PMID- 2543958 TI - The nucleotide sequence of the lacC and lacD genes of Staphylococcus aureus. PMID- 2543959 TI - Sequence variation of the Rous sarcoma virus PrA src gene. PMID- 2543960 TI - Complete nucleotide sequence of an E.coli IS5 insertion element containing an internal 88 base pair direct repeat (IS5-D). PMID- 2543961 TI - Sequence of the cDNA for the alpha subunit of calmodulin kinase II from mouse brain. PMID- 2543962 TI - DNA sequence analysis of the inverted terminal repeats of a non-oncogenic avian adenovirus. PMID- 2543963 TI - An improved retroviral vector for gene transfer into undifferentiated cells. PMID- 2543964 TI - Estradiol-progesterone-induced growth of the rat dorsolateral prostate: effects on protein phosphorylation. AB - The combination of estradiol and progesterone can affect growth of the rat dorsolateral prostate by a mechanism that appears to be independent from androgen mediated activity. The effects of estradiol and progesterone on protein kinase activity and endogenous protein phosphorylation in the castrate rat dorsolateral prostate were compared with androgenic effects on these intracellular events. Differences in protein kinase activity and protein phosphorylation were found with each type of hormonal therapy. Changes in protein kinase activity and protein phosphorylation produced by estradiol and progesterone in the rat dorsolateral prostate were not found in the rat ventral prostate, which was not affected by these hormones in the 28 day length of this study. PMID- 2543965 TI - Changes in placental atrial natriuretic peptide receptors associated with severe toxemia of pregnancy. AB - We have previously demonstrated the presence of atrial natriuretic peptide (ANP) specific receptors in normal human placentas. Since toxemia of pregnancy may affect important metabolic, transport and hemodynamic functions of placentas, we have asked the question whether binding properties of human placental ANP receptors are changed in patients with severe toxemia of pregnancy when compared to normal patients. ANP receptors in plasma membranes from normal and severely toxemic patients were characterized by in vitro radioligand assays utilizing [125I]-alpha-hANP. In all cases, we identified specific, high affinity, low capacity ANP binding sites in a microsomal fraction of human placentas. Although the total concentration of receptors did not differ between the two groups, the dissociation constant, KD, was significantly higher in placentas from severely toxemic patients than from normal controls. From the above we conclude that placental ANP receptors are dynamically modulated and their characteristics may be altered in severe toxemia of pregnancy. PMID- 2543966 TI - Uptake of bicarbonate/CO2 in the isolated guinea pig placenta. AB - Experiments were performed to investigate the mechanisms of bicarbonate/CO2 transport in the isolated guinea pig placenta. The maximal uptake of [14C] bicarbonate/CO2 (reference [3H]-L-glucose) was about 60 per cent on both the fetal and maternal side, using the method of paired tracer single injection dilution technique. Carbonic anhydrase inhibitors such as acetazolamide (0.1 and 1 mmol/l) and the poorly permeant aminothiadiazole (0.5 mmol/l) reduced bicarbonate/CO2 uptake on both sides by 25-50 per cent. Inhibition was significantly higher on the fetal side. When lactate (5 mmol/l) was added to the acceptor side solution, the uptake of bicarbonate/CO2 increased 10-20 per cent. DIDS (1 mmol/l) decreased bicarbonate/CO2 uptake additionally under the simultaneous influence of acetazolamide. It is concluded that at least two mechanisms for placental bicarbonate transfer exist. A portion of bicarbonate, predominantly on the fetal side and outside the trophoblastic cells, is converted to CO2 which then diffuses into the tissue and crosses the placental barrier. Another part of bicarbonate is transported as ion using an anion exchange system, which exists in both the multivillous and the basal membranes of the syncytium. The first system is important for the excretion of fetal CO2 into the mother. PMID- 2543967 TI - Massive hemorrhage as the initial appearance of "nonfunctioning" islet cell tumors of the pancreas. AB - "Nonfunctioning" tumors of the endocrine pancreas usually appear at a late stage with abdominal mass or jaundice. We describe two patients with nonfunctioning islet cell tumors who had the rare complication of massive hemorrhage. In each case, the hemorrhage was successfully controlled by an operation. PMID- 2543968 TI - Identification of protease(s) capable of further degrading native 3/4- and 1/4 collagen fragments generated by collagenase from native type I collagen in human neutrophils. AB - The role of human neutrophil proteases in the further degradation of the native triple-helical characteristic cleavage products 3/4- and 1/4-collagen fragments generated by neutrophil interstitial collagenase from native type I collagen was studied. Purified human neutrophil collagenase did not further degrade the characteristic collagen fragments whether they were in triple-helical (native collagen) or random-coil (gelatin) conformation. Neutrophil extract treated with 1 mM phenylmercuric chloride (PMC) degraded native type I collagen at +37 degrees C producing multiple protein bands. Neutrophil extract at +18 degrees C in the presence of the serine protease inhibitors phenylmethylsulfonyl fluoride and banzamidine did not degrade native type I collagen. Inclusion of PMC to active latent collagenase caused neutrophil extract to degrade native type I collagen to 3/4- and 1/4-fragments. In addition, native 3/4- and 1/4-fragments were further degraded in a time-dependent manner by PMC-treated neutrophil extract. Both native 3/4- and 1/4-collagen fragments were degraded by specific rather than by multiple cleavage. Further fragmentation was inhibited by divalent cation chelators EDTA and 1,10-phenanthroline. The results indicate the presence of latent metalloprotease(s), as distinct from collagenase, gelatinase and serine proteases, that are capable of further degrading by specific cleavage both native 3/4- and 1/4-collagen fragments generated by collagenase in human neutrophils. The enzyme(s) may augment the action of collagenase and other neutral proteases in connective tissue destruction associated with the etiopathogenesis of periodontal diseases. PMID- 2543969 TI - Correlation of paramagnetic states and molecular structure in bacterial photosynthetic reaction centers: the symmetry of the primary electron donor in Rhodopseudomonas viridis and Rhodobacter sphaeroides R-26. AB - The orientation of the principal axes of the primary electron donor triplet state measured in single crystals of photosynthetic reaction centers is compared to the x-ray structures of the bacteria Rhodobacter (Rb.) sphaeroides R-26 and Rhodopseudomonas (Rps.) viridis. The primary donor of Rps. viridis is significantly different from that of Rb. sphaeroides. The measured directions of the axes indicate that triplet excitation is almost completely localized on the L subunit half of the dimer in Rps. viridis but is more symmetrically distributed (approximately 63% on the L half of the special pair and approximately 37% on the M half) on the dimeric donor in Rb. sphaeroides R-26. The large reduction of the zero field splitting parameters relative to monomeric bacteriochlorophyll triplet in vitro suggests significant participation of asymmetrical charge transfer electronic configurations in the special pair triplet state of both organisms (approximately 23% in Rps. viridis and approximately 13% in Rb. sphaeroides). PMID- 2543970 TI - Import of cytochrome c into mitochondria: reduction of heme, mediated by NADH and flavin nucleotides, is obligatory for its covalent linkage to apocytochrome c. AB - The covalent attachment of heme to apocytochrome c, and therefore the import of cytochrome c into mitochondria, is dependent on both NADH plus a cytosolic cofactor that has been identified to be FMN or FAD. NADH in concert with flavin nucleotides mediates the reduction of heme. Heme in the reduced state is a prerequisite for its covalent attachment to apocytochrome c by the enzyme cytochrome c heme lyase and thus for subsequent translocation of cytochrome c across the outer mitochondrial membrane during import. PMID- 2543971 TI - Activation of human CDC2 protein as a histone H1 kinase is associated with complex formation with the p62 subunit. AB - p34 kinase, the product of the CDC2 gene, is a cell-cycle regulated protein kinase that is most active during mitosis. In HeLa cells, p34 kinase has previously been shown to exist in both a low- and a high-molecular-mass form, the latter of which is only found in cells in the G2/M phase of the cell cycle and contains a 62-kDa subunit. Here we show that although each form of the kinase phosphorylates casein in vitro, only the high-molecular-mass form uses histone H1 as substrate. The high-molecular-mass form of p34 kinase from nocodazole-treated HeLa cells was purified 6700-fold. The apparent molecular mass of the mitotic CDC2-encoded protein kinase complex was 220 kDa. The purified enzyme phosphorylated not only its endogenous 62-kDa subunit but also phosphorylated histone H1 with a Km of 3 microM and used ATP 40 times more efficiently than GTP (Km 54 microM and 2 mM, respectively). The enzyme activity was unaffected by cAMP, calcium/calmodulin, or by the heat-stable inhibitor of cAMP-dependent protein kinase. These characteristics are typical of growth-associated histone H1 kinase from different organisms. These results suggest that CDC2 protein may be activated as an M-phase-specific protein kinase in part by its association with the p62 subunit. PMID- 2543972 TI - Dual affinity fusion approach and its use to express recombinant human insulin like growth factor II. AB - A dual affinity fusion concept has been developed in which the gene encoding the desired product is fused between two flanking heterologous genes encoding IgG- and albumin-binding domains. Using sequential IgG and serum albumin affinity chromatography, a full-length tripartite fusion protein is obtained. This approach was used to recover a full-length fusion product in Escherichia coli containing the human insulin-like growth factor II (IGF-II). Surprisingly, the recombinant IGF-II showed increased stability against proteolytic degradation in E. coli when produced as a dual affinity fusion protein, as compared to an N terminal fusion protein. After site-specific cleavage of the tripartite fusion protein, IGF-II molecules with immunological and receptor binding activity were obtained without renaturation steps. The results demonstrate that proteins can fold into biologically active structures, even if provided with large flanking heterologous protein domains. The concept was further used to characterize the specific degradation of recombinant IGF-II in this heterologous host. PMID- 2543973 TI - Centrally truncated and stabilized porcine neuropeptide Y analogs: design, synthesis, and mouse brain receptor binding. AB - Porcine neuropeptide Y (pNPY) has been proposed to form an intramolecularly stabilized structure characterized by N- and C-terminal helical regions arranged antiparallel due to a central turn region. Analogs based on this structural model that have the central turn region and various amounts of the helical regions removed, yet retain the N and C termini in a similar spatial orientation were designed. The gap formed by removal of the central residues (residues 8-17 or 7 20) was spanned with a single 8-aminooctanoic acid residue (Aoc) and the structure was further stabilized by the introduction of a disulfide bridge. [D Cys7,Aoc8-17,Cys20]pNPY and [Cys5,Aoc7-20,D-Cys24]pNPY were synthesized and found to have receptor binding affinities of 2.3 nM and 150 nM, respectively, in mouse brain membranes (pNPY affinity is 3.6 nM in this assay). It is proposed that the central region (residues 7-17) of pNPY serves a structural role in the peptide and is not involved in direct receptor interaction. PMID- 2543975 TI - Molecular cloning of the alpha-subunit of human prolyl 4-hydroxylase: the complete cDNA-derived amino acid sequence and evidence for alternative splicing of RNA transcripts. AB - Prolyl 4-hydroxylase [procollagen-proline, 2-oxyglutarate 4-dioxygenase; procollagen-L-proline, 2-oxoglutarate:oxygen oxidoreductase (4-hydroxylating), EC 1.14.11.2], an alpha 2 beta 2 tetramer, catalyzes the formation of 4 hydroxyproline in collagens by the hydroxylation of proline residues in peptide linkages. We report here on the isolation of cDNA clones encoding the alpha subunit of the enzyme from human tumor HT-1080, placenta, and fibroblast cDNA libraries. Eight overlapping clones covering almost all of the corresponding 3000 nucleotide mRNA, including all the coding sequences, were characterized. These clones encode a polypeptide of 517 amino acid residues and a signal peptide of 17 amino acids. Previous characterization of cDNA clones for the beta-subunit of prolyl 4-hydroxylase has indicated that its C terminus has the amino acid sequence Lys-Asp-Glu-Leu, which, it has been suggested, is necessary for the retention of a polypeptide within the lumen of the endoplasmic reticulum. The alpha-subunit does not have this C-terminal sequence, and thus one function of the beta-subunit in the prolyl 4-hydroxylase tetramer appears to be to retain the enzyme within this cell organelle. Interestingly, three of the cDNA clones for the alpha-subunit contained a 64-nucleotide sequence homologous but not identical to the corresponding 64-nucleotide sequence found in four other cDNA clones. Nuclease S1 mapping experiments demonstrated that this difference was due to the existence of two types of mRNA present in approximately equal amounts. Southern blot analyses of human genomic DNA with a cDNA probe for the alpha-subunit suggested the presence of only one gene encoding the two types of mRNA, which appear to result from mutually exclusive alternative splicing of primary transcripts of one gene. PMID- 2543974 TI - Mammalian aspartate transcarbamylase (ATCase): sequence of the ATCase domain and interdomain linker in the CAD multifunctional polypeptide and properties of the isolated domain. AB - Mammalian aspartate transcarbamylase (ATCase; carbamoyl-phosphate:L-aspartate carbamoyltransferase, EC 2.1.3.2) is part of a 240-kDa multifunctional polypeptide called CAD, which also has carbamoyl-phosphate synthetase and dihydroorotase activities. We have sequenced selected restriction fragments of a Syrian hamster CAD cDNA that are clearly homologous to three prokaryotic ATCases. These studies, combined with previous sequence data, showed that the ATCase domain of CAD is encoded by 924 base pairs and has a mass of 34,323 Da and a pI of 9.8. While the bacterial pyrimidine biosynthetic enzymes are separate proteins, in mammals the ATCase domain is fused to the carboxyl end of the CAD chimera via a 133-amino acid (14-kDa) linker with an unusual amino acid composition, a pI of 10.2, and pronounced hydrophilic character. The fully active domain isolated from proteolytic digests was characterized by partial amino acid sequencing and amino acid analysis. Trypsin cleavage produced the ATCase domain with a 20-residue amino-terminal extension. Hydrodynamic studies showed that the isolated domain is a 110-kDa trimer with a Stokes radius of 41 A. The mammalian ATCase domain and the prokaryotic enzymes have virtually identical active-site residues and are likely to have the same tertiary fold. PMID- 2543976 TI - Methionine synthesis in Escherichia coli: effect of the MetR protein on metE and metH expression. AB - Studies by Urbanowski et al. [Urbanowski, M. L., Stauffer, L. T., Plamann, L. S. & Stauffer, G. V. (1987) J. Bacteriol. 169, 1391-1397] have identified a regulatory locus, called metR, required for the expression of the metE and metH genes. We recently purified the MetR protein from Escherichia coli and showed that it could stimulate the in vitro expression of the metE gene and autoregulate its own synthesis. In the present study, the purified MetR protein has been shown to stimulate the in vitro expression of the metH gene. Also, the in vitro synthesized MetE, MetH, and MetR proteins were shown to be biologically active. The transcription start sites for the metE and metR genes have been determined, and DNA footprinting experiments have identified regions in the metE-metR intergenic sequence that are protected by either the MetR or MetJ proteins. PMID- 2543977 TI - Escherichia coli DNA helicase II (uvrD gene product) catalyzes the unwinding of DNA.RNA hybrids in vitro. AB - DNA helicase II is a well-characterized Escherichia coli enzyme capable of unwinding duplex DNA and known to be involved in both methyl-directed mismatch repair and excision repair of pyrimidine dimers. Here it is shown that this enzyme also catalyzes the ATP-dependent unwinding of a DNA.RNA hybrid consisting of a radioactively labeled RNA molecule annealed on M13 single-stranded DNA. The DNA.RNA unwinding reaction required less protein to unwind more base pairs than the corresponding unwinding of duplex DNA. In addition, the rate of unwinding of the DNA.RNA hybrid was more than an order of magnitude faster than unwinding of a DNA partial duplex of similar length. The unwinding of the DNA.RNA hybrid is a property unique to helicase II since helicase I, Rep protein, and helicase IV failed to catalyze the reaction. In light of these results it seems likely that helicase II is involved in some previously unrecognized aspect of nucleic acid metabolism, in addition to its known roles in DNA repair reactions. PMID- 2543978 TI - Oxygen permeability of phosphatidylcholine--cholesterol membranes. AB - Oxygen transport in phosphatidylcholine-cholesterol membranes has been studied by observing the collision of molecular oxygen with nitroxide radical spin labels placed at various distances from the membrane surface using long-pulse saturation recovery ESR techniques. The collision rate was estimated for tempocholine phosphatidic acid ester, 5-doxylstearic acid, and 16-doxylstearic acid from spin lattice relaxation times (T1) measured in the presence and absence of molecular oxygen. Profiles of the local oxygen transport parameter across the membrane were obtained as a function of cholesterol mol fraction and temperature in L-alpha dimyristoylphosphatidylcholine ([ Myr2]PtdCho) and L-alpha dioleoylphosphatidylcholine ([ Ole2]PtdCho) membranes. Membrane oxygen permeability coefficients were estimated from oxygen transport parameter profiles. At approximately 30 degrees C, the oxygen permeability coefficients in the presence and absence of 50 mol % cholesterol are 22.7 and 125.2 cm/s, respectively, for [Myr2]PtdCho membranes, and 54.7 and 114.2 cm/s, respectively, for [Ole2]PtdCho membranes (compared with 60-80 cm/s for water layers with the same thicknesses as the membranes). The major results in the liquid-crystalline phase are as follows: (i) In the absence of cholesterol, membranes are not barriers to oxygen transport. (ii) Addition of 50 mol % cholesterol decreases oxygen permeability by a factor of approximately 5 and approximately 2.5 in [Myr2]PtdCho and [Ole2]PtdCho membranes, respectively. The resistance to oxygen transport is located in and near the polar headgroup regions in the membrane. (iii) Cholesterol increases oxygen transport in the central regions of [Ole2]PtdCho membranes. PMID- 2543979 TI - Molecular properties of pyruvate bound to lactate dehydrogenase: a Raman spectroscopic study. AB - Lactate dehydrogenase (LDH; EC 1.1.1.27) catalyzes the addition of pyruvate to the four position of the nicotinamide ring of bound NAD+; this NAD-pyruvate adduct is bound tightly to the enzyme. We have used the adduct as a model for pyruvate in a competent ternary complex by comparing the Raman spectrum of the bound adduct with that for unliganded pyruvate. To understand the observed normal modes of pyruvate both as the bound adduct and in water, we have taken the Raman spectra of a series of 13C- and 18O-labeled pyruvates. We find that the carboxylate COO- moiety of pyruvate remains unprotonated at LDH's active site and forms an ion pair complex. The frequency of pyruvate's carbonyl C = O moiety shifts from 1710 cm-1 in water downward 34 cm-1 when pyruvate binds to LDH. This frequency shift corresponds to a ca. 34% polarization of the carbonyl bond, indicates a substantial interaction between the C = O group and enzyme, and is direct evidence for and is a measure of enzyme-induced electronic perturbation of the substrate needed for catalysis. This bond polarization is likely brought about by electrostatic interactions between the carbonyl moiety and the protonated imidazole group of His-195 and the guanidino group from Arg-109. We discuss how the data bear on the enzymatic chemistry of LDH. PMID- 2543980 TI - Failure of glucose to elicit a normal secretory response in fetal pancreatic beta cells results from glucose insensitivity of the ATP-regulated K+ channels. AB - Fetal pancreatic beta cells demonstrate a deficient insulin release in response to glucose, but the underlying mechanism at the cellular level is unknown. By using beta cells from 21-day fetal rats we made an attempt to clarify the mechanism(s) behind this reduced glucose response. In addition to measuring insulin release, glucose metabolism, and cellular ATP content, ATP-regulated K+ channels (G channels) and voltage-activated Ca2+ currents were investigated with the patch-clamp technique. It was thus demonstrated that the ATP-regulated K+ channels in fetal beta cells were not sensitive to glucose but otherwise had similar characteristics as those of adult beta cells. Also, the characteristics of the voltage-activated Ca2+ currents were similar in adult and fetal beta cells. However, as judged from measurements of both glucose oxidation and glucose utilization, glucose metabolism was impaired in fetal beta cells. In addition, there was no increase in the ATP content, even when the cells were stimulated for 30 min. It is therefore concluded that the attenuated glucose-induced insulin release in fetal pancreatic beta cells is due to an immature glucose metabolism resulting in impaired regulation of the ATP-sensitive K+ channels. These findings may be relevant to the understanding of the deficient stimulus-secretion coupling associated with non-insulin-dependent diabetes. PMID- 2543981 TI - Induction of cervical neoplasia in the mouse by herpes simplex virus type 2 DNA. AB - Induction of cervical neoplasia in the mouse cervix by herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) has been reported. The present study was done to determine if transfection with DNA of HSV-2 can induce carcinogenesis in this animal model. Genomic HSV-2 DNA was isolated from infected HEp-2 cells and separated from host cell DNA by cesium chloride density gradient centrifugation. The DNA was applied to mouse cervix for periods of 80-100 weeks. Experimental controls were treated with uninfected genomic HEp-2 cell DNA or with calf thymus DNA. Vaginal cytological preparations from all animals were examined monthly to detect epithelial abnormalities. Animals were sacrificed and histopathology studies were done when cellular changes indicative of premalignant or malignant lesions were seen on vaginal smears. Cytologic and histologic materials were coded and evaluated without knowledge of whether they were from animals treated with virus or control DNA. Premalignant and malignant cervical lesions similar to those that occur in women were detected in 61% of the histologic specimens obtained from animals exposed to HSV-2 DNA. The yield of invasive cancers was 21% in animals treated with HSV-2 DNA. No cancers were detected in mice treated with either HEp-2 or calf thymus DNA. Dysplasia was detected in only one of these control animals. PMID- 2543982 TI - Growth- and tumor-promoting effects of deregulated BCL2 in human B-lymphoblastoid cells. AB - Human follicular B-cell lymphomas possess a t(14;18) that translocates a putative protooncogene, BCL2, into the immunoglobulin heavy chain locus. The normal BCL2 gene is quiescent in resting B cells, expressed in proliferating, but down regulated in differentiated B cells. Inappropriately high levels of BCL2 immunoglobulin chimeric RNA are present in t(14;18) lymphomas for their mature B cell stage. We examined the biologic effects of BCL2 deregulation in human B cells by introducing BCL2 into human B-lymphoblastoid cell lines (LCLs) with retroviral gene transfer. Although deregulated BCL2 expression as a single agent was not sufficient to confer tumorigenicity to LCLs, it consistently produced a 3 to 4-fold increment in LCL clonogenicity in soft agar. In addition, BCL2 deregulation complements the transforming effects of the MYC oncogene in LCLs. BCL2 augmented the clonogenicity of LCLs bearing exogenous MYC and increased the frequency and shortened the latency of tumor induction in immunodeficient mice. These results demonstrate a role for BCL2 as a protooncogene that affects B-cell growth and enhances B-cell neoplasia. PMID- 2543983 TI - Frameshifting is required for production of the transposase encoded by insertion sequence 1. AB - Insertion sequence IS1 has two coding frames, insA and insB, which are essential for its transposition. Here, we show that a frameshifting event in the -1 direction from the 3' end region of the insA frame to an open reading frame (B' frame), extending from the 5' end of the insB frame, is involved in production of the InsA-B'-InsB fusion protein that has IS1 transposase activity. The frameshifting event is likely to have occurred at the sequence AAAAAC where the insA frame overlaps the B' frame. Interestingly, this sequence is also present in one of the two sequences identified in retroviruses as frameshift signals for production of transframe polyproteins from the overlapping genes gag-pro or gag pro-pol. PMID- 2543984 TI - Direct evidence that oxygen-derived free radicals contribute to postischemic myocardial dysfunction in the intact dog. AB - Electron paramagnetic resonance (EPR) spectroscopy was used to investigate whether (i) the free radicals produced in the "stunned" myocardium (myocardium with postischemic contractile dysfunction) are derived from O2, (ii) inhibition of radical reactions improves function, and (iii) i.v. spin traps are effective. Open-chest dogs undergoing a 15-min coronary occlusion received an i.v. infusion of the spin trap, alpha-phenyl N-tert-butylnitrone (PBN) (50 mg/kg). In group I (n = 6), EPR signals characteristic of radical adducts of PBN appeared in the coronary venous blood during ischemia and increased dramatically after reperfusion. In group II (n = 6), which received PBN and i.v. superoxide dismutase (SOD; 16,000 units/kg) plus catalase (12,000 units/kg), myocardial production of PBN adducts was undetectable during ischemia (delta = -100%, P less than 0.01 vs. group I) and markedly inhibited after reperfusion (delta = -86%, P less than 0.001). This effect was seen at all levels of ischemic zone flow but was relatively greater in the low-flow range. In group III (n = 8), the same dosages of SOD and catalase without PBN markedly enhanced contractile recovery (measured as systolic wall thickening) after reperfusion [P less than 0.01 at 3 hr vs. controls (group IV, n = 7)]. Systemic plasma activity of SOD and catalase averaged 127 +/- 24 and 123 +/- 82 units/ml, respectively, 2 min after reperfusion. PBN produced no apparent adverse effects and actually improved postischemic contractile recovery in group I (P less than 0.05 at 3 hr vs. controls). This study shows that (i) SOD and catalase are highly effective in blocking free radical reactions in vivo, (ii) the radicals generated in the "stunned" myocardium are derived from univalent reduction of O2, and (iii) inhibition of radical reactions improves functional recovery. The results provide direct, in vivo evidence to support the hypothesis that reactive oxygen metabolites play a causal role in the myocardial "stunning" seen after brief ischemia. PMID- 2543991 TI - Conditioned stereotypy: behavioral specification of the UCS and pharmacological investigation of the neural change. AB - Previous work has shown that conditioned stereotypy can be produced by repeated treatments with d-amphetamine or apomorphine. We replicated this phenomenon and found that, as in previous reports, the amplitude of conditioned stereotypy was about one-third that of the unconditioned stereotypy. On the basis of the hypothesis that the UCS in this conditioning situation is a specific stimulation level of dopamine receptors expressed as a peak behavioral effect (UCR), rats were exposed to the experimental boxes for a brief interval during the peak behavioral effect of the drugs. This procedure produced an amplitude of conditioned stereotypy about two-thirds that of unconditioned stereotypy. The issue of the synaptic mechanism mediating conditioned stereotypy was addressed by examining the effect of pimozide on the behavior. A dose of pimozide that completely blocked apomorphine-unconditioned stereotypy also blocked apomorphine conditioned stereotypy with no sign of motor impairment. d-Amphetamine conditioned stereotypy was not completely blocked by a dose of pimozide that completely blocked d-amphetamine-unconditioned stereotypy. The implications of these findings for understanding the neural basis of conditioned stereotypy are discussed. PMID- 2544001 TI - Biochemical and functional aspects on the control of prolactin release by the hypothalamo-pituitary GABAergic system. AB - A growing body of biochemical, immunohistochemical, and autoradiographic evidence indicates the presence of two different GABAergic systems in the mediobasal hypothalamus: one intrinsic, the tuberoinfundibular GABAergic system, and the other extrinsic, whose cell bodies are located outside the mediobasal hypothalamus and which projects to this area and establishes synaptic contacts with aminergic and peptidergic neurons involved in endocrine function. This particular anatomical configuration provides a rational basis to explain the dual action of GABA (inhibitory and stimulatory) on prolactin release. Different studies aimed at identifying the precise role of GABA on prolactin function have demonstrated that this system can be modulated, at the pre- and/or post-synaptic level, by different experimental maneuvers in which prolactin secretion is physiologically and pharmacologically altered. GABA mainly appears to be involved in feedback mechanisms preventing an exaggerated prolactin output during specific physiological situations. The ability of clinically tested, direct GABAmimetic compounds to lower prolactin secretion in the rat points towards a clinical usefulness of these drugs in particular spontaneous or induced neuroendocrine disorders. However, the possibility of a widespread use of this type of compounds is hampered by the lack of potent, specific and non-toxic GABA agonists suitable for clinical purposes. PMID- 2544002 TI - The assessment of abnormalities in hormonal responsiveness at multiple levels of the hypothalamic-pituitary-adrenocortical axis in depressive illness. AB - A substantial body of data suggests that excessive cortisol secretion in depression may result from dysregulation at several sites within the hypothalamic pituitary-adrenocortical (HPA) axis. The alterations in regulatory mechanisms are thought to result from a limbic system-hypothalamic "overdrive" of corticotropin releasing hormone (CRH). We also have demonstrated that excessive secretion of cortisol may result from an abnormal adrenocortical responsiveness to adrenocorticotropic hormone (ACTH), and we have postulated that corticotropic cells within the pituitary mediate between excessive secretion of CRH from the hypothalamus and hypercortisolemia secondary to adrenocortical hyperplasia and enhanced sensitivity to ACTH at the adrenal cortex. The present report describes a series of clinical experiments utilizing several neuroendocrine probes, as well as computer-assisted tomography, to examine the complexities of the HPA axis dysregulation in depression. These studies support the hypothesis that a limbic system-hypothalamic disturbance results in excessive CRH secretion as well as enhanced adrenocortical activity, and that these factors contribute to excessive cortisol secretion in patients with depression. These data further support the hypothesis that endogenous affective disorders are best characterized in the framework of a generalized biological disturbance of HPA axis function which involves both central and peripheral endocrine sites. PMID- 2544004 TI - Transposable elements in mutagenesis and regulation of gene expression. PMID- 2544005 TI - Molecular lesions induced by I-R hybrid dysgenesis in Drosophila melanogaster. PMID- 2544007 TI - Transposition of intracisternal A-particle genes. PMID- 2543995 TI - An ESR study of the visible light photochemistry of gilvocarcin V. AB - Photolysis of gilvocarcin (GV) at 405 nm in argon saturated dimethylsulfoxide (DMSO) or 50% DMSO-water solutions in the presence of the sodium salt of 3,5 dibromo-2,6-dideutero-4-nitrosobenzene sulfonic acid (DBNBS-d2) generates the CH3 DBNBS-d2.spin adduct. It is postulated that this spin adduct is produced by photoreduction of DMSO by GV and the consequent formation and trapping of the generated methyl radicals. Gilvocarcin V also photoreduces oxygen and methyl viologen with quantum yields of 0.019 and 0.0012 respectively. The quantum yield for singlet oxygen formation by GV in DMSO, determined by measuring the rate of production of the nitroxyl radical produced by the reaction of 2,2,6,6 tetramethylpiperidinol with singlet oxygen, was found to be 0.15. Thus, GV photochemistry proceeds by both Type I and Type II pathways which could contribute to the reported GV phototoxicity in biological systems. PMID- 2544006 TI - Insertional mutations in transgenic mice. AB - Insertional mutagenesis represents a promising approach to the identification of new genes involved in mammalian development. In this paper, we have presented a brief review of the literature on the analysis of mutations caused by DNA and retroviral insertion into the mouse genome. We have discussed several methods that we and others have used to identify recessive insertional mutations among transgenic mouse lines. Finally, we have summarized the results of our studies to date on three recessive prenatal lethal mutations that we have identified. PMID- 2543996 TI - Endorphins in male impotence: evidence for naltrexone stimulation of erectile activity in patient therapy. AB - In the present study we evaluated whether naltrexone administration could stimulate sexual function in 30 male patients, ages 25 to 50 years, with idiopathic impotence of at least one year's duration and not of organic etiology. The patients received naltrexone (50 mg/day) or placebo, on a random basis for two weeks. Sexual performance, expressed as the number of full coitus/week, was assessed before (time 0) and during (on days 7 and 15) each treatment. The naltrexone therapy significantly increased the number of successful coitus compared to placebo after 7 and 15 days of treatment: improvement of sexual performance was evident in 11 out of the 15 treated patients. All the patients experienced a significant increase in morning and spontaneous full penile erections/week. No significant side effects were reported. Endocrine studies revealed no significant modification of plasma LH, FSH or testosterone by naltrexone, suggesting that the positive effect of the drug on sexual behavior was exerted at a central level. A two-month follow-up, at which time patients were off treatment, erectile capacity had returned to baseline in 10 patients, while five reported complete recovery of their sexual ability. We hypothesize that an alteration in central opioid tone is present in idiopathic impotence and is involved in the impairment of sexual behavior. PMID- 2543998 TI - Evidence for a functional role of alpha-1 adrenoceptors in the regulation of melatonin secretion in man. AB - Nocturnal plasma melatonin concentrations were reduced following administration of the alpha 1 adrenoceptor antagonist, prazosin (2 mg orally at 1900h), to 10 normal men. These data indicate that alpha 1 adrenoceptors are involved in the physiological control of melatonin secretion in man. PMID- 2543992 TI - Effects of GABA antagonists on the pentobarbital-induced depression of respiration and cough in rats. AB - In order to determine the possible involvement of GABA-ergic mechanisms in the modulation of the cough reflex, the effects of GABA antagonists on the pentobarbital-induced depression of respiration and cough were examined in a comparative study in rats. The cough reflex was induced by application of electrical stimulation to the tracheal mucosa by the puncture electrode-induced cough method. The 50% antitussive dose (AtD50) of pentobarbital was calculated by the "up and down" method. Pentobarbital (10 mg/kg, IP) caused a reduction of tidal volume, which was counteracted by pretreatment with picrotoxin (3 mg/kg, IP) or bicuculline (3 mg/kg, IP). However, neither picrotoxin nor bicuculline were able to counteract the reduction in frequency of respiration. The AtD50 of pentobarbital was 1.95 mg/kg when administered IP. The AtD50 of pentobarbital was not altered after pretreatment of rats with picrotoxin (1.85 mg/kg, IP) or with bicuculline (1.55 mg/kg). These results suggest that GABA-ergic mechanisms may not be involved in the cough-depressant effect of pentobarbital. PMID- 2544008 TI - A retroviral insertion in the dilute (d) locus provides molecular access to this region of mouse chromosome 9. PMID- 2544009 TI - Spontaneous germ-line ecotropic murine leukemia virus infection: implications for retroviral insertional mutagenesis and germ-line gene transfer. PMID- 2543985 TI - Thymidine kinase-negative herpes simplex virus mutants establish latency in mouse trigeminal ganglia but do not reactivate. AB - Herpes simplex virus infection of mammalian hosts involves lytic replication at a primary site, such as the cornea, translocation by axonal transport to sensory ganglia and replication, and latent infection at a secondary site, ganglionic neurons. The virus-encoded thymidine kinase, which is a target for antiviral drugs such as acyclovir, is not essential for lytic replication yet evidently is required at the secondary site for replication and some phase of latent infection. To determine the specific stage in viral pathogenesis at which this enzyme is required, we constructed virus deletion mutants that were acyclovir resistant and exhibited no detectable thymidine kinase activity. After corneal inoculation of mice, the mutants replicated to high titers in the eye but were severely impaired for acute replication in trigeminal ganglia and failed to reactivate from ganglia upon cocultivation with permissive cells. Nevertheless, latency-associated transcripts were expressed in neuronal nuclei of ganglia from mutant-infected mice and superinfection of the ganglia with a second virus rescued the latent mutant virus. Thus, contrary to a widely accepted hypothesis, the thymidine kinase-negative mutants established latent infections, implying that neither thymidine kinase activity nor ganglionic replication is necessary for establishment of latency. Rather, thymidine kinase appears to be necessary for reactivation from latency. These results suggest that acyclovir-resistant viruses could establish latent infections in clinical settings and have implications for the use of genetically engineered herpesviruses to deliver foreign genes to neurons. PMID- 2544011 TI - Transposable elements in natural populations of Drosophila. PMID- 2544010 TI - Mouse endogenous retroviral long-terminal-repeat (LTR) elements and environmental carcinogenesis. PMID- 2544012 TI - Drosophila foldback elements, primate L1 elements, and transgenic mice. PMID- 2544003 TI - Neurophysiological and endocrine consequences of immune activity. AB - The studies presented herein demonstrate the potency with which activity of the immune system is able to influence the central nervous system. Electrophysiological recordings have demonstrated significant changes in preoptic area/anterior hypothalamic (PO/AH) multiunit electrical activity (MUA) following sensitization with sheep red blood cells. The peak of activity occurred on the fifth day after immunization, the same day that serum antibodies were first detected. A significant increase in paraventricular nucleus MUA was also demonstrated, but this appeared to be delayed with respect to that in the PO/AH, occurring on the sixth day. Further changes thought to be associated with the immune response also were found: Serum corticosterone levels were elevated on the eighth day of the response, and PO/AH tissue levels of norepinephrine were reduced between the sixth and tenth days. During induction of a secondary response, PO/AH MUA showed a different profile of activity from that recorded during the first response. Chronic administration of the immunosuppressive drug, cyclophosphamide, prevented the recorded changes in PO/AH MUA. These results suggest that some secretory product(s) of the activated immune system may be able to exert effects on the central nervous system. Various immunoactive substances therefore were administered intra-cerebroventricularly in order to examine their effects upon PO/AH MUA, cortical EEG and adrenocortical hormone secretory activity. alpha-Interferon and thymic humoral factor were both found to decrease PO/AH MUA, increase EEG synchronization, and decrease basal levels of circulating corticosterone. In contrast, histamine and interleukin-1 did not alter PO/AH MUA but did cause decreased EEG synchronization and increased serum corticosterone levels. With another preparation, a specific activating effect of interleukin-1 upon putative corticotropin-releasing factor-secreting neurones has also been found, identified vasopressinergic neurones not being affected. PMID- 2544013 TI - The hobo element of Drosophila melanogaster. PMID- 2544014 TI - Molecular biology of Drosophila P-element transposition. PMID- 2544016 TI - Spread of P transposable elements in inbred lines of Drosophila melanogaster. PMID- 2544017 TI - Suppressible insertion-induced mutations in Drosophila. PMID- 2543999 TI - Effect of naloxone on the growth hormone response to clonidine in normal women during the mid-luteal phase. AB - We studied the effect of the opiate antagonist naloxone on the peripheral GH response to the alpha 2-receptor agonist clonidine in eight normally cycling women during the mid-luteal phase. In a randomized, double-blind, cross-over design, each subject received clonidine and naloxone on one occasion and clonidine and placebo on the other. In seven of eight subjects, an attenuation of the GH response was associated with naloxone administration. The maximal GH increment above baseline (delta GHMAX) of 7.8 +/- 2.0 micrograms/L (mean +/- SEM) with placebo was higher than the delta GHMAX of 4.2 +/- 0.9 micrograms/L with naloxone (p = 0.05). Likewise, the area above baseline under the GH level-time curve following clonidine (delta GHAREA) was higher with placebo compared to naloxone (477 +/- 175 micrograms/L x min vs. 228 +/- 62 micrograms/L x min), although this difference was not quite statistically significant (p = 0.09). As expected, with placebo the increase in GH following clonidine was statistically significant by repeated measures analysis of variance (p = 0.001). The smaller increase in GH levels when naloxone was given was not significant. Both delta GHMAX and delta GHAREA values were significantly positively correlated with estradiol levels when placebo was given, but not when naloxone was given. GHRH was not detectable following clonidine administration under either the placebo or the naloxone conditions. Our data support the hypothesis that estrogen enhances the response of GH to provocative stimuli in women, at least in part by increasing endogenous opioid tone in the hypothalamus. PMID- 2544018 TI - Identifying and cloning Drosophila genes by single P element insertional mutagenesis. PMID- 2543994 TI - Antiviral effects of phosphonoformate (PFA, foscarnet sodium). AB - This article describes the antiviral properties of foscarnet (trisodium phosphonoformate) at the enzyme level as well as in cell cultures and in vivo. The mechanism of action against herpesvirus DNA polymerases and reverse transcriptases is outlined. Clinical studies using topical foscarnet against mucocutaneous herpes simplex virus infections are presented. The clinical use of intravenous foscarnet against severe viral infections caused by cytomegalovirus, hepatitis B virus and human immunodeficiency virus is discussed. PMID- 2544000 TI - Helpless behavior (escape deficits) in streptozotocin-diabetic rats: resistance to antidepressant drugs. AB - Using the learned helplessness model of depression in rats, the present study undertook to investigate the possibility of an impaired response to antidepressant drugs in diabetic animals. Experimental diabetes was induced by three intraperitoneal (IP) injections of streptozotocin (37.5, 37.5, 50 mg/kg, three days apart), four weeks before behavioral testing. Diabetic and non diabetic rats were first exposed to 60 inescapable shocks. Forty-eight hours later and over three consecutive days, they were subjected to daily shuttle-box sessions for assessment of escape failures (helpless behavior). Twice daily (IP) injection of clomipramine (24 mg/kg), desipramine (24 mg/kg), imipramine (32 mg/kg) or clenbuterol (0.75 mg/kg) prevented escape deficits in the non-diabetic but not in the diabetic rats. However, this prevention was made possible in the diabetic rats by increasing the duration of the antidepressant treatment. Moreover, one week of insulin therapy restored operant escape responding to both the tricyclics and a beta-agonist. The inefficacy of clenbuterol (a central beta agonist) in reversing helpless behavior in diabetic rats, along with the observation that triiodothyronine (T3) supplementation also restored the response to imipramine in the diabetic rats, suggests that thyroid-mediated alterations of central noradrenergic function might be a critical factor in the resistance or delayed response to antidepressants in experimental diabetes. These animal findings raise the possibility of a similar resistance to conventional antidepressants in depressed diabetic patients. PMID- 2544020 TI - [Gigantic esophageal ulcer associated with a cytomegalovirus in an HIV positive patient]. AB - The authors report and HIV positive patient whose first manifestation of AIDS was the appearance of a giant esophageal ulcer due to cytomegalovirus. We comment on the frequency, clinical manifestations, diagnosis and endoscopic aspects of esophageal cytomegalovirus affectation. PMID- 2544019 TI - [Liver transplant in fibrolamellar hepatocarcinoma: apropos of a case. Review of the literature]. AB - A case is presented of fibrolamellar hepatocarcinoma in a 12 year-old male treated by liver transplantation; there is no evidence of tumoral recurrence at 28 months. Donos and receptor were ABO incompatible. The immunosuppressive regimen used was cyclosporine A and low doses of steroids. Fibrolamellar hepatocarcinoma is an infrequent histologic variety that usually affects young people and is generally not associated with hepatitis B infection or cirrhosis. It is often a single tumor, is more susceptible to surgical resection than other varieties of hepatocarcinoma, and is characterized by a relatively unagressive tumoral biology. PMID- 2543990 TI - Caffeine-induced anxiogenesis: the role of adenosine, benzodiazepine and noradrenergic receptors. AB - The purpose of this study was to determine the mechanism by which caffeine increases anxiety. Rats were tested in the social interaction test of anxiety after administration of caffeine (20 or 40 mg/kg) alone or in combination with various compounds. In order to investigate the role of adenosine receptors, caffeine was given in combination with 2-chloroadenosine (0.1 and 1 mg/kg). To investigate the role of benzodiazepine receptors, chlordiazepoxide (5 mg/kg), a benzodiazepine antagonist, flumazenil (RO 15-1788, 1 and 10 mg/kg) and a triazolobenzodiazepine U-43,465 (32 mg/kg) were used. Finally, an alpha 2 receptor agonist, clonidine (0.1 and 0.025 mg/kg) and a beta-adrenoceptor antagonist, DL-propranolol (5 mg/kg), were used to study the role of noradrenergic systems in the effects of caffeine. Caffeine (20 and 40 mg/kg) reduced the time spent in social interaction and this effect was antagonized by chlordiazepoxide, U-43,465 and DL-propranolol, but not by flumazenil, 2 chloroadenosine or clonidine. It was therefore concluded that the anxiogenic effect of caffeine was unlikely to be due to its effects at adenosine or benzodiazepine receptors. It is suggested that the reversal of caffeine's effects by chordiazepoxide may have been "functional," i.e., merely a cancellation of two opposite effects. It is discussed whether the reversal of caffeine's effects by propranolol and U-43,465 are functional, or reflect a noradrenergic site of action. PMID- 2544021 TI - [Mixed malignant tumor of the liver]. PMID- 2544022 TI - [Vegetable fiber (III). Metabolic diseases]. PMID- 2543986 TI - Visual experience regulates gene expression in the developing striate cortex. AB - We have examined the regulation of expression of the genes for the neuronal growth-associated protein GAP43, the type II calcium/calmodulin-dependent protein kinase, and glutamic acid decarboxylase in the kitten visual cortex during normal postnatal development and after a period of visual deprivation. We find that the mRNA transcripts of these genes display very different patterns of normal development but are all increased in the visual cortex of animals reared in the dark. Upon exposure to light, the transcript of the GAP43 gene drops to near normal levels within 12 hr. PMID- 2544023 TI - [Imuthiol and human immunodeficiency virus infection]. PMID- 2543997 TI - Endocrinological studies in alcoholics during withdrawal and after abstinence. AB - Several endocrine parameters were assessed in 35 alcoholic inpatients after admission to hospital, and 17 of the 35 were retested after several weeks of sobriety. No difference was found in clonidine-stimulated growth hormone (GH) secretion between male alcoholics and male healthy controls, but significant positive correlations of GH secretion and alcohol content in expired breath on admission and gamma-GT values after abstinence were observed. Nonsuppression in the dexamethasone suppression test was found in 17% of the patients on admission, which seemed to be due to alcohol withdrawal. Postdexamethasone cortisol levels were significantly positively correlated with the "apathic syndrome" (r = 0.40; p less than or equal to 0.05). About one-third of the patients showed a blunted response in the TRH-test both on admission and after abstinence. No significant influence of alcohol intake, withdrawal or familial disposition on prolactin values could be detected. The results of the TRH test and the DST point to similar endocrinological patterns in alcoholics as in depressive patients and thus support the hypothesis of a link between alcoholism and depression. PMID- 2544026 TI - Control of gene expression by artificial introns in Saccharomyces cerevisiae. AB - Artificial yeast introns that show cold-sensitive splicing have been constructed. These conditional introns can be inserted into a target gene as an "intron cassette" without disrupting the coding information, allowing expression of the gene to be cold sensitive. Insertion of these intron cassettes rendered the yeast URA3 gene cold sensitive in its expression. The advantage of this intron-mediated control system is that any gene can be converted to a controllable gene by simple insertion of an intron. PMID- 2544024 TI - Effect of human faecal donor on in vitro fermentation variables. AB - Homogenized and diluted human faeces (66.6 g/l) collected from six individuals were incubated with four standard substrates (oat bran, wheat bran, kidney beans (Phaseolus vulgaris), and guar gum) for 4, 8, 12, and 24 h. Neutral detergent fibre (NDF) digestibility coefficients, short-chain fatty acid (SCFA) production (mmol/g organic matter (OM], and gas production (ml/g OM) were significantly affected by donor, substrate, and donor X substrate interactions, within an incubation time, but substrate fermentability rankings were similar at 24 h for all six donors. Substrate differed in NDF digestibilities at all incubation times, whereas SCFA and gas productions differed only at 8, 12, and 24 h. The results indicate that inoculum source (donor) can influence in vitro fermentation variables but not the substrate fermentability ranking at 24 h. It is recommended that several donors, at least three, be used to improve the accuracy of in vitro estimates of colonic fermentation. PMID- 2544025 TI - Defective phagocytosis, decreased tumour necrosis factor-alpha production, and lymphocyte hyporesponsiveness predispose patients with systemic lupus erythematosus to infections. AB - Twenty-three patients with systemic lupus erythematosus (SLE) were studied in order to understand the mechanism of increased susceptibility to infection in SLE patients. We found that phagocytosis by polymorphonuclear leucocytes (PMN) was significantly defective in untreated (24.2 +/- 3.1%) and immunosuppressant treated SLE patients (30.0 +/- 3.6%) compared with normals (47.9 +/- 0.6%), while the generation of superoxide anion radicals was normal. The defective phagocytosis in SLE could be increased by human recombinant tumour necrosis factor alpha (TNF-alpha). However, the percentages of phagocytosis in SLE before and after TNF-alpha stimulation were 56.6% and 60.7% of the normal values. This indicates that certain populations of PMN in SLE are not only defective as regards phagocytosis but also unresponsive to TNF-alpha stimulation. In an ELISA, TNF-alpha production by phorbol myristate acetate (PMA)-stimulated mononuclear cells from SLE patients was significantly decreased (181.4 +/- 22.7 pg/ml vs. 533.0 +/- 81.9 pg/ml, p = 0.002). In addition, the percentage of phytohaemagglutinin (PHA)-stimulated mononuclear cells in S phase in the cell cycle was deficient in patients with SLE (17.2 +/- 1.8% vs. 29.7 +/- 2.9%, p less than 0.001). These results lead us to propose that defective PMN in spontaneous and TNF-alpha-induced phagocytosis, decreased production of TNF-alpha, and lymphocyte hyporesponsiveness predispose patients with SLE to infections. PMID- 2544027 TI - Isolation of single-copy human genes from a library of yeast artificial chromosome clones. AB - A recently developed cloning system based on the propagation of large DNA molecules as linear, artificial chromosomes in the yeast Saccharomyces cerevisiae provides a potential method of cloning the entire human genome in segments of several hundred kilobase pairs. Most application of this system will require the ability to recover specific sequences from libraries of yeast artificial chromosome clones and to propagate these sequences in yeast without alterations. Two single-copy genes have now been cloned from a library of yeast artificial chromosome clones that was prepared from total human DNA. Multiple, independent isolates were obtained of the genes encoding factor IX and plasminogen activator inhibitor type 2. The clones, which ranged in size from 60 to 650 kilobases, were stable on prolonged propagation in yeast and appear to contain faithful replicas of human DNA. PMID- 2543989 TI - The benzodiazepine receptor inverse agonist RO15-4513 exacerbates, but does not precipitate, ethanol withdrawal in mice. AB - RO15-4513, an imidazobenzodiazepine that has been reported to antagonize several behavioral and biochemical actions of ethanol, was given to C3H mice at various times during withdrawal from chronic (72 hours) continuous exposure to ethanol vapor. When administered immediately following chronic ethanol exposure, RO15 4513 (6 or 12 mg/kg) did not influence the withdrawal response. However, when given at subsequent times (3, 5, and 8 hours postethanol withdrawal), RO15-4513 significantly increased the severity of the withdrawal response in ethanol exposed mice. Moreover, this exacerbation was completely reversed by pretreatment with the benzodiazepine receptor antagonist RO15-1788. Thus, these data indicate that the benzodiazepine inverse agonist, RO15-4513, is capable of exacerbating, but not precipitating, ethanol withdrawal. PMID- 2543988 TI - Noradrenergic receptor interactions in feeding elicited by stimulation of the para-ventricular hypothalamus. AB - Food intake and feeding behaviour were examined after the administration of noradrenaline (NA) or clonidine to the para-ventricular nucleus of the hypothalamus, or after systemic administration of clonidine. In 20-hr deprived animals all treatments dose-dependently reduced pellet consumption; however, at a low dose (2-5 micrograms) central clonidine increased eating time and bout length. In 4-hr deprived animals all treatments increased sucrose consumption. Clonidine (peripheral and central) increased feeding time but did not alter feeding rate; NA did not alter feeding time, but did increase feeding rate; NA also increased activity and decreased resting. The effects of NA on feeding rate, activity and resting were blocked by propranolol; however, the propranolol-NA combination increased feeding time. Thus, NA and clonidine increased feeding by different mechanisms, but after propranolol pretreatment the effects of NA were similar to those of clonidine. It is concluded that clonidine enhances feeding by inhibiting satiety and that the feeding stimulant effect of NA is mediated by a complex interaction of alpha- and beta-receptors. PMID- 2544028 TI - The basic defect in cystic fibrosis. PMID- 2544029 TI - DNA looping generated by DNA bending protein IHF and the two domains of lambda integrase. AB - The multiprotein-DNA complexes that participate in bacteriophage lambda site specific recombination were used to study the combined effect of protein-induced bending and protein-mediated looping of DNA. The protein integrase (Int) is a monomer with two autonomous DNA binding domains of different sequence specificity. Stimulation of Int binding and cleavage at the low affinity core type DNA sites required interactions with the high affinity arm-type sites and depended on simultaneous binding of the sequence-specific DNA bending protein IHF (integration host factor). The bivalent DNA binding protein is positioned at high affinity sites and directed, by a DNA bending protein, to interactions with distant lower affinity sites. Assembly of this complex is independent of protein protein interactions. PMID- 2544030 TI - Degradation of proteins with acetylated amino termini by the ubiquitin system. AB - A free NH2-terminal group has been previously shown to be an obligatory signal for recognition and subsequent degradation of proteins in a partially fractionated and reconstituted ubiquitin proteolytic system. Naturally occurring proteins with acetylated NH2-termini--most cellular proteins fall in this category--were not degraded by this system. Other studies have suggested that the identity of the NH2-terminal residue is important in determining the metabolic stability of a protein in vivo (N-end rule). Whole reticulocyte lysate and antibodies directed against the ubiquitin-activating enzyme (E1) have now been used to show that such acetylated proteins are degraded in a ubiquitin-dependent mode. Although fractionation of lysate does not affect its proteolytic activity toward substrates with free NH2-termini, it completely abolishes the activity toward the blocked substrates, indicating that an important component of the system was either removed or inactivated during fractionation. An NH2-terminal "unblocking" activity that removes the blocking group, thus exposing a free NH2 terminus for recognition according to the N-end rule, does not seem to participate in this pathway. Incubation of whole lysate with labeled histone H2A results in the formation of multiple ubiquitin conjugates. In contrast, the fractionated system is devoid of any significant conjugating activity. These results suggest that a novel conjugating enzyme (possibly a ubiquitin-protein ligase) may be responsible for the degradation of these acetylated proteins by recognizing structural features of the substrate that are downstream and distinct from the NH2-terminal residue. PMID- 2544031 TI - Effects of tiaprofenic acid on interleukin 1, phospholipase A2 activity, prostaglandins, neutral protease, and collagenase activity in rheumatoid synovial fluid. AB - IL-1 and prostaglandin (PGE2, PGF2 alpha, TXB2) concentrations, PLA2 activity, neutral protease activity, and collagenase activity specific for types I and II collagen were determined in the SF of patients suffering from RA, before and after treatment with TA. Active and latent forms of protease and collagenases were regularly detected but were unrelated to IL-1, PLA2, and PGE2. TA induced a significant decrease in tested eicosanoids but IL-1, PLA2, and proteases were unchanged. PMID- 2544015 TI - The use of molecularly tagged P elements to monitor spontaneous and induced frequencies of transposon excision and transposition. PMID- 2543993 TI - Phencyclidine/SKF-10,047 binding sites: evaluation of function. AB - Results of correlation analyses comparing rank-order affinities with rank-order potencies of (+)SKF-10,047, phencyclidine (PCP), and several PCP analogs support the involvement of [3H]-1-[1-(2-thienyl)cyclohexyl]piperidine binding sites (TCP sites) in mediating both the discriminative stimulus properties of PCP and production of 180 degrees perseveration in a 4-arm radial maze. For the same group of drugs, no significant relationship was found to exist between affinities at haloperidol-sensitive (+)[3H]SKF-10,047 binding sites (H-S-SKF sites) and potencies. Also, H-S-SKF sites were found to lack pharmacological selectivity and to be localized in the microsomal fraction of cells. It is concluded that TCP sites may represent receptors which mediate effects not only of PCP, but also of (+)SKF-10,047. In addition, the possibility that H-S-SKF sites may represent a type of membrane-bound enzyme is discussed. PMID- 2543987 TI - Risk behavior-based model of the cubic growth of acquired immunodeficiency syndrome in the United States. AB - The cumulative number of cases of acquired immunodeficiency syndrome (AIDS) in the United States has grown as the cube of time rather than exponentially. We explain this by interactions involving partner choice and sexual frequency in a risk-behavior model with biased mixing. This leads to a saturation wave of infection moving from high- to low-risk groups. If this description is correct, then the decreasing growth rate of AIDS cases is not due to behavior changes; rather it is due to the intrinsic epidemiology of the disease. PMID- 2544032 TI - Molecular basis for the action of tiaprofenic acid on human osteoarthritic cartilage degradation. PMID- 2544033 TI - Interleukin 1, tumor necrosis factor, and interleukin 6 as mediators of cartilage destruction. PMID- 2544034 TI - Fine needle aspiration cytology of hepatocellular carcinoma manifested as bone metastasis. AB - Symptomatic skeletal metastasis in hepatocellular carcinoma is rare. We have presented two cases of expansile, lytic lesions involving the distal radius and sacrococcyx, respectively. Initially, diagnosis was made by fine needle aspiration cytology, complemented by positive alpha-fetoprotein immunoperoxidase staining in one and ultrastructural evaluation in both. PMID- 2544035 TI - [Peripheral neuropathy and cyclosporin. Apropos of 2 cases]. AB - We report two cases of peripheral neuropathy arose during treatment with cyclosporine. The first observation was in a 24 year-old female treated because of a recent diabetes mellitus, the second in a 52 year-old male whose treatment was given after a cardiac transplantation. Chronology, clinical presentation and morphological findings on nerve biopsy were very close in both cases ans intrinsic imputability was stated as "possible" according to the method used to assess unexpected or toxic drug reaction by the French Regional Drug Monitoring Centers. It remains to confirm with similar reports the putative part played by cyclosporine as strongly suggested in ours. PMID- 2544036 TI - [Dysgeusia during treatment with enalapril]. PMID- 2544038 TI - Protein C inhibitor: purification and proteinase reactivity. AB - Protein C inhibitor was purified from human plasma by a modification of a published procedure (Suzuki, K., Nishioka, J., and Hashimoto, S. J. Biol. Chem. 258, 163-168, 1983). Approximately 1 mg of pure protein was obtained from 1 L plasma, a yield of about 17%. The protein C inhibitor preparation did not lose activity over 4 weeks at 4 degrees C. Second order rate constants were measured for the inhibition of activated protein C, thrombin, and urokinase, and bimolecular complexes of protein C inhibitor with activated protein C and thrombin were visualized by denaturing polyacrylamide gel electrophoresis. Heparin accelerated the inhibition of the three proteinases in a manner consistent with a template mechanism. Plasma or pure protein C inhibitor (at the same concentration) showed the same effect of heparin on activated protein C inhibition, indicating that protein C inhibitor accounts for all the heparin dependent inhibition of activated protein C in vivo. PMID- 2544037 TI - Influence of phospholipase A2 on human blood platelet alpha adrenergic receptor function. AB - Exposure of platelets to phospholipase A2 selectively obliterates the aggregation response of human platelets to the action of epinephrine. This treatment does not impair the affinity of the ligand binding although it reduces the number of sites available on platelets. Reduction in the number of alpha receptors caused by phospholipase treatment in no way affected the epinephrine-induced restoration of sensitivity of refractory platelets to the action of agonists. PMID- 2544039 TI - Kinetic study of tPA release induced by heparin and heparin fragment. PMID- 2544041 TI - Effects of mercuric chloride on [3H]dopamine release from rat brain striatal synaptosomes. AB - Electrophysiological studies employing amphibian neuromuscular preparations have shown that mercuric chloride (HgCl2) in vitro increases both spontaneous and evoked neurotransmitter release. The present study examines the effect of HgCl2 on the release of [3H]dopamine from synaptosomes prepared from mammalian brain tissue. Mercuric chloride (3-10 microM) produces a concentration-dependent increase in spontaneous [3H]dopamine release from "purified" rat striatal synaptosomes, in both the presence and absence of extra-synaptosomal calcium. The effects of HgCl2 on transmitter release from amphibian neuromuscular junction preparations resemble those produced by the Na+, K+-ATPase inhibitor ouabain. Experiments were performed to determine whether the HgCl2 effects on mammalian synaptosomal dopamine release are a consequence of Na+, K+-ATPase inhibition. Na+, K+-ATPase activity in lysed synaptosomal membranes is inhibited by HgCl2 (IC50 = 160 nM). However, mercuric chloride in the presence of 1 mM ouabain still increased [3H]dopamine release. The specific inhibitor of Na+-dependent, high affinity dopamine transport, RMI81,182 inhibited ouabain-induced [3H]dopamine release whereas it had no effect on HgCl2-induced [3H]dopamine release. These data suggest that augmentation of spontaneous [3H]dopamine release by HgCl2 probably is not mediated by an inhibition of Na+, K+-ATPase and HgCl2 does not act directly on the dopamine transporter. PMID- 2544040 TI - Characterization of the oxidant generation by inflammatory cells lavaged from rat lungs following acute exposure to ozone. AB - Following exposure to 2 ppm ozone for 4 hr, two distinct effects on rat lung inflammatory cell oxidant generation were observed. TPA- and opsonized zymosan stimulated superoxide production by the inflammatory cell population was found to be maximally inhibited 24 hr following ozone exposure. In contrast, luminol amplified chemiluminescence increased 24 hr following ozone exposure, coinciding with an increase in the percentage of neutrophils and myeloperoxidase in the inflammatory cell population. Supporting the involvement of myeloperoxidase in the enhanced oxidant-generating status of these cells, the luminol-amplified chemiluminescence was found to be azide-, but not superoxide dismutase inhibitable. Additionally, this cell population was found to generate taurine chloramines, a myeloperoxidase-dependent function which was absent prior to the ozone exposure and also demonstrated enhanced activation of benzo[a]pyrene-7,8 dihydrodiol to its light-emitting dioxetane intermediate. Addition of myeloperoxidase to control alveolar macrophages resulted in enhanced luminol amplified chemiluminescence, taurine chloramine generation, and enhanced chemiluminescence from benzo[a]pyrene-7,8-dihydrodiol demonstrating that, in the presence of myeloperoxidase, alveolar macrophages are capable of supporting myeloperoxidase-dependent reactions. The possibility of such an interaction occurring in vivo is suggested by the detection of myeloperoxidase activity in the cell-free lavagates of ozone-exposed rats. These studies suggest that neutrophils recruited to ozone-exposed lungs alter the oxidant-generating capabilities in the lung which could further contribute to lung injury or to the metabolism of inhaled xenobiotics. PMID- 2544043 TI - Cellular distribution of cis-diamminedichloroplatinum(II)-DNA binding in rat dorsal root spinal ganglia: effect of the neuroprotecting peptide ORG.2766. AB - The in situ binding of the anticancer drug cis-diamminedichloroplatinum(II) (cisDDP) to DNA was studied in the rat dorsal root spinal ganglion (DRG), using an antiserum against cisDDP-modified calf thymus DNA in a quantitative immunocytochemical assay. Rats received a dose of cisDDP (1 mg/kg), two times a week, up to a cumulative dose of 15 mg/kg (group I) or 34 mg/kg (group II). Rats of group III were given a single dose of 15 mg/kg. Rats were killed 48 hr (groups I and II) or 6 hr (group III) after the last injection. In groups I and II cisDDP induced neurological damage was assessed by measuring both motor and sensory nerve conduction velocities (MNCV and SNCV). Whereas the MNCV was not influenced by the treatment with cisDDP, the SNCV decreased significantly. The level of cis DDP-DNA binding in DRG satellite cells equalled that in liver cells, but binding could not be shown in DRG neuron nuclei. The level of cisDDP-DNA binding in spinal cord and brain was very low. The neuroprotecting peptide ORG.2766, an ACTH4-9 analog, was given sc (10 micrograms/rat) four times a week concomitantly with cisDDP to some rats of groups I and II. ORG.2766 prevented the decrease of the SNCV, but did not change the extent of cisDDP-DNA binding in satellite or liver cells. It is concluded that the amelioration of cisDDP toxicity by ORG.2766 is not directly related to the cisDDP-DNA binding in satellite cells. PMID- 2544042 TI - Phenobarbital-induced cytosolic cytoprotective mechanisms that offset increases in NADPH cytochrome P450 reductase activity in menadione-mediated cytotoxicity. AB - Hepatocytes isolated from phenobarbital (PB)-pretreated and naive male Sprague Dawley rats were incubated with menadione under one of three oxygen conditions (0, 21, or 95% oxygen) for 3 hr. During this time, samples were drawn and assayed for lactate dehydrogenase release and trypan blue exclusion as indices of cytotoxicity. Neither parameter indicated any significant difference in menadione induced cytotoxicity between naive and PB-pretreated hepatocytes. Likewise, no difference was observed between hepatocytes incubated in 21% versus 95% O2. Consistent with the oxyradical hypothesis of menadione-induced cytotoxicity, hepatocytes incubated under 0% O2 (95:5; N2:CO2) did not exhibit any menadione cytotoxicity. Hepatic microsomes prepared from PB-pretreated rats exhibited a threefold increase in NADPH cytochrome P450 reductase activity over those of controls. Menadione-stimulated superoxide (O2-) production was twofold higher in PB pretreated versus naive liver microsomes. However, PB pretreatment failed to produce an increase in O2- production in intact hepatocytes or in hepatocytes disrupted by sonication. The failure of PB pretreatment to increase menadione induced cytotoxicity and superoxide production in either intact or sonicated hepatocytes suggests that a concomitant cytoprotective mechanism is induced as well. The data further indicate that the cytoprotective elements are located in a nonmicrosomal fraction of the cell. In support of this, we observed PB-induced increases in glutathione levels, glutathione reductase, and DT-diaphorase activities. These findings indicate that PB-induced enhancements of the hepatocellular cytoprotective mechanisms collectively compensate for the increased redox cycling mechanism, resulting in a mitigation of the anticipated increased hepatocellular cytotoxicity of menadione. PMID- 2544044 TI - Oral dosages of erythrosine and effects on TSH. PMID- 2544045 TI - Relationship of the hypothalamic-pituitary-adrenal axis with chemically induced immunomodulation. I. Stress-like response after exposure to T-2 toxin. AB - T-2 toxin (T-2), a trichothecene mycotoxin, produced by several members of the genus Fusarium is a cytotoxic feed contaminant and has been shown to by immunomodulatory. It is suspected that T-2 associated immunomodulation is mediated partly through the hypothalamic-pituitary-adrenal (HPA) axis. T-2, prepared in 4% ethanol/corn oil, was administered orally to male CD-1 mice. Endotoxemia was evident 24 h after a single, oral exposure to T-2. Blood levels of corticosterone, indicative of the stress-response, increased 24 h after T-2 exposure. Hypothalamic norepinephrine and serum corticosterone levels increased in a dose-related manner after 2 weeks of T-2 exposure. Endotoxin, detected in the serum of animals exposed to 2.5 mg/kg T-2 for 1 week, was not associated with bacteremia. Neither endotoxin nor bacteremia were detectable after 2 or 4 weeks of T-2 exposure. Exposure to 2.5 mg/kg T-2 also affected several organs. The forestomach was ulcerated, with lymphocytic infiltration, epithelial proliferation, and hyperkeratinization. Increased spleen weight was associated with a proliferative red pulp. No histological changes were observed in the enlarged liver. Gastritis has been associated with increased corticosteroid production; cortical depletion and reduced mass of the thymus are phenomena attributable to increased corticosteroid levels. An increased corticosteroid level has been associated with thymic involution leading potentially to decreased T-dependent antibody response, a known effect of T-2. PMID- 2544046 TI - Complexities of gene regulation by cAMP. PMID- 2544048 TI - [An analysis of the action of nonspecific and restriction nucleases on metaphase chromosomes in situ]. AB - Patterns of differential staining of Drosophila, mouse, rat, cattle and pig chromosomes were examined after the treatment with nucleases (DNAase I, DNAase II) and restriction enzymes (AluI, HpaII, MspI, BpE, EcoRI). The above effects depend on the species used, on the enzymes and substitution of thymine for bromodeoxyuridine in the chromosomal DNA. It is supposed that such a phenomenon may not only result from the irregular distribution of specific restriction sites along chromosomes but also depend on the specificity of supramolecular organization of the chromosomal DNA. PMID- 2544047 TI - [The distribution of glutamate receptors in cultures of the motor area of the rat cerebral cortex studied by an immunoelectron microscopic method]. AB - The aim of the study was to analyse the distribution and localization of glutamate receptors in the cultured cells of the neonatal rat motor cortex, using immunoelectron microscopic technique, and monoclonal antibodies preliminary labeled with colloidal gold. Monoclonal antibodies against glutamate-binding proteins of the adult rat cerebral cortex were produced by means of hybridization of immune splenocytes with plasmocytoma cells. It was found that monoclonal antibodies labeled with colloidal gold could reveal selectively the localization of glutamate receptors on the membranes of neurons. Glutamate receptors were detected on differentiating neuron membranes only, being absent from the glia cell surface. PMID- 2544049 TI - [Interleukin-1-like factor from human B-lymphocytes transformed by the Epstein Barr virus]. AB - The NC-37, EBV-transformed human B-lymphoblastoid cell line spontaneously produces interleukin (IL)-1-like factor, which is able to stimulate proliferation of mouse thymocytes and human fibroblasts. The IL-1-like factor production can be enhanced by prodigiozan and by the conditioned medium of human peripheral blood mononuclear cells. The NC-37 cells have a membrane-associated form of IL-1-like factor also. The molecular mass of the intracellular precursor of this factor determined by gel-filtration is 35-40 kDa, and that of the secretory form--18-20 kDa. The secretory form has the following isoelectric points: 4.5; 6.8; 7.2-8.4. PMID- 2544050 TI - [Effect of sodium, lithium and amiloride on the K+-dependent phosphatase activity of membrane preparations of Na,K-ATPase]. AB - Effects of sodium, lithium and amiloride on the ATPase reaction and on its potassium-dependent step were studied using membrane preparations of Na,K-ATPase. It was established that the addition of 70 mM NaCl or LiCl to the reaction medium diminished the hydrolysis of para-nitrophenyl phosphate (pNPP) by 70 and 40%, respectively. Amiloride (0.8 mM) inhibited activities of Na,K-ATPase and pNPPase by 50 and 15%, respectively. The higher concentrations of amiloride produced a more prominent inhibition of Na,K-ATPase, but not of pNPPase. There was no correlation between the effect of amiloride on the pNPP hydrolysis and potassium concentration in the medium. There was the additivity in the inhibition of pNPPase by 0.8 mM amiloride and sodium or lithium ions up to the concentrations of ions as high as 30 mM. A conclusion is made that the inhibition of Na,K-ATPase by amiloride is mediated through the modification of the sensitivity of the enzyme to sodium. PMID- 2544051 TI - Myoepithelioma--new concepts of histology and classification: a light and electron microscopic study. AB - Based on histological, immunohistochemical, and ultrastructural studies, it is now apparent that the modified myoepithelial cell component of pleomorphic adenomas has a considerable range of cytological features. We reasoned that myoepitheliomas could be tumors with a similar spectrum of neoplastic myoepithelium but lacking the ductal element displayed in pleomorphic adenomas. A review of available salivary gland tumors identified 40 examples based on this definition. Architecturally, these myoepitheliomas displayed either nonmyxoid (solid), myxoid (pleomorphic adenoma-like), reticular (canalicularlike), or mixed growth patterns, while cytologically the lesions were composed of spindle-type (32.5%), hyaline-type (7.5%), epithelial-type (45.0%), clear-type (2.5%), or mixed-type (12.5%) tumor cells. Electron microscopy was carried out on eight examples and detailed immunohistochemistry on two methanol-fixed cases. As a result of the current review of myoepitheliomas and the description of similar lesions in the literature, it is our contention that salivary gland myoepitheliomas are not as rare as has been purported. PMID- 2544052 TI - Anaplastic Wilms' tumor and other clinically aggressive childhood renal neoplasms: ultrastructural and immunocytochemical features. AB - The ultrastructural and immunocytochemical findings in a series of 32 rarely encountered childhood renal malignancies are reviewed. The discussion includes anaplastic Wilms' tumors, renal clear cell sarcomas, rhabdoid tumors, carcinomas, sarcomas, neuroendocrine tumors, and lymphomas. An attempt is made to assess the relative merits of these two techniques in the differential diagnosis of these often troublesome lesions. PMID- 2544053 TI - Peripheral neuroectodermal tumor of the chest wall in a 19-year-old female. AB - A small cell round tumor involving the chest wall of a 19-year-old female had features that by light microscopy were considered consistent with Ewing's tumor. At the ultrastructural level the cells contained small quantities of glycogen and had irregular dendritic processes with rare dense core granules and microtubules. Immunostaining for neuron-specific enolase was positive. The neoplasm is similar to reported cases of peripheral neuroectodermal tumor involving the chest wall of young patients. PMID- 2544054 TI - Benign adenomyoepithelioma of the breast. PMID- 2544056 TI - [Bronchial asthma caused by isocyanate in the internal/external environment]. AB - Di-isocyanates have resulted in sensitizing and bronchial asthma in 4-5% of exposed workers in industry. A case of bronchial asthma is described in which the primary sensitization occurred during the period 1968-1970 as a result of work in a factory producing polyurethane plastic foam. The patient sought other employment on account of severe asthmatic symptoms and was, thereafter, symptom free for 15 years. He was then engaged in a factory in the neighbourhood of the plastic foam factory and the asthmatic symptoms recurred on account of exposure via the external environment. The influences of various public authorities on handling of this case are described. PMID- 2544055 TI - Myxoid malignant fibrous histiocytoma with erythrophagocytosis. AB - A tumor that formed multiple protuberant masses on the foot and leg of a 50-year old white male had the light and electron microscopic features of a myxoid malignant fibrous histiocytoma. Ultrastructurally, the cells had some fibroblastic features. Extravasated erythrocytes were present throughout the tumor nodules and many had been phagocytosed by the tumor cells. Endothelial cell markers were negative. PMID- 2544057 TI - [Anogenital HPV infections in children and sexual child abuse]. PMID- 2544058 TI - [Low-molecular weight heparin. A new agent to prevent thrombosis]. AB - The theory for employment of low molecular weight heparin and the material concerning employment of low molecular weight heparin hitherto published is reviewed. To date, it has been demonstrated that low molecular weight heparin has the same antithrombotic efficacy as unfractionated heparin and its half life is approximately twice as long. This implies that adequate thrombosis prophylaxis can be obtained with a single daily dose. PMID- 2544060 TI - [Herpes simplex infections]. AB - In spite of modern, highly effective antiviral drugs, the treatment of herpes simplex infections is still unsatisfactory. Especially recurrent infections as well as the protection of immunocompromised patients remain little solved therapeutical problems. Neither is the vaccination of herpes simplex promising; this has been shown in a multicenter study on a new herpes simplex virus envelope antigen vaccine. Future genotechnical investigations might provide new vaccines, which might become significant for a prophylactic therapy of herpes simplex. PMID- 2544059 TI - Effects of hydrostatic pressure, H2, N2, and He, on beating frequency of rat atria. AB - Hydrostatic compression to 15 MPa caused a drop in spontaneous beating frequency (BF) of isolated rat atria kept in tris solution at 37 degrees C by 30.6 +/- 7.2%. Introduction of superfusing solutions equilibrated with hydrogen (PH2: 4.9, 9, and 14 MPa, respectively), increased the BF in proportion to the hydrogen content. A hydrogen partial pressure equal to the hydrostatic pressure was calculated to reduce the bradycardia by 52.0 +/- 19.5%. Effects of nitrogen (PN2: 5 and 14 MPa) and helium (PHe: 13 and 14 MPa) were also tested. Nitrogen was found to be 1.7-2 times and helium 0.2 times as effective as hydrogen in reducing the bradycardia. Preparations compressed at 27 degrees C exhibited a more pronounced bradycardia than those kept at 37 degrees C, but 5 MPa N2 and 9 MPa H2 reversed the bradycardia to the same extent at 27 degrees C as at 37 degrees C. Tests with 4 MPa H2 showed the effect on BF to be similar, whether the gas was added during an intermediate stop in the compression (4.6 MPa) or at 10 MPa pressure. An additional hydrostatic pressure increase from 10 to 12.5 MPa eliminated the BF increase of 4 MPa hydrogen added at 10 MPa. The findings are discussed in view of the possible use of hydrogen as a breathing gas in deep sea diving. PMID- 2544061 TI - [Parvovirus infections in dermatology]. AB - Human parvovirus B19 is the causative agent of erythema infectiosum and other atypical (rubelliform) exanthemas. On account of the relatively high prevalence of B19, these infections are rather frequent. They usually occur during childhood, but also up to the age of 30, and there are various clinical manifestations. During pregnancy, B19 infection may cause hydrops of the fetus with ensuing fetal death. Anti-B19 IgM and IgG can be detected by means of ELISA or RIA; B19-DNA is identified by hybridization of nucleic acid. PMID- 2544062 TI - [Circadian behavior of blood pressure and heart rate following orthotopic heart transplantation. Studies before and during antihypertensive therapy]. AB - The de novo hypertension, which develops in most cardiac transplant recipients within the first postoperative months, is multicausal, though toxic side-effects of cyclosporin A seem to play a key role. In order to analyze the circadian behavior of arterial blood pressure and heart rate after cardiac transplantation (HTX) and to evaluate the effect of an antihypertensive regimen on these parameters, 24-h noninvasive ambulatory blood pressure and heart rate monitoring was performed in 10 hypertensive cardiac transplant recipients on cyclosporin A (mean age 42.3 +/- 11.2 years, 14.3 +/- 8.3 months after HTX) before antihypertensive therapy and after introduction of an antihypertensive regimen with the ACE-inhibitor enalapril plus furosemide alone or combined with verapamil. The study demonstrated a complete loss of the usual nocturnal decline in blood pressure in cardiac transplant recipients (mean systolic and diastolic blood pressure 149 +/- 8 and 102 +/- 7 mm Hg during daytime and 152 +/- 8 and 104 +/- 9 mmHg at night). Antihypertensive therapy lowered the blood pressure level effectively, but did not influence the circadian pattern (mean systolic and diastolic blood pressure 121 +/- 8 and 81 +/- 4 mmHg during daytime and 121 +/- 9 and 83 +/- 3 mmHg at night, all p less than or equal to 0.001). Heart rate, in contrast, showed a significant, though in comparison to normal, a blunted decrease at night (mean heart rate 94 +/- 6 beats per min during daytime and 84 +/- 8 beats per min at night, p less than or equal to 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544063 TI - [Objective assessment of biological aging in the human: results]. AB - From a population sample of 139 healthy persons, six specially selected aging parameters were tested for their significance to vitality, by means of four mathematical models referring to the concept of vitality. As reference values, tm, is the age expressed in years when maximal vitality occurs, and the aging constant is beta, also known as an aging factor. Our results for tm extended from 16.6 to 39.6 years, depending on the respective mathematical model applied. We find a similar characteristic for the beta-values. A comparison with results in other publications on the subject, obtained by processing a considerably larger number of variables than we did, underlined the utility of our parameters. This is also shown by the quality of results we obtained by regression analysis, by the adjustment to the functions of the vitality concept and by the simplicity of the required laboratory methods of analysis. PMID- 2544064 TI - [Pain and the spine. A review. 1. On the pathophysiology of radicular pain syndromes. Current concepts explaining pain in nerve entrapment syndromes]. AB - The problems of nociception and the development of pain in radicular syndromes of the spine may be summarized as follows: 1. The anatomic complex nerve root/dorsal ganglion is a key structure in the pain physiology of disk prolapse and spinal stenosis. 2. Chronic compression of nerve tissue causes structural changes. 3. These structural changes are associated with a change in the electrical properties of the nerve root membrane. 4. The change in membrane properties in the nociceptive pathways of nerve roots is one of the preconditions for perception of pain. As the authors' remark indicate, a better understanding of the pathophysiology of pain will lead to a more differentiated therapeutic approach. PMID- 2544065 TI - [A comparative analysis of the effect of an analog of vasopressin on functionally different neurons in the grape snail]. AB - Application of desglycine-argininvasopressin (DG-AVP) differently influenced different types of cells of snail isolated central nervous system. In neurosecretory cells an increase of spontaneous impulse activity took place and, as a rule, bursts of impulses appeared, most often of synaptic origin, excluding PPa1 neurones and one of the neurosecretory cells of the left parietal ganglion. The increase of the bursts activity in these cells was based on the increase of the amplitude of membrane potential waves. Under the influence of neurosecretory cells system activation, EPSPs frequency and amplitude in secondary-sensory neurones increased, which led to a greater probability of the action potentials appearance. At prolonged action the spontaneous EPSPs in these cells began to group in bursts. Excitability and membrane resistance of these cells remained unchanged. DG-AVP had no influence on primary-sensory neurones and motoneurones. PMID- 2544066 TI - [The receptor organization of the reinforcing and analgetic opiate systems in the rat]. AB - Comparative study of topographic and receptor selectivity of emotionally positive (place preference test) and analgetic (electrical and pressure nociceptive stimulation of the tail) effects of opioids was performed in rats. Morphine and selective agonists of mu-, kappa-, delta- and sigma-opiate receptors were administered through cannulae implanted into the periaqueductal grey and ventral tegmental area (VTA). Reinforcing effects of opioids in VTA was shown to be mediated mainly by mu-, delta- and sigma-receptors, while analgetic effect was realised with the aid of mu- and delta-opioid receptors. PMID- 2544067 TI - [Hepatoblastoma: a catamnestic study and proposal of a therapy concept]. AB - Since 1977, 28 infants and children with hepatoblastomas were seen in the Department of Paediatric Surgery, Hanover Medical School. Five tumours could only be biopsied. One 11-months-old infant had a successful liver transplantation done at the Dept. of Transplantation Surgery (Professor R. Pichlmayr). Following a biopsy for unresectable tumour, chemotherapy was instituted in 5 cases: three of these could be resected in a second look operation 10 to 26 weeks later. Altogether, 22 tumours (= 81%) were resected: 14 had an extended resection (trisegmentectomy), and 8 had a lobectomy. Operative mortality was 14%. Out of 27 patients which were operated upon (resection or biopsy) longer than one year ago, 11 are alive and free from disease (= 40%). Early metastatic disease was diagnosed 3 weeks to 5 months postoperatively in 5 cases: only one of these children survived, following an intensive course of chemotherapy. In another two patients lung metastases were found 14 resp. 20 months after operation: both of these are alive and free from disease following chemotherapy and thoracotomy. One patient died of brain metastases 3 years postoperatively. Results of therapy are correlated to primary findings, type of operation, chemotherapy and histological subtype of the tumour. In comparison with published data a possible advantage of more aggressive operative strategies is discussed, and the benefit of a preoperative chemotherapy is evaluated. As a result of these considerations a treatment protocol for hepatoblastomas is proposed, designed as a national multicentre prospective study conducted by the German Society of Paediatric Oncology (GPO).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544068 TI - Cannabis plants illicitly grown in Jutland (Denmark). AB - Four hundred forty-nine fresh cannabis plants and 26 fruiting tops harvested in Jutland (Denmark) from July to September 1988 were characterized according to weight, height, marihuana yield, and cannabinoid content. The median weights were 308 g and 584 g for plants grown outdoors (n = 418) and in greenhouses (n = 31), respectively. The average marihuana yield was 8.7% for the plants grown outdoors and slightly lower for the greenhouse plants. Great variations, however, were seen both between and within the individual harvests. The mean concentration of total THC (tetrahydrocannabinol) was 0.87% for the plants grown outdoors. An increase according to the month of harvest was observed. For plants grown in greenhouses the mean value of total THC was 1.35%, while the mean concentration of fruiting tops was 2.13%. All plants contained cannabidiol (CBD), but only negligible concentrations of other cannabinoids. In approximately 80% of the plants the THC content was higher than the CBD content (drug type), while the rest either contained equal concentrations (intermediate type) or most CBD (fiber type). PMID- 2544069 TI - [The effect of gram-negative bacteria lipopolysaccharides on the development of Rauscher leukosis in BALB/c mice]. AB - The dose-dependent action of Shigella sonnei lipopolysaccharide (LPS) on the development of acute erythroleukocytosis, as well as Rauscher chronic myeloid and lymphoid leukosis, in BALB/c mice sensitive to Rauscher virus was shown. Bordetella pertussis LPS in the doses used in this investigation stimulated the development of both acute erythroleukosis and chronic myeloid and lymphoid leukosis in BALB/c mice infected with Rauscher virus. Lipid A isolated from B. pertussis LPS was found to produce a stimulating effect on the development of Rauscher leukosis in mice. After the treatment of B. pertussis LPS with polymyxin B blocking lipid A no stimulating effect of B. pertussis LPS on the development of Rauscher leukosis was observed. A suggestion is made that lipid A is the active principle contributing to the stimulation of the development of Rauscher leukosis in BALB/c mice. PMID- 2544070 TI - Infective endocarditis with inflammatory lesions in the peripheral nervous system. AB - A 64-year-old woman developed septicemia and a generalized peripheral neuropathy while being ventilated postoperatively. No cause for the neuropathy could be found during life. At autopsy she was found to have infective endocarditis and multifocal inflammatory lesions in the central and peripheral nervous systems, consistent with damage due to septic emboli. Infective endocarditis may be a cause of a generalized polyradiculoneuropathy and could be responsible for a proportion of cases of "critical illness polyneuropathy". PMID- 2544071 TI - Acute mononeuropathy with angiotropic lymphoma. AB - Angiotropic lymphoma can present as a vascular disease in the central nervous system. The patient described in this report had a sudden pain in the region of the right superficial peroneal nerve and a nerve biopsy showed tumoral cells in the lumen of most small blood vessels. This pathology, first described in the skin as malignant angioendotheliomatosis, can be compared with the occurrence of multiple emboli in the vasa nervorum. In recent cases, tumoral markers have evidenced a lymphomatous origin, generally of the B type. PMID- 2544072 TI - Neuropathy in IgM lambda paraproteinemia. Immunoreactivity to neural proteins and chondroitin sulfate. AB - In axonal neuropathies associated with IgM paraproteinemia, reports of antigen specificity of the M-protein are few. A patient with IgM paraproteinemia presented with progressive mononeuritis multiplex. IgM was found deposited in striking amounts in endoneurium and shown to bind specifically to neural proteins and chondroitin sulfates. Direct immune mechanisms, as well as the physical effects of IgM deposition, likely contributed to the development of the neuropathy. PMID- 2544074 TI - Ion-pair HPLC study of sulfonamides, barbiturates and antiphlogistic acids on a dynamically modified silica gel stationary phase. AB - The behaviour of some groups of acidic drugs (barbital derivatives, sulfonamides and antiphlogistic acids) was studied with a reversed-phase ion-pair HPLC method. A methanol-water-posphate buffered (pH = 7) mobile phase containing various concentrations of cetrimide was applied on a silica gel column. The correlations between the migration and the chemical structure within the different groups of compounds are discussed. Monofunctional benzene derivatives were utilized to investigate the influence of the various functional groups in this system on the retention. PMID- 2544073 TI - Calcineurin immunoreactivity in striatonigral degeneration. AB - The basal ganglia (including substantia nigra) of two patients with striatonigral degeneration, who had clinical histories of Parkinson's disease, were studied immunohistochemically using a purified antibody to calcineurin (CaN). Marked loss of CaN-immunoreactive neurons in the putamen and neuromelanin-pigmented neurons in the zona compacta of the substantia nigra was seen in both cases. A small number of CaN-immunoreactive neurons remained dispersed in "clusters" or "islands" in the medial portion of the putamen. In one case there was loss of CaN immunoreactive neurons in the caudate nucleus to a lesser degree than that in the putamen. In addition, both cases showed marked depletion of CaN-immunoreactive afferent nerve fibers in the external and internal segments of the globus pallidus and the zona reticulata of the substantia nigra. This report emphasizes the usefulness of the CaN-immunostaining technique for assessing the striatal efferents in human basal ganglia, and shows the neuropathological changes in the basal ganglia with striatonigral degeneration which were not possible to ascertain with previous techniques. PMID- 2544075 TI - [Ion exchange interactions on silica gel layers. II. Halogen salts of compounds containing organic tertiary and quaternary nitrogen]. AB - Primary and secondary ion exchanges--of hydrochloric acid and hydrobromic acid salts of well hydrolyzing organic bases as well as quaternary ammonium bromide which are important drug substance--taking place on silica gel using methanol as mobile phase have been investigated by thin-layer chromatographic and spectrophotometric methods. In case of tertiary ammonium salts (hydrolyzing salts) basis linked to silanate ion and halogen acid have been formed by primary ion exchange. During secondary ion exchange halogen acid has exchanged metal ions linked to silanate ions on the layer. In case of non hydrolyzing salts, the quaternary ammonium bromide salts it could not surely be proved by the applied methods whether primary ion exchange had been followed by secondary ion exchange or only primary ion exchange had occurred. PMID- 2544076 TI - [Ion exchange interactions on silica gel layers. III. Salts of organic bases with organic acids. Organic acids and sodium salts]. AB - By thin-layer chromatographic and spectroscopic (UV, IR) methods it has been proved that bass and acid are formed from salts of organic bases with organic acids during primary interaction because of the ion exchange behaviour of the silica gel layer. The organic acids formed have interacted with metals on the layer during secondary ion exchange and they migrate further as salts again. It has also been established that the extent of secondary ion exchange between metal silanate groups and acids depends on pKS values of acids. On the basis of investigations it can be ascertained with great probability that organic acids of pKS greater than 7 do not interact with metal silanate groups of silica gel layer. Sodium salts of organic acids of pKS less than 3 values similarly do not interact on the other hand sodium salts of organic acids of pKS greater than 3 values are able to interact with silanol groups. It has been established by flame atomic emission method that salicylic acid interacts with sodium ion and 86% of it migrates further as sodium salicylate. PMID- 2544077 TI - The role of tail skin temperature in the facilitation of the tail-flick reflex after spinal transection or interference with descending serotonergic neurotransmission. AB - We examined in mice whether tail skin temperatures and tail-flick reflexes were changed after spinal transection or intrathecal (i.th.) injection of the serotonin (5-HT) neurotoxin 5,6-dihydroxytryptamine (5,6-DHT) or the 5-HT receptor antagonist metergoline. Transection of the spinal cord reduced tail flick latency (the time needed to evoke the tail-flick reflex by radiant heat) and increased tail skin temperature 15-21 days after surgery. Analysis of covariance showed that the effect of tail skin temperature on tail-flick latency was far more pronounced than the effect of spinal transection. Intrathecal injection of 5,6-DHT (5 or 10 micrograms mouse-1), which extensively reduced the spinal levels of 5-HT, reduced tail-flick latency and increased tail skin temperature 1-5 days after treatment. Similarly, tail-flick latency was shortened and tail skin temperature elevated 15 min after i.th. injection of metergoline (0.5 micrograms mouse-1). Analysis of covariance showed no significant effect of i.th. 5,6-DHT or i.th. metergoline on tail-flick latency. Tail skin temperature, on the other hand, had a highly significant effect on tail-flick latency. The results show that the facilitation of the tail-flick reflex in spinally transected mice and mice injected i.th. with 5,6-DHT or metergoline is mainly caused by increased tail skin temperature. The data do not indicate that descending 5-HT pathways tonically inhibit the tail-flick reflex. PMID- 2544080 TI - Potassium channel blocking agents induce occasional spontaneous action potentials and multiquantal ATP release in sympathetic nerve terminals. PMID- 2544079 TI - Action potentials of cultured human oat cells: whole-cell measurements with the patch-clamp technique. AB - Oat cells (of the small cell carcinoma of the lung) have been reported to generate calcium action potentials. The calcium channels have further been suggested to play a crucial role in the relation between oat cell carcinoma and the often associated myasthenic syndrome. We have examined cultured human oat cells (U-1690) under voltage-clamp conditions, using the patch-clamp technique. We found, contrary to previous reports, that the action potential was caused by sodium and potassium currents. No calcium current was detected under these conditions, which indicates that calcium channels, if present, are very rare. The findings restrict, but do not rule out, the hypothesis that calcium plays a key role in the carcinoma/myasthenic syndrome relation. PMID- 2544084 TI - Drosophila retrotransposons: interactions with genome. PMID- 2544081 TI - Production and in vivo biologic actions of recombinant mouse interferon alpha 2. AB - We have expressed a recombinant mouse interferon alpha (r.Mu-IFN alpha 2) in Escherichia coli under the control of a tryptophan promoter using a synthetic adaptor formed by annealing two partially complementary oligonucleotides which introduced an ATG start codon and re-established the complete coding sequence of the mature IFN alpha 2 protein in the expression vector. Levels of up to 10(7) reference units of Mu-IFN alpha 2 per liter of culture were obtained using this construction. This recombinant mouse interferon alpha 2 exhibited antiviral activity in mice infected with EMC virus and antitumor activity in mice inoculated with Friend leukemia cells in a manner similar to that of natural mouse interferon alpha/beta. PMID- 2544082 TI - A novel concept in the purification of human leukocyte interferon. AB - A method for the purification of natural human interferon alpha (HuIFN Alpha) is described. It involves adsorption of interferon on silicic acid and its elution with hydrophobic electrolyte solution. Thereafter, elimination of possible viruses is achieved using nonionic detergent and ultrafiltration. Interferon recovered in the ultrafiltrate is further purified on Sephacryl S-200. Fractions corresponding to molecular weights ranging from approximately 10,000 to approximately 40,000 daltons are collected and directly applied on immobilized zinc affinity gel. IFN Alpha which is eluted with the non adsorbed fraction is subjected to buffer exchange, concentration, and sterilization. The resulting solution shows high specific activity (1 x 10(7) IU) with an apparently natural composition of interferon isospecies. The uniform buffer employed through the entire purification process makes it simple, fast and reproducible. PMID- 2544083 TI - Molecular pathogenesis of Sindbis virus encephalitis in experimental animals. AB - In general, the analysis of a number of strains of Sindbis virus has revealed amino acid differences of potential importance for virulence at relatively few positions in the E2-glycoprotein. Only 10 amino acid changes are potentially implicated, and 9 of these 10 lie in the N-terminal half of the protein (Fig. 1.). Currently, there is strong evidence to implicate 3 of these positions (E2 55, -114, and -172) in virulence (Table V). As more recombinant viruses are prepared and analyzed, the evidence for or against the relevance of other changes should become apparent. As is generally true in alphaviruses, the E1 gene is more invariant than E2 and analysis of several strains has revealed amino acid changes at only four positions (Fig. 2). Two, 72, and 75, are just N-terminal to the hydrophobic segment postulated to be the site of fusion activity, suggesting the possibility that virus entry into the host cell could be affected by amino acid differences at these locations. The other two changes (at 237 and 313) are distant from the fusion site on the linear molecule, but changes at 313 do affect the pH fusion suggesting participation of this site in providing stability to the glycoprotein trimers. The mechanism of altered virulence associated with any amino acid change in the E1- or E2-glycoproteins has yet to be determined. The change at E2-114 associated with reduced virulence in mice shows reduced latency and increased virulence in BHK-21 cells in vitro. This suggests that some changes result in enhanced replication that is host cell-specific. There are several points in the replication cycle of Sindbis virus where the glycoproteins and their ability to undergo conformational changes play an important role in efficiency of replication. These include attachment, fusion, transport through the Golgi, assembly, and budding from the cell surface. Some steps in replication involve host cell proteins (Baric et al., 1983), so that there may be unique, unexplored interactions with neurons or ependymal cells leading to increased neurovirulence for mice that are not represented in the typical BHK, Vero, or chick embryo fibroblast cell culture system. The task now will be to determine why specific amino changes in the proteins of Sindbis virus cause such dramatic changes in the biological properties of the virus. PMID- 2544085 TI - Pharmacological properties of the hydroxylated histamine, (+/-)-4(5)-(2-amino-1 hydroxyethyl)-imidazole. AB - The pharmacological effects of the beta-hydroxylated histamine, 4(5)-(2-amino-1 hydroxyethyl)-imidazole, on smooth muscle contraction of the ileum (H1-receptor activity) and gastric acid secretion (H2-receptor activity) of the guinea-pig were investigated and compared with those of histamine. Although beta-hydroxy histamine contracted the ileum with the same maximal response as histamine, the concentration response curve was shifted to the right by approximately three orders of magnitude. At submaximal concentrations, co-administration of beta hydroxy histamine with histamine revealed only additive effects. This H1-activity was competitively inhibited by diphenhydramine. Similarly, the hydroxylated analogue also increased intracellular cyclic AMP level and [14C] aminopyrine accumulation as a marker of acid secretion in the parietal cells. However, the EC50 was approximately ten fold that of histamine. This H2-receptor activity was inhibited completely by cimetidine. These results suggest that beta-hydroxy histamine possesses nearly full intrinsic activities at both H1 and H2-receptors and that the introduction of a hydroxyl group at the beta-carbon reduces and dissociates these activities. PMID- 2544078 TI - Sympathetic transmission in small mesenteric arteries from the rat: highly calcium-dependent at low stimulation rates. AB - In the present study we examine the calcium requirements of the neurogenic response in vitro of small arteries (150-200 microns diameter) from the mesentery of Wistar rats. Intramural nerves were activated with electrical field stimulation. Responses to single impulses and to low-frequency repeated stimulation were reduced or abolished by reducing the Ca2+ concentration in the bathing solution from 2.5 to 1.0 mM. Responses to higher frequencies (16 Hz) were only slightly affected. Since calcium reduction had markedly less effect on responses to direct activation of the smooth muscle and on responses to any dose of exogenous noradrenaline, the calcium reduction had mainly pre-junctional effects. The data show that part of the neurogenic response is highly calcium sensitive, perhaps more so than would be expected of a purely noradrenergic transmission. PMID- 2544086 TI - The interaction of the cardioprotective agent ICRF-187 [+)-1,2-bis(3,5 dioxopiperazinyl-1-yL)propane); its hydrolysis product (ICRF-198); and other chelating agents with the Fe(III) and Cu(II) complexes of adriamycin. AB - Membrane-permeable ICRF-187 [+]-1,2-bis(3,5-dioxopiperazinyl-1-yl)propane) has shown promise as a cardioprotective agent against adriamycin-induced cardiotoxicity. ICRF-187 may act through its rings-opened hydrolysis product (ICRF-198), which has an EDTA-type structure and, likewise, strongly binds metal ions. The reactions of these compounds with Fe3+-adriamycin and Cu2+-adriamycin complexes were examined. ICRF-198 quickly and completely removed both Fe3+ and Cu2+ from their complexes with adriamycin. ICRF-187 also reacted directly, but more slowly, with Fe3+-adriamycin to remove Fe3+ from the complex. This reaction was first order in ICRF-187 and Fe3+-adriamycin and yielded a second order rate constant of 123 M-1 min-1. Metal ion-complex promoted hydrolysis may thus contribute to the in vivo hydrolysis of ICRF-187 to its metal ion-chelating active rings-opened form. Both ICRF-187 and ICRF-198 were very effective in preventing the Fe3+-adriamycin induced inactivation of the cytochrome c oxidase activity of submitochondrial particles. A number of other chelating agents (desferal; penicillamine; DTPA; EDTA; TPEN; bathophenanthroline sulfonic acid; 2,2'-bipyridine; 1.10-phenanthroline, glutathione and 2-mercaptoethanol) were also examined for their ability to remove Fe3+ and Cu2+ from their complexes with adriamycin. PMID- 2544087 TI - [Clinical effects of intravesical instillation of polymyxin B in patients with acute cystitis]. AB - The effectiveness of intravesical instillation of Polymyxin B was studied in 17 patients suffering from acute cystitis. The 17 patients, all female, had an average age of 42 years. The average duration between the onset of symptoms and the initial visit to the hospital was 4.6 days. The causative organisms were 15 strains of E. coli and 1 each of P. rettegeri and K. pneumonia, respectively. Improvement of symptoms was observed in 76% of the cases but pyuria was cleared in only 47% of the cases. Clinical results were excellent in 8 cases (47%), good in 6 cases (35%) and poor in 3 cases (18%). The overall effectiveness rate was 82%. No appreciable amount o Polymyxin B, was detected in the serum of 4 patients examined. PMID- 2544090 TI - Prenatal diagnosis of I-cell disease in the first and second trimesters. AB - First trimester prenatal diagnosis of I-cell disease (1 case) was based on demonstration of profound deficiency of N-acetylglucosamine 1-phosphotransferase in chorionic villi and in cultured trophoblasts derived from the chorionic villus specimen. Deficiency of this enzyme in cultured amniotic fluid cells obtained via amniocentesis was the basis for prenatal diagnosis of I-cell disease in the second trimester (2 cases). In both procedures, the diagnosis was corroborated by the finding of intracellular deficiency and extracellular elevation of multiple lysosomal enzymes in the fetal cell cultures (trophoblasts and amniotic fluid cells), as well as a significant increase in several lysosomal enzyme activities in the maternal serum. PMID- 2544088 TI - [Pneumocystis carinii pneumonia in renal transplant recipients treated with cyclosporin]. AB - Four cases of pneumocystis carinii pneumonia occurred among 38 renal transplant recipients. Diagnosis was confirmed by cyst concentration technique in 2 cases. The other 2 cases were clinically, from the rapid improvement of fever, pulmonary infiltrates and hypoxia following a therapeutic trial of high dose sulfamethoxazole-trimethoprim. All patients responded to treatment with high dose sulfamethoxazole-trimethoprim. Three patients survived pneumocystis carinii pneumonia, but i died due to aspergillosis. One patients showed serological evidence of concomitant cytomegalovirus infection. PMID- 2544089 TI - Renal and systemic effects of short-term high protein feeding in normal rats. AB - Various studies have shown that a high protein (HP) diet, compared to a low protein (LP) diet, leads to hypercalciuria and alterations in renal and systemic hemodynamics. The authors compared the effects of HP diet to those of normal protein diet (NP) to determine the possible mechanisms by which changes in systemic hemodynamics and hypercalciuria occurred. The studies were conducted in awake rats; the effects of dietary sodium content on the changes induced by HP also were evaluated. The relationship of prostaglandins (PG), renin (PRA), and aldosterone (PA) to changes in blood pressure (BP) was assessed. Two weeks after HP and normal sodium feeding (40%), glomerular filtration rate (GFR) and urine flow (V) were not different from the same values in a group on an NP diet (23%). When HP was fed with low sodium, there was a rise in V as a consequence of greater fluid intake. Although plasma calcium remained constant, the hypercalciuria correlated with high protein and sodium content. Alterations in 1,25(OH)2 vitamin D3 or PTH (cyclic AMP excretion) function did not explain the hypercalciuria induced by HP. This suggests that HP leads to inhibition of tubular calcium reabsorption by mechanism(s) yet to be elucidated. Although HP did not alter GFR, it led to an increase in BP, a fall in renal vascular resistance, and an increase in RPF, regardless of sodium intake. PRA and urine PGE2 excretion were significantly higher in the rats on HP diet, whereas PA remained unchanged.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544093 TI - Isoproterenol, DBcAMP, and forskolin inhibit cardiac sodium current. AB - We studied the effects of isoproterenol (ISP), dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP), and forskolin on the sodium current (INa) of guinea pig ventricular myocytes using the tight-seal, whole cell voltage-clamp method. The extracellular [Na+] [( Na+]o) was decreased to 60 mM by replacing NaCl with sucrose (temperature, 32-33 degrees C). Ionic currents other than Na+ were suppressed using appropriate channel blockers. Depolarizing clamp pulse (duration, 30 ms) was applied at a rate of 0.2 Hz from a holding potential of -80 mV. ISP (1 microM) decreased the peak INa by 34% from 6.1 +/- 1.9 (SD) nA (control) to 4.0 +/- 1.5 nA (n = 7). The inhibition was more prominent at less negative potentials and disappeared in the presence of a beta-blocker (10 microM atenolol). The effects of DBcAMP (1-5 mM) and forskolin (3 microM) mimicked those of ISP and depressed the peak INa reversibly. DBcAMP (5 mM) shifted the inactivation curve of INa [h infinity-membrane potential (Em) relationship] to a hyperpolarizing direction, by 3.4 +/- 0.8 mV (n = 5). These findings suggest that ISP inhibits the cardiac INa+, probably by altering the gating mechanism of the Na+ channel, and that the effect is secondary to the increased levels of intracellular cAMP, with possible acceleration of cAMP-dependent phosphorylation of the channel. PMID- 2544092 TI - Cushing's disease in two sisters. AB - Hereditary Cushing's syndrome is an uncommon clinical entity, and most reported cases have been described in families with nodular adrenocortical dysplasia. Isolated cases of Cushing's disease (pituitary-dependent bilateral adrenal hyperplasia) have been reported in association with the multiple endocrine neoplasia syndrome, Type I (MEN I), but there are no published reports of pedigrees with more than one affected family member. Within a period of 8 months, two sisters presented with clinical findings suggestive of hypercortisolism, and Cushing's disease was confirmed by appropriate diagnostic studies. There was no evidence of any other endocrine excess syndrome in either patient. Transsphenoidal pituitary surgery confirmed the presence of an ACTH immunostaining pituitary adenoma in each woman. The authors think this is the first report in the English literature of Cushing's disease in first-degree relatives. PMID- 2544094 TI - Hypoxia stimulates endothelial cell angiotensin-converting enzyme antigen synthesis. AB - Previous studies from our laboratory indicate that exposure of the rat to chronic normobaric hypoxia reduces stores of active angiotensin-converting enzyme (ACE) in the lung. This study assesses directly the effects of hypoxia on ACE synthesis in cultured porcine pulmonary artery endothelial cells. Confluent cultures were exposed to hypoxia [2.5% O2 at 1 atmosphere (atm)] in a triple gas incubator; controls were cultured in normoxic conditions. After 24-, 48-, and 72-h exposure to hypoxic or normoxic conditions, followed by incubation with [35S]methionine for an additional 24 h under the same conditions, newly synthesized radiolabeled ACE was quantitated. Radiolabeled ACE was isolated by an immunobead procedure using either anti-ACE (porcine lung) immunoglobin G (IgG) or nonimmune IgG. A single radiolabeled peak (150 kDa) with the same electrophoretic mobility as purified porcine lung ACE was observed. There was a significant time-dependent increase in endothelial cell ACE antigen synthesis without a concomitant change in either cell number or total trichloroacetic (TCA)-precipitable protein in hypoxic cells compared with normoxic controls. In contrast, ACE activity, assessed by conversion of 125I-labeled angiotensin I to 125I-labeled angiotensin II was unchanged in cultures exposed to hypoxia (2.5% O2). This suggests that an inactive form of ACE is synthesized by cultured pulmonary artery endothelial cells under hypoxic conditions. PMID- 2544095 TI - Correction of proton and Ca association constants of EGTA for temperature and ionic strength. AB - The association constants of ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N' tetraacetic acid (EGTA) for protons and Ca can be used to calculate the apparent association constant of EGTA for Ca (K'CaEGTA). This value is often used in calculating the free [Ca2+] of complex solutions such as those used to determine the Ca sensitivity of skinned muscle fibers. As association constants are usually measured at 0.1 M ionic strength and between 20 and 25 degrees C, these constants must first be adjusted for conditions different from those at which they were measured, before calculation of K'CaEGTA. The proton and Ca association constants (and their delta H values) from A. E. Martell and R. M. Smith (Critical Stability Constants, New York: Plenum, vol. 1, 1974) adjusted for ionic strength and temperature using a semiempirical Debye-Huckel formalism and Van't Hoff isochore, respectively, closely describe the influence of both ionic strength and temperature on K'CaEGTA. Errors in the adjustment or calculation of association constants can dramatically affect the calculated value of K'CaEGTA and hence the calculated free [Ca2+] of complex solutions, such as those used to mimic the intracellular environment. PMID- 2544091 TI - Studies of insulin resistance in the streptozotocin diabetic and BB rat: activation of low Km cAMP phosphodiesterase by insulin. AB - The streptozotocin diabetic rat (STZ-DM) has been the best animal model for the study of insulin-deficient diabetes. A spontaneous diabetic BB Wistar Rat (SDR) has now been evaluated as a model for insulin-dependent diabetes that more closely reflects this disease in humans. The authors assessed the ability of insulin to stimulate the Vmax of a low Km cAMP phosphodiesterase (PDE) in adipose tissue of control, streptozotocin diabetic (STZ-DM) rats, and spontaneous diabetic BB rats (SDR). In addition, the authors examined the effect of streptozotocin on the nondiabetic littermates of the SDR animal, the NDR rat. Insulin stimulated Vmax of low Km cAMP PDE in control rat adipose tissue by 20% at 5 minutes. Insulin also stimulated Vmax of both SDR and NDR by 50% at 5 minutes. In contrast to control and both subgroups of the BB rat (SDR and NDR), insulin stimulated adipose tissue from STZ-DM less than 10% at 5 minutes. NDR animals rendered diabetic with streptozotocin were more responsive to insulin. The data demonstrate some similarities and differences between streptozotocin induced diabetes and spontaneous diabetes in the BB rat. Reduced responsiveness to insulin appears to be more a part characteristic of streptozotocin diabetes than diabetes in the BB rat. The absence of significant insulin resistance in the spontaneous diabetic BB rat also is more consistent with the pathophysiological mechanisms usually seen both in other insulin-dependent diabetic rat models and insulin-dependent diabetes in man. However, both animal models of diabetes, ie, STZ-DM and BB, like man, respond to insulin therapy. PMID- 2544099 TI - Adrenergic control of renin during dietary sodium deprivation in conscious dogs. AB - These experiments evaluated the contribution of alpha- and beta-adrenergic stimulation to plasma renin activity (PRA) during early and long-term dietary sodium restriction, compared with normal sodium intake. Uninephrectomized conscious dogs with catheters in the aorta, vena cava, and remaining renal artery were studied during normal sodium diet (approximately 70 meq/day), after 2-3 days of low-sodium diet (5-7 meq/day), and after greater than or equal to 2 wk of low sodium diet. Direct renal arterial (ira) infusion of phenoxybenzamine plus propranolol decreased PRA by similar proportions (39-48%) during all three states of dietary sodium intake. The PRA achieved after adrenergic blockade remained higher (P less than 0.05) during early and long-term sodium restriction than during normal sodium intake. The effect on PRA of ira infusion of propranolol alone was not different from that of phenoxybenzamine plus propranolol during normal or low-sodium diet, and the magnitude of decrease in PRA during low-sodium diet was the same whether propranolol (1 microgram.kg-1.min-1) was infused ira or intravenously. In summary, beta-adrenergic stimulation accounts for similar proportions of PRA during early and long-term dietary sodium restriction and during normal sodium intake. Renal alpha-adrenoceptors appear to play little or no role in control of PRA under these conditions. PMID- 2544096 TI - Effect of higher aortic pressure on ribosome formation and cAMP content in rat heart. AB - Elevation of aortic perfusion pressure from 60 to 120 mmHg in beating and arrested rat hearts raised cAMP content and increased rates of ribosome formation but had no effect on total protein synthesis during 1 h of perfusion. The activity of adenosine 3',5'-cyclic monophosphate (cAMP)-dependent protein kinase increased as perfusion pressure was elevated in arrested hearts. A regulatory link between increased cAMP content and accelerated ribosome formation was hypothesized to exist. When hearts were arrested with tetrodotoxin and exposed to 0.2 mM methacholine, a muscarinic-cholinergic agonist that blocked pressure induced increases in cAMP content, elevation of aortic pressure to 120 mmHg failed to increase the rate of ribosome formation. When aortic pressure was maintained at 60 mmHg, exposure of beating hearts to glucagon increased cAMP content and mimicked the effect of elevated aortic pressure to accelerate rates of ribosome formation. These studies support the hypothesis that increased aortic pressure preferentially accelerates rates of ribosome formation by a cAMP dependent mechanism. PMID- 2544097 TI - Neuraminidase selectively enhances transient Ca2+ current in cardiac myocytes. AB - Sialic acid, an anionic sugar moiety found peripherally on membrane glycoconjugates, is specifically hydrolyzed from the cell surface by neuraminidase. Because neuraminidase has previously been demonstrated to augment myocardial cell calcium content, the effects of neuraminidase on Ca channel function were studied on voltage-clamped guinea pig ventricular myocytes. In 25 50% of cells, neuraminidase treatment (0.12 U/ml for 20 min) enhanced current through the transient (T) Ca channel by 304 +/- 35% without significantly altering the magnitude of the long-lasting (L) Ca channel current. Exposure to neuraminidase did not affect the voltage dependence of activation or inactivation, nor did it affect the selective inhibition of the T-channel current by amiloride or the L-channel current by nifedipine. After neuraminidase treatment, the T-channel current inactivated more rapidly (time constant decreasing from 8.9 +/- 0.9 to 7.7 +/- 0.6 ms), whereas there was no change in the rate of inactivation of the L-channel current. Neuraminidase treatment removed approximately 20% of the total cellular sialic acid. These results indicate that neuraminidase treatment selectively modulates the function of the T Ca channel in ventricular myocytes, possibly through removal of sarcolemmal sialic acid, suggesting that glycosylation of membrane macromolecules may influence membrane function. PMID- 2544098 TI - Ouabain treatment of cardiac cells induces enhanced Na+-Ca2+ exchange activity. AB - Inhibition of the cardiac Na+-K+-ATPase with cardiac glycosides causes a rise in internal Na+ and a subsequent increase in cellular Ca2+ due to the sarcolemmal Na+-Ca2+ exchange mechanism. We investigated the adaptation of cultured cardiac cells to prolonged elevation of internal Ca2+ after exposure to ouabain. Cultured neonatal rat heart cells were treated with 100 microM ouabain for 4-48 h. This ouabain concentration inhibited Na+-K+-ATPase activity by approximately 45% and caused modest cellular Ca2+ loading. We found that cells adapted to ouabain treatment by increasing the amount of sarcolemmal Na+-Ca2+ exchange activity by 50-90% over a 24-h period. Kinetic and immunological data indicate that the increase was due to increased numbers of functional exchangers. Neither total cellular nor total sarcolemmal protein content was affected by the ouabain treatment. These results may be relevant toward understanding the effects of therapeutic use of cardiac glycosides. PMID- 2544101 TI - Biochemical studies on opioid and alpha 2-adrenergic receptors in canine submucosal neurons. AB - Biochemical information about receptors for adrenergic and opioid neurotransmission in submucosal plexus (SMP) is unavailable. We have purified a fraction P2 enriched in synaptosomes and neuronal membranes (high [3H]saxitoxin binding and high vasoactive intestinal polypeptide immunoreactivity, low activity of 5'-nucleotidase) from the canine small intestine SMP. The synaptosomal fraction (fraction P2) also contained a high density of opioid diprenorphine binding sites of high affinity. [3H]rauwolscine binding was enriched both in fraction P2 and in a microsomal fraction. Competition experiments using several adrenergic and opioid receptor ligands revealed that opioid receptors were approximately 64% mu-, 24% delta-, and 12% kappa-subtypes and that adrenoceptors on fraction P2 were alpha 2-subtype but that there was a heterogeneous population of alpha 2-adrenoceptors. These studies show that a fraction enriched in synaptosomes and neural membranes from the canine intestine SMP contains opioid as well as alpha 2-adrenoceptors, that all three subtypes of opioid receptors seem to be present with mu-receptors predominant, and that subtypes of alpha 2 adrenoceptors appear to be present. PMID- 2544100 TI - Microflora-derived polyamines modulate obstruction-induced colonic mucosal hypertrophy. AB - Experiments were designed to determine the role of microflora-derived intraluminal polyamines in the colonic mucosal response to obstruction. Sprague Dawley rats were treated per os with 0.9% NaCl or a combination of nonabsorbable antibiotics prior to the placement of either a sham or complete colonic obstruction. Sixty-six hours after surgery, wet tissue weight, DNA, RNA, and protein content were all increased in the mucosa proximal to the obstruction in NaCl-treated animals; however, DNA content was the only parameter increased after antibiotics. This induction was a purely local effect as neither hyperplasia nor hypertrophy was observed in the ileum or colon distal to the obstruction. In the NaCl-treated animals, mucosal ornithine decarboxylase activity was not induced until 48 h postsurgery, yet mucosal spermidine concentrations were significantly higher as early as 24 h. Intraluminal bacterial lysine, ornithine, and arginine decarboxylase activities were induced by obstruction but were reduced by antibiotic treatment. [14C]putrescine uptake by intestinal epithelial cells (IEC 6) in culture was blocked by the antibiotics employed in this study, but [14C] lysine transport was relatively unaffected. These data demonstrate that intraluminal polyamines modulate the trophic response of the colonic mucosa after colonic obstruction. PMID- 2544104 TI - Primary sequence of Xenopus laevis Na+-K+-ATPase and its localization in A6 kidney cells. AB - Polyclonal antibodies raised against the alpha- and beta-subunits of amphibian kidney Na+-K+-ATPase were used to screen an expression library from Xenopus laevis kidney epithelial cells (A6 cell line). cDNAs coding for each Na+-K+ ATPase subunit were identified and used to isolate near full-length cDNAs. The complete nucleotide sequence and the deduced amino acid sequence were determined. The alpha-subunit is an alpha (alpha I)-isoform. The alpha- and beta-subunits are more closely related to the mammalian and avian than the fish sequences. Antibodies raised against the fusion proteins produced by the two clones served to immunoprecipitate proteins from biosynthetically labeled or selectively surface-radioiodinated A6 cells grown on a porous substrate. The alpha- and the beta-subunits of Na+-K+-ATPase were found associated early in the course of biosynthesis and were restricted to the basolateral plasma membrane. PMID- 2544105 TI - Effects of luminal Na+ on single Na+ channels in A6 cells, a regulatory role for protein kinase C. AB - Na+ "self-inhibition" in tight epithelia describes the reduction in apical Na+ permeability observed with increasing luminal Na+ concentration. Patch clamp was used to examine regulation of self-inhibition at the level of single Na+ channels. After cell-attached patches (pipette solution, 129 mM NaCl) were obtained on amphibian distal nephron cells (A6), the 129 mM NaCl (high Na+) apical bath outside of the patch was replaced with 3 mM NaCl (low Na+). Within minutes there was an increase in open channel probability (Po) and the appearance of one to five "new" channels in patch membranes. A similar increase occurred when apical Na+ entry was blocked by luminal amiloride (10 microM). A23187 (1 microM), a calcium ionophore, added after low Na+ exchange, abolished the rise in channel activity. Increased Po and new channels, induced by either luminal Na+ or amiloride, were also reversed by either 4B-phorbol 12-myristate 13-acetate (PMA; 0.1 microM) or 1-oleyl-2-acetyl glycerol (OAG; 10 microM) over 15-30 min. 4 alpha Phorbol (0.1 microM), an inactive phorbol, did not reduce channel activity. D Sphingosine (100 microM), a protein kinase C (PKC) inhibitor, increased Po and new channels. CONCLUSIONS: 1) modulation of apical Na+ permeability by luminal Na+ does not require direct interaction of Na+ with the channel protein but, rather, appears to involve an intracellular regulatory pathway, 2) relieving self inhibition alters both the number and kinetics of single Na+ channels, 3) the effect of low Na+ must be modulated via decreased apical Na+ entry and intracellular Na+, since amiloride yielded similar results, 4) changes in intracellular Na+ probably affect Na+ channel activity via cytosolic Ca2+, 5) the effects of decreasing luminal Na+ are reversed by PKC activators and mimicked by PKC inhibitors suggesting a possible role for PKC in Na+ self-inhibition. PMID- 2544102 TI - Distribution of opioid receptors in canine small intestine: implications for function. AB - Distribution of the binding sites for [3H]diprenorphine, a non-selective opiate ligand, was studied in membrane fractions from longitudinal muscle/myenteric plexus and circular muscle containing deep muscular plexus. [3H]saxitoxin was used as a marker for neuronal plasma membranes and 5'-nucleotidase as a marker for smooth muscle plasma membranes. Saxitoxin binding correlated strongly with diprenorphine binding, but 5'-nucleotidase correlated poorly with diprenorphine or saxitoxin binding in these fractions. Opiate binding sites in membranes of myenteric and deep muscular plexus were of high affinity (Kd = 0.12 and 0.18 nM, respectively) with maximum binding capacity of 400 and 500 fmol/mg protein, respectively. Competition experiments using subtype-selective opiate ligands indicated that all three subtypes of opiate receptors were present in the same ratio of 40-45% mu-subtypes, 40-45% delta-subtypes, and 10-15% kappa-subtypes on both plexuses. Opiate receptors of canine small intestine, therefore, are located primarily or exclusively on nerves with similar distributions in nerve membranes containing only axonal varicosities (deep muscular plexus) as in those containing neurons, dendrites, and varicosities (myenteric plexus). PMID- 2544106 TI - Response of rat inner medullary collecting duct to epidermal growth factor. AB - Urine is an abundant source of epidermal growth factor (EGF) and prepro-EGF has been localized to the thick ascending limb and distal convoluted tubule of the kidney. However, the functional role of EGF in the kidney is poorly understood. Determination of EGF receptors and functional responses to EGF in intrarenal structures distal to the site of renal EGF production may prove critical to our understanding of the role of this peptide. These studies were designed to investigate the response to EGF of rat inner medullary collecting duct cells in culture and in freshly isolated suspensions. Primary cultures of inner medullary collecting duct cells demonstrated equilibrium binding of 125I-labeled EGF at 4 and 23 degrees C. At 23 degrees C, there was 89 +/- 1% specific binding (n = 30). Scatchard analysis of 125I-EGF binding suggested the presence of both high affinity binding with a dissociation constant (Kd) of 5 X 10(-10) M and maximal binding sites (Ro) of 2.7 X 10(3) binding sites/cell and low-affinity binding, with Kd of 8.3 X 10(-9) M and Ro of 1.8 X 10(4) binding sites/cell. Bound EGF, 68 +/- 3%, was internalized by 45 min. EGF binding was not inhibited by antidiuretic hormone, atrial natriuretic peptide or bradykinin at 23 degrees C, but there was concentration-dependent inhibition of binding by transforming growth factor alpha. Incubation with phorbol myristate acetate decreased 125I-EGF binding in a concentration-dependent manner. 125I-EGF binding was also demonstrated in freshly isolated suspensions of rat inner medullary collecting duct cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544107 TI - Proton compartmentation in rat renal cortical tubules. AB - To study the control of renal ammoniagenesis, a technique was developed to estimate simultaneously intracellular (pHi) and intramitochondrial (pHm) pH in suspensions of rat renal cortical tubules. pHi was estimated with the fluorescent probe 2',7'biscarboxyethyl-5(6)-carboxy-fluorescein (BCECF). The intracellular distribution of the weak acid 5,5-dimethyloxazolidine-2,4-dione (DMO) allowed calculation of pHm with the use of values of pHi obtained with BCECF and tubule mitochondrial content. At medium pH (pHe) 7.4, pHi was 7.08 +/- 0.02. Over the pHe range 7.0-7.7, pHi was linearly related to pHe, but the pH gradient across the cell membrane decreased as pHe was lowered. No difference in the relationship between pHe and pHi was obtained when tubules were incubated in the presence of a nonbicarbonate or bicarbonate-buffered medium. Changes in pHe with bicarbonate buffered media resulted in identical pHi values, whether the changes were induced by altered bicarbonate or CO2 content. At pHe 7.4, pHm was 7.78 +/- 0.6 in bicarbonate-buffered medium but was higher (0.2-0.3 pH units) when tubules were bathed in nonbicarbonate-buffered medium. pHm was linearly related to pHi in either buffer. The pH gradient across the inner mitochondrial membrane was also positively correlated with pHe. The present studies indicate the suitability of the techniques for estimating pHi and pHm simultaneously in suspensions of rat renal cortical tubules. Parallel changes occur in both intracellular compartments when pHe is altered. pHm, which is approximately 0.7 pH units greater than pHi, decreases in acute acidosis. This decrease may be important in stimulating renal ammoniagenesis, possibly by activation of alpha-ketoglutarate dehydrogenase. PMID- 2544109 TI - Enalapril decreases angiotensin II receptors in subfornical organ of SHR. AB - We studied brain angiotensin II (ANG II) receptors by quantitative autoradiography in adult normotensive Wistar-Kyoto (WKY) rats and in spontaneously hypertensive rats (SHR) after treating the rats with the converting enzyme inhibitor enalapril, 25 mg/kg, po daily for 14 days. Enalapril treatment decreased blood pressure in only SHR, inhibited plasma angiotensin-converting enzyme activity by 85%, and increased plasma ANG I concentration and renin activity in both WKY and SHR. In the untreated SHR animals, ANG II receptor concentrations were higher in the subfornical organ, the area postrema, the nucleus of the solitary tract, and the inferior olive when compared with the untreated WKY rats. Enalapril treatment produced a large decrease in only subfornical organ ANG II receptors of SHR. The selective reversal of the alteration in subfornical organ ANG II receptors in SHR may indicate a decreased central response to ANG II and may be related to the mode of action of angiotensin-converting enzyme inhibitors in this model. PMID- 2544111 TI - Glucocorticoid inhibition of Na-SO4 transport by chick renal brush-border membranes. AB - To examine the effect of glucocorticoids on sulfate transport by the chick (domestic Gallus gallus) renal tubule we dosed 3-wk-old animals with 60 micrograms dexamethasone/100 g body wt at 24 and 6 h before isolation of renal brush-border (BBM) and basolateral membranes (BLM). Dexamethasone treatment significantly reduced Na-dependent sulfate transport by BBM and had no effect in paired membranes on bicarbonate, proton, or electrical gradient-driven sulfate transport. The glucocorticoid treatment had no statistically significant effect on HCO3-SO4 exchange in the BLM. Kinetic analysis of the dexamethasone effect on the Na-SO4 transport process showed that apparent Vmax was significantly decreased to almost one-half that seen in controls (from 676 to 348 pmol.mg protein-1.5 s-1). The Km in control BBM was 0.40 +/- 0.095 mM and was not significantly different in dexamethasone-treated membranes (0.53 +/- 0.094 mM). To determine whether the dexamethasone-induced decrease in Na-SO4 transport by BBM was indirectly caused by stimulation of Na-H exchange and more rapid dissipation of the initial Na gradient used to drive sulfate uptake, we examined the effect of 0.1 mM amiloride on Na-SO4 uptake by BBM. With amiloride present, dexamethasone treatment caused Vmax to significantly drop from 1,102 to 660 pmol.mg protein-1.5 s-1. Amiloride had no statistically significant effect on the Km. The extent to which amiloride increased Na-SO4 transport and blocked 22Na uptake by BBM did not appear to be related to hormone treatment. The data indicate that glucocorticoids may participate in the regulation of sulfate excretion. PMID- 2544103 TI - Disparate effects of Ca channel blockade on afferent and efferent arteriolar responses to ANG II. AB - Previous reports have suggested that organic calcium antagonists only partially inhibit the renal hemodynamic actions of angiotensin II (ANG II). This study tested the hypothesis that the calcium antagonist-sensitive component of ANG II induced vasoconstriction is localized at a preglomerular site. Videomicroscopic measurements of vascular dimension were performed on in vitro blood-perfused juxtamedullary nephrons from captopril-treated rats. Under control conditions, afferent and efferent arteriolar diameters averaged 23.0 +/- 1.6 and 21.2 +/- 2.2 microns, respectively. Topical application of 0.1 nM ANG II decreased the diameters of afferent (-17 +/- 2%) and efferent (-15 +/- 3%) arterioles. Both 50 microM verapamil and 10 microM diltiazem dilated afferent arterioles. Verapamil also elicited a modest efferent vasodilation. In the presence of either verapamil or diltiazem, the effect of ANG II to decrease efferent diameter was sustained ( 15 +/- 4%); however, the effect of ANG II on afferent diameter was abolished (-1 +/- 1%). These observations document differential influences of calcium channel blockers on ANG II-mediated vasoconstriction and suggest that the pre- and postglomerular vasoconstrictor actions of ANG II may occur through different calcium entry or mobilization mechanisms. PMID- 2544110 TI - Effect of ischemia on sarcolemmal Na+-Ca2+ exchange in neonatal hearts. AB - We studied the effect of cardiac ischemia on sarcolemmal enzymes, Na+-Ca2+ exchange, Ca2+ binding, and Ca2+ efflux in the newborn and adult rabbit. Rabbit ventricle was made ischemic by incubation in hypoxic, glucose-free Tyrode solution at 37 degrees C for 60-120 min. Ischemia inhibited Na+-K+-ATPase and K+ p-nitrophenylphosphatase (PNPPase) activity in the adult myocardium more than in the newborn. In the oxygenated (control) hearts, Na+-Ca2+-exchange activity in the newborn sarcolemma [Michaelis constant (Km) 18 microM; maximum velocity (Vmax) 33] was similar to that in the adult (Km = 16 microM, Vmax = 32). After 60 min ischemia, however, Na+-Ca2+ exchange in the newborn (Km = 16 microM, Vmax = 18) was inhibited less than in the adult (Km = 25 microM, Vmax = 18). In the two age groups, Ca2+ binding and efflux rate were not increased after ischemia, which suggested that Ca2+ permeability did not increase during ischemia. In conclusion, ischemia inhibited sarcolemmal enzymes and Na+-Ca2+ exchange in the newborn less than in the adult, and this lesser inhibition might contribute to or be caused by the greater tolerance of the newborn heart to ischemia. PMID- 2544108 TI - Kinetics of adenine nucleotide catabolism in coronary circulation of rats. AB - We have used the rat isolated, perfused heart to study the metabolism of adenine nucleotides on a single passage through the coronary circulation. Low doses (3-30 nmol) of ATP, ADP, or AMP injected as a bolus were extensively catabolized by ectoenzymes. Increasing doses of each nucleotide demonstrated saturability of catabolism that occurred at significantly lower doses of AMP than of ADP or ATP. The patterns of catabolites formed in each case were consistent with the major pathway of metabolism being sequential dephosphorylation of ATP----ADP----AMP--- adenosine, although from experiments in which [3H]ATP was co-injected with unlabeled ADP, it appears that some direct conversion of ATP----AMP can occur. Furthermore, particularly in the presence of excess unlabeled ATP, [3H]ADP was phosphorylated to [3H]ATP, indicating that ectoenzymes capable of interconverting nucleotides are present. By evaluating recovery and metabolism in serial samples collected rapidly after bolus injection, we were able to use the integrated form of the Michaelis-Menten equation as developed by Bronikowski et al. (Math. Biosci. 61: 237-266, 1982) to derive Michaelis constant (Km) and maximum velocity times capillary plasma volume (Amax) values for adenosinetriphosphatase, adenosine diphosphatase, and 5'-nucleotidase (450, 300, and 93 microM; and 5.3, 5.9, and 1.7 mumol/min, respectively). This analysis also indicated that there is a high degree of heterogeneity of path lengths within the coronary circulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544112 TI - Loose-coupled mitochondria in chronic glucagon-treated hyperthermic ducklings. AB - In chronic glucagon-treated ducklings (GT) showing thermogenic and hyperthermic responses without shivering to glucagon test injection and in control ducklings (TN; both aged 44 +/- 1 days and reared at thermoneutrality), subsarcolemmal (S) and intermyofibrillar (I) mitochondria from gastrocnemius muscle and mitochondria from liver were isolated. Respiration and cytochrome oxidase activity were determined in these isolated mitochondria by polarography and creatine kinase activity by spectrophotometry, both at 25 degrees C. In GT ducklings, the powerful thermogenesis observed in vivo after a glucagon test injection may be due to the uncoupling effect of released free fatty acids (FFA) in loose-coupled mitochondria because their respiration increased as a function of FFA concentration, and the loose coupling of these mitochondria was reversed by addition of albumin. In all types of mitochondria from GT ducklings, the increase in respiration because of FFA was about double that in mitochondria from controls. There was no change in creatine kinase activity from liver and I mitochondria, but a 16% decrease in this enzyme activity (expressed per mg mitochondrial protein) from S mitochondria was shown despite a strong increase in cytochrome oxidase activity from liver mitochondria (+114% if expressed per g tissue) and from muscle mitochondria (I, +53 or +48%; S, +41 or +97% if expressed per mg mitochondrial protein or per g tissue, respectively). These results support a coupling defect in liver and skeletal muscle mitochondria from the GT hyperthermic ducklings and an uncoupling reinforcement by FFA. PMID- 2544115 TI - Well-differentiated cholangiocarcinoma: diagnostic significance of morphologic and immunohistochemical parameters. AB - In order to provide useful criteria for the histologic diagnosis of well differentiated cholangiocarcinoma, we reviewed 62 cases of this type. We assessed the incidences of various cytological as well as structural atypia, positive staining for carcinoembryonic antigen, secretory component, or carbohydrate antigen 19-9. We also assessed alterations in mucin production in comparison with noncancerous bile duct epithelium (48 cases). Using a computer-assisted multivariate analysis, three indicators--including nuclear size variation, formation of a second gland (distended intracytoplasmic lumina or focal cribriform pattern), and positive reaction in carcinoembryonic antigen--were shown to be the most important histologic parameters for the diagnosis of cholangiocarcinoma. One of the three indicators was occasionally positive in noncancerous bile ducts, but two or three indicators were only found in cholangiocarcinoma (p less than 0.001). The majority of the cases of cholangiocarcinoma were indeed positive for two or three of these criteria (54/62; 87%). Among other atypical changes, irregular nuclear configurations, mitosis, and prominent nucleoli showed a high specificity for cholangiocarcinoma. In addition to the major criteria, combined application of these factors can also be helpful for the actual histologic assessment. PMID- 2544116 TI - Tumor-induced osteomalacia and rickets. AB - Tumor-induced osteomalacia is a clinicopathological entity in which vitamin D resistant osteomalacia or rickets occurs in association with a tumor. A total of 72 cases (three current, 69 from review of literature) has been reported to date. Men and women are equally affected. The majority are adults over 30 years old who exhibit progressive lower leg and back pain. Forty bone and 31 soft-tissue tumors were responsible for this syndrome; two-thirds occurred in the extremities. Chemical findings are typical: low serum phosphorus, normal serum calcium, and elevated alkaline phosphatase. Serum levels of 1,25-dihydroxyvitamin D were low or undetectable. Histologically, more than a third were classified as vascular tumors, and half of these cases were hemangiopericytomas that were distributed equally between bone and soft tissues. Other common diagnoses included nonossifying fibromas, "mesenchymal" and giant-cell tumor variants. Features common to all tumors were prominent vascularity, and giant and primitive stromal cells. Only 10 were histologically malignant. Ultrastructural studies have not shown any secretory granules suggestive of a hormone-secreting tumor. It is clear, however, that the tumor is responsible for the osteomalacia because the complete removal generally results in a dramatic reversal of all symptoms and signs. PMID- 2544114 TI - ACTH-induced hypertension in rats: fact or artifact? AB - Evidence from numerous laboratories has shown that administration of adrenocorticotropic hormone (ACTH) to rats produces hypertension within 5 days. However, the analysis of blood pressure in these studies was by the tail-cuff technique, an acute and indirect approach. We have now administered ACTH, via a subcutaneous depot injection (5 or 10 U/day for 9 days), to chronically instrumented rats maintained in metabolic cages. Although tail-cuff measurements of arterial pressure indicated that the ACTH treatment produced hypertension, this was not confirmed by direct 24-h measurements of mean arterial pressure. There was no effect of ACTH on 24-h heart rate throughout the treatment period compared with saline-injected controls. We also examined coefficient of variation of all our measurements. None of the factors was altered by ACTH administration. However, ACTH treatment did produce a diuretic effect, further confirming previous work and providing renal, in addition to cardiovascular, evidence for the bioavailability of the ACTH depot. These results demonstrate that chronic ACTH treatment does not produce a true hypertensive state in rats but rather may enhance the cardiovascular response to the stress of the indirect arterial pressure measurement technique. PMID- 2544113 TI - Glomerular and vascular atrial natriuretic factor receptors in adrenalectomized rats. AB - The renal and vascular responses to atrial natriuretic factor (ANF) and glomerular and vascular ANF receptors were studied in adrenalectomized (ADR) rats with or without deoxycorticosterone (DOC) or dexamethasone (Dexa) replacement therapy. As expected, adrenalectomy elicited hypotension, hemoconcentration, and increased plasma renin activity, but no changes in plasma levels of either ANF-(1 98) or ANF-(99-126) were detected. Dexa treatment decreased both ANF-(1-98) and ANF-(99-126), whereas DOC treatment increased only ANF-(1-98). The acute renal response to ANF and furosemide was reduced in ADR rats and partially restored either by steroid replacement or by raising blood pressure. The blunted natriuretic response to ANF in ADR rats was associated with an increased density of glomerular receptors. Norepinephrine-precontracted vascular strips from ADR rats were more sensitive to ANF (ED50: 1.7 x 10(-8) M) than those from sham operated animals (ED50: 1.5 x 10(-7) M). However, vascular ANF receptor density in mesenteric vessels from ADR animals was decreased. Dexa treatment restored vascular response to that observed in sham-operated animals without a concomitant change in vascular receptor density. Because the presence of guanylate cyclase coupled and noncoupled ANF receptor subtypes have been described in different tissues, we conclude that the apparent lack of correlation between the biological response to ANF and total binding of ANF to glomeruli or mesenteric artery membranes in ADR rats may be in part caused by a differential regulation of both receptor subtype populations. PMID- 2544117 TI - Formoterol fumarate, a new beta 2-adrenoceptor agonist. Acute studies of selectivity and duration of effect after inhaled and oral administration. AB - Four double-blind, double-dummy, randomized, crossover studies were performed in nine asthmatic patients to evaluate beta 2-adrenoceptor selectivity and the duration of effect of formoterol. One study with cumulatively increasing doses of formoterol and salbutamol showed that with the inhaled route, formoterol was 5-15 times more potent than salbutamol with respect to bronchodilation. In a second study, 6 micrograms formoterol and 0.1 mg salbutamol were given for comparison of effect duration. Five hours after salbutamol inhalation the FEV1 values were back to basal. Eight hours after formoterol inhalation about 75% of the maximum bronchodilation remained and the FEV1 was significantly higher than after salbutamol inhalation. By oral route, a study with cumulatively increasing doses showed that as a bronchodilator formoterol was about 50 times more potent than salbutamol. The same potency difference was seen for increase of heart rate and decrease of diastolic blood pressure, indicating that the clinical selectivity for beta 2-adrenoceptors is equal for the two drugs. In a fourth study comparable doses by oral route did not show any difference in the duration of bronchodilation. We conclude that inhaled formoterol is 5-15 times more potent than salbutamol. Inhaled formoterol produces longer duration of bronchodilation, with clinically relevant bronchodilation for at least 8 h. The prolonged duration of formoterol was not seen with oral treatment. PMID- 2544119 TI - [Hormonal changes induced by etomidate in children during the first 24 postoperative hours]. AB - The hormonal effects of an etomidate infusion were assessed postoperatively in children undergoing hypospadias or clubfoot repair. The study was carried out in 12 children, aged between 9 and 70 months, randomly assigned to two equal groups. The anaesthetic protocol was identical for all the children, consisting in a light general anaesthesia (halothane induction, intubation after 60 to 80 micrograms.kg-1 vecuronium) combined with lumbar epidural anaesthesia (initial dose of 0.75 ml.kg-1 bupivacaine with adrenaline, with repeat injections of half the previous dose when there was a change in the haemodynamic parameters suggesting inadequate analgesia). Anaesthetic maintenance was different in both groups: 1 to 1.5 vol% enflurane in a nitrous oxide-oxygen mixture (1/1 v/v) in the control group, and 16.8 +/- 3.0 mg.kg-1.min-1 etomidate infusion in the etomidate group. Venous blood samples were collected after induction of anesthesia (before starting the epidural anaesthesia and the etomidate infusion), at the end of surgery (H0), at the 3rd (H3), 6th (H6), 12th (H12) and 24th h (H24) following surgery. The following hormonal blood concentrations were measured: cortisol, 11 beta-desoxycortisol, aldosterone, 11 beta desoxycorticosterone, dehydroepiandrosterone (DHA) and DHA sulphate (DHA-S). In the control group, cortisol and DHA-S concentrations decreased significantly at H0, aldosterone levels also being significantly lower at H24. In the etomidate group, cortisol concentrations remained significantly lower at H0, H3 and H6; aldosterone concentrations were also significantly lower than control values throughout the study period. There was an important prolonged rise in the concentrations of their precursors. DHA and DHA-S concentrations did not change in the etomidate group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544120 TI - [Primary Sjogren's syndrome complicated by gastric lymphoma. Study of Epstein Barr viral DNA in the tumor]. PMID- 2544121 TI - Stromal regulation of hematopoiesis. PMID- 2544122 TI - Molecular characterization of colony-stimulating factors and their receptors: human interleukin-3. PMID- 2544118 TI - Clinical pharmacology of pipecuronium bromide. AB - The neuromuscular blocking and cardiovascular effects of pipecuronium, in doses ranging 2-3 times its ED95, were evaluated in 46 patients during thiopental, fentanyl, N2O/O2 anesthesia. The neuromuscular blocking effect of pipecuronium was evaluated by recording of the mechanical twitch of the adductor pollicis muscle in response to stimulation of the ulnar nerve at the wrist. Heart rate, systolic and diastolic blood pressures, and cardiac output were non-invasively measured during the onset of the neuromuscular blockade and compared to a saline control group to separate the effect of anesthesia from those of pipecuronium. The mean +/- SD time from administration of pipecuronium to 90% suppression of the first twitch (T1) of the train-of-four was 2.6 +/- 0.8, 2.0 +/- 0.6, and 2.1 +/- 0.6 min following the 70 micrograms/kg, 85 micrograms/kg, and 100 micrograms/kg dose, respectively. There was no significant difference between the different doses of pipecuronium in the time to 90% suppression of T1. In general, all three doses of pipecuronium provided good to excellent intubating conditions within 3 minutes after its administration. The time from the administration of pipecuronium to 5% recovery of T1 was 52.3 +/- 18.2 min in the group given 70 micrograms/kg. This was significantly longer in patients given 85 micrograms/kg (71.9 +/- 15.7 min) or 100 micrograms/kg (71.8 +/- 22.1 min). Times to the start of recovery of T1 and to 25% recovery of T1 showed a similar significant pattern.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544123 TI - The immune system as mediator of virus-associated bone marrow failure: B19 parvovirus and Epstein-Barr virus. PMID- 2544125 TI - The ex vivo correlate of the antithrombotic action of heparin. PMID- 2544124 TI - External messengers and erythropoietin production. AB - We have presented a model for the role of external messenger substances in hypoxic stimulation of kidney production of erythropoietin. These autacoids probably act in concert to activate the adenylate cyclase system to enhance production and/or secretion of erythropoietin. The phosphoproteins generated in this system could act at the level of transcription and translation of erythropoietin as well as at the level of release of erythropoietin from the cell. Even though eicosanoids and beta-2-adrenergic agonists may be involved in mild to moderate hypoxia, it seems more likely that adenosine is more involved in erythropoietin production with increasing severity of hypoxia. Adenosine may play a very early role in hypoxia following the decrease in ATP to trigger erythropoietin production, and hydrogen peroxide may be generated from hypoxanthine, a metabolite of adenosine, during reoxygenation and regional changes in blood flow in the normal kidney and perhaps in certain renal and hepatic tumors. Further work is necessary in vivo to completely clarify the role of adenosine and oxygen free radicals in regulating kidney production of erythropoietin. PMID- 2544126 TI - Effects of unfractionated heparin, dermatan sulfate and low molecular weight heparin on vessel wall permeability in rabbits. PMID- 2544129 TI - Clinical studies with low molecular weight heparins in the prevention and treatment of venous thromboembolism. PMID- 2544128 TI - Biochemical and pharmacologic inequivalence of low molecular weight heparins. AB - LMW heparin fractions obtained from various sources must not be considered as bioequivalent in both the in vitro and in vivo responses. Because of compositional variations, these agents exhibit individual behavior and should be considered as distinct drugs whose safety and efficacy profile must be determined separately. Currently, there is no valid LMW heparin standard available, however, different LMW heparins can be profiled in identical test systems. It is erroneous to assume that most LMW heparins will behave in a similar fashion in terms of safety and efficacy. The need for defined tests to characterized these agents is evident and efforts to profile their actions should be made at both the basic and applied levels. Needless to say, the true efficacy of LMW heparins can only be validated in well-designed randomized clinical trials. Optimization of LMW heparins in preclinical pharmacologic studies, as reported here, is a crucial factor in the development of these agents. The superior clinical efficacy/safety performance of some of the LMW heparins in contrast to other LMW heparins is a result of extensive preclinical pharmacologic investigations undertaken to optimize the therapeutic index of these agents. Such optimization studies have not been conducted during the development of many LMW heparins, resulting in decreased efficacy and undue complications. PMID- 2544130 TI - A monocyte-derived B cell growth factor is IFN-beta 2/BSF-2/IL-6. PMID- 2544131 TI - Characterization of an IL-6-responsive element (IL6RE) present on liver-specific genes and identification of the cognate IL-6-dependent DNA-binding protein (IL6DBP). PMID- 2544127 TI - Pharmacokinetics of heparin and related polysaccharides. AB - The pharmacodynamic profile of standard heparin (SH), a low molecular weight derivative (CY 216) and of dermatan sulfate (DS), a new potential antithrombotic drug, was investigated in the rabbit over a large range of doses. After bolus i.v. injection of low doses, the biological activity of SH disappeared exponentially; however, its half-life was prolonged when the dose injected increased, and over 158 micrograms/kg (100 anti-factor Xa U/kg) the biological activity disappeared as a concave-convex curve. CY 216 disappeared more slowly than SH at low doses but faster than SH at higher doses. More than 90% of the DS biological activity present 1 minute after the i.v. injection disappeared exponentially without dose-dependent effects. Increasing doses of the three drugs were then delivered for 5 h under continuous infusions. Below 500 micrograms/kg/h the DS and CY 216 plateau concentrations were higher than that of SH while above this dose the SH concentration was higher than that of DS and CY 216. These observations may be explained by the results of pharmacokinetics experiments where 125I-labeled compounds were delivered by bolus i.v. injection in association with increasing doses of their unlabeled counterparts. For SH there was a 10-fold difference between the half-life of the lower dose (32 micrograms/kg or 5 anti-factor Xa U/kg) and that of the higher dose (3200 micrograms/kg); it was demonstrated that the half-life of SH continuously shortened as its plasma concentration decreased. In contrast the CY 216 and DS half-lives were very close, independent of the dose delivered, and therefore longer than that of SH at low doses and shorter than that of SH at higher doses. The renal contribution to the clearance of SH, CY 216, and DS was also investigated by injecting a low dose (150-200 micrograms/kg) and a 10 times higher dose to sham-operated or binephrectomized animals: renal function is critical for the elimination of the three compounds from the blood except for SH at low doses. These observations are promising for the development of DS and may have clinical implications. PMID- 2544132 TI - [Enalapril versus digoxin in chronic congestive cardiac failure. Study of variation of the left ventricular ejection fraction determined by radio-isotopic angioscintigraphy at rest and during exertion]. AB - The effects of enalapril were evaluated in a double-blind, controlled study during 8 weeks, in patients with stable, congestive chronic cardiac insufficiency (functional classification II and III of the NYHA), in sinus rhythm, treated with digitalis and diuretics. 20 patients were randomly divided in two groups: one group continuing the digoxin (group A: 10 patients) and one group where enalapril was substituted for digoxin (group B: 10 patients). Patients from group B presented less clinical aggravation during the study. The left ventricular stroke volume (SV) is significantly decreased in group B at rest (0.21 +/- 0.06 at 50; 0.18 +/- 0.04 at 54; p less than 0.05), while it remained stable during stress. No variations of the SVs were noted at rest and during stress in group A. Considering its favorable clinical effects and after evaluation of its longterm side effects, enalapril may be an acceptable alternative to digitalis in cardiac insufficiency with sinus rhythm, except in patients for whom a drop in the systemic blood pressure or an increased kaliemia or creatininemia, could be potentially harmful. PMID- 2544135 TI - Assessment of the potency of different standards in the immunoradiometric assay of ACTH. AB - There is no generally accepted human international reference preparation for ACTH. The different centres offering clinical ACTH measurement therefore select assay standards from the limited range of human-purified and synthetic preparations currently available. We have examined the relative potencies of three synthetic ACTH standards in comparison with the NIBSC human-purified 1-39 (code 74/555) in a two-site immunoradiometric assay (IRMA) for ACTH. All of the standards produced parallel curves in the IRMA but there was a wide variation in potency between the preparations giving highly significant differences in quality control and patients' ACTH results. Standards supplied by the National Hormone and Pituitary Programme, Bachem UK Ltd and Universal Biologicals Ltd gave lower ACTH results than results calculated using the NIBSC preparation. The potency differences between these standards emphasise the need for a reference preparation and, in the meantime, the necessity to define normal reference ranges for each laboratory. PMID- 2544134 TI - Serum angiotensin converting enzyme in sarcoidosis--its value in present clinical practice. AB - Angiotensin converting enzyme (ACE) has a central role in blood pressure homeostasis. It is present in low and fairly constant concentration in the serum and in high concentration in the pulmonary capillary bed. Unusually high serum angiotensin converting enzyme (SACE) activity is present in active sarcoidosis, an observation now confirmed by many investigators. In spite of its lack of specificity as a test for sarcoidosis, SACE provides a good monitor of disease activity which clinicians find useful in the management of patients with sarcoidosis. There continues to be considerable interest in SACE in sarcoidosis and with the recent development of simpler assays, more centres may be expected to offer SACE measurements as a service. In this paper we discuss the indications for estimating SACE in sarcoidosis and its relevance to current clinical practice. PMID- 2544133 TI - [Echography and scintigraphy using technetium 99m pyrophosphate in the diagnosis of cardiac amyloidosis]. AB - The diagnosis of amyloid cardiomyopathy was only based, until the last few years, on the results of invasive techniques. It seems presently that the combined contribution of cardiac sonography and scintigraphy using technetium 99m pyrophosphate, makes, most of the time, this diagnosis possible without need for additional examinations. This notion is illustrated by a typical case-report and data from the literature. Demonstration on the cardiac sonogram of a thickening of the walls-while the context and especially the electrocardiogram are not in favor of a left ventricular hypertrophy--associated with a very particular "hyperechoing" aspect and an abnormal fixation on the scintigram, may be considered specific of this disease. PMID- 2544136 TI - Pathogenesis and treatment of human genital papillomavirus infections: a review. AB - HPV infection of the genital tract is common and anogenital warts or condyloma acuminatum is increasing rapidly in incidence. In addition, certain HPV types are closely associated with genital tract malignancies. Although recent advances in molecular biology have led to an increased understanding of the organization and functions of the papillomavirus genome, the pathogenesis of HPV infections and host responses to these diseases remain poorly understood. Treatment of anogenital warts is difficult and no completely satisfactory treatment modality is currently available. Comparatively few therapeutic modalities have been thoroughly evaluated, although recent studies of intralesionally and parenterally administered interferons have demonstrated beneficial effects of interferon compared to placebo. Additional studies of treatment for condyloma acuminatum are needed and should include the use of biologic response modifiers such as interferons, as well as antiviral drugs, with or without conventional methods of local therapy. PMID- 2544142 TI - The reactions of horseradish peroxidase, lactoperoxidase, and myeloperoxidase with enzymatically generated superoxide. AB - The formation and decay of intermediate compounds of horseradish peroxidase, lactoperoxidase, and myeloperoxidase formed in the presence of the superoxide/hydrogen peroxide-generating xanthine/xanthine oxidase system has been studied by observation of spectral changes in both the Soret and visible spectral regions and both on millisecond and second time scales. It is tentatively concluded that in all cases compound III is formed in a two-step reaction of native enzyme with superoxide. The presence of superoxide dismutase completely inhibited compound III formation; the presence of catalase had no effect on the process. Spectral data which indicate differences in the decay of horseradish peroxidase compound III back to the native state in comparison with compounds III of lactoperoxidase and myeloperoxidase are also presented. PMID- 2544137 TI - Treatment of FeLV-induced immunodeficiency syndrome (FeLV-FAIDS) with controlled release capsular implantation of 2',3'-dideoxycytidine. AB - 2',3'-dideoxycytidine (ddC) inhibits replication of the immunodeficiency inducing strain of feline leukemia virus (FeLV-FAIDS) in vitro at concentrations ranging from 1-10 micrograms/ml. Additive antiviral effect is achieved when ddC is combined with either human recombinant alpha interferon (IFN alpha) or tumor necrosis factor (TNF) plus IFN alpha. Initial in vivo pharmacokinetic studies in cats, utilizing bolus intravenous administration of ddC (20 mg/kg), resulted in peak plasma concentrations of 15 micrograms/ml 1 min after administration and a half-life of approximately 1 h. These values could not be augmented with high levels of the deaminase blocker tetrahydrouridine administered prior to or concurrently with ddC. In vivo trials utilizing multiple, daily intravenous injections of ddC could not prevent the development of persistent viremia in cats infected with FeLV-FAIDS. To enhance ddC pharmacokinetics and antiviral activity, controlled release capsular implants were developed by blending ddC with a copolymer consisting of DL-lactide glycolide and hydroxypropyl cellulose, which was melt-spun into fibers and encapsulated in a sheath of polyethylene glycol for subcutaneous implantation. Pharmacokinetic studies, conducted in cats receiving an average dose of 600 mg of ddC, indicated an average peak plasma concentration of 17 micrograms/ml achieved at 6 h post implantation with 3.5 micrograms/ml noted at 48 h; and an extension of plasma half-life from 1.5 (bolus subcutaneous injection) to 20 h. sustained plasma concentrations of 1.5 to 10 micrograms/ml, equivalent to ddC levels previously shown to have anti-FeLV activity in vitro, were maintained throughout a 72 h period. Implantation devices could be replenished every 48 h and elevated plasma levels were sustained for four weeks without signs of clinical toxicity, sepsis or significant alterations in the hemogram. Initial clinical trials employing controlled release capsular ddC implants in vivo indicate significant retardation of FeLV-FAIDS replication throughout a four week treatment period. PMID- 2544141 TI - Distribution of endoplasmic reticulum and calciosome markers in membrane fractions isolated from different regions of the canine brain. AB - Four regions of the canine brain (frontal lobe, parieto-occipital lobe, brainstem, and cerebellum) were each fractionated by differential centrifugation into a crude mitochondrial pellet (P2) and a crude microsomal pellet (P3). Markers of endoplasmic reticulum (glucose-6-phosphate phosphatase and rotenone insensitive NADPH cytochrome c reductase) and markers of the 1,4,5-trisphosphate (IP3)-sensitive Ca2+ store ([3H]IP3 binding and IP3-induced Ca2+ release) were measured. No correlation was found between the two classes of markers, which suggests that the IP3 receptor does not belong to the endoplasmic reticulum in canine brain. Cerebellum P2 and P3 fractions displayed levels of [3H]IP3 binding 10- to 30-fold higher, and rates of IP3-induced Ca2+ release greater than 15-fold faster than the homologous cerebrum and brainstem fractions. Actively accumulated Ca2+ was only partially released by IP3, both before and after saponin disruption of the plasma membrane compartment. The proportion of the IP3-sensitive Ca2+ store relative to that of the total (IP3-sensitive and IP3-insensitive) Ca2+ store was variable; i.e., it was larger in cerebellum P2 (approximately 90%) than in cerebrum fractions (less than 30%). Cerebellum fractions constitute the best source from which an IP3-sensitive Ca2+ storing organelle can be purified. PMID- 2544144 TI - Polyclonal antibodies to rabbit skeletal muscle protein phosphatases C-I and C II. AB - Polyclonal antibodies against rabbit skeletal muscle phosphatases C-I and C-II were raised in goats and in mice. The goat polyclonal antibodies to phosphatases C-I and C-II were examined for their ability to immunoblot the purified enzymes and crude rabbit muscle extracts. In preparations of phosphatases C-I and C-II that were apparently homogeneous, the expected ca. 35- to 38-kDa polypeptides were immunoblotted, but, in addition, immunoblotting of a 67-kDa polypeptide was observed. Both the antisera blotted only the 67-kDa polypeptide in crude rabbit muscle extracts and not the expected 35- to 38-kDa polypeptides. These findings are qualitatively similar to those reported previously (D.L. Brautigan et al. (1985) J. Biol. Chem. 260, 4295-4305) where immunoblotting experiments with a sheep antisera to phosphatase C-I indicated that the ca. 35-kDa polypeptide originates from a 70-kDa precursor. On further investigation, it was found that our antisera were strongly immunoreactive to rabbit serum albumin. The antisera blotted purified rabbit albumin, but not bovine serum albumin. After passage through a rabbit albumin-Sepharose column, the antisera lost immunoreactivity to rabbit albumin, and no longer blotted the ca. 70-kDa band in muscle extracts or in purified enzyme preparations. These findings show that the phosphatase preparations contained traces of albumin which produced a strong antigenic reaction. Production of antisera in BALB/c mice produced similar results; i.e., an antibody to the low-molecular-weight phosphatases was produced that was also a strong antibody to rabbit albumin. This antibody could be removed by affinity adsoption on rabbit albumin-Sepharose columns. In addition, the antibodies to phosphatase C-I displayed no cross-reactivity to phosphatase C-II, while antibodies to C-II showed no cross-reactivity to phosphatase C-I by immunoblotting methods. PMID- 2544145 TI - Changes in membrane constituents and chemiluminescence in vitamin E-deficient red blood cells induced by the xanthine oxidase reaction. AB - The oxidation of vitamin E-deficient rat red blood cells (RBCs) induced by the hypoxanthine-xanthine oxidase (HX-XOD) system has been performed in an aqueous suspension. The generation of chemiluminescence and the accumulation of thiobarbituric acid-reactive substances (TBARS) were observed initially and were followed by hemolysis. Interestingly, the total counts of chemiluminescence were closely related to the amount of TBARS. The predominant change of membrane proteins induced by the reaction was the depletion of spectrin bands in gel electrophoresis. When RBC ghosts were oxidized with HX-XOD, the sulfhydryl (SH) groups of membrane proteins decreased at an early stage of the incubation, which was coincident with the above protein alteration. Membrane alpha-tocopherol suppressed not only the formation of TBARS but also chemiluminescence and hemolysis; nevertheless, it did not inhibit the protein damage and the loss of SH groups. Moreover, it was concluded that the chemiluminescence observed during the oxidation of RBC membranes was associated mainly with the peroxidation of lipids and only to a minor extent with the oxidation of proteins. PMID- 2544138 TI - Effects of denaturants at low concentrations on the reversible denaturation of staphylococcal nuclease. AB - Three very unstable mutant forms of staphylococcal nuclease were used to quantitate the change in the apparent equilibrium constant for reversible denaturation (Kapp) as a function of denaturant concentration for a variety of different denaturing solutes. The value of this equilibrium constant in the absence of denaturant (Kapp,0) was determined by renaturation of the mutant proteins with a combination of glycerol and calcium ion, the latter of which binds at the active site in the native conformation. Because Kapp,0 fell in the easily measurable range between 0.1 and 1, the change in Kapp, and thus the change in free energy (delta Gapp), at very low concentrations of denaturants could be accurately measured. With guanidine hydrochloride (GuHCl), the rate of change of the apparent free energy of denaturation with respect to denaturant concentration (d(delta Gapp)/dCGuHCl or mGuHCl) was found to be remarkably constant down to zero denaturant concentration, even though this value was different for each of the three proteins. Unlike GuHCl, urea exhibited a slightly reduced value of d delta Gapp/dCurea at low concentrations. Results with a number of thiocyanate, perchlorate, and iodide salts confirmed that the Hofmeister series holds for concentrations below 0.1 M; that is, with regard to efficacy as a denaturant SCN- greater than ClO4- greater than I- and Li+,NH4+ greater than Na+,K+. However, all of the chaotropic salts analyzed exhibited markedly increased values of d(delta Gapp)/dCsalt at concentrations below 0.2 M. One possible explanation for these large deviations from a linear relationship between delta Gapp and salt concentration is that weak binding or adsorption of chaotropic anions is occurring at a saturable number of sites in hydrophobic regions of the denatured state. PMID- 2544139 TI - Murine erythroleukemia cells possess an active ubiquitin- and ATP-dependent proteolytic pathway. AB - The ubiquitin (Ub)-dependent proteolytic pathway may function in selective elimination of cellular proteins during erythroid differentiation. Murine erythroleukemia (MEL) cells, which can be induced to differentiate to reticulocytes in culture, may provide a convenient system for studying the role of Ub-dependent proteolysis in erythroid differentiation. The following observations indicate that MEL cells possess an active Ub-dependent proteolytic pathway. (i) Addition of purified Ub to MEL cell fraction II (Ub-depleted lysate) stimulated ATP-dependent degradation of radioiodinated proteins. (ii) Covalent conjugation of carboxyl termini of Ub molecules to substrate protein amino groups is a necessary step in Ub-dependent degradation. Des-glygly-Ub (Ub lacking its carboxyl-terminal glygly moiety) did not stimulate protein degradation in MEL cell fraction II. (iii) The Ub-dependent component of protein degradation in MEL cell fraction II was specifically inhibited by amino acid derivatives that are inhibitors of Ub-protein ligase. (iv) MEL cell fraction II contained apparent homologs of all of the rabbit reticulocyte Ub carrier proteins (E2's) except E2(20K) and E2(230K). Ub-dependent proteolysis was seen only in MEL cell lysates prepared in the presence of leupeptin; an enzyme of the proteolytic pathway was inactivated if leupeptin was omitted. PMID- 2544143 TI - In vitro kinetics of reduction of cytochrome c554 isolated from the reaction center of the green phototrophic bacterium, Chloroflexus aurantiacus. AB - The photochemical reaction center in the green bacterium Chloroflexus aurantiacus is similar to that found in purple phototrophic bacteria and interacts with a multiheme membrane-bound cytochrome. We have examined the kinetics of reduction of the pure solubilized reaction center cytochrome by laser flash photolysis of solutions containing lumiflavin or FMN. Reduction by lumiflavin semiquinone followed single exponential kinetics and the observed rate constant (kobs) was linearly dependent on protein concentration (k = 1.8 X 10(7) M-1s-1 heme-1). This result suggests either that the four hemes have similar reduction rate constants which cannot be resolved or that there are large differences in rate constant and only the most reactive heme (or hemes) was observed under these conditions. To determine the relative reactivities of the four hemes, we varied the extent of heme reduction at a single total protein concentration. As the hemes were progressively reduced by steady-state illumination prior to laser flash photolysis, kobs for the reaction with fully reduced lumiflavin decreased nonlinearly. Second-order rate constants for the four hemes were assigned by nonlinear least-squares analysis of kobs vs oxidized heme concentration data. The second-order rate constants obtained in this way for the highest and lowest potential hemes differed by a factor of about 20, which is larger than expected for c-type cytochromes based on redox potential alone (a factor of about 3 would be expected). This is interpreted as being due to differences in steric accessibility. Relative to the highest potential heme, which is as reactive as a typical c-type cytochrome, we estimated a steric effect of approximately twofold for heme 2, and steric effects of approximately fivefold for hemes 3 and 4. Using fully reduced FMN as reductant of oxidized cytochrome, ionic strength effects indicate a minus-minus interaction, with approximately a -2 charge near the site of reduction of the highest potential heme. PMID- 2544146 TI - [Oncogenes in human gastric carcinoma]. AB - Alteration of oncogene and loss of chromosomal heterozygosity are infrequent in human gastric carcinoma compared with those in other gastrointestinal carcinomas. Amplification of c-erbB-2 gene is observed in well differentiated adenocarcinoma, while sam gene is found in poorly differentiated adenocarcinoma or scirrhous carcinoma. sam gene, which was isolated from a gastric cancer cell line KATO-III by a DNA renaturation method, encodes tyrosine-specific protein kinase domain. A good correlation evidently exists between the synchronous expression of TGF alpha and ras p21 and biological malignancy of gastric carcinoma. c-myc and c-fos proteins are found not only in tumor cells but also in stromal cells including macrophages and fibroblast around the tumors. The prognosis of patients with c myc p 62-positive stromal cells is significantly better than that of patient with p 62-negative stromal cells. Coamplification of the hst-1 gene and int-2 is observed in 50% of primary tumors and all metastatic tumors of esophageal carcinoma. PCR (polymerase chain reaction) technique seems to be useful for the detection of oncogene point mutation in human gastric carcinoma. PMID- 2544140 TI - Purification and characterization of the sesquiterpene cyclase aristolochene synthase from Penicillium roqueforti. AB - The sesquiterpene cyclase, aristolochene synthase, has been purified from Penicillium roqueforti by gel filtration and anion-exchange chromatography. Isolation was facilitated by a change in the elution behavior of the enzyme during gel filtration at different steps in the purification. The purified enzyme had a specific activity of 70 nmol/min/mg protein. The molecular weight as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was Mr 37,000. The native molecular weight as determined by gel filtration chromatography was Mr 48,000. The requirement for Mg2+ could be partially substituted with 0.01 mM Mn2+, but higher concentrations were inhibitory. Pyrophosphate, a competitive inhibitor of most terpene cyclases, had no effect on enzyme activity up to a concentration of 5.0 mM. The maximum activity was observed between pH 6.25 and pH 7.50, and the Km for farnesyl pyrophosphate was 0.55 +/- 0.06 microM. PMID- 2544148 TI - [Antitumor activities of oily suspended YM881 (SMANCS) against VX2 carcinoma]. AB - YM881 (MW 15,000) is a conjugate protein preparation of two copolymer of styrene maleic acid [SMA] (MW 1,500) and neocarzinostatin [NCS] (MW 12,000). Antitumor activities of YM881 and NCS with or without oily solvent injected arterially against VX2 carcinoma of rabbits were examined. Based on the growth and histological examination of the tumor, remarkable anti-tumor activities were observed with oily suspended YM881 but not with the other group. This results indicate that targeting of the anticancer agent to the tumor is most important for eliciting of antitumor activities. PMID- 2544151 TI - [Peripheral neuropathy caused by cisplatin in patients with lung cancer]. AB - The correlation between peripheral neuropathy and cisplatin (CDDP) was elucidated in 27 patients with primary and metastatic lung cancer, who were treated with Adriamycin 30 mg/m2 day 1, CDDP 80 mg/m2 day 1, and VP-16 70 mg/m2 day 1-5 every 4 weeks. The incidence of peripheral neuropathy was 33% (9 of 27 patients) and it increased to 60% in the patients who received over 320 mg/m2 of CDDP, demonstrating a positive correlation between the incidence of this toxicity and the total dose of CDDP. However, no significant relation was observed between the grade of neuropathy and CDDP. The neuropathy was manifested in the sensory system of the distal extremities and was developed into proximal portions. The peripheral neuropathy with grade 3 was irreversible, resulting in the dose limiting factor of this regimen. PMID- 2544147 TI - [Characteristics in antitumor effects of organic silicon related compounds]. AB - To investigate the characteristics in antitumor effects of 2 trimethylsilylethylthioethylamine(KAS-010) and its conjugate with 5-FU (KAS-011), the antitumor and immunomodulating activities of these silicon compounds were examined with various systems. Both KAS-010 and KAS-011 administered orally was found to be effective to B 16 melanoma, Meth A sarcoma and MM 46 mammary carcinoma in vivo. On the other hand, KAS-011 administered orally exhibited a marked antitumor activity against L 1210 leukemia bearing mice. Furthermore, these silicon compounds inhibited significantly metastases to the lymph nodes and lung of Lewis lung carcinoma implanted id into the right ear of BDF1 mice. Especially, KAS-011 in combination with tumor amputation resulted in a remarkable prolongation of the survival time (% ILS: 93.8%) in this antimetastatic model. The cell killing effect was mainly dependent on the exposure time of these silicon compounds in cultured KB and human lung cancer (OAT) cells. Moreover, a significant increase of delayed type hypersensitivity reaction (DTHR) to sheep red blood cell (SRBC) induced by KAS-010 was seen in old aged mice. The DTHR in B 16 melanoma and Ehrlich carcinoma bearing mice treated with KAS-010 was significantly higher than those of non-treated tumor bearing mice, indicating an enhanced cellular immunity to KAS-010 possibly resulting in a remarked antitumor effect. We also found that tumor free mice treated these silicon compounds were acquired specific tumor immunity to Meth A sarcoma and MM 46 mammary carcinoma. PMID- 2544150 TI - [Adriamycin, cisplatin, and etoposide combination chemotherapy for small cell lung cancer]. AB - Twenty-three patients with small cell lung cancer (11 with limited disease and 12 with extensive disease) who had not received previous chemotherapy were treated with a combination of adriamycin (30 mg/m2, i.v., on day 1), cisplatin (80 mg/m2, i.v., on day 1) and etoposide (70 mg/m2, i.v., on day 1-5). This chemotherapy regimen was repeated at 3- or 4-week intervals for 3 to 5 treatment cycles. Among 22 evaluable patients, 5 showed complete response and 17 had a partial response (response rate 100%). The median response duration of 12 extensive disease patients was 21 months. There were 5 survivors for more than 2 years. Toxicity included moderate to severe hematologic toxicity, alopecia, nausea and vomiting. This combination chemotherapy appears to be optimal for the treatment of small cell lung cancer. PMID- 2544149 TI - [Pilot phase II study of hybrid chemotherapy of CAV-PVP in small cell lung cancer (SCLC)]. AB - A pilot phase II study of a hybrid chemotherapy for SCLC has been conducted between October 1986 and March 1988. Dose and schedule of the regimen were as follows: CTX, 700 mg/m2, on day 1; ADM 30 mg/m2, on day 1; VCR, 1.4 mg/m2, on day 1 (CAV); and CDDP, 60 mg/m2, on day 8; VP-16, 100 mg/m2, on days 8 and 9 (PVP). Courses were repeated q. 4 weeks up to 6 cycles. Patients with LD received chest irradiation at a dose of 50 Gy when maximal response was achieved. Thirty-six patients were fully evaluated for tumor response and toxicity. All 18 patients with LD responded to the regimen including 11 CRs (61%); there were 7 CRs (39%) and 9 PRs (50%) in patients with ED. Fourteen of the 18 patients with LD have survived for 7 to 22 months, against 12.8 months in ED patients. The major toxicity was myelosuppression, but it was well tolerated. These results indicate that hybrid chemotherapy is highly effective for SCLC, and warrants further clinical trials. PMID- 2544152 TI - A prospective randomized trial of low molecular weight heparin-DHE and conventional heparin-DHE (with acenocoumarol) in patients undergoing gynaecological surgery. AB - The antithromboembolic efficacy of once a day low molecular weight heparin in fixed combination with dihydroergotamine (LMWH-DHE) was compared with conventional heparin-DHE in combination with Acenocoumarol (heparin-DHE/A) in 191 patients undergoing gynaecological surgery. LMWH-DHE proved equally effective in preventing thromboembolic complications, with a similar incidence of postoperative bleeding and side effects. Deep vein thrombosis occurred once in each group and one non-fatal pulmonary embolism occurred in the LMWH-DHE group. The main advantage of LMWH-DHE was significantly better patient acceptance of the single daily subcutaneous injection as compared with the two injections of conventional heparin-DHE (P = 0.02). On the other hand, LMWH-DHE was associated with significantly increased incidence of intraoperative bleeding (P less than 0.02). The bleeding did not, however, cause any clinical problems. Discontinuation of therapy due to bleeding or pain at the site of injection occurred three times in each group. We consider the use of LMWH-DHE to be an attractive, economic and safe method of thromboembolic prophylaxis. PMID- 2544153 TI - [Parathormone, cyclic AMP, 1,25 dihydroxyvitamin D and osteocalcin in hypercalciuric renal lithiasis]. AB - A study was undertaken in 46 subjects; 21 patients diagnosed as having HRL and 25 volunteers patients. Biochemical and hormonal analyses were performed in the study population, including determination of Ca, P, Mg, Cr in blood and urine, phosphate tubular resorption (PTR), maximum tubular phosphate resorption (MTPR), fasting calcium secretion (FCS), alkaline phosphatase (AP), hydroxyprolinuria (HPR), osteocalcin (BGP), parathormone (PTH), cAMP, and 1-25(OH)2D. The stone formers showed lower calcemia values (p less than or equal to 0.005d), higher phosphaturia, and magnesiuria (p less than or equal to 0.0005), higher FCS (P less than or equal to 0.005) and higher values for PTH (p less than or equal to 0.01) and cAMP (p less than or equal to 0.0025). No significant differences were observed for the other parameters evaluated. Classification of the patient group into 2 subgroups (renal SbR and absorptive SbA) according to FCS values greater or lower that 0.16 mg/dl, the SbR patient group revealed a higher PTH and 1 25(OH)2D values (p less than or equal to 0.05). There appears to be no increase of bone resorption since AP, HPR, and BGP values in our patients fell within normal ranges. The 1-25(OH)2D levels were also normal and, with respect to the control group, were only elevated for the SbR patient group, whose PTH levels were also observed to be elevated. These increments appear to be related and may result in intermediate forms between renal and absorptive hypercalciuria. PMID- 2544154 TI - [Wilms' tumor associated with membranous glomerulonephritis]. AB - We report on a case of Wilms tumor associated with segmental, focal membranous glomerulonephritis in the contralateral kidney of a 4-year-old boy. The possible etiopathogenesis and outcome of this rare disease entity are discussed. Except for rare cases, this condition usually progresses to end-stage renal failure. PMID- 2544156 TI - [The dynamics of the content of ammonia and volatile fatty acids in the rumen fluid during the use of dry feed rations in the cow]. AB - The development of the pH-value, the molar quota of volatile fatty acids and of the NH3 content in the change from silage rations to dried rations on the basis of pelleted feed and in dependence on the time after the beginning of feeding was tested in studies of the rumen fluid of four cows in order to explain the causes of the very low NH3 content of the rumen fluid in feeding experiments with rations based on pelleted straw-concentrate mixtures. An influence of the ration type and the time after the beginning of feeding on the molar quota of the individual volatile fatty acids and the NH3 content was ascertained. The ration type tested can cause an NH3 content in the rumen fluid below 1 mmol/l. PMID- 2544160 TI - Cervical atypia. AB - Abnormal Papanicolaou smears are common. The incidence of cervical intraepithelial neoplasia in young women is increasing. These events trigger a strong emotional reaction in a woman and create stress in her relationships. The availability of accurate information and appropriate counselling will assist a woman and her partner to deal with this event in their lives. PMID- 2544159 TI - Prevalence of cytomegalovirus cervical excretion in pregnant women in Hong Kong. AB - In 1986 a study was made on the prevalence of cervical colonization by cytomegalovirus (CMV) in normal third trimester pregnant Chinese women in Hong Kong. Seven out of 95 unselected subjects had positive CMV culture, making a prevalence of 7.4%. This is a low prevalence when compared with the prevalence found in other people of Asian origin and similar to the prevalence found in the Caucasians. PMID- 2544158 TI - Gonadal dysgenesis: a review of 15 years' management. AB - Fifteen years' experience in a Menstrual Disorder Clinic revealed that gonadal dysgenesis was the commonest cause of primary amenorrhoea. Investigations using Barr Body study and laparoscopic examination were unsatisfactory and inaccurate. The invasive nature of laparoscopy was the likely principal cause for the high default rate in the early years of the Clinic. The availability of hormone radioimmunoassay and cytogenetic study has improved the diagnostic acuity. Gonadal malignancy associated with the "Y" chromosome could also be identified at its early stage. Oestrogen replacement therapy has been found to be beneficial and is associated with little side effects and no endometrial pathology has been identified. PMID- 2544157 TI - [The energy maintenance requirement of growing pigs of different sexes give normal and high protein diets. 2. Experiments in sows]. AB - Six animals each out of 12 female fattening hybrids (150 X (L X E] were given rations containing 17 or 45% crude protein resp. 4 periods of growth feeding alternated with 5 periods of maintenance feeding in the live weight range between 33 and 146 kg. The feeding level did not have an influence on the level of digestibility. From the experiments with 17% crude protein in the ration 1004 and from those with 45% crude protein 947, on average 977 kJ metabolizable energy per kg LW0.62 in the maintenance periods and 980 in the growth periods were derived as maintenance requirement of metabolizable energy. These values for maintenance requirement are by 50% higher than those in previous experiments of our work team. In contrast to expectations, energy maintenance requirement did not grow due to the increase of the protein content of the rations. The utilization of metabolizable energy for retention was 73% for rations with a normal protein content and 66% for those with a high protein content. The partial utilization of metabolizable energy for fat retention was ascertained as 83% and as 49% for protein retention by means of multiply regressive evaluation. PMID- 2544161 TI - Two pathways for phosphate/water oxygen exchange by yeast inorganic pyrophosphatase. AB - A scheme of Mg2+ and Pi binding to yeast inorganic pyrophosphatase has been deduced from the concentration dependencies of the rate of oxygen exchange between Pi and water. The exchange reaction requires the binding of MgPi and free Pi (pathway I) or two MgPi (pathway II) in addition to two Mg2+ ions bound in the absence of Pi. Pathway II predominates above 0.16 mM Mg2+. The rate of formation of bound PPi from bound Pi for pathway II is three times as high as that for pathway I. The results suggest that the binding of the fourth Mg2+ ion to pyrophosphatase stimulates its synthetic vs its hydrolytic capability. PMID- 2544155 TI - Augmented pituitary corticotropin response to a threshold dosage of human corticotropin-releasing hormone in depressives pretreated with metyrapone. AB - We studied pituitary corticotropin response to exogenous corticotropin-releasing hormone infusion and attempted to control for the confounding effect of variable serum cortisol levels between depressed and control subjects. If metyrapone was given during the time of day when hypothalamic pituitary adrenal activity was otherwise low, the relative increase in the corticotropin concentration was small. Pituitary response to exogenous corticotropin-releasing hormone can be defined under conditions in which the amount of glucocorticoid-mediated negative feedback present at the level of the pituitary gland is equal in all subjects. When the ambient cortisol level was equalized (and suppressed) in all subjects at the time of study with a threshold dosage of corticotropin-releasing hormone, we found an augmented response to corticotropin-releasing hormone in depressives. This raises the possibility that either increased pituitary sensitivity to corticotropin-releasing hormone or an increased intracellular pool of corticotropin is available for release in subjects with major depressive illness. PMID- 2544162 TI - Mobilization of intracellular calcium by endothelin in Swiss 3T3 cells. AB - When intracellular free Ca2+ concentration [( Ca2+]i) was monitored in fura2 loaded Swiss 3T3 cells, endothelin increased [Ca2+]i in a dose-dependent manner; after the addition of endothelin, an initial transient peak was observed immediately and was followed by a sustained increase in [Ca2+]i lasting at least 5 min. 45Ca2+ efflux and influx experiments in endothelin-stimulated Swiss 3T3 cells revealed that the change in [Ca2+]i could be explained by a dual mechanism; an initial transient peak induced mainly by the release of Ca2+ from intracellular stores and the sustained increase by an influx of extracellular Ca2+. Cellular generation of inositol 1,4,5-trisphosphate and cyclic AMP were not induced by endothelin, suggesting that other cellular mediators with the capacity to release Ca2+ from intracellular stores play a significant role in the signal transduction pathway of endothelin in Swiss 3T3 cells. PMID- 2544165 TI - Insulin-like growth factor I (IGF-I) production and the presence of IGF-I receptors in rat medullary thyroid carcinoma cell line 6-23 (clone 6). AB - To clarify whether insulin-like growth factor I (IGF-I) is an autocrine growth factor of rat medullary thyroid carcinoma (MTC) cell line, 6-23 (clone 6), IGF-I binding to MTC cell membranes, IGF-I levels in the conditioned culture medium of MTC cells and the effects of IGF-I on methyl-[3H]thymidine incorporation to MTC cells were examined. Scatchard analysis of saturation binding studies revealed the association constant and the maximal binding capacity were 1.0 x 10(9) M-1 and 199 fmol/mg of membrane protein, respectively. The binding of [125I]IGF-I to MTC cell membranes was inhibited by unlabeled IGF-I, IGF-II and insulin; the relative potencies were IGF-I greater than IGF-II much greater than insulin, suggesting the presence of type I IGF receptors in MTC cells. IGF-I levels in the conditioned culture medium of MTC cells were 120 +/- 3 pM (mean + SE). IGF-I (10( 10) to 10(-8) M) dose-dependently stimulated methyl-[3H]thymidine incorporation to MTC cells. These findings suggest a possible role of IGF-I as an autocrine growth factor for MTC cells. PMID- 2544166 TI - Protein phosphorylation and the two stages of pigment organelle dispersion in permeabilized xanthophores: organelle protein phosphorylation alone supports only the first stage. AB - We reported previously that, in cultured goldfish xanthophores, dispersion of aggregated carotenoid droplets (CDs) requires the specific phosphorylation of the CD protein p57 by a cAMP-dependent protein kinase and the presence of cytosol. We report here that, in permeabilized cells, the addition of the catalytic subunit of cAMP-dependent protein kinase and ATP phosphorylates p57 and converts the CDs from an immobile to a mobile state (first stage of CD dispersion). However, the CDs are restricted to the vicinity of the original site of the CD aggregate and do not actually disperse (second stage of CD dispersion) unless cytosol is also added. We propose that this process may be related to aspects of secretory processes. PMID- 2544163 TI - Photoaffinity labeling of hamster brown adipose tissue mitochondria by an [125I] coenzyme A derivative: differential interaction with the uncoupling protein and ADP/ATP carrier. AB - We have recently synthesized an azido [125I] CoA photolabel, N-(3-iodo-4 azidophenyl propionamide) cysteinyl-5-(2'thiopyridyl cysteine) CoA that specifically labeled the ADP/ATP carrier in beef heart mitochondria. In this study brown adipose tissue mitochondria were photolabeled with the azido [125I] ACT-CoA derivative with or without inhibitors. SDS gel electrophoresis and autoradiography of the separated proteins revealed exclusive photolabeling of two polypeptides corresponding to the ADP/ATP carrier and uncoupling protein. In the presence of carboxyatracytloside only the 32 kD UCP was labeled by [125I] ACT CoA, whereas preincubation with GDP resulted in exclusive photolabeling of the 30 kD ADP/ATP carrier. Palmitoyl CoA but not palmitic acid inhibited photolabeling of both polypeptides. PMID- 2544167 TI - Interactions between adenylate cyclase inhibitors and beta-adrenoceptors in isolated human fat cells. AB - Interactions between adenylate cyclase inhibitors and beta-adrenoceptors were investigated in isolated human fat cells. Phenylisopropyl adenosine, nicotinic acid and prostaglandine E2 induced a dose-dependent decrease in beta-adrenoceptor sensitivity; the concentration of isoprenaline causing half-maximum lipolytic effect increased 100-fold. The affinity constants for the high and low affinity beta-adrenoceptor states were increased 3000 and 700 times, respectively, but the total number of binding sites was unchanged. Pertussis toxin caused a dose dependent increase of beta-adrenoceptor sensitivity to isoprenaline. There was a 200-fold increase in isoprenaline sensitivity in the lipolysis experiments and corresponding increases in the receptor affinity in the binding experiments. It is concluded that the affinity of human fat cell beta-adrenoceptors is reduced by adenylate cyclase inhibitors. This seems to be mediated by the Gi-protein and represents a new potential mechanism by which lipolysis is regulated by inhibitors of adenylate cyclase in man. PMID- 2544168 TI - Introduction of cyclic AMP phosphodiesterase into rat submandibular acini prevents isoproterenol-stimulated cyclic AMP rise without affecting mucin secretion. AB - Cyclic AMP phosphodiesterase has been incorporated into isolated rat submandibular acini by hypotonic swelling. This resulted in complete inhibition of the cyclic AMP rise stimulated by isoproterenol (10 microM), but had no effect on the stimulation of mucin secretion. Acini swollen in the absence of cyclic AMP phosphodiesterase showed similar cyclic AMP and mucin secretion responses to those of unswollen acini. The dissociation between cyclic AMP rise and mucin secretion was not due to stimulation of different beta-receptor subtypes since both responses to isoproterenol were inhibited by the beta 1 antagonist atenolol, but not by the beta 2 antagonist, butoxamine. The results are the first to directly demonstrate that a maximally effective concentration of isoproterenol can increase mucin secretion in the absence of a detectable increase in cyclic AMP. PMID- 2544170 TI - The site of substrate and fructose 2,6-bisphosphate binding to rabbit liver fructose-1,6-bisphosphatase. AB - The binding site(s) in rabbit liver fructose-1,6-bisphosphatase for the active site binding ligand, fructose 6-phosphate, and the inhibitor, fructose 2,6 bisphosphate, have been investigated by using nuclear magnetic resonance spectroscopy. The distance from a nitroxide spin label to the bound ligands and the distance from the structural metal site to the bound ligands are about the same within experimental error. These data indicate that the two ligands probably bind at the active site in the rabbit liver enzyme. PMID- 2544171 TI - Inositol lipids and phosphatidic acid inhibit cell-free activation of neutrophil NADPH oxidase. AB - The effect of inositol lipids on the SDS-initiated cell-free activation of NADPH oxidase in membranes of human neutrophils was investigated. In a system consisting of low density membranes, cytosol and SDS, low doses of phosphatidylinositol, phosphatidylinositol mono- and biphosphates and phosphatidic acid interfered with activation of the oxidase. The inhibition was relieved by increasing concentrations of the cytosol. Conversely, preincubation of multilamellar phosphoinositide vesicles with cytosol reduced its ability to support activation of the oxidase. PMID- 2544172 TI - A possible role of superoxide anion radical in the process of blastocyst implantation in Mus musculus. AB - Superoxide anion radical and superoxide dismutase, the enzyme responsible for dismutating it, are both present in the ovary and uterus of Mus musculus during early pregnancy. The detectable, stable levels of superoxide radical and the constant high levels of superoxide dismutase in the ovary during early pregnancy suggests that these may be involved in the regulation of extended luteal steroidogenesis for the maintenance of pregnancy. An inverse correlation between the levels of superoxide anion radical and superoxide dismutase in the uterus is shown. The high levels of superoxide anion radical in the uterus on the early morning of Day 5 of pregnancy point towards a probable role for this radical in the act of implantation and in mediating the increased vascular permeability at the initiation of implantation. PMID- 2544169 TI - Ca2+ influx stimulated by vasopressin is mediated by phosphoinositide hydrolysis in rat smooth muscle cells. AB - The mechanism of Ca2+ influx stimulated by arginine vasopressin (AVP) was studied in cultured rat smooth muscle cells. AVP stimulated 45Ca2+ influx even in the presence of nifedipine, a Ca2+ antagonist that inhibits voltage-dependent Ca2+ channel. NaF, a GTP-binding protein activator, mimicked the AVP-stimulated 45Ca2+ influx. The 45Ca2+ influx stimulated by a combination of AVP and NaF was not additive. The affinity of AVP receptor was decreased by guanosine 5'-O-(3 thiotriphosphate). Pertussis toxin failed to affect the AVP-stimulated 45Ca2+ influx. AVP did not stimulate cAMP production, but increased inositol trisphosphate generation. Both AVP-stimulated 45Ca2+ influx and inositol trisphosphate generation were inhibited by neomycin, a phospholipase C inhibitor, in a dose-dependent manner, and the patterns of both inhibitions were similar. These results suggest that, in rat smooth muscle cells, AVP-stimulated Ca2+ influx is mediated exclusively through phosphoinositide hydrolysis. PMID- 2544173 TI - Coupling of a proton pump with superoxide radical-superoxide dismutase system in maturing mammalian spermatozoa and its association with sperm motility. AB - Mammalian spermatozoa possess a membrane-located 'proton pump' which appears to regulate the acquisition and maintenance of their motility. The 'proton pump' seems to be coupled with the superoxide radical-superoxide dismutase system which, when uncoupled, results in total loss of sperm motility. Addition of sulfhydryl-compounds restarts the proton pump and reactivates sperm motility. Cobaltous ions block the membrane sulfhydryl groups and it is proposed that the sulfhydryl groups may be involved in regulating proton secretion and sperm motility. PMID- 2544164 TI - Phorbol ester regulates the abundance of enkephalin precursor mRNA but not of amyloid beta-protein precursor mRNA in rat testicular peritubular cells. AB - Cultured peritubular cells prepared from the testes of 20-day-old rats contained both preproenkephalin (A) mRNA (1.5 kb) and amyloid beta-protein precursor mRNA (3.6 and 2.8 kb). The phorbol ester TPA and forskolin (an adenylate cyclase activator) increased the preproenkephalin mRNA abundance to 9.0 and 5.8 times the control, respectively. TPA alone had no effect on the intracellular cAMP level. A combination of TPA and forskolin elicited a synergistic increase in the ppEnk mRNA abundance over 30-fold. Dexamethasone potentiated the effect of forskolin but not of TPA. These results suggest that TPA regulates the preproenkephalin mRNA abundance through a cAMP-independent pathway. In contrast, TPA, forskolin, and dexamethasone showed little or no effect on the abundance of amyloid beta protein precursor mRNA. PMID- 2544174 TI - High and low affinity binding sites for endothelin on cultured rat glomerular mesangial cells. AB - Endothelin contracts glomerular mesangial cells, thereby influencing glomerular size and filtration rate. Here, we demonstrate the presence of two ET-specific binding sites on cultured rat mesangial cells with Kds of 0.76 and 44.70 nM, and maximal binding capacity (Bmax) values of 6.78 x 10(2) and 27.60 x 10(2) binding sites/cell, respectively. Binding of [125I]-ET was maximal at 120 min at 4 degrees C, stable for the subsequent 60 min, and selective. No competition for binding was observed with greater than 1000-fold concentrations of atrial natriuretic peptide, angiotensin II, arginine vasopressin, nicardipine, or nifedipine. The presence of specific receptors for ET on glomerular mesangial cells suggests a major role for this peptide in the regulation of glomerular filtration rate. PMID- 2544177 TI - Mechanism of the reversal reaction of platinated DNA with thiourea studied by platinated 5'-GMP. AB - The reversal reactions of cis-[Pt(NH3)2(5'-GMP)2] 2-(1) and trans-[Pt(NH3)2(5' GMP)2] 2-(2) with thiourea were examined by reversed phase HPLC and monothioureido intermediate cis-[Pt(NH3)2(5'-GMP) (tu)] (4) was detected. This result suggested that Pt-[5'-GMP-N(7)] bond was more labile than Pt-NH3 bond and the release of ammonia from cis-Pt(II)-DNA base complexes is a result of trans labilizing effect of sulfur containing molecule displaced with DNA base. PMID- 2544175 TI - Kinetics of ribosomal protein S6 phosphorylation by HepG-2 cells in response to insulin. AB - The effect of insulin on the phosphorylation of ribosomal protein S6 was studied in a human liver cell line (HepG-2), using [32P] inorganic phosphate. Increased rate of protein S6 phosphorylation was detected 8 min following the addition of insulin to serum starved cells. Maximum enhancement of phosphorylation was observed at 80 nM insulin. Minimum level of insulin required to produce measurable increase of S6 phosphorylation was 20 nM. Radioactivity of protein S6 increased most in the native subunit and polysome fractions. Significant increase in radioactivity of this protein was not observed in the monosome fraction during the first 30 min of insulin stimulation. Increase in the specific radioactivity of native 40S subunit was higher than that of polysomes. These results suggest that phosphorylation takes place in the subunit compartment and moves preferentially into the polysomes. PMID- 2544179 TI - Expression of genes of ferritin subunits in human hepatoma cell lines. AB - The expression of ferritin genes has been studied in two well differentiated hepatoma cell lines, HepG2 and Hep3B, and in an undifferentiated cell line, HepPLC/PRF/5. The steady-state level of the H and L ferritin subunit mRNAs is different in the three cell lines, being most abundant in the HepG2 and Hep3B cells. The efficiency of promotion of transcription of the H and L gene promoters is not correlated with the levels of the two transcripts; moreover the half-life of the H and L mRNAs varies among the hepatoma cells. The different level of ferritin mRNAs in differentiated and undifferentiated cells appears to be mainly due to a different stability of the transcripts. PMID- 2544176 TI - Endothelin inhibits presynaptic adrenergic neurotransmission in rat mesenteric artery. AB - The effect of endothelin(ET) on adrenergic neurotransmission was examined in isolated perfused rat mesenteric arteries. Porcine ET(10(-12) to 10(-10)M) attenuated the pressor response to sympathetic nerve stimulation (NS). It also stimulated the release of prostaglandin E2 (PGE2), but its inhibition of the pressor response to NS was not affected by indomethacin treatment. ET also caused dose-dependent inhibition of [3H]norepinephrine release during NS. Higher doses of ET rather enhanced the pressor response to NS. These results suggest that ET inhibits presynaptic adrenergic neurotransmission without mediation of PGE2, while it potentiates the responsiveness of the postsynaptic alpha-adrenergic receptor. Thus ET appears to act directly on the neuroeffector junction as well as on the peripheral vasculature. PMID- 2544178 TI - Phosphatase inhibition with okadaic acid does not alter the relationship between force and myosin light chain phosphorylation in permeabilized smooth muscle. AB - The phosphatase inhibitor, okadaic acid, has been used to test the hypothesis that myosin light chain phosphatase activity plays a central role in latchbridge formation in smooth muscle. In the permeabilized rabbit portal vein there is a non-linear relationship between myosin light chain phosphorylation and force production such that maximum force output occurs with about 50% phosphorylation. Treatment of the muscle with okadaic acid does not change this relationship even though there is a profound inhibition of phosphatase activity. The data suggest that dephosphorylation of the myosin light chain while the myosin is in the force producing state does not account for the high force output with low levels of light chain phosphorylation in smooth muscle. PMID- 2544183 TI - Topoisomerase II-mediated DNA cleavage activity induced by ellipticines on the human tumor cell line N417. AB - Ellipticine derivatives have been shown to induce DNA strand breaks by trapping DNA-topoisomerase II (Topo II) in an intermediary covalent complex between Topo II and DNA which could be related to their cytotoxic effects. We report here that Celiptium and Detalliptinium, two ellipticine derivatives clinically used for their antitumoral properties against breast cancer, exhibit the highest in vitro activity on Topo II DNA cleavage reaction and decatenation among a series of 14 ellipticine derivatives. The in vitro cleavage site specificity in pBR 322 plasmid DNA and in a human c-myc gene inserted in a lambda phage DNA is identical for both ellipticines, but different from m-AMSA, another Topo II related antitumoral agent. Recently, it has been shown that the ellipticine derivative Celiptium presents a strong cytotoxic activity in vitro on different human tumors including small cell lung carcinoma (SCLC). However, the studies that involved Topo II as a target for ellipticine derivatives have been performed only by using animal tumor cell lines. Therefore we have studied the in vivo DNA cleavage activity of Celiptium and Detalliptinium on a human SCLC cell line, NCI N417, comparatively to that obtained with m-AMSA. The respective IC50 on cell growth are 9, 8 and 1 microM for Celiptium, Detalliptinium and m-AMSA, respectively. Using the alkaline elution technique, we have observed that Celiptium and Detalliptinium exhibit a weak cleavage activity on genomic DNA from whole cells. The ellipticines are about 50 times less potent than m-AMSA in inducing DNA strand breaks. Analysis of in vivo c-myc gene cleavage by Southern blot hybridization also demonstrates a lack of activity of the ellipticine derivatives as no gene cleavage could be detected up to 50 microM of the drug. With m-AMSA, c myc gene cleavage is detected at a concentration of 0.2 microM, which indicates that this methodology is less sensitive in detecting DNA strand breaks than is the alkaline elution. Further studies of the drug effect on isolated nuclei by alkaline elution also show that the DNA cleavage activity of Celiptium and Detalliptinium is increased when compared to whole cells. Our data indicate that these two drugs have a weaker cytotoxic effect than m-AMSA on NCI N417 cell line, due to a limited access to the cell nucleus rather than to a lack of activity on Topo II as assessed by in vitro and isolated nuclei experiments. PMID- 2544180 TI - Differential sensitivity of phosphoinositide and cyclic GMP responses to short term regulation by a muscarinic agonist in mouse neuroblastoma cells. Correlation with down-regulation of cell surface receptors. AB - Short-term agonist-induced loss of cell surface muscarinic receptors and desensitization of receptor-mediated cyclic GMP (cGMP) formation and phosphoinositide hydrolysis were examined in mouse neuroblastoma cells (clone N1E 115) in suspension. This treatment resulted in a time-dependent reduction of approximately 40% of the specific binding of the hydrophilic antagonist [3H]N methyl-scopolamine [( 3 H]NMS) with a T 1/2 of down-regulation of 4.83 min. Scatchard analysis revealed that brief exposure to the agonist resulted in a significant reduction in the Bmax with no change in the Kd. Agonist-induced cGMP formation decreased in a similar time-dependent manner with an average T 1/2 of 4.79 min. However, desensitization of muscarinic receptor-stimulated accumulation of inositol phosphates demonstrated a much slower time-course and was accompanied by a reduction in the maximal response with no change in the EC50. In addition, there was rapid partial recovery of cell surface receptors and desensitized cGMP response, with no apparent resensitization of phosphoinositide hydrolysis. Thus, there was a differential rate of short-term desensitization and resensitization of these two muscarinic receptor-mediated responses. Moreover, desensitization of cGMP formation, but not phosphoinositide hydrolysis, closely paralleled loss of cell surface muscarinic receptors. PMID- 2544184 TI - Plasma DNA in systemic lupus erythematosus. Characterization of cloned base sequences. AB - Little is known about plasma DNA in patients with systemic lupus erythematosus (SLE). Previous studies have suggested that it may, in fact, not be derived from the random nucleolysis that might be expected to accompany cell death, which might have been the simplest explanation of its origin. To extend the results of other studies, plasma DNA, obtained from a subgroup of SLE patients who had large amounts of immunoprecipitable plasma DNA, was cloned into a plasmid vector, and the nucleotide base sequences were studied by nucleic acid hybridization. One possible explanation for the anomalous renaturation kinetics of plasma DNA in SLE -namely, that it contained non-human genomic base sequences--was rendered untenable by the fact that 51 consecutive DNA clones from 2 SLE patients were each shown to contain human genomic DNA sequences. Plasma DNA from SLE patients also differed from human genomic control DNA in that those sequences derived from the highly repetitive fraction of human DNA were specifically enriched in one such sequence, Alu, which constituted 55% of the repeat sequences in the plasma DNA clones as compared with 13% in control DNA clones. There was a decrease in the frequency of non-Alu repeat sequences (9% for plasma DNA compared with 23% in control DNA). These differences were statistically significant, and they indicate that SLE plasma DNA contains a nonrandom selection of human genomic base sequences. Several explanations for these findings are considered. PMID- 2544181 TI - Inhibitory effect of 8-bromo cyclic GMP on an extracellular Ca2+-dependent arachidonic acid liberation in collagen-stimulated rabbit platelets. AB - The inhibitory effect of cyclic GMP on collagen-induced platelet activation was studied using 8-bromo cyclic GMP (8brcGMP) in washed rabbit platelets. Addition of collagen (1 micrograms/ml) to platelet suspension caused shape change and aggregation associated with thromboxane (TX) A2 formation. 8brcGMP (10-1000 microM) inhibited collagen-induced platelet aggregation and TXA2 formation in a concentration-dependent manner. 8brcGMP did not affect platelet cyclooxygenase pathways, but markedly inhibited collagen-induced arachidonic acid (AA) liberation from membrane phospholipids in [3H]AA-prelabeled platelets, indicating that the inhibitory effect of 8brcGMP on collagen-induced aggregation is due to an inhibition of AA liberation. In [32P]orthophosphate-labeled platelets, collagen stimulated phosphorylation of a 20,000 dalton (20-kD) and 40-kD proteins. 8BrcGMP stimulated phosphorylation of a specific protein having molecular weight of 46-kD and inhibited collagen-induced both 20- and 40-kD protein phosphorylation. Collagen could stimulate the AA liberation without activation of phospholipase C or Na+-H+ exchange, but could not in the absence of extracellular Ca2+. These findings suggest that cyclic GMP inhibits collagen induced AA liberation which is mediated by an extracellular Ca2+-dependent phospholipase A2. However, cyclic GMP seems to inhibit the Ca2+-activated phospholipase A2 indirectly, since 8brcGMP had no effect on Ca2+ ionophore A23187 induced platelet aggregation or AA liberation. It is therefore suggested that cyclic GMP may regulate collagen-induced increase in an availability of extracellular Ca2+ which is responsible for phospholipase A2 activation in rabbit platelets. PMID- 2544189 TI - Age- and sex-related differences in antigen-induced arthritis in C57Bl/10 mice. AB - The influence of both age and sex on antigen-induced arthritis in C57Bl/10 mice was studied. Methylated bovine serum albumin was used to induce arthritis in young adult (3 months) and old (18 months) male and female mice. Arthritis became chronic and led to severe joint damage more often in 18-month-old female mice, compared with both young adult female mice and with male mice of both ages. T cell immunity and levels of antibodies against methylated bovine serum albumin were comparable in all groups. Antigen retention in the joint was greater in old compared with young adult mice of both sexes, and could therefore not entirely explain the increased prevalence of persistent arthritis in old females. Our data suggest that female hormones, in combination with age, are important factors in the chronicity and destructive character of this type of arthritis. PMID- 2544182 TI - The curvilinear Scatchard plot. Experimental artifact or receptor heterogeneity? PMID- 2544188 TI - Acute peritendinitis calcarea. PMID- 2544185 TI - Clinical heterogeneity and treatment response in inclusion body myositis. AB - Inclusion body myositis has been described as an inflammatory myopathy with distinctive clinical and pathologic features that is refractory to treatment. Ten cases of inclusion body myositis, as defined by histopathologic findings, were reviewed to determine whether the clinical characteristics are different in patients whose disease has been defined by light and electron microscopic studies compared with those whose disease has been defined by light microscopic studies alone. The clinical characteristics of both groups of patients were similar, and 2 patients have had excellent responses to treatment. Although inclusion body myositis represents a histologic subset of polymyositis, from a clinical perspective, it must be considered a nonspecific designation. Despite a generally poor prognosis, therapeutic intervention is still warranted. PMID- 2544186 TI - Hydroxyapatite pseudopodagra. A syndrome of young women. AB - Six women with acute calcific periarthritis of the first metatarsophalangeal joint (hydroxyapatite pseudopodagra) are described, and 10 previously reported cases (8 female patients and 2 male patients) are reviewed. The onset was characterized by acute pain, swelling, erythema, tenderness, and limitation of movement of the first metatarsophalangeal joint, symptoms indistinguishable from those of gouty podagra, which is associated with transient amorphous calcific deposits in the vicinity of the joint. Based on these results it appears that hydroxyapatite pseudopodagra is a disorder that predominantly affects premenopausal women. PMID- 2544190 TI - Normal serologic response to Epstein-Barr virus in patients with Sjogren's syndrome. PMID- 2544187 TI - Prophylactic treatment of canine osteoarthritis with glycosaminoglycan polysulfuric acid ester. AB - The prophylactic effect of glycosaminoglycan polysulfuric acid ester (GAGPS) on cartilage lesions was studied using the Pond-Nuki model of canine osteoarthritis. Starting 2 days after anterior cruciate transection, GAGPS or saline was administered intraarticularly twice weekly for 4 weeks. After 4 weeks, gross and histologic medial femoral condylar lesions had developed to a lesser degree in GAGPS-treated dogs than in saline-treated dogs. The uronic acid and hydroxyproline levels in cartilage were significantly higher in the GAGPS-treated dogs than in the saline-treated dogs. Levels of active and latent collagenase in the cartilage of GAGPS-treated dogs were lower than in the cartilage of saline treated dogs. With GAGPS treatment, swelling of the cartilage, an indicator of collagen network integrity, remained near control levels. Although increased synthesis of proteoglycan and collagen may account for some of these results, we propose that one mechanism of action of GAGPS is its ability to decrease collagen degradation, either by decreasing the synthesis of collagenase or by directly inhibiting the production of collagenase in cartilage. PMID- 2544193 TI - X-ray absorption spectroscopic investigations of cytochrome c oxidase structure and function. AB - Although the low sensitivity of the XAS technique imposes difficulties upon the study of an enzyme that is often heterogeneous, significant progress has been made in elucidation of the structures of the functional metal sites of cytochrome c oxidase. Figure 8 summarizes interpretations of the resting-state enzyme based on the XAS results obtained over the past decade by the two main groups involved. Aside from several persisting minor differences in distances and precise ligand compositions of the metal sites, the biggest difference between the two interpretations involves the binuclear O2 interaction site, especially the nature of the bridging ligand and its precise attachments to Fea3 and CuB. However, the structural models presented by the two groups have been converging recently, and there is hope that the next few years will see a resolution of the remaining differences. Other XAS approaches (e.g. studies on oriented multilayers) and other techniques will doubtless contribute to this resolution. PMID- 2544192 TI - [Detection of papillomaviruses by in situ hybridization with sulfonated probes]. AB - A technique of detection by in situ hybridization of human papillomavirus in sections of condylomatous lesions is described. The probes are labeled and modified by sulfonation and the hybrids are revealed by immunohistochemistry, using alkaline phosphatase. PMID- 2544194 TI - Peroneal nerve dysfunction as a complication of lateral meniscus repair: a case report and anatomic dissection. AB - A case report is presented in which a complete peroneal nerve palsy complicated lateral meniscus repair. The meniscal repair was performed using the combined method of arthroscopically placed inside-out needles and a posterior incision to view their exit. A cadaver limb was then used to examine the anatomy of lateral meniscus repair. Needles used for meniscus repair were passed into the posterior horn of the lateral meniscus using the inside-out and the outside-in methods. Computerized axial tomography (CAT) was then used to establish the proximity of the needles to the peroneal nerve in the posterolateral corner. The inside-out needles were dangerously close to the peroneal nerve. The outside-in needles had a larger margin of safety. The posterolateral corner was then dissected to verify and photograph these relationships. PMID- 2544195 TI - Structural and functional aspects of platelet-derived growth factor and its role in the pathogenesis of glioblastoma. AB - The platelet-derived growth factor (PDGF) family consists of three different dimeric forms, AA, BB, and AB, of the two constituent polypeptide chains, A and B. These interact with two different cell surface receptors that, in part, mediate different cellular functions. The various forms of PDGF, as well as the receptors, are expressed at high frequency in glioblastoma multiforme, and it has been suggested that the growth of this tumor might be affected by autocrine loops involving PDGF and its receptors. The present paper focuses on recent discoveries regarding the family of PDGF ligands and receptors, as well as reviews results concerning PDGF-dependent autocrine growth in experimental and spontaneous glioblastoma. PMID- 2544191 TI - [Composite carcinoma of the prostate combining a small cell carcinoma and an adenocarcinoma. Apropos of a case]. AB - A case of combined adenocarcinoma and small cell carcinoma of the prostate is described in a 58-year-old-man. Prostatic acid phosphatases and neuron specific enolase were found elevated in the serum. At autopsy the lung was free of tumor. The liver was replaced by numerous metastatic nodules and a voluminous mesenteric metastasis extended into the wall of the vessels (aorta and vena cava). Microscopic examination showed a small cell carcinoma component of the oat cell type and an adenocarcinoma component constituting 10% of the total tumor volume. By immunostaining, the small cell carcinoma component is neuron specific enolase+ and prostatic specific antigen-. The adenocarcinoma component is neuron specific enolase- and prostatic specific antigen+. PMID- 2544196 TI - Amino acid sequence of cytochrome c from Aspergillus niger. AB - Cytochrome c from Aspergillus niger consists of two forms, a major one (80%) with 111 amino acid residues and a minor one (20%) with 108 residues, missing the three N-terminal residues of the major one. The primary sequence of A. niger cytochrome c was determined by standard spinning-cup Edman degradation of purified peptides and of pairs of peptides, from which the desired sequence was readily deduced by subtraction of common sequencies. Except for the extension and some variability at the N-terminal sequence, the A. niger protein conforms well with other cytochrome c structures. PMID- 2544199 TI - Functional maturation of the GABAergic inhibition on dopamine-mediated behaviours during the neonatal period in the mouse. AB - Previous works have indicated that systemic injection of GABA-agonists depress motoric behaviours in neonatal murids, suggesting an early maturation of GABAergic inhibitory processes. In this paper, the inhibitory effects of muscimol, a postsynaptic GABAA-agonist, on D-amphetamine-induced enhancement of locomotion, wall-climbing and head-raising were examined in neonatal 5-, 8- and 11-day-old mouse pups, using a direct observational procedure. The results show that muscimol can selectively attenuate high levels of locomotion, wall-climbing and head-raising produced by the indirect dopamine agonist in 8- as well as 11 day-old pups. However, while muscimol is able to moderate amphetamine-induced wall-climbing and head-rising in 5-day-old pups, no GABAergic inhibition was seen for locomotion at this age. Licking episodes elicited by amphetamine in 11-day old pups can be magnified by muscimol if the dosage of the former is relatively too potent. It is suggested that the GABAergic inhibitory processes on dopaminergic functioning have reached good levels of functional maturation in the neonatal murid. PMID- 2544198 TI - An investigation of some temporal aspects of olfactory coding with the model of multi-site electrical stimulation of the olfactory bulb in the rat. AB - Electrical stimulation of the olfactory bulb was used to investigate some temporal aspects of olfactory coding, with reference to respiration. Food deprived rats implanted with permanent electrodes were trained to use bulbar multi-site stimulation patterns as discriminative stimuli for predicting the nature of an incoming reinforcement. Electrical pulse trains (100 Hz) were periodically delivered in phase with precisely defined moments of the respiratory cycle (during inspiration or expiration). Temporal aspects of olfactory coding were first considered through the measurement of the minimum duration of a stimulus necessary to identify this stimulus. The results showed that a bulbar stimulation lasting for 30 ms (3 pulses), and delivered during inspiration, was clearly identified by the rats. Stimulus identification induced a discriminative respiratory response which could manifest itself as early as the first cycle concomitant with the beginning of stimulation. It was then shown that a bulbar electrical stimulation pattern was identified with the same latency whether it occurred during expiration or during inspiration. Moreover, the perceptive events induced in those two conditions of stimulation were not different enough to be discriminated by the animals. The findings are discussed within the framework of olfactory information processing. PMID- 2544197 TI - Effects of the opiate antagonist naloxone upon hypothalamically elicited affective defense behavior in the cat. AB - The opiate antagonist naloxone hydrochloride was employed in order to determine whether endogenous opioids play a role in the control of affective defense behavior elicited from the medial hypothalamus in the cat. The effects of naloxone upon quiet biting attack behavior elicited from the lateral hypothalamus were also assessed. A comparison of the differences in response latencies or thresholds before and after naloxone (i.p.) administration was made. Naloxone (1, 4 and 10 mg/kg) was found to significantly facilitate affective defense behavior in a dose- and time-dependent manner. The duration of facilitation ranged from 30 min after a 1 mg/kg injection to 180 min after a 10 mg/kg injection. The data also suggest that the effects of naloxone upon affective defense behavior are opposite to those seen with quiet biting attack. In two animals, quiet biting attack behavior was suppressed for 30 min following a 10 mg/kg injection of naloxone. Naloxone was also administered to cats in which hypothalamic stimulation elicited predatory responses coupled with components of affective defense behavior. In these cases, naloxone was ineffective in altering latencies for this 'mixed' response. These findings suggest that the opiate peptide system selectively inhibits affective defense behavior elicited from the medial hypothalamus of the cat. PMID- 2544200 TI - Footshock-induced sensitization of electrically elicited startle reflexes. AB - The acoustic startle reflex can be facilitated by the presentation of a train of footshocks presented in rapid succession (footshock sensitization). Acoustic startle is a short-latency reflex mediated by a neural circuit consisting of the ventral cochlear nucleus (VCN), ventral nucleus of the lateral lemniscus (VLL), the nucleus reticularis pontis caudalis (RPC), and the spinal cord. The present study sought to determine the point along this pathway where footshocks might ultimately alter neural transmission to affect startle response. Rats were implanted bilaterally with stimulating electrodes in either the VCN, VLL, or RPC. Startle could be elicited acoustically with a noise burst or electrically with a single-pulse stimulus to either the VCN, VLL, or RPC before and after a train of ten 0.6-mA, 500-ms shocks presented at a rate of 1 shocks/s. Startle elicited acoustically or electrically in the VCN or VLL was significantly elevated following shocks. In contrast, startle elicited from the RPC showed no sensitization, even though startle elicited acoustically from the same animals during the same test session was facilitated. These data suggest that footshock sensitization ultimately alters transmission in the startle circuit at the RPC. PMID- 2544201 TI - Sensitization of the startle reflex by footshock: blockade by lesions of the central nucleus of the amygdala or its efferent pathway to the brainstem. AB - Bilateral electrolytic lesions of the central, but not the lateral, nucleus of the amgydala blocked shock sensitization of startle (the increase in startle produced by presentation of ten 0.6-mA footshocks in rapid succession). Lesions of the central nucleus also decreased reactivity to shock (jumping and flinching) during shock presentation. However, this decrease in reactivity cannot account for the blockade of shock sensitization, because when a higher shock intensity (1.0 mA) was used, producing equivalent reactivity to that of controls at 0.6 mA, central nucleus lesions still blocked shock sensitization. Moreover, lesions of the caudal part of the ventral amygdalofugal pathway, which carries central nucleus efferents to the startle reflex pathway, also blocked shock sensitization. It is hypothesized that shock activates the central nucleus of the amygdala, which increases startle through modulation of the startle pathway. Activation of the amygdala by shock may be the unconditioned response relevant for fear conditioning. PMID- 2544202 TI - Reexamination of the gill withdrawal reflex of Aplysia californica Cooper (Gastropoda; Opisthobranchia). AB - The gill withdrawal reflex (GWR), an important model system for neural mechanisms of learning, varies in form and amplitude within as well as between preparations and is therefore a heterogeneous collection of action patterns, not a reflex. At least 4 action patterns occur in response to mechanical stimulation of the siphon. It is often impossible to categorize a particular movement unambiguously. All may occur spontaneously. Gill movements may be described as combinations of 10 actions; 4 involving vein movements are described here. All actions and action patterns can occur in preparations lacking the central nervous system. Some vein movements may generate considerable force without markedly altering gill area. It is suggested that this explains why some early studies failed to identify the important role of the peripheral nervous system in the GWR. Studies based on the assumption that the GWR involves a single type of movement controlled by cells of the parietovisceral ganglion require reevaluation. PMID- 2544203 TI - Conditioned taste aversions are not readily disrupted by external excitation. AB - Thirsty male rats were given saccharin water followed by delayed illness. During the delay, some of the rats were exposed to events designed to stimulate their external systems (i.e., the system that processes external events such as auditory and tactile stimulation). Access to females, mild footshocks, and pain from hypertonic saline injections did not interfere with either the acquisition or extinction of a taste aversion. In fact, when administered intraperitoneally, the hypertonic saline slightly increased the strength of the aversion. Exposure to heat, which changed both skin temperature and core temperature, slightly attenuated the formation of the aversion. Overall, these results emphasize the independence of the internal system (i.e., the system that deals with internal events such as taste, illness, and core temperature) and the external system. Furthermore, the associating of events related to the internal system is not readily interfered with by events related to the external system. PMID- 2544204 TI - Lateral hypothalamic stimulation can augment or attenuate nucleus gigantocellularis escape: evidence for appetite-associated aversion amelioration. AB - Carr and Coons (1982a, 1982b) found that lateral hypothalamic (LH) stimulation ameliorates the aversiveness of stimulation of pain-implicated nucleus gigantocellularis (NGC), but this finding disagrees with other findings. To resolve this disagreement, we tested whether amelioration is differentially associated with the ability of LH stimulation to support self-stimulation (SS), to support responding to escape LH stimulation (LH escape), or to elicit stimulation-bound feeding (SBF). LH stimulation not yielding SBF always increased responding to escape from NGC stimulation (NGC escape) and was reward-escape in nature in supporting LH escape as well as SS. By contrast, LH stimulation yielding SBF always reduced NGC escape and was purely rewarding in that it only supported SS and never LH escape. In an additional experiment, the anxiolytic diazepam augmented the ability of LH stimulation yielding SBF to reduce NGC escape. PMID- 2544205 TI - Role of the pituitary-adrenal hormones in the acquisition of schedule-induced polydipsia. AB - Adrenalectomized female rats failed to develop schedule-induced polydipsia (SIP). Dexamethasone (DEX) injections failed to reinstate SIP in adrenalectomized rats. They did not prevent intact rats from acquiring SIP but interfered with subsequent expression of this behavior. In contrast, corticosterone, the rats' normally occurring glucocorticoid, fully restored the acquisition and subsequent expression of SIP in adrenalectomized rats. This strongly suggests that corticosterone plays an essential role in the normal acquisition and development of this behavior. Data are interpreted in the context of current information concerning adrenal hormone receptors. It is hypothesized SIP acquisition is at least partly regulated by the Type I (mineralocorticoid) receptor. PMID- 2544206 TI - Conditional immunomodulation following training with cyclophosphamide. AB - In 5 experiments, paired-group rats received a conditional stimulus (CS) paired with the immunosuppressive drug cyclophosphamide (CY). In Experiments 1-3, the CS was saccharin (SAC). Consistent with previous reports, these rats acquired a SAC aversion. However, there was no evidence of conditional immunosuppression. Rather, when reexposed to SAC in conjunction with an antigenic challenge, paired group rats evidenced hemagglutination antibody titers similar to those seen in rats that never received the immunosuppressant. That is, the usual effect of CY in compromising immunological functioning was attenuated or eliminated by the CY paired flavor. The findings of Experiments 1-3 were confirmed in Experiments 4-5, which used nongustatory CSs. Both audiovisual (noise and flashing-light) and pharmacological (pentobarbital) cues were also effective signals for CY injection. Following pairing with CY, these cues protected animals from the immunosuppressive effects of the drug. PMID- 2544207 TI - Modulation of appetitively and aversively motivated behavior by the kappa opioid antagonist MR2266. AB - MR2266 (MR), an opioid antagonist that binds to kappa and mu receptors, was examined for its ability to influence the aversively motivated behaviors conditioned by electric shock and the drinking induced by water deprivation or the availability of a palatable saccharin/glucose solution. The intraperitoneal (ip) and intracerebroventricular (icv) administration routes were contrasted. After both ip and icv administration, MR was able to reverse conditional analgesia as measured by the formalin test. MR enhanced the Pavlovian conditional freezing response when administered icv prior to shock exposure but reduced freezing if given ip prior to shock. A related benzomorphan-derived opioid antagonist, MR1452, also reduced freezing when given ip prior to shock. MR2266 was a potent antidipsogenic agent when administered ip but had no such effect when administered icv. It is concluded that separable opioid systems are involved in the modulation of appetitively and aversively motivated behaviors. PMID- 2544209 TI - Effects of malotilate treatment on alcoholic liver disease. AB - Malotilate, a new hepatotrophic drug, improves serum transaminase levels and the markers of protein metabolism in the liver in chronic liver diseases. However, the effects of malotilate on alcoholic liver disease are not well known. In the present study, the effects of this drug on the recovery process of alcoholic liver disease after abstinence were analyzed. Many hepatic test values were significantly improved after abstinence from alcohol in both the malotilate treated and nontreated control groups. However, the Normotest values improved significantly only in the malotilate group, and not in the control group. The improvement rates for choline esterase activity were significantly greater in the malotilate group than in the control group. Serum albumin levels significantly increased in the malotilate group but not in the control group. Changes in the serum markers of hepatic fibrogenesis were not different between the 2 groups. These results indicate that malotilate accelerates the recovery of impaired protein metabolism in alcoholic liver disease and that this drug may be useful for the treatment of alcoholic liver diseases. PMID- 2544208 TI - Retention of maze performance inversely correlates with N-methyl-D-aspartate receptor number in hippocampus and frontal neocortex in the rat. AB - The N-methyl-D-aspartate (NMDA) receptor may play a critical role in learning and memory. In the present study, a significant correlation was found between the number of NMDA-displaceable, Na+-independent L-[3H]glutamate binding sites in the hippocampus and neocortex of young rats and the mean number of errors during retention, but not acquisition, in a 14-unit T-maze. PMID- 2544211 TI - Nuclear DNA distribution in neuroendocrine gastroenteropancreatic tumors before and during treatment. AB - The nuclear DNA contents of tumor cells in 73 patients with endocrine gastrointestinal tumors, 19 patients with endocrine pancreatic tumors (EPT) and 54 patients with malignant carcinoid tumors were determined before and after treatment. The DNA profiles were divided into diploid and aneuploid. In untreated patients, 9 out of 10 (90%) primary EPT and all 9 primary malignant carcinoid tumors (100%) were diploid. Tumor cell imprints from liver metastases of patients with untreated EPT showed aneuploidy in 5 of 11 cases, but only in 7 out of 46 DNA records from patients with untreated carcinoid liver metastases. DNA alteration from diploid to aneuploid profiles occurred in 2 patients with endocrine pancreatic tumors who had received chemotherapy. A change from diploid to aneuploid records was also seen in 7/23 (30%) carcinoid tumors after treatment. The DNA patterns before and after treatment did not show any correlation with survival or treatment response. PMID- 2544210 TI - Ro15-4513 enhances and attenuates motor stimulant effects of ethanol in rats. AB - The actions of the imidazobenzodiazepine, Ro15-4513 (2.5 mg/kg IP), ethanol (0.50 g/kg, 0.75 g/kg, 1.25 g/kg), and Ro15-4513 in combination with ethanol were assessed using low activity male Charles River (CD) rats. Horizontal (ambulatory) and nonhorizontal (rearing, grooming, etc.) activity were measured in the open field with a Digiscan activity system. When Ro15-4513 was given alone, animals responded in a manner similar to that of the control animals. Ethanol alone, however, resulted in a significant enhancement of activity, with the effect being most pronounced at 0.50 g/kg for horizontal activity and 1.25 g/kg for nonhorizontal activity. A potentiation of the ethanol-induced stimulant effect was observed following pretreatment with Ro15-4513 on the horizontal activity measure for the 1.25 g/kg ethanol group. An attenuation of activity was observed for this group on the nonhorizontal activity measure. Ro15-4513, however, did not affect the stimulation produced by the other doses of ethanol on the nonhorizontal measure. The results suggest that possibility of distinct neurochemical mechanisms in the mediation of horizontal and nonhorizontal activity and that, perhaps, GABA-benzodiazepine mechanisms may play different roles in mediating their ETOH-induced stimulant effect. PMID- 2544212 TI - Nitrosoureas. Modes of action and perspectives in the use of hormone receptor affinity carrier molecules. AB - Mechanisms of DNA adduct formation by antineoplastic 2-chloroethyl-N-nitrosoureas (CNUs) and of DNA damage induced by these compounds are discussed. CNUs are alkylating agents that form DNA-DNA cross-links as well as 2-chloroethylated and 2-hydroxyethylated adducts, the N-7-position of guanine being the predominantly alkylated site. A close correlation exists between the potential of a given compound to induce DNA-DNA cross-links and its antineoplastic effectiveness. However, levels of DNA-DNA cross-linking in bone marrow and extent of myelosuppression as measured in rodents are also closely correlated. The design of new cross-linking analogues capable of directing the antineoplastically relevant activity predominantly to the target tumour appears therefore to be of great promise. Cross-linking agents have been attached to a variety of steroid hormone carrier molecules and the conjugates have been tested in structure activity studies using hormone-receptor containing animal tumours. These studies have revealed that some hormone-linked antineoplastic agents are highly effective in receptor positive experimental tumours and are superior to mixtures of unlinked alkylating agents with hormones. Indications for a relative enrichment of DNA damaging effects in the tumour tissue and for reduced myelotoxicity have been obtained with specific hormone conjugates. PMID- 2544213 TI - [Evaluation of a rapid diagnosis of congenital lactic acidemia by transported peripheral blood samples]. AB - The usefulness of a rapid diagnosis of congenital lactic acidemia was investigated using peripheral blood samples from 40 patients with lactic acidemia which had been transported from the hospitals in every part of Japan. Platelets and monocytes were separated, and the rates of decarboxylation of pyruvate and activities of enzymes involved in pyruvate metabolism were measured. The activity of phosphoenolpyruvate carboxykinase in monocytes was relatively stable. However, [1-14C] pyruvate and [3-14C] pyruvate decarboxylation rates and cytochrome c oxidase activity in platelets and pyruvate carboxylase activity in monocytes were unstable and decreased during the transportation of blood samples. Therefore, in order to diagnose the enzyme defects, it was necessary to compare the values for patients with those for control subjects who were simultaneously examined. Using this method, a patient with pyruvate dehydrogenase complex deficiency was found in the 40 patients with congenital lactic acidemia. PMID- 2544214 TI - Towards standard sera for the determination of anti-neutrophil cytoplasmic (ANCA) and anti-myeloperoxidase (aMPO) antibodies. PMID- 2544215 TI - Interaction of HIV and EBV at lymphoid tissue level: immunohistochemistry and in situ hybridization. AB - The presence of Epstein-Barr virus antigens and genome was studied in lymph nodes from HIV + patients affected by PGL. Cryostat sections from 50 lymph nodes of HIV + patients were immunostained with EBV-VCA (viral capsid antigen), EBV-EA (early antigen) and p24 HIV major core protein monoclonal antibodies. In situ hybridization was performed using a biotin conjugated EBV DNA probe; the reaction product was demonstrated by immunohistochemical method. As positive controls, EBV producer B95-8 and HIV infected H9 cell lines were used. The majority of patients had circulating EBV antibodies mainly directed against the viral capsid antigen and only in few cases against early antigens. Positivity for HIV p24 protein was detected in 43 out of 50 lymph nodes within the germinal centers with a reticular pattern. Only 2 out of 50 lymph nodes presented very few positive cells for EBV antigens and none expressed detectable EBV genome. Our results suggest that EBV cellular expression does not correlate with serum positivity; furthermore the absence of EBV antigens and genome at tissue level might indicate that EBV is not directly involved in the pathogenesis of PGL. PMID- 2544216 TI - The role of Epstein-Barr virus in lymphomas of HIV-carriers. AB - A substantial proportion of the lymphomas in HIV-carriers are EBV positive. Together with the fact that there are multiple signs of EBV-activation in AIDS patients and patients with ARC or PGL, this suggests that these virus carrying tumors develop as results of the immunosuppression, and that EBV has an important role in their pathogenesis. PMID- 2544217 TI - Effect of intravenously injected iodinated lipid emulsion on the liver. An experimental study correlating computed tomography findings with in vivo microscopy and electron microscopy findings. AB - Iodinated lipid emulsions have been shown to have great potential as site specific contrast media for the liver and spleen. Because of unacceptable adverse reactions none of these emulsions has been adopted for clinical use. In an attempt to find an explanation for these adverse reactions we tested three iodinated lipid emulsions, EOE-13, AG 60.99 and AG 66.18. The following models were used: Computed tomography (CT) of the rabbit liver, in vivo microscopy and electron microscopy of the rat liver. The emulsions contained particles of different sizes and were used in varying doses. We found that the larger the emulsion particles, the more likely they were to be taken up by the Kupffer cells and thereby the higher the opacification of the liver achieved at CT. We also observed changes in the microcirculation of the liver when the emulsions were given in doses required to secure satisfactory opacification of the liver at CT. The main changes were 1) a marked increase in the size of the Kupffer cells, and 2) damage to the sinusoidal endothelium, both contributing to sinusoidal congestion. These changes strongly suggest activation of the macrophages and this in turn probably results in the release of toxic mediators. We suspect that the adverse reactions observed in patients when using iodinated lipid emulsions are due to these toxic mediators. PMID- 2544218 TI - Expression of the small cell carcinoma antigens of cluster-5 and cluster-5A in primary lung tumours. AB - The expression of the small cell carcinoma (SCLC) antigens cluster-5 (antibody LAM8) and cluster-5A (antibody SWA20) was examined on a panel of routinely processed biopsy or surgical specimens of 290 lung tumours by immunoperoxidase staining. Antigen expression was largely restricted to SCLC. Of over 150 tissue samples evaluated, moderate or strong antigen expression was found in 49% (cluster-5) and 45% (cluster-5A). Concordance in expression of the two antigens was seen in 71% of SCLC samples, with 35% expressing both antigens strongly, 8% moderately and 28% being negative for both antigens. Antigen expression was independent of the morphological subtype of SCLC. Primary lung tumours of other histology, including squamous cell carcinoma, large cell carcinoma, adenocarcinoma, mesothelioma or carcinoid had no significant antigen expression. Of 135 tumours, strong or moderate expression of both antigens was seen only in two cases. 20%, mostly carcinoids, were weakly positive for cluster 5 and 4% for cluster 5A antigen. The remainder were antigen negative. No significant antigen expression was seen in 25 normal lung tissues. The membrane antigens of SCLC cluster 5 and 5A are markers for SCLC and their expression in tissues is tumour associated. PMID- 2544219 TI - Detection of human papillomavirus genes in human oral tissue biopsies and cultures by polymerase chain reaction. AB - We have used the polymerase chain reaction to detect DNA sequences related to human papillomavirus type 16, by simultaneous priming with oligonucleotides from the E6 and L1/L2 open reading frames of the HPV16 genome. The HPV16-related sequence is present at low levels in normal oral tissue, in addition to biopsies and cell cultures from patients with benign and malignant disease. Ultimate analysis of the amplified sequences from the E6(120bp) and L1/L2(173bp) regions of HPV16 was achieved by gel electrophoresis and comparative nucleotide sequencing. The oral carcinoma biopsies and tissue cultures contained DNA sequences which were identical to the E6 region of HPV16, but only rarely contained sequences closely related to the L1/L2 region. The PCR technology should permit the detection, identification and cloning of latent viruses from extremely small tissue biopsies. PMID- 2544220 TI - Expression of epidermal growth factor receptor (EGF-R) in non-small cell lung cancer. Use of archival tissue and correlation of EGF-R with histology, tumour size, node status and survival. AB - A total of 152 non-small cell lung cancers (NSCLC) were studied retrospectively to determine the relationship between epidermal growth factor receptor (EGF-R) status and the histological type, tumour size, nodal status and prognosis. EGF-R status was assessed on routinely embedded paraffin sections with an antibody to the cytoplasmic domain of the tumour (F4 antibody). EGF was demonstrated in all tumour types and every squamous and large cell carcinoma was positive for the antibody. Most tumours showed heterogeneity of staining. EGF expression was seen statistically more frequently in well differentiated tumours. Patients with 50% or more tumour cells showing positivity tended to have an improved survival but this result failed to reach statistical significance. There was no relationship between the size of the primary tumour or the lymph node status. Other cells, such as mucinous glands, bronchial epithelial cells and macrophages stained positively with the monoclonal antibody. EGF receptor status, with the antibodies presently available, adds little to help in either diagnosis or prognosis. Interpretation of data has to be guarded since the antibody was seen in some normal cells. PMID- 2544221 TI - Thyroid epithelial cell transformation by a retroviral vector expressing SV40 large T. AB - A recombinant murine retroviral vector encoding the SV40 virus large T antigen was used to infect stably an immortal line of differentiated rat thyroid epithelial cells, FRTL-5. Expression of SV40 T transformed these cells to anchorage independence and tumorigenicity but did not alter morphology or abolish tissue-specific functions and growth factor requirements. The resulting phenotype provides a model of well-differentiated human thyroid cancer. PMID- 2544222 TI - Patients at risk of chemotherapy-associated toxicity in small cell lung cancer. AB - During a clinical trial of duration of chemotherapy in small cell lung cancer (SCLC), 71 of 610 patients (11.6%) died in the first 3 weeks. Chemotherapy consisted of cyclophosphamide 1 g m-2 i.v. day 1, etoposide 100 mg t.d.s. orally days 1-3, vincristine 2 mg i.v. day 1. The time of death was found to be nonrandomly distributed within the first chemotherapy cycle, with a peak incidence between days 7 and 12 after chemotherapy. Patients were matched with controls who were the next cases entered into the study who did not die in the first 3 weeks. Patients dying early were more likely to have clinical hepatomegaly (P less than 0.0001), and ECOG score greater than or equal to 1 (P less than 0.00001). As a group these patients also had a higher alkaline phosphatase (P less than 0.0002), an elevated blood urea (P less than 0.00001) and a lower serum albumin (P less than 0.0001) than controls. It is probable that infection contributes to the death of these already ill patients at a time when the blood count is low. Early deaths have been noted in two other large trials using regimens including etoposide. Prophylactic antibiotics or dosage modification may prevent the early death of these high risk patients. PMID- 2544223 TI - A possible role for leukotriene B4 in head and neck cancer. PMID- 2544224 TI - The role of surgery in metastatic testicular germ cell tumours (GCT). PMID- 2544225 TI - Acute phase reaction during chemotherapy in small cell lung cancer. AB - We have measured the serum concentration of the acute phase reactant, C-reactive protein (CRP), in 20 patients with histologically proven small cell lung cancer undergoing their first pulse of induction cytotoxic chemotherapy. Baseline CRP concentrations were raised in 16 of 20 patients (median baseline CRP 18.5 mg l-1; normal range less than 10 mg l-1). CRP levels more than doubled in 11 of 20 patients during induction chemotherapy. This acute phase reaction was seen in seven of the 10 chemosensitive patients, but was not observed in any of the five non-responding patients. Five patients were non-evaluable for chemoresponse. These data indicate that there is a previously undescribed quantifiable acute phase response during chemotherapy for small cell lung cancer which has potential for predicting chemoresponse. PMID- 2544226 TI - IgM paraprotein reactive with muscle antigens associated with autonomic and peripheral neuropathy. PMID- 2544227 TI - Association of Tat protein and viral mRNA with nuclear matrix from HIV-1-infected H9 cells. AB - The transactivating protein from human immunodeficiency virus type 1 (HIV-1), Tat, was found to bind to the nuclear matrix from uninfected and HIV-1-infected H9 cells. Addition of the Zn2+, Cd2+ and Cu2+ chelator o-phenanthroline destroyed the matrix fibrils and the binding affinity of Tat to the matrix. A sequential treatment of the matrix, first with o-phenanthroline and then with ZnCl2, partially restored the fibrillar-like matrix structure. Infection of H9 cells with HIV-1 resulted in a displacement of cellular mRNA by viral mRNA from the nuclear matrix. Both the matrix-bound host cell and HIV-1 mRNA were found to dissociate from the matrix in the presence of o-phenanthroline. This could be prevented by coincubation with Zn2+ or Cu2+ (but not Mg2+), which stabilize the mRNA containing nuclear matrix structure. PMID- 2544228 TI - Inhibition of rat growth hormone promoter activity by site-specific DNA methylation. AB - The effect of methylation on rat growth hormone (rGH) promoter activity was determined in GH3 cells by measuring rGH-Neo and rGH-CAT fusion gene expression with or without prior in vitro treatment with the site-specific DNA methyltransferases, M-BsuE and M-HhaI. To assay for rGH-promoter-specific effects of DNA methylation, RSV-Neo and RSV-CAT activities with or without M-BsuE, M-HhaI and M-HpaII treatment were measured in parallel cultures of GH3 cells. GH1-Neo and RSV-Neo fusion gene expression was inhibited by in vitro methylation from 44 to 83% as measured by the number of Geneticin-resistant GH3 cell colonies. Methylation of the GH1 promoter by M-BsuE exhibited some selective inhibition of Neo expression as determined by colony numbers, although extensive methylation of non-promoter DNA in GH1-Neo and RSV-Neo by M-HhaI and M-HpaII also inhibited Neo expression. Southern blot analysis of genomic DNA isolated from the Geneticin resistant GH3 cells indicated that Geneticin-resistance was accompanied by demethylation of the BsuE (ThaI) sites in stably incorporated GH1-Neo DNA but not RSV-Neo DNA. Transient expression of the CAT gene in GH3 cells was selectively inhibited by 60% upon methylation of two BsuE (ThaI) sites in the GH1 promoter of GH1-CAT by M-BsuE. The data demonstrate, for the first time, to our knowledge, a direct effect of DNA methylation on the activity of the rat growth hormone promoter. PMID- 2544230 TI - Conformational transition of aquomethemoglobin: intramolecular histidine E7 binding reaction to the heme iron in the temperature range between 220 K and 295 K as seen by EPR and temperature-jump measurements. AB - Temperature-dependent EPR and temperature-jump measurements have been carried out, in order to examine the high-spin to low-spin transition of aquomethemogobin (pH 6.0). Relaxation rates and equilibrium constants could be determined as a function of temperature. As a reaction mechanism for the high-spin to low-spin transition, the binding of N epsilon of His E7 to the heme iron had been proposed; the same mechanism had been suggested for the ms-effect, found in temperature-jump experiments on aquomethemoglobin. A comparison of the thermodynamic quantities, deduced form the measurements in this paper, gives evidence that indeed the same reaction is investigated in both cases. Our results and most of the findings of earlier studies on the spin-state transitions of aquomethemoglobin, using susceptibility, optical, or EPR measurements, can be explained by the transition of methemoglobin with H2O as ligand (with high-spin state at all temperatures) and methemoglobin with ligand N epsilon of His E7 (with a low-spin ground state). Thermal fluctuations of large amplitude have to be postulated for the reaction to take place, so this reaction may be understood as a probe for the study of protein dynamics. PMID- 2544229 TI - EPR studies of the cytochrome-d complex of Escherichia coli. AB - We have examined the thermodynamic and EPR properties of one of the ubiquinol oxidase systems (the cytochrome d complex) of Escherichia coli, and have assigned the EPR-detectable signals to the optically identified cytochromes. The axial high spin g = 6.0 signal has been assigned to cytochrome d based on the physicochemical properties of this signal and those of the optically defined cytochrome d. A rhombic low spin species at gx,y,z = 1.85, 2.3, 2.5 exhibited similar properties but was present at only one-fifth the concentration of the axial high spin species. Both species have an Em7 of 260 mV and follow a -60 mV/pH unit dependence from pH 6 to 10. The rhombic high spin signal with gy,z = 5.5 and 6.3 has been assigned to cytochrome b-595. This component has an Em7 of 136 mV and follows a -30 mV/pH unit dependence from pH 6 to 10. Lastly, the low spin gz = 3.3 signal which titrates with an Em7 of 195 mV and follows a -40 mV/pH unit dependence from pH 6 to 10 has been assigned to cytochrome b-558. Spin quantitation of the high-spin signals indicates that cytochrome d and b-595 are present in approximately equal amounts. These observations are discussed in terms of the stoichiometry of the prosthetic groups and its implications on the mechanism of electron transport. PMID- 2544231 TI - Mitogen-stimulated lymphocyte proliferation and pituitary hormones in major depression. AB - To assess cellular immune status and the hypothalamic-pituitary (HP) axis in patients with major depression, we examined peripheral blood mononuclear cells (PBMC) and measured the plasma levels of cortisol, adrenocorticotropin hormone (ACTH), growth hormone (GH), and prolactin (PRL). Twenty patients with major depression were compared with 20 control subjects matched for age, sex, and race. The dose-response curves for concanavalin-A (Con-A) and phytohemagglutinin (PHA) stimulation were not significantly different between the two groups. The patients had decreased Con-A-stimulated T-lymphocyte proliferation when compared to the control subjects, but only at the lowest suboptimal concentration of Con-A. None of the four concentrations of PHA-stimulated proliferation were different between the two groups, neither was PHA-induced interleukin-2 production. Within the patient group only, plasma prolactin (PRL) correlated significantly with stimulated lymphocyte proliferation using two optimal concentrations of PHA and one optimal concentration of Con-A, when the proliferation was expressed using the stimulation index. PMID- 2544232 TI - Risk lesions in cirrhosis and development of hepatocellular carcinoma: an autopsy study. AB - Non-neoplastic morphologic changes in various types of cirrhosis were evaluated in relationship to the presence or absence of hepatocellular carcinoma (HCC), using autopsy livers from Hokuriku (Japan) and Los Angeles (USA). Macronodular cirrhosis was closely related to HCC in B-viral cirrhosis, alcoholic cirrhosis and cirrhosis of uncertain type. Liver cell dysplasia was most frequently seen in cases with and without HCC in B-viral cirrhosis but was significantly more frequent with HCC in cases of alcoholic cirrhosis and cirrhosis of uncertain type. Nodular bulging activity within regenerative nodules was closely related to HCC in alcoholic cirrhosis. A positive relationship between HCC and Mallory bodies was found in non-alcoholic cirrhosis. These data suggest that patients with macronodular cirrhosis, liver cell dysplasia, nodular bulging activity and Mallory bodies may have an increased risk of developing, or having HCC dependent on the etiology of cirrhosis. The geography and race differences had some relationship to the incidence of HCC. PMID- 2544233 TI - [The development of ischemic cardiac arrhythmias during naloxone blockade of the opiate receptors]. AB - Acute experiments on cats have shown that the naloxone blocks of opiate receptors increased essentially the incidence of idioventricular arrhythmias in myocardial ischemia. These results may evidence of the endogenous opioid peptides involvement in the body response on the acute myocardial ischemia. PMID- 2544234 TI - [The content of cyclic nucleotides in an organ culture of normal and atherosclerosis-affected human aorta]. AB - Cyclic AMP and cyclic GMP content was measured in intima media of unaffected and atherosclerotic areas of human aorta in a short-term organ culture. It was demonstrated that during short-term cultivation the content of both cyclic nucleotides in tissues is constant. The cyclic AMP content in fatty streaks and atherosclerotic plaques is significantly (2 to 7-fold) lower than in unaffected intima. The cyclic GMP level in atherosclerotic lesions is 1.5 to 3-fold higher than in normal. The content of both cyclic nucleotides in the media underlying fatty streaks and atherosclerotic plaques is the same as in the normal tissue. The obtained data indicate serious disorders in the system of cyclic nucleotides during atherosclerosis. PMID- 2544235 TI - [The measurement of the amount of iron (III) complexes with desferal in the perfused rat liver by an EPR method]. AB - Rat liver was perfused by Hank's solution, containing desferal (deferoxamine). It was shown that in perfusion of the liver spectrum EPR a signal (g = 4.3; H = 63 G) appears. This signal belongs to desferal complexes, containing intracellular Fe/3/. Desferal transfer to the liver tissue and further formation of desferal complexes there takes place within first 5-10 min of liver perfusion by solution, containing 0.5 mM of desferal. PMID- 2544236 TI - [Activation of the fast calcium input in the denervated smooth muscle of the cat nictitating membrane]. AB - The excitation and contraction features of innervated and sympathetically denervated smooth muscle strips from cat's nictitating membrane have been studied by single sucrose gap arrangement. Increasing of smooth muscle cells sensitivity to drugs were accompanied by elevation of membrane response and the ability to generation of action potentials. Action potentials have been induced by agonists or high potassium concentration in external solution and spontaneously. In innervated muscle action potentials have been evoked as a result of depolarization by high potassium concentration of TEA blockade of potassium conductance. Induced and spontaneously generated action potentials were blocked by organic and inorganic antagonists of potential dependent Ca++ channels. In Ca free solution action potentials were absent but might be supported by Ba++. Decrease of Na+ had no effect on smooth muscle excitability. It is supposed that activation of potential depended Ca++ channels in smooth muscle cells with pharmaco-mechanical coupling are under influence of sympathetic nerves. PMID- 2544237 TI - [Electron microscopic study of the cytoskeleton of human podocytes]. AB - The ultrastructural study of man's cytoskeleton of podocytes is carried out. Populations of podocytes with two different types of structure of the cytoskeleton in dependence on age (2, 4, 6, 37 and 65 years) is revealed in kidneys. The first type of cytoskeleton of the podocyte is peculiar for children's age and is characterized by branched, high density microfilament network, expressed by system of microtubules and single myofilaments. The intermediate filaments here are either utterly absent or present so feebly they find themselves "disguised" by other strongly developed components of cytoskeleton and revealing them with the help of technique of electron microscope is impossible. In kidneys of adults, and especially of old aged persons podocytes with other type of organization of the cytoskeleton are mainly identified. The distinctive signs of the last are bundle arrangement of microfilaments, plural bundles of intermediate filaments and individual microtubules. This study permits to make a conclusion that during individual development and growing old in kidneys of high animals and man, probably, physiological changes causing morphological reconstruction of cytoskeleton which is accompanied by intensive development of intermediate filaments' system with simultaneous "involution" of microtubules and microfilaments' systems take place. PMID- 2544238 TI - [Primary brain tumors in children. I]. AB - The most frequent solid tumors in the pediatric age group are central nervous system tumors. Despite dramatic improvements in neurosurgery, radiotherapy and chemotherapy, the overall prognosis remains poor, and is often associated with late deleterious effects, including neuropsychologic disorders, endocrine deficiencies and physical abnormalities. In part I, we describe the common presenting symptoms and signs in children with brain tumors. We expose the significant advances in histology, immunohistochemistry, cytogenetic, radiology and therapeutic approaches during the past ten years. PMID- 2544239 TI - Characterization of insulin-like growth factor 1 receptors (IGF1-R) in human breast cancer cell lines. AB - The binding characteristics of IGF1 on membranes prepared from 5 human breast cancer cell lines were investigated in detail. The presence of one class of high affinity IGF1 binding sites was demonstrated (BT-20: n = 230 fmol/mg protein, Ka = 0.7 nM-1; MCF-7: n = 124 fmol/mg protein, Ka = 1 nM-1; T-47D: n = 61 fmol/mg protein, Ka = 1.1 nM-1; HBL-100: n = 18 fmol/mg protein, Ka = 3.2 nM-1; MDA-MB 231: n = 7 fmol/mg protein, Ka = 2.8 nM-1). Chemical cross-linking of 125I IGF1 to breast cancer cell membranes then sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed one major band of relative molecular weight 130000. The specificity of these receptors was studied: native or recombinant IGF1 had the same potency to inhibit 125I IGF1 binding; IGF2 was able to compete for binding, whereas insulin competed with a potency lower than 1/100 that of IGF1. These characteristics of IGF1 binding sites in breast cancer cell membranes correspond to the previously described binding unit of type I IGF receptors (IGF1-R). Finally we determined that for a routinely used standard assay, it was necessary to incubate for 5 h at 4 degrees C a high amount of membrane protein (400 micrograms) and 200,000 cpm of tracer. Considering the known effect of IGF1 on breast cancer cell multiplication, it is tempting to suggest that this factor might play a major role in the growth of breast cancer: the measurement of IGF1 R, using this standardized method, will give an assessment of these tumors IGF1 sensitivity; it can be performed on the membrane fraction obtained when preparing cytosol for steroid receptor assay. PMID- 2544240 TI - Electronic interaction chromatography on porous graphitic carbon. Separation of [99mTc]pertechnetate and perrhenate anions. AB - The oxo anions of technetium-99m and rhenium, pertechnetate (TcO4-) and perrhenate (ReO4-), have been separated by high performance liquid chromatography on porous graphitic carbon with aqueous trifluoroacetic acid or salt solutions as eluent. Chromatographic retention was a result of electronic interaction between the lone pair electrons of the anions and the delocalised pi-electron clouds of the porous graphitic carbon. Retention and resolution can be controlled by the concentration and/or species of eluents which can compete with the solutes for electronic interaction being used for elution. PMID- 2544241 TI - Fluconazole and phenytoin: a predictable interaction. PMID- 2544242 TI - Upper-half body irradiation for oat cell carcinoma of the bronchus. PMID- 2544243 TI - Continuous single-layer gastrointestinal anastomosis: a prospective audit. AB - Single-layer bowel anastomoses have conventionally been constructed using an interrupted suture technique. A single-layer continuous technique has been avoided on the grounds that it may predispose to ischaemia of the bowel ends. We have routinely used a single-layer continuous suture technique with an absorbable suture material (polyglycolic acid) to construct all recent intraperitoneal bowel anastomoses, and we present a 3-year audit of this technique. A total of 131 patients were studied of whom 66 had undergone upper gastrointestinal resections and 65 had had colonic resections. Twenty-two patients had emergency operations. Anastomotic failure was noted in 4.5 and 6.2 per cent of patients in each group respectively. The incidence of minor wound infection was 1.5 and 7.7 per cent respectively. The overall mortality rate was 8.4 per cent. This study suggests that a single-layer continuous suture technique gives acceptably reliable results when used in gastrointestinal anastomosis. PMID- 2544244 TI - Low molecular weight heparins and prevention of postoperative thrombosis. PMID- 2544245 TI - Phencyclidine and MK-801: a behavioral and neurochemical comparison of their interactions with dihydropyridine calcium antagonists. AB - The impairment of rotarod ability and the convulsive activity of phencyclidine (PCP) and MK-801 were compared in male CD-1 mice. The putative interaction between nifedipine and PCP and MK-801 on these behavioral measurements was also quantitated and compared. MK-801 produced a dose dependent inhibition of rotarod ability with an ED50 of 0.5 mg/kg. Nifedipine potentiated the impairment of rotarod ability by MK-801. Both PCP and MK-801 produced convulsive behavior in mice which was characterized by jumping and wild running fits; the CD50 for MK 801 was 1.3 mg/kg. Nifedipine dose dependently inhibited the convulsions associated with MK-801 and PCP. PCP but not MK-801 increased [3H]nitrendipine binding to dihydropyridine (DHP) binding sites on mouse brain membranes. MK-801 blocked the effects of PCP on [3H]nitrendipine binding. These findings suggest that MK-801 is a potent PCP-like drug which interacts with nifedipine and neuronal DHP binding sites. Nifedipine's reduction of the hyperactivity and convulsions elicited by MK-801 may be of importance in the eventual development of MK-801 as an antiischaemic and anticonvulsant drug. PMID- 2544246 TI - Locus coeruleus potentiation of dentate gyrus responses: evidence for two systems. AB - Glutamate activation of the locus coeruleus (LC) and norepinephrine (NE) have both been shown to potentiate the perforant path (PP)-evoked population spike. This potentiation may be short-lasting, the population spike returning to baseline levels within minutes after NE-application or LC activation, or can be long-lasting, persisting 20 minutes or more after termination of the NE or glutamate manipulation. In the present study LC electrical stimulation (333 Hz, 15 msec) initiated 40 msec prior to a PP stimulus reliably caused short-lasting potentiation of the dentate gyrus population spike amplitude (mean maximal = 161%, N = 22). With 50 LC-PP pairings a long-lasting potentiation (greater than 30 min after offset of LC stimulation) was seen in 10/22 experiments. Propranolol (20-30 mg/kg IP) did not block the potentiating effect of LC electrical simulation but completely suppressed the potentiating effect of glutamate activation of the LC in the same animals (N = 5). The beta receptor dependence of short-and long-lasting hippocampal NE potentiation has been previously demonstrated. The inability of a beta receptor antagonist to attenuate the potentiation induced by LC electrical stimulation suggests there are two distinct systems. Both the beta-NE-dependent and the beta-NE-independent system are capable of inducing long-lasting potentiation of the PP-evoked potential. PMID- 2544247 TI - Hepatic perfusion with FUdR utilizing an implantable system in patients with liver primary cancer or metastatic cancer confined to the liver. AB - Nineteen patients with colorectal adenocarcinoma, three with cholangiocarcinoma, two with hepatocellular carcinoma, and one with carcinoid were treated with hepatic artery infusion chemotherapy. An implantable pump system was used to deliver floxuridine (FUdR), starting at 400 mg for 2 weeks with 2 weeks of rest. Eleven of 15 (73%) measurable patients with colorectal carcinoma responded. Of 6 complete responses, 4 were documented by laparotomy, including 1 with cholangiocarcinoma. Toxicity included dyspepsia and elevated liver function tests in all patients, gastric ulcer in 2, cholecystitis in 2, and sclerosing cholangitis in 3. Overall median survival for the colon cancer patients has not been reached at 16 months. Regional disease was controlled in the majority of patients treated with this regimen with acceptable toxicity and good quality of life. PMID- 2544248 TI - Nedocromil sodium is more potent than sodium cromoglycate against AMP-induced bronchoconstriction in atopic asthmatic subjects. AB - Nedocromil sodium is a new chemical entity which shows similar properties to sodium cromoglycate (SCG) and in addition exhibits a preferential activity in stabilizing mucosal mast cells. We have compared the effect of inhalation of nebulized placebo, SCG and nedocromil sodium on the bronchoconstrictor response to inhaled adenosine monophosphate (AMP) in eight atopic asthmatic subjects aged 25 yr (range 21-32 yr). The geometric mean provocation doses of AMP required to produce a 20% decrease in FEV1 (PD20FEV1) and a 40% decrease in Vmax30 (PD40 Vmax30) following placebo were 4.9 (0.3-14.2) and 1.8 (0.1-8.4) mumol respectively. Prior inhalation of both SCG and nedocromil sodium significantly inhibited the bronchoconstrictor response to AMP with PD20FEV1s of 36.6 (4.0 132.7) and 134 (12.4-560), and PD40 Vmax30 values of 20.5 (1.4-110) and 101.6 (5 560) mumol respectively (P less than 0.001). Nedocromil sodium was 3.9 (FEV1) and 8.0 (Vmax30) times more potent than SCG (P less than 0.001). In conclusion, both drugs inhibit the bronchoconstrictor response to inhaled AMP, and nedocromil is at least 4-8 times more potent than SCG. PMID- 2544249 TI - Treatment of Wilms' tumor. Results of the Third National Wilms' Tumor Study. AB - The Third National Wilms' Tumor Study sought to reduce treatment for low-risk patients and find better chemotherapy for those at high risk for relapse. Eligible patients (1439) were randomized according to stage (I-IV) and histology (favorable [FH] or unfavorable [UH]), and contributed data to survival and relapse-free survival (RFS) analyses. Four-year (postnephrectomy) survival percentages and randomized treatment regimens for low-risk patients were 96.5% for 607 Stage I/FH patients who received dactinomycin (Actinomycin D [AMD], Merck Sharp & Dohme, West Point, PA) and vincristine (VCR) for 10 weeks versus 6 months; 92.2% for 278 Stage II/FH patients; and 86.9% for 275 Stage III/FH patients who received AMD + VCR +/- Adriamycin (ADR, Adria Laboratories, Columbus, OH) for 15 months. Stage II/FH patients also had either zero or 2000 cGy irradiation (RT) postoperatively and Stage III/FH patients either 1000 or 2000 cGy. Four-year survival was 73.0% for 279 high-risk patients (any Stage IV, all UH) who received postoperative radiation therapy (RT) and AMD + VCR + ADR +/- cyclophosphamide (CPM). Statistical analysis of survival and RFS experience shows that the less intensive therapy does not worsen results for low-risk patients and CPM does not benefit those at high risk. PMID- 2544250 TI - Increased lipid peroxidation in malignant tissues of patients with colorectal cancer. AB - The authors investigated the level of lipid peroxidation and possible related phenomena in the activation of phospholipase A2 and the infiltration of granulocyte neutrophils in human colorectal cancer tissue samples. Malondialdehyde (an index of lipid peroxidation), phospholipase A2 activity, and myeloperoxidase activity (a marker of granulocyte neutrophils) were found to be increased significantly in cancerous compared with macroscopically normal tissues. They concluded that increased lipid peroxidation, phospholipase A2, and myeloperoxidase activity are associated with human colorectal cancer. PMID- 2544251 TI - Influence of infiltrating lobular histology on local tumor control in breast cancer patients treated with conservative surgery and radiotherapy. AB - To determine the influence of infiltrating lobular histology on local tumor control, the authors studied 49 patients with Stages I and II infiltrating lobular breast carcinoma treated by limited excision of the tumor and radiotherapy between 1968 and 1981 (median follow-up, 75 months). Results were compared with those in 561 cases of infiltrating ductal carcinoma similarly treated during the same period. The 5-year actuarial risk of local recurrence was similar for patients with infiltrating lobular or ductal carcinoma when the latter was evaluated as a single group (12% versus 11%). However, the 12% 5-year actuarial local recurrence risk for patients with infiltrating lobular carcinoma was intermediate between that for patients with infiltrating ductal carcinomas with an extensive intraductal component (23%) and those without an extensive intraductal component (5%). The pattern of recurrence in the breast was similar in the infiltrating lobular and ductal groups. All recurrences in patients with infiltrating lobular carcinoma and 80% of recurrences in the infiltrating ductal group occurred in the vicinity of the primary tumor (P = not significant). None of the clinical or morphologic features examined significantly influenced the risk of local recurrence in patients with infiltrating lobular carcinoma. The authors conclude that combined conservative surgery and radiotherapy appear to be a reasonable treatment option for patients with infiltrating lobular carcinoma, but further follow-up will be required to confirm these results. PMID- 2544252 TI - The role of morphometry in predicting prognosis in pancreatic islet cell tumors. AB - Morphometry of 31 pancreatic islet cell tumors was examined to determine the value of this technique in assessing tumor behavior. Patients were followed for a mean period of 5.1 years (range, 1 month-14 years) after diagnosis. Initially 17 localized and nine metastatic tumors were studied. Discriminant analysis was carried out on these cases and identified nuclear/cytoplasmic ratio and number of nuclei/mm2 as the significant discriminatory features. These were combined to derive a classification rule which was capable of correctly identifying localized and metastatic tumors in 92% of cases. The classification rule was applied subsequently to an additional five test cases, all of which were classified successfully. The failure of increased nuclear size and pleomorphism to correlate with malignancy in these tumors was confirmed. Tumors which metastasized had significantly greater gross diameters than localized lesions, but overlap existed. Mitotic counts were not a helpful discriminatory feature. Morphometry may be useful in improving histologic assessment of pancreatic islet cell tumor behavior. PMID- 2544253 TI - Multilocular cyst of the kidney (cystic nephroma) and cystic, partially differentiated nephroblastoma. Terminology and criteria for diagnosis. AB - The entities commonly known as multi-locular cyst of the kidney (MLC) and cystic partially differentiated nephroblastoma (CPDN) were reviewed, based on material in the National Wilms' Tumor Study Pathology Center. The authors recommend several modifications of existing terminology and definitional criteria for these lesions. Because MLC probably represents a neoplastic lesion, the designation "cystic nephroma" (CN) is preferred. This term should be used only for predominantly cystic tumors composed entirely of differentiated tissues, without blastema or other embryonal elements. The designation CPDN should be applied to predominantly cystic lesions, lacking nodular solid regions, in which blastemal or other embryonal cells are present in the septa of the cysts. Solid Wilms' tumor with multifocal cystic change should be distinguished from CPDN. Five cases of CN and 18 cases of CPDN were reviewed. No CN, for which follow-up data was available, showed aggressive behavior. Only one case of CPDN underwent local recurrence, and there were no metastases. In general, nephrectomy alone appears to be adequate therapy for CPDN, but regular monitoring by noninvasive techniques would seem advisable. PMID- 2544254 TI - Integration of nutrition support into oncologic treatment protocols for high and low nutritional risk children with Wilms' tumor. A prospective randomized study. AB - Benefits and risks of nutrition support were evaluated in 31 malnourished children with newly diagnosed Wilms' tumor managed according to the third National Wilms' Tumor Study protocol. Patients were classified at diagnosis as being at high nutritional risk (HNR, n = 19) or low nutritional risk (LNR, n = 12). Ten HNR patients were randomized to central parenteral nutrition (CPN) and nine HNR patients were randomized to peripheral parenteral nutrition (PPN) plus enteral nutrition (EN) for 4 weeks of initial intense treatment and EN (nutritional counseling, oral foods and supplements) thereafter. Thirteen HNR patients (seven CPN, six PPN) completed the protocol. Twelve LNR patients received EN; 11 Stage I malnourished patients were randomized to 10 or 26 weeks of chemotherapy. Dietary, anthropometric, and biochemical data were determined for HNR patients at weeks 0-4, 6, 13, 19, and 26 and for LNR patients at weeks 1, 2, 5, and 26. In HNR patients, adequate parenteral nutrition support reversed protein energy malnutrition (PEM), and prevented chemotherapy and radiotherapy delays due to granulocytopenia. CPN was superior to PPN in reversing PEM: energy intake, weight gain, and retinol binding protein were higher (P less than 0.05). LNR patients lost weight and fat reserves in the first 2 weeks of treatment; depletion persisted at week 5, and 25% had chemotherapy delays. Thereafter, EN reversed PEM in patients with both chemotherapy regimens. These data suggest that CPN is preferable during initial intense treatment for HNR patients, and that, although EN is ineffective in preventing depletion and treatment delays in the first 5 weeks of treatment for LNR patients, it is effective thereafter. PMID- 2544255 TI - Surgery of small hepatocellular carcinoma. Analysis of 144 cases. AB - A long-term follow-up study of 144 cases with surgically and pathologically proved small hepatocellular carcinoma (less than or equal to 5 cm) from 1967 to 1987 is reported. One hundred eight cases (75.0%) were detected by alpha fetoprotein serosurvey and/or ultrasonography mainly in a high-risk population; 129 cases (89.6%) coexisted with cirrhosis. Resection was done in 132 cases (91.7%) with three (2.3%) operative deaths; cryosurgery, laser vaporization, and hepatic arterial chemotherapy were used in the rest. Limited resection was done in 67.4% of resections. Reresection of subclinical recurrence or solitary pulmonary metastasis was done in 21 cases. The 5-year and 10-year survival rates were 67.9% and 53.4% in the resection group but zero in the nonresection group. Survival was correlated negatively with tumor size, 5-year survival after resection was 84.6% in tumors less than or equal to 2 cm but 59.5% in tumors of 4.1 to 5 cm. The increase of resectability and reresection resulted in marked improved of 5-year survival from 43.5% in 1973 to 1977 to 63.3% in 1978 to 1982 in the entire series. No significant difference was found between survival of limited resection and lobectomy. Resection may be the modality of choice for treatment of small hepatocellular carcinomas with compensated liver function. Limited resection instead of lobectomy was the key to increased resectability and decreased operative mortality in cirrhotic livers. Reresection of subclinical recurrence was important to prolong survival further. PMID- 2544256 TI - Cholangitis, hemobilia, and cholangiocarcinoma. Management of a fistula between an obstructed right hepatic duct and the portal vein. AB - Cholangitis is the most common cause of sepsis in patients with obstructing carcinomas of the biliary tree. Catheter and stent placement may relieve or exacerbate the septic course. Transhepatic stent placement produced a cholangioportal fistula and hemobilia in a patient with cholangiocarcinoma. The described technique of retrograde operative decompression and clot evacuation may be used in patients in whom portal dissection is hazardous or in whom preservation of an existing cholangiojejunostomy in the portal region is desired. PMID- 2544257 TI - Human papillomavirus (HPV)-associated male and female genital carcinomas in a Hindu population. The male as vector and victim. AB - Epidemiologic, histologic and immunohistochemical data concerning male (penile) and female (cervical and vulvar/vaginal) genital carcinomas in a Hindu population are reported. The data are from Bali, an Indonesian Hindu island in a country with a predominantly Muslim population. In contrast to the surrounding Muslim population, circumcision is rare in the people of Bali, and the rate of phimosis in grown men is very high. The Balinese epidemiologic data of 1985 to 1986 were compared with 1986 data from The Netherlands. In Bali, cervical carcinoma was the most frequent carcinoma in women, and vulvar/vaginal carcinoma ranked seven. These carcinomas were much less frequent in The Netherlands. In Balinese men, penile carcinoma is the second most frequent carcinoma, whereas it is rarely diagnosed in The Netherlands. Penile carcinomas are found in Bali in younger age groups than in The Netherlands. Based on the immunohistochemical staining results, it is estimated that over 75% of the studied Balinese genital carcinomas contain human papillomavirus (HPV). The data presented in this paper may indicate that the cofactor of impeded postcoital hygiene can be of great importance for male and female HPV-associated genital carcinogenesis. It is clear that Balinese men, in particular men with extreme phimosis, are both vectors and victims of HPV. In The Netherlands the man is exclusively the vector of HPV. PMID- 2544258 TI - Cysteine protease activities and tumor development in human colorectal carcinoma. AB - Many studies of malignant cells or tissues in culture have implicated cysteine proteases in the progression of malignancy. We have extended these observations by measuring quantitative and qualitative changes in the expression of cathepsin B-like and L-like cysteine proteases during the growth and development of human colorectal carcinomas. Data derived from matched pairs of normal colorectal mucosa and carcinoma tissue from 27 patients demonstrated that both cathepsin B like and cathepsin L-like specific activities were significantly elevated (P less than 0.005) in the carcinoma tissue, while levels of endogenous cysteine protease inhibitor remained constant. Correlation of cathepsin enzyme activities with different stages of colorectal cancer demonstrated significantly higher cysteine protease activities in individuals with Dukes' A tumors (tumors confined to the bowel wall) than in patients with more advanced tumors (Dukes' B, C, or D tumors) (P less than 0.01-0.05). The relative proportion of activities contained in tumor epithelial and stromal elements remains to be elucidated. These results suggest an important role for cysteine proteases in the early progression of human colorectal carcinoma. PMID- 2544259 TI - Differential induction of spermidine/spermine N1-acetyltransferase in human lung cancer cells by the bis(ethyl)polyamine analogues. AB - We have investigated the induction of an important polyamine metabolic enzyme, spermidine/spermine N1-acetyltransferase, in two human lung cancer cell lines which respond differently to treatment with the bis(ethyl)polyamine analogues. The human small cell lung carcinoma line NCI H82 has previously been shown to be minimally affected by treatment with these analogues, whereas the large cell undifferentiated lung carcinoma line, NCI H157, responds in a rapid cytotoxic manner (R.A. Casero, Jr., S. J. Ervin, P. Celano, S. B. Baylin, and R. J. Bergeron, Cancer Res., 49:639-643, 1989). The mechanisms underlying the differential response are unknown. In the responsive NCI H157 cells, the bis(ethyl)polyamines were found to induce spermidine/spermine N1 acetyltransferase in a time- and dose-dependent manner to maximum levels greater than 1700-fold over baseline. By contrast, the unresponsive NCI H82 cells exhibit minimal induction of spermidine/spermine N1-acetyltransferase to less than 7-fold increase after bis(ethyl)polyamine treatment, regardless of time or concentration examined. The results of the current study suggest that the differential induction of this key enzyme, which is rate limiting in the back conversion pathway of polyamine metabolism, may play a role in determining cell specific to the bis(ethyl)polyamine analogues. PMID- 2544260 TI - Adriamycin activation and oxygen free radical formation in human breast tumor cells: protective role of glutathione peroxidase in adriamycin resistance. AB - Previous studies with Adriamycin-sensitive and -resistant (ADRR) MCF-7 human breast tumor cell lines indicated that Adriamycin formed significantly less hydroxyl radical (.OH) as the result of enhanced detoxification of reactive oxygen intermediates in the ADRR cell line. In order to further define the sites of drug activation and the role of detoxification mechanisms in free radical levels, subcellular fractions were isolated from these two cell lines and free radical formation in the presence of Adriamycin was examined by using electron spin resonance spectroscopy. Studies reported here show that considerable NADPH cytochrome P-450 reductase and NADH dehydrogenase activities were present in microsomes and mitochondria, respectively, and in nuclei obtained from these cells, and the relative activity of NADH dehydrogenase was 2-fold higher in the mitochondrial fraction of ADRR cells compared to the mitochondrial fraction from the parental wild type cells. In the presence of Adriamycin and a reducing cofactor (NADPH or NADH), Adriamycin semiquinone free radical, superoxide anion, and .OH were detected in all these fractions. Although only a small difference in the relative amount of oxy radical formation was detected in tumor microsomes, both mitochondria and nuclei of ADRR cells showed an overall 2-fold decreased formation of oxy radicals. The formation of the free radicals was significantly inhibited by superoxide dismutase, catalase, and dimethyl sulfoxide, indicating that free .OH generation was both superoxide and hydrogen peroxide dependent. The addition of purified glutathione peroxidase likewise inhibited .OH formation in a dose-dependent fashion. Similarly, when the lysate from ADRR cells, which contains 12- to 14-fold more glutathione peroxidase than Adriamycin-sensitive cells, was added to reaction mixtures containing Adriamycin-sensitive cells and Adriamycin, the .OH formation was diminished. Decreased free radical formation in nuclei and mitochondria, as a result of detoxification of hydrogen peroxide by glutathione peroxidase, may be significant in the protection of ADRR cells from Adriamycin-induced cell killing. PMID- 2544261 TI - Secretion of parathyroid hormone-like activity from human T-cell lymphotropic virus type I-infected lymphocytes. AB - Because many patients with adult T-cell leukemia/lymphoma (ATLL) develop hypercalcemia with similar characteristics to those of humoral hypercalcemia of malignancy (HHM) (Arch. Intern. Med., 148: 921-925, 1988), we investigated if ATLL cells produce parathyroid hormone (PTH)-like activity. Conditioned media from cultures of human T-cell lymphotropic virus type I-infected cell line (MT-2) as well as peripheral lymphocytes from a hypercalcemic ATLL patient stimulated cyclic AMP production in osteoblast-like rat osteogenic sarcoma cells (UMR 106) and bone resportion in organ cultures of fetal mouse calvaria. Furthermore, the stimulation of cyclic AMP production by conditioned medium of MT-2 cells was inhibited by human PTH(3-34), indicating that MT-2 cells secrete PTH-like activity. The PTH-like activity from MT-2 cells was chromatographically indistinguishable from the one extracted from a solid tumor causing HHM. The present results along with our previous observation that MT-2 cells constitutively express mRNA for PTH-related protein (Biochem. Biophys. Res. Commun., 154: 1182-1188, 1988) demonstrate that a PTH-like activity is synthesized and secreted by these cells, and are consistent with the hypothesis that elaboration of PTH-like activity by ATLL cells may be the mechanism by which hypercalcemia develops in ATLL patients as well as in solid cancer patients with HHM. However, these results do not rule out the possibility that other factors such as interleukin 1 are also involved and may act in concert with PTH-like activity in the development of hypercalcemia in ATLL. PMID- 2544262 TI - Characterization of sequences related to the mouse mammary tumor virus that are specific to MCF-7 breast cancer cells. AB - Mouse mammary tumour virus (MuMTV) DNA hybridized more strongly to two human EcoRI fragments (6.6 and 9.5 kilobases) in DNA from the MCF-7 human breast cancer cell line than in DNA from normal human placenta. Seven recombinants (NMWV 1E1-7) containing these MuMTV-related sequences were identified. Hybridization of NMWV 1E probes to Southern transfers of human DNA suggested that these probes hybridize to multiple sequences in the human genome. The pattern obtained was very similar to that obtained using MuMTV gag-pol DNA. Analysis of the cloned DNA showed that the NMWV 1E MuMTV-related sequences are arranged as tandem repeats and are contained in EcoRI fragments of 6.6 or 9.5 kilobases. Only two NMWV 1E EcoRI fragments (9.5 and 15 kilobases) were detected in 17 DNA samples prepared from human placenta and blood. In contrast the 6.6-kilobase EcoRI fragment was detected in two (MCF-7 and EFM-19) of seven breast cancer cell lines and the MDA MB-231 cell line contained NMWV 1E sequences in an EcoRI fragment of 9.8 kilobases. PMID- 2544263 TI - Characterization of the 3' region of the human DNA topoisomerase I gene. AB - Previous studies suggest that topoisomerase I (Topo I) plays a critical role in cell growth. However, the structure of the Topo I gene has not yet been determined. Two complementary DNA (cDNA) clones for the human Topo I 4.1-kilobase mRNA were isolated independently from HeLa and KB cell cDNA libraries. These clones were identical and they contained 679 base pairs of coding and 1138 base pairs of noncoding sequences. The clones had a two-base difference in the 3' noncoding region compared to the Topo I cDNA from human placenta. The structure of the 3' end of the human Topo I gene from six human tumor cell lines was examined. The Topo I cDNA recognized 16.5, 24.2, and 16.0 kilobases of genomic DNA restricted with EcoRI, HindIII and PstI, respectively. The individual genomic fragments were ordered by double digestion and hybridization with cDNA subclones. digestion and hybridization with cDNA subclones. The results indicate that the human Topo I gene contains several intervening sequences. The gene arrangement was similar in all six cell lines and no polymorphism was observed. However, each digestion contained genomic fragments that hybridized with all the subclones, suggesting that at least one Topo I pseudogene, or another Topo I gene with a different structure, was present in every cell line. As predicted, double digestions generated at 161 base pair fragment that indicates the presence of an intronless pseudogene. In contrast to the DNA topoisomerase I gene, the presumptive pseudogene(s) appears to be hypomethylated. In addition to the 4.1 kilobase Topo I mRNA, a larger 6-kilobase RNA was identified in human KB and HeLa cells which could be a processed Topo I mRNA intermediate. PMID- 2544264 TI - Purification and some properties of a lung-derived growth factor that differentially stimulates the growth of tumor cells metastatic to the lung. AB - The ability of malignant cells to respond to growth factor(s) present in or secreted by a distant target organ may be important in tumor metastasis. We used metastatic cell lines and clones of the rat 13762NF mammary adenocarcinoma that show reproducible spontaneous metastatic behavior from the mammary fat pad to regional lymph nodes and lung sites. Whereas poorly lung metastatic MTPa and MTC cells did not grow in response to lung-conditioned medium, highly lung-metastatic MTLn3 cells responded and grew rapidly in lung-conditioned medium. The major growth-promoting factor for MTLn3 cells from porcine and rat lung-conditioned media was purified by using hydroxylapatite affinity and anion exchange chromatography, chromatofocusing, size exclusion chromatography, and preparative native gel electrophoresis. The activity in each of the purification fractions was measured by determining their ability to increase the number of MTLn3 cells in serum-deprived culture. The major component that differentially stimulated the growth of highly metastatic MTLn3 cells was a glycoprotein of Mr approximately 66,000. Under reducing conditions, its apparent Mr was approximately 72,000. This lung-derived mitogen was stable at pH 4.0-9.0, possessed a pI of 6.9-7.0, and preferentially promoted the growth of lung-metastasizing tumor lines over their poorly lung-metastasizing counterparts in rat 13762NF mammary adenocarcinoma and murine B16 melanoma tumor systems. The activity of porcine lung-derived growth factor was not affected by pretreatment with antisera to porcine insulin, human granulocyte-macrophage colony-stimulating factor, human platelet-derived growth factor, or murine epidermal growth factor. It was inactivated by reduction with dithiothreitol or exposure to high temperature (95 degrees C). The results suggest that specific organ-derived growth factors are important in metastatic colonization and organ growth of particular malignant cells. PMID- 2544265 TI - Reduced cerebrospinal fluid B-endorphin levels in Rett syndrome. AB - Cerebrospinal fluid (CSF) levels of B-endorphin (B-EP), B-lipotropin (B-LPH) and ACTH were measured in nine girls with Rett syndrome with features of autistic behavior (3.7-12.1 years of age) and in ten children with chronic leukemia (control group). The peptides were measured by radioimmunoassay, either directly in the sample (ACTH) or after Sephadex G-75 column chromatography, in order to eliminate interfering substances (B-LPH and B-EP). The CSF B-EP patient levels (20.8 +/- 13.1 fmol/ml, means +/- SD) were significantly lower than in age matched controls (69.1 +/- 32.6, P less than 0.01), whereas the B-LPH and ACTH levels were in the control range. No correlations were found between the clinical findings and CSF neuropeptide concentrations. These data demonstrate a decrease in central opiate activity in girls with Rett syndrome. PMID- 2544266 TI - Interventional angiography in neuropediatrics. AB - Since the introduction of non-invasive imaging techniques (CT, US, MRI), superselective cerebrospinal angiography has been playing a major role as a diagnostic tool as well as a therapeutic procedure prior to surgery or as an alternative. Surgical neuroangiography is now also a well-established therapeutic technique in neuropediatrics. Lesions fed by the external carotid artery and spinal cord lesions are the main indications. The first group consists of maxillofacial vascular malformations, nasopharyngeal angiofibromas, and vascularized tumors of the facioorbital area (hemangiopericytoma, angiosarcoma ...). Spinal lesions for which embolization can be considered are benign tumors of the vertebral column (vertebral hemangioma, aneurysmal bone cyst ...) and vascular malformations intrinsic to the spinal cord. Particles (Ivalon, dura) are the safest embolization materials and have a wide range of possible applications; glues such as IBC have defined but limited indications. Digital subtraction angiography is crucial during the procedure and in determining the overall doses of contrast medium to be administered. Furthermore, decisions can be made more rapidly and precisely, with a definite improvement in the therapeutic results. PMID- 2544267 TI - Synthesis of analogs of N-acetylneuraminic acid and their effect on CMP-sialate synthase. PMID- 2544270 TI - Acalculous cholecystitis complicating hepatic intraarterial lipiodol: case report. AB - Hepatic intraarterial injection of Lipiodol has been used by various authors to enhance the diagnostic accuracy of computed tomography in hepatic tumors. The technique appears safe and seems free from serious complications when used judiciously. We report a case in which injection of hepatic intraarterial Lipiodol precipitated acalculous cholecystitis requiring urgent laparotomy. PMID- 2544268 TI - Intraarterial digital subtraction angiography in detection of hepatocellular carcinoma. AB - To evaluate the usefulness of intraarterial digital subtraction angiography (IADSA) in hepatocellular carcinoma, IADSA was prospectively performed, combined with conventional film-screen angiography, in 40 patients in whom the tumor was histologically proven or highly suspected. Of the 16 main tumors less than 2 cm, IADSA detected 7 that were missed by film-screen angiography; 6 of these were less than 1 cm in size. With other diagnostic modalities including ultrasound, routine CT study, and CT after intraarterial Lipiodol injection, the detection rate of these tumors was low. IADSA's contribution is important in the detection of small hepatocellular carcinomas. PMID- 2544272 TI - [Chronic fatigue syndrome]. PMID- 2544269 TI - Lipiodol retention within hepatic cavernous hemangioma. AB - Intraarterial injection of Lipiodol has been recommended to differentiate hepatocellular carcinoma from benign lesions such as cavernous hemangioma, because uptake and prolonged retention of the contrast medium is a characteristic of the malignant tumors. In two cases of cavernous hemangioma of the liver in which we injected Lipiodol, uptake and retention up to 3 months was demonstrated. We conclude that the intraarterial injection of Lipiodol may not be reliable in differentiating hepatocellular carcinoma from cavernous hemangioma of the liver. PMID- 2544271 TI - Milestones in clinical pharmacology. Remember the Sippy regimen for peptic ulcer? PMID- 2544273 TI - [The significance of oxygen radicals in the pathogenesis of interstitial pulmonary processes]. AB - The author assessed the production of superoxide anion radical (O2-) and hydrogen peroxide (H2O2) by monocytes in blood and by alveolar macrophages in five healthy non-smokers and in four non-smokers with a bioptically confirmed cryptogenic fibrotizing alveolitis under basal conditions and after stimulation of the cells with 25 microM forbol myristate acetate. For assessment of the production of O2- and H2O2 he used the colorimetric method described by Pick and Keisari. Alveolar macrophages of patients with cryptogenic fibrotizing alveolitis produce increased amounts of O2- and H2O2 (33.4 +/- 6.7 and 36.2 +/- 7.6 nmol/10(6) cells/hour resp.), as compared with alveolar macrophages of healthy volunteers (5.8 +/- 2.3 and 6.23 +/- 2.7 nmol/10(6) cells/hour resp.). Blood monocytes of patients with cryptogenic fibrotizing alveolitis do not have an increased production of these radicals, as compared with healthy non-smokers (3.97 +/- 1.4 and 4.26 +/- 1.7 resp., as compared with 3.79 +/- 1.2 and 3.87 +/- 1.1 nmol/10(6) cells/hour resp.). After stimulation with forbol myristate acetate alveolar macrophages in patients with cryptogenic fibrotizing alveolitis increase the output of oxide radicals only slightly. PMID- 2544274 TI - The organization of prolactin-like-immunoreactive neurons in the rat central nervous system. Light- and electron-microscopic immunocytochemical studies. AB - The localization and distribution of prolactin-like-immunoreactive perikarya and nerve fibers in the rat central nervous system have been studied by a preembedding immunoperoxidase method using well-characterized specific immunsera to rat prolactin. Although the localization of labeled neuronal structures in a number of brain areas correlates with the data of previous immunocytochemical studies, we found prolactin-immunoreactive neurons in various regions not previously reported. In untreated animals, the highest concentrations of prolactin-fibers were observed: (i) in the external layers of the median eminence where they exhibited close contact with blood vessels, and (ii) in the bed nucleus of the stria terminalis and in the central nucleus of the amygdala where they closely surrounded unlabeled perikarya. Dense networks of finely varicose prolactin fibers were also observed in the organum vasculosum of the lamina terminalis, in the subfornical organ, and in the dorsolateral regions of the medulla oblongata and the spinal cord. Lastly, a number of large, varicose, intensely immunoreactive fibers were found in the olfactory bulb, the cingulum, and the periventricular regions of the hypothalamus and central gray, whereas isolated fibers could be detected in the caudate nucleus and in the cerebral cortex. In animals treated with colchicine, prolactin-immunoreactive perikarya were essentially located within the periventricular and perifornical regions of the hypothalamus, and within the bed nucleus of the stria terminalis. Although corticotropin (ACTH 17-39)-immunoreactive fibers could be detected in several regions found to contain prolactin fibers, the distribution and organization of both fiber types clearly differed in numerous brain regions, and the regions containing the corresponding perikarya did not overlap.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544276 TI - [Serological survey of cytomegalovirus infection in Guizhou]. AB - From August to October 1987, 577 serum samples were examined for the presence of CMV-IgG antibody by using indirect ELISA in Guizhou. The positive rate of CMV antibody varied from 87.5 to 92.4% in three areas and from 80.9-94.9% in various ages groups. It was suggested that the main transmission route of CMV infection in early life might be transmitted from mothers to their infants, but there might be some close contact transmission in population. The results showed that CMV infection was common in Guizhou. It was necessary to study further the correlation between CMV infection and birth defects. PMID- 2544277 TI - [A survey of antibodies to arboviruses in residents in south-western part of Yunnan province]. AB - In order to provide serological evidence for arbovirus infection in Yunnan province, 760 Samples of human sera collected from nine counties in south-western part of Yunnan province were examined for HI antibodies to 11 arboviral antigens. The viruses used in this experiment included 3 alphaviruses (MAY, VEE and CHIK) and 8 flaviviruses (JE, MVE, KUN, DEN3, DEN4, KFD, LGT and POW). 275 samples were found positive for HI antibodies to alphaviruses (36.2%) and 189 of them (68.7%) reacted with MAY virus, 61 (22.2%) with CHIK. The GMT of HI titers for VEE, MAY and Chik were 164.4, 94.5 and 66.7, respectively. 588 samples of sera (77.4%) were found positive for HI antibodies to flaviviruses. The positive HI antibody rates were as follows: JE, 27.9%; DEN, 36.6%; KFD, 22.2%; MVE, 22.1%; KUN, 18.7%; POW, 8.8% and LGT, 7.9%, respectively. Its average GMT was 356, and the antibody titers of 403 samples of sera were higher than 1/640. Cross reactions among viruses, especially flaviviruses, were usually found by HI test, and superinfections were present. In addition to the existence of JE and DEN viruses the results clearly showed that many Kinds of arboviruses might exist in Yunan Province. PMID- 2544275 TI - Intracellular signals trigger ultrastructural events characteristic of meiotic maturation in oocytes of Xenopus laevis. AB - Oocytes of Xenopus laevis were treated with agents which induce individual intracellular signals normally evoked during the process of meiotic maturation. Ultrastructural analysis of these oocytes allowed identification of specific second messengers that individually trigger single ultrastructural changes characteristic of the meiotic maturation process: Manipulation of intracellular cAMP levels induced changes in cortical granule position. Cytoplasmic alkalinization triggered a disruption of the annulate lamellae, a specialized organelle in the periphery of oocytes. Activation of protein kinase C caused rapid formation of a cortical endoplasmic reticulum and subsequent disruption of cortical granules. Manipulation of transmembrane calcium flux had varied results dependent upon the agent employed. Two of the treatments, Verapamil and zero external calcium, induced a reorganization in the oocyte periphery. The results indicate that these ultrastructural events are under the control of specific intracellular signals known to be elicited during meiotic maturation. PMID- 2544278 TI - [Detection of specific antibodies to cytomegalovirus in the blood donors in Qingdao]. AB - The detection of cytomegalovirus (CMV) infection in the blood donors in China has not been reported the literature available at hands. 344 serum samples from healthy blood donors in Qingdao were examined for the presence of CMV-IgG by IFA and CMV-IgM, CMV-IgA by ELISA. The frequency of positive donors was 99,45%, 4,65%, and 3,20%, respectively. The result showed CMV infection was rather common in the donors, the rate of active CMV infection was also higher than that abroad. We suggest that when blood is transfused to high risk patients, CMV-IgM and CMV IgA should be examined regularly on donors to exclude or reduce the incidence of transfusion-associated CMV infection. At present it is not practical for CMV-IgG as a screening marker in our country. PMID- 2544279 TI - [A study on the relationship between reduced activity of red cell membrane sodium potassium pump and essential hypertension]. AB - A population-based matched study of 103 pairs on the relation of sodium-potassium pump defect and hypertension was conducted at a factory in 1986. Conditional Logistic regression technique was used in data analysis. It was suggested that the increase of RBCNa+ and decrease of RBCK+ and RBCNa+, K+ATP-ase were significantly associated with hypertension. Plasma sodium and body weight, which were considered as important risk factors of hypertension, were fund correlated with the decrease of RBCNa+, K+ATPase, it was proposed that reduced activity of cell membrane sodium-potassium pump might be the common link of the two factors with hypertension. PMID- 2544280 TI - The pharmacology of the novel 5-HT1-like receptor agonist, GR43175. AB - We describe the identification of a novel drug, GR43175, for the acute treatment of migraine. GR43175 is a tryptamine analogue with a very selective agonist action at a 5-HT1-like receptor subtype first identified in the dog saphenous vein. Using this drug as a research probe, we have now shown that this 5-HT receptor type predominates in the carotid circulation, which explains the remarkably selective vasoconstrictor action of GR43175 in vivo in the carotid arterial bed of dogs and cats. Its vasoconstrictor action can be shown to be localized even further to arteriovenous anastomoses (shunts) within the carotid circulation, in such a way that blood flow to the brain as well as to extracerebral capillary beds remains unaffected or may even be increased. In the treatment of migraine demonstrated to date, the impressive effectiveness of GR43175 must reinforce the evidence in favour of an important vascular component being involved in the aetiology of the disease. The question is again raised as to whether the opening of carotid shunts is involved, as suggested by Heyck. If not, an alternative vascular locus needs to be identified. PMID- 2544281 TI - GR43175 does not share the complex pharmacology of the ergots. AB - The aim of this study was to compare the effects of the selective 5-HT1-like receptor agonist, GR43175, with several ergot derivatives in several different pharmacological preparations. In the rat isolated uterus, ergotamine, dihydroergotamine, ergometrine and methylergometrine (all at concentrations of 1 microM) caused marked uterotonic activity whilst GR43175 (10 and 30 microM) was inactive. Similarly, in the pithed rat the ergot derivatives caused marked increases in diastolic blood pressure (in doses up to 1000 micrograms/kg intravenously), whilst GR43175 (in doses up to 1000 micrograms/kg intravenously) was inactive. In anaesthetized dogs, both ergotamine (0.3-1000 micrograms/kg intravenously) and GR43175 (1-1000 micrograms/kg intravenously) caused dose dependent vasoconstriction in the carotid artery bed. However, the effects of ergotamine, unlike those of GR43175, were accompanied by increases in diastolic blood pressure and coronary vasoconstriction. These studies provide further evidence for the remarkably selective vasoconstrictor action of GR43175. PMID- 2544282 TI - The selective closure of feline carotid arteriovenous anastomoses (AVAs) by GR43175. AB - The haemodynamic effects of the selective 5-HT1-like agonist GR43175 have been compared with that of ergotamine in anaesthetized cats. Both GR43175 (30-1000 micrograms/kg intravenously) and ergotamine (0.3-30 micrograms/kg intravenously) caused a dose-dependent reduction in the proportion of cardiac output passing through arteriovenous anastomoses (AVAs). However, unlike GR43175, the effect of ergotamine (30 micrograms/kg intravenously) was associated with marked increases in diastolic blood pressure and total peripheral resistance. In further studies, the effect of GR43175 on the distribution of blood flow within the carotid bed has been examined. GR43175 caused a reduction in total carotid arterial blood flow which was entirely due to a reduction in flow through carotid AVAs. These results demonstrate that GR43175, unlike ergotamine, has a highly selective vasoconstrictor action on AVAs within the cranial circulation of anaesthetized cats. Such a mechanism may be important in its antimigraine activity. PMID- 2544283 TI - Characterization of the 5-hydroxytryptamine receptor which mediates contraction of the human isolated basilar artery. AB - This paper reports part of a study which investigated the identity of the receptor involved in 5-hydroxytryptamine (5-HT) mediated contraction of the human basilar artery in vitro. 5-HT and a variety of 5-HT receptor agonists contracted human isolated basilar artery with a rank order of agonist potency: 5 carboxamidotryptamine greater than 5-HT greater than GR43175 much much greater than 2-methyl-5-HT. The maximum response produced by these agonists differed. The contractile responses to both 5-HT and GR43175 were resistant to antagonism by the 5-HT2 antagonist ketanserin and the 5-HT3 antagonist GR38032, indicating that neither 5-HT nor GR43175 activate 5-HT2 and 5-HT3 receptors in this tissue. In striking contrast, methiothepin (an antagonist at 5-HT1-like receptors) proved a potent antagonist of the contractile actions of both 5-HT and GR43175. Methiothepin did not antagonize the contractile response to the thromboxane-A2 mimetic U-46619. It is concluded that 5-HT and GR43175 contract the human isolated basilar artery by activating 5-HT1-like receptors. It is suggested that the antimigraine action of GR43175 may reflect its ability to constrict certain cranial arteries via 5-HT1-like receptor activation. PMID- 2544284 TI - The kinetics of 14C-GR43175 in rat and dog. AB - GR43175 is a selective 5-HT 1-like receptor agonist which is effective in the acute treatment of migraine. Rats and dogs were dosed intravenously (iv) and orally (po) with 1 mg 14C-GR43175 base (as succinate salt)/kg bodyweight. GR43175 is rapidly absorbed after oral dosing. In dog, 95-100% of the dose is absorbed, but less (25-30%) is absorbed by the rat. The bioavailability is greater than 54% in dog, but lower in rat. Except for the CNS, drug-related material is widely distributed after iv dosing, but is mainly concentrated in the gastrointestinal tract and excretory organs after oral dosing. GR43175 is eliminated from plasma by a combination of renal and metabolic clearance. Some first-pass metabolism occurs in both species. In dog the major route of excretion is in urine (78-83% of the dose) after either route of administration. In rat, urine is also the major route of excretion (71%) after iv dosing. After oral dosing to the rat the major route of excretion is in the faeces (83%). GR43175 is extensively metabolized, after either route of administration, in both species. GR49336, the indole acetic acid derivative of GR43175, is the major metabolite in dog and a major metabolite in rat. PMID- 2544285 TI - Early studies with the novel 5-HT 1-like agonist GR43175 in healthy volunteers. AB - A series of single dose studies has been undertaken after intravenous, subcutaneous and oral administration to healthy volunteers to determine safety, tolerability and pharmacokinetics of GR43175, a novel 5-HT 1-like receptor agonist. GR43175 proved to be well tolerated using all routes of administration. Minor side effects were reported including scalp tingling, sensations of bodily warmth, lightheadedness and heavyheadedness. In addition, short-lasting vasopressor effects were encountered, although no drug-related changes were seen in heart rate and ECG or in laboratory safety screens. The drug was rapidly absorbed after oral dosing and had a mean absolute bioavailability of 14% due at least in part to first-pass metabolism. Plasma concentrations in healthy volunteers fell within the range for producing relief of headaches in migraine patients. GR43175 had a high mean plasma clearance (1313 ml/min), the majority of which was due to non-renal clearance and a large volume of distribution (2351). The mean half-lives were 1.7 h and 2.3 h after intravenous and oral doses, respectively. PMID- 2544286 TI - Overview of initial clinical studies with intravenous and oral GR43175 in acute migraine. AB - The novel 5-HT 1-like receptor agonist GR43175 has been evaluated as a treatment for acute migraine in a series of open, dose-ranging and controlled clinical trials. Patients with severe attacks of migraine have attended special pain or headache clinics for treatment and assessment. Given intravenously as a bolus, GR43175 is capable of aborting all migraine symptoms within 10-30 min in over 90% of cases at a dose of 64 micrograms/kg. Characteristic transient and reversible side effects with such a regimen include feelings of heaviness, pressure and occasionally warmth or tingling which can be diminished by extending the duration of drug administration to a short infusion. Initial dose-ranging studies with a dispersible tablet formulation of GR43175 have revealed an efficacy of 70-85% within 2 h with doses of 70-280 mg. Furthermore, tolerability is excellent. These encouraging early results warrant larger-scale controlled studies of GR43175 in acute migraine. PMID- 2544287 TI - Early clinical experience with subcutaneous GR43175 in acute migraine: an overview. AB - In six European clinics 111 migraine patients were treated in a series of open dose-ranging studies with subcutaneous injections of 1 to 4 mg GR43175, a novel 5 HT 1-like receptor agonist. Response rates after 20-30 min were dose related and rose from 33% with 1 mg to 96% with 4 mg GR43175. Side effects were minor and transient. These promising results warrant confirmation in placebo-controlled double-blind trials. Migraine, treatment, 5-HT agonist, open studies. PMID- 2544288 TI - Initial clinical experience with the use of subcutaneous GR43175 in treating acute migraine. AB - Subcutaneous GR43175 was examined in patients with acute migraine for efficacy, tolerability and safety in an open, controlled, dose-ranging study. Ten patients with acute, non-medicated, migraine (15 attacks) were assessed for severity of headache and associated symptoms (nausea, vomiting and photophobia). GR43175 plasma samples were monitored serially after dosing. Doses of 2 mg or 3 mg gave rapid relief of all migraine symptoms. Thirteen attacks (86%) had either resolved completely or improved to a mild non-migraine residual headache within 40 min. Treatment was well tolerated at all doses, the only adverse effects being transient pain on injection. Peak plasma concentrations were obtained within 10 20 min; a decline in plasma drug concentration did not result in a relapse in headache severity. PMID- 2544289 TI - Treatment of acute migraine with subcutaneous GR43175 in West Germany. AB - A subcutaneous preparation of GR43175, a novel antimigraine 5-HT 1-like agonist, was considered to represent a convenient way of administering the drug to patients during an acute migraine attack. In a series of open, uncontrolled dose ranging studies, 82 patients with migraine were assessed serially for changes in severity of headache and associated symptoms following subcutaneous GR43175 in doses of 1-4 mg. Subcutaneous injection of 3 mg or 4 mg was found to be most effective. Within 60 min, 90% of patients had obtained complete relief of all migraine symptoms. Tolerability was good, 59% of patients reporting no adverse effects. Those reported mainly comprised transient local irritation to the injection. There were no changes attributable to GR43175 in heart rate, blood pressure, ECG readings or laboratory parameters. PMID- 2544290 TI - Effective improvement of symptoms in patients with acute migraine by GR43175 administered in dispersible tablets. AB - GR43175, a selective 5-HT 1-like agonist, was administered as oral dispersible tablets in an open, uncontrolled dose-ranging study to assess its efficacy as an agent for acute migraine. Nine patients, all with well established attacks, were assessed for changes in severity of headache and associated symptoms over 2 h. Drug absorption was compared during and between attacks in five patients. Doses of 140 mg and 280 mg resulted in complete relief of all symptoms within 2 h. Treatment was well tolerated in all patients. PMID- 2544291 TI - Lack of effect of GR43175 on peripheral arteries in man. AB - In a double-blind, placebo-controlled crossover study in eight male volunteers, toe-arm systolic gradients, measured with strain-gauge plethysmography, were measured for 4 h after 2 mg intravenous GR43175 or placebo. No changes were observed, indicating that GR43175 is without significant vasoconstrictor effect on peripheral arteries in man. PMID- 2544292 TI - Involvement of a type 1 protein phosphatase encoded by bws1+ in fission yeast mitotic control. AB - Fission yeast cdc25+ and wee1+ interact genetically with cdc2+ in the regulation of cell division, respectively as a mitotic activator and inhibitor. cdc25+ is normally essential for mitosis, but this requirement is alleviated in a loss-of function wee1 mutant background. A plasmid-borne sequence, other than wee1+, that causes a cdc25ts wee1- double mutant to revert to a temperature-sensitive cdc phenotype has been isolated. The gene carried by this plasmid is called bws1+ (for bypass of wee suppression). bws1+ also bypasses the ability of alleles of cdc2 that confer a wee phenotype (cdc2w) to suppress loss-of-function cdc25 mutants. The nucleotide sequence of bws1+ shows that the predicted protein shares 81% amino acid identity with the catalytic subunit of mammalian type 1 protein phosphatase. Thus a genetic screen that might have yielded a protein kinase (wee1+) uncovered a phosphatase that also appears to be involved in the pathway of mitotic control. PMID- 2544293 TI - Putting it on and taking it off: phosphoprotein phosphatase involvement in cell cycle regulation. PMID- 2544294 TI - Mapping replication units in animal cells. AB - A general approach for assaying the in vivo direction of replication for any DNA segment has been developed. This technique allows the scanning of genomic regions to detect bidirectional tail-to-tail replication, indicating the presence of a functional origin. By this criterion we identified the approximate positions of two origin sites downstream of the Chinese hamster DHFR gene. Further mapping revealed areas of head-to-head replication, signifying locations of replication termination and thus defining the landmarks of a complete animal cell replicon. Genetic proof for the existence of the DHFR origin was obtained by showing that this region serves as a bidirectional DNA synthesis initiation point following its integration into other sites in the genome by transfection. To show the general applicability of this methodology, we studied the APRT domain. Replication mapping together with the use of deletion mutants allowed the identification of an origin at a far-upstream locus. PMID- 2544295 TI - A recombination hotspot in the LTR of a mouse retrotransposon identified in an in vitro system. AB - The recombinational frequency between two long terminal repeat elements (LTR-IS) of a mouse retrotransposon was about 13 times higher, compared with that of two control DNA sequences in extracts from mouse testes, but not in extracts from ascites cells. Deletion of a 37 bp region from the LTR-IS element strongly suppresses its recombinational activity. This 37 bp region encompasses an area of potentially single-stranded DNA and interacts with at least two nuclear proteins. One of them binds sequence-specifically to single-stranded DNA and is present in both types of extracts. Another protein(s) binds to dsDNA at the motif TGGAAATCCCC and is absent in extracts from testes. Our results suggest that a cis acting DNA sequence within the 504 bp LTR-IS element is responsible for its high recombinational activity in vitro, and they further support the previous suggestion that the LTR-IS elements are meiotic recombinational hotspots in vivo. PMID- 2544296 TI - A subthreshold level of DNA topoisomerases leads to the excision of yeast rDNA as extrachromosomal rings. AB - In a yeast DNA topoisomerase double mutant TG205 (delta top1 top2-4), over half of the rDNA is present as extrachromosomal rings containing one 9 kb unit of the rDNA gene or tandem repeats of it. Expression of a plasmid-borne TOP1 or TOP2 gene in the strain leads to the integation of the extrachromosomal rDNA rings back into the chromosomal rDNA cluster. When the plasmid-borne topoisomerase gene is expressed from an inducible promoter of the GAL1 gene, repression of the gene by dextrose leads to reappearance of the extrachromosomal rDNA rings. The DNA topoisomerase-dependent excision/integration of rDNA is discussed in terms of the possibility of rDNA supercoiling by transcription and the effects of DNA topology on intra- and interchromosomal recombination. PMID- 2544297 TI - The bimG gene of Aspergillus nidulans, required for completion of anaphase, encodes a homolog of mammalian phosphoprotein phosphatase 1. AB - In Aspergillus nidulans, the temperature-sensitive, recessive cell cycle mutation bimG11 causes an elevated mitotic index at restrictive temperature and an inability to complete the anaphase separation of daughter nuclei. We have shown that this mutation has an abnormally high content of nuclear phosphoproteins and that the wild-type gene encodes a type 1 protein phosphatase. We conclude that dephosphorylation of a key protein(s) is required to complete mitosis. PMID- 2544299 TI - Protein disulfide isomerase: multiple roles in the modification of nascent secretory proteins. PMID- 2544298 TI - The fission yeast dis2+ gene required for chromosome disjoining encodes one of two putative type 1 protein phosphatases. AB - S. pombe dis mutants block mitotic chromosome disjunction in a manner reminiscent of aneuploidy formation, and belong to three distinct genes, dis1-dis3. We cloned two independent genomic DNAs that complemented both the cold-sensitive and caffeine-hypersensitive phenotype of dis2-11. These genes, dis2+ and a suppressor sds21+, encode proteins (calculated MW 37,000) with similar predicted amino acid sequences. dis2+ and sds21+ have overlapping functions, and disruptants are lethal only when both genes are disrupted. The gene products identified by anti dis2 serum are enriched in nuclei. By hybridization, we obtained two cDNA clones from mouse and one genomic clone from S. cerevisiae; the latter complements S. pombe dis2-11. These dis2+ and similar polypeptides of yeasts and mouse are found to be highly homologous (75%-90% identical) to rabbit protein phosphatase 1. The implications of these findings are discussed with regard to mitotic control. PMID- 2544300 TI - Activation of neutrophils by recombinant interleukin 6. AB - The cytokine interleukin 6 (IL-6) has been shown to have multiple biological activities against many cellular targets. The present studies were designed to determine whether these activities extended to the neutrophil (PMN). Initially, we investigated the ability of IL-6 to modulate PMN-mediated antibody-dependent cellular cytotoxicity. The presence of IL-6 stimulated 51Cr release from labeled, opsonized targets by 67.1% (from 21.6 +/- 1.4% to 36.1 +/- 1.3% at 10 U of IL-6 (P less than 0.01)). IL-6 was not directly toxic to the target cells and stimulation of ADCC was shown to occur across a range of effector-to-target ratios. To investigate the basis of the capacity of IL-6 to stimulate PMN, we studied the effects of IL-6 on PMN chemotaxis, degranulation, and the respiratory burst. IL-6 was not chemotactic or chemokinetic for PMN. However, IL-6 stimulated lysozyme secretion from 14.1 +/- 2.5 to 23.7 +/- 3.6% at 100 U (P less than 0.01). IL-6 was a complete secretagogue, being able to induce the secretion of both the secretory granule marker lactoferrin (11.2 +/- 2.0 to 23.5 +/- 2.2%) and the primary granule marker beta-glucuronidase (5.0 +/- 1.0 to 18.2 +/- 4.0%). IL 6 was not able to directly stimulate the PMN respiratory burst. However, IL-6 did "prime" PMN, enhancing superoxide secretion by fMLP (10(-7) M)-treated PMN by 50.8% (5.9 +/- 1.0 to 8.9 +/- 1.5 nmol superoxide at 100 U of IL-6; P less than 0.01) and PMA (5.0 nM) by 54.3% (8.1 +/- 2.6 to 12.5 +/- 3.6 nmol; P less than 0.05). In conclusion, IL-6 is a PMN stimulant, enhancing the toxicity of PMN in an antibody-dependent cellular cytotoxicity assay. Enhanced cytotoxicity may have been mediated, at least in part, by the stimulation of secretion of toxic components from PMN targets and by the priming of stimulating respiratory burst activity. PMID- 2544301 TI - Wild poliovirus isolation in the Americas, 1988. Pan American Health Organization. PMID- 2544302 TI - Two institutional outbreaks of Norwalk-like gastroenteritis--Ontario. PMID- 2544303 TI - [BCG-polysaccharides-nucleic acid and its effect in the prophylactic treatment of the common cold]. AB - The polysaccharides-nucleic acid fraction extracted from BCG was found out to be a good adjuvant. Through the activation and protective of the suppressed body immune system, particularly the macrophages and T cells, it gave good protection to the Sendai virus airway infection of C57BL mice, and clinical effect on the common cold susceptible. PMID- 2544304 TI - [Combined modality treatment of small cell lung cancer by chemotherapy, radiotherapy, immunotherapy and Chinese traditional medicine]. AB - The combined modality treatment consisted mainly of long-term intermittent combination chemotherapy (VCMB being in the majority) and radiotherapy combined with immunotherapy as well as Chinese traditional medicine. The patients treated with this therapy were observed in 1974-1986, till successive fifty, and followed up to January 1988. They were of the stage I, II, III and IV including 3, 10, 20 and 17 patients respectively. The complete remission rate was 54% (27/50); the partial remission rate, 40% (20/50). The total effective rate was 94% (47/50). According to the calculation of life-table, 2-year survival rate for the stage I II (n = 13) was 76.2%; 5-year survival rate, 55.5%. For the stage III (n = 20), 2 year survival rate was 46.9%; 5-year, 18.8%. By long-term combined modality, the survival rate has obviously improved, and the possibility of cure has evidently increased. PMID- 2544305 TI - Synthesis and purification of 5,5-dimethyl-1-pyrroline-N-oxide for biological applications. AB - General procedures for the synthesis and purification of 5,5-dimethyl-1-pyrroline N-oxide (DMPO) and its synthetic intermediates for electron paramagnetic resonance (EPR) spin trapping is described. The synthetic methods are not new per se and are based on Todd's original scheme (1959). In contrast to his account, however, we provide a detailed description of the numerous procedures and precautions necessary to obtain DMPO of very high 'chemical' and (most importantly) 'EPR' purity for use in biological spin trapping. PMID- 2544306 TI - Determination of subcutaneous tumor size in athymic (nude) mice. AB - The athymic (nude) mouse is a useful model for studying the biology and response to therapies of human tumors in vivo. A survey of recent literature revealed the use of 19 different formulas for determining the size of subcutaneous tumors grown as xenografts in nude mice (2 for determining tumor area, 3 for tumor diameter, and 14 for calculating tumor volume). We compared the volumes, areas, and diameters predicted by each of the 19 formulas with the actual weights of 50 tumors ranging from 0.46 to 22.0 g established in nude mice as xenografts from human cell lines. In addition to determining how well each formula predicted relative tumor size, we analyzed how well each formula estimated actual tumor mass. The ellipsoid volume formulas (pi/6 x L x W x H and 1/2 x L x W x H) were best for estimating tumor mass (r = 0.93), whereas measurements of diameter correlated poorly with tumor mass (r less than 0.66). Although determination of tumor area correlated well with tumor mass in small tumors (r = 0.89), correlations of area with tumor mass for large tumors were poor (r = 0.41). We conclude that determination of the ellipsoid volume from measurements of three axes consistently yields the most accurate estimations of both relative and actual tumor mass. PMID- 2544307 TI - Inhibition of topoisomerases by fredericamycin A. AB - Fredericamycin is an antibiotic product of Streptomyces griseus that exhibits modest antitumor activity in vivo and in vitro. Because of its unique structure and the absence of a clearly defined mechanism of action, we examined the effects of this compound on L1210 cells in culture as well as on several enzymes that bind to DNA. Fredericamycin exhibits an IC50 of 4.4 microM toward L1210 cells, and its cytotoxicity is a function of the time of exposure as well as drug dose. No DNA breakage was observed in L1210 cells or isolated nuclei following exposure to highly lethal concentrations of fredericamycin. As a first step toward understanding its mechanism of action, we examined the effect of fredericamycin on several enzymes involved in DNA metabolism. The catalytic activity of both DNA topoisomerases I and II were totally inhibited by fredericamycin concentrations of 4.4 and 7.4 microM, respectively. Fredericamycin blocked etoposide-stimulated DNA cleavage by topoisomerase II both in vitro and in isolated nuclei. In addition, the drug inhibits DNA polymerase a in vitro, exhibiting an IC50 of 93 microM. These diverse actions of fredericamycin do not enable us to draw conclusions regarding its mechanism of antitumor effect but clearly identify it as a compound of pharmacologic interest. PMID- 2544308 TI - Responsiveness of human lung cancer/nude mouse to antitumor agents in a model using clinically equivalent doses. AB - The responses of 14 lines of human lung cancer xenografts in BALB/c-nu/nu mice to eight known antitumor agents were investigated. These xenografts consisted of four small-cell carcinomas (SCLC) and ten non-small-cell carcinomas (four large cell, three squamous cell, and three adenocarcinomas; NSCLC). The doses used in the experiments were the maximum tolerated dose (MTD) in nude mice and the "rational dose" (RD), the latter considered to be pharmacokinetically equivalent to the clinical dose. When given at MTDs, all drugs except 5-fluorouracil (5-FU) and methotrexate (MTX) were extremely effective against NSCLC as well as SCLC. The response rates of drug-sensitive SCLC to mitomycin C (MMC), ACNU, and vinblastine (VLB) were 100%, and those to Adriamycin (ADR) and vincristine (VCR) were 75%. In addition, the response rates of even drug-resistant NSCLC to MMC and VLB were 70% and 90%, respectively. In contrast, the response rates of NSCLC to RDs of the drugs were reduced to less than 40% and corresponded well to the respective clinical rates. In SCLC, a good correlation of experimental and clinical response rates was observed with four drugs [cyclophosphamide (CPM), ACNU, VLB, and 5-FU]. As a result, we emphasize that a more reasonable prediction of the clinical effectiveness of antitumor agents can be made by a protocol using clinically equivalent doses. PMID- 2544309 TI - A phase I study of a new 5HT3-receptor antagonist, BRL43694A, an agent for the prevention of chemotherapy-induced nausea and vomiting. AB - In a phase I study of BRL43694A, a 5HT3-receptor antagonist, a single dose of 40 micrograms/kg was given to 24 patients. All patients received cytostatic treatment expected to cause nausea and vomiting. During the first 24 h, 12 patients were completely protected from nausea and vomiting, 4 experienced nausea and 8 had moderate vomiting; mild headache occurred in 10 patients. No cardiovascular (including ECG) changes took place. Apart from headache, no neurological side effects occurred. PMID- 2544311 TI - Repair of 8-methoxypsoralen + UVA-induced damage in specific sequences in chromosomal and episomal DNA in human cells. AB - A study of the repair of DNA damage in the dihydrofolate reductase (dhfr) gene of SV40-transformed human fibroblasts after treatment with 8-methoxypsoralen (8MOP) and UVA is described. 8MOP+UVA-induced cross-links in the dhfr gene were completely repaired by 12 h in one normal and one Fanconi's anaemia (FA) group A cell line. In contrast, approximately 35% of cross-links in an episomally maintained Epstein--Barr virus derived plasmid remained unrepaired even after 48 h. Cross-linkable monoadducts in the dhfr gene were repaired more slowly than cross-links, and there was no detectable repair of cross-linkable monoadducts in the plasmid. Thus the ability of a cell to repair 8MOP+UVA-induced cross-links or cross-linkable monoadducts in an episome does not reflect its capacity to repair such lesions in genomic DNA. PMID- 2544310 TI - A new sensitive 32P-postlabeling assay based on the specific enzymatic conversion of bulky DNA lesions to radiolabeled dinucleotides and nucleoside 5' monophosphates. AB - A new sensitive 32P-postlabeling assay for DNA adducts has been developed in which DNA is hydrolyzed initially by nuclease P1 and prostatic acid phosphatase instead of micrococcal nuclease and spleen phosphodiesterase as employed in previous postlabeling procedures. When DNA containing bulky adducts, X1, X2, .....Xn, is digested with nuclease P1 at pH 5, normal nucleotides are released as 5'-monophosphates, pN, while adducts are excised as 5'-phosphorylated dinucleotides, pXipN, because internucleotide linkages on the 3' side of X resist attack by nuclease P1. Addition of prostatic acid phosphatase to such a digest results in 5'-dephosphorylation of the nucleotides to normal nucleosides, N, and adducted dinucleotides, XipN, carrying a 5'-terminal free hydroxyl group. The dinucleotides but not nucleosides are converted to 5'-32P-labeled dinucleotides, [32P]pXipN, by T4 polynucleotide kinase-catalyzed [32P]phosphate transfer from [gamma-32P]ATP. Upon mapping on polyethyleneimine--cellulose anion-exchange TLC, the labeled dinucleotide adducts produce characteristic autoradiographic fingerprints. Alternatively, they are further digested with snake venom phosphodiesterase to yield 5'-monophosphates, [32P]pXi and pN. TLC profiles of the monophosphate adducts are distinct from those of the dinucleotides. These reactions provide the basis of the new 32P-postlabeling scheme, which is compared in this paper with a previously reported protocol yielding adducts in the form of 5'-32P-labeled 3',5'-bisphosphates, [32P]pXip. The results show that the availability of three different types of 32P-postlabeled derivatives for the same adduct aids in the analysis and chromatographic characterization of DNA adducts from diverse exogenous and endogenous sources. PMID- 2544312 TI - Molecular mechanisms of alkylation sensitivity in Indian muntjac cell lines. AB - The responses of two Indian muntjac cell lines to two monofunctional alkylating agents were investigated. An SV40-transformed line (SVM) had an increased sensitivity to cell killing when compared to the other, euploid line (DM) after exposure both to methyl nitrosourea (MNU) and to dimethylsulphate (DMS) and also exhibited higher frequencies of sister chromatid exchanges (SCEs) following alkylation. The hypersensitivity of SVM to DMS correlates with the defective repair of single-strand breaks that results in the generation of long-lived breaks in the DNA following exposure, leading eventually to the formation of chromosome aberrations. In contrast no difference is seen in the formation of long-lived breaks in the DNA of SVM and DM after treatment with biologically relevant doses of MNU; in this case hypersensitivity may be due to the loss of O6 alkylguanine-DNA-alkyltransferase activity. The conclusion that the hypersensitivites of SVM to MNU and to DMS have different molecular bases is supported by transfection of SVM with plasmids containing the protein coding region of the Escherichia coli ada+ gene; subsequent expression within the cell corrects its hypersensitivity to the cytotoxic and SCE-inducing effects of MNU but has very little influence upon the lethality, SCE induction or the repair of long-lived DNA strand breaks after exposure to DMS. PMID- 2544313 TI - Differential inhibition by staurosporine, a potent protein kinase C inhibitor, of 12-O-tetradecanoylphorbol-13-acetate-caused skin tumor promotion, epidermal ornithine decarboxylase induction, hyperplasia and inflammation. AB - The effect of staurosporine on 7,12-dimethylbenz[a]anthracene (DMBA)-initiated and 12-O-tetradecanoylphorbol-13-acetate (TPA)-promoted skin papilloma formation was examined in CD-1 mice. A topical application of staurosporine 15 min prior to each TPA treatment resulted in a dose-related inhibition of tumor formation. Staurosporine by itself had no tumor producing activity in DMBA-initiated mice. Staurosporine failed to prevent TPA-induced edema formation, whereas quercetin markedly suppressed it. Staurosporine by itself did not induce a significant edema. Histological studies revealed that staurosporine failed to inhibit TPA induced inflammation but rather augmented TPA-induced polymorphonuclear leukocyte (PMN) infiltration. Staurosporine by itself induced a slight PMN infiltration 1 h after the drug application, but the effect was only transient. Although staurosporine failed to inhibit the TPA-induced epidermal hyperplasia and DNA synthesis significantly, nuclear atypism of the superficial layer of the epidermis appeared to be less remarkable in staurosporine-pretreated mice. TPA caused epidermal ornithine decarboxylase (ODC) induction was not inhibited by staurosporine but rather augmented by this agent. TPA enhanced the phosphorylation of 34 kd protein in intact epidermal cells in a concentration dependent manner. Staurosporine and 1-(5-isoquinolinylsulfonyl)-2 methylpiperazine (H-7) suppressed the TPA-stimulated phosphorylation of 34 kd protein, but palmitoylcarnitine failed to suppress it. In addition, TPA stimulated superoxide generation of rabbit peritoneal PMN was potently inhibited by staurosporine. It is possible that TPA induces inflammation, ODC activity, epidermal hyperplasia and tumor promotion through the activation of different type(s) of protein kinase C and staurosporine inhibits only certain type(s) of protein kinase C. Another possible explanation is that the protein kinase C inhibition by staurosporine depends on the nature of the substrate proteins or the intracellular localization of the enzyme. PMID- 2544314 TI - Monocyte-derived interleukin 1: effects on norepinephrine-stimulated aortic contraction and phosphoinositide turnover. AB - Medium conditioned by silica-stimulated human peripheral blood monocytes expresses vascular suppressive activity. Rat aortic rings, after incubation in conditioned medium, exhibited comprised contraction to stimulation by norepinephrine (NE). Maximal contraction (300 +/- 51 mg tension/mg tissue) and sensitivity (-5.91 +/- 0.23 M [log EC50]) were both reduced in comparison to contraction (762 +/- 66) and sensitivity (-7.42 +/- 0.11) displayed by rings after incubation in control medium. A polyvalent antibody (Ab) against human interleukin 1 (Il-1) neutralized the suppressive activity in conditioned medium. Rings incubated in conditioned medium containing Ab exhibited normal maximal contraction (722 +/- 46) and a partial restoration of sensitivity to NE (-6.91 +/ 0.13). In contrast, incubation of rings in control medium supplemented with recombinant human Il-1 resulted in a dose-dependent suppression of aortic contraction to NE that was analogous to the defects induced by monocyte conditioned medium. No significant differences in NE-stimulated phosphoinositide hydrolysis were present between rings incubated in Ab-treated or untreated conditioned or control media. The data suggest that monocyte-derived Il-1 may have a significant influence on vascular contractile function and that the mechanism by which Il-1 induces vascular dysfunction cannot be demonstrated to involve inhibition of NE-stimulated phosphoinositide metabolism. PMID- 2544315 TI - Equine peritoneal macrophage production of thromboxane and prostacyclin in response to platelet activating factor and its receptor antagonist SRI 63-441. AB - The formation of eicosanoids may be a primary route through which platelet activating factor (PAF) exerts its effects during endotoxemia. Since endotoxemia is a common cause of death in horses, a study was conducted to determine whether PAF could stimulate equine macrophage release of thromboxane A2 (TxA2) and prostacyclin (PGI2) and whether a PAF-receptor antagonist would alter macrophage eicosanoid synthesis. Equine peritoneal macrophages were cultured from clinically normal horses and exposed to various concentrations of PAF, the PAF-receptor antagonist SRI 63-441, endotoxin, or a combination of these. The supernatant concentrations of TxB2 and 6-keto-prostaglandin F1 alpha were determined after 6 hr incubation. The media concentration of TxB2 was increased significantly above baseline after treatment of macrophages with PAF (10(-7) to 10(-5) M), and the magnitude was similar to that induced by endotoxin. This TxB2 increase was not prevented by prior treatment of macrophages with SRI 63-441. SRI 63-441 (greater than or equal to 5 x 10(-5) M) significantly enhanced macrophage TxA2 synthesis, as well as its production of PGI2, similar to the effects of endotoxin. Media concentrations of 6-keto-prostaglandin F1 alpha were not increased significantly above baseline after treatment of macrophages with PAF (10(-8) to 10(-5) M). These results suggest that PAF may cause increased TxA2 release during endotoxemia, which may not be preventable by use of the PAF-receptor antagonist SRI-63-441, which is capable of inhibiting PAF-induced aggregation of equine platelets. PMID- 2544316 TI - Biological actions and properties of endothelium-derived nitric oxide formed and released from artery and vein. PMID- 2544317 TI - Opiate receptor antagonism in right-sided congestive heart failure. Naloxone exerts salutary hemodynamic effects through its action on the central nervous system. AB - Opiate receptor inhibition causes adrenergic receptor-mediated increases in aortic pressure, cardiac output, and left ventricular contractile function in right heart failure. To study whether the effects of opiate receptor inhibition are mediated by means of an action on the central opiate system, we administered equimolar doses of naloxone hydrochloride and naloxone methobromide (MeBr) and normal saline to heart failure dogs. Chronic stable right heart failure was produced by progressive pulmonary artery constriction and tricuspid valve avulsion. Naloxone hydrochloride caused an increase in mean aortic pressure, cardiac output, left ventricular dP/dt and dP/dt/P, plasma catecholamines, and regional blood flows to the myocardium, quadriceps muscle, kidneys, and splanchnic beds. Plasma beta-endorphin and adrenocorticotropin also increased. In contrast, neither normal saline nor naloxone MeBr (which does not cross the blood brain barrier) affected the systemic or regional hemodynamics or neurohormones. Naloxone hydrochloride was also administered to anesthetized heart failure dogs. Pentobarbital anesthesia removed cortical perception of nociceptive stimulation, reduced the increase in plasma epinephrine, and abolished vasodilation in skeletal muscle that occurred in conscious dogs after naloxone hydrochloride administration but had no major effects on responses of plasma norepinephrine, systemic hemodynamics, or other regional blood flows to opiate receptor inhibition. Naloxone hydrochloride had no effect in sham-operated dogs. The results indicate that the hemodynamic effects of naloxone are mediated by an action within the central nervous system. Furthermore, since pentobarbital anesthesia did not markedly alter the hemodynamic responses to naloxone hydrochloride, the acute salutary effects of opiate receptor inhibition probably are not caused by removal of the antinociceptive effect of endogenous opioids in heart failure. PMID- 2544318 TI - Characterization and pharmacological relevance of high affinity binding sites for [3H]LY186126, a cardiotonic phosphodiesterase inhibitor, in canine cardiac membranes. AB - [3H]LY186126, an analogue of the cardiotonic agent indolidan, was shown to bind reversibly and with high affinity (Kd = 4 nM) to a single class of binding sites within canine myocardial vesicles. Binding site density measured in various cardiac membrane fractions correlated well with Ca2+-ATPase activity (r = 0.94; p less than 0.01), but not with Na+,K+-ATPase or azide sensitive ATPase, indicating a localization of these sites within sarcoplasmic reticulum membranes. Divalent cations were required for binding and displayed the following order of activation: Zn2+ greater than Mn2+ greater than Mg2+ greater than Ca2+. Differential activation of [3H]LY186126 binding by various divalent cations was due to alterations in binding site density, rather than affinity. cGMP and selective inhibitors of type IV membrane-bound phosphodiesterase (SR-PDE), for example, indolidan, milrinone, imazodan, and enoximone, selectively displaced bound [3H]LY186126 caffeine, theophylline, and rolipram were relatively impotent as inhibitors of radiolabel binding. Kd values from displacement curves were highly correlated with IC50 values for inhibition of SR-PDE (r = 0.92; p less than 0.001). In addition, Kd values correlated well with published ED50 values for increases in cardiac contractility in pentobarbital-anesthetized dogs (r = 0.94; p less than 0.001). The results support the hypothesis that [3H]LY186126 labels the pharmacological receptor for the class of positive inotropic agents characterized as isozyme-selective phosphodiesterase inhibitors. Furthermore, the data suggest that the identity of the site labeled by [3H]LY186126 is SR-PDE, the type IV isozyme of cardiac phosphodiesterase located in the sarcoplasmic reticulum. PMID- 2544320 TI - Two components of calcium channel current in embryonic chick skeletal muscle cells developing in culture. AB - The properties of the Ca channel currents in chick skeletal muscle cells (myoballs) in culture were studied using a suction pipette technique which allows internal perfusion and voltage clamp. The Ca channel currents as carried by Ba ions were recorded, after suppression of currents through ordinary Na, K and Cl channels by absence of Na, K and Cl ions, by external TEA, by internal EGTA and by observing the Ba currents instead of the Ca currents. Two components of Ba current could be distinguished. One was present only if the myoballs were held at relatively negative holding potentials below -50 mV. This component first became detectable at clamp potentials of about -50 mV and reached a maximum between -10 and -20 mV. During long clamp steps, it became inactivated completely. The inactivation process of this component at a clamp potential of -30 mV was well fitted to a single exponential with a time constant of about -20 ms. Half-maximal steady-state inactivation was observed at -63 mV. The other component persisted even at relatively positive holding potentials above -40 mV, was observed during clamp pulses to -20 mV and above, and reached a maximum between +10 and +20 mV. This component inactivated very little; a substantial fraction of this component remained at the end of clamp pulses lasting 1 s. The inactivation process of this component at a clamp potential of -10 mV apparently followed a single exponential with a time constant of about 1 s. Half-maximal steady-state inactivation was attained at -33 mV. Both components of Ba current were blocked by Co ions, but organic Ca channel blocker D600 preferentially blocked the high-threshold, slowly inactivating component. The relationship between the current amplitude and the concentration of the external Ba ions was different between the two components. Furthermore, the two components of Ba current also differed in their developmental profile. These findings demonstrate the existence of two distinct types of Ca channels in the early stages of chick muscle cell development. PMID- 2544319 TI - Alterations in calcium antagonist receptors and sodium-calcium exchange in cardiomyopathic hamster tissues. AB - The Syrian cardiomyopathic (CM) hamster (BIO 14.6) develops a progressive cardiomyopathy characterized by cellular necrosis, hypertrophy, and, eventually, cardiac dilatation and congestive heart failure. Several lines of evidence implicate cellular calcium overload as an important etiologic factor. We previously reported an increased number of receptors for calcium antagonist drugs, which block voltage-dependent calcium channels, in heart, skeletal muscle, and brain tissue of these hamsters in the early necrotic stage of the disease. To better characterize the pathophysiological significance of this abnormality we evaluated calcium antagonist receptor binding and Na+-Ca2+ exchange in CM and control hamsters at different stages of disease as documented by quantitative histopathologic assessment. In CM hamsters as young as 10 days, an age previously thought to be before the onset of disease, we identified cardiac myocyte hypertrophy, a twofold increase in calcium antagonist receptor binding in heart and brain, and a 50% increase in skeletal muscle. Overt histological lesions were present in skeletal muscle at 25 days and in heart between 28-30 days. The size of cardiac lesions increased over time and changed from necrotic foci with cellular infiltration to fibrotic or calcified lesions by 360 days. Myocardial cellular hypertrophy persisted through the late stages of the disease (360 days), but increased calcium antagonist binding was present in heart only to 6 months of age, in skeletal muscle to 90 days, and in brain to 30 days. Na+-Ca2+ exchange in heart was normal until 15 days and then increased by 400% at 30 days suggesting that this augmentation might be a secondary response to the earlier increase in calcium antagonist receptors. At 360 days cardiac Na+-Ca2+ exchange was decreased by 50%, likely reflecting progressive cardiac damage. The increase in calcium antagonist receptors in CM animals as young as 10 days supports the hypothesis that abnormalities in voltage-dependent calcium channels play a role in the pathophysiology of CM hamsters. PMID- 2544321 TI - Divergent ontogeny of sigma and phencyclidine binding sites in the rat brain. AB - The postnatal developmental patterns of sigma (sigma) and phencyclidine (PCP) binding sites were compared in the rat brain. The results show diametrically different ontogenic patterns for the sites. While both the affinity and the density of sigma sites remain constant throughout the developmental period tested (postnatal day 1 to 1 year), the density of PCP binding sites increases from the time of birth, reaching the adult level by postnatal day 14. The differences in developmental patterns provide evidence for distinctive properties of cerebral sigma and PCP binding sites. PMID- 2544322 TI - Evidence for the involvement of nigral GABAB receptors in seizures of rat pups. AB - The substantia nigra GABA-sensitive system is important for the modification of seizures. In adult rats, nigral infusions of baclofen (GABAB agonist) had no effect on flurothyl seizures. This suggests that the nigral GABAB receptor system may not be involved in the mediation of flurothyl seizures in this age group. The present study examines whether the nigral GABAB receptor is involved in the modification of flurothyl seizures in rat pups and whether systemic infusions of baclofen could alter their seizure susceptibility. Baclofen (50-200 ng/0.25 microliters) was intranigrally administered to 16-day-old rat pups which were then exposed to flurothyl seizures. Results indicated that intranigral infusions of baclofen (100 and 200 ng/0.25 microliters) protected the rat pups against seizures. Systemic injections of baclofen also protected pups from flurothyl seizures suggesting that it may be worthwhile to examine baclofen as a potential antiepileptic drug in children. PMID- 2544323 TI - Intramuscular administration of iron during long-term chelation therapy with 2,3 dimercaptosuccinic acid in a man with severe lead poisoning. AB - 2,3-Dimercaptosuccinic acid (DMSA) an investigational chelant structurally similar to dimercaptopropanol (BAL), offers the advantage of not depleting iron stores on which basis it would not seem to form a toxic chelate with iron. We report the case of a man with a formidable body burden of lead (Pb) and depleted iron stores who was given iron intramuscularly during a defined period of long term retreatment with DMSA. Initiation of retreatment with DMSA, 30 mg/kg/day given orally in three divided doses for the first 7 days markedly enhanced Pb diuresis, entailed a pronounced fall in blood Pb and abolished symptoms of Pb poisoning. Continuation of retreatment with two-thirds the initial DMSA dose for an added 15 days maintained blood Pb at sustained low levels. Iron sorbitol administered intramuscularly during this period in individual doses of 100 mg of elemental iron given 3 days apart to a conservative total of 400 mg produced no untoward effects, suggesting that a toxic chelate between iron and DMSA was not formed. Serum ferritin entered the normal range and there was virtually an immediate significant decrease in erythrocyte protoporphyrin. Together with discernible increases in haemoglobin, haematocrit and MCV, this pointed to enhanced iron utilization. Since iron utilization is curtailed by high concentrations of Pb, the immediacy and magnitude of the post-chelation rebound in blood Pb precluded iron administration at any other stage. From these data, DMSA emerges as a uniquely versatile new chelant. Suitable for long-term administration, it permits the simultaneous parenteral administration of iron during dose-related sustained decreases in blood Pb. PMID- 2544324 TI - Correlations among blood levels of renin-angiotensin-aldosterone, bradykinin, and catecholamines in healthy subjects. PMID- 2544325 TI - Depression of humoral responses and phagocytic functions in vivo and in vitro by fish oil and eicosapentanoic acid. AB - Previous studies have demonstrated that eicosapentanoic acid (EPA) has anti inflammatory properties in both humans and experimental animals and may also depress humoral immunity in experimental animals. Our investigations showed that the addition of eicosapentanoic acid to human peripheral blood mononuclear cell cultures inhibited B cell responses to mitogenic stimulation and depressed the expression of interleukin 2 receptors in pokeweed mitogen-stimulated lymphocytes. Neutrophils were also affected in their ability to release the contents of primary and secondary granules, particularly when stimulated with antigen antibody complexes. Similar depressions of B cell responses and neutrophil functions were observed in a normal volunteer who ingested 6 g/day of a commercially available fish oil extract (equivalent to 2.1 g of EPA/day) during a 6-week period. Phagocytosis, enzymatic release, circulating immunoglobulin levels, and the response to tetanus toxoid both in vivo and in vitro were depressed during ingestion of fish oil. Most parameters showed a trend toward normalization 6 weeks after the suspension of fish oil supplementation. These effects of fish oil extracts and EPA on phagocytosis and humoral responses may be advantageously used in the therapy of chronic inflammatory diseases and autoimmune diseases but could be a cause for concern when these compounds are used for longer periods of time and with minimal medical supervision for the prophylaxis of atherosclerosis. PMID- 2544326 TI - Enalapril treatment of a nursing with slightly impaired renal function. PMID- 2544328 TI - Genital human papillomavirus infection. PMID- 2544327 TI - Neuropsychiatric abnormalities in primary Sjogren's syndrome. AB - Neuropsychiatric abnormalities were searched for in 52 patients with primary Sjogren's syndrome (SS). 40 patients were evaluated with a detailed neurologic history and physical examination and determination of terminal latencies and nerve conduction velocities of all limbs. Nineteen of them, plus another 12 primary SS patients were evaluated for hostility structure and psychiatric symptoms, using the Hostility and Direction of Hostility Questionnaire (HDHQ) and the Symptoms Check List-90R (SCL-90R) respectively and the results were compared with those of 33 healthy women and 41 cancer patients. Three patients had face numbness and/or hypesthesia indicating trigeminal involvement. Ten had mild sensory or mixed neuropathy of the glove-stocking type. None of our patients volunteered peripheral nervous system (PNS) symptoms nor did we detect central nervous system (CNS) involvement in any of them. One patient with purpura, glomerulonephritis and cryoglobulinemia presented with severe mononeuritis multiplex. High levels of introverted hostility were reported by SS patients in relation to the other two groups. In addition, higher scores on paranoid ideation, somatization and obsessive compulsiveness were found in SS patients compared to the rest but no correlation was found between psychiatric symptomatology and neurologic abnormalities. It is suggested that PNS disease is relatively common and benign in most primary SS patients, psychiatric disorders, sometimes serious, are also common, but CNS involvement must be rather rare. PMID- 2544329 TI - Milestones in HPV research. PMID- 2544330 TI - Epidemiologic aspects of genital HPV infection. PMID- 2544331 TI - Virology of human papillomavirus. PMID- 2544332 TI - Diagnosis of HPV infection by recombinant DNA technology. PMID- 2544333 TI - Linking HPV to cancer. PMID- 2544334 TI - Microscopic diagnosis of HPV infection. PMID- 2544335 TI - HPV-associated lesions of the cervix: biology and colposcopic features. PMID- 2544336 TI - Genital HPV infections in men. PMID- 2544337 TI - HPV-associated lesions in pregnancy and their clinical implications. PMID- 2544338 TI - Genital human papillomavirus infection. Management strategies. PMID- 2544339 TI - Extensive extraosseous localization of bone imaging agent in a patient with renal failure and rhabdomyolysis accompanied by combined hypercalcemia and hyperphosphatemia. AB - Four sequential Tc-99m pyrophosphate (PYP) imaging studies were performed in a 28 year-old man with high fever and exudate pharyngitis associated with renal failure. Radiotracer localization in the left ventricle (LV), lungs, kidneys, and skeletal muscles were seen in two, initial imaging studies. In the second and third imaging studies, area of increase in activity was seen in the left-sided bowel. In studies done two months later (in the third study), the radioactivity in the skeletal muscles was no longer seen. Studies obtained nine months (in the fourth study) after the first imaging showed less radiotracer localization in the LV, lungs, and kidneys as compared to that seen in the initial study. Myocardial necrosis and microcalcification were proved by LV biopsy. The exact mechanism of extraosseous bone-imaging agent localization is unknown. However, this phenomenon may be related to renal failure, rhabdomyolysis, hypercalcemia, hyperphosphatemia, or elevated parathyroid hormone. The Tc-99m PYP imaging study is useful and sensitive in the detection of extraosseous tissue calcification and monitoring of the disease process. PMID- 2544340 TI - Radionuclide detection of abnormal ventricular filling patterns in rejecting human allografts. AB - Parameters of systolic and diastolic function obtained from radionuclide ventriculography (RNV) were evaluated in nine cardiac allograft recipients. In 25 examinations, left end-diastolic volume (LEDV), cardiac output (CO), left ejection fraction (LEF), right ejection fraction (REF), heart rate (HR), peak filling rate (PFR), time to peak filling rate (TPFR), peak ejection rate (PER), and average filling rate for the first half of diastole (DFRH) were determined. Endomyocardial biopsy was obtained within 48 hours. Biopsies were divided into three treatment classes (0 = normal; 1 = rejection but not requiring supplemental therapy; and 2 = rejection requiring supplemental immunotherapy). Two independent variables of diastolic function proved to be significant (DFRH P less than 0.00001, and PFR P less than 0.002) predictors of the dependent variable class when regression analysis was applied to the data. Alterations in diastolic function associated with acute rejection are detectable on RNV and simulate changes anticipated in a primary restrictive cardiomyopathy. PMID- 2544341 TI - Technetium-99m pentavalent DMSA imaging detects metastases of poorly differentiated carcinoma of the thyroid. PMID- 2544342 TI - A direct method for the diagnosis of human hepatic type 1b and type 1c glycogen storage disease. AB - 1. Type 1b and type 1c glycogen-storage disease are caused respectively by deficiencies of the glucose-6-phosphate translocase and the phosphate/pyrophosphate translocase of the human hepatic microsomal glucose-6 phosphatase system. 2. Current methods of unequivocally diagnosing type 1b and type 1c glycogen storage disease are indirect and complex. 3. We have therefore developed a simple, rapid and direct microfiltration assay for the glucose-6 phosphate translocase and the phosphate/pyrophosphate translocase. 4. We have demonstrated that the microfiltration assay can be used to directly diagnose type 1b and 1c glycogen-storage disease in microsomes isolated from hepatic needle biopsy samples. PMID- 2544343 TI - Effect of nedocromil sodium on down-regulation of pulmonary beta-adrenoceptors. AB - 1. We have studied the effect of the anti-inflammatory anti-asthma drug, nedocromil sodium, on down-regulation of pulmonary beta-adrenoceptors in guinea pig lung. 2. Incubation of minced lung with isoprenaline (10 mumol/l) resulted in a reduction in maximum binding capacity of [125I]iodocyanopindolol to lung membranes from 246 +/- 4 to 169 +/- 6 fmol/mg of protein (mean +/- SEM, P less than 0.01, n = 18). 3. Nedocromil sodium, which had no direct effect on [125I]iodocyanopindolol binding, prevented isoprenaline-induced down-regulation, giving complete protection at a dose of 100 mumol/l. 4. The mechanism of this effect is not certain, but nedocromil sodium may interfere with the internalization of beta-adrenoceptors in pulmonary parenchymal cells. This may have some therapeutic relevance. PMID- 2544344 TI - Effects of pimobendan, a novel inotropic agent, on intracellular calcium and tension in isolated ferret ventricular muscle. AB - 1. In this study we have investigated the effects of a novel inotropic agent, pimobendan (UDCG 115-BS), on skinned and intact ventricular muscle from ferrets. 2. Pimobendan (20 or 100 mumol/l) increased tension at a given free [Ca2+] when applied to skinned ventricular muscle, i.e. it increased the Ca2+ sensitivity of the myofibrils. 3. Tension and intracellular free Ca2+ [( Ca2+]i) were measured simultaneously in intact papillary muscles using the aequorin technique. When 25 mumol/l pimobendan was added to the superfusing solution, a slowly developing positive ionotropic effect was produced, which was accompanied by an increase in the size of the systolic rise in [Ca2+]i (Ca2+ transients) with a similar time course. 4. In order to determine whether pimobendan increased the Ca2+ sensitivity of myofibrils in an intact papillary muscle, we compared the increase in Ca2+ transients and tension observed in response to changes in extracellular [Ca2+] with those observed in response to pimobendan. The result of this comparison was that in intact muscle pimobendan caused no apparent increase in myofibrillar Ca2+ sensitivity. 5. Pimobendan caused an abbreviation of the time course of the Ca2+ transients, but the twitch was slightly prolonged. 6. When isoprenaline was added to the superfusing solution, a positive inotropic effect was produced, which was accompanied by a marked increase in the size of the Ca2+ transients. Isoprenaline caused an abbreviation of the time course of both the Ca2+ transients and the twitch. When the Ca2+ sensitivity of the intact myofibrils was determined as described above, isoprenaline caused a desensitization. Pimobendan produced a sensitization when compared with isoprenaline. 7. These results are consistent with the hypothesis that pimobendan produces an inotropic effect in isolated cardiac muscle which is mediated both by an increase in Ca2+ sensitivity and by an increase in adenosine 3':5'-cyclic monophosphate due to its phosphodiesterase-inhibiting activity. Such a combination of activities may be particularly advantageous for an inotropic agent. PMID- 2544345 TI - Differential effect of high-dose naloxone on the plasma adrenaline response to the cold-pressor test. AB - 1. Although the opiate antagonist naloxone has been shown to affect sympathoadrenomedullary function in some studies, this has not been a uniform finding in all investigations, using different doses of naloxone. We have therefore investigated the actions of saline placebo and increasing bolus doses of intravenous naloxone (25, 100 and 250 micrograms/kg) on the plasma noradrenaline, adrenaline, adrenocorticotrophin (ACTH) and cortisol responses to a cold-pressor test in six males and two females in a double-blind randomized study. 2. Basal levels of adrenaline did not differ on any of the study occasions: the cold-pressor stimulus produced a significant rise in mean plasma adrenaline to a peak of 0.16 nmol/l, with a peak incremental rise of 0.08 nmol/l. In the presence of the two higher doses of naloxone, the incremental rise in the mean adrenaline level was significantly enhanced, reaching 0.30 nmol/l at 100 micrograms of naloxone/kg and 0.29 nmol/l at 250 micrograms of naloxone/kg, with no significant enhancement observed at the lowest dose of naloxone. The rise in plasma noradrenaline, systolic and diastolic blood pressure and pulse rate during the cold-pressor test was not consistently altered by any dose of naloxone, but there was a significant trend for the degree of discomfort to increase with the dose of naloxone. 3. Neither plasma ACTH nor serum cortisol rose in response to the cold-pressor stimulus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544346 TI - In vitro and in vivo phosphorylation of chicken beta B3-crystallin. AB - Incubations of chicken lens homogenates with [32P]-ATP revealed the phosphorylation of a 28 kDa protein, and phosphoamino acid analysis of the phosphorylated protein showed the presence of phosphoserine. The protein is present in the beta-crystallin fraction and after purification and partial sequence determination, by way of peptide mapping and subsequent amino acid analyses and Edman degradation, this 28 kDa protein was identified as the beta B3 crystallin subunit, based on its homology with the bovine and rat orthologue. From phosphate content determination it could be concluded that this chicken beta B3 subunit contains in vivo 2 mol phosphate/mol polypeptide. PMID- 2544347 TI - Benign liver tumors. AB - This article discusses the most important benign liver tumors, both in adult and pediatric patients. A pathologic discussion of each neoplasm is included to provide a basis for understanding the radiologic-pathologic correlation that is used throughout the monograph. The benign liver tumors are presented according to their frequency. Therefore, hemangioma, the most common primary benign liver neoplasm, is discussed first, followed by focal nodular hyperplasia, hepatocellular adenoma, and the benign primary pediatric tumors--infantile hemangioendothelioma and mesenchymal hamartoma. Finally, a brief discussion of nodular regenerative hyperplasia and other rare hepatic masses is included. Bile duct cyst (simple, non-parasitic cyst of the liver) is not included since it is not a neoplasm. Likewise, cystadenoma is not discussed since it originates from the biliary duct cell and is appropriately included in the biliary neoplasms category. PMID- 2544348 TI - Malignant liver tumors. AB - In this monograph we have presented the most important liver tumors in children and adults with pathological correlation. We believe that this information may be useful for understanding the varying radiological findings in malignant liver tumors since it includes pathological findings responsible for the radiographic images. It is clear that there are no characteristic signs that could establish a specific radiological diagnosis in malignant liver tumors. However, careful analysis of the radiographic images provided by different modalities and correlations with the clinical data may, in many cases, suggest a correct preoperative diagnosis. PMID- 2544349 TI - Neoadjuvant vindesine, etoposide, and cisplatin for locally advanced non-small cell lung cancer. Final report of a phase 2 study. AB - We treated 27 patients with regionally advanced non-small-cell lung cancer (NSCLC) with two cycles of neoadjuvant chemotherapy with etoposide, vindesine, and cisplatin. Twenty-three patients were evaluable for response; 13 had a partial response while ten patients had stable disease or disease progression. Subsequent local therapy consisted of surgery followed by radiotherapy in four patients and of radiotherapy alone in 14 patients. Five patients did not receive local therapy. At completion of local therapy, seven patients were considered free of disease including all four who had undergone surgery. Median time to disease progression for the 13 patients who had a partial response to neoadjuvant chemotherapy was eight months (three to 51+ months). The median survival for all patients registered on study was eight months (three days to 53+ months). Chemotherapy induced toxicities included moderate myelosuppression, nausea and vomiting in all patients, and occasional ototoxicity, neurotoxicity, and wasting syndrome. One patient died of intracerebral hemorrhage due to thrombocytopenia. This trial shows that administration of neoadjuvant chemotherapy to patients with locoregionally advanced NSCLC is feasible and may yield an increased response rate compared to patients with stage IV disease. While no clearly beneficial effect of the use of chemotherapy on patient survival is apparent in this study, further studies utilizing neoadjuvant chemotherapy in patients with NSCLC are warranted and should attempt to identify more active combinations of drugs. PMID- 2544350 TI - Pulmonary complications in lead miners. AB - We carried out a study to assess the prevalence of respiratory disease in lead miners and to investigate the roles of silica and lead. We used a questionnaire for symptoms and examinations for signs of respiratory disease, chest roentgenograms, and spirometric study in 45 lead miners. Six underwent bronchoscopy and transbronchial lung biopsy (TBB) and five lung lead analysis. Lung lead levels from five patients with no occupational lead exposure were obtained for comparison. Results showed restriction in five of 45 and reticulonodular opacities in 16 of 45 workers. Squamous metaplasia and other histopathologic changes were observed, although silicotic nodules were absent by TBB. Lung lead levels above those of control subjects were observed in four of five lead miners. These findings show that lead miners are at risk for lung disease. Although silica is a likely cause, elevated lung lead content found in these miners merits further investigation. PMID- 2544351 TI - Gallium lung scanning and bronchoalveolar lavage in crocidolite-exposed workers. AB - Gallium lung scanning is widely used to evaluate pulmonary inflammation in patients with interstitial lung disease but has not previously been reported in crocidolite-exposed workers. In order to characterize the pulmonary inflammation caused by crocidolite inhalation, GLS and BAL findings were related to chest x ray film changes graded according to the ILO classification of roentgenograms of pneumoconioses. In individuals with roentgenographic evidence of asbestosis (CXR greater than or equal to 1/0, n = 15), 13 had a positive GLS and 13 had an abnormal BAL. In asbestos-exposed individuals with equivocal chest x-ray film changes (CXR 0/1, n = 12), six had a positive GLS and six had BAL changes (both GLS and BAL abnormal in three). In individuals with a normal chest x-ray film (CXR 0/0 n = 8), two had a positive GLS and two BAL changes (both abnormal in 1). These data demonstrate that most subjects with crocidolite-induced asbestosis have an abnormal GLS and BAL. In addition, many individuals with asbestos exposure and equivocal or no chest x-ray film changes have an abnormal GLS and/or BAL, suggesting the presence of active subclinical pulmonary inflammation in these individuals. PMID- 2544352 TI - Comparison of antihypertensive therapies by noninvasive techniques. AB - We compared the antihypertensive effects of the beta-blocker atenolol and the converting enzyme inhibitor lisinopril during 12 weeks of treatment in patients with mild to moderate essential hypertension. Atenolol (n = 10) significantly decreased conventionally measured blood pressure from 144/103 to 135/93 mm Hg and lisinopril (n = 9) from 150/104 to 130/92 mm Hg. Based on data derived from automated 24-h ambulatory blood pressure monitoring, atenolol decreased the average whole-day systolic pressure by 18 +/- 6 mm Hg (p less than 0.02) and the diastolic pressure by 11 +/- 2 mm Hg (p less than 0.01). Lisinopril produced decreases of 27 +/- 5 mm Hg (p less than 0.01) and 13 +/- 2 mm Hg (p less than 0.001). Examination of the 24-h blood pressure patterns showed that the efficacies of the two drugs were similar. Each appeared to be effective throughout the whole-day monitoring period, although only lisinopril significantly decreased blood pressure during the final four-h period (4 AM to 8 AM) preceding the next day's dose. Neither drug produced significant echocardiographic changes in left ventricular wall thickness or muscle mass during the short-term treatment. Lisinopril and atenolol effectively decrease blood pressure during a 24-h period. Moreover, we found that automated whole-day blood pressure monitoring is a useful tool for comparing the efficacy and duration of action of differing antihypertensive agents. PMID- 2544353 TI - Quality of life assessment in patients with carcinoma of the lung. PMID- 2544354 TI - Oncogenes and genetic abnormalities in lung cancer. PMID- 2544357 TI - The clinical relevance of recent developments in pathology and biology of small cell lung cancer. PMID- 2544356 TI - Lung cancer and autocrine growth factors. PMID- 2544355 TI - Monoclonal antibodies in lung cancer. PMID- 2544358 TI - Advances in the biology of lung cancer. Clinical significance of neuroendocrine differentiation. PMID- 2544359 TI - Radiologic staging of lung cancer using CT and MRI. PMID- 2544360 TI - Value of the new TNM staging system for lung cancer. PMID- 2544361 TI - Limited resection in the treatment of stage I non-small cell lung cancer; an overview. PMID- 2544362 TI - Innovative chemotherapy. Xenografts and in vitro drug sensitivity testing. PMID- 2544363 TI - New treatments for small cell lung cancer. When to test. PMID- 2544364 TI - Chemotherapy of small cell lung cancer. AB - Selection of appropriate treatment is now possible on the basis of prognostic indices. For patients with a "poor" prognosis, therapy should be minimally toxic, for palliative purposes only. For patients with a "good" prognosis, intensive treatment is recommended with combinations comprised from C, A, V, VP-16, and CP. For patients obtaining a remission, consolidation is recommended with radiation treatment if this has not been part of the initial induction program with or without chemotherapy. Consolidation may be intensified by using high-dose chemotherapy in association with autologous bone marrow transplantation or possibly the use of hematopoietic growth factors. The major problem limiting further improvements in survival in this disease remains the emergence of drug resistance, which is now the subject of intensive investigations both in the laboratory and in the clinic. PMID- 2544365 TI - Optimum duration of treatment with combination chemotherapy for small cell lung cancer. PMID- 2544366 TI - Should we intensify the treatment of small cell lung cancer? PMID- 2544367 TI - High-dose chemotherapy in lung cancer. PMID- 2544369 TI - Hyperthermia in the management of lung cancer. The current situation. PMID- 2544368 TI - Altered fractionation for non-small cell carcinoma of the lung. Rationale for the prospective trials of the Radiation Therapy Oncology Group. PMID- 2544370 TI - High-LET radiation therapy of non-small cell lung cancer. PMID- 2544371 TI - Report on the IASLC Le Havre Workshop on combined modality treatment in small cell lung cancer. PMID- 2544372 TI - Combined modality treatment of small cell lung cancer. PMID- 2544373 TI - How should thoracic radiotherapy be given in limited small cell lung cancer? PMID- 2544374 TI - Summary of pathobiologic presentations. PMID- 2544375 TI - Report of the IASLC Le Havre Workshop. PMID- 2544376 TI - Treatment of N2 non-small cell lung cancer (NSCLC). PMID- 2544377 TI - Radiotherapy in non-small cell lung cancer. An overview. PMID- 2544378 TI - Adjuvant therapy for non-small cell lung cancer. PMID- 2544379 TI - The epidemiology and pathogenesis of malignant mesothelioma. PMID- 2544380 TI - [Urachus cancer--a rare malignant form of urachus abnormality]. PMID- 2544381 TI - [Effect of pyrethroids on rat brain synaptosomal ATPase activities]. AB - In vitro Effect of several pyrethroids on rat brain synaptosomal ATPase activities was investigated. No significant changes in Na+, K+-ATPase and oligomycin-insensitive Mg2+-ATPase activities were observed under present experimental conditions, but all pyrethroids tested caused significant inhibition of oligomycin-sensitive Mg2+-ATPase activity with certain concentration dependence. The results suggest a possibility that pyrethroids may alter the cellular energy metabolism of the nervous system. PMID- 2544382 TI - New approach to surgical treatment of ulcerative colitis and polyposis coli without pelvic pouch. Experimental study in dogs. AB - A new method for the surgical treatment of ulcerative colitis and polyposis coli is described. Instead of preparing a pelvic pouch, the natural rectal pouch stripped of the diseased mucosa was used experimentally in dogs. The undisturbed muscular cuff of the rectum (12 cm from the anal verge) was covered by healthy vascularized mucosa of small bowel in such a manner that the dog could use its rectum as usual before surgery. The results are encouraging. The rectal reservoir is spared, with its sensitivity, continence and motor activity covered by healthy mucosa. The dogs thrived. PMID- 2544383 TI - Multiple adenomas in terminal ileum 25 years after restorative proctocolectomy for familial adenomatous polyposis. Report of a case. AB - A patient with familial adenomatous polyposis was treated with colectomy, mucosal proctectomy, and a straight ileoanal anastomosis in 1962. Thirteen to 21 years later recurrent adenomas developed at the ileoanal anastomosis, and 25 years after the operation multiple adenomas were found in the terminal ileum up to 12 cm from the ileoanal anastomosis. It is concluded that colectomy and mucosal proctectomy, with or without an ileoanal reservoir, does not eliminate the future risk of adenoma formation. Thus, this method should be used only in selected polyposis patients. PMID- 2544384 TI - Familial adenomatous polyposis. PMID- 2544385 TI - [Transposon-like DNA sequence encoding neomycin resistance in Streptomyces lividans 66]. PMID- 2544386 TI - [Structure of a new transformation-defective mutant of Rous sarcoma virus]. PMID- 2544388 TI - [Highly repetitive sequences in the genome of the rat-like hamster Tscherskia triton are represented by two families of dispersed ubiquitous repeats]. PMID- 2544387 TI - [Stimulation of phosphoinositide turnover is an important but not a sole condition for activation of human T-lymphocyte proliferation by phytohemagglutinin and interleukin 2]. PMID- 2544389 TI - [Strains of Bacillus thuringiensis subspecies produce two delta- endotoxins in various ratios]. PMID- 2544390 TI - Some silica-based double salts as ion exchangers useful in the treatment of industrial effluents and control of pollutants. AB - A systematic comparative study has been performed on the various double salts based on silica. They are found to be more stable chemically and thermally than single salts of this type. Furthermore, these materials have a better selectivity for some pollutants chiefly found in the effluents of dyestuff and plating industries. The purpose of preparing a series of such materials in this study has been to evaluate their comparative usefulness under the varying conditions of pollution. The materials have been characterized on the basis of analytical studies such as chemical composition, pH titration, IR, X-ray, and TGA. PMID- 2544391 TI - [Carcinogenic properties of silicon carbide whiskers]. AB - In experiments with intrapleural injections of silicon carbide whiskers (20 mg X 3, with one month interval) to random-inbred rats their carcinogenic activity close to that of chrysotile B UICC has been established. The pleural mesotheliomas were induced in 47.7 and 34.1% of rats, respectively. PMID- 2544392 TI - [The activity of nuclear endonucleases and topoisomerases in the liver of rats and in diethylnitrosamine-induced tumors]. AB - The DNA endonuclease (Aendo) and DNA topoisomerase (Atopo) activities in liver nucleus extracts of normal rats, in DENA-induced hepatomas and in liver tissues around tumours were investigated. The profile of nuclear endonucleases measured in the presence of 2 mM CaCl2 + 5 mM MgCl2, or 5 mM MnCl2, or 5 mM MgCl2, or 2 mM CaCl2 (pH 7.4), or I mM EDTA (pH 5.0) was different in normal and tumour tissues. Mn2+-dependent endonuclease was the main endonuclease in the tumour tissue, whereas Ca2+, Mg2+-dependent endonuclease was the main one in the normal liver and in the tissue around the tumour. An increase in the Mn2+-dependent endonuclease activity correlated with a decrease in the hepatoma differentiation level. Atopo of types I and II increased in the tissue around the tumour. Aendo and Atopo of cellular nuclei decreased in animals given DENA without the liver tumour. PMID- 2544393 TI - [Interaction of the ligand molecules with the membrane receptors of tumor cells]. AB - The mechanism of the ligand molecule interaction with cell membrane receptors is considered. The equation is obtained which describes dynamics of change in the number of ligand-receptor complexes on membranes with their constant internalization. It is shown that the in vivo internalization leads to saturation of the cell surface with the ligand-receptor complexes, the ligand concentrations being less than Kd. Theoretical conclusions are compared with the experimental data. PMID- 2544394 TI - [Quantitative evaluation of the surface membrane receptors of leukemic cells to blood serum transcobalamin-II]. AB - Serum protein transcobalamin II (TC-II) is responsible for transport of cobalamins into mammalian cells. A method of quantitative estimation of plasma membrane receptors of hemopoietic cells to TC-II cobalamin complex is suggested. Analysis of mouse leukemia L1210 cells includes the saturation of radiolabelled ligand-receptor complex with papain. The number of receptors and 57CoCNCbl content in one cell is determined by differentiated radioactivity count of solubilized protein complexes and of cytoplasm. PMID- 2544395 TI - [Human papillomatosis viruses and cervical cancer]. AB - Recent data on the role of human papillomaviruses in the arising and development of cervical neoplasia are reviewed. Cervical dysplasia is characterized, its natural history and the role of papillomaviruses in arising of dysplasia and cervical cancer are described. Modern results on identification of the capsid antigen and DNA of human papillomaviruses in different cervical lesions are presented. Several types of papillomaviruses with different oncogenic potential, as well as molecular aspects of the virus gene expression and cellular differentiation are discussed. PMID- 2544396 TI - Extracellular adenosine triphosphate completely reverses the thyrotropin-induced morphological change in FRTL-5 cells. AB - We quantified the TSH-induced morphological change in FRTL-5 thyroid cells according to a morphological index corresponding to the mean cell area measured from microscopic photographs. Within 15 min, TSH induced, at 10 pM and higher concentrations, a decrease in morphological index together with a rise in cAMP levels in a TSH dose-dependent manner. Forskolin, 3-isobutyl-1-methylxanthine, and RO 20-1724, the latter two being phosphodiesterase inhibitors, mimicked these TSH effects, indicating that the rise in cAMP levels is responsible for the TSH effect. Extracellular ATP and its derivatives, known as purinergic receptor agonists, decreased cAMP levels and caused a complete reversal of the TSH morphological effect. Prior exposure of the cells to islet-activating protein (pertussis toxin), the depletion of extracellular Ca2+, or the addition of low doses of protein kinase-C inhibitors completely abolished the inhibitory action of ATP on the TSH effect, whereas phorbol 12-myristate 13-acetate, which activates protein kinase-C, mimicked the ATP action to some extent. Thus, although the TSH-induced change in cell morphology seems to be dependent on cAMP levels, the inhibition of TSH action by ATP seems to be mediated by at least two signal transduction pathways involving islet-activating protein substrate G proteins: one inhibiting adenylate cyclase and the other involving Ca2+ and protein kinase-C. PMID- 2544397 TI - Epidermal growth factor stimulates tissue plasminogen activator activity and messenger ribonucleic acid levels in cultured rat granulosa cells: mediation by pathways independent of protein kinases-A and -C. AB - Recent reports suggest that epidermal growth factor (EGF) or related peptides may act as local hormones to regulate granulosa cell differentiation. While FSH and GnRH are known to stimulate accumulation of tissue-type plasminogen activator (tPA) mRNA in granulosa cells, studies using nonovarian cells have shown stimulation of tPA by EGF. In this study, the effect of EGF and its structural analog transforming growth factor-alpha (TGF alpha) on ovarian tPA mRNA and activity was investigated. Granulosa cells obtained from immature estrogen treated rats were cultured with FSH or increasing doses of EGF or TGF alpha before analysis of tPA activity using sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by a fibrin overlay technique. Like FSH and GnRH, EGF and TGF alpha stimulated the secretion of tPA activity in a dose- and time dependent manner (onset, 12 h; maximum, 48 h). Northern blot hybridization of total RNA using a rat cRNA probe for tPA showed the accumulation of a 22S species mRNA in cells treated with EGF or TGF alpha, but not with nerve growth factor, suggesting increased expression of the tPA gene. Furthermore, slot blot hybridization of RNA from these cells confirmed a time-dependent increase in tPA mRNA preceding that in enzyme activity. Cotreatment of a saturating dose of EGF with phorbol myristate acetate (PMA) or GnRH resulted in additive increases in both tPA enzyme activity and mRNA levels. In addition, pretreatment with PMA desensitized the cells to subsequent treatment with PMA or GnRH, but did not diminish EGF-induced tPA mRNA, suggesting that EGF acts through a pathway independent of protein kinase-C. Also, extracellular cAMP levels did not increase with EGF treatment in the presence or absence of a phosphodiesterase inhibitor, suggesting the lack of involvement of the protein kinase-A pathway. Suppression of protein synthesis by cycloheximide inhibited the induction of tPA mRNA by EGF, whereas similar treatment resulted in the superinduction of tPA mRNA in FSH treated cells, suggesting that EGF and FSH do not share the same pathway. These results suggest that EGF and TGF alpha induce tPA mRNA and activity in granulosa cells through a pathway independent of protein kinases-A (FSH) and -C (GnRH and phorbol ester), providing an interesting model for future elucidation of the molecular mechanism involved in tPA gene expression. PMID- 2544398 TI - Inhibitory effects of interleukin-1 on luteinizing hormone-stimulated adenosine 3',5'-monophosphate accumulation by cultured porcine granulosa cells. AB - To elucidate the mechanisms of the inhibitory effect of interleukin-1 (IL-1) on LH-stimulated progesterone secretion by cultured porcine granulosa cells, we examined which steps of the LH-stimulated, cAMP-mediated, progesterone biosynthetic pathway were affected by IL-1. Pretreatment of the cells for 48 h with IL-1 reduced intra- and extracellular cAMP accumulation in response to LH by 73% and 83%, respectively. The inhibitory effects of IL-1 were time and concentration dependent. Significant inhibition was observed at as low as 50-250 pg/ml, and the effect was maximal at 100 ng/ml (ID50; 2 ng/ml). The lowest concentration of IL-1 used (0.5 pg/ml), in contrast, showed a tendency to stimulate LH-induced cAMP accumulation. Effect of IL-1 on the specific binding of [125I]LH to granulosa cells was then examined, which showed that IL-1 (100 ng/ml) significantly reduced the specific binding by 36%. IL-1 also significantly reduced intra- and extracellular cAMP accumulation by the cells in response to forskolin (50, 150 microM) by 28-46%, indicating that IL-1 can directly inhibit adenylate cyclase activity. Contrary to these inhibitory actions of IL-1 on LH stimulated cAMP generation, IL-1 did not significantly reduce progesterone secretion induced by (Bu)2cAMP. These results indicate that the inhibitory effect of IL-1 on LH-stimulated progesterone secretion is due to its actions at at least two different sites along the LH-stimulated, cAMP-mediated, progesterone biosynthetic pathway, the LH receptor level and adenylate cyclase systems. The post-cAMP steps of progesterone production, in contrast, did not seem to be affected by IL-1. PMID- 2544399 TI - The antigonadotropic action of prostaglandin F2 alpha is not mediated by elevated cytosolic calcium levels in rat luteal cells. AB - We have investigated the role of intracellular calcium in the mechanism of action of prostaglandin F2 alpha (PGF2 alpha) in cultured rat luteal cells. PGF2 alpha (1 microM) maximally inhibited LH-stimulated cAMP accumulation and also initiated a transient release of intracellular calcium. Low doses of the calcium ionophore ionomycin also increased intracellular calcium to a similar extent as PGF2 alpha (1 microM), but did not inhibit LH-stimulated cAMP accumulation. Chelation of intracellular calcium with dimethyl bis-(o-aminophenoxy)ethane-N,N,N',N' tetraacetic acid (BAPTA) (10 microM) attenuated the transient calcium rise stimulated by PGF2 alpha, but did not affect the inhibitory characteristics of PGF2 alpha on LH-stimulated cAMP accumulation. Treatment of luteal cells with EGTA (1 mM) and ionomycin (500 nM) resulted in depletion of intracellular calcium to such an extent that a subsequent exposure of the luteal cells to PGF2 alpha (1 microM) did not elicit any change in intracellular calcium. Depletion of intracellular calcium and ablation of the calcium response to PGF2 alpha, however, did not affect either the dose response or the time course of inhibition of LH-stimulated cAMP accumulation. We conclude that although intracellular calcium is mobilized by PGF2 alpha in cultured rat luteal cells, the antigonadotropic action of PGF2 alpha on LH-stimulated cAMP accumulation is not mediated by this mechanism. PMID- 2544400 TI - The enhancing effect of adrenocorticotropin on adrenal 3-hydroxy-3-methylglutaryl coenzyme A reductase messenger ribonucleic acid level is inhibited by aminoglutethimide but not by cycloheximide. AB - 3-Hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) reductase activity and reductase mRNA level were determined in adrenals from hamsters treated with ACTH, with or without cycloheximide or aminoglutethimide. Both reductase activity and reductase mRNA level were similarly enhanced by ACTH administration compared to levels in NaCl-treated animals. The administration of cycloheximide with ACTH resulted in a 73% decrease in reductase activity compared to control values, but did not prevent the enhancing effect of ACTH on the reductase mRNA level. Furthermore, the administration of cycloheximide alone diminished HMG-CoA reductase activity, but enhanced by 1.1- to 1.6-fold the reductase mRNA level. Coadministration of aminoglutethimide with ACTH also resulted in a decrease (65%) in reductase activity compared to that in NaCl-treated animals. However, coadministration of aminoglutethimide, in contrast to cycloheximide, with ACTH not only prevented the reductase mRNA level increase produced by ACTH, but also resulted in a 30% decrease in the reductase mRNA level compared to that in controls injected with 0.15 M NaCl. In addition, aminoglutethimide alone resulted in 50% and 54% decreases in reductase mRNA level and reductase activity, respectively. Thus, we have shown that both cycloheximide and aminoglutethimide can prevent the enhancing effect of ACTH on HMG-CoA reductase activity, but their modes of action differ. It is likely that the aminoglutethimide inhibition could be the result of a diminution of specific reductase gene transcription, whereas cycloheximide would result in inhibition of the synthesis of specific proteins, including HMG CoA reductase. In this respect, since the adrenal free cholesterol content was increased in groups treated with ACTH-aminoglutethimide, we postulate that free cholesterol could be one of the important components involved in the regulation of HMG-CoA reductase gene transcription. As for the ACTH-cycloheximide-treated groups, the adrenal free cholesterol content was also increased, but the effect of ACTH on the reductase mRNA level was not prevented, presumably because this drug blocked the synthesis of a putative sterol regulatory protein that is required to repress HMG-CoA reductase gene transcription. PMID- 2544401 TI - Parathyroid hormone (PTH)-related protein is a potent stimulator of osteoclast like multinucleated cell formation to the same extent as PTH in mouse marrow cultures. AB - Induction of osteoclast-like multinucleated cells (MNCs) by various fragments of PTH-related protein (PTHrP) was examined in mouse marrow cultures. Osteoclast like MNCs were defined as tartrate-resistant acid phosphatase (TRACP)-positive MNCs with calcitonin receptors. In all experimental protocols examined, PTHrP-(1 34) induced TRACP-positive MNCs at almost the same rate as PTH-(1-34). PTHrP-(1 29) was less potent than PTHrP-(1-34). PTHrP-(1-25) and PTHrP-(1-14) had no effect. PTHrP-(1-34) was more potent than PTH-(1-34) in increasing the accumulation of cAMP, but the former appeared to lose its activity more rapidly than the latter. Isobutylmethylxanthine increased the effect of PTHrP-(1-34) and PTH-(1-34) in inducing TRACP-positive MNCs. Furthermore, the calcium ionophore A23187 significantly increased the formation of TRACP-positive MNCs. The effect of PTH-(1-34) and PTHrP-(1-34) in inducing TRACP-positive MNCs was potentiated by adding A23187 but suppressed by adding verapamil simultaneously. The inhibition by verapamil was overcome by adding A23187. [Nle8,18,Tyr34]PTH-(3-34)amide inhibited the effect of not only PTH-(1-34) but also PTHrP-(1-34) in inducing both the accumulation of cAMP and the TRACP-positive MNC formation. These results show that PTHrP is a potent stimulator of osteoclast-like MNC formation to almost the same extent as PTH. It increases the number of osteoclast-like MNCs by a mechanism involving cAMP and calcium ions, and is most likely mediated through the same receptor. The controversial results of the bone-resorbing activity of PTH and PTHrP reported so far may be explained by the differences in the relative potencies of the respective hormones in increasing the intracellular cAMP and calcium ions and by the shorter half-life of PTHrP in culture medium. PMID- 2544402 TI - A structurally unique, potent, and selective oxytocin antagonist derived from Streptomyces silvensis. AB - The in vitro and in vivo oxytocin/arginine vasopressin (OT/AVP) antagonist properties of two cyclic hexapeptides derived from a newly discovered natural product (L-156,373) of Streptomyces silvensis are described. In radioligand binding assays, L-156,373 [cyclo(L-Pro-D-Phe-N-OH-L-Ile-D-piperazyl-L-piperazyl-N Me-D -Phe)] exhibited moderate affinity for rat uterine OT receptors (Ki, 150 nM), with some selectivity (approximately 20-fold) vs. liver AVP-V1 and kidney AVP-V2 receptors. Dehydroxylation of N-hydroxyisoleucine and oxidation of the piperazic acid residues of L-156-373 produced an interesting derivative, L 365,209. These structural modifications increased OT receptor affinity and selectivity by 20- and 2.5-5-fold, respectively. In the isolated rat uterus, L 365,209 was a potent (apparent dissociation constant, 1.7 nM) and competitive OT antagonist. L-365,209 also blocked the effects of AVP at both AVP-V1 (phosphatidylinositol turnover in rat hepatocytes) and AVP-V2 (adenylate cyclase in rat kidney medulla) receptors, but only at low micromolar concentrations. L 365,209, given iv to anesthetized rats, antagonized the action of exogenous OT on the uterus (ID50, 460 micrograms/kg) with a relatively long duration of action. L 365,209 represents a unique class of compounds that provides an entirely new approach for the design of antagonists for these neurohypophyseal hormones. PMID- 2544403 TI - Hypoglycemia enhances turnover of corticotropin-releasing factor and of vasopressin in the zona externa of the rat median eminence. AB - Insulin administration to overnight fasted rats causes a dose-dependent decline in plasma glucose concentrations and a dose-dependent increase in plasma ACTH concentrations. The ACTH response, but not the glucose response, was blocked by treatment with chlorpromazine-morphine-pentobarbital, indicating that the main factors triggering the ACTH response are of central, rather than peripheral, origin. To study whether insulin affected the turnover of CRF and vasopressin (AVP) in the zona externa of the median eminence (ZEME), we determined the rate of decline of both hypophysiotropic factors in rats with or without blockade of axonal transport by colchicine. In the ZEME, the concentrations of CRF and AVP were assessed by quantitative immunocytochemistry (QICC) in tissue sections or by RIA in median eminence extracts. QICC allows selective quantification of AVP and other peptides within the ZEME. The changes in the CRF content, as measured by QICC and RIA, were linearly correlated (r = 0.99), demonstrating that changes in peptide-staining intensity reflect changes in peptide content. Colchicine, when given intracisternally in a nontoxic dose of 5 micrograms, had no marked effect on resting plasma levels of ACTH and only slightly reduced the ACTH response to insulin-induced hypoglycemia. In the ZEME, CRF and AVP concentrations at rest were not affected by colchicine. In colchicine-treated rats insulin-induced hypoglycemia resulted in a prominent decline in CRF and AVP concentrations in the ZEME. The CRF concentration declined at a rate of 23%/h over a period of 3 h. The AVP concentration declined to a similar extent as CRF over the first hour, but tended to fall at the later time points. We conclude that hypoglycemia increases turnover of both CRF and AVP in the ZEME. However, the turnover rates of both hypophysiotropic peptides do not appear to be quantitatively coupled. PMID- 2544404 TI - Phosphoinositide hydrolysis and insulin secretion in response to glucose stimulation are impaired in isolated rat islets by prolonged exposure to the sulfonylurea tolbutamide. AB - Isolated rat islets of Langerhans were incubated for 2 h in a [3H]inositol containing medium supplemented with 7 mM glucose and the sulfonylurea tolbutamide (50-200 microM). After labeling, the ability of these islets to respond during a subsequent perifusion to 20 mM glucose or 15 mM alpha-ketoisocaproate (KIC) was assessed. The following major observations were made. Prior exposure to tolbutamide inhibited [3H]inositol efflux, inositol phosphate accumulation, and the insulin secretory responses of subsequently perifused islets to 20 mM glucose stimulation. When present during the 2-h labeling period, the calcium channel blocker nitrendipine (500 nM), a compound that abolishes tolbutamide-induced increases in PI hydrolysis, blocked these inhibitory effects of tolbutamide. In addition, the diacylglycerol kinase inhibitor monooleoylglycerol (50 microM) restored the impaired second phase insulin secretory response noted after a 2-h tolbutamide exposure. Prior exposure to tolbutamide (200 microM) also desensitized the islet, in terms of [3H] inositol phosphate accumulation, [3H]inositol efflux, and insulin secretory responses, to 15 mM KIC. The inclusion of monooleoylglycerol during the stimulatory period with KIC restored second phase insulin secretion. The results support the conclusion that chronic tolbutamide-induced increases in PI hydrolysis render the beta-cell insensitive to a subsequent 20-mM glucose or 15-mM KIC stimulus. Blocking tolbutamide-induced increases in PI hydrolysis during the labeling period eliminates the adverse effects of the sulfonylurea. The ineffectiveness of glucose and KIC to maintain insulin secretory responses from prior tolbutamide-exposed islets appears to be the result of the inability of these agonists to appropriately activate PI hydrolysis. PMID- 2544405 TI - Protein kinase-A and the effects of parathyroid hormone on phosphate uptake in opossum kidney cells. AB - Current evidence indicates that signal transduction after receptor binding of PTH involves the stimulation of adenylate cyclase as well as stimulation of phosphoinositide metabolism. Recent studies, showing that PTH alters phosphate transport in opossum kidney cells at concentrations which do not increase cAMP production and that activators of protein kinase-C also alter phosphate transport, have led to the suggestion that there is a dual mechanism for the regulation of phosphate transport by PTH, namely, protein kinase-C at physiological levels of PTH and cAMP at higher levels of PTH. The present studies were designed to evaluate the relationship between cAMP-dependent protein kinase (PK-A), a more sensitive indicator of alterations in cAMP metabolism than measurements of total cellular cAMP, and phosphate transport in opossum kidney cells, in response to bovine (b)PTH 1-34 and [Nle8,Nle18,Tyr34]bPTH 3-34 amide. While bPTH 1-34 markedly stimulated cAMP accumulation (half-maximal stimulation between 1 and 10 nM), PTH 3-34 analog did not. Phosphate transport was inhibited in a dose-dependent manner by bPTH 1-34, with half-maximal effect occurring between 0.1 and 1 nM. [Nle8,Nle18,Tyr34]bPTH 3-34 amide also altered phosphate transport, although this peptide was 3 orders of magnitude less potent than bPTH 1-34. PK-A activity increased in response to bPTH 1-34 and correlated closely with the effects of PTH on phosphate transport. [Nle8,Nle18,Tyr34]bPTH 3-34 amide, which did not appear to increase cAMP, also resulted in a significant increase in the activity of PK-A. Studies of inhibition of cAMP accumulation using 2',5'-dideoxyadenosine demonstrated that while this agent markedly inhibited the accumulation of cAMP in response to PTH, the effects of PTH on phosphate transport were not altered. However, in spite of the reduction in cAMP the activation of PK-A was similar to control. These data indicate that the effects of PTH peptides on phosphate transport are more closely related to changes in the activity of PK-A than to levels of total cAMP. Activation of PK-A in response to PTH is demonstrable at the lowest doses of PTH that alter phosphate transport. PMID- 2544406 TI - Rapid as well as delayed inhibitory effects of glucocorticoid hormones on pituitary adrenocorticotropic hormone release are mediated by type II glucocorticoid receptors and require newly synthesized messenger ribonucleic acid as well as protein. AB - Glucocorticoid hormones suppress the release of ACTH by the anterior pituitary gland: rapid feedback inhibits hormone secretion within 30 min of steroid application, delayed feedback is most effective at 1-2 h, and slow feedback becomes manifest in several hours. The aim of the present study was to determine the type of glucocorticoid receptor that mediates the rapid and delayed feedback actions of glucocorticoids and whether genomic activation occurs during the rapid and delayed time domains. Rat anterior pituitary cell columns were perfused with Dulbecco's minimum essential medium, 41-residue CRF (10(-9) M) was used as the secretagogue, which stimulated ACTH secretion to a peak of about 8- to 10-fold of basal release. The amount of ACTH released upon repeated 5 or 10 min stimulation with CRF was constant. Treatment with 10(-7) M corticosterone for 20 min immediately before and for 10 min during stimulation with CRF reduced ACTH release by about 50% (rapid feedback), while at 1 h and 2 h after the initial exposure to corticosterone the secretory response was 33% and 15% of control, respectively. The effect of corticosterone was prevented by the type II glucocorticoid/progesterone antagonist RU 38486 (10(-6) M). The selective type II receptor agonist RU 28362 (10(-7) M) was even more potent than corticosterone in inhibiting ACTH release; the time course of action was similar. When actinomycin D (10(-4) M) was applied in conjunction with RU28362 or corticosterone, no inhibitory effects appeared up to 2 h after the exposure to steroid. Puromycin (10(-4) M), given during and for 1 h after the administration of the steroid prevented the rapid as well as the delayed (1 h) inhibitory action of RU28362. When puromycin was removed from the system, a 75% inhibition of stimulated ACTH release developed at 2 h after the application of the steroid, indicating that translatable messenger RNA (mRNA) was still present in the cells. Cycloheximide (10(-4) M) was only partially effective at inhibiting rapid or delayed feedback, and increasing its concentration impaired the ACTH response to CRF-41. In summary, at the pituitary level the rapid as well as the delayed feedback inhibition of ACTH secretion by adrenal corticoids is exerted via type II glucocorticoid receptors. Furthermore, both rapid and delayed feedback require the synthesis of new mRNA and protein. PMID- 2544407 TI - Melatonin receptors in chick brain: characterization and localization. AB - Melatonin receptors in chick brain were characterized by RRA and localized by in vitro autoradiography, using [125I]melatonin [( 125I]MEL), a biologically active melatonin analog. In membranes from whole brain, radioreceptor studies revealed a high affinity [125I]MEL binding site with an equilibrium dissociation constant of 47.2 +/- 11.5 (mean +/- SEM) pM and a density of 37.8 +/- 8.5 fmol/mg protein. Binding was reversible and competitively inhibited by melatonin and closely related melatonin analogs, but not be norepinephrine or serotonin. In vitro autoradiographic studies of brain revealed a widespread distribution of melatonin receptors. Specific I-MEL binding was observed in retinorecipient and integrative nuclei of the visual system including the avian homolog of the mammalian suprachiasmatic nuclei. Auditory relay nuclei and limbic structures associated with arousal and vocalization were also specifically labeled. This widespread distribution of putative melatonin receptors in chick brain is in marked contrast to the very restricted distribution of melatonin receptors in mammalian brain and suggests that avian sensory systems are affected by melatonin. PMID- 2544408 TI - Pituitary immediate release pools of growth hormone and prolactin are preferentially refilled by new rather than stored hormone. AB - Pituitary stores of rat GH (rGH) and PRL (rPRL) are divisible into immediately releasable and more stable compartments representing either compartmentalized hormone within individual cells of a homogeneous population or responses of specialized cell subsets in a functionally heterogeneous population. In addition, newly synthesized rGH and rPRL can be processed either into intracellular storage or toward direct release. Fractional assignment of new hormone to these two paths can be influenced in the somatotroph by GHRH and may also represent either intracellular processes or functional heterogeneity of cells. We investigated the source, newly synthesized or stored, of hormone refilling the somatotroph and lactotroph immediately releasable pools (IRP) after their discharge by 21 mM potassium ion, 1 mM (Bu)2cAMP, 3 nM human GHRH-44, or 3 microM prostaglandin E1. Experiments were performed using perifused pituitary fragments exposed sequentially to [14C]- and [3H]leucine in association with stimulation by two 30 min pulses of the same secretagogue. Therefore, only [14C]hormone was available for release by the first stimulus, whereas both [14C]- and [3H]hormone were available for release by the second stimulus. Analysis was by specific immunoprecipitation. The first episode of stored [14C]rGH release exceeded the second episode of stored [14C]rGH release in response to each secretagogue. However, release of newly synthesized [3H]rGH in response to the second episode of stimulation exceeded the simultaneous release of stored [14C]rGH while matching or exceeding the [14C]rGH release that had occurred in the same experiment in response to the first episode of stimulation. Refilling both GH and PRL IRP stores drew primarily upon newly synthesized hormone, but with different secretagogue-specific patterns. These data confirm differential handling of new and stored rGH and rPRL within the pituitary. They are consistent with either (1) the enhanced shunting of newly synthesized hormone to IRPs within cells that are capable of compartmentalized intracellular hormone storage, or (2) the relatively complete discharge of a subset of somatotrophs and lactotrophs that are specialized to deliver pulsed hormone release, after which they are refilled by newly synthesized hormone. PMID- 2544409 TI - Epidermal growth factor increases intestinal calbindin-D9k and 1,25 dihydroxyvitamin D receptors in neonatal rats. AB - Epidermal growth factor (EGF) has been reported to increase intestinal calcium absorption in suckling rats. The mechanism of this effect is unknown, as are the roles of vitamin D-dependent and independent pathways. The present studies were undertaken to investigate the ability of EGF to accelerate the postnatal induction of the vitamin D-dependent intestinal calcium-binding protein, calbindin-D9k. Subcutaneous administration of EGF increased duodenal calbindin D9k in suckling rats by more than 100% (P less than 0.001). The effect of EGF was not seen in older weaned animals or when EGF was given to suckling rats by gavage. Administration of EGF simulated the changes of normal development. 1) It increased calbindin-D9k, and the effect was greater in proximal than distal duodenum. 2) EGF increased alkaline phosphatase activity to the same extent in proximal and distal duodenum. 3) EGF increased sucrase more markedly in distal than in proximal epithelium. Maximal and half-maximal effects of EGF on each of these proteins were observed at twice daily doses of 0.1 and 0.04 microgram/g BW, respectively. 4) EGF at the maximally effective dose produced a small (30%) but statistically significant (P less than 0.005) increase in serum 1,25 dihydroxyvitamin D. 5) Most importantly, EGF treatment resulted in a 2-fold increase in intestinal 1,25-dihydroxyvitamin D receptors (VDR) in the proximal segments of the small intestine (P less than 0.001). EGF effects on calbindin-D9k and VDR were specific for the intestine, as EGF did not change kidney calbindin D9k or kidney VDR. Thus, EGF was able to prematurely initiate a complex series of molecular changes that occur during normal development. The mechanism of EGF's action to stimulate calcium absorption appears to involve a maturation effect on the vitamin D-dependent pathway. PMID- 2544410 TI - Production of renin, angiotensin II, and aldosterone by adrenal explant cultures: response to potassium and converting enzyme inhibition. AB - The existence of renin in a number of extrarenal tissues has been well documented, but the physiological role of extrarenal renin remains unknown. To study the physiological role of adrenal renin, we developed a serum-free culture system for adrenal capsular/zona glomrulosa explants. Explants showed good viability in culture (greater than 80%), and demonstrated net production of aldosterone, angiotensin II, and prorenin. Aldosterone production was consistently stimulated by an increase in potassium (6 mM) in the culture medium. Both aldosterone and angiotensin II production could be attenuated by adding the angiotensin converting enzyme inhibitor lisinopril to the culture medium (0.1 mM). These data suggest that rat adrenal explants are capable of producing all of the components of a functional renin-angiotensin-aldosterone system and that these components can interact in response to physiological stimuli. These findings support the hypothesis that a local adrenal renin system may play a physiological role in the control of adrenal aldosterone production. PMID- 2544411 TI - The involvement of Leydig cells in the regulation of inhibin secretion by the testis. AB - The stimulation of Leydig cells by the administration of a single injection of 100 IU human CG (hCG) to adult male rats caused a significant biphasic stimulation of serum testosterone levels at 2 h and 3 days after injection. Serum immunoreactive (IR)-inhibin levels were elevated by 6 h and peaked at 24 h after hCG, then progressively declined thereafter. The removal of Leydig cells in vivo by the injection of the cytotoxic drug ethane dimethane sulfonate (EDS) causes a significant decrease in serum testosterone levels within 4 days, which is sustained 1 and 2 weeks after EDS. Serum IR-inhibin levels, however, rise significantly 2 and 4 weeks after injection of EDS. An injection of 100 IU hCG, 4 days after EDS (when no Leydig cells were present in vivo), failed to provoke an elevation of either testosterone or IR-inhibin levels in serum. But 2 or 4 weeks after administration of EDS, as a new population of Leydig cells develops in the interstitium, an injection of 100 IU hCG provokes a significant increase in serum testosterone and IR-inhibin levels. The possibility that the failure of IR inhibin levels to rise after EDS and hCG treatment could be due to changes in the seminiferous epithelium, caused by testosterone deprivation induced by Leydig cell destruction after EDS, was examined by administering high doses of testosterone known to maintain spermatogenesis. Under such conditions, hCG failed to induce a rise of IR-inhibin after EDS treatment had destroyed the Leydig cells. These data strongly support the concept that the Leydig cells are involved in the regulation of IR-inhibin secretion in vivo through factors other than testosterone. PMID- 2544412 TI - Different gamma-aminobutyric acid receptor subtypes are involved in the regulation of opiate-dependent and independent luteinizing hormone-releasing hormone secretion. AB - The neurotransmitter gamma-aminobutyric acid (GABA) appears to be involved in the control of gonadotropin secretion. These studies were conducted 1) to evaluate the effect of GABAergic drugs on in vitro LHRH secretion and 2) to characterize the role of different types of GABA receptors (the GABA-A and GABA-B subtypes) in these actions. Arcuate nuclei-median eminence fragments were incubated in vitro, and the release of LHRH, prostaglandin E2 (PGE2), arginine vasopressin, and oxytocin was measured by RIA. Both GABA and muscimol at different concentrations induced an increase in LHRH release, but did not affect the release of arginine vasopressin and oxytocin. This stimulatory effect was blocked by the specific GABA antagonist bicuculline, suggesting the involvement of GABA-A type receptors. Muscimol-stimulated LHRH release was not affected by the presence of phentolamine, suggesting that the stimulatory effect of GABA-A receptors on LHRH release is not mediated by interactions with the noradrenergic system. PGE2 has been shown to be a potent secretagogue of LHRH from the median eminence in vitro, and in this model the stimulatory effect of PGE2 was enhanced by muscimol. Baclofen, a specific GABA-B type receptor agonist, had no effect on basal LHRH release, but completely suppressed naloxone-stimulated LHRH and PGE2 secretion. The inhibitory effect of baclofen was blocked by the presence of 5-aminovalerate, a drug that has been shown to block the inhibitory effect of baclofen on NE release from noradrenergic terminals. This suggests the possibility that GABA-B receptors interacting with noradrenergic terminals may be responsible for the inhibitory effect of baclofen on naloxone stimulation. This study uncovered both stimulatory and inhibitory effects of GABA on LHRH release after activation of GABA-A or GABA-B receptors, respectively. Further, the data show possible relationships among the GABAergic, endogenous opiate peptide, and noradrenergic systems in the control of LHRH release from the hypothalamus. PMID- 2544413 TI - Characterization of anterior pituitary target cells for arginine vasopressin: including cells that store adrenocorticotropin, thyrotropin-beta, and both hormones. AB - Arginine vasopressin (AVP) stimulates the secretion of ACTH and TSH. Recently, affinity cytochemical techniques with a potent biotinylated analog of AVP were used to identify ACTH cells as one of the target cells. Counts showed that AVP bound cells were 10% of the population. However, if AVP bound all corticotropes and thyrotropes, one would expect AVP to bind at least 16% of the pituitary cells. Therefore, dual cytochemical labeling protocols were used to learn if thyrotropes also bound AVP (bio-AVP). Forty-eight percent of AVP target cells contained ACTH, and 42% contained TSH beta. The percentages of AVP-bound cells were increased to 12-13% of the total pituitary cells after 1-h pretreatment in 10 nM TRH or CRH. TRH and CRH together stimulated increases to 16% of the total cells. Analysis of dual labels showed that the additional AVP-bound cells stimulated by CRH or TRH stemmed from the corticotrope or thyrotrope populations, respectively. TRH stimulated an increase in the percentage of TSH cells that bound AVP from 55% to 75%. Similarly, CRH stimulated an increase in the percentage of ACTH cells that bound AVP from 61% to 79%. In addition, the populations of cells labeled for TSH beta or ACTH antigens increased by 30% after 1 h in unlabeled AVP, supporting its direct effect on these target cells. TRH stimulated a similar increment in TSH cells. CRH pretreatment had no effect on the percentages of cells labeled for TSH or ACTH. This could be the result of loss of ACTH stores needed to identify stimulated corticotropes. Finally, analysis of the total percentages of AVP-bound TSH beta or ACTH cells suggested an overlap in the population. This stimulated the application of dual labels for ACTH and TSH beta. In populations exposed to vehicle only, 1-2% of mixed pituitary cells stored both ACTH and TSH. This unique cell type also comprised 10% of a corticotrope population enriched by counterflow centrifugation. The percentage of ACTH-TSH cells in the mixed cell population was augmented to 4.8% after 1 h in AVP. It was not affected by exposure to either TRH or CRH (or both peptides). These studies demonstrate that AVP target cells include thyrotropes, corticotropes, and unique cells that store both ACTH and TSH. PMID- 2544414 TI - Interleukin-6 stimulates anterior pituitary hormone release in vitro. AB - Interleukin-6 (IL-6), a cytokine produced by inflammatory reactions, was found to stimulate PRL, GH and LH release from anterior pituitary cells at concentrations similar to those which affected lymphocyte mitogenesis. Perifused pituitary cells responded to IL-6 with prompt increases in hormone release that declined rapidly following cessation of exposure. Dopamine (DA) attenuated IL6-induced PRL release. In addition, IL-6 potentiated both GHRF- and TRH-induced hormone release without an affect on intracellular cAMP. These data demonstrate a new biological activity for IL-6 and provide evidence for immune system regulation of anterior pituitary hormone release. PMID- 2544415 TI - Sequential expression of high and low density lipoprotein receptors in differentiating fetal rat adrenocortical cells in primary culture. AB - Binding of fluorescent human high density lipoprotein-3 (DIL-HDL3) and fluorescent human low density lipoprotein (DIL-LDL) to differentiating fetal rat adrenocortical cells was studied in primary culture, a system in which addition of ACTH induces differentiation of zona glomerulosa cortical cells into zona fasciculata-type cells. Undifferentiated adrenocortical cells did not bind DIL HDL3 or DIL-LDL. Differentiation induced by ACTH was accompanied by intense binding of DIL-HDL3 and DIL-LDL to plasma membranes of adrenocortical cells. Binding of DIL-HDL3 and DIL-LDL was saturable and specific, and resulted in enhanced secretion of corticosterone and 18-hydroxydeoxycorticosterone, indicating that the binding sites for these lipoproteins represented functional receptors. Binding of DIL-HDL3 to the surface of adrenocortical cells was detected 6 h after the start of ACTH stimulation, whereas binding of DIL-LDL could not be detected until 12 h after ACTH stimulation was started. Thus, in the ACTH-induced differentiation of adrenocortical cells from zona glomerulosa-type into zona-fasciculata type, exogenous lipoprotein cholesterol is needed for enhanced growth and steroidogenesis, and an initial event in the differentiation of these cells is the sequential expression of specific receptors for HDL3 and LDL. PMID- 2544416 TI - Free fatty acids suppress growth hormone, but not luteinizing hormone, secretion in sheep. AB - An experiment was conducted to determine the effects of exogenously administered FFA on GH and LH secretion in sheep. Ovariectomized ewes received iv infusions of a mixture of FFA (166 mg/min; n = 5) or 0.9% saline (n = 4) for 10 h. Jugular blood was sampled every 15 min for 14 h, beginning 4 h before initiation of infusion. After 8 h of FFA or saline treatment, each ewe received a pituitary challenge of 10 micrograms GRF and 1 microgram GnRH, administered together as an iv bolus. Lipid infusion increased (P less than 0.01) serum FFA concentrations to levels characteristic of those in fasted sheep [23.0 +/- 0.8 mg/100 ml (mean +/- SE)]. Frequency of GH pulses (P less than 0.01) and the GH response to GRF (P less than 0.0001) were suppressed by FFA treatment. Mean serum GH concentrations increased gradually (P less than 0.01) during the 10-h infusion period in saline treated but not lipid-treated, ewes. This finding may reflect diurnal changes in somatotrope secretory activity that are blocked by FFA. Mean serum LH concentrations, LH pulse frequency and amplitude, and the LH secretory response to GnRH were unaffected by FFA or saline infusion. In agreement with previous work in sheep and other species, these results provide evidence for an inhibitory effect of FFA on GH release. The exact mechanism responsible for this action, however, remains to be elucidated. Finally, acutely elevated FFA levels do not appear to influence LH secretion in the ovariectomized ewe. PMID- 2544418 TI - Type 2 diabetes heredity and nutrient intake. A dietary history assessment in non obese normoglycaemic men. AB - The impact of type 2 diabetes heredity on nutrient intake was studied, by means of dietary histories, in 51 normoglycaemic, non-obese men, aged 54-59 years; 29 with familial aggregation of type 2 diabetes, and 22 with no such family history. The average daily intake of energy, macronutrients and minerals was almost identical in the two groups. Mean energy intake was approximately 2400 kcal/d, about 15 per cent of the energy deriving from protein, 35 per cent from fat, 45 per cent from carbohydrate and 5 per cent from alcohol. The average daily intake or dietary fibre was approximately 17 g, or 7 g/1000 kcal. Mean daily sodium and potassium intake, estimated from food sources, was about 3000 and 4000 mg, respectively. The findings provide no support for the existence of any relationship between type 2 diabetes heredity and dietary habits or nutrient intake. PMID- 2544417 TI - Nutritional status of infants aged 4 to 18 months on macrobiotic diets and matched omnivorous control infants: a population-based mixed-longitudinal study. I. Weaning pattern, energy and nutrient intake. AB - Information on food intake during weaning was collected as part of a mixed longitudinal study on the nutritional status and growth of the 1985 Dutch birth cohort of infants on macrobiotic diets (n = 53) and a matched control group on omnivorous diets (m = 57). Weighed food records over 3d, including breast-milk, were obtained on 49 macrobiotic and 57 control infants at 2-monthly intervals between the ages of 6 and 16 months. Intake of energy and nutrients was calculated using the Dutch food composition table which was supplemented by our own analyses of 50 macrobiotic foods. Ninety-six per cent of the macrobiotic infants and 74 per cent of the control infants had been breast-fed, but breast feeding continued longer in the macrobiotic group (13.6 vs 6.6 months, P less than 0.001). In the macrobiotic group, complementary feeding started at 4.8 months with water-based cereal porridges, followed later by vegetables, sesame seeds and pulses. Fruits were rarely given and products of animal origin were avoided. For all age groups combined, the intake of energy, protein, fat, calcium, riboflavin and vitamin B12 was significantly lower in the macrobiotic infants, whereas their intake of polysaccharides, fibre, iron and thiamin was higher than that of the control infants. The macrobiotic weaning diet tended to be bulky with a low energy density (2.4 kJ/g, controls: 3.4 kJ/g, P less than 0.05) and a high fibre content. Protein intake of the macrobiotic infants was only 80 per cent of the Dutch recommended daily intakes at the age of 6-8 months, and at 8 months, 69 per cent of this was derived from plant sources. Calcium intake was 280 mg/d; correction for calcium derived from hard tap-water raised the calcium intake to 308 mg in the macrobiotic age group of 14 months. The evidence of biochemical deficiencies of iron, riboflavin, vitamin B12, vitamin D and calcium is discussed. It is suggested that the macrobiotic diet should be supplemented with fat, fatty fish and dairy products. PMID- 2544419 TI - Mutagenicity of the human carcinogen treosulphan in Salmonella. AB - The human carcinogen treosulphan was mutagenic in Salmonella typhimurium TA100 and TA1535, as was dl-1,2:3,4-diepoxybutane (DEB), a proposed hydrolysis product of treosulphan. Another proposed hydrolysis product, methane- sulfonic acid, was not mutagenic in these strains. The pattern of the mutagenic responses at pH 6,7, and 8 to treosulphan and DEB suggests that DEB formation may be responsible for the mutagenicity of treosulphan. PMID- 2544421 TI - Calcium chelator Quin 2 prevents hydrogen-peroxide-induced DNA breakage and cytotoxicity. AB - Exposure of cultured Chinese hamster ovary (CHO) cells to hydrogen peroxide results in the production of extensive DNA breakage which can be prevented by the intracellular calcium chelator Quin 2. This effect occurs at Quin 2 AM concentrations as low as 0.1 microM and is maximal at 1 microM. Addition of the extracellular calcium chelator, EGTA, does not affect the level of DNA breakage generated by H2O2. Quin 2 also significantly reduces cellular toxicity caused by the oxidant. Experiments with spin-trapping techniques demonstrate that Quin 2 does not affect the formation of hydroxyl radicals generated by the action of Fe2+ on H2O2. Quin 2 at high concentrations, similar to those reached within the cell, actually enhanced generation of hydroxyl radical in the absence of other iron chelators under our experimental conditions. These results suggest that H2O2 or H2O2-derived radicals do not directly induce DNA strand breakage in intact mammalian cells; rather, these radicals may disturb intracellular Ca2+ homeostasis which results in secondary reactions ultimately leading to DNA strand breakage. In addition to strand breakage, membrane and protein oxidation probably contribute to the cytotoxic effect of H2O2. PMID- 2544420 TI - Age- and training-related changes in the collagen metabolism of rat skeletal muscle. AB - The effects of ageing and life-long endurance training on the collagen metabolism of skeletal muscle were evaluated in a longitudinal study. Wistar rats performed treadmill running 5 days a week for 2 years. The activities of collagen biosynthesis enzymes, prolyl-4-hydroxylase and galactosylhydroxylysyl glucosyltransferase, were highest in the muscles of the youngest animals, decreased up to the age of 2 months and from then on remained virtually unchanged. The enzyme activity in young animals was higher in the slow collagenous soleus muscle than in the rectus femoris muscle. The enzyme activity in the soleus muscle was higher for older trained rats than older untrained rats. The relative proportion of type I collagen increased and that of type III collagen decreased with age, suggesting a more marked contribution by type I collagen to the age-related accumulation of total muscular collagen. The results show that collagen biosynthesis decreases with maturation and that life-long endurance training maintains a higher level of biosynthesis in slow muscles. PMID- 2544422 TI - Ethanol-induced inhibition of leukotriene degradation by omega-oxidation. AB - omega-Oxidation of leukotrienes is a major pathway in the degradation and inactivation of these proinflammatory mediators. Ethanol inhibited this process in vivo and in vitro. In rat liver in vivo the catabolism of LTE4 to omega carboxylated leukotrienes was inhibited by 57% by an ethanol dose of 25 mmol/kg body mass administered intragastrically. The site of inhibition was the oxidation of omega-hydroxy-N-acetyl-LTE4 to omega-carboxy-N-acetyl-LTE4 resulting in an accumulation of omega-hydroxy-N-acetyl-LTE4 and of N-acetyl-LTE4. Analogous results were obtained for the oxidative degradation of LTB4 and omega-hydroxy LTB4 in rat hepatocyte suspensions. Ethanol, at a concentration of 12.5 mmol/l (0.07%; by vol.), caused 68% inhibition of the oxidation of omega-hydroxy-LTB4 by 50% in hepatocyte suspensions. The conversion of omega-hydroxy-LTB4 to omega carboxy-LTB4 by rat and human liver cytosol was inhibited by ethanol with half maximal concentrations of 100 mumols/l and 300 mumols/l, respectively. Our measurements indicate that direct interference by ethanol of the omega-oxidation of leukotrienes as well as an increased NADH/NAD+ ratio induced by ethanol led to the inhibition of leukotriene degradation. The impairment of leukotriene inactivation in the liver by ethanol may contribute to the development of the inflammatory reaction in acute alcoholic liver disease. PMID- 2544423 TI - Isolation and characterization of a rat liver alkaline phosphatase gene. A single gene with two promoters. AB - Structural analysis of 55 nearly full-length cDNA clones revealed heterogeneity in the 5'-untranslated regions of rat liver alkaline phosphatase mRNAs. The 5' extremities diverged into two totally unrelated sequence stretches at the position 88 nucleotides upstream of the initiation codon ATG. These two sequences, referred to as E1 and E2, were assigned on the genome about 36,000 base pairs (36 kbp) and 10 kbp upstream, respectively, of the exon coding for the 5'-most part of the common region. The gene consisted of 13 exons, including E1 and E2, and spanned about 56 kbp. The 11 exons (E3 to E13) following E1 and E2 were shared in common by the E1-type and the E2-type mRNAs. Analyses by S1 nuclease mapping and primer extension revealed the presence of two independent transcription-initiation sites specific to each of the E1 and E2 sequences. These results are interpreted as indicating a possible alternative usage of two leader exons, hence the presence of two independent promoters. Structural features of these putative promoters are described in the context of transcriptional fundamental and regulatory cis-elements. PMID- 2544424 TI - Spectroscopic characterization of the nickel and iron-sulphur clusters of hydrogenase from the purple photosynthetic bacterium Thiocapsa roseopersicina. 1. Electron spin resonance spectroscopy. AB - The thermostable hydrogenase from Thiocapsa roseopersicina was examined by low temperature ESR spectroscopy. Two types of signals were detected, from an oxidized iron-sulphur cluster and a nickel centre (Ni-A). In the oxidized protein additional signals were observed due to spin-spin interaction between the two paramagnetic centres. This interaction could be reversibly abolished by reduction to a redox potential below 105 mV. This implies that an additional redox centre is involved in the interaction, for which an Fe3+ ion is suggested. Reduction with hydrogen induced a second type of nickel ESR signal (Ni-C), corresponding to an intermediate redox state seen in other nickel hydrogenases. The Ni-C species was light-sensitive at cryogenic temperatures. At temperatures near to 4.2 K the Ni-C signal showed evidence of interaction with another paramagnetic centre, presumably a second iron-sulphur cluster. On reoxidation a signal due to a third Ni(III) species, Ni-B, increased in amplitude. These results establish that metal centres in the hydrogenase from T. roseopersicina are closely similar to those of the well-studied hydrogenase from Chromatium vinosum. PMID- 2544425 TI - Spectroscopic characterization of the nickel and iron-sulphur clusters of hydrogenase from the purple photosynthetic bacterium Thiocapsa roseopersicina. 2. Electron spin-echo spectroscopy. AB - Pulsed electron-spin-resonance techniques were applied to the hydrogenase of the purple photosynthetic bacterium Thiocapsa roseopersicina, an enzyme which contains nickel and iron-sulphur clusters but no flavin. The linear electric field effect profile of the spectrum in the region of g = 2.01 indicated that the strong ESR signal in the oxidized protein is due to a [3Fe-4S] cluster. The electron spin-echo envelope of this spectrum was modulated by hyperfine interactions with 1H and 14N nuclei, probably from the polypeptide chain. The ESR spectrum of this species shows a complex pattern arising from spin-spin interaction with another paramagnet. When the protein was partially reduced by ascorbate plus phenazine methosulphate, the complexity of the spectrum was abolished but the form of the electron spin-echo envelope modulation (ESEEM) pattern was unchanged. This indicates that the reversible disappearance of the spin-spin interaction pattern on partial reduction is not due to cluster interconversion to a [4Fe-4S] cluster. In the ESR spectrum of nickel(III), weak hyperfine interactions with 1H and 14N were also observed by ESEEM. The nature of the interacting nuclei is discussed. PMID- 2544426 TI - On the regulatory significance of inhibitors acting on non-equilibrium enzymes in the Calvin photosynthesis cycle. AB - Control analyses and kinetic model studies have been performed in order to obtain quantitative information on the regulatory significance of 12 experimentally well documented inhibitory interactions of Calvin cycle intermediates with the four non-equilibrium cycle enzymes. Evidence is presented to show that none of these interactions contributes significantly to the cycle flux control over the range of external orthophosphate concentrations where the reaction cycle shows close to optimal activity. Contrary to what has been generally supposed, the examined inhibitions appear to be of little interest for our understanding of the biological regulation of the Calvin photosynthesis cycle under conditions of light and carbon dioxide saturation. PMID- 2544427 TI - Insect immunity. Isolation of cDNA clones corresponding to diptericin, an inducible antibacterial peptide from Phormia terranovae (Diptera). Transcriptional profiles during immunization. AB - We have previously isolated and characterized a family of novel 8-kDa cationic antibacterial peptides synthesized by larvae of Phormia terranovae (Diptera) in response to various injuries. These molecules have been named diptericins. The peptide sequence of diptericin A was used to prepare oligonucleotides for screening cDNA libraries and we report in the present paper the isolation of several cDNA clones encoding diptericin. The analysis of the nucleotide sequences indicates that diptericin is synthesized as a prepeptide which is matured in two steps: (a) cleavage of a signal peptide and (b) amidation of the C-terminal residue. Interestingly, the 3' untranslated region of the mRNA contains a consensus sequence TTATTTAT which is also observed in the mRNA of another insect antibacterial peptide (attacin-related sarcotoxin IIA) and in mRNAs encoding proteins related to the inflammatory response in mammals. Our data illustrate that diptericins form a polymorphic family of immune peptides. The transcription of the diptericin genes is rapidly induced in the fat body after inoculation of bacteria, as evidenced by the transcriptional profile. PMID- 2544428 TI - Spectral and kinetic studies of phosphate and magnesium ion binding to yeast inorganic pyrophosphatase. AB - Inorganic pyrophosphatase must bind two phosphate molecules in order to catalyze pyrophosphate synthesis. In this report it is shown that Pi causes marked effect on the absorption spectrum of baker's yeast inorganic pyrophosphatase and this effect can be used to analyze Pi binding to this enzyme. A series of absorbance versus Pi concentration curves in the presence of 0.5-20 mM free Mg2+ were obtained at pH 7.2 and computer-fitted to 19 models. The dissociation constant of magnesium phosphate (8.5 +/- 0.4 mM) used in this analysis was measured with a Mg2+-sensitive electrode. The best model implies successive binding of two magnesium phosphate molecules or random-order binding of magnesium phosphate and free phosphate molecules. The first route predominates at physiological concentrations of Mg2+. The Pi-inhibition pattern of pyrophosphate hydrolysis confirmed that Pi adds to the active site and provided further evidence for the existence of an activating Pi-binding site. The possibility is raised that the pathways of pyrophosphate synthesis and hydrolysis by inorganic pyrophosphatase may differ in the sense that the binding of the fourth metal ion/subunit may facilitate the synthesis and inhibit the hydrolysis. PMID- 2544429 TI - Phase II studies of mitozolomide in melanoma, lung and ovarian cancer. AB - Seventy-seven patients were treated with oral mitozolomide to assess the activity of this drug in melanoma, lung and ovarian cancer. Partial responses were seen in five of 18 evaluable patients with small cell lung cancer (SCLC) and three of 20 with melanoma. No activity was apparent in non small cell lung or epithelial ovarian cancer. The major toxicity was myelosuppression which necessitated reduction in the initial dosage from 115 to 90 mg/m2. However, even at this dose level, unpredictable WHO grade 4 toxicity occurred in non-pretreated patients. Thrombocytopenia was more common than leucopenia and eight patients required platelet transfusion for spontaneous or tumour-related haemorrhage. Myelotoxicity was considered responsible for two deaths and was a significant contributory factor in a further three. Non-haematological toxicity was minor. Thus, despite demonstrable activity in SCLC and melanoma, unpredictable myelosuppression is likely to preclude further assessment in combination chemotherapy regimes in these tumours. PMID- 2544430 TI - Ischemia effect on the level of steroid hormone receptors in hepatocellular carcinoma and adjacent liver tissue. PMID- 2544431 TI - The effects of food and dose on the oral systemic availability of itraconazole in healthy subjects. AB - We have studied the influence of food and dose (50, 100, 200 mg) on the oral systemic availability of the broad spectrum antifungal itraconazole and the pharmacokinetics after repeated dosing of 100 mg in six healthy volunteers. The relative systemic availability of itraconazole capsules compared with solution averaged 39.8% in the fasting state but 102% in the post-prandial state. Food did not significantly affect the tmax of the capsules. Itraconazole AUC at single doses of 50, 100, and 200 mg had a ratio of 0.3:1:2.7, and the steady-state AUC (0-24) after 15 days of 100 mg was five times the single-dose AUC. These findings suggest non-linear itraconazole pharmacokinetics in the range of therapeutically used doses. Furthermore, capsules should be given shortly after a meal to ensure optimal oral systemic availability. PMID- 2544432 TI - Human T cell activation: differential response to anti-CD28 as compared to anti CD3 monoclonal antibodies. AB - Monoclonal antibodies (mAb) against CD3 or CD28 in conjunction with the tumor promoter phorbol 12-myristate 13-acetate (PMA) induce interleukin 2 receptor (IL2R) expression, IL2 production and proliferation in resting T cells. Recent studies indicate that these two pathways are biochemically distinct. In this study T cell activation induced by PMA and anti-CD28 mAb 9.3 is compared to the effects of PMA plus anti-CD3 mAb (T3-II and 235) in the presence or absence of cyclosporin A (CsA), dibutyryladenosine 3':5' cyclic monophosphate (db-cAMP) or cholera toxin (CT). Proliferation of T cells stimulated with PMA plus mAb 9.3 is resistant to the inhibitory effects of CsA, db-cAMP and CT. Only at the highest dose did CsA have any effect on PMA plus mAb 9.3-induced T cell proliferation. Conversely, CsA, db-cAMP and CT inhibit PMA plus T3-II-induced T cell proliferation. mRNA analysis further demonstrates the similarities and the differences between the CD28 and CD3 activation pathways. Recently, T3-II was reported to induce tumor necrosis factor (TNF) and lymphotoxin (LT) mRNA synthesis in PMA-treated T cells. In this study mAb 9.3 is shown to substitute for T3-II in the induction of TNF and mRNA. However, the production of TNF and LT mRNA in PMA plus mAb 9.3-treated T cells is greater than that seen in PMA plus T3 II-treated cells. mRNA synthesis included by PMA plus T3-II is blocked by CsA. mRNA production in T cells activated with PMA plus mAb 9.3 is resistant to CsA. Similar results are noted with IL2 and IL2R mRNA. Flow cytometric analysis of the IL2R confirms the mRNA data. CsA blocks the T3-II-induced potentiation of PMA induced IL2R expression but not the mAb 9.3-induced potentiation. This differential inhibitory effect of CsA on IL2R expression is also seen with db cAMP and CT. We examined the effects of these two pathways on the expression of the early activation antigen EA 1 and cytoplasmic free calcium. Recently, we have shown anti-CD3 mAb potentiate EA 1 expression induced by 1,2-sn dioctanoylglycerol and this potentiation is calcium dependent. dp-cAMP blocks T3 II- and 235-induced potentiation of EA1 expression and inhibits the T3-II- and 235-mediated rise in intracellular free calcium [( Ca2+]i). Conversely, 9.3 does not potentiate EA 1 expression or induce a rise in [Ca2+]i. These results provide further evidence that the CD28 and CD3 activation pathways utilize distinct signal transduction pathways. PMID- 2544434 TI - Collagen receptor on T lymphocytes and the control of lymphocyte motility. AB - Human lymphocytes, freshly isolated from blood, were allowed to settle on surfaces coated by collagen type 1, fibronectin, laminin, IgG or albumin at different concentrations. In separate cultures the lymphocytes were also exposed to these proteins in soluble form. The lymphocytes, predominantly T cells, attached to two-dimensional collagen substrata both in the presence and absence of serum but did not adhere or adhered poorly to substrata coated with fibronectin, laminin, IgG and albumin. In contrast, T blasts induced in a mixed lymphocyte culture adhered to fibronectin-coated substratum. During contact with substratum-bound collagen for a 24-h period, 47 +/- 15% of the freshly purified lymphocytes from separate individuals developed motile behavior whereas 16 +/- 4% of the cells became motile on fibronectin. Gelatin (denatured collagen) also mediated attachment of lymphocytes to surfaces but only at comparatively high concentrations (40 mg/ml). Collagen and gelatin in solution also caused agglutination and motility of the vast majority of freshly isolated T lymphocytes whereas fibronectin and other proteins, when presented in soluble form, did not. Cell agglutination was maximal at moderate (10 or 20 mg/ml) and cell motility at low gelatin concentrations (1 to 10 mg/ml). High gelatin concentrations (20 and 40 mg/ml) did not induce motile behavior. Cytochalasin B augmented the proportion of adherent cells on gelatin-coated substrata. In the presence of cytochalasin B gelatin mediated substrate-adhesion at concentrations below those which normally induced adhesion indicating that motile behavior counteracted persistent lymphocyte adhesion to the substratum. Noteworthy, gelatin/collagen is unique among ligands (e.g. plasma fibronectin and other serum proteins) in its capacity to induce motility in the vast majority of resting lymphocytes freshly isolated from blood within a relatively short period. Taken together these results indicate that circulating lymphocytes have a collagen/gelatin-binding plasma membrane component. Cross-linking of this component is a likely explanation for the selective inducing effect of gelatin and collagen on lymphocyte motility. The present results showed that the lymphocyte plasma membrane contains collagen binding components with a relative molecular mass of 130 and 55 kDa. The 55-kDa component also reacted with an anti-fibronectin antibody. Thus, interactions with the extracellular matrix may control lymphocyte locomotor capacity. PMID- 2544433 TI - Interleukin 1 and tumor necrosis factor enhance transcription from the SV40 early promoter in a T cell line. AB - Interleukin 1 (IL 1) and tumor necrosis factor (TNF) increase the expression of a number of T lymphocyte-specific genes in the T-cell hybrid PC60. We show here that human IL 1 alpha and 1 beta, mouse IL 1 alpha and mouse TNF-alpha strongly enhance expression of reporter genes transcribed from the SV40 early promoter in this cell line. We found that IL 1 and TNF each induce up to a 30-fold increase in the rate of transcription, resulting in a proportional increase of mRNA and protein levels. Induction with IL 1 and TNF is detectable after 1 h and reaches a plateau after about 15 h. Removal of these factors from the culture medium results in complete reversion. Induction with IL 1, but not with TNF, can be inhibited with an inhibitor of IL 1 binding. The effects of both factors are not impaired by the protein synthesis inhibitor cycloheximide, suggesting that they stimulate expression from the SV40 early promoter by activation of preexisting transcription factors. PMID- 2544435 TI - Corticotropin-releasing factor (CRF) stimulation test in normal subjects and patients with Cushing's syndrome. AB - The hormonal responses to human corticotropin-releasing factor (hCRF) were investigated in 6 normal subjects, 13 patients with Cushing's disease (8 with diffuse bilateral hyperplasia and 5 with nodular hyperplasia) and one patient with Cushing's syndrome due to an adrenal adenoma. hCRF (100 micrograms i.v.) was a potent stimulant of ACTH and cortisol in normal subjects. Patients with Cushing's disease due to diffuse hyperplasia showed variable ACTH and cortisol responses to hCRF. In both normal subjects and in patients hCRF consistently stimulated serum aldosterone levels. Patients with nodular hyperplasia had extremely suppressed plasma ACTH levels and no responses of ACTH, cortisol and aldosterone to hCRF like in the patient with adrenal adenoma. Our results suggest that hCRF-stimulation test may be a useful tool for differentiating pituitary and adrenal forms of Cushing's syndrome. PMID- 2544436 TI - Specific radioimmunoassay of alpha-melanocyte-stimulating hormone in rat plasma. AB - A sensitive, specific and reproducible radioimmunoassay was developed for the measurement of alpha-melanocyte-stimulating hormone (alpha-MSH) in the blood plasma of rat. The assay method is based on a sensitive antiserum raised against alpha-MSH in rabbit. The serum is highly specific to alpha-MSH; a HPLC study of an extract of rat plasma showed that the immunoreactivity was given by alpha-MSH. The basal level of alpha-MSH, measured after a simple extraction with ethanol, was found to be 168.3 +/- 16.3 pg/ml (mean +/- SEM). Ether and lysine-vasopressin appeared to be potent stimuli for the peripheral release of alpha-MSH. PMID- 2544437 TI - Changes of dipeptidyl peptidase (DP IV) activity in the T lymphocytes of rats following administration of ACTH, dexamethasone and opiates. AB - The aim of the work was to study the effect of glucocorticoids, opiates and stressful stimuli on dipeptidyl peptidase IV (DP IV, EC 3.4.14.5) activity of T lymphocytes prepared from the thymus of intact and adrenalectomized rats. Four week old male rats of Wistar strain were used. The in vivo administration of ACTH, dexamethasone and morphine treatment resulted in an increase of DP IV activity in the cell suspension. In adrenalectomized rats ACTH treatment failed to modify the enzyme activity, however, pain or emotional stress resulted in an elevated DP IV activity. Morphine and D-Met2-Pro5-enkephalinamide resulted in a dose dependent activation of DP IV in T cells, an effect which could be modified by naloxone pretreatment. Our findings show that DP IV mechanisms in T cells are highly sensitive to exogenous and endogenous steroids, opiates and biologically active substances released in response to stress in rats. PMID- 2544439 TI - The function of cellular transcobalamin II in cultured human cells. AB - The known function of human transcobalamin II (TC II) is to transport cobalamin (Cbl) in the circulation to tissue receptors for TC II-Cbl. Several types of human cells synthesize apo (unsaturated) TC II and the present study was conducted in order to evaluate possible functions of this endogenous TC II. The approach consisted of a correlation between the abilities of cultured cells to produce apo TC II and to internalized Cbl when presented in the free form. The amount of apo TC II produced by six lines of cultured human cells ranged from abundant to nil. The amount of free Cbl internalized by these cells correlated directly with the capacity to produce apo TC II. The interactions between endogenous TC II and free Cbl took place either at the cell surface or in the medium surrounding the cell. It was also shown that cells in culture contain free Cbl and release free Cbl into the surrounding medium. Thus it was concluded that the apo TC II produced by human cells remains intact to interact with free Cbl and to participate in the cellular metabolism of Cbl. PMID- 2544438 TI - Mechanism of fibronectin-mediated cell migration: dependence or independence of cell migration susceptibility on RGDS-directed receptor (integrin). AB - Cell migration on fibronectin (FN)-coated substrata was studied using 10 cell lines, of which only 2 showed clear enhancement and 1 showed marginal enhancement of cell migration. The migration of the other 7 cell lines was not affected on FN coated substrata, although they all showed FN-dependent cell adhesion. The migration-enhancing activity of FN was found in the fragment including the cell adhesion and Hep-2 domains, but not other domains (Hep-1/Fib-1, Gel, Fib-2). No difference in the migration-enhancing effect was seen among FNs from plasma, fibroblasts, or transformed cells. FN-dependent cell migration was inhibited by polyclonal antibodies directed to the C-terminal half region including the cell binding domain, but not by antibodies directed to five other domains. Since these results indicated that FN-mediated cell migration could be controlled by the cell adhesion domain of FN and its receptor, studies were then focused on the effect of antibodies directed to receptors for FN and collagen, and on the effect of tetrapeptide sequences recognized by these receptors. It was found that (i) cell migration on FN-coated surfaces was specifically inhibited by anti-FN receptor antibody P1F8 but not by anticollagen receptor antibody P1H5; (ii) the migration was strongly inhibited by Arg-Gly-Asp-Ser but not by other oligopeptide sequences. However, the majority of those cell lines not susceptible to FN dependent cell migration were characterized by having FN receptors and the ability to adhere on FN-coated matrix. Based on these findings, it was concluded that FN-dependent cell migration shares the same recognition mechanism as FN dependent cell adhesion, but that the majority of cell lines not exhibiting FN dependent migration still show FN-dependent cell adhesion and express the FN receptor (integrin); i.e., cell migration and adhesion involve the same receptor and the same FN loci, but migration is controlled by still-unidentified cellular factors which determine the susceptibility of the cell to the dynamic function of the FN receptor (integrin) unit. PMID- 2544440 TI - Growth factor requirements of normal and polyomavirus middle T gene transformed REF52 cells in serum-free medium: indications of a reduced vasopressin requirement and its relationship to the control of phosphatidylinositol metabolism. AB - The growth factor requirement of normal and polyomavirus middle T gene transformed REF52 cells was studied in serum-free medium in an attempt to elucidate the possible linkage between an altered growth factor requirement and one or more altered physiological properties of the transformed cells. For optimal growth, REF52 cells required vasopressin, epidermal growth factor (EGF), high-density lipoprotein (HDL), hydrocortisone, insulin, transferrin, and fibronectin. Deletion of vasopressin or hydrocortisone from the medium resulted in a 50 to 60% reduction in cell growth and the deletion of HDL, transferrin, or the combination of EGF and vasopressin led to an 80 to 90% growth retardation. The same medium supported the growth of the transformed variant (PyMLV-REF52) at a rate comparable to that of 10% serum, and deletion of hydrocortisone, vasopressin, or the combination of EGF and vasopressin had virtually no effect on PyMLV-REF52 cell growth. In vasopressin-deleted medium, vasopressin elicited a rapid increase of intracellular inositol phosphate levels in REF52 cells and the control of phosphoinositide turnover was strictly regulated. In contrast, both cell proliferation and intracellular inositol phosphate levels of PyMLV-REF52 cells were not affected by vasopressin treatment under identical culture conditions, and control of phosphoinositide metabolism was lost. Thus, a correlation may exist between the trigger of a mitogenic signal and the stimulation of the phosphoinositol pathway by vasopressin in REF52 cells and this relationship was disrupted in PyMLV-REF52 cells. PMID- 2544442 TI - Epidemiology of parainfluenza virus type 3 in England and Wales over a ten-year period. AB - We have analysed data on respiratory syncytial (RS) and parainfluenza type 3 (PF3) viruses reported to the Communicable Disease Surveillance Centre, London, over the period 1978-87. These confirm the annual winter epidemic of RS virus and show that, in England and Wales, PF3 is a summer infection with regular yearly epidemics. PMID- 2544441 TI - Evidence for a new rotavirus subgroup in India. AB - Monoclonal antibodies specific for rotavirus subgroup 1 (SG1) and subgroup 2 (SG2) were used to analyse by enzyme immunoassay (EIA) the subgroups of human rotavirus isolates obtained from three different parts of India during the period September 1985 to July 1987. We identified one isolate which failed to react with either SG1 or SG2 specific monoclonal antibodies, although it reacted well with a monoclonal antibody specific for group A rotaviruses. This finding suggests that it belongs to a new rotavirus subgroup. Further, another isolate was found to belong to SG1 although it had a 'long' electropherotype. PMID- 2544443 TI - Immunopharmacological analysis of endotoxin-induced uveitis in the rat. AB - Footpad injection of endotoxin causes exclusive ocular inflammation in the rat. In order to clarify its physiopathologic mechanism, we studied the effect of different treatments on endotoxin-induced uveitis (EIU). Salmonella endotoxin was injected into the footpads of Lewis rats. 18 hr later, inflammation was assessed by evaluating proteins and cells in the anterior chamber; arachidonic acid (AA) metabolites, prostaglandin E2 and leukotriene B4, as well as substance P were measured by radioimmunoassay, and Ia-(MHC class II)-antigen expression in ciliary body was assessed by immunohistochemistry. The effect of inhibitors of phospholipase A2 (EPC), of lipoxygenase (azelastine) and of cyclo-oxygenase (diclofenac), as well as dexamethasone, cyclosporine (CsA) and anti-Ia antibody, were evaluated on these parameters. Phospholipase A2 inhibitor EPC and dexamethasone were most effective on inflammation: they also reduced AA metabolites very effectively and prevented Ia-expression. Lipoxygenase and cyclo oxygenase inhibitors were partially effective on inflammation and on AA metabolites but failed to prevent Ia-expression. Immunosuppressive treatments (CsA and anti-Ia-antibody) also reduced inflammation. Our findings suggest that inflammation mediators initiate inflammation in EIU. Ia-Ag-expression is secondarily produced by mediators leading to additional inflammation due to immune mediated mechanisms. PMID- 2544444 TI - The inotropic effect of endothelin-1 on rat atria involves hydrolysis of phosphatidylinositol. AB - Endothelin-1 induces a positive inotropic response in isolated left atria of the rat with an IC50 value of 20 nM. The contractile effect of endothelin is larger than that of other inotropic hormones such as phenylephrine and epinephrine and smaller than that of Bay K8644. In the spontaneously active right atria, endothelin induces a positive inotropic effect with no chronotropic effect. Endothelin does not modify intracellular levels of cAMP under basal conditions or after stimulation with isoproterenol but stimulates the formation of inositol phosphates. Mobilization of inositol phospholipids is observed in the same range of concentrations as for the contractile action of endothelin. The contractile action of endothelin is not mediated by protein kinase C. It is antagonized by blockers of L-type Ca2+ channels, low external Ca2+ concentrations and drugs such as caffeine and ryanodine that interfere with Ca2+ release by the sarcoplasmic reticulum. PMID- 2544445 TI - Cytochrome o (bo) is a proton pump in Paracoccus denitrificans and Escherichia coli. AB - Spheroplasts from aerobically grown wild-type Paracoccus denitrificans cells respire with succinate despite specific inhibition of the cytochrome bc1 complex by myxothiazol. Coupled to this activity, which involves only b-type cytochromes, there is translocation of 1.5-1.9 h+/e- across the cytoplasmic membrane. Similar H+ translocation ratios are observed during oxidation of ubiquinol in spheroplasts from aerobically grown mutants of Paracoccus lacking cytochrome c oxidase, or deficient in cytochrome c, as well as in a strain of E. coli from which cytochrome d was deleted. These observations show that the cytochrome o complex is a proton pump much like cytochrome aa3 to which it is structurally related. PMID- 2544446 TI - Synthesis of an affinity ligand ('UPHIT') for in vivo acylation of the kappa opioid receptor. AB - The isothiocyanate analog (1S,2S-trans-2-isothiocyanato-4,5-dichloro-N- methyl-N [2-(1-pyrrolidinyl)cyclohexyl]benzeneacetamide, 3a) of the highly selective kappa opioid receptor agonist, U50,488, was prepared as a potential site-directed affinity ligand for acylation of kappa-opioid receptors in vivo. The isothiocyanate (3a) which we have designated UPHIT and its enantiomer (3b) were synthesized in 3 steps starting from optically pure (1S,2S)-(+)-trans-2 pyrrolidinyl-N-methyl-cyclohexylamine (4a) and its enantiomer (4b), respectively, thus defining their absolute stereochemistry. Binding in vitro of the 1S,2S enantiomer 3a to kappa receptors labelled by [3H]U69,593 was shown to occur with an IC50 value of 25.92 +/- 0.36 nM, whereas 827.42 +/- 5.88 and 115.10 +/- 1.23 nM were obtained for the IC50 value of the 1R,2R enantiomer (3b) and (+/-)-3 respectively. Intracerebroventricular (ICV) injection of 100 micrograms of (+/-) 3 into guinea-pig brain followed by analysis of remaining kappa-binding sites 24 h later revealed that (+/-)-3 depleted 98% of the kappa receptors that bind [3H]U69,593 and 40% of those that bind [3H]bremazocine. These preliminary data suggest exciting uses for these compounds in furthering our knowledge of the kappa-opioid receptor. PMID- 2544447 TI - Biphasic increase in intracellular calcium induced by platelet-activating factor in macrophages. AB - In single mouse macrophages stimulated by platelet-activating factor (PAF), the intracellular calcium concentration (Cai) monitored with fura-2 at room temperature presents a biphasic increase, including a transient and a more sustained component. After pulse administration of PAF, the first phase lasts for a few seconds and reaches a peak value of 0.5-1 microM Ca2+ at high PAF concentration. The amplitude of this peak is independent of extracellular Ca2+ concentration, suggesting that the initial Ca2+ transient is due to the release of Ca2+ from intracellular stores. The second phase of the response lasts for several minutes; its maximum amplitude is reached 1-2 min after the brief initial PAF stimulation. This phase, suppressed in zero external Ca2+ and increased in 10 mM Ca2+, is probably due to influx of Ca2+ through the plasma membrane. This secondary Ca2+ increase is blocked by 10-50 microM lanthanum. At low PAF concentration, the initial Ca2+ transient is not followed by a second phase, showing that the initial rises of Ca2+ and of its activator (presumably inositol trisphosphate) are not sufficient to trigger the second phase of Ca2+ increase. PMID- 2544448 TI - Forskolin stimulates pinealocyte cGMP accumulation. Dramatic potentiation by an alpha 1-adrenergic----[Ca2+]i mechanism involving protein kinase C. AB - The effect of forskolin on cGMP regulation was investigated using dispersed rat pinealocytes. Forskolin stimulated cGMP accumulation in a concentration-dependent manner; this response was strongly potentiated by an alpha 1-adrenergic--- [Ca2+]i mechanism involving protein kinase C. These findings provide further evidence that activation of two receptor-regulated signal transduction mechanisms may be commonly required for maximal stimulation of cGMP accumulation, and establish a new experimental approach to the study of cGMP regulation. PMID- 2544449 TI - Magainin 2 amide and analogues. Antimicrobial activity, membrane depolarization and susceptibility to proteolysis. AB - We compared the abilities of synthetic magainin 2 amide and its analogues to inhibit the growth of Escherichia coli and to cause membrane depolarization in E. coli cells and cytochrome oxidase liposomes. The analogue, magainin A, was about 40-times more active than magainin 2 amide in inhibiting the growth of E. coli and had a much more sustained effect on the membrane potential. In the liposomal system, however, there was only approx. 20% difference between these two peptides in the reduction of membrane potential and uncoupling of respiration. Studies with pronase digestion suggested that the difference in potency may be due to differential susceptibility to proteolysis in the presence of membranes. PMID- 2544450 TI - Phospholipase C and phospholipase A2 are involved in the antiviral activity of human interferon-alpha. AB - Treatment of human amniotic cells (UAC) with human interferon-alpha (Hu-IFN alpha) or phorbol myristate acetate (PMA) resulted in translocation of protein kinase C (PK-C) activity from the cytosol fraction to that of the membranes. Analysis of 32P incorporation into phospholipid fractions and studies of alterations in fatty acid content for the major phospholipids of IFN-treated cells suggest that phospholipases C and A2 are activated by Hu-IFN alpha. Addition of neomycin (an inhibitor of phospholipase C), as well as mepacrine (an inhibitor of phospholipase A2) to IFN-treated cells inhibited the antiviral activity of Hu-IFN alpha in the vesicular stomatitis virus (VSV)-UAC system used. These observations indicate that (i) activation of PK-C and (ii) diacylglycerol formation, arachidonic acid and/or lysophosphatidylcholine release are important steps in the mechanism of action of IFN. PMID- 2544451 TI - Neuropeptide Y inhibits vasoactive intestinal peptide- and dopamine-induced cyclic AMP formation in human Ewing's sarcoma WE-68 cells. AB - Neuropeptide Y (NPY) regulation of intracellular cyclic AMP accumulation was studied in human Ewing's sarcoma cell line, WE-68. NPY inhibited vasoactive intestinal peptide (VIP)- and dopamine-stimulated but not basal cyclic AMP formation. The peptide effect was rapid (less than 2 min), concentration dependent with a half-maximal effective concentration (EC50) of 8 nM NPY, and maximal inhibition reaching 60-70% with 100 nM NPY. Prior exposure of WE-68 cells to pertussis toxin completely abolished the inhibitory action of NPY. It is concluded that NPY attenuates agonist-stimulated cyclic AMP formation in Ewing's sarcoma WE-68 cells, and may do so via the inhibitory guanine nucleotide regulatory protein of adenylate cyclase. PMID- 2544452 TI - Expression of high- and low-affinity epidermal growth factor receptors in human hepatoma cell lines. AB - Data are presented from a comparative research on expression of epidermal growth factor (EGF) receptors and response to EGF of six independently established cell lines derived from human hepatoma. These lines differ in terms of the degree of differentiation, presence of hepatitis B virus (HBV) DNA copies in integrated form and expression of HBV genes. Our results indicate differential expression of membrane EGF receptors and differential response to EGF under serum- and hormone free culture conditions. Furthermore, a significant difference in affinity could be detected between EGF receptors of the two highly dedifferentiated cell lines (HA22T/VGH and Li7A) whose replication is inhibited by EGF concentrations capable of stimulating more differentiated phenotypes. PMID- 2544453 TI - Stimulation of phosphoinositide hydrolysis via class I antigen-specific recognition in murine cardiac tissue. AB - Induction of polyphosphoinositide hydrolysis in cardiac tissue by specific recognition of class I histocompatibility antigens was assayed. C3H (H-2k) mice auricles were labelled with myo-[3H]inositol precursor and inositol phosphate production in the presence or absence of anti-class I k products was measured. Anti-class I, but not anti-class II products specifically increased phosphoinositide turnover. This increment was partially blocked by muscarinic cholinergic and alpha-adrenergic blockers and even more so by the phospholipase C inhibitor NCDC. Alloantibodies specifically directed against class I antigens could then exert stimulation of phospholipase C-mediated phosphoinositide hydrolysis through the interaction with muscarinic cholinergic and/or alpha adrenergic receptors. The induction of intracellular second messengers by class I antigens and hormone-receptor interactions is discussed. PMID- 2544454 TI - Two components of type III protein kinase C with different substrate specificities and a phospholipid-dependent but Ca2+-inhibited protein kinase in rat brain. AB - The activities of rat brain protein kinase C isoenzymic fractions separated by hydroxyapatite chromatography were measured with histone H1 or the oligopeptide Ala-Ala-Ala-Ser-Phe-Lys-Ala-Lys-Lys-amide as substrates. The oligopeptide was a better substrate than histone H1 for nearly all of the protein kinase C fractions. Two subfractions of type III isoenzyme were resolved (IIIa and IIIb); type IIIb was characterized by a very low histone kinase activity compared to its peptide kinase activity. In some brain extracts a phospholipid-dependent but Ca2+ inhibited protein kinase was also observed which was eluted from the hydroxyapatite column between type II and III isoenzymes of protein kinase C. PMID- 2544455 TI - Activation of matrix metalloproteinase 3 (stromelysin) and matrix metalloproteinase 2 ('gelatinase') by human neutrophil elastase and cathepsin G. AB - The ability of human neutrophil elastase and cathepsin G to activate matrix metalloproteinase 3 (MMP-3 = stromelysin) and MMP-2 ('gelatinase') purified from human rheumatoid synovial fibroblasts in culture was examined. The zymogen of MMP 3 (proMMP-3) was activated to full activity with elastase and cathepsin G by limited proteolysis of the molecule into two active forms of Mr approximately 45,000 and Mr approximately 25,000. In contrast, proMMP-2 was not activated at all by these neutrophil serine proteinases, although it was degraded into small fragments. These data suggest that neutrophil elastase and cathepsin G may play an important role in the activation of proMMP-3 in vivo in various inflammatory conditions, but proMMP-2 may be activated in different ways. PMID- 2544456 TI - Calcitonin gene-related peptide-1 (CGRP-1) is a potent regulator of glycogen metabolism in rat skeletal muscle. AB - We investigated the effects of CGRP on glucose metabolism in intact rat skeletal muscle preparations that are largely composed of either type I (soleus) or II fibres (e.g. extensor digitorum longus (EDL) or epitrochlearis muscles). CGRP-1 inhibited insulin-stimulated glycogen synthesis in both soleus and EDL muscle preparations. Rat CGRP-1 was a potent stimulator of glycogenolysis only in muscles composed of type II fibres, which depend on high rates of glycogenolysis to produce high power outputs. These results may provide the basis for understanding how CGRP regulate glycogenolysis in type II fibres in vivo. PMID- 2544457 TI - Bacterial Na+ energetics. AB - Novel observations related to the Na+-linked energy transduction in bacterial membranes are considered. It is concluded that besides the well-known systems based on the circulation of protons, there are those based on the circulation of Na+. In some cases, H+ and Na+ cycles co-exist in one and the same membrane. Representatives of the 'sodium world', i.e. cells possessing primary Na+ pumps (delta mu Na generators and consumers) are found in many genera of bacteria. Among the delta mu Na generators, one should mention Na+-NADH-quinone reductase and Na+-terminal oxidase of the respiratory chain, Na+-decarboxylases and Na+ ATPases. For delta mu Na consumers, there are Na+-ATP-synthases, Na+-metabolite symporters and Na+ motors. Sometimes, one and the same enzyme can transport H+ or, alternatively, Na+. For instance, an Na+-ATP-synthase of the F0F1 type translocates H+ when Na+ is absent. Employment of the Na+ cycle, apart from or instead of the H+ cycle, increases the resistance of bacteria to alkaline or protonophore-containing media and, apparently, to some other unfavourable conditions. PMID- 2544458 TI - Maturation-dependent changes of the rabbit reticulocyte energy metabolism. AB - Rabbit reticulocytes were separated into four fractions of different maturity in order to investigate the changes of cellular respiration and glycolysis, adenine nucleotides, 2,3-biphosphoglycerate (2,3-BPG) as well as cyclic AMP level during the transition from the youngest to the most mature reticulocytes. A significant reduction of total oxygen consumption, mainly due to depression of coupled respiration was found. The decline of respiration was accompanied by a 2-fold increase of the rate of aerobic glycolysis indicating a reduced Pasteur effect during maturation. A decline of ATP and an increase of ADP concentration was found. The oxygen-delivery capacity of the red cells increased by about 26% caused by an increase of the 2,3-BPG level of about 2 mmol/l cells. Cyclic AMP level in the fraction of youngest reticulocytes was about 60-fold higher than that in mature rabbit erythrocytes. The biggest decline of cyclic AMP was registered during the transition from youngest to the intermediate stage of maturity. PMID- 2544459 TI - Mitochondrial F0F1 H+-ATP synthase. Characterization of F0 components involved in H+ translocation. AB - The membrane F0 sector of mitochondrial ATP synthase complex was rapidly isolated by direct extraction with CHAPS from F1-depleted submitochondrial particles. The preparation thus obtained is stable and can be reconstituted in artificial phospholipid membranes to result in oligomycin-sensitive proton conduction, or recombined with purified F1 to give the oligomycin-sensitive F0F1-ATPase complex. The F0 preparation and constituent polypeptides were characterized by SDS polyacrylamide gel electrophoresis and immunoblot analysis. The functional role of F0 polypeptides was examined by means of trypsin digestion and reconstitution studies. It is shown that, in addition to the 8 kDa DCCD-binding protein, the nuclear encoded protein [(1987) J. Mol. Biol. 197, 89-100], characterized as an intrinsic component of F0 (F0I, PVP protein [(1988) FEBS Lett. 237,9-14]) [corrected] is involved in H+ translocation and the sensitivity of this process to the F0 inhibitors, DCCD and oligomycin. PMID- 2544460 TI - The mechanism of proton translocation in respiration and photosynthesis. PMID- 2544461 TI - Structure of the alpha 1 subunit of horse Na,K-ATPase gene. AB - Genomic DNA for Na,K-ATPase alpha 1 subunit was obtained from libraries of horse kidney genomic DNA in Charon 4A and in EMBL3 bacteriophages by screening with the full sized cDNA probe of the alpha 1 subunit of rat Na,K-ATPase as probe. The gene spans 30 kb and consists of 23 exons and 22 intervening sequences. Intron exon boundaries were analyzed. The protein-coding nucleotide sequence encodes 1016 amino acids with an Mr of 112,264. The putative amino acid sequence of horse alpha 1 is 96-97% homologous to those of other mammalian species. PMID- 2544463 TI - Small carcinomas of the pancreas with associated chronic pancreatitis: two case reports. AB - Two cases of pancreatitis associated with papillary atypical hyperplastic lesions and micro-invasive cancers are reported. They are interesting from an etiopathogenic and diagnostic point of view by again illustrating the problem of transmission between hyperplastic lesions and cancer, together with the difficulties encountered in asserting the certainty of diagnosis. In each observation the association of pancreatic lesions, of all stages of hyperplasia, and of cancer can be observed. This suggests that transmission occurs between those different lesions. In such cases diagnosis of benignity or malignancy is always difficult to assert. No clinical, biological, morphological or even cytological criterion allows one to make a ruling. For this reason excision is recommended by the authors whenever the etiology of pancreatitis remains obscure and especially in the presence of important dystrophic lesions. PMID- 2544462 TI - Clinical and pathological features of breast cancers in Rivers State, Nigeria. AB - In this study, the clinico-pathological features of breast cancers seen in Rivers State of Nigeria were retrospectively analysed. Nine out of 67 patients (13%) were younger than 30 years of age. Only 13% of 46 patients presented within one month of the onset of symptoms. The mean duration of symptoms varied from 3.7 months in patients under 30 years of age to 13.3 months in those in the 50-59 years age range. Most patients, 40 out of 51, presented with axillary lymph node metastases, the incidence of which varied directly with both the duration of symptoms and age. It is suggested that the rise in the incidence of axillary lymph node metastasis with age is determined by increase in the duration of symptoms. These results highlight the need to intensify cancer education in the population. The presenting complaints and the distribution of the various histological types did not differ from other reports. PMID- 2544464 TI - Paget's disease of the nipple occurring after wide local excision and radiotherapy for carcinoma of the breast. AB - Between 1979 and 1985, 356 cases of Stage I or II breast carcinomas have been treated at Westminster Hospital with breast conservation. Of these patients, 338 underwent local excision and radiotherapy and there have been 38 local recurrences. Three of these have been as Paget's disease of the nipple occurring 16, 8 and 5 months following radiotherapy. In each case the nipple appearances were attributed to postradiotherapy changes; the true diagnosis was not made for an average of 19 months. The increasing use of conservative breast management for early breast tumours permits the appearance of Paget's disease as a significant proportion of local recurrences. Previously this has been considered to be a rare event. We recommend that Paget's disease of the nipple should be suspected early following radiotherapy in the presence of any nipple changes. PMID- 2544465 TI - Malignant mixed mesodermal tumour of the peritoneum with a complete response to cyclophosphamide. AB - We treated a 66-year-old Japanese woman with malignant mixed mesodermal tumour of the heterologous type arising from the paracolic peritoneum. The tumour contained heterologous elements in the form of cartilage in addition to carcinosarcomatous areas. This is the third case of malignant mixed mesodermal tumours originating from the peritoneum to be documented in the literature. A complete resection of the tumour was carried out but multiple lymph nodes metastases in the left neck occurred 5 months later. Cyclophosphamide in doses of 150 mg daily by mouth was initiated, with the result of a complete regression of the malignant lesions. She died of myocardial infarction 21 months after surgery. PMID- 2544467 TI - Anchorage-independent growth of RSV-transformed rat (GCA, W 12 and XC) cells. AB - Three RSV-transformed rat cell lines: GCA, W12 and XC were characterized as to their ability to anchorage-independent growth in comparison to normal rat kidney (NRK-49F) cells. Differences in the threshold density (TD) and colony forming efficiency (CFE) of the investigated cells are described. The ability of virally transformed cells to stimulation of soft agar colony formation of NRK cells in coculture assay was presented. The production of TGFs-like factors by GCA, W12 and XC cells was suggested. PMID- 2544466 TI - [Changes in the activity of alkaline phosphatase and Na K ATPase in the membrane fractions of small-intestinal epithelium in the normal state and in diarrhea]. AB - Activity of alkaline phosphatase (apical membrane) and Na+, K+-ATPase (basal membrane) is determined to decrease in the membrane fractions of intestinal epithelium in the diarrheic new born cattle. It is concluded that Na+, K+-ATPase of basal membranes play a fundamental role in Na+ loss by body with diarrhea. PMID- 2544468 TI - Direct in vitro effect of 1,25-dihydroxyvitamin D3 on islets insulin secretion in vitamin deficient rats: influence of vitamin D3 pretreatment. AB - Vitamin D3 deficiency decreased glucose-induced insulin release from isolated rat islets. In vivo, vitamin D3 treatment restored the B-cell response within 3 days; this delay suggests an effect of vitamin D3 metabolites. The effect of 1,25 dihydroxyvitamin D3 was studied in vitro on isolated islets from vitamin D3 deficient rats. When it was added to the incubation medium, it increased in a dose-dependent manner islet insulin secretion. However this effect only occurred when the vitamin D3-deficient rats received at least a single injection of vitamin D3 24 hours earlier; these results sustain the hypothesis of a direct but delayed in vitro stimulation of B cell function by 1,25-dihydroxyvitamin D3. PMID- 2544469 TI - Lack of diabetogenic action of calcitonin in subjects affected by insulinoma. AB - Calcitonin has been shown to affect calcium handling within cells thus impairing insulin secretion and glucose tolerance in healthy subjects. In the present study we investigate the effects of calcitonin on basal and nutrients-induced plasma glucose and insulin levels variations in healthy subjects (n = 10) and in patients affected by islet cell tumor (n = 6). In healthy subjects calcitonin markedly decreased basal and nutrients-induced plasma insulin levels while in patients with islet cell tumor this calcitonin-mediated effect was lost. So we conclude that the lack of calcitonin effect upon insulin secretion in patients with insulinoma is probably due to the autonomous insulin secretion characterizing islet cell tumor. PMID- 2544470 TI - Membrane lipid alterations and Na+-pumping activity in erythrocytes from IDDM and NIDDM subjects. AB - The Na+-pumping activity of the erythrocyte plasma membrane in diabetic subjects was studied together with the lipid composition. Insulin-dependent diabetes mellitus (IDDM) patients (n = 25) were divided into young (28.1 +/- 7.4 yr old, mean +/- SD; n = 16) and old (7.17 +/- 9.8 yr old; n = 10) subjects; the age of non-insulin-dependent (NIDDM) patients was 70.7 +/- 11.5 yr (n = 10). The Na+ pumping activity, estimated from both Na+-K+-ATPase and ouabain binding, was significantly decreased in IDDM and NIDDM subjects, but its insulin sensitivity was retained only in young IDDM subjects. The total cholesterol and phospholipid content of the erythrocyte plasma membrane was lowered in IDDM subjects, and cholesterol-to-phospholipid molar ratio was significantly decreased. In NIDDM subjects the significant decreased of the two lipid components did not alter their ratio. The analysis of major phospholipid components of erythrocyte membranes revealed that only phosphatidylcholine is significantly increased in young diabetic subjects. The fatty acid composition of major phospholipid classes was significantly altered in all cases: the unsaturation index appeared to be increased in phosphatidylserine and sphingomyelin for both IDDM and NIDDM subjects and was also increased in phosphatidylcholine in the latter group. PMID- 2544471 TI - Effects of acute metabolic acidosis and alkalosis on leucine metabolism in conscious dogs. AB - To determine the effects of acute metabolic acidosis and alkalosis on leucine metabolism in vivo, mongrel dogs were infused with [1-14C]leucine for 8 h, along with NaCl, HCI, or NaHCO3 over the last 4 h. Arterial pH did not change from the basal value during NaCl infusion but decreased (P less than .01) and increased (P less than .01) during HCl and NaHCO3 infusions, respectively. Total leucine carbon entry did not change from the basal value during saline infusion but increased (P less than .01) with acidosis and decreased (P less than .05) with alkalosis. Compared with saline controls, acidosis increased (P less than .01) leucine oxidation. During alkalosis decreased (P less than .01) leucine oxidation. During acidosis, total plasma essential and nonessential amino acid concentrations increased (P less than .05), whereas during alkalosis, total plasma essential and nonessential amino acid concentrations decreased (P less than .05). These studies suggest that acute alterations in arterial pH may affect the regulation of protein metabolism in vivo and must be considered in the interpretation of results from experiments in which alterations of acid-base homeostasis may have occurred. PMID- 2544472 TI - Interferon-gamma induces transcription and differential expression of MHC genes in rat insulinoma cell line RINm5F. AB - We have reported that enhanced levels of class I major histocompatibility complex (MHC) antigen are expressed throughout the islets of prediabetic and newly diabetic BB rats and that the endocrine cells of the islet remained class II negative. In this study we investigated the molecular biology of lymphokine induced expression of the class I and II MHC genes in subclones of the rat insulinoma cell line RINm5F. Treatment of a particular subclone of RINm5F cells (which are normally class II negative, class I low expressors) with crude lymphokine preparation or various doses of recombinant interferon-gamma resulted in enhancement of MHC class I antigen expression but no detectable induction of class II antigen expression. This enhancement of class I antigen expression was a dose-dependent phenomenon and was preceded by a dose-dependent increase in class I-specific RNA. Both class I and II genes were induced at the transcriptional level, as determined by Northern blotting and in vitro nuclear transcription assays, but exhibited strikingly different induction kinetics. Supernatants from concanavalin A-stimulated splenocytes had a similar class I-restricted inductive effect on MHC gene expression. This subclone of RINm5F cells, which exhibits a class I lymphokine response-positive, class II response-negative phenotype, 1) mimics the behavior of beta-cells in the prediabetic and newly diabetic pancreas and 2) represents a valuable system for probing the similarities and differences in the lymphokine-mediated induction pathways for class I and II MHC genes. PMID- 2544473 TI - Development of a method for isolation of islets from human fetal pancreas. AB - Three methods for the preparation of islets from human fetal pancreas (17.4 +/- 1.2 wk gestational age) were compared. In each method, the pancreases were minced and followed by 1) no collagenase digestion, 2) 5 min of collagenase digestion, or 3) 14 min of collagenase digestion. The culture conditions prevented adherence of the fragments. Culture for 6-7 wk of minced fetal pancreas without collagenase digestion resulted in fragments that were a mixture of cells positive for insulin or glucagon, ducts, necrotic debris, and other unidentified cells with complete degeneration of the acinar cells. Culture of minced pancreas digested for 5 min with collagenase resulted in fragments that superficially appeared to be islets but did not have the size characteristics of human fetal islets and contained fibrous and duct elements not seen in islets. Culture of minced pancreas digested for 14 min with collagenase resulted in islets that were released into the medium and harvested by picking. These islets were morphologically similar to islets of the intact human fetal pancreas and isolated islets from rat neonatal pancreas. These islets and fragments were viable for at least 7-8 wk in culture. PMID- 2544474 TI - [Response to cholera toxin of 2 epithelial intestinal cell lines. Effect of Saccharomyces boulardii]. AB - Cholera toxin acts in vivo by activating intestinal adenylate cyclase. This study was designed to determine (1) whether normal rat epithelial intestinal cell lines (IRD 98 and IEC 17) respond to cholera toxin (CT) by an increased concentration of cyclic AMP and (2) whether the yeast Saccharomyces boulardii, which reduced CT induced secretion of water and electrolytes using the isolated jejunal loop technique, has an effect on these models. The cAMP concentration evaluated in cells exposed to Saccharomyces boulardii and to cholera toxin (1 microgram/ml for 90 min) was compared to the concentration of cAMP obtained in control cells without yeast. Prior exposure of IRD 98 and IEC 17 cells to Saccharomyces boulardii, reduced CT-induced cAMP by 50 p. 100. This effect disappeared after destruction of the yeast by heating. Results show that the IRD 98 and IEC 17 cells are good models for in vitro investigation of the effects of cholera toxin. Our results suggests that Saccharomyces boulardii prevents the water and electrolyte secretion induced by cholera toxin. PMID- 2544475 TI - [Automated microscopic image analysis and the prognosis of preneoplasms and carcinomas of the mammary gland]. AB - Studies of preneoplasias and carcinomas of the mammary gland have been conducted by means of automated microscopic image analysis, on the basis of previous results as well as with references to the international literature, for 2 purposes: 1. Determination in the context of a clinical follow-up study of the individual carcinoma risk for patients with proliferative fibrocystic breast disease (mastopathy) and 2. Preparation of an objective automated grading of ductal breast carcinomas for better assessment of prognosis. Against the background of the assumption that the majority of carcinomas and precancerous lesions of the breast originate from the terminal ductal lobular unit, an effort is made to determine, independent severity of the mastopathy, the biological valence of solid, cribriform, and papillary ductal epithelial proliferations (no, possible or inevitable preneoplasia). Proliferation patterns without and with atypical features are checked for their similarity with intraductal and invasive carcinomas (similarity principle) and are additionally examined for differences, depending on localization and distance from tumour (topological principle). Automated histological tumour grading is to distinguish with greater subtlety within the large heterogeneous group of moderately differentiated carcinomas and is to objectify and thus facilitate the difficult task of delimitation of moderately and poorly differentiated carcinomas. Nuclear grading will be the major basis for classification along these lines. Objectified and reproducible tumour grading is believed to be extremely helpful in better prognostication of breast cancer. PMID- 2544476 TI - [Epithelial growth factor receptors (EGF-R) and DNA flow cytometry in addition to lymph node and hormone receptor status as prognostic factors in primary breast cancer]. AB - The presence of receptors for epidermal growth factor (EGF-R), the percentage of cells in S-phase and DNA-ploidy measured by flow-cytometry are new prognostic factors in breast cancer. In order to investigate the relationship between EGF-R, S-Phase/DNA-ploidy and clinical data, we measured these parameters in 68 tumors from patients with operable breast cancer. 11 Patients had positive EGF-R's (specific binding greater than 10 fmol/mg membrane-protein) and 37 patients had a positive S-Phase (more than 5% of cells in S-Phase). 10 of 44 carcinomas of ductal type were EGF-R positive, while only 1 of 18 lobular-type tumor displayed positive EGF-R's. We found significant inverse correlations between EGF-R's and estrogen receptors (p less than 0.00001) and between EGF-R's and age (p less than 0.0001): EGF-R positive tumors were found more often in young patients and in patients with estrogen-receptor-negative tumors. The EGF-R's and flow-cytometric data did not correlate with tumor size and/or nodal status. There was no correlation between EGF-R and S-Phase/DNA-ploidy in the breast cancer specimens suggesting that these two prognostic parameters are independent from each other. PMID- 2544477 TI - [Hormone dependence of malignant ovarian tumors--an in vitro model]. AB - The steroid hormone receptor content of 32 malignant ovarian tumors was compared with the in vitro effectiveness of 4 hydroxytamoxifen (OH-TAM) and medroxy progesterone acetate (MPA) tested in the Human Tumor Colony Forming Assay (HTCFA). The sensitivity for the receptor determination was 5 fmol/mg cytosol protein. Estrogen receptors (ER) and progesterone receptors (PR) were found in 15 (47%) and 13 (41%) of the tumors respectively. As standard criteria for the HTCFA, a minimum of 30 colonies with a diameter of more than 60 microns and 100 microns was used in the control group. The in-vitro sensitivity of ovarian tumors to OH-TAM and MPA was independent on the ER or PR content, and amounted to 9% for OH-TAM and 6% for MPA. However, all 12 ER-PR-tumors proved resistant to OH-TAM and MPA. 18 ovarian tumors showed a sufficient colony growth, even in the size class exceeding 100 microns. With a minimum colony size of 60 microns and 100 microns, 17% and 33% respectively were sensitive to OH-TAM. A similar effect on the proliferative capacity of the Tumor Colony Forming Units (TCFUs), unrelated to PR, was observed with MPA. Dependent on colony size, we found an increasing sensitivity against MPA from 11% to 22%. The in-vitro effectiveness of both OH TAM and MPA in the clonogenic assay of malignant ovarian tumors was certainly not as potent as suggested by the results obtained in biochemical steroid hormone receptor analysis. To prove the hormonal response in the HTCFA, it is necessary to determine number and size of the colonies as an expression of their proliferative potential. PMID- 2544478 TI - Model of calcium channel inactivation: a qualitative analysis. AB - A simple model of calcium channel inactivation has been developed, based on the accumulation of calcium ions at the inner mouth of the channel and on their binding to a receptor which inactivates the channel. A qualitative analysis has shown that upon an appropriate choice of parameters corresponding to the cell structure and to kinetic properties of its components, the calcium dependent inactivation and that assumed to be voltage dependent can both be emulated. The model suggests that the supposed variety of calcium channels might be explained by quantitative differences in nonlinear interactions of the channels with other cell components. PMID- 2544479 TI - The steady state stability criterion of a simple model of calcium channels. AB - A simple known model of calcium inactivation is described and qualitatively analysed. Stability conditions at the level of a stationary state with respect to some small perturbations in the concentration of Ca2+ ions are analysed from the point of view of the Prigogine non-equilibrium thermodynamics. Possible internal fluctuations in Ca2+ ion concentration are discussed as connected with fluctuations of the potential energy of interaction between calcium ions and the binding sites. PMID- 2544480 TI - Ontogenic differentiation of pig atrial and ventricular myosin. AB - Myosin was isolated from pig atrial and ventricular myocardium during postnatal development and Ca2+-ATPase was determined and myosin light chains were analysed by electrophoresis in sodium dodecylsulfate polyacrylamide gel. During ontogenesis ATPase activity of ventricular myosin remains virtually unchanged, whereas that of atrial myosin increases. The patterns of myosin light chains of atrial and ventricular myosin differ from each other, but the individual pattern remains unchanged during the development. PMID- 2544481 TI - Lower autolytic activity in a homogeneous methicillin-resistant Staphylococcus aureus strain compared to derived heterogeneous-resistant and susceptible strains. AB - It has been proposed that in addition to production of a penicillin-binding protein with low affinity for beta-lactam antibiotics, control of autolysin activity is involved in the mechanism of staphylococcal methicillin resistance. A homogeneous methicillin-resistant Staphylococcus aureus strain (DU4916) had lower rates of unstimulated, NaCl- and Triton X-100-stimulated autolysis, and daptomycin (LY146032)-induced lysis than a heterogeneous methicillin-resistant strain (DU4916-K7) and a methicillin-susceptible strain (DU4916S) derived from DU4916. PMID- 2544482 TI - Monoclonal antibodies against alpha toxin of Clostridium perfringens. AB - Ten distinct monoclonal antibodies (MAbs) against alpha toxin of Clostridium perfringens were produced by the fusion of SP2/O with spleen cells of mice immunized with alpha toxoid, and alpha toxin mixed with or without ethylenediamine-tetraacetate (EDTA). The antibody activity was evaluated by antigen-binding activity in an enzyme linked immunosorbent assay (ELISA), by phospholipase C (PLC)-neutralizing activity using both egg yolk lecithin and p nitrophenylphosphoryl-choline (PNPPC) hydrolysis reactions and by anti-lethal activity in mice. Since the toxin-neutralizing activities of each MAb were not parallel, it has been suggested that the three biological activities may not be located in the same site in the toxin molecule. This report also describes the development of a simple purification of the toxin by affinity chromatography and a sensitive immunoassay for quantitation of the toxin using the monoclonal antibody. PMID- 2544483 TI - Stimulation of growth and sporulation of Clostridium perfringens by its homologous enterotoxin. AB - C. perfringens enterotoxin shortened the lag phase and time of onset of sporulation of the same organism in a dose-dependent manner. The toxin stimulated macromolecular synthesis of pre-exponential phase cells. PMID- 2544484 TI - A cation/proton antiport activity in Acholeplasma laidlawii. AB - Sealed membrane vesicles of Acholeplasma laidlawii were obtained by controlled lysis of carotenoid-rich intact cells. An imposed delta pH was created by loading membrane vesicles or intact Acholeplasma laidlawii cells with 0.25 M NH4Cl and diluting them into 0.25 M choline chloride. The passive efflux of NH3 from the membrane vesicles or cells resulted in the creation of a delta pH (inside acid) that could be visualized by the quenching of the fluorescence of the weak base acridine orange. Whereas with isolated membrane vesicles, the fluorescence was dequenched by the addition of Na+, with intact cells, K+ in addition to Na+ was required. These results strongly suggest a Na+/H+ exchange activity that in intact Acholeplasma laidlawii cells is K+-dependent. The possible role of the Na+/H+ exchange activity in pH homeostasis at the more alkaline pH range, as well as in the extrusion of excess Na+ from the cells is discussed. PMID- 2544485 TI - [Cloning the genetic determinant of alpha-hemolysin and obtaining a series of insertional mutations in this determinant by the transposon Tn1000]. AB - Functionally active genetic determinant of alpha-hemolysin was cloned. Hemolytic plasmid pHly195 was used as a donor of the determinant and pBR322 plasmid served as recipient. Cloning was done with a help of HindIII restriction endonuclease. The recombinant plasmid obtained represents pBR322 plasmid with the built-in fragment of 7.4 kb containing genes of functionally active determinant of alpha hemolysin. Restriction map was constructed using HindIII, EcoRI, BamHI and SalI restriction endonucleases. Insertional mutagenesis was carried out with the help of the Tn1000 transposon. Plasmid DNAs were isolated from insertional mutants of Hly- phenotype and treated with EcoRI, SalI and BamHI. On the basis of the sizes of restriction fragments of the mutant plasmid DNAs localization and orientation of insertions of Tn1000 into the cloned determinant of alpha-hemolysin were determined. PMID- 2544486 TI - Geriatric falls: assessing the cause, preventing recurrence. AB - Falling represents a common and dangerous problem for the elderly. As a result, fall prevention has become a focus of clinical attention. This article examines why older people fall, what factors place persons at risk for falling, and how to prevent falls. PMID- 2544488 TI - Small changes in heart rate following alpha-1 adrenoceptor stimulation in the anaesthetized dog. AB - In a previous work (1) we observed a weak alpha-1 adrenoceptor mediated chronotropic effect in anaesthetized dogs: the intracoronary injection of 100 micrograms of amidephrine, an alpha-1 agonist, increased heart rate by 2.5 +/- 0.8 bpm (mean +/- SEM). Since these experiments had been performed in the presence of alpha-2 blockade with yohimbine, one could argue that alpha-1 adrenoceptors had been partially blocked as well. To test for this possibility 5 additional experiments were performed with the same protocol, just omitting yohimbine administration. The chronotropic effect of amidephrine was larger (6.2 +/- 1.9 bpm after i.c. injection of 100 micrograms), but the difference was not significant. This confirms our earlier finding that alpha-1 adrenoceptors are not involved in heart rate control of the anaesthetized dog. PMID- 2544487 TI - Sporadic aniridia and Wilms' tumor: visual function evaluation of three cases. AB - The visual function of three infants with sporadic aniridia associated with Wilms' tumor and a deletion of the short arm of chromosome 11 was evaluated with electrophysiological tests. The patients presented nystagmus and photophobia. The electroretinograms (ERGs) were normal, as in other sporadic cases, but at variance with the familial cases. The latency of the flash visual evoked potentials (FVEPs) became shorter with time but remained longer than in age matched controls, suggesting a delay in maturation of the nervous system. Poor visual function in our cases did not appear to be the result of gross retinal anomalies, as shown by the normal ERG, nor of an hypoplasia of the optic nerve. Contact lenses that provide an artificial pupil decreased photophobia and nystagmus and are therefore highly recommended to increase patient comfort. PMID- 2544489 TI - Carcinosarcomas and mixed Mullerian tumors of the fallopian tube. AB - Four cases of carcinosarcoma and mixed mullerian tumors of the fallopian tube are presented. Each patient presented with abnormal bleeding and a pelvic mass. All underwent total abdominal hysterectomy, bilateral salpingo-oophorectomy, staging, and cytoreduction. Disease was limited to the pelvis in two patients, analogous to FIGO stage IIB ovarian carcinoma; the other two patients had upper abdominal disease, analogous to FIGO stage III. The primary tumors were intraluminal and papillary. There were equal amounts of carcinoma and sarcoma in three tumors; in one, sarcoma constituted only a small intraluminal focus. The sarcoma was predominantly homologous, with foci of heterologous elements present in three tumors. Adjuvant therapy consisted of pelvic radiation in two patients. One patient died of inanition within one year of diagnosis. The other patient, who had the small focus of sarcoma within a stage IIB carcinoma, had an 11-year disease-free interval before retroperitoneal recurrence of carcinoma. Two patients received chemotherapy. A stage IIB patient, after pelvic radiation, received Cytoxan and Adriamycin; she is clinically free of disease after 6 years. A stage III patient lived over 3 years after treatment with multiple agents; she responded to Cytoxan and cis-platinum before suffering a systemic relapse and death. PMID- 2544490 TI - High-risk metastatic gestational trophoblastic disease: further stratification into two clinical entities. AB - Forty-two of sixty-seven patients (62.7%) treated for high-risk metastatic trophoblastic disease achieved and maintained complete remissions. The survival rate was significantly improved in those patients with scores lower than 8 according to a modification of the World Health Organization (WHO) prognostic scoring system. A low score was associated with a higher probability of response to single-agent therapy, although the difference was not statistically significant. The score, however, was significantly associated with response to multiagent chemotherapy with methotrexate, actinomycin D, and cyclophosphamide (P = 0.0004). Therefore, future trials of new combinations of chemotherapy in high risk patients should be stratified according to the patients' prognostic scores. PMID- 2544491 TI - Successful salvage therapy of resistant gestational trophoblastic disease with etoposide, methotrexate, dactinomycin, vincristine, and cyclophosphamide. AB - A case of poor prognosis metastatic trophoblastic disease resistant to chemotherapy with methotrexate, dactinomycin, and cyclophosphamide is presented. The patient responded promptly to etoposide-based combination chemotherapy and has remained in complete remission since completion of therapy. PMID- 2544492 TI - [Mammography and ultrasound in the diagnosis of clinically occult breast cancer]. AB - 105 biopsies of nonpalpable breast lesions were performed because of suspect X ray findings. 68 lesions were preoperatively marked under X-ray control, 37 under ultrasound control. Histological examination revealed invasive and in situ carcinomas in 8 and 13% of the cases, respectively. The sensitivity of X-ray mammograms was superior to ultrasound, whereas the predictive value of ultrasound in the presence of suspect findings was superior to x-ray examination. However, because of its low sensitivity, sonographic examination cannot be recommended as a screening procedure in the diagnosis of nonpalpable breast lesions. PMID- 2544493 TI - [Pain modulation in labor by endogenous opiates]. PMID- 2544494 TI - [Experiences with DHPG (ganciclovir)--treatment of cytomegalovirus retinitis in AIDS]. AB - Nine patients with unilateral cytomegalovirus (CMV) retinitis were treated with intravenous infusions of the new virustatic drug DHPG (Ganciclovir). The induction dose was 10 mg/kg body weight per day (2-5 weeks). In six cases, a maintenance dose was given thereafter of 5 mg/kg body weight per day. After the induction dose, five patients had visual improvement and satisfactory cicatrization. In one patient, PVR (proliferative vitreous reaction) developed with total tractional retinal detachment. More ocular complications were seen while on longterm therapy: relapses during discontinuation because of leukopenia (three times in two patients), breakthrough (= relapse during maintenance therapy) (one case), serous retinal detachment (one case), and optic atrophy (two cases). The complications caused blindness in two further patients. Only one patient has tolerated maintenance therapy for 22 weeks without having any complications. One patient wanted to have therapy suspended and has remained free of relapse for 28 weeks while on cytostatic therapy. The eyes of two deceased patients were examined histopathologically, immunhistochemically, and ultrastructurally and the findings compared with those of an untreated case. Given at an early stage and without discontinuation, DHPG is an effective means of preventing or delaying blindness caused by cytomegalovirus retinitis in AIDS patients. The directives for an optimal dosage are subject to further prospective randomized clinical studies. PMID- 2544495 TI - [The peroxidase content of human tears]. AB - The peroxidase-(POD)-thiocyanate-hydrogenperoxide-system is a well-known antibacterial system, which has been demonstrated to exist, for example, in milk and saliva. Earlier investigations by van Haeringen et al. established a POD level in human tears of 10(3) units/l, yet the thiocyanate concentration was only about 0.2 mmol/l. Therefore van Haeringen et al. excluded the existence of a POD thiocyanate-hydrogenperoxide antibacterial system in human tears because of the insufficient amount of thiocyanate in the tears examined. Instead of thiocyanate halides such as J- can also complete the POD hydrogen peroxide system as electron donors. Sufficient amounts of iodide can be expected after the application of iodine-containing eye drops or after local treatment with iodine-containing brine, as done in Bad Hall in Austria. Therefore, the above mentioned antibacterial system may be of importance if the POD-level is high enough (greater than 250 units/l). We investigated 22 tear samples from healthy persons: the POD levels were below 20 units/l in 19 cases; in 3 cases the POD concentration was found to be between 20 and 50 units/l. Therefore, in normal human tear fluid, not only the amount of thiocyanate but also the concentration of POD is too low for effective antimicrobial activity of the peroxidase thiocyanate-hydrogenperoxide system. It is so far not known whether this system is effective under pathological conditions. PMID- 2544496 TI - [Gangliosides in the treatment of polyneuropathies. Satellite symposium of the 61st annual meeting of the German Society for Neurology. Frankfurt, 21 September 1988. Abstracts]. PMID- 2544497 TI - Quick screening of plasmid deletion clones carrying inserts of desired sizes for DNA sequencing. AB - Plasmids pWQX001 and pWQX005, constructed from pGEM-4 with an insert of 6.5 kb, were unidirectionally digested with exonuclease III and exonuclease VII. The DNA digests were ligated and used to transform competent cells of Escherichia coli DH5-alpha. The size of the deletion plasmid carried by each transformant was estimated through agarose gel electrophoresis of crude lysates without any purification of the plasmid DNA. Colonies carrying plasmid DNAs with different deletions of the insert were grown and their DNAs were purified through a miniprocedure. The size of each purified plasmid DNA was determined accurately after linearization of the plasmid with an appropriate restriction endonuclease. The remainder of the DNA preparation was sufficiently pure to be sequenced using Sanger's dideoxynucleotide chain termination method. An easy, quick procedure is described for the preliminary selection of templates for DNA sequencing after construction of deletion clones of recombinant plasmid DNA using exonuclease III and exonuclease VII. This procedure permits a rapid screening of large numbers of colonies and selection of those carrying plasmid DNAs with inserts of the desired sizes for sequencing. This procedure does not require purification of the deletion plasmid DNA. PMID- 2544498 TI - Active and latent collagenase activity during reversal of hepatic fibrosis in murine schistosomiasis. AB - In schistosomiasis mansoni, eggs trapped in portal venules of the liver induce a granulomatous inflammatory reaction and subsequently fibrosis. Upon specific schistosomicidal treatment, however, fibrosis undergoes a marked reversal. The features of this process as regards collagen metabolism were examined in mice using praziquantel as the schistosomicidal drug, which was given 68 days after infection. It was found that collagenase activity in the liver, for both latent and active forms, increases rapidly until 5 days after treatment as compared to nontreated infected mice and then diminishes gradually. The hydroxyproline content in liver and (24-hr) urine are also modified, their values decreasing progressively up to 72 days after treatment without, however, reaching values observed in normal mice. Worm death appears to be the triggering stimulus of these events. PMID- 2544499 TI - A 5-year experience of lipiodolization: selective regional chemotherapy for 200 patients with hepatocellular carcinoma. AB - Between 1982 and 1987, selective regional cancer chemotherapy using Lipiodol plus an anticancer drug (lipiodolization) was prescribed for 200 patients with hepatocellular carcinoma. One hundred forty-nine patients were given lipiodolization alone, and the remaining 51 underwent hepatic resection following lipiodolization. The grades of deposits of the oily contrast medium in the neoplastic tissue seen on the plain X-ray correlated well with the antitumor effect. In the resected specimens of 17 patients treated with lipiodolization prior to surgery, concentrations of adriamycin in the malignant liver tissues were 13.2 +/- 18.2 micrograms per gm, whereas the adjacent liver parenchyma contained 1.4 +/- 2.0 micrograms per gm of adriamycin; the difference was statistically significant (p less than 0.05). In the 149 patients treated with lipiodolization, 1-, 2-, 3- and 4-year survival rates were 56.1, 28.9, 17.3 and 7.4%, respectively. Thus, lipiodolization was considerably more effective, compared to the results achieved by hepatic artery ligation and cannulation into the hepatic artery for patients in Stages I and II. In this sequential nonrandomized study, the survival rates for patients undergoing hepatic resection were superior to those for patients in Stage I and treated with lipiodolization. The significant difference appeared to depend on incomplete killing of tumor cells, which are most often present in the fibrous capsule, by lipiodolization. We conclude from these data that lipiodolization is an effective treatment for hepatocellular carcinoma when the tumor is not curatively resectable. When the clinical status is good, then surgery is warranted. PMID- 2544500 TI - Phenotypic characterization of non-haemopoietic small cell tumours of childhood with monoclonal antibodies to leucocytes, epithelial cells and cytoskeletal proteins. AB - Recently, great interest has been shown in the histological identification of small cell tumours of childhood--nephroblastoma (Wilms' tumour), neuroblastoma, rhabdomyosarcoma and Ewing's sarcoma--using immunohistochemical methods. However, several antigens operationally specific for leucocyte typing in blood and marrow are also expressed on cells of epithelial and neural origin. We undertook phenotypic characterization of 17 non-haemopoietic small cell tumours of childhood using a panel of 30 monoclonal antibodies to leucocyte, epithelial and cytoskeletal antigens using a sensitive alkaline phosphatase-anti-alkaline phosphatase technique on cryostat sections of fresh tumour. Our results demonstrated frequent expression of the leucocyte-associated antigens CD10 (CALLA), CD9 (p24) and CDw32 (FcRII) in these small cell tumours and occasional expression of MHC class II (HLA-DR) and HNK-1 antigens. However, the leucocyte associated antigens CD45 (leucocyte common), CD22 (pan B-cell), CD11b (C3bi receptor), CD15 (Lewisx) or CDw42 (platelet gp Ib) were not detected on any tumour. Aberrant expression of desmin, neurofilament and UJ13A antigen was found in nephroblastoma and of epithelial-associated markers (CIBr17 and 43-9F) in neuroblastoma. Our results also demonstrated broad reactivity in frozen section with two monoclonal antibodies specific for melanoma (NKI/C-3) or epithelial cells (OM-1) in paraffin sections. Hence, it is necessary to include monoclonal antibodies to CD45 and pan-epithelial antigens, e.g. LP34 (cytokeratin) or HEA125 for the precise immunohistochemical identification of small round cell malignancies of childhood. PMID- 2544501 TI - The histogenesis of mammary and extramammary Paget's disease. AB - The histogenesis of mammary and extramammary Paget's disease has been studied by immunohistochemical staining of paraffin-embedded tissue using a panel of epithelial cell markers, which react with secretory or ductal epithelium, but not stratified epithelium. These markers included a monoclonal antibody E29 to epithelial membrane antigen EMA, the cytokeratin marker CAM 5.2 and three new monoclonal antibodies raised to human milk fat globule membrane (LICR-LON-TW19 and H.10.A) and a human bladder cell cancer line (3.77). The findings demonstrate that both mammary and extramammary Paget's disease are of epithelial cell origin and share antigens expressed by simple epithelia. Some antigens, such as EMA and low molecular weight cytokeratins are consistently present in both diseases, whereas other antigens, identified by H.10.A and TW19 are found more frequently in cases of extramammary Paget's disease. This panel of monoclonal antibodies also proved useful in distinguishing Paget's disease from pagetoid melanoma and clear cell Bowen's disease. PMID- 2544502 TI - Malignant granular cell tumour with intraperitoneal dissemination. PMID- 2544503 TI - Mammary carcinoma arising in benign axillary epithelial lymph node inclusions. PMID- 2544504 TI - Glomus tumours: an immunohistochemical profile of 11 cases. AB - We studied 11 glomus tumours immunohistochemically, with a panel of connective tissue and epithelial markers. Most tumours contained small nerve fibres located in connective tissue septae between groups of glomus cells, thus accounting for the frequent occurrence of pain associated with glomus tumours. All tumours stained positively for muscle-specific actin and vimentin. Immunostaining for high and low molecular weight cytokeratins, desmin, myoglobin, S-100 protein, neurofilaments and Factor VIII related antigen was negative. Our findings confirm and amplify the proposed smooth muscle histogenesis of glomus tumours. This immunohistochemical profile may be of diagnostic value in the differential diagnosis of atypical glomus tumours. PMID- 2544505 TI - Primary intra-osseous glomus tumour. PMID- 2544506 TI - Metaplastic carcinomas of the breast. I. Matrix-producing carcinoma. AB - The clinical and pathologic features of 26 examples of a histopathologically distinct form of metaplastic carcinoma of the breast are reported. All neoplasms had overt carcinoma with direct transition to a cartilaginous and/or osseous stromal matrix without an intervening spindle cell zone or osteoclastic giant cells. Therefore, we designate this distinctive form of metaplastic carcinoma as "matrix-producing carcinoma" (MPC). All patients were women, the average age was 58 years, and all patients were eligible for a minimum of 5 years follow-up (mean follow-up period, 8.6 years). Twenty-three patients were treated by a form of mastectomy and three were treated by local excision. The 5-year survival rate for patients following mastectomy or partial mastectomy was 70%, contrasted with 50% for patients treated by local excision. The cumulative 5-year survival rate for MPC was 68%. All of the nine lesions that recurred did so within 2.5 years of initial therapy. Eight of these patients (89%) died from tumor within 4 years of initial therapy. The ninth was alive at last contract. Radiation and chemotherapy were of limited effectiveness. Significant features of the neoplasm associated with progression were large size, diffuse cellularity of the stromal matrix, and atypical cartilaginous metaplasia. Ultrastructural examination of one case and immunohistochemical evaluation of 12 cases revealed MPC to have myoepithelial characteristics. PMID- 2544507 TI - Classical and cellular (atypical) congenital mesoblastic nephroma: a clinicopathologic, ultrastructural, immunohistochemical, and flow cytometric study. AB - Sixteen cases of congenital mesoblastic nephroma (CMN) were studied. The tumors showed variable patterns of growth, degrees of cellularity, and mitotic activity. Six tumors had the classical pattern of CMN, seven were of the cellular or atypical variant and three showed combined features. The mean ages at presentation were 16 days, 5.3 months, and 2.3 months, respectively. Average size and weight were 5.1 cm and 94 g for classical CMN, 9.1 cm and 620 g for cellular CMN and 10.5 cm and 150 g for combined tumors. Cyst formation, hemorrhage and necrosis were confined to cellular CMNs and to cellular areas of combined CMNs. Mitotic activity ranged from 0 to 1/10 high-power fields (HPFs) in classical tumors to 25 to 30/10 HPFs in cellular tumors. Clear cell sarcoma-like areas were observed in three neoplasms. In ten cases there was invasion of perirenal fat; in one case each, invasion of the psoas muscle, renal vein wall, and renal vein lumen was observed. Ultrastructural and immunohistochemical studies showed features consistent with myofibroblastic differentiation. Flow cytometric analysis revealed euploidy in one classic CMN, one cellular CMN and in classic areas of a combined CMN; cellular areas of the latter tumor were aneuploid. All patients with follow-up were alive without evidence of disease after a mean period of 5 years following nephrectomy alone. No correlation was observed between the pathologic features assessed and the biologic behavior of these neoplasms. PMID- 2544508 TI - Extrarenal Wilms' tumor: an analysis of four cases. AB - During a review of Wilms' tumor, four located external to the kidney were identified. Patient age ranged from 7 months to 4 years; three were female. One neoplasm was located in the parametrial connective tissue to the left of the uterus; both kidneys were radiographically normal. Three neoplasms were located in the right retroperitoneum adherent to the surface of the ipsilateral kidney, but separated from the parenchyma by a thick fibrous capsule. Two were attached to the upper pole, while the third was attached to the midportion of the kidney. Radiologic studies showed displacement of all three kidneys, but intravenous pyelogram (IVP) revealed no calyceal distortion. All four neoplasms were favorable histology Wilms' tumor: one was monophasic epithelial type, one was monophasic blastemal type, and two had a mixture of stromal, epithelial, and blastemal tissue. No teratomatous elements were present. Immunoperoxidase staining for cytokeratin (AE-1, AE-3, CAM 5.2), vimentin, and epithelial membrane antigen (EMA) showed the strongest focal positive staining of tubular structures with CAM 5.2, and slight staining with EMA. Staining reaction to vimentin was variable, but negative in most areas. Three tumors extracted from paraffin were diploid by quantitative flow cytometric DNA analysis; in one case, flow cytometry could not be performed. Clinical follow-up from 2 years to 6 years showed all children to be alive without evidence of disease. Based on the similarity to conventional renal Wilms' tumor, these findings support the hypothesis of displaced mesonephric/metanephric rests for the origin of extrarenal Wilms' tumor. PMID- 2544510 TI - Detection of beta-thalassemia and hemoglobin E genes in Thai by a DNA amplification technique. AB - Enzymatic DNA amplification and polyacrylamide gel electrophoresis, which demonstrate different sizes of DNA fragments, were used to detect the common mutations causing beta-thalassemia and hemoglobin (Hb) E in Thai people. The 4-bp deletion at codons 41 and 42 can be detected directly by polyacrylamide gel electrophoresis and ethidium bromide staining. Whereas the nonsense mutations at codon 17 (AAG----TAG) and Hb E (GAG----AAG at codon 26) were detected after digestion of the amplified DNA with the enzymes MaeI and MnlI, respectively. PMID- 2544509 TI - Three novel partial deletions of the low-density lipoprotein (LDL) receptor gene in familial hypercholesterolemia. AB - The low-density lipoprotein (LDL) receptor genes from 18 unrelated Japanese heterozygotes and 1 homozygote with classical familial hypercholesterolemia were analyzed by Southern blot hybridization using fragments of the human LDL receptor cDNA as probes. Four different deletion mutations were detected among 20 mutant LDL receptor genes (20%); they were characterized by restriction mapping. None of these mutations has previously been reported in Caucasian patients with FH: three of the mutations were novel and one was similar to the deletion mutation of FH Tonami described previously in Japanese patients. In three of the four deletion mutations, the rearrangements were related to intron 15 of the LDL receptor gene, in which many Alu sequences exist. The data suggest that a wide range of molecular heterogeneity exists even in major rearrangements resulting in deletions in the LDL receptor gene. The data also support the hypothesis that there are preferential sites within the LDL receptor gene for major rearrangements resulting in deletions. The possibility that a higher frequency of deletion mutations occurs in classical FH than previously suspected is discussed. PMID- 2544511 TI - Loss of heterozygosity on chromosome 10 in human glioblastoma multiforme. AB - Recessive mutations, revealed by loss of the wild-type allele, have been associated with the development of a variety of cancers in children and adults. Polymorphic chromosome 10 markers were used to screen paired tumor and lymphocyte DNA samples in 13 patients with glioblastoma multiforme. Ten patients showed loss of constitutional heterozygosity in the tumor samples. This finding suggests that a recessive gene involved in the development of glioblastoma multiforme is present on chromosome 10. PMID- 2544512 TI - The role of T cells in mouse cytomegalovirus myocarditis. AB - BALB/c mice infected with murine cytomegalovirus (MCMV) developed myocarditis. Athymic nu/nu mice infected with the virus did not develop myocarditis, in contrast to heterozygous T-cell competent nu/+mice. MCMV-infected BALB/c mice given cyclosporin A(CsA) a drug which inhibits the activation of T cells, showed a delay in the development of myocarditis relative to CsA-untreated mice infected with MCMV. However, BALB/c mice infected with MCMV, regardless of CsA treatment, developed both anti-MCMV antibodies and autoantibodies. Nu/nu mice infected with MCMV did not produce the anti-MCMV antibody response or the multiple autoantibody response which was observed in nu/+ MCMV-infected mice. Both nu/nu and CsA treated animals displayed greater organ distribution of viral antigen than control MCMV-infected animals. These results suggest that the presence of a thymus is required for both the development of myocarditis and the multiple autoantibody response, which includes autoantibodies to cardiac muscle, and that CsA immunosuppression does not abrogate either myocarditis or the antibody response in mice following MCMV infection. PMID- 2544513 TI - Regulation of IFN-gamma-induced host cell MHC antigen expression by Kirsten MSV and MLV. I. Effects on class I antigen expression. AB - We have reported previously that the Kirsten murine sarcoma virus (Ki-MSV) that carries the v-Ki-ras oncogene prevents C3H10T 1/2 fibroblasts from being able to respond to interferon-gamma (IFN-gamma) with the expression of the class II major histocompatibility complex (MHC) antigen, H-2A. In this report we investigate further as to whether MSV or its parent virus Kirsten murine leukaemia virus (Ki MLV) is able to reduce host class I MHC antigen expression. The results demonstrate that class I expression is diminished in MSV-infected cells over a time-course of 7 days after exposure to IFN-gamma and over a range of IFN-gamma concentrations. The optimal concentration of IFN-gamma for maximal class I expression remained unchanged. Cells infected with Ki-MLV, which failed to abolish the induction by IFN-gamma of class II antigens, also expressed lower levels of class I antigens, similar to those for cells infected with Ki-MSV, after exposure to IFN-gamma. It is likely therefore that the inhibition of class I induction is due to genetic material shared between the viruses, principally in the long terminal repeats (LTR), and hence that the mechanism of action is distinct from that responsible for the abolition of class II induction by Ki-MSV alone. Since class I antigens are required for CD8+ T cells (mainly cytotoxic T cells) to recognize (foreign) antigen this reduction in class I expression might lead to reduced visibility of infected cells to T cells and thus might contribute to the tumorigenicity of Ki-MSV-infected cells. PMID- 2544514 TI - Regulation of IFN-gamma-induced host cell MHC antigen expression by Kirsten MSV and MLV. II. Effects on class II antigen expression. AB - We have reported previously that the Kirsten murine sarcoma virus (Ki-MSV), which carries the v-Ki-ras oncogene, prevents the induction of the class II MHC antigen H-2A and reduces the induction of class I MHC antigens by interferon-gamma (IFN gamma) on C3H10T 1/2 fibroblasts. It is here shown that the abolition by the virus of H-2A expression extends also to class II antigen H-2E and that this is maintained for at least 7 days after IFN treatment. In addition no concentration of IFN-gamma tested, including supra-optimal concentrations for class I antigen expression, induced class II antigens on MSV-infected cells. Thus MSV inhibits the induction by IFN-gamma of class II MHC antigens by a mechanism other than via a change in kinetics of response to, or in the sensitivity of the cells to, IFN. The possibility that transformation by MSV could result in the (selective) outgrowth of cells unresponsive to IFN was refuted by the observation that clones of C3H10T 1/2, when infected with Ki-MSV, expressed no or dramatically reduced levels of H-2A or H-2E. One C3H10T 1/2 clone chosen for high class II expression, when transformed with Ki-MSV, did express low levels of class II antigens at optimal concentrations of IFN-gamma, suggesting that the degree of the reduction of class II expression varies with the cells that are infected. Comparison with mechanisms whereby other viruses inhibit MHC antigen display revealed an interesting possibility: IFN response sequences (IRS) identified in the virus genomes might act in trans to (down) regulate MHC antigen expression. This could be an important mechanism determining the tumourigenicity of, and immune evasion by, Ki-MSV and other viruses. PMID- 2544515 TI - Stimulation of clones specific for dsDNA or idiotypes of anti-dsDNA as a consequence of BK virus inoculation. AB - The human polyoma virus BK (BKV) has a 5 Kbp double-stranded (ds) DNA genome packed with host cell histones into a minichromosome. Two out of 5 rabbits inoculated with infectious BKV produced detectable anti-dsDNA antibodies. In two additional rabbits antibodies against idiotypes of anti-dsDNA antibodies were elicited. These findings strongly indicate the existence of immunocompetent anti dsDNA clones which are regulated, at least in part, through a network mechanism by anti-idiotype antibodies. PMID- 2544516 TI - Mechanism of innate immunity to amoebic infection in guineapigs. AB - The direct lymphocyte cytotoxicity and antibody dependent cell mediated cytotoxicity against Entamoeba histolytica was studied in 3-4 wk old and 8-10 wk old guineapigs. Both these parameters were significantly increased in splenic cells in the older animals. This might be due to maturation of the immune system or to natural infection by related parasite, leading to immunity. PMID- 2544517 TI - Electrophoretic study of the genome of human rotavirus in rural Indian community. AB - A rural cohort of 507 children, less than 3 yr of age was kept under weekly surveillance for 12 months, for morbidity due to diarrhoea. Stool specimens for rotavirus detection were obtained in 346 of the 360 diarrheal episodes occurring in 354 children and in 211 children without diarrhoeal symptoms. Rotavirus was detected by an ELISA test in 4 per cent of children with diarrhoea and 6.6 per cent of control subjects. Viral genome RNA was extracted by phenol chloroform method from all the positive samples and analysed by polyacrylamide gel electrophoresis followed by silver staining. All the strains isolated, exhibited the same electropherotype with a 'long' RNA migration pattern. This result contrasts with the genomic variability amongst rotavirus strains usually observed in different settings. PMID- 2544518 TI - Glomerular atrial natriuretic factor receptors in spontaneously hypertensive rats. AB - There are differences in the renal handling of sodium between spontaneously hypertensive rats (SHR) and their normotensive controls. We investigated whether this difference may be associated with changes in plasma and tissue atrial natriuretic factor (ANF) levels and with alterations in glomerular ANF receptors at 4, 8, 12, and 16 weeks of age. Age-matched Wistar-Kyoto (WKY) and Wistar rats were used as normotensive controls. Systolic blood pressure was higher in SHR at 8, 12, and 16 weeks, and cardiac hypertrophy was also present in these animals at 4 weeks. Plasma ANF C- and N-terminal concentrations were greater than in both normotensive groups at 8 and 16 weeks. ANF in the right atrium was higher in SHR than in WKY rats and identical to that in the Wistar group at 4 and 8 weeks. ANF in the left atrium was lower in SHR than in both control groups at week 12. No differences were found in ventricular ANF content. The density of glomerular ANF binding sites increased with age in WKY and Wistar rats but not in SHR. At weeks 8, 12, and 16, both normotensive groups had a higher density of binding sites than SHR, but binding site affinity was greater in SHR at weeks 8 and 12. After incubation with increasing concentrations of ANF, the production of cyclic guanosine monophosphate (cGMP) by isolated glomeruli from 16-week-old rats was lower in SHR than in both normotensive groups. We conclude that the development of hypertension in SHR is associated with higher plasma ANF levels and decreased glomerular ANF receptor density and glomerular cGMP production.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544519 TI - Isolation and characterization of a sodium pump inhibitor from human plasma. AB - An endogenous sodium pump inhibitor has been purified from human plasma. The purification scheme involved large scale dialysis, extraction of lyophilized dialysate by methanol followed by preparative and semipreparative scale reverse phase high-performance chromatography. A single peak of biologically active material was obtained enriched by a factor of greater than 10 billion. This material showed high chromatographic polarity, was inactivated by charring, strong acid, or alkali, and was resistant to short-term boiling. The purified material had a molecular weight between 300 and 900 g/mol and was insensitive to type I esterase and a variety of proteolytic enzymes. The purified factor inhibited the ouabain-sensitive uptake of 86Rb by human erythrocytes, binding of [3H]ouabain, and activity of dog kidney Na,K-adenosine triphosphatase (ATPase) with high affinity (less than 0.3 nM) in a time- and dose-dependent manner. Maximally effective concentrations of the digitalislike factor showed no effect on either human red blood cell Mg- or Ca-ATPase, rabbit muscle sarcoplasmic reticulum Ca-ATPase, or guinea pig stomach H,K-ATPase. The purified material is a highly potent selective inhibitor of the ion transport, receptor, and hydrolytic functions of the sodium pump. The characteristic properties of this substance suggest it may be a mammalian endogenous digitalis and may be similar to the sodium transport inhibitor detected in the plasma of volume-sensitive forms of experimental and human hypertension. PMID- 2544520 TI - Red blood cell lithium-sodium countertransport in non-modulating essential hypertension. AB - Abnormalities in erythrocyte Li-Na countertransport have been reported in hypertensive subjects, and the available evidence favors familial aggregation and striking heritability of this marker. It is uncertain, however, whether the abnormalities are associated with hypertension per se or whether they may be concentrated in a particular subset of hypertensive subjects. In the present study, maximal rates of Li-Na countertransport were measured in red blood cells of 82 white subjects, including 37 normotensive subjects and 45 normal- or high renin hypertensive subjects previously classified as non-modulators (n = 21) or modulators (n = 24). Mean countertransport activity was significantly higher in non-modulators compared with normally modulating hypertensive or normotensive subjects (0.475 +/- 0.044 vs. 0.309 +/- 0.028 or 0.249 +/- 0.012 mmol/l cell x hr, respectively, p less than 0.001). Modulators did not differ significantly from normotensive subjects with regard to mean countertransport activity. Red blood cell sodium pump and Na-K-Cl cotransport were not significantly different in modulating and non-modulating hypertensive subjects. These relations remained unchanged after adjusting for age, body weight, and plasma cholesterol levels by analysis of covariance. A countertransport value exceeding 0.50 mmol/l cell x hr occurred in 40% of the non-modulators but in only one of the other subjects. In contrast , while one half of the modulators and normotensive subjects had a countertransport value less than 0.235 mmol/l cell x hr, none of the non modulators did. Thus, elevated countertransport appears to aggregate in the non modulating subset of essential hypertensive subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544521 TI - Kinin antagonist reverses converting enzyme inhibitor-stimulated vascular prostaglandin I2 synthesis. AB - Treatment with a converting enzyme inhibitor has been shown to stimulate aortic prostaglandin I2 synthesis. We studied whether converting enzyme inhibitor stimulated prostaglandin I2 synthesis might be mediated by kinins. Anesthetized male Sprague-Dawley rats were given a continuous 70-minute infusion of either saline or a kinin analogue antagonist, [DArg0-Hyp3-Thi5-DPhe7-Thi8]bradykinin, 8 micrograms/kg/min. After 10 minutes, rats were given an intravenous bolus of either vehicle or the converting enzyme inhibitor enalaprilat (30 micrograms/100 g body wt). After 70 minutes, aorta and renal cortical slices were harvested and incubated in vitro in buffer without drugs at pH 7.4, 37 degrees C for 60 minutes. The buffer was then sampled for measurement of 6-keto prostaglandin F1 alpha (an index of prostaglandin I2), prostaglandin E2, and renin release (angiotensin I generation) by radioimmunoassay. The aortic prostaglandin I2 from rats treated with converting enzyme inhibitor was significantly elevated (36.7 +/ 5.0 ng/mg dry wt/hr) compared with aorta from rats treated with either vehicle (25.6 +/- 2.2 ng/mg/hr), kinin antagonist (25.1 +/- 2.4 ng/mg/hr), or kinin antagonist plus converting enzyme inhibitor (23.0 +/- 2.0 ng/mg/hr), p less than 0.02. There were no differences in aortic prostaglandin E2, renin release, or prostaglandin E2 from renal cortical slices. Direct in vitro incubation of aorta with molar concentrations of converting enzyme inhibitor from 10(-9) to 10(-4) had no effect on prostaglandin I2. These results suggest that kinins may mediate the effect of converting enzyme inhibition on aortic prostaglandin I2 synthesis and thereby may account for part of the hemodynamic responses resulting from treatment using converting enzyme inhibitors. PMID- 2544522 TI - Cardiovascular effects of the N-methyl-D-aspartate receptor antagonist MK-801 in conscious rats. AB - Evidence from microinjection studies in anesthetized rats suggests that central excitatory amino acid pathways using N-methyl-D-aspartate receptors are involved in the regulation of the cardiovascular system. To test the hypothesis that these pathways are tonically involved in the maintenance of or the baroreceptor reflex regulation of cardiovascular function, we have examined the effects of intravenous injection of the centrally acting, noncompetitive N-methyl-D aspartate receptor antagonist (+)-5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine (MK-801), on the mean arterial pressure, heart rate, renal sympathetic nerve activity, and behavior of conscious, freely moving sham-operated and sinoaortic baroreceptor-denervated rats. Administration of MK 801 produced, within 5 minutes, dose-dependent elevations in mean arterial pressure, heart rate, and renal sympathetic nerve activity that were sustained for 0.5 to 2.5 hours. For an equivalent dose, MK-801 produced approximately twice the peak changes in mean arterial pressure and heart rate in the sinoaortic baroreceptor-denervated rats than in the sham-operated rats. Pretreatment results were as follows: 1) The ganglion blocker chlorisondamine markedly attenuated the hypertension and tachycardia in the sham-operated and sinoaortic baroreceptor denervated rats, 2) pretreatment with the alpha 1-adrenergic receptor antagonist prazosin virtually abolished the hypertension, and 3) the beta 1-adrenergic receptor antagonist atenolol markedly reduced the tachycardia. MK-801 also produced stereotypic behaviors and ataxia in the sham-operated and sinoaortic baroreceptor-denervated rats; however, qualitatively and quantitatively similar changes in behavior were induced in the latter by doses approximately five time lower than required in sham operated rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544523 TI - Long-term improvement in renal function after short-term strict blood pressure control in hypertensive nephrosclerosis. AB - Seventy-nine hypertensive nephrosclerosis patients entered a prospective randomized single-blind study to 1) establish the pattern of decay of renal function in this population and the variability therein and 2) to determine if strict diastolic blood pressure (DBP) control (less than or equal to 80 mm Hg) is more effective than conventional levels (90-95 mm Hg) in conserving renal function. Because of unexpected significant improvement in renal function in patients from both groups, which changed the perspectives on the course of this disease as described herein, this report is being published before completion of the trial. The selection criteria were 1) serum creatinine concentration of 1.6 7.0 mg/dl, 2) glomerular filtration rate of less than 70 ml/min/1.73 m2, and 3) absence of diseases (other than hypertension) known to destroy renal function. Renal function was assessed by glomerular filtration rate [( 125I]iothalamate clearance) and serum creatinine concentration. Before randomization, DBP was aggressively treated to reduce it to less than 80 mm Hg. Twenty-two subjects (14 in the strict DBP control group and eight in the conventional DBP control group) have been enrolled in the study for 36 months. In contrast to results from previous studies in humans and rats, renal function improved in both patient groups. Thus, irrevocable progression of renal damage after onset of renal failure from high blood pressure does not necessarily occur, and in fact, long term improvement of renal function resulted from the effects of the study itself.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544524 TI - Agonist-induced vascular tone. AB - The cellular mechanisms underlying the agonist-induced sustained contraction of the vascular smooth muscle are reviewed in the light of the use of Ca2+ and the change of Ca2+ sensitivity of the contractile apparatus. It is generally accepted that the main trigger for contraction of vascular smooth muscle is the elevation of intracellular Ca2+ concentration. However, the measurement of intracellular Ca2+ concentration during the sustained phase of agonist-induced contraction is reported to be lower than that of high K+ stimulation or the value obtained by the experiments with chemically skinned smooth muscle preparations. These observations indicate that a second regulatory system may exist. One possible mechanism is the effectiveness of Ca2+ use. Agonist-induced Ca2+ influx may be more effective in raising the intracellular Ca2+ in the bulk of the cytoplasm than is Ca2+ entry induced by depolarization by the inhibition of a putative sarcoplasmic reticulum buffer barrier. Another possibility is the change of Ca2+ sensitivity of the contractile apparatus. Although the survey of the recent literature concerning the phorbol ester-induced vasoconstriction tends to support a role for protein kinase C in the change of Ca2+ sensitivity of the contractile proteins, it fails to establish a clear link between receptors, protein kinase C, and myofilaments. By using new methods for permeabilizing smooth muscle fibers, which retain the function of receptors and signal transduction systems, we now provide direct evidence that the activation of G protein by norepinephrine or guanosine 5'-0-(3-triphosphate) (GTP-gamma-S), nonhydrolyzable GTP analogue, enhances myofilament sensitivity to Ca2+. PMID- 2544525 TI - Characterization of intrarenal arterial adrenergic receptors in renovascular hypertension. AB - alpha-Adrenergic receptor subtypes were investigated using [3H]prazosin, an alpha 1 selective antagonist, and the alpha 2 selective antagonist [3H]rauwolscine in a smooth muscle plasma membrane enriched microsomal fraction prepared from rabbit intrarenal arterial vasculature. Both radioligands displayed single components on Scatchard analysis. The specific binding of [3H]prazosin was of high affinity (0.54 +/- 0.04 nM) with a maximum binding capacity (Bmax) of 212 +/- 15 fmol/mg protein. The maximum number of [3H]rauwolscine binding sites was 64 +/- 4 fmol/mg of protein with a dissociation constant (Kd) of 5.60 +2- 0.27 nM. Binding of both radioligands was rapid, saturable, and specific. alpha 1- and alpha 2-adrenergic receptors in the intrarenal arterial membrane preparation were also characterized at 2-, 4-6-, and 10-12-week intervals during the course of development and maintenance of chronic two-kidney, one clip (2K1C) Goldblatt hypertension and in age-matched sham-operated normotensive control rabbits. The alpha 1-adrenergic receptor affinity for [3H]prazosin binding in hypertensive rabbits was significantly increased in the stenotic, but not contralateral, kidney at 2 weeks; however, at 6 weeks the receptor affinity of both kidneys was significantly increased compared with those of the normotensive control group. No difference in alpha 1-adrenergic receptor affinity was seen at 12 weeks, and there were no changes in Bmax at any of the weekly intervals. Neither the Kd, nor Bmax, for [3H]rauwolscine in either kidney showed a significant difference between hypertensive rabbits and normotensive control rabbits. These studies demonstrate the existence in the rabbit intrarenal arterial vasculature of binding sites with alpha 1- and alpha 2-adrenergic receptor specificity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544526 TI - Effect of 8-bromo-cyclic guanosine monophosphate on intracellular pH and calcium in vascular smooth muscle. AB - Endothelium-dependent vasodilators, nitrates, and atrial natriuretic factor relax blood vessels by increasing vascular cyclic guanosine monophosphate (cGMP). The mechanisms by which cGMP relaxes vascular smooth muscle (VSM) are not known. Since contraction of VSM is associated with increased intracellular calcium and pH, we hypothesized that cGMP may decrease vascular tone by lowering ionized, intracellular calcium [( Ca2+]i) and pH. We used microfluorometry to measure cGMP induced changes in intracellular calcium and pH of cultured A7r5 VSM cells after stimulation with contractile agonists. A cGMP analogue, 8-Br-cGMP, blocked vasopressin- but not thrombin-stimulated increases in [Ca2+]i. High extracellular potassium concentrations [( K+]) increased [Ca2+]i, but the attenuation of [Ca2+]i by 8-Br-cGMP was not statistically significant. 8-Br-cGMP also attenuated vasopressin- but not thrombin-stimulated alkalinization of VSM cells. cGMP may decrease vascular tone by decreasing [Ca2+]i and pH, but these changes are dependent on the contractile agonist studied. PMID- 2544527 TI - Urinary sodium pump inhibitor raises cytosolic free calcium concentration in rat aorta. AB - We were able to purify two distinct sodium pump inhibitors to homogeneity from human urine based on [3H]ouabain-displacing activity from intact human erythrocytes. The polar and less polar compounds were eluted off the C18 reverse phase column with 18% and 31% acetonitrile, respectively. The polar compound cross-reacted very weakly with specific antidigoxin antibody and lacked a characteristic ultraviolet absorption peak between 190 and 300 nm. The less polar compound showed a prominent digoxinlike immunoreactivity and had an ultraviolet spectrum similar to that of digoxin. We examined the effects of these compounds on cytosolic free calcium concentration in cultured rat vascular smooth muscle cells (A10 cells) using the fluorescent calcium chelator fura-2. Only the polar ouabain-displacing compound caused a significant increase, from 108 +/- 7 to 162 +/- 8 nM (n = 6, p less than 0.01), in cytosolic free calcium concentration in A10 cells. The rise in cytosolic free calcium concentration induced by the polar ouabain-displacing compound tended to be slower in onset and more sustained than that induced by arginine vasopressin. In contrast, ouabain and bufalin had no appreciable effects on cytosolic free calcium concentration in A10 cells. These results suggest that the polar ouabain-displacing compound we isolated from human urine may possess a vasoactive property and may play an important role in the modulation of vascular tone. PMID- 2544528 TI - Beta-receptor properties in soleus muscles from spontaneously hypertensive rats. AB - We have compared the properties of beta-adrenergic receptors in slow-twitch, oxidative skeletal muscles (soleus) from spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats at three different ages. The investigation was based on the hypothesis that the increase in Na+ content and decrease in fatigue resistance observed previously in the soleus of SHR might be the result of a down regulation of muscle beta-receptors. Activation of beta-adrenergic receptors in skeletal muscle stimulates sarcolemmal sodium-potassium adenosine triphosphatase, which produces an efflux of Na+ and an influx of K+. Receptor down-regulation would be expected to reduce hormonal stimulation of Na+ pump activity, particularly during exercise. The results of receptor binding studies, however, and an investigation of cyclic adenosine monophosphate (cAMP) production in response to applied epinephrine indicated that there were no significant differences in receptor properties in the soleus muscles of SHR and WKY rats. Receptor number and affinity were the same in the two strains, and the rate, magnitude, and duration of the increase in cAMP in response to 10(-6) M epinephrine were also similar. beta-Adrenergic receptor down-regulation does not appear to be a generalized phenomenon in tissues of SHR, despite the appearance of other physiological changes in the tissue. PMID- 2544529 TI - Potentiation of renal effects of atrial natriuretic factor-(99-126) by SQ 29,072. AB - Depressor and renal activities of atrial natriuretic factor-(99-126) were determined in conscious, unrestrained spontaneously hypertensive rats treated with a neutral endopeptidase inhibitor, SQ 29,072 (7-[[2-(mercaptomethyl)-1-oxo-3 phenylpropyl]amino]heptanoic acid). SQ 29,072 (100 mumol/kg i.v.) prolonged the transient depressor effects of the peptide for as long as 2 hours. During the first hour after 3, 10, and 30 nmol/kg atrial natriuretic factor, urinary excretion of cyclic 3'5' guanosine monophosphate was significantly increased by 9.2 +/- 3.4, 13.0 +/- 2.2, and 12.7 +/- 4.2 nmol/kg/hr, respectively, in vehicle treated rats and by 26.9 +/- 7.9, 52.1 +/- 11.1, and 46.4 +/- 12.2 nmol/kg/hr, respectively, in rats given 100 mumol/kg SQ 29,072. During the first hour after 3 and 10 nmol/kg atrial natriuretic factor-(99-126), the sodium loss was 161 +/- 56 and 139 +/- 42 mueq/kg/hr in vehicle-treated rats and was significantly greater (694 +/- 316 and 1,038 +/- 135 mueq/kg/hr) in rats given 100 mumol/kg SQ 29,072. After administration of 3, 10, and 30 mumol/kg SQ 29,072, the area over the curves of the depressor responses to 3 nmol/kg of the peptide increased from 297 +/- 70 to 306 +/- 108, 440 +/- 143, and 669 +/- 186 mm Hg.min, respectively, while the concurrent natriuretic responses rose from 161 +/- 56 to 250 +/- 88, 332 +/- 142, 464 +/- 164, and 694 +/- 316 mueq/kg/hr. In summary, the neutral endopeptidase inhibitor SQ 29,072 increased the magnitudes and especially the durations of the depressor, natriuretic, and cyclic guanosine monophosphate responses to exogenous atrial natriuretic factor-(99-126) in conscious spontaneously hypertensive rats, presumably by inhibition of degradation of atrial natriuretic factor in vivo. In conclusion, neutral endopeptidase inhibition offers an important new technique for enhancement and prolongation of the biological lifetime of atrial natriuretic factor. PMID- 2544530 TI - Immunopharmacological studies on Picrorhiza kurroa Royle-ex-Benth. Part IV: Cellular mechanisms of anti-inflammatory action. AB - In this work abroagation of anti-inflammatory effect of Picrorhiza kurroa extract (PK) by beta-adrenergic blockade was confirmed, which suggests alteration in cell surface biology by PK treatment. Blockade of protein synthesis by cycloheximide pretreatment reduced PK effect, suggesting protein mediation. Metabolic inhibitor dinitrophenol inhibited inflammatory cedema equally in control and PK treated animals, and masking of PK effect was concluded. Discriminations of anti inflammatory mechanism(s) of PK and the latter two cytotoxic agents was inferred from these observations and from existing knowledge. Selective PK influence on membrane linked activation events in inflammatory effector cells could be the basis of anti-inflammatory and perhaps other biological activities reported with the herb. PMID- 2544533 TI - Visual compatibility of enalaprilat with commonly used critical care medications during simulated Y-site injection. AB - Physical incompatibility studies between an intravenous angiotensin-converting enzyme inhibitor, enalaprilat, and potentially coadministrable ICU medications have been performed. Forty-one medications and four solutions were evaluated. The medications were anesthetic/narcotics, antibiotics, an anticoagulant, a bronchodilator, electrolyte solutions, fluids, H-2 receptor blocking agents, steroids and vasoactive agents. There was no turbidity, precipitation, or color change caused by the admixing of enalaprilat with any of the 45 agents studied. This study did not take bioavailability into consideration. PMID- 2544532 TI - Human parvovirus B19 infection associated with vascular purpura and vasculitis. PMID- 2544531 TI - Fluconazole in the treatment of fungal infections associated with AIDS. PMID- 2544534 TI - Macrophage activation and host augmentation against Sendai or herpes simplex virus (HSV) infections with synthetic polypeptides in mice. AB - Poly-L-Lys (mean mol. wt; 12,000), poly-L-Arg (5000) and poly-L-Orn were found to activate peritoneal macrophages effectively in vivo in 14 synthetic homo polypeptides. The ability of sequential poly(L-Arg-L-X) (5000) to activate macrophages was less than that of poly-L-Arg. Neither (L-Arg)12 nor (L-Arg)6 by themselves activated macrophages, but poly-D-Arg (5000) did, as also did poly-L Arg; this suggests that the polycationic character of poly-L-Arg plays a role in the activation of macrophages. The intranasal administration of poly-L-Lys, -L Arg, -L-Orn, -D-Arg, all of which activated macrophages, augmented host resistance against Sendai virus infection in mice. The protection afforded by poly-L-Arg seemed to depend on its mol. wt: the order of protection was poly-L Arg greater than (L-Arg)12 greater than (L-Arg)6. The intranasal administration of poly-L-Arg 3 days before the infection was effective, while that 1 day before infection was not. There was no difference between the groups in the titer of interferon produced by the infection of Sendai virus given with poly-L-Arg either 3 days before or 1 day before the infection. The administration of poly-L-Arg 3 days before the infection caused a significant decrease of the virus titer in the lung 6 days after the infection when compared with the control or the mice given 1 day before. The intravenous administration of 2-chloroadenosine (2-Cl-Ade), which is a selective inhibitor of macrophages, into the mice which had received poly-L-Arg intranasally 3 days before the infection caused a significant decrease in the survival rate of the mice, indicating that the macrophages activated with poly-L-Arg are likely to be an important element in affording the protection. Subcutaneous administration of poly-L-Arg revealed protective activity against systematic infection with herpes virus-type 1. PMID- 2544535 TI - Water exchange through the erythrocyte membrane. PMID- 2544536 TI - [Painless ulcer of the external ear]. PMID- 2544537 TI - [Hemolytic anemia in a 32-year-old patient with metastatic breast cancer]. PMID- 2544538 TI - Membrane damage in acute brain trauma. AB - Among a number of biochemical disturbances occurring in the acute phase of brain insults, the destruction of membrane phospholipids and its consequences on the function of membrane-bound proteins is likely to be one of the most important. In the cryogenic type of injury which is classically considered as a relevant animal model of brain contusive lesions in human traumatology, the initial attack of membranes could consist in a peroxidative damage triggered by blood ferrous compounds. This in turn would lead to an activation of phospholipase A2. As a consequence of phospholipid disruption a number of enzymes involved in energy production within the mitochondria are severely impaired. Nevertheless, the level of available ATP within the cell remains normal and even higher than normal. This paradoxical findings suggests that energy utilization is even more lowered than energy production. In fact, the Na+-K+-ATPase activity which normally utilizes approximately 70% of the total amount of cellular energy is severely reduced. We assume that Na+-K+-ATPase impairment is directly responsible for the retention of intracellular Na+ accompanied by osmotically driven water, though admittedly other biochemical disturbances, including tissue acidosis and liberation of excitatory amino-acids, would contribute to the same result. Lastly, a striking feature of these biochemical events is the early activation of those enzymes necessary for phospholipid resynthesis. This should mean that repair processes are at work immediately after the insult allowing resumption of Na+-K+-ATPase function, clearing up of brain edema and restoration of cation exchanges essential for brain work. PMID- 2544539 TI - Chronic infection with Epstein-Barr virus, Chlamydia and hepatitis A virus, terminating in cirrhosis and nasopharyngeal carcinoma. AB - We describe a 27-year-old woman who presented with infertility, and over the subsequent 7 years developed chronic Epstein-Barr virus infection, hepatitis A, cirrhosis and nasopharyngeal carcinoma. Serological evidence of chronic infection with Chlamydia, Epstein-Barr virus and hepatitis A virus was documented. PMID- 2544540 TI - Breast tuberculosis and carcinoma. PMID- 2544541 TI - [Non-familial ulcero-mutilating acro-osteopathy of the feet]. PMID- 2544542 TI - Characterization of a new alpha-thalassemia-1 deletion in a Spanish family. AB - A new type of alpha-thalassemia-1 was characterized in one Spanish patient with Hb H disease and in her mother. The restriction map of this deletion suggests that the deletion of 22 kb has occurred on a chromosome carrying a zeta-globin triplication. The resulting chromosome lacks the alpha 2- and alpha 1-globin genes, the psi alpha 2- and psi alpha 1-globin genes, and one of the three zeta globin genes, while the other two zeta-globin genes and the theta 1-globin gene have been retained. PMID- 2544543 TI - Krukenberg's tumor in the young population. AB - Krukenberg's tumor secondary to gastric adenocarcinoma was identified in a 20 year-old woman. The patient died ten weeks after surgery, which included total hysterectomy, bilateral salpingo-oophorectomy, tumor debulking, and subtotal gastrectomy. Her unexpected advanced disease and fulminant course led the authors to review the literature for other reports of Krukenberg's tumor in the young person. However, the age category was not studied specifically in most series, and only a few cases involving patients of very young age were detailed. The present case appears to be only the fifth involving a patient aged 20 years. PMID- 2544544 TI - Lanthiopeptin, a new peptide antibiotic. Production, isolation and properties of lanthiopeptin. AB - A strain of Streptoverticillium cinnamoneum produced a peptide antibiotic named lanthiopeptin, which contained four unusual amino acids, erythro-beta hydroxyaspartic acid, mesolanthionine, threo-beta-methyllanthionine and lysinoalanine. Lanthiopeptin showed antiviral activity against herpes simplex virus type 1 KOS strain infection in Vero cells by cytopathic effect reduction assay. The structure of lanthiopeptin is similar to that of ancovenin. PMID- 2544545 TI - Establishment of a murine malignant fibrous histiocytoma cell line (DIM-L) with lung metastatic potential. PMID- 2544546 TI - A new human cholangiocellular carcinoma cell line (HuCC-T1) producing carbohydrate antigen 19/9 in serum-free medium. AB - A human cholangiocellular carcinoma cell line, HuCC-T1, was established in vitro from the malignant cells of ascites of a 56-yr-old patient. Histologic findings of the primary liver tumor revealed a moderately differentiated adenocarcinoma. Tumor cells from the ascites have been cultured with RPMI 1640 medium containing 0.2% lactalbumin hydrolysate and the cultured cells grew as monolayers with a population doubling time of 74 h during exponential growth at Passage 25. They had an epithelial-like morphology and were positive for mucine staining. Ultrastructural studies revealed the presence of microvilli on the cell surface and poorly developed organelles in the cytoplasm. The HuCC-T1 cell was tumorigenic in nude mice. The number of chromosomes in HuCC-T1 ranged from 61 to 80. These human cholangiocellular carcinoma cells in serum-free medium secreted several tumor markers, including carbohydrate antigen 19/9, carbohydrate antigen 125, carcinoembryonic antigen, and tissue polypeptide antigen. The carbohydrate antigen 19/9 secretion level of HuCC-T1 cells cultured in RPMI 1640 medium with 1% fetal bovine serum was sixfold higher than that with 0.2% lactalbumin hydrolysate. These findings suggest that HuCC-T1 will provide useful information to clarify the mechanism of tumor marker secretion and tumor cell growth in the human cholangiocellular carcinoma. PMID- 2544547 TI - Clonal populations of the mouse mammary cell line, COMMA-D, which retain capability of morphogenesis in vivo. AB - Clonal populations were isolated from the mouse mammary cell line, COMMA-D, by transfection with a dominant-selectable gene, pSV2Neo, which confers resistance to the antibiotic, G418. Seven of twenty-four clones isolated retained the ability of the parental line to repopulate cleared mammary fat pads in vivo as ductal-alveolar hyperplasias. Two sublines designated CDNR2 and CDNR4 retained hyperplastic growth potential after multiple passages in vitro with low incidence of tumor formation. A third subpopulation, CDNR1, contained a single integration site for the pSV2Neo plasmid indicating a bonafide clonal origin for this subline. CDNR1 cells displayed heterogeneous growth phenotypes in vivo including hyperplasia, adenocarcinoma, and bone formation. Functional differentiation of CDNR1 cells organized as alveolarlike structures in vivo or on floating collagen gels in vitro was observed as determined by immunoperoxidase staining for the milk-specific protein, casein. Overall, the results indicate that a subset of cells from the COMMA-D cell line may be functionally analogous to stem cells existing in the mammary gland. PMID- 2544548 TI - Tn4399, a conjugal mobilizing transposon of Bacteroides fragilis. AB - Conjugal transposons play an important role in the dissemination of antibiotic resistance determinants in the streptococci and have been postulated to exist in Bacteroides fragilis. To investigate the presence of conjugal transposons in B. fragilis, we employed a Tra- derivative of the transfer factor pBFTM10 contained in the chimeric plasmid pGAT400 delta BglII. We attempted to restore transferability to this plasmid from a series of transconjugants generated by crossing B. fragilis TMP230 containing the TET transfer factor with B. fragilis TM4000, a standard recipient. Transconjugant TM4.2321 transferred pGAT400 delta BglII to Escherichia coli HB101 at almost the same frequency as did the Tra+ parental plasmid, pGAT400. Analysis of the transferred plasmids revealed the presence of 9.6 kilobases of additional DNA in every case but at different positions in independent isolates. The presence of this DNA, designated Tn4399, allowed the pGAT400 delta BglII derivatives to retransfer from the TM4000 background to B. fragilis or E. coli recipients. DNA hybridization studies demonstrated the presence of one copy of Tn4399 in TMP230 and three copies at new sites in TM4.2321. Tn4399 is a new B. fragilis transposon with unique transfer properties that may play a role in the dissemination of drug resistance genes. It differs from previously described conjugal transposons by its ability to mobilize nonconjugal plasmids in cis. PMID- 2544549 TI - Molecular analysis of the Escherichia coli recO gene. AB - The plasmid pLC7-47, which contains lep, rnc, and era, was found to complement the UV-sensitive and recombination-deficient phenotypes caused by the recO1504::Tn5 mutation. Southern blotting analysis demonstrated that pLC7-47 contained a segment of Escherichia coli DNA that covered the region of the E. coli chromosome containing the recO1504::Tn5 mutation. A combination of deletion mapping and insertional mutagenesis localized the recO-complementing region to an approximately 1-kilobase region of a 1.6-kilobase BamHI fragment. The DNA sequence of the 1.6-kilobase BamHI fragment was determined and contained part of era and a 726-base-pair recO open reading frame. The recO open reading frame contained three possible translation start codons and could potentially encode a polypeptide of Mr 26,000. Computer analysis indicated that the putative RecO protein had suboptimal codon usage and did not show significant homology with previously identified proteins whose sequences were present in protein data bases. A combination of primary sequence analysis and secondary structure predictions suggested that recO contains a mononucleotide-binding fold. PMID- 2544550 TI - Control of mucoidy in Pseudomonas aeruginosa: transcriptional regulation of algR and identification of the second regulatory gene, algQ. AB - A new alginate regulatory gene, algQ, was identified in a chromosomal region which, when tandemly amplified, induces mucoidy in Pseudomonas aeruginosa. The algQ gene was found closely linked to the previously identified algR gene. Both algQ and algR were required for transcription of the key alginate biosynthetic gene, algD. In addition, expression of the algR gene was studied. The algR promoter was mapped by S1 nuclease and reverse transcription and found to be activated in mucoid cells. However, even in nonmucoid cells, transcription of algR was detectable at an approximately 50-fold-lower level, as opposed to the algD promoter, which was silent in the nonmucoid background. Transcription of both promoters was studied by using algR- and algD-specific oligonucleotides and total cellular RNA from fresh cystic fibrosis isolates of mucoid P. aeruginosa and their nonmucoid revertants. Identical patterns of activity were found in all strains: in mucoid cells, both algR and algD were activated. This finding indicated that common mechanisms were involved in the regulation of alginate gene expression. However, when the algR gene was cloned behind the tac promoter on a broad-host-range-controlled expression vector, induction of transcription with isopropropyl-beta-D-thiogalactopyranoside (IPTG) caused the appearance of a nonmucoid phenotype in previously mucoid cells. This effect was transient, since removal of the inducer (IPTG) made cells mucoid again. Since the algR gene product is homologous to transcriptional regulators from a class of environmentally responsive systems (known to have a second, sensory component), the algQ gene could be a candidate for the sensory component of the alginate system. PMID- 2544551 TI - Multiple defects in Escherichia coli mutants lacking HU protein. AB - The HU protein isolated from Escherichia coli, composed of two partially homologous subunits, alpha and beta, shares some of the properties of eucaryotic histones and is a major constituent of the bacterial nucleoid. We report here the construction of double mutants totally lacking both subunits of HU protein. These mutants exhibited poor growth and a perturbation of cell division, resulting in the formation of anucleate cells. In the absence of HU, phage Mu was unable to grow, to lysogenize, or to carry out transposition. PMID- 2544552 TI - dsg, a gene required for cell-cell interaction early in Myxococcus development. AB - dsg mutants of Myxococcus xanthus are conditionally defective in fruiting body development, including sporulation. Unable to develop on their own, these mutants can assemble fruiting bodies with spores if they are mixed with wild-type cells. To elucidate the developmental defect in dsg mutants by close comparison with wild type, such mutants have been backcrossed by transduction, using a closely linked insertion of transposon Tn5 for selection. Backcrossed dsg mutants form aggregates that are larger, less compact, and less symmetrical than dsg+ fruiting bodies. Also, the starvation-induced sporulation in dsg aggregates is delayed and reduced. However, dsg mutants can be induced by glycerol or dimethyl sulfoxide to sporulate at levels approaching those of wild type. dsg mutants may thus have a primary defect early in development which diminishes their capacity to aggregate and which indirectly decreases the number of fruiting body spores. The linked insertion of Tn5 also facilitated cloning the dsg gene. The cloned dsg+ allele was shown to be dominant to both the dsg-429 and dsg-439 alleles, and both mutant alleles were shown to belong to the same genetic complementation group. Subcloning of restriction fragments, deletions, and insertions of transposon Tn5 agree in locating the dsg gene to an 850-base-pair segment of the cloned region. PMID- 2544553 TI - dsg, a gene required for Myxococcus development, is necessary for cell viability. AB - Previous work identified the dsg gene as necessary for cell-cell interaction in Myxococcus xanthus. Point mutations of this gene, such as dsg-439, are viable, but insertions of Tn5 within the dsg gene (dsg::Tn5) are lethal. Partial diploids, dsg::Tn5/dsg+ or dsg::Tn5/dsg-429 or dsg::Tn5/dsg-439, are also viable, showing that the lethal effect of the haploid insertions is due to loss of function. Thus the evidence implies that the dsg gene is essential for viability as well as development, but its essential quality differs between growth and development because dsg-429 and dsg-439 mutants grow normally, but are unable to develop. PMID- 2544554 TI - DnaA protein overproduction abolishes cell cycle specificity of DNA replication from oriC in Escherichia coli. AB - Initiation of DNA replication from oriC in Escherichia coli takes place at a specific time in the cell division cycle, whether the origin is located on a chromosome or a minichromosome, and requires participation of the product of the dnaA gene. The effects of overproduction of DnaA protein on the cell cycle specificity of the initiation event were determined by using minichromosome replication as the assay system. DnaA protein was overproduced by inducing the expression of plasmid-encoded dnaA genes under control of either the ptac or lambda pL promoter. Induction of DnaA protein synthesis caused a burst of minichromosome replication in cells at all ages in the division cycle. The magnitude of the burst was consistent with the initiation of one round of replication per minichromosome in all cells. The replication burst was followed by a period of reduced minichromosome replication, with the reduction being greater at 30 than at 41 degrees C. The results support the idea that the DnaA protein participates in oriC replication at a stage that is limiting for initiation. Excess DnaA protein enabled all cells to achieve the state required for initiation of DNA polymerization by either effecting or overriding the normal limiting process. PMID- 2544555 TI - Transposon mutagenesis, characterization, and cloning of transformation genes of Haemophilus influenzae Rd. AB - A plasmid library of PstI fragments of Haemophilus influenzae Rd genomic DNA was mutagenized in Escherichia coli with mini-Tn10kan. The mutagenized PstI fragments were introduced by transformation into the H. influenzae chromosome, and kanamycin-resistant transformants were screened for the transformation-deficient phenotype by a cyclic AMP-DNA plate method. Fifty-four mutant strains containing 24 unique insertions that mapped to 10 different PstI fragments were isolated. Strains carrying unique insertions were tested individually for DNA uptake, transformation efficiency, UV sensitivity, and growth rate. The transformation frequencies of these mutants were decreased by factors of 10(-2) to 10(-6). Five of the mutants had normal competence-induced DNA uptake, and the rest were variably deficient in competence development. Three strains were moderately UV sensitive. All strains but one had doubling times within 50% of that of the wild type. Mutated genes were cloned into an H. influenzae-E. coli shuttle vector, and wild-type loci were recovered by in vivo recombinational exchange. Hybridization of these clones to SmaI genomic fragments separated in pulsed-field gels showed that these insertions were not clustered in a particular region of the chromosome. PMID- 2544556 TI - Segregation of relaxed replicated dimers when DNA ligase and DNA polymerase I are limited during oriC-specific DNA replication. AB - An in vitro Escherichia coli oriC-specific DNA replication system was used to investigate the DNA replication pathways of oriC plasmids. When this system was perturbed by the DNA ligase inhibitor nicotinamide mononucleotide (NMN), alterations occurred in the initiation of DNA synthesis and processing of intermediates and DNA products. Addition of high concentrations of NMN soon after initiation resulted in the accumulation of open circular dimers (OC-OC). These dimers were decatenated to open circular monomers (form II or OC), which were then processed to closed circular supercoiled monomers (form I or CC) products. After a delay, limited ligation of the interlinked dimers (OC-OC to CC-OC and CC CC) also occurred. Similar results were obtained with replication protein extracts from polA mutants. The presence of NMN before any initiation events took place prolonged the existence of nicked template DNA and promoted, without a lag period, limited incorporation into form II molecules. This DNA synthesis was nonspecific with respect to oriC, as judged by DnaA protein dependence, and presumably occurred at nicks in the template DNA. These results are consistent with oriC-specific initiation requiring closed supercoiled molecules dependent on DNA ligase activity. The results also show that decatenation of dimers occurs readily on nicked dimer and represents an efficient pathway for processing replication intermediates in vitro. PMID- 2544557 TI - Identification of a second gene involved in global regulation of fumarate reductase and other nitrate-controlled genes for anaerobic respiration in Escherichia coli. AB - Fumarate reductase catalyzes the final step of anaerobic electron transport in Escherichia coli when fumarate is used as a terminal electron acceptor. Transcription of the fumarate reductase operon (frdABCD) was repressed when cells were grown in the presence of either of the preferred terminal electron acceptors, oxygen or nitrate, and was stimulated modestly by fumarate. We have previously identified a locus called frdR which pleiotropically affects nitrate repression of fumarate reductase, trimethylamine N-oxide reductase, and alcohol dehydrogenase gene expression and nitrate induction of nitrate reductase expression (L. V. Kalman and R. P. Gunsalus, J. Bacteriol. 170:623-629, 1988). Transformation of various frdR mutants with plasmids identified two complementation groups, indicating that the frdR locus is composed of two genes. One class of mutants was not completely restored to wild-type frdA-lacZ expression or nitrate reductase induction when complemented with multicopy narX+ plasmids, whereas low-copy narX+ plasmid-containing strains were. A second class of frdR mutants was identified and shown to correspond to a previously described gene, narL (frdR2). Complementation of these strains with multicopy narL+ plasmids resulted in superrepression of frdA-lacZ expression and moderate elevation of nitrate reductase expression. Multicopy plasmids containing both narL+ and narX+ or only narL+ were able to complement narL mutants, whereas narX+ plasmids complemented narX mutants only when present in a copy number approximately equal to that of narL. Both narL and narX mutants retained normal oxygen control of frdA-lacZ expression. Both types of mutants are pleiotropic, as evidenced by derepressed levels of the fumarate reductase and trimethylamine N oxide reductase enzymes and by defective induction of nitrate reductase when cells were grown in the presence of nitrate. These results indicate that both the narL and narX gene products must be present in a defined ratio in the cell. We conclude that these proteins interact to effect normal nitrate control of the anaerobic electron transport-associated operons. From these studies, we propose that narX encodes a nitrate sensor protein while narL encodes a DNA-binding regulatory protein which together function in a manner analogous to other two component regulatory systems. PMID- 2544558 TI - Oxygen, nitrate, and molybdenum regulation of dmsABC gene expression in Escherichia coli. AB - Escherichia coli can respire anaerobically using either trimethylamine-N-oxide (TMAO) or dimethyl sulfoxide (DMSO) as the terminal electron acceptor for oxidative phosphorylation. To determine whether the regulation of the dmsABC genes, which encode a membrane-associated TMAO/DMSO reductase, are transcriptionally controlled in response to the availability of alternate electron acceptors, we constructed an operon fusion between the dmsA gene, along with its associated regulatory region, and lacZ+. Expression of dmsA'-lacZ was stimulated 65-fold by anaerobiosis versus aerobiosis, while nitrate caused a 12 fold repression. Its expression, however, was unaffected by the presence of the alternate electron acceptors DMSO, TMAO, and fumarate. Anaerobic induction of dmsA'-lacZ was defective in an fnr mutant, thus establishing that Fnr is responsible for anaerobic activation of dmsABC. Repression of dmsA'-lacZ expression by nitrate was independent of oxygen and was shown to be mediated by the products of two genes, narL (frdR2) and narX. dmsA'-lacZ expression was also altered in chlD strains that are defective in molybdenum transport but not in chlA and chlE strains that are defective in molybdopterin cofactor biosynthesis, thus establishing that the molybdenum ion but not the ability to form a functional cofactor is required for regulation. Molybdenum was required both for complete induction of dmsA'-lacZ expression during anaerobic growth and for complete repression of dmsA'-lacZ by nitrate. Additionally, expression of dmsABC varied depending on the carbon source. Expression was highest when cells were grown on sorbitol. PMID- 2544559 TI - Identification and sequence analysis of the Bacillus subtilis W23 xylR gene and xyl operator. AB - The xyl operator of Bacillus subtilis W23 was identified by deletion analysis of the xyl regulatory region as a 25-base-pair (bp) sequence located 10 bp downstream from the xyl promoter. The outer 10 bp of the xyl operator exhibit perfect palindromic symmetry, while 5 central bp are nonpalindromic. It was demonstrated that the penultimate base pair near the end of this sequence is important for repressor binding. The location of the xylR gene encoding the repressor was determined by its ability to mediate xylose-dependent repression of a xyl-cat fusion on a multicopy plasmid. The nucleotide sequence of 1,355 bp from this DNA was analyzed and contains an open reading frame with a coding capacity for 384 amino acids leading to a protein with a calculated molecular weight of 42,270. A mutant with a deletion in this reading frame showed no repression of the xyl-cat fusion. The coding sequence is preceded by a suitable ribosome binding sequence and uses GTG as a start codon and TAA as a stop codon. The relationship of these results to corresponding data obtained from B. subtilis 168 is discussed. PMID- 2544560 TI - Impaired expression of certain prereplicative bacteriophage T4 genes explains impaired T4 DNA synthesis in Escherichia coli rho (nusD) mutants. AB - The Escherichia coli rho 026 mutation that alters the transcription termination protein Rho prevents growth of wild-type bacteriophage T4. Among the consequences of this mutation are delayed and reduced T4 DNA replication. We show that these defects can be explained by defective synthesis of certain T4 replication recombination proteins. Expression of T4 gene 41 (DNA helicase/primase) is drastically reduced, and expression of T4 genes 43 (DNA polymerase), 30 (DNA ligase), 46 (recombination nuclease), and probably 44 (DNA polymerase-associated ATPase) is reduced to a lesser extent. The compensating T4 mutation goF1 partially restores the synthesis of these proteins and, concomitantly, the synthesis of T4 DNA in the E. coli rho mutant. From analyzing DNA synthesis in wild-type and various multiply mutant T4 strains, we infer that defective or reduced synthesis of these proteins in rho 026-infected cells has several major effects on DNA replication. It impairs lagging-strand synthesis during the primary mode of DNA replication; it delays and depresses recombination-dependent (secondary mode) initiation; and it inhibits the use of tertiary origins. All three T4 genes whose expression is reduced in rho 026 cells and whose upstream sequences are known have a palindrome containing a CUUCGG sequence between the promoter(s) and ribosome-binding site. We speculate that these palindromes might be important for factor-dependent transcription termination-antitermination during normal T4 development. Our results are consistent with previous proposals that the altered Rho factor of rho 026 may cause excessive termination because the transcription complex does not interact normally with a T4 antiterminator encoded by the wild-type goF gene and that the T4 goF1 mutation restores this interaction. PMID- 2544562 TI - Mini-D3112 bacteriophage transposable elements for genetic analysis of Pseudomonas aeruginosa. AB - Small bacteriophage D3112 transposable elements deleted for most of the phage lytic functions while retaining the sites required for transposition and packaging were constructed to facilitate genetic studies in Pseudomonas aeruginosa. These mini-D derivatives were constructed with the terminal 1.85 kilobases (kb) of the phage left end and 1.4 kb of the phage right end and either the Tn5 kanamycin resistance or the pSC101 (pBR322) tetracycline resistance determinant. Thermally induced lysates of strains lysogenic for both a mini-D element and D3112 cts (temperature-sensitive repressor) transduced P. aeruginosa PAO recipients to drug resistance at frequencies of between 10(-4) and 10(-5)/PFU of the helper phage. As for the parent plaque-forming D3112 phage, the mini-D171 element could insert itself into many different sites in the chromosome but the frequency of insertion into particular genes varied widely. Among 1,000 insertions, none resulted in auxotrophy but 10 resulted in pigment production. Insertions were also selected in a cloning plasmid with a transduction scheme. At least eight different insertion sites were found to have been used among 10 individual insertions. Transductants harboring these mini-D elements were immune to infection by D3112, since they contained the D3112 repressor gene in the left 1.85-kb terminal fragment. Chromosomal genes were transduced in a generalized fashion 100 to 1,000 times more frequently by the mini-D-D3112 cts lysates than by the D3112 cts phage alone. Mini-D171-D3112 cts lysates also yielded some transductants that retained the drug resistance marker of the mini-D element and which were unstable for the chromosomal transduced marker. This is consistent with the miniduction properties of Mu whereby transduced genes are flanked by two mini-D elements in the same orientation. PMID- 2544561 TI - L-, P-, and M-ring proteins of the flagellar basal body of Salmonella typhimurium: gene sequences and deduced protein sequences. AB - The flgH, flgI, and fliF genes of Salmonella typhimurium encode the major proteins for the L, P, and M rings of the flagellar basal body. We have determined the sequences of these genes and the flgJ gene and examined the deduced amino acid sequences of their products. FlgH and FlgI, which are exported across the cell membrane to their destinations in the outer membrane and periplasmic space, respectively, both had typical N-terminal cleaved signal peptide sequences. FlgH is predicted to have a considerable amount of beta-sheet structure, as has been noted for other outer membrane proteins. FlgI is predicted to have an even greater amount of beta-structure. FliF, as is usual for a cytoplasmic membrane protein of a procaryote, lacked a signal peptide; it is predicted to have considerable alpha-helical structure, including an N-terminal sequence that is likely to be membrane-spanning. However, it had overall a quite hydrophilic sequence with a high charge density, especially towards its C terminus. The flgJ gene, immediately adjacent to flgI and the last gene of the flgB operon, encodes a flagellar protein of unknown function whose deduced sequence was hydrophilic and may correspond to a cytoplasmic protein. Several aspects of the DNA sequence of these genes and their surrounds suggest complex regulation of the flagellar gene system. A notable example occurs within the flgB operon, where between the end of flgG (encoding the distal rod protein of the basal body) and the start of flgH (encoding the L-ring protein) there was an unusually long noncoding region containing a potential stem-loop sequence, which could attenuate termination of transcription or stabilize part of the transcript against degradation. Another example is the interface between the flgB and flgK operons, where transcription termination of the former may occur within the coding region of the latter. PMID- 2544563 TI - In vivo cloning of Pseudomonas aeruginosa genes with mini-D3112 transposable bacteriophage. AB - The transposition properties of the Pseudomonas aeruginosa mutator bacteriophage D3112 were exploited to develop an in vivo cloning system. Mini-D replicon derivatives of D3112 were constructed by incorporating broad host range plasmid replicons between short terminal D3112 sequences. These elements were made with small replication regions from the RK2, Sa, and pVS1 plasmids and selectable genes for tetracycline, carbenicillin, kanamycin, and gentamicin resistance. Some of the mini-D replicons also contain the RK2 oriT origin-of-transfer sequence, which allows them to be mobilized by conjugation to many different species of gram-negative bacteria. These elements were used to clone DNA by preparing lysates from P. aeruginosa cells harboring an inducible D3112 cts prophage and a mini-D replicon plasmid. These lysates were used to infect sensitive P. aeruginosa recipients and select recombinant plasmids as drug-resistant transductant colonies. These transductants form a gene library from which particular clones can be selected, such as by their ability to complement specific mutations. This system was used to clone nine different genes from the PAO chromosome. The ability of this system to precisely identify a gene was demonstrated by isolating clones of the argF+ and cys-59+ genes. Restriction maps of clones of these genes, which have different amounts of flanking DNA, located the positions of these genes. The sizes of the chromosomal DNA segments from 10 individual clones examined ranged from 6 to 21 kilobases (kb), with an average of about 10 kb. This is consistent with the approximately 40-kb DNA-packaging size of the D3112 phage. PMID- 2544564 TI - Cloning, genetic characterization, and nucleotide sequence of the hemA-prfA operon of Salmonella typhimurium. AB - The first step in heme biosynthesis is the formation of 5-aminolevulinic acid (ALA). Mutations in two genes, hemA and hemL, result in auxotrophy for ALA in Salmonella typhimurium, but the roles played by these genes and the mechanism of ALA synthesis are not understood. I have cloned and sequenced the S. typhimurium hemA gene. The predicted polypeptide sequence for the HemA protein shows no similarity to known ALA synthases, and no ALA synthase activity was detected in extracts prepared from strains carrying the cloned hemA gene. Genetic analysis, DNA sequencing of amber mutations, and maxicell studies proved that the open reading frame identified in the DNA sequence encodes HemA. Another surprising finding of this study is that hemA lies directly upstream of prfA, which encodes peptide chain release factor 1 (RF-1). A hemA::Kan insertion mutation, constructed in vitro, was transferred to the chromosome and used to show that these two genes form an operon. The hemA gene ends with an amber codon, recognized by RF-1. I suggest a model for autogenous control of prfA expression by translation reinitiation. PMID- 2544565 TI - Genetic analysis of Staphylococcus aureus with Tn4001. AB - Tn4001, a 4.5-kilobase composite transposon with IS256 ends that confers resistance to gentamicin (Gmr), tobramycin, and kanamycin in Staphylococcus aureus, can transpose to diverse chromosomal sites in S. aureus. Chromosomal insertions of Tn4001 were isolated either after UV irradiation of transducing lysates carrying pII147::Tn4001 or by selection for thermoresistant Gmr isolates with strains containing thermosensitive derivatives of plasmids pI258 and pII147 carrying Tn4001. Frequent integration of the entire delivery plasmid occurred under these selective conditions in recombination-proficient hosts. When selection for thermoresistant Gmr isolates was done with these plasmids in recombination-deficient hosts, 99% or more of the Gmr isolates resulted from transposition of Tn4001 in the absence of plasmid integration. Efficient isolation of Tn4001 insertions near markers of interest and the isolation of insertional auxotrophs were achieved. Reversion frequencies of insertional auxotrophs were between 10(-6) and 10(-7) (higher than those observed with Tn551 and Tn917). About 50% of the prototrophic revertants were Gms, and these are attributed to precise excision of Tn4001. The Gmr prototrophic revertants were due to intergenic suppression. PMID- 2544566 TI - Effects of variation of inverted-repeat sequences on reactions mediated by the transposase of Tn21. AB - The frequencies of one-ended transposition and normal transposition of derivatives of Tn21 that contain mutant inverted-repeat sequences (IRs) have been measured. In general, there was a linear relationship between the log of the frequency of one-ended transposition of a mutant IR and the log of the frequency of normal transposition of an element flanked by a wild-type IR at one end and by the mutant IR at the other. This implied that one-ended and normal transposition share the rate-limiting step that determines the frequency of transposition and that both IRs are involved in the rate-limiting step in normal transposition. Surprisingly, it was found that only the outer 18 base pairs of the IR of Tn21 engaged accurately in both one-ended and normal transposition, at about 1% of the frequency of the wild-type IR. PMID- 2544567 TI - Pertussis toxin promoter sequences involved in modulation. AB - Previous analysis of the pertussis toxin (PT) promoter has shown that expression of PT requires a trans-activating factor encoded by the vir locus and a 170-base pair DNA sequence upstream from the transcription start site containing a 21-base pair direct repeat sequence crucial trans-activation (R. Gross and R. Rappuoli, Proc. Natl. Acad. Sci. USA 85:3913-3917, 1988). In this paper we extend the analysis to the modulative response to environmental stimuli. We show that modulation acts at the transcriptional level and occurs only in phase I bacteria. Modulation also requires a functional vir locus and the same promoter region of 170 base pairs. We show that, in addition to the previously identified direct repeat, even the sequences downstream from position -117 are required for trans activation and modulation and that the deletion of four cytosine residues at position -31 causes the inactivation of the promoter. The kinetics of the change in transcription show that the PT promoter can be shut off very rapidly by adding 50 mM MgSO4 to the medium, whereas resumption of transcription after removal of the modulative agents from the medium is slow. PMID- 2544568 TI - Symbiotic properties of rhizobia containing a flavonoid-independent hybrid nodD product. AB - A hybrid nodD gene consisting of 75% of the nodD1 gene of Rhizobium meliloti at the 5' end and 27% of the nodD gene of Rhizobium trifolii at the 3' end activates the six tested inducible nod promoters of Rhizobium leguminosarum, R. trifolii, or R. meliloti to maximal levels, even in the absence of flavonoids. In strains containing such a constitutive activating nodD gene, transcription of nod genes started at the same site as in flavonoid-induced strains containing a wild-type nodD gene. In contrast to heterologous wild-type nodD products, the constitutive activating nodD gene does not cause a limitation of the host range. Furthermore, R. leguminosarum, R. trifolii, and R. meliloti strains containing the constitutive activating nodD gene induce (pseudo) nodules on tropical leguminous plants. Comparison of the symbiotic properties of rhizobia containing the constitutive nodD hybrid gene with those of rhizobia containing various wild-type nodD genes indicates that the activation of the nodD product by flavonoids is of crucial importance during the process of infection thread formation and, surprisingly, also during nitrogen fixation. PMID- 2544569 TI - Methionine aminopeptidase gene of Escherichia coli is essential for cell growth. AB - We localized the methionine aminopeptidase (map) gene on the Escherichia coli chromosome next to the rpsB gene at min 4. Genetically modified strains with the chromosomal map gene under lac promoter control grew only in the presence of the lac operon inducer isopropyl-beta-thiogalactoside. Thus, methionine aminopeptidase is essential for cell growth. PMID- 2544570 TI - Mutagenesis of the Tra1 core region of RK2 by using Tn5: identification of plasmid-specific transfer genes. AB - The conjugation system of the IncP alpha plasmid RK2/RP4 is encoded by transfer regions designated Tra1, Tra2, and Tra3. The Tra1 core region, cloned on plasmid pDG4 delta 22, consists of the origin of transfer (oriT) and 2.6 kilobases of flanking DNA providing IncP alpha plasmid-specific functions that allow pDG4 delta 22 to be mobilized by the heterologous IncP beta plasmid R751. Tn5 insertions in pDG4 delta 22 define a minimal 2.2-kilobase region required for plasmid-specific transfer of oriT. The Tra1 core contains the traJ and traK genes as well as an 18-kilodalton open reading frame downstream of traJ. The traJ and traK genes were shown to be required for transfer by complementation of inserts within these genes. Genetic evidence for the role of the 18-kilodalton open reading frame in transfer was obtained, although this protein has not been detected in cell lysates. These studies indicate that at least three transfer proteins are involved in plasmid-specific interactions at oriT. PMID- 2544571 TI - An acute exercise-induced translocation of beta-adrenergic receptors in rat myocardium. AB - The effect of acute exercise (treadmill running) on rat myocardium beta adrenergic receptors (beta-AR) was studied. beta-AR was identified in purified sarcolemmal membrane fractions and light vesicle fractions. In control hearts, the number of beta-AR was 21.25 +/- 2.25 and 20.89 +/- 2.89 fmol/mg protein (mean +/- SE) in sarcolemmal membranes and light vesicles, respectively. Immediately after a single bout of dynamic exercise, about 35% of beta-AR was transferred from light vesicles to sarcolemmal membranes (p less than 0.05); concomitantly, isoproterenol-stimulated adenylate cyclase activity also significantly increased in sarcolemmal membranes (p less than 0.05). These results suggest that acute exercise provokes the translocation of beta-AR from a presumably intracellular site (light vesicles) to functional membrane fractions (sarcolemmal membranes) in rat myocardium. PMID- 2544572 TI - Characterization of specific high-affinity receptor for human lymphotoxin. AB - The specific cell surface receptors for lymphotoxin (LT) which are expressed on murine fibroblast L.P3 cells, a subline of L929 cells, were found to consist of a single class of specific high-affinity receptors with a dissociation constant (Kd) of 3.8 X 10(-10) M and a density of 5.8 X 10(3) sites/cell. Similarly, murine fibroblast L929 cells, human melanoma A375 cells and human cervical carcinoma HeLa-S3 cells had about 7.2 X 10(3), 3.5 X 10(3), and 6.6 X 10(3) sites/cell with Kd values of 1.4 X 10(-10), 0.5 X 10(-10), and 1.1 X 10(-10) M, respectively. Among the LT receptor-positive cell lines, there was no direct correlation between the level of specific LT binding and the sensitivity to the cytotoxic or cytostatic effect of LT. Cross-linking of 125I-LT to the cell surface receptors with disuccinimidyl suberate, followed by two-dimensional gel electrophoresis of the cell lysate, revealed two kinds of LT-LT receptor complexes with molecular weights of 70 and 97 kDa, and having the same pI value of 6.8. Cell-bound 125I-LT was internalized within 1 h and degraded intracellularly, and finally secreted into the medium within a few hours. Appropriate concentrations of LT and interferon gamma (IFN gamma) showed synergistic cytotoxicity toward murine fibroblast L.P3 cells and human monocytoma U937 cells, but these cytokines were only slightly cytotoxic individually. Preincubation of these cells with IFN gamma increased the total number of LT receptors without any significant change in the dissociation constant or in the molecular weight.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544573 TI - Concanavalin A receptor(s) possibly interacts with at least two kinds of GTP binding proteins in murine thymocytes. AB - A part of the GTP gamma S-binding activity in murine thymocyte membranes was found to have affinity to a concanavalin A (Con A)-Sepharose column. The material was identified as Gi (inhibitory GTP-binding protein) on the basis of the molecular weight and by islet activating protein-dependent ADP-ribosylation and anti-alpha i (alpha subunit of Gi) immunoblotting. However, when the membranes prepared from Con A-stimulated thymocytes were used, no GTP gamma S-binding activity was detected in the Con A-bound fraction, suggesting that Gi physically and specifically associated with Con A acceptors dissociates upon Con A stimulation. Furthermore, another GTP gamma S-binding protein (25 kDa), which is quite similar to a novel phosphoinositide-specific phospholipase C (PI-PLC) associated G protein in calf thymocytes (Wang, P., Toyoshima, S., & Osawa, T. (1988) J. Biochem. 103, 137-142), was detected among the Con A-Sepharose-bound proteins with the chemical cross-linking technique. When the 40 kDa and 25 kDa G proteins associated with Con A receptor(s) were isolated and their direct effects on the activity of partially purified PI-PLC as to phosphatidylinositol 4,5 bisphosphate hydrolysis were examined, the 25 kDa G protein was found to enhance the PI-PLC activity more effectively. On the other hand, pretreatment of cells with islet-activating protein completely abolished the inhibitory effect of Con A on the prostaglandin E1 and isoproterenol-induced increases of cellular cAMP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544574 TI - Dual cis-acting negative regulatory elements located upstream of the mouse DNA polymerase beta gene. AB - A 2,200 base pair (bp) fragment containing the 5'-flanking region of the mouse DNA polymerase beta gene was placed adjacent to and upstream of the chloramphenicol acetyl transferase (CAT)-coding region of the CAT vector. A transient expression assay of CAT activity in mouse NIH/3T3 cells transfected with this recombinant plasmid or a set of its 5'-deletion derivatives was carried out to identify a cis-acting regulatory element(s) for DNA polymerase beta gene expression. Depending on the extent of the deletion, CAT activity was dramatically increased, indicating the existence of a negative regulatory region which could be divided into two distinct domains: removal of the first domain (NRE-I), nucleotides -1860 to -1580 (+1 denotes the position of 5'-most proximal transcription initiation site), caused two to three-fold stimulation of CAT activity, and removal of the second domain (NRE-II), nucleotides -828 to -456, stimulated CAT expression another two to three-fold. When an 1,864-bp segment containing these negative regulatory elements (-2190 to -327) was inserted in the plasmid carrying the simian virus 40 early promoter and enhancer-directed CAT gene, it inhibited the CAT expression relatively independently of the orientation of insertion and the distance from the promoter-enhancer. We also mapped the promoter element of the DNA polymerase beta gene to within a 133-bp DNA fragment from nucleotide position -100 to +33. Either the NRE-I region or the NRE-II region alone can inhibit DNA polymerase beta gene promoter function. PMID- 2544575 TI - Amino acid sequence of the alpha and beta subunits of Methanosarcina barkeri ATPase deduced from cloned genes. Similarity to subunits of eukaryotic vacuolar and F0F1-ATPases. AB - The atpA and atpB genes coding for the alpha and beta subunits, respectively, of membrane ATPase were cloned from a methanogen Methanosarcina barkeri, and the amino acid sequences of the two subunits were deduced from the nucleotide sequences. The methanogenic alpha (578 amino acid residues) and beta (459 amino acid residues) subunits were highly homologous to the large and small subunits, respectively, of vacuolar H+-ATPases; 52% of the residues of the methanogenic alpha subunit were identical with those of the large subunit of vacuolar enzyme of carrot or Neurospora crassa, respectively, and 59, 60, and 59% of the residues of the methanogenic beta subunit were identical with those of the small subunits of N. crassa, Arabidopsis thaliana, and Sacharomyces cerevisiae, respectively. The methanogenic subunits were also highly homologous to the corresponding subunits of Sulfolobus acidocaldarius ATPase. The methanogenic alpha and beta subunits showed 22 and 24% identities with the beta and the alpha subunits of Escherichia coli F1, respectively. Furthermore, important amino acid residues identified genetically in the E. coli enzyme were conserved in the methanogenic enzyme. This sequence conservation suggests that vacuolar, F1, methanogenic, and S. acidocaldarius ATPases were derived from a common ancestral enzyme. PMID- 2544576 TI - The complete nucleotide sequence, gene organization, and genetic code of the mitochondrial genome of Paracentrotus lividus. AB - The 15,697-nucleotide sequence of Paracentrotus lividus mitochondrial DNA is reported. This genome codes for 2 rRNAs, 22 tRNAs, and 12 mRNAs which specify 13 subunits of the mitochondrial inner membrane respiratory complexes. The gene arrangement differs from that of other animal species. The two ribosomal genes 16 S and 12 S are separated by a stretch of about 3.3 kilobase pairs which contains the ND1 and ND2 genes and a cluster of 15 tRNA genes. The ND4L coding sequence is not contained in the ND4 mRNA but has its own mRNA which maps between the tRNA(Arg) and the Co II genes. The main noncoding region, located in the tRNA gene cluster, is only 132 nucleotides long, but contains sequences homologous to the mammalian displacement loop. Other short noncoding sequences are interspersed in the genome: they contain a conserved AT consensus which probably has a role in transcription or RNA processing. As regards the mitochondrial genetic code, the codons AGA and AGG specify serine and are recognized by a tRNA with a GCU anticodon, whereas AUA and AAA code for isoleucine and asparagine rather than for methionine and lysine. Except for ND4L which starts with AUC and ATPase 8 which starts with GUG, AUG is used as the initiation codon. In 11 out of 13 cases the genes terminate with the canonical stop codons UAA or UAG. These observations suggest that during invertebrate evolution each lineage developed its own mechanism of mitochondrial DNA replication and transcription and of RNA processing and translation. PMID- 2544577 TI - Beta 2-adrenergic agonist regulation of immune aggregate- and platelet-activating factor-stimulated hepatic metabolism. AB - Vasoconstriction and subsequent glycogenolysis stimulated by immune complex infusion into perfused rat livers was inhibited by prior infusion of isoproterenol. Similarly, isoproterenol inhibited the biosynthesis of bioactive lipid autacoids such as platelet-activating factor, prostaglandin E2, and thromboxane B2 which was stimulated by immune aggregates. The adrenergic receptor specificity of these effects was determined through the use of specific adrenergic subtype-specific agonists and antagonists to be mediated by beta 2 adrenergic receptors. Indirect evidence for the differential expression of hepatic sinusoidal and parenchymal beta-adrenergic receptors in the male rat during ontogeny suggested that inhibition of immune aggregate-stimulated autacoid biosynthesis, vasoconstriction, and glycogenolysis by isoproterenol occurs at a sinusoidal locus, most likely Kupffer cells. In contrast with the ability of beta 2-adrenergic agonists to inhibit immune aggregate- and platelet-activating factor stimulated hepatic metabolism, dibutyryl cyclic AMP did not mimic these sinusoidal beta 2-adrenergic effects, despite stimulating hepatic parenchymal cell glycogenolysis as effectively as isoproterenol. These observations suggest a role for cyclic AMP-independent mechanisms in the regulation of heterologous stimulus-response coupling by hepatic sinusoidal beta 2-adrenergic receptors. PMID- 2544579 TI - Structure and function of human hemoglobin covalently labeled with periodate oxidized adenosine triphosphate. AB - Periodate-oxidized adenosine triphosphate (o-ATP), a ribose ring-opened dialdehyde derivative of ATP, reacts specifically with human deoxyhemoglobin to give a single major covalently modified product after reduction with sodium borohydride. This product, designated di-ATP Hb, was isolated using ion-exchange chromatography and shown to have incorporated two molecules of o-ATP/tetramer. Peptide mapping and x-ray crystallography at 2.8-A resolution indicate that a covalent adduct is formed between the ligand and residues Lys-82 EF6 of each beta chain in the organic phosphate-binding site of the molecule. di-ATP Hb exhibits a significantly decreased oxygen affinity (P50 = 20.8 mm Hg versus 5.8 mm Hg control; 50 mM 2-[bis(2-hydroxyethyl)amino]-2-(hydroxymethyl)-propane-1,3-diol, pH 7.4, 0.1 M C, 20 degrees C). The subunit cooper-activity of di-ATP Hb is also reduced (nmax = 1.9 versus 2.7 control). PMID- 2544578 TI - Coding nucleotide, 5' regulatory, and deduced amino acid sequences of P-450BM-3, a single peptide cytochrome P-450:NADPH-P-450 reductase from Bacillus megaterium. AB - Cytochrome P-450BM-3 (P-450BM-3) from Bacillus megaterium incorporates both a P 450 and an NADPH:P-450 reductase in proteolytically separable domains of a single, 119-kDa polypeptide and functions as a fatty acid monooxygenase independently of any other protein. A 5-kilobase DNA fragment which contains the gene encoding P-450BM-3 was sequenced. A single continuous open reading frame starting at nucleotide 1541 of the 5-kilobase fragment correctly predicted the previously determined NH2-terminal protein sequences of the trypsin-generated P 450 and reductase domains and, in toto, predicted a mature polypeptide of 1,048 amino acid residues with Mr = 117,641. The trypsin site was found at arginine residue 471. The P-450 domain is most similar (about 25%) to the fatty acid omega hydroxylases of P-450 family IV, while the reductase domain exhibits some 33% sequence similarity with the NADPH:P-450 reductases of mammalian liver. Both the P-450 and reductase domains of P-450BM-3 define new gene families but contain highly conserved segments which display as much as 50% sequence similarity with P 450s and reductases of eukaryotic origin. The mRNA for P-450BM-3 was found by S1 mapping to be 3,339 +/- 10 nucleotides in length. In the accompanying paper, two regions in the 1.5 kilobases 5' to the P-450BM-3 coding region have been implicated in the regulation of P-450BM-3 gene expression. PMID- 2544580 TI - Characterization of semiquinone free radicals formed from stilbene catechol estrogens. An ESR spin stabilization and spin trapping study. AB - Electron spin resonance spectroscopy has been used to detect, characterize, and to infer structures of o-semiquinones derived from stilbene catechol estrogens. Radicals were generated enzymatically using tyrosinase and were detected as their Mg2+ complexes. It is suggested that initial hydroxylation of stilbene estrogen gives a catechol estrogen in situ; subsequent two-electron oxidation of the catechol to the quinone, followed by reverse disproportionation, leads to the formation of radicals. Consistent with this mechanism, o-phenylenediamine, a quinone trapping agent, inhibits formation of o-semiquinones. A competing mechanism of radical production involves autoxidation of the catechol. Hydroxyl radicals are shown to be produced in this system via a mechanism involving reduction of iron and copper complexes by stilbene catechols. Possible differences in the reactivity of stilbene ortho- and para-semiquinones are discussed. PMID- 2544581 TI - Structure of the gene for the arylphorin-type storage protein SP 2 of Bombyx mori. AB - Two forms of storage proteins termed "SP 1" and "SP 2", respectively, accumulate in a stage-dependent fashion in the larval hemolymph of the silkworm Bombyx mori. We have cloned and analyzed the genomic sequence encoding SP 2, an arylphorin type storage protein. The SP 2 structural gene contains five exons spanning over 5 kilobases of chromosomal DNA. The transcription initiation site of the SP 2 gene was identified at the nucleotide level and a typical TATA box was located 30 base pairs upstream of the transcription initiation site. The 5'-flanking region of the gene contains a sequence homologous to the SV 40 enhancer "core" structure. A region homologous to the Drosophila enhancer sequence predicted for the fat body specificity of gene expression exists in the 5'-upstream region. Developmental profiles of the SP 2 mRNA and its precursor in the fat body indicate that the biosynthesis of SP 2 is developmentally regulated at the level of transcription. The exon/intron composition of the SP 2 gene is remarkably similar to that of the SP 1 gene. Moreover, the deduced primary structures of SP 1 and SP 2 exhibit nearly 30% homology, implying that two storage protein genes of the B. mori silkworm might have evolved from a common ancestor. PMID- 2544583 TI - The central part of parathyroid hormone stimulates thymidine incorporation of chondrocytes. AB - The stimulation of DNA synthesis in primary cell cultures of chicken chondrocytes by parathyroid hormone was studied by assaying [3H]thymidine incorporation into DNA. Optimal assay conditions were determined by varying cell age, plating density, and incubation time. Under these conditions DNA synthesis was significantly stimulated by parathyroid hormone (PTH) and some of its fragments: cells treated with human (h)PTH(1-84), bovine (b)PTH(1-34) and [Nle8,18,Tyr34]bPTH(3-34)amide and hPTH(13-34) displayed 2.6-fold enhanced [3H]thymidine incorporation in a dose-dependent manner. The fragment hPTH(28-48) led to a similar stimulation, whereas [Tyr43]hPTH(43-68) and [Tyr52,Asp76]hPTH(52 84) had no effect. Using a series of synthetic hPTH peptides covering the central region of the hormone molecule (residues 25-47), we could delimitate further this putative mitogenic functional domain to a core region between amino acid residues 30 and 34. The effect of PTH on [3H]thymidine incorporation could not be mimicked by forskolin, indicating that the corresponding signal is not mediated by cAMP. It is, however, inhibited by EGTA and cannot be provoked in the absence of calcium ions in the medium. Therefore, the results presented indicate a hitherto unidentified functional domain of PTH in the central part of the molecule which exerts its mitogenic effect on chondrocytes in a cAMP-independent manner but seems to involve calcium ions for signal transduction. PMID- 2544582 TI - Antigonadotropic effects of the bovine ovarian gonadotropin-releasing hormone binding inhibitor/histone H2A in rat luteal and granulosal cells. AB - A gonadotropin-releasing hormone (GnRH)-binding inhibitor (GnRH-BI) was purified from bovine ovaries and identified as histone H2A. In the present studies, the biological effects of partially purified and purified ovarian GnRH-BI, as well as calf thymus histone H2A, were examined in rat ovarian cells. Since GnRH has direct antigonadotropic actions on these cells, the effects on luteinizing hormone-stimulated cAMP accumulation in luteal cells and follicle stimulating hormone-induced cAMP and progesterone production in granulosal cells were evaluated. Antigonadotropic activity in both luteal and granulosal cells coeluted directly with GnRH-BI activity during purification from bovine ovaries, and the antigonadotropic effects were dose dependent and reversible. In contrast to GnRH, GnRH-BI maximally inhibited gonadotropin responses and the effects of GnRH-BI were not blocked by a GnRH antagonist. The purified ovarian GnRH-BI and calf thymus histone H2A had identical antigonadotropic properties, and the half maximal concentrations for inhibiting the gonadotropin responses of granulosal and luteal cells was 2 and 5 microM, respectively. In conclusion, the ovarian GnRH-binding inhibitor, identified as histone H2A, not only inhibits the high affinity binding of GnRH to rat ovarian membranes but also evokes GnRH-like antigonadotropic responses in rat ovarian cells that do not appear to be mediated by binding to GnRH receptors. PMID- 2544584 TI - Unusual composition of peptidoglycan in Bordetella pertussis. AB - The composition of the peptidoglycan of Bordetella pertussis and the nature of its turnover products was determined by a new combination of analytical techniques: high performance liquid chromatography of an enzymatic peptidoglycan hydrolysate and fast atom bombardment mass spectrometry and fast atom bombardment collision-activated dissociation tandem mass spectrometry. Sixteen major components of the peptidoglycan were purified, and assignment of complete or partial chemical structures was achieved for nine and seven species, respectively. At this level of resolution, a previously unrecognized heterogeneity of monomeric (five new species; nine total) and dimeric species (five new species; five total) was detected. No species containing diaminopimelyl diaminopimelic acid cross-links or lysyl-arginine substitutions were found. Previous estimates of total cross-linkage and average chain length were revised downward to 32% and 21 disaccharide residues, respectively. Detection of a chemically novel species, a disaccharide octapeptide monomer, in both the peptidoglycan hydrolysate and culture supernatant fluid, suggests that an N acetyl-muramyl-L-alanine amidase acts on the intact peptidoglycan of Bordetella and participates in cell wall turnover. Five peptidoglycan turnover products were identified in the supernatant fluid of late logarithmic phase cultures, including the 1,6-anhydro monomeric species known as tracheal cytotoxin. Peptidoglycan turnover was detected at a low rate of approximately 10%/generation, a value sufficient to account for the generation of all tracheal cytotoxin found in culture supernatant fluids. PMID- 2544585 TI - Macromolecular specificity determinants on thrombin for fibrinogen and thrombomodulin. AB - The endothelial cell surface membrane protein thrombomodulin binds thrombin with high affinity and acts as both a cofactor for protein C activation and an inhibitor of fibrinogen hydrolysis. We have previously shown that bovine thrombomodulin is a competitive inhibitor of fibrinogen binding to thrombin but has no effect on thrombin activity toward tripeptide substrates or antithrombin III. Hence, thrombomodulin and fibrinogen may share macromolecular specificity sites on thrombin which are distinct from the active site. In this investigation, we have studied the interaction of thrombin-thrombomodulin with fibrinogen and various thrombin derivatives. We show that fibrinogen is a competitive inhibitor of thrombomodulin binding to thrombin, with a Kis = 10 microM. Thrombin derivatives (bovine (pyridoxal phosphate)4-thrombin and human thrombin Quick I), which bind fibrinogen with much reduced affinity, are shown to also interact with thrombomodulin with greatly reduced affinity. These results are consistent with the hypothesis that thrombomodulin and fibrinogen share macromolecular specificity sites on thrombin. PMID- 2544586 TI - Identification and characterization of a new gene (CBP3) required for the expression of yeast coenzyme QH2-cytochrome c reductase. AB - Respiratory defective mutants of Saccharomyces cerevisiae assigned to complementation group G28 display a deficiency, in the respiratory chain complex coenzyme QH2-cytochrome c reductase. The mutants define a new nuclear gene, designated CBP3, required for the assembly of the complex. Mutations in CBP3 are expressed in the absence of spectrally and immunologically detectable cytochrome b, a catalytic subunit of coenzyme QH2-cytochrome c reductase. The mutational block responsible for the cytochrome b deficiency has been ascribed to a post translational step based on the observation that cbp3 mutants have wild type concentrations of cytochrome b mRNA and are capable of synthesizing the apoprotein. Western analysis has revealed that cbp3 mutants have reduced levels of a subset of subunit polypeptides of the coenzyme QH2-cytochrome c reductase complex that include apocytochrome b, the iron-sulfur protein, core 4 (14-kDa subunit), and core 5 (11-kDa subunit). A similar phenotype has previously been reported in strains that fail to assemble the complex as a result of mutations in the noncatalytic core subunits. The CBP3 gene has been cloned by transformation of a mutant from complementation group G28 with a yeast genomic library. The gene is 1005 nucleotides long and codes for a primary translation product of 39 kDa. A transcript of a size commensurate with the length of the CBP3 reading frame is detected in total and poly(A+)-enriched RNA. The amino-terminal region of the CBP3 product is basic and probably corresponds to a cleavable mitochondrial targeting signal. An antibody obtained against a trpE/CBP3 fusion protein detects a protein of 40 kDa in wild type yeast mitochondria. This protein is absent in a mutant construct containing a partially deleted copy of the gene. The CBP3 protein is a membrane constituent, although attempts to demonstrate its physical association with the other subunits of coenzyme QH2-cytochrome c reductase have been unsuccessful. PMID- 2544587 TI - Characterization of the promoter region of the human thrombospondin gene. DNA sequences within the first intron increase transcription. AB - Thrombospondin (TSP) is an extracellular matrix glycoprotein whose synthesis and secretion by mesenchymal cells is regulated at the level of gene transcription by platelet-derived growth factor. To examine the transcriptional regulation of the TSP gene at the molecular level, a genomic clone containing the human TSP promoter and flanking sequence was isolated and characterized. A 3.8-kilobase pair (kb) DNA fragment containing the first three exons, the first two introns, and 2.2 kb of 5'-flanking region was sequenced, and the site of transcription initiation was determined by both primer extension and S1 nuclease mapping. Consensus sequences for several potential regulatory elements were found in the 5'-flanking sequence, including a TATA box consensus sequence, TTTAAAA, located 24 base pairs upstream from the transcription start site. A chimeric gene was constructed containing the first intron, the first exon, and 2.0 kb of 5' flanking sequence of the TSP gene fused to the promoterless gene for chloramphenicol acetyltransferase. When transfected into COS-1 or NIH3T3 cells this gene construct was transcribed, indicating the presence of a functional promoter in the TSP sequence. Transient transfection studies using deletion mutants of this TSP-chloramphenicol acetyltransferase construct were performed to locate cis-acting regulatory sequences. The deletion of flanking sequence 5' to position -234 had little or no effect on transcriptional activity, whereas deletion of 5'-flanking sequence extending further in the 3' direction resulted in the gradual loss of transcriptional activity. The removal of the first intron resulted in a 4-fold decrease in transcript levels, indicating the presence of a cis-acting positive element(s) in the first intron of the human TSP gene. This element(s) was further localized to the region between position +576 and position +727. PMID- 2544588 TI - Carboxyl-terminal modification of a gastrin releasing peptide derivative generates potent antagonists. AB - Gastrin releasing peptide (GRP) is a 27-residue peptide hormone which is analogous to the amphibian peptide bombesin. GRP serves a variety of physiological functions and has been implicated as an autocrine factor in the growth regulation of small cell lung cancer cells. We have developed a series of potent GRP antagonists by modification of the COOH terminus of N-acetyl-GRP-20 27. The most potent member of this series, N-acetyl-GRP-20-26-OCH2CH3, exhibits an IC50 of 4 nM in a competitive binding inhibition assay. This compound blocks GRP-stimulated mitogenesis in Swiss 3T3 mouse fibroblasts, inhibits GRP-dependent release of gastrin in vitro, and blocks GRP-induced elevation of [Ca2+]i in H345 small cell lung cancer cells. These results demonstrate that while residues 20-27 of GRP influence binding of the parent peptide to its receptor, the COOH-terminal amino acid is primarily responsible for triggering the subsequent biological response. PMID- 2544589 TI - Heparin binding domain of human antithrombin III inferred from the sequential reduction of its three disulfide linkages. An efficient method for structural analysis of partially reduced proteins. AB - Human antithrombin III (AT-III) was partially reduced under mild conditions in the absence or presence of low molecular weight heparin. Quantitation of reduced disulfide bonds was facilitated by the application of a water-soluble color reagent, 4-N,N-dimethylaminoazobenzene-4'-iodoacetamido-2'-sulfonic acid (S DABIA). The study shows that the three disulfide linkages of AT-III can be sequentially reduced, with Cys8-Cys128 being the most sensitive, followed by Cys21-Cys95, while Cys247-Cys430 is the most resistant to the mild reduction conditions. The rate of reduction of Cys8-Cys128 and Cys21-Cys95 was significantly decreased in the presence of heparin. The reduction of Cys8-Cys128 was also found to correlate quantitatively with the loss of heparin-accelerated antithrombin activity, heparin binding affinity, and heparin-induced fluorescence enhancement. These results suggest that Cys8-Cys128 is required for the integrity of the heparin binding domain of AT-III and support previous findings that lysyl residues surrounding Cys128 (Lys107, Lys114, Lys125, and Lys136) constitute an important part of the heparin binding site in AT-III. PMID- 2544590 TI - Mammalian topoisomerase II activity is modulated by the DNA minor groove binder distamycin in simian virus 40 DNA. AB - DNA topoisomerases II are nuclear enzymes that have been identified recently as targets for some of the most active anticancer drugs. Antitumor topoisomerase II inhibitors such as teniposide (VM-26) produce enzyme-induced DNA cleavage and inhibition of enzyme activity. By adding to such reactions distamycin, a compound whose effects on DNA have been extensively characterized, we investigated the effects of drug binding upon topoisomerase II-mediated DNA cleavage induced by VM 26. We have found a correspondence between distamycin binding (determined by footprinting analysis) and topoisomerase II-mediated cleavage of SV40 DNA (determined by sequencing gel analysis). Distamycin binding potentiated the cleavage of specific sites in the near proximity of distamycin-binding sites (within at least 25 base pairs), which indicates that DNA secondary structure is involved in topoisomerase II-DNA interactions. That distamycin potentiated cleavage only at sites that were recognized in the absence of distamycin and suppressed cleavage directly at distamycin-binding sites indicates that topoisomerase II recognizes DNA on the basis of primary sequence. In addition, distamycin stimulated topoisomerase II-mediated DNA relaxation and antagonized the inhibitory effect of VM-26. These results show that the DNA sequence-specific binding of distamycin produces local and propagated effects in the DNA which markedly affect topoisomerase II activity. PMID- 2544591 TI - Acid secretagogue-induced stimulation of gastric parietal cell gene expression. AB - Carbamoylcholine (carbachol), histamine, and gastrin are three principal stimulants of gastric acid secretion. To explore the mechanisms by which these agents exert their actions in parietal cells, we examined their effects on the gene expression of the enzymes responsible for H+ generation. Each secretagogue induced rapid and coordinate increases in steady-state levels of mRNAs encoding carbonic anhydrase II and H+,K+-ATPase in isolated canine gastric parietal cells. Furthermore, pronounced increases, with different kinetics, in expression of beta actin mRNA were observed. With increasing time after cell isolation, carbonic anhydrase II and H+,K+-ATPase, but not beta-actin, mRNA levels were attenuated, suggesting that parietal cell-specific genes may be dependent upon maintenance of parietal cell contacts within intact mucosal tissue. Pretreatment of the cells with competitive inhibitors of each secretagogue blocked the increases. Our results indicate that acid secretagogue-specific receptor activation in parietal cells triggers coordinate gene expression of the two enzymes involved in H+ ion generation and that beta-actin may be an important regulator of acid secretion. PMID- 2544592 TI - Inhibition of the high affinity Fc receptor (Fc gamma RI) on human monocytes by porphyrin photosensitization is highly specific and mediated by the generation of superoxide radicals. AB - p72 high affinity receptors (Fc gamma RI) for the Fc portion of IgG molecules on human peripheral blood monocytes mediate a variety of beneficial functions, but also have deleterious effects in certain clinical situations. In the present study, the photosensitizing porphyrins hematoporphyrin derivative and dihematoporphyrin ether (DHE), which are known to preferentially affect the cell membrane, were found to significantly inhibit binding of mouse IgG2a antibodies to the ligand binding site of Fc gamma RI on human peripheral blood monocytes and the U937 human monocytic cell line. Fc gamma RI receptors could be identified with a monoclonal antibody which recognizes an epitope distinct from the ligand binding site, indicating that photosensitization induced a structural alteration rather than loss of the receptor molecule from the cell surface. The effect of DHE and light appeared to be highly specific, since binding of monoclonal antibodies to other surface structures was not decreased. DHE plus light-induced modulation of Fc gamma RI was found to be mediated by superoxide anions, since addition of a mimic of superoxide dismutase restored both binding of mouse IgG2a to Fc gamma RI as well as human monocyte accessory cell function. These studies identify porphyrin photosensitization as a unique mechanism by which to selectively down-regulate Fc gamma RI-mediated functions. PMID- 2544593 TI - Regulation of expression of the gene encoding the major surfactant protein (SP-A) in human fetal lung in vitro. Disparate effects of glucocorticoids on transcription and on mRNA stability. AB - We previously observed that dexamethasone had a biphasic effect on the levels of mRNA encoding the major surfactant protein (SP-A) in human fetal lung in vitro; at concentrations of 10(-10) and 10(-9) M, dexamethasone caused an increase in the levels of SP-A mRNA, whereas at concentrations of greater than 10(-8) M, the steroid had a pronounced inhibitory effect on SP-A mRNA levels. It was also found that dexamethasone antagonized the stimulatory effect of dibutyryl cyclic AMP (Bt2cAMP) on SP-A mRNA levels in human fetal lung in vitro. It was our objective, in the present study, to characterize further the effects of dexamethasone and Bt2cAMP on SP-A mRNA levels in human fetal lung tissue and to determine whether such effects are associated with comparable changes in the transcriptional activity of the SP-A gene. We found that the action of dexamethasone (10(-7) M) to reduce the levels of SP-A mRNA in control and Bt2cAMP-treated fetal lung explants was evident within 2 h of its addition to the culture medium; SP-A mRNA was reduced to barely detectable levels in control and in Bt2cAMP-treated tissues after 24 h of dexamethasone treatment. The action of dexamethasone to reduce SP-A mRNA levels was not prevented by co-incubation with either actinomycin D or cycloheximide. In contrast to its dose-related biphasic effects on the levels of SP-A mRNA, we found that dexamethasone caused a dose-dependent stimulation of SP A gene transcription. Bt2cAMP also increased the transcriptional activity of the SP-A gene in the human fetal lung in vitro. In fetal lung explants incubated in the presence of dexamethasone plus Bt2cAMP, a synergistic induction of SP-A gene transcription was observed at concentrations of dexamethasone of 10(-9)-10(-7) M. Our findings are indicative that the stimulatory effects of dexamethasone (10( 10)-10(-9) M) on SP-A mRNA levels are reflective of a stimulatory effect of the steroid on SP-A gene transcription, whereas the inhibitory effects of dexamethasone (10(-7) M) on SP-A mRNA levels are the result of a dominant effect of the steroid in elevated concentrations to reduce SP-A mRNA stability. PMID- 2544594 TI - The cation-dependent mannose 6-phosphate receptor. Structural requirements for mannose 6-phosphate binding and oligomerization. AB - The structural requirements for oligomerization and the generation of a functional mannose 6-phosphate (Man-6-P) binding site of the cation-dependent mannose 6-phosphate receptor (CD-MPR) were analyzed. Chemical cross-linking studies on affinity-purified CD-MPR and on solubilized membranes containing the receptor indicate that the CD-MPR exists as a homodimer. To determine whether dimer formation is necessary for the generation of a Man-6-P binding site, a cDNA coding for a truncated receptor consisting of only the signal sequence and the extracytoplasmic domain was constructed and expressed in Xenopus laevis oocytes. The expressed protein was completely soluble, monomeric in structure, and capable of binding phosphomannosyl residues. Like the dimeric native receptor, the truncated receptor can release its ligand at low pH. Ligand blot analysis using bovine testes beta-galactosidase showed that the monomeric form of the CD-MPR from bovine liver and testes is capable of binding Man-6-P. These results indicate that the extracytoplasmic domain of the receptor contains all the information necessary for ligand binding as well as for acid-dependent ligand dissociation and that oligomerization is not required for the formation of a functional Man-6-P binding site. Several different mutant CD-MPRs were generated and expressed in X. laevis oocytes to determine what region of the receptor is involved in oligomerization. Chemical cross-linking analyses of these mutant proteins indicate that the transmembrane domain is important for establishing the quaternary structure of the CD-MPR. PMID- 2544595 TI - Phospholamban phosphorylation in intact ventricles. Phosphorylation of serine 16 and threonine 17 in response to beta-adrenergic stimulation. AB - Phospholamban is the major membrane protein of the heart phosphorylated in response to beta-adrenergic stimulation. In cell-free systems, cAMP-dependent protein kinase catalyzes exclusive phosphorylation of serine 16 of phospholamban, whereas Ca2+/calmodulin-dependent protein kinase gives exclusive phosphorylation of threonine 17 (Simmerman, H. K. B., Collins, J. H., Theibert, J. L., Wegener, A. D., and Jones, L. R. (1986) J. Biol. Chem. 261, 13333-13341). In this work we have localized the sites of phospholamban phosphorylation in intact ventricles treated with the beta-adrenergic agonist isoproterenol. Isolation of phosphorylated phospholamban from 32P-perfused guinea pig ventricles, followed by partial acid hydrolysis and phosphoamino acid analysis, revealed phosphorylation of both serine and threonine residues. At steady state after isoproterenol exposure, phospholamban contained approximately equimolar amounts of these two phosphoamino acids. Two major tryptic phosphopeptides containing greater than 90% of the incorporated radioactivity were obtained from phospholamban labeled in intact ventricles. The amino acid sequences of these two tryptic peptides corresponded exactly to residues 14-25 and 15-25 of canine cardiac phospholamban, thus localizing the sites of in situ phosphorylation to serine 16 and threonine 17. Phosphorylation of phospholamban at two sites in heart perfused with isoproterenol was supported by detection of 11 distinct mobility forms of the pentameric protein by use of the Western blotting method, consistent with each phospholamban monomer containing two phosphorylation sites, and with each pentamer containing from 0 to 10 incorporated phosphates. Our results localize the sites of in situ phospholamban phosphorylation to serine 16 and threonine 17 and, furthermore, are consistent with the phosphorylations of these 2 residues being catalyzed by cAMP- and Ca2+/calmodulin-dependent protein kinases, respectively. PMID- 2544596 TI - A monoclonal antibody against the rod outer segment guanyl nucleotide-binding protein, transducin, blocks the stimulatory and inhibitory G proteins of adenylate cyclase. AB - GTP-binding proteins have been implicated as transducers of a variety of biological signaling processes. These proteins share considerable structural as well as functional homology. Due to these similarities, it was thought that a monoclonal antibody that inhibits the light activation of the rod outer segment GTP-binding protein, tranducin (Gt), might exert some functional effect upon the G proteins that regulate the adenylate cyclase system. Antibody 4A, raised against the alpha subunit of Gt, cross-reacted (by hybridization on nitrocellulose) with purified alpha subunits of other G proteins (Gi and Gs, regulatory guanyl nucleotide-binding proteins that mediate inhibition and stimulation of adenylate cyclase, respectively) as long as they were not denatured. This antibody, which interferes with rod outer segment cGMP phosphodiesterase activation by blocking interaction between rhodopsin and Gt, also interfered with actions of both the stimulatory and inhibitory G proteins of adenylate cyclase from rat cerebral cortex membranes. Effects of monoclonal antibody (mAb) 4A were dose-dependent and not reversed by washing. mAb 4A also blocked the Gi-mediated inhibition of adenylate cyclase in the cyc- variant of S49 lymphoma and in doing so raised the level of adenylate cyclase activity in both the cyc- variant and the S49 wild type. There was no effect of mAb 4A on adenylate cyclase activity of the resolved catalytic subunit. These results suggest that the well known sequence homologies among the G proteins involved in cellular signal transduction may extend to the sites that interact with other members of signal-transducing cascades (receptors and effector molecules). Therefore, antibody 4A may serve as a useful tool to probe the similarities and differences among the various systems. PMID- 2544597 TI - Identification and properties of very high affinity brain membrane-binding sites for a neurotoxic phospholipase from the taipan venom. AB - Four new monochain phospholipases were purified from the Oxyuranus scutellatus (taipan) venom. Three of them were highly toxic when injected into mice brain. One of these neurotoxic phospholipases, OS2, was iodinated and used in binding experiments to demonstrate the presence of two families of specific binding sites in rat brain synaptic membranes. The affinities were exceptionally high, Kd1 = 1.5 +/- 0.5 pM and Kd2 = 45 +/- 10 pM, and the maximal binding capacities were Bmax 1 = 1 +/- 0.4 and Bmax 2 = 3 +/- 0.5 pmol/mg of protein. Both binding sites were sensitive to proteolysis and demonstrated to be located on proteins of Mr 85,000-88,000 and 36,000-51,000 by cross-linking and photoaffinity labeling techniques. The binding of 125I-OS2 to synaptic membranes was dependent on Ca2+ ions and enhanced by Zn2+ ions which inhibit phospholipase activity. Competition experiments have shown that, except for beta-bungarotoxin, a number of known toxic snake or bee phospholipases have very high affinities for the newly identified binding sites. A good correlation (r = 0.80) was observed between toxicity and affinity but not between phospholipase activity and affinity. PMID- 2544598 TI - Effect of phosphorothioate homo-oligodeoxynucleotides on herpes simplex virus type 2-induced DNA polymerase. AB - Effects of phosphorothioate oligodeoxynucleotides of different chain length and base composition on herpes simplex virus (HSV) type 2 (strain 333)-induced DNA polymerase have been examined in vitro. The anti-HSV-2 DNA polymerase activity was related to the base composition of the analogs, with the order of potency: deoxycytidine greater than thymidine greater than deoxyadenosine, for compounds with equal chain length. The potency was also related to oligomer chain length, since it was observed that the longer the chain length, the more potent the inhibition exerted. Among all the compounds tested, the phosphorothioate oligodeoxycytidine 28-mer (S-(dC)28) was the most potent inhibitor of HSV-2 induced DNA polymerase. This inhibition was competitive with an activated DNA template with a Ki value of 7 nM. It was also a competitive inhibitor of the DNA polymerase-associated exonuclease activity with a Ki value of 5 nM. In contrast, this compound showed less inhibition of human DNA polymerase alpha, beta, and gamma, as well as HSV-1 (strain KOS) and Epstein-Barr virus-induced DNA polymerase. The possibility that S-oligomers can serve as primers for DNA elongation was also investigated. Poly(dG).S-(dC)28 and poly(dA).S-(T)28 are poor substrates for DNA elongation catalyzed by HSV-2 DNA polymerase. In summary, phosphorothioate oligonucleotides could be anti-template inhibitors of HSV DNA polymerase. This information may lead to the development of a new class of selective anti-HSV agents. PMID- 2544599 TI - Heat shock of rabbit synovial fibroblasts increases expression of mRNAs for two metalloproteinases, collagenase and stromelysin. AB - Two metalloproteinases, collagenase and stromelysin, are produced in large quantities by synovial fibroblasts in individuals with rheumatoid arthritis. These enzymes play a major role in the extensive destruction of connective tissue seen in this disease. In this study, we show that heat shock of monolayer cultures of rabbit synovial fibroblasts increases expression of mRNA for heat shock protein 70 (HSP-70), and for collagenase and stromelysin. We found that after heat shock for 1 h at 45 degrees C, the mRNA expression for HSP-70 peaks at 1 h and returns to control levels by 3 h. Collagenase and stromelysin mRNA expression is coordinate, reaching peak levels at 3 h and returning to control levels by 10 h. The increase in mRNA is paralleled by an increase in the corresponding protein in the culture medium. 3 h of heat shock at a lower temperature (42 degrees C) is also effective in inducing collagenase and stromelysin mRNAs. Concomitant treatment with phorbol myristate acetate (PMA; 10( 8) or 10(-9) M) and heat shock is not additive or synergistic. In addition, all trans-retinoic acid, added just before heat shock, prevents the increase in mRNAs for collagenase and stromelysin. Our data suggest that heat shock may be an additional mechanism whereby collagenase and stromelysin are increased during rheumatoid arthritis and perhaps in other chronic inflammatory stress conditions. PMID- 2544600 TI - Lamin A, lamin B, and lamin B receptor analogues in yeast. AB - Previous studies have shown that turkey erythrocyte lamin B is anchored to the nuclear envelope via a 58-kD integral membrane protein termed p58 or lamin B receptor (Worman H. J., J. Yuan, G. Blobel, and S. D. Georgatos. 1988. Proc. Natl. Acad. Sci. USA. 85:8531-8534). We now identify a p58 analogue in the yeast Saccharomyces cerevisiae. Turkey erythrocyte lamin B binds to yeast urea extracted nuclear envelopes with high affinity, associating predominantly with a 58-kD polypeptide. This yeast polypeptide is recognized by polyclonal antibodies against turkey p58, partitions entirely with the nuclear fraction, remains membrane bound after urea extraction of the nuclear envelopes, and is structurally similar to turkey p58 by peptide mapping criteria. Using polyclonal antibodies against turkey erythrocyte lamins A and B, we also identify two yeast lamin forms. The yeast lamin B analogue has a molecular mass of 66 kD and is structurally related to erythrocyte lamin B. Moreover, the yeast lamin B analogue partitions exclusively with the nuclear envelope fraction, is quantitatively removed from the envelopes by urea extraction, and binds to turkey lamin A and vimentin. As many higher eukaryotic lamin B forms, the yeast analogue is chemically heterogeneous comprising two serologically related species with different charge characteristics. Antibodies against turkey lamin A detect a 74 kD yeast protein, slightly larger than the turkey lamin A. It is more abundant than the yeast lamin B analogue and partitions between a soluble cytoplasmic fraction and a nuclear envelope fraction. The yeast lamin A analogue can be extracted from the nuclear envelope by urea, shows structural similarity to turkey and rat lamin A, and binds to isolated turkey lamin B. These data indicate that analogues of typical nuclear lamina components (lamins A and B, as well as lamin B receptor) are present in yeast and behave as their vertebrate counterparts. PMID- 2544601 TI - Secretion in yeast: preprotein binding to a membrane receptor and ATP-dependent translocation are sequential and separable events in vitro. AB - We have used a cytosol-free assay in which efficient translocation and signal peptide cleavage is achieved when the affinity-purified precursor of OmpA (proOmpA) is diluted out of 8 M urea into a suspension of yeast rough microsomes. This aspect of protein targeting and transport occurs in two discernible steps: (a) in the absence of ATP and cytosolic factors, the precursor binds to the membranes but is not translocated; (b) addition of ATP results in the translocation of the bound precursor and its processing to the mature form. The binding to microsomes of radiolabeled proOmpA is saturable and inhibited by the addition of unlabeled proOmpA but not by mature OmpA or other proteins. The binding of radiolabeled prepro-alpha-factor is also effectively competed by other preproteins, but not by mature ones. Scatchard analysis showed the Kd of proOmpA to be 7.5 X 10(-9) M. Binding is most likely protein mediated as treatment of the microsomes with the protease papain was found to be inhibitory. These results represent the first functional characterization of secretory protein precursor binding to membranes. Alkylation of the microsomes with NEM, washing the membranes with urea or using membranes from the (translocation) mutant ptll at the nonpermissive temperature, did not affect binding, but did eliminate the subsequent ATP-dependent translocation. The ability to subdivide translocation into individual reactions provides a more precise means of determining the membrane components involved in this process. PMID- 2544604 TI - Evidence that aggregation of mouse sperm receptors by ZP3 triggers the acrosome reaction. AB - In the mouse, considerable evidence indicates that initial sperm binding to the zona pellucida (ZP) is mediated by ZP3. In addition, this same glycoprotein is also responsible for inducing the acrosome reaction (AR). Whereas the O-linked oligosaccharides of ZP3 appear to mediate sperm-ZP binding, the portion of ZP3 bearing AR activity has not been defined. To try to understand the bifunctional role of ZP3 (binding and AR inducing activities), we have examined the hypothesis that ZP3 aggregates sperm receptor molecules. By analogy with findings in a variety of other extracellular signal transducing systems, including receptors for growth factors and insulin, this aggregation event could initiate the cascade resulting in the AR. To test this hypothesis, we have generated monospecific polyclonal antibodies against ZP2 and against ZP3, and examined the effects of these probes on capacitated sperm incubated in the absence or presence of various ZP protein preparations. For some experiments, we have used proteolytic fragments of ZP3, a preparation known to retain specific binding, but not AR-inducing, activity. We show here that capacitated mouse sperm, incubated with ZP glycopeptides, displayed ARs when incubated subsequently with anti-ZP3 IgG; ARs did not occur when parallel sperm samples were incubated with anti-ZP2 IgG or with anti-ZP3 Fab fragments. When capacitated sperm were treated successively, with (a) ZP3 glycopeptides, (b) anti-ZP3 Fab fragments, and (c) goat anti-rabbit IgG, ARs occurred in the majority of sperm. An alternative approach to examine this hypothesis used ZP proteins obtained from tubal eggs treated previously with bioactive phorbol diester (12-O-tetradecanoyl phorbol-13-acetate [TPA]). This preparation arrests capacitated sperm in an intermediate state of the AR. We demonstrate here that these sperm can be induced to undergo a complete AR by subsequent treatment with anti-ZP3 IgG. Together, these findings are consistent with the hypothesis under examination, and suggest that the aggregation of sperm molecules recognized by ZP3 glycopeptides or by TPA-treated ZP is sufficient to trigger the events that occur during acrosomal exocytosis. PMID- 2544603 TI - Isolation and characterization of membranes from bovine liver which are highly enriched in mannose 6-phosphate receptors. AB - We have developed a method for the isolation of the subcellular organelles from bovine liver which are enriched in the cation-independent mannose 6-phosphate receptor (CI-MPR) and the cation-dependent mannose 6-phosphate receptor (CD-MPR). The purification scheme consists of sedimentation of a postnuclear supernatant fraction on a sucrose gradient followed by immunoisolation using specific anti peptide antibodies conjugated to magnetic polystyrene beads. Antibodies that recognize the cytoplasmic domain of either the CI-MPR or the CD-MPR routinely give membrane preparations that are approximately 50-fold enriched in each of the respective receptors, as determined by quantitative Western blotting. The immunoisolated membranes are also enriched in the other MPR, as well as in the asialoglycoprotein receptor. They contain significantly lower levels of enzyme activities representative of the plasma membrane (5' nucleotidase) or the Golgi complex (galactosyltransferase and sialyltransferase). There is little or no enrichment for either the lysosomal enzymes beta-hexosaminidase and tartrate resistant acid phosphatase, or the mitochondrial enzyme succinate-tetrazolium reductase. These data, together with electron microscopy of the immunoisolated material, suggest that the bulk of MPR-containing membranes we have isolated from bovine liver correspond to endosomes. Analysis by SDS-PAGE indicates that several proteins, including two with apparent molecular weights of 170 K and 400 K, are significantly enriched in the purified fractions and may represent potential markers for MPR-containing endosomes. PMID- 2544602 TI - Relations between the intracellular pathways of the receptors for transferrin, asialoglycoprotein, and mannose 6-phosphate in human hepatoma cells. AB - We compared the intracellular pathways of the transferrin receptor (TfR) with those of the asialoglycoprotein receptor (ASGPR) and the cation-independent mannose 6-phosphate receptor (MPR)/insulin-like growth factor II receptor during endocytosis in Hep G2 cells. Cells were allowed to endocytose a conjugate of horseradish peroxidase and transferrin (Tf/HRP) via the TfR system. Postnuclear supernatants of homogenized cells were incubated with 3,3'-diaminobenzidine (DAB) and H2O2. Peroxidase-catalyzed oxidation of DAB within Tf/HRP-containing endosomes cross-linked their contents to DAB polymer. The cross-linking efficiency was dependent on the intravesicular Tf/HRP concentration. The loss of detectable receptors from samples of cell homogenates treated with DAB/H2O2 was used as a measure of colocalization with Tf/HRP. To compare the distribution of internalized plasma membrane receptors with Tf/HRP, cells were first surface labeled with 125I at 0 degrees C. After uptake of surface 125I-labeled receptors at 37 degrees C in the presence of Tf/HRP, proteinase K was used at 0 degrees C to remove receptors remaining at the plasma membrane. Endocytosed receptors were isolated by means of immunoprecipitation. 125I-TfR and 125I-ASGPR were not sorted from endocytosed Tf/HRP. 125I-MPR initially also resided in Tf/HRP-containing compartments, however 70% was sorted from the Tf/HRP pathway between 20 and 45 min after uptake. To study the accessibility of total intracellular receptor pools to endocytosed Tf/HRP, nonlabeled cells were used, and the receptors were detected by means of Western blotting. The entire intracellular TfR population, but only 70 and 50% of ASGPR and MPR, respectively, were accessible to endocytosed Tf/HRP. These steady-state levels were reached by 10 min of continuous Tf/HRP uptake at 37 degrees C. We conclude that 30% of the intracellular ASGPR pool is not involved in endocytosis (i.e., is silent). Double labeling immunoelectron microscopy on DAB-labeled cells showed a considerable pool of ASGPR in secretory albumin-positive, Tf/HRP-negative, trans-Golgi reticulum. We suggest that this pool represents the silent ASGPR that has been biochemically determined. A model of receptor transport routes is presented and discussed. PMID- 2544605 TI - Biogenesis of peroxisomes: immunocytochemical investigation of peroxisomal membrane proteins in proliferating rat liver peroxisomes and in catalase-negative membrane loops. AB - Treatment of rats with a new hypocholesterolemic drug BM 15766 induces proliferation of peroxisomes in pericentral regions of the liver lobule with distinct alterations of the peroxisomal membrane (Baumgart, E., K. Stegmeier, F. H. Schmidt, and H. D. Fahimi. 1987. Lab. Invest. 56:554-564). We have used ultrastructural cytochemistry in conjunction with immunoblotting and immunoelectron microscopy to investigate the effects of this drug on peroxisomal membranes. Highly purified peroxisomal fractions were obtained by Metrizamide gradient centrifugation from control and treated rats. Immunoblots prepared from such peroxisomal fractions incubated with antibodies to 22-, 26-, and 70-kD peroxisomal membrane proteins revealed that the treatment with BM 15766 induced only the 70-kD protein. In sections of normal liver embedded in Lowicryl K4M, all three membrane proteins of peroxisomes could be localized by the postembedding technique. The strongest labeling was obtained with the 22-kD antibody followed by the 70-kD and 26-kD antibodies. In treated animals, double-membraned loops with negative catalase reaction in their lumen, resembling smooth endoplasmic reticulum segments as well as myelin-like figures, were noted in the proximity of some peroxisomes. Serial sectioning revealed that the loops seen at some distance from peroxisomes in the cytoplasm were always continuous with the peroxisomal membranes. The double-membraned loops were consistently negative for glucose-6 phosphatase, a marker for endoplasmic reticulum, but were distinctly labeled with antibodies to peroxisomal membrane proteins. Our observations indicate that these membranous structures are part of the peroxisomal membrane system. They could provide a membrane reservoir for the proliferation of peroxisomes and the expansion of this intracellular compartment. PMID- 2544606 TI - Human neuroblastoma cells acquire regulated secretory properties and different sensitivity to Ca2+ and alpha-latrotoxin after exposure to differentiating agents. AB - IMR-32 human neuroblastoma cells are unable to release [3H]dopamine in response to secretagogues. However, they express a normal complement of membrane receptors and ion channels which are efficiently coupled to second messenger production. In the present study we took advantage of the ability of this cell line to differentiate in vitro in the presence of either dibutyrryl-cAMP or 5 bromodeoxyuridine, to analyze any developmentally regulated changes in its secretory properties. Uptake, storage, and release of [3H]dopamine were studied biochemically and by autoradiography. The calcium ionophore ionomycin, phorbol 12 myristate 13-acetate and the presynaptic acting neurotoxin alpha-latrotoxin were used in both control and differentiated cells as secretagogue agents. The presence of secretory organelles was investigated by electron microscopy; the expression of secretory organelle markers, such as chromogranin/secretogranin proteins (secretory proteins) and synaptophysin (membrane protein), was detected by Western blotting and immunofluorescence. The results obtained indicate that IMR-32 cells acquire regulated secretory properties after in vitro drug-induced differentiation: (a) they assemble "de novo" secretory organelles, as revealed by electron microscopy and detection of secretory organelle markers, and (b) they are able to store [3H]dopamine and to release the neurotransmitter in response to secretagogue stimuli. Furthermore, secretagogue sensitivity was found to be different, depending on the differentiating agent. In fact, dibutyrryl-cAMP treated cells release [3H]dopamine in response to alpha-latrotoxin, but not in response to ionomycin, whereas 5-bromodeoxyuridine treated cells release the neurotransmitter in response to both secretagogues. All together these results suggest that IMR-32 cells represent an adequate model for studying the development of the secretory apparatus in cultured human neurons. PMID- 2544607 TI - Interactions of cytoplasmic granules with microtubules in human neutrophils. AB - Ultrastructural and functional studies of degranulation responses by human neutrophils have suggested that microtubules (MTs) have a role in the intracellular transport of neutrophil granules. We have found that granule-MT complexes can be isolated from disrupted taxol-treated (1.0 microM) neutrophils, visualized by electron microscopy, and quantified in terms of granules per MT length. After incubation of neutrophils with the chemotactic peptide N-formyl methionyl-leucyl-phenylalanine (fMLP), granule-MT complex formation was found to be increased two- to threefold. Enhanced binding of granules to MTs was detectable within 30 s of fMLP stimulation and was dependent on the concentration of fMLP. Incubation of cells with dibutyryl cAMP inhibited this fMLP-stimulated granule-MT complex formation in a dose-responsive fashion. These granule-MT interactions could be reproduced in a cell-free system with neutrophil granules isolated by density gradient centrifugation and MTs polymerized from phosphocellulose-purified tubulin. Furthermore, reconstituted granule-MT interactions were found to be modulated by ATPase inhibitors. Sodium orthovanadate increased granule-MT interactions in a concentration-dependent manner, while AMP-PNP, a nonhydrolyzable ATP analogue, and N-ethylmaleimide decreased or eliminated these interactions. In addition, we found that a MT activated ATPase could be recovered from intact neutrophil granules by salt extraction, and that extracts enriched in this ATPase contained a polypeptide of between 115 and 120 kD which binds ATP and is immunologically related to kinesin. These studies demonstrate that cytoplasmic granules interact with MTs in human neutrophils in a regulated stimulus-responsive manner, and they suggest that such interactions may involve an MT-based, ATPase-dependent, vesicle translocation system as has been demonstrated in other types of cells. PMID- 2544608 TI - Expression of fibroblast growth factor by F9 teratocarcinoma cells as a function of differentiation. AB - F9 teratocarcinoma stem cells treated with retinoic acid (RA) and dibutyryl cAMP (but2 cAMP) differentiate into embryonic parietal endoderm. Using heparin affinity chromatography, endothelial cell proliferation assays, immunoprecipitation, and Western analysis with antibodies specific for acidic and basic fibroblast growth factors (FGFs), we detected biologically active FGF in F9 cells only after differentiation. A bovine basic FGF cDNA probe hybridized to 2.2 kb mRNAs in both F9 stem and parietal endoderm cells and to a 3.8-kb mRNA in F9 stem cells. A genomic DNA probe for acidic FGF hybridized to a 5.8-6.0-kb mRNA in both F9 stem and parietal endoderm cells, and to a 6.0-6.3-kb mRNA only in parietal endoderm cells. Although these FGF mRNAs were present in the stem cells, we could find no evidence that F9 stem cells synthesized FGFs, whereas differentiated F9 cells synthesized both acidic and basic FGF-like proteins. We conclude that biologically active factors with properties characteristic of acidic and basic FGF are expressed by F9 parietal endoderm cells after differentiation. Differentiating embryonic parietal endoderm thus may serve as a source of FGF molecules in the developing blastocyst, where these factors appear to play a central role in subsequent embryogenesis. PMID- 2544609 TI - Actions of insulin-like growth factor-I on the B104 neuronal cell line: effects on cell replication, receptor characteristics, and influence of secreted binding protein on ligand binding. AB - Several peptide growth factors influence the growth and differentiation of neural cells. To investigate further the growth-promoting effects of the somatomedins on cells of neural origin, the authors characterized the binding and mitogenic effects of insulin-like growth factor-I (IGF-I) on a functionally differentiated rat neuronal cell line (B104). Specific, high-affinity (Kd approximately equal to 10(-9) M) receptors for IGF-I were abundant (approximately 124,000 binding sites/B104 cell). These IGF-I receptors were similar to those of non-neural tissue in that they contained 135,000 dalton binding subunits (demonstrated by affinity labeling and autoradiography) and recognized insulin at high concentrations. IGF-I was more potent than insulin at stimulating B104 cell replication in serum-free medium and, at an initial concentration of 100 ng/ml, was the only exogenous growth factor needed to maintain growth through several cell divisions. Furthermore, cells of later passage were found to secrete specific IGF binding proteins that produced an unusual, biphasic binding curve in radioligand displacement studies. These binding proteins apparently sequester IGF I, limiting its access to the cell. Experiments with B104 cells may provide useful information about the role of IGFs and their binding proteins as potential regulators of growth and differentiation of the primitive neuroblast. PMID- 2544610 TI - Epidermal growth factor stimulation of prostacyclin production by cultured aortic smooth muscle cells: requirement for increased cellular calcium levels. AB - We have examined the ability of epidermal growth factor (EGF) to regulate prostacyclin production by cultured A10 smooth muscle cells. EGF by itself had no effect on prostacyclin production, but it augmented the response to arg8 vasopressin. An AGF stimulation of prostacyclin production was also observed in the presence of the calcium ionophore A23187; it therefore seemed likely that the key event required for EGF to stimulate prostacyclin production might be an increase in the available cellular Ca2+. Studies with 45Ca2+ showed that vasopressin both mobilised Ca2+ from intracellular stores and increased the influx of extracellular Ca2+ into A10 cells. The increase in prostacyclin production caused by vasopressin and the augmentation by EGF were both abolished by TMB-8, an antagonist of Ca2+ mobilisation, by EGTA, a chelator of Ca2+ ions, or by incubating cultures in the absence of added Ca2+. These results were consistent with a central role for Ca2+ in the responses and showed that both intracellular and extracellular sources of Ca2+ were important for the triggering of prostacyclin production. The increases in prostacyclin production were only marginally affected by nifedipine, and no responses were seen (either in the absence or presence of EGF) when KCl was used to depolarise the cell membrane. These data indicated that uptake of Ca2+ ions via voltage-dependent channels was unlikely to be a major factor in the stimulation of prostanoid production. We conclude that the ability of EGF to stimulate prostacyclin production in A10 smooth muscle cells depends upon a concurrent stimulus that will increase available intracellular Ca2+ levels. PMID- 2544613 TI - Regulation of the sodium-potassium pump in cultured rat skeletal myotubes by intracellular sodium ions. AB - The properties of the Na-K pump and some of the factors controlling its amount and function were studied in rat myotubes in culture. The number of Na-K pump sites was quantified by measuring the amount of [3H]ouabain bound to whole-cell preparations. Activity of the pump was determined by measurement of ouabain sensitive 86Rb-uptake and component of membrane potential. Chronic treatment of myotubes with tetrodotoxin (TTX), which lowers [Na]i, decreased the number of Na K pumps, the ouabain-sensitive 86Rb uptake, and the size of the electrogenic pump component of Em. In contrast, chronic treatment with either ouabain or veratridine, which increases [Na+]i, resulted in an elevated level of Na-K pump sites. This effect was blocked by inhibitors of protein synthesis. Neither rates of degradation nor affinity of pump sites in cells treated with TTX, veratridine, or ouabain differred from those in control cells. The number and activity of Na-K pump sites were unaffected by chronic elevation in [Ca]i or chronic depolarization. We conclude that alterations in the level in intracellular Na ions play the major role in regulation of Na-K pump synthesis in cultured mammalian skeletal muscle. PMID- 2544614 TI - Characterization of the receptor for insulin-like growth factor II in bone cells. AB - We have previously shown that insulin-like growth factor II (IGF-II) is produced by bone cells and that IGF-II stimulates cell proliferation and collagen synthesis in bone cells. We now extend these in vitro findings by demonstrating specific IGF-II binding to bone cells derived from newborn mouse calvaria and embryonic chick calvaria. The kinetics of [125I] IGF-II binding in embryonic chick calvaria cells showed time and temperature dependence. Scatchard analysis of [125I]IGF-II binding to chick calvaria cells showed an apparent Kd of 1.4 x 10(-10) M, with a calculated receptor site concentration of 40,000/cell. The specificity characteristics showed that IGF-II was significantly more potent than IGF-I or insulin in displacing IGF-II tracer. Competition for binding of [125I]IGF-II by unlabeled IGF-II showed a dose-dependent displacement between 0.5 and 25 ng/ml. Fifty percent displacement of [125I]IGF-II binding to chick and mouse calvarial cells was achieved at 1-2 ng/ml; 90% of specific binding of [125I]IGF-II was displaceable in the presence of 125 ng/ml of unlabeled IGF-II. IGF-I showed less than 5% cross reactivity for displacement of [125I]IGF-II binding to chick and mouse bone cells. Type II receptor inhibitory antibodies, R II-PAB1 inhibited the binding of [125I]IGF-II to mouse bone cells and H-35 rat hepatoma cells (which contain type II but not type I receptors) in a dose dependent manner. R-II-PAB1 also inhibited basal cell proliferation as well as IGF-II-, IGF-I-, and fibroblast growth factor (FGF)-induced cell proliferation in mouse bone cells. In chick calvaria bone cells and TE89 human osteosarcoma cells, R-II-PABI inhibited neither binding of [125I]IGF-II nor IGF-II-induced cell proliferation. These results together with our findings that IGF-II increased chick bone cell proliferation in the presence of maximal doses of IGF-I suggest that at least part of the mitogenic action of IGF-II is mediated through type II rather than type I receptors in bone cells. PMID- 2544611 TI - Electrophysiological properties of isolated rat liver cells. AB - The electrophysiological properties of isolated rat liver cells were studied using the patch clamp method in whole-cell configuration. The membrane potential in isolated hepatocytes was -42 +/- 7 mV (n = 20). The input resistance (Rin) and the time constant (tau m) were 51 +/- 17 M (the range of 34 to 180 M omega) (n = 20) and 4.2 +/- 1.0 msec (the range of 3 to 16.5 ms) (n = 20). Assuming that the specific membrane capacitance is 1 microF/cm2, the membrane resistance and membrane capacitance were 42. +/- 9.0 K omega cm2 and 87 +/- 27 pF. These values indicate that isolated rat hepatocytes are not abnormally permeable or leaky. The current-voltage relationship was linear with no rectification. The depolarizing pulse from the resting potential did not induce fast or slow inward currents even when norepinephrine or high Ca2 (3.6 mM) were applied. This indicates that there is no voltage-sensitive Ca2+ channel in the isolated hepatocytes. PMID- 2544612 TI - Bone marrow-derived stromal cell line expressing osteoblastic phenotype in vitro and osteogenic capacity in vivo. AB - Marrow stroma has been shown to have osteogenic potential. Here we report the characterization of a unique stromal cell line derived from mouse bone marrow (MBA-15), which expresses osteoblastic phenotype in vitro and forms bone in vivo. More than 70% of cells in culture were histochemically positive for alkaline phosphatase. The enzyme levels were enhanced threefold when cultures were treated with dexamethasone. Gel electrophoresis of [3H]-proline-labeled cultures showed that MBA-15 cells produced only type I collagen. These cells were responsive to PTH, as indicated by a 50-fold increase in intracellular cAMP. Prostaglandin E2, but not calcitonin, stimulated cAMP up to 70-fold. When cultures were grown to confluence and fed daily with ascorbic acid and beta-glycerophosphate, the cells formed a Von Kossa positive, thick extracellular matrix, shown to contain hydroxyapatite crystals. MBA-15 cells produced mineralized bone when implanted in diffusion chambers. These results indicate that the MBA-15 cell line possesses osteoblastic features in vitro and osteogenic capacity in vivo. PMID- 2544615 TI - Reduction of adenine nucleotide content of clone 4 MDCK cells: effects on multiplication, protein synthesis, and morphology. AB - The antitumor agent hadacidin (N-formyl-hydroxyamino-acetic acid), at 4 mM, inhibited the multiplication of clone 4 Madin Darby canine kidney (MDCK) cells within 24 hr. Growth resumed rapidly upon replacement of hadacidin with aspartate, an observation consistent with the drug's action as a competitive inhibitor of adenylosuccinate synthetase, an enzyme in adenine nucleotide biosynthesis. Data indicate that the drug-treated cells were arrested in S phase of the cell cycle. Accompanying inhibition of multiplication was a 16-fold increase in the area occupied by the cells and a refractoriness to release by treatment with trypsin. None of these changes occurred when 0.5 mM adenosine was included in the incubation mixture containing the inhibitor. Hadacidin decreased the adenosine triphosphate (ATP) and cyclic adenosine monophosphate (cAMP) content of the cells as well as the rate at which 3H-leucine was incorporated into protein. In the presence of 1 mM dibutyryl cAMP and theophylline, the drug had no effect on cell division and protein synthesis. The data suggest that, in clone 4 MDCK cells, the effects of hadacidin are mediated by diminishing the level of cAMP. PMID- 2544616 TI - Regulation of beta-adrenergic receptor expression in rat liver. AB - To begin defining the factors regulating neurotransmitter receptor expression, we examined beta-adrenergic receptors in rat liver in vivo and in primary liver cultures under defined hormonal conditions. beta-receptors described a remarkable developmental profile in vivo, increasing fivefold between embryonic days 16 and 20, and decreasing tenfold by early adulthood. The developmental decrease reflected reduced receptor number without a change in receptor properties. The ontogenetic decrease appeared to be specific for beta-receptors; alpha-receptors developed in a hyperbolic fashion, reaching high plateau values by the third postnatal week. Adult rat liver cells plated into culture re-expressed high beta receptor levels, exhibiting a 4-8-fold increase. A similar pattern of expression of the beta-receptors, having similar pharmacological properties, was observed in primary liver cultures maintained in serum-free medium, in a serum-supplemented medium or in several variations of a serum-free, hormonally defined medium designed for primary liver cultures. Thus, the degree of expression of the beta receptors was not found affected by various hormones, by serum, or by any medium condition. By contrast, the degree of expression of the beta-receptors was markedly sensitive to cell density. High expression of the beta-receptors was observed at low cell densities (1-3 x 10(6) cells/150 mm dish), and low expression or no expression was observed in confluent cultures (10-20 x 10(6) cells/150 mm dish). Our experiments suggest that beta-receptor expression does not follow an immutable program, but may be regulated by density-dependent cell cell interactions. PMID- 2544619 TI - High-performance liquid chromatographic determination of citrinin in cereals using an acid-buffered silica gel column. PMID- 2544617 TI - Dexamethasone effects on creatine kinase activity and insulin-like growth factor receptors in cultured muscle cells. AB - We examined the effects of dexamethasone on creatine kinase (CK) activity and insulin-like growth factor I (IGF-I) binding in two skeletal muscle-derived cell lines (mouse, C2C12; rat, L6) and in one cardiac muscle-derived cell line (rat, H9c2). Dexamethasone treatment during differentiation of cultured cells caused a dose-dependent increase in CK activity as well as an increase in the degree of myotube formation in C2C12 and L6, whereas H9c2 cells did not exhibit significant CK activities during culture or dexamethasone treatment. Dexamethasone treatment of C2C12 did not stimulate proliferation in differentiating cultures, but a dose dependent increase in the number of nuclei was observed for L6 concomitant with increased CK activity. In L6 the increased CK activity may therefore reflect a dose-dependent increase in proliferation. Short-term (48 hr) treatment of C2C12 with dexamethasone (20 nM) did not appear to alter myoblast fusion but reversibly increased CK activity. In C2C12 the observed increase in CK, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activities with dexamethasone treatment suggest modulation of protein expression and/or turnover. Although the data for dexamethasone effects on CK activities varied in each of the cell lines, consistent behavior was observed in all three cell lines when IGF I binding was examined. IGF-I binding to dexamethasone-treated cells (50 nM for 24 hr the day prior to confluence) resulted in an increased number of available binding sites, with no effect on the binding affinities. Affinity cross linking and autoradiography indicated that the increase in IGF-I binding was the result of dexamethasone up-regulation of type I IGF receptors. Our data for all three muscle cell lines suggest that similar heterologous hormone receptor modulation of type I IGF receptor sites occurs with dexamethasone treatment. PMID- 2544618 TI - Protein kinase C modulates effects of prostanoids on cyclic adenosine monophosphate in guinea pig chief cells. AB - In the course of examining the role of protein kinase C in signal transduction in dispersed chief cells from guinea pig stomach, we observed that phorbol esters inhibit prostaglandin (PG)-stimulated increases in cyclic adenosine monophosphate (cAMP). Phorbol 12-myristate 13-acetate (PMA), an activator of protein kinase C, decreased maximal levels of PGE2-stimulated cAMP by 40%. This dose-dependent effect was observed within 30 sec and was maximal by 1 min of incubation at 37 degrees C. Phorbols that do not activate protein kinase C did not have this effect. Adding H7, a protein kinase C inhibitor, abolished the inhibitory effects of PMA, indicating that these effects are not caused by activation of cyclic nucleotide phosphodiesterases. PMA did not alter the increase in cellular cAMP caused by cholera toxin, forskolin, secretin, or vasoactive intestinal peptide. Hence the site of these prostanoid-specific actions of protein kinase C does not appear to be stimulatory or inhibitory guanine nucleotide binding proteins or the catalytic component of the adenylyl cyclase system. In dispersed chief cells, activation of protein kinase C may inhibit prostanoid-induced stimulation of the adenylyl cyclase system by a direct effect on prostaglandin receptors. PMID- 2544620 TI - Complementary information from isotachophoresis and high-performance liquid chromatography in peptide analysis. AB - Reversed-phase high-performance liquid chromatography is a valuable analytical technique to support the synthesis, isolation and purification of peptides, as is illustrated by some critical separations. In addition to this technique, capillary isotachophoresis can give useful information on the purity determination of peptides and on the presence of ionic compounds of a non peptidic nature. With regard to the latter aspect, isotachophoresis proved to be a suitable technique as a check on the effective removal of salts after preparative high-performance liquid chromatography. PMID- 2544622 TI - Chronic blockade of endogenous atrial natriuretic polypeptide (ANP) by monoclonal antibody against ANP accelerates the development of hypertension in spontaneously hypertensive and deoxycorticosterone acetate-salt-hypertensive rats. AB - To explain the pathophysiological significance of endogenous atrial natriuretic polypeptide (ANP) in the development of hypertension, we examined the effect of chronic, repetitive administrations of MAb raised against alpha-rat ANP in two rat models of hypertension, spontaneously hypertensive rats of the stroke prone substrain (SHR-SP), and deoxycorticosterone acetate (DOCA)-salt rats. Weekly intravenous administrations of MAb with high affinity for alpha-rat ANP, named KY ANP-II (MAb[KY-ANP-II]), started at the age of 6 wk, significantly augmented the rise in blood pressure of SHR-SP, compared with control SHR-SP treated with another MAb with quite low affinity for alpha-rat ANP, named KY-ANP-I (MAb[KY-ANP I]), throughout the observation period. The administrations of MAb[KY-ANP-II] had no significant effect on blood pressure of age-matched normotensive Wistar Kyoto rats, compared with those receiving MAb[KY-ANP-I]. Weekly administrations of MAb[KY-ANP-II] also significantly aggravated hypertension in DOCA-salt rats. Blood pressure of DOCA-salt rats treated with MAb[KY-ANP-II] was significantly higher than that of DOCA-salt rats treated with MAb[KY-ANP-I] throughout 8 wk of DOCA and 1% saline administration. The administration of MAb[KY-ANP-II] also significantly attenuated exaggerated diuresis and natriuresis in DOCA-salt rats compared with those treated with MAb[KY-ANP-I]. Elevated plasma cGMP levels of both SHR-SP and DOCA-salt rats were significantly reduced by the administration of MAb[KY-ANP-II]. These results suggest the compensatory role of augmented secretion of ANP in these hypertensive rats and support the concept that augmented secretion of ANP could represent an antihypertensive deterrent mechanism. PMID- 2544621 TI - Complement-mediated phagocytosis of herpes simplex virus by granulocytes. Binding or ingestion. AB - The role of complement receptors in phagocytosis of herpes simplex virus (HSV) by PMN was examined. Complement components were deposited on the surface of the virus particle in the presence or absence of specific anti-HSV antibodies. Flow cytometry was used to analyze the phagocytosis of fluorescence-labeled viruses and demonstrated that although a virion is able to associate with PMN in the presence of complement alone, the granulocyte is not triggered to mount a metabolic burst. Efficient stimulation of PMN occurs when complexes are formed consisting of virus, specific antibodies, and complement. To address the question whether the viruses were inside or outside the cell, a combined enhancement/quenching method was developed using ammonium chloride as a lysosomotropic agent and trypan blue as a quenching dye. The data indicate that Fc receptor-mediated phagocytosis by PMN results in the ingestion of all cell associated herpes virions. Interactions of virions through PMN-complement receptors CR1 and CR3 results solely in binding to the PMN but not in internalization. Interactions via both complement and Fc receptors cause synergistic stimulation of the PMN and result in very efficient association of viruses, greater than 80% of which were inside the cell. PMID- 2544624 TI - Insertion of the v-Ha-ras oncogene induces differentiation of calcitonin producing human small cell lung cancer. AB - Human small cell lung cancers (SCLC) and cell lines derived therefrom are phenotypically heterogeneous concerning neuroendocrine differentiation. Unlike most SCLC tumors and cell lines that express poorly differentiated neuroendocrine phenotypes, the SCLC cell line DMS 53 exhibits mature endocrine differentiation features, including unusually high expression of the gene for the peptide hormone, calcitonin (CT). We now report that introduction of the viral Harvey ras (v-rasH) oncogene into DMS 53 cells via retroviral infection, with resultant constitutive expression, results in increased features of neuroendocrine differentiation. 7-10 d after infection the cells demonstrated altered morphology, increased CT secretion, increased CT gene expression, markedly diminished cellular proliferation, and nearly abolished methylcellulose cloning efficiency. This response of DMS 53 cells to v-rasH is unlike the tumor progression effects we have previously observed in other SCLC lines. Significantly, the differentiation response that follows expression of the virally introduced v-rasH oncogene in DMS 53 cells is similar to that of neoplastic neuroendocrine cell lines derived from adrenal pheochromocytes and thyroid C cells. The effects of constitutive v-rasH expression in DMS 53 SCLC cells and other neuroendocrine cell lines suggest an important role for rasH or related genes in neuroendocrine differentiation. PMID- 2544623 TI - Exercise intolerance, lactic acidosis, and abnormal cardiopulmonary regulation in exercise associated with adult skeletal muscle cytochrome c oxidase deficiency. AB - A 27-yr-old woman with lifelong severe exercise intolerance manifested by muscle fatigue, lactic acidosis, and prominent symptoms of dyspnea and tachycardia induced by trivial exercise was found to have a skeletal muscle respiratory chain defect characterized by low levels of reducible cytochromes a + a3 and b in muscle mitochondria and marked deficiency of cytochrome c oxidase (complex IV) as assessed biochemically and immunologically. Investigation of the pathophysiology of the exercise response in the patient revealed low maximal oxygen uptake (1/3 that of normal sedentary women) in cycle exercise and impaired muscle oxygen extraction as indicated by profoundly low maximal systemic arteriovenous oxygen difference (5.8 ml/dl; controls = 15.4 +/- 1.4, mean +/- SD). The increases in cardiac output and ventilation during exercise, normally closely coupled to muscle metabolic rate, were markedly exaggerated (more than two- to threefold normal) relative to oxygen uptake and carbon dioxide production accounting for prominent tachycardia and dyspnea at low workloads. Symptoms in our patient are similar to those reported in other human skeletal muscle respiratory chain defects involving complexes I and III, and the exaggerated circulatory response resembles that seen during experimental inhibition of the mitochondrial respiratory chain. These results suggest that impaired oxidative phosphorylation in working muscle disrupts the normal regulation of cardiac output and ventilation relative to muscle metabolic rate in exercise. PMID- 2544625 TI - Decreased bioactivity of the guanine nucleotide-binding protein that stimulates adenylate cyclase in hearts from cardiomyopathic Syrian hamsters. AB - We investigated regulation of cardiac adenylate cyclase in 29-d-old BIO 14.6 Syrian hamsters, which inherit cardiomyopathy as an autosomal recessive trait. Pharmacologic stimulation of adenylate cyclase in cardiac membranes with isoproterenol, fluoride ion, guanine nucleotide, forskolin, and manganous ion indicated that there was defective coupling of the guanine nucleotide-binding protein that stimulates adenylate cyclase (Gs) to adenylate cyclase. Cyc complementation assays revealed congruent to 50% less Gs activity in cardiac and skeletal muscle from cardiomyopathic hamsters. Despite this decrease in functional Gs, there were no changes in immunologic levels of the alpha-subunit of Gs (alpha Gs) or in levels of mRNA encoding alpha Gs. The defect in Gs bioactivity was limited to cardiac and skeletal muscle, occurred only in animals homozygous for the dystrophic trait, and was demonstrable before any cardiac abnormalities were evident on light microscopy. By contrast, cardiac levels of beta-adrenergic receptors were not different in cardiac membranes from BIO 14.6 hamsters. We conclude that a functional defect in alpha Gs may contribute to a contractile abnormalities in the cardiomyopathic BIO 14.6 hamster. However, the etiology of the alpha Gs defect remains obscure. PMID- 2544626 TI - Bicarbonate-dependent and -independent intracellular pH regulatory mechanisms in rat hepatocytes. Evidence for Na+-HCO3- cotransport. AB - Using the pH-sensitive dye 2,7-bis(carboxyethyl)-5(6)-carboxy-fluorescein and a continuously perfused subconfluent hepatocyte monolayer cell culture system, we studied rat hepatocyte intracellular pH (pHi) regulation in the presence (+HCO3-) and absence (-HCO3-) of bicarbonate. Baseline pHi was higher (7.28 +/- 09) in +HCO3- than in -HCO3- (7.16 +/- 0.14). Blocking Na+/H+ exchange with amiloride had no effect on pHi in +HCO3- but caused reversible 0.1-0.2-U acidification in HCO3- or in +HCO3- after preincubation in the anion transport inhibitor 4,4' diisothiocyano-2,2'-disulfonic acid stilbene (DIDS). Acute Na+ replacement in +HCO3- alos caused acidification which was amiloride independent but DIDS inhibitible. The recovery of pHi from an intracellular acid load (maximum H+ efflux rate) was 50% higher in +HCO3- than in -HCO3-. Amiloride inhibited H+ effluxmax by 75% in -HCO3- but by only 27% in +HCO3-. The amiloride-independent pHi recovery in +HCO3- was inhibited 50-63% by DIDS and 79% by Na+ replacement but was unaffected by depletion of intracellular Cl-, suggesting that Cl-/HCO3- exchange is not involved. Depolarization of hepatocytes (raising external K+ from 5 to 25 mM) caused reversible 0.05-0.1-U alkalinization, which, however, was neither Na+ nor HCO3- dependent, nor DIDS inhibitible, findings consistent with electroneutral HCO3- transport. We conclude that Na+-HCO3- cotransport, in addition to Na+/H+ exchange, is an important regulator of pHi in rat hepatocytes. PMID- 2544627 TI - Expression of diverse Na+ channel messenger RNAs in rat myocardium. Evidence for a cardiac-specific Na+ channel. AB - This study examined the diversity of Na+ channel gene expression in intact cardiac tissue and purified myocardial cells. The screening of neonatal rat myocardial cell cDNA libraries with a conserved rat brain Na+ channel cDNA probe, resulted in the isolation and characterization of a putative rat cardiac Na+ channel cDNA probe (pCSC-1). The deduced amino acid sequence of pCSC-1 displayed a striking degree of homology with the eel, rat brain-1, and rat brain-2 Na+ channel, thereby identifying pCSC-1 as a related member of the family of Na+ channel genes. Northern blot analysis revealed the expression of a 7-kb CSC-1 transcript in rat cardiac tissue and purified myocardial cells, but little or no detectable expression of CSC-1 in rat brain, skeletal muscle, denervated skeletal muscle, or liver. Using RNase protection and Northern blot hybridization with specific rat brain Na+ channel gene probes, expression of the rat brain-1 Na+ channel was observed in rat myocardium, but no detectable expression of the rat brain-2 gene was found. This study provides evidence for the expression of diverse Na+ channel mRNAs in rat myocardium and presents the initial characterization of a new, related member of the family of Na+ channel genes, which appears to be expressed in a cardiac-specific manner. PMID- 2544628 TI - Expression of multiple Na+,K+-adenosine triphosphatase isoform genes in human hematopoietic cells. Behavior of the novel A3 isoform during induced maturation of HL60 cells. AB - Multiple isoenzymes of the Na+,K+-ATPase (alpha, alpha+, and alpha 3) have been identified by molecular cloning (Shull, G. E., J. Greeb, and J. B. Lingrel. 1986. Biochemistry. 25:8125-8132; and Schneider, J. W., R. W. Mercer, and E. J. Benz, Jr. 1987. Clin. Res. 35:585A. [Abstr.]). At least one of these, the alpha 3 chain, represents a novel form for which protein products and enzymatic activities are just beginning to be defined in rodents. We have recently demonstrated that expression of alpha 3 is largely confined to neuromuscular tissues of fetal and adult rats (Schneider, J. W., R. W. Mercer, M. Gilmore Hebert, M. F. Utset, C. Lai, A. Greene, and E. J. Benz, Jr. 1988. Proc. Natl. Acad. Sci. USA. 85:284-288). We now report that certain human leukemia cell lines including HL60, HEL, and Molt 4 express mRNA for both alpha and alpha 3 isoforms of Na+,K+-ATPase; mRNA was not detected in several other cell lines, including K562 and U937; no cell lines expressed alpha+ mRNA. In uninduced HL60 cells, alpha 3 mRNA comprised 20-30% of total Na+,K+-ATPase mRNA. Furthermore, in HL60 and HEL cells, both alpha and alpha 3 mRNA declined after induction of maturation by DMSO, retinoic acid, or hemin. However, the reduction in alpha 3 mRNA was far more dramatic. alpha 3 mRNA virtually disappeared, but alpha mRNA declined by only approximately 50%. In contrast, when maturation of HL60 cells along the monocyte/macrophage lineage was induced by exposure to phorbol esters, alpha 3 mRNA remained abundant. Moreover, mRNA for the beta subunit of the Na+,K+-ATPase increased dramatically. Our results demonstrate that the alpha 3 isoform, formerly thought to be confined to neuromuscular tissues, is expressed in restricted lineages of hematopoietic origin. These leukemia cell lines should provide a useful model for analyzing regulation of the alpha 3 isoform gene and characterization of alpha 3 isoform activities. PMID- 2544629 TI - Active proton secretion and potassium absorption in the rabbit outer medullary collecting duct. Functional evidence for proton-potassium-activated adenosine triphosphatase. AB - We examined the hypothesis that proton-potassium-activated adenosine triphosphatase (H-K-ATPase) mediates K absorption and acidification in the inner stripe of the outer medullary collecting duct (OMCDi). Rabbits were fed a low-K diet (0.55% K) for 7-14 d because we have demonstrated previously that this low-K diet stimulates K-absorptive flux by the OMCDi. Proton secretion was measured as net total CO2 flux (JTCO2) by microcalorimetry. After basal collections, either vehicle or an inhibitor of gastric H-K-ATPase, omeprazole (0.1 mM), was added to the perfusate during the second period. Addition of vehicle to the perfusate changed neither the transepithelial voltage (VT, in millivolts) nor the JTCO2. In contrast, the addition of omeprazole (0.1 mM) to the perfusate abolished JTCO2 (from 14.5 +/- 5.6 to -0.1 +/- 3.1 pmol.mm-1.min-1) without significantly affecting VT. In additional experiments, in 16 tubules there was significant net K absorption (JK) of 5.0 +/- 1.0 pmol.mm-1.min-1 during the basal period, which exceeded the rate of K absorption that could be attributed to a paracellular voltage-mediated pathway (JKP = 1.0 +/- 0.4 pmol.mm-1.min-1, P less than 0.01). Administration of vehicle did not significantly affect either VT or JK. However, omeprazole abolished JK (from 5.1 +/- 1.0 to 0.1 +/- 2.5 pmol.mm-1.min-1) without affecting VT or JNa. The present results demonstrate that the OMCDi possesses an active, omeprazole-sensitive acidification and K-absorptive mechanism. These findings are consistent with the presence of H-K-ATPase activity in this nephron segment. PMID- 2544630 TI - CD3-negative lymphoproliferative disease of granular lymphocytes containing Epstein-Barr viral DNA. AB - Lymphoproliferative disease of granular lymphocytes (LDGL) is a heterogeneous disorder and the pathogenesis is likely to be complex. Some patients with chronic active EBV (CAEBV) infection also have LDGL. To investigate the relationship between EBV infection and the pathogenesis of LDGL, we conducted a survey for EBV DNA sequences by Southern blot analysis of DNA obtained from the peripheral blood of seven patients with LDGL, including one with CAEBV infection. Interestingly, EBV DNA was detected in the sample from the patient with CAEBV infection, and in the samples from four other patients with CD3-LDGL. Moreover, a single band for the joined termini of the EBV genome was demonstrated in two samples, suggesting a clonal disorder of those LDGL. These findings strongly suggest that EBV may play a pathogenic role in some cases of LDGL. PMID- 2544631 TI - Angiotensin II stimulates early proximal bicarbonate absorption in the rat by decreasing cyclic adenosine monophosphate. AB - These studies explored the hypothesis that angiotensin II increases bicarbonate absorption in the proximal convoluted tubule (PCT) by decreasing intracellular cAMP. In vivo microperfusion was performed in rat PCT with measurements of bicarbonate absorption and of tubular fluid cAMP delivery, as a reflection of intracellular cAMP. Intravenous angiotensin II potently increased S1 PCT bicarbonate absorption (348 +/- 11 to 588 +/- 8 peq/min.min, P less than 0.001) and decreased tubular fluid cAMP (18 +/- 2 to 12 +/- 2 fmol/mm.min, P less than 0.05). Parathyroid hormone had the expected opposite effects, which were additive to those of angiotensin II. Over a wide range of hormonal activities, there was an excellent inverse relationship between hormonally modulated bicarbonate absorption and cAMP delivery. Pertussis toxin pretreatment significantly attenuated (by 35-45%) the angiotensin-induced increase in bicarbonate absorption and decrease in cAMP delivery, indicating Gi-protein intermediation. Luminal dibutyryl cAMP abolished the transport response to angiotensin II. In conclusion, these in vivo results suggest angiotensin II stimulates bicarbonate absorption in the S1 PCT by a G1-mediated depression in intracellular cAMP. PMID- 2544632 TI - Interphase cytogenetics using biotin and digoxigenin labelled probes I: relative sensitivity of both reporter molecules for detection of HPV16 in CaSki cells. AB - This study was undertaken to develop technology for the detection of nucleic acid using two different DNA probe reporter molecules, the ultimate aim being to differentially label two nucleic acids within the same nucleus. Digoxigenin and biotin were used to label DNA probes. The absolute and relative sensitivity of digoxigenin and biotin labelled DNA probes for detecting integrated human papilloma virus 16 (HPV16) was investigated in CaSki cells by non-isotopic in situ hybridisation (NISH). Several methods for the detection of labelled probes were also investigated. The optimal sensitivity of digoxigenin labelled probe was equivalent to that of biotin when alkaline phosphatase was used as the final detector. The median number of discrete viral signals discernible in each cell with the most sensitive detection system was seven to eight with both labelled probes. The average number of HPV16 genomes in each CaSki cell, derived by dot blot hybridisation, was about 270. The calculated absolute sensitivity of NISH for viral detection in this system is complex because of variation of signal size and number. Nevertheless, one signal per nucleus equates to as little as 30 to 40 viral copies, and probably much less. The ability to distinguish up to 15 discrete signals with both digoxigenin and biotin labelled probes in the nuclei of CaSki cells indicates that these methods will be useful in interphase cytogenetics in material routinely fixed in aldehyde. PMID- 2544633 TI - Interphase cytogenetics using biotin and digoxigenin labelled probes II: Simultaneous differential detection of human and papilloma virus nucleic acids in individual nuclei. AB - A method was developed for the simultaneous detection of viral and human DNA in contrasting colours in routine formalin fixed, paraffin wax embedded biopsy specimens. This was achieved by non-isotopic in situ hybridisation (NISH) with a biotinylated Y chromosome probe and digoxigenin labelled probe for human papilloma virus type 6 (HPV 6). The tissues studied were peripheral lymphocytes, tonsil, and penile warts. The hybridisation signals produced by biotinylated probes were visualised in red using streptavidin peroxidase and those produced by digoxigenin labelled probes as a blue/black colour using anti-digoxigenin alkaline phosphatase. In lymphocytes and tonsil 95-100% of cells had a detectable Y chromosome; in warts only 60-70% of infected keratinocytes near the skin surface had a demonstrable Y chromosome. This suggests that this chromosome is lost or occluded in cell maturation. In simultaneous double hybridisation with both probes, HPV and Y sequences were demonstrable within the same nucleus in penile warts. This technique permits the simultaneous differential detection of two nuclei acid sequences in interphase nuclei and will have application in analysis of putative dual HPV infections and in determining the intranuclear spatial relations between nucleic acids in interphase nuclei. PMID- 2544634 TI - Basic and clinical pharmacology of amiodarone: relationship of antiarrhythmic effects, dose and drug concentrations to intracellular inclusion bodies. AB - Amiodarone is a unique class III antiarrhythmic drug with several unusual pharmacokinetic, pharmacodynamic, and toxicological actions which are quite distinct from those of the standard antiarrhythmic drugs. Extensive animal and clinical studies have demonstrated that amiodarone and its major metabolite, desethylamiodarone, both produce a marked increase in the duration of transmembrane action potential, which may be related to their antiarrhythmic as well as clinical electrophysiological activity. Unlike most other cardiovascular drugs, it has been recognized for more than 20 years that optimal antiarrhythmic effects may take several days to weeks after onset of oral therapy. Amiodarone is highly lipid soluble and exhibits at least three separate compartments of drug distribution, with a long elimination half-life of 14-120 days after chronic therapy. The pharmacokinetic profile of desethylamiodarone is qualitatively similar to that of amiodarone, but its elimination half-life is even longer and its tissue distribution may be slightly different. Although there may not be any correlation between serum drug levels and clinical toxicity of amiodarone during long-term therapy, recent animal as well as clinical data suggest that multilamellar intracellular inclusions can be dissociated from cell death or clinical toxicity. Thus, it is possible that amiodarone toxicity can be minimized with low doses or low serum drug concentrations. The metabolite(s) of amiodarone may play a major role in its pharmacological and toxicological actions. PMID- 2544635 TI - Effects of two inotropic drugs, dopamine and dobutamine, on pulmonary gas exchange in artificially ventilated patients. AB - The inotropic agents, dopamine (DP) and dobutamine (DB), both decrease PaO2, probably by a redistribution of the VA/Q ratio. The aim of this study was to assess the effect of both drugs on the VA/Q ratio, using the multiple inert gas elimination method. Ten artificially ventilated patients (eight males), aged 45 74 years were investigated. Blood gases, cardiac output and concentrations of inert gases were measured before and 30 min after infusion of DB or DP. DP and DB were administered alternatively at a rate of 5 micrograms.k-1 min-1. The decrease in PaO2 was significantly greater with DP (12 +/- 9 torr) than with DB (7 +/- 9 torr) (P less than 0.01). Both drugs similarly increased cardiac output: +2.61.min-1 +/- 1.4 for DP and 2.21.min-1 +/- 1.5 for DB. Both DP and DB significantly (P less than 0.01) increased the perfusion of alveoli with VA/Q = 0 (+4 +/- 7% for DP and +3 +/- 7% for DB) and 0 less than VA/Q less than 0.1 (+11 +/- 8.5% for DP and +5.5 +/- 10.5% for DB) (no significant difference between the drugs). When shunt and "shunt-like" effect are considered together, there was a significantly greater increase in the amount of blood going to alveoli with a low VA/Q ratio with DP compared to DB. Both drugs decreased the perfusion of alveoli with 0.1 less than VA/Q less than 10, but the decrease was significantly less for DB than for DP (-15 +/- 6.5% for DP and -8.5 +/- 7% for DB, p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544636 TI - The influences of dexamethasone levels on the predictive value of the DST for unipolar major depression and the relationships between post-dexamethasone cortisol and ACTH levels. AB - To investigate the relationships between dexamethasone (DEX) and post-DEX cortisol and adrenocorticotropic hormone (ACTH) levels, the authors measured DEX at 8.00 a.m. and post-DEX cortisol and ACTH levels at 8.00 a.m. and 4.00 p.m. in 72 depressed patients categorized according to DSM-III. Cortisol non-suppressors exhibited significantly (P = 0.0006) decreased levels of DEX compared to suppressors. DEX levels at 8.00 a.m. explained 21.1% of the variance in the post DEX cortisol values at 8.00 a.m. and 34.5% of those at 4.00 p.m. DEX levels were not significantly different among minor depressives (300.40, 309.00), major depressives without melancholia (296.X2) or with melancholia and/or psychotic features (296.X3, 296.X4). In the latter the post-DEX cortisol was significantly increased compared to all other depressives and these differences remained significant even after adjusting for the variations in DEX (by means of regression analysis). Also the diagnostic performance of the post-DEX cortisol values for major depression with associated features versus minor depression was not substantially affected when the DEX levels were accounted for. ACTH levels after DEX were shown to correlate significantly (P less than 0.05) and negatively with DEX. Although post-DEX ACTH levels did not differ among the DSM-III diagnostic categories, cortisol non-suppressors averaged significantly (P = 0.0004) higher ACTH levels than suppressors. PMID- 2544637 TI - Plasma N-POMC, ACTH and cortisol following hCRH administration in major depression and dysthymia. AB - The concentrations of plasma N-terminal pro-opiomelanocortin, adrenocorticotropic hormone and cortisol in response to a 14:30 h human corticotrophin releasing hormone (hCRH) stimulation test (1 microgram/kg) were measured in control, major depression and dysthymic subjects. The increases in all three hormones were similar in the depressed groups when compared to the control values. The elevation in cortisol after hCRH was significantly greater in major depression when compared to the dysthymic subjects. The significance of these findings is discussed. PMID- 2544639 TI - The release of neutrophil chemoattractant activity by bronchoalveolar macrophages from adult and senescent mice. AB - To assess the effects of advanced age on the release of neutrophil chemoattractant activity by resident bronchoalveolar macrophages (BAM), cells from three strains of pathogen- and disease-free mice were secured by lung lavage and stimulated in vitro with unopsonized zymosan or the calcium ionophore, A23187. Chemoattractant release by BAM from adult (5-8 mos) and senescent (19-26 mos) C57BL/6 and DBA/2 mice in response to both stimulants was comparable; however, the generation of chemoattractant activity by BAM from senescent (18-20 mos) BALB/c mice was greater than that observed with cells from younger (4-6 mos) animals with both zymosan and A23187. In the presence of 50 microM piriprost potassium, a 5-lipoxygenase inhibitor, the release of chemoattractant activity and leukotriene B4 (LTB4) in response to zymosan and A23187 by BAM from both groups of C57BL/6 mice was significantly impaired. With BAM from BALB/c mice, 100 microM piriprost potassium was required to produce changes in A23187-stimulated chemoattractant and LTB4 release; of note, the generation of LTB4 in response to A23187 by BAM from aged BALB/c mice was significantly greater than that observed with cells from the younger animals under all conditions studied. Finally, with BAM from DBA/2 mice, 50 microM piriprost potassium significantly reduced chemoattractant activity in both groups of animals, but the lipoxygenase inhibitor had no effect on LTB4 production. Thus, although these studies revealed substantial age and strain-related differences in the release of neutrophil chemoattractant activity by murine BAM, they did not demonstrate deficiencies that might enhance the susceptibility of the senescent host to infection of the lower respiratory tract. PMID- 2544638 TI - Platelet serotonin uptake and 3H-imipramine binding in panic disorder. AB - Platelet serotonin uptake and 3H-imipramine binding were measured in eight patients with panic disorders and nine controls. The Vmax of serotonin uptake was significantly elevated in patients compared to controls (77 +/- 14 vs. 50 +/- 4 pmol/10(8) platelets/min; P less than 0.05) while Km values were not different (1.46 +/- 0.41 vs. 1.24 +/- 0.20 microM). 3H-Imipramine binding to ruptured platelet membranes was not significantly different between patients and controls for either Bmax (395 +/- 71 vs. 412 +/- 107 fmol/mg protein) or Kd (0.90 +/- 0.18 vs. 1.09 +/- 0.30 nM). The implications for a serotonergic dysfunction in panic disorders are discussed. PMID- 2544640 TI - Bupivacaine: a safe local anesthetic for wrist blocks. AB - Seventy-one patients having minor hand surgical procedures under wrist block anesthesia were studied, with the goal of identifying a possible neurotoxic potential of bupivacaine when used according to standard clinical practice. This drug was compared with lidocaine at equipotent analgesic concentrations (bupivacaine: 5 mg/ml; lidocaine: 20 mg/ml) by use of a double-blind randomized protocol. At postoperative follow-up, four (5.6%) patients had new neurologic symptoms. Mechanical factors, e.g., nerve manipulation during surgery, or nerve trauma during performance of the block, were identified in three patients. In the remaining patient, anesthesia was induced with lidocaine, and no cause could be identified. It was concluded that bupivacaine, when used in clinical concentrations, is not associated with an increased incidence of neural complications. PMID- 2544641 TI - Sweat gland carcinoma of the hand (malignant acrospiroma). AB - A case of malignant acrospiroma of the hand is reported. This rare sweat gland carcinoma recurs locally in 50% of patients, with distant metastases occurring in 60% of patients. The prognosis for 5-year disease-free survival is less than 30%. Although the histopathologic findings of tumor-free margins and negative regional lymph node dissection indicate a good prognosis in most carcinomas, they do not guarantee a good prognosis in malignant acrospiroma because of the aggressive characteristics of this tumor. PMID- 2544642 TI - Radial nerve laceration and retraction associated with a supracondylar fracture of the humerus. AB - Supracondylar fracture of the humerus can be associated with varying degrees of radial nerve injury. Complete laceration of the nerve is rare. We report a case of radial nerve laceration after a supracondylar fracture in which the proximal segment of the nerve retracted into the axilla. Successful treatment resulted from long segmental sural nerve grafting. PMID- 2544643 TI - Infection control: new problem organisms for infection control. PMID- 2544644 TI - Differential expression of functional adrenocorticotropic hormone receptors by subpopulations of lymphocytes. AB - In an effort to investigate the presence of adrenocorticotropic hormone (ACTH) receptors on rat lymphocytes, cells were separated by a panning procedure into T and B cell populations. By using the radiolabeled ACTH agonist, (125I-Tyr23) phenylalanine2-norleucine4-ACTH1-24, substantial numbers of ACTH binding sites were detected on T and B lymphocytes, but not on thymocytes. Scatchard analysis revealed two types of binding sites on each cell population, one with Kd1 = 0.088 +/- 0.025 nM and one with Kd2 = 4.2 +/- 0.6 nM; however, the absolute number of binding sites per cell was different. B lymphocytes expressed approximately three times the number of Kd1 binding sites per cell when compared with T lymphocytes. However, ACTH receptor expression by these cell populations was not static as suggested by the ability to induce receptor expression via mitogens. B or T cells and thymocytes stimulated with the mitogens LPS or Con A, respectively, substantially increased their number of Kd1 binding sites per cell (approximately three-fold). Even more dramatic increases in Kd1 receptor expression (approximately 100-fold) were observed when comparing "normal" and stimulated thymocytes. To demonstrate that these ACTH binding sites were in fact functional, cAMP levels were measured in lymphocytes 10 min after exposure to varying concentrations of ACTH. Dose-dependent increases in cAMP levels were observed, with significant stimulation occurring with as little as 0.1 nM ACTH added. Taken together, these studies demonstrate the presence of functional ACTH receptors on normal, rat T and B lymphocytes. PMID- 2544645 TI - Viral antigen stimulation of the production of human monokines capable of regulating HIV1 expression. AB - We have previously described model systems for cytokine-induced regulation of chronically HIV-infected promonocyte and T cell clones. Using these systems, we have shown that monokines contained in supernatants from LPS-stimulated human monocyte/macrophages (MO) up-regulate HIV expression, reflected by an increase in reverse transcriptase activity, viral RNA levels, and expressed viral proteins. Current studies were designed to determine whether viral Ag can interact with MO and secondarily affect HIV1 expression by stimulating monokine production. We found that certain herpes-group viruses, including CMV and EBV, augment HIV1 expression by inducing monokine production, whereas others, such as HSV1, HSV2, varicella-zoster virus, and human herpes virus 6 were unable to function in this capacity. The HSV1 and HSV2 Ag which failed to stimulate monokine production did not interfere with MO stimulation by CMV Ag, suggesting that failure to induce HIV expression was not attributable to MO suppression. When nonherpes group viruses were tested, we found that human adenovirus, hepatitis B virus, and vaccinia virus all failed to stimulate the production of monokines capable of activating HIV in the chronically infected cell lines. In contrast, HIV1 can augment its own expression by inducing the secretion of monokines which up regulate HIV expression in the infected cells. The viral Ag-induced MO supernatants capable of up-regulating HIV expression did so in a dose-dependent manner, whereas viral Ag alone produced no significant change. Monokine production mediated by viral Ag was not attributable to contaminating endotoxin. These studies provide a model to determine whether other opportunistic infections may induce the expression of HIV by indirect mechanisms, such as the stimulation of cytokine production. PMID- 2544646 TI - Cholera toxin as a mucosal adjuvant. Glutaraldehyde treatment dissociates adjuvanticity from toxicity. AB - Cholera toxin (CT), either mixed with or conjugated to unrelated protein Ag, is known to enhance the intestinal IgA response of rodents toward the unrelated Ag. Although relatively low doses of CT exert this gut mucosal adjuvant effect, the inherent toxicity of CT is a hindrance to its use in humans. Our report demonstrates that CT treated with 20 mM glutaraldehyde retains adjuvant properties but exhibits more than 1000-fold lower toxicity than untreated toxin. Glutaraldehyde was also used in a one-stage conjugation procedure to couple CT covalently to Sendai virus. Again, toxicity was reduced more than 1000-fold. This drop in toxicity is consistent with an observed 100-fold loss in binding capacity of the CT B subunit and a 20- to 50-fold reduction in adenylate cyclase activation by the CT A subunit. Oral administration of this virus-toxoid conjugate resulted in increased gut antiviral IgA titers compared with oral administration of either virus alone or of virus mixed with glutaraldehyde treated toxin. This marked decrease in toxicity may afford a practical approach for the use of CT as a mucosal adjuvant. PMID- 2544647 TI - Neutrophil cathepsin G increases calcium flux and inositol polyphosphate production in cultured endothelial cells. AB - Exposure of endothelial cells (ENDO) to human neutrophil cathepsin G (CG) increases albumin flux across the endothelial monolayer. Since calcium influences cell shape and barrier function of ENDO monolayers, the current study was designed to determine if CG acted through alterations in Ca2+ homeostasis in ENDO. The role of Ca2+ in the increased permeability of ENDO monolayers to albumin after exposure to CG was studied by using ENDO monolayers cultured on polycarbonate filters. Exposure of ENDO monolayers to CG in the presence of the Ca2+-antagonist lanthanum partially prevented the increase in albumin flux, but exposure in the presence of agents that block voltage-regulated calcium channels did not block the increase in albumin flux. To monitor the effect of CG on Ca2+ flux, ENDO were labeled with 45Ca2+ and changes in Ca2+ flux were monitored by the release of 45Ca2+. From 1 to 15 minutes after exposure of ENDO to CG, there was increased release of 45Ca2+ compared with control cells. Calcium channel blocking agents did not inhibit the increased release of 45Ca2+, but lanthanum partially blocked the increase. The increased release of Ca2+ appeared to be due, at least in part, to activation of phospholipase C because there was an increase both in inositol polyphosphate species and in diglycerides after incubation of ENDO with CG. These studies support the hypothesis that CG increases the flux of calcium in ENDO, that this increase in Ca2+ flux may result from activation of phospholipase C, and that this system may be involved in the decreased barrier properties of the ENDO after CG exposure. PMID- 2544648 TI - Effects of treatment with IL-2 receptor specific monoclonal antibody in mice. Inhibition of cytotoxic T cell responses but not of T help. AB - Contribution of IL-2R-bearing activated lymphocytes to antiviral host defense was investigated in C57BL/6 mice by treatment in vivo with IL-2R-specific mAb PC61. When treated on days 0 and 1 with respect to infection with either vaccinia virus, lymphocytic choriomeningitis (LCM) virus (LCMV) or vesicular stomatitis virus, 6-day immune mice had low numbers of CD8+ T cells that were reduced to about 10% of the values found for infected but otherwise untreated controls. In contrast, the number of CD4+ T cells was within normal ranges. Correspondingly, induction of strictly T help-dependent antiviral neutralizing IgG antibody titers remained unaffected by the mAb treatment, whereas generation of antiviral cytotoxic T cell activity was abrogated. Anti-IL-2R treatment of thymectomized mice 14 and 15 days after infection prevented generation of secondary antiviral cytotoxic T cells in restimulation cultures in vitro initiated 24 days later. Treatment with IL-2R-specific mAb was comparable to treatment with CD8-specific mAb in preventing mice to eliminate virus. Because of the involvement of antiviral cytotoxic T cells in disease manifestations, treatment with IL-2R specific mAb protected mice from lethal LCM after intracerebral infection with LCMV and inhibited the footpad swelling reaction caused by local infection with the same virus. PMID- 2544649 TI - Feline cytotoxic large granular lymphocytes induced by recombinant human IL-2. AB - Large granular lymphocytes (LGL) have been characterized phenotypically and functionally as cytotoxic T lymphocytes, NK cells or lymphokine-activated killer cells. The most prominent morphologic feature of LGL is large cytoplasmic granules that are thought to contain the molecules responsible for cell lysis. In this study, we describe the morphologic and functional characteristics of IL-2 dependent cytotoxic lymphocytes derived from feline PBL. Stimulation of feline PBL with Con A followed by culturing in 50 U of gibbon monkey IL-2 human rIL-2 induced long term lymphocyte cultures. These lymphocytes are cytotoxic for the feline leukemia virus-induced T cell lymphoma (FL74), in a 4-h 51Cr release assay. All cell lines are either constitutively cytotoxic for FL74 cells, or cytotoxic in a lectin-dependent cell cytotoxic assay, the latter being a characteristic of low passage cultures. In contrast, no cell lines express self lysis or lysis for other lines. [3H]TdR uptake showed that 1 U of human rIL-2 produces a 50% maximal proliferative response by feline lymphocytes suggesting a high degree of homology between the ligand binding sites of feline and human IL 2R. Feline cytotoxic lymphocytes possess abundant cytoplasm containing large azurophilic granules characteristic of LGL. These granules are bound by a bilipid membrane and contain numerous smaller membrane-bound vesicles 50 to 60 nm in diameter. A model is proposed, whereby subsequent to binding of LGL to target cell the large granules fuse to the LGL plasma membrane and release the small vesicles into the binding pocket. The vesicles then transport the lytic molecules directly and selectively to the target cell membrane. PMID- 2544650 TI - [Increase in the generation of superoxide radicals and in the inhibitory effect on Yoshida sarcoma of anthracycline antitumor agents by ultrasound]. AB - The antitumor effects of ultrasonic (US) irradiation in combination with the administration of an anthracycline drug, such as adriamycin and THP-adriamycin, were investigated from a viewpoint of the generation of superoxide radicals (SOR). In the in vitro experiments, the generation of SOR by US irradiation was measured by the amount of cytochrome c reduced. The addition of the drug stimulated the generation of SOR by US irradiation. In the in vivo experiments, Donryu rats inoculated subcutaneously by Yoshida sarcoma were treated with US irradiation in combination with the drug. During US irradiation, the temperature of the rat tissue irradiated was kept below 37 degrees C to avoid thermal effects. To know the optimum timing of US irradiation after the administration of the drug, the drug concentrations in the tumor and blood were determined and the time course of drug concentrations was analyzed pharmacokinetically. The effects of drugs and/or US irradiation showed antitumor activity judged by the growth of the tumor size or the survival time of rats. The combination treatments of US irradiation with the drug marked additional or synergistic effects on Yoshida sarcoma. Considering the relationship between the antitumor effect in vivo and the generation of SOR in vitro, the increase of anti-tumor effect of US irradiation by the anthracycline drug may be caused by the stimulation of the generation of SOR. PMID- 2544651 TI - [Clinical studies of intermittent hepatic arterial occlusion with infusion chemotherapy for unresectable hepatocellular carcinoma associated with arterioportal or arteriovenous shunts]. AB - Intermittent hepatic artery occlusion combined with infusion chemotherapy is a newly devised methodology. A double lumen balloon catheter was surgically inserted into the hepatic artery. Through the catheter, fluorouracil (5-Fu) was continuously infused and mitomycin (MMC) or adriamycin (ADM) was injected in one shot at the time of blood flow occlusion. This new methodology was performed in 19 patients who had unresectable hepatocellular carcinoma. Although five of 19 cases had arterioportal (A-P) or arteriovenous (A-V) shunts, four of them also responded well and objective anti-tumor effects resulted. In addition, complications associated with A-P or A-V shunts such as bleeding from the digestive tract due to portal hypertension were well managed by this new methodology. PMID- 2544652 TI - Decreased erythrocyte phosphoribosylpyrophosphate synthetase activity and impaired formation in thalassemia minor: a mechanism for decreased adenine nucleotide content. AB - Adenosine triphosphate (ATP) and adenosine diphosphate levels are decreased in erythrocytes from individuals with beta-thalassemia minor. Because 5 phosphoribosyl-1-pyrophosphate (PRPP) is an essential precurosr of adenine nucleotides, we tested the hypothesis that impaired PRPP synthesis is a mechanism for the decreased adenine nucleotide content. Erythrocyte PRPP synthetase activity was significantly decreased, and the Michaelis-Menten constant (Km) for ribose-5-phosphate (R5P) was significantly increased in individuals with alpha thalassemia minor and those with beta-thalassemia minor. Intact erythrocytes from individuals with alpha-thalassemia and those with beta-thalassemia minor also had an impaired rate of PRPP formation. Both the decrease in PRPP synthetase activity and the impaired PRPP formation were also found in erythrocytes with microcytosis resulting from iron deficiency, indicating that these phenomena may not be specific to thalassemia minor. In all individuals examined, the rate of PRPP formation correlated with ATP content, suggesting that either (1) PRPP synthetase activity is a determinant of ATP content or (2) ATP content is a determinant of PRPP synthetase activity. The depletion of ATP from normal erythrocytes did not affect PRPP synthetase activity, suggesting that ATP content is not a determinant of PRPP synthetase activity. However, a decrease in PRPP synthetase activity did cause an impairment in the rate of adenine nucleotide synthesis, suggesting that PRPP synthetase activity is a determinant of ATP content. Taken together, our results suggest that the decrease in PRPP synthetase activity and the resulting impairment in the rate of PRPP formation are mechanisms for the decreased adenine nucleotide content in thalassemic erythrocytes. PMID- 2544653 TI - Neutrophil oxidative metabolism after exposure to bacterial phospholipase C. AB - The effects of phospholipase C (PLC) from Clostridium perfringens and Bacillus cereus on bovine neutrophil oxidative metabolism were studied by measuring superoxide production and oxygen consumption in response to PLC alone or in combination with other stimuli. PLC from both species elicited superoxide production and enhanced the response to stimulation by NaF when the two stimuli were given simultaneously. However, oxygen consumption in response to latex beads was markedly inhibited by pretreatment of cells with B. cereus PLC. The results suggest that some bacterial PLC exoenzymes may initially activate neutrophils but leave the cells relatively unresponsive to subsequent stimuli. PMID- 2544654 TI - Urinary oestriol-16-glucuronide determined by "on-line" liquid chromatography. AB - A fully automated method for the specific assessment of oestriol-16-glucuronide in urine is described. "On-line" sample preparation, including enrichment, pre purification, focusing and injection, is combined with automated high-performance liquid chromatographic separation and fluorimetric quantification. Losses of oestriol-16-glucuronide throughout the total procedure are negligible. Thus, external calibration is feasible for quantification. Coefficients of variation are 4.44% (n = 12) for intra- and 7.85% (n = 9) for interassay variability. Assay sensitivity is 430 nmol/l. The excretion rates of oestriol-16-glucuronide relative to creatinine were estimated in 85 pregnancy urines. These oestriol-16 glucuronide estimates correlated well with those of total urinary oestriol, determined by high-performance liquid chromatography after acid hydrolysis (r = 0.957). The reference ranges of oestriol-16-glucuronide for the different weeks of gestation were evaluated. Unlike the determination of total oestriol, the present method does not need an hydrolysis step. The method is therefore well suited for the biochemical monitoring of fetal well-being under emergency conditions. PMID- 2544657 TI - Biology and pathogenesis of lentiviruses. PMID- 2544656 TI - Ionic currents in dispersed chemoreceptor cells of the mammalian carotid body. AB - Ionic currents of enzymatically dispersed type I and type II cells of the carotid body have been studied using the whole cell variant of the patch-clamp technique. Type II cells only have a tiny, slowly activating outward potassium current. By contrast, in every type I chemoreceptor cell studied we found (a) sodium, (b) calcium, and (c) potassium currents. (a) The sodium current has a fast activation time course and an activation threshold at approximately -40 mV. At all voltages inactivation follows a single exponential time course. The time constant of inactivation is 0.67 ms at 0 mV. Half steady state inactivation occurs at a membrane potential of approximately -50 mV. (b) The calcium current is almost totally abolished when most of the external calcium is replaced by magnesium. The activation threshold of this current is at approximately -40 mV and at 0 mV it reaches a peak amplitude in 6-8 ms. The calcium current inactivates very slowly and only decreases to 27% of the maximal value at the end of 300-ms pulses to 40 mV. The calcium current was about two times larger when barium ions were used as charge carriers instead of calcium ions. Barium ions also shifted 15-20 mV toward negative voltages the conductance vs. voltage curve. Deactivation kinetics of the calcium current follows a biphasic time course well fitted by the sum of two exponentials. At -80 mV the slow component has a time constant of 1.3 +/- 0.4 ms whereas the fast component, with an amplitude about 20 times larger than the slow component, has a time constant of 0.16 +/- 0.03 ms. These results suggest that type I cells have predominantly fast deactivating calcium channels. The slow component of the tails may represent the activity of a small population of slowly deactivating calcium channels, although other possibilities are considered. (c) Potassium current seems to be mainly due to the activity of voltage-dependent potassium channels, but a small percentage of calcium-activated channels may also exist. This current activates slowly, reaches a peak amplitude in 5-10 ms, and thereafter slowly inactivates. Inactivation is almost complete in 250-300 ms. The potassium current is reversibly blocked by tetraethylammonium. Under current clamp conditions type I cells can spontaneously fire large action potentials. These results indicate that type I cells are excitable and have a variety of ionic conductances. We suggest a possible participation of these conductances in chemoreception. PMID- 2544655 TI - Stoichiometry and voltage dependence of the sodium pump in voltage-clamped, internally dialyzed squid giant axon. AB - The stoichiometry and voltage dependence of the Na/K pump were studied in internally dialyzed, voltage-clamped squid giant axons by simultaneously measuring, at various membrane potentials, the changes in Na efflux (delta phi Na) and holding current (delta I) induced by dihydrodigitoxigenin (H2DTG). H2DTG stops the Na/K pump without directly affecting other current pathways: (a) it causes no delta I when the pump lacks Na, K, Mg, or ATP, and (b) ouabain causes no delta I or delta phi Na in the presence of saturating H2DTG. External K (Ko) activates Na efflux with Michaelis-Menten kinetics (Km = 0.45 +/- 0.06 mM [SEM]) in Na-free seawater (SW), but with sigmoid kinetics in approximately 400 mM Na SW (Hill coefficient = 1.53 +/- 0.08, K1/2 = 3.92 +/- 0.29 mM). H2DTG inhibits less strongly (Ki = 6.1 +/- 0.3 microM) in 1 or 10 mM K Na-free SW than in 10 mM K, 390 mM Na SW (1.8 +/- 0.2 microM). Dialysis with 5 mM each ATP, phosphoenolpyruvate, and phosphoarginine reduced Na/Na exchange to at most 2% of the H2DTG-sensitive Na efflux. H2DTG sensitive but nonpump current caused by periaxonal K accumulation upon stopping the pump, was minimized by the K channel blockers 3,4-diaminopyridine (1 mM), tetraethylammonium (approximately 200 mM), and phenylpropyltriethylammonium (20-25 mM) whose adequacy was tested by varying [K]o (0-10 mM) with H2DTG present. Two ancillary clamp circuits suppressed stray current from the axon ends. Current and flux measured from the center pool derive from the same membrane area since, over the voltage range -60 to +20 mV, tetrodotoxin-sensitive current and Na efflux into Na-free SW, under K-free conditions, were equal. The stoichiometry and voltage dependence of pump Na/K exchange were examined at near-saturating [ATP], [K]o and [Na]i in both Na-free and 390 mM Na SW. The H2DTG-sensitive F delta phi Na/delta I ratio (F is Faraday's constant) of paired measurements corrected for membrane area match, was 2.86 +/- 0.09 (n = 8) at 0 mV and 3.05 +/- 0.13 (n = 6) at -60 to -90 mV in Na free SW, and 2.72 +/- 0.09 (n = 7) at 0 mV and 2.91 +/- 0.21 (n = 4) at -60 mV in 390 mM Na SW. Its overall mean value was 2.87 +/- 0.07 (n = 25), which was not significantly different from the 3.0 expected of a 3 Na/2 K pump.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2544658 TI - Asymptomatic infection of the central nervous system by the macaque immunosuppressive type D retrovirus, SRV-1. AB - The aetiological agent of spontaneously occurring simian acquired immune deficiency syndrome (SAIDS) in rhesus monkeys (Macaca mulatta) at the California Primate Research Center is a type D retrovirus designated SAIDS retrovirus serotype 1 (SRV-1). SRV-1 DNA and RNA have previously been detected in the brains of rhesus monkeys with SAIDS in the absence of viral antigen or neuropathological lesions. In this study we further define the relationship between SRV-1 and the central nervous system (CNS) in rhesus monkeys by examining the CNS for infectious SRV-1, viral antigen and anti-SRV-1 antibodies. In addition, cerebrospinal fluid (CSF) was assayed for alterations in IgG and albumin levels, IgG/albumin ratios and cell count in comparison to uninfected control animals. No differences in CSF parameters were detected between infected and uninfected animals except for the presence of infectious SRV-1 which was isolated from the CSF from 13 out of 19 (68%) viraemic rhesus monkeys. The probable source of this virus was the choroid plexus, where approximately 1 in 1000 surface epithelial cells were found to contain viral antigen by immunohistochemistry. Antibodies against SRV-1 were not detected in the CSF even when present in the serum. Neither infectious virus nor viral antigen were found in the brain parenchyma of any animal examined. Thus infection of the CNS by SRV-1 appears to be subclinical without an intrathecal immune response. This may be related to the apparent restriction of productive infection in the CNS to cells of the choroid plexus. PMID- 2544660 TI - Completion of the sequence of bluetongue virus serotype 10 by the characterization of a structural protein, VP6, and a non-structural protein, NS2. AB - The sequence of cDNA clones representing the entire genome of bluetongue virus serotype 10 (BTV-10) has been completed by the analysis of data obtained for the S8 and S9 segments. Each DNA clone has been sequenced completely and the deduced amino acid sequences have been analysed. The sequences of the S8 and S9 gene products as well as another two previously published small gene products (S7 and S10) have been compared with the corresponding size gene products of reovirus type 1. The data do not indicate a relationship between the small proteins of these two viruses except some distant homologies between the BTV VP7 protein and the sigma 1 protein of reovirus. The characteristics of all the BTV-10 genome segments, the deduced primary gene products and their possible functions are summarized. PMID- 2544661 TI - Genotypic selection following coinfection of cultured cells with subgroup 1 and subgroup 2 human rotaviruses. AB - The purpose of this study was to determine why identifiable reassortants between subgroup 1 and subgroup 2 rotaviruses have been so rarely isolated from human specimens. Cultured cells were coinfected with pairs of subgroup 1 and 2 human rotaviruses and passaged multiple times to simulate natural reassortant formation and selection in vivo. After coinfection of MA-104 cells with subgroup 1 (DS-1) and subgroup 2 (either Wa or P) strains, approximately 14% of the plaque-picked progeny were shown to be reassortants. During multiple passages of these coinfected cultures, however, complete (Wa virus coinfection) or nearly complete (P virus coinfection) loss of detectable DS-1 segments from progeny was observed. Thus, when all segments of the subgroup 2 viruses were present in coinfected cultures, these segments dominated in the selected progeny. Coinfection with subgroup 1-subgroup 2 rotavirus reassortants and the DS-1 strain followed by multiple passages, however, resulted in complete loss of some segments from the subgroup 2 strains originally present in the reassortants. Therefore, segments from the parental subgroup 2 viruses appeared to be selected in toto during multiple passages because they were dominant as a group, not because individual segments of these viruses were consistently favoured over their subgroup 1 virus counterparts. PMID- 2544659 TI - Bluetongue virus infection of bovine monocytes. AB - Cultures of adherent and non-adherent bovine blood mononuclear cells were infected with bluetongue virus (BTV) serotype 10. Production of BTV proteins in mononuclear cell cultures was detected by immune precipitation of viral proteins from [35S]methionine-labelled extracts of these cells, by immunofluorescence staining of cells using monoclonal antibodies (MAbs) to BTV proteins VP7 and NS2, and by flow cytometry with MAbs to VP2, VP7, NS1 and NS2. BTV-infected cells were most numerous in cultures of adherent mononuclear cells; infected cells were initially identified as monocytes on the basis of their morphology, and size and scatter characteristics as determined by analysis with a fluorescence-activated cell sorter (FACS). The majority of adherent mononuclear cells with these scatter characteristics were confirmed to be monocytes by FACS analysis with a MAb specific for bovine monocytes. Identification of BTV-infected adherent mononuclear cells as monocytes was further established by double immunofluorescent labelling, as infected adherent cells reacted with the MAb specific for bovine monocytes, and with another MAb specific for class II antigen. Infection of adherent mononuclear cells was also confirmed by transmission electron microscopy, as BTV virions and tubules were present in lysates of cultures of BTV-infected adherent mononuclear cells and within the cytoplasm of adherent cells. In contrast, BTV proteins were detected in few cells identified as lymphocytes on the basis of their scatter characteristics, and mean fluorescence of such cells was considerably less than that of BTV-infected monocytes. Viraemia persisted until 35 days after inoculation of a colostrum deprived calf inoculated with BTV. Virus was isolated from blood mononuclear cells at 1 week after infection of the calf, but not thereafter. BTV infection of blood mononuclear cells was demonstrated until 9 days after inoculation by indirect immunofluorescence staining of mononuclear cells. In contrast, virus was consistently isolated from erythrocyte-enriched preparations throughout viraemia in titres comparable to those in whole blood. These results indicate that although bovine monocytes are readily infected in vitro with this strain of BTV serotype 10, infection of blood monocytes is unlikely to be responsible for the prolonged viraemia that consistently occurs in BTV-infected cattle. PMID- 2544662 TI - Several rat cell lines share a common defect in their inability to internalize murine coronaviruses efficiently. AB - Infection of rat cells, Schwannoma RN2, hepatoma HTC or myoblast L6, with the murine coronavirus JHM strain results in a persistent infection characterized by the release of virus over an extended period of time with a limited cytopathology. Several stages of the viral replication cycle have been examined in these cells in comparison to those in mouse L2 cells, which are totally permissive to JHM infection. Although the rat cells bound as much virus as the mouse cells. Their ability to internalize it was 40-fold less efficient than the mouse cells. This lower internalization efficiency was not enhanced by pH shock of infected cells, but was by treatment with polyethylene glycol. In all cell types there appeared to be no major differences in the ability of the internalized virus to replicate the viral RNA as determined by slot-blot analysis with a radiolabelled viral cDNA. A similar genetic mechanism appears to be operative in the lines because somatic cell hybrids formed between these lines in various combinations were also deficient in the ability to internalize bound virus. Taken together these results imply that rat cell lines in general share a common deficiency in their inability to internalize murine coronaviruses efficiently. This low efficiency in viral internalization may explain in part the ability of these lines to sustain persistent infections. PMID- 2544663 TI - Epstein-Barr virus latent gene expression during the initiation of B cell immortalization. AB - Epstein-Barr virus (EBV) has the capacity to immortalize a subpopulation of resting B lymphocytes. Lymphoblastoid cell lines (LCL) established in this way carry the latent EBV genome as multiple copies of an extrachromosomal episome. Viral gene expression in LCLs is highly restricted; products identified correspond to a membrane protein (latent membrane protein; LMP), a nuclear antigen complex (Epstein-Barr nuclear antigens; EBNAs 1 to 6), two small RNA species (EBERs 1 and 2) and RNA species thought to encode a second membrane associated polypeptide designated terminal protein (TP). Here we have investigated the temporal sequence of expression of the characterized 'latent' proteins during the initiation of immortalization when resting B cells are stimulated to enter and traverse the cell cycle. The analysis has been carried out on prolymphocytic leukaemia cells infected in vitro with either the immortalizing B95-8 strain of virus or the non-immortalizing P3HR1 strain. The results reveal that following B95-8 infection, a sequence of EBV expression is initiated within approximately 8 h with the synthesis of detectable levels of EBNA 2 shortly followed by EBNAs 1, 3, 4, 5 and 6. There is then a delay of approximately 40 h until the expression of LMP completes the latent pattern of proteins found in LCLs. P3HR1 infection, however, produces only transient expression of some EBNA species in a small percentage of cells after approximately 48 h. These observations suggest the failure of P3HR1 virus to immortalize may not be due solely to the absence of EBNA 2 expression and that cellular and/or virus-mediated events occur after EBNA synthesis which then facilitate efficient LMP expression and immortalization. PMID- 2544664 TI - Effects of L3T4+ lymphocyte depletion on acute murine cytomegalovirus infection. AB - We examined the role of T lymphocytes bearing the L3T4 phenotype in acute murine cytomegalovirus (MCMV) infection. In vivo administration of rat IgG2b monoclonal antibody (MAb) GK 1.5 was used to deplete mice of L3T4+ lymphocytes during acute MCMV infection. Unlike the saline-treated controls that resolved their infections, mice receiving the MAb developed persistent and high levels of virus in the salivary gland, lung and spleen. The production of antibody to MCMV was delayed and the titres achieved were markedly less than in the controls. Despite the higher levels of virus replication, there was no increase in mortality seen in animals treated with the MAb. Following intraperitoneal challenge with MCMV, depletion of L3T4+ lymphocytes was protective, increasing the dose of MCMV required to produce death. These data indicate that T lymphocytes of the L3T4 phenotype influence the degree of MCMV replication during acute infection and may contribute to mortality following intraperitoneal virus challenge. PMID- 2544665 TI - Effects of acyclovir on herpes simplex virus type 1 infection in mice treated with 12-O-tetradecanoylphorbol 13-acetate. AB - The purpose of this study was to determine whether infectious herpes simplex virus type 1 (HSV-1) has tumorigenic properties and, if so, whether inhibition of the cytolytic replicative cycle of the virus after infection enhances tumour development. Eighty mice were subjected to repeated inoculation of HSV-1 on their upper lips after scarification, and systemic administration of acyclovir (ACV). 12-O-tetradecanoylphorbol 13-acetate (TPA) was used as the tumour promoter. The tumour incidence was compared to control groups each of 40 mice that were either not treated with ACV, not treated with TPA, not infected with HSV or only scarified. In the virus-infected group treated with ACV and TPA, 25% of the animals developed tumours. In the HSV-infected group treated with TPA only, 25% of the animals also developed tumours. The uninfected animals which were not treated with TPA developed tumours to a significantly lesser degree. In conclusion, the combined effects of HSV-1 and TPA, with or without ACV treatment, resulted in a significant increase in the number of tumours in comparison to the control groups. PMID- 2544666 TI - Nucleotide sequence and characterization of the Marek's disease virus homologue of glycoprotein B of herpes simplex virus. AB - The Marek's disease virus (MDV) homologue of the herpes simplex virus (HSV) gene encoding glycoprotein B (gB) has been identified within BamHI fragments I3 and K3 of the 'highly oncogenic' strain RB1B of MDV. The entire nucleotide sequence of the gene has been determined and its predicted amino acid sequence shown to share gross overall structural features with the gB genes of HSV, varicella-zoster virus (VZV) and other mammalian herpesviruses. In particular, all 10 cysteine residues were conserved in MDV gB and there was extensive homology throughout the gene with VZV, HSV and pseudorabies virus except for the N and C termini. The overall percentage amino acid identity between MDV gB and gB of the alphaherpesviruses had a mean of 50% which was almost twice that between cytomegalovirus and Epstein-Barr virus. Northern blot analysis showed that the main RNA transcribed from this gene is approx. 2.7 kb in size. Antibodies raised against synthetic peptides (residues 250 to 271 and 304 to 330) allowed the identification of a family of serologically related glycoproteins of Mr 110K, 64K and 48K in extracts of MDV-infected cells using immunoblots. Furthermore, the antisera were able to differentiate between the antigens of MDV and herpesvirus of turkeys in immunoblots. Immunofluorescence tests indicated that MDV gB is associated with granules in the cytoplasm and is present at the surface of MDV infected cells. PMID- 2544667 TI - Expression of glycoprotein D of herpes simplex virus type 1 in a recombinant baculovirus: protective responses and T cell recognition of the recombinant infected cell extracts. AB - Recombinant baculoviruses expressing glycoprotein D (gD) of herpes simplex virus type 1 (HSV-1) have been generated. The proteins expressed from the recombinants have been characterized using monoclonal antibodies on Western blots and by immunoprecipitation. Partially glycosylated 48K polypeptides have been identified as products of the gD gene. Polyclonal sera from H-2k mice infected with HSV-1 recognized the same polypeptides. Furthermore, draining lymph node cells from H 2k mice infected with HSV-1 proliferated in vitro in response to recombinant infected cell extracts. Immunization with such extracts generated high titre complement-dependent and -independent neutralizing antibody and the mice were protected against a challenge with HSV-1. PMID- 2544668 TI - Enhancement of virus transmission by tick salivary glands. AB - Previous studies have demonstrated that Thogoto (THO) virus is transmitted from infected to uninfected ticks cofeeding on an uninfected guinea-pig, although the guinea-pig does not develop a detectable viraemia. To investigate this mode of transmission, guinea-pigs were infected with uninfected Rhipicephalus appendiculatus nymphs prior to inoculation with either a mixture of THO virus and tick salivary gland extract, or with THO virus alone. The number of ticks that acquired the virus from feeding on animals inoculated with a mixture of virus and salivary gland extract was 10-fold greater than the number that became infected by feeding on animals inoculated with virus alone. The increase in the number of ticks that became infected was greatest when the salivary glands used in the inoculum were derived from uninfected ticks, which had partially fed for a period of 6 days. Viraemia was not detected in any of the guinea-pigs tested during the experiments. These results indicate that THO virus transmission is enhanced by factor(s) associated with the salivary glands of ticks, and that these factor(s) may facilitate 'non-viraemic' transmission between infected and uninfected ticks. PMID- 2544670 TI - Persistent replication of herpes simplex virus type 1 in JOK-1 cells. AB - Infection of the human B cell line JOK-1 with herpes simplex virus type 1 persisted over a period of more than 12 months (to date). Although limited cytopathic effects were seen, viral infection did not lead to extinction of the culture. Infectious centre assays, performed at various times after infection, revealed that only a small proportion of cells (1 to 10%) produced infectious virus particles. However, immunofluorescence studies showed that at any given time considerably more cells than calculated by infectious centre assays contained the immediate early viral protein ICP4 and expressed viral glycoproteins. These observations were confirmed by in situ hybridization analyses which revealed the presence of viral DNA even in cells not producing infectious particles. Since no evidence for the involvement of interferon could be found, some other so far unknown intrinsic property of the cells must be responsible for the restriction of virus replication and/or maturation. PMID- 2544669 TI - Studies on the mechanism of the interferon-mediated antiviral state to vesicular stomatitis virus in resting mouse peritoneal macrophages. AB - We have analysed the expression of vesicular stomatitis virus (VSV) proteins in virus-infected freshly explanted mouse peritoneal macrophages (resistant to virus replication), macrophages aged in vitro (permissive for virus replication) and freshly explanted macrophages from mice treated with antibody to interferon (IFN) alpha/beta (permissive for VSV replication). Our data showed that some VSV proteins (i.e. N/NS and G) were synthesized in virus-infected (1 p.f.u/cell) freshly harvested macrophages at early times after infection (3 to 6 h); the expression of such viral proteins was subsequently inhibited at 18 h post infection. In contrast, a progressive increase in the expression of VSV proteins was observed in the macrophages aged in vitro and infected with VSV at 1 p.f.u./cell. Infection with a higher m.o.i. (16 p.f.u./cell) resulted in similar viral protein electrophoresis patterns for both aged macrophages and freshly explanted macrophages. Even at low m.o.i. a marked and progressive expression of all VSV proteins was observed in freshly harvested macrophages from mice treated with antibody to mouse IFN-alpha/beta. Higher levels of oligo-2',5'-adenylate synthetase (2-5AS) were found in freshly harvested macrophages than in either aged macrophages or those from mice treated with antibody to IFN. No dsRNA dependent 67K protein kinase was detected in freshly harvested macrophages or peritoneal cells from untreated mice or mice treated with poly(rI).poly(rC) or Newcastle disease virus. The following conclusions can be drawn from these results. Low levels of spontaneous IFN-alpha/beta are responsible for the time dependent inhibition of VSV protein synthesis in virus-infected freshly harvested macrophages; high levels of 2-5AS (in the absence of detectable levels of 67K protein kinase) appear to correlate with the progressive inhibition of VSV proteins; this natural antiviral state is highly effective only at low m.o.i. PMID- 2544671 TI - Structure and expression of an integrated human papillomavirus type 16 genome amplified in a cervical carcinoma cell line. AB - A cellular sequence containing the integrated human papillomavirus type 16 genome in a cervical carcinoma cell line QG-U was cloned and analysed. The transcriptionally active viral genome disrupted at the E2 and L2 open reading frames was amplified with its flanking sequences. PMID- 2544672 TI - Retrovirus-induced cell fusion is enhanced by protease treatment. AB - The effect of proteases on retrovirus-induced cell fusion was studied. Both 'fusion from within' (fusion between infected and uninfected cells) and 'fusion from without' (fusion of uninfected cells by virus addition) was enhanced after treatment with proteases. An enhancement of up to 10-fold, giving 30% fusion of the cells, was observed. Protease treatment of infected cells or of virus particles enhanced fusion, indicating that cleavage of the virus surface proteins is important for cell fusion. Cell fusion is believed to reflect the virus-cell membrane fusion which occurs during the entry of enveloped viruses into cells. It is therefore possible that proteolytic cleavage of virus surface components during entry is involved in virus infection. PMID- 2544673 TI - Infection of rats with bovine leukaemia virus: establishment of a virus-producing rat cell line. AB - Adult rats were infected with bovine leukaemia virus (BLV). Inoculated rats persistently produced antibodies directed against viral structural proteins. No major pathogenesis in infected rats was found during 2 years of observation. It was possible to recover the virus from rat spleen several months after infection. A cell line, R(BLV), was established from rat spleen; this contained integrated BLV provirus. R(BLV) cells kept for over 80 passages in vitro produced viral particles with the properties of BLV. Provirus reintegration and/or amplification occurred in R(BLV) cells. The cell line was found to be tumorigenic in rats, and the virus produced was immunogenic. R(BLV) cells represent the first described BLV-producing rat cell line. Proven persistent infection with BLV indirectly suggests that rats can serve as a reservoir of BLV in nature. PMID- 2544674 TI - Antibody against Epstein-Barr virus DNA polymerase activity in sera of patients with nasopharyngeal carcinoma. AB - A salt-dependent DNA polymerase activity was demonstrated in the culture of an EBV-producing, lymphoblastoid cell line (NPC-204 cells) treated with 5-iodo-2' deoxyuridine (IUdR). There was a high frequency of levels of antibody to this enzyme in sera of patients with nasopharyngeal carcinoma (NPC). In contrast, sera from healthy subjects had little or no neutralizing activity. The high antibody level appeared as early as stage 1 of the disease in many NPC patients. The levels of the antibody increased with the progression of the disease and declined in treated patients. The results strongly suggest that tests measuring serum antibody against EBV DNA polymerase activity can be used for early diagnosis and prognosis of NPC. PMID- 2544675 TI - Brief report: killer cell defect and persistent immunological abnormalities in two patients with chronic active Epstein-Barr virus infection. AB - Two members of a family have manifested a syndrome of chronic active Epstein-Barr virus (EBV) infection. A father and his daughter suffered prolonged or recurrent mononucleosis, with splenomegaly, anemia, and intermittent fever; persistent immunological abnormalities included defective natural killer (NK) cytotoxicity, inverted CD4/CD8 ratios, hyper IgG1, high EBV viral capsid antigen (VCA) and early antigen (EA) antibodies, and low or undetectable EBV nuclear antigen (EBNA) antibody titers. The EBV seronegative member of the family was free of these abnormalities. However, NK activity in the seronegative individual was low-normal and its EBV-specific antibody-dependent K-cell cytotoxicity (EBV-ADCC) was abnormally low, suggesting that this K-NK cell defect may be primary. The father, who suffered from the syndrome for more than 15 years, lacked (or lost) antibodies to EBV-envelope and infected cell membranes, such as antibody dependent cellular cytotoxicity (ADCC), neutralizing (NT), and gp 350/220 antibodies. Slow improvement over a period of years was heralded by rising NK cytotoxicity. PMID- 2544676 TI - Serological typing scheme for BK-like isolates of human polyomavirus. AB - Human polyomavirus BK-like isolates were subjected to restriction endonuclease analysis with the enzymes EcoRI and Hind III. End-point dilution was used to obtain homogeneous virus pools for DNA analysis and to remove JC virus from a mixed stock. The results of Hind III digestion suggested that two subgroups could be distinguished. Several BK-like isolates were purified and rabbit antisera raised. The isolates were compared with each other and with BK and JC viruses by haemagglutination-inhibition (HI) and by neutralisation. JC virus was serologically distinct, but all the other isolates showed some cross-reactivity. Two subgroups were again evident: GS and PG were with prototype BK in subgroup 1, and MG and IV were in subgroup 2. Two isolates, AS and SB, reacted with isolates of both subgroups 1 and 2 but were distinct from one another: their genome was similar to subgroup 1 isolates. Typing by HI or by neutralisation may form a basis for grouping BK-like polyomavirus isolates. PMID- 2544677 TI - Characterization of two genome types of adenovirus type 31 isolated in Stockholm during 1987. AB - The authors isolated during 1987 seven adenovirus type 31 (Ad31) within a 9-month period. The isolates were obtained from urine, throat, and feces, implying a systemic spread of the infection. Most patients displayed gastrointestinal symptoms, but some had respiratory symptoms and fever. All of the strains differed by restriction endonuclease analysis from the prototype strain (1315) by an additional Bgl II restriction site. Ad31 isolates 1-6 could be divided into two groups by the enzymes Bam HI, Msp I, and Xho I. Each enzyme gave rise to the same group distribution: isolates 1-3 and 4-6, respectively. Digestion with Bst EII, Hind III, Kpn I, and Sma I resulted in identical patterns for isolates 1-6. Isolate 7, however, demonstrated a DNA deletion of approximately 0.8 kbp, but it was otherwise identical to isolates 4-6. In conclusion, two separate genome types of Ad31 were isolated, one of which included a DNA deletion mutant. The increased isolation rate may reflect an epidemiological situation, as the same isolation procedure had been used both before and after this period. PMID- 2544678 TI - Brief report: primary human herpesvirus 6 infection in a patient following liver transplantation from a seropositive donor. AB - A virus, isolated from peripheral blood lymphocytes of a patient 23 days after liver transplantation, was identified by negative contrast and thin-section electron microscopy, DNA hybridisation, and immunofluorescence as human herpesvirus 6 (HHV-6). The patient was seronegative for antibodies to HHV-6 before receiving the liver from a seropositive donor. Seroconversion for IgM and IgG antibodies occurred at the same time as the viraemia, indicating a primary response to HHV-6 infection that was probably acquired from the donor liver. The relationship of the infection to the patient's subsequent disease episodes is discussed. PMID- 2544679 TI - Amplification of rhinovirus specific nucleic acids from clinical samples using the polymerase chain reaction. AB - We describe a novel method for the detection of human rhinoviruses in clinical samples, using the polymerase chain reaction. Two synthetic oligonucleotide primers were produced that bind in the 5' noncoding region of all rhinovirus serotypes tested, about 350 nucleotides apart, and were used to prime polymerase chain reaction amplification of the intervening stretch of DNA. The product of this reaction, which can be clearly visualized by gel electrophoresis, is a discrete 380 bp band, the occurrence of which is diagnostic of the presence of a rhinovirus in the clinical sample analysed. The technique, which is rapid, sensitive, and reliable, has been used successfully for all the different rhinovirus serotypes tested to date in our laboratory. However, the sensitivity of detection is greatly dependent on the inclusion of both tRNA and vanadyl complexes during the viral RNA extraction process. Using this technique, under optimal conditions, we were able to detect virus in clinical samples with titres as low as TCID50 10(2.5). PMID- 2544681 TI - Induction and persistence of local rotavirus antibodies in relation to serum antibodies. AB - The induction and persistence of local rotavirus antibodies, including stool IgA, jejunal IgA, and jejunal neutralizing antibody, were evaluated in 14 adult volunteers infected with the CJN strain of human rotavirus. In addition, the relationships between local rotavirus IgA and serum rotavirus IgA, IgG, and neutralizing antibody were determined. Both stool and serum rotavirus IgA appeared to have similar kinetics. Both antibodies peaked by days 14-17 after inoculation in all subjects, then decreased rapidly. By days 26-28, titers had fallen to 13% and 30% of their respective peaks. Serum rotavirus IgG peaked somewhat later, occurring in five subjects on days 26-28. Serum neutralizing antibody peaked on days 26-28 in all but three subjects. Both serum IgG and neutralizing antibodies also declined more slowly than rotavirus IgA. Although all antibody concentrations had decreased to only a fraction of their peak responses by days 270-365 after rotavirus inoculation they remained higher than baseline levels. For example, stool rotavirus IgA concentrations were 13.5-fold higher than baseline, while jejunal rotavirus IgA and neutralizing antibody were 8.9- and 4.3-fold above baseline, respectively. Similarly, serum antibodies remained 3.7- to 11.2-fold higher than baseline at 270-365 days after rotavirus inoculation. These studies imply that serum rotavirus IgA is a good indicator of local antibody responses. Furthermore, although both serum and local antibody titers peaked within 2-4 weeks after infection, these antibodies persisted at above baseline concentrations for at least 9-12 months after infection. PMID- 2544680 TI - Shedding of herpes simplex virus type 1 into saliva in patients with orofacial fracture. AB - Shedding of herpes simplex virus type 1 (HSV-1) into saliva was studied in 83 patients with orofacial fractures. Infectious virus was isolated from 14 of 83 patients (16.8%) with no detectable herpetic lesion during hospitalization. Of the 83 patients, 44 (53%) had HSV-1 specific antibody. Virus shedding into saliva was observed only in the patients with antibody to HSV-1; thus, the frequency of virus shedding patients among those with antibody was 31% (14 of 44 patients). The frequency was obviously higher than that of a healthy population. The period of HSV-1 shedding had a mean of 3.7 days with range of 1 to 8 days, which is also significantly longer than that of a healthy population. These results strongly suggest that both treatment and operation lead effectively to reactivation of latently infected HSV-1. PMID- 2544682 TI - Abnormalities of sympathetic vasomotor tone in distal axonal neuropathy. AB - Microneurographic studies have shown that sympathetic vasomotor nerve activity is observed less frequently in patients with peripheral neuropathy than in controls, but when detected its pattern and the baroreflex latencies appear to be normal. Vasomotor nerve function was examined in chloralose-urethane anaesthetised dogs by comparing the discharge of renal sympathetic nerves in control animals and animals with acrylamide neuropathy. There was a normal pattern of pulse related inhibition of sympathetic nerve activity as well as normal amplitude spontaneous compound nerve action potentials in the animals with neuropathy. When vasomotor tone was altered abruptly by raising pressure in a bilateral isolated carotid sinus preparation, the baroreflex latencies were normal in the affected animals. However, when carotid sinus pressure was kept constant, pulse-related sympathetic inhibition, normally mediated by vagal cardiopulmonary baroreceptors, was absent in animals with neuropathy. It is likely that the vagal nerve fibres to cardiopulmonary baroreceptors as well as the receptors themselves are damaged while shorter carotid sinus nerve fibres are relatively spared in axonal neuropathies. As long as these shorter nerves are intact patients with axonal neuropathy should have relatively normal baroreflexes and normal sympathetic vasomotor tone when measured microneurographically. PMID- 2544683 TI - Electron spin resonance studies of erythrocyte membrane in spinocerebellar degeneration. AB - Erythrocyte membrane fluidity was examined by electron spin resonance spectra using nitroxide fatty acid spin labels in spinocerebellar degeneration (SCD). Subjects with SCD, motor neuron disease (MND) and controls did not differ in fluidity of the deep site (hydrophobic region) of the erythrocyte membrane. However, the fluidity of the shallow site (hydrophilic region) in the erythrocyte membrane was significantly less fluid in SCD than in controls and MND (outer hyperfine splitting of 5-nitroxide stearic acid: SCD 54.70 +/- 0.43 G, controls 53.57 +/- 0.41 G, MND 53.54 +/- 0.35 G, P less than 0.001). Serum HDL-cholesterol and membrane fluidity correlated significantly in controls, but not in SCD. A significant negative correlation between age and membrane fluidity was found in SCD, but not in controls. These data suggest that membrane abnormality exists in SCD and may be concerned with aging. PMID- 2544684 TI - Hypertension in long-term survivors of childhood renal cancers. AB - The prevalence of hypertension was investigated in 119 adults who have survived for up to 53 years following the diagnosis of renal cancer in childhood (Wilms' tumor, 116 patients; renal carcinoma, three patients). Twenty-four (20%) have developed definite or borderline hypertension, as compared with 18.1 cases expected based on US population rates (relative risk [RR], 1.3; 95% confidence interval [CI], 0.9 to 2.0; P = .20). This nonsignificant excess is due to the heightened prevalence of definite hypertension among one subgroup of male patients. The findings are not explained by cigarette smoking, obesity, age, and stage at diagnosis of Wilms' tumor, or family history of hypertension. A case comparison analysis within the cohort showed no consistent hypertensive effect associated with radiation therapy dose, radiotherapy concurrent with dactinomycin chemotherapy, or extent of renal surgery. Hypertension is not a common late complication of Wilms' tumor in our patients. PMID- 2544685 TI - Primary chemotherapy of brain metastasis in small-cell lung cancer. AB - Fourteen patients with brain metastases from previously untreated small-cell lung cancer (SCLC) were treated with three courses of systemic chemotherapy as an initial mode of treatment. Whole brain irradiation was given concurrently with the fourth course of chemotherapy. The chemotherapy consisted of cyclophosphamide, 600 mg/m2 intravenously (IV) on day 1; doxorubicin, 50 mg/m2 IV on day 1; vincristine, 1.5 mg IV days 1 and 5; and etoposide, 60 mg/m2 IV days 3 through 5; all repeated every 3 weeks with dosage adjustments. There were ten men and four women, with a median age of 59 years (range, 47 to 75). Six patients had multiple brain lesions, and the brain was the sole site of distant metastasis in four patients. Three patients were inevaluable for response in the brain, as two died early and the third dropped out of the trial too soon. Brain lesions responded to chemotherapy in nine (one complete remission [CR], eight partial remissions [PR]) of 11 (82%) evaluable patients, and objective responses in the extracranial lesions were documented in nine (one CR, eight PR) of 12 (75%) evaluable patients. Median survival was 34 weeks (range, 1 to 93), and two patients are still alive. Toxicity was significant, with severe granulocytopenia (less than 500/microL) and thrombocytopenia (less than 50,000/microL) observed in 85% and 15% of patients, respectively. Six patients had major infectious complications, which resulted in septic deaths in two. However, there was no deterioration of neurologic status during the initial phase of treatment with chemotherapy. We conclude that systemic chemotherapy alone can induce objective regression of metastatic brain lesions in patients with previously untreated SCLC. PMID- 2544686 TI - Lack of relationship between in vitro tumor cell growth and prognosis in extensive-stage small-cell lung cancer. AB - The ability to establish a continuously growing tumor cell line from fresh tumor specimens has been associated with shortened survival in some human malignancies. Therefore, we assessed the relationship between survival and in vitro tumor cell growth from specimens obtained during routine staging procedures in 68 consecutive patients with untreated, extensive-stage small-cell lung cancer (SCLC) who received etoposide/cisplatin chemotherapy. Three groups of SCLC patients could be distinguished: (1) 23 patients in whom a tumor cell line was established in vitro; (2) 28 patients in whom tumor-containing specimens were cultured but in vitro growth did not occur; and (3) 17 patients in whom no tumor containing specimen could be procured. No significant difference in response rates to chemotherapy of the three groups was noted. Poor performance status (P2 = .001), male gender (P2 = .0008), liver metastases (P2 = .0033), brain metastases (P2 = .0152), and the ability to obtain a tumor-containing specimen from the patient for laboratory culture (P2 = .0005) were all significant independent predictors of decreased survival in this patient population. While the ability to obtain a tumor cell specimen for cell culture using routine staging and diagnostic procedures identified patients with shortened survival, we found no significant survival differences between patients whose tumor cell specimens grew in cell culture v those that did not (median survival of 7 months v 11 months, P2 = .72). Our study indicates that the clinical outcome of extensive-stage SCLC patients from whom tumor cell lines can be established is not significantly different than in those cases from whom tumor-containing specimens could not be grown in vitro. PMID- 2544687 TI - Dose-intensive chemotherapy in refractory germ cell cancer--a phase I/II trial of high-dose carboplatin and etoposide with autologous bone marrow transplantation. AB - Between September 1986 and March 1988, 33 patients with refractory germ cell cancer were entered on a phase I/II trial of two courses of high-dose carboplatin plus etoposide with autologous bone marrow support. All patients had extensive prior treatment and had either cisplatin-refractory disease (67%) defined as progression within 4 weeks of the last cisplatin dose or failed at least two cisplatin-based regimens (35%) including a cisplatin-ifosfamide salvage regimen. Patients received a fixed total dose of etoposide of 1,200 mg/m2 with each cycle. The carboplatin dose ranged from 900 mg/m2 to 2,000 mg/m2. Twenty of the 33 patients received the second cycle of therapy. Despite extensive prior therapy with cisplatin, neurotoxicity, nephrotoxicity, or hearing impairment with high dose carboplatin and etoposide was unusual. The most common nonhematologic toxicity was moderate enterocolitis. The hematologic toxicity of this regimen was substantial at each dose level. All 53 courses were accompanied by granulocytopenic fevers. Seven of the 33 patients (21%) died from treatment. All of these deaths occurred during the granulocyte nadir, and five were related to documented sepsis. Overall, 14 of 32 patients (44%) evaluable for response obtained an objective response, including eight complete remissions. Four patients remain in complete remission, with three patients being continuously free of disease in excess of 1 year. Eight responders (including four complete remissions) had progressed while receiving cisplatin. We conclude that carboplatin and etoposide can be administered in combination at high dosages and this regimen may have curative potential for patients with germ cell tumors resistant to conventional-dose cisplatin-based therapies. PMID- 2544688 TI - Comparison of intermittent versus continuous infusion metoclopramide in control of acute nausea induced by cisplatin chemotherapy. AB - Sixty previously untreated patients with newly diagnosed advanced-stage lung cancer (21 small-cell, 39 non-small-cell) received chemotherapy with cisplatin and etoposide. Bleomycin was also used in the patients with non-small-cell lung cancer. During the first cycle of chemotherapy, 30 patients received antiemetic therapy with intermittent metoclopramide (regimen A), and the other 30 patients received continuous infusion metoclopramide (regimen B). During the second course of chemotherapy, patients were switched to the alternate regimen. Regimen A consisted of lorazepam, 1 mg, orally; dexamethasone, 10 mg, intravenously (IV) every four hours for three doses; diphenhydramine, 0.5 mg/kg, IV every four hours for three doses; metoclopramide, 1 mg/kg, IV bolus every two hours for six doses. Regimen B was identical to A except metoclopramide was administered as 1 mg/kg, IV bolus followed by 0.5 mg/kg/h for ten hours. Fifty-eight patients completed both antiemetic regimens. Thirty-nine of the 58 patients had total control of acute nausea and vomiting (0-1 episodes) with regimen A or B. Fourteen patients had poor control of acute nausea and vomiting (more than one episode) with regimen A but total control with regimen B. Five patients had poor control with either regimen. Dystonic reactions, akathisia, or diarrhea occurred in 20 of the 58 patients on regimen A, but in only eight of the 58 patients on regimen B. Compared with intermittent bolus, continuous infusion metoclopramide is more effective in total control of acute nausea and vomiting and has less toxicity. PMID- 2544689 TI - Imaging of brain tumors using peripheral benzodiazepine receptor ligands. AB - Peripheral benzodiazepine receptor ligands were utilized to selectively image intracerebrally implanted C6 gliomas, RG-2 gliomas, and Walker 256 metastatic tumors by means of quantitative autoradiography. Intravenous injections of 3H PK11195 (1-(2-chlorophenyl)-N-methyl-N-(1-methylpropyl)-3-isoquinoline carboxamide) or 3H-flunitrazepam in combination with clonazepam revealed high densities of peripheral benzodiazepine binding in glial tumors, with less binding in metastatic tumors. Peripheral binding was displaced by preadministration of excess PK11195. Topographical correlation was excellent between areas of histologically verified tumor and high densities of peripheral benzodiazepine binding. The choroid plexus, ependyma, and pineal gland also showed a moderate level of binding, but there was little binding in other normal brain structures or necrotic tissue. Binding densities were three- to fivefold higher in C6 glial tumors compared to normal cortex. Injection of 3H-flunitrazepam alone, which binds to both central and peripheral receptors, had the advantage of showing normal anatomic structures in addition to a clear definition of tumor topography. The potential value of peripheral benzodiazepine ligands in selectively imaging brain tumors in man with positron emission tomography is discussed. PMID- 2544690 TI - Technetium-99m (v) dimercaptosuccinic acid uptake in patients with head and neck squamous carcinoma: experience in imaging. AB - A recently developed imaging agent, technetium-99m (v) dimercaptosuccinic acid (99mTc (v) DMSA), has been used to assess head and neck squamous carcinoma (SCC). We have prospectively studied 62 patients of whom 53 had a histologically proven head and neck SCC. The remaining nine had benign lesions. The results of planar imaging in patients with primary disease yielded an 85% sensitivity and 78% specificity. Planar imaging in patients with cervical lymphadenopathy revealed a 59% sensitivity. Nineteen patients also had single photon emission computed tomography imaging which improved the image quality, spatial resolution and sensitivity of the investigation. Twenty-seven patients were scanned before and after radiotherapy and, of these, 96% showed positive uptake in the salivary glands with no evidence of tumor recurrence. This study has shown 99mTc (v) DMSA imaging provides a cheap and rapid method of investigating head and neck SCC and further studies are necessary to evaluate its role in the management of patients with this disease. PMID- 2544691 TI - Scintigraphic and ultrasound features of giant hemangiomas of the liver. AB - Giant hemangiomas of the liver are clinically distinct from smaller and more innocent hemangiomas as they are more prone to complications. On imaging with ultrasound, they can also be readily confused with hepatoma, metastatic disease, or focal nodular hyperplasia. Nine giant hemangiomas (maximal diameter greater than 8 cm) were studied by scintigraphy and ultrasound. In all instances, the fill-in of the lesion on [99mTc]RBC scintigraphy indicated the diagnosis of hemangioma, adding specificity to the screening sonographic study. The pattern of fill-in on scintigraphy also appeared to be size dependent with lesions less than 11 cm in diameter equilibrating uniformly, while larger abnormalities intensified in centripetal fashion. PMID- 2544692 TI - Quantitation of renal uptake of technetium-99m DMSA using SPECT. AB - Quantitative single photon emission computed tomography (SPECT) methodology based on calibration with kidney phantoms has been applied for the assessment of renal uptake of [99mTc]DMSA in 25 normals; 16 patients with a single normal kidney; 30 patients with unilateral nephropathy; and 17 patients with bilateral nephropathy. An excellent correlation (r = 0.99, s.e.e. = 152) was found between SPECT measured concentration and actual concentration in kidney phantoms. Kidney uptake at 6 hr after injection in normals was 20.0% +/- 4.6% for the left and 20.8% +/- 4.4% for the right. Patients with unilateral nephropathy had a statistically significant (p less than 0.001) low uptake in the diseased kidney (7.0% +/- 4.7%), but the contralateral kidney uptake did not differ from the normal group (20.0% +/- 7.0%). The method was especially useful in patients with bilateral nephropathy. Significantly (p less than 0.001) decreased uptake was found in both kidneys (5.1% +/- 3.4% for the left and 6.7% +/- 4.2% for the right). The total kidney uptake (right and left) in this group showed to be inversely correlated (r = 0.83) with serum creatinine. The uptake of [99mTc]DMSA in single normal kidney was higher (p less than 0.001) than in a normal kidney (34.7% +/- 11.9%), however, it was lower than the total absolute uptake (RT + LT = 41.5% +/- 8.8%) in the normal group. The results indicate that SPECT is a reliable and reproducible technique to quantitate absolute kidney uptake of [99mTc]DMSA. PMID- 2544693 TI - Chemical aspects of labeling sucralfate with 99mTcO4. AB - Two formulations of [99mTc]sucralfate have been used to image gastric and duodenal ulcers and inflammatory bowel disease. One formulation is a complexation of [99mTc]HSA with sucralfate. The second is prepared by directly labeling sucralfate with [99mTc]pertechnetate in the presence of stannous ion. An in vitro study of the factors affecting the production and stability of these labeled sucralfate preparations was conducted. Both formulations were stable at the acidic pH likely encountered in the stomach. However, at pH greater than 6 the albumin-sucralfate complex began to dissociate while directly labeled sucralfate was stable to a pH of 9. Conversely it was shown that directly labeled sucralfate was more susceptible to loss of 99mTc to other chelating species. Sucralfate complexed with [99mTc]HSA was radiochemically stable up to a specific activity of 26 GBq (700 mCi) per gram while directly labeled sucralfate showed decreased 24 hr stability at specific activities greater than 837 mCi (31 GBq) per gram. PMID- 2544694 TI - Curative radioimmunotherapy of human mammary carcinoma xenografts with iodine-131 labeled monoclonal antibodies. AB - The radioiodinated monoclonal antibody BW 495/36 showed an exceptionally high uptake and long residence time in human ductal mammary carcinoma xenografts in nude mice. There was a mean tumor uptake of 82%/g 24 hr p.i., decreasing with a biologic half-life of approximately 6 days, to 15%/g by Day 16. The tumor-to blood ratio increased from 2.8 to 21.4 and the percentage of the whole-body retention recovered in the tumor from 47% to 80% during the same time interval. The therapeutic efficiency of two injections of 7.4 MBq 131I-BW 495/36 was evaluated by comparing the tumor size with that in mice injected with either the same amount of the unlabeled MoAb, the same radioactivity of an 131I-labeled nonspecific MoAb, or with saline only. The high tumor accumulation of 131I-BW 495/36 led to a total tumor dose of 77 Gy resulting in a mean reduction in tumor diameter of 50%, corresponding to a reduction in tumor volume of 88% within 42 days p.i. Unlabeled MoAb had no effect on tumor growth compared with controls, whereas 131I nonspecific antibody caused a slight inhibition of tumor growth. Histologic tumor sections showed large areas of necrosis and a pronounced vacuolation of the tumor cell cytoplasm between Days 7 and 30 p.i. By Day 42 all remaining tissue in the tumor was identified as mouse connective tissue. PMID- 2544695 TI - Osmium-191/iridium-191m generator based on silica gel impregnated with tridodecylmethylammonium chloride. AB - A new separation system for an 191Os/191mIr generator is described. The system is composed of two columns in a series: a main column, packed with silica gel impregnated with tridodecylmethylammonium chloride (loaded with high activity 191Os as an osmyl chloride); and a scavenger column, packed with activated charcoal. Iridium-191m is eluted from the generator by pH 1 saline. For clinical use the eluate is buffered by succinate solution before injection. This new system is characterized by high performance (approximately 25% 191mIr recovery with 5 X 10(-4)% 191Os breakthrough) and long shelf-life (3 wk). The buffered eluate is sterile, pyrogen-free, and nontoxic, and contains no 192Ir. It is suitable for first-pass radionuclide angiocardiography with a very low radiation dose to the patient. PMID- 2544696 TI - A realistic dynamic cardiac phantom for evaluating radionuclide ventriculography: description and initial studies with the left ventricular chamber. AB - A phantom was devised to validate scintigraphically determined left ventricular ejection fractions (LVEFs) and cardiac chamber volumes in the following simulated cardiac situations: normal contraction, moderately impaired left ventricular contraction, severely impaired left ventricular contraction, mitral regurgitation, and cardiomyopathy. The phantom, assembled from anatomically realistic cardiac chambers, simulated contraction and expansion using individual chamber pumps coordinated by a microcomputer. Scintigraphic studies were performed by sequential imaging of [99mTc]pertechnetate introduced into each chamber. The images were analyzed like conventional clinical studies, using both automatic and manual techniques. Scintigraphic techniques correlated with chamber volumes that were determined by weight to yield the following regression formulae: LVEF (by automatic method 1) = 1.08 x LVEF (by weight) -5.11; LVEF (by automatic method 2) = 1.00 x LVEF (by weight) -3.15; and LVEF (by manual method) = 1.04 x LVEF (by weight) -5.08 ml (Correlation coefficients greater than 0.98). The absolute left ventricular volumes (LVVs), determined by scintigraphy, correlated well with LVVs determined by weight. These correlations were performed with separations between the center of the left ventricle and the collimator varying from 5 cm to 9 cm. The regression formulae for 5, 7, and 9 cm distances were: LVV (by counts) = 0.99 x LVV (by weight) + 0.13, LVV (by counts) = 1.04 x LVV (by weight) + 9.08, LVV (by counts) = 0.88 x LVV (by weight) + 15.25, respectively. At 9 cm, slight volumetric underestimation occurred, as predicted from the work of Fearnow et al., possibly because of oversubtraction of background. Thus, this phantom provides a useful tool for validating scintigraphic cardiac blood-pool studies simulating a wide range of clinically relevant situations. PMID- 2544697 TI - Radiation absorbed dose from technetium-99m-labeled bone imaging agents. Task Group of the Medical Internal Radiation Dose Committee, The Society of Nuclear Medicine. PMID- 2544698 TI - Thallium-201 single photon emission computed tomography in the evaluation of suspected lung cancer. AB - Thallium-201 SPECT was performed in 30 patients with suspected lung cancer. Both early and delayed scans demonstrated abnormal accumulation in all of 23 malignant pulmonary lesions including 21 lung cancer and in two of seven benign conditions. There were significant differences in delayed ratio (uptake ratio of the lesion to the normal lung on delayed scan) and retention index (degree of retention in the lesion) between lung cancer and benign conditions, respectively (p less than 0.01, p less than 0.05). The delayed ratio and retention index revealed that adenocarcinoma showed higher 201Tl accumulation than squamous cell carcinoma and small cell carcinoma (p less than 0.05) and 201Tl clearance in squamous cell carcinoma was faster than in the other two (p less than 0.05). Mediastinal involvement was detected in five of seven patients on delayed scans. The smallest lesion depicted was 1.5 cm in diameter. Two false negatives had small metastases less than 1.0 cm in diameter. This method seems to be useful to detect lung cancer, to differentiate malignant from benign lesions, and to evaluate mediastinal involvement from lung cancer. PMID- 2544699 TI - Renal handling of technetium-99m DMSA: evidence for glomerular filtration and peritubular uptake. AB - The finding of an enhanced excretion of [99mTc]dimercaptosuccinic acid (DMSA) in patients with tubular reabsorption disorders prompted us to investigate the role of filtration in the renal handling of [99mTc]DMSA. Our studies in human serum indicated that binding to serum proteins was approximately 90%. Chromatography of human urine and studies in rats showed that the complex was excreted unaltered into the urine. Renal extraction of [99mTc]DMSA in a human volunteer was 5.8%. Continuous infusion of [99mTc]DMSA in 13 individuals with normal renal function gave the following results (mean +/- s.d.): plasma clearance of [99mTc]DMSA 34 +/ 4 ml/min, urinary clearance of [99mTc]DMSA 12 +/- 3 ml/min. The calculated filtered load of [99mTc]DMSA closely resembled the urinary clearance, whereas the plasma clearance was about three times faster. This indicates that peritubular uptake accounts for approximately 65% and filtration for approximately 35% of the renal handling of [99mTc]DMSA. PMID- 2544701 TI - Individualizing the nursing curriculum. PMID- 2544702 TI - The urgency of substance abuse education in schools of nursing. AB - The abuse of alcohol and drugs has become a major health and social problem in the United States. Nurses comprise the largest segment of health-care professionals, yet report their educational experiences offer little to prepare them to develop substance abuse prevention and intervention programs. This article identifies factors that contribute to this educational gap, discusses challenges in planning and implementing substance abuse curricula, and offers specific guidelines for improving educational experiences. PMID- 2544700 TI - Technetium-99m DISIDA hepatobiliary agent in diagnosis of hepatocellular carcinoma, adenoma, and focal nodular hyperplasia. PMID- 2544703 TI - Nursing education component in master's programs. AB - The purposes of this study were to: a) describe the outcomes, content, and structure of master's programs in nursing in the functional area of teaching; b) describe related learning experiences of students; and c) identify trends in graduate nursing education in preparing students for the role of teacher. A self administered questionnaire developed by the investigators was mailed to all NLN accredited master's programs (N = 139). The instrument collected data on the nursing education component of the master's program. Ninety-two questionnaires were completed and returned, resulting in a response rate of 66.2%. Only 10 (10.9%) master's programs offered a major in nursing education. More common were minors, elective courses, and tracks in nursing education, as well as other similar curriculum patterns (N = 66). Fifteen (16.3%) master's programs offered no courses in nursing education. There was a statistically significant difference between the mean number of credits of nursing education courses required in programs offering a major (M = 11.4) compared with a minor (M = 8.8). More than half of the master's programs required nursing education courses in learning theory, curriculum development, instructional design, teaching methods, clinical teaching, various aspects of evaluation, and grading. Learning experiences in which students participated to prepare themselves for teaching roles included in the majority of programs: classroom teaching, clinical teaching (generally involving a practicum), clinical evaluation of students, development of a course/curriculum, and test construction. The research documents the continued emphasis on advanced nursing practice as the primary area of graduate study with preparation for teaching occurring through minors in the program.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544704 TI - Academic middle managers for baccalaureate nursing: work motivation and satisfaction. AB - Seventy-nine percent of academic middle managers for baccalaureate nursing reported that they did not plan to continue in their current management positions, or advance in academic leadership positions (George, 1981). This study examined the relationships between the job characteristics, a mediating variable "growth need strength", and the general job satisfaction and work motivation of academic middle managers for baccalaureate nursing. The sample was drawn from the population of academic middle managers for baccalaureate nursing in 126 colleges and universities across the United States that offer both baccalaureate and higher nursing degree programs. Sixty seven percent returned the Job Diagnostic Survey Short Form (JDSSF). The findings were interpreted through an examination of three analytical techniques: zero-order correlations, multiple regression, and multiple regression with interaction. Findings indicate that the dependent variable, "internal work motivation," can be measured in the population, but it cannot be predicted using the seven job characteristics measured by the JDSSF. The job characteristic autonomy was significant in predicting "general job satisfaction." General job satisfaction also proved to be some combination of security satisfaction and growth need satisfaction. both independent variables "autonomy" and "feedback from the job itself" were significant in predicting growth need satisfaction. As a mediating variable, "growth need strength" was not significant in predicting general job satisfaction in a linear model, nor was it significant when entered as an interactive term. However, the multiplicative model did increase, by four percent above the linear model, the amount of variance predictable in general job satisfaction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544705 TI - Aptitude, previous achievement, and cognitive style: relation to academic achievement in nursing courses of differing content. AB - The questions studied were whether a) cognitive style might predict differential success in nursing courses having a different specialty content focus and b) whether after the inclusion of aptitude measures and previous achievement measures FDI cognitive style would significantly increase the prediction of academic achievement in nursing courses of differing specialty content. Senior baccalaureate nursing students completed the Group Embedded Figures Test as a measure of field-dependence-independence (FDI) cognitive style. Aptitude and achievement scores were obtained from students' files and instructors' grade sheets. The findings were that FDI cognitive style is not a predictor of differential academic success. The junior nursing grade point average was the best predictor of achievement in senior level nursing courses. The prediction of academic achievement of nursing students has received researchers' attention over the years in an attempt to identify data that might aid in the establishment of selection criteria, in program and curriculum planning, and in the establishment of remediation or supportive programs for enrolled students who have been identified as high risk. In most studies, affective factors alone, such as personality characteristics and vocational interests, have not been found to be predictors of academic achievement. Most researchers have found that previous achievement or aptitude measures are the best predictors of nursing course achievement (Schwirian, 1976; Grant, 1986). While previous achievement and aptitude measures may be used in the selection process, they are not helpful in giving direction for planning teaching-learning experiences. Cognitive style might serve this purpose if it was found to be predictive of academic achievement.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544707 TI - The clinical teaching associate model: advantages and disadvantages in practice. AB - Two major benefits of this model are the increased availability of the CTA to the student and the LT's ability to observe the student's overall progress through clinical and classroom areas. In addition, the CTA Model supports the recent trend toward collaboration between schools of nursing and hospitals. Implementation of the CTA Model provides a unique opportunity for research into the effectiveness of such a model. The following are some questions that need to be addressed in evaluating the CTA Model: Is there greater patient, student, and/or instructor satisfaction with this model? Does the model allow faculty more time to devote to research and publication? Do students learn more under this model? The answers to these questions could provide valuable insights into the educational merit of the CTA Model. PMID- 2544706 TI - Wrestling with the larger picture: placing ethical behavior in clinical situations in context. AB - This article provides an account of the use of a model-building process as an educational strategy for the teaching of ethics. Designed to integrate students' growing knowledge and skill in nursing with their intellectual and professional development, this model-building process has its theoretical foundations in cognitive moral development theory, and in an integrative approach to nursing education called Multi-Course Sequential Learning (MCSL). PMID- 2544708 TI - Succinate dehydrogenase and synthetic pathways of glucose 6-phosphate are also the markers of the toxicity of orally administered secondary autoxidation products of linoleic acid in rat liver. AB - In order to find the markers of the toxicity of the autoxidized lipids in the liver, rats were given a lethal amount of secondary autoxidation products of linoleic acid (400 mg/rat/day for 3 days) and then changes in the hepatic metabolic functions were analyzed. A decrease in acetyl-CoA level to half caused by the depletion of CoASH was reported in an associated paper (J. Nutr. Sci. Vitaminol., 35, 11-23, 1989). Citrate, isocitrate, and 2-oxoglutarate also decreased to half the level of those of the control group. Reduction in isocitrate dehydrogenase activity was only 25%, while NADH2 and ATP levels remained unchanged. Thus, the reduction in the citrate cycle activity was due to the decrease in acetyl-CoA. The activity of mitochondrial succinate dehydrogenase was decreased to 1/5. Other appreciable changes were depletion of glucose 6 phosphate and fructose 6-phosphate, accumulation of glucose 1-phosphate, reductions in hexokinase, phosphofructokinase, glucose-6-phosphatase, phosphoglucomutase, and phosphogluconate dehydrogenase activities, and decrease in the NADPH2 level. It was considered that these changes were caused by the depletion of glucose 6-phosphate whose synthetic pathways were abnormal. Therefore, the markers of the hepatotoxicity of secondary products were the changes in the CoASH level and the activities of succinate dehydrogenase and synthetic pathways for glucose 6-phosphate. PMID- 2544709 TI - Injury of the anterior cruciate ligament: the role of collagenase in ligament degeneration. AB - Rapid degeneration of the anterior cruciate ligament (ACL) has been observed following acute ACL rupture. An understanding of this process might explain some of the poor clinical results of primary ACL repair. We created a surgical rabbit model of acute ACL injury and developed an in vitro assay for collagenase activity in the ACL and menisci. Microscopic evaluation revealed a rapidly degenerative process in injured ACLs, with loss of cellularity and matrix organization. This was associated with a significant increase in collagenase activity and a decrease in total collagen of the injured ACLs as compared with sham-operated controls. These findings confirm the observation that cut ACL ligament ends rapidly degenerate. This degenerative process might be partly due to a response of cells intrinsic to the ACL to injury. Left unchecked, this process may be detrimental to surgical attempts for primary ACL repair. PMID- 2544710 TI - Different effects of mechanical vibration on bone ingrowth into porous hydroxyapatite and fracture healing in a rabbit model. AB - The effects of mechanical vibration on bone ingrowth into porous hydroxyapatite implants and fracture healing were examined radiographically, histomorphometrically, and biomechanically in a rabbit model. Fifty-three female NZW rabbits were used in this study. These rabbits were divided into four separate studies to assess the effects of 20 and 60 min of vibration/day in both implant and osteotomy models as compared with the respective non-vibrated controls. For the implant model, coral hydroxyapatite goniopora rods were implanted bilaterally into tibiae and for the osteotomy model, bilateral fibulae were osteotomized. A resonant frequency of 25 Hz mechanical vibration was used. After periods of 2, 3, 4, and 6 weeks of vibration, the rabbits were killed and examined. For the implant model, there was no significant difference between control, 20, or 60 min of vibration/day with respect to the rate or amount of new bone ingrowth. For the fracture model, 60 min of vibration/day produced a significantly larger callus as compared with the non-vibrated controls (p less than 0.05), whereas 20 min of vibration/day did not. Although biomechanical testing demonstrated a general trend for increased strength in the vibrated animals, it failed to reach significance. These results suggest that the mechanical vibration used in the present study had a beneficial effect on callus volume, possibly due to the stimulation of secondary bone healing processes, but does not appear to promote bone ingrowth into a porous hydroxyapatite implant. PMID- 2544711 TI - Heterotopic osteogenesis in porous ceramics induced by marrow cells. AB - When untreated porous calcium phosphate ceramics were transplanted into subcutaneous (s.c.) or intramuscular (i.m.) sites, fibrovascular tissue grew in the pore region without evidence of bone formation. However, when these same ceramics were combined with syngeneic marrow cells, osteogenesis was observed inside the pore region of the implanted ceramic. The osteogenesis began on the surface of the pore region at approximately 3 weeks postimplantation by a process of intramembranous bone formation, with the de novo bone tissue observed directly interfacing with the ceramic surface. Infrequently, small isolated areas showed cartilage formation with no noticeable endochondral ossification. At 4 weeks postimplantation of the ceramic with marrow cells, the osteogenesis in the ceramic accompanied an observed increase in compressive strength, rigidity, and energy absorption of the ceramic. These results suggest that a combination of porous ceramics and marrow cells may be useful for clinical problems requiring osseous reconstruction. PMID- 2544712 TI - A study of the mechanical strength of long bone defects treated with various bone autograft substitutes: an experimental investigation in the rabbit. AB - This study was designed to determine which of several bone grafting materials would be the most efficacious substitute for autogenous bone graft in the treatment of segmental long bone defects. The experimental model was a 1-cm defect in the rabbit ulna. The control group had nothing implanted in the defect. The six grafts tested were: (a) autogenous iliac crest bone, (b) autogenous cortical bone (ulna), (c) hydroxylapatite, (d) hydroxylapatite-demineralized bone matrix (allograft) composite graft, (e) freeze-dried bone (allograft), and (f) demineralized bone matrix (allograft). At 6 weeks postoperatively, the ulnas were harvested, examined radiographically, and tested mechanically in torsion. The radiographic examination proved to be of little value because some materials were radiodense at the time of implantation. The rates (percentage) of union, torques at failure, and energy to failure values were statistically significantly higher than control for all groups except hydroxylapatite. We concluded that demineralized bone matrix and hydroxylapatite-demineralized bone matrix composite graft compare favorably with cortical replacement (autograft) in mechanical strength and rate of union and therefore may be satisfactory substitutes for bone grafting. Freeze-dried bone did not appear to be as satisfactory because of its low mean energy to failure, but statistical analysis failed to confirm this opinion. Hydroxylapatite graft, when used alone, does not appear to be a suitable material for grafting segmental bone defects. PMID- 2544713 TI - Effect of intravenously administered immune globulin on functional antibody to herpes simplex virus in low birth weight neonates. PMID- 2544714 TI - Extrarenal Wilms' tumor. AB - The extrarenal location of Wilms' tumor is extremely rare. These tumors can arise from other neoplasms, generally teratomas, or they can present without any associated teratomatous elements. We encountered only 19 well-documented cases of isolated extrarenal nephroblastoma, and we describe three previously unreported patients with this disease. Two of the three children presented also with horseshoe kidneys, an association that may have clinical and embryologic significance. The presence of tumor cephalad to a horseshoe kidney is easier to explain when we consider that they originate from primitive mesodermal tissue, probably mesonephric rests, and not from metanephric remnants. Also, this association should alert physicians to the possible diagnosis of extrarenal nephroblastoma in patients with a retroperitoneal mass and horseshoe kidneys. These patients should be treated according to the same protocols as those for patients with intrarenal Wilms' tumor. PMID- 2544715 TI - Superficial femoral vein transposition in Klippel-Trenaunay syndrome. AB - Use of a superficial femoral vein in competent profunda vein transposition is appropriate in selected cases of Klippel-Trenaunay syndrome; this allows for improvement of venous insufficiency and relief of symptoms secondary to venous stasis. PMID- 2544716 TI - Congenital mesoblastic nephroma, renin production, and hypertension. AB - Between 1964 and 1987, 12 patients with mesoblastic nephroma were treated. The BP was measured preoperatively in five patients, four of whom were hypertensive. Following nephrectomy, the BP returned to normal. Plasma renin levels were measured in one patient; they were markedly elevated pre-operatively, but returned to normal following tumour excision. Immunoreactive renin staining, using a polyclonal antibody to human renin, was performed in the 12 patients. Staining was positive in ten patients. The most intense staining was noted in the areas of cortex entrapped within the tumour, and in perivascular spaces not associated with entrapped cortex. These findings suggest that hypertension secondary to increased tumour associated renin production is a feature of congenital mesoblastic nephroma. PMID- 2544718 TI - Late effects of childhood cancer: a program approach. PMID- 2544717 TI - Comparison of freeze-dried bone allograft and porous hydroxylapatite in human periodontal defects. AB - This study was conducted to clinically compare the efficacy of freeze-dried bone allograft (FDBA) and porous hydroxylapatite granules. Nineteen pairs of intraosseous defects were grafted in seven patients. One defect of each pair was implanted with FDBA, the other with granular porous hydroxylapatite. Matching defects were treated similarly in all other aspects. Evaluations were based on both preoperative and postoperative measurements from a fixed reference point, standardized radiographs, surgical osseous measurements, and histology of degranulated tissues. Grafted sites were reentered 6 to 11 months postsurgery. Results showed a mean osseous fill of 2.1 mm for FDBA versus 1.3 mm for granular porous hydroxylapatite (P = .07). A mean clinical attachment gain of 2.2 mm for FDBA versus 1.3 mm for granular porous hydroxylapatite (P less than .05), and a mean decrease in probing depths of 3.0 mm for FDBA versus 1.4 mm for granular porous hydroxylapatite (P less than 0.5) was found. FDBA was clinically indistinguishable from host bone, whereas porous hydroxylapatite appeared to be separated from host bone by soft tissue. The data and clinical findings suggested that FDBA may have some enhanced reparative potential when compared to granular porous hydroxylapatite in the treatment of periodontal defects in humans. PMID- 2544719 TI - Study of stomach cancer in atomic bomb survivors. Report 1. Histological findings and prognosis. AB - For the purpose of studying the histological findings and prognosis of stomach cancer in atomic bomb survivors, a histological diagnosis of 600 cases of stomach cancer observed between 1964 and 1986 in three medical institutions in Hiroshima City was made by a single pathologist and, furthermore, analysis of 231 cases thereof was performed using the DS86 dose estimated by the Radiation Effects Research Foundation. 1) By histological type of stomach cancer, the frequency of poorly differentiated adenocarcinoma (poorly differentiated adenocarcinoma and signet ring cell carcinoma) was significantly higher in the 1 rad or more group (average stomach organ dose of 30.6 rad) than in the 0 rad group. 2) As for stromal type, the frequency of scirrhous type was significantly higher in the 1 rad or more group, but no difference in infiltrative behavior could be demonstrated. 3) Early cancer tended to be observed at a low rate in the 1 rad or more group. 4) As for average survival time, of the early cancers only well differentiated adenocarcinoma showed a significantly lower survival time in the 1 rad or more group when compared to the 0 rad group. PMID- 2544720 TI - Taste discrimination of traditional and 'health' modified recipes. AB - Beef stew and sponge cake were prepared utilising traditional recipes and recipes modified in accordance with NACNE, National Advisory Committee on Nutrition Education guidelines (1983). Triangle tests were performed to determine whether subjects could discriminate between samples prepared using the traditional recipes. Thirty subjects (15 men and 15 women) participated. No significant differences could be established between the traditional and modified stew or sponge cake in terms of eating quality. Modification of traditional recipes by reducing sugar, fat and salt and increasing the fibre content without affecting the overall sensory quality that the consumer expects is an ideal way of achieving recommended dietary goals. PMID- 2544721 TI - Synthesis of 3-deazaneplanocin A, a powerful inhibitor of S-adenosylhomocysteine hydrolase with potent and selective in vitro and in vivo antiviral activities. AB - The neplanocin A analogue 3-deazaneplanocin A (2b) has been synthesized. A direct SN2 displacement on the cyclopentenyl mesylate 3 by the sodium salt of 6-chloro-3 deazapurine afforded the desired regioisomer 4 as the major product. After deprotection, this material was converted to 3-deazaneplanocin A in two steps. X ray crystallographic analysis confirmed the assigned structure. Consistent with its potent inhibition of S-adenosylhomocysteine hydrolase, 3-deazaneplanocin A displayed excellent antiviral activity in cell culture against vesicular stomatitis, parainfluenza type 3, yellow fever, and vaccinia viruses. Antiviral activity was also displayed in vivo against vaccinia virus by using a mouse tailpox assay. The significantly lower cytotoxicity of 3-deazaneplanocin A, relative to its parent compound neplanocin A, may be due to its lack of conversion to 5'-triphosphate and S-adenosylmethionine metabolites. PMID- 2544722 TI - Inhibition of cyclic adenosine-3',5'-monophosphate phosphodiesterase from vascular smooth muscle by rolipram analogues. AB - Rolipram [(R,S)-4-[3-(cyclopentyloxy)-4-methoxyphenyl]-2-pyrrolidone] has been shown to inhibit selectively the cAMP phosphodiesterase (PDE) of vascular smooth muscle. In order to further explore the structural requirements for selective PDE inhibition, we synthesized a series of rolipram derivatives differently substituted either at the pyrrolidinone or at the aromatic ring. Among these compounds, rolipram was the most active compound. Semirigid analogues were prepared and used for an evaluation of the active conformation of rolipram. Structural comparison with two other potent and chemically different smooth muscle cAMP-PDE inhibitors, trequinsin and Ro 20-1724, allows us to propose a first topological model of the smooth muscle cAMP-PDE pharmacophore. PMID- 2544724 TI - Nonsteroidal cardiotonics. 2. The inotropic activity of linear, tricyclic 5-6-5 fused heterocycles. AB - We previously reported the structure-activity relationships (SAR) of adibendan (1), a potent and long-acting cardiotonic. This paper describes the synthesis of a novel series of linear, tricyclic fused heterocycles of the 5-6-5 type. The compounds were evaluated for positive inotropic activity in anesthetized rats, cats, and dogs. Changes in left ventricular dP/dt were measured as an index of cardiac contractility. The increase in contractility was not mediated via stimulation of beta-adrenergic receptors. The data revealed the intrinsic positive inotropic activity of the parent compound of this series, 5,7-dihydro 7,7-dimethylpyrrolo[2,3-f]benzimidazol-6(1H)-one (2). The structural features that impart optimal inotropic activity are presented and compared with those of the 4,5-dihydro-3(2H)-pyridazinone series. The most potent compounds were evaluated orally in conscious dogs with implanted Konigsberg pressure transducers to measure ventricular pressures, and their effect on left ventricular dP/dt was compared with that of 1, pimobendan, and indolidan. After administration of 1 mg/kg, 1, 3, 7, 19, 22, 24, 31, 54, pimobendan, and indolidan were equipotent, but only with 1, 31, pimobendan, and indolidan, durations of action exceeded 6 h. PMID- 2544723 TI - Synthesis and antiviral activity of the nucleotide analogue (S)-1-[3-hydroxy-2 (phosphonylmethoxy)propyl]cytosine. AB - The acyclic nucleotide analogue (S)-1-[3-hydroxy-2-(phosphonylmethoxy)propyl] cytosine (2, HPMPC) was prepared on a multigram scale in 18% overall yield starting from (R)-2,3-O-isopropylideneglycerol. The key step in the nine-step synthetic route is coupling of cytosine with the side-chain derivative 8 which bears a protected phosphonylmethyl ether group. In vitro data showed that HPMPC has good activity against herpes simplex virus types 1 and 2, although it was 10 fold less potent than acyclovir [AVC, 9-[(2-hydroxyethoxy)methyl]guanine]. By comparison, HPMPC exhibited greater activity than ACV against a thymidine kinase deficient strain of HSV 1 and was more potent than ganciclovir [DHPG, 9-[(1,3 dihydroxy-2-propoxy)methyl]guanine] against human cytomegalovirus. In vivo, HPMPC showed exceptional potency against HSV 1 systemic infection in mice, having an ED50 of 0.1 mg/kg per day (ip) compared with 50 mg/kg per day for ACV. HPMPC was also more efficacious than ACV in the topical treatment of HSV 1 induced cutaneous lesions in guinea pigs. PMID- 2544725 TI - Hexestrol-linked cytotoxic agents: synthesis and binding affinity for estrogen receptors. AB - With the erythro-hexestrol derivative 2 as the starting material, a variety of cytotoxic linked hexestrol (HEX) compounds were prepared including the HEX-N-lost derivative 36, the HEX-(chloroethyl)nitrosourea 38, the HEX-cyclophosphamide 44, and the HEX-epoxide 68. Relative binding affinity to estradiol receptors were in the magnitude of 1%, similar to that of comparable diethylstilbestrol compounds. HEX derivatives with long polyether spacers (64, 65, 70, 71) showed no significant decrease in binding affinity in contrast to derivatives with other bulky side chains. PMID- 2544726 TI - 2(1H)-quinolinones with cardiac stimulant activity. 3. Synthesis and biological properties of 6-imidazol-1-yl derivatives. AB - A series of 6-imidazol-1-yl-8-methyl-2(1H)-quinolinones was synthesized and evaluated for cardiac stimulant activity in dogs. The majority of compounds were prepared from an appropriate 6-imidazol-1-yl-2(1H)-quinolinone precursor or by sulfuric acid catalyzed cyclization of an N-(4-heteroarylphenyl)-3 ethoxypropenamide. Introduction of a range of 5-substituents into 6-(2,4 dimethylimidazol-1-yl)-8-methyl-2(1H)-quinolinone (1) reduced inotropic activity in anesthetized dogs (percentage increase in dP/dtmax) although replacement of the 2-methyl group by iodo (10) or cyano (11) substituents was well tolerated. The 2-methyl-4-chloro (15) and 2-methyl-4-(methylthio) (22) derivatives displayed similar potency to 1 (40-50% increase in dP/dtmax, 10-12.5 micrograms/kg) and these compounds were 3-5 times more potent than milrinone. Introduction of iodo (14), cyano (16), or acetyl (17) substituents into the 4-position approximately halved inotropic activity. In conscious dogs, 11 (0.25 mg/kg) and 16 and 17 (0.125 mg/kg) produced similar increases in cardiac contractility (decrease in the QA interval) to 1 (0.125 mg/kg) and maximum responses were maintained for at least 3 h. Dose-related (25, 125, 250 micrograms/kg) cardiac stimulant activity was demonstrated by 17 and after the higher doses a marked response (approximately 30% increase in dP/dtmax) was still observed after 7 h, in contrast to milrinone. The substantial increases in cardiac contractility observed with 16 and 17 in the conscious dog were not accompanied by any tachycardia. These compounds also displayed an overwhelming selectivity for increasing the force of cardiac contraction (greater than 120% increase in dP/dtmax) rather than heart rate (5-10 beats/min decrease) in the Starling heart lung preparation. As a result of this beneficial pharmacological profile, 6-(4 acetyl-2-methylimidazol-1-yl)-8-methyl-2(1H)-quinolinone (17, UK-66,838) was selected for preclinical development studies. PMID- 2544727 TI - Sodium channel binding and anticonvulsant activities for the enantiomers of a bicyclic 2,4-oxazolidinedione and monocyclic models. AB - Racemic 7-phenyl-9,10-dioxo-1-aza-8-oxabicyclo[5.2.1]decane (1), a bicyclic 2,4 oxazolidinedione that we previously reported was a possible sodium channel anticonvulsant, was resolved into its enantiomeric forms, the absolute configurations were determined, and the stereoisomers were evaluated for relative sodium channel binding and whole animal anticonvulsant activities. Similar studies were carried out with two monocyclic models, 5-ethyl-5-phenyl-2,4 oxazolidinedione (2) and 5-ethyl-3-methyl-5-phenyl-2,4-oxazolidinedione (3). None of these isomers exhibited stereoselective effects in the sodium channel assay, and only modest enantioselectivities were observed for 2 and 3 in the anticonvulsant assays. (R)-(-)-1 was, however, 4 times more toxic than (S)-(+)-1 in the rotorod test, and due to its larger protective index, (S)-(+)-1 exhibited greater therapeutic potential than either (R)-(-)-1 or racemic 1. PMID- 2544728 TI - Exploration of phenyl-spaced 2-amino-(5-9)-phosphonoalkanoic acids as competitive N-methyl-D-aspartic acid antagonists. AB - To investigate the preferred spatial relationship of the distal phosphonic acid to the alpha-amino acid group of the established competitive N-methyl-D-aspartic acid (NMDA) antagonists APH (1) and APV (2), we have prepared a series of ortho-, meta-, and para-substituted (phosphonoalkyl)phenylglycine and -phenylalanine derivatives. With use of a [3H]CPP receptor binding assay, significant binding activity was observed to be critically dependent on both the position of substitution and length of alkyl spacing groups. Two compounds, 4 (phosphonomethyl)-phenylglycine (6, PD 129635) and 3 (phosphonomethyl)phenylalanine (15, PD 130527), displayed receptor-binding affinity comparable to that of APH. Like APH, these compounds were also effective in antagonizing both the proconvulsant and lethal action of NMDA-administered retrobulbar in the mouse. Data are also provided which compare directly the binding efficacy of these compounds against that disclosed recently for the related NMDA antagonist 18 (NPC 451). A preliminary comparison of the structures showing good receptor-binding affinity and in vivo antagonist activity suggests that the NMDA receptor prefers a "folded" rather than "extended" conformation. PMID- 2544729 TI - Angiotensin converting enzyme inhibitors: spirapril and related compounds. AB - The synthesis of spirapril (5), spiraprilat (25), their RSS stereoisomers, and their glycyl (18b) and lysyl (36, 37) analogues is described. These compounds were evaluated in vivo for inhibition of angiotensin converting enzyme (ACE), and selected compounds were evaluated for in vitro ACE inhibition (spirapril ID50 16 micrograms/kg; spiraprilat IC50 0.8 nM, ID50 8 micrograms/kg). In anesthetized rats, iv, esters 5 and 36 are more potent than enalapril, and diacids 25 and 37 are more potent than enalaprilat in vitro. In the conscious rats, orally, 5 and enalapril (2) showed potent and sustained activity at doses of 0.03-1 and 0.1-1 mg/kg, respectively. From this work, spirapril was selected for clinical evaluation as an antihypertensive agent. PMID- 2544730 TI - Nucleotide sequences of Caenorhabditis elegans core histone genes. Genes for different histone classes share common flanking sequence elements. AB - We have determined the nucleotide sequence of core histone genes and flanking regions from two of approximately 11 different genomic histone clusters of the nematode Caenorhabditis elegans. Four histone genes from one cluster (H3, H4, H2B, H2A) and two histone genes from another (H4 and H2A) were analyzed. The predicted amino acid sequences of the two H4 and H2A proteins from the two clusters are identical, whereas the nucleotide sequences of the genes have diverged 9% (H2A) and 12% (H4). Flanking sequences, which are mostly not similar, were compared to identify putative regulatory elements. A conserved sequence of 34 base-pairs is present 19 to 42 nucleotides 3' of the termination codon of all the genes. Within the conserved sequence is a 16-base dyad sequence homologous to the one typically found at the 3' end of histone genes from higher eukaryotes. The C. elegans core histone genes are organized as divergently transcribed pairs of H3-H4 and H2A-H2B and contain 5' conserved sequence elements in the shared spacer regions. One of the sequence elements, 5' CTCCNCCTNCCCACCNCANA 3', is located immediately upstream from the canonical TATA homology of each gene. Another sequence element, 5' CTGCGGGGACACATNT 3', is present in the spacer of each heterotypic pair. These two 5' conserved sequences are not present in the promoter region of histone genes from other organisms, where 5' conserved sequences are usually different for each histone class. They are also not found in non-histone genes of C. elegans. These putative regulatory sequences of C. elegans core histone genes are similar to the regulatory elements of both higher and lower eukaryotes. The coding regions of the genes and the 3' regulatory sequences are similar to those of higher eukaryotes, whereas the presence of common 5' sequence elements upstream from genes of different histone classes is similar to histone promoter elements in yeast. PMID- 2544731 TI - Enhancer binding protein (EBP1) makes base and backbone contacts over one complete turn of the DNA double helix. AB - The simian virus 40 (SV40) enhancer consists of multiple DNA sequence motifs that represent the binding sites for a large number of trans-acting factors. We have purified one such factor, EBP1, which binds to a region encompassing the "core" of the SV40 enhancer, and appears to be involved in transcriptional activation. The interaction of EBP1 with its recognition site has been analysed by nuclease protection and by a variety of chemical probes. Enhancer sequences protected from cleavage with DNase I in the presence of EBP1 extend from position 232 to 250 on one strand and from 233 to 251 on the other strand. Methylation protection and alkylation interference studies have identified purine bases and backbone phosphate groups that participate in the formation of a specific EBP1-DNA complex. Within a ten base-pair region, every purine base interferes with binding when methylated and six phosphate groups on each strand interfere with binding when the attached oxygen groups are ethylated. "Footprinting" with hydroxyl radicals, generated by the 1,10-orthophenanthroline-copper ion, revealed sugar residues in the binding site that were protected from cleavage in the presence of EBP1. Computer graphics analyses of the contact point data indicate that EBP1 makes base and backbone contacts with the DNA over one complete turn of the DNA double helix, and suggest a model in which EBP1 makes sequence-specific contacts in the major groove, although binding may be influenced by interactions in the minor groove. Comparison of the EBP1 contact points with that of other known DNA binding proteins indicates that EBP1 employs a unique mechanism to recognize a specific DNA sequence. PMID- 2544732 TI - Crystallization of yeast iso-2-cytochrome c using a novel hair seeding technique. AB - A hair seeding technique has been developed to obtain diffraction quality crystals of yeast (Saccharomyces cerevisiae) iso-2-cytochrome c, a model for studies of protein folding and biological electron transfer reactions. Deep red crystals of this protein were obtained from 88 to 92% saturated solutions of ammonium sulfate containing 20 mg protein/ml, 0.1 M-sodium phoshate, 0.3 M-sodium chloride, 0.04 M-dithiothreitol and adjusted to phosphate, 0.3 M-sodium chloride, 0.04 M-dithiothreitol and adjusted to pH 6.0. Rapid crystal growth was observed, but only along the path of the seeding hair stroke. The space group is P4(3)2(1)2 (or P4(1)2(1)2) with a = b = 36.4 A, c = 137.8 A (1 A = 0.1 nm) and Z = 8. Crystals are stable in the X-ray beam for more than 10 days and diffract to at least 2.5 A resolution. The same hair seeding methodology has proven useful in obtaining crystals of specifically designed mutant iso-2 proteins and in other protein systems where consistent crystal growth had previously proven difficult to attain. PMID- 2544733 TI - Adenylate kinases from thermosensitive Escherichia coli strains. AB - The adk genes from several thermosensitive (ts) mutants of Escherichia coli were cloned and sequenced. The mutations responsible for the thermolability of the gene product, the enzyme adenylate kinase, were established. From five independently isolated strains analysed, two contain a CCG to TCG transition changing proline 87 to serine (P87S), another two have a TCT to TTT transition that mutates serine 129 to phenylalanine (S129F), and the last one was found not to contain a mutation in the adk gene. Overproducing strains were constructed that contain ts genes in the genome as well as in the plasmids. These strains grow at high temperature, although much slower than wild-type. Most probably, the high rate of synthesis of adenylate kinase compensates for the destruction of the thermolabile protein by the elevated temperature. Mutated proteins were purified. The P87S but not the S129F mutation was found to cause thermosensitivity of the adenylate kinase reaction. Revertants of thermosensitivity were isolated and the nature of the mutation was determined by the RNase digestion method of RNA-DNA hybrids and by DNA sequencing. The revertants of the P87S mutation regained the wild-type sequence, whereas the revertants of the S129F strain retained the original mutation in the adenylate kinase gene. These results are discussed in the light of the three-dimensional structure of the enzyme and the possible role of adenylate kinase in phospholipid synthesis. PMID- 2544734 TI - Three-dimensional structures of drug-resistant mutants of human rhinovirus 14. AB - Mutants of human rhinovirus 14 were isolated and characterized by searching for resistance to compounds that inhibit viral uncoating. The portions of the RNA that code for amino acids that surround the antiviral compound binding site were sequenced. X-ray analysis of two of these mutants, 1188 Val----Leu and 1199 Cys-- -Tyr, shows that these were single-site substitutions which would sterically hinder drug binding. Differences in the resistance of mutant viruses to various antiviral compounds may be rationalized in terms of the three-dimensional structures of these mutants. Predictions of the structures of mutant rhinovirus 14 with the substitutions 1188 Val----Leu, 1199 Cys----Tyr and 1199 Cys----Trp in VP1 were made using a molecular dynamics technique. The predicted structure of the 1199 Cys----Tyr mutant was consistent with the electron density map, while the 1188 Val----Leu prediction was not. Large (up to 1.4 A) conformational differences between native rhinovirus 14 and the 1199 Cys----Tyr mutant occurred in main-chain atoms near the mutation site. These changes, as well as the orientation of the 1199 tyrosine side-chain, were correctly predicted by the molecular dynamics calculation. The structure of the predicted 1199 Cys----Trp mutation is consistent with the drug-resistant properties of this virus. PMID- 2544735 TI - Strong inclination toward transition mutation in nucleotide substitutions by poliovirus replicase. AB - A viable insertion mutant of the Sabin strain of type 1 poliovirus was constructed. The mutant carried an insertion sequence of 72 nucleotides at nucleotide position 702 in the 5' non-coding region (742 nucleotides long) of the genome of the Sabin strain. This mutant showed a small-plaque phenotype, as compared with the parental virus. Indeed, the final yield of the mutant in a single cycle of infection was tenfold fewer than that of the parental virus. Many large-plaque variants that are easily generated from the insertion mutant appeared to regain efficient viral replication and have single nucleotide changes. All nucleotide changes observed were limited to within three nucleotides of an AUG sequence in the insertion sequence. The result indicates strongly that the AUG sequence itself in this genome region functions in reducing the plaque size of the parental Sabin type 1 virus. The insertion mutant with a small-plaque phenotype may be the first in vitro mutant of poliovirus whose viability is lowered only by a primary sequence inserted into the 5' non-coding region of the genome. Base substitutions to alter the AUG sequence should largely be the result of errors of the virus-specific replicase, since variants with base substitutions must be subject to only minimum selection pressure. Accordingly, nucleotide sequence analysis of the genome region containing the AUG sequence was performed on a number of genomes of large-plaque variants to investigate types of nucleotide substitutions caused by characteristic errors in RNA replication. Only one transversion mutation was detected in the genomes of 44 independently isolated large-plaque variants with single base changes in the AUG sequence. This result suggests strongly that transition mutations occur predominantly as nucleotide substitutions caused by characteristic errors of poliovirus replicase. PMID- 2544736 TI - Effect of nucleosome distortion on the linking deficiency in relaxed minichromosomes. AB - The wrapping of closed circular DNA on a protein surface, followed by relaxation with a topoisomerase and removal of proteins, produces a characteristic DNA linking deficiency, delta Lk. We show that the magnitude of delta Lk depends upon the surface shape, and we calculate changes in delta Lk caused by particular distortions of the protein wrapping surface. If the DNA remains attached to the surface during distortion, the DNA winding number, phi, is not altered. The change in delta Lk is then equal to the change in the surface linking number, SLk, which is a straightforward measure of the wrapping of the DNA around the surface. For left-handed wrapping, as in a nucleosome, SLk = -n, the number of times that the DNA axis winds around the axis of the protein complex. We calculate values of SLk for the helical wrapping of a constant length of DNA on protein surfaces having the shapes of cylinders and of ellipsoids and hyperboloids of revolution. If the equatorial radius of the protein is fixed, change in shape from a cylinder to a hyperboloid increases SLk, while the corresponding change to an ellipsoid reduces SLk. We apply the general results to the interpretation of experiments in which minichromosomes are relaxed with topoisomerase at various temperatures and delta Lk is determined. The result is that a distortion of the nucleosome core by at most 5% (the change in the radius at the axial extremity relative to the equator) is sufficient to explain the observed delta Lk changes. PMID- 2544738 TI - Transposon Tn7. cis-Acting sequences in transposition and transposition immunity. AB - We have identified and characterized the cis-acting sequences at the termini of the bacterial transposon Tn7 that are necessary for its transposition. Tn7 participates in two kinds of transposition event: high-frequency transposition to a specific target site (attTn7) and low-frequency transposition to apparently random target sites. Our analyses suggest that the same sequences at the Tn7 ends are required for both transposition events. These sequences differ in length and nucleotide structure: about 150 base-pairs at the left end (Tn7L) and about 70 base-pairs at the right end (Tn7R) are necessary for efficient transposition. We also show that the ends of Tn7 are functionally distinct: a miniTn7 element containing two Tn7R ends is active in transposition but an element containing two Tn7L ends is not. We also report that the presence of Tn7's cis-acting transposition sequences anywhere in a target replicon inhibits subsequent insertion of another copy of Tn7 into either an attTn7 target site or into random target sites. The inhibition to an attTn7 target site is most pronounced when the Tn7 ends are immediately adjacent to attTn7. We also show that the presence of Tn7R's cis-acting transposition sequences in a target replicon is necessary and sufficient to inhibit subsequent Tn7 insertion into the target replicon. PMID- 2544737 TI - Ligand and proton exchange dynamics in recombinant human myoglobin mutants. AB - Site-specific mutants of human myoglobin have been prepared in which lysine 45 is replaced by arginine (K45R) and aspartate 60 by glutamate (D60E), in order to examine the influence of these residues and their interaction on the dynamics of the protein. These proteins were studied by a variety of methods, including one and two-dimensional proton nuclear magnetic resonance spectroscopy, exchange kinetics for the distal and proximal histidine NH protons as a function of pH in the met cyano forms, flash photolysis of the CO forms, and ligand replacement kinetics. The electronic absorption and proton nuclear magnetic resonance spectra of the CO forms of these proteins are virtually identical, indicating that the structure of the heme pocket is unaltered by these mutations. There are, however, substantial changes in the dynamics of both CO binding and proton exchange for the mutant K45R, whereas the mutant D60E exhibits behavior indistinguishable from the reference human myoglobin. K45R has a faster CO bimolecular recombination rate and slower CO off-rate relative to the reference. The kinetics for CO binding are independent of pH (6.5 to 10) as well as ionic strength (0 to 1 M NaCl). The exchange rate for the distal histidine NH is substantially lower for K45R than the reference, whereas the proximal histidine NH exchange rate is unaltered. The exchange behavior of the human proteins is similar to that reported for a comparison of the exchange rates for myoglobins having lysine at position 45 with sperm whale myoglobin, which has arginine at this position. This indicates that the differences in exchange rates reflects largely the Lys----Arg substitution. The lack of a simple correlation for the CO kinetics with this substitution means that these are sensitive to other factors as well. Specific kinetic models, whereby substitution of arginine for lysine at position 45 can affect ligand binding dynamics, are outlined. These experiments demonstrate that a relatively conservative change of a surface residue can substantially perturb ligand and proton exchange dynamics in a manner that is not readily predicted from the static structures. PMID- 2544739 TI - Genetic analysis of attTn7, the transposon Tn7 attachment site in Escherichia coli, using a novel M13-based transduction assay. AB - The large (14 kb; kb = 10(3) bases) bacterial transposon, Tn7 (encoding resistance to trimethoprim and streptomycin/spectinomycin), has unusual properties. Like other elements, Tn7 transposes with low efficiency and low target-site specificity, but Tn7 also transposes, with high frequency in a unique orientation, to a preferred "attachment" site, called attTn7, in the Escherichia coli chromosome and similarly into plasmids containing attTn7. We developed a novel bacteriophage M13-based assay system to measure the transposition frequency of Tn7 to M13mp phage vectors containing attTn7 on a cloned 1 kb fragment of chromosomal DNA. Phage harvested from a Tn7 donor strain were used to infect recipient bacteria with selection for trimethoprim resistance. Transposition frequency, expressed as the number of trimethoprim-resistant colonies per plaque forming unit, was found to be approximately 10(-4) to M13mp::attTn7, in contrast to 10(-10) to M13mp recombinants with approximately 1 kb insertions of other, "generic brand", DNA. By deletion analysis of M13mp::attTn7, we show that attTn7 is contained within a 64 base-pair region; sequences adjacent to the actual insertion site and encoding the carboxy terminus of the glmS gene are required. This assay also provided evidence for transposition immunity conferred by the right end of Tn7. PMID- 2544740 TI - Influence of docosahexaenoic acid on cisplatin resistance in a human small cell lung carcinoma cell line. AB - In a sensitive, human, small cell lung carcinoma cell line (GLC4) and a cisplatin (CP)-resistant subline (GLC4-CP), the effect of co-culturing with docosahexaenoic acid (DCHA) on CP cytotoxicity was studied. Cells were cultured for 4 days, with 32 microM of DCHA added on days 1 and 3. Incorporation of DCHA into the cellular phospholipids was demonstrated by fatty acid analysis. Supplementation with DCHA led to almost a threefold decrease of resistance in GLC4-CP and had no influence on CP cytotoxicity in GLC4. After culturing with DCHA, cellular platinum (Pt); total Pt bound to DNA; and Pt-GG, Pt-AG, G-Pt-G, and Pt-GMP adduct contents increased in both lines, whereas interstrand cross-link formation was elevated only in GLC4-CP. These experiments demonstrate that DCHA reduces CP resistance. Although an effect on cellular membranes resulting in an increased CP uptake apparently was present, this mechanism does not seem to be responsible for resistance modulation. Rather, an effect on nuclear, probably DNA-related, structures is likely and leads to an increased formation of interstrand cross links in GLC4-CP. PMID- 2544741 TI - Loss of leukoregulin up-regulation of natural killer but not lymphokine-activated killer lymphocytotoxicity in human papillomavirus 16 DNA-immortalized cervical epithelial cells. AB - The sensitivity of human cervical epithelial cells immortalized by transfection with human papillomavirus type 16 (HPV16) DNA, to lysis by natural killer (NK) and lymphokine-activated killer (LAK) lymphocytes was evaluated at progressive stages of transformation. Both early- (10-20 wk) and late- (greater than 30 wk) passage HPV16-immortalized cells were resistant to NK lymphocyte cytotoxicity but sensitive to LAK lymphocyte cytotoxicity at lymphocyte-to-cervical cell ratios ranging from 1:1 to 50:1 in a 4-hour 51Cr release assay. Treatment of early passage HPV16 DNA-immortalized cells with 2.5 U/mL of the NK lymphocytotoxicity sensitizing lymphokine, leukoregulin, for 1 hour induced modest sensitivity to NK cells (P less than .05) but markedly up-regulated LAK sensitivity twofold to threefold. At the later passages, leukoregulin up-regulation of sensitivity to NK was lost but remained to LAK lymphocytotoxicity. Similarly, an HPV16-positive human cervical carcinoma cell line, QGU, was also resistant to NK lymphocytotoxicity and sensitive to LAK lymphocytotoxicity; leukoregulin failed to confer sensitivity to the NK-resistant QGU tumor cells and increased their sensitivity to LAK lymphocytotoxicity 1.5-fold to twofold. Although the HPV immortalized cervical cells containing integrated HPV16 DNA were not tumorigenic, they mimicked the response of established HPV16-positive cervical carcinoma cells. HPV16-immortalized cervical epithelial cells provide a useful model for the study of cytokine modulation of dysplastic and neoplastic cervical epithelial cell sensitivity to natural lymphocytotoxicity. PMID- 2544742 TI - A prospective double-blind trial of a low molecular weight heparin once daily compared with conventional low-dose heparin three times daily to prevent pulmonary embolism and venous thrombosis in patients with hip fracture. AB - In a randomized, prospective, double-blind trial, the effect of conventional low dose heparin (5,000 units every 8 hours) was compared with that of a low molecular weight fragment (2165 LMWH, Kabi Vitrum AB, starting 2,500 units 2 hr before surgery, and then 5,000 units subcutaneously every morning for 9 days). A total of 90 patients admitted because of hip fracture fulfilled the inclusion criteria and were analyzed for development of pulmonary embolism and deep vein thrombosis: 46 patients were included in the low molecular weight heparin (LMWH) group, and 44 in the conventional heparin group. Two and three, respectively, died before diagnostic tests were performed. In the remaining patients a ventilation-perfusion lung scan was performed 8 days after intervention. In the first 57 patients studied a bilateral ascending venography was performed on the ninth day only if clinical symptoms suggested a deep venous thrombosis. Because of the rate of venous thrombosis detected in those patients was unexpectedly low, venography was requested in the remaining 33 patients, even if the screening tests were negative. Pulmonary embolism occurred in six patients, all in the LMWH group. Deep vein thrombosis occurred in 14 patients in the LMWH group and in six patients in the conventional heparin group. Both differences are statistically significant. Mortality did not differ between the groups, nor did haemorrhagic complications. Our findings suggest that, in patients with hip fracture, LMWH is not useful at the dosage used. PMID- 2544743 TI - Diseases that mimic herpes simplex encephalitis. Diagnosis, presentation, and outcome. NIAD Collaborative Antiviral Study Group. AB - A total of 432 patients underwent brain biopsy for presumptive herpes simplex encephalitis. Three patient groups were identified. The first group, 195 patients (45%), had herpes simplex encephalitis confirmed by the isolation of herpes simplex virus from brain tissue at biopsy (193 patients) or autopsy (2 patients). The second group, 95 patients (22%), had diseases that were identified but that were not caused by herpes simplex virus. Three subgroups were recognized: (1) 38 patients (9%) with treatable disease, (2) 40 patients (9%) with nontreatable but diagnosed viral infection, and (3) 17 patients (4%) with identified diseases neither of viral etiology nor treatable. The third group, 142 patients (33%), remained without a diagnosis. Clinical presentation of patients in the second group was similar to that of those with herpes simplex encephalitis and those without a diagnosis. Patients in the subgroup with nontreatable but diagnosed viral infections had the greatest likelihood of returning to normal. PMID- 2544744 TI - From the Centers for Disease Control. General recommendations on immunization. PMID- 2544745 TI - CDC's approach to firearm injuries. PMID- 2544746 TI - Dietary fiber and health. Council on Scientific Affairs. AB - During the last 18 years, considerable research has been conducted on the role of dietary fiber in health and disease. Interest was stimulated by epidemiologic studies that associated a low intake of dietary fiber with the incidence of colon cancer, heart disease, diabetes, and other diseases and disorders. Dietary fiber is not a single substance. There are significant differences in the physiological effects of the various components of dietary fiber. A Recommended Dietary Allowance for dietary fiber has not been established. However, an adequate amount of dietary fiber can be obtained by choosing several servings daily from a variety of fiber-rich foods such as whole-grain breads and cereals, fruits, vegetables, legumes, and nuts. PMID- 2544747 TI - [Clinical studies on the relation of abnormal growth hormone secretions to hepatic diabetes mellitus in patients with hepatocellular carcinoma]. AB - Paradoxical growth hormone (GH) responses in 50 g or 75 g oral glucose tolerance tests (OGTT) have been demonstrated in 24 patients with hepatocellular carcinoma, whereas no significant changes in serum GH levels after OGTT were shown in 10 normal controls, 6 patients with cirrhosis of liver, and with chronic active hepatitis. There were no significant difference in the GH responses in OGTT as well as in the incidence of paradoxical GH responses between diabetic and non diabetic patients with HCC. Informatively, the basal somatomedin C level was very low in all cases examined. PMID- 2544748 TI - [Clinical analysis of patients with hepatocellular carcinoma with non-A, non-B hepatitis virus]. AB - An analysis of 242 patients with a hepatocellular carcinoma (HCC) has shown that 65 patients without a hepatitis B virus (HBV), alcohol abuse, or a transfusion history and 17 posttransfusion patients had a mean death age of 65.5 +/- 10.2 and 62.1 +/- 8.4 years respectively. The average age of 65 patients was 8.5 years older than those with a HBV and without a alcohol abuse history. Of 17 patients, 3 with a HBV had a mean death age of 55.3 +/- 10.8 years and the interval time from transfusion to a HCC was significantly shorter than in other post transfusion patients. PMID- 2544750 TI - [A histopathologic comparative study of European cases of fibrolamellar carcinoma and Japanese cases of sclerosing hepatocellular carcinoma]. AB - Ten fibrolamellar carcinomas (FLCs), 3 FLC-like cancers, and 5 cases of a sclerosing hepatocellular carcinoma (SHCC) were examined clinicopathologically. The FLCs were found in young people in an equal sex ratio and showed no indications of an HBsAg, AFP, or cirrhosis. FLC cancer cell showed a large eosinophilic cytoplasm and prominent nucleoli. SHCCs were found in middle-aged people, male-predominant, and often positive for HBsAg and AFP. Cancer cells of the SHCC type showed less eosinophilic small cytoplasms and inconspicuous nucleoli. Although these two types are fairly distinctive entities, there were a few still unclassified HCCs with fibrous stroma. Further precise criteria would seem to be required for an accurate diagnosis of FLC. PMID- 2544749 TI - [Influence of hepatic ischemia on liver regeneration following hepatectomy, with special reference to the therapeutic choice for ruptured hepatoma]. AB - The influence of partial hepatic ischemia (32%) prior to partial hepatectomy (68%) has been studied in the rat. 3H-thymidine incorporation into the hepatic DNA was significantly suppressed in both 20 min and 30 min ischemia depressed the survival following partial hepatectomy (p less than 0.001). Three cases of ruptured hepatocellular carcinomas were treated: one case by emergency hepatectomy, and two cases by hepatectomy following TAE. Hepatic insufficiency and post-operative death occurred to only the case given an emergency hepatectomy. Thus, it is felt that a ruptured hepatoma should first be treated by TAE and then surgically resected. PMID- 2544752 TI - [Papilloma virus infection of mothers and infants]. PMID- 2544751 TI - [Determination of cadmium in urine by flameless atomic absorption spectrophotometry]. PMID- 2544754 TI - Human herpesvirus Epstein-Bar virus: ubiquity, oncogenicity, and risk factors. PMID- 2544753 TI - Viral myocarditis and cardiomyopathy. PMID- 2544755 TI - Morning and evening adrenocortical responses to ACTH stimulation in endogenously depressed patients: a preliminary report. AB - Adrenocortical stimulation with ACTH both in the morning (M-test) and in the evening (E-test) and the dexamethasone suppression test were carried out in patients suffering from endogenous depression (DEP) and normal controls (NOR). A greater cortisol release in DEP was recognized than in NOR in the M-test, an earlier peak response of DEP was shown in the M-test than in the E-test, and a lack of association between hypersecretion of cortisol during depression and cortisol output after ACTH administration was noted. These findings, together with the results of DST, suggest that excessive activity of the hypothalamic pituitary-adrenal (HPA) axis in depression may result, partly, from adrenocortical hyperresponsiveness. PMID- 2544756 TI - [A case of bilateral multiple lung cancer resected by synchronous bilateral thoracotomy]. AB - A 59-year-old female with multiple lung cancer, located in left upper lobe and in right upper lobe received synchronous bilateral thoracotomy. The patient underwent left upper division segmentectomy and right S3 segmentectomy. Histological examination showed large cell carcinoma in the left side and small cell carcinoma in the right side. The patient died 5 years and 3 months after operation. If general condition and pulmonary function permit, bilateral thoracotomy for multiple lung cancer is recommended. PMID- 2544757 TI - [A case of the successful surgical treatment of mediastinal giant neurogenic tumor, and the chest wall reconstruction by the autologous ribs]. AB - A mediastinal tumor, particularly a neurogenic tumor, sometimes develops into a giant tumor; this large tumor mass then puts pressure on the heart and lungs, and may cause severe complications. In our department, 164 mediastinal tumors were experienced, the largest one was an intrathoracic fibromyoma which weighed 3,760 g. This report deals with our recent experience in which we found a neurogenic tumor weighed out 4,500 g. This tumor was excised in an emergency operation, after which the patient's life was saved by twenty-two days of artificial respiration. Since this tumor was so large, four entire right ribs had to be removed in order to create enough space to excise it. In order to reconstruct the chest wall, three of the removed ribs were retransplanted in their original sites. An extremely good prognosis resulted. PMID- 2544758 TI - Comparison of feline parvovirus subspecific strains using monoclonal antibodies against a feline panleukopenia virus. AB - Four monoclonal antibodies (mAb) against a feline panleukopenia virus (FPLV) TU 1 strain, one of the host range variants of feline parvovirus (FPV), were produced and applied for antigenic analysis of FPLV, canine parvovirus (CPV) and mink enteritis virus (MEV). All mAbs were considered to be directed at epitopes on the virus capsid surface because they neutralized the infectivity and inhibited the hemagglutination (HA) of the homologous virus as well as other FPV strains. They were of the mouse IgG1 type. High antigenic homogeneity among FPLV strains was confirmed by HA-inhibition (HI) test with the mAbs and polyclonal immune sera against FPLV or CPV. But the TU 11 strain of FPLV was antigenically distinguished from the remaining 14 FPLV strains by both the HI test and the micro neutralization test with one of the mAbs produced. MEV Abashiri strain was found to be antigenically indistinguishable from FPLV. Most of the CPV strains isolated after 1981 were considered to be antigenically different from earlier CPV isolates when some mAbs were applied in the serological tests, confirming the replacement of CPV by an antigenic variant in Japan. However, antigenically different CPVs were detected at the end of 1984 from unrelated epizootics occurred a month apart in the same area. PMID- 2544759 TI - Protection against bovine rotaviruses in newborn calves by continuous feeding of immune colostrum. AB - Three pregnant cows were inoculated intramuscularly with inactivated vaccine to bovine rotavirus (BRV) serotype 1 (BRV-1) and serotype 2 (BRV-2). Serum neutralizing antibody (NA) titers against both serotypes increased significantly after immunization. NA titers of colostrum obtained from immunized cows against BRV-1 and BRV-2 were 29286 and 38109, respectively, which were significantly higher than those from non-immunized control cows. Nine and 6 colostrum deprived calves were orally challenged with BRV-1 and BRV-2, respectively, and monitored for clinical manifestation and viral shedding. Five calves of them, 3 with BRV-1 and 2 with BRV-2, received 2 l of milk replacer supplemented with 10% immune colostrum 2 hr before challenge and twice daily for the first 5 days after challenge. Other 10 calves, 6 with BRV-1 and 4 with BRV-2, were fed only milk replacer as controls. All control calves developed severe diarrhea and shed a large amount of BRV in feces, beginning from 24 to 48 hr after challenge inoculation. On the contrary, all calves but one fed colostrum supplement remained clinically healthy after challenge, and BRV was not detected in their feces during feeding immune colostrum. The possibility that continuous feeding of immune colostrum is capable of preventing newborn calves from diarrhea associated with BRV and viral shedding was suggested. PMID- 2544760 TI - Characterization of a parvovirus isolated from the diarrheic feces of a pig. AB - A small DNA virus was isolated from the feces of a sow with diarrhea and identified as a parvovirus on the basis of its properties. The virus replicated preferentially in cell cultures of swine origin, including primary porcine thyroid gland and kidney cell cultures in which the cytopathic effect developed. The virus agglutinated erythrocytes of guinea pig, mouse and human group O but not these of chicken. The growth of the virus was inhibited by 5-iodo-2' deoxyuridine. The virus was resistant to ether and heating at 56 degrees C for 30 min and stable at pH 3.0. The buoyant density of the infectious particles was 1.40 g/ml in CsCl density gradient, and the virions were 27 nm in diameter by electron microscopy. The viral protein seemed to be separated into four polypeptides with molecular weights of 81k, 70k, 66k and 62k daltons respectively. Cross serum neutralization test demonstrated that the virus was antigenically different from porcine parvovirus as well as bovine and canine parvoviruses. These findings and the survey on neutralizing antibody distribution indicated indirectly that another parvovirus which could be antigenically distinguished from well-known porcine parvovirus had been widespread among swine in Japan. PMID- 2544761 TI - Effect of dietary sodium bicarbonate and magnesium oxide on cows with milk fat depression in several dairy herds. AB - Buffer mainly consisting of 100 g of sodium bicarbonate and 30 g of magnesium oxide was added to the feed per head per day and given for 8 months to groups of 92 cows of milk fat depression. Milk fat increased from 3.06% (pre-treatment) to 3.68% at 4 months and 3.71% at 8 months post-treatment. Solids-not-fat was slightly increased by the buffers. Milk production was not affected. The number of rumen protozoa increased from 2.85 X 10(5)/ml (pre-treatment) to 9.61 X 10(5)/ml at 8 months post-treatment and the kinds of protozoa diversified. An increase of acetate and decrease of propionate were observed together with increased milk fat at 8 months post-treatment. An increase of hematocrit, lactate dehydrogenase, sodium and potassium, and a decrease of cholesterol and gamma glutamyl transpeptidase in blood were recognized after the treatment. The incidence of disease was reduced. There was a significant correlation between increased milk fat percentage and increase in the number and the kinds of protozoa. PMID- 2544762 TI - Changes in serum isozymes in breeding Japanese black heifer with Ibaraki disease. PMID- 2544763 TI - [Vascular purpura and other cardiovascular alterations in children with acute parvovirus B19 infection. Report of 3 personal cases]. AB - Most frequent skin manifestation of parvovirus B19 infection is the erythema infectiosum (fifth disease; Sticker's disease). Vascular purpura (some cases of Henoch-Schonlein purpura) has been reported in virus infections such as infectious mononucleosis (Epstein-Barr virus), hepatitis B, cytomegalovirus etc., and human parvovirus B19 could be another cause. Clinical course, procedure of diagnostics and therapy in 3 own serologically established cases of vascular purpura or severe alterations of the blood circulation due to parvovirus B19 infection are presented. Parvovirus B19 (or other viruses) as a trigger for vascular purpura and alteration of the blood vessel wall and blood circulation could explain the childhood predominance and seasonal clustering of that disease. Furthermore an individual disposion (in form of special HLA type?) for the development of vascular purpura can be suggested. PMID- 2544764 TI - [Reoperations in adenomas of the bronchus]. AB - The authors discuss their experience in the treatment of 18 patients who were operated on for a second time due to the development of a recurrent tumor. A true and false recurrence was distinguished. The causes of their development were: diagnostic errors in 6, malignancy of the adenoma in 4, and technical faults during the operation in 8 patients. Various types of repeated interventions were conducted in in all 18 patients, preference was given to reconstructive-plastic operations. There were no fatal outcomes. Fourteen patients have a survival period of 5 months to 17 years, four patients with malignant carcinoid died from metastases. PMID- 2544765 TI - [Treatment of patients with cancer of the colon]. AB - Experience in the treatment of 317 patients with cancer of the colon is discussed. Surgery was conducted in 226 patients, 91 patients received combined treatment (operation and intravenous injection of radioactive colloidal gold). Postoperative mortality was 5.99%. The main cause of death was peritonitis (4.1%) due to incompetence of the anastomosis. Combined treatment of patients with stage III cancer of the colon improves somewhat the late-term results of management. PMID- 2544766 TI - [Use of emergency cytological studies in bronchoscopic diagnosis of pulmonary cancer]. AB - The article deals with the indications for emergency cytological study of central and peripheral cancer of the lung which were determined on the basis of examination of 268 patients. The significance of this method in detecting pulmonary carcinoma and reducing the number of repeated bronchoscopies is shown. A method for quick staining of the specimens is suggested and the role of emergency cytological study in complex examination of patients with cancer of the lung is shown. Emergency cytological study of material taken during bronchoscopy made it possible to verify the diagnosis in 77.7% of patients. PMID- 2544767 TI - [Combined treatment of soft tissue sarcoma]. AB - The five-year results of surgical and combined (irradiation + operation) treatment of 451 patients with sarcoma of the soft tissues are analysed with due account for the principal prognostic factors. It is shown that the survival of patients may be prolonged if the operation is supplemented by irradiation. Comparison of three variants of combined treatment showed the advantages of combined treatment with subsequent postoperative irradiation. Radiotherapy does not increase the number of postoperative complications. PMID- 2544768 TI - [Aspiration puncture biopsy in the preoperative diagnosis of adrenal tumors]. AB - The results of fine-needle aspiration puncture (AP) and cytologic examination in the preoperative period in 4 patients with various lesions of the adrenals are discussed. AP was conducted under local anesthesia and control by ultrasonic examination; no complications occurred. Analysis of the cytologic findings allowed preoperative morphological verification of the process, as a result of which the initial clinical diagnosis was changed in 2 of the 4 cases. With an exactly established preoperative cytologic diagnosis a rational therapeutic tactics can be chosen, and the necessity of using complicated examination methods, diagnostic laparotomy among others, obviated. PMID- 2544769 TI - Epidermal growth factor receptors and epidermal growth factor-like activity in colorectal mucosa, adenomas and carcinomas. AB - The epidermal growth factor (EGF) and alpha-tumor growth factor are mitogenic proteins which bind to the EGF-receptor and may play a role in carcinogenesis or tumor progression. Our study investigated whether colorectal carcinomas and adenomas express altered levels of EGF-receptors or overproduce EGF-like activity by comparing histologically normal mucosa to carcinomas resected from the same patients. EGF-receptors were characterized by radioligand binding studies. Carcinomas contained unchanged or decreased levels of EGF-receptors in 13/16 and moderately increased levels in 3/16 patients as compared to normal mucosa. Adenomas obtained from 2 patients with familial polyposis coli and from a third patient with a coincident carcinoma had similar numbers of EGF-receptors as normal mucosa. EGF-like growth factors, in contrast, were significantly elevated in carcinoma extracts as compared to extracts from normal mucosa of the same patients. Adenomas did not contain elevated levels of EGF-like activity. We conclude that increased expression of EGF-receptors is infrequent in colonic adenocarcinomas. Increased production of EGF-like growth factors may frequently occur but seems to be associated with tumor progression rather than with premalignant lesions as represented by adenomas. PMID- 2544770 TI - Second messenger systems and progesterone secretion in the small cells of the bovine corpus luteum: effects of gonadotropins and prostaglandin F2a. AB - The present studies were conducted to determine the effects of gonadotropins (LH and hCG) and prostaglandin F2a (PGF2a) on the production of "second messengers" and progesterone synthesis in purified preparations of bovine small luteal cells. Corpora lutea were removed from heifers during the luteal phase of the normal estrous cycle. Small luteal cells were isolated by unit-gravity sedimentation and were 95-99% pure. LH provoked rapid and sustained increases in the levels of [3H]inositol mono-, bis-, and trisphosphates (IP, IP2, IP3, respectively), cAMP and progesterone in small luteal cells. LiCl (10 mM) enhanced inositol phosphate accumulation in response to LH but had no effect on LH-stimulated cAMP or progesterone accumulation. Time course studies revealed that LH-induced increases in IP3 and cAMP occurred simultaneously and preceded the increases in progesterone secretion. Similar dose-response relationships were observed for inositol phosphate and cAMP accumulation with maximal increases observed with 1 10 micrograms/ml of LH. Progesterone accumulation was maximal at 1-10 ng/ml of LH. LH (1 microgram/ml) and hCG (20 IU/ml) provoked similar increases in inositol phosphate, cAMP and progesterone accumulation in small luteal cells. 8-Bromo-cAMP (2.5 mM) and forskolin (1 microM) increased progesterone synthesis but did not increase inositol phosphate accumulation in 30 min incubations. PGF2a (1 microM) was more effective than LH (1 microgram/ml) at stimulating increases in inositol phosphate accumulation (4.4-fold vs 2.2-fold increase for PGF2a and LH, respectively). The combined effects of LH and PGF2a on accumulation of inositol phosphates were slightly greater than the effects of PGF2a alone. In 30 min incubations, PGF2a had no effect on cAMP accumulation and provoked small increases in progesterone secretion. Additionally, PGF2a treatment had no significant effect on LH-induced cAMP or progesterone accumulation in 30 min incubations of small luteal cells. These findings provide the first evidence that gonadotropins stimulate the cAMP and IP3-diacylglycerol transmembrane signalling systems in bovine small luteal cells. PGF2a stimulated phospholipase C activity in small cells but did not reduce LH-stimulated cAMP or progesterone accumulation. These results also demonstrate that induction of functional luteolysis in vitro requires more than the activation of the phospholipase C IP3/calcium and -diacylglycerol/protein kinase C transmembrane signalling system. PMID- 2544771 TI - Magnesium alterations and pharmacokinetic data in gallium-treated lung cancer patients. AB - The dose of gallium chloride required to inhibit tumor growth after oral and chronic administration depends on the stage of the cancer disease and of the type of metastases. A dose regimen of 800 mg/24 h of gallium chloride will provide serum gallium concentrations greater than or equal to 600 micrograms/l in lung cancer patients with a small and limited disease. A dose of 1,400 mg/24 h is well tolerated in metastatic patients but may not be high enough to reach the desired serum gallium concentrations especially in patients with bone metastases. Radiotherapy and/or a chemotherapy will permit one to increase the serum gallium concentrations and the tumor gallium uptake by reducing the volume of the tumor. After chronic, oral administration of gallium a decrease in RBC Mg is noted. To avoid the Mg deficiency, the treatment must not be interrupted and may perhaps be decreased with care and slowly without resulting in a decrease of the serum gallium concentrations provided the treatment has been prolonged over a sufficient time to enable one to induce intratumor biological modifications and a decrease in the number of the malignant cells. Acute pharmacokinetic data are related to the histologic type of the tumor and may not be used to predict the serum gallium concentrations after chronic administration. The serum gallium concentrations required to inhibit the tumor growth may be higher in small cell lung carcinomas than in nonsmall cell lung carcinomas. Frequent Mg and Ga blood determinations are necessary to manage effective gallium treatment. PMID- 2544772 TI - Role of the endothelium and arginine peptides on the vaso-motor response of porcine internal mammary artery. AB - The long-term patency of the internal mammary artery (IMA) graft is of considerable interest owing to its extensive use in myocardial revascularization. The aim of the present study was to elucidate the role of endothelium in modulating the responses of the porcine IMA to several vasoactive drugs. Isolated ring segments of porcine IMA contracted in a reproducible and dose dependent manner to phenylephrine, potassium chloride and the thromboxane mimic U46619, but the responses to serotonin, histamine and ATP were significantly less prominent. Both acetylcholine and bradykinin elicited endothelium-dependent relaxation which was not inhibited by indomethacin, but by methylene blue, an inhibitor of soluble guanylate cyclase. These two endothelium-dependent drugs and two endothelium independent relaxing drugs, nitroprusside and nitroglycerin relaxed the IMA in a dose dependent manner which was associated with an elevation of cyclic GMP. The endothelium dependent vasodilator peptides such as bradykinin contain L-arginine in their sequence. Benzoyl derivatives of L-arginine but not L-arginine relaxed the IMA in a dose dependent manner. These data confirm and extend exploratory studies performed with a simpler vascular model which indicate that the precursor of endothelium derived relaxing factor (EDRF) is an arginine moiety. PMID- 2544773 TI - Endocrine effects of psychological stress associated with neurobehavioral performance testing. AB - Twenty-four healthy subjects were submitted to a computer-based performance evaluation system. The set of tests required sustained attention, and the last test was expressly designed to cause a moderate, acute psychological stress. Compared to baseline levels, both serum ACTH and beta-endorphins were increased after psychological testing in all subjects. Serum prolactin showed a slight and statistically nonsignificant decrease compared to baseline values. These results question the belief that psychological stress stimulates prolactin secretion, whereas it suggests that serum ACTH and beta-endorphins are reliable indicators of acute psychological stress. PMID- 2544774 TI - Identification and characterization of atrial natriuretic factor receptors in the rat retina. AB - The characteristics of atrial natriuretic factor (ANF) receptors where studied in rat retinal particulate preparations. Specific 125I-ANF binding to retinal particulate preparations was greater than 90% of total binding and saturable at a density (Bmax) of 40 +/- 8 fmol/mg protein with an apparent dissociation constant (Kd) of 6.0 +/- 2.0 pM (n = 3). Apparent equilibrium conditions were established within 30 min. The Kd value of 125I-ANF binding calculated by kinetic analysis was 4.0 pM. The Bmax of 60 +/- 10 fmol/mg protein and the Kd of 5 +/- 2 pM, calculated by competition analysis, were in close agreement with the values obtained from Scatchard plots or kinetic analysis. The 125I-ANF binding to retinal particulate preparations was not inhibited by 1 microM concentration of somatostatin, vasopressin, vasoactive intestinal peptide, adrenocorticotropin, thyrotropin releasing hormone, or leu-enkephalin. The rank order of potency of the unlabelled atrial natriuretic peptides for competing with specific 125I-ANF (101-126) binding sites was rANF (92-126) greater than rANF (101-126) greater than rANF (99-126) greater than rANF (103-126) greater than Tyro-Atriopeptin I greater than hANF (105-126) greater than rANF (1-126). Similar results have been obtained in peripheral tissues and mammalian brain, indicating that central and peripheral ANF-binding sites have somewhat similar structural requirements. Affinity cross-linking of 125I-ANF to retinal particulate preparations resulted in the labelling of two sites of molecular weight 140 and 66 kDa, respectively. This demonstration of specific high-affinity ANF receptors suggests that the peptide may act as a neurotransmitter or neuromodulator in the retina. PMID- 2544775 TI - Short-term delay in blood processing does not alter basal or isoproterenol stimulated cyclic AMP accumulation in human lymphocytes. AB - Isoproterenol-stimulated cyclic AMP (cAMP) accumulation in lymphocytes is an index of beta-adrenergic receptor functioning. It is not known if a delay between blood collection and subsequent cell preparation influences cAMP accumulation. We compared cAMP levels in lymphocytes processed from whole blood immediately after collection and following a 3.5 hour delay. There were no significant changes in either basal or stimulated levels of cAMP. This information may alleviate time constraints in certain experimental designs where immediate isolation of lymphocytes from whole blood may not be practical. PMID- 2544776 TI - Effect of melatonin on steroid production by rat adrenals under in vitro superfusion conditions. AB - In vitro superfusion of adrenals from male and female Wistar rats resulted in a gradual decline of the corticosterone content in the samples collected for a period of 80 minutes. ACTH administration either in the beginning or in the end of the perfusion period led to a marked increase in the level of this steroid. Contrary to this, progesterone content in the collected perfusates was constant throughout the experiments and was not influenced by ACTH. The quantities of testosterone secreted under these conditions were below the sensitivity of the assay. Perfusion with melatonin (200 pg/ml) did not affect corticosterone secretion, but resulted in a significant increase of the progesterone content in the recovered fractions. This effect was immediate and was followed by a decline in the progesterone concentration, observed during melatonin perfusion. PMID- 2544777 TI - [Scintigraphy of the myocardium in ischemic heart disease]. AB - Altogether 106 patients with different types of acute CHD (large and small local MI, unstable angina) and stable angina were investigated. Combined assessment of perfusion disorder permits differentiation of necrotic and ischemic myocardial lesions. A degree and type of RP accumulation corresponds to a size of necrotic myocardial lesion determined by means of biochemical markers of necrosis. Parallelism of myoglobin concentration, isoenzyme activity in the blood serum and the results of scintigraphy was revealed. In focal RP accumulation, myoglobin concentration reached maximum values; a moderate increase and decrease up to normal values in the absence of accumulation were observed in diffuse accumulation. Diffuse RP accumulation in patients with stable and unstable types of angina was indicative of transient perfusion disorders resulting from myocardial ischemia. PMID- 2544778 TI - [Visualization of subcutaneous metastases of hypernephroid cancer using 99mTc dimercaptosuccinic acid]. PMID- 2544779 TI - [A comparative evaluation of radionuclide and ultrasonic studies of the thyroid]. AB - Unlike radionuclide scanning (RNS) ultrasound investigation (USI) permits thyroid volume estimation in hyperplasia and euthyroid goiter. USI in nodular goiter gives an opportunity to reveal the sizes of a tumor, its macrostructure, topographic position with regard to the neck organs and vessels and is a method of choice in autoimmune thyroiditis. A considerable advantage of USI is a possibility to characterize the thyroid collateral lobe in toxic adenoma and to perform investigations over time. A positive feature of RNS is a possibility for differential diagnosis of "hot" and "cold" nodes of the thyroid, metastases to other organs and atypical localization of the gland. PMID- 2544780 TI - [Polychemotherapy of large cell carcinoma of the lung: a case of complete remission]. AB - A case of a patient with large cell pulmonary carcinoma is presented. Being inoperable, the patient was treated at first with multiple drug chemotherapy and the result was the disappearance of the pulmonary lesions. The unusual favourable therapeutic response and the protocols of polychemotherapy used are discussed. PMID- 2544781 TI - Identification of a DNA fragment in the genome of Bordetella pertussis carrying repeated DNA sequences also present in other Bordetella species. AB - A DNA fragment (3.8 kbp) which hybridized to repeated sequences in the genome of Bordetella species has been cloned from Bordetella pertussis chromosomal DNA. Eleven subclones were constructed from this fragment. They exhibited distinct inter-species hybridization patterns in genomic blots of each of the Bordetella used in the study. One subclone revealed intraspecies variability among B. pertussis strains and another did not hybridize to B. parapertussis. The 3.8 kbp DNA fragment possesses a middle sequence surrounded by repeated sequences organized in an opposite symmetrical orientation. The external inverted repeats of it hybridized to a 680 bp DNA sequence which is located about 800 bp upstream of the pertussis toxin operon. The novel structural organization of the 3.8 kbp fragment suggests the possibility that this DNA segment is an IS-like element which may have an important function in the expression of virulence determinants in Bordetella bacteria. PMID- 2544782 TI - HIV and HTLV-I infections in the Americas: a regional perspective. AB - With over 143,000 cases of AIDS reported to the World Health Organization from 145 countries and with an estimated 5 to 10 million people worldwide infected with HIV, AIDS has become firmly established as a global pandemic. In the region of the Americas over 100,862 cases of AIDS have been reported with indigenous transmission documented in 45 to 46 countries. While North America has the highest annual number of AIDS cases per population, with 72 cases/million, the Caribbean subregion has a disproportionately high number of cases, with annual rates as high as 200 to 300 cases/million population for some countries. Despite differences in absolute number of cases, there has been a remarkable similarity in the temporal rate of increase of AIDS in the countries of the Americas, reflecting delayed introduction of the virus to some areas with an early exponential increase similar to that observed initially in the United States. Although the modes of transmission of HIV are the same throughout the region, evidence of increasing bisexual and heterosexual transmission, particularly in the Caribbean subregion, has resulted in a lower male-to-female ratio of AIDS cases and increased perinatal transmission. Clinically, a resurgence of diarrheal diseases, respiratory infections, and tuberculosis has been documented in association with HIV infection in many tropical countries of the Americas. With relatively high rates of HTLV-I infection already established in the Caribbean subregion, the overall public health problems of the Americas will be markedly potentiated by further spread of these 2 human retroviruses. If HIV infection continues to penetrate the poor and less advantaged populations in Latin America and the Caribbean, the potential exists for a massive epidemic in the Americas that may rapidly parallel the situation in Africa. PMID- 2544783 TI - Glyburide but not ciglitazone enhances insulin action in the liver independent of insulin receptor kinase activation. AB - To test the hypothesis that sulfonylureas enhance insulin action by activating the insulin receptor tyrosine kinase, the effects of glyburide, a second generation sulfonylurea, and ciglitazone, a nonsulfonylurea hypoglycemic agent, were determined in primary cultures of rat hepatocytes on insulin action and insulin receptor structure and function. Twenty hours of preincubation with glyburide (1 microgram/mL) resulted in increased insulin (1 X 10(-7) mol/L) stimulation of [14C] acetate incorporation into lipids and [14C] alpha aminoisobutyric acid uptake without any change in basal activity. Ciglitazone (1 microgram/mL) was without any effect. Glyburide's actions were mediated without altering the following: (1) 125I-insulin binding; (2) the electrophoretic mobility of the affinity labeled alpha-subunit or the autophosphorylated beta subunit of the insulin receptor; and (3) the insulin-stimulated insulin receptor kinase activity using histone or the beta-subunit of the insulin receptor as phosphoacceptors. These data suggest that the action of sulfonylureas is distal to the insulin receptor tyrosine kinase. Ciglitazone in vitro is ineffective in the liver, which suggests the peripheral tissues as the possible site of action. PMID- 2544784 TI - Increased glucose transport by human fibroblasts with a heritable defect in insulin binding. AB - Insulin and IGF-I binding and their regulation of hexose transport were evaluated in skin fibroblasts cultured from a family (Atl) whose proband had leprechaunism, hypoglycemia, and severe insulin resistance. High affinity insulin binding to proband Atl cells was absent, and partially, but equally, impaired in fibroblasts from his related parents. IGF-I binding to his cultured fibroblasts was within the normal range. Cells from proband Atl had insulin receptor mRNAs similar to control fibroblasts. 3-O-Methyl-D-glucose (OMG) transport by proband Atl was threefold higher than in control fibroblasts (37.7 v 7.6-11 nmol/mL/s) and was insulin-insensitive. Proband Atl fibroblasts had a threefold increase in the Vmax for OMG entry and a concomitant increase in the number of D-glucose-inhibitable cytochalasin B binding sites on their plasma membrane. Similar levels of glucose transporter mRNA were observed in control and proband Atl fibroblasts. These results suggest that fibroblasts from patient Atl have a genetically transmitted mutation in the alpha subunit of their insulin receptor. In the homozygous affected proband, this mutation impairs insulin binding and causes elevated, insulin-insensitive glucose transport. The dysfunction resulting from this mutation is similar to that introduced in Chinese hamster ovary cells by transfection with a truncated alpha subunit. PMID- 2544786 TI - Kinetics of herpes simplex virus photoinhibition by haematoporphyrin in both diploid and heteroploid cells. AB - The kinetics of inhibition of herpes simplex virus type 1 (HSV 1) on both diploid (CEF) and heteroploid cells (HEp2) by light-irradiated haematoporphyrin (HP) was studied. The inactivation of HSV1 by HP was drug-dose dependent and light irradiation dependent; the viruses grown in heteroploid cells being in all cases more sensitive to inhibition than viruses grown in diploid cells. Cell toxicity by HP was markedly more evident on HEp2 cells than on CEF. The highest viral sensitivity to photodynamic inactivation by HP was found to be between the 4th and the 5th hour after cell infection, when the viral DNA synthesis is at its peak and before it is incorporated into complete virions. Microfluorometric and spectrofluorometric assays revealed that virus infected cells always take up more HP than uninfected cells, and heteroploid cells incorporated more HP than diploid cells. The possibility that an increased uptake of HP and modifications of the cell micro-environment in virus infected cells could account for the viral inhibiting properties of HP, is discussed. PMID- 2544785 TI - Variation in herpes simplex virus type 1 glycoproteins produced in kidney cells from different species. AB - Viral glycoproteins from herpes simplex virus, type 1 (HSV-1) infected NBL-1, Vero, and BHK-21 cells were labelled with 14C-glucosamine and studied by SDS-PAGE and Con-A chromatography. SDS-PAGE analysis demonstrated differences in the number and molecular weight of glycoproteins from these cells. Con-A chromatography resulted in similar binding of glycoproteins from NBL-1 and Vero cells of 10.5 and 18.6%, respectively, whereas BHK-21 cells showed binding of 65%. These studies indicate that HSV-1 glycoprotein oligosaccharide processing is variable in kidney cells of different species. PMID- 2544787 TI - Augmentation of helper virus replication in the presence of defective retrovirus. AB - The interaction between defective spleen focus-forming virus (SFFV) and helper virus(es) in Friend virus (FV) complex has been assumed to be one-way, with the helper virus complementing SFFV by supplying necessary virion components. To test this assumption the expression of both SFFV and helper virus in partially congenic mice which differ at the Fv-2 locus, a gene that specifically controls susceptibility to SFFV, was analysed. When the mice were infected with LLV (a strain of Friend SFFV-free helper virus), there was no detectable effect of Fv-2 genotype on LLV expression as tested by virus infectivity in the XC plaque assay or by quantitative viral antigen analysis in an immunoprecipitation assay. However, after infection with FV complex there was an amplification of LLV (as well as SFFV) synthesis in Fv-2s as compared with Fv-2r hosts. To determine whether the increased LLV synthesis in Fv-2s mice was due to an increased population of susceptible target cells as a result of SFFV infection and/or transformation, the ratios of LLV-infected cells in the spleens of LLV- and FV infected Fv-2s hosts in an infectious centre assay, were compared. Since the percentage of LLV-infected cells was equivalent in both instances, the higher rate of LLV synthesis after infection with FV complex was presumably due to intrinsic properties of SFFV-infected erythroid cells. PMID- 2544788 TI - Advances in the understanding and management of the irritable bowel syndrome. PMID- 2544789 TI - [Platelet activating factor and platelet activating factor antagonists. New methods in asthma therapy?]. PMID- 2544790 TI - [Receptor diseases. 2: Hormone and lipoprotein receptors]. PMID- 2544791 TI - Compensation for beam intensity fluctuation in determination of P(ion) the ion recombination correction factor for ionization chambers, by the two-voltage technique. AB - We have developed a method of compensation for fluctuations in beam intensity that may occur during measurement of Pion, the ion-recombination correction factor, by the two-voltage technique. The method requires signals proportional to beam intensity during measurement. We used a parallel-plate ionization chamber, whose Pion was known, and a vacuum chamber to obtain signals that were proportional to the beam intensity. Experiments were conducted using pulsed proton beam providing doses that ranged from 0.16 to 0.01 cGy/pulse. The value of Pion of a thimble ionization chamber was measured. With these measurements, the validity of the method which we proposed for pulsed beam was verified experimentally. PMID- 2544792 TI - The use of variable grid spacing to accelerate dose calculations. AB - Planning radiation therapy using three-dimensional patient data is a very time consuming process with current hardware and software. When calculating a three dimensional dose distribution, the standard technique is to cover the volume of interest with a uniformly spaced matrix of points at which the dose is calculated. It is obvious that the dose is usually quite slowly varying in a large proportion of the region of interest; namely, in those regions which are either well inside or well outside the geometrical boundaries of the field. We have developed an algorithm which allows us to reduce the number of calculation points, and hence the time of calculation of the entire dose distribution, manyfold. We use a nonuniform grid of calculated points, based on the fact that the only regions which are troublesome for accurate dose interpolation are those in which large values of the second derivative of the dose as a function of position occur. We demonstrate that, at most grid points, the dose can be determined without decreasing accuracy below acceptable limits by simple linear interpolation between grid points much further apart than is usual in conventional techniques. We investigated our algorithm for one-, two-, and three dimensional examples and for Co-60, 25-MV photon, and 160-MV proton beams. In situations for which an accuracy of about 1% in dose and 1.6 mm in position was desired, we found gain factors for the number of points needing direct calculation of approximately 3 (one-dimension), 6 to 10 (two-dimensions) and 16 (three-dimensions). PMID- 2544793 TI - [Complete assignment of signals in 1D and 2D H-NMR spectra of a 17-member oligonucleotide, a model symmetrical analog of lambda operators]. AB - A synthetic analog of lambda phage operators, a symmetric duplex of oligonucleotides, was studied by 1H NMR at 400 MHz. Signals in the spectrum of imino protons of the duplex in H2O were assigned based on the results of NOE experiments and temperature dependences. Resonance assignment of the non exchangeable protons of bases and deoxyribose was performed by analysing the NOESY spectrum obtained in the single experiment. The results indicate that the major part of the duplex has a conformation similar to the B-form of DNA, and the region of the central non-complementary base pair exhibits deviations from the regular structure. PMID- 2544794 TI - [Topoisomerase I is connected with the regulatory zone of transcripted active mini-chromosomes of the SV40 virus]. AB - Proteins bound to SV40 DNA in sarkosyl-treated nuclei have been studied. The major component is topoisomerase I, a 60-70 kDa protein, which possesses a strong DNA-nicking activity in the presence of detergents. An SV40 fraction containing tightly bound proteins constitutes 2-3% of the total nuclear SV40 DNA and is enriched in transcriptionally active DNA as monitored by distribution of RNase resistant in vivo pulse labelled RNA. A small SV40 DNA fraction which is relaxed due to covalent binding of topoisomerase I upon sarkosyl treatment of isolated nuclei is also enriched in transcriptionally active DNA. In vivo topoisomerase I cleavage sites on SV40 DNA have been located by indirect end-labelling. Two preferential binding sites have been detected, both in the SV40 regulatory region. PMID- 2544795 TI - [Location of determinants of stability to neomycin and kanamycin as well as DNA segments, responsible for amplification in Streptomyces plasmids pSU3 and pSU10]. AB - The S. rimosus amplifying sequence AUD-Sr1 encodes kanamycin and neomycin resistance, defined in the case of neomycin by aminoglycoside phosphotransferase. Its cloning on plasmid SLP1.2 makes possible the co-amplification of the obtained hybrid plasmids in S. lividans. In our study the regions responsible for resistance to aminoglycoside antibiotics and the capacity for amplification the two hybrid plasmids pSU10 and pSU3 were determined. Experiments on subcloning of the AUD-Sr1 sequence fragments on vector pIJ702 revealed localization of kanamycin and neomycin resistance determinants between PvuII(6) and BglII(7) on the AUD-Sr1 sequence fragments of 2.0 kb length. Two regions responsible for amplification of the hybrid plasmids were detected with deletion and insertion mapping. The first region is localized in the region of the plasmid SLP1.2 BamHI site and the second region is localized on the PstI(4)-PvuII(6) of the AUD-Sr1 sequence fragment of 1.1 kb length. PMID- 2544796 TI - [Isolation of nucleotide sequences, possessing enhancer functions from the Saccharomyces cerevisiae genetic library]. AB - We propose a method for isolation of enhancer-like sequences from a yeast genomic library. The method was used to identify DNA inserts capable of increasing bacterial bla gene expression when located downstream from the transcription initiation site. Plasmids carrying different fragments of one such insert were used to localize the enhancer-like function on a 1.2 kb fragment. PMID- 2544797 TI - [Selective inhibition of 3'-5'-exonuclease activity of DNA-polymerase I from Escherichia coli by a fluoride ion]. AB - The effect of NaF on the enzymatic activities of the large fragment of E. coli DNA polymerase I (Klenow enzyme-KE) with different DNA-substrates was studied. It was shown that fluoride ion at concentrations of 5-10 mM efficiently inhibits the 3'----5' exonuclease activity of KE but does not affect the polymerase activity of the enzyme. Selective inhibition of the 3'----5' exonuclease activity of KE is Mg-dependent and is observed with double- or single-stranded DNAs. In reaction with the 14-mer oligonucleotide annealed with single-stranded phage M13 DNA the enzyme was found not only to perform the exonucleolytic hydrolysis of the primers but to catalyse also a limited elongation of some primers, adding a few nucleotide residues in the absence of exogenous dNTP. The primer elongation is inhibited by inorganic pyrophosphatase and is stimulated by micromolar concentrations of exogenous pyrophosphate thus suggesting a possible role of PPi contamination in dNTP generation via pyrophosphorolysis. Traces of precursors in DNA preparations obtained by generally employed methods may serve as another source of nucleotides for the primer elongation. PMID- 2544798 TI - [Organization of rRNA genes in various tribes of plants in the cereal family (Poaceae barnh.)]. AB - The organization of 18S and 26S rRNA in 14 plant species, belonging to 8 tribes of the cereal family was studied. In rDNA of all the cereals studied, except maize and reed, the similar character of localization of nucleotide sequences, recognized by restrictases BamHI and EcoRI in 18S and 26S rRNA genes was revealed. The structural organization of rDNA of sainfoin (Papilionaceae) was shown to differ from genes, coding for high molecular rRNA in cereals. The primary structure of subrepeat of non-transcribed rDNA spacer of diploid wheat Tr. urartu, consisting of 132 base pairs was determined. The given subrepeat was hybridized with BamHI-fragments of DNA from cereals and sainfoin. It is shown to hybridize with rDNA of all the cereals studied, and it hardly hybridizes with rDNA of maize and sorghum, but doesn't hybridize with rDNA of sainfoin. The conclusion is made that the size polymorphism of restriction fragments in the coding rDNA region and the level of similarity of subrepeats of rDNA of the non transcribed spacer may help to reveal the phylogenetic affinity of plants, belonging to different tribes within one family. PMID- 2544799 TI - [Features of the chromatin structure of erythrocytes depending on the properties of lysine-rich histones]. AB - A comparative analysis of chromatin from erythrocytes of frog, trout and hen has been performed in correlation with properties of the nucleosomal linker histones of H1 family. In the nucleosomes from frog erythrocytes the linker histone is represented by H1(0)-like variant with amino acid sequence highly homologous to that of the hen histone H5, however the arginine content in the proteins differs (3 mol% in the frog erythrocyte H1 and 12 mol% in the hen erythrocyte H5). On the other hand histone H5 from trout being significantly different in the primary structure from the hen histone H5 is at the same time rich in arginine (9 mol%). The nucleosomal repeat length, estimated by using agarose gel electrophoresis is 201, 213 and 213 b.p. in erythrocyte chromatin from frog, trout and hen, correspondingly. Chromatin packing density in fixed nuclei from erythrocytes of frog, trout and hen as determined using cytophotometric measurements is 0.144, 0.444 and 530 pg/mu 3, correspondingly. The data support the previously made suggestion that the increase in arginine content in nucleosomal linker proteins is connected with the increase of chromatin compaction in the nuclei and elongation of the linker in the nucleosome. PMID- 2544800 TI - Multiple cis-acting DNA regulatory elements mediate hepatic angiotensinogen gene expression. AB - Angiotensinogen is the glycoprotein precursor of angiotensin II, an octapeptide hormone important for the regulation of blood pressure and volume homeostasis. The gene encoding angiotensinogen is expressed in liver and several other tissues, providing a model gene for understanding the role of cis-acting DNA control elements and trans-acting factors in tissue-type specific gene expression. To identify DNA control elements in the rat angiotensinogen gene we prepared an array of fusion genes consisting of either 5' or 3'-deleted sequences of the 5'-flanking region of the gene linked to a firefly luciferase reporter gene and analyzed the relative cellular specificity of expression of these genes after their introduction into hepato-carcinoma cells (Hep G2) that do express and placental cells (JEG-3) that do not express the endogenous angiotensinogen gene. Six transcriptionally active elements were found within 688 base pairs of 5' flanking DNA. The interactions of DNA binding proteins with these elements was demonstrated by their specific protection to digestion with DNase I in the presence of liver cell extracts. The orientation and spatial requirements for transcription of two of the elements were analyzed further by the construction and expression of synthetic oligonucleotide cassettes incorporating the sequences of these elements when linked to a homologous (angiotensinogen) or a heterologous Simian virus 40 promoter and enhancer. One element located between 60 and 108 base pairs from the start of gene transcription functioned either as a silencer or an enhancer of transcription (SOAP box element), depending upon the distance from the angiotensinogen and viral gene promoters. Moreover, the SOAP box element demonstrated enhancer activity in JEG-3 cells when linked to the Simian virus 40 early promoter. An oligonucleotide mutation of the SOAP box element interfered with protein binding in a gel mobility shift assay and this mutant was transcriptionally inactive in both homologous and heterologous promoters. These observations indicate that expression of the angiotensinogen gene in liver cells is coordinately regulated by multiple cis-acting elements that interact with DNA binding proteins. PMID- 2544801 TI - Interferon-gamma inhibits steroidogenesis and accumulation of mRNA of the steroidogenic enzymes P450scc and P450c17 in cultured porcine Leydig cells. AB - The inhibitory effects of recombinant porcine interferon-gamma (IFN gamma) on human CG (hCG)-stimulated testosterone production, and on mRNA concentrations of cholesterol side-chain cleavage (P450scc) and 17 alpha-hydroxylase/C17-20lyase (P450c 17) were investigated using porcine primary Leydig cell culture as a model. After preincubation of Leydig cells for 24 h with 1000 pM IFN gamma, hCG stimulated (10 ng/ml, 2 h) testosterone production was inhibited by 50%, whereas no significant changes were seen in hCG-stimulated cAMP production. Incubation with 10 microM 5-cholestene-3 beta,22(R)-diol or 10 microM 5-cholestene-3 beta,20 alpha-diol together with hCG (10 ng/ml, 2 h) reversed most of the inhibitory effect of IFN gamma, suggesting that IFN gamma inhibits P450scc activity, possibly by inhibiting the substrate (cholesterol) availability for P450scc. Incubation with IFN gamma also decreased basal concentrations of P450scc (45%) and P450c 17 (35%) mRNA, although these changes probably did not contribute to the decreased testosterone production. Long-term treatment with hCG (100 ng/ml, 24 h) increased P450scc mRNA (3- to 4-fold) and P450c 17 mRNA (4- to 5-fold) concentrations. Simultaneous treatment with IFN gamma attenuated these hCG induced increases in P450scc mRNA (50%) and P450c 17 mRNA (40-100%) concentrations, as well as in testosterone production (77%). This inhibition of testosterone production could only be partly reversed by the hydroxylated cholesterol derivatives. This suggests that in addition to possible suppression of cholesterol availability, decreased P450scc and/or P450c 17 activities (through decreased mRNA concentrations) were also involved in the IFN gamma suppressed steroidogenic capacity of porcine Leydig cells during long-term hCG stimulation. PMID- 2544802 TI - A role for the insulin-like growth factor II/mannose-6-phosphate receptor in the insulin-induced inhibition of protein catabolism. AB - Insulin and insulin-like growth factor (IGF)-I inhibit intracellular protein degradation in a variety of different cell types. In the present studies, the IGF I-induced inhibition of protein metabolism in Chinese hamster ovary (CHO) cells was found to be blocked by polyclonal antibodies to the IGF-II/mannose-6 phosphate phosphate (Man-6-P) receptor, but not by control immunoglobulin. In contrast, these antibodies had no effect on the ability of IGF-I to stimulate glucose uptake in the same cells. The antibodies to the IGF-II/Man-6-P receptor also inhibited the effect of IGF-I and insulin on protein catabolism in human foreskin fibroblasts and human hepatoma cells, respectively. Moreover, CHO cells overexpressing a cDNA coding for the IGF-II/Man-6-P receptor were found to exhibit an increased effect of insulin on protein catabolism. In contrast, the insulin stimulation of glucose uptake is the same in these transfected cells as in the parental CHO cells. These results implicate the IGF-II/Man-6-P receptor in the insulin- and IGF-I-induced inhibition of protein catabolism. PMID- 2544803 TI - [Isolation of DNA fragments of the barley genome, supporting autonomous regulation of plasmids in Saccharomyces cerevisiae]. AB - BamHI fragments of the barley genomic DNA were cloned in Escherichia coli cells on the vector plasmid YIp5 carrying the URA3 gene of Saccharomyces cerevisiae. Yeast cells were transformed by individual plasmid DNA preparations from each clone selected. Approximately 10% of the studied plasmids are able to replicate in yeast cells. Five barley DNA fragments which could support replication of recombinant plasmids in yeast cells belong to moderately repeated DNA and are dispersed through the chromosomes. PMID- 2544804 TI - Mammalian cell genotoxicity: a major role for non-DNA targets? PMID- 2544805 TI - AIDS: US forecast too low? PMID- 2544806 TI - Diversity of gamma delta T-cell receptors on murine intestinal intra-epithelial lymphocytes. AB - The search for the genes encoding the T-cell receptor (TCR) alpha- and beta subunits revealed a third gene gamma which shares with the alpha- and beta-genes several properties including somatic rearrangement. This gene, together with a fourth rearranging gene delta, encodes a second type of T-cell receptor, TCR gamma delta. Although TCR gamma delta-bearing T cells constitute a relatively minor subpopulation in the thymus and in peripheral lymphoid organs, they are the major lymphocytes of epidermis (dendritic epidermal cells or DEC) and of intestinal epithelium (intestinal intraepithelial lymphocytes or IEL) in mice, suggesting that at least some gamma delta T cells are important in the surveillance of a variety of epithelia. It was recently reported, however, that the TCR gamma delta on DEC has essentially no structural diversity, implying that the putative ligand is monomorphic. As this finding, if generally applicable, poses severe restrictions on the origin of the ligand, we investigated the diversity of the TCR on the second major epithelium-associated gamma delta T cells, namely IEL from mice. We report here that by contrast with the DEC gamma delta, the IEL gamma delta TCR are structurally diverse. PMID- 2544807 TI - Cloning of murine alpha and beta retinoic acid receptors and a novel receptor gamma predominantly expressed in skin. AB - In addition to having profound effects on embryonic pattern formation, retinoic acid (RA) has striking effects on differentiation and maintenance of epithelial cells in vivo and in vitro Skin is a major target organ for retinoids both in its normal and pathological states. The discovery of two human nuclear receptors for RA (hRAR alpha and hRAR beta) acting as transcriptional RA-inducible enhancer factors has provided a basis for understanding how RA controls gene expression. To investigate the specific role that RARs might play during development and in adult tissues, we have cloned the mouse RAR alpha and RAR beta (mRAR alpha and mRAR beta). Their amino-acid sequences are much more homologous to those of hRAR alpha and hRAR beta, respectively, than to each other, which suggests strongly that RAR alpha- and beta-subtypes have different functions. Most interestingly we have discovered a novel RAR subtype (mRAR gamma) whose expression in adult mouse seems to be highly restricted to skin, whereas RAR alpha and RAR beta are expressed in a variety of adult tissues. Furthermore, both mRAR alpha and mRAR gamma RNAs are readily detected in undifferentiated F9 embryocarcinoma (EC) cells, whereas mRAR beta messenger RNA is induced at least 30-fold in RA differentiated F9 cells. PMID- 2544808 TI - Control of transmembrane lipid asymmetry in chromaffin granules by an ATP dependent protein. AB - The Ca2+-dependent binding of annexin proteins to secretory granule membranes seems to be involved in the early stage of exocytosis. Binding studies have shown that these proteins have a specificity for phosphatidylserine (PtdS) interfaces. Furthermore, aminolipids are necessary for contact and fusion between lipid vesicles or between liposomes and chromaffin granules. Thus, PtdS must be present on the granule outer (cytoplasmic) monolayer. We report here that chromaffin granules possess a mechanism to maintain PtdS orientation, comparable to the ATP dependent aminophospholipid translocase from human erythrocytes. The translocase, in granules, selectively transports PtdS from the luminal to the cytoplasmic monolayer, provided the incubation medium contains ATP. As this protein shares several properties with the granule vanadate-sensitive ATPase II, we infer that this ATPase, of relative molecular mass 115,000, is the protein responsible for aminophospholipid translocation. This is the first evidence for an ATP-dependent specific phospholipid 'flippase' in intracellular organelles. PMID- 2544809 TI - Increased viral pathogenicity after insertion of a 28S ribosomal RNA sequence into the haemagglutinin gene of an influenza virus. AB - The haemagglutinin glycoprotein HA of influenza viruses is responsible for the attachment of the virus to neuraminic acid-containing receptors at the cell surface and subsequent penetration by triggering fusion of the viral envelope with cellular membranes. To express full activity of the newly synthesized precursor, HA has to be modified by post-translational proteolytic cleavage into the polypeptides HA1 and HA2 by cellular enzymes. If proteases suitable for cleavage are not present in the host cell, the resulting virus particles are non infectious. During adaptation of the apathogenic influenza virus A/turkey/Oregon/71 to chicken embryo cells, which are not permissive for HA cleavage, we obtained an infectious virus variant with increased pathogenicity. Sequence analysis revealed that during adaptation 54 nucleotides were inserted into the HA gene; their sequence corresponds to a region of the 28S ribosomal RNA. This insertion is probably responsible for increased cleavability of HA, as well as for infectivity and pathogenicity of the adapted virus. PMID- 2544810 TI - Amiodarone is a potent calmodulin antagonist. AB - The possible interaction between amiodarone, a potent antiarrhythmic and antianginal agent, and calmodulin (CaM) was investigated by three avenues of approach: (a) Effect of amiodarone on cardiac and vascular Ca2+/calmodulin activated cyclic nucleotide phosphodiesterase (CaM-PDE); (b) Effect on the CaM activated (Ca2+ + Mg2+)-ATPase from human erythrocytes; (c) Direct interaction between amiodarone and calmodulin measured by the effect of the drug on the fluorescence of 9-anthroylcholine (9AC) bound to calmodulin. Results show that amiodarone did not interact with basal activities of CaM-PDE and other isolated CaM-insensitive PDE forms as well as with (Ca2+ + Mg2+)-ATPase. Amiodarone inhibited calmodulin-activation of aortic CaM-PDE (Ki = 650 nM, substrate cGMP) and calmodulin-activation of erythrocyte ghosts (Ca2+ + Mg2+)-ATPase (IC50 = 4.5 microM) in an apparently competitive manner. Amiodarone decreased the fluorescence of the hydrophobic probe 9AC bound to calmodulin (IC50 = 5 microM). It is concluded that amiodarone is a potent calmodulin antagonist. PMID- 2544811 TI - Evidence that clomethiazole interacts with the macromolecular GABA A-receptor complex in the central nervous system and in the anterior pituitary gland. AB - Clomethiazole (CLOM) is known to be an anticonvulsant drug and has been also reported to decrease serum prolactin (PRL) in humans. Both effects may be mediated by an enhancement of gabaergic transmission. In order to determine if (CLOM) interacts with GABA metabolism and/or at the GABA receptor level, we studied its effect on PRL release and on the binding of various compounds that interact with the GABAA-benzodiazepine-receptor complex. Intraperitoneal (IP) administration of CLOM to rats significantly decreased PRL levels, and this effect was antagonized by IP administration of bicuculline, an antagonist of the GABAA receptor. In vitro, the inhibitory effect of muscimol on PRL release from rat hemiadenohypophysis was potentiated in a dose-dependent manner by preincubation with CLOM. This effect was antagonized by picrotoxin (10(-6) M). On the other hand, CLOM had no effect on GABA metabolism and did not compete with GABAA, GABAB or benzodiazepine binding sites in cortical membranes. CLOM competed, however, with the picrotoxin binding site labelled with [35S] butylbicyclophosphorothionate (TBPS), at an IC50 value of 1.2 x 10(-4) M, which is in the same range as some barbiturates. These results concerning PRL release and binding experiments with cortical membranes suggest that CLOM interacts with the picrotoxin/barbiturate site of the GABAA-receptor-chloride channel complex. PMID- 2544813 TI - [Papillomavirus and carcinoma of the uterine cervix]. PMID- 2544812 TI - Sodium channel-blocking properties of flecainide, a class IC antiarrhythmic drug, in guinea-pig papillary muscles. An open channel blocker or an inactivated channel blocker. AB - Effects of flecainide (a class IC antiarrhythmic drug) on the maximum rate of rise (Vmax) of action potentials (APs) were studied in guinea-pig papillary muscles, with special reference to their time, voltage, and action potential duration (APD) dependence in the presence and absence of nicorandil. Nicorandil was used to shorten APD, i.e., the time period of inactivation state of sodium channels. APs were recorded from the preparations using standard microelectrode techniques. Flecainide (5 mumol/l) reduced Vmax without changing resting potential, AP amplitude, APD50, and APD90 examined at 1 Hz. The drug shifted the normalized Vmax-membrane potential curve (examined at 1/60 Hz) in the hyperpolarizing direction by 3.1 +/- 0.8 mV (n = 6) (voltage dependence). The drug caused a frequency-dependent reduction of Vmax at greater than or equal to 0.1 Hz, developed a use-dependent reduction of Vmax at 1 Hz with an onset time constant of 11.7 +/- 0.4 s (n = 6), and slowed the recovery process of Vmax, whose resultant recovery time constant was 19.9 +/- 1.2 s (n = 6) (time dependence). These flecainide-induced time-dependent reductions of Vmax were not antagonized by nicorandil (1 mmol/l) which shortened APD to about 1/4 of control (APD independence). These results suggest that flecainide is primarily an open channel blocker because its channel-blocking actions are independent of APD or the time period of inactivation. PMID- 2544814 TI - [The treatment of acute gastroenteritis in children in The Netherlands]. AB - A national survey on the treatment of acute gastroenteritis in children was conducted among 10% of the Dutch general practitioners and 10% of the paediatricians. Oral Rehydration Solution was used by less than 40% of the general practitioners, while 95% of the paediatricians used it (nearly) always. About a quarter of the general practitioners used antidiarrhoeal medication in contrast to the paediatricians who hardly or never used these drugs. The refeeding period after the acute rehydration therapy showed great difference. The need for a uniform dietary advice was felt by nearly half the general practitioners. It is concluded that the internationally accepted primary treatment with O.R.S. has insufficient impact on the Dutch general practitioners. Greater knowledge on the unique features of this method is needed. The dietary advice after the primary treatment needs more uniformity. PMID- 2544815 TI - Intraarterial infusion of carboplatin in the treatment of malignant gliomas: a phase II study. AB - Twenty-three previously treated patients with malignant gliomas were included in this phase II study of carboplatin (400 mg/m2) given as an intraarterial infusion every 4 weeks. Five patients (26% of 19 evaluable) achieved a partial response for 3 to 10 months and 5 patients presented a stabilization for 2 to 7 months. Toxicity was mild in most patients, with nausea, vomiting and myelosuppression being the most frequent side-effects. No renal or auditory toxicity was observed. One patient developed central nervous toxicity and another one a reversible fall in visual acuity. Finally, carboplatin infused by the intraarterial route seems to be an active drug, less toxic than cisplatin, in malignant gliomas, and further trials are warranted. PMID- 2544816 TI - [Neurologic complications caused by anticoagulants and fibrinolytic agents]. AB - We examined 126 patients, of whom 54 had suffered brain hemorrhage, 33 subdural hematoma, 18 subarachnoid bleeding, nine spinal hemorrhage and twelve hematoma with peripheral deficits. Neurologic disorders were seen in patients subject to therapy with coumarins 2-4 years after initiating therapy, whereas hemorrhages under heparine and streptokinase regimes were observed after only a few days. Clotting values were below a therapeutic range in only some of the cases. A clinical diagnosis should be verified by CT scanning. Blood clotting normally recovers after discontinuation of anticoagulation or fibrinolysis. Surgery will improve the prognosis in many cases of subdural hematoma or spinal hemorrhage; conservative treatment in cases of intracerebral, subarachnoid, and peripheral hemorrhage. A 70-percent lethality was recorded for patients suffering a cerebral hemorrhage. More than a third of patients with subarachnoid bleeding and less than a third of cases with subdural hematoma died. While peripheral lesions tended to improve, spinal hemorrhage often resulted in irreversible paraplegia. By respecting contra-indications for anticoagulation therapy and limiting the duration of such regimes the risk of hemorrhage within the CNS and other nerve structures may be reduced. Interactions with other drugs precipitating clotting disorders should also be taken into account. PMID- 2544817 TI - Enalapril attenuates glomerular hyperfiltration following a meat meal. AB - It has been shown that the glomerular filtration rate increases after a meat meal. We examined in humans whether enalapril, which has been shown to decrease glomerular capillary pressure in rats with chronic renal failure, could attenuate the renal response to a meat meal. Twelve healthy volunteers were studied after an oral protein load, 1.5 g/kg body weight, as lean cooked beef meat, and on a separate day, after eating the same meal with prior oral intake of enalapril. On the control day, creatinine clearance increased from 114.3 +/- 4.7 before the meal to 137.1 +/- 4.7 ml/min/1.73 m2 after the meal (p less than 0.001). On the enalapril intake day, creatinine clearance increased from 113.7 +/- 5.6 before the meal to 128.3 +/- 5.8 ml/min/1.73 m2 after the meal (p less than 0.01). However, the mean increase in creatinine clearance was lower on the enalapril intake than on the control day (14.0 +/- 4.3 vs. 21.0 +/- 4.1%, p less than 0.05). Mean arterial pressure before the meal was lower on the enalapril intake day than on the control day (76.2 +/- 3.5 vs. 84.2 +/- 3.6, p less than 0.01). Likewise, postprandial mean arterial pressure was lower on the enalapril day compared with the control day (69.9 +/- 2.8 vs. 78.5 +/- 3.7, p less than 0.01). We conclude that enalapril blunts the hyperfiltration which follows a meat meal. PMID- 2544818 TI - Parameters of evaluation and correlation of renal phosphate handling and of parathyroid function in children. AB - The aim of the study was to establish normal values for parameters of parathyroid (PT) function in childhood and to examine whether a correlation could be found between these parameters and tubular phosphate (P) reabsorption. Values for PTH (carboxyl terminal) and urinary cAMP (UcAMP) throughout childhood were found to be significantly lower than in adults, with mean values of 0.3 ng/ml (30 pmol/l) and 1.96 nmol/dl glomerular filtrate (GF) respectively. A significant correlation was found between UcAMP and tubular P reabsorption (TP), expressed by the equation: TP = 5.5-(0.5 x UcAMP) mg/dl GF (= 1.78-(0.16 x UcAMP) mmol/dl GF, with a scatter of 10.6 mg/dl around the mean (+/- 1 SD). These data appear to permit the evaluation of primary tubular components in patients with defects of renal P handling, as opposed to those related to PT activity. PMID- 2544819 TI - Epstein-Barr-virus-associated lymphoproliferation arising in a renal allograft. PMID- 2544820 TI - Effects of a protein load in patients with early chronic renal failure before and after angiotensin II blockade. AB - We studied the effects of mid-term enalapril administration on protein-load induced renal responses in 10 patients with early chronic renal failure (serum creatinine 2.70 +/- 1.0 mg/dl). The oral protein load was performed twice, before and after a 10-day therapy with enalapril. Glomerular filtration rate (125I iothalamate clearance) rose from 22.5 +/- 10.6 to 60.1 +/- 32.8 ml/min after the protein load before enalapril; it did not change after the protein load during enalapril therapy. Percent fractional excretion of sodium, urinary osmolality and free water clearance were significantly affected only by the protein load before enalapril. Enalapril blunts the protein-load-induced changes in glomerular filtration rate and in tubular function; these effects might be mediated by angiotensin II blockade. PMID- 2544821 TI - Effects of physiological infusion of atrial natriuretic factor on healthy subjects and patients with the nephrotic syndrome. AB - We followed the renal and hormonal effects of physiological intravenous infusions of atrial natriuretic factor (ANF) in 6 water-loaded patients with nephrotic syndrome and 7 healthy subjects. Two of the patients had impaired renal function, 3 had active sodium retention, and none took drugs. The ensuing natriuresis, increase in plasma and urinary cyclic guanosine monophosphate and suppression of the renin-aldosterone axis were similar in normals and nephrotics. In both groups, significant increases in filtration fraction (inulin/PAH clearance) were observed, and in the nephrotics, major increases also occurred in both the absolute and fractional urinary albumin excretion. The renal and hormonal responses to ANF are not impaired in the nephrotic syndrome. PMID- 2544822 TI - A study of infantile motor neuron disease with neurofilament and ubiquitin immunocytochemistry. AB - We report a patient with infantile motor neuron disease who had pathologic findings consistent with multisystem degeneration. Although the muscle showed denervation atrophy and spinal anterior horn cells showed either atrophy or ballooning degeneration consistent with lower motor neuron disease, the infant was hypertonic and spastic. Degenerative changes were also detected in the dorsal root ganglia, cerebellum, and thalamus. Immunohistochemical studies showed a paucity of neurofilament (NF) staining in the corticospinal tract and accumulation of phosphorylated NF in ballooned neurons. Antibodies to ubiquitin immunostained ballooned neurons in the dorsal root ganglia, anterior horns, and thalamus. Accumulation of ubiquitinated and phosphorylated NF epitopes in degenerating neurons suggest that basic abnormalities in the neuronal cytoskeleton may be instrumental in the pathogenesis of this disorder. PMID- 2544824 TI - Neurotensin receptors in the human spinal cord: a quantitative autoradiographic study. AB - The anatomical localization of neurotensin receptors in the human spinal cord was examined in 12 cases aged 4-68 years using quantitative autoradiographic methods following the incubation of fresh, unfixed cryostat sections with 4 nM [3H]neurotensin. Characterization of the pharmacological specificity of the [3H]neurotensin binding sites in the human spinal cord from displacement studies with neurotensin and various neurotensin fragments indicated that, whereas 1.0 microM neurotensin and the carboxy-terminal fragment neurotensin almost completely displaced [3H]neurotensin binding (4 nM), the amino-terminal fragments neurotensin and neurotensin1-11 were weak inhibitors. This requirement for the carboxy-terminal fragment neurotensin is consistent with [3H]neurotensin binding to specific neurotensin receptors in the human spinal cord. In all cases the autoradiograms demonstrated that neurotensin receptors were distributed in a similar fashion in the gray matter of the cervical, thoracic, lumbar, sacral and coccygeal regions of the human spinal cord. At all 21 spinal levels examined, the highest density of neurotensin receptors was localized in lamina II of the dorsal horn. Within lamina II the receptors were especially concentrated in the deeper inner segment (IIi) where they formed a dense band lying immediately dorsal to lamina III. The density of receptors in this inner region of lamina II (23.5 fmol/mg) was almost double that in the outer segment of lamina II (12.2 fmol/mg), which showed the next highest density of receptors, and more than three times that in the adjacent lamina I (6.9 fmol/mg) and lamina III (7.1 fmol/mg). A moderate density of receptors was present in the intermediomedial (8.0 fmol/mg) and intermediolateral (8.0 fmol/mg) nuclei of lamina VII, and in lamina IX (4.4 fmol/mg). The density of labelling in the remaining laminae of the spinal cord was very low. These results indicate that neurotensin receptors are mainly localized in somatic and visceral sensory and motor regions of the human spinal cord and suggest that neurotensin may play a role in modulating sensory-motor functions in the human spinal cord. PMID- 2544823 TI - Haloperidol and cognitive shifting. AB - In this study haloperidol appeared to affect the performance on a selected category of cognitive tasks considered to represent shifting aptitude. A pretest- post-test design was used with two groups of subjects: 17 patients suffering from idiopathic spasmodic torticollis, and 17 controls who were matched for age and intelligence. The results are discussed in relation to previous findings on haloperidol and cognition, shifting disorder in Parkinson's disease and changes in behavioural organization found in animals with an experimentally induced dopaminergic hypoactivity. PMID- 2544825 TI - Ontogeny of somatostatin receptors in the rat brain: biochemical and autoradiographic study. AB - The ontogeny of somatostatin receptors in the rat brain has been studied by both membrane binding assays and in vitro receptor autoradiographic techniques. High levels of somatostatin binding sites were detected in brain of 15-day-old fetuses (E15). The pharmacological characterization of somatostatin binding sites and the regulatory effect of GTP on somatostatin binding at E15 suggest that somatostatin recognition sites correspond to authentic receptors. The values of maximal binding showed important variations throughout pre- and postnatal development. Globally, a marked increase in the total binding capacity was observed between E15 and postnatal day 8 (P8), with a transient fall at birth and P1. After P8, the concentration of somatostatin receptors progressively decreased and the weaning imposed at P21 accentuated the decline of receptor concentration. Although the density of somatostatin binding sites varied considerably, KD values did not change during brain development. Autoradiographic studies showed marked differences in the distribution of somatostatin receptors during ontogenesis. In the cortex, the cortical plate and the subplate zone appeared to contain high densities of binding sites from E15 to P1. However, the cortical layer which exhibited the higher labelling was the intermediate zone, located just beneath the subplate zone. On the contrary, the germinal epithelium bordering the lateral ventricle appeared virtually devoid of somatostatin binding sites. This laminar distribution of binding sites in the cortex disappeared from P4 to P8, in coincidence with the evolution of the underlying histological organization. At these stages, a homogeneous distribution was observed in almost all cortical layers, contrasting with the distribution of somatostatin receptors in the adult, which was restricted to layers IV-VI. In the cerebellar cortex, autoradiographic labelling was first seen at E15. After birth, the density of somatostatin receptors increased dramatically between P4 and P13, while, at P23, the labelling vanished in most lobes of the cerebellum. Taken together, these results show the early appearance of somatostatin receptors in the rat brain. The high density of somatostatin receptors observed in proliferative or pre-migratory areas suggests that somatostatin may be an important factor involved in the organization of the central nervous system. PMID- 2544826 TI - Autoradiographic localization of particulate cyclic AMP-dependent protein kinase in mammalian brain using [3H]cyclic AMP: implications for organization of second messenger systems. AB - Cyclic AMP's regulatory role as an intracellular second messenger is well established. In brain and other tissues, specific proteins that bind cyclic AMP have been shown to be the regulatory subunits of cystolic and particulate cyclic AMP-dependent protein kinases. This study of the autoradiographic localization of specific [3H]cyclic AMP binding revealed the heterogeneous distribution of particulate cyclic AMP-dependent protein kinase in the mammalian central nervous system. Specific [3H]cyclic AMP binding to tissue sections was of high affinity (KD = 60 nM) and saturable (Bmax = 5 pmol/mg protein). Purine and pyrimidine nucleotide analogues demonstrated inhibition constants against [3H]cyclic AMP binding consistent with the specific labelling of cyclic AMP-dependent protein kinase (e.g. 8'-bromo-cyclic AMP: IC50 = 130 nM; inosine 3',5'-cyclic monophosphate: IC50 = 1 microM; uridine 3',5'-cyclic monophosphate: IC50 = 60 microM). Variations in the levels of [3H]cyclic AMP binding presumably reflect the presence of differing amounts of particulate cyclic AMP-dependent protein kinase in different neuronal populations. Highest densities were associated with neuronal cell layers such as the pyramidal cells of the piriform cortex and hippocampus, and granule cells of the dentate gyrus and cerebellum. High levels of binding were also found in other cortical and limbic structures, while moderate levels were found in hypothalamic, thalamic and midbrain areas. Excitotoxic lesions confirmed the localization of the enzyme in hippocampal pyramidal cells and cerebellar granule cells. Localizations reported in this study are largely consistent with results obtained using immunohistochemical methods to label cyclic AMP-dependent protein kinases. Recently, [3H]forskolin, a potent and selective activator of adenylate cyclase, the enzyme responsible for the formation of cyclic AMP from adenosine 5'-triphosphate, has been used to localize the activated catalytic component of this enzyme in rat brain. Regions described as being intensely labelled with [3H]forskolin (e.g. basal ganglia, hilus of the dentate gyrus and molecular layer of the cerebellum) were found to be associated with relatively low [3H]cyclic AMP binding levels. These findings suggest a marked difference between the localization of the two related enzyme entities. However, the distribution of the enzymes is indirectly correlated as high levels of particulate cyclic AMP-dependent protein kinase are present in the soma of neurons with high concentrations of adenylate cyclase in their terminals. Alternatively, it is possible that [3H]forskolin localizes only a subpopulation of adenylate cyclase.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2544827 TI - Nuclear inclusions in innervated cultured muscle fibers from patients with oculopharyngeal muscular dystrophy. AB - We established monolayer muscle fiber cultures from muscle biopsies of 3 patients with oculopharyngeal muscular dystrophy (OPMD) who had characteristic intranuclear inclusions (INI-A) in their muscle fibers. Aneural cultures had normal morphology, except for a few muscle fibers that contained small vacuoles. Innervated cultures had large cytoplasmic vacuoles in a number of muscle fibers. Those muscle fibers were breaking easily, and could not be maintained longer than 2 months. Electron microscopy showed unusual intranuclear inclusions (INI-B) not previously reported in aneurally cultured muscle fibers of OPMD or in any normal or disease-control aneural or innervated cultured human muscle fibers. They resembled, but were not identical to, the INI-A, and they occurred in both the cultured fibers and the original muscle biopsies of all 3 patients. Our study demonstrate that (1) nuclear inclusions in OPMD reflect an intrinsic genetic defect; and (2) neuronal influence, advanced maturation, or both, seem to be essential for their induction in muscle fibers. PMID- 2544828 TI - Nerve growth factor receptor immunoreactivity in the neuronal perikarya of human sensory and sympathetic nerve ganglia. AB - We examined immunohistochemically the dorsal root ganglia, sympathetic ganglia, spinal cord, ventral and dorsal roots, and sciatic nerves obtained at autopsy from adult humans, using a monoclonal antibody against the human nerve growth factor receptor. We observed labelling in a granular pattern in the neuronal perikarya of dorsal root and sympathetic nerve ganglia. Ventral horn cells and axons were not labelled. PMID- 2544829 TI - Relative efficacy of intravenous methylprednisolone and ACTH in the treatment of acute relapse in MS. AB - To compare the efficacy of high-dose intravenous methylprednisolone with intramuscular ACTH in the treatment of acute relapse in multiple sclerosis, we undertook a double-blind, randomized, controlled study involving 61 patients. There was a marked improvement in both groups in the course of the study, but no difference between them in either the rate of recovery or the final outcome. High dose IV methylprednisolone is a safe alternative to ACTH in the management of acute relapse in MS. PMID- 2544830 TI - [Viral infections and neoplasms of the uterine cervix]. AB - Since the last century, epidemiological studies of cervical carcinoma have shown a close link with sexual activity and in particular with promiscuity starting at an early age. Aetiological research has therefore concentrated on identifying sexually transmitted pathogens. In recent years studies have focused on the apparently significant role of HSV and particularly HPV in the aetiopathogenesis of this tumour. After the first cytohistological findings the HPV-cervical cancer link has been confirmed by electron microscopy, immunohistochemical studies and hybridisation of viral DNA. The identification of different HPV types presenting varying degrees of oncogenic risk offers the prospect of reaching a reliable prognosis on the basis of the particular virus identified in the lesion. The hypothesis that the virus has a decisive influence on the biology of th tumour is also intriguing: findings on the incidence and course of cervical cancer in the youngest women seems to suggest tha HPV may be a fundamental tumour growth factor. PMID- 2544831 TI - Human papillomaviruses and cervical neoplasia: a review. PMID- 2544832 TI - An open study of lisinopril in general practice. PMID- 2544833 TI - Liver disease and hepatitis B infection in a large New Zealand family. AB - This study illustrates the relationship between the hepatitis B virus (HBV) carrier state and primary hepatocellular carcinoma in a large family of Maori (173 members) amongst whom four brothers have died of primary hepatocellular carcinoma. The brothers were from a generation of fourteen siblings, eleven of whom were tested for hepatitis B surface antigen (HBsAg) and all found to be positive. Amongst the offspring of this generation there were 13 HBsAg positives from 28 children (46%) born to female carriers but no HBsAg positives amongst the 28 offspring of male carriers. The study provides further evidence that the morbidity which often follows HBsAg carriage, may be associated with early (perinatal) infection. There was a marked decrease in HBV serologic markers in succeeding generations, from 100% in generation two, to 55% in generation three and 14% in generation four, unrelated to the use of hepatitis B vaccine. PMID- 2544834 TI - ACE inhibitors and anaemia. PMID- 2544835 TI - New tracers for the imaging of the medullary thyroid carcinoma. AB - I-MIBG and 99Tcm(V)-DMSA have been recently proposed as scintigraphic markers of the medullary thyroid carcinoma (MTC). Thirty two patients were examined with 131I- or 123I-MIBG and 26 of these were re-examined with 99Tcm(V)-DMSA (planar and SPECT). From our experience we can draw the following conclusions: (1) the scintigraphic attempt was useless in patients with normal levels of plasma calcitonin (CT): (2) in patients with high plasma CT levels, the sensitivity of the MIBG was better in familiar (3/3 true positive) than in sporadic disease (7/21 true positive; overall sensitivity = 42%); the 99Tcm(V)-DMSA was positive in 16/19 cases, (overall sensitivity = 84%) even in cases which had been false negative with MIBG (6 patients); (3) no false positive results were found; (4) the scan with 99Tcm(V)-DMSA is then suggested as the first imaging approach during the follow up in patients affected by MTC and still having high levels of plasma CT. The MIBG scan should be limited to the patients in whom the possible use of MIBG therapy has to be investigated. PMID- 2544836 TI - Maternal malignancy metastatic to the products of conception: a review. AB - Documented reports of maternal malignancy metastatic to the placenta and fetus are rare. From 1866 until the present there have been 52 cases reported in the Western literature. We report a case of maternal large-cell carcinoma of the lung metastatic to the maternal brain and the placenta without fetal involvement. PMID- 2544837 TI - Combination chemotherapy with methotrexate, adriamycin, cyclophosphamide and CCNU (MACC) for nonsmall cell lung cancer. 4-year experience with 92 patients. AB - Ninety-two nonsmall cell lung cancer (NSCLC) patients were treated with a combination chemotherapy containing methotrexate, adriamycin, cyclophosphamide and CCNU (MACC). The regimen was administered in the dose and schedule originally reported. Median survival for all patients was 32 weeks. Only 6 patients demonstrated an objective response with a median survival rate of 51 weeks. The remaining 70 evaluable patients were nonresponders. These latter patients had a survival probability reduced to 29 weeks. Median time to progression for the whole group was 17 weeks. Partial responses were seen in 3 squamous, 1 large cell carcinoma and 1 adenocarcinoma. One patient with bronchiolo-alveolar carcinoma had complete disease regression and is still alive 136 weeks after starting treatment. Toxicity was significant with 2 treatment-related deaths. The major toxic effects consisted of myelosuppression, nausea, vomiting, and stomatitis. Alopecia was nearly universal; a mild cardiac, renal, or hepatic toxicity was relatively infrequent. Polychemotherapy with MACC regimen may benefit a few selected patients with NSCLC, but its overall antitumor efficacy appears to be very limited. PMID- 2544838 TI - [The spatial-temporal distribution of the mRNA of the Na+-K+-ATPase alpha-subunit in the early development of the clawed toad studied by hybridization in situ]. AB - Distribution of Na+-K+-ATPase alpha-subunit mRNA in full-grown oocytes and early embryos from of X. laevis has been studied using in situ hybridization on histological sections. The mRNA is dispersed about equally in the cytoplasm and is absent from the nucleus. The concentration of the mRNA is successively decreased from fertilization to the late blastula. The drastic decrease in mRNA concentration is observed in a period from morula to mid-blastula. The local increase in the mRNA concentration was detected in dorsal mesoderm and ectoderm during gastrulation. PMID- 2544839 TI - [Characteristics of the electrophysiological parameters of the embryonic cell membranes in the loach during Na+, K+-ATPase inhibition]. AB - Influence of ouabain on changes in transmembrane potential (TMP) and the membrane conductance has been studied in developing embryos of the loach Misgurnus fossilis L. Ouabain does not cause any significant changes in TMP level within 10 15 min after treatment but the membrane was then depolarized to a degree depending on developmental stage. Exposure to ouabain increases the conductance and changes the selectivity of membranes. Reversion potential of ionic current is then decreased from -70- -100 to -5- -30 mV. It is supposed that gradual membrane depolarization owing to the decrease of K+ gradient favors the membrane conductance changes under the influence of ouabain. It has been established that the active transport of Na+ and K+ takes part in realization of the rhythm of TMP level periodical oscillations during synchronous cleavage division. PMID- 2544840 TI - Foscarnet in the treatment of cytomegalovirus retinitis in acquired immune deficiency syndrome. AB - Cytomegalovirus (CMV) retinitis is the major cause of visual loss in acquired immune deficiency syndrome (AIDS). Thirty-one patients with active CMV retinitis were treated with the new antiviral drug, Foscarnet (trisodium phosphonoformate). After a 3-week course of induction therapy, the retinitis improved in 29 of 31 patients (93.5%). Complete resolution of the retinitis was seen in 19 cases (61.3%). Ten patients had partial resolution (32.2%) and two (6.5%) failed to respond. After induction therapy, six patients were put on a low-dose maintenance regimen. All patients without maintenance therapy relapsed within 3 weeks after discontinuation of Foscarnet. The rate of relapse on maintenance therapy was 50% (3/6) within the first 5 weeks. The three other patients of Foscarnet maintenance did not relapse after a follow-up period of 12 weeks. In contrast to ganciclovir, Foscarnet did not induce neutropenia but it produced kidney toxicity that led to reversible renal insufficiency in three cases. Thus, Foscarnet appears to be a useful alternative to ganciclovir, particularly when combined with bone marrow toxic drugs, such as zidovudine (azidothymidine). PMID- 2544842 TI - Congenital hypertrophy of the retinal pigment epithelium in familial adenomatous polyposis. AB - One hundred fifty-three members of 56 kindreds with familial adenomatous polyposis (FAP) underwent funduscopic examination for congenital hypertrophy of the retinal pigment epithelium (CHRPE). All patients underwent wide-angle fundus photography to document lesions, proctosigmoidoscopy to document polyps, and examination for extracolonic manifestations. Ninety-seven patients were diagnosed as having FAP and 56 patients were offspring of FAP patients and thus at 50% risk of inheriting the disease. In two thirds of the kindreds, CHRPE could be used as a congenital phenotypic marker to predict the presence or development of polyps. In these kindreds, all patients with diagnosed FAP and 39% of the patients at risk had at least four CHRPE lesions. In one third of the kindreds, CHRPE could not be used as a predictive congenital marker, and in these kindreds all patients had zero to three total lesions of CHRPE. The presence of CHRPE did not correlate with any other extracolonic manifestations. In kindreds without any other extracolonic manifestations, CHRPE can still be present and can be used as a predictive congenital phenotypic marker. PMID- 2544841 TI - Herpes simplex virus type 1. A cause of the acute retinal necrosis syndrome. AB - The authors have isolated herpes simplex virus type 1 (HSV-1) from the vitreous of two patients with acute retinal necrosis. Clinical and laboratory data suggest that one case represented a primary HSV-1 infection, whereas the other case appeared to be a recurrent HSV-1 infection. In the primary case, changes on magnetic resonance imaging (MRI) suggest spread of the virus posteriorly to both optic tracts and the lateral geniculate ganglia. This case shares many features with the "von Szily" experimental model for HSV retinitis in the mouse. PMID- 2544843 TI - Acyclovir prophylaxis for oral herpes simplex virus infection in patients with bone marrow transplants. AB - Reactivation of herpes simplex virus (HSV) appears to play a significant role in oral mucositis resulting from bone marrow transplantation. The acyclic guanosine derivative acyclovir has been shown to be effective in treating and protecting against HSV infection in this group. The purpose of this study was to determine the role of HSV reactivation in oral mucositis in patients undergoing bone marrow transplantation who were seronegative for HSV or who received acyclovir prophylaxis. The results suggest that HSV plays an insignificant role in oral mucositis in these patients. PMID- 2544844 TI - Ameloblastoma in young persons: a clinicopathologic analysis and etiologic investigation. AB - Ameloblastoma, an odontogenic tumor of ectodermal origin, has been reported to arise, on rare occasions, in a primordial or dentigerous cyst of a young person. Numerous authors have suggested differing nomenclatures for these ameloblastomas (e.g., mural, unicystic, monocystic, intracystic, cystogenic, cystic, plexiform unicystic) and have sought to describe and classify the clinical and histopathologic features. These tumors have been characterized as a distinct variant exhibiting less aggressive behavior and a lower rate of recurrence than conventional ameloblastoma. Furthermore, various etiologic factors have been proposed for these cystic ameloblastomas, including (1) nonspecific irritational factors such as extraction, caries, trauma, infection, inflammation, or tooth eruption; (2) nutritional deficit disorders, and (3) viral infection. The files of the combined accessioned cases of Emory University's and Temple University's oral pathology laboratories were searched and a review of the literature was performed. Thirty-eight cases of mandibular ameloblastoma (37 intraosseous, 1 peripheral) in persons 19-year-old and younger were found from a combined total of 311 accessioned cases of ameloblastoma (12.2%). The average age at diagnosis was 10.4 years for the 18 males and 20 females. Of the 33 cases in which race was stated, 19 (57.6%) were white and 14 (42.4%) were black. In the 28 cases in which a clinical diagnosis was offered, fifteen (53.6%) were thought to be dentigerous cysts. Ten cases from patients less than 19 years old were investigated by means of an immunohistochemical staining technique for the detection of human papilloma virus (HPV) genus-specific structural antigen in formalin-fixed, paraffin embedded tissue. Three of the ten cases (cases 31, 37 and 38) were positive for HPV capsid antigen, whereas none of ten randomly selected ameloblastomas in adults was positive. A discussion of the clinical and histopathologic comparative findings, with emphasis on treatment results and possible HPV etiology, is included. The preliminary nature of finding HPV in the tumor cells is stressed, with recommendation for further verification and typing with the more sensitive in situ hybridization technique. PMID- 2544845 TI - Small round cell tumors. AB - A discussion concentrating on the main tumors composing the round cell tumors of bone. Included topics are Ewing's sarcoma, extranodal lymphoma of bone, primitive neuroectodermal tumor of bone and neuroblastoma metastatic to bone. PMID- 2544846 TI - Fibrous tumors of bone. AB - Benign and malignant fibrous tumors of bone are some of the most common tumors encountered by the orthopedic surgeon. Fibrous dysplasia, fibrous cortical defect, non-ossifying fibroma, benign fibrous histiocytoma and osteofibrous dysplasia are benign tumors best treated conservatively or by intralesional/marginal excision. Desmoplastic fibromas are locally aggressive and require a wide margin for surgical cure. Malignant fibrous histiocytoma and fibrosarcoma of bone are malignant bone tumors and must be treated with wide/radical margins and for malignant fibrous histiocytoma adjunctive chemotherapy. PMID- 2544847 TI - [A gamma scintigraphic assessment of blood supply of the hip joint in Perthes disease]. AB - In order to specify the peculiarities of the blood supply of the hip joint in Perthes' disease the authors carried out gamma scintigraphy by 99m Tc pyrophosphate in 34 children. Patients with roentgenologically confirmed diagnosis of Perthes' disease at various stages as well as those in whom Perthes' disease was suspected, but convincing roentgenologic changes were absent, were examined. The data of scintigraphy and roentgenography were compared. The following phases of disturbances in blood supply were distinguished: ischemia, instable blood supply and stable revascularization. It is supposed that recurrences of ischemia may take place at the stage of resorption. Radionuclide investigation allows to make early diagnosis even before the appearance of roentgenologic changes with differentiation of doubtful cases. PMID- 2544848 TI - Labile repressors are involved in the transcriptional control of PDGF-responsive genes. AB - Platelet-derived growth factor (PDGF) stimulates the transcription of a number of genes in BALB/c-3T3 fibroblasts. Some of these genes (notably the c-myc and c-fo proto-oncogenes) are induced also by phorbol-based tumor promoters which activate protein kinase C. It appears that the response of these genes to PDGF is actually channeled through the activation of protein kinase C. However, other PDGF inducible genes such as JE, KC, and JB do not respond to tumor promoter. Data suggest that a labile repressor protein blocks the transcriptional response of these genes to tumor promoter. This labile repressor is specific for elements in the JE, KC, and JB genes for it has no effect on the activation of the SV40 early promoter, which is a known target for phorbol ester-inducible transativation. PMID- 2544850 TI - Adverse reactions and serologic response to a booster dose of acellular pertussis vaccine in children immunized with acellular or whole-cell vaccine as infants. AB - Adverse reactions and antibody response to pertussis toxin, ie, antitoxin, after a booster injection of an acellular pertussis vaccine were studied in 2-year-old children. A majority (212/241) of the children had previously been immunized with either two or three doses of the acellular vaccine at the ages of 6, 7, and 8 months. The other 29/241 children had received three doses of a plain whole-cell pertussis vaccine at the same ages. In the children who received primary immunization with the acellular vaccine, two cases of more serious systemic reactions occurred in close temporal association with the booster injection. Less serious systemic reactions were few. Local reactions were more common in the children only immunized with the acellular vaccine. Large reactions (greater than 10 cm) were only seen in this group. A good antitoxin response was elicited in both groups. The geometric mean titer in the former whole-cell vaccine recipients was significantly higher than in the children who received only acellular vaccine. The study raised some questions concerning differences in reactogenicity and immune response after a booster injection of an acellular vaccine depending on the type of pertussis vaccine given to infants for the primary immunization. PMID- 2544849 TI - Development of calcium channels in gastric smooth muscle. AB - We used [3H]nitrendipine to characterize dihydropyridine sensitive calcium channels on cells isolated from neonatal (1 d) and weanling (11 wk) rabbit gastric fundic and antral smooth muscle. Incubating with and without nifedipine 20 microM, specific binding was 56 +/- 4% of total binding at 0.1 nM [3H]nitrendipine. Specific binding was saturable, reversible, achieved equilibrium by 10 min at 4 degrees C, and was linearly related to cell concentration. The affinity constant for [3H]nitrendipine was higher in weanling fundus (kd = 243 +/- 121 pM) versus antrum (kd = 771 +/- 190 pM), p less than 0.05. There were no age-related changes in affinity. In the antrum, the number of binding sites (Bmax) increased from 6,000 +/- 266/cell in neonates to 27,500 +/- 8,440/cell in weanlings (p less than 0.05). In the fundus Bmax was 7,750 +/- 2,100/cell in neonates, and there was no age-related change. To assess function, we compared isometric stress in full thickness muscle strips oriented to the circular layer. Bethanechol stimulated dose-dependent tonic contractions in the fundus and phasic contractions in the antrum. Maximal stress increased with age from 305 +/- 54 mN/cm2 to 1140 +/- 73 mN/cm2 (p less than 0.05) in the fundus and from 72 +/- 20 mN/cm2 to 154 +/- 30 mN/cm2 (p less than 0.05) in the antrum. Preincubation and incubation without calcium resulted in reversible inhibition of contraction at both ages. Nifedipine 10-microM inhibited 100% of bethanechol stimulated contraction in the antrum, but only 25% in the fundus at both ages.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544851 TI - [Ulcero-necrotizing enterocolitis: the role of cytomegalovirus. Apropos of a case]. AB - The authors report a case of Wilson-Mikity syndrome in a preterm newborn, that was followed on the 28th day of life by a necrotizing enterocolitis with colic stenosis. The finding of cytomegalic cells on the pathologic examination of the intestinal lesions, the presence of the virus on direct examination of a tissue sample, the elevated IgG levels and the presence of IgM demonstrated a cytomegalovirus infection. The authors discuss the role of this virus in the pathogenesis of the affection, either as directly responsible for the enterocolitis or as a secondary colonization of previous lesions. PMID- 2544853 TI - Effects of ryanodine on the spike after-hyperpolarization in sympathetic neurones of the rat superior cervical ganglion. AB - Effects of ryanodine on sympathetic neurones of the rat superior cervical ganglion were investigated by means of intracellular recording. Ryanodine (1 microM) significantly shortened the after-hyperpolarization (AH) following the spike evoked by current injection or pre-ganglionic stimulation without affecting the configuration of the spikes. The shortening of AH caused by ryanodine was dose-dependent at concentrations between 0.1 and 1 microM and was slowly recovered by washing the tissue over 1 h. A partial inhibition of the apamin sensitive slow component of AH was the maximal effect obtained at 1 microM. Although the input membrane resistance was not changed, ryanodine evoked repetitive discharges at long intervals in response to long depolarizing current pulses applied across the cell membrane. Ryanodine (5 microM) did not depress the Ca-spike but shortened the following AH in a lesser degree than that following the normal spike. Spontaneous small fluctuations of the resting membrane potential were occasionally observed under normal conditions. They were facilitated by caffeine and abolished by ryanodine. Caffeine also enhanced the slow component of the AH but did not affect it in the presence of ryanodine. These results suggest that ryanodine inhibits Ca release from intracellular store sites. The released Ca may contribute to generating the long-lasting AH and to regulating the excitability of rat sympathetic neurones. PMID- 2544852 TI - Kinetics and distribution of voltage-gated Ca, Na and K channels on the somata of rat cerebellar Purkinje cells. AB - Voltage gated ion channels on the somatic membrane of rat cerebellar Purkinje cells were studied in dissociated cell culture with the combination of cell attached and whole-cell variation of patch clamp technique. The method enables us to record local somatic membrane current under an improved space clamp condition. Transient (fast-inactivating) and steady (slow inactivating) Ca channel currents, Na current, transient (fast-inactivating) and steady (slow-inactivating) K currents, were observed. Transient and steady Ca channel currents were activated at test potentials more positive than -40 mV and -20 mV, respectively (in 50 mM external Ba). The transient current inactivated with a half-decay time of 10-30 ms during maintained depolarizing pulses, while the steady current showed relatively little inactivation. Na current was activated at more positive potentials than -60 mV, and inactivated with a half-decay time of less than 5 ms. Transient and steady K outward currents were recorded at more positive potential than -20 mV and -40 mV, respectively. The transient current inactivated with a half-decay time of 2-8 ms. Ca, Na and K channels showed different patterns of distribution on the somatic membrane. Steady Ca channels tended to cluster compared with Na or K channels. PMID- 2544854 TI - Characterization of a novel endonuclease from Crithidia fasciculata. AB - A new endonuclease activity has been identified in whole cell lysates of the trypanosomatid Crithidia fasciculata. This activity, termed endonuclease A (Endo A), introduces single-strand breaks at highly preferred sites in double stranded DNA substrates Physical analysis of this enzyme indicates that it has a sedimentation coefficient S20,W of 4.9 and a Stokes radius of 59A and thus, a native molecular weight of 125,000 and a frictional coefficient of 1.8. A monomeric structure is suggested for the enzyme based on the recovery of Endo A activity associated with a polypeptide with a molecular weight of 116,000 120,000, following electrophoresis on sodium dodecyl sulfate polyacrylamide gels. Endo A shows an absolute requirement for Mg2+ or Mn2+ and exhibits activity over a broad pH and temperature range, with optimal conditions for activity at pH 8.0 and 30 degrees C. PMID- 2544855 TI - Mutant analysis of protein interactions with a nuclear factor I binding site in the SL3-3 virus enhancer. AB - Nuclear factor I (NFI) is shown to be of importance for the activity of the enhancer element of a T-cell leukemogenic murine retrovirus, SL3-3, and for the regulation of this element by glucocorticoid. Each nucleotide of the binding site of the NFI proteins was mutated, and the effects of the mutations were quantitated with an electrophoretic mobility shift assay. Mutations in the inverted repeat of the binding site have symmetric effects which strongly support the notion that NFI proteins preferentially bind to dyad symmetry sites. Such binding sites were shown to be more than 100 fold stronger than the corresponding single binding sites. We find dyad symmetry sequences which are much stronger NFI binding sites than NFI sites identified in different genes and also stronger than previously proposed consensus binding sequences for NFI. PMID- 2544856 TI - Pathways of nucleoprotein assembly on 5S RNA genes in a Xenopus oocyte S-150 extract. AB - Conditions for transcription and nucleosome assembly of plasmids bearing Xenopus 5S RNA genes have been monitored in the whole oocyte S-150 extract (1). We find that the optimal conditions for transcription differ substantially from optimal conditions for nucleosome assembly. DNA molecules bearing as few as 50% of the native density of nucleosomes are transcriptionally inert. Although the 5S gene specific transcription factor TFIIIA is in excess in this extract, these nucleosome reconstitutes do not exhibit TFIIA-like DNase footprints nor do these reconstitutes bind exogenous TFIIIA. We have also examined the nucleotide requirement for DNA supercoiling and for generation of 5S gene transcription complexes. Supercoiling associated with nucleosome assembly does not require ATP; however, nucleotide hydrolysis is required for establishment of active complexes. Phosphorylation of a 200 kdalton protein occurs in a 5S DNA-dependent manner concurrent with the generation of primed transcription complexes. Results of nondenaturing gel electrophoresis coupled with a second dimension of SDS gel electrophoresis suggest that the 200 kD protein may be a component of the 5S RNA gene transcription complex. PMID- 2544858 TI - Nucleotide sequence of cDNA encoding subunit Va from rat heart cytochrome c oxidase. PMID- 2544857 TI - The stereoselective enzymatic synthesis of 9-beta-D-2'-deoxyribofuranosyl 1 deazapurine. AB - The transfer of 2-deoxyribose from thymidine to 1-deazapurine which is catalysed by N-deoxyribosyl transferases from Lactobacillus leichmanii occurs in high yield. This is a very stereoselective process and only one product, 9-beta-D-2' deoxyribofuranosyl 1-deazapurine, is formed. Nmr spectroscopy, and in particular, nuclear Overhauser enhancement experiments, confirm that the 2-deoxyribose moiety is bound to N-9 rather than N-7 and that the glycosidic link has the beta configuration. PMID- 2544859 TI - Nucleotide sequence of cDNA encoding subunit IV of cytochrome c oxidase from fetal rat liver. PMID- 2544860 TI - Nucleotide sequence of the region encompassing the glpKF operon and its upstream region containing a bent DNA sequence of Escherichia coli. PMID- 2544861 TI - Nucleotide sequence of genomic segment 2 of the human rotavirus Wa. PMID- 2544862 TI - Nucleotide sequence of the recA gene of Proteus mirabilis. PMID- 2544863 TI - PCR mediated gene synthesis. PMID- 2544865 TI - [Histopathology of the breast with reference to consultant studies. I. An analysis of 1,339 cases on the breast register of the Fulda Pathologic Institute]. PMID- 2544864 TI - [Inhalation scintigraphy of the lung using the new ultrafine aerosol Technegas]. AB - Evaporation of 99mTc-pertechnetate at about 2500 degrees C on a carbon surface generates an ultrafine aerosol of labelled carbon clusters. Its particle size of about 2-5 nm enables the aerosol to behave similarly to a gas regarding the ability of penetration. After inhalation, the radioactive particles adhere to the walls of the respiratory bronchioli and alveoli. The high concentration of the radioactivity in the argon carrier gas makes it possible to record a scintigram after a single deep breath. We studied four healthy volunteers and 79 patients including 34 with a tentative diagnosis of pulmonary embolism, 20 with bronchus carcinoma, and 15 with various other lung diseases. Ten of these patients were infants, the youngest being nine months old. All patients also had a perfusion scintigraphy. PMID- 2544867 TI - Simultaneous purification of DNA topoisomerase I and II from eukaryotic cells. AB - We have developed a procedure for the simultaneous purification of DNA topoisomerase I and II from calf thymus. Both enzymes were first extracted from isolated nucleoprotein complexes. After batchwise chromatography on hydroxylapatite the two enzyme activities were separated on a FPLC phenylsuperose column. The enzymes were further purified by a second chromatography on phenylsepharose (topo I) or FPLC Mono Q (topo II). The purification can be finished within three days, yielding 0.5-1.0 mg quantities of homogeneous, enzymatic active preparations of the two proteins from 200 g of starting material. PMID- 2544866 TI - [Histopathology of the breast with reference to consultant studies. II. On the diagnosis and differential diagnosis of non-neoplastic diseases and tumors]. PMID- 2544868 TI - [Respiratory complications of inhibitors of conversion enzyme]. PMID- 2544869 TI - [Abnormalities of sodium transport in essential hypertension]. PMID- 2544870 TI - [Meningoencephalitis in Epstein-Barr virus reactivation in a renal transplant patient]. PMID- 2544871 TI - [Cervicovaginal and vulvar papillomavirus lesions: epidemiology in HIV seropositive women. A preliminary study on a continuous series]. AB - In a series of 29 HIV-seropositive women (mean age 27.4 years; 75.9 per cent acquired HIV by intravenous drug abuse) observed over a 2 years' period, we were able to determine the main epidemiological characteristics of cervico-vaginal and vulvar lesions caused by human papillomavirus (HPV). More than 58 per cent of cervico-vaginal smears showed cytological evidence of HPV infection (koilocytosis), and 6 cervical biopsies showed histological abnormalities (from CIN I to invasive carcinoma). Condyloma acuminata was found in 38 per cent of the cases, always associated with cervical HPV lesions. Systematic colposcopy of the whole female genital tract frequently detected multifocal HPV infection. HIV seropositive women constitute a high-risk group for cervico-vaginal and vulvar HPV infection. They clearly need close supervision with frequent cervical smears and, preferably, colposcopy and biopsy for early detection and eradication of genital dysplasias and viral lesions. PMID- 2544872 TI - [Thrombopenia caused by fraxiparin. A case]. PMID- 2544873 TI - [Action of adaptogens: cucurbitacin R diglucoside as a stimulator of arachidonic acid metabolism in the rat adrenal gland]. AB - It has been demonstrated that cucurbitacin R diglucoside (CRD), an adaptogen increasing the rat working capacity and stimulating corticosteroid secretion, stimulates the release of arachidonic acid (AA) in the rat adrenal cortex in vivo (the administration of CRD during 14 days) as well as in vitro (the incubation of isolated rat adrenocortical cells with CRD in the presence of eicosatetraenoic acid, the AA metabolism inhibitor) experiments. The incubation of isolated rat adrenocortical cell with CRD in the presence of AA increases the biosynthesis of 5-HETE, the precursor of which 5-HPETE is known to be a modulator of ACTH-induced corticosteroid secretion. PMID- 2544874 TI - Structure of simian immunodeficiency virus regulatory genes. AB - Three full-length cDNA clones were obtained from cells infected with the simian immunodeficiency virus (SIV) isolated from captive macaques (SIVMAC). Nucleotide sequence analyses suggested that these represented mRNA for the SIV MAC genes tat, rev (formerly, art/trs), and nef (formerly, 3'orf). The putative tat specific clone was active in trans-activation of the SIV MAC long terminal repeat in COS-1 and Jurkat cells. In contrast, the human immunodeficiency virus 1 long terminal repeat was significantly trans-activated only in the COS-1 cells. This suggests that trans-activation by the SIV tat gene is modulated by cell-specific factors. The structure of all of the clones suggested an mRNA splicing pattern more complex than that described for human immunodeficiency virus 1. PMID- 2544875 TI - Transient state kinetic evidence for an oligomer in the mechanism of Na+-H+ exchange. AB - Pre-steady-state kinetic measurements of 22Na+ uptake by the amiloride-sensitive Na+-H+ exchanger in renal brush border membrane vesicles (BBMV) were performed at 0 degrees C to characterize the intermediate reactions of the exchange cycle. At 1 mM Na+, the initial time course of Na+ uptake was resolved into three separate components: (i) a lag phase, (ii) an exponential or "burst" phase, and (iii) a constant velocity or steady-state phase. Pulse-chase experiments using partially loaded BBMV showed no evidence for 22Na+ back-flux, suggesting that the decline in the rate of Na+ uptake rate following the burst represents completion of the first turnover of the exchanger. Gramicidin completely abolished Na+ uptake, indicating that the burst phase results from the translocation of Na+ rather than from residual Na+ binding to external sites. Raising the [Na+] from 1 to 10 mM at constant pH (internal pH 5.7; external pH 7.7) produced a sigmoidal increase in the amplitude of the burst phase without affecting the lag duration or the apparent burst rate. In contrast, Na+ uptake in the steady state obeyed Michaelis Menten kinetics. These results suggest that a minimum of two Na+ transport sites must be occupied to activate Na+ uptake in the pre-steady state. The transition to Michaelis-Menten kinetics in the steady state can be explained by a "flip flop" or alternating site mechanism in which the functional transport unit is an oligomer and only one promoter per cycle is allowed to form a translocation complex with Na+ after the first turnover. PMID- 2544876 TI - Accessibility of receptor-bound urokinase to type-1 plasminogen activator inhibitor. AB - Urokinase plasminogen activator (uPA) interacts with a surface receptor and with specific inhibitors, such as plasminogen activator inhibitor type 1 (PAI-1). These interactions are mediated by two functionally independent domains of the molecule: the catalytic domain (at the carboxyl terminus) and the growth factor domain (at the amino terminus). We have now investigated whether PAI-1 can bind and inhibit receptor-bound uPA. Binding of 125I-labeled ATF (amino-terminal fragment of uPA) to human U937 monocyte-like cells can be competed for by uPA-PAI 1 complexes, but not by PAI-1 alone. Performed 125I-labeled uPA-PAI-1 complexes can bind to uPA receptor with the same binding specificity as uPA. PAI-1 also binds to, and inhibits the activity of, receptor-bound uPA in U937 cells, as shown in U937 cells by a caseinolytic plaque assay. Plasminogen activator activity of these cells is dependent on exogenous uPA, is competed for by receptor-binding diisopropyl fluorophosphate-treated uPA, and is inhibited by the addition of PAI-1. In conclusion, in U937 cells the binding to the receptor does not shield uPA from the action of PAI-1. The possibility that in adherent cells a different localization of PAI-1 and uPA leads to protection of uPA from PAI-1 is to be considered. PMID- 2544877 TI - Specific binding of a HeLa cell nuclear protein to RNA sequences in the human immunodeficiency virus transactivating region. AB - The transactivator protein, tat, encoded by the human immunodeficiency virus is a key regulator of viral transcription. Activation by the tat protein requires sequences downstream of the transcription initiation site called the transactivating region (TAR). RNA derived from the TAR is capable of forming a stable stem-loop structure and the maintenance of both the stem structure and the loop sequences located between +19 and +44 is required for complete in vivo activation by tat. Gel retardation assays with RNA from both wild-type and mutant TAR constructs generated in vitro with SP6 polymerase indicated specific binding of HeLa nuclear proteins to the TAR. To characterize this RNA-protein interaction, a method of chemical "imprinting" has been developed using photoactivated uranyl acetate as the nucleolytic agent. This reagent nicks RNA under physiological conditions at all four nucleotides in a reaction that is independent of sequence and secondary structure. Specific interaction of cellular proteins with TAR RNA could be detected by enhanced cleavages or imprints surrounding the loop region. Mutations that either disrupted stem base-pairing or extensively changed the primary sequence resulted in alterations in the cleavage pattern of the TAR RNA. Structural features of the TAR RNA stem-loop essential for tat activation are also required for specific binding of the HeLa cell nuclear protein. PMID- 2544878 TI - Regulation of transcription by cyclic AMP-dependent protein kinase. AB - cAMP-dependent protein kinase (PKA; ATP: protein phosphotransferase; EC 2.7.1.37) appears to be the major mediator of cAMP responses in mammalian cells. We have investigated the role of PKA subunits in the regulation of specific genes in response to cAMP by cotransfection of wild-type or mutant subunits of PKA together with cAMP-inducible reporter genes. Overexpression of catalytic subunit induced expression from three cAMP-regulated promoters (alpha-subunit, c-fos, E1A) in the absence of elevated levels of cAMP but did not affect expression from two unregulated promoters (Rous sarcoma virus, simian virus 40). Cotransfection of a regulatory subunit gene containing mutations in both cAMP binding sites strongly repressed both basal and induced expression from the cAMP-responsive alpha-subunit promoter without affecting expression from the Rous sarcoma virus promoter. These experiments indicate that cAMP induces gene expression through phosphorylation by the catalytic subunit and that the ambient degree of phosphorylation dictates the level of basal as well as induced expression of the cAMP-regulated alpha-subunit gene. PMID- 2544879 TI - Identification of des-(Gly-Ile)-endozepine as an effector of corticotropin dependent adrenal steroidogenesis: stimulation of cholesterol delivery is mediated by the peripheral benzodiazepine receptor. AB - Delivery of cholesterol to inner mitochondrial membranes is rate-limiting for steroidogenesis in the zona fasciculata of adrenal cortex. A protein that stimulates this process was isolated to homogeneity from bovine adrenal tissue. This protein's primary structure has been determined in its entirety by a combination of automated Edman microsequencing, fast-atom bombardment mass spectrometry (FAB-MS). The sequence was identical to that previously reported for bovine brain endozepine, except that it lacks the last two residues, -Gly-Ile, at the C terminus. To our knowledge, isolation of an endozepine-related protein from a tissue other than brain has not been reported previously. Endozepine competes with benzodiazepines for saturable binding sites in synaptosomes and in mitochondria of specific peripheral tissues. Previous reports have localized the adrenal benzodiazepine receptor to the outer mitochondrial membrane. In this report, we show that the prototypic benzodiazepine, diazepam, effects a stimulation of adrenal mitochondrial cholesterol delivery similar to that observed for endozepine. The effective diazepam concentration was consistent with that previously shown to displace a high-affinity ligand of the mitochondrial benzodiazepine receptor. The action of diazepam in adrenal mitochondria suggests that the mediation of corticotropin-induced steroidogenesis may be the physiological function of the peripheral-type benzodiazepine receptor. These studies provide new insights into the previously unknown function of peripheral benzodiazepine receptors and should allow new investigations into the stimulation of steroidogenesis by endozepines and benzodiazepines in the brain and in certain peripheral tissues. PMID- 2544880 TI - cDNA cloning reveals that the major group rhinovirus receptor on HeLa cells is intercellular adhesion molecule 1. AB - A 90-kDa surface glycoprotein was previously isolated and shown to be required for infection by the "major" group of human rhinovirus (HRV) serotypes. In the present work, the amino acid sequence of the receptor protein was obtained from CNBr and tryptic peptides. Using degenerate oligonucleotides predicted from the peptide sequences, we identified four cDNA clones that encode a 3-kilobase mRNA. The clones were ligated, subcloned in a simian virus 40 expression vector, and used to transfect receptor-negative Vero (monkey) cells. Results showed that transfected cells expressed receptor molecules capable of binding HRV and a monoclonal antibody which recognizes the major group HRV receptor. The cloned receptor cDNA encoded a protein with a sequence nearly identical to that of the intercellular adhesion molecule 1 (ICAM-1), indicating that the two surface proteins are one and the same. Both proteins have identical mass, carbohydrate composition, and tissue distribution. In addition, major group receptors on HeLa cells could be induced with various cytokines in a manner similar to the ICAM-1 ligand. A similar induction of the HRV "minor" group receptor was not observed. PMID- 2544881 TI - cDNA cloning and expression of the human A-type platelet-derived growth factor (PDGF) receptor establishes structural similarity to the B-type PDGF receptor. AB - The primary structure of the human A-type receptor for platelet-derived growth factor (PDGF) has been determined. A 6.5-kilobase (kb) transcript was identified through low-stringency hybridization with a probe derived from the B-type PDGF receptor cDNA. The sequence of a cDNA clone corresponding to the 6.5-kb transcript contains an open reading frame that predicts a 1089-amino acid growth factor receptor-like molecule, which displays 44% overall amino acid similarity with the PDGF B-type receptor. The two receptors have a similar domain organization, with five immunoglobulin-like domains extracellularly and an intracellular split protein tyrosine kinase domain. Transfection of the new cDNA into COS cells led to the expression of a protein specifically recognized by an antiserum previously shown to react with the PDGF A-type receptor. The expressed protein was shown to display high-affinity binding of all three 125I-labeled dimeric forms of PDGF A and B chains in a manner that is characteristic for the PDGF A-type receptor. PMID- 2544882 TI - Expression in bacteria of functional inhibitory subunit of retinal rod cGMP phosphodiesterase. AB - The cGMP phosphodiesterase of vertebrate retinal rod outer segments plays a key role in visual transduction. A functionally active form of the inhibitory gamma subunit of the phosphodiesterase, which keeps the enzyme inactive in the dark, has been obtained in high yield from a synthetic gene expressed in Escherichia coli. A DNA sequence encoding the 87-residue bovine gamma subunit was chemically synthesized and assembled from 10 oligonucleotides. The synthetic gene was cloned into an expression vector that uses the promoter PL of lambda phage. E. coli was transformed with this vector, which encodes a fusion protein consisting of the first 31 residues of the lambda cII protein, a 7-residue joining sequence that is specifically cleaved at its C-terminal end by clotting protease factor Xa, and the 87-residue gamma subunit. The fusion protein was solubilized in 6 M urea and purified by ion-exchange chromatography on a CM-Sephadex column. The typical yield was 1 mg of fusion protein per liter of bacterial culture, which corresponds to the amount of gamma in about 2500 bovine retinas. Proteolytic cleavage of the fusion protein by factor Xa released a synthetic gamma with the same amino acid sequence as that of native gamma. Both fusion protein and synthetic gamma inhibited trypsin-activated phosphodiesterase with high affinity (Kd less than 100 pM). Likewise, both were as effective as native gamma in inhibiting transducin-activated phosphodiesterase in rod outer segment membranes. This inhibition was reversed by the activation of additional transducin. Thus, the N terminus of gamma is not intimately involved in interactions with either the catalytic subunits of the phosphodiesterase or the activated form of transducin. In contrast, a C-terminal deletion mutant terminating at residue 74 of gamma stimulated rather than inhibited the trypsin-activated enzyme. Thus, the C-terminal region of gamma is critical for inhibition of the phosphodiesterase. PMID- 2544883 TI - Iron-sulfur clusters of hydrogenase I and hydrogenase II of Clostridium pasteurianum. AB - The iron and acid-labile sulfide contents and the electron paramagnetic resonance (EPR) properties of hydrogenase I (bidirectional) and hydrogenase II (uptake) of Clostridium pasteurianum (strain W5) have been determined on the basis of quantitative amino acid analyses. The iron and acid-labile sulfide values are approximately 20 and 18 atoms per molecule of hydrogenase I and 14 and 11 atoms per molecule of hydrogenase II, respectively. These amounts are substantially greater than previously reported values, which relied on protein concentration determined by colorimetric assay. The oxidized hydrogenases exhibit unusual EPR signals that originate from a novel type of iron-sulfur center, termed the hydrogenase or H cluster, which covalently binds the inhibitor CO. This EPR signal represents approximately one unpaired electron per molecule in each enzyme with and without bound CO, which is consistent with the presence of one oxidized H cluster (S = 1/2) per enzyme molecule. The two enzymes also contain ferredoxin type four-iron centers or F clusters. The EPR signals from the F clusters observed in the reduced forms of hydrogenase I and hydrogenase II account for approximately four and one unpaired electron per molecule, respectively. We conclude from the iron determinations and the EPR results, together with a reevaluation of previous spectroscopic data, that in both hydrogenases the H cluster probably comprises six iron atoms. Mechanistic models of the two hydrogenases are presented that account for their cluster compositions and the dramatic differences in their catalytic activities. PMID- 2544884 TI - Role of aspartate-96 in proton translocation by bacteriorhodopsin. AB - Proton transfer reactions in bacteriorhodopsin were investigated by Fourier transform infrared spectroscopy, using a mutant protein in which Asp-96 was replaced by Asn-96. By comparison of the BR - K, BR - L, and BR - M difference spectra (BR indicating bacteriorhodopsin ground state and K, L, and M indicating photo-intermediates) of the wild-type protein with the corresponding difference spectra of the mutant protein, detailed insight into the functional role of this residue in the proton pump mechanism is obtained. Asp-96 is protonated in BR, as well as another aspartic residue, which is tentatively assigned to be Asp-115. Asp-96 is not affected in the primary photoreaction. During formation of the L intermediate it is subjected to a change in the H-bonding character of its carboxylic group, but no deprotonation occurs at this reaction step. Also, in the mutant protein a light-induced structural change of the protein interior near the Asn-96 residue is probed. The BR - M difference spectrum of the mutant protein lacks the negative carbonyl band at 1742 cm-1 of Asp-96 and in addition a positive band at about 1378 cm-1, which is most likely to be caused by the carboxylate vibration of Asp-96. This argues for a deprotonation of Asp-96 in the time range of the M intermediate during its photostationary accumulation. On the basis of these results, it is suggested that the point mutation does not induce a gross change of the protein structure, but a proton-binding site in the proton pathway from the cytoplasmic side to the Schiff base is lost. PMID- 2544885 TI - Corticotropin positively regulates its own receptors and cAMP response in cultured bovine adrenal cells. AB - Bovine fasciculata adrenal cells contain specific high-affinity (KD approximately 2.3 +/- 0.4 x 10(-10) M) and low-capacity (1910 +/- 300 sites per cell) corticotropin (ACTH) receptors. Pretreatment of cells with ACTH, caused in a time (maximum effect at 48 hr) and dose-(ED50 approximately 10(-11) M, Vmax = 10(-10) to 10(-9) M) dependent manner an increase in ACTH binding. This was due to a 4 fold increase in the number of binding sites without modification of the binding affinity. The same pretreatment also enhanced the cAMP response to further ACTH stimulation in a dose-dependent manner (ED50 approximately 10(-11) M) and to a lesser extent the response to forskolin. However, pretreatment with higher concentrations of ACTH (10(-8) M) reduced the binding and the cAMP response when compared to the effect of 10(-9) M. These ACTH effects, which were mimicked by 8 bromoadenosine 3',5'-cyclic monophosphate, required de novo protein synthesis. Pretreatment with 10(-13) to 10(-11) M ACTH also enhanced the steroidogenic responsiveness to further hormonal stimulation. However, at higher concentrations the hormone induced an apparent steroidogenic desensitization that was probably related to a depletion of endogenous cholesterol, since cortisol production in the presence of 22-(R)-hydroxycholesterol was increased. Neither angiotensin-II nor atrial natriuretic factor alone modified ACTH receptors, but angiotensin-II partially blocked the stimulatory effect of ACTH. Thus, ACTH is one of the few polypeptide hormones having a positive trophic effect on its own receptors and target-cell responsiveness. PMID- 2544886 TI - An apaH mutation causes AppppA to accumulate and affects motility and catabolite repression in Escherichia coli. AB - apaH- mutants lack the hydrolase responsible for degradation of AppppN dinucleotides in Escherichia coli and show a greater than or equal to 16-fold increase in AppppA under nonstress conditions. These mutants lack detectable activity of sigma F, a factor required for transcription of motility and chemotaxis genes. Expression of the flbB/flaI operon, thought to encode sigma F, is decreased in apaH- mutants, and there appears to be a general decrease in expression of genes regulated by cAMP-binding protein and cAMP as well. PMID- 2544887 TI - Plant retrotransposon from Lilium henryi is related to Ty3 of yeast and the gypsy group of Drosophila. AB - The lily retrotransposon del 1-46 is 9345 base pairs (bp) long. It has long terminal repeats (LTRs) of 2406 bp (left) and 2415 bp (right), which differ in sequence by 1.4%. Sequences similar to those involved in priming DNA synthesis in retroviruses occur in the internal region. Near the left LTR is a sequence complementary to 18 residues at the 3' end of methionine initiator tRNA of three plant species, and a run of 12 purines occurs close to the right LTR. One internal reading frame of del 1-46 has relatively few stop codons. The 1462-codon product from this frame has motifs, in N to C terminus order, corresponding to those identified with RNA binding, protease, reverse transcriptase, RNase H, and integrase functions in retroviruses and certain other retrotransposons. Amino acid sequence comparisons of three conserved pol regions show del to be closely related to the Ty3 retrotransposon of yeast (37-40% identity). del is also related to the gypsy group of Drosophila (17.6, 297, gypsy/mdg4, and 412), showing closer identity with their reverse transcriptase (32-38%) and RNase H (36 45%) domains than with their integrase domain (21-26%). It is proposed that a gypsy group ancestor exchanged the integrase region with a more distantly related element since its divergence from a del/Ty3 common ancestor. The occurrence of related retrotransposons in three different kingdoms (plants, animals, and fungi) strongly implies their horizontal transmission in recent evolutionary time. PMID- 2544888 TI - Specific chromosomal abnormalities characterize fibrosarcomas of bovine papillomavirus type 1 transgenic mice. AB - In the BPV1.69 line of transgenic mice, the bovine papillomavirus type 1 genome elicits both benign dermal fibroblastic proliferation (fibromatoses) and malignant fibrosarcomas. Because these lesions arise only with time, nonviral factors appear to be involved. We have karyotyped several primary tumors as well as a series of low-passage cell lines derived both from fibromatoses and from fibrosarcomas. The fibrosarcomas, but not the preneoplastic fibromatoses, show consistent abnormalities of one or both of two chromosomes, chromosome 8 (trisomy or duplication) and chromosome 14 (monosomy or translocation). The chromosomal abnormalities are not a direct consequence of the viral integration, which we have mapped to chromosome 15 by in situ hybridization. These results suggest that transgenic mice can be used to study the role(s) of cytogenetic changes in tumorigenesis and may direct the search for genes involved in tumor progression. PMID- 2544890 TI - Genomic DNA differences between pathogenic and nonpathogenic Entamoeba histolytica. AB - cDNA libraries were constructed from pathogenic (HM-1:IMSS) and nonpathogenic (SAW 1734) isolates of Entamoeba histolytica. A cDNA clone (cEH-P1) specific for pathogenic amoebae was identified by screening with a pool of sera from patients with invasive amoebiasis that had been absorbed with nonpathogenic amoebae. This clone was used for the identification of a homologous clone (cEH-NP1) in the cDNA from nonpathogenic amoebae. Sequence analysis and comparison of the predicted amino acid sequences for both clones disclosed 12% evolutionary divergence in structure. Hybridization of both cDNA probes to genomic DNA from four pathogenic and five nonpathogenic E. histolytica isolates revealed two distinct Southern blot patterns, one characteristic for pathogenic amoebae and the other for nonpathogenic amoebae. Further, the complex pattern of restriction fragments hybridizing to an actin cDNA probe was also different between pathogenic and nonpathogenic isolates but was conserved within each group of amoebae. The results indicate that pathogenic isolates of E. histolytica are genetically distinct from nonpathogenic isolates. PMID- 2544889 TI - A two-component regulatory system (phoP phoQ) controls Salmonella typhimurium virulence. AB - We have determined that Salmonella typhimurium strains with mutations in the positive regulatory locus phoP are markedly attenuated in virulence for BALB/c mice. The DNA sequence for the phoP locus indicates that it is composed of two genes present in an operon, termed phoP and phoQ. The deduced amino acid sequence of the phoP and phoQ gene products are highly similar to other members of bacterial two-component transcriptional regulators that respond to environmental stimuli. S. typhimurium strains with transposon insertions that create transcriptional and translational gene fusions that require phoP and phoQ for expression have been isolated and have different chromosomal locations, indicating that this system is a regulon. One of these fusion strains, containing a mutation in a gene termed pagC, has a virulence defect. Other strains, including those containing mutations in the phoN gene, encoding an acid phosphatase, have wild-type virulence. Strains with pagC, phoP, or phoQ mutations have decreased survival in cultured mouse macrophages. When used as live vaccines in mice, strains with phoP or phoQ mutations afford partial protection to subsequent challenge by wild-type S. typhimurium. PMID- 2544891 TI - Rate of internalization of an immunotoxin correlates with cytotoxic activity against human tumor cells. AB - The relationship between the cellular internalization of an anti-ganglioside GD2 monoclonal antibody (14.G2a) and the toxic effect of its ricin A-chain immunotoxin (14.G2a-RA) was examined on GD2-bearing M21 human melanoma and T293 small cell lung carcinoma cell lines. The capacity for ligand uptake was determined by examining the parameters that contribute to this constant, including the number of cell-surface binding sites and the internalization rate constant (ke). The maximum uptake of 14.G2a is 11-fold greater for M21 than for T293 cells, due to a 2.7-fold difference in binding sites and a 4-fold difference in the rate of antibody internalization. The capacity for ligand uptake correlates with the cytotoxic activity of the 14.G2a-RA immunotoxin against these two cell lines. Furthermore, we were able to demonstrate that the consequence of internalization of 14.G2a-RA is the intracellular release of undegraded ricin A chain from the antibody. These studies indicate that the rate of internalization is a quantitative parameter that plays a key role in predicting the cytotoxic potency of this immunotoxin. PMID- 2544893 TI - Voltage-gated calcium channels: direct observation of the anomalous mole fraction effect at the single-channel level. AB - Voltage-gated Ca channels are very efficient pores: even while exhibiting strong ionic selectivity, they are highly permeant to divalent cations. Studies of the mechanism of selectivity and ion permeation have demonstrated that whole-cell Ca channel current in mixtures of Ca and Ba ions can be smaller than with equimolar concentrations of either ion alone. This anomalous mole fraction effect (AMFE) has provided an important impetus for proposed mechanisms of ion selectivity and permeation that invoke multiple ion binding sites. However, recordings of unitary L-type Ca currents did not demonstrate the AMFE [Marban, E. & Yue, D.T. (1988) Biophys. J. 55, 594a (abstr.)], raising doubts about whether it is an expression of ion permeation through open Ca channels. We have made patch-clamp recordings from single L-type Ca channels in PC-12 pheochromocytoma cells. Our results demonstrate a significant AMFE at the single-channel level but also indicate that the AMFE can only be found under restrictive conditions of permeant ion concentration and membrane potential. While the AMFE is clear at 0 mV when permeant ions are present at 10 mM, it is not evident when the divalent cation concentration is increased to 110 mM or the membrane potential is hyperpolarized to -40 mV. We compared our experimental observations with predictions of a single file, two-binding-site model of the Ca channel. The model accounts for our experimental results. It predicts an AMFE under conditions that favor ion-ion interactions, as long as the outer binding site is not saturated due to high permeant ion concentration or negative membrane potential. PMID- 2544892 TI - Deltorphins: a family of naturally occurring peptides with high affinity and selectivity for delta opioid binding sites. AB - Deltorphins are endogenous linear heptapeptides, isolated from skin extracts of frogs belonging to the genus Phyllomedusa, that have a higher affinity and selectivity for delta opioid binding sites than any other natural compound known. Two deltorphins with the sequence Tyr-Ala-Phe-Asp(or Glu)-Val-Val-Gly-NH2 have been isolated from skin extracts of Phyllomedusa bicolor. The alanine in position 2 is in the D configuration. These peptides, [D-Ala2]deltorphins I and II, show an even higher affinity for delta receptors than the previously characterized deltorphin, which contains D-methionine as the second amino acid. These peptides show some similarity to another constituent of Phyllomedusa skin, dermorphin, which is highly selective for mu-opioid receptors. These peptides all have the N terminal sequence Tyr-D-Xaa-Phe, where D-Xaa is either D-alanine or D-methionine. While this structure seems to be capable of activating both mu and delta opioid receptors, differences in the C-terminal regions of these peptides are probably responsible for the observed high receptor selectivity of dermorphin and deltorphin. PMID- 2544894 TI - 1,25-Dihydroxyvitamin D3 and the immune system. AB - There is substantial evidence that lymphocytes and monocytes are targets for the actions of the hormonal form of vitamin D, 1,25-(OH)2D3 and that 1,25-(OH)2D3 acts to modulate the proliferation, differentiation, and immune functions of these cells. The effects of the hormone on lymphocytes are mediated directly as well as indirectly via the accessory monocytes. Depending upon the presence or absence of monocytes and the mode of lymphocyte activation, 1,25-(OH)2D3 can either stimulate or suppress lymphocytes. This evidence as well as clinical information and in vivo studies support a role of 1,25-(OH)2D3 in immunobiology. The physiologic, pathophysiologic, and pharmacologic implications of the immunomodulating properties of 1,25-(OH)2D3 however have not been well established. PMID- 2544895 TI - Growth factors in human testis. Ultrastructural demonstration of receptor mediated endocytosis. PMID- 2544896 TI - Mitochondrial encephalomyopathies. PMID- 2544897 TI - Influences of estradiol and of catechol and non-catechol estrogens on the output of prostaglandins in uteri from spayed rats. AB - The effects of 17-beta estradiol and of some catechol and non-catechol-estrogens on the synthesis and output of prostaglandins (PGs) E and F by uteri from ovariectomized rats, were explored. Uteri from castrated animals released twice as much PGE than PGF. When uterine tissue was obtained from spayed rats injected prior to sacrifice with a low dose of 17-beta estradiol (0.5 + 1.0 microgram, on two consecutive days), the output of PGE diminished significantly. With a higher dose of the hormone (0.5 + 50.0 micrograms) the depressive influence on the synthesis and release of PGE was even more marked, whereas the output of PGF rose significantly. Low or high doses of estrone or of estriol failed to affect the release of either one of the PGs determined. On the other hand, 2-0H-estradiol at a low dose had no action but at a higher one inhibited the release of PGE without influencing PGF. Neither low nor high doses of 2-0H estriol or of 2-0H estrone affected the synthesis and release of uterine PGs. It was also observed that all the compounds tested evoked a significant uterotrophic action. It appears plausible that some catechol metabolites of 17-beta estradiol, but not other catechol-estrogens, could be involved in the mechanism of action of 17-beta estradiol modulating the production of PGs by the rat uterus. PMID- 2544898 TI - Radioimmunoassay of LTB4 in plasma from different species: a cautionary note. AB - In clinical and pre-clinical research the pharmacodynamics of selective 5 lipoxygenase and dual 5-lipoxygenase/cyclo-oxygenase inhibitors may be studied by direct RIA of plasma LTB4. Although immunoreactive LTB4 in plasma from A23187 stimulated human blood has the characteristics of authentic LTB4 our results show, particularly in mice and rats, that exposure to A23187 produces large quantities of 12-HETE. Since in different species the levels of 12-HETE increase with platelet concentration we suggest that the 12(S)-HETE is produced by platelet lipoxygenase. However, we do not rule out the possibility that a proportion of 12-HETE may exist as the (R)-stereoisomer. The latter has greater potential for interference in the direct RIA of LTB4. Biosynthesis of 12-HETE may be measured either by RPHPLC/U.V. abs. (8) or by RIA (9) and LTB4 by a more specific antibody described in this report. We conclude that the combined ex vivo RIA of plasma TXB2, LTB4 and 12-HETE has utility in determining the selectivity of inhibitors of arachidonate metabolism and in distinguishing between selective 5-lipoxygenase inhibitors which interact directly with the enzyme and anti oxidant or free radical scavenging types which may be less specific. PMID- 2544899 TI - Population differences in benzodiazepine sensitive male scent-induced analgesia in the deer mouse, Peromyscus maniculatus. AB - We compared opioid and nonopioid involvement in the mediation of scent-induced analgesia in two populations of deer mice, Peromyscus maniculatus; P. m. artemisiae from a mainland region and P. m. angustus from a small marine island. Exposure to bedding taken from the soiled home cage of an isolated (dominant aggressive) male resident elicited a significant increase in the nociceptive responses of male deer mice from mixed sex pairs, with the island population of mice displaying significantly greater analgesic responses than the mainland animals. In the mainland population of mice, the large amplitude analgesia induced by the scent of a conspecific was insensitive to the opiate antagonist, naloxone, but could be blocked by either the benzodiazepine antagonist, Ro 15 1788, or agonist, diazepam. Exposure to the scent of individuals from the island population elicited a lower amplitude analgesia that was sensitive to both the opiate and benzodiazepine manipulations. In the island population, both the lower amplitude analgesia induced by the scent of a conspecific and the higher amplitude analgesic elicited by the scent of a mainland animal was blocked by naloxone and only partially reduced by the benzodiazepine manipulations. Bedding treated with the peppermint also induced analgesia, with the island mice displaying a markedly greater analgesic response than the mainland animals. In both populations of deer mice the peppermint-induced analgesia was blocked by naloxone and insensitive to the benzodiazepine manipulations. These findings are considered in terms of their possible ecological significance and relations to the differences in agonistic and social behaviors between island and mainland populations of deer mice and other small rodents. PMID- 2544900 TI - Anticataleptic effects of the N-methyl-D-aspartate antagonist MK-801 in rats. AB - The N-methyl-D-aspartate (NMDA) antagonist MK-801 was administered to rats in three doses (0.08, 0.16, 0.33 mg/kg) in order to examine its effects on catalepsy that was induced by haloperidol (0.5 mg/kg). The degree of catalepsy was assessed 30 and 60 min after application of drugs by placing the rat on a horizontal bar, on a podium and on a vertical grid. Animals having received saline and haloperidol showed a higher degree of catalepsy than animals having received MK 801 and haloperidol (except for the lowest dose of MK-801). These findings may suggest a therapeutic potential of MK-801 and possibly of other NMDA antagonists in the treatment of Parkinson's disease. PMID- 2544902 TI - Reduction of the intoxicating effects of ethanol by drugs acting at the benzodiazepine-GABA receptor complex. AB - The ability of the benzodiazepine receptor partial inverse agonists Ro 15-4513, Ro 15-3505 and FG 7142, and the picrotoxin site ligands pentylenetetrazole and Ro 5-3663 to reduce ethanol-induced intoxication were investigated. Ro 15-4513 (0.3 3 mg/kg), Ro 15-3505 (3 mg/kg), pentylenetetrazole (20 and 25 mg/kg) and Ro 5 3663 (4 mg/kg) all significantly attenuated the intoxicating effects of ethanol. In contrast, FG 7142 (20 and 40 mg/kg) failed to reduce ethanol intoxication, but reversed the effect of Ro 15-4513. This pattern of results differs from that obtained using other behavioral paradigms. Since drugs which reduce the effects of GABA generally reduce the intoxicating effects of ethanol, it is suggested that the beta-carbolines may be unusual in their interaction with ethanol. PMID- 2544901 TI - Stereochemical effects of 11-OH-delta 8-THC-dimethylheptyl in mice and dogs. AB - The effects of the enantiomers of 11-hydroxy-delta 8-tetrahydrocannabinol dimethylheptyl (11-OH-delta 8-THC-DMH) on spontaneous activity, rectal temperature, tail-flick latency, and catalepsy were studied in mice and in the dog static-ataxia model to determine the relative potency of each enantiomer. The (-)-enantiomer was active in all tests between 3-100 micrograms/kg, while the (+) enantiomer was inactive at 30 mg/kg in the mouse and 1 mg/kg in the dog. The (-) enantiomer was 100-800 times more potent than delta 9-THC in the mouse. The high degree of enantioselectivity and potency are suggestive of an interaction at a specific site such as a receptor. PMID- 2544903 TI - The beta-carboline derivatives ZK 93426 and FG 7142 fail to precipitate abstinence signs in diazepam-dependent cats. AB - The aim of the present study was to investigate the ability of different benzodiazepine recognition site antagonists (Ro 15-1788 and ZK 93426) and inverse agonists (Ro 15-4513, FG 7142 and CGS 8216) to induce abstinence signs in diazepam-dependent cats. Different groups of cats were challenged with each of the benzodiazepine recognition site ligands under investigation 24 hours after the last dose of chronic treatment with diazepam (7 mg/kg, IP at 8.00 a.m. and 8.00 p.m. for 21 consecutive days). The benzodiazepine derivatives Ro 15-4513 and Ro 15-1788 precipitated an abstinence syndrome within minutes after IP administration. The pyrazoloquinoline derivative CGS 8216 also induced withdrawal signs that were less severe and had a longer latency than those elicited by Ro 15 4513 and Ro 15-1788. Abstinence signs included tremors, increased muscle tone, irritability, fear, arched-back posture, pupillary dilation and vocalizations. On the other hand, the beta-carboline derivatives ZK 93426 and FG 7142 failed to precipitate abstinence signs in diazepam-dependent cats when given at doses that prevented the acute effects of diazepam. Our results demonstrate that the ability to induce withdrawal signs in diazepam-dependent cats depends on the chemical structure of the challenge drug (i.e., benzodiazepine or pyrazoloquinoline), since beta-carboline antagonists like ZK 93426 and partial inverse agonists like FG 7142 lack this property. PMID- 2544905 TI - Role of the aromatic group in the inhibition of phencyclidine binding and dopamine uptake by PCP analogs. AB - Thirty-seven arylcyclohexylamines including phencyclidine (PCP) and derivatives, N[1-(2-thienyl)cyclohexyl]piperidine (TCP) and derivatives and N-[1-(2 benzo(b)thiophenyl)cyclohexyl]piperidine (BTCP) were assessed for their ability to inhibit [3H]PCP binding and [3H]dopamine ([3H]DA) synaptosomal uptake. Their pharmacological property (ataxia) was measured by means of the rotarod test. A very good correlation was observed between the inhibition of [3H]PCP binding and the [3H]DA uptake only for arylcyclohexylamines bearing an unmodified phenyl group. Conversely the comparison between the inhibition of [3H]PCP binding and the activity in the rotarod test shows a good correlation with arylcyclohexylamines having any aromatic group (phenyl, substituted phenyl and thienyl rings). This study outlined a new compound (BTCP) without ataxic effect, which is one of the more potent inhibitors of the [3H]DA uptake (IC50 = 8 nM) and which seems very specific since it has a low affinity for [3H]PCP receptors (IC50 = 6 microM). These data show that the aromatic group of the compounds leads to molecules that bind differently to the PCP receptor and to the DA uptake complex. They also suggest that the behavioral properties of arylcyclohexylamines revealed by the rotarod test occur essentially as a result of an interaction with the sites labeled with [3H]PCP and that TCP is more selective than PCP itself in this recognition. PMID- 2544904 TI - Novelty-induced place preference behavior in rats: effects of opiate and dopaminergic drugs. AB - In Experiment 1, adult male rats were given eight 30-min exposures to one of two distinct environments. Control animals received either four exposures to each environment or were not exposed to either environment. When given free-choice access to both environments simultaneously, animals spent significantly more time in the novel environment relative to the familiar environment. In these same animals, horizontal and vertical activity rates were lower in the novel environment than in the familiar environment. In Experiments 2-5, animals were assessed for novelty preference behavior under the influence of either morphine (0, 0.1, 0.3, 1.0 or 3.0 mg/kg), naltrexone (0, 0.1, 0.3 or 1.0 mg/kg), amphetamine (0, 0.1, 0.3 or 1.0 mg/kg) or haloperidol (0, 0.03, 0.1, 0.3 or 1.0 mg/kg). Haloperidol produced a dose-dependent disruption in novelty preference behavior, while all other drugs tested were without effect. Haloperidol also disrupted the novelty-induced decrease in horizontal and vertical activity rates. These results suggest that haloperidol blocks the reinforcing and locomotor depressant effects of a novel environment in a free-choice preference test. PMID- 2544906 TI - Opioid receptor upregulation and supersensitivity in mice: effect of morphine sensitivity. AB - Mice of the Swiss-Webster strain obtained from two suppliers (Taconic, Charles River) were found to differ in their sensitivity to morphine. Mice from Taconic were approximately two-fold more sensitive to the analgesic and lethal effects of morphine compared to the Charles River mice. In a third strain, C3H/HEN, morphine was found to be more than 2.5 times more potent in producing analgesia than in the Charles River mice. Binding studies showed that the Taconic mice and C3H/HEN mice had approximately 40% and 60%, respectively, more specific [3H]naloxone binding sites in brain than did the less sensitive Charles River mice. When treated with chronic naltrexone for 8 days the analgesic potency of morphine was increased by approximately 90% for both Swiss-Webster mice and by 20% for the C3H/HENs. [3H]Naloxone binding was increased by 45-50% in the Swiss-Webster strains, but by only 33% in C3H/HEN mice. These data indicate that receptor upregulation is directly related to increases in morphine potency. Further, these findings suggest that initial sensitivity to morphine can determine the degree of functional supersensitivity and relative receptor upregulation produced by chronic opioid antagonist treatment. PMID- 2544907 TI - Punishment modifies the effects of chlordiazepoxide and benzodiazepine receptors. AB - Littermate groups of male albino rats responded under a procedure which generated comparable rates of punished and nonpunished responding. Chlordiazepoxide (3.0 30.0 mg/kg, IP) increased punished responding but had no effect on nonpunished responding. Homogenate receptor binding studies with [3H]Ro 15-1788 indicated increased benzodiazepine receptor binding in the striatum of rats who received shock. Moreover, a third group of rats exposed to noncontingent shock showed greater increases than those whose responses had been punished, suggesting that predictability and control of shock may have attenuated the effects of the noxious stimulus. Increased binding seen in the cerebellum, however, was related to the punishing effects of the electric shock since it occurred only in those animals receiving response-contingent shock. There were no changes in binding affinity in any of the brain regions tested. Site-specific alterations benzodiazepine receptors following electric footshock are related to the contingencies under which the noxious stimuli are administered. Furthermore, changes in benzodiazepine receptor binding may underlie the differential effects of benzodiazepine agonists on punished and nonpunished responding. PMID- 2544908 TI - Long-term suppression of the cerebral spread of a memory: effects of idazoxan and clonidine. AB - Bitemporal injections of puromycin consistently induce amnesia of aversive maze learning in mice when administered within 3 days of training. These bitemporal puromycin injections lose their amnestic effectiveness if the latency between training and injection is extended beyond 6 days. Consistent with other evidence, we believe that memory (in our task) "spreads" during the 6 days following training. Since previous experiments have indicated that the central noradrenergic system is involved in this process of "memory spread," we have examined the effect of stimulation or blockade of the alpha 2-receptor. To this end, we administered a single dose of the alpha 2-adrenoceptor antagonist, idazoxan, or the alpha 2-agonist, clonidine. Idazoxan (1 mg/kg, SC) had no effect on engram spread. Clonidine (25 micrograms-125 ng/kg, SC), by contrast, suppressed engram spread for at least 30 days after treatment. When mice were tested at 60 and 90 days after treatment, spontaneous recovery (i.e., engram spread) was evident in only about 50% of the clonidine treated mice. Coadministration of idazoxan with clonidine blocked the effects of clonidine on "memory spread." PMID- 2544909 TI - Diazepam reverses the effects of pentylenetetrazole in rat pups by acting at type 2 benzodiazepine receptors. AB - Pentylenetetrazole (75 mg/kg) induced a characteristic coarse body tremor (accompanied by limb extension) and hyperactivity in 4-day-old rat pups. These effects were reversed by diazepam (0.5 and 2 mg/kg) but not by CL 218,872 (10 and 20 mg/kg) which is selective for type 1 benzodiazepine receptors. Diazepam did not affect the brain concentrations of pentylenetrazole, indicating that the reversal was not based on a pharmacokinetic interaction. Neither diazepam nor CL 218,872 had significant effects on the behavior of the rat pups, although diazepam (2 mg/kg) tended to increase locomotor activity. The results suggest that diazepam displays an anticonvulsant effect in the neonatal rat which is mediated by type 2 receptors. PMID- 2544910 TI - In vivo assembly of chromatin on pBR322 sequences cloned into yeast plasmids. AB - In order to study the in vivo assembly of chromatin on prokaryotic DNA templates, we have transformed yeast cells with plasmids pAJ50 and pRB58, which contain pBR322 sequences. In both cases nucleosomes are assembled in vivo on pBR322 DNA, although the nucleosomes are not homogeneous in size. To explore whether there is any preference for nucleosome assembly along pBR322 sequences, we have used an indirect end labeling method. The results indicate that most nucleosomes are placed at random on pBR322, although the probability for histone octamers to interact with some short regions is somewhat reduced. These regions coincide with sequences in which the frequency distribution of nucleosomes reconstituted in vitro (E. Caffarelli et al. (1988) Eur. J. Biochem. 171, 497-501) is low. Sequence determinants that direct chromatin assembly in vitro seem thereby to act to some extent in vivo. PMID- 2544912 TI - Identification of insertion sequence element IS427 in pTiT37 plasmid DNA of an Agrobacterium tumefaciens T37 isolate. AB - The isolation and characterization of an insertion sequence (IS) element, IS427, from Agrobacterium tumefaciens T37 is described. IS427 is present in three nonidentical copies on the pTiT37 plasmid. The copy that was isolated through transposition on the entrapment vector pUCD800 contains at its ends a 16-bp imperfect inverted repeat and generates a 2-bp duplication of the target DNA. IS427 does not show homology with previously characterized IS elements of A. tumefaciens, based on hybridization experiments and/or sequence comparison. PMID- 2544911 TI - ISR1, a transposable DNA sequence resident in Rhizobium class IV strains, shows structural characteristics of classical insertion elements. AB - ISR1 is a small transposable element, identified in Rhizobium class IV strains by its high frequent mutagenic insertion into plasmid RP4. Hybridization studies showed that ISR1 is present in, multiple copies in Rhizobium class IV strains. Nucleotide sequence analysis revealed that ISR1 has a length of 1260 bp and is characterized by perfect inverted repeats of 13 nucleotides followed by a stretch of 28/29 nucleotides with imperfect homology. The insertion under study generated a target site duplication of 4 bp. ISR1 carries a large open reading frame, encoding a putative polypeptide of 278 amino acids (ORFA*), and three smaller ones in antiparallel direction (ORFs A1, A2, A3). Two of them are completely covered by the large open reading frame. No significant homology to 17 other known insertion sequence elements could be detected, either at nucleotide or at amino acid levels. PMID- 2544913 TI - pBR322-derived multicopy plasmids harboring large inserts are often dimers in Escherichia coli K-12. AB - pBR322-related plasmids that are 2.3 to 5.1 kb were found predominantly as monomers, while plasmids that are 7.7 to 15.2 kb were found predominantly as dimers in rec+ cells of Escherichia coli K-12. PMID- 2544914 TI - Plasmid localization and mapping of two Azospirillum brasilense loci that affect exopolysaccharide synthesis. AB - Two Azospirillum brasilense loci that correct Rhizobium meliloti exoB and exoC mutants for exopolysaccharide (EPS) synthesis have been identified previously (K. W. Michiels, J. Vanderleyden, A. P. Van Gool, E. R. Signer, J. Bacteriol., 1988b). A. brasilense exo mutants produce EPS of lower molecular weight than the wild type strain. Here, we show by hybridization that these exo loci are located on a 90-MDa plasmid in A. brasilense Sp7. In four other Azospirillum strains but not in A. lipoferum SpBr17, the loci are likewise located on a plasmid of approximately the same size. Transposon Tn5 insertions in these loci were isolated and mapped on the cloned DNA by restriction analysis. Hybridization of restriction digests of purified 90-MDa plasmid DNA with probes containing the exo loci confirmed their plasmid location. This is the first report on plasmid localization of genes in Azospirillum. PMID- 2544915 TI - Interaction of fucoidan from Pelvetia fastigiata with surface antigens of hepatitis B and woodchuck hepatitis viruses. AB - A sulfated polysaccharide isolated from Pelvetia fastigiata, a marine algae, was found to inhibit in vitro the reaction of the surface antigen of hepatitis B virus (HBsAg) or of woodchuck hepatitis virus (WHsAg) with antibody to HBsAg (anti-HBs). The polysaccharide was composed mainly of 1----2 linked L-fucose-4 sulfate with some (less than 10%) 1----3 linkages. The inhibition of the reaction of HBsAg with anti-HBs or of WHsAg with anti-HBs was found to be directly proportional to the molecular size of the polysaccharide. Comparison of its inhibitory activity with that of carrageenans and dextran sulfates showed that, in addition to the size, the configuration of the component sugar and the presence of deoxy sugar may play a role in the inhibition of reaction of HBsAg or WHsAg with anti-HBs. The fucose sulfate polymer, fucoidan, however, had no effect in vivo on woodchuck hepatitis virus in woodchuck chronic carriers. PMID- 2544916 TI - [The effect of imipramine and trazodone on the re-uptake of (3H) serotonin by thrombocytes in patients with endogenous depression: changes in antidepressive therapy]. AB - The inhibitory potencies of imipramine (IC50-values for IMI) and trazodone (IC50 values for TRA) on platelet [3H]serotonin uptake were measured in depressed patients. The IC50-values for IMI in patients was shown to be higher (P less than 0.01) than in controls. The IC50-values for TRA in patients were lower (P less than 0.01) than found in platelets from controls. These alterations were not accompanied by the significant decrease of a density of platelet [3H]imipramine binding sites. Drug treatment led to the normalization of the IC50-values for IMI and to the partial increase of the IC50-values for TRA. There was a negative correlation of IC50-values for TRA and severity of depressive symptoms evaluated by the Hamilton Depression Rating Scale. The results support the hypothesis that the mechanisms of the regulation of [3H]serotonin uptake sensitivity to IMI and TRA in patients are different. PMID- 2544917 TI - Increased 3H-clonidine binding in the platelets of patients with depressive and schizophrenic disorders. AB - To examine whether alpha 2-adrenergic receptor function is altered in affective and schizophrenic disorders, we determined 3H-clonidine binding in platelets obtained from 33 normal control subjects and from 24 patients with depressive, 22 patients with schizophrenic, 18 with bipolar, and 8 patients with schizoaffective disorders during a drug-free period. The maximum number of binding sites (Bmax) and apparent dissociation constant (Kd) for high affinity 3H-clonidine binding was computed by Scatchard analysis. Comparison of the diagnostic groups indicated that the Bmax in depressed, schizophrenic, and schizoaffective patients was significantly higher than in normal controls, but there were no significant Bmax differences between bipolar patients and controls. Comparison of the Kd among the diagnostic groups indicated no significant differences among the groups or between patient diagnostic groups and normal controls. Baseline Bmax in schizophrenic patients was significantly correlated with the decrease in Brief Psychiatric Rating Scale (BPRS) scores after treatment, suggesting a relationship between baseline Bmax and clinical response. Treatment with lithium caused a significant decrease in the baseline Bmax, whereas treatment with desipramine or trifluoperazine did not cause significant changes in the baseline Bmax. Our results thus indicate an increase in the number of alpha 2-adrenergic receptors in depressed and schizophrenic patients as compared to normal controls. PMID- 2544918 TI - [Differential diagnosis of circumscribed growing breast tumors]. AB - The results of a comparison of clinical, roentgenological and cytological investigation of circumscribed growing tumours of the breast are reported - 43 carcinomas and 19 intracanalicular fibroadenomas - that are hard to differentiate. A number of diagnostically valuable symptoms are described: Significant for carcinoma are an age over 55, microcalcifications (60 +/- 13% of the cases) and different sizes of the tumour found by palpation and mammography (35 +/- 9% of the cases). The diagnostic accuracy of the methods was: clinical 44 +/- 8%, roentgenological 79 +/- 7%, cytological 89 +/- 3% and for their joint application in a special mammographic cabinet 98 +/- 2%. PMID- 2544920 TI - Peritoneal reaction resulting from iodinated contrast material: comparative study. AB - A consensus does not exist as to the optimal contrast agent for hysterosalpingography. This study was undertaken to evaluate the early and delayed inflammatory responses of the peritoneal surfaces to various types of iodinated contrast media. Guinea pigs received intraperitoneal injections of lactated Ringer solution, iothalamate meglumine, diatrizoate sodium, ioxilan, or ethiodized oil. The inflammatory response of the peritoneal surfaces was assessed at 1,7, and 30 days. Five animals were studied at each time point for each agent. No animals that received Ringer lactate or iothalamate meglumine had inflammation at any time. Ioxilan produced inflammation in two of five animals at 7 days and no inflammation at 1 or 30 days. Ethiodized oil produced no inflammation at 1 day; however, three animals had inflammation at 7 days, and all five had inflammation at 30 days. The 30-day group showed striking inflammatory response with granulomatous features. The authors recommend the continued use of meglumine based water-soluble ionic contrast material for hysterosalpingography. PMID- 2544919 TI - Ectopic adrenocorticotropic hormone syndrome: localization studies in 28 patients. AB - A variety of imaging procedures were performed in 28 patients with ectopic adrenocorticotropic hormone (ACTH) syndrome in an attempt to localize the ACTH producing tumor. Diagnosis was made on the basis of removal of an ACTH-producing tumor or biopsy of metastases in the 19 patients with a proved source and the absence of ACTH gradients in bilateral samples of the inferior petrosal sinuses in the nine patients in whom an ACTH-secreting tumor had not been localized. Eleven bronchial carcinoids, two thymic carcinoids, three pheochromocytomas, and three islet-cell tumors constituted the proved sources. The condition has been cured in eight patients, six are alive with residual tumor, and five have died. Of the nine patients with undetected sites of ACTH production, one has died of pneumocystis pneumonia and eight are being treated medically or with bilateral adrenalectomy. Computed tomography (CT) of the chest and abdomen was the most helpful study in the detection of these tumors. Selective arteriography (bronchial and visceral), systemic and portal venous sampling, and iodine-131 meta-iodobenzylguanidine scintigraphy failed to demonstrate tumors when findings at CT were negative. Bronchial carcinoids constituted most of the ACTH-secreting tumors in this study (58%) and in a review of four large series (47%). To assure early detection of these potentially malignant tumors, pulmonary CT should be performed every 6 months, even after hypercortisolism has been medically or surgically controlled. PMID- 2544921 TI - Malignant rhabdoid tumor of the kidney: radiologic features. AB - Malignant rhabdoid tumor of the kidney is a rare neoplasm with distinct clinical and pathologic features. The authors present the radiologic findings of this tumor in two patients. A central soft-tissue mass, compressing and partially obstructing the underlying kidney, in association with a subcapsular fluid collection and capsular involvement, was present in both patients. These features possibly reflect the aggressive nature of malignant rhabdoid tumor. PMID- 2544922 TI - Clinically occult ductal carcinoma in situ detected with mammography: analysis of 100 cases with radiologic-pathologic correlation. AB - One hundred consecutive cases of clinically occult ductal carcinoma in situ (DCIS) detected with mammography were retrospectively analyzed to determine the spectrum of mammographic appearances and to study pathologic correlations. Seventy-two percent of the lesions appeared as microcalcifications, 10% as soft tissue abnormalities, and 12% as a combination of the two. Six percent of lesions were found incidentally in the biopsy specimen. On the basis of mammographic measurements, 22% of the lesions were 5 mm or smaller, and 75% were 20 mm or smaller. Thirty-five percent of the microcalcification clusters were categorized as predominantly casts (linear), 52% as granular, and 13% as granular with several casts. Related pathologic features included the location of the tumor within the ductal system, pattern of growth (histologic subtype), amount and distribution of calcium formation, and presence or absence of reactive changes. Women aged 49 years or less with DCIS were more likely to have microcalcifications and less likely to have a soft-tissue mass than women aged 50 years or more (P = .04). The authors conclude that there is a wide spectrum of mammographic appearances of clinically occult DCIS. PMID- 2544923 TI - Pulmonary infection after cardiac transplantation: clinical and radiologic correlations. AB - Forty-one episodes of radiographically demonstrated pulmonary infection developed in 35 (30%) of 118 cardiac transplant recipients treated with cyclosporine. The most common pathogens were cytomegalovirus (CMV) (13 episodes), Pneumocystis carinii (12 episodes), and Aspergillus (five episodes). Appearance of CMV infection on radiographs was generalized and hazy (n = 9) or limited to one lobe (n = 4). All episodes of P carinii pneumonia, including six combined with CMV infection, appeared diffusely hazy. Aspergillus infection appeared either shaggy and nodular (n = 3) or bibasilar and hazy (n = 2). Aspergillus infection developed only early after transplantation (0.2-2.5 months), whereas CMV infection (1.1-6.1 months) and P carinii pneumonia (2.6-10.3 months) developed later (time ranges for latter two infections exclude three episodes that developed even later in two patients at risk for acquired immunodeficiency syndrome). Nine (8%) patients died of pulmonary infection, eight (7%) within 4.0 months of transplantation. Symptoms or signs were variable; none were found in four (10%) of 41 episodes. The authors recommend frequent chest radiographs in the early months after transplantation. PMID- 2544924 TI - Biliary carcinoma: CT evaluation of extrahepatic spread. AB - The authors studied the computed tomographic (CT) images obtained in 56 patients with pathologically proved biliary cancer and 75 patients with no evidence of biliary disease, attempting to define the normal anatomy of the lymphatic system draining the bile ducts and the prevalence of extrahepatic spread of primary biliary cancer into these retroperitoneal planes. Of 20 patients with gallbladder cancer, 14 (70%) had proved adenopathy and nine (45%) had peritoneal spread at presentation, and another three later developed carcinomatosis. Of 22 patients with proximal cholangiocarcinoma, 16 (73%) had nodal involvement at presentation, four later developed adenopathy, and five had peritoneal dissemination. Distal or diffuse cholangiocarcinomas were less associated with metastatic nodes or peritoneal spread. For all biliary cancers, the nodes most commonly involved were the node of the foramen of Winslow, the superior pancreatoduodenal node, and the posterior pancreatoduodenal chain. Extrahepatic tumor spread produced proximal intestinal obstruction in 13 patients (23%). CT reliably demonstrates lymphatic or other extrahepatic spread of biliary cancers, which may have an important bearing on management decisions. PMID- 2544925 TI - Significance of follicular cyclooxygenase and lipoxygenase pathways of metabolism of arachidonate in sheep. AB - Concurrent changes in concentrations of a product of the cyclooxygenase (prostaglandin [PG] F2 alpha) and lipoxygenase (leukotriene [LT] B4) routes of metabolism of arachidonic acid were measured by radioimmunoassay within the wall of periovulatory ovine follicles. Increased concentrations of PGF2 alpha were detected within follicles before, during and following the time of ovulation. A significant rise in LTB4 was not observed until after follicular rupture had occurred. Inhibition of synthesis of PGF2 alpha by indomethacin was associated with a complete blockade of ovulation. Nordihydroguaiaretic acid, an inhibitor of 5-lipoxygenase, had no effect on ovulation. Periovulatory administration of either drug did not alter sera profiles of progesterone during subsequent luteal phases. These results reconfirm the importance of the cyclooxygenase system in the mechanism of ovulation. It does not appear that follicular LTB4 is a key component in the processes of ovulation or luteinization in sheep. PMID- 2544926 TI - Selective antagonism of peptidoleukotriene responses does not reduce myocardial damage or neutrophil accumulation following coronary artery occlusion with reperfusion. AB - This study was designed to assess the effect of a peptidoleukotriene receptor antagonist, SK&F 104353, for limiting myocardial damage and neutrophil accumulation in rats subjected to myocardial reperfusion injury (MI/R). In conscious rats, SK&F 10,4353 (25 mg/kg, i.v.) antagonized LTD4-induced vasopressor responses by 90% and 60% at 1 and 4 hr, respectively, indicating effective blockade of peptido-leukotriene responses. In another group of animals subjected to 30 min of coronary artery occlusion with reperfusion for 24 hr, myocardial injury and neutrophil infiltration were determined by measuring creatine phosphokinase (CPK) specific activity and myeloperoxidase (MPO) activity, respectively, in the left ventricular free wall (LVFW). Myocardial CPK levels were 8.1 +/- 0.2 U/mg protein in Sham-MI/R vehicle-treated animals, and were significantly decreased to 6.4 +/- 0.6 U/mg protein in MI/R-vehicle animals. Myocardial MPO values were 1.5 +/- 0.5 U/g LVFW in Sham-MI/R vehicle-treated animals, and significantly increased to 4.3 +/- 0.6 U/g LVFW in MI/R-vehicle animals. Administration of SK&F 10,4353 (25 mg/kg, i.v.) 1 min prior to coronary occlusion and 3.5 hr post reperfusion had no effect on the loss of myocardial CPK specific activity or the increase in MPO levels (p greater than 0.05, compared to the MI/R-vehicle group). Thus, at a dose that antagonized LTD4-induced vasopressor responses, SK&F 104353 did not attenuate either the extent of myocardial injury or inflammatory cell accumulation associated with myocardial ischemia/reperfusion. These results suggest that peptidoleukotrienes do not contribute to the progression of myocardial ischemic/reperfusion injury. PMID- 2544927 TI - Effects of prostaglandin E3 and eicosapentaenoic acid on rat bone in organ culture. AB - To assess the possibility that diets rich in eicosapentaenoic acid (EPA) could have adverse effects on the skeleton, we examined the resorptive response to its major project, PGE3, and the effects and metabolism of EPA itself in cultured fetal rat long bones and neonatal rat calvaria. PGE3 stimulated bone resorption with a potency similar to that of PGE2. However, EPA was a much less effective precursor for PGE3 than was arachidonic acid (AA) for PGE2. In bones cultured with complement sufficient rabbit serum, which stimulates endogenous PGE release, addition of EPA had little effect on bone resorption while AA produced a substantial increase. Bones labeled with [3H]-AA and incubated with transforming growth factor-alpha (TGF-alpha), which stimulates endogenous PGE production, produced substantial amounts of PGE2, while bones labeled with [3H]-EPA and treated similarly produced less than 1/10th as much labeled PGE3. Thus, EPA appears to be a less effective precursor for the production of bone resorbing prostanoids than AA in cultured rat bone. However, since PGE3 is a potent stimulator of bone resorption, the possibility that dietary EPA can effect the production of bone resorbing prostanoids in man requires further study. PMID- 2544928 TI - Promotion of differentiation in human colon carcinoma cells by vasoactive intestinal polypeptide. AB - The effects of vasoactive intestinal polypeptide (VIP) and dibutyryl cyclic adenosine 3':5'monophosphate (dbcAMP) on two human colon carcinoma cell lines, HCT 116 and GEO, were investigated. VIP and dbcAMP inhibited the growth of both cell lines in monolayer culture in a dose-dependent manner. Within 6 h of treatment with 1 mM dbcAMP or 0.3 microM VIP, numerous mucin-like droplets were secreted by GEO cells. VIP and dbcAMP also increased carcinoembryonic antigen (CEA) secretion. In both cell lines, a 9-fold increase in conditioned medium CEA levels was observed at 1 mM dbcAMP and a 2.6-fold increase at 1.5 microM VIP. Time- and concentration-dependent evaluation in cAMP levels were elicited by VIP in the two cell lines. Immunocytochemical studies for cell-surface glycoprotein detection in GEO cells showed that VIP induced a morphological and functional organization of mucin-secreting cells. These results indicate that VIP and dbcAMP have antiproliferative and strong differentiation-promoting effects in colon cancer cells. This is the first report of VIP-induced mucin secretion in colon tumor cells. PMID- 2544929 TI - Melanocyte stimulating hormone and melanin concentration hormone may be structurally and evolutionarily related. AB - Two melanotropic peptides, melanin concentration hormone (MCH) and alpha melanocyte stimulating hormone (alpha-MSH), exert opposing actions on melanosome (melanin granule) movements within teleost pigment cells, melanocytes (melanophores). MCH stimulates melanosome aggregation to the cell center whereas alpha-MSH stimulates pigment organelle dispersion out into the dendritic processes of the melanocytes. The actions of alpha-MSH are dependent upon extracellular calcium (Ca2+), whereas those of MCH are actually enhanced in the absence of the cation. At high concentrations (10(-5)-10(-8) M) MCH also exhibits MSH-like activity (autoantagonism), an effect which is abolished in the absence of Ca2+. Therefore, MCH exhibits MCH-like as well as MSH-like activity depending on the presence or absence of extracellular Ca2+. An analogue of MCH, [Ala5, Cys10]MCH, has been synthesized which is totally devoid of MCH activity but still exhibits MSH-like activity. These results suggest that the two melanotropic peptides share some component of structural similarity and may be evolutionarily related. PMID- 2544930 TI - CCK-antagonist L-364,718: influence on rat pancreatic growth induced by caerulein and bombesin-like peptides. AB - The present study investigates the inhibitory effect of the novel potent benzodiazepine-related CCK-antagonist L-364,718 on pancreatic growth in the rat induced by chronic administration of caerulein and bombesin-like peptides. Caerulein, injected s.c. twice daily at a dose of 1 microgram/kg body weight, and bombesin (10 micrograms/kg) induced a similar increase (1.5-3-fold) in pancreatic wet weight, total protein, amylase, trypsin, putrescine and spermidine content after 14 days of treatment. Growth induced by caerulein showed a significant increase in total DNA content suggesting cellular hyperplasia, whereas bombesin like peptides led to cellular hypertrophy. In comparison to bombesin the decapeptide neuromedin C (10 micrograms/kg) was found to be 30-50% less potent. In the same dose range, neuromedin B and the tachykinins neurokinin A and B, all structurally related to bombesin, had no significant trophic effect on the rat pancreas. Administration of the CCK-antagonist L-364,718 twice daily at a dose of 0.1 mg/kg or at 1.0 mg/kg, either s.c. or orally, led dose-dependently to a near complete inhibition of the caerulein-induced trophic effect. In contrast, L 364,718 administered at identical dosages, did not affect pancreatic hypertrophy induced by bombesin and neuromedin C. It is concluded that both peptides mediate their effect on the rat pancreas directly and not via release of endogenous cholecystokinin. Tachykinins are not involved in the regulation of pancreatic growth. Caerulein- and bombesin-like peptides have comparable effects on the stimulation of protein and polyamine synthesis. PMID- 2544931 TI - [Background of low-level gamma-ray spectrometer due to the atmospheric 220Rn and 222Rn]. AB - In connection with low-level gamma-ray counting of natural samples, background due to 220Rn- and 222Rn-daughters was monitored. The results obtained for 4 months showed that the background gamma-ray from 222Rn-daughters was more variable than that of 220Rn-daughters. An efficient air-conditioning was helpful to keep the background stable. It was practically equivalent to the use of N2 gas. Radiometric data for dust samples filtered from the laboratory air suggested that Rn-daughters tended to be removed by air-conditioning. PMID- 2544932 TI - [Nerve paralysis after surgery of the hip. Apropos of 48 cases]. AB - This study is based on 48 cases of paralysis of the femoral or sciatic nerves complicating hip surgery, mostly total hip arthroplasty. Most of these lesions arose during difficult operations. The diagnosis was clinical and electromyographic, but it was not always made from the outset. The prognosis was better for the cases of paralysis of the femoral or peroneal nerves than for the cases of generalised sciatic paralysis in which the recovery, slower as it was, was only satisfactory in 15 cases out of 26. The treatment was primarily medical and required reeducation. A surgical treatment was considered when the electrical exploration showed no improvement after 6 weeks. PMID- 2544933 TI - [Single cutaneous metastasis as the first symptom of a hepatocarcinoma]. PMID- 2544934 TI - [Small cell anaplastic carcinoma of the esophagus]. PMID- 2544936 TI - Complex motion tomography of the sacroiliac joint. An anatomical and roentgenological study. AB - To find a better method for diagnosing sacroiliac (SI) joint disease, an anatomical approach was combined with conventional roentgenology, complex motion tomography and computed tomography. Complex motion tomography is suggested as the method of choice in the investigation of the SI-joint. Because of its complex (sinusoidal) form, the dorsal portion of the joint has to be tomographed in frontal projection and the middle and ventral portions in oblique projection. In 56 patients, referred for probable ankylosing spondylitis, 72 SI joints were investigated. Based on plain radiography six and on frontal tomography five SI joints were diagnosed as normal. However, based on oblique tomography 31 joints were diagnosed as normal. PMID- 2544935 TI - [Improvement in the diagnosis of scrotal diseases using color-coded duplex sonography]. AB - Two hundred and twenty-two patients with abnormalities of their scrotal contents were examined with colour-coded duplex sonography. The aim of this prospective study was to evaluate this new procedure for the investigation of the scrotal organs. Differentiation between testicular torsion and inflammation by colour coded duplex sonography achieved 95% accuracy. The diagnosis of varicoceles was simplified and expedited (accuracy = 99%). In other abnormalities of the scrotum, particularly testicular tumours, a knowledge of the blood flow did not provide any important additional diagnostic information compared with conventional sonography. PMID- 2544937 TI - [Computed tomography of the rotator cuff--a comparison with other imaging technics]. AB - We consider CT to be suitable for investigating the following lesions of the rotator cuff: 1) tendopathia calcarea. 2) Rotator cuff atrophy suitable for surgery. 3) Bone defects. 4) Inflammatory changes with effusions. For demonstrating rotator cuff defects that might require surgery, we recommend CT arthrography, if possible with sagittal sections. This is particularly the case for younger patients with a history of trauma and evidence of a complex shoulder injury and in cases with persistent symptoms after surgical intervention of ineffective conservative treatment. CT, however, is only indicated if sonography supplies limited information or if sonography is negative with positive clinical findings. PMID- 2544938 TI - [Pseudarthrosis of the scaphoid: frequency, pathogenesis and course]. AB - Eight-three scaphoid pseudo-arthroses were found amongst 1.104 scaphoid examinations. Sixty-seven were present at the first examination and 16 pseudoarthroses developed amongst 252 scaphoid fractures. Men were affected predominantly, particularly in the 20 to 40-year old group. Fractures in the proximal third of the scaphoid and vertical oblique fractures had a particular tendency to pseudo-arthrosis formation. The operative treatment of choice is a Matti-Russe bone graft. Only one patient in seven with definite scaphoid pseudo arthrosis showed firm fusion. PMID- 2544939 TI - [Space-occupying lesions in the pineal region--MRT using Gd-DTPA]. AB - The results obtained from contrast-enhanced MRI in 21 patients with masses in the pineal region are reported. The use of gadolinium-DTPA results in intensive contrast uptake by the tumour; this produces better definition of the tumour and more exact delineation from neighbouring structures. This is of importance in judging resectability and in choosing the operative approach. During chemotherapy or radiotherapy, it provides reliable demonstration of the success of treatment and accurately demonstrates the size of the tumour and the presence of necrosis. On the other hand, histologic classification of pineal tumours is possible only occasionally. PMID- 2544940 TI - [The value of magnetic resonance tomography during radiotherapy of childhood tumors in the posterior fossa and the pineal region]. AB - MRT is a highly sensitive method for the diagnosis of childhood tumours in the posterior fossa. Since it demonstrates tumour extent better than does CT, MRT is the method of choice for radiotherapy planning. The result of treatment can be judged morphologically and by measuring relaxation times. Changes due to treatment can be recognised more easily than by CT. A disadvantage of MRT is lack of specificity, since various processes may lead to an equal increase in T1 and T2 times. PMID- 2544941 TI - [Bone marrow changes in Hodgkin's disease: MR tomography and chemical shift imaging]. AB - A patient with Hodgkin's disease, who had received various forms of treatment, was examined by MR tomography and scintigraphy; the results are discussed together with the findings of the bone marrow biopsy. A spectroscopic imaging technique (modified Dixon method) and quantitative evaluation of the fat and water components of the RF signal provide new diagnostic information that may improve the differential diagnosis. Serial examinations following supra-lethal therapy and bone marrow transplantation have been able to show changes in the bone marrow and these could be evaluated quantitatively. PMID- 2544942 TI - [CT morphology and clinical diagnosis of thoracic space-occupying lesions in patients under 40 years of age]. AB - 83 patients with thoracic masses and aged under 40 years, who had been seen in a chest clinic, were examined by CT. They were classified according to their diagnosis and typical CT findings were analysed. Bronchial carcinomas showed predominantly radial extensions and air trapping. Malignant lymphomas and benign tumours usually showed smooth outlines and evidence of some fat content. Malignant mediastinal or neurogenic tumours frequently contained liquid portions and calcification. The CT diagnosis of lymphomas (seven cases), metastases (28 cases) and benign tumours (13 cases) presented no problems in view of the typical appearance and position of the tumours; classification of malignant mediastinal or neurogenic tumours (seven cases) was difficult. Inflammatory masses (14 cases) and bronchial carcinomas (14 cases) showed similar CT appearances. PMID- 2544943 TI - [Lung perfusion-ventilation scintigraphy in squamous cell carcinoma of the lung]. AB - The effect of tumour size, as determined surgically, and tumour position, as determined by bronchoscopy, on ventilation and perfusion scintigraphy was examined in 53 patients with squamous cell carcinomas of the lung. The findings were compared with reduction in one-second-capacity. Central tumours were frequently associated with marked reduction in perfusion. In these patients there was a linear positive correlation between ventilation and one-second-capacity. The reduction in perfusion, and the need to measure this, became with more peripheral tumours. There was a correlation between ventilation and tumour size in patients with V/Q quotient of greater than 1.2. The results show that tumour size and position do not necessarily indicate operability. For planning surgery of central tumours, perfusion scintigraphy therefore occupies an important position. PMID- 2544944 TI - [Follow-up of local lytic therapy of pulmonary emboli using DSA]. AB - The series consisted of 30 patients with pulmonary emboli diagnosed by DSA. In 18 patients repeat DSA was performed using the central catheter which had been employed for the lytic therapy immediately after the end of treatment. Therapy was carried out from two to 14 days. Bleeding was observed in four of the patients. The final DSA showed complete recanalisation in five patients, partial recanalisation in 11 patients and no angiographic improvement in two. PMID- 2544945 TI - [Familial Osler's disease with lung involvement]. AB - Hereditary hemorrhagic teleangiectasia is transmitted by an autosomal dominant character with typical vascular dysplasias in form of teleangiectasias and angiomas which occur on the skin, on the mucous membranes and also in nearly all organs of the body. More than 50% of patients with Osler's disease also have arteriovenous lung malformations. Therefore, pulmonary arteriovenous fistulas should be studied by x-rays for every member of a family suffering from this disease. To prove the suspicion of pulmonary arteriovenous fistula, the procedure of intravenous DSA should be used. PMID- 2544947 TI - [Percutaneous cholecystostomy in treating acute cholecystitis in patients at risk]. AB - Percutaneous cholecystostomy was performed in 17 poor surgical risk patients. 16 patients developed acute acalculous cholecystitis postoperatively in the intensive care unit, 1 patient had an acute cholecystitis with calculi. Diagnostic imaging using CT and US was specific for acute cholecystitis in 58% only. Percutaneous cholecystostomy was the definitive treatment in 69% of the patients. Additional cholecystectomy was required in 3 patients with complicated cholecystitis, in 1 patient with bile leakage after catheter dislocation and in 1 patient with gallbladder calculi. 3 patients died, 2 of them from reasons unrelated to the gallbladder disease. Radiology-guided percutaneous cholecystostomy performed by a transhepatic approach is a safe and effective procedure for acute cholecystitis in high-risk patients. PMID- 2544946 TI - [MRT and CT of diaphragmatic lipomas]. AB - The radiological appearance of 6 posterior epidiaphragmatic fatty tissue masses in MRI and CT is reported. The differential diagnostic considerations of lipoma, herniation or dystopia and the value of imaging techniques are discussed. PMID- 2544948 TI - [Diagnosis of hernia using peritoneography]. AB - Peritoneography was performed in 1200 patients with ill-defined complaints concerning the abdominal wall, the groin or the pelvic floor. The purpose was to exclude or demonstrate the presence of a hernia. Amongst 750 patients, abnormalities were found in 53.5%. The examination was also carried out post operatively in order to demonstrate possible recurrences which were not clinically obvious. Amongst 450 patients, a recurrence or a contra lateral hernia was demonstrated in 44%. These results show that a recurrence can only be demonstrated or excluded with certainty by means of peritoneography. The radiological examination is technically straightforward, can be carried out in a few minutes on an out patient basis and is simple for the patient. PMID- 2544949 TI - Nonocclusive ischaemic bowel disease in patients on chronic haemodialysis. AB - Six patients with chronic renal failure on longstanding haemodialysis are presented, in whom nonocclusive bowel ischaemia occurred as a rare complication of their underlying disorder. Factors implicated in the development of bowel ischaemia in these patients are chronic constipation resulting in increased intraluminal pressure on the bowel wall, premature and progressive arterial disease and bouts of hypotension accompanying the haemodialysis procedure. Contrast studies of the bowel and computed tomography examination can suggest the diagnosis, but angiography alone provides the exact answer in demonstrating nonocclusive mesenteric ischaemia. In patients with chronic renal failure and longstanding dialysis presenting with abdominal symptoms this diagnosis should be considered. PMID- 2544950 TI - Left renal vein compression syndrome ("nutcracker phenomenon"). AB - Four cases are presented with clinical diagnosis of scrotal varicocele on the left side, and one case with ureter varices and left-sided haematuria as a result of compression of the left renal vein between the aorta and superior mesenteric artery (SMA), also known as "nutcracker phenomenon". The clinical signs and the radiological diagnostic methods of the condition are discussed. PMID- 2544951 TI - [Lymphographic and clinical studies following arterial reconstruction in the aortoiliac area and the extremities]. AB - Lymphography was carried out in 41 patients after arterial reconstruction in the aorto-iliac territory or in the extremities: 13 of these patients had been examined pre-operatively. In addition to lymphography, the circumference of the limbs was measured. Lymphographic changes and oedema could be demonstrated in all patients, the extent depending on the type of surgical intervention. There was correlation between the extent of lymphatic damage and the oedema. The duration of post operative oedema depended on effective regeneration of the lymphatics; according to the severity of the surgical injury, regeneration was largely complete from 12 weeks to six months after surgery. PMID- 2544953 TI - [Digital image intensifier radiography--application in angiography, urography and bone diagnosis]. AB - In a retrospective study, the clinical usefulness of digital image intensifier radiography was analysed. It could be demonstrated that for angiographic procedures the image quality of a 1024(2) matrix is superior to a 512(2) matrix. For digital excretory urography it could be seen that in 80 to 90% the images were at least sufficient to analyse the renal pelvis. Digital image intensifier radiography of bones was diagnostically sufficient in 76% and analysis of bone architecture was possible in 85%. PMID- 2544952 TI - [A comparison of digital luminescence radiography and conventional radiodiagnosis in sialography]. AB - In 25 patients sialographies were obtained by both conventional film-screen system combinations and computed radiography (CR), the digital exposure dose being 50% of the conventional one. In CR two differently post processed images were obtained from one x-ray exposure: one "conventionally" processed image resembling a conventional film-screen system combination and a complementary "enhanced" image showing a higher edge enhancement at the same spatial resolution. CR proved to be equivalent to conventional radiography despite the reduction in exposure dose. Using high edge enhancement the peripheral glandular ducts as well as the central ducts superimposed by bony structures were optically accentuated. However, diagnostical information exceeding the conventional film screen system combination was not obtained by CR. Artefacts by post processing procedures relevant for diagnosis were not observed. PMID- 2544954 TI - [A modified laser system for laser angioplasty using a sapphire tip]. AB - This study deals with 26 patients on whom successful laser angioplasty of peripheral vascular occlusions had been carried out, using an Nd-YAG high energy laser system (100 watts) with a sapphire point. The technical requirements are presented and the primary success and complication rates in our material are discussed. PMID- 2544955 TI - [Hemorrhage of the right adrenal gland following liver transplantation]. PMID- 2544956 TI - Haemorrhage in an abdominal cerebrospinal fluid-pseudocyst as a complication of anticoagulant therapy. PMID- 2544957 TI - [Gastrinoma of the duodenal bulb]. PMID- 2544958 TI - [A calcified heterotopic pancreas in the lesser curvature of the stomach]. PMID- 2544959 TI - [Cystitis emphysematosa]. PMID- 2544960 TI - [Ankylosing spondylitis: Andersson lesion]. PMID- 2544961 TI - [A rare hypersupination luxation of the distal radioulnar joint]. PMID- 2544962 TI - [Properties of an orthopoxvirus strain isolated from camels in Niger]. AB - The VD47 viral strain, isolated from camels (Camelus dromedarius) in Niger possesses camelpoxvirus morphology and characteristic properties: heat sensitive, ether resistant, chloroform and IDU sensitive, with ceiling-temperature 38.5 degrees C. It induces formation of syncytial and retracted cell foci with hemadsorption test positive. No pathogen for mice and very mildly for rabbits, this virus is neutralizable by anti-vaccine serum. The question of camelpoxvirus and camelparapoxvirus pathogenicity for human is discussed by the authors. PMID- 2544963 TI - [Metachronous carcinoma of the colon associated with a previous early gastric carcinoma]. PMID- 2544964 TI - [Bronchial cancer with extension into the heart. Apropos of a case with magnetic resonance imaging]. AB - The authors report a case of a 64-year-old patient with an adenocarcinoma of the bronchus with mediastinal involvement. The echocardiography and CT scan showed the existence of involvement of the left auricle. Confirmation and the extent of the intra-cardiac involvement were provided by nuclear magnetic resonance imaging which confirmed the diagnosis here and was particularly helpful. The authors further recall that the place of nuclear magnetic resonance as a standard investigation in assessing staging in bronchial cancer remains to be determined in relation to CT scanning. PMID- 2544965 TI - Regulation of class II major histocompatibility complex genes: relation to multiple sclerosis. PMID- 2544966 TI - Neuroendocrine regulation and the immune response in MS. PMID- 2544967 TI - Endogenous digoxin-like activity of mammalian-lignans and their derivatives. AB - A comparative study was made on the endogenous digoxin-like activity of sixteen mammalian-type lignan derivatives including enterolactone and enterodiol. Cross reactivity to antidigoxin antibody, inhibition of dog kidney Na+,K+-ATPase, and ouabain displacing activity against [3H]ouabain binding to human erythrocytes on the part of these derivatives were examined and compared in all cases with that of ouabain. meso-2,3-Dibenzylbutane-1,4-diol (meso-HA-1) was found to possess the most potent cross-reactivity to antidigoxin antibody. Several lignans, particularly HA-1 (either meso or dl), dl-2,3-bis(3-methoxybenzyl)butane-1,4 diol(HA-4), dl-2,3-bis(3,4-dimethoxybenzyl)butane-1,4-diol(HA-10), dl-2,3-bis(3,4 dimethoxybenzyl)-1,4-dimethoxybutane(HA-11), and trans-2,3-bis(3,4 dimethoxybenzyl)-gamma-butyrolactone(HA-14) were capable of inhibiting Na+,K+ ATPase activity with IC50 at a concentration less than 5 x 10(-4) M. Meso-HA-1, HA-11 and HA-14 also showed [3H]ouabain displacement activity with IC50 at a concentration of 10(-4)-10(-3) M. These determinations of activity were made at doses 100-1000 times those of ouabain. The synthetic lignans are considered to derive in vivo from plant material usually present in fibrous food. Based on the data obtained, some lignans may possibly be endogenous digitalis-like substances. PMID- 2544968 TI - Inhibition by triphenyltin chloride of cytosolic free calcium mobilization in human neutrophils stimulated by a chemotactic factor. AB - We examined the effects of triphenyltin chloride (TPTCl) on the signal transduction in cells, especially the rise of cytosolic free calcium in fura-2 loaded neutrophils stimulated by N-folmyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP). The peak concentration of cytosolic free calcium in neutrophils stimulated by FMLP (10(-7) M) after treatment with 4 mM EGTA was inhibited by TPTCl in a dose-dependent manner and completely blocked in the concentration range of 2.5 to 10 microM in the absence of extracellular calcium. TPTCl also had dose-dependent inhibitory effects on the superoxide anion production and the secretion of beta-glucuronidase in neutrophils stimulated by FMLP (10(-7) M). These results indicate that TPTCl is potent inhibitor of cytosolic free calcium mobilization in FMLP stimulation, associated with superoxide anion production and the secretion of beta-D-glucuronidase. PMID- 2544969 TI - Experimental infection of cattle in early pregnancy with a cytopathic strain of bovine virus diarrhoea virus. AB - Nine pregnant heifers, in early gestation (63 to 107 days), were infected intranasally or in utero with cytopathic bovine virus diarrhoea virus (BVDV) and each dam seroconverted. All nine calves developed to full term; four were stillborn, of which one had seroconverted but virus was not recovered from their tissues. One of the five liveborn calves appeared to have seroconverted in utero to an adventitious BVDV infection in late pregnancy but the remaining four were not viraemic and showed a normal secondary antibody response to BVDV infection at about six months old. Thus, in contrast to results with noncytopathic virus there was no evidence that infection in utero with cytopathic virus could result in a persistent viraemia or immunotolerance. It is suggested that cells able to support a persistent viraemia with cytopathic virus may not be developed in the young fetus. PMID- 2544970 TI - Natural transmission of group A rotavirus within a pig population. AB - Five litters of piglets born within two days of each other, together with their dams, were investigated for faecal excretion of group A rotavirus antigen from birth to two months old. All the 50 piglets in these litters became infected with the virus between 19 and 35 days old. Rotavirus excretion was first seen in one litter which was housed with other litters not included in this study. Two days later, piglets of the second litter in another farrowing room began to excrete rotavirus, and then infection spread to the other three litters in the same room. Within each of these litters, one or two piglets were infected early and thereafter infection spread to other piglets. It took four to 10 days for rotavirus to infect every piglet within a litter, and 16 days in total before all piglets in the five litters were infected. No rotavirus antigen was demonstrated in faeces from sows during the investigation period. PMID- 2544971 TI - Effects of tail docking and castration on behaviour and plasma cortisol concentrations in young lambs. AB - Behavioural and cortisol responses of lambs to the usual husbandry practices of castration and, or, tail docking using tight rubber rings were investigated between birth and seven days old. There were four treatments: control handling and blood sampling (n = 52), tail-docking (T) (n = 57), castration plus tail docking (CT) (n = 54) and intravenous adrenocorticotrophin injection (ACTH) (n = 52). After treatment there was a transient rise in plasma cortisol concentration, the magnitude and duration of which increased in the following order; control, T, CT and ACTH. Behavioural characteristics indicative of distress included restlessness (standing up and lying down frequently, rolling, kicking, stamping), lateral recumbency, immobility with neck extension and hyperventilation. Using changes in cortisol concentrations and behaviour the lamb responses were characterised as reflecting 'mild disturbance without distress' (control, ACTH), 'mild distress' (T) and 'marked distress' (CT). The mild distress lasted for about 30 minutes in T lambs and marked distress for about 60 minutes in CT lambs. PMID- 2544972 TI - Changes in the cortisol responses of lambs to tail docking, castration and ACTH injection during the first seven days after birth. AB - Cortisol responses of Dorset and Scottish Blackface lambs were investigated at six ages between birth and seven days. There were four treatments: control handling and blood sampling (n = 52), tail docking (T) (n = 57), castration plus tail docking (CT) (n = 54), and intravenous adrenocorticotrophin (ACTH) injection (n = 60). Most lambs exhibited transient rises in plasma cortisol concentrations after treatment and the results in individuals were expressed as the area under the cortisol curve (integrated response). Postnatal changes in the integrated responses of control, T and CT lambs differed significantly between the two breeds; in Dorsets they increased markedly between four hours and one day and then decreased, whereas in Scottish Blackface lambs there were no significant changes. The cortisol responses of ACTH lambs decreased markedly between four hours and seven days in both breeds. The results shed light on postnatal changes in the activity of the hypothalamic-pituitary-adrenal axis and suggest for both breeds that the average increments in ACTH secretion caused by noxious stimuli increased markedly during the first one to three days after birth. Whether this reflected parallel increases in the levels of distress experienced by the lambs remains to be clarified. PMID- 2544973 TI - Demonstration of bovine virus diarrhoea virus antigen in formalin fixed, paraffin embedded tissue using a streptavidin/biotin technique. AB - The detection of bovine virus diarrhoea virus (BVDV) antigen in sections from formalin fixed, paraffin embedded tissue is described. Pre-digestion of the sections with 0.02 per cent protease XIV for 18 hours at 4 degrees C is necessary to unmask formalin fixed antigen. A hyperimmune antiserum prepared in a pig, using a combination of BVDV and hog cholera virus inoculations, linked to a biotinylated anti-pig/streptavidin peroxidase detection system demonstrated antigen in a wide range of tissues from cases of mucosal disease and persistently viraemic animals. The inclusion of a monoclonal anti-pig immunoglobulin linked to a biotinylated anti-mouse/streptavidin peroxidase detection system greatly reduced non-specific staining. PMID- 2544974 TI - Ulex europaeus agglutinin 1 lectin histochemical staining of dog hepatocellular and bile duct carcinomas. AB - Ulex europaeus agglutinin 1 (UEA-1) lectin-binding is a feature of canine intrahepatic bile duct epithelial cells but not of canine hepatocytes. In tumours diagnosed histologically as cholangiocarcinomas or hepatocellular carcinomas, UEA 1 staining occurred in tumour cells regardless of histological appearance. The division of primary liver carcinomas into different presumed cell types of origin is not reflected by equivalent differentiation of the tumour cells and is not of diagnostic value. PMID- 2544975 TI - [Atrial natriuretic peptide]. PMID- 2544976 TI - [Obstetrical activity of a regional hospital center in the northern Ivory Coast]. AB - Rupture of the uterus remains, in some countries, a frequent occurrence. This report of the obstetrical activity of a Regional Hospital Center in the Ivory Coast, demonstrates all the difficulties of a disconcerting pathology for visiting physicians, not used to this type of accident in Europe. PMID- 2544977 TI - [Vesicovaginal fistula. Still a daily reality in Africa. Apropos of 89 cases]. AB - Vesico-vaginal fistulae (VVF) are still a daily occurrence in Africa. Obstetrical causes are the most frequent. This severe disability requires that everything be implemented to obtain a leak-free closure of the pathological opening. The authors report their experience with 89 cured cases of VVF. Except for 2 cases cured with an abdominal approach, the Moir technique gave good results in 78 cases, for midline, moderate size, fistulae. In 3 instances, closure of the vagina alone was satisfactory. There were 10 recurrences, cured 9 times at the second operation and once at the third. 3 recurrences came from other institutions and were cured after 4 and 5 successive procedures. In 6 instances, for fistulae close to 5 cm or larger and located in areas difficult to reach, the authors have developed a technique of vaginal closure alone using a turned-over, undetermined vaginal flap creating a leak-free diverticulum. No recurrences have been observed with this technique. PMID- 2544979 TI - Characterization and grouping of Trypanosoma brucei brucei, T.b. gambiense and T.b. rhodesiense by quantitative DNA-cytofluorometry and discriminant analysis. AB - For characterization and differentiation 13 stocks of Trypanosoma brucei spp. were used for a quantitative cytofluorometric determination of their DNA fluorescence intensities by two base-pair-specific fluorochromes. T. b. gambiense could be distinguished from T. b. brucei by an about 20% smaller G-C, and A-T content in the nuclear DNA and a 4% greater G-C and 15% greater A-T content of the kinetoplast DNA. T. b. gambiense could be differentiated from T. b. rhodesiense by a 20% smaller G-C and a 3.8% smaller A-T nuclear DNA content and a 2% greater G-C and a 18% greater A-T DNA content in the kinetoplast. After chromomycin (G-C specific) staining the DNA ratio kinetoplast/nucleus of T. b. gambiense was 6.8, of T. b. brucei 5.2 and of T. b. rhodesiense 5.4. The corresponding values after DAPI (A-T specific) application were 20.8 for T. b. gambiense, 14.4 for T. b. brucei and 16.6 for T. b. rhodesiense. With the fluorescence intensities a discriminant analysis has been computed. After chromomycin application the gambiense stocks could be separated from T. b. brucei and T. b. rhodesiense by this method with a hit rate of 100%. Such perfect separation could not be observed between T. b. brucei and T. b. rhodesiense. Even if most of them were classified into the corresponding subgroups some trypanosomes would nevertheless pass over into the brucei or rhodesiense subgroup and vice versa.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2544978 TI - [Mixed Mullerian tumors. Apropos of a new case with a review of the literature]. AB - The authors report a new case of a Mullerian mixed tumor with malignant double composition with homologous tissue. They are uncommon, probably sometimes unrecognised. These tumors have the same clinical expression but their prognosis is linked to their benign or malignant composition. The treatment proposed by different authors is radical even for benign tumors in the fear of malignant evolution. The number of cases is not sufficient to propose a more economic treatment tested on patients. PMID- 2544981 TI - Beyond ileostomy: surgery for a normal life. PMID- 2544980 TI - [The determination of the geographical boundaries between two subspecies of tsetse flies in the Ivory Coast: Glossina palpalis (Robineau-Desvoidy, 1830) and G.p.gambiensis (Vanderplank, 1949)]. AB - The biometric analysis of the male genitalia of Glossina palpalis palpalis and Glossina palpalis gambiensis about samples caught along four bits of road in Cote d'Ivoire permitted to determine the geographic limit between both subspecies. G. p. palpalis is everywhere in the Guinean area whereas G. p. gambiensis is found in the Sudanese one. In the wet savanna, both subspecies live together and mate to give out a reduced number of hybrids. PMID- 2544982 TI - Morphopathology of toxic neuropathies. PMID- 2544983 TI - Correlation between neutrophil superoxide formation, luminol-augmented chemiluminescence and intracellular Ca2+ levels upon stimulation with leukotriene B4, formylpeptide and phorbolester. AB - When neutrophils were stimulated with leukotriene B4 (LTB4) .O2- -production and elevation of intracellular Ca2+ were initiated more rapidly than after stimulation with N-formyl-methionyl-leucyl-phenylalanine. Phorbol myristate acetate (PMA) induced a substantially more protracted superoxide (.O-2) production. Luminol augmented chemiluminescence (LCL) reflected the kinetic characteristics accurately. The primary distinguishing attribute of LTB4-induced responses were an initial high rate of activation and a rapid termination. Thus, LTB4-induced neutrophil Ca2+-inflow and .O2- -formation may be initiated by mechanisms similar in part to those for formylpeptides; however it does not possess the ability to maintain this activation. The kinetic properties of neutrophil activation may be studied with LCL, irrespective of the exact nature of the LCL-generating oxidants formed by neutrophils. PMID- 2544984 TI - Effect of freezing and ultrasonication on the density of human lymphocyte beta 2 adrenoceptors. AB - In this study 21 pregnant volunteers were divided into three equal groups; blood samples were taken from each, and after isolation of the lymphocytes the cell suspensions were divided into two equal samples. The density of beta 2 adrenoceptor in intact lymphocytes was compared with the receptor density for lymphocytes stored 7 days or 19 days at -70 degrees C and with cells disrupted by ultrasonication. Storage by lymphocytes at -70 degrees C for 7 days to 19 days decreased the beta 2-adrenoceptor density by 20% and 37%, respectively; ultrasonication decreased the density by 65%. PMID- 2544985 TI - Expression of activation markers on CD4+ and CD8+ cells from synovial fluid, synovial tissue, and peripheral blood of patients with inflammatory arthritides. AB - We studied the expression of the Tac antigen, the transferrin receptor (Tfr-R), HLA class II antigens (DR, DQ, DP), CD30, and Act 1 on purified CD4+ and CD8+ cells isolated from synovial fluid (SF), synovial tissue (ST), and peripheral blood (PB) of patients with rheumatoid arthritis (RA) and with non-RA inflammatory arthritides (not ST). Subfractionated T cells of PB from healthy individuals served as controls. SF CD4+ cells from RA and non-RA arthritides expressed the Tac antigen much more frequently than corresponding CD8+ cells (54 and 58% versus 16 and 17%). In contrast, SF CD8+ cells of both patient groups expressed the HLA class II antigens rather more frequently than the corresponding CD4+ cells (88 and 68% versus 72 and 40%). Tfr-R expression was low on CD4+ and CD8+ SF T cells from both patient groups. SF T cells did not express CD30, and their expression of Act 1 did not differ from that of normal PB T cells. The RA ST findings were similar to those of RA SF. The overall expression of activation markers on PB T cells of patients was slightly higher than on those of normal controls, and the RA group was slightly higher than the non-RA group. The results show that intra-articular T cells in arthritis are activated and that CD4+ and CD8+ subsets differ in their expression of Tac antigen and HLA class II antigens. There were also similar patterns of activation markers on both CD4+ and CD8+ SF cells from RA and non-RA arthritis patients, suggesting that several types of arthritis display a similar immunopathogenesis in the joints. PMID- 2544986 TI - The effect of gamma interferon on interleukin 1 release of human monocytes. Dependence on the activating agent and correlation to the mRNA levels. AB - The effect of gamma interferon (IFN-gamma) on interleukin 1 (IL-1) production by human monocytes induced either by bacterial lipopolysaccharide (LPS) or silica dust was examined. Release of biologically active IL-1 from LPS-activated monocytes was enhanced in a dose-dependent manner with IFN-gamma. The augmenting effect of IFN-gamma was most marked in monocytes stimulated with a sub-optimal dose of LPS (1 ng/ml). IFN-gamma delivered the augmenting effect only if present at the beginning of the IL-1 induction. The IL-1 released in the presence of IFN gamma was characterized by gel filtration on fast protein liquid chromatography (FPLC) and by isoelectric focusing; the augmented form of IL-1 was a 17 kDa molecule, with pI 7, i.e. IL-1 beta. In contrast to these data, IFN-gamma had a decreasing effect on IL-1 release by monocytes stimulated with silica dust; in monocyte cultures stimulated with a sub-optimal dose of silica (100 micrograms/ml), only minute amounts of biologically active IL-1 were released in the presence of IFN-gamma. With higher silica concentrations the decreasing effect was less strong. The effects of IFN-gamma on the levels of IL-1 beta mRNA were also analysed. IFN-gamma did not alter the accumulation of IL-1 beta mRNA in LPS-stimulated monocytes. In contrast, in silica-stimulated monocytes IFN-gamma reduced the steady-state levels of IL-1 beta mRNA. Thus, these data indicate that, depending on the stimulating agent, IFN-gamma either down- or up-regulates IL-1 production; the former takes place mainly at the level of transcription, while the latter occurs with post-transcriptional mechanisms. PMID- 2544987 TI - Human CD4-8- -derived clones. Phenotypic and functional characteristics and variation between donors in patterns of T-cell receptor gamma gene rearrangements. AB - Clones were derived from highly purified human CD4-8- lymphocytes from three different donors and maintained in the presence of interleukin 2 and phytohaemagglutinin. Considerable variation was noted between donors in the phenotype and T-cell receptor (TCR) gamma gene rearrangements of CD4-8- -derived clones. In one donor, most clones remained CD4-8- and all were CD3+WT31- and therefore expressed gamma/delta heterodimers. TCR gamma gene rearrangements almost all involved C gamma 1. In contrast, most clones from a second donor were CD3+WT31+, and therefore expressed alpha/beta heterodimers, and many were positive for CD4 or CD8. Most clones from a third donor were CD3+WT31- with a high proportion of TCR gamma gene rearrangements involving C gamma 2. The V gamma 9JP rearrangement was exclusively confined to CD3+WT31- clones and was present in the majority of clones. Almost all CD3+WT31- clones showed TCR beta as well as gamma gene rearrangements. Most CD3+WT31- clones with at least one chromosome rearranged to C gamma 1 exhibited high non-major histocompatibility complex (MHC) restricted cytotoxic activity, while most of those with two C gamma 2 rearrangements, and therefore expressing a non-disulphide-linked gamma/delta heterodimer, had low activity. Preincubation of effector cells with anti-CD3 strongly inhibited the cytotoxicity of CD3+WT31- clones while that of CD3+WT31+ clones was enhanced. This implicates the CD3-gamma/delta complex in target cell recognition by cytotoxic gamma/delta-bearing T-cell clones. The results show that there is heterogeneity between donors in the relative proportions of CD4-8- derived clones expressing alpha/beta heterodimers and the different forms of the gamma/delta heterodimer. PMID- 2544989 TI - Regulation of intercellular adhesion molecule-1 expression on endothelial cells with correlation to lymphocyte-endothelial binding. AB - Intercellular adhesion molecule-1 (ICAM-1) expression on cultured human umbilical cord vein endothelial cells (EC) and its correlation to the leucocyte adherence to EC was investigated. Gamma interferon (IFN-gamma) increases the ICAM-1 expression on EC and concomitantly also increases the binding of leucocytes to EC in vitro. Methylprednisolone down-regulates the IFN-gamma-induced binding, but does not alter the ICAM-1 expression. Leukotriene B4 (LTB4) rapidly increases the binding of lymphocytes to EC, but does not induce ICAM-1 expression on the EC. The dissociation between ICAM-1 expression and leucocyte binding to EC clearly indicates that ICAM-1 is not the only ligand. PMID- 2544988 TI - Differential sensitivities of purified human eosinophils and neutrophils to defined chemotaxins. AB - Functions of eosinophils and neutrophils isolated from normal human blood were determined by measuring chemotactic migration and release of beta-glucuronidase. Four well-characterized chemotaxins, the complement fragment C5a, formyl methionyl-leucyl-phenylalanine (FMLP), platelet-activating factor (PAF), and leukotriene B4 (LTB4) were used as stimuli. Neutrophils showed remarkable chemotactic responses to all four chemotaxins. In contrast, eosinophils showed a significant chemotactic response to C5a and PAF, but only weak responses to FMLP and LTB4. Using these chemotaxins we found the following order of chemotactic potency (maximal number of migrated cells): C5a = LTB4 greater than FMLP greater than PAF for neutrophils and PAF = C5a greater than LTB4 = FMLP for eosinophils. Neutrophils elicited a significant beta-glucuronidase release when stimulated by C5a and FMLP, whereas only small amounts were released with PAF and LTB4. On the other hand, an amount of beta-glucuronidase released from eosinophils comparable to that from neutrophils was elicited only with C5a. FMLP, LTB4, and PAF caused the release of small percentages of beta-glucuronidase. The important cellular functions of eosinophils and neutrophils, chemotaxis and enzyme release, are thought to be controlled by differential responsiveness to stimuli. PMID- 2544990 TI - Microvascular response to locally injected collagenase. An experimental investigation in hamsters and rabbits. AB - Collagenase was injected into the ears of rabbits and the cheek-pouches of hamsters. The acute and long-term microvascular effects were studied by vital microscopy and microangiography. The enzyme was injected at three different concentrations: 120 units/ml, 600 units/ml and 3,000 units/ml. The medium (600 units/ml) and high (3,000 units/ml) concentrations induced effects on the microcirculation such as blood flow impairment and microbleedings. The magnitude of these effects was related to the concentration of the enzyme. Generally, these microvascular effects were of low magnitude as compared with other substances tested using the same experimental models. PMID- 2544991 TI - Immediate silastic arthroplasty for non-salvageable intraarticular phalangeal fractures. AB - Fourteen patients with open non-salvageable intraarticular fractures of the proximal interphalangeal joint or metacarpophalangeal joint, underwent immediate silastic arthroplasty using Swanson H.P. hinged implants. The patients' ages ranged from 14 to 49 years, with an average of 29 years. Time from surgery to final examination ranged from 4 months to 6 years, with an average of 26 months. The active range of motion obtained at the metacarpophalangeal joint arthroplasties ranged from 45 to 80 degrees, with an average of 60 degrees. The active range of motion obtained at the proximal interphalangeal joint arthroplasties ranged from 0 to 70 degrees, with an average of 29 degrees. Better results were generally obtained at the metacarpophalangeal level, and in those arthroplasties performed in non-amputated digits. There were no post-operative infections and all but one joint were stable. PMID- 2544992 TI - [Serologic and virologic studies of selected cattle with antibodies to bovine herpesviruses]. AB - In order to investigate the specificity of low titer antibodies to BHV 1, twelve cattle were subjected to stress and dexamethasone treatment. They were monitored virologically by inoculating cell cultures with naso-pharyngeal-, ocular- and vaginal- or preputial swabs and serologically by assessing the prevalence and incidence of antibodies to bovine, caprine-, porcine-, and equine herpesviruses and to bovine leukemia virus. Antibodies were classified as specific for BHV 1 if the animals excreted IBR virus, or if the antibodies neutralized BHV 1 and reacted with BHV 1 antigens, or if they reacted additionally with CapHV antigens. Animals whose sera recognized BHV 1 and BHV 2 but not other herpesviruses, were judged to have experienced both infections. Nine of the twelve animals had specific BHV 1 antibodies. With three animals the question for specificity of their antibodies remains open. Two animals experienced several herpesvirus infections. Therefore, the induction of crossreacting antibodies, directed against epitopes common to herpesviruses, could not be ruled out. The sera of one animal reacted with BHV 1 and BHV 4 antigens in ELISA tests. They did, however, not neutralize BHV 1. PMID- 2544993 TI - Hydroxyapatite associated disease in an elderly female: differentiation from aseptic necrosis. AB - The crystal arthropathies caused by monosodium urate and calcium pyrophosphate dihydrate are commonly recognised, but the destructive arthropathy associated with the deposition of calcium hydroxy-apatite is less well known in general clinical practice. We describe a case with many of the classical features of hydroxy-apatite associated disease and discuss the features differentiating the condition from aseptic necrosis. PMID- 2544994 TI - Repeat-induced G-C to A-T mutations in Neurospora. AB - In the Neurospora genome duplicate sequences are detected and altered in the sexual phase. Both copies of duplicate genes are inactivated at high frequency, whether or not they are linked. Restriction sites change, and affected sequences typically become heavily methylated. To characterize the alterations of the DNA, duplicated sequences were isolated before and after one or more sexual cycles. DNA sequencing and heteroduplex analyses demonstrated that the process (termed RIP) produces exclusively G-C to A-T mutations. Changes occur principally at sites where adenine is 3' of the changed cytosine. A sequence duplicated at a distant site in the genome lost approximately 10 percent of its G-C pairs in one passage through a cross. A closely linked duplication of the same sequence that was passed twice through a cross lost about half of its G-C pairs. The results suggest a mechanism for the RIP process. PMID- 2544995 TI - Cloning of breakpoints of a chromosome translocation identifies the AN2 locus. AB - Chromosome translocations involving 11p13 have been associated with familial aniridia in two kindreds highlighting the chromosomal localization of the AN2 locus. This locus is also part of the WAGR complex (Wilms tumor, aniridia, genitourinary abnormalities, and mental retardation). In one kindred, the translocation is associated with a deletion, and probes for this region were used to identify and clone the breakpoints of the translocation in the second kindred. Comparison of phage restriction maps exclude the presence of any sizable deletion in this case. Sequences at the chromosome 11 breakpoint are conserved in multiple species, suggesting that the translocation falls within the AN2 gene. PMID- 2544996 TI - Purification and complementary DNA cloning of a receptor for basic fibroblast growth factor. AB - Basic fibroblast growth factor (bFGF) participates in many processes including early developmental events, angiogenesis, wound healing, and maintenance of neuronal cell viability. A 130-kilodalton protein was isolated on the basis of its ability to specifically bind to bFGF. A complementary DNA clone was isolated with an oligonucleotide probe corresponding to determined amino acid sequences of tryptic peptide fragments of the purified protein. The putative bFGF receptor encoded by this complementary DNA is a transmembrane protein that contains three extracellular immunoglobulin-like domains, an unusual acidic region, and an intracellular tyrosine kinase domain. These domains are arranged in a pattern that is different from that of any growth factor receptor described. PMID- 2544997 TI - Human diabetes associated with a deletion of the tyrosine kinase domain of the insulin receptor. AB - The insulin receptor has an intrinsic tyrosine kinase activity that is essential for signal transduction. A mutant insulin receptor gene lacking almost the entire kinase domain has been identified in an individual with type A insulin resistance and acanthosis nigricans. Insulin binding to the erythrocytes or cultured fibroblasts from this individual was normal. However receptor autophosphorylation and tyrosine kinase activity toward an exogenous substrate were reduced in partially purified insulin receptors from the proband's lymphocytes that had been transformed by Epstein-Barr virus. The insulin resistance associated with this mutated gene was inherited by the proband from her mother as an apparently autosomal dominant trait. Thus a deletion in one allele of the insulin receptor gene may be at least partly responsible for some instances of insulin-resistant diabetes. PMID- 2544998 TI - Human diabetes associated with a mutation in the tyrosine kinase domain of the insulin receptor. AB - Insulin receptor complementary DNA has been cloned from an insulin-resistant individual whose receptors have impaired tyrosine protein kinase activity. One of this individual's alleles has a mutation in which valine is substituted for Gly996, the third glycine in the conserved Gly-X-Gly-X-X-Gly motif in the putative binding site fo adenosine triphosphate. Expression of the mutant receptor by transfection into Chinese hamster ovary cells confirmed that the mutation impairs tyrosine kinase activity. PMID- 2544999 TI - Rapid beta-adrenergic modulation of cardiac calcium channel currents by a fast G protein pathway. AB - beta-Adrenergic agonists activate the G protein, Gs, which stimulates cardiac calcium currents by both cytoplasmic, indirect and membrane-delimited, direct pathways. To test whether beta-adrenergic agonists might use both pathways in the heart, isoproterenol was rapidly applied to cardiac myocytes, resulting in a biphasic increase in cardiac calcium channel currents that had time constants of 150 milliseconds and 36 seconds. beta-Adrenergic antagonists of a G protein inhibitor blocked both the fast and slow responses, whereas the adenylyl cyclase activator forskolin produced only the slow response. The presence of a fast pathway in the heart can explain what the slow pathway cannot account for: the ability of cardiac sympathetic nerves to change heart rate within a single beat. PMID- 2545000 TI - [Infantile JRA and ACTH therapy]. AB - Systemic juvenile rheumatoid arthritis is characterized by chronic arthritis and associated with extra-articular features. Although JRA may present at any age throughout childhood, the onset of symptoms during the first year of life is relatively uncommon. The diagnosis of infantile JRA is difficult because of poor complaints. We had a 7-month-old female, first onset at 6 months of age for daily "spiking" fever and skin rash, subsequently developed signs of systemic rheumatoid arthritis. Although aspirin or other nonsteroidal anti-inflammatory agents were used for the therapy, no significant effect was obtained. Prednisone was administered to control arthritis and serious systemic features, however no satisfactory effect was shown. In this patient a single morning alternate-day shot of ACTH-Z (4 units) was employed, and this regimen showed significant effect to control her clinical symptoms. It may be necessary to add a small dose of ACTH Z for control in some cases of infantile JRA patient compared to adult cases. PMID- 2545001 TI - [Catecholamines--therapeutic and adverse effects on the heart]. AB - Catecholamine activation enhances the inotropy of the heart by increasing the sarcolemmal influx of Ca2+. This increased influx is counteracted by an increased sarcolemmal efflux and sarcoplasmic reticulum uptake of Ca2+. Thus the intracellular milieu is protected against a gradual rise in Ca2+ concentration. However, under conditions of continuous, excessive catecholamine release, the heart's potential to remove Ca2+ from the cytosol might become exhausted. This might be caused by a Ca2+-dependent exhaustion of high-energy phosphates. As a result of this, Ca2+ overload of the myocytes and eventually a decrease in the pump function of the heart might occur. This paradox has implications for the clinical management of ischaemic heart disease. PMID- 2545002 TI - Effect of oral acyclovir treatment on symptomatic and asymptomatic virus shedding in recurrent genital herpes. AB - Twenty-six men and women with recurrent genital herpes maintained diaries of their symptoms and signs of infection and submitted 6,515 self-collected cultures during a one-year study of acyclovir therapy. As compared with periods before or after treatment, the mean rates of experiencing symptoms or lesions, and of shedding virus were significantly lower during treatment. Acyclovir treatment reduced the rate of symptomatic shedding from 95 positive cultures to six per 1,000 cultures, but the rate of asymptomatic shedding remained relatively constant, averaging eight per 1,000 cultures. Among the isolates of herpes simplex virus studied, there was no differences in sensitivity to acyclovir between strains recovered on or off therapy or during symptomatic or asymptomatic recurrences. The endonuclease cleavage profiles of asymptomatically shed viruses were essentially the same as those of the symptomatically shed viruses from the same individual. Chronic acyclovir therapy significantly reduced the symptoms and signs of recurrent genital herpes but did not eliminate virus shedding, nor, therefore, the possibility of disease transmission. PMID- 2545004 TI - Vero cells co-infected with Chlamydia trachomatis and herpes simplex virus type 2: a scanning and transmission electron microscope study. AB - Vero (African Green Monkey Kidney) cells infected with Chlamydia trachomatis (serovar L2) (CT-L2) for 48 hr and superinfected with herpes simplex virus type 2 (HSV-2) were fixed 24 hr later and examined with the transmission electron microscope (TEM) and scanning electron microscope (SEM). Ultrastructural examination of the co-infected cells showed that, although many CT-L2 inclusions were present, most were empty of reticulate bodies or elementary bodies. However, large numbers of viruses were observed in the co-infected Vero cells and, in some instances, were seen inside the CT-L2 inclusions. SEM studies of the co-infected cells revealed a swelling and rounding up of the Vero cells typical of a CT-L2 or HSV-2 infection. The outer surface of the co-infected cells were covered with many long microvilli typical of HSV-2 infection with only a few surface protruberances similar to those found in CT-L2 infected cells. PMID- 2545003 TI - In vitro evaluations of condoms with and without nonoxynol 9 as physical and chemical barriers against Chlamydia trachomatis, herpes simplex virus type 2, and human immunodeficiency virus. AB - Simulated in vitro intercourse conditions demonstrated that unlubricated latex condoms provide an effective physical barrier to high concentrations of Chlamydia trachomatis, herpes simplex virus type 2, and human immunodeficiency virus. However, since condoms can be damaged after manufacturing inspection and prior to use, latex condoms alone should not be perceived as absolute protection against STDs. Nonoxynol 9 used in conjunction with condoms provided additional, yet still not foolproof, protection against the three viruses. PMID- 2545006 TI - Implementing universal body substance precautions. PMID- 2545005 TI - Universal precautions: CDC perspective. AB - We know that the risk to health-care workers of acquiring HBV infection in the health-care setting is greater than that of acquiring HIV infection. We know that blood is the primary concern and that the risk following needlestick injury or a cut involving exposure to blood from an HIV-infected individual is approximately 0.5%. We know that transmission can occur without such injuries, but that the risk of such transmission is lower than after a needlestick or cut. We know that many exposures are preventable and that new prevention strategies and technologies are needed. We need to know the risk of infection in groups of health-care workers with extensive blood exposures, whether alterations in the design of devices can reduce the risk of needlesticks and other injuries, and the optimal strategies for ensuring compliance with recommendations. PMID- 2545007 TI - Training of LVNs, housekeeping staff, and other hospital employees. PMID- 2545008 TI - [Malignant diseases of the inner nose--epidemiology and occupational medicine aspects]. AB - Squamous cell carcinomas are the most frequent malignancies of the inner nose, followed by adenocarcinomas, adenoid cystic carcinomas, and other malignant neoplasms. Carcinomas of the nose can be recognized as occupational diseases if there has been a professional exposition to ionizing rays, certain arsenic compounds, hexavalent chrome compounds, nickel, oak or beech wood dust. The sources of danger relevant in industrial medicine are indicated. At present, adenocarcinomas induced by dust of wood are of special significance: 16 out of 22 carcinomas of the nose recognized as occupational diseases between 1978 and 1986 are due to oak and beech wood dust. PMID- 2545009 TI - [Manifestations and therapy of malignant fibrous histiocytomas of the head and neck]. AB - From 1982 through 1987, ten patients with malignant fibrous histiocytomas (MFH) in the head and neck area were treated at the ORL Department of the Central Hospital of Bremen. The primary tumor was situated in the region of the paranasal sinuses in six cases and in the parotid gland, the thyroid gland, the inferior maxilla, and the petrosal bone in one case, each. Nine patients were submitted to primary surgery. A postoperative irradiation was performed in four cases, and in four further cases this was done only when a recurrent tumor had been demonstrated. Three patients received a combined chemotherapeutical treatment. Eight patients died, the median survival time was 15 months. Besides local recurrences which occurred frequently, remote metastases are the decisive factor for the further evolution of the disease. The lung was the most common site of metastatic spread. Only two patients with MFH of the paranasal sinuses are living now without recurrence after an observation time of 24 and 36 months, respectively. PMID- 2545010 TI - [Radiotherapy results in malignant tumors of the inner nose and paranasal sinuses with special reference to neutron therapy]. AB - Radiotherapy in advanced and highly differentiated tumors of the inner nose and the paranasal sinuses is a particular challenge. A report is given about indication, performance and results of neutron therapy in 16 patients treated between 1976 and 1987 at the Radiological University Hospital Hamburg-Eppendorf. Neutron therapy proves to be of great efficiency, especially in case of adenoid cystic carcinomas as well as highly differentiated adenocarcinomas and sarcomas. PMID- 2545011 TI - Adverse effects of preoperative hepatic artery chemoembolization for resectable hepatocellular carcinoma: a retrospective comparison of 138 liver resections. AB - During the past 8 years we have treated 268 patients with primary hepatocellular carcinoma (HCC); total extirpation of the tumor was carried out in 138. Thirty one of the patients with resectable HCCs had already been treated with transcatheter arterial embolization (TAE) of the liver before they were referred to us. The clinical values of preoperative TAE were retrospectively evaluated for those 31 patients and for the remaining 107 patients without TAE. There were no substantial differences between the two study groups in the clinical and histopathologic backgrounds. No differences were observed in the extent of liver resection, estimated blood loss during surgery, and operation time. During surgery, however, troublesome intra-abdominal complications relevant to TAE were encountered in 15 patients, and detection of tumors was impossible, even with intraoperative ultrasonography, in five patients in the group with TAE. Such findings were not present in any of the patients without TAE. Postoperative morbidity and mortality rates were similar in the two groups. There was no significant difference in the rate of recurrence of tumor in the liver, but the recurrence time was significantly shorter in the group with TAE. TAE did not improve the long-term survival rates in patients either with or without cirrhosis. Results of our study seem to indicate that preoperative TAE is meaningless in the treatment of resectable HCCs and therefore should be avoided, particularly in patients with advanced cirrhosis of the liver. PMID- 2545013 TI - [Acute respiratory viral infections]. PMID- 2545012 TI - Presence of calcium/calmodulin-dependent protein kinase II in nerve terminals of rat brain. AB - Calcium/calmodulin-dependent protein kinase type II, a multimeric 550-650 kilodalton enzyme composed of major alpha (50 kilodalton) and beta/beta' (60/58 kilodalton) subunits, is present in high concentrations in mammalian brain. Previous immunocytochemical studies indicated that the enzyme is enriched in cell bodies and dendrites, but did not show a clear-cut localization in nerve terminals. The present study presents evidence, using lesion-induced degenerations of pre- and postsynaptic neuronal populations in the neostriatum and substantia nigra, that calcium/calmodulin-dependent protein kinase II, as measured both by autophosphorylation of enzyme subunits and by synapsin I kinase activity, is present in high concentrations in several populations of presynaptic terminals. Lesions of the corticostriatal tract decreased the amount of enzyme by 30-40% in the neostriatum, a decrease similar to that seen in the same region of synapsin I, a general nerve terminal marker. Lesions of the striatonigral tract induced an even more pronounced decrease of the enzyme in the substantia nigra; this decrease was larger than the lesion-induced change of synapsin I seen in the same region. Our data therefore indicate that certain nerve terminal populations in the rat brain contain high levels of calcium/calmodulin-dependent protein kinase II. PMID- 2545014 TI - [Pathogenesis of acute pneumonia]. AB - The patients with acute pneumonias demonstrated interdependent changes in lipid peroxidation (LPO), antioxidant system (AOS), immune system (IS), and in the pituitary-adrenocortical system (PAS), related to the character of the disease course. The most pronounced changes were seen in the patients with acute pneumonias eventuating in pneumofibrosis. The high level of LPO was combined with AOS depletion, immunodeficiency formation, and with dysfunction of the PAS. Antibacterial treatment did not exert any appreciable effect on the characteristics under study. Thus, the level of LPO and AOS status are important components in the pathogenesis of acute pneumonias, determining the character of the disease course and outcome. It is advisable that research work aimed at the design of the principles of antioxidant therapy may be intensified. PMID- 2545015 TI - [Sphingomyelinase activity in a patient with Niemann-Pick disease]. AB - In a few months old baby a visceromegaly was found, without other clinical abnormalities. Using an artificial substrate a deficiency of sphingomyelinase could not be demonstrated. Histopathological examination of biopsied liver revealed a lipid storage disease. Using the natural substrate the deficiency of sphingomyelinase could be demonstrated in leucocytes and in cultured fibroblasts. PMID- 2545017 TI - Inhibition of hydroxyl-radical-generated DNA degradation by metallothionein. AB - The effect of metallothionein (MT) on hydroxyl-radical-induced DNA damage was investigated in an in-vitro study. The degradation of DNA as demonstrated by agarose gel electrophoresis was inhibited by MT in a concentration-dependent manner. The degradation of DNA was almost completely inhibited by a concentration of 13 microM MT, whereas a concentration of 10 mM glutathione was needed to achieve the same protective effect. PMID- 2545016 TI - Acute and chronic effects of the pesticide amitraz on alpha 2-adrenoceptors in mouse brain. AB - There is increasing evidence to suggest that several effects of the formamidine pesticide amitraz (AMZ) in mammals are mediated by its interaction with alpha 2 adrenoceptors. AMZ has been shown to inhibit the binding of [3H]clonidine, a specific ligand for alpha 2-adrenoceptors to mouse brain in vitro and after administration in vivo. In the present study we have further investigated and characterized the effects of acute and chronic administration of AMZ on brain alpha 2-adrenoceptors in mice. AMZ caused a dose-dependent inhibition of [3H]clonidine binding. This inhibition was long-lasting (more than 48 h) following a relatively high dose of AMZ (75 mg/kg), while it was of short duration (2 h) following low doses (7.5 and 12.5 mg/kg). The time course of inhibition of [3H]clonidine binding was correlated with the plasma levels of AMZ and/or its active metabolites, measured with a novel radioreceptor binding technique. The alteration of [3H]clonidine binding was due to a decrease in alpha 2-adrenoceptor affinity, with no change in the density of binding sites, and was reversible in vitro upon repeated washing of the membrane preparation. Repeated administration of 7.5 mg/kg or 12.5 mg/kg AMZ, to yield a total dose of 75 mg/kg, showed no evidence of a cumulative effect on brain alpha 2-adrenoceptors. PMID- 2545018 TI - Successful treatment of severe cytomegalovirus infections with ganciclovir and CMV hyperimmune globulin in liver transplant recipients. PMID- 2545019 TI - The impact of cyclosporine on the practice of renal transplantation. PMID- 2545020 TI - VP16, epirubicin and procarbazine in the treatment of advanced non-small-cell lung cancer. AB - We report the results obtained in the treatment of 52 advanced non-small-cell lung cancer patients with the combination chemotherapy VP16 (120 mg/m2 i.v., days 1-2-3), epirubicin (50 mg/m2 i.v., day 1) and procarbazine (100 mg/m2 p.o., days 1 through 8). The courses were repeated every 21 days. No patient had been pretreated. A median of 5 courses was administered. Partial response was obtained in 33% and no change in 21% of patients. Median remission time was 6.5 months, and median survival of responders was 10 months. The best response rate and median survival were obtained in the lowest grade performance status patients and in locally advanced disease patients. Major chemotherapy related toxicities were grade 1-2 leukopenia and grade 2-3 alopecia. PMID- 2545022 TI - [Preparation and properties of immobilized riboflavin kinase from Pichia guilliermondii]. AB - Riboflavin kinase (ATP: riboflavin-5'-phosphotransferase, EC 2.7.1.26) from n alkane utilizing Pichia guilliermondii yeast has been immobilized by covalent attachment to CNBr-activated agarose beads. The enzyme activity yield during immobilization reached 71.6%. Immobilized riboflavin kinase showed no significant changes in temperature and pH optima as well as in specificity of the action in relation to synthetic substrate analogues with the substitution of methyl groups at positions 7 and 8 of the isoalloxazine ring. Immobilized riboflavin kinase was stable during FMN synthesis in the continuous-flow packed column enzyme reactor with half-life of 27 days. PMID- 2545021 TI - Malignant testicular germ cell tumor in father and son: a case report. AB - This is the eighteenth case of testicular tumor in a father and son reported in the literature. The father had a seminoma and the son an embryonal carcinoma. The trend favoring more malignant tumors occurring at younger ages in the sons is confirmed by this report. PMID- 2545023 TI - [Characterization of changes in the activity of K+-n-nitrophenylphosphatase of erythrocyte ghosts under the influence of ouabain]. AB - Variation of K+-p-NPP-ase of human ghosts under the action of ouabain (10(-11) 10(-3) M) has been studied. In human ghosts the activity of ouabain-sensitive K+ p-NPP-ase has been found to make up 65% of the total enzyme activity. The activity of ouabain-sensitive K+-p-NPP-ase reaches a maximum level at pH 7.6-8.0. A decrease in the activity of this enzyme caused by ouabain is of two-phase character. In the range of ouabain concentration from 10(-10) M to 10(-6) M and from 10(-5) M to 10(-3) M the enzyme activity lowers significantly; in the range of 10(-7) M to 10(-5) M it reaches the plato. Two types of the enzyme are assumed to exist differing by 4-5 orders of magnitude in their sensitivity to ouabain, inhibitor affinity constants and Michaelis constants. PMID- 2545024 TI - Paracrystalline inclusions in metaplastic ciliated cells of the human gastric mucosa. An ultrastructural study. AB - Unusual electron-dense paracrystalline inclusions were found in metaplastic ciliated cells in the stomachs of three Japanese male patients with gastric carcinoma. These patients had not been given antitumour drugs before surgery and ethrane (enflurane) was used as the anaesthetic. Ciliated cells in the gastric mucosa are found not infrequently in the pyloric glands in association with intestinal metaplasia in elderly Japanese patients. Paracrystalline inclusions were found only in the ciliated cells and never in any other types of gastric mucosal cell. These inclusions were located in the apical portion of the ciliated cells in intimate association with the basal bodies. They consisted of twisted strings about 27 nm wide with a regularly repeated spacing of about 30 nm. On highly magnified electron micrographs, granules about 4 nm in diameter were detected. These paracrystalline inclusions have never been reported previously, although their location in ciliated cells and their morphological characteristics suggest an intimate relationship with the ciliogenesis of metaplastic ciliated cells in the human stomach. PMID- 2545025 TI - Avian cells expressing the Newcastle disease virus hemagglutinin-neuraminidase protein are resistant to Newcastle disease virus infection. AB - The cDNA derived from the Newcastle disease virus (NDV) hemagglutinin neuraminidase (HN) gene was inserted into a replication-competent Schmidt-Ruppin Rous sarcoma virus-derived vector. Chick embryo cells transfected with this vector expressed HN-sized protein which could be precipitated with anti-HN antibody. These cells adsorbed avian red blood cells and the cell surfaces exhibited neuraminidase activity while cells transfected with an antisense version of the gene were negative for hemadsorption and neuraminidase. The cells transfected with the retroviral vector containing the HN gene were resistant to infection by NDV and influenza virus, viruses which bind to sialic acid containing receptors, but sensitive to vesicular stomatitis virus (VSV). Cells transfected with the antisense version of the HN gene were sensitive to NDV, influenza virus, and VSV infection. Thus the HN protein-expressing cells are likely resistant to NDV and influenza virus due to the destruction of the cellular receptors by the neuraminidase of the HN protein. The expression of the influenza virus HA protein using the same retrovirus vector has been reported previously (L. A. Hunt, D. W. Brown, H. L. Robinson, C. W. Naeve, and R. G. Webster, 1988, J. Virol. 62, 3014-3019). Cells infected with this vector were sensitive to infection with influenza virus, NDV, and VSV. Thus expression of a viral surface protein does not necessarily confer resistance of the cell to the homologous virus. PMID- 2545026 TI - Nucleotide sequence of bovine rotavirus gene 1 and expression of the gene product in baculovirus. AB - The nucleotide sequence of the gene that encodes for the structural viral protein VP1 of bovine rotavirus (RF strain) has been determined. The sequence data indicate that segment 1 contains 3302 bp and is A + T rich (65.3%). The positive strand of segment 1 contains a single open reading frame that extends 1088 codons and possesses 5'- and 3'-terminal untranslated regions of 18 and 20 bp, respectively. The first AUG conforms to the Kozak consensus sequence and if utilized, would yield a protein having a calculated molecular weight of 124,847, very close to the apparent molecular weight of VP1 (M.W. 125,000). The deduced amino acid sequence presents significant similarities with RNA-dependent RNA polymerase of several RNA viruses. VP1 was also synthesized in baculovirus using two transfer vecors: pAC461 and pVL941. Following infection of Sf9 cells with a recombinant baculovirus, a full-length nonfusion protein was synthesised which shares properties with authentic VP1 made in monkey kidney cells. The level of VP1 synthesis was about 10-fold higher when the baculovirus recombinant was derived from the pVL941 transfer vector. In that case, VP1 was expressed in yields approximately equivalent to 10% of the cellular protein. The recombinant protein was immunoprecipitated by hyperimmune serum raised against purified rotavirus. It also was immunogenic; a hyperimmune serum made in guinea pigs reacted with VP1 using immunoprecipitation and Western blot. This serum did not possess neutralization activity. PMID- 2545027 TI - Molecular cloning of the gene encoding the putative polymerase of mouse hepatitis coronavirus, strain A59. AB - Complementary DNA (cDNA) libraries were constructed representing the genome RNA of the coronavirus mouse hepatitis virus, strain A59 (MHV-A59). From these libraries clones were selected to form a linear map across the entire gene A, the putative viral polymerase gene. This gene is approximately 23 kb in length, considerably larger than earlier estimates. Sequence analysis of the 5' terminal region of the genome indicates the presence of the 66-nucleotide leader that is found on all mRNAs. Secondary structure analysis of the 5' terminal region suggests that transcription of leader terminates in the region of nucleotide 66. The sequence of the first 2000 nucleotides is very similar to that reported for the closely related JHM strain of MHV and potentially encodes p28, a basic protein thought to be a component of the viral polymerase (L. Soe, C. K. Shieh, S. Baker, M. F. Chang, and M. M. C. Lai, 1987, J. Virol., 61, 3968-3976). Gene A contains two of the consensus sequences found in intergenic regions. One is adjacent to the 5' leader sequence and the other is upstream from the initiation codon for translation of gene B. PMID- 2545028 TI - The open reading frame S of visna virus genome is a trans-activating gene. AB - Soon after infection of ovine cell cultures, visna virus expression is first indicated by the accumulation of two multi-spliced transcripts of 1.2 and 1.6 kb that at present we have renamed 1.4 and 1.7 kb according to their exact length. The early 1.4-kb mRNA encodes for a protein which increases the level of transcripts directed from visna virus long terminal repeat (trans-activation). This trans-activating protein was previously called VEP1 and at present is renamed as the product of the rev gene according to significant amino acid sequence homologies between this protein and the rev gene products of simian immunodeficiency virus and human immunodeficiency virus type 2. In this study, the 1.7-kb mRNA was cloned, sequenced, and in vitro translated. It is 1491 nucleotides long, contains two short open reading frames, (orfs), tat (previously orf S) and rev which is the bipartite trans-acting gene specific for the early 1.4-kb mRNA. The tat gene of visna virus encodes for a protein of 11 kDa which in transient expression assays has a positive transacting effect on transcription as the rev gene product does. PMID- 2545029 TI - Hog cholera virus--characterization of specific antiserum and identification of cDNA clones. AB - A specific antiserum was raised against the pestivirus inducing hog cholera (hog cholera virus, HCV). Using immunoprecipitation and SDS-PAGE, this antiserum served for comparison of HCV-induced proteins with those from a related and better characterized pestivirus, bovine viral diarrhea virus (BVDV). In addition to immunological relationships, the apparent molecular weights of some proteins induced by both viruses were quite similar. HCV genomic RNA was found to be about 12 kb in length, comparable to BVDV RNA. cDNA was synthesized starting from RNA isolated from partially purified virions and cloned in lambda gt11. Screening with the antiserum resulted in identification of several positive clones. Partial sequencing of one HCV-derived cDNA clone revealed a high degree of homology to a portion of the BVDV sequence. PMID- 2545030 TI - Replication of a human parvovirus nonsense mutant in mammalian cells containing an inducible amber suppressor. AB - When recombinant plasmids containing the entire adeno-associated virus (AAV) genome are transfected into permissive cells infected with a helper adenovirus, infectious AAV particles are efficiently generated. These plasmids can be used to generate mutant AAV genomes or recombinant AAV vectors. Packaging of mutant AAV genomes has required complementation with a second AAV plasmid in the transfection assay which may lead to generation of significant amounts of wild type AAV recombinants. One approach to alleviate this problem was to generate conditional lethal mutants. We constructed an AAV plasmid recombinant having a nonsense mutation in the AAV rep gene by using oligonucleotide-directed mutagenesis to convert a serine codon to an amber codon. We show that this mutant AAV can be grown on monkey cell lines containing an inducible human serine tRNA amber suppressor. The amber suppression is quite efficient and yields a burst of mutant AAV particles at about 10% of the titer of wild-type AAV. The reversion frequency of the amber mutation appears to be less than 10(-5). PMID- 2545031 TI - Two newly identified human papillomavirus types (HPV 40 and 57) isolated from mucosal lesions. AB - Two new papillomaviruses, HPV 40 and HPV 57, were isolated from a PIN lesion and an inverted papilloma of the maxillary sinus, respectively. HPV 40 showed a 13% homology to HPV 7 by reassociation kinetics and HPV 57 showed a 17% homology to HPV 2 and 25% homology to HPV 27. Hybridization of the DNA of these papillomaviruses to a wide variety of different tumor biopsies revealed that HPV 40 was present in a few genital condylomata acuminata as well as in bowenoid lesions. HPV 57 DNA was present in an oral wart, a genital condyloma acuminatum, and verrucae vulgares lesions from two immunosuppressed patients. PMID- 2545032 TI - Virus-specific polypeptides of human parainfluenza virus type 4 and their synthesis in infected cells. AB - We have studied the structural components of human parainfluenza virus type 4A (PIV-4A) and identified some virus-specific polypeptides by immunoprecipitation with polyclonal and monoclonal antibodies followed by one- or two-dimensional SDS PAGE. HN polypeptides existed as monomer, disulfide-linked dimer, and disulfide linked larger oligomer in cells infected with PIV-4A. Interestingly, the nonreduced NP, the nonreduced fusion, and the reduced F1 proteins migrated as doublets. Two F1 polypeptides were derived from different F1 + 2 proteins which migrated separately under nonreducing condition. In Vero cells infected with two strains of PIV-4A, two lower-molecular-weight proteins related to NP were detected. Oligopeptide patterns of the lower-molecular-weight protein were similar to those of NP protein synthesized in primary monkey kidney cells. The NP related low-molecular-weight protein(s) was immunoprecipitated by 1 of 11 monoclonal antibodies against mumps virus NP protein. The MAb also reacted with NP proteins of PIV-2 and SV5. Thus, the epitope recognized by the MAb was common among PIV-2, PIV-4, mumps virus, and SV5, suggesting that the epitope might have an important biological function. However, the MAb did not react with the intact NP protein from cells infected with PIV-4, indicating that the epitope of PIV-4A was presented only when NP was cleaved. Phosphorylation was demonstrated for NP and P proteins. PMID- 2545033 TI - Crystallization of Sendai virus HN protein complexed with monoclonal antibody Fab fragments. AB - The hemagglutinin-neuraminidase (HN) protein of Sendai virus has been isolated from virus particles in a biologically active soluble form after removal by proteolytic digestion of the hydrophobic amino-terminal anchor sequence (S. D. Thompson, W. G. Laver, K. G. Murti, A. Portner, J. Virol., 62, 4653-4660, 1988). The soluble HN exists as both dimers and tetramers, and crystallization trials with each of these forms have so far yielded amorphous material. Dimers complexed with Fab fragments of a monoclonal antibody formed long needle crystals. So far, these are not suitable for X-ray diffraction analysis but the results suggest that HN molecules from paramyxoviruses, even if not crystallizable, may, when complexed with Fab fragments, in some cases yield crystals suitable for X-ray diffraction analysis. PMID- 2545034 TI - Characterization of an HPV type 11 isolate propagated in human foreskin implants in nude mice. AB - Human papillomavirus type 11-Hershey (HPV-11-H) from an extract of genital warts has been serially passaged by infecting human foreskin chips that are then implanted in athymic mice (J. W. Kreider, M. K. Howett, A. E. Leure-Dupree, R. J. Zaino, and J. A. Weber (1987), J. Virol. 61, 590). Subsequent attempts to propagate HPVs present in other human lesions have not been successful. In an effort to identify the basis for the seemingly unique ability of HPV-11-H to propagate in the xenografts, we carried out extensive physical and functional characterizations of the cloned DNA, including restriction digestions, DNA sequencing of transcriptional control regions, and E2 trans-activation of the upstream regulatory region. We uncovered no significant differences compared to the prototype HPV-11 (K. Dartmann, E. Schwarz, L. Gissmann, and H. zur Hausen (1986) Virology 151, 124; H. Hirochika, T. R. Broker, and L. T. Chow (1987) J. Virol. 61, 2599). Moreover, viral enhancer assays indicated that the observed stimulatory effect of pregnancy on condylomata and of estradiol on experimental cysts is likely an indirect one and could not be attributed to up-regulation of the HPV-11 transcriptional enhancer. PMID- 2545035 TI - Identification and characterization of the BPV-2 L2 protein. AB - The bovine papilloma virus type 2 (BPV-2) L2 open reading frame was cloned into a lambda pL promoter expression vector. This plasmid was shown to express a fusion protein which constituted 75% of the BPV-2 L2 ORF linked to the first 13 N terminal amino acids of the lambda cll gene product. Antisera generated against this fusion protein were used to identify the L2 gene product as a 64,000-Da protein in BPV-2 virions. Western blot analysis demonstrated that the L2 viral protein was present in full capsids and in small amounts in empty capsids. Densitometer analysis indicated that the L2 protein constituted only 8% of the total L1 + L2 protein content of full capsids. Antisera was also used to demonstrate that the BPV-2 L2 protein is antigenically related to the BPV-1 L2 protein. PMID- 2545036 TI - Nucleotide sequence of human papillomavirus type 31: a cervical neoplasia associated virus. AB - The nucleotide sequence of human papillomavirus (HPV) 31 DNA (7912 bp) was determined and used to deduce the genomic organization of this cervical cancer associated virus. Based on comparisons of the HPV 31 DNA sequence to other sequenced HPVs, HPV 31 is a typical papillomavirus most related to HPV 16 (70% identical nucleotides). The E6 and E7 open reading frames (ORF) of HPV 31 contain several potential DNA binding motifs (Cys-X-X-Cys), the locations of which are conserved in all HPVs. The E6 ORF also has the potential to code for an E6* protein. The E7 ORF of HPV31 encodes a polypeptide motif which appears to distinguish HPVs associated with cervical cancer, such as types 16, 18, 31, and 33, from HPVs found primarily in benign lesions, such as types 6 and 11. PMID- 2545037 TI - Physical mapping of the human cytomegalovirus (HCMV) (Towne) DNA polymerase gene: DNA-mediated transfer of a genetic marker for an HCMV gene. AB - DNA-mediated transfer of a drug resistance marker (phosphonoacetate resistance) for the HCMV (Towne) DNA polymerase (pol) gene has been used to genetically confirm the physical localization of the HCMV (Towne) DNA pol gene. The HCMV (Towne) genomic region homologous to the pol genes of other herpesviruses was first identified by moderate stringency Southern hybridization. Restriction fragments from this region were molecularly cloned from previously characterized phosphonoacetic acid resistant (PAAr) HCMV genomes (R. T. D'Aquilla and W. C. Summers, J. Virol., 61, 1291-1295, 1987). A high frequency of recombinant PAAr virus was found among the progeny of cotransfections of infectious, wild-type HCMV (Towne) DNA only with pol-homologous restriction fragments cloned from PAAr HCMV. The co-transfection technique described here may facilitate further gene mapping in HCMV. The results presented here provide functional proof that the HCMV pol gene is encoded by the sequences previously identified as homologous to other herpesvirus pool genes. PMID- 2545038 TI - Extensive antigenic diversity among human parainfluenza type 2 virus isolates and immunological relationships among paramyxoviruses revealed by monoclonal antibodies. AB - A panel of 128 monoclonal antibodies (MAbs) directed against hemagglutinin neuraminidase (HN), fusion (F), matrix (M), and polymerase (P) proteins, and nucleoprotein (NP) of the Toshiba strain of human parainfluenza type 2 virus (PIV2) was prepared to examine the antigenic relationships among clinical isolates of PIV2 and among paramyxoviruses by indirect enzyme-linked immunosorbent assays. The HN proteins of 18 clinical isolates of PIV2 showed extensive antigenic diversity: 23 of 33 anti-HN MAbs showed no or limited reactivity to many isolates, while other structural proteins were antigenically well conserved. Some anti-HN MAbs recognizing conserved epitopes of the isolates exhibited two types of neutralizing activity, that is, these antibodies inhibited viral infectivity through attachment inhibition or fusion inhibition. This result also showed the presence of a potential third function of the HN protein which might affect the fusing activity of the F protein besides the hemagglutinating and neuraminidase activities. Many of the anti-NP and anti-P MAbs reacted with simian virus 41 (SV41) and simian virus 5 (SV5), whereas a few reacted with mumps virus or PIV4. Two of 6 anti-F MAbs reacted with SV41. None of the 128 MAbs showed reactivity with PIV1, PIV3, Newcastle disease virus (NDV), and measles virus. This result confirmed antigenic proximity of SV5 and SV41 to PIV2 and revealed comparatively restricted immunological relatedness among PIV2, PIV4, and mumps virus. PMID- 2545039 TI - Eclipse products of poliovirus after cold-synchronized infection of HeLa cells. AB - The particles derived from radioactively labeled poliovirus, in cold-synchronized infection of HeLa cells, were studied. After temperatures shift-up, radioactivity was transferred rapidly from the infecting virions (160 S) to 135 S, and from there more slowly to 110 S, a previously unreported eclipse product. Both the 135 and 110 S particles contained RNA, lacked VP4, and were H-antigenic. In the 135 S particles, part of the VP1 complement was lost; in addition to this, the VP2 polypeptide of the 110 S particles was cleaved in situ. The formation of eclipse products was insensitive to cycloheximide. Arildone totally blocked the formation of 135 S particles, but not their further processing. PMID- 2545040 TI - Interaction of rotavirus cores with the nonstructural glycoprotein NS28. AB - The nonstructural rotavirus receptor glycoprotein NS28 is 175 amino acids long and oriented in the RER membrane with the NH2 terminus on the luminal side and approximately 131 amino acids accessible from the cytoplasmic side. Au et al. (1988) have demonstrated that NS28 is able to interact with rotavirus single shelled particles (cores) in a receptor:ligand interaction in which NS28 appears to act as the receptor and the rotavirus core as the ligand. This interaction appears to model the events that occur in the infected cell in which virus maturation involves budding of the core into the lumen of the RER. We have investigated the nature of the interaction between cores and NS28 in vitro using membranes derived from SA11 rotavirus-infected MA104 cells and membranes from cells where NS28 and other rotavirus proteins have been expressed using a series of recombinant vaccinia viruses that incorporate appropriate cloned rotavirus genes. The interaction between the core and the receptor is enhanced by the presence of Ca2+ and Mg2+ and Scatchard analysis yields a dissociation constant (Kd) of 5 x 10(-11) M. The major core protein VP6 is the ligand involved because (i) a monoclonal antibody specific for VP6 blocks the reaction, (ii) membranes prepared from cells infected with a double recombinant vaccinia virus which expresses both NS28 and VP6 exhibit a reduced capacity to bind cores, and (iii) VP6 prepared from virus blocks the ability of membranes to bind cores. When VP6, VP7, VP4, and NS28 are expressed singly as the sole viral proteins present in the cell, only membranes from cells expressing NS28 mediate receptor function, indicating that the presence of NS28 is sufficient to mediate the interaction between cores and the membrane and that other viral proteins probably are not involved in the initial receptor:ligand interaction. PMID- 2545041 TI - [Metabolism and reception of vitamin D]. PMID- 2545043 TI - [Status of the antioxidant system of erythrocytes in newborn infants in acute and chronic hypoxia]. AB - Low activity of catalase, superoxide dismutase, alterations in metabolism of glutathione as well as activation of lipid peroxidation and increase in Na+, K+ ATPase activity were found in erythrocytes of newborns funic blood under conditions of chronic hypoxia and simultaneous effects of acute and chronic hypoxia. Acute hypoxia during labor caused increase in activity of superoxide dismutase, catalase, glucose-6-phosphate dehydrogenase, Na+, K+-ATPase as well as in the rate of lipid peroxidation. These alterations, observed in the enzymatic activity and in the rate of lipid peroxidation, which were detected in funic blood erythrocytes under conditions of hypoxia, may be responsible for some diseases during postnatal period of children ontogenesis. PMID- 2545042 TI - [Adrenergic regulation of the level of enkephalins in the myocardium]. AB - Content of enkephalins was increased in myocardium after administration of tyramine (liberator of endogenous catecholamines). Neoepinephrine decreased content of Met-enkephalin in heart. Catecholamines circulating in blood exhibited less distinct effect on content of opioid peptides in myocardium as compared with endogenous free catecholamines. Inverse correlation was found between content of cAMP and Met-enkephalin in myocardium. All the pharmacological drugs studied could exhibit the mediatory effects on opioid peptides in heart via alterations in activity of vegetative nervous system affecting the myocardium. PMID- 2545044 TI - [The effect of eicosapentaenoic acid and various prostaglandins on cholesterol accumulation and proliferation of intima cells from human aorta]. AB - Consumption of 15 g of 40% eicosapentaenic acid within 3 days resulted in 3-fold increase of its content in blood serum of healthy persons and of patients with heart ischemic disease, whereas concentration of arachidonic acid was unaltered. Administration of blood serum containing increased content of eicosapentaenic acid into cell culture of human aorta intima caused a decrease in accumulation of the intracellular cholesterol and in 3H-thymidine incorporation. Simultaneous addition of the blood serum containing eicosapentaenic acid and the structure derivative of A2 thromboxane V 46619 into the aorta intima cell culture led to a distinct decrease in the atherosclerotic effect of the latter drug. Combination of the blood serum with carbocycline or with prostaglandin I2 resulted in the most pronounced lowering of intracellular cholesterol content and of the cell proliferating activity. The antiatherosclerotic effect of eicosapentaenic acid appears to relate not only to the fact that the drug is precursor of prostaglandine I2 and I3 but also to the regulating effect on a balance of various prostaglandins. PMID- 2545045 TI - [The enzymatic activity of DNA metabolism of the blood in patients operated on in stomach cancer]. AB - The study was concerned with comparison of the activity of thymidine kinase, thymidine phosphorylase, adenosine desaminase and AMP 5'-nucleotidase in serum of cancer patients and healthy subjects. A significant change in 5'-nucleotidase level was registered in patients over 60 years of age and in those of adenosine desaminase and thymidine phosphorylase in patients older than 70 years. A comparative evaluation of the activity of those enzymes showed an increase in thymidine kinase activity matched by a sharp drop in that of thymidine phosphorylase in patients over 45 years. A sharp increase in the activity of adenosine metabolizing enzymes in cancer serum was accompanied by its decrease in lymphocytes. Among surgical patients, significant changes in the activity were observed in radically operated cases only. PMID- 2545046 TI - [Effect of progestin therapy on the level of cytoplasmic receptors in patients with cancer of the corpus uteri]. AB - Correlations were identified between tumor morphobiochemical characteristics and basic parameters of reproductive homeostasis in 256 cases of endometrial carcinoma. A correlation was established between the levels of cytoplasmic receptors to estradiol (ER) and progesterone (PR), on the one hand, and morphological differentiation and spontaneous secretion in tumor tissue, on the other. A direct correlation was found between ER and metastatic spread to regional lymph nodes, and no correlation--between ER and PR and major parameters of reproductive homeostasis. A significant decrease in ER, PR, gonadotropins, estradiol and colpocytological reaction indexes followed hormone therapy with hydroxyprogesterone capronate and medroxyprogesterone acetate. The criteria of endometrial carcinoma patients sensitivity to progestin therapy were as follows: ER and PR levels--more than 30.0 fmol/mg protein, estradiol--more than 30-50 pg/ml and colpocytological index--over 40%. PMID- 2545047 TI - [Characteristics of the postoperative period in stomach cancer patients undergoing combined treatment]. AB - Postoperative course was studied in 423 radically treated patients: 175--surgery only, 96--preoperative 5-fluorouracil chemotherapy, 79--intravenous radioactive colloid gold postoperatively, and 73--both 5-fluorouracil preoperatively and colloid gold postoperatively. The two latter procedures did not affect postoperative course adversely; nor did they influence the spectrum of postoperative complications or increase lethality rate. PMID- 2545048 TI - Chronic fatigue syndrome. A critical appraisal of the role of Epstein-Barr virus. AB - The symptom complex currently designated the chronic fatigue syndrome was previously termed the chronic or chronic active Epstein-Barr virus syndrome or the chronic mononucleosis syndrome, prematurely assuming an etiologic role for the Epstein-Barr virus (EBV). This presumption derived from the fact that some patients with the chronic fatigue syndrome have very high or very low titers of certain antibodies to EBV. A review of seroepidemiologic patterns of response to EBV and of studies of patients with the chronic fatigue syndrome shows that these antibody titers overlap considerably both with those of controls or other healthy persons and with those of patients with other illnesses. Given the high prevalence of exposure to EBV, it would be difficult to determine whether the virus caused the syndrome or whether the antibody elevations resulted from the illness, even if distinct differences in titers existed. Other methodologic issues of control selection, laboratory test comparability, and differing case definitions pose problems in studying this syndrome. The recently published working case definition should facilitate the continuing search for causes. PMID- 2545049 TI - [Stereotaxic biopsy of intracranial processes: validity of histologic diagnosis]. AB - The diagnostic validity of stereotactic intracranial biopsies was investigated in 70 patients retrospectively. In 56 cases (80%) the presence of a neoplastic lesion as well as its grade of malignancy was proved by cytological, histological and immunohistochemical techniques. An inflammatory or vascular lesion was found in 8 patients (11.4%). In 6 patients (8.6%) the nature of the lesion remained unclear because of nonspecific histological findings. In 27 cases a correlation was found between the biopsy specimens and corresponding material obtained during open surgery or autopsy. 24 concordant results were found (88.9%). In one case a malignant tumour was classified only according to the findings obtained at operation. In two cases subsequently diagnosed as glioblastoma biopsy reported a higher differentiated astrocytoma and a non-neoplastic lesion, respectively. These results confirm stereotactic biopsy of intracranial lesions as a method with low complication rate and high diagnostic validity. PMID- 2545050 TI - [Regulation of intracellular pH by Na+/H+ exchange in human lymphocytes]. AB - Spectrofluorimetry and the pH-sensitive fluorescent dye 2',7'-bis(carboxyethyl) 5(6)carboxyfluorescein (BCECF) was used to measure the intracellular pH (pHi) of suspended human lymphocytes. A linear relationship exists between pHi and the fluorescent spinal-ratio (I490nm/I435nm, emission 526 nm) between pH 6.5 and pH 7.8. At the end of each experiment the ratio was calibrated using the high [K+] nigericin technique. All solutions were HEPES buffered. The pHi in resting cells was 7.27 +/- 0.02 (n = 37) at 25 degrees C. Na+ free solution caused a pHi decrease to 6.81 +/- 0.08. The ammonium prepulse technique (25 mM NH4Cl) dropped the pHi to pH 6.80. A rapid recovery of the pHi after this acidification was observed in NaCl Ringer solution. Na+ free solution completely blocked the recovery. 1 mM amiloride led to a partial block of recovery. pHi was restored to the basal value after readdition of Na+. We conclude that in HEPES buffered solutions human lymphocytes recover pHi via a mechanism dependent on extracellular Na+ and largely accomplished by an amiloride inhibitory Na+/H+ exchanger. PMID- 2545052 TI - Adult Wilms' tumor--a case report. AB - A case of Wilms' tumor in a 67 year-old female is presented. The tumor totally replaced the left kidney with extension to Gerota's fascia, the adrenal gland and ureter, with tumor thrombi in the left renal vein. Multiple bony metastases to the skull, shoulder, rib, ilium, acetabulum and femur were noted also. Microscopically, the tumor was composed predominantly of blastemal tissue with diffuse anaplasia with areas of tubular differentiation and chondroid elements. Clear cell nests were found in the proximal ureter with gradual maturation downward. PMID- 2545051 TI - Phase II study of cyclophosphamide, doxorubicin, and vincristine (CAV) and etoposide plus cisplatin (EP) alternating chemotherapy combined with radiotherapy in small cell lung cancer. AB - The development of drug resistance is the major limiting factor influencing the survival of patients with small cell lung cancer (SCLC). We have thus examined the activity of cyclophosphamide, doxorubicin and vincristine (CAV) alternating with etoposide and cisplatin (EP) in 35 patients with SCLC. The treatment courses were alternated every 3 or 4 weeks. After induction chemotherapy, patients with limited disease (LD) received thoracic radiotherapy (5000 cGy), prophylactic cranial irradiation (3000 cGy) and maintenance chemotherapy and patients with extensive disease (ED) received maintenance chemotherapy only. In this group of 35 patients, 13 had limited disease (LD) and 22 had extensive disease (ED). After completion of the therapy, 100% of the patients with LD achieved complete plus partial remission (CR + PR) and 68% of the patients with ED achieved CR + PR. The median survival time was 66 weeks (15.3 months) in patients with LD and 44 weeks (10.2 months) in patients with ED. The over all survival for patients with LD was superior to that for patients with ED (p less than 0.05). Also, median response duration for patients with LD (35 wks) was longer than that for patients with ED (17 weeks) (p less than 0.05). The primary site was the most vulnerable site to relapse (18 patients). Toxicity was mild to moderate and acceptable, and there were no treatment-related deaths. These results suggest that the alternation of CAV and EP is effective treatment strategy in the management of SCLC. A randomized controlled study will be required to discriminate the actual effect of this alternating regimen. PMID- 2545053 TI - [Focal nodular hyperplasia of the liver]. AB - Among the benign liver tumours particularly the focal nodular hyperplasia (FNH) increased its importance. Within 10 years we could observe 28 patients with the lesion. Macroscopically the nodular changes show extremely characteristic findings. The histological picture reminds us of cirrhosis, the prevailing presence in women and the clinical recognition mostly at reproductive age let us think of a hormonal action in development and growth of the lesion. The same concerns the use of hormonal contraceptive drugs. Differential-diagnostically particularly malignant processes are to be demarcated. Though the danger of a haemorrhage with formation of a haemoperitoneum is insignificant, in the individual case a surgical approach should be taken into consideration. PMID- 2545054 TI - Prevention of postoperative deep vein thrombosis by one daily injection of low molecular weight heparin and dihydroergotamine. AB - 201 patients aged over 40 years undergoing abdominal surgery were divided randomly into two groups. Each patient received two subcutaneous injections daily: the first group received a morning injection of 1500 aPTT U of low molecular weight heparin combined with 0.5 mg dihydroergotamine (LMH/DHE) and an evening injection of placebo; the second group received morning and evening injections of 2500 IU standard heparin combined with 0.5 mg dihydroergotamine(H/DHE). 25 patients were withdrawn during the course of the trial, 13 in the LMH/DHE group and 12 in the H/DHE group. There was no significant difference between the two groups with regard to age, sex, body weight or history of thromboembolism. 125I-labelled fibrinogen test was routinely used to detect deep vein thrombosis (DVT), which was confirmed by phlebography. Ventilation-perfusion scanning was performed in patients in whom pulmonary embolism was suspected on clinical grounds. DVT occurred in 6 patients in the LMH/DHE group (6.9%) and in 7 patients in the H/DHE group (7.9%). Pulmonary embolism occurred in one patient in each group. The only noteworthy haemorrhagic incident was a haematoma of the abdominal wall in one patient (LMH/DHE). It was concluded that a single daily injection of 1500 aPTT U low molecular weight heparin combined with DHE is as effective and as well tolerated as two injections daily of 2500 IU standard heparin combined with DHE. PMID- 2545055 TI - [Aneurysmic transformation of the venous system in venous angiodysplasias of the limbs]. AB - Based on a retrospective evaluation of 107 patients with congenital venous angiodysplasia of the Type Klippel-Trenaunay (n = 76) and Type Servelle-Martorell (n = 31) the frequency and pathogenesis of aneurysm formation in the venous system has been analysed. The vascular patterns include both cylindric ectasias and fusiform aneurysms with an incidence of approximately 40%. Preferred locations are subcutaneous drainage veins, the popliteal, external iliac vein and atypic communicating veins between the superficial and the deep venous system. Complications of the aneurysm such as local thrombosis, recurrent pulmonary embolism or bleeding from rupture were not observed. From a pathogenetic point of view the aneurysm formation in venous angiodysplasias results probably from two causative factors, i.e., a congenital weakness of the venous wall (inborn error?) and an abnormal hemodynamical stress situation. The latter is caused by concomitant malformations of the deep venous system (avalvulia, hypo- and/or aplasia). The persistent intermittent venous hypertension associated with a more or less pronounced increase of the venous volume in the affected venous system of the limb results in a deep venous insufficiency respectively venous reflux disease. Surgery is indicated under two conditions: a) in the presence of aneurysm complications or b) for the elimination of a pathological short circuit flow in some drainage veins. Antireflux surgery, e.g., venous valve transfer form the brachial vein, is up to recently still in a stage of experimental-clinical investigation. The therapy of choice is predominantly conservative, i.e., external compression bandages or stockings to reduce the deleterious effects of a chronic deep venous insufficiency respectively venous reflux disease. PMID- 2545056 TI - [Experimental embryologic, biochemical and molecular biology approaches to the identification of embryonic inducers]. AB - Problems of the mechanisms of embryonic induction in vertebrate development have been considered on the basis of author's experimental data. Though several polypeptide factors with certain inducing activity have been identified recently, molecular genetic mechanisms of their effect on embryonic target cells remains largely unclear. One of possible causes of very slow progress in this area of developmental biology is an inadequate system of biotesting of inducers at tissue level (ectoderm of early amphibian gastrulae) using histological criteria. A necessity for carrying out similar studies on cellular level and estimating effect of inducers using immunochemical and molecular biological methods has been postulated. Methods allowing to carry out biotesting of inducers on cell suspension or aggregate of a one type of embryonic cells have been proposed. New approaches, combining the methods of experimental embryology and molecular biology, to studies of embryonic inducers, receptors, and their mRNA, have been analyzed. PMID- 2545057 TI - [Possible involvement of the renin-angiotensin system in reproduction. II. Occurrence and role in the female reproductive tract]. AB - Biochemical and immunological studies of the last years reveal the existence of an "ovarian renin-angiotensin system (RAS)". Despite of the low angiotensin conterting enzyme (ACE) activity in the ovary the follicular fluid is rich in angiotensin II (AII). The detection of AII receptors on cells within maturating follicles proves them as AII targets. Therefore, it is supposed that AII may be involved in the regulation of fundamental processes of follicle maturation and/or corpus luteum formation. Further interesting findings are the high concentration of prorenin in the follicle fluid of women causing an increase of the prorenin blood plasma level at time of ovulation, and a second increase of the blood prorenin concentration in the middle of the luteal phase. With respect to the ACE activity in the ejaculate it is imaginable that the smooth muscle tonus of the uterus and the oviduct could be affected by local generation von AII and/or degradation of bradykinin and thus the transit of the semen may be facilitated. Further systematic research is necessary to bring more light into the physiological context and to replace hypothetical interpretations of the findings by exact knowledge. PMID- 2545058 TI - [Design of a nutrient medium for bacteria of the genus Haemophilus--producers of restriction endonucleases]. AB - A culture medium for the cultivation of hemophilic bacteria, containing acidic casein hydrolysate, aminopeptide and fodder yeast extract, has been proposed. The growth-stimulating properties of this medium have been studied on 5 strains producing restrictases differing in their specificity. In growing these producer strains in a model ANKUM-2 fermenter with the supply of carbohydrate substrates (glucose, sucrose, glycerin) the yield of biomass, considered to be high for hemophilic bacteria (10-14 g of humid substance from 1 liter of the medium), has been achieved. As shown on H. influenzae Rc B-2297 used as an example, an increase in the yield of microbial biomass leads to a decrease in restrictase specific activity. PMID- 2545059 TI - [Human infectivity with enterobacteria and enteroviruses while swimming depending on the level of microbial water pollution]. AB - The microbiological study of the microflora of the intestinal contents taken from children and water used for bathing has revealed the dependence of the risk of infection of humans with the causative agents of acute intestinal diseases on the level of the microbial contamination of water. PMID- 2545060 TI - [Immunobiologic activity of Bordetella pertussis strains defective in various virulence factors]. AB - The immunobiological properties of mutant strains, selectively deprived of certain antigens (hemagglutinin, B. pertussis toxin, dermonecrotic toxin, hemolysin, adenylate cyclase), have been studied with the aim of finding out the relationship between the presence of certain antigens in microbial strains and their protective properties. The results of these studies suggest that the protective potency of pertussis vaccine may be related to the presence of some antigenic substances, including those not pertaining to the known factors of virulence. PMID- 2545061 TI - Expression of insulin-like growth factor receptors in primary human thyroid neoplasms. AB - The presence of IGF-I receptors was demonstrated in normal and neoplastic tissues of human thyroid. Binding of (125I)IGF-I to thyroid membranes was dependent on time and temperature of incubation, and maximal binding was achieved at 4 degree C and 18 h of incubation. (125I)IGI-I binding was dose-dependently displaced by unlabelled IGF-I; half-maximal inhibition occurred at concentrations of 10-20 milligrams. IGF-II and insulin had relative potencies of 5 and 1% compared with IGF-I. Scatchard analysis of binding data revealed a single class of IGF-I receptors with high affinity (Ka: 1.2-8.6 x 10(9) 1/mol) in normal thyroid tissues. Affinity cross-linking and autoradiography demonstrated the type IIGF receptors. Specific binding of (125I)IGF-1 in thyroid cancer tissues (9.69 +/- 2.07% per 200 micrograms protein; mean +/- SEM, N = 8) was significantly (p less than 0.05) higher than that in the surrounding normal tissues (3.03 +/- 0.35%, N = 8). In contrast there was no difference in the binding between adenoma tissues (4.19 +/- 0.53%, N = 5) and the adjacent normal tissues (2.94 +/- 0.24%, N = 5). The higher IGF-I binding in cancer tissues was due to an increase in the binding capacity without any change in the affinity. The presence of IGF-I receptors suggests a possible role of IGF-I and its receptors in the growth of thyroid cancer cells. PMID- 2545062 TI - Aggravation of hypoglycemia in insulinoma patients by the long-acting somatostatin analogue octreotide (Sandostatin). AB - Recently somatostatin analogues were successfully used to control insulin-induced hypoglycemia in patients with insulinoma. We observed a transient decrease in glucose levels and symptomatic hypoglycemia after administration of the long acting somatostatin-analogue octreotide (Sandostatin) in two insulinoma patients. We studied the acute effects of octreotide (administered before breakfast) on blood glucose and gluco-regulatory hormones in these patients. In one patient, we studied the effects of glucagon replacement and changing the time of breakfast (relative to octreotide administration) on octreotide-associated changes in blood glucose and glucoregulatory hormones. Compared with control levels, octreotide therapy reduced insulin levels. During hypoglycemia glucagon and growth hormone levels were suppressed, but cortisol levels appropriately increased. The increase in catecholamine levels was normal in one patient, but markedly attenuated in the other. A transient decrease in serum glucose after octreotide was absent after glucagon replacement, but present when breakfast was taken before administration of octreotide. We conclude that in patients with insulinoma, octreotide therapy may be associated with clinically important hypoglycemia, during which counterregulatory hormone secretion may be attenuated. PMID- 2545063 TI - Human serum LH inhibitor(s): behaviour and contribution to in vitro bioassay of LH using dispersed mouse Leydig cells. AB - The present study aimed at investigating the nature and causes of non-parallelism in testosterone responses to serial dilutions of peripheral serum and standard LH preparations in the mouse Leydig cell in vitro bioassay of LH. Immunoadsorption with monoclonal antibody to the beta-subunit of LH was used to obtain LH-free serum; the procedure removed more than 98% of the immunoassayable LH. When a constant amount of the LH-free serum was added to standard dilutions, the bioassay dose-response curves to serum dilutions, the standards became parallel, i.e. the well-known source of error of this assay system was eliminated. When standard curves prepared in medium and LH-free serum (final concentration 10%) were compared, no effect of the serum was found on basal cAMP and testosterone production. However, the LH-stimulated testosterone and cAMP production were suppressed by serum by a rather constant factor of 40%. Mild heating (60 degrees C, 15 min) or treatment with dextran-coated charcoal, but not other extraction, was able to eliminate the inhibitory activity of the LH-free serum. Binding studies demonstrated that [125I]HCG interaction with mouse Leydig cell homogenates was inhibited by LH-free serum in a fashion indicative of reduced LH receptor number, but not of reduced binding affinity. In conclusion, these data show that human serum contains LH inhibitor(s) which affect the LH-receptor interaction and LH stimulated testosterone production in mouse Leydig cell in vitro. The effect is marked in serum concentration over 1.5% and it shows only minor variation between individual sera. This source of error can be effectively removed from the LH in vitro bioassay by using LH-free serum for preparation of dilutions of LH standards. PMID- 2545064 TI - Endocrine abnormalities in patients with adrenal tumours incidentally discovered on computed tomography. AB - To determine endocrine activity of adrenal tumours incidentally discovered on C.T., we examined 20 consecutive patients. They underwent thorough hormonal assessment and scintigraphic scanning with radioactive cholesterol under dexamethasone suppression (19 patients). Biochemical findings compatible with cortisol hypersecretion were detected in 5 patients. One patient had reduced reserves of cortisol secretion and one had hyporeninemic hypoaldosteronism. The scintigraphy showed no uptake in 10, unilateral uptake in 4, and bilateral uptake in 5 patients. In 3 patients the finding was unilateral on CT. but bilateral on scintigraphy. Signs of autonomous cortisol production were more common among patients who had uptake on scintigraphy. At the operation of 8 patients only benign lesions were found. During the follow-up (9 to 49 months) of the 12 unoperated patients, the tumour disappeared in one and remained unchanged in the others. No changes occurred in the biochemical findings except in one patient whose cortisol response to 1 mg of dexamethasone became abnormal. Since slight hypercortisolism or a bilateral disease often exists behind an incidentally discovered adrenal tumour, we emphasize the importance of careful assessment of cortisol metabolism and a scintigraphic scanning under dexamethasone suppression in the examination of these patients. PMID- 2545065 TI - The effect of thyroxine, 3, 5-dimethyl-3'isopropyl-L-thyronine and iodized oil on fetal brain development in the iodine-deficient sheep. AB - Studies have been carried out to investigate the role of maternal and fetal thyroid function in the effects of iodine deficiency on fetal brain development in sheep. Iodine deficiency was established with an especially prepared low iodine diet of maize and pea pollard. The iodine-deficient sheep were mated and the end of the second trimester of pregnancy (100 days gestation) were divided into groups which received either a sc injection of T4 or 3,5-dimethyl-3 isopropyl-L-thyromine or an injection of iodized oil. AT 140 days gestation (10 days prior to parturition) comparison of the fetuses delivered by hysterotomy revealed that the retarded fetal brain development observed in iodine deficiency was greatly improved by T4 and by iodized oil. However, T4 and iodized oil failed to correct the reduction in the number and the increase in the length of synaptic appositions which were observed in the fetal cerebral cortex after iodine deficiency. In addition, the histological appearance of the fetal thyroid gland and the levels of plasma thyroid hormones were restored to normal. The administration of 3,5-dimethyl-3'-isopropyl-L-thyronine had no effect on the retarded fetal brain and body development of the iodine-deficient fetuses. The lack of response may be due to the ability of 3,5-dimethyl-3'-isopropyl-L thyronine to cross the ovine placenta as no reduction in the abnormally elevated fetal plasma TSH observed in spite of a fall in maternal plasma TSH and apparent restoration of maternal thyroid function.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545066 TI - Plasma 18-hydroxycorticosterone and aldosterone responses to angiotensin II and corticotropin in diabetic patients with hyporeninemic and normoreninemic hypoaldosteronism. AB - To examine the nature of adrenal abnormalities in diabetic patients with hyporeninemic and normoreninemic hypoaldosteronism, responses of plasma 18 hydroxycorticosterone and plasma aldosterone to angiotension II infusions and ACTH injection were investigated in 8 diabetic patients with hyporeninemic hypoaldosteronism and 9 diabetic patients with normoreninemic hypoaldosteronism compared to 11 control subjects. In both the patients with hyporeninemic and normoreninemic hypoaldosteronism, plasma 18-hydroxycorticosterone and plasma aldosterone were low, whereas plasma cortisol and plasma corticosterone were within normal ranges. Percent increments of plasma 18-hydroxycorticosterone and plasma aldosterone above their baseline levels after angiotensin II infusions were low or somewhat low in the patients with hyporeninemic hypoaldosteronism and low in the patients with normoreninemic hypoaldosteronism. Percent increments of plasma 18-hydroxycorticosterone and plasma aldosterone above their baseline levels after ACTH injection were similar in three groups. These results suggest that in diabetic patients with isolated hypoaldosteronism, the adrenal abnormality, regardless of whether it is primary or secondary, is mainly due to impaired adrenal responsiveness to angiotension II and atrophy and the zona glomerulosa. PMID- 2545067 TI - Tunicamycin modulates binding of 125I-erythropoietin to Friend erythroleukemia cells. AB - The effects of tunicamycin and neuraminidase treatment on the specific binding of 125I-erythropoietin to a murine erythroleukemia cell clone, B8, were investigated. Neuraminidase treatment of B8 cells did not affect the specific binding of erythropoietin, but tunicamycin treatment caused a 2.5 to 4-fold increase in the amount of 125I-erythropoietin binding. Scatchard analysis of the binding data showed that the increase in the amount of binding resulted from increased affinity of the receptor. These results suggest that N-linked sugars of the erythropoietin receptor protein are involved in the interaction of erythropoietin with the cell-surface receptors on B8 cells. PMID- 2545068 TI - Endothelium and thrombomodulin: expression and distribution of thrombomodulin on altered endothelium and neoplastic syncytiotrophoblast. AB - We investigated the expression of thrombomodulin (TM) on endothelium in some pathologic states. We used the cultured endothelial cells treated with interleukin-1 (IL-1) and propagated cells by serial subculture for extended periods of time and assessed cell-surface TM molecules. We also studied the distribution of TM on surgical specimens of chorionic diseases, angiosarcoma and on several established cell lines of human choricarcinoma. Subculture of human umbilical vein endothelial cells (HUVE) up to 2 months (approximately 16 subcultures) decreased the number of cell-surface TM molecules by approximately 20% compared to the primary culture. The number of TM molecules also decreased on HUVE treated by IL-1. The treatment of the cells with IL-1 also induced change of shape. TM was found not only normal syncytiotrophoblast but also on neoplastic syncytiotrophoblast of choriocarcinoma and hydatidiform mole. However, TM was not expressed on the three established cell lines. TM was found on various types of vascular tumors, including angiosarcoma. PMID- 2545069 TI - Relationship between immunophenotype and the development of second primary neoplasms in patients with non-Hodgkin's lymphoma. AB - We reviewed the records of 107 patients with non-Hodgkin's lymphoma (NHL) to evaluate the relation between second primary neoplasms and the NHL immunophenotype. The incidence of second primary neoplasms was 3.7%. There were one case of hepatocellular carcinoma and 3 cases of gastric adenocarcinoma including one patient who had a history of metachronous malignant lymphomas. Three patients had B cell lymphoma with monoclonal IgM kappa phenotype, and one patient had follicular mixed cell type lymphoma with serum monoclonal IgM kappa. The dominant immunophenotype of B cell lymphomas in Japanese patients is IgM lambda. We believe that the association of the uncommon phenotype of IgM kappa with second primary neoplasms, especially gastric cancer, reflects an underlying genetic predisposition. NHL patients with IgM kappa phenotype should be evaluated carefully for second primary neoplasms. PMID- 2545070 TI - Histochemical detection of alpha-D-galactosidase with 5-Br-4-Cl-3-indoxyl alpha-D galactoside. AB - 5-Br-4-Cl-3-indoxyl alpha-D-galactoside was used as a new substrate in azoindoxyl, indigogenic, and tetrazolium procedures for the detection of alpha-D galactosidase in the light microscope. Compared with the simultaneous azo-dye methods using 1-naphthyl or 6-Br-2-naphthyl alpha-D-galactoside as substrates and hexazotized pararosaniline for simultaneous coupling, primarily the azoindoxyl method with hexazotized pararosaniline and the indigogenic technique proved to be superior. The azoindoxyl reaction is recommended for the localization of alpha-D galactosidase in lysosomes when freeze-dried celloidin-coated cryostat sections are used: the indigogenic procedure should be employed for the detection of the total activity of the enzyme in combination with the technique of semipermeable membranes. The tetrazolium reaction delivered high amounts of formazan as the final reaction product; however, its localization was less precise than with the azoindoxyl and indigogenic methods. Conspicious species differences were found; except for the small intestine of suckling mice the highest activities of alpha-D galactosidase were present in different immature and mature rat organs when compared with mouse, hamster, guinea-pig, marmoset monkey, and human tissues. PMID- 2545071 TI - Zinc, copper and magnesium concentration in serum and CSF of patients with neurological disorders. AB - Zinc (Zn), copper (Cu) and magnesium (Mg) concentrations in cerebrospinal fluid (CSF) and serum were determined with atomic absorption spectrophotometry in 74 patients suffering from various neurological diseases, and in 28 healthy controls. Increased CSF zinc levels were found in the group of peripheral nervous system diseases (P less than 0.01) and in the cases of different neurological syndromes with increased CSF protein concentration (P less than 0.001). Increased CSF and serum copper levels were found in the cases with increased CSF protein levels (P less than 0.05). It is probable that the damaged blood-brain-barrier (BBB) permits the passage of the trace elements Zn, Cu and of Mg into the subarachnoid space. Decreased serum Cu levels (P less than 0.01) were found in the group of multiple sclerosis (MS). The findings are correlated to those of previous communications. PMID- 2545072 TI - Drug binding properties of neonatal albumin. AB - Neonatal and adult albumin was isolated by gel chromatography on Sephacryl S-300, from adult and umbilical cord serum, respectively. Binding of monoacetyl-diamino diphenyl sulfone, warfarin, sulfamethizole, and diazepam was studied by means of equilibrium dialysis and the binding data were analyzed by the method of several acceptable fitted curves. It was found that the binding affinity to neonatal albumin is less than to adult albumin for monoacetyl-diamino-diphenyl sulfone and warfarin. Sulfamethizole binding to the neonatal protein is similarly reduced when more than one molecule of the drug is bound per albumin molecule, and binding of the first sulfamethizole molecule is possibly reduced as well. Diazepam binds with equal affinity to the fetal and adult proteins. Among the two main albumin drug-binding functions, for warfarin and diazepam, the former is thus compromised in the newborn infant while the diazepam binding function is at the adult level. PMID- 2545073 TI - Endogenous digoxin-like factor in neonates: effect of age and relation to serum bilirubin levels. AB - Endogenous digoxin-like immunoreactive factor(s) (DLIF) have been found in serum and urine of newborn infants, including those born prematurely. We assessed the effect of age on serum levels of DLIF in 73 samples obtained from 66 healthy full term newborn infants at birth and during the first two months of life. DLIF concentrations were highest at birth and fell progressively with age. In cord blood, DLIF levels were 0.73 +/- 0.35 ng/ml (mean +/- SD). DLIF concentrations were 0.45 +/- 0.11 ng/ml on day 1, 0.26 +/- 0.08 ng/ml on day 3, 0.19 +/- 0.07 ng/ml on day 5, 0.17 +/- 0.09 ng/ml on day 11, 0.11 +/- 0.02 ng/ml on days 15-30, and not detectable after 45 days of life. We also studied the relation between serum levels of DLIF and bilirubin in 23 jaundiced newborns between 3-5 days of life. We found a highly significant positive correlation between serum bilirubin concentrations and DLIF. These findings support the assumption that DLIF plays a role in impeding bilirubin excretion in the neonatal period, perhaps by inhibiting the activity of (Na-K)ATPase. PMID- 2545074 TI - Clinical manifestations of respiratory tract infections due to respiratory syncytial virus and rhinoviruses in hospitalized children. AB - From September 1984 to May 1986, nasopharyngeal secretions were obtained from 519 children with some form of respiratory tract infection. The nasal secretions were screened for respiratory syncytial virus (RSV), rhinoviruses, adenoviruses, parainfluenza virus types 1, 2, 3, influenza virus types A and B, and enteroviruses by tissue culture virus isolation technique and/or enzyme-linked immunosorbent assay. A uniform questionnaire gave information about age, sex, individual signs and symptoms, findings of the physical examination and clinical diagnosis of the patients. RSV was detected in 119 (23%) specimens and was thus the most frequent causative agent of respiratory infections. After RSV, rhinoviruses were the most frequently recovered pathogens accounting for 60 (12%) cases of acute respiratory disease. A comparison of the individual signs and symptoms, the findings of the physical examination and the clinical diagnosis of RSV and rhinovirus infected children revealed that there was no characteristic clinical pattern associated with either of the two viral respiratory pathogens. According to our results, rhinovirus infections were a major cause of lower respiratory tract infections in hospitalized children less than or equal to 3 years old. PMID- 2545075 TI - Response to growth hormone-releasing hormone as evidence of hypothalamic defect in optic nerve hypoplasia. AB - Hypothalamic-pituitary function was studied in four children with Optic Nerve Hypoplasia (ONH). All were found to be growth hormone deficient when provoked with glucagon or insulin induced hypoglycaemia (ITT), but did respond to bolus injection of GHRH. This indicates a primary hypothalamic defect. Virtual absence of pituitary tissue on high resolution CT scan explained the poor response of one child. One child has shown an excellent response to treatment with subcutaneous GHRH, which is physiologically the most appropriate treatment for this condition. PMID- 2545076 TI - Adrenocorticotropic hormone unresponsiveness associated with hypertrophic cardiomyopathy. AB - The clinical and autopsy findings in a case of adrenocorticotropic hormone unresponsiveness associated with hypertrophic cardiomyopathy are reported. A four month-old female with feeding difficulties and skin hyperpigmentation from two months of age was admitted with convulsions. She was hypoglycemic with normal serum electrolytes and the presence of hypertrophic cardiomyopathy was indicated by electrocardiogram and echocardiogram. Cardiac arrest occurred on the second hospital day. Low serum cortisol, high plasma adrenocorticotropic hormone, low urinary 17-ketosteroids, and normal urinary aldosterone excretion were documented after her death. Hypoplasia of the adrenal cortex with a persistent fetal zone and concentric hypertrophy of the heart were found at autopsy. We propose that the pathogenesis of this disease lies in impaired remodelling of the fetal adrenal cortex into the permanent cortex, and postulate an effect of adrenocorticotropic hormone on the myocardium as the cause of hypertrophic cardiomyopathy. PMID- 2545077 TI - Cytochemical studies of peripheral blood leucocytes in pregnancy. AB - Three enzymes (alkaline phosphatase, acid phosphatase and peroxidase) and two metabolites (glycogen and lipids), were studied in the circulating leucocytes of pregnant subjects attending the Antenatal Clinic, Ile-Ife State Hospital, during their second and third trimester of pregnancy. Values obtained were compared with those of a non-pregnant control group in the same locality. Leucocyte alkaline phosphatase, glycogen and lipid levels were found to be significantly elevated, while acid phosphatase could not be demonstrated in the study group. Peroxidase levels decreased in the second trimester, only to rise again in the third trimester. Peroxidase had a highly negative correlation with birth weight (r = 0.96) and was, therefore, suggested as a possible prognostic indicator of birth weight. Glycogen and lipids correlated significantly and positively with the Apgar score, and therefore could possibly be useful in antenatal assessment of foetal health status. The necessity for further investigations and confirmation of the possible clinical uses of these parameters in pregnancy has been highlighted. PMID- 2545078 TI - A study of chronic neurotic illness in Nigeria. AB - This is a report on a study of patients suffering from chronic neurotic illness, seen at the Psychiatric clinic of the University College Hospital, Ibadan. The patients' current mental health status was assessed using the General Health Questionnaire (GHQ) and also, the Special Neurotic Scale. Seventy-nine patients were studied, 28 males and 51 females. Seventy-one per cent of patients belong to the low socio-economic class, 49% had no formal education, and 38% had attended hospital for over 5 years. Patients who received diagnosis of anxiety neurosis generally did better than those with diagnosis of depressive neurosis (i.e. by their scores on the GHQ). Symptoms of psychological and sensory disturbance were most resistant to treatment. These and other findings are discussed. PMID- 2545079 TI - Hepatitis B surface antigen and immunoglobulin M complexes in chronic carriers and patients with acute and chronic liver diseases. AB - To determine the incidence and significance of circulating complexes between hepatitis B surface antigen (HBsAg) and immunoglobulin M (IgM), a total of 152 Nigerian adults with acute or chronic liver diseases were studied. They were compared with 100 chronic HBsAg carriers and 20 healthy volunteers. In HBsAg carriers HBsAg-IgM complexes persisted in 10% over a period of 6 months. In 53 cases of acute hepatitis B, 79% acute phase sera had detectable complexes. This initial incidence fell to 15% at 6 months. In the group of chronic liver diseases, the detection of HBsAg-IgM complexes occurred relatively infrequently. Circulating HBsAg-IgM complexes were associated with the presence of hepatitis B e antigen and a higher predisposition to chronicity after acute hepatitis. It is concluded that the presence of these complexes is specific to hepatitis B virus infections and may predict evolution to chronic liver diseases in Nigerians. PMID- 2545080 TI - Experimental infections of Plasmodium yoelii nigeriensis in mice and rats, and hosts' reactions. AB - In investigations into the course of infection of Plasmodium yoelii nigeriensis in rats and mice, it was found that adult mice, baby mice and baby rats were susceptible to the infection, while adult rats were totally refractive. The infection was rapidly fatal in baby and adult mice but was maintained with a low parasitaemia and tolerated by baby rats until the animals were 7 weeks old, after which they eradicated the infection totally. Although the adult rats did not exhibit any parasitaemia, there was a decrease in their haemoglobin and packed cell volume values. In adult mice, the haemoglobin and packed cell volume values were found to decrease with the increase in parasitaemia. PMID- 2545081 TI - Pyomyositis: a report on two cases from a region with a temperate climate. AB - Two cases of pyomyositis in Caucasians, from a region with temperate coastal climate is reported. The identification of staphylococci as an aetiological factor in both temperate and tropical regions is mentioned, and so are other possible identical aetiological factors. PMID- 2545082 TI - Infective factors of male infertility among Nigerians. AB - Seminal fluid from 782 Nigerian males with complaints of infertility were examined with respect to infective agents and indices such as sperm count, motility and the presence of a significant number of pus cells. Various infective agents were recovered from 54 (7%) of the patients, while in 25% of the remaining patients, a significant number of pus cells was present, with associated abnormal seminal fluid indices. Our findings indicate that seminal fluids constitute an important medium for the spread of various infective agents, and that genital infections by these infective agents, sexually and non-sexually transmitted, may be responsible for a good percentage of infertility cases in Nigerian males. PMID- 2545083 TI - Ultrasound measurement of foetal head to abdomen circumference ratio in pregnant Nigerians. AB - The ratio of foetal head to abdomen circumference ratio has been established in 242 foetuses scanned between 16 weeks and 40 weeks of gestation. The ratio was observed to fall gradually from 1.30 at 16 weeks to 1 at 30-32 weeks, and more slowly thereafter to 0.95 at term. The value of this ratio in differentiating between symmetrical and asymmetrical growth-retarded foetuses and in the management of such pregnancies is discussed. PMID- 2545084 TI - Morbidity pattern among Nigerian children from a poor urban community. AB - In a longitudinal study of the pattern of morbidity in a cohort of Nigerian children under five from a poor urban community, symptoms of respiratory tract infection were found to be the commonest during the study period. Nasal discharge had the highest incidence rate of 30.3 episodes per 100 child days followed by cough which had 12.2 episodes per 100 child days. Fever with or without other symptoms had an incidence of 10.8 episodes per 100 child days. The incidence of diarrhoeal disease of 3.3 episodes per 100 child days was much lower than that of the respiratory symptoms. When stratified by age, the respiratory symptoms and fever were common among the younger age groups with a peak incidence at 13-24 months. Diarrhoea on the other hand had an earlier peak than the other three symptoms, being most frequent among those aged 7-12 months. Skin rash had the highest incidence rate among children aged 37-60 months. Although it is not possible to describe a definite seasonal pattern of symptoms from this study, monthly distribution shows that nasal discharge occurred at an almost constant rate throughout the year, cough also occurred throughout the year with periods of decreased incidence in December and May. Fever as a symptom occurred throughout the study period with a decreased incidence in December and peaks in the late dry season. Although symptoms of diarrhoea were present throughout the study period, higher incidence rates occurred in the dry season. Skin rash occurred more frequently during the dry season and decreased during the rainy season. PMID- 2545085 TI - Specific protein pattern in adult healthy Nigerians. AB - Serum concentrations of total protein, albumin, total globulins, pre-albumin, alpha-1-antitrypsin, caeruloplasmin, transferrin, immunoglobulins (IgG, IgA, IgM) and plasma fibrinogen were determined in 120 healthy adult Igbo-Nigerian subjects (60 males and 60 females). There was no sex or age related differences in the concentration of the various specific proteins, although a significant sex difference was observed in the mean level of plasma fibrinogen. Our values for caeruloplasmin, transferrin and IgM were significantly higher, although IgG, IgA and plasma fibrinogen were lower than reported in previous studies on other ethnic groups of Nigerians. PMID- 2545086 TI - Non-specific bronchial airway hyperreactivity and the diagnosis of asthma. AB - In very mild and atypical cases of asthma, highly discriminative tests are needed to make the diagnosis. To demonstrate this, measurement of non-specific bronchial airway hyperreactivity by means of standardized bronchial inhalation challenge tests with histamine and methacholine were performed in 10 very mild asthmatic and nine normal control subjects; both groups included Nigerians who were temporarily resident in London at the time of the study. Bronchial reactivity was expressed as the provocative concentration of the agents causing a 20% fall in forced expiratory volume in 1 sec (PC20FEV1); higher values indicating lower levels of non-specific bronchial reactivity. The level of non-specific bronchial reactivity in these very mild asthmatics, whose baseline physiological data were not different from those in normals, was found to be 18-29 times higher than the normal control subjects. These tests very effectively discriminated between asthmatic and normal control subjects. With available resources it should be possible to study a large number of Nigerians in their own environment. PMID- 2545087 TI - An in-vitro study on ciprofloxacin and other anti-microbials against gram negative bacteria isolated from patients in Ibadan, Nigeria. AB - The high prevalence of common clinical isolates that are resistant to multiple antibiotics calls for regular review of the anti-microbial sensitivity pattern among bacteria of clinical significance in our environment. In the present study an increasing percentage of common isolates from hospitalized patients have been found to be resistant to Gentamicin and Cefotaxime, which play an important role in the chemotherapy of infections. Of special significance is the finding that over 60% of pseudomonads are now resistant to Gentamicin. The new fluoroquinolone, Ciprofloxacin, showed strong activity against all the isolates tested, with MIC values within the range of those reported as sensitive from many overseas centres. It should prove to be a valuable agent in the management of infections due to these organisms. PMID- 2545088 TI - A comparative study of the growth status of children with and without SS disease at Ilorin, Kwara State, Nigeria. AB - An anthropometric study was conducted to determine the growth status of 1698 children with and without SS disease. Of the total subjects, 979 (57.7%) were from the University of Ilorin elementary school. Children with SS disease made up 42.3% (719) of the total sample. Children with SS disease had anthropometric values which were lower than the 50th percentile of the Unilorin sample. Of all the parameters assessed, the weight-for-age for children with SS disease was particularly deficient compared with that of the Unilorin pupils and with the Harvard standard. Although subjects were from the same Ilorin urban areas, effort was made to match the socio-economic class of children with SS disease with the Unilorin children. The need for the prompt medical management of children with SS disease and the provision of health counselling services for them were recommended. PMID- 2545089 TI - Radioisotope investigations of haematological disorders (excluding sickle cell disease) in Sierra Leone. AB - Using 51Cr, 125I, and 59Fe, red cell survival, blood volumes and certain aspects of iron metabolism were investigated in Sierra Leoneans referred for further evaluation of various blood disorders. The data provided clarification of the nature of the anaemia in some patients and demonstrated the haemolytic and erythropoietic role of the spleen in others with splenomegaly. Blood volume values in healthy individuals were found to be similar to those of normal subjects in other populations. Increased plasma volumes commonly described in non African patients with anaemia and splenomegaly were also recorded in this group of Sierra Leoneans. Information obtained from these studies indicates that facilities for radioisotope investigations would be an asset in the diagnostic evaluation of complex blood disorders and in haematological research in developing African countries. PMID- 2545090 TI - Acute effect of cigarette smoking on peak expiratory flow rate and ventilation in resting Nigerian subjects. AB - The effects of cigarette smoking on peak expiratory flow rate (PEFR) and ventilation (V) in resting Nigerian subjects had been investigated in healthy male subjects (10 smokers, 8 non-smokers). The PEFR, V and tidal volume (VT) decreased significantly after smoking two medium-tar cigarettes. The action of smoke particles (carbon and tar) on the irritant receptor reflex of the lung, thus producing bronchoconstriction, and the depressant action of nicotine on the peripheral chemoreceptors, central chemoreceptors and respiratory centre may be the causal factors. PMID- 2545091 TI - Coronary microvascular narrowing in acute murine coxsackie B3 myocarditis. PMID- 2545092 TI - Changes in dyspnea-fatigue ratings as indicators of quality of life in the treatment of congestive heart failure. AB - A new clinical index of dyspnea and fatigue has been applied to rate the condition of patients with congestive heart failure. The index has 3 components, each rated on a scale from 0 to 4, for the magnitude of the task that evokes dyspnea or fatigue, the magnitude of the pace (or effort) with which the task is performed and the associated functional impairment in general activities. The ratings for each component are added to form an aggregated score, which can range from 0, for the worst condition, to 12, for the best. Because dyspnea and fatigue are prime symptoms and sources of clinical distress, the index helps reflect the quality of life in patients with congestive heart failure. In double-blind trials of therapy, changes in the index showed good correlations with patients' self selected ratings of improvement. The posttherapeutic changes in the index ratings were significantly higher with a new active agent (lisinopril) than with placebo or another active agent (captopril). PMID- 2545093 TI - Terminal ileitis associated with cytomegalovirus and the acquired immune deficiency syndrome. AB - A 41-yr-old homosexual male had cytomegalovirus infection localized to the terminal ileum as his initial and sole evident opportunistic infection due to the acquired immune deficiency syndrome. The symptoms, signs, roentgenographic findings, and appearance at surgery were suggestive of Crohn's disease. The pathogen was identified only by microscopic examination of bowel resected during a second laparotomy. The terminal ileum had profound mucosal ulceration and transmural fibrosis without granulomas. This novel report suggests that cytomegalovirus infection should be carefully searched for in immunocompromised patients presenting with clinical features suggesting Crohn's disease. PMID- 2545094 TI - Toxic megacolon in cytomegalovirus colitis. AB - We report a case of toxic megacolon manifesting in cytomegalovirus (CMV) colitis in a 55-yr-old man with steroid-dependent chronic obstructive pulmonary disease. He presented to the hospital with increasing dyspnea and low-grade fever. His hospital course was characterized by the poor response of his symptoms to treatment, and by the subsequent development of intermittent hematochezia and, eventually, acute abdomen. The surgical specimen showed dilatation of the cecum and ascending colon with a solitary mucosal ulcer in the latter. The major histologic changes were limited to the area of ulceration. In addition to classical CMV inclusions. vasculitis manifested in two forms, namely, leukocytoclastic type and fibrinoid necrosis. The patient died shortly thereafter, due to multi-organ system failure. To our knowledge, this represents the first reported case of toxic megacolon due to CMV infection without underlying inflammatory bowel disease. The pathogenesis of toxic colonic dilatation remains unknown. PMID- 2545095 TI - Difficult preoperative diagnosis of a patient with sclerosing splenic hemangioma. AB - We present a young asymptomatic woman with splenomegaly and a large isolated splenic mass demonstrated by ultrasonography, 99mTc sulfur colloid, and gallium scintigraphy studies. Computerized tomography (CT) and three-phase 99mTc-labeled red blood cell imaging suggested a malignant lesion. Repeated sonographically guided fine needle aspiration (FNA) obtained only blood, suggesting the possible vascular nature of the tumor. Splenectomy established the diagnosis of splenic hemangioma (SH) with marked sclerotic changes. We conclude from this case that 1) the sclerotic and cystic changes in the SH and the abdominal lymphadenopathy could explain why the three-phase red blood cell and CT scanning, respectively, suggested that the lesion was malignant rather than benign; 2) guided FNA of a splenic mass suspected to be hemangioma may be an additional safe and useful diagnostic procedure. Multiple aspirations yielding blood alone suggest hemangioma and may prevent an unnecessary operation. To the best of our knowledge, this is the first reported case in the literature of FNA of splenic hemangioma. PMID- 2545097 TI - Crystalline glomerular inclusions in multiple myeloma. AB - Prominent crystalline inclusions were noted in the glomeruli of a 57-year-old man with a 6-month history of swelling and pain in the hands, slight proteinuria, and elevated serum creatinine and BUN. The crystalline inclusions were most prominent in the visceral epithelium but were also noted in endothelial and mesangial cells, in the parietal epithelium, and in the epithelial cells of the proximal tubules. On electron microscopy the crystalline structures were of various geometric shapes but had no definite substructure. Immunofluorescence was negative. The patient was considered to have a hitherto undescribed metabolic disease. Several months later the patient underwent an operation for carpal tunnel syndrome and amyloid deposits, crystalline inclusions similar to those noted in the kidney were observed in the synovial tissue. Shortly after this the patient was found to have multiple myeloma of IgG, kappa type with Bence Jones proteinuria. Lymphoplasmacytic cells in the bone marrow contained the same crystalline inclusions noted in the kidney and synovium. This case therefore seems to represent a new and very rare form of glomerular involvement in multiple myeloma. PMID- 2545096 TI - Diet, mammographic features of breast tissue, and breast cancer risk. AB - This case-control study was designed to reevaluate the association of the morphology of breast tissue seen on mammograms with breast cancer risk and to assess the relation of diet, especially intake of fat and vitamin A, to the high risk mammographic images. The cases included 290 patients with newly diagnosed breast cancer who were first treated in Quebec in 1982-1984. The controls included 645 women who participated in the Canadian National Breast Screening Study. Risk of breast cancer was higher among women with the P2 or DY parenchymal pattern (relative risk (RR) = 3.7, 95% confidence interval (Cl) 2.0-7.0) than it was among those with the N1 pattern. Moreover, risk increased regularly with the extent of nodular and homogeneous densities on the mammogram. Relative risk was 5.5 (95% Cl 2.3-13.2) for women in whom 60% or more of the volume of the breast showed either nodular or homogeneous densities compared with women without such densities. Among controls, increase in energy-adjusted saturated fat intake was associated with an increase in extent of high-risk mammographic features. Energy adjusted polyunsaturated fat or cholesterol intake did not, however, appear to influence the morphology of breast tissue seen on the mammogram. Increasing carotenoid and fiber intakes were associated with a reduction of the extent of densities on the mammogram, but retinol intake seemed to have little or no effect on mammographic features. These data suggest that elevation in saturated fat intake and reduction in carotenoid and fiber intakes may be related to an increase in breast cancer risk through effects of these nutrients on breast tissue morphology. PMID- 2545099 TI - Fatal hepatic necrosis caused by disseminated type 5 adenovirus infection in a renal transplant recipient. AB - This is an unusual case of fatal hepatic necrosis caused by disseminated type 5 adenovirus in a renal transplant recipient. This adult patient may have been colonized at the time of transplantation with a kidney from a pediatric donor. The adenovirus became invasive when the host's cellular immune system was suppressed by high doses of corticosteroids given to reverse acute allograft rejection. PMID- 2545098 TI - Distal deletion of chromosome Ip in ductal carcinoma of the breast. AB - By use of recombinant DNA probes that correspond to genetic loci residing on human chromosome 1, DNA samples from 37 ductal breast carcinomas and constitutional DNA from the same individuals were tested for loss of heterozygosity. A high frequency (41%) of reduction to homozygosity was detected with the probe p1-79, which recognizes the highly polymorphic locus D1Z2, localized on 1p36. Loss of heterozygosity at other chromosome 1 loci was much less common, not exceeding a frequency of 10%, and was never observed in the absence of the D1Z2 loss. Somatic loss of heterozygosity at D1Z2 was more frequent in patients with a strong family history of breast cancer (60%), in patients with early diagnosis (before 45 years of age) (70%), and in those with multiple tumors or tumor foci (50%) than in patients with none of the characteristics of hereditary tumors (21%). No associations were observed between loss of heterozygosity and prognostic factors. These results suggest that inactivation of a tumor suppressor gene located on the distal portion of chromosome 1p, alone or combined with other genetic changes, may represent a fundamental step in the pathogenesis of ductal carcinoma of the breast. PMID- 2545100 TI - Experimental studies on the maintenance of Toscana and Arbia viruses (Bunyaviridae: Phlebovirus). AB - Transovarial transmission (TOT) of Toscana (TOS) and Arbia (ARB) viruses in a laboratory colony of Phlebotomus perniciosus is reported. Toscana and ARB viruses were maintained in P. perniciosus females, initially infected by intrathoracic inoculation, for 2 and 3 consecutive generations respectively. TOT was demonstrated in F1 (75%) and F2 (67%) generation adults for TOS and F1 (47%), F2 (37%), and F3 (34%) generation adults for ARB virus. The progressive decline of virus infection rates in each generation suggests that these agents cannot be maintained indefinitely by TOT. No infection was observed in F1 progeny after female parents were fed through membranes with either virus. Transovarially infected females were able to transmit TOS virus by bite to a susceptible vertebrate. Venereal infection of P. perniciosus females mated to males transovarially infected with TOS virus was seen. PMID- 2545101 TI - Intraoperative ultrasonography in resection of small hepatocellular carcinoma associated with cirrhosis. AB - Preoperative detectability rates of hepatocellular carcinoma smaller than 5 cm in 113 cirrhotic patients were 91 percent by ultrasonography, 93 percent by computed tomography, and 92 percent by selective angiography. The combination of two methods produced detectability rates of 97 to 99 percent. One hundred three patients underwent various types of hepatic resection with the aid of intraoperative ultrasonography. Forty-four tumors (43 percent) were embedded in the liver, and these tumors were not detected by conventional surgical exploration. The detectability rates were 38 percent for hepatocellular carcinomas smaller than 2 cm, 57 percent for 2 to 3.5 cm tumors, and 71 percent for 3.5 to 5 cm tumors. All undetectable hepatocellular carcinomas were identified by intraoperative echography. The overall detection rate by this method was 98 percent, which was substantially higher than the preoperative rate. Intraoperative ultrasonography is a useful and indispensable method for performing atypical minor hepatectomy for the treatment of small hepatocellular carcinomas associated with liver cirrhosis. PMID- 2545103 TI - [The kallikrein-kinin system of the blood in gestosis]. AB - Blood kallikrein-kinin system (KKS) was studied in 80 pregnant women with gestoses of various severity and in 20 females with normal pregnancy. Kinin formation and degradation were analysed and the exhaustion of these processes stated. Its degree was found in proportion with gestosis severity. Prekallikrein levels depended on the predominance of hypertensive disease or albuminuria occurrence. The aforementioned exhaustion possibly resulted from the preliminary activation of kallikrein-kinin system links which took place at the initial stages of gestosis to promote the adjustment of female circulation to increased loads. PMID- 2545102 TI - Role of polymorphonuclear leukocytes in collagenase production in chronic otitis media. AB - The effect of polymorphonuclear leukocytes on collagenase production by fibroblast-like cells was studied in an in vitro cell culture system. An extract of rat intraperitoneal polymorphonuclear leukocytes induced significant collagenase production by rat skin fibroblast-like cells. Fibroblast-like cells cultured in the presence of polymorphonuclear leukocytes had many conspicuous microspikes on their surface and contained bundles of actin filaments in their cytoplasm. Polymorphonuclear leukocytes appears to be involved in bone resorption by inducing collagenase production by fibroblast-like cells, as well as by releasing their own enzymes involved in bone resorption. PMID- 2545104 TI - [Urinary function in women with uterine myoma]. PMID- 2545105 TI - Strong-cation-exchange sulfoethyl aspartamide chromatography for peptide mapping of Staphylococcus aureus V8 protein digests. AB - In two recent reports (D. L. Crimmins, J. Gorka, R. S. Thoma, and B. D. Schwartz (1988) J. Chromatogr. 443, 63-71; A. J. Alpert and P. C. Andrews (1988) J. Chromatogr. 443, 85-96) a sulfoethyl aspartamide column was shown to efficiently analyze peptides less than 25 residues in length which differ in the number of nominal positive charges at pH 3.0. In particular, the elution order for a series of distinct peptides ranging in nominal charge from +1 to +7 was found to be monotonic in nature indicating that separation was primarily via a cation exchange mechanism. The present study employs this chromatographic system to isolate and characterize major fragments of proteolytic digests. Six commercially available proteins of known sequence (myoglobin, beta-casein, concanavalin A, carbonic anhydrase, lentil lectin, and enolase) were digested with Staphylococcus aureus V8 to generate peptide fragments. The resulting mixture was chromatographed on a sulfoethyl aspartamide column to isolate major fragments which were then subjected to amino acid analysis and N-terminal sequencing. With complete proteolysis (i.e., peptide fragments terminating in either an aspartic or a glutamic acid) separation of the fragments should result from the sum of histidine, lysine, and arginine residues contained in each fragment. Most of the peptide fragments eluted at the expected time on the sulfoethyl aspartamide column. Those fragments with anomalous behavior resulted from incomplete cleavage or cleavage at nonacidic residues or were greater than 35 residues in length. Each proteolytic digest was also analyzed by standard reverse-phase C4 chromatography to compare the peptide maps for these two distinct chromatographic modes. PMID- 2545106 TI - Determination of the A-T content of DNA by second-derivative ultraviolet spectroscopy. AB - A method for the determination of the A-T content of DNA based on second derivative ultraviolet spectra is presented. It allows measurement in a wide range of pH values, ionic strengths, and buffer media. It is nondestructive for the sample and requires not more than 10 micrograms of DNA. PMID- 2545107 TI - Radioactive-electrophoretic assay of adenosine 5'-triphosphate sulfurylase activity in crude extracts with sulfate or selenate as a substrate. AB - An assay method for ATP sulfurylase is presented which employs Na2(35)SO4 as a substrate and measures the production of labeled adenosine 5'-phosphosulfate and 3'-phosphoadenosine 5'-phosphosulfate by low-voltage, hanging paper strip electrophoresis. The method is applicable to crude bacterial or mammalian extracts and accurately measures picomole amounts of product(s). Na2(75SeO4 can also be employed as a substrate, if the unstable radioactive product, adenosine 5'-phosphoselenate, is converted to elemental 75Se degrees by inclusion of reduced glutathione in the reaction mixture. The same paper strip electrophoretic technique can then be used to separate 75Se degrees from the radiolabeled substrate. The method also has utility for measuring any direct reduction by crude microbial extracts of radioactive selenate to selenite, independent of ATP sulfurylase. PMID- 2545108 TI - Separation of hepatocyte plasma membrane domains by free flow electrophoresis. AB - We have applied free flow electrophoresis to separate the canalicular and basolateral (sinusoidal and lateral) domains of rat hepatocyte plasma membranes. Hepatocyte plasma membranes were prepurified by rat zonal and discontinous sucrose gradient centrifugation. In electrophoretic separation, the canalicular membranes were more deflected toward the anode than the basolateral membranes. Na+-dependent taurocholate uptake could be measured in both membrane fractions, transport activity being highest in fractions containing the highest specific activity in the basolateral marker enzyme Na+-K+-ATPase. Thus, differences in electrophoretic mobility permit the separation of functional intact plasma membrane vesicles derived from basolateral and canalicular plasma membrane domains of rat hepatocyte. PMID- 2545109 TI - A new method of plasmid DNA preparation by sucrose-mediated detergent lysis from Escherichia coli (gram-negative) and Staphylococcus aureus (gram-positive). AB - A simple and cheap method of plasmid DNA preparation from both gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) organism is presented here. In this method, in place of the high-priced chemicals lysostaphin and lysozyme which are commonly used for removal of cell-wall during plasmid DNA preparation from gram-positive and gram-negative bacteria, respectively, only sucrose has been used. Firstly, bacteria is treated with Trizma (pH 8.0) containing 100% sucrose (hypertonic solution). Due to this osmotic shock, protoplasm covered by the plasma membrane of bacteria possibly shrinks and becomes detached from the cell-wall. Osmotically sensitive cells thus formed, from gram-positive (S. aureus) and gram-negative (E. coli) bacteria, are finally lysed by the lysis mixture, containing brij 58 and sodium deoxycholate. The lysate is centrifuged at 15,000 rpm for 30 min to pellet the cell debris. The supernatant containing plasmid DNA is treated with either polyethylene glycol or isopropanol. The precipitate which contains plasmid DNA is dissolved in a buffer containing Tris, EDTA, NaCl, and sodium dodecyl sulfate (pH 8.0); thus protein is denatured and removed. Finally, RNA is removed by RNase treatment. The average yield of staphylococcal plasmid DNA as well as plasmid pBR322 from E. coli HB101 in 100% sucrose-treated preparations is greater than that of lysostaphin- and lysozyme-treated preparations. This method is applicable for both large-scale and small-scale preparations. The substrate activity for restriction enzyme, cloning, transforming ability, and electron microscopic profile of the plasmid DNA prepared by this method remains unaltered. PMID- 2545110 TI - Glycerol, sodium phosphate, and sodium chloride permit the solubilization and partial purification of rat hepatic alpha 1-receptors by 3-(3-cholamidylpropyl) dimethylammonio-1-propanesulfonate. AB - CHAPS [3-(3-cholamidylpropyl)-dimethylammonio-1-propanesulfonate], a zwitterionic detergent, has been used to solubilize the rat hepatic alpha 1-adrenergic receptor. Although the use of this detergent alone permitted a poor receptor solubilization, the inclusion of sodium phosphate, sodium chloride, and glycerol to the medium allowed 30% of the binding activity observed in plasma membranes to be recovered. Binding of the selective alpha 1-adrenergic antagonist, [3H]prazosin, by the solubilized preparation was saturable and of high affinity. In addition, binding of the radioligand was inhibited by a variety of adrenergic agents with affinity, specificity, and stereoselectivity comparable to that observed in plasma membranes. The use of glycerol in the solubilization medium permitted recovery of the solubilized receptor in a stable form (T1/2 = 72 h at 4 degrees C). Sequential affinity and size-exclusion gel chromatography allowed a 1000-fold purification of the solubilized receptor. The Stokes' radius and the apparent molecular mass of the purified receptor-Chaps complex (48.4 A and 160,000 Da, respectively), determined by gel filtration chromatography, were similar to those previously obtained for the rat hepatic alpha 1-receptor purified after solubilization with the nonionic detergent digitonin. These data indicate that the combination of Chaps, sodium phosphate, sodium chloride, and glycerol permitted the solubilization and partial purification of hepatic alpha 1 receptor in an active and stable form. The use of this technique might be useful for the solubilization of other membrane-bound proteins by Chaps whose biophysical characteristics make it an ideal detergent for reconstitution experiments. PMID- 2545111 TI - Determination of [U-13C]glucose turnover into various metabolite pools for the differential diagnosis of lactic acidemias. AB - Stable-isotope tracer experiments performed in vitro are evaluated for their utility in differentiating between pyruvate dehydrogenase and cytochrome oxidase deficiencies, two of several enzyme defects commonly associated with the lactic acidemias. Fibroblasts of enzyme-deficient individuals and of age-matched controls are grown in medium containing [U-13C]glucose. Direct analysis of cells and conditioned culture medium provides only minor differences in the organic acid/amino acid GC-MS profiles, making differentiation of enzyme defects difficult by this method. However, differences have been found in the glucose turnover into various cell metabolites, making differentiation of these two enzyme defects possible. The cellular pool of glutamic acid experiences 13C enrichment in both the control and cytochrome oxidase deficient lines, but not in the pyruvate dehydrogenase-deficient line. The cellular pool of an unknown, possibly an aminopentose sugar, on the other hand, experiences 13C-enrichment in the pyruvate dehydrogenase and control lines, but not in the cytochrome oxidase line. These observations, as well as other differences in the extent of enrichment into various metabolite pools, suggest that this stable-isotope approach, in vitro, is feasible and may allow these two enzyme defects to be differentiated in a definitive manner. Such stable-isotope experiments are easy to carry out with cultured cells and are inexpensive. Applications of the technique to other genetic disorders might be appropriate. PMID- 2545112 TI - A simple colorimetric assay method for pyrophosphate in the presence of a 1000 fold excess of orthophosphate: application of the method to the study of pyrophosphate metabolism in mitochondria. AB - A sensitive colorimetric method for the assay of inorganic pyrophosphate with excess of orthophosphate is described. The principle of this method lies in the formation of phosphomolybdate and PPi-molybdate complexes with subsequent extraction of the phosphomolybdate complex by organic solvents and reduction of the PPi-molybdate complex by dithiothreitol and Eikonogen. The sensitivity of the method was from 5 to 120 nmol of PPi in a 2.0-ml sample. PMID- 2545113 TI - High-performance liquid chromatography determination of enzyme activities in the presence of small amounts of product. AB - Enzymes can be assayed by HPLC by calculating the amount of substrate(s) left over, or product formed, through the peak area ratios with a suitable internal standard. However, sometimes the substrates used are contaminated with small amounts of products and this can lead to errors in the determination of the enzyme activity. A method for a HPLC test of such enzymes, which prevents eventual errors, uses the ratio substrate/product at time zero as internal standard and the kinetics can be followed with the aid of a simple mathematical equation. This approach was applied to the determination of the activities of papain, urokinase, NAD glycohydrolase, and pyruvate kinase samples and it was compared with the data obtained by the internal standard method, giving reproducible results in all cases. PMID- 2545114 TI - Problems associated with spin trapping oxygen-centered free radicals in biological systems. PMID- 2545115 TI - Determination of inositol 1,4,5-trisphosphate levels in Dictyostelium by isotope dilution assay. AB - A commercial isotope dilution assay was used for the determination of Ins(1,4,5)P3 levels in the microorganism Dictyostelium discoideum. Cross reactivity in the assay was detected with extracts from cells and the medium. The compound which induced this cross-reactivity was tentatively identified as Ins(1,4,5)P3 by (i) codegradation with authentic [32P]Ins(1,4,5)P3 by three specific Ins(1,4,5)P3 phosphatases, and (ii) co-chromatography with authentic [32P]Ins(1,4,5)P3 on HPLC columns. The cellular concentration was estimated as 165 +/- 42 pmol/10(8) cells, yielding a mean intracellular Ins(1,4,5)P3 concentration of 3.3 microM. Dictyostelium cells secrete large amounts of Ins(1,4,5)P3 at a rate of about 10% of the cellular content per minute, yielding about 0.13 microM extracellular Ins(1,4,5)P3 after 15 min in a suspension of 10(8) cells/ml. The chemoattractant cAMP induced a transient increase of the Ins(1,4,5)P3 concentration; the data suggest an intracacellular rise from 3.3 to 5.5 microM with a maximum at 6 s after stimulation. PMID- 2545116 TI - Simultaneous determination of iodate and periodate by capillary zone electrophoresis: application to carbohydrate analysis. AB - Iodate and periodate were rapidly (in 11 min) separated from each other with high column efficiency by capillary zone electrophoresis, using a fused silica tube (50 microns i.d., 80 cm) and 100 mM acetate buffer, pH 4.5, as carrier. On-column uv detection at 222 nm allowed sensitive detection down to the picomole level, and measurement of relative peak area to that of pyromellitic acid (internal standard) enabled reproducible determination of these ions. This method was proved useful for periodate oxidation analysis of various carbohydrates. PMID- 2545117 TI - Synthesis and properties of a nonhydrolyzable adenosine phosphosulfate analog. AB - Initial activation of inorganic sulfate for subsequent synthesis of sulfated biomolecules requires the action of ATP-sulfurylase to generate adenosine 5' phosphosulfate (APS). This activated sulfate intermediate is both chemically labile and susceptible to enzymatic degradation. Consequently, it has not proven useful as a ligand for either purification or characterization of the various APS utilizing enzymes. For these purposes, a stable analog of APS was required. This paper describes the simple and efficient synthesis and structural confirmation of a nonhydrolyzable APS analog, beta-methylene APS, with an overall molar yield of 40-50%. The method involves nucleophilic substitution of the chlorine moiety of a 5'-chloromethylphosphonate ester of 2',3'-O-isopropylidene adenosine by a sulfite ion. We also report the initial utilization of this compound as an inhibitor in kinetic trials of both ATP-sulfurylase and APS kinase and as an affinity ligand for the purification of these two APS-utilizing enzymes from cartilaginous tissue. PMID- 2545118 TI - Multiple nerve palsies after anesthesia--a complication of nitrous oxide use? PMID- 2545119 TI - Outbreak of nosocomial Flavobacterium meningosepticum respiratory infections associated with use of aerosolized polymyxin B. AB - Flavobacterium meningosepticum is an uncommon cause of adult nosocomial infection. On a medical/surgical intensive care unit we recently encountered an adult outbreak of respiratory colonization and infection caused by this organism, which was associated with the prophylactic use of aerosolized polymyxin B that had been used in an attempt to abort an outbreak of infection caused by highly resistant strains of Pseudomonas aeruginosa. Twenty isolates (95% from respiratory secretions) of F. meningosepticum from nine persons were identified during a 2 1/2-month period. No environmental source has been identified to date. Pneumonia developed in five patients, and two deaths associated with this organism occurred. All isolates were sensitive to ciprofloxacin; none were sensitive to other antibiotics tested, including third-generation cephalosporins, aminoglycosides, erythromycin, trimethoprim-sulfamethoxazole, antipseudomonal penicillins, aztreonam, and imipenem/cilastatin. Two patients with nosocomial pneumonia were successfully treated with oral ciprofloxacin. F. meningosepticum may emerge as an important pathogen if prophylactic use of polymyxin B becomes more widespread. Ciprofloxacin may become the agent of choice for treatment of this organism. PMID- 2545121 TI - Bilateral breast cancer. AB - The magnitude of the clinical problem of bilateral occurrence of breast cancer is controversial. In order to evaluate clinical and pathologic factors associated with increased risk for metachronous, contralateral breast cancer, we reviewed the records of 187 patients undergoing operative treatment for primary breast cancer by the same surgeon at a single institution. Variables analyzed included age, race, history of tuberculosis or cancer at other sites, family history of breast cancer or cancer at other sites, prior use of estrogens, tobacco, and alcohol, marital status, parity, age at first pregnancy, tumor size and location, histologic diagnosis, degree of cellular differentiation and involvement of axillary lymph nodes. The incidence of metachronous, contralateral breast cancer was 11.8 per cent. The only factor with a statistically significant association with bilateral cancer was histologic diagnosis characterized by multicentricity. Life table survival analysis revealed that the five-year survival following treatment for metachronous, contralateral cancer was 59 per cent. We conclude that bilateral breast cancer is a significant clinical problem, that histology characterized by multicentricity is associated with a higher incidence of bilateral breast cancer, and that favorable survival justifies an aggressive approach. PMID- 2545120 TI - Condylomatous lesion (HPV infection) of uterine cervix. A clinical, colposcopic, cyto, histomorphological and immunohistochemical study. AB - 43, patients, subjected to cervical cytology, colposcopy and colposcopic directed biopsy, were found to have lesions suggestive of human Papillomavirus (HPV) infection by one or more parameters. Immunohistochemical staining for HPV antigen yielded positive results in 23 (53.5 p. cent) cases. Amongst 30 cases with HPV lesions both in cytology and histopathology, 19 (63.3 p. cent) were positive for the HPV antigen. This was significantly higher as compared to the groups where HPV lesion was observed in cytology or histology alone (p = 0.05). The rate of positivity was the highest (81.3 p. cent) in a group of 16 patients having low grade dysplasia (mild to moderate) and both cytology and histologic evidence of koilocytotic changes. Presence or absence of HPV lesion by colposcopic examination in these 43 cases did not make any significant difference in rate of antigen positivity. PMID- 2545122 TI - [Diabetic ketoacidosis in childhood: the first day of treatment]. AB - Fifteen patients with diabetic ketoacidosis treated at ICU in the last eight years with low-dose intravenous insulin infusion are retrospectively revised. Diabetic ketoacidosis was the initial presentation of diabetes in 12 children. Mild infections were the usual starting factor. Only the children with shock of blood pH less than or equal to 7.10 on admission received sodium bicarbonate. The average of pH correction was 6.4 h. The average of glycemia correction (less than or equal to 250 mg/dl) was 5 h with a rate of 120 mg/dl/h. None of children had complications in the course of the treatment, except for one of them who presented cerebral edema. This procedure is a safe and efficacious treated for diabetic ketoacidosis in children. PMID- 2545123 TI - [Simultaneous bilateral Wilms tumor]. AB - Bilaterality is uncommon in Wilms' tumor, being present in 4% to 8% of the cases. Definite progress has been made in the treatment with marked improvement in prognosis. The recent trend toward more conservative sugery, preoperative chemotherapy an avoidance of high-dose radiation therapy has yielded good results. This is confirmed in our two cases with synchronous tumors. PMID- 2545124 TI - Persistent Cryptococcus neoformans infection of the prostate after successful treatment of meningitis. California Collaborative Treatment Group. AB - STUDY OBJECTIVE: To assess the frequency of persistent Cryptococcus neoformans infection in patients with the acquired immunodeficiency syndrome (AIDS) after receiving apparently adequate treatment for meningitis. DESIGN: Blood, urine, and cerebrospinal fluid were cultured at the conclusion of primary therapy to assess the adequacy of treatment. SETTING: Outpatient clinics at three medical centers. PATIENTS: Patients had C. neoformans grown in culture from cerebrospinal fluid. Primary therapy consisted of either 2.0 g of amphotericin B alone; 6 weeks of combination therapy with flucytosine; or, if flucytosine was poorly tolerated, an adjusted minimum total amphotericin B dose. To meet criteria for adequate treatment of meningitis all patients had two sequential cerebrospinal fluid samples which were culture negative. MEASUREMENTS AND MAIN RESULTS: Nine of forty one patients grew C. neoformans from urine after completion of primary treatment, but none had urinary symptoms. Fungi were visualized in expressed prostatic secretions in 4 of these patients. One patient refused further treatment and developed cryptococcemia within 5 weeks. Three patients received additional amphotericin B; all had persistent funguria without systemic relapse. Six patients received fluconazole; 4 became urine culture negative, and 2 had systemic relapse. CONCLUSION: The persistence of urinary C. neoformans after adequate therapy for meningitis suggests that the urinary tract (probably the prostate) is a sequestered reservoir of infection from which systemic relapse may occur. PMID- 2545125 TI - General recommendations on immunization. Guidelines from the Immunization Practices Advisory Committee. Centers for Disease Control. AB - This revision of the "General Recommendations on Immunization" updates the 1983 statement. Changes or new sections include 1) listing of vaccines available in the United States by type and recommended routes, 2) updated schedules for immunizing infants and children, 3) clarification of the guidelines for spacing administration of immune globulin preparations and different vaccines, 4) an updated table of recommendations for routine immunization of children infected with human immunodeficiency virus, 5) listing of conditions that are often inappropriately considered contraindications to immunizations, and 6) addition of information on the National Childhood Vaccine Injury Act of 1986 and the National Vaccine Injury Compensation Program. These recommendations are not comprehensive for each vaccine: Immunization Practices Advisory Committee (ACIP) recommendations on each vaccine should be consulted for more details. PMID- 2545127 TI - Calcium channels: structure and function. PMID- 2545126 TI - Molecular properties of dihydropyridine-sensitive calcium channels. PMID- 2545128 TI - Voltage-dependent calcium conductances in mammalian neurons. The P channel. PMID- 2545129 TI - Structure, function, and regulation of the skeletal muscle dihydropyridine receptor. PMID- 2545130 TI - Biochemistry, molecular pharmacology, and functional control of Ca2+ channels. PMID- 2545132 TI - Modulation of calcium channels by charged and neutral dihydropyridines. PMID- 2545131 TI - Reconstitution of cardiac sarcoplasmic reticulum calcium channels. PMID- 2545133 TI - Structure of calcium channels. PMID- 2545134 TI - Calcium channels in smooth muscle. Properties and regulation. PMID- 2545135 TI - The inhibitory effects of omega-conotoxins on Ca channels and synapses. AB - Omega conotoxins are peptides from snail venom. Two variants, omega CgTX and omega CmTX derived from two species of Conus, are the subjects of this report. Part I of this report reviews and discusses the ability of these toxins to inhibit Ca channels and synapses in different tissues from various species of animals. The potencies of these toxins vary depending on the target tissue, consonant with the notion that synaptic Ca channels have changed in the course of evolution. Part II introduces the notion that in contrast to inorganic Ca channel blockers, which act by reducing the amount of Ca2+ ions that can permeate an open channel, omega toxins act by reducing the availability of functional Ca channels. Thus, Ca channel-inhibition by omega toxins and that by inorganic blockers are expected to produce qualitatively different alterations in the distribution of intracellular Ca2+. Consistent with this expectation, the dose-response curves of inorganic blockers and omega CmTX differ. The dose-response curves of inorganic blockers are thought to reflect the cooperativity of Ca2+ ions in mediating transmitter release. In contrast, comparison of experimental and theoretical dose response curves of omega CmTX leads us to propose the hypothesis that Ca channels normally do not act cooperatively to effect transmitter release. PMID- 2545136 TI - Voltage-operated calcium-channel subtypes in human neuroblastoma and rat pheochromocytoma cells. PMID- 2545137 TI - 32,000-Dalton subunit of the 1,4-dihydropyridine receptor. AB - Polyclonal antibodies to the 32,000-Da polypeptide of the 1,4-dihydropyridine receptor of the voltage-dependent Ca2+ channel have been produced and used to characterize the association of the 32,000-Da polypeptide (gamma subunit) with other subunits of the dihydropyridine receptor. The 32,000-Da polypeptide was found to copurify with alpha 1 and alpha 2 subunits at each step of the purification of the dihydropyridine receptor. Monoclonal antibodies against the alpha 1 and beta subunits immunoprecipitate the digitonin-solubilized dihydropyridine receptor as a multisubunit complex that includes the 32,000-Da polypeptide. Polyclonal antibodies generated against both the nonreduced and reduced forms of the alpha 2 subunit and the gamma subunit have been used to show that the 32,000-Da polypeptide is not a proteolytic fragment of a larger component of the dihydropyridine receptor and not disulfide linked to the alpha 2 subunit. Our results demonstrate that the 32,000-Da polypeptide (gamma subunit) is an integral and distinct component of the dihydropyridine receptor. PMID- 2545138 TI - Monoclonal antibodies against calcium channels. PMID- 2545139 TI - Antibodies against calcium channels in the Lambert-Eaton myasthenic syndrome. PMID- 2545140 TI - L-type calcium channels in cardiac and skeletal muscle. Purification and phosphorylation. PMID- 2545141 TI - Motor nerve terminal calcium channels in Lambert-Eaton myasthenic syndrome. Morphologic evidence for depletion and that the depletion is mediated by autoantibodies. PMID- 2545142 TI - Modulation of Ca channels in peripheral neurons. PMID- 2545143 TI - Calcium channels in vertebrate neurons. Experiments on a neuroblastoma hybrid model. PMID- 2545144 TI - Modulation of Ca2+-channel currents in sensory neurons by pertussis toxin sensitive G-proteins. PMID- 2545146 TI - Calcium currents in insulin-secreting beta-cells. PMID- 2545145 TI - Solubilization, partial purification, and properties of omega-conotoxin receptors associated with voltage-dependent calcium channels from rat brain synaptosomes. AB - These experiments provide a starting point for biochemical characterization of Ca channels from neuronal membranes, using omega-CgTX as a specific marker. The purification of the omega-CgTX receptors is far from complete. Each of the purification steps described results in only a two- to fivefold enrichment of the receptor proteins, and is accompanied by a loss of receptor concentration and stability, so the maximal specific activity achieved by a combination of these steps falls several orders of magnitude short of that of a large, homogeneous, active protein. Nevertheless, these studies have yielded important information about the omega-CgTX receptor. The Stokes' radius, determined from gel exclusion chromatography, is approximately 87 A, and the sedimentation coefficient, determined from sucrose gradient sedimentation, is approximately 19 S. These values are similar to those found for the DHP receptors solubilized in digitonin. We have also found that at least some of the omega-CgTX receptors have complex carbohydrate moieties that are recognized by WGA, together with evidence of heterogeneity of receptor glycosylation. Additionally, we have been able to use the solubilized, partially purified receptors in cross-linking experiments to tentatively identify the molecular weights of the omega-CgTX targets from rat brain. A large peptide of approximately 300 kDa, similar to that identified in photoaffinity studies, is very clearly labeled by the chemical incorporation of [125I]omega-CgTX into partially purified receptor preparations, but some ambiguity remains because of the faint labeling of peptides in the 120-170-kDa range. The approximately 300-kDa peptide is much larger than any single peptide component of DHP receptors from skeletal muscle, and it may be related to a molecular combination of the 170-kDa and 135-kDa subunits of the DHP receptor. Because [125I]omega-CgTX presumably labels both N- and L-type neuronal Ca channels, both channel types will probably be found in the purified preparations. Thus, at some time, it will be necessary to separate DHP-sensitive L-type channels from preparations of L- and N-type channels identified by omega-CgTX binding. PMID- 2545147 TI - Single calcium channel activity in mouse pancreatic beta-cells. PMID- 2545148 TI - Functions for calcium channels in pituitary cells. PMID- 2545149 TI - Appearance and function of voltage-dependent Ca2+ channels during pre- and postnatal development of cardiac and skeletal muscles. PMID- 2545150 TI - Properties of the calcium channel associated omega-conotoxin receptor in rat brain. PMID- 2545151 TI - Gating and permeation of different types of Ca channels. PMID- 2545153 TI - Block of Na+ ion permeation and selectivity of Ca channels. PMID- 2545152 TI - Mechanisms of interaction of permeant ions and protons with dihydropyridine sensitive calcium channels. PMID- 2545154 TI - Phencyclidine during pregnancy: behavioral and neurochemical effects in the offspring. PMID- 2545155 TI - [Warts and epidermoid carcinoma after renal transplantation]. AB - Kidney transplant recipients suffer in the long-term from several cutaneous disorders linked to the transplantation. We had the opportunity to observe several patients presenting with pre-epitheliomatous keratoses and cutaneous carcinomas associated with warts. We report herein on five cases that were subjected to a clinical, histological and virological study. Material and methods. Clinical and histological report. The patients were referred to use by the Kidney Transplantation Department of the Ed. Herriot Hospital (Lyon). They were examined clinically by one of us (S.E.). Virological studies. These were performed on warts, keratoses, keratoacanthomas and squamous cell carcinomas. Human papillomavirus (HPV) antigen was detected by indirect immunofluorescence using rabbit antibodies raised against group-specific HPV antigen; viral DNA was detected by in situ molecular hybridization using biotinylated probes of types 1a, 2a, 16, 18 in all cases and type 5 in 14 lesions under stringent conditions. DNA-DNA hybrids were revealed by an alkaline phosphatase enzymatic system. RESULTS: (a) Clinical data are summarized in table I (see fig. 1-5). (b) Histological examination (fig. 6-9) showed either unequivocal squamous-cell carcinoma or keratoacanthoma . The overall architecture of the lesions was reminiscent of keratoacanthoma; however the lower limit was frequently not sharply demarcated; in that area, cells contained large basophilic nuclei exhibiting atypical features and numerous mitoses. The majority of lesions had an histological appearance reminiscent of warts (table III), with upper epidermal keratinocytes being vacuolized and containing basophilic (c) The results of virological studies (fig. 10-13) are summarized in table III. HPV group specific antigen was detected merely in 5 out of 33 lesions; in contrast, in situ molecular hybridization showed that 25 out of 33 lesions contained HPV DNA, with 14 of them containing the potentially oncogenic types 16 and 18. Only 2 lesions were positive with the prove HPV 5. Discussion. The overall incidence of cancers in Kidney transplant recipients (3 p. 100) is about 100 times higher than in control populations (17). Cutaneous carcinomas account for about 50 p. 100 of cancers. This incidence increases with time after transplantation and sun exposure. The delay on onset of cutaneous malignancies is relatively long (4 to 7 years) (6,7) and becomes longer with a decreasing age of the patients at the time of transplantation, as can be noted in our cases. Apart from Blohme (1), most authors have reported a prevalence of squamous over basal-cell carcinoma. None of our patients presented basal-cell carcinoma.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2545156 TI - [What is your diagnosis? Granular cell tumor (Abrikossoff's tumor)]. PMID- 2545157 TI - [Anogenital condyloma in children and sexual abuse. Practical management]. PMID- 2545158 TI - [Diagnostic trends in the microscopic discovery of mucinous deposits]. PMID- 2545159 TI - [Sarcoma of the retroperitoneal mesenchymal soft tissues and of the cord. Apropos of 9 cases]. AB - Sarcomas of the retroperitoneal soft tissues and of the cord are rare and serious tumours. However, a better histological understanding, determination of the histological and prognostic grades and progress in chemotherapy have led to an improvement in the overall prognosis of these tumours. PMID- 2545160 TI - Spinal cord and peripheral nerve pathology in AIDS: the roles of cytomegalovirus and human immunodeficiency virus. AB - We examined spinal cords, nerve roots, or peripheral nerves of 27 patients who died with acquired immunodeficiency syndrome (AIDS) for the presence of cytomegalovirus (CMV) and human immunodeficiency virus (HIV) by immunoperoxidase techniques in paraffin-embedded tissue. Vacuolar myelopathy was seen in 8 of 26 spinal cords (31%) and microglial nodules were seen in 13 (50%). All of the patients with lateral column vacuolar myelopathy showed severe brain pathology. HIV antigens had been detected in the brains of 15 (55%) of the 27 patients but were detected in only 3 (11%) of 26 spinal cords and were not localized to regions of vacuolar myelopathy. This suggests that the vacuolar myelopathy may be due to a remote or indirect effect of HIV or other infectious agent. CMV antigens were detected in none of the patients who showed vacuolar myelopathy but were detected in 2 of the 13 with microglial nodules. Focal nerve root or peripheral nerve inflammation was seen in 7 patients; 4 had CMV antigens and none had HIV antigens. CMV appears to be an important cause of inflammatory peripheral neuropathy in AIDS patients. PMID- 2545161 TI - Characterization of ethosuximide reduction of low-threshold calcium current in thalamic neurons. AB - The mechanism by which ethosuximide reduces thalamic low-threshold calcium current (LTCC) was analyzed using voltage-clamp techniques in acutely isolated ventrobasal complex neurons from rats and guinea pigs. The ethosuximide-induced reduction of LTCC was voltage dependent: it was most pronounced at more hyperpolarized potentials and did not affect the time course of activation or inactivation of the current. Ethosuximide reduced LTCC without altering the voltage dependence of steady-state inactivation or the time course of recovery from inactivation. Dimethadione reduced LTCC by a similar mechanism, while valproic acid had no effect on LTCC. We conclude that ethosuximide reduction of LTCC in thalamic neurons is consistent with a reduction in the number of available LTCC channels or in the single LTCC channel conductance, perhaps indicating a direct channel-blocking action of this drug. Given the importance of LTCC in thalamic oscillatory behavior, a reduction in this current by ethosuximide would be a mechanism of action compatible with the known anticonvulsant effects of this drug in typical absence seizures. PMID- 2545162 TI - Cytochrome c oxidase--deficient myogenic cell lines in mitochondrial myopathy. AB - In a patient with mitochondrial myopathy, the defect of cytochrome c oxidase activity was restricted to some muscle fibers. To isolate cell lines with or without oxidase activity from a single muscle sample, primary cultured cells were transformed by replication origin-defective simian virus 40, and then cloned. The clones were examined by cytochemical staining for cytochrome c oxidase activity. Eight myogenic clones were completely devoid of activity, while the other myogenic and nonmyogenic clones were not. Deficiency of cytochrome c oxidase was stable in culture for at least a year after serial passaging. The amount of mitochondrial DNA in cytochrome c oxidase-deficient cells was the same as in control cells, and no deletion in the mitochondrial DNA was detected. Protein synthesis in mitochondria of the subunits of cytochrome c oxidase and subunit 6 of the ATP synthase complex was markedly decreased, whereas synthesis of the other subunits encoded by mitochondrial DNA was normal. These cloned cell lines provide an excellent system for clarifying the cause of mitochondrial myopathy and for investigating nuclear-mitochondrial genetic interaction. PMID- 2545163 TI - Potentiating effect of ribavirin on the anti-retrovirus activity of 3'-azido-2,6 diaminopurine-2',3'-dideoxyriboside in vitro and in vivo. AB - 3'-Azido-2,6-diaminopurine-2',3'-dideoxyriboside (AzddDAPR) is a potent and selective inhibitor of human immunodeficiency virus (HIV) replication in vitro. It also inhibits Moloney murine sarcoma virus (MSV)-induced transformation of murine C3H/3T3 embryo fibroblasts. AzddDAPR causes a marked dose-dependent suppression of MSV-induced tumor formation and mortality therewith associated in newborn mice infected with MSV. Combination of AzddDAPR with ribavirin resulted in a marked potentiation of its anti-retrovirus activity in vitro and a significant enhancement of its inhibitory effect on MSV-induced tumor formation in vivo. A slight increase in the in vivo toxicity of AzddDAPR was noted when combined with ribavirin. PMID- 2545164 TI - Antiherpes activity of chemically synthesized lipid A-subunit analogue GLA-60 in immunosuppressed mice. AB - Intraperitoneal administration of 10 micrograms GLA-60, a chemically synthesized lipid A analogue, to mice one day after treatment with 200 mg/kg of cyclophosphamide (CY) significantly increased the number of macrophages, lymphocytes and polymorphonuclear leukocytes (PMNs) in the peritoneal cavity. The intrinsic antiviral activity of macrophages against herpes simplex virus type 1 (HSV-1) as well as natural killer (NK) activity against YAC-1 target cells was stimulated by administration of GLA-60 to CY-immunosuppressed mice. When the mice were administered GLA-60 prior to HSV-1 infection, virus growth was inhibited and the mortality rate of infected mice was reduced. Thus, GLA-60 is a potent immunomodulator achieving its antiviral action through enhancement of nonspecific host defense mechanisms. Combined treatment of GLA-60 with the antiviral agent acyclovir (ACV) resulted in greater protection against HSV-1 in the CY immunosuppressed mice than did single treatment with either GLA-60 or ACV. PMID- 2545165 TI - Differential reconstitution of zidovudine-induced inhibition of mitogenic responses by interleukin-2 in peripheral blood mononuclear cells from patients with human immunodeficiency virus infection. AB - Zidovudine (ZDV), an anti-human immunodeficiency virus (HIV) therapy, has been associated with reduction in mortality and improvement of patients with acquired immunodeficiency syndrome (AIDS). The ZDV recipients, however, experience a multitude of side effects of which bone marrow suppression is the most noteworthy, especially among patients with low CD4 cell counts. The effect of ZDV and interleukin-2 (IL-2) on phytohemagglutinin (PHA)-induced proliferative response of peripheral blood mononuclear cells (PBMs) from patients with HIV infection was investigated. ZDV 0.5 micrograms inhibited 40% of PHA-induced thymidine uptake in PBMs from healthy donors or patients with HIV, irrespective of their CD4 cell counts. However, IL-2 (10 U/ml) had differential effect on PHA induced thymidine uptake that appeared to be dependent on absolute CD4 cell counts. While PBMs from patients with CD4 cell counts of 400/mm3 or more did not respond to IL-2 (low responders), IL-2 enhanced the PHA-induced thymidine uptake in PBMs from patients with CD4 cell counts less than 400/mm3 at an average of 60% (high responders). Moreover, IL-2 restored the ZDV-induced inhibition by almost 100% in the high responder group while it did not affect counts in the low responder group. The production of IL-2 in vitro, in response to PHA or recall antigens, was equivalently inhibited in both groups. These data suggest that ZDV and IL-2 could have an additive effect on immune parameters in certain groups of patients infected with HIV. The differential effect of IL-2 was independent of IL 2 receptor expression. PMID- 2545166 TI - Herpes simplex virus glycoprotein immunotherapy of recurrent genital herpes: factors influencing efficacy. AB - Recombinant herpes simplex virus type 1 (HSV-1) glycoproteins B (gB) and D (gD) were used as immunotherapeutic agents for the treatment of recurrent genital herpes in guinea pigs. Administration of a gBgD vaccine eight to 21 days after intravaginal HSV-2 inoculation significantly increased the titer of anti-HSV antibodies (P less than 0.005) while significantly reducing the frequency of subsequent herpetic recurrences (P less than 0.05). The effectiveness of gBgD immunotherapy was influenced by both the co-administration of adjuvant, the type of adjuvant, and by the timing and route of administration. These data demonstrate that recurrent HSV disease in animals with established latent infection may be favorably altered by the administration of immunogenic viral proteins. PMID- 2545168 TI - Ulcerated tumor on the scalp. Clear-cell hidradenoma. PMID- 2545167 TI - Selection of glucose-assimilating variants of Acinetobacter calcoaceticus LMD 79.41 in chemostat culture. AB - Glucose metabolism has been studied in two strains of Acinetobacter calcoaceticus. Strain LMD 82.3, was able to grow on glucose and possessed glucose dehydrogenase (EC 1.1.99.17). Glucose oxidation by whole cells was stimulated by PQQ, the prosthetic group of glucose dehydrogenase. PQQ not only increased the rate of glucose oxidation and gluconic acid production but also shortened the lag phase for growth on glucose. Strain LMD 79.41 also possessed glucose dehydrogenase but was unable to grow on glucose. Batch cultures and carbon limited chemostat cultures growing on acetate in the presence of glucose oxidized the sugar to gluconic acid, which was not further metabolized. However, after prolonged cultivation on mixtures of acetate and glucose, carbon-limited chemostat cultures suddenly acquired the capacity to utilize gluconate. This phenomenon was accompanied by the appearance of gluconate kinase and a repression of isocitrate lyase synthesis. In contrast to the starter culture, cells from chemostats which had been fully adapted to gluconate utilization, were able to utilize glucose as a sole carbon and energy source in liquid and solid media. PMID- 2545169 TI - Eruptive pruritic papules. Eruptive syringomas (eruptive hidradenomas of Jacquet and Darier). PMID- 2545170 TI - Osteoporotic bone microstructure by collagenase etching. AB - Collagenase etching has been used to show the microstructure of bone from patients suffering from primary osteoporosis. Both polished and unpolished surfaces of trabecular bone from femoral heads were treated with collagenase solution before study in the scanning electron microscope. The polished surfaces show the mineral component of this bone as small rounded units approximately 10 20 nm across, which aggregate to form a continuous phase of contiguous spheroidal particles approximately 100 nm across. Lamellations are clearly seen to be due to the removal of collagen fibres up to approximately 200 nm across, fibres in adjacent lamellae being arranged approximately perpendicular to each other. The unpolished surfaces also show small rounded units, which aggregate into rods of mineral approximately 100 nm across. Although these rods form a connected system, they are loosely packed, compatible with their being interspersed with the collagen fibres in vivo. This model for the detailed microstructure of bone is consistent with specimens from a number of other sources and shows no features unique to osteoporosis. PMID- 2545171 TI - Hydroxyapatite and urate crystal induced cytokine release by macrophages. AB - Destructive osteoarthritis is characterised by rapidly progressive joint destruction associated with intra-articular deposition of hydroxyapatite crystals. The possible role of such crystals in the pathogenesis of this condition was investigated by testing the ability of hydroxyapatite crystals to stimulate the production of bone resorbing activity from mouse peritoneal macrophages. Urate crystals were used for comparison. Culture supernatants were tested for bone resorbing activity using the mouse calvarial bone resorption assay, for interleukin 1 using a standard lymphocyte activation assay, and for prostaglandin E2 by radioimmunoassay. Culture supernatants from macrophages incubated with hydroxyapatite crystals contained dialysable bone resorbing activity, high concentrations of prostaglandin E2, but no interleukin 1 like activity. The production of the bone resorbing agent was prevented by culturing macrophages with hydroxyapatite crystals in the presence of indomethacin. By contrast, culture supernatants from macrophages incubated with urate crystals contained bone resorbing activity, which was only partly removed by dialysis, and interleukin 1 like activity. The latter was shown to be increased in culture supernatants from macrophages incubated with urate crystals in the presence of indomethacin, while production of bone resorbing activity was partially inhibited. It is considered that the bone resorbing activity liberated from macrophages stimulated by hydroxyapatite crystals can be explained by the presence of prostaglandin E2 alone, whereas the activity liberated by urate crystals is due to both prostaglandin E2 and interleukin 1. PMID- 2545172 TI - Spontaneous ruptured hepatocellular carcinoma. An appraisal of surgical treatment. AB - Spontaneous rupture with bleeding is not an infrequent complication of hepatocellular carcinoma (HCC). From May, 1972 to January, 1987, 56 symptomatic patients with ruptured HCC were managed by plication of the lesion (2 patients), ligation of either the common hepatic artery, CHAL, (39 patients), or selectively, the arterial branch supplying the tumor-bearing lobe of liver, SHAL, (8 patients), and hepatic resection, HR, (7 patients). Effective hemostasis was achieved in 68.1% of patients with the use of hepatic artery ligation (HAL). SHAL provides a comparable control of bleeding but no demonstrable reduction of postoperative organ failure when compared with CHAL. The operative treatment employed had no influence on either the postoperative rates of morbidity, mortality, or survival. However, the rate of hospital mortality was high among the four patients who had emergency anatomical lobectomy, despite the absence of severe cirrhosis. Hepatic artery ligation, either CHAL or SHAL, is a satisfactory definitive hemostatic measure for unresectable HCC when it ruptured. SHAL is probably preferred to routine emergency HR for patients with potentially resectable lesions. Nonetheless, for selected patients with easily accessible lesions, segmentectomy or subsegmentectomy could still be contemplated in the absence of severe cirrhosis. PMID- 2545173 TI - [Evolutionary changes in the structural and regulatory regions of aminoglycoside 3'-phosphotransferase type I gene of E. coli]. AB - To investigate the evolutionary relationships between the aph(3') genes from different plasmids, the nucleotide sequence of the aph(3') gene from the E. coli R plasmid was determined and compared with the known aph(3') genes of Tn903 and Tn4352. Three point mutations in the structural part of the cloned aph(3') gene caused amino acid changes in the enzyme molecule at positions 19, 27 and 48 beginning from the start codon. The structural part of the gene was followed by two stop codons and a long DNA region containing no nucleotide sequences homologous to the sequences of Tn903 or Tn4352. Both the cloned aph(3') gene and Tn4352 were limited on the left by the spacer sequence and the insertion sequence IS176. Twenty one base pairs deletion abolished the -35 sequence of the promoter suggested for the aph(3') gene of Tn4352 and resulted in formation of a fusion promoter utilizing the -35 box of IS176 and the -10 box of the aph(3') gene. The distance between the -35 and -10 sequences changed from 18 to 17 bp. Changes in the cloned aph(3') gene and the flanking DNA regions resulted in formation of a new promoter and loss of the right IS176 element. PMID- 2545174 TI - Diffuse large-cell lymphoma of B-cell origin and deficient T-cell function in a patient with rheumatoid arthritis. AB - An increased incidence of non-Hodgkin's lymphoma has been described in patients with rheumatoid arthritis. Mechanisms related to abnormal immune regulation have been postulated, but no patients with rheumatoid arthritis and lymphoma have been previously well characterized immunologically. We describe here a patient with long-standing rheumatoid arthritis in whom a B-cell diffuse large-cell lymphoma developed. He was found to have a severe T-cell immunodeficiency and evidence of persistent Epstein-Barr virus infection. Epstein-Barr nuclear antigen was not found to be present within lymphoma cells. The combination of defective T-cell function and persistent Epstein-Barr virus infection may have predisposed this patient with rheumatoid arthritis to the development of a malignant clone of B lymphocytes. PMID- 2545175 TI - A primary respiratory Na+ pump of an anaerobic bacterium: the Na+-dependent NADH:quinone oxidoreductase of Klebsiella pneumoniae. AB - Membranes of Klebsiella pneumoniae, grown anaerobically on citrate, contain a NADH oxidase activity that is activated specifically by Na+ or Li+ ions and effectively inhibited by 2-heptyl-4-hydroxyquinoline-N-oxide (HQNO). Cytochromes b and d were present in the membranes, and the steady state reduction level of cytochrome b increased on NaCl addition. Inverted bacterial membrane vesicles accumulated Na+ ions upon NADH oxidation. Na+ uptake was completely inhibited by monensin and by HQNO and slightly stimulated by carbonylcyanide-p trifluoromethoxy phenylhydrazone (FCCP), thus indicating the operation of a primary Na+ pump. A Triton extract of the bacterial membranes did not catalyze NADH oxidation by O2, but by ferricyanide or menadione in a Na+-independent manner. The Na+-dependent NADH oxidation by O2 was restored by adding ubiquinone 1 in micromolar concentrations. After inhibition of the terminal oxidase with KCN, ubiquinol was formed from ubiquinone-1 and NADH. The reaction was stimulated about 6-fold by 10 mM NaCl and was severely inhibited by low amounts of HQNO. Superoxide radicals were formed during electron transfer from NADH to ubiquinone 1. These radicals disappeared by adding NaCl, but not with NaCl and HQNO. It is suggested that the superoxide radicals arise from semiquinone radicals which are formed by one electron reduction of quinone in a Na+-independent reaction sequence and then dismutase in a Na+ and HQNO sensitive reaction to quinone and quinol. The mechanism of the respiratory Na+ pump of K. pneumoniae appears to be quite similar to that of Vibrio alginolyticus. PMID- 2545176 TI - Nephroblastomatosis. An overview. PMID- 2545177 TI - Cushing's syndrome. Diagnosis of the atypical patient. PMID- 2545178 TI - Bilateral diffuse nephroblastomatosis, pancortical type. A case report with immunohistochemical investigations. AB - A variation of Perlman's syndrome of the pancortical type is reported in a male neonate whose parents were cousins. The patient was the product of a 35-week pregnancy, the Apgar score was 3, and the patient died of respiratory failure one hour and 12 minutes after delivery. Autopsy revealed bilateral diffuse nephroblastomatosis, pancortical type, associated with malformations (usually facial), congenital anomalies of the heart, hepatosplenomegaly, pancreatic islet cell hyperplasia, bilateral cryptorchidism, and hyperflexibility of the left knee joint. Renal immunohistochemical investigations revealed positive bindings with peanut and soybean agglutinins and epithelial membrane antigen along the luminal surface of the epithelium in the moderately differentiated tubules, but not in blastoma or primitive epithelium. PMID- 2545179 TI - Spurious elevation of plasma immunoreactive adrenocorticotropic hormone in cyclic Cushing's syndrome. AB - We studied a 40-year-old woman with cyclic Cushing's syndrome who demonstrated abnormal high-dose dexamethasone suppression and metyrapone stimulation tests. These results, associated with persistent elevations of plasma adrenocorticotropic hormone (ACTH) levels, suggested ectopic secretion of ACTH. Surprisingly, an adrenal adenoma with atrophy of the contralateral adrenal gland was found at exploratory laparotomy. Subsequent ACTH determinations that extract ACTH from the plasma before assay suggested that the apparent increase in ACTH concentration in our routine assay was due to the presence of an interfering substance(s). We conclude that the diagnosis of Cushing's syndrome continues to depend on a battery of adrenal function tests and radiographic procedures and recommend that measurements of ACTH be performed only after extraction of ACTH from specimens. PMID- 2545180 TI - Oncological aspects of immediate breast reconstruction following mastectomy for malignancy. AB - One hundred eighteen women treated with mastectomy and immediate breast reconstruction for carcinoma were evaluated for recurrence of disease and survival. Fourteen women (12%) suffered relapse of their cancer and 10 patients (9%) died of their disease during a median follow-up of 2.3 years. All seven local recurrences (6%) were detected at an early stage and treated without removal of the prosthesis. Recurrence of disease occurred more frequently in patients with involved axillary lymph nodes and larger tumors. Patient survival was adversely affected by nodal metastasis and the absence of tumor estrogen receptors. Adverse outcome in this series correlated to known prognostic factors for breast cancer. Disease-free and overall survivals were comparable with our previous experience with mastectomy alone for breast carcinoma. In the absence of any apparent negative impact on patient outcome, and because of the well documented positive psychosocial benefit of immediate reconstruction, this procedure should be routinely offered to women with operable breast cancer. PMID- 2545181 TI - Effect of lead exposure on some biological indices related to porphyrin metabolism and the activity of erythrocyte pyrimidine 5'-nucleotidase in the mice. AB - The urinary excretion of delta-aminolevulinic acid (ALA) and coproporphyrin (CP) isomers (I and III) was investigated in mice exposed to lead in drinking water (200 and 500 ppm) for 14 or 30 days. Furthermore, the inhibition of pyrimidine 5' nucleotidase (P5N) and delta-aminolevulinic acid dehydratase (ALAD) was studied in the same mice. In the lead-exposed mice, urinary excretion of ALA was significantly elevated (12-times control), while that of CP remained unchanged. This discrepancy on the urinary excretion may be of interest. The P5N activity in both erythrocytes and bone marrow cells was significantly inhibited by exposure to lead. However, the degree of inhibition was greater in the erythrocytes (45% inhibition) than in the bone marrow cells (25% inhibition). The inhibition of ALAD was much higher in erythrocytes (90% inhibition) than in liver (20-40% inhibition). The increase in erythrocyte protoporphyrin and the decrease in both hemoglobin and hematocrit were not observed in mice exposed to lead under the above conditions. PMID- 2545183 TI - [Relation of steroid receptors, tumor ploidy, and cell proliferation in various histologic types of breast cancer]. AB - The receptor status of the tumors examined in 74 patients with breast cancer was found to correlate with its ploidy and proliferative activity as evidenced by flow cytometry and it failed to correlate with a histological tumor type and metastatic involvement of regional lymph nodes. In terms of DNA content, the receptor-positive tumors were more frequently diploid, the receptor-negative ones were aneuploid. The proliferation index (PI) for the receptor-negative tumors was significantly higher than that for the receptor-positive ones. The diploid tumors showed lower proliferative activity than the aneuploid ones. Among the aneuploid tumors, PI for infiltrative lobular cancer was significantly lower than that for infiltrative duct cancer. Early in invasion, duct carcinoma was predominantly diploid, of grade I-IIA malignancy and more infrequently metastasized into regional lymph nodes. Grade IIB-III malignant tumors were chiefly aneuploid and more frequently metastasized into regional lymph nodes. PMID- 2545184 TI - [Ultrastructural characteristics of virus-producing lymphocytes in bovine leukemia]. AB - The ultrastructure of peripheral lymphocytes and lymph was examined in 35 head of cattle in health and chronic lymphoid leukemia. In the latter, ultrastructural alterations were detected in the nuclei and cytoplasmic organelles. The alterations appeared as Golgi apparatus hypertrophy, mitochondrial polymorphism, myelinoid formations. There were nuclear pockets, invaginations in the nuclei of leukemic lymphocytes. A correlation was established between the number of cells having nuclear pockets and the virus production of lymphoid cells in chronic lymphoid leukemia. There is evidence that the existence of nuclear pockets along with other indicators may be useful in assessing the development of leukemias in cattle. The morphogenesis of bovine leukemia virus is presented. PMID- 2545182 TI - Modulation of gentamicin nephrotoxicity by chronic inhibition of angiotensin-I converting enzyme in rat. AB - Perindopril, a new specific and potent inhibitor of angiotensin-I-converting enzyme, was used to evaluate the possible participation of inhibition of the renin-angiotensin system in the development of aminoglycoside-induced renal failure. Kidney function, morphology and biochemistry were evaluated at regular intervals throughout the study. Perindopril was given orally to rats at a daily dose of 2 mg/kg for 15 days prior to and during 15-day gentamicin treatment given intraperitoneally at a daily dose of 50 mg/kg. Perindopril treatment alone induced no modification in renal function or structure. Gentamicin treatment alone induced typical renal lesions which were scored as moderate and a slight but significant decrease in ACE blood levels. Concurrent treatment with perindopril and gentamicin induced a greater drop in ACE blood levels than after the administration of perindopril alone and produced more marked renal impairment than after the administration of gentamicin alone. These observations suggest that the integrity of the renin-angiotensin system may play an important role in limiting kidney injury during aminoglycoside-induced nephrotoxicity. PMID- 2545185 TI - [Status of the hypophyseo-adrenal system and functional morphology of the adrenal glands in residents of Northern European areas of the USSR]. AB - Serum corticotropin and cortisol levels were measured in 20-40-year old healthy residents (144 males and 361 females) in the North. A functional Synacthen Depot test was performed in 5 volunteers. The adrenal glands from 56 males and 8 females of the same age who had died from accidents were pathohistologically and stereometrically examined. The signs of functional tension were established in the pituitary-adrenal cortex system. The Synacthen Depot test determined sufficient potentials of the organ to show responses to specific stimulation. Morphologically, a number of changes were found, which might be regarded as a prerequisite to the development of functional and pathological disturbances in the above-mentioned link of the endocrine system. PMID- 2545187 TI - Achieving health by hope. PMID- 2545186 TI - Metamorphopsia and palinopsia. Association with periodic lateralized epileptiform discharges in a patient with malignant astrocytoma. AB - A patient with a right parietal glioma developed palinopsia and various forms of metamorphopsia that were temporally related to the electrographic presence of periodic lateralized epileptiform discharges. These symptoms occurred in an alert woman with normal visual fields. The literature shows that epileptic events can produce these symptoms. We believe that continuous repetitive electrical stimulation caused our patient's unusual symptoms by altering the association pathways through which visual data are experienced. To our knowledge, the association between periodic lateralized epileptiform discharges and metamorphopsia or palinopsia has heretofore not been reported. PMID- 2545188 TI - Mutations, epimutations, and the developmental programming of the maize Suppressor-mutator transposable element. AB - Information about the structure, function and regulation of the maize Suppressor mutator (Spm) transposable element has emerged from the genetic and molecular characterization of both deletion mutations and an unconventional type of reversible genetic change (epimutation). The element is subject to an epigenetic mechanism that can either stably inactivate it or specify one of a variety of heritable programs of differential element expression in development. The essay explores the relationship between the Spm element's epigenetic developmental programming mechanism and the determinative events central to plant development and differentiation. PMID- 2545189 TI - Regulation of the Ca2+ pump ATPase by cAMP-dependent phosphorylation of phospholamban. AB - Ca2+ transients in myocardial cells are modulated by cyclic AMP-dependent phosphorylation of a protein in the sarcoplasmic reticulum. This protein, termed phospholamban, serves to regulate the Ca2+ pump ATPase of this membrane, thus altering the mode of Ca2+ transients and the myocardial contractile response. Elucidating the structure of phospholamban and its intimate interaction with the Ca2+ pump ATPase should provide the basis for understanding, at the molecular level, how the cAMP system contributes to excitation-contraction coupling in muscle cells. PMID- 2545190 TI - Bromoconduritol treatment delays intracellular transport of secretory glycoproteins in human hepatoma cell cultures. AB - Previous studies in our laboratory have shown that specific glycan structures are required for the normal secretion of some glycoproteins. Bromoconduritol is known to inhibit the removal of the innermost glucose moiety from the Glc3Man9(GlcNAc)2 precursor of N-linked glycoproteins. We have used this inhibitor to investigate the possible role of glycan structure in the intracellular transport of secretory glycoproteins of Hep G2 cultures. Cells were pretreated with 1mM bromoconduritol for 1h, pulsed with [35S]-methionine for 10min and chased for varying intervals. Specific glycoproteins and albumin were immunoprecipitated from the cell lysate and medium. We found that bromoconduritol-treatment inhibited the secretion of alpha 1-protease inhibitor, ceruloplasmin, alpha 2-macroglobulin, transferrin, and alpha-fetoprotein. Apparently, the glucosylated high-mannose intermediate is not secreted, since glycoproteins in the medium are of complex form. We conclude that the removal of the innermost glucose residue from secretory glycoprotein represents an important regulatory step in the intracellular transport pathway. PMID- 2545191 TI - Effect of interferon inducers on superoxide anion generation from rat liver microsomes detected by lucigenin chemiluminescence. AB - Microsomal superoxide anion (O2-) production was detected using the chemiluminigenic probe, bis-N-Methylacridinium nitrate (lucigenin). Superoxide dismutase (SOD) inhibited 55% of the light emission but in the presence of a detergent (Triton X100) SOD inhibited the light emission by 94%. Lucigenin chemiluminescence from rat liver microsomes supplemented with NADPH was found to be selective and sensitive in detecting the O2- production. Treatment of rats with poly IC and LPS resulted in a decrease of the hepatic microsomal cytochrome P450 content by 44% and 37% respectively. The decrease in the cytochrome P450 contents was accompanied by a decrease in LgCl from the hepatic microsomal fractions by 61% for the poly IC and by 51% for the LPS treated rats. This is the first report to demonstrate that decreased P450 in the presence of normal amounts of cytochrome P450(c) reductase produce correspondingly less O2- from the microsomes. PMID- 2545192 TI - Endothelin-1 induces vasoconstriction through two functionally distinct pathways in porcine coronary artery: contribution of phosphoinositide turnover. AB - Endothelin-1 (ET1)-induced contraction of isolated porcine coronary artery strips was previously reported to be mainly dependent on extracellular Ca2+. However, even in a Ca2+-free, EGTA-containing solution relatively high concentrations of ET1 induced a weak vasoconstriction, which was markedly but not completely inhibited by pretreatment with caffeine. Over similar dose ranges, ET1 stimulated the production of inositol phosphates in a dose-dependent manner in intact arterial tissues, which was independent of extracellular Ca2+ and was not affected by receptor blockers such as atropine, methysergide and diphenhydramine. Moreover, ET1 was shown to induce an increase in 1,2-diacylglycerol. These results indicate that the activation of ET1 receptors on porcine coronary artery smooth muscle causes phosphoinositide breakdown, leading to intracellular Ca2+ mobilization and protein kinase C activation. It is suggested that phospholipase C-mediated phosphoinositide breakdown as well as previously reported activation of voltage-dependent Ca2+ channels are involved in the mechanism of ET1-induced vasoconstriction. PMID- 2545193 TI - Photoaffinity labeling of platelet activating factor binding sites in rabbit platelet membranes. AB - A photoreactive, radioiodinated derivative of platelet activating factor (PAF), 1 O-(4-azido-2-hydroxy-3-iodobenzamido)undecyl-2-O-acetyl-sn- glycero-3 phosphocholine ([125I]AAGP), was synthesized and used as a photoaffinity probe to study the PAF binding sites in rabbit platelet membranes. The nonradioactive analog, IAAGP, induced rabbit platelet aggregation with an EC50 value of 3.2 +/- 1.9 nM as compared to 0.40 +/- 0.25 nM for PAF. Specific binding of [125I]AAGP to rabbit platelet membranes was saturable with a dissociation constant (Kd) of 2.4 +/- 0.7 nM and a receptor density (Bmax) of 1.1 +/- 0.2 pmol/mg protein. Photoaffinity labeling of platelet membranes with [125I]AAGP revealed several 125I-labeled components by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A protein species with apparent molecular weight of 52,000 was consistently observed and inhibited significantly by unlabeled PAF at nanomolar concentrations. The labeling was specific since the PAF antagonists, SRI-63,675 and L-652,731, at 1 uM also blocked the appearance of this band; whereas lysoPAF was not effective at the same concentration. These results suggest that the binding sites of PAF receptor in rabbit platelets reside in the polypeptide of Mr = 52,000. PMID- 2545194 TI - Hydroxyl radical and leukotoxin biosynthesis in neutrophil plasma membrane. AB - We measured O2-, H2O2, .OH and leukotoxin biosynthesis in neutrophil plasma membrane. O2- was produced by respiratory burst oxidase in the membrane coupling with NADPH oxidation. Leukotoxin was biosynthesized in the system containing linoleate. Addition of SOD into the system doubled the amount of leukotoxin synthesized. Further addition of catalase into the system decreased leukotoxin formation. .OH and leukotoxin formation in the system was substantially increased by the presence of cytochrome c. In addition, leukotoxin was detected in the non biological reaction mixture containing H2O2 and linoleate in the presence of heme iron. In this mixture, .OH and leukotoxin formation also showed a good correlation. From these results, it is evident that, in neutrophil cell membrane, leukotoxin is synthesized by .OH with linoleate. PMID- 2545195 TI - Photoaffinity labelling of gonadotropin releasing hormone binding sites in human epithelial ovarian carcinomata. AB - A photoaffinity labelled derivative of [D-Lys6]-GnRH was prepared with a bifunctional photolabile reagent (4-azidobenzoyl)-N-hydroxysuccinimide. In rat pituitary membranes, this analog retained high binding affinity (Ka = 0.12 x 10(9) M-1) consistent with a single class of receptors. The analog was iodinated and used for the identification of GnRH binding sites in human epithelial ovarian carcinomata. By sodium dodecyl sulfate electrophoresis in 10% polyacrylamide gel the presence of two labelled components could be demonstrated: a high molecular weight component of 63,200 and a smaller component of 46,000. Competition experiments with unlabelled ligand suggest that it is the high molecular weight component which specifically binds GnRH. PMID- 2545196 TI - Na+-H+ antiport during fertilization of the sea urchin egg is blocked by W-7 but is insensitive to K252a and H-7. AB - Fertilization of the sea urchin egg initiates or accelerates a number of metabolic activities, which have been causally linked to a rise in cytoplasmic pH due to increased Na+-H+ antiport. Two possible regulatory pathways linking sperm egg fusion to the activity of the antiporter are activation of protein kinase C (PKC) and Ca2+, calmodulin (CaM)-dependent kinase. This report presents the effects of protein kinase inhibitors on antiporter activation during fertilization and treatment with PKC agonists, dioctanoylglycerol or phorbol diester. Protein kinase inhibitors, K252a and H-7 blocked the action of PKC agonists, without inhibiting cytoplasmic alkalinization during fertilization. In contrast, W-7 blocked fertilization-induced rise in cytoplasmic pH, without altering the actions of PKC agonists. These results suggest that the Na+-H+ antiporter may be regulated by PKC or Ca2+, CaM-dependent kinase activities, but activation of the antiporter during fertilization is Ca2+, CaM-dependent, despite production of diacylglycerols by hydrolysis of phosphatidylinositols. PMID- 2545197 TI - Inhibition of fibrinogen binding to human platelets by blockage of Na+/H+ exchange. AB - Binding of ADP to platelets enhances the binding of fibrinogen to Gp IIb-IIIa, the specific platelet receptor for adhesive proteins. The linkage between ADP and fibrinogen binding is indirect since ADP does not bind to the same receptor as fibrinogen. We have recently proposed that a third component, once affected by ADP binding, induces a conformational transition of the fibrinogen receptor from the low to the high affinity state, which is responsible for platelet aggregation [De Cristofaro, R., Landolfi, R., Castagnola, M., De Candia, E., Di Cera, E., & Wyman, J. (1988) Proc. Natl. Acad. Sci. USA 85, 8473-8476]. In the present study we provide evidence that this component should be identified with the platelet Na+/H+ antiport. Inhibition of the antiport by pharmacological agents such as amiloride, or else by decreasing extracellular Na+, results in a marked decrease of fibrinogen binding to platelets. PMID- 2545198 TI - Two distinct types of endothelin receptors are present on chick cardiac membranes. AB - Competitive displacement experiments of 125I-endothelin (ET)-1, -2, or -3 binding to chick cardiac membranes were performed with unlabeled ET-1, -2, -3, and sarafotoxin S6b (STX) as competitors. 125I-ET-1 and -2 binding was competitively inhibited by increasing concentrations of these unlabeled peptides in the same order; i.e. ET-2 greater than or equal to ET-1 greater than ET-3 greater than STX. In contrast, the order of potency in displacing 125I-ET-3 binding was ET-3 greater than ET-2 greater than or equal to ET-1 greater than STX. Affinity labeling of the membranes by cross-linking with 125I-ET-1 and -2 via disuccinimidyl tartarate yielded one major specific band with an apparent Mr = 53,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by autoradiography. On the other hand, affinity labeling with 125I-ET-3 showed that two major and one minor bands of Mr = 34,000, 46,000, and 53,000, respectively, were specifically labeled. These results indicate the presence of two distinct types of ET receptors, one of which has higher affinity for ET-1 and -2 than ET-3 and the other is conversely ET-3-preferring. PMID- 2545199 TI - Identification of the two types of specific receptor for activin/EDF expressed on Friend leukemia and embryonal carcinoma cells. AB - Activin and inhibin are polypeptide factors which control the release of follicle stimulating hormone(FSH) from pituitary cells. The recent finding that erythroid differentiation factor(EDF) is identical to activin showed the multifunctional feature of this protein. To identify the specific receptor for activin, the binding of 125I-labeled activinA was investigated for a number of culture cell lines. Friends leukemia cell, which can be differentiated by activin, and embryonal carcinoma(EC) cells(PCC3, P19 and F9), were found to express 3500 20,000 per cell of activin receptors. Scatchard plot analysis of the binding data shows that the receptors on those cells could be divided into two groups with different Kd values. The Kd values of high and low affinity receptors are 0.15 0.4 nM and 1.5-3.0 nM respectively. The proportion of the number of the high and low affinity receptors was varied in each cell. Inhibin was able to compete for activin binding to both types of receptors, although the binding affinity was about 50-200 fold lower than that of activinA. Transforming growth factor-beta had no binding ability to the activin receptors. PMID- 2545200 TI - Effect of hexadecane on piglet epidermal ornithine decarboxylase activity. AB - Topical application of hexadecane has been shown to induce hyperproliferation and hyperkeratosis in rodent skin. The application of hexadecane to epidermis from the backs of piglets less than 1 week old resulted in a rapid biphasic-rise in the level of ornithine decarboxylase (ODC) activity. The second phase of the elevation of activity was suppressed by cycloheximide indicating that it resulted from de novo protein synthesis. The first, cycloheximide-insensitive phase presumably represents activation of existing enzyme. The activation of this latent ODC by hexadecane was independent of extracellular calcium. A similar degree of activation was observed using the bivalent-cation ionophore A23187 which augmented the hexadecane effect implicating a rise in intracellular calcium concentration as a possible cause for the activation possibly via the receptor mediated phospholipid hydrolysis. The time-course of the ODC activation also corresponded with a rapid fall in cAMP levels indicating a possible role for cAMP in ODC regulation. PMID- 2545201 TI - First pharmacological characterization of TRH receptors linked to phosphoinositide hydrolysis in GH3 pituitary cells using agonist specificity of eight TRH analogs. AB - The agonist/antagonist properties of TRH and 8 TRH analogs were ascertained in GH3 cells using accumulation of [3H]inositol phosphates ([3H]IPs) as an index of receptor activation. All TRH analogs, except diketopiperazine (DKP), were full agonists producing similar maximum stimulation (6.5 +/- 1.1-fold) of [3H]IP production. Concentrations of peptides producing half-maximal stimulation of phosphoinositide (PI) hydrolysis were (nM; means +/- SEM): MeTRH (2.4 +/- 0.4); MK-771 (7.3 +/- 0.6); TRH (26.6 +/- 9.2); RX77368 (90.2 +/- 13.9); CG3703 (274.5 +/- 104.4); N-Val2-TRH (2400 +/- 870); CG3509 (16500 +/- 3400); TRH free acid (17.3, 11.0 microM), DKP (greater than 1 mM). The rank order of potency of TRH analogs at inducing PI turnover was similar to that for competition of [3H]MeTRH binding to brain and pituitary homogenates reported previously, thus indicating the identification of functional TRH receptors. These data suggest that while the modifications of the C- and N-termini of the TRH molecule (resulting in MK-771, RX77368, CG3703, CG3509) reduce the apparent affinities of these compounds, the latter still retain considerable agonist activity, and in the case of MK-771 remain equipotent or become slightly more potent than TRH. This study, therefore, constitutes the first to demonstrate the biological activity of the 8 peptide analogs of TRH using the PI turnover technique in cultured clonal cells. PMID- 2545202 TI - Existence of a polar digitalis-like factor in mammalian hypothalamus. AB - We were able to partially purify a polar digitalis-like factor from rat and bovine hypothalami based on the capacity to inhibit [3H]ouabain binding to intact human erythrocytes. This factor was characterized in reference to the digitalis like factor that we have isolated and reported on. Hypothalamic factor shared digitalis-like activities and physicochemical properties with the one derived from human urine and mammalian plasma. These findings strongly suggest that a polar digitalis-like factor identical to the circulatory factor does exist in mammalian hypothalamus. PMID- 2545203 TI - Conformation studies on bombesin receptor antagonists: 500 MHz NMR and CD characterization of synthetic (D-Phe12, Leu14)-bombesin. AB - The conformation flexibility of the tetradecapeptide hormone bombesin and its synthetic antagonist (DPhe12, Leu14)-bombesin has been studied using nuclear magnetic resonance and circular dichroism techniques. The spectral features observed indicate that the ordered structure present in the C-terminal pentapeptide moiety of native BBS is lost in the (DPhe12, Leu14) analog. PMID- 2545204 TI - Sodium and potassium regulation of guanine nucleotide-stimulated adenylate cyclase in brain. AB - The present study examines the influence of potassium and sodium ions on guanine nucleotide regulation of adenylate cyclase in various brain regions, including the locus coeruleus (LC), dorsal raphe (DR), ventral tegmentum (VT), hippocampus (HP), frontal cortex (FC), substantia nigra (SN), neostriatum (NS) and cerebellum (CB). Guanine nucleotide regulation of adenylate cyclase was highest in the LC, DR and VT and lowest in NS and CB. Sodium and potassium ions were found to stimulate basal or GTP-activated adenylate cyclase in NS and SN, whereas the cations were found to specifically inhibit guanine nucleotide-stimulated enzyme activity in all other brain regions with the exception of CB, where there was no effect. With regard to stimulation of adenylate cyclase, lithium was more potent than sodium which was more potent than potassium in SN and NS. With regard to inhibition of the enzyme, potassium was equipotent to lithium which was greater than sodium in the other brain regions examined. Both stimulatory and inhibitory effects of cations in the different regions were significant (P less than 0.05) at 30 mM and were maximal at 90-120 mM. Sodium ion inhibition of GTP-stimulated adenylate cyclase in LC and DR was partially blocked by pertussis toxin treatment, whereas cation stimulation in NS was not affected by the toxin. The results demonstrate marked region-specific effects of sodium and potassium on adenylate cyclase, which could occur at either G-proteins or the catalytic unit of the enzyme. The possibility that ion fluxes alter G-protein function is discussed. PMID- 2545205 TI - Guanine nucleotide-mediated inhibition of opioid agonist binding. Modulatory effects of ions and of receptor occupancy. AB - We have analysed the potency of GTP, GDP and their analogues in reducing [3H]DADLE binding to opioid receptors in NG 108-15 cell membranes. Under conditions where non-specific hydrolysis and transphosphorylation is inhibited, the following rank order of potency was found: GDP greater than or equal to GTP gamma S greater than GTP greater than GDP beta S greater than or equal to GDPNH2 greater than GppNHp much greater than GMP. Remarkably, the slopes for the inhibition curves of GTP, GDP and their thiosubstituted analogues, but not of GDPNH2 and GppNHp, were extremely shallow, indicating either negative cooperativity or the existence of two states for the guanine nucleotide binding proteins, that both can mediate the effect of nucleotides on agonist receptor binding. The potencies of the different guanine nucleotide analogues, except that of GppNHp, were increased by the presence of sodium or chloride ions in the assay medium. Magnesium also affected GTP-mediated inhibition of opioid agonist binding since it decreased the IC50 of the nucleotide and steepened the slope of the inhibition curve. The IC50s of nucleotides and the slopes of their inhibition curves were also dependent on the extent of receptor occupancy by the agonist. From these data we conclude that (1) either diphospho- or triphosphonucleotides can regulate agonist binding. (2) Magnesium, sodium and chloride, by acting at different components of the receptor/G protein complex produce similar effects on nucleotide mediated regulation of agonist binding. (3) A mutual influence exists between receptor occupancy by agonists and G protein-mediated guanine nucleotide effect on the receptor. PMID- 2545206 TI - Mechanistic aspects of the synergistic antiviral effect of xanthates and monocarbonic acids. AB - The xanthate tricyclodecan-9-yl-xanthogenate (D609) displays antiviral and antitumoral properties that are inversely proportional in vitro to the serum concentration. Accordingly, it has been found that D609 binds to serum albumin. Recently, we have reported that D609, in combination with undecanoic acid, has a synergistic antiviral effect, which appears, as shown here, to be due to competition for the same binding domain on serum albumin. Furthermore, undecanoic acid fosters the binding of D609 to the cell. Both the competition of D609 with monocarbonic acid for binding on serum albumin and the enhanced binding of xanthate to the cell are dependent, in accordance with previously reported results, on the chain length of the fatty acids. Eleven to 14 C-atoms (undecanoic, lauric and myristic acid) were found to be appropriate while shorter (C6) and larger (C18) monocarbonic acids were shown to lack synergistic properties. PMID- 2545207 TI - (E)-5-(2-bromovinyl)uridine requires phosphorylation by the herpes simplex virus (type 1)-induced thymidine kinase to express its antiviral activity. AB - (E)-5-(2-Bromovinyl)uridine (BVUrd), the riboside counterpart of (E)-5-(2 bromovinyl)-2'-deoxyuridine (BVdUrd), effected a dose-dependent inhibition of viral progeny formation and viral DNA synthesis in herpes simplex virus type 1 (HSV-1, strain KOS)-infected human (E6SM) diploid fibroblast cells. BVUrd was directly phosphorylated in HSV-1-infected cells, presumably by the virus-encoded thymidine kinase (TK), since (i) BVUrd was not phosphorylated by extracts of cells infected with a HSV-1 strain deficient in TK expression and (ii) the phosphorylation was inhibited by a polyclonal anti-HSV-1 antibody. Within the HSV 1-infected cells, BVUrd was incorporated into the viral DNA as BVdUMP (BVdUrd 5' monophosphate). This incorporation may account for the antiviral action of BVUrd, and implies that, following its initial phosphorylation by the viral TK, BVUrd is converted to its 2'-deoxy counterpart, most likely at the 5'-diphosphate level (BVUDP----BVdUDP). PMID- 2545208 TI - Alteration of cyclic GMP metabolism by CD-349, a novel calcium antagonist, and by sodium nitroprusside in bovine intrapulmonary artery and vein. AB - The effects of 2-nitratopropyl 3-nitratopropyl 2,6-dimethyl-4-(3-nitrophenyl)-1,4 dihydropyridine-3,5-dicarboxylate (CD-349) and sodium nitroprusside (NP) on cyclic GMP (cGMP) metabolism in bovine intrapulmonary artery (BPA) and vein (BPV) were examined. CD-349 inhibited cGMP phosphodiesterase (PDE) activity in BPA and BPV. In the latter, about 40% of the cGMP PDE activity was Ca2+ dependent. The inhibition of cGMP PDE activity by CD-349 also depended on Ca2+. The inhibitory effect of CD-349 was more potent than that of nicardipine or nifedipine. The conversion of cGMP from GTP in the homogenates of BPA and BPV was stimulated by NP in a concentration-dependent manner. The NP-induced cGMP formation was stimulated further by CD-349. This effect of CD-349 depended on Ca2+ in the BPV but not in the BPA. The NP-induced elevation of cGMP levels in the tissue preparations of BPA and BPV was also potentiated by CD-349. These results suggest that CD-349 inhibited Ca2+-dependent cGMP PDE activity and that the levels of cGMP were elevated in vascular smooth muscle, particularly when guanylate cyclase was activated. PMID- 2545209 TI - Phosphorylation of arabinosyl guanine by a mitochondrial enzyme of bovine liver. AB - Extracts of mitochondria isolated from bovine liver were shown to phosphorylate araG, forming araGMP as the sole product. When other nucleosides were used as competitors with araG as the substrate for phosphorylation, deoxycytidine, deoxythymidine and guanosine were not significantly inhibitory. However, the phosphorylation of araG was blocked by deoxyguanosine, deoxyadenosine and deoxyinosine. Deoxyguanosine was shown to be a competitive inhibitor of araG phosphorylation (apparent Ki for deoxyguanosine = 9 microM; apparent Km for araG = 66 microM). Likewise, araG was determined to be a competitive inhibitor of mitochondrial deoxyguanosine kinase activity (apparent Km for deoxyguanosine = 16 microM; apparent Ki for araG = 55 microM). These data suggest that the two nucleosides were phosphorylated by the same enzyme. Disc gel electrophoresis showed that the phosphorylating activity for araG migrated with deoxyguanosine kinase activity. The pH profiles of the araG and deoxyguanosine kinase activities were dissimilar. The optimum pH for deoxyguanosine kinase was 5.5; for araG kinase, it was 8.0. Collectively, these data suggest that araG is phosphorylated by mitochondrial deoxyguanosine kinase; however, separate forms of the enzyme or different reaction conditions may be involved in the optimal activities of the two catalytic events. PMID- 2545210 TI - Effect of cryptosin on Na+, K+-ATPase--a 31P NMR spectroscopic study. PMID- 2545211 TI - A reply: "Do quinolones bind to DNA?"--Yes. PMID- 2545212 TI - Effect of the acetylcholinesterase inhibitor, soman, on plasma levels of beta endorphin and adrenocorticotropic hormone (ACTH). PMID- 2545213 TI - [Immobilized oligonucleotides as affinity sorbents for restriction endonucleases]. AB - Affinity chromatography of IIS type restriction endonucleases is proposed. It is shown that endonucleases HgaI, FokI, and SfaNI have affinity to the matrix with immobilized oligonucleotides which contain the endonuclease's recognition sites resistant to the hydrolysis. PMID- 2545214 TI - [Primary structure of the A22 RNA polymerase gene of the foot and mouth disease virus]. AB - Complete nucleotide sequence of gene RNA polymerase for the foot-and-mouth disease virus subtype A22 has been determined. PMID- 2545215 TI - Infantile cytochrome c oxidase deficiency with neonatal death. AB - A newborn male presented with severe respiratory insufficiency, generalized muscle weakness, and lactic acidemia. Immediately after admission, he was placed on a respirator because of respiratory arrest. He deteriorated rapidly and died 75 hours after birth. There was notable variation in fiber size and an increased number of type 2C fibers in the quadriceps femoris muscle obtained at autopsy; however, no ragged-red fibers were observed with modified Gomori trichrome staining. Markedly decreased cytochrome c oxidase activity was demonstrated in skeletal muscle by biochemical and histochemical studies, while cardiac muscle demonstrated normal cytochrome c oxidase activity. Mitochondrial myopathy should be considered in the differential diagnosis of patients with neonatal respiratory distress syndrome. PMID- 2545216 TI - [Synovial sarcomas of the neck]. AB - The synovial sarcoma is a mesenchymal tumour that is very rare in the head and neck region. This is a report on two cases. In a 30-year old patient the tumour arose near the thyroid gland and the hypopharynx. He died in spite of surgical removal, radiotherapy and chemotherapy, one year after the first symptoms. An 8 year old boy was affected by a synovial sarcoma near the hyoid bone and is without recurrence after surgical removal, radiotherapy and chemotherapy. Subsequently, the main diagnostic characteristics of these tumours are discussed. A better prognosis of the tumours associated with the hyoid bone is a remarkable fact. PMID- 2545217 TI - Residual motion of hemoglobin-bound spin labels as a probe for protein dynamics. AB - The residual motion of spin labels bound to cysteine beta 93 of methemoglobin and oxyhemoglobin has been analyzed as a function of temperature and hydration. The rotational diffusion of the whole protein molecule has been prevented or restricted by crystallization, lyophilization or by high viscosity of the solution. The residual motion of the labels is characterized by an angle of the limited motion cone and their rotational correlation time using computer simulations of the EPR spectra. Two types of motion can be separated due to different correlation times and different dependences on temperature and hydration. One of these motional mechanisms can be shown to be determined by protein fluctuations. Correlation times of these fluctuations decrease from 2 X 10(-8) s at T = 220 K to 10(-9) s at T = 300 K in the samples of high water concentration. Strong correlation between the properties of the hydration shell and these fluctuations are observed. PMID- 2545218 TI - The significance of DNA measurements in a histologically defined subset of infiltrating ductal carcinomas of the breast with long-term follow-up. AB - The nuclear DNA content and other karyometric parameters were evaluated in a histologically homogeneous group of invasive ductal carcinomas of the breast from 13 patients who survived 25 years after radical mastectomy and from 13 controls matched for histologic tumor grade, lymph node status, tumor size and patient age. The nuclear DNA content and other morphometric features were evaluated by image analysis (using a modified TICAS system) on 12-microns-thick, Feulgen stained sections. The DNA content of the tumors of both the long-term survivors and the controls varied from the diploid range to highly aneuploid (with a large proportion of the cells having a DNA content above 5N). Overall, the tumors of the controls exhibited a higher ploidy, a greater deviation from the diploid range and a greater variation of nuclear size than did the tumors of the long term survivors. These results suggest that these measurements may be helpful in yielding prognostic information among sets of histologically identical breast tumors of similar pathologic stage. PMID- 2545219 TI - Benzodiazepine receptor ligands. Synthesis and preliminary pharmacological evaluation of 2-aminoalkyl-8-chloro- and 2-aryl-1,2,4-triazolo[3,4-a]phthalazine 3(2H)-ones. AB - Some 2-aminoalkyl-8-chloro- and 2-aryl-1,2,4-triazolo[3,4-a]-phthalazine-3-(2H) ones were synthesized and preliminarily tested in vitro and in vivo as potential benzodiazepine-receptor (BZRs) ligands. 2-Aryl-1,2,4-triazolo[3,4-a]-phthalazine 3(2H)-ones displaced in vitro 3H-diazepam (3H-DZ) from rat brain specific binding sites with Ki (nM) comparable to DZ and chlordiazepoxide used as reference compounds. The specific binding of the triazolones of this study was not enhanced in vitro by 4-aminobutyric acid (GABA) and in vivo they did not show any activity in counteracting the pentylenetetrazole (PTZ) induced convulsions (mice). One of these compounds (IV a) antagonized the effects of DZ in the bicuculline (BIC) induced convulsions test (mice) and the DZ induced muscle relaxant effects in the horizontal wire test. PMID- 2545220 TI - Benzodiazepine receptor ligands. Synthesis and preliminary pharmacological evaluation of some 3,7-disubstituted-4H-[1,2,4]triazino[3,4-a]phthalazines and 3,7-disubstituted-4H-[1,2,4]triazino[3,4-a]phthalazine-4-ones. AB - Some disubstituted triazinophthalazines and traizinophthalazinones were synthesized and tested in vitro for inhibition of the 3H-diazepam (3H-DZ) specific binding to benzodiazepine receptors (BZRs) in membranes from synaptosomes of rat brain and in vivo for their effects on the conditioned behaviour (rat). These compounds showed Ki values (nM) in the in vitro test ranging from 220 to more than 3000 and some of them had approximate ED50S of 30 mg/kg, i.p. (rat) in the conditioned avoidance behaviour test (CR2). PMID- 2545222 TI - Behavioral and physiologic effects of CRH administered to infant primates undergoing maternal separation. AB - Infant rhesus monkeys briefly separated from their mothers emit frequent distress vocalizations and alter their activity levels. These behavioral alterations are accompanied by physiologic changes and activation of the hypothalamic-pituitary adrenal axis. Because corticotropin-releasing hormone (CRH) is a major regulator of the pituitary-adrenal system and has been implicated as a mediator of other aspects of the stress response, we examined the effects of centrally and peripherally administered CRH on the infant monkey's separation response. Intracerebroventricular (ICV) doses less than 10 micrograms did not alter behavior; 10-micrograms doses inhibited behavior without affecting distress vocalizations. The behavioral inhibition did not appear to be due to nonspecific sedation and may be related to increased fearfulness. It was accompanied by increases in adrenocorticotropic hormone (ACTH) and cortisol and small but significant decreases in body temperature and mean arterial pressure. Within an individual animal, no relationship was seen between ICV CRH's effects on physiologic parameters and its effects on behavior. ICV CRH produced significant increases in plasma concentrations of CRH, suggesting the possibility that the effects of CRH were mediated through peripheral mechanisms. However, 10 micrograms of CRH administered intravenously (IV) had no effect on behavior, blood pressure, or body temperature, nor did it increase plasma concentrations of ACTH or cortisol beyond the expected separation-induced elevation. This is interesting because plasma levels of CRH after IV CRH were much greater than after ICV CRH. Our findings suggest that in infant rhesus monkeys undergoing maternal separation, brain CRH systems mediate behavioral inhibition and pituitary-adrenal activation. PMID- 2545221 TI - Benzodiazepine receptor ligands. Synthesis and in vitro binding activity of some 6-substituted-3-aryl-1,2,4-triazolo[3,4-a]phthalazines. AB - Some 3-aryl-6-hydroxymethyl-1,2,4-triazolo[3,4-a]phthalazines (VI a,b) and 3-aryl 6-(2-hydroxyethyl)-1,2,4-triazolo[3,4-a]phthalazines (XVIII) have been synthesized. From (VI b) some 3-aryl-6-(alkylamino)methyl- and 3-aryl-6 (alkoxy)methyl-1,2,4-triazolo[3,4-a]phthalazines have been prepared and evaluated in vitro for their ability to inhibit selective 3H-diazepam (3H-DZ) binding to benzodiazepine-receptors (BZRs) from homogenates of synaptosomes from rat brains. Several compounds have been shown to displace 3H-DZ from BZRs with Ki (nM) values ranging from 2.2 to 88, comparable to those of the reference drugs including Diazepam. PMID- 2545223 TI - Herpes simplex virus type 2 infection of unstimulated human T-lymphocytes. AB - Unstimulated human leukemia T-cell lines (MOLT-4, MT-4) were tested for their susceptibility to herpes simplex virus type 2 (HSV-2) infection. Permissive infection of MT-4 cells was demonstrated by growth curve and infectious center assays. In growth curve experiments new progeny virus replication was detected by 24 hrs and maximum titers of HSV-2 replication were measured by 72 hrs after infection of MT-4 cells, whereas, MOLT-4 cells did not produce detectable infectious HSV-2 in growth curve experiments. It may be that a T-cell subset is involved with infectious HSV-2 production, since 5.7% of MT-4 cells were scored as infectious centers after HSV-2 infection compared to only 0.06% of MOLT-4 cells. Furthermore, HSV-2 infected MT-4 (45% of cells) and MOLT-4 cells (30% of cells) expressed viral induced antigen(s) detected by immunofluorescence assays. These data provide the first evidence of infectious HSV-2 replication in T-cells not prestimulated in vitro with mitogens, pharmacologic agents or growth factors. The establishment of T-cell systems that permit rapid and efficient replication of HSV-2 could greatly facilitate studies on interactions between human herpesviruses and AIDS retroviruses since recent published evidence indicates possible synergistic interactions between these virus groups. PMID- 2545224 TI - Human monoclonal antibodies to influenza virus: IgG subclass and light chain distribution. AB - Three adults and three children were immunized with inactivated or live attenuated influenza vaccines and 98 IgG monoclonal antibodies derived from EBV immortalization of their blood lymphocytes were studied. All antibodies reacted specifically with influenza A H3N2 or H1N1 whole virus and 73 of 74 tested reacted with the purified HA glycoproteins. The majority (76%) of 77 monoclonal antibodies adequately tested by ELISA or solid phase RIA contained lambda light chains. ELISA analysis of the IgG subclass of the six persons' postimmunization anti-influenza serum activity and of monoclonal antibodies derived from them showed a similar predominance of IgG1. PMID- 2545225 TI - Host cell resistance to mouse hepatitis virus type 3 is expressed in vitro in macrophages and lymphocytes. AB - Phenotypic expression of in vivo sensitivity to mouse hepatitis virus type 3 (MHV3) was studied in vitro in macrophages and lymphocytes. MHV3 infections were induced in peritoneal exudate (PE), nonadherent spleen (NAS) and thymus (THY) cells from resistant A/J, susceptible C57BL/6 or semisusceptible (C57BL/6xA/J)F1 mice. Differences in cytopathic effect, cell viability and virus titers were found only at 48 hrs postinfection (p.i.). "Carrier state" infections were performed at 48 hrs p.i. by transfer of supernatants of infected cells to newly collected cells originating from the same strain of mice. A passage-dependent restriction of viral replication was detected in vitro and was expressed in PE, NAS and THY cells as a recessive phenotype. No defective-interfering viral particles were involved in the restriction of viral replication. Results obtained with crossed infections and determination of the number of productively infected cells demonstrated that restriction of viral replication in macrophages and lymphoid cells from resistant A/J mice is controlled by a genetically-determined intrinsic cellular mechanism acting principally on the level of production of infectious viral particles by the infected cell. PMID- 2545226 TI - ELISA for detection of antibodies to a type D retrovirus, SRV-W. AB - An ELISA was developed to detect antibodies to a Type D retrovirus, SRV-W. The interpretation of the ELISA results were based on: 1) Comparisons of known antibody positive (to a closely related type D retrovirus) and negative serum samples, 2) the ability of ELISA reactivity to be absorbed with a Type D virus but not a mock virus preparation, and 3) analysis by a Western Blot assay as an alterative way to identify antibody to the Type D retrovirus. PMID- 2545227 TI - Thyrotrophin-releasing hormone (TRH)-induced growth hormone secretion in fowl: binding of TRH to pituitary membranes. AB - Heat-labile specific binding sites for [3H]3-methylhistidine2-TRH [( 3H]Me-TRH) were demonstrated on chicken adenohypophysial membranes. These binding sites were of both high affinity (dissociation constant, Kd = 15.53 nM) and low capacity (maximum binding capacity, Bmax = 8.73 pmol/g tissue) and of low affinity (Kd = 104.5 nM) and high capacity (Bmax = 32.41 pmol/g). Binding of [3H]Me-TRH to the pituitary membranes was greater at 4 degrees C than at 39 degrees C and occurred with rate constants (k1) of 1.6 x 10(6) and 3.39 x 10(6) M-1 min-1 respectively. Dissociation of [3H]Me-TRH binding at 4 degrees C occurred with a rate constant (k-1) of 0.125 min-1. Binding sites for [3H]Me-TRH were found in the cephalic lobe of the pituitary gland (the location of most lactotrophs and thyrotrophs) and in the caudal lobe (the location of most somatotrophs). The number of binding sites was greater in the caudal lobe than in the cephalic lobe, although the affinity of [3H]Me-TRH binding did not differ. The binding of [3H]Me-TRH to caudal lobe membranes was displaced by Me-TRH, TRH, pGlu-His-Pro-Gly-NH2 and [Glu1]-TRH, with half-maximal effective doses of 33 nM, 70.7 nM, 1.23 microM and 22 microM respectively, but not by [Phe2]-TRH, TRH free acid or His-Pro diketopiperazine. The number of caudal lobe binding sites for [3H]Me-TRH in old birds was less than that in young ones, and the number of binding sites was increased in birds deprived of food for 48 h. TRH-induced GH secretion in birds would thus appear to be mediated by specific receptors on caudal lobe somatotrophs, and these results suggest that physiological changes in GH secretion (during growth and periods of fasting) are causally related to the abundance of TRH-binding sites. PMID- 2545228 TI - Melatonin-binding sites in the rat brain and pituitary mapped by in-vitro autoradiography. AB - Using picomolar concentrations of [125I]iodomelatonin and in-vitro autoradiography, specific melatonin-binding sites have been mapped in the rat brain and pituitary. Using this same technique, high-affinity melatonin receptors had previously been identified in the suprachiasmatic nucleus (SCN) and median eminence regions of the rat hypothalamus. The presence of melatonin binding in the SCN has been confirmed, but the second area of binding has been identified as the pars tuberalis of the pituitary, and a completely novel area of binding is also reported in the area postrema. The existence of lower affinity melatonin receptors in the rat brain was also investigated using in-vitro autoradiography and higher concentrations of [125I]iodomelatonin. No further sites of specific binding were, however, disclosed. PMID- 2545229 TI - The Hodgkin-associated Ki-1 antigen exists in an intracellular and a membrane bound form. AB - The Hodgkin-associated Ki-1 antigen occurs in two different molecular forms. The 120-kDa membrane-associated form is a phosphorylated glycoprotein, which is derived from a non-phosphorylated intracellular 84-kDa apoprotein that is co translationally N-glycosylated with a carbohydrate portion of 6 kDa. The other form of the Ki-1 antigen is a non-glycosylated phosphoprotein of 57 kDa which only occurs intracellularly. Both forms of the antigen are phosphorylated at serine residues. Enzymatic cleavage with sialidase reduced the 120-kDa membrane antigen by about 15 kDa, while its 90-kDa precursor and the 57-kDa intracellular form of the Ki-1 antigen remained unaltered. Pulse-chase experiments revealed that the 57-kDa and 90/120-kDa molecules are synthesized independently of each other. Four to eight hours after synthesis, the degradation of the 120-kDa molecule to a 105-kDa membrane-associated intermediate begins. This is further processed and appears in the cell supernate as a 90-kDa molecule. Hodgkin's disease-derived, Epstein-Barr virus-transformed cell lines and the acute T cell leukemia line MOLT-4 contain both forms of the Ki-1 antigen, whereas only the 57 kDa intracellular antigen is expressed in U266/B1 myeloma cells, in the Burkitt lymphoma cell lines Raji and Daudi and in acute promyelocytic HL-60 leukemia cells. PMID- 2545230 TI - Suggested links between different types of dementias: Creutzfeldt-Jakob disease, Alzheimer disease, and retroviral CNS infections. AB - Several lines of investigation identify common features of Creutzfeldt-Jakob disease, Alzheimer disease, and retroviral CNS infections. We discuss salient neuropathological, genetic, transmission, and transformation properties that suggest there may be common pathways in the pathogenesis of dementias. We also briefly present potential retroviral mechanisms that may be implicated in all of these dementias. PMID- 2545231 TI - Chromogranins--new sensitive markers for neuroendocrine tumors. AB - Chromogranins A, B and C, proteins that are costored and coreleased with peptides and amines, have been identified in a variety of endocrine and nervous tissues, both normal and neoplastic. We examined the secretion of chromogranin A and chromogranin A + B by hormone-producing tumors in patients with endocrine pancreatic tumors (EPT), carcinoid tumors, pheochromocytomas and small cell lung cancer (SCLC). Radioimmunoassay (RIA) of the plasma/serum concentrations of chromogranin A + B showed a greater sensitivity than RIA of chromogranin A alone. All patients with EPT, carcinoids and pheochromocytomas had increased levels of chromogranin A + B, whereas a small number of the patients (5/18 with EPT and 1/3 with pheochromocytomas) had normal levels of chromogranin A. Also in immunocytochemical stainings, our polyclonal antiserum detecting both chromogranin A and B showed a greater sensitivity than other available antisera against chromogranin A, B and C. PMID- 2545232 TI - Platelet-derived growth factor. Three isoforms that bind to two distinct cell surface receptors. AB - Several isoforms of the platelet-derived growth factor (PDGF) have been identified and purified to date. The functional effects of the PDGF isoforms differ, due to their different interactions with at least two distinct cell surface receptors. Thus, PDGF constitutes an example of an expanding growth factor family, characterized by possibilities for fine-tuned regulation of action. In this manuscript, we describe the structural features of the PDGF isoforms as well as their receptors. PMID- 2545233 TI - Surgical treatment of carcinoids and endocrine pancreatic tumours. AB - The common association of neuroendocrine abdominal neoplasms, carcinoids and endocrine pancreatic tumours, with often severe endocrine symptoms has justified considerable efforts to improve treatment in these conditions. Surgery still constitutes the principle therapy for the majority of these tumours. However, the introduction of new means of medical treatment with cytostatic agents, or more recently interferon and a somatostatin analogue, seem to have impact on indications and the extent of surgery in malignant forms of these tumours. It is thus probable that a surgical tumour reduction will increase the possibilities to achieve positive effects of the medical treatment even in advanced malignancies. However, the existence of an alternative medical therapy has increased obligations that surgery should be performed without morbidity or mortality. PMID- 2545234 TI - The viral/rickettsial diseases laboratory. PMID- 2545235 TI - Observations on the effects of two ceramics on the strength of bone subjected to cryosurgery. AB - Implants of dense hydroxyapatite and beta tricalcium phosphate were implanted over rat mandibular bone, or rat mandibular bone previously subjected to cryosurgery, which is known to produce a reduction in strength. After selected intervals, up to 6 months, the bone was stress-tested to assess fracture strength. Both materials demonstrated an effect in off-setting strength loss after cryosurgery and these findings are discussed. PMID- 2545236 TI - Site-directed mutagenesis of lysine 58 in a putative ATP-binding domain of the calmodulin-sensitive adenylate cyclase from Bordetella pertussis abolishes catalytic activity. AB - A 2.7-kb cya A gene fragment encoding the amino-terminal end of the calmodulin sensitive adenylate cyclase from Bordetella pertussis has been placed under the control of the lac promoter for expression in Escherichia coli. Following induction with isopropyl beta-D-thiogalactoside, calmodulin-sensitive adenylate cyclase activity was detected in a cell extract from E. coli. The expression vector directed the synthesis of a 90-kDa polypeptide that was recognized by rabbit polyclonal antibodies raised against the catalytic subunit of B. pertussis adenylate cyclase. Inspection of the deduced amino acid sequence of the cya A gene product revealed a sequence with homology to consensus sequences for an ATP binding domain found in many ATP-binding proteins. On the basis of the analysis of nucleotide binding proteins, a conserved lysine residue has been implicated in the binding of ATP. A putative ATP-binding domain in the B. pertussis adenylate cyclase possesses an analogous lysine residue at position 58. To test whether lysine 58 of the B. pertussis adenylate cyclase is a crucial residue for enzyme activity, it was replaced with methionine by oligonucleotide-directed mutagenesis. E. coli cells were transformed with the mutant cya A gene, and the expressed gene product was characterized. The mutant protein exhibited neither basal nor calmodulin-stimulated enzyme activity, indicating that lysine 58 plays a critical role in enzyme catalysis. PMID- 2545237 TI - Photoreceptor channel activation by nucleotide derivatives. AB - Cyclic nucleotide activated sodium currents were recorded from photoreceptor outer segment membrane patches. The concentration of cGMP and structurally similar nucleotide derivatives was varied at the cytoplasmic membrane face; currents were generated at each concentration by the application of a voltage ramp. Nucleotide-activated currents were analyzed as a function of both concentration and membrane potential. For cGMP, the average K0.5 at 0 mV was 24 microM, and the activation was cooperative with an average Hill coefficient of 2.3. Of the nucleotide derivatives examined, only 8-[[(fluorescein-5-yl carbamoyl)methyl]thio]-cGMP (8-Fl-cGMP) activated the channel at lower concentrations than cGMP with a K0.5 of 0.85 microM. The next most active derivative was 2-amino-6-mercaptopurine riboside 3',5'-monophosphate (6-SH-cGMP) which had a K0.5 of 81 microM. cIMP and cAMP had very high K0.5 values of approximately 1.2 mM and greater than 1.5 mM, respectively. All nucleotides displayed cooperativity in their response and were rapidly reversible. Maximal current for each derivative was compared to the current produced at 200 microM cGMP; only 8-Fl-cGMP produced an identical current. The partial agonists 6-SH cGMP, cIMP, and cAMP activated currents which were approximately 90%, 80%, and 25% of the cGMP response, respectively. 5'-GMP, 2-aminopurine riboside 3',5' monophosphate, and 2'-deoxy-cGMP produced no detectable current. The K0.5 values for cGMP activation, examined from -90 to +90 mV, displayed a weak voltage dependence of approximately 400 mV/e-fold; the index of cooperativity was independent of the applied field.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545238 TI - Characterization of the helicase activity of the Escherichia coli RecBCD enzyme using a novel helicase assay. AB - We describe an assay to measure the extent of enzymatic unwinding of DNA by a DNA helicase. This assay takes advantage of the quenching of the intrinsic protein fluorescence of Escherichia coli SSB protein upon binding to ssDNA and is used to characterize the DNA unwinding activity of recBCD enzyme. Unwinding in this assay is dependent on the presence of recBCD enzyme and linear dsDNA, is consistent with the known properties of recBCD enzyme, and closely parallels other methods for measuring recBCD enzyme helicase activity. The effects of varying temperature, substrate concentrations, enzyme concentration, and mono- and divalent salt concentrations on the helicase activity of recBCD enzyme were characterized. The apparent Km values for recBCD enzyme helicase activity on linear M13 dsDNA molecules at 25 degrees C are 0.6 nM dsDNA molecules and 130 microM ATP, respectively. The apparent turnover number for unwinding is approximately 15 microM base pairs s-1 (microM recBCD enzyme)-1. When this rate is corrected for the observed stoichiometry of recBCD enzyme binding to dsDNA, kcat for helicase activity corresponds to an unwinding rate of approximately 250 base pairs of DNA s-1 (functional recBCD complex)-1 at 25 degrees C. At 37 degrees C, the apparent Km value for dsDNA molecules was the same as that at 25 degrees C, but the apparent turnover number became 56 microM base pairs s-1 (microM recBCD enzyme)-1 [or 930 base pairs s-1 (functional recBCD complex)-1 when corrected for observed stoichiometry]. With increasing NaCl concentration, kcat peaks at 100 mM, and the apparent Km value for dsDNA increases by 3-fold at 200 mM NaCl. In the presence of 5 mM calcium acetate, the apparent Km value is increased by 3-fold, and kcat decreased by 20-30%. We have also shown that recBCD enzyme molecules are able to catalytically unwind additional dsDNA substrates subsequent to initiation, unwinding, and dissociation from a previous dsDNA molecule. PMID- 2545240 TI - Dynamics of proton diffusion within the hydration layer of phospholipid membrane. AB - The diffusion of protons at the immediate vicinity of (less than 10 A from) a phospholipid membrane is studied by the application of the laser-induced proton pulse. A light-sensitive proton emitter (8-hydroxypyrene-1,3,6-trisulfonate) was trapped exclusively in the hydration layers of multilamellar vesicles made of egg phosphatidylcholine, and the protons were dissociated by a synchronizing laser pulse. The recombination of the proton with pyranin anion was monitored by time resolved spectroscopy and analyzed by a diffusion-controlled formalism. The measured diffusion coefficient is only slightly smaller than the diffusion coefficient of proton in bulk water. Modulating the width of the hydration layer by external pressure had a direct effect on the diffusibility of the proton: the narrower the hydration layer, the slower is the diffusion of protons. PMID- 2545239 TI - Characterization of the adenosinetriphosphatase activity of the Escherichia coli RecBCD enzyme: relationship of ATP hydrolysis to the unwinding of duplex DNA. AB - We find that the rate of dsDNA-dependent ATPase activity is biphasic, with a fast component which represents the unwinding of the dsDNA and a slow component which results from the ssDNA-dependent ATPase activity of recBCD enzyme. Comparison of the ATPase and helicase activities permits evaluation of the efficiency of ATP hydrolysis during unwinding. This efficiency can be calculated from the maximum rates of ATPase and helicase activities and is found to range between 2.0 and 3.0 ATP molecules hydrolyzed per base pair of DNA unwound. The number of ATP molecules hydrolyzed per base pair unwound is not altered by temperature but does increase at low concentrations of DNA and high concentrations of sodium chloride and magnesium acetate. The apparent Km values for the DNA and ATP substrates of recBCD enzyme dsDNA-dependent ATPase activity at 25 degrees C were determined to be 0.13 nM DNA molecules and 85 microM ATP, respectively. The observed kcat value is approximately 45 microM ATP s-1 (microM recBCD enzyme)-1. If this rate is corrected for the measured stoichiometry of recBCD enzyme binding to dsDNA, the kcat for ATPase activity corresponds to an ATP hydrolysis rate of approximately 740 ATP molecules s-1 (functional recBCD complex)-1 at 25 degrees C. PMID- 2545241 TI - Photoaffinity labeling by [3H]-N5-methyl-N5-isobutylamiloride of proteins which cofractionate with Na+/H+ antiport activity. AB - N5-Methyl-N5-isobutylamiloride (MIA) is one of a series of 5-N-substituted amiloride analogues which exhibit high affinity and specificity for inhibition of Na+/H+ antiport. Amiloride-sensitive [3H]MIA binding to renal brush border membranes exhibited a Kd of 250 nM and a Bmax of 8.6 pmol/mg of protein. Specific binding was optimal at pH 7.5 and inhibited in the presence of Na+ and Li+. Inhibition by amiloride exhibited biphasic kinetics. After resolution of solubilized membranes by high-pressure liquid chromatography, MIA binding activity cofractionated together with Na+/H+ antiport activity, measured after reconstitution in asolectin vesicles, into a major and a minor peak. When fractions containing the major peak of Na+/H+ antiport activity were incubated with [3H]MIA and then photolyzed with a mercury arc lamp, covalent incorporation of label into polypeptides of apparent molecular mass 81 and 107 kDa was observed. These photolabeled bands were also observed in intact brush border membranes in addition to labeled polypeptides of apparent molecular mass 60 and 46 kDa, respectively. Labeling was inhibited by amiloride, reduced in the presence of Na+, and not observed in the absence of photolysis. These data point to the 81- and 107-kDa polypeptides as candidates for identification as components of a Na+/H+ antiport system in renal brush border membranes. PMID- 2545243 TI - Myeloperoxidase-mediated inhibition of microbial respiration: damage to Escherichia coli ubiquinol oxidase. AB - A microbicidal system, mediated by neutrophil myeloperoxidase, inhibits succinate dependent respiration in Escherichia coli at rates that correlate with loss of microbial viability. Succinate dehydrogenase, the initial enzyme of the succinate oxidase respiratory pathway, catalyzes the reduction of ubiquinone to ubiquinol, which is reoxidized by terminal oxidase complexes. The steady-state ratio of ubiquinol to total quinone (ubiquinol + ubiquinone) reflects the balance between dehydrogenase-dependent ubiquinone reduction and terminal oxidase-dependent ubiquinol oxidation. Myeloperoxidase had no effect on total quinone content of E. coli but altered the steady-state ratio of ubiquinol to total quinone. The ratio doubled for organisms incubated with the myeloperoxidase system for 10 min, suggesting decreased ubiquinol oxidase activity, which was confirmed by observation of a 50% decrease in oxidation of the ubiquinol analogue 2,3 dimethoxy-5-methyl-6-decyl-1,4-benzoquinol. Despite inhibition of ubiquinol oxidase, overall succinate oxidase activity remained unchanged, suggesting that succinate dehydrogenase activity was preserved and that the dehydrogenase was rate limiting. Microbial viability was unaffected by early changes in ubiquinol oxidase activity. Longer (60 min) exposure of E. coli to the myeloperoxidase system resulted in only modest further inhibition of the ubiquinol oxidase, but the ubiquinol to total quinone ratio fell to 0%, reflecting complete loss of succinate dehydrogenase activity. Succinate oxidase activity was abolished, and there was extensive loss of microbial viability. Early myeloperoxidase-mediated injury to ubiquinol oxidase appeared to be compensated for by higher steady-state levels of ubiquinol which sustained electron turnover by mass effect. Later myeloperoxidase-mediated injuries eliminated succinate-dependent ubiquinone reduction, through inhibition of succinate dehydrogenase, with loss of succinate oxidase activity, effects which were associated with, although not clearly causal for, microbicidal activity. PMID- 2545242 TI - Nucleophilic behavior of lysine-501 of the alpha-polypeptide of sodium and potassium ion activated adenosinetriphosphatase consistent with a role in binding adenosine triphosphate. AB - An immunoadsorbent specific for the carboxy-terminal sequence -GAPER, which comprises residues 502-506 of the alpha-polypeptide of ovine sodium and potassium ion activated adenosinetriphosphatase [(Na+ + K+)-ATPase], was used to isolate the products of the reaction between the lysine immediately preceding this sequence in the intact protein and either [3H]acetic anhydride or fluorescein 5' isothiocyanate. Changes in the apparent nucleophilicity of this lysine, Lys501, were observed with both reagents when ATP was bound by the intact, native enzyme poised in the E1 conformation or when the structure of the enzyme was changed from the E1 conformation into the E2-P conformation. With both reagents, a decrease of more than 4-fold in the yield of incorporation occurred during the former change, but a decrease of only 2-fold occurred during the latter. Because a much larger decrease occurred when ATP was bound in the absence of a conformational change than occurred when a major conformational change took place in the absence of the occupation of the active site, these changes in the incorporation of [3H]acetyl suggest that Lys501 from the alpha polypeptide is directly involved in binding ATP within the active site of (Na+ + K+)-ATPase. The immunochemical reactions between the specific polyclonal antibodies raised against the sequence-GAPER and denatured or enzymically active (Na+ + K+)-ATPase were also investigated. Western blots and the inhibition of enzymic activity caused by the antibody have shown that it can bind to both the denatured and the native form of the alpha-polypeptide, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545244 TI - On the inhibition of [Na+,K+]-ATPases by the components of Naja mossambica mossambica venom: evidence for two distinct rat brain [Na+,K+]-ATPase activities. AB - We have compared the effects of highly purified preparations of cardiotoxins and phospholipases A2 from Naja mossambica mossambica venom on rat brain [Na+,K+] ATPase activity. The results were the following: (i) micromolar concentrations of cardiotoxin preparations were required to inhibit [Na+,K+]-ATPase activity to the extent achieved by picomolar concentrations of phospholipases A2; i.e., the inhibitory effect of cardiotoxins appeared to be related to the contamination of the preparations by trace amounts of phospholipase A2; (ii) comparing phospholipases A2 from varied origins, a correlation was observed between [Na+,K+]-ATPase inhibition, isoelectric point, and toxicity for mice; (iii) when rat brain membranes were used, incubation for extended times with the most basic N. mossambica mossambica phospholipase A2 resulted in a biphasic [Na+,K+]-ATPase inhibition, suggesting that two distinct [Na+,K+]-ATPases were affected differentially. In contrast, incubation of rat brain membranes with either porcine pancreatic phospholipase A2, notexin, or beta-bungarotoxin and also incubation of erythrocyte membranes with the most basic N. mossambica mossambica phospholipase A2 produced monophasic [Na+,K+]-ATPase inhibitions. We discuss a possible specific action of toxic, basic phospholipase A2 on one of the [Na+,K+] ATPase isoforms of excitable membranes. PMID- 2545245 TI - Characterization of the cytochrome c oxidase in isolated and purified plasma membranes from the cyanobacterium Anacystis nidulans. AB - Functionally intact plasma membranes were isolated from the cyanobacterium (blue green alga) Anacystis nidulans through French pressure cell extrusion of lysozyme/EDTA-treated cells, separated from thylakoid membranes by discontinuous sucrose density gradient centrifugation, and purified by repeated recentrifugation. Origin and identity of the chlorophyll-free plasma membrane fraction were confirmed by labeling of intact cells with impermeant protein markers, [35S]diazobenzenesulfonate and fluorescamine, prior to membrane isolation. Rates of oxidation of reduced horse heart cytochrome c by purified plasma and thylakoid membranes were 90 and 2 nmol min-1 (mg of protein)-1, respectively. The cytochrome oxidase in isolated plasma membranes was identified as a copper-containing aa3-type enzyme from the properties of its redox-active and EDTA-resistant Cu2+ ESR signal, the characteristic inhibition profile, reduced minus oxidized difference spectra, carbon monoxide difference spectra, photoaction and photodissociation spectra of the CO-inhibited enzyme, and immunological cross-reaction of two subunits of the enzyme with antibodies against subunits I and II, and the holoenzyme, of Paracoccus denitrificans aa3 type cytochrome oxidase. The data presented are the first comprehensive evidence for the occurrence of aa3-type cytochrome oxidase in the plasma membrane of a cyanobacterium similar to the corresponding mitochondrial enzyme (EC 1.9.3.1). PMID- 2545246 TI - Resonance Raman evidence that distal histidine protonation removes the steric hindrance to upright binding of carbon monoxide by myoglobin. AB - The resonance Raman band assigned to Fe--CO stretching in the sperm whale myoglobin CO adduct shifts from 507 cm-1 at neutral pH to 488 cm-1 at low pH, in concert with a shift of the C-O stretching infrared band from 1947 to 1967 cm-1 (Fuchsman & Appleby, 1979), while the 575-cm-1 Fe-C-O bending RR band loses intensity. The pKa that characterizes these changes is approximately 4.4. The vibrational frequencies at low pH are well modeled by the protein-free CO, imidazole adduct of protoheme in a nonpolar solvent while those at high pH are modeled by the adduct of a heme with a covalent strap (Yu et al., 1983) which inhibits upright CO binding. It is inferred that the Fe-C-O unit changes from a tilted to an upright geometry when the distal histidine is protonated, because its side chain swings out of the heme pocket due to electrostatic repulsion with a nearby arginine residue. A different protonation step (pKa = 5.7), which has been shown to modulate the CO rebinding kinetics (Doster et al., 1982) as well as the optical spectrum (Fuchsman & Appleby, 1979), is suggested to involve a global structure change associated with protonation of histidine residues distant from the heme. PMID- 2545247 TI - Cytochrome c oxidase: evidence for interaction of water molecules with cytochrome a. AB - The resonance Raman spectra of cytochrome c oxidase in protonated buffer compared to that in deuterated buffer indicate that water molecules are near the heme of cytochrome a. Differences in widths of the heme line at 1610 cm-1, after short exposure to D2O, and, additionally, of the heme line at 1625 cm-1, after long exposure, can be accounted for by changes in resonance vibrational energy transfer between modes of cytochrome a2+ and the bending mode of water molecules in the heme pocket. On the basis of the assignment of these modes, we place one water molecule near the vinyl group and one water molecule near the formyl group of the cytochrome a heme. These water molecules may play several possible functional roles. PMID- 2545248 TI - G protein-effector coupling: binding of rod phosphodiesterase inhibitory subunit to transducin. AB - The cyclic GMP phosphodiesterase of retinal rods is composed of three distinct polypeptides: alpha (90 kDa), beta (86 kDa), and gamma (10 kDa). In this multimeric form, the enzyme is inhibited. Its activity is stimulated by the interaction with the GTP-bound form of the T alpha subunit of transducin and reversed upon the recombination of the inhibitory gamma subunit with the catalytic alpha beta subunit. We show here by a novel coimmunoprecipitation technique that the gamma subunit, but not the alpha beta subunit, forms a 1:1 complex with T alpha. The binding of gamma to T alpha is nucleotide-dependent and is facilitated by GTP gamma S or Gpp(NH)p. This study provides convincing evidence that the T alpha-GTP subunit of transducin stimulates phosphodiesterase activity by binding to gamma and physically carrying it away from alpha beta. PMID- 2545249 TI - Mutation-induced perturbation of the cytochrome c alkaline transition. AB - The possible influence of residue Phe-82 in the cytochrome c alkaline isomerization has been evaluated by spectrophotometric pH titrations of a family of mutant yeast iso-1-cytochromes c in which the identity of the residue at this position has been varied. The pKa for the exchange of the Met-80 heme iron ligand was determined from pH titrations in which the S----Fe charge-transfer band (695 nm) was monitored and was found to be 8.5 for the wild type, 7.7 for Ser-82, 7.7 for Gly-82, 7.2 for Leu-82, and 7.2 for Ile-82. pH-jump experiments [Davis et al. (1974) J. Biol. Chem. 249, 2624] established that substitutions at position 82 affect the alkaline isomerization by lowering the pKa of the titrating group by as much as 1.4 pK units; for the Ser-82 and Gly-82 variants, there is also a small effect on the Keq for the ligand exchange equilibrium. On the basis of these findings, we conclude that one critical role for Phe-82 in the wild-type protein is stabilization of the native heme binding environment. PMID- 2545250 TI - Intrinsic uncoupling in proton-pumping cytochrome c oxidase: pH dependence of cytochrome c oxidation in coupled and uncoupled phospholipid vesicles. AB - The pH dependence of the transient aerobic kinetics of cytochromes c and a has been investigated with cytochrome oxidase reconstituted in phospholipid vesicles in the absence and presence of an uncoupler and an ionophore. The cytochrome a reduction level immediately after the burst phase was 60-80% and was not significantly changed by the addition of uncoupler and/or ionophore. The coupled rate of ferro-cytochrome c oxidation increases linearly with decreasing pH in the range 8.4-5.4. The increase in rate on uncoupling becomes less with decreasing pH and low cytochrome c concentration, being almost zero at pH 5.4. The coupled rate is increased by a lowering of the outside pH when the inside pH is constant. Varying the inside pH with a constant outside pH of 7.4 has little effect on the rate. It is suggested that the electrochemical potential has two separate effects on the coupled rate: the pH gradient mainly slows down the intramolecular electron transfer, but the membrane potential also lowers the second-order rate constant for the reaction with cytochrome c. The results are interpreted in terms of a model in which protonation of an acid-base group with a pKa of 6.4 from the inside increases the catalytic constant. Protonation from the outside, on the other hand, leads to an intrinsic uncoupling, because the protonated enzyme in the output state can return to the input state. This has no adverse physiological effect, since it becomes significant only at pH values well below 7. PMID- 2545251 TI - Evidence for the presence of a carbohydrate moiety in fluorescein isothiocyanate labeled fragments of rat gastric (H+-K+)-ATPase. AB - Limited tryptic digestion of fluorescein isothiocyanate (FITC)-labeled (H+-K+) ATPase from rat resting light gastric membranes produced a soluble 27-kDa polypeptide which retained the fluorescence of the parent enzyme. Its production was markedly enhanced in the presence of an amphiphilic detergent, Zwittergent 3 14, which potently inhibits the ATPase activity. This increase is probably due to protection of certain tryptic cleavage sites through conformational changes of the membrane enzyme by the detergent. The NH2-terminal sequence of the 27-kDa polypeptide corresponded exactly to that beginning at Asn-369 of the cDNA-deduced primary structure of the rat ATPase. The presence of the phosphorylation site, Asp-385, and FITC-labeled Lys-517, which is known to be a part of the ATP-binding site, indicates that the 27-kDa polypeptide contains a major cytoplasmic portion of (H+-K+)-ATPase. Interestingly, the polypeptide was stained with periodate Schiff's base, indicating its glycoprotein nature. The carbohydrate group attached to the polypeptide seems to include at least an N-linked high-mannose moiety, since the polypeptide showed Con A binding activity as detected with a Con A-biotin/avidin-peroxidase assay on nitrocellulose transblots. Also, its Con A binding activity was inhibited by excess methyl alpha-D-mannopyranoside and disappeared upon treatment of the polypeptide with endoglycosidase H and N glycanase. Further tryptic action converted the 27-kDa polypeptide to 2 smaller FITC-labeled polypeptides of 25 and 15 kDa, which lost 18 and 96 amino acid residues, respectively, from the NH2 terminus of the parent polypeptide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545253 TI - Sequence specificity of DNA cleavage by bis(1,10-phenanthroline)copper(I): effects of single base pair transitions on the cleavage of preferred pyrimidine purine-pyrimidine triplets. AB - The cleavage of DNA restriction fragments by bis(1,10-phenanthroline)copper(I) [[(OP)2CuI]+] is sequence dependent: the trimer TAT is most strongly preferred, while the trimer TGT and tetramers TAAT, TAGT, and CAGT are strongly to moderately preferred [Veal, J. M., & R. L. (1988) Biochemistry 27, 1822-1827]. [(OP)2CuI]+ cleavage of a series of oligonucleotide duplexes of the type 5' CCCTPyPuPyCCCC-3'/3'-GGGAPuPyPuGGGG-5' (Py = pyrimidine; Pu = purine) was examined to determine the effects of purine substituents in the central triplet on specificity. The relative cleavage rates of different PyPuPy triplets in oligomers were similar to those observed for restriction fragments. The undecamer duplex containing the trimer TAT (TTATC) was most preferentially cleaved, predominantly at the central adenosine and the adjacent 3'-thymidine. Duplexes differing from TTATC by a single A.T----G.C transition in the central triplet were cleaved at significantly reduced rates relative to TTATC, the order of preference being TAT greater than TGT greater than TAC greater than CAT. By contrast, duplexes differing from TTATC by a single A.T----I.C transition were cleaved at rates similar to those for TTATC when the transition occurred at the 5'-pyrimidine or central purine [i.e., C(.I)AT and TIT]. A duplex containing the trimer TAC(.I) was cleaved at a reduced rate similar to the duplex containing TAC(.G). The guanine 2-amino group at positions 1 and 2, but not position 3, of a 5'-PyPuPy-3' trimer is therefore implicated as a strong inhibitor of DNA binding by the copper-phenanthroline complex.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545252 TI - Role of arginine-38 in regulation of the cytochrome c oxidation-reduction equilibrium. AB - Arg-38 is an internal residue of mitochondrial cytochrome c that is close to heme propionate-7. Previous work comparing the behavior of cytochromes c from several species [Moore, G. R., Harris, D. E., Leitch, F. A., & Pettigrew, G. W. (1984) Biochim. Biophys. Acta 764, 331-342] has suggested that Arg-38 lowers the pKa of this propionate group and thereby accounts for the relative pH independence of the cytochrome c reduction potential from pH 5 to pH 8. The influence of Arg-38 on the oxidation-reduction equilibrium of yeast iso-1-cytochrome c has now been investigated by electrochemical, NMR, and theoretical analysis of six specifically mutated forms of this protein in which Arg has been replaced by Lys, His, Gln, Asn, Leu, or Ala. As the electron-withdrawing character of the residue at position 38 decreases, the reduction potential of the protein also decreases, with the largest decrease (ca. 50 mV) observed for the Ala variant. However, the variation in the reduction potentials of the mutants as a function of pH was similar to that observed for the wild-type protein. The effects of some of these mutations on the pKa values of His-33 and His-39 have been determined by NMR spectroscopy and found to be minimal. Calculations of the electrostatic free energy for the Leu-38 variant predict a decrease in the reduction potential of this mutant that is remarkably close to that observed experimentally. This work establishes that while Arg-38 contributes to the relatively high reduction potential of cytochrome c, this residue does not appear to be the sole functionality responsible for lowering the heme propionate-7 pKa. PMID- 2545254 TI - T7 RNA polymerase interacts with its promoter from one side of the DNA helix. AB - The interactions of T7 RNA polymerase with its promoter DNA have been previously probed in footprinting experiments with either DNase I or (methidiumpropyl-EDTA) Fe(II) to cleave unprotected DNA [Basu, S., & Maitra, U. (1986) J. Mol. Biol. 190, 425-437. Ikeda, R. A., & Richardson, C. C. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 3614-3618]. Both of these reagents have drawbacks; DNase I is a bulky reagent and so provides low resolution, and (methidiumpropyl-EDTA)-Fe(II) intercalates into DNA and is therefore biased toward cleavage of double-stranded DNA. In this study, the interaction between the polymerase and the promoter has been probed with Fe(II)-EDTA. This reagent generates reactive hydroxyl radicals free in solution, which produces a more detailed picture of the polymerase promoter complex. Two protected regions are observed on each of the two promoter DNA strands: from position -17 to position -13 and from position -7 to position 1 on the coding strand and from position -14 to position -9 and from position -3 to position +2 on the noncoding strand. From this pattern it is clear that if recognition occurs via double-stranded B-form DNA, then the protected regions lie on one face of the DNA helix, and therefore the enzyme must interact predominantly from one side of the DNA helix. Digestion of the DNA in a polymerase-promoter complex with a single-strand-specific endonuclease shows that a small region of the noncoding strand near position -5 is susceptible to cleavage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545255 TI - Quantitative footprinting analysis using a DNA-cleaving metalloporphyrin complex. AB - The results of quantitative footprinting studies involving the antiviral agent netropsin and a DNA-cleaving cationic metalloporphyrin complex are presented. An analysis of the footprinting autoradiographic spot intensities using a model previously applied to footprinting studies involving the enzyme DNase I [Ward, B., Rehfuss, R., Goodisman, J., & Dabrowiak, J. C. (1988) Biochemistry 27, 1198 1205] led to very low values for netropsin binding constants on a restriction fragment from pBR-322 DNA. In this work, we show that, because the porphyrin binds with high specificity to DNA, it does not report site loading information in the same manner as does DNase I. We elucidate a model involving binding equilibria for individual sites and include competitive binding of drug and porphyrin for the same site. The free porphyrin and free drug concentrations are determined by binding equilibria with the carrier (calf thymus DNA) which is present in excess and acts as a buffer for both. Given free porphyrin and free netropsin concentrations for each total drug concentration in a series of footprinting experiments, one can calculate autoradiographic spot intensities in terms of the binding constants of netropsin to the various sites on the 139 base pair restriction fragment. The best values of these binding constants are determined by minimizing the sum of the squared differences between calculated and experimental footprinting autoradiographic spot intensities. Although the determined netropsin binding constants are insensitive to the value assumed for the porphyrin binding constant toward its highest affinity sites, the best mean square deviation between observed and calculated values, D, depends on the choice of (average) drug binding constant to carrier DNA, Kd.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545256 TI - Conformation and dynamics of a left-handed Z-DNA hairpin: studies of d(CGCGCGTTTTCGCGCG) in solution. AB - The physical properties of the DNA oligomer d(CGCGCGTTTTCGCGCG) in solvents containing 4 M NaClO4 and 0.1 M NaCl were investigated by proton NMR, optical melting, and circular dichroism spectroscopy. Results of these investigations are as follows: (i) The DNA hexadecamer exists as a unimolecular hairpin in either high or low salt. (ii) In high salt the stem region of the hairpin is in the left handed Z conformation. (iii) In either high or low salt, the duplex stem of the hairpin is stabilized against melting by approximately 40 degrees C compared to the linear core duplex. The added stability of the hairpin is entropic in origin. (iv) In high salt, as the temperature is elevated, the equilibrium structure of the duplex stem of the hairpin shifts from the Z to the B conformation before melting. (v) In low salt, when the DNA duplex exists in the B conformation, attachment of a T4 single-strand loop to one end only slightly decreases (by 14%) the correlation time of the CH5-CH6 interproton vector. In high salt, when the DNA duplex exists in the Z conformation, the correlation time of the CH5-CH6 interproton vector decreases by 51%. Since these viscosity-corrected correlation times are taken to be indicators of duplex motions on the nanosecond time scale, this result directly suggests a larger amplitude of these motions is present in the duplex stem of the hairpin when it exists in the Z conformation. PMID- 2545257 TI - Magnetic resonance spectral characterization of the heme active site of Coprinus cinereus peroxidase. AB - Examination of the peroxidase isolated from the inkcap Basidiomycete Coprinus cinereus shows that the 42,000-dalton enzyme contains a protoheme IX prosthetic group. Reactivity assays and the electronic absorption spectra of native Coprinus peroxidase and several of its ligand complexes indicate that this enzyme has characteristics similar to those reported for horseradish peroxidase. In this paper, we characterize the H2O2-oxidized forms of Coprinus peroxidase compounds I, II, and III by electronic absorption and magnetic resonance spectroscopies. Electron paramagnetic resonance (EPR) and nuclear magnetic resonance (NMR) studies of this Coprinus peroxidase indicate the presence of high-spin Fe(III) in the native protein and a number of differences between the heme site of Coprinus peroxidase and horseradish peroxidase. Carbon-13 (of the ferrous CO adduct) and nitrogen-15 (of the cyanide complex) NMR studies together with proton NMR studies of the native and cyanide-complexed Coprinus peroxidase are consistent with coordination of a proximal histidine ligand. The EPR spectrum of the ferrous NO complex is also reported. Protein reconstitution with deuterated hemin has facilitated the assignment of the heme methyl resonances in the proton NMR spectrum. PMID- 2545259 TI - mu-conotoxin GIIIA, a peptide ligand for muscle sodium channels: chemical synthesis, radiolabeling, and receptor characterization. AB - The peptide conotoxin GIIIA from Conus geographus L. venom, which specifically blocks sodium channels in muscle, has been synthesized by a solid-phase method. The three disulfide bridges were formed by air oxidation. After HPLC purification, the synthetic product was shown to be identical with the native conotoxin GIIIA from Conus geographus. A high specific activity, 125I derivative of mu-conotoxin was prepared and used for binding assays to the Na channel from Electrophorus electric organ. Specific binding could be abolished by competition with tetrodotoxin. The radiolabeled toxin was specifically cross-linked to the Na channel. These studies demonstrate that mu-conotoxin GIIIA can be used to define the guanidinium toxin binding site and will be a useful ligand for understanding functionally important differences between Na channel subtypes. PMID- 2545260 TI - Structure of a hydroxyl radical induced DNA-protein cross-link involving thymine and tyrosine in nucleohistone. AB - Hydroxyl radical induced formation of a DNA-protein cross-link involving thymine and tyrosine in nucleohistone is described. Hydroxyl radicals were generated in N2O-saturated aqueous solution by ionizing radiation. Samples of nucleohistone were hydrolyzed with HCl and trimethylsilylated. Analysis of irradiated samples by gas chromatography-mass spectrometry with selected-ion monitoring showed the presence of a thymine-tyrosine cross-link on the basis of typical fragment ions from the previously known mass spectrum of its trimethylsilyl derivative. The yield of this DNA-protein cross-link in nucleohistone was measured at incrementing doses of radiation and found to be a linear function of radiation dose between 14 and 300 Gy (J.kg-1). This yield amounted to 0.003 mumol.J-1. The mechanism of formation of this DNA-protein cross-link is thought to result from H atom abstraction by hydroxyl radicals from the methyl group of thymine followed by the addition of the resultant thymine radical to the carbon 3 position of the tyrosine ring and subsequent oxidation of the adduct radical. PMID- 2545258 TI - NMR studies of abasic sites in DNA duplexes: deoxyadenosine stacks into the helix opposite acyclic lesions. AB - Proton and phosphorus NMR studies are reported for two complementary nonanucleotide duplexes containing acyclic abasic sites. The first duplex, d(C-A T-G-A-G-T-A-C).d(G-T-A-C-P-C-A-T-G), contains an acyclic propanyl moiety, P, located opposite a deoxyadenosine at the center of the helix (designated APP 9 mer duplex). The second duplex, d(C-A-T-G-A-G-T-A-C).d(G-T-A-C-E-C-A-T-G), contains a similarly located acyclic ethanyl moiety, E (designated APE 9-mer duplex). The ethanyl moiety is one carbon shorter than the natural carbon phosphodiester backbone of a single nucleotide unit of DNA. The majority of the exchangeable and nonexchangeable base and sugar protons in both the APP 9-mer and APE 9-mer duplexes, including those at the abasic site, have been assigned by recording and analyzing two-dimensional phase-sensitive NOESY data sets in H2O and D2O solution between -5 and 5 degrees C. These spectroscopic observations establish that A5 inserts into the helix opposite the abasic site (P14 and E14) and stacks between the flanking G4.C15 and G6.C13 Watson-Crick base pairs in both the APP 9-mer and APE 9-mer duplexes. The helix is right-handed at and adjacent to the abasic site, and all glycosidic torsion angles are anti in both 9-mer duplexes. Proton NMR parameters for the APP 9-mer and APE 9-mer duplexes are similar to those reported previously for the APF 9-mer duplex (F = furan) in which a cyclic analogue of deoxyribose was embedded in an otherwise identical DNA sequence [Kalnik, M. W., Chang, C. N., Grollman, A. P., & Patel, D. J. (1988) Biochemistry 27, 924-931]. These proton NMR experiments demonstrate that the structures at abasic sites are very similar whether the five-membered ring is open or closed or whether the phosphodiester backbone is shortened by one carbon atom. Phosphorus spectra of the APP 9-mer and APE 9-mer duplexes (5 degrees C) indicate that the backbone conformation is similarly perturbed at three phosphodiester backbone torsion angles. These same torsion angles are also distorted in the APF 9-mer but assume a different conformation than those in the APP 9-mer and APE 9-mer duplexes. PMID- 2545261 TI - Lipid peroxidation of phosphatidylcholine liposomes depressed by the calcium channel blockers nifedipine and verapamil and by the antiarrhythmic-antihypoxic drug stobadine. AB - Nifedipine, verapamil and stobadine were tested and compared with butylated hydroxytoluene (BHT) as possible free radical scavengers inhibiting lipid peroxidation in phosphatidylcholine liposomes. Liposomes were peroxidized by incubation in air at 50 degrees C. Verapamil less than nifedipine less than BHT less than stobadine depressed the lipid peroxidation as detected spectroscopically for conjugate diene and thiobarbituric acid product formation. Verapamil and stobadine were tested as OH radical scavengers in a Fenton-type reaction against spin trap 5,5'-dimethyl-1-pyrroline-N-oxide (DMPO), as detected by ESR spectroscopy. The tested drugs competed with DMPO in trapping OH radicals, with stobadine being more effective than verapamil. ESR spectra of nifedipine in the incubated liposomes revealed that nifedipine could be involved in free radical reactions in the liposomes leading to nifedipine-stable radical(s) which were immobilized in the membrane. The obtained results suggest that some of the beneficial effects of the studied drugs can be mediated in disease by their ability to scavenge free radicals and by their protective effect on lipid peroxidation. PMID- 2545262 TI - Stimulation of CTP: phosphocholine cytidylyltransferase and phosphatidylcholine synthesis by calcium in rat hepatocytes. AB - The effects of Ca2+, ionophore A23187, and vasopressin on CTP:phosphocholine cytidylyltransferase were investigated. Cytidylyltransferase is present in the cytosol and in a membrane-bound form on the microsomes. Digitonin treatment caused release of the cytosolic form rapidly. Addition of 7 mM Ca2+ to hepatocyte medium resulted in a 3-fold decrease in cytidylyltransferase released by digitonin treatment (1.7 +/- 0.1 nmol/min per mg compared to 5.1 +/- 0.2 nmol/min per mg in the control). Verapamil, a calcium channel blocker, partially overcame this effect of Ca2+. Ionophore A23187 and vasopressin both mimicked the effect of Ca2+ and resulted in a decrease in cytidylyltransferase release (2.4 +/- 0.1 nmol/min per mg and 2.5 +/- 0.2 nmol/min per mg, respectively) compared to control (3.4 +/- 0.1 nmol/min per mg). In agreement with the digitonin experiments, incubation with 7 mM Ca2+ resulted in a decrease in cytidylyltransferase in the cytosol (from 4.0 to 1.2 mol/min per mg) and a corresponding increase in the microsomes (from 0.6 to 2.4 nmol/min per mg). Verapamil partially blocked this translocation caused by Ca2+. Ionophore A23187 and vasopressin also caused translocation of the cytidylyltransferase from the cytosol to the microsomes. The addition of Ca2+ also resulted in an increase in PC synthesis. With 7 mM Ca2+ in the medium, the label associated with PC increased to 3.8 +/- 0.1.10(6) dpm/dish from 2.7 +/- 0.1.10(6) dpm/dish after 10 min. PC degradation was also affected, since 7 mM Ca2+ in the medium resulted in an increase in LPC formation both in the cell and the medium. We conclude that high concentrations of calcium in the hepatocyte medium can cause a stimulation of CTP:phosphocholine cytidylyltransferase and PC synthesis in cultured hepatocytes. PMID- 2545263 TI - Rapid modulation of rat adipocyte lipoprotein lipase: effect of calcium, A23187 ionophore, and thrombin. AB - The effect of calcium ions on the lipoprotein lipase (LPL) activity in rat adipocytes has been investigated. Incubation of the cells in the absence of extracellular calcium produced a rapid decline of LPL activity in the cells. The enzyme, however, could be immediately reactivated in less than 3 min by the addition of calcium. The degree of reactivation was proportional to the concentration of extracellular calcium. alpha 1 agonists phenylephrine and methoxamine affected LPL activity only slightly, as did vasopressin and angiotensin II. In contrast, calcium ionophore A23187 elicited a quick and transient enzyme activation which reached its peak 4 min after the addition of the drug. Thrombin (0.1 U/ml) produced the most rapid and intense response. The effect of thrombin was already evident 10 s after its addition, and the enzyme activity almost doubled above the basal level. Extracellular calcium was necessary to achieve thrombin activation. Contrary to previous thought, these data support the conclusion that LPL may undergo rapid activation, and that calcium ions are critically involved in this activation process. Thrombin rapidly raises LPL activity and may be one of its physiological activators in vivo. PMID- 2545264 TI - An improved procedure for the purification of ethanolaminephosphotransferase. Reconstitution of the purified enzyme with lipids. AB - Ethanolaminephosphotransferase (CDPethanolamine:1,2-diacylglycerol ethanolaminephosphotransferase, EC 2.7.8.1) has been purified in active form from rat brain microsomes by a two-step chromatographic procedure. Enzyme preparations characterized by high specific activity and stability were obtained supplementing the solubilization and elution buffers, containing 1% Triton X-100, with 0.01% 2,6-di-tert-butyl-4-methylphenol. The specific activity of the purified enzyme was about 1200-times higher than that of the crude solubilized enzyme. The lipid dependence of ethanolaminephosphotransferase was studied both in the presence of Triton X-100 and in detergent-free enzyme preparations. The activity of the detergent-solubilized ethanolaminephosphotransferase was strongly modified by phospholipids. The kinetic behaviour of the enzyme was also dependent on the lipids contained in the aggregates obtained by removal of the detergent from detergent/lipid/protein suspensions. A regulatory role of phospholipids on the activity of the membrane-bound ethanolaminephosphotransferase is discussed. PMID- 2545265 TI - Effects of nutrient and non-nutrient stimuli on cytosolic pH in cultured insulinoma (HIT-T15) cells. AB - Intracellular pH (pHi) was measured in the insulin-secreting HIT-T15 cell line using the pH-sensitive fluorescent dye, 2',7'-bis(carboxyethyl)-5'(6') carboxyfluorescein (BCECF). It was observed that the addition of a weak acid (e.g., acetate or propionate) caused a rapid decrease in pHi, followed by a slower recovery to the resting pH value. Conversely the addition of N4Cl caused an increase in pHi followed by recovery. The addition of amiloride caused a fall in pHi; however, in this case no recovery to basal pH levels was observed. Subsequent addition of a weak acid caused a further fall in pHi with no recovery. The addition of glucose caused a transient acidification followed by alkalinization. When glucose was added to cells which had been pretreated with amiloride, the initial acidification was not followed by recovery or alkalinization. Addition of glyceraldehyde, alpha-ketoisocaproate, lactate or pyruvate to HIT cells also resulted in intracellular acidification followed by recovery. Similarly, depolarisation of HIT cells by treatment with high K+ or with Ba2+ was associated with a pronounced fall in pHi, followed by a gradual recovery. Insulin secretion from HIT cells was stimulated by glucose, glyceraldehyde, alpha-ketoisocaproate, lactate, pyruvate and KCl, whilst amiloride and weak acids exerted only modest effects in the absence of glucose, but amiloride in particular markedly potentiated glucose-induced insulin release. Thus, HIT cells appear to have an amiloride-sensitive mechanism for the extrusion of protons, probably Na+-H+ exchange. Whilst intracellular acidification appears to potentiate secretory responses to nutrient stimuli, it seems unlikely that the activation of HIT cells by these nutrients occurs as a result of intracellular acidification. The mechanisms by which various nutrient and non-nutrient stimuli might exert distinct effects on pHi are discussed. PMID- 2545266 TI - Inhibition of endothelial secretion of tissue-type plasminogen activator and its rapid inhibitor by agents which increase intracellular cyclic AMP. AB - We investigated the effect of agents which raise intracellular levels of cyclic AMP (cAMP) on the secretion of tissue-type plasminogen activator (t-PA) and type 1 plasminogen activator inhibitor (PAI-1) by cultured human umbilical-vein endothelial cells. Significant inhibition of baseline (unstimulated) t-PA and PAI 1 secretion was observed in response to several agents which, when added exogenously, cause increased intracellular cAMP: cholera toxin, 1-methyl-3 isobutylxanthine (MIX), dibutyryl-cAMP, and prostaglandin E1. These agents also significantly reduced or abolished the previously reported stimulatory effects of thrombin and histamine on t-PA secretion, and, with the exception of MIX, significantly reduced the previously reported stimulatory effect of thrombin on PAI-1 secretion. MIX at a concentration (10 microM) below that required to inhibit t-PA and PAI-1 secretion when tested alone, significantly increased the inhibitory effects of cholera toxin, dibutyryl-cAMP, and prostaglandin E1 on both t-PA and PAI-1 secretion. The data suggest that elevated intracellular levels of cAMP inhibit both spontaneous endothelial secretion of t-PA and PAI-1, and secretion induced by agents (thrombin and histamine) which stimulate endothelial phosphoinositide metabolism, consistent with bidirectional regulation of endothelial fibrinolytic protein secretion by the adenylate cyclase and phosphoinositide signal transduction pathways. The inhibitory effects of cAMP do not appear to be specific for t-PA and PAI-1, since cholera toxin and MIX also inhibited endothelial secretion of the adhesive protein, fibronectin. Significant inhibition of baseline endothelial t-PA and PAI-1 secretion was also caused by the stable prostacyclin analogue iloprost (ZK 36 374) and by arachidonic acid, which is converted by endothelial cells to prostacyclin, suggesting that prostacyclin produced endogenously by endothelial cells may inhibit secretion of fibrinolytic proteins by increasing intracellular cAMP. PMID- 2545267 TI - Differential effects of osmotic pressure on mitochondrial respiratory chain and indices of oxidative phosphorylation. AB - Oxidative phosphorylation was critically evaluated in terms of activities which are sensitive and insensitive to variations in external osmotic pressure in mitochondria. Integrity of mitochondria was determined in terms of a variety of parameters, including the latency of the occluded enzymes, by careful titrations as a function of external osmotic pressure as well as detergent concentrations. The evidence indicated that the rate-limiting step in respiratory states 2 and 4 would be osmotically insensitive, as opposed to the osmotically sensitive respiration of states 1 and 3 and uncoupler-stimulated respiration with glutamate + malate and succinate. Cytochrome oxidase activity in mitochondria as well as in purified reconstituted systems exhibited osmotic insensitivity but marked sensitivity to ionic strength, offering an interesting model to study the osmotically insensitive respiration. Cytochrome oxidase activity led to permeation of mannitol across the mitochondrial inner membrane. Stimulation of cytochrome oxidase activity by uncouplers did not require an intact membrane. PMID- 2545268 TI - Analytical characterization of cytochrome oxidase preparations with regard to metal and phospholipid contents, peptide composition and catalytic activity. AB - A number of preparations of cytochrome oxidase have been analyzed for metals by energy dispersive X-ray fluorescence spectrometry. The EPR characteristics, the peptide compositions, the protein and phospholipid contents as well as the catalytic constants of the samples have also been determined. It is confirmed that the enzyme functional unit contains three copper atoms and one zinc atom in addition to two iron atoms. On the basis of the parameters determined for the different samples it is suggested that a high catalytic activity of a preparation can be correlated to a number of other analytical characteristics. PMID- 2545269 TI - EXAFS studies of the isolated bovine heart Rieske [2Fe-2S]1+(1+,2+) cluster. AB - Recently the involvement of one or, more likely, two nitrogen-ligands in the Rieske-type [2Fe-2S] cluster has been reported based on the chemical assay and various spectroscopic analyses, such as EPR, Mossbauer, ENDOR, and resonance Raman, of isolated Thermus thermophilus HB-8 protein by Fee and his collaborators. Similarly, the presence of at least one nitrogen ligand was shown in the mitochondrial Rieske [2Fe-2S] cluster. We have conducted EXAFS studies of the Rieske [2Fe-2S] protein isolated from the cytochrome bc1 complex of bovine heart mitochondria. Standard analysis could not distinguish one or two nitrogen ligands per cluster. However, one nitrogen and three cysteine ligands per cluster was found to be, possibly, a better solution in more comprehensive analysis procedures. PMID- 2545270 TI - Ethylenediamine as active site probe for Na+/K+-ATPase. AB - (1) Ethylenediamine is an inhibitor of Na+- and K+-activated processes of Na+/K+ ATPase, i.e. the overall Na+/K+-ATPase activity, Na+-activated ATPase and K+ activated phosphatase activity, the Na+-activated phosphorylation and the Na+ free (amino-buffer associated) phosphorylation. (2) The I50 values (I50 is the concentration of inhibitor that half-maximally inhibits) increase with the concentration of the activating cations and the half-maximally activating cation concentrations (Km values) increase with the inhibitor concentration. (3) Ethylenediamine is competitive with Na+ in Na+-activated phosphorylation and with the amino-buffer (triallylamine) in Na+-free phosphorylation. Significant, though probably indirect, effects can also be noted on the affinity for Mg2+ and ATP, but these cannot account for the inhibition. (4) Inhibition parallels the dual protonated or positively charged ethylenediamine concentration (charge distance 3.7 A). (5) Direct investigation of interaction with activating cations (Na+, K+, Mg+, triallylamine) has been made via binding studies. All these cations drive ethylenediamine from the enzyme, but K+ and Mg+ with the highest efficiency and specificity. Ethylenediamine binding is ouabain-insensitive, however. (6) Ethylenediamine neither inhibits the transition to the phosphorylation enzyme conformation, nor does it affect the rate of dephosphorylation. Hence, we provisionally conclude that ethylenediamine inhibits the phosphoryl transfer between the ATP binding and phosphorylation site through occupation of cation activation sites, which are 3-4 A apart. PMID- 2545271 TI - Activity of myelin membrane Na+/K+-ATPase and 5'-nucleotidase in relation to phospholipid acyl profiles, ganglioside composition and phosphoinositides in developing brains of undernourished rats. AB - The relationship between undernutrition-induced alterations in some myelin membrane-bound enzyme activities and phospholipid fatty acid composition of this membrane was ascertained in developing rat brains. Undernutrition was imposed in pregnant dams through gestation and lactation, (last 10 days of gestation, and through lactation) by feed restriction. Experimental groups of animals received 50% of the amount of diet consumed by controls. Pups born to these mother rats were killed at day 7, 14 or 21 of postnatal age. Myelin membrane was isolated from the major regions of the brain, and analysed for phospholipid fatty acid profiles, phosphoinositides and ganglioside species. While there were no diet related differences in the activities of 5'-nucleotidase (EC 3.1.3.5), myelin phospholipids from cerebella and brain stems of experimental rats exhibited lowered proportions of the long-chain polyunsaturated fatty acids, C20:4 (n - 6) and C22:6 (n - 3) concomitant with elevated activities of ouabain-sensitive Na+/K+-ATPase (EC 3.6.1.4). Levels of diphosphoinositide, triphosphoinositide and trisialogangliosides also decreased in myelin from brains of experimental animals. These results suggest a relationship between myelin phospholipid fatty acid profiles as indicators of membrane unsaturation, and the possibility of allosteric modification of Na+/K+-ATPase activity. PMID- 2545272 TI - Interaction between glycophorin and a spin-labeled cholesterol analogue in reconstituted dimyristoylphosphatidylcholine bilayer vesicles. AB - The interaction between glycophorin and a spin-labeled cholesterol analogue has been investigated by EPR spectroscopy. In vesicles which were reconstituted by the freeze and thaw technique, direct evidence was obtained for a reorganisation of the membrane at low protein content (protein/lipid ratio less than 1:300). From the spin exchange interaction we were able to show a protein-induced clustering of the steroid in fluid and in gel state membranes. Tryptic cleavage of the complete N-terminus of glycophorin vanishes the effect. Whereas the removal of the sialic acid residues by neuraminidase digest had no influence on the EPR spectra. The interaction seems to be cholestane spin label specific since it was not observed with an androstane spin-label. PMID- 2545273 TI - Microelectrode studies of amphotericin B on Na+ and K+ conductance in bullfrog cornea. AB - Addition of 10(-5) M amphotericin B to the tear solution of an in vitro preparation of the frog cornea increased the transepithelial conductance, gt, and decreased the apical membrane fractional resistance, f(R0), in the presence or absence of tear Na+ and Cl-. In the presence of tear Na+ and Cl-, amphotericin B increased the short-circuit current, Isc, from 3.9 to 8.8 microA.cm-2 and changed the intracellular potential, V0, from -48.5 to -17.9 mV probably due to a higher increase in the Na+ than in the K+ conductance. In the absence of tear Na+ and Cl , amphotericin B decreased Isc from 5.5 to about 0 microA.cm-2 due to K+ (and possibly Na+) flux from cell to tear and changed V0 from -35.4 to -63.6 mV due to the increase in conductance of both ions. Increase in the tear K+ from 4 to 79 mM (in exchange for choline), in the presence of amphotericin B and absence of tear Na+ and Cl-, decreased f(R0) from 0.09 to 0.06, increased gt from 0.23 to 0.31 mS, increased Isc from 0.63 to 7.3 microA.cm-2, and changed V0 from -65.5 to 17.3 mV due to the change in EK in the presence of a high conductance in the tear membrane. Similar effects were observed with an increase of tear Na+. Results support the concept that the Na+ conductance opened by amphotericin B in the apical membrane is greater than the K+ conductance. Previously observed transepithelial effects of the ionophore may be explained mostly on the basis of its effect on the apical membrane. PMID- 2545274 TI - beta-Casomorphins alter the intestinal accumulation of L-leucine. AB - Everted sacs of the rat jejunum change the accumulation of [3H]leucine when beta casomorphins (BCMs) or synthetic analogs, in a concentration range of 10(-8) mol/l, are coincubated with the amino acid. BCM5 (BCM fragment 1-5, Tyr-Pro-Phe Pro-Gly) and [D-Ala2]-BCM5-NH2 (Tyr-D-Ala-Phe-Pro-Gly) increase, whereas [D-Pro4] BCM5 (Tyr-Pro-Phe-D-Pro-Gly) decreases the leucine accumulation and [Arg8] vasopressin has no effect. No effect of BCM5 could be observed on the accumulation of the space marker [14C]inulin. Specific binding sites for casomorphins were detected microautoradiographically, exclusively at the epithelial cell layer using [3H][D-Pro4]-BCM5 in competition studies as a model. HPLC analysis revealed that under the experimental conditions about 50% of the studied [D-Pro4]-BCM5 was enzymatically degraded and no intact peptide is accumulated within the samples of everted sacs. From the results we postulate a brush-border receptor contact of the BCMs which induces an alteration of the amino acid uptake. A contraluminal binding of the chemical signals is not likely, because there is no evidence for a transepithelial transport of intact BCMs. The observed effects of the BCMs demonstrate as yet unknown peptide-receptor interactions, probably at the brush-border membrane, with subsequent effects on the nutrient supply. Furthermore, the results support the general hypothesis of distinct peptide-receptor interactions in those types of epithelia in which the cells are connected by tight junctions. PMID- 2545275 TI - [Ion conductance channels of bilayer from sarcoma-45 phospholipids]. AB - Properties of black bilayer membranes formed from sarcoma-45 phospholipids differed from those of normal cell phospholipids bilayers. Two types of Ca2+ channels were discovered: those multiple to 35 pA and those multiple to 90 pA. The study of bilayers from phospholipids of nuclei, mitochondria, microsomal fractions showed that Ca2+-channels were localized in the microsomal fraction of transformed cells. The relationship between DB18C6 diacetylderivatives membrane activity and their effect to growth of sarcoma-45 in vivo was discovered: the Ca2+-channel blocker diacetyl-DB18C6 inhibited the transformed cells growth. PMID- 2545276 TI - [N,N'-dicyclohexylcarbodiimide-sensitive K+-dependent H+ secretion from anaerobically grown E. coli cells]. AB - Dependence of N,N'-dicyclohexylcarbodiimide (DCC)-sensitive H+ secretion of K+ activity was discovered. This dependence took place only in anaerobically grown bacteria, and only at the structural intact DCC-sensitive H+-ATPase complex and K+-ionophore Trk. PMID- 2545277 TI - [Lipoxygenase activation during the development of adaptation reactions in the body]. AB - Lipoxygenase activation was studied in the liver and subcutaneous connective tissue of starving mice by ESR technique. The data obtained were discussed from the view-point of possible participation of lipoxygenase activation in organism adaptation reactions. PMID- 2545278 TI - Comparative studies between the glucose-induced phosphorylation signal and the heat shock response in mutants of Saccharomyces cerevisiae. AB - Addition of glucose to derepressed yeast cells, as well as a heat shock treatment, trigger the phosphorylation of trehalase and of trehalose-6-phosphate synthase. In the present paper, evidence is provided for the requirement of the RAS protein in the transduction of the glucose signal. On the other hand, a heat shock at 52 degrees C for 2 min was able to produce a significant phosphorylating effect even in mutant strains deficient in the GTP binding protein. Moreover, it was shown that this treatment does not affect exclusively the cAMP-dependent protein kinase. The use of a series of mutant strains confirmed that low levels of cAMP favor thermotolerance; the role of trehalose in yeast viability is also discussed. PMID- 2545279 TI - Cardiac arrhythmias modelled by Cai-inactivated Ca2+ channels. AB - A model of cardiac cells incorporating the membrane potential and the intracellular calcium concentration as the two dynamical variables is developed. This model is applied to simple systems of cells to investigate its behavior as a function of the model parameters. Rational entrainment is observed in systems of two coupled pacemaker cells. The propagation of the membrane potential and intracellular calcium concentration through a sheet is simulated. Behavior suggestive of circus movement tachycardias is observed. PMID- 2545280 TI - [Inorganic pyrophosphatase of rat liver cytosol. Isolation and properties]. AB - An electrophoretically homogeneous preparation of rat liver cytosolic pyrophosphatase was obtained by a combination of chromatographic methods. The enzyme molecule is made up of two identical subunits, each about 38 kD. The enzyme is activated by Mg2+ and strongly inhibited by Ca2+. Both cations are bound on a time scale of minutes. Ca2+ binding occurs in two steps. EGTA-like metal chelators activate pyrophosphatase, presumably due to the removal of trace amounts of Ca2+ from solutions. PMID- 2545281 TI - [Cell nuclear DNases: multiplicity and heterogeneity]. AB - In order to determine the ratio of activities of major endonucleases of rat liver chromatin, a stepwise fractionation of cell nuclear extracts by chromatography on phosphocellulose and gel filtration through Toyopearl HW60 was carried out. This procedure resulted in partially purified preparations of Ca2+,Mg2+-dependent endonuclease (55 +/- 10 kD), Ca2+,Mg2+-dependent endonuclease (30 +/- 10 kD), Mn2+-dependent endonuclease (30 +/- 5 kD) and acid cation-independent endonuclease. The Ca2+,Mg2+-dependent endonuclease with Mr of 55 +/- 10 kD made up to 57% of the nuclear extract activity in the presence of Ca2+ + Mg2+ and revealed a high calcium-magnesium synergism. Under the same experimental conditions, the 30 +/- 10 kD enzyme made up to 33% of the nuclear extract activity and revealed a low synergism. The activity of Mn2+-dependent endonuclease made up to 26% of the total nuclear extract activity in the presence of Mn2+, that of acid endonuclease--11% of the extract activity in 1 mM EDTA at pH 5.0. It was assumed that the low molecular weight Ca2+,Mg2+-dependent endonuclease represents a product of limited proteolysis of high molecular weight Ca2+,Mg2+-dependent endonuclease. PMID- 2545282 TI - [Role of cAMP in the regulation of adrenal mineralocorticoid function by potassium ions]. AB - The role of cAMP in K+-dependent regulation of aldosterone output was studied. Guinea pig adrenal slices were incubated with various concentrations of K+ for 20 min. Aldosterone and cAMP output, protein synthesis and phosphorylation were increased with a rise in K+ concentration. The increase in the phosphorylation of the protein with apparent molecular weight of about 38 kD resulted from the increase in K+ concentration in the incubation medium. Proteins from adrenal slices preincubated with 3.0 and 11.0 mM K+ were separated by SDS polyacrylamide gel electrophoresis. No qualitative differences were revealed in autoradiograms of [14C]-prelabeled proteins; however, quantitative differences were present. These results suggest that the cAMP messenger system may be involved in the K+ dependent control of aldosterone biosynthesis. PMID- 2545283 TI - [Dissociation of hexamers of cytochrome P-450 LM2 in the presence of the non ionic detergent emulgin 913]. AB - It was shown that the maximal degree of dissociation of cytochrome P-450 LM2 hexamers in the presence of the nonionic detergent Emulgene 913 (20 degrees C) is observed at the detergent concentration of about 0.2%. Using equilibrium centrifugation in solutions of different density, the molecular mass of the dissociation product minus that of the bound detergent was found to be equal to 50 +/- 8 kDa, thus corresponding to the molecular mass of the monomer. One cytochrome P-450 LM 2 molecule binds 80 +/- 20 molecules of Emulgene 913. PMID- 2545284 TI - [An experimental approach to research on the integrative properties of a single brain neuron]. AB - The difficulties in investigation of the single neuron integrative properties in mammalian brain are connected with the absence of a mean of determination of neuronal set contribution in their activity. In given paper an experimental approach to the decision of this task is proposed. It consists in neuron activity investigation in the model learning training situation under condition of partial functional isolation from nervous cells by means of decrease in extracellular calcium concentration level using ethylene glycol tetraacetate. As indicator of such isolation was a partial or total reduction of responses in investigated neurons to electric microstimulation of neighbouring (200-240 mkm) parts of neocortex. The results of analysis of neuron responses in sensomotor cortex in the rat's brain in the process of acetylcholine repetitive local application give possibility to propose that some neurons don't exhibit plasticity according to the indicator in the dynamics of impulse discharge rate in responses to transmitter. PMID- 2545285 TI - [Second messengers in the regulation of nerve cell plasticity during learning]. AB - A review of 1972 to 1988 publications is presented, as well the data obtained by the authors on the influence of second messengers and their intracellular receptors on neuron plasticity in the learning experimental models. The emphasis is laid on cyclic 3',5'-adenosine monophosphate; cyclic 3',5'-guanosine monophosphate, calcium ions and their intracellular receptor calmodulin, as well as on the metabolites of phosphoinositide exchange: inositol-1, 4, 5 trisphosphate, 1,2-diacylglycerol and protein kinase C activated by the latter and arachidonic acid. PMID- 2545286 TI - Sensitivity of the renal adenylate cyclase receptor system to parathormone during the last third of pregnancy in the rat. AB - During the last third of pregnancy, the rat phospho-calcic metabolism is clearly modified. Sensitivity of the renal receptor to parathormone (PTH) was investigated over this period in the fetuses and their mothers. As early as day 16.5 of pregnancy, the fetal kidney is sensitive to bovine PTH (1-34) on the basis of cAMP production. Sensitivity declines on the last day of pregnancy. The newborn kidney showed a higher response to PTH 3 h after birth than after 24 h. The basal adenylate cyclase activity in renal membranes of maternal kidney does not change. The sensitivity to PTH and to fluoride both decreased on day 18.5. A desensitization process of the PTH receptor A-C system has to be studied in order to explain these results. PMID- 2545287 TI - Post-traumatic stress disorder, hormones, and memory. PMID- 2545288 TI - CSF prostaglandin-E in agoraphobia with panic attacks. AB - Prostaglandins are thought to act as neuromodulators of both central catecholamine and endocrine systems. Abnormalities of these systems have been described in affective disorders, in general, and in agoraphobia with panic attacks, in particular. This study measured basal prostaglandin-E (PGE) cerebrospinal fluid (CSF) levels in 20 patients with agoraphobia with panic attacks and 10 nonpsychiatric controls. In a subgroup of patients and controls, CSF levels of adrenocorticotrophic hormone (ACTH) and corticotropin-releasing factor (CRF) were also measured. There was no significant difference in CSF PGE levels between patients and controls. However, patients with higher depression scores had lower CSF PGE levels. CSF PGE levels tended to correlate with CSF ACTH, but not CSF CRF in the patient group, in general, and in the female patients, in particular. These findings do not support an abnormality in basal CNS PGE production in agoraphobia with panic attacks, but suggest further study of the PGE modulatory effect on the hypothalamic-pituitary-adrenal axis in this disorder. PMID- 2545289 TI - Directionality of neural signals in central nervous system neural networks. AB - These studies are concerned with neuronal population theory of brain function. Neural networks the cerebral cortex and the limbic systems are investigated. We use coherence, partial coherence, phase and cycle time spectral analysis in these studies. The development of software for phase filtering has permitted us to analyze directionality of neural signal flow following coherence and partial coherence estimates. These estimates give an indication of the interaction between the brain regions studied. These data give further insights of the input and output status and the linear aspects of these interactions. Spontaneous and evoked potential analog data are analyzed to evaluate the physiological parameters of these interrelations. PMID- 2545290 TI - Stochastic variables responsible for observed saccadic variability. AB - A new stochastic local feedback model of the horizontal saccadic system based on time optimal neural control within the superior colliculus has been previously described. This model uses a premotor neural circuit composed of burst, tonic, and pause cells and innervates a fourth order linear homeomorphic muscle plant. A sequence of saccades recorded from human subjects shows great variability in peak velocity, final position, and amount and type of post-saccadic behavior. This variability is duplicated in a sequence of simulated saccades through the use of random variables within the neural circuitry. The random variables are agonist burst cell magnitude, antagonist post inhibitory rebound burst magnitude and timing, and to a lesser extent muscle saturation magnitude. These four random variables are shown to cause all the observed variability in human saccades, including: trajectory profile, velocity profile, dynamic overshoot, and glissadic overshoot and undershoot. PMID- 2545291 TI - Identification of multifiber neural spike trains by interval prediction and morphology. AB - Interpretation of neurograms from multifiber preparations is usually limited to forming a grand running average. In so doing, individual nerve activity is lost. This information is important in studying nervous system activity. We have developed a method to find the rates of constituent nerve fibers using linear one step prediction and cross correlation between the spike template and instantaneous spikes. Some size discrimination is employed, as is a memory to mark spikes which are "accounted for." Thus we are able to parse out the several rates which make up an aggregate spike train. Our results show that some afferent baroreceptor fibers may increase rate, while others may decrease rate, in response to holding carotid sinus pressure at a high level for 10 minutes. PMID- 2545292 TI - Teratogenesis of polychlorocycloalkane insecticides in chicken embryos resulting from their interactions at the convulsant recognition sites of the GABA (pro)receptor complex. PMID- 2545293 TI - Tissue mineral element content in swine fed clinoptilolite. PMID- 2545294 TI - Effect of fenvalerate on metabolic ion dynamics in the fathead minnow (Pimephales promelas) and bluegill (Lepomis macrochirus). PMID- 2545296 TI - Effect of heavy metals ions on enzyme activity in the Mediterranean mussel, Donax trunculus. PMID- 2545295 TI - Gill ATPase activity in Procambarus clarkii as an indicator of heavy metal pollution. PMID- 2545299 TI - Specific L-arginine taste receptor sites in the catfish, Ictalurus punctatus: biochemical and neurophysiological characterization. AB - We report here the characterization of the arginine binding site(s) and corroborative neurophysiological studies. Binding of L-[3H]arginine to Fraction P2 from taste epithelium was measured by a modification of the method of Krueger and Cagan. Parameters for measuring maximal binding activity were established for both duration of incubation and pH of medium. At pH 7.8, the apparent single rate constant for association (kobs) at 4 degrees C was 4.72 x 10(+5).M-1.min-1. Dissociation was more complex, yielding two rate constants of 1.77.min-1 and 8.34 x 10(-3).min-1. These data suggest the presence of two affinity states for L arginine. The KD values as calculated from the ratio k-1/k+1 were 1.3 x 10(-6) M and 1.8 x 10(-8) M. Homologous inhibition studies of L-arginine binding were not fit by a simple mass action relationship (Hill Coefficient 0.79), but were best fit by a two-site model with IC50 values of 1.6 x 10(-6) M for the high affinity state and 9 x 10(-4) M for the low affinity state. Multiunit neural recordings examined the stimulatory effectiveness of a number of guanidinium-containing compounds. Compared with L-arginine, only L-arginine methyl ester and L-alpha amino-beta-guanidino propionic acid (L-AGPA) were effective stimuli. Cross adaptation experiments demonstrated that at 10(-4) M L-arginine methyl ester, L AGPA and, to a lesser extent, D-arginine were effective cross-adapting stimuli to 10(-6) M L-arginine. In competition binding studies L-arginine methyl ester, L AGPA and D-arginine also inhibited binding of L-[3H]arginine (10(-6) M), but each recognized only one affinity state. Inhibition by the poorly cross-adapting stimuli L-glutamate, glycine and L-alanine occurred only above 10(-3) M, indicating that the binding sites for L-arginine are selective. These studies suggest that there are at least two affinity states of L-arginine binding, that the binding sites are specific, and that effective agonists of L-arginine receptors must contain a guanidinium group and an unblocked L-alpha-amino group. PMID- 2545297 TI - Elevation of intracellular cyclic AMP concentration fails to inhibit adrenaline induced hyperpolarization in amphibian sympathetic neurons. AB - 1. The effect of drugs on the adenosine 3':5'-cyclic monophosphate (cyclic AMP) content of desmethylimipramine (DMI)-treated bullfrog paravertebral sympathetic ganglia was studied by radioimmunoassay. The adrenaline-induced hyperpolarization (Adh) in the tissue was recorded by means of the sucrose-gap technique. 2. In the presence of propranolol (1 microM) and DMI (0.5 microM), adrenaline (1 microM) significantly reduced the concentration of cyclic AMP in forskolin-treated ganglia. This effect was prevented by pertussis toxin (5 micrograms ml-1). 3. The relative potency for drugs which increased ganglionic cyclic AMP content was: 50 microM forskolin much greater than 5 mM fluoride greater than 2 mM fluoride greater than 2 mM isobutylmethylxanthine (IBMX) greater than 5 mM caffeine. In contrast, their relative potency for inhibition of the Adh was: 2 mM IBMX greater than 5 mM fluoride greater than 5 mM caffeine much greater than 2 mM fluoride greater than 50 microM forskolin. The Adh was unaffected by pertussis toxin (5 micrograms ml-1). 4. Although the Adh was slightly reduced by the extracellular application of 8-bromo (8-Br) cyclic AMP, the majority of the data suggest that the transduction mechanism underlying the Adh is independent of the intracellular cyclic AMP concentration and provide an example of an alpha 2-adrenoceptor mediated response that occurs independently of inhibition of adenylate cyclase. PMID- 2545300 TI - Activation of mu opioid receptors in the nucleus raphe dorsalis blocks apomorphine-induced aggression in rats: serotonin appears not to be involved. AB - The role of mu opioid receptors in the nucleus raphe dorsalis (DR) in the control of apomorphine-induced aggression was studied in rats. Administration in the DR of a selective mu opioid receptor agonist, (D-Ala2,N-Me-Phe4,Gly5-ol)-enkephalin (DAGO), in doses ranging from 0.01 to 1 microgram/0.5 microliter, dose dependently reduced aggression caused by apomorphine 20 mg/kg intraperitoneally. 0.01 microgram DAGO significantly reduced the time spent by the animals in aggressive posture and 0.1 and 1 microgram markedly reduced both aggressive postures and attacks. 5 micrograms (in 0.5 microliter) naloxone in the DR completely antagonized the anti-aggressive effect of DAGO (0.1 microgram/0.5 microliter) injected in the same area. 0.1 and 1 microgram but not 0.01 microgram DAGO significantly increased serotonin (5-HT) metabolism in the striatum, a terminal area almost exclusively innervated by DR, indicating that the activity of 5-HT cells in the DR was modified by DAGO. In animals given 6 micrograms/3 microliters 5,7-dihydroxytryptamine in the DR 11 days before, in which striatal 5 HT levels were markedly depleted, no significant changes were found in the time spent by the apomorphine-treated animals in aggressive postures, numbers of attacks or anti-aggressive effect of 0.1 and 1 microgram DAGO administered in the DR. The study shows for the first time that activation of mu opioid receptors in the DR has a powerful anti-aggressive effect in one model of experimental aggression by a mechanism apparently not involving changes in the activity of 5 HT cells in this area. PMID- 2545298 TI - Kinins act on B1 or B2 receptors to release conjointly endothelium-derived relaxing factor and prostacyclin from bovine aortic endothelial cells. AB - 1. Bradykinin (Bk) induced the coupled release of endothelium-derived relaxing factor (EDRF) and prostacyclin (PGI2) from bovine aortic endothelial cells grown in culture. The B2 kinin receptor antagonist, [D-Arg0,Hyp3,Thi5,8,D-Phe7]-Bk, abolished this release by Bk. 2. Des-Arg9-Bk, a B1 kinin receptor agonist, also induced the release of EDRF and PGI2, but much higher concentrations were required to obtain a similar release to that induced by Bk. 3. [Leu8],des-Arg9 Bk, a B1 receptor antagonist, significantly reduced the response to des-Arg9-Bk without affecting the release induced by Bk. 4. The release of EDRF and PGI2 induced by arachidonic acid or ADP was not significantly affected by the B2 or the B1 antagonist. 5. We conclude, therefore, that Bk and des-Arg9-Bk were acting respectively on B2 and B1 bradykinin receptors. 6. The possible role of kinin receptors in the release of EDRF and PGI2 from endothelial cells is discussed. PMID- 2545302 TI - Facilitation by forskolin of electrically evoked vasopressin release in vitro requires bursting pattern of stimulation. AB - Electrically stimulated release of vasopressin from isolated rat neurohypophysis was measured in the presence and absence of forskolin. Forskolin inhibited the release of vasopressin evoked by continuous stimulation at constant frequency, and enhanced the release evoked by trains of pulses separated by silent intervals. It is suggested that this biphasic effect of forskolin is consistent with an involvement of cyclic adenosine monophosphate (cAMP) in the regulation of cytosolic Ca2+ in the neurosecretory terminals. These results may partly explain some of the discrepant effects of increasing tissue cAMP observed between those studies using electrical pulses, and those using high K+, for stimulation of vasopressin release. PMID- 2545301 TI - Characterization of the relation between sodium channels and electrical activity in cultured rat skeletal myotubes: regulatory aspects. AB - The relation among sodium channel density, frequency of electrical activity and maximal rate of rise of the action potential was studied in developing and mature rat skeletal myotubes in culture. The number of tetrodotoxin (TTX)-sensitive Na channels was determined by measurements of the amount of [3H]saxitoxin (STX) bound to the cultures, and electrical properties were recorded with intracellular microelectrodes. The EC50 for TTX-induced decreases in maximal STX-binding, frequency and rate of rise of action potentials was in the range 8-20 nM. The 3 variables increased in parallel with age in culture to reach peak values at age 7 8 days, and then decreased in parallel until 10-12 days in culture. The age related increase in Na-channel density was decreased, but not abolished, by prevention of myoblast fusion. Treatment with the Ca2+ ionophore, A23187, down regulated, and blockade of Ca-channels with verapamil up-regulated the number of Na-channels. Na-channel density was also increased by chronic treatment with TTX and elevated external [K+], which eliminated spontaneous electrical and contractile activity. Parallel effects were observed on frequency and rate of rise of action potentials. Up-regulation of Na-channels was prevented by simultaneous treatment of myotubes with inhibitors of protein synthesis. We conclude that electrical and mechanical activity of cultured myotubes regulate de novo synthesis of Na-channels through alterations in the level of cytosolic Ca2+. PMID- 2545303 TI - Autoradiographic localization of mu-opioid and neurotensin receptors within the mesolimbic dopamine system. AB - In vitro autoradiographic techniques were coupled with selective chemical lesions of the A10 dopamine cells and intrinsic perikarya of the A10 region to delineate anatomical localization of the mu-opioid receptors, labeled with 125I-Tyr-D-Ala NMePhe-Gly-OH (125I-DAGO), and neurotensin receptors, labeled with 125I [Tyr3]neurotensin within the mesolimbic dopamine system. Unilateral lesions of dopamine perikarya produced by 6-hydroxydopamine (6-OHDA), administered in the ventral mesencephalon, produced a unilateral loss of specific neurotensin binding (65%), but did not affect mu-opioid receptor density or distribution. Unilateral lesions of intrinsic perikarya by quinolinic acid (250 nmol) injected into the A10 dopamine region produced a significant reduction in mu-opioid receptors (50%), as well as a concomitant reduction in neurotensin receptors. Unilateral 6 OHDA- or quinolinic-induced lesions in the ventral mesencephalon failed to cause significant reduction in mu-opioid receptors in the caudate putamen or limbic forebrain. In contrast, mesencephalic lesions produced significant reductions (50%) in neurotensin binding in the caudate putamen and lateral nucleus accumbens. However, neurotensin binding within the medial nucleus accumbens and adjacent limbic nuclei were unaffected by these treatments. These results are consistent with the pharmacological effects of mu-opioids and neurotensin, and suggest an indirect modulation of the mesolimbic dopamine neurons by mu-opioid agonists and the endogenous opioid peptides. PMID- 2545304 TI - Functional substance P receptors on a human astrocytoma cell line (U-373 MG). AB - [125I]Bolton Hunter conjugate of substance P ([125I]BHSP) can bind to human astrocytoma membranes in a monophasic and saturable manner with a Kd of 0.57 +/- 0.17 nM and a Bmax of 67.8 +/- 5.5 fmol/mg protein. The rank order of potency of tachykinins and related analogues as inhibitors of [125I]BHSP binding to astrocytoma membranes and intact cells correlated with their relative abilities to stimulate uridine incorporation into nucleic acid. The observed specificity pattern conformed to that reported for the NK1 tachykinin receptor with SP much greater than eledoisin greater than neurokinin A greater than neurokinin B and [Glp6, L-Pro9]SP(6-11) much greater than [Glp6, D-Pro9]SP(6-11). PMID- 2545305 TI - The distribution of neurotensin receptors and acetylcholinesterase in the human caudate nucleus: evidence for the existence of a third neurochemical compartment. AB - The distribution of neurotensin receptors in the human caudate nucleus was studied using autoradiographic methods following in vitro labelling of cryostat sections with [3H]neurotensin, and the pattern of receptor labelling was compared to the distribution of acetylcholinesterase (AChE) staining in adjacent sections. A heterogeneous pattern of neurotensin receptors was found in the caudate nucleus. Patches of low receptor density aligned with the AChE-poor striosomes, regions of moderate receptor density corresponded with the AChE-rich matrix zone, and annular regions of high receptor density aligned with the AChE-negative border zone lying between the AChE-poor striosome and the AChE-rich matrix compartments. These results suggest the existence of 3 neurochemical compartments within the human caudate nucleus. PMID- 2545306 TI - Chronic septal lesions cause upregulation of cholinergic but not noradrenergic hippocampal phosphoinositide hydrolysis. AB - The functional integrity of hippocampal muscarinic and noradrenergic receptors was assessed utilizing phosphatidyl inositol (PI) hydrolysis. Four months following septal lesions an enhanced hippocampal PI response to carbachol stimulation (61%) was observed while no effect was found in maximal noradrenergic stimulation. PMID- 2545308 TI - Forskolin reduces a transient potassium current in lobster neurons by a cAMP independent mechanism. AB - Forskolin decreases the transient potassium current, IA, in voltage-clamped somata of identified neurons in the stomatogastric ganglion of the spiny lobster, Panulirus interruptus. The diterpene reduces the peak outward current and accelerates the rate of inactivation of IA. Forskolin has no detectable effects on two other identifiable potassium currents in these cells, IK(Ca) and IK(V). Three identified stomatogastric neuron types (PD, PY, AB) have marked amounts of IA which are affected by forskolin; three other cell types (LP, IC, VD) have little or no IA, and forskolin has no effect on their outward currents. Bath application of 8-bromo-cAMP, N,N-dibutyryl-cAMP and IBMX do not affect IA. In addition, the forskolin analog, 1,9-dideoxyforskolin, which does not activate adenylate cyclase, mimics forskolin's effects on IA. Thus, the effects of forskolin on IA are not mediated by cAMP elevation. PMID- 2545307 TI - Novel recognition site for L-quisqualate sensitizes neurons to depolarization by L-2-amino-4-phosphonobutanoate (L-AP4). AB - Brief exposure of rat hippocampal slices to L-quisqualate sensitizes pyramidal neurons to depolarization by L-2-amino-4-phosphonobutanoate (L-AP4). We report here experiments designed to clarify the duration, pharmacology, mechanism, and pathway specificity of this 'QUIS-effect'. The quisqualate-induced sensitization to L-AP4 decreases only 3-fold over a 4 h period. No compound besides quisqualate has been found to induce the QUIS-effect, including quisqualate analogues, potent excitatory amino acid agonists, L-glutamate, L-aspartate, and compounds known to stimulate second messenger systems in hippocampal slices. Of 43 compounds assayed here, only 5 are able to block the induction of the QUIS-effect. Although these blockers are also potent ligands at a chloride-dependent glutamate uptake site, the marked difference in rank ordering of compounds for QUIS-effect blockade and uptake site potency suggests that the QUIS-effect is not induced through this uptake site. The QUIS-effect can be induced in the CA1 region, the medial perforant path, and the lateral olfactory tract of the rat, and in the guinea pig CA1. It cannot be induced in the L-AP4-sensitive rat lateral perforant path (LPP), suggesting that the receptors for L-AP4 in the LPP may be distinct from those that are sensitized by quisqualate in the other pathways. PMID- 2545309 TI - Chronic ethanol treatment alters the behavioral effects of Ro 15-4513, a partially negative ligand for benzodiazepine binding sites. AB - Pentylenetetrazol (PTZ)-induced convulsion were studied in control, chronic ethanol-maintained, and ethanol-withdrawal rats. The convulsive doses of PTZ varied among the different groups of rats. Ethanol-maintained rats required higher doses of PTZ to produce convulsions, compared to control and ethanol withdrawal rats. The partially negative ligands for benzodiazepine binding sites, Ro 15-4513 (2 mg/kg, i.p.) and FG 7142 (20 mg/kg, i.p.) produced proconvulsant effect in saline (control) and ethanol-withdrawal rats as they potentiated the effect of subconvulsive dose of PTZ. A higher dose of Ro 15-4513 (4 mg/kg, i.p.), but not FG 7142 (up to 80 mg/kg, i.p.), also produced proconvulsant effect in ethanol-maintained rats. Furthermore, Ro 15-4513 (5, 10 mg/kg, i.p.), but not FG 7142 (up to 80 mg/kg, i.p.), produced clonic-tonic seizures of short duration in ethanol-withdrawal rats. These effects of Ro 15-4513 and FG 7142 were reversed by diazepam (2 mg/kg, i.p.), as well as by the GABA-neutral Ro 15-1788 (10 mg/kg, i.p.), thereby, indicating the involvement of central benzodiazepine receptors in the action of Ro 15-4513 and FG 7142. These observations suggest that chronic ethanol treatment selectively alters the receptor sensitivity to Ro 15-4513, an ethanol antagonist and partially negative ligand for BZ sites, and this observation supports the notion that ethanol effects are more susceptible to reversal by the imidazobenzodiazepine as compared to other negative ligand for BZ binding sites. PMID- 2545310 TI - Delayed effects of chronic cortisol treatment on brain and plasma concentrations of corticotropin (ACTH) and beta-endorphin. AB - The effects of 2-day and 7-day cortisol treatment on immunoreactive corticotropin (ACTH) and beta-endorphin concentrations were measured in the cerebral cortex, hippocampus, hypothalamus, and cerebellum in male rats. Plasma ACTH, beta endorphin, corticosterone, and cortisol levels were also measured in parallel. Cortisol administration by osmotic minipumps (25 mg/kg/day) maintained a constant, moderately high concentration (23.0 +/- 2.7 micrograms/100 ml) of this glucocorticoid in plasma. Two-day cortisol treatment suppressed the plasma concentration of ACTH and corticosterone, and also decreased, to a lesser degree, concentrations of beta-endorphin. ACTH and beta-endorphin levels in the brain remained unchanged after 2 days of cortisol treatment. After 7-day treatment, however, plasma concentrations of ACTH and beta-endorphin further decreased, while ACTH and beta-endorphin concentrations in the cortex and beta-endorphin concentrations in the cerebellum were also significantly decreased. Peptide concentrations in other brain areas did not change significantly with either 2 day or 7-day cortisol treatment. These data suggest that there are delayed effects of glucocorticoids on pro-opiomelanocortin peptide secretion and/or metabolism in the central nervous system. These findings are consistent with the impaired cognitive functions of patients with diseases, such as Cushing's syndrome and depression, that have long-lasting elevated cortisol secretion. PMID- 2545311 TI - Characterization of polyclonal antibodies to brain protein phosphatase 2A and immunohistochemical localization of the enzyme in rat brain. AB - Polyclonal antibodies to the catalytic subunit of protein phosphatase 2A (PrP-2A) from bovine brain were prepared by immunizing rabbits and then purified by antigen-affinity column chromatography. The purified antibodies recognized only PrP-2A among proteins examined, including calcineurin and PrP-1. The antibodies cross-reacted only with a protein in the crude homogenate from rat brain, which comigrated with the catalytic subunit of PrP-2A on SDS-PAGE. The antibodies completely inhibited the activity of PrP-2A, and immunoprecipitated the purified enzyme. Immunoblot analysis demonstrates that, among the subcellular fractions from rat brain, the cytosol fraction and the synaptosomal cytosol fraction show high immunoreactivities, and that any of examined regions of rat brain shows immunoreactivity more or less, in which the caudatoputamen was highest. Immunohistochemical studies demonstrate that the enzyme is distributed widely in various regions of rat brain and that the immunoreactivity is localized mainly in neurons. In general, immunostaining of neurons was strong in neurites as well as somata. It was noted that intracerebellar nuclei were strongly stained in both neuronal somata and dendrites. Amygdaloid complex, thalamus, neocortex, hippocampal formation, and caudatoputamen were moderately stained. PMID- 2545312 TI - Stimulation of dopamine release in the rat neostriatum in vivo by activation of the voltage-sensitive sodium channel by scorpion venom neurotoxin. AB - Scorpion venom neurotoxins open sodium channels and thus may enhance neurotransmitter release by increasing membrane permeability to sodium. This study carried out in vivo examined the effects of the scorpion Androctonus australis neurotoxin (ScAaTx) on the levels of dopamine (DA) in push-pull perfusates of the striatum of chloral hydrate-anaesthetised rats. ScAaTx (2.5, 5.0 and 10.0 ng/microliters) stimulated DA release in a dose-dependent manner. The release of DA induced by ScAaTx (10 ng/microliters) was completely blocked when the brain site was perfused with Ca2+-free CSF containing 2 mM EGTA or in the presence of TTX (10(-5) M). These results indicate that the potent stimulatory effects of ScAaTx on neostriatal DA release in vivo are mediated via voltage-sensitive sodium channels. PMID- 2545313 TI - Effect of feeding clinoptilolite (zeolite) to three strains of laying hens. AB - 1. One hundred and twenty 16-week-old, single combed pullets of three strains were fed on a diet containing 160 g protein/kg with or without 50 g clinoptilolite/kg in a trial with 20 hens per treatment. Sterile river sand replaced clinoptilolite in the control diet in order to keep the diets isonergetic. 2. The hens were individually caged in a naturally-ventilated laying house and fed on one of the two diets for ten 28-d periods. 3. No significant dietary effects between treatments were observed with respect to body weight, age at first egg, egg weight, Haugh scores or food intake/hen. 4. Significant dietary effects in favour of clinoptilolite feeding were noticed with the number of eggs laid per hen, shell thickness, efficiency of food utilisation, droppings moisture content and mortality. 5. Significant differences between strains were observed with respect to all measurements taken except food intake/hen d. PMID- 2545314 TI - Adrenocortical and heterophil/lymphocyte responses to challenge in hens showing short or long tonic immobility reactions. AB - 1. Adult White Leghorn hens showing short or long tonic immobility reactions were classified as low-fear (LF) or high-fear (HF) responders, respectively. Following cannulation, their adrenocortical responsiveness to ACTH administration and the effects of chronic frustration induced by thwarting of feeding on plasma corticosterone concentrations and heterophil/lymphocyte (H/L) ratios were measured at regular intervals. 2. ACTH injection elicited significant and similar mean increases in circulating corticosterone concentrations in both the LF and HF groups. 3. Absolute H/L ratios were higher in HF than in LF hens after cannulation, although both groups showed similar proportional increases from pre operation ratios. 4. Plasma corticosterone concentrations and H/L ratios were significantly increased at 20 h and 44 h respectively after the frustration of feeding regime began. Thus, adrenocortical activation preceded increases in H/L ratios but, whereas the elevated plasma corticosterone concentrations found after 20 h remained virtually constant thereafter, H/L ratios rose progressively throughout the 68 h frustration period. 5. There were no significant differences between groups in their plasma corticosterone and H/L responses to frustration. However, a consistent trend towards greater adrenocortical activation was observed in the HF than in the LF hens. PMID- 2545315 TI - Influence of fibre on metabolisable energy of diet and performance of growing pullets in the Tropics. AB - 1. Diets in which maize and palm kernel meals were major ingredients and whose metabolisable energy values were varied by the inclusion of graded amounts of maize cob (50 to 200 g/kg diet) were fed to growing pullets for 9 to 21 weeks of age. 2. Apparent metabolisable energy decreased from 11.82 to 9.79 MJ/kg diet as neutral detergent fibre increased from 182 to 330 g/kg. 3. Metabolisable energy intake was significantly reduced when the maize cob content was above 100 g/kg diet. 4. Protein intake was unaffected by treatment but apparent nitrogen retention was significantly depressed for pullets receiving the diet with 200 g/kg maize cob. Birds on the diet did not come into lay during the study. PMID- 2545317 TI - Experimental studies on Yersinia enterocolitica infection in chickens exposed at 1-day old. AB - 1. Seventy 1-d-old broiler chicks were experimentally inoculated orally with Yersinia enterocolitica serotype 0:3 (1.4 x 10(11) cells/chick), 0:8 (1.6 x 10(11) cells) and 0:9 (8.0 x 10(10) cells) with or without sodium bicarbonate solution (10 g/l). 2. None of the chicks showed any overt clinical signs or pathological lesions although the organism was demonstrated in the ileum and shedding was observed up to 13 d after exposure. 3. The serotype, dose of Y. enterocolitica and administration of NaHCO3 solution had no significant effect on the weight gain of exposed broiler chicks. 4. Y. enterocolitica was isolated from the liver, spleen, heart and gall bladder of infected chicks 70 d after exposure. 5. Although broiler chicks appear resistant to high doses of Y. enterocolitica by the oral route, detection of the organism in the organs of infected chickens is of public health significance. PMID- 2545316 TI - Genetic differences in susceptibility to a mixture of avian infectious bronchitis virus and Escherichia coli. AB - Two-week-old chickens of 9 inbred and partially inbred lines of chickens were challenged intranasally with a mixed infection consisting of a pool of virulent strains of infectious bronchitis virus and a pool of pathogenic strains of Escherichia coli. 2. Wide differences in mortality were observed in the different lines, ranging from 3% in a Brown Leghorn line to 87% in White Leghorn line 7(2). 3. Experiments involving challenge with E. coli alone or virus alone suggested that this variation reflected resistance to the virus rather than to E. coli. 4. Reciprocal F1 matings suggested these differences in mortality were not attributable to maternal effects and indicated that the inheritance of resistance was fully dominant. 5. The pattern of mortality in F2 and backcross progeny of matings was compatible with the inheritance of a dominant autosomal resistance gene and showed no evidence of association with the major histocompatibility complex. PMID- 2545318 TI - Anogenital warts in childhood. AB - Fifteen children with anogenital warts are presented. Twelve cases were referred for assessment of sexual abuse which was established in six cases, strongly suspected in one, and excluded in three. In two, the source was unclear. Papillomavirus typing was carried out by HPV DNA dot and Southern blot hybridization using mixed HPV 6/11, 16/18, and 2/3 DNA probes on 15 specimens from 12 of the children. Seven biopsy specimens were positive for HPV 6 or 11 and one hybridized with both HPV 6/11 and 16/18 mixed sets of probes. Two specimens were positive for HPV 2, and a further two hybridized with both HPV 18 and 2. Three wart specimens could not be typed with the available genital or skin probes. The viruses causing genital tract papillomata are the same for children and adults. The identification of HPV 16/18 raises the concern of potential oncogenicity and stresses the need for long-term assessment. The diagnosis of sexual abuse was made on history rather than examination, as only two cases showed additional physical signs of sexual abuse. It is advocated that the presence of anogenital warts alone be sufficient grounds to pursue the possibility of sexual abuse. Nonsexual transmission, although possible, is far less likely. PMID- 2545320 TI - Lack of cardiac alpha 1-adrenoceptor involvement in ethanol-induced cardiac hypertrophy. AB - The development of cardiac hypertrophy was examined in rats given ethanol in a nutritionally adequate, liquid diet mixture, by intubation, in severely intoxicating doses at 8-h intervals for up to 96 h, alone or in combination with prazosin. Other groups of rats received isocalorically paired quantities of maltose-dextrin. Adrenal glands of rats receiving ethanol were larger than those from control animals. Prazosin did not affect this measure. In contrast, concurrent treatment with prazosin enhanced the loss of medullary catecholamines and noradrenaline from hearts of rats given ethanol, while it had no such effects in controls. Reflecting these changes, excreted quantities of catecholamines were markedly increased in rats given ethanol and prazosin. Hearts of animals given the combined treatment of ethanol and prazosin showed cardiomegaly at 24 h, when there was an increase of about 20% in proportional heart weight, an increase that persisted through the remaining 3 days of the study. At 48 h, hearts of animals given prazosin and ethanol were heavier than those given ethanol alone. A significant correlation between catecholamine excretion and the development of cardiac hypertrophy was identified. The results of the study show that prazosin can enhance effects of ethanol on the peripheral sympathetic nervous system. Moreover, the results suggest that postsynaptic alpha 1-adrenoceptor stimulation in the heart is not an important contributor to ethanol-induced cardiomegaly. PMID- 2545321 TI - Thermal and water source effects upon the stability of enteroviruses in surface freshwaters. AB - The long-term survival of three human enterovirus serotypes, Coxsackievirus B3, echovirus 7, and poliovirus 1 was examined in samples of surface freshwater collected from five sites of physically different character. These were an artificial lake created by damming a creek, a small groundwater outlet pond, both a large- and a medium-sized river, and a small suburban creek. Survival was studied at temperatures of -20, 1, and 22 degrees C. The average amount of viral inactivation was 6.5-7.0 log10 units over 8 weeks at 22 degrees C, 4-5 log10 units over 12 weeks at 1 degree C, and 0.4-0.8 log10 units over 12 weeks at -20 degrees C. The effect of incubation temperature upon viral inactivation rate was statistically significant (p less than 0.00001). As determined by pairing tests, survival was also significantly related to both viral serotype and water source at each of the three incubation temperatures (p less than or equal to 0.05). Efforts were made to determine whether the rate of viral inactivation observed at the different incubation temperatures was related to characteristics inherent to the water that was collected from the different locations. The characteristics examined included physical and chemical parameters, indigenous bacterial counts, and the amount of bacterial growth that the waters would support (measured as the maximum number of generations which seeded bacteria could undergo after being placed into either pasteurized or sterile-filtered water samples). Analysis of viral inactivation rate versus these characteristics revealed three apparent effectors of viral persistence.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545319 TI - Role of calcitriol in phosphate regulation by the chick embryo. AB - Chick embryos were injected on the 14th day of incubation with 100 ng calcitriol. The concentration of Ca in their serum rose significantly 4 hours after the injection and the concentration of Pi started to decrease 10 hours after. When embryos of the same age were injected with a solution containing CaCl2, the concentrations of both Ca and P rose significantly 2 hours after the injection and remained high until the end of the experiment. The fact that both treatments produced hypercalcemia but had opposite effects on the concentration of Pi does not agree with the idea that the hypophosphatemic response to calcitriol might be secondary to the hypercalcemia which precedes it. The injection of a solution of NaHCO3 to embryos of the same age failed to produce hypophosphatemia. The fact that calcium salts and bicarbonate, when injected separately, fail to induce hypophosphatemia does not contradict the possibility that the hypophosphatemic response to calcitriol might result from the simultaneous increase in flux of Ca and -HCO3 from the shell. Three days after the injection of calcitriol to 14-day old embryos, the total amount of Ca and P in the urine was significantly higher than in the controls. The concentration of Ca and P in kidney tissue was also significantly higher in the injected embryos. In addition, calcified precipitates were detected histochemically in the lumen of the kidney tubules from the treated embryos. These results are interpreted as demonstrating that an increase in the excretion of P in the urine is the main mechanism explaining calcitriol-induced hypophosphatemia in the chick embryo.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545322 TI - Production and characterization of neutralizing monoclonal antibodies against poliovirus type 1, 2, and 3. AB - Neutralizing monoclonal antibodies were produced against a reference vaccine or a reference wild strain of poliovirus type 1, 2, and 3. After 26 fusions, 55 monoclonal antibodies were obtained with serotype 1 as the immunizing antigen, 180 with serotype 2, and 115 with serotype 3. The neutralizing activity of these monoclonal antibodies was tested first with the two reference strains and then if reactive, against a panel of 10 well-characterized strains of each serotype, 5 vaccinelike (VL) and 5 nonvaccinelike (NVL). All monoclonal antibodies were type specific without reactivity with any of the heterologous strains. There was a wide range of reactivity within the strains of each serotype. Several monoclonal antibodies to serotype 1 reacted with all type 1 strains, while several neutralized strongly all VL strains and weakly one or more of the NVL strains. Most of the 180 monoclonal antibodies to serotype 2 neutralized to various degrees all strains of this serotype and about half reacted very strongly with all homologous strains whether VL or NVL. None could differentiate all VL and NVL homologous strains. Of the 115 monoclonal antibodies to serotype 3, several monoclonal antibodies neutralize to various levels all homologous strains and some can differentiate VL and NVL strains. PMID- 2545323 TI - Mortality rates among employees potentially exposed to chrysotile asbestos at two automotive parts factories. AB - A study of the mortality rates among 1657 employees at two Ontario automotive parts factories that manufactured friction materials containing chrysotile asbestos was initiated in response to the workers' concerns about the effects of asbestos on their health. A total of 1194 men and 258 women had had their first potential exposure at least 10 years before the end of the study period; 563 of the men and 138 of the women had had such an exposure at least 20 years before the end of the study period. A significantly increased rate of death from laryngeal cancer and an elevated rate of death from lung cancer were observed in a cohort analysis. One or two deaths might have been due to pleural mesothelioma. There was no increase in the rate of death from gastrointestinal cancer or from nonmalignant respiratory disease. Case-control analysis showed no association between the risk of laryngeal or lung cancer and the total duration of employment (a surrogate for the extent of ambient exposure to asbestos or other workplace toxic substances) or employment in departments where asbestos had been used. An association between risk of death and occupational exposure is uncertain. PMID- 2545324 TI - The development and extension of hepatohilar bile duct carcinoma. A three dimensional tumor mapping in the intrahepatic biliary tree visualized with the aid of a graphics computer system. AB - Computer-assisted three-dimensional (3D) reconstruction of the biliary tree including extrahepatic and intrahepatic parts was performed from surgical or autopsy materials from 12 patients with hepatohilar bile duct carcinoma in an effort to visualize three-dimensionally the distribution of carcinoma and dysplasia. In each case, material including a hepatic lobe was reduced to serial slices 1 mm thick using a ham slicer, then the tumors, ducts with carcinoma in situ, and those with dysplasia were submitted to reconstruction. In a 3D map of biliary tree reproduced on a display, a dysplastic zone was shown in most cases surrounding a focus of carcinoma, justifying the assumption of a dysplasia carcinoma sequence. The carcinoma itself proved to form multiple foci along bile ducts in as many as 42% of the patients, in some of whom the foci were independent without any intervening dysplasia. These results suggest that as extensive a surgical measure as possible should be taken in designing a strategy against this tumor. PMID- 2545325 TI - Small cell carcinoma of the bladder. Report of five cases with immunohistochemistry and review of the literature with evaluation of prognosis according to stage. AB - Five cases of small cell carcinoma (SCC) of the bladder are reported with pathologic and immunohistochemical findings and clinical follow-up. Sixteen additional cases reported in the literature are studied and staged according to depth of tumor infiltration of the bladder wall. In our series of five cases and those reviewed from the literature, survival appeared to be dependent on stage of the tumor, analogous to the more common transitional cell carcinoma (TCC). We report two of five patients with the longest recorded follow-up times in the literature who are alive with no evidence of disease at 4 and 6 years. Contrary to the belief that SCC of the bladder is an aggressive, rapidly disseminating tumor similar to SCC of the lung, our findings support the notion that this is a potentially curable neoplasm with a prognosis that appears to be dependent on stage and surgical resectability. PMID- 2545326 TI - Dermatofibrosarcoma protuberans with fibrosarcomatous areas. A clinicopathologic study of nine cases and a comparison with allied tumors. AB - Nine cases of dermatofibrosarcoma protuberans with fibrosarcomatous areas (DFSP with FS) were analyzed clinicopathologically, and the findings compared with allied tumors comprising 43 cases of ordinary DFSP, ten of superficial fibrosarcoma (SFS), and 11 of storiform variant of malignant fibrous histiocytoma (SV MFH). All but one case of DFSP with FS developed on the trunk of the body. Eight of the nine patients (89%) had one recurrence with a shorter prerecurrent interval than that of the ordinary DFSP, and a hipbone metastasis was identified in one patient. These findings reflect the unfavorable course, in contrast to the ordinary DFSP. Microscopically, the FS areas were presented either in the initial lesion (five cases) or in the recurrent one (four cases), areas showing cellularity, mitotic figures, and cytologic anaplasia that exceeded those in the DFSP areas. Dermatofibrosarcoma protuberans with FS and allied categories such as SFS and SV MFH shared various clinical and histologic features but there was a diversity in the prognosis relative to the grade of malignancy. The relationship between DFSP with FS and the allied tumors was given attention. PMID- 2545327 TI - Pleomorphic adenoma of the external auditory canal. An immunohistochemical and ultrastructural study. AB - Primary tumors arising from the glandular structures of the external auditory canal (EAC) are rare, with the majority arising from the ceruminous glands. The histochemical, immunohistochemical, and electron microscopic features of a pleomorphic adenoma of the EAC are presented; this is the seventh case reported. These studies confirm the key role of the myoepithelial cell in this tumor and include a correlation with pleomorphic adenomas in other sites. PMID- 2545329 TI - Alveolar rhabdomyosarcoma metastatic to the ovary. A report of two cases and a discussion of the differential diagnosis of small cell malignant tumors of the ovary. AB - Two young women with alveolar rhabdomyosarcoma metastatic to the ovary are reported. In each case, the ovarian involvement was detected within a few weeks of the discovery of a soft tissue mass by the patient. Symptoms attributable to the ovarian tumor dominated the initial clinical picture in one case. In the other case, the soft tissue mass was not appreciated by the treating physicians until after the diagnosis of rhabdomyosarcoma had been established by examination of the ovarian tumor. The ovarian tumor was unilateral in one case and bilateral in the other. One primary site was the right foot and the other primary site was the left forearm. These cases illustrate that metastatic alveolar rhabdomyosarcoma rarely enters into the differential diagnosis of a small cell malignant tumor of the ovary. Both patients experienced progression of their disease and died, despite aggressive chemotherapy, within 1 year of presentation. PMID- 2545328 TI - Glomus tumor of the trachea. Report of a case with ultrastructural observation. AB - Extracutaneous glomus tumors are unusual and their occurrence in the trachea has been recognized with extreme rarity. We present a case of surgically resected glomus tumor of the trachea in a 54-year-old woman who was initially misdiagnosed as having bronchial asthma because of expiratory wheezing. The tumor, located 7 cm below the vocal cord and 5 cm above the carina, was an intraluminally protruding polypoid mass that measured 1.5 X 1.2 cm and arose from the posterior wall of the trachea with a broad base. Microscopically, the tumor was predominantly of the solid type, but a small proportion of a glomangiomatous component was admixed. Ultrastructural study confirmed the presence of myofibrillar bundles with focal densities and fine pinocytotic vesicles along the plasma membrane. A structure similar to a nonneoplastic glomus was demonstrated in the submucosal layer of the posterior wall away from the tumor margin. This suggests that the tracheal glomus tumor occurs more often in the posterior wall where neuromuscular complex structures may develop more readily. PMID- 2545330 TI - Soft tissue fibrosarcoma. A clinicopathologic study of 132 cases. AB - Over the last 20 to 30 years, a recognition and general acceptance of the entity of malignant fibrous histiocytoma and monophasic synovial sarcoma has occurred. The introduction of these diagnostic categories among the generic group of spindle cell sarcomas now requires a reevaluation of the more traditional and historic entity of fibrosarcoma and the clinical ramification of this diagnosis. A review of 132 cases of fibrosarcoma using current diagnostic criteria was undertaken to clarify the behavior of fibrosarcoma as diagnosed in current patients. Special attempts were made to correlate histologic grade and Enneking stage in fibrosarcomas in an effort to correlate their predictive value in metastatic rate and survival. PMID- 2545331 TI - Germ cell function and hormonal status in patients with testicular cancer. AB - Sperm counts, serum gonadotropins, and androgen levels were investigated in 39 seminoma patients and 58 patients with a nonseminomatous germ cell tumor of the testis after unilateral orchiectomy. In 58% of the patients, the total sperm count was below the lower reference value (80 million). A multiregression analysis demonstrated a correlation between a decreased total sperm count and the following three explanatory variables: (1) an elevated serum alpha-fetoprotein (AFP), (2) a history of chryptorchidism, or (3) a seminomatous tumor. In 42% of the patients, the sperm concentration and the sperm motility met criteria considered sufficient for cryopreservation. Serum follicle-stimulating hormone (FSH) was elevated in 33% of the patients. Androgens (serum testosterone [T] or urine 17-oxy-steroids [17-OS]) were subnormal in 5% of the patients, whereas serum luteinizing hormone (LH) was elevated in 14% of the patients without human chorionic gonadotropin beta-subunit (beta-HCG) in serum. PMID- 2545332 TI - Differential effect of recombinant granulocyte macrophage colony-stimulating factor on human monocytes and alveolar macrophages. AB - The effect of granulocyte-macrophage colony-stimulating factor (GM-CSF), a pluripotent cytokine, on tumoricidal activity of alveolar macrophages and monocytes from nonsmoking normal volunteers was compared using [3H]thymidine labeled human tumor cells (SK-MEL-28, melanoma) as targets. A dose-response study (500-5000 units/ml) of recombinant GM-CSF indicated dramatic differences between cytotoxicity of alveolar macrophages and blood monocytes. Macrophages exhibited significant (P less than 0.01) tumoricidal activity at all GM-CSF doses tested. In contrast, monocytes showed no significant tumoricidal activity at 500 units/ml and significantly (P less than 0.01) less activity than alveolar macrophages at doses of 1000-5000 units/ml. Maximal activity in alveolar macrophages occurred 72 96 h after exposure to 1000-5000 units/ml GM-CSF. Tumoricidal activity may be related to the state of maturation, because monocytes matured in vitro for 7 days displayed enhanced tumoricidal activity after GM-CSF exposure. Tumor necrosis factor alpha and interleukin 1 beta were measured in supernatant fluids of 24-h GM-CSF-treated cells. No significant increase in either cytokine was detected after GM-CSF treatment of alveolar macrophages. Monocyte interleukin 1 beta secretion was not enhanced by GM-CSF; however, tumor necrosis factor alpha secretion was enhanced in some donors (three of five). Superoxide anion production of alveolar macrophages was not enhanced by GM-CSF. These data suggest that alveolar macrophage tumoricidal activity is induced by GM-CSF and is not dependent on oxidative metabolism or secreted forms of interleukin 1 beta or tumor necrosis factor alpha. PMID- 2545333 TI - Augmentation of lymphokine-activated killer cell cytotoxicity by monoclonal antibodies against human small cell lung carcinoma. AB - This study examined the lymphokine-activated killer (LAK) cell cytotoxicity on monoclonal antibody (MoAb)-bound tumor cells from the human small cell lung carcinoma cell lines H69 and H128. LAK cells were generated from normal peripheral blood mononuclear cells by incubation with interleukin 2 for 3 or more days. Cells from the LAK culture were cytotoxic to natural killer-sensitive (K562, 84% cytotoxicity) and natural killer-resistant (Daudi, 85%; H69 and H128, 69% and 97%, respectively) cell lines, and to freshly excised human lung (49%) and breast (57%) tumors. LAK cytotoxicity to H69 or H128 cells was significantly augmented by target cell preincubation with the small cell lung carcinoma reactive MoAbs 1096 (increases of up to 271%) or 5023 (up to 223%). SCLC 5023 or 1096 did not enhance LAK cytotoxicity to Daudi cells of lymphoblastoid origin. Pretreatment of LAK cells with an anti-Fc receptor antibody blocked MoAb augmentation by 1096 or 5023 (but not LAK cytotoxicity), suggesting that LAK-MoAb interaction may be mediated by Fc binding. LAK activity coincided with emergence of a large cell [interleukin 2-stimulated large mononuclear leukocyte (LML)] subset expressing the CD16 and NKH-1 surface determinants. Serial immunophenotyping of the LAK cell culture harvested at Days 3, 5, and 7 indicated that the level of LAK cytotoxicity, with or without MoAb augmentation, correlated with frequency of NKH-1-reactive LMLs. These observations support the hypothesis that LAK cytotoxicity is mediated by a NKH-1-reactive LML subpopulation. Antitumor cytotoxicity may be augmented by tumor-reactive MoAbs through Fc binding to this LML subset. PMID- 2545334 TI - Analysis of nerve growth factor receptor expression in human neuroblastoma and neuroepithelioma cell lines. AB - A series of 22 neuroepithelioma and neuroblastoma cell lines were screened for expression of nerve growth factor receptor (NGFR) by flow cytometry, Western blotting, and Northern blotting. All 5 neuroepithelioma cell lines expressed cell surface NGFR, with 30-69% of cells NGFR positive, but the 17 neuroblastoma cell lines tested had a smaller percentage of cell surface NGFR-positive cells (0-21%) and 10 lines were completely lacking cell surface NGFR. SY5Y, a variant line with a neuronal phenotype derived from neuroblastoma line SKNSH, expressed much more NGFR than SHEP, a variant line with an epithelial-like phenotype also derived from SKNSH. By Western blotting, the Mr approximately 69,000 NGFR band was detected for all four neuroepithelioma cell lines tested but was visible for only 8 of 15 neuroblastoma cell lines tested. The band was most intense for neuroepithelioma cell lines SKNMC and TC32. For these two lines, a Mr approximately 56,000 and a Mr approximately 60,000 band were also detected. By Northern blotting, all three neuroepithelioma cell lines tested were positive for the 3.8 kilobase NGFR mRNA, but only 8 of 15 neuroblastoma cell lines were positive. Neuroepithelioma cell line TC32 and neuroblastoma cell line GICAN had the strongest expression of NGFR mRNA. These results demonstrate that NGFR is a biological marker for neuroepithelioma and that NGFR expression is heterogeneous for neuroblastoma cell lines. This series of neural cell lines differing in NGFR expression will be useful for future studies of regulation of NGFR expression and neuronal differentiation. PMID- 2545336 TI - Differential expression of the gene encoding the novel pituitary polypeptide 7B2 in human lung cancer cells. AB - The protein designated 7B2 is a recently discovered pituitary polypeptide which is selectively expressed in cells containing secretory granules, such as neurons and endocrine cells. Northern blot analysis of 7B2 gene expression in small cell lung carcinoma (SCLC) cell lines revealed that 7B2 was expressed in all nine cell lines of the classic type tested, but in six of seven SCLC cell lines of the variant type, 7B2 expression could not be detected. In only one of four non-SCLC cell lines tested, 7B2 was expressed. Furthermore, in 16 primary human non-SCLCs, no or only very low expression of 7B2 was found. In the eight primary human SCLCs tested, expression of 7B2 appeared variable: three exhibited a high level of expression; three a low level; while in two cases, expression was very low or not detectable at all. Finally, the three carcinoid tumors tested expressed very high levels of 7B2 mRNA. These data indicate that the 7B2 gene is a useful marker not only to discriminate between classic and variant types of SCLC cell lines, but also in human lung cancer diagnosis. PMID- 2545335 TI - Dipyridamole enhancement of etoposide sensitivity. AB - Dipyridamole (DPM) enhanced the sensitivity of human ovarian carcinoma 2008 cells to etoposide (VP-16) producing a 5.5-fold reduction in 50% inhibitory concentration at a DPM concentration of 20 microM. This interaction was shown to be truly synergistic by isobologram and median effect analysis. DPM increased the steady-state VP-16 content of 2008 cells; a DPM concentration of 4 microM increased VP-16 content by 2-fold. DPM was 25 times less potent when cells were incubated in human plasma. In tissue culture medium 96% of the DPM was free, whereas in plasma only 15% was non-protein bound. DPM did not displace VP-16 from proteins under either condition. DPM did not increase the initial influx of VP-16 but did inhibit the initial efflux, reducing the efflux rate constant by 27%. DPM had no effect on the later stages of drug efflux, nor did it irreversibly bind VP 16 in the cell. The effect of DPM was evident within 1 min; once removed, the effect disappeared within 2 min. DPM is a potent nucleoside membrane transport inhibitor and can also inhibit cyclic AMP (cAMP) phosphodiesterase in platelets. Nitrobenzylthioinosine, another nucleoside transport inhibitor which competes for binding with DPM, did not enhance sensitivity to VP-16 or increase VP-16 cellular accumulation and did not block the effect of DPM. In 2008 cells, DPM did not increase cAMP; when cAMP was increased by incubation with dibutyryl cyclic 3':5' AMP, there was no synergy with VP-16. The results indicate that enhanced sensitivity to VP-16 was not due to an effect of DPM on the protein binding of VP 16 or on cellular cAMP and suggest that it is not directly related to inhibition of nucleoside transport. This effect appears to be a newly identified mechanism of action for this agent. PMID- 2545337 TI - Resistance mechanisms in three human small cell lung cancer cell lines established from one patient during clinical follow-up. AB - Mechanisms for resistance were studied in three classic type, human small cell lung cancer cell lines, GLC14, GLC16, and GLC19, that were established from one patient during clinical follow-up. Clinically the tumor changed from sensitive (GLC14) to completely resistant to (chemo)therapy (GLC19) during this period. The stain with JSB-1 antibody, detecting the Mr 170,000 multidrug resistance associated glycoprotein, was most pronounced in GLC16 and absent in GLC19. Intracellular Adriamycin (Adr) concentrations were decreased in GLC16 and GLC19 versus GLC14. Glutathione levels were 12.9, 15.5, and 16.6 micrograms/mg protein; total sulfhydryl groups were 36.5, 45.7, and 48.8 micrograms/mg protein; and glutathione S-transferase activity was 13, 29, and 43 nmol I-chloro-2,4 dinitrobenzene/min/mg protein for GLC14, GLC16, and GLC19, respectively. Incubation with DL-buthionine-S,R-sulfoximine increased Adr and cisplatin induced cytotoxicity, whereas X-ray induced cytotoxicity remained the same. Catalase activity increased from 0.88 to 1.73 to 3.83 mumol H2O2/min/mg protein in, respectively, GLC14, GLC16, and GLC19. Compared to GLC14 and GLC16, Adr induced a higher amount of DNA strand breaks in GLC19. In none of the three cell lines could Adr induced DNA strand breaks be repaired. X-ray induced a comparable amount of DNA strand breaks in all three cell lines but all cell lines were capable of repairing the X-ray induced DNA strand breaks within 90 min. It is concluded that a number of different mechanisms are operative and that some but not all of the observed changes in mechanisms for drug resistance in these lines correlate with the clinical data. PMID- 2545338 TI - Development of lymphoma in the thymus of AKR mice treated with the lymphomagenic virus SL 3-3. AB - A chronological study of the individual thymic lobes of young AKR mice after neonatal inoculation of the oncogenic AKR retrovirus SL 3-3 was performed. 100% of mice treated in this manner develop lymphoma between 60 and 100 days of age. A search for early lymphoma cells in individual thymi was carried out by inoculating the thymocytes subcutaneously in syngeneic and intrathymically in syngeneic and semisyngeneic recipients. Tumor progression was observed in animals between 48 and 60 days of age. These animals have: (a) normal weight lobes, in which no lymphoma cells could be detected, (b) thymus-dependent lymphoma cells, in one or both normal weight lobes; (c) thymus-independent lymphoma cells, found in lobes of normal weight as well as in thymi enlarged by lymphoma cells. Thymocyte characteristics of virus-treated animals of 21 to 63 days of age were compared with those of age-matched controls. Beginning at 28 days a concordant, progressive with time, increase of thymocyte surface staining for the viral envelope glycoprotein gp70 was seen in all lobes from virus-treated animals. Evaluation of cell surface markers by two-color fluorescence with antibodies to CD4 and CD8 showed that after 50 days of age, thymic lobes with and without lymphomas had nonspecific, but marked, alterations of the typical thymocyte surface marker pattern. No characteristic CD4, CD8 surface phenotype was found in primary lymphomas. Using probes for the T-cell receptor J beta 2 gene segments and the Akv ecotropic virus gp70 envelope genes, oligoclonality in J beta 2 rearrangements and clonality using the Akv env genes was demonstrated in thymi with the thymus-dependent phenotype. In lymphomas T-cell receptor beta gene probes showed either oligoclonality or clonality. Clonal virus integrations were found in these lymphomas. These experiments suggest the following series of events in virus-accelerated AKR lymphomagenesis. First, lymphoma cells arise which are initially thymus-dependent and can appear in one or simultaneously in both thymic lobes. These progress to become thymus-independent, fully autonomous, tumor cells. Thymocytes close to or at the time of the initial transformation event show a marked disorder of differentiation defined by the alterations in the CD4, CD8 surface phenotype distribution. PMID- 2545339 TI - Amplification of the integrated viral transforming genes of human papillomavirus 18 and its 5'-flanking cellular sequence located near the myc protooncogene in HeLa cells. AB - The human papillomavirus type 18 integrated in the HeLa cell genome is amplified on chromosome 8. E6, E7, and E1 open reading frames are amplified 5-fold, and the late viral DNA region, the viral long control region, and cellular flanking sequences are amplified 15-fold. The common 5'-flanking cellular DNA was localized by in situ hybridization of normal and HeLa cells only on chromosome 8 band q24. This flanking probe is included in the amplification unit of Colo320 cells, but in the case of HeLa the amplified region does not include the myc gene which is structurally conserved. Viral integration on chromosome 8 represents an independent event and not a rearrangement of viral DNA located on other chromosomes. The amplification of HPV-18 E6 and E7 open reading frames and the constitutive expression of the myc protooncogene may contribute to immortalization and/or proliferative capacity of HeLa cells. PMID- 2545340 TI - Acid pH in tumors and its potential for therapeutic exploitation. AB - Measurement of pH in tissue has shown that the microenvironment in tumors is generally more acidic than in normal tissues. Major mechanisms which lead to tumor acidity probably include the production of lactic acid and hydrolysis of ATP in hypoxic regions of tumors. Further reduction in pH may be achieved in some tumors by administration of glucose (+/- insulin) and by drugs such as hydralazine which modify the relative blood flow to tumors and normal tissues. Cells have evolved mechanisms for regulating their intracellular pH. The amiloride-sensitive Na+/H+ antiport and the DIDS-sensitive Na+-dependent HCO3-/Cl exchanger appear to be the major mechanisms for regulating pHi under conditions of acid loading, although additional mechanisms may contribute to acid extrusion. Mitogen-induced initiation of proliferation in some cells is preceded by cytoplasmic alkalinization, usually triggered by stimulation of Na+/H+ exchange; proliferation of other cells can be induced without prior alkalinization. Mutant cells which lack Na+/H+ exchange activity have reduced or absent ability to generate solid tumors; a plausible explanation is the failure of such mutant cells to withstand acidic conditions that are generated during tumor growth. Studies in tissue culture have demonstrated that the combination of hypoxia and acid pHe is toxic to mammalian cells, whereas short exposures to either factor alone are not very toxic. This interaction may contribute to cell death and necrosis in solid tumors. Acidic pH may influence the outcome of tumor therapy. There are rather small effects of pHe on the response of cells to ionizing radiation but acute exposure to acid pHe causes a marked increase in response to hyperthermia; this effect is decreased in cells that are adapted to low pHe. Acidity may have varying effects on the response of cells to conventional anticancer drugs. Ionophores such as nigericin or CCCP cause acid loading of cells in culture and are toxic only at low pHc; this toxicity is enhanced by agents such as amiloride or DIDS which impair mechanisms involved in regulation of pHi. It is suggested that acid conditions in tumors might allow the development of new and relatively specific types of therapy which are directed against mechanisms which regulate pHi under acid conditions. PMID- 2545341 TI - DNA topoisomerase I-mediated DNA cleavage and cytotoxicity of camptothecin analogues. AB - 20(S)-Camptothecin, the 20(S)-camptothecin sodium salt, and 12 analogues with substituents on the A ring differ widely in their effectiveness in the treatment of murine L1210 lymphoblastic leukemia in vivo. The drugs were screened in the following systems: System 1, the cleavage of DNA in the presence of purified topoisomerase I; System 2, drug-induced trapping of topoisomerase I in a covalent complex with DNA; and System 3, the induction of protein-associated DNA breaks in drug-treated L1210 leukemia cells. 9-Amino-20(S), 10-amino-20(RS), and 10,11 methylenedioxy-20(RS), drugs effective against murine L1210 leukemia in vivo, stabilize topoisomerase I-DNA cleavable complexes in a purified system and in cultured L1210 cells. Other analogues, inactive against L1210 leukemia in vivo, were totally ineffective in topoisomerase I-directed screens. The rest of the analogues were intermediate in terms of their antitumor and topoisomerase I directed activities. The study shows that the drug-induced accumulation of enzyme DNA cleavable complexes is directly proportional to drug cytotoxicity and antitumor activity. PMID- 2545343 TI - Isolation of two Chinese hamster ovary cell mutants hypersensitive to topoisomerase II inhibitors and cross-resistant to peroxides. AB - We have isolated two Chinese hamster ovary cell lines, designated ADR-4 and ADR 5, which exhibit hypersensitivity to intercalating agents and epipodophyllotoxins. These drugs are thought to exert their cytotoxicity via an interaction with the enzyme topoisomerase II. However, there is no apparent change in the level or catalytic activity of topoisomerase II in the mutant cells. Drug sensitivity does not appear to be due to increased drug transport because accumulation of radiolabeled actinomycin D is similar in mutant and wild type cells. Both mutant cell lines show enhanced resistance to hydrogen peroxide and to organic peroxides. ADR-4 cells show a degree of temperature sensitivity. ADR-5 cells show mild sensitivity to UV irradiation. Neither cell line shows significant sensitivity to mono- or bifunctional alkylating agents, ionizing radiation, or bleomycin. Cell fusion studies indicate that the phenotype of each mutant cell line is recessive and that the mutants represent two different genetic complementation groups. These studies also indicate that ADR-4 and ADR-5 Adriamycin-sensitive mutant, ADR-1. These results indicate that sensitivity to topoisomerase II inhibitors can result from abnormalities in several genes. These drug-sensitive mutants may be useful for studying the mechanisms of cell killing by topoisomerase II inhibitors, free radicals, and heat. PMID- 2545342 TI - Significant increase in the antibody to Gal alpha 1-3Gal structure in sera of patients with hepatocellular carcinoma after transcatheter arterial embolization. AB - Sera from the patients with chronic liver diseases and hepatocellular carcinoma (HCC) were tested for reactivity with neutral glycosphingolipids extracted from rabbit liver plasma membrane by enzyme-linked immunosorbent assay and thin-layer chromatography immunostaining. IgG class antibody to neutral glycosphingolipids was detected in 29.6% (8 of 27), 6.3% (1 of 16), 0% (0 of 8), 0% (0 of 25), and 6.9% (2 of 29) in the sera of patients with HCC, liver cirrhosis, autoimmune chronic active hepatitis, chronic hepatitis, and normal individuals, respectively. Using the serum positive for the antibody to neutral glycosphingolipids, the target antigen glycolipid was isolated. Negative ion fast atom bombardment mass spectrometry, exoglycosidases treatment, and permethylation analysis revealed that the main target antigen was IV3 alpha Gal-nLc4Cer. In enzyme-linked immunosorbent assay, IgG class antibody to IV3 alpha Gal-nLc4Cer was detected in 33.3% (9 of 27), 18.8% (3 of 16), 25% (2 of 8), 4% (1 of 25), and 6% (3 of 50) in the sera of patients with HCC, liver cirrhosis, autoimmune chronic active hepatitis, chronic hepatitis, and normal individuals, respectively. Of 9 HCC patients positive for the antibody, 6 had received transcatheter arterial embolization (TAE) therapy. Six of 10 patients who received TAE therapy had the antibody, whereas only 3 of 17 patients without TAE therapy had the antibody. This antibody may be a heterophile antibody, which recognizes Gal alpha 1-3Gal structure at the nonreducing terminal of the antigen. Since the occurrence of this antibody was closely related with TAE therapy, the necrosis of HCC induced by TAE therapy may stimulate the production of the antibody. PMID- 2545344 TI - Structure-dependent induction of aryl hydrocarbon hydroxylase in human breast cancer cell lines and characterization of the Ah receptor. AB - The structure-dependent induction of aryl hydrocarbon hydroxylase (AHH) and ethoxyresorufin O-deethylase by 2,3,7,8-tetrachlorodibenzo-p-dioxin, 2,3,7,8 tetrachlorodibenzofuran, and 1,2,4,7,8-pentachlorodibenzo-p-dioxin was determined in the MCF-7, T47-D, and MDA-MB-231 human breast cancer cell lines. Both the MCF 7 and T47-D cells were responsive to the induction effects of the halogenated aryl hydrocarbons and the structure-induction relationships were comparable to the reported structure-activity (induction, receptor binding, and toxicity) relationships observed in rodents and rodent cells in culture. The induction of ethoxyresorufin O-deethylase in the T47-D cells was the most sensitive aryl hydrocarbon (Ah) receptor-mediated response in both cell lines and this enzyme activity was more inducible than aryl hydrocarbon hydroxylase. In contrast, the three congeners were inactive as monooxygenase enzyme inducers in the MDA-MB-231 cells. Despite the differential Ah responsiveness of the cell lines, incubation of the cells with [3H]-2,3,7,8-tetrachlorodibenzo-p-dioxin followed by extraction of the nuclei with high salt and velocity sedimentation analysis of the extracts showed that specifically bound nuclear Ah receptor complexes were present in the three cell lines. The sedimentation coefficients (and levels) for the nuclear receptors were 6.6 S (32.1 fmol/mg protein/mg DNA), 6.9 S (61.6 fmol/mg protein/mg DNA), and 7.4 S (38.2 fmol/mg protein/mg DNA) in the T47-D, MCF-7, and MDA-MB-231 cell lines, respectively. Cytosolic receptor was also detected in the MCF-7 and MDA-MB-231 cells. Thus, despite the differences in Ah responsiveness of the T47-D and MDA-MB-231 cells, comparable levels of nuclear receptor were detected in both cell lines. Furthermore, the elution profiles of the nuclear receptors from DNA-Sepharose columns by using a salt gradient were similar and this suggested that defects in the DNA-binding activity of MDA-MB-231 nuclear receptor complexes were not major factors associated with their failure to respond to 2,3,7,8-tetrachlorodibenzo-p-dioxin and related compounds. PMID- 2545345 TI - ABH blood group antigen expression, synthesis, and degradation in human colonic adenocarcinoma cell lines. AB - The synthesis of blood group ABH antigens is under genetic control, where the primary gene products are glycosyltransferases. Several studies have demonstrated cancer-associated alterations in ABH antigen expression in human colon cancer tissues. However, the mechanism(s) responsible for these alterations has not been elucidated. Therefore, experiments were conducted using nine established colon cancer cell lines (four type O, three type A, and two type B) to examine ABH antigen expression by immunocytochemistry and correlate this with activities of ABH biosynthetic (glycosyltransferase) and degradative (glycosidase) enzymes. The products of the glycosyltransferase enzymes were characterized by high performance liquid chromatography and paper chromatography, and substrate affinities (apparent Km values) of the cancer cell-derived glycosyltransferases were analyzed. The present data demonstrate: (a) all cell lines except H-498 (blood type A) expressed the appropriate ABH glycosyltransferase as well as all three glycosidases; (b) product characterization and substrate dependence experiments suggested that the cancer cell-derived ABH glycosyltransferase enzymes had properties that were similar to those of the ABH enzymes in human serum; (c) H-498 cells exhibited A antigen deletion with accumulation of H precursor substance, most likely due to insufficient A transferase activity; (d) SW1417 cells (blood type B) demonstrated B antigen deletion without precursor accumulation, despite adequate levels of B transferase and low alpha galactosidase activity; and (e) weak incompatible A antigen expression occurred in LoVo (type B) and SW1116 (type O) cells, and weak incompatible B antigen expression occurred in H-498 (type A) and SW1116 cells. However, since these cells lacked incompatible A or B transferase activity, these incompatible antigens are probably not the true A or B antigens. Thus, the colon cancer cell lines used in this study exhibit all of the ABH alterations previously described in colon cancer tissues and appear to be useful experimental models for studying the molecular events involved in cancer-associated ABH expression. PMID- 2545346 TI - Intracellular distribution and pharmacokinetics of daunorubicin in anthracycline sensitive and -resistant HL-60 cells. AB - Anthracycline-sensitive (HL-60) and -resistant (HL-60/AR) cells, which do not overexpress the P-glycoprotein, each transport and distribute daunorubicin (DNR) into distinct intracellular locations, as visualized by digitized video fluorescence microscopy. At pH 7.4, the fluorescence of DNR in HL-60 cells appears distributed diffusely in both the nucleus and cytoplasm. In contrast, HL 60/AR cells show much less fluorescence in the nucleus and cytoplasm; most of the fluorescence localizes first to the Golgi apparatus and is then gradually shifted to the lysosomes and/or mitochondria. In pharmacokinetic studies, HL-60/AR cells exposed to different extracellular concentrations of [14C]DNR consistently accumulated less radioactive drug than the parent HL-60 cells. Incubation of HL 60/AR cells with sodium azide and deoxyglucose blocked the efflux of [14C]DNR and also prevented the shift of DNR fluorescence from the Golgi apparatus to the lysosomes/mitochondria. The efflux and the intracellular shift of DNR could also be inhibited by lowering the temperature to 18 degrees C, which stops endosomal membrane fusion. When DNR was allowed to accumulate in HL-60 or HL-60/AR cells at pH 5 there was an increase in the proportion of drug fluorescence in the membranes of both HL-60 and HL-60/AR cells; a decrease in the amount of drug retained by HL-60, but not by HL-60/AR cells; and a decrease in the cytostatic effects of DNR on both HL-60 and HL-60/AR cells. These data suggest that DNR resistance is associated with a failure of DNR to pass through membranes and to bind to cytoplasmic and nuclear structures. Instead, most of the drug is taken up by the Golgi apparatus from which it is then shifted to the lysosomes or to mitochondria, or out of the cell. PMID- 2545347 TI - Expression of type IV collagenase and procollagen genes and its correlation with the tumorigenic, invasive, and metastatic abilities of oncogene-transformed human bronchial epithelial cells. AB - In a series of immortalized human bronchial epithelial cell lines continuing various oncogenes, the transcriptional levels of the type IV collagenase and the type IV procollagen genes were compared with the properties of invasiveness in vitro and tumorigenicity and metastatic ability in athymic nude mice. v-Ha-ras greatly enhanced invasion and metastasis, whereas v-Ki-ras, c-myc, and c-raf had lesser effects on these malignant phenotypes. In addition, cell lines derived from tumors obtained by injecting the original immortalized human bronchial epithelial cell lines into nude mice exhibited enhanced invasive and metastatic abilities and increased level of type IV collagenase mRNA when compared with the original immortalized human bronchial epithelial cell lines. Invasiveness and metastatic capacity correlated positively with expression of the type IV collagenase gene and negatively with the expression of the type IV procollagen gene, suggesting that these phenotypes are associated both with decreased production and increased dissolution of extracellular matrix. PMID- 2545348 TI - Biochemical epidemiology of colon cancer: effect of types of dietary fiber on fecal mutagens, acid, and neutral sterols in healthy subjects. AB - Several epidemiological studies suggest an inverse relationship between fiber intake and colon cancer risk. Animal model studies indicate that this inhibitory effect depends on the source of dietary fiber. Because of the potential significance of certain colonic mutagens and secondary bile acids in the pathogenesis of colon cancer, the effect of types of supplemental fiber on fecal mutagens and bile acids was studied in human volunteers. Seventy-two healthy individuals consuming high-fat/moderately low-fiber diets were screened for fecal mutagenic activity using the Ames Salmonella typhimurium/microsomal assay system. Twenty-one of them were found to excrete high levels of mutagens, and 19 of them were recruited for the diet intervention study. All participants provided two 24 h stool specimens and a 4-day food record while consuming their normal (control) diet. They were then asked to consume the control diet plus 10 g of dietary fiber from wheat bran, oat fiber, or cellulose for 5 wk. After each fiber period, they were asked to consume their control diet. At the end of each fiber and control diet period, each subject provided two 24-h stool specimens. Stool samples were analyzed for bile acids and mutagens using the Ames strains TA98 and TA100 with or without S9 (microsomal) activation. The concentrations of fecal secondary bile acids (deoxycholic acid, lithocholic acid, and 12-ketolithocholic acid) and of fecal mutagenic activity in TA98 and TA100 with and without S9 activation were significantly lower during the wheat bran and cellulose supplementation periods. Oat fiber supplementation had no such effect on these fecal constituents. Thus, the increased fiber intake in the form of wheat bran or cellulose may reduce the production and/or excretion of mutagens in the stools and decrease the concentration of fecal secondary bile acids in humans. PMID- 2545349 TI - High-dose cyclophosphamide or melphalan with escalating doses of mitoxantrone and autologous bone marrow transplantation for refractory solid tumors. AB - As the dose-limiting toxicity of mitoxantrone is hematological, the drug is suitable for dose escalation and use in intensive chemotherapy followed by autologous bone marrow rescue. Adult patients with therapy-resistant solid tumors received a regimen of high-dose cyclophosphamide (7 g/m2) and escalating doses of mitoxantrone in dose steps of 30, 45, 60, and 75 mg/m2. Both drugs were given i.v. on 3 consecutive days. Despite the addition of mesnum (3.5 to 7 g/m2), hemorrhagic cystitis occurred on the second day in four of eight patients, irrespective of the mesnum or mitoxantrone dose. Therefore, the cyclophosphamide in the combination regimen was replaced by high-dose melphalan (180 mg/m2). Mucositis was dose limiting at 75 mg/m2 of mitoxantrone. Responses were seen in eight of ten evaluable patients with four complete responses. Three responders received, after the autologous bone marrow transplantation program, radiotherapy or surgery on pretreatment bulky tumor localizations. Five patients still have disease-free survival after 9 to 36 mo. Pharmacokinetic studies of mitoxantrone were performed by high-performance liquid chromatography with UV detection. The plasma disappearance of mitoxantrone fitted into a three-compartment model with a mean t1/2 alpha of 10 min, a mean t1/2 beta of 96 min, and a slow elimination phase of 172 h. The mean distribution volume was 4294 +/- 3836 liters. We conclude that the high-dose cyclophosphamide-mitoxantrone regimen led to unexpected bladder toxicity, but the combination of melphalan (180 mg/m2) and mitoxantrone (60 mg/m2) can probably be given without major extramedullary toxicity. However, more patients should be evaluated at this dose before definite conclusions can be drawn about toxicity. PMID- 2545350 TI - Changes in the number of beta-adrenergic receptors of lymphocytes in patients with varying degrees of congestive heart failure. AB - This study was designed to determine changes in the number of beta-adrenergic receptors of circulating lymphocytes in patients with varying degrees of congestive heart failure. Thirty-seven patients with congestive heart failure were classified into four groups according to the functional classifications of the New York Heart Association (class I, 12 patients; class II, 12 patients; class III, eight patients; class IV, five patients). The control group consisted of 24 healthy volunteers. Binding assays of beta-adrenergic receptors in circulating lymphocytes and plasma norepinephrine concentrations were determined in each group. Compared with the control group, the number of beta-adrenergic receptors in patients of the class-III and class-IV groups was reduced by 37% and 36%, respectively, and plasma norepinephrine concentrations of class-III and class-IV patients were increased by 3.5 times and 5.9 times, respectively. No significant changes in the numbers of beta-adrenergic receptors or in plasma norepinephrine concentrations were observed in patients of the class-I and class II groups. These findings show a decrease in the number of beta-adrenergic receptors in patients with high plasma norepinephrine concentrations. It is suggested that the chronically high concentration of norepinephrine, resulting from activation of the sympathetic nervous system associated with severe congestive heart failure, is linked with the down regulation of beta-adrenergic receptors. The action of sympathomimetic amines may therefore be altered in patients with congestive heart failure. PMID- 2545351 TI - Stimulation of Na+,K+-ATPase activity in certain membranes of the rat central nervous system (CNS) by acute administration of desipramine (DMI). AB - 1. The activities of ATPase in rat CNS were studied 3 hr after administration of the noradrenaline uptake inhibitor, desipramine (DMI: 10 mg.kg-1, i.p.). Na+K+ ATPase activity significantly increased after DMI in the whole particulate from hypothalamus and mesencephalus but no changes in frontal cortex or in pons medulla oblongata areas were found. This increase was prevented when the animals were pretreated with the noradrenergic neurotoxic N-(2-chloroethyl)-N-ethyl-2 bromobenzylamine (DSP-4). 2. Purified membrane fractions from hypothalamus were obtained by differential and sucrose gradient centrifugation (0.8-1.2 M sucrose). It was observed that after DMI, Na+,K+-ATPase activity increased only in the membranous fraction lying at 0.9 M sucrose. 3. Mg2+- or Ca2+-ATPase activities were not modified by DMI treatment. 4. Citalopram, a specific serotonergic uptake inhibitor, did not affect ATPase activities. 5. The results obtained could indicate that DMI acute administration selectively stimulates Na+,K+-ATPase activity of certain membranes of the CNS after an increase in the concentration of the noradrenergic neurotransmitter in the synaptic gap. PMID- 2545352 TI - [Therapeutic and preventive care of trophoblastic disease in Czechoslovakia]. AB - The authors list units of trophoblastic disease and its classification used and elaborated in the Centre for trophoblastic disease. The authors submit detailed information on the organization of diagnostic and therapeutic and preventive care of patients with trophoblastic disease in the CSSR. The authors draw attention to the establishment of a Centre for trophoblastic diseases (CTN) with the statute of a reference department. Reasons for its legalization are given. Its function and structure are described. PMID- 2545353 TI - The effect of low- and high-density lipoprotein cholesterol on steroid hormone production and ACTH-induced differentiation of rat adrenocortical cells in primary culture. AB - We studied the effects of lipoprotein-derived cholesterol on the ACTH-induced differentiation of cultured fetal rat adrenocortical cells. For this purpose human plasma high-density lipoprotein3 (HDL3) or low-density lipoprotein (LDL) was added to culture media devoid of cholesterol, and thereafter the morphological changes in cells were monitored and the amounts of steroids synthesized were measured. It could be demonstrated that, ultrastructurally, upon ACTH-stimulation the adrenocortical cells differentiated into fasciculata-like cells even in the absence of lipoproteins in the culture medium. The addition of either HDL3 or LDL caused an increase in the number and size of cytoplasmic lipid droplets suggesting uptake and deposition of lipoprotein-derived cholesterol into the differentiating cells. The amount of steroids secreted from cells differentiating in media devoid of cholesterol was only half that observed in cells differentiating in serum-supplemented medium. Addition of either HDL3 or LDL increased the ACTH-stimulated steroid synthesis to the levels observed in serum-supplemented medium. This study demonstrates that both HDL3 and LDL are able to provide cholesterol for steroid synthesis accompanying the ACTH-induced differentiation of fetal rat adrenocortical cells. PMID- 2545354 TI - Immature rat epididymal epithelial cells grown in static primary monolayer culture on permeable supports. II. Histochemistry and ultrastructure. AB - N-acetylglucosaminidase (NAG), acid phosphatase (ACP) and alkaline phosphatase (AKP) were localised histochemically in fixed cells from the 37-day-old rat epididymis grown in static monolayer culture for 2-8 days. ACP and NAG were cytosolic enzymes found in perinuclear positions, whereas staining of AKP ws consistent with a membranous position. These enzymes were also examined in frozen tissue sections of the epididymis, from rats of the equivalent age, where NAG had intense activity in both supra- and infra-nuclear cytoplasm and ACP was more active apically. For the first time AKP was localised along basolateral membranes of the epithelium and in the lumen of the mid-caput region. The monolayer in culture was of principal cells only and they maintained their polarity and ultrastructural characteristics, but the height of the cells was reduced compared to that obtained in situ. PMID- 2545356 TI - Secretion of Mr 46,000 protein from human hepatoma cells treated with tumor promotors is regulated by c-myc gene expression. AB - Previously an Mr 46,000 protein (named p46) was shown to be induced in the culture medium of human hepatoblastoma cells, Huh-6 Cl-5, treated with 12-O tetradecanoyl phorbol-13-acetate (TPA). For further characterization of p46, Huh 7 Cl-4, another line of well differentiated liver cells, was incubated in the presence and absence of TPA, and proteins were labeled with [35S]methionine, and the proteins secreted into the medium were analyzed by one- and two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis. A protein of Mr 46,000 was observed in the culture medium of Huh-7 Cl-4 cells treated by TPA or plated at low density, but only slightly in the culture medium of Huh-7 Cl-4 cells plated at high density. As these results suggested that the expression of p46 was regulated by a factor that was activated in the growth state and by TPA treatment, the effect of introduction of the competence gene c-myc into Huh-7 Cl 4 cells was examined. All 37 independent transfected colonies obtained showed enhanced expression of p46. As a control, the c-Ha-ras gene was introduced into Huh-7 Cl-4 cells and shown not to enhance expression of p46. These results strongly suggested that the expression of p46 was regulated by the competence gene c-myc. PMID- 2545355 TI - Immunocytochemical and ultrastructural study of the frog (Rana pipiens) pars distalis with special reference to folliculo-stellate cell function during in vitro superfusion. AB - The colloidal gold immunocytochemical technique was used to determine the ultrastructural features of the glandular cells in the pituitaries of male frogs, Rana pipiens, both in vivo and after superfusion in vitro. Specific reactions to antisera against bullfrog gonadotropins, human prolactin, and synthetic 1-39 corticotropin allowed identification of the 3 corresponding types of glandular cells. No immunoreaction was obtained with antisera against human or ovine-growth hormone, human beta-thyrotropin hormone, and bovine S-100 protein. General morphological features of these immunocytochemically identified glandular cells were similar to those of equivalent cells previously described in other amphibian species. Non-glandular folliculo-stellate cells were distinctive. In freshly removed pituitaries, these folliculo-stellate cells contained lysosome-like structures, but did not show phagocytic vacuoles in the cytoplasm; they contained many mitochondria, and the Golgi complex and endoplasmic reticulum were relatively undeveloped. After 4 or 18 h of superfusion, some immunoreactive gonadotropic, prolactin, and corticotropic cells showed degeneration and destruction. In the same gland, folliculo-stellate cells retained a viable appearance, but showed phagocytic vacuoles containing secretory granule-like structures which were immunoreactive to gonadotropic, prolactin, and corticotropic antibodies. Some folliculo-stellate cells showed phagocytic vacuoles containing complete glandular cells. These results suggest that superfusion causes a destruction of some of the glandular cells, and that folliculo-stellate cells act as phagocytes when cellular debris or moribund cells are present in the intercellular space in the pituitary parenchyma. PMID- 2545357 TI - Structure and expression of a split chloroplast gene from mustard (Sinapis alba): ribosomal protein gene rps16 reveals unusual transcriptional features and complex RNA maturation. AB - The mustard chloroplast gene rps16 is split by an 887 bp group II (or III) intron. Three RNA 5' ends upstream of the rps16 coding region define both the transcription start site and two RNA processing sites. The DNA region preceding the start site contains a procaryotic-type "-10" promoter element, but not a typical "-35" element. One single RNA 3' end has been detected downstream from the rps16 coding region, but it is not in close proximity to any inverted repeat that might serve as a termination signal. Northern analysis has revealed several rps16 transcripts ranging in size from 1.6 kb to 0.5 kb. During seedling development, transcript levels show an initial increase and then remain constant without much difference between seedlings grown under light or in the dark. PMID- 2545358 TI - Increased diuron resistance in the joint expression of mutations located at the DIU2, DIU3 and DIU4 loci of Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, diuron blocks the respiratory pathway at the level of the bc1 complex. Two mitochondrially inherited loci, DIU1 and DIU2, located in the cytochrome b gene, and two nuclearly inherited loci, DIU3 and DIU4, have previously been identified. The present work genetically characterizes two double mutants. One mutant, Diu-217, carries two nuclearly inherited mutations, diu3 217a and diu-217b; the second mutant, Diu-783, carries the previously described nuclear mutation diu3-783 and a mitochondrial mutation diu2-783. Each mutation, independent of its location, exhibits a weak diuron resistance. The joint expression of two or three mutations leads to a cumulative or a cooperative enhanced diuron-resistant phenotype. PMID- 2545359 TI - [Malignant fibrous histiocytoma of the left atrium of the heart]. AB - Malignant fibrous histiocytoma was found in the left heart atrium as well as in probable secondaries in stomach, intestine, calvaria and diaphysis of femur. Tumour cells possessed electron microscopical features and markers of fibroblasts and histiocytes (immunopositivity with lysozyme,i A1 AT and A1 ACT). PMID- 2545360 TI - [Transcranial and spinal stimulation of motor pathways using conventional stimulator]. AB - In a group of 31 subjects with a normal locomotor system in the area of the upper extremities transcutaneous cortical and spinal stimulation was administered using a conventional stimulator with an electromyographic tracing on the contralateral tenar and a common investigation of the motor conduction of the median nerve. The mean latency on stimulation on the calva was 20.0 +/- SD 0.9 ms, the mean amplitude 3.8 +/- SD 1.6 mV. The conduction time between cortex and the cervical spinal area was on average 4.8 +/- SD 0.5 ms, between cortex and wrist 16.26 +/- SD 0.8 ms, the mean latency on spinal stimulation 15.2 +/- SD 0.9 ms. The authors found a statistically significant relationship between the latencies from the calva, and spinal stimulation, with the height of the subject and with the length of the upper extremity, while there was no relationship between the conduction time in the central portion of the motor pathway and these anthropometric parameters. The authors evaluate the risk and tolerance of transcranial stimulation in man. PMID- 2545361 TI - [Adherence of mononuclear leukocytes in multiple sclerosis: the effect of the calcium ionophore A 23187 and leukotrienes B4 and C4]. AB - The author assessed the effect of leucotrienes B4 and C4 and the calcium ionophore A23187 on the adherent reactivity of mononuclear leucocytes in patients with multiple cerebrospinal sclerosis and in controls. The calcium ionophore A23187 in concentrations of 200 nmol/1 - 5 mumol/1 induced inhibition of leucocyte adherence, depending on the concentration used. This reactivity was, as compared with controls, significantly lower in patients with multiple sclerosis who did not take cortisoid preparations and in patients taking prednisone in doses up to 5 mg/day. In patients treated with higher prednisone doses (20-90 mg/day) the values of the non-adherence index reached values insignificantly higher than in controls. Leukotrienes B4 and C4 (1 nmol/1 - 100 nmol/1 stimulated leucocyte adherence in patients treated with higher corticoid doses, in the remaining patients and in controls they did not influence the leukocyte adherence. The results of the investigation provide evidence of abnormalities of the membrane properties of the mononuclear leucocytes in patients with multiple sclerosis and draw attention to the association of these changes with metabolic chains of arachidonic acid and with calcium ion homeostasis. PMID- 2545362 TI - Lithium chloride SCN injection alters the circadian rhythm of food intake. AB - Effect of lithium injections through chronically implanted cannulae into the bilateral suprachiasmatic nuclei (SCN) on the circadian rhythm of food intake was investigated in the rat. It was observed that the circadian rhythm was disrupted by injections of lithium at the beginning of the light as well as the dark phase of the LD cycle. In either case the percentage of the food consumed during the 12 hr light period increased while that during the dark period decreased without any significant change in the total daily intake. Disruptions in the circadian rhythm of food intake failed to show any dose-response relation. Injections of saline into the SCN or lithium into the nearby SCN area did not produce a disruption of the circadian rhythm of food intake. PMID- 2545363 TI - Involvement of cysteine, serotonin and their analogues in peroxidase-oxidase reactions. AB - Myeloperoxidase-oxidase reactions with close to physiological concentrations of thiols and phenols were studied. Cysteine was shown to be a myeloperoxidase oxidase substrate when catalytic amounts of serotonin were added as cosubstrate. Penicillamine could be substituted for cysteine and acetaminophen could be substituted for serotonin. The properties of these peroxidase-oxidase reactions, e.g. the dependence on substrate and myeloperoxidase concentration, reduced oxygen species, metal ions and pH, were studied. Also, eosinophil, lacto- and horseradish peroxidase could catalyse these reactions. PMID- 2545364 TI - Effects of Ca2+ and V5+ on glucose-6-phosphatase activity in rat liver microsomes: the Ca2+ effect is reversed by regucalcin. AB - The effect of regucalcin, a calcium-binding protein isolated from rat liver cytosol, on glucose-6-phosphatase in the microsomes of rat liver was investigated. Addition of Ca2+ up to 2.5 microM to the enzyme reaction mixture caused a significant increase of glucose-6-phosphatase activity in hepatic microsomes, while Ni2+, Zn2+, Cd2+, Cu2+, Mn2+ and Co2+ (20 microM) did not have an appreciable effect. Vanadate (V5+) markedly inhibited the enzyme activity; a significant inhibitory effect was seen at 10 microM V5+. The Ca2+-induced increase of glucose-6-phosphatase activity was reversed by the presence of regucalcin; the effect was complete at 1.0 microM of the protein. Regucalcium had no effect on the basal activity of the enzyme. Meanwhile, the inhibitory effect of V5+ (10-100 microM) on glucose-6-phosphatase was not appreciably blocked by the presence of regucalcin (up to 2.0 microM). The present data suggest that hepatic microsomal glucose-6-phosphatase is uniquely regulated by Ca2+ and V5+, of various metals, and that the Ca2+ effect is reversed by regucalcin. The present study supports the view that regucalcin plays an important role as a regulatory protein in liver cell function related to Ca2+. PMID- 2545365 TI - Epstein-Barr virus: the hematologic and oncologic consequences of virus-host interaction. AB - Varicella-zoster virus (VZV) and Epstein-Barr virus (EBV) are two of the human herpesviruses. The others include herpes simplex virus (HSV) type 1, HSV type 2, and cytomegalovirus (CMV). In a series of two articles, we review the clinical diseases caused by VZV and EBV infections; we pay particular attention to the manifestations of these two viral infections in immunosuppressed and immunocompromised patients. In addition to the clinical reviews, each of the two articles begins with a brief discussion of the molecular aspects of VZV and EBV, respectively; this introduction describes features of the genome and immunogenic viral proteins which have clinical relevance. A model for pathogenesis is included. The first review concerns VZV infections. Recent data about the DNA sequence of the entire VZV genome are included, as well as a review of the VZV glycoproteins. Primary VZV infection (chickenpox) and VZV reactivation (zoster) are described in detail in both healthy individuals and people with cancer. The decade-long VZV vaccine trials in children with leukemia receive special emphasis because they have engendered considerable interest and debate. The second review (published here) covers EBV infections. This virus has been implicated in the causation of a wide variety of human hematological and oncological disorders, besides classical infectious mononucleosis. In particular, Burkitt's lymphoma, nasopharyngeal carcinoma, and lymphoproliferative disorders are strongly associated with EBV infection of the transformed cells. In addition, immunologically mediated cytopenias occasionally follow EBV infection. Finally, treatment regimens with antiviral chemotherapy and other agents are discussed for both VZV and EBV infections. PMID- 2545366 TI - [A study in the Ivory Coast (1985-1987) of the efficacy and tolerance of ivermectin (Mectizan) in human onchocerciasis. I. A comparative double-blind study of 220 patients with onchocerciasis treated with a single oral dose of 100, 150 or 200 mcg/kg]. AB - Two hundred and twenty adult males living in endemic onchocerciasis areas in Ivory Coast, with a mean of 59 to 64 mf/mg of skin microfilariae, having or not ocular lesions, were divided into 4 homogeneous groups and treated by placebo or by a single oral dose of 100, 150 or 200 mcg/kg of ivermectin. Parasitological, clinical, ophthalmological, biological data were gathered before treatment (J less than 1) and at day 4 and 3, 6, 12 months post treatment. The doses of 150 to 200 mcg/kg gave the best results with a reduction of microfilariae of 75 to 79% at day 4 and of 97 to 99% at 3 months. A slight increase appears at six months reaching to 10% of the initial level, at 12 months. These posologies reduce also the number of ocular microfilariae 12 months after treatment. The percentage of patients presenting microfilariae in the cornea varies from 4 to 14% according to the given dose whereas it was initially between 26 and 33%, and in the anterior chamber from 22 to 16% instead of 62 to 67%. In patients who were still positive after treatment the detected number of ocular microfilariae was very small. Side effects starting 12 to 24 hours after treatment are similar to those appearing during the normal evolution of onchocerciasis. They were observed in 36% of subjects receiving a placebo and 56 to 65% of treated subjects. Statistically they are neither correlated with the intensity of parasitism nor to the given posology and disappear spontaneously few days later or after administration of aspirin and/or antihistaminic. Ivermectin given at a single oral dosage of 150 to 200 mcg/kg is a powerful microfilaricidal drug with a rapid and prolonged action and without major side effects. PMID- 2545367 TI - [A study in the Ivory Coast (1985-1987) of the efficacy and tolerance of ivermectin (Mectizan) in human onchocerciasis. II. Evaluation in the light of mass campaigns on the effect of yearly or half-yearly administration of single oral doses of 100, 150 or 200 mcg/kg]. AB - One hundred and twenty patients out of 220 in a previous study were retreated after 6 months or one year with doses of 100, 150 or 200 mcg/kg of ivermectin. The tolerance was excellent due probably to the small number of skin microfilariae obtained with the first treatment. The annual treatment with 150 to 200 mcg/kg, better than with 100 mcg/kg, keeps for the year following the second treatment the microfilariae number between 6 and 11% of the initial level. The half-yearly administration keeps it between 1 and 7% and especially 94 to 100% of these retreated patients have a level of microfilariae less than 5 mf/mg. The results obtained with the half-yearly treatment show a considerable reduction of the number of microfilariae in the anterior chamber of the eye and the percentage of positive patients. Ivermectin is a very efficient microfilaricidal drug for the treatment of onchocerciasis and the prevention of ocular complications. Its rational use in mass campaigns should reduce, if not interrupt, the transmission of this parasitic disease. PMID- 2545368 TI - [A study in the Ivory Coast (1985-1987) of the efficacy and tolerance of ivermectin (Mectizan) in human onchocerciasis. III. The tolerance and efficacy of a single oral dose of 150 mcg/kg in children]. AB - One hundred and three male and female children of 6 to 14 years old with onchocerciasis, having or not ocular involvement and a mean skin microfilariae level of 36.1 mf/mg, received, in October 1986, a single oral dose of 150 mcg/kg ivermectin and controlled at day 4, 3 months, 6 months and 12 months post treatment. After the last control they were retreated with the same dose. The skin microfilariae count fell down to 94% of the initial level at day 4 and to 99% at 3 months. At 6 months the microfilariae count was 2% of the initial level and 5% at 12 months. The percentage of patients having microfilariae in the anterior chamber of the eye which was 33% before treatment fell down to 6% at 12 months. The percentage of patients having microfilariae in the cornea was 39% before treatment and 18% at 12 months. In the cornea and anterior chamber there was a very reduced number of microfilariae still present. 65% of the children had lesions of keratitis before treatment and 34% two months later. Adverse effects (fever, headache, pruritus, oedemas, myalgias, arthralgias) occurred in 64% of children after the first treatment and 50% after the second. They were of weak or moderate intensity and receded rapidly after administration of aspirin and/or anti-histaminic. The administration of ivermectin is an efficient and well tolerated drug in children above 5 years old. PMID- 2545370 TI - [Screening for urinary schistosoma by strips reactive to hematuria. Evaluation in zones of intermediate and weak endemicity in the Ivory Coast]. AB - Sticks Hemastix, reactive to haematuria have been tested into a focus ivory coast located in a humid savanna area. Into this focus the prevalence rate of urinary schistosomiasis (Schistosoma haematobium) was 27% taken as a whole and 40% for 5 to 19 years old persons. This study has shown that reactive sticks have a good sensitivity and an acceptable specificity compared with filtration. More, a good correlation between intensity of eggs count and the haematuria is observed. PMID- 2545369 TI - [Pruritus after taking chloroquine and filariasis]. AB - A study of relations between pruritus after doses of chloroquine and on the one hand the infection by Mansonella perstans (filariasis frequent in some areas but pathogenic) and on the other hand the infection by Onchocerca volvulus (filariasis frequent in some other areas but not very pathogenic) was carried out in two villages in Ivory Coast. It is not possible to show an obvious relation between the presence of pruritus and the two filariasis. PMID- 2545371 TI - Diagnostic and prognostic importance of tumor markers in lung cancer. PMID- 2545372 TI - Tumor markers for the diagnosis, prognosis, treatment and follow-up of gynaecological tumors. AB - It would be of benefit for the clinical relevance of tumor marker determination to be demonstrated, as a lot of markers are now in clinical use. Increased levels of carcinoembryonic antigen correlate with the stage of breast carcinoma. CA 15-3 should also be measured during follow-up of patients with this disease. The latest findings suggest a higher sensitivity and specificity of CA 15-3 than of CEA. The prognostic value and the usefulness of CEA measurement in screening seem to be poor. The measurement of CA 125 seems to be a reliable method for monitoring the presence and clinical behavior of ovarian cancer. It is suggested that invasive diagnostic procedures are not required in patients with normal marker levels. The management of chorion carcinoma can be determined as an ideal model in the range of marker application. Only in this disease does the marker HCG reach almost 100% sensitivity and specificity. The definition of response to chemotherapy and the appearance of relapse can be based on HCG measurement. PMID- 2545373 TI - Developmental expression of the chicken nerve growth factor receptor gene during brain morphogenesis. AB - Neural development proceeds in an ordered fashion in which a variety of genetic and epigenetic factors exert an influence at well defined times. Using a cloned chicken genomic fragment for the nerve growth factor (NGF) receptor, we have detected strong expression in chicken brain at early stages of embryonic development. Expression of the receptor gene was greatly diminished at birth. This pattern of NGF receptor mRNA level was observed in all cranial regions and was further correlated with the appearance and disappearance of cell surface receptors. The transient developmental expression of NGF receptors in chick brain and the requirement for receptors to mediate NGF's effects suggests that NGF may possess a broader range of actions during development of the nervous system. PMID- 2545374 TI - Alteration of benzodiazepine receptors in mouse cerebellum following methylazoxymethanol treatment during development. AB - The specific binding of [3H]flunitrazepam was studied to biochemically specify the morphological alterations induced in mouse cerebellum by a single injection of an antimitotic agent, methylazoxymethanol (MAM) performed at the beginning of the postnatal life. The MAM injection causes a general reduction of the benzodiazepine receptors in the adult mice which is particularly severe in mice having been injected the 1st day of postnatal life (so-called MAM0 mice) as compared to animals injected the 5th day (MAM5 mice): in MAM0 mice the benzodiazepine receptor is reduced to half of the control value. The affinity of the benzodiazepine towards its receptor was not affected and the topographic and biochemical action of MAM in the central nervous system was ascertained. Correlations could be made between the biochemical modifications and the morphological alterations otherwise described. PMID- 2545375 TI - Detection and identification of the plant lignans lariciresinol, isolariciresinol and secoisolariciresinol in human urine. AB - The mammalian lignans enterolactone and enterodiol are regular constituents of human urine and are excreted daily in mumol amounts. They are produced by metabolic action of intestinal bacteria from natural plant lignan precursors which are constituents of various food plants. However, natural plant lignans seem to occur very seldom in detectable amounts in human urine. The present investigation shows that only in 5% of the 150 diphenolic fractions extracted from the urine of women plant lignans other than the previously identified matairesinol could be found. The lignans lariciresinol, isolariciresinol and secoisolariciresinol were identified for the first time by comparison of their GC characteristics and mass spectra with the corresponding results of authentic synthesized reference compounds. Secoisolariciresinol is one natural precursor of the mammalian lignan enterodiol. Of the two other plant lignans, no animal or human metabolic products are known. The occurrence of chemically unchanged plant lignans in some cases in human urine could be a result of an insufficient metabolic capacity of intestinal bacteria, resulting in a decreased detoxification of phenolic plant products. PMID- 2545376 TI - Increased circulating concentration of atrial natriuretic factor in persons with pheochromocytomas. AB - To investigate the possible relationship of atrial natriuretic factor (ANF) to hypertension, we examined the circulating levels of ANF in 3 patients with pheochromocytomas before surgery, during increase of their blood pressure with surgical manipulation of their tumors, and after surgery when their blood pressures returned to normal. The circulating levels of ANF were increased 2-fold in patients with both extra-adrenal and intra-adrenal pheochromocytomas. In both the intra-adrenal and extra-adrenal patients their ANF levels increased further during surgical manipulation and returned to normal after surgical removal of their respective tumors. Each of these pheochromocytomas was examined and found to have atrial natriuretic receptors that were functional since ANF could enhance the guanylate cyclase - cyclic GMP system two-fold in these pheochromocytomas. We conclude that ANF circulates at higher concentrations in persons with pheochromocytomas and returns to normal with removal of the tumor. In addition, pheochromocytomas contain specific ANF receptors and ANF itself within these tumors. PMID- 2545377 TI - Effects of co-culture with vascular endothelial cells on the renin-like enzyme production in vascular smooth muscle cells. AB - Cultured human vascular smooth muscle cells (VSMC) produced an immunologically specific renin-like enzyme. The production of the renin-like enzyme was increased by 1 mM N6-O2'-dibutyryladenosine 3',5' cyclic monophosphoric acid sodium salt (db-cAMP). When co-cultured with human vascular endothelial cells (EC) and human skin fibroblast (FB), the basal production of the renin-like enzyme was increased. And the extent of the increase by db-cAMP was amplified by co-culture with EC, but not by the co-culture with FB. PMID- 2545378 TI - Lymphocyte basal cyclic AMP production predicts blood pressure. AB - Cyclic AMP (cAMP) serves as the second messenger for a variety of receptor systems. While much attention has been placed on the importance of receptor stimulated levels of cAMP accumulation, little attention has been given to the potential importance of the unstimulated basal levels. We measured both stimulated and unstimulated cAMP accumulation, as well as receptor number, in lymphocytes in 15 subjects and found relatively strong associations between basal cAMP accumulation and systolic (p = .004) and diastolic (p less than .001) blood pressure. Blood pressure was unrelated to either receptor number or adrenergically-stimulated levels of cAMP. These results underscore not only the complexity of the role of cAMP in the regulation of blood pressure but also the usefulness of peripheral blood cells as models of more central physiological regulation. PMID- 2545379 TI - Alpha 2-adrenoceptor regulation of parathyroid hormone function in the isolated perfused kidney. AB - We investigated physiological interactions between alpha 2-adrenoceptors and parathyroid hormone (PTH) in the isolated buffer-perfused kidney. PTH infusion (10nM) caused a rapid and significant rise in cAMP excretion which diminished despite continued hormone infusion. PTH also caused a delayed phosphaturia which peaked 20-30 minutes following the start of PTH infusion. alpha 2-Adrenoceptor stimulation with epinephrine (28nM) diminished PTH-stimulated cAMP accumulation by 68-71% (p less than 0.05) but had no effect on PTH-induced phosphaturia. None of the experimental interventions affected renal hemodynamics. In additional studies, we used lithium (1mM) as a marker of proximal tubular sodium transport. PTH caused a rapid rise in lithium excretion which was temporally distinct (maximum response in 0-10 minutes) from the phosphaturia. alpha 2-Adrenoceptor stimulation completely blocked the inhibitory effect of PTH on lithium reabsorption. These results suggest that alpha 2-adrenoceptors regulate the stimulatory effect of PTH at proximal tubular adenylate cyclase. However, alpha 2 adrenoceptors play no role in phosphate transport in this segment. alpha 2 Adrenoceptor stimulation reverses PTH-induced lithium excretion, suggesting physiological antagonism in the proximal tubule, most likely involving Na/H exchange. PMID- 2545380 TI - Alpha-adrenoceptor subtypes in blood vessels: physiology and pharmacology. AB - The existence of alpha 1- and alpha 2-adrenoceptors subtypes is now well established. Presynaptically, alpha 2-adrenergic autoreceptors modulate norepinephrine release. In vascular smooth muscle both alpha 1- and alpha 2 adrenoceptors exist postjunctionally, the alpha 1 subtype being innervated preferentially in most vascular beds. alpha 2-Adrenoceptors may be present on the smooth muscle cell as well as the endothelial cell where they may participate in the release of endothelium derived relaxing and constricting factors. Developing insights into the role of alpha-adrenoceptor subtypes in the cardiovascular system, and into the classification of subtypes of alpha 1- and alpha 2 adrenoceptors may be expected to have, if confirmed, significant therapeutic implications for drug discovery in the cardiovascular system. PMID- 2545381 TI - Alpha 2-adrenoceptor stimulation inhibits cellular cyclic AMP production in microdissected human glomeruli. AB - The physiological role of numerically predominant alpha 2-adrenoceptor in the kidney is still unknown. This study examined the effect of alpha 2-adrenoceptor stimulation on the production of cAMP from isolated human glomeruli. Unaffected portions of human kidneys which had been removed because of renal cell carcinoma were used for the study. Glomeruli were dissected manually under a stereo microscope. In these glomeruli, alpha 2-adrenoceptor stimulation with epinephrine in the presence of propranolol inhibited significantly parathyroid hormone dependent increases in cAMP production. This inhibitory effect of epinephrine was removed by adding a specific alpha 2-adrenoceptor antagonist, yohimbine, indicating that the inhibitory effect of epinephrine on cAMP formation was due to alpha 2-adrenoceptor stimulation. Thus, alpha 2-adrenoceptors are involved in the inhibition of cAMP production in human glomeruli. PMID- 2545382 TI - Interaction between alpha 2-adrenergic and NPY receptor pathways in human erythroleukemia cells. AB - Neuropeptide Y (NPY) appears to be a transmitter of the sympathetic nervous system, and its actions are similar to those of alpha 2-adrenergic receptor stimulation. In human erythroleukemia (HEL) cells, both NPY and epinephrine (acting through alpha 2-adrenergic receptors) inhibit adenylate cyclase and mobilize intracellular Ca++. We investigated possible interactions between NPY and epinephrine. In radioligand binding assays NPY did not alter antagonist or agonist binding to alpha 2-adrenergic receptors. NPY and epinephrine did not act synergistically to elevate intracellular Ca++. Neither agent alone, nor both together, affected the intracellular pH of HEL cells. Preincubation with NPY (like epinephrine) redistributed the alpha 2-adrenergic receptors away from the cell surface and into a sequestered pool. PMID- 2545383 TI - Functional relationship between platelet alpha 2-adrenoceptors and sympathetic nerve activity in man. AB - Functional relationships between platelet alpha 2-adrenoceptors and sympathetic nerve activity were studied in patients with essential hypertension (n = 23), primary aldosteronism (n = 10), and normal subjects (n = 12). The maximum number of binding sites (Bmax) and the dissociation constant (Kd) of 3H-yohimbine on platelet membranes and resting plasma norepinephrine (NE) were similar among these groups of subjects. In all subjects combined, Bmax values were correlated positively (r = 0.471, p less than 0.01) to resting plasma NE and negatively (r = -0.531, p less than 0.01) to the changes in plasma NE when subjects were moved from lying to standing. A 2-4 week treatment with guanabenz or bethanidine induced a parallel fall of both Bmax and plasma NE, whereas administration of reserpine caused a similar fall of plasma NE but Bmax remained unchanged. Kd values were not changed after any of these treatments. The results support the view that platelet alpha 2-adrenoceptor density is functionally regulated in parallel with sympathetic nerve activity rather than plasma NE per se. This hypothesis seems quite reasonable when it is extended to the presynaptic alpha 2 adrenoceptors; otherwise, the negative feedback function on NE release should be lessened and contradictory to its physiological role. PMID- 2545384 TI - Lymphocyte beta 2-adrenergic receptors in essential hypertension: studies in basal conditions and after dynamic exercise. AB - In order to study the density and response to dynamic exercise of lymphocyte beta 2-adrenoceptors in essential hypertension, 30 moderate essential hypertensives (12 men: 18 women, mean age 50.3 +/- 9.2 years) and 14 normotensive controls (8 men: 7 women, mean age 47 +/- 15 years) were studied. Beta 2-adrenoceptors density in basal conditions was higher in essential hypertensives than in controls (1950 +/- 625 vs 1278 +/- 356 binding sites per cell [bs/c], respectively, p less than 0.01) (mean +/- SD). After dynamic exercise beta 2 adrenoceptors density increased both in controls (to 1491 +/- 342 bs/c, p less than 0.01) and in hypertensives (to 2632 +/- 715 bs/c, p less than 0.001). However, the percentage of increase after the exercise was significantly higher in hypertensives than in controls (40.5 +/- 6.1 vs 20.2 +/- 6.5%, p less than 0.05). In summary, the higher density in basal conditions, as well as the exaggerated increase in the number of beta 2-adrenoceptors after dynamic exercise observed in essential hypertension, suggest an impaired regulation of these receptors in essential hypertension. PMID- 2545385 TI - Stimulation of norepinephrine release by peripheral presynaptic beta adrenoceptors. AB - Increases in plasma norepinephrine (NE) concentration induced by beta-agonist infusion have been taken as evidence for the existence of peripheral presynaptic beta-adrenoceptors, facilitating NE release. Concomitant hemodynamic changes, however, could invoke reflex mechanisms and thus hamper the interpretation of the results. We studied the presence of peripheral presynaptic beta-adrenoceptors by giving intra-arterial infusions of EPI in the forearm before and during uptake inhibition, and the effects of systemic infusions of very low-dose epinephrine (EPI) and isoproterenol (ISO) on hemodynamics and arterial and venous plasma NE concentrations. An i.a. infusion of EPI, 0.1 ng.kg-1.min-1, increased the net overflow of NE from 0.3 +/- 1.0 to 5.4 +/- 1.8 pmol.min-1 (M +/- SEM, p less than 0.05) and raised the calculated arterial plasma EPI concentration to 0.92 +/- 0.22 nmol.l-1. The uptake inhibition increased the net overflow of NE to 15.4 +/- 2.4 pmol.min-1 and during addition of EPI, 0.1 ng.kg-1.min-1, it further increased to 23.0 +/- 4.1 pmol.min-1 (p less than 0.05), while it decreased during addition of an effective beta-blocking dose of propranolol, 50 ng.kg-1.min 1 i.a., to 18.2 +/- 4.1 pmol.min-1 (n.s.). Systemic infusions of EPI and ISO increased blood pressure, heart rate, and forearm blood flow before significant changes in arterial or venous plasma NE concentrations were found. It is concluded that the increases in net NE overflow from the forearm during local EPI infusions indicate the presence of peripheral presynaptic beta-adrenoceptors which are responsive to physiologic concentrations of EPI. Systemic infusions of EPI or ISO elicit hemodynamic effects, and thus may invoke reflex mechanisms, before an influence on arterial or venous plasma NE concentration is found. PMID- 2545386 TI - Infusions of adrenaline stimulate noradrenergic transmission in man, but adrenaline is not released later as a co-transmitter. AB - A pre-junctional beta-adrenoceptor facilitating noradrenergic transmission may be affected by elevations in circulating adrenaline, and represent a mechanism whereby mental stress might predispose to hypertension. The proposition that circulating adrenaline can be taken up into noradrenergic nerve terminals and later co-released with noradrenaline in response to an appropriate stimulus was examined. The effects of upright posture, before and after a 60 min adrenaline infusion were as follows: (1) Changes in heart rate and diastolic blood pressure were not augmented. (2) Plasma noradrenaline response was not increased. (3) Plasma adrenaline basally and in response to posture remained low. (4) Radiolabelled adrenaline disappeared from the circulation on cessation of infusion, and did not reappear in response to posture or isometric exercise. While adrenaline infusion again increased noradrenaline, we found no evidence of subsequent adrenaline release in response to noradrenergic stimulation. These studies do not exclude adrenaline uptake and re-release in amounts sufficient to stimulate the prejunctional beta-adrenoceptor, but insufficient to reach the circulation after local metabolism. PMID- 2545387 TI - Diagnosis of salivary gland disease using ultrasound and sialography: a comparison. AB - Thirty-one patients with a suspected inflammatory or neoplastic lesion of a major salivary gland were investigated with ultrasound, and in 27 of them sialography was carried out in addition. The results of the diagnostic procedures were compared. Lesions of the gland parenchyma and intra and extra-glandular duct ectasia were demonstrated by ultrasound. Salivary stones were demonstrated by this method and ultrasound allowed assessment of periglandular structures. Sialography is indispensable for detailed assessment of the salivary ducts. The present report shows that ultrasound is superior to sialography in the assessment of salivary gland disease. The investigation is non-invasive, cheap, and requires neither contrast nor radiation and can be repeated at will for follow-up. PMID- 2545388 TI - Parapharyngeal tumours: a review of 23 cases. AB - The anatomy of the parapharyngeal space is described and experience with 23 patients with parapharyngeal tumours seen over a 10-year period is reported. Despite reaching a large size, these tumours may be relatively asymptomatic, or be discovered incidently. CT scan, MRI, arteriography and fine needle aspiration are useful in establishing a diagnosis. The most common histological type (13/23 patients) was pleomorphic adenoma and these tumours were managed by transoral removal if small, or a combined transoral-external approach if large. No patient required a mandibulotomy for access and no benign tumour recurred. The outcome was poor in the 5 patients with malignant tumours and a more radical approach, midline mandibulotomy, should be considered if malignancy is suspected. PMID- 2545389 TI - Magnetic resonance with gadolinium DTPA for the investigation of temporal bone tumours. AB - The membranous labyrinth and 7th and 8th cranial nerves are well shown by magnetic resonance scanning but the lack of bone detail makes MRI secondary to CT for the investigation of lesions of the petrous temporal bone. Twenty-two intrinsic temporal bone tumours were examined by magnetic resonance in this study. Seven glomus tumours, one facial neuroma and one carcinoid tumor received intravenous gadolinium DTPA. All showed signal enhancement although the practical value of this enhancement was variable. The ability of MRI to demonstrate the jugular bulb separate from a glomus tumour confined to the middle ear, and signal enhancement of another middle ear tumour after partial removal are described, as well as the tissue characterization for discriminating between cholesteatoma and cholesterol granuloma for mass lesions in the petrous apex. PMID- 2545390 TI - Paradoxical effects of endothelin on cardiovascular noradrenergic neurotransmission. AB - 1. Pre- and postjunctional effects of endothelin (1-10 nmol/L) have been studied in the rabbit isolated ear artery and in rat isolated atria. 2. Endothelin produced concentration-dependent increases in arterial perfusion pressure, and had positive chronotropic and inotropic effects in rat atria. 3. Vasoconstrictor responses of the arteries to sympathetic nerve stimulation were reduced by 1 nmol/L endothelin and abolished by 10 nmol/L endothelin. In rat atria, the chronotropic responses to nerve stimulation were markedly reduced by 10 nmol/L endothelin. 4. In rabbit ear artery, vasoconstrictor responses to noradrenaline were enhanced by 1 nmol/L endothelin, but were reduced by 10 nmol/L endothelin. In rat atria, endothelin reduced the chronotropic response to isoprenaline. 5. Endothelin (10 nmol/L) increased the stimulation-induced release of radioactivity in arteries and atria labelled with [3H]-noradrenaline by 91% and 23%, respectively. 6. The pre- and postjunctional effects of endothelin persisted in both arterial and atrial preparations for at least 30 min after its removal. PMID- 2545391 TI - Endothelin receptors in rat adrenal gland visualized by quantitative autoradiography. AB - 1. The radioligand [125I]-endothelin was used to map receptors for endothelin in rat adrenal gland using in vitro autoradiography and computerized densitometry. 2. In the adrenal, a high density of binding was found in the adrenal medulla (binding affinity constant 0.18 +/- 0.11 X 10(9)M-1) and zona glomerulosa (binding affinity constant 0.18 +/- 0.07 X 10(9)M-1). Binding was low to undetectable in the zona fasciculata and zona reticularis. Unrelated peptides did not displace endothelin. 3. These results provide evidence of endothelin receptor distribution in adrenal gland and suggest that endothelin might exert multiple actions in the adrenal gland on catecholamine and aldosterone biosynthesis and secretion. PMID- 2545392 TI - Nifedipine blocks ACTH and cortisol release in man. AB - 1. The present study investigated the effect of prior administration of nifedipine on AVP-induced ACTH release in seven normal volunteers. Three protocols were used: 20 mg oral nifedipine; 0.14 pressor units intramuscular (i.m.) per kg bodyweight aqueous AVP; oral nifedipine plus i.m. AVP 90 min later. Plasma ACTH and cortisol were measured at intervals for 2.5 h during each test. 2. The mean peak plasma ACTH and cortisol levels and the mean peak changes from basal in these levels were significantly lower in the nifedipine/AVP test than in the AVP alone test. The integrated area under the cortisol time curve was significantly lower for the nifedipine/AVP test than that for the AVP test alone. Nifedipine alone caused no changes in ACTH or cortisol. 3. Acute administration of oral nifedipine caused an inhibition of AVP-stimulated ACTH and cortisol release in normal humans. This effect may be due to blockade of plasma membrane calcium channels normally activated during AVP stimulation of pituitary corticotrophs. PMID- 2545393 TI - Potentiation of fenfluramine-induced ACTH release in man by naloxone. AB - 1. This study set out to compare the effect of the combination of fenfluramine (Fen) and naloxone (Nal) on ACTH and cortisol release with each of these agents alone in six normal volunteers. Three protocols were used: 1.5 mg/kg bodyweight oral Fen; 125 micrograms/kg bodyweight i.v. Nal; oral Fen plus i.v. Nal 135 min later. Plasma ACTH and cortisol were measured at intervals of 285 min over each test. 2. The mean peak levels and mean peak changes from basal in plasma ACTH and cortisol were significantly greater in the combination test (Fen/Nal) than in either the Fen alone or the Nal alone tests. Fen and Nal given together cause a synergistic release of ACTH and cortisol as determined by the areas under the ACTH-time and cortisol-time curves for Fen/Nal compared to the sums of Fen alone plus Nal alone curves for ACTH and cortisol. 3. We conclude that Fen and Nal acting synergistically work through different and potentiating mechanisms at the hypothalamic, central nervous system and/or pituitary levels to cause ACTH release. PMID- 2545394 TI - Angiotensin converting enzyme in the rat heart: studies of its inhibition in vitro and ex vivo. AB - 1. The pharmacokinetics of angiotensin converting enzyme (ACE) inhibition in rat heart and lung was evaluated in vitro and ex vivo. 2. Radioinhibitor [125I]-351A binding displacement was used to assess the relative potency of six ACE inhibitors (CI906, CGS14831, S9780, 351A, MK521, SQ27519) in rat heart and lung homogenates, and estimate equilibrium association constant (Ka). 3. Following oral administration of 0.3 mg/kg of Quinapril (CI928) specific binding of [125I] 351A to ACE was measured in rat heart. 4. Ka for binding to ACE of each inhibitor was significantly higher in right and left atrium than in lung (P less than 0.05) or the right and left ventricle (P less than 0.005). These differences did not affect the degree or time course of inhibition in vivo in the rat myocardial ACE following Quinapril treatment. 5. Rank order of potency of the ACE inhibitors tested was CI906 = CGS14831 greater than S9780 greater than 351A greater than MK521 greater than SQ27519 PMID- 2545395 TI - Effect of enalapril on handling of a sodium chloride load by genetically hypertensive and normotensive rats. AB - 1. Enalapril was given in the drinking water (300 mg/L) for 4.5 days to normotensive (N) and genetically hypertensive (GH) rats on zero sodium intake. An intraperitoneal NaCl load was given 12 h after enalapril was started. 2. Enalapril did not increase the maximum rate of sodium excretion, but caused the rats to excrete more than the load in the first 24 h and then to have a slow fall in body sodium while on a sodium-free diet. 3. In terms of the Strauss et al. (1958) concept of body sodium, enalapril appears to lower the basal level. However, in addition it causes a slow leak of sodium which becomes apparent when sodium intake is very low. PMID- 2545396 TI - Mouse astrocytes possess specific ANP receptors which are linked to cGMP production. AB - 1. Receptors for atrial natriuretic peptide (ANP) have been identified on mouse astrocytes in primary culture, and have similar characteristics to those found on previously recognized ANP-target tissues. 2. Scatchard analysis revealed one class of high affinity receptors with a Kd of 0.32 nmol/L. The IC50 for specific binding was 0.5 nmol/L. 3. Ligand binding resulted in stimulation of guanylate cyclase. 4. Under reducing conditions, the covalently cross-linked receptor-ANP complex migrated on SDS-polyacrylamide gels as a single band with an apparent molecular weight of 66 kDa. 5. Although the physiological relevance of our observations remains to be determined, these data document that cultured mouse astrocytes contain specific high-affinity ANP receptors which are linked to the production of cGMP. PMID- 2545397 TI - Ultrastructural study of nuclear evagination in neoplastic cells of a cerebral primitive neuroectodermal tumor. AB - Evaginations of the nuclear envelope containing nuclear material were observed in neoplastic cells of a cerebral primitive neuroectodermal tumor. The possible sequential development of nuclear evaginations included: 1) formation of a segmental naked inner membrane of the nuclear envelope due to loss of the underlying marginal heterochromatin, 2) elevation of the overlying outer nuclear membrane with focal expansion of the perinuclear cistern, 3) outpouching of the inner nuclear membrane forming a small bleb, and 4) protrusion of electron-lucent nuclear material into the bleb forming a nuclear evagination. Some nuclear evaginations were isolated in the cytoplasm and some protruded into the intercellular space. The inner nuclear membrane of the nuclear evagination was devoid of the marginal heterochromatin. Nuclear pores and rupture of membranes with release of the contents of nuclear evaginations into the cytoplasm were not observed. Instead, the nuclear evaginations isolated from the cytoplasm often exhibited degenerative changes with disintegration of their contents and inner nuclear membrane. It is suggested that the nuclear evaginations of the neoplastic cells in the present study may represent a means to eliminate excess nuclear material. PMID- 2545398 TI - A clinical-pathologic-biochemical study of the membrane surrounding loosened and nonloosened total hip arthroplasties. AB - The clinical and roentgenologic data from 31 excised components from 19 revision arthroplasty cases were correlated with the histology and biochemistry of the membrane at the bone-cement or bone-prosthesis interface. Twenty-seven components were cemented and four were uncemented. Twenty-four implants were clinically and roentgenologically loose, one was possibly loose, and six were well fixed. Loose components, whether cemented or not, demonstrated statistically higher prostaglandin E2 levels in the surrounding membrane compared to the nonloose group. Collagenase and M-collagenase levels were absent or insignificantly low in all specimens; no detectable interleukin 1 beta was found. This suggests that prostaglandin E2 may be associated with the bone lysis associated with prosthesis loosening. PMID- 2545399 TI - Establishment of a cell line producing bone morphogenetic protein from a human osteosarcoma. AB - A human osteosarcoma cell line was established from a biopsy specimen from a 13 year-old girl. The osteosarcoma tissue was maintained in athymic nude mice (Balb C nu/nu) by serial transplantation for three years. The tumor was excised from a host mouse and digested with collagenase. The isolated cells were cultured by 98 passages in 14 months, and clones of osteosarcoma cells were obtained by limiting dilution. A clone named human osteosarcoma cell 6 (H-OS-6) that showed the osteoblastic phenotypes of productions of bone morphogenetic protein (BMP) and alkaline phosphatase and a response to human parathyroid hormone (h-PTH 1-34) was selected. The morphology of its chromosomes indicated its human origin. This human osteosarcoma cell line is unique in producing BMP under in vitro conditions. PMID- 2545401 TI - [A case of meningoencephalitis caused by persistent Epstein-Barr (E-B) virus infection]. AB - A 44-year-old man had several episodes of tingling sensation in his lower extremities, head and face over 1 year and presented mental disturbance 2 months prior to admission. He additionally suffered from dysuria and acute onset of gait disturbance 18 days before admission. Physical and neurological examination revealed marked splenomegaly, stupor, abnormal behavior, spastic tetraparesis and sphincter disturbances, and meningoencephalitis was suspected. There was lymphocytosis in peripheral blood, some of which showed atypical morphology. CSF examination revealed increased protein content and mononuclear pleocytosis. The titers of antibodies against E-B virus were elevated as follows; VCA IgM (X320), VCA IgG (X20,480), EA (X10,240) and EBNA (X10) in serum, and VCA IgM (X4), VCA IgG (X160), EA (X40) and EBNA (less than 1) in CSF. CT and MRI examination revealed ring enhanced lesion adjacent to left lateral ventricle with surrounding diffuse edema. Administration of intravenous glycerol and oral prednisolone induced substantial improvement in sensory, mental and sphincter disturbances and brain CT findings but not in splenomegaly and pyramidal tract signs. Antibody titer against VCA IgM also decreased. After withdrawal of prednisolone, CSF and CT findings worsened, and the antibody titer became elevated again, and CSF and CT findings improved by readministration of prednisolone. From the above signs and laboratory data, especially continuous elevation of antibody titer against E B VCA IgM for as long as 1 year, this case was considered to be a meningoencephalitis caused by persistent E-B virus infection. PMID- 2545400 TI - [A case of recurrent Epstein-Barr virus meningitis]. AB - A 17-year-old male was admitted to our hospital complaining of fever, headache and nausea. On physical examinations cervical and inguinal lymphadenopathy and hepatosplenomegaly were noted. Neurological examination revealed meningeal signs. Blood examination showed slightly increased lymphocytes and atypical lymphocytes (1-4%), increased ESR, CRP (2+), slightly elevated EBV VCA IgG (X20), and normal EBV VCA IgM (less than X 10). Initial pressure of CSF was 195mmH2O, cells 46/3mm3, protein 50mg/dl. Slowing of back ground activity of EEG such as theta and delta wave was noted. CT scan revealed normal. During the course EBV VCA IgG elevated to X160, EBV VCA IgM elevated to X20. Slight respiratory disturbance, photophobia, and dysosmia were noted. One month later, clinical symptoms and laboratory data improved, the patient was discharged. Forty days after the discharge, headache was excerbated and increased CSF protein (100mg/dl) and pleocytosis (33/3mm3) were noted. The patient was readmitted to the hospital. After 2 weeks in the hospital, symptoms were diminished and laboratory findings revealed normal. During 2 years before the third admission he was asymptomatic and could enjoy the college life. At age 19, he was admitted to our hospital complaining of fever, headache and nausea. Neurologically slight meningeal irritation was noted. Blood examination revealed 1% atypical lymphocytes. EBV VCA IgG was elevated (X320) and EBNA was X80. CSF protein was slightly increased (52mg/dl). During the clinical course CSF protein was elevated to 105mg/dl and cell count to 502/3mm3 (N: L = 27: 409, with 66 atypical lymphocytes). Persistent hiccup was noted. After 50 days CSF findings were improved and he was discharged.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545402 TI - Gastric pertechnetate contribution to blood pool images. AB - During a cardiac blood pool study (Tc-99m erythrocytes), there was an apparent "cold" area in the left lobe of the liver. At one hour, the prior void appeared to have filled in, suggesting an intrahepatic hemangioma. However, the changing pattern of radioactivity, and its shape, indicated that this actually represented Tc-99m pertechnetate in the gastric wall and stomach cavity. In a second case, the interval between intravenous injection of stannous pyrophosphate and Tc-99m pertechnetate was prolonged. Changing activity, again in the gastric wall and cavity, obscured part of the liver. This cleared with time, allowing hepatic evaluation. Thus, gastric pertechnetate can contribute to blood pool images of the heart or liver and multiple images may be needed to separate this component from nearby tissues. PMID- 2545403 TI - Caliceal outpouching: diagnosis by technetium-99m DTPA and DMSA renal scintigraphy. PMID- 2545404 TI - Role of excitatory amino acids in brain injury caused by hypoxia-ischemia, status epilepticus, and hypoglycemia. AB - Glutamate, the major excitatory neurotransmitter of the brain, mediates neuronal injury from hypoxia-ischemia, hypoglycemia, and status epilepticus. Drugs that block glutamate receptors, particularly the N-methyl-D-aspartate (NMDA) receptor, protect neurons from these insults. Noncompetitive antagonists of NMDA receptors have the potential to prevent perinatal neurologic morbidity. PMID- 2545406 TI - Effects of etodolac on human chondrocytes cultivated in three dimensional culture. AB - Human chondrocytes in three-dimensional culture were incubated for up to 20 days in the presence of etodolac, a nonsteroidal anti-inflammatory drug which penetrates readily into the synovial fluid. Even at an etodolac concentration of 80 micrograms/ml, DNA synthesis, proteoglycan synthesis, and type-II collagen synthesis were unchanged. Collagenase production was also unaffected by etodolac (60 micrograms/ml). In contrast, prostaglandin E2 production was reduced by 84% in the presence of 60 micrograms/ml of etodolac. The 80 micrograms/ml concentration is 5 times that found in the serum of subjects treated with 200 mg of etodolac twice a day for 6 days, and 33 times the concentration in synovial fluid of arthritic patients treated with etodolac 200 mg twice a day for 7 days. These in vitro results indicate that anti-inflammatory levels of etodolac may not damage articular cartilage in vivo. PMID- 2545405 TI - Supravital dithizone staining in the isolation of human and rat pancreatic islets. AB - Dithizone, a zinc chelating agent, is known to selectively stain the islets of Langerhans in the pancreas. In the present study, we have used this stain to aid the identification of islets in material obtained by collagenase digestion of human pancreas. Islets were shown to rapidly and reversibly stain red on incubation with dithizone solution. Tissue selected on the basis of dithizone staining was shown to contain insulin-positive cells and to accumulate insulin in the medium during a subsequent period in tissue culture. Experiments with rat islets indicated that the dithizone treatment had no effect on insulin release in tissue culture, on acute responses to stimulatory glucose concentrations or on the insulin content of cells. These results suggest that dithizone staining can assist in the identification of islets from the human pancreas and may prove to be a useful tool in developing techniques for the large scale isolation of functionally intact human islets. PMID- 2545408 TI - [Peripheral rheumatoid neuropathy. Description of a clinical case]. AB - After describing peripheral rheumatoid neuropathy and pointing out its increased frequency after the advent of steroid therapy, the Authors describe a case brought to their attention at the Rheumatology Division of the Ospedale San Camillo De Lellis. PMID- 2545407 TI - Comparative effects of omega-3 fatty acids in men and women. PMID- 2545409 TI - Sodium bicarbonate cotransport in cultured pigmented ciliary epithelial cells. AB - Uptake of 22Na+ was studied in cultured bovine pigmented ciliary epithelial cells (PE) in HCO3-containing media. Two components of Na+-uptake were stimulated by intracellular acidification (NH4+-prepulse): One was amiloride-sensitive, the other DIDS-sensitive. The amiloride-sensitive component of Na+-uptake probably represents Na+/H+-exchange, which has previously been characterized in PE. The second, DIDS-sensitive component stimulated by intracellular acidification, was Cl- and HCO3--dependent. We conclude that a stilbene-sensitive, Cl--dependent Na+ HCO3--cotransport is present in PE. This transport could play an important role in aqueous humor formation. PMID- 2545410 TI - Lacrimal fluid and electrolyte secretion: a review. AB - Lacrimal gland fluid is an important component of the precorneal tear film. The rate of lacrimal gland fluid secretion is controlled primarily by parasympathetic innervation, and it is, apparently, modulated by sympathetic innervation. Lacrimal gland fluid is produced in two stages, secretion of a primary fluid which resembles an isotonic ultrafiltrate of plasma in the acinus-early intercalated duct region, and secretion of a KCl-rich fluid in subsequent ductal elements. Little is known about the electrolyte transport mechanisms of the ductal epithelia. Recent work using a variety of techniques, including tracer flux measurements, intracellular electrical recording, intracellular ion activity measurements, patch clamping, and analytical subcellular fractionation, supports a model for transcellular Cl-secretion in the acinus which involves Cl--selective channels in the apical plasma membrane and an array of Na+/H+ antiporters, Cl /HCO3-antiporters, K+ channels, and Na,K-ATPase in the basal-lateral plasma membrane. PMID- 2545411 TI - Signal transduction and control of lacrimal gland protein secretion: a review. AB - Proteins in lacrimal gland fluid are secreted primarily by the acinar cells. Secretory proteins are synthesized in the endoplasmic reticulum, modified in the Golgi apparatus, stored in secretory granules, and released upon a change in the cellular level of second messenger. The second messenger level is controlled by a process termed signal transduction. Agonists, primarily neurotransmitters in the lacrimal gland, bind to receptors in the basolateral membrane of secretory cells. This interaction activates enzymes in the membrane that cause production of second messengers. It has been hypothesized that second messengers stimulate secretion by activating specific protein kinases to phosphorylate proteins important for secretion. In the lacrimal gland, cholinergic agonists stimulate protein secretion. They act by activating phospholipase C to break down phosphatidylinositol bisphosphate into 1,4,5-inositol trisphosphate (1,4,5-IP3) and diacylglycerol (DAG). 1,4,5-IP3 causes release of Ca2+ from intracellular stores. This Ca2+, perhaps in conjunction with calmodulin, activates specific protein kinases that may be involved in secretion. DAG activates protein kinase C which stimulates protein secretion. alpha 1-Adrenergic agonists also stimulate lacrimal gland protein secretion. These agonists use a pathway that is separate from that utilized by cholinergic agonists and vasoactive intestinal peptide (VIP). The specific pathway has not been identified but may be DAG and protein kinase C. VIP, beta-adrenergic agonists, alpha-melanocyte stimulating hormone, and adrenocorticotropic hormone are lacrimal gland secretagogues. They activate adenylate cyclase to produce cAMP. cAMP stimulates protein kinase A, which perhaps causes protein secretion. Thus, three separate cellular pathways stimulate lacrimal gland protein secretion. Cholinergic agonists and VIP also stimulate lacrimal gland fluid secretion, and the same signal transduction pathways utilized by these agonists to stimulate protein secretion are most likely used for electrolyte and water secretion. PMID- 2545412 TI - Disorders of phagocyte killing and digestion (CGD, G-6-PD and myeloperoxidase deficiencies). PMID- 2545413 TI - Herpesvirus infections in the immunocompromised host. Clinical spectrum, diagnosis, management and prophylaxis. PMID- 2545414 TI - Severe eczema herpeticum is associated with prolonged depression of cell-mediated immunity to herpes simplex virus. AB - Nine patients with eczema herpeticum (EH) and 2 with atopic eczema and recurrent cold sores were investigated for cell-mediated immunity (CMI) and antibodies (Ab) to herpes simplex virus (HSV). All patients had Ab to HSV and a high lymphoproliferation response to concanavalin A and all but 3 showed lymphoproliferation to HSV antigens. These 3 patients had very severe EH and absent CMI to HSV for many months after clinical recovery despite normal Ab and concanavalin A responses. In 2 of them, depletion of CD8+ T cells by panning restored the lymphoproliferative response to HSV. Thus their absent CMI may be due to suppressor cell function rather than lack of responsive cells. PMID- 2545415 TI - Immune response to papillomavirus infections. PMID- 2545416 TI - Human papilloma virus infection and its relationship to skin cancer in a group of renal allograft recipients. PMID- 2545417 TI - What's new in the treatment of difficult infectious diseases? PMID- 2545418 TI - Proviral position effects: possible probes for genes that suppress transcription. AB - Rous sarcoma virus, an oncogenic avian retrovirus, readily causes morphological transformation of chick cells, but in infected rat cells transformation is rare because proviral transcription is inefficient. This constraint is not due to a lack of positive transcriptional factors, or an excess of negative ones, but reflects the site of proviral integration in rat cell DNA. In most sites the provirus is almost invariably silent, in others it is correspondingly active, whilst in a third category expression fluctuates in concert with transitions in chromatin structure. Transcriptional fluctuations are mediated both by flanking cell DNA in cis and by trans-acting cell genes, suggesting that proviral position effects are sensors for genes that down-regulate transcription, perhaps by determining chromatin configuration. We have tried to identify such genes by gene transfer, karyology and insertional mutagenesis. The variable success of these three approaches indicates that the transcriptional down-regulator(s) need act only transiently. This is consistent with a function that operates in ontogeny or differentiation to down-regulate genes whose silence is then perpetuated by other means. The loss of such functions may predispose to neoplasia. PMID- 2545419 TI - A genetic basis for tumour suppression. AB - The technique of somatic cell hybridization has established the phenomenon of tumour suppression and provided evidence for a genetic basis for suppression. Further refinements aimed at eventually identifying 'tumour suppressor' genes include the use of monochromosome transfer via microcell hybridization. The application of this technique to the study of tumour suppression in tumorigenic HeLa cell x fibroblast hybrids, Wilms' tumour, retinoblastoma and osteosarcoma cells is described. The issue of whether tumour suppression involves a direct effect on expression of activated oncogenes is discussed. Transformation of normal human cells in culture by activated cellular oncogenes is an extremely rare event. This may be due to a relatively greater genomic stability of human cells compared to rodent cells. We describe the use of a spontaneously immortalized human keratinocyte cell line, HaCaT, for studies of the effects of introduction of activated c-Ha-ras oncogene into these cells, with particular reference to tumorigenic conversion. PMID- 2545421 TI - Transgenic mice and host cell mutants resistant to transformation as model systems for identifying multiple components in oncogenesis. AB - Tumorigenesis appears to be a multistep process involving mutations of conventional, dominantly acting proto-oncogenes, mutations of other genes that may act in a recessive manner, and interactions (or a lack of interactions) between the products of mutant and wild-type genes. Our laboratory is using a few well-established, dominant oncogenes to pose experimental questions that could lead to a better understanding of the more elusive genetic interactions which occur during tumour development. Two such situations are described: (1) We have created a line of transgenic mice that carry the int-1 proto-oncogene under the control of the enhancer element in the mouse mammary tumour virus long terminal repeat. Such mice express the transgene in mammary glands, salivary glands and male reproductive tract; mammary glands from both male and female animals are grossly hyperplastic, yet tumours arise rarely in the males and sporadically in the females (80% of female mice have one or a few tumours by six months of age). Thus expression of int-1 in these mice appears to place a large number of mammary cells at risk for secondary events that lead to carcinogenesis, providing a provocative experimental context for identifying such secondary events. (2) We have isolated a rat cell line that lacks most of the characteristics of transformed cells, despite the expression of two wild-type copies of the v-src gene of Rous sarcoma virus. This line harbours what appears to be a dominant mutation in an unidentified gene that renders the cell resistant to transformation by v-src and several other oncogenes. Isolation of the mutant gene responsible for suppressing transformation in this line should provide new insights into the interactions between oncoproteins and other cellular proteins. PMID- 2545420 TI - Molecular basis for the regulation of cell fate by the lethal (2) giant larvae tumour suppressor gene of Drosophila melanogaster. AB - Tumour suppressor genes act as recessive determinants of cancer. Their function is required for normal cell growth and differentiation during development. When both alleles of these developmental genes are inactivated, cell growth becomes unrestricted. In Drosophila, a series of genes have been identified which when mutated produce tissue-specific tumours. Of these the lethal(2)giant larvae (l(2)gl) gene is the best studied. Homozygous l(2)gl mutations cause the development of malignant tumours in the brain and the imaginal discs. Genomic DNA from the l(2)gl locus has been cloned, introduced back into l(2)gl mutant animals by P-element-mediated transformation and shown to restore normal development. The nucleotide sequence of the l(2)gl gene (13.1 kb) has been determined, as well as the sequences of the two classes of transcripts. These transcripts encode two polypeptides of 127 kDa and 78 kDa, respectively. Both proteins have been immunologically identified. Analyses of the spatial distribution of both l(2)gl transcripts and proteins revealed that during early embryogenesis the l(2)gl gene is uniformly expressed in all cells and tissues. In late embryos, the l(2)gl expression becomes gradually restricted to tissues presenting no morphological or neoplastic alteration in the mutant animals. Further mosaic experiments pointed out that the critical period for the establishment of tumorigenesis is limited to early embryogenesis at a time when the l(2)gl expression is most intense in all cells. PMID- 2545423 TI - Genetics of polyposis and colorectal cancer. PMID- 2545422 TI - Reversion of tumorigenicity in an EBV-converted Burkitt's lymphoma line. AB - One of five Epstein-Barr virus (EBV)-converted sublines of an EBV-negative Burkitt's lymphoma line (BL-41) was identified as a non-tumorigenic phenotypic revertant with low clonability, comparable to that of an EBV-transformed lymphoblastoid cell line (IARC-171) derived from the same patient. This revertant subline (BL-41/95) also showed the most LCL-like phenotype of the five convertants tested. It is suggested that reversion was due to the phenotypic shift of the cell from a 'window' of cell ontogeny that contains virgin B cells and memory B cells to the stage of the activated immunoblast. Constitutive activation of the c-myc gene by translocation to an immunoglobulin locus continues to drive proliferation of B cells in vivo even after they have undergone a programmed switch to a basically resting (virgin or memory cell) phenotype. An activated immunoblast invariably expresses c-myc. It is also suggested that proliferation of the activated immunoblast is regulated by negative host controls that prevent clonal overexpansion and keep the B cell pool constant. The sensitivity of the immunoblast to this control overrides the 'forward-driving' force of both EBV and the activated myc gene. PMID- 2545424 TI - Technetium pertechnetate esophageal imaging for detection of Barrett's esophagus. AB - This study was designed to evaluate the utility of 99mTc pertechnetate esophageal scintigraphy for identifying Barrett's esophagus. Seventeen patients with Barrett's esophagus and seven patients with reflux esophagitis were studied. Eight of 17 patients with Barrett's esophagus had a positive image (sensitivity 47%). In contrast, none of the seven patients with esophagitis had a positive image (specificity 100%). Pentagastrin did not have a significant effect on the sensitivity. There was no correlation between the type of Barrett's epithelium and the sensitivity of the imaging results. However, the test is more frequently positive in those patients with more extensive disease. Our study indicates that technetium pertechnetate imaging should not be used as a screening test for the detection of Barrett's esophagus in patients with symptoms of gastroesophageal reflux, as the negative predictive value of the test is limited. PMID- 2545425 TI - Effects of pectin on fatty acid and glucose absorption and on thickness of unstirred water layer in rat and human intestine. AB - Effects of pectin, a soluble dietary fiber, on fatty acid and glucose absorption were studied in vivo in rats and humans by perfusing the intestine with linoleic acid and glucose solutions with and without pectin. Linoleic acid and glucose absorption decreased with increasing concentrations of pectin. The reduction in linoleic acid absorption was not caused by binding of linoleic acids by pectin or impaired micelle formation due to binding of bile acid by pectin. The unstirred water layer expanded with increasing concentrations of pectin. These results suggest that enlargement of the unstirred water layer is closely associated with the reduction of absorption of fatty acid and glucose ingested with pectin. PMID- 2545426 TI - Hilar cholangiocarcinoma associated with clonorchiasis. AB - Pathologic and epidemiologic evidence support the relationship between the liver fluke Clonorchis sinensis and the development of bile duct cancer. We report here a case of cholangiocarcinoma in a Laotian immigrant originally diagnosed with clonorchiasis. This case demonstrates the importance of early recognition and treatment of this disease to prevent subsequent morbidity and mortality. Diagnosis and treatment of clonorchiasis as well as the possible modes of cancer induction are discussed. PMID- 2545427 TI - Proceedings of a workshop entitled "Neuromuscular function and dysfunction of the gastrointestinal tract in aging". PMID- 2545428 TI - [The effect of serotonin on the desensitization of GABA-receptors in mollusk neurons]. PMID- 2545429 TI - [A quantitative change in topoisomerase type I in the cell cycle of Chinese hamster fibroblasts]. PMID- 2545430 TI - [Ca 2+-dependent capture of small negatively charged liposomes by large unilamellar liposomes from phosphatidylcholine]. PMID- 2545431 TI - [Functional analysis of 5'-nontranslatable region of poliovirus RNA]. PMID- 2545432 TI - [Regulation of serotoninergic transmission in the basal ganglia]. PMID- 2545433 TI - [Neurotensin/dopamine interactions]. PMID- 2545434 TI - [Toxophilic effects of morphinomimetic substances. Dopamine/enkephalin interactions in the ventral tegmental area and in the nucleus accumbens]. PMID- 2545435 TI - Clinicopathologic analysis of endoscopic ultrasonograms in pancreatic mass lesions. AB - Endoscopic ultrasonography (EUS) was performed in 40 patients with pancreatic masses who subsequently underwent surgical resection, and we compared the ultrasonograms with the histopathologic findings. Ductal adenocarcinoma: The cephalad margin between the tumor and pancreatic tissue was distinct, while the caudal margin was blurred because the severe fibrosis accompanying secondary pancreatitis, and the contour of the tumor was irregular. The internal echo pattern was hypoechoic, with an uneven central echogenic portion corresponding to irregularly arranged carcinomatous canaliculi or coagulation necrosis. Benign islet cell tumor: Both the cephalad and caudal margins were distinct, the contour was smooth and the internal echo pattern was hypoechoic, with a homogeneous central echogenic portion corresponding to regularly arranged alveoli. Pseudotumorous pancreatitis: The caudal margin was indistinct, the contour was smooth, and the internal echo pattern was homogeneously hypoechoic, with deep attenuation caused by dense fibrosis. On the basis of these results, it is believed that EUS with its high resolution is useful in the differential diagnosis of pancreatic mass lesions. PMID- 2545436 TI - Quartz exposure, retention, and early silicosis in sheep. AB - The purposes of this study were (1) to investigate the chronology of events in cellular and biochemical changes thought to be important in the development of silicosis, (2) to relate these to changes in lung function and radiograph, and (3) to evaluate the relation of quartz exposure and retention to individual response leading to early silicosis. Thirty-six sheep were exposed by repeated intratracheal infusion at 10-day intervals to 100 mg Minusil-5 in 100 ml saline (Si group), and 10 sheep were exposed at the same intervals to 100 ml saline (control). All sheep were investigated at 3-month intervals by chest radiograph, lung function, and lung lavage. At month 9, chest radiograph score of parenchymal opacities was significantly increased at 2.8 +/- 0.6 versus 0.4 +/- 0.4 in the Si group (p less than .05), establishing early radiologic silicosis. Lung function was significantly altered with reduction in lung compliance, vital capacity, and diffusion capacity (p less than .05). Lung lavage cellularity revealed significant increase in total cells (X 2.5), macrophages (X3), and neutrophils (X3). Albumin in BAL remained at the control level. Fibronectin production was significantly increased, as was the fibroblast growth activity, without significant change in procollagen 3 at this early stage of disease. Total phospholipids were significantly elevated in the Si-exposed sheep, and the profile demonstrated an increase in all the phospholipid components. Spontaneous release of hydrogen peroxide by alveolar cells was not increased, but in the presence of phorbol myristate acetate (PMA) higher levels of peroxide were found in the quartz-exposed sheep (p less than .05). The cellular and biochemical alterations of lung lavage preceded other changes. At month 12, there were good correlations (r greater than .49, p less than .001) between parameters evaluating related phenomena but poor correlations between measurements evaluating different aspects of the disorder. To investigate the heterogeneity in the individual response of sheep to the same exposure (susceptibility), individual quartz retention levels at month 12 were measured and found to correlate well with individual parameters of disease activity. We concluded that in early silicosis of sheep, cellular and biochemical changes in lung lavage preceded derangements of pulmonary function and radiographic abnormalities. Thereafter, parameters of lung lavage, lung function, and radiograph were significantly interrelated, but for a given exposure the degree of quartz retention appeared to determine the intensity of the silicotic process. PMID- 2545437 TI - Effect of reduced phytate wheat bran on zinc absorption. AB - The rise in plasma zinc after the ingestion of 765 mumol (50 mg) elemental zinc was measured over 6 h in three groups of healthy subjects who, in a second test, also consumed a standard wheat bran, a reduced phytate, high fibre processed bran or rice-based, low phytate, low fibre breakfast cereal (Rice Krispies). Standard bran, reduced phytate bran and Rice Krispies all significantly decreased the area under the plasma zinc time curve (AUC) compared to the zinc alone test. The AUC obtained with standard bran was significantly lower than that after reduced phytate bran (mean +/- s.e.: -5.9 +/- 2.7 vs 18.5 +/- 4.2 mumol.h/l, P less than 0.001). The percentage reductions in AUC produced by the test meals were: standard bran 106.4 +/- 2.4, reduced phytate bran 75.9 +/- 4.8, Rice Krispies 46.3 +/- 10.4 per cent, P less than 0.01). Histidine, taken with standard bran in a small subgroup of subjects, tended to improve zinc absorption but the difference was not statistically significant. PMID- 2545438 TI - Specificity of binding of clathrin adaptors to signals on the mannose-6 phosphate/insulin-like growth factor II receptor. AB - Adaptors mediate the interaction of clathrin with select groups of receptors. Two distinct types of adaptors, the HA-II adaptors (found in plasma membrane coated pits) and the HA-I adaptors (localized to Golgi coated pits) bind to the cytoplasmic portion of the 270 kd mannose 6-phosphate (M6P) receptor-a receptor which is concentrated in coated pits on both the plasma membrane and in the trans Golgi network. Neither type of adaptor appears to compete with the other for binding, suggesting that each type recognizes a distinct site on the M6P receptor tail. Mutation of the two tyrosines in the tail essentially eliminates the interaction with the HA-II plasma membrane adaptor, which recognizes a 'tyrosine' signal on other endocytosed receptors (for example, the LDL receptor and the poly Ig receptor). In contrast, the wild type and the mutant M6P receptor tail (lacking tyrosines) are equally effective at binding HA-I adaptors. This suggests that there is an HA-I recognition signal in another region of the M6P receptor tail, C-terminal to the tyrosine residues, which remains intact in the mutant. This signal is presumably responsible for the concentration of the M6P receptor, with bound lysosomal enzymes, into coated pits which bud from the trans-Golgi network, thus mediating efficient transfer of these enzymes to lysosomes. PMID- 2545439 TI - PDGF A chain homodimers drive proliferation of bipotential (O-2A) glial progenitor cells in the developing rat optic nerve. AB - The bipotential glial progenitor cells (O-2A progenitors), which during development of the rat optic nerve give rise to oligodendrocytes and type 2 astrocytes, are stimulated to divide in culture by platelet-derived growth factor (PDGF), and there is evidence that PDGF is important for development of the O-2A cell lineage in vivo. We have visualized PDGF mRNA in the rat optic nerve by in situ hybridization, and its spatial distribution is compatible with the idea that type 1 astrocytes are the major source of PDGF in the nerve. We can detect mRNA encoding the A chain, but not the B chain of PDGF in the brain and optic nerve, suggesting that the major form of PDGF in the central nervous system is a homodimer of A chains (PDGF-AA). PDGF-AA is a more potent mitogen for O-2A progenitor cells than is PDGF-BB, while the reverse is true for human or rat fibroblasts. Fibroblasts display two types of PDGF receptors, type A receptors which bind to all three dimeric isoforms of PDGF, and type B receptors which bind PDGF-BB and PDGF-AB, but have low affinity for PDGF-AA. Our results suggest that O-2A progenitor cells possess predominantly type A receptors, and proliferate during development in response to PDGF-AA secreted by type 1 astrocytes. PMID- 2545440 TI - Cell type specific trans-acting factors are involved in alternative splicing of human fibronectin pre-mRNA. AB - ED-A and ED-B are facultative type III homologies of fibronectin, encoded by alternatively spliced exons, described in man and in rat. A hybrid alpha-globin fibronectin minigene containing the ED-B region from the human gene has been transfected in human cell lines derived from various tissues, in order to study the processing of the generated precursor RNA in the different cell environments. In most tested lines the pre-RNA is alternatively spliced and produces two mature RNAs, with and without the ED-B exon, in different ratios that closely resemble the corresponding endogenous fibronectin RNAs. In a hepatoma cell line, Hep 3B, only one RNA is produced, in which the ED-B exon is absent; the same pattern of splicing is observed in liver. The data show that all the information required to produce accurate and regulated alternative splicing of the ED-B exon is contained in the fragment used and cell specific factors are necessary for the pre-RNA to be differentially spliced in the various cell lines. In contrast, expression in Hep 3B of a similar gene containing the ED-A area failed to reproduce the liver specific splicing pattern. Therefore regulation of ED-A processing is likely to involve different mechanisms to those responsible for control of ED-B splicing. PMID- 2545441 TI - A C-terminal domain of GAP is sufficient to stimulate ras p21 GTPase activity. AB - The cDNA for bovine ras p21 GTPase activating protein (GAP) has been cloned and the 1044 amino acid polypeptide encoded by the clone has been shown to bind the GTP complexes of both normal and oncogenic Harvey (Ha) ras p21. To identify the regions of GAP critical for the catalytic stimulation of ras p21 GTPase activity, a series of truncated forms of GAP protein were expressed in Escherichia coli. The C-terminal 343 amino acids of GAP (residues 702-1044) were observed to bind Ha ras p21-GTP and stimulate Ha ras p21 GTPase activity with the same efficiency (kcat/KM congruent to 1 x 10(6) M-1 s-1 at 24 degrees C) as GAP purified from bovine brain or full-length GAP expressed in E. coli. Deletion of the final 61 amino acid residues of GAP (residues 986-1044) rendered the protein insoluble upon expression in E. coli. These results define a distinct catalytic domain at the C terminus of GAP. In addition, GAP contains amino acid similarity with the B and C box domains conserved among phospholipase C-II, the crk oncogene product, and the non-receptor tyrosine kinase oncogene products. This homologous region is located in the N-terminal half of GAP outside of the catalytic domain that stimulates ras p21 GTPase activity and may constitute a distinct structural or functional domain within the GAP protein. PMID- 2545442 TI - The quaternary structure of Tet repressors bound to the Tn10-encoded tet gene control region determined by neutron solution scattering. AB - The spatial arrangement of Tet repressor dimer, both free and in complex with an 80 bp DNA fragment spanning the wild-type Tn10-encoded tet transcriptional control sequence containing a tandem repeat of two operators, has been determined by neutron small-angle scattering. The active, free Tet repressor dimer is an elongated and flat molecule with a maximum dimension of 11 +/- 1.5 mm which can be approximated by an ellipsoid with the half-axes 6 nm, 2.5 nm and 1 nm. The overall conformation undergoes no detectable change when the repressor dimer is bound to a DNA fragment containing a single tet operator. The normal distance between the centre of gravity of the protein and the DNA axis is 3.0 +/- 0.1 nm, indicating that the repressor dimer is mainly located on one side of the DNA. When bound to the wild type tet control DNA, the two repressor dimers have a centre-to-centre distance of 11.0 +/- 0.5 nm. Their minimal distance is 5 +/- 2 nm. Protein-protein contacts via loop formation of the DNA by repressor binding is excluded. The repressors are well separated and have no direct contact. A model is proposed where the two repressor dimers are located on opposite sides of the DNA and the DNA is not strongly bent in the complex. PMID- 2545444 TI - Medical therapy of VIPomas. AB - VIP-secreting tumors are rare, but they produce a dramatic clinical picture, the most prominent feature of which is profuse, watery diarrhea and hypokalemia. A definitive diagnosis is aided by the determination of plasma VIP concentrations through the use of the sensitive radioimmunoassays that are now available. Intestinal secretion resulting from the direct action of VIP on the intestinal epithelial cell receptors accounts for the loss of fluid and electrolytes in patients with VIPoma. The hypokalemia is the result of passive and VIP-induced active secretion of potassium by colonic epithelial cells. Surgery is the most definitive treatment of VIPoma; pharmacotherapy is extremely important in controlling symptoms and stabilizing the patient prior to surgery. Sandostatin and glucocorticoids are well established agents in the management of the secretory diarrhea; other pharmacologic agents, including clonidine, indomethacin, phenothiazines, lithium carbonate, and propranolol, may be helpful in selected patients but require further study. The most potent and promising drug for the treatment of VIPoma is a new peptidomimetic agent--Sandostatin. This metabolically stable synthetic analogue of somatostatin appears, in part, to inhibit the release of VIP from the tumor and the secretion of chloride by the intestine. In addition to controlling the diarrhea, it may have a direct effect on the tumor in reducing its size. PMID- 2545443 TI - Activity of the transposon Tam3 in Antirrhinum and tobacco: possible role of DNA methylation. AB - The transposon Tam3 from Antirrhinum majus can transpose in a heterologous host (Nicotiana tabacum); thus the element is autonomous, probably encoding the specific information required for its own transposition. In transgenic tobacco Tam3 rapidly becomes methylated at its ends whilst adjacent flanking sequences remain hypomethylated. This methylation may account for our failure to detect Tam3 transposition in the progeny of transgenic tobacco. Treatment with the inhibitor of cytosine methylation, 5 aza-cytosine appeared to induce transposon related activity at a low level. In Antirrhinum methylation also appears to be associated with inactivation of Tam3 copies. PMID- 2545445 TI - Mammographic wire-guided biopsies in non-palpable breast lesions. AB - The use of mammography, both for diagnostic and screening purposes, is steadily increasing. It is a very sensitive examination method; findings such as an ill defined or spiculated mass, clustered, irregular and dense microcalcifications, asymmetric densities or architectural distortion, are suggestive of malignancy. These findings are often not specific enough to make an unequivocal diagnosis of malignancy, and they are frequently not palpable. The presence of clinically occult, but mammographically present findings warrants biopsy to determine their histological nature. In order to avoid unnecessarily large biopsies and to assure the removal of the suspicious portion of the breast, precise localization in necessary, guided by mammography. Using a needle-hookwire method, 87 biopsies were done for non-palpable breast lesions detected among 3,196 mammographies performed. Experience with this method and the results show that 17 patients, i.e. every fifth biopsy, had a malignancy. In 5 patients out of 17, the findings proved to be in situ carcinoma. In another group of 17 women the biopsy result was atypical epithelial proliferation, considered to be a transitional phase of in situ carcinoma. PMID- 2545446 TI - Type I and I-trimer collagens as substrates for breast carcinoma cells in culture. Effect on growth rate, morphological appearance and actin organization. AB - Type I-trimer collagen, isolated from biopsy fragments of ductal infiltrating carcinomas, was used as a substrate for human breast carcinoma cells in long-term culture to monitor growth rate, morphological appearance and actin organization in comparison with normal type I collagen and plain plastic. After 11 days of culture, type I-trimer collagen exerts a more pronounced effect on cell proliferation, leading to a final increment of cell population of 35% versus regular type I substrate. Furthermore, type I-trimer collagen induces cell motility, as testified by morphological appearance and actin immunofluorescence test. On the basis of the in vitro results, it is postulated that in vivo the stromal areas containing trimer collagen, rather than repressing invasive growth, may provide a more suitable environment for tumor proliferation and spreading-out with respect to regular type I. PMID- 2545447 TI - Mechanism of action of Clostridium difficile toxin B: role of external medium and cytoskeletal organization in intoxicated cells. AB - Toxin B, an exotoxin produced by Clostridium difficile, induces the rounding-up and arborization of cultured mammalian cells, a typical effect which resembles that provoked by cytochalasins. In this study, the effect of toxin B was examined on astroglial cells grown in primary culture. A specific antiserum to toxin B was used to investigate its mechanisms of action. We found that the toxin exerts its effects on cell morphology after its incorporation into cells. The internalization of toxin B requires the presence of calcium ions in the extracellular medium. Replacement of NaCl with sucrose or with potassium glutamate prevents the internalization of the toxin. The direct introduction of calcium ions into cells by the calcium ionophore A23187 stimulates toxin-induced morphological changes. In contrast, toxin-induced morphological transformations were prevented in cells treated with tumor-promoting phorbol. esters or with dibutyryl-cAMP, although such treatment did not abolish the internalization of the toxin. As in the other cell types, the earliest effect of toxin B on astrocyte cytoskeleton is the disruption of actin filaments, without no visible alteration of intermediate filament nor microtubule networks. As astrocytes with toxin-induced stellate morphology survive toxin treatment, the progression of cell morphology and cytoskeleton organization were followed for several weeks. Twenty-six days after exposure to toxin B, stellate astrocytes have processes which were markedly longer and much more branched than those of cells freshly exposed to toxin. At that time, cells are still devoid of F-actin as assessed with rhodamine-conjugated phalloidin and only 70% contain vimentin while all astrocytes present in control cultures express vimentin. Some flat epithelioid astrocytes with prominent bundles of microfilaments reappear during the second week after toxin treatment. Our results show that Clostridium difficile toxin B is internalized into brain astrocytes in culture where it acts by modifying cytoskeletal elements. Its cytopathic effects are reversible. Although actin related components of the cytoskeleton are the major target of toxin B, other cytoskeletal elements also seem to be affected. PMID- 2545448 TI - Involvement of cytoplasmic membranes in the non-lytic infection of K-562 cells by Semliki Forest virus. AB - An analysis of the human leukemia cell line, K-562, infected with Semliki Forest virus, has been made with transmission electron microscopy. In contrast to the usual surface budding of the enveloped virus on the plasma membrane of vertebrate cells leading to cytolysis within 20 h, K-562 cells do not show surface budding, and the cells remain intact for periods of several months. Several unusual features of the infection include: 1) the rough endoplasmic reticulum arranges early into continuous perinuclear chains; 2) during the time of virus replication and release, the nucleocapsids aggregate on the cytoplasmic side of internal vesicles in the region of the cell where the Golgi complex is normally located; and 3) during this same time period, the vesicles are seen to contain enveloped virions and rod-like formations, a result suggesting that budding has occurred into these vesicles. Viruses are presumably released from the cell as these vesicles fuse with the plasma membrane. By 12 days post-infection and thereafter, the intact cells show electron-dense aggregates of chromatin, large vacuoles and lipid inclusions throughout the cytoplasm, and only a few virion-containing vesicles. PMID- 2545449 TI - Recurrent ascites in an infant with perinatally acquired cytomegalovirus infection. AB - We describe an infant with symptomatic perinatally-acquired cytomegalovirus (CMV) infection manifested by fever, anaemia, thrombocytopenia and hepatosplenomegaly. This infant developed recurrent episodes of severe ascites during which the virus was isolated from his urine. This rare hepatic manifestation of neonatal CMV infection has, to the best of our knowledge, only been reported twice in aborted fetuses with intrauterine systemic CMV infection. PMID- 2545450 TI - Structural alteration of the insulin-like growth factor II-gene in Wilms tumour. AB - In this study messenger ribonucleic acid (mRNA) and DNA of five Wilms tumours were investigated. As expected, the level of insulin-like growth factor (IGF) II mRNA was elevated up to 50 times in tumour tissue as compared to normal adjacent kidney tissue. In addition, genomic DNA was isolated and digested with appropriate restriction enzymes. Southern blots were prepared and hybridized to IGF II-cDNA probes. Additional bands were present in one of the five Wilms tumours compared to normal tissue. The results indicate a rearrangement of the IGF II-gene on one of the two chromosomes. It is speculated, that this change is responsible for the elevated IGF II expression which may be a factor contributing to tumour growth. PMID- 2545452 TI - Coxsackie B3 virus encephalitis in a patient with agammaglobulinaemia. AB - A 19-month-old boy with agammaglobulinaemia contracted a Coxsackie B3 virus infection which caused meningoencephalitis. His neurological status deteriorated over the following 4 years and he died aged 6 years. Treatment with gammaglobulin i.m. and IgG-Fab2 fragments i.v. was not effective. Enterovirus can cause life threatening infections in immunodeficient patients. PMID- 2545451 TI - A trial of RIT-4237 rotavirus vaccine in 1-month-old infants. AB - In a double blind placebo-controlled study rotavirus vaccine RIT 4237 was offered to young infants after the fourth week of life. The vaccine was very well tolerated. Fifty-seven out of 100 vaccine recipients and 10 out of 103 placebo recipients developed rotavirus-IgM-antibodies during the 1 month follow-up period. During a 6 month follow-up, stool samples from 2 out of 12 vaccinees and from 5 out of 12 placebo recipients contained rotavirus. PMID- 2545453 TI - Involvement of the caval vein lumen by a metastasis of a non-seminomatous testicular tumor. AB - We present here a case of metastatic non-seminomatous testicular tumor invasion of the lumen of the inferior caval vein. Treatment was surgical removal with lymph-node debulking, completed by BEP chemotherapy. PMID- 2545454 TI - Enalapril and hydrochlorothiazide in hypertensive Africans. AB - The antihypertensive efficacy both of angiotensin converting enzyme (ACE) inhibitors and thiazide diuretics has been claimed to be influenced by plasma renin activity, which declines with age and is low in blacks. In a double-blind, placebo-controlled, double-dummy, randomized, parallel-group preliminary study, the antihypertensive efficacy and tolerability of the ACE inhibitor enalapril (20 mg day-1) and hydrochlorothiazide (50 mg day-1) were evaluated and compared for 4 weeks in 20 African patients with essential hypertension. The two groups had similar baseline clinical features and serum Na+ and K+ levels. Hydrochlorothiazide caused a significant and sustained fall in erect blood pressure with a reflex tachycardia. Enalapril exerted only a modest antihypertensive action, but significantly reduced erect heart rate. Direct comparison of hydrochlorothiazide- and enalapril-induced hypotension suggested a greater fall in subjects on the thiazide. The 95% confidence limits for the thiazide-enalapril difference in antihypertensive action at the end of the study was 39.5 to -7.5 mm Hg systolic and 22.0 to -6.6 mm Hg diastolic. The maximal blood pressure fall after hydrochlorothiazide was positively correlated with age (r = 0.50; p less than 0.05), whilst that of enalapril was inversely related age to (r = -0.57, p less than 0.05). The results are compatible with the notion that ACE inhibitor monotherapy may be less effective than thiazide diuretic treatment in African and black patients with essential hypertension. The findings also support the concept that age and racial factors may influence the response to antihypertensive treatment. PMID- 2545455 TI - Beta-adrenoceptor responses to inhaled salbutamol in normal subjects. AB - The aim of the present study was to quantify and compare the airways and systemic beta-adrenoceptor responses to inhaled salbutamol in normal subjects. Seven non atopic, normal subjects were given cumulative doubling doses of inhaled salbutamol (100 micrograms to 4000 micrograms) or placebo in a single-blind cross over design. Airways (sGaw, FEF 50%, FEF 25%), tremor, haemodynamic and metabolic responses were measured at each dose increment. There were dose-related changes in sGaw, FEF 50% and FEF 25% up to a plateau at 1.0 mg. Analysis of individual responses showed that most subjects required either 1.0 or 2.0 mg for maximum bronchodilatation, independent of the parameter of airflow. There was no correlation between maximum bronchodilatation, independent of the parameter of airflow. There was no correlation between maximum response and baseline airway calibre. In contrast to airways effects, systemic beta-adrenoceptor responses did not occur until 500 micrograms, and a ceiling in the dose-response curve was not reached. There were significant correlations between airways, tremor and haemodynamic responses, and between different metabolic variables. The intraindividual variability was greatest for tremor and sGaw, although this was small in comparison to the size of maximum change with salbutamol. The converse applied to the hypomagnesaemic response. PMID- 2545456 TI - Thromboxane A2 activates phospholipase C in astrocytoma cells via pertussis toxin insensitive G-protein. AB - The properties of thromboxane A2 (TXA2) receptors were examined in 1321N1 human astrocytoma cells. 9,11-Epithio-11,12-methanothromboxane A2 (STA2), a stable analogue of TXA2, stimulated the accumulation of inositol phosphates (IPs) with an EC50 of about 50 nM. The STA2-induced accumulation of IPs was inhibited concentration dependently by ONO3708, a TXA2 receptor antagonist, with an inhibition constant (Ki) of about 10 nM. Inositol trisphosphate (IP3) was accumulated more rapidly than inositol bisphosphate (IP2) in response to STA2. HPLC analysis indicated that inositol 1,4,5-trisphosphate accumulated in the presence of STA2. STA2 alone had no effect on the accumulation of IPs in membrane preparations but it potentiated the accumulation induced by GTP gamma S. [3H]SQ29548, a TXA2 receptor antagonist, bound specifically to TXA2 receptors, expressing a single binding site with a dissociation constant (Kd) of 10.9 nM. The competition curve for STA2 inhibition of [3H]SQ29548 binding was shifted to the right and was steeper in the presence of GTP gamma S. Pertussis toxin (IAP) elicited ADP-ribosylation of 41KD protein but had no effect on the sensitivity to GTP of the STA2 inhibition of SQ29548 binding or of STA2-induced accumulation of IPs. It is concluded from these results that the stimulation of TXA2 receptors results in activation of phospholipase C via a GTP binding protein and that the protein is not a substrate for IAP. PMID- 2545457 TI - Receptors for kinins in isolated arterial vessels of dogs. AB - A variety of kinin peptides, agonists and antagonists were tested with dog carotid and renal arteries in order to characterize kinin receptor types and functions. The dog carotid artery responds to bradykinin with concentration dependent relaxation only when the endothelium is intact but des-Arg9-bradykinin is practically inactive. The effect of bradykinin is blocked by B2 receptor antagonists, suggesting that the dog carotid artery has B2 receptors in the endothelium. These receptors mediate relaxation of the arterial smooth muscles by promoting the release of an endothelium-derived relaxing factor whose action is prevented by methylene blue. Kinins relax the dog renal artery with or without endothelium. Methylene blue prevents the effect of bradykinin only, suggesting that B2 receptors, promoting the release of endothelium-derived relaxing factor, are present in the endothelium of the dog renal artery. Moreover, the dog renal artery appears to have both B2 and B1 receptors mediating relaxation of the arterial smooth muscle. The presence of the two receptor types has been demonstrated by means of specific agonists and antagonists. Indomethacin blocks the effects of both bradykinin and des-Arg9-bradykinin on the dog renal artery without endothelium, suggesting that muscular B1 and B2 receptors act by promoting the release of prostaglandins. Captopril, a kininase II inhibitor, potentiates the effect of bradykinin on the dog carotid artery more than on the dog renal artery. PMID- 2545458 TI - Antagonist profile of 6,7-dichloro-3-hydroxy-2-quinoxalinecarboxylate at excitatory amino acid receptors in the neonatal rat spinal cord. AB - In the neonatal rat spinal cord, 6,7-dichloro-3-hydroxy-2- quinoxalinecarboxylate antagonised responses mediated at both N-methyl-D-aspartate (NMDA) and non-NMDA receptors. The antagonism of responses to NMDA was unsurmountable and mediated via an antagonist action at the allosterically-linked strychnine-insensitive glycine site. At non-NMDA receptors, 6,7-dichloro-3-hydroxy-2 quinoxalinecarboxylate appeared to act as a competitive antagonist at low concentrations and a non-competitive antagonist at higher concentrations. In contrast to published data, this antagonist did not distinguish between the responses mediated by DL-alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate (AMPA) and kainate. PMID- 2545459 TI - Sumatriptan (GR 43175) interacts selectively with 5-HT1B and 5-HT1D binding sites. AB - The ability of sumatriptan (GR 43175; 3-[2-dimethylamino]ethyl-N-methyl-1H-indole 5 methane sulphonamide) to interact with 13 neurotransmitter receptor sites was determined using radioligand binding techniques. Sumatriptan displayed the highest affinity for 5-HT1D (Ki = 17 nM) and 5-HT1B (Ki = 27 nM) binding sites and was slightly less potent at 5-HT1A binding sites (Ki = 100 nM). By contrast, sumatriptan was essentially inactive (Ki greater than 10,000 nM) at each of the 10 other binding sites analyzed. These data indicate that sumatriptan interacts selectively with 5-HT1B and 5-HT1D sites and suggest that these interactions may be the basis of its apparent efficacy in the acute treatment of migraine. PMID- 2545461 TI - Inhibition of cholinergic and non-adrenergic, non-cholinergic bronchoconstriction in the guinea pig mediated by neuropeptide Y and alpha 2-adrenoceptors and opiate receptors. AB - The mechanisms underlying the regulatory influence of neuropeptide Y (NPY) and of alpha 2-adrenoceptor and opiate receptor activation on cholinergic and excitatory non-adrenergic, non-cholinergic (e-NANC) neurotransmission were studied in guinea pig hilus bronchi in vitro. NPY inhibited both the cholinergic and e-NANC bronchial contractions evoked by field stimulation. The NPY attenuation of the e NANC contraction could not be antagonized by the alpha 2-antagonist, idazoxan, or naloxone. UK 14,304 a specific alpha 2-agonist, also reduced the two nervous components of bronchial contraction and this action was inhibited by idazoxan. NPY and UK 14,304 exerted a minor influence on the bronchial smooth muscle tone per se or on contractions evoked by acetylcholine or neurokinin A. This suggested that the inhibitory responses were caused by a prejunctional action reducing the release of transmitter substances from sensory and cholinergic nerve endings. Furthermore NPY (10(-7) M) seemed to be more potent to inhibit both contractile components than noradrenaline (10(-6) M) in the presence of propranolol (3 X 10( 6) M). Morphine was able to reduce the e-NANC response via a naloxone-sensitive mechanism. The capsaicin-evoked bronchoconstriction and the bronchodilator NANC effect evoked by field stimulation were, however, not influenced by UK 14,304. It is concluded that NPY, alpha 2-receptor and opiate receptor activation inhibit the release of sensory transmitters evoked by field stimulation but not by capsaicin. PMID- 2545460 TI - Rapid desensitization and down-regulation of 5-HT2 receptors by DOM treatment. AB - The regulation of the 5-HT2 receptor-mediated head twitch response and of 5-HT2 receptor binding in the frontal cortex was studied in rats treated repeatedly with the 5-HT2 agonist 1-(2,5-dimethoxy-4-methylphenyl)-2-aminopropane (DOM) (2.5 mg/kg, s.c.). Four injections in 24 h produced a near maximal reduction in the behaviour (-70%) and in the Bmax for [3H]ketanserin binding (-41%). The KD values tended to increase slightly. 5-HT2 receptors reappeared, with half-lives of 5.5 to 3 days. In view of the reported anomalous 5-HT2 receptor regulation by antagonists and the regular regulation by agonists, we propose a refinement in the receptor regulation theory. PMID- 2545462 TI - Beta-adrenoceptor-effector system of the human macrophage U937 cell line. AB - Like animal peritoneal macrophages, cells of the U937 human macrophage cell line were found to possess beta-adrenoceptors with a density of approximately 1400 sites/cell of the beta 2-subtype. Agonist occupancy increased adenylate cyclase activity and lead to rapid sequestration of cell surface receptors, functional desensitization, and receptor downregulation. Thus human macrophages possess beta adrenoceptors and the U937 cell provides a system for studying the interactions of this receptor with macrophage immune function. PMID- 2545464 TI - Presence of specific platelet-activating factor binding sites in the rat retina. AB - The specificity of the effects of platelet-activating factor (PAF) on the electrophysiology of the retina suggests the existence of specific sites for PAF in this tissue. In this study, we report the presence of tritiated PAF ([3H]PAF) specific binding sites in membrane preparations of the retina of albino rats. The binding of [3H]PAF was saturable, specific, time-dependent and reversible. Scatchard analysis of the data revealed that the high-affinity retinal binding site possessed a Kd of 2.9 +/- 0.4 nM and Bmax of 0.85 +/- 0.16 pmol/mg protein. These values are comparable with those found for the membranous PAF receptor sites in platelets, neutrophils, lung tissue and brain. We have recently reported that PAF dose dependently modulates the b-wave of the electroretinogram (ERG) obtained from the isolated rat retina. The results of the present study suggest that such PAF-induced disturbances of the ERG may be mediated via specific receptors located in the retina. PMID- 2545463 TI - Pharmacologic profile of lipoxins A5 and B5: new biologically active eicosanoids. AB - In this study, we have investigated the biological activities of LxA5 and LxB5 in isolated smooth muscle preparations of the guinea pig. LxA5 slowly contracted pulmonary parenchymal strips isolated from guinea pigs in a concentration dependent manner over the range of 0.1-2.2 microM. This bronchoconstrictor effect was not associated with release of peptide leukotrienes (i.e. LTC4, LTD4 and LTE4) and only a slight increase in thromboxane B2. Furthermore, the bronchoconstrictor effect was not blocked by lipoxygenase inhibitors, suggesting the pulmonary effect is not mediated by lipoxygenase products. However, a peptide leukotriene receptor antagonist (e.g. SK&F-104353) inhibited or reversed the LxA5 response indicating that LxA5 and the peptide leukotrienes may share the same receptor. In contrast to LxA5, LxB5 displayed no significant bronchoconstrictor effect at concentrations up to 2.2 microM. Moreover, LxA5 and LxB5 did not exert a significant endothelium-dependent vasorelaxation in aortic vascular smooth muscle as do LxA4 and LxB4. Thus, LxA5 and LxB5 display a unique biological profile which differs from LxA4 and LxB4. LxA4 may be a mediator in circulatory disease states (e.g. myocardial ischemia, circulatory shock), but LxA5 and LxB5 are not as likely candidates as mediators of disease. PMID- 2545465 TI - Species relationships for plasma angiotensin converting enzyme activity using a furanacryloyl tripeptide substrate. AB - Angiotensin converting enzyme (ACE; EC 3.4.15.1) activities were compared in plasma samples obtained from three species using a furanacryloyl tripeptide substrate. The enzyme activity observed in Wistar rat plasma was higher than the activities observed in the other two species. Using this substrate, human and canine plasma enzyme activities were similar-unlike published data where hippuryl histidyl-leucine was used as substrate. PMID- 2545466 TI - Pathology of spontaneous malignant fibrous histiocytoma in a Japanese white rabbit. AB - Spontaneous malignant fibrous histiocytoma (MFH) of the chest wall was found in a 10-year-old male Japanese white rabbit. Histologically, the MFH consisted mainly of areas of storiform, pleomorphic and myxoid patterns. Positive reactions for acid phosphatase (Ac-P), non-specific esterase (N-SE) and beta-glucuronidase (beta-GL) were demonstrated in fibroblast-like, histiocyte-like and giant cells. Moreover, a strongly positive fibronectin (FN) reaction was observed mainly in histiocyte-like and giant cells. In electron microscopy, tumor cells were composed morphologically of various types of cells such as fibroblast-like, histiocyte-like, undifferentiated and giant cells. This case was quite similar to those reported in man or other animals. PMID- 2545467 TI - NMDA receptor antagonists can enhance or impair learning performance in animals. AB - The effects of NMDA receptor antagonism on learning and memory were investigated using competitive (DL-2-amino-7-phosphonoheptanoate, AP7) and non-competitive (MK 801) blockers in three different learning tasks. Administration (i.p.) of drugs prior to training resulted in impaired learning performance in the place navigation and dark-avoidance paradigms, and improved performance in the step down passive avoidance task; however, using this treatment protocol, the possibility of drug-induced non-mnemonic effects modifying learning performance could not be excluded. Drug administration immediately post-trial had no effect in the place-navigation paradigm, and improved retention performance in the dark avoidance and step-down avoidance tasks. The similar results obtained with both types of antagonist indicate that the observed effects are indeed due to NMDA receptor blockade, and hence that such blockade modifies learning in a task dependent manner. Exclusion of non-mnemonic effects by using the post-trial treatment regime demonstrates that NMDA antagonists facilitate learning of passive avoidance tasks. PMID- 2545469 TI - [Does ethanol potentiate GABAergic inhibition?]. AB - The effect of systemically and microelectro-osmotically applied ethanol on sensitivity of sensorimotor cortical neurons of unanesthetized rabbit brain to microiontophoretically administered GABA and glycine was studied. No correlation was found between responses of single neurons to ethanol, GABA and glycine. The inhibitory responses to ethanol were summed with responses to GABA and glycine (additive interaction). Ethanol failed to potentiate GABA effect. PMID- 2545470 TI - [The effects of nephrectomy and angiotensin converting enzyme inhibitor on production and clearance rates of angiotensinogen in dogs]. AB - The clearance and production rates of plasma angiotensinogen of normal dog, nephrectomized dog and angiotensin converting enzyme inhibitor (ACEI) administered dog were calculated. Plasma angiotensinogen was measured by adding excessive dog renin to the plasma sample. The clearance rate was 4.4, 4.0 and 3.8 ml/kg/hr and the production rate was 9.6, 21.6 and 7.6 micrograms/kg/hr, respectively. After nephrectomy, the clearance rate of angiotensinogen decreased only 10 percent but the production rate increased more than twice. These results mean that the kidney does not play an important role in the clearance of angiotensinogen but has some influence on its production. This increase in production rate due to nephrectomy may mean that the kidney has some inhibitory effect on the secretion of angiotensinogen from the liver but this phenomenon is not able to explain by much less hydrolysis of angiotensinogen. By the administration of ACEI, not only the clearance rate but also the production rate of plasma angiotensinogen decreased as compared with control dog, but the balance between them resulted in the decreased plasma angiotensinogen concentration. These results show that the decreased production rate was greater than the decreased clearance rate in ACEI administered dog. PMID- 2545468 TI - GABA[A] receptor level changes in female hamster forebrain following in vivo estrogen progesterone and benzodiazepine treatment: a quantitative autoradiography analysis. AB - The levels of gamma amino butyric acid (GABAA) receptors (i.e. 3H-Muscimol binding sites) were determined by quantitative neurotransmitter receptors autoradiography in ovariectomized (OVX), OVX-estradiolo (E), OVX-progesterone (P), OVX-E + P, diazepam (DZ) and DZ + Ro15-1788 treated female hamsters. The various hormonal treatments altered 3H muscimol binding in many brain areas, whereas DZ and DZ + Ro15-1788 had little influence on GABAA receptor levels at the onset of the blocked aggressive behavioral activity. For example, E, P and E + P all significantly increased 3H muscimol binding in medial preoptic area, ventromedial hypothalamic nuclei, and vertical diagonal bandmedial septal nucleus, whereas P treatment increased binding in the caudate-putamen and decreased it in reuniens nucleus of the thalamus. E and E + P treatments increased 3H muscimol binding in the corticomedial amygdala nucleus and hippocampus. Diazepam treatment decreased the GABAA receptor binding in the caudate-putamen and basolateral amygdala, while having no effect in the other brain regions where hormone treatment was effective. In vitro incubation of brain sections with micromolar concentrations of E or P did not change muscimol binding. These results suggest that ovarian steroids, and benzodiazepines to a lesser extent, modulate 3H muscimol binding but do so in different brain regions. The hormone effects on 3H muscimol binding in the critical reproductive centres at a time period that coincides with the onset of its behavioral effect is consistent with a genomic controlled activity. PMID- 2545471 TI - [Dermal fibrous histiocytoma with inductive formation of a partially pigmented basalioma of the trichoepitheliomatous type]. AB - Report about a 73 years old woman with a rare dermal tumor of the right shoulder. Histological examination of the tumor shows a pigmentary basal cell carcinoma on type of trichoepithelioma overlying long-standing dermal fibrous histiocytoma. PMID- 2545472 TI - Effects of lectins with various carbohydrate binding specificities on lipid metabolism in isolated rat and hamster adipocytes. AB - 1. Mannose-binding and N-acetylglucosamine binding lectins exhibited potent antilipolytic and lipogenic activities. 2. Fucose-binding lectins had minimal lipogenic activity but possessed antilipolytic activity. 3. Most galactose binding and N-acetylgalactosamine-binding lectins were devoid of significant antilipolytic and lipogenic activities. Notable exceptions were lectins from Momordica charantia, Wisteria floribunda, Vicia villosa, Codium fragile and the Siberian pine tree. lipogenic activity but lacked antilipolytic activity. 5. The galactose-binding horse gram and osage orange lectins exerted antilipolytic activity in hamster but not in rat adipocytes. PMID- 2545474 TI - Temporal changes in phosphoamino acid phosphatase activities in murine erythroleukaemic cells. AB - 1. The activities of phosphotyrosine, phosphothreonine and phosphoserine phosphatases were measured at various time periods in Friend murine erythroleukaemic (MEL) cells. 2. Effects of DMSO (dimethyl sulphoxide) and HMBA (hexamethylene bisacetamide), inducers of differentiation, were examined. 3. The activities of all three enzymes showed cyclic variation when measured on a daily basis over a period of several days; this was also the case when phosphothreonine phosphatase was determined in cells treated with DMSO or HMBA and when phosphoserine phosphatase was assayed after stimulation of the cells with HMBA. 4. Evidence for rhythmic variation in phosphotyrosine phosphatase activities was also obtained when the activities were determined at hourly intervals both in control cells and those treated with HMBA. 5. The time-dependent changes observed could be significant in that control of dephosphorylation may possibly be achieved by altering the rhythms. PMID- 2545473 TI - Stimulation of inositol phospholipid breakdown in pig brain miniprisms by carbachol and monoamines: effect of K+. AB - 1. The effects of carbachol, monoamines and K+ upon the rate of inositol phospholipid breakdown in pig brain miniprisms have been investigated. 2. In the striatum, carbachol (EC50 approx. 1 microM) and noradrenaline (EC50 approx. 25 microM) stimulated inositol phospholipid breakdown, whereas 5-hydroxytryptamine (1-1000 microM) was without effect. 3. The rate of inositol phospholipid breakdown was increased by raising the assay [K+] to greater than or equal to 40 mM. In the hippocampus and hypothalamus, a synergistic effect between K+ and carbachol was noted, whereas in the striatum, the effects were additive. 4. In striatal and hippocampal miniprisms, dopamine also increased inositol phospholipid breakdown, albeit only at high (greater than or equal to 1 mM) concentrations. Dopamine (1 mM) reduced the stimulation produced by noradrenaline (1 mM), suggesting that the effect of dopamine is due to a weak noradrenergic action of this catecholamine. PMID- 2545475 TI - Regulation of the mammalian cytochrome P1-450 (CYP1A1) gene. PMID- 2545476 TI - Effects of metabolic acidosis and diabetes on the abundance of specific renal mRNAs. AB - 1. The effects of exogenously (NH4Cl ingestion) and endogenously (streptozotocin diabetes) generated chronic metabolic acidosis on the abundance of rat renal mRNAs have been examined. 2. Total RNA was translated in vitro and the translation products analyzed by two-dimensional gel electrophoresis. 3. The translation product identified as phosphoenolpyruvate carboxykinase (PEPCK) increased 3.5-fold in both acidosis and diabetes. 4. This increase was not observed in diabetic rats treated with NaHCO3. 5. The abundance of one other translation product increased in acidosis. 6. That of 10 others increased in diabetes, several of which were elevated regardless of acid-base status. 7. The abundance of one translation product decreased in acidosis and diabetes but not in NaHCo3 treated diabetic rats, indicating acid-base regulation of this product. 8. The results establish that the acidosis response is limited to a small number of renal mRNAs and confirm that renal PEPCK is primarily regulated by changes in acid-base status. 9. They also indicate that diabetes affects the abundance of specific renal mRNAs through mechanisms independent of acid-base status. PMID- 2545477 TI - Interaction of phosphatidylinositol-4-monophosphate with a low activity form of DNA polymerase alpha: a potential mechanism for enzyme activation. AB - 1. DNA polymerase alpha isolated from Norman murine myxosarcoma exhibited two isozyme forms, one with low specific activity and low DNA binding affinity (A1), and one with high specific activity and high DNA binding affinity (A2). 2. DNA polymerase alpha A1, but not A2, showed a significant increase in specific activity after treatment with phosphatidylinositol, ATP and phosphatidylinositol kinase, or with phosphatidylinositol-4-monophosphate. 3. Treatment of DNA polymerase alpha A1 with the phospholipase C hydrolysis product of phosphatidylinositol-4-monophosphate, inositol-1,4-bisphosphate, was sufficient to effect the transient increase in activity of polymerase A1 to a form not chromatographically distinguishable from isozyme form A2. PMID- 2545478 TI - Levamisole inhibition of alkaline phosphatase and 5'-nucleotidase of bovine milk fat globule membranes. AB - 1. The effect of levamisole (LMS) on alkaline phosphatase (EC 3.1.3.1) and 5' nucleotidase (EC 3.1.3.5) activities of bovine milk fat globule membranes (MFGM) was examined. 2. LMS inhibited MFGM alkaline phosphatase activity in a concentration-dependent manner with 50% inhibition produced by 49 +/- 23 microM LMS. 3. 5'-Nucleotidase was resistant to LMS inhibition with 30.9% inhibition produced by 10 mM LMS, the highest concentration tested. 4. LMS was an uncompetitive inhibitor of MFGM alkaline phosphatase with a Ki of 45 +/- 6 microM. 5. The extent of LMS inhibition of alkaline phosphatase was dependent on the substrate utilized in the assay. 6. The effect of LMS on bovine MFGM alkaline phosphatase was similar to LMS effects on other mammalian alkaline phosphatases of liver/kidney/bone/placental isoenzyme origin. PMID- 2545479 TI - Further studies on a chemoattractant derived from Aleurites Fordii Hemsl. seed. AB - 1. Activities stimulatory to PMN superoxide generation and chemotactic function were present in two separate gel chromatographic fractions of AFH-S and were designated AFH-S1 (500 kD) and AFH-S2 (38 kD). 2. Characterization studies of these activities revealed physiologic and pharmacologic characteristics dissimilar to those of other previously studied chemoattractants such as bacterial peptide FMLP. PMID- 2545480 TI - Thyrotropin but not epidermal growth factor down-regulates the isozyme I (PKa I) of cyclic AMP-dependent protein kinases in dog thyroid cells in primary cultures. AB - The activity of the two cAMP-dependent protein kinases (PKa I and PKa II) was evaluated in dog thyroid cells in primary cultures after a 6-day growth period induced by either thyrotropin (TSH) or epidermal growth factor (EGF). Although the total PKa activity was not affected in cells cultured in the presence of TSH or EGF, their actions on the PKa I and PKa II expressions were significantly different. The activity of PKa I was strongly inhibited by TSH (70-80%) while with EGF it was either stimulated or unaffected with respect to controls. The two mitogens did not have a significant effect on the activity of PKa II. Forskolin (Fk) mimicked the effect of TSH. The expression of the two regulatory subunits (R I and R II), evaluated by the covalent binding of 8-azido-cAMP, was similar to the expression of the corresponding catalytic activities, suggesting a coregulation of the catalytic and regulatory subunits from the same isozyme. After chronic stimulation by TSH, differentiated dog thyroid cells are almost completely deprived of PKa I. PMID- 2545481 TI - Glucocorticoid responsiveness of mouse mammary tumor virus (MMTV) promoters in a down-transcription hepatoma tissue culture (HTC) variant. AB - Complement-mediated cytolysis of the mouse mammary tumor virus (MMTV)-infected rat hepatoma (HTC) cell line, M1.54, resulted in recovery of a mutant derivative, designated CR5, in which the magnitude of both basal and dexamethasone-induced proviral MMTV RNA expression was selectively reduced. Variant CR5 cells were transfected with a plasmid containing the glucocorticoid-regulated MMTV promoter linked to the neomycin resistance gene (pLNL). Half-maximal resistance to G418 killing was glucocorticoid inducible in both pLNL-transfected CR5 and M1.54 cells and was dependent on glucocorticoid receptor occupancy. The down-transcription of MMTV provirus sequences cannot be conferred to transfected genes driven by the same viral promoter suggesting that CR5 cells are defective in cis acting factors. Consistent with this notion, indirect immunofluorescence of transient heterokaryons revealed that uninfected wild-type HTC cells failed to complement the defect in CR5 while CR5 cells did not suppress the wild-type phenotype of M1.54 cells. PMID- 2545482 TI - The action of hypothalamic and placental corticotropin releasing factor on the corticotrope. PMID- 2545483 TI - Altered adenylate cyclase activity in human otosclerotic bone cell cultures. AB - Adenylate cyclase (AC) activity was studied in whole homogenates of normal and otosclerotic bone cell cultures. When Mn2+ or Ca2+ was added to the medium there was a similar increase in AC activity in both cell types. F- provoked a greater rise in normal than in pathological cells, whereas 0.01 mM guanosine triphosphate (GTP) significantly raised cAMP synthesis in otosclerotic cells only. Mn2+ + calcitonin (Ct) increased AC activity in both cell preparations. With Ca2+ as cofactor there was no significant rise in either normal or pathological cells. However, while the combination Ca2+ + Ct + GTP had little effect on normal cells, it markedly increased cAMP synthesis in the pathological cells. 1 microgram/ml of the beta-blocker propranolol inhibited the effect Ct exerts on AC in normal cells, but enhanced it in otosclerotic cells. It would, therefore, seem that the pathogenesis of otosclerosis could be associated with an alteration in the AC system associated with Ct receptors. PMID- 2545484 TI - Cyclic AMP-responsive region of the human proopiomelanocortin (POMC) gene. AB - Transcription of the human proopiomelanocortin (POMC) gene is regulated by cAMP. To identify the region in the human POMC gene responsible for this regulation, we constructed chimeric genes containing different portions of the 5'-flanking region of the human POMC gene fused to the structural sequence encoding the bacterial reporter enzyme chloramphenicol acetyltransferase (CAT). The transcriptional activity of the fusion genes introduced into the rat glial cell line C6 was assayed by measuring CAT activity in the cell lysate. Forskolin, an adenylate cyclase-activating agent, stimulated the expression of POMC-CAT fusion genes. Deletion analysis demonstrated that the region between -417 and -97 bp from the transcriptional origin of the human POMC gene was responsible for regulation by cyclic AMP. PMID- 2545485 TI - Prolactin regulation of mouse mammary tumor virus (MMTV) expression in normal mouse mammary epithelium. AB - The hormonal regulation of mouse mammary tumor virus (MMTV) RNA in normal mouse mammary epithelium was studied in an explant system. In tissue from parous mice, physiological concentrations of prolactin stimulated MMTV expression, while only pharmacological concentrations of cortisol were effective. Regulation in explants from virgin mice was similar to that in parous animals except that the former were less sensitive to prolactin; this relative unresponsiveness may explain why uninduced tissue from virgin mice does not express MMTV RNA, while that from parous mice does exhibit some basal production. These results suggest that prolactin plays a major role in MMTV expression in normal mammary epithelium and that glucocorticoids may only have a permissive effect or may act through an indirect mechanism requiring high concentrations. These data also suggest that the greater susceptibility of parous mice to MMTV-induced tumorigenesis may reflect the greater prolactin sensitivity in the glands from these animals. PMID- 2545486 TI - Immunocytochemical localization, binding, and effects of atrial natriuretic peptide in rat adipocytes. AB - The metabolic effects of atrial natriuretic peptide (ANP) have not been widely investigated. Since adipocyte cells represent a model system extensively used to examine the metabolic actions of many peptide hormones, we sought to establish whether ANP could bind to adipocyte membranes, alter cyclic nucleotide metabolism, and affect spontaneous or hormone-stimulated lipolysis. Using in vitro autoradiographic techniques, radiolabelled ANP was found to bind specifically to mammary gland fat cells. Additionally, endogenous ANP-like immunoreactivity could be localized in the plasma membrane compartment and cytoplasmic matrix of fat cells, but not in fat vacuoles. [125I]ANP bound to single high affinity sites (Kd = 0.72 nM) in fat cell membranes. The binding was rapid (equilibrium within 1 min at 25 degrees C) and specific. The atrial peptide was capable of stimulating a time- and concentration-dependent increase in cGMP accumulation in isolated adipocytes, but had no effect on spontaneous or stimulated [-)-isoproterenol, ACTH, forskolin) cAMP formation. ANP did not alter the increase in glycerol production stimulated by l-epinephrine in isolated fat cells. While i.v. infusion of ANP stimulated a marked increase in circulating levels of cGMP, the atrial peptide did not alter plasma triglyceride levels. These data demonstrate the presence of specific ANP binding sites on adipocyte membranes and internalization of ANP-associated immunoreactivity. These receptors are biochemically functional given the ability of ANP to augment cGMP formation. The peptide, however, does not exert an action on adipocyte lipolysis. Adipocytes, therefore, represent an ANP target tissue in which the physiological action of the peptide is yet to be defined. PMID- 2545487 TI - Differential effects of transforming growth factor beta on human prostate cancer cells in vitro. AB - Transforming growth factor beta (TGF beta) exerts a wide spectrum of activity on many different cell types. Since TGF beta inhibits the growth of a variety of epithelial tumor cells in vitro, we examined the effects of TGF beta on the human prostate cancer cell lines DU145, PC3 and LNCaP for possible inhibitory activity. Growth in monolayer was initially inhibited in a dose-response fashion in the two androgen-independent cell lines, PC3 and DU145 but not in the androgen-dependent LNCaP cells. The rate of growth of the PC3 and DU145 cells treated with TGF beta, however, eventually returned to control levels despite retreatment with TGF beta. Anchorage-independent growth was inhibited to 55% and 16% control levels in PC3 and DU145, respectively. Scatchard analysis showed 1500 and 2900 TGF beta binding sites/cell on DU145 and PC3 cells with Kd = 6.9 and 12 x 10(-12) M, respectively. High-affinity binding could not be demonstrated on LNCaP cells. We also explored the possibility that TGF beta was secreted by these cells. Analysis of conditioned media by immunoprecipitation and a radioreceptor assay showed secretion of TGF beta into the media by DU145 and PC3 but not by LNCaP. Northern analysis showed the presence of TGF beta mRNA in DU145 and PC3, but not in LNCaP. These data indicate that TGF beta might serve as an autocrine inhibitory factor in prostate cancer. In addition, because TGF beta affects a wide range of cell types, TGF beta production by prostate cancer cells may contribute an important paracrine function in the development of tumor stromal tissue and metastases. PMID- 2545488 TI - Pertussis toxin enhances follicle-stimulating hormone-stimulated cAMP production in rat seminiferous tubules in a stage-dependent manner. AB - Cyclic (c) AMP production was measured in vitro in dissected pieces of adult rat seminiferous tubules of defined stages of the seminiferous epithelial cycle (VII VIII and XIV-IV) in the absence and presence of human follicle-stimulating hormone (FSH) (Metrodin, Serono). The basal rate of cAMP production was stage dependent, being about 2-fold higher in stages XIV-IV. FSH stimulated cAMP output in a time- and dose-dependent (stimulation at doses greater than or equal to 3 mg/l) fashion, and also the stimulability of stages XIV-IV (on average 2-fold) was greater than that of stages VII-VIII. When the tubular pieces were incubated in the presence of pertussis toxin (PT, 100 micrograms/l), the FSH-stimulated cAMP production of stages VII-VIII was enhanced by about 2-fold (P less than 0.01) whereas the basal rate was unaffected. In contrast, neither the basal nor the FSH-stimulated cAMP production of stages XIV-IV were affected by PT. Presence of the Gi-protein in both stages studied was demonstrated with PT-induced ADP ribosylation. However, no release of a putative activator of the Gi-protein could be demonstrated into spent media of the seminiferous tubules when incubated with freshly separated tubules. It is concluded that the poor FSH stimulability in cAMP production of certain spermatogenic stages of adult seminiferous tubules is at least partly due to endogenous inhibitors acting through the inhibitory Gi protein. This inhibition could be demonstrated in a stage-dependent manner, and was present in stages with the lowest production and least stimulability of cAMP production by FSH. PMID- 2545489 TI - Protein kinase A and C activities and endogenous substrates in ovine small and large luteal cells. AB - Protein kinase A (cAMP-dependent) and C (calcium, phospholipid-dependent) activities were measured and in vitro phosphorylation of endogenous proteins by these kinases were observed by SDS-PAGE in 100,000 x g supernatant (soluble) fractions of ovine small (12-22 microns) and large (greater than 22 microns) luteal cells. No differences in stimulation (P less than 0.05) of A kinase activity between small and large cells were detected. Protein kinase C activity was stimulated (P less than 0.05) 2.9-fold in small cells but not significantly enhanced above basal (P greater than 0.05) in large cells. By direct comparison, greater stimulation (P less than 0.05) over basal of A versus C kinase (6.1- versus 2.9-fold) was measured in small cells. These stimulations were greater than those observed in large cells (A kinase, 4.8-fold; C kinase, 1.8-fold). Maximal specific activities of both kinases (per mg protein) were greater (P less than 0.05) in small than in large cells. Endogenous proteins that could serve as substrates for phosphorylation by A and C kinases differed between small and large cells. Phosphorylation of six proteins by A kinase was consistently greater in small than in large cells. One endogenous protein (37 kDa) appeared to serve as a preferred substrate for phosphorylation by A kinase in small cells and C kinase in large cells. One protein (81 kDa) was predominantly phosphorylated in large rather than small cells by a calcium-dependent, C kinase-independent mechanism. These results support the accepted role of cAMP via A kinase and a possible role for C kinase in regulating steroidogenesis in ovine small luteal cells. The inability of large cells to respond to cAMP with enhanced secretion of progesterone may be due to an unavailability of phosphoprotein substrates for A kinase. Furthermore, protein kinase C activity and available protein substrates display quantitative and qualitative differences between small and large cells. Differences in regulation of steroidogenesis between the cell types may be due to these observed differences. PMID- 2545490 TI - Enhanced activity of the cardiac endocrine system during right ventricular hypertrophy. AB - In a model of pulmonary hypertension induced by a single injection of monocrotaline (MCT), we observed a time-dependent right ventricular hypertrophy, which became apparent in treated rats 21 days after administration of MCT and progressed through day 45. Associated with this right ventricular hypertrophy were time-dependent increases in ventricular levels of immunoreactive atrial natriuretic peptide (iANP). Forty-five days after MCT treatment, treated rats exhibited a 72-fold increase in right ventricular iANP levels and a 7-fold increase in left ventricular iANP levels. Hybridization analysis of total RNA extracted from cardiac tissue indicated that both atrial and ventricular ANP mRNA levels were elevated in treated rats. These data suggest that during pulmonary hypertension and cardiac hypertrophy the endocrine activity of the heart expands to include ventricular tissue. ANP binding site autoradiography revealed decreased binding site density in the kidney and hearts of treated rats at 49 days, consistent with the occurrence of desensitization/down-regulation. Enhanced ventricular ANP production may serve as a compensatory response to sustained elevation of pulmonary arterial pressure or may function as an autocrine/paracrine system regulating cardiac function. In either case, the effects of augmented ANP production may be subject to modulation by the status of ANP receptors in target organs and cells. PMID- 2545491 TI - Modulation of renin synthesis by lipoxygenase products in cultured human mesangial cells. AB - Modulation of renin synthesis by lipoxygenase products has been studied in cultured human mesangial cells under basal conditions and in the presence of prostaglandin (PG) E2. Total renin and cyclic AMP productions were stimulated in a dose-dependent manner (0.1-10 microM) by PGE2. The stimulatory effect of PGE2 on renin production was inhibited by 12-hydroxyeicosatetraenoic acid (12-HETE) between 0.1 and 100 nM. Extracellular and intracellular renin were affected similarly. Neither basal and PGE2-dependent cyclic AMP nor basal cyclic GMP productions were modified. 15-Hydroxyeicosatetraenoic acid (15-HPETE), 12 hydroperoxyeicosatetraenoic acid (12-HPETE) and 15-hydroperoxyeicosatetraenoic acid (15-HPETE) had the same effects as 12-HETE. Intracellular calcium concentration was not modified in the presence of 12-HETE. Since oleyl-2 acetylglycerol (OAG), an analog of diacylglycerol, also inhibited PGE2-stimulated renin production, it is hypothesized that the effect of the lipoxygenase products is mediated via protein kinase C stimulation. PMID- 2545492 TI - Establishment of 8-azaguanine-resistant mutants from rat thyroid cell line FRTL 5. AB - 8-Azaguanine (AG)-resistant mutant clones were isolated from FRTL-5 rat thyroid cells, by treating them with ethylmethane sulfonate for 48 h and then, after the recovery period, by selecting them in AG-containing medium. Isolated mutants were further selected in 6-thioguanine-containing medium, and two mutant clones were finally obtained (AG1 and AG2). The mutants retained several thyroid stimulating hormone (TSH) responsiveneses following acquisition of AG resistance: they responded to bovine TSH (bTSH), by producing cyclic 3',5'-adenosine monophosphate (cAMP) and thyroglobulin, and taking up iodine. However, TSH-responsive profiles were slightly different from each other in several points. They were killed in hypoxanthine-aminopterin-thymidine (HAT)-selective medium and applied to cell hybridization study with human thyrocyte from a Graves' patient: the hybrid turned out to produce both rat and human thyroglobulin even 12 months after the hybridization. PMID- 2545493 TI - Effect of pectin, a soluble dietary fiber, on functional and morphological parameters of the small intestine in rats. AB - We investigated the effects of pectin, a soluble dietary fiber, on functional and morphological parameters of the small intestine in rats. A control group and a pectin-fed group were given a fiber-free elemental liquid diet and an elemental liquid diet containing 2.5% (w/w) pectin, respectively, for 2 weeks. The ileal mucosal specific activities of maltase, sucrase and alkaline phosphatase increased significantly in the pectin-fed group. Maltose absorption of the ileum, studied in vitro by the method of everted sacs and disaccharide-dependent potential difference, increased significantly in the pectin-fed group. The length of the small intestine as well as the villus height and crypt depth of both the jejunum and the ileum were significantly greater in the pectin-fed group. The crypt cell production rate of the jejunum and the ileum was also significantly greater in the pectin-fed group. Plasma enteroglucagon, but not gastrin, increased significantly in the pectin-fed group. These data suggest that pectin feeding results in hyperplasia of the small-intestinal mucosa and a significant increase in the enzyme activities of the brush border membrane of the ileum. PMID- 2545494 TI - In vitro study of effects of 1,25 dihydroxyvitamin D3 on the morphology of human breast cancer cell line BT.20. AB - The morphological effect induced by 1,25 dihydroxycholecalciferol - 1,25(OH)2VitD3 - on the malignant human breast cell line BT.20 was studied. This cell line is devoid of oestrogen (ER) and progesterone (PGR) receptors. This effect, which requires treatment for at least 3 days, was evidenced by an increase in the cell projection surface, assessed on scanning electron microscopy (SEM) and by quantimetric analysis, for optimal final concentrations of 1.25(OH)2VitD3 in the medium of the order of 10(-11)M. The cells became more spread out and rounded with many junctional systems; there was occlusion of the intercellular space, but hardly any cells overlapped. The cytoskeleton was considerably developed, with microtubules running parallel to the cell surfaces associated with microfilaments. This positive action on cell differentiation was very similar to that noted regarding the reduction in growth under 1,25(OH)2VitD3 treatment. The two actions were, however, observed at different times and with different concentrations of 1,25(OH)2VitD3 in the medium. PMID- 2545495 TI - Endothelium-derived relaxing and contracting factors. AB - Endothelium-dependent relaxation of blood vessels is produced by a large number of agents (e.g., acetylcholine, ATP and ADP, substance P, bradykinin, histamine, thrombin, serotonin). With some agents, relaxation may be limited to certain species and/or blood vessels. Relaxation results from release of a very labile non-prostanoid endothelium-derived relaxing factor (EDRF) or factors. EDRF stimulates guanylate cyclase of the vascular smooth muscle, with the resulting increase in cyclic GMP activating relaxation. EDRF is rapidly inactivated by hemoglobin and superoxide. There is strong evidence that EDRF from many blood vessels and from cultured endothelial cells is nitric oxide (NO) and that its precursor is L-arginine. There is evidence for other relaxing factors, including an endothelium-derived hyperpolarizing factor in some vessels. Flow-induced shear stress also stimulates EDRF release. Endothelium-dependent relaxation occurs in resistance vessels as well as in larger arteries, and is generally more pronounced in arteries than veins. EDRF also inhibits platelet aggregation and adhesion to the blood vessel wall. Endothelium-derived contracting factors appear to be responsible for endothelium-dependent contractions produced by arachidonic acid and hypoxia in isolated systemic vessels and by certain agents and by rapid stretch in isolated cerebral vessels. In all such experiments, the endothelium derived contracting factor appears to be some product or by-product of cyclooxygenase activity. Recently, endothelial cells in culture have been found to synthesize a peptide, endothelin, which is an extremely potent vasoconstrictor. The possible physiological roles and pathophysiological significance of endothelium-derived relaxing and contracting factors are briefly discussed. PMID- 2545496 TI - Antagonists of B2 bradykinin receptors. AB - Bradykinin and its active metabolites, produced by kallikreins at their sites of action, potently elicit a variety of biological effects: hypotension, bronchoconstriction, gut and uterine contraction, epithelial secretion in airway, gut, and exocrine glands, vascular permeability, pain, connective tissue proliferation, cytokine release, and eicosanoid formation. These effects are mediated by at least two broad classes of receptors. The most common is the B2 subtype. The availability of competitive antagonists of B2 receptors has provided powerful tools for the study of bradykinin's actions. The significance of kinins in certain human diseases is being explored by using these agents as potential therapeutic agents. Human clinical trials are under way to test the usefulness of bradykinin receptor antagonists to treat symptoms of the common cold and the pain associated with severe burns. Trials are also being comtemplated for use in treatment of asthma. PMID- 2545497 TI - Cell-free heterologous desensitization of adenylyl cyclase in S49 lymphoma cell membranes mediated by cAMP-dependent protein kinase. AB - We have examined the cell-free heterologous desensitization of adenylyl cyclase in plasma membrane preparations from S49 wild-type (WT) and kin- cells (which lack cAMP-dependent protein kinase) incubated with purified catalytic subunit of cAMP-dependent protein kinase (cA.PKc). cA.PKc caused a rapid (t1/2 = 40 s) decrease in the hormone responsiveness of adenylyl cyclase in the WT membrane preparations that mimicked the intact cell heterologous desensitization; that is, there was an increase in the Kact for both epinephrine and prostaglandin E1 (PGE1) stimulations of adenylyl cyclase induced at the receptor level because neither forskolin- nor NaF-stimulated activity was affected. The desensitization was independent of agonist occupancy of the receptor, and the effects were blocked both by the active fragment (amino acids 5-22) of the specific inhibitor of cA.PK and by p[NH]ppA. cA.PKc treatment of kin- membranes resulted in a heterologous desensitization that resembled the effects of WT adenylyl cyclase, with the exception that forskolin-stimulated activity was also reproducibly decreased by 24%. cA.PKc had no effect on WT membranes isolated from cells that had previously undergone maximal heterologous desensitization during treatment with 10 microM forskolin. In contrast, cA.PKc-induced heterologous desensitization of kin- membranes was additive with the epinephrine-induced homologous desensitization of intact cells. Cell-free desensitizations were reversed by incubation of membranes with cA.PKc and ADP, conditions that drive the kinase reaction backward. The similarities of our cell-free cA.PKc-mediated heterologous desensitization of adenylyl cyclase with the intact cell desensitization support our hypothesis that heterologous desensitization of the WT lymphoma cells is mediated by cA.PK via a mechanism independent of homologous desensitization. PMID- 2545498 TI - Age and sex differences in bone metastasis of hepatocellular carcinoma in Japanese autopsy cases. AB - Metastatic patterns of hepatocellular carcinoma (HCC) in 6997 autopsy cases recorded in the Annual of the Pathological Autopsy Cases in Japan from 1981 to 1984 were analyzed by using a computer. Significant association of the metastatic pattern of hepatocellular carcinoma with age and sex was shown after separating patients into different age groups in pack years. It was especially worthy of notice that bone metastasis decreased in an age-dependent manner only in men within the widest age range (P less than 0.001) and was significantly prevalent in men in their forties and fifties (P = 0.012). In addition, in 350 cases of hepatocellular carcinoma autopsied at Tokyo University Hospital during 15 years from 1971 to 1985, it was suggested that bone metastasis might be prevalent in male patients with liver cirrhosis although significant relationship between bone metastasis and hepatitis B virus infection or alcohol abuse was not shown. These results might indicate possible effects of sex differences on the metastatic pattern of hepatocellular carcinoma, particularly bone metastasis, as well as on the incidence of the cancer. PMID- 2545499 TI - Antipyrine clearance per unit liver volume in cirrhotics with and without hepatocellular carcinoma indicating a correlation with histological change of the liver. AB - The antipyrine metabolizing capacity was studied in 12 patients with cirrhosis of the liver and 12 with cirrhosis and hepatocellular carcinoma (HCC). Antipyrine clearance (Cl) and liver volume (LV) were measured and the antipyrine clearance per unit liver volume (Cl/LV) was calculated. The patients with HCC showed a significantly lower Cl value than those without HCC but there was no significant difference in Cl/LV between the two groups. This suggested that the lower Cl values in the HCC patients resulted from a decrease in residual liver mass. Cl/LV showed positive correlation with % parenchymal cell mass as an indicator of residual parenchymal cell mass per unit volume of liver. This result showed a correlation of Cl/LV with histological change of the liver in cirrhotics. PMID- 2545500 TI - Decreased membrane fluidity in erythrocytes from patients with Crohn's disease. AB - In erythrocytes from patients with Crohn's disease, lower levels of membrane fluidity were demonstrated by means of electron spin resonance with 16-stearic acid spin label (SAL) but not 5- or 12-SAL, suggesting more apparent abnormality in the deeper portion of lipid bilayer. Membrane lipid analysis showed a significant increase of sphingomyelin, a decrease of phosphatidylcholine and decreased contents of polyunsaturated acyl-chains, particularly C18:2, of phospholipid, without alteration in the cholesterol to phospholipid molar ratio. The alteration in membrane fluidity appeared to be closely related to changes in membrane phospholipid classes and their acyl-chains. Among the plasma lipids, contents of esterified cholesterol, phospholipid, very low- and low-density lipoproteins and cholesterol of high-density lipoprotein significantly decreased. Decreased contents of polyunsaturated acyl-chains, particularly C18:2, in the plasma phospholipid were also noted. Changes in plasma and erythrocyte membrane lipids remained even in the inactive stage of the disease, except for the normalized sphingomyelin and phosphatidylcholine content in the erythrocyte membrane. Since membrane fluidity is closely involved in various cell membrane functions, the decreased membrane fluidity and altered lipid composition in erythrocyte membrane evident in patients with Crohn's disease may have important effects on the pathological status in this disease. PMID- 2545501 TI - Non-septic endotoxemia in cirrhotic patients. AB - We have found that endotoxemia detected by conventional LCT (limulus colorimetric test) in patients with liver diseases could not be detected by endotoxin-specific LCT at all, and proposed that this beta-glucan like activity (BGLA) should be termed as non-septic endotoxemia, distinguishing it from septic endotoxemia seen in gram-negative sepsis. In this study, we investigated non-septic endotoxemia through the clinical course of 8 cirrhotic patients. Non-septic endotoxemia appeared at the onset of DIC but tended to decline in level in the late terminal stage. This phenomenon cannot be consistent with the "spillover" theory which explains the mechanism of endotoxemia without sepsis in liver disease. We think it is an urgent problem to elucidate the nature of BGLA in liver disease, without recourse to the "spillover" theory. PMID- 2545502 TI - Hypomethylation of the c-myc oncogene in liver cirrhosis and chronic hepatitis. AB - In order to investigate how chronic liver diseases, including liver cirrhosis and chronic hepatitis, are associate with hepatocarcinogenesis in terms of gene alteration, the methylation states of the c-myc and c-Ki-ras genes were examined in 34 liver tissues from patients with chronic liver disease without hepatocellular carcinoma (HCC), 34 non-tumor liver tissues from patients with HCC, 18 HCC tissues and 31 control liver tissues. The methylation states were analyzed by the Southern hybridization method using the restriction endonuclease isoschizomers MspI and HpaII. The CCGG sites at the second exon of the c-myc gene tended to be more extensively hypomethylated both in chronic liver disease and in non-tumor tissues than in control livers. Whereas the CCGG sites of the c-Ki-ras, and the third exon of the c-myc gene tended to be hypomethylated only in HCC tissues in comparison with other tissue groups. These results suggest that chronic liver disease may be situated between normal liver and HCC based on the state of DNA methylation and associated with the development of HCC through hypomethylation of the c-myc and/or c-Ki-ras gene. PMID- 2545503 TI - A case report of transcatheter arterial embolization of cholestatic type of hepatoma. AB - A 57-year-old man with hepatocellular carcinoma (HCC) invading the hepatic duct was treated with transcatheter arterial embolization (TAE). The dilated hepatic duct was decompressed his jaundice disappeared and he survived for 10 months after the first TAE, succumbing due to bleeding from gastric erosion. Although total bilirubin was 26.7 mg/dl and massive ascites was noted on occasion of the first TAE, hepatic dysfunction did not worsen and both icterus and ascites decreased, disappearing one month after the therapy. Autopsy revealed HCC with trabecular arrangement originating in the left lobe and growing into the left hepatic duct accompanied by liver cirrhosis and follicular carcinoma of the thyroid gland. Severe icterus caused by bile duct obstruction does not correlate with the grade of hepatic failure in HCC with liver cirrhosis, so TAE was effective and should be tried as a first choice therapy in such a poor risk case. PMID- 2545504 TI - Inflammatory mediators of experimental colitis in rats. AB - Colonic inflammation was induced in rats by intracolonic administration of 0.25 ml of 50% ethanol containing 30 mg of trinitrobenzene sulfonic acid (TNB). Control rats were treated with 0.25 ml of 50% ethanol or with 30 mg of TNB in 0.25 ml of saline. After 24 h, mucosal ulceration and hemorrhage were observed in TNB/ethanol-, 50% ethanol-, and to a lesser extent, in TNB/saline-treated rats. After 1 wk, mucosal damage was completely resolved in the 50% ethanol and TNB/saline-treated rats but the lesions in the TNB/ethanol-treated rats persisted and progressed to a chronic active inflammatory process after 3 wk. Myeloperoxidase activity was significantly elevated in mucosal scrapings from all treatment groups at all time intervals when macroscopic and microscopic mucosal injury was evident. Interleukin-1 was found to be the most sensitive indicator of mucosal inflammation, and its mucosal values correlated with myeloperoxidase activity. Leukotriene B4 was increased in control rats at 1 wk and in TNB/ethanol treated rats at all time intervals. The maximal increase in leukotriene B4 was observed at 1 wk. Thromboxane B2 generation was reduced while platelet activating factor generation was not increased in TNB/ethanol-treated rats. These results indicate that in this TNB/ethanol model of gut inflammation, myeloperoxidase activity and interleukin-1 are reliable and sensitive indicators of colonic inflammation, and that thromboxane B2 is not involved in the acute lesions, whereas leukotriene B4 appears in the chronic active inflammatory response. PMID- 2545505 TI - Structural and enzymatic changes during colonic maturation in the fetal and suckling rat. AB - To assess correlations between cellular differentiation and enzymatic maturation in the developing rat colon, tissue from fetal, suckling, weanling, and adult rats was analyzed by electron microscopy and assayed for lactase, alkaline phosphatase, and sodium-potassium-stimulated adenosine triphosphatase activities. The proximal and distal colon were analyzed independently at all ages. All three enzymes were detected in the fetal colon when the cells were highly undifferentiated. Postnatally, significant regional differences in cellular ultrastructure appeared, only some of which were directly paralleled by enzymatic changes. Each enzyme had a distinct region-specific developmental pattern. Lactase and sodium-potassium-stimulated adenosine triphosphatase were significantly enhanced at birth, decreasing to adult levels by 15 days postnatal. Regional differences were present, but the patterns were similar. These patterns did not parallel the increase in microvillar height and number and basolateral interdigitations of the surface columnar cells, the structural correlates of lactase, and sodium-potassium-stimulated adenosine triphosphatase, respectively. In contrast, developmental changes in alkaline phosphatase activity paralleled structural maturation, at least in part. The activity levels in the distal colon did not change significantly with age and few major structural changes were noted. In the proximal colon, activity increased markedly after birth, and after 10 days decreased rapidly to adult levels, a pattern that coincided with the transient appearance of villi and specialized cells with apical tubules and vesicles known to have alkaline phosphatase activity. The results show age- and region-related changes in cellular ultrastructure and enzymatic activities, only some of which appear to be directly correlated. PMID- 2545506 TI - Short- and long-term effects of reserpine on rat cardiac beta-adrenoceptors. AB - 1. In order to explore the mechanism of reserpine-induced supersensitivity, we examined the effects of various doses of reserpine (R) on rat cardiac beta adrenoceptors (beta AR). 2. 24 hr after a single administration of R (1 mg/kg, i.p.) beta AR were up-regulated, while with higher doses (2.5-5 mg/kg) beta AR density was not changed but beta AR coupling, evaluated by the proportion of high affinity agonist binding states, was impaired. 3. Long-term effects of R (2.5 mg/kg/day, 2 days) included beta AR down-regulation and uncoupling 24 hr treatment (day 1), followed by up-regulation at day 7, while all parameters were similar to controls at day 14. 4. It appears that R induces either an increase or a decrease of cardiac beta AR density and coupling, dependent on the dose and the time at which beta AR are measured after treatment. PMID- 2545507 TI - Modulation of noradrenaline release from cat cerebral arteries by presynaptic alpha 2-adrenoceptors. Effect of chronic treatment with desipramine and cocaine. AB - 1. Field electrical stimulation elicited an increase of the tritium efflux over the basal level from cat cerebral arteries previously incubated with (+/-) [3H]noradrenaline ([3H]NA). 2. This efflux was: (a) reduced by clonidine, NA or B HT 920; (b) unaffected by methoxamine, prazosin and yohimbine (10(-6) M); (c) reduced by yohimbine (5 x 10(-6) M), and (d) increased by phentolamine. 3. The effect of clonidine was blocked by yohimbine. 4. The daily treatment with the neuronal uptake blockers desipramine (10 mg/kg, i.p.) or cocaine (10 mg/kg, i.p.) [during 12 days], antagonized the inhibitory action of clonidine totally or partially, respectively. 5. These results suggest: (1) the existence of presynaptic alpha 2-adrenoceptors in these arteries, which modulate the NA release, and (2) that chronic treatment with desipramine or cocaine induces a subsensitivity of these alpha 2-receptors, which facilitates the NA release. PMID- 2545508 TI - Cyanide intoxication--I. An oral chronic animal model. AB - The effects of oral chronic cyanide administration to mice were studied. Cyanide intoxication was confirmed by the increased levels of this poison and the concomitant inhibition of cytochrome oxidase activity in liver, brain, heart and blood. The detoxifying enzyme rhodanese was measured. The state of the sulfane sulfur pool was investigated by determination of the cyanide labile-sulfur levels. A clear correlation between rhodanese activity and sulfur content was obtained as a consequence of cyanide action. These results support the belief that rhodanese plays a fundamental role in the detoxification process of cyanide, in preventing cyanide reaching the target tissues. PMID- 2545509 TI - Effects of non-steroidal anti-inflammatory drugs on isolated human polymorphonuclear leukocytes (PMN): chemotaxis, superoxide production, degranulation and N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP) receptor binding. AB - 1. We have examined the effects of tolmetin and meclofenamate on isolated human PMN functions under FMLP stimulating conditions. 2. In a dose dependent manner, tolmetin and meclofenamate inhibited all PMN functions, except that tolmetin stimulated PMN chemotaxis. 3. Meclofenamate was much more potent than tolmetin as an inhibitory agent. 4. We also conducted competitive receptor binding assays for tolmetin, meclofenamate and ibuprofen on the FMLP receptor. 5. All three NSAID inhibited FMLP binding in a dose dependent manner with the potency order being meclofenamate greater than ibuprofen greater than tolmetin. PMID- 2545510 TI - Diazepam effects on frog skin cyclic nucleotide phosphodiesterase. AB - 1. The activities of cAMP phosphodiesterases (cAMP-PDE) have been measured in the homogenate of the skin of Rana esculenta. 2. The tissue possesses two distinct enzymes: a "low" Km PDE (Km = 0.42 x 10(-6) M; Vmax = 16 pmol/mg protein/min) and a "high" Km PDE (Km = 180 x 10(-6) M; Vmax = 2853 pmol/mg prot/min). Only the "high" Km form is stimulated by calcium. 3. Diazepam (1-0.5 mM) significantly inhibits both enzymes, the inhibition being of competitive type. PMID- 2545511 TI - Amphetamine antagonizes the presynaptic inhibitory effect of clonidine through an interaction at the level of the alpha 2-adrenoceptors. AB - 1. In the whole rat vas deferens 20 microM noradrenaline (NA) and 0.011 microM clonidine decreased (36 +/- 7.4% and 80 +/- 6.0% respectively) the motor response induced by hypogastric nerve stimulation. These effects were reverted by 1 microM yohimbine. Amphetamine 5.4 microM failed to antagonize the inhibitory effect of NA and attenuated clonidine effect. 2. The effect of amphetamine was not altered by preincubation with either cocaine 1 microM, (-)-propranolol 0.3 microM or cocaine plus prazosin 0.028 microM. 3. In reserpine pretreated animals amphetamine 5.4 microM shifted to the right the concentration-response curve (CRC) to clonidine 0.62 +/- 0.05 log units with a KB value of 1.83 +/- 0.30 microM. 4. Binding of [3H]clonidine and [3H]prazosin were inhibited by amphetamine. Amphetamine was 90 times more potent to inhibit [3H]clonidine binding. 5. The results obtained suggest a possible direct interaction between clonidine and amphetamine on alpha-adrenoceptor. PMID- 2545512 TI - Selective potentiation of extracellular Ca2+-dependent contraction by neuropeptide Y in rabbit mesenteric arteries. AB - 1. Neuropeptide Y (NPY) potentiated the contractile responses induced by electrical transmural stimulation, noradrenaline and KCl in the rabbit mesenteric artery. 2. In preparations treated with noradrenaline or KCl in Ca2+ free medium, NPY also potentiated the contractile response induced by resupplementation of Ca2+. 3. 3H-efflux from the arteries preincubated with [3H]-noradrenaline was not affected by NPY. 4. These results suggest that NPY selectively acts on the postsynaptic membrane and potentiates the contractions mediated through receptor operated and voltage-dependent Ca channels. PMID- 2545513 TI - Smoltification and seawater adaptation in coho salmon (Oncorhynchus kisutch): plasma calcium regulation, osmoregulation, and calcitonin. AB - In order to examine the dynamics of ion regulation, osmoregulation, and plasma calcitonin during the parr-smolt transformation (smoltification), blood and gill tissue were collected from yearling coho salmon, Oncorhynchus kisutch, from February to October. Fish were kept in fresh water (FW) throughout this period. In addition, fish were exposed to seawater (SW) at the peak of smoltification in mid-April, and samples from these fish were collected until July. Plasma osmolality, gill Na+,K+-ATPase activity, plasma levels of calcitonin, and free and total calcium and magnesium were measured. SW adaptability of FW fish was assessed throughout the study by measurements of plasma osmolality following a 24 hr exposure to seawater. The greatest hypoosmoregulatory ability occurred in April-May, although SW-adapted fish had higher plasma osmolality than FW-adapted fish at all times. Gill Na+,K+-ATPase activity in FW-adapted fish increased from April to June and increased rapidly following exposure of fish to SW, and remained elevated in SW-adapted fish. Free plasma calcium and magnesium levels increased following SW exposure, but returned to prior levels within 1 week. Netting and confinement stress during sampling caused an increase in plasma osmolality and free calcium and magnesium levels in both FW- and SW-adapted fish. Changes in hypoosmoregulatory ability during smoltification and SW adaptation were correlated with changes in gill Na+,K+-ATPase activity. A sharp transitory peak in plasma calcitonin levels occurred early in smoltification (March) and in SW-adapted fish in June. Plasma calcitonin levels gradually increased in FW adapted fish during the period of desmoltification. However, no change in plasma calcitonin levels occurred during SW-induced hypercalcemia, suggesting that the hormone does not play a major role in short-term plasma calcium regulation in coho salmon. PMID- 2545514 TI - Smoltification and seawater adaptation in Atlantic salmon (Salmo salar): plasma prolactin, growth hormone, and thyroid hormones. AB - To obtain more information on the role of prolactin and growth hormone during the parr-smolt transformation of Atlantic salmon, a population of fish in fresh water was sampled from January to June during two consecutive years. Gill Na+,K+-ATPase activity increased steadily during smoltification and a plasma thyroxine peak was observed 2-3 weeks before the gill Na+,K+-ATPase peak. On the basis of these two parameters, smoltification was considered complete in our populations in April 1985 and May 1986. Two peaks in plasma growth hormone levels occurred in 1986, one in mid-April and the second in mid-May. In both cases, these peaks coincided with a peak in plasma triiodothyronine and preceded the thyroxine peak by 1-2 weeks. Moreover, the second peak which lasted for 1 month coincided with maximal gill Na+,K+-ATPase activity. A decrease in plasma prolactin levels was observed during smoltification of Atlantic salmon in 2 consecutive years. During this period of decreasing and low plasma prolactin levels, gill Na+,K+-ATPase activity increased to its highest values. Atlantic salmon smolts were also directly transferred into seawater. After 2 days or more in seawater, plasma prolactin levels were not significantly different from those on Day 0, whereas in fresh water they showed large fluctuations. All these data indicate that growth hormone may play an important role in the development of hypoosmoregulatory activity. Increased hypoosmoregulatory ability also appears to be associated with low prolactin levels. PMID- 2545515 TI - Alterations in pituitary GnRH and dopamine receptors associated with the seasonal variation and regulation of gonadotropin release in the goldfish (Carassius auratus). AB - Seasonal variations in the serum concentrations of gonadotropin (GtH) and the serum GtH response to intraperitoneal injection of domperidone, a specific dopamine receptor antagonist, were examined in goldfish. In addition, the effects of in vivo treatment of goldfish with a superactive analog of salmon gonadotropin releasing hormone (sGnRH-A) and domperidone on the binding parameters of pituitary GnRH and dopamine receptors were investigated in goldfish. Serum concentrations of GtH and the maximum GtH response to domperidone increased in correlation with advancing gonadal maturation; values increased from those in sexually regressed fish in January to maximal levels observed in fish in late stages of gonadal recrudescence in March, followed by a decrease with gonadal regression. At all stages, injection of domperidone increased serum concentrations of GtH in a dose-related manner; however, the ED50 of domperidone did not vary significantly over the course of the reproductive cycle. Multiple injections of sGnRH-A caused a progressively increasing and more prolonged serum GtH response; as well, multiple sGnRH-A treatment significantly potentiated the serum GtH response to domperidone without altering the ED50 of domperidone. sGnRH A treatment caused a significant increase in the number of dopamine/neuroleptic receptors in the goldfish pars distalis, accompanied by a nonsignificant increase in dopamine/neuroleptic receptors in the neurointermediate lobe, and significantly increased the number of high-affinity GnRH receptors in the goldfish pituitary. Treatment with domperidone also significantly increased pituitary high-affinity GnRH receptor numbers. Receptor affinities were not significantly altered by either sGnRH-A or domperidone treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545516 TI - Cloning of the glycerol kinase gene of Bacillus subtilis. AB - A 3.5 kb fragment of Bacillus subtilis DNA which contains wild type alleles of mutations in glpK (glycerol kinase) and glpD (glycerol-3-phosphate [G3P] dehydrogenase) was cloned in plasmid pHV32 in Escherichia coli. The cloned fragment expresses glycerol kinase in B. subtilis mutants carrying the mutations glpK11 and recE4 after induction with glycerol or G3P whereas it does not express G3P dehydrogenase. The cloned fragment thus contains the complete glpK but probably only part of glpD. PMID- 2545517 TI - Transformation of Bacillus thuringiensis by electroporation. AB - Plasmids were transformed by electroporation into various strains of Bacillus thuringiensis with frequencies of up to 10(5) transformants/micrograms. pC 194 transformed all strains tested at a high frequency and cells could be stably transformed with pC194 and pUB110 simultaneously by electroporation with a frequency of 10(2) pC194+ pUB110 transformants/micrograms DNA. Low transformation frequencies observed with some plasmids, especially those grown initially in Escherichia coli, could be increased by passage through B. thuringiensis, B. thuringiensis var. israelensis and in acrystalliferous mutant of the same strain transformed at frequencies of 10(4)-10(5)/micrograms DNA with most of the plasmids tested. A cloned israelensis 27-kDa delta-endotoxin gene was introduced into the israelensis acrystalliferous mutant and a kurstaki acrystalliferous mutant by electroporation. Both transformants were shown to express the endotoxin gene and to be toxic to Aedes aegypti larvae. PMID- 2545518 TI - Contraction induced by Clostridium perfringens epsilon toxin in the isolated rat ileum. AB - Clostridium perfringens epsilon toxin caused contraction of the isolated ileum of the rat in a dose-dependent manner. The contraction caused by the toxin was inhibited by a low Na medium, tetrodotoxin (TTX), atropine, mecamylamine or tetraethylammonium (TEA). Furthermore, the contractile response induced by the toxin was abolished by incubation in Ca-free medium, and completely restored by and addition of Ca2+. In addition, verapamil inhibited contraction induced by the toxin in a dose-dependent manner. These data suggest that epsilon toxin induces contraction of the isolated ileum and that the toxin-elicited contraction is the result of an indirect action mediated through the nervous systems. PMID- 2545519 TI - The cytochrome oxidases of Bacillus subtilis: mapping of a gene affecting cytochrome aa3 and its replacement by cytochrome o in a mutant strain. AB - A mutant of Bacillus subtilis has been isolated that fails to grow on succinate as the source of carbon, yet grows on glucose. Intact cells of the mutant and cytoplasmic membranes derived therefrom lack cytochromes a and a3 but contain a cytochrome o-like pigment, which forms a photodissociable compound with CO and is reactive with oxygen. The mutation in the genome has been located and lies at about 130 degrees on the chromosomal map between the metC and pyrD loci. The designation cox is suggested for this gene. PMID- 2545520 TI - [Interaction of products of su(Hw) and su(f) genes with MDG4 region regulating its transcription suppresses mutations in Drosophila caused by insertion of this gene]. AB - The mdg4 (gypsy) mobile element of Drosophila contains two closely situated regions binding to proteins from nuclear extracts. One of these is an imperfect palindrome having homology with the lac operator of Escherichia coli, the other contains a reiterated sequence (5'PyPuTCTGCATACTPyPy) homologous to the octamer which is the core of many enhancers and upstream promoter elements. The transient expression of deletion mutants has shown that these DNA regions are negative and positive regulators, respectively, of mdg4 transcription. As was demonstrated earlier, mutations induced by the presence of mdg4 at different loci are suppressed, owing to either repression or activation of mdg4 transcription in Drosophila lines carrying unlinked mutations in su(Hw) or su(f) genes. We have shown that binding to a negative regulator (silencer) is weakened in nuclear extracts isolated from cell lines carrying su(f) mutations which activate mdg4 transcription; therefore, the su(f) gene codes for a protein capable of mdg4 repression. Furthermore, binding to a positive regulator is weakened in nuclear extracts isolated from cell lines carrying su(Hw) gene mutations which decrease the level of mdg4 transcription; hence, the su(Hw) gene encodes a protein which activates mdg4 transcription. PMID- 2545521 TI - [Characteristics of Escherichia coli K-12 mutants with altered effectiveness of excision of Tn5 and Tn9 transposons]. AB - The properties of Escherichia coli K-12 mutans HFETn5, HFETn9 and LFETn9 have been studied. The majority of mutations were shown to have pleiotropic effect. Some of them increase cell sensitivity to UV light and mitomycin C and affect efficiency of homologous recombination in transduction and conjugation. The level of spontaneous mutagenesis is increased in a number of mutants. None of the mutations isolated affect frequency of transposition of Tn5 from bacteriophage lambda::Tn5 into the chromosome. Based on analysis of properties of hfeTn5-09 and hfeTn9 mutations and on the date of preliminary mapping of hfeTn5-09 mutation, these mutations were considered to be novel. It is shown that the processes of precise excision of Tn5 and Tn9 transposons may be accomplished by at least two pathways, one of them being dependent on recA gene functions. PMID- 2545522 TI - Molecular analysis of an alcohol dehydrogenase (Adh) gene from chromosome 1 of wheat. AB - We have cloned and determined the nucleotide sequence of a gene encoding alcohol dehydrogenase (Adh) from Triticum aestivum cv. Millewa. Southern analysis using cv. Chinese Spring nullisomic-tetrasomic and ditelosomic lines established that the cloned gene mapped to the long arm of chromosome 1A and does not correspond to any previously identified wheat Adh locus. Southern analysis also provided evidence for triplicate copies of this Adh gene on the homoeologous group 1 chromosomes, while Northern blots indicated that the homoeologous group 1 Adh genes, like several other plant Adh genes, are transcribed under anaerobic conditions. Sequence analysis indicates that the cloned gene has a structure similar to both monocot and dicot Adh genes with an open reading frame encoding a polypeptide of 379 amino acids. Sequences important for eucaryotic gene expression such as the TATA box, polyadenylation signal, and intron splice sites were found in the expected positions. The open reading frame is interrupted by 8 introns which are in identical positions with 8 of the 9 introns in maize and pea Adh genes, suggesting that during evolution there are processes occurring that result in the loss of introns. Sequence analysis also revealed that the cloned wheat Adh gene shared extensive homology with the barley Adh3 gene not only in the coding region but also in the noncoding regions. However, this homology is discontinuous as a result of a 1.8-kbp insertion (TLM), which is present in the cloned wheat Adh gene and absent in the barley Adh3 gene. Sequence analysis of this insertion reveals features characteristic of the short terminal inverted repeat class of eucaryotic transposable elements. We have no evidence for the transposition of the TLM element. However, Southern blots reveal multiple copies of sequences related to TLM in the wheat genome and in other closely related species, suggesting that transposition may once have played an important role in the evolution of the Gramineae family. PMID- 2545523 TI - A second locus for the 5S multigene family in Secale L.: sequence divergence in two lineages of the family. AB - The 5S RNA genes in Secale sp. are arranged as tandem arrays of a 460- and 480-bp repeating sequence. These size classes were initially discovered by restriction endonuclease analysis using BamHI and subsequently by DNA sequencing of cloned units. The length variation between short and long units originated from major deletion-insertion events in the noncoding spacer region of the 5S DNA repeat units. In situ hybridization with [3H]cRNA and biotin-labelled probes synthesized from both the short and long 5S DNA units of S. cereale localized the sites on chromosome 1R and a new site on a chromosome identified as 5R. We propose that the chromosome 1R locus, which has been mapped previously, be named 5SDna-R1 and the second locus, reported in the present paper, be referred to as 5SDna-R2. A preferential hybridization of a probe from the long unit to the 5SDna-R2 locus and of a probe from the short unit to the 5SDna-R1 locus is reported. The clustering of long units in the 5SDna-R2 locus was confirmed by restriction endonuclease digestion of DNA from rye chromosome 5R additions to wheat. Nucleotide sequence alignment of 5S DNA repeat units from a number of Secale species, using both phenetic and cladistic computer programmes, demonstrated that two clear lineages corresponding to the long and short units existed in this genus. The different Secale species could not be unambiguously differentiated using the 5S DNA sequences. PMID- 2545524 TI - CpG methylation of the cAMP-responsive enhancer/promoter sequence TGACGTCA abolishes specific factor binding as well as transcriptional activation. AB - In mammals and other vertebrates, cytosine methylation in CpG sites is often negatively correlated with gene activity. Because methylation of the promoter region is most crucial for this effect, the simplest hypothesis is that CpG methylation interferes with the binding of specific transcription factors. We have examined this hypothesis with two different transcription factor-binding sites that contain a CpG dinucleotide, namely the cAMP-responsive element (CRE; 5'-TGACGTCA) and the Sp1-binding site (5'-GTGAGGCGGTGAGACT). We have reported previously that CpG methylation of the Sp1-binding site affected neither factor binding nor transcription in HeLa cells, which may be related to the fact that Sp1 is typically associated with promoters of housekeeping genes. In contrast, CREs are often associated with promoters of cell type-specific genes. A synthetic oligonucleotide containing two tandem CREs derived from the gene encoding the human glycoprotein hormone alpha-subunit was cloned upstream of a reporter gene. Transcription of this gene was dependent on the CRE sequences in both PC12 and HeLa cells. Bandshift and methylation interference assays show that similar, if not the same, factor(s) bind to the CRE in both cell lines, even though induction by cAMP was only observed in PC12 cells. CpG methylation of the CRE consensus sequences (TGACGTCA) resulted in loss of specific factor binding, as well as loss of transcriptional activity in vitro and in vivo, in both cell types. This suggests that the inactivity of methylated promoters can, at least in some cases, be explained by their inability to bind specific transcription factors. PMID- 2545526 TI - Analysis of the cis-acting requirements for germ-line-specific splicing of the P element ORF2-ORF3 intron. AB - P-element transposition is limited to the germ line because the element's third intron is only spliced in germ line cells. We show that a 240-bp fragment containing this 190-bp intron can confer germ line specificity when placed in the context of another gene. We find that the cis-acting regulatory sequences required for germ line regulation map near to, but not at, the 5' or 3' splice junctions. PMID- 2545527 TI - cis-acting DNA sequence requirements for P-element transposition. AB - The P transposable element of Drosophila melanogaster has a complex array of cis acting DNA sequences necessary for efficient transposition. At the 3' end these sequences extend over more than 150 bp and include 11- and 31-bp sequences found repeated in inverted orientation at the 5' end. The P element's 5' end, however, cannot function as its 3' end. When two 3' P-element ends are present, the more proximal end is used preferentially. We found also that the duplication of the target site does not appear to play a role in forward transposition. PMID- 2545525 TI - DNA-binding activity of the adenovirus-induced E4F transcription factor is regulated by phosphorylation. AB - Previous experiments have identified E4F, an inducible cellular factor that binds to sequences in the adenovirus E4 promoter that are critical for E1A-dependent transcriptional activation. The E4F factor has been purified and shown to stimulate transcription in vitro from the E4 promoter. Analysis of the affinity purified factor identifies a single polypeptide of 50 kD that has E4F-specific binding activity. E4F binding activity is also regulated during F9 cell differentiation and can be activated in differentiated F9 cells by viral infection. Furthermore, the activation process appears to involve a phosphorylation event, because treatment of E4F with alkaline phosphatase abolishes activity and incubation of the phosphatase-inactivated factor with an extract from virus infected cells restores activity. PMID- 2545528 TI - A trans-acting factor required for cAMP-induced gene expression in Dictyostelium is regulated developmentally and induced by cAMP. AB - We have identified a nuclear activity that binds specifically to a GT-rich sequence or G-box shown previously by use of deletion analysis to be required for cAMP and for developmentally induced expression of the prestalk gene pst cathepsin (CP2). We show that the insertion of an oligonucleotide that contains the CP2 G-box restores regulated expression whereas the insertion of oligonucleotides that contain mutations in some of the G residues does not. Moreover, the mutant oligonucleotides do not compete for binding of the factor to the wild-type sequence. The activity of the G-box binding factor (GBF) is regulated developmentally with induction of activity occurring at the time of induction of pst-cathepsin expression. In a single-cell culture, GBF activity is inducible by cAMP, and its appearance is inhibited by cycloheximide, which suggests that the factor, or a protein component required for binding of the factor, is directly induced by cAMP and may be the rate-limiting factor required for cAMP induction of pst-cathepsin expression. Models for cAMP induction of prestalk genes are described. PMID- 2545529 TI - [Hygienic aspects of the use of sludge and liquid manure in agriculture]. PMID- 2545530 TI - [Epidemiologic significance of sludge for the transmission of zoonoses]. AB - Sludge contains all types of pathogenic organisms originating from the drainage system of the population. Decontamination is difficult and expensive, it is ranging from thermal-chemical to radioactive treatment of the materials. Suitable treatment and use of sludge can really reduce the health hazards to the population. PMID- 2545531 TI - Cloning and expression in Escherichia coli of an esterase-coding gene from the oil-degrading bacterium Acinetobacter calcoaceticus RAG-1. AB - A putative esterase gene (est) from Acinetobacter calcoaceticus RAG-1 has been cloned into Escherichia coli. Esterase-positive clones exhibited high levels of esterase activity even in intact cells. In addition, expression of the est gene conferred on E. coli the ability to grow on simple triglycerides such as triacetin (TAC). The original esterase-positive plasmid pRA17 carried a 2.2-kb insert from a partial MboI digest of RAG-1 DNA, which gave a single band with RAG 1 DNA following Southern hybridization. By subcloning and sequencing the est gene was found to contain a sequence of 870 bp which could be translated to yield a protein of Mr 32,700. In support of the sequencing results was the finding that when pRA17 was expressed in minicells, a unique peptide of Mr 32,500 was identified. This peptide was not found in minicells transformed with esterase negative plasmids, such as pRA176, which contained a Tn5 insertion in the est gene. The fact that the production of active esterase depended on the orientation of the est gene within the vector suggested that transcription proceeded from the tet promoter in pBR322. PMID- 2545532 TI - Construction of an EBNA-producing line of well-differentiated human hepatoma cells and of appropriate Epstein-Barr virus-based shuttle vectors. AB - Using cloned Epstein-Barr nuclear antigen 1 (EBNA) and oriP elements from the Epstein-Barr virus (EBV) in conjunction with liver-specific growth media, we have constructed an EBNA-producing line of well-differentiated human hepatoma cells (Hep-EBNA-2) and appropriate EBV-oriP vectors. These vectors, pBEDC1 and pBEUG1, were maintained as free extrachromosomal elements only in cells that expressed the trans-acting EBNA protein. They were readily rescued from transfected Hep EBNA-2 cells upon transformation of recA- Escherichia coli with cellular low-Mr DNA. They are true shuttle vectors in that they can propagate as free closed circular elements in both human Hep-EBNA-2 cells and E. coli. Finally, we have demonstrated the vector capability of our shuttle system by inserting into the SV40 expression cassette of pBEUG1 a large full-length cDNA encoding coagulation factor VIII. Our data clearly show that EBV-oriP episomes are able to stably propagate in an hepatic background and that neither high levels of EBNA protein nor multiple copy episomes significantly interfere with the expression of the set of hepatic functions that have been analyzed. These results are discussed in terms of gene amplification and cloning of genes that program liver differentiation. PMID- 2545533 TI - The construction of recombinant industrial yeasts free of bacterial sequences by directed gene replacement into a nonessential region of the genome. AB - The yeast SMR1 gene was used as a dominant resistance-selectable marker for industrial yeast transformation and for targeting integration of an economically important gene at the homologous ILV2 locus. A MEL1 gene, which codes for alpha galactosidase, was inserted into a dispensable upstream region of SMR1 in vitro; different treatments of the plasmid (pWX813) prior to transformation resulted in 3' end, 5' end and replacement integrations that exhibited distinct integrant structures. One-step replacement within a nonessential region of the host genome generated a stable integration of MEL1 devoid of bacterial plasmid DNA. Using this method, we have constructed several alpha-galactosidase positive industrial Saccharomyces strains. Our study provides a general method for stable gene transfer in most industrial Saccharomyces yeasts, including those used in the baking, brewing (ale and lager), distilling, wine and sake industries, with solely nucleotide sequences of interest. The absence of bacterial DNA in the integrant structure facilitates the commercial application of recombinant DNA technology in the food and beverage industry. PMID- 2545534 TI - Production of salivary type alpha-amylase in human lung cancer. AB - alpha-Amylase, which is produced by lung cancer tissue, was studied by cloning cDNAs from a cell line originating from lung cancer that produces amylase. Sequencing studies with this cDNA showed that the expressing gene is of the salivary type. The specific location of the start point of transcription, as revealed by S1 mapping, supported this conclusion. PMID- 2545536 TI - A sensitive and simple assay for neomycin phosphotransferase II activity in transgenic tissue. AB - A new assay for the detection of the enzyme neomycin phosphotransferase II (NPTII) in crude cell extracts is described. The method is based on the chromatographic separation of the compounds resulting from the reaction of NPTII with kanamycin (Km) and [gamma-32P]ATP; the labelled Km.phosphate is subsequently detected by autoradiography. Chromatography is carried out on polyethyleneimine cellulose plates. This assay has been tested with bacterial, plant and animal crude extracts and comparisons with the assays in current use have been made. Our assay has several advantages: it is simple, rapid, sensitive and safe. PMID- 2545535 TI - Cooperative interactions between the GRP78 enhancer and promoter elements in hamster fibroblasts. AB - A non-tissue-specific enhancer derived from the promoter of the rat 78-kDa glucose-regulated protein (GRP78)-coding gene was tested for its ability to stimulate the activity of its homologous promoter and two heterologous promoters (simian virus 40 and mouse mammary tumor virus). Single and double copies of the enhancer were inserted at positions 5' and 3' of the cat-expression vectors under the direction of the above promoters. The recombinant plasmids were transfected into hamster fibroblast K12 cells and assayed for chloramphenicol acetyl transferase activity under induced and non-induced conditions. We report that the GRP78 enhancer (i) exhibits strong cooperative interactions with its homologous promoter; (ii) can activate and confer a calcium ionophore (A23187) inducibility to heterologous promoters in an orientation-independent manner; (iii) prefers the 5' over the 3' location and; (iv) is dosage dependent in that two copies are twice as active as a single unit. PMID- 2545537 TI - Cloning and characterization of the Escherichia coli phosphoglycerate kinase (pgk) gene. AB - The pgk gene of Escherichia coli coding for the phosphoglycerate kinase was subcloned from the Carbon and Clarke collection plasmid pLC33-5. The position and direction of transcription of the pgk gene was determined by Tn5 insertion mutagenesis. Analysis of proteins encoded from these plasmids showed that the pgk gene product is a 40-kDa protein, and that the gene is transcribed from two promoters, one immediately in front of the gene and one in front of an upstream gene coding for a 38-kDa polypeptide of unknown function. The position of the Pgk protein on two-dimensional O'Farrel gels was identified, and from this we conclude that it is one of the proteins induced by anaerobiosis [Smith and Neidhardt, J. Bacteriol. 154 (1987) 336-343]. The pgk gene was also found to show growth phase regulation; the synthesis of Pgk protein was induced more than ten fold during transition from the exponential to the stationary growth phase. PMID- 2545538 TI - The C-terminal part of a gene partially homologous to CDC 25 gene suppresses the cdc25-5 mutation in Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, the product of the CDC25 gene is required for progression in the cell division cycle. It is necessary for cAMP production. It has been suggested that the CDC25 gene product acts through Ras proteins. We report the cloning of a DNA fragment from a new gene able to suppress the thermosensitive phenotype of the cdc25-5 mutation. It is unable to suppress the defect of a mutant of the adenylate cyclase gene or of the ras1, ras2ts double mutant. This DNA fragment prevents the drop in cAMP level in cdc25-5 mutant cells shifted to restrictive temperature. The complementing part of this fragment contains a truncated open reading frame (ORF) corresponding to the 3' end of a gene we named SCD25. The 584-amino acid sequence deduced from this ORF shares 45% identity with the 592-aa C-terminal part of the CDC25 ORF which is sufficient for complementation of cdc25 mutations. Some of the common sequences between these two genes are also partially homologous with the amino acid sequence of LTE1, another gene of S. cerevisiae. The capacity of the SCD25 fragment to suppress a cdc25 mutation and its homology to the C-terminal part of the CDC25 led us to propose that the CDC25 and the SCD25 C-terminal fragments each encode a protein domain which is capable in itself to support a similar biochemical function. PMID- 2545539 TI - Helpers for efficient encapsidation of SV40 pseudovirions. AB - Plasmid DNA that carries the simian virus 40 (SV40) ori can be packaged as SV40 pseudovirions. The pseudovirions are very efficient in gene transmission into a variety of cell types, including human hemopoietic cells. They are routinely prepared with wild-type (wt) SV40 as a helper. In the present study, several parameters required for the helper function were investigated. Plasmids that carry pBR322 sequences in addition to the late genes of SV40 were inefficient in providing helper functions, presumably because the prokaryotic sequences interfered with expression of the SV40 late genes. Efficient helpers were plasmid pSVPiC [Villarreal and Soo, Mol. Appl. Genet. 3 (1985) 62-71] and an SV40 defective virus SLT3 (presently constructed). Plasmid pSVPiC carries a duplication of the SV40 ori and enhancer regions, and pi AN7 sequences. Because of its large size it was not packaged into virion particles. However, it underwent extensive recombination generating infective SV40 particles. Almost no prokaryotic sequences are included in SLT3, that carries the SV40 late gene. In spite of its small size (3.5 kb) it was packaged efficiently, creating defective (T-antigen-negative) SV40 virions. The availability of T-antigen positive and negative pseudovirion mixtures enabled us to suggest that T-antigen drives gene amplification in the target human hemopoietic cells. PMID- 2545541 TI - [Crystalline and amorphous forms of SiO2 in the lungs of patients with silicosis]. AB - Lung samples of dead patients with silicosis (9 persons) and those without silicosis were compared. The obtained results showed that there was no significant difference in the content of crystalline and amorphous forms of SiO2 in the control group and the dust-exposed group. Because of elevated content of SiO2 amorphous form total SiO2 concentrations were higher in the lungs of patients with silicosis than in the control group, but the content of the crystalline form didn't differ significantly. The samples were analyzed on the basis of the technique used with slight modifications for the determination of SiO2 crystalline form in industrial dust. Before the analysis was carried out the conditions for primary sample treatment were standardized. It was also shown that the mass of ashed lungs constituted 0.8 +/- 0.2% of the damp mass and 2.4 +/- 0.7% of the dry one. PMID- 2545540 TI - [Problems of improving working conditions for workers engaged in the production of nitrogen mineral fertilizers]. AB - The study results show that significant improvement of labour conditions can be achieved due to the introduction of a new distribution pattern of technological equipment, further improvement of equipment supply of the technological process, further mechanization of production operations, rational arrangement and disposition of control desks for technological processes, utilization of possible means of protection against noise, improvement of measuring machines, rational organization of the workplace for packing of stock-produced products and application of the means of small-scale mechanization, rational distribution of the sections of packing, filling and loading. PMID- 2545542 TI - [Electrophysical processes during grinding of materials possessing piezoelectric properties, and fibrogenic activity of the dust (review of the literature)]. PMID- 2545543 TI - [A crystallographic method of examination in diagnosing the bronchospastic component of allergic origin of chronic dust-induced bronchitis]. PMID- 2545544 TI - [Status of the upper respiratory tract and skin of workers manufacturing artificial mineral wool]. PMID- 2545545 TI - [Problem of correlating the effect of carbon fiber dust on the body and the degree of migration of residual monomers and the products of their thermal destruction in a model biological media]. PMID- 2545546 TI - [Clinico-electroneuromyographic characteristics of occupational polyneuropathies of toxico-allergic origin]. PMID- 2545547 TI - [Comparative carcinogenic properties of basalt fiber and chrysotile-asbestos]. AB - In order to eliminate asbestos adverse effect on workers' health it was necessary to use mineral rayon, primarily basalt fibre, instead of asbestos. During a chronic experiment on animals the oncogenicity of 2 kinds of basalt fibre was studied compared to chrysotile asbestos. The dust dose of 25 mg was twice administered by intraperitonial route. All types of dust induced the onset of intraperitonial mesotheliomas but neoplasm rates were significantly lower in the groups exposed to basalt fibre. There was no credible data on the differences between the groups exposed to various types of basalt fibre. Since the latter produced some oncogenic effect, it was necessary to develop a complex of antidust measures, fully corresponding to the measures adopted for carcinogenic dusts. PMID- 2545548 TI - ESR studies of structure and kinetics of radicals from hydroxyurea. An antitumor drug directed against ribonucleotide reductase. AB - Hydroxyurea (HU) is a clinically applied antineoplastic drug, which quenches tyrosine radicals in the active site of ribonucleotide reductase (RR) and inhibits DNA synthesis in proliferating cells. Under oxidizing conditions (Cu2+ or H2O2) long-lived radicals from HU have been found by ESR. The structure of HU radicals was established to be: (formula; see text). The kinetics of formation and decay of HU radicals after reaction of HU with H2O2 is complex; it exhibits a lag-phase, a maximum, and a decay, all depending on the concentration of HU. Biological consequences of HU radicals for the inhibition of RR as well as their role in cytotoxic events during chemotherapy of cancer are discussed. PMID- 2545549 TI - Detection of oxygen radicals during reperfusion of intestinal cells in vitro. AB - The nitroxide OXANO. (2-Ethyl-2,5,5-trimethyl-3-oxazolidinoxyl) which in its reduced form, OXANOH (2-Ethyl-1-hydroxy-2,5,5-trimethyl-3-oxazolidine), is capable of reacting with short-lived radicals, forming a secondary stable radical, was used for ESR-detection of radical production in isolated cells. The properties of OXANO. and OXANOH in terms of stability in cellular and subcellular systems, membrane permeability and effects on cellular viability were evaluated. Ischemia and reperfusion was simulated in vitro in a preparation of cells from rat intestinal mucosa by incubation at high density (4 X 10(8) cells/ml) under an atmosphere of nitrogen for 25 min and resuspended with fresh oxygenated buffer containing 5 mM OXANOH. A significant increase in radical formation during the 15 min reperfusion period studied was obtained in cells exposed to ischemia compared to control cells incubated at normal density under an atmosphere of oxygen. The addition of 5 microM of the scavenging enzyme superoxide dismutase reduced the radical formation by 50%. The time sequence of the superoxide formation was calculated as the difference in radical production in the presence and absence of superoxide dismutase. PMID- 2545550 TI - Intermediates in the aerobic autoxidation of 6-hydroxydopamine: relative importance under different reaction conditions. AB - Autoxidation of 6-hydroxydopamine (6-OHDA) proceeds through a balanced network of: transition metal ions, superoxide, hydrogen peroxide, hydroxyl radicals, and other species. The contribution of each to the reaction mechanism varies dramatically depending upon which scavengers are present. The contribution of each propagating intermediate increases when the involvement of others is diminished. Thus, superoxide (which is relatively unimportant when metal ions can participate) dominates the reaction when transition metal ions are bound (especially at higher pH), and it becomes essential in the simultaneous presence of catalase plus chelators. Transition metal ions participate more if superoxide is excluded; hydrogen peroxide becomes more important if both .O2- and metal ions are excluded; and hydroxyl radicals contribute more to the reaction mechanism if both H2O2 and .O2- are excluded. Superoxide dismutase inhibited strongly, by two distinct mechanisms: a high affinity mechanism (less than 13% inhibition) at catalytically effective concentrations, and a low affinity mechanism (almost complete inhibition at the highest concentrations) which depends upon both metal binding and catalytic actions. In the presence of DETAPAC catalytic concentrations of superoxide dismutase inhibited by over 98%. Conversely, metal chelating agents inhibited strongly in the presence of superoxide dismutase. When present alone they stimulated (like EDTA), inhibited (like desferrioxamine), or had little effect (like DETAPAC). Catalase which stimulated slightly but consistently (less than 5%) when added alone, inhibited 100% in the presence of superoxide dismutase + DETAPAC. However, in the absence of DETAPAC, catalase decreased inhibition by superoxide dismutase, yielding a 100% increase in reaction rate. Hydroxyl scavengers (formate, mannitol or glucose) alone produced little or no (less than 10%) inhibition, but inhibited by 30% in the presence of catalase + superoxide dismutase. Paradoxically, they stimulated the reaction in the presence of catalase + superoxide dismutase + DETAPAC. PMID- 2545551 TI - Interaction of lactoperoxidase with hydrogen peroxide. Formation of enzyme intermediates and generation of free radicals. AB - Peroxidases belong to a group of enzymes which catalyze the oxidation of numerous organic and inorganic substrates by hydrogen peroxide. Most peroxidases, including lactoperoxidase (LPO), contain ferriprotoporphyrin IX as a prosthetic group. A characteristic feature of hemoprotein peroxidases is their ability to exist in various oxidation states. There are five known enzyme intermediates. In increasing order of their oxidative equivalents these are ferrous enzyme, ferric or native enzyme, Compound II, Compound I, and Compound III (sections 5, 7). They are readily distinguished from each other by their absorbance in the Soret region (380-450 nm) and visible range (450-650 nm). In the course of Compound III and Compound II conversion back to the native peroxidase, oxygen derived free radicals such as O2-, HO.2, and .OH are generated. Simultaneously the enzyme is irreversibly damaged. In the presence of an exogenous electron donor, such as iodide, the interconversion between the various oxidation states of the peroxidase is markedly affected. Compound II and/or Compound III formation is inhibited, depending on the H2O2 concentration. In addition, the enzyme is largely protected from irreversible inactivation. These effects of iodide are readily explained by 1) the two-electron oxidation of iodide to Iox by Compound I, which bypasses Compound II as an intermediate, and 2) the rapid oxidation of H2O2 to O2 by the oxidized species of iodide which prevents the generation of oxygen derived free radicals. PMID- 2545552 TI - Dimethylthiourea prevents hydrogen peroxide and neutrophil mediated damage to lung endothelial cells in vitro and disappears in the process. AB - Dimethylthiourea (DMTU) progressively disappeared following reaction with increasing amounts of hydrogen peroxide (H2O2) in vitro. DMTU disappearance following reaction with H2O2 was inhibited by addition of catalase, but not aminotriazole-inactivated catalase (AMT-catalase), superoxide dismutase (SOD), mannitol, benzoate or dimethyl sulfoxide (DMSO) in vitro. By comparison, DMTU disappearance did not occur following addition of histamine, oleic acid, elastase, trypsin or leukotrienes in vitro. Addition of DMTU also decreased H2O2 mediated injury to bovine pulmonary artery endothelial cells (as reflected by LDH release) and DMTU disappeared according to both added amounts of H2O2 and corresponding degrees of injury. DMTU disappearance was also relatively specific for reaction with H2O2 in suspensions of endothelial cells where it was prevented by addition of catalase, but not AMT-catalase or SOD and did not occur following sonication or treatment with elastase, trypsin or leukotrienes. Addition of washed human erythrocytes (RBC) also prevented both H2O2 mediated injury and corresponding DMTU decreases in suspensions of endothelial cells. In addition, phorbol myristate acetate (PMA) and normal neutrophils, but not O2 metabolite deficient neutrophils from patients with chronic granulomatous disease (CGD), caused DMTU disappearance in vitro which was decreased by simultaneous addition of catalase, but not SOD, sodium benzoate or DMSO. Finally, addition of normal neutrophils (but not CGD neutrophils) and PMA caused DMTU disappearance and increased the concentrations of the stable prostacyclin derivative (PGF1 alpha) in supernatants of endothelial cell suspensions. In parallel, DMTU also decreased PMA and neutrophil-mediated PGF1 alpha increases in supernatants from endothelial cell monolayers. Our results indicate that DMTU can decrease H2O2 or neutrophil mediated injury to endothelial cells and that simultaneous measurement of DMTU disappearance can be used to improve assessment of the presence and toxicity of H2O2 as well as the H2O2 inactivating ability of scavengers, such as RBC, in biological systems. PMID- 2545553 TI - Adhering lung macrophages produce superoxide demonstrated with desferal-Mn(IV). AB - It has been shown that H2O2, the dismutation product of O2., is produced at cell surface interfaces. Nevertheless, the relationships between the degree of attachment itself, type of surface, and O2. production are not clear. Superoxide production can be measured by the O2.-dependent reduction of nitroblue tetrazolium to an insoluble formazan. Superoxide dismutase (SOD) may be unable to scavenge O2. produced between alveolar macrophages (AM) and a surface. Desferal Mn(IV) (Des-Mn), a low molecular weight mimic of SOD, is protective against paraquat toxicity in vivo, presumably because of specificity for O2-. Using that assumption, Des-Mn was used to measure O2. production that occurred during adherence of AM. AM suspensions were placed on fibronectin-coated glass coverslips or uncoated glass coverslips or non-stick tissue culture plates. Adherence to the surfaces varied with fibronectin greater than glass greater than non-stick and the percent formazan positive cells was 60, 24, and 4, respectively. With SOD present, the percentage of formazan positive cells were 40, 17, and 2; however, in the presence of Des-Mn the percent stained cells was 4, 4, and 0. When phorbol myristate acetate (PMA) was added during adherence, the percent of formazan positive cells was 82, 57, and 44, respectively. With PMA, Des-Mn was able to inhibit 88-100% of formazan staining whereas SOD inhibition decreased more markedly with increasing adherence. These results indicated that the degree of attachment correlated with both the degree of NBT reduction and the relative effectiveness of Des-Mn versus SOD to scavenge O2.. PMID- 2545555 TI - Structure, acid/base properties and transformation reactions of purine radicals. PMID- 2545554 TI - The SO4- -induced oxidation of 1,3,6-trimethyluracil and 1,3-dimethylthymine by potassium peroxodisulphate and oxygen in aqueous solution: an interesting contrast. PMID- 2545556 TI - Theoretical studies of molecular mechanisms of DNA damage induced by hydroxyl radicals. PMID- 2545557 TI - Radiation-induced release of undamaged uracil from polyuridylic acid and the formation of phosphomonoester end groups in irradiated polynucleotides. PMID- 2545558 TI - ESR studies on the mechanism of .OH-induced strand breakage of poly(U). PMID- 2545559 TI - Rate and yield of OH-induced strand break formation of polynucleotides and DNA. PMID- 2545560 TI - DNA damage in frozen aqueous solution: sequence dependence and end groups. PMID- 2545561 TI - Hydroxyl radical-induced crosslinking between double-stranded poly(dA-dT) and tripeptides containing an aromatic residue. PMID- 2545562 TI - Radical-induced DNA damage and specific implications in evaluating genetic changes. PMID- 2545563 TI - Radiation chemistry of adenine derivatives following direct ionization in solids: ESR and ENDOR investigations. PMID- 2545564 TI - ESR and ENDOR studies of X-irradiated single crystals of guanine derivatives. PMID- 2545565 TI - Free radical formation in single crystals of 2'-deoxyguanosine 5'-monophosphate, and guanine hydrobromide monohydrate after X-irradiation at 10 and 65 K: an ESR, ENDOR and FSE study. PMID- 2545567 TI - Free radicals formed by electron gain in oligomers of DNA. PMID- 2545566 TI - Free radical formation in nucleosides and nucleotides of guanine: ESR and ENDOR of guanosine 5'-monophosphate and guanosine: dimethylformamide X-irradiated at 10 K. PMID- 2545568 TI - Matrix-isolation of free radicals from purines and derivatives in aqueous low temperature glasses. PMID- 2545569 TI - Spin trapping of sugar radicals in solid and aqueous phases. PMID- 2545570 TI - ESR investigations of the reactions of radiation-produced thiyl and DNA peroxyl radicals: formation of sulfoxyl radicals. PMID- 2545571 TI - [International symposium: Fluconazole in fungal infections. 20-21 January 1989, Berlin. Abstracts]. PMID- 2545572 TI - Changes in angiotensin-converting enzyme during pregnancy in the guinea pig. AB - Angiotensin-converting enzyme (ACE) activity was measured weekly in plasma of pregnant guinea pigs either during the last half of pregnancy (experiment 1) or throughout pregnancy (experiment 2). ACE activity increased about the time of the first missed estrus, reached a maximum about days 35-40, and then declined. A similar rise and decline in plasma ACE activity does not appear to have been reported in other animals or in humans. PMID- 2545573 TI - Variability of cortisol and adrenocorticotropic hormone responses to metoclopramide during the menstrual cycle. AB - Responses of cortisol and adrenocorticotropic hormone (ACTH) to metoclopramide (MCP) were examined in normal women throughout the menstrual cycle. The administration of MCP increased circulating levels of cortisol and ACTH in normal women during the mid-luteal but not during the early and late follicular phases. MCP may stimulate ACTH release through an antidopaminergic mechanism, and endogenous dopaminergic tone is likely to be increased during the luteal phase compared to the follicular phase. Consequently, the positive responses of cortisol and ACTH to MCP observed in the subjects during the mid-luteal phase may be related to the increased dopaminergic tone. PMID- 2545574 TI - Relevance of ultrastructural immunocytochemistry in the characterization of unclassifiable leukemias: correlation with phenotypic and genic studies. AB - Nine cases of acute leukemia presenting unusual phenotype were studied by light microscopy (LM) cytochemistry and transmission electron microscopy (TEM) immunocytochemistry with the immunogold staining (IGS) method; in addition, cytogenetic and molecular analyses were performed. The presence of myeloperoxidase (MPO) was studied at TEM in combination with immunophenotype to identify minor populations not characterizable at LM. Four of nine cases had no TEM/MPO reactivity, whereas the remaining five showed variable percentages of positive cells. Of the MPO negative cases, one was a megakaryoblastic leukemia with a positive platelet peroxidase (PPO) reaction, and three were lymphoid. Among the peroxidase positive cases, the percentage of MPO reactive cells was higher at TEM than at LM examination. In case 5 TEM analysis indicated that cells with some MPO reactivity at LM were non neoplastic myeloid cells. With this combined technique in cases 1 and 2 we excluded the presence of the MPO enzyme in CD15 positive lymphoid cells and, in another case, we documented the existence of CD19/MPO positive cells. The value of cytochemistry and immunology at the ultrastructural level for the characterization of blast cells and for the precise diagnosis of leukemia with "unusual" phenotype is illustrated. PMID- 2545575 TI - Reactive hemophagocytosis in Ki-l positive large cell lymphoma: a case study. AB - A case of large cell lymphoma presenting with hemophagocytic syndrome is reported. The clinicopathological findings suggested a diagnosis of malignant histiocytosis, but on the basis of immunohistological studies Ki-l lymphoma was diagnosed. Neoplastic cells expressed activation antigens such as HLA-DR, IL 2R, T10 and Ki-l, and showed high proliferative activity, but were devoid of T and B cell markers. The high percentage of reactive macrophages found in the bone marrow and lymph node probably reflected the release of lymphokines by the tumor population. The patient was treated with aggressive chemotherapy and is in complete remission at 8 months from diagnosis. PMID- 2545576 TI - Aplastic crisis caused by B19 virus in a child during induction therapy for acute lymphoblastic leukemia. AB - A child undergoing induction therapy for acute lymphoblastic leukemia suffered an aplastic crisis associated with B19 virus infection. Good response to the antiblastic therapy led to a burst in erythropoiesis favoring high viral replication, which was responsible for strong erythroblastic inhibition and severe viremia. The patient's B19 antibody response became evident very late, probably because of the antiblastic effect of the therapy; nevertheless, recovery was complete in little more than a week, favored by B19 IgG transfusion with a red blood cell concentrate. This report suggests that immunosuppressed subjects, as well as those suffering from hemolytic anemia must also be considered "at risk" for aplastic crisis due to B19 infection. PMID- 2545577 TI - A sensitive in vivo platelet function test in rats based on intravenously injected collagenase. AB - The transient decrease of the platelet concentration in the flowing blood after intravenous injection of collagenase was used as a measure for the effectiveness of the platelet-vessel wall interactions in rats. The decrease of the concentration in circulating platelets was regarded as an expression of the in vivo platelet function. The influence of acetylsalicylic acid, imidazole, indomethacin, ketanserin, RA 233, and dipyridamole on this test was investigated in anaesthetized rats. All tested drugs showed an effect on the extent of the platelet concentration decrease after the collagenase injection compared to the control values, but no drug was significantly effective. This test could prove to be a valuable tool for testing the efficiency of antithrombotic drugs in vivo. PMID- 2545578 TI - [Inhibitory effect of N-alpha-[(S)-1-carboxy-3-phenylpropyl]-L-lysyl-L-proline (MK-0521) on angiotensin converting enzyme in vitro]. AB - N-alpha-[(S)-1-carboxy-3-phenylpropyl]-L-lysyl-L-proline (MK-0521) is a new non sulfhydryl-containing angiotensin converting enzyme (ACE) inhibitor. The present investigation describes its ACE and other enzymes inhibitory properties and compares it to those of captopril, MK-421 and MK-422 in vitro. MK-0521 inhibited rat pulmonary ACE by 50% (IC50) at a concentration of 3 nM and was 6.13 times more potent than captopril. The IC50 values of MK-421 and MK-422 against ACE were 2,000 nM and 3.5 nM, respectively. MK-0521 had practically no inhibitory activities against carboxypeptidase A, carboxypeptidase B, leucine aminopeptidase, papain, pepsin and trypsin. The kinetic study on the inhibitory activity of M-0521 against ACE using Lineweaver-Burk plots indicated that MK-0521 exerted competitive ACE inhibition. The dialysis study conducted on the ACE-MK 0521 complex revealed that the inhibitory effect of MK-0521 against ACE was reversible. In the guinea pig ileum, MK-0521 potentiated the contractile effect of bradykinin and depressed the contractile effect of angiotensin I. These effects on bradykinin and angiotensin I were 33.11 and 2.63 times more potent than that of captopril, respectively. The present results suggest that MK-0521 may show a potent hypotensive effect in vivo. PMID- 2545579 TI - [Angiotensin converting enzyme inhibitory activity of MK-0521 in vivo and antihypertensive effect of its single oral administration on blood pressure and effect on the renin-angiotensin system in 2-kidney Goldblatt hypertensive dogs]. AB - Angiotensin converting enzyme (ACE) inhibitory activity of MK-0521 in dogs and the effects of its single oral administration on blood pressure and the renin angiotensin system in 2-kidney Goldblatt hypertensive dogs were compared with those of captopril and MK-421. MK-0521 at 0.001-0.1 mg/kg, i.v., or 0.01-1 mg/kg, p.o., attenuated the pressor effect of angiotensin I without affecting that of angiotensin II and augmented the depressor effect of bradykinin. The potency of MK-0521 to reduce the pressor effect of angiotensin I was 9.8 times that of captopril by intravenous administration, and by oral administration, it was 15.9 32.1 times that of captopril and approximately 3 times that of MK-421. When administered orally, the onset of action and the time to peak effectiveness were more rapid than those of MK-421, but slower than those of captopril. Duration of the action of MK-0521 was longer than that of captopril and equal or longer than that of MK-421. The inhibition of ACE was well correlated with serum MK-0521 levels. MK-0521 produced a dose-dependent antihypertensive effect in 2-kidney Goldblatt hypertensive dogs at over 0.3 mg/kg, p.o., without affecting the heart rate. The antihypertensive effect of MK-0521 was persistent and approximately 3 times more potent than that of captopril. MK-0521 inhibited the serum ACE activity and increased the plasma renin activity, while it had a tendency to decrease plasma aldosterone level. These changes were parallel to the time course of the antihypertensive effect. These results suggest that the main mechanism of the antihypertensive effect of MK-0521 is the suppression of angiotensin II production due to the inhibition of the ACE. PMID- 2545580 TI - [Antihypertensive effect of repeated oral administration of MK-0521 in 2-kidney Goldblatt hypertensive dogs]. AB - MK-0521 and captopril were orally administered to acute (6 to 8 days after unilateral renal artery constriction) and chronic (2 to 2.5 months after the constriction) 2-kidney Goldblatt hypertensive dogs for 7 to 21 days. MK-0521 lowered the blood pressure to similar extents in the acute and chronic stages of hypertension. The antihypertensive effect of MK-0521 was dose-dependent and persistent even after its cessation. Captopril also produced an antihypertensive effect, although the effect in the chronic stage of hypertension was less prominent than that in the acute stage of hypertension. MK-0521 was more inhibitory on the renin-angiotensin system than captopril. In the acute stage of hypertension, the dogs treated with MK-0521 had increased angiotensin converting enzyme (ACE) activity, while they had decreased plasma angiotensin II level and elevated plasma angiotensin I level and plasma renin activity. These results clarified the inhibitory effect of MK-0521 on ACE. In contrast, in the chronic stage of hypertension, MK-0521 showed no depression of plasma angiotensin II. There were no significant changes in daily urinary volume, and renal clearances of sodium, potassium and creatinine. These results suggest that the major mechanism of the antihypertensive effect of MK-0521 in 2-kidney Goldblatt hypertensive dogs is an inhibition of the ACE. In addition, the different effects in the acute and chronic hypertensive dogs suggest that some differences exist in the mechanisms of maintaining blood pressure between the two stages of 2-kidney Goldblatt hypertensive dogs. PMID- 2545581 TI - [Pharmacological effects of reiousan on experimental hepatic injuries and hepatic functions]. AB - Pharmacological effects of Reiousan, a crude drug preparation consisting of bezoar and ginseng, on experimental hepatic injuries and hepatic functions were studied. After administration of Reiousan, carbon tetrachloride, d-galactosamine and alpha-naphthylisothiocyanate-induced experimental hepatic injuries in rats were inhibited. Facilitation of recovery from increased retention rate of sulfobromophthalein in hepatectomized rats, increase in hepatic blood flow, inhibition of superoxide anions, and decrease in blood ethanol concentration in rats administered ethanol were observed after application of Reiousan. Inhibitory effects of Reiousan on experimental hepatic injuries may result from the increase in hepatic blood flow and inhibition of superoxide anions. PMID- 2545582 TI - [Neurochemical studies on bifemelane, a new cerebral function improver. I. Effects of bifemelane on the function of neurotransmission-related enzymes and receptors in rat brain]. AB - We examined neurochemically the effects of bifemelane (BF) on muscarinic ACh (mACh-R) and beta-adrenergic receptors (beta-AdR) and imipramine binding sites and the activities of acetylcholinesterase (AChE), choline acetyltransferase (CAT) and monoamine oxidase (MAO) in the P2 fractions of rat brain, ex vivo and in vitro. Male rats were given daily injections of 10, 30 mg/kg BF, p.o., for a period of 4 weeks. The Kd and Bmax values for mACh-R in the rat forebrain by administration of 10, 30 mg/kg BF decreased significantly compared with that of the control, although the Kd and Bmax values for beta-AdR and imipramine binding sites were almost identical. The Km and Vmax values of A- and B-form MAO decreased in rats that had been administered 30 mg/kg BF for 4 weeks. The binding of 3H-QNB (quinuclidinyl benzilate) on mACh-R, 125I-CYP (iodocyanopindolol) on beta-AdR and 3H-imipramine on imipramine binding sites decreased by 60, 20 and 70% in the presence of 1 microM BF, respectively, while the addition of 1 microM BF inhibited MAO activity by about 50%. However, CAT and AChE activities were not inhibited by BF. PMID- 2545583 TI - Proteins of the acrosomal region in mouse sperm: immunological probes reveal post testicular modifications. AB - Due to the central role the acrosomal region plays in sperm-egg interactions, monoclonal antibodies (mAbs) were used to identify components of this domain in mouse sperm. Several sperm proteins that localize specifically to the anterior acrosomal region are described here in terms of electrophoretic mobility, susceptibility to proteolytic degradation, and post-translational modification during epididymal transit. Six different mAbs were used, each recognizing a distinctive antigen (Ag) or set of Ags in cauda epididymal mouse sperm: a doublet of 185/200 Kd (M42 mAb); 150-160 Kd (M5 mAb); 105 Kd (W71 mAb); 21, 35, and 60 Kd (M41 mAb); 27 and 33 Kd (W33 mAb); and 57 and 86 Kd (W108 mAb). Previously reported work implicates two of these, M42 Ag and M5 Ag, as participants in sperm zona interaction (Saling and Lakoski: Biol Reprod 33:527-536, 1985; Saling: Dev Biol 117:511-519, 1986; and Lakoski et al.: Biol Reprod 38:221-233, 1988). Recognition of some (M42, M5, W108), but not all (W33), of the Ags by their corresponding mAbs was affected by sperm incubation with proteases (trypsin or collagenase). Evidence of post-translational modification during epididymal maturation was suggested by altered electrophoretic mobility of several of the Ags (M42, M5, W33, and W108) accompanying sperm transit from proximal to distal epididymis. Retention of sperm within the caput epididymis prevented structural alterations for the four proteins examined, indicating that spatial rather than temporal factors are critical for Ag modification in maturing mouse sperm. PMID- 2545584 TI - Role of glutathione peroxidase in protecting mammalian spermatozoa from loss of motility caused by spontaneous lipid peroxidation. AB - Mouse and human spermatozoa, but not rabbit spermatozoa, have long been known to be sensitive to loss of motility induced by exogenous H2O2. Recent work has shown that loss of sperm motility in these species correlates with the extent of spontaneous lipid peroxidation. In this study, the effect of H2O2 on this reaction in sperm of the three species was investigated. The rate of spontaneous lipid peroxidation in mouse and human sperm is markedly enhanced in the presence of 1-5 mM H2O2, while the rate in rabbit sperm is unaffected by H2O2. The enhancement of lipid peroxidation, the rate of reaction of H2O2 with the cells, the activity of sperm glutathione peroxidase, and the endogenous glutathione content are highest in mouse sperm, intermediate in human sperm, and very low in rabbit sperm. Inactivation of glutathione peroxidase occurs in the presence of H2O2 due to complete conversion of endogenous glutathione to GSSG: No GSH is available as electron donor substrate to the peroxidase. Inactivation of glutathione peroxidase by the inhibitor mercaptosuccinate has the same effect on rate of lipid peroxidation and loss of motility in mouse and human sperm as does H2O2. This implies that H2O2 by itself at 1-5 mM is not intrinsically toxic to the cells. With merceptosuccinate, the endogenous glutathione is present as GSH in mouse and human sperm, indicating that the redox state of intracellular glutathione by itself plays little role in protecting the cell against spontaneous lipid peroxidation. Mouse and human sperm also have high rates of superoxide production. We conclude that the key intermediate in spontaneous lipid peroxidation is lipid hydroperoxide generated by a chain reaction initiated by and utilizing superoxide. Removal of this hydroperoxide by glutathione peroxidase protects these sperm against peroxidation; inactivation of the peroxidase allows lipid hydroperoxide to increase and so increases the peroxidation rate. Rabbit sperm have low rates of superoxide reaction due to high activity of their superoxide dismutase; lack of endogenous glutathione and low peroxidase activity does not affect their rate of lipid peroxidation. As a result, these sperm are not affected by either H2O2 or mercaptosuccinate. These results lead us to postulate a mechanism for spontaneous lipid peroxidation in mammalian sperm which involves reaction of lipid hydroperoxide and O2 as the rate-determining step. PMID- 2545585 TI - Enhancement of assays of activities of endonucleases on defined substrates by Poisson and non-Poisson combinatoric analysis. AB - Assay of endonuclease activity, as performed in most laboratories, depends upon change in form of small, defined substrate molecules, with a secondary computation required to obtain a determination of enzyme activity. We now explore the assumptions inherent in these computations and provide a series of equations that permit more accurate determinations of enzyme activity from assays of this type. These equations allow information to be obtained not only from substrate fractions left uncleaved by the endonuclease, upon which conventional systems rely, but also from products cleaved by the enzyme. Some information yielded by these equations is unobtainable using conventional methods of analysis. PMID- 2545586 TI - Up-regulation of IGF-I receptors on IM-9 cells by IGF-II peptides. AB - To examine a possible role for IGF-II in the regulation of IGF-I receptors we measured 125I-IGF-I binding on IM-9 cells following pre-incubation with IGF II/IGF-I mixtures, purified MSA (a rat IGF-II-like peptide), pure IGF-I, or insulin. Whereas all preparations tested induced down-regulation of IGF-I binding after 20 hours, distinct differences were noted after six hour pre-incubation: IGF-I (100 ng/ml) and insulin (1 microgram/ml) both induced down-regulation of IGF-I binding (15 +/- 2% and 19 +/- 2% respectively). However, a mixture of IGF II and IGF-I (100 ng/ml each) induced consistent up-regulation of IGF-I binding (16 +/- 2%) (mean +/- SE, n = 14), and a preparation enriched in IGF-II (250 ng/ml IGF-II and 75 ng/ml IGF-I) induced 20 +/- 5% (n = 3) up-regulation at six hours. Purified MSA (200 ng/ml) induced 15% up-regulation of IGF-I binding at six hours. Scatchard analysis of displacement curves showed that increased binding was due to loss of low affinity binding, with enhancement of high affinity sites. The up-regulation of IGF-I binding was unaffected by treatment with 0.1 mM cycloheximide, but was blunted by 5 microM colchicine. It is concluded that 1. IGF-II induces up-regulation of IGF-I receptors on IM-9 cells following 6 hour pre-incubation; 2. This phenomenon is not mimicked by the structurally-related peptides IGF-I or insulin; The up-regulation is due to enhanced high affinity binding sites.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545587 TI - Autoradiographic analysis of GABA-benzodiazepine receptors in an animal model of acute hepatic encephalopathy. AB - To complement analogous studies using conventional ligand-membrane binding assays, the densities of gamma-aminobutyric acid and benzodiazepine receptors in the brain have been assessed using an autoradiographic technique in an animal model of hepatic encephalopathy. Hepatic encephalopathy due to fulminant hepatic failure was induced in rabbits by the intravenous injection of galactosamine. The specific binding of three radiolabeled ligands was assessed densitometrically in several microregions of cerebral cortex, hippocampus and cerebellum. [3H]Muscimol was used to assess gamma-aminobutyric acid receptor density and [3H]flunitrazepam or [3H]Ro 15-1788 was used to assess benzodiazepine receptor density. No significant differences were observed between the magnitude of binding of the three ligands to each of the microregions of brain from control rabbits and rabbits in Stage III or IV hepatic encephalopathy. These findings suggest that the behavioral expression of hepatic encephalopathy in the model studied is not dependent upon an increase in the number of gamma-aminobutyric acid or benzodiazepine receptors, but do not conflict with the hypothesis that gamma aminobutyric acid-ergic tone is increased in hepatic encephalopathy. PMID- 2545588 TI - Prolonged duck hepatitis B virus replication in duck hepatocytes cocultivated with rat epithelial cells: a useful system for antiviral testing. AB - Duck cultured hepatocytes from Pekin ducks naturally infected by duck hepatitis B virus can remain functional twice longer if a coculture system with rat liver epithelial cells is used instead of ordinary primary culture. The use of a selective medium in which ornithine and lactate replaced arginine and glucose, respectively, allowed viral replication initiated in vivo to be maintained in the coculture for 2 months. Several antiviral compounds including the pyrophosphate analog (phosphonoformic acid) or nucleoside analogs (9 beta-arabinofuranosyl AMP, 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-iodocytosine, 1,2'-deoxy-2' fluoro-beta-D-arabinofuranosyl-5-ethyluracil and 1,2'-deoxy-2'-fluoro-beta-D arabinofuranosyl thymine) were studied in both culture systems for their ability to inhibit duck hepatitis B virus replication. Hepatocytes were treated for 7 days with 1,2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl-5-ethyluracil (10 microM) and 1,2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl thymine (0.5 microM) or for 14 days with 9 beta-arabinofuranosyl AMP (90 microM), phosphonoformic acid (100 microM) and 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-iodocytosine (6 microM). The effects of the drugs on viral replication were monitored by testing for duck hepatitis B virus DNA in the culture supernatant and in the cells by molecular hybridization.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545589 TI - Effect of galactose on interaction of N-(2-hydroxypropyl)methacrylamide copolymers with hepatoma cells in culture: preliminary application to an anticancer agent, daunomycin. AB - A series of copolymers were prepared containing 1,2:3,4-di-O-isopropylidene-6-O methacryloyl-alpha-D-galactopyranose (0 to 99 mol %), methacryoyltyrosinamide and N-(2-hydroxypropyl)methacrylamide (99 to 0 mol %). The effect of galactose content on interaction with hepatoma cells in vitro was studied. Increased galactose content caused increased accumulation of N-(2 hydroxypropyl)methacrylamide copolymers by two human hepatoma cell lines (Hep G2 and SAH), but accumulation by rat and mouse hepatoma (HTC and NCTC) was not galactose dependent. Accumulation of N-(2-hydroxypropyl)methacrylamide copolymers by Hep G2 was shown to be an active process, being inhibited by low temperature and by the metabolic inhibitor 2,4-dinitrophenol. Addition of N acetylgalactosamine and polymer-galactose to the incubation medium resulted in a concentration-dependent inhibition of accumulation of galactose-containing polymers. Addition of fucose or galactose was without effect at the concentrations used. Polymers bearing galactosamine or fucosylamine residues and, in addition, daunomycin were evaluated for cytotoxicity against Hep G2 and SAH. N (2-Hydroxypropyl)methacrylamide copolymer-bound daunomycin produced a dose dependent inhibition of DNA synthesis (measured by incorporation of [3H]thymidine), and the galactose-containing polymer showed greatest inhibition. PMID- 2545590 TI - Characterization of woodchuck hepatitis virus DNA and RNA in the hepatocellular carcinomas of woodchucks. AB - Integration and transcription of woodchuck hepatitis virus DNA were studied by Southern and Northern blot analysis in 26 hepatocellular carcinomas and in adjacent nontumor tissue of woodchucks (Marmota monax). All liver tissue chronically infected with woodchuck hepatitis virus contained various amounts of episomal and replicative forms of woodchuck hepatitis virus DNA: episomal and replicative forms of woodchuck hepatitis virus DNA without integration were found in six tumors, episomal and integrated woodchuck hepatitis virus DNA was observed in 18 tumors and exclusively integrated woodchuck hepatitis virus DNA was found in two tumors. In most tumors and in all of the liver tissues chronically infected with woodchuck hepatitis virus, two major woodchuck hepatitis virus RNA species (3.7 and 2.1 kilobases) were detected. In tumors of two other animals (HW76 and HW89) with integrated woodchuck hepatitis virus DNA, only single major transcripts of 3.5 and 2.5 kilobases, respectively, were detected. Hybridization with subcloned woodchuck hepatitis virus DNA probes showed that both aberrant transcripts lacked the C gene and a part of the pre-S1 gene; characterization of corresponding integrated woodchuck hepatitis virus DNA sequences revealed that the C gene was deleted in one tumor, although not in the other. In agreement with the nucleic acid data, we found expression of core protein by Western blotting only in chronically infected liver tissue of these animals, but not in the corresponding tumors. Deletion of the C gene in mRNA may be due to deletion of this gene in the integrated sequences or due to transcriptional regulation. PMID- 2545591 TI - Carbohydrate metabolism in hepatocellular carcinoma: where does the glucose go? PMID- 2545592 TI - Pure and mixed mucinous carcinomas of the breast: a clinicopathologic analysis of 61 cases with long-term follow-up. AB - Sixty-one mucinous carcinomas (MCs) of the female breast were followed-up for at least 18 years or until death (mean follow-up time, 26 years; median follow-up time, 23 years). The 61 MCs were compared with 441 unselected cases of breast carcinomas of all histologic types (reference carcinomas or RCs), which were follow-up for at least 21 years. When the MCs were divided into pure (PMCs) and mixed (MMCs) mucinous carcinomas, the 20-year cumulative corrected survival rate for operable cases in the PMC group was 79% +/- 11% (SE) and 28% +/- 13% for the MMC group. The difference is statistically significant (P less than .001). The PMCs had a significantly better survival rate (P less than .001) when compared with the RCs (20-year corrected survival rate, 41% +/- 3%). The survival rates for the MMCs and RCs did not differ significantly from each other. By Cox's multivariate analysis, pure histologic type and a tumor size less than 5cm were independent favorable prognostic factors in the MC group, but nodal status was closely related to the histologic type. Judging from the relative survival curves, no significant excess mortality of cancer occurred toward the end of the follow-up period in the PMC group. PMID- 2545593 TI - Malignant giant cell tumor of synovium and locally destructive pigmented villonodular synovitis: ultrastructural and immunohistochemical study and review of the literature. AB - The first reported case of an intraarticular malignant giant cell tumor of synovium studied with electron microscopic and immunohistochemical examination is presented, together with a case of diffuse intraarticular pigmented villonodular synovitis with extensive bone destruction. The malignant case was dominated by uniform cells positive for histiocytic markers, the fine structure showing a gradual change from cells dominated by organelles serving a secretory function to cells with phagocytic activity. The reported cases of giant cell tumor of the tendon sheath indicate that the pertinent histologic changes regarding malignancy are an increase in cell polymorphism and in the number of mitoses, and a decrease in the number of multinucleated giant cells. PMID- 2545594 TI - Identification of Epstein-Barr virus-infected cells in tonsils of acute infectious mononucleosis by in situ hybridization. AB - In situ hybridization with 35S-labeled Epstein-Barr virus (EBV) probes was applied to paraffin sections of tonsils from seven patients with clinical, serologic, and morphologic evidence of acute infectious mononucleosis. EBV genomes were demonstrated in activated lymphoid B blasts in the interfollicular and perifollicular zones in all these cases. However, in no case could EBV be identified in epithelial cells. These results are at variance with the current concept which attributes a central role to the tonsillar epithelium in primary EBV infection. PMID- 2545595 TI - HPV and cervical cancer. PMID- 2545596 TI - Regional and physical mapping studies characterizing the Greig polysyndactyly 3;7 chromosome translocation, t(3;7)(p21.1;p13). AB - The Greig polysyndactyly-craniofacial anomalies syndrome is an autosomal dominant disorder involving a gene(s) located in band 7p13. We have isolated and characterized a reciprocal 3;7 chromosome translocation that resulted in the syndrome. We have identified two closely linked (0 cM) conserved DNA sequences (P137/p944B) that flank the translocation breakpoint. A pulsed-filed analysis combined with available genetic linkage information demonstrates that the disorder is linked (2 cM) to the T-gamma receptor locus, lending considerable support to the hypothesis that the mouse mutant extra-toes is the counterpart of the Greig syndrome. We have found no evidence that physically links the EGF receptor to the P137/p944B region, again compatible with mouse linkage relationships. The isolation of the der(3) chromosome from the 3;7 translocation has allowed us to regionally localize probes within the 3p21.1 band. For three probes commonly used in heterozygosity experiments with human cancers involving chromosome 3, we have determined that the order from centromere to telomere is D3S3, D3S2, and DNF15S2. Our pulsed-field studies also demonstrate the utility of band density differences combined with partial digests in evaluating linkage relationships. The P137/p944B probes should be useful in examining other hereditary disorders with phenotypic similarities to the Greig syndrome. PMID- 2545597 TI - Effect of homeopathic dilutions on subcellular enzymatic activity. AB - The activity of various inhibitors on several subcellular enzymes was studied. First we determined the inhibitory concentration required to reduce maximum enzymatic activity by 50%, then the effect of various hahnemannian dilutions of the same inhibitory agent was tested. Seven inhibitory agents were tested in this way on seven different enzymatic systems. No effects of these hahnemannian dilutions were shown. PMID- 2545598 TI - A live cell enzyme-linked immunosorbent assay for detecting human hepatoma membrane antigens. AB - Live cell enzyme-linked immunosorbent (ELISA) and fixed cell indirect immunofluorescence (IF) assays were compared to screen mouse hybridomas producing immunoreactive monoclonal antibodies against cell membrane antigens expressed on Ha22T, a human hepatoma cell line. While performing live cell ELISA, two parameters were tested to improve the viability of the target cells. The first parameter was the inclusion of growth medium in the assay buffers, and the second was performing the assay incubations at 37 degrees C in an incubator containing 5% CO2 in the air. Fixed cell IF detected and classified 46% of the hybridomas secreting monoclonal antibodies reactive with membrane, cytoplasm, cytoskeleton, and nuclear antigens of Ha22T cells. Fixed cell IF was able to reveal mixtures of two or more hybridomas growing in the same well secreting antibodies to different cell organelles. The live cell ELISA, on the other hand, identified 12 additional membrane reactive monoclonal antibodies from the hybridoma supernatants that were not reactive by IF. These results disclose that cell fixation procedures used for IF either completely or partially inactivated some of the cell membrane antigens. We, therefore, propose the use of a combination of immunoassays to select the maximum number of hybridomas secreting useful monoclonal antibodies from somatic cell fusions. PMID- 2545599 TI - Functional neuromuscular stimulation system using an implantable hydroxyapatite connector and a microprocessor-based portable stimulator. AB - For developing functional neuromuscular stimulation (FNS) orthoses, percutaneous implants made of sintered hydroxyapatite (HAp) and a microprocessor-based portable stimulator were devised, and related basic studies were carried out. First, a button-shaped HAp was percutaneously implanted up to 23 months in the skin of dogs, and 30 months in human subjects. The HAp was closely contacted with the skin tissue. Then, button-shaped HAp electrical connectors were percutaneously implanted in the skin of rabbits. Sufficient contractions of the calf muscle were elicited by electrical stimuli delivered to intramuscular electrodes through the connector. These results indicated that the HAp electrical connector could be clinically utilized as an implantable skin interface for FNS. The microprocessor-based stimulator capable of processing control command signals and delivering 16-channels of electrical stimuli was developed. Usefulness of the stimulator was demonstrated by applying it to flexor and extensor muscles of the thumb in a C5 quadriplegic patient. PMID- 2545600 TI - Distinct patterns of conservation of exons and introns of T-cell receptor beta chain constant region genes in subspecies of Mus. PMID- 2545601 TI - Production of oxygen radicals by the reduction of oxygen arising from the surface activity of mineral fibres. AB - According to certain hypotheses, the production of oxygen radicals within the biological medium (the phenomenon of oxidative stress) may play an important role in fibrosis and in certain steps of carcinogenesis. The mineral fibres of various materials are capable of participating in this phenomenon, owing to the reducing nature of their surface activity, so that OH. radicals can be produced from oxygen in 3 steps. The surface activity of inorganic materials which are insoluble or only very slightly soluble is due to the presence of electron donor active sites, generally linked to Fe2+ ions found in the neighbourhood of the surface. In biological systems, these sites may emerge on the surface as a result of the partial dissolution of the particle, the action of a biological reducing agent, the phenomenon of deposition on the surfaces or cation exchange. We have explored the reducing properties of the surfaces of a certain number of mineral fibres, in aqueous buffer medium, by electron paramagnetic resonance (EPR) measurement of the adduct with the radical-trapping agent 5,5'-dimethyl-l pyrrolidine-N-oxide (DMPO), produced from the radicals initially formed (OH. or R.). We have found certain fibres to be highly effective in producing radicals from dissolved oxygen (Canadian chrysotile, nemalite, freshly ground amphiboles) while others have little effect. The reducing activity of certain fibres may be markedly increased by prior treatment in the presence of a ferrous salt (as in the case of erionite) or by the addition of glutathione to the reaction medium (as in the case of UICC crocidolite). It is suggested that the carcinogenic activity of certain inorganic materials at the pulmonary level is the result of their surface reducing properties. These reducing properties may either be present at the time of inhalation or acquired in the biological medium. This hypothesis is not in conflict with the observation of the role of the dimensional characteristics of fibres in mesothelioma. PMID- 2545602 TI - Experimental data on in vitro fibre solubility. AB - Different types of natural and synthetic fibres have been subjected to systematic solubility tests in vitro in a physiological solution at 37 degrees C. Both closed-system and open-system experiments were carried out. Atomic absorption spectrometry of the filtered fluids showed characteristic differences of solubility. Plastic fibres are practically insoluble. In contrast to glass fibres, the solubility of asbestos fibres is low. Sepiolite and wollastonite are of moderate solubility. The results were confirmed by scanning electron microscopy. Kinetic studies and extensive solubility tests led to a new exponential expression which describes the dissolution process in a closed system better than the square-root time laws often used. Moreover, this exponential model provides a new method of distinguishing between different materials by means of their initial rates of dissolution. PMID- 2545603 TI - Comparative study of the effect of chrysotile, quartz and rutile on the release of lymphocyte-activating factor (interleukin 1) by murine peritoneal macrophages in vitro. AB - The release of lymphocyte-activating factor (LAF) into the medium of cultured mouse resident peritoneal macrophages was estimated from the effect of this medium on the proliferation of mouse thymocytes in the presence of phytohaemagglutinin. Untriggered macrophages released little LAF into their culture medium. Upon addition of UICC chrysotile A (25-50 micrograms/10(6) macrophages), LAF appeared in the medium, and release continued for at least 20 h. DQ12 quartz was a more potent inducer of LAF production, while rutile was nearly inactive. Although chrysotile and quartz caused cell damage (as estimated by the release of lactate dehydrogenase), it was established that LAF release was not attributable to leakage of preformed intracellular LAF. The lymphoproliferative activity detected in macrophage media was stable at 56 degrees C and nondialysable, which corresponds to the properties of interleukin 1. These biochemical observations are consistent with the non-specific stimulation of the immune system found in asbestotic and silicotic patients. PMID- 2545604 TI - Translocation of subcutaneously injected chrysotile fibres: potential cocarcinogenic effect on lung cancer induced in rats by inhalation of radon and its daughters. AB - Exposure to radon 222 and its daughters has been shown to induce lung cancer in rats. The cocarcinogenic effect of intrapleurally injected mineral fibres in rats which have previously inhaled radon has also been established. The aim of this work was to establish whether a similar process could be induced at a distance from the lungs by subcutaneous injection of chrysotile fibres. Three groups of animals were used: (1) 109 rats which inhaled radon only (dose: 1600 working level months (WLM]; (2) 109 rats given a subcutaneous injection in the sacrococcygeal region of 20 mg of chrysotile fibres after inhalation of the same dose of radon; and (3) 105 rats injected with fibres only. No mesotheliomas occurred in any of the 3 groups. The incidence of lung cancer was 55% in group 2, 49% in group 1 and 1% in group 3. Statistical analysis using Pike's model showed that the carcinogenic insult was slightly higher in group 2 than in group 1. Electron microscopy analysis of fibre translocation from the injection site showed that less than 1% of injected fibres migrated to the regional lymph-nodes and only about 0.01% to the lungs. After injection, the mean length of the fibres recovered in lung parenchyma increased with time, suggesting that short fibres are cleared by pulmonary macrophages whereas long fibres are trapped in the alveolar walls. Although the high tumour incidence observed in group 1 might have masked the cocarcinogenic effect induced by the fibres, it is possible that this effect can occur only at short distances.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545605 TI - Transplantation and chromosomal analysis of cell lines derived from mesotheliomas induced in rats with erionite and UICC chrysotile asbestos. AB - The histological lesions, chromosomal characteristics and transplantability of 6 erionite-induced and 7 UICC chrysotile-induced rat mesotheliomas are compared. The tumours were of 4 types: tubulopapillary, fibrosarcomatous, mixed fibrosarcomatous and tubulopapillary, and mixed fibrosarcomatous and chondrosarcomatous. Cell lines derived from these tumours displayed heterogeneous chromosome anomalies, but none were unique either to chrysotile or erionite treatment. Six of 7 erionite-induced and 4 of 6 UICC chrysotile-induced tumours had various anomalies of chromosome No. 1. When 7 cell lines were transplanted into syngeneic rats, all produced tumours that were pathologically similar to the original tumour, regardless of the route of injection. Cytogenetically, the cell lines derived from tumours after intrapleural transplantation resembled the injected cell line. The cytogenetic analysis of the cell lines derived from the tumours after subcutaneous transplantation is in progress. The induction period for transplanted tumours was 28-80 days. PMID- 2545606 TI - Toxicity of an attapulgite sample studied in vivo and in vitro. AB - Conflicting data are found in the literature concerning the carcinogenic potency of attapulgite. We tested the carcinogenic potency of French attapulgite in rats, and compared it with 2 chrysotile samples: Rhodesian UICC (Ch A) and short Canadian fibres (Ch C). The mean length of the fibres was 0.77 micron (attapulgite), 3.21 microns (Ch A) and 1.25 microns (Ch C). The mean diameter was 0.06 micron in the 3 samples. The particles (20 mg) in saline were inoculated into the pleural cavity of Sprague-Dawley rats allowed to survive for their full lifespan. The incidence rates of mesothelioma were: 0% (saline controls), 0% (attapulgite), 19% (ChC) and 48% (Ch A). In vitro studies were carried out using cultures of rat pleural mesothelial cells (RPMC). Attapulgite and Ch C did not modify cell growth except at high doses of 10 micrograms/cm2. Unscheduled DNA synthesis (UDS) was detected using [3H]thymidine incorporation in confluent RPMC (GoG1 arrested) and a scintillation method. UDS was stimulated with either Ch A or Ch C at doses ranging from 2 to 10 micrograms/cm2. In contrast, attapulgite did not significantly enhance [3H]thymidine incorporation at doses ranging from 2 to 20 micrograms/cm2. The results show that the attapulgite tested here had no carcinogenic potency. The in vivo and in vitro reactivity of the fibres used in this experiment might perhaps be related to the fibre size; however, other parameters may also be important. PMID- 2545607 TI - Effect of choline and mineral fibres (chrysotile asbestos) on guinea-pigs. AB - Both short-term and long-term, low and high doses of parenterally administered choline produce pathological lesions in lungs and lymph-nodes of rats and guinea pigs. Male guinea-pigs given choline by intraperitoneal injection, 40 doses of 50 mg of choline chloride, 5 days per week for 8 weeks, developed lung lesions consisting of peripheral nodules of small cells, neoplastic bronchiolar epithelium, carcinomatous lesions and changes in the pleural surface by the end of the experiment (680 days). In a second group, a single intraperitoneal injection of 15 mg of chrysotile asbestos given after 210 days of choline administration resulted in the early onset of pulmonary lesions at 570 days but there was, in addition, evidence of enhancement of cancerous lesions both in lung and in lymph-nodes at 570 days and 680 days as compared with choline alone. It is clear from the present experiment that the parenteral administration of carcinogenic mineral fibres (chrysotile asbestos) and availability of excess choline act synergistically in producing cancerous lesions in lungs and other organs. PMID- 2545608 TI - Cytotoxicity and carcinogenicity of chrysotile fibres from asbestos-cement products. AB - Fibres from weathered asbestos-cement products have little or no haemolytic activity, as compared with UICC chrysotile; this is probably the result of magnesium leaching during the weathering process. Weathered samples of asbestos cement are cytotoxic, but the release of lactic dehydrogenase (LDH) by guinea-pig alveolar macrophages caused by low and intermediate dust concentrations of UICC chrysotile is greater than that of such samples. The influence of serum is different as between UICC chrysotile and asbestos cement. In the former, LDH release by macrophages is enhanced, whereas it is reduced in the latter. Cytotoxicity is length-dependent in respect of LDH release from macrophages and proliferating cells, as well as cell proliferation. In all test systems, the sample from the unweathered core of an asbestos-cement plate is less toxic; only in the haemolysis system using an unbuffered solution can erythrocyte destruction be observed. This may be because this sample contains fewer single fibres than the others and because the specific surface of those fibres is smaller. The carcinogenicity of the weathered asbestos-cement chrysotile fibres is comparable to that of standard chrysotile fibres following intraperitoneal (i.p.) application. PMID- 2545609 TI - Qualitative and quantitative evaluation of chrysotile and crocidolite fibres with infrared spectrophotometry: application to asbestos-cement products. AB - Infrared (IR) spectrophotometry allows simple and rapid qualitative and quantitative evaluations of different types of asbestos, as well as of other inorganic particles. In particular, chrysotile and crocidolite have characteristic IR spectra, and optical density measurements in the 2710 nm band for chrysotile and the 12820 nm band for crocidolite permit the quantitative evaluation of each fibre either alone or in mixtures. IR spectra also provide information on changes in fibre structure and in chemical composition as the result, for example, of thermal treatment or acid leaching. The analytical method that we have developed can detect amounts as small as 0.1 mg of fibre in a 300-mg disk of potassium bromide using a low-cost IR spectrophotometer. The use of a Fourier transform IR spectrophotometer dramatically improves the sensitivity and selectivity. Computer-assisted analysis of spectra offers the possibility of reducing matrix interference and of comparing different spectra. The application of the IR technique to asbestos-cement products and insulating materials is described. PMID- 2545610 TI - Non-occupational malignant mesotheliomas. AB - The mineral content of the lungs from 84 cases of malignant pleural mesothelioma was estimated by electron microscopy and energy-dispersive X-ray analysis. These cases were chosen because the history of asbestos exposure was absent, indirect or ill-defined. The occupational exposures were classified according to the method of Zielhuis, and the results indicated that this classification is unnecessarily complicated. The chrysotile counts in the lungs from these mesothelioma cases were similar to those in controls and in a previous series of mesotheliomas in which the majority had had direct exposure to asbestos. Amphibole counts were intermediate between those in controls and in the previous series of mesotheliomas with direct asbestos exposure. These findings confirm those of previous studies indicating that amphiboles are more important than chrysotile in the causation of malignant mesothelioma. The results confirm that some mesotheliomas develop in the absence of asbestos exposure. PMID- 2545611 TI - Fibre content of lung in amphibole- and chrysotile-induced mesothelioma: implications for environmental exposure. AB - Using 9 pairs of exposure-period-matched shipyard and insulation workers (amphibole exposure) and chrysotile-industry workers (chrysotile exposure) with mesothelioma, and an additional 9 pairs of workers with asbestosis, we found that the chrysotile workers with mesothelioma had 400 times the median lung fibre burden of the shipyard and insulation workers with mesothelioma. Mesothelioma in the chrysotile workers was associated with a 3 times greater median fibre burden than asbestosis, whereas in the shipyard and insulation workers mesothelioma was associated with only 1/35 the median amphibole burden seen in cases of asbestosis. In the chrysotile workers, the tremolite:chrysotile ratio and the mean fibre sizes were the same for both mesothelioma and asbestosis cases. These data suggest that total fibre load is crucial to the induction of mesothelioma by chrysotile, and that this phenomenon requires, on average, as high a fibre burden as induction of asbestosis by chrysotile. By contrast, for amphibole exposure, mesothelioma appears at a much lower fibre burden than asbestosis. The fibre types appear to differ by at least two orders of magnitude in their potential for inducing mesothelioma. Estimates of risk from environmental exposure must take these differences into account. PMID- 2545612 TI - Fibre type and burden in parenchymal tissues of workers occupationally exposed to asbestos in the United States. AB - Lung tissues obtained from 53 asbestos-exposed workers, and one person exposed in a domestic setting, were studied. Amosite is the most prevalent fibre, occurring in 74% of the specimens. Amosite is always found in the lungs of insulation workers whereas chrysotile is found in only 50% of this population. Crocidolite has been detected in 24% of the lungs examined, but this increases to 40% in workers with shipyard histories. Exposure to chrysotile is widespread; the fibre has been observed in 61% of the tissues studied. Chrysotile occurs as the lone fibre in about 22% of the tissues examined, but tremolite is present in one-third of these. Fibre consumption data cannot be used as indices of exposure in the workplace; amphibole exposure appears to be product- and job-category-related. The assessment of risk to asbestos disease in the general population of the United States, exposed to chrysotile, should be based on appropriate chrysotile exposed cohorts. PMID- 2545613 TI - Relation of environmental exposure to erionite fibres to risk of respiratory cancer. AB - During the period 1979-1983, IARC, in collaboration with the Department of Chest Diseases of Hacettepe University in Ankara and the MRC Pneumoconiosis Unit in Penarth, conducted an epidemiological and environmental survey in 4 villages in central Turkey affected by a high incidence of mesothelial tumours. Recent data point to erionite, a zeolite fibre, as the most plausible etiological agent. From animal experiments, erionite appears to be the most powerful carcinogenic fibre so far known. During the study period, 17 pleural mesotheliomas and 7 lung cancer cases have been reported among the villagers. These cancer cases are analysed in relation to exposure to fibres. We assume exposure to occur from birth onwards and therefore consider duration of exposure equal to age. On this basis, the incidence of mesothelial and lung tumours is analysed in relation to age and cumulative exposure to fibres computed using the airborne fibre levels measured during the survey. PMID- 2545614 TI - Bilateral pleural plaques in Corsica: a marker of non-occupational asbestos exposure. AB - In north-east Corsica, asbestos outcrops are a source of environmental pollution, as assessed by airborne concentrations of chrysotile and tremolite that are significantly higher in the north-east than the north-west. For this reason we compared the frequencies of patients with asbestos pleural plaques in these 2 regions by checking the X-rays of 1721 patients born in north Corsica who had not undergone any occupational exposure to asbestos. The percentage of plaques was respectively 3.7% in the north-east, and 1.1% in the north-west (p less than 0.05). The relative risk of 3.3 demonstrates a strong relationship between environmental asbestos and the pleural plaques found in north-east Corsica. PMID- 2545615 TI - Mesothelioma in Cyprus. AB - For many years, the main source of asbestos in Cyprus was thought to be the chrysotile mine in the central mountains. When a woman, who had no connection with the mine, developed mesothelioma, it was surprising to discover tremolite asbestos bodies within her lung. However, further studies have shown that tremolite occurs as a contaminant within the chrysotile ore body. In this study we have shown that both chrysotile and tremolite can be found in domestic and environmental samples throughout the mountain region; in particular, numerous fine fibres of both materials are present in stucco. Preliminary radiological studies have shown pleural disease in the village population and 5 out of 13 known cases of mesothelioma have arisen in persons unconnected with the mine. This suggests an environmental contribution to asbestos-related disease on the island. PMID- 2545616 TI - Asbestos fibre content of lungs with mesotheliomas in Osaka, Japan: a preliminary report. AB - That crocidolite and amosite are both associated with the development of mesothelioma is now well established, but earlier studies have failed to find an excess of chrysotile in lungs with mesotheliomas as compared with the amounts in lungs of unaffected controls. In an attempt to clarify the importance of fibre type in tissue, an examination of a series of mesotheliomas is being undertaken in Osaka, Japan. A total of 23 mesotheliomas and 5 rejected cases reviewed by the Osaka Mesothelioma Panel were examined for the types of asbestos and semiquantitative fibre content by means of a transmission electron microscope equipped with energy-dispersive X-ray analyser. Asbestos fibres were detected in 19 of the 23 mesotheliomas (21 pleura, 1 pericardium, 1 peritoneum; 19 males, 4 females). Amphibole fibres were found in 13 cases. Five pleural and one peritoneal mesothelioma were found to have only chrysotile fibres. One female pleural mesothelioma with neighbourhood exposure had short chrysotile fibres. Among the 5 rejected cases, only one case with occupational exposure had both chrysotile and amosite fibres. A group of 17 controls were also examined and asbestos fibres were found in 5. Our data, while not definitive, suggest that mesotheliomas can be induced in humans, not only by crocidolite and amosite, but also by chrysotile, though possibly to a lesser extent. PMID- 2545617 TI - Correlation between lung fibre content and disease in East London asbestos factory workers. AB - The lungs from 36 former workers at an East London asbestos factory dying of asbestos-related disease were compared with lung tissue from 56 matched control patients operated on in East London for carcinoma of the lung. The severity of asbestosis and the presence of pulmonary carcinoma or mesothelioma of the pleura or peritoneum were correlated with an asbestos exposure index and with the type and amount of mineral fibre of the lungs. Asbestosis was associated with far heavier fibre burdens than mesothelioma. Moderate or severe asbestosis was more common among those with carcinoma of the lung than in those with mesothelial tumours. Crocidolite and amosite asbestos were strongly associated with asbestosis, carcinoma of the lung and mesothelial tumours, whereas no such correlation was evident with chrysotile or mullite. It is suggested that greater emphasis should be placed on the biological differences between amphibole and serpentine asbestos fibre. PMID- 2545618 TI - Mesotheliomas--asbestos exposure and lung burden. AB - The assessment of asbestos fibres in the lungs at post mortem in groups of mesotheliomas, groups occupationally exposed to asbestos, and controls has shown that all these groups contain significant levels of asbestos as a lung burden. The amounts in each group are dependent on the degree of past exposure, being highest in those cases with a known or extrapolated occupational exposure, less in those cases with recorded neighbourhood or environmental exposure, and less again in those cases with no evidence of exposure to asbestos and in controls. Relative risk estimates and the use of models developed for occupational situations do not provide good estimates of the relevance of environmental fibres in producing mesotheliomas in the general population. This may be the result of differences between the groups in their time periods of exposure and long-term elimination of asbestos from the lungs. The number of mesotheliomas that might be due to low-level environmental exposure to asbestos cannot be determined from lung contents alone, but an assessment based on detailed occupational histories from the Australian Mesothelioma Surveillance Program show that the problem is not one of great importance when compared with other public health issues. PMID- 2545619 TI - Modification of fibrous Oregon erionite and its effects on in vitro activity. AB - A sample of erionite, a fibrous zeolite, was modified by milling to reduce the number and length of the fibres and by extraction with cyclohexane. The in vitro activities of this mineral were found to depend on the presence of long fibres. The erionite contained fewer of these fibres than the UICC asbestos samples but, unlike these materials, erionite can cause the transformation of C3H 10T1/2 cells. Erionite did not increase the activities of benzo[a]pyrene in this cell transformation assay. The cytotoxic activities of both asbestos and erionite have a similar dependence on the number of long fibres. Extraction with cyclohexane did not affect the activity of erionite. PMID- 2545620 TI - Mechanisms of fibre-induced superoxide release from alveolar macrophages and induction of superoxide dismutase in the lungs of rats inhaling crocidolite. AB - Asbestos resembles the phorbol ester, 12-O-tetradecanoylphorbol 13-acetate (TPA), in its ability to elicit release of superoxide (O2-.) from rodent alveolar macrophages (AM) in vitro. In addition, superoxide dismutase (SOD), the antioxidant enzyme scavenging O2-, is increased in cultures of tracheobronchial epithelial cells and lung fibroblasts after exposure to either crocidolite or chrysotile asbestos. Our objectives here were to determine: (1) the chemical and physical properties of asbestos important in the generation of O2- from rat AM; and (2) the effects of O2- in comparison with asbestos on biosyntheses of collagen and non-collagen protein in rat lung fibroblasts in vitro. We were also interested in whether increased production of SOD occurred in the lungs of rats after inhalation of crocidolite asbestos. To determine whether O2- was elicited in response to a variety of asbestiform fibres, AM lavaged from Fischer 344 rat lungs were exposed in vitro to equivalent non-toxic amounts of crocidolite asbestos, erionite, Code 100 fibreglass, sepiolite, and their non-fibrous analogues, riebeckite, mordenite and glass particles. In addition, sized preparations of long (greater than 10 microns) and short (less than 2 microns) asbestos were introduced at identical concentrations to determine whether length of fibres is critical in O2- release. The amount of O2- released from AM in response to dusts was then determined by measuring SOD-inhibitable reduction of cytochrome C. All asbestiform fibres caused a significant (p less than 0.05) increase in generation of O2- from epithelial cells, whereas non-fibrous particles were less active at comparable concentrations. Experiments with long (greater than 10 microns) versus short (less than 2 microns) chrysotile showed that long fibres caused a more striking, dosage-dependent release of O2-. To determine whether O2- plays a role in the causation of fibrotic lung disease, rat lung fibroblasts were exposed to a biochemical generation system (xanthine xanthine oxidase) for O2- before quantitation of cell-associated collagen and non collagen protein at 24, 48 and 72 h thereafter. At the latter time periods, significant increases in total collagen per ng DNA were observed. In comparison with controls, the generation system for O2- also caused an initial decrease in synthesis of non-collagen protein followed by increases in synthesis of non collagen protein at 48 and 72 h.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2545621 TI - Brief inhalation of chrysotile asbestos induces rapid proliferation of bronchiolar-alveolar epithelial and interstitial cells. AB - Inhalation of asbestos fibres causes a progressive interstitial pulmonary fibrosis. To understand the basic cellular mechanisms which lead to this disease, we have studied the earliest proliferative events at the bronchiolar-alveolar regions of rats and mice exposed to chrysotile asbestos for 5 h. Animals were injected with tritiated thymidine 4 h prior to sacrifice at varying times ranging from immediately after cessation of exposure to one month post exposure. Light microscopic autoradiography showed that air-exposed control animals never had more than 1% of cells labelled. Rats and mice studied immediately after exposure also had normal numbers of labelled cells. However, between 12 and 48 h post exposure, asbestos-exposed animals exhibited up to 4-fold increases in the percentages of labelled epithelial and interstitial cells. Normal labelling returned by 8 days after exposure and was maintained through the one-month period studied. We conclude that inhalation of chrysotile asbestos induces rapid and highly significant increases in proliferation of epithelial and interstitial cells of the bronchiolar-alveolar regions where asbestos fibres were initially deposited. PMID- 2545622 TI - Reduced adherence to traumatized rat heart valves by a low-fibronectin-binding mutant of Staphylococcus aureus. AB - Isogenic strains of Staphylococcus aureus, differing in fibronectin binding, were constructed for studies of the contribution of fibronectin binding to the in vivo pathogenesis of staphylococcal disease. Mutagenesis of S. aureus 879R4S was accomplished by mating with Enterococcus faecalis FA378 that carried the transposon Tn918. Four low-fibronectin-binding mutants were identified that bound 24- to 35-fold less fibronectin than the parent strain did. A spectinomycin resistant strain, R4SSp, was transduced by a bacteriophage (80 alpha) lysate propagated on a low-binding mutant of 879R4S to produce R4SSp/1536, which bound less fibronectin, contained a single copy of the transposon, and grew on spectinomycin-containing medium. Using a rat model of endocarditis, we determined the distribution of S. aureus R4SSp and its transductant in normal and cardiac catheterized rats. Cultures of heart tissue showed that catheterized rats challenged with the fibronectin-binding parent strain had over 250-fold more organisms in the left heart than did rats challenged with the low-binding transductant. The ability to bind fibronectin had no effect on the number of S. aureus cells cultured from other tissues. These data suggest that the ability of S. aureus to bind fibronectin is an important factor in establishing adherence to damaged heart valves in vivo. PMID- 2545623 TI - Cloning and characterization of the structural gene for the class 2 protein of Neisseria meningitidis. AB - The class 2 protein of Neisseria meningitidis is the major outer membrane protein and a porin. A lambda gt11 bank of meningococcal chromosomal DNA was screened with monoclonal antibodies against gonococcal protein IB that cross-react with meningococcal class 2 protein. Three independent immunoreactive clones were isolated. DNA sequence analysis indicated that these clones included regions encoding the N-terminal portion of the class 2 protein. An oligonucleotide of 21 bases that was complementary to this sequence was synthesized and used as a probe for a second screening of the lambda gt11 bank. One of the positive clones isolated contained the complete gene, including the ribosome binding site, but lacked the promoter region. On the basis of the DNA sequence, a protein of 360 amino acids was predicted. Comparison of the predicted protein sequence with that of gonococcal protein I showed little homology in six regions constituting 29% of the total amino acids, while the remainder of the coding frame showed 81% homology of amino acid residues. The DNA homology in the immediate 5' and 3' noncoding sequences was very striking. Following the putative transcription terminator, the 3' DNA sequence contained a complex pattern of direct and inverted repeats having some similarity to the 3' sequence of the gonococcal protein IB gene and very close homology to a sequence located in the pilS1 region (R. Haas and T. F. Meyer, Cell 44:107, 1986), an area of the gonococcal genome containing silent pilin genes. PMID- 2545624 TI - Comparison of the abilities of different protein sources of iron to enhance Neisseria meningitidis infection in mice. AB - This study was done primarily to determine whether the previously observed specificity of the meningococcal transferrin and lactoferrin receptors for human proteins was maintained in vivo during meningococcal infection in mice. Preliminary experiments evaluating the choice of host strain, the age and sex of mice, and the growth conditions of the meningococci indicated that 45-day-old female Swiss Webster mice challenged with meningococci grown on low-pH, low-iron Mueller-Hinton agar plates were appropriate for this study. The comparison of transferrins and lactoferrins from different species demonstrated that only the human forms of these proteins were utilized by meningococci; there was significantly greater mortality among mice treated with iron-saturated human transferrin or lactoferrin (93 and 100%, respectively) than among those not treated or treated with iron-saturated bovine transferrin or bovine lactoferrin (0%). Provision of exogenous hemoglobin also resulted in increased mortality, although not as great as that observed with amounts of transferrin with equivalent iron content, which parallels the more effective utilization of transferrin and lactoferrin in in vitro growth experiments. In addition, unlike with transferrin and lactoferrin, there was no difference in utilization of human and bovine hemoglobin in vitro or in vivo. PMID- 2545625 TI - Purification of the Clostridium spiroforme binary toxin and activity of the toxin on HEp-2 cells. AB - The two components Sa (Mr, 44,000) and Sb (Mr, 92,000) of Clostridium spiroforme toxin were identified and characterized. Serological data permitted the identification of two groups of actin ADP-ribosylating clostridial toxins. The first consists of only C. botulinum C2. The second group includes spiroforme toxin, iota toxin of C. perfringens E, and an enzyme called CDT found in one strain of C. difficile, antibodies against which cross-react with all of the members of both groups. C. spiroforme toxin acted on cells by disrupting microfilaments by ADP-ribosylation of G actin. Toxicity was not blocked by 10 or 20 mM ammonium chloride and was only moderately inhibited by 30 mM NH4Cl. Inhibition of coated-pit formation in HEp-2 cells by potassium depletion strongly protected against the effect of C. spiroforme toxin. Toxicity was not blocked by incubation of HEp-2 cells and spiroforme toxin at 15 degrees C. These results suggest that this new binary toxin enters cells via the coated-pit-coated-vesicle pathway and might reach the cytoplasm at the same time as or before transfer to early endosomes. PMID- 2545626 TI - Structure and mapping of antigenic domains of protein antigen b, a 38,000 molecular-weight protein of Mycobacterium tuberculosis. AB - Only a limited number of proteins from Mycobacterium tuberculosis have so far been shown to possess species-specific epitopes as defined by monoclonal antibodies. One such protein is protein antigen b (Pab) of molecular weight 38,000, which binds the monoclonal antibodies HYT 28, HAT 2, HBT 12, HGT 3, TB 71, and TB 72. The gene encoding this protein was isolated from a lambda gt11 M. tuberculosis DNA library. The nucleotide sequence of the recombinant mycobacterial insert was determined, and an open reading frame of 374 amino acids was identified. The amino acid sequence exhibited 30% homology to a phosphate binding protein, PstS, from Escherichia coli. The pab gene was subcloned into pBR322 in conjunction with the lacZ gene, and deletions were obtained from the 3' end. The anti-Pab monoclonal antibodies were used to probe crude protein lysates of E. coli transformed with the deletion plasmids. The monoclonal antibodies showed two reactivity patterns; one group of antibodies were dependent on the presence of the ultimate 91 amino acids of the protein, whereas another group of antibodies recognized an antigenic domain located on the middle portion of the molecule. None of the antibodies bound to the N-terminal 117-amino-acid peptide. PMID- 2545628 TI - Identification of additional virulence determinants on the pYV plasmid of Yersinia enterocolitica W227. AB - This paper describes the mutagenesis of the pYV plasmid from Yersinia enterocolitica W22703 (serotype O:9) with Tn2507, a new element generating operon fusions. Analysis of the mutants allowed the identification of an additional Yop protein called Yop20 and the mapping of yop20, yop44, yop48, and lcrV, the gene encoding the V antigen. The last gene appeared to be part of an operon that also may contain yop37 and yop44. At 37 degrees C, mutants affected in this operon grew poorly, irrespective of the presence of Ca2+, or they even died in the presence of Ca2+. This operon is thus involved in the regulation by Ca2+, and we called it car, for Ca2+ regulation. It is presumably the Y. enterocolitica counterpart of the lcrGVH operon of Yersinia pestis. Transcription of yop20 and of the car operon was strongly regulated by temperature and only slightly by calcium. Hence, these genes behaved like the other genes of the yop regulon. Mutants affected in yop20 or in yop48 were markedly less virulent for the desferrioxamine-treated mouse than was the parental strain. Yop20 and Yop48 thus probably are Yersinia virulence factors. PMID- 2545627 TI - Phorbol myristate acetate modulates calcium ion-dependent superoxide anion generation induced by a monoclonal antibody raised against polymorphonuclear leukocytes. AB - We used a monoclonal antibody, YI 51, raised against human polymorphonuclear leukocytes (PMN) to induce superoxide anion (O2-) generation in cells. Although YI 51 alone played only a small part in inducing O2- generation in PMN, the amount of O2- generation induced in 5 X 10(5) PMN was 3.7 to 5.5 nmol/min when F(ab')2 fragments of rabbit anti-mouse immunoglobulin antibody were added as a cross-linking agent. This O2- -inducing activity was high compared with that of wheat germ agglutinin (WGA), insoluble immunoglobulin G immune complexes (IC), or phorbol myristate acetate (PMA). The binding of YI 51 and soluble immunoglobulin G IC to PMN was not reciprocally inhibitory, indicating that YI 51 does not interfere with ligand binding to the Fc receptor-binding site. In the absence of calcium ion (Ca2+), O2- generation induced by YI 51 decreased to 10 to 20% of that in the presence of Ca2+. In contrast, O2- generation in response to WGA, IC, or PMA under Ca2+-free conditions was not affected. When PMN were pretreated with low concentrations of PMA (10(-10) to 10(-9) M), the amount of O2- generation by the cells in response to YI 51 in Ca2+-free buffer was enhanced in a concentration-dependent manner. It also equaled the O2- generated by the cells in buffer containing Ca2+. In cells pretreated with PMA, the amount of O2- induced by WGA was enhanced two- to threefold over that in untreated cells. In contrast, there was no augmentation over untreated cells with stimulation by IC. These results suggest that YI 51, IC, and WGA induce O2- generation in human PMN in different manners. PMID- 2545630 TI - First observation of hopeite and parascholzite in fibrous capsules surrounding silicone breast implants. AB - We observed mineralization in 32.6% of the capsular tissue of 97 mammary prostheses recovered after surgical excision. Three distinct mineralogical phases were identified. The first was composed of common hydrated carbonate hydroxylapatite, a well-known physiological and pathological mineral. Hopeite and parascholzite constituted the second and third phases. They are zinc and calcium hydrate phosphates that have not been observed previously in human tissues. Statistical analysis of the data showed relationships between the presence of mineralizations and the manufacturers on the one hand, and the presence of Dacron fixation patches at the surface of the prostheses on the other. PMID- 2545629 TI - Loss of catalase activity in Tn1545-induced mutants does not reduce growth of Listeria monocytogenes in vivo. AB - Two catalase-negative mutants of Listeria monocytogenes were obtained by chromosomal insertions of the conjugative transposon Tn1545. The loss of catalase activity did not reduce the level of virulence of these mutants in mice. Indeed, both mutants were capable of growing in host tissues at the same rate as the parental catalase-positive strain. These results favor the view that catalase does not play a critical role in the resistance of L. monocytogenes to macrophage killing. PMID- 2545631 TI - Diet and colorectal cancer with special reference to fiber intake. AB - In this nested case-control study, 8,006 American Japanese men were examined and interviewed with a dietary questionnaire from 1965 to 1968. After a follow-up period of over 16 years, 102 colon and 60 rectal cancer incident cases were identified. Dietary data from these patients and from 361 cancer-free controls were analyzed for intake of dietary fiber (DF), vitamins, minerals, macronutrients, and selected food groups. We found a significant (p = 0.042) negative association of DF and colon cancer risk among low fat intake men (less than 61 g/d). In this subgroup, the men consuming less than 7.5 g/d of DF had an adjusted relative risk (RR) for colon cancer of 2.28 (95% CI 0.93-5.60), compared to those consuming greater than or equal to 14.8 g/d of DF. We also observed (among the complete group of subjects) a significant (p = 0.011) negative association between vitamin C intake and the risk of colon cancer. Men in the lowest quintile of vitamin C intake (less than 37 mg/d) had an adjusted colon cancer RR of 1.87 (95% CI 1.03-3.37), compared to men in the highest quintile (greater than or equal to 160 mg/d). We view these dietary associations with colon cancer risk with caution. There were no other significant associations of dietary variables with colon cancer risk. Also, there were no significant associations between intake levels of DF, micronutrients, or food groups and rectal cancer risk. PMID- 2545632 TI - Partial transcription map of Marek's disease herpesvirus in lytically infected cells and lymphoblastoid cell lines. AB - Marek's disease herpesvirus (MDV) can cause either a productive-restrictive or lytic infection, a latent infection or can transform thymus-derived lymphocytes. RNA was extracted from infected chicken embryo fibroblasts (CEF) or from lymphoblastoid tumour cell lines. Some of the infected CEF were treated with 200 micrograms/ml cycloheximide to identify immediate early (IE) transcripts, and others with 1 microM 1-(2-fluoro-2-deoxy-B-D-arabinofuranosyl)-5-methyluracil (FMAU), an inhibitor of herpesvirus DNA synthesis to identify early transcripts. An extensive Northern blot analysis was carried out using DNA probes spanning almost the complete MDV genome. In the lytically infected CEF at least 66 discrete transcripts were detected, ranging in size from 9.1 kb to 0.6 kb. Eleven IE transcripts were identified, of which 8 were mapped in the genome segment consisting of the IRL, IRS, US and TRS. Six transcripts were identified as early genes. In the MD lymphoblastoid cell lines MDCC-HPI, a non-producer cell line, and MDCC-CU41, a non-expression cell line, 4 and 7 transcripts were detected, respectively. These RNAs were transcribed from IE genes located mainly in the repeat sequences flanking UL and US and in US. Treatment of the lymphoblastoid cell lines with 20 micrograms/ml 5-iodo-2-deoxyuridine resulted in the additional transcription of 1 RNA species in HPI and 9 in CU41. Most of the transcripts present in lytically infected cells were also detected in MDCC-CU36, a cell line with a high percentage of antigen-positive cells (expression cell line). PMID- 2545633 TI - Expression of TGF-alpha/EGF and TGF-beta receptors in human colon carcinoma cell lines. AB - Previous studies have established that colon carcinoma cells secrete several polypeptide growth factors, including TGF-alpha/EGF and TGF-beta, suggesting that these and related molecules function in an autocrine/paracrine fashion to modulate tumor-cell growth. To investigate this possibility, we have studied the expression of transforming growth factor receptors in a panel of human colon carcinoma cell lines and in several untransformed epithelial cell populations. The results have revealed that neoplastic colon cells express receptors for both TGF-alpha/EGF and TGF-beta. Immunoprecipitation identified the TGF-alpha/EGF receptor as a structurally intact 170-kDa protein. No evidence for over expression was found. TGF-alpha (and EGF) enhanced receptor autophosphorylation, indicating that these receptors were biochemically functional. TGF-beta blocked DNA synthesis in non-neoplastic epithelial cells but not in tumorigenic colon populations. There was no correlation with TGF-beta receptor number or dissociation constant. However, chemical cross-linking studies revealed a TGF beta receptor subtype of 75 kDa in 3 of the 4 colon carcinoma cells which was undetectable in normal IEC epithelial cultures, suggesting a possible association between 75-kDa receptor expression and refractoriness to growth inhibition of TGF beta. Together, these data support the concept that locally-produced growth regulators can function in an autocrine or paracrine manner to influence the proliferation of colon carcinoma cells. PMID- 2545634 TI - Cytomegalovirus enhances lysis of HIV-infected T lymphoblasts. AB - A T4+ lymphoblastoid cell line (CR-10) persistently infected with the human immunodeficiency virus (HIV) and designated CR-10/NIT was superinfected with cytomegalovirus (CMV) isolated from peripheral blood mononuclear cells of a patient with AIDS. A productive CMV cycle in the CR-10/NIT lymphoblasts was demonstrated by fluorescent antibody staining (IF) using a monoclonal antibody (MAb) to the 150-kDa major capsid protein, by infectivity assays and by electron microscopy (EM). Two-color IF analysis showed that a small percentage of the CR 10/NIT cells were producing both CMV and HIV at any one time. EM studies revealed that all doubly infected cells were lysed whereas most cells infected only with HIV appeared intact. Cell lysis appeared 24 hr after superinfection of the CR 10/NIT cells with CMV and progressed to complete destruction of the cell culture between days 9 and 10. Our results suggest that CMV may convert a mildly cytopathic HIV infection of T lymphoblasts into a highly lytic process. PMID- 2545635 TI - Oncogene expression in small-cell lung cancer cell lines and a testicular germ cell tumor: activation of the N-myc gene and decreased RB mRNA. AB - N-myc and L-myc proto-oncogenes are expressed in many developing embryonic tissues. In contrast to expression of the closely related c-myc gene, N-myc and L myc expression is very restricted in adult tissues. We show that small amounts of the L-myc RNA can be detected in normal adult testicular tissue. A high level of N-myc expression from a single-copy N-myc gene was found in a malignant tumor of the testis, histopathologically defined as a seminoma. This tumor also showed a decrease of mRNA from the retinoblastoma gene (RB), which is ubiquitously expressed in all normal tissues. Strikingly similar results on elevated expression of the L-myc and N-myc genes and a lack of RB mRNA have been reported in small-cell lung cancer cell lines, and are confirmed in our study. PMID- 2545636 TI - Amplification and expression of different myc-family genes in a tumor specimen and 3 cell lines derived from one small-cell lung cancer patient during longitudinal follow-up. AB - In a small-cell lung carcinoma (SCLC) tumor specimen as well as in 3 cell lines derived from SCLC biopsies obtained from the same patient at successive times during the clinical course, either the N-myc gene or the c-myc gene appeared to be amplified and expressed. The initial tumor specimen, a lymph-node metastasis, was amplified for N-myc, as was the cell line GLC-14 derived from this metastasis. The cell lines GLC-16 and GLC-19, derived from the recurrent primary tumor biopsies after a complete remission, were amplified for c-myc. This finding implies independent amplification events and supports the idea that the amplification of myc genes is probably a secondary event correlated with tumor progression. Although all 3 cell lines could be classified as classic SCLC cell lines according to their histological characteristics, GLC-16 and GLC-19 clearly possess, in their c-myc amplification and derivation from therapy-resistant tumor cells, features of variant SCLC lines. This may question the significance of the classic/variant classification. PMID- 2545637 TI - The transforming prototype of Epstein-Barr virus (B95-8) is also a lytic virus. AB - The B95-8 isolate of the Epstein-Barr virus (EBV) has been described as a non lytic transforming virus. We have performed experiments in order to determine if the B95-8 EBV is capable of super-infecting and replicating in EBV-genome positive non-producer lymphoblastoid cells. Using concentrates of B95-8 EBV, prepared from 6 different B95-8 cell lines treated with 12-O-tetradecanoylphorbol 13-acetate (TPA), we demonstrated that virus concentrates could transform human or cotton-top tamarin B-lymphocytes and also lytically replicate in Raji cells, inducing EBV antigens and infectious virus. While the virus obtained from B95-8 super-infected Raji cells was able to transform cord-blood lymphocytes (CBLs) and super-infect Raji cells, transformation was abortive, with cell cultures only growing for up to 6 weeks. Transformation titers of the B95-8 virus concentrates ranged from 10(5) to greater than 10(8) transforming units/ml; early antigen (EA) induction ranged from 1% to 50% after superinfection of Raji cells, depending on the virus stock used, as determined by immunofluorescence. Southern blot analysis was carried out on the DNA prepared from B95-8 cells and virion DNA. The results were consistent with the published EcoRI restriction pattern for B95-8 EBV. The issue of whether the B95-8 cells produce virions with a dual biological phenotype or, rather, 2 biologically distinct viruses, is addressed. PMID- 2545638 TI - Methotrexate serum-level determinations during low-dose therapy of rheumatoid and psoriatic arthritis. AB - In 29 patients, 21 suffering from psoriatic arthritis and eight patients suffering from rheumatoid arthritis, methotrexate serum-levels were determined by means of radioimmunoassay. The aim of the investigation was to recognize an eventual dependence of the serum level of methotrexate on the total cumulative dose and to test the possibility of a concomitant therapy control. Beside the determinations of the serum levels of methotrexate, clinical examinations and laboratory tests were done at regular intervals. The values obtained showed no significant increase during the course of therapy compared to the values at the beginning of the treatment. Likewise no correlation to the total cumulative dose, the clinical picture or to the occurrence of side-effects could be found. Nor could any relationship between the changing of laboratory parameters and the methotrexate serum-levels be observed. No differences appeared in the methotrexate serum-levels during therapy of either rheumatoid or psoriatic arthritis patients. In conclusion it seems impossible to monitor a low-dose methotrexate therapy by continuous determinations of the serum levels of the drug. PMID- 2545639 TI - Antilipolytic effects of insulin and adenylate cyclase inhibitors on isolated human fat cells. AB - Antilipolysis induced by insulin by adenylate cyclase inhibitors was compared in isolated human fat cells when lipolysis was activated at well-defined steps in the cyclic AMP system. The latter was achieved with isoprenaline (beta adrenoreceptor agonist), cholera toxin and pertussis toxin (acting on the GTP sensitive coupling proteins), forskolin (stimulating the catalytic component of adenylate cyclase), enprofylline (selective phosphodiesterase inhibitor) and N6 monobutyryl-cyclic-AMP or 8-bromo cyclic-AMP (cyclic AMP analogues which are resistant or sensitive to phosphodiesterase, respectively). Clonidine (alpha 2 adrenoreceptor agonist), prostaglandin E2 and N6-(phenylisopropyl) adenosine (adenosine analogue) failed to inhibit lipolysis stimulated by cholera toxin or pertussis toxin, but were effective under all other conditions. Insulin failed to inhibit lipolysis stimulated by enprofylline or N6-monobutyryl cyclic AMP, but was effective under all other circumstances. In conclusion, insulin and adenylate cyclase inhibitors are antilipolytic in human fat cells through different mechanisms. Adenylate cyclase inhibitors act predominantly on the GTP-sensitive coupling proteins and, to a minor extent, at some yet unidentified distal step in the lipolytic machinery. As regards insulin, the major site of the antilipolytic action is phosphodiesterase. PMID- 2545640 TI - A dietary fibre supplement and weight maintenance after weight reduction: a randomized, double-blind, placebo-controlled long-term trial. AB - Ninety-seven mildly obese females (BMI = 27.4 kg/m2) were in a randomized, double blind, placebo-controlled trial treated for 52 weeks. The treatment consisted of a hypocaloric diet providing 5000 kJ/day (1200 kcal) and a dietary fibre supplement of 7 g/day for 11 weeks, (part I), followed by a diet providing 6720 kJ/day (1600 kcal) and a dietary fibre supplement of 6 g/day for 16 weeks (part II). Finally placebo was withdrawn and all still adhering subjects were given a dietary fibre supplement of 6 g/day and an ad libitum diet for the rest of the period (part III). Initial body weights were comparable, 76.9 +/- 0.8 kg in the fibre group versus 77.7 +/- 1.3 kg in the placebo group. During part I the weight reduction in the fibre group of 4.9 kg was significantly higher compared to that of 3.3 kg in the placebo group (P = 0.05). Accumulated weight reduction during part II was still significantly higher in the fibre group, 3.8 kg, compared to 2.8 kg in the placebo group (P less than 0.05). Total weight loss in the fibre group after 52 weeks was 6.7 kg. Probability of adherence to the treatment regimen was significantly higher in the fibre group from week 13 and onwards (P less than 0.01). Initial blood pressures were comparable. A significant reduction of systolic blood pressure occurred in both groups. A significant reduction of diastolic blood pressure occurred in the fibre group only, from 85.4 +/- 1.2 mmHg to 81.7 +/- 1.1 mmHg (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545641 TI - Effects of betaxolol/timolol on epinephrine stimulated cyclic-AMP levels in human trabecular meshwork cells. AB - The effects of timolol and betaxolol were compared for blocking beta agonist stimulation of cyclic-AMP in cultured human trabecular meshwork cells. Epinephrine (10(-5) M) produced a large and rapid increase in HTM cyclic-AMP; timolol (10(-6) M), at concentrations readily achieved in the aqueous humor after 0.5% eyedrops, completely blocked this effect. Recovery from timolol treatment appeared to be relatively slow, with only a 30-40% recovery observed by 9 hours. In comparison, betaxolol (10(-6) M) produced a smaller blockade of the epinephrine effects; a rapid recovery from the betaxolol effects was observed, with a greater than half-maximal response to epinephrine observed 15 minutes after removal of this beta blocker. These findings may help to explain the clinical observations of an outflow facility effect of epinephrine when used in combination protocols with betaxolol, but not with timolol. PMID- 2545642 TI - Transgenic animals. PMID- 2545643 TI - Herpes simplex retinitis in the mouse. Clinicopathologic correlations. AB - BALB/c mice inoculated with herpes simplex virus type 1 in one anterior chamber develop retinitis with subsequent retinal necrosis in the contralateral eye. The clinical features of this disease were characterized with indirect ophthalmoscopy and fundus photography. Three phases of the disease were delineated: (1) acute retinitis; (2) retinal necrosis; and (3) resolution; the clinical findings in each phase were correlated with the microscopic features. In addition, we compared the virus titers from individual animals with and without clinical evidence of retinitis to demonstrate that retinal inflammation correlated with the titer of virus in the uninoculated eye. This animal model is useful for the analysis of the clinical, virologic and immunologic manifestations of herpetic retinitis. PMID- 2545644 TI - Moderate Fuchs' endothelial dystrophy ATPase pump site density. AB - The Na+, K+-ATPase pump site density on corneal endothelial cells from Fuchs' endothelial dystrophy corneas has been shown to be drastically decreased in end stage disease (McCartney et al, Invest Ophthalmol Vis Sci 28:1955, 1987) and significantly increased in the early stages (Geroski et al, Ophthalmology 92:759, 1985) as compared to normal endothelium. In order to provide values for corneas between these two extremes, eye bank corneas from donors with no evidence of corneal edema but with guttata across the extent of the cornea were processed for autoradiography as well as immunohistochemistry. Pump site density was increased compared to end-stage disease but was less than values reported for either functional tissue or early stage disease. Similarly, immunohistochemistry results showed the amount of Na+, K+-ATPase antibody localization to be increased in respect to end-stage disease, but reduced as compared to functional tissue. These results suggest that pump site density on endothelial cells affected with Fuchs' endothelial dystrophy follows a gradual decline towards end-stage disease values as opposed to a sudden sharp deterioration after an initial increase. PMID- 2545645 TI - Inhibition of purified collagenase from alkali-burned rabbit corneas. AB - The inhibitory potency of four classes of compounds that inhibit corneal ulceration (thiols, tetracyclines, sodium citrate and sodium ascorbate) was assessed with collagenase purified from culture medium of alkali-burned rabbit corneas. The most potent inhibitor, a beta-mercaptomethyl tripeptide HSCH2(DL)CH[CH2CH(CH3)2]CO-Phe-Ala-NH2, exhibited 50% inhibition (IC50) at approximately 10 nM using the synthetic metalloproteinase substrate Dnp-Pro-Leu Gly-Leu-Trp-Ala-D-Arg-NH2. The inhibitor was somewhat less potent with type 1 collagen as substrate (IC50 between 1 and 3 microM), possibly because autooxidation of the essential - SH moiety of the inhibitor occurred during the longer time required for assay with the natural substrate. An N-carboxyalkyl tripeptide, CH3(CH2)2(DL)CH-(COOH)-Leu-Phe-Ala-NH2, was less potent (IC50 = 25 microM) than the thiol peptide. N-acetylcysteine, which is used to treat corneal ulceration, gave IC50 values of 2.7 mM and less than 10 mM with the synthetic and natural substrates, respectively. The IC50 values for the tetracyclines using the synthetic substrate were 15, 190 and 350 microM for doxycycline, minocycline and tetracycline, respectively. Inhibition by sodium citrate, but not the tetracyclines, could be reversed by excess Ca2+. Sodium ascorbate did not inhibit collagenase-mediated hydrolysis of either collagen or the synthetic substrate, thus indicating that the mechanism by which this agent inhibits corneal ulceration is not related to inhibition of collagen degradation by collagenase. PMID- 2545646 TI - Aldose reductase inhibitors and prevention of galactose cataracts in rats. AB - Our previous studies have shown that the aldose reductase inhibitor (ARI), sorbinil, prevents galactose-induced alterations and cataracts in rat lenses. We have now used sorbinil as well as another ARI, Eisai compound E-0722, to determine their potency in inhibiting aldose reductase- and galactose-induced alterations in lens morphology and Na+-K+-ATPase activity. Young Sprague Dawley rats were fed Purina Rat Chow plus 50% galactose, with or without 15 mg sorbinil, 0.15, 0.5 or 1.0 mg of E-0722/kg body weight per day. Controls were given Purina Rat Chow with or without ARIs. Lenses were studied for up to 60 days following the initiation of the diet using morphological, cytochemical and biochemical approaches to assess any alterations in the lens. While galactose-induced damage and cataracts were delayed by low doses (0.15 mg and 0.5 mg) of E-0722, they were completely prevented by the administration of 15 mg of sorbinil or 1 mg of E 0722/kg body weight per day. This study further showed that just 1 mg of E-0722 was more effective in preventing cataracts than 15 mg sorbinil. Thus it appeared that E-0722 was a more potent inhibitor of aldose reductase than sorbinil. PMID- 2545647 TI - Oxidative inhibition of Ca2+-ATPase in the rabbit lens. AB - Hydrogen peroxide inhibition of maximum Ca2+-ATPase and Na+,K+-ATPase activity was measured in a membrane-enriched preparation of rabbit lens cortical fibers and epithelium. At 5 X 10(-6) M hydrogen peroxide maximum Ca2+-ATPase activity was inhibited by 39%, while maximum Na+,K+-ATPase activity was stimulated. Ca2+ ATPase activity was almost completely inhibited at 5 X 10(-4) M hydrogen peroxide, in comparison to Na+,K+-ATPase activity, which was only inhibited by 28% at a concentration of hydrogen peroxide an order of magnitude larger. The addition of catalase to hydrogen peroxide-pretreated samples did not reverse the inhibition of Ca2+-ATPase by hydrogen peroxide. PMID- 2545648 TI - Receptor-mediated phosphoinositide hydrolysis in human ocular ciliary epithelial cells. AB - The hydrolysis of phosphoinositides (PI) in peripheral tissues can be stimulated by a number of putative neurotransmitters and this stimulation can be blocked by specific antagonists. Epithelial cells derived from the nonpigmented layer of the ocular ciliary epithelium were transfected by simian virus 40 and grown in culture to semiconfluency. The cells were incubated in 3 microCi/ml of (3H) myoinositol for 2 days. The accumulation of inositol phosphates in response to several agonists (carbachol, 1 mM; ATP, 100 microM; arginine vasopressin, 1 microM; and phenylephrine, 100 microM) was determined for times ranging from 5 sec to 15 min. In the presence of 10 mM LiCl, the maximum net production of the (3H)-inositol phosphates (expressed as a percent of conversion of (3H) phospholipids) was approximately 7.5% for inositol-1 phosphate, 0.5% for inositol 1,4 bisphosphate, and 1% for inositol-1,4,5 trisphosphate. Carbachol elicited PI hydrolysis with an EC50 value of 39 +/- 9 microM. The EC50 values obtained for arginine vasopressin and ATP-initiated PI breakdown were 32 +/- 10 nM and 11.9 +/ 1 microM, respectively. Phenylephrine alone failed to stimulate the production of (3H)-inositol phosphates in these cells. The production of all (3H)-inositol phosphates in response to carbachol (1 mM) was inhibited by atropine (Ki = 0.3 nM) and the selective muscarinic antagonists 4-DAMP (Ki = 4.2 nM), pirenzepine (Ki = 102 nM) and AFDX-116 (Ki = 1.49 microM). Thus the muscarinic receptors that are coupled to PI hydrolysis in these cells have the pharmacologic characteristics of the M3 subtype.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545649 TI - Heart-lung transplantation. AB - Since the successful clinical reintroduction of heart and lung transplantation in 1981, more than 350 of these procedures have been performed worldwide. Although survival following this operation is less than that reported for heart transplantation, the results are improving. It is clear that the increased technical difficulty of the procedure combined with the exquisite susceptibility of the transplanted lung to postoperative injury from infection, rejection or other causes account for these differences. In this report we provide an overview of the experience in heart-lung transplantation and discuss recent advances. The late complication of chronic obliterative bronchiolitis, which may progress inexorably, has cast a shadow over the potential long-term success of this therapeutic procedure. Current research efforts are directed toward the cause, diagnosis, and treatment of this complication. PMID- 2545650 TI - Free perforation due to colonic adenocarcinoma in Crohn's disease. AB - Free perforation of the bowel due to cancer in Crohn's disease is very rare. One case found among 75 cases of Crohn's disease is reported. The perforation occurred in a mucinous adenocarcinoma of the ascending colon in a 54-year-old woman with a 17-year history of Crohn's disease, who had undergone two previous operations of small bowel and colonic resection. A right hemicolectomy was performed; the patient had an uneventful recovery and does not show recurrence of cancer at 31-month follow-up. This is in contrast with a recent review, in which a very poor prognosis was claimed for the perforated colonic cancer in Crohn's disease. PMID- 2545651 TI - Carcinoma of the male breast and ocular metastases. AB - Breast carcinoma in males associated with ocular metastases is not often observed. A case is reported with review of the literature. Medical management is satisfactory in case of metastases and enucleation is suggested for severe pain. PMID- 2545652 TI - Di-octyl phthalate induced altered host resistance: viral and protozoal models in mice. PMID- 2545653 TI - Post-irradiation lactation. PMID- 2545654 TI - Nature of alpha subunit secretion in men: circadian rhythms, pulsatile release and secretory profiles. AB - Alpha subunit complements LH as a marker of the activity of the hypothalamic GnRH pulse generator. To characterize episodes of alpha subunit release and to determine if a circadian pattern of alpha subunit secretion is present in man, spontaneous alpha subunit pulsatility was analyzed in six healthy young men by blood sampling every 5 min for 24 h. The resulting alpha subunit concentration time series were analyzed by two statistically based independent peak detection methods, and subjected to Fourier transformation to assess underlying circadian rhythms. Cross-correlation analyses and multiple parameter deconvolution were used to estimate the concordance of spontaneous and exogenous GnRH-stimulated LH and alpha subunit secretion. These analyses revealed that two independent discrete peak detection algorithms yielded similar estimates of spontaneous alpha subunit pulse frequency, namely, 21 +/- 1.1 (Cluster) and 21 +/- 1.5 (Detect) alpha subunit peaks/24 h. Sampling intensity markedly influenced the estimate of endogenous alpha subunit pulse frequency, inasmuch as estimates from 5-min sampling were significantly greater than those of 10-min or 20-min sampling. Fourier transformation unmasked a significant circadian alpha subunit rhythm in all six men, with maximal concentrations at 0836 h and an average amplitude of 28% of the 24-hr mean hormone concentration. Cross-correlation analysis of spontaneous glycoprotein release revealed that serum LH and alpha subunit concentrations were highly cross-correlated when considered simultaneously, but not at various lags. Finally, deconvolution analysis of exogenous GnRH-stimulated glycoprotein release disclosed distinct half-times of alpha subunit and LH clearance with virtually simultaneous underlying secretory bursts. These data indicate that human alpha subunit is secreted in both a circadian and a discrete pulsatile fashion at a pulse frequency that is significantly underestimated at conventional sampling rates. The approximately hourly alpha subunit interpulse interval (68 +/- 4.6 min) is similar to that reported earlier for LH in peripheral blood and for testosterone in gonadal vein blood in healthy men. Moreover, cross-correlation analysis of endogenous GnRH-driven alpha subunit and deconvolution analysis of exogenous GnRH-stimulated alpha subunit and LH secretion suggest that these glycoproteins are secreted virtually simultaneously, but have significantly different endogenous clearance properties. The remarkably similar in vivo pulse frequencies for alpha subunit, LH, and testosterone in man suggest that the release of these three hormones is coordinately regulated. PMID- 2545655 TI - Effects of itraconazole on mitogen stimulated human mononuclear leucocytes. AB - Lymphocyte transformation responses of unstimulated and mitogen-stimulated cells from normal subjects were measured in vitro in the presence of itraconazole, at therapeutic and at much higher concentrations. No differences in response from those of control cell cultures were detected at any concentration. PMID- 2545656 TI - Susceptibility of penicillin-sensitive and -resistant strains of Streptococcus pneumoniae to new antimicrobial agents, including daptomycin, teicoplanin, cefpodoxime and quinolones. AB - The minimal inhibitory concentrations (MICs) of nine antibiotics were determined by agar dilution on 123 strains of Streptococcus pneumoniae (65 penicillin sensitive, 42 intermediate resistant and 16 resistant). The antimicrobial agents tested were penicillin G, clindamycin, trospectomycin, daptomycin, teicoplanin, cefpodoxime, ciprofloxacin, ofloxacin and vancomycin. Of these, daptomycin, teicoplanin and vancomycin demonstrated the greatest in-vitro activity against penicillin-resistant strains (MIC90s less than or equal to 0.25 mg/l). Ciprofloxacin, ofloxacin and trospectomycin had equivalent activities unaffected by penicillin-susceptibility (MIC90 of both quinolones 2.0 mg/l, and of trospectomycin 4.0 mg/l). Cefpodoxime was also active in vitro against all strains (MIC90 2.0 mg/l), but MICs increased with increasing penicillin-MICs. Most penicillin-susceptible strains were susceptible to clindamycin, but many penicillin intermediate resistant and resistant strains were resistant to this drug. Results of this study indicate that several newly introduced and experimental antibiotics have potential in the treatment of infections caused by resistant strains of Str. pneumoniae. PMID- 2545657 TI - Heterozygotes for cystic fibrosis: models for study of airway and autonomic reactivity. AB - Airway reactivity to cold air and methacholine, alpha-adrenergic and cholinergic reactivity measured as pupillary responses to phenylephrine and carbachol, respectively, and beta-adrenergic reactivity assessed by lymphocyte adenosine 3',5'-cyclic monophosphate (cAMP) response to isoproterenol were compared in 108 parents of patients with cystic fibrosis (CF) and 133 healthy adult controls. No differences were found between CF parents and controls in airway response to cold air or methacholine or in lymphocyte cAMP response to isoproterenol. Significant differences were found, however, in the response of the pupils to both phenylephrine and carbachol. Heterozygotes for CF have more reactive pupils; i.e., they require smaller doses of agonist for a 10% change in pupil size. In control subjects, the response of the pupils to phenylephrine and carbachol is highly correlated (r = 0.45, P less than 0.001), whereas in CF heterozygotes, the correlation is not significantly different from zero (r = -0.02). In controls, the pupil response to carbachol has a significant negative correlation with cold air response (r = 0.39, P less than 0.05), indicating that those whose pupils were most sensitive to carbachol had the greatest airway reactivity to cold air, but in CF heterozygotes the correlation is not significant (r = 0.10). A significant correlation exists between lymphocyte cAMP response and airway cold air response in CF heterozygotes (r = -0.32, P less than 0.05) (those whose beta adrenergic responsiveness is low have greater airway reactivity), but not in controls. The CF parents with the most reactive airways tend to have lower beta adrenergic responses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545658 TI - ACTH secretion and ventilation increase at similar arterial PO2 in conscious rats. AB - To compare the arterial PO2 (PaO2) at which adrenocorticotropic hormone (ACTH) secretion and ventilation are stimulated, conscious rats with chronic femoral arterial catheters were exposed for 50 min to 21, 18, 15, 12, or 9% O2. Decreases in arterial PCO2 (PaCO2) and increases in arterial pH and adrenocortical system activity occurred consistently throughout the exposure period in rats exposed to 9 or 12% O2. In contrast, changes in PaCO2 or pH were only transient or delayed, plasma ACTH did not change, and plasma corticosterone only increased after 20 min in rats exposed to 15 or 18% O2 relative to those breathing 21% O2. Omitting the large blood sample at 20 min for ACTH eliminated the increase in corticosterone in the 15% O2 group. Overall, ACTH increased, and PaCO2 decreased, below PaO2 of approximately 60 Torr. We conclude that ACTH secretion increases at a similar PaO2 as hyperventilation-induced decreases in PaCO2 and thus represents a primary physiological response to acute hypoxia; hemodynamic stimuli may also interact with hypoxia to augment adrenocortical system activity. PMID- 2545659 TI - Beta-adrenergic receptors are not responsible for exercise stimulation of glucose transport. AB - This study was designed to examine whether the increased glucose transport after acute exercise in rat skeletal muscle is mediated via beta-adrenergic receptor stimulation. Sarcolemmal (SL) membranes were isolated from three groups: control (C), acute exercise (E), and exercise + propranolol (E+P). The acute exercise bout was performed on a treadmill and consisted of a 45-min run until near exhaustion. E+P received an intravenous propranolol injection (0.8 mg/kg) 10 min before the exercise session. Michaelis-Menten kinetics at 1.5 s indicated that the Vmax for glucose transport was increased after each perturbation compared with C but were not different from each other (E, 4,334 +/- 377; E+P, 4,824 +/- 357; and C, 1,366 +/- 124 pmol.mg protein-1.s-1). The apparent Km's were similar in all groups. Scatchard plots for the D-glucose inhibitable class of cytochalasin B binding sites indicated no differences in either the total number of binding sites in the SL vesicles (C, 5.5 +/- 0.3; E, 5.1 +/- 0.2, and E+P, 5.1 +/- 0.3 pmol/mg protein) or in their dissociation constant (Kd) (C, 46 +/- 3; E, 48 +/- 3; and E+P, 49 +/- 2 nM). The increase in Vmax for transport was similar between E and E+P, indicating that exercise does not stimulate glucose transport via the beta-adrenergic receptor. PMID- 2545660 TI - Effect of rehydration on atrial natriuretic peptide release during exercise in the heat. AB - In an attempt to investigate their relationships with plasma volume (PV), heart rate (HR), and other hormonal systems, plasma atrial natriuretic peptide (ANP) levels were determined in response to exercise in the heat, associated with dehydration and rehydration with various fluids. Five normal subjects underwent four 3-h experiments, in a 36 degree C environment, in which 25-min exercise periods on a cycle ergometer at 90 W alternate with 5-min rest periods. Blood samples were collected hourly and ANP, arginine vasopressin (AVP), adrenocorticotropin (ACTH), and cortisol were analyzed in four experimental sessions: without fluid supplement (DH) and with progressive rehydration either with water (W), acid isotonic solution (AISO), or neutral isotonic solution (NISO). Exercise in the heat, accompanied by a decrease in PV and an increase in osmolality, elicited an increase of 28 +/- 1.6 pg/ml in plasma ANP, with concomitant increases in AVP (5.1 +/- 1.4 pg/ml), ACTH (49.6 +/- 12.3 pg/ml), and cortisol (8.4 +/- 2.0 micrograms/100 ml). Progressive rehydration maintained PV and blunted ANP, AVP, ACTH, and cortisol responses. These results demonstrate the importance of rehydration, during exercise in a warm environment, in preventing hormonal increases. They suggest that under our conditions, the PV changes and the inferred atrial pressure changes may not be the primary factors controlling ANP release, as under other physiological conditions. The exercise-related activation of pituitary and adrenals and the stimulation of HR counteract the influence of PV changes due to vascular fluid shifts. PMID- 2545661 TI - Plasma protein binding and endothelial enzyme interactions in the lung. AB - The influence of plasma albumin binding of the synthetic angiotensin-converting enzyme (ACE) substrate [3H]benzoyl-phenylalanyl-alanyl-proline (BPAP) on BPAP hydrolysis by pulmonary endothelial ACE was studied in isolated rabbit lungs perfused with a salt solution containing either 5% bovine serum albumin (BSA) or 5% dextran. The single-pass indicator-dilution method was used to measure the fraction (M) of [3H]BPAP hydrolyzed. Lung M was greater with albumin-free perfusate than when BSA was present. M decreased as the time (ti) that the BPAP was in contact with the BSA before reaching the lung was increased, suggesting that some BSA binding sites for BPAP were not in equilibrium during bolus transit through the lungs. The M vs. ti data were correlated using a model incorporating both rapid and slow binding kinetics of BPAP and BSA. For the slow BPAP-BSA interaction, the dissociation rate constant was approximately 0.015 s-1, and the fraction of the BPAP bound to these slowly equilibrating sites at equilibrium was approximately 22%. The results indicate that transient plasma protein binding kinetics can affect lung BPAP hydrolysis. PMID- 2545662 TI - Neutral endopeptidase and neurogenic inflammation in rats with respiratory infections. AB - Neuropeptides such as substance P are implicated in inflammation mediated by sensory nerves (neurogenic inflammation), but the roles in disease of these peptides and the peptidases that degrade them are not understood. It is well established that inflammation is a prominent feature of several airway diseases, including viral infections, asthma, bronchitis, and cystic fibrosis. These diseases are characterized by cough, airway edema, and abnormal secretory and bronchoconstrictor responses, all of which can be elicited by substance P. The effects of substance P and other peptides that may be involved in inflammation are decreased by endogenous neutral endopeptidase (NEP; also called enkephalinase, EC 3.4.24.11), which is a peptidase that degrades substance P and other peptides. In the present study, we report that rats with histories of infections caused by common respiratory tract pathogens (parainfluenza virus type 1, rat corona-virus, and Mycoplasma pulmonis) not only have greater susceptibility to neurogenic inflammatory responses than do pathogen-free rats but also have a lower activity of NEP in the trachea. This reduction in NEP activity may cause the increased susceptibility to neurogenic inflammation by allowing higher concentrations of substance P to reach tachykinin receptors in the trachea. Thus decreased NEP activity may exacerbate some of the pathological responses in animals with respiratory tract infections. PMID- 2545663 TI - Incorporation of arachidonic acid into lipids of the isolated perfused rat lung. AB - This study has attempted to identify the cells and phosphoglyceride molecular species associated with the rapid turnover of arachidonic acid (AA) in the isolated rat lung. In initial studies, AA complexed to trace amounts of albumin was added to the perfusate of rat lungs for 15 min and the incorporation of [3H]AA into various cells and phosphoglyceride molecular species was determined. Autoradiographic analysis revealed that the AA had labeled endothelial cells but also had already escaped from the intravascular space and labeled epithelial cells including alveolar type II cells. In addition, [3H]AA was found to be incorporated into various phosphoglycerides: phosphatidylcholine (PC) greater than phosphatidylethanolamine (PE) greater than phosphatidylinositol (PI). The majority of this [3H]AA was incorporated into 1-acyl-2-arachidonoyl-sn-glycero-3 PC, -PE, and -PI during the 15-min labeling period. In subsequent experiments, AA remodeling in the lung was examined by pulse labeling with [3H]AA for 15 min, washing unbound AA with albumin, and perfusing for an additional 120 min. In these lungs, some of the [3H]AA was remodeled into 1-alk-1-enyl-acyl-sn-glycero-3 PE. Gas chromatography-mass spectrometry analysis revealed that the largest pools of endogenous AA in the lung are found in PE associated with 1-alk-1-enyl-linked molecular species. On ionophore stimulation of lungs labeled for 15 min, labeled leukotriene (LT) B4, leukotriene C4, and 6-ketoprostaglandin F1 alpha (6-keto PGF1 alpha) were produced. LTC4 had a profoundly different radiospecific activity compared with LTB4 and 6-keto-PGF1 alpha, suggesting a different source of AA as contributing to the production of this eicosanoid.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545664 TI - Exercise training attenuates the myocardial dysfunction induced by endotoxin. AB - The purpose of this study was to determine whether exercise training protected against endotoxin-induced myocardial dysfunction. After a 12-wk treadmill training period, carotid catheters were implanted 24 h before saline or endotoxin administration into four groups of animals: trained saline-injected (TS), trained endotoxin-injected (TE), sedentary saline-injected (SS), and sedentary endotoxin injected (SE). Heart rate and mean arterial pressure were monitored 4 h after in vivo endotoxin or saline injection. Mean arterial pressure decreased an average of 32 +/- 3 mmHg 1 h after endotoxin administration but was normal (109 +/- 6 mmHg) 2 h later. Plasma catecholamines, in vitro myocardial performance, and isolated myocyte adenosine 3',5'-cyclic monophosphate (cAMP) production in response to isoproterenol were assessed 4 h after endotoxin injection. Plasma catecholamine levels were 5- to 15-fold higher in SE compared with the other groups. These data suggest that myocardial protection may be related to the lowered catecholamine levels elicited in TE compared with SE in response to endotoxin administration. The product of cardiac output and peak systolic pressure, an index of cardiac work, was 24-32% greater in TS compared with SS. Cardiac work was decreased 32% in TE compared with a 45% decrease in SE. cAMP was reduced in myocytes from SE in response to isoproterenol (-28%) and to forskolin (-44%) but not in myocytes from TE, compared with TS and SS. The difference in cAMP accumulation suggests that training maintains the integrity of the beta adrenergic receptor adenylate cyclase system, which can be depressed by in vivo endotoxin administration. PMID- 2545665 TI - The effects of omega-3 fatty acids on thrombosis and atherogenesis. PMID- 2545666 TI - Acute myeloid leukemia, FAB M-1 microgranular variant: a multiparameter study. AB - Acute leukemia was diagnosed in 62 adults and children over a recent 13-month period. Using light microscopy, cytochemical profiles, surface markers, and cytogenetics, 25 cases were classified as acute myeloid leukemia (AML) and 32 as acute lymphoblastic leukemia (ALL). The remaining 5 cases of de novo acute leukemia were unclassifiable. The routine cytochemical battery used on these 62 cases included: myeloperoxidase, sudan black B, nonspecific esterase, and periodic acid-Schiff (PAS). Flow markers utilized were: T3, T4, T5, T8, T10, T11, B1, B4, kappa, lambda, Ia, CALLA, Mo1, Mo2, My4, My7, My8, and My9. TdT was performed by immunoperoxidase and ELISA methods. The five unclassified cases were cytochemically negative and expressed no B- or T-cell-specific antigens, or TdT positivity. The morphologic differential diagnosis was between FAB L-2 and M-1. Karyotypic abnormalities involving chromosomes 3 and 7 were suggestive of myeloid origin in 2 of 4 patients studied. Flow cytometry demonstrated My7 on greater than 50% of blasts from two cases. Myeloperoxidase ultracytochemistry showed reaction product in small primary granules of blasts from all 5 cases. Positive cells contained only 1-2 granules/cell profile. The number of positive cells per case was in the range 10-20%. We conclude from this study that ultracytochemistry is very useful in providing definitive diagnosis and accurate subclassification of some AML FAB M-1 cases, particularly when light microscopic cytochemistry, cytogenetics, and flow cytometric markers are noncontributory. We propose to designate these acute "unclassified" leukemias as AML FAB M-1 "microgranular" type. PMID- 2545667 TI - Determination of water-insoluble cell walls in feeds: interlaboratory study. AB - A collaborative study was conducted to test a new rapid procedure for determination of water-insoluble cell wall (WICW) content in feeds. In the method, starch is solubilized near boiling temperature with Termamyl, a heat stable alpha-amylase, and proteins are solubilized at 40 degrees C with sodium dodecylsulfate and Pronase. Then, the organic matter of the residue is determined by incineration. Three hours were required to treat 12 different samples, including solubilization treatments, filtrations, and rinses. Eleven unknown products including 9 common feedstuffs of various origin and 2 mixed diets for poultry were analyzed by 7 analysts in France. Coefficients of variation ranged from 2.3 to 6.1%. The results were compared to those for water-insoluble dietary fiber (WIDF), total dietary fiber, and neutral detergent fiber. Agreement was best with the water-insoluble dietary fiber procedure. For most samples, the ratios of WIDF/WICW ranged from 0.981 to 0.842. The differences between WICW and WIDF values correspond to cell wall protein which is accounted for in WICW, but not in WIDF. PMID- 2545668 TI - Analysis of the H+/sugar symport in yeast under conditions of depolarized plasma membrane. AB - H+/sugar symport in the obligatory aerobic yeast Rhodotorula glutinis was analyzed under conditions where the plasma membrane was selectively depolarized by the lipophilic cation tetraphenylphosphonium (TPP+). Control experiments showed that this treatment did not impair the transmembrane delta pH, the cell energy charge, and the function of plasma membrane H+-ATPase. The kinetic data were fitted to elementary functions derived from a model constructed on the basis of some simplifying premises for ordered (either C + H+ + S or C + S + H+) and random reaction mechanisms. In addition, the comparison of the kinetic parameters in fully energized and depolarized cells provided information about the free carrier charge. It was concluded that the binding sequence of formation of the ternary carrier/H+/substrate complex follows a random mechanism and that the carrier bears a negative charge. PMID- 2545669 TI - Extensive Ca2+ release from energized mitochondria induced by disulfiram. AB - The effect of the alcohol-deterrent drug, disulfiram, on mitochondrial Ca2+ content was studied. Addition of this drug (20 microM) to mitochondria induces a complete loss of accumulated Ca2+. The calcium release is accompanied by a collapse of the transmembrane potential, mitochondrial swelling, and a diminution of the NAD(P)H/NAD(P) radio. These effects of disulfiram depend on Ca2+ accumulation; thus, ruthenium red reestablished the membrane delta psi and prevents the oxidation of pyridine nucleotides. The binding of disulfiram to the membrane sulfhydryls appeared to depend on the metabolic state of mitochondria, as well as on the mitochondrial configuration. In addition, it is shown that modification of 9 nmol -SH groups per mg protein suffices to induce the release of accumulated Ca2+. PMID- 2545670 TI - Cytochrome c oxidase of Euglena gracilis: purification, characterization, and identification of mitochondrially synthesized subunits. AB - Cytochrome c oxidase was purified from mitochondria of Euglena gracilis and separated into 15 different polypeptide subunits by polyacrylamide gel electrophoresis. All 15 subunits copurify through various purification procedures, and the subunit composition of the isolated enzyme is identical to that of the immunoprecipitated one. Therefore, the 15 protein subunits represent integral components of the Euglena oxidase. In an in vitro protein-synthesizing system using isolated mitochondria, polypeptides 1-3 were radioactive labeled in the presence of [35S]methionine. This further identifies these polypeptides with the three largest subunits of cytochrome c oxidase encoded by mitochondrial DNA in other eukaryotic organisms. By subtraction, the other 12 subunits can be assigned to nuclear genes. The isolated Euglena oxidase was highly active with Euglena cytochrome c558 and has monophasic kinetics. Using horse cytochrome c550 as a substrate, activity of the isolated oxidase was rather low. These findings correlate with the oxidase activity of mitochondrial membranes. Again, reactivity was low with cytochrome c550 and 35-fold higher with the Euglena cytochrome c558. The data show that the cytochrome c oxidase of the protist Euglena is different from other eukaryotic cytochrome c oxidases in number and size of subunits, and also with regard to kinetic properties and substrate specificity. PMID- 2545671 TI - Inhibition of the phosphate-stimulated cytochrome c oxidase activity by thiophosphate. AB - Yeast and mammalian cytochrome c oxidase activity is inhibited by thiophosphate. This inhibition was observed when using either whole mitochondria or the isolated or reconstituted enzyme. The kinetics of the reduction reaction enabled us to demonstrate that thiophosphate acted on the electron transfer between hemes a and a3. With whole mitochondria, phosphate alone stimulated respiration. The inhibition induced by thiophosphate was suppressed by phosphate only in mitochondria, but not when the isolated enzyme was used. The possibility of a kinetic regulation is discussed. PMID- 2545672 TI - Effect of cetyltrimethylammonium on ATP hydrolysis and proton translocation in the F0-F1 H+-ATP synthase of mitochondria. AB - The amphiphylic alkyl cation cetyltrimethylammonium inhibits the catalytic activity of soluble and membrane-bound F1 in a noncompetitive fashion. In sonic submitochondrial particles the Dixon plot showed a peculiar pattern with upward deviation at cetyltrimethylammonium concentration higher than 80 microM. In membrane-bound F1 the inhibition by cetyltrimethylammonium was potentiated by the F0 inhibitor ologomycin. Cetyltrimethylammonium also inhibited the oligomycin sensitive proton conductivity in F1-containing particles but was without any effect in F1-depleted particles. Also this inhibitory effect was potentiated by oligomycin. These results indicate functional cooperative interactions between F0 and F1. PMID- 2545673 TI - Overproduction and preliminary crystallographic study of ribonuclease H from Escherichia coli. AB - To facilitate the preparation of ribonuclease H from Escherichia coli in an amount sufficient for crystallographic studies, we have constructed an overproduction system for the enzyme. The structural gene for the enzyme was subcloned from pSK750 (Kanaya, S., and Crouch, R. J. (1983) J. Biol. Chem. 258, 1276-1281) to make a plasmid vector pPL801, in which the gene was under the control of bacteriophage lambda PL promoter. Thermal induction of the gene accumulated the enzyme in E. coli N4830-1 to approximately 8% of the total cytosolic protein. The level of production of the enzyme in N4830-1 harboring pPL801 was 14 mg/liter culture, which was 3000 times as high as that in the host cell. The enzyme was purified with a yield of more than 80% and crystallized by utilizing the property that the solubility of the enzyme decreased at pH values close to its isoelectric point (pI = 9). Crystals were grown by successive seeding (hanging drop method) for x-ray crystallographic analysis. The crystals belong to space group P212121 with unit cell dimensions of alpha = 44.1 A, b = 87.0 A, c = 35.5 A and contain one molecule in an asymmetric unit. They diffracted x-rays beyond 2.5 A resolution. PMID- 2545674 TI - Temporal integration of alpha 1-adrenergic responses in BC3H-1 muscle cells. Regulation of glycogen phosphorylase activity. AB - Regulation of Ca2+-dependent glycogen phosphorylase activity by alpha 1 adrenergic and H1-histamine receptors has been examined in BC3H-1 muscle cells. Stimulation by either norepinephrine or histamine elevates the phosphorylase activity ratio within 5 s from a resting value of 0.37 +/- 0.03 to maximal values of 0.8-0.9. Phosphorylase activation by alpha-adrenergic agonists is sustained over 20-30 min of agonist exposure, whereas histamine exposure only transiently activates phosphorylase during the initial 5 min of stimulation. The initial activation of phosphorylase by either receptor is not attenuated by treated cells with Ca2+-deficient and [ethylenebis(oxyethylenenitrilo)]tetraacetic acid supplemented buffer, whereas the response to sustained adrenergic stimulation depends largely, but not totally, upon extracellular Ca2+. The involvement of protein kinase C in agonist responses was tested by treating cells with phorbol 12-myristate 13-acetate. Phorbol 12-myristate 13-acetate inhibits receptor mediated mobilization of intracellular Ca2+ (IC50 = 3.6 nM) yet activates phosphorylase independently of agonist. Phorbol 12-myristate 13-acetate has no effect on cellular 45Ca2+ fluxes in the absence of agonist. Thus, the two receptors coordinately regulate intracellular signaling through Ca2+- and protein kinase C-mediated pathways. alpha 1-Adrenergic receptors elicit sustained phosphorylase activation whereas H1-histaminergic receptors desensitize. PMID- 2545675 TI - Regulation of the expression of the L-type pyruvate kinase gene in adult rat hepatocytes in primary culture. AB - Hepatocytes isolated from adult fasted rats and cultured in the presence of thyroid hormones, glucocorticoids, and in a serum-free medium conserve the essentials of their differentiated function and hormonal sensitivity for at least 1 week. In these cells, the gene for L-type pyruvate kinase is expressed only when glucose and insulin are present together, each of them being inactive by itself. Inhibition of the expression of the L-type pyruvate kinase gene which occurs when glucose and/or insulin are removed from the culture medium is not associated with accumulation of the phosphoenolpyruvate carboxykinase mRNA, which argues against the involvement of intracellular cyclic AMP in this phenomenon. Rather, a transcriptional activator, derived from carbohydrate metabolism and accumulating in the presence of insulin, seems to be needed to support the expression of the L-type pyruvate kinase gene. Glucagon, in vitro as in vivo, inhibits production of the L-type pyruvate kinase mRNAs. In addition to their roles on the production of these mRNAs, glucose and insulin on the one hand and glucagon on the other have profound effects on the stability of the L-type pyruvate kinase messengers: the half-life of the mRNA whose production has been blocked by actinomycin D is 1 h in the presence of glucagon and 24 h in the presence of glucose and insulin. Glucagon and glucose/insulin partially antagonize each other's effect on mRNA stability. PMID- 2545676 TI - Tumor necrosis factor alpha inhibits gonadotropin hormonal action in nontransformed ovarian granulosa cells. A modulatory noncytotoxic property. AB - It has been suggested that resident ovarian macrophages may play a role in the regulation of ovarian function through local paracrine secretion of regulatory molecule(s). It is the objective of the in vitro studies reported herein to evaluate the potential ovarian relevance of one such macrophage product, tumor necrosis factor alpha (TNF-alpha). To this end, use was made of a primary culture system of rat ovarian granulosa cells, the functional status of which was monitored by the acquisition of estrogen, progestin, and proteoglycan biosynthetic capacity. Whereas treatment with the gonadotropin follicle stimulating hormone (FSH), a potent functional regulator, resulted in a substantial increase in the extent of aromatization (conversion of androgenic steroid precursors to estrogens), concomitant exposure to TNF-alpha (10 ng/ml) produced significant (p less than 0.05), yet reversible inhibition (71 +/- 7%) of this FSH effect. This specific activity of TNF-alpha was characterized by a projected minimal effective dose of less than 0.1 ng/ml, an apparent median inhibitory dose of 0.56 +/- 0.14 ng/ml, and a minimal time requirement of 48 h. Significantly, the direct effect of TNF-alpha could not be accounted for by a decrease in cellular viability or plating efficiency, nor by a decrease in the number of cells or their DNA content. Instead, TNF-alpha inhibited FSH hormonal action at the level of stimulatable adenylate cyclase activity, exerting no apparent effect either at the level of the FSH receptor or at site(s) of action distal to cAMP generation. The effect of TNF-alpha was not limited to the attenuation of estrogen biosynthesis, exerting qualitatively similar effects on FSH-supported progestin and proteoglycan biosynthetic capacity. As such, these findings are in keeping with the notion that subnanomolar concentrations of TNF alpha, possibly of ovarian macrophage origin, may comprise the signal of a paracrine loop designed to attenuate gonadotropin action thereby playing a potential role in the development and/or demise of the ovarian follicle. PMID- 2545677 TI - A partial methylation profile for a CpG site is stably maintained in mammalian tissues and cultured cell lines. AB - We wished to determine if a partial methylation profile for a specific CpG site was stably maintained in both mammalian tissues and cultured cell lines. To accomplish this, we identified a CpG site with a partial methylation profile located upstream of the mouse adenine phosphoribosyltransferase promoter region. This site was found to be methylated at a level of approximately 25% in mouse brain, kidney, lung, and skeletal muscle tissues, at a level close to 50% in liver, and at level close to 0% in testis. These tissue-specific methylation profiles were not altered during aging. A methylation profile of approximately 25% at this CpG site was also observed in five mouse teratocarcinoma stem cell lines and one additional cultured cell line. This profile, however, was altered upon cellular differentiation, adenine phosphoribosyltransferase hemizygosity, and a loss of adenine phosphoribosyltransferase activity in some of the cultured cell lines. We conclude that partial methylation of a specific CpG site can be stably maintained both in vivo and in vitro and that a mechanism exists for its maintenance. The functional significance of a partial methylation profile remains to be determined. PMID- 2545678 TI - Topology of the transposon Tn10-encoded tetracycline resistance protein within the inner membrane of Escherichia coli. AB - The transposon Tn10-encoded tetracycline resistance protein TetA is an integral membrane protein responsible for the export of tetracycline from the cytoplasmic to the periplasmic side of the inner membrane of Gram-negative bacteria. From a plot of the average hydrophobicity along the sequence of this protein, a two dimensional membrane topology with 12 transmembrane domains may be predicted. Using plasmid-bearing Escherichia coli maxicells we specifically radiolabeled the TetA protein. The amino terminus of this membrane protein was shown not to be processed, and its location on the inner side of the cytoplasmic membrane was demonstrated by a newly developed use of a chemical method. Spheroplasts and inside-out vesicles of the TetA protein synthesizing maxicells were subjected to limited digestion by proteases of different specificities. The TetA protein was not accessible to proteases from the periplasmic side. On the inner side of the cytoplasmic membrane, the carboxyl terminus and four sites accessible to endoproteases could be identified. The cleavage sites are proposed to be localized between amino acid residues 60-70, 110-130, 180-200, and at amino acid 327. These results allow the definition of a model for the two-dimensional topology of the TetA protein. PMID- 2545679 TI - Interaction of the gamma-subunit of retinal rod outer segment phosphodiesterase with transducin. Use of synthetic peptides as functional probes. AB - There is considerable evidence which suggests that the gamma-subunit of cGMP phosphodiesterase (PDE gamma) is a multifunctional protein which may interact directly with both the catalytic subunits of PDE (PDE alpha beta) and the alpha subunit of transducin (T alpha) (Whalen, M., and Bitensky, M. (1989) Biochem. J. 259, 13-19; Griswold-Prenner, I., Young, J. H., Yamane, H. K., and Fung, B. K.-K. (1988) Invest. Ophthalmol. & Visual Sci. 29, (Suppl.) 218). To determine the region of interaction between the multifunctional PDE gamma and T alpha, and to determine the significance of this interaction, peptides corresponding to various regions of PDE gamma were synthesized and tested for their ability to inhibit the GTPase activity of T alpha. One of these peptides, PDE gamma-3 (bovine amino acid residues 31-45), inhibited the GTPase activity of T alpha with an I50 of 450 microM. The peptide (PDE gamma-3) was found to inhibit the GTPase activity of T alpha by inducing the binding of transducin to the rod outer segment membrane and by altering the GTP/GDP exchange. Analogs of PDE gamma-3 were synthesized to determine the required structure of the PDE gamma-3 region needed for the interaction of PDE gamma with T alpha. The results of these studies indicated that the removal of the positively charged amino acids or any of the potential hydrogen-bonding amino acids increased the I50 for the inhibition of the GTPase activity of T alpha Substitution of the hydrophobic amino acids had no effect. These results indicate the hydrophilic interactions may be essential for the binding of PDE gamma to T alpha and for the inhibition of the GTPase activity of T alpha by PDE gamma. The observed effects of PDE gamma-3 on T alpha and on PDE suggest that PDE gamma is a multifunctional protein which may play more than one role in the deactivation of the retinal transduction cascade. PMID- 2545680 TI - Ligand-induced dimerization of the platelet-derived growth factor receptor. Monomer-dimer interconversion occurs independent of receptor phosphorylation. AB - The platelet-derived growth factor (PDGF) receptor is a single membrane-spanning polypeptide of 180,000 daltons with a ligand-stimulatable tyrosine kinase site. We have investigated changes in the structure and association state of the receptor that are induced by ligand binding, but which precede autophosphorylation. Chemical cross-linking of PDGF-bound 32P-labeled receptor and 125I-PDGF-labeled receptor resulted in the generation of a radiolabeled cross linked complex of 370-390 kDa. This band, as well as the 180-190-kDa PDGF receptor band, were recognized by a PDGF receptor-specific antipeptide antibody. The appearance of the 370-390-kDa band was PDGF-dependent and was seen irrespective of whether the receptor was membrane-bound, solubilized, or highly (approximately 90%) purified. Sedimentation analysis of the 125I-PDGF cross linked receptor showed that both 180-190- and 370-390-kDa labeled species sedimented as a single peak at about 11.5 S, a position expected of a receptor dimer, demonstrating that the liganded receptor exists essentially as a dimer. In contrast, unliganded receptors sedimented as a single species at 7 S, a position consistent with a monomeric structure. The monomer-dimer interconversion was absolutely ligand-dependent and occurred independent of autophosphorylation. These results demonstrate and intimate correlation between PDGF binding and inter receptor bond formation, and raise the possibility that the phenomenon may be causally linked to the process of kinase activation. PMID- 2545681 TI - Identification, purification, and characterization of truncated forms of the human nerve growth factor receptor. AB - We report the presence of truncated forms of the nerve growth factor receptor (NGFRt) in the conditioned medium of the human melanoma cell line A875 and in human urine and amniotic fluid. Radioiodinated nerve growth factor (125I-NGF) specifically bound to NGFRt was chemically cross-linked. After immunoprecipitation, labeled receptor species were visualized by autoradiography following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. NGFRts were purified from human adult male urine or a mixture of human amniotic fluid and infant urine by using a combination of either ion exchange chromatography (adult) or ammonium sulfate precipitation (infant) and immunoaffinity chromatography. Typical yields were about 1 microgram/liter of adult urine and 75 micrograms/liter of amniotic fluid/infant urine. The purified proteins, with molecular masses of 45, 40, and 35 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (12%), were confirmed to be NGFRts by amino-terminal sequencing and were designated NGFRt-1, NGFRt-2, and NGFRt-3, respectively. The isoelectric points of these three species ranged from 3.3 to 3.95 and displayed intraspecies heterogeneity; subsequently, amino acid residues covalently modified with sialic acid-containing carbohydrates were documented. The binding affinities of these species for nerve growth factor were comparable to that of the low affinity cell surface receptor. The potential to isolate milligram quantities of human NGFRts allows for model studies of the physicochemical structure of the intact receptor and the generation of polyclonal antibodies to study the biological functions of the NGF receptor. PMID- 2545682 TI - Kinetic analysis of the formation of inositol 1:2-cyclic phosphate in carbachol stimulated pancreatic minilobules. Half is formed by direct phosphodiesteratic cleavage of phosphatidylinositol. AB - The issue as to whether there is direct phosphodiesteratic cleavage of phosphatidylinositol (PI), in addition to that of phosphatidylinositol 4,5 bisphosphate (PIP2), on agonist stimulation of cells has been controversial. In an attempt to resolve this issue, we have studied the kinetics of the formation and breakdown of the cyclic inositol phosphates. This approach is fairly straightforward, since the turnover of the cyclic inositol phosphates is very slow as compared to that of the noncyclic inositol phosphates and proceeds from inositol 1:2-cyclic 4,5-trisphosphate to inositol 1:2-cyclic phosphate (I(c1:2)P) directly by dephosphorylation without any branching pathways, in contrast to the multiple branchpoints of the noncyclic inositol phosphate pathway. Mouse pancreatic minilobules were prelabeled with [3H]inositol for 30 min, followed by washing to remove free inositol. They were then stimulated with carbachol for 30 min. The inositol cyclic polyphosphates reached steady state at 10-15 min, and I(c1:2)P reached steady state at 25 min. We blocked the action of carbachol by addition of an excess of atropine at 30 min, and the rate of disappearance of the three cyclic inositol phosphates was measured. From these data, the contribution of the inositol cyclic polyphosphate pathway to I(c1:2)P was calculated, which was 40-50% of total I(c1:2)P formation. Thus, 40-50% of the I(c1:2)P formed must have been derived from direct phosphodiesteratic cleavage of PI. This approach should prove useful in measuring the relative contributions of PI hydrolysis and PI phosphorylation (phosphatidylinositol 4,5-bisphosphate hydrolysis) in the overall PI cascade. PMID- 2545684 TI - Examination of the function of active site lysine 329 of ribulose-bisphosphate carboxylase/oxygenase as revealed by the proton exchange reaction. AB - Diverse approaches that include site-directed mutagenesis have indicated a catalytic role of Lys-329 of ribulosebisphosphate carboxylase/oxygenase from Rhodospirillum rubrum. To determine whether Lys-329 is required for the initial enolization of ribulose bisphosphate or for some subsequent step in the overall reaction pathway, the competence of position 329 mutant proteins (devoid of carboxylase activity) in catalyzing exchange of solvent protons with the C-3 proton of substrate has now been examined. Irrespective of the amino acid substitution for Lys-329, the mutant protein retains 2-6% of the wild-type activity in the proton exchange reaction. The complete stability of ribulose bisphosphate during the enolization catalyzed by mutant protein suggests that the major effect of Lys-329 is to facilitate the addition of gaseous substrates (CO2 or O2) to the enediol intermediate. The exchange reaction requires Mg2+, is CO2 dependent, and is inhibited by the transition-state analogue 2-carboxyarabinitol 1,5-bisphosphate. A mutant protein in which Lys-191, the site for carbamylation by CO2 in an obligatory activation step, is replaced by a cysteinyl residue totally lacks proton exchange activity. Barely detectable exchange activity (approximately 0.2% of wild-type) is displayed by the Lys-166----Cys mutant protein, consistent with the previously implicated role of Lys-166 in the deprotonation of ribulose bisphosphate. Retention of exchange activity by the Glu 48----Gln mutant protein, which is slightly active in overall carboxylation, demonstrates that active site Glu-48, like Lys-329, exerts its major effect at some step subsequent to the initial enolization. PMID- 2545683 TI - The "regulatory" sulfhydryl group of Penicillium chrysogenum ATP sulfurylase. Cooperative ligand binding after SH modification; chemical and thermodynamic properties. AB - ATP sulfurylase from Penicillium chrysogenum is a homohexamer that contains three free sulfhydryl groups/subunit, only one of which (designated SH-1) can be modified by disulfide, maleimide, and halide reagents under nondenaturing conditions. Modification of SH-1 has only a small effect on kcat but causes the [S]0.5 values for MgATP and SO4(2-) (or MoO4(2-) to increase by an order of magnitude. Additionally, the velocity curves become sigmoidal with a Hill coefficient (nH) of about 2 (Renosto, F., Martin, R. L., and Segel, I. H. (1987) J. Biol. Chem. 262, 16279-16288). Direct equilibrium binding measurements confirmed that [32P]MgATP binds to the SH-modified enzyme in a positively cooperative fashion (nH = 2.0) if a sulfate subsite ligand (e.g. FSO3-) is also present. [35S]Adenosine 5'-phosphosulfate (APS) binding to the SH-modified enzyme displayed positive cooperativity (nH = 1.9) in the absence of a PPi subsite ligand. The results indicate that positive cooperativity requires occupancy of the adenylyl and sulfate (but not the pyrophosphate) subsites. [35S]APS binding to the native enzyme displayed negative cooperativity (or binding to at least two classes of sites). Isotope trapping profiles for the single turnover of [35S]APS: (a) confirmed the equilibrium binding curves, (b) indicated that all six sites/hexamer are catalytically active, and (c) showed that APS does not dissociate at a significant rate from E.APS.PPi. The MgPPi concentration dependence of [35S]APS trapping was indicative of MgPPi binding to two classes of sites on both the native and SH-modified enzyme. Inactivation of the native or SH modified enzyme by phenylglyoxal in the presence of saturating APS was biphasic. The semilog plots suggested that only half of the sites were highly protected. The cumulative data suggest a model in which pairs of sites or subunits can exist in three different states designated HH (both sites have a high APS affinity, as in the native free enzyme), LL (both sites have a low APS affinity as in the SH modified enzyme), and LH (as in the APS-occupied native or SH-modified enzyme). Thus, the HH----LH transition displays negative cooperativity for APS binding while the LL----LH transition displays positive cooperativity. The relative reactivities of like-paired SH-reactive reagents were in the order: N phenylmaleimide greater than N-ethylmaleimide; dithionitropyridine greater than dithionitrobenzoate; thiolyte-MQ greater than thiolyte-MB. The log kmod versus pH curve indicates that the pKa of SH-1 is greater than 9.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2545686 TI - Ha-ras activates the Na+/H+ antiporter by a protein kinase C-independent mechanism. AB - In quiescent Ha-ras-transfected NIH 3T3 cells, addition of serum growth factors, bombesin or 12-O-tetradecanoylphorbol-13-acetate (TPA) leads to a dimethylamiloride-sensitive intracellular alkalinization which can be inhibited by staurosporine, a potent inhibitor of protein kinase C. Expression of the transforming Ha-ras gene causes a growth factor-independent increase in cytoplasmic pH. This Ha-ras-induced alkalinization is sensitive to dimethylamiloride but is not affected by staurosporine concentrations which prevent the pH response after addition of growth factors or TPA. Protein kinase C depletion by long term exposure to TPA eliminates the pH response to bombesin and phorbol ester but does not effect the Ha-ras-induced intracellular alkalinization. It is concluded that expression of Ha-ras causes an activation of the Na+/H+ antiporter by an as yet unknown protein kinase C-independent mechanism. PMID- 2545685 TI - Studies on the active site of the Neurospora crassa plasma membrane H+-ATPase with periodate-oxidized nucleotides. AB - The Neurospora crassa plasma membrane H+-ATPase is inactivated by the periodate oxidized nucleotides, oATP, oADP, and oAMP, with oAMP the most effective. Inhibition of the ATPase is essentially irreversible, because Sephadex G-50 column chromatography of the oAMP-treated ATPase does not result in a reversal of the inhibition. Inhibition of the ATPase by oAMP is protected against by the H+ ATPase substrate ATP, the product ADP, and the competitive inhibitors TNP (2',3' O-(2,4,6-trinitrocyclohexadienylidine)-ATP and TNP-ADP, suggesting that oAMP inhibition occurs at the nucleotide binding site of the enzyme. The rate of inactivation of the ATPase by oAMP is only slightly affected by EDTA, indicating that the oAMP interaction with the nucleotide binding site of the H+-ATPase occurs in the absence of a divalent cation. The protection against oAMP inhibition by ADP is likewise unaffected by EDTA. The inhibition of the ATPase by oAMP is absolutely dependent on the presence of acidic phospholipids or acidic lysophospholipids known to be required for H+-ATPase activity, suggesting that these lipids either aid in the formation of the nucleotide binding site or render it accessible. Incubation of the ATPase with Mg2+ plus vanadate, which locks the enzyme in a conformation resembling the transition state of the enzyme dephosphorylation reaction, completely protects against inhibition by oAMP, suggesting that in this transition state conformation the nucleotide site either does not exist, or is inaccessible to oAMP. Labeling studies with [14C] oAMP indicate that the incorporation of 1 mol of oAMP is sufficient to cause complete inactivation of the ATPase. PMID- 2545687 TI - Characterization of an imidazoline/guanidinium receptive site distinct from the alpha 2-adrenergic receptor. AB - alpha 2-Adrenergic receptors recognize a number of molecules with diverse chemical structures, including the yohimban diastereoisomers yohimbine and rauwolscine, catecholamines, guanidinium analogs, and imidazolines, such as clonidine. The affinity of the receptor protein for some of these ligands can vary by 10-100-fold among various tissues and species, suggesting a heterogeneous class of binding sites. Certain cellular effects elicited by the compounds possessing an imidazoline or guanidinium moiety may actually be mediated by a membrane receptor distinct from the alpha 2-adrenergic receptor. To determine whether this imidazoline/guanidinium receptive site (IGRS) and the alpha 2 adrenergic receptor represent distinct proteins, we solubilized and partially characterized the two binding sites in rabbit kidney. This tissue expresses both alpha 2-adrenergic receptors and high affinity imidazoline/guanidinium binding sites, the latter which are rauwolscine-insensitive but can be identified with the benzodioxan [3H]idazoxan. The IGRS and alpha 2-adrenergic receptor in rabbit kidney exhibit distinct ligand recognition properties, which are maintained after solubilization and partial purification. In addition, the two receptors can be physically separated by heparin-agarose or lectin affinity chromatography indicating that the two binding sites are distinct entities. [3H]Idazoxan binding is trypsin-sensitive, indicating that the IGRS is a protein rather than a lipid component of the plasma membrane. [3H]Idazoxan binding is not inhibited by endogenous agonists for known neurotransmitter receptors. However, the IGRS does recognize clonidine-displacing substance, a small non-catechol compound isolated from calf brain, suggesting the existence of a previously uncharacterized hormonal/neurotransmitter receptor system. PMID- 2545689 TI - Studies of crystalline trimethylamine dehydrogenase in three oxidation states and in the presence of substrate and inhibitor. AB - Crystals of trimethylamine dehydrogenase have been examined by difference Fourier methods at 6.0-A resolution after partial reduction by substrate and by dithionite in the presence of inhibitor. Similar studies of the inhibited oxidized enzyme and of the enzyme reduced fully by dithionite alone were also carried out. In all cases ligand binding at the active site occurred. In addition, there were small structural changes, possibly side chain movements, in the inhibited oxidized enzyme and somewhat larger changes in the partially reduced crystals. The largest changes occurred with the fully reduced enzyme. However, in no cases were subunit or domain movements observed nor were changes observed in the position of the FMN or [4Fe-4S] cofactors. Parallel studies of crystalline trimethylamine dehydrogenase were carried out by EPR spectroscopy. The results show that the electronic states of the crystalline enzyme under the conditions of the difference Fourier studies are comparable to those which occur in solution under similar conditions. PMID- 2545688 TI - Localization of 1,25-(OH)2D3-responsive alkaline phosphatase in osteoblast-like cells (ROS 17/2.8, MG 63, and MC 3T3) and growth cartilage cells in culture. AB - Previous studies have shown 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3)-responsive alkaline phosphatase in cultured growth zone cartilage chondrocytes is localized in extracellular matrix vesicles (MV). Since osteoblast-like cells also have 1,25 (OH)2D3-responsive alkaline phosphatase, this study determined whether the 1,25 (OH)2D3-responsive enzyme activity is localized to MV produced by these cells as well. Osteoblast-like cells from rat (ROS 17/2.8), mouse (MC 3T3), human (MG 63), and rat growth zone cartilage were cultured in Dulbecco's modified Eagle's medium containing 10(-7)-10(-12) M 1,25-(OH)2D3. Alkaline phosphatase total activity and specific activity were measured in the cell layer, MV, and plasma membrane (PM) fractions. MV and PM purity were verified by electron microscopy and MV alkaline phosphatase specific activity compared to PM (MV versus PM: ROS 17/2.8 6 x; MG 63, 5.5 x; MC 3T3, 33 x; GC, 2 x). There was a dose-dependent stimulation of MV alkaline phosphatase (5- to 15-fold increase at 10(-7)-10(-9) M) in all cell types in response to the 1,25-(OH)2D3. The PM enzyme was stimulated in a parallel fashion in the osteoblast cultures. No effect of 1,25-(OH)2D3 was observed in growth cartilage PM. Although MV accounted for less than 20% of the total activity they contributed 50% of the increase in alkaline phosphatase activity in the cell layer in response to 1,25-(OH)2D3 and MV specific activity was enriched 10 times over that of the cell layer. These are common features of MV produced by cells which calcify their matrix and suggest that hormonal regulation of MV enzymes may be important in primary calcification. PMID- 2545690 TI - The role of phosphorylation and limited proteolytic cleavage of talin and vinculin in the disruption of focal adhesion integrity. AB - Chemical agents which activate specific kinases were employed to disrupt the stress fiber and focal adhesion organization of cells spread on a substratum. The phorbol ester 12-O-tetradecanoylphorbol-13-acetate, an activator of protein kinase C, promoted a rapid loss of stress fibers and focal adhesions from African green monkey kidney (BSC-1) cells. This was paralleled by an increase in the level of talin phosphorylation suggesting that this may play a role in the removal of talin from focal adhesions. Similar morphological changes were produced in the rat embryo fibroblast line (REF 52) by dibutyryl-cAMP, which stimulates protein kinase A. In contrast, however, the phosphorylation of talin was reduced in REF 52 cells when treated with dibutyryl cAMP. In untreated cells we found that the levels of vinculin phosphorylation were very low relative to the levels of talin phosphorylation and did not change following drug treatment in either cell line. Although limited proteolytic cleavage of cytoskeletal proteins represents a potential mechanism for focal adhesion disruption, we observed no proteolysis of talin or vinculin in response to either drug treatment. PMID- 2545691 TI - Mouse angiotensin-converting enzyme is a protein composed of two homologous domains. AB - Angiotensin-converting enzyme (ACE) is a dipeptidyl carboxypeptidase that converts angiotensin I into the potent vasoconstrictor angiotensin II. We have used cDNA and genomic sequences to assemble a composite cDNA, ACE.315, encoding the entire amino acid sequence of mouse converting enzyme. ACE.315 contains 4838 base pairs and encodes a protein of 1278 amino acids (147.4 kDa) after removal of a 34-amino acid signal peptide. Within the protein, there are two large areas of homologous sequence, each containing a potential Zn-binding region and catalytic site. These homologous regions are approximately half the size of the whole ACE protein and suggest that the modern ACE gene is the duplicated product of a precursor gene. Mouse ACE is 83% homologous to human ACE in both nucleic acid and amino acid sequence, and like human ACE, contains a hydrophobic region in the carboxyl terminus that probably anchors the enzyme to the cell membrane (Soubrier, F., Alhenc-Gelas, F., Hubert, C., Allegrini, J., John, M., Tregear, G., and Corvol, P. (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 9386-9390). Northern analysis of mouse kidney, lung, and testis RNA demonstrates that the testicular isozyme of ACE is encoded by a single, smaller RNA (2500 bases) than the two message sizes found in kidney or lung (4900 and 4150 bases), and that this testicular RNA hybridizes to the 3' portion of ACE.315. PMID- 2545692 TI - Purification and biologic characterization of a specific tumor necrosis factor alpha inhibitor. AB - The purpose of the present investigation was to purify a urine-derived tumor necrosis factor alpha inhibitor (TNF alpha INH) and to characterize its mechanism of action. For the purification procedure, urine was concentrated and TNF alpha INH purified by ion-exchange chromatographies, gel filtration, TNF alpha affinity column, and reverse-phase chromatography. The TNF alpha INH migrates with an apparent Mr of approximately 33,000 when estimated on sodium dodecyl sulfate polyacrylamide gel electrophoresis run under both reducing and nonreducing conditions. Elution of TNF alpha INH activity from the gel yields also a approximately 33,000-Da inhibitory fraction. Besides inhibiting TNF alpha-induced cytotoxicity in L929 cells in the presence of actinomycin D, the TNF alpha INH impeded in a dose-dependent manner prostaglandin E2 production and expression of cell-associated interleukin-1 by human dermal fibroblasts. Therefore, TNF alpha INH is active on both actinomycin D-treated and untreated cells. In contrast to TNF alpha, TNF beta-induced cytotoxicity was only slightly affected by the inhibitor. This specificity was confirmed by the fact that it affected neither interleukin-1 alpha nor interleukin-1 beta biologic activities. The mechanism of action of TNF alpha INH involves blocking of 125I-TNF alpha binding to the promonocytic cell line U937. Moreover, preincubation of 125I-TNF alpha with TNF alpha INH increased binding inhibition, suggesting an interaction between TNF alpha and the inhibitor. PMID- 2545693 TI - A tumor necrosis factor-binding protein purified to homogeneity from human urine protects cells from tumor necrosis factor toxicity. AB - Unfractionated preparations of the proteins of human urine provided protection against the in vitro cytocidal effect of tumor necrosis factor (TNF). In certain cells, the proteins decreased expression of the receptors for TNF in a temperature-dependent way. In all cells examined, the proteins were found to interfere also with the binding of both TNF and interleukin-1 when applied directly into the binding assays. That effect could be observed in the cold, suggesting that it was independent of cellular metabolism. A protein which protects cells against the cytotoxicity of TNF was purified from human urine by chromatography on CM-Sepharose followed by high performance liquid chromatography on Mono Q and Mono S columns and reversed phase high performance liquid chromatography. This protein is a very minor constituent of normal urine, with an apparent molecular weight of about 27,000 in sodium dodecyl sulfate polyacrylamide gel electrophoresis under both reducing and nonreducing conditions. Homogeneity of the purified protein was confirmed by microsequence analysis which revealed a single N-terminal sequence: Asp-Ser-Val-Cys-Pro-. The protein protected cells from TNF toxicity at concentrations of a few nanograms per ml and interfered with the binding of both TNF-alpha and TNF-beta to cells, when applied simultaneously with the cytokines. However, unlike crude preparations of the urinary proteins, the purified protein did not induce in cells a decrease in ability to bind TNF nor did it interfere with the binding of interleukin-1 to its receptor. Direct, specific binding to the protein of TNF alpha and, to a lesser extent, also TNF-beta, but not of interleukin-1 nor interferon-gamma could be demonstrated. It is suggested that this protein blocks the function of TNF by competing for TNF with the TNF receptor and not by interacting with the target cell. PMID- 2545694 TI - The cellular pool of Na+ channels in the amphibian cell line A6 is not altered by mineralocorticoids. Analysis using a new photoactive amiloride analog in combination with anti-amiloride antibodies. AB - An amiloride-sensitive Na+ channel is found in the apical plasma membrane of high resistance, Na+ transporting epithelia. We have developed a method for the identification of this channel based on the use of a new high affinity photoreactive amiloride analog, 2'-methoxy-5'-nitrobenzamil (NMBA), and anti amiloride antibodies to identify photolabeled polypeptides. NMBA specifically labels the putative Na+ channel in bovine kidney microsomes. A 130-kDa polypeptide is detected on immunoblots with anti-amiloride antibodies. NMBA is a potent inhibitor of Na+ transport in the established amphibian kidney epithelial cell line A6, and specifically labels a 130-kDa polypeptide. We utilized both NMBA photolabeling and [3H]benzamil binding in order to examine the cellular pool of putative channels following hormonal regulation of Na+ transport. This pool is not significantly altered by the mineralocorticoid agonist aldosterone or antagonist spironolactone, despite a 3.8-fold difference in transepithelial Na+ transport. PMID- 2545695 TI - Hyperosmolarity inhibits galactosyl receptor-mediated but not fluid phase endocytosis in isolated rat hepatocytes. AB - We have investigated the effects of hyperosmolarity induced by sucrose on the fluid phase endocytosis of the fluorescent dye lucifer yellow CH (LY) and the endocytosis of 125I-asialo-orosomucoid (ASOR) by the galactosyl receptor system in isolated rat hepatocytes. Continuous uptake of LY by cells at 37 degrees C is biphasic, occurs for 3-4 h, and then plateaus. Permeabilized cells or crude membranes do not bind LY at 4 or 37 degrees C. Intact cells also do not accumulate LY at 4 degrees C. The rate and extent of LY accumulation are concentration- and energy-dependent, and internalized LY is released from permeabilized cells. Efflux of internalized LY from washed cells is also biphasic and occurs with halftimes of approximately 38 and 82 min. LY is taken up into vesicles throughout the cytoplasm and the perinuclear region with a distribution pattern typical of the endocytic pathway. LY, therefore, behaves as a fluid phase marker in hepatocytes. LY has no effect on the uptake of 125I-ASOR at 37 degrees C. The rate of LY uptake by cells in suspension is not affected for at least 30 min by up to 0.2 M sucrose. The rate of endocytosis of 125I-ASOR, however, is progressively inhibited by increasing the osmolality of the medium with sucrose (greater than 98% with 0.2 M sucrose; Oka and Weigel (1988) J. Cell. Biochem. 36, 169-183). Hyperosmolarity completely inhibits endocytosis of 125I-ASOR by the galactosyl receptor, whereas fluid phase endocytosis of LY is unaffected. Cultured hepatocytes contained about 100 coated pits/mm of apical membrane length as assessed by transmission electron microscopy. In the presence of 0.4 M sucrose, only 17 coated pits/mm of membrane were observed, an 83% decrease. Only a few percent of the total cellular fluid phase uptake in hepatocytes is due to the coated pit endocytic pathway. We conclude that the fluid phase and receptor mediated endocytic processes must operate via two separate pathways. PMID- 2545696 TI - Transforming growth factor-beta (TGF-beta) receptor proteoglycan. Cell surface expression and ligand binding in the absence of glycosaminoglycan chains. AB - The type III transforming growth factor-beta (TGF-beta) receptor is a cell surface chondroitin/heparan sulfate proteoglycan that binds various forms of TGF beta with high affinity and specificity. Here, we have used a genetic approach to determine the requirement for glycosaminoglycan (GAG) chains for normal TGF-beta receptor expression and the role that the receptor proteoglycan core and GAG chains play in TGF-beta binding. Chinese hamster ovary (CHO) cells defective in GAG synthesis express on their surface 110-130-kDa type III receptor proteoglycan cores that can bind normal levels of TGF-beta compared to wild type CHO cells. The affinity of the receptor core for TGF-beta 1 and TGF-beta 2 in CHO cell mutants is similar to that of the TGF-beta receptor proteoglycan forms present in wild type CHO cells or in CHO cell mutants that have been allowed to bypass their metabolic defect and express the wild type proteoglycan phenotype. The binding properties of TGF-beta receptor types I and II in CHO cells and the growth inhibitory response of CHO cell mutants to TGF-beta are not impaired by the absence of GAG chains in the type III receptor. These results show that the GAG chains are dispensable for type III receptor expression on the cell surface, binding of TGF-beta to the receptor core, and growth inhibitory response of the cells to TGF-beta. The evidence also suggests that the type III receptor may act as a multifunctional proteoglycan able to bind TGF-beta via the receptor core while performing another as yet unidentified function(s) via the GAG chains. PMID- 2545698 TI - Mannose 6-phosphate receptors and lysosomal enzyme targeting. PMID- 2545697 TI - F0 "proton channel" of rat liver mitochondria. Rapid purification of a functional complex and a study of its interaction with the unique probe diethylstilbestrol. AB - The F0 portion of the rat liver mitochondrial ATP synthase (F0F1-ATPase) has been purified by a rapid, high yield procedure. F0 is selectively extracted from inner membrane vesicles with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) after prior treatment of the vesicles with guanidine HCl to remove F1. The resultant F0 is functional in proton translocation assays and separates in sodium dodecyl sulfate-polyacrylamide gel electrophoresis into four major and three minor Coomassie-stainable bands, all with apparent molecular masses below 30 kDa. This CHAPS-purified F0 preparation was characterized in detail for its capacity to interact with the unique probe diethylstilbestrol (DES) which, depending on conditions, has been shown to interact with rat liver F0F1 to either inhibit or promote ATP hydrolysis (McEnery, M. W., and Pedersen, P.L. (1986) J. Biol. Chem. 261, 1745-1752). DES-inhibitory sensitivity could be conferred on F1 ATPase activity with the same concentration dependence on F0 as conferral of oligomycin sensitivity. DES was shown also to inhibit the magnitude of valinomycin induced proton influx, while initiating proton efflux in asolectin vesicles reconstituted with F0 and loaded with K+. The potency of DES in producing the latter effects was shown to be highly dependent on hydroxyl groups in "para" positions of the two benzene rings within the DES molecule. Finally, in the absence of F0, DES was shown to act as a catalyst of proton influx in K+ loaded asolectin vesicles upon addition of valinomycin. A model based on the structure of DES is presented to account for both the inhibitory and uncoupling properties of this compound. PMID- 2545699 TI - A reactive cysteine in avian liver phosphoenolpyruvate carboxykinase. AB - The modification of avian phosphoenolpyruvate carboxykinase by a variety of sulfhydryl reagents leads to inhibition. The inhibition is related to the loss of 1 highly reactive cysteine residue of the 13 cysteines present in the enzyme. Inhibition by reagents which yield a mixed disulfide was rapidly reversed by thiols. Reagents specific for vicinal sulfhydryl configurations were not potent inhibitors. The cysteine-modified enzyme continues to bind Mn2+ with the same stoichiometry and dissociation constant as the native enzyme. All of the substrates also bind to thiol-modified inactive enzyme. The modification of the reactive cysteine with the spin-labeled iodoacetate derivative leads to inactive enzyme with spin label stoichiometrically incorporated. The EPR spectrum showed an immobilized spin label on the enzyme. EPR studies of the perturbation of the phosphoenolpyruvate carboxykinase-bound spin label by bound Mn2+ showed a dipolar interaction between the two spins, estimated to be 10 A apart. The perturbation of the 1/T1 and 1/T2 values of the 31P resonances of ITP by spin-labeled enzyme indicates that this portion of the nucleotide binds 8-10 A from the spin label. These results indicate that the reactive cysteine is close to but not at the active site of the enzyme. The thiol group must be free and in its reduced form for the enzyme to be active. Perhaps modification of this group prevents conformational change(s) upon ligand binding necessary for the catalytic process. PMID- 2545700 TI - Reduced activity of the interferon-induced double-stranded RNA-dependent protein kinase during a heat shock stress. AB - Previous studies have shown that the antiviral response induced by interferon in murine cells could be degraded after a heat shock. Here we have confirmed that a similar effect occurs also in interferon-treated human HeLa cells subjected to a heat shock. In addition, we have investigated the fate of the interferon-induced, double-stranded RNA-dependent protein kinase in heat-shocked cells. This protein kinase is a Mr 68,000 protein (p68 kinase) which, when autophosphorylated, catalyzes phosphorylation of the protein synthesis eukaryotic initiation factor 2, thus mediating inhibition of protein synthesis. After heat shock of interferon treated HeLa cells, the double-stranded RNA-dependent autophosphorylation of p68 kinase in cytoplasmic extracts is greatly reduced whereas the phosphorylation of other cellular proteins is not affected. In vivo, autophosphorylation of p68 kinase is also reduced in heat-shocked cells whereas there is no apparent effect on the phosphorylation state of other proteins. In such cells, the interferon mediated antiviral response becomes modified according to the virus challenge, i.e. these cells remain resistant to vesicular stomatitis virus but become partially sensitive to encephalomyocarditis virus (EMCV) infection. The reduction in the activity of p68 kinase is due to its reduced nonionic detergent solubility occurring during the heat shock period. The resultant reduced detergent extractibility of p68 kinase is dependent on the intensity of the thermal stress. In contrast to the effect after a heat shock, arsenite treatment of interferon treated HeLa cells induces heat shock proteins, but neither modifies the antiviral response nor affects the extractibility of p68 kinase. These results indicate that the degradation of the anti-EMCV response and reduced p68 kinase activity occur in response to heat treatment independently of the induction of heat shock proteins. The role of p68 kinase in the mechanism of the antiviral response against EMCV and vesicular stomatitis virus is discussed. PMID- 2545701 TI - Intracellular Mn (II)-associated superoxide scavenging activity protects Cu,Zn superoxide dismutase-deficient Saccharomyces cerevisiae against dioxygen stress. AB - Three Cu,Zn superoxide dismutase (SOD-1)-deficient Saccharomyces cerevisiae mutants do not grow in 100% O2 in rich medium and require Met and Lys when grown in air (Bilinski, T., Krawiec, Z., Liczmanski, A., and Litwinska, J. (1985) Biochem. Biophys. Res. Commun. 130, 533-539). We show herein that medium manganese (II) accumulated by the mutants rescues these O2-sensitive phenotypes; 2 mM medium Mn2+ represented the threshold required for cell growth. The accumulation of Mn2+ was not oxygen-inducible since mutants grown aerobically and anaerobically accumulated the same amount of Mn2+. Mn2+ accumulation is not unique to these mutants since wild type accumulated almost twice as much Mn2+ as did mutant. ESR spectra of the cell extracts and whole cells loaded with Mn2+ were typical of free Mn(II) ion. These spectra could not account quantitatively for the total cellular Mn2+, however. A screen for soluble antioxidant activities in the Mn2+-supplemented cells detected O2- (superoxide) scavenging activity, with no change in catalase or peroxidase activities. This O2- scavenging activity was CN- and heat-resistant. No achromatic bands were revealed in nondenaturing gels of Mn2+- containing cell extracts stained for O2- scavenging activity. The Mn2+-dependent O2- scavenging activity in the cell extracts was quenched by EDTA and dialyzable. More than 60% of both the intracellular Mn2+ and the O2- scavenging activity was removed by 2-h dialysis. Dialyzed cells were not viable in air unless resupplemented with either Met or Mn2+. Although Mn2+ supported the aerobic growth of these mutants, excess Mn2+, which correlated with an elevated O2- scavenging activity, was toxic to both mutant and wild type. The results indicate that free or loosely bound Mn2+ ion protects the mutants against oxygen stress by providing an intracellular, presumably cytosolic, O2- scavenging activity which replaces the absent SOD-1. PMID- 2545702 TI - A soluble form of bovine rod photoreceptor phosphodiesterase has a novel 15-kDa subunit. AB - A substantial fraction (20-30%) of the bovine rod outer segment phosphodiesterase (PDE) activity is not associated with outer segment membranes prepared with buffers of moderate ionic strength; this PDE activity appears to represent a distinct, soluble isozyme. Although this PDE isozyme can be demonstrated to be present in sealed rod outer segments, it is discarded from most standard rod outer segment preparations. A method was developed that allowed the rapid purification of the soluble rod PDE by 2600-fold, to apparent homogeneity, using a monoclonal antibody column (ROS-1a). The soluble rod PDE isozyme has a novel Mr = 15,000 subunit (delta) in addition to subunits of Mr = 88,000 (alpha sol), 84,000 (beta sol), and 11,000 (gamma sol). The delta subunit comigrates with and may be identical to the cone PDE 15-kDa subunit. The small subunits of the soluble rod PDE and the membrane-associated rod PDE were isolated by reverse phase chromatography. The gamma sol subunit was a potent inhibitor of trypsin activated rod PDE, inhibiting 50% of 1 pM PDE activity at a concentration of 11 pM. This concentration was similar to that observed for the gamma subunit of the membrane-associated rod PDE. The purified delta subunit did not appear to affect PDE activity; this subunit was, however, unusually difficult to keep in solution. All of the kinetic and physical properties of the soluble rod PDE tested thus far are similar to those of the membrane-associated form, except for the presence of the delta subunit, suggesting that this unique subunit could mediate the solubility of the soluble rod PDE and the cone PDE in the intact photoreceptor. PMID- 2545703 TI - The bacteriophage T4 DNA replication fork. Only DNA helicase is required for leading strand DNA synthesis by the DNA polymerase holoenzyme. AB - Seven bacteriophage T4-encoded proteins reconstitute a DNA replication apparatus that catalyzes coupled leading and lagging strand DNA synthesis at a replication fork in vitro. The proteins involved are the T4 DNA polymerase holoenzyme (the products of T4 genes 43, 44/62, and 45), a helix-destabilizing (SSB) protein (gene 32 protein), and the T4 primosome which is composed of a DNA helicase (gene 41 protein) and a primase (gene 61 protein). We show here that the presence of 41 protein on the lagging strand of the fork enables the polymerase holoenzyme to catalyze leading strand DNA synthesis at a maximum rate and with high processivity. This leading strand synthesis is unaffected by the addition of either the gene 32 or the gene 61 protein; the 41 protein cannot be replaced by the dda protein, a second T4-encoded DNA helicase. When the 61 protein is added to the 41 protein to complete the primosome, Okazaki fragment synthesis on the lagging strand accompanies leading strand DNA synthesis in this system even in the absence of the 32 protein. However, the addition of 32 protein decreases the size of the Okazaki fragments made, as expected for an increase in the lagging strand polymerization rate at a fork that has coupled leading and lagging strand DNA polymerase molecules. PMID- 2545704 TI - The encoded primary sequence of a rice seed ADP-glucose pyrophosphorylase subunit and its homology to the bacterial enzyme. AB - Rice seed ADP-glucose pyrophosphorylase cDNA clones were isolated by screening a lambda expression library prepared from rice endosperm poly(A+) RNA with a heterologous antibody raised against the spinach leaf enzyme and subsequently by nucleic acid hybridization. One cDNA plasmid, possessing about 1650 nucleotides, was shown by both DNA and RNA sequence analysis to contain the complete ADP glucose pyrophosphorylase coding sequence of 483 amino acids. The primary sequence displayed a putative leader peptide presumably required for transport of this nuclear encoded protein into the amyloplasts, a differentiated starch containing plastid. The leader peptide, however, showed little sequence homology with transit peptides displayed by other known nuclear encoded proteins localized in the chloroplasts. A comparison of the primary sequence of the putative mature subunit to the Escherichia coli pyrophosphorylase showed two regions displaying significant homology. These two conserved regions contain residues shown previously to be essential for the allosteric regulation and catalytic activity of the E. coli enzyme. Differences in the primary sequences of the plant and bacterial enzyme may reflect the distinct nature of the allosteric effectors that control these enzymes. PMID- 2545705 TI - Rapid rescue of cellular transcriptional activator elements by amplification of a single copy selection gene. AB - Cellular transcriptional activator sequences from a Syrian hamster cell line (baby hamster kidney (BHK] were rescued by a double selection procedure. An enhancer-deficient SV40 promoter was linked to the neomycin resistance (NEO) gene and transfected into BHK cells. Genomic DNA fragments of G418-resistant cell clones containing multiple copies of integrated plasmid DNAs were used for a second transfection of BHK cells, resulting in the genomic integration of a single copy plasmid which expresses the NEO gene efficiently. For rapid cloning of the integrated promoter and adjacent cellular DNA sequences, these cell clones were fused to COS-1 cells, thereby providing SV40 large T antigen and the monkey cell permissive factor necessary for SV40 replication. Resulting from this fusion, the integrated plasmid and adjacent sequences were amplified to about 1000 extrachromosomal copies giving rise to an abundant pool of promoter elements which thus can be cloned into a plasmid very easily for further investigations. Promoter analyses of three clones in the chloramphenicol acetyltransferase transient expression assay demonstrated that the recombination with cellular DNA enables the initially defective SV40 promoter to express at wild type levels. PMID- 2545706 TI - Spin-trapping and human neutrophils. Limits of detection of hydroxyl radical. AB - Using the spin trap, 5,5-dimethyl-1-pyrroline-1-oxide (DMPO) and an excess of dimethyl sulfoxide, we previously reported that in the absence of an exogenous iron catalyst, human neutrophils will not generate hydroxyl radical, manifested as the catalse-inhibitable methyl radical spin-trapped adduct, 2,2,5-trimethyl-1 pyrrolidinyloxy (DMPO-CH3) (Britigan, B. E., Rosen, G. M., Chai, Y., and Cohen, M. S. (1986) J. Biol. Chem. 261, 4426-4431). However, superoxide destroys the preformed hydroxyl radical spin-trapped adduct, 2,2-dimethyl-5-hydroxy-1 pyrrolidinyloxy (DMPO-OH), and DMPO-CH3. The present study was undertaken to better resolve the limits of sensitivity of the spin-trapping method. Photolytically generated DMPO-CH3 and DMPO-OH slowly decomposed in the presence of a low flux (1 microM/min) of enzymatically (xanthine/xanthine oxidase) generated superoxide, but more rapid decomposition of these adducts occurred with higher superoxide flux (5 microM/min). Inclusion of cysteine markedly increased the rate of DMPO-OH and DMPO-CH3 decomposition, masking the effect of superoxide alone. The addition of varying concentrations of superoxide dismutase did not lead to increased formation of DMPO-OH or DMPO-CH3, as should have occurred if these adducts were being destroyed by superoxide. As a positive control, we employed an iron-supplemented system with phorbol 12-myristate 13-acetate stimulated neutrophils or xanthine/xanthine oxidase to generate DMPO-CH3. Addition of superoxide dismutase increased the magnitude of DMPO-CH3, primarily by increasing the rate of hydrogen peroxide formation, and to a lesser extent by prolonging the half-life of DMPO-CH3. Although spin-trapped adducts can be destroyed by a high concentration of superoxide, or by lower concentrations of superoxide in the presence of thiol-containing compounds, our results demonstrate that such decomposition does not interfere with the ability of the spin-trapping method to detect hydroxyl radical generated by human neutrophils. These data do not support the capacity of neutrophils to generate hydroxyl radical in the absence of an exogenous Haber-Weiss catalyst. PMID- 2545707 TI - Insulin-regulated glucose uptake in rat adipocytes is mediated by two transporter isoforms present in at least two vesicle populations. AB - We have recently described a monoclonal antibody (1F8) that recognizes a form of glucose transporter unique to fat and muscle (James, D. E., Brown, R., Navarro, J., and Pilch, P. F. (1988) Nature 333, 183-185), tissues that respond acutely to insulin by markedly increasing their glucose uptake. Here, we report that rat adipocytes possess two immunologically distinct glucose-transporters: one recognized by 1F8, and one reactive with antibodies raised against the human erythrocyte glucose transporter. Immunoadsorption experiments indicate that these glucose transporters reside in different vesicle populations and that both transporter isoforms translocate from intracellular sites to the plasma membrane in response to insulin. The insulin-regulatable transporter resides in a unique vesicle that comprises 3% or less of the low density microsomes of fat cells and has a limited protein composition that does not include the bulk of another translocatable protein, the insulin-like growth factor II receptor. Immunoprecipitation with 1F8 of microsomal glucose transporters photoaffinity labeled with [3H]cytochalasin B brings down 90% of the label. Similarly, immunoprecipitation with 1F8 of glucose transporters from insulin-stimulated plasma membranes photolabeled with 3-[125I]iodo-4-azidophenethylamido-7-O succinyldeacetyl-f ors kolin, another transporter-selective reagent, results in 75% of the labeled transporter localized in the immunoprecipitate. Thus, insulin action involves the combined effect of translocation from at least two vesicle pools each containing different glucose transporters. The 1F8-reactive transporter comprises the majority of the total transporter pool that is responsible for the insulin-induced increase in glucose transporter number. PMID- 2545708 TI - Atrial natriuretic factor-induced egression of cyclic guanosine 3':5' monophosphate in cultured vascular smooth muscle and endothelial cells. AB - The induction of cyclic GMP formation in target tissues, i.e. vascular smooth muscle and endothelial cells, by atrial natriuretic factor is followed by its egression into plasma and urine. Since the extracellular appearance of this cyclic nucleotide is used as a marker of atrial natriuretic factor's biological activity, the present study was designed to investigate the characteristics of its egression to the extracellular fluid. In contrast to cyclic AMP, whose profile of egression in time closely follows its intracellular levels, cyclic GMP egression begins with the intracellular decline but continues for a prolonged period, even accumulating for up to 24 h in the extracellular medium. The relative egression of cyclic GMP decreases slightly in the presence of phosphodiesterase inhibitors. On the other hand, the process is sensitive to temperature, inhibited by such agents as probenecid, and occurs against a gradient of 7 orders of magnitude. Large increases of cyclic AMP, as induced by forskolin, can effectively compete for the cyclic GMP transport system, resulting in a 3-fold rise in intracellular cyclic GMP levels, which corresponds to a 3 fold decline of extracellular accumulation. Although the biological significance of cyclic GMP egression is unknown, the results of this study suggest that the process may be one of the significant contributors to the control of cyclic GMP levels in the cell with potential physiological consequences. PMID- 2545709 TI - Inhibitory diffusible factor 45 bifunctional activity. As a cell growth inhibitor and as an insulin-like growth factor I-binding protein. AB - From medium conditioned by 3T3 cells, we had previously purified to apparent homogeneity a novel inhibitory diffusible factor of 45 kDa (IDF45), and then determined the amino-terminal sequence. IDF45 prevented reversibly the growth of chick embryo fibroblast (CEF). In these cells, DNA synthesis stimulated by 1% serum was 50% inhibited in the presence of 45 ng/ml (1 nM) IDF45. In the present article, we show that, in CEF, DNA synthesis stimulated by IGF-I was 100% inhibited in the presence of purified IDF45. Furthermore, the 45-kDa protein (IDF45) was, after Western blotting, able to bind IGF-I. The inhibitory effect of IDF45 upon serum stimulation did not seem to be the result of its inhibitory activity upon IGF-I stimulation, since stimulation by IGF-I and serum were additive. Moreover, it was possible to dissociate the two inhibitory effects: when added to v-src transformed CEF, IDF45 was able to 100% inhibit stimulation induced by IGF-I and was unable to significantly decrease stimulation induced by serum, as was previously observed. Taken together, our results strongly suggest that IDF45 has two distinct functions, one of which was to bind IGF-I and the other to inhibit serum stimulation. Indeed, it was impossible to separate the two functions when IDF45 was purified by cation exchange fast protein liquid chromatography, a method very different from reverse-phase fast protein liquid chromatography previously used for purification to apparent homogeneity of IDF45. On the other hand, if the IGF binding activity and inhibitory activity effect upon serum stimulation were carried by two different proteins, the presence of IGF-I (in conditions where most of the 45-kDa proteins were bound to IGF-I) should not have affected the activity of the molecule inhibiting serum stimulation. However, we observed the contrary: when IDF45 was bound to IGF-I, it lost its inhibitory effect upon stimulation induced by serum. This suggests that the two activities occurred on the same protein and that IDF45 is a bifunctional protein. PMID- 2545710 TI - Proteolysis in heat-stressed HeLa cells. Stabilization of ubiquitin correlates with the loss of proline endopeptidase. AB - When intact HeLa cells were incubated at 45 degrees C, there was progressive inactivation of proline endopeptidase. Rapid loss of the enzyme did not occur in extracts maintained at 45 degrees C. Since Western blots of sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels showed no decrease in the immunoreactive 70-kDa proline endopeptidase band, its in vivo disappearance apparently results from irreversible denaturation or modification. Loss of proline endopeptidase activity was paralleled by reduced degradation of injected ubiquitin and bovine serum albumin. In contrast, proteolysis of injected lysozyme or pancreatic trypsin inhibitor was barely affected. Electrophoretic analysis of ubiquitin or bovine serum albumin retrieved from heated HeLa cells showed that the injected proteins were intact. Thus, the presence of proline endopeptidase appears to be required for initial cleavage of these two substrates, but it has not been shown that the enzyme is directly responsible. Selective stabilization of a subset of the injected proteins does, however, demonstrate the existence of distinct proteolytic pathways in HeLa cytosol. PMID- 2545711 TI - Neutral sphingomyelinase from human urine. Purification and preparation of monospecific antibodies. AB - A neutral sphingomyelinase which cleaves phosphorylcholine from sphingomyelin at a pH optima of 7.4 was purified 440-fold to apparent homogeneity from normal human urine concentrate employing Sephadex G-75 column chromatography, preparative isoelectric focusing, and sphingosylphospholcholine CH-Sepharose column chromatography. The enzyme is composed of a single polypeptide whose apparent molecular weight is 92,000. Analytical isoelectric focusing revealed that the pI of this enzyme is 6.5. Purified neutral sphingomyelinase was devoid of beta-galactosidase and beta-N-acetylglucosaminidase activity originally present in the urine concentrate. The purified neutral sphingomyelinase (N-SMase) had low levels of phospholipase A1 and A2 activity when phosphatidylcholine was used as a substrate and detergents were included in the assay mixture. However, it had no phospholipase activity toward phosphatidylglycerol and sphingomyelin at pH 4.5 irrespective of the presence or absence of detergents. Monospecific polyclonal antibodies raised against N-SMase immunoprecipitated approximately 70% of N-SMase activity from urine, human kidney proximal tubular cells, and partially purified membrane-bound N-SMase from these cells. Western immunoblot assays revealed that the monospecific polyclonal antibody against urinary N-SMase recognized both the urinary N-SMase and the membrane-bound N-SMase. Because this enzyme is distinct biochemically and immunologically as compared to acid sphingomyelinase (EC 3.1.4.12), we would like to assign it an enzyme catalog number of EC 3.1.4.13. The availability of N-SMase and corresponding antibody will be useful in studying various aspects of this enzyme in biological systems. PMID- 2545712 TI - Palmitylation of viral membrane glycoproteins takes place after exit from the endoplasmic reticulum. AB - Palmitylation of vesicular stomatitis virus G and Sindbis virus E1 glycoproteins has been studied in relation to the transport from the endoplasmic reticulum (ER) to the Golgi complex. Incubation of infected cells at 15 degrees C prevents the transport of newly synthesized membrane proteins from the ER to the Golgi (Saraste, J., and Kuismanen, E. (1984) Cell 38, 535-549). In these conditions, also palmitylation of G protein and of E1 glycoprotein is blocked. When the transport is restored by increasing the temperature, palmitylation occurs quickly and is followed by the complete trimming of peripheral mannose residues due to mannosidase I (a putative cis-Golgi function). Immunofluorescence analysis showed that the G glycoprotein accumulated at 15 degrees C in structures distinct from both ER and Golgi. These studies suggest that transport from the ER to the cis Golgi involves intermediate compartments. PMID- 2545713 TI - Insulin and insulin-like growth factor I (IGF I) stimulate phosphorylation of a Mr 175,000 cytoskeleton-associated protein in intact FRTL5 cells. AB - FRTL5 rat thyroid cells possess separate high affinity receptors for insulin and insulin-like growth factor I (IGF I) that undergo beta-subunit phosphorylation upon interaction with the specific ligand. Phosphorylation is rapid and dose dependent and occurs primarily on tyrosine residues. Within 2 min, both insulin and IGF I also give rise to a Mr 175,000 phosphoprotein (pp175) that can be immunoprecipitated by anti-phosphotyrosine antibody (alpha-Tyr(P]. Phosphorylation of pp175 occurs on serine and threonine as well as tyrosine residues. When FRTL5 cells are solubilized with 1% Triton X-100, alpha-Tyr(P) immunoprecipitates phosphorylated insulin and IGF I receptors but little pp175 from the Triton-soluble fraction. After treatment of the Triton-insoluble portion with 1% sodium dodecyl sulfate at 100 degrees C, pp175 can be identified by immunoprecipitation with alpha-Tyr(P). The fraction of FRTL5 cells that remains after extraction of an attached monolayer with 1% Triton for 5 min at 22 degrees C contains most of the cytoskeleton and also nuclei. Extraction of this 32P labeled cytoskeleton preparation with sodium dodecyl sulfate followed by alpha Tyr(P) immunoprecipitation results in almost complete recovery of the pp175 content of the cells. When a nuclear fraction was prepared from FRTL5 cells by differential centrifugation, pp175 was not found in the nuclear pellet from labeled cells, but greater than 80% of pp175 was recovered in the supernatant. We conclude that pp175 is a common substrate for insulin and IGF I receptor kinases which, in FRTL5 cells, is associated with the cytoskeleton. It is suggested that phosphorylation of proteins associated with cytoskeletal elements could be involved in insulin and IGF I action in cells. PMID- 2545714 TI - Phosphorylation sites on two domains of the beta 2-adrenergic receptor are involved in distinct pathways of receptor desensitization. AB - Continuous exposure of cells to neurotransmitter or hormone agonists often results in a rapid desensitization of the cellular response. For example, pretreatment of Chinese hamster fibroblasts (CHW cells) expressing beta 2 adrenergic receptors (beta 2AR) with low (nanomolar) concentrations of isoproterenol, a beta-adrenergic agonist, causes decreases in the sensitivity of the cellular adenylyl cyclase response to the agonist, without changing the maximal responsiveness. In contrast, exposure of CHW cells to high (micromolar) concentrations of isoproterenol results in decreases in both sensitivity and the maximal responsiveness to agonist. To explore the role(s) of receptor phosphorylation in these processes, we expressed in CHW cells three mutant beta 2AR genes encoding receptors lacking putative phosphorylation sites for the cAMP dependent protein kinase A and/or the cAMP-independent beta 2AR kinase. Using these mutants we found that exposure of cells to low concentrations of agonist appears to preferentially induce phosphorylation at protein kinase A sites. This phosphorylation correlates with the decreased sensitivity to agonist stimulation of the adenylyl cyclase response. At higher agonist concentrations phosphorylation on both the beta 2AR kinase and protein kinase A sites occurs, and only then is the maximal cyclase responsiveness elicited by agonist reduced. We conclude that low or high concentrations of agonist elicit phosphorylation of beta 2AR on distinct domains, with different implications for the functional coupling of the receptors with effector molecules. PMID- 2545715 TI - Identification of integrin-like matrix receptors with affinity for interstitial collagens. AB - Antibodies to a rat liver membrane glycoprotein with an Mr of 115,000 (nonreduced) inhibited the attachment of rat hepatocytes and primary rat heart fibroblasts to both collagen and fibronectin. The Mr 115,000 glycoprotein cross reacted immunologically with the beta 1-chain of the rat hepatocyte fibronectin receptor (HFNR), and the two proteins showed identical peptide maps after proteolytic cleavage. It was concluded that the Mr 115,000 protein was similar or identical to the beta 1-chain of Arg-Gly-Asp (RGD)-directed matrix receptors. Although collagen type I contains several RGD sequences, the attachment of hepatocytes and fibroblasts to collagen type I was not inhibited by the synthetic peptide GRGDTP in concentrations that blocked adhesion to fibronectin. Furthermore, hepatocytes adhered equally well to collagen fragments, generated by cyanogen bromide cleavage, lacking RGD sequences as to fragments containing this sequence. Antibodies to the Mr 115,000 protein inhibited the adhesion of hepatocytes to both types of collagen fragments. Taken together, these data indicate the presence of collagen receptors that share the beta-subunit with the HFNR but that are not directed to RGD sequences. Tentative alpha-chains of the collagen matrix receptor complex were isolated by immunoprecipitation of surface 125I-labeled fibroblast membrane proteins purified by affinity chromatography on immobilized collagen type I. Data are presented indicating that proteins with Mr around 145,000 and 170,000 (nonreduced) are associated in noncovalently linked complexes with the Mr 115,000 protein. These complexes have affinity for collagen and thus have properties expected for integrin-like collagen receptors. PMID- 2545716 TI - 8-phenyltheophylline as an inhibitor of cyclic AMP hydrolysis by cyclic nucleotide phosphodiesterase. AB - 1. The inhibition, by theophylline and 8-phenyltheophylline, of cAMP hydrolysis by cyclic nucleotide phosphodiesterase from rat fat cells, abdominal aorta, gastrocnemius muscle, erythrocytes and cerebrum was examined. 2. Theophylline was an approximately equieffective inhibitor of cAMP hydrolysis in all tissue extracts. In contrast, 8-phenyltheophylline was a markedly more effective inhibitor of cAMP breakdown in erythrocytes and skeletal muscle than in smooth muscle, brain and fat cells. The 8-phenyl substituted compound was a more potent inhibitor in erythrocytes and skeletal muscle than theophylline. 3. 8 phenyltheophylline has been postulated to be a very selective adenosine receptor antagonist, the present study indicates, that in some tissues 8 phenyltheophylline is not so selective as an adenosine receptor antagonist as has previously been suggested. 4. Analysis of cyclic AMP breakdown by cyclic nucleotide phosphodiesterase in fat cells and erythrocytes demonstrated the presence of high and low affinity forms. 5. Theophylline was slightly more effective as an inhibitor of the high than of the low affinity forms in both tissues. 8-phenyltheophylline was weakly effective as an inhibitor of all isoenzymes from fat cells and selectively inhibited the high affinity phosphodiesterase from erythrocytes. 6. The results suggest that 8 phenyltheophylline is a selective inhibitor of a cyclic nucleotide phosphodiesterase with a high affinity for cAMP, which has relatively greater activity in erythrocytes, and presumably in skeletal muscle, than in other tissues such as fat cells. PMID- 2545717 TI - The effects of purine nucleotides on transmission in vesical parasympathetic ganglia of the cat. AB - 1. In adult cats, postganglionic nerve fibres on the surface of the bladder were isolated and multiunit activity of these fibres was recorded. In these cats, the urinary bladder was cannulated and intravesical pressure was also recorded. 2. ATP, APPCP, ADP, AMP and adenosine depress transmission in vesical parasympathetic ganglia equipotently; however, 2-chloroadenosine was 10-fold more potent than ATP. 3. 2-chloroadenosine, ATP, ADP, AMP, adenosine and APPCP inhibit neurally evoked bladder contractions in the same order or potency with which they depress pelvic ganglionic transmission; however, adenine, inosine, IMP and ITP were ineffective. 4. 3',5'-cyclic AMP and dibutyryl cAMP produced little or no effect on bladder activity. 5. ATP and APPCP produced a transient rise in intravesical pressure at doses 2 to 50 times the dose needed for inhibition, presumably through ATP (P2) receptors. APPCP was 10 to 20 times more potent in exciting the bladder than ATP. 6. Theophylline and caffeine effectively antagonized purinergic effects mediated through adenosine (P1) receptors on both pelvic ganglia and bladder smooth muscle. 7. ATP inhibition of TMA-evoked bladder contractions and postganglionic nerve firing suggests that purinergic inhibition occurs, at least in part, at a postsynaptic site in the ganglia. PMID- 2545718 TI - Partial agonistic activity of GR43175 at the inhibitory prejunctional 5-HT1-like receptor in rat kidney. AB - 1. The pharmacological effect of the selective 5-HT receptor agonist GR43175 has been studied in the isolated perfused rat kidney with particular reference to the inhibitory prejunctional 5-HT1-like receptor. 2. GR43175 (1 x 10(-8) to 1 x 10( 5) M) inhibited the electrically evoked release of tritium from renal noradrenergic nerves but did not affect the basal (non-stimulated) release of tritium or perfusion pressure. 3. The inhibitory prejunctional action of GR43175, as well as that of 5-HT, was blocked by methiothepin, suggesting that both agonists act at the inhibitory 5-HT1-like site. 4. GR43175, relative to 5-HT, acted as a weak partial agonist at the inhibitory prejunctional 5-HT1-like receptor and antagonized 5-HT-induced inhibition of tritium release, suggesting again that a common site of action is involved. 5. The results support the conclusion that the 5-HT1-like receptor activated by GR43175 in the rat kidney is very similar to that identified in dog saphenous vein and certain other tissues. PMID- 2545719 TI - Characterization of tissue from the bone-polymethylmethacrylate interface in a rat experimental model. Demonstration of collagen-degrading activity and bone resorbing potential. AB - In previous studies, we described a layer of tissue that formed around methylmethacrylate cement that had been implanted into the posterior cervical spine of dogs. We are now reporting on a rat model in which we induced, in the interface between the bone of the posterior elements of the dorsal spine and methylmethacrylate, the formation of a layer of tissue that was morphologically similar to the tissue that had been produced in the dogs. As in the dogs, we noted macrophages and giant cells and we demonstrated that the interface tissue synthesized several basement-membrane components (type-IV collagen, laminin, and fibronectin). In addition, we demonstrated the synthesis of an additional extracellular-matrix protein--type-VI collagen. We also showed that extracts of organ cultures of tissue from the rat model degraded type-I collagen into three quarter and one-quarter-length fragments. Such enzymatic activity is characterized of mammalian collagenase, an enzyme that is known to play a critical role in the resorption of bone. PMID- 2545720 TI - A histological study of calcium pyrophosphate dihydrate crystal-deposition disease. AB - Synovial, meniscal, articular cartilage, and other connective tissue from fifty seven patients who had calcium pyrophosphate dihydrate crystal-deposition disease was examined by light microscopy, electron microscopy, and electron-probe microanalysis. Safranin O-positive hypertrophic chondrocytes that contained proteoglycans were observed in the tissues of each patient. Microcrystals that were suggestive of early precipitation of crystals were found in the degenerating matrix surrounding hypertrophic chondrocytes. The matrix contained electron-dense amorphous material, including proteoglycans and debris of cellular components. The microcrystals were often seen in contact with degenerating collagen fibers. There was never any histological evidence of formation of crystal in the areas that had no hypertrophic chondrocytes. Chondrocytes of this kind, surrounded by characteristic degenerating matrix, were never observed in the articular tissue from sixty-one patients who had only osteoarthritis. On the basis of our results, we speculate that electron-dense amorphous material containing proteoglycans and debris of cellular components, and the degenerating collagen fibers that were seen around the hypertrophic chondrocytes, may play important roles in the formation of calcium pyrophosphate dihydrate crystals. PMID- 2545721 TI - Autoantibody formation in burn patients after inhibition of suppressor T cell activity with polymyxin B. AB - After thermal injury, treatment with polymyxin B blocks suppressor T cell activity by uncoupling endotoxin-mediated T cell activation, but the effect on autoantibody formation is unknown. We examined the presence of antinuclear antibodies to native DNA; to soluble antigens Ro/SSA, La/SSB, Sm, nRNP; and to antiepithelial antibodies in 12 burn patients before and after treatment with polymyxin B and in 24 samples from control burn patients. Low titer antinuclear antibody activity was detected in 25% of pretreatment and 78% of posttreatment samples (p less than 0.01) and in 16.7% of control patients. One polymyxin B treated patient had a significant antinuclear antibody titer both before and after treatment. Antiepithelial antibodies were detected in 16.7% of early polymyxin B-treated samples and 11.1% of late samples (p less than 0.05) but were also present in 20.8% of controls. Antibodies to native DNA, Ro/SSA, La/SSB, Sm, and nRNP were not detected in any sera. PMID- 2545722 TI - Multiple tumor emboli after lung resection. AB - A patient with carcinoma of the lung underwent a left lower lobectomy. For technical difficulties the pulmonary vein was not ligated prior to extensive manipulations of the involved lobe. Following the pulmonary surgery the patient sustained a massive aortic occlusion by a tumor embolus, that was removed by bilateral femoral embolectomies. Three additional documented episodes of peripheral arterial emboli subsequently took place, two of which were tumoral. One tumor embolus into the carotid artery territory eventually caused metastatic spread in the brain. All peripheral emboli were successfully treated by embolectomy. This unique display of multiple tumor emboli, following lung resection for carcinoma, reemphasises the significance of early interruption of the pulmonary vein, in an attempt to reduce the incidence of tumor emboli. PMID- 2545723 TI - Recent developments in the cell biology of basic fibroblast growth factor. PMID- 2545724 TI - In vitro import of cytochrome c peroxidase into the intermembrane space: release of the processed form by intact mitochondria. AB - Cytochrome c peroxidase (CCP) is a nuclearly encoded hemoprotein located in the intermembrane space (IMS) of Saccharomyces cerevisiae mitochondria. Wild-type preCCP synthesized in rabbit reticulocyte lysates, however, was inefficiently translocated into isolated mitochondria and was inherently resistant to externally added proteases. To test whether premature heme addition to the apoprecursor was responsible for the protease resistance and the inability to import preCCP, site-directed mutagenesis was used to replace the axial heme ligand (His175) involved in forming a pseudo-covalent link between the heme iron and CCP. Mutant proteins containing Leu, Arg, Met, or Pro at residue 175 of mature CCP were sensitive to proteolysis and were imported into isolated mitochondria as judged by proteolytic processing of the precursor. The inhibition of wild-type CCP translocation across the outer membrane may result from the inability of the heme-containing protein to unfold during the translocation process. Although the protease responsible for cleaving preCCP to its mature form is believed to be located in the IMS, most of the processed CCP was located in the supernatant rather than the mitochondrial pellet. Since the outer membranes were shown to be intact, the anomalous localization indicated that preCCP may not have been completely translocated into the IMS before proteolytic processing or that conformationally labile proteins may not be retained by the outer membrane. Proteolytic maturation of preCCP also occurred in the presence of valinomycin, suggesting that the precursor may be completely or partially translocated across the outer mitochondrial membrane independent of a potential across the inner mitochondrial membrane. PMID- 2545725 TI - Subcellular distribution of the 1,4-dihydropyridine receptor in rabbit skeletal muscle in situ: an immunofluorescence and immunocolloidal gold-labeling study. AB - The subcellular distribution of the 1,4-dihydropyridine receptor was determined in rabbit skeletal muscle in situ by immunofluorescence and immunoelectron microscopy. Longitudinal and transverse cryosections (5-8 microns) of rabbit gracilis muscle were labeled with monoclonal antibodies specific against either the alpha 1-subunit (170,000-D polypeptide) or the beta-subunit (52,000-D polypeptide) of the 1,4-dihydropyridine receptor by immunofluorescence labeling. In longitudinal sections, specific labeling was present only near the interface between the A- and I-band regions of the sarcomeres. In transverse sections, specific labeling showed a hexagonal staining pattern within each myofiber however, the relative staining intensity of the type II (fast) fibers was judged to be three- to fourfold higher than that of the type I (slow) fibers. Specific immunofluorescence labeling of the sarcolemma was not observed in either longitudinal or transverse sections. These results are consistent with the idea that the alpha 1-subunit and the beta-subunit of the purified 1,4-dihydropyridine receptor are densely distributed in the transverse tubular membrane. Immunoelectron microscopical localization with a monoclonal antibody to the alpha 1-subunit of the 1,4-dihydropyridine receptor showed that the 1,4-dihydropyridine receptor is densely distributed in the transverse tubular membrane. Approximately half of these were distributed in close proximity to the junctional region between the transverse tubules and the terminal cisternae. Specific labeling was also present in discrete foci in the subsarcolemmal region of the myofibers. The size and the nonrandom distribution of these foci in the subsarcolemmal region support the possibility that they correspond to invaginations from the sarcolemma called caveolae. In conclusion, our results demonstrate that the 1,4 dihydropyridine receptor in skeletal muscle is localized to the transverse tubular membrane and discrete foci in the subsarcolemmal region, possibly caveolae but absent from the lateral portion of the sarcolemma. PMID- 2545726 TI - Characterization of 125I-tissue plasminogen activator binding to cerebellar granule neurons. AB - Mouse cerebellar cells in culture secrete tissue plasminogen activator (tPA) into the culture medium. Fibrin overlays have shown tPA to be associated with granule neurons in these cultures. This cell associated tPA can be displaced by extensive washing of the cells or by a brief lowering of the pH (less than 4), which leads to a loss of fibrinolytic activity by the cells. Incubation of these fibrinolytically inactive cells with exogenously added murine tPA leads to the restoration of the fibrinolytic activity, indicating the presence of tPA binding sites on these granule neurons. Using 125I-tPA, the binding to cerebellar granule neurons is rapid, saturable, specific, high affinity (Kd = 50 pM) and reversible. Both murine and human tPA compete with 125I-tPA for binding, with both murine and human urokinase (uPA) as well as human thrombin and plasminogen fail to compete. Neither the catalytic site nor the carbohydrate moiety of tPA appear to be involved in the binding, since both diisopropyl-fluorophosphate-treated tPA and endoglycosidase-H-treated tPA compete with 12I-tPA for binding. Furthermore, epidermal growth factor does not compete well with tPA for binding even at a 10:1 molar excess, suggesting that the epidermal growth factor-like (EGF) domain of tPA may not be involved in the binding mechanism. Autoradiography and antibody immunofluorescence show the specific tPA binding is to granule neurons in these cultures. Thus, granule neurons possess tPA receptors on their surface, where this protease binds retaining is functional activity and may play a role in cell and axon migration. PMID- 2545727 TI - Mechano-chemical control of human endothelium orientation and size. AB - Human umbilical vein endothelial cells (EC) were grown on elastic silicone membranes subjected to cyclic stretch, simulating arterial wall motion. Stretching conditions (20% amplitude, 52 cycle/min) stimulated stress fiber formation and their orientation transversely to the strain direction. Cell bodies aligned along the same axis after the actin cytoskeleton. EC orientation response was inhibited by the adenylate cyclase activator, forskolin (10(-5) M), which caused stress fiber disassembly and the redistribution of F-actin to the cortical cytoplasm. Preoriented EC depleted of stress fibers by forskolin treatment retained their aligned state. Thus, stress fibers are essential for the process of EC orientation induced by repeated strain, but not for the maintenance of EC orientation. The monolayer formed by EC grown to confluence in conditions of intermittent strain consisted of uniform elongated cells and was resistant to deformation. In contrast, the monolayer assembled in stationary conditions was less compliant and exposed local denudations on initiation of stretching. When stretched in the presence of 10(-5) M forskolin it rapidly (3-4 h) reestablished integrity but gained a heterogeneous appearance since denuded areas were covered by giant cells. The protective effect of forskolin was because of the stimulation of EC spreading. This feature of forskolin was demonstrated while studying its action on EC spreading and repair of a scratched EC monolayer in conventional culture. Thus mechanical deformation and adenylate cyclase activity may be important factors in the control of endothelium morphology in human arteries. PMID- 2545728 TI - Von Willebrand factor promotes endothelial cell adhesion via an Arg-Gly-Asp dependent mechanism. AB - Von Willebrand factor (vWF) is a constitutive and specific component of endothelial cell (EC) matrix. In this paper we show that, in vitro, vWF can induce EC adhesion and promote organization of microfilaments and adhesion plaques. In contrast, human vascular smooth muscle cells and MG63 osteosarcoma cells did not adhere and spread on vWF. Using antibodies to the beta chains of fibronectin (beta 1) and vitronectin (beta 3) receptors it was found that ECs adherent to vWF show clustering of both receptors. The beta 1 receptor antibodies are arranged along stress fibers at sites of extracellular matrix contact while the beta 3 receptor antibodies were sharply confined at adhesion plaques. ECs release and organize endogenous fibronectin early during adhesion to vWF. Upon blocking protein synthesis and secretion, ECs can equally adhere and spread on vWF but, while the beta 3 receptors are regularly organized, the beta 1 receptors remain diffuse. This suggests that the organization of the beta 1 receptors depend on the release of fibronectin and/or other matrix proteins operated by the same cell. Antibodies to the beta 3 receptors fully block EC adhesion to vWF and detach ECs seeded on this substratum. In contrast, antibodies to the beta 1 receptors are poorly active. Overall these results fit with an accessory role of beta 1 receptors and indicate a leading role for the beta 3 receptors in EC interaction with vWF. To identify the EC binding domain on vWF we used monoclonal antibodies produced against a peptide representing the residues Glu1737-Ser1750 of the mature vWF and thought to be important in mediating its binding to the platelet receptor glycoprotein IIb-IIIa. We found that the antibody that recognizes the residues 1,744-1,746, containing the Arg-Gly-Asp sequence, completely inhibit EC adhesion to vWF whereas a second antibody recognizing the adjacent residues 1,740-1,742 (Arg-Gly-Asp-free) is inactive. Both antibodies do not interfere with EC adhesion to vitronectin. This defines the molecular domain on vWF that is specifically recognized by ECs and reaffirms the direct role of the Arg-Gly-Asp sequence as the integrin receptor recognition site also in the vWF molecule. PMID- 2545729 TI - The primary structure of the VLA-2/collagen receptor alpha 2 subunit (platelet GPIa): homology to other integrins and the presence of a possible collagen binding domain. AB - VLA-2 (also called gpIa/IIa on platelets) is a collagen receptor with a unique alpha subunit and a beta subunit common to other adhesion receptors in the VLA/integrin family. Multiple cDNA clones for the human VLA-2 alpha 2 subunit have been selected from a lambda gtll library by specific antibody screening. The 5,374-bp nucleotide sequence encoded for 1,181 amino acids, including a signal peptide of 29 amino acids followed by a long extracellular domain (1,103 amino acids), a transmembrane domain, and a short cytoplasmic segment (22 amino acids). Direct sequencing of purified alpha 2 protein confirmed the identity of the 15 NH2-terminal amino acids. Overall, the alpha 2 amino acid sequence was 18-25% similar to the sequences known for other integrin alpha subunits. In particular, the alpha 2 sequence matched other integrin alpha chains in (a) the positions of 17 of its 20 cysteine residues; (b) the presence of three metal-binding domains of the general structure DXDXDGXXD; and (c) the transmembrane domain sequence. In addition, the alpha 2 sequence has a 191-amino acid insert (called the I-domain), previously found only in leukocyte integrins of the beta 2 integrin family. The alpha 2 I-domain was 23-41% similar to domains in cartilage matrix protein and von Willebrand factor, which are perhaps associated with collagen binding. The NH2-terminal sequence reported here for alpha 2 does not match the previously reported alpha 2 NH2-terminal sequence (Takada, Y., J. L. Strominger, and M. E. Hemler. 1987. Proc. Natl. Acad. Sci. USA. 84:3239-3243). Resolution of this discrepancy suggests that there may be another VLA heterodimer that resembles VLA 2 in size but has a different amino acid sequence. PMID- 2545731 TI - Regulation of hepatocyte growth: alpha-1 adrenergic receptor and ras p21 changes in liver regeneration. AB - Catecholamines, acting via the alpha-1 adrenergic receptor, have been demonstrated to influence adult rat hepatocyte DNA synthesis in primary culture and in vivo during liver regeneration following partial hepatectomy (PHX). Earlier investigations have suggested that the alpha-1 effect on DNA synthesis is significant only during the first day following PHX. We examined receptor binding at several early and late time points after surgery, and we observed a significant loss of specific [3H]-prazosin binding to cells isolated from rat livers 48 and 72 hr after PHX. In contrast, the ability of norepinephrine to stimulate inositol phosphate production in isolated cells prelabeled with [3H] myo-inositol was transiently reduced between 8 and 16 hr, when alpha-1 binding capacity was virtually unchanged. This uncoupling of phosphoinositide turnover from binding was preceded by a drop in hepatic membrane ras p21 content, as assayed by liquid competition radioimmunoassay. The loss of immunoreactive p21 from membranes was significant by 2 hr after PHX. These findings suggest a role for alpha-1 receptors and ras protein in the early events of liver regeneration. PMID- 2545730 TI - Spatial segregation of the regulated and constitutive secretory pathways. AB - Recent experiments using DNA transfection have shown that secretory proteins in AtT-20 cells are sorted into two biochemically distinct secretory pathways. These two pathways differ in the temporal regulation of exocytosis. Proteins secreted by the regulated pathway are stored in dense-core granules until release is stimulated by secretagogues. In contrast, proteins secreted by the constitutive pathway are exported continuously, without storage. It is not known whether there are mechanisms to segregate regulated and constitutive secretory vesicles spatially. In this study, we examined the site of insertion of constitutive vesicles and compared it with that of regulated secretory granules. Regulated granules accumulate at tips of processes in these cells. To determine whether constitutively externalized membrane proteins are inserted into plasma membrane at the cell body or at process tips, AtT-20 cells were infected with ts-O45, a temperature-sensitive mutant of vesicular stomatitis virus in which transport of the surface glycoprotein G is conditionally blocked in the ER. After switching to the permissive temperature, insertion of G protein was detected at the cell body, not at process tips. Targeting of constitutive and regulated secretory vesicles to distinct areas of the plasma membrane appears to be mediated by microtubules. We found that while disruption of microtubules by colchicine had no effect on constitutive secretion, it completely blocked the accumulation of regulated granules at special release sites. Colchicine also affected the proper packaging of regulated secretory proteins. We conclude that regulated and constitutive secretory vesicles are targeted to different areas of the plasma membrane, most probably by differential interactions with microtubules. These results imply that regulated secretory granules may have unique membrane receptors for selective attachment to microtubules. PMID- 2545732 TI - Growth-inhibitory effect of TGF-B on human fetal adrenal cells in primary monolayer culture. AB - We examined the effects of transforming-growth factor-B (TGF-B) on growth ([3H] thymidine uptake) and function (dehydroepiandrosterone sulfate [DHAS] and cortisol production) of human fetal zone adrenal cells. Results indicate that TGF B significantly inhibits, in a dose-related manner, both basal and epidermal growth factor (EGF)-stimulated cell growth: IC50 = 0.1-0.25 ng/ml. EGF is ineffective in overcoming the inhibitory effect of TGF-B, suggesting a noncompetitive antagonism between the two factors. Also, the inhibitory effect of TGF-B is additive to that of adrenocorticotropic hormone (ACTH). On the other hand, TGF-B (1 ng/ml) does not significantly change basal or ACTH-stimulated DHAS or cortisol secretion. We conclude that, unlike its effect on other steroid producing cells, TGF-B inhibits growth of fetal zone cells and does not appear to have a significant inhibitory effect on steroidogenesis. PMID- 2545734 TI - Biphasic regulation of macrophage attachment by activators of cyclic adenosine monophosphate-dependent kinase and protein kinase C. AB - A method is described that enabled us to study the adhesiveness of J-774 murine macrophages. Cell attachment was stimulated by activators of kinase C (i.e., phorbol esters) as well as kinase A (cyclic adenosine monophosphate; cAMP). This novel effect of cAMP was observed when its levels were increased via receptor triggering (prostaglandin E1, beta-adrenergic agonists), activation of Ns (cholera toxin), or inhibition of phosphodiesterase (Ro 20-1724) or when the kinase was directly activated by Br8-cAMP. The simultaneous treatment with kinase A and kinase C activators at the time of attachment resulted in a partially additive response. On the other hand, preincubation of the cells in suspension with one of the activators rendered them refractory to subsequent stimulation at the onset of the adhesion assay, whatever agent was used. Such a refractoriness was also observed in cells preincubated with oleoyl-acetyl-glycerol (OAG). On the other hand, when added at the time of attachment, this near-physiological activator of kinase C evoked a biphasic response: the early stimulation of cell attachment was followed by an accelerated rate of "detachment." In conclusion, kinase C and kinase A play a role in the sequence of events leading to cell adhesion. The cross desensitization observed is distal and takes place at or beyond the kinase step. PMID- 2545733 TI - Coupling of bradykinin receptors to phospholipase C in cultured fibroblasts is mediated by a G-protein. AB - In cultured foreskin fibroblasts, bradykinin stimulates inositol phosphate generation, arachidonic acid release, and Na+/H+ exchange, with doses of 1-3 nM yielding half-maximal stimulation. Binding of 3H-bradykinin to these cells demonstrates a single receptor site with a Kd of 2.0 nM and a Bmax of 91 fmoles/mg protein. Bradykinin analogs of the B2 type inhibit this binding. GTP synergizes with bradykinin to stimulate phosphatidylinositol turnover in permeabilized fibroblasts and GTP-gamma-S decreases the Bmax of bradykinin binding to fibroblast membranes, indicating that a G-protein couples the receptor to phospholipase C. Pretreatment of fibroblasts with either cholera or pertussis toxin enhances bradykinin stimulation of inositol phosphate accumulation. PMID- 2545735 TI - Viruses and cytokines: evidence for multiple roles in pancreatic beta cell destruction in type 1 insulin-dependent diabetes mellitus. AB - Insulin-dependent (type 1) diabetes mellitus (IDDM) is due to the selective autoimmune-mediated destruction of pancreatic beta cells possibly initiated by viruses. To elucidate the possible role of viruses and cytokines in the pathogenesis of IDDM, we have examined the effect of reovirus infection on beta cell major histocompatibility complex (MHC) expression and the effect of interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) on beta cell function in vitro. Infection of RIN-m5F (rat insulinoma) cells with reovirus 1 or reovirus-3 was associated with a tenfold increase in class 1 MHC protein and mRNA expression. Reovirus infection did not induced the expression of class 11 MHC by RIN-m5F cells. Exposure of reovirus to ultraviolet light almost completely abolished its ability to induce class 1 MHC protein expression on infected cells. Murine islets cultured for 3 days with IFN-gamma and/or TNF-alpha had a significantly reduced insulin response to glucose, which was more marked with a combination of the cytokines. During 6 days of culture in IFN-gamma plus TNF alpha islets underwent noticeable degeneration associated with an 80% reduction in insulin content. These findings together with previous data suggest viruses and cytokines may have multiple roles in beta cell destruction, indirectly through enhanced MHC protein expression and directly through functional impairment and loss of viability. PMID- 2545736 TI - [Oxygen free radicals and inflammatory diseases of intestines]. AB - Free oxygen radicals (F.O.R.) belong to a very aggressive chemical species derived from molecular oxygen. Their role in inflammation is well established and Polymorphonuclear neutrophils (PMNS) make use of them as antibacterial weapons. Their role has been experimentally demonstrated in numerous ischemia-reperfusion models. Free radical scavengers such as the superoxide dismutase, allopurinol or desferrioxamine can prevent the occurrence of lesions. The essential role of PMNS in these models is demonstrated by the fact that previous depletion of the animal in PMNS also prevents such lesions. Histologically, in these ischemia-reperfusion models, PMNS infiltration may be quantified by assay of myeloperoxidase. In experimental models of inflammatory colitis (acetic acid, bacterial polysaccharides) intestinal wall infiltration by PMNS is a fundamental phenomenon and is also a characteristic of Crohn's disease and exacerbations of Ulcerative Colitis. Thus, it is probable in both disorders that F.O.R. play an important role since steroids inhibit their secretion by PMNS and 5-aminosalicylic acid has been shown to be a F.O.R. scavenger. PMID- 2545737 TI - Anion-cation separations on a mixed bed alumina-silica column. AB - Mixed bed ion-exchange (MBIE) columns containing alumina and silica were evaluated for the simultaneous separation of anion and cation analytes. At the mobile phase pH used alumina provides anion exchange sites while silica provides cation exchange sites. Since alumina and silica exhibit weak acid and base properties, their anion and/or cation exchange properties are pH dependent. Ion exchange capacities, rates of exchange and analyte ion exchange selectivities are also pH dependent. The major mobile phase parameters affecting analyte anion and cation resolution and elution order are pH and type and concentration of counter anion and counter cation, respectively. The weight ratio of the two exchangers and/or the exchange capacities of the two in the column can also be used to alter resolution and elution order. Several examples of the simultaneous separation of inorganic mono- and divalent anions and cations using a single sample injection, a single column and a single detector (conductivity) illustrate the parameters and scope of the alumina-silica MBIE column. PMID- 2545738 TI - New genome types of adenovirus types 1, 3, and 5 isolated from stools of children in Brazil. AB - During an epidemiological survey made in Sao Paulo (Brazil), fecal specimens were periodically collected from 100 randomly chosen babies from birth to the age of 18 months. The stools, routinely collected each month and also collected each time a child presented any sign of disease, were screened for the presence of adenoviruses. Sixteen adenovirus strains, isolated from the stools of healthy and ill children, were characterized by restriction enzyme analysis. Five isolates were from subgenus A, five were from subgenus B, four were from subgenus C, and two were from subgenus D. All but two showed some restriction patterns different from those of the 42 human adenovirus prototypes and all the genome types described up to now. No fastidious adenovirus (subgenus F, serotypes 40 and 41) was encountered in the stools examined. We report here the restriction enzyme analysis of isolates of subgenera B and C. The following new designation genome types are proposed: Ad3e1 (subgenus B) and Ad1d, Ad5a1, and Ad5a2 (subgenus C). PMID- 2545739 TI - Rapid detection of herpes simplex virus with fluorescein-labeled Helix pomatia lectin. AB - The use of fluorescein-conjugated Helix pomatia lectin was shown to be as effective as fluorescein-conjugated monoclonal antibody reagents for the detection and differentiation of herpes simplex virus type 1 and type 2 (HSV-1 and HSV-2) in MRC-5 cell culture. Cells infected with HSV-1 generally displayed a pattern of nongranular or diffuse fluorescence, while cells infected with HSV-2 were identified by the production of fluorescent grains and flecks. This unique nonimmunological reagent, when used in combination with low-speed centrifugation, provides a remarkably specific, sensitive, rapid, and cost-effective means to detect HSV-infected MRC-5 or BHK-21 cells as early as 20 h postinoculation. In contrast to the immunofluorescence method, the serotypes of HSV can be differentiated with only one fluorescein-H. pomatia reagent in MRC-5 cell cultures. PMID- 2545740 TI - Comparison of four charcoal media for the isolation of Bordetella pertussis. AB - Charcoal-horse blood agar with 40 micrograms of cephalexin per ml, charcoal-horse blood agar with 3 micrograms of lincomycin per ml, charcoal agar with 3 micrograms of lincomycin per ml, and Legionella (buffered charcoal-yeast extract) agar with 3 micrograms of lincomycin per ml were compared for isolation of Bordetella pertussis. Charcoal-horse blood agar with 40 micrograms of cephalexin per ml gave the best results, with a B. pertussis recovery rate of 100%. Growth was most rapid and the mean number of colonies was highest on this agar, and growth of pharyngeal flora was completely suppressed. PMID- 2545741 TI - Comparison of methods for determining DNase and phosphatase activities of staphylococci. AB - A greater percentage of DNase-positive strains was detected with DNase test agar than with DNase test agar containing 0.005% methyl green or 0.005% toluidine blue (P less than 0.01). No significant differences were obtained in the percentage of phosphatase-positive strains with the four methods compared. On the basis of ease of use, P agar containing para-nitrophenylphosphate disodium (0.495 mg/ml) would be the preferred method for determining phosphatase activity of staphylococci. PMID- 2545742 TI - In situ hybridization for quantitative assay of infectious hepatitis A virus. AB - A method of in situ hybridization using single-stranded RNA probes of opposite polarity for quantitative enumeration of hepatitis A virus (HAV) in infected cells has been developed. Kinetic experiments showed that foci of infected cells appeared as early as day 2 postinfection. The absence of foci in cells examined immediately after virus adsorption indicated that foci detected subsequently were related to viral replication. Foci were detected by hybridization with RNA probes complementary to HAV genomic RNA but not with RNA probes identical to HAV genomic RNA. The number of foci observed was linearly related to the HAV dose inoculated. Focus formation was reduced when a virus inoculum was pretreated with guinea pig anti-HAV hyperimmune serum but not when it was pretreated with preimmune serum. The high resolution of hybridization signals and relative rapidity of the test indicated that this technique will be useful for measuring serum neutralizing antibodies and for quantitative assay of infectious HAV. PMID- 2545743 TI - Noninfectious rotavirus (strain RRV) induces an immune response in mice which protects against rotavirus challenge. AB - We found that female adult mice parenterally inoculated with noninfectious rotavirus (simian strain RRV) developed virus-specific neutralizing antibodies in the serum; newborn mice from these dams were protected against RRV-induced gastroenteritis. In addition, mice parenterally inoculated with noninfectious RRV developed virus-specific cytotoxic T-lymphocyte precursors in the spleen. Replication of rotavirus in intestinal epithelial cells was apparently not required to induce rotavirus-neutralizing antibodies or rotavirus-specific cytotoxic T lymphocytes. Parenteral immunization of infants and young children with noninfectious rotaviruses may induce an immune response which protects against rotavirus challenge. PMID- 2545745 TI - The National Health Objectives Act. A proposal by the Association of State and Territorial Health Officials. PMID- 2545744 TI - Rapid diagnosis of acute Epstein-Barr virus infection by an indirect enzyme linked immunosorbent assay for specific immunoglobulin M (IgM) antibody without rheumatoid factor and specific IgG interference. AB - An indirect enzyme-linked immunosorbent assay (ELISA) for detection of Epstein Barr virus-specific immunoglobulin M (IgM) antibody was developed with commercial reagents. Sera containing rheumatoid factor (RF) (as little as 0.5 IU/ml) coupled with specific IgG resulted in false-positives in the ELISA. This interference was eliminated by the use of anti-human IgG antibodies to remove RF and IgG. Thus, pathogen-specific IgG complexes to which IgM-RF could be bound during the subsequent test were inhibited, and competition between specific IgG and IgM was also prevented. Of the 1,672 serum specimens tested, 353 were found to be Epstein Barr virus IgM antibody positive by indirect immunofluorescence (IF). Compared with the IF test, the ELISA showed 96.6% sensitivity, 99.7% specificity, and 99% accuracy. Further evidence indicated that most of the 12 ELISA false-negatives were IF false-positives. There was a linear correlation between mean ELISA values and increasing IF titers (r = 0.96). However, the IF test has the disadvantages that it lacks automated reading and requires considerable technical expertise, both of which restrict the range of laboratories performing the test. The indirect ELISA has the advantages that it is simple and rapid and can be automated. All the reagents used in this assay are commercially available, have been prestandardized, and are stable. PMID- 2545746 TI - GABA-immunoreactive neurons and terminals in the lateral cervical nucleus of the cynomolgus monkey. AB - An antiserum against the inhibitory transmitter substance gamma-aminobutyric acid (GABA) was used to investigate the distribution of GABAergic nerve terminals and cell bodies in the lateral cervical nucleus (LCN) of the cynomolgus monkey. Light microscopic immunohistochemistry demonstrated GABA-immunoreactive puncta, suggestive of nerve terminals, scattered throughout the LCN. The terminal-like profiles are often present along the somata of unlabeled neurons, but most are located in the neuropil. GABA-immunoreactive neurons are present in the LCN, but constitute a very small number of the LCN neurons. Electron microscopy showed that the GABA-positive neurons are small with a relatively large nucleus. They are contacted by few somatic boutons. Numerous GABA-immunoreactive terminals containing densely packed round to oval synaptic vesicles were also found. Most GABA-positive terminals make synaptic contact with dendrites, but synapses with cell bodies are also present. Synaptic contacts between labeled and unlabeled terminals were not observed. Some GABA-positive terminals make contact with GABA positive neurons. The present findings suggest that GABA is a major inhibitory transmitter substance in the LCN of the monkey. However, in comparison with other somatosensory relay nuclei, there are few GABA-immunoreactive neurons in the LCN. This may imply that the GABA-positive neurons branch extensively in the LCN or that an extrinsic source of GABAergic input exists. PMID- 2545747 TI - Human reticular formation: cholinergic neurons of the pedunculopontine and laterodorsal tegmental nuclei and some cytochemical comparisons to forebrain cholinergic neurons. AB - Choline acetyltransferase immunohistochemistry showed that the human rostral brainstem contained cholinergic neurons in the oculomotor, trochlear, and parabigeminal nuclei as well as within the reticular formation. The cholinergic neurons of the reticular formation were the most numerous and formed two intersecting constellations. One of these, designated Ch5, reached its peak density within the compact pedunculopontine nucleus but also extended into the regions through which the superior cerebellar peduncle and central tegmental tract course. The second constellation, designated Ch6, was centered around the laterodorsal tegmental nucleus and spread into the central gray and medial longitudinal fasciculus. There was considerable transmitter-related heterogeneity within the regions containing Ch5 and Ch6. In particular, Ch6 neurons were intermingled with catecholaminergic neurons belonging to the locus coeruleus complex. The lack of confinement within specifiable cytoarchitectonic boundaries and the transmitter heterogeneity justified the transmitter-specific Ch5 and Ch6 nomenclature for these two groups of cholinergic neurons. The cholinergic neurons in the nucleus basalis (Ch4) and those of the Ch5-Ch6 complex were both characterized by perikaryal heteromorphism and isodendritic arborizations. In addition to choline acetyltransferase, the cell bodies in both complexes also had high levels of acetylcholinesterase activity and nonphosphorylated neurofilament protein. However, there were also marked differences in cytochemical signature. For example, the Ch5-Ch6 neurons had high levels of NADPHd activity, whereas Ch4 neurons did not. On the other hand, the Ch4 neurons had high levels of NGF receptor protein, whereas those of Ch5-Ch6 did not. On the basis of animal experiments, it can be assumed that the Ch5 and Ch6 neurons provide the major cholinergic innervation of the human thalamus and that they participate in the neural circuitry of the reticular activating, limbic, and perhaps also extrapyramidal systems. PMID- 2545748 TI - Effect of highly purified eicosapentaenoic acid on psoriasis. PMID- 2545749 TI - Infectious papillomavirus in the vapor of warts treated with carbon dioxide laser or electrocoagulation: detection and protection. AB - Papillomavirus DNA has been reported recently in the vapor (smoke plume) derived from warts treated with carbon dioxide laser; this raises concerns for operator safety. We therefore have studied a group of human and bovine warts to define further the potential risk of wart therapy and to test whether a surgical mask could reduce exposure. Half of each wart was treated with carbon dioxide laser and the other half with electrocoagulation. The vapor produced by each form of therapy was collected with a dry filter vacuum apparatus and analyzed for the presence of papillomavirus. Vapor from human plantar warts was analyzed for the presence of human papillomavirus DNA, because there is no infectivity assay for human papillomavirus. Of plantar warts treated, five of eight laser-derived vapors and four of seven electrocoagulation-derived vapors were positive for human papillomavirus DNA. Greater amounts of papillomavirus DNA were usually recovered in the laser vapor than in the electrocoagulation vapor from the same wart. Bioassay readily detected infectious bovine papillomavirus in the vapor from bovine warts treated with either modality; more virus was present in laser derived material. A surgical mask was found capable of removing virtually all laser- or electrocoagulation-derived virus, strongly suggesting that such masks can protect operators from potential inhalation exposure to papillomavirus. PMID- 2545750 TI - Response of mononuclear leukocyte cyclic adenosine monophosphate phosphodiesterase activity to treatment with topical fluorinated steroid ointment in atopic dermatitis. AB - Our studies of mononuclear leukocyte peripheral blood homogenates demonstrate significantly increased cyclic adenosine monophosphate-specific phosphodiesterase activity in patients with atopic dermatitis who were untreated for 1 week, compared with normal adult nonatopic control subjects. Phosphodiesterase activity is not related to the extent or activity of the patient's disease or the presence or absence of allergic respiratory disease. Enzyme kinetic studies showed a triphasic plot in normal mononuclear leukocytes but a biphasic plot in atopic dermatitis. This may be interpreted as an absence of an enzyme with a low (0.080) Michaelis Menton constant (Km) in atopic dermatitis samples. One week of therapy with a topical fluorinated steroid ointment caused a significant reduction in disease activity. Although a slight reduction in mean total phosphodiesterase activity occurred, it did not reach statistical significance. One week's treatment, however, caused the abnormal biphasic kinetic plot to revert to a triphasic plot with return of the low Km enzyme form in those patients who showed a fall in phosphodiesterase activity. This finding suggests that the elevated phosphodiesterase activity in atopic dermatitis may be responsive in a limited degree to topical steroid therapy. PMID- 2545751 TI - Multiexponential proton spin-spin relaxation in MR imaging of human brain tumors. AB - In vivo measurements of proton relaxation processes in human brain tumors have been performed by magnetic resonance (MR) imaging using a whole-body superconductive MR scanner, operating at 1.5 T. The T1 and T2 relaxation time measurements were based on a combined Carr-Purcell/Carr-Purcell-Meiboom-Gill sequence with two interleaved repetition times and 32 echoes. First, comparative measurements in the imager and with the spectrometer of relaxation times were performed on phantoms containing fluids of different T1 and T2 to evaluate accuracy. A maximum deviation of approximately 10% was found. Multislicing with a gap width of one slice thickness influenced the accuracy of T1 relaxation measurement. A gap width of at least two times the slice thickness was necessary for reliable determination of T1. No influence on T2 values was observed by multislicing. Second, in human head imaging the multiexponential behavior of the T2 decay curves has been analyzed in each pixel, where the mean square deviation has been used as a criterion to discriminate between mono- and biexponential behavior. Mean values of monoexponential T1 and multiexponential T2 relaxation data for white matter, gray matter, CSF, edema, and tumor were sampled in 12 patients with brain tumors. T2 showed monoexponential behavior in white and gray matter, whereas CSF, edema, and tumor showed distinct biexponentiality. The biexponential analysis generally yields "fast" and "slow" components with T2f = 80 +/- 17 ms and T2s = 2,030 +/- 210 ms for CSF (partial volume effect), T2f = 104 +/- 25 ms and T2s = 677 +/- 152 ms for edematous tissues, T2f = 97 +/- 19 ms and T2s = 756 +/- 99 ms for tumor tissues, respectively. Using a stepwise discriminant analysis by forward selection, the two best discriminating parameters of the multiexponential relaxation analysis for each pair of classification groups have been selected. For the discrimination of edematous and tumor tissues a retrospective overall accuracy of 94% has been found. PMID- 2545753 TI - MR features of fleeting CNS lesions associated with Epstein-Barr virus infection. AB - The magnetic resonance and CT findings in a 10-year-old patient with neurologic abnormalities associated with Epstein-Barr virus infection are reported. Multiple focal lesions of various sizes were demonstrated with involvement at one time or another of both cerebral hemispheres, the right thalamus, the right cerebellar hemisphere, and the right optic nerve. A unique finding was the fleeting nature of the lesions with new lesions appearing as others resolved. The clinical presentation, laboratory studies, and radiologic patterns suggest the diagnosis of immune-related acute disseminated encephalomyelitis associated with Epstein Barr virus infection. PMID- 2545752 TI - Malignant cardiac fibrous histiocytomas and angiosarcomas: MR features. AB - Primary tumors of the heart are rare, and one-third of them are malignant. The magnetic resonance (MR) features, their specificity, and significance in two patients with malignant fibrous histiocytoma and three patients with angiosarcoma are presented and the literature briefly reviewed. Malignant fibrous histiocytoma occurred in the left atrium and demonstrated slightly heterogeneous intermediate signal intensity on T1-weighted images and high signal intensity on T2-weighted images. The angiosarcomas involving primarily the right heart were more varied in MR appearance with heterogeneous signals. The extensive angiosarcomas almost circumferentially involving the epicardium and pericardium showed the "cauliflower" appearance with focal areas of increased signal intensity probably related to thrombosis or hemorrhage. Magnetic resonance provided detailed anatomic information and characterization of malignant cardiac tumors. PMID- 2545755 TI - High amounts of soyhulls for pelleted concentrate diets. AB - Twelve multiparous Holstein cows were used in a balanced, two-period, changeover design to evaluate soyhulls as a replacement for corn grain in lactation diets. Soyhulls constituted 0 (corn), 50 (corn-soyhull), and 95% (soyhulls) of the concentrate mixture, which was prepared as a 4.8-mm pellet. These concentrate mixtures were mixed with alfalfa silage (50:50 DM) and fed ad libitum as complete diets. Mean DM intakes averaged 23.7 kg/d and were similar for corn, corn soyhulls, and soyhull diets. Corresponding daily milk yields averaged 29.8, 28.9, and 27.3 kg and milk fat averaged 3.13, 3.33, and 3.49%, resulting in similar 3.5% FCM yields and feed efficiency. Dry matter digestibility was higher for the corn diet (70%) and the corn-soyhull diet (69%) compared with the soyhull diet (61%). Digestibilities (%) of NDF were 55, 63, and 58; ADF were 56, 62, and 55; and CP were 66, 64, and 59 for corn, corn-soyhull, and soyhull diets, respectively. The gastrointestinal passage rate of alfalfa and of soyhulls was similar among diets. The energy value of soyhulls appears to be equal to corn in pelleted concentrates. The fat-depressing effect of pelleting a high starch (corn) concentrate was minimized by including soyhulls. PMID- 2545757 TI - Multiple glomus tumors: a role for magnetic resonance imaging in patient evaluation? AB - Glomus tumors may occur as solitary lesions, or rarely as multiple glomus tumors (MGT). Internal organ involvement in patients with MGT has been reported, but is rare. Magnetic resonance imaging (MRI) has been reported to be useful in evaluating patients with blue rubber bleb nevus syndrome for nongastrointestinal internal organ involvement. Three patients with MGT were evaluated by MRI for both cutaneous and internal involvement. MRI, which is useful in evaluating other vascular lesions, is not useful in the evaluation of patients with multiple glomus tumors. With these results, patients with MGT can be spared the expense of MRI procedure. PMID- 2545754 TI - The potential of a phyllosilicate (Palabora vermiculite) as buffer in dairy cattle diets. AB - Four rumen-fistulated Holstein cows were used to determine the ability of vermiculite to alter rumen fermentation, rumen fluid dilution rate, milk production, and nutrient utilization in a 4 x 4 Latin square experiment. Treatments consisted of 1) basal diet (70% concentrate:30% Eragrostis curvula hay), 2) basal + .6% NaHCO3 + 1.8% vermiculite, 3) basal + 1.2% NaHCO3, and 4) basal + 3.6% vermiculite. Feed intake and milk production were not affected, but the NaHCO3 treatments tended to increase milk fat production. The 1.2% NaHCO3 treatment increased rumen pH and fluid dilution rate, decreased molar percent propionate, and increased acetate:propionate ratio. Rumen NH3 N was not affected. Milk yield (kg/d) and milk fat (%) for the treatments were 1) 19.3, 3.33; 2) 19.4, 3.59; 3) 19.8, 3.62; and 4) 18.7, 3.32. Rumen fluid pH, dilution rate (%/h), and acetate:propionate ratio were 1) 5.75, 10.9, 2.08; 2) 5.89, 11.5, 2.16; 3) 5.95, 12.1, 2.38; and 4) 5.80, 11.6 and 1.82. Treatments had little effect on nutrient digestibilities and serum mineral concentrations. Vermiculite did not show any promise as a buffer but NaHCO3 proved to be an effective buffer in diets based on Eragrostis hay and 70% concentrate diets. PMID- 2545756 TI - Influence of protein supplementation of alkaline hydrogen peroxide-treated wheat straw on ruminal microbial fermentation. AB - A dual flow continuous culture system was used to determine the effects of four protein sources (soybean meal, Ca-ligno-sulfonate treated-soybean meal, blood meal, and feather meal), supplied individually or in combination in diets composed predominantly of alkaline hydrogen peroxide-treated wheat straw, on ruminal microbial fermentation and amino acid flow. Diets containing blood meal had lower organic matter and fiber digestion, NH3 N and VFA concentrations, and CP degradation but higher non-NH3 N, dietary N, total amino acid, and essential amino acid flows. Feather meal fed alone or combined with other sources was not different from the blood meal diet in organic matter and fiber digestion. Combining treated soybean meal and blood meal resulted in similar organic matter and fiber digestibilities compared with the soybean meal diets. This combination was similar to the treated soybean meal diet in VFA concentration, non-NH3 N, and essential and total amino acid flows; however, amino acid profile was different with this combination, showing higher histidine and leucine flows. Results from this study suggest that amino acid profiles of digesta leaving the rumen may be manipulated by choice of protein supplement when diets containing a low protein, highly fermentable feedstuff such as alkaline hydrogen, peroxide-treated wheat straw are fed. PMID- 2545758 TI - [Effect of the hyperbaric oxygenation of animals and man on mitochondrial function in their tissues (based on EPR study data)]. AB - The ration of differential intensities of EPR signal of free endogenous radicals of semiquinone type and iron-sulfur proteins recorded at the temperature of liquid nitrogen in tissues (R index) is proposed as a criterion for estimation of changes in the functional state of mitochondria in human and animal tissues treated with hyperbaric oxygenation (HBO). The increase in R value was observed in hearts of intact mice and rabbits treated with HBO in near-toxic doses. This indicates a shift in mitochondria redox state towards oxidation. The above effect was not observed in mitochondria from intact zone of myocardium in animals with experimental infarction, and the numbers of mitochondria in the tissue increased after HBO. HBO treatment of women with pathological pregnancy leads to the decrease in R value for placental mitochondria. PMID- 2545759 TI - [Effect of hydrazine derivatives, plant growth regulators, on the cytochrome oxidase reaction]. AB - Gidrel, digidrel, and sodium salt of maleic hydrazide were shown to exert no influence on oxygen consumption by cytochrome oxidase in concentrations up to 10( 1) M. Similar concentrations of N,N-dimethylamino-succinic acid hydrazide inhibited cytochrome oxidase but did not influence the optical spectrum of cytochrome oxidase. PMID- 2545761 TI - A stopped-flow mixer device for a batch microcalorimeter application to NAD NADase reaction. AB - A new molded polypropylene, diamond-like carbon (DLC)-coated mixing cell has been developed for use in the batch microcalorimeter. Reagent volume can be varied from 25 microliters to 100 microliters. A 10 microcalorie reaction heat can be measured to 5%. Repeat reactions can be done as often as every 10 min for a fast reaction. Reactions can be started within 1 h or less after loading. A pre equilibrator and a temperature-controlled syringe drive unit permit solutions to be stored at 4 degrees C while being run at any temperature from -20 degrees C to 40 degrees C. The kinetics and enthalpy of reaction of NAD-NADase have been measured. delta H is about 21 kcal/mol endothermic. PMID- 2545760 TI - [Photogeneration of the superoxide anion oxygen radical and its role in the inactivation of yeast cells by long-wave UV light]. AB - Long-wave (320-400 nm) UV-induced oxygen superoxide anion radical (O2-) formation was found in yeast cells. This radical plays an important part in initiation of photodestruction reactions in DNA which serves as a main target of UV irradiation in yeast. The observed cell photoinactivation spectrum at the wavelengths 320-400 nm suggests that NADH can serve as an endogenous sensitizer of O2- formation. PMID- 2545762 TI - Spin trapping of the superoxide anion: complications in the use of the water soluble nitroso-aromatic reagent DBNBS. AB - Sodium 3,5-dibromo-4-nitrosobenzenesulfonate (DBNBS) is reported to be a useful spin trap for the measurement of superoxide anions in aqueous solution. However, the signal observed arises from interaction of the spin trap with some species other than the superoxide radical or hydrogen peroxide, a product of its dismutation, as the addition of both superoxide dismutase and catalase to a superoxide generating system failed to attenuate the signal. Therefore caution must be employed in the interpretation of results obtained using this spin trap. PMID- 2545763 TI - Lack of aldosterone inhibition by atrial natriuretic factor in primary aldosteronism: in vitro studies. AB - Several studies demonstrated that aldosterone secretion, by bovine, rat and human glomerulosa cells, is inhibited in vitro by atrial natriuretic factor (ANF). This effect has also been investigated with conflicting results in cells taken from aldosterone-producing tumors. In the present study, atrial natriuretic factor has been tested on aldosteronoma cells obtained from 4 patients with primary aldosteronism. The cells were studied both with perfusion and incubation systems. Aldosterone secretion was stimulated by ACTH, angiotensin II and potassium with or without ANF 10 microM. In this study ANF lacked to inhibit either basal and stimulated aldosterone secretion, indicating some alterations of ANF-adrenal interaction in this syndrome. PMID- 2545764 TI - Pituitary-adrenal responses to CRH and insulin hypoglycemia in patients with idiopathic GH deficiency. AB - It is well known that the activation of the hypothalamus-pituitary-adrenal axis (HPA) by insulin-induced hypoglycemia (IIH) is more potent and multifactorial than that caused by CRH administration. In this study we compared the clinical value of both tests in assessing the integrity of the HPA system. Plasma ACTH and cortisol responses to oCRH (1 microgram/kg iv) and IIH (insulin 0.1 U/kg iv, glycemia less than 40 mg/dl) were compared in 15 patients with idiopathic GH deficiency. The CRH-induced mean ACTH response was lower, but not significantly, in patients than in controls (peak: 8.8 +/- 1.7 vs 13.4 +/- 2.2 pmol/l), while the mean cortisol response was significantly lower than in normals (peak: 585.7 +/- 49.5 vs 764.5 +/- 52.2 nmol/l, p less than 0.005). Plasma ACTH and cortisol responses to IIH were significantly lower than in normal subjects (peak: 22.3 +/- 5.3 vs 35.8 +/- 5.2 pmol/l, p less than 0.05 and peak: 566.5 +/- 55 vs 803 +/- 38.5 nmol/l, p less than 0.02, respectively). Both in controls and in patients the CRH-induced mean ACTH response was significantly lower (p less than 0.02) than that after insulin, while cortisol peaks were not different. In conclusion, in patients with GH deficiency the impairment of ACTH secretion is not evident in basal condition, but it is disclosed after appropriate dynamic tests. It is confirmed that insulin hypoglycemia is a more potent stimulus than CRH for ACTH release. PMID- 2545765 TI - Calcitonin suppresses growth hormone (GH) response to growth hormone-releasing hormone (GHRH) in man. AB - Calcitonin (CT) receptors have been found in the hypothalamus, suggesting a neuroendocrine role for this peptide. We have recently shown that, in the rat, central administration of salmon calcitonin (sCT) suppresses basal and GHRH stimulated GH secretion. To further investigate how sCT alters GH secretion, we studied the effects of sCT (100U MRC, im) or placebo on basal and GHRH (50 micrograms, iv)-stimulated GH secretion in 6 normal men. GHRH was administered 1 h after sCT injection. Basal GH levels were not altered by sCT administration. However, GH response to GHRH was markedly suppressed by sCT (area under the curve - sCT: 574.6 +/- 69.7 vs placebo: 1057.2 +/- 284.8 micrograms. min/L; p less than 0.02). Cortisol levels were higher in sCT-treated subjects compared to controls, from 45 to 105 min after sCT injection (p less than 0.05). However, no correlation was found between GH response to GHRH and cortisol levels. No changes in glucose, calcium and PTH levels were seen. These results demonstrate that sCT inhibits GHRH-induced GH secretion in man by a mechanism apparently independent of changes in peripheral cortisol, glucose, calcium and PTH levels. PMID- 2545766 TI - Effects of the opiate agonist loperamide on pituitary-adrenal function in patients with suspected hypercortisolism. AB - In the present work the possible use of loperamide, an opiate agonist, in the dynamic evaluation of patients with suspected hypercortisolism was investigated. The effects of loperamide on plasma ACTH and cortisol levels were evaluated in normal subjects and in 58 patients with suspected Cushing's syndrome. The results were compared to those obtained after the overnight dexamethasone suppression test. In normal subjects plasma ACTH and cortisol levels were significantly (p less than 0.005) suppressed by both loperamide (16 mg po) and dexamethasone (1 mg po). In 17 patients, in whom the diagnosis of Cushing's syndrome was confirmed by subsequent investigations, neither loperamide or dexamethasone inhibited cortisol (from a baseline of 606 +/- 55 nmol/L) to a nadir of 502 +/- 43 nmol/L and 539 +/ 50 nmol/L, respectively) and ACTH concentration (from a basal level of 70.1 +/- 11.8 pg/ml to a nadir of 46.0 +/- 8.6 pg/ml and 54.3 +/- 7.5 pg/ml, respectively). In 34 patients, in whom the suspect of hypercortisolism was ruled out, either loperamide or dexamethasone suppressed the pituitary-adrenal axis: cortisol and ACTH levels significantly fell from 417 +/- 24 nmol/L and 28.3 +/- 3.5 pg/ml to 60 +/- 6 nmol/L and 14.4 +/- 1.4 pg/ml after loperamide and to 26 +/ 4 nmol and 16.4 +/- 1.7 pg/ml after dexamethasone. In 7 patients discordant responses were observed. In 3 patients treated with antiepileptic drugs ACTH and cortisol levels were inhibited by loperamide, but not by dexamethasone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545767 TI - Effects of bromocriptine on pituitary and adrenal cortex in pre-adrenarchal rabbits. AB - The effect of bromocriptine on the morphological picture and steroid content of the adrenal gland, and on certain pro-opiomelanocortin (C) peptides in the pituitary gland was evaluated in preadrenarchal rabbits. Eighteen immature male rabbits (5 weeks of age), were treated for 10 days with saline (n = 10,2 ml sc) or bromocriptine mesylate (n = 8, 3 mg/kg sc) two times/day. After the last administration all animals received dexamethasone (0.25 mg im) and the next morning, 60 min after ACTH injection (0.25 mg im), plasma was drawn and they were sacrificed. Adrenals and pituitaries were immediately removed. For each animal, one adrenal gland was fixed, dehydrated and embedded in paraffin for histology; the other one was stored in saline for determination of androstenedione (A), dehydroepiandrosterone (DHA), 17-OH progesterone (17 P), and cortisol. Steroids were analyzed by RIA after previous extraction and celite-ethyleneglycol chromatography, or directly (cortisol). The immunoreactivities (ir) related to beta-Endorphin (B-EP), ACTH and alpha-MSH were evaluated in pituitary homogenates using specific RIAs. The bromocriptine-treated rabbits showed a significant increase in the percentage of the adrenal zona reticularis (21.5 +/- 3.9% of total cortex vs. 12.7 +/- 1.3% in controls, p less than 0.05, mean +/- SE), and a decrease of the zona fasciculata (57.6 +/- 3.13% vs. 67.7 +/- 2.05% in controls, p less than 0.05). No significant changes were observed in the relative percentage of the zona glomerulosa.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545768 TI - Bilateral and simultaneous venous sampling of inferior petrosal sinuses for ACTH and PRL determination: preoperative localization of ACTH-secreting microadenomas. AB - We report our experience on venous sampling of the inferior petrosal sinuses for basal and CRH-stimulated ACTH and PRL gradients in 8 patients with surgically proven Cushing's disease who had normal preoperative neuroradiological studies. In 7 patients basal plasma ACTH concentrations in the inferior petrosal sinus ipsilateral to the tumor were higher than in the contralateral sinus; the gradients were enhanced by oCRH administration. In one out of two patients who had previously undergone unsuccessful pituitary microsurgery, neither basal nor oCRH-induced ACTH increases led to correct localization of the microadenoma within the pituitary. In 4 out of 7 patients basal serum PRL concentrations in the inferior petrosal sinus ipsilateral to the tumor were higher than in the contralateral; only two out of 4 showed an increase in PRL levels after oCRH injection. Our study confirms that simultaneous and bilateral venous sampling of inferior petrosal sinuses is a valuable means to identify the site of microadenomas in patients with Cushing's disease without neuroradiological evidence of the tumor. This procedure may give misleading results in patients previously operated on. Unilateral or predominant increases of PRL concentration during catheterization of the inferior petrosal sinuses, when present, always lateralize to the side of the corticotroph adenoma, providing a possible additional signal of the presence of the tumor. PMID- 2545769 TI - Idiopathic isolated ACTH deficiency and the response to CRF. AB - A 26-year-old woman, intermittently symptomatic over 8 yr, was evaluated for weakness, lightheadedness, and gastrointestinal complaints. She was found to have negligible plasma cortisol and low to low-normal plasma ACTH levels. Although serum cortisol was responsive to ACTH stimulation, metyrapone and insulin-induced hypoglycemia were unable to stimulate pituitary driven adrenal activity. Hypothalamic and pituitary functions were otherwise intact. CRF administration failed to stimulate any increase in plasma ACTH or cortisol levels and CT scanning failed to reveal any anatomic abnormality of the sella or suprasellar areas consistent with an acquired isolated defect in pituitary ACTH secretion. These studies support the impression that the primary defect in isolated idiopathic ACTH deficiency is failure of pituitary ACTH-secreting cells. PMID- 2545772 TI - Infection control measures for human parvovirus B19 in the hospital setting. PMID- 2545771 TI - Multiple sclerosis and brain tumor: a diagnostic challenge. AB - Multiple sclerosis may present as a lesion indistinguishable clinically and radiologically from brain tumor. A case of multiple sclerosis is reported with clinical and radiologic features of a brain tumor. A brain biopsy was later undertaken that provided pathological confirmation of the diagnosis of this demyelinating disease. This case report in conjunction with those in the literature indicate that conservative treatment with steroid therapy and serial computed tomography should establish the diagnosis of multiple sclerosis without the need for surgical intervention. PMID- 2545773 TI - The role of dopaminergic and noradrenergic receptors in human TSH and LH release. AB - The present study was undertaken to evaluate the relative roles of noradrenergic (NA) and dopaminergic (DA) neurotransmission in the control of thyrotropin (TSH) and luteinizing hormone (LH) release. Oral dextroamphetamine 20 mg caused a rise in the serum levels of TSH and LH in 24 healthy male volunteer subjects. The increase in TSH secretion was augmented by prior treatment with pimozide and reduced by thymoxamine. Conversely the LH response to destroamphetamine was reduced by pimozide and possibly accentuated by thymoxamine. The results confirm an inhibitory role for DA receptors in the control of TSH release and indicate that NA receptors may exert a facilitatory role. The converse would appear to be the case with LH, with DA being facilitatory and NA possibly inhibitory. PMID- 2545770 TI - Somatomedins: chemical and functional characteristics of the different molecular forms. PMID- 2545775 TI - Mutation of 3' splice sites in two different class I genes results in different usage of cryptic splice sites. AB - To determine the pattern of alternative splicing at the 5' end of class I genes, the 3' splice sites bordering exon 2 of the H-2Dd and H-2Kd genes were mutated from AG to GG (H-2Dd) or CG (H-2Kd). The mutant genes were transfected into L cells, and RNA from clones expressing these Ag was used for analysis by RNase and S1 nuclease mapping techniques. The first intervening sequence of both class I genes contains several potential 3' splice acceptor sites. However, a clear preference for only one site was detected in each of the H-2Dd and H-2Kd mRNA. Examination of the endogenous H-Dd and H-2Kd class I transcripts in normal murine tissues and in tumors demonstrated that the alternatively spliced mRNAs were produced, but at a low frequency. Infection of transfected L cells or tumor lines with vesicular stomatitis virus altered the level of differentially spliced message in these cells. PMID- 2545774 TI - Laminar pattern of cholinergic and adrenergic receptors in rat visual cortex using quantitative receptor autoradiography. AB - The laminar distribution of muscarinic acetylcholine receptors, including the M1 receptor subtype, of beta-adrenergic receptors, and noradrenaline uptake sites, was studied in the adult rat visual, frontal, somatosensory and motor cortex, using quantitative receptor autoradiography. In the visual cortex, the highest density of muscarinic acetylcholine receptors was found in layer I. From layer II/III to layer V binding decreases continuously reaching a constant binding level in layers V and VI. This laminar pattern of muscarinic receptor density differs somewhat from that observed in the non-visual cortical regions examined: layer II/III contained the highest receptor density followed by layer I and IV: lowest density was found in layer V and VI. The binding profile of the muscarinic cholinergic M1-subtype through the visual cortex shows a peak in cortical layer II and in the upper part of layer VI, whereas in the non-visual cortical regions cited the binding level was high in layer II/III, moderate in layer I and IV, and low in layer VI. Layers I to IV of the visual cortex contained the highest beta adrenergic receptor densities, whereas only low binding levels were observed in the deeper layers. A similar laminar distribution was found also in the frontal, somatosensory and motor cortex. The density of noradrenaline uptake sites was high in all layers of the cortical regions studied, but with noradrenaline uptake sites somewhat more concentrated in the superficial layers than in deeper ones. The distinct laminar pattern of cholinergic and noradrenergic receptor sites indicates a different role for acetylcholine and noradrenaline in the functional anatomy of the cerebral cortex, and in particular, the visual cortex. PMID- 2545776 TI - Relationship between target cell recognition and temporal fluctuations in intracellular Ca2+ of human NK cells. AB - Temporal changes in intracellular Ca2+ concentration, [Ca2+]i, of resting human peripheral blood NK cells in response to target cell binding were evaluated by flow cytometry. [Ca2+]i was significantly elevated in PBL and purified NK cells bound to NK-sensitive K562 and HSB2 target cells, but not in those bound to NK resistant MD1 B-lymphoblastoid cells. Thus, a) the ability of a target cell to elicit a Ca2+ flux response correlated with its sensitivity to lysis of NK cells, and b) adhesion alone was not a sufficient stimulus for response induction. Conjugates of NK cells bound to K562 target cells were sorted onto agarose-coated slides on the basis of relative NK cell [Ca2+]i and evaluated in 19-hr single cell agarose cytotoxicity assays. In contrast to those with basal levels of [Ca2+]i, NK cells with elevated [Ca2+]i bound more strongly to target cells, as judged by the stability of conjugates to sort-related shear forces (p less than 0.01), and more frequently killed the target cell to which they were attached (p less than 0.05). Temporal fluctuations in [Ca2+]i were observed in target-bound NK cells in both the presence and absence of extracellular Ca2+. Thus, influx of extracellular Ca2+ and release of Ca2+ from internal stores both appeared to contribute to the NK cell Ca2+ flux response triggered by adhesion to appropriate target cells. These results support the hypothesis that such fluctuations in NK cell [Ca2+]i constitute an early signal flagging the occurrence of NK cell recognition. PMID- 2545778 TI - Platelet-derived growth factor and IL-1 interactions in rheumatoid arthritis. Regulation of synoviocyte proliferation, prostaglandin production, and collagenase transcription. AB - We show that platelet-derived growth factor (PDGF) and IL-1 interact in both a synergistic and antagonistic manner to regulate synovial fibroblast-like cells (synoviocytes) derived from patients with rheumatoid arthritis. PDGF and IL-1 operated synergistically in vitro to stimulate synoviocyte proliferation in the presence of indomethacin. However, when these same cells were treated with PDGF and IL-1 in the absence of indomethacin, IL-1 inhibited PDGF-stimulated synoviocyte proliferation. Moreover, exogenous PGE2, a PG known to be produced in response to IL-1, dramatically inhibited synoviocyte proliferation induced by PDGF. PDGF also acted synergistically to markedly increase production of PGE2 stimulated by IL-1. This is in contrast to the antagonistic effect PDGF had on IL 1-stimulated collagenase transcription. IL-1 stimulated collagenase transcription, but PDGF did not. It in fact inhibited IL-1 stimulation of collagenase gene expression. These data differ somewhat from those reported for dermal fibroblasts. Our data further indicate that the effects of cytokines vary from one cell type to another, even amongst "fibroblasts," and illustrate the complexity of cytokine regulation of rheumatoid synoviocyte function. PMID- 2545777 TI - Stimulation of PIP2 hydrolysis by aluminum fluoride in resting T cell subsets of normal and autoimmune-prone lpr mice. AB - T cells of autoimmune-prone mice homozygous for the lpr mutation respond poorly to mitogens in terms of proliferation and of IL-2 production. In a previous study, we have correlated this deficient activation with the inability of mitogens to stimulate hydrolysis of phosphatidylinositol 4,5-bisphosphate in lpr T cells, although these cells bind mitogen and express the TCR/CD3 complex. In order to determine whether activation-deficient lpr T cells contain functional GTP-binding (G) protein(s) and phospholipase C, we examined the effects of the G protein activating agent sodium fluoride plus Al+3 (AlF-4). AlF-4 stimulated phosphatidylinositol turnover, a response characteristic of TCR/CD3 occupancy, in mature L3T4+ and Ly2+ T cells. Second, and more important, AlF-4 stimulated the same biochemical events in L3T4-, Ly2- (double-negative) T cells from the normal thymus or from the enlarged lymph nodes of autoimmune-prone mice homozygous for the lpr mutation. However, these double-negative T cells were unresponsive to receptor-active ligands such as T cell mitogens or anti-CD3-epsilon mAb, despite their ability to bind these ligands. These findings suggest that activation deficient double negative T cells express the receptors, G protein(s) and effector enzymes necessary for second messenger formation and further suggest that the failure of these cells to generate the relevant second messengers in response to mitogens or anti-CD3-epsilon antibody may be due to inefficient coupling of the TCR/CD3 complex to G proteins. PMID- 2545779 TI - Functional and phenotypic alterations in T cell subsets during the course of MAIDS, a murine retrovirus-induced immunodeficiency syndrome. AB - The functional and phenotypic characteristics of Ly-4(CD4)+ and Ly-2(CD8)+ T cells were studied after induction of murine AIDS with LP-BM5 murine leukemia virus. Assays of spleen cells for their ability to generate in vitro CTL responses to TNP-modified autologous cells (self + x CTL) and to alloantigens (allo CTL) showed that self + x CTL responses were greatly impaired at 3 to 4 wk postinfection and were undetectable thereafter. Allo CTL responses were normal at 3 to 4 wk, but were reduced at 8 to 9 wk and absent at 14 wk postinfection. This sequential loss of self + x and allo CTL responses was related to a selective defect in Ly-4(CD4)+ Th cell function associated with impaired production of IL-2 and deficient proliferative responses to Con A or to soluble Ag. Changes in the functional characteristics of Ly-4(CD4)+ T cells were unrelated to changes in their frequency in spleen, but did correlate with marked alterations in their distribution among four subsets defined by mAb SM3C11 and SM6C10. Assays of CTL responses generated by mixtures of spleen cells from normal and infected mice suggested that active suppression of Ly-4(CD4)+ Th function may contribute to this defect. Studies of Ly-2(CD8)+ T cells showed that infection with LP-BM5 murine leukemia virus also induced a major phenotypic shift in subpopulations defined by their reactivity with mAb 6C10. However, this phenotypic change did not appear to correlate with major functional defects. PMID- 2545780 TI - Platelet-activating factor (PAF) enhances tumor necrosis factor production by alveolar macrophages. Prevention by PAF receptor antagonists and lipoxygenase inhibitors. AB - The effect of platelet-activating factor (PAF) on TNF production by rat alveolar macrophages (AM) and the role of endogenous leukotriene B4 (LTB4) in this regulation were examined. When AM were cultured with PAF alone, no change in TNF production was observed. However, the concomitant addition of PAF and muramyl dipeptide to AM cultures markedly enhanced (2- to 3-fold) TNF production in a concentration-dependent fashion with peak effect at 10(-10)M PAF. This enhancement occurred when muramyl dipeptide and PAF were present together at the initiation of the 24-h culture. Stimulation of TNF production by PAF was blocked by specific, but structurally different PAF receptor antagonists, BN 52021, CV3988 and WEB 2086. Additionally, the stereoisomer of PAF, [S]PAF, and the biologically inactive precursor/metabolite of PAF, lyso-PAF failed to induce significant enhancement in TNF production. In parallel, addition of PAF to AM triggered LTB4 release in a concentration-dependent manner. Inhibition of 5 lipoxygenase by nordihydro-guaiaretic acid or AA-861 blocked the PAF-induced augmentation of both TNF and LTB4 production. This was partially reversed by addition of exogenous LTB4. Collectively, these data suggest that PAF enhances TNF production by interaction with a specific putative receptor and by subsequent induction of endogenous 5-lipoxygenase activity in AM. PMID- 2545781 TI - Characterization of B lymphocytes present in the demyelinating lesions induced by Theiler's virus. AB - Theiler's virus, a murine picornavirus, persists in the central nervous system of SJL/J mice and causes inflammation and demyelination in the white matter of spinal cord. We isolated inflammatory cells from the central nervous system of infected animals and studied their functions in vitro. Flow microfluorimetry analysis showed the presence of all major lymphocyte subsets, namely CD4+ and CD8+ T cells as well as B lymphocytes. B lymphocytes were activated in vitro and the antigenic specificity of secreted Ig was determined by immunoblotting. Secreted Ig reacted strongly with viral capsid proteins VP1 and VP2 and had neutralizing activity. They reacted also with two nonviral white matter components which were present only in infected animals. Therefore, it is likely that Igs secreted at the site of infection play a role in limiting virus spread. It is also possible that virus induced autoreactive antibodies participate in demyelination. PMID- 2545783 TI - The release of C5a in complement-activated serum does not require C6. AB - The influence of terminal complement components on the generation and release of the complement C5a fragment was investigated by comparing the levels of C5a in complement-activated serum with the levels of C5a produced in serum depleted of complement C6. In order to investigate the release of C5a, a modified C5a assay was developed that utilizes an anti-C5b monoclonal antibody to remove C5, C5b, and C5b-C5a complexes from samples prior to C5a assay. The modified assay was developed because the standard methodology, which includes an acid-precipitation step designed to dissociate C5a and C5b, cannot distinguish free C5a from the C5a that is bound to C5b. Therefore, the standard methodology is not capable of monitoring the influence of terminal components on C5a/C5b dissociation. Levels of C5a were measured in complement-activated whole human serum, in serum depleted of C6, and in serum containing inhibitory levels of anti-C6 Fab using both the modified C5a assay and the standard methodology. Sera were complement-activated with either zymosan to activate the alternative complement pathway or with antibody-coated sheep erythrocytes to activate the classical pathway. The levels of free C5a in C6-depleted sera after activation were equivalent to the C5a levels in activated whole serum, indicating that C6 is not required for the release of C5a from C5b. In addition, the quantity of C5a detected in zymosan activated sera using the standard acid-precipitation methodology was greater than C5a levels when assayed using the modified immunoadsorption technique, confirming that acid-treatment enhances the C5a dissociation and promotes C5a recovery. Since the other terminal components, C7, C8, and C9, bind to C5b only after C5b only after C6 is bound, these results indicate that none of the terminal components are required for the release of C5a. Although the terminal components could influence the rate of C5a release, the quantity of C5a released in serum was entirely independent of terminal components. PMID- 2545782 TI - Dextran sulfate and heparin interact with CD4 molecules to inhibit the binding of coat protein (gp120) of HIV. AB - Dextran sulfate, heparin, and certain other sulfated polysaccharides potently inhibit the adsorption of HIV to CD4+ cells. The mechanism of this inhibition is unclear and, specifically, it is unknown if these agents act at the level of CD4 gp120 binding. For example, previous reports have demonstrated that dextran sulfate does not inhibit the cell surface binding of anti-CD4 mAb known to be directed at the gp120 binding site. In order to confirm and extend these observations, in the present study, it was shown that dextran sulfate does not inhibit the binding of OKT4A, OKT4C, Leu3a, or B66.6 to CD4+ cells as measured by cytofluorography. Next, recombinant forms of CD4 (rT4) and gp120 (rgp120) were utilized to directly study their molecular interaction in the absence of other viral or cellular structures. Reciprocal solid phase ELISA assays were developed to study directly the effects of sulfated polysaccharides on the binding of rT4 to immobilized rgp120 and vice versa. Dextran sulfate, heparin, and fucoidan, but not chondroitin sulfate, inhibited the binding of rgp120 to rT4. Importantly, dextran sulfate and heparin pre-treatment of immobilized rT4, but not immobilized rgp120, inhibited rT4-rgp120 binding. Taken together, these data suggest that while both sulfated polysaccharides and anti-CD4 mAb inhibit gp120 binding, the sulfated polysaccharides interact with sites on CD4 that are distinct from those with which the antibodies bind. PMID- 2545784 TI - Different membrane anchors of Fc gamma RIII (CD16) on K/NK-lymphocytes and neutrophils. Protein- vs lipid-anchor. AB - Fc gamma RIII (CD16), the type three receptor for the Fc portion of IgG, is expressed on neutrophils, killer (K)/NK lymphocytes and macrophages. K/NK lymphocyte Fc gamma RIII, which plays a role in antibody-dependent cellular cytotoxicity, is an efficient signal transducing molecule, whereas neutrophil Fc gamma RIII, which plays a role in immune-complex clearance, seems less efficient in signal transduction. Neutrophil Fc gamma RIII has been reported to be a glycan phosphatidylinositol-anchored membrane protein. Our studies suggest that K/NK lymphocyte Fc gamma RIII is protein-anchored rather than glycan phosphatidylinositol-anchored. That is, K/NK lymphocyte Fc gamma RIII was resistant to phosphatidylinositol-specific phospholipase C and surface expression of Fc gamma RIII was not affected on K/NK lymphocytes from patients with paroxysmal nocturnal hemoglobinuria, a disorder of hemopoietic stem cells resulting in deficient expression of glycan-phosphatidylinositol-anchored proteins. Different membrane anchoring mechanisms of the Fc gamma RIII may account for different consequences of the ligand binding to two cell types. PMID- 2545786 TI - Mechanism of arachidonic acid liberation in platelet-activating factor-stimulated human polymorphonuclear neutrophils. AB - Upon stimulation of human polymorphonuclear neutrophils with platelet-activating factor (PAF), arachidonic acid (AA) is released from membrane phospholipids. The mechanism for AA liberation, a key step in the synthesis of biologically active eicosanoids, was investigated. PAF was found to elicit an increase in the cytoplasmic level of free Ca2+ as monitored by fluorescent indicator fura 2. When [3H] AA-labeled neutrophils were exposed to PAF, the enhanced release of AA was observed with a concomitant decrease of radioactivity in phosphatidylinositol and phosphatidylcholine fractions. The inhibitors of phospholipase A2, mepacrine and 2-(p-amylcinnamoyl)-amino-4-chlorobenzoic acid, effectively suppressed the liberation of [3H]AA from phospholipids, indicating that liberation of AA is mainly catalyzed by the action of phospholipase A2. The extracellular Ca2+ is not required for AA release. However, intracellular Ca2+ antagonists, TMB-8 and high dose of quin 2/AM drastically reduced the liberation of AA induced by PAF, indicating that Ca2+ is an essential factor for phospholipase A2 activation. PAF raised the fluorescence of fura 2 at concentrations as low as 8 pM which reached a maximal level about 8 nM, whereas more than nM order concentrations of PAF was required for the detectable release of [3H]AA. Pretreatment of neutrophils with pertussis toxin resulted in complete abolition of AA liberation in response to PAF. However, the fura 2 response to PAF was not effectively inhibited by toxin treatment. In human neutrophil homogenate and membrane preparations, guanosine 5' O-(thiotriphosphate) stimulated AA release and potentiated the action of PAF. Guanosine 5'-O-(thiodiphosphate) inhibited the effects of guanosine 5'-O (thiotriphosphate). These results suggest several points: 1) PAF stimulates human polymorphonuclear neutrophils to liberate AA mainly by the action of phospholipase A2; 2) Ca2+ mobilization alone is not sufficient to stimulate AA release, although Ca2+ is the important factor for phospholipase A2 activation; and 3) a pertussis toxin-sensitive GTP-binding protein may be implicated in activation of phospholipase A2. PMID- 2545785 TI - Mechanisms of T cell activation by the calcium ionophore ionomycin. AB - We have investigated signaling mechanisms that may underlie the T cell mitogenic properties of the Ca2+ ionophore ionomycin. Ionomycin induces highly purified resting human T cells to proliferate in the presence of monocytes with accompanying IL-2R expression and IL-2 synthesis. Treatment of T cells with ionomycin triggers the hydrolysis of phosphoinositides, as evidenced by the accumulation of the hydrolytic by-products phosphatidic acid and inositol phosphates. Ionomycin also induces the activation of protein kinase C (PKC), as demonstrated by the auto-phosphorylation of PKC and the phosphorylation of the PKC target proteins CD4 and CD8. Ionomycin synergizes with PMA in enhancing the activation of PKC. It is concluded that, in addition to its putative activation of Ca2+/calmodulin-dependent signaling pathways, ionomycin induces the hydrolysis of phosphoinositides and the activation of PKC in human T cells. The synergy of ionomycin with phorbol esters in triggering T cell activation may relate, at least in part, to enhanced activation of PKC. PMID- 2545788 TI - Human papillomavirus type 31 infection of the uterine cervix in England. AB - A total of 452 uterine cervical scrapes, from two centres in England, has been examined by dot blot hybridisation for the presence of human papilloma virus (HPV) types 6, 11, 16, 18 and 31. HPV DNA was found in 36.5% samples. Twenty-nine of the women were infected with more than one type. Of those with detectable HPV, 14.5% had HPV type 31 infections. This type appears to cause significant infection of the genital tract in England. PMID- 2545787 TI - Prostaglandin E precursor fatty acids inhibit human IL-2 production by a prostaglandin E-independent mechanism. AB - Essential fatty acids, from which PG derive, can participate in development and regulation of immune responses and have been shown to suppress inflammation and tissue injury in animal models. In this report, we investigate the effects of the immediate (DGLA, precursor to PGE1), arachidonic acid (AA, PGE precursors, dihomogamma linolenic acid (DGLA, precursor to PGE1), arachidonic acid (AA, precursor to PGE2), and eicosapentaenoic acid (EPA, precursor to PGE3) on IL-2 production by PHA-stimulated human PBMC. DGLA and AA inhibited IL-2 production in a dose-dependent manner: half-maximal inhibition was obtained by using the fatty acids at the dose of 10 micrograms/ml without significant effects on cell viability. EPA inhibited IL-2 production by PBMC of only some donors. Incubation of cells in the presence of oleic, stearic, and palmitic acids, which are not PG precursors, did not affect mitogen-induced IL-2 production. A progressive increase in incorporation of DGLA into cellular lipids was observed over a 48-h incubation period. IL-2 production was reduced also when PBMC were pretreated overnight with DGLA or AA and washed before exposure to PHA. Whereas addition of the cyclo-oxygenase inhibitor, indomethacin, at the time of mitogenic stimulation led to increased IL-2 production and prevented mitogen- and fatty acid-induced increases in PGE release, it had no significant effect on the capacity of the fatty acids to suppress IL-2 production. Time course experiments showed that DGLA and AA inhibited IL-2 production even at times of minimal or no PGE release by the treated cultures. Moreover, DGLA and AA inhibited IL-2 production by the human leukemia T cell line Jurkat which, when appropriately induced, is able to release high levels of IL-2 in the absence of accessory cells and measurable PGE production. Taken together, these data indicate that essential fatty acids inhibit IL-2 production directly without conversion into their cyclo-oxygenase pathway products, and suggest that human lymphocyte function may be altered profoundly by small changes in their fatty acid profile. PMID- 2545789 TI - Detection of herpes simplex virus DNA in cutaneous lesions of erythema multiforme. AB - The association between erythema multiforme (EM) and herpes simplex virus (HSV) infection has long been appreciated, although the exact role which HSV may play in the pathogenesis of this herpes-associated EM (HAEM), is unknown. Previous studies have suggested, but not definitively demonstrated, the presence of HSV in lesions of HAEM. The presence of HSV would support the hypothesis that an immune mediated response directed against HSV-specific antigens in the skin is central to lesion development in HAEM. The purpose of this study was to examine lesions of EM for the presence of HSV DNA by using the polymerase chain reaction (PCR). In addition, in situ hybridization using an HSV-specific RNA probe was performed to further localize the HSV nucleic acids within the skin. DNA was extracted from formalin-fixed, paraffin-embedded specimens of cutaneous lesions of HAEM and also from EM for which no precipitating factor could be documented, otherwise known as idiopathic EM (IPEM). DNA from lesions of bullous pemphigoid served as a negative control. Using PCR to specifically amplify HSV sequences which might be present, and then performing Southern analysis, we demonstrated HSV DNA in 9/13 HAEM and 6/9 IPEM biopsies. No HSV was detected in six lesions of bullous pemphigoid. In situ hybridization of three cutaneous HAEM lesions using an 35S-labeled HSV specific RNA probe localized the HSV nucleic acids predominantly to the epidermis. Three biopsies of chronic dermatitis, used as negative controls, did not demonstrate this specific hybridization. These findings confirm the presence of HSV in lesions of HAEM and are consistent with the concept of an HSV-specific immune-mediated pathogenesis for this disease. In addition, most cases of IPEM appear to be herpes associated despite the absence of clinically apparent HSV infection. PMID- 2545790 TI - Cutaneous graft-versus-host reaction lacks evidence of cutaneous cytomegalovirus by the immunoperoxidase technique. AB - Sixty skin biopsy specimens from 21 bone-marrow transplant patients were evaluated for the presence of cytomegalovirus (CMV) using two monoclonal antibodies to early and late antigens. Each patient had at least one biopsy showing an acute graft-versus-host reaction (GVHR), grade 2, and one positive culture for CMV from blood, bone marrow or urine. In no case could CMV antigens be identified in biopsies showing an acute or chronic cutaneous GVHR or in any other of the skin biopsies obtained from these patients. While CMV may play a role in immunologic events culminating in graft-versus-host disease (GVHD), this immunoperoxidase study did not reveal evidence of viral antigens in tissue displaying features of cutaneous GVHR. PMID- 2545792 TI - Comparative susceptibility of respiratory viruses to recombinant interferons alpha 2b and -beta. AB - Intranasal recombinant interferon-alpha 2b (rIFN-alpha 2b) protects against natural colds due to rhinoviruses, but apparently not against those caused by viruses. Because rIFN-beta serine17 (rIFN-beta ser) appears less active than rIFN alpha 2b in preventing natural rhinovirus colds, we compared the two IFNs in two in vitro assays against selected respiratory viruses. In a yield reduction assay, both IFNs had comparable activity against rhinovirus types 39 and 1A and coronavirus 229E, which were inhibited by 90% or more at IFN concentrations of 10(-11) to 10(-10) gram of protein/ml (approximately 2-20 IU/ml). Similar activities were observed with rIFN-beta ser against rhinoviruses isolated from clinical specimens. At concentrations of 10(-9) gram protein/ml, both IFNs inhibited the growth of influenza A and parainfluenza viruses, but not of adenovirus or respiratory syncytial virus in the cell culture systems tested. Thus, the different clinical protection conferred by rIFN-alpha 2b and rIFN-beta ser in studies of natural rhinovirus colds are not accounted for by differences in their in vitro activity against these viruses, and other explanations must be found. PMID- 2545791 TI - The in vivo antiviral effect of CL246,738 is mediated by the independent induction of interferon-alpha and interferon-beta. AB - An interferon (IFN) inducer and immunomodulator, CL246,738 [3,6-bis(2 piperidinoethoxy)acridine trihydrochloride], protected mice from lethal infection with Semliki Forest (SFV) and Banzi (BZV) viruses. A single oral dose of CL246,738 (5-150 mg/kg) administered 24 h before intraperitoneal challenge with SFV or BZV fully protected mice from lethal infection. Dose-dependent levels of circulating IFN peaked at 24 h in the serum and peritoneal fluid of CL246,738 treated mice. The circulating IFN of CL246,738-treated mice consisted of IFN alpha and was produced by spleen cells. Peritoneal exudate cells (PEC) obtained from CL246,738-treated mice produced IFN-beta. Treatment in vivo with anti-IFN alpha/beta and anti-IFN-beta reversed the protective effect of CL246,738 against lethal SFV encephalitis. PMID- 2545793 TI - Value of ER-D5 immunocytochemical determination in routine tissue sections of human breast cancer. AB - ER-D5 is a recently identified protein related to estrogen receptors (ER). Generally ER measurement requires fresh frozen tissue and for ER-D5 assay ethanol (E) fixation of the specimen is recommended. We evaluated the possibility of immunocytochemical detection of ER-D5 in routine formalin-fixed (F) sections in 51 breast cancers comparing the results with those obtained in the same specimens using E as fixative. The results of ER-D5 assay were expressed by the staining index (SI) taking values greater than or equal to 5 as positive. In all tumors ER was also assayed by a biochemical method (DCCA). The sensitivity of ER-D5 detection in F was only 33.3%, while the specificity was 94.4%. A lower cut-off value of SI for F sections (greater than or equal to 2) increased the sensitivity to 66.6%, leaving the specificity unchanged. A strong correlation was found between the SI of ER-D5 in E and F (p less than 0.001). The SI of ER-D5 in F sections was also well correlated with ER concentrations (p less than 0.001). These results suggest that immunocytochemical determination of ER-D5 in routine sections may be useful in retrospective studies of hormone dependence in breast cancer. PMID- 2545794 TI - Carcinoembryonic antigen levels in germ cell tumours. AB - Elevated serum carcinoembryonic antigen (CEA) prior to specific treatment was noted in 3% (7/258) of assessable patients with testicular, extragonadal or ovarian germ cell tumours (GCT). In addition, persistently raised CEA was documented in 7% (26/385) of patients during or after cisplatin-based chemotherapy for metastatic GCT. Raised CEA did not appear associated with adverse prognosis. Among patients undergoing resection of residual tumour masses post-chemotherapy, 8 of 36 with mature differentiated teratoma excised had raised CEA compared with only one of 39 patients where no mature teratoma was found. However, CEA levels remained elevated in 6 of the 8 cases despite apparent complete resection of mature teratoma. Elevated CEA in treated GCT patients may be caused by hepatotoxicity from chemotherapy, intercurrent diseases, or other unknown factors. History of cisplatin-based chemotherapy may be a confounding factor in interpreting raised CEA levels. CEA measurements do not help in the management of patients with germ cell tumours. PMID- 2545796 TI - Pathogenic effect of osteopetrosis virus strain MAV-2(0) on guinea fowl pancreas. AB - Osteopetrosis virus strain MAV-2(0) in guinea fowl pancreas induces early unspecific changes in separate acini and the development of neoplasia among apparently unaffected acinar tissue. The tumors were classified as serous and mucinous adenomas and adenocarcinomas, as well as poorly differentiated carcinomas. Foci of phenotypically altered acinar cells were observed in single experimental birds. PMID- 2545797 TI - [Japanese encephalitis virus infection in cattle: a serological survey in Saitama and Kagoshima Prefectures]. AB - Serum samples were collected from a total of 1,306 and 536 cattle raised in Saitama and Kagoshima Prefectures, respectively, during a period from 1982 to 1984. Their antibody titers against Japanese encephalitis virus (JEV) were determined by the hemagglutination-inhibition test. Antibody prevalences were 65.5% (856/1306) and 68.8% (369/536) in Saitama and Kagoshima Prefectures, respectively. In both prefectures, there was a rapid increase in positive rate and mean titer in summer. Afterwards, there was a gradual decrease in both rate and titer in Saitama Prefecture in winter. In Kagoshima Prefecture, however, the rate and titer did not so markedly decrease in winter, but remained high in December. The rate of positive reactors to JEV in Kagoshima Prefecture increased gradually from 29.4% (1 year old) to 74.5% (4 years old) with advance in age. This rate in each age group remained almost at the same level ranging from 64.0% to 82.8% in Saitama Prefecture. PMID- 2545795 TI - Opioid receptors in human neoplasms: growth hormone and prolactin response to morphine, met-enkephalin and buprenorphine in cancer patients. PMID- 2545798 TI - [Localization of a variant of herpes simplex virus type 1 in the western Japan]. AB - We previously reported that a variant of herpes simplex virus type 1, which we found by Bg1 II digestion of many Japanese HSV-1 isolates' DNA's, was isolated frequently in two regions of Western Japan, i.e. Yamaguchi and Fukuoka. In this paper, we analyzed 280 more isolates from eleven regions in Japan. In four western regions of Japan, i.e. Osaka, Shimane, Yamaguchi, and Fukuoka, the isolation frequency of the variant was very high, and its proportion among all the isolates from those regions was 30 to 50%. On the other hand, in four eastern regions of Japan, i.e. Akita, Iwate, Miyagi, and Yamanashi, the proportion was 0 to 11%. In two central regions, i.e. Aichi and Kyoto, the proportions were 16 and 22%, respectively. Thus, the variant seems to be localized in the western regions of Japan. PMID- 2545799 TI - [SEM study on Chlamydia trachomatis in cultured McCoy cell--with special reference to surface projections and intra-inclusion network]. AB - A study on the growth cycle of C. trachomatis of genital origin in McCoy cells has produced the following results: 1) Two kinds of surface projections, spikelike and hemispheric, were observed. The spikelike projections were recognized 50 hours after infection in larger C. trachomatis of 1.0 micron in diameter. From the surface these C. trachomatis measured 21-52 nm in height with diameters of 25-35 nm. Hemispheric projections were observed 72 hours after infection in smaller C. trachomatis of under 0.3 micron in diameter. Their average diameter was 34.0 nm. These projections were hexagonally arranged with the distances between the center of adjacent projections at 40-45 nm. 2) Intra inclusion networks were observed. These were recognized in inclusions 72 hours after infection. Their structure was three dimensional with lineal materials extending from one C. trachomatis to another. The networks were composed of granular particles averaging 53.9 nm in diameter. These granular particles were not noticed after amylase treatment and lineal material of 33.5 nm in width was noticed instead. This lineal material may be regarded as the axis of the network. Structurally, the network is an axis 33.5 nm wide with glycogen granules attached to the perimeter. The network structures were classified into long, short, and inclusion types. PMID- 2545801 TI - Nerve damage in leprosy. PMID- 2545800 TI - [Molecular genetics of Fabry disease]. PMID- 2545802 TI - [Insulinoma simultaneously secreting calcitonin and calcitonin gene-related peptide. A case report]. PMID- 2545803 TI - [A case of severe hepatitis A with positive IgM antibody to hepatitis A virus of long duration, monoclonal gammopathy (IgG.lambda) and hemolytic anemia]. PMID- 2545804 TI - Syndactyly: a review of the factors which influence surgical treatment. AB - A review was undertaken of 100 patients with 218 surgically-treated syndactylies operated upon over a ten year period. The pre-operative factors which influenced the result of surgical treatment were the complexity of the syndactyly and the presence of other congenital abnormalities in the hand. The operative factor which most influenced outcome was the type of skin-graft used. 42 patients required at least one secondary operation to obtain an acceptable result, comprising 22% who developed web creep and 26% a significant flexion contracture; five patients developed both a contracture and web creep. In contrast, the type of flap used to reconstruct the web floor had little influence on the result. PMID- 2545805 TI - Full thickness grafts taken from the plantar instep for syndactyly release. AB - Thirteen digital clefts (eight patients) were reviewed in which full-thickness plantar instep skin was used in syndactyly release. The results are presented, emphasising the good colour match of plantar instep skin with digital skin. There was a surprising lack of morbidity in the thirteen donor sites. PMID- 2545807 TI - Mitochondrial biogenesis during the activation of lymphocytes by mitogens: the immunosuppressive action of tetracyclines. AB - The role of mitochondrial biogenesis and function during mitogenic stimulation of rat thymocytes was investigated. The results show that mitochondrial biogenesis is required to provide the ATP for the energy-requiring processes occurring during blastogenesis. Impairment of mitochondrial biogenesis by inhibition of mitochondrial protein synthesis inhibits blast transformation. Since the tetracyclines impair mitochondrial protein synthesis, the results offer an explanation for the well-known immunosuppressive effects of these antibiotics. PMID- 2545806 TI - Do aggressive subclones within primary colorectal cancer give rise to liver metastases? AB - Primary colorectal cancers and hepatic metastases from 24 patients have been compared according to type, grade, DNA content as measured by flow cytometry, and expression of two differentiation antigens and carcinoembryonic antigen (CEA). The primary and secondary tumours were histologically indistinguishable in 23 out of 24 cases. DNA content differed between primary and metastasis in only six cases. Immunohistochemical profiles were usually the same in the primary and metastasis. In particular, heterogeneous expression by the primary was mirrored in the metastasis. In 17 out of 24 (70.1%) patients the primary and metastasis were indistinguishable by all six parameters. In one of these, the liver metastasis had been removed 14 years 10 months after the primary tumour. It is concluded that DNA content as measured by flow cytometry may remain stable for long periods and that the differences shown in this study may simply be due to inadequate sampling of the primary tumour. Phenotypic heterogeneity may be epigenetic and not indicative of genetic instability and cannot be used to track the progression of subclones. PMID- 2545808 TI - Arachidonic acid metabolism in guinea pig eosinophils: synthesis of thromboxane B2 and leukotriene B4 in response to soluble or particulate activators. AB - The arachidonic acid metabolism of guinea pig eosinophils isolated from either peritoneal cavity or bronchoalveolar lavages was studied by reverse-phase high performance liquid chromatography. The purified eosinophils (95-100%) from either source released thromboxane B2 (TxB2), luekotriene B4 (LTB4) and 5-hydroxy eicosatetraenoic acid (5-HETE) following calcium ionophore A23187 stimulation. Quantification by radioimmunoassay indicated that maximal mediator output from the stimulated peritoneal cells was reached at 3 min after stimulation. The increase in production of TxB2 and LTB4 was correlated to increasing calcium ionophore A23187 concentration up to 1.0 micrograms/ml. In addition to calcium ionophore, the guinea pig peritoneal cells were also activated by f-met-leu-phe, phorbol 12-myristate, 13-acetate (PMA), and to lesser extent platelet-activating factor (PAF) to produce TxB2. LTB4 synthesis was stimulated by calcium ionophore, by f-met-leu-phe, as well as by unopsonized glucan, a particulate phagocytotic stimulus. The guinea pig eosinophils do not synthesize sulfidopeptide leukotrienes because of the absence of the specific LTA4 glutathione S transferase. These results suggest that the guinea pig eosinophils differ from the human circulating eosinophils in the synthetic capacity of lipid mediators derived from arachidonic acid metabolism. This difference may be important in the understanding of the role of the eosinophils in inflammatory reactions such as that which occurs in the bronchial tissues of asthmatics. PMID- 2545809 TI - Effect of granulocyte-macrophage colony-stimulating factor on superoxide production in cytoplasts and intact human neutrophils: role of protein kinase and G-proteins. AB - Granulocyte-macrophage colony-stimulating factor, GM-CSF, potentiates superoxide generation produced by human neutrophils stimulated with fMet-Leu-Phe and platelet-activating factor, PAF, but not by phorbol 12-myristate 13-acetate (PMA) or opsonized zymosan. The potentiation is greatest in fMet-Leu-Phe-stimulated cells. This indicates that the actions of only certain receptors are potentiated by GM-CSF. Incubation of the cells with the protein kinase inhibitor H-7 or with the protein synthesis inhibitor cyclohexamide before the addition of GM-CSF does not affect the observed potentiation. The rationales behind these studies are to examine the roles of protein kinase C and protein synthesis in the action of GM CSF. The data suggest that neither protein kinase C nor protein synthesis is necessary for GM-CSF action. On the other hand, no potentiation can be seen in the presence of cytochalasin B. Unlike intact cells, GM-CSF does not enhance superoxide production by cytoplasts stimulated with fMet-Leu-Phe. The rationale behind the use of cytoplasts is to examine the role of granules and/or nucleus in GM-CSF action, and the data indicate that one or more of these two components is necessary for the priming effect of GM-CSF. The amount of actin associated with the cytoskeleton under control of fMet-Leu-Phe-stimulated condition is the same in normal and GM-CSF-treated human neutrophils. Botulinum D toxin ADP-ribosylates a protein with a molecular weight of 22 kDa. This ribosylation is reduced in homogenates obtained from cells pretreated with botulinum D toxin or GM-CSF. Botulinum D toxin does not affect the basal or the fMet-Leu-Phe-induced rise in the intracellular concentration of free calcium in human neutrophils. GM-CSF also increases the rise in intracellular concentration of free calcium in human neutrophils stimulated with PAF or fMet-Leu-Phe. The increases are inhibited by pertussis toxin. Several important conclusion can be drawn from these data. 1) GM CSF potentiates the rise in Ca2+i produced by PAF and fMet-Leu-Phe, and these potentiations are inhibited in pertussis-toxin-treated cells. 2) GM-CSF does not prime cytoplasts to stimulation by fMet-Leu-Phe. This suggests that the granules and/or nucleus are necessary for the priming action. 3) The priming by GM-CSF is not mediated by the H-7-sensitive protein kinase C, botulinum D-sensitive G protein, or protein synthesis. PMID- 2545810 TI - [Expressivity of familial forms of Fraser syndrome]. AB - Two new cases of the Fraser syndrome are presented. The literature review indicates that the clinical expression inside families is very constant with regard to cryptophthalmos, syndactyly or internal malformations. Particularly the fatal issue of severe forms is regularly constant in each of the families recently reported with a detailed autopsy. These conclusion would change the genetic counseling. A lethal form of the disease would recur in the same manner and could be detected by exhibiting the genito-urinary malformations at ultrasonography or even the syndactyly at foetoscopy. PMID- 2545811 TI - [Prenatal diagnosis of X-linked adrenal hypoplasia associated with glycerol kinase deficiency]. AB - We report a case of X-linked adrenal hypoplasia associated with glycerol kinase deficiency in a boy. Cytogenetic studies and X-linked probes did not demonstrate deletion at Xp21. These probes are not informative enough to be used in prenatal diagnosis. This diagnosis was achieved by glycerol concentration assay in amniotic fluid and by maternal plasma estriol assay. PMID- 2545812 TI - Subsensitivity of serotonin and substance P receptors involved in nociception after repeated administration of a serotonin receptor agonist. AB - The antinociceptive effects of subcutaneous 5-methoxy-N,N-dimethyltryptamine (5 MeODMT) and 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) and the responses to intrathecal (i.th.) serotonin (5-HT) and substance P (SP) were examined in mice after repeated administration of 5-MeODMT (3 mg/kg every 30 min for 4 hours). Ninety min after the last injection of 5-MeODMT the basal tail-flick and hot-plate response latencies were unaltered, but the antinociceptive effects of 5 MeODMT (3 mg/kg) in the tail-flick and hot-plate tests and the antinociceptive effect of 8-OH-DPAT (0.5 mg/kg) in the hot-plate test were markedly reduced. The behavioral responses to i.th. 5-HT (4.0 micrograms) and SP (2.5, 5, and 10 ng) which include vigorous biting, licking and scratching of the caudal part of the body, were attenuated 90-120 min after withdrawal of 5-MeODMT treatment. It is suggested that repeated administration of 5-MeODMT downregulates the function of the 5-HT receptors mediating the antinociceptive effects of 5-MeODMT and 8-OH DPAT. The rapid desensitization to the behavioral responses both to 5-HT and SP by 5-MeODMT pretreatment may reflect a functional interaction between 5-HT and SP in the spinal modulation of nociception. PMID- 2545813 TI - Dramatic synergism between MK-801 and clonidine with respect to locomotor stimulatory effect in monoamine-depleted mice. AB - The present study corroborates previous findings showing that the selective, non competitive N-methyl-D-aspartate (NMDA) antagonist MK-801 [(+)-5-methyl-10,11 dihydroxy-5H-dibenzo(a,d)-cyclohepten-5,10-imi ne] produces a dose-dependent increase in locomotion in mice pretreated with a combination of the monoamine depleter reserpine and the catecholamine synthesis inhibitor alpha-methyl-para tyrosine. Moreover, the present investigation demonstrates a synergistic interaction between MK-801 and the alpha-adrenergic agonist clonidine in monoamine-depleted mice: MK-801 in a dose of 1 mg/kg and clonidine in a dose of 2 mg/kg hardly affected locomotion when given separately, but when the two drugs were combined a dramatic enhancement of motor activity was observed. This effect was effectively antagonized by the alpha 2-adrenergic blockers idazoxan and yohimbine, as well as by the "atypical" neuroleptic clozapine. Likewise, a clear cut synergism was observed when a low dose of the dopamine receptor agonist apomorphine (0.1 mg/kg), which did not per se affect motor activity, was combined with MK-801 (1.5 mg/kg); however, the synergism between apomorphine and MK-801 was less dramatic than that observed between MK-801 and clonidine. The results may have important neuropsychiatric implications related to, e.g. the treatment of Parkinson's disease and the pathogenesis of schizophrenia. PMID- 2545814 TI - Laser microbeam axotomy and conduction block show that electrical transmission at a central synapse is distributed at multiple contacts. AB - Touch (T) sensory neurons in the leech innervate defined regions of skin and synapse on other neurons, including other T cells, within the ganglionic neuropil. The cells' receptive fields in the periphery are comprised of a central region, innervated by thick axons, and adjoining regions (minor fields) innervated by thinner axons. Secondary branches, known to be sites of synapses, emerge from the thinner and thicker axons. Pairs of T cells appear to make up to 200 separate contacts distributed within the neuropil. When the T cell is hyperpolarized, as occurs during natural stimulation of the cell, action potentials generated in the minor field and travelling into the ganglion along the thin axons may fail to conduct at central branch points. Evidence is presented, using axon conduction block and laser axotomy of cells filled with 6 carboxy-fluorescein, that synapses between separate groups of branches can function independently. Thus, selective activation of branches of the thin anterior axon produced a synaptic potential 36 +/- 6% of control amplitude, which was consistent with counts of 39 +/- 6% of contacts made by these branches. Laser axotomy of postsynaptic neurons showed that the anterior contacts indeed made the principal or only contacts activated during anterior conduction block. The results show that conduction block can modulate transmission within the ganglion, and it operates by silencing particular contacts between cells. PMID- 2545815 TI - Subtype-selective immunoprecipitation of the beta 2-adrenergic receptor. AB - Most antibodies known to interact with beta-adrenergic receptors do not exhibit subtype selectivity, nor do they provide quantitative immunoprecipitation. A monoclonal antibody, G27.1 raised against a synthetic peptide corresponding to the C-terminus of the beta 2-adrenergic receptor of hamster, is selective for the beta 2 subtype. G27.1 provides nearly quantitative immunoprecipitation of the beta 2-adrenergic receptor from hamster lung that has been photoaffinity-labeled and solubilized with sodium dodecyl sulfate. Immunoprecipitation is completely blocked by nanomolar concentrations of the immunizing peptide. This antibody interacts with beta 2-adrenergic receptors from three rodent species, but not with those from humans. When C6 glioma cells, which contain both beta 1- and beta 2-adrenergic receptors, are photoaffinity-labeled in the absence or presence of subtype-selective antagonists, subtype-selective photoaffinity-labeling results. G27.1 can immunoprecipitate beta 2-, but not beta 1-, adrenergic receptors from these cells. Similar results were obtained following subtype-selective photoaffinity-labeling of membranes from rat cerebellum and cerebral cortex. The beta-adrenergic receptors from C6 glioma cells and rat cerebral cortex exist as a mixture of two molecular weight species. These species differ in glycosylation, as shown by endoglycosidase F digestion of crude and immunoprecipitated receptors. PMID- 2545816 TI - Characterization of a [3H]glycine recognition site as a modulatory site of the N methyl-D-aspartate receptor complex. AB - A [3H]glycine recognition site in rat brain synaptic plasma membranes (SPM) has been identified, having characteristics expected of a modulatory component of the N-methyl-D-aspartate receptor complex. Incubation of SPM with [3H]glycine for 10 min at 2 degrees C results in saturable, reversible binding with a KD of 0.234 microM and a Bmax of 9.18 pmol/mg. A pharmacological analysis of this binding site indicates that D-serine (Ki = 0.27 microM), D-alanine (Ki = 1.02 microM), and D-cycloserine (Ki = 2.33 microM) are potent inhibitors of binding, whereas the corresponding L isomers have significantly less activity (Ki = 25.4 microM, 15.9 microM, and greater than 100 microM, respectively). Inactive at concentrations of up to 100 microM were strychnine, L-valine, N,N dimethylglycine, aminomethylphosphonate, and aminomethylsulfonate. The active compounds were analyzed further for their ability to stimulate [3H]1-[1-(2 thienyl)cyclohexyl]piperidine [( 3H]TCP) binding to Triton X-100-washed SPM. Results indicate that the affinity of the compounds for the [3H]glycine recognition site correlates with the ability of these analogues to stimulate [3H]TCP binding. PMID- 2545817 TI - Kinetic analysis of A23187-mediated polyphosphoinositide breakdown in rat cortical synaptosomes suggests that inositol bisphosphate does not arise primarily by degradation of inositol trisphosphate. AB - The kinetics of polyphosphoinositide breakdown and inositol phosphate formation have been studied in rat cortical synaptosomes labelled in vitro with myo-[2 3H]inositol. Intrasynaptosomal Ca2+ concentrations have been varied by the use of Ca-EGTA buffers or by adding the ionophore A23187 in the presence and absence of 1 mM Ca2+. The former studies have revealed that, at very low (20 nM) intrasynaptosomal free Ca2+ levels, inositol bisphosphate, but not inositol monophosphate levels are reduced. Addition of A23187 in the absence of added Ca2+ gives rise to greatly enhanced inositol bisphosphate accumulation, which is further enhanced if 1 mM Ca2+ is present in the extrasynaptosomal medium. At all time points examined (down to 2 s after adding ionophore), the ratio of inositol trisphosphate/inositol bisphosphate accumulation does not exceed 0.2, and calculations based on inositol bis- and trisphosphate breakdown rates in synaptosomal lysates suggest that only a minority of the inositol bisphosphate arises from degradation of inositol trisphosphate. Addition of ionophore in the presence (but not in the absence) of 1 mM Ca2+ leads to rapid breakdown of phosphatidylinositol 4,5-bisphosphate (PtdInsP2) and ATP and slower breakdown of phosphatidylinositol 4-phosphate (PtdInsP). The rates of loss of PtdinsP2 and ATP are very highly correlated, suggesting that polyphosphoinositide resynthesis may be limited by ATP availability at high Ca2+ levels. Analysis of 32P-labelled synaptosomes also reveals that A23187 produces Ca2+-dependent losses of PtdInsP2, PtdInsP, ATP, and GTP radioactivity and a marked increase in the radioactivity of a compound distinct from nucleotides or any of the lipid breakdown products tested.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545818 TI - Glycine binding to rat cortex and spinal cord: binding characteristics and pharmacology reveal distinct populations of sites. AB - Glycine is the principal inhibitory neurotransmitter in posterior regions of the brain. In addition, glycine serves as an allosteric regulator of excitatory neurotransmission mediated by the N-methyl-D-aspartate (NMDA) acidic amino acid receptor subtype. The studies presented here characterize [3H]glycine binding to washed membranes prepared from rat spinal cord and cortex, areas enriched in glycine inhibitory and NMDA receptors, respectively, in an attempt to define the glycine recognition sites on the two classes of receptors. Specific binding for [3H]glycine was seen in both cortex and spinal cord. Saturation analyses in cortex were best fitted by a two-site model with respective equilibrium dissociation constants (KD values) of 0.24 and 5.6 microM and respective maximal binding constants (Bmax values) of 3.4 and 26.7 pmol/mg of protein. Similar analyses in spinal cord were best fitted by a one-site model with a KD of 5.8 microM and Bmax of 20.2 pmol/mg of protein. Na+ had no effect on [3H]glycine binding to cortical membranes but increased the binding to spinal cord membranes by greater than 15-fold. This Na+-dependent binding may reflect glycine binding to the recognition site of the high-affinity, Na+-dependent glycine uptake system. Several short-chain, neutral amino acids displaced [3H]glycine binding from both cortical and spinal cord membranes. The most potent displacers of [3H]glycine binding to cortical membranes were D-serine and D-alanine, followed by the L-isomers of serine and alanine and beta-alanine. In contrast, D-serine and D-alanine were similar in potency to L-serine in spinal cord membranes. Compounds active at receptors for the acidic amino acids had disparate effects on the binding of [3H]glycine. At 10 microM, NMDA resulted in a 25% increase, whereas D- and L-2-amino-5-phosphonovaleric acid at 100 microM resulted in a 30% decrease, in [3H]glycine binding to cortical membranes. Kynurenic acid was the most potent of the acidic amino acid-related compounds at displacing [3H]glycine binding. In cortical membranes, kynurenic acid displacement was resolved into a high- and a low-affinity component; the high-affinity component displaced the high-affinity component of [3H]glycine binding.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2545819 TI - Modulation of histamine release and synthesis in the brain mediated by alpha 2 adrenoceptors. AB - The adrenergic regulation of histamine release was studied in rat brain slices labeled with L-[3H]histidine. Noradrenaline in increasing concentrations progressively inhibited K+-evoked [3H]histamine release from cortical slices, whereas phenylephrine and isoprenaline were ineffective. Yohimbine, a preferential alpha 2-adrenoceptor antagonist, reversed the noradrenaline effect in an apparently competitive manner and with a mean Ki value of 30 nM. Phentolamine reversed the noradrenaline effect with a similar potency, whereas propranolol was ineffective. The imidazolines clonidine and oxymetazoline acted as partial agonists, oxymetazoline even behaving as an apparent antagonist. In vivo clonidine also inhibited [3H]histamine formation in cerebral cortex, an effect reversed by the administration of yohimbine. However, yohimbine failed to increase significantly [3H]histamine release in vitro and [3H]histamine formation in vivo, suggesting that adrenergic receptors are not activated by endogenous noradrenaline released under basal conditions. It is concluded that adrenergic alpha 2-adrenoceptors presumably located on histaminergic axons control release and synthesis of histamine in the brain. PMID- 2545820 TI - Effects of lithium on inositol phospholipid hydrolysis and inhibition of dopamine D1 receptor-mediated cyclic AMP formation by carbachol in rat brain slices. AB - The in vitro and ex vivo effects of lithium on muscarinic cholinergic inositol phospholipid hydrolysis and muscarinic cholinergic inhibition of dopamine D1 receptor-stimulated cyclic AMP formation were examined in rat brain slices. Following chronic lithium feeding, carbachol-stimulated inositol phosphate accumulation was reduced ex vivo in slices of cerebral cortex but not in striatal slices. Lithium (1 mM) in vitro had no direct effect on dopamine D1-receptor stimulated cyclic AMP formation, but enhanced the inhibitory effect of carbachol on the D1 response, in striatal slices, and this was not significantly altered by prior lithium feeding. Lithium therefore has effects on two discrete muscarinic responses in rat brain which are apparently maintained after chronic exposure to the ion and might be relevant to its antimanic actions. PMID- 2545821 TI - No role for phospholipase A2 and protein kinase C in the potentiation by alpha adrenoceptors of beta-adrenoceptor-mediated cyclic AMP formation in rat brain. AB - This study was undertaken to examine the role of phospholipase A2 and protein kinase C in the potentiation of beta-adrenoceptor-mediated cyclic AMP formation by alpha-adrenoceptors in rat cerebral cortical slices. Inhibition of arachidonic acid metabolism by a range of cyclooxygenase and lipoxygenase inhibitors had no effect on the potentiation of isoprenaline-stimulated cyclic AMP. Conversely, stimulation of leukotriene formation had no effect on the response to isoprenaline. The phospholipase A2 activator, melittin, stimulated cyclic AMP and potentiated the effect of isoprenaline, but these responses were not influenced by cyclooxygenase or lipoxygenase inhibitors. Indomethacin was also ineffective against the potentiation of vasoactive intestinal peptide-stimulated cyclic AMP by noradrenaline. Phorbol ester potentiated the cyclic AMP response to isoprenaline, and this potentiation was antagonized by three different putative protein kinase C inhibitors. However, the same inhibitors did not affect the alpha-adrenoceptor-stimulated enhancement of the response to isoprenaline. We have found no evidence, therefore, to support the suggestion that arachidonic acid and its metabolites and/or protein kinase C mediate the alpha-adrenoceptor modulation of beta-adrenoceptor function. PMID- 2545822 TI - In vivo determination of 5-hydroxytryptamine receptor-stimulated phosphoinositide turnover in rat brain. AB - Phosphoinositide turnover stimulated by 5-hydroxytryptamine (5-HT) receptors in the intact rat brain was studied using an in vivo method. Phosphoinositides in the rat brain were prelabeled with [3H]inositol injected into the lateral cerebral ventricles. The rats were killed by microwave irradiation after 48 h and the contents in the frontal cortex of 3H-inositol phosphates, [3H]inositol-1 monophosphate [( 3H]IP1), [3H]inositol-1,4-bisphosphate [( 3H]IP2), and a mixture of [3H]inositol-1,4,5-trisphosphate and [3H]inositol-1,3,4-trisphosphate [( 3H]IP3) were assayed by HPLC. Lithium treatment (10 mEq/kg, i.p., 2 h before) increased the content of [3H]IP1 and [3H]IP2. 5-Methoxy-N,N-dimethyltryptamine (5 MeODMT) and quipazine, 5-HT agonists, significantly increased the amount of 3H inositol phosphates under lithium pretreatment. The response to 5-MeODMT was inhibited by ritanserin, a 5-HT2 antagonist, but not by (-)-propranolol, a 5-HT1 antagonist. These results suggest that phosphoinositide turnover in the rat frontal cortex in vivo is stimulated by 5-HT2 receptor activation. It is considered that this method will be useful for measurement of 5-HT2 receptor stimulated phosphoinositide turnover in vivo to examine the in vivo effects of various psychotropic drugs such as antidepressants. PMID- 2545823 TI - Chronically administered lithium alters neither myo-inositol monophosphatase activity nor phosphoinositide levels in rat brain. AB - Rats were exposed to either 29 consecutive days of LiCl injections or 27 and 39 days of dietary Li2CO3, followed by injected LiCl at the end of the diet to insure a constant level of exposure to the drug. At the end of the period of chronic exposure to lithium, the rats were sacrificed and brain myo-inositol-1 phosphate phosphohydrolase (myo-inositol monophosphatase) activity was measured. In none of the experiments was there any difference in the lithium-sensitive activity toward myo-inositol monophosphatase when comparing the control and chronic groups. These brains and those from another group of rats that had been given Li2CO3 in their diet for 41 days, followed by 7 additional days of LiCl injections, were also examined for changes in the levels of the phosphoinositides. No reproducible differences in the absolute tissue levels of those lipids were found when control and chronic lithium groups were compared. These results are contrary to published reports which suggest that myo-inositol monophosphatase activity increases and that the phosphatidylinositol level decreases in rat brain as a result of chronic administration of lithium. PMID- 2545824 TI - Alpha 2-adrenergic and muscarinic cholinergic receptors have opposing actions on cyclic AMP levels in SK-N-SH human neuroblastoma cells. AB - Forskolin and vasoactive intestinal polypeptide (VIP) were shown to increase cyclic AMP accumulation in a human neuroblastoma cell line, SK-N-SH cells. The alpha 2-adrenergic agonist UK 14304 decreased forskolin-stimulated cyclic AMP levels by 40 +/- 2%, with an EC50 of 83 +/- 20 nM. This response was blocked by pretreatment with pertussis toxin (PT) (EC50 = 1 ng/ml) or by the alpha 2 antagonists yohimbine, idazoxan, and phentolamine. Antagonist IC50 values were 0.3 +/- 0.1, 2.2 +/- 0.3, and 1.4 +/- 0.1 microM, respectively. This finding suggests the presence of normal inhibitory coupling of SK-N-SH cell alpha 2 adrenergic receptors to adenylate cyclase via the inhibitory GTP-binding protein species, Gi. Muscarinic receptors in many target cell types are coupled to inhibition of adenylate cyclase. However, in SK-N-SH cells, muscarinic agonists synergistically increased (67-95%) the level of cyclic AMP accumulation elicited by forskolin or VIP. EC50 values for carbamylcholine (CCh) and oxotremorine facilitation of the forskolin response were 1.2 +/- 0.2 and 0.3 +/- 0.1 microM, respectively. Pharmacological studies using the muscarinic receptor subtype preferring antagonists 4-diphenylacetoxy-N-methylpiperidine, pirenzepine, and AF DX 116 indicated mediation of this response by the M3 subtype. IC50 values were 14 +/- 1, 16,857 +/- 757, and 148,043 +/- 16,209 nM, respectively. CCh-elicited responses were unaffected by PT pretreatment. Muscarinic agonist binding affinity was indirectly measured by the ability of CCh to compete for [3H]quinuclidinyl benzilate binding sites on SK-N-SH cell membranes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545825 TI - Age-related loss of t-[35S]butylbicyclophosphorothionate binding to the gamma aminobutyric acidA receptor-coupled chloride ionophore in rat cerebral cortex. AB - Muscimol and t-butylbicyclophosphorothionate (TBPS) are known to label two distinct sites within the gamma-aminobutyric acidA (GABAA) receptor complex, i.e., the GABA recognition site and the chloride ionophore, respectively. Age dependent changes in the specific binding of [3H]muscimol and [35S]TBPS were compared in membranes prepared from the cerebral cortex of rats, 2-800 days old. Perinatal (day 2) binding of muscimol and TBPS represented 8 and 20% of the respective values for adults (day 180). After the first week, muscimol binding increased more rapidly than TBPS binding. Levels near those of adults were reached at day 20 and remained practically unchanged in adulthood (day 180). In aged (780-day-old) rats, the binding of TBPS was significantly reduced, whereas muscimol binding did not change compared with adult values. This decrease of TBPS binding derived from a reduced density of binding sites, rather than from affinity changes. The allosteric responsiveness of TBPS binding to exogenous GABA was also reduced in aged animals. These findings indicate an age-related change in the molecular (structural) organization of the GABAA receptor-chloride ionophore complex in rat cerebral cortex. PMID- 2545826 TI - A pathological study of the association between Lewy body disease and Alzheimer's disease. AB - The possibility of an association between Parkinson's disease and Alzheimer's disease has been examined by studying the age-specific prevalence of Lewy bodies in the substantia nigra in a group of 273 control cases without Parkinson's disease and 121 cases of Alzheimer's disease. The substantia nigra was also studied in 14 cases of Down's syndrome, 13 of which had cortical Alzheimer pathology. Twelve (7.8%) of the controls aged over 60 years showed nigral Lewy bodies. There was mild nerve cell degeneration and/or an extranigral distribution of Lewy bodies, suggestive of presymptomatic Parkinson's disease. Twenty five (22.5%) of the Alzheimer's disease cases over 60 years showed Lewy bodies, but only 14 (14.0%) of these had mild nigral cell loss consistent with presymptomatic Parkinson's disease. No case of Down's syndrome had Lewy bodies. Counts of tangles and plaques in hippocampus, frontal and temporal cortex were lower in cases of Alzheimer's disease with Lewy bodies compared with those without, but cortical choline acetyltransferase (ChAT) activities were similar. This suggests that Lewy body degeneration in the nucleus basalis of Meynert contributes to the deficit of cortical ChAT, but not to the cortical Alzheimer pathology. The relatively small difference in the prevalence of Lewy bodies between controls and Alzheimer's disease could be explained by the additive effects of Lewy body and tangle pathology causing dementia, rather than a greater than chance association between Parkinson's disease and Alzheimer's disease. PMID- 2545828 TI - Herpes simplex virus specific antibody determined by immunoblotting in cerebrospinal fluid of a patient with the Guillain-Barre syndrome. AB - The Guillain-Barre syndrome is often preceded by a herpes virus infection. Herpes simplex virus, however, has rarely been observed as the causative agent. A patient is described with a herpes simplex virus infection followed by a Guillain Barre syndrome. Immunoblotting was used to detect herpes simplex virus-specific antibodies in serum and cerebrospinal fluid. PMID- 2545827 TI - Diffuse Lewy body disease: clinical features in 15 cases. AB - Fifteen cases of diffuse Lewy body disease were diagnosed on pathological grounds during a single year in one health district. The range and frequency of clinical features contrast strikingly with previous reports. The majority of cases presented with classical levodopa-responsive Parkinson's disease either alone (6 cases) or with mild cognitive impairment (3 cases); the remaining 6 cases presented with cognitive impairment alone. In time almost all patients developed both dementia and Parkinsonism. The dementia was cortical in type, but unusual in that most (12 cases) showed day-to-day fluctuation in severity at some point in their illness. These findings suggest that diffuse Lewy body disease is not rare, and that it presents in a range of ways from dementia with subsequent Parkinsonism to Parkinson's disease with subsequent dementia. The latter mode of presentation suggests that it should be considered as a significant pathological substrate of dementia in Parkinson's disease. PMID- 2545829 TI - Hypergeusia as the presenting symptom of a posterior fossa lesion. PMID- 2545830 TI - The conduction velocities of peripheral nerve fibres conveying sensations of warming and cooling. PMID- 2545831 TI - Focal cytochrome c oxidase deficiency in various neuromuscular diseases. AB - To determine whether focal cytochrome c oxidase (CCO) deficiency characterized by scattered fibers with absent CCO activity among normal fibers was a specific finding for mitochondrial myopathies, we studied 389 muscle biopsies from various neuromuscular diseases other than mitochondrial myopathies. Focal CCO deficiency was found in 14 biopsies: 5 of 26 patients with myotonic dystrophy, 3 of 19 with nemaline myopathy, 1 of 7 with distal myopathy with rimmed vacuole formation, 3 of 22 with limb-girdle muscular dystrophy, 1 of 9 with amyotrophic lateral sclerosis, one of 79 with Duchenne muscular dystrophy. Focal CCO deficiency is known to be a crucial finding for chronic progressive external ophthalmoplegia, but it can also be seen in a variety of other neuromuscular disorders, probably as a secondarily induced phenomenon. PMID- 2545832 TI - Recombinant interferon-gamma in the treatment of recurrent nasopharyngeal carcinoma. AB - The etiologic relationship of the Epstein-Barr virus (EBV) to nasopharyngeal carcinoma prompted this study of the efficacy of interferon-gamma (IFN-gamma), which possesses antiviral and antitumor activity, in the treatment of recurrent carcinoma. Fourteen patients received recombinant IFN-gamma in daily intramuscular doses of 5-10 X 10(6) U/m2. Of 13 patients evaluable for response, five had a minor response; disease was stable in two and had progressed in six. EBV serology for immunoglobin (Ig) G, IgA-VCA, and early antigen was measured in six patients. In four of these, titers rose consistently with disease progression and in one patient, titers decreased with minor response. PMID- 2545833 TI - Selective elimination of axons extended by developing cortical neurons is dependent on regional locale: experiments utilizing fetal cortical transplants. AB - In adult rats, cortical neurons that extend an exon through the pyramidal tract (a major subcortical efferent projection of the neocortex) are limited to layer V of about the rostral two-thirds of the neocortex. In neonates, however, pyramidal tract neurons are distributed throughout the neocortex, but all of those found in certain areas, such as the posterior occipital region (including primary visual cortex) selectively lose their pyramidal tract axon (Stanfield et al., 1982) yet maintain axon collaterals to other subcortical targets (O'Leary and Stanfield, 1985). To determine if the regional location of a developing pyramidal tract neuron critically influences the maintenance or elimination of the axon collaterals it initially extends, pieces of cortex from embryonic day 17 (E17) rat fetuses (exposed to 3H-thymidine on E15) were transplanted heterotopically into the cortex of newborn (PO) rats; rostral cortex was placed into the posterior occipital region (R----O), or posterior occipital cortex into a rostral cortical locale (O----R). The retrograde tracers Fast blue (FB) and Diamidino yellow (DY) were used to assay for the presence of specific populations of cortical projection neurons within the autoradiographically identified transplants. In terms of the extension and maintenance of pyramidal tract axons, the transplanted neurons behave like the host neurons of the recipient cortical region rather than like those of their site of origin. At P40, following FB injections into the pyramidal decussation on P34, pyramidal tract neurons are labeled within the O----R transplants, but none can be labeled within R----O transplants, although in the same R----O cases transplanted neurons are labeled by an injection of DY in the superior colliculus. However, at P13 pyramidal tract neurons can be identified within the R----O transplants, as well as in the host occipital cortex, following injections made on P9, a period when the distribution of pyramidal tract neurons in normal rats is widespread (Stanfield and O'Leary, 1985b). In a second series of host rats, on P34 FB was injected in the pyramidal decussation of the O----R cases, or in the superior colliculus of the R----O cases, and in both groups DY was injected into the region of contralateral cortex homotopic for the new location of the transplant. On P40, in both the O----R and R----O transplants, many neurons singly labeled with FB or DY are found, but no double dye-labeled cells are seen.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2545834 TI - Fura-2 measurements of cultured rat Purkinje neurons show dendritic localization of Ca2+ influx. AB - The specific objectives of this study were the following: (1) to characterize the types of calcium currents in cultured PCs using whole-cell voltage-clamp techniques; (2) using fura-2 imaging techniques, to monitor intracellular Ca2+ levels during application of high potassium, glutamate, or glutamate analogs; and (3) to evaluate the types of calcium channels contributing to the calcium fluxes using pharmacological blocking agents. Voltage-clamp analysis of calcium currents proved to be difficult due to space-clamping problems. The latter was presumably due to the unfavorable geometry of cultured PCs. Nevertheless, we found no evidence for inward currents in cells bathed in TTX-TEA-BaCl2 saline. On the other hand, fura-2 measurements demonstrated that free Ca2+ levels were elevated in PCs following local application of either high-potassium saline or glutamate. When individual cells were injected with fura-2 and analyzed in TTX-containing saline, the Ca2+ elevation was usually greater in the dendrites. Since Ca2+ levels were not elevated in all dendrites of the same cell, the smaller responses in the soma wre not simply due to volumetric differences. Together with the voltage-clamp results, the fura-2 data indicate that calcium channels were localized to certain dendrites. Using selective calcium channel blockers, we found evidence for 2 types of calcium conductances in the dendrites of cultured PCs. The Ca conductance induced by high potassium was reduced in a dose-dependent manner by nifedipine (ED50 = 5 X 10(-7) M), indicating that a high-threshold voltage-dependent calcium channel was present. The Ca response to glutamate (or NMDA) was reduced by 2-amino-5-phosphonovaleric acid (ED50 = 10(-4) M), as well as by nifedipine or 10(-4) M LaCl3, indicating that both voltage-dependent and glutamate-coupled channels were opened by glutamate application. PMID- 2545835 TI - Spontaneous and beta-adrenergic receptor-mediated taurine release from astroglial cells are independent of manipulations of intracellular calcium. AB - Stimulation of beta-adrenergic receptors on LRM55 astroglial cells results in cAMP-dependent release of taurine. We have previously demonstrated that extracellular Ca2+ is not required for either spontaneous or receptor-mediated taurine release (Martin et al., 1988b). In the present series of experiments we investigated the relationship between changes in intracellular free Ca2+ ([Ca2+]i) and taurine release. [Ca2+]i was measured using the fluorescent probe fura-2 and was manipulated by changing the concentration of Ca2+ in the incubation medium and by using the Ca2+ ionophore ionomycin. [Ca2+]i was reduced from 150 +/- 95 nM (n = 46) in control medium (containing 1.1 mM CaCl2) to 46 +/- 10 nM (n = 43) in saline containing no CaCl2 and 10 microM EGTA. [Ca2+]i was rapidly elevated to greater than or equal to 1 microM in medium containing 100 microM CaCl2 and 10 microM ionomycin. Taurine release, either spontaneous or stimulated by isoproterenol, was not significantly affected by these manipulations of [Ca2+]i. [Ca2+]i did not change when cells were stimulated with 100 nM isoproterenol in either control saline containing 1.1 mM CaCl2 or in CaCl2 free saline containing 10 microM EGTA. Other secretogogs (serotonin and ethanol) did not cause changes in [Ca2+]i. These data indicate that neither spontaneous or receptor-mediated taurine release from astroglial cells is Ca2+ dependent. However, when cells were preloaded with Ca2+, allowed to recover briefly, and then stimulated with isoproterenol, it was possible to demonstrate transient increases in Ca2+.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545836 TI - Nasotemporal overlap of crossed and uncrossed retinal ganglion cell projections in the Japanese monkey (Macaca fuscata). AB - The nasotemporal overlap of crossed and uncrossed retinal ganglion cell projections were studied in 11 Japanese monkeys (Macaca fuscata) using HRP and fluorescent dyes (DAPI and RITC) as retrograde tracers and by physiological recordings of antidromic field potentials. A strip of nasotemporal overlap ran orthogonal to the horizontal meridian in all the whole-mount retinas studied. In HRP-labeled retinas of 6 monkeys, the width of the overlap gradually increased from 0.6 degrees in the central retina up to 5 degrees at eccentricity of 5 mm, and to 15 degrees at the extreme periphery. We also noted a clear asymmetric distribution of crossed and uncrossed retinal ganglion cell projections particularly in the perifoveal region; ipsilaterally projecting cells encircled the nasal edge of the fovea, whereas few contralaterally projecting cells were observed in the temporal edge. Soma-size analysis revealed that crossed projections in the temporal portion of the overlap arose mainly from large and small cells (presumably P alpha and P gamma cells, respectively); uncrossed projections in its nasal portion arose from medium-sized cells (presumably P beta cell). Direct evidence of the overlap as well as of the asymmetry was obtained in subsequent fluorescent dye experiments in 3 monkeys. Physiological studies on 2 additional monkeys confirmed the widening of the nasotemporal overlap towards the upper and lower parts of the retina. Moreover, in the nasal portion of the overlap, only slow potentials, which presumably reflect activities of P beta cells, were recorded after stimulation of the ipsilateral LGN as expected from the morphological study. The findings are discussed in relation to clinical observations of macular sparing and splitting, and with regard to the functional differences between P alpha and P beta cell systems on which binocular stereoscopic vision along the midsagittal plane may be based. PMID- 2545837 TI - Embryonic development of axon pathways in the Drosophila CNS. II. Behavior of pioneer growth cones. AB - We have identified the neurons that pioneer the major CNS axon tracts in the Drosophila embryo and determined their trajectory and fasciculation choices using serial section electron microscopy. Although Drosophila pioneer neurons make choices similar to those of their grasshopper homologs, there are interesting differences that reflect the much smaller nervous system size and the much faster rate of development characteristic of Drosophila. For example, where 2 longitudinal tracts are pioneered independently in grasshopper, only one is formed in Drosophila. This change is due to a change in fasciculation affinity of the pCC growth cone. Additionally, the intersegmental (IS) nerve is pioneered by a different neuron in Drosophila (aCC) than in the grasshopper (U1) because the smaller Drosophila CNS places the IS nerve within filopodial reach of the aCC soma, while in the grasshopper it is not. Drosophila growth cones explore a much more confined neuropil volume than do grasshopper growth cones but can also sample a larger fraction of the CNS as well. For this reason, some cell-cell recognition events critical to pathfinding in the grasshopper embryo may not be as essential in Drosophila. Nevertheless, many specific cellular affinities have been retained through the evolutionary divergence of these 2 species. PMID- 2545838 TI - Disuse-induced enhancement of Ia synaptic transmission in spinal motoneurons of the rat. AB - Monosynaptic excitatory postsynaptic potentials (EPSPs) evoked in rat spinal motoneurons by stimulation of a muscle nerve are enhanced during the first few days after section of the muscle nerve before subsiding to subnormal levels. We have examined whether this biphasic alteration results from changes of different factors involved in the regulation of central synaptic function. We chronically blocked the conduction of the intact peripheral nerve with tetrodotoxin (TTX). Under this disuse condition, enhancement of monosynaptic EPSPs was fully expressed within 2 d, and synaptic function remained elevated for at least 3 d, even after resumption of the impulse activity following discontinuance of the TTX. The magnitude of synaptic enhancement and the minimal period of nerve block required for synaptic enhancement were both comparable to those observed following nerve section. Thus, the early synaptic enhancement produced by section of the peripheral nerve could be accounted for entirely by the deprivation of sensory impulse activity. However, a prolonged nerve block up to 6 d still maintained synaptic enhancement. This was in contrast with synaptic depression observed after the first 3-4 d following section of the peripheral nerve. The late synaptic depression can thus be attributed to changes in some factors other than sensory impulse activity. Therefore, sensory impulse activity and the peripheral axonal continuity of sensory fibers appear to be 2 distinct factors for the regulation of central synaptic function. PMID- 2545839 TI - Voltage-dependent calcium channels regulate melatonin output from cultured chick pineal cells. AB - Chick pineal cells maintained in primary culture display a circadian rhythm of melatonin production and release, and the nocturnal increase in melatonin output is enhanced by elevating extracellular K+. The divalent cations, Co2+, Cd2+, and Mn2+, each reduce nocturnal melatonin output. Nitrendipine and nifedipine also prevent the nocturnal rise in melatonin output, while Bay K 8644 increases it, suggesting a role for voltage-dependent Ca2+ channels in regulating melatonin output. The whole-cell patch-clamp technique was used to record from individual chick pineal cells. Under conditions designed to isolate currents through voltage dependent Ca2+ channels, biphasic inward currents are elicited by large depolarizing commands (e.g., to 0 mV) from a holding potential of -90 mV; from a holding potential of -40 mV, only a sustained inward current is elicited by steps to 0 mV. Both components of the inward current are blocked by Co2+ or Cd2+. The sustained current is increased in amplitude by Bay K 8644 and blocked by nifedipine, while the transient current is unaffected. Since there is no evidence for vesicular release of melatonin, the "L-type" calcium channels mediating the sustained calcium current appear to be involved in the pathways regulating melatonin synthesis in chick pineal cells. PMID- 2545841 TI - Localization and characterization of melatonin receptors in rodent brain by in vitro autoradiography. AB - Little is known of the neural sites of action for the pineal hormone, melatonin. Thus, we developed an in vitro autoradiographic method using 125I-labeled melatonin (I-MEL) to study putative melatonin receptors in rodent brain. We first determined optimal in vitro labeling conditions for autoradiographic detection of I-MEL binding sites in rat median eminence, the most intensely labeled area in the rat brain. We then assessed the pharmacologic and kinetic properties of I-MEL binding sites in rat median eminence by quantitative autoradiography. These sites have high affinity for I-MEL (equilibrium dissociation constant = 43 pM). I-MEL binding was inhibited by nanomolar concentrations of melatonin or 6 chloromelatonin, but was not inhibited by serotonin, dopamine, or norepinephrine (100 microM). These results suggest that I-MEL binding sites identified by in vitro autoradiography represent specific, high-affinity melatonin receptors. Studies of the distribution of I-MEL binding in rat, Syrian hamster, and Djungarian hamster brain confirm that the median eminence and suprachiasmatic nucleus are major sites of I-MEL binding in rodent brain; other brain areas labeled in one or more of these species were the thalamus (paraventricular, anteroventral, and reuniens nuclei, nucleus of the stria medullaris, and medial part of the lateral habenular nucleus), hypothalamus (dorsomedial nucleus), subiculum, and area postrema. The presence of putative melatonin receptors in the suprachiasmatic nuclei and median eminence of these rodent species suggests that these brain regions are important loci for melatonin effects on circadian rhythms and reproduction. PMID- 2545840 TI - Release of vasoactive intestinal peptide in mouse cerebral cortex: evidence for a role of arachidonic acid metabolites. AB - In rodent cerebral cortex, vasoactive intestinal peptide (VIP) is contained in a homogeneous population of radially oriented bipolar interneurons. We have observed that 4-aminopyridine (4-AP), a K+-channel blocker, promotes a concentration- and Ca2+-dependent release of VIP from mouse cerebral cortical slices, with a significant effect already observed at 50 microM. Over 70% of VIP release elicited by 4-AP is blocked by 2 microM tetrodotoxin (TTX). Mepacrine, an inhibitor of phospholipase A2 (PLA2) activity and hence of arachidonic acid (AA) formation from membrane phospholipids, markedly inhibits (IC50 of approximately 15 microM) the release of VIP evoked by 4-AP. The inhibitory effect of mepacrine is not additive to that of TTX, thus indicating an involvement of PLA2 activation in the TTX-sensitive component of the 4-AP-evoked release. As a corollary, melittin (0.1-10 micrograms/ml), a PLA2 activator, promotes VIP release. Inhibition of AA metabolites of the lipoxygenase pathway by nordihydroguaiaretic acid, 5,8,11,14-eicosatetranoic acid, and caffeic acid results in a concentration dependent inhibition of VIP release evoked by 4-AP. This set of observations indicates for the first time that the formation of AA metabolites of the lipoxygenase pathway plays a role in the release of a peptide in the mammalian CNS. Furthermore, these observations together with the previously reported potentiation by prostaglandins of the increase in cyclic AMP elicited by VIP in mouse cerebral cortex (Schaad et al., 1987) indicate that AA metabolites may act at both the presynaptic (lipoxygenase metabolites) and the postsynaptic (cyclooxygenase metabolites) levels to increase the "throughput" or "strength" of VIP-containing circuits in the rodent neocortex. PMID- 2545842 TI - Inhibition of lymphocyte function by glioblastoma-derived transforming growth factor beta 2. AB - Human glioblastoma cells secrete an inhibitory factor termed "glioblastoma derived T-cell suppressor factor" (G-TsF). A member of the transforming growth factor beta (TGF beta) family, G-TsF is identical to TGF beta 2. The present study investigated the effect of G-TsF/TGF beta 2 on the proliferative and cytotoxic properties of tumor-infiltrating lymphocytes (TIL's) isolated from malignant gliomas after expansion in vitro with interleukin-2 (IL-2). The results demonstrate that the IL-2 (5 to 20 U/ml)-dependent proliferative response of glioma-derived TIL's was inhibited 70% to 85% by G-TsF/TGF beta 2 and that the inhibitory effect could be reduced by using increasing concentrations of IL-2 (100 to 200 U/ml). Tumor necrosis factor alpha (TNF alpha) enhanced the IL-2 dependent proliferation of TIL's cultured in low concentrations of IL-2 (10 U/ml); however, neither TNF alpha nor interferon gamma was able to reduce the inhibitory effect of TGF beta 2 on TIL proliferation. In addition, TGF beta 2 suppressed 60% to 100% the cytotoxic response of glioma-derived TIL's against several tumor targets, including autologous glioma cells, and the suppressive effect was shown to be reduced by increasing concentrations of IL-2. PMID- 2545843 TI - Quantification of the c-myc oncoprotein in human glioblastoma cells and tumor tissue. AB - The identification of a quantifiable oncoprotein marker in glial cells could lead to its use as an aid in the diagnosis, grading, and treatment of tumours of glial origin. In this study, monoclonal antibodies to the c-myc oncoprotein were used in conjunction with immunofluorescence microscopy, flow cytometry, and immunoblot analysis to quantitate and characterize the expression of this oncoprotein in neoplastic and benign cultured glial cells and brain-tumor tissue. Flow cytometric analysis revealed that the c-myc oncoprotein was highly expressed in neoplastic cell lines and in glioblastoma tumor specimens. In contrast, anti-c myc oncoprotein staining was not present in a non-neoplastic glial cell line or in a benign brain tissue specimen. Immunoblot analysis revealed two distinct c myc oncoprotein bands, having molecular weights of 64 and 75 kD. Densitometric determinations of the relative levels of the 64-kD protein were in good agreement with the determinations made by flow cytometry. Flow cytometry was also used to relate the quantity of the c-myc oncoprotein present in the cells to their cell cycle phase. In the malignant cultured cells, the protein underwent an approximate twofold increase as the cells progressed from G1/G0 to G2/M in the cell cycle. The present results suggest that the c-myc oncoprotein may prove to be a useful marker for the proliferation status and/or malignancy of glial cells. PMID- 2545844 TI - Demonstration of beta 1-adrenergic receptors in human placenta by ( )[125I]iodocyanopindolol binding. AB - The highly specific beta-adrenergic radioligand (-)[125I]iodocyanopindolol (ICYP) was used to characterize the beta-adrenergic receptor subtype present in human placenta. Binding of ICYP to membranes from human placenta was saturable with time and ligand concentration, of high affinity, and demonstrated appropriate stereoselectivity and agonist rank order of potency for binding to a beta adrenergic receptor. From saturation binding curves, the KD and Bmax values for ICYP binding were 233 +/- 51 pM and 690 +/- 139 fmol/mg of proteins, respectively. Analysis of inhibition of ICYP binding by beta 1- and beta 2 selective adrenergic antagonists via Hofstee analysis resulted in linear plots, indicating the existence of a homogeneous population of beta-adrenergic receptors. From the resulting KI-values for the beta 1-selective drugs practolol (4.0 +/- 0.9 microM) and metoprolol (0.19 +/- 0.07 microM) and for the beta 2 selective drug ICI 118,551 (0.30 +/- 0.06 microM) it is concluded that the beta adrenergic receptor in human placenta is of the beta 1-subtype. This is further supported by the fact that (-)-noradrenaline and (-)-adrenaline were equipotent in inhibiting ICYP binding. PMID- 2545846 TI - Effects of dietary fiber and protein concentration on growth, feed efficiency, visceral organ weights and large intestine microbial populations of swine. AB - Finishing barrows (average initial weight 55.5 +/- 2.4 kg) were used to determine the effects of high dietary fiber or protein on performance, visceral organ weights and large intestine microbial populations and to monitor the duration of regression of swine visceral organ mass and microbial populations to control values following transfer from the high fiber or high protein diet to the control diet. Four pigs from each diet were killed on d 17, 34, 48 and 66. From d 34 until slaughter 14 and 32 d later, all remaining pigs were fed the control diet ad libitum. High fiber resulted in significantly higher relative weight of the total gastrointestinal tract after 34 d and higher relative stomach weight up to d 48. Compared with the control diet, the high protein diet resulted in increased relative liver and kidney weights up to d 48. The number of proteolytic and cellulolytic bacteria increased in the colon contents of pigs fed a high protein or high fiber diet, respectively, but declined to below control values within 14 d of transferring pigs from the high protein or high fiber diet to the control diet. The results indicate that diet composition plays a more specific role in visceral organ hypertrophy than can be explained by the normal relative changes in organ size as body weight increases. Thus, high dietary fiber and protein may indirectly increase the animal's maintenance requirement by causing a repartitioning of nutrients from the edible carcass to the visceral organs. PMID- 2545845 TI - Influence of cellulose structure on its digestibility in the rat. AB - The effect of modifying the structural properties of a purified cellulose on its subsequent digestion in the rat was determined. The three structurally unique types of cellulose used in the study were a commercial microcrystalline cellulose, a ball-milled cellulose, and an acid-swollen cellulose. The ball milled cellulose and the acid-swollen cellulose were prepared from the microcrystalline cellulose starting material. Differences in structure between the types of cellulose were determined by X-ray diffraction measurements and by analysis of their in vitro enzymatic saccharification. The extent of fermentation of each type of cellulose within the rat intestinal tract was determined. Each of the three types of cellulose had a unique structure with respect to its measured crystallinity index and its rate of degradation in vitro by cellulase enzymes. The measured in vivo digestion coefficient for microcrystalline, ball-milled, and acid-swollen cellulose was 8.8, 12.2 and 20.3%, respectively. This represents a significant difference (P less than 0.05) in the extent of fermentation of the different structural types of cellulose within the rat intestine. The results demonstrate that modifying the structural properties of dietary cellulose can affect in vivo response. PMID- 2545847 TI - Gas-producing necrotizing fasciitis following mandibular fracture. AB - A case of necrotizing fasciitis following a mandibular fracture is reported. The diagnosis and treatment of the disease are discussed. PMID- 2545848 TI - Dentine hypersensitivity. II. Effects produced by the uptake in vitro of toothpastes onto dentine. AB - Toothpastes arguably are the most common preparations used in the treatment of dentine hypersensitivity. Evidence for efficacy largely comes from clinical trials, and supports the role of the contained so-called "active" ingredient. How such compounds produce therapeutic benefit has received limited attention. This study measured the availability and uptake onto dentine of fluoride and metal ions contained in commercial and test toothpastes. The effects of exposure of dentine sections to these toothpastes, and abrasive only pastes in particular, was investigated by scanning electron microscopy and X-ray microanalysis. Aqueous extracts of the different toothpastes contained measurable concentrations of the incorporated metal and or fluoride, but usually considerably below that incorporated by the manufacturer. Extraction of dentine sections exposed to toothpaste slurries revealed levels of fluoride and metal ions, although post treatment washing in water reduced these levels. Surface changes at 1 and 10 h were not consistent with the uptake of fluoride or metals. These changes were almost certainly produced by the contained abrasives, including calcium carbonate, dicalcium phosphate, alumina and silica. In particular fume silica progressively accumulated on the dentine surface to narrow and occlude dentinal tubules. Unlike other abrasives fume silica was resistant to removal by washing. It would appear that insufficient attention has been given to the therapeutic potential of toothpaste ingredients, particularly abrasives, to produce benefit by occlusion of dentinal tubules. PMID- 2545849 TI - [The relationship between the number of the autonomic nerve receptors and degree of hyperreactive nasal symptoms in patients with hyperesthetic rhinitis]. AB - It is generally accepted that abnormal autonomic responsiveness may contribute to the pathogenesis of hyperesthetic rhinitis. Histologically, in the nasal mucosa, cholinergic fibers are found close to blood vessels, but are particularly numerous around the glands. Adrenergic fibers are found mainly around the vascular structures. Physiological and pharmacological studies demonstrate that parasympathetic hypersensitivity causes hypersecretion, and sympathetic hyposensitivity causes vasodilatation. alpha 1-adrenergic receptor function is dominant for this vasodilatation. Using radioligand binding techniques, it has been found that there is an increased number of muscarinic cholinergic receptors and a decreased number of alpha 1- and beta-adrenergic receptors in patients with nasal allergy, while the binding affinities do not change. In this report, using radioligand binding techniques, we investigated the relationships between the number of receptors and the degree of the hyperreactive nasal symptoms in patients with hyperesthetic rhinitis. The results are as follows. 1. The number of muscarinic cholinergic receptors of human nasal mucosa in patients with hyperesthetic rhinitis was related significantly (P less than 0.01) to the degree of hypersecretion induced by methacholine and frequency of blowing nose estimated from allergy diary. 2. There was no relationship between frequency of sneezing and the number of muscarinic cholinergic receptors. 3. The number of alpha 1 adrenergic receptors was related significantly (P less than 0.05) to the degree of swelling of nasal mucosa induced by methoxyamine. Judging from these results, it was assumed that pathogenesis of hyperreactive nasal symptoms may be associated at least partially to the changes of number of autonomic nerve receptors in the nasal mucosa. PMID- 2545851 TI - [Case report of catecholamine-secreting carotid body tumor]. AB - We encountered a case of catecholamine-secreting carotid body tumor which has never been reported in Japan. The patient was a 20-year old man, who complained of a mass located at the right neck and hypertensive symptoms such as loss of consciousness at extension of the neck, fatigability, etc. Carotid angiography and computed tomography (CT scan) revealed a carotid body tumor. Before operation, a mild hypertension was noted, serum and urinary levels of norepinephrine showed abnormal increase, and phentolamine test was positive. After the tumor was dissected by a surgical procedure associated with carotid reconstruction, blood pressure and norepinephrine levels clearly trended to decrease and phentolamine test returned to negative. Histopathologically, the tumor was a paraganglioma with so-called "organoid" pattern. The tumor was not chromaffin, but the presence of neurosecretory granules was confirmed under electron microscope. In one-year follow up after the operation, no sign suggestive of "latent" or newly arising paraganglioma was noted in assay of catecholamines in serum and urine, nor on adrenal CT scan. PMID- 2545850 TI - [Suppressive effect of CDDP on induction of the Epstein-Barr virus antigen synthesis by n-butyrate]. AB - n-Butyrate has been shown to induce Epstein-Barr virus (EBV) antigen synthesis in certain EBV genome-carrying lymphoblastoid cell lines. We have studied the effect of cis-dichloro-diamine-platinum (CDDP) on induction of EBV antigen synthesis not only by n-butyrate treatment of the EBV producer, P3HR-1 cells, but also by superinfection of EBV to the EBV non-producer, Raji cells. Simultaneous treatment of the cells with n-butyrate and CDDP or MMC blocked VCA and EA induction, but no suppressive effect was observed by the treatment of the cells with n-butyrate and Ara-C. The induction of EBV antigens was also observed in EBV-superinfected Raji cells, even in the presence of DNA synthesis inhibitor, CDDP or MMC. From these results, we discussed the mechanisms of the EBV antigens synthesis in EBV latently infected cells. PMID- 2545852 TI - A comparison of fluconazole and ketoconazole in the treatment of rat palatal candidosis. AB - The efficacy of fluconazole and ketoconazole in the treatment of rat palatal candidosis has been investigated. To induce oral candidosis, the palatal tissues of Wistar rats were inoculated with Candida albicans NCPF 3091 and covered with acrylic plates. The rats were treated with either fluconazole or ketoconazole by intragastric gavage for 14 days. Palatal epithelial thickness, the number of yeasts present and the histopathological appearance of the tissue were assessed to compare treatment with the two azoles. A fluconazole dose of 0.75 mg kg-1 body weight once daily for 14 days was required to cure the palatal candidosis and prevent recrudescence, whereas with ketoconazole a dose of 7.0 mg kg-1 body weight was necessary to achieve the same effect. From these results it is concluded that fluconazole is effective at a dose nine times lower than ketoconazole in resolving rat palatal candidosis. PMID- 2545853 TI - Alteration in bone metabolism with increasing age: effects of zinc and vitamin D3 in aged rats. AB - The alteration in bone metabolism with increasing age was investigated in the femoral diaphysis of male rats. Calcium content was highest in the bone from 3 week-old rats (491 +/- 13 mg/g bone ash), falling gradually with aged to 357 +/- 7 and 306 +/- 9 mg/g bone ash in 28- and 52-week-old rats, respectively. Bone zinc content increased until rats were 3 weeks of age, and thereafter remained constant. Deoxyribonucleic acid (DNA) content was highest in the bone from 1-week old rats, and it decreased markedly with increasing age. Alkaline phosphatase and acid phosphatase activities increased up to 3 weeks, then subsequently declined with age. Thus, the retardation of bone metabolism was induced by ageing. When zinc sulfate (5.0, 10.0 and 20.0 mg Zn/kg body weight) was administered orally for 3 d to 28-week-old rats, alkaline phosphatase activity and calcium content in the femoral diaphysis was elevated markedly by all doses. The oral administration of vitamin D3 (2.0 and 20 micrograms/kg) for 3 d in 28-week-old rats did not produce an appreciable increase in bone alkaline phosphatase activity or calcium content, while 1,25-dihydroxyvitamin D3 (1.5 micrograms/kg) caused a significant increase in those biochemical indices. These results indicate that zinc and 1,25 dihydroxyvitamin D3 play a role as activators in bone metabolism of ageing rats. PMID- 2545854 TI - Stimulatory and inhibitory effects of forskolin on adenylate cyclase in rat normal hepatocytes and hepatoma cells. AB - Forskolin synergistically potentiated adenosine 3',5'-cyclic monophosphate formation by prostaglandin E1 (PGE1) in rat normal hepatocytes freshly prepared by collagenase digestion and rat ascites hepatoma AH66 cells, but dose dependently inhibited the accumulation by PGE1 in AH66F cells. Forskolin activated adenylate cyclase in a dose-dependent manner in homogenates of all cell lines. In normal hepatocytes and AH66 cells, simultaneous addition of forskolin and other adenylate cyclase activators [isoproterenol (IPN), PGE1, guanosine 5' triphosphate sodium salt (GTP), 5'-guanylylimidodiphosphate sodium salt (Gpp (NH)p), NaF, cholera toxin, islet activating protein and MnCl2] gave greater than additive responses. On the other hand, in AH66F cells, the effect of forskolin on adenylate cyclase was hardly influenced by GTP, but forskolin diminished the activities induced by high concentrations of GTP to that by the diterpene alone. Forskolin also significantly inhibited the PGE1-stimulated and the guanine nucleotide binding regulatory protein-stimulated activities. Because AH66F cells were insensitive to IPN, the combination with forskolin and IPN gave similar activity to that obtained with the diterpene alone. The effect of forskolin on the activation by manganese ion was neither synergistic nor inhibitory but was additive in AH66F cells. These results suggest that forskolin promotes the interaction between the stimulatory guanine nucleotide binding regulatory protein and the catalytic unit in normal hepatocytes and AH66 cells, but in AH66F cells forskolin interferes with the coupling of the two components of adenylate cyclase. PMID- 2545855 TI - [Function of protein myristoylation in cellular regulation and viral proliferation]. AB - Protein myristoylation was first discovered in the catalytic subunit of adenosine 3',5'-cyclic monophosphate-dependent protein kinase. Subsequently, various cellular and viral myristoylated proteins were detected. In each case, the myristoyl moiety was found in an amide linkage with the amino terminal glycine residue of the modified proteins. The biological functions of protein myristoylation of various cellular protein, oncogene product, and viral structural proteins have been studied by many biochemists. Two of the most thoroughly studies myristoylated proteins are the transforming protein of Rous sarcoma virus, pp60v-src, and the proto-oncogene product, pp60c-src. Deletion, modification of the first 14 NH2-terminal amino acid of pp60v-src, or chemical antimyristoylation of the protein with N-myristoyl glycinal diethylacetal does not affect intrinsic tyrosine src-kinase activity, but prevents myristoylation and membrane association, and abolishes the transforming activity of the protein. Protein myristoylations of some viral structural proteins were also studied by many investigators, and X-ray crystallographic studies of poliovirus suggest that myristate moiety may play a central role in capsid assembly. Recently, human immunodeficiency virus, HIV-I, process a myristoylated p17gag protein, which is proteolytically derived from the NH2-terminus of a gag precursor protein, and its myristate moiety may be important for virus assembly. In this review, we detailed recent studies of the protein myristoylation in cellular regulation and virus proliferation. PMID- 2545856 TI - Pharmacological profile of NPC 12626, a novel, competitive N-methyl-D-aspartate receptor antagonist. AB - The novel compound 2-amino-4,5-(1,2-cyclohexyl)-7-phosphonoheptanoic acid (NPC 12626) was evaluated for activity in a variety of tests associated with receptors for excitatory amino acids. NPC 12626 failed to inhibit the specific binding of RS-[3H] amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid or [3H] kainic acid to brain membranes in vitro but displaced both agonist and antagonist binding to N-methyl-D-aspartic acid (NMDA) receptors. Like cis-(+/-)-3-(2-carboxypiperazine 4-yl)propyl-1-phosphonic acid, NPC 12626 competitively blocked NMDA-induced enhancement of [3H]-1-thienylcyclohexyl)piperidine binding. In the voltage clamped frog oocyte expression system, NPC 12626 was a competitive inhibitor of NMDA-evoked inward current with a pA2 of 6.24. After both i.c.v. or i.p. administration, NPC 12626 was a potent anticonvulsant in the pentylenetetrazol, maximal electroshock and NMDA seizure models. Furthermore, low doses (25 mg/kg) of NPC 12626 given i.v. were effective in preventing damage to the CA1 region of hippocampus in the gerbil model of global ischemia. Unlike the noncompetitive NMDA antagonist, phencyclidine, but like cis-(+/-)-3-(2-carboxypiperazine-4 yl)propyl-1-phosphonic acid and pentobarbital, NPC 12626 only partially substituted for phencyclidine in a drug discrimination study. The results of the current study indicate that NPC 12626 is a novel, systemically active and competitive NMDA receptor antagonist. PMID- 2545857 TI - Guanine nucleotides are competitive inhibitors of N-methyl-D-aspartate at its receptor site both in vitro and in vivo. AB - Guanine nucleotides were shown to alter N-methyl-d-aspartate (NMDA) receptor effector coupling by competitive antagonism at the glutamate binding site, rather than via interaction with an intracellularly located GTP-binding protein. Thus, in contrast to known G-protein linked receptors, micromolar concentrations of guanine nucleotides and their analogs decreased both agonist [( 3H]glutamate) and antagonist [( 3H]-3-(2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid binding to the NMDA receptor complex. The most potent compound, the GDP analog guanosine-5' O-(2-thiodiphosphate) (GDP beta S), was studied in detail. GDP beta S exhibited almost 200-fold selectivity for the glutamate recognition site vs. the strychnine insensitive glycine binding site. IC50 values were 2.7 +/- 1.4 and 484 +/- 97 microM, respectively. GDP beta S also inhibited N-[1-(2-thienyl)cyclohexyl 3H]piperidine binding (IC50 was 28.0 +/- 3.7 microM) in an NMDA-reversible fashion. [3H]-3-(2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid saturation binding studies revealed an increase in Kd from 263 +/- 49 (control) to 552 +/- 134 nM (8 microM GDP beta S) without any change in maximum binding (4.94 +/- 0.34 and 5.19 +/- 0.58 pmol/mg of protein, respectively). GDP beta S was also a competitive inhibitor of the following NMDA-stimulated responses: elevation of cyclic GMP in neonatal rat cerebellar slices, release of preloaded [3H]norepinephrine from superfused rat hippocampal slices and elevation of cytosolic calcium concentration in fura-2-loaded cultured rat forebrain neurons. IC50 values were 78.4, 53.4 and 1.6 microM, respectively. Finally, GDP beta S resembled known NMDA receptor antagonists in its ability to block NMDA receptor induced seizures after i.c.v. administration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545858 TI - Intraperitoneal endotoxin but not protein malnutrition shifts d-tubocurarine dose response curves in mouse gastrocnemius muscle. AB - Burn injury-induced changes at the neuromuscular junction include muscle weakness and altered response to neuromuscular blocking drugs. Protein malnutrition and sepsis can concomitantly occur with burn trauma. The role of pure malnutrition or sepsis, in the absence of burn injury, in inducing neuromuscular changes was studied in the mouse gastrocnemius muscle. Additionally, cAMP levels in muscle were evaluated to reflect metabolic activity. Sepsis was studied using doses of endotoxin at one-fourth or one-third the dose evoking 50% lethality. Diets of 5% protein and 5% protein + 35% fiber achieved protein and protein/calorie malnutrition, respectively. In each model neuromuscular function was evaluated by maximal tension developed. Pharmacologic responses were measured through effective dose to paralyze active tension by either 50 or 95%. Protein and protein/calorie malnutrition leading to an approximate 8% body weight loss caused a depression of maximal tension developed; this depression in tension was associated with a 10-fold increase in cAMP levels. Effective doses of d tubocurarine for twitch inhibition during malnutrition were not significantly different from controls. Sepsis at 2 weeks caused an approximate 8% body weight loss, a significant decrease in maximal tension and at least a 3- to 5-fold shift to the right in dose-response curves to d-tubocurarine. In contrast to malnutrition, cAMP levels were significantly decreased (P less than .001) in sepsis to 1/400 of controls. The altered neuromuscular function and pharmacology observed in sepsis are similar to changes observed in burn injury. Protein malnutrition common to these two states may be important in functional but not pharmacological changes at the neuromuscular junction. PMID- 2545859 TI - Phosphatidic acid phosphatase in neonatal rat lung: effects of prenatal dexamethasone or terbutaline treatment on basal activity and on responsiveness to beta adrenergic stimulation. AB - Phosphatidic acid phosphatase (PAPase) is a key enzyme in the synthesis of lung surfactant. This study compares the effects of prenatal exposure (gestational days 17, 18 and 19) to two drugs which enhance surfactant synthesis: dexamethasone (0.2 mg/kg) and terbutaline (2 or 10 mg/kg). Maternal dexamethasone treatment did not cause an initial stimulation of lung PAPase but did eventually evoke a small increase after the 1st postnatal week. The effect was selective in that brain PAPase activity was generally unaffected; liver PAPase was stimulated during the early neonatal period only. Dexamethasone also prolonged the developmental period of peak reactivity of lung PAPase to beta developmental period of peak reactivity of lung PAPase to beta adrenergic stimulation (tested with acute isoproterenol challenge), which ordinarily accompanies genesis of alveoli in the 2nd to 3rd postnatal week. Significant growth retardation was present even at this low dose of dexamethasone. In contrast, maternal administration of the beta adrenergic agonist, terbutaline, resulted in a large increase in basal enzyme activity in the lung during the immediate perinatal period and enhanced the responsiveness to isoproterenol challenge. The effect of terbutaline was accompanied by little or no growth impairment. Thus, although prenatal administration of either glucocorticoids or beta adrenergic agonists can enhance lung PAPase activity and reactivity to stimulation, the two classes of drugs differ substantially in time course of effect and in the propensity to retard growth. PMID- 2545860 TI - Potent stimulation of myofilament force and adenosine triphosphatase activity of canine cardiac muscle through a direct enhancement of troponin C Ca++ binding by MCI-154, a novel cardiotonic agent. AB - In the present study we have analyzed a likely biochemical mechanism underlying the Ca++-sensitizing action of MCI-154 (6-[4-(4'-pyridyl)aminophenyl)-4,5-dihydro 3(2H)-pyridazinone hydrochloride), a novel cardiotonic agent, on the contractile protein system. MCI-154 (10(-7) to 10(-4) M) enhanced the tension development induced by -log molar-free Ca++ concentration (pCa) 5.8 in chemically skinned fiber from the canine right ventricular muscle in a concentration-dependent manner. At pCa 7.0, MCI-154 (10(-7) to 10(-4) M) markedly increased adenosine triphosphatase (ATPase) activities of canine myofibrils and reconstituted actomyosin. In myofibrils and reconstituted actomyosin, MCI-154 (10(-7) to 10(-4) M) caused a parallel shift of the pCa-ATPase activity relation curve to the left without affecting the maximum activity, suggesting an increase in Ca++ sensitivity. MCI-154 (10(-8) to 10(-4) M) had little effect on actin-activated, Mg++, Ca++ and (K+, EDTA)-ATPase activities of myosin. Ca++ binding to cardiac myofibrils or purified cardiac troponin was increased by 10(-4) M MCI-154. These results suggest that MCI-154 enhances Ca++ binding to cardiac troponin C to elevate the Ca++ sensitivity of myofilaments and thus may cause a positive inotropic action in cardiac muscle. MCI-154 may provide a valuable tool for studying the molecular mechanism by which Ca++ regulates the contractile system. PMID- 2545861 TI - Evidence for the existence and ionic modulation of platelet-activating factor receptors mediating degranulatory responses in human polymorphonuclear leukocytes. AB - High-affinity binding sites for platelet-activating-factor (PAF) have been identified in human polymorphonuclear leukocytes (PMNs). The aim of this investigation was to assess their functional relevance by characterizing PAF induced elastase release, an enzyme present in azurophilic granules. At 37 degrees C, maximal release (measured with a spectrofluorimetric method) was achieved with 0.3 microM PAF. PAF analogs (e.g. lyso-PAF, 2-O-methyl-lyso-PAF) and enantio-PAF, which exhibit poor PAF-like activity in binding studies, were very weak releasers of elastase. The degranulation induced by PAF was fully and competitively inhibited by RP 59227, a potent, d-enantiomeric PAF antagonist. This effect was stereoselective and specific inasmuch as this compound was 500 fold more potent than its l-enantiomer and was devoid of effects on the degranulation promoted by N-formyl-l-methionyl-l-leucine-l-phenylalanine or [5S,12R]hydroxyeicosa-6,14-cis-8,10-trans-tetraenoic acid. The PAF-induced elastase extrusion was enhanced by physiological concentrations of CaCl2 and MgCl2. For instance, 1 mM Ca++ increased by 200% the release produced by 100 nM PAF. The mechanism of this effect is probably related to the capability of Ca++ and Mg++ to enhance the maximum number of [3H]PAF binding sites in whole or lysed PMNs without affecting the affinity (Kd) of the ligand. An additional possibility is that these ions intervene at the level of the signal transduction system leading to degranulation. For a series of 22 known PAF receptor antagonists, belonging to different chemical families, there was a strong correlation (r = 0.95) between their ability to displace [3H]PAF and to inhibit the PAF-evoked elastase release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545862 TI - Relationship between biotransformation of glyceryl trinitrate and cyclic GMP accumulation in various cultured cell lines. AB - We assessed glyceryl trinitrate (GTN) biotransformation and cyclic GMP accumulation in cultured rat lung fibroblasts (RLF), porcine kidney epithelial (PK1), bovine aortic endothelial (BAE) and bovine aortic smooth muscle (BASM) cells. Biotransformation of 0.1 microM GTN was linear over 30 min and the percentage of glyceryl dinitrate (GDN)/10(6) cells for BAE, BASM, RLF and PK1 at 30 min was 3.1, 2.3, 5.8 and 21.7%, respectively. At low GTN concentration (0.01 0.1 microM) there was a highly selective formation of 1,2-GDN, whereas at higher GTN concentration (greater than 1 microM) this selectivity was lost. Cyclic GMP accumulation did not occur in BAE or BASM at any GTN concentration, whereas for RLF and PK1 it was highly correlated to the rate of GDN formation. Upon re exposure to GTN after treatment of RLF or PK1 cells for 3 hr with 0.1 mM GTN, there was an almost complete loss of the cyclic GMP response, GTN biotransformation was attenuated markedly and the selective formation of 1,2-GDN at low GTN concentration was absent. However, when GTN-treated cells were incubated for 18 hr in GTN-free media, there was a recovery of the cyclic GMP response, GTN biotransformation and selective 1,2-GDN formation toward control values.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545863 TI - Behavioral evidence for the role of noradrenaline in the putative anxiogenic actions of the inverse benzodiazepine receptor agonist methyl-4-ethyl-6,7 dimethoxy-beta-carboline-3-carboxylate. AB - The role of noradrenaline in the anxiogenic action of the inverse benzodiazepine receptor agonist methyl-4-ethyl-6,7-dimethoxy-beta-carboline-3-carboxylate (DMCM) was assessed in a drug discrimination procedure using yohimbine as the stimulus cue and a conflict procedure using a conditioned behavioral suppression paradigm in rats. The yohimbine cue was antagonized by clonidine (0.03-0.5 mg/kg) and diazepam (0.1-10.0 mg/kg). DMCM (0.1-0.7 mg/kg) only partially substituted for the yohimbine stimulus cue. However, DMCM, in a smaller dose (0.1 mg/kg), significantly shifted the dose-effect curve of the yohimbine cue to the left. This potentiating effect of DMCM on the yohimbine cue was antagonized by Ro 15 1788, a benzodiazepine receptor antagonist, and by the type I benzodiazepine receptor agonist CL218,872. In the conditioned behavioral suppression paradigm, both clonidine (0.03 mg/kg) and diazepam (3 mg/kg) had an anticonflict effect by increasing responses in the conditioned fear period, whereas DMCM (0.1, 0.5 mg/kg) decreased the responses of rats in the conditioned fear period. This proconflict effect of DMCM was antagonized by muscimol (0.5 mg/kg), a type A gamma-aminobutyric acid receptor agonist, Ro 15-1788 (3 mg/kg) and clonidine (0.01 mg/kg). Our results suggest that the depressive effect of DMCM on the function of the gamma-aminobutyric acid-benzodiazepine receptor complex may cause increased noradrenergic activity, which may, in turn, be one of the anxiogenic mechanisms in DMCM. PMID- 2545864 TI - Alpha adrenoceptor stimulation reduces outward currents in rat ventricular myocytes. AB - The effects of alpha adrenoceptor stimulation with noradrenaline were investigated in rat ventricular myocytes after blockade of beta receptors with propranolol (1 microM). At room temperature and low stimulation frequency (0.5 Hz), noradrenaline evoked a phentolamine-sensitive increase in contraction amplitude by 22%. The action potentials of myocytes were prolonged. When the sodium current was inactivated by depolarization in whole-cell voltage clamp experiments, noradrenaline caused a small, but highly variable increase in net inward current and shifted the current-voltage relation between -30 and -5 mV to the hyperpolarizing direction. These effects were absent when K+ currents were inhibited by Cs+ substitution. After inhibition of the Ca++ current with Cd++ (0.1 mM), noradrenaline decreased the peak transient outward current; it reduced the steady-state outward current in a concentration-dependent manner (pD2 value, 6.9), but had no effect on the amplitude of the transient component of outward current. Noradrenaline reduced holding current at -40 mV. The inward branch of the inward rectifier was not affected. The noradrenaline-induced changes in membrane currents were significantly smaller in the presence of phentolamine (10 microM). They are therefore considered to be mediated by alpha adrenoceptor stimulation. The reduction in outward currents can explain the prolongation in action potential duration which could contribute to the increase in contractility. PMID- 2545865 TI - Evidence for two populations of 5-hydroxytryptamine receptors in dog basilar artery. AB - The aim of the present study was the classification of the receptors that mediate 5-hydroxytryptamine (5-HT)-induced responses in dog basilar artery. Isolated preparations from basilar artery of mongrel dogs denuded of endothelium were contracted by 5-HT in the presence of 6 microM cocaine. In the presence of either ketanserin or spiperone, concentration-effect curves for 5-5-HT became biphasic. The responses to low concentrations of 5-5-HT were resistant to blockade by either antagonist. The responses to high concentrations of 5-HT were antagonized by ketanserin and spiperone in a concentration-dependent and partially surmountable manner. The lack of complete surmountability was at least partially due to fade during the determination of the cumulative concentration-effect curves. The pKB values for the component of the 5-HT-induced contractions that was antagonized by ketanserin and spiperone were 9.4 and 10.2 (-log M), respectively. The findings are consistent with the assumption of an interaction of ketanserin and spiperone with 5-HT at 5-HT2 receptors. On the other hand, the response to low concentrations of 5-HT is not mediated through 5-HT2 receptors. This response is antagonized by phentolamine with an affinity approximately 10 times lower than its affinity to 5-HT2 receptors, but not by prazosin or benextramine. It is conceivable that the receptor that mediates the response to low concentrations of 5-HT belongs to the 5-HT1A-subpopulation as suggested recently. PMID- 2545866 TI - Calcium-independent activation of contractile apparatus in smooth muscle by calyculin-A. AB - Calyculin-A (CL-A), a novel marine toxin isolated from Discodermia calyx, caused contraction in the smooth muscle of guinea pig taenia ceci and rat aorta in the presence or absence (with 1 mM ethylene glycol bis(beta-aminoethyl ether)-N,N' tetraacetic acid) of external Ca++ at concentrations ranging from 1 X 10(-8) to 1 X 10(-6) M. In the presence of external Ca++, the contraction induced by CL-A was accompanied by an increase in the cytosolic free Ca++ concentration [( Ca++]cyt) as measured by the fluorescence indicator fura-2. Verapamil (3 X 10(-6) M) inhibited the increase in [Ca++]cyt, but not tension development caused by CL-A. In the absence of external Ca++, CL-A still caused contraction without changing [Ca++]cyt. Thus, from studies with intact smooth muscle it was demonstrated that, in the absence of external Ca++, CL-A can induce a contraction that was not accompanied by an increase in [Ca++]cyt. In permeabilized taenia, CL-A caused contraction in the absence of Ca++ (with 2 mM ethylene glycol bis(beta-aminoethyl ether)N,N'-tetraacetic acid) at concentrations similar to those required to contract intact tissue. This contraction was inhibited by the nonselective kinase inhibitors such as amiloride (1 X 10(-3) M) and K-252a (2 X 10(-5) M). Low concentrations of Ca++ (approximately 1 X 10(-6) M) augmented the CL-A-induced contraction in the permeabilized taenia. In native actomyosin prepared from chicken gizzard CL-A induced phosphorylation of the 20 kDa myosin light chain (MLC) in the absence of Ca++.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545867 TI - Evidence that prostaglandins can contract the rat aorta via a novel protein kinase C-dependent mechanism. AB - The effects of prostaglandin (PG) F2 alpha and PGB2 on isolated rat aortic strips were studied in calcium-free 1 mM ethylene glycol bis(beta-aminoethyl ether)-N,N' tetraacetic acid buffer to explore the mechanisms of PG-induced smooth muscle contraction. In the absence of extracellular calcium, PGF2 alpha and PGB2 induced sustained contractions that were similar to those induced by activators of protein kinase C (PKC) or norepinephrine. These sustained contractions were apparently induced via a pharmacomechanical coupling mechanism because they could be elicited repeatedly in the absence of extracellular calcium and were not affected by changes in buffer concentrations of potassium, magnesium or phosphate. They depended on intracellular but not extracellular calcium because they were reversed by nitroprusside but not by nifedipine, verapamil or diltiazem. Contractions induced by combining either PG with 12-O-tetradecanoyl phorbol-13-acetate or mezerein were consistent with the suggestion that PG induced contractions were induced via a PKC-dependent mechanism. Moreover, both PGF2 alpha- and PGB2-induced contractions were inhibited noncompetitively by 1-(5 isoquinolinylsulfonyl)-2-methylpiperazine at concentrations known to inhibit PKC but not myosin light chain kinase. These data, combined with those published from other laboratories, suggest that PGF2 alpha and PGB2 induce contraction of the rat aorta, at least in part, by activating PKC. However, PG-induced contractions did not require enhanced phosphoinositide hydrolysis and apparently did not involve the mobilization of intracellular calcium by inositol 1,4,5 trisphosphate. The data thus suggest that PGs activate PKC via a novel mechanism. PMID- 2545868 TI - Defective modulation of noradrenergic neurotransmission by endogenous prostaglandins in aging spontaneously hypertensive rats. AB - Sympathetic nerve stimulation causes a greater vascular response in spontaneously hypertensive rats (SHR) compared to Wistar-Kyoto normotensive rats (WKY), i.e., noradrenergic neurotransmission is enhanced in SHR. Prejunctional and/or postjunctional defects in the regulation of noradrenergic neurotransmission by endogenous prostaglandins could contribute to the increased responsiveness to sympathetic nerve stimulation in SHR. This hypothesis was tested by comparing the effects in SHR vs. WKY of inhibition of cyclooxygenase on vascular responses to periarterial nerve stimulation (PNS), norepinephrine (NE) and angiotensin II (ang II) in the in situ blood perfused rat mesentery. The cyclooxygenase inhibitor, indomethacin, potentiated vascular responses to PNS and NE similarly in 16-week old SHR vs. age-matched WKY. However, in this age group, indomethacin enhanced responses to ang II more in SHR compared with WKY. To determine whether chronic exposure of the vasculature to high blood pressure might alter the physiological significance of prostaglandin-mediated regulation of noradrenergic neurotransmission in vivo, additional studies were conducted in SHR and WKY that were 25 weeks old. In this age group, neither indomethacin nor ibuprofen, an alternative cyclooxygenase inhibitor, significantly potentiated responses to either PNS or NE in SHR, whereas in WKY both indomethacin and ibuprofen potentiated responses to PNS and NE. Also, in these older animals, indomethacin and ibuprofen enhanced responses to ang II equally in SHR vs. WKY. These findings indicate that in aging SHR prostaglandin-mediated regulation of vascular responses to sympathetic nerve stimulation becomes defective. This defect may contribute to the worsening of high blood pressure with age and may be involved in some of the vascular pathology associated with hypertension. PMID- 2545869 TI - Oral hairy leukoplakia is not a specific sign of HIV-infection but related to immunosuppression in general. AB - Oral hairy leukoplakia (HL) has been regarded as an early sign of HIV infection, and its clinical importance related to the poor outcome of the patients has been emphasized. Initially, HL was observed exclusively among male homosexuals, but subsequently demonstrated in all risk groups of HIV infection. The patient described in this article suggests that oral HL is not specific for HIV infection per se, but may be associated with immunosuppression also due to other causes. We describe an HIV-seronegative, heterosexual man suffering from an acute myeloblastic leukemia, who developed clinically and histologically typical HL while on cytostatics. Biopsy showed areas with characteristic ballooning cells, and hyphae of yeasts were demonstrated with PAS-stain. Using the in situ hybridization technique, Epstein-Barr virus (EBV) DNA with high copy numbers was disclosed in the superficial and intermediate cells, whereas human papillomavirus (HPV) DNA (types 6, 11, 16, 18) was not present. PMID- 2545870 TI - Oral hairy leukoplakia in an HIV-negative renal transplant recipient. AB - Oral hairy leukoplakia (HL) has been seen exclusively in those infected with HIV or at risk for AIDS. This case report describes an example of HL seen in a renal transplant recipient who was negative for HIV on serology and culture. The diagnosis of HL was confirmed using in situ hybridization for EBV DNA. PMID- 2545871 TI - Oral manifestations of HIV infection in 75 Dutch patients. AB - Seventy-five consecutive HIV-infected patients, including 33 AIDS cases were examined. One or more oral mucosal lesions were observed in 57 (76%); candidiasis was the most common finding (52%). Others included hairy leukoplakia (16%), periodontal disease (16%) and Kaposi's sarcoma (4%). Diversity of study design and methods allowed no reliable comparison with other reports. PMID- 2545872 TI - Morphometric study of pseudoepitheliomatous hyperplasia in granular cell tumors of the tongue. AB - Several reports have mentioned the possibility of misdiagnosing pseudoepitheliomatous hyperplasia (PEH) of the overlying mucosa of a granular cell tumor as a squamous cell carcinoma (SCC). Because of this, morphometry was applied to five granular cell tumors with PEH and five well-differentiated SCC of the tongue. In addition, ten normal tongues have been examined. The mean area, the mean perimeter and the mean diameter of the 50 largest squamous epithelial nuclei in 50 fields were found to be significantly larger in squamous cell carcinomas than in granular cell tumors and normal tongues. The shape factor of the nucleus and the mitotic activity appeared to be of no significant value in this respect. PMID- 2545873 TI - A mathematical model on the analysis of a one-ligand, one-binding group system when an unknown amount of endogenous ligand is present. AB - In the study of ligand-receptor interaction, binding studies using radiolabelled ligands are usually conducted and results analyzed according to the method originally described by Scatchard. When significant levels of endogenous ligands are present, assumption of a linear relationship between the bound-to-free ratio and the bound ligand may be erroneous and chemical methods have been described to resolve this problem. We present a mathematical approach to the system, using a method based on multiple regression of binding data. A three-dimensional graphic representation of the binding data analogous to the original Scatchard plot is proposed. When the endogenous and the exogenous ligands are different, the linear regression model is applicable only if their affinity constants are known. Non linear curve-fitting computer programs should be used to resolve this problem when these constants are unknown. PMID- 2545874 TI - Hormonal regulation of peripheral benzodiazepine binding sites in female rat adrenal gland and kidney. AB - The effect of hypophysectomy and hormonal replacement on the density of peripheral benzodiazepine binding sites (PBS) in rat adrenal gland and kidney was studied. In the adrenal gland, hypophysectomy caused a significant decrease of 3 fold in PBS density. In the kidney, in contrast, hypophysectomy did not affect PBS density. In the adrenal gland, adrenocorticotropic hormone (ACTH) administered to hypophysectomized rats caused a significant increase of more than 5-fold in PBS density compared to untreated hypophysectomized rats, and of more than 1.6-fold compared to intact rats. In contrast, the hormones pregnant mare serum gonadotropin (PMSG), diethylstilbestrol (DES), and hydrocortisone (HC), administered to hypophysectomized rats, failed to restore PBS density in the adrenal gland. In the kidney, HC administered to hypophysectomized rats caused an increase of 1.4-fold in PBS density compared to untreated hypophysectomized and intact rats. In contrast, the hormones ACTH, PMSG, and DES, administered to hypophysectomized rats, did not affect PBS density in the kidney. None of the hormones tested altered the equilibrium dissociation constant of PBS in either the adrenal gland or the kidney. These findings indicate that PBS density in rat adrenal gland and kidney is hormonally modulated. PMID- 2545875 TI - Effects of sulfhydryl reagents on [3H] inositol trisphosphate binding to dog cerebellar membranes. AB - Homogenates from dog cerebellum were fractionated using sucrose gradient centrifugation. The [3H]inositol 1,4,5-trisphosphate binding and the glucose 6 phosphatase activities were found to co-purify. The binding was saturable and had high affinity (Bmax = 44 pmol/mg protein, Kd = 116 nM). Selective chemical modification was used to examine amino acid residues of the microsomal receptor that might be critical for the binding of inositol trisphophate. Sulfhydryl reagents, p-chloromercuricphenyl sulfonic acid. eosin 5-maleimide, N-ethyl maleimide and fluorescein 5-maleimide were found to be highly potent inhibitors of the binding with half-maximal inhibition occurring at about 20 microM, 70 microM, 1 mM, and 0.1 mM, respectively. The inhibition was specific since the presence of 10 microM of inositol trisphosphate during the reaction completely protected against the inhibition by these reagents. These results suggest that sulfhydryl group is essential for inositol trisphosphate binding to its receptor. PMID- 2545876 TI - Demonstration of type I insulin-like growth factor receptors on human platelets. AB - Human platelets, freshly isolated from healthy human adults, express receptors for insulin-like growth factor I. The IC50 for displacement of 125I-IGF-I binding by unlabeled IGF-I was 0.2 nM, by IGF-II 32 nM by insulin 160 nM. Scatchard analysis of IGF-I binding demonstrates dissociation constants of 0.14 +/- 0.08 nM for high affinity binding site and 54 +/- 18 nM for low affinity binding site. The presence of the alpha-subunit of type I IGF receptor, as high affinity binding site, was verified by affinity crosslinking of 125I-IGF-I to platelet surface membranes. Under reducing con-conditions a Mr = 135,000 band was preferentially labeled. The complete type I IGF receptor complex, which revealed under nonreducing conditions, has an approximately molecular mass of Mr greater than 400,000. The immunoprecipitation of the 125I-IGF-I cross-linked type I receptor with alpha IR-3 confirmed the results achieved by affinity crosslinking. PMID- 2545877 TI - The effects of "matrigenin" activity from bovine bone on the glycosaminoglycans of bovine articular cartilage in culture. A model for cartilage repair by bone derived factors. AB - A bovine articular cartilage longterm culture system was used to study proteoglycan synthesis by a pulse labeling technique. A brief preincubation of the cartilage slices with bacterial collagenase was found to be an effective method of depleting proteoglycan. The addition of a fraction from bovine bone that contained partially purified "matrigenin" activity to the cultures resulted in the stimulation of incorporation of radioactive precursors into cartilage chondroitin sulfate, suggesting increased proteoglycan synthesis. The stimulatory effect of matrigenin activity was observed earlier and was more sustained if the cartilage slices were preincubated with collagenase. The system appears to be useful for testing the hypothesis that bone derived factors stimulate the repair of damaged cartilage. PMID- 2545878 TI - Lymphomatoid granulomatosis in a 13-month-old infant. AB - Lymphomatoid granulomatosis is an infrequent, progressive, and frequently fatal vasculitis which typically occurs in middle aged males. We describe a 13-month old infant with lymphomatoid granulomatosis who presented with chronic otitis media and "failure to thrive." Although it is well recognized that rheumatic conditions may result in failure to thrive, pediatricians often fail to consider this possibility and the proper diagnosis and treatment are correspondingly delayed. Lymphomatoid granulomatosis occurs with increased frequency in immunocompromised patients. In our case both hypogammaglobulinemia and persistent evidence of Epstein-Barr virus infection suggest that the infant was immunocompromised. Whether these factors predisposed this infant to lymphomatoid granulomatosis is uncertain. No patient with the onset of lymphomatoid granulomatosis prior to 7 years of age has been reported. PMID- 2545879 TI - Effects of extracellular pH on neutrophil superoxide anion production, and chemiluminescence augmented with luminol, lucigenin or DMNH. AB - When neutrophils were stimulated with f-met-leu-phe (fMLP) or phorbol myristate acetate (PMA) chemiluminescence (CL) augmented with luminol, lucigenin or 7 dimethylamino-naphthalene-1,2-dicarbonic acid hydrazide (DMNH) showed a strong dependence on the pH of the medium. CL increased in an exponential fashion when pH was increased from 6.5-9.2. All three luminescent intermediaries showed this pH-dependency but luminol was affected most. Unstimulated CL showed a similar increase with pH. Also neutrophil superoxide anion production (in the cytochrome C reduction assay) increased with pH, but not to the same extent as CL. CL induced by xanthine-oxidase plus hypoxanthine increased likewise with pH, and that increase was more pronounced than what was observed from the same reaction in the cytochrome C assay. The lucigenin-amplified neutrophil CL showed the best correlation to the superoxide production from neutrophils. Thus, several effects underlie the variation of CL with pH but the pH dependency of the luminescent intermediaries contribute substantially, which must be considered. PMID- 2545880 TI - Single chloride channels in colon mucosa and isolated colonic enterocytes of the rat. AB - Chloride channels from rat colonic enterocytes were studied using the patch-clamp technique. After removal of mucus, inside-out patches were excised from the apical membrane of intact epithelium located at the luminal surface. They contained spontaneously switching Cl- channels with a conductance of 35-40 pS. The channels were blocked reversibly by anthracene-9-carboxylic acid (1mM). In excised patches from single enterocytes, isolated by calcium removal, the Cl- channels were studied in more detail. The I-V relation was linear between +/- 80 mV. The selectivity was I- greater than Br- greater than Cl- = NO3- greater than F- = HCO3-. Thirty pS Cl- channels were also found on the basolateral membrane of crypts isolated by brief calcium removal. The I-V curve of these Cl- channels was also linear. The results provide direct evidence for the existence of Cl- channels in the apical membrane of surface cells in colonic mucosa. The properties of these channels are similar to those previously observed when incorporating membrane vesicles into planar lipid bilayers. Both results support the validity of the theoretical models describing intestinal secretion. PMID- 2545881 TI - Conductance states activated by glycine and GABA in rat cultured spinal neurones. AB - The conductance properties of single Cl- channels activated by glycine and gamma aminobutyric acid (GABA) were examined in rat spinal cord neurones grown in cell culture. The majority (85%) of spinal neurones were sensitive to both glycine and GABA as were most (83%) outside-out patches tested. Glycine and GABA activated multiple conductance state Cl- channels with linear current-voltage properties when the chloride activities of the solutions bathing both sides of the membrane were similar. Glycine activated six distinct conductance states with conductances of 14, 20, 30, 43, 64 and 93 pS, whereas GABA activated five states with conductances of 13, 20, 29, 39 and 71 pS. The 30 and 43 pS states and the 20 and 29 pS states were observed most frequently with glycine and GABA, respectively. As the values of the glycine- and GABA-activated conductance states form a geometric progression when arranged in ascending order, we concluded that the channels do not consist of a cluster of identical pores. Additional conductance states (50 and 100 pS) were activated by glycine occasionally. The similarity between the conductances of the states activated by the two transmitters is consistent with the proposal that they both activate the same type of Cl- channel. PMID- 2545882 TI - Microelectrode characterization of the basolateral membrane of rabbit S3 proximal tubule. AB - The purpose of this study was to characterize the basolateral membrane of the S3 segment of the rabbit proximal tubule using conventional and ion-selective microelectrodes. When compared with results from S1 and S2 segments, S3 cells under control conditions have a more negative basolateral membrane potential (Vbl = -69 mV), a higher relative potassium conductance (tK = 0.6), lower intracellular Na+ activity (ANa = 18.4 mM), and higher intracellular K+ activity (AK = 67.8 mM). No evidence for a conductive sodium-dependent or sodium independent HCO3- pathway could be demonstrated. The basolateral Na-K pump is inhibited by 10(-4) M ouabain and bath perfusion with a potassium-free (0-K) solution. 0-K perfusion results in ANa = 64.8 mM, AK = 18.5 mM, and Vbl = -28 mV. Basolateral potassium channels are blocked by barium and by acidification of the bathing medium. The relative K+ conductance, as evaluated by increasing bath K+ to 17 mM, is dependent upon the resting Vbl in both S2 and S3 cells. In summary, the basolateral membrane of S3 cells contains a pump-leak system with similar properties to S1 and S2 proximal tubule cells. The absence of conductive bicarbonate pathways results in a hyperpolarized cell and larger Na+ and K+ gradients across the cell borders, which will influence the transport properties and intracellular ion activities in this tubule segment. PMID- 2545883 TI - Digital image analysis of two-dimensional Na,K-ATPase crystals: dissimilarity between pump units. AB - Two-dimensional crystals of purified Na,K-ATPase were induced by treatment with phospholipase-A2 and vanadate. The negatively stained crystals were imaged by electron microscopy and analysed by digital image processing. Two-dimensional averaged projections of the crystals were calculated by the technique of correlation analysis, utilizing SPIDER (System for Processing of Image Data in Electron microscopy and Related fields) image processing software. The calculated dimensions of the unit cell were found to be 13.3 X 4.59 nm with included angle of 98 degrees, comparable to those reported by others. However, the two protomers of the unit cell were found always to be dissimilar in shape and in orientation. All protomers of one side of the dimer ribbon had a triangular outline, and all protomers of the opposing side had a comma shape. This dissimilarity could be explained by two orientations of identical protomers: one orientation for one side of the dimer ribbon, and another orientation for the protomers of the opposing side of the ribbon. An alternative explanation is that the protomers of one side of the dimer ribbon are actually in a conformation different from that of the protomers of the opposing of the ribbon. PMID- 2545884 TI - Effect of reperfusion on the cardiac acetylcholine and norepinephrine contents in rat hearts. AB - The effects of ischemia, reperfusion and hypoxia on the cardiac acetylcholine, choline, norepinephrine and cyclic AMP contents were investigated in isolated, spontaneously beating rat hearts perfused under constant pressure (100 cm H2O) with Krebs-Henseleit solution gassed with 95% O2-5% CO2. Acetylcholine, choline and norepinephrine were determined by high performance liquid chromatography with electrochemical detection. Cyclic AMP was determined by radioimmunoassay. One min reperfusion following 15 min ischemia (termination of perfusion) caused a significant decrease in both cardiac acetylcholine (P less than 0.05) and norepinephrine (P less than 0.01) contents, but had no significant effect on the cardiac norepinephrine/acetylcholine content ratio, or choline or cyclic AMP content. By contrast, 16 min ischemia did not significantly affect the cardiac acetylcholine, norepinephrine, choline or cyclic AMP content. Also, 16 min hypoxia (perfusion with Krebs Henseleit solution gassed with 95% N2 5% CO2) decreased the cardiac norepinephrine content significantly (P less than 0.01) and norepinephrine/acetylcholine content ratio slightly but not significantly. However, hypoxia had no significant effect on the cardiac acetylcholine, choline or cyclic AMP content. Pre-treatment with 10 microns atropine sulfate prevented the decrease in the cardiac acetylcholine content caused by reperfusion but caused a significant depletion in the cardiac norepinephrine content in the control (P less than 0.01) and ischemia (P less than 0.05) groups and a significant decrease in the norepinephrine/acetylcholine content ratio in all three groups (all, P less than 0.05). Extending the reperfusion period to 5 and 10 min following 15 min ischemia also caused a significant decrease in both cardiac acetylcholine and norepinephrine contents compared with the control groups. However, no significant difference in these contents was found between 1 min reperfusion group and 5 or 10 min reperfusion group. Twenty or 25 min ischemia alone did not significantly affect these contents. These findings suggest that reperfusion disturbs both the sympathetic and parasympathetic nervous systems in the heart and that pre-treatment with atropine adversely affects the balance of the autonomic nervous system. PMID- 2545885 TI - Phosphatidylcholine biosynthesis in myocardial ischaemia. AB - In this study the effect of myocardial ischaemia was evaluated on two aspects of phospholipid metabolism: (i) the de novo synthesis of myocardial phospholipids, as indicated by the incorporation of (methyl-3H) choline and (ii) the incorporation of radiolabelled long chain fatty acids into tissue phospholipids. Two models of ischaemia were used namely normothermic ischaemic arrest and hypoxic, low-flow perfusion of the isolated rat heart. The results showed that within 10 min, hypoxic low-flow perfusion significantly inhibited the incorporation rate of (methyl-3H) choline into tissue phospholipids. Since the tissue choline content remained unaltered under these conditions, the results suggested that the de novo synthesis of phosphatidylcholine is very susceptible to ischaemic damage. Inhibition of (methyl-3H) choline incorporation into tissue phospholipids appeared to be due to both a reduction in choline uptake and specific inhibition of the CDP pathway. Perfusion with glucose (10 mM) as substrate completely abolished the ischaemia-induced reduction in (methyl-3H) choline incorporation, indicating that glycolytically produced ATP played an important role in phosphatidylcholine biosynthesis. In contrast to these results, myocardial ischaemia stimulated the incorporation of long-chain saturated and unsaturated fatty acids into tissue phospholipids. In summary, the results obtained showed that myocardial ischaemia profoundly affected phospholipid metabolism which, in turn, might contribute to membrane damage. PMID- 2545886 TI - Interaction of 1,4 dihydropyridine calcium channel antagonists with biological membranes: lipid bilayer partitioning could occur before drug binding to receptors. AB - The binding of dihydropyridine calcium channel agonists and antagonists to receptors in cardiac sarcolemmal membranes is a complex reaction that may involve an interaction with the lipid bilayer matrix of the sarcolemma. Membrane/buffer partition coefficients (lambda) for three dihydropyridine calcium channel antagonists were measured directly in the sarcolemma and sarcoplasmic reticulum membranes and found to be in the range of 5,000 to 150,000. These drugs interact primarily with the membrane bilayer component of these membranes but may also bind to non-receptor proteins. The intrinsic forward rate constants for dihydropyridine binding to sarcolemmal calcium channel receptors were apparently not strongly dependent on their membrane partition coefficients. For example, nimodipine (lambda = 6300) had a forward rate constant of 6.8 +/- 0.6 x 10(6)/M/S, whereas the forward rate constant for Bay P 8857 (lambda = 149,000) was 1.4 +/- 0.8 x 10(7)/M/S. Model calculations for this binding reaction demonstrated that since these drugs are highly lipid soluble, the dependence of these rates on lipid solubility would probably not be reflected in the experimental forward rate constants. In addition, the intrinsic forward rate constant for nimodipine binding to sarcolemmal calcium channel receptors was found not to be linearly dependent on the viscosity of the buffer medium over a five-fold range. The rate of nonspecific (non-receptor protein) drug binding to highly purified sarcoplasmic reticulum membranes essentially devoid of specific receptors for these drugs appears to be extremely fast, at least 10(3) times faster than specific drug binding to the receptor in the sarcolemma. Thus, it appears that partitioning into the lipid bilayer matrix of the sarcolemma could be a general property of dihydropyridine calcium channel antagonists and may be a prerequisite for their binding to sarcolemmal membrane receptors. PMID- 2545887 TI - The specific binding of [3H]EO-122, a radiolabeled class I antiarrhythmic drug to rat heart membranes. AB - [3H]EO-122, a radiolabeled class I antiarrhythmic drug, has been used to characterize a new specific binding system to rat heart membranes. The binding is saturable and competitive with unlabeled EO-122 and other antiarrhythmic drugs. In this system, [3H]EO-122 binds to two sites. Site A with an apparent Kd of 33.5 +/- 1.5 nM, Bmax of 1.05 +/- 0.15 pmol/mg protein and Hill coefficient nH = 4. Site B with an apparent Kd of 233 +/- 25 nM, Bmax equals 5.7 +/- 0.61 pmol/mg proteins and nH = 6. The binding to site B indicates that this site is pharmacologically relevant to known class IA antiarrhythmic drugs such as quinidine and procainamide. Lidocaine (class IB) does not interact with this site. Interpretation of the high Hill coefficient suggests that the binding of an antiarrhythmic drug to its pharmacologically relevant binding site exposes additional binding sites and/or modulates the affinity of adjacent binding sites. PMID- 2545888 TI - Characterization of avian angiotensin II cardiac receptors: coupling to mechanical activity and phosphoinositide metabolism. AB - We have characterized the avian angiotensin II (AII) cardiac receptor and provide data that this receptor couples to both mechanical activity and phospholipid metabolism in the avian heart. In 10-day-old chicks, 125I-AII bound to a high affinity site (Kd = 10 nmol) and a low affinity site (Kd = 79.4 nmol) with estimated binding capacities of 531 and 1330 fmol/mg protein, respectively. The 125I-AII binding was rapid, saturable, reversible and modulated by divalent cations and guanine nucleotides. The potency order for the competitive binding of angiotensin I and II paralleled that observed for in vitro contractile force development in bioassays utilizing left atrial tissue. In avian heart, AII also produced a dose-dependent accumulation of inositol-1-phosphate which was inhibited by the angiotensin antagonist [Sar1, Ile8]AII. The data demonstrate specific avian AII cardiac receptors and provide the first evidence that these receptors couple to both mechanical activity and phosphoinositide metabolism in avian heart. PMID- 2545890 TI - Heat shocks do not mobilize mobile elements in genomes of Drosophila melanogaster inbred lines. AB - Males of three inbred lines of Drosophila melanogaster were heat-shocked 90 min at 37 degrees C. The progenies from treated and untreated males mated with untreated females of the same line were checked for their chromosomal insertion patterns of various mobile elements by either in situ hybridization or Southern blots. No modification in the pattern of insertion of the elements studied was observed after heat treatment. Hence, heating males of our inbred lines did not mobilize mobile elements, contrary to recent reports on other lines of Drosophila melanogaster. PMID- 2545891 TI - Vegetable consumption and lung cancer risk: a population-based case-control study in Hawaii. AB - We conducted a population-based study of diet and lung cancer among the multiethnic population of Hawaii in 1983-1985. We completed interviews for 230 men and 102 women with lung cancer and 597 men and 268 women controls, frequency matched to the patients by age and sex. A quantitative dietary history assessed the usual intake of foods rich in vitamins A and C and carotenoids. A clear dose dependent negative association was demonstrated between dietary beta-carotene and lung cancer risk in both sexes. After adjusting for smoking and other covariates, the men in the lowest quartile of beta-carotene intake had an odds ratio of 1.9 (95% confidence interval, 1.1-3.2) compared to those in the highest quartile of intake. The corresponding odds ratio for women was 2.7 (95% confidence interval, 1.2-6.1). No clear association was found for retinol, vitamin C, folic acid, iron, dietary fiber, or fruits. All vegetables, dark green vegetables, cruciferous vegetables, and tomatoes showed stronger inverse associations with risk than beta-carotene. This observation suggests that other constituents of vegetables, such as lutein, lycopene, and indoles, and others, may also protect against lung cancer in humans. PMID- 2545889 TI - Platyhelminth mitochondrial DNA: evidence for early evolutionary origin of a tRNA(serAGN) that contains a dihydrouridine arm replacement loop, and of serine specifying AGA and AGG codons. AB - The nucleotide sequence of a segment of the mitochondrial DNA (mtDNA) molecule of the liver fluke Fasciola hepatica (phylum Platyhelminthes, class Trematoda) has been determined, within which have been identified the genes for tRNA(ala), tRNA(asp), respiratory chain NADH dehydrogenase subunit I (ND1), tRNA(asn), tRNA(pro), tRNA(ile), tRNA(lys), ND3, tRNA(serAGN), tRNA(trp), and cytochrome c oxidase subunit I (COI). The 11 genes are arranged in the order given and are all transcribed from the same strand of the molecule. The overall order of the F. hepatica mitochondrial genes differs from what is found in other metazoan mtDNAs. All of the sequenced tRNA genes except the one for tRNA(serAGN) can be folded into a secondary structure with four arms resembling most other metazoan mitochondrial tRNAs, rather than the tRNAs that contain a T psi C arm replacement loop, found in nematode mtDNAs. The F. hepatica mitochondrial tRNA(serAGN) gene contains a dihydrouridine arm replacement loop, as is the case in all other metazoan mtDNAs examined to date. AGA and AGG are found in the F. hepatica mitochondrial protein genes and both codons appear to specify serine. These findings concerning F. hepatica mtDNA indicate that both a dihydrouridine arm replacement loop-containing tRNA(serAGN) gene and the use of AGA and AGG codons to specify serine must first have occurred very early in, or before, the evolution of metazoa. PMID- 2545892 TI - mRNA expression of transforming growth factor alpha in human breast carcinomas and its activity in effusions of breast cancer patients. AB - Transforming growth factor alpha (TGF alpha) mRNA expression was measured by Northern blot analysis in 18 human, primary, infiltrating, ductal breast carcinomas. Expression of a 4.8-kilobase TGF alpha mRNA transcript was detected in nine of 18 tumors. No evidence was observed of any gross amplifications or major rearrangements of the TGF alpha gene in the breast carcinoma specimens. Biologically active and immunoreactive TGF alpha was measured in the pleural effusions or in the ascitic fluids from 37 noncancer and 63 cancer patients. The TGF alpha activity detected ranged from 0.2 to 26 ng/mL in most effusions from both groups. However, 29 of 63 (46%) of the effusions from cancer patients exhibited TGF alpha levels that were 6 ng/mL or higher, whereas only seven of 37 (19%) of those from noncancer patients exceeded this level (P less than .03). In particular, effusions obtained from breast cancer patients showed a significantly higher level of TGF alpha, compared with those from noncancer patients (P less than .001). Effusions from 14 cancer patients also contained elevated levels of two tumor-associated antigens, CEA and/or TAG-72, and within this group, nine also had elevated levels of TGF alpha. PMID- 2545893 TI - Needle localization and surgical management of occult breast lesions. AB - Screening mammography can locate small breast cancer lesions not detectable on physical examination. In this study, the records of 57 patients undergoing radiographically guided preoperative needle localization were reviewed for the period August 1986 to May 1988. Of the 57 cases, 15.8% were positive for cancer and 84.2% were benign breast lesions. Invasive ductal carcinoma was the pathologic diagnosis in all malignant biopsies, except for one case of carcinoma in situ. All positive lesions had shown as microcalcifications on mammogram. The authors examine the criteria for biopsy and discuss their experience with needle localization of occult breast lesion suspicious of breast cancer. PMID- 2545894 TI - Redundant colon and carcinoma of the right colon. AB - The combination of redundant colon, multiple villose, adenomas, and a colloid adenocarcinoma arising in one villous adenoma in a black man is rare. The authors report such a case. PMID- 2545895 TI - Expression of nerve growth factor receptors by human peripheral blood mononuclear cells. AB - In the rat, nerve growth factor (NGF) has been shown to affect immune reactivity by binding to cell surface receptors on a subpopulation of splenic mononuclear cells. This binding occurs in a specific and saturable fashion to what appear to be low-affinity (type II) NGF receptors (NGFR). Immunofluorescence studies here showed that NGFR are also present on a proportion of human peripheral blood mononuclear cells (PBMC). Equilibrium binding studies demonstrated that the binding of NGF to its receptors on PBMC occurs with a single equilibrium binding constant (mean) of 2.11 X 10(-9) M. The number of receptors per cell was determined to be approximately 6.94 X 10(3) receptors/cell. These results would suggest a role for NGF in the regulation of immune function in man, as well as in animals. PMID- 2545896 TI - Effects of free fatty acids and acyl-coenzyme A on diacylglycerol kinase in rat brain. AB - Our earlier studies have indicated the presence of diacylglycerol kinase activity in rat brain cytosol as well as subcellular membrane fractions (Strosznajder et al.: Neurochemistry International 8(2):213-221, 1986). There is much evidence indicating the release of diacylglycerols due to stimulation of polyphosphoinositide hydrolysis by hormones and receptor agonists. In turn, diacylglycerols have been linked to a second messenger role for activation of protein kinase C. The present study tests the ability of free fatty acids and acyl-coenzyme A (acyl-CoA) to regulate diacylglycerol kinase activity. In a system containing brain cytosol and microsomes, addition of oleic acid (0.5 mM) resulted in large stimulation of diacylglycerol kinase activity as well as some translocation of the enzyme from cytosol to microsomes. On the other hand, oleoyl CoA (0.1 mM), but neither palmitoyl-CoA nor arachidonoyl-CoA, was effective in translocation of the diacylglycerol kinase. Unlike oleic acid, which preferred to associate with membranes, most of the oleoyl-CoA remained in the cytosolic fraction. Since free fatty acids in brain are stringently controlled and are released during ischemic insult, a condition which also elicits the breakdown of polyphosphoinositide to diacylglycerols, results here suggest a plausible mechanism for regulation of diacylglycerol metabolism by free fatty acids and acyl-CoA. PMID- 2545898 TI - Echocardiographic findings of mobile atrial hepatocellular carcinoma. Report of five cases. AB - Hepatoma with cardiac metastasis is difficult to diagnose antemortem. Herein we describe five cases of hepatoma with intracardiac metastasis detected with two dimensional echocardiography (2DE). The clinical presentations include cardiac murmur, syncope, and chest pain. The 2DE demonstrated a right atrial (RA) mass in each case and the presence of tumor echoes in the inferior vena cava connecting with the RA mass in four. In three cases the cardiac tumor was removed, whereas the other two patients agreed only to liver biopsy, which confirmed the diagnosis. This paper emphasizes the practical utility of 2DE for early detection of intracardiac hepatoma and also describes the clinicopathologic correlation of such a disease entity. PMID- 2545897 TI - Na,K-ATPase: comparison of the cellular localization of alpha-subunit mRNA and polypeptide in mouse cerebellum, retina, and kidney. AB - A clone encoding mouse brain Na,K-ATPase alpha-subunit was isolated from a mouse brain lambda gt11 cDNA library by using antisera to mouse and bovine brain alpha subunit. A comparison of the nucleotide sequence of this clone with published sequences of rat brain alpha-subunit isoform clones showed it to be most similar to rat brain alpha 1. An RNA antisense probe prepared from the cDNA insert of the mouse clone detected a single mRNA of approximately 4.5 kb in Northern blots of mouse brain and kidney RNAs. This probe hybridized only to an alpha 1-cDNA insert from rat brain under high stringency conditions on Northern blots. The RNA antisense probe was used for in situ hybridization to sections of mouse kidney, cerebellum, and retina, and the cellular distribution of alpha-subunit mRNA (alpha-mRNA) was compared with that of alpha-subunit polypeptide (alpha-subunit) detected by immunofluorescence in similar sections. In kidney, alpha-mRNA distribution closely paralleled that of the polypeptide with abundant expression in ascending thick limbs and cortical distal tubules and weaker labeling in cortical proximal tubules. The co-distribution of alpha-mRNA and polypeptide in kidney where Na,K-ATPase localization is well established is consistent with the specificity of these probes. In the retina, prominent labeling with both probes was seen in photoreceptor inner segments, inner nuclear layer, and ganglion cell bodies. Plexiform layers and optic fibers expressed abundant alpha-subunit but little mRNA. Light labeling for both was seen in the outer nuclear layer. In cerebellum, alpha-mRNA and alpha-subunit were associated with soma of granule cells, basket cells, and stellate cells. Glomeruli and basket terminals contained abundant alpha-subunit but exhibited little reactivity with the riboprobe. In Purkinje cell bodies, in contrast, the antibody used to identify the cDNA clone did not resolve significant polypeptide in the somal plasmalemma despite abundant somal mRNA expression. Comparison of distribution of the two probes in cerebellum and retina indicates that message accumulation is primarily in cell bodies, while alpha-subunit epitopes may be co-expressed in cell bodies and/or transported to distant sites in cell-specific patterns. PMID- 2545899 TI - Functional characterization of a complex protein-DNA-binding domain located within the human immunodeficiency virus type 1 long terminal repeat leader region. AB - Transcriptional trans activation of the human immunodeficiency virus type 1 (HIV 1) long terminal repeat (LTR) by the viral tat trans activator is mediated by an LTR-specific sequence located immediately 3' to the start of transcription initiation. We have used a range of molecular techniques to examine DNA-protein interactions that occur in the vicinity of this cis-acting sequence. Our results demonstrate the existence of a sequence-specific DNA-protein interaction involving the HIV-1 leader DNA and map this binding event to between -2 and +21 base pairs relative to the HIV-1 LTR transcription start site. Evidence suggesting that this interaction involves three distinct protein-DNA contact sites extending along one side of the DNA helix is presented. Mutation of these sites was found to ablate protein-DNA binding yet was observed to have no effect on either the basal or tat trans-activated level of HIV-1 LTR-specific gene expression. We therefore conclude that this DNA-protein interaction has a function distinct from the regulation of HIV-1 LTR-specific gene expression. PMID- 2545900 TI - Identification of cellular promoters by using a retrovirus promoter trap. AB - A retrovirus vector has been developed that selects for instances in which the provirus integrates in close proximity to cellular promoters. Coding sequences for a selectable marker (histidinol dehydrogenase) were inserted into the 3' long terminal repeat (LTR) of an enhancerless Molony murine leukemia virus. Although 1.8 kilobase pairs in size, the elongated LTR did not appear to interfere with virus replication or integration. Thus, when the virus was passaged, the elongated LTRs efficiently duplicated, placing histidinol dehydrogenase-coding sequences in the 5' LTR just 30 nucleotides from the flanking cellular DNA. Selection for histidinol expression generated cell clones in which histidinol gene sequences in the 5' LTR were invariably expressed on transcripts initiating in nearby cellular sequences. The efficiency of transducing histidinol resistance was 2,500-fold lower than the efficiency of transducing neomycin resistance when the neomycin phosphotransferase gene was located within the retrovirus and expressed from an independent promoter. By tagging transcriptionally active sites, the vector provides a means to identify and isolate promoters active in different cell types. Furthermore, the virus may be useful as an insertional mutagen, since selection for cell populations containing proviruses only in expressed sites is expected to reduce the number of intergrants needed to screen for loss of gene function. PMID- 2545901 TI - A unique enhancer element for the trans activator (p40tax) of human T-cell leukemia virus type I that is distinct from cyclic AMP- and 12-O tetradecanoylphorbol-13-acetate-responsive elements. AB - The trans activator (p40tax) of human T-cell leukemia virus type I (HTLV-I) is a transcriptional factor that activates the long terminal repeat (LTR) of HTLV-I and interleukin-2 receptor alpha. We examined the HTLV-I enhancer responsible for tax-mediated trans activation and identified (A/T)(G/C)(G/C)CNNTGACG(T/A) as a plausible tax-responsive element (TRE). The putative TRE in the LTR was found to be different from the elements required for activation by cycle AMP and 12-O tetradecanoylphorbol-13-acetate, although these elements overlapped each other. The TRE was also different from a binding site of an NF-kappa B-like factor that was identified in the interleukin-2 receptor alpha promoter and human immunodeficiency virus LTR as a TRE. The latter result was further demonstrated by the failure of the NF-kappa B sequence to compete with the TRE of the LTR in a protein-binding assay. These findings indicate that tax function and its cascade can modulate activities of various enhancer sequences, which are probably regulated by distinct DNA-binding factors. PMID- 2545902 TI - Progesterone and glucocorticoid response elements occur in the long control regions of several human papillomaviruses involved in anogenital neoplasia. AB - We have previously identified in the long control region of the genome of human papillomavirus type 16 (HPV-16) a DNA segment which functions as a cell-type specific enhancer as well as mediating glucocorticoid response. It contains multiple transcription-factor-binding sites, including several for nuclear factor I and one for the glucocorticoid receptor, which binds to the partially palindromic sequence TGTACANNNTGTCAT. We report here that this sequence element, when separated from the surrounding transcription-factor-binding sites and placed as an oligonucleotide into a test vector, retains its function as a glucocorticoid response element (GRE) in HeLa cells. In T47D cells, which express the progesterone receptor, the HPV-16 enhancer fragment mediates progesterone responsiveness. A point mutant in this fragment and the response of the oligonucleotide clone to both steroids prove the identity of the progesterone response element (PRE) with the GRE. The antiprogesterone and antiglucocorticoid RU486 interferes with both hormonal responses. In SiHa cells, the HPV-16 GRE mediates an increase in transcripts encoding E6 and E7 proteins, which are involved in transformation by HPV-16. Hormonal regulation is not restricted to HPV-16: DNA segments containing the cell-type-specific enhancers of HPV-11 and HPV-18 also mediate glucocorticoid and progesterone response. We identified sequence elements in the long control regions of HPV-11 and HPV-18 which function as GRE/PREs when tested as oligonucleotides. These findings suggest that GRE/PREs are an integral part of gene expression regulation in genital HPVs. PMID- 2545903 TI - Expression of two related nonstructural proteins of bluetongue virus (BTV) type 10 in insect cells by a recombinant baculovirus: production of polyclonal ascitic fluid and characterization of the gene product in BTV-infected BHK cells. AB - In vitro translation of bluetongue virus (BTV) double-stranded RNA in the rabbit reticulocyte lysate system has shown segment 10 (S10) to code for two related proteins, NS3 and NS3A. The presence of both products in vivo, however, has remained unconfirmed owing to the very low level of synthesis of the S10 gene product(s) in BTV-infected BHK cells. In the present work, a cDNA copy of BTV type 10 (BTV-10) S10 RNA was inserted into Autographa californica nuclear polyhedrosis baculovirus (AcNPV) in lieu of the 5' coding region of the AcNPV polyhedrin gene. Spodoptera frugiperda cells infected with the recombinant baculovirus synthesized two polypeptides, which were shown to represent NS3 and NS3A by Western blot (immunoblot) and peptide map analysis. Antibodies raised to the expressed NS3 by immunization of mice detected both NS3 and NS3A in BTV-10 infected BHK cells but not in purified BTV-10 virus particles. In contrast to in vitro translation of BTV S10 RNA in which NS3 and NS3A are synthesized in equimolar amounts, NS3 was the principle product both in the baculovirus expression system and in vivo in BTV-infected cells. The results indicate the caution which should be exercised when using the rabbit reticulocyte lysate system to predict the pattern of protein synthesis from a gene with alternative start codons. The expressed NS3 and NS3A proteins reacted strongly with sera from sheep infected with homologous and heterologous BTV serotypes, suggesting that the S10 gene products are highly conserved group-specific antigens. PMID- 2545904 TI - Rotavirus-specific protein synthesis is not necessary for recognition of infected cells by virus-specific cytotoxic T lymphocytes. AB - We found that rotavirus-specific protein synthesis was not necessary for recognition by virus-specific cytotoxic T lymphocytes (CTLs). In addition, CTLs lysed rotavirus-infected target cells prior to production of infectious virus. Target cell processing of rotavirus antigens for presentation to CTLs was enhanced by treatment of rotavirus with trypsin prior to infection; trypsin induced cleavage of the viral hemagglutinin (vp4) has previously been found to facilitate rotavirus entry into target cells by direct penetration of virions through the plasma membrane. We conclude that sufficient quantities of exogenous viral proteins may be introduced into the cytoplasm for processing by target cells. The mechanism by which rotavirus proteins are processed for presentation to the target cell surface remains to be determined. PMID- 2545905 TI - Deletion mutants of herpesvirus saimiri define an open reading frame necessary for transformation. AB - Analysis of a 5,549-base-pair sequence at the left end of herpesvirus saimiri unique (L-) DNA revealed two open reading frames and genes for five small nuclear U RNAs (herpesvirus saimiri U RNAs). Replication-competent deletion mutants were constructed in order to assess the importance of these genetic features for transformation by this oncogenic herpesvirus. Although not required for replication, one of the open reading frames was found to be required for immortalization of marmoset T lymphocytes into continuously growing cell lines. The protein predicted by this reading frame (STP; saimiri transformation associated protein) has a highly hydrophobic stretch of 26 amino acids sufficient for a membrane-spanning domain near its carboxy terminus; this domain is immediately preceded by a sequence appropriate for formation of a metal-binding domain (His X2 His X6 Cys X2 Cys, where Xs are other amino acids). One of two poly(A)+ RNAs that could encode STP is bicistronic, while the other has a long 5' untranslated region (approximately 1.5 kilobases). Although some analogies can be drawn between STP and LMP (lymphocyte membrane protein) of Epstein-Barr virus, STP is not related in sequence to LMP. PMID- 2545906 TI - Induction of anti-EBNA-1 protein by 12-O-tetradecanoylphorbol-13-acetate treatment of human lymphoblastoid cells. AB - Binding of the Epstein-Barr virus (EBV) nuclear antigen (EBNA-1) to BamHI-C DNA was studied by affinity column chromatography followed by immunoblotting with human serum specific for EBNA-1. Two species of EBNA-1 (68 and 70 kilodaltons) were identified in nuclear extracts of the EBV-positive Burkitt's lymphoma cell line Raji and not in nuclear extracts of the EBV-negative Burkitt's lymphoma cell line BJAB. Both EBNA-1s bound specifically to the region required for EBV plasmid DNA maintenance (oriP) located in the BamHI-C fragment. Upon treatment with 12-O tetradecanoylphorbol-13-acetate, which activates latent EBV genome in Raji cells, the 68-kilodalton EBNA-1 was uncoupled from binding to EBV oriP. Nuclear extracts from 12-O-tetradecanoylphorbol-13-acetate-treated BJAB cells also uncoupled the binding of both EBNA-1s to oriP. DNA-cellulose column chromatography identified two protein species which competed for and uncoupled the binding of EBNA-1 to oriP. The two cellular competitors we called anti-EBNA-1 proteins had molecular masses of 60 and 40 kilodaltons, respectively. They were not found in nuclear extracts of BJAB cells not activated by 12-O-tetradecanoylphorbol-13-acetate. PMID- 2545907 TI - Early interactions of pseudorabies virus with host cells: functions of glycoprotein gIII. AB - Adsorption of mutants of pseudorabies virus (PrV) lacking glycoprotein gIII is slower and less efficient than is that of wild-type virus (C. Schreurs, T. C. Mettenleiter, F. Zuckermann, N. Snugg, and T. Ben-Porat, J. Virol. 62:2251-2257, 1988). To ascertain the functions of gIII in the early interactions of PrV with its host cells, we compared the effect on wild-type virus and gIII- mutants of antibodies specific for various PrV proteins. Although adsorption of wild-type virus was inhibited by polyvalent antisera against PrV as well as by sera against gIII and gp50 (but not sera against gII), adsorption of the gIII- mutants was not inhibited by any of these antisera. These results suggest that, in contrast to adsorption of wild-type PrV, the initial interactions of the gIII- mutants with their host cells are not mediated by specific viral proteins. Furthermore, competition experiments showed that wild-type Prv and the gIII- mutants do not compete for attachment to the same cellular components. These findings show that the initial attachment of PrV to its host cells can occur by a least two different modes--one mediated by glycoprotein gIII and the other unspecific. gIII mutants not only did not adsorb as readily to cells as did wild-type virus but also did not penetrate cells as rapidly as did wild-type virus after having adsorbed. Antibodies against gIII did not inhibit the penetration of adsorbed virus (wild type or gIII-), whereas antibodies against gII and gp50 did. It is unlikely, therefore, that gIII functions directly in virus penetration. Our results support the premises that efficient adsorption of PrV to host cell components is mediated either directly or indirectly by gIII (or a complex of viral proteins for which the presence of gIII is functionally essential) and that this pathway of adsorption promotes the interactions of other viral membrane proteins with the appropriate cellular proteins, leading to the rapid penetration of the virus into the cells. The slower penetration of the gIII- mutants than of wild-type PrV appears to be related to the slower and less efficient alternative mode of adsorption of PrV that occurs in the absence of glycoprotein gIII. PMID- 2545908 TI - Identification and expression of a human cytomegalovirus early glycoprotein. AB - A human cytomegalovirus early gene which possesses three temporally regulated promoters is located in the large unique component of the viral genome between 0.054 and 0.064 map units (C.-P. Chang, C.L. Malone, and M.F. Stinski, J. Virol. 63:281-290, 1989). This gene contains a major open reading frame (ORF) located 233 bases downstream of the cap site of an early unspliced RNA. The major ORF predicts a polypeptide of 17 kilodaltons (kDa) which contains a glycoproteinlike signal and anchor domains as well as potential N-glycosylation sites. Antisera were prepared against synthetic peptides derived from amino acid sequences within the major ORF. The antisera detected a viral glycoprotein of 48 kDa in infected cells and recognized the in vitro-translated 17-kDa protein early-gene product. The viral glycoprotein, designated gp48, was modified by N-linked glycans and possibly O-linked glycans. The synthesis of gp48 occurred in the absence of viral DNA replication but accumulated to the highest levels at late times after infection. Since gp48 was found in the virion, it is considered an early structural glycoprotein. PMID- 2545909 TI - A major transcriptional regulatory protein (ICP4) of herpes simplex virus type 1 is associated with purified virions. AB - Herpes simplex virus type 1 was purified by density gradient centrifugation, and the virion-associated proteins were resolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis. By Western blot (immunoblot) analysis with an anti-ICP4 monospecific serum, the results indicated that ICP4, one of the five immediate-early proteins of herpes simplex virus type 1, was associated with the purified virions. To define the location of ICP4 within the virion, trypsin digestion experiments were performed. Purified virions were treated with trypsin in the presence or absence of detergent. The virus envelope appeared to protect ICP4 from the trypsin, since virus-associated ICP4 was sensitive to digestion only after detergent treatment. In addition, ICP4 remained associated with the virus particle when the virion-specific glycoproteins were removed after detergent treatment. Finally, ICP4 was not detected in purified preparations of type A and B capsids isolated from the nuclear fraction of virus-infected cells. The above-mentioned data suggest that detectable amounts of ICP4 are present within the tegument region of the virion. PMID- 2545910 TI - Mapping of functional regions of murine retrovirus long terminal repeat enhancers: enhancer domains interact and are not independent in their contributions to enhancer activity. AB - We have used deletion and recombinant long terminal repeat (LTR) mutants to examine enhancer activity differences between LTRs of the nonpathogenic Akv and the thymus lymphomagenic MCF13 murine retroviruses. Deletion mutant analysis revealed that major control regions for MCF13 and Akv LTR enhancer activity were similar but not identical. For both LTRs, major control regions were distinctly different in a murine T-cell and a fibroblast cell line. Recombinant enhancer analysis showed that LTRs could be divided into three regions capable of altering the level of enhancer activity through cooperative or antagonistic interaction. The contribution of each region to enhancer activity was dependent on its context with respect to the other regions. LTR enhancer function in different cell types appears to be the result of the interaction of enhancer modular elements. PMID- 2545911 TI - Properties of a simian virus 40 mutant T antigen substituted in the hydrophobic region: defective ATPase and oligomerization activities and altered phosphorylation accompany an inability to complex with cellular p53. AB - We have analyzed the biochemical properties of a nonviable simian virus 40 (SV40) mutant encoding a large T antigen (T) bearing an amino acid substitution (Pro-584 Leu) in its hydrophobic region. Mutant 5080 has an altered cell type specificity for transformation (transforming mouse C3H10T1/2 but not rat REF52 cells), is defective for viral DNA replication, and encodes a T that is unable to form a complex with the cellular p53 protein (K. Peden, A. Srinivasan, J. Farber, and J. Pipas, Virology 168:13-21, 1989). In this article, we show that 5080-transformed C3H10T1/2 cell lines express an altered T that is synthesized at a significantly higher rate but with a shorter half-life than normal T from wild-type SV40 transformed cells. 5080 T did not oligomerize beyond 5 to 10S in size compared with normal T, which oligomerized predominantly to 14 to 20S species. In addition, the 5080 T complex had significantly decreased ATPase activity and had a 10-fold-lower level of in vivo phosphorylation compared with that of normal T. Two-dimensional phosphopeptide analysis indicated several changes in the specific 32P labeling pattern, with altered phosphorylation occurring at both termini of the mutant protein compared with the wild-type T. Loss of p53 binding is therefore concomitant with changes in ATPase activity, oligomerization, stability, and in vivo phosphorylation of T and can be correlated with defective replication and restricted transformation functions. That so many biochemical changes are associated with a single substitution in the hydrophobic region of T is consistent with its importance in regulating higher-order structural and functional relationships in SV40 T. PMID- 2545912 TI - Protein kinase activity associated with the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10). AB - The large subunit of the herpes simplex virus type 2 (HSV-2) ribonucleotide reductase (RR1) is demonstrated to possess serine/threonine-specific kinase activity. Computer-assisted sequence analysis identified regions within the amino terminus of ICP10 that are homologous to the catalytic domain of known protein kinases (PKs). An in vitro kinase assay confirmed the ability of ICP10, immunoprecipitated from either HSV-2-infected cells or from cells transfected with an ICP10 expression vector, to autophosphorylate and transphosphorylate exogenous substrates in the presence of ATP and Mg2+. The HSV-1 RR1 was shown to be negative for PK activity under these conditions. PK activity was localized to a 57-kDa amino-terminal region within ICP10 that lacked RR activity. PMID- 2545913 TI - Herpes simplex virus alpha protein ICP27 possesses separable positive and negative regulatory activities. AB - The HSV-1 alpha (immediate-early) protein ICP27 expressed in transfected cells can activate the expression of certain HSV-1 promoters as well as inhibit the transactivated expression of others. We constructed a set of plasmids encoding mutant ICP27 molecules truncated at their carboxyl termini and used transfection assays to determine the functional properties of the mutant proteins. A polypeptide containing the amino-terminal 263 amino acid residues of ICP27 retained partial ability to activate gene expression but was unable to inhibit transactivation. Mutant proteins possessing 406 or 504 amino acids of ICP27 were unable to activate gene expression but retained full ability to inhibit transactivation. These results define two separable regulatory activities of ICP27, one positive and one negative, which can modulate gene expression in transfected cells. Immunoblot and immunofluorescence experiments were used to study the immunological reactivities and intracellular localizations of the mutant proteins. All proteins possessing the amino-terminal 263 amino acids of ICP27 reacted with an ICP27-specific monoclonal antibody and were localized to the cell nucleus. The mutant proteins, however, exhibited a number of phenotypes with regard to intranuclear localization. A mutant possessing 504 residues of ICP27 was similar to the wild-type protein in apparently localizing to all regions of the nucleus. A mutant containing 406 residues of ICP27, on the other hand, was mostly excluded from the nucleolar regions, while a 263-residue mutant was localized predominantly in the nucleoli. Thus, some aspect of ICP27 structure or function can dramatically affect its intranuclear distribution. PMID- 2545914 TI - Neutralizing antibodies specific for glycoprotein H of herpes simplex virus permit viral attachment to cells but prevent penetration. AB - Monoclonal antibodies specific for gH of herpes simplex virus were shown previously to neutralize viral infectivity. Results presented here demonstrate that these antibodies (at least three of them) block viral penetration without inhibiting adsorption of virus to cells. Penetration of herpes simplex virus is by fusion of the virion envelope with the plasma membrane of a susceptible cell. Electron microscopy of thin sections of cells exposed to virus revealed that neutralized virus bound to the cell surface but did not fuse with the plasma membrane. Quantitation of virus adsorption by measuring the binding of purified radiolabeled virus to cells revealed that the anti-gH antibodies had little or no effect on adsorption. Monitoring cell and viral protein synthesis after exposure of cells to infectious and neutralized virus gave results consistent with the electron microscopic finding that the anti-gH antibodies blocked viral penetration. On the basis of the results presented here and other information published elsewhere, it is suggested that gH is one of three glycoproteins essential for penetration of herpes simplex virus into cells. PMID- 2545915 TI - Chimeric picornavirus polyproteins demonstrate a common 3C proteinase substrate specificity. AB - Cross-species proteolytic processing was demonstrated by the 3C proteinases of human rhinovirus 14 and coxsackievirus B3 on poliovirus-specific polypeptide precursors. Chimeric picornavirus cDNA genomes were constructed in a T7 transcription vector in which the poliovirus 3C coding region was substituted with the corresponding allele from human rhinovirus 14 or coxsackievirus B3. In vitro translation and processing of the polypeptides encoded by the chimeric genomes demonstrated that the proteolytic processing of poliovirus P2 region (nonstructural) proteins could be functionally substituted by the heterologous proteinases. In contrast, the 3C proteinase activities expressed from the chimeric genomes were incapable of recognizing the poliovirus-specific processing sites within the capsid precursor. Since the amino acid sequences flanking and inclusive of the P2 region cleavage sites of the three viruses are not stringently conserved, these results provide evidence for the existence of common conformational determinants necessary for 3C-mediated processing. PMID- 2545916 TI - Phorbol ester-inducible T-cell-specific expression of variant mouse mammary tumor virus long terminal repeats. AB - Acquired proviruses of mouse mammary tumor virus (MMTV) in T-cell leukemias of male GR mice have rearrangements in the U3 region of their long terminal repeats (LTR). In contrast to the endogenous nonrearranged MMTV proviruses, these mutated copies are highly expressed in leukemic T cells. To investigate whether the sequence alterations in the LTR are responsible for the high expression of rearranged MMTV proviruses, we made constructs in which normal and variant LTRs drive the bacterial reporter gene chloramphenicol acetyltransferase (CAT). Two different rearranged LTRs were used, one containing a 420-base-pair (bp) deletion (L13) and another carrying a 456-bp deletion plus an 82-bp insertion (L42). These constructs were transfected into murine (GRSL) and human (MOLT-4) T-cell lines that either had or had not been treated with phorbol ester (12-O tetradecanoylphorbol-13-acetate [TPA]). In GRSL cells, the L13-LTR-CAT construct showed transcriptional activity that was further enhanced by TPA. In MOLT-4 cells, both variant LTRs were active, but only after stimulation with TPA. In contrast, normal(N)-LTR-CAT constructs were not expressed, irrespective of TPA addition. In XC rat fibrosarcoma cells, neither normal nor variant LTRs gave rise to detectable CAT activity, either in the presence or in the absence of TPA, but dexamethasone strongly stimulated CAT activity driven by N and L42 LTRs. The L13 LTR was considerably less active, probably caused by the deletion of the distal part of the glucocorticoid responsive element. We conclude that the LTR rearrangements generate TPA responsiveness and contribute to T-cell-specific expression of MMTV variants. PMID- 2545917 TI - Adeno-associated virus P5 promoter contains an adenovirus E1A-inducible element and a binding site for the major late transcription factor. AB - Activity of the adeno-associated virus P5 transcriptional control region was found to be induced by adenovirus E1A gene products. A pair of adjacent sequence elements was found to mediate both basal and E1A-induced P5 activity. The first element is a binding site for the major late transcription factor (MLTF), a factor first identified on the basis of its binding to a specific sequence within the adenovirus major late promoter. The second element is a tandemly repeated 10 base-pair sequence whose relationship to previously described binding sites is unclear. Each element individually conferred E1A responsiveness on a heterologous promoter, and deletion analysis demonstrated that each contributed to the level of P5 activity in the presence of E1A products both in transfection- and infection-based assays. Although deletion of the MLTF binding site led to reduced P5 transcriptional activity in the presence of E1A proteins, the deletion generated enhanced P5 basal activity in the absence of the transcriptional activator. The negative effect of the MLTF binding site in the absence of activator and its positive effect in the presence of activator combine to enhance the magnitude of the response by the P5 control region to E1A gene products. PMID- 2545919 TI - Multiple transcription factor binding sites mediate adenovirus E1A transactivation. AB - We studied the response of simple synthetic promoter regions to transactivation by the adenovirus early region 1A (E1A) protein. Binding sites for one or two host cell transcription factors were substituted for the E1B promoter region in reconstructed virus mutants, and the response to E1A transactivation was assayed during the early phase of infection. We found that a single CREB/ATF binding site resulted in a surprisingly strong promoter which responded to E1A. A CREB/ATF binding site placed upstream of the E1B TATA box behaved much like the wild-type E1B promoter, which is composed of a single Sp1 binding site plus a TATA box. A single E2F binding site resulted in an extremely weak promoter which did not respond to E1A, much like a single Sp1 site. Two E2F sites in an inverted orientation with the same spacing as in the adenovirus type 2 E2 early promoter produced a strong, E1A-responsive promoter. Substitution of the E4 TATA box region for the E1B TATA box region produced a promoter about five times stronger than the wild-type E1B promoter in the absence of E1A. However, the E4 TATA box substitution did not respond significantly to E1A transactivation. These results directly demonstrate that many different transcription factor binding sites, including the E1B TATA box, a CREB/ATF binding site, and two E2F sites, can mediate E1A transactivation. Other transcription factor binding sites cannot mediate an E1A response; these other sites include the E4 TATA box, a single Sp1 binding site, and a single E2F binding site. Implications of these findings for the mechanism of E1A transactivation are discussed. PMID- 2545918 TI - Expression and immunogenicity of the extracellular domain of the human immunodeficiency virus type 1 envelope glycoprotein, gp160. AB - The envelope glycoprotein of human immunodeficiency virus type 1 is synthesized as a precursor, gp160, that subsequently is cleaved to yield mature gp120 and gp41. In these studies, the gene encoding gp160 was mutagenized so as direct the synthesis of a truncated protein consisting of the extracellular domains of both gp120 and gp41. The variant protein, termed sgp160, consisted of 458 amino acids of gp120 and 172 amino acids of gp41. To facilitate protein purification, the normal polyglycoprotein processing site between gp120 and gp41 was deleted through the use of site-directed mutagenesis. This allowed for the synthesis of a molecule that could be purified by affinity chromatography, using acid elution, without dissociation of the gp120 polypeptide from the gp41 polypeptide. The conformation of the sgp160 variant appeared to be functionally relevant, as reflected by its ability to bind to CD4 with an affinity comparable to that of the variant rgp120. The structure of the sgp160-containing polypeptide differed from that of rgp120 in that it tended to form high-molecular-weight aggregates that could be dissociated to monomers and dimers in the presence of reducing agents. Antibodies against the sgp160 protein reacted with authentic virus derived gp160, gp120, and gp41; neutralized viral infectivity; and inhibited the binding of rgp120 to CD4. Rabbit antibodies to the sgp160 protein differed from those raised against rgp120 in that they were enriched for populations that blocked CD4 binding but did not prevent human immunodeficiency virus type 1 induced syncytium formation. PMID- 2545920 TI - Rotavirus-specific cytotoxic T lymphocytes appear at the intestinal mucosal surface after rotavirus infection. AB - The gastrointestinal tract is constantly exposed to a variety of potentially invasive bacteria and viruses. The first line of defense of the host against these pathogens is the intestinal mucosal surface, which consists of epithelial cells, intraepithelial lymphocytes (IELs), mucus, and secretory immunoglobulins. Little is known about the function, memory, or trafficking of IELs after intestinal infection. We found that IELs obtained 6 days after oral inoculation of mice with the intestinal pathogen rotavirus (simian strain RRV) lysed rotavirus-infected target cells; cytotoxic T lymphocytes (CTLs) were responsible for rotavirus-specific cytotoxic activity. Rotavirus-specific cytotoxic activity by IELs was (i) eliminated by treatment with Thy 1.2-specific immunoglobulin M plus complement, (ii) restricted by proteins encoded at the major histocompatibility complex, and (iii) absent in mock-infected animals. Oral inoculation of mice with RRV also induced rotavirus-specific CTLs in splenic and intestinal lymphocytes (mesenteric lymph nodes, Peyer's patch). Parenteral inoculation induced rotavirus-specific CTLs in splenic, intestinal (IELs, mesenteric lymph nodes, Peyer's patch), and nonintestinal lymphocytes (inguinal nodes). Therefore, presentation of rotavirus to the intestinal mucosal surface was not necessary to induce IELs with virus-specific cytotoxic activity. At 4 weeks after oral or parenteral inoculation of mice with RRV, rotavirus-specific CTL precursors appeared among splenic, Peyer's patch, inguinal, and mesenteric node lymphocytes, but not among IELs. IELs with rotavirus-specific cytotoxic activity may be generated from precursors at a site other than the intestinal mucosal surface. Part of the response of the host to enteric infection may include surveillance and lysis of virus-infected villus epithelial cells by IELs. PMID- 2545921 TI - Herpes simplex virus type 1 immediate-early protein Vmw110 reactivates latent herpes simplex virus type 2 in an in vitro latency system. AB - Reactivation of latent herpes simplex virus type 2 (HSV-2) by the immediate-early protein Vmw110 was studied by using an in vitro latency system. Adenovirus recombinants that express Vmw110 reactivated latent HSV-2. An HSV-1 mutant possessing a deletion in a carboxy-terminal region of Vmw110 reactivated latent HSV-2, whereas mutant FXE, which has a deletion in the second exon, did not. Therefore, Vmw110 alone is required to reactivate latent HSV-2 in vitro, and the region of Vmw110 defined by the deletion in FXE is important for this process. PMID- 2545922 TI - Mutational analysis of autonomously functioning trans-activating peptides encoded by adenovirus E1A: evidence for two subdomains. AB - We have shown previously that a chemically synthesized adenovirus E1A region 3 peptide of 49 amino acids, protein domain 3 (PD3; residues 140 to 188 of the 289 amino-acid protein), trans activates viral genes in vitro and in vivo. To study structure-function relationships, we synthesized N-terminal deletion and cysteine substitution mutant peptides and tested their activities in a cell microinjection assay. Peptides lacking 1 to 12 N-terminal residues exhibited 5- to 50-fold reduced molar specific activities, whereas those lacking 16 or 18 residues were inactive. Substitution of each of five PD3 cysteine residues with alanine resulted in substantial losses of activity: mutants in the PD3 N-terminal portion showed 40 to 55% of wild-type activity but required a 20-fold-higher concentration than PD3, whereas those in the C-terminal half were as much less active. These peptide mutant studies suggest the existence of two PD3 functional regions: one, localized in the C-terminal 70 to 75% of the molecule, is essential for trans activation; the other, localized in the N-terminal 25 to 30%, can be overridden to a significant extent at high peptide concentrations. PMID- 2545924 TI - Sendai virus infection of mice with protein malnutrition. AB - Sendai virus pneumonia was produced in BALB/c mice fed protein-deficient diets in an effort to understand the severity of viral pneumonia in infants in developing countries. Animals on the deficient diet became clinically malnourished, and some aspects of cellular immunity were altered. In protein-deprived animals, the 50% lethal dose of intranasally administered Sendai virus was over 1,000-fold lower, pulmonary virus titers were higher, the infection was prolonged, and lung infection was established at a lower inoculum than in normal animals. PMID- 2545923 TI - Transcriptional termination between bovine papillomavirus type 1 (BPV-1) early and late polyadenylation sites blocks late transcription in BPV-1-transformed cells. AB - Bovine papillomavirus type 1 (BPV-1) is a small DNA tumor virus which induces fibropapillomas in cattle and transforms rodent cells in culture. Transcripts are derived from a single strand of the circular viral genome, which has multiple promoters and two polyadenylation sites. In the transformed cell, the first (early) polyadenylation site is utilized exclusively and, therefore, only the early region is expressed. Transcription of the late genes, which requires use of the second (late) polyadenylation site, is seen only in the fully differentiated keratinocytes of the fibropapilloma. In this study, nascent RNA chain analysis of BPV-1-transformed C127 cells was used to demonstrate that at least 90% of the RNA polymerases which transcribe past the early polyadenylation site terminate transcription within the late region before reaching the late polyadenylation site. Therefore, transcription termination is at least partially responsible for the absence of late transcription in the BPV-1-transformed cell and is likely to be an important mechanism for regulation of papillomavirus late transcription during keratinocyte differentiation. PMID- 2545925 TI - Neutralizing monoclonal antibodies against three serotypes of porcine rotavirus. AB - Using three serotypes (four strains) of cultivable porcine rotavirus as immunizing antigens, 10 neutralizing monoclonal antibodies were characterized. One VP4-specific monoclonal antibody directed against porcine rotavirus BEN-144 (serotype G4) neutralized human rotavirus strain ST-3 in addition to the homologous porcine virus. All nine VP7-specific monoclonal antibodies were highly specific for viruses of the same serotype as the immunizing rotavirus strain. One exception was the VP7-specific monoclonal antibody C3/1, which neutralized both serotype G3 and G5 rotaviruses. However, this monoclonal antibody did not neutralize the porcine rotavirus AT/76, also of serotype G3, nor mutants of SA-11 virus (serotype G3) which were selected with monoclonal antibody A10/N3 and are known to have mutations affecting the C antigenic region. PMID- 2545928 TI - Multilocular cyst (multilocular cystic nephroma) of the kidney: a report of 2 cases with an unusual mode of presentation. AB - We report 2 cases of multilocular cyst (multilocular cystic nephroma) of the kidney in children who presented with sudden rapid growth of a renal mass. PMID- 2545927 TI - Carcinoma in situ of the testis. PMID- 2545926 TI - Cytopathic effects induced by Epstein-Barr virus replication in epithelial nasopharyngeal carcinoma hybrid cells. AB - NPC-KT cl.S61, a subclone derived from an epithelial-nasopharyngeal carcinoma hybrid cell line (NPC-KT), showed cytopathic changes characteristic of herpesvirus replication, including formation of multinucleated giant cells and inclusion bodies, when Epstein-Barr virus replicative cycle was induced by 5-iodo 2'-deoxyuridine. Acyclovir (an inhibitor of herpesvirus DNA polymerase), Epstein Barr virus-immune human serum, or 2-deoxyglucose (an inhibitor of the glycosylation) interfered with syncytium formation, indicating that a virus specified glycoprotein belonging to the late group is responsible for cell fusion induced by Epstein-Barr virus replication in cl.S61 cells. PMID- 2545929 TI - Bladder neoplasms associated with augmentation cystoplasty: report of 2 cases and literature review. AB - Bladder neoplasms associated with augmentation cystoplasty have been reported to date in 12 patients. We add 2 cases: 1 with invasive transitional cell carcinoma involving the native bladder and bowel segment, and 1 with a poorly differentiated invasive oat cell (small cell) carcinoma confined to the bladder. The predisposing factors and pertinent literature are reviewed. PMID- 2545930 TI - Suppression of leukocyte function and intracellular content of ATP in hyperosmotic condition comparable to the renal medulla. AB - The effects of hyperosmolality induced by high concentrations of NaCl and urea on the functions of human polymorphonuclear leukocytes was examined. Phagocytosis, intracellular killing of bacteria and superoxide production were inhibited under hyperosmotic conditions. The mechanism of the inhibition of superoxide production was different for NaCl and urea. The inhibition by NaCl was indirect and abolished at 4C and the inhibition by urea was immediate, direct and not abolished at 4C. The intracellular ATP content of leukocytes under hyperosmotic conditions induced by NaCl was significantly reduced and this reduction was abolished at 4C. On the other hand, ATP content under such conditions induced with urea was not reduced. These results suggest that the inhibition of leukocyte function by NaCl is due to an exhaustion of energy source caused by an activation of Na-K pump, and the inhibition by urea is direct inactivation of leukocytes and not due to energy exhaustion. PMID- 2545931 TI - Denervation supersensitivity of the rabbit urinary bladder to calcium ion. AB - We investigated in vitro the supersensitivity of denervated rabbit bladder to Ca2+. Seven days after denervation achieved by bilateral sacral rhizotomy, the capacity of the denervated bladder increased by 2.2 times as compared with the control (126.3 to 277.3 ml.), and wet weight also increased 4.3 times (3.6 to 15.3 gm.). After 30 minutes incubation in normal Krebs' solution the denervated bladder strips showed significantly increased amplitude of contractions to the cumulative acetylcholine addition than the control. After incubation in normal Krebs' solution for 30 minutes, muscle strips were washed three times with Ca free Krebs' solution and incubated 30 minutes in this solution. Cumulative Ca2+ replenishment (0.5 to 10 mM) induced dose-dependent contractions of the detrusor muscle strips in Ca2+-free Krebs' solution, where the contractions were significantly stronger in the denervated detrusor muscle than in controls. Pretreatment with 10(-6) M verapamil (Ca-entry blocker) eliminated the enhanced Ca2+-induced contractions of the denervated detrusor strips, while 10(-6) M procaine (Ca-induced Ca releasing blocker) only partially inhibited them. After depolarizing the cell membrane with 40 mM KCl, the Ca2+-induced contractions of the control strips were markedly enhanced but those of the denervated strips were not. These results demonstrate the supersensitivity of the denervated detrusor muscle to Ca2+. We conclude that hyperpermeability of the cell membrane to Ca2+ influx through calcium channels is responsible for supersensitivity of the denervated detrusor muscle to Ca2+. PMID- 2545932 TI - Cellular congenital mesoblastic nephroma: what are the options. AB - Cellular congenital mesoblastic nephroma is a potentially aggressive variant of the usually benign congenital mesoblastic nephroma. Our recent experience with 3 patients with cellular congenital mesoblastic nephroma prompted a critical review of the literature to evaluate risk factors for recurrence and present treatment programs. A total of 38 patients, including our 3 patients, with cellular congenital mesoblastic nephroma were divided into 2 groups: those with recurrent (7) and those with nonrecurrent (31) tumors. A statistical comparison of clinical and pathological data was performed. Of the 7 patients with local recurrence and/or pulmonary metastasis, 3 died. The average time to first recurrence was 5.4 months. Histological differences were not predictive of recurrent disease. Pathologically positive surgical margins (p less than or equal to 0.02) were the only statistically significant variable suggesting recurrent disease. While the presenting age was not predictive of recurrence (p equals 0.27), the relative risk of recurrence doubles in the first 3 months of life and quadruples after 6 months of life. Treatment programs also were evaluated. An infant with cellular congenital mesoblastic nephroma, regardless of age, is cured with surgery alone given clear pathological margins. In 4 of 5 children with recurrent tumors Wilms tumor treatment agents (vincristine and actinomycin D) failed to control local or distant disease. Of these cases 2 subsequently were treated with sarcomatous chemotherapy (vincristine, cyclophosphamide and doxorubicin) and both are in remission. These agents may prove to be more efficacious in the treatment of local or metastatic disease. PMID- 2545933 TI - Wilms tumorlet, nodular renal blastema and multicystic renal dysplasia. AB - We reviewed 60 cases of multicystic renal dysplasia collected during a 10-year period. Differentiated nonproliferative nodular renal blastema occurred in 6.7 per cent of the cases, which is similar to the incidence of nodular renal blastema in kidneys of other infants with congenital obstructive uropathy. A unique case of cystic dysplasia containing nodular renal blastema and Wilms tumorlet indicates the possibility that a persistently proliferative nephroblastomatous lesion may rarely occur. PMID- 2545934 TI - Nodular renal blastema in the multicystic kidney. AB - Of 60 surgically removed multicystic, dysplastic kidneys examined for the presence of nodular renal blastema only 1 (2 per cent) had blastematous elements. Of the 60 patients 35 were from the authors' personal series and they were examined in clinical detail. The results of this study and those in the literature are discussed in relation to the over-all management of the asymptomatic multicystic, dysplastic kidney. PMID- 2545935 TI - From the Food and Drug Administration. PMID- 2545936 TI - Quantitative estimation of compliance of human systemic veins by occlusion plethysmography with radionuclide--methodology and the effect of nitroglycerin. AB - Volume-pressure relationship and compliance of human systemic veins were estimated quantitatively and noninvasively using radionuclide. The effect of nitroglycerin (NTG) on these parameters was examined. Plethysmography with radionuclide (RN) was performed using the occlusion method on the forearm in 56 patients with various cardiac diseases after RN angiocardiography with 99mTc-RBC. The RN counts-venous pressure curve was constructed from (1) the changes in radioactivity from region of interest on the forearm that were considered to reflect the changes in the blood volume of the forearm, and (2) the changes in the pressure of the forearm vein (fv) due to venous occlusion. The specific compliance of the forearm veins (Csp.fv; (1/V).(delta V/delta P] was obtained graphically from this curve at each patient's venous pressure (Pv). Csp.fv was 0.044 +/- 0.012 mmHg-1 in class I (mean +/- SD; n = 13), 0.033 +/- 0.007 mmHg-1 in class II (n = 30), and 0.019 +/- 0.007 mmHg-1 in class III (n = 13), of the previous NYHA classification of work tolerance. There were significant differences in Csp.fv among the three classes. The systemic venous blood volume (Vsv) was determined by subtracting the central blood volume, measured by RN angiocardiography, from total blood volume, measured by the indicator dilution method utilizing 131I-human serum albumin. Systemic venous compliance (Csv) was calculated from Csv = Csp.fv.Vsv. The Csv was 127.2 +/- 24.8 ml.mmHg-1 (mean +/- SD) in class I, 101.1 +/- 24.1 ml.mmHg-1 in class II and 62.2 +/- 28.1 ml.mmHg-1 in class III. There were significant differences in Csv among the three classes. The class I Csv value was calculated to be 127.2 +/- 24.8 ml.mmHg-1 and the Csv/body weight was calculated to be 2.3 +/- 0.7 ml.mmHg-1.kg-1 of body weight, which was very close to the values for Csv reported previously by other investigators in animal experiments. The administration of NTG caused the RN counts - venous pressure curve to become steeper and shift upward. It increased Csv significantly in all cases. PMID- 2545937 TI - Inhibition of adrenergic transmission by methionine- and leucine-enkephalins is impaired in the mesenteric vasculatures of spontaneously hypertensive rats. AB - The present study was carried out to investigate the effects of enkephalins (methionine-enkephalin: Met-Enk, leucine-enkephalin: Leu-Enk) on the adrenergic neurotransmission in hypertension. Perfused mesenteric vasculatures were prepared in spontaneously hypertensive rats (SHR, Okamoto and Aoki strain, 7-10 weeks old) and age-matched Wistar Kyoto rats (WKY), and the effects of these peptides on vascular responsiveness as well as norepinephrine release from the sympathetic nerve endings were examined. Pressor responses to electrical nerve stimulation were inhibited in a dose-dependent manner by Met-Enk and Leu-Enk, and the inhibition was antagonized by naloxone. Norepinephrine release during electrical nerve stimulation was also suppressed by these peptides. In SHR, stimulation evoked pressor responses and norepinephrine release were significantly enhanced compared to those in WKY, while the suppressive magnitudes of the responses by Met-Enk and Leu-Enk were smaller in SHR than in WKY. These results demonstrate that Met-Enk and Leu-Enk affected presynaptic sites of blood vessels and caused a decrease in electrically-stimulated norepinephrine release from the sympathetic nerve endings. The lower reduction in norepinephrine release and vascular responsiveness by Met-Enk and Leu-Enk in SHR suggests an insufficient regulation of the vascular adrenergic neurotransmission by the opioid peptides in this model of hypertension. PMID- 2545938 TI - [Relation of the serosal invasion mode to a recurrence in advanced gastric carcinomas]. AB - Discussed are the attributable factors affecting the type of carcinomatous recurrence seen in 126 patients who had been operated on for their gastric carcinomas from 1979 to 1982 and who later showed a macroscopically positive serosal invasion (S0) and a histologically ss alpha approximately se invasion and required a curative resection. Forty-six percent of the patients with an ss beta approximately se invasion had a peritoneal recurrence, 21% a liver recurrence, and 33% a recurrence of some other type, whereas those with ss alpha had no such recurrence. Peritoneal recurrence tended to increase with the increase in the length of the serosal invasion and its rates were: one third in serosal invasions of less than 3 cm, one third in invasions of 3 approximately 6 cm and another third in invasions of over 6 cm. The smaller ratio of submucosal length to subserosal length, especially when less than 1.0, meant a greater frequency of peritoneal recurrence. The histologic characteristics of carcinomas that developed a peritoneal recurrence were poorly differentiated, contained INF alpha and had weaker cellular cohesion, whereas those that developed a liver metastasis were well differentiated carcinomas, contained INF beta and had a tighter cellular cohesion. PMID- 2545939 TI - [Two autopsy cases associated with a latent superficial carcinoma of esophagus]. AB - To clarify the histogenesis of squamous cell carcinoma of esophagus, 307 esophagus resected from autopsied cases have been thoroughly examined. These specimens were dyed with Lugol solution and entirely blocked to study subserial sections. Among these specimens, two subclinically superficial squamous cell carcinomas were found. First case uncovered was that of a woman who had died of a carcinoma of uterus. Microscopic examination revealed a small carcinoma in situ, located in the cervical portion of the esophagus, though this lesion showed no associated dysplasia. The other case was that of an old man who had died of a massive hemorrhage from a gastric ulcer, associated with carcinomas of the lip, liver, and prostate. The esophageal lesion was an intramucosal carcinoma located in the mid esophagus that was encountered with moderate dysplasia. These examples are not only quite rare as being multiple primary carcinomas but they also suggest two possible types of cancer development of the esophagus: one that progresses from normal mucosa, and the other from dysplastic mucosa. PMID- 2545940 TI - [A case of quartet cancer--a carcinoma of the breast followed by three synchronous cancers (kidney, thyroid and colon)]. AB - A case of a surgically treated primary quartet cancer is reported, along with a review of the Japanese literature. A 68-year-old woman, curatively operated for a right breast cancer 6 years previously, was then diagnosed as having a left renal cell carcinoma, a thyroid cancer, and a sigmoid colon cancer as a result of a subsequent screening examination within one year. These synchronous, triple cancers were resected in a curative operation for each. Histopathologically, each neoplasm was proven to be a primary cancer lesion, and two different types (papillary and trabecular) were found in the thyroid gland. PMID- 2545941 TI - [Translocation (8; 22) (q24; q11) and Epstein-Barr viral genome in a case of Burkitt's type of acute lymphoblastic leukemia (L3)]. AB - A case of Burkitt's type of acute lymphoblastic leukemia (FAB L3) in an 80-year old male patient is reported. At the time of diagnosis, 74% of the bone marrow cells were lymphoblastoid cells, which were large and homogeneous in size, and whose cytoplasm was abundant and intensely basophilic containing many vacuoles. Lambda-light chains were detected as surface immunoglobulin (Ig), but not heavy chains or kappa-light chains. Epstein-Barr (EB) viral genome was detected in the cultured bone marrow cells by spot hybridization method. Chromosomal banding studies of bone marrow cells revealed t(8; 22) (q24; q11) in all the 18 metaphases examined. This translocation brings the Ig lambda-light chain gene on chromosome 22q11 to chromosome 8q24 where the c-myc proto-oncogene is localized, which is thought to be closely associated with the leukemogenesis of this disorder, whereas EB viral genome is observed in African Burkitt's lymphoma. To our best knowledge, however, there have been no reported Japanese patients with L3 ALL in whom t(8; 22) or EB viral genome was observed. PMID- 2545943 TI - [Regulation of blood coagulation by protein C-thrombomodulin system]. PMID- 2545942 TI - [Hereditary spherocytosis first diagnosed upon the development of aplastic crisis; a case report]. AB - We report a Childhood case of hereditary spherocytosis (HS) first diagnosed upon the development of aplastic crisis. A 6-year-old boy presented with fever and anemia. Although there was neither icterus nor splenomegaly at first, mild icterus and splenomegaly gradually developed with improvement of anemia. The diagnosis of HS was made on the basis of the presence of numerous spherocytes on the peripheral smear, increased osmotic fragility and the auto-hemolysis test result. The severe anemia in the early course with a marked decrease in the bone marrow erythroid cells and the absence of icterus and splenomegaly indicate that it was due to aplastic crisis. In the virological study, anti-human parvovirus (HPV) antibody titers were increased: the values of anti-HPV IgM were high and those of anti-HPV IgG were suddenly elevated. We thus considered that this HS case developed aplastic crisis by HPV infection. PMID- 2545944 TI - [Serpin family proteins in blood coagulation and fibrinolysis (AT III, HC II, PCI, PAI-1, C1INA)]. PMID- 2545946 TI - [Development of antithrombotic drugs and its clinical use--related to platelet function]. PMID- 2545945 TI - [Protein cofactors of coagulation and fibrinolytic systems--high molecular weight kininogen, factor V, factor VIII, thrombomodulin, protein S, and HRG]. PMID- 2545947 TI - [The mode of action and clinical applications of phosphodiesterase inhibitors as anti-platelet drugs]. PMID- 2545948 TI - [Molecular endocrinology: present and prospect]. PMID- 2545949 TI - [Early liver cancer--definition, diagnosis and treatment]. PMID- 2545950 TI - [Molecular markers of hemostatic mechanisms--clinical significance of the measurement of plasma thrombomodulin]. PMID- 2545951 TI - [Rapid detection of herpes simplex virus by DNA probe]. PMID- 2545952 TI - [Histopathological study on human papilloma virus (HPV) infection of the uterine cervix]. PMID- 2545953 TI - [Clinical evaluation of PIVKA-II in patients with hepatocellular carcinoma. Detection of PIVKA-II by EIA using anti-PIVKA-II monoclonal antibody]. PMID- 2545954 TI - [Experience and logic of breast conservation therapy]. AB - One hundred patients with an operable breast cancer were treated with intent of breast conservation in six years. Five patients developed contralateral breast cancers synchronously or metachronously. Therefore, a total of 105 breasts were treated. T1 tumors were seen in 55 breasts, T2 tumors in 45, and T3 in 5. Breast conservation were possible for 95 breasts: 52 with T1 tumors,+ 41 with T2, and 2 with T3. No patients have relapsed yet. There is great concern or even fear for breast conservation among surgeons. We discuss about several issues concerning surgical margin, intraductal carcinoma, multicentricity, radiation injuries including carcinogenesis, racial difference of curability, etc., and try to establish the logic of doing or not-doing this treatment which has been proven by randomized clinical controlled trials. Reasons for conservative attitude expressed by the surgeons are irrelevant or illogical. We conclude that it now is the responsibility for the surgeons to prove survival benefit if they would continue to perform mutilating surgery such as Halsted operation. PMID- 2545955 TI - [The usefulness of motion artifact suppression technic (MAST) in the MRI diagnosis of liver tumors]. AB - The usefulness of motion artifact suppression technique (MAST) in the MRI diagnosis of liver tumors was assessed. Spin-echo with MAST was superior in lesion detection and contrast-to-noise ratio to conventional spin-echo. Its usefulness was verified. As the portal vein is delineated frequently as a high signal area, the portal structure up to the third branches was distinctly visualized. It was found helpful for the segmental diagnosis of the liver. However, careful attention is needed in the diagnosis of portal vein thrombosis. PMID- 2545956 TI - [Adult T-cell leukemia associated with cancer]. AB - We observed adult T-cell leukemia (ATL) associated with cancer in four of 46 patients with ATL, including gastric cancer in two, lung cancer in one, and esophageal and gingival cancer in one. The associated cancer was recognized in three of these patients prior to the diagnosis of ATL and, in one, at autopsy. One patient suffered from a skin type of ATL, two a smoldering type and one a chronic type. No information was obtained from our patients presented here as to whether HTLV-1 influenced the appearance of other cancers, or whether the treatment for cancers influenced the appearance of ATL. Further investigations are necessary to clarify the relationship between ATL and associated cancers. PMID- 2545957 TI - [Lysozyme-positive cells and ultrastructural findings in granulomatous and histiocyte-proliferative skin diseases]. AB - Immunohistochemically, the presence of lysozyme (LZ) has been detected by the antibody against human LZ in cytoplasm of cells from granulomatous and histiocyte proliferative skin diseases. To detect LZ in these cells morphologically, I have done electron microscopic observations of the following skin diseases; sarcoidosis, lupus vulgaris, lupus miliaris disseminatus faciei (LMDF), tattoo granuloma, lichen nitidus, foreign body granuloma, granuloma annulare, xanthelasma, xanthoma tuberosum, xanthoma planum, juvenile xanthogranuloma, giant cell tumor of tendon sheath, dermatofibroma, malignant fibrous histiocytoma, dermatofibrosarcoma protuberans, granulation tissue of burn, hypertrophic scar, and histiocytosis X. From both the immunohistochemical and the electron microscopic features it was concluded that a) immunohistochemically LZ-positive cells from lesions of sarcoidosis, lupus vulgaris, LMDF and tattoo granuloma had a number of electron-lucent bodies (ELB) or microvesicles in their cytoplasm, b) lichen nitidus and xanthoma tuberosum had few LZ-positive cells and the ELB were not observed, and c) the other diseases were LZ-negative, and the ELB were also absent. It is suggested that LZ is present in the ELB which are observed electron microscopically. PMID- 2545958 TI - [Two cases of scleroderma associated with vibration syndrome]. AB - Case 1, a 49-year-old male who had been engaged in repair and reclamation of automobile tires, developed symptoms of vibration syndrome (Raynaud's phenomenon, numbness of both hands, tinnitus and impaired hearing) after some 30 years' use of a grinder and impact wrench. Two years thereafter, multiple sclerodermic lesions appeared over the trunk, upper extremities, and thighs; these disappeared about 2 years later. Histologically, hyperplasia and nodular swelling of collagen bundles were present in the dermis. An immunological study showed the serum to be positive for anti-centromere antibody, but no visceral lesions were demonstrable. This case corresponded to generalized morphea. Case 2, a 53-year-old male, developed symptoms of vibration syndrome (Raynaud's phenomenon, numbness of both hands, impaired hearing and arthralgia) after 25 years' use of a jack hammer in a quarry. Thereafter, sclerodermic changes of the forearms, lower legs, face and abdomen occurred with an associated sclerodactyly. Histological examination of involved skin revealed diffuse hyperplasia and homogenization of collagen bundles throughout the entire thickness of the dermis. These findings, together with serum positivity for anti-RNP antibody and dilation of the lower portion of the esophagus, led us to a diagnosis of progressive systemic sclerosis. We inferred that the vibration syndrome in the present cases might be related etiologically to these forms of scleroderma. PMID- 2545959 TI - [Measurement of superoxide dismutase activity in normal skin by electron spin resonance-spin trapping method]. AB - Superoxide dismutase (SOD) activity in 20 skin specimens from healthy individuals measured by the electron spin resonance-spin trapping method was 7.08 U +/- 0.41 U/mg protein (mean +/- SE). This method may be useful in the determination of SOD, since it requires a shorter time (approx. 3 min) and a smaller amount of specimen (approx. 30 mg) than previously reported methods other than the EIA method. PMID- 2545960 TI - Production of human heterophile antibodies. III. In vitro specific stimulation and immortalization of human lymphocytes. AB - The production of primary human specific antibody against an exogenous antigen by in vitro system was achieved in this study. Human lymphocytes were prepared from the lymphocytes of human spleen (SPL), tonsil (TL), and peripheral blood (PBL). These lymphocytes were stimulated by sheep red blood cells (SRBC) co-cultured with the appropriate number of allogeneic lymphocytes or fractionated T cells. Significant numbers of plaque forming cells (PFC) against SRBC were obtained. A major population of PFC responded to the stimulator RBC and a minor population of PFC responded to both SRBC and bovine red blood cells (BRBC). The culture of allogeneic combination of whole SPL or TL stimulated with SRBC produced PFC, but not that of whole PBL. Reconstitution of equal numbers of allogeneic separated B cells and T cells from PBL was required for a significant response. These specific antibody forming cells (AFC) were immortalized by Epstein Barr virus (EBV) infection and culture supernatant containing antigen-specific antibodies (anti-SRBC: Forssman antibody) were obtained after repeated cloning. PMID- 2545961 TI - Humoral and cell mediated immune responses to porins of Salmonella typhi. AB - Humoral and cell mediated immune responses were studied in control, infected and immunized-infected mice at different time intervals. The levels of antiporin antibodies were found to be higher throughout the study period in immunized infected group in which 87.5% protection was observed by mouse potency test. A significant increase in stimulation index of lymphocyte blast transformation as well as delayed type hypersensitivity response was observed in the same group as compared to infected and control group when porins were used as antigens. Using nonspecific mitogens like PHA and Con A the lymphocyte proliferation was maximum in control group whereas a significant depression was observed in infected mice. It is indicated from this study that porins are excellent antigens interacting efficiently with both arms of the host immune systems which could play a role in providing protection against the disease. PMID- 2545962 TI - [Measurement of total body water with deuterium oxide in renal diseases]. AB - Total body water was measured by means of dilution technic with 99.8% deuterium oxide administration in normal subjects (controls), nephrotic (NS) patients with or without edema, and in longterm hemodialysis (HD) patients. Lean body mass (LBM) was calculated from height, body weight, and waist girth. Body fluid was evaluated according to the TBW/LBM ratio in HD patients. Effect of volume of body fluid on blood pressure was also investigated. Relationship among cardio-thoracic ratio (CTR), standard body weight (SW), and TBW were examined. SWTBW calculated from TBW and LBM, and SWCTR estimated mainly from CTR, blood pressure, were compared. The total body water to body weight ratios (TBW/BW) in controls, NS, and HD patients were 61.2 +/- 1.2%, 71.3 +/- 3.7% (with edema), 60.9 +/- 6.4% (without edema), 64.5 +/- 6.9% (pre-HD), 62.6 +/- 6.6% (post-HD) respectively. The mean TBW/BW of NS patients with edema was significantly greater than those of control, NS patients without edema, and post-HD patients. The TBM/LBM value above 0.78 was suggestive of overhydrated state in HD patients. The mean TBW/LBM in HD patients was 0.757, which was greater than that of controls. Hypotension during HD was seemingly induced by excess ultrafiltration regardless of the value of TBW/LBM. No relationship among CTR, SW, and TBW was observed in HD patients, but the mean TBW/LBM of patients with the CTR exceeding 55% was 0.786, which suggested as overhydrated state. It is useful to determine TBW/BW and TBW/LBM in order to evaluate of volume changes in body fluid of longterm HD patients. PMID- 2545963 TI - [Possible involvement of cyclic GMP (cGMP) in atrial natriuretic factor (ANF) induced natriuresis]. AB - In order to clarify the mechanism of action of ANF on nephron, nephrogenous cGMP (NcGMP) was calculated. And the interrelationship between the changes in NcGMP, urinary sodium excretions or renal hemodynamics were investigated with direct intrarenal infusion of various doses of ANF. A small dose of ANF (0.03 micrograms/kg/min) increased sodium clearance (CNa) significantly without any appreciable changes in NcGMP, creatinine clearance (Ccr) or renal blood flow (RBF). A larger dose of ANF (0.1 or 0.3 micrograms/kg/min) significantly increased CNa, NcGMP, RBF and Ccr. A net increase in CNa (delta CNa) had a positive correlation with that in NcGMP (delta NcGMP), but the regression line did not pass through the zero point. These findings imply that a larger dose of ANF might effect on glomeruli and gene-rate cGMP to induce natriuresis by raising GFR. However natriuresis induces by a small dose of ANF considered to be caused by the other mechanisms, possibly by the direct or indirect effect of ANF on renal tubules. PMID- 2545965 TI - [Application of transbronchial brushing for histological diagnosis of lung cancer]. AB - In order to improve the preoperative diagnostic rate of lung cancer, histological examination was performed by transbronchial brushing in 35 cases, and diagnostic contribution of brushing specimen was compared with that of transbronchial forceps biopsy (TBFB). Histological structures were well preserved in the histology section of brushing specimens, especially in cases of small cell lung cancer. The positive rate of brushing histology was 46%, which was the same as that of TBFB. The total histological diagnostic rate was 66% with simultaneous examination of bronchial brushing and TBFB specimens. Consequently, the use of brushing specimens for histologic evaluation made an additional contribution to the correct positive histology diagnostic rate. Brushing histology is indicated for all types of lung cancer, because in most TBFB-negative cases, biopsies were performed from inappropriate sites of tumors or their materials wer crushed by forceps. It was suggested that concomitant application of brushing specimen for histology examination, contributed to more accurate preoperative histology diagnosis for lung cancer. PMID- 2545964 TI - In vitro characterization of the effects of MCI-154, a novel cardiotonic agent, on cardiac tissues. AB - In vitro cardiac effects of a cardiotonic drug, MCI-154, for which the main action mechanism was proposed to be the enhancement of Ca2+ sensitivity of cardiac contractile proteins, were investigated. MCI-154 (3 x 10(-8)-3 x 10(-4) M) increased the developed tension in isolated ventricular muscles from cats, dogs, guinea pigs and rats and increased that of isolated left atrial muscles of guinea pigs and rats. However, species differences were observed in the responses to MCI-154. The positive inotropic potency of MCI-154 was stronger than those of amrinone and milrinone. In the isolated right atria from guinea pigs and rats, properties of the chronotropic effect of MCI-154 were different from those of amrinone and milrinone. The positive inotropic action of MCI-154 was not affected by phentolamine, propranolol, cimetidine and tetrodotoxin. MCI-154 did not inhibit cardiac Na+,K+-ATPase. MCI-154 moderately stimulated Ca2+-uptake of isolated cardiac sarcoplasmic reticulum (SR), but induced no release of Ca2+ from the SR. These results support the view that the main mechanism for the action of MCI-154 is the enhancement of Ca2+ sensitivity of cardiac contractile proteins. PMID- 2545966 TI - [A case of multiple sclerosing hemangiomas of the lung and a review of the literature in Japan]. AB - A 39-year-old woman complained of an oppressive feeling in her chest and abnormal shadows were detected in the left lower lobe. On a preoperative diagnosis of primary or metastatic lung cancer, left lower lobectomy was performed. There were 3 nodular tumors in the left S6, S10 but no metastasis to regional lymph nodes was found. Histological examination confirmed "so-called" sclerosing hemangioma. She has remained well for 4 years after surgery. A review of the literatures on sclerosing hemangioma in Japan showed that 12 (5%) of 225 cases of sclerosing hemangiomas had multiple tumors. PMID- 2545967 TI - [Analysis of renal cell carcinoma with tumorous legion in the liver]. AB - We presented 3 cases of renal cell carcinoma with hepatic lesion, for which it was difficult to make a diagnosis preoperatively. The hepatic lesion was cavernous hemangioma of the liver, liver metastasis of renal cell carcinoma or hepatocellular carcinoma. To discuss the strategy of treatment for liver metastasis of renal cell carcinoma at the time of nephrectomy, or in the follow up period after nephrectomy, we reviewed the 188 cases of renal cell carcinoma which were nephrectomized from December, 1962 to June, 1988. At the time of nephrectomy, there was only 1 case that had concurrent liver metastasis. In 4 cases, liver metastasis was found at autopsy, and in 15 cases, in the follow-up period after nephrectomy. We analysed these 15 cases and classified them into 2 groups. One was "early metastasis group", i.e., liver metastasis was found within 18 months after nephrectomy, and the other was "late metastasis group", i.e., liver metastasis detected more than 6 years after nephrectomy. In the "early metastasis group", 2 lived 10 months or 57 months, but 5 died within 1 month after the appearance of liver metastasis. In the "late metastasis group", 4 of 7 lived more than 2 years after the appearance of liver metastasis and the median survival was 21 months. In both groups, when liver metastasis was found, there were metastases in multiple organs and the hepatic lesions were multiple.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545968 TI - Lymphocytosis of large granular lymphocytes associated with anemia and neutropenia: proof of monoclonality of the LGL-population, but benign clinical course. AB - A 37-year-old man with severe transfusion-requiring anemia and neutropenia associated with lymphocytosis of large granular lymphocytes (LGLs) has been followed over a period of 3.5 years. Detailed phenotyping of the LGL has been performed, revealing a phenotype of CD3+, CD8+, CD16+, HLA-DR+, and Leu-7+. Southern-blot analysis of the T-cell receptor beta-chain locus detected a gene rearrangement, thus providing proof of monoclonality of the peripheral blood LGLs. PMID- 2545969 TI - [Cryptococcus neoformans meningoencephalitis and multiple infections in AIDS]. AB - A 40-year old homosexual AIDS patient recovering from a Pneumocystis carinii pneumonia developed a Cryptococcus neoformans infection with involvement of the central nervous system (CNS) which could be treated successfully with amphotericin B and flucytosine. After a symptom-free interval of 4 1/2 months, a new acute fatal disease of the CNS did not reveal a cryptococcosis relapse but a necrotizing Toxoplasma encephalitis, a cytomegalovirus infection and striking cultural findings of Staphylococcus aureus in all organs examined. Neither by culture nor by histology Cr. neoformans could be detected in the CNS or in the other organs examined. The temporal course of the Cr. neoformans infection and its specific diagnosis are commented. It is demonstrated that (during or after successful therapy of Pneumocystis carinii pneumonia) a specific cultural examination of specimens from the respiratory tract for Cr. neoformans is needed, in order to recognize a Cr. neoformans infection in its primary stage, i.e. before hematogenous dissemination of Cr. neoformans leading to the secondary stage of the infection. PMID- 2545971 TI - Functional and morphologic expressions of trophoblast. PMID- 2545970 TI - [Effect of low-energy laser radiation on the levels of cyclic nucleotides and lactic acid of the mucous membranes in gastric and duodenal ulcers]. AB - The level of cyclic nucleotides in biological material undergoes changes due to endoscopic treatment of gastric and duodenal ulcers with the help of helium-neon laser radiation which, in addition to dynamics of the clinical picture and gastroduodenoscopic data, makes it possible to diagnose the disease remission and in certain cases to predict the course of peptic ulcer. PMID- 2545973 TI - Alternating methotrexate and dactinomycin in nonmetastatic gestational trophoblastic disease. AB - Alternating 5-day chemotherapy with methotrexate and dactinomycin as primary therapy for nonmetastatic gestational trophoblastic disease was studied in nine patients. The complete response rate was 100% with follow-up of a median of 80 months. Stomatitis was universal but rarely prevented oral alimentation or delayed therapy. Overall, 94% of toxicity was mild or moderate in severity and all toxicity was reversible. This alternating non-cross resistant regimen, reported in a total of 40 patients in the literature, is the only regimen to result in a 100% response rate. This response rate is statistically improved when compared to historical controls receiving methotrexate/folinic acid or pulse dactinomycin. No patients required hysterectomy for disease control. Cooperative prospective phase III studies are needed to determine the efficacy and toxicity of current regimens. PMID- 2545972 TI - Role for T lymphocytes in silica-induced pulmonary inflammation. AB - The role of the immune system in modulating pulmonary damage evoked by exposure to silica crystals has been examined in a mouse model of experimental silicosis. Congenitally T cell-deficient mice (Balb/c nu/nu) were compared with T cell sufficient mice (Balb/c nu/+) in the degree of silica-induced cellular inflammation on days 3, 5, 7, 30, and 60 after an intratracheal instillation of silica. Cellular inflammation was assessed by increases in the total number and type of inflammatory cell in lung lavage fluid and in lung tissue. Inflammation and/or injury were also measured in the lavage fluid by increases in total protein, angiotensin-converting enzyme and plasminogen activator. No significant difference existed between the two types of mice in total lavage cell number or total protein. In nu/+ mice, the neutrophil predominated early, followed by the macrophage as the major inflammatory cell. However, in nu/nu mice, the neutrophil remained the predominant inflammatory cell throughout the two months postinjection. In addition, angiotensin-converting enzyme levels remained elevated in the nu/nu animals above those in the nu/+ mice whereas plasminogen activator was elevated early (before day 7) in both, and decreased comparably over time. These data suggest that T cells directly or indirectly influence the maintenance of a macrophage infiltrate and the termination of a neutrophil response after exposure to silica. However, it appears that neither T cells nor the cells they influence affect the ultimate amount of collagen deposition. PMID- 2545974 TI - Complications of hypogastric artery cisplatin infusions. AB - In an attempt to increase the therapeutic index of Cisplatin (CDDP), 29 continuous 48 hour intra-arterial (IA) infusions of 100-150 mg/m2 CDDP were given to 26 patients with bulky stage T3-T4 bladder cancer. Hypogastric artery catheters were placed distal (n = 42) or proximal (n = 7) to the origin of the superior gluteal artery. Atherosclerosis (n = 5) or aneurysm (n = 2) prevented successful IA catheter placement. Catheter maintenance resulted in no bleeding or thromboembolic episodes. Urethral catheters caused two urinary tract infections. Systemic toxicity was mild with on 4/29 infusions resulting in WBC less than 3,000, 3/29 infusions resulting in creatinine elevation, and 1/29 in peripheral neuropathy. Local effects of the IA CDDP included gluteal pain and ecchymosis (n = 1) and moderately disabling lower extremity neuropathies (n = 3). Systemic side effects of CDDP can be diminished by use of IA route of administration and slow continuous infusion. PMID- 2545975 TI - Ruptured hepatocellular carcinoma evokes risk of implanted metastases. AB - In three patients with hepatocellular carcinoma (HCC), implanted metastases occurred in the peritoneal or pleural cavity after rupture of the tumors; in one patient this was caused by trauma, and in two others this complication became obvious during resection of the tumor. Hepatic resection was successful, and the postoperative status was satisfactory. Implanted metastases were present in the peritoneal or pleural cavity of these three patients 6 years, 10 months, and 6 months after surgery, respectively. We wish to emphasize that a ruptured HCC may lead to implanted metastases. PMID- 2545976 TI - Effects of active compounds isolated from Angelica shikokiana on lipid metabolism in fat cells. AB - It was found that the EtO Ac fraction of the roots of Angelica shikokiana inhibited adrenaline-induced lipolysis in rat fat cells, while having no effect on ACTH-induced lipolysis. On the other hand, the MeOH fraction was found to inhibit ACTH-induced lipolysis but not to inhibit adrenaline-induced lipolysis. The active substances isolated from this root were elucidated to be psoralen (I), bergapten (II), isopteryxin(3'-angeloyloxy-4'-acetoxy-3',4'-dihydroseseli n) (III) and 3'-epoxyangeloyloxy-4'-acetoxy-3',4'-dihydroseselin (IV), respectively. The two furocoumarins, psoralen and bergapten, were found to enhance ACTH- and adrenaline-induced lipolysis in fat cells respectively. In contrast, a coumarin derivative of khellactone type, isopteryxin (III), was found to inhibit the adrenaline-induced lipolysis but with no effect on ACTH-induced lipolysis and insulin-induced lipogenesis from glucose. The other coumarin of khellactone type, 3'(R), 4'(R)-3'-epoxyangeloyloxy-4'-acetoxy-3',4'-dihydroseseline (IV), was found to inhibit the insulin-induced lipogenesis from glucose but it had no effect on adrenaline- and ACTH-induced lipolysis. PMID- 2545978 TI - Rapid shell vial culture and tissue histology compared with serology for the rapid diagnosis of cytomegalovirus infection in liver transplantation. AB - Sixty-six patients who had undergone 78 liver transplantations, with no detectable cytomegalovirus (CMV) infection before transplantation, were studied to assess the value of CMV serology compared with the rapid detection of the virus in shell vial cell cultures or histology for the diagnosis of (1) the first evidence of infection, (2) symptomatic disease, and (3) asymptomatic infection. Of 28 evaluable patients, shell vial cell culture assay and histologic findings provided the first evidence of CMV infection earlier in 22 (79%) or at the same time as the serologic response in 5 (18%). Serologic results yielded the first indication of CMV infection in only 1 of these 28 patients (3%). Of 17 evaluable transplantations in 15 patients in whom symptomatic invasive disease developed, shell vial culture or histology provided a laboratory diagnosis of CMV infection earlier or at the same time as serologic responses in 16 (94%). Similarly, shell vial culture or histology provided evidence of CMV infection in 10 of 11 patients (91%) who had asymptomatic CMV infection and remained asymptomatic. Urine surveillance cultures yielded the first evidence of CMV infection in 14 of 19 patients who initially had an asymptomatic infection, of which 6 became symptomatic. Culturing of the blood with use of the rapid shell vial technique showed that viremia preceded CMV organ involvement in 7 of 10 patients. We recommend culture by the shell vial assay as the most rapid and sensitive method of determining CMV infection in liver transplant patients. PMID- 2545977 TI - Ecotropic virus involvement in spontaneous B-cell lymphomas of CWD/LeAgl mice. AB - The inbred mouse strain CWD/LeAgl, which has a high incidence of spontaneous B cell lymphomas, expresses both ecotropic MuLV and MCF viruses. Studies indicated that the MCF viruses expressed in CWD tumors were characteristic of nononcogenic MCF viruses and that ecotropic MuLV may be the etiological agent in spontaneous B cell lymphomagenesis. Somatically acquired proviruses of approximately the same size were detected in several tumor DNAs suggesting that integration of proviral sequences into specific regions of the mouse genome may be an important step in lymphomagenesis of this strain. PMID- 2545980 TI - [Chronic fatigue syndrome associated with Epstein-Barr virus infection]. AB - Epstein-Barr virus (EBV) infection is ubiquitous and may result in multiple and widely different clinical features; the most common of these is infectious mononucleosis (IM). Recently, a group of patients has been included in the chronic EBV infection syndrome (EBVIS), with a sustained nonspecific syndrome consisting of asthenia, anorexia, low grade fever and changes in mood, associated with a viral infection not necessarily caused by EBV; this has been called chronic fatigue syndrome (CFS). We report a patient who fulfilled the criteria for CFS associated with EBV after an acute, well documented EBV infection. We discuss its etiological and pathophysiological implications, emphasizing the need for extreme caution in the diagnosis of CFS. A merely clinical diagnosis may hide severe mistakes. PMID- 2545981 TI - [Myelodysplastic syndromes and solid neoplasms. Presentation of a case showing an association of refractory anemia and glioblastoma multiforme]. PMID- 2545979 TI - The role of ifosfamide and cyclophosphamide in the multi-modality treatment after surgery for cure for small-cell bronchial carcinomas (SCLC). AB - For the optimisation of the therapy for small cell bronchial carcinomas (SCLC), surgery is used to eliminate the primary tumor and its regional lymph nodes and chemo- and radiotherapy for the general treatment of micrometastasis. After patho histological examination of the operation specimen, randomization for two arms is performed for a standard chemotherapy (CAV) or a sequential chemotherapy using three different drug combinations. Thereafter all disease-free patients receive prophylactic cranial irradiation (PCI). Preliminary evaluations in December 1987, of 112 patients from 19 cooperating departments show that the survival rate projected for 2 yr of 43 patients at stage pT1-3 N0 M0 is 76%, of 43 patients at stage pT1-3 N1 M0 it is 63% and of 26 patients at stage pT1-3 N2 M0 it is 38%. PMID- 2545982 TI - Inhibitory effects of (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU) and related compounds on herpes simplex virus (HSV)-infected cells and HSV thymidine kinase gene-transformed cells. PMID- 2545983 TI - Malignant germ cell tumors of the head and neck in childhood. AB - From 1962 through 1987, four children were diagnosed at our institution with primary germ cell malignancies of the extracranial head and neck regions. Ages of the children ranged from 2 to 44 months. Histologic findings included 2 yolk sac carcinoma (endodermal sinus tumor), 1 malignant teratoma with nephroblastoma (Wilm's tumor), and 1 malignant teratoma with neuroblastoma (primitive neuroectodermal) components. Complete clinical and surgical staging was performed to rule out additional sites of disease. All patients initially underwent either biopsy or, when technically feasible, resection. Three patients received combination chemotherapy and two received irradiation. Three patients died of progressive disease. One patient who had yolk sac carcinoma of the temporomandibular region is alive and free of disease 40 months after therapy. Complete surgical resection is indicated for teratomatous tumors, if technically feasible. The malignant components of these tumors are sensitive to both chemotherapy and irradiation and combined therapy may be beneficial. PMID- 2545984 TI - Further studies on growth characteristics of Epstein-Barr virus (EBV) genome positive and -negative epithelial hybrid cell lines. AB - Differences in tumorigenicity between the Epstein-Barr virus (EBV) genome negative and -positive epithelial A2L/AH hybrid cell lines have been described recently. Further differences in growth characteristics of the two epithelial hybrid cells have since been examined. Differences were found in doubling time and cloning efficiency in agarose of the two cell lines. In addition, the EBV genome-negative epithelial hybrid cell line (cl-654), unlike the EBV genome positive epithelial hybrid cell line (cl-1), failed to grow in the unmanipulated nude mice, but was successfully transplanted in 60Co-irradiated nude mice. The histopathologic findings of tumors induced by the inoculation of cl-654 cells in nude mice showed more differentiated carcinoma than the findings of tumors induced by cl-1 cells. The data suggest that EBV is associated with an alteration of cell differentiation in epithelial cells as well as tumorigenicity in nude mice. PMID- 2545985 TI - Gestational changes of GABA levels and GABA binding in the human uterus. AB - The concentrations of gamma-aminobutyric acid (GABA), the activities of L glutamate decarboxylase and GABA-transaminase, and the nature of the sodium independent binding of GABA were examined in uterine tissue pieces obtained surgically from pregnant and non-pregnant women. GABA concentrations were reduced, while the activity of GABA-transaminase and the specific binding of [3H]GABA significantly increased in specimens from pregnant subjects. These findings suggest some gestation-related functional role for the GABA system in the human uterus. PMID- 2545986 TI - Levels of viral glycoprotein provide a dose-dependent measure of tumor cell inhibition. AB - A chemosensitivity assay utilizing small replicate Mm5mt/c1 C3H mammary tumor cell cultures was developed to determine whether changes in viral antigen expression and release into culture fluids could be utilized as an in vitro measure of chemotherapeutic drug effect. The 52,000 MW viral envelope glycoprotein (gp52) of the mouse mammary tumor virus (MMTV) was measured in culture fluids of control and drug-treated cultures while cell density was simultaneously determined by cell staining and OD 664 mu determination. While extracellular gp52 levels and cell density both progressively increased over 72 hours for control cultures, treatments with doxorubicin resulted in dose dependent declines in both parameters at 24, 48, and 72 hours. Comparison of doxorubicin dosages for 50% reduction (ED50s) in both parameters (0.68uM and 1.1uM) revealed a similar coordinate reduction in both cell density and MMTVgp52. When gp52 levels were further examined as a general measure of effect for a broad spectrum of 6 drugs with differing mechanisms of action, coordinate declines in cell density and MMTVgp52 provided a time and dose-dependent dual measure of effect for each of the drugs tested. Coordinate declines resulted in the same following hierarchy of concentration-dependent drug potency: methotrexate greater than 5-fluorouracil greater than doxorubicin greater than N [phosphonacetyl- L aspartic acid] (PALA) greater than cis-platinum greater than cyclophosphamide. The dual measures of therapeutic effect afforded by this assay argue for its use as an in vitro measure of effect for optimizing drug treatments. PMID- 2545987 TI - Adrenergic receptor homologies in vertebrate and invertebrate species examined by DNA hybridization. AB - The deduced protein sequences of the mammalian adrenergic receptors (ARs) suggest that these proteins have evolved by several ancient gene duplication events. To investigate in what species these events may have occurred DNA fragments encoding the family of adrenergic receptors from human (beta 1AR and alpha 2AR) and hamster (beta 2AR and alpha 1AR) were used to detect homologous sequences in other vertebrates, invertebrates and unicellular organisms by Southern blot hybridization analysis. Sequences homologous to hamster beta 2AR were detected in lower vertebrates, invertebrates and Dictyostelium, but not in yeast or bacteria. Within vertebrates, sequences strongly homologous to human beta 1AR and human platelet alpha 2AR were confined to the higher vertebrates only. In the invertebrates, only Drosophila contained sequences homologous to hamster alpha 1AR. Our results suggest that non-mammalian species may contain receptors homologous to the mammalian adrenergic receptors and that the sequences homologous to human beta 2AR have been the most strongly conserved. PMID- 2545988 TI - Beta-adrenergic enhancement of angiotensin II-stimulated aldosterone secretion. AB - Beta-adrenergic agonists have been shown to stimulate aldosterone secretion. Angiotensin II (AII) is one of the important stimuli of aldosterone secretion; conceivably beta-adrenergic influences affect the stimulatory potential of AII. Using cultured rat adrenal capsules, we found that 10(-7) M epinephrine and 10( 7) M isoproterenol enhanced 10(-7) M AII-stimulated aldosterone production. Propranolol (10(-7) M) completely inhibited the ability of epinephrine to augment the stimulatory actions of AII. In conclusion, beta-adrenergic agonists promote stimulation of aldosterone secretion by AII. PMID- 2545989 TI - Inhibition of serotonin release by bombesin-like peptides in rat hypothalamus in vitro. AB - We investigated the activity of bombesin (BN), neuromedin-C (NM-C) and neuromedin B (NM-B) on serotonin (5-HT) release and reuptake in rat hypothalamus (HYP) in vitro. BN and NM-C but not NM-B (all 1 microM) decreased K+ evoked 3H-5-HT release from superfused HYP slices by 25%. Bacitracin (BCN, 2 micrograms/ml), a nonspecific peptidase inhibitor, reversed the inhibitory effect of BN on K+ evoked 3H-5-HT release. Phosphoramidon (PAN, 10 microM) an endopeptidase 24.11 inhibitor, abolished the inhibitory effect of BN, but not NM-C, on K+ evoked 3H-5 HT release. The peptidyl dipeptidase A inhibitor enalaprilat (ENP, 10 microM), enhanced both BN and NM-C inhibition of 3H-5-HT release. Bestatin (BST, 10 microM) had no effect on BN or NM-C inhibitory activity on 3H-5-HT release. Neither BN, NM-C nor NM-B affected reuptake of 3H-5-HT into HYP synaptosomes alone or in combination with any of the peptidase inhibitors, nor did these peptides alter the ability of fluoxetine to inhibit 3H-5-HT uptake. These data suggest: a) that BN-like peptides may alter neurotransmission in the HYP by acting presynaptically on the 5-HT release mechanism; b) a similarity in the structural requirements for the BN induced inhibition of 5-HT release and BN evoked thermoregulatory disturbances; and c) that peptidases may selectively augment or reduce pharmacologic activity of BN-like peptides upon CNS administration. PMID- 2545990 TI - Effects of streptozotocin-induced diabetes on feeding stimulated by centrally administered opioid agonists. AB - The potencies of several opioid agonists are reduced in diabetic animals and in animals made hyperglycemic via injections of glucose. In this report we examined the effects of streptozotocin-induced diabetes on the feeding responses to centrally administered opioid agonists with differing receptor selectivities. The selective mu receptor agonist Tyr-D-Ala-Gly-(Me)Phe-Gly-ol (DAGO) caused a larger increase in intake in diabetic rats than in controls. In both groups feeding responses were greater on the fourth day of daily injections than on the first day. The delta receptor agonist [D-Ser2,Leu5]-enkephalin-Thr6 (DSLET) stimulated intake in controls but not in diabetics. However, the elevated baseline and large variability in intake of the diabetics in this experiment prevent drawing a conclusion on diabetes-induced changes in the potency of this peptide. No differences between controls and diabetics were apparent in the feeding responses to U50, 488H, a selective kappa receptor agonist. These data suggest that diabetes may differentially affect the classes of opioid receptors or the binding of ligands to these receptors. PMID- 2545992 TI - Cholecystokinin-8 suppressed 3H-etorphine binding to rat brain opiate receptors. AB - Radio receptor assay (RRA) was adopted to analyse the influence of CCK-8 on 3H etorphine binding to opiate receptors in rat brain synaptosomal membranes (P2). In the competition experiment CCK-8 (1pM to 1 microM) suppressed the binding of 3H-etorphine. This effect was completely reversed by proglumide at 1 microM. Rosenthal analysis for saturation revealed two populations of 3H-etorphine binding sites. CCK-8 (1pM to 1 microM) inhibited 3H-etorphine binding to the high affinity sites by an increase in Kd (up to +235%) and decrease in Bmax (up to 80%) without significant changes in the Kd and Bmax of the low affinity sites. This effect of CCK-8 (10nM) was also completely reversed by proglumide at 1 microM. Unsulfated CCK-8 (100pM to 1 microM) produced only a slight increase in Kd of the high affinity sites (+64%) without affecting Bmax. The results suggest that CCK-8 might be capable of suppressing the high affinity opioid binding sites via the activation of CCK receptor. PMID- 2545991 TI - Contribution of 4-hydroxy-2,3-trans-nonenal to the reduction of beta-adrenoceptor function in the heart by oxidative stress. AB - Oxidative stress reduces adenylate cyclase activity and also the maximal response to beta-adrenoceptor stimulation in the rat heart, while beta-adrenoceptor density is not affected or increased. Since free sulfhydryl groups are essential to beta-adrenoceptor function and the sulfhydryl reactive substance 4-hydroxy-2,3 trans-nonenal (HNE) is responsible for part of the effects of oxidative stress, the effect of HNE on beta-adrenoceptor function in field stimulated left atria of the rat was determined. To this end field stimulated atria were incubated with 10 microM, 100 microM and 1 mM HNE for 25 min. After removing the excess of HNE, beta-adrenoceptor function was determined by measuring the positive inotropic response to (-)-isoproterenol. It was found that 10 microM HNE had no effect on beta-adrenoceptor function, whereas 100 microM HNE reduced the maximal effect to (-)-isoproterenol without affecting the pD2 (-log EC50). At these concentrations, HNE had no effect on either beta-adrenoceptor density or on c-AMP production. After 1 mM HNE, the atria stopped contracting. Since the effects of the synthetic thiol inactivator N-ethyl maleimide were similar to those of HNE, it was concluded that the reduction of beta-adrenoceptor function by HNE is probably the result of alkylation of free sulfhydryl groups. Our results indicate that the reduction of adenylate cyclase activity by oxidative stress is not mediated by the production of HNE, however oxidative stress and HNE both reduce the maximal response to beta-adrenoceptor stimulation. PMID- 2545993 TI - Dexamethasone suppresses ACTH release without attenuating pituitary cyclic AMP response to stress in vivo. AB - Dexamethasone, a synthetic glucocorticoid, has been shown to decrease basal and stress-elevated levels of the pituitary hormone ACTH. Glucocorticoids are known to bind to multiple sites within the brain and pituitary and it is not known which site(s) is most important in mediating the observed inhibition of ACTH release. At the level of the corticotroph, there is contradictory data from in vitro studies regarding whether dexamethasone acts proximal or distal to the formation of the cyclic AMP second messenger that has been shown to be involved in CRF-stimulated ACTH release. In the present report, we have examined the effects of dexamethasone pretreatment on stress-induced elevations in pituitary cyclic AMP and the release of ACTH in vivo. Acute stress (15 min of intermittent footshock) elevated levels of pituitary cyclic AMP and plasma ACTH consistent with previous studies. Dexamethasone administration (0.4 mg/kg 24 hr prior to sacrifice plus 0.2 mg/kg 2 hr prior to sacrifice) inhibited stress-induced elevations in plasma ACTH but did not affect pituitary cyclic AMP response to acute stress. These findings suggest that dexamethasone inhibits the release of ACTH via an action distal to the generation of cyclic AMP. PMID- 2545994 TI - Increased serum levels of endopeptidase 24.11 ('enkephalinase') in patients with end-stage renal failure. AB - Endopeptidase 24.11, a widely distributed membrane-bound peptidase is found in low levels in the serum of normal individuals. Although increased levels of the enzyme have been found in sera of patients with sarcoidosis and adult respiratory distress syndrome, the cellular origin of circulating endopeptidase 24.11 remains unknown. As the brush border of the proximal tubular epithelial cells have the highest endopeptidase specific activity, we investigated the possible contribution of the kidney to the release of endopeptidase 24.11 in the systemic circulation. Therefore, we measured serum levels of the enzyme in patients with end-stage renal failure (ESRF) treated by haemodialysis (HD) or continuous ambulatory peritoneal dialysis (CAPD). Increased serum levels of endopeptidase 24.11 were observed both in HD patients (mean +/- SEM: 74.6 +/- 20.9 ng/ml) and in CAPD patients (mean +/- SEM: 45.1 +/- 8.1 ng/ml) as compared to normal individuals (mean +/- SEM: 13.6 +/- 1.4 ng/ml). Endopeptidase levels remain stable during haemodialysis sessions on two different dialysis membranes. Finally, serum levels of the enzyme in anephric patients tend to be lower than in ESRF patients, suggesting that the kidney may contribute to the generation of the circulating form of endopeptidase 24.11. PMID- 2545996 TI - Effect of dietary trans fatty acids on some membrane-associated enzymes and receptors in rat heart. AB - Three groups of male weanling Sprague-Dawley rats were fed diets containing 20% corn oil, 20% partially hydrogenated soybean oil (PHSBO) or 18% PHSBO + 2% corn oil. PHSBO contained about 48% of its total fatty acids as trans-octadecenoate. Rats were killed after 16-18 weeks of feeding the various diets, hearts were dissected and crude sarcolemma was prepared by differential centrifugation. The activities of ouabain-sensitive (Na+ + K+)ATPase were significantly lower in membranes of rats fed 20% PHSBO than the control rats fed 20% corn oil. The feeding of 2% corn oil with 18% PHSBO resulted in partial restoration of the enzyme activity. The maximum number of [3H]ouabain-binding sites (Bmax) was also lower in cardiac membranes of rats fed 20% PHSBO than those fed 20% corn oil. Similar to (Na+ + K+)-ATPase activity, some restoration of the number of [3H]ouabain-binding sites was observed when 2% corn oil was fed with 18% PHSBO containing diet. There was no difference in the binding affinity of the radioligand for the receptor among the 3 dietary groups. Adenylate cyclase activities (fluoride-, isoproterenol- and forskolin-stimulated) were lower in membranes of rats fed 20% PHSBO or 18% PHSBO + 2% corn oil than in the control group fed 20% corn oil. Density of the beta-adrenergic receptor was the lowest in cardiac membranes of rats fed 20% PHSBO. The feeding of 2% corn oil with 18% PHSBO resulted in partial restoration of the maximum number of [3H]dihydroalprenolol (DHA)-binding sites.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2545995 TI - Phosphatidylinositol 4,5-bisphosphate phospholipase C activity in particulate preparations from rat brain. AB - We describe the hydrolysis of phosphatidylinositol 4,5-bisphosphate by a particulate rat brain preparation in the presence of the cationic detergent, cetrimide. The optimum cetrimide concentration was in the range 0.2-0.4 mg/ml; at higher or lower concentrations, the reaction rate diminished abruptly, suggesting that the electrical charge density of the micelle is critical for enzymatic attack. In other respects, such as its partial requirement for Ca++ and its pH optimum of about 7.0, the particulate enzyme seems similar to soluble preparations which have been reported previously. Interestingly, the particulate preparation could be stimulated about fourfold by a soluble brain extract in the presence of 1 mM guanosine triphosphate, confirming that the enzyme is the catalytic subunit in a membrane-bound signal-transduction system. PMID- 2545998 TI - Development of cavernous vasculatures in livers with hepatocellular carcinoma. An autopsy study. AB - The incidence and morphology of cavernous vasculatures in livers with hepatocellular carcinoma were examined. These vasculatures were grossly or microscopically found in 64% of 102 autopsied livers with hepatocellular carcinoma, though the incidence of grossly visible vasculatures was only 7%. These vasculatures were preferentially found within the portal tracts in the vicinity of portal veins obstructed by tumor emboli, especially in the cases with marked tumor emboli of the portal venous system. A majority of these vasculatures showed histochemical and immunohistochemical characteristics of blood vessels. Our findings suggest that these vasculatures frequently develop as collaterals of portal veins obstructed by the tumor emboli in hepatocellular carcinoma. PMID- 2545999 TI - Iron induction of ferritin synthesis and secretion in human hepatoma cell (Hep G2) cultures. AB - In order to determine if iron was able to stimulate specifically ferritin synthesis and secretion in transformed human hepatocytes in culture, human hepatoma cell (HepG2) cultures were submitted to increasing doses of ferric nitrilotriacetate. Iron uptake by the cells was demonstrated by incorporation of 59 Fe and the staining method of Perls. The following results were obtained: 1. iron incorporation within the hepatocytes increased as a function of culture time; 2. during the first 24 h of treatment, ferritin synthesis increased progressively, in parallel to the iron uptake; 3. a dose-dependent significant stimulation of ferritin synthesis and secretion were observed when the medium iron concentration increased from 5 to 20 mumol/l; 4. albumin, transthyretin and transferrin secretions were unaffected. These data demonstrated that, in our hepatocyte culture model, iron load increased the expression of ferritin in a highly specific manner. PMID- 2545997 TI - Eskimo plasma constituents, dihomo-gamma-linolenic acid, eicosapentaenoic acid and docosahexaenoic acid inhibit the release of atherogenic mitogens. AB - Studies in man and laboratory animals suggest that omega 3 polyunsaturated fatty acid constituents of fish oils have antiatherosclerotic properties. We have studied the effects of several such polyunsaturated fatty acids for ability to modify the in vitro release of mitogens from human platelets. Such mitogens may produce the fibro-proliferative component of atherosclerotic plaques. Both 5,8,11,14,17-eicosapentaenoic acid (20:5 omega 3) and 4,7,10,13,16,19 docosahexaenoic acid (22:6 omega 3), major constituents of fish oils, inhibited adenosine diphosphate-induced aggregation of platelets and the accompanying release of mitogens. These effects are dose dependent. Linolenic acid (18:3 omega 3), the biosynthetic precursor of eicosapentaenoic acid, also inhibited platelet aggregation and mitogen release. Eicosapentaenoic acid also inhibited mitogen release from human monocyte-derived macrophages, which, in vivo, are an additional source of mitogens during atherogenesis. Potent inhibition of human platelet aggregation and mitogen release was also seen with dihomo-gamma linolenic acid (8,11,14-eicosatrienoic acid 20:3 omega 6), whose levels are reportedly elevated in Eskimos subsisting on marine diets. We conclude that diets that elevate plasma and/or tissue levels of eicosapentaenoic acid, docosahexaenoic acid and dihomo-gamma-linolenic acid precursor gamma-linolenic acid (18:3 omega 6) may exert antiatherosclerotic effects by inhibiting the release of mitogens from platelets and other cells. PMID- 2546000 TI - Development of beta-adrenergic receptor and G protein messenger RNA in rat brain. AB - The ontogeny of rat brain beta 1- and beta 2-adrenergic receptor (beta-AR) and G protein messenger RNA (mRNA) was examined by Northern blot analysis using nick translated cDNA clones for probes. The level of beta 1-AR and beta 2-AR mRNA followed a pattern of development which paralleled that for the receptor binding sites; the levels of mRNA and binding sites were low at day 1 and increased gradually to adult levels by postnatal days 16-25. In contrast, the level of G protein mRNA, including that for Gs alpha, Gi1 alpha, Gi2 alpha, Go alpha and G beta, on postnatal day 1 was equal to or greater than adult levels, increased 40 80% between day 1 and 7 and then decreased to adult values by day 14-25. This developmental time course approximates that reported for the expression of Gs and Gi but not Go and G beta protein levels determined by immunolabeling and toxin catalyzed ADP-ribosylation. The level of beta-actin mRNA was also greater than adult levels on day 1 and then gradually decreased to adult levels by day 14-25. The results indicate that the ontogeny of beta-ARs and G proteins and the relationship between the amount of mRNA and protein are qualitatively different. PMID- 2546001 TI - Localization of GABAA receptor alpha-subunit mRNAs in relation to receptor subtypes. AB - The distribution of 3 GABAA receptor alpha-subunit mRNAs in various regions of bovine brain has been investigated using in situ hybridization. Whereas the alpha 2- and alpha 3-transcripts are of low abundance in all regions except striatum, the alpha 1-transcript is considerably enriched in the inferior colliculus, olfactory bulb and substantia nigra, and appears to be correlated with benzodiazepine type I receptor localization. PMID- 2546002 TI - The expression of the DNA ligase gene of Escherichia coli is stimulated by relaxation of chromosomal supercoiling. AB - Many genes of Escherichia coli have been shown to be sensitive to DNA superhelicity. The superhelicity of the chromosome is itself also supercoiling dependent. We have developed a general strategy for investigating how a particular gene responds to changes in DNA topology. This approach is used to study the E. coli ligase gene. The thermosensitivity of the E. coli ligts251 mutation can be phenotypically suppressed by mutations which map close to, or in, the gyrB gene and which affect the degree of DNA supercoiling. The level of suppression correlates with the degree of DNA relaxation observed, suggesting that the gene encoding the E. coli DNA ligase is activated by relaxation of the chromosomal DNA. PMID- 2546003 TI - Molecular cloning of the outJ gene involved in pectate lyase secretion by Erwinia chrysanthemi. AB - The phytopathogenic enterobacterium Erwinia chrysanthemi secretes a number of enzymes involved in plant-tissue degradation, notably the five isoenzymes of pectate lyase. We have cloned a region involved in pectate lyase and cellulase secretion by complementation of non-secretory outJ mutants of E. chrysanthemi strain 3937 using the RP4::miniMu plasmid pULB110. The cloned region maps near the ade-22 marker on the E. chrysanthemi 3937 chromosome. An R-prime containing a chromosomal DNA insert of about 30kb was first obtained; subcloning into pBR325 permitted the isolation of a 4kb ClaI/SspI fragment able to complement outJ mutations in E. chrysanthemi. The isolation of phoA fusions in this fragment allowed us to determine the direction of transcription of the encoding region, which extends over about 2.5kb, and demonstrate that this region encodes exported protein(s). When the TnphoA insertions were transferred back into E. chrysanthemi chromosome, the recombined strains no longer secreted pectate lyases or cellulases. Identification of the products encoded by the ClaI/SspI fragment demonstrated that outJ encodes an 83 kD polypeptide which is processed to an 81 kD polypeptide by cleavage of a signal sequence. The cloned DNA fragment did not endow Escherichia coli with the ability to secrete pectate lyases. PMID- 2546004 TI - Fine structural analysis of the Zoogloea ramigera phbA-phbB locus encoding beta ketothiolase and acetoacetyl-CoA reductase: nucleotide sequence of phbB. AB - A series of expression plasmids containing either the complete insert from plasmid pUCDBK1 (Peoples et al., 1987) or sub-fragments thereof were constructed in a tac promoter vector. Analysis of protein lysates of induced cultures of these clones identified the gene encoding NADPH-specific acetoacetyl-CoA reductase in the 2.3kb of sequence located downstream from the beta-ketothiolase gene in plasmid pUCDBK1. The complete nucleotide sequence (2.1kb) of this region was determined. An open reading frame was located 88bp downstream from the stop codon of the thiolase gene encoding a potential polypeptide of Mr 25,000, which is in good agreement with that observed for the overexpressed protein on SDS PAGE. N-terminal protein sequence data obtained by Edman degradation of the purified Mr = 25,000 polypeptide were used to identify the correct start of the NADPH-specific acetoacetyl-CoA reductase gene. Hence in Z. ramigera, the genes encoding beta-ketothiolase (phbA) and NADPH-specific acetoacetyl-CoA reductase (phbB) are organized as phbA-phbB. S1-nuclease analysis of Z. ramigera RNA identified a transcription start site 85 bp upstream from the phbA structural gene locating the promoter region. PMID- 2546005 TI - Phospholipase C and haemolytic activities of Clostridium perfringens alpha-toxin cloned in Escherichia coli: sequence and homology with a Bacillus cereus phospholipase C. AB - The Clostridium perfringens alpha-toxin (phospholipase C) gene (cpa) has been cloned and expressed in Escherichia coli. The biological activities of the cloned gene product have been analysed and the complete nucleotide sequence of the cpa gene has been determined. The cloned cpa gene product, which is exported to the periplasm in E. coli, possesses both phospholipase C and haemolytic activities. Haemolysis is not apparent when cell extracts are incubated with isotonic suspensions of sheep erythrocytes, but can be detected and quantified readily when dilutions of the same extracts are placed in wells in sheep-blood agar plates. Like other sequenced clostridial genes, the cpa gene has a high AT content (66.4%), exhibits a strong bias for using codons with A or T in the wobble position, and the 350 base pairs upstream from the gene have a significantly higher AT content (79.5%) than the coding region. The cpa gene encodes a 398 amino acid polypeptide with a deduced molecular weight of 45,481 D. This is very similar to the estimated molecular weight (Mr) of the cpa primary gene product expressed in an in vitro transcription-translation system (Mr 46,000), but larger than the cpa gene product detected in E. coli minicells, E. coli whole cells or in C. perfringens cells (Mr 43,000), suggesting post translational processing. The 28 N-terminal residues of the deduced alpha-toxin sequence possess the consensus features of a signal peptide and may be removed during secretion. The deduced alpha-toxin sequence shares significant structural homology with the phosphatidylcholine-preferring phospholipase C of Bacillus cereus. PMID- 2546006 TI - Gene structure and precursor processing of a novel Bacillus subtilis spore coat protein. AB - The gene for an unusual 8kD Bacillus subtilis spore coat polypeptide has been cloned and sequenced. It contains high percentages of proline, glycine and tyrosine, lacks thirteen amino acids, and is present as the carboxyl two-thirds of an open reading frame encoding a 12kD polypeptide. Two presumptive precursors which could be converted to the 8kD antigen by incubation with trypsin were found in extracts of cells or spores of a strain containing multiple copies of this gene. Large amounts of these coat antigens were also present in extracts of a germination-defective mutant which is altered in spore coat structure. There was little 8kD coat protein in the mutant, however, implying that processing is dependent on proper coat assembly. This gene was mapped to the metA region of the B. subtilis chromosome, a unique location as is true for other spore coat genes. Transcription and translation occurred late in sporulation (stage V) and the upstream region contained sequences similar to those found in other spore coat genes. PMID- 2546008 TI - Cloning and expression of plasmid DNA sequences involved in Salmonella serotype typhimurium virulence. AB - A 22kb region of the 90kb virulence-associated plasmid, pIP1350, of Typhimurium strain C52 was cloned into the mobilizable vector pSUP202, yielding plasmid pIP1352. This recombinant plasmid restored full virulence to plasmidless strain C53 in a mouse model. Transposon Tn5 insertion mutagenesis demonstrated the existence of two DNA sequences in pIP1352 designated VirA and VirB, both of which are essential for the expression of virulence. A recombinant plasmid containing only the VirA and VirB regions markedly increased the virulence of the plasmidless strain C53, but did not confer full virulence. These results suggested that a third virulence-associated region might be present on pIP1352. Eleven proteins encoded by the 22kb insert sequence of pIP1352 were identified in Escherichia coli SE5000 maxicells. The VirA region encoded at least two proteins with apparent molecular weights of 71,000 and 28,000 and the VirB region encoded two proteins of 43,000 and 38,000. PMID- 2546007 TI - Identification, molecular cloning and sequence analysis of a gene cluster encoding the class II fructose 1,6-bisphosphate aldolase, 3-phosphoglycerate kinase and a putative second glyceraldehyde 3-phosphate dehydrogenase of Escherichia coli. AB - To investigate a possible chromosomal clustering of glycolytic enzyme genes, the complete nucleotide sequence of the 8029 bp insert of Escherichia coli DNA in the ColE1 plasmid pLC33-5 of the Clarke and Carbon collection (Clark and Carbon, 1976) was determined. Genes (pgk, fda) encoding the phosphoglycerate kinase and Class II fructose 1,6-bisphosphate aldolase, respectively, of E. coli were identified. The phosphoglycerate kinase was found to be highly homologous in primary structure to the same enzyme from eukaryotic organisms. A further large open reading frame, designated gapB, was also identified, which on the basis of sequence homology, appears to encode another glycolytic enzyme, glyceraldehyde 3 phosphate dehydrogenase. This putative gene differs significantly from that (designated gapA) already identified as coding for this enzyme in E. coli and which maps elsewhere on the chromosome. The products, if any, of several other open reading frames remain to be identified. PMID- 2546009 TI - The Rhizobium meliloti host range nodQ gene encodes a protein which shares homology with translation elongation and initiation factors. AB - The Rhizobium meliloti nod region IIb is involved in host-range determination: (i) the presence of region IIb is necessary for transfer of alfalfa root hair curling ability to Rhizobium leguminosarum biovar trifolii; (ii) a mutation in region IIb extends the R. meliloti infection host range to Vicia sativa nigra; (iii) dominance of R. meliloti nod genes over R. leguminosarum biovar viciae nod genes is abolished by mutations in region IIb. The nucleotide sequence of this region has been determined. Genes corresponding to the two open reading frames identified are designated nodP and nodQ. The predicted amino acid sequence of the NodQ protein shows homology with translation initiation and elongation factors. The consensus sequence involved in the GTP-binding domain is conserved. PMID- 2546010 TI - Analysis of a cloned sequence of Legionella pneumophila encoding a 38 kD metalloprotease possessing haemolytic and cytotoxic activities. AB - The DNA encoding the zinc metalloprotease of Legionella pneumophila Philadelphia 1 has been isolated and expressed in Escherichia coli. This protein, which is 38,000 Daltons in size, possesses immunological and biochemical properties identical to those previously described for the purified L. pneumophila protease. Periplasmic extracts of E. coli clones expressing the recombinant protease are also capable of causing the haemolysis of canine erythrocytes and the cytotoxic destruction of CHO cells. Using transposon mutagenesis, it was determined that a maximum of 1.2 kb of DNA encoded all three biological activities. Inactivation of proteolytic activity by transposon insertion occurred concomitantly with losses of the haemolytic and cytotoxic phenotypes. A putative regulatory sequence approximately 200-500 bp upstream of the gene's coding region was identified. A 4.0 kb fragment encoding these activities hybridized to the chromosomal DNA of the parent strain of L. pneumophila Philadelphia 1 as well as clinical isolates of L. pneumophila. PMID- 2546011 TI - Analysis of C4-dicarboxylate transport genes in Rhizobium meliloti. AB - A 5.1 kbp DNA fragment was isolated which complemented C4-dicarboxylate transport mutants (dct) of Rhizobium meliloti. Characterization of this fragment by subcloning, transposon mutagenesis, and complementation analysis revealed three loci, designated dctA, dctB, and dctD. TnphoA-generated alkaline phosphatase fusions to dctA suggested that this gene encodes the structural transport protein and allowed the determination of its direction of transcription. Analysis of the fusions in various mutant backgrounds demonstrated that dctB, dctD, and ntrA products are required for dctA expression. The dctA fusion was constitutively expressed in a dctA mutant background, but was not expressed in dctA dctB or dctA dctD double mutants. This suggests that the constitutive expression in a dctA mutant background is mediated through dctB and dctD. Three independent second site Dct+ revertant mutations in ntrA mutant strains mapped to the dct locus. Succinate transport in these revertant strains was constitutive, whereas in the wild type, succinate transport was inducible. These results are consistent with the direct requirement of the ntrA gene product for dctA expression. Alfalfa plants inoculated with the dctB and dctD mutants showed reduced nitrogen-fixing activity. Nodules induced by dctA mutants failed to fix nitrogen. These symbiotic phenotypes are consistent with previous suggestions that dctA expression in bacteroids can occur independently of dctB and dctD. PMID- 2546012 TI - Preparation and use of micro- and macroelectrodes for measurement of transmembrane potentials and ion activities. PMID- 2546013 TI - Measurement of ion fluxes in membrane vesicles using rapid-reaction methods. PMID- 2546014 TI - Electron paramagnetic resonance methods for measuring pH gradients, transmembrane potentials, and membrane dynamics. AB - In this article, we have described the use of paramagnetic hydrophobic ions and secondary alkylamines to estimate pH gradients, and transmembrane and interfacial boundary potentials in membrane vesicle systems. Several features of these probes make them powerful tools for the investigation of membrane potentials. The mechanisms by which they function are well understood, thereby facilitating their use in a quantitative fashion. They have a relatively high sensitivity and are not affected by turbidity. While they are ideally suited for use in model membrane systems, they have also been used successfully in biological and reconstituted membrane systems. Finally, the synthetic flexibility of the nitroxide permits a much wider range of probes to be synthesized than has been described here. Future work will likely bring additional developments in the application and information obtainable from this methodology. PMID- 2546015 TI - Radiation inactivation of membrane components and molecular mass determination by target analysis. PMID- 2546016 TI - Membrane protein molecular weight determined by low-angle laser light-scattering photometry coupled with high-performance gel chromatography. PMID- 2546017 TI - Cross-linking techniques. PMID- 2546019 TI - Some theoretical aspects of the mechanism of action of benzhydrazone, an inhibitor of glycosylation in herpes simplex virus. AB - Some hypotheses of the mechanism of action of benzhydrazone, a selective inhibitor of herpes simplex virus glycoprotein biosynthesis, are presented in this paper. Comparative analysis of different models indicates that the drug may interfere with herpes simplex virus DNA. Thus, benzhydrazone seems to act like a bifunctional alkylating agent. PMID- 2546018 TI - Membrane-impermeant cross-linking reagents. PMID- 2546020 TI - A serologic survey of encephalomyocarditis virus infection in pigs in Italy. AB - Neutralizing antibodies to encephalomyocarditis virus (EMCV) were found in 69.13% of the serum samples obtained from pigs from representative regions of Italy. The antibodies are distributed fairly uniformly throughout the swine populations with all age groups being equally involved. PMID- 2546021 TI - A neuroanatomical and biochemical basis for attention deficit disorder with hyperactivity in children: a defect in tonic adrenaline mediated inhibition of locus coeruleus stimulation. AB - Attention deficit disorder with hyperactivity (ADDH) is characterized by a high level of inappropriate activity, distractability and impulsivity as well as learning deficits. In mammals, the protective mechanism for acquisition of sensory information, processed for its survival value, can be termed "vigilance". In ADDH, a low threshold for novel or sensory stimuli, results in a "hard wired" mammalian response, orienting, followed by exploratory behavior, the orienting reaction. This behavior is mediated in the brainstem through sensory neurons in the reticular formation regulating the discharge of locus coeruleus noradrenergic neurons. It is proposed that adrenaline acts as a tonic inhibitor of these neurons, modulating the level of sensory stimulation necessary to elicit locus coeruleus excitation. An imbalance in adrenaline formation or alpha-2 adrenergic receptor number (or affinity) leads to the inability to maintain the appropriate threshold for discharge of locus coeruleus neurons. The consequences are inability to maintain focused attention, difficulty in falling asleep or light levels of sleep, inattention to consumptive behavior, and probable inappropriate response for reward as all these behaviors require disruption of processing of sensory stimuli. Cognitive deficits may occur secondarily. PMID- 2546022 TI - Inhibiting noradrenergic overactivity by inhibition of thromboxane and concomitant activation of opiate receptors via dietary means. AB - A yin-yang hypothesis is presented linking noradrenergic activity, thromboxane, melatonin, left hemisphere functioning, and cyclic AMP on the one hand, and dopamine, beta-endorphin, calcium, right hemisphere functioning, and cyclic GMP on the other. It is further suggested that there is a yoking of NA, TXA2, serotonin and melatonin in the left hemisphere, and a similar yoking of DA, BE, calcium and cGMP in the right. Evidence is presented to support the hypothesis that each element (NA, TXA2, etc.) on one side can modulate or balance a corresponding element (DA, BE, etc.) on the other. It is suggested that thromboxane is the key element in noradrenergic overactivity and that not taking this into consideration has confounded much prior research. This theory takes into account information processing models as well as pharmacological data and neurochemical theory on coupling of adenylate cyclase to its hormone receptors. Inhibiting noradrenergic overactivity can be obtained by inhibiting thromboxane and concomitantly activating opiate receptors. This protocol may have clinical utility in treating a wide range of disorders such as: anxiety, depression, schizophrenia, sleeplessness, withdrawal states, enuresis, Gilles de la Tourette syndrome, Parkinsonism, Alzheimers, dementia, anorexia, infant ruminations, essential tremor, spasticity of spinal cord injury, diarrhoea, ulcerative colitis, extrapyramidal symptoms, akathisia, neuroleptic malignant syndrome, attention deficit disorder, hyperhidrosis, and possibly AIDS. PMID- 2546023 TI - Cytomegalovirus detection by biotinylated DNA probes. AB - Clinical specimens from 317 patients suspected of cytomegalovirus infection were examined by immunofluorescence (IF) using monoclonal antibodies and by a biotinylated DNA probe kit after cell culture isolation. Of the 317 samples, 68 were positive by culture isolation. Of these 67 were IF positive when the cytopathic effect (CPE) was 1+ or less, whereas 56 gave positive results with DNA probes when the CPE was 2+. A further 83 specimens were examined directly by immunoperoxidase histopathology (IHP), IF and the DNA probe kit: 26 of these were positive by IHP examination, 25 by IF and only 6 by DNA probes. The sensitivity of the DNA probe kit was not satisfactory when the clinical tissue specimens were directly examined. However, the sensitivity improved considerably to 82% if the specimens were propagated first in cell culture. The IF method detected the virus before and after cell culture isolation equally well (96%-98.5%). Compared to the IF method, the DNA probe kit is costly and requires more labor and time. PMID- 2546025 TI - Multidisciplinary treatment of metastatic germ cell tumor: clinical summary and discussion. PMID- 2546024 TI - Biological function of the rotavirus protein VP4: observations on porcine isolates from China. AB - Rotaviruses isolated from pigs in China were grown in MA104 cells. One tissue culture-adapted isolate consisted of two subpopulations (variants), the RNA profiles of which differed in the relative migration of RNA segment 4 only. The variants were separated by plaque purification and by recovery from limiting dilutions and remained genetically stable. The variant possessing the slower migrating RNA segment 4, called 4S, grew faster and formed large plaques after 4 6 days incubation, whereas the variant possessing the faster migrating RNA segment 4, called 4F, grew more slowly and formed only microscopic plaques after 10-14 days incubation. The protein product of the 4F RNA occurred in much lower concentration in infected cells than the product of the 4S RNA. The RNA segments 4 of the two variants were found to be closely related when tested by dot hybridization under stringent conditions. The 4S RNA is more resistant to denaturation with methyl mercuric hydroxide than is the 4F RNA. The relevance of these findings to the biological functions of rotaviruses is discussed. PMID- 2546028 TI - Ductal carcinoma in situ of the breast. PMID- 2546027 TI - Prognostic significance of Auer rods in childhood acute myelogenous leukemia: results of the studies AML-BFM-78 and -83. AB - The prognostic significance of Auer rods in predicting response rate and remission duration was investigated in 257 patients of the two West German acute myelogenous leukemia (AML) studies BFM-78 and -83. Auer rods were found in 129 children (50%) in the initial bone marrow smear. The incidence was higher in myeloid subtypes M1 (63%) and M2 (78%) compared with subtypes M4 (47%) and M5 (5%) with monocytic differentiation. In both studies, the remission rate was significantly higher in patients with Auer rods (P = 0.01), and in the study AML BFM-83 a significantly longer remission duration was evaluated for Auer rod positive patients (P = 0.007). In the M1 type, Auer rods were of high prognostic significance regarding both the induction success (P = 0.003) and the remission duration (P = 0.004), whereas no significant differences between Auer rod positive and -negative patients in other subtypes were found. Occurrence of Auer rods was independent of hyperleukocytosis, the most powerful prognostic parameter in the studies AML-BFM-78 and -83, whereas absence of Auer rods was associated with the AML risk group M5. In the M1 type, Auer rod-negative leukemias appears to represent cases of poorly differentiated AML. Auer rods were therefore useful in differentiating between patients with a poor and a more favorable prognosis, particularly in the M1 type. PMID- 2546026 TI - Phase II study of N-methylformamide, spirogermanium, and 4-demethoxydaunorubicin in the treatment of non-small cell lung cancer (EST 3583): an Eastern Cooperative Oncology Group study. AB - One hundred forty-four patients with non-small cell lung cancer, the majority (72%) of whom had received previous chemotherapy, were evaluable in this randomized phase II study of N-methylformamide (N-MF), spirogermanium, and 4 demethoxydaunorubicin. There were two partial responses, one each with spirogermanium and 4-demethoxydaunorubicin. There were eight life-threatening complications (mostly hematologic) and two lethal complications (N-MF, hematologic; 4-demethoxydaunorubicin, gastrointestinal). The overall survival ranged from 9 days to 533 days with a median of 17.6 weeks. The following factors were associated with poor survival: Poor initial performance status, prior weight loss, presence of liver or subcutaneous metastases. PMID- 2546029 TI - Hepatocellular carcinoma with tumor thrombus in inferior vena cava. PMID- 2546030 TI - Response to Drs. Pritchard and Pinkerton. PMID- 2546031 TI - [Intracellular uptake and accumulation of metaiodobenzylguanidine in neuroblastoma cells and small cell carcinoma of the lung]. AB - Recently m-131-iodobenzylguanidine (MIBG) has been successfully used to image neurocrest-derived tumours such as neuroblastoma. In fact, our autoradiographic evidences demonstrate that MIBG is able to bind, accumulate and be released by neuroblastoma cell line CHP 100 "in vitro", as well as by surgical biopsies from tumours bearing children. Moreover, all the cell lines tested "in vitro" were able to bind MIBG: neuroblastoma (WNT, CHP-100, IMR-32), small cell lung carcinoma (LC-8) and melanoma (WMB). A fibrosarcoma line, despite that it is not neurocrest related, was also able to bind MIBG. However only WNT, CHP-100 and the LC-8 lines were able to accumulate and then release large amounts of MIBG in cytoplasmatic vesicles. The heterogeneity of MIBG uptake "in vitro" seems related to the similar variability observed in the clinical scintigraphies. This phenomenon may be related to the biochemical maturity of individual neuroblastoma tumours. PMID- 2546032 TI - [Treatment of chronic functional constipation in children by administration of vegetable fiber (Dicoman 5)]. AB - Chronic functional constipation is a common clinical disorder in pediatric age. Thirteen children with functional constipation were treated by administration of vegetable fibres together with a toilet training program, for two months. In each patient anorectal manometry showed presence of inhibitory anal reflex and diagnosis of constipation was confirmed by a prolonged gastrointestinal transit time measured by radio-opaque markers. In all patients there was a significant improvement in both stool frequency and intestinal transit time; furthermore, a normalization of anorectal motility variables was observed at rectal manometry. No changes in the blood levels of nutritional parameters were seen in any patient. It is concluded that vegetable fibres represent an effective treatment of functional chronic constipation in children. PMID- 2546033 TI - [Biological activity of hepatocellular carcinoma by analysing nuclear DNA ploidy patterns and using anti BrdU monoclonal antibody]. AB - Cell nuclear DNA ploidy patterns were examined using cytofluorometry in hepatocellular carcinomas (HCC) induced by diethylnitrosamine (DEN) in rat and human HCC. These ploidy patterns were compared with histopathological and/or immunohistochemical studies of HCC using anti-bromodeoxyuridine monoclonal antibody. The results are summarized as follows. HCC of rats induced by DEN was occurred in 74(65.5%) of 113 rats. Intravascular invasion of the livers were seen in 31 of 74(41.9%). Metastasis was in 25 of 74(33.8%). The ploidy patterns of HCC in rats were classified into three types (types I, II and III). These ploidy patterns were closely connected with histological types of HCC in Edmondson's classification, and in order of types changing from type I to type III, 6c over polyploid cells, mitotic indices (MI) and labeling indices (LI) had a tendency to increase. In human HCC, cell nuclear DNA ploidy patterns were classified into three types similarly to experimental HCC. These ploidy patterns were closely related to histological types in Edmondson's classification. MI and 6c over polyploid cells were tendency to increase similarly to HCC of experimental rats. From the above results, measurement of DNA ploidy patterns and LI are considered a valuable parameter in examining the biological activity of HCC. PMID- 2546034 TI - [Experimental study of hepatic hemodynamics and hepatic oxygen consumption after hepatectomy]. AB - This study was undertaken in order to clarify the hepatic hemodynamics and hepatic oxygen consumption in the remaining liver after 30% and 70% hepatectomy using adult mongrel dogs and moreover was investigated the improving effect of drugs (dobutamine (DOB), dopamine (DA), dibutyryl cyclic AMP (DBcAMP) and SS-094) after hepatectomy. The hepatic blood flow after hepatectomy decreased depending on the percentage of the liver resected but the hepatic blood flow per unit liver weight increased. The hepatic oxygen consumption also decreased but the hepatic oxygen consumption per unit liver weight increased, particularly after massive hepatectomy, resulting in the elevation of hepatic oxygen extraction ratio. And the uptake of lactate per unit liver weight decreased after massive hepatectomy. The administration of DOB, DA, DBcAMP and SS-094 after massive hepatectomy increased the hepatic arterial flow and portal venous flow and as a result the hepatic oxygen delivery was increased. The improvement of hepatic hemodynamics resulted in normalizing of the relation between hepatic oxygen demand and supply, and improvement of the uptake of lactate in the remaining liver. Therefore it was considered that those drugs favored the remaining hepatic hemodynamics and metabolisms after massive hepatectomy. PMID- 2546035 TI - [Effect of activation of the reticuloendothelial system on hepatocyte regeneration in partially hepatectomized rats]. AB - Enhanced effect on rat hepatocyte regeneration through activation of the reticuloendothelial system (RES) was investigated. RES was activated using by OK 432 and the phagocytic activity (K-value) increased 2-fold over control rats 24 hours after intraperitoneal injection of OK-432. In OK-432 treated rats, the amount of cyclic AMP in the regenerating liver also increased 1.5-fold higher than in control rats 14 hours after hepatectomy. After 70% partial hepatectomy, liver DNA synthesis in OK-432 treated rats increased 2 to 5-fold higher than in control rats. There was a good correlation between the K-value before hepatectomy and DNA synthesis 24 hours after hepatectomy (r = 0.96). Therefore the RES is one of the most important regulatory factors in liver regeneration. Augmentation of RES before hepatectomy may prevent hepatic failure after resection in liver diseases. PMID- 2546036 TI - [Malignant fibrous histiocytoma of the liver]. AB - Primary sarcomas of the liver are rare malignant neoplasms. Only four cases of Malignant fibrous histiocytoma (MFH) of the liver have previously been reported in the literature. The authors report a 64-year-old man of MFH of the liver, complained of epigastric discomfort. Laboratory values revealed raised ESR and positive CRP. An abdominal computed tomographic scan disclosed 7.4 x 7.6 cm low density area in the posterior segment of the right lobe. Selective angiography of the celiac artery revealed hypovascular tumor. The diaphragm was involved in the tumor and right hepatectomy with partial resection of diaphragm was performed. Microscopically, the tumor cells were arranged with a storiform pattern and electron microscopic studies demonstrated fibroblast like cells and histiocyte like cells. Although systemic chemotherapy with ACR, CPM and VCR was performed, he died of peritoneal and pleural dissemination on the 142nd day after. PMID- 2546037 TI - Transcriptional occlusion of transposon targets. AB - In Salmonella typhimurium, insertion of transposons Tn5, Tn10 and bacteriophage Mu is inhibited by transcription of some target sequences. The transcription effects on Tn5 are large when the lac operon is a target but are limited to a slight effect on the hisG gene of the his operon. The Tn10 element shows target occlusion in both operons. Phage Mu has been shown previously to be inhibited for insertion into the lac operon. In the his operon Mu is only inhibited for insertion into the hisG gene. The variability of the inhibition effect from one sequence to another suggests site or regional specificity for transcription effects. Reducing the probability of insertion into transcribed sequences may be of selective importance to transposons since it reduces the risk of killing the host while maintaining the ability to transpose. PMID- 2546039 TI - dam methylation and the initiation of DNA replication on oriC plasmids. AB - Plasmid DNA containing the replication origin of the Escherichia coli chromosome (oriC) has been shown to be inefficient as a template for DNA synthesis in vitro when isolated from dam mutants. Here, we extend this study to hemimethylated oriC plasmids and to replication in dam-3 mutant enzyme extracts. The results show that: (1) hemimethylated oriC plasmids replicate with the same low efficiency as nonmethylated DNA; (2) DNA synthesis starts at oriC regardless of the methylated state of the template; (3) replication in dam-3 enzyme extracts is inefficient because this strain is deficient in DnaA protein; and (4) consistent with this observation, the copy number of the oriC plasmid pFH271 is reduced in the dam-3 mutant. However, we have found that low DnaA protein levels in dam-3 mutants are not sufficient to explain the reduced transformation efficiency of oriC plasmids. We suggest that there must exist in vivo inhibitory factors not present or present in low quantities in vitro which specifically recognize the hemimethylated or nonmethylated forms of the oriC region. PMID- 2546038 TI - Absence of insertions among spontaneous mutants of Salmonella typhimurium. AB - While insertion sequences (IS) in Escherichia coli transpose frequently to generate spontaneous insertion mutants, such mutations are rare in Salmonella typhimurium: the only documented insertion mutation is a hisD mutation caused by the Salmonella-specific IS element IS200. To obtain more examples of IS200 insertion mutations and to seek additional types of IS elements in Salmonella, we selected and characterized 422 independent, spontaneous His- mutants and some 2100 additional mutants that are not necessarily independent. None of the mutants showed the absolute polar effect characteristic of insertion mutations or the reversion properties characteristic of insertions (low spontaneous reversion frequency and no reversion induction by chemical mutagens). A few mutants, showing a high spontaneous reversion frequency, were screened physically. No insertion mutations were found. Thus insertion mutations appear to be rare in S. typhimurium, in strong contrast to E. coli and despite the possession in Salmonella of at least one type of insertion element (IS200). These results suggest that in Salmonella transposition of the endogenous elements has been controlled. The transposition ability of the elements may have been reduced or favored target sites removed from the host genome. PMID- 2546040 TI - Cloning and DNA sequence of plasmid determinant iss, coding for increased serum survival and surface exclusion, which has homology with lambda DNA. AB - Escherichia coli K12 cells carrying a cloned 1.4 kb HindIII fragment from plasmid ColV2-K94, showed increased survival in guinea pig serum. The recombinant plasmid also conferred group II surface exclusion, i.e. the cells were reduced in recipient ability towards the incoming plasmid R538drd in conjugation experiments. Southern blotting suggested homology with bacteriophage lambda DNA and to the insertion element IS2. Determination of the DNA sequence of the fragment demonstrated the presence of a truncated IS2 (165 bp), separated by 250 bp from a 900 bp stretch of homology with lambda DNA, beginning within the Rz gene and continuing in the rightward direction on the lambda map. A 97 amino acid open reading frame (ORF) adjacent to Rz and on the opposite strand, remained intact in iss, with several amino acid changes. The ORF in iss is preceded by sequences resembling prokaryotic ribosome binding sites and promoters. PMID- 2546041 TI - Nucleotide sequence, evolutionary origin and biological role of a rearranged cytokinin gene isolated from a wide host range biotype III Agrobacterium strain. AB - A DNA fragment with homology to the cytokinin (ipt) gene from biotype I Agrobacterium tumefaciens strain Ach5 was cloned from the Ti plasmid of the wide host range biotype III Agrobacterium strain Tm-4 and sequenced. The fragment contains an intact ipt coding sequence. However, the 3' non-coding region of this ipt gene is rearranged due to a 0.9 kb deletion fusing it to the 3' coding region of the neighbouring gene 6a, most of which was found to be deleted. The Tm-4 ipt gene is strongly related to the partially deleted ipt gene of the limited host range biotype III strain Ag162. To test its biological activity, the Tm-4 ipt gene was inserted into a specially constructed, disarmed Ti vector lacking tzs and tested on tobacco, where the rearranged ipt gene induced shoot formation. The cloned Tm-4 ipt gene was mutated with Tn5 and the intact gene on the wild-type Tm 4 Ti plasmid was replaced by the mutated gene. The resulting strain was avirulent on tobacco but normally virulent on the natural host of the wild-type strain Tm 4, grapevine. As the biotype I 6b gene diminishes the effect of a corresponding ipt gene, a larger Tm-4 fragment carrying both the ipt gene and an adjacent 6b like gene was also tested on tobacco and compared with the Tm-4 ipt fragment alone and with an ipt and 6b/ipt fragment derived from Ach5. The Tm-4 6b gene diminishes the effect of the Tm-4 ipt gene, showing the Tm-4 6b gene to be active as well. The Tm-4 6b/ipt combination is less effective than the Ach5 combination. These results provide further insight into the molecular basis of the host range differences between limited host range and wide host range biotype III Agrobacterium strains and show that the WHR cytokinin gene, although active, does not significantly contribute to tumour formation on the natural host of the WHR biotype III strains, grapevine. PMID- 2546044 TI - Evidence for a pulmonary B3 bradykinin receptor. AB - We have examined pulmonary effects of bradykinin (Bk) in vivo and in vitro in guinea pigs and their potential inhibition by antagonists of Bk B1 and B2 receptors. Bk was a potent bronchoconstrictor in vivo and caused contractions of isolated, epithelium-denuded trachealis. D-Arg[Hyp3,D-Phe7]-Bk (NPC567) and D arg[Hyp3,Thi5,8,D-Phe7]-Bk (NPC349), B2 receptor antagonists, were weak inhibitors of Bk-induced bronchoconstriction in vivo and were virtually inactive as antagonists of Bk-induced airway smooth muscle contraction. Several other B2 antagonists as well as B1 antagonist, des-Arg9-[Leu8]-Bk, did not inhibit Bk induced tracheal contraction. The B1 receptor agonist des-Arg9-Bk was without effect on tracheal tone. Tracheal responses to Bk were unaffected by antagonists of muscarinic, histamine, serotonin, and catecholamine receptors. The inability of the antagonists to inhibit Bk is unlikely to be due to their degradation, because NPC567 was only weakly active in the presence of inhibitors of kininase I (EC 3.4.11.2), kininase II (EC 3.4.15.1), and neutral endopeptidase (EC 3.4.24.11). These studies were corroborated by ligand binding experiments in guinea pig and ovine airways. In [3H]Bk binding, the Bk antagonists had no effect in guinea pig trachea, slightly displaced [3H]Bk in ovine trachea, and inhibited approximately 60% of total specific binding in lung. des-Arg9-[Leu8]-Bk and several other agents, including atropine, neurokinin A, substance P, and vasoactive intestinal peptide, had no effect on lung Bk binding. Bk and its analogs were not degraded during the binding assay. These data suggest that pulmonary tissue, particularly in the large airways, contains a novel Bk binding site, a B3 receptor, which may be involved in Bk-induced bronchoconstriction. PMID- 2546042 TI - The effect of DnaA protein levels and the rate of initiation at oriC on transcription originating in the ftsQ and ftsA genes: in vivo experiments. AB - The DnaA protein of Escherichia coli, essential for initiation at oriC, binds at a defined sequence which occurs at the chromosomal origin, near plasmid replication origins and in the promoters of the dnaA and mioC genes. This sequence also occurs at many other sites on the E. coli chromosome including three sites within the essential cell division genes ftsQ and A. Using an fts-lac fusion phage, lambda JFL100, we show here that fts gene expression responds both to reduced and increased intracellular levels of DnaA protein in a manner consistent with the hypothesis that DnaA protein regulates fts gene expression. Experiments using dnaC and dnaB-ts strains, however, suggest that DnaA control of fts transcription may be indirect, at least in part, with fts responding to the rate of initiation at oriC as well as to changes in DnaA protein level per se. It differs in this respect from dnaA gene expression which is unaffected when initiation of replication is inhibited by DnaB or DnaC inactivation. Strains integratively suppressed with pKN500 behave anomalously; neither fts nor dnaA transcription is significantly increased when DnaA is inactivated in these strains. PMID- 2546043 TI - The PEP: fructose phosphotransferase system in Salmonella typhimurium: FPr combines enzyme IIIFru and pseudo-HPr activities. AB - We have cloned the fru operon of Salmonella typhimurium, coding for the enzymes of the phosphoenolpyruvate: fructose phosphotransferase system (Fructose PTS). The fruFKA operon consists of three genes: fruF coding for FPr, fruK for fructose 1-phosphate kinase and fruA for Enzyme IIFru. Insertions of Tn5 in the different genes were isolated and the activities of the gene products were measured. Expression of the plasmid-encoded fru operon in the maxicell system resulted in the synthesis of three proteins with molecular weights of 47 kDa (fruA), 39 kDa (fruF) and 32 kDa (fruK). We have sequenced the fruF gene and the regulatory region of the fru operon. In contrast to previously published results, we have found that the fruF gene codes for a 39 kDa protein, FPr, that combines Enzyme IIIFru and pseudo-HPr activities. The N-terminal part of FPr is homologous to the cytoplasmic domain of the Escherichia coli Enzyme IIMtl, as well as several Enzymes IIIMtl from gram-positive bacteria. The C-terminal domain shows homology to HPr of E. coli and several gram-positive organisms. The fru operon is regulated by a repressor, FruR. We have constructed an operon fusion between fru and the galK gene and shown that regulation of the fru operon by FruR takes place at the level of transcription. PMID- 2546045 TI - [125I]2-iodo-3,7,8-trichlorodibenzo-p-dioxin-binding species in mouse liver induced by agonists for the Ah receptor: characterization and identification. AB - The admininistration of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) to C57BL/6J mice produces a dose-related increase in the hepatic uptake of [125I]2-iodo-3,7,8 trichlorodibenzo-p-dioxin ([125I]Cl3DpD) in vivo and the binding of the radioligand to liver homogenate in vitro [Mol. Pharmacol. 36: 121-127 (1989)]. The TCDD-induced hepatic binding species was found to be predominantly in the microsomal fraction and was inactivated by heating at 60 degree, trypsin, and mercurials. The TCDD-induced binding species was found to have an apparent equilibrium dissociation constant, KD, ([125I]Cl3DpD) of 56 +/- 16 nM and a pool size, Bmax, of 22 +/- 5 nmol/g of liver. A number of halogenated dibenzo-p dioxins, biphenyls, and polycyclic aromatic hydrocarbons compete with [125I]Cl3DpD binding to this species; all are aromatic and planar. The distinctive profile of this binding species, a protein of large pool size induced in the microsomal fraction of liver but not other tissues and induced by agonists for the Ah receptor, suggested that this moiety might be cytochrome P3-450. The coincidence of the major microsomal species covalently labeled with the photoaffinity ligand [125I]2-iodo-3-azido-7,8-dibromodibenzo-p-dioxin and that immunochemically stained with polyclonal antiserum that binds to cytochrome P3 450 confirms this hypothesis. This is a novel role for a cytochrome P-450 isozyme, as an induced sequestration site that enhances the hepatic localization of the agonist drug. PMID- 2546046 TI - Stimulation of in vivo hepatic uptake and in vitro hepatic binding of [125I]2 lodo-3,7,8-trichlorodibenzo-p-dioxin by the administration of agonist for the Ah receptor. AB - [125I]2-lodo-3,7,8-trichlorodibenzo-p-dioxin ([125I]Cl3DpD), a radiolabeled, isosteric, analogue of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), was synthesized and used to study in vivo tissue localization and in vivo tissue binding. Twenty-four hours after the administration of a tracer dose (1 X 10(-10) mol/kg) of [125I] Cl3DpD to C57BL/6J mice, the hepatic concentration of radioactivity was 1-2% of the administered dose, whereas in mice pretreated with TCDD (1 X 10(-7) mol/kg), the hepatic accumulation of radiolabel was 25-30% of that administered. Liver homogenate from TCDD-treated mice bound 4 to 10 times more [125I]Cl3DpD than homogenate from control mice. The enhancement of in vivo uptake and in vitro tissue binding of [125I]Cl3DpD by TCDD administration was confined to liver and was not observed in other tissues examined, kidney, lung, spleen, small intestines, and muscle. The administration of TCDD to C57BL/6J mice produces dose-related stimulation of in vivo hepatic uptake of [125I]Cl3DpD, binding of radioligand to liver homogenate, and hepatic aryl hydrocarbon hydroxylase activity, with the dose for half-maximal stimulation, ED50, varying from 1.5 to 4.0 x 10(-9) mol/kg. In congenic C57BL/6J (Ahd/Ahd) mice, which express the lower affinity Ah receptor, the ED50 values for all three responses were shifted to approximately 10-fold higher doses. 3,3',4,4',5,5' Hexabromobiphenyl, a weak agonist for the Ah receptor produced a dose-related stimulation of these three responses in C57BL/6J mice (ED50 values of approximately 5 X 10(-7) mol/kg), but was without effect in C57BL/6J (Ahd/Ahd) mice. Stimulation of vivo hepatic uptake and in vitro liver homogenate binding of [125I]Cl3DpD was produced by administration of Ah agonists, such as 2,3,7,8 tetrachlorodibenzofuran and beta-naphthoflavone, but inactive congeners and other compounds that do not act via the Ah receptor, e.g. phenobarbital and pregnenolone-16 alpha-carbonitrile, did not evoke these effects. Thus, TCDD and other Ah agonists act through the Ah receptor to increase a liver binding species that increases the hepatic uptake of [125I]Cl3DpD in vivo and binding of this radioligand to liver homogenate in vitro. PMID- 2546047 TI - Thrombin and trypsin act at the same site to stimulate phosphoinositide hydrolysis and calcium mobilization. AB - Thrombin stimulates polyphosphoinositide hydrolysis in embryonic chick heart cells and in 1321N1 astrocytoma cells and increases intracellular Ca2+ in the 1321N1 cells. The serine protease trypsin mimics these actions in a dose dependent fashion, whereas the proteolytically inactive thrombin derivatives diisopropyl fluorophosphate-thrombin (DIP-thrombin) and D-phenylalanyl-L-prolyl-L arginine chloromethyl ketone-thrombin (PPACK-thrombin) are ineffective in this regard. The phosphoinositide responses to thrombin or trypsin and the muscarinic agonist carbachol are additive, but no additivity is observed between the responses to thrombin and trypsin. Unlike the response to carbachol, the phosphoinositide and Ca2+ responses to thrombin and trypsin desensitize, with no recovery of the calcium response even when Ca2+ stores are replenished. Cross desensitization of phospholipase C activation and calcium mobilization between these proteases is also observed. In addition, PPACK-thrombin, which elicits no response itself, effectively inhibits trypsin-stimulated phosphoinositide hydrolysis. It is proposed that thrombin and trypsin act through the same receptor. Proteolysis appears to be important in the mechanism by which these agonists elicit phosphoinositide hydrolysis, calcium mobilization, and, perhaps, subsequent receptor desensitization. PMID- 2546048 TI - Class I antiarrhythmic drug receptor: biochemical evidence for state-dependent interaction with quinidine and lidocaine. AB - The state-dependent binding of class I antiarrhythmic drugs to a receptor associated with the cardiac sodium channel was assessed using [3H]batrachotoxinin A 20-alpha-benzoate [( 3H]BTXB) binding. [3H]BTXB binds specifically to and stabilizes activated states of the sodium channel. Quinidine (IC50 = 40 microM) and lidocaine [IC50 = 61 microM) inhibited equilibrium [3H]BTXB binding to sodium channels present on freshly isolated rat cardiac myocytes. Scatchard analysis of [3H]BTXB binding in the presence of quinidine and lidocaine revealed two apparent patterns of inhibition. Quinidine (33 microM) increased the KD but had no significant effect on the Bmax, whereas lidocaine (91 microM) reduced the Bmax but had no significant effect on the KD. To address drug binding to activated and nonactivated states, we exploited the state-specific binding of [3H]BTXB. Drugs that increase the rate of dissociation (k-1) of [3H]BTXB must bind to sodium channels to which [3H]BTXB is already bound (i.e., activated channels). Therefore, drug-mediated increases in k-1 measure drug binding to activated states. Both quinidine and lidocaine increased the k-1 of [3H]BTXB, indicating drug binding to and destablization of activated sodium channels. However, the minimal affinities of quinidine and lidocaine for activated channels (KDact) were estimated to be 433 and 455 microM, respectively, concentrations much higher than the equilibrium IC50 values. Drugs that allosterically decrease the rate of association (k+1) of [3H]BTXB must bind to sodium channels to which [3H]BTXB is not already bound (i.e., nonactivated channels). Therefore, drug-mediated decreases in k+1 measures drug binding to nonactivated states. Quinidine and lidocaine decreased the k+1 of [3H]BTXB, indicating drug binding to and stablization of nonactivated sodium channels. The affinity of quinidine and lidocaine for nonactivated channels (KDnon) was estimated to be 10 and 35 microM, respectively, concentrations close to the equilibrium IC50 values. The markedly different KDact and KDnon values for both quinidine and lidocaine indicate state dependent binding of quinidine and lidocaine to the class I receptor on the cardiac sodium channel. Both drugs destabilize activated channels and stabilize nonactivated channels. The Scatchard results suggest that quinidine and lidocaine may have different mechanisms of allosteric inhibition of [3H]BTXB binding. PMID- 2546049 TI - Glycogen phosphorylase activation by two different alpha 1-adrenergic receptor subtypes: methoxamine selectively stimulates a putative alpha 1-adrenergic receptor subtype (alpha 1a) that couples with Ca2+ influx. AB - We compared the effects of methoxamine on alpha 1-adrenergic receptor-mediated phosphorylase activation in rat hepatocytes and rabbit aorta. Although methoxamine is a potent agonist in activating phosphorylase of rabbit aorta, it had little effect in rat hepatocytes. Using the phenoxybenzamine inactivation method, we found that the quantitative relationship between 125I-BE2254 (125I-BE) binding capacity and maximal norepinephrine-stimulated phosphorylase activation was nonlinear in rabbit aorta, whereas it was linear in rat hepatocytes. The potency of methoxamine in inhibiting specific 125I-BE binding is significantly (p less than 0.05) higher in rabbit aorta (Kd, 96.4 +/- 7.7 microM), compared with rat hepatocytes (Kd, 283 +/- 16 microM). However, these quantitative differences could not fully explain the blunted [Ca2+]c and phosphorylase responses to methoxamine in rat hepatocytes. Treatment with chlorethylclonidine dose dependently suppressed 125I-BE binding sites and norepinephrine-induced phosphorylase activation in rat hepatocytes, whereas in rabbit aorta it resulted in only a 31% decrease in 125I-BE binding sites, with little effect on phosphorylase activation. Furthermore, alpha 1-adrenergic receptor-mediated cellular events of phosphatidylinositol (PI) hydrolysis and phosphorylase activation were unaffected by the removal of extracellular Ca2+ in rat hepatocytes, whereas both responses were markedly attenuated in rabbit aorta. The results indicate that two different alpha 1-adrenergic receptor subtypes activate glycogen phosphorylase, through different mechanisms for increasing [Ca2+]c in the two systems. In rat hepatocytes, alpha 1 receptors are closely linked to PI hydrolysis and Ca2+ release from intracellular stores and cause phosphorylase activation. In rabbit aorta, on the other hand, activation of alpha 1 receptors increases [Ca2+]c by Ca2+ influx from the extracellular fluid as well as by Ca2+ release, and both PI hydrolysis and phosphorylase activation are caused mainly by the Ca2+ entry. Methoxamine interacts with both chlorethylclonidine-sensitive and -insensitive alpha 1 receptor subtypes but selectively stimulates the alpha 1 receptor subtype that closely couples with the Ca2+ influx. PMID- 2546050 TI - Comparison of two putatively selective radioligands for labeling central nervous system beta-adrenergic receptors: inadequacy of [3H]dihydroalprenolol. AB - [3H]Dihydroalprenolol ([3H]DHA) has been used extensively in receptor binding studies to measure beta-adrenergic receptors in the central nervous system. Usually, nonspecific binding has been defined by high concentrations of the beta adrenergic receptor agonist isoproterenol or antagonists such as alprenolol or propranolol. Scatchard plots of such "specific" [3H]DHA saturation data in rat cerebral cortex membranes are linear. However, computer analysis demonstrated that the competition curves of these drugs for 2.0 nM [3H]DHA binding are biphasic, with a continuous inhibition of [3H]DHA binding in the concentration range usually used to determine nonspecific binding. These data indicate that another saturable high affinity site was being labeled by the radioligand and that the definition of nonspecific binding with any of these unlabeled drugs is not satisfactory. We used the nonlinear, least squares, curve-fitting program LIGAND to analyze total [3H]DHA binding, allowing the program to mathematically define nonspecific binding as a function of 3H-ligand concentration. Significantly lower Bmax (-44%) and Kd (-58%) values for beta-adrenergic receptors were found, indicating that under normal experimental procedures (defining [3H]DHA non-specific binding with these nonradioactive drugs) a second binding site was being labeled. We found that [3H]DHA binding to this site could be inhibited by drugs such as RU24969, a 5-hydroxytryptamine1A (5HT1A) and 5HT1B receptor subtype-selective agonist, and CGS12066B, a 5HT1B receptor subtype selective agonist, which were able to compete for 15-20% of [3H]DHA binding in the nanomolar concentration range, whereas drugs that are selective for other serotonin receptor subtypes inhibited [3H]DHA binding only at much higher concentrations. Another beta-adrenergic receptor antagonist radioligand, [3H]CGP 12177, was found to be more selective for beta-adrenergic receptors. Alprenolol competition curves for [3H]CGP-12177 binding were monophasic and saturation curves, with nonspecific binding defined either by 10 microM alprenolol or by LIGAND, yielded Bmax values close to those obtained with [3H]DHA when its nonspecific binding was defined by LIGAND. [3H]DHA cannot be considered a suitable radioligand to quantify central nervous system beta-adrenergic receptors in the manner in which it has been typically used. PMID- 2546051 TI - Reevaluation of the regulation of beta-adrenergic receptor binding by desipramine treatment. AB - Treatment of rats with desipramine (DMI) has been shown to down-regulate beta adrenergic receptor-stimulated adenylate cyclase and reduce the Bmax of beta adrenergic receptors in some brain areas. Recent reports have indicated that the down-regulation in the number of beta-adrenergic receptors following DMI treatment does not occur if the serotonin system has been impaired following parachlorophenylalanine (PCPA) or 5,7-dihydroxytryptamine injection. We have previously shown that [3H]dihydroalprenolol ([3H]DHA), the most commonly used radioligand to measure central nervous system beta-adrenergic receptors, labels another site under normal experimental procedures, in addition to the beta adrenergic receptors. This second site has some pharmacological characteristics of the 5-hydroxytryptamine1A receptor. The depletion of serotonin following PCPA injection was indeed able to prevent the down-regulation of [3H]DHA binding sites after DMI injection. However, PCPA alone increased the density of [3H]DHA binding sites. If the nonlinear, least squares, curve-fitting program LIGAND was allowed to define [3H]DHA nonspecific binding or if the more selective beta-adrenergic receptor radioligand [3H]CGP-1277 was used, the Bmax of beta-adrenergic receptors was not changed after PCPA injection. Importantly, PCPA did not prevent beta adrenergic receptor down-regulation following DMI treatment. The blockade of 5 hydroxytryptamine2 receptors, via ketanserin administration, during DMI treatment did not change the response of beta-adrenergic receptors. Furthermore, if LIGAND was used to define the nonspecific binding of [3H]DHA, the down-regulation of beta-adrenergic receptors was significant 24 hr after a single DMI injection. The same rapid down-regulation was demonstrated with [3H]CGP-12177. However, if [3H]DHA was used to label beta-adrenergic receptors in the "typical" manner (nonspecific binding defined by 10 microM alprenolol), a decrease in the number of beta-adrenergic receptors was significant only after seven daily DMI injections. These data demonstrate that the use of [3H]DHA to measure beta adrenergic receptors can be misleading, because changes in its second binding site can conceal the changes occurring in beta-adrenergic receptors. Moreover, these results suggest that a similarity in the time course of action of DMI cannot be used to support the hypothesis that its therapeutic antidepressant action is related to beta-adrenergic receptor down-regulation. PMID- 2546052 TI - Calcium-dependent effects of maitotoxin on phosphoinositide breakdown and on cyclic AMP accumulation in PC12 and NCB-20 cells. AB - The marine dinoflagellate toxin maitotoxin (MTX) stimulates phosphoinositide breakdown in pheochromocytoma PC12 cells and in neuroblastoma hybrid NCB-20 cells. In both cell lines, the stimulation of phosphoinositide breakdown by MTX is dependent on extracellular calcium, but it is not reduced by organic or inorganic calcium channel blockers. In PC12 cells, the maximal stimulation of phosphoinositide breakdown occurs at 1.5 mM [Ca2+]o, whereas in NCB-20 cells the maximal stimulation is observed at 2.5-4.5 mM [Ca2+]o. Phosphoinositide breakdown is known to lead to formation of both inositol phosphates and diacylglycerols. The latter, through stimulation of protein kinase C, would, like phorbol esters, be expected to augment cyclic AMP accumulation in PC12 cells and to inhibit receptor-mediated cyclic AMP accumulation in NCB-20 cells. MTX does potentiate forskolin-induced accumulation of cyclic AMP in PC12 cells and does inhibit prostaglandin E2-induced accumulation of cyclic AMP in NCB-20 cells. The effects of MTX on accumulation of cyclic AMP are calcium dependent and the concentrations of calcium required for maximal responses are the same as the ones required for maximal stimulation of phosphoinositide breakdown. MTX increases intracellular calcium in both cell lines, as measured by calcium-quin2 fluorescence. But the effects of MTX on forskolin- and prostaglandin E2-mediated cyclic AMP accumulation are not mimicked by a calcium ionophore and are not blocked by nifedipine, a calcium channel blocker. Translocation of protein kinase C occurs after treatment with MTX in both cell lines; the protein kinase C activity and content are reduced in the cytosol and increased in membranes after exposure to either MTX or a phorbol ester. The results confirm previous studies on the heterogeneous input of protein kinase C to cyclic AMP-generating systems performed with phorbol esters and demonstrate the utility of MTX as a unique tool for studies of systems that involve second messengers generated through stimulation of phosphoinositide breakdown. PMID- 2546053 TI - Podophyllotoxin analogs: effects on DNA topoisomerase II, tubulin polymerization, human tumor KB cells, and their VP-16-resistant variants. AB - Several derivatives of podophyllotoxin with modifications at the C-4 position of ring C, in addition to demethylation at the C-4' position of ring E, were examined for inhibitory activity against DNA topoisomerase II and tubulin polymerization, generation of protein-linked DNA breaks, and cytotoxicity against KB cells and VP-16-resistant KB variants. Substitution of podophyllotoxin with a group in the beta configuration at the C-4 position of ring C resulted in compounds with greater inhibitory activity against DNA topoisomerase II and lower inhibitory activity against tubulin polymerization than those with an alpha configuration. These active analogs exhibited the same mechanism of DNA topoisomerase II inhibition as the epipodophyllotoxin derivative VP-16, which causes protein-linked DNA breaks in vitro as well as in cells. Two analogs selectively inhibited DNA topoisomerases II to a greater extent than tubulin polymerization. These analogs were cytotoxic towards KB cells in addition to VP 16-resistant KB cell lines, which indicated limited cross-resistance with VP-16 in VP-16-resistant KB variants. PMID- 2546054 TI - Purification and properties of the Rous sarcoma virus internal enhancer binding factor. AB - The internal enhancer binding factor (IBF) that specifically binds sequences within the gag gene internal enhancer of Rous sarcoma virus Schmidt-Ruppin A was purified to near homogeneity from BHK cells. The polypeptides that constituted IBF DNA-binding activity were identified by sodium dodecyl sulfate-polyacrylamide gel analysis. As isolated from BHK cells, IBF consisted of two different but related polypeptides. One (IBF alpha) had a molecular weight of 40,000; the other (IBF beta) had a molecular weight of 20,000 and appeared to be a proteolytic product of IBF alpha. The site within the gag gene to which IBF bounds in vitro (internal enhancer site 2; nucleotides 856 to 878 of the Rous sarcoma virus genome) were demonstrated to function as a cis-acting transcriptional stimulatory element both in vivo and in vitro. By using HeLa cell nuclear transcription extracts, purified IBF was found to function as a trans-acting transcription factor that stimulated transcription in vitro. Purified IBF was also demonstrated to be very similar to EBP20 (K. Carlberg, T. A. Ryden, and K. Beemon, J. Virol. 62:1617-1624, 1988), and it may well belong to the same family of DNA-binding proteins. PMID- 2546056 TI - Hygromycin resistance as a selectable marker in Dictyostelium discoideum. AB - We have constructed an expression cartridge which has the bacterial hygromycin resistance gene (hph) fused to the Dictyostelium discoideum actin 15 promoter, with a segment of 3'-flanking DNA from the actin 15 locus placed downstream of the hph gene to serve as a transcription terminator. The plasmid pDE109, which contained this cartridge and a Dictyostelium origin of replication, transformed D. discoideum with high efficiency under hygromycin selection. The availability of this selectable marker circumvents the previous limitation of having G418 resistance as the only selectable marker for this organism; secondary transformation can now be used to introduce DNA into previously transformed cell lines. PMID- 2546055 TI - Inverse regulation of the yeast COX5 genes by oxygen and heme. AB - The COX5a and COX5b genes encode divergent forms of yeast cytochrome c oxidase subunit V. Although the polypeptide products of the two genes are functionally interchangeable, it is the Va subunit that is normally found in preparations of yeast mitochondria and cytochrome c oxidase. We show here that the predominance of subunit Va stems in part from the differential response of the two genes to the presence of molecular oxygen. Our results indicate that during aerobic growth, COX5a levels were high, while COX5b levels were low. Anaerobically, the pattern was reversed; COX5a levels dropped sevenfold, while those of COX5b were elevated sevenfold. Oxygen appeared to act at the level of transcription through heme, since the addition of heme restored an aerobic pattern of transcription to anaerobically grown cells and the effect of anaerobiosis on COX5 transcription was reproduced in strains containing a mutation in the heme-biosynthetic pathway (hem1). In conjunction with the oxygen-heme response, we determined that the product of the ROX1 gene, a trans-acting regulator of several yeast genes controlled by oxygen, is also involved in COX5 expression. These results, as well as our observation that COX5b expression varied significantly in certain yeast strains, indicate that the COX5 genes undergo a complex pattern of regulation. This regulation, especially the increase in COX5b levels anaerobically, may reflect an attempt to modulate the activity of a key respiratory enzyme in response to varying environmental conditions. The results presented here, as well as those from other laboratories, suggest that the induction or derepression of certain metabolic enzymes during anaerobiosis may be a common and important physiological response in yeast cells. PMID- 2546057 TI - Factor interactions with the simian virus 40 early pre-mRNA influence branch site selection and alternative splicing. AB - To study the interaction of splicing factors with the simian virus 40 early region pre-RNA, which can be alternatively spliced to produce large T and small t mRNAs, we used an in vitro RNase protection assay that defines the 5' boundaries of factor-RNA interactions. Protection products reflecting factor interactions with the large T and small t 5' splice sites and with the multiple lariat branch site region were characterized. All protection products were detected very early in the splicing reaction, before the appearance of spliced RNAs. However, protection of the large T 5' splice site was detected well before small t 5' splice site and branch site protection products, which appeared simultaneously. Oligonucleotide-targeted degradation of small nuclear RNAs (snRNAs) revealed that protection of the branch site region, which occurred at multiple sites, required intact U2 snRNA and was enhanced by U1 snRNA, while protection of the large T and small t 5' splice sites required both U1 and U2 snRNAs. Analysis of several pre RNAs containing mutations in the branch site region suggests that factor interactions involving the multiple copies of the branch site consensus determine the selection of branch points, which is an important factor in the selection of alternative splicing pathways. PMID- 2546058 TI - Expression of alpha- and beta-tubulin genes during dimorphic-phase transitions of Histoplasma capsulatum. AB - Recent investigations have confirmed the presence of one alpha-tubulin gene (TUB1) and one beta-tubulin gene (TUB2) in the dimorphic fungus Histoplasma capsulatum. In the present study, Northern blot (RNA blot) analyses revealed multiple alpha-tubulin transcripts and a single beta-tubulin transcript in the yeast and mycelial phases of the high-virulence 217B strain and low-virulence Downs strain. S1 nuclease protection assays demonstrated one initiation start site and two major stop sites for the TUB1 transcripts, suggesting that variations in 3' processing generate the alpha-tubulin messages of 2.5 and 2.0 kilobases. Dot blot hybridization experiments indicated that tubulin gene expression is developmentally regulated during the dimorphic phase transitions. alpha- and beta-tubulin mRNAs increased six- to eightfold during the yeast-to mycelium conversion and decreased two- to threefold during the reverse transition. These changes in tubulin mRNA content coincided with major morphological events associated with H. capsulatum development. Western blots (immunoblots) of H. capsulatum yeast-specific proteins resolved by two dimensional gel electrophoresis demonstrated a single alpha- and a single beta tubulin isoform. Multiple tubulin polypeptides expressed in mycelia are probably products of posttranslational modifications. PMID- 2546059 TI - Localization of transcriptional regulatory elements and nuclear factor binding sites in mouse ribosomal protein gene rpL32. AB - The DNA sequences required for expression of the ribosomal protein gene rpL32 were identified by transient-expression assays of chimeric rpL32-chloramphenicol acetyltransferase genes. These studies showed that maximal rpL32 expression requires sequences in a 150- to 200-base-pair region spanning the transcriptional start site. Three discrete regions of importance were identified: one between positions -79 and -69 and two others located downstream of the transcriptional start site. Progressive 5' or 3' deletions caused stepwise decreases in expression, which suggested a complex interplay of redundant or compensatory elements. Gel mobility shift assays were used to identify trans-acting nuclear factors which bind to segments of the rpL32 promoter that are known to be important for transcription. Evidence for several distinct nuclear factors is presented. The binding sites for these factors were localized to the following regions: -79 to -69, -36 to -19, -19 to +11, +11 to +46 in exon I, and within the first 31 base pairs of intron 1. One of these factors may bind to multiple sites within the promoter region. Interestingly, the factor that binds to a sequence motif in the first exon also binds to similar motifs in a comparable region of the c-myc gene. PMID- 2546060 TI - Glucose-regulated protein (GRP94 and GRP78) genes share common regulatory domains and are coordinately regulated by common trans-acting factors. AB - We isolated the promoter of the human gene encoding the 94,000-dalton glucose regulated protein (GRP94). The 5'-flanking region important for its expression was identified by deletion analysis. Comparison of the promoters of the genes for GRP78 and GRP94 derived from human, rat, and chicken cells revealed a common domain of 28 base pairs within the putative regulatory regions of both genes. This domain has been shown to interact with protein factors in the promoter of the gene for GRP78. Since the genes for GRP94 and GRP78 are transcriptionally regulated with similar kinetics under a variety of stress conditions, we are interested in examining the possible mechanisms for their coordinated expression. Through in vitro and in vivo competition assays, we found that the protein factors which interact with the promoter of the gene for GRP94 also have affinity for the conserved domain of the promoter of the gene for GRP78. These findings suggest that the genes for GRP94 and GRP78 are coordinately regulated through common trans-acting factors which recognize a common regulatory domain of glucose regulated protein gene promoters. PMID- 2546061 TI - Differentiation in vitro of a leukemia virus-induced B-cell lymphoma into macrophages. AB - Cells of the hemopoietic system arise by proliferation and differentiation of progenitor cells. This process begins with multipotential stem cells which can self-renew and also undergo progressive differentiation to progenitor cells committed to particular lineages, ultimately yielding mature blood cells (D. Metcalf and M. A. S. Moore, Haematopoietic Cells, 1971). Early commitment of lymphoid progenitors is generally believed to separate the lymphoid lineage from the myeloid and erythroid lineages, whose progenitors are separated late in differentiation (Metcalf and Moore, 1971). We recently developed a derivative of Moloney murine leukemia virus (M-MuLV) in which the enhancer sequences from simian virus 40 were substituted into the M-MuLV long terminal repeat. This recombinant virus (delta Mo + SV M-MuLV) induces pre-B and B lymphoid leukemia with long latency after inoculation of 2-day-old NIH Swiss mice (R. Hanecak, P. K. Pattengale, and H. Fan, J. Virol. 62:2427-2436, 1988). In this report, we describe the derivation of a permanent, virus-producing cell line with the phenotypic characteristics of mature macrophages from a B-cell-derived lymphoblastic lymphoma induced by delta Mo + SV M-MuLV. Comparison studies of immunoglobulin heavy-chain gene rearrangements and also delta Mo + SV M-MuLV proviral integration sites confirmed that the macrophage cell line was derived from the original B-lymphoblastic lymphoma. Moreover, inoculation of the macrophage cell line into animals resulted in histiocytic sarcomas of the macrophage type, thus reflecting stable conversion of B-lymphoid tumor cells to the macrophage phenotype. These results suggest a closer relationship between lymphoid and myeloid cells than previously believed. PMID- 2546062 TI - Epidermal growth factor induction of cellular proliferation and protooncogene expression in growth-arrested rat H4IIE hepatoma cells: role of cyclic adenosine monophosphate. AB - The precise molecular events involved in growth factor-mediated cell proliferation in eukaryotes have not been entirely elucidated. Identification and characterization of the itnracellular molecular signaling systems linking growth factor function with nuclear events would provide insight into the regulatory mechanisms governing eukaryotic cell growth. In this report, we demonstrate that serum-deprived rat H4IIE hepatoma cells enter a quiescent state and remain viable in the absence of serum for up to 7 days. These cells can be stimulated to transverse the cell cycle and proliferate in response to epidermal growth factor (EGF) after a 24-h lag phase. We were able to completely mimic the mitogenic effects of EGF with 8-p-chlorophenylthio-cAMP (8-CPT-cAMP) but only partially with N6-(Bu)2-cAMP. EGF and 8-CPT-cAMP together induce a synergistic increase in H4IIE hepatoma cell proliferation. The calcium ionophore A23187 and the phorbol ester, 4 beta-phorbol 12-myristate 13-acetate had little effect on H4IIE cell proliferation. EGF treatment led to a rapid and transient increase of intracellular cAMP concentration. Both 8-CPT-cAMP and EGF were also equally effective in causing a rapid and transient induction of c-fos and c-myc protooncogene mRNA levels when added to growth-arrested H4IIE cells while A23187, N-(Bu)2-cAMP, and 4 beta-phorbol 12-myristate 13-acetate were significantly less effective. Both EGF and 8-CPT-cAMP affect protooncogene expression in growth arrested rat H4IIE hepatoma cells primarily at the transcriptional level. Localization and semi-quantification of nuclear pp55c-fos and 63 (kilodalton)-myc protooncoproteins by immunocolloidal gold electron microscopy revealed that EGF and/or 8-CPT-cAMP treatment of quiescent H4IIE hepatoma cells led to a marked and rapid nuclear accumulation of these proteins in discrete nuclear substructures. Cummulatively, these results suggest that cAMP participates in the intracellular signaling system mediating the mitogenic and protooncogene inducing effects of EGF on growth-arrested rat H4IIE hepatoma cells. PMID- 2546063 TI - Regulation of the cellular retinoid-binding proteins and their messenger ribonucleic acids during P19 embryonal carcinoma cell differentiation induced by retinoic acid. AB - P19 embryonal carcinoma (EC) cells can be induced to differentiate in vitro into a variety of cell types by treatment with different concentrations of retinoic acid (RA). A study was conducted to explore the regulation of expression of the genes for cellular retinoic acid-binding protein (CRABP) and cellular retinol binding protein (CRBP) in P19 cells induced to differentiate by RA. For each retinoid-binding protein, both the level of specific mRNA and of immunoreactive protein were measured, respectively, by RNase protection assay and by a specific RIA. Dramatic increases in CRABP and CRBP were seen, at both the mRNA and protein levels, during the RA-induced differentiation. CRBP induction differed from that of CRABP in several major ways. 1) Induction of CRBP occurred at lower concentrations of RA (10(-9) M) than did that of CRABP (10(-8)-10(-7) M). 2) CRBP induction was an early response (within 3 h) to RA treatment, whereas CRABP induction occurred at a later time (12-24 h). 3) Induction of CRABP mRNA by RA was blocked by the protein synthesis inhibitor cycloheximide, whereas induction of CRBP mRNA was not. 4) Several differentiation inducers were tested for their effects on the expression of CRABP and CRBP in P19 cells. CRBP induction occurred with a wider spectrum of inducers than did that of CRABP. 5) In addition, the induction of CRABP and CRBP mRNAs by RA was examined in six different cell lines, including three EC lines. CRBP induction occurred in a wider spectrum of cell lines than did that of CRABP. The induction of CRABP in EC cells seems, in general, to correlate with their differentiation into neuron-like cells. Taken together, our results suggest that CRBP induction may be a direct response to RA and represent a general event in RA-induced cell differentiation, whereas CRABP induction may be an indirect response and represent a later event restricted to only certain differentiation pathways. CRBP may be an early response gene induced by RA. PMID- 2546064 TI - Leucine zipper in human DNA topoisomerase II. AB - Examination of the amino acid sequence of human DNA topoisomerase II revealed the presence of a leucine zipper, a novel motif found in several proteins localized to the cell nucleus. The presence of this motif in this unique protein may explain some of the normal functions of topoisomerase II as well as the disruption of those functions by antineoplastic drugs. PMID- 2546065 TI - [Granulomatous hepatitis in cytomegalovirus infection]. AB - Authors report the serologically verified CMV hepatitis of a 35-year old man whose hepatic alteration appeared in the form of a granulomatous hepatitis. On this account the characteristics of granulomatous hepatitis cases published so far in the literature are surveyed. PMID- 2546066 TI - Giant-cell neoplasm of the sphenoid sinus. AB - A rapidly growing giant-cell neoplasm arising in the sphenoid sinus of a 26-year old man is reported. The patient's only initial symptoms were severe headaches and left sixth-nerve palsy. Computerized tomography of the brain showed extensive tumor infiltration of the bony structures and soft tissues of the base of the skull. A transsphenoidal surgical procedure proved unsuccessful in removing the tumor. Histologic examination revealed a proliferation of osteoclast-type giant cells against a background of highly atypical mononuclear cells, raising the question of malignancy. The differential diagnosis and problems in nomenclature of this lesion are discussed. PMID- 2546067 TI - Mesothelial papillary proliferation of the pleura associated with radiation therapy: does it have a role in the pathogenesis of mesothelioma? AB - Diffuse papillary proliferation of mesothelial cells in the pleura mimicking metastatic carcinoma was seen four weeks following radiation therapy for a Pancoast tumor. Such papillary proliferations are not observed incidentally and are envisioned to occur during asbestos-induced carcinogenesis. We postulate that similar papillary lesions may serve as a link in the pathogenesis of radiation induced mesotheliomas. PMID- 2546068 TI - [The role of chemotaxis genes in establishing the associative relations between Azospirillum brasilense and wheat]. AB - The chemotactic properties of a number of Azospirillum brasilense natural strains have been studied. Azospirillum demonstrate the positive chemotactic reaction towards the organic acids salts but a poor reaction towards the presence of the attractants like hydrocarbons and aminoacids except for arabinose and glutamic acid. The series of Che- mutants deficient in general chemotaxis has been selected by introducing the transposon Tn5 into the cells of rifampicinresistant mutant strain Azospirillum brasilense 5T-2. The ability of the mutant cells to fast and solid adsorption on the roots of the sterile wheat sprouts is shown to be decreased 2-3 folds as compared with the one of the wild type strain. Chemotaxis is suggested to affect the adsorbtion ability of azospirillum and their associative properties. PMID- 2546069 TI - [Possible role of translation initiation factors in the regulation of expression of attenuated mutations of poliovirus]. AB - Initiation of poliovirus RNA translation in reticulocyte lysates is mainly not precise, i.e. it occurs at the sites in the middle of the viral genome but not at the beginning of the polyprotein reading frame. The anomaly is due to the deficiency of translation initiation factors. Partial purification of the protein fraction stimulating the precise translation from the Krebs-2 cells is reported in the paper. This fraction, like the crude lysates factors, was considerably less active with the RNA of attenuated poliovirus strains of type 1 and 3 than with the RNA of virulent strains. The change in interaction of the specific segment of viral RNA with the translation initiation factors is suggested to contribute to the attenuated phenotype of the Sabin poliovirus strains. PMID- 2546070 TI - [Current approaches to the search for new restriction endonucleases]. AB - The main methodological approaches to the search of new restriction endonucleases are reviewed. These methods include obtaining acellular extracts by ultrasonic desintegration of microbial cells, osmotic shock effects, the effects of organic solvents, mechanical disruption of bacterial cells, biphase division after Albertson and others. The resolving power of any method discussed depends mainly on the level of restriction endonuclease activity, the presence of nonspecific endonucleases in the biomass, the presence of exonucleases and the taxonomy of the used microorganisms. PMID- 2546072 TI - DNA amplification in genetic toxicology. AB - Chemical compounds can cause amplification of specific DNA sequences. DNA amplification may result in an enhanced production of gene products which help cells to cope with the chemicals. This may lead to a resistance of the cells toward the agent. Additionally, initiation of transformation or progression of transformed cells to tumorigenicity may also involve DNA amplification. Therefore, it is of interest to study the potential of chemicals to induce DNA amplification. This report focuses on the investigation of a variety of chemicals in 2 systems with which the amplification of viral DNA is measured within cells in culture. One model system comprises the measurement of SV40 DNA content in an SV40-transformed Chinese hamster cell line following chemical treatment. Antitumor agents as well as genotoxic and non-genotoxic compounds were studied in this system as a first step to determine the DNA amplification-inducing potential of a variety of differently acting chemical compounds. Also, a novel assay based on adeno-associated virus infection of cells is described. This system may offer the possibility of studying DNA amplification in a variety of different target cells. For the future, the need is stressed to develop and analyze versatile systems to study amplification of specific target genes in untransformed cells and in tumor cells. PMID- 2546071 TI - [Electrophoretic analysis of genome RNA of human rotaviruses]. AB - The schemes for profiles of migration of RNA segments from human rotaviruses isolated in Gorky city in 1981-1987 are presented. The groups of the "long", "short" and "wide" electrophoretypes were identified. The dominant rotaviral phoretypes are determined. The possibilities of electrophoretyping and prospects of the future research are discussed. PMID- 2546073 TI - Restriction endonucleases do not induce sister-chromatid exchanges in Chinese hamster ovary cells. AB - Bacterial restriction enzymes offer the unique opportunity to determine the biological and cytogenetic consequences of DNA double-strand breakage. To examine the role of various types of breaks in sister-chromatid exchange (SCE) formation, we used restriction enzymes with different recognition sequences and different cutting frequencies to generate DNA double-strand breaks in Chinese hamster ovary (CHO) cells. The restriction enzymes were introduced by electroporation into exponentially growing cells during the second replication cycle in bromodeoxyuridine, and SCEs were analyzed at mitosis. Contrary to results reported by others, we found no increase in SCE frequency in cells exposed to restriction enzymes despite the presence of numerous cells with chromatid aberrations. These data suggest that DNA double-strand breaks do not lead to SCE formation. PMID- 2546074 TI - AAEE case report #17: Peripheral neuropathy in monoclonal gammopathy of undetermined significance. AB - Monoclonal gammopathy of undetermined significance (MGUS) is the most common paraproteinemia associated with polyneuropathy. Although the clinical and electrodiagnostic manifestations most resemble those of chronic inflammatory demyelinating polyneuropathy, some patients manifest a pure sensory neuropathy or neuronopathy. The M protein is usually IgM, and its concentration in serum is low. Nerve pathology from patients with demyelinating disease shows a reduction of large myelinated fibers and segmental demyelination with remyelination. In some cases, the M protein possesses antibody activity against components of the myelin sheath or axon. These neuropathies may respond to treatment with steroids, immunosuppressant agents, and plasma exchange. PMID- 2546076 TI - Myasthenic U-shaped decrement in multifocal cervical radiculopathy. AB - A middle-aged man with lymphomatous meningitis developed acute cervical radiculopathy. Repetitive stimulation of an affected nerve revealed a "U-Shaped" decrement. The decrement was absent 10 days later. We hypothesize that the decrement was secondary to simultaneous denervation of many motor units and disappeared as denervation became complete in those motor units. PMID- 2546075 TI - Prediction by 31P-NMR of the irreversibility of ischemic injury in rat skeletal muscle after ligation of the femoral artery. AB - Rat leg muscles, rendered ischaemic 1 hour previously by ligation of the femoral artery, were submitted to 20 minutes exercise by electrical stimulation of the sciatic nerve. 31P-NMR spectroscopy was used to monitor the changes in high energy phosphate content of the muscles before, during, and after exercise. Fifteen of the 35 studied muscles evolved toward total necrosis, whereas the others showed signs of recovery over a 2-5-hour postexercise period. Those muscles which did not subsequently recover contained significantly more inorganic phosphate (Pi) at rest (before exercise) than those which recovered. It is suggested that under acute ischaemic conditions the Pi level at rest is correlated with the extent of blood flow restriction and can be used to predict the severity of the ischemia. PMID- 2546077 TI - Molecular targets of antiviral therapy. PMID- 2546078 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 29-1989. A 79-year-old man with fever, abdominal pain, and an inflamed right eye. PMID- 2546080 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 32-1989. A 61-year-old man with multiple pulmonary nodules, solid and cavitary. PMID- 2546079 TI - The risk of breast cancer after estrogen and estrogen-progestin replacement. AB - To examine the risk of breast cancer after noncontraceptive treatment with estrogen, we conducted a prospective study of 23,244 women 35 years of age or older who had had estrogen prescriptions filled in the Uppsala region of Sweden. During the follow-up period (mean, 5.7 years) breast cancer developed in 253 women. Compared with other women in the same region, the women in the estrogen cohort had an overall relative risk of breast cancer of 1.1 (95 percent confidence interval, 1.0 to 1.3). The relative risk increased with the duration of estrogen treatment (P = 0.002), reaching 1.7 after nine years (95 percent confidence interval, 1.1 to 2.7). Estradiol (used in 56 percent of the treatment periods in the cohort) was associated with a 1.8-fold increase in risk after more than six years of treatment (95 percent confidence interval, 0.7 to 4.6). No increase in risk was found after the use of conjugated estrogens (used in 22 percent of the treatment periods) or other types, mainly estriols (used in 22 percent of the treatment periods). Although the numbers of women were smaller, the risk of breast cancer was highest among the women who took estrogen and progestin in combination for extended periods. The relative risk was 4.4 (95 percent confidence interval, 0.9 to 22.4) in women who used only this combination for more than six years. Among women who had previously used estrogens alone, the relative risk after three years or more of use of the combination regimen was 2.3 (95 percent confidence interval, 0.7 to 7.8). We conclude that in this cohort, long-term perimenopausal treatment with estrogens (or at least estradiol compounds) seems to be associated with a slightly increased risk of breast cancer, which is not prevented and may even be increased by the addition of progestins. PMID- 2546082 TI - Carcinogenesis. Is imprinting to blame? PMID- 2546081 TI - Treatment of disseminated histoplasmosis in hamsters. AB - A comparative study between itraconazole, ketoconazole and amphotericin B in the treatment of experimental histoplasmosis in hamsters was carried out. Seventy five animals were inoculated intracardiacally with the yeast-phase of Histoplasma capsulatum. They were divided in 5 groups: 1) treated with itraconazole by gavage (g) at a daily dose of 16 mg/kg; 2) treated with ketoconazole by (g) at a daily dose of 80 mg/kg; 3) treated with amphotericin B intraperitoneally (i.p.) at 6 mg/kg every other day; 4) control animals receiving distilled water i.p. and 5) control animals receiving P.E.G. 200 by (g). All the treatments were started one week after the challenge inoculation and they were given for 21 days. The results were evaluated by autopsy of all the animals one week after the end of the treatments. The following determinations were taken into account: microscopic examinations of spleen, liver and lungs and cultures of the spleen with determination of colony forming units/g. All the antifungal drugs used in this study were able to cause negative microscopic examinations of the liver, spleen and lungs; but only amphotericin B produced culture negative results. Itraconazole and ketoconazole presented 66% and 86% of positive cultures respectively, nevertheless the C.F.U. were lower than those obtained in control groups. In these experimental conditions amphotericin B seems to be more active than the azolic compounds and itraconazole is slightly superior to ketoconazole at a lower dose. PMID- 2546083 TI - Palindromic sequences in heteroduplex DNA inhibit mismatch repair in yeast. AB - Although single heterozygous markers in yeast usually segregate during meiosis in a 2:2 ratio, abberant 3:1 segregations occur quite frequently as a result of gene conversion events. A second type of aberrant segregation, post-meiotic segregation, results from the segregation of two genotypes from a single haploid spore; in yeast such events are detected as sectored spore colonies and usually occur rarely. Post-meiotic segregation is thought to result from the replication of heteroduplex DNA formed during meiotic recombination. We report here that if the heteroduplex includes a palindromic insertion sequence, a high frequency of post-meiotic segregation results. This suggests that palindromic insertions are poorly repaired, which may be the result of hairpin-loop formation that affects the efficiency of repair of heteroduplex DNA. PMID- 2546084 TI - Retro-secretion of recombinant proteins. AB - The retroviral Gag protein can be used to package recombinant proteins produced by mammalian cell cultures. Protein products can then be isolated easily from the recombinant virions. PMID- 2546087 TI - Involvement of opioid and dopaminergic systems in isolation-induced pinning and social grooming of young rats. AB - Pinning, as a measure for play, and social grooming were simultaneously studied in juvenile rats. Short-term social isolation increased both behavioural responses. This increase was attenuated by the opioid antagonist naltrexone, whilst the opiate, morphine, and the opioid peptide beta-endorphin, increased the responses. Pinning was more sensitive to the effects of naltrexone, whilst beta endorphin stimulated particularly social grooming. Small doses of the dopaminergic drug, apomorphine, decreased both pinning and grooming behaviour of the short-term isolated rats. Some of the effects were partially antagonized by the dopamine antagonist haloperidol, and the neurolepticum-like peptide, desenkephalin-gamma-endorphin (DE-gamma-E). A small dose of haloperidol and DE gamma-E stimulated social grooming in particular, whilst a larger dose of haloperidol decreased pinning and social grooming. It is concluded that both opioid and dopaminergic systems are implicated in the increase of pinning and social grooming induced by short-term social isolation. The differences in sensitivity of pinning and social grooming for opioid and dopaminergic drugs and peptides are discussed in relation to possible differences in the neural systems underlying both social activities. PMID- 2546086 TI - Failure of non-adrenergic transmission in the rat vas deferens. AB - Repetitive field stimulation of the vas deferens of the rat with long trains of pulses (90 pulses, 10 Hz, 1 msec) at intervals of 30 or 60 sec produced a decline in the heights of contractile responses. This decline in responses was much more pronounced in phentolamine (5 microM)-treated than in untreated preparations and in the prostatic rather than in the epididymal end of the vas deferens. The decline in the responses was dependent on the number of pulses/train, the frequency of pulses in the train and the interval between successive trains. Potassium chloride (KCl)-induced and adenosine triphosphate (ATP)-induced contractions were largely unaltered after 30 min of stimulation. The decline in the responses was significantly reduced in low but not in high extracellular calcium. 4-Aminopyridine (4-AP) caused an initial potentiation of contractile responses, followed by a faster decline of responses. The decline in responses was more intense in vas deferens from reserpine-treated animals. The results support the view that electrical stimulation-induced decay of motor transmission in the vas deferens of the rat is caused by a specific failure of the non adrenergic transmission. PMID- 2546085 TI - Na+, K+-ATPase activity in cultured C6 glioma cells. AB - Rat C6 glioma cells were cultured for 4 days in MEM medium supplemented with 10% bovine serum and Na+, K+-ATPase activity was determined in homogenates of harvested cells. Approximately 50% of enzyme activity was attained at 1.5 mM K+ and the maximum (2.76 +/- 0.13 mumol Pi/h/mg protein) at 5 mM K+. The specific activity of Na+, K+-ATPase was not influenced by freezing the homogenates or cell suspensions before the enzyme assay. Ten minutes' exposure of glioma cells to 10( 4) or 10(-5) M noradrenaline (NA) remained without any effect on NA+, K+-ATPase activity. Neither did the presence of NA in the incubation medium, during the enzyme assay, influence the enzyme activity. The nonresponsiveness of Na+, K+ ATPase of C6 glioma cells to NA is consistent with the assumption that alpha (+) form of the enzyme may be preferentially sensitive to noradrenaline. Na+, K+ ATPase was inhibited in a dose-dependent manner by vanadate and 50% inhibition was achieved at 2 x 10(-7) M concentration. In spite of the fact that Na+, K+ ATPase of glioma cells was not responsive to NA, the latter could at least partially reverse vanadate-induced inhibition of the enzyme. Although the present results concern transformed glial cells, they suggest the possibility that inhibition of glial Na+, K+-ATPase may contribute to the previously reported inhibition by vanadate of Na+, K+-ATPase of the whole brain tissue. PMID- 2546088 TI - Gostatin blocks physiological actions and binding of acidic amino acids in rat brain. AB - Gostatin, an inhibitor of aspartate aminotransferases isolated from Streptomyces sumanensis NK-23, was tested as an antagonist of acidic amino acid-mediated responses in the in vitro hippocampal slice and of acidic amino acid binding. Gostatin blocked responses to N-methyl-aspartate (AMPA) or L-glutamate. Gostatin also displaced N-methyl-D-aspartate-sensitive [3H]-L-glutamate binding (Ki = 22.0 microM) more potently than [3H]-kainate binding. Gostatin appears to be a relatively nonselective acidic amino acid antagonist in the mammalian central nervous system. PMID- 2546089 TI - MU and delta opioid receptors control differently the horizontal and vertical components of locomotor activity in mice. AB - The present study investigated the role of mu and delta opioid receptors in the control of the horizontal and vertical components of locomotion. Mice received intracerebroventricularly (i.c.v.) enkephalin analogs specific for either the mu or delta opioid receptors. The administration of the specific mu agonist [D-Ala2 NMePhe4-Gly5(ol)] enkephalin (DAGO) induced a dose-dependent increase in horizontal activity and a decrease in vertical activity. The specific delta agonist [D-Pen2,D-Pen5] enkephalin (DPDPE) increased both components of motor activity. The opiate antagonist naltrexone reversed the effects of DAGO, but did not influence the effects of DPDPE on motor activity. The pretreatment with the delta opiate antagonist ICI 154, 129 completely reversed the effects of DPDPE on locomotion but antagonized only partially the effects of DAGO on locomotion. These results indicate that the two components of locomotor activity--horizontal and vertical activity--are modulated differently by the stimulation of mu or delta opioid receptors. PMID- 2546090 TI - Omega-conotoxin GVIA specifically blocks neuronal mechanisms in rat ileum. AB - Omega Conotoxin is a peptide venom of the marine snail Conus geographus which has been postulated as a neural specific calcium channel blocker in several systems. In the present study in rat ileum strips omega-conotoxin GVIA specifically blocked neuronal responses evoked by electrical field stimulation in a dose dependent manner, but did not block responses induced by carbachol or direct depolarization with KCl. On the other hand the dyhydropyridine calcium channel blocker nitrendipine (10(-7) M) showed no specificity and blocked electrical field stimulation as well as carbachol- and KCl-induced contractions. This indicates that contractions of intestinal smooth muscle which can be blocked by dihydropyridine calcium channel blockers or Ca2+-free medium are not reduced by the neurotoxin omega-conotoxin. Omega-conotoxin caused even a little increase of the responses elicited by depolarisation with KCL. Omega-conotoxin blocked however neural mediated contractions which might suggest that it interacts with a distinct neuronal calcium channel and, in addition to tetrodotoxin, could provide a useful pharmacological tool to discriminate between muscular and neural sites of action of excitatory or inhibitory agents. PMID- 2546091 TI - Structure-activity studies of neurokinin A. AB - A structure-activity study on neurokinin A and its C-terminal fragment NKA (4-10) has been performed in order to find selective agonists for the NK-2 receptor and identify chemical modifications suitable for protecting the peptides from degradation, while maintaining activity. Five series of compounds have been prepared and tested: 1. the complete series of the L-Ala monosubstituted analogues of NKA; 2. a series of NKA fragments from the C- or N-terminal; 3. the complete series of NKA (4-10) analogues monosubstituted with beta-Ala; 4. a series of NKA (4-10) analogues with monosubstitutions in pos. 4, 8, 10 or multisubstitutions in two or more of the same positions; and 5. a series of 6 NKA (4-10) analogues monosubstituted with 1-amino,1-cyclohexane carboxylic acid residue. It has been found that the most selective agonists for the NK-2 receptor system are [beta Ala8]NKA (4-10) and [Nle10]NKA (4-10). Protection from aminopeptidase may be obtained by acetylation of the N-terminal amide of NKA (4 10), while partial protection from endopeptidases should be expected from the presence of beta-Ala in position 8. Conformational constraints induced with 1,amino,1-cyclohexane carboxylic acid residue gave weakly active compounds. Multiple substitutions reduce rather than potentiating the favorable effects of the corresponding monosubstituted compounds. PMID- 2546092 TI - Insulin and insulin-like growth factor I in brain tumors: binding and in vitro effects. AB - We have measured insulin and insulin-like growth factor I (IGF-I) binding in human gliomas, meningiomas, and normal brain and studied the effect of insulin on the morphology, proliferation, and differentiation of central nervous system tumor and normal fetal cells in culture. Specific 125I-insulin and 125I-IGF-I binding was demonstrated by competition-inhibition binding assays. Insulin binding was measured in plasma membrane preparations from 9 freshly isolated human meningiomas, 4 glioblastomas multiforme (GBMs), a low-grade glioma, a normal adult brain, and a fetal brain. IGF-I binding was measured in similar preparations from 5 meningiomas, 4 GBMs, a low-grade glioma, and a normal adult brain. Incubations were carried out at 4 degrees C for 18 to 20 hours. Meningiomas showed higher specific insulin binding per 0.25 mg of protein than GBMs (19% versus 3%, P less than 0.005), and this difference was not related to small differences observed in insulin degradation. By contrast, IGF-I binding was significantly higher in gliomas than in meningiomas (27% versus 12%, P less than 0.05). Also, IGF-I binding was significantly higher than insulin binding in GBMs (27% versus 3%, P less than 0.03); binding of both IGF and insulin was high in meningiomas. In normal adult brain IGF-I and insulin binding was 7 to 10%. The ability of insulin to support and enhance the growth of central nervous system tumor cells in culture was investigated. Cell cultures were derived from a freshly isolated glioblastoma, a low-grade glioma, and 3 meningiomas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546093 TI - Treatment of recurrent gliomas with 1,3-bis(2-chloroethyl)-1-nitrosourea and alpha-difluoromethylornithine. AB - Thirty-eight patients with primary recurrent anaplastic gliomas and glioblastomas, were treated with 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and the polyamine inhibitor alpha-difluoromethylornithine (DFMO). There were 5 brain stem, 1 cerebellar, and 32 supratentorial glioma tumors. All had been treated with surgery (except in the case of 4 brain stem tumors for which biopsies were not obtained) and radiotherapy. Eight patients had received prior chemotherapy. Of the 21 patients with evaluable supratentorial anaplastic gliomas, 2 (9.5%) had a partial response and 10 (47.6%) had stable disease. The median time to tumor progression for the anaplastic gliomas has not been attained yet. However, median survival for these 12 patients was 119 weeks measured from the initiation of chemotherapy. Median survival for the entire anaplastic glioma group of 21 was 56 weeks. Minimal activity was seen against glioblastoma multiforme. The median time to tumor progression was 8 weeks with median survival of 21 weeks. Of the 5 patients with brain stem tumors, 3 are alive with stable disease at 77, 93, and 220 weeks. The combination was well tolerated with dose-limiting toxicity being myelosuppression and hearing loss. Further trials are warranted to compare the combination of BCNU and DFMO against BCNU alone in a prospective randomized trial. PMID- 2546094 TI - Lateralization of opioid receptors and their putative ligands in the visual cortex of the tortoise. PMID- 2546096 TI - Rare case of a primary malignant fibrous histiocytoma of the brain. AB - A rare case of primary MFH of the brain is described together with the CT and MRI findings. The radiological appearance of this tumor is discussed in conjunction with the material published in other case reports. PMID- 2546095 TI - Neuronal and neurochemical mechanisms of hypothalamic inhibition of the nociceptive reflex. AB - Using cats anesthetized with chloralose-Nembutal we have studied the effect of high frequency stimulation of the medial and lateral structures of the posterior, tuberal, and anterior hypothalamus, and also of the central gray matter of the midbrain on the nociceptive jaw-opening reflex induced by tooth pulp stimulation. We recorded the EMG response of the digastric muscle as the index of the nociceptive reflex. We have shown that the EMG response of the nociceptive reflex is effectively suppressed on stimulation of all hypothalamic structures and the central gray matter, the threshold of the suppressive action being lower on stimulation of the central gray matter. The effects of the hypothalamic suppression of the nociceptive reflex were eliminated almost completely after naloxone administration, with the exception of the central gray matter, which is slightly more resistant to the action of this agent. After bilateral electrolytic destruction of the central gray matter the antinociceptive effect of the hypothalamus was retained, decreasing only insignificantly, The effect of complete suppression of the amplitude of the EMG response, similar to the effect of stimulation of the hypothalamus and the central gray matter, was also observed after intravenous administration of phenapidine, an opiate agonist with a marked central analgesic action. The neuronal and neurochemical mechanisms of hypothalamic suppression of the nociceptive jaw-opening reflex are discussed. PMID- 2546097 TI - Nerve growth factor receptor is associated with cholinergic neurons of the basal forebrain but not the pontomesencephalon. AB - Sequential immunohistochemical demonstration of nerve growth factor receptor and cholinergic acetyltransferase on the same tissue section in the rat revealed that approximately 92% of all cholinergic neurons in the basal forebrain possessed that receptor. Only 0.9% of the neurons demonstrating nerve growth factor receptor in the basal nuclear complex lacked the cholinergic synthetic enzyme, and a similarly small percentage of cholinergic cells, 7.1%, were choline acetyltransferase-positive but nerve growth factor receptor-negative. Affiliation of nerve growth factor receptor with structural entities morphologically indistinguishable from those demonstrating choline acetyltransferase on separate but corresponding tissue sections was also observed in the telencephalic fiber tracts and terminal fields of basal forebrain cholinergic neurons, including cholinergic puncta in the reticular nucleus of the thalamus. Nerve growth factor receptor was not found in association with choline acetyltransferase-positive somata of the pedunculopontine and laterodorsal tegmental nuclei, however, nor were fibers immunoreactive for nerve growth factor receptor observed originating from those cell bodies. These results suggest that nerve growth factor receptor, which is probably synthesized in cholinergic basal forebrain somata and transported throughout their dendritic and axonal arbors, has a physiologic role in those cells in the adult nervous system. This does not appear to be the case for phenotypically similar neurons of the pontomesencephalotegmental cholinergic complex. PMID- 2546098 TI - Pharmacological studies of voltage-sensitive Ca2+-channels involved in the release of vasoactive intestinal peptide evoked by K+ in mouse cerebral cortical slices. AB - Three types of voltage-sensitive Ca2+-channels, denominated T, N and L, have recently been identified in the nervous system. This classification is based on both the electrophysiological and pharmacological properties of each type of channel. The increase in free intracellular Ca2+ concentration that results from the opening of voltage-sensitive Ca2+-channels triggers various cellular processes. One such process is the depolarization-induced release of neurotransmitters. Owing to the rather selective sensitivity of each type of voltage-sensitive Ca2+-channel to certain antagonists, attempts have recently been carried out to determine which of the T, N or L channels mediates neurotransmitter release. In the present study we have applied such a strategy to the study of the release of vasoactive intestinal peptide from mouse cerebral cortical slices. The release of vasoactive intestinal peptide evoked by K+ 20 mM is inhibited in a concentration-dependent manner by Co2+, Ni2+ and Mn2+ while it remains unaffected by diltiazem 20 microM, nifedipine 10 microM, omega-conotoxin 1 microM and Cd2+ up to 100 microM. Such a pharmacological profile indicates that voltage-sensitive Ca2+-channels of the N and L types are not involved in the release of vasoactive intestinal peptide. These observations are in contrast to what has been described for amine release in the central nervous system, which appears to be mediated by voltage-sensitive Ca2+-channels of the N type.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546099 TI - Proximodistal gradients of the postjunctional folds at the frog neuromuscular junction: a scanning electron microscopic study. AB - Frog endplates were studied with the scanning electron microscope following the removal of the presynaptic terminal by collagenase and acid treatments. Endplates had 2-14 branches of primary cleft. The longest branches were parallel to the muscle fiber. Short branches oblique or perpendicular to the muscle fiber were also present near the central region of the endplates. The openings of postjunctional folds in the primary cleft were clearly visible at the bottom of the primary cleft and could be counted and measured. The longest primary cleft branches of each endplate were divided into segments of 20 microns (length corrected for shrinkage). The number of postjunctional folds per micrometer of primary cleft, the average postjunctional fold length (i.e. across the primary cleft) and the total postjunctional fold's length per micrometer of primary cleft were evaluated for each 20-microns segment of primary cleft. Negative proximodistal gradients were observed for these three parameters for the long branches of primary cleft, i.e. values were higher in the proximal region (near the motor axon) than in the distal region. These postsynaptic gradients probably reflect similar or smaller proximodistal presynaptic gradients for the active zones along the nerve. PMID- 2546101 TI - Computer tomographic assessment of perifocal edema surrounding tumors of the cerebral cortex. AB - It is well known, that intracranial tumors can induce peritumoral edema of variable extension. Some authors describe a dependency between malignancy of tumor and size of perifocal edema. It is generally accepted that metastases induce the largest perifocal edema. This study investigates the relationship between the volume of the perifocal edema and its distance from the surface of the cortex. PMID- 2546100 TI - Visualizing receptors for neurotransmitters in the human brain with autoradiography. AB - Receptors for neurotransmitters and drugs are now well characterized at the molecular level. Thanks to the development of numerous radiolabeled molecules and to the use of autoradiography it is possible to study the characteristics and distribution of these receptors with the anatomical resolution of the microscope. We have used quantitative receptor autoradiography to examine receptors in the human brain and to study receptor alterations in neurodegenerative diseases of the human brain. Alterations in the density and distribution of receptors have been found in diseases such as amyothrophic lateral sclerosis, Huntington's Chorea, Parkinson's disease and Alzheimer's disease. In these diseases different types of receptor alterations have been found. The most characteristic ones are selective receptor losses associated with neuronal losses. Alterations such as receptor hypersensitivity due to degeneration of target areas have also been observed. In some cases no correlations between alterations of the neurotransmitters and receptors have been found. These results indicate that different receptors are associated with specific neuronal systems and could be used as markers for these neuronal populations in different pathological studies. The possibility of visualizing receptors in the living human with non-invasive techniques such as PET could lead to the future use of receptor alterations as a diagnostic tool. PMID- 2546103 TI - [Crohn disease and carcinoma of the small intestine]. AB - A case of cancer of the small bowel associated with Crohn's disease with onset 21 years after the first clinical signs of the latter is reported. The patient survived for 20 months after cancer diagnosis. After noting the rare association between the two conditions, the aetiopathogenetic relations that can be called on to explain relations of causality are analysed and it is emphasised that it is impossible to distinguish symptoms from those of cancer due to the renewed inflammatory activity. Finally it is considered that to clarify unequivocably the relations between the two conditions further studies are required. PMID- 2546102 TI - [Calcium antagonists vs ACE inhibitors in the treatment of essential arterial hypertension in the aged]. AB - Numerous epidemiological studies have shown that systolic and systodisystolic hypertension constitute major risk factors for damaging or fatal cardiovascular accidents in the elderly as well as the young. Furthermore reducing the blood pressure also reduces the risk. In 1983 Fleckenstein investigated the Ca++ and MG++ contact of human arteries and clearly demonstrated that titres of both but especially Ca++ in the arterial wall increased progressively with age. The Authors themselves caused calcinosis of the arterial wall in rats treated with Vitamin D3 and Dihydrotachysterol and were able to prevent the occurrence with Verapamil. It is against this background that the present study compared the efficacy and tolerability of two anti-hypertensive drug groups in the calcium antagonists and the ACE inhibitors (Enalapril Maleate) used individually on two groups of elderly hypertensives. A group of 123 out patients with a mean age of 73 and all suffering from slight-to-moderate hypertension were monitored for 6 months being subjected to the following examinations: clinical assessment including blood pressure measurements lying and standing, biohumoral tests, remote heart X-rays, echocardiography (to establish the Reichek systolic wall stress index) and ECG. The clinical examination and ECG were repeated every 2 weeks for the first 6 months and once a month thereafter. The heart X-rays, echocardiogram and biohumeral tests were performed every 6 months. The patients were divided into two groups I and II and assigned to the selected treatment. The Group I patients were then divided into 3 subgroups and treated with 3 different calcium antagonists (Nifedipine R; Verapamil R and Diltiazem). All group II patients were treated with Enalapril Maleate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546104 TI - [A case of adenovirus-induced hepatitis]. AB - A case of acute adenovirus-induced hepatitis is reported. The diagnosis was based on the increase in adenovirus antibodies and the absence of markers for HBV, HAV and other minor hepatitis viruses on a double serum sample. NANB hepatitis was excluded on the basis of the patient's history, epidemiological data and clinical course. PMID- 2546105 TI - Synaptic transmission at the Schaffer-CA1 synapse is blocked by 6,7-dinitro quinoxaline-2,3-dione. An in vivo brain dialysis study in the rat. AB - 6,7-Dinitro-quinoxaline-2,3-dione (DNQX, FG 9041), a new non-N-methyl-D-aspartate (NMDA) glutamate receptor antagonist, has been reported to block non-NMDA receptor-mediated excitatory amino acidic responses in cultured neurons. We have perfused this compound in vivo through a dialysis fiber placed in the CA1 regions of anesthetized rats to test its effects on CA1 field-evoked potentials. Perfusions of 25-100 microM DNQX completely abolished field excitatory postsynaptic potentials (EPSP) and orthodromic population spikes (PS). This effect was dose-dependent and was reversed after washing with fresh Krebs-Ringer bicarbonate. Antidromic population spikes and fiber volley potentials were unaffected by perfusions of DNQX up to 100 microM. On the contrary, perfusion of 50 microM D-2-amino-5-phosphonovalerate, a specific NMDA receptor antagonist, left unchanged both field EPSP and orthodromic PS. Results demonstrate that low frequency transmission at the Schaffer collaterals-CA1 synapse is mediated by non NMDA glutamate receptors. PMID- 2546106 TI - Experimental ischemia induces a persistent depolarization blocked by decreased calcium and NMDA antagonists. AB - Early physiological events induced by hypoxia plus low D-glucose were investigated by intracellular recording in the rat hippocampal slice. A rapid intracellular depolarization corresponded to the extracellularly recorded anoxic depolarization. This intracellular depolarization consisted of two pharmacologically distinct components, an initial depolarization and a persistent depolarization. The persistent phase of depolarization was selectively blocked by lowering calcium and raising magnesium and by N-methyl-D-aspartate (NMDA) antagonists. This persistent depolarization can account for the long-term synaptic failure seen following experimental ischemia in vitro. PMID- 2546107 TI - Glycine action on N-methyl-D-aspartate receptors in rat hippocampal neurons. AB - The modulatory effect of glycine and other amino acids on the activity of N methyl-D-aspartate (NMDA) receptors has been investigated using a concentration clamp method in the acutely isolated pyramidal neurons from rat hippocampus. The dose-response curves for L-aspartate, measured on the background of several glycine concentrations, were close to Langmuir isotherms with Kd values practically independent of the concentration of glycine. The facilitatory action of glycine appeared at concentrations below 0.1 microM and saturated between 10 and 100 microM. This effect demonstrated marked densensitization, at least at concentrations higher than 5 microM. Facilitation of the responses was shared by amino acids with the potencies diminishing as follows: glycine, D-serine, D alanine greater than L-proline, D-proline, hydroxy-L-proline, taurine greater than L-alanine, L-serine. This sequence did not correlate with the ability of these amino acids to activate strychnine-sensitive glycine receptors. PMID- 2546108 TI - Correlation between regional and kinetic heterogeneities of beta carboline/benzodiazepine receptor binding in rat brain. AB - Kinetic heterogeneity of [3H]methyl beta-carboline-3-carboxylate (CCM) binding was used to distinguish two populations of benzodiazepine binding sites in synaptic membranes of rat cerebral cortex. Curvilinear dissociation plots of CCM binding revealed major high affinity and minor lower affinity populations with slow and rapid dissociation rates, respectively. beta-Carbolines showed some selectivity to displace the higher affinity [3H]CCM binding. The selectivity decreased in the following order: CCM (inverse agonist), FG 7142 (partial inverse agonist), ZK 93426 (antagonist) and ZK 93423 (agonist). Regional selectivity of displacing potencies of these beta-carbolines on [3H]flunitrazepam binding in cerebellar versus hippocampal membranes decreased similarly. PMID- 2546109 TI - Colocalization of NGF receptor immunoreactivity and [3H]GABA uptake activity in developing rat septum/diagonal band neurons in vitro. AB - Neurons in the septum (SEP) and vertical limb of the diagonal band (vDB) of 6-day old rats were dissociated and cultured for 3 days. Cells with high affinity uptake mechanism for gamma-aminobutyric acid (GABA) were identified autoradiographically using 16 nM [3H]GABA. Cells that expressed nerve growth factor (NGF) receptor immunoreactivity were identified immunocytochemically using MC192, a monoclonal antibody against NGF receptor. Double labeling experiments combining [3H]GABA uptake with immunostaining for NGF receptor showed that 46% of the NGF receptor-immunoreactive cells accumulated [3H]GABA. The result was discussed in relation to a possible involvement of the developing septal GABAergic neurons in NGF-induced neuronal survival. PMID- 2546110 TI - Proton-induced Na+ current develops prior to voltage-dependent Na+ and Ca2+ currents in neuronal precursor cells from chick dorsal root ganglion. AB - Proton-induced Na+ currents (INa(H] were measured in precursor cells from chick dorsal root ganglia with whole-cell patch-clamp recording. Precursor cells were isolated using the method of Rohrer et al. (EMBO. J., 4 (1985) 1709-1714) and fast pH changes were applied with a technique developed by Davies et al. (J. Physiol. (Lond.), 400 (1988) 159-187). Proton-induced transient Na+ currents showing the same properties as in more mature neurons could be elicited already early in differentiation, before high-voltage activated Ca2+ currents and voltage dependent Na+ currents develop. PMID- 2546111 TI - Urodynamic changes following hormonal replacement therapy in women with premature ovarian failure. AB - The mechanism by which estrogen supplementation improves the symptom of stress incontinence in postmenopausal women is unclear. Six women with proved premature ovarian failure were studied urodynamically before and after administration of oral and vaginal estrogen to study estradiol's effects on lower urinary tract function. Regardless of the mode of administration, estrogen supplementation did not produce any significant change in urethral pressure, functional length, or cystometric parameters. However, a significant increase in pressure transmission ratio (P less than .05) to the proximal and mid-urethra was noted after the administration of vaginal estrogen cream. We conclude that estrogen alone, in the absence of aging and other known precipitating factors for stress incontinence, is of minimal significance in maintaining normal urinary tract physiology. PMID- 2546112 TI - Hormonal therapy of endometriosis. AB - Hormonal therapy has long been the mainstay of the medical management of endometriosis. However, there is considerable confusion regarding the appropriate application of hormones in the treatment of endometriosis. This article critically reviews the current status of the steroidal therapy of endometriosis. PMID- 2546113 TI - Enzyme-histochemical visualization of lymphatic capillaries in the mouse tongue: light and electron microscopic study. AB - The distinction between lymphatic capillaries and blood capillaries in the mouse tongue was studied enzyme-histochemically by light and electron microscopy. The lymphatic walls are characterized by a strong 5'-nucleotidase (5'-Nase) activity, whereas those of the blood capillaries reveal a significantly lower or no activity. The alkaline phosphatase (ALPase) activity, on the other hand, is markedly higher in the blood capillaries than in the lymphatic capillaries. The specific reaction of 5'-Nase activity in the lymphatic capillaries is obtained by simultaneous inhibition of ALPase on incubation in a medium (Wachstein and Meisel, 1957) with L-tetramisole for 5'-Nase histochemistry. The distribution and intensity of 5'-Nase activity in the lymphatic capillaries can be adequately visualized by comparison with serial cryostat sections for histochemical detection under light and backscattered imaging scanning electron microscopes. The reaction products of the 5'-Nase activity are localized on the outer surface of the cell membrane of the lymphatic endothelial cells, whereas those in the blood capillaries reveal a weak or no reaction. The present results demonstrates satisfactory isolated visualizations of 5'-Nase activity in the lymphatic capillaries and of ALPase activity in the blood capillaries. PMID- 2546114 TI - Glomus vagale tumor: the significance of early vocal cord paralysis. AB - Glomus body tumors most frequently originate in the middle ear (tympanicum) or on the jugular bulb (jugulare). Tumors that arise from the vagal body account for less than 2.5% of these unique paraganglionic neoplasms. Otologic manifestations of tympanicum and jugulare tumors usually precede or accompany neurologic findings. In reviewing five cases of glomus vagale tumors, the initial symptom of voice change, caused by vocal cord paralysis, preceded the presenting symptoms of hearing loss and tinnitus by an average of 2.5 years. The concept of early cranial nerve involvement by glomus vagale tumors is supported in a literature review. The evaluation of "idiopathic" vocal cord paralysis must include radiographic inspection of the skull base, whereas an accurate, temporal account of presenting symptoms may help distinguish vagal body tumors from other vascular neoplasms of this region. PMID- 2546115 TI - Association of DNA aneuploidy with human papillomavirus-induced malignant transformation of sinonasal transitional papillomas. AB - The nuclear DNA content of 19 transitional papillomas of the sinonasal region and 9 maxillary squamous cell carcinomas was studied by flow cytometry; the presence of human papillomavirus (HPV) DNA types 11 and 16 was determined by the in situ hybridization technique from paraffin-embedded tissue. Thirteen (68%) of the papillomas and none of the carcinomas contained HPV genome. Six (32%) of the papillomas and 4 (44%) of the carcinomas had an aneuploid DNA content. The relative DNA content (DNA index) of the aneuploid maxillary carcinomas was larger than that of aneuploid papillomas (p = 0.004). Three of the papillomas underwent malignant transformation, all three of which contained HPV type 16 DNA; two were also aneuploid. Data indicate that papillomas containing HPV type 16 DNA have a tendency (p = 0.06) to undergo malignant transformation, and that this tendency is greater if DNA aneuploidy or HPV type 11 DNA is also present (p = 0.02). PMID- 2546116 TI - A paraneoplastic syndrome associated with glomus tumors of the skull base? Early observations. AB - Independent secretion of vasoactive substances by glomus tumors of the skull base is widely recognized. Surgical removal of these tumors often results in an unexplained prolonged postoperative ileus, even in cases in which the vagus nerve is preserved. There is evidence that these tumors may secrete neuropeptides, such as cholecystokinin, in addition to catecholamines. A retrospective analysis of cases of glomus tumors of the skull base operated on at The Otology Group was carried out to correlate preoperative neuropeptide levels, vagus nerve status at surgery, and duration of postoperative ileus. High circulating levels of cholecystokinin associated with these tumors may be responsible for the unexplained phenomenon of prolonged postoperative ileus. The relevance of neuropeptides to the postoperative management of these patients is discussed. Preventive measures that may avert the potentially lethal complications of aspiration and negative nitrogen balance are described. PMID- 2546117 TI - Pleomorphic adenoma of the nasal septum. PMID- 2546118 TI - Effect of cyclic adenosine monophosphate elevations on functional responses of polymorphonuclear leukocytes from patients with cystic fibrosis. AB - Recently, it has been suggested that the primary metabolic defect of cystic fibrosis (CF) is in the metabolic steps between an elevation of cyclic adenosine monophosphate (cAMP) and the opening of the Cl- ion channel in airway epithelial cells. Results from other studies have indicated that the primary defect may also be present in polymorphonuclear leukocytes (PMNL). In PMNL, cAMP is a potent inhibitor of cell function. We therefore used three compounds (epinephrine, theophylline, and forskolin) known to elevate cAMP in PMNL by three different mechanisms, to study the effect of an elevation of cAMP on various cell functions, such as superoxide production, membrane depolarization, and degranulation of CF patient PMNL and PMNL from healthy individuals. The baseline rates of superoxide production, degranulation, and membrane depolarization were identical for CF and control PMNL. After exposure to epinephrine as well as theophylline plus forskolin, the cell function of CF PMNL was inhibited to the same extent as that of control PMNL. Our results argue against an abnormal response of PMNL from patients with CF to an elevation of cAMP. PMID- 2546119 TI - Follow-up imaging of benign pediatric liver tumors. AB - Though surgery has been recommended in the past for the management of benign hepatic tumors, the current emphasis is on medical management. This report of 4 cases with long-term (8-28 months) follow-up emphasizes the natural history of these tumors which serves to indicate the advantages and disadvantages of CT and ultrasound. The lesions typically involute with some calcification. Calcification can technically be a problem in follow-up with sonography. PMID- 2546120 TI - Lack of association of Haemophilus influenzae b and cytomegalovirus infections. PMID- 2546121 TI - Enteric adenovirus infection and childhood diarrhea: an epidemiologic study in three clinical settings. AB - During a 2-year prospective study of gastroenteritis in children less than 2 years of age, the role of enteric adenovirus as a cause of infantile diarrhea was examined in three clinical settings in a case-control fashion. Using a monoclonal antibody-based enzyme-linked immunosorbent assay with specificity for adenovirus serotypes 40 and 41, enteric adenovirus was identified in 10 of 246 episodes of diarrhea in outpatients (4.1%), 13 of 211 children admitted to the hospital with diarrhea (6.2%), and 5 of 81 children in whom nosocomial diarrhea developed (6.2%), making this agent the third most commonly identified etiologic agent of diarrheal disease. Asymptomatic infections were uncommon (5 of 372 control subjects, or 1.3%) and were seen most frequently in the nosocomial setting. Cases occurred in every calendar month except March and April of each year. A syndrome of watery diarrhea of longer duration compared with other patients with diarrhea (mean 5.4 vs 3.8 days, P = .01), associated with vomiting and dehydration, was present in most cases. Compared with patients with rotavirus, patients were as likely to experience fever and dehydration and more likely to vomit. Household contact with gastroenteritis, often with a child 2 to 5 years of age, was a predisposing factor. It was concluded that enteric adenovirus is an important cause of infantile diarrhea in Baltimore children. Although far less common than rotavirus, this agent was associated with diarrheal illnesses that were at least as severe as those seen with rotavirus. PMID- 2546122 TI - Observations on the effect of an excitatory-inhibitory stimulus on the landing reaction of the fly, Calliphoridae. AB - This research investigates at a behavioral level the sensitivity to movement of the visual system of Calliphora, inferred from the motor response of the landing reaction, to observe the variations of the threshold of velocity of the landing reaction during simultaneous introduction of both excitatory (landing) and inhibitory (antilanding) stimuli. Analysis supports the hypothesis that the interaction of opposing stimuli decreases the sensitivity to movement. Moreover, no effect was observed when the number of stimuli per unit time increased. PMID- 2546123 TI - Highly inducible expression from vectors containing multiple GRE's in CHO cells overexpressing the glucocorticoid receptor. AB - A conditional glucocorticoid-responsive expression vector system is described for highly inducible expression of heterologous genes in mammalian cells. This host vector system requires high level expression of the glucocorticoid receptor (GR) protein in the host cell and multiple copies of the receptor binding site within the expression vector. Transfection and selection of Chinese hamster ovary cells with expression vectors encoding the rat GR yielded cell lines which express functional receptor at high levels. Insertion of multiple copies of the MMTV enhancer (glucocorticoid responsive element, GRE) into an Adenovirus major late promoter (AdMLP) based expression vector yielded greater than 1000-fold inducible expression by dexamethasone (dex) in transient DNA transfection assays. The induced expression level was 7-fold greater than that obtained with an AdMLP based vector containing an SV40 enhancer, but lacking GRE's. Vectors containing the SV40 enhancer in combination with multiple GRE's exhibited elevated basal expression in the absence of dex, but retained inducibility in both transient assays and after integration and amplification in the CHO genome. This expression system should be of general utility for studying gene regulation and for expressing heterologous genes in a regulatable fashion. PMID- 2546124 TI - DNA nucleotide sequence analysis of the immediate-early gene of pseudorabies virus. AB - The complete DNA sequence coding for the immediate-early protein (IE180) of pseudorabies virus was determined. The coding region of IE180 is 4380 nucleotides for 1460 amino acid residues. G+C content of the non-coding portion of the IE gene is 70.3% while the G+C content of the coding portion is considerably higher at 80.1%. Correspondingly, codons consisting mainly of Gs and Cs are favoured. Clusters of amino acid homologies are observed among IE180 of pseudorabies virus, ICP4 of herpes simplex virus type-1 and IE140 of varicella-zoster virus, and are organized similarly in all three polypeptides. Functions exhibited by IE180 are assigned, tentatively, to structural domains of the molecule by analogy to the HSV-1 ICP4 polypeptide. PMID- 2546126 TI - Bioactivity of growth hormone releasing hormone (1-29) analogues after SC injection in man. AB - The bioactivity of growth hormone releasing hormone 1-29 [GHRH(1-29)NH2] has been compared with that of an agonist analogue [Ac-D-Tyr1,D-Ala2]-GHRH(1-29)NH2, in normal male volunteers. Using a submaximal dose of 3 micrograms/kg administered subcutaneously, peak growth hormone (GH) response and area under the GH curve were similar for the native and agonist analogue. In addition, no significant differences were found in peak GHRH(1-29) immunoreactivity, area under the GHRH(1 29) curves or plasma disappearance rates of the two peptides. The results suggest that, in keeping with the relative activities of other "superactive" analogues tested so far, the greatly enhanced activity of [Ac-D-Tyr1,D-Ala2]-GHRH(1-29)NH2 observed in the rat is not found in humans. It is possible that this species difference is due to differences in the interaction of GHRH peptides with the rat and the human somatotroph GHRH receptor. PMID- 2546125 TI - Two related superfamilies of putative helicases involved in replication, recombination, repair and expression of DNA and RNA genomes. AB - In the course of systematic analysis of protein sequences containing the purine NTP-binding motif, a new superfamily was delineated which included 25 established or putative helicases of Escherichia coli, yeast, insects, mammals, pox- and herpesviruses, a yeast mitochondrial plasmid and three groups of positive strand RNA viruses. These proteins contained 7 distinct highly conserved segments two of which corresponded to the "A" and "B" sites of the NTP-binding motif. Typical of the new superfamily was an abridged consensus for the "A" site, GxGKS/T, instead of the classical G/AxxxxGKS/T. Secondary structure predictions indicated that each of the conserved segments might constitute a separate structural unit centering at a beta-turn. All previously characterized mutations impairing the function of the yeast helicase RAD3 in DNA repair mapped to one of the conserved segments. A degree of similarity was revealed between the consensus pattern of conserved amino acid residues derived for the new superfamily and that of another recently described protein superfamily including a different set of prokaryotic, eukaryotic and viral (putative) helicases. PMID- 2546127 TI - D-Phe7-substituted peptide bradykinin antagonists are not substrates for kininase II. AB - The bradykinin receptor antagonists [D-Phe7]bradykinin, D-Arg[Hyp3,D Phe7]bradykinin and D-Arg[Hyp3,Thi5,8,D-Phe7]bradykinin were tested for their ability to serve as substrates for kininase II (angiotensin converting enzyme) purified from rabbit lung. By HPLC, the peptides were not measurably degraded over 30 minutes. Under identical conditions, bradykinin was completely degraded to bradykinin (1-7). When hippuryl-His-Leu was used as a substrate for kininase II, the D-Phe7-substituted bradykinins acted as weak noncompetitive inhibitors. While the peptides were poor substrates for kininase II, they were short-lived when injected intravenously. D-Arg[Hyp3,D-Phe7]bradykinin was completely degraded to small fragments in less than 2 minutes. In diluted serum in vitro, a single product was observed with elution consistent with loss of arginine, suggestive of metabolism by kininase I. PMID- 2546129 TI - Chronic ACTH treatment: influence on 5-HT2 receptors and behavioral supersensitivity induced by 5,7-dihydroxytryptamine lesions. AB - The capacity of the serotonin (5-HT) precursor 5-HIP to induce the ACTH responsive myoclonic-convulsive disorder infantile spasms in patients with Down's syndrome has been cited as evidence for altered serotonergic neurotransmission in infantile spasms. Since there is no animal model of infantile spasms, the suitability of behavioral supersensitivity (myoclonus) evoked by 5-HTP in rats with 5,7-dihydroxytryptamine (DHT) lesions as a model was tested by determining the effect of chronic treatment with ACTH (40 IU/kg) on 5-HTP-evoked myoclonus. In rats treated with DHT as adults, ACTH administration did not alter the "serotonergic behaviors," such as myoclonus, induced by 30 mg/kg 5 hydroxytryptophan (5-HTP), but induced a small significant increase in Bmax of neocortical 5-HT2 sites of the DHT group, with no change in rats without lesions. In rats treated with DHT as neonates, there was also no significant difference in behaviors evoked by several doses of 5-HTP. These data suggest that ACTH minimally modifies the effects on 5-HT receptors of DHT lesions, but the intracisternal DHT model is not a suitable model for infantile spasms because chronic ACTH was not antimyoclonic. PMID- 2546128 TI - Corticotropin-releasing factor (CRF) receptors in intermediate lobe of the pituitary: biochemical characterization and autoradiographic localization. AB - CRF receptors were characterized using radioligand binding and chemical affinity cross-linking techniques and localized using autoradiographic techniques in porcine, bovine and rat pituitaries. The binding of 125I-[Tyr0]-ovine CRF (125I oCRF) to porcine anterior and neurointermediate lobe membranes was saturable and of high affinity with comparable KD values (200-600 pM) and receptor densities (100-200 fmoles/mg protein). The pharmacological rank order of potencies for various analogs and fragments of CRF in inhibiting 125I-oCRF binding in neurointermediate lobe was characteristic of the well-established CRF receptor in anterior pituitary. Furthermore, the binding of 125I-oCRF to both anterior and neurointermediate lobes of the pituitary was guanine nucleotide-sensitive. Affinity cross-linking studies revealed that the molecular weight of the CRF binding protein in rat intermediate lobe was identical to that in rat anterior lobe (Mr = 75,000). While the CRF binding protein in the anterior lobes of porcine and bovine pituitaries had identical molecular weights to CRF receptors in rat pituitary (Mr = 75,000), the molecular weight of the CRF binding protein in porcine and bovine intermediate lobe was slightly higher (Mr = 78,000). Pituitary autoradiograms from the three species showed specific binding sites for 125I-oCRF in anterior and intermediate lobes, with none being apparent in the posterior pituitary. The identification of CRF receptors in the intermediate lobe with comparable characteristics to those previously identified in the anterior pituitary substantiate further the physiological role of CRF in regulating intermediate lobe hormone secretion. PMID- 2546131 TI - Rat renal mesenchymal tumor as an experimental model for human congenital mesoblastic nephroma: I. Induction. AB - The effect of low-dose (2 Gy) radiation on ethylnitrosourea (ENU)-induced neoplasms was studied in Sprague-Dawley and Holtzman strains of rats. With a 60 mg/kg dose of ENU administered on day 1 in Sprague-Dawley rats, 18.4% of the neoplasms induced were found in the kidney. When the same dose of ENU was given on day 10, the incidence of kidney tumors fell to 2.8%. Prior (2 Gy) radiation on day 9 enhanced kidney tumor induction to 16.1%, a trend also observed in the case of ENU-induced neural tumors. In Holtzman rats, 40 mg/kg ENU induced more kidney tumors (12.5%) when given on day 4 than on day 0, and prior irradiation enhanced the ENU-induced kidney tumors even though the interval between irradiation and carcinogen administration was fairly long--4 days. PMID- 2546130 TI - Inflammatory fibrous histiocytoma presenting leukemoid reaction. AB - A case of inflammatory fibrous histiocytoma presenting leukemoid reaction is reported. A tumor had developed at the right scapular region 10 years ago and recurred twice with fever and leukocytosis, and finally with leukemoid reaction. Besides the histopathological features typical of malignant fibrous histiocytoma, this tumor showed prominent and diffuse infiltration of neutrophilic leukocytes. Hematologic examination of the bone marrow aspirates disclosed myeloid hyperplasia. Bacteriological examination revealed no pathogenic microorganisms in the tumor tissue. Chemotactic activity of the neutrophilic leukocytes was demonstrated in the extract obtained from the tumor, and subcutaneous injection of the extract into rats induced myeloid hyperplasia in the bone marrow and neutrophil infiltration in the liver and spleen of the animals. These results suggest the possibility of the case as a colony-stimulating factor-producing tumor. PMID- 2546132 TI - Rat renal mesenchymal tumor as an experimental model for human congenital mesoblastic nephroma: II. Comparative pathology. AB - The morphological features of kidney neoplasms induced by ethylnitrosourea (ENU) with or without prior irradiation were examined with a view to comparing them with human renal tumors of childhood. The rat renal tumors consisted of poorly differentiated, highly mitotic mesenchymal cells frequently attempting to differentiate along fibroblastic and myofibroblastic lineages. Though the host renal tubules were frequently entrapped in these tumors, immature tubules and islands of epithelial cells occasionally were seen to form an integral part of the neoplasms. Rarely, adenomatous areas surrounded by mesenchymal proliferation were observed. None of the tumors had the blastemal component that is the hallmark of nephroblastoma. The rat mesenchymal tumors resembled the human congenital mesoblastic nephroma. PMID- 2546133 TI - Monoclonal antibody MB2: a potential marker for Ewing's sarcoma and primitive neuroectodermal tumor. AB - The small round-cell tumors of childhood present difficulties in diagnosis when differentiation is not apparent. Immunohistochemistry is helpful; however, the only antigen consistently detected in Ewing's sarcoma is vimentin, which may also be detected in the other types of small-cell neoplasms. The monoclonal antibody (MAb) MB2 is marketed as a B-lymphocyte marker that can be used on paraffin embedded tissue. To determine its specificity, we performed immunohistochemical staining on pediatric tumors with MB2. These included 55 cases of small round cell tumors (lymphomas, Ewing's sarcoma, peripheral primitive neuroectodermal tumors [PNET], neuroblastomas, rhabdomyosarcomas, and nephroblastomas). MB2 positivity was detected in all B-cell lymphomas and in seven of nine cases of Ewing's sarcoma and three PNET. In neuroblastomas only differentiating ganglion cells were positive. In rhabdomyosarcomas only large rhabdomyoblasts were positive. Blastema of nephroblastomas was negative. Thus, in cases of poorly differentiated small round-cell tumors, MB2 was positive in all B-cell lymphomas, most Ewing's sarcomas and all cases of PNET. Lymphomas were distinguished by staining for leukocyte-common antigen and PNET by neuron-specific enolase. Therefore, the addition of MB2 to a discrete panel of antibodies may prove useful in the diagnosis of Ewing's sarcoma and PNET. PMID- 2546134 TI - Case 1. Postnatally acquired cytomegalovirus hepatitis. PMID- 2546135 TI - Case 5. Adenovirus pneumonia in a newborn. PMID- 2546136 TI - 4(del p) syndrome and cytomegalovirus. PMID- 2546137 TI - Primitive neuroectodermal tumor of the brain associated with malignant rhabdoid tumor of the liver: a histologic, immunohistochemical, and electron microscopic study. AB - A 14-day-old white male, born with a large primitive neuroectodermal tumor of the left cerebral hemisphere, was found to have a solitary rhabdoid tumor in the liver incidentally at autopsy. Cells resembling the liver rhabdoid cells were also found by histology, immunohistochemistry, and electron microscopy in the brain tumor. The concurrence of rhabdoid cells in the tumors of the brain and liver suggests a common histogenesis and further supports the previous suggestion that the rhabdoid tumor is of neuroectodermal origin. The rhabdoid tumor in the liver in this case is likely to be a metastatic tumor from the brain rather than a second primary tumor. PMID- 2546138 TI - Lipolytic activities of p-aminophenol and n-decylamine in endogenous lipid droplets from rat adipocytes. AB - Lipolytic agents such as adrenaline (epinephrine), dibutyryl cyclic AMP (DBcAMP) and theophylline possess common structural characteristics, the existence of a positive charge and a hydrophobic area. p-Aminophenol, which has the same structural characteristics, was found to induce lipolysis in the endogenous lipid droplets from rat adipocytes. It was demonstrated that the positive group and the hydrophobic area of p-aminophenol are essential requirements for its lipolytic activity. Among various amine derivatives, only n-decylamine was found to induce lipolysis in the endogenous lipid droplets. Although the lipolytic action of n decylamine is fairly low, it possesses remarkable inhibitory effects on both adrenaline- and DBcAMP-induced lipolytic activities. Furthermore, n-decylamine is found to inhibit competitively the DBcAMP-induced lipolysis. PMID- 2546139 TI - Effect of praziquantel on some aspects of carbohydrate metabolism in mice infected with Schistosoma mansoni. AB - Schistosoma mansoni infection in mice resulted in a marked decrease in blood glucose and liver glycogen accompanied by a significant increase in hepatic glucose-6-phosphatase (G-6-Pase) activity. Moreover, the results indicated that infection produced a significant increase in blood pyruvate and hepatic glucose-6 phosphate dehydrogenase (G-6-PD) activity with a significant decrease in blood lactate. Infected mice were treated with praziquantel which was given at two doses of 500 mg/kg body wt on two consecutive days. Seven and 14 days respectively after drug administration, such treatment caused a marked improvement in the previous aspects of carbohydrate metabolism. This is indicated by the tendency of the blood glucose of infected mice to be restored, the marked increase in their liver glycogen content, the normalization of their blood lactate and pyruvate as well as by the marked decrease of their hepatic G-6-Pase activity and the progressive increase in their hepatic G-6-PD activity. Praziquantel given to normal mice moderately affected the blood glucose and the previously mentioned hepatic enzymes. However, the drug markedly increased the liver glycogen content of normal mice and failed to elicit any change in their blood pyruvate and lactate. Possible explanations of these findings are discussed. PMID- 2546140 TI - Influence of PLA2-PG system on purine release and cAMP content in dissociated primary glial cultures from rat striatum. AB - Purine release and prostaglandin (PG) outflow were simultaneously evaluated from untreated glial primary cultures of rat striatum, at rest and under field electrical stimulation. Purine release was also assayed from sister cultured cells in which a suitable pharmacological treatment with 1 x 10(-6) M dexamethasone or 1 x 10(-4) M indomethacin had produced a complete inhibition of the phospholipase A2-prostaglandin (PLA2-PG) system. Purine release from untreated cells seems to be regulated by specific receptor sites for released adenosine (Ado); A1 receptors exert an inhibitory control on purine release while A2 receptors facilitate it. PG release appears to be related to A1-mediated Ado activity, since culture treatment with 1 x 10(-10) M 8-cyclopentyl-1,3 dipropylxanthine (DPCPX) or 1 x 10(-4) M N-ethylmaleimide (NEM), A1 receptor inhibitory agents able to increase purine release, induced a significant reduction of the evoked PG outflow. Purine amount, released from glial cells with inhibited PLA2-PG system, was remarkably greater than that one assayed from control cultured cells. In so treated cultures, no additive effect, NEM-induced, was detected, while the addition of a mixture of PGs partially reduced the increased purine outflow. An electrically evoked cAMP accumulation, significantly greater than that found in controls, was even detected in cultured cells with inhibited PLA2-PG system. Since 10 micrograms/ml adenosine deaminase (ADA) reduced while DPCPX enhanced the evoked cAMP accumulation, it seems partially due to released Ado and accounts for a prevalent A2-stimulating rather than an A1 inhibitory control on adenylate cyclase activity. Thus, in cultured glial cells, the PLA2-PG system, likely linked to A1 receptor sites, concurs to control purine release and seems to affect less directly cAMP accumulation. PMID- 2546141 TI - Transport characteristics of ceftibuten (7432-S), a new oral cephem, in rat intestinal brush-border membrane vesicles: proton-coupled and stereoselective transport of ceftibuten. AB - The transport characteristics of ceftibuten in rat intestinal brush-border membrane vesicles were investigated by a rapid filtration technique. Ceftibuten uptake was markedly stimulated by an inwardly directed H+ gradient (pH 7.5 inside, pH 5.5 outside) in comparison with that in the absence of a H+ gradient. The uptake at 30 sec was four times greater than that observed at equilibrium (overshoot phenomenon), while the H+ gradient-stimulated uptake of ceftibuten was markedly reduced in the presence of FCCP, a protonophore. These results suggested H+-coupled uphill transport of ceftibuten. In contrast, an inwardly directed Na+ gradient had no effect on ceftibuten uptake. The valinomycin-induced K+ diffusion potential (inside positive) significantly stimulated the ceftibuten uptake, suggesting net transfer of the negative charge. In contrast to the cis-isomer ceftibuten, the trans isomer of ceftibuten is not readily absorbed from the intestine, and its uptake was found not to be affected by a H+ gradient. Since the lipophilicity of the trans isomer is similar to that of ceftibuten, the uptake process appears to be stereoselective. The initial uptake of ceftibuten and its analogue cefaclor was concentration dependent under a H+ gradient. The apparent Km value was 0.2 mM for ceftibuten and 3.0 mM for cefaclor. PMID- 2546142 TI - Kinetics of the aspartyl transpeptidation of daptomycin, a novel lipopeptide antibiotic. AB - Two degradation products of the lipopeptide antibiotic, daptomycin, were identified and the reaction pathway and kinetics were delineated in aqueous solution at 60 degrees C, pH range 3 to 8 and ionic strength 0.01. The degradation products were 1) a succinimido intermediate (anhydro-daptomycin) formed by attack of side-chain carbonyl on the peptide-bond nitrogen in the asp gly sequence and 2) a beta-asp daptomycin isomer formed by rehydration of the anhydrodaptomycin succinimide. This aspartyl transpeptidation pathway was found to be reversible. Formation of the anhydrodaptomycin from either daptomycin or beta-asp daptomycin was pH dependent but the pH-rate profiles for anhydrodaptomycin formation were not mechanistically interpretable. The pH-rate profiles for the formation of daptomycin or beta-asp daptomycin from the anhydrodaptomycin were linear with slopes = 1, which is consistent with nucleophilic hydroxide ion attack of the succinimido intermediate at either the alpha-carbonyl, giving rise to the beta-asp daptomycin, or the beta-carbonyl, giving rise to daptomycin. PMID- 2546143 TI - Efficiency of arsenic clearance from human blood in vitro and from dogs in vivo by extracorporeal complexing haemodialysis. AB - In vitro dialysis using human blood and in vivo extracorporeal haemodialysis trials with dogs, in the presence and absence of various chelating agents, showed that haemodialysis alone facilitated efficient removal of the metal, even in the absence of any of the chelating agents. In vitro haemodialysis for 5 hr without chelating agents caused the removal of 85% of the arsenic (As) added to the blood 1 hr before the beginning of the treatment. Ten-33% of the single dose given to the dogs, were removed into the dialysate during the first treatment and 8-15% during the second. The amounts of As excreted in the urine of dogs given 0.25 mg As/kg b.wt./day for 6 days were 6.8-7.4 mg As/24 hr, while the same amounts of the metal were removed into the dialysate during haemodialysis for 5 hr. This demonstrates the advantage of employing haemodialysis over the conventional treatment for the enhanced removal of As from the body. PMID- 2546144 TI - Tissue distribution of glyceryl trinitrate and the effect on cGMP levels in rat. AB - In order to study distribution in tissue, rats were treated subcutaneously with glyceryl trinitrate, GTN, (50 mg/kg). The concentrations of GTN were measured in plasma, brain, heart, adipose tissue and aortic tissue at different sampling times by a gas chromatographic method with electron-capture detection. The peak GTN-concentration was reached after 2 hours in all tissues examined. The highest concentration of GTN was found in adipose tissue, where the level was approximately forty times higher than in plasma. The concentration of GTN in brain, heart and aortic tissue was about 2-3 times as high as in plasma. The cGMP level was measured in heart and brain. An increase of the cGMP level was found in brain 2 hours after GTN administration. No cGMP increase was found in heart tissue. The results indicate a substantial distribution of GTN to tissue, and an increase of cGMP in brain. The distribution of the substance in the tissues as shown, might have both pharmacokinetic and pharmacodynamic implications. PMID- 2546145 TI - Histidine-15: an important role in the cytotoxic activity of human tumor necrosis factor. AB - The amino acids that are required for the cytotoxic activity of recombinant human tumor necrosis factor-alpha (TNF) were investigated by chemical modification and oligonucleotide-directed site-specific mutagenesis. TNF contains three histidine residues, located at positions 15, 73 and 78. The histidine-specific reagent diethylpyrocarbonate (DEP) was used to chemically modify TNF. The chemical inactivation of the in vitro cytotoxic activity of this lymphokine (using murine L929 target cells) was found to be time- and dose-dependent. Inactivated TNF failed to compete with fully bioactive [125I]TNF for human MCF-7 target cell receptors. Mutant polypeptides of TNF were genetically engineered by oligonucoleotide-directed site-specific mutagenesis. The cytotoxicity of a double histidine mutant, in which histidine-73 and histidine-78 were replaced with glutamine, was not altered and was chemically inactivated by DEP. Substituting glutamine for histidine-15 resulted in 10-15% of the wild-type bioactivity. Replacing histidine-15 with either asparagine, lysine or glycine resulted in a biologically inactive molecule. The data show that the histidine residue at position 15 is an amino acid that is required for the cytotoxic activity of TNF. PMID- 2546146 TI - Secondary structural analysis of retrovirus integrase: characterization by circular dichroism and empirical prediction methods. AB - The retrovirus integrase (IN) protein is essential for integration of viral DNA into host DNA. The secondary structure of the purified IN protein from avian myeloblastosis virus was investigated by both circular dichroism (CD) spectroscopy and five empirical prediction methods. The secondary structures determined from the resolving of CD spectra through a least-squares curve fitting procedure were compared with those predicted from four statistical methods, e.g., the Chou-Fasman, Garnier-Osguthorpe-Robson, Nishikawa-Ooi, and a JOINT scheme which combined all three of these methods, plus a pure a priori one, the Ptitsyn Finkelstein method. Among all of the methods used, the Nishikawa-Ooi prediction gave the closest match in the composition of secondary structure to the CD result, although the other methods each correctly predicted one or more secondary structural group. Most of the alpha-helix and beta-sheet states predicted by the Ptitsyn-Finkelstein method were in accord with the Nishikawa-Ooi method. Secondary structural predictions by the Nishikawa-Ooi method were extended further to include IN proteins from four phylogenetic distinct retroviruses. The structural relationships between the four most conserved amino acid blocks of these IN proteins were compared using sequence homology and secondary structure predictions. PMID- 2546147 TI - Monoclonal antibodies to the insulin receptor mimic metabolic effects of insulin but do not stimulate receptor autophosphorylation in transfected NIH 3T3 fibroblasts. AB - The metabolic actions of insulin and anti-insulin receptor monoclonal antibodies were compared with their effects on insulin receptor phosphorylation in mouse NIH 3T3 fibroblasts transfected with human insulin receptor cDNA. In serum-starved NIH 3T3 HIR3.5 cells, uptake of 2-deoxy-[3H]glucose was stimulated up to 2-fold after 30 min with insulin, with a half-maximal effect at 0.1 nM insulin. Incorporation of [3H]thymidine was stimulated approximately 12-fold after a 16-hr preincubation with insulin, with a half-maximal effect at 2 nM insulin. Phosphorylation of insulin receptor beta-subunit in cells prelabeled with [32P]phosphate was increased 10- to 20-fold within 5 min of adding insulin, with a half-maximal effect at approximately 3 nM insulin. Monoclonal antibodies reacting with four different epitopes on the insulin receptor mimicked the effect of insulin on 2-deoxyglucose uptake. These antibodies also stimulated thymidine incorporation, although the maximum stimulation was only approximately 30% that of insulin. Two antibodies (25-49 and 83-14) showed a similar concentration dependence to insulin in their metabolic effects and in the inhibition of 125I labeled insulin binding to cells. The other two antibodies (83-7 and 18-44) were somewhat less potent and did not inhibit insulin binding. None of the antibodies significantly increased insulin receptor phosphorylation at concentrations up to 100 nM, which at least in the case of 25-49 and 83-14 was sufficient for full receptor occupancy. It is concluded that the insulin-like metabolic effects of antibodies involve a mechanism of receptor activation that is independent of autophosphorylation and hence that receptor autophosphorylation is not an essential step in triggering at least some events in the insulin signaling pathway. PMID- 2546148 TI - The (A)BC excinuclease of Escherichia coli has only the UvrB and UvrC subunits in the incision complex. AB - The uvrA, uvrB, and uvrC genes control excision repair in Escherichia coli. Cells with mutations in any of these three genes cannot repair DNA by nucleotide excision. When the purified gene products--the UvrA, UvrB, and UvrC proteins--are mixed together, an excision nuclease is formed that incises on both sides of the damaged nucleotide in an ATP-dependent reaction; it has been presumed that the excision nuclease was an ABC complex containing all three Uvr proteins. To determine the stoichiometry of the subunits in the enzyme, we conducted hydrodynamic studies with mixtures of the subunits with or without DNA substrate. We found that without DNA the UvrA subunit is a dimer and that when UvrB protein is also present, a (UvrA)2(UvrB)1 complex forms. Without DNA no detectable interaction of either the UvrA or UvrB subunits or the (UvrA)2(UvrB)1 complex with the UvrC subunit occurs. Unexpectedly, with UV-irradiated DNA, the UvrA/UvrB ratio in isolated DNA-protein complexes is variable, and the ratio becomes infinitesimally low as the UvrA concentration in the reaction mixture decreases. Under conditions of saturating UvrB protein approximately one UvrB molecule binds to DNA per damaged site in a reaction that requires catalytic amounts of UvrA subunit. Addition of UvrC protein to purified UvrB-DNA complexes results in rapid incision of the DNA, presumably catalyzed by an excision nuclease containing only UvrB and UvrC subunits. PMID- 2546149 TI - Human placenta protein-tyrosine-phosphatase: amino acid sequence and relationship to a family of receptor-like proteins. AB - The amino acid sequence of the cytosolic human placenta protein-tyrosine phosphatase 1B (PTPase 1B; protein-tyrosine-phosphate phosphohydrolase, EC 3.1.3.48) has been determined. It consists of a single chain of 321 residues with an N-acetylated N-terminal methionine and an unusually proline-rich C-terminal region. The enzyme is structurally related to the two cytoplasmic domains of both the leukocyte common antigen CD45 and LAR, a CD45-like molecule with an external segment that resembles a neural cell adhesion molecule. A low molecular weight protein encoded by a cDNA clone from T cells also shows extensive sequence similarities. The present study defines homologous domains common to this diverse family of PTPases that includes both soluble and receptor-like transmembrane forms. The cysteinyl residues 121 and 215 of PTPase 1B are conserved among all members of the family and are candidates for involvement in catalysis since PTPase 1B is inactivated by thiol modifying reagents. Two segments rich in positively charged residues (residues 33-47 and 227-238) may provide sites of interaction with inhibitory anionic polymers such as heparin or poly(Glu/Tyr). PMID- 2546150 TI - cDNA isolated from a human T-cell library encodes a member of the protein tyrosine-phosphatase family. AB - A human peripheral T-cell cDNA library was screened with two labeled synthetic oligonucleotides encoding regions of a human placenta protein-tyrosine phosphatase (protein-tyrosine-phosphate phosphohydrolase, EC 3.1.3.48). One positive clone was isolated and the nucleotide sequence was determined. It contained 1305 base pairs of open reading frame followed by a TAA stop codon and 978 base pairs of 3' untranslated end, although a poly(A)+ tail was not found. An initiator methionine residue was predicted at position 61, which would result in a protein of 415 amino acid residues (Mr, 48,400). This was supported by the synthesis of a Mr 48,000 protein in an in vitro reticulocyte lysate translation system using RNA transcribed from the cloned cDNA and T7 RNA polymerase. The deduced amino acid sequence was compared to other known proteins revealing 65% identity to the low Mr PTPase 1B isolated from placenta. In view of the high degree of similarity, the T-cell cDNA likely encodes a newly discovered protein tyrosine-phosphatase, thus expanding this family of genes. PMID- 2546151 TI - The v-sea oncogene of avian erythroblastosis retrovirus S13: another member of the protein-tyrosine kinase gene family. AB - The cloning and sequencing of the oncogene of the avian erythroblastosis virus S13 is described. The oncogene, termed v-sea, was found to be another member of the protein-tyrosine kinase gene family. The oncogene was fused in frame with the retrovirus S13 envelope gene, thus generating a fusion protein with a structure resembling that of a growth factor receptor. Sequence comparisons revealed that the v-sea gene was most closely related to the insulin receptor family of protein tyrosine kinases, the greatest similarity being with the human MET oncogene. PMID- 2546152 TI - A third human retinoic acid receptor, hRAR-gamma. AB - Retinoic acid receptors (RARs) are retinoic acid (RA)-inducible enhancer factors belonging to the superfamily of steroid/thyroid nuclear receptors. We have previously characterized two human RAR (hRAR-alpha and hRAR-beta) cDNAs and have recently cloned their murine cognates (mRAR-alpha and mRAR-beta) together with a third RAR (mRAR-gamma) whose RNA was detected predominantly in skin, a well-known target for RA. mRAR-gamma cDNA was used here to clone its human counterpart (hRAR gamma) from a T47D breast cancer cell cDNA library. Using a transient transfection assay in HeLa cells and a reporter gene harboring a synthetic RA responsive element, we demonstrate that hRAR-gamma cDNA indeed encodes a RA inducible transcriptional trans-activator. Interestingly, comparisons of the amino acid sequences of all six human and mouse RARs indicate that the interspecies conservation of a given member of the RAR subfamily (either alpha, beta, or gamma) is much higher than the conservation of all three receptors within a given species. These observations indicate that RAR-alpha, -beta, and gamma may perform specific functions. We show also that hRAR-gamma RNA is the predominant RAR RNA species in human skin, which suggests that hRAR-gamma mediates some of the retinoid effects in this tissue. PMID- 2546153 TI - Molecular cloning of rat homologues of the Drosophila melanogaster dunce cAMP phosphodiesterase: evidence for a family of genes. AB - To study the structure and function of cyclic nucleotide phosphodiesterases (PDEs) involved in mammalian gametogenesis, a rat testis cDNA library was screened at low stringency with a cDNA clone coding for the Drosophila melanogaster dunce-encoded PDE as a probe. This screening resulted in the isolation of two groups of cDNA clones, differing in their nucleotide sequences (ratPDE1 and ratPDE2). In the rat testis, RNA transcripts corresponding to both groups of clones were expressed predominantly in germ cells. Additional screenings of a Sertoli cell cDNA library with a ratPDE2 clone as a probe led to the isolation of two more groups of clones (rat-PDE3 and ratPDE4). Unlike ratPDE1 and ratPDE2, these clones hybridized to transcripts present predominantly in the Sertoli cell. In the middle of the coding region, all four groups of clones were homologous to each other. The deduced amino acid sequences of part of this region were also homologous to the D. melanogaster dunce PDE and to PDEs from bovine and yeast. These data indicate that a family of genes homologous to the D. melanogaster dunce-encoded PDE is present in the rat and that these genes are differentially expressed in somatic and germ cells of the seminiferous tubule. These findings provide a molecular basis for the observed heterogeneity of cAMP PDEs. PMID- 2546154 TI - Characterization of the termini and transposition products of Tn4399, a conjugal mobilizing transposon of Bacteroides fragilis. AB - We have isolated a 9.6-kilobase conjugal transposon, Tn4399 from Bacteroides fragilis, that is capable of mobilizing nonconjugal plasmids in cis. Here we characterize the ends of the transposon, its target-site requirements, and the products of transposition into the B. fragilis chromosome and two sets of B. fragilis-Escherichia coli shuttle vectors. With the exception of an additional cytosine residue in the left end, there are perfect 13-base-pair (bp) inverted repeats at the ends of Tn4399. Insertion of Tn4399 resulted in a 3-bp target-site repeat in 8 out of 12 independent transpositions and showed a high insertion-site specificity. A remarkable feature of Tn4399 insertions is the presence of an additional 5 bp located between the right inverted repeat and the target-site repeat. Four sequence variations of the 5 bp were found, with absolute conservation at positions 1, 2, and 5. Only two of the variations were present in junction fragments of all three copies of Tn4399 contained in the chromosome of the original donor strain, B. fragilis TM4.2321. Tn4399 appears to represent a new type of conjugal transposon. In contrast to Tn916 and Tn1545, described in streptococci, Tn4399 creates a target-site repeat and contains an additional 5 bp at the right end only, between the transposon and the target sequence. In addition, Tn4399 can mobilize plasmids in cis. PMID- 2546155 TI - Formation of hybrid cell-cell channels. AB - The oocyte cell-cell channel assay was used to demonstrate that connexin-43 is a cell-cell channel-forming protein as previously shown for connexin-32. Expression of connexin-32 in one and connexin-43 in the other oocyte of a pair results in the formation of junctional conductances at rates similar to those observed when only one or the other connexin is expressed in both oocytes of a pair. This suggests that hybrid cell-cell channels form in the oocyte system. Hybrid channels also form when a connexin-43 mRNA-injected oocyte is paired with a noninjected oocyte expressing endogenous connexin. The latter hybrids have properties apparently contributed by both types of hemichannels. Pure connexin-43 channels are not voltage gated, whereas pure oocyte channels are voltage dependent; hybrids of these channels rectify. PMID- 2546157 TI - Diversity in T-cell receptor gamma gene usage in intestinal epithelium. AB - The intraepithelial cells of the murine small intestine include a significant number of CD3+ T cells that use T-cell receptor gamma genes rather than T-cell receptor beta genes. As with other sites of T-cell receptor gamma expression, combinatorial diversity is limited, but there is junctional diversity, and this, together with the specific variable region gamma gene segments used, distinguishes gamma gene expression in the gut epithelium from that in cells derived from the dermal epithelium. The restriction of productive gamma gene expression largely to one V-J-C (V, variable; J, joining; C, constant) gene combination may result from nonproductive joining of other V-J combinations and from productively rearranged genes rendered nonfunctional by incorrect splicing. PMID- 2546158 TI - IgE secretion by Epstein-Barr virus-infected purified human B lymphocytes is stimulated by interleukin 4 and suppressed by interferon gamma. AB - The cytokine interleukin 4 (IL-4) has been shown to induce lipopolysaccharide activated murine B cells to differentiate into IgE-secreting cells and to stimulate IgE secretion by cultured human peripheral blood lymphoid cells. It is unclear, however, whether this effect of IL-4 on human peripheral blood lymphoid cells is a direct effect on the B cell because IL-4 can stimulate T cells and monocytes as well as B cells and does not induce purified human B cells to secrete immunoglobulin. To investigate this issue we studied the ability of IL-4 to induce IgE secretion by purified human B cells (93-96% CD20+, less than 1% CD3+) that were cultured with Epstein-Barr virus (EBV). Although B cells cultured with IL-4 alone did not secrete Ig and B cells cultured with EBV alone secreted IgM, IgG, and IgA but less than 150 pg of IgE per ml, the combination of EBV and IL-4 induced an IgE response that ranged from 11.4 to 40.3 ng/ml of culture supernatant after 26 days of culture. While IL-4 also enhanced IgM, IgG, and IgA secretion, as well as proliferation by EBV-infected B cells, these effects were less pronounced, occurred earlier during culture, and required a lower concentration of IL-4 than did the stimulation of IgE secretion. Furthermore, interferon gamma at 10 units per ml was found to inhibit IL-4/EBV-induced IgE secretion without inhibiting the other stimulatory effects of IL-4. We conclude that (i) IL-4 and interferon gamma can act directly on polyclonally activated human B cells to respectively stimulate and suppress IgE secretion and (ii) IL-4, in addition to its specific effect on IgE secretion, has a general stimulatory effect on the growth and differentiation of EBV-infected human B cells. PMID- 2546156 TI - Dysfunction of chromosomal loop attachment sites: illegitimate recombination linked to matrix association regions and topoisomerase II. AB - A family of A + T-rich sequences termed MARs ("matrix association regions") mediate chromosomal loop attachment. Here we demonstrate that several MARs both specifically bind and contain multiple sites of cleavage by topoisomerase II, a major protein of the mitotic chromosomal scaffold. Interestingly, "hotspots" of enzyme cutting occur within the MAR of the mouse immunoglobulin kappa-chain gene at the breakpoint of a previously described chromosomal translocation. Since topoisomerase II can mediate illegitimate recombination in prokaryotes, we explored further the possibility that MARs might be targets for this process in eukaryotes. We found that a MAR had been deleted from one of the two rabbit immunoglobulin kappa-chain genes and that MARs reside next to a long interspersed repetitive element within the recombination junction of a human ring chromosome 21. These results, taken together with other accounts of nonhomologous recombination, lead to the proposal that a dysfunction of MARs is illegitimate recombination. PMID- 2546159 TI - Retinoid receptor antisense DNAs inhibit alkaline phosphatase induction and clonogenicity in malignant keratinocytes. AB - Antisense oligodeoxynucleotides [oligo(dN)s] corresponding to human cellular retinol-binding protein I (cRBP) and human nuclear retinoic acid receptor alpha (hnRAR) were synthesized. Exposure of human malignant keratinocytes to these oligo(dN)s significantly attenuated the level of cytoplasmic cRBP and hnRAR in a concentration- and time-dependent manner. Further, the induction of alkaline phosphatase by retinol in these cells was blocked by treatment with 30 microM antisense oligo(dN) to cRBP or hnRAR but not by 30 microM of sense oligo(dN) to cRBP. Antisense oligo(dN) treatments concomitantly induced cell rounding, loss of cell-cell attachment, and cell adhesion to the substratum. By contrast, treatment of cells with an anticytokinetic agent, cytochalasin B, or with a cytostatic concentration of sodium azide failed to reduce cytoplasmic cRBP or hnRAR from nuclear extracts, even though antisense oligo(dN)-like changes in cell morphology were observed. Treatment of the cells for greater than 2.75 hr with 20-40 microM of either antisense oligo(dN) also led to the loss of clonogenic potential. These results show that both cytoplasmic and nuclear receptors for retinoids are important in the transduction of a retinoid signal response critical to cellular growth and differentiation. Our findings also suggest that defined genes, which are specified by retinoids and their receptors, may account for the pleiotropic effect of vitamin A compounds. PMID- 2546160 TI - Transgenic mouse model for human gastric carcinoma. AB - To understand the pathogenesis that may be induced by human adenovirus type 12 (Ad12), we have generated transgenic mice carrying the Ad12 early region 1 under control of the mouse mammary tumor virus long terminal repeat. Eleven of 11 male founder mice, but only 2 of 12 females, died between 3 to 4 mo of age. Death was associated with presence of tumors at or near the squamocolumnar junction of the stomach. Microscopically, these multifocal tumors appeared to arise from hyperplastic epithelium and showed features consistent with adenocarcinoma or adenosquamous carcinoma. High levels of expression of both the Ad12 E1A and E1B genes were seen in the tumor-bearing stomach. Various levels of expression were also detected in other tissues, although the stomach was the only organ with detectable pathology. These observations suggest an organ-specific action of the Ad12 early region 1 gene products. This transgenic mouse model provides an experimental system for studying the development of human carcinomas at sites of transition from squamous to columnar epithelium. PMID- 2546161 TI - Coronavirus subgenomic minus-strand RNAs and the potential for mRNA replicons. AB - The genome of the porcine transmissible gastroenteritis coronavirus is a plus strand, polyadenylylated, infectious RNA molecule of approximately 20 kilobases. During virus replication, seven subgenomic mRNAs are generated by what is thought to be a leader-priming mechanism to form a 3'-coterminal nested set. By using radiolabeled, strand-specific, synthetic oligodeoxynucleotide probes in RNA blot hybridization analyses, we have found a minus-strand counterpart for the genome and for each subgenomic mRNA species in the cytoplasm of infected cells. Subgenomic minus strands were found to be components of double-stranded replicative forms and in numbers that surpass full-length antigenome. We propose that subgenomic mRNA replication, in addition to leader-primed transcription, is a significant mechanism of mRNA synthesis and that it functions to amplify mRNAs. It is a mechanism of amplification that has not been described for any other group of RNA viruses. Subgenomic replicons may also function in a manner similar to genomes of defective interfering viruses to lead to the establishment of persistent infections, a universal property of coronaviruses. PMID- 2546163 TI - Ultrastructural evidence of receptor-mediated endocytosis during early mouse limb morphogenesis: a three dimensional analysis using deep etching and scanning electron microscopy. PMID- 2546164 TI - 3-D changes in neuroblastoma x glioma hybrid (NG 108-15) cell differentiation as studied by SEM and TEM. PMID- 2546162 TI - Synthesis and characterization of a series of diarylguanidines that are noncompetitive N-methyl-D-aspartate receptor antagonists with neuroprotective properties. AB - Four diarylguanidine derivatives were synthesized. These compounds were found to displace, at submicromolar concentrations, 3H-labeled 1-[1-(2 thienyl)cyclohexyl]piperidine and (+)-[3H]MK-801 from phencyclidine receptors in brain membrane preparations. In electrophysiological experiments the diarylguanidines blocked N-methyl-D-aspartate (NMDA)-activated ion channels. These diarylguanidines also protected rat hippocampal neurons in vitro from glutamate-induced cell death. Our results show that some diarylguanidines are noncompetitive antagonists of NMDA receptor-mediated responses and have the neuroprotective property that is commonly associated with blockers of the NMDA receptor-gated cation channel. Diarylguanidines are structurally unrelated to known blockers of NMDA channels and, therefore, represent a new compound series for the development of neuroprotective agents with therapeutic value in patients suffering from stroke, from brain or spinal cord trauma, from hypoglycemia, and possibly from brain ischemia due to heart attack. PMID- 2546165 TI - The arachidonic acid cascade and multistage carcinogenesis in mouse skin. PMID- 2546166 TI - The role of papillomaviruses in human and animal epithelial neoplasia. PMID- 2546167 TI - Interaction of local anesthetics with cytochrome c oxidase. PMID- 2546168 TI - Electrical field effects: their relevance in central neural networks. PMID- 2546169 TI - Porous hydroxyapatite as an onlay bone-graft substitute for maxillofacial surgery. AB - This paper chronicles 3 years of a continuing study comparing porous hydroxyapatite to autogenous bone grafts as onlays in maxillofacial surgery. Twenty-five patients, seen from June of 1984 to May of 1985, underwent onlay augmentation on various maxillary and mandibular locations. A total of 68 onlay augmentation sites comparing Interpore porous hydroxyapatite and autogenous bone were followed for 2 years or more. This long-term study compares these substances in radiologic longevity, histologic incorporation, clinical function, and aesthetic appearance. PMID- 2546170 TI - Repeated exposures intensify rather than diminish the rewarding effects of amphetamine, morphine, and cocaine. AB - It is commonly believed that repeated exposures diminish the pleasurable effects of drugs and hence that pleasure must have only a minor role in addiction. In six experiments with rats, repeated exposures to amphetamine, morphine, or cocaine were found to enhance the drug-induced rewarding effect as measured by conditioned place preference. Thus, sensitization to the rewarding effect, rather than tolerance, was obtained. Also, cross-sensitization was obtained; exposures to amphetamine enhanced the rewarding effect of morphine and vice versa; similarly, exposures to morphine enhanced the rewarding effect of cocaine. These findings support a new theory: drugs of abuse are addictive because repeated exposures sensitize the central reward mechanism so that drug taking produces a progressively greater reinforcing effect each time it occurs. PMID- 2546171 TI - Beta-adrenoceptor binding correlates with behaviour of rats in the open field. AB - Rats were injected IP once daily for 14 consecutive days with propranolol (5 mg/kg), yohimbine (2.5 mg/kg) or saline vehicle. A fourth group was unhandled during this time. Each rat was then placed in an open field for 4 min and its activity and defaecation recorded. Immediately after this, the animals were killed and cerebral cortices removed for radioligand binding to alpha 2- and beta adrenoceptors and measurement of noradrenaline content. We report two sets of findings. First, beta-adrenoceptor density correlated positively, and affinity negatively, with the number of movements towards the centre of the field in the final 3 min of the trial. alpha 2-Adrenoceptor Kd, in contrast, correlated both with movements around the field and those directed towards the centre. Secondly, whereas the only specific drug effect was an increase in defaecation after treatment with propranolol, beta-adrenoceptor density was increased and affinity decreased in all injected groups, suggesting a non-specific effect of the stress of injection. Movements to and from the centre of the field were also increased in injected groups during the first minute of the trial. In both sets of findings the association of beta-adrenoceptor density with greater resistance to stress is hard to reconcile with existing theories of the role of beta-adrenoceptors in behavioural responses to stress. PMID- 2546173 TI - Vasoactive intestinal peptide control of renal adenylate cyclase: in vitro studies of canine renal membranes and cultured canine renal epithelial (MDCK) cells. AB - Vasoactive intestinal peptide (VIP) has been shown to stimulate adenylate cyclase activity in plasma membranes isolated from canine renal cortex, outer and inner medulla in vitro. Though related hormones such as glucagon also stimulate adenylate cyclase in these membrane preparations, it is likely that VIP interacts with specific VIP receptors since the VIP receptor antagonist, (4Cl-D-Phe6, Leu17)-VIP, is capable of reducing the response to VIP, but not that to glucagon. Also binding of 125I-VIP to cortical renal plasma membranes shows competition by unlabelled VIP, but not by glucagon. Strain 1 (and clone CL(8)1b cells derived from the established cultured dog kidney cell line, MDCK, have been shown also to respond selectively to VIP by an increase in adenylate cyclase activity and cyclic AMP accumulation in intact cells. A physiological correlate of VIP activation of adenylate cyclase has been sought by addition of VIP to reconstituted epithelial monolayers of strain 1 MDCK cells clamped in Ussing chambers. VIP addition to the basal-lateral cell aspects generates an inward short-circuit current that is sensitive to replacement of medium Cl- by NO3-, and to inhibition by the Cl- channel blocker, 3-nitro-2(3-phenylpropylamino)-benzoic acid, consistent with VIP stimulation of transepithelial Cl- secretion. PMID- 2546172 TI - Effects of a single or repeated administration of the benzodiazepine inverse agonist FG7142 on behaviour and cortical adrenoceptor binding in the rat. AB - We have reported previously an increase in the number of beta-adrenoceptors in mouse cerebral cortex 7 days after kindling of seizures by repeated once-daily administration of the benzodiazepine receptor inverse agonist, FG7142. In subsequent experiments, an even larger increase in beta-adrenoceptor number was found 7 days after a single injection of this compound. The present experiments investigated whether FG7142-induced changes in adrenoceptor binding are also found in the rat and whether the effects of a single and repeated injections of this drug differ quantitatively. In view of the anxiogenic effects of FG7142, we have also tested for parallel changes in behaviours associated with anxiety and exploration. Nine days after a single injection of FG7142, the number of beta adrenoceptors in the cerebral cortex was greater than that found after repeated administration of this compound; this difference was statistically significant. There was no difference in beta-adrenoceptor binding to tissues from chronically FG7142-treated and vehicle-injected animals and there were no changes in alpha 2 adrenoceptor binding or noradrenaline levels after either a single or repeated FG7142 treatment. Neither single nor repeated FG7142 treatment modified spontaneous behaviour in either the elevated plus-maze test of anxiety or the holeboard test of exploration. The behavioural effects of yohimbine and clenbuterol in these tests were also unaffected by FG7142. We discuss the possibility that the difference in the effects of a single and repeated administration of FG7142 on beta-adrenoceptor binding is related to the expression of kindled seizures. PMID- 2546174 TI - The effect of a cell-permeable diacylglycerol analogue on single Ca2+ (Ba2+) channel currents in the insulin-secreting cell line RINm5F. AB - Depolarization evokes opening of single Ca2+ (Ba2+) channels of the 'L' type in the insulin-secreting cell line RINm5F. Stimulation of the cells with the membrane-permeable diacylglycerol analogue didecanoylglycerol (DC10; 5 micrograms/ml) markedly enhanced the proportion of time the channels spent in the open state during depolarizing voltage pulses. Similar effects have previously been demonstrated with glyceraldehyde stimulation and it therefore seems possible that carbohydrate-evoked cellular Ca2+ uptake is mediated via protein kinase C activation. PMID- 2546175 TI - Prognostic factors for recurrence and cosmesis in 393 patients after radiation therapy for early mammary carcinoma. AB - Between 1978 and 1985, 393 of 2,765 (14%) patients with operable cancer of the breast (clinical stage T0-3N0-2M0) were irradiated after excisional biopsy and staging axillary dissection. Of 77 patients with microscopic axillary metastases, 68 received systemic adjuvant therapy. Treatment failed locally in 26 cases, and there were seven patients with distant metastasis. The three major factors for increased local treatment failure were (a) age below 40 years (P = .003), (b) negative estrogen receptor assay result (P = .03), and (c) failure to deliver a radiation boost dose when tumor was present at the margin of the specimen (P = .002). The size of the tumor, the nodal status, the progesterone receptor assay result, and the presence of ductal carcinoma in situ mixed with infiltrating carcinoma did not show a significant influence on local recurrence. In 274 of 393 (70%) patients, cosmesis was evaluated. The four major factors affecting cosmesis favorably were (a) utilization of a wedge (P less than .0001); (b) treatment of two fields a day (P less than .0001); (c) failure to use a separate treatment port to the regional lymph nodes, so as to avoid field junctions (P = .0003); and (d) small size of specimen (less than 50 cm2) (P = .0171). A second or third cancer was found in 39 of the 393 (10%) patients; contralateral breast cancer was the most common form (n = 23), followed by genitourinary cancer (n = 5). The most frequent complication was arm edema (6%). PMID- 2546176 TI - Circumscribed intraductal carcinoma of the breast. PMID- 2546177 TI - Modulation of second messenger function in rat brain by in vivo alteration of receptor sensitivity: relevance to the mechanism of action of electroconvulsive therapy and antidepressants. AB - 1. The second messengers cyclic AMP and inositol triphosphate are the intracellular mediators for a number of neurotransmitters for which receptors exist on brain neurons. 2. Up- or down-regulation of these receptors in general produce corresponding changes in the associated second messenger systems. 3. Chronic administration of antidepressants including electroconvulsive shock to rats produces a number of changes in cerebral receptors, notably down-regulation of beta-adrenergic and serotonin 5-HT2 receptors and up-regulation of alpha-1 adrenergic receptors. 4. The changes in receptor number induced by such antidepressant treatments are in general accompanied by corresponding changes in the associated second messenger reactions. 5. Antidepressant administration has also been shown to induce increased post-receptor mediated adenylate cyclase activity in cortical membranes, and similar effects have also been reported in striatum after chronic administration of neuroleptics. The relevance of these effects to the mechanism of action of the drugs is discussed. PMID- 2546178 TI - Naloxone binding in the frontal cortex of postmortem human brain. AB - 1. The binding of 3H-Naloxone to opiate receptors was characterized in the frontal cortex of human post-mortem brain samples and age related changes in opiate receptors were investigated. 2. Our study revealed the presence of two binding sites with different affinities for naloxone. 3. With increasing age, the opiate receptors tend to show greater affinity for the agonistic conformation; this may imply a decline in endogenous opioid peptides with age. PMID- 2546179 TI - Pentobarbital-like electroencephalographic rubral rhythm induced by ketamine in rabbits. AB - 1. The effects caused by the dissociative anaesthetic drugs (PCP, KT), the sigmaopiates (SKF 10.047, cyclazocine), and the mixed excitatory amino acid antagonist CCP on the electrical activity of the red nucleus in rabbits were compared with PB. 2. Ketamine (10 mg/kg, i.v.) induced the appearance of a pentobarbital-like EEG synusoidal rhythm characterized by an increase in amplitude and a decrease in frequency of the basal electrical activity at the red nucleus level. 3. Both pentobarbital and ketamine induced rhythms were blocked by the GABA-antagonist pentylenetetrazol at the subconvulsant dose of 10 mg/kg, i.v. 4. Phencyclidine, SKF 10.047, cyclazocine, and the mixed excitatory amino acid antagonist CCP failed to affect the basal electrical activity of the red nucleus. 5. These data indicate an interaction of ketamine on the GABA neurotransmission at the level of cerebello-rubral pathways which the other PCP/sigma opiates did not present. PMID- 2546180 TI - CCK modulation of cerebellar afferents: dopaminergic involvement. AB - 1. Dopamine agonists and dopamine releasers act at the level of the striatum to activate a mossy fiber pathway which in turn increases cerebellar cGMP in the rodent. 2. CCK acts to both decrease basal cerebellar cGMP levels as well as to antagonize increases in cerebellar cGMP elicited by dopaminergic agonists and releasers. 3. This CCK action involves a "central-type" CCK receptor population which resides outside of the cerebellum. 4. These data are consistent with the pharmacology for CCK inhibition of dopamine release in the striatum; suggesting that CCK effects on cerebellar cGMP may be mediated by antidopaminergic actions within this striatum. PMID- 2546181 TI - NMDA-coupled and uncoupled forms of the PCP receptor: preliminary in vivo evidence for PCP receptor subtypes. AB - 1. As reported for many other PCP receptor actions, the pharmacological profile of PCP receptor agonists and NMDA receptor antagonists were similar with regard to their effects on cerebellar cGMP levels in vivo. 2. PCP receptor agonists act to increase mesocortical dopamine (DA) metabolism and release. 3. This receptor action is stereospecific and is both dose- and time-dependent. 4. The actions of PCP on DA metabolism appear to involve PCP receptors both in the ventral tegmental area (VTA) and the cortical nerve terminal regions. 5. In contrast to many other systems which have been studied, competitive NMDA antagonists do not act in a manner similar to PCP agonists, with regard to mesocortical DA metabolism. 6. Sigma receptor ligands and NMDA agonists also do not alter mesocortical DA metabolism. 7. These data suggest that the PCP receptor population which modulates mesocortical dopaminergic neurons is not coupled to NMDA receptors. PMID- 2546182 TI - [Genetics of Caenorhabditis elegans]. PMID- 2546183 TI - [Mechanism of energy conversion in retinal proteins]. PMID- 2546184 TI - [Photo-reaction of bacteriorhodopsin and mechanism of proton pump]. PMID- 2546185 TI - [Structural and functional analogy of eucaryotic rhodopsins and hormone receptors]. PMID- 2546186 TI - [Structural elucidation of phototransduction proteins]. PMID- 2546187 TI - [Control of cGMP level and light-adaptation in vertebrate photoreceptors]. PMID- 2546188 TI - Essential fatty acid deficiency inhibits early but not late leukocyte infiltration in rabbit myocardial infarcts. AB - Essential fatty acid (EFA) deficiency, induced by elimination of the dietary (n 6) fatty acids, has been shown to limit inflammatory cell influx and consequent enhanced eicosanoid production in experimental glomerulonephritis and hydronephrosis. To determine whether EFA-deficiency exerts anti-inflammatory effects following left ventricular myocardial infarction (LVMI), male weanling rabbits were fed EFA-deficient diet for 3 months prior to 60 minutes of distal left circumflex coronary artery occlusion followed by reperfusion. One and 4 days later, corresponding to infiltration of cardiac tissue with polymorphonuclear (PMN) and mononuclear leukocytes respectively, infarcted hearts were buffer perfused and stimulated to produce eicosanoids with f-met-leu-phe or bradykinin. One day following LVMI, the hearts of EFA-deficient rabbits demonstrated a marked suppression of PMN infiltration and eicosanoid production relative to controls. Four days following myocardial infarction, no differences were observed in mononuclear cell invasion, collagen deposition, or eicosanoid production between EFA-deficient and normal hearts. Our data show that EFA-deficiency inhibits PMN influx and consequent enhanced eicosanoid production without affecting the later appearance of mononuclear cells, collagen deposition, or eicosanoid production. Recent studies have shown that suppression of PMN invasion limits the extent of tissue damage following LVMI. Selective inhibition of PMN infiltration is possible and may be useful in the management of acute myocardial infarction. PMID- 2546189 TI - Prostacyclin contributes to inhibition of hypoxic pulmonary vasoconstriction by alkalosis. AB - The mechanism by which extracellular alkalosis inhibits hypoxic pulmonary vasoconstriction is unknown. We investigated whether the inhibition was due to intrapulmonary production of a vasodilator prostaglandin such as prostacyclin (PGI2). Hypoxic vasoconstriction in isolated salt-solution-perfused rat lungs was blunted by both hypocapnic and NaHCO3-induced alkalosis (perfusate pH increased from 7.3 to 7.7). The NaHCO3-induced alkalosis was accompanied by a significant increase in the perfusate level of 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha), an hydrolysis product of PGI2. Meclofenamate, an inhibitor of cyclooxygenase, counteracted both the blunting of hypoxic vasoconstriction and the increased level of 6-keto-PGF1 alpha. In intact anesthetized dogs, hypocapnic alkalosis (blood pH increased from 7.4 to 7.5) blunted hypoxic pulmonary vasoconstriction before but not after administration of meclofenamate. In separate cultures of bovine pulmonary artery endothelial and smooth muscle cells stimulated by bradykinin, the incubation medium levels of 6-keto-PGF1 alpha were increased by both hypocapnic and NaHCO3-induced alkalosis (medium pH increased from 7.4 to 7.7). These results suggest that inhibition of hypoxic pulmonary vasoconstriction by alkalosis is mediated at least partly by PGI2. PMID- 2546190 TI - Regulation of prostaglandin production in cultured gastric mucosal cells. AB - The aims of this study were to investigate whether exogenous prostaglandin modulates prostaglandin biosynthesis by cultured gastric mucosal cells, and to clarify the role of cyclic nucleotides in the possible modulation of prostaglandin production. After pretreatment for 30 min with buffer alone (control) or 1 to 100ng/ml PGE2, cells were incubated with 4 uM arachidonic acid for 30 min. Pretreatments with greater than 5ng/ml PGE2 inhibited arachidonate induced PGE2 and PGI2 production in a dose-dependent fashion, as compared with control, with inhibition by 64 +/- 8% and 75 +/- 4% respectively, at 100ng/ml PGE2. PGE2, at 100ng/ml, significantly increased intracellular cAMP accumulation, but pretreatment with dibutyryl cAMP (0.01-mM) did not alter the amounts of arachidonate-induced PGE2 production. Furthermore, while greater than 10ng/ml PGE2 increased cGMP production dose-dependently, preincubation with dibutyryl cGMP (0.001-0.1mM) also failed to affect PGE2 synthesis significantly. In addition, pretreatment with isobutyl-methyl-xanthine, while increasing accumulation of cellular cyclic nucleotides, did not significantly change PGE2 production. Calcium ionophore A23187-induced PGE2 production was also inhibited by pretreatment with PGE2. These results indicate that exogenous PG inhibits subsequent arachidonate or A23187-induced PG biosynthesis in rat gastric mucosal cells, and suggest the possibility that PG regulates its own biosynthesis via feedback inhibition independent of cyclic nucleotides in these cells. PMID- 2546192 TI - Clinical significance of 99mTc-N-pyridoxyl-5-methyltryptophan (99mTc-PMT) in the diagnosis of intrahepatic masses. AB - Hepatobiliary scintigraphy with 99mTc-N-pyridoxyl-5-methyltryptophan (99mTc-PMT) was carried out on 48 patients with intrahepatic masses, 44 with hepatocellular carcinoma and one each of hepatocellular adenoma, focal nodular hyperplasia, cholangiocellular carcinoma, and adenoid cystic carcinoma. Scans were performed twice, early scan (30 min post i.v.) and delayed scan (2.5 h post i.v.), and the delayed scan was used for assessing the accumulation of 99mTc-PMT in the intrahepatic masses. In the hepatocellular carcinoma group, based on individual patients, 17 out of 44 (38.6%) showed accumulation of 99mTc-PMT in various degrees; and based on individual masses, accumulation was noted in 21 out of 55 masses (38.2%). However, only the cases which had not received transarterial infusion of anti-cancer drugs (TAI) and/or blocking agents (TAE) were taken into consideration, 9 out of 18 patients (50%) and 12 out of 25 masses (48.0%) were found capable of picking up 99mTc-PMT. A case of hepatocellular adenoma showed a strong accumulation of 99mTc-PMT in the mass which was depicted as a defect on the 99mTc-colloid scan and did not show a significant accumulation of 67Ga. In a case of focal nodular hyperplasia, there were two space-occupying lesions (SOLs), one of which showed a clear-cut defect on the 99mTc-colloid scan and the other which showed only a distorted uptake pattern. However, both masses were strongly positive with 99mTc-PMT. 99mTc-PMT scintigraphy is useful in connection with 99mTc-colloid scan and sometimes with 67Ga-citrate in the diagnosis of intrahepatic masses originating from hepatocytes. PMID- 2546193 TI - [Evolution of the echographic picture in primary liver tumor associated with cirrhosis]. AB - To study the sonographic (US) evolution of hepatocellular carcinoma, 53 tumors in 45 untreated patients were observed regularly with real-time US for a period of 6 to 56 months. At the beginning, 25 tumors were hypoechoic, 18 isoechoic, 4 hyperechoic, and 6 had mixed hypo/hyper echo patterns. At the follow-up, 7 initially hypoechoic tumors had changed to hyperechoic or to mixed echo patterns; 8 hypoechoic tumors had become isoechoic; 9 of the 25 initially hypoechoic neoplastic lesions had maintained the same echo density. Ten of the 15 initially isoechoic tumors had changed to mixed echo patterns, and 5 had remained unchanged. Three initially isoechoic lesions and a hypoechoic one had turned into diffuse patterns; 2 initially hyperechoic neoplastic lesions had remained unchanged; 1 had switched into hypoechoic, and 1 changed to mixed echo pattern; 4 out of 6 tumors with echo pattern had remained unchanged, 1 had become hyperechoic and 1 hypoechoic. The current study has proven various tumors less than or equal to 3 cm phi to be isoechoic, and most tumors greater than 3 cm phi to have mixed hypo/hyper echo patterns. The echogenicity of small hepatocellular carcinomas increases with the tumor growth, and remains unchanged when they do not increase in size. PMID- 2546191 TI - Effects of inadequate vitamin E and/or selenium nutrition on the release of arachidonic acid metabolites in rat alveolar macrophages. AB - Effects of vitamin E and/or selenium (Se) deficiency on the secretion of arachidonic acid metabolites by zymosan-stimulated pulmonary alveolar macrophages (AM) were examined using cells from male Long-Evans hooded rats fed torula-yeast based diets with or without the supplementation of vitamin E (150 IU/kg) or Se (0.5 mg/kg). Alveolar macrophages obtained by lavage were purified by adherence and cultured for 4 h in Hank's balanced salt solution containing bovine serum albumin (0.1%) and zymosan (300 micrograms/ml). The arachidonic acid metabolites present in the culture supernatant were measured by radioimmunoassay. Altered vitamin E and Se nutrition had no effect on the number of cells or cell types recovered from the pulmonary airways. Alveolar macrophages derived from animals fed on diets deficient in vitamin E or Se or both nutrients secreted higher levels of prostaglandin E2 and thromboxane B2. Levels of both 5 hydroxyeicosatetraenoic acid and leukotriene B4 were significantly increased only in the group fed the diet adequate in Se but deficient in vitamin E. Our data suggest that vitamin E and Se might play an important role to control the levels of several physiologically and pathologically important arachidonic acid metabolites. PMID- 2546194 TI - [Pulmonary blastoma. A childhood case report and review of the literature]. PMID- 2546195 TI - [Sonographic differential diagnosis of extrahepatic cholestasis]. AB - The value of sonography for aetiological diagnosis was examined in 196 patients with extrahepatic cholestasis. For each patient a diagnosis was prospectively worked out by sonography and compared with the true diagnosis established by endoscopic retrograde pancreaticocholangiography (ERCP), and/or computed tomography (CT), surgical intervention, autopsy and unequivocal clinical course. In 72 patients cholangiolithiasis was the cause of the obstruction. It was diagnosed by sonography with a sensitivity of 86% and a specificity of 93%. With 25 patients with another benign cause of cholestasis the diagnosis was made by sonography in 23 cases. In 93 patients the underlying malignant disease was recognised sonographically with a sensitivity of 95% and a specificity of 100%. PMID- 2546196 TI - [A prototype double coil for MR breast measurement]. AB - The first application of a double coil for routine MR mammography is described: by this method it is possible to examine both breasts simultaneously. This is important in order to halve the time of the examination and to allow comparison of the two breasts during dynamic contrast studies. The reduction in signal-to noise ratio, compared with a single coil and flow artifacts of the heart do not interfere with diagnosis. The axillary region is better shown than with a single coil. Examination of 22 patients with subsequent histological control has shown that the information about signal increase after the injection of Gd-DTPA obtained from single coil can be applied equally to the double coil. PMID- 2546197 TI - [Imaging of a duplication of the small intestine using 99mTc pertechnetate]. PMID- 2546198 TI - [Spontaneous chylothorax: its diagnosis through a combination of lymphography and CT]. PMID- 2546199 TI - [Calcified untreated non-Hodgkin's lymphoma of the mediastinum]. PMID- 2546200 TI - [The growing fracture in children]. PMID- 2546201 TI - Posterior lumbar apophyseal ring fracture simulating a mass lesion in computed tomograms. PMID- 2546202 TI - [Fibromuscular dysplasia in reversible carotid stenosis with temporary hemiparesis]. PMID- 2546204 TI - [Percutaneous implantation of an endovascular Gianturco prosthesis in subclavian vein obstruction]. PMID- 2546203 TI - Spontaneous rupture of an AV aneurysm of the right renal artery. PMID- 2546205 TI - [Chemoembolization of hepatocellular carcinoma--computed tomographic follow-up observation]. AB - The prognosis of unresectable hepatocellular carcinomas is bad. Untreated, survival is 1.6 to 2.5 months. Systemic chemotherapy has little effect and is associated with marked side effects. Tumour cells are supplied almost exclusively by the hepatic artery, whereas normal liver tissue is supplied by the hepatic artery and portal vein. Consequently, interruption of arterial blood flow results in damage to the carcinoma without affecting the rest of the liver. Lipiodol is an oily contrast medium which is taken up selectively by the tumour and can be demonstrated in tumour, in the interstitium and in blood vessels. We used lipiodol emulsified with cisplatin and epirubicin successfully in 22 patients with inoperable liver cell carcinomas. The treatment was well tolerated, apart from some nausea and mild upper abdominal discomfort. Survival time from first diagnosis and the first embolisation was ten months longer than expected survival or that following systemic chemotherapy. PMID- 2546206 TI - [Laryngeal and hypopharyngeal carcinoma--the limits and advantages of sonography]. AB - 29 patients with carcinoma of the larynx and the hypopharynx were evaluated not only for staging cervical lymph nodes but also for classification of the primary tumour. With appropriate technique it is possible to assess tumour infiltration (T4) of surrounding tissue of the larynx at all levels. Tumour classification T1 to T3 is possible in patients with tumour of the epiglottis (regio supraglottis, UICC). The endolaryngeal tumour T1 to T3 (regio glottis and subglottis, UICC) still remains the domain of the microlaryngoscopy. The limits of the method are apparent in hypopharynx carcinomas where in only 50% of the patients the tumour could be assessed. PMID- 2546207 TI - [The anatomy of the pterygopalatine fossa in CT]. AB - The pterygopalatine fossa represents a central space in the depth of the facial skull. Acting as a mediator between neighbouring compartments it is traversed by numerous neurovascular structures. High resolution CT is possible not only to depict all those canals and openings but also the soft tissue contents of the fossa itself can be visualised by CT. This paper presents the radioanatomic prerequisites for assessment of the pterygopalatine fossa that may be affected by a variety of pathologic conditions. PMID- 2546209 TI - [Pulmonary segmental artery angiography in intensive care patients]. AB - The indication for wedge angiograms in intensive-care patients within the scope of X-ray chest examination is outlined. The obligatory wedge angiogram in patients with Swan-Ganz catheter is redundant. 4 indications for this special examination are specified. As a new possibility for differentiating between the ventral or dorsal course of pulmonary vessels, an inverse ratio of arborization angles is outlined for the first time. PMID- 2546208 TI - [High-resolution computed tomography of the thorax in lymphangioleiomyomatosis and tuberous sclerosis]. AB - We present three cases that illustrate the HRCT appearance of pulmonary lymphangioleiomyomatosis and pulmonary involvement in tuberous sclerosis. All patients had multiple cystic lesions diffusely distributed throughout the lungs. The lack of fibrotic changes was a prominent feature in all patients. HRCT correlated more closely than conventional CT with the pathologic findings. PMID- 2546210 TI - [CT-guided biopsy: methods, results and complications]. AB - Percutaneous CT-guided biopsies were carried out in 350 patients; on 291 occasions, 14- or 18-gauge and, on 59 occasions, an 0.95 mm cutting biopsy cannula was employed. Using the 14- and 18-gauge needles, a histological diagnosis was possible in 191 cases, a sensitivity of 82.7%. The 0.95 mm cannulas proved successful in 39 of the 59 punctures, producing a markedly lower sensitivity of 66.2%. The most common cause of an incorrect biopsy with large needles was bad position of the needle, whereas the small needles provided inadequate material. Complications which, however, did not require treatment, occurred in 5 patients (1.4%). PMID- 2546211 TI - [Diagnosis of arterial vascular diseases of the lower extremities with color coded duplex sonography]. AB - Fifty-three patients with chronic arterial disease were examined with colour coded duplex sonography and the results compared with those of arteriography. In those vessels where direct demonstration was possible, there was complete agreement regarding the localisations and severity of lesions. There was some difficulty in evaluating distal high-grade stenoses and occlusions. The results indicate that by means of colour-coded duplex sonography, rapid and accurate evaluation of the vessels in the lower extremities can be achieved. The new method represents a significant improvement in noninvasive diagnosis, particularly as compared with traditional duplex sonography. PMID- 2546212 TI - [Embolization by the wall components after the PTA of pelvic arterial stenoses]. AB - Quality and frequency of embolisation caused by angioplasty of stenoses of iliac arteries were investigated in 50 patients. In 18% emboli were found. They mostly consisted of atheromatous material and were visible by macroscopic view in 10% of the patients. Complications caused by embolisation during angioplasty of iliac artery stenosis however are very rare. PMID- 2546213 TI - [Computed tomography and magnetic resonance tomography of the lower leg in chronic venous insufficiency]. AB - Sixty-two lower limbs in 29 women and six men with chronic venous insufficiency (CVI) of grade II and III were examined by CT and eight of these additionally with MRI. The changes were compared with the findings in 21 patients of similar age, but without venous abnormalities. The most important findings were subcutaneous fibrosis with or without calcification, nonspecific infiltration of the extrafascial spaces and degeneration of the Achilles' tendon. The simultaneous occurrence of these changes can be regarded as typical of advanced CVI. CT was somewhat superior to MRI; its use can be justified as part of posttherapeutic follow-up and for special problems. PMID- 2546214 TI - [Imaging diagnosis of aggressive fibromatosis and the MRT-pathological correlation]. AB - Aggressive fibromatoses (desmoid tumours) tend to grow in an infiltrative and destructive manner without metastases. Computed tomography of aggressive fibromatoses yields no uniform attenuation pattern. The tumours are typically isodense when no contrast medium is used and enhance clearly to hyperdense during infusion of contrast medium. In magnetic resonance tomography (MRT) a high signal (long T2) on T2-weighted pulse sequences as well as an accumulation of i.v. Gd DTPA seems to be the characteristic appearance of aggressive fibromatoses, although different signal intensities can be seen. The MRT histopathologic correlation shows increasing signal intensities on T2-weighted sequences dependent on an increment in cellular content of the tumours. Only histopathological methods can provide a definite diagnosis. PMID- 2546215 TI - [The assessment of stability in spinal fractures]. AB - The assessment of stability is one of the most important problems in the analysis of vertebral trauma. There are numerous indirect signs of instability. The "three column model" provides a concept towards a better understanding of instability. Compared with the conventional complex classifications, the new system based on the "three column model" provides certain advantages. PMID- 2546216 TI - [The tuberositas phalangis distalis of the fingers as enthesis and its normal reactions at the margins]. AB - The ungual tuft of the digits corresponds to an enthesis at which usually numerous reactions take place especially in adults. These reactions are to be classified into formation and reduction of bone. The formation of bone is the dominant reaction and causes diagnostic problems only in irregular shapes. On the contrary, the usual reduction of bone after the preceding formation causes more diagnostic problems because the borderline to pathology is a flowing one. Reductions of bone demand a meticulous observation of possible further osseous changes to record systemic enthesopathic or high-bone turnover situations in their early or minimal stages. PMID- 2546217 TI - [Angular measurements of the hindfoot in CT]. AB - The absence of soft tissue superimposition on bone makes CT greatly superior to conventional radiographs, even using special views, for demonstrating the hind foot. Views derived from both methods are compared. Semicoronary and transverse planes have been defined in relation to specific points in the tarsus. In the semicoronary plane, the calcaneus-valgus angle varies from +10 degrees (valgus) to -10 degrees (varus) and the sustentaculum angle varies from 18 to 28 degrees. In the transverse plane, the plantar talocalcaneal angle is 60 degrees to 70 degrees and the calcaneal-cuboidal angle is 20 degrees to 35 degrees. These values are based on measurements of 62 normal feet. They are valuable for judging quantitatively posttraumatic deformities, displacement of fragments and operative results. PMID- 2546218 TI - [Digital subtraction angiography of the hand]. AB - The report deals with 66 digital subtraction angiograms of the hand performed on 61 patients; 60 were carried out by direct puncture of the brachial artery and six by trans-femoral catheterisation. In 39 patients a peripheral organic vascular abnormality was demonstrated (usually arterial sclerosis, thromboangiitis obliterans or collagenosis). Purely functional flow abnormalities were found in 17 patients. In 15 patients an organic abnormality was demonstrated, but its type could not be identified. Intra-arterial DSA is able to differentiate between functional abnormalities of blood flow and peripheral organic vascular abnormalities. PMID- 2546219 TI - [ROC: a method for comparing the diagnostic performance of imaging procedures]. AB - ROC analysis permits an evaluation of the diagnostic performance of imaging systems by incorporating the individual decision threshold of the diagnostician that determines the relation between sensitivity and specificity of a tested system. It therefore facilitates the comparison between different imaging systems and observers. A thorough knowledge and sophistication of this methodology is of special importance in the comparison of digital and conventional projection radiography because the images in question may have a very similar character. The method is explained and special points of interest (standard of "truth", size of sample, lesion conspicuity and localisation, viewing time and determination of significance) are emphasised. PMID- 2546220 TI - [Size and density determination by thin-layer CT in relation to structural diameter and the contrast of the surrounding tissue]. AB - The reliability of CT size and density measurements has been examined by using phantom measurements with constant examination and reconstruction parameters (Somatom DRH/Siemens). Ten plexiglass rods with diameters from 2 to 15 mm. were measured singly in (1) air, (2) isodense (146 Hu) and (3) hyperdense medium (985 Hu) using 2 mm. sections. In the isodense medium there was only small variation in density measurements (124 to 131 Hu). Measurements of size showed a maximum error up to -35%. Density measurements in the hyperdense medium produced raised values (197 +/- 17 Hu at 15 mm.). There was an even greater increase for rods smaller than 5 mm. Measurement of size was correct, as it was in air (maximum deviation 8%). Density measurements in air for diameters less than 6 mm. were significantly reduced. PMID- 2546221 TI - [Experimental determination of weighting functions in magnetic resonance tomography]. AB - Weighting functions are defined and calculated for the pulse sequences spin-echo and inversion recovery. These weighting functions make possible a qualitative discussion of the term "weighting" in MR imaging. Furthermore, a new method has been developed to determine weighting functions experimentally. A comparison between the results obtained and the theoretical predictions shows a sufficiently good agreement. This method can also be applied with other pulse sequences. PMID- 2546222 TI - Addition of reducing agents to the peroxidase-o-phenylenediamine buffer reduces background of enzyme immunoassays. AB - The concentrations of o-phenylenediamine (OPD), H2O2, citrate and H+ in a substrate buffer for peroxidase immunoassays were optimized for minimal background. The background was reduced 2-3 fold with 5.5 mM OPD, 3 mM H2O2, 150 mM citric acid/sodium citrate, pH 4.8, and the reproducibility interassay was increased. A further 3-5 fold reduction of the background was obtained by the addition of 1.5 mM acetanilide, 0.14 mM beta-mercaptoethanol and 5 mM nitrilotriacetic acid to the substrate buffer. This low-background substrate buffer allows increased sensitivity and lowers the interassay variation coefficient. It has been used successfully in peroxidase immunoassays of human C reactive protein, human antiestreptolysin and human rheumatoid factor. PMID- 2546223 TI - Effect of 3-(2,2,2-trimethylhydrazinium) propionate, gamma-butyrobetaine hydroxylase inhibitor, on isoproterenol-induced mitochondrial dysfunction. AB - The present study was designed to investigate whether or not reduction of carnitine content could protect isoproterenol (ISP)-induced myocardial injury using 3-(2,2,2-trimethylhydrazinium) propionate (TMHP), gamma-butyrobetaine hydroxylase inhibitor. Rats were divided into 4 groups; the control group: untreated, the TMHP-1 group: TMHP (100 mg/kg) was administered intraperitoneally, and ISP (10 mg/kg) was administered subcutaneously on the following day, the TMHP 7 group: TMHP (100 mg/kg) for 7 successive days, and ISP (10 mg/kg) on the eighth day, the ISP group: ISP (10 mg/kg) was administered. Rats were cervically dislocated 15 hours after ISP administration, and heart mitochondrial electron transport activity (NADH-cytochrome c reductase, succinate-cytochrome c reductase, and cytochrome c oxidase) were measured enzymatically. Activity of succinate-cytochrome c reductase was not affected significantly by ISP, however, NADH-cytochrome c reductase and cytochrome c oxidase were significantly reduced in the ISP and TMHP groups. Administration with TMHP for 7 successive days lessened the reduction of the activities. Mitochondrial electron-transport system plays an important role in cellular energy transduction. These results suggested that mitochondrial dysfunction induced by ISP is related to carnitine-dependent fatty acid metabolism and that TMHP reduces the myocardial injury. PMID- 2546224 TI - A comparative study of the effects of ketoconazole and fluconazole on 17-beta estradiol production by rat ovaries in vitro. AB - In this study we have compared the effects of ketoconazole and fluconazole, a novel triazole antifungal agent, on 17-beta estradiol production in rat ovaries in vitro. For both compounds there was a lag phase, immediately after addition to the test system, during which the rate of oestradiol synthesis remained at control values. This may have been due to the time required for uptake of the compound and transfer to its site of action or for depletion of endogenous pools of intermediates. After the lag phase both compounds produced a reduction in the rate of estradiol synthesis. At any given concentration, fluconazole produced a reduction which was substantially less than that observed with ketoconazole. Indeed 2 microM ketoconazole reduced the rate of oestradiol production by greater than 90% while 10 microM fluconazole caused only a 70% reduction. These findings are consistent with reports that these compounds are inhibitors of cytochrome P450 and with the reduced sensitivity of mammalian cytochrome P450 to fluconazole as compared with ketoconazole. PMID- 2546226 TI - The role of thyroid hormones in the control of beta-adrenergic receptors in rat mammary gland. AB - We have studied the effect of hyperthyroidism and hypothyroidism on the lactating rat mammary gland number of beta-adrenergic receptors and basal intracellular cyclic AMP levels. We also describe the effects of isoproterenol, cholera toxin and forskolin challenge on these parameters. Using cyclic AMP levels as an indication of receptor functionality our results indicate that both thyroid states are accompanied by a decrease in the response to beta-adrenergic receptor stimulation. PMID- 2546225 TI - Pharmacokinetics of [3H]-buprenorphine in the rat. AB - The present study was undertaken to evaluate the potential usefulness of 11C buprenorphine (bup) as a ligand for investigating opioid receptors in living primates, including humans, using positron-emission transaxial tomography (PETT). Because PETT studies of receptor function are best carried out under conditions of low receptor occupancy, the pharmacokinetics, uptake into brain, and specific binding to opioid receptors within brain of 3H-bup were examined in rats under conditions in which occupancy of opioid receptors by 3H-bup never exceeded 2% of sites in the brain at any time point examined. Male Sprague Dawley rats (weight range 140-220 grams) were injected s.c. with either naloxone (10 mg/kg) or saline. Five min later, a saline solution containing [15, 16-3H] bup (39 Ci/mmole) was injected into tail veins at a dose of 0.4 microgram/kg body weight. At least 90-95% of radioactivity was cleared from the blood in the first 5 min. In saline pretreated rats, total brain uptake 15 min after injection of 3H-bup was about 0.4% of the administered dose. Ligand specifically bound to receptors may be estimated by comparing the amount of radioactivity in the brain following injection of labeled ligand alone to that obtained when a high concentration of an unlabeled competitor is pre- or co-administered. In the present study, average levels obtained in brain (excluding cerebellum) were higher in saline pretreated rats than in naloxone pretreated rats at all time points and the difference increased with time indicating specific binding to opioid receptors. Specific binding may also be estimated by comparing radioactivity accumulated in brain areas rich in opioid receptors with "background" levels achieved in areas known to be low in opioid receptors, e.g., the cerebellum in rats. In the present study, ratios of the amount of radioactivity in brain (excluding cerebellum) to the amount in the cerebellum increased with time (to about 4 after 60 min) in saline pretreated rats, but remained close to 1 in naloxone pretreated rats. The effects of biological variation were less when specific binding was estimated by the latter method since each animal served as its own control. Tissue distribution of radioactivity to other tissues (blood, skin, muscle, fat, liver, kidney) was similar in naloxone and saline pretreated rats. The results presented here suggest that 11C-bup or an 18F-labeled fluorinated derivative would be a useful ligand for PETT studies. PMID- 2546227 TI - Effect of cardiac arrest time on cortical cerebral blood flow during subsequent standard external cardiopulmonary resuscitation in rabbits. AB - Standard external cardiopulmonary resuscitation (SECPR) produces high cerebral venous and intracranial pressure peaks, low cerebral perfusion pressure, and low cerebral blood flow (CBF). Cerebral viability seems to require 20% of normal CBF, which SECPR cannot reliably generate. We tested the hypothesis that SECPR can produce adequate CBF if started immediately, but not if started after a long period of cardiac arrest (no flow, stasis). Cardiac arrest times of 1, 3, 5, 7 and 9 min were studied in rabbits. We measured unifocal cortical CBF with H2 clearance curves after saturation with H2 10%, O2 50% and N2O 40% by intermittent positive-pressure ventilation (IPPV). Measurements were made during spontaneous circulation (control condition), and then after resaturation immediately before induction of asystole by KCl i.v., and H2 clearance starting at end of arrest time during SECPR-basic life support with IPPV 100% and manual chest compressions (120/min) during asystole. Control cortical CBF was 30-40 ml/100 g brain per min. During asystole and SECPR, CBF greater than 20% normal was achieved only after no flow of 1 min. After longer arrest (no-flow) times, CBF was less than 20% normal. Values were near zero after 7 and 9 min of cardiac arrest. Decrease in mean arterial pressures (MAP) produced by SECPR during asystole paralleled CBF values. Thus, the longer the preceding period of stasis, the lower the MAP and CBF generated by SECPR without epinephrine. This effect may be the result of anoxia induced vasoparalysis and stasis-induced increased blood viscosity. PMID- 2546228 TI - Transcutaneous oxygen tension measurements during hemorrhagic hypoperfusion using Trendelenburg and the pneumatic antishock garment. AB - Transcutaneous oxygen tension (PtCO2) was observed during hemorrhagic hypoperfusion using four therapeutic modalities: pneumatic antishock garment (PASG), 20 degrees Trendelenburg positioning, combined PASG-Trendelenburg, and whole blood infusion. Anesthetized mongrel dogs were mechanically ventilated. A heated transcutaneous oxygen sensor was applied to the skin overlying the sternum. Animals were bled over 10 min of 25% of their calculated blood volume. A therapeutic intervention was applied at the onset of hemorrhage (PASG, Trendelenburg, PASG-Trendelenburg or control). All animals were observed for 20 min, then during a 10-min shed blood reinfusion period, and for 20 min thereafter. PtCO2 was measured continuously and the following were measured serially: cardiac output, mean arterial pressure (MAP), mixed venous oxygen tension (MvO2), and arterial oxygen tension (PaO2). Cardiac index (CI) and the oxygen extraction ratio were calculated. PtCO2 decreased immediately after hemorrhage in all animals. Control values remained consistently below values for active interventions during this time. All groups regained baseline levels of PtCO2 after reinfusion of shed blood volume. PaO2 remained nearly constant during all experiments. MAP and CI fell in all groups following hemorrhage but did so less precipitously in the PASG group. The PASG and PASG-Trendelenburg groups showed the greatest increase in CI during reinfusion. These results suggest that when PtCO2 is relied upon as an indicator of adequacy of resuscitation during moderate hemorrhagic shock, that cutaneous perfusion may be improved by the PASG or Trendelenburg position, and that perfusion is most effectively restored by blood infusion. PMID- 2546229 TI - Algorithm use in the treatment of pre-hospital ventricular fibrillation: an analysis of 160 cases. AB - In a series of 160 consecutive patients with pre-hospital ventricular fibrillation, outcomes were improved if base-station personnel precisely followed the initial 7 steps of standard VF algorithms. This improvement reached statistical significance regarding survival to hospital discharge (17% vs. 6%, P less than 0.05), and reflected a very strong trend with regard to initial resuscitation and admission to hospital (31% vs. 18%, 0.05 less than P less than 0.10). These measures of outcome were even more strongly related to field time, such that patients treated in the field for less than 15 min did far better than patients treated longer. While concordance with algorithms did not independently diminish field time in this study, this probably reflects the fact that paramedics had to establish base hospital contact and receive orders from base personnel in all patients; thus it is probable that allowing paramedics to treat patients in VF, using precise protocols, without prior communication with a base hospital, would diminish field time, and this might lead to even further improvement in patient outcome. PMID- 2546230 TI - Comparisons of French and U.S.A. pediatric intensive care units. AB - Consecutive admissions to two pediatric intensive care units (PICUs) in France (n = 93) and the United States (n = 248) were compared using admission demographics, and daily therapeutic and severity of illness data. Analysis of the major demographic characteristics revealed that patients in the French PICU were younger (median age; 3 months vs. 31 months, P less than 0.001), and more commonly admitted for emergency reasons (92% vs. 66%, P less than 0.05). General resource utilization was similar in both units. However, important differences in the incidences of use of individual monitoring and therapeutic modalities were present. The United States PICU had higher incidences of invasive monitoring modalities (arterial catheters, 66% vs. 4%, P less than 0.001; central venous catheters, 38% vs. 11%, P less than 0.001; pulmonary artery catheters, 8% vs. 1%, P less than 0.01), while the French PICU had higher incidences of labor-intensive monitoring modalities (strict input/output, 75% vs. 47%, P less than 0.0001; greater than 3 stat blood studies/shift, 69% vs. 45%, P less than 0.0001). Patients in France were more likely to receive mechanical ventilation (81% vs. 56%, P less than 0.0001) and nutritional support (40% vs. 7%, P less than 0.05). Mortality rates in both PICUs were similar and accurately predicted by admission day severity of illness scores. We conclude that differential resource utilization, possibly arising from different care philosophies, may result in equivalent care. PMID- 2546231 TI - The effect of cardiopulmonary bypass resuscitation on cardiac arrest induced lactic acidosis in dogs. AB - The adequacy of end organ blood flow following a cardiac arrest varies depending on the artificial reperfusion technique utilized and may critically affect patient outcome. Both oxygen consumption (VO2) and arterial lactate values have previously been used to assess tissue perfusion. Cardiopulmonary bypass resuscitation (CPB) is a reperfusion technique capable of providing near normal end organ blood flow. The purpose of this investigation was to study the effect of femoro-femoral veno-arterial CPB resuscitation compared to standard CPR on VO2 and arterial lactic acid values after a prolonged cardiac arrest. Ten mongrel dogs were electrically fibrillated and left in cardiopulmonary arrest without therapy for 12 min. Resuscitation was attempted according to a standardized protocol utilizing either CPB (n = 5) or standard external CPR (n = 5). Oxygen consumption values and arterial lactic acid samples were obtained at baseline, at timed intervals throughout resuscitation and after return of spontaneous circulation in successfully resuscitated dogs. Baseline hemodynamic and biochemical measurements were similar in both treatment groups (P greater than 0.05). Oxygen consumption (440 +/- 50 ml/min/M2) and mean arterial lactic acid values (7.44 +/- 2.25 mmol/l) were significantly higher at 1 min of resuscitation in CPB-treated dogs compared to dogs treated with CPR (60 +/- 10 ml/min/M2) (3.16 +/- 0.69 mmol/l) respectively (P less than 0.05). Mean arterial lactic acid values rose significantly at each sampling interval during CPR (P less than 0.05) but began to decrease after 5 min of resuscitation in the CPB animals and were not significantly different than baseline after 60 min of bypass (P greater than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546232 TI - Interventions in the therapy of electromechanical dissociation. AB - Electromechanical dissociation (EMD) is a major arrest rhythm for which there is often inadequate treatment. The purpose of this study was to evaluate the different pharmacological and non-pharmacological interventions considered in the treatment of EMD. During the 6-year period, January 1st, 1980 to December 31st, 1985, 503 evaluable adult patients presented in a non-traumatic, non-poisoning cardiopulmonary arrest with the initial rhythm of EMD. One hundred nineteen patients obtained a pulse during resuscitation efforts following drug administration. The average time to obtaining pulses after the last drug administration was 1.97 +/- 2.21 min. The following drugs were last administered prior to transient pulses: bicarbonate, 31/119 (26%); epinephrine, 26/119 (22%); atropine, 26/119 (22%); dopamine, 13/119 (11%); calcium, 11/119 (9%); isoproterenol, 7/119 (6%); other drugs, 5/119 (4%). Ninety-five percent of the successful resuscitations received eight or less drug interventions and all saves received three or less drug interventions. Two hundred twenty-four patients (44.5%) had 288 non-pharmacological interventions. Twenty-three patients developed a pulse after intervention in the following distribution: MAST suit (N = 9), pericardiocentesis (N = 6), fluid challenge (N = 5), needle thoracostomy (N = 1), and intervention combinations (N = 2). The time interval between intervention and the onset of pulse was as follows: MAST suit, 4 +/- 2.8 min; pericardiocentesis, 3.7 +/- 3.6 min; fluid challenge, 4.8 +/- 4.1 min; needle thoracostomy, 6 min. The overall save rate for intervention patients was 0.9% whereas for those not having intervention it was 7.2% (P less than or equal to 0.0003).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546233 TI - Electromechanical dissociation: six years prehospital experience. AB - Electromechanical dissociation (EMD) is the presenting rhythm in approximately 17% of all prehospital cardiorespiratory arrests. Yet, we know comparatively little about the demographic profile of these patients. The purpose of this study was to review historical and resuscitative parameters to help create a demographic profile. For a 6-year period of time from January 1st, 1980 to December 31st, 1985, 503 adult patients presented to a prehospital system in non traumatic, nonpoisoned, cardiorespiratory arrest with an initial rhythm of electromechanical dissociation. The overall average response time was 6.1 +/- 3.2 min. Sixty percent of the patients were witnessed arrests and 65% had bystander initiated CPR. Forty-six percent of the patients had a cardiac history: myocardial infarction 13%, CHF 11% and other 21%. Other pertinent past medical history included diabetes 15%, COPD 10% and seizures 3%. The average age was 69.8 +/- 13.7 years. Fifty-seven percent were male. Forty-three percent were on cardiac medication including: digoxin, 24%; nitroglycerin, 12%; potassium supplements, 9%; propranolol, 8%; isordil, 6%; quinidine, 3%; nitropaste, 3%; and other cardiac medications, 15%. One hundred forty-eight (29%) patients developed a pulse at some time during resuscitative efforts, of these 17 (3.4%) patients responded with a pulse immediately after intubation. The mean time of resuscitation to sustaining pulse was 20 +/- 11 min and the mean resuscitation time to sustaining pressure was 22 +/- 11 min. Nineteen percent were successfully resuscitated, defined as a conveyance of a patient with a pulse and a rhythm to an emergency department. Four point four percent were saved, defined as a patient discharged alive from the hospital. Approximately 53% of the successfully resuscitated patients and 45% of the save patients were determined to have a probable respiratory event as the primary etiology of their arrest. This study attempts to provide some insight into the demographic profile of the patients in EMD. PMID- 2546234 TI - Electrocardiographic characteristics in EMD. AB - Little has been written concerning the initial electrocardiographic (EKG) characteristics and/or changes which occur as the result of treatment in the electromechanical dissociation (EMD) patient. The purpose of this retrospective study was to determine predictive indicators of successful resuscitation in EMD by evaluating various EKG parameters. During 72 months, ending December 31st, 1985, there were 503 non-poisoned, prehospital adult cardiac arrest patients whose initial rhythm was EMD. All patients had their initial prehospital EKG rhythm strip evaluated for rhythm type, rate, the presence of P waves, QT interval and QRS interval. In successfully resuscitated patients, the prehospital initial rhythm analysis and the rhythm analysis on emergency department presentation were compared. Successfully resuscitated patients presenting with EMD had significantly faster initial rates, higher incidences of P waves and average QRS and QT intervals shorter than patients not responding to therapy. Furthermore, successfully resuscitated patients had significantly increased heart rates, developed new onset of P waves, and shortened QT intervals in response to treatment. Successfully resuscitated and save patients had average initial and final QRS complex lengths within normal limits. Organized atrial activity on the initial EKG was also correlated with successful resuscitation. No patient with an initial EKG rhythm of second or third degree AV block survived to hospital discharge. No patient who presented to the emergency department with atrial fibrillation survived to hospital discharge. Similarly, supraventricular tachycaydia following resuscitative efforts appeared to be associated with a negative outcome. Rate normalization following treatment was correlated with save rate. Wide complex rhythms without atrial activity were most highly associated with unsuccessful resuscitation. We believe these observed electrocardiographic characteristics and/or changes in response to treatment may have predictive value in evaluating patients with EMD. PMID- 2546235 TI - Defining electromechanical dissociation: morphologic presentation. AB - Electromechanical dissociation (EMD) is inconsistently defined in the literature. Our definition is the presence of discernible electrical complexes (excluding ventricular tachycardia and ventricular fibrillation) and the absence of palpable pulses. It has been noted that EMD may present with a variety of morphological complexes. It was the purpose of this study to categorize the electrical morphologic characteristics of patients presenting in EMD and to correlate morphology with patient outcome and response to therapy. From the 6-year period, January 1st, 1980 to December 31st, 1985, 503 evaluable adult patients presented to an urban paramedic system in non-traumatic, non-poisoned, cardiorespiratory arrest and were determined to be in EMD. The rhythm strips obtained from paramedics on all patients were retrospectively reviewed and were arbitrarily categorized in the following manner: Group 1, normal QRS width, isoelectric ST and normal appearing T-waves; Group 2A, atrial activity, widened QRS width (greater than or equal to 0.12 ms) or abnormal ST and/or T-waves (ST depression, elevation, slurring or T-wave inversion); Group 2B, same as Group 2A but without atrial activity; Group 3, essentially monophasic, slurred RST complexes. The respective initial distribution was Group 1, 147 (29%); Group 2A, 248 (49%); Group 2B, 60 (12%); Group 3, 48 (10%). The relative frequency of morphologies preceding the attainment of a pulse was as follows: Group 1, 30 (24%); Group 2A, 82 (65%); Group 2B, 8 (6%); Group 3, 6 (5%) (P less than or equal to 0.01 with no significant difference between Group 2B and 3).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546236 TI - Pulmonary infection in human immunodeficiency disease: viral pulmonary infections. AB - Viral pneumonitides are among the known pulmonary complications of human immunodeficiency virus (HIV) infection. Cytomegalovirus (CMV) pneumonitis is the most frequently recognized viral infection involving the lung. Although CMV may occasionally be the sole pathogen found to be responsible for severe pneumonitis in patients with the acquired immunodeficiency syndrome (AIDS), in most cases, its role in causing pulmonary disease is less clear, primarily because of the propensity to infect with a variety of other copathogens. CMV pneumonitis has been difficult to diagnose during life, although techniques utilizing in situ DNA hybridization or monoclonal antibodies for detection of the virus may improve the diagnostic yield of less invasive procedures such as bronchoalveolar lavage. Pneumonitis due to herpes simplex virus, varicella-zoster, and respiratory syncytial virus have occasionally been reported in AIDS patients, and are of practical importance because of the availability of effective treatment. The role of influenza and adenoviruses in causing HIV-related pulmonary complications is unknown, but could be of importance during outbreaks of these infections. Finally, data from several studies now suggest that Epstein-Barr virus or HIV itself or both have a role in the pneumonitis. Further study in this area could provide information leading to more effective management of this common complication of childhood AIDS. PMID- 2546237 TI - WHO CARDIAC Study--its experimental background and progress report. PMID- 2546238 TI - Regression models for time to seroconversion following experimental bovine leukaemia virus infection. AB - This paper develops a parametric model for time to seroconversion after experimental bovine leukaemia virus (BLV) infection, and examines the effects of inoculation route, volume of inoculum, type of inoculation material, and antigen status of donor on seroconversion time. We used parametric and nonparametric statistical methodology to analyse interval data on 150 animals from 13 published reports. The log-logistic model fitted the observed times to seroconversion better than the log-normal or Weibull models, which were the considered alternatives. PMID- 2546239 TI - [Peripheral neuropathies caused by drugs]. AB - Peripheral neuropathy is a common manifestation of chemotherapeutic agents. Most drugs produce a distal axonal degeneration and long axons are predominantly affected, giving a clinical picture characterized by a symmetrical sensory neuropathy. The toxicity of therapeutic agents to the peripheral nervous system particularly includes those used in cancer-chemotherapy (cisplatin, vinca alkaloids), but antimicrobial agents such as isoniazide or nitrofurantoin and vitamin abuse (pyridoxine) are also discussed. PMID- 2546240 TI - [Possible trophic role on the neuromuscular junction of a neuropeptide co existing with acetylcholine in motor neurons of the spinal cord]. AB - The Calcitonin-Gene Related Peptide (CGRP), a neuropeptide present in chick spinal cord motoneurons, increases the levels of surface acetylcholine receptor (AChR) and of the AChR alpha-subunit mRNA in cultured chick myotubes. Cholera toxin (CT), an activator of adenylate cyclase, produces a similar effect which does not add up with that of CGRP. Consistent with this observation, CGRP increases the content of cyclic AMP in chick muscle cells in culture. Tetrodotoxin (TTX), a blocker of voltage-sensitive Na+ channels, elevates the levels of AChR and of AChR alpha-subunit mRNA. This effect is additive with that of CGRP or CT. TPA (12-O-tetradecanoyl phorbol-13-acetate), an activator of protein kinase C, decreases the level of AChR but has no effect on the level of AChR alpha-subunit mRNA. Interestingly, TPA inhibits the increase of AChR alpha subunit mRNA caused by TTX without affecting that produced by CGRP or CT. These data suggest that CGRP, which coexists with acetylcholine in spinal cord motoneurons, could be one of the anterograde factors (or a model of such factor) responsible for the enhanced expression of the genes coding for AChR subunits in subneural nuclei, via the activation of adenylate cyclase. Muscle electrical activity would then inhibit the expression of the same genes in extrajunctional nuclei, via another intracellular pathway. PMID- 2546241 TI - [Unilateral lesions of basal ganglia nuclei in children during myxovirus para influenzae infection]. AB - Unilateral disorders of the basal ganglia in childhood have received little attention. Infections are a rare cause where as infarction is the most common one. We report the case of a 11 year-old boy who presented an acute hemiplegia with hypodense areas limited to the left lenticular, caudate nuclei and internal capsule. Virologic investigations showed a recent infection due to myxovirus para influenzae. Angiography demonstrated no focal vascular disease. The course was favorable with full recovery. PMID- 2546242 TI - Epidemiology of genital papillomaviruses and cervical cancer. AB - Cervical cancer is an extremely common disease. Its natural history has been well described, and individual risk factors have been defined. It is clear from the epidemiologic evidence that cervical cancer has a multifactorial etiology involving infection with sexually transmitted agents such as genital papillomaviruses and cofactors such as pregnancy, smoking, use of hormonal contraceptives, and diet. The evidence implicating papillomavirus as an etiologic agent of cervical cancer has come from a variety of observational laboratory studies. Genital papillomaviruses induce dysplastic lesions. Most invasive cervical cancers contain papillomavirus DNA, as do cell lines derived from cervical cancers. Viral DNA, appears to be integrated into cellular DNA, and integration involves highly conserved, transcriptionally active regions of the viral DNA. PMID- 2546243 TI - Postpartum uterine infection with Clostridium perfringens. AB - Clostridium perfringens is commonly present in the female genital tract. Uterine infection with this organism is a potentially fatal disease infrequently seen in obstetric practice. The manifestations of C. perfringens uterine infection are variable, ranging from endometritis to gas gangrene with fulminant septicemia. The usual precipitating event has been septic abortion, but such infections can also occur spontaneously in uterine tumors and after complicated deliveries requiring mechanical intervention. Diagnosis may be aided by radiologic techniques, and treatment involves high-dose penicillin and possibly surgery. We report two cases and review the clinical presentation and the diagnostic and therapeutic aspects of this disease. PMID- 2546244 TI - Mucocutaneous dissemination of acyclovir-resistant herpes simplex virus in a patient with AIDS. AB - Infections caused by herpes simplex virus (HSV) are a significant source of morbidity in immunocompromised patients. Acyclovir is often used prophylactically and therapeutically in patients with human immunodeficiency virus infection. The emergence of acyclovir-resistant strains of HSV capable of causing disease has been recognized. We report a case in which a thymidine kinase-deficient mutant of HSV caused extensive disease in a patient with AIDS. This case emphasizes that virus recovered from nonhealing lesions should be submitted for further study, which may advance our understanding of the interaction between host defense and drug-resistant strains. PMID- 2546245 TI - Activation of the immune coagulation system by murine hepatitis virus strain 3. AB - Viral infections result in alterations in hemostasis and coagulation. It has previously been shown that susceptibility to murine hepatitis virus strain 3 (MHV 3), a coronavirus, correlates directly with the spontaneous, T lymphocyte instructed expression of a procoagulant monokine that exhibits prothrombin cleaving activity (procoagulant activity [PCA]). A biologic role for PCA in the pathogenesis of MHV-3 infection is suggested by results of in vivo microscopic observations made during acute MHV-3 infection. Recently, it has been demonstrated that prostaglandin E2 abrogates the induction of PCA by MHV-3 both in vivo and in vitro and prevents hepatic necrosis and the associated microcirculatory changes. These data suggest that MHV-3 induces cellular injury through the activation of the coagulation cascade and provide further evidence for a role for PCA in the pathogenesis of MHV-3 infection. PMID- 2546246 TI - Infection and coding strategies of arenaviruses, phleboviruses, and nairoviruses. AB - The infection and coding strategies of three groups of negative-stranded RNA viruses (arena viruses, phleboviruses, and nairoviruses) that include the etiologic agents of hemorrhagic disease in humans have been studied. Arenaviruses have two viral RNA species. The smaller RNA species (S) codes for the viral nucleoprotein (N protein) and for the viral glycoprotein species (G1 and G2, which are derived from a precursor glycoprotein, GPC). The S RNA has an ambisense arrangement. The proteins are translated from subgenomic mRNA species (viz., N protein from a viral-complementary mRNA and glycoprotein from a viral-sense mRNA). The larger arenavirus RNA species (L) is presumed to code for the viral transcriptase/replicase. Phleboviruses and nairoviruses are members of the Bunyaviridae. They both have three species of viral RNA. Other than the sizes of the viral proteins and the viral RNA species, virtually nothing is known about the coding strategy of nairoviruses. Phleboviruses have an ambisense coding arrangement to their smallest (S) RNA species. This S RNA codes for the viral N protein (translated from a viral-complementary mRNA) and a nonstructural protein (translated from a viral-sense mRNA). The middle-size (M) RNA of phleboviruses codes for a precursor to the viral glycoproteins (translated from a viral complementary mRNA). The largest viral RNA (L) is presumed to code for the viral transcriptase/replicase. PMID- 2546247 TI - Epidemiology of hemorrhagic fever viruses. AB - Twelve distinct viruses associated with hemorrhagic fever in humans are classified among four families: Arenaviridae, which includes Lassa, Junin, and Machupo viruses; Bunyaviridae, which includes Rift Valley fever, Crimean-Congo hemorrhagic fever, and Hantaan viruses; Filoviridae, which includes Marburg and Ebola viruses; and Flaviviridae, which includes yellow fever, dengue, Kyasanur Forest disease, and Omsk viruses. Most hemorrhagic fever viruses are zoonoses, with the possible exception of the four dengue viruses, which may continually circulate among humans. Hemorrhagic fever viruses are found in both temperate and tropical habitats and generally infect both sexes and all ages, although the age and sex of those infected are frequently influenced by the possibility of occupational exposure. Transmission to humans is frequently by bite of an infected tick or mosquito or via aerosol from infected rodent hosts. Aerosol and nosocomial transmission are especially important with Lassa, Junin, Machupo, Crimean-Congo hemorrhagic fever, Marburg, and Ebola viruses. Seasonality of hemorrhagic fever among humans is influenced for the most part by the dynamics of infected arthropod or vertebrate hosts. Mammals, especially rodents, appear to be important natural hosts for many hemorrhagic fever viruses. The transmission cycle for each hemorrhagic fever virus is distinct and is dependent upon the characteristics of the primary vector species and the possibility for its contact with humans. PMID- 2546248 TI - Prospects for treatment of viral hemorrhagic fevers with ribavirin, a broad spectrum antiviral drug. AB - Ribavirin, a broad-spectrum antiviral drug, is active against hemorrhagic fever viruses (with the exception of Ebola virus) in cell culture systems. In model infections with arenaviruses in guinea pigs and monkeys, ribavirin has demonstrated both prophylactic and therapeutic efficacy. In therapeutic studies it has not prevented late-onset neurologic disease. In human cases of Lassa fever, it significantly reduces mortality when administered before day 7 of illness to persons at high risk. In rodents and monkeys infected with Rift Valley fever virus, ribavirin therapy resulted in reduced mortality; prophylactic administration to volunteers infected with sandfly fever virus, Sicilian strain, prevented development of illness. Ribavirin increased the number of survivors and the mean time to death in suckling mice infected with Crimean-Congo hemorrhagic fever virus and in suckling mice infected with Hantaan virus. In the People's Republic of China, ribavirin significantly reduced mortality in patients with hemorrhagic fever with renal syndrome. Ribavirin has not been effective in animal models of filoviral and flaviviral infections. The only important adverse effect of ribavirin in humans is manageable, reversible anemia. PMID- 2546249 TI - Actions of complement on Junin virus. AB - Fresh sera from normal rhesus monkeys, guinea pigs, and rabbits inactivated 90% 99% of the infectivity of Vero cell-passaged, attenuated strains of Junin virus (JV) within 60 minutes. Selective depletion studies showed that inactivation occurred by the classical complement pathway. Complement had little effect on virulent JV strains. Adsorption of the fresh sera with JV-infected Vero cells showed that inactivation was not mediated by low levels of antibodies in normal sera. The cells used for propagation of the virus affected inactivation: virus passaged in Vero cells (a continuous African green monkey kidney line) was more susceptible than virus passaged in FRhL-2 cells (a diploid strain derived from fetal rhesus monkey lung). Complement was important for in vitro neutralization of virulent JV strains by immune sera but was unnecessary for neutralization of attenuated strains. Thus, complement may be important in host resistance to Argentine hemorrhagic fever in two ways: first, complement activation may contribute to the attenuated phenotype of some strains; and second, complement may be necessary for efficient neutralization of virulent strains. PMID- 2546250 TI - Prevention of Rift Valley fever in rhesus monkeys with interferon-alpha. AB - Prophylactic and therapeutic efficacy of recombinant leukocyte A interferon (rIFN alpha A) and Sendai virus-induced human leukocyte interferon (HuIFN-alpha) administered intramuscularly to Rift Valley fever virus (RVFV)-infected rhesus monkeys was studied. Clinical, virologic, immunologic, and hemostatic parameters were monitored. Five daily inoculations of 5 X 10(5) units of either interferon product per kilogram of body weight, initiated 24 hours before or 6 hours after RVFV infection, prevented or greatly suppressed viremia. No clinical signs of disease or laboratory evidence of impaired hemostasis was observed. Serum neutralizing antibody to RVFV was detected within 6 days of virus inoculation. Prophylactic administration of 5 X 10(4) or 5 X 10(3) units of rIFN-alpha A per kilogram also limited viremia, hepatocellular damage, and hemostatic derangement. Untreated, RVFV-infected, control monkeys developed high-titered viremia, clinical disease, and impaired hemostasis. These data suggest that rIFN-alpha A and HuIFN-alpha are effective in protecting RVFV-infected rhesus monkeys from viremia and hepatocellular damage and may be beneficial in human RVF infection. PMID- 2546251 TI - Immunocytochemical demonstration of cytochrome c oxidase as a marker for renal oncocytes and oncocytomas. AB - Using an antibody raised against the mitochondrial enzyme cytochrome c oxidase (EC 1.9.3.1), it was possible to visualize microoncocytomas, oncocytic tubules and even single oncocytes in the renal cortex of rats treated with N nitrosomorpholine, and to localize the lesions to the collecting duct of the nephron. In human specimens, the antibody also stained oncocytomas and oncocytic tubules in the surrounding macroscopically unaffected cortex. Thus, this antibody may be used for detection as well as for investigation of the development of oncocytic lesions in various species. PMID- 2546252 TI - Spontaneous nephroblastoma in a strain CE/J mouse. A case report. PMID- 2546253 TI - Eight year study of viral isolates from cot deaths in Glasgow. AB - Over an eight year period 21 different viral strains excluding polioviruses were isolated in 44 (19%) of 237 cot deaths. The percentage of viral positive cases was significantly greater in over 16 week age groups and in cultures obtained less than 24 hours after death. PMID- 2546254 TI - Further liaisons with sperm. PMID- 2546255 TI - Effect of platelet-activating factor on platelets and vascular endothelium. PMID- 2546256 TI - Endothelial cell interactions with heparin. PMID- 2546257 TI - [Current concepts on diabetic diets]. PMID- 2546258 TI - Edema formation in spinal nerve roots induced by experimental, graded compression. An experimental study on the pig cauda equina with special reference to differences in effects between rapid and slow onset of compression. AB - Edema formation in spinal nerve roots of the pig cauda equina was studied following experimental compression at various pressure levels, durations, and rates of onset, using a fluorescence microscopic technique. The time-pressure thresholds for the occurrence of edema in the nerve roots were: following rapid onset of compression (0.05-0.1 seconds), 2 minutes at both 50 mm Hg and 200 mm Hg, and following slow onset of compression (the pressure was slowly increased during 15-20 seconds), 2 hours at 50 mm Hg and 2 minutes at 200 mm Hg. Generally, the edema formation was more pronounced after rapid than after slow onset of compression. The data in this study also indicate that intraneural edema might be more easily formed in nerve roots than in peripheral nerves after compression injury. PMID- 2546259 TI - Evaluation of the risk to health in the exposure to asbestos, by means of scanning electron microscopy. PMID- 2546260 TI - Viral heart disease. PMID- 2546261 TI - The significance of autoantibodies against the ADP/ATP carrier for the pathogenesis of myocarditis and dilated cardiomyopathy--clinical and experimental data. PMID- 2546262 TI - Role of natural killer cells in experimental murine myocarditis. PMID- 2546263 TI - Role of cytotoxic T cells in experimental myocarditis. PMID- 2546264 TI - Anti-viral and anti-myocyte antibodies in experimental myocarditis. PMID- 2546265 TI - Heterotransplantation of human ductal pancreas carcinoma onto NMRI nu/nu mice. PMID- 2546266 TI - Characterization of human ovarian cancer lines with a panel of monoclonal antibodies (MoAbs). PMID- 2546267 TI - Dose response relationships in two xenografted human gliomas after fractionated radiotherapy. PMID- 2546268 TI - Fractionated irradiation under ambient conditions of human NSCLC transplanted into nude mice: effect of different fractionation schedules and of radiotherapy and vindesine. PMID- 2546269 TI - Critical valuation of the subrenal capsule (SRC) assay and its use in experimental therapy. PMID- 2546270 TI - Characterization and chemotherapy of human soft tissue sarcoma (STS) lines grown in nude mice. PMID- 2546271 TI - Treatment of high grade glioma xenografts with nitrosoureas in nude mice. PMID- 2546272 TI - Tumor necrosis factor--a new therapeutic approach. Preclinical studies in nude mice. PMID- 2546273 TI - Influence of interleukin II on xenotransplanted grade 3 to 4 glioma in nude mice. PMID- 2546274 TI - Physiology and pharmacology of alpha and beta receptors in the eye. PMID- 2546275 TI - Low molecular weight heparin treatment in a pregnant woman with allergy to standard heparins and heparinoid. PMID- 2546276 TI - Superoxide anion scavenging property of dipyridamole. PMID- 2546277 TI - The mode of action of low molecular weight heparin preparation (PK10169) and two of its major components on thrombin generation in plasma. AB - We studied the mode of action of the low molecular weight heparin PK10169 and two of its constituent fractions: EMT 966 High Molecular Weight Fraction and EMT 967 Low Molecular Weight Fraction. EMT 966 like standard heparin, acts primarily on thrombin formed and not on prothrombinase (S type heparin). In contrast EMT 967 has no direct effect on thrombin. At high concentrations, it inhibits the prothrombinase complex (P type heparin). PK10169, that contains the two EMTs shows both activities: antithrombin and antiprothrombinase (mixed type heparin). The addition of increasing amounts of EMT 967 to a constant amount of EMT 966 does not influence the breakdown constant of endogenous thrombin which is determined by the concentration of EMT 966 only. This demonstrates the absence of competition for AT III between the two components of PK10169. In platelet rich plasma, EMT 966 inhibits and postpones thrombin generation more efficiently than unfractionated heparin, probably because it is less sensitive to neutralization by platelet components (platelet factor 4). Amounts of EMT 967 that hardly inhibit thrombin generation in platelet rich plasma enhance the effect of EMT 966 probably by neutralizing platelet factor 4. PMID- 2546278 TI - R 68 070: thromboxane A2 synthetase inhibition and thromboxane A2/prostaglandin endoperoxide receptor blockade combined in one molecule--I. Biochemical profile in vitro. AB - R 68 070 or (E)-5-[[[(3-pyridinyl)[3-(trifluoromethyl)phenyl]- methylen]amino]oxy] pentanoic acid (Janssen Research Foundation, Belgium) combines specific thromboxane A2 (TXA2) synthetase inhibition with TXA2/prostaglandin endoperoxide receptor blockade in one molecule. In vitro, the compound specifically inhibits the production of TXB2 from [14C] arachidonic acid by washed human platelets (IC50 = 8.2 X 10(-9) M) and by platelet microsomes (IC50 = 3.6 X 10(-9) M), of MDA (IC50 = 1.91 X 10(-8) M) and of TXB2 (IC50 = 1.47 X 10(-8) M) by thrombin-coagulated human platelet-rich plasma (P.R.P.) and whole blood respectively and increases the levels of PGD2, PGE2, PGF2 alpha and 6-keto PGF1 alpha. The activity of cyclo-oxygenase-, prostacyclin synthetase-, 5-, 12- and 15-lipoxygenase-enzymes are not affected. Additionally, R 68 070 inhibits human platelet aggregation in P.R.P. induced by U 46619 3 X 10(-7) M to 2 X 10( 6) M (IC50 = 2.08 X 10(-6) M to 2.66 X 10(-5) M, collagen 0.5 to 2 micrograms/ml (IC50 = 2.85 X 10(-6) M to 4.81 X 10(-5) M), arachidonic acid 7.5 X 10(-4) M to 2 X 10(-3) M (IC50 = 2.1 X 10(-8) M to 3.3 X 10(-8) M) and the U 46619 (1 X 10(-7) M)-induced accumulation of [32P] phosphatidic acid (IC50 = 5.24 X 10(-7) M) in washed human platelets.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546279 TI - Low molecular weight heparin Novo (LHN-1) does not cross the placenta during the second trimester of pregnancy. AB - Unfractionated heparin (UF-H) has been the drug of choice for the treatment of thromboembolic disorders during pregnancy. Low molecular weight heparin (LMW-H) preparations may present some advantages over UF-H. They have longer half-lives and a better bioavailability after subcutaneous (s. c.) injection and may cause less bleeding. It has not yet been established whether LMW-H Novo (LHN-1) crosses the placenta. 17 women admitted for abortion during the second trimester of pregnancy (induced by application of prostaglandine PGE2 gel at a concentration of 0.25 mg/ml into the cervix) were given s. c. 35 anti-Xa units per kg of body weight of LHN-1 (Novo). 10 patients not receiving LHN-1 and their fetuses served as a control group. 7 women in whom the time interval between injection of LHN-1 and expulsion of the fetus was less than 3 h or more than 7 h were excluded from further study. In one fetus blood collection failed. Anti-Xa and anti-IIa levels increased approximately ten-fold in women receiving LHN-1 [anti-Xa units/ml from 0.02 +/- 0.01 (mean +/- SD) to 0.17 +/- 0.01, p less than 0.001; anti-Ha units/ml from less than 0.01 +/- 0.01 to 0.07 +/- 0.03], but remained below the detection limit in their fetuses as well as in the women and fetuses of the control group. We conclude that LHN-1 at these doses does not cross the placenta during the second trimester of pregnancy to suggest that LHN-1 may be a safe alternative to heparin in the management of the thromboembolic complications during pregnancy. PMID- 2546280 TI - Functional assays for protein C activity and protein C inhibitor activity in plasma. AB - An assay system for protein C (PC) activity and PC-inhibitor in plasma was developed. The assay was based on: (1) binding of PC to wells of a microtiter plate coated with a murine monoclonal anti-PC antibody (C3) that did not interfere with the activity or activation of PC; (2) activation of immobilized PC with Protac C; (3) incubation with or without a source of activated PC inhibitor; and (4) measurement of amidolytic activity using the substrate S-2366. The activity assay was specific for PC and sensitive to less than 1 microliter of plasma or 4 ng PC. Inhibition of activated PC by plasma followed pseudo first order kinetics. Heparin caused a dose dependent increase in the inhibition rate with half maximal stimulation at approximately 3 U/ml and maximal stimulation at heparin concentrations greater than or equal to 10 U/ml. This assay is suitable not only for determination of functional plasma levels of PC and PC inhibitor activities but also for kinetic studies of inhibition of activated PC in complex systems, such as plasma. Studies showed that urokinase interfered with the inhibition of APC by plasma inhibitor(s). PMID- 2546281 TI - Identification of mRNA coding for factor VII protein in human alveolar macrophages--coagulant expression may be limited due to deficient postribosomal processing. AB - Clotting factors synthesized by monocytes and macrophages may initiate coagulation reactions during inflammation. Functional vitamin K-dependent coagulation factors have been found to be associated with human monocytes/macrophages, but there are no reports identifying mRNA coding for vitamin K-dependent proteins in these cells. In the present studies, factor VII mRNA was found in total RNA extracted from freshly isolated human alveolar macrophages using hybridization with a complementary DNA probe. On the other hand, vitamin K-dependent carboxylase activity which is required for postribosomal modification of the protein, was not detectable in the macrophages before or after culture, and human blood mononuclear leukocytes also lacked this enzyme activity. Control human and rat hepatoma cells exhibited high levels of carboxylase activity within the same experiments. Using sensitive kinetic assays, no increase in factor VII activity was detected during culture of alveolar macrophages under conditions promoting 1.78 +/- .24 (n = 8) fold increases of tissue factor activity. These findings with freshly isolated cells demonstrate that alveolar macrophages synthesize factor VII mRNA in vivo. However, the mRNA was found in the absence of evidence for gamma-carboxylase activity or processing of the factor into a functional clotting enzyme. The results imply that functional expression of any synthesized coagulation factor VII in alveolar macrophages may be limited or prevented due to a cellular deficiency at the level of postribosomal processing. PMID- 2546282 TI - The relative antithrombotic effectiveness of heparin, a low molecular weight heparin, and a pentasaccharide fragment in an animal model. AB - The antithrombotic efficacy of unfractionated heparin (UFH), a low molecular weight heparin (LMWH) and a synthetic pentasaccharide (PENTA) has been compared in an animal model for stasis thrombosis. We have also compared the relative ability of these three agents to impair thrombin generation in vitro and in vivo, and measured their effects on anti-Xa activity and thrombin clotting times. UFH, LMWH and PENTA all had the capacity to impair thrombogenesis, although there were marked differences in their relative effectiveness. Reduction of thrombin generation to 20% of control values was closely correlated with the prevention of thrombosis after 20 minutes' stasis, but this was only achieved with UFH. The same dry weight dose of LMWH or PENTA reduced thrombin generation to about half control values, and neither significantly impaired thrombus formation after 20 minutes' stasis. Impaired thrombin generation correlated better than anti-Xa activity with prevention of stasis thrombosis. In this model, UFH was clearly superior to LMWH and PENTA as an antithrombotic agent. PMID- 2546283 TI - Immunogold electron-microscopic localization of calpain I in human erythrocytes. AB - With a view to understanding the function of calpain I (EC 3.4.22.17) in vivo, the localization of the enzyme was studied by immunoelectron microscopy in human erythrocytes. Thin sections of the cells embedded in Spurr's resin were exposed to solutions of monoclonal anti-calpain I or control antibodies, biotin antimouse IgG antibodies, and streptavidin-gold. Most of the calpain I (93%) was found to be distributed throughout the cytoplasm, and only 7% of the gold particles were associated with the erythrocyte membrane. Erythrocytes were Ca2+-loaded by means of the calcium ionophore A23187, and the rise in intracellular [Ca2+] was demonstrated both by crenation of the cells, and by activation of calpain which was detected by immunoblotting. The proportions of cytosolic and membrane-bound gold labelling were, however, not altered by Ca2+-loading. These results are not consistent with the hypothesis that activation of calpain requires membrane binding. PMID- 2546284 TI - In vitro effects of ethanol on rabbit platelet aggregation, secretion of granule contents, and cyclic AMP levels in the presence of prostacyclin. AB - Ethanol, at physiologically tolerable concentrations, inhibits platelet responses to low concentrations of collagen or thrombin, but does not inhibit responses of washed rabbit platelets stimulated with high concentrations of ADP, collagen, or thrombin. However, when platelet responses to high concentrations of collagen or thrombin had been partially inhibited by prostacyclin (PGI2), ethanol had additional inhibitory effects on aggregation and secretion. These effects were also observed with aspirin-treated platelets stimulated with thrombin. Ethanol had no further inhibitory effect on aggregation of platelets stimulated with ADP, or the combination of ADP and epinephrine. Thus, the inhibitory effects of ethanol on platelet responses in the presence of PGI2 were very similar to its inhibitory effects in the absence of PGI2, when platelets were stimulated with lower concentrations of collagen or thrombin. Ethanol did not appear to exert its inhibitory effects by increasing cyclic AMP above basal levels and the additional inhibitory effects of ethanol in the presence of PGI2 did not appear to be brought about by further increases in platelet cyclic AMP levels. PMID- 2546285 TI - Monoclonal antibodies to tissue-type plasminogen activator which prolong its clearance in vivo. AB - Three murine monoclonal antibodies to tissue-type plasminogen activator (t-PA) were evaluated for their effects on the binding of iodinated t-PA to cultured human hepatoma cells (Hep G2), and on extending the half-life of t-PA injected into rabbits. Two of the antibodies, AE5 and EG2, significantly inhibited t-PA binding in vitro, and extended the in vivo half-life of t-PA four to five-fold. A third antibody, BA10, which had a much smaller inhibitory effect on t-PA binding, had no influence in extending t-PA's half-life. MOPC-21, a control antibody not directed to t-PA, had no effect on either test. Our results are the first to correlate different compounds' effects on t-PA binding with their ability to retard t-PA clearance in vivo, and provide additional evidence for the importance of a liver cell receptor in the t-PA clearance process. PMID- 2546287 TI - [Phyllodes tumor]. AB - This very rare breast tumour has been diagnosed in five cases, four females and one male, at our hospital during the last two years. The females all presented tumours with a diameter ranging from 5 to 15 cm. Age at diagnosis ranged from 21 to 54 years. We focus on diagnostic challenges, since 3 of our patients were recently treated by removal of a breast tumour diagnosed morphologically as fibroadenomatosis. Our patients underwent different surgical therapy, and we focus on local treatment modalities advocated in recent literature. Systemic chemotherapy, endocrine treatment and/or radiation therapy, in the adjuvant setting or in advanced disease, do not increase survival in patients with malignant phyllodes tumour. PMID- 2546288 TI - [Cytomegaloviruses. Prevalence of cytomegalovirus infections in married couples]. PMID- 2546289 TI - [Lisinopril (Zestril)--a new ACE inhibitor]. PMID- 2546286 TI - DN 9693: a phosphodiesterase inhibitor with a platelet membrane effect. AB - We have examined the in vitro effects of DN 9693 (piperidinylimidazo quinazolinone) on various aspects of platelet reactivity. Our results are consistent with its known function as a phosphodiesterase inhibitor in that it increased platelet cyclic AMP, particularly in conjunction with an adenylate cyclase stimulator, and exerted a profound inhibitory effect on platelet aggregation responses to a variety of agonists. DN 9693 also inhibited ristocetin induced platelet agglutination (RIPA). We therefore examined its effect on ristocetin co-factor assays and on the binding of a monoclonal antibody (McAb) to platelet membrane glycoprotein Ib (GPIb). The drug inhibited the binding of the monoclonal antibody in a dose-dependent manner. This suggests an effect of the drug on the platelet surface membrane with reduced expression of GPIb. Our results indicate that in addition to its anticipated inhibitory effect on platelet aggregation, DN 9693 may also inhibit platelet adhesion. PMID- 2546290 TI - Canine mammary tumour disease: role of hormones in pathogenesis and treatment. PMID- 2546291 TI - MK-801 prevents cognitive and behavioral deficits produced by NMDA receptor overstimulation in the rat hippocampus. AB - The overstimulation of receptors for L-glutamate, particularly those of the N methyl-D-aspartate (NMDA) type, has been suggested to play a role in mediating damage in a variety of neurodegenerative conditions or disorders ranging from ischemia/hypoxia to senile dementia of the Alzheimer's type (SDAT). We report here that the functional deficits and histological damage mediated by the overactivation of NMDA receptors in the Fischer 344 rat hippocampus can be blocked effectively by systemic administration of the noncompetitive NMDA antagonist, MK-801. These results suggest that MK-801 may be effective clinically in attenuating memory loss and hippocampal damage in disorders associated with the overstimulation of NMDA receptors. PMID- 2546292 TI - Lung cell reactions in guinea pigs after inhalation of asbestos (amosite). AB - To study the long-term effect of inhaled asbestos, guinea-pigs were exposed to airborne amosite at a concentration of 49 mg/m3, 2 h/day for 3 or 6 weeks and examined up to 2 years after exposure. Macrophages in lung lavage fluid (LLF) were increased at 16, 24 and 93 weeks and lymphocytes at 24 weeks. Examination of lung wall cells (LWC) 2 years after exposure compared to cells obtained by LLF showed higher proportions of LWC lymphocytes and neutrophils. Percoll gradient centrifugation of these cells showed a higher proportion of high density macrophages in LLF from the amosite-exposed animal and an increased number of low density lymphocytes in the LW. Cathepsin D was increased in LLF at 8 and 24 weeks and in alveolar macrophages 24 weeks and 2 years after exposure. Fibroblast cultures exposed to LLF did not show any statistical significant changes in their collagen synthesis. Histology 93 weeks after exposure showed macrophage and mediastinal lymph node accumulation of asbestos, as well as collagen in alveolar walls. Granulomas were found in the vicinity of blood vessels and in connection with the bronchioles. The data suggest that amosite at low doses ultimately causes fibrosis with a reaction pattern different from that seen in silicosis. The inflammation and fibrosis seems to develop only within the interstitium. PMID- 2546293 TI - Metabolism of N-nitroso-hydroxyethyl-alkylamine phosphate esters in the rat. AB - The metabolism of 1-(N-methyl-N-nitrosamino)-ethylphosphate and 1-(N-ethyl-N nitrosamino)-ethylphosphate in the rat was investigated. The determination of blood clearance, organ clearance, excretion of parent compounds in the urine and the exhalation of radiolabeled CO2 originating from a nitrosaminophosphate demonstrated a rapid metabolism of the compounds. The high activity of alkaline phosphatase in kidney caused a very rapid degradation of the nitrosamino phosphates in kidney homogenate, whereas the compounds were relatively stable in liver homogenate and serum. We, therefore, suggest a rapid degradation of such nitrosamino conjugates, if they are formed at all, in vivo. PMID- 2546294 TI - Lack of effect of a neurotoxin from the scorpion Buthus martensi Karsch on nerve fibers of this scorpion. AB - A neurotoxin (BmK I) was purified from the venom of the scorpion species Buthus martensi Karsch. Effects of this toxin on the excitability of the abdominal nerve fibers of the same scorpion were examined. The toxin had no effect at all on the action and resting potentials recorded intracellularly even at a concentration as high as 100 microM. A similar result was obtained through optical measurements of the action potential using a potential sensitive dye. Sea anemone toxin II (8 microM) had no effect on nerve excitability either. However, tetrodotoxin (50 nM) reversibly suppressed the action potential and grayanotoxin II (20 microM) induced a sustained depolarization of the nerve membrane which resulted in a reversible suppression of the action potential. BmK I at a concentration of 0.1 microM greatly prolonged the action potential in the crayfish giant axon. We conclude that the Na channel of nerve fibers of this scorpion is totally insensitive to the neurotoxin in this scorpion's venom. PMID- 2546295 TI - Respiratory effects of brevetoxin and saxitoxin in awake guinea pigs. AB - Ptychodiscus brevis toxin (brevetoxin) is associated with 'Florida red tide' and cause neurotoxic shellfish poisoning. Saxitoxin is the agent of paralytic shellfish poisoning. Clinical reports of human intoxication suggest that both toxins affect the respiratory system. The toxins were administered by slow intravenous infusion. The effects of the toxins on respiratory function of awake guinea pigs in a pressure plethysmograph were studied. Both toxins caused lactic acidosis of unknown etiology, which was compensated for by increased minute volume with brevetoxin (PbTx-3)- but not with saxitoxin-intoxicated animals. In general, brevetoxin increased ventilation, before respiratory failure, while saxitoxin had a depressive effect on ventilation. Airways resistance was not increased, nor was dynamic compliance decreased during intoxication, although the data suggest that respiratory system failure was the primary cause of death. The responses seen in these experiments are consistent with the dissimilar molecular actions of these toxins. PMID- 2546296 TI - In-vitro spin-trapping of free radicals produced during trichloroethylene and diethylether metabolism. AB - Free-radical production during the metabolism of various xenobiotics represents a frequent mechanistic explanation for their toxicity. We tested the hypothesis of production of free radicals from two solvents, diethylether and trichloroethylene (TRI), and from two metabolites of TRI, namely trichloroethanol (TCE) and trichloroacetic acid (TCA). The formation of free radicals was detected by electron spin resonance spectroscopy (ESR), using a spin-trapping agent, alpha-(4 pyridyl-1-oxide)-N-tert-butyl-nitrone (POBN). Two experimental models were used. The first was a chemical model using Fenton's reagent, a mixture of Fe(II) chelator and H2O2, for which the normal reaction is OH. production, and the second, a preparation from rat liver and brain microsomes containing NADPH and achieving enzymatic oxidation of the solvents. After addition of diethylether, free-radical production was demonstrated under the two experimental conditions. This free radical probably derived from the parent molecule by hydrogen abstraction. TRI and TCE additions to the Fenton system suppressed normal OH. production whereas this production was increased after TCA addition. The addition of TCE to the microsomal preparations was followed by free-radical production which could derive either from the parent molecule or from other sources, e.g. from membrane degradation, with a preference for the first hypothesis because of the characteristics of the signal. This result was not observed after addition of TRI or TCA. In conclusion, these preliminary results confirm the validity of the hypothesis of production of free radicals from diethylether, but they are less consistent for TRI as this production was observed only after addition of TCE; this result is interesting, however, as TCE is considered to play a major role in the toxicity observed after TRI exposure in humans. PMID- 2546297 TI - The effects of 1,3-dinitrobenzene and mono-(2-ethylhexyl)phthalate on hormonally stimulated lactate and pyruvate production by rat Sertoli cell cultures. AB - Lactate and pyruvate production by rat Sertoli cell cultures was measured after treatment with two toxicants (1,3-dinitrobenzene, DNB; mono-(2 ethylhexyl)phthalate, MEHP) in the presence or absence of follicle-stimulating hormone (FSH) or dibutyryl-cAMP (dbcAMP). 1,3-DNB together with FSH or dbcAMP produced increases in media lactate and pyruvate concentrations significantly greater than those induced by individual treatments. MEHP in conjunction with either hormone also produced greater increases in lactate concentrations compared with separate additions. However, MEHP produced a significant diminution in the hormonal stimulation of pyruvate secretion. Thus, 1,3-DNB appears to act independently of these hormones in stimulating the secretion of both metabolic products, whereas MEHP interferes with the hormonal stimulation of pyruvate (but not lactate) production. PMID- 2546299 TI - The influence of T cell depletion on recovery of T cell proliferation to herpesviruses and Candida after allogeneic bone marrow transplantation. AB - To test the influence of T cell depletion of the marrow in allogeneic bone marrow transplantation on functional T cell recovery, in vitro lymphocyte proliferation tests (LPTs) to microbial antigens were regularly performed in 23 recipients of normal BM and in 25 patients receiving BM with a fixed low number of T cells (1 x 10(5) T cells/kg body weight; recipients of T-depleted BM). The long-term recovery of positive LPT to at least 1 of the 4 tested microbial antigens- Candida, herpes simplex virus (HSV), varicella-zoster virus (VZV), and cytomegalovirus--was nearly similar in both groups: 16/23 versus 18/25. Recovery of LPT to Candida and HSV in the first 3 months appeared to be greatly influenced by prophylactic measures; only 2/23 recipients of normal BM, receiving amphotericin B, showed a positive LPT to Candida versus 13/25 recipients of T depleted BM (P less than 0.01). In contrast, only 1/23 seropositive recipients of T-depleted BM, receiving acyclovir, showed a positive LPT to HSV versus 9/22 recipients of normal BM (P less than 0.05). A positive LPT to CMV in the first 3 months was found in 9/9 seropositive recipients of normal BM, versus in 5/11 seropositive recipients of T-depleted BM (P less than 0.05). Five of the 6 patients with a negative LPT died of CMV-interstitial pneumonia versus 1/14 with positive LPT (P less than 0.01). We conclude that in CMV-seropositive recipients of allogeneic BM, T cell depletion of the graft affects the early recovery of T cell proliferation to CMV, which is associated with a higher risk of fatal CMV interstitial pneumonia. PMID- 2546298 TI - Response to platelet-activating factor in human platelets stored and aged in plasma. Decrease in aggregation, phosphoinositide turnover, and receptor affinity. AB - Human platelet concentrates were stored in polyolefin bags at 22 to 24 degrees C on a horizontal shaker for up to 8 days. At different intervals, aliquots of platelet-rich plasma (PRP) were removed aseptically and five variables, i.e., platelet counts, morphology, platelet-activating factor (PAF)-stimulated aggregation, phosphoinositide turnover, and [3H]PAF binding to platelet receptors, were studied. The number of platelets did not change during the 8 days of storage. Scanning electron microscopy of the platelets revealed a gradual morphologic change from biconcave flat discs to irregular, crenated forms. The PAF-induced aggregation of platelets declined with time of storage. A decrease to 50 percent of the Day 1 aggregatory response to PAF was evident on Day 2, and there was a further decline to about 20 percent by Day 6. Similarly, PAF receptor coupled phosphoinositide turnover, as monitored by 32P incorporation into individual phosphoinositides, decreased dramatically with storage. After 2 to 3 days of storage, the phosphoinositide turnover was reduced to 50 percent of the original response, and it continued to decline to about 25 percent of original response by Day 5 or 6. The binding of [3H]PAF to washed human platelets indicated subtle changes between Days 2 and 4, which became more noticeable by Day 6. These results have raised the possibility of changes in the number of the receptors and/or their affinity for the ligand during storage. We conclude that although the number of platelets was maintained during storage for 8 days, a general deterioration of their responses to PAF occurred at the levels of cell surface receptor, transmembrane signaling (phosphoinositide turnover), and response (aggregation). PMID- 2546300 TI - Transmission of Epstein-Barr virus by a transplanted kidney, with activation by OKT3 antibody. PMID- 2546302 TI - Multiple bradykinin B2 receptors. PMID- 2546301 TI - Rapid detection of human cytomegalovirus DNA in peripheral blood leukocytes of viremic transplant recipients by the polymerase chain reaction. AB - Peripheral blood leukocyte samples (n = 458) of 24 bone marrow transplant and 52 kidney transplant patients were examined weekly for the presence of human cytomegalovirus (HCMV) using an improved culture technique (DEAFF; detection of early antigen fluorescent foci). In total 5 (21%) bone marrow transplant and 11 (21%) kidney transplant patients developed a viremia. Patients' samples were investigated for the presence of HCMV DNA using an in vitro DNA amplification technique, the polymerase chain reaction (PCR). From the statistically evaluable viremic patients (n = 13), 110 blood samples were analyzed. In 5 of these patients, the DEAFF and PCR led to identical results. In 8 patients however the PCR was more sensitive, i.e. HCMV DNA was detected for a longer period of time. Applying statistical analysis using the McNemar test, this result was significant (P less than 0.05). The PCR applied on leukocyte samples did not detect HCMV DNA in viruric patients without viremia. Moreover, the current PCR never led to positive results with peripheral blood leukocyte samples of healthy seropositive or seronegative controls. Since the PCR can be performed in 6 hr, this technique will contribute to rapid detection of HCMV DNA in peripheral blood leukocytes and therefore to optimal clinical management of HCMV-infected transplant recipients. PMID- 2546303 TI - Molecular recognition of inositol polyphosphates by intracellular receptors and metabolic enzymes. AB - The discovery that inositol lipids are fundamentally involved in cell signalling has been one of the most significant recent advances in cell biology. In particular, there is now evidence that certain products of polyphosphoinositide metabolism play second messenger roles in most cells. Inositol 1,4,5 trisphosphate and perhaps inositol 1,3,4,5-tetrakisphosphate bind to specific receptors and regulate Ca2+ release from, and movement between, intracellular stores. The synthesis of novel analogues of these second messengers is now providing clues to the structural requirements at such receptors as well as for molecules with stability towards metabolic enzymes. Stefan Nahorski and Barry Potter discuss these developments with a view to future pharmacological intervention at these sites. PMID- 2546304 TI - The histamine H3 receptor: a general presynaptic histaminergic regulatory system? AB - The histamine H3 receptors were initially identified as presynaptic autoreceptors in the brain. However, recent research described here by Jan van der Werf and Hendrik Timmerman demonstrates that H3 receptors are associated with multiple functions, and their location is not confined to the CNS. PMID- 2546305 TI - [A receptor of dihydropyridine blockaders of Ca2+ influx in human embryonic fibroblasts]. AB - The plasma membrane of human embryonic fibroblasts was shown to contain receptor binding sites for 1,4-dihydropyridine (DHP) Ca2+ entry blockers. In a subconfluent culture grown in serum medium the content of the DHP receptor amounted to 1.2 +/- 0.3 pmol per 10(6) cells. With progression to confluency this value decreased up to 0.28 +/- 0.08 pmol per 10(6) cells. The DHP binding capacity was shown to be affected by the presence of growth factors in culture medium. In a subconfluent culture of serum-deprived cells the content of DHP binding sites increased 1.9 fold, the steady-state level being achieved within 3 days in culture. The serum gradually reversed this process, and the DHP receptor density approached the initial level within 3 days. PMID- 2546306 TI - Immune response of chicks vaccinated with fowl pox virus vaccines by the feather follicle method. PMID- 2546307 TI - [Structural organization of blood plasma lipoproteins and their interaction with the cell]. AB - Up-to-date information on the structural organization of basic blood plasma lipoproteins and their interaction with a cell is presented. Different types of lipoprotein modifications which make changes in lipoprotein interactions with the receptors of macrophages, fibroblasts and other cells are shown. The participation of different types of receptors in the process of interaction with native and modified lipoproteins is emphasized. PMID- 2546308 TI - [Activation of succinate oxidase in the inner membrane of rat liver mitochondria]. AB - It is shown that the process of activation of succinate oxidase from inner membranes of the rat liver mitochondria by succinate and malonate is specific for the succinate dehydrogenase component of oxidase. These activation constants are comparable with those found by other authors in activation of succinate dehydrogenase and succinate oxidase from oxaloacetate-preincubated submitochondrial fragments of the bull heart. Probably, the 4-fold activation of succinate oxidase from inner membranes of the liver mitochondria reported in this paper depends on separation of endogenous oxaloacetate from the succinate dehydrogenase component of oxidase. PMID- 2546309 TI - [Morphological differentiation and change in the lipid composition of neuroblastoma C1300 cells treated with gangliosides]. AB - The effect of exogenous gangliosides on the morphological differentiation of neuroblastoma 1300 N18 cells was studied Simultaneously the content of gangliosides and lipid composition of the cells was investigated. Gangliosides were shown to increase the quantity of cells with long neurites. This effect depended on the dose of gangliosides. The addition of 50 and 100 micrograms of gangliosides per 4 ml of serum-free culture medium increased the quantity of cells with neurites by 38 and 63.4%, respectively. The level of morphological differentiation in cells cultivated with gangliosides was higher than in cells incubated with 5'-bromodeoxyuridine. Noticeable quantities of lysophosphatidylcholine (absent in the control) appeared in ganglioside-treated cells and the level of cholesterol increased. The amount of other lipid compounds in cells differentiated in the presence of gangliosides was similar, but not identical to the quantity of lipid compounds in cells differentiated by 5' bromodeoxyuridine and by the serum-free medium. PMID- 2546310 TI - Germ cell tumors of the testicle or 'orchidomata'. AB - An unusual case of a double testicular tumor, with different histology and the same marker chromosome, led to a search of the literature for cases of testicular tumors that were double, bilateral, or familial. The literature on abnormal chromosomes in tumors of the testis is also reviewed. After a discussion of the facts, it is suggested that most of the histological variants of germ cell tumors of the testicle are so closely related that they could be grouped together under the title of 'orchidomata'. PMID- 2546311 TI - Adrenal myelolipoma simulating virilizing adrenal tumor. AB - We describe a case of adrenal myelolipoma that simulated clinically and biochemically a virilizing adrenal tumor. The tumor was removed surgically. The preoperative diagnosis of adrenal myelolipoma can be made by the characteristic appearance on ultrasonography, computerized tomography and in equivocal cases by guided needle biopsy. PMID- 2546312 TI - The influence of hydroxyapatite and pyrophosphate on the formation product of calcium oxalate at different pHs. AB - The nucleating effect of hydroxyapatite (HAP) and the inhibitory effect of pyrophosphate (PPi) on calcium oxalate crystallization have been studied at different pH's in solution metastabely supersaturated with respect to calcium oxalate but saturated with respect to HAP. Crystallization was monitored by a decrease of calcium in the supernatant and formation products were calculated. At a pH above 6.0 already minimal HAP concentrations proved to be a suitable substrate for heterogeneous nucleation and growth of calcium oxalate. PPi showed a pronounced inhibitory effect on spontaneous as well as on HAP induced crystallization of calcium oxalate, this effect being highly pH dependent. HAP was found to neutralize the inhibitory effect of PPi in a molar ratio of 10:1. PMID- 2546313 TI - Urolithiasis inhibitors and calculus nucleation. AB - The possible inhibitors of heterogeneous nucleation were investigated. The effects of magnesium, pyrophosphate, citrate and Chondroitin Sulphate on calcium phosphate or uric acid heterogeneous nucleation of calcium oxalate were studied. It was found that whereas magnesium, pyrophosphate and citrate acted as effective inhibitors in the presence of calcium phosphate as heterogeneous nucleant, only chondroitin sulphate manifested important inhibitory effects when uric acid was the heterogeneous nucleant. PMID- 2546315 TI - The relation between orthophosphate and pyrophosphate in normal subjects and in patients with urolithiasis. AB - In calcium lithiasis, inhibitors have a significant effect in reducing the crystallization process. This work evaluated orthophosphate in a group of patients with calcium oxalate lithiasis, and in a control group. The study of orthophosphate and pyrophosphate, showed differences between stone formers and the control group. These results could be attributed to a failure in the renal transformation of orthophosphate into pyrophosphate. PMID- 2546314 TI - The effects of citrate on hydroxyapatite induced calcium oxalate crystallization and on the formation of calcium phosphate crystals. AB - The addition of different amounts of hydroxyapatite crystals (HAP) to a solution, metastably supersaturated with respect to calcium oxalate (CaOx) resulted in heterogenous crystallization at seed concentrations exceeding 0.2 mmol/l. The induction period varied between 1 and more than 8 h with the shortest period for a seed concentration of 2 mmol/l. Addition to the system of 1 and 2% of whole urine and citrate in concentrations corresponding to approximately 1% of that found in normal urine inhibited the crystallization for as long as 4 h. In a system supersaturated with respect to calcium phosphate (CaP) the total number of crystals was markedly reduced by citrate concentrations exceeding 0.5 mmol/l. The fractions of medium sized and large crystals were sharply reduced and small crystals predominated at higher citrate concentrations. This might indicate effects of citrate on both crystal growth and crystal aggregation. We conclude that increased citrate concentrations during treatment with alkali leads to a significant inhibition of CaOx growth on HAP as well as to a prevention of the formation of large CaP crystals from solutions supersaturated with respect to CaP. PMID- 2546317 TI - Equine herpesvirus. PMID- 2546316 TI - [Regulation of reparative processes in glaucoma surgery by collalysin]. AB - Collalysin, a proteolytic enzyme, selectively acting on the young connective tissue, is suggested to be used as part of the complex of measures to prevent excessive cicatrization after antiglaucoma surgery. Rabbit experiments with 3H proline confirm the efficacy of the new drug; a more manifest stable hypotensive effect of antiglaucoma surgery in patients with open-angle glaucoma recommend collalysin for clinical application. PMID- 2546318 TI - Possible impairment of rotavirus immune response in cattle infected with BLV. PMID- 2546319 TI - Antibody isotype responses in the serum and respiratory tract to primary and secondary infections with equine influenza virus (H3N8). AB - Serum antibody (IgGab, IgM and IgA) responses to primary and secondary infection with influenza A/equine/Newmarket/79 (H3N8) by nebulised aerosol were compared with local (nasopharyngeal and tracheal) antibody responses in ponies. Circulating IgGab antibody was of long duration after primary infection, whereas IgM responses were short-lived after both primary and secondary infections. The antigenic stimulation of secondary infection with equine influenza was sufficient to induce elevations of serum IgM and IgA in the presence of high levels of circulating IgGab. These results support the potential of virus-specific IgM measurement for the detection of recent exposure to virus in horses which have high levels of circulating IgGab. Unlike serum IgGab, nasal and tracheal wash antibody of this isotype did not show long duration after primary infection, but local antibody memory was demonstrated by anamnestic responses on rechallenge. Nasopharyngeal IgA developed later than IgGab and IgM, and was more durable. PMID- 2546320 TI - Production and characterization of monoclonal antibodies to bovid herpesvirus-4. AB - Thirty-five hybridoma cell lines secreting monoclonal antibodies (Mabs) against bovid herpesvirus-4 (BHV-4) strain V. Test were produced. These hybrid cells resulted from the fusion of SP2/0 myeloma cells with splenocytes of BALB/c mice previously immunized with purified BHV-4. A modified indirect fluorescent antibody test (IFAT) was applied as a screening procedure and was compared with an indirect enzyme-linked immunosorbent assay. The selected Mabs were tested by the same IFAT against a panel of BHV-4 field isolates and against bovine herpesvirus-1, bovine herpesvirus-2 and alcelaphine herpesvirus-1 (AHV-1). Comparison of BHV-4 field isolates with Mabs confirmed their close antigenic relationships, but slight antigenic differences were observed between different isolates. One of the Mabs also reacted against AHV-1, indicating an antigenic relationship between BHV-4 and AHV-1. None of the Mabs reacting with BHV-4 possessed neutralizing activity against the strain used for immunization. PMID- 2546321 TI - [X-ray morphologic characteristics of microcalcination in benign processes and cancer of the breast]. AB - Mammographic and morphologic examination was carried out in 136 females bearing microcalcinates in breast tissue. Morphologic examination identified benign tumors (mostly fibrous cysts) in 72.1% and cancer (mostly ductal and lobular carcinoma in situ or initial signs of invasion) in 27.9% of cases. Calcinates occurred mainly in the epithelium and incipient cancer complexes. The analysis of the data showed shape, structure and distinctness of contours of calcinates to be instrumental in differentiating between malignant and benign lesions. PMID- 2546322 TI - [Granulocyte alkaline phosphatase and myeloperoxidase in patients with ischemic heart disease]. AB - Alkaline phosphatase and myeloperoxidase was examined in patients with chronic ischemic heart disease and myocardial infarction. It was established that patients with myocardial infarction showed high values of alkaline phosphatase while the values of myeloperoxidase were low. Dynamic evaluation of alkaline phosphatase and myeloperoxidase of leucocytes allows to assess the course of myocardial infarction and efficacy of its treatment. PMID- 2546323 TI - [Myeloperoxidase deficiency of the peripheral blood neutrophils and the incidence of infectious complications in patients with acute myeloblastic leukemia]. AB - The myeloperoxidase activity of polymorphonuclear peripheral blood neutrophils was studied in 60 patients with acute myeloblastic leucosis. It was established that patients with low peroxidase activity, frequency of infectious complications was 3.7 times higher than in patients with normal peroxidase activity. Hence, reduced myeloperoxidase activity of polymorphonuclear neutrophils in acute myeloblastic leucosis causes a decrease of the bactericidal activity of the cells. PMID- 2546324 TI - [The use of deoxypeganin hydrochloride in the combined treatment of patients with diseases of the peripheral nervous system]. AB - A study is presented of the efficacy of 1% deoxypeganin hydrochloride solution in muscle lesions due to involvement of the peripheral nervous system in 19 patients in whom the drug was administered into the motor points of the involved muscles. In 13 similar patients the drug was given intramuscularly. Clinical and electromyographic studies indicate that local use proved more effective than traditional. PMID- 2546325 TI - [The convulsive syndrome in pancreatic insuloma]. PMID- 2546327 TI - N-hydroxy-N-arylacetamides. V. Differences in the mechanism of haemoglobin oxidation in vitro by N-hydroxy-4-chloroacetanilide and N-hydroxy-4 chloroaniline. AB - 1. Autoxidation of N-hydroxy-4-chloroaniline(I) in buffer pH 7.4 was rapid and yielded 4,4'-azoxybischlorobenzene, 4-chloronitrosobenzene, 4-chloronitrobenzene, and 4-chlorophenyl nitroxide. In contrast, autoxidation of N-hydroxy-4 chloroacetanilide(II) was very slow, since in ether and water 78 and 92%, respectively, had decomposed in six months. 2. Haemoglobin(HbO2)-catalysed autoxidation of (I) occurred at a molar ratio of haemoglobin-Fe2+ to (I) of less than 0.25 and was accompanied by ferrihaemoglobin(HbFe3+)-formation and oxygen consumption. Coupled oxidation of HbO2 with (I) occurred at a molar ratio of greater than 0.2 and was accompanied by liberation of oxygen and the formation of HbFe3+, haemoglobin-4-chloronitrosobenzene complex, HbO2, desoxyhaemoglobin, 4 chloronitrosobenzene, 4-chloronitrobenzene, 4-chloroaniline, 4,4' azoxybischlorobenzene, and 4-chlorophenyl nitroxide. At an equimolar ratio of 10( 3) M haemoglobin-Fe2+ to (I), 96% HbO2 was converted into HbFe3+ (50%) and haemoglobin-4-chloronitrosobenzene complex in the initial fast phase of the reaction, but only 34% of the bound oxygen was liberated, the rest was sequentially reduced to water. (I) completely disappeared, and 4 chloronitrosobenzene was the major metabolite, mainly bound to haemoglobin. 3. Chemical oxidation of (II) by PbO2 in benzene produced acetyl 4-chlorophenyl nitroxide, whose spontaneous decomposition gave 38% 4-chloronitrosobenzene, 33% N acetoxy-4-chloroacetanilide, 10% 4-chloroacetanilide, and 8% 4 chloronitrobenzene. Its spontaneous decomposition in water also followed second order kinetics, K = 350 l mol-1 sec-1 and yielded N-(2-acetylamino-5 chlorophenyl)-p-benzo-quinoneimine-N-oxide in addition. 4. In the coupled oxidation of 10(-3) M haemoglobin-Fe2+ with 10(-3) M (II), 75% HbFe3+ was formed after 1 h, but only one third of the equivalent of oxygen was released, and two thirds were reduced to water. Concentration of (II) decreased by 5% only, indicating that one mol of (II) had catalysed the oxidation of 15 equivalents of haemoglobin-Fe2+. The identity of the product pattern formed with HbO2 with that produced by chemical one-electron oxidation indicated that oxygen bound to haemoglobin also functions as an acceptor for electrons from (II) as from (I), but the different redox potentials can explain why the secondary aromatic nitroxide was catalytically active and the primary nitroxide was not. PMID- 2546326 TI - Effect of 7-hydroxymethyl-12-methylbenz[a]anthracene and 1,3-bis-(2-chloroethyl) 1-nitrosourea on enzyme activities and oxidation of glutathione in cultured rat adrenal cells. AB - 1. The activities of enzymes participating in the regeneration of reduced glutathione (GSH), and their subcellular distribution were studied in cultured rat adrenal cells. 2. It has previously been shown that the adrenocorticolytic agent 7-hydroxymethyl-12-methylbenz[a]anthracene (7-hydroxymethyl-12-MBA) causes a drastic and selective oxidation of mitochondrial GSH in rat adrenal cells. Treatment of the adrenal cells with 7-hydroxymethyl-12-MBA, resulted in a minor decrease in the content of cytochrome c oxidase, nicotinamide nucleotide transhydrogenase, isocitrate dehydrogenase and cytosolic GSH reductase, whereas the activity of lactate dehydrogenase and citrate synthase was unaffected. None of these effects were considered to be responsible for the massive oxidation of mitochondrial GSH induced by 7-hydroxymethyl-12-MBA. 3. 1,3-Bis-(2-chloroethyl)-1 nitrosourea (BCNU) was used to obtain rat adrenal cells cultures with inactivated cytosolic and mitochondrial GSH reductase. The oxidation of mitochondrial GSH, induced by 7-hydroxymethyl-12-MBA, was not dramatically enhanced by the inactivation of GSH reductase, indicating that this enzyme was not rate-limiting in the regeneration of GSH. 4. Fractionation of rat adrenal cells with increasing concentrations of digitonin resulted in an earlier release of citrate synthase in cells treated with 7-hydroxymethyl-12-MBA compared with controls. These results may indicate damage to mitochondrial membranes as a result of 7-hydroxymethyl-12 MBA treatment. PMID- 2546329 TI - Immunogenicity of herpes simplex virus glycoprotein gB-1-related protein produced in yeast. AB - A protein related to glycoprotein B of herpes simplex virus type 1 (HSV-1) produced in yeast (ygB-1) was purified with an immunoadsorbent. The molecular weight of the purified ygB-1 as determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis was 96,000. Mice injected twice with ygB-1 adsorbed to alum developed ELISA antibody to ygB-1, neutralizing antibody to HSV 1 and a lymphoproliferative response to ygB-1 and HSV-1. The immunized mice were protected against intraperitoneal and corneal challenge with HSV-1. Latent infection in the trigeminal ganglia after corneal challenge was also inhibited by immunization with ygB-1. Guinea-pigs pigs immunized with ygB-1 adsorbed to alum also developed ELISA antibody to to ygB-1 and neutralizing antibody to both types of HSV. After the second dose, strong lymphoproliferative responses were seen upon stimulation with HSV-2. Animals were protected against intravaginal challenge with HSV type 2. PMID- 2546330 TI - Dual loci of DNA bending in the bovine papillomavirus upstream regulatory region. AB - Recombinant clones containing all or portions of the bovine papillomavirus (BPV) upstream regulatory region (URR) were constructed. When analyzed by polyacrylamide gel electrophoresis, the cloned URR sequences exhibited electrophoretic properties characteristic of DNA containing bend sites. Two distinct bend centers were identified, mapping to approximately 7477 and 7790 respectively on the BPV genome. No other loci of static DNA bending were detected in the URR region of the BPV genome. PMID- 2546328 TI - Intranasal immunization with proteoliposomes protects against influenza. AB - Worldwide, influenza virus remains a serious disease which has successfully eluded numerous attempts to design a consistently effective vaccine. In part, these attempts have been thwarted because of a lack of basic understanding of the mechanisms which mediate protection and recovery from influenza infection. A better understanding of the roles of secretory antibody, serum antibody and cell mediated immunity vis-a-vis protection and recovery from influenza infection has allowed us more rationally to approach the design and administration of a vaccine for influenza. We have constructed a vaccine composed of glycoproteins from the envelopes of either influenza of Sendai virus embedded in a lipid bilayer (immunosomes) mimicking the presentation of the virus to the cells during natural infection. Intranasal immunization with these immunosomes induces an adequate systemic Ir compared with intramuscular immunization and a superior local IgA response. These animals were specifically protected from virus challenge. PMID- 2546331 TI - Comparative features of a coronavirus isolated from a cheetah with feline infectious peritonitis. AB - A coronavirus which was isolated from a cheetah (Acinonyx jubatus) that succumbed to feline infectious peritonitis was characterized in vitro. The virus was determined to be highly cell-associated with Crandell feline kidney (CrFK) cells and was routinely maintained as a persistent infection (CrFK 83-4497). The cheetah coronavirus was compared with other members of the feline coronavirus group including the feline enteric coronavirus (FECV) 79-1683 and the feline infectious peritonitis viruses (FIPV), 79-1146, and UCD-1. The cheetah coronavirus was demonstrated to have a restricted host-cell range with limited cytopathic effect. Indirect immunofluorescence with antisera to FIPV UCD-1 revealed the concentration of viral antigens in the perinuclear region of cells infected with the cheetah coronavirus. Ultrastructural studies of the cheetah coronavirus indicated a limited number of immature viral particles within cytoplasmic vesicles and at the cell surface. This was in contrast to electron microscopy results of FECV 79-1683 and FIPV 79-1146, which had numerous mature virus particles within the cytoplasmic vesicles, as well as at the cell surface. The cheetah coronavirus was tentatively placed in the feline coronavirus family based upon its antigenic reactivity by immunofluorescence; however, the possibility that it represents a unique coronavirus of cheetahs should not be dismissed without further analyses at the host and genomic levels. PMID- 2546332 TI - The effect of E1 mutations on biochemical transformation by an adenovirus carrying the herpes simplex virus thymidine kinase gene in region E3. AB - A series of human adenovirus type 5 (Ad5) vectors has been constructed in which a vector containing the human herpes simplex virus thymidine kinase (TK) gene has been recombined with several Ad5 early region 1 (E1) mutants. The resulting viruses were used to study host-virus interactions in TK- rat cells and to examine the importance of E1 functions in a biochemical transformation assay. One of the most important parameters affecting transformation efficiency in this system was the cytotoxicity of the transforming virus. Ad5 viruses expressing the E1a 289 amino acid protein were all highly cytotoxic and induced significantly fewer colonies than did less cytotoxic mutants which were defective in expression of the 289 amino acid product. When correction was made for differential cell viability the variation in transformation efficiencies was considerably reduced although some E1a mutants still demonstrated an enhanced ability to transform in comparison to wt virus. The significance of these results to morphological transformation by adenoviruses is discussed. PMID- 2546334 TI - [Molecular mechanisms of neuronal plasticity during learning: the role of second messengers]. PMID- 2546333 TI - Change of processing and nucleocytoplasmic transport of mRNA in HSV-1-infected cells. AB - A monoclonal antibody (MAb) raised against the pore-complex lamina fraction from CV-1 cells was used to study alterations of gene expression in herpes simplex virus type 1 (HSV-1)-infected cells. This MAb, which recognized only one cellular polypeptide of 60,000 Da, selectively stained the nucleus in immunofluorescence microscopy, showing a punctuated pattern either at the nuclear surface or at the nuclear rim. By immunoelectron microscopy, the p60 antigen could be localized in the nuclear pore complex structure. Infection of CV-1 cells with HSV-1 resulted in a drastic change of the nuclear staining pattern. Four hours p.i., a clustering of the p60 antigen and, 12 h p.i., a formation of finger-like holes, penetrating the nucleus, occurred. Later in infection (22 h p.i.) the antigen was found to be almost absent. By RNA blot hybridization it was demonstrated that, after HSV-1 infection, the level of cellular mRNA (beta-tubulin) gradually decreased, while the level of HSV major DNA binding protein (DBP) mRNA increased, reaching maximal level 3-6 h p.i. Interestingly, the level of beta-tubulin gene transcripts changed differentially in the polysomal and in the nuclear fraction during the initial phase of infection, in contrast to the viral DBP transcripts, indicating that, after HSV infection, host cell transcripts accumulate in the nucleus. Evidence is presented indicating that this change is not due to altered nucleocytoplasmic mRNA transport but is due to an impaired splicing of host cell transcripts in HSV-infected cells. The MAb, directed against the nuclear pore p60 antigen, strongly inhibited the ATP-dependent efflux of both cellular and viral mRNA from isolated nuclei. The ATP-dependence of the efflux did not change during viral infection. However, the inhibitory potency of the MAb was found to be lost at the final stage of HSV infection, paralleling the loss of p60 antigen. PMID- 2546335 TI - Biochemical analysis in the yeast genus Candida. PMID- 2546336 TI - The glucose-induced CDC25- and RAS-mediated cAMP signal in the yeast Saccharomyces cerevisiae. PMID- 2546337 TI - Increase in cyclic AMP content with enhanced phosphatidylinositol turnover in the cells of Candida tropicalis during mycelial growth caused by ethanol. AB - Ethanol causes mycelial growth of Candida tropicalis Pk 233, which is associated with enhanced metabolism of phosphatidylinositol at the mid-log phase of growth, and the effects of ethanol are prevented by concomitant addition of myo-inositol (FEBS Lett. 214, 127-129, 1987). Ethanol induced also a marked increase in cellular content of cAMP at the mid-log phase, and myo-inositol abolished this effect of ethanol. The elevated level of cAMP content caused by ethanol was gradually lowered through the late-log and stationary phases and reached to control level. Very similar effects of ethanol and myo-inositol were observed in adenylate cyclase activity, while the activity of cAMP phosphodiesterase was not affected by ethanol. The ethanol-induced change in cAMP content was therefore ascribed to that in adenylate cyclase activity. These results suggested that cAMP plays an important role in combination with phosphatidylinositol turnover in the development of mycelial form in this dimorphic yeast. PMID- 2546339 TI - Evaluation of an enzyme-linked immunosorbent assay for detection of antibodies to bovine virus diarrhoea virus in milk. AB - The present study shows that milk is an appropriate source for detection of seroreactors to bovine virus diarrhoea virus (BVDV). There was close agreement between antibody titres in serum and in skim milk, as determined by an indirect enzyme-linked immunosorbent assay. The antibody titres were usually lower in skim milk than in serum, but all seropositive cows (n = 84) were also skim milk positive and all but one seronegative cow (n = 55) proved negative in skim milk. During lactation, the level of antibodies to BVDV in milk showed an inverse relationship to the amount of milk produced. However, there was a sufficient level of antibodies in milk throughout lactation to permit an adequate determination of BVDV antibody status in dairy cows. There was a mutual good agreement between milk antibody titre in the four mammary quarters, irrespective of milk cell count. Milk can be used to detect seroreactors to BVDV. Milk is preferable to blood in large-scale epidemiological studies, since the sampling procedure is much simpler. PMID- 2546338 TI - [Resection of lung metastases of colorectal cancer. Indications and indication limits]. AB - From 1970 through 1986, a total of 62 patients underwent thoracotomy for colorectal metastases to the lungs. Four had exploration only, whereas in 13 a nonradical, and in 45 a "curative" resection was performed. There was one postoperative death. Cumulative 5-year survival following radical resection was 44%. Conversely, all other patients succumbed within 3 years. Extrapulmonary disease has not yet been associated with 5-year survival and may usually contraindicate resection. The impacts of multiple metastases exceeding 3 nodules or bilateral pulmonary involvement on prognosis can not been determined from our series, since the procedure in respective patients was not radical in the majority of cases. In turn, localisation and venous drainage of the primary tumor did not significantly influence long term survival. PMID- 2546340 TI - Humoral immunity of neonatal swine after FMD vaccination. AB - The possible effect on pig protection after vaccination pregnant sows and their piglets against FMD at various age was examined using the SN test. Three experiments were conducted with three sow in each (8-9 piglets each). In the first experiment sows were not vaccinated but their piglets were vaccinated on 10th, 20th and 60th day of age. In second experiment sows were vaccinated at the end of rest period and in the middle of pregnancy. Pigs from one sow were vaccinated on 10th, from second sow on 20th and from the third sow on the 60th day of age. Pigs which were vaccinated on the 10th and 20th day of age were revaccinated on the 60th day of age. In third experiment sows were vaccinated at the end of the rest period and in the middle of pregnancy. Their piglets were not vaccinated. The conclusion may be drawn that if the epidemiological situation requires, systemic vaccination of all pigs then, on large farms (where pig production is planned) all sows should be vaccinated at the end of the rest period (first vaccination) and on the 55th-60th day of pregnancy. The next vaccination should be done in the middle of the next pregnancy. A vaccination program of piglets, according to our results, should depend on the vaccination program used for their dams. PMID- 2546341 TI - Coronavirus as agent of neonatal calf diarrhea in southern Chile. AB - Neonatal calf diarrhea is one of the most serious problems of cattle industry all over the world. Although the aetiology is complex, it is possible to assess that viruses play a very important role. During an investigation to study the importance of Rotavirus in enteric problems in calves and piglets, it was possible to demonstrate the presence, for the first time in southern Chile, of Coronavirus-like particles in a faecal sample of a 15 day old calf by means of electron microscopy. The importance of this diagnosis and some clinical characteristics of the disease are briefly discussed. Finally, the need for more research on this complex subject is recognized. PMID- 2546342 TI - Antitumor effect of adriamycin entrapped in liposomes conjugated with anti-bovine tumor antigen monoclonal antibody in leukemic cows. AB - Monoclonal antibody c143 against tumor-associated antigen (TAA) expressed on bovine leukemia cells was conjugated to liposomes containing adriamycin (ADM), and therapeutic effects of conjugates were examined in leukemic or preleukemic cows to prevent the progression of the disease. Five cows with TAA-positive in their peripheral blood lymphocytes were divided into two groups. Each group of cows received 4 injections of ADM alone (0.4 mg/kg) or c143-conjugated liposomes containing the same dose of ADM (L-AMD-c143) through the jugular vein at about 4 day intervals. In three animals treated with L-ADM-c143, the TAA-positive cells gradually decreased with treatment and finally two animals became TAA-negative during a 6 week period and a 14 week period after treatment, respectively. In the control, two animals treated with ADM alone showed only a slight decrease of TAA positive cells. PMID- 2546343 TI - [Various debated questions of the functional designation of phosphagene kinase]. PMID- 2546344 TI - [Functional blocks of the signal transmission system in retinal photoreceptors: evolutionary aspects]. PMID- 2546345 TI - [Functional organization and evolution of hormone-competent systems]. PMID- 2546346 TI - [Correlation between motor terminal latency time of the median nerve and the result of conservative treatment (cortisone injection) in carpal tunnel syndrome]. AB - 45 patients with carpal tunnel syndrome could be followed after an initial conservative treatment (local injection with Depomedrol 40 mg) either up to a surgical intervention (15 patients) or up to 14 to 48 months after the first injection (mean 23 months). Pregnant patients were excluded, as well as the patients with a distal motor latency (DML) greater than or equal to 7 msec (immediate surgical intervention). The age and sex distribution corresponds with the data reported in the literature: 7 males (15%) versus 38 females (85%), including 7 between 20 and 39 years (16%), 24 between 40 en 59 years (53%) and 14 between 60 and 80 years (31%). The side of the complaints also corresponds with the literature data: bilateral 16 (36%), right hand 23 (51%), left hand 6 (13%). In the 16 patients with a DML greater than 5,5 msec (normal value up to 4.3 msec) a rapid recurrence of the complaints after the injections necessitated a surgical intervention in 12 cases. In the 29 patients with a DML less than or equal to 5.5 msec who had been operated, the improvement of the complaints after the first injection lasted 2 months at most in 8 cases, as opposed to an average of 12.5 months (0 to 48 months) for the population as a whole. PMID- 2546347 TI - [Myoblastic myoma of the tendon of long flexor of the thumb]. AB - The case history of a female patient with flexion contracture of the thumb, having been caused by a tumour originated in the tendon of musculus flexor pollicis longus and histologically classified as a myoblastic myoma is presented. After a complete resection of the tumour and substitution of the originated defect of the tendon the function of the right hand became possible to a full extent and no signs of a relapse may be seen four years after the operation. PMID- 2546348 TI - Cervical cancer screening and demonstration projects to identify barriers to preventing cervical cancer mortality. AB - Cervical cancer screening and demonstration projects to identify barriers to optimal screening are discussed. Interview surveys showed that older women and women in low-income groups tended to have lower rates of Papanicolaou smear screening. Data produced by demonstration projects established by the Centers for Disease Control in collaboration with state and local authorities and private institutions will be used to design and implement strategies for increasing screening levels to further reduce cervical cancer mortality. PMID- 2546349 TI - Experience in managing a large-scale rescreening of Papanicolaou smears and the pros and cons of measuring proficiency with visual and written examinations. AB - Experiences in a large-scale interlaboratory rescreening of Papanicolaou smears are detailed, and the pros and cons of measuring proficiency in cytology are discussed. Despite the additional work of the rescreening project and some psychological and technical problems, it proved to be a useful measure of the laboratory's performance as a whole. One problem to be avoided in future similar studies is the creation of too many diagnostic categories. Individual testing and certification have been shown to be accurate predictors of proficiency. For cytology, such tests require a strong visual component to test interpretation and judgment skills, such as by the use of glass slides or photomicrographs. The potential of interactive videodisc technology for facilitating cytopathologic teaching and assessment is discussed. PMID- 2546350 TI - Canadian experience in cytology proficiency testing. AB - The mandatory Laboratory Proficiency Testing Program (LPTP) of the province of Ontario as applied to cytology laboratories is briefly described. LPTP evaluates reporting of test slides to identify laboratories that have deficiencies. Such laboratories receive an on-site visit, followed by recommendations on means of improvement (especially educational) and subsequent monitoring. Most of the 18 cytology laboratories visited to date have shown an improved level of performance on later tests. PMID- 2546351 TI - Epilogue. Establishment of the Center for Chronic Disease Prevention and Health Promotion. AB - The establishment of the Center for Chronic Disease Prevention and Health Promotion (CCDPHP) at the Centers for Disease Control (CDC) following the Conferences on the State of the Art in Quality Control Measures for Diagnostic Cytology Laboratories is briefly discussed. The CCDPHP is expected to play a major role in the CDC's cancer control program, including participation in establishing effective screening programs and assuring the quality of such methodologies as the Papanicolaou test and mammography. PMID- 2546353 TI - Presence and distribution of LHRH- and alpha-MSH-like immunoreactive nerve fibers in the epithalamus of the sheep. AB - The presence of luteinizing-hormone-releasing hormone (LHRH)-like and alpha melanocyte-stimulating hormone (alpha-MSH)-like immunoreactive fibers, nerve terminals or cellular elements in the pineal gland of the sheep has been investigated by immunohistochemistry. No LHRH-or alpha-MSH-like immunoreactive fiber, nerve terminals or cellular elements have been demonstrated in the pineal organ of the sheep. However, LHRH- and alpha-MSH-like immunoreactive fibers are present in the posterior commissure. Immunoreactive LHRH and alpha-MSH nerve endings are evident in the medial and lateral habenular nuclei. Discrepancies with the results obtained in other mammals are indicative of species differences in the distribution of LHRH- and alpha-MSH-like immunoreactive material in the pineal region. PMID- 2546352 TI - Bronchoalveolar lavage in liver transplant patients. AB - Because immunosuppression is required to control rejection, liver allograft recipients are susceptible to a variety of opportunistic pathogens. A total of 191 bronchoalveolar lavage (BAL) specimens from 89 patients (53 adults and 36 children) who underwent orthotopic liver transplantation was reviewed. One case each of cytomegalovirus (CMV), staphylococcal and Enterobacter pneumonia was diagnosed with the aid of pretransplant BAL. The pretransplant BAL in 62 patients showed rare yeasts in 24.2%; these probably represent oropharyngeal contaminants since the patients involved had no symptoms of Candida pneumonia. Among 54 patients who developed respiratory symptoms and underwent posttransplant BAL, 23 (42.6%) were infected with opportunistic pathogens, including Pneumocystis carinii (22.2%), CMV (22.2%) and herpes simplex virus (HSV) (7.4%). Frequently, infection with multiple organisms was present. Adults constituted 100% of the HSV infected group, 69.2% of the CMV-infected group and 16.6% of the group infected with P carinii. The diagnosis of these infections was aided by a combination of cytology, microbial culture and in situ hybridization techniques. Although BAL permitted the diagnosis and treatment of opportunistic infections, high mortality (62.5%) occurred with CMV and HSV pneumonia. Further studies into methods that permit earlier diagnoses of these infections are necessary. PMID- 2546354 TI - Nerve growth factor receptor immunoreactivity in human benign peripheral nerve sheath tumor. AB - In situ expression of nerve growth factor (NGF) receptors in human dermal and plexiform neurofibroma, schwannoma and traumatic neuroma was examined by an immunohistochemical method using a monoclonal anti-human NGF receptor antibody. Immunoreactivity for the NGF receptor was observed on the principal cells of both neurofibroma and schwannoma. Immunostaining by the anti-S-100 beta protein antibody in adjacent sections suggested that the vast majority of NGF receptor positive cells were also positive for S-100 beta protein. In traumatic neuroma, staining for the NGF receptor was more intense in the perineurium than in the endoneurial cells. PMID- 2546355 TI - A new type of neuronal cytoplasmic inclusion: histological, ultrastructural, and immunocytochemical studies. AB - A novel type of non-viral cytoplasmic inclusion is described, which was seen in virtually every neuron in the brain and spinal cord of a child with a presumed metabolic disorder whose clinical picture and CNS pathology were compatible with Leigh Syndrome. The ovoid to round inclusions were sharply demarcated, measuring up to 11 microns in diameter. They showed no distinctive staining with a battery of routine histological techniques. The ultrastructural features are unique, comprising non-membrane-bounded aggregates of randomly oriented plate-like structures with parallel linear densities depicting a periodicity of 11-16 nm. Immunocytochemical studies revealed strong staining with antisera to tropomyosin and weaker staining with antisera to actin. There was no reactivity with antibodies against neurofilaments, microtubules and their associated proteins, paired helical filaments, ubiquitin, vinculin or alpha-actinin. It is postulated that the metabolic disorder resulted in a neurodegenerative condition which manifested pathologically with lesions compatible with those of Leigh Syndrome. Associated with the condition was the discrete accumulation of cytoplasmic proteinaceous components, including tropomyosin, in the form of neuronal cytoplasmic inclusions possibly resulting from an alteration of the neuronal cytoskeleton. PMID- 2546357 TI - Malignant melanotic neuroectodermal tumor arising from the pineal body. AB - A case of a melanotic neuroectodermal tumor arising from pineal region of a 4 year-old girl is presented. The tumor had spread diffusely to the meninges, consistent with malignant behavior. Histologically, the tumor consisted primarily of epithelial elements arranged in tubules, cords and nests separated by fibrous vascular tissue in addition to a small neuroblastomatous focus. Melanin pigment was frequently observed in the epithelial tumor cells, and melanin-laden macrophages were also often observed. No teratoid elements were found. Immunohistochemically, tumor cells were positive for neuron-specific enolase but were nonreactive for S-100 protein, epithelial membrane antigen, glial fibrillary acidic protein, vimentin, alpha-fetoprotein and human chorionic gonadotrophin. Ultrastructurally, the epithelial nature of the tumor cells could be easily demonstrated. In addition, melanosomes in various stages in maturation were observed, indicating melanogenesis of the tumor. On the basis of the tumor location and the histological similarities previously observed for the fetal pineal body, it is very likely that this melanotic epithelial tumor could have originated from the fetal pineal gland. PMID- 2546356 TI - Polyglucosan bodies in intramuscular motor nerves. AB - The presence of polyglucosan bodies was studied in intramuscular motor nerves of 292 muscle biopsies. These biopsies were classified into five diagnostic categories and investigated for the presence of polyglucosan bodies in relation to age and sex. Their presence was nonspecific in patients over 20 years, the only correlation being with ageing. Under 20 years, their presence pointed to the diagnosis of Lafora's disease. In cases in which both a muscle biopsy and a sural nerve biopsy were performed, the former appeared to contain these polyglucosans more frequently. PMID- 2546358 TI - Distribution and morphology of brain stem plaques in Alzheimer's disease. AB - The morphology, incidence and distribution of senile plaques in the brain stem were examined in 15 cases of Alzheimer's disease, using mainly the Methenamine Bodian method. The plaques were found in all cases and were grouped into three types according to their morphology. They were not randomly scattered in the brain stem, but had a distribution common to all cases. There were numerous plaques in the periaqueductal gray, superior colliculus, fourth-ventricle floor and superior central nucleus. The plaques were also found less abundantly in the reticular formation, substantia nigra, pontine nucleus and inferior olivary nucleus. There was a tendency for certain plaque types to be associated with specific regions. In the familial cases, the plaques tended to occur even in the regions where they were rare in other cases. The capillaries with plaque-like degeneration were observed not infrequently in the brain stem. The distribution of plaques did not always coincide with that of neurofibrillary tangles. PMID- 2546360 TI - Ocular blood flow. Proceedings of the Oxford symposium. July, 1988. PMID- 2546359 TI - A neuropathological subset of Alzheimer's disease with concomitant Lewy body disease and spongiform change. AB - The neuropathological heterogeneity of Alzheimer's disease (AD) is increasingly recognized. Diffuse Lewy body disease, for example, most frequently occurs in cases fulfilling histopathological criteria for AD, and these patients usually present with dementia rather than parkinsonism. We report five cases of concomitant AD and diffuse Lewy body disease with still another coexistent neuropathological feature: localized and stereotyped spongiform change in the neuropil. This spongiform change was most striking in the superior and inferior temporal, entorhinal, and insular cortex and the amygdala and was virtually indistinguishable from that seen in Creutzfeldt-Jakob disease. Electron microscopic study on one case revealed membrane-containing vacuoles in close association with neuritic plaques and plaired helical filament-filled processes. Immunocytochemistry using antibodies to prion proteins (PrPsc or PrP27-30) failed to label plaque or vascular amyloid in the five cases. Four primates inoculated with brain tissue from one case have not evidenced neurological disease in the 3 years since the transmission experiment. We conclude that these cases represent a neuropathological subset of AD with relatively widespread Lewy bodies and a localized spongiform change, predominantly involving the medial temporal region. Despite the light and electron microscopic commonality with Creutzfeldt-Jakob disease, there is no clear evidence that these cases represent a form of transmissible spongiform encephalopathy. PMID- 2546361 TI - Ocular and retinal blood flow. AB - This paper discusses the interrelationship between retinal and choroidal blood flow, citing particularly the circulatory changes in light and dark adaptation, and the response vessels to hypoxia. The problems of diabetic retinopathy are also considered. PMID- 2546362 TI - Ocular blood flow changes with increased vascular resistance external and internal to the eye. AB - Precise instantaneous digital measurement of intraocular pressure done non invasively is a clinical window to the vascular physiology of the eye. This technique has been used to study the pulse amplitude/intraocular pressure curve in 2 distinct clinical entities. The first is amaurosis fugax before and after carotid endarterectomy. This demonstrates the effects of high vascular resistance external to the eye on the ocular blood flow. The second is a patient with long standing arteriosclerotic cardiovascular disease. This is an illustration of increased vascular resistance within the eye. PMID- 2546363 TI - Estimation of pulsatile ocular blood flow from intraocular pressure. AB - A relationship has been derived between intraocular pressure and pulsatile blood flow in the eye. Measurements of intraocular pressure show a time variation that is associated with the pulsatile component of arterial pressure. Experimental results provide a means of transforming intraocular pressure changes into ocular volume changes. The eye is represented by a chamber with elastic walls, a pulsatile incoming flow of incompressible fluid (blood), and a steady outgoing flow of blood. Under these conditions, the rate of pulsatile blood flow through the eye can be approximated from the instantaneous intraocular pressure measurements. Data from a healthy human eye are used to illustrate the analysis. PMID- 2546364 TI - Disc haemorrhages and the etiology of glaucoma. AB - As a consequence of the hypothesis that all open angle glaucomas at some time exhibit a disc haemorrhage, it is proposed that both low and high tension glaucomas have the same etiology. That the primary lesion should be sought in the small vessels of the optic disc seems to be a reasonable suggestion. PMID- 2546365 TI - Loss of the ocular pulse in the acute phase of temporal arteritis. AB - Severe cases of temporal arteritis often have infarctions of the optic nervehead. Such cases frequently have a decrease in the amplitude of the ocular pulse. This is in contrast to cases of anterior ischemic optic neuropathy not due to temporal arteritis. In addition to aiding in the diagnosis of temporal arteritis, monitoring of the ocular pulse may help in following the course of this disease. PMID- 2546366 TI - Cataract surgery and vascular changes in the eye. PMID- 2546367 TI - Ocular haemodynamics in cataractous eyes. A pilot study. AB - Pulsatile ocular blood flow, systolic and diastolic intraocular pressures and ocular perfusion pressures were measured in a group of 10 patients (mean age 67 +/- 3 years) with bilateral senile cataract. Each pair of eyes comprised one phakic and one pseudophakic eye. The results are compared to a group of 8 subjects (mean age 65 +/- 4 years) with no apparent ocular pathology. The mean ocular pulsatile blood flow of 394 +/- 52 microliters min-1 in the cataractous eyes was significantly reduced P less than 0.001) compared to that of the control group (571 +/- 40 microliters min-1). Results were similar for undisturbed cataractous eyes (mean pulsatile blood flow = 400 +/- 54 microliters min-1) and pseudophakic eyes (mean pulsatile blood flow = 388 +/- 51 microliters min-1). The intraocular pressures were within normal limits (systolic intraocular pressure = 19.7 +/- 0.7 mmHg, diastolic intraocular pressure = 17.5 +/- 0.6 mmHg) as were the ocular perfusion pressures (73.7 +/- 0.5 mmHg). The results provide evidence of decreased pulsatile ocular blood flow and associated ischaemia in cataractous eyes. PMID- 2546368 TI - Blood flow in the human eye. PMID- 2546369 TI - Use of continuous subcutaneous growth hormone-releasing hormone (GHRH (1-29)NH2) infusions to augment growth hormone secretion and to promote growth. AB - It has previously been shown that an 8-day continuous subcutaneous infusion of GHRH (1-29)NH2 in adult males augments growth hormone (GH) secretion with no evidence of desensitization of the response. The present report details the results of treatment with continuous subcutaneous infusions of GHRH (1-29)NH2 in eight children aged 6-9 years with partial GH insufficiency. All the children showed augmentation of GH secretion and an increase in growth velocity after 3 and 6 months of therapy. PMID- 2546370 TI - Modification of 24-hour growth hormone secretion after continuous subcutaneous infusion of growth hormone-releasing hormone (GHRH (1-29)NH2) in short children with low 24-hour growth hormone secretion. AB - Six short children with low 24-hour growth hormone (GH) secretion were treated with continuous subcutaneous infusion of GHRH (1-29)NH2 for 3 weeks using a portable infusion pump. Restoration of pulsatile GH secretion was observed in all three children treated with 40 micrograms/kg/day of GHRH, but in only one of the three children treated with 20 micrograms/kg/day. All parameters of 24-hour GH secretion increased, but in five children the magnitude of the GH response was greater on day 1 than on day 21 of GHRH treatment. This decrease was not observed in the single child who responded to a low dose of GHRH (20 micrograms/kg/day); on the contrary, the response in this patient was greater after 21 days of GHRH treatment. Plasma levels of GHRH (1-29)NH2 were significantly higher on day 21 than on day 1 of treatment, suggesting altered pharmacokinetics over time. The effect of GHRH treatment on growth could not be determined because of the short duration of the study, but the data obtained on 24-hour GH secretion and GHRH metabolism suggest that a long-acting analogue of GHRH could be useful for the treatment of GH deficient or insufficient children. PMID- 2546371 TI - Aldosterone secretion during acute respiratory acidosis and NH4Cl-induced metabolic acidosis in the goat. AB - Acute respiratory acidosis was induced in goats by inhalation of 6% or 8% CO2 in air for 30 min. The lower CO2 concentration caused a significant rise in plasma cortisol (PC), but had no appreciable influence upon plasma aldosterone (PA), and did not affect the arterial blood pressure (aBP). A more pronounced PC response was observed in association with the inhalation of 8% CO2, but also here without concomitant increase in PA. However, the aBP became elevated by about 30% during the CO2 exposure with a simultaneous increase in glomerular filtration rate and a water diuresis, suggesting that the release of arginine vasopressin temporarily became inhibited. It was confirmed that metabolic acidosis induced by duodenal NH4Cl administration is preceded by a transient rise in PA. Dexamethasone-induced feedback inhibition of the ACTH secretion blocked the PA response, which possibly reflects NH4 ion stimulation of the ACTH release. The combined results of the CO2 and NH4Cl experiments seem to justify the conclusion that increases in PA seen in conjunction with acidosis do not reflect a direct hydrogen ion stimulation of the adrenal glomerulosa cells. PMID- 2546372 TI - Effects of digitoxin and lithium, used as a marker of passive Na transport, on secretin-dependent bile flow in the pig. AB - The present study was performed in anaesthetized pigs, and the first aim was to assess the role of Na,K-ATPase in secretin-dependent biliary HCO3 secretion (JbHCO3). Intra-arterial administration of the cardiac glycoside digitoxin (0.2 mg/kg-1) reduced hepatic Na K-ATPase activity, JbHCO3 and secretin-dependent bile flow by 24, 55 and 34% respectively. In the second part of this study lithium (Li) was used as a marker of passive Na transport to assess the electrochemical gradient for Na flux into bile duct lumen during secretin-stimulated bile flow and impeded biliary osmotic water flow by i.v. infusion of glucose. At plasma glucose 85 (73-96) mmol l-1, bile [Na] and [Li] exceeded their concentrations in plasma by 57 and 47% respectively. By using the Nernst equation, transepithelial potential difference (PD) during hyperglycaemia was estimated to be -6.2 (0 to 10.8) mV (ductal lumen negative), which corresponds to a [Li]bile/[Li]plasma ratio of 1.3 (1.0-1.5). The ratio was not significantly different from the observed [Li]bile/[Li]plasma ratio of 1.4 (1.3-1.5). It is concluded (1) that Na, K-ATPase is necessary for JbHCO3, probably by sustaining the cell membrane PD (cell interior negative) which is a driving force for apical electrogenic HCO3 secretion, and (2) transepithelial Li (and hence Na) flux is driven solely by the negative transcellular PD during secretin-stimulated bile flow in the pig. PMID- 2546373 TI - Mechanism of action of calcitonin on secretion in rat submandibular gland. AB - Calcitonin (CT) was found to reduce the initial flow of pilocarpine-stimulated saliva from the submandibular glands in the rat. Although there was a concomitant increase of the concentration of calcium and protein in the saliva, the calcium/protein ratio was not significantly affected. CT also caused a significant increase of the potassium concentration in submandibular saliva. Both in vivo and in vitro, CT inhibited the production of cyclic AMP (cAMP) both in the absence and in the presence of forskolin. This decrease in intracellular cAMP levels could result in an inhibition of mucus secretion, which would explain the previously observed calcitonin-induced intracellular accumulation of mucus in the submandibular gland acinar cells. CT did not affect the cytoplasmic free Ca2+ concentration (as measured with fura 2) in isolated submandibular acini either in the absence or in the presence of cholinergic or adrenergic agonists. These results indicate that the inhibition of fluid secretion in the submandibular gland by calcitonin must be located distal to changes in [Ca2+]i. It can be concluded that CT affects both mucus and fluid secretion in the submandibular gland, but that only the inhibition of mucus secretion can as yet be explained by an effect at the level of the second messenger. PMID- 2546374 TI - Excitatory amino acids and cerebrovascular tone. AB - Levels of excitatory amino acids in the brain extracellular fluid compartment rise during pathological conditions in the brain such as ischaemia, anoxia and epilepsy. One such amino acid, glutamate, is present in sensory nerve fibres innervating, for example, cerebral vessels. Enhanced levels of circulating glutamate and aspartate are found in migraine sufferers. The present study examined whether excitatory amino acids, in concentrations found in the brain extracellular fluid compartment during pathological conditions, exert a direct effect on cerebrovascular tone. As tested in flow-regulating pial arteries from rat, cat and man, no such constrictive or dilatory effect was obtained. PMID- 2546376 TI - Amphetamines: pharmacology, abuse and addiction. AB - Amphetamine (beta-phenylisopropylamine) is a potent sympathomimetic amine of a simple structure with a multiplicity of biological effects that include hyperthermic, anorectic, cardiovascular and central nervous system stimulant actions. Since the 1930s a large number of drugs have been developed from systematic, chemical modifications of the basic amphetamine molecule to emphasize some of the properties of amphetamines and to eliminate or diminish others. These chemical manipulations have resulted in the synthesis of a variety of more selectively acting sympathomimetics. These altered molecules include CNS stimulants, potent psychomimetics (hallucinogens), anorectic agents, and vasoconstrictors that all have the basic beta-phenylisopropylamine skeleton. Reports of the consequences of abuse and addiction followed rather closely the development of these agents: manufacture, distribution and use continue to the present day. Both legitimate and illicit production account for a significant level of use of CNS stimulants. CNS stimulants are perhaps the most reinforcing drugs known to man. For this reason alone they will persist as drugs of choice among a variety of personalities. PMID- 2546375 TI - Managing hypertension in family practice: a nationwide collaborative study of the use of four antihypertensives in the treatment of mild-to-moderate hypertension. A report from CEN. Clinical Experience Network. AB - The goals of this prospective, nonexperimental study were to examine the ways in which family physicians select from among four antihypertensive agents for their patients and to provide an overall perspective on how these agents perform in the management of hypertension in primary care. Three hundred seventy-eight family physicians treated 3608 mild and moderate hypertensives with one of the following medications: atenolol (n = 564 patients), enalapril maleate (n = 677), verapamil hydrochloride in sustained-release form (n = 1861), or a fixed combination, hydrochlorothiazide/triamterene (n = 506). The resultant four groups of patients differed in several demographic and clinical measures: age, gender, race, concurrent disease, diastolic and systolic blood pressures, heart rate, and history of hypertension. The patient profiles for each group suggest appropriate matching of drugs to individual patient needs: younger patients and those with higher heart rates more often received the beta-blocker; blacks were more frequently assigned to the diuretic and less often to the beta-blocker; patients with concurrent diseases and a longer history of hypertension were more often assigned to the angiotensin-converting-enzyme (ACE) inhibitor or the calcium channel blocker. Rates of success, defined by the percentages of patients staying on the selected drug and experiencing a reduction of at least 10 mmHg or achieving a diastolic pressure less than or equal to 90 mmHg, were in the same range for all four groups (55 to 62.5 percent). Patients evaluated their quality of life and gave enalapril and verapamil SR the highest ratings. The rapid completion of the study, the quality of the results, and the high rates of follow up and compliance show that family practice is an excellent setting for conducting clinical research. PMID- 2546377 TI - Molecular biology of leukotriene and lipoxin formation. PMID- 2546378 TI - Sequential formation of leukotriene E4 metabolites by isolated rat hepatocytes. PMID- 2546379 TI - Metabolism of leukotriene B4 and related substances by polymorphonuclear leukocytes. PMID- 2546380 TI - Enhancement of catecholamine release may be mediated by prostaglandin E receptor stimulated phosphoinositide metabolism. PMID- 2546381 TI - Co-expression of leukotriene B4 and leukotriene D4 receptors on human monocytic leukemia U-937 cells. PMID- 2546382 TI - Involvement of peptidoleukotrienes in antigen-dependent thromboxane (TX) synthesis in IgG1-sensitized guinea pig lungs. AB - Lungs from IgG1-sensitized guinea pigs synthesize both leukotrienes (LTs) and thromboxane (Tx) upon ex vivo antigen challenge. This study was undertaken to investigate whether antigen-dependent Tx synthesis could result from prior formation of LTD4. In IgG1-sensitized lungs, LTD4 effectively induced Tx formation (ED50 = 2-4 nM). In these lungs, the levels of antigen-dependent formation of LTD4 (8-26 nM formed by 0.01-10 micrograms/ml antigen challenge) were 2-7 X greater than the ED50 value of LTD4-stimulated Tx synthesis. In addition, incubation of the sensitized lungs with ICI-198,615, a LTD4 antagonist, prior to antigen-challenge prevented Tx formation (IC50 = 0.01 microM). Our results indicate that LTD4 generated from IgG1-sensitized lungs could play a prominent role in stimulating Tx synthesis. LTC4 may also be involved because of rapid formation of LTC4 upon antigen-stimulation and its capacity to induce Tx synthesis. PMID- 2546383 TI - Molecular diversity of human leukocyte receptors. PMID- 2546384 TI - Inhibition of in vitro guinea pig eosinophil chemotaxis by leukotriene B4 antagonist U-75,302. PMID- 2546385 TI - Synthesis of leukotrienes C4, D4, E4 and 20-carboxy-B4 by the isolated perfused kidney. PMID- 2546386 TI - Leukotriene D4 (LTD4)/leukotriene B4 (LTB4) interactions in the pathophysiology of experimental glomerulonephritis. PMID- 2546387 TI - Mechanism of action of adrenergic and cholinergic stimuli on cardiac prostaglandin synthesis. PMID- 2546388 TI - Corneal arachidonate metabolism via cytochrome P450: characterization of two novel biologically active metabolites. PMID- 2546389 TI - Recent development of platelet-activating factor antagonists. PMID- 2546390 TI - Chemistry and pharmacology of PAF antagonists. Evaluation of changes at potential metabolism sites on activity and duration of activity. PMID- 2546391 TI - Recent studies on icosanoids and PAF-acether modulation of some central neurosecretions in the rat. PMID- 2546393 TI - The use of leukotriene D4 receptor antagonists and 5-lipoxygenase inhibitors to define a role for leukotrienes in allergic reactions. PMID- 2546392 TI - Molecular mechanism of cyclooxygenase regulation in cloned mouse osteoblastic cell (MC3T3-E1). PMID- 2546394 TI - Beta adrenoceptor desensitization in lung: a role for prostaglandins. PMID- 2546395 TI - Role of cysteinyl leukotrienes in gastric mucosal damage in rats. PMID- 2546396 TI - Profile and sites of eicosanoid release in experimental necrotizing enterocolitis. PMID- 2546397 TI - Depletion of resident glomerular macrophages by essential fatty acid deficiency protects against glomerulonephritis. PMID- 2546399 TI - The effects of a fish oil enriched diet on experimentally-induced and clinical asthma. PMID- 2546398 TI - Bradykinin-induced activation of phospholipase A2 is independent of the activation of polyphosphoinositide-hydrolyzing phospholipase C. PMID- 2546401 TI - Effect of oral administration of highly purified eicosapentaenoic acid and docosahexaenoic acid on platelet function and serum lipids in hyperlipidemic patients. PMID- 2546400 TI - The effect of highly purified eicosapentaenoic acid in patients with psoriasis. PMID- 2546403 TI - Structural analysis of sulfido-peptide leukotrienes: application to the design of potent and specific antagonists of leukotriene D4. PMID- 2546402 TI - Synthesis of methanoleukotriene A4 analogs and their pharmacological activities. PMID- 2546404 TI - A simple purification method for quantitation of eicosanoids, in biological specimens by gas chromatography/selected ion monitoring. PMID- 2546405 TI - 11,12-leukotriene A4: synthesis and metabolism. AB - 11(S),12(S)-oxido-5Z,7E,9E,14Z-eicosatetraenoic acid (11,12-LTA4) was chemically synthesized from 12-hydroperoxy-eicosatetraenoic acid (12-HPETE). 11,12-LTA4 methyl ester was nonenzymically hydrolyzed to at least four products. These products were identified to be epimers of 5,12(S)-dihydroxy-eicosatetraenoic acid methyl ester (5,12(S)-diHETE-Me) and epimers of 11,12-diHETE-Me. In addition to the above nonenzymic products, 11,12-LTA4 was converted to 11,12-LTC4 by the rat liver glutathione S-transferase, and to an isomer of 11,12-diHETE by homogenates of the guinea pig adrenal gland and liver. PMID- 2546406 TI - Histamine-mediated regulation of cAMP levels and inositol phosphate metabolism in isolated rabbit retina. AB - In pieces of the rabbit retina, histamine (HI) had no significant effect on cAMP accumulation; however, HI inhibited forskolin-evoked accumulation of the nucleotide. Mepyramine (H1-antagonist) was without effect, while cimetidine (H2 antagonist) antagonized the HI action. Dimaprit (H2-agonist) mimicked the HI action, however its action was not significantly affected by H2-antagonists. Since phosphodiesterase (PDE) inhibitors prevented the HI action it is suggested that HI-induced inhibition of the forskolin effect results from HI-induced activation of cAMP breakdown. The muscarinic agonist carbachol, and to a lesser extent HI, both increased [3H]inositol incorporation and intensified accumulation of [3H]inositol phosphates in the rabbit retina. PMID- 2546407 TI - The possible role of histamine-mediated cAMP formation as a link between H2 receptor stimulation and ultrastructural changes in guinea-pig oxyntic cells. AB - Guinea-pig oxyntic cell tubulin has been isolated and in vitro aggregation has been studied. The spontaneous assembly of isolated tubulin was significantly accelerated and increased by 1 mmol/l GTP. Histamine and forskolin increased tubulin polymerization only when detergent dispersed oxyntic cells or crude membranes were added. The forskolin response occurred very rapidly with an EC50 of approximately 30 mumol/l and did not require GTP. Histamine promoted tubulin aggregation with an EC50 of about 5 mumol/l only in the presence of GTP. Ranitidine completely inhibited the effects of histamine. From these data it is suggested that, in the oxyntic cell, histamine H2-receptor activated adenylate cyclase and the corresponding increase in cAMP play a role in eliciting characteristic ultrastructural changes by initiating formation of microtubules as a first step in the cascade of events leading to an increase in the secretory surface area. PMID- 2546408 TI - [Phosphoinositide turnover and Ca2+ mobilization during phagocytosis in cultured chick retinal pigment epithelial cells]. AB - Ca2+ mobilization and phosphoinositide turnover were examined during phagocytosis of latex particles in cultured chick retinal pigment epithelial cells (RPE cells). Ca2+ influx into cells and Ca2+ efflux from the cells were enhanced by about 1.5-fold and 3-fold compared to control cells, respectively. A high content of Ca2+ in RPE cells has been reported. Therefore, the Ca2+ efflux observed here may be a reflection of Ca2+ release into cytosol from the intracellular storage site(s). In [3H] inositol-prelabeled RPE cells, addition of latex particles elicited decreases in phosphatidylinositol 4,5-bisphosphate and phosphatidylinositol with the concomitant formation of inositol phosphates including inositol trisphosphate, indicating the hydrolysis of phosphoinositides by phospholipase C. These results suggest that phosphoinositide turnover may be closely coupled with Ca2+ mobilization during phagocytosis in RPE cells. PMID- 2546409 TI - [The effect of vasoactive intestinal polypeptide (VIP) on ocular inflammation in rabbit ocular tissue]. AB - Intravitreous injection of vasoactive intestinal polypeptide (VIP) induced ocular inflammation in albino rabbits. Protein amounts and cell numbers in aqueous humor were measured. The effect of VIP on the release of cyclic AMP (cAMP) and prostaglandin E2 (PGE2) from the isolated albino rabbit irides was investigated. Mydriatic response was obtained by intravitreous injection of 0.5 microgram or less VIP. 5 microgram or more of VIP induced miosis and breakdown of the blood aqueous barrier. Intravitreous injection of VIP induced on increase of cAMP in the aqueous humor dose-dependently and a slight increase of PGE2. VIP induced a dose-dependent increase of cAMP release and slightly inhibit PGE2 release from isolated irides. In this study, a large quantity of VIP was injected intravitreously induced breakdown of the blood aqueous barrier. The effect of VIP on isolated irides was to activate the release of cAMP and to inhibit that of PGE2. PMID- 2546410 TI - [A case of acute retinal necrosis syndrome caused by herpes simplex virus type 1]. AB - A case of bilateral acute retinal necrosis syndrome accompanied by viral meningitis in a 46 year-old male is reported. The characteristic scattered yellowish-white retinal exudates and retinal detachment appeared in both eyes within 2 weeks, and after treatment with intravenous acyclovir and gamma globulin, and encircling, scleral buckling, pars plana vitrectomy, 100% SF6 tamponade procedures, the retinal detachment was repaired and the yellowish-white retinal exudates disappeared. On virological examination, the complement fixation titer for herpes simplex virus (HSV) in serum increased more than eightfold in pair serum, and the quotient of antibody for HSV type 1 (HSV-1) IgG was high. HSV 1 was isolated and cultured from the subretinal fluid. From these results, it is concluded that HSV-1 infection was the cause of the meningitis and acute retinal necrosis in this case. PMID- 2546411 TI - Mixed mullerian sarcoma of the uterus: MR imaging findings. AB - MR images of seven patients with histologically documented mixed mullerian sarcoma were analyzed retrospectively to determine whether the scans showed findings that could suggest the diagnosis. Spin-echo T1- and T2-weighted sagittal and transverse images from either a 0.35- or a 1.5-T unit were available for each patient. MR image analysis included evaluation of tumor signal intensity on T1 and T2 images; tumor location, size, and extent; depth of myometrial invasion; and presence of pelvic metastases. In all seven patients, MR images showed a large endometrial mass deeply invading the myometrium or beyond. In addition, MR images showed intraperitoneal (two patients) and ovarian (one patient) metastases. Although the massiveness of the tumors on initial presentation may suggest the diagnosis of mixed mullerian sarcoma, the MR imaging findings are nonspecific and mimic invasive endometrial carcinoma. PMID- 2546412 TI - Sonographically guided percutaneous injection of ethanol for treatment of hepatocellular carcinoma: value of Gelfoam to mark the lesion. PMID- 2546413 TI - T1 and T2 measurements of meningiomas and neuromas before and after Gd-DTPA. AB - Seven patients with meningiomas and five patients with neuromas were examined with spin-echo sequences on a 0.35-T imaging system. Signal enhancement and relaxation rate increments with gadolinium-diethylenetriamine pentaacetic acid (Gd-DTPA) were evaluated, as well as relaxation rate contributions, indicators of Gd-DTPA accessibility to the tissue water. The average signal enhancement rate with Gd-DTPA was higher in neuromas than in meningiomas, 148% and 84%, respectively, but this difference was poorly appreciated on enhanced images because of similar average postcontrast T1 values in both tumors. However, the average T1 relaxation increment was almost twofold higher in neuromas than in meningiomas, 318% and 162%, respectively, mainly deriving from longer intrinsic T1 values in neuromas. Also, there was a 25% increase in the average T2 relaxation rate in neuromas after Gd-DTPA together with a higher T2 contribution by Gd-DTPA, while such effects were hardly discernible in meningiomas, suggestive of greater accessibility of Gd-DTPA to the tissue water in neuromas. By electron microscopy, endothelial fenestration and open gap junctions are commonly found in capillaries of both tumors, functioning as the routes into the extracellular space where the major contrast effect of Gd-DTPA can be expected. However, the open gap junctions are short and straight in neuromas, while they are tortuous and sinusoid in meningiomas. This may provide greater access for contrast material into relatively larger extracellular spaces in neuromas and may cause a greater T1 relaxation rate increment with Gd-DTPA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546415 TI - Instantaneous transmitral flow using Doppler and M-mode echocardiography: comparison with radionuclide ventriculography. AB - To improve the accuracy of Doppler echocardiographic indices of left ventricular filling, we derived two indices of instantaneous transmitral flow with the use of Doppler velocities and M-mode echocardiography. These indices were calculated from the product of pulsed Doppler mitral velocities and either the excursion of the anterior mitral leaflet or the separation of both mitral leaflets as measures of the changing mitral orifice area. The derived flow indices and the mitral velocities alone were compared to left ventricular filling as determined by radionuclide ventriculography in 24 patients. When compared as areas under the matched decile divisions of the derived filling sequences by linear regression analysis, the relationship for combined Doppler and M-mode versus radionuclide left ventricular filling was closer to the line of identity (slope = 0.98 and 0.94 using the anterior mitral leaflet and both mitral leaflets, respectively, both p = NS versus the line of identity) than was the relationship for mitral velocities alone versus radionuclide left ventricular filling (slope = 0.74, p less than 0.05 versus the line of identity). The instantaneous mitral volume flow indices more closely resemble the time course and shape of radionuclide left ventricular filling curves than do mitral velocities alone, and the application of these indices should assist the quantitative description by Doppler echocardiography of left ventricular filling. PMID- 2546414 TI - Stimulation of atrial natriuretic peptide and vasopressin during percutaneous transluminal aortic valvuloplasty. AB - The objective of this study was to determine the effects of transient aortic valve occlusion (balloon valvuloplasty) on vasoactive hormones in patients with heart failure. Plasma atrial natriuretic peptide, vasopressin, aldosterone, adrenocorticotropic hormone (ACTH), and plasma renin activity were measured before, immediately after, and 30 minutes and 18 to 24 hours following balloon inflation in 18 patients. Mean right atrial and pulmonary wedge pressures were 6 and 18 mm Hg before inflations, respectively, and were unchanged after balloon inflations (5 and 13 mm Hg, respectively). Systemic systolic/diastolic pressures were 139 +/- 8/65 +/- 4 mm Hg before occlusion, decreased to 47 +/- 5/34 +/- 3 mm Hg during occlusion, and returned to baseline after occlusions. Baseline atrial natriuretic peptide levels were 267 +/- 43 pg/ml and increased to 513 +/- 71 pg/ml after balloon inflations. Vasopressin levels before occlusion were 9.1 +/- 2.2 pg/ml and increased to 21.4 +/- 4.8 pg/ml after balloon inflations. Plasma renin activity was 5.4 +/- 1.4 ng/ml/hr before inflations and was not significantly changed after balloon inflations. No clinically significant changes in plasma sodium, potassium, creatinine, and osmolality were observed after the procedure. Aldosterone increased from 23 +/- 4 to 40 +/- 7 ng/dl 10 minutes after the last inflation. Plasma ACTH measured in seven patients with increased aldosterone was 28 +/- 8 pg/ml before and increased to 295 +/- 157 pg/ml 10 minutes after balloon inflations. The increases in natriuretic peptide and vasopressin were likely due to elevated intracardiac and decreased arterial pressures, respectively; they persisted in spite of no clinically significant changes in filling pressures 12 to 24 hours after the procedure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546416 TI - Serum lipid response to dietary cholesterol in subjects fed a low-fat, high-fiber diet. AB - To determine whether hyperresponse to dietary cholesterol occurs in individuals consuming a low-fat diet, a study was performed on 58 subjects whose plasma cholesterol had changed by greater than or equal to 5% when intakes of two and seven eggs per week were compared in a previous study. Here, the effect of eating nine, zero, and nine eggs per week was studied over 3 consecutive months. Plasma total, LDL, and HDL cholesterol did not differ significantly. There was no correlation between individuals' lipoprotein changes in this and the previous study. Consistent hyperresponse to moderate cholesterol intake is not apparent in people eating a low-fat, high-fiber diet. Reduction in dietary cholesterol below 400 mg/d produces no further substantial cholesterol lowering. PMID- 2546417 TI - High-fiber diets: influence on characteristics of cecal digesta including short chain fatty acid concentrations and pH. AB - The effects of two sources of dietary fiber on the characteristics of cecal contents were assessed directly with miniature swine cannulated to facilitate frequent collections of cecal digesta. The short-chain fatty acid (SCFA) concentrations increased and the pH decreased at the same time that meal constituents entered the cecum; PEG was used as a dietary marker. The bean diet resulted in higher concentrations of acetate and total SCFA in cecal digesta, lower concentrations of butyrate, a larger SCFA pool size, and a more acidic pH than did the bran diet. Thus, we conclude that SCFA concentrations and acidity of the digesta are directly related and that dietary fibers can affect simultaneously several variables that are implicated as factors influencing colonic health. PMID- 2546418 TI - Hepatoblastoma in an adult with metastasis to the ovaries. AB - A 19-year-old female had ascites, an enlarged liver, pelvic masses, and an alpha fetoprotein level of 397,000 micrograms/L with a normal beta-human chorionic gonadotropin serum level. Abdominal exploration revealed a large solitary liver mass with bilateral ovarian masses, and bilateral salpingo-oophorectomy and wedge liver biopsy were performed. The tumor was composed of cords, nests, and pseudorosettes of polyhedral cells with eosinophilic cytoplasm and nuclei with prominent nucleoli. The clinical, light microscopic, and immunocytochemical features indicate that the primary liver neoplasm and ovarian metastases were a purely epithelial hepatoblastoma. To the authors' knowledge, this is the first reported case of this type. PMID- 2546419 TI - Human papillomavirus DNA determination of anal condylomata, dysplasias, and squamous carcinomas with in situ hybridization. AB - Infection with types 6, 11, 16, and 18 of the human papillomavirus (HPV) is associated with condylomatous, dysplastic, or carcinomatous changes in the genital tract. Emerging evidence suggests that a similar series of lesions develops in the anal canal after exposure to the same HPV types. In situ hybridization was performed with the use of biotinylated DNA probes to HPV 6, 11, 16, and 18, so as to determine the frequency of HPV DNA in 45 perianal and/or anal condylomata, 6 anal intraepithelial neoplasias, and 13 anal squamous cell carcinomas. Of the 33 perianal and/or anal condylomata in which HPV DNA was detected, 13 contained HPV 6 and 11, 12 HPV 6, 7 HPV 11, and 1 HPV 6, 11, and 18. Two of four severe anal dysplasias contained HPV 16, whereas one case each of mild and moderate anal dysplasia contained HPV 6. No HPV DNA was detected in the anal squamous cell carcinomas. The study demonstrated the presence of HPV DNA in 73% of condylomata and 67% of anal dysplasias. The observations suggest that the cloacogenically derived anal epithelium is susceptible to infection by the same HPV types as infect the similarly derived epithelium of the lower female genital tract and that these HPV types result in some similar lesions, i.e., condylomata and dysplasias in both sites. A role in the genesis of anal cancer was not found in this study. PMID- 2546420 TI - Sensitivity and detection efficiency of the peroxidase antiperoxidase (PAP), avidin-biotin peroxidase complex (ABC), and peroxidase-labeled avidin-biotin (LAB) methods. AB - The authors have examined the sensitivity and detection efficiency of the three peroxidase methods that currently have the widest application in diagnostic immunohistochemistry: the peroxidase-antiperoxidase (PAP), the avidin-biotin complex (ABC), and the labeled avidin-biotin (LAB) methods. Sensitivity was evaluated by determining the highest useful dilution of polyclonal antiglucagon antibodies applied to formalin-fixed, paraffin-embedded human pancreas. Detection efficiency was evaluated by tabulation of the total number of positive (three or more positive cells) islets. On direct comparison, the LAB method exceeded the PAP and ABC methods in both sensitivity and detection efficiency, which were essentially equal. Titration of linking antiserum of the PAP method boosted its sensitivity and detection efficiency above that of ABC; the PAP had equal sensitivity to the LAB and exceeded it in detection efficiency. The authors conclude that comparisons of immunohistologic methods are meaningful only if both sensitivity and efficiency are considered along with the unique requirements of any single method. PMID- 2546421 TI - Cytomegalovirus infection involving the skin in immunocompromised hosts. A clinicopathologic study. AB - Cytomegalovirus (CMV) infection involving the skin in three transplant patients is presented. Patient 1, whose infection apparently was localized only to a cutaneous wound induced by extravasated ionotropic solution, survived. Mixed CMV and Candida infections developed in patient 2 in the cutaneous ulcer. He died of disseminated herpes simplex virus infection in two weeks. Patient 3 had CMV pneumonia and purpuric maculopapular eruption. He died of Pseudomonas sepsis 17 weeks later. Eighteen cases with CMV skin lesions are reported in the English literature. The clinical findings and the outcome of the current and the reported cases are analyzed. All patients were immunocompromised. CMV infection, when detected in the skin, appears to be associated with grave prognosis. Seventeen of 20 patients whose final outcome was recorded died within six months after the onset of CMV skin lesions. The outcome of one case is unknown. The mortality was 85%. The fatal cases had either concurrent disseminated CMV infection or mixed cutaneous or systemic infections. When the infection is localized in the skin wounds, the prognosis seems fairly good. All three such patients survived. PMID- 2546422 TI - Sexual activity, contraception, genital infections, and cervical cancer: support for a sexually transmitted disease hypothesis. AB - A case-control study was conducted in Utah between 1984 and 1987 to examine risk factors for cervical cancer. Interviews were completed with 266 histologically confirmed carcinoma in situ and invasive squamous cell cervical cancer cases who were categorically matched by age to 408 controls. Among the factors identified as altering risk for cervical cancer, after adjustment for age, education, church attendance, and cigarette smoking, were: having numerous sex partners (odds ratio (OR) = 8.99 for 10 or more partners); the current mate having several sex partners (adjusted OR for 10 or more partners = 8.62); using foam or jelly as a contraceptive method (OR, adjusted for number of sex partners, = 0.44); reported Trichomonas infection (OR, adjusted for number of sex partners, = 2.10); and herpes simplex virus type 2 infection as determined by 2:1 neutralization index values above 100 (OR = 2.70). A protective effect was noted from the use of diaphragms (OR = 0.67) or condoms (OR = 0.53) in women who reported more than one sex partner. These data support the hypothesis that cervical cancer is a sexually transmitted disease. PMID- 2546424 TI - Re: "Condyloma and Intraepithelial Neoplasia of the Uterine Cervix: a Case Control Study". PMID- 2546423 TI - Retroviral infections (HIV-1, HIV-2, and HTLV-I) in rural northwestern Tanzania. Clinical findings, epidemiology, and association with infections common in Africa. AB - During a three-week period in March/April 1987, the authors examined 253 consecutive patients referred to a rural hospital in northwestern Tanzania. Sera were tested for antibodies to human immunodeficiency virus type 1 (HIV-1), human immunodeficiency virus type 2 (HIV-2), and human T-lymphotropic virus type I (HTLV-I), as well as for various parasites, hepatitis B virus, and Treponema pallidum. Neopterin (urinary and serum) was chosen as the immunologic parameter. In eight of the 253 patients (3.2%), a clinical diagnosis of acquired immunodeficiency syndrome (AIDS) was established. Three of the AIDS patients had HIV-1 antibodies, two had HIV-1 antigen, one had both HIV-1 and HIV-2 antibodies, and in one patient, only HIV-2 antibodies were found. The total HIV-1 and HIV-2 seroprevalence (antibodies plus antigen) was 4.3%; HTLV-I seroprevalence was 9.9%. No correlation could be found between HIV (or HTLV-I) seropositivity and raised levels of antibody to the above pathogens. There was, however, a significantly positive correlation between HIV seropositivity and history of gonorrhea, whereas a history of operations, injections, vaccinations, blood transfusions, or scarification did not influence the level of HIV seropositivity. The most frequently noted epidemiologic association with HIV seropositivity was traveling to or coming from Uganda or Rwanda. Two thirds of the studied Tanzanians had elevated neopterin levels, and all seven HIV-seropositive patients with clinical signs of AIDS had extremely high serum and urinary neopterin levels compared with HIV-seropositive patients without signs of AIDS. Increased neopterin levels reflect a stimulation of the T-cell/macrophage system. PMID- 2546425 TI - Thyrotropin modulates receptors for atrial natriuretic peptide on intact human thyroid cells. AB - Interest in the mechanism of impaired salt and water metabolism in hypothyroidism has led to growing evidence of an interaction between atrial natriuretic peptide (ANP) and the thyroid, which includes reports of direct effects of thyroid hormone on ANP synthesis and circulating ANP levels, and of the presence of specific ANP receptors in human thyroid tissue, which may act to inhibit thyroglobulin (Tg) secretion. The authors questioned whether or not thyrotropin (TSH) has a role in this interaction. They used 125I-ANP to study the effect of TSH on ANP binding to human thyroid cells in primary culture. Binding competition by increasing concentrations of unlabeled ANP in the presence or absence of TSH was assessed by Scatchard analysis. At lower temperatures of 4 degrees C or 23 degrees C, TSH had no effect either on the ANP receptor equilibrium dissociation constant (Kd) or number of binding sites. However, at 37 degrees C, bovine TSH at 1 mU/ml reduced measurable binding sites by about 50% without affecting receptor affinity (Kd = 0.2 nM). Prolonged (6 days) coincubation of TSH with thyroid cells decreased the assayable ANP receptor. The effects of TSH appear to be specific because human luteinizing hormone, follicle-stimulatory hormone, growth hormone, human chorionic gonadotropin and iodide had no effect on ANP binding. Thus, human thyroid cells possess a single class of high-affinity, specific receptors for ANP with binding activity that is temperature dependent and modulated by TSH at physiologic temperature. TSH-mediated reduction of binding at 37 degrees C but not at 4 degrees C suggests an energy-dependent process that acts possibly by activating an ANP degradative enzyme or by changing the rate of receptor internalization and subsequent degradation. PMID- 2546426 TI - Trisomy 18 and hepatoblastoma. AB - A 4-month-old girl with trisomy 18 had a congenital heart defect and an hepatoblastoma. PMID- 2546427 TI - Gestational trophoblastic disease: a case-control study from the People's Republic of China. AB - A case-control study involving 331 patients with complete hydatidiform mole and 662 community controls matched to the cases on age and timing of pregnancy was conducted in Beijing, China. A history of a term birth was associated with reduced risk (odds ratio = 0.6, 95% confidence interval 0.4 to 0.9), with some evidence of further decrease with multiple births. Previous spontaneous abortions were not related to risk, although those with a prior induced abortion were at elevated risk, particularly if two or more abortions were involved (odds ratio = 2.8, 95% confidence interval 1.4 to 5.7). A history of having sought medical advice for infertility was associated with reduced risk (odds ratio = 0.5, 95% confidence interval 0.2 to 0.8), but those who reported use of herbal medicines during a first trimester of a previous pregnancy were at excess risk (odds ratio = 2.2, 95% confidence interval 1.3 to 3.6). In addition, a statistically significant trend in risk was observed with years of oral contraceptive use (odds ratio = 2.6, 95% confidence interval 0.9 to 6.9 for greater than or equal to 4 years of use). Dietary habits and family histories of cancer or trophoblastic disease were not related to risk in this study. PMID- 2546428 TI - Increased frequency of detection of human papillomavirus deoxyribonucleic acid in exfoliated cervical cells during pregnancy. AB - Exfoliated cells of the uterine cervix obtained from women during pregnancy and at the time of their first postpartum examination were used to monitor the prevalence of human papillomavirus infections in this population and to study the natural fluctuations in viral expression. When deoxyribonucleic acid hybridization analysis alone was used to monitor the presence of human papillomavirus infection, 20.9% of our study population had results that were positive for human papillomavirus deoxyribonucleic acid during their first trimester examinations. A dramatic increase in the percentage of women with positive results for human papillomavirus deoxyribonucleic acid was observed at the time of the patients' third-trimester examinations (46%). The overall increase in human papillomavirus-positive patients was a combination of a small number of patients who had positive results on their first examination and negative results on their second examination, and a larger number of patients who had negative results on their first-trimester examination and positive results for human papillomavirus deoxyribonucleic acid in the exfoliated cervical cells at the time of their third-trimester examination. The total percentage of patients with positive results for human papillomavirus deoxyribonucleic acid in their cervical cells at one or both assay points during pregnancy was 52.5%. Samples obtained at the postpartum examination demonstrated a dramatic decrease in the number of samples positive for human papillomavirus deoxyribonucleic acid (17.5%). This result was a combination of a large decrease in human papillomavirus-positive patients coupled with a small increase in detectable levels of human papillomavirus deoxyribonucleic acid in cervical samples from patients who had negative results on their previous examination. This study demonstrates a very high level of detectable human papillomavirus deoxyribonucleic acid in exfoliated cervical cells obtained during pregnancy and shows that the detectable levels of human papillomavirus deoxyribonucleic acid fluctuate during pregnancy. PMID- 2546429 TI - Mechanism for human papillomavirus transmission at birth. AB - We attempted to investigate mechanisms, in addition to sexual contact, by which human papillomaviruses associated with anogenital tract lesions could be transmitted. Samples of exfoliated cervical cells were obtained from 45 pregnant women and were assayed by Southern blot hybridization analysis for the presence of human papillomavirus nucleic acids. Twenty-five of the 45 women had cells positive for human papillomavirus deoxyribonucleic acid. A neonatal nasopharyngeal aspirate was obtained at term and analyzed for the presence of human papillomavirus deoxyribonucleic acid. We documented the presence of human papillomavirus deoxyribonucleic acid in the oral pharyngeal cavity of the neonates in 15 of 45 nasopharyngeal samples analyzed. Amniotic fluid was obtained from 13 patients when their membranes were artificially ruptured. These samples were assayed for the presence of human papillomavirus deoxyribonucleic acid; two of the 13 amniotic fluid samples contained human papillomavirus deoxyribonucleic acid. The detection of human papillomavirus deoxyribonucleic acid in the oral cavity of neonates is indicative of a perinatal mechanism of viral transmission. The detection of human papillomavirus deoxyribonucleic acid in the amniotic fluid may suggest an in utero mechanism of transmission. However, problems encountered in collecting the amniotic fluid samples preclude us from definitive interpretation of these data. PMID- 2546430 TI - Incus replacement prostheses of hydroxylapatite in middle ear reconstruction. AB - Hydroxylapatite is a calcium bioceramic that has the same chemical composition as living bone, Ca10 (PO4) 6 (OH) 2. Since 1970 it has been used as a material in reconstructive prostheses and augmentation of lost tissues in various surgical specialties including maxillofacial surgery, plastic surgery, otolaryngology, and orthopedics. For over 20 years the author has used autograft and homograft ossicles in tympanoplasty. These incudi have been modified into prostheses that were utilized in ossicular reconstruction. During this time two principle prostheses have evolved, the notched incus with short and long processes. The short process prosthesis is used with an intact stapes, whereas the notched incus with long process carries the sound pressure directly to the stapedial footplate. These prostheses have been successful in improving and maintaining hearing following tympanoplasty. Unfortunately, however, the use of human tissue has certain limitations: it is not readily accessible and has a limited shelf life. Furthermore, clinicians are wary of using homograft tissue as concern over the AIDS virus spreads. Therefore it was felt prudent to develop a manmade prosthesis that would as nearly as possible match the advantages of living bone. Hydroxylapetite most nearly met those qualifications. PMID- 2546431 TI - Biocompatible implants in tympanoplasty. AB - The history of implants in otology is reviewed. Currently available biocompatible ossicular implants are described, and a survey of their use by members of the American Otological Society and the American Neurotology Society is reported. PMID- 2546432 TI - cAMP delays disappearance of gap junctions between pairs of rat hepatocytes in primary culture. AB - Freshly isolated adult rat hepatocytes were found to be coupled through gap junctions, but coupling decreased abruptly 5-8 h after plating the cells on plastic culture dishes in physiological saline containing insulin and fetal calf serum. Loss of intercellular coupling was associated with disappearance of 27-kDa gap junction protein and of gap junctions seen by electron microscopy or immunocytochemistry. This disappearance of coupling was delayed approximately 8 h by treatment of the cultures with membrane permeant adenosine 3',5'-cyclic monophosphate (cAMP) [but not guanosine 3',5'-cyclic monophosphate (cGMP)] derivatives. Levels of gap junction protein and anatomically identified junctions were also maintained by 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP). Level of mRNA encoding the gap junction protein was maintained longer in cells treated with 8-BrcAMP than in untreated cells, but 8-BrcAMP did not detectably increase the transcription rate. Thus prolongation of coupling must result at least partially from extension of the lifetime of gap junction mRNA, allowing translation of message and assembly of channels for a longer period after plating. Treatment of cells with mRNA or protein synthesis inhibitors (alpha amanitin and cycloheximide) prolonged coupling to a similar extent as did treatment with 8-BrcAMP. alpha-Amanitin blocked transcription of gap junction mRNA, but levels of cytoplasmic mRNA encoding the 27-kDa gap junction protein were maintained, presumably by block of transcription of an mRNA degrading factor. The factor is probably a protein, since a similar effect on mRNA level was produced in cycloheximide-treated cells. Cells cultured in control medium were also observed to flatten as they became uncoupled, whereas cells cultured for as long as 16 h in elevated 8-BrcAMP remained round and well coupled. The correlation between shape and coupling strength was not obtained after treatment with the microtubule stabilizing agent, taxol, which maintained the spherical shape of the cells but did not delay the disappearance of dye coupling. Nocodazole, which blocks microtubule formation, also maintained the spherical shape of the cells but delayed the disappearance of dye coupling. In addition to gating by covalent modification or other mechanisms, hormones and drugs that alter the intracellular cAMP concentration may affect intercellular communication by changing the lifetime of the mRNA encoding the main gap junction protein, thereby decreasing or increasing its synthesis. In addition, cAMP may act by decreasing removal of junctions from appositional membranes. PMID- 2546433 TI - Regulation of Na+ and K+ contents in rat thymocytes. AB - A modified nystatin technique allowed the investigation of the initial rate of Na+ efflux as a function of internal Na+ content under steady-state conditions in rat thymocytes. This kinetic study showed that 1) ouabain-sensitive Na+ efflux as a function of internal Na+ can be adjusted by a three-sites kinetic model, with a maximal pump rate of 581 +/- 79 mmol.l cells-1.h-1 and an apparent dissociation constant for internal Na+ of 10.0 +/- 1.0 mmol/l cells (mean +/- SE of 3 experiments), 2) bumetanide-sensitive Na+ efflux was extremely low compared with the pump efflux (approximately 1%), and 3) ouabain- and bumetanide-resistant Na+ efflux was almost a linear function of internal Na+ content (as expected for a Na+ leak). This "all-pump" mechanism of thymocyte Na+ regulation was confirmed by non-steady-state experiments showing that 1) ouabain induced a rapid net Na+ gain and K+ depletion in fresh thymocytes and completely blocked the recovery of normal cation contents in Na+-loaded-K+-depleted thymocytes, and 2) bumetanide was unable to modify thymocyte Na+ and K+ contents. Na+ extrusion by Na+-loaded thymocytes was unaffected by prostaglandin E2, isoproterenol, or platelet aggregating factor (PAF) and was slightly impaired in the adult spontaneously hypertensive rat of the Okamoto strain (10% lower rate constant for net Na+ extrusion, P less than 0.05). Concerning cell Na+ regulation, our results do not support the concept that rat thymocytes are more representative of vascular cells than enucleated erythrocytes. PMID- 2546434 TI - Regulation of Na-K-Cl cotransport in endothelial cells by atrial natriuretic factor. AB - Many vasoactive agents have been shown to bind to specific receptors on endothelial cells. Among these is atrial natriuretic factor (ANF). Binding of ANF to endothelial cells has been demonstrated to induce elevation of intracellular guanosine 3',5'-cyclic monophosphate (cGMP). Other vasoactive agents have been shown to cause elevation of intracellular adenosine 3',5'-cyclic monophosphate (cAMP), Ca, and diacylglycerol. However, the endothelial cell response that occurs subsequent to elevation of cGMP or other second messengers is not well understood. Recently, endothelial cells have been shown to possess a Na-K-Cl cotransport system that is stimulated by vasopressin and bradykinin and inhibited by isoproterenol. Thus it is possible that modulation of Na-K-Cl cotransport may play a role in the endothelial cell response to second messengers that are elevated by ANF and other vasoactive agents. This possibility was examined in the present study by evaluating the effects of a variety of vasoactive agents and their second messengers on endothelial cell Na-K-Cl cotransport. Cotransport was assessed as bumetanide-sensitive K influx in cultured bovine aortic endothelial cells. A number of agents were found to reduce Na-K-Cl cotransport, including ANF, acetylcholine, histamine, and norepinephrine. Cotransport was found to be stimulated by angiotensin II, as well as vasopressin and bradykinin. Na-K-Cl cotransport was also inhibited by elevation of intracellular cGMP or cAMP or by treatment of the cells with phorbol ester to activate protein kinase C. However, A23187-induced elevation of intracellular Ca caused stimulation of Na-K-Cl cotransport.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546435 TI - Regulation of ion transport in porcine gallbladder: effects of VIP and norepinephrine. AB - The objective of this study was to investigate the effects of vasoactive intestinal peptide (VIP) and norepinephrine (NE) on Na and Cl transport across the isolated porcine gallbladder. Serosal addition of either VIP or secretin increased the short-circuit current (Isc). The half-maximal effect for VIP was 84.3 nM. The effect of VIP was mimicked by 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP). Replacement of Cl with gluconate nearly abolished the effect of 8-BrcAMP on Isc, whereas HCO3 replacement with N-2 hydroxyethylpiperazine-N'-2-ethane-sulfonic acid buffer had no effect. Transepithelial flux measurements indicated that 8-BrcAMP stimulates net Cl secretion and inhibits Na absorption. Norepinephrine inhibits VIP-stimulated changes in Isc as well as the basal Isc. NE does not, however, reverse the effects of 8-BrcAMP on Isc. The effects of NE are antagonized by yohimbine (alpha 2-adrenergic receptor antagonist) but not prazosin (an alpha 1-adrenergic receptor antagonist). VIP causes a 2.5-fold increase in cAMP content in the gallbladder epithelium. This increase is blocked by NE. Serosal tetrodotoxin did not inhibit the peptide effects, indicating that VIP receptors are localized on the epithelium. Depolarization of submucosal nerves with veratrine inhibited the basal Isc and was reversible with yohimbine. This result indicated that sympathetic nerve pathways regulate Na and Cl absorption in vitro. PMID- 2546436 TI - Atrial natriuretic peptide and nitroprusside cause relaxation of cultured rat mesangial cells. AB - These studies evaluate the effects of atrial natriuretic peptide (ANP) and nitroprusside (NP) on cultured mesangial cells (MC) grown on a flexible silicone rubber surface. The basal tone of the MC produced wrinkles on the silicone rubber surface. A decrease in number or magnitude of wrinkles was considered to represent cell relaxation, whereas an increase represented cell contraction. ANP (10(-9) M) produced a relaxation in greater than 60% of the cells by 10 min. The percentage of cells showing a decrease of wrinkles was significantly higher (P less than 0.05 at 5 min and P less than 0.001 at 10 min) during the ANP-treated period than during the control period. NP (10(-5) M) caused a decrease of wrinkles in greater than 80% of cells (P less than 0.02 at 5 min and P less than 0.01 at 10 min) compared with a 5% decrease in the control period. Dibutyryl guanosine 3',5'-cyclic monophosphate (DBcGMP; 10(-4) M) also produced a decrease of wrinkles in 81% of the cells (P less than 0.02) compared with a 9% decrease in the control period. MC treated with ANP, NP, or DBcGMP and then labeled with 7 nitrobenz-2-oxa-1,3-diazol-4-yl (NBD)-phallacidin did not show obvious alteration of morphology of actin filaments compared with untreated cells. ANP could inhibit as well as partially reverse the agonist (angiotensin II)-induced contractile response. ANP (10(-10)-10(-8) M) as well as NP (10(-5) M) increased intracellular cGMP content of MC (P less than 0.005) compared with control cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546437 TI - Receptors for IGF-I, but not for IGF-II, on proximal colon epithelial cell apical membranes. AB - Rabbit proximal colon epithelial cell apical membranes, which are known to contain receptors for insulin, were isolated by a Ca2+-precipitation technique. Binding assays with 125I-insulin-like growth factor I (IGF-I) revealed the presence of specific high-affinity binding sites, with 50% inhibition of binding observed at a concentration of 13.7 ng/ml IGF-I. In contrast, 50% inhibition of 125I-IGF-I binding was observed at an insulin concentration of 1.37 micrograms/ml, suggesting that 125I-IGF-I was not binding to insulin receptors present in this tissue. Cross-linking studies revealed an 125I-IGF-I binding subunit of relative molecular weight (Mr) of 130,000 under reducing conditions on docecyl sulfate-polyacrylamide gel electrophoresis that was similar to the IGF-I binding subunit in human placental membranes (Mr 140,000). Binding and cross linking studies with 125I-insulin-like growth factor II (IGF-II), however, failed to reveal a specific receptor for this peptide in colon epithelial cell membranes. These results establish the coexistence of receptors for IGF-I and insulin, but not IGF-II, on rabbit proximal colon epithelial cell apical membranes and demonstrate that colon epithelial cells are capable of selective synthesis of various peptide hormone receptors. PMID- 2546438 TI - Effect of short- and long-term beta-adrenergic blockade on lipolysis during fasting in humans. AB - Stable isotope tracers and indirect calorimetry were used to evaluate the importance of beta-adrenergic stimulation of lipolysis and triglyceride-fatty acid cycling during fasting in healthy human volunteers. Each subject was studied after 12 and 84 h of fasting both with and without propranolol infusion (protocol 1) and when oral propranolol treatment was given throughout fasting (protocol 2). In protocol 1, the rates of appearance of glycerol and palmitic acid increased from 3.04 +/- 0.19 and 1.78 +/- 0.17 mumol.kg lean body mass-1.min-1, respectively, after 12 h of fasting to 5.28 +/- 0.31 and 3.47 +/- 0.15 mumol.kg lean body mass-1.min-1, respectively, after 84 h of fasting (P less than 0.005). The rate of triglyceride-fatty acid cycling increased from 97 +/- 8 to 169 +/- 5 mumol/min (P less than 0.005). Intravenous propranolol infusion decreased the rate of lipolysis after both 12 and 84 h of fasting, but the magnitude of the antilipolytic effect was much greater after 84 h (P less than 0.005). In protocol 2, the rate of lipolysis and triglyceride-fatty acid cycling was still increased by fasting despite beta-adrenergic blockade with oral propranolol. This study demonstrates that beta-adrenergic stimulation contributes to the mobilization of fat during fasting. However, other mechanism(s) can increase lipolysis and triglyceride-fatty acid cycling when beta-adrenergic receptors are continuously blocked. PMID- 2546439 TI - Hormonal and metabolic responses to exercise in humans: effect of sensory nervous blockade. AB - Previous studies have indicated that motor center ("feedforward") activity is important for hormonal and metabolic responses to exercise. Now, epidural blockade at vertebrae L3-L4 was used to evaluate the importance of afferent neural feedback from working muscles. Six healthy, young males cycled for 20 min at 55 +/- 4% (mean +/- SE) of maximal oxygen uptake with, as well as without, epidural anesthesia. During anesthesia cutaneous sensory blockade was present below segment T11-12, the postexercise ischemic pressor response was attenuated from 34 +/- 9 to 14 +/- 4 mmHg, muscle strength reduced to 80 +/- 5% of control, and perceived exertion (Borg scale) was increased. At rest hormonal and metabolic parameters did not change in response to epidural anesthesia. During exercise, responses of catecholamines, insulin, glucagon, and growth hormone (GH) in plasma as well as glucose production and utilization, plasma free fatty acids, and plasma glycerol were similar in epidural and control experiments (P greater than 0.05). In contrast during submaximal exercise, plasma concentrations of adrenocorticotropin (ACTH) and beta-endorphin increased only in experiments without epidural anesthesia. The data indicate that impulses in afferent nerves from the working muscles are essential for the ACTH and beta-endorphin responses to submaximal dynamic exercise in humans. Afferent nervous activity is probably less important than efferent activity from motor centers for responses of GH, catecholamines and insulin, and, in turn, extramuscular fuel mobilization in exercise. PMID- 2546440 TI - Intestinal alkaline phosphatase is secreted bidirectionally from villous enterocytes. AB - A fraction of intestinal alkaline phosphatase (IAP) is secreted into blood. To study this process, enzyme secretion was examined in a fetal (IRD-98) and a differentiated (Caco-2) intestinal cell line. Tissue-unspecific alkaline phosphatase (AP) activity in the IRD-98 cells increased 20-fold after addition of 1.5 mM sodium butyrate and 40 mM NaCl, but no AP activity was secreted into the medium. In contrast, newly synthesized IAP in Caco-2 cells was secreted into the medium. AP secretion increased with time and was inhibited by monensin. Medium AP was still partially bound to membranes as assessed by Triton X-114 phase separation and could be released by the addition of serum. Analysis by sodium dodecyl sulfate polyacrylamide gels and by isoelectric focussing showed that secreted AP gave a pattern similar to that of the AP released from membranes by phospholipase D treatment. When Caco-2 cells were grown on filters, AP activity was found in both basolateral (75%) and luminal (25%) media. These data demonstrate that the secretion of a particulate AP with extracellular release from the membrane can account for the appearance of the intestinal isozyme in both the serum and the lumen. PMID- 2546441 TI - Inhibition of stimulated lacrimal secretion by [D-Ala2]Met-enkephalinamide. AB - The synthetic enkephalin analogue, [D-Ala2]Met-enkephalinamide (DALA) was used to investigate opioid peptide modulation of lacrimal protein secretion. By use of an in vitro perifusion system, the secretion of peroxidase by rat lacrimal gland fragments was measured during continuous stimulation for up to 60 min. DALA had no effect on unstimulated secretion of peroxidase. However, the addition of DALA to the perifusion medium resulted in a dose-dependent (10 nM-10 microM) inhibition of carbachol-stimulated peroxidase release. The maximum effect of DALA was achieved at a dose of 10 microM, which resulted in a 54% inhibition of carbachol-induced secretion. The opiate antagonist naloxone (10 nM-10 microM) did not alter basal or carbachol-stimulated peroxidase release. The effect on 10 microM carbachol-stimulated secretion by the addition of 3 microM DALA, however, was reversed in a dose-dependent manner by naloxone. The extent of inhibition of vasoactive intestinal peptidergic (VIPergic) stimulation (50 nM VIP) by DALA was similar to the inhibition of cholinergic stimulation of peroxidase release by gland fragments. Neither alpha 1-adrenergic stimulation of secretion nor a synergistic stimulation by VIP and carbachol was inhibited by the enkephalin analogue. We conclude that DALA exerts an inhibitory modulation of cholinergic and VIPergic stimulation of in vitro lacrimal protein secretion and suggest a physiological role for methionine enkephalin as an inhibitory peptide involved in the regulation of lacrimal gland function. PMID- 2546442 TI - Regulation of plasma motilin by opioids in the dog. AB - In the first part of this study, we compared the effects of morphine and trimebutine, two opioid receptor agonists, on small intestinal motility and plasma motilin in dogs. Morphine (100 micrograms/kg iv for 10 min) induced first a typical vomiting myoelectric profile followed subsequently by a migrating electrical activity mimicking phase III of the migrating myoelectric complex; trimebutine (5 mg/kg iv for 10 min) initiated only a migrating phase III-like activity. Despite their different initial contractile effects, both agents induced a significant and similar rise in plasma motilin that preceded the beginning of the premature phase III. In the second portion of the study, naloxone, an opioid receptor antagonist, was infused to verify the influence of endogenous opiates on plasma motilin and on the migrating motor complex. Naloxone (2 mg/kg, then 0.5 mg.kg-1.h-1 iv) delayed significantly the cyclic recurrence of plasma motilin peak increases and of the phase IIIs. In some animals, where naloxone abolished the phase IIIs, the amplitude of the motilin peak increases was significantly diminished. These results suggest 1) that opioid administration increases plasma levels of motilin by a mechanism that is independent of the intestinal contractile activity, and 2) that endogenous opioids could be physiological inducers of plasma motilin increases in the conscious dog. PMID- 2546443 TI - Regulation of renal ion transport and cell growth by sodium. AB - Intracellular sodium has been implicated in a variety of cellular processes including regulation of Na+-K+-ATPase activity, mitogen-induced cell growth, and proliferation and stimulation of Na+-K+-ATPase by aldosterone. In renal epithelial cells a rise in sodium uptake across the apical membrane increases intracellular sodium concentration, which in turn stimulates the turnover rate of Na+-K+-ATPase and thereby enhances sodium efflux across the basolateral membrane. A prolonged increase in sodium uptake causes dramatic hypertrophy and hyperplasia and a rise in the quantity of Na+-K+-ATPase in the basolateral membrane. These structural and functional changes occur in the kidney in the absence of alterations in plasma aldosterone and vasopressin levels. Several mitogens induce growth and proliferation by initiating a cascade of events, which include a rise in intracellular sodium. Accordingly, an increase in the sodium concentration within renal epithelial cells may elicit a "mitogen-like" effect by initiating the cascade at the sodium step, even in the absence of a mitogen. A rise in cell sodium may also stimulate the production of autocrine growth factors that directly or indirectly regulate cell growth and proliferation, by modifying the response to mitogens or to changes in the ionic composition of the extracellular fluid. In this review we will examine the evidence that supports a role for intracellular sodium in regulating these cellular events in renal epithelial cells. PMID- 2546444 TI - K+ and Rb+ transport by the rabbit CCD: Rb+ reduces K+ conductance and Na+ transport. AB - We compared transport of K+ and Rb+ across the rabbit cortical collecting duct to gain insight into the mechanisms of K+ secretion. Passive tracer fluxes, active secretory rates, electrophysiological behavior, and the ability of each ion to support Na+-K+-ATPase activity were determined. When active transport was inhibited by amiloride, K+ permeability was twice the Rb+ permeability. Transepithelial conductance (GT) was half as great in solutions where 5 mM Rb+ replaced 5 mM K+. When 4 mM Ba2+ was added to the lumen, both Rb+ and K+ permeability fell to values not different from that expected for paracellular diffusion. The relationship between Ba2+-induced changes in the K+ and Rb+ permeabilities and in the simultaneously measured GT provides strong evidence that K+ transport across the apical membrane is largely, if not exclusively, conductive. We also determined that net K+ secretion is greater than net Rb+ secretion (when each is the abundant ion). The reasons for this difference probably involve several steps in the K+ secretory process and include the following: 1) reduced ATPase activity in the presence of Rb+ (approximately 80%) compared with K+, 2) reduction of Na+ absorption, and 3) partial blockade of the apical (and perhaps basolateral) K+ conductance. Although there were quantitative differences between K+ and Rb+ transport, we found no evidence suggesting that these ions are transported by different mechanisms. PMID- 2546445 TI - Inhibition of rat mesangial cell mitogenesis by nitric oxide-generating vasodilators. AB - Recent studies indicate that endothelium-derived relaxing factor (EDRF) may be identical with nitric oxide (NO). The purpose of this study was to investigate the antimitogenic effect of NO-generating drugs in cultured mesangial cells. S nitroso-N-acetylpenicillamine, sodium nitroprusside, and isosorbide dinitrate, which generate NO, dose dependently inhibited serum-stimulated DNA synthesis. All three drugs also inhibited the rate of cell proliferation, whereas sodium nitroprusside and S-nitroso-N-acetylpenicillamine decreased cell density at confluence. The antimitogenic activity of S-nitroso-N-acetylpenicillamine was labile in culture medium and could be inhibited by hemoglobin, supporting the view that NO, in free or bound form, was the ultimate effector. All three vasodilators increased cellular guanosine 3',5'-cyclic monophosphate (cGMP) levels dose dependently; moreover, 8-bromo-cGMP mimicked the effects of the NO generating drugs, suggesting that cGMP may be an intracellular mediator of antimitogenesis. The growth-inhibitory effect of S-nitroso-N-acetylpenicillamine was reversible and was not due to cell toxicity as shown by several criteria of cell viability. The results raise the possibility that EDRF/NO may be a modulator of mesangial cell growth in vivo. PMID- 2546446 TI - Effects of Cl- transport inhibitors on Cl- permeability across hamster ascending thin limb. AB - The highly conductive Cl- transport pathway exists in the ascending thin limb (ATL) of Henle's loop. To characterize the mechanism of the Cl- conductance across the ATL, we examined effects on Cl- permeability across hamster ATL of Cl- transport inhibitors, including 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB), diphenylamine carboxylate (DPC), and anthracene-9-carboxylic acid (9-AC), by the in vitro microperfusion technique. NPPB added to the bath caused reversible suppression of the relative permeability of Cl- to Na+ (PCl/PNa), as estimated from the NaCl diffusion voltage in a dose-dependent manner in a range from 3 x 10(-6) to 10(-3) M. The concentration of NPPB that inhibited PCl/PNa by 50% (ID50) was approximately 3 X 10(-5) M. When 3 X 10(-5) M NPPB was added to the bath, the lumen-to-bath flux coefficient for 36Cl (Kl----b,Cl- 10(-7) cm2/s) was decreased from 130.7 +/- 7.3 to 52.2 +/- 11.6 (n = 7, P less than 0.01). Application of NPPB in the lumen also caused reversible suppression of PCl/PNa, but this effect was less potent compared with the application of the drug via the bath. Whereas 10(-3) M 9-AC or 10(-3) M DPC decreased PCl/PNa by 6.5 +/- 1.2 and 10.2 +/- 2.6%, respectively, 3 X 10(-4) M NPPB decreased PCl/PNa by 69.7 +/- 3.8%. In maleimide-treated tubules, addition of 10(-3) M N-ethylmaleimide (NEM) increased PCl/PNa from 1.0 +/- 0.1 to 1.8 +/- 0.1 (n = 7, P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546448 TI - Distribution of alpha 1-adrenergic receptors in myocytic regions and vasculature of feline myocardium. AB - The physiological and pathophysiological roles of myocardial alpha 1-adrenergic receptors are not clearly defined. To delineate the distribution of alpha 1 receptors in myocytic and vascular components of the heart, we characterized the binding of [3H]prazosin to alpha 1-receptors in unfixed, transmural slices of feline left ventricle. Specific binding ratios greater than 95% were achieved at radioligand concentrations near the dissociation constant (Kd). Binding of radioligand to receptors in transmural slices was rapid, reversible, saturable, stereoselective, and displaceable by subtype-selective antagonists with a rank order of potency characteristic of alpha 1-receptors. Analysis of binding isotherms indicated a Bmax of 9.1 +/- 1.9 fmol/mg protein and a Kd of 36.9 +/- 6.3 pM. Results of light microscopic autoradiography indicated that regions composed of closely arranged cardiac myocytes contained three to fourfold more alpha 1-receptors per unit section area than the resistance microvasculature, which in turn contained approximately twice the density of alpha 1-receptors observed in the medial smooth muscle of large conductance arteries. No differences in the density of alpha 1-receptors between subepicardial and subendocardial regions were observed for either myocytic regions or coronary arterioles. These results indicate that myocytic regions of the cat ventricle contain a high density of alpha 1-adrenergic receptors. The methods developed should be of value in characterizing the distribution and function of alpha 1 receptors and mechanisms of regulation of adrenergic responsiveness in intact myocyardium. PMID- 2546447 TI - Alpha 2-adrenoceptor stimulation can augment coronary vasodilation maximally induced by adenosine in dogs. AB - To determine whether alpha 2-adrenoceptor stimulation can augment adenosine induced coronary vasodilation, 34 open-chest dogs were studied. When a small dose of clonidine (up to 0.24 micrograms.kg-1.min-1 ic) was administered under beta adrenoceptor blockade, coronary blood flow [312 +/- 16 (SE) ml.100 g-1.min-1] maximally induced by intracoronary infusion of adenosine was further increased (P less than 0.05) by 66 +/- 16 ml.100 g-1.min-1, despite no significant changes in coronary perfusion pressure, myocardial oxygen consumption, and coronary venous adenosine concentration. However, when a larger dose of clonidine (0.36-0.60 micrograms.kg-1.min-1) was infused, adenosine-induced flow progressively decreased. This biphasic action of the alpha 2-adrenoceptor activity was also observed when the dose of norepinephrine was increased during alpha 1 adrenoceptor blockade with prazosin. Norepinephrine up to 0.24 micrograms.kg 1.min-1 (ic) further increased adenosine-induced coronary blood flow by 24 +/- 5% (P less than 0.001), whereas hyperemic flow was decreased by a larger dose of norepinephrine. In contrast to the alpha 2-adrenoceptor stimulation, the alpha 1 adrenoceptor stimulation (norepinephrine with yohimbine) progressively decreased coronary blood flow. Furthermore, with a small dose of clonidine, reactive hyperemic flow significantly increased compared with that without clonidine (303 +/- 13 vs. 355 +/- 13 ml.100 g-1.min-1, P less than 0.001), but a larger dose of clonidine adversely reduced reactive flow (254 +/- 18 ml.100 g-1.min-1, P less than 0.001). Adenosine release during reactive hyperemia with and without intracoronary infusions of clonidine were not altered significantly.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546449 TI - Effects of PAF and BN 52021 on cardiac function and regional blood flow in conscious rats. AB - The effect of intravenous injections (0.1-3 nmol/kg) of platelet-activating factor (PAF) on blood pressure, heart rate, cardiac output, and blood flow (hindquarter, renal, mesenteric) were studied in conscious rats. PAF decreased blood pressure and total peripheral resistance (TPR) but increased heart rate; cardiac output was reduced by the highest dose. Low doses of PAF increased blood flow and decreased vascular resistance in all vascular beds, whereas high doses reduced mesenteric blood flow in part by increasing mesenteric vascular resistance. The hypotensive and cardiac effects of PAF were blocked by intravenous infusions of the selective PAF-receptor antagonists, 15 mg/kg BN 52021 and 1 mg/kg SDZ 63-441. BN 52021 also attenuated the hindquarter and renal responses to PAF, but the mesenteric responses remained relatively unchanged. The results indicate that PAF is a potent vasodilator of mesenteric greater than hindquarter = renal vessels at low doses and a cardiac depressant at high doses. A therapeutic role for the PAF antagonists BN 52021 and SDZ 63-441 is suggested in endotoxemia, anaphylaxis, and other disease states in which increased release of PAF contributes to key hemodynamic derangements. PMID- 2546450 TI - Superoxide anion is an endothelium-derived contracting factor. AB - The calcium ionophore A23187 causes endothelium-dependent contractions in canine basilar arteries. Removal of the endothelium, or treatment with indomethacin or superoxide dismutase (SOD), prevented the endothelium-dependent excitatory effect of the calcium ionophore. Catalase and deferoxamine were without effect. Superoxide anion generated by xanthine plus xanthine oxidase in the presence of catalase caused contractions of the vascular smooth muscle, which were abolished by SOD or heat inactivation of xanthine oxidase. The A23187-induced production of prostaglandins F2 alpha and E2 and thromboxane B2 was abolished by the removal of endothelium and by treatment with indomethacin but was not affected by the presence of SOD plus catalase. These observations are consistent with the hypothesis that superoxide anion, rather than prostaglandins generated by hydroperoxidase activity of cyclooxygenase, is an endothelium-derived contracting factor in canine cerebral arteries. PMID- 2546452 TI - Potentiated cortisol response to paired hemorrhage: role of angiotensin and vasopressin. AB - Potentiated adrenocorticotropin (ACTH) and cortisol responses occur after the second of two small hemorrhages (hems) spaced 24 h apart in the dog. To test whether increased responses of other hormones might be associated with this effect, we examined plasma renin activity (PRA), angiotensin II (ANG II), and vasopressin after paired 10% hem (H1 and H2) spaced 5 h apart in chronically prepared conscious dogs. Cortisol secretion increased after each hem, and the response to H2 was larger (P less than 0.05; H1 peak at 6.8 +/- 1.3 micrograms/min vs. H2 peak at 18.3 +/- 5.3 micrograms/min). ACTH did not change after H1 but increased after H2, and the H2 response was larger (P less than 0.01). Vasopressin increased after each hem, and the H2 response was larger (P less than 0.01). The time courses of ACTH and vasopressin responses were similar after H2 (significant increases by 8 min). PRA and ANG II increased by 4 min after each hem, and although the difference was small the early PRA and ANG II responses were greater after H2. Blood volume and hem volume did not differ between hems. Hemodynamic responses to the hems were not different. We conclude that, although the PRA and ANG II respond rapidly enough after hem to influence pituitary responses, the slightly greater responses of these factors to H2 are not responsible for greatly increased pituitary-adrenal responses to H2. On the other hand, the markedly potentiated vasopressin response to H2, which parallels that of ACTH, suggests that vasopressin may mediate the increased ACTH responses to H2. PMID- 2546451 TI - Is there a second external lidocaine binding site on mammalian cardiac cells? AB - Lidocaine and its permanently charged analogue QX-314 block sodium current (INa) in nerve, and by this mechanism, lidocaine produces local anesthesia. When administered clinically, lidocaine prevents cardiac arrhythmias. Nerve and skeletal muscle are much more sensitive to local anesthetics when the drugs are applied inside the cell, indicating that the binding site for local anesthetics is located on the inside of those Na channels. Using a large suction pipette for voltage clamp and internal perfusion of single cardiac Purkinje cells, we demonstrate that a charged lidocaine analogue blocks INa not only when applied from the inside but also from the outside, unlike noncardiac tissue. This functional difference in heart predicts that a second local anesthetic binding site exists outside or near the outside of cardiac Na channels and emphasizes that the cardiac Na channel is different from that in nerve. PMID- 2546453 TI - Effect of upper airway CO2 pattern on ventilatory frequency in tegu lizards. AB - Nasal CO2-sensitive receptors are reported to depress ventilatory frequency in several reptilian species in response to constant low levels of inspired CO2. The purpose of this study was to determine the influence of phasic patterns of CO2 in the upper airways on ventilation. Awake lizards (Tupinambis nigropunctatus) breathed through an endotracheal tube from an isolated gas source. A second gas mixture was forced at constant flow into the external nares. A concentration of 4% CO2 was intermittently pulsed through the nares in a square-wave pattern with a frequency of 60, 12, 6, 4.2, 1.8, and 0.6 cycles/min. Concentrations of 2, 3, 4, and 6% CO2 were also pulsed through the nares at 12 cycles/min and compared with sustained levels of 1, 1.5, 2, and 3%. Additionally, 0 or 3% CO2 was forced through the upper airways with a servo system designed to mimic normal ventilatory flow and gas concentrations. No changes in breathing pattern were noted during any of the pulsing protocols, although a significant breathing frequency depression was present with sustained levels of CO2 of comparable mean concentrations. We conclude that ventilatory control is selectively responsive to sustained levels of environmental CO2 but not to phasic changes in upper airway CO2 concentration. PMID- 2546454 TI - Lack of vasopressin increases hypothalamic atrial natriuretic peptide binding sites. AB - Atrial natriuretic peptide (ANP) binding sites were measured by quantitative autoradiography in the supraoptic and paraventricular nuclei and in the subfornical organ of hypophysectomized, adrenalectomized, and genetically vasopressin-deficient (Brattleboro) rats. Hypophysectomized and Brattleboro rats had significantly higher numbers of ANP binding sites in the supraoptic nucleus and in the magnocellular subdivision of the paraventricular nucleus than their respective controls. ANP binding density was also increased in the parvocellular subdivision of the paraventricular nucleus in hypophysectomized rats and in the subfornical organ of homozygous Brattleboro rats. When homozygous Brattleboro rats were treated with vasopressin, the density of ANP binding sites was restored to control level in the subfornical organ but not in the supraoptic or paraventricular nuclei. Adrenalectomy did not influence ANP binding in the brain areas studied. Increased ANP binding density in Brattleboro rats and after hypophysectomy in the nuclei in which vasopressin neurons are located suggest that ANP binding sites may represent physiologically active receptors and may mediate the inhibitory action of ANP on vasopressin secretion. PMID- 2546455 TI - Acute-phase response to endogenous pyrogen in rabbit: effects of age and route of administration. AB - Aspects of host defense, collectively called the acute phase response (APR), can be induced by central actions of cytokines. To determine whether aging alters this response, aged and young rabbits were given endogenous pyrogen (EP), a crude preparation containing interleukin 1 and other cytokines, via an intracerebroventricular cannula. Arterial blood was sampled before EP administration and 2, 4, and 24 h later. Measurements were made of changes in body temperature, white blood cells, neutrophils, concentration of antipyretic, anti-inflammatory peptide alpha-melanocyte stimulating hormone (alpha-MSH), corticosterone, and C-reactive protein (CRP) concentrations. EP caused greater fever and increases in corticosterone and CRP in young rabbits. EP-induced changes in circulating neutrophils did not show age-related differences. There was no significant change in alpha-MSH in either age group; thus only certain aspects of APR induced by central actions of EP were altered with aging. To determine whether changes in APR caused by peripherally administered cytokines are similar to those after central injection, young female rabbits were given EP by both routes. Although administration caused greater fever, increases in alpha MSH and corticosterone concentrations and in neutrophil counts were greater after intravenous administration, perhaps the result of a combined influence on peripheral and central receptors. The EP-induced increase in circulating alpha MSH is a new finding that indicates that this antipyretic and anti-inflammatory peptide is rapidly available to modulate host responses after challenge. PMID- 2546456 TI - Mechanism of inhibition of renin response to hypotension by atrial natriuretic factor. AB - We have reported that infusion of atrial natriuretic factor (ANF) inhibited the rise in plasma renin activity (PRA) in response to constriction of the abdominal aorta to cause a reduction in renal perfusion pressure (RPP). To evaluate the effect of ANF on neural control of renin release, acute thoracic inferior vena caval constriction (TIVCC) was performed in conscious dogs to reduce arterial pressure by 25% of control and stimulate PRA by a reflex increase in renal nerve activity and a reduction in RPP. Propranolol was used to block neural stimulation of renin release. TIVCC caused significant increases in PRA, plasma aldosterone, arginine vasopressin (AVP), and adrenocorticotropic hormone (ACTH) concentrations. The increase in PRA was significantly reduced by the infusion of either ANF at 20 ng.kg-1.min-1 or propranolol. The combined infusion of ANF and propranolol produced an additive and complete inhibition of the renin response to TIVCC; therefore the effect of ANF is independent of neural stimulation of renin release. ANF at 20 ng.kg-1.min-1 also inhibited increases in aldosterone, AVP, and ACTH, but ANF at 5 ng.kg-1.min-1 only affected the aldosterone response to TIVCC. Therefore ANF inhibits angiotensin II-stimulated aldosterone synthesis and/or secretion at very low doses and at higher doses attenuates reflex increases in AVP and ACTH caused by hypotension. PMID- 2546457 TI - Site of action of putative lipostatic factor: hypothalamic metabolism of parabiotic rats. AB - Studies were conducted to determine whether metabolic adaptation occurred in the hypothalamus of overfed parabiotic rats and their partners to distinguish between the adaptations caused by increased caloric intake and those caused by the production of a "lipostatic factor." The induction of overfed obesity in one parabiotic partner was employed to test the hypothesis that a putative lipostatic factor produced in the obese parabiotic elicited the hypophagic-lipid-mobilizing effect observed in the lean parabiotic via alterations in hypothalamic fatty acid and glucose metabolism. Fatty acid oxidation in the ventrolateral hypothalamus (VLH) of overfed parabiotic rats and their partners was lower than in ad libitum parabiotic rats. Net flux of glucose through the VLH gamma-aminobutyric acid (GABA) shunt was elevated in overfed parabiotic rats compared with the net flux observed in their partners and ad libitum parabiotic rats, the levels being similar in these last two groups. Net flux of glucose through the ventromedial hypothalamic (VMH) pentose shunt in overfed parabiotic rats and their partners was elevated relative to ad libitum parabiotic rats. The putative lipostatic factor may act to regulate energy balance through modification of VLH fatty acid oxidation and/or glucose flux via the VMH pentose shunt. PMID- 2546458 TI - Analysis of lymphoepithelioma and lymphoepithelioma-like carcinomas for Epstein Barr viral genomes by in situ hybridization. AB - Lymphoepithelioma of the nasopharynx has a strong association with Epstein-Barr virus (EBV). To test the hypothesis that lymphoepithelioma-like carcinomas occurring at other sites are also associated with EBV virus, we used in situ hybridization to analyze 20 cases of lymphoepithelioma and histologically similar lesions and five basaloid squamous cell carcinomas for evidence of EBV genomes. EBV genomes were demonstrated in six of six lymphoepitheliomas of the nasopharynx but in none of five basaloid squamous cell carcinoma. Only one of 14 lymphoepithelioma-like carcinomas was found to contain EBV genomes. The single positive lymphoepithelioma-like carcinoma occurred in the lung of an Asian patient, suggesting that ethnic or geographic influences may be important in determining whether EBV is associated with these nonnasopharyngeal neoplasms. Despite their histologic similarity, most lymphoepithelioma-like carcinomas probably have a different pathogenesis from nasopharyngeal lymphoepithelioma. PMID- 2546459 TI - Lymphoepithelioma-like carcinoma of the lung. AB - Originally described in the nasopharynx, lymphoepitheliomas are undifferentiated carcinomas with prominent lymphoid infiltration. Recently this tumor has been described in a number of other sites. An association with prior infection by the Epstein-Barr virus has been recognized in cases from the nasopharynx. We report four cases of lymphoepithelioma-like carcinoma arising in the lung. Epstein-Barr virus serology indicated prior infection in two cases, one of which was also found by in situ hybridization to have the virus genome. Lymphoepithelioma-like carcinoma must be considered in the differential diagnosis of primary lung tumors, particularly when an extensive infiltrate by lymphoid tissue is present and may even suggest lymphoma. PMID- 2546460 TI - Correlation of histology and human papillomavirus DNA detection in condyloma acuminatum and condyloma-like vulvar lesions. AB - The diagnosis of a vulvar condyloma is made when perinuclear halos are seen with nuclear atypia and binucleate forms (koilocytotic atypia). These changes are most prominent in the granular layer and are associated with the presence of human papillomavirus (HPV). However, these changes may be absent or minimal in patients with papillary vulvar lesions; this situation can thus present diagnostic difficulties. We analyzed the histologic features of 53 biopsies from 48 patients who had vulvar lesions suggestive of condylomata. Of the 26 biopsy specimens with koilocytotic atypia, 20 (77%) had sequences homologous to HPV DNA as detected by Southern blot hybridization analysis using a probe of HPV s 6/11, 16, 18, 31, 35, and 51. In cases where the histologic features were suggestive but not diagnostic of condylomata, because unequivocal koilocytotic atypia was not noted, five of 27 (19%) had detectable HPV DNA. In this latter group, we found no histologic feature to distinguish the cases that had detectable HPV DNA from those that did not. Analysis for HPV DNA by in situ hybridization in the cases that were histologically equivocal for condyloma was uniformly negative. We conclude that there is a marked decrease in the detection rate of the HPV types associated with genital tract neoplasms in vulvar lesions that lack koilocytotic atypia. Southern blot hybridization analysis was the only reliable way to distinguish the "equivocal for condyloma" cases that had HPV from those where HPV DNA was not detected. PMID- 2546461 TI - Chronic ACTH treatment increases striatal dopamine D-2 receptor binding in developing rat brain. AB - ACTH has been reported to decrease elevated levels of dopamine metabolites in the CSF of patients with infantile spasms who respond clinically to ACTH therapy. To study the possible role of dopamine receptors in the effect of ACTH, we treated rat pups for thirty days with 40 IU/kg subcutaneously of porcine ACTH or with normal saline. Using 10 nM 3H-spiperone and sulpiride to determine nonspecific binding, specific binding of D-2 receptors increased significantly (46%) in the striata of ACTH-treated rats when compared to controls. No significant difference in specific binding was found in the nucleus accumbens. Protein concentration was significantly decreased by ACTH treatment. Saturation studies will be necessary to determine if the increase in dopamine receptor binding induced by a high dose of ACTH represents a change in receptor density or affinity. The effect of lower clinical doses of ACTH on dopamine receptors warrants study. PMID- 2546462 TI - Acquired immune deficiency, myelodysplasia, and acute nonlymphocytic leukemia associated with monosomy 7 and t(3;3) (q21;q26) in a child with Langerhans cell histiocytosis. AB - A case of therapy-related myelodysplasia followed by acute nonlymphocytic leukemia in a 5-year-old child successfully treated for diffuse Langerhans cell histiocytosis is described. A syndrome of severe cell-mediated immune deficiency and persistent Epstein-Barr virus (EBV) infection coincided with the evolution of myelodysplasia. Specific abnormalities of chromosomes number 7 and 3 were associated with the onset of myelodysplasia and acute nonlymphocytic leukemia and are believed to be linked to the patient's immune dysfunction and compromised ability to contain viral infection. PMID- 2546463 TI - Secondary leukemia following successful treatment of Wilms' tumor. AB - Leukemia accounts for 15-20% of the secondary malignancies among survivors of Wilms' tumor. We report three patients who developed leukemia after the successful treatment of Wilms' tumor, each of whom demonstrates the importance of close long-term medical surveillance. The first patient developed Philadelphia chromosome positive chronic myelogenous leukemia (CML) 6 years after the diagnosis of Wilms' tumor. This is the second report of CML occurring after Wilms' tumor. The other two patients developed acute nonlymphocytic leukemia (ANLL) 3 and 18 years after successful treatment of Wilms' tumor. In one patient, the clinical manifestations were subtle, and in the other the latency period was the longest reported for secondary leukemia following Wilms' tumor. We conclude that survivors of childhood cancer require frequent medical surveillance even in their adult years. PMID- 2546464 TI - Parvovirus B19 infection associated with reticulocytopenia and chronic autoimmune hemolytic anemia. AB - A previously healthy 11-year-old white female presented with a Coomb's positive hemolytic anemia and reticulocytopenia. The patient was treated with prednisone (6.4 mg/kg/day) and had a stable hemoglobin with subsequent recovery of reticulocytes by 7 days. Bone marrow aspiration showed hypercellularity with arrest of red cell maturation. The patient's serum contained specific IgM and IgG antibodies to parvovirus B19. Our report confirms parvovirus B19 as a cause of reticulocytopenia at presentation in autoimmune hemolytic anemia. Immunosuppressive therapy with prednisone did not appear to significantly prolong the course of the B19 infection. PMID- 2546465 TI - Effect of acute ethanol on uptake of [3H]adenosine by rat cerebellar synaptosomes. AB - Many classes of CNS-acting drugs have been suggested to act at least partially via inhibition of adenosine uptake. Synaptosomal uptake of [3H]adenosine and the effect of acute ethanol on it were studied in a rat brain area known to be involved in the coordination and modulation of normal motor activity, the cerebellum. Uptake of [3H]adenosine was found to be linear with time (about 40 sec) and increasing concentrations (up to 1.5 microM) of adenosine. The uptake of [3H]adenosine was inhibited by dilazep (IC50 = 2.5 x 10(-7) M) in a dose dependent manner. Pharmacologically and/or toxicologically relevant concentrations of ethanol (2.5 to 100 mM) significantly inhibited the uptake of [3H]adenosine between 12 and 15%. Lineweaver-Burk plots indicated that both in vitro (25 mM) and in vivo (1.5 g/kg i.p.; 30 mM blood level) ethanol lowered Km as well as Vmax values for adenosine uptake to nearly the same extent. In the case of in vivo ethanol, no ethanol was present during the assay since synaptosome preparation would wash out residual ethanol. The results of the present study indicate possible membranal alterations by in vivo ethanol. It is concluded that the uptake of [3H]adenosine is inhibited by intoxicating concentrations of ethanol in vitro and by acute ethanol (1.5 g/kg) in vivo. This may partially explain the modulatory role of endogenous adenosine in ethanol induced motor disturbances. PMID- 2546466 TI - Alcohol exposure in utero results in diminished T-cell function and alterations in brain corticotropin-releasing factor and ACTH content. AB - The long-term teratogenic effects of prenatal ethanol exposure during the last week of gestation on immune responsiveness and levels of pituitary ACTH and hypothalamic corticotropin-releasing factor (CRF) were examined in Sprague-Dawley rats. Immune responsiveness was measured by T-lymphocyte proliferation in response to mitogenic stimulation with Con A (3 micrograms/ml) in spleen and thymus cells of 21-old-day male rats who were exposed to alcohol in utero. The proliferative response was 8-fold lower in spleen and twofold lower in thymus cells from alcohol-exposed animals compared to responses measured in control rats. Thymus weight was significantly smaller at birth in alcohol exposed males, but significantly larger at 21 days of age compared to controls. Alterations in the content of ACTH and CRF, hormones, known to be direct or indirect modulators of immune responsiveness, were also observed in alcohol exposed males. Hypothalamic content of CRF and pituitary content of ACTH were significantly lower in alcohol exposed males on postnatal Day 1, but hypothalamic ACTH content was significantly higher compared to controls. These results indicate that alcohol exposure during the last week of gestation can produce alterations of the hypothalamic-pituitary-adrenal function in addition to teratogenic effects on the immune system which have been previously observed only with a much longer alcohol exposure regimen. PMID- 2546467 TI - Use of thiol-terminal silanes and heterobifunctional crosslinkers for immobilization of antibodies on silica surfaces. AB - A procedure for covalent immobilization of functional proteins on silica substrates was developed using thiol-terminal silanes and heterobifunctional cross-linkers. Using this procedure, a high density of functional antibodies was bound to glass cover slips and silica fibers. The amount of anti-IgG antibody immobilized was determined to be in the range of 0.66 to 0.96 ng/mm2 using radiolabeled antibody. The relative amount of IgG antigen bound by the immobilized antibody (0.37 to 0.55 mol antigen/mol antibody) was three to five times greater than other investigators have reported. In addition, the amount of protein nonspecifically adsorbed to the antibody-coated surface was further reduced by the addition of blocking agents so that nonspecific adsorption of protein antigens represented only 2-6% of the total antigen binding. With this low background, IgG antigen binding could be measured at levels as low as 150 fmol when an antigen concentration of 3 pmol/ml was applied. The process for antibody immobilization is straightforward, easy to perform, and adaptable for modifying mass quantities of biosensor components. PMID- 2546468 TI - Evidence of PAF-acether metabolic pathway activation in antigen challenge of upper respiratory airways. AB - Lyso-PAF-acether and PAF-acether (formerly platelet-activating factor) were detected in nasal secretions from patients with hay fever who underwent local antigen challenge. Lyso-PAF release was observed in 12 of 13 patients, with a maximum (p less than 0.001) 5 min after stimulation and a progressive decrease during the first hour. PAF was detected in the 5-min postchallenge nasal washings from two of 13 subjects. After HPLC, this mediator was found in four of seven postchallenge nasal washings submitted to this procedure, with a peak 5 min and 10 min after provocation. Histamine analysis revealed a significant (p less than 0.001) but time-limited (5 min) release in nasal secretion. The pattern of immunoreactive leukotriene C4 showed a maximal peak (p less than 0.01) 5 min after allergen provocation, with raised levels for 20 min. Nasal stimulation with nebulized saline solution or grass pollens in healthy subjects and in patients suffering from allergic rhinitis caused by Dermatophagoides pteronyssinus was followed by no local mediator release. These data indicate that, in addition to histamine and peptide-leukotrienes, lyso-PAF and PAF are released in nasal secretions after local antigen stimulation in patients with hay fever, with a preponderance of lyso-PAF response. On the basis of these results, it is conceivable that these ether-phospholipids may be involved in allergic inflammation of human nasal airways. PMID- 2546469 TI - Airway responsiveness to histamine and leukotriene E4 in subjects with aspirin induced asthma. AB - Airway responsiveness to histamine and leukotriene E4 (LTE4) has been compared between five subjects with aspirin-induced asthma (AIA) and 15 asthmatic subjects without aspirin sensitivity (non-AIA). In the AIA group, the geometric mean doses of histamine and LTE4 causing a 35% fall in specific airway conductance (PD35) were 0.31 mumol and 0.17 nmol, respectively, and LTE4 was 1,870 times more potent than histamine. In the non-AIA group, the histamine and LTE4 PD35 doses were 0.40 mumol (non-AIA versus AIA, NS) and 2.8 nmol (non-AIA versus AIA, p = 0.002), respectively, and LTE4 was 145 times more potent than histamine in eliciting bronchoconstriction (non-AIA versus AIA, p = 0.001). After desensitization to aspirin the geometric mean histamine and LTE4 PD 35 in the AIA group changed to 0.19 mumol (NS) and 3.3 nmol (p = 0.007), respectively, and there was an average 33-fold reduction in the responsiveness of the airways to LTE4 relative to histamine (p less than 0.001). In five non-AIA subjects. Ingestion of 600 mg of aspirin daily did not lead to any significant change in airway responsiveness to histamine or to LTE4. These results demonstrate a selective and marked increase in airway responsiveness to LTE4 in subjects with AIA. The efficacy of desensitization may relate in part to a selective down-regulation of LTE4 receptors within the airways. PMID- 2546470 TI - Effect of endothelial injury on the responses of isolated guinea pig pulmonary venules to reduced oxygen tension. AB - The influence of the endothelium on pulmonary venular responses to reduced oxygen tension has not been defined. To examine this question, endothelial injury was induced in small guinea pig pulmonary artery and venule segments (effective lumen radius, 174 +/- 5 and 122 +/- 2 microns, respectively) by perfusion with either a mixture of hypoxanthine (5 mM) and xanthine oxidase (0.05 U/ml) (HX/XO) or collagenase (2 mg/ml). HX/XO significantly (p less than 0.05) reduced the relaxation of precontracted pulmonary arteries by acetylcholine (ACH), bradykinin (BK), and A-23187, and the relaxations were restored by including superoxide dismutase (40 micrograms/ml) in the HX/XO solution. However, neither HX/XO nor collagenase affected vasodilation induced by ACH, BK, and A-23187 in precontracted pulmonary venules. In contrast, HX/XO significantly (p less than 0.05) augmented the sustained contraction of pulmonary venules to hypoxia (HX/XO, 3.2 +/- 1.0 mg/mm; control, 1.0 +/- 0.5 mg/mm) and anoxia (HX/XO, 35.1 +/- 6.6 mg/mm; control, 20.3 +/- 4.0 mg/mm). Collagenase also significantly (p less than 0.05) enhanced the anoxic contractions (collagenase, 36.0 +/- 3.7 mg/mm; control, 20.9 +/- 6.8 mg/mm). Superoxide dismutase (40 micrograms/ml) and catalase (323 micrograms/ml) abolished HX-XO-induced augmentation of the hypoxic and anoxic contractions of pulmonary venules. Collagenase removed 54 +/- 8% of the venular endothelium (control, 5 +/- 1%), whereas HX/XO-exposed endothelial cells contained numerous craters. Neither gossypol (5 microM) nor methylene blue (10 microM) affected pulmonary venular contractions to reduced PO2. Endothelial damage augments the PO2-dependent contractions of the pulmonary venule, and this augmentation does not appear to be due to decreased release of endothelium derived relaxing factor. PMID- 2546471 TI - The National Tuberculosis Training Initiative. PMID- 2546472 TI - CD4 counts as predictors of opportunistic pneumonias in human immunodeficiency virus (HIV) infection. AB - STUDY OBJECTIVE: To determine if circulating CD4+ lymphocyte counts are predictive of specific infectious or neoplastic processes causing pulmonary dysfunction. DESIGN: Retrospective, consecutive sample study. SETTING: Referral based clinic and wards. PATIENTS: We studied 100 patients infected with human immunodeficiency virus (HIV) who had had 119 episodes of pulmonary dysfunction within 60 days after CD4 lymphocyte determinations. MEASUREMENTS AND MAIN RESULTS: Circulating CD4 counts were less than 0.200 X 10(9) cells/L (200 cells/mm3) before 46 of 49 episodes of pneumocystis pneumonia, 8 of 8 episodes of cytomegalovirus pneumonia, and 7 of 7 episodes and 19 of 21 episodes of infection with Cryptococcus neoformans and Mycobacterium avium-intracellulare, respectively. In contrast, circulating CD4 counts before episodes of nonspecific interstitial pneumonia were quite variable: Of 41 episodes, 11 occurred when CD4 counts were greater than 0.200 X 10(9) cells/L. The percent of circulating lymphocytes that were CD4+ had a predictive value equal to that of CD4 counts. Serum p24 antigen levels had no predictive value. CONCLUSIONS: Pneumocystis pneumonia, cytomegalovirus pneumonia, and pulmonary infection caused by C. neoformans or M. avium-intracellulare are unlikely to occur in HIV-infected patients who have had a CD4 count above 0.200 to 0.250 X 10(9) cells/L (200 to 250 cells/mm3) or a CD4 percent above 20% to 25% in the 60 days before pulmonary evaluation. Patients infected with HIV who have a CD4 count below 0.200 X 10(9) cells/L (or less than 20% CD4 cells) are especially likely to benefit from antipneumocystis prophylaxis. PMID- 2546473 TI - Carcinogen-induced mutagenesis in the simian virus 40 genome. AB - Here are reviewed the most interesting results which have been obtained with a mutational assay based on the use of Simian Virus 40 (SV40) as a biological probe. This mutational assay allowed us first to study the mutation potency of some chemical and physical DNA damaging agents such as acetoxy acetylaminofluorene and UV-light and of apurinic sites created by heat treatment under acidic conditions, and second to study at the molecular level the modifications induced by these treatments. A correlation between the location of the DNA adducts and the location of the hot spots of mutagenesis has tentatively been researched. No direct link has been found. Our results suggest that mutation hot spots are correlated with local DNA conformations which could be modified by the DNA damaging agents. PMID- 2546474 TI - Biochemical and enzymatic aspects of caffeine and caffeine derivatives induced DNA strand breaks. AB - The results presented here point to the difficulties that exist in connection with the identification of the molecular target of caffeine. Our data support the evidence that caffeine and caffeine derivatives cause DNA-protein cross-links (DPC) in whole mammalian cells or in isolated nuclei. These DPC have the same properties (saturability, reversibility and temperature-dependence) as those produced by an enzymatic inhibition. The experiments performed in reconstituted systems, in the presence of purified DNA topoisomerase II, do not support the original hypothesis that this enzyme might be a possible target for this class of drugs. We suggest the possibility that other DNA metabolism enzymes are involved in the biological effects of caffeine and caffeine derivatives. Further biochemical and molecular data are necessary to identify which of these enzymes is in fact affected. PMID- 2546475 TI - Molecular mechanisms of hydrogen peroxide cytotoxicity. AB - The molecular mechanisms of H2O2 toxicity have been investigated in both mammalian or bacterial cells. DNA breakage mediates cytotoxicity by low concentrations of H2O2 in mammalian cells, but DNA lesions do not appear as a direct consequence of the action of the hydroxyl radical; rather, these radicals may disturb intracellular Ca2+ homeostasis, which results in secondary reactions ultimately leading to DNA strand breakage and cytotoxicity. Studies that have used Escherichia coli (E. coli) as a cellular system have indicated that the two modes of killing detectable in cells exposed to increasing concentrations of H2O2 are mediated by different radical species. Mode-one killing seems to be produced by the superoxide anion whereas mode-two killing seems to be a consequence of the hydroxyl radical attack. PMID- 2546476 TI - [Cancer of the maxillary sinus. A retrospective study of 29 cases]. AB - Representing 2% of malignant tumors and 6% of head and neck cancer, almost always diagnosed at a late stage, maxillary sinus cancer still remains a rather rare pathology but fearsome prognosis. From the 29 cases, over 10 years of follow up at the clinic and Paul Strauss cancer treatment center, this study focuses mainly on two criteria: diagnosis, in the study of the presenting symptoms in order to treat this pathology as soon as possible; therapeutic, by analysing our results, and by focusing on the current treatment. PMID- 2546477 TI - Preoperative and postoperative considerations in elective breast operations. AB - The number of young women undergoing elective breast operations is increasing rapidly. The lifetime breast cancer risk in the female population is 1 in 10. As this group ages, it is obvious that surgeons will encounter carcinoma of the breast in these patients more frequently. The surgical implications of breast cancer in these patients are discussed, and guidelines for preoperative and postoperative management in patients being considered for augmentation mammaplasty, mastopexy, and reduction mammaplasty are suggested. PMID- 2546478 TI - [Syncytia formation in cultures and analysis of the protein composition of various strains of caprine arthritis encephalitis virus (CAEV)]. AB - The syncytia forming activity of 9 caprine arthritis encephalitis virus (CAEV) strains isolated from lesions or normal goat tissue was evaluated in vitro. No discrepancies were seen between the structural proteins of the 9 strains by gel electrophoresis. Goat synovial membrane cells were the most susceptible to the 9 CAEV strains. Goat monocytes in culture were susceptible to all the strains tested. At 39 degrees C, all the strains produced more syncitial lesions. In nasal turbinate cells, 2 strains did not form syncytia; one strain induced lysis of all the goat cells used. PMID- 2546479 TI - [Detection of visna-maedi virus antibodies and antigens by immunoblotting]. AB - The visna-maedi virus was immunologically diagnosed using an immunoblotting technique from an antigenic preparation of the visna-maedi virus K796 purified by sucrose density gradient centrifugation. After SDS-PAGE electrophoresis and transfer onto a nitrocellulose sheet, the immunoblotting procedure was adapted to the search for specific antibodies in ovine serum samples. The results obtained showed that, in the natural visna-maedi disease, antibodies are not systematically detectable against every viral protein of the virus core. We have demonstrated the existence of antibodies directed against the proteins coded by the gag and the pol genes. PMID- 2546480 TI - Comparison of fluorescence polarization immunoassay, enzyme immunoassay, and thin layer chromatography for urine cannabinoid screening. Effects of analyte adsorption and vigorous mixing of specimen on detectability. AB - Four commercial assays for the screening of cannabinoids in urine were compared. Urine specimens from 93 selected subjects were run by fluorescence polarization immunoassay on the Abbott TDx; by enzyme multiplied immunoassay with two Syva EMIT assays; and by thin-layer chromatography with the TOXI-LAB system (Marion Laboratories). The TDx cannabinoid threshold can be set anywhere from 25 to 150 micrograms per L. Twenty-five micrograms per L was chosen for this study. The thresholds for EMIT are fixed at 20 micrograms per L for one assay and 100 micrograms per L for the other. The detection limit for TOXI-LAB, according to the manufacturer, can be anywhere from 5 to 50 micrograms/L, depending on the specimen. Urines, positive by at least one method, were further analyzed by gas chromatography with mass spectrometry (GC/MS), The detection limit for the GC/MS method was 10 micrograms per L. The results showed a few false negatives and unconfirmable positives; in general, correlation was considered acceptable. Dose response curves comparing TDx and EMIT gave paralell results, with comparable cross-reactivity for the major metabolite, 11-nor-delta-9-tetrahydrocannabinol-9 carboxylic acid (delta-9-THC-COOH). A dose-response study of TOXI-LAB using delta 9-THC-COOH also gave acceptable results. Adsorption to glass was investigated using spiked urine; a 27 percent reduction in concentration was caused by this phenomenon. Foaming of spiked urine caused by vigorous mixing resulted in a reversible 89 percent apparent reduction in concentration. PMID- 2546482 TI - Clinical pharmacology of gallium chloride after oral administration in lung cancer patients. AB - Pharmacokinetic parameters were determined in 18 lung cancer patients after a single administration of 800 mg/24 h of GaCl3: Cmax = 123 +/- 61 mu/l; Tmax = 5.2 +/- 5.5 h; AUCO-96h = 4690 +/- 3358 micrograms.l-1.h; AUCO - infinity = 6394 +/- 5352 micrograms.l-1.h; T 1/2 beta = 43 +/- 19 h. Serum Ga concentrations at the steady-state (Css) were then determined in these patients after a daily oral administration of 800 mg/24 h of GaCl3 for 15 days: Css = 274 +/- 167 micrograms/l. No correlation was found between Css and the previous pharmacokinetic parameters in each patient. Various doses of GaCl3 were administered daily to 45 patients to correlate Css and dosage. Serum Ga concentrations increased with dosage from 100 to 400 mg/24 h (p less than 0.05), but not with further dosages up to 1400 mg/24 h. The optimal daily dose of GaCl3 in lung cancer patients seems to be 400 mg/24 h. In 2 patients, Ga was assayed after death in tissues. Ga concentrations were more than 10 micrograms/g in metastases, 3.6 +/- 2.9 micrograms/g in the primary tumor and 2.3 +/- 0.9 micrograms/g in the kidney. Due to the lack of renal and hematological toxicities and the significant uptake of Ga by the tumor, GaCl3 can be used orally in conjunction with other cytotoxic agents. We intend to evaluate its efficacy according to a randomized study comparing chemotherapy versus chemotherapy plus 400 mg/24 h of GaCl3. PMID- 2546481 TI - Inhibition of herpes simplex virus infection by pine cone antitumor substances. AB - Several antitumor substances extracted from cones of various pine trees inhibited the plaque formation of Herpes simplex virus types 1 and 2 (HSV-1, HSV-2) strains in African green monkey kidney cells and human adenocarcinoma cells. The 50% effective dose of the most active fraction, Fr. VI (0.3 micrograms/ml) was 0.001 times its 50% cytotoxic dose (greater than 300 micrograms/ml). The anti-HSV activity of various pine cone extracts increased with their acidity. Identification of the polyphenol groups as donors of the acidity was further supported by the observation that the anti-HSV activity of the natural polyphenolic products, such as tannin and lignin, significantly exceeded that of other natural or chemically modified antitumor polysaccharides. An experiment using radiolabeled virus particles indicated that the anti-HSV effect of both Fr. VI and lignin was attributable to interference with virus adsorption to these cells rather than to inhibition of virus penetration into the cells. PMID- 2546484 TI - Intrapleural BCG in postsurgical stage I non-small cell lung cancer. AB - In a randomized trial comparing intrapleural bacillus Calmette-Guerin (BCG) with placebo in postsurgical stage I non-small cell lung cancer, BCG-treated patients (n = 17) showed a non-significant overall improvement of 5-year and median survival and disease-free interval, but an increased rate of recurrence of cancer compared with control patients (n = 17). PMID- 2546483 TI - Malignant transformation of host cells by a human small cell lung cancer xenografted into nude mice. AB - Malignant transformation of mouse host cells by a human small cell lung cancer (SCLC) was demonstrated by short-term in vitro cultivation of the tumor cells from a xenograft at two different transplant generations. Isoenzyme (LDH) and chromosome analysis showed that out of the 3 cell lines established from this tumor, 1 retained a human karyotype similar to that of the xenograft and 2 were murine transformed cell lines. These murine cell lines produced fibro-sarcoma like tumors when injected into nude mice. Because of the early in vitro emergence of murine transformed cell populations, it is likely that the transformation process had occurred in vivo. Since in our experience the induction of transformation of host murine cells, also observed directly in vivo, is more frequent with SCLC than other histotypes (lung and colorectal adenocarcinoma), it is suggested that the known production of growth factor by these tumors may contribute to this transformation. PMID- 2546485 TI - Multicellular aggregates from human tumor cell lines for radiation studies. AB - Aggregates of human tumor cells are widely used in experimental studies on tumor responses to treatment. Only a limited number of human tumor cell lines are capable of forming spheroids. In this study cellular characteristics of 7 lung cancer and 4 bladder cancer cell lines are described with respect to their spheroid forming capacity. Comparisons were made with four reference lines known for their propensity to form growing aggregates. In the absence of vimentin expression no spherical aggregates were formed. Spherical aggregates were formed by one bladder and one lung cancer cell line, of which only the latter exhibited growth. Cellular factors influencing the ability of spheroids to increase in volume after spherical aggregation are not yet defined. Viability and clonogenicity of cells in aggregates are not the determinant of growth capacity. The growth rate of cell lines that exhibited growth is determined by tissue culture conditions and additives. Type of medium, percentage of foetal bovine serum and glucose concentration influenced the growth rate of spheroids. Since the response to radiation may be influenced by the growth rate of the tumors, manipulation of tissue culture medium composition offers the possibility of testing the influence of growth rate on the radiation response of one type of spheroids. PMID- 2546486 TI - Biological and biochemical characterization of clinical isolates of herpes simplex virus type 2 resistant to acyclovir. AB - A series of clinical isolates of herpes simplex virus type 2 were taken from a patient with chronic lymphocytic leukemia. Acyclovir (ACV) susceptibility assays revealed that some isolates were resistant to ACV and cross-resistant to ganciclovir but not to phosphonoacetic acid. The nature of the resistance was examined further. A number of cloned variants were generated, and thymidine kinase and DNA polymerase assays were carried out. Variants that were resistant to ACV were found to be thymidine kinase deficient. Evidence for alteration in the DNA polymerase was not found when ACV triphosphate or phosphonoacetic acid was used as the inhibitor. In vivo studies with the plaque-purified viruses showed that ACV resistance was associated with a reduced neurovirulence. In a zosteriform model, virus resistant to ACV was unable to induce secondary spread in the same dermatome, to invade the peripheral nervous system or the central nervous system, or to establish latent infections. PMID- 2546487 TI - Combinations of 3'-azido-3'-deoxythymidine (zidovudine) and phosphonoformate (foscarnet) against human immunodeficiency virus type 1 and cytomegalovirus replication in vitro. AB - Combinations of 3'-azido-3'-deoxythymidine and phosphonoformate produced a moderate synergistic inhibitory effect against human immunodeficiency virus type 1 in vitro at concentrations that are easily achieved in humans. The synergistic effect was more pronounced with increasing concentrations and was not secondary to toxic effects of the drugs. 3'-Azido-3'-deoxythymidine neither inhibited the replication of human cytomegalovirus in human embryonic lung fibroblasts nor interfered with the anticytomegalovirus effect of phosphonoformate. By using partially purified reverse transcriptase of human immunodeficiency virus type 1 and human cytomegalovirus DNA polymerase, various combinations of 3'-azido-3' deoxythymidine-5'-triphosphate and phosphonoformate produced strong indications of additive interactions. The synergistic interactions in infected cells and the additive effects observed at the reverse transcriptase level indicate that mechanisms other than the reverse transcriptase may be of importance for the inhibition of human immunodeficiency virus replication by these two compounds. A concomitant treatment of cytomegalovirus infections, such as cytomegalovirus retinitis, with phosphonoformate in patients with acquired immunodeficiency syndrome receiving 3'-azido-3'-deoxythymidine may be appropriate, and this combination may also be useful in controlling human immunodeficiency virus infection. PMID- 2546488 TI - Comparative evaluation of daptomycin (LY146032) and vancomycin in the treatment of experimental methicillin-resistant Staphylococcus aureus osteomyelitis in rabbits. AB - A rabbit model for methicillin-resistant Staphylococcus aureus (MRSA) osteomyelitis was used to compare treatment with daptomycin, a new peptolide, and vancomycin. Daptomycin (4 mg/kg) and vancomycin (40 mg/kg) were injected subcutaneously every 12 and 6 h, respectively. After treatment, MRSA was found in bone cultures from 18 of 18 control rabbits, 10 of 17 animals treated with daptomycin, and 11 of 18 animals treated with vancomycin. Drug concentrations were measured in serum, uninfected bone, and infected bone 1 h after daptomycin or vancomycin was injected in a group of rabbits that had been infected for 3 to 4 weeks. Vancomycin was present at the highest concentrations in infected and uninfected bone. The results of this study suggest that daptomycin was similar to vancomycin in the eradication of MRSA from infected bone in an experimental model of osteomyelitis. PMID- 2546489 TI - Assay of fluconazole by megabore capillary gas-liquid chromatography with nitrogen-selective detection. AB - A megabore column gas-liquid chromatographic method which uses nitrogen phosphorus detection was developed for the analysis of fluconazole in plasma, serum, cerebrospinal fluid, or urine. The assay was linear from 0.2 to 200 micrograms/ml and had an average coefficient of variation of 7%. The suitability of the assay for pharmacokinetic studies was demonstrated. PMID- 2546490 TI - Foscarnet treatment of cytomegalovirus retinitis in patients with the acquired immunodeficiency syndrome. AB - Ten patients with acquired immunodeficiency syndrome with newly diagnosed cytomegalovirus (CMV) retinitis were treated with an induction regimen of intravenous foscarnet, 60 mg/kg of body weight, administered as a 2-h infusion and repeated every 8 h for 14 days. At the end of induction, 9 of 10 patients had stabilized (no new retinal lesions and stable old lesions [7 patients]) or improved (decreased retinal opacification [2 patients]). All eight patients with CMV in urine or blood upon entry into the study had negative urine and blood cultures at the end of induction. After induction therapy, seven patients continued maintenance foscarnet therapy, 60 mg/kg as a single daily infusion, 5 days/week. In six patients, retinal lesions increased in size after 2 to 32 weeks of maintenance therapy. One was invaluable because a retinal detachment developed. Only 9 of 42 blood and urine cultures obtained during maintenance foscarnet therapy yielded CMV, compared with 7 of 14 obtained prior to the initiation of foscarnet induction therapy (P = 0.04). Foscarnet toxicity was mild and infrequent: elevation in serum creatinine by 0.5 to 1.3 mg/dl over the base line (two patients), muscle twitching (three patients), hemoglobin decrease by 1 mg/dl (two patients), nausea (two patients), absolute neutrophil count decrease by 50% (one patient), rise in serum phosphorus to greater than 5.5 mg/dl (four patients), and proteinuria (two patients). Intermittently administered intravenous foscarnet appears to be an effective, relatively nontoxic therapy for CMV retinitis. Additional studies to determine the optimal dosage for maintenance therapy are needed, as are comparative trials with ganciclovir. PMID- 2546491 TI - Pharmacokinetics of intermittently administered intravenous foscarnet in the treatment of acquired immunodeficiency syndrome patients with serious cytomegalovirus retinitis. AB - Foscarnet has been shown to be active in vitro against the human immunodeficiency virus and all human herpesviruses including cytomegalovirus (CMV). A pharmacokinetic study was carried out as part of a clinical trial designed to evaluate the safety and efficacy of intermittently administered intravenous foscarnet for the treatment of CMV retinitis. Eight patients with acquired immunodeficiency syndrome and serious CMV retinitis received 2-h intravenous infusions of foscarnet at a dosage of 60 mg/kg of body weight every 8 h for 14 days. Serial plasma samples were collected on days 3 and 14 of therapy, and foscarnet concentrations were determined by high-pressure liquid chromatography. On day 3 of therapy, the mean (+/- standard deviation) peak and trough levels in plasma were 509 (200) and 98 (29) microM, respectively, while on day 14 levels were 495 (149) and 126 (59) microM. The mean clearance in plasma on days 3 and 14 were 1.9 (0.6) and 1.7 (0.9) ml/min per kg, respectively. On day 14, the mean half-life was 4.5 (1.2) h and the volume of distribution was 0.74 (0.60) liter/kg. As the half-life and the clearance of foscarnet in plasma correlated with changes in renal function, dosage adjustments must be made for patients with decreased renal function. PMID- 2546493 TI - Quinolone resistance in clinical isolates of Serratia marcescens. AB - The uptakes of norfloxacin by quinolone-resistant and -susceptible strains of Serratia marcescens were almost the same and 50% inhibitory concentrations for DNA gyrase and the MICs of quinolones were correlated, suggesting that DNA gyrase alterations are the basis of quinolone resistance. PMID- 2546494 TI - Epidural portals. Long-term management of intractable pain. PMID- 2546492 TI - Organization of complex transposon Tn2610 carrying two copies of tnpA and tnpR. AB - Transposon Tn2610 has two elements of 3.5 kilobase pairs as inverted repeats, one set at each end. This unique terminal element contained the transposition genes tnpA and tnpR. Only the tnpA gene in the left element was functional for transposition, whereas both tnpR genes were active. Possible evolutionary relationships among class II transposable elements are proposed on the basis of the genetic and structural organization of Tn2610. PMID- 2546495 TI - Phosphorylation-dependent regulation of amidotransferase during the development of Blastocladiella emersonii. AB - The enzyme amidotransferase [2-amino-2-deoxy-D-glucose-6-phosphate ketol isomerase (amino-transferring); EC 2.6.1.16] catalyzes the first step in the hexosamine biosynthetic pathway. In Blastocladiella emersonii the sensitivity of the enzyme to the inhibitor uridine-5'-diphospho-N-acetylglucosamine (UDP-GlcNAc) is developmentally regulated. The inhibitable form of amidotransferase activity present in the zoospore is converted to a noninhibitable form during germination. The latter form is present throughout the growth phase and sensitivity to UDP GlcNAc gradually returns to the zoospore level during sporulation [C.P. Selitrennikoff, N.E. Dalley, and D.R. Sonneborn (1980) Proc. Natl. Acad. Sci. USA 77, 5998-6002]. The following evidence suggests that a phosphorylation/dephosphorylation mechanism underlies this interconversion: (i) Both the vegetative and zoospore enzymes have the same molecular weight of 140,000, but the vegetative enzyme elutes significantly earlier on a DEAE cellulose column than does the zoospore enzyme. (ii) The increased sensitivity to UDP-GlcNAc occurring in vivo and in vitro correlates with increased phosphorylation of a polypeptide of apparent Mr 76,000. This component copurifies with amidotransferase activity through ion-exchange chromatography and sucrose density gradient centrifugation. (iii) Desensitization and concurrent dephosphorylation of sensitive amidotransferase can be observed in vitro after treatment with a partially purified magnesium-dependent phosphoprotein phosphatase from zoospores. PMID- 2546496 TI - Catalytic activity of cytochrome oxidase and cytochrome c in apolar solvents containing phospholipids and low amounts of water. AB - Cytochrome c and cytochrome oxidase, in bovine heart submitochondrial particles and in their purified forms, were transferred to a ternary system that contained phospholipids (10 mg/ml toluene), the apolar solvent toluene, and water at concentrations of 13-15 microliters (high water) and 3 microliters (low water) per milliliter of toluene. When the enzymes were transferred back to an all water system, they exhibited full catalytic capacity. In the low water ternary system, cytochrome c could be reduced by ascorbate introduced via inverted micelles. Also in this system, cytochrome oxidase was reduced by ascorbate and cytochrome c but its oxidation was highly impaired. Data on the kinetics of reduction by ascorbate of cytochrome c and cytochrome oxidase under these conditions are presented. Cytochrome oxidase reduced in the organic solvent by ascorbate failed to form a complex with CO, but formed a complex with cyanide introduced via inverted micelles. The oxidized and the ascorbate-reduced cytochrome oxidase-cyanide complex exhibited a trough at 415 nm and a peak at 433 nm. The extent and rate of formation of the cyanide complex were higher with the reduced form of cytochrome oxidase. To achieve protein-protein interactions (cytochrome c-cytochrome oxidase) in the ternary system, it was necessary to extract the two proteins together. There was no functional interaction when they were extracted separately and mixed. In the high water ternary system reduced cytochrome oxidase was not detected, and it oxidized ascorbate at a higher rate than in the low water system; however, this rate was several orders of magnitude lower than in aqueous media. PMID- 2546497 TI - Induction of cytochrome P450-dependent monooxygenase activities in rat hepatoma H 4-IIE cells in culture by 2,3,7,8-tetrachlorodibenzo-p-dioxin and related compounds: mechanistic studies using radiolabeled congeners. AB - Treatment of rat hepatoma H-4-IIE cells in culture with 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD), 2,3,7,8-tetrachlorodibenzofuran (TCDF), 1,2,3,7,8-pentachlorodibenzo-p-dioxin (PeCDD), 1,2,3,7,8-pentachlorodibenzofuran (PeCDF), 1,2,7,8-TCDF, and 2,3,7-trichlorodibenzo-p-dioxin (TrCDD) resulted in the structure-dependent induction of aryl hydrocarbon hydroxylase and ethoxyresorufin O-deethylase activities. The induction potencies followed the order 2,3,7,8-TCDD greater than 2,3,7,8-TCDF greater than 1,2,3,7,8-PeCDD approximately 1,2,3,7,8-PeCDF greater than 1,2,7,8-TCDF greater than 2,3,7-TrCDD and were comparable to structure-toxicity relationships which have previously been reported. In contrast, many of the properties of these compounds were structure-independent. For example, using tritiated congeners of high specific activity (greater than 30 Ci/mmol) the sedimentation coefficients (S) for the nuclear and cytosolic aryl hydrocarbon (Ah) receptor complexes were 5-6 and 9-10 S, respectively, for all the radioligands. Moreover, examination of the processing of nuclear Ah receptor complexes for the radiolabeled congeners showed that after 6 h, the rates of nuclear processing were very low and varied between 0.006 and 0.0385 fmol degraded/mg protein/mg total DNA. These results were consistent with the reported stability and persistence of the nuclear Ah receptor complexes and in addition, there were no apparent structure-dependent differences in the processing rates. Inspection of the nuclear receptor levels and the corresponding induced enzyme activities for the congeners showed that there was a linear correlation between average nuclear receptor complex levels (18-42 h) and induced enzyme activities (32-42 h) for all six radioligands; these data indicated that the rates of cytochrome P450-dependent gene expression correlated with the levels of nuclear Ah receptor complex. In contrast, the accumulation of occupied nuclear receptor complexes in rat hepatoma H-4-IIE cells was structure dependent and appeared to be one of the factors which governed the observed structure-induction and the previously reported structure-toxicity relationships for 2,3,7,8-TCDD and related halogenated aryl hydrocarbons. PMID- 2546498 TI - Augmentation of inorganic pyrophosphate elaboration in cartilage by serum factors. AB - The disordered production of inorganic pyrophosphate (PPi) by articular cartilage is thought to have an important role in the pathogenesis of calcium pyrophosphate dihydrate deposition disease and perhaps osteoarthritis. We have previously shown that fetal calf serum added to the culture media of porcine articular cartilage explants increases the elaboration of PPi into the ambient media. We have examined this PPi stimulatory activity by studying the effects of adult human serum (HS), serum derived from adult human plasma (HP), and an acid-alcohol extract of human platelets (PE) on PPi production in cartilage organ culture. Ten percent HS produces a 1.4-fold increase in PPi production after 48 h of culture, while cartilage incubated in media containing 10% HP produces no more PPi than that incubated in media alone. PE stimulates a mean 2-fold increase in PPi production at 48 h in the presence of low concentrations of HP, and has no effect alone. It does not appear to up-regulate the activity of the ectoenzyme nucleoside triphosphate pyrophosphohydrolase (NTPPPH), nor does it promote the release of enzyme substrate into the extracellular space. Cartilage exposed to 0.5% HP and PE has 1.51 +/- 0.36 units of NTPPPH activity whereas cartilage exposed to 0.5% HP alone has 1.52 +/- 0.41 units of enzyme activity. PE does not increase the release of [14C]adenine-labeled compounds into the media. Approximately 13% of soluble 14C counts was found in the media of chondrocytes treated with PE while 18% of counts was released in the presence of HP alone. We have demonstrated a factor or factors present in FCS, HS, and an acid-ethanol extract of human platelets which represent(s) the first known physiologic modulators of PPi production in articular cartilage and may increase PPi production without affecting NTPPPH activity. PMID- 2546499 TI - Stimulation of inorganic pyrophosphate elaboration by cultured cartilage and chondrocytes. AB - Inorganic pyrophosphate elaboration by articular cartilage may favor calcium pyrophosphate dihydrate crystal deposition. Frequently crystal deposits form in persons affected with metabolic diseases. The cartilage organ culture system was used to model these metabolic conditions while measuring the influence on extracellular pyrophosphate elaboration. Alterations of ambient pH, thyroid stimulating hormone levels, and parathyroid hormone levels did not change pyrophosphate accumulation in the media. However, subphysiologic ambient calcium concentrations (25, 100, 500 microM) increased pyrophosphate accumulation about chondrocytes 3- to 10-fold. Low calcium also induced release of [14C]adenine labeled nucleotides from chondrocytes, potential substrates for generation of extracellular pyrophosphate by ectoenzymes. Exposing cartilage to 10% fetal bovine serum also enhanced by 50% the egress of inorganic pyrophosphate from the tissue. PMID- 2546500 TI - A model for hydropathy-based peptide interactions. AB - Two peptides are specified when the noncoding DNA strand is read in the 5' to 3', or the 3' to 5' direction, and both peptides form strong complexes with the natural peptide, as found by J. E. Blalock and K. L. Bost with ACTH [1986) Biochem. J. 234, 679-683). We report here that strong hydropathic complementarity (pairing of hydrophobic with hydrophilic residues), the assumed basis of these interactions, is obtained only if the peptide resulting from reading in the 3' to 5' direction is aligned parallel to the natural peptide, or if the peptide derived by opposite reading of the DNA is aligned antiparallel to it. Complementary is abolished in other alignments, including all staggered ones. In the appropriate alignments of the constructs the amino acid residues opposite one another are specified by a pair of complementary codons in the DNA; Blalock and Bost have indeed shown that complementary pairs of codons specify amino acids of opposite hydropathy. A model is proposed to explain how hydropathic complementarity can lead to interaction between peptides. We propose that in the interacting peptides hydrophilic residues of both chains are oriented toward the aqueous solvent, while the hydrophobic ones form the interphase between the two chains. Tight packing is made possible by the stipulation that whenever a hydrophilic residue turns toward the aqueous phase, a space is liberated which can accommodate a hydrophobic residue from the opposing chain. This entropy driven configuration can lead to strong interactions between portions of peptides consisting of hydropathically complementary residues. PMID- 2546501 TI - Ketoconazole inhibits self-induced metabolism of 1,25-dihydroxyvitamin D3 and amplifies 1,25-dihydroxyvitamin D3 receptor up-regulation in rat osteosarcoma cells. AB - Ketoconazole (an inhibitor of vitamin D-24 hydroxylase) was used to study the role of self-induced 1,25-dihydroxyvitamin D3 (1,25-D3) metabolism on cellular responsiveness to 1,25-D3. Eighteen hours of treatment with 1,25-dihydroxy-[26,27 methyl-3H]vitamin D3 (1,25-[3H]D3) increased total 1,25-D3 receptors (VDR) from 60 to 170 fmol mg/protein. In cells treated with both 1,25-[3H]D3 and ketoconazole, up-regulation of VDR was increased by 40% over that observed with cells receiving 1,25-[3H]D3 alone. Ketoconazole alone had no agonistic activity. Treatment of cells with 1 nM 1,25-[3H]D3 plus increasing doses of ketoconazole (0 30 microM) resulted in a dose-dependent increase in occupied VDR and total VDR. This up-regulation was associated with reduced 1,25-[3H]D3 catabolism. 1,25 [3H]D3-induced up-regulation of VDR typically peaked at 14 h and declined thereafter. Ketoconazole lengthened the time to reach peak VDR up-regulation to 20 h. The ability of ketoconazole to increase cell responsiveness (VDR up regulation) was the result of both increased and prolonged occupancy of VDR by 1,25-[3H]D3. The t1/2 of occupied VDR was 2 h in the absence of ketoconazole and greater than 7 h when ketoconazole was present. Collectively, these results suggested that self-induced catabolism of 1,25-D3 is an important regulator of VDR occupancy and therefore cellular responsiveness to hormone. These data also demonstrate the usefulness of ketoconazole as an inhibitor of vitamin D hydroxylases in intact cells. PMID- 2546502 TI - Analysis of the regulation of phosphatidylinositol-4,5-bisphosphate synthesis by arachidonic acid in exocrine pancreas. AB - In pancreatic acinar cells prelabeled with either 32Pi or myo-[3H]inositol, arachidonic acid (10-50 microM) rapidly decreased the steady-state levels of [32P]phosphatidylinositol 4',5'-bisphosphate [( 32P]PtdIns4,5P2) and inhibited carbachol-stimulated accumulation of [3H]inositol trisphosphate [( 3H]InsP3). Both actions of arachidonic acid were rapidly reversed by bovine serum albumin (BSA). Indomethacin and nordihydoguaiaretic acid failed to block the inhibitory effects of arachidonic acid on [32P]PtdIns4,5P2 levels. Arachidonic acid (10-50 microM) also caused a prompt depletion of cellular ATP which was rapidly reversed by BSA. The ATP-depleting action of arachidonate paralleled in terms of concentration dependence and time course its inhibitory effects on [32P]PtdIns4,5P2 and [3H]InsP3 levels. Exposure of acinar cells to 50 microM arachidonic acid produced an increase in oxygen consumption which exceeded that elicited by either carbachol or ionomycin. Arachidonic acid (10-50 microM) also caused a concentration-dependent rise in cytosolic Ca2+, which was partially obtunded by Ca2+ deprivation. A proposed mechanism involving arachidonic acid as a negative feedback regulator of polyphosphoinositide turnover in exocrine pancreas is discussed. PMID- 2546503 TI - [Phase I and pharmacokinetic study of KRN8602, a new morpholino anthracycline]. AB - Phase I clinical trial of a new semi-synthetic morpholino anthracycline derivative, KRN8602, was performed. Sixteen patients with advanced malignant neoplasms refractory to standard chemotherapies received 27 courses at doses ranging from 1.5 mg/m2/day to 18 mg/m2/day by bolus injection for three consecutive days. The dose limiting toxicity was leukopenia, and a maximally tolerated dose was 18 mg/m2/day (day 1-3). The recommended dose and schedule for a phase II study is determined to be 12 mg/m2/day for three consecutive days at 3 4 weeks intervals. Among non-hematologic toxicities, nausea and vomiting were severe, but stomatitis and alopecia were rarely observed. Clinical signs of cardiotoxicity were not seen. PMID- 2546504 TI - [Clinical study of combination chemotherapy with mitomycin C, vindesine and cisplatin (MVP therapy) in advanced non-small cell lung cancer]. AB - Thirty-seven patients with inoperable non-small cell lung cancer were treated with the combination chemotherapy (MVP therapy) with mitomycin C (8 mg/m2), vindesine (3 mg/m2 X 2) and cisplatin (60 mg/m2). The partial responders were 13 cases (35%), and the median survival time was 271 days. In this study the cisplatin dose was less than in any other report of "MVP" therapy. But both the response rate and the median survival time did not differ from those reported elsewhere. The side effects (bone marrow suppression, renal toxicity, etc.) were mild, and did not prevent the continuance of this therapy. Thus, we could repeat more than 6 courses of "MVP" therapy for 8 patients. Nowadays, it is difficult to obtain complete responders with any chemotherapy for inoperable non-small cell lung cancer. To prolong lives of patients and maintain good quality of life, we recommend chemotherapy with low toxicity in often-repeatable courses. PMID- 2546506 TI - [Pilot study of a continuous five-day intravenous infusion of cisplatin plus vindesine in advanced non-small cell lung cancer]. PMID- 2546505 TI - [Metastatic liver cancer from the stomach successfully treated by combined immunochemotherapy and transarterial embolization]. AB - A 53-year-old-man, who suffered from advanced gastric carcinoma with liver metastasis (P0H3S2N1; stage IV) underwent simple gastrectomy. After the operation, the patient was treated by chemotherapy (1/2 MFC, M: mitomycin C, F: FT-207, C: Cylocide) combined with immunotherapy (PSK, lentinan) and intraarterial injection (mitomycin C & Lipiodol). Liver metastases disappeared soon after the combined therapy, and these findings were confirmed by CT and US. Moreover, the serum level of CEA and CA 19-9 also decreased from 160 ng/ml and 51 U/ml to the normal level. The duration of the complete disappearance of the liver metastasis was not so long, but quality of life was well maintained for 2 and one half years. This case suggested that combined therapy may well be effective for advanced gastric cancer with liver metastases. PMID- 2546507 TI - [Effect of the pulmonary artery infusion chemotherapy of non-small cel lung cancer]. PMID- 2546508 TI - Does dexamethasone suppress the ACTH response in preterm babies? AB - Tests of adrenal stimulation with adrenocorticotrophic hormone (ACTH) were performed before and after commencing dexamethasone treatment in 12 infants. All except one of the tests performed showed the expected twofold rise in serum cortisol, suggesting that in this group of premature babies dexamethasone did not suppress the adrenal response. PMID- 2546510 TI - Ultraviolet irradiation for hepatic rickets. AB - An infant with chronic cytomegalovirus hepatitis and a child with atypical Alagille's syndrome had vitamin D deficiency rickets due to malabsorption. Both received ultraviolet irradiation. This treatment corrected biochemical abnormalities and healed the rickets. In the infant use of a sunlamp at home maintained normal 25 hydroxy-vitamin D for over a year. Our study shows that ultraviolet irradiation is an effective treatment of hepatobiliary rickets. PMID- 2546509 TI - Successful treatment after 'drowning' in sand. AB - A 30 month old boy who aspirated a large amount of dirty sand was successfully resuscitated and made a full recovery. PMID- 2546511 TI - Cocaine addiction: the hidden dimension. AB - There is growing awareness within the nursing profession that nurses need to expand their knowledge about addiction and develop expertise in providing care for substance abusing clients. This report presents a discussion about cocaine abuse that is focused on evolving knowledge about the physiology of addiction. Researchers have recently described cocaine-induced neurochemical changes in the brain that may form the underpinnings for the behavioral manifestations and symptomatology that have been associated with cocaine addiction. These neurochemical alterations are described at the cellular level, and treatment implications for nurses are presented. PMID- 2546512 TI - [Immunomodulating properties of interferon inducers]. AB - Data on the immunomodulating activity of interferon inductors are presented. It was revealed that the inductors increased the animal vaccinal response. Schemes for combined use of the interferon inductors and immunomodulators were developed. The immunomodulators were shown to increase the host interferon response evident from synergistic increasing of the interferon titers or prolongation of interferon circulation in blood of the animals. The efficiency of the schemes for combined use of the interferon inductors and immunomodulators was obvious from stimulation of the antibody production. As a result the time of the antibody circulation in blood increased. The effect of the combined use of the immunomodulators and interferon inductors was studied. The combined use of the preparations significantly increased the average life-span of the animals and the rate of their survival. PMID- 2546513 TI - [Neutralizing activity against herpes simplex in maternal milk]. AB - Human milk could play a role in the protection of the newborn from Herpes simplex virus 2 contamination. Neutralizing activity against Herpes simplex virus 2 was measured in 94 human milk samples and in 31 sera from healthy women. A weak neutralizing activity was detected in 48 milk samples. They were neither correlated to anti-Herpes simplex virus antibodies found in milk, nor to the neutralizing activity in sera. The origin and non immunologic nature of this activity are discussed. PMID- 2546514 TI - [The energy preservation requirement of growing pigs of different sexes fed normal and high protein diets. 4. Determination of parameters in the blood for estimating metabolic status]. AB - In connection with experiments for the ascertainment of energy and nutrient metabolization in castrated pigs, sows and boars over the complete fattening period and with alternating maintenance and growth feeding as well as the inclusion of an extreme protein supply level, blood parameters were studied of 3 animals per variant in order to recognize possible deviations in the metabolism and to be able to better assess the results of the measurements of energy metabolism. The results of the determination of the content of glucose, insulin, ketoses, ammonia, urea and amino acids in the blood are announced, the results of glucose elimination contained in the intravenous glucose tolerance test are depicted as tolerance curves. As a result, there was to be seen, arranged according to animal categories, a ranking of the glucose elimination rate of castrated pigs greater than sows greater than boars. The same sequence was observed with regard to insulin secretion, which corresponds to the blood sugar level. On the whole, the results of the blood tests do not show critical states of the metabolism hat can lead to an influentiation of energy metabolism. PMID- 2546516 TI - Use of synthetic oligonucleotide probes to detect rhinovirus RNA. AB - Current methods of detecting a human rhinovirus (HRV) infection are either based on isolation of virus in appropriate susceptible cell lines, which is time consuming and requires considerable expertise, or are dependent on knowing the serotype. The existence of over 100 immunologically distinct serotypes makes serotype specific assays, such as ELISA, unsuitable for general diagnostic assays. In this study a general rhinovirus assay is described which utilises synthetic oligonucleotides as probes in a filter hybridization assay. The probes are designed to bind to short but highly conserved regions of the rhinovirus genome. Indeed, the probes successfully detected all 57 rhinovirus serotypes tested. Furthermore, the test was used to demonstrate rhinovirus infection in clinical samples from 57 volunteers, inoculated with HRV, collected on six consecutive days. Clinical samples were taken prior to inoculation and on days 2 7 after inoculation. The filter hybridization assay gave results comparable to virus culture on days 2 and 3 post-inoculation, but was more sensitive on subsequent days. PMID- 2546515 TI - Sequence of the coding regions from the 3.0 kb and 3.9 kb mRNA. Subgenomic species from a virulent isolate of transmissible gastroenteritis virus. AB - Subgenomic mRNA from a virulent isolate of porcine transmissible gastroenteritis virus (TGEV) was used to produce cDNA clones covering the genome region from the 3' end of the peplomer gene to the start of the integral membrane protein gene. The nucleotide sequence of this area was determined using clone pTG11 and a previously reported cDNA clone pTG22. Three open reading frames (ORFs) were identified encoding putative polypeptides of relative molecular masses (Mr) 6,600, 27,600, and 9,200. The sequence encoding the Mr 9,200 polypeptide was found to be present on the "unique" 5' region of the 3.0 kb mRNA species whereas the other two ORFs mapped on the 3.9 kb mRNA species. Differences between the ORFs from this strain of TGEV and those from a previously reported avirulent strain of TGEV were compared. PMID- 2546517 TI - An ELISA blocking test using a peroxidase-labelled anti-HN monoclonal antibody for the specific titration of antibodies to avian paramyxovirus type 1 (PMV1). AB - This report describes an ELISA blocking test using a peroxidase-labelled monoclonal antibody which binds to the HN protein of Newcastle disease (NDV). This test allows specific detection of type 1 avian paramyxovirus (PMV1) antibodies but does not detect other avian paramyxovirus (PMV2-9) antibodies recognized by the usual serological NDV tests (HI, Orgenics, and Agritech ELISA tests). Furthermore, swollen head syndrome and influenza antibodies were also not detected. ELISA blocking and HI titers of sera collected from SPF chickens immunized with 18 different PMV1 strains (including pigeon isolates) were the same; the correlation between ELISA blocking and HI titers was highly significant (P less than 0.001). In comparison with ELISA tests available commercially at the present time, the ELISA blocking test can be performed more quickly and is applicable without modification to sera from different species of fowls. For this reason, the test appears to be useful for determining the immunity and sanitary status of fowls. When recombinant or deleted vaccines become available, the test should make it possible to demonstrate with confidence any infection of fowls by wild type PMV1. PMID- 2546518 TI - Characterization of a new polyomavirus (Polyomavirus papionis-2) isolated from baboon kidney cell cultures. AB - Viruses with papovavirus morphology were seen in fluids from baboon kidney cell cultures on three separate occasions (isolates A, B, and C). The size of the virions, 47.9 nm, placed the virus in the polyomavirus genus. It grew well in baboon kidney and Vero cells and less well in human embryo lung (HEL) fibroblasts. The virus could not be identified as the previously described baboon polyomavirus, SA 12, or as any of the other known primate polyomaviruses BK, JC or SV 40, the non-primate viruses mouse polyoma, K, rabbit kidney vacuolating virus (RKV) or bovine polyomavirus (FRKV) by immunofluorescence, immune electron microscopy or hemagglutination inhibition (HI) tests. A rabbit antiserum to the new virus (isolate A) reacted only with the three isolates and not with the other primate polyomaviruses studied. Thirteen percent of 118 wild-caught baboons (Papio anubis) had HI antibody to the new polyomavirus and 21 percent were seropositive for SA 12; only two baboons had antibody to both viruses. These results suggest that in baboons there are two antigenically distinct polyomaviruses which circulate independently. The two viruses may also be distinguished by their hemagglutinating properties: SA 12 agglutinated erythrocytes from a wider range of species but only the newly recognized polyomavirus agglutinated baboon erythrocytes. We propose that the two baboon viruses, SA 12 and the new virus, should be named Polyomavirus papionis-1 and Polyomavirus papionis-2 respectively. PMID- 2546519 TI - Spread of herpes simplex virus type-1 (Miyama +GC strain) to the central nervous system after intraperitoneal inoculation: the role of the myenteric plexus of the gut. AB - The pathways taken by the HSV-1 virus after intraperitoneal (i.p.) inoculation were studied in 5-week old male C3H/HeN mice injected with 1 x 10(4) PFU (100 LD50) or 5 x 10(5) PFU (5000 LD50) of HSV-1 (Miyama +GC strain). At the higher dosage (5 x 10(5) PFU), HSV-1 began replicating in the adrenal from the first day, then in the gut and thoracic portion of the spinal cord by the third day, and in the brainstem by the fourth day, as shown by the titers of the virus in these organs. By immunoperoxidase staining HSV-1 was localized in a necrotic area of the adrenal, the myenteric plexus of the gut, the intermediolateral columns of the thoracic cord, and the vagus nerve nuclei of the medulla oblongata. In the low dose mice (1 x 10(4) PFU), HSV-1 was not isolated from the adrenal or thoracic segment of the spinal cord from the time of inoculation until the time of death. It was, however, isolated from the gut on days 4-6 and from the brainstem by day 5. HSV-1 was never isolated from the blood of either group at any time. The localizations of viral replication suggest that in the mice inoculated with 1 x 10(4) PFU, HSV-1 spreads to the brainstem via the vagal nerves after replication in the myenteric plexus of the gut. In the mice given the higher dose, localizations suggest not only the above route, but also that the virus spread to the intermediolateral columns of the spinal cord after replicating in the adrenal. PMID- 2546521 TI - [Study of development of autoantibody to beta-adrenergic receptors in asthmatics]. AB - An (125I) iodohydroxybenzyl pindolol (125IHYP) binding inhibition assay was performed. Various dilutions (1:5-1:500) of sera from asthmatics and controls were incubated with canine lung membranes for 60 minutes at 30 degrees C. 125IHYP was added to the membranes for 30 minutes at room temperature in the presence and absence of 10 microM 1-propranolol, and the samples were washed through a Gelman (Type A-E) glass fiber filter using a washing buffer. Radio activity was measured with Aloka gamma counter. In the presence of various serum dilutions from asthmatics, 125IHYP specific bindings of 0.16 fmol to 4.38 fmol were measured. 125IHYP binding was inhibited in a dose-related and nonspecific manner. Serum, albumin, L-histidine and L-cysteine also inhibited 125IHYP specific binding to beta-receptors. Percentages of inhibition of serum from asthmatics on 125IHYP specific finding to beta-receptors were -17.6% to +9.3%, which were compared with identical dilutions of control serum. There was no significant difference in 125IHYP binding inhibition assay between asthmatics and controls. From these results, development of autoantibody to beta-adrenergic receptors could not be detected in this study. PMID- 2546522 TI - [Effect of phorbol myristate acetate (PMA) on diphosphoinositide (DPI) synthesis in rat mast cell granules]. AB - Rat mast cell granules were obtained by homogenization of highly purified rat mast cells and isolated in a Percoll gradient. DPI synthesis in rat mast cell granules was assayed by measuring the incorporation of 32P from [gamma 32P] ATP into DPI in the absence of exogenous phosphatidylinositol (PI). Lipids were isolated with methanol/chloroform/HC1 and were separated by thin-layer chromatography on oxalic acid impregnated silica gel plates. DPI areas were identified by staining with iodine, scraped and measured for 32P radioactivity. The addition of PMA to the granules caused an increase of DPI synthesis, which can be catalysed by PI kinase. Neither an inactive phorbol ester, 4-alpha-phorbol 12, 13-didecanoate, nor dimethyl sulfoxide (DMSO) used as a solvent for PMA had any effect. The effect of PMA in the DPI synthesis was dose-dependent and maximal effects were observed at 10-100 ng/ml. Dose-response curves of the effects of PMA in DPI synthesis in the granules corresponded to those of other biochemical effects of PMA in rat mast cells, such as mediator release mediated through the activation of protein kinase C. These results suggest that PMA may directly affect PI kinase or indirectly regulate its activity in rat mast cell granules. PMID- 2546523 TI - Ocular varicella-zoster virus infection in the guinea pig. A new in vivo model. AB - Corneal intrastromal inoculation of guinea pigs with approximately 10(4) plaque forming units of live, adapted varicella-zoster virus (VZV) resulted in reproducible, acute, superficial corneal disease in all animals. The culture positive VZV ocular infection progressed to involve 30% to 40% of the corneal surface in a diffuse punctate keratitis and 10% to 15% of this surface with microdendrites, characteristic of VZV-induced ocular disease. Retrograde dissemination of VZV to the trigeminal ganglia, midbrain, cerebellum, and superior cervical ganglia was demonstrated by whole-cell coculture VZV recovery. Central nervous system VZV dissemination, manifested by transient neurologic symptoms and pneumonitis, was evident in 60% of the animals. Varicella-zoster virus spread to the trigeminal ganglion during acute and early-latent infection was evident by electron microscopy. PMID- 2546520 TI - Persistence of rat parvovirus in athymic rats. AB - Euthymic (SD or outbred rnu/+) and athymic (rnu/rnu) rats were inoculated oronasally or intraperitoneally with the RV-Y strain of rat virus when they were 2 days or 4 weeks old. Clinical signs of infection in athymic infants were similar to those in euthymic infants, but significantly more athymic infants died. Some infants developed anemia and thrombocytopenia. After inoculation of infants. RV-Y was detected in surviving euthymic rats for 7 weeks and in surviving athymic rats for at least 10 weeks. After oronasal inoculation of 4 week-old rats no clinical illness was observed. RV-Y persisted less than 6 weeks in juvenile euthymic rats but at least 12 weeks in athymic juvenile rats. Intraperitoneal inoculation of juveniles resulted in infection for at least 6 weeks. The antibody response of athymic rats to RV-Y was significantly reduced compared to that of euthymic rats. These studies indicate that T cell deficiency increases the severity and duration of RV infection and imply that T cells are required for the full expression of resistance to RV infection. They also suggest that RV-Y induced anemia could serve as a model for human parvovirus-induced anemia. PMID- 2546524 TI - Management of chronic blepharitis. PMID- 2546525 TI - Ocular manifestations in pediatric patients with acquired immunodeficiency syndrome. AB - Forty pediatric patients seropositive for human immunodeficiency virus antibody and conforming to Centers for Disease Control, Atlanta, Ga, case definition of acquired immunodeficiency syndrome underwent ophthalmic examinations to evaluate prospectively the incidence, type, and natural history of ocular involvement in pediatric acquired immunodeficiency syndrome. A total of 87 examinations were performed on the patient population throughout the course of the study. Twenty percent had ocular findings, including two cases of cytomegalovirus retinitis, one case of isolated retinal cotton-wool spots, one case of toxoplasmosis retinochoroiditis, and three cases of external infections of adnexal structures. One patient had unusual peripheral retinal findings. The incidence of ocular manifestations in pediatric acquired immunodeficiency syndrome is considerably less than reported in several adult series. However, we recommend ophthalmic screening in all pediatric patients with acquired immunodeficiency syndrome with encephalopathy or disseminated opportunistic infections, or when symptoms suggest ophthalmic involvement. PMID- 2546527 TI - Bacteria and gallstone nucleation. AB - This preliminary study reports for the first time that there might be a possible association between bacteria and the aetiology of some cholesterol calculi. The gall-bladder biles from 225 cholecystectomy patients underwent bacteriological and microscopic study. Cholesterol calculi from 13 patients (10.2%) were observed to be associated with gall-bladder bile profusely infected with at least one bacterial species that was shown to possess beta-glucuronidase activity, an enzyme that is thought to promote calcium bilirubinate precipitation in bile. Concomitantly, the associated gall-bladder bile was 'high' in calcium bilirubinate precipitation and the precipitate was also detected in the centre of the gallstones. Moreover, in approximately half of these patients (six of 13), the cholesterol gallstones' nuclear areas also contained calcium palmitate, which is also thought to be due to bacterial activity. The results also support the hypothesis that bacteria with active beta-glucuronidase (for example, Escherichia coli) can significantly influence the aetiology of brown pigment gallstones. In contrast, bacteria were observed to have no role in black pigment gallstone formation, as their associated gall-bladder biles were always observed to be sterile. PMID- 2546526 TI - Beta-very low density lipoprotein uptake in cultured fibroblasts and smooth muscle cells from Watanabe heritable hyperlipidemic rabbits. AB - Watanabe Heritable Hyperlipidemic (WHHL) rabbits are an important animal model used to study the effects of defective low density lipoprotein (LDL) receptors on lipoprotein metabolism. In the present study, the receptor-mediated catabolism of apolipoprotein (apo) E-containing lipoproteins was investigated in fibroblasts and smooth muscle cells cultured from WHHL rabbits. Fibroblasts from WHHL rabbits bound little, if any, human LDL, an apo B-containing lipoprotein, in 4 degrees C equilibrium binding experiments. In contrast, canine beta-very low density lipoproteins (beta-VLDL), which contain apo E, bound to these cells with an affinity similar to that for fibroblasts of normal New Zealand White rabbits (Kd approximately 0.6 micrograms/ml), although the maximal binding was only approximately 9% of that of New Zealand White cells. At 37 degrees C, beta-VLDL were degraded efficiently by WHHL fibroblasts, that is, at rates approximately half of that for New Zealand White fibroblasts. The degradation of beta-VLDL by WHHL fibroblasts was blocked by reductive methylation of the lipoproteins and was inhibited by the anti-apo E monoclonal antibody, 1D7. Unlike human LDL, canine beta-VLDL induced 14C-oleate incorporation into cholesteryl esters in WHHL fibroblasts and smooth muscle cells. The uptake of beta-VLDL by WHHL cells was mediated by the LDL receptor, since preincubation of the fibroblasts with cholesterol plus 25-hydroxycholesterol inhibited beta-VLDL degradation, and specific antibodies to rabbit LDL receptors abolished the degradation of beta VLDL in WHHL fibroblasts. These results indicate that the altered LDL receptors expressed on cultured WHHL rabbit cells are sufficient for apo E-dependent, receptor-mediated uptake of lipoproteins. Thus, the altered LDL receptors could contribute to in vivo metabolism of apo E-containing lipoproteins despite their lack of LDL binding activity. PMID- 2546528 TI - Vertical transmission of reticuloendotheliosis virus in breeder turkeys. AB - Epizootiological studies were conducted on a commercial turkey breeder flock naturally infected with nondefective reticuloendotheliosis (RE) virus. Although RE virus was isolated from 27 (46%) of the 59 hens studied, only 4 of the 45 hens tested transmitted RE virus to progeny during a 6-week observation period and the overall transmission rate was 1.8%. The transmitter hens were of two types: three hens were consistently viremic and antigenemic and lacked antibody; one hen was viremic but lacked detectable viral antigen and possessed antibody. Toms appeared to play no role in vertical transmission of the infection. Of several tests evaluated for detection of transmitter hens, the direct enzyme-linked immunosorbent assay on albumen was probably best, since it detected three of four transmitter hens, detected relatively few nontransmitter hens, and had the best consistency of any test. No significant differences in hatchability were found between eggs from viremic and non-viremic hens. These findings can be utilized in the development of programs for eradication of RE virus from turkey breeding flocks. PMID- 2546529 TI - Pathogenesis of Marek's disease virus-induced local lesions. 1. Lesion characterization and cell line establishment. AB - Subcutaneous (wing-web) or intramuscular inoculation of chickens with allogeneic normal or Marek's disease virus (MDV)-infected chicken kidney cells induced local lesions visible by 3-4 days postinoculation (PI). Lesions were slightly larger (P less than 0.05) in infected than uninfected chickens 5 and 8 days PI. They persisted and grew past 9 days PI only when infected. Infiltrating lymphocytes in infected and uninfected early lesions were similar; they included B-cells and also T-cells with and without Ia antigen. Up to 42% of lymphocytes from infected or uninfected lesions had the surface antigen MATSA. At 3 to 6 days PI, infected lesions contained lymphocytes with viral internal antigen, especially in Ia bearing cells and MATSA-bearing cells, but thereafter infection was latent. Cells harvested daily from local lesions induced with allogeneic MDV-infected cells were cultured; MD tumor cell lines were established from lesions as early as 4 days PI, with a total success rate of about 50% thereafter. Either transformed tumor cells were already present during the early cytolytic infection period or else appropriate target cells were present that became infected in vivo and/or in vitro and then became transformed in vitro. PMID- 2546530 TI - Restriction endonuclease analysis of infectious laryngotracheitis viruses: comparison of modified-live vaccine viruses and North Carolina field isolates. AB - Six modified-live (ML) infectious laryngotracheitis (ILT) vaccine viruses, three reference strains, and 18 field isolates were compared by restriction endonuclease analysis of their DNA. Viral DNA digestion patterns were established for vaccine viruses using restriction endonucleases PstI, BamHI, KpnI, and HindIII. Using these enzymes, five of six ML vaccine viruses had identical restriction endonuclease cleavage patterns. Vaccine viruses had distinct patterns compared with ILT virus reference strains Illinois-N71851, Cover, and NVSL. Restriction endonuclease cleavage patterns of 18 field isolates of ILT virus, obtained from ILT outbreaks in North Carolina, were indistinguishable from vaccine viruses. These results suggest a possible role of vaccine or vaccine-like viruses in recent ILT outbreaks. PMID- 2546532 TI - A proposed protocol for clinical trials of boron neutron capture therapy in glioblastoma multiforme. PMID- 2546531 TI - Pathologic changes in chickens caused by intravenous inoculation with fowlpox virus. AB - Twenty chickens were inoculated intravenously with fowlpox (FP) virus, and clinical and pathological examinations were carried out chronologically. Upon gross examination, miliary nodules scattered in the kidneys were observed from 10 to 18 days postinoculation (PI), as were papules on the skin and diphtheritic lesions on the mucous membrane of the upper respiratory tract. Microscopically, characteristic FP lesions, composed of swelling and proliferation of cells with formation of Bollinger bodies, were observed in the epithelial cells of renal tubules from 4 to 14 days PI and in the epithelial reticular cells of the thymic medulla from 4 to 10 days PI, as well as in the skin and mucous membrane. Immunofluorescent and electron microscopic observations confirmed the presence of viral antigen and virus particles in the characteristic lesions of FP. PMID- 2546533 TI - Computerized axial tomographic and magnetic resonance imaging scan follow-up of two patients after boron neutron capture therapy for glioblastoma multiforme. PMID- 2546534 TI - Working group on tumor compound and compound delivery system. PMID- 2546535 TI - Fractionation in boron neutron capture therapy. PMID- 2546536 TI - [Determination of the activity of superoxide dismutase in terms of rea substrate conversion]. AB - On the basis of photochemical generation of O2.- and the tetrazolium/formazan system as indicating reaction an assay for SOD was developed which allows the determination of initial rate velocities of the enzyme. This was achieved by a special apparatus yielding the continuous registration of product formation simultaneously with the photochemical generation of O2.-. The investigation of the indicating reaction revealed that the reduction of nitro blue tetrazolium chloride to formazan is stoichiometrical with respect to riboflavin/methionine generated O2.- above pH 9. On this basis, an activity unit of SOD can be defined as real substrate consumption. One unit of the enzyme causes a decrease in the formation of formazan of 1 mumol/min under defined conditions. PMID- 2546537 TI - Purification of cytochrome P-450 from n-hexadecane-grown Acinetobacter calcoaceticus. AB - A convenient procedure for the purification of cytochrome P-450 from an n-alkane assimilating strain of the bacterial species Acinetobacter calcoaceticus has been elaborated. The cytochrome P-450 of n-hexadecane-grown cells was found to be distributed in cell-free extracts among particulate and soluble subcellular fractions. For isolation, cytochrome P-450 was extracted from particulate fractions by addition of Triton X-100 to the buffer. Subsequently, purification to an apparently homogeneous state was achieved by chromatography on DEAE cellulose, Sepharose CL-6B and hydroxylapatite cellulose. A Mr of 52,000, an isoelectric point of 4.7 and the amino acid composition were determined. The Soret bands of absolute absorption spectra showed maxima at 417 nm for the oxidized form, at 408 nm for the reduced form and at 448 nm for the carbon monoxide compound of the reduced cytochrome. Difference spectra with octylamine showed maxima at 432 nm and minima at 410 nm indicating the predominance of the low-spin state. Conversion to the high-spin state could not be obtained. The isolated cytochrome P-450 was stable in the presence of Triton X-100 under neutral pH conditions. Removal of the detergent or change of the pH to values higher than 8.0 or lower than 6.0 resulted in the destruction of the cytochrome P 450. PMID- 2546538 TI - Evidence for cholinergic participation in frontal cortical potentials evoked by single impulse stimulation in the locus coeruleus area of freely moving rats. AB - Single impulse stimulation in the area of locus coeruleus (LC) in freely moving rats evokes bilateral symmetrical potentials (EP) in the frontal cortex. Threshold intensity for this potential was higher when the stimulus electrode was in the LC than in the cholinergic dorsal parabrachial nucleus lateral of LC. Amplitudes and peak times of this EP were constant during relaxed wakefulness, exploratory behavior and unaffected by beta-blockade (propranolol, 2.5 mg/kg i.p.). Their amplitudes were strongly decreased and the peak times of later negativity (N50) were prolonged during slow wave sleep and by muscarinic blockade with scopolamine (0.5 mg/kg i.p.) which induced active behavior and synchronized EEG. These data suggest a direct participation of cholinergic structures in cortical potential evoked by electrical LC stimulation. Relations between stimulus effect and behavior regulating systems are discussed. PMID- 2546540 TI - Patch clamp analysis of recovery of sodium channels from inactivation in mammalian myocardium in terms of a Markovian state model. AB - The kinetics of the recovery of Na channels from inactivation were investigated in ventricular cardiocytes of adult mice by a patch clamp technique. Single channel analysis showed an unaltered slope conductance of unitary currents during the recovery process to be 18.5 pS. The mean open time tau 0 and its monoexponential distribution remained also unchanged. The recovery time course of the apparent probability for a channel not to open coincided perfectly with the recovery time course determined from peak values of macroscopic currents which were recorded from cell-attached patches with up to 80 channels by averaging between 19 and 38 sweeps. Conclusively, the recovery was quantified by the time course of re-accumulation of the channels in a resting state. The initial recovery time course was sigmoidal. A serial Markov state model with three inactivated states in addition to the resting state was most appropriate for its description. The rate constants were assumed to be equal direction. A highly voltage-sensitive reaction towards the last inactivated state and a nearly voltage-independent reaction towards the resting state were obtained. Slow inactivation components were analyzed both during the recovery process (-100 mV ... -130 mV) after long preceding inactivation and at maintained depolarization ( 50 mV ... -20 mV). For description, the model was extended by two further, serially coupled inactivated states. All four rate constants describing slow inactivation in the range of hundreds of milliseconds to seconds proved to be voltage-independent. Taken together, the results suggested the absorbing inactivated state I to be composed of five serially coupled inactivated states with only two transitions being steeply voltage-dependent. PMID- 2546539 TI - Modulatory effect of locus coeruleus stimulation on visually evoked potentials in freely moving rats. AB - Interactions between light-flash stimulation and electrical single impulse stimulation of locus coeruleus (LC) were investigated in 9 freely moving rats with chronically implanted electrodes during different behavioral states. The interaction of both showed different immediate and long-lasting effects. Simultaneous visual and electrical stimulation caused a clear decrease of trigger rate of photically evoked after-discharges from 60% to 15% and a reduction of the early primary component of visually evoked potentials (VEP) in area 17 compared with the values without LC stimulation during relaxed wakefulness (RW). On the contrary, the simultaneous LC stimulation was ineffective during exploratory behavioral states (E). At 100 ms stimulus interval and more there was no detectable LC stimulus effect on VEP components during RW, whereas the LC stimulation under beta-blockade (2.5 mg/kg propranolol i.p.) during RW-like behavior clearly reduced the early VEP component to 75% compared with values without LC stimulation. On the other hand, LC stimulation facilitated mainly the primary VEP component to about 160% during E compared with the flash response alone. This effect was observed from 100 to 300 ms after LC stimulus and was unaffected by muscarinic blockade (0.5 mg/kg scopolamine i.p.). We conclude from these data that the short-time effects are comparable with the influence of spontaneous behavioral activation and probably result from a costimulation of cholinergic neurons around LC, whereas the long-lasting effects may be attributed to the influence of noradrenergic LC cells. The noradrenergic-dependent facilitation of early VEP component counteracts the cholinergically-induced reduction. PMID- 2546541 TI - Effects of intraperitoneally applied D-galactosamine on uridine and cytidine plasma content and brain activity of uridine kinase in the rat. AB - As determined by the HPLC technique, injection of 1.85 mmoles/kg D-galactosamine caused strong alterations in the plasma level of cytidime. Thus, 2 h and 4 h after application of the amino sugar only 54% and 61%, respectively, of the normal values were measured, while an increase to 135% of controls was observed 24 h later. In contrast, intraperitoneal application of D-galactosamine had no influence on the plasma level of uridine. The activity of the enzyme, uridine kinase, in the hippocampal area of the rat brain was increased 2 h and 4 h after D-galactosamine treatment. The results are discussed in the light of behavioural findings in which intraperitoneally applied D-galactosamine at this dosage improves the retention performance of an acquired behaviour in the rat. PMID- 2546542 TI - Identification and characterization of endothelin binding sites in rat renal papillary and glomerular membranes. AB - The present study was designed to identify and characterize specific endothelin binding sites in membranes of rat renal papillae and glomeruli which appear to be target tissues for this new peptide hormone. Saturation binding studies indicate that the sites have a high and uniform affinity. The dissociation constants averaged 662 +/- 151 and 1309 +/- 123 pM and the receptor densities 7666 +/- 920 and 5831 +/- 348 fmol/mg protein for papillary and glomerular membranes, respectively. Endothelin 1, endothelin 3 and sarafotoxin all inhibited [125I] endothelin binding with IC50's in the 100-300 pM range, whereas unrelated peptides, namely angiotensin II, atrial natriuretic peptide, and platelet-derived growth factor failed to compete for [125I]-endothelin binding. Deletion of the carboxyterminal tryptophan in endothelin 1 reduced its affinity for glomerular binding sites by 2 orders of magnitude. Specific endothelin binding to these membranes was maximal at pH 4 and was markedly inhibited as the pH was raised above 8. When [125I]-endothelin is covalently linked to glomerular membrane binding sites, SDS-PAGE of these solubilized membranes followed by autoradiography reveals a predominant specifically labeled band of 45 kDa. Whether this band represents a subunit of the endothelin receptor(s), the receptor proper, or an intracellular endothelin binding protein remains to be determined. PMID- 2546544 TI - Phosphorylation of calf uterine progesterone receptor by cAMP-dependent protein kinase. AB - We have examined the potential for using calf uterine progesterone receptor (PR) as a substrate for phosphorylation by cAMP-dependent protein kinase (cAMP-PK), PR was found to interact with anti-PR monoclonal antibody alpha PR6 (Sullivan et al., 1986), which was to immunopurify the receptor. Protein staining of the purified preparation revealed the presence of two major bands corresponding to 114 kDa and 90 kDa peptides; only 114 kDa peptide could be photoaffinity-labeled with R5020. The 90 kDa peptide co-migrated with 90 kDa heat shock protein (hsp 90) precipitated by anti-hsp-90 monoclonal antibody AC88 (Riehl et al., 1985). Incubation of the immunopurified protein-A-Sepharose-adsorbed PR with the catalytic subunit of cAMP-PK in the presence of gamma-[32P]ATP and divalent cations resulted in a Mg++-dependent incorporation of 32P-radioactivity into both the 114 kDa and the hsp-90 peptides. Small 32P-incorporation was also seen in the 114 kDa peptide in the presence of Mn++. A 60 degrees C preincubation of immunopurified PR increased the extent of phosphorylation of the hsp-90 peptide. A pretreatment with alkaline phosphatase reduced the ability of PR to act as a substrate while the steroid occupancy of PR appeared to enhance the phosphorylation of the 114 kDa peptide. The differential cation requirement for the phosphorylation of 114 kDa and hsp-90 peptides and a selective hormone dependent increase in the phosphorylation of the 114 kDa peptide suggest a possible role of phosphorylation in mediating progesterone action in the calf uterus. PMID- 2546543 TI - Effects of H-7 are not exclusively mediated through protein kinase C or the cyclic nucleotide-dependent kinases. AB - Culturing murine T cell tumor lines in the presence of the protein kinase inhibitor H-7 for 4 days led to their dependence on H-7 for maximal constitutive proliferation. Withdrawal of H-7 from H-7-conditioned cells led to inhibition of proliferation and cell death. The mechanism underlying this H-7 dependence does not appear to be related to clonal selection or to effects on protein kinase C or the cyclic nucleotide-dependent kinases. This suggests that all the effects of the widely used H-7 may not be completely understood, and that H-7 may be useful in the dissection of the complex patterns of growth regulation in T cell malignancies. PMID- 2546545 TI - Rate enhancement of the electron transfer the adrenodoxin-adrenodoxin reductase system by dicarboxylic acids. AB - The rate of electron transport in the cytochrome P-450 system in adrenocortical mitochondria was studied with purified adrenodoxin reductase, adrenodoxin and cytochrome c. Oxaloacetate enhanced the rate at concentrations of less than 1 mM; malate, succinate and fumarate enhanced the rate to a lesser extent; and pyruvate and alpha-ketoglutarate had no appreciable effect. The rate enhancement was observed when the reagents were preincubated with adrenodoxin, but not with adrenodoxin reductase. Rate enhancement was also evident when the rate limiting step was at adrenodoxin in the electron transport system. PMID- 2546546 TI - Characterization of a topoisomerase-like activity at specific hypersensitive sites in the Drosophila histone gene cluster. AB - It is well known that treatment of DNA-topoisomerase complexes with SDS induces cleavage of the DNA by trapping a reactive intermediate in which the topoisomerase is covalently linked to the terminal phosphates of the cut DNA. I have used this technique to examine potential topoisomerase binding sites in the histone gene chromatin of Drosophila Kc cells. Treatment of Kc nuclei with SDS induces Mg++-dependent DNA cleavage near the borders of two nuclease hypersensitive sites located 5' and 3' of histone H4. It is likely that the SDS induced cleavage at these hypersensitive sites is due to a topoisomerase because protein becomes tightly bound to the ends of the cleaved DNA fragments. Preliminary experiments suggest that a type II topoisomerase may be responsible for the cleavage. PMID- 2546547 TI - Monitoring of the activation of receptor-operated calcium channels in human platelets. AB - Activation of receptor-operated calcium channels has been monitored by measurements of the quenching of the fluorescence of intracellularly trapped fura 2 by Mn entering from the extracellular medium. Release of calcium from intracellular stores was followed simultaneously by measurements of the ratio of the fluorescence excited at 340 and 380 nm. Thrombin, ADP, platelet-activating factor (PAF) and collagen, all produced both release of calcium from the intracellular stores and uptake of Mn from the extracellular medium. The uptake of Mn, but not the increase of (Ca2+)i, was blocked by nickel. These results suggest the existence of plasma membrane calcium channels which can be activated by the different agonists tested here. The activation of calcium channels was very fast and transient with ADP and PAF, fast and maintained with thrombin, and delayed with collagen. PMID- 2546548 TI - Guanine nucleotides stimulate hydrolysis of phosphatidyl inositol bis phosphate in human myelin membranes. AB - Phosphodiesterase activity was stimulated in myelin membranes in the presence of guanine nucleotide analogues. This activity was reduced in myelin membranes which had been adenosine diphosphate ribosylated in the presence of cholera toxin which ADP-ribosylated three proteins of Mr 46,000, 43,000 and 18,500. Aluminum fluoride treatment of myelin had the same stimulatory effects on phosphodiesterase activity as did the guanine nucleotides. PMID- 2546549 TI - Molecular changes associated with induction of cell death in a human T-cell leukaemia line: putative nucleases identified as histones. AB - Following treatment of the human T-cell leukaemia line, CEM-C7, with the glucocorticoid, dexamethasone, a rapid decrease in viability occurred after 40 h which coincided with fragmentation of DNA in these cells. A similar pattern of DNA fragmentation was observed when these cells were gamma-irradiated or treated with cycloheximide. Distinct morphological changes occurred after treatment, indicating a form of cell death, regulated from within, termed apoptosis. A set of nuclear proteins ranging in size from 10-18 kDa appeared by 40 h following treatment with dexamethasone. Treatment of cells with gamma-irradiation or cycloheximide also produced the same protein pattern. This set of proteins, and a doublet approximately 55 kDa in size, had apparent nuclease activity which was not observed in untreated cells. However, protein microsequencing of these bands in the 10-18 kDa region revealed that they were histone proteins. These results cast doubt on a recent report which provided evidence that these proteins were induced nucleases. PMID- 2546551 TI - Extracellular ATP triggers superoxide production in human neutrophils. AB - The effects of ATP on the concentration of cytosolic calcium [( Ca2+]i) were examined with respect to early events associated with activation of the superoxide (O2-)-generating system in human neutrophils. Addition of ATP to cytochalasin B-treated neutrophils resulted in two sequential increase in [Ca2+]i: an initial phase presumably related to the mobilization of Ca2+ from intracellular stores and a second phase dependent upon the presence of extracellular Ca2+. The second phase was associated with an increase in the rate of O2- production, which also required the presence of extracellular Ca2+. The results suggest that increased Ca2+ influx may act to trigger a cascade of Ca2+ sensitive events, leading to stimulated O2-production. PMID- 2546550 TI - Dissociation of alpha-adrenergic and cholinergic stimulated inositol trisphosphate-dependent calcium mobilization at the "post-receptor" level. AB - Carbachol stimulated inositol trisphosphate (IP3) production and subsequent calcium mobilization in parotid cells are almost completely inhibited by neomycin. In contrast epinephrine stimulated IP3 production and calcium mobilization are much less sensitive to such inhibition. Since neomycin exerts its effects primarily at the level of inositol phosphate production and action, cholinergic and alpha adrenergic stimulation of IP3 dependent calcium mobilization may proceed through different "post-receptor" signal transduction mechanisms in parotid cells. PMID- 2546552 TI - Interaction of synthetic sarafotoxin with rat vascular endothelin receptors. AB - The effects of synthetic analogs of sarafotoxin (STX) S6b, a snake venom peptide with a high sequence homology to the endothelium-derived vasoconstrictor endothelin (ET), on ET receptor binding activity and cytosolic free Ca2+ concentration [( Ca2+]i) were studied in cultured rat vascular smooth muscle cells. Binding studies revealed that [Cys1-15, Cys3-11] STX competed with 125I-ET for the binding to its vascular receptors with lower affinity than that of ET, but was far more effective than [Cys1-11, Cys3-15]STX in inhibiting the binding. [Cys1-15, Cys3-11]STX had a less potent effect on increasing [Ca2+]i than ET, whereas [Cys1-11, Cys3-15]STX was inactive. These data suggest that there may exist heterogenous subpopulations of the vascular ET/STX receptors, and that the proper double cyclic structure of STX is essential for interacting with its putative receptors to induce the [Ca2+]i response. PMID- 2546553 TI - Placental estrogen synthetase (aromatase): evidence for phosphatase-dependent inactivation. AB - The acute regulation of estrogen synthetase (aromatase), the cytochrome P450 enzyme system responsible for estrogen production, is not well explored. We report here that aromatase, but not NADPH-cytochrome c (P450) reductase, activity from human term placental microsomes decreased when incubated in phosphate-free buffer at 37 degrees C. Aromatase activity was stabilized by phosphate buffer or by the phosphatase inhibitors tartaric acid or EDTA, but not NaF, in phosphate free buffer. Alkaline phosphatase also inhibited aromatase in phosphate-free buffer relative to phosphate buffer, but the inactivation appears to be due primarily to proteolytic solubilization of NADPH-cytochrome c reductase from the microsomes by proteases within the alkaline phosphatase preparation. Based on these data, we suggest that the cytochrome P450 component of aromatase may be regulated acutely by phosphorylation-dependent processes. PMID- 2546554 TI - Evidence for two platelet activating factor receptors on eosinophils: dissociation between PAF-induced intracellular calcium mobilization degranulation and superoxides anion generation in eosinophils. AB - We have compared platelet activating factor (PAF)-induced eosinophil peroxidase (EPO) release and intracellular calcium mobilization with superoxide anion (.O2-) generation from guinea pig eosinophils. EPO release and Ca2+ mobilization occurred at lower concentrations of PAF (EC50 values of 1.3 nM and 11.5 nM, respectively) while .O2- production was observed at higher concentrations (EC50 of 31.7 microM). Receptor characterization with the competitive PAF antagonist, WEB 2086, gave pA2 values of 8.5 and 8.3 for EPO enzyme release and rise in [Ca2+]i, respectively, and 5.8 for the .O2- production. In addition, PAF-induced degranulation and elevation of [Ca2+]i were dependent on extracellular Ca2+ whereas PAF-stimulated .O2- generation was dependent on the presence of extracellular Mg2+ ions. These results suggest the existence either of two subtypes of the PAF receptor or a single receptor that can exist in one of two affinity states on guinea pig eosinophils. PMID- 2546555 TI - Identification of a disulfide between cysteine 214 and cysteine 277 in the beta subunit of native (Na+ + K+)ATPase. AB - Two peptides, produced during tryptic digestion and thermolytic digestion, respectively, and containing the same intact disulfide from the beta polypeptide of (Na+ + K+)ATPase from Torpedo californica, were isolated and unambiguously identified. The disulfide is between Cysteine 214 and Cysteine 277. PMID- 2546556 TI - Inhibition of ubiquitin-dependent proteolysis by des-Gly-Gly-ubiquitin: implications for the mechanism of polyubiquitin synthesis. AB - Cleavage of the two carboxyl-terminal glycine residues from native ubiquitin yields the proteolysis-incompetent derivative des-Gly-Gly-ubiquitin. We report here that this derivative inhibits the ATP-dependent degradation of casein and is multi-ubiquitinated but not degraded by reticulocyte lysates. Inhibition of proteolysis diminished with increasing concentration of native ubiquitin, but was not reduced by increased casein concentration. Cleavage of the last four residues from ubiquitin yielded a derivative that was a weaker inhibitor of proteolysis and a poorer substrate for ubiquitination. These results suggest that the conjugation of ubiquitin to ubiquitin during polyubiquitin synthesis involves a specific conjugation system that recognizes ubiquitin and some of its derivatives, but not general proteolysis substrates, as ubiquitin acceptors. PMID- 2546557 TI - Roles of tyrosine residue of enkephalin in opiate receptor recognition. AB - A series of analogs of a dimeric peptide of the inactive fragment of enkephalin (H-Tyr-D-Ala-Gly-NH-CH2-)2 (DTRE2) was synthesized by modifying one or both of tyrosine residues. The change of configuration of both tyrosines, from L to D, or the removal of both p-hydroxy groups brought about a decrease in activity, but some of the activity (about 20% of the parent enkephalin dimer's activity) was found to be retained when assayed by using guinea pig ileum and mouse vas deferens. In contrast, the analog lacking amino groups in both tyrosines was completely devoid of activity, indicating the essential importance of the tyrosine amino group in opiate receptor recognition. The activity of analogs with only one amino group was found to be higher than that of the parent enkephalin dimer having both amino groups. A possible interaction to explain this finding was discussed. PMID- 2546559 TI - A biochemical approach to essential hypertension. AB - Using multivariate statistical analysis, an attempt has been made to select hypertensive and normotensive sub-groups of subjects on the basis of certain parameters of their blood serum, such as the inhibition of purified Na,K-ATPase by serum and the content of two proteins with molecular masses of 12 and 15 kDa. An analysis of 20 human beings (10 hypertensive and 10 normotensive individuals) revealed that the best division into sub-groups is achieved only through the use of a combination of these three parameters. PMID- 2546558 TI - Pharmacological intervention in oxidant-induced calcium pump dysfunction of dog heart. AB - Micromolar concentrations of HOCl, an oxidant produced by activated neutrophils, inhibited Ca2+ uptake and Ca2+ATPase of isolated dog heart sarcoplasmic reticulum (SR). DTT antagonized completely the HOCl effect only when it was given within 5 min after the addition of HOCl. When the pharmacological intervention was delayed, the recovery with DTT was not complete, and administration of DTT 30 min after the start of HOCl's reaction with SR resulted in only a small improvement in SR Ca2+ uptake. Although H2O2 and Fe ion-chelate (a free radical-generating procedure) also inhibited Ca2+ uptake and ATPase, the concentrations required were very large. The response of cardiac sarcolemmal and skeletal muscle SR calcium pumps to oxidants was similar to that of the cardiac SR calcium pump. PMID- 2546561 TI - Quantitative dissociation of glucose transport stimulation and insulin receptor tyrosine kinase activation in isolated adipocytes with a covalent insulin dimer (B29,B29'-suberoyl-insulin). AB - The covalent insulin dimer B29,B29'-suberoyl-insulin was investigated for its effects on insulin receptor binding, insulin receptor tyrosine kinase activity and glucose transport in isolated adipose cells. The dimer stimulated glucose transport (initial 3-O-methylglucose uptake rate) to the same extent as insulin did (basal rate, 35 +/- 3 pmol/sec/microliter lipid; insulin, 380 +/- 27; B29,B29'-suberoyl-insulin, 369 +/- 24, means +/- S.E.), although at higher concentrations (EC50 1.94 +/- 0.64 nM versus 0.1 +/- 0.02 with insulin). In contrast, the dimer only partially (23%) mimicked insulin's effect on phosphate incorporation into insulin receptors immunoprecipitated after equilibration of cells with [32P]phosphate. Similarly, insulin receptor tyrosine kinase as assessed by receptor autophosphorylation and phosphorylation of the substrate poly-(Glu/Tyr) was not fully activated by treatment of cells with the insulin dimer (31 and 42% of the effect of insulin, respectively) in concentrations which maximally activate glucose transport and give rise to full insulin receptor occupancy (5 X 10(-7) M). Further, the dimer activated the receptor tyrosine kinase in solubilized purified insulin receptor preparations from adipose cells to only 25% of the effect of insulin (EC50 32.0 +/- 16 versus 1.9 +/- 1.0 nM with insulin) in spite of full receptor occupancy. Binding of the dimer to insulin receptors followed single site binding kinetics, indicating that the derivative is unable to induce negative cooperativity of the insulin receptor. It is concluded that a partial phosphorylation of insulin receptors and a submaximal tyrosine kinase activation are sufficient for full stimulation of glucose transport in the adipocyte. Further, it is suggested that negative cooperativity of the insulin receptor and activation of its tyrosine kinase require a similar conformational change of the receptor protein. PMID- 2546560 TI - Role of ion channels and intraterminal calcium homeostasis in the action of deltamethrin at presynaptic nerve terminals. AB - Using a continuous perfusion system, synaptosomes prepared from rat brain released [3H]norepinephrine in a Ca2+-dependent manner when pulse depolarized by briefly elevating external potassium concentrations. Tetrodotoxin (10(-7) M), a sodium channel blocker, inhibited 48% of this pulsed release, and D595 (10(-5) M), a phenethylamine-type calcium channel blocker, inhibited 21%. In combination, these two specific ion channel antagonists appear to function independently of each other in an additive fashion. Addition of deltamethrin to this preparation resulted in an enhanced release of [3H]norepinephrine which occurred in a biphasic fashion. At 10(-7) M, deltamethrin produced a 42% enhancement in the first or initial peak of [3H]norepinephrine release and a 100% enhancement in the second or tailing peak. Addition of deltamethrin to tetrodotoxin-pretreated synaptosomes resulted in a net 37% enhancement of the initial peak release and a net increase of 277% in the tailing peak. Addition of deltamethrin to D595 pretreated synaptosomes produced no significant effect on enhanced [3H]norepinephrine release from either peak. Since tetrodotoxin is a specific sodium channel blocker, deltamethrin may be enhancing [3H]norepinephrine release by increasing the uptake of Ca2 via other voltage-gated channels (e.g. calcium) or exchange mechanisms in addition to its action at voltage-gated sodium channels. To determine whether deltamethrin may also have an effect on intraterminal Ca2+ homeostasis, external Ca2+ was replaced with Ba2+ and synaptosomes were depolarized with pentylenetetrazole (PTZ). At 10(-5) M, deltamethrin produced a 66% increase in neurotransmitter release over that produced by PTZ alone. An estimated EC50 value of deltamethrin for PTZ-induced release was calculated to be 2.4 x 10(-10) M. PMID- 2546562 TI - Lack of competition between cytochrome c and anthraquinone type drugs for the reductive sites of NADH dehydrogenase. AB - We have shown that (i) the cytochrome c reductase activity of the commercial NADH dehydrogenase does not perturb its ability to catalyse the reduction of various antitumor compounds of the anthracycline class, (ii) the reduction of these compounds by NADH, catalysed by commercial NADH dehydrogenase, correlates with their reduction by NADH catalysed by microsomes. Moreover, our data strongly suggest that two catalytic sites are present, one for cytochrome c and one for quinone type compounds. PMID- 2546563 TI - Inhibition by the LTD4 antagonist, SR2640, of effects of LTD4 on canine polymorphonuclear leukocyte functions. AB - The sulfidopeptide leukotriene LTD4 selectively inhibited directed migration of canine polymorphonuclear leukocytes towards LTB4 (100 nM) with an IC50 of 38 nM. No effect on PAF-acether induced migration was observed. LTD4 did not cause detectable adherence of cells to the Boyden chamber system. Neither LTD4 nor the LTD4/LTE4 receptor antagonist, SR2640, possessed chemokinetic properties. LTD4 induced reversible aggregation of the cells with an EC50 of 36 nM. SR2640 suppressed or abolished LTD4 responses in vitro and following oral administration of the drug. SR2640 had no effect on aggregation induced by LTB4 or PAF-acether. The results can be taken as evidence that canine polymorphonuclear leukocytes possess LTD4 receptors that may be involved in a leukotriene specific micro feedback system between the different cell types involved in inflammatory responses. PMID- 2546564 TI - Induction of aryl hydrocarbon hydroxylase and demonstration of a specific nuclear receptor for 2,3,7,8-tetrachlorodibenzo-p-dioxin in two human hepatoma cell lines. AB - Two established human hepatoma cell lines, Hep3B and HepG2, were examined for aryl hydrocarbon (benzo[a]pyrene) hydroxylase (AHH) induction and for the presence of the murine-equivalent aromatic hydrocarbon (Ah) receptor. Both cell lines demonstrated polycyclic aromatic hydrocarbon (PAH)-induced AHH activity; however, assay conditions for induction were different than those established for the control mouse hepatoma cell line, Hepa c1-9. When cytosols from either cell line were exposed to tritiated 2,3,7,8-tetrachlorodibenzo-p-dioxin [( 3H]TCDD) and analyzed on sucrose gradients with or without prior charcoal treatment, two peaks were observed at positions corresponding to 4-5 S and 8-9 S. The 8-9 S peak was identified as the probable human Ah receptor equivalent since, like the mouse Ah receptor, this peak: (a) was eliminated only by cytochrome P1-450 inducers; (b) was sensitive to protease digestion; and (c) was thermolabile. Levels of TCDD specifically bound in the 8-9 S peak for HepG2 and Hep3B were 27 and 34 fmol/mg cytosolic protein respectively. The level of TCDD specifically bound was not affected by charcoal treatment or by the addition of sodium molybdate, which is known to stabilize ligand binding to steroid receptors. Incubation of Hep3B or HepG2 cells with [H]TCDD at 37 degrees for 1 hr effected a redistribution of binding from the cytosol 8-9 S peak to a nuclear 6 S peak. The nuclear peaks from both human cell lines demonstrated similar sedimentation properties, temperature dependence and inducer-specificity, as for the mouse nuclear Ah receptor. Appearance of nuclear 6 S binding is consistent with a temperature-dependent translocation process, supporting the observation that these human hepatoma cell lines contain a binding component which is similar to the mouse Ah receptor in structure and function during AHH induction. PMID- 2546565 TI - Isoniazid-mediated irreversible inhibition of the myeloperoxidase antimicrobial system of the human neutrophil and the effect of thyronines. AB - During aerobic myeloperoxidase-catalysed oxidation of isoniazid at pH 7.8, compound III was generated. Oxidation of isoniazid or hydrazine sulphate at pH values of 6.5 or 7.8 in a myeloperoxidase-H2O2 system caused considerable haem loss, which was associated with compound III formation. Haem loss and also compound III formation could be inhibited when 8 microM thyroxine was included in the reaction mixtures. During the reaction with isoniazid, an intense pink coloured pigment with maximum absorbance at 500 nm was formed which could be bleached with ascorbate or hypochlorous acid. The pigment was more stable at pH 7.8 than at pH 6.5. A similar pink colour was generated when a mixture of isoniazid and thyroxine in alkaline solution was irradiated with light of wavelength greater than 300 nm. A possible product of thyroxine oxidation, 3,5 diiodotyrosine, could not protect the enzyme against isoniazid-mediated haem loss and no colour formation was observed. Haem loss was most extensive when isoniazid was oxidised in a myeloperoxidase system at pH 7.8 in the presence of 0.1 M NaCl. Thyroxine (8 microM), however, could still inhibit haem loss under these conditions. A good correlation was found between haem loss and irreversible loss of peroxidase activity. PMID- 2546566 TI - Effects of diazepam on regional brain homovanillic acid following phencyclidine or delta 9-tetrahydrocannabinol. PMID- 2546567 TI - [Molecular nature of deletion in beta 0-thalassemia, established using a method of amplification of genomic DNA in vitro]. AB - A mutation causing beta 0-thalassaemia in Azerbaijanian population is shown, by the polymerase chain reaction followed by Maxam-Gilbert sequencing, to be the deletion of dinucleotide AA from the eight codone of beta-globin gene (the mutation is known to exist also in Turkey and Lebanon). Two other mutations have also been found in beta-globin gene of the same DNA, one of which (transversion C ---G at position 16 of intron 2) eliminates the polymorphic AvaII-site and is associated with thalassaemia, and other is transition C----T in the third position of the second beta-globin codon. PMID- 2546568 TI - Detection of clonally expanded salivary gland lymphocytes in Sjogren's syndrome. AB - Recurrent swelling of the parotid and submandibular salivary glands occurs in some patients with Sjogren's syndrome (SS). Using Southern blot methods, we analyzed immunoglobulin and T cell antigen receptor (TCAR) gene rearrangements in DNA obtained from salivary gland lymphocytes of 9 SS patients. Based on histologic appearance of the biopsy specimens, these patients were diagnosed as having myoepithelial sialadenitis. We found oligoclonal rearrangements of the kappa gene (4 patients) and lambda gene (1 patient), and oligoclonal rearrangement of the TCAR beta chain in 2 additional SS patients. Patients with Ig gene rearrangements did not show rearrangements of their TCAR gene, nor did patients with TCAR rearrangements exhibit Ig rearrangements. The observed oligoclonal rearrangements probably derive from 5-10% of the salivary gland B cells or T cells. Three of these SS patients developed non-Hodgkin's lymphoma 2-8 years after the initial biopsy. Our results suggest that minor populations of B cells or T cells may clonally expand in the salivary gland tissues of SS patients with pseudolymphoma, and that such lymphocyte expansions may be controlled by the endogenous immune response and/or medications. However, continued lymphoproliferation in these salivary gland tissues may eventually lead to emergence of a neoplastic clone that escapes immunologic control and develops into a non-Hodgkin's lymphoma as a result of a multistep process. PMID- 2546569 TI - Tumor markers for early diagnosis of cholangiocarcinoma. PMID- 2546570 TI - Kinases and phosphatases in T-cell activation. AB - Protein phosphorylation-dephosphorylation plays an important role in signal transduction in T lymphocytes. In this review, Denis Alexander and Doreen Cantrell focus on the identification, regulation and functions of the kinases and phosphatases that control phosphorylation events in T cells. PMID- 2546571 TI - Structural organization of the human androgen receptor gene. AB - The complete coding region of the human androgen receptor gene has been isolated from a genomic library. The information for the androgen receptor was found to be divided over eight exons and the total length of the gene exceeded 90 kb. The sequence encoding the N-terminal region is present in one large exon. The two putative DNA-binding fingers are encoded separately by two small exons. The information for the hormone-binding domain is split over five exons. Positions of introns are identical to those reported for the chicken progesterone receptor and the human oestrogen receptor genes. Southern blot analysis of genomic DNA with various specific probes reveal that the human androgen receptor is encoded by a single-copy gene. PMID- 2546572 TI - The common occurrence of human papillomavirus infection and intraepithelial neoplasia in women infected by HIV. AB - Eighteen out of 19 women who were clinically well but infected by HIV showed abnormalities of the lower genital tract. Seven patients had intraepithelial neoplasia, histologically verified in five, involving the cervix in four, the vulva in three and the perineum in one. Lesions (clinical and/or subclinical) compatible with human papillomavirus (HPV) infection were identified in these women, as well as in 10 out of the other 11 women infected with HIV. Disease at more than one site was detected in half of the patients and would have remained covert in more than half, had colposcopy not been undertaken. Women with HIV infection are at high risk of harbouring HPV and of developing lower genital tract neoplasia. PMID- 2546573 TI - Statistics from the World Health Organization and the Centers for Disease Control. PMID- 2546574 TI - The inhibitory effects of polyoxyethylene detergents on human erythrocyte acetylcholinesterase and Ca2+ + Mg2+ ATPase. AB - The activities of acetylcholinesterase and Ca2+ + Mg2+ ATPase were measured following treatment of human erythrocyte membranes with nonsolubilizing and solubilizing concentrations of Triton X-100. A concentration of 0.1% (v/v) Triton X-100 caused a significant inhibition of both enzymes. The inhibition appears to be caused by perturbations in the membrane induced by Triton X-100 incorporation. No acetylcholinesterase activity and little Ca2+ + Mg2+ ATPase activity were detected in the supernatant at 0.05% Triton X-100 although this same detergent concentration induced changes in the turbidity of the membrane suspension. Also, no inhibition of soluble acetylcholinesterase was observed over the entire detergent concentration range. The inhibition of these enzymes at 0.1% Triton X 100 was present over an eightfold range of membrane protein in the assay indicating an independence of the protein/detergent ratio. The losses in activities of these two enzymes could be prevented by either including phosphatidylserine in the Triton X-100 suspension or using Brij 96 which has the same polyoxyethylene polar head group but an oleyl hydrophobic tail instead of the p-tert-octylphenol group of Triton X-100. The results are discussed in regard to the differential recovery of enzyme activities over the entire detergent concentration range. PMID- 2546576 TI - Silica, silicosis, and lung cancer. PMID- 2546575 TI - Vasopressin and oxytocin receptors on plasma membranes from rat mammary gland. Demonstration of vasopressin receptors by stimulation of inositol phosphate formation, and oxytocin receptors by binding of a specific 125I-labeled oxytocin antagonist, d(CH2)5(1)[Tyr(Me)2, Thr4,Tyr-NH2(9)]OVT. AB - The addition of oxytocin to minces of rat mammary gland preincubated with (3H)myo inositol stimulated the formation of inositol phosphate (IP) in both lactating and regressed glands. Stimulation was about 4 times greater in regressed tissue, consistent with an oxytocin effect on myoepithelial cells, which are enriched relative to epithelial cells during regression. The stimulation of IP formation was agonist specific, as shown with several oxytocin analogs. Arginine vasopressin (AVP), however, was more than twice as potent as oxytocin in stimulating IP formation in regressed tissue. Both V1- and V2-selective AVP receptor antagonists inhibited the stimulation of IP formation by oxytocin. The V1-selective antagonist was about 10 times more inhibitory than the V2-selective antagonist. [3H]AVP was bound to plasma membranes from the mammary gland of the lactating rat with an apparent Kd of about 0.7 nM and Bmax of 54.6 fmol/mg protein. These values were comparable with those found for AVP receptors of kidney plasma membranes. Our results suggest that the stimulation of IP formation in rat mammary gland by oxytocin occurs through occupancy of AVP, and not oxytocin, receptor sites. A second aspect of these studies was to determine if a recently developed iodinated antagonist of oxytocin-induced uterine contractions could be used as a specific probe for oxytocin receptors in the rat mammary gland. Under steady state conditions, [125I]d(CH2)5(1)[Tyr(Me)2,Thr4,Tyr NH2(9)]OVT was bound to a single class of independent binding sites in mammary gland plasma membrane from lactating rats with an apparent Kd of 65 pM and Bmax of 225 fmol/mg protein. Noniodinated antagonist had an affinity about 150 times less than the monoiodinated form. The affinity of binding sites for AVP was 10 times greater than the noniodinated antagonist and 2.4 times greater than oxytocin. In view of the presence of AVP receptors in mammary tissue, these findings suggested that the iodinated antagonist binds to AVP receptors. However, comparison of the binding of iodinated antagonist to plasma membranes from the lactating mammary gland with kidney medulla and liver, target sites for AVP, showed that binding was specific for the mammary gland and hence oxytocin receptors. The concentration of oxytocin receptors in mammary gland, as determined by [125I]d(CH2)5(1)[Tyr(Me)2,Thr4,Tyr-NH2(9)]OVT binding, was 4 times greater than the concentration of high-affinity AVP receptors, as determined by [3H]AVP binding.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546577 TI - Invasive cancer of the cervix after laser treatment. AB - Over a period of 8 years and 6 months, 1979-1988, 4222 women were treated with laser ablation for cervical intraepithelial neoplasia (CIN) with or without papillomavirus infection of the cervix. Of 3738 patients followed up, nine were diagnosed as having invasive cancers of the cervix, four of which were microinvasive. It is stressed that each woman treated with laser must have frequent, long-term cytological follow-up. PMID- 2546578 TI - Rapidly expanding exophthalmos: an unusual presentation of small cell lung cancer. AB - A 61-year-old male presented with a rapidly progressive exophthalmos from small cell lung cancer metastatic to the right orbit. His vision in that eye was 20/200, and his intraocular pressure was 36 mmHg. The orbital metastasis responded dramatically to chemotherapy. One week after starting the chemotherapy the patient did not have exophthalmos, his vision was 20/20, and three weeks later the intraocular pressure was 12 mmHg. PMID- 2546579 TI - Osmoelastic coupling in biological structures: decrease in membrane fluidity and osmophobic association of phospholipid vesicles in response to osmotic stress. AB - Poly(ethylene glycol)- (PEG-) induced change in membrane fluidity and aggregation of phospholipid vesicles were studied. A threshold concentration of PEG was required to induce the aggregation. This concentration increased with a decrease in the molecular weight of PEG, e.g., from 5% (w/w) with PEG 6000 (PEG with an average molecular weight of 7500) to more than 30% (w/w) with PEG 200. The aggregation was reversible upon dilution of PEG if the initial PEG concentration was smaller than a certain value, e.g., 22% (w/w) for PEG 6000. Addition of PEG caused a decrease in membrane fluidity of the vesicles detected by fluorescence anisotropy of diphenylhexatriene and by electron spin resonance of a spin-labeled fatty acid. The anisotropy change of diphenylhexatriene fluidity change had an inflection point at approximately 5% (w/w) of PEG 6000, which might suggest that the aggregation would make the decrease of membrane fluidity smaller. Transfer of lipid molecules between phospholipid vesicles was enhanced by the PEG-induced aggregation. The enhancement occurred not only upon direct addition of PEG to the suspending medium, but also upon dialysis of the vesicle suspension against a high concentration of PEG. All these features are consistent with osmoelastic coupling in the phospholipid membranes and the subsequent osmophobic association of the vesicles. The imbalance of osmolarity between the region adjacent to the vesicle surface (exclusion layer) and the bulk aqueous phase, which results from the preferential exclusion of PEG from the exclusion layer in the case of direct addition of PEG, exerts an osmotic stress on the vesicles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546580 TI - Elastin binds to a multifunctional 67-kilodalton peripheral membrane protein. AB - Elastin binding proteins from plasma membranes of elastin-producing cells were isolated by affinity chromatography on immobilized elastin peptides. Three proteins of 67, 61, and 55 kDa were released from the elastin resin by guanidine/detergent, soluble elastin peptides, synthetic peptide VGVAPG, or galactoside sugars, but not by synthetic RGD-containing peptide or sugars not related to galactose. All three proteins incorporated radiolabel upon extracellular iodination and contained [3H]leucine following metabolic labeling, confirming that each is a synthetic product of the cell. The 67-kDa protein could be released from the cell surface with lactose-containing buffers, whereas solubilization of the 61- and 55-kDa components required the presence of detergent. Although all three proteins were retained on elastin affinity columns, the 61- and 55-kDa components were retained only in the presence of 67-kDa protein, suggesting that the 67-kDa protein binds elastin and the 61- and 55-kDa proteins bind to the 67-kDa protein. We propose that the 67-, 61-, and 55-kDa proteins constitute an elastin-receptor complex that forms a transmembrane link between the extracellular matrix and the intracellular compartment. PMID- 2546581 TI - Biochemical identification of two types of phenamil binding sites associated with amiloride-sensitive Na+ channels. AB - The existence of distinct forms of the epithelium Na+ channel that differ in their sensitivity to amiloride has been repeatedly suggested by physiological data. The biochemical basis for these differences was analyzed by using phenamil, the most potent inhibitor known so far for the epithelium Na+ channel. [3H]Phenamil of high radioactive specific activity (30 Ci/mmol) was prepared and used to titrate [3H]phenamil binding sites in pig kidney membranes. Kinetic experiments, equilibrium binding studies, and competition experiments indicated the presence in crude membrane preparations of two classes of independent binding sites. A first binding site was characterized by a high affinity for phenamil (Kd1 = 0.4 nM) and for amiloride (Kd1 = 0.1 microM). A second binding site recognized phenamil and amiloride with lower affinities [Kd2(phenamil) = 28 nM, Kd2(amiloride) = 4 microM]. The ratio of the respective amounts of low- and high affinity binding sites was 14 +/- 2 in different membrane preparations (range: 6 22). The two types of binding sites for [3H]phenamil copurified and were still observed after purification of the epithelium Na+ channel to homogeneity. These results indicate that at least two types of pharmacologically distinguishable Na+ channels exist in the kidney. They correspond either to two isoforms of the apical Na+ channel or to one single type of channel under two different states of covalent regulation. PMID- 2546582 TI - Kinetic isotope effect studies on aspartate aminotransferase: evidence for a concerted 1,3 prototropic shift mechanism for the cytoplasmic isozyme and L aspartate and dichotomy in mechanism. AB - The C alpha primary hydrogen kinetic isotope effects (C alpha-KIEs) for the reaction of the cytoplasmic isozyme of aspartate aminotransferase (cAATase) with [alpha-2H]-L-aspartate are small and only slightly affected by deuterium oxide solvent (DV = 1.43 +/- 0.03 and DV/KAsp = 1.36 +/- 0.04 in H2O; DV = 1.44 +/- 0.01 and DV/KAsp = 1.61 +/- 0.06 in D2O). The D2O solvent KIEs (SKIEs) are somewhat larger and are essentially independent of deuterium at C alpha (D2OV = 2.21 +/- 0.07 and D2OV/KAsp = 1.70 +/- 0.03 with [alpha-1H]-L-aspartate; D2OV = 2.34 +/- 0.12 and D2OV/KAsp = 1.82 +/- 0.06 with [alpha-2H]-L- aspartate). The C alpha-KIEs on V and on V/KAsp are independent of pH from pH 5.0 to pH 10.0. These results support a rate-determining concerted 1,3 prototropic shift mechanism by the multiple KIE criteria [Hermes, J. D., Roeske, C. A., O'Leary, M. H., & Cleland, W. W. (1982) Biochemistry 21, 5106]. The large C alpha-KIEs for the reaction of mitochondrial AATase (mAATase) with L-glutamate (DV = 1.88 +/- 0.13 and DV/KGlu = 3.80 +/- 0.43 in H2O; DV = 1.57 +/- 0.05 and DV/KGlu = 4.21 +/- 0.19 in D2O) coupled with the relatively small SKIEs (D2OV = 1.58 +/- 0.04 and D2OV/KGlu = 1.25 +/- 0.05 with [alpha-1H]-L-glutamate; D2OV = 1.46 +/- 0.06 and D2OV/KGlu = 1.16 +/- 0.05 with [alpha-2H]-L-glutamate) are most consistent with a two-step mechanism for the 1,3 prototropic shift for this isozyme-substrate pair.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546583 TI - Recognition of beta beta'-substituted and alpha beta,alpha'beta'-disubstituted phosphonate analogues of bis(5'-adenosyl) tetraphosphate by the bis(5' nucleosidyl)-tetraphosphate pyrophosphohydrolases from Artemia embryos and Escherichia coli. AB - A total of 13 phosphonate analogues of bis(5'-adenosyl) tetraphosphate (AppppA) have been tested as substrates and inhibitors of the asymmetrically cleaving bis(5'-nucleosidyl) tetraphosphatase (NppppNase) from Artemia and the symmetrically cleaving NppppNase from Escherichia coli. With the Artemia enzyme, the substrate efficiency of beta beta'-substituted compounds decreased with decreasing substituent electronegativity (O greater than CF2 greater than CHF greater than CCl2 greater than CHCl greater than CH2) such that AppCF2ppA and AppCH2ppA were hydrolyzed at 70% and 2.5% of the rate of AppppA, respectively. These compounds were competitive inhibitors of this enzyme with Ki values that generally also decreased with electronegativity from 12 microM for AppCF2ppA to 0.4 microM for AppCH2ppA (Km for AppppA = 33 microM). AppCH = CHppA and AppCH2CH2ppA were neither effective substrates nor inhibitors of the Artemia enzyme. Alpha beta,alpha'beta'-Disubstituted analogues were generally less effective inhibitors with Ki values ranging from 23 microM (ApCH2ppCH2pA) to greater than 1.5 mM (ApCH2CH2ppCH2CH2pA). However, they displayed a low and unexpected rate of symmetrical cleavage by the Artemia enzyme: e.g., ApCHFppCHFpA yielded ApCHFp at 3% of the rate of AppppA breakdown. Both sets of analogues were also competitive inhibitors of the E. coli NppppNase with Ki values ranging from 7 microM (AppCH2ppA) to 250 microM (ApCH2CH2ppCH2CH2pA) (Km for AppppA = 28 microM). The only alpha beta,alpha'beta'-disubstituted analogue to be hydrolyzed by the E. coli enzyme was ApCF2ppCF2pA at 0.2% of the rate of AppppA; however, several of the beta beta'-substituted compounds showed a limited degree of asymmetrical cleavage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546584 TI - Mechanism of inhibition of DNA gyrase by quinolone antibacterials: specificity and cooperativity of drug binding to DNA. AB - Although the functional target of quinolone antibacterials such as nalidixic acid and norfloxacin has been identified as the enzyme DNA gyrase, the direct binding site of the drug is the DNA molecule [Shen, L. L., & Pernet, A. G. (1985) Proc. Natl. Acad. Sci. U.S.A. 82, 307-311]. As described in this paper, binding specificity and cooperativity of quinolones to DNA were further investigated with the use of a variety of DNA species of different structures and different base compositions. Results show that the drug binding specificity is controlled and determined largely by the DNA structure. The drug binds weakly and demonstrates no base preference when DNA strands are paired. The drug binds with much greater affinity when the strands are separated, and consequently, binding preference emerges: it binds better to poly(G) and poly(dG) over their counterparts including poly(dI). The results suggest that the drug binds to unpaired bases via hydrogen bonding and not via ring stacking with DNA bases. The weak binding to relaxed double-stranded DNA and the stronger binding to single-stranded DNA are both nonspecific as they do not demonstrate binding saturation and cooperativity. The specific type of binding, initially demonstrated in our previous publication with the supercoiled DNA and more recently with complex formed between linear DNA and DNA gyrase [Shen, L. L., Kohlbrenner, W. E., Weigl, D., & Baranowski, J. (1989) J. Biol. Chem. (in press)], occurs near the drug's supercoiling inhibition concentration. As shown in this paper, binding saturation curves of this type are highly cooperative (with Hill constant greater than 4).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546585 TI - Mechanism of inhibition of DNA gyrase by quinolone antibacterials: a cooperative drug--DNA binding model. AB - We have proposed a cooperative quinolone-DNA binding model for the inhibition of DNA gyrase. The essential feature of the model is that bound gyrase induces a specific quinolone binding site in the relaxed DNA substrate in the presence of ATP. The binding affinity and specificity are derived from two unique and equally important functional features: the specific conformation of the proposed single stranded DNA pocket induced by the enzyme and the unique self-association phenomenon (from which the cooperativity is derived) of the drug molecules to fit the binding pocket with a high degree of flexibility. Supporting evidence for and implications of this model are provided. PMID- 2546586 TI - Modulation of mitomycin cross-linking by DNA bending in the Escherichia coli CAP protein-DNA complex. AB - We have examined the comparative reactivity of mitomycin cross-linking sites in DNA molecules either free in solution or complexed with Escherichia coli CAP protein. Sites in the region to which the protein is bound show strongly variable cross-linking by the drug. The reactivity of a CpG site located where the minor groove is narrowed by bending toward the protein was decreased by about 4-fold, compared to free DNA. The reactivity of a site placed so that the minor groove is widened by the bend was reduced by about 25%, and the reactivity of a (CpG)3 sequence facing primarily away from the protein was reduced 25-fold by CAP binding. These results support the view that local DNA structure plays a critical role in determining the efficiency of cross-linking. PMID- 2546587 TI - Long-lived tryptophan fluorescence in phosphoglycerate mutase. AB - Phosphoglycerate mutase (PGM; EC 2.7.5.3) isolated from rat and rabbit muscle has been shown to possess an unusually long-lived fluorescence component when excited by ultraviolet light below 310 nm. On the basis of spectral and physical measurements, this 16.4 (+/- 0.2) ns fluorescence lifetime at room temperature is assigned to a tryptophan residue in an unusual environment. The emission profile of this long-lived tryptophan is red shifted from the other tryptophans of PGM by approximately 25 nm. PGM has been crystallized and sequenced from yeast where it has been shown to be a tetramer with 29K subunits. However, we have not been able to detect the existence of an unusually long-lived fluorescence component in the yeast isomer. The long fluorescence lifetime is lost upon denaturation of rabbit PGM and is partially restored upon introduction of the protein to a nondenaturing environment, suggesting the long lifetime is not the result of a covalent modification. The PGM molecule was studied by a number of techniques including time-resolved tryptophan fluorescence, quenching studies of tryptophan fluorescence, and enzyme activity studies. The long-lived fluorescence has been shown to be statistically quenched by Br-, I-, and Cu2+ in the submillimolar region while the acrylamide quenching shows the tryptophan is marginally accessible to solvent. Characterization of the long-lived fluorescence and its possible sources are discussed. PMID- 2546588 TI - Magnetic circular dichroism and electron paramagnetic resonance studies of iron(II)-metallothionein. AB - The electronic and magnetic properties of the Fe(II)-thiolate centers in Fe(II) metallothionein have been investigated by low-temperature magnetic circular dichroism and electron paramagnetic resonance spectroscopies at various levels of Fe(II) incorporation. In agreement with previous results [Good, M., & Vasak, M. (1986) Biochemistry 25, 8353-8356], rabbit liver metallothionein was found to bind a maximum of seven Fe(II) ions, with cluster formation occurring when more than four Fe(II) ions are bound at pH 8.5. The results indicate that all the iron in fully loaded Fe(II)-metallothionein is accommodated in Fe(II)-thiolate clusters that have either S = 0 or S = 2 ground states as a result of antiferromagnetic coupling between high-spin Fe(II) ions. By analogy with the cluster composition and mechanism of assembly that have been established for other divalent metal ions, the clusters with S = 0 and S = 2 ground states are attributed to tetranuclear and trinuclear centers, respectively. EPR signals indicative of S = 2 species were observed for samples containing monomeric tetrathiolate-Fe(II) centers and trinuclear Fe(II)-thiolate clusters. However, the nature of the zero-field splitting of the S = 2 ground states that is indicated by the EPR signals is not consistent with that deduced from Mossbauer and magnetic circular dichroism studies, suggesting heterogeneity in both types of center. PMID- 2546589 TI - Localization of the fourth membrane spanning domain as a ligand binding site in the human platelet alpha 2-adrenergic receptor. AB - The human platelet alpha 2-adrenergic receptor is an integral membrane protein which binds epinephrine. The gene for this receptor has been cloned, and the primary structure is thus known [Kobilka et al. (1987) Science 238, 650-656]. A model of its secondary structure predicts that the receptor has seven transmembrane spanning domains. By covalent labeling and peptide mapping, we have identified a region of the receptor that is directly involved with ligand binding. Partially purified preparations of the receptor were covalently radiolabeled with either of two specific photoaffinity ligands: [3H]SKF 102229 (an antagonist) or p-azido[3H]clonidine (an agonist). The radiolabeled receptors were then digested with specific endopeptidases, and peptides containing the covalently bound radioligands were identified. Lysylendopeptidase treatment of [3H]SKF 102229 labeled receptor yielded one peptide of Mr 2400 as the product of a complete digest. Endopeptidase Arg-C gave a labeled peptide of Mr 4000, which was further digested to the Mr 2400 peptide by additional treatment with lysylendopeptidase. Using p-azido[3H]clonidine-labeled receptor, a similar Mr 2400 peptide was obtained by lysylendopeptidase cleavage. This Mr 2400 peptide corresponds to the fourth transmembrane spanning domain of the receptor. These data suggest that this region forms part of the ligand binding domain of the human platelet alpha 2-adrenergic receptor. PMID- 2546590 TI - On the relationship between leukotriene and lipoxin production by human neutrophils: evidence for differential metabolism of 15-HETE and 5-HETE. AB - Lipoxygenase (LO) products generated by human PMN were examined utilizing a gradient-HPLC and rapid spectral detector which permitted continuous UV-spectral monitoring of leukotrienes, lipoxins and related oxygenated products of arachidonic acid. When exposed to the ionophore A23187, PMN generated LTB4 and its omega-oxidation products as well as LXA4, LXB4, and 7-cis-11-trans-LXA4 from endogenous sources. Addition of 15-HETE changed the profile of products generated by activated PMN and led to a time- and dose-dependent increase in lipoxins and related compounds while the production of LTB4 and its omega-oxidation products was inhibited. Results of time-course and radiolabel studies revealed that 15 HETE is rapidly transformed within 15 s to 5,15-DHETE and conjugated tetraene containing products, and that the inhibition of leukotriene formation followed a similar time-course. In contrast, PMN did not generate either lipoxins or related products from 5-[3H]HETE, nor did 5-HETE block leukotriene formation. Stimulated PMN generated 5,15-DHETE from exogenous 5-HETE, while in the absence of ionophore, 5-HETE was transformed to 5,20-HETE. These results indicate that PMN can generate lipoxins and related products from endogenous sources and that 15 HETE and 5-HETE are transformed by different routes. PMID- 2546591 TI - Differences in the metabolism of inositol and phosphoinositides by cultured cells of neuronal and glial origin. AB - Phosphoinositide and inositol metabolism was compared in glioma (C6), neuroblastoma (N1E-115) and neuroblastoma X glioma hybrid (NG 108-15) cells. All cell lines had similar proportions of phosphatidylinositol (PI), phosphatidylinositol 4-phosphate (PIP), and phosphatidylinositol 4,5-bisphosphate (PIP2). Neuroblastoma and hybrid cells had almost identical phospholipid and phosphoinositide compositions and similar activities for the enzymes metabolizing polyphosphoinositides (PI kinase, PIP phosphatase, PIP kinase, PIP2 phosphatase, PIP2 phosphodiesterase). Glioma cells differed by having greater proportions of ethanolamine plasmalogen and sphingomyelin, lower PIP kinase, 3-5-fold higher PIP phosphatase activity and 10-15-fold greater PIP2 phosphodiesterase activity. Higher PIP phosphatase and PIP2 diesterase activities appear to be characteristic of cells of glial origin, since similar activities were found in primary cultures of astroglia. Glioma cells also metabolize inositol differently. In pulse and pulse-chase experiments, glioma cells transported inositol into a much larger water-soluble intracellular pool and maintained a concentration gradient 30-times greater than neuroblastoma cells. Label in intracellular inositol was less than in phosphoinositides in neuroblastoma and exchanged rapidly with extracellular inositol. In glioma, labeling of intracellular inositol greatly exceeded that of phosphoinositides. As a consequence, radioactivity in prelabeled phosphoinositides could not be effectively chased from glioma cells by excess unlabeled inositol. Such differences between cells of neuronal and glial origin suggest different and possibly supportive roles for these two cell types in maintaining functions regulated through phosphoinositide-linked signalling systems in the central nervous system. PMID- 2546592 TI - In vitro phosphorylation of phosphatidylethanolamine N-methyltransferase by cAMP dependent protein kinase: lack of in vivo phosphorylation in response to N6-2'-O dibutryladenosine 3',5'-cyclic monophosphate. AB - Phosphorylation of rat liver phosphatidylethanolamine (PE) N-methyltransferase by cAMP-dependent protein kinase was investigated. The 18 kDa methyltransferase was found to be phosphorylated in vitro by cAMP-dependent protein kinase on a serine residue. The stoichiometry of phosphate incorporation reached a maximum of 0.25 mol phosphate/mol methyltransferase at 30 min. Resolution of the phosphorylated methyltransferase by two-dimensional gel electrophoresis showed that two isoproteins were substrates. Phosphorylation of the purified PE N methyltransferase for up to 1 h had no effect on the methylation of PE, PMME or PDME. To test for in vivo phosphorylation, isolated rate hepatocytes were exposed to 0.5 mM N6-2'-O-dibutryladenosine 3':5'-cyclic monophosphate (DiB-cAMP) and the phosphorylation state of microsomal proteins evaluated by two-dimensional gel electrophoresis, nitrocellulose blotting and autoradiography. The same nitrocellulose blots were probed with a rabbit anti-PE N-methyltransferase antibody, immunochemically stained and aligned with the autoradiogram. No phosphorylated proteins co-migrated with the methyltransferase under non phosphorylating conditions, or when hepatocytes were exposed to the cAMP analogue for up to 2 h. Oddly, DiB-cAMP increased both PE- and PMME-dependent activity in isolated microsomes, but decreased PE to PC conversion measured in intact hepatocytes. The results indicated that PE N-methyltransferase is poorly phosphorylated by cAMP-dependent protein kinase in vitro, and is not phosphorylated in intact hepatocytes treated with a cAMP analogue. PMID- 2546593 TI - The effect of polycyclic aromatic hydrocarbons on choline kinase activity in mouse hepatoma cells. AB - Choline kinase catalyzes the first rate-limiting step in the pathway of biosynthesis of phosphatidylcholine. This enzyme was shown previously to be induced in liver by treatment of rats with polycyclic aromatic hydrocarbons (Ishidate et al. (1980) Biochem. Biophys. Res. Commun. 96, 946-952). The present study was undertaken to determine whether choline kinase in the murine hepatoma cell line, Hepa 1c1c7, is inducible by aromatic hydrocarbons and, if so, whether this induction is mediated by the aromatic hydrocarbon receptor. Treatment of Hepa 1c1c7 cells with 10 microM beta-naphthoflavone resulted in a 1.6-fold increase of choline kinase activity, but no response was seen when the cells were exposed to either 5.0 microM benzo[a]pyrene or 1.0 nM 2.3,7,8-tetrachlorodibenzo p-doxin, both potent inducers of aryl hydrocarbon hydroxylase. Cell line variants with either deficient or elevated aromatic hydrocarbon receptors showed no increase in choline kinase activity following treatment with any of the polycyclic aromatic hydrocarbons. These results are not consistent with a role for the aromatic hydrocarbon receptor in increased choline kinase activity in Hepa 1c1c7 cells. PMID- 2546594 TI - Reorientational dynamics in lipid vesicles and liposomes studied with ESR: effects of hydration, curvature and unsaturation. AB - Electron spin resonance experiments were carried out on 3-doxyl-5 alpha cholestane spin-label (CSL) molecules embedded in multilamellar liposomes and small unilamellar vesicles (SUVs) of palmitoyloleoylphosphatidylcholine (POPC), dioleoylphosphatidylcholine (DOPC) and dilinoleoylphosphatidylcholine (DLPC). The experimental spectra were analyzed by a numerical solution of the stochastic Liouville equation. Effects of temperature, presence of unsaturated bonds and high bilayer curvature on the dynamic behaviour of the lipid molecules were studied. Our results, combined with results from planar multibilayers with a varying hydration rate (Korstanje et al. (1989) Biochim. Biophys. Acta 980, 225 233), give a consistent picture of the orientational order and rotational dynamics of CSL molecules embedded in lipid matrices with various geometrical configurations. Increase of hydration or temperature reduces molecular ordering and increases molecular dynamics. In highly curved vesicle configurations, SUVs, molecular order is found to be lower than in multilamellar liposomes. In contrast, rotational motion is not affected by increase of curvature. In all lipid configurations studied, increase of the number of unsaturated bonds in the fatty acid chains reduces molecular ordering. We find, however, no effect of unsaturation on the rotational mobility of the CSL probe molecules. These results clearly show that changes in molecular orientational order and reorientational dynamics have to be considered separately, and that they are not necessarily correlated as implied by the common concept of membrane fluidity. Comparing our results with data from a motional narrowing analysis shows that the latter approach seriously overestimates the rate of molecular reorientation. PMID- 2546595 TI - Comparison of the effects of NaCl on the thermotropic behaviour of sn-1' and sn 3' stereoisomers of 1,2-dimyristoyl-sn-glycero-3-phosphatidylglycerol. AB - The phase behaviour of liposomes of 1,2-dimyristoyl-sn-glycero-3-phosphatidyl-sn 1'-glycerol (1'-DMPG) and the corresponding sn-3' stereoisomer (3'-DMPG) were studied by DSC as a function of NaCl concentration. The melting of the metastable gel phase to the liquid-crystalline phase was similar for both lipids. However, in the presence of salt and at 6 degrees C (T less than Tp) the gel phase of both stereoisomers of DMPG was shown to be metastable and a new phase nominated here as the highly crystalline phase was formed as the stable state. However, significant differences in the formation and melting of the highly crystalline phase were evident between the two polar headgroup stereoisomers. For 3'-DMPG in the presence of 300 mM NaCl the melting enthalpy of this phase is approx. 82 kJ/mol and the transition temperature about 11 degrees higher (at 33.6 degrees C) than for the gel to liquid-crystalline phase transition (25 kJ/mol at 23.0 degrees C). In the presence of 0.15-1.2 M NaCl at 6 to 10 degrees C the formation of the highly crystalline phase of 3'-DMPG is complete within 2 to 5 days, increasing [NaCl] facilitates the rate. For a 1:1 mixture of 1'- and 3'-DMPG the formation of the highly crystalline phase requires several weeks and melts at about 20 degrees higher than the gel phase (at approx. 40 degrees C). For 1'-DMPG partial conversion into the highly crystalline phase requires several months. For 3'-DMPG several intermediate phases appeared as endothermic peaks between the main phase transition temperature and the melting temperature of the highly crystalline phase. In contrast, for 1'-DMPG and the 1:1 mixture the subgel phase appears to be the only metastable intermediate phase. Different monovalent cations differ in their effect on the metastable behaviour. PMID- 2546596 TI - Galactoside-dependent proton transport by mutants of the Escherichia coli lactose carrier: substitution of tyrosine for histidine-322 and of leucine for serine 306. AB - The lac Y genes from two Escherichia coli mutants, MAB20 and AA22, have been cloned in a multicopy plasmid by a novel 'sucrose marker exchange' method. Characterization showed that the plasmids express a lactose carrier with poor affinity for lactose. Neither mutant carried out concentrative uptake with methyl beta-D-galactopyranoside, lactose, or melibiose as the substrate. Nor did the mutants catalyze counterflow or exchange with methyl beta-D-galactopyranoside. Both mutants did, however, retain the capacity to carry out facilitated diffusion with lactose or melibiose. DNA sequencing revealed that MAB20 (histidine-322 to tyrosine) and AA22 (serine-306 to leucine) have amino acid substitutions within the putative 'charge-relay' domain thought to be responsible for proton transport. Galactoside-dependent H+ transport was readily measured in both mutants. We conclude, therefore, that the presence of a histidine residue at position 322 of the lactose carrier is not obligatory for H+ transport per se. PMID- 2546597 TI - Effects of Ca2+ on the sodium pump observed in cardiac myocytes isolated from guinea pigs. AB - It is presently unknown whether Ca2+ plays a role in the physiological control of Na+/K+-ATPase or sodium pump activity. Because the enzyme is exposed to markedly different intra- and extracellular Ca2+ concentrations, tissue homogenates or purified enzyme preparations may not provide pertinent information regarding this question. Therefore, the effects of Ca2+ on the sodium pump were examined with studies of [3H]ouabain binding and 86Rb+ uptake using viable myocytes isolated from guinea-pig heart and apparently maintaining ion gradients. In the presence of K+, a reduction of the extracellular Ca2+ increased specific [3H]ouabain binding observed at apparent binding equilibria: a half-maximal stimulation was observed when extracellular Ca2+ was lowered to about 50 microM. The change in [3H]ouabain binding was caused by a change in the number of binding sites accessible by ouabain instead of a change in their affinity for the glycoside. Ouabain-sensitive 86Rb+ uptake was increased by a reduction of extracellular Ca2+ concentration. Benzocaine in concentrations reported to reduce the rate of Na+ influx failed to influence the inhibitory effect of Ca2+ on glycoside binding. When [3H]ouabain binding was at equilibrium, the addition of Ca2+ decreased and that of EGTA increased the glycoside binding. Mn2+, which does not penetrate the cell membrane, had effects similar to Ca2+. In the absence of K+, cells lose their tolerance to Ca2+. Reducing Ca2+ concentration prevented the loss of rod shaped cells but failed to affect specific [3H]ouabain binding observed in the absence of K+. These results indicate that a large change in extracellular Ca2+ directly affects the sodium pump in cardiac myocytes isolated from guinea pigs. PMID- 2546598 TI - Enhanced number of actin binding sites on plasma membranes of polyoma virus transformed fibroblasts. AB - Plasma membranes, isolated from normal (C13) and polyoma virus-transformed (J1) cultured BHK cells were incubated with G-actin under polymerizing conditions, followed by a low-speed centrifugation. The amount of actin attached to the pelleted BHK-J1 plasma membranes was at least twice that on BHK-C13 membranes, indicating a greater number of actin attachment sites on the former. This result was confirmed by the observation that the plasma membranes from the transformed cells were also more active in nucleating polymerization of pyrene-labelled actin. Most of the actin attachment sites could be solubilized by Triton or low salt extraction treatment. PMID- 2546599 TI - Structure and function of platelet-derived growth factor (PDGF) and related proteins. PMID- 2546600 TI - Control of the higher eukaryote cell cycle by p34cdc2 homologues. PMID- 2546601 TI - The products of the reduction of doxyl stearates in cells are hydroxylamines as shown by oxidation by 15N-perdeuterated Tempone. AB - The use of nitroxides in functional biological systems has increased greatly as it has become evident that such studies can provide valuable biophysical and metabolic data. This has led to a need to understand the nature of the metabolism of nitroxides and their products. This paper presents data indicating the value of 15N-perdeuterated Tempone specifically to indicate the amount of hydroxylamines that are present in a cellular system. Using this technique, we found that in the mammalian cells that we studied the principal or only products of reduction of doxyl stearates were the corresponding hydroxylamines. PMID- 2546602 TI - Activation of pyridoxal kinase by metallothionein. AB - Brain pyridoxal kinase, which uses ATP complexed to either Zn(II) or Co(II) as substrates, displays high catalytic activity in the presence of Zn-thionein and Co-thionein. Several steps intervene in the process of pyridoxal kinase activation, i.e., binding of Zn ions to ATP and interaction between Zn-ATP and the enzyme. Equilibrium binding studies show that ATP mediates the release of Zn ions from the metal-thiolate clusters of the thioneins, whereas spectroscopic measurements conducted on Co-thionein reveal that the absorption transitions corresponding to the metal-thiolate of the protein are perturbed by ATP. The binding Zn-ATP to the kinase proceeds with a delta G = -6.3 kcal/mol as demonstrated by fluorometric titrations. Direct interaction between the kinase and derivatized-metallothionein could not be detected by emission anisotropy measurements, indicating that juxtaposition of the proteins does not influence the exchange of metal ions. Since the concentration of free Zn in several mammalian tissues is lower than 1 nM, it is postulated that under in vivo conditions the concentration of metallothionein regulates the catalytic activity of pyridoxal kinase. PMID- 2546603 TI - Proton NMR investigation of the influence of subunit assembly on the low-spin in equilibrium high-spin equilibrium of met-azido hemoglobin A. AB - The 1H nuclear magnetic resonance signals for the side-chain labile protons of the proximal His-F8 in met-azido derivatives of the isolated chains and intact tetramer of hemoglobin have been identified. Assignment of the two peaks to the individual subunits of the intact tetramer was effected by the basis of the strong similarity of shift of one of the two peaks to that of met-azido semi hemoglobin, where hemes occupy primarily the alpha subunits, with the heme cavity vacant in the adjacent beta subunits. The magnitudes of the hyperfine shift for both the His-F8 ring NH and the heme methyls reflect the degree of high-spin character in the thermal spin equilibrium between the high-spin, S = 5/2, and low spin, S = 1/2, states. The changes in these shifts upon tetramer assembly demonstrate that formation of the intersubunit contacts in the R-state met-azido hemoglobin from the isolated chains causes a slight decrease in high-spin character of the alpha (22 to 20%) and a marked increase (5 to 11%) in the high spin character of the beta subunits. The changes in spin-character are interpreted in terms of slight increase and decrease in the strength of the iron His F8 bond upon tetramer assembly in the alpha and beta subunits, respectively. These changes in axial bonding upon forming R-state intersubunit contacts are consistent with previous observation on forming the R-state deoxy Hb tetramer from the isolated chains (Nagai, K., La Mar, G.N., Jue, T. and Bunn, H.F. (1982) Biochemistry 21, 842-847). PMID- 2546604 TI - Binding of aromatic donor molecules to lactoperoxidase: proton NMR and optical difference spectroscopic studies. AB - The interaction of aromatic donor molecules with lactoperoxidase (LPO) was studied using 1H-NMR and optical difference spectroscopy techniques. pH dependence of substrate proton resonance line-widths indicated that the binding was facilitated by protonation of an amino acid residue (with pKa of 6.1) which is presumably a distal histidine. Dissociation constants evaluated from both optical difference spectroscopy and 1H-NMR relaxation measurements were found to be an order of magnitude larger than those for binding to horse radish peroxidase (HRP), indicating relatively weak binding of the donors to LPO. The dissociation constants evaluated in presence of excess of I- and SCN- showed a considerable increase in their values, indicating that the iodide and thiocyanate ions compete for binding at the same site. The dissociation constant of the substrate binding was, however, not affected by cyanide binding to the ferric centre of LPO. All these results indicate that the organic substrates bind to LPO away from the ferric center. Comparison of the dissociation constants between the different substrates suggested that hydrogen bonding of the donors with the distal histidine amino acid, and hydrophobic interaction between the donors and the active site contribute significantly towards the associating forces. Free energy, entropy and enthalpy changes associated with the LPO-substrate equilibrium have been evaluated. These thermodynamic parameters were found to be all negative and relatively low compared to those for binding to HRP. The distances of the substrate protons from the ferric center were found to be in the range 9.4-11.1 A which are 2-3 A larger than those reported for the HRP-substrate complexes. These structural informations suggest that the heme in LPO may be more deeply buried in the heme crevice than that in the HRP. PMID- 2546606 TI - Formation of cyanoferricytochrome c in the presence of potassium ferricyanide, in the course of a Raman resonance experiment. AB - Modifications of native ferricytochrome c are observed in the course of Raman resonance experiments, run in the presence of residual amounts of potassium ferricyanide. Visible and EPR data shown that the cyanide-substituted cytochrome is formed, due to photodecomposition of ferricyanide. Its Raman resonance spectrum is reported. PMID- 2546605 TI - Stimulation by glycerate 2,3-bisphosphate: a common property of cytosolic IMP-GMP 5'-nucleotidase in rat and human tissues. AB - Glycerate 2,3-bisphosphate, a potent stimulator of the cytosolic 5'-nucleotidase which preferentially hydrolyzes IMP and GMP in human erythrocytes (Bontemps et al., 1988, Biochem. J. 250, 687-696), also stimulates the dephosphorylation of IMP in cytosol fractions of rat heart, liver, brain, kidney, spleen and erythrocytes, and of human polymorphonuclear leucocytes, mixed peripheral blood lymphocytes, platelets and fibroblasts. Depending on the cell type, stimulation by 5 mM glycerate 2,3-bisphosphate varied from 1.5- to 12-fold. Where investigated, glycerate 2,3-bisphosphate had an approx. 5-fold higher affinity for the enzyme than its other stimulator, ATP. These observations provide a useful tool to distinguish IMP-GMP 5'-nucleotidase from other 5'-nucleotidases, and suggest a common origin of the cytosolic IMP-GMP 5'-nucleotidase in various tissues. PMID- 2546607 TI - Conformation of bovine nitrosylhemoglobins: an ESR study. AB - The structural properties of nitrosylhemoglobins from two bovine species, namely cow and buffalo, have been investigated using electron spin resonance spectroscopy. Bovine hemoglobins show sensitivity to the presence of chloride ions and organic phosphates. ESR spectral features indicate a stable deoxy quaternary conformation of the molecule when compared to normal adult human hemoglobin A. Amino acid substitutions at the amino terminal end of the beta chain and at other sites of the alpha and beta chains seems to shift the allosteric equilibrium towards the T state in bovine hemoglobins. The results also confirm the intrinsically low oxygen affinity of bovine hemoglobins under physiological conditions. PMID- 2546608 TI - Modification of ligand binding to the Na+/K+-activated ATPase. AB - Interactions between the ligands Mg2+, K+, and substrate and the Na+/K+-activated ATPase were examined in terms of a rapid-equilibrium, random-order, terreactant kinetic scheme for the K+-nitrophenyl phosphatase reaction that is catalyzed by this enzyme. At 37 degrees C and pH 7.5 the derived values for the dissociation constants from the free enzyme were 0.2, 0.08, and 1.4 mM for Mg2+, K+, and substrate, respectively. For Mg2+ interactions, the presence of 20% (v/v) dimethyl sulfoxide (Me2SO) increased the calculated affinity 25-fold; higher concentrations increased affinity still further. Neither reducing the temperature to 20 degrees C nor altering the pH from 6.5 to 8.3 appreciably changed the affinity for Mg2+ in the absence or presence of Me2SO. The Mg2+ sites are thus characterized by an absence of functional groups ionizable in the pH range 6.5 8.3, with binding driven by entropy changes, and with Me2SO, probably through solvation effects on the protein, increasing affinity for Mg2+ close to that for Ca2+ and Mn2+. By contrast, for K+ interactions, the presence of 20% Me2SO increased the calculated affinity only by half; moreover, reducing the temperature to 20 degrees C and the pH to 6.5 both increased affinity and diminished the response to Me2SO. The K+ sites are thus characterized by a marked sensitivity to pH and temperature, presumably through alterations in enzyme conformational equilibria that in turn are modifiable by Me2SO. Inhibition by higher concentrations of Mg2+, which varies inversely with the K+ concentration, was decreased by Me2SO. Finally, for substrate interactions, the presence of 20% Me2SO increased the calculated affinity 4-fold, and, as for Mg2+-binding, neither reducing the temperature nor varying the pH over the range 6.5-8.3 appreciably altered the affinity in the absence or presence of Me2SO. Thus, the substrate sites, like the Mg2+ sites, are characterized by an absence of functional groups ionizable in this range, with binding driven by entropy changes, and with Me2SO increasing affinity for substrate, in this case probably through favoring the partitioning of substrate from the medium into the hydrophobic active site. PMID- 2546610 TI - Catalytic mechanism and interactions of NAD+ with glyceraldehyde-3-phosphate dehydrogenase: correlation of EPR data and enzymatic studies. AB - Perdeuterated spin label (DSL) analogs of NAD+, with the spin label attached at either the C8 or N6 position of the adenine ring, have been employed in an EPR investigation of models for negative cooperativity binding to tetrameric glyceraldehyde-3-phosphate dehydrogenase and conformational changes of the DSL NAD+-enzyme complex during the catalytic reaction. C8-DSL-NAD+ and N6-DSL-NAD+ showed 80 and 45% of the activity of the native NAD+, respectively. Therefore, these spin-labeled compounds are very efficacious for investigations of the motional dynamics and catalytic mechanism of this dehydrogenase. Perdeuterated spin labels enhanced spectral sensitivity and resolution thereby enabling the simultaneous detection of spin-labeled NAD+ in three conditions: (1) DSL-NAD+ freely tumbling in the presence of, but not bound to, glyceraldehyde-3-phosphate dehydrogenase, (2) DSL-NAD+ tightly bound to enzyme subunits remote (58 A) from other NAD+ binding sites, and (3) DSL-NAD+ bound to adjacent monomers and exhibiting electron dipolar interactions (8-9 A or 12-13 A, depending on the analog). Determinations of relative amounts of DSL-NAD+ in these three environments and measurements of the binding constants, K1-K4, permitted characterization of the mathematical model describing the negative cooperativity in the binding of four NAD+ to glyceraldehyde-3-phosphate dehydrogenase. For enzyme crystallized from rabbit muscle, EPR results were found to be consistent with the ligand-induced sequential model and inconsistent with the pre-existing asymmetry models. The electron dipolar interaction observed between spin labels bound to two adjacent glyceraldehyde-3-phosphate dehydrogenase monomers (8-9 or 12-13 A) related by the R-axis provided a sensitive probe of conformational changes of the enzyme-DSL-NAD+ complex. When glyceraldehyde-3-phosphate was covalently bound to the active site cysteine-149, an increase in electron dipolar interaction was observed. This increase was consistent with a closer approximation of spin labels produced by steric interactions between the phosphoglyceryl residue and DSL-NAD+. Coenzyme reduction (DSL-NADH) or inactivation of the dehydrogenase by carboxymethylation of the active site cysteine-149 did not produce changes in the dipolar interactions or spatial separation of the spin labels attached to the adenine moiety of the NAD+. However, coenzyme reduction or carboxymethylation did alter the stoichiometry of binding and caused the release of approximately one loosely bound DSL-NAD+ from the enzyme. These findings suggest that ionic charge interactions are important in coenzyme binding at the active site. PMID- 2546609 TI - Catecholamine melanins. Structural changes induced by copper ions. AB - Melanins synthesized from adrenaline and dopamine in the presence or absence of copper ions were characterized by pyrolysis-gas chromatography-mass spectrometry and by IR and ESR methods. It was shown that Cu2+ are able to induce changes in the melanin structure. Melanins obtained from adrenaline-Cu2+ and dopamine-Cu2+ complexes are composed mainly from monomeric units of the indole type. Melanins synthesized from these catecholamines without Cu2+ contain additionally large amounts of unindolized monomeric units. The structure differences in both types of melanins are reflected in their sorptive abilities and spectroscopic characteristics. PMID- 2546611 TI - [Solubilization of the central benzodiazepine receptor and study of its interaction with nicotinamide]. AB - It was shown that nicotinamide and NAD inhibit the specific binding of [3H]flunitrazepam to benzdiazepine receptors without causing a direct influence of gamma-aminobutyric acid (GABA) receptors. The GABA-benzdiazepine complex was separated by solubilization with 0.5% lubrol PX. The solubilized preparations contain the binding sites for [3H]flunitrazepam alone (Kd = 5.9 nm). The Kd value for the membrane-bound benzdiazepine receptor is 2.9 nM. NAD inhibited the specific binding of [3H]flunitrazepam to the solubilized membrane preparation when used at concentrations by several orders of magnitude lower than that of nicotinamide. Using gel filtration on Sepharose 6B-CL, the molecular mass of the soluble benzdiazepine receptor protein was determined. PMID- 2546613 TI - [Lack of negative regulation of beta-adrenoreceptor affinity by guanine nucleotides in embryonic membranes and its induction by the GTP-binding protein from mature protein]. AB - The negative regulation of the beta-adrenoreceptor affinity by guanine nucleotides in the sarcolemmal fraction of chicken skeletal muscle at different stages of ontogenesis was studied. It was found that the negative regulation is absent in the embryonic period; the effect of GTP is manifested only before hatching, whereas that of Gpp(NH)p--at later periods, i.e., in 1-month-old chickens. Similar age-dependent dynamics was revealed with respect to the GTP effect on the dissociation rate of the [3H]DHA-beta-adrenoreceptor complex. An addition to the system containing embryonic muscle membranes of the GTP-binding protein isolated from skeletal muscle and liver of chickens whose age exceeds 20 days led to earlier manifestations of the above effects (on the 13th-15th embryonic days). The data obtained testify to the limiting role of GTP-binding proteins in the negative control of the hormone-receptor interaction and support the authors' hypothesis on the absence in the embryonic muscle of the 42 kD GTP binding protein responsible for the functional coupling of the hormone-sensitive adenylate cyclase components. PMID- 2546612 TI - [Substrate specificity of multiple forms of human alpha-D-galactosidase and alpha D-fucosidase]. AB - It was shown that human alpha-D-galactosidase is represented by multiple forms, only one of which can also split alpha-D-fucoside. Fabry's disease was found to be associated not only with the deficiency of the alpha-D-galactosidase total activity but also with the deficiency of the alpha-D-fucosidase activity. The decrease in the alpha-D-galactosidase activity is due to the lack of two enzyme forms, while the profile of alpha-D-fucosidase multiple forms during isoelectric focusing of human enzyme preparations is modified very little in comparison with the normal one. The deficiency of both enzymes was expressed in most degree in leukocytes as compared to other tissues. The residual activities of alpha-D galactosidase and alpha-D-fucosidase in leukocytes were equal to 3.5 and 21%, respectively. Since the decrease in the alpha-D-fucosidase activity was not so noticeable as in the alpha-D-galactosidase activity, it may be expected that the determination of the alpha-D-fucosidase activity can no longer be regarded as a reliable test for the diagnosis of Fabry's disease. The data obtained suggest that alpha-D-galactoside and alpha-D-fucoside are split by the same enzyme, the multiple forms of which are characterized by selective specificity towards these substrates. PMID- 2546614 TI - The role of inositol lipid metabolism in the ovary. AB - Two major signal transduction systems operate within ovarian cells to control their function. Gonadotropins, such as follicle-stimulating hormone and luteinizing hormone, primarily utilize the cyclic adenosine 3',5'-monophosphate (cAMP) pathway to stimulate steroid hormone biosynthesis. On the other hand, an inositol lipid metabolism pathway is used by other effector molecules such as gonadotropin-releasing hormone or prostaglandin F2 alpha, as well as gonadotropins, to alter ovarian hormone production. Membrane polyphosphoinositides are hydrolyzed to inositol phosphates and diacylglycerol, resulting in alterations of intracellular free calcium concentration, activation of protein kinase C, and liberation of arachidonic acid. Some or all of these intracellular messengers may interact with the gonadotropin-induced cAMP pathway to control ovarian function. PMID- 2546615 TI - Differential effects of the destruction of Leydig cells by administration of ethane dimethane sulphonate to postnatal rats. AB - Ethane dimethane sulphonate (EDS) is a cytotoxic drug that selectively destroys Leydig cells in adult testes. This study has examined the effect of a single injection of EDS on the Leydig cell populations present in the testes of rats aged 5, 10, or 20 days. Microscopic examination of the tissue demonstrated that the fetal Leydig cell population was destroyed at all ages, but that subsequent development of the adult population of Leydig cells was not affected. Whilst the destruction of the fetal Leydig cells in this acute phase of EDS on 5-day-old rats was accompanied by a decline in serum testosterone levels, there was no apparent effect on this hormone when EDS administered at 10 or 20 days of age, despite the destruction of fetal Leydig cells in these rats. The long-term effects of EDS on Day 5 of age resulted in proliferation of the intertubular tissue in which more Leydig cells were observed, but serum testosterone and testosterone levels in response to human chorionic gonadotropin stimulation in vitro were normal despite moderate or severe disruption of the seminiferous epithelium. These data show that the fetal Leydig cells of immature testes are sensitive to the cytotoxic effects of EDS in the adult, but the response of the testes differs depending on the age at which the drug is administered. PMID- 2546617 TI - Effect of adenosine triphosphate analogues on skeletal muscle fibers in rigor. AB - It is commonly believed, for both vertebrate striated and insect flight muscle, that when the ATP analogue adenyl-5'-yl imidodiphosphate (AMPPNP) is added to the muscle fiber in rigor, it causes the fiber to lengthen by 0.15%. This has been interpretated (Marston S.B., C.D. Roger, and R.T. Tregear. 1976. J. Mol. Biol. 104:263-267) as suggesting (a) that in rigor the crossbridge is fixed to, i.e., almost never detaches from the actin filament; (b), that the crossbridge remains fixed to the actin filament after AMPPNP addition; and (c) that the ability of AMPPNP to cause apparent lengthening of a muscle fiber is due to its ability to cause a conformational change in the myosin crossbridge that has an axial component of approximately 1.6 nm/half-sarcomere. The present study, done only on chemically-skinned rabbit psoas fibers, confirms that AMPPNP can cause muscle fibers to lengthen by 0.15% but only for a narrow set of experimental conditions. When experimental conditions are varied over a wider range, it becomes apparent that the extent of lengthening of a rigor muscle fiber upon AMPPNP addition depends almost entirely on the strain present in the rigor fiber before AMPPNP addition. Addition of AMPPNP to an unstrained rigor fiber (one supporting zero tension), induces zero length change while addition of AMPPNP to very highly strained rigor fibers induces length changes greater than 0.15%. The data thus do not support the hypotheses that the crossbridges remain fixed to the actin filament after AMPPNP addition and that the size of the apparent length change induced by AMPPNP is related to the size of the axial component of a conformational change. Instead, the data support the idea that the ability of AMPPNP to cause lengthening of a rigor muscle fiber is related to its ability to accelerate the rate at which strained crossbridges detach from actin and reattach in positions in lesser strain. The data do not rule out a conformational change upon AMPPNP binding, they simply make clear that any attempt to measure a force response conceivably due to a conformational change, would be more than obscured by the force changes due to crossbridges detaching and reattaching in positions of lesser strain. PMID- 2546616 TI - Cytochemical and biochemical characterization of testicular peritubular myoid cells. AB - Testicular peritubular myoid cells secrete a paracrine factor that is a potent modulator of Sertoli cell functions involved in the maintenance of spermatogenesis. These cells also play an integral role in maintaining the structural integrity of the seminiferous tubule. To better understand this important testicular cell type, studies were initiated to characterize cultured peritubular cells using biochemical and histochemical techniques. The electrophoretic pattern of radiolabeled secreted proteins was similar for primary and subcultured peritubular cells and was unique from that of Sertoli cells. Morphologic differences between Sertoli cells and peritubular cells were noted and extended with histochemical staining techniques. Desmin cytoskeletal filaments were demonstrated immunocytochemically in peritubular cells, both in culture and in tissue sections, but were not detected in Sertoli cells. Desmin is proposed to be a marker for peritubular cell differentiation as well as a marker for peritubular cell contamination in Sertoli cell cultures. Peritubular cells and Sertoli cells were also stained histochemically for the presence of alkaline phosphatase. Staining for the alkaline phosphatase enzyme was associated with peritubular cells but not with Sertoli cells. Alkaline phosphatase is therefore an additional histochemical marker for peritubular cells. Biochemical characterization of peritubular cells relied on cell-specific enzymatic activities. Creatine phosphokinase activity, a marker for contractile cells, was found to be associated with peritubular cells, while negligible activity was associated with Sertoli cells. Alkaline phosphatase activity assayed spectrophotometrically was found to be a useful biochemical marker for peritubular cell function and was utilized to determine the responsiveness of primary and subcultured cells to regulatory agents. Testosterone stimulated alkaline phosphatase activity associated with primary cultures of peritubular cells, thus supporting the observation that peritubular cells provide a site of androgen action in the testis. Retinol increased alkaline phosphatase activity in subcultured peritubular cells. Alkaline phosphatase activity increased in response to dibutyryl cyclic adenosine monophosphate (AMP) in both primary and subcultured peritubular cell cultures. Observations indicate that the ability of androgens and retinoids to regulate testicular function may be mediated, in part, through their effects on peritubular cells. This provides additional support for the proposal that the mesenchymal-epithelial cell interactions between peritubular cells and Sertoli cells are important for the maintenance and control of testicular function. Results imply that the endocrine regulation of tissue function may be mediated in part through alterations in mesenchymal-epithelial cell interactions. PMID- 2546618 TI - Receptors for erythropoietin in mouse and human erythroid cells and placenta. AB - High and lower affinity receptors for erythropoietin (EP) were initially identified on a very pure population of EP-responsive erythroblasts obtained from the spleens of mice infected with anemia strain of Friend virus (FVA). The structure of the receptor for EP in these cells was determined to be proteins of 100 and 85 Kd by cross-linking 125I-EP. In this investigation, studies on the receptors for EP were extended to other mouse erythroid cells and human erythroid cells as well as to the placentas of mice and rats. Only lower affinity receptors for EP were detected on erythroblasts purified from the spleens of mice infected with the polycythemia strain of Friend virus and a murine erythroleukemia cell line, both of which are not responsive to EP in culture. Internalization of 125I EP was observed in both groups of cells. The structure of the receptor determined by cross-linking 125I-EP was two equally labeled proteins of 100 Kd and 85 Kd molecular mass in all these mouse erythroid cells. The structure of the receptor was found to be very similar in human erythroid colony forming cells cultured from normal blood. These cells respond to EP with erythroid maturation and were previously shown to have high and lower affinity receptors. Placentas from mice and rats were found to have only lower affinity receptors for EP, and when placental membranes were cross-linked to 125I-EP, the same 100 Kd and 85 Kd bands were found as seen in mouse and human erythroid cells. The structure of the receptor was similar in cells that have high affinity receptors (FVA-infected and human erythroid colony-forming cells) and nonresponsive erythroid cells and placenta that have lower affinity receptors, but only the cells with the high affinity receptors respond to the addition of EP with erythroid maturation. PMID- 2546619 TI - Collagen-platelet interactions: evidence for a direct interaction of collagen with platelet GPIa/IIa and an indirect interaction with platelet GPIIb/IIIa mediated by adhesive proteins. AB - Using intact human platelets as the immunogen and a functional, collagen-coated bead agglutination assay, we have produced a murine monoclonal antibody (6F1) that blocks the interaction between platelets and collagen in the presence of Mg++. 6F1 affinity-purified the platelet glycoprotein Ia/IIa complex, and approximately 800 molecules of 6F1 bound per platelet at saturation. 6F1 nearly completely inhibited collagen-induced platelet aggregation and inhibited platelet adhesion to collagen by greater than 95% when plasma proteins were absent. Antibody 10E5, which blocks the binding of adhesive glycoproteins to GPIIb/IIIa, produced only minor inhibition (approximately 25%) of adhesion under the same circumstances. In contrast, when tested in platelet-rich plasma (PRP), 6F1 had only a minor effect on collagen-induced platelet aggregation, prolonging the lag phase but not the slope or maximum aggregation. Similarly, when collagen was precoated with plasma, 6F1 caused less inhibition of platelet adhesion (53%) than without the precoating (greater than 95%). Antibody 10E5 inhibited this adhesion by 32%, and the combination of 6F1 and 10E5 was more effective than either alone, inhibiting it by 90%. Time course studies of platelet agglutination of collagen coated beads using PRP containing physiologic concentrations of divalent cations showed early inhibition by 6F1, indicating that the GPIa/IIa receptor operates in this environment. With more prolonged incubation, however, 6F1 was less effective; this later agglutination could be partially prevented by adding 10E5 or PGE1 to the 6F1. These data support a model wherein collagen can directly interact with GPIa/IIa and can indirectly interact with GPIIb/IIIa via intermediary adhesive proteins. The physiological significance of these interactions, and potential interactions with other receptors, remains to be established. PMID- 2546620 TI - Clonal expansion of lymphocytes bearing the gamma delta T-cell receptor in a patient with large granular lymphocyte disorder. AB - Repeated analysis of peripheral blood lymphocytes (PBLs) from a patient with large granular lymphocytosis, neutropenia, and rheumatoid arthritis revealed that approximately 45% of PBLs displayed the following phenotype: CD3+, CD4-, CD8-, CD16+, HNK-1-, WT31-. This population was purified for further analysis by depletion with anti-CD4 and anti-CD8 monoclonal antibodies (MoAbs). Southern blot analysis showed preferential rearrangements of the V gamma 9 genes. Northern blot demonstrated the presence of V gamma 9 mRNA transcripts. With MoAbs directed against either the V gamma 9 peptide (Ti gamma A) or the delta chain of the gamma delta T-cell receptor (TCR delta 1), we further demonstrated that those cell surfaces expressed both V gamma 9 and a delta gene product. In addition, analysis of the gamma gene rearrangements on six clones derived from this population demonstrated a unique rearrangement on a single chromosome, strongly suggesting the monoclonality of this T-cell population. Significant cytotoxic activity against K562, U937 was observed only after an in vitro culture period with interleukin-2 (IL-2), whereas no specific inhibitory effect on autologous bone marrow (BM) CFU-G was noted. PMID- 2546621 TI - Antigen-specific recognition of autologous leukemia cells and allogeneic class-I MHC antigens by IL-2-activated cytotoxic T cells from a patient with acute T-cell leukemia. AB - Culturing of leukemic blood lymphocytes from a patient with acute T-cell lymphoblastic leukemia (T-ALL) with interleukin-2 (IL-2) yielded T-cell line AK-1 with a remarkable cytotoxic specificity. This line mediated strong lysis of tumor target lines expressing major histocompatibility complex (MHC) class I antigens, such as Raji, CEM, and Molt-4 cells, but no killing of K562 and Daudi cells, which are deficient in MHC class I. In contrast, lymphokine-activated killer (LAK) cells from normal donors destroyed all these tumor targets, without MHC restriction. Line AK-1, originating from residual normal T cells present in the leukemic blood, lysed autologous leukemic blasts and peripheral blood lymphocytes (PBL) from many but not all allogeneic individuals but failed to kill autologous remission lymphocytes. Destruction of the autologous leukemic targets by AK-1 could be inhibited by unlabeled competitor target cells that were lysed by AK-1, but not by target cells that were not lysed. This suggests that AK-1 specifically recognized an alien determinant on the autologous ALL cells, crossreactive with allogeneic MHC class I antigens. This reactivity with some degree of tumor specificity may be a leukemic equivalent to responses reported for populations of tumor infiltrating lymphocytes (TIL) seen in some solid tumors. PMID- 2546622 TI - Cross-lineage gene rearrangements in human leukemic B-precursor cells occur frequently with V-DJ rearrangements of IgH genes. AB - Gene configurations of immunoglobulin (Ig), T-cell antigen receptor (TCR) beta chain, and T-cell rearranging gene gamma (TRG gamma) were studied in human B precursor lymphoblastic leukemic cells. These cells were phenotypically classified into three developmental stages (stages II through IV) according to Nadler's criteria. All of them showed the Ig heavy chain (IgH) gene rearrangement, and 67% of the cells in stage IV had the rearranged TRG gamma, albeit seldom in other stages. We further analyzed IgH gene rearrangements in detail using upstream to DH (diversity region of IgH) region probe to distinguish DJ from V-DJ recombination. All dual genotyped cells in each stage except one case in stage II showed the V-DJ rearrangements. This suggests the cross-lineage involvement of the putative recombinase, particularly in the process of V-DJ rearrangements. We next examined the transcriptional status of Ig genes as an indirect reflection of the accessibility of these genes to the recombinase. Properly spliced IgH transcripts of normal size were observed in stage IV and surface Ig positive stage, but not in stage II nor III. However, IgH transcripts of aberrant sizes were seen in stage II, III, and also IV. Cross-lineage gene rearrangements were shown to occur frequently when normally spliced IgH gene begins to be transcribed or just before this. These findings may implicate that V DJ or V-VDJ gene rearrangements forming functional IgH genes, induce frequently TCR or TRG gene rearrangements. We propose these dual genotypes are different in origin from those observed in stem cell leukemia. PMID- 2546624 TI - Establishment and characterization of an Epstein-Barr virus transformed cell line with strong phagocytic activity. AB - An immunoglobulin M (IgM)-positive cell line, Ms 28, apparently spontaneously transformed by Epstein-Barr virus (EBV) was established from peripheral blood cells of a patient with immature myeloblastic leukemia. It has been characterized according to phenotype, cytochemistry, and membrane antigen pattern. The cell line expresses lymphoid markers like CD 19, CD 22, and CD 30 and synthesizes and secretes IgM. Monocyte markers CD 11c, CD 14, and CD 15 are absent. Neither interleukin-1 (IL-1), nor tumor necrosis factor (TNF-alpha) are produced. But Ms 28 cells show strong phagocytic activity and engulf Latex particles and sheep RBCs (SRBCs) that need not to be opsonized. The phagocytic activity can be inhibited by chloroquine. Both phagocytosis and EBV nuclear-antigen (EBNA) expression can be observed in one and the same cell. Ms 28 cells might be useful to study immunologic activities like antigen processing and presentation. PMID- 2546623 TI - Establishment and characterization of a childhood T-cell acute lymphoblastic leukemia cell line, PER-255, with chromosome abnormalities involving 7q32-34 in association with T-cell receptor-beta gene rearrangement. AB - A human leukemia cell line, PER-255, was established from the bone marrow of a 5 year-old boy with features typical of lymphomatous T-acute lymphoblastic leukemia (T-ALL). The leukemic origin of cell line PER-255 is indicated by its cytochemical and immunologic similarity to the patient's fresh leukemic cells, which correspond to immature cortical thymocytes. Southern blot analysis showed that the IgJH genes were in germline configuration, whereas both alleles of the T cell receptor-beta (TCR-beta) gene were rearranged in PER-255 cells, with identical rearrangements present in the patient's leukemic cells. Cytogenetic analysis of the cell line revealed a single abnormal clone with the karyotype 46,XY,t(7;10)(q32-34;q24),t(9;12) (p22;p12-13). Reciprocal translocations involving chromosome bands 7q32-36, containing the gene for the TCR-beta chain, have been reported for a number of tumors of T-cell origin. Translocations involving the 7q32-36 region appear to be nonrandomly associated with childhood T ALL, whereas abnormalities of 9p and 12p have been reported to be nonrandomly involved in ALL but not specifically associated with the T-cell phenotype. PMID- 2546625 TI - Feline parvovirus propagates in cat bone marrow cultures and inhibits hematopoietic colony formation in vitro. AB - Feline parvovirus (FPV) causes leukopenia in naturally infected cats. We investigated the mechanism of hematopoietic depression by this virus in feline bone marrow cultured in vitro. In suspension cultures we demonstrated FPV propagation and replication using DNA molecular hybridization. Viral RNA and DNA were observed by in situ hybridization in about 10% of marrow cells at day 3. Granulocytes and their precursors were virtually absent from infected cultures after six days. Infected cells showed viral capsid protein predominantly in nuclei by immunofluorescence. In clonal assays, FPV most efficiently inhibited hematopoietic colony formation by myeloid progenitor cells (CFU-GM), but erythroid colony formation (BFU-E and CFU-E-derived) was also depressed in the presence of virus. Inhibition of colony formation could be abrogated by physical inactivation of the virus or preincubation with specific neutralizing antibodies. Recombinant human colony stimulating factors GM-CSF and G-CSF supported feline myelopoiesis in progenitor assays, and FPV completely inhibited factor dependent colony formation. PMID- 2546626 TI - Expression of retinoic acid receptor alpha mRNA in human leukemia cells. AB - The expression of the newly described human retinoic acid receptor alpha (RAR alpha) in six nonlymphoid and six lymphoid leukemia cell lines and nine freshly obtained samples of leukemia cells from patients with acute nonlymphoid leukemia was assessed by Northern blot analysis, using a full length cDNA clone of RAR alpha as probe. RAR alpha was expressed in all 12 cell lines and in all fresh leukemia samples as two major transcripts of 2.6 and 3.5 kb in size. Levels of RAR alpha expression and transcript sizes in retinoid-sensitive cells (such as HL60 or fresh promyelocytic leukemia cells) were not different from those in other samples. Moreover, expression of RAR alpha was not significantly modulated by exposure to cis-retinoic acid (cisRA) in either cisRA-responsive or unresponsive cells. By using a 3' fragment of the RAR alpha gene as a probe, we confirmed that the transcripts visualized did not represent the homologous RAR beta gene. RAR alpha appears to be expressed in most human leukemia cells regardless of the type of biologic response to retinoic acid. PMID- 2546627 TI - Interleukin-3 and granulocyte-monocyte colony-stimulating factor receptors on human acute myelocytic leukemia cells and relationship to the proliferative response. AB - Interleukin-3 (IL-3) and granulocyte-monocyte-colony-stimulating factor (GM-CSF) stimulate proliferation of human acute myeloid leukemia (AML) in vitro, although patterns of response among clinical cases are diverse. Whether regulatory abnormalities related to growth factor responses in human AML may establish the outgrowth of the neoplasm is unclear. We determined receptor numbers and affinity for IL-3 and GM-CSF on human AML cells using human recombinant IL-3 (rIL-3) and GM-CSF (rGM-CSF). In 13 of 15 cases of primary AML high-affinity (kd 26 to 414 pmol/L) receptors for IL-3 were demonstrable on the cells. The average numbers of IL-3 receptors ranged from 21 to 145 receptors per cell. Normal WBCs showed IL-3 receptors on their surface at similar densities. IL-3 receptor positivity often correlated with GM-CSF receptor positivity of AML; GM-CSF receptors were demonstrated on the cells of 11 of 15 cases, although average numbers of GM-CSF receptors were ten times greater. The in vitro response of the cells to exogenous IL-3 or GM-CSF was examined by measuring thymidine uptake. Because IL-3 and GM CSF were potent inducers of DNA synthesis in vitro, apparently relatively few receptors are required to permit activation of growth. These experiments did not provide evidence for overexpression or increased receptor sensitivity as an explanation for AML growth. In a minority of cases, however, the cells were unable to respond to IL-3 (four of 15 cases) or GM-CSF (four of 15 cases) despite normal receptor availability on the cell surface. PMID- 2546628 TI - Induction of dependence on hematopoietic proteins for viability and receptor upregulation in differentiating myeloid leukemic cells. AB - There are different types of myeloid leukemic cells that can be induced to differentiate to mature granulocytes or macrophages by different hematopoietic regulatory proteins. One type of leukemic clone can be induced to differentiate by recombinant macrophage and granulocyte differentiation-inducing protein-type 2 (MGI-2), which we have shown is Interleukin-6 (IL-6), and another type of leukemic clone can be differentiated by recombinant granulocyte-macrophage colony stimulating factor (GM-CSF) or IL-3. There was no subpopulation of growth factor responsive or differentiation-defective cells before induction of differentiation in either type of clone. In both clones, induction of differentiation-induced requirement for a hematopoietic protein for cell viability. Viability of the cells was maintained by IL-6, IL-3, or macrophage colony-stimulating factor (M CSF) but not by GM-CSF in the cells differentiated by IL-6, and by GM-CSF or IL-3 but not by IL-6 or M-CSF in the cells differentiated by GM-CSF or IL-3. The viable cells with a differentiated phenotype continued to multiply. In undifferentiated leukemic cells with no or few surface receptors for some of these proteins, there was an upregulation of the number of receptors during differentiation for the proteins to which the cells responded. But there were also differentiating leukemic cells with an upregulation of GM-CSF receptors although GM-CSF could not maintain the viability of the differentiating cells. The results indicate that induction of hormone responsiveness and upregulation of the hormone receptors can both occur in differentiating leukemic cells, and that the regulation of these two events can be separated. PMID- 2546629 TI - Transcellular sulfidopeptide leukotriene biosynthetic capacity of vascular cells. AB - Cells in the vasculature, including polymorphonuclear leukocytes, platelets, and endothelial cells, have been shown to be jointly involved in the biosynthesis of active lipid mediators derived from arachidonic acid. Stimulation of neutrophils with the calcium ionophore A23187 as a model for cell activation results in production of leukotriene (LT)A4 with subsequent intracellular conversion into LTB4. When platelets or endothelial cells were present in the incubation system, LTC4 was produced from the neutrophil-derived LTA4. Whereas production and release of LTA4 under resting conditions in vivo might be expected to be quite low, under pathologic conditions, LTA4 production could be markedly increased. Therefore, the metabolism of exogenous LTA4 by platelets and endothelial cells was studied at a wide range of LTA4 concentrations. The production of LTC4 during coincubation of neutrophils with platelets was found to be dependent on neutrophil number ranging from 2 x 10(5) to 2 x 10(7) cells/mL. When a fixed number of neutrophils were stimulated with platelets alone or with mixtures of platelets and endothelial cells, LTC4 production was observed to be dependent on both acceptor cell types. These results suggest that mixed cell populations, which are likely to occur in vivo, may be critical determinants of the profile of eicosanoids produced in pathophysiologic circumstances. We suggest that both endothelial cells and platelets, in the presence of neutrophils, contribute large quantities of sulfidopeptide leukotrienes to inflammatory and thrombotic situations. Furthermore, platelets, because of their quantity and reactivity, may play a pivotal role in transcellular biosynthesis of eicosanoids. PMID- 2546630 TI - Montreal platelet syndrome: a defect in calcium-activated neutral proteinase (calpain). AB - Platelets from patients with Montreal platelet syndrome (MPS) consistently display a defect in the mechanisms that regulate platelet size during shape change and undergo spontaneous aggregation and stir-induced microaggregate formation. We now provide data that the surface glycoprotein composition of MPS platelets is indistinguishable from that of normal platelets. However, a defect in calcium-activated neutral proteinase (calpain) was detected in MPS platelets. The specific activity of calpain in the cytosolic fraction of platelets from four MPS patients was found to be only 30% of that in platelets from normal control donors (n = 18, P less than .001). Additionally, platelets from MPS patients (n = 3) contained only 50% (P less than .001) of the calpain I catalytic subunit antigen found in platelets from normal control donors (n = 9). Platelets from the asymptomatic father/grandfather of the MPS patients had normal amounts of both total calpain proteolytic activity and calpain I catalytic subunit antigen. This represents the first report of a defect in calpain in human cells. The abnormally low calpain activity in MPS platelets may account for the platelet defects characteristic of this disorder. PMID- 2546631 TI - Plasminogen binding to rat hepatocytes in primary culture and to thin slices of rat liver. AB - Human 125I-plasminogen bound readily to rat hepatocytes in primary culture at 4 degrees C and at 37 degrees C. Binding was inhibited by lysine and reversed by lysine, epsilon-aminocaproic acid, or nonradiolabeled plasminogen. The Kd for binding of 125I-plasminogen to hepatocytes was 0.59 +/- 0.16 mumol/L, as determined from the saturation isotherm by nonlinear regression (r2 = 0.99) and the Scatchard transformation by linear regression (r2 = 0.93). The number of sites per cell was 14.1 +/- 1.1 x 10(6). Fibrinogen synthesis and secretion by hepatocytes was insufficient to account for the major fraction of plasminogen binding, as determined by enzyme-linked immunosorbent assay (ELISA). Polyacrylamide gel electrophoresis and trichloroacetic acid precipitation studies demonstrated that plasminogen is neither activated nor degraded when bound to hepatocytes at 37 degrees C. Thin slices of whole rat liver (500 microns), isolated and prepared totally at 4 degrees C, bound 125I-plasminogen. Binding was inhibited by lysine. 125I-albumin binding to liver slices was minimal and not inhibited by lysine. Activation of plasminogen by tissue plasminogen activator (t PA) was enhanced by hepatocytes in primary culture. When lysine was included in the media, the enhanced rate of activation was no longer observed. After activation with t-PA, much of the plasmin remained associated with hepatocyte surfaces and was partially protected from inhibition by alpha 2-antiplasmin. These studies suggest that hepatocyte plasminogen binding sites may provide important surface anticoagulant activity. PMID- 2546632 TI - Human neutrophil-mediated lysis of ovarian cancer cells. AB - Because of recent questions concerning the sensitivity of human tumor cells to neutrophil-induced oxidative injury, we studied six freshly obtained human ovarian cancer (OC) specimens. Stimulation of neutrophils (PMNs) by phorbol myristate acetate (PMA) did not result in OC cytolysis during the first nine hours of incubation. However, three of six specimens were significantly lysed by stimulated PMNs when assay length was increased to 18 hours. Cytotoxicity was mediated by PMN production of reactive oxidative intermediates (ROIs). Presentation of ROIs to OC targets as preformed or enzymatically generated molecules in cell-free systems duplicated the enhanced lysis at 18 hours (as compared with six hours). Since addition of catalase at three or six hours did not inhibit enhanced lysis at 18 hours (achieved by PMNs or in cell-free systems), it appears that an initial ROI-mediated lethal event occurs early, but longer incubations are required for the event to become manifested as cell death. These data suggest that shorter assays may underestimate the potential of PMNs as effector cells against human tumor cells. PMID- 2546633 TI - Demonstration of monoclonal EBV genomes in Hodgkin's disease and Ki-1-positive anaplastic large cell lymphoma by combined Southern blot and in situ hybridization. AB - Forty-two cases of Hodgkin's disease (HD) and 22 cases of Ki-1-positive anaplastic large cell (Ki-1 + ALC) lymphoma were examined by Southern blotting for the presence of Epstein-Barr virus (EBV) DNA. Seven cases of HD and one case of Ki-1 + ALC lymphoma scored positive with a probe specific for the internal repetitive region of the EBV genome. Subsequently, these viral genomes could be demonstrated to be monoclonal in origin using an EBV terminal region DNA probe. In situ hybridization revealed that in two HD cases, the EBV infected cells had the distinct morphology of Hodgkin and Reed-Sternberg cells, thus suggesting a direct pathoetiological relationship between EBV and some cases of HD. PMID- 2546634 TI - T-cell receptor delta/alpha rearrangements in lymphoid neoplasms. AB - Rearrangements within the T-cell receptor (TCR)delta/alpha locus were analyzed in a wide variety of lymphoid neoplasms by eight DNA probes specific for TCR J delta, J alpha and C alpha segments. In all 11 T-cell malignancies, rearrangement and/or deletion of TCR delta was detected irrespective of the stage of maturation of the tumor. The organization of TCR delta correlated with the phenotype of the tumor: In "prethymic" T-cell acute lymphocytic leukemia (ALL), TCR delta was the only TCR gene to be rearranged. More mature T cell malignancies expressing CD4 together with CD3 showed deletion of both alleles of TCR delta, suggestive of TCR V alpha-J alpha rearrangement. All 43 B-cell tumors expressing surface immunoglobulin (sIg), including two cases of adult B-cell ALL, had germline configuration of TCR delta/alpha. In contrast, all 17 B-cell precursor ALLs (null, common, and pre-B-cell ALLs) had rearrangement and/or deletion of TCR delta/alpha. A single case of "histiocytic" lymphoma also showed biallelic deletion of TCR delta. Oligoclonal rearrangements of Ig and TCR genes were observed in two cases of B-cell precursor ALL and in one case of T-cell lymphoblastic lymphoma. Patterns of such "aberrant" TCR rearrangement were similar to those observed in T-lineage malignancies. In particular, seven of eight cases of B-cell precursor ALL and the histiocytic lymphoma which demonstrated biallelic TCR delta deletion, (suggestive of a V alpha-J alpha rearrangement) had clonal TCR beta rearrangement. These data support the hypothesis that supposedly aberrant rearrangements of the TCR genes may follow the same developmental controls as found in T-cell differentiation, despite the lack of evidence for further commitment to the T-cell lineage. TCR delta rearrangement is a useful marker of clonality of immature T-cell tumors which may have only this gene rearranged but is not specific to the T-cell lineage. PMID- 2546635 TI - Biotinylated recombinant human erythropoietins: bioactivity and utility as receptor ligand. AB - Recombinant human erythropoietin labeled covalently with biotin at sialic acid moieties has been prepared, and has been shown to possess high biological activity plus utility as a receptor ligand. Initially, the effects on biological activity of covalently attaching biotin to erythropoietin alternatively at carboxylate, amino, or sialic acid groups were compared. Biotinylation of erythropoietin at carboxylate groups using biotin-amidocaproyl hydrazide plus 1 ethyl-3-(3-dimethylaminopropyl) carbodiimide led to substantial biological inactivation, although biotinylated molecules retained detectable activity when prepared at low stoichiometries. Biotinylation at amino groups using sulfosuccinimidyl 6-(biotinamido) hexanoate resulted in a high level of biological inactivation with little, if any, retention of biological activity, regardless of labeling stoichiometries. Biotinylation at sialic acid moieties using periodate and biotinamidocaproyl hydrazide proceeded efficiently (greater than 95% and 80% labeling efficiencies for human urinary and recombinant erythropoietin, respectively) and yielded stably biotinylated erythropoietin molecules possessing comparably high biological activity (ie, 45% of the activity of unmodified hormone). Utility of recombinant biotin-(sialyl)-erythropoietin (in combination with 125I-streptavidin) in the assay of cell surface receptors was demonstrated using two distinct murine erythroleukemia cell lines, Friend 745 and Rauscher Red 1. The densities and affinities of specific hormone binding sites were 116 +/- 4 sites, 3.3 +/- 0.4 nmol/L kd and 164 +/- 5 sites, 2.7 +/- 0.4 nmol/L kd, respectively. It is predicted that the present development of biotin (sialyl)-erythropoietin as a chemically and biologically stable, bioactive ligand will assist in advancing an understanding of the regulated expression and physicochemistry of the human and murine erythropoietin receptors. PMID- 2546636 TI - Macrophage colony-stimulating factor (CSF-1) is expressed by spontaneously outgrown EBV-B cell lines and activated normal B lymphocytes. AB - Human B lymphocytes activated by mitogens or infected by Epstein Barr virus (EBV) have previously been shown to release colony-stimulating activity (CSA) supporting the growth of normal human bone marrow progenitors. We established five different human EBV-B cell lines spontaneously outgrown from nonmalignant peripheral blood cells and long-term bone marrow cultures. CSA derived from all of these lines induces the growth of murine macrophage colonies, whereas virtually no human bone marrow cell progenitors were stimulated. As observed in the tumor cell line MIA PaCa-2, a 4.3-kilobase (kb) transcript was detected in all cases using a human colony-stimulating factor (CSF)-1 probe. Expression of this transcript can be further stimulated within three hours upon addition of phorbol myristate acetate (PMA). The highly purified native protein exerting macrophage colony-stimulating activity (M-CSA) exhibits a molecular size of approximately 75 to 97 Kd in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The identity of EBV-B cell derived M-CSA with human urinary CSF-1 was confirmed by a complete neutralization of macrophage CSA by an antihuman urinary CSF-1 antiserum. Normal human B lymphocytes purified from tonsils or from mononuclear blood cells also express CSF-1 upon stimulation with Staphylococcus aureus Cowan I. No CSF-1 expression, however, could be detected in normal resting B lymphocytes or in the Burkitt lymphoma cell line RAJI. PMID- 2546637 TI - Modulation of platelet function by extracellular adenosine triphosphate. AB - A potential physiologic role of extracellular adenosine triphosphate (ATP) on platelet function is proposed in this report. It is widely accepted that ATP competitively inhibits adenosine diphosphate (ADP)-induced platelet aggregation. Our observations of platelet aggregation with the agonists, collagen, epinephrine, and ADP in the presence of 180 mumol/L ATP could support this competitive nature of ATP. However, the disaggregation of maximally aggregated platelets induced by ATP, theophylline, or ATP plus theophylline indicates that additional mechanisms of ATP action may be present. Extracellular gamma-32P-ATP (7 pmol) labels surface-membrane proteins in intact platelets as demonstrated by several criteria. The reaction is Ca++-dependent. Stimulation by calcium occurs in the physiologic range of 1 to 5 mmol/L. Significant levels of phosphorylation occur within one minute with near maximal levels reached by five minutes. Platelet cyclic AMP (cAMP) levels were elevated in a dose-dependent fashion in cells incubated for four minutes with increasing amounts of extracellular ATP (18 to 540 nmol). The addition of ATP plus theophylline resulted in a synergistic stimulation of cAMP levels. ATP was not being hydrolyzed to adenosine by plasma nucleotidases, as demonstrated by the lack of effect of ten U of adenosine deaminase. The phosphorylation of surface proteins by extracellular ATP released from activated platelets may modulate platelet responsiveness to agonists at distances removed from the site of vascular injury. Phosphorylation may also play a role in signal transduction to regulate the levels of intracellular cAMP, which further inhibits platelet activation. PMID- 2546638 TI - Effects of human PTH-related peptide and human PTH on cyclic AMP production and cytosolic free calcium in an osteoblastic cell clone. AB - Recently, human parathyroid hormone-related peptide (hPTHrP) has been purified and its amino acid sequence determined. Within the amino-terminal 13 residues of hPTHrP, 8 amino acids were found homologous between hPTHrP and human PTH (hPTH). This peptide was reported to stimulate cyclic AMP (cAMP) production in osteoblastic cell lines (UMR106 and ROS17/2.8). However, whether or not this peptide affects another second messenger, i.e., cytosolic free calcium ([Ca2+]i), in osteoblasts has not yet been determined. Therefore, in the present study, we examined the effects of synthetic amino-terminal fragments of hPTHrP (Tyr40hPTHrP1-40 and hPTHrP1-34) on intracellular cAMP production and [Ca2+]i in an osteoblastic cell line (MC3T3-E1) and compared them with those of hPTH1-34. Human PTHrP1-34, Tyr40hPTHrP1-40 and hPTH1-34 stimulated cAMP production in an equipotent manner at concentrations ranging from 2.5 x 10(-10) to 1.3 x 10(-6) M. Human PTH1-34 at concentrations from 2.5 x 10(-7) to 1.3 x 10(-6) M significantly (P less than 0.05) increased [Ca2+]i, but hPTHrP1-34 and Tyr40hPTHrP1-40 at the same concentrations did not. These results suggest a different receptor-mediated mechanism for [Ca2+]i increase between hPTHrP and hPTH, although these two peptides appear to share the same receptor site(s) which is coupled to the cAMP system in MC3T3-E1. PMID- 2546639 TI - Corticosteroid-induced changes in the responsiveness of human osteoblast-like cells to parathyroid hormone. AB - We investigated the in vitro effect of corticosteroids on the responsiveness of human cells of osteoblast lineage to parathyroid hormone (PTH). Prior to corticosteroid treatment, the cells demonstrated only a small increase in cAMP production and no measurable change in transmembrane potential in response to PTH. Exposure of cells to dexamethasone resulted in a 5-fold increase in PTH induced cAMP production and in measurable PTH-induced membrane depolarization in all cells studied. The effect of corticosteroids on cAMP production was specific for PTH (not seen with PGE1 or forskolin), occurred in a time- and dose-dependent fashion and in the absence of cell proliferation. Most of the cells were of osteoblast lineage as determined by the presence of alkaline phosphatase activity and BGP secretion. These findings further support the idea that corticosteroids increase the sensitivity of cells of osteoblast lineage to PTH, perhaps by transforming cells which initially have a low responsiveness to PTH to a state of high responsiveness. PMID- 2546640 TI - Avoiding injuries caused by pigs. PMID- 2546641 TI - Tridimensional analysis of the formation of secretory vesicles in the Golgi apparatus of absorptive columnar cells of the mouse colon. AB - The tridimensional structure of the Golgi apparatus has been studied in the absorptive cells of the mouse colon by means of reduced osmium postfixation and phosphatase cytochemistry. In thick sections of tissue impregnated with osmium tetroxide or treated with a technique to demonstrate TPPase activity, the Golgi formed a continuous ribbon-like structure capping the upper pole of the nucleus. Along the longitudinal axis of this ribbon, compact zones made up of superposed flattened saccules alternated with less compact zones which consisted of highly perforated saccules or bridging anastomosed tubules. In the cis-trans axis, the following elements were observed: (1) a cis element consisting of a continuous osmiophilic tubular network; (2) two or three subjacent elements selectively perforated by wells; (3) a trans compartment made up of two or three TPPase reactive sacculotubular elements, some showing a "peeling-off" configuration. In some regions, the first flattened saccule of this trans compartment displayed discrete ovoid dilatations, located in compact zones and containing a dense granulofibrillar material; in the subjacent elements this material was seen concentrated in nodular swellings, at the intersection of the meshes of anastomosed membranous tubules. 100-300 nm vesicles containing a similar dense granulofilamentous material were observed in the trans Golgi zone and interspersed in the supranuclear cytoplasm between the Golgi zone and the apical surface of the cell. Smaller vesicles 80-100 nm in diameter containing a fine dusty material were also seen in proximity. These morphological observations suggested that at least two kinds of material were segregated in the saccules of the trans compartment and packaged in vesicles of two class sizes that detached from the Golgi stack on its trans aspect. PMID- 2546642 TI - Carcinoma of the prostate. AB - In Scotland, carcinoma of the prostate is now the third most common malignancy in males. The disease is only curable if removed or irradiated when confined to the prostate but has usually spread beyond the prostate when the diagnosis is made. This article reviews the clinical presentation, diagnostic and staging methods and current treatment available for this common clinical problem. PMID- 2546643 TI - Radical retroperitoneal node dissection after chemotherapy for testicular tumours. AB - A group of 29 patients underwent salvage node dissection for residual retroperitoneal masses following orchiectomy and combination chemotherapy for advanced testicular cancer between 1980 and 1988. The results confirm that surgery for residual retroperitoneal masses following combination chemotherapy is worthwhile and whenever possible the sympathetic chain should be preserved. PMID- 2546644 TI - Adenocarcinoma in a substitution caecocystoplasty. PMID- 2546646 TI - Beta-estradiol induced catecholamine-sensitive hyperalgesia: a contribution to pain in Raynaud's phenomenon. AB - The physiological basis of the pain and hyperalgesia observed in patients with Raynaud's phenomenon (RP) is unknown. Since estrogen-induced effects on sympathetic postganglionic neurons (SPGNs) have been implicated in the vasomotor abnormalities in patients with RP, we have studied the effects of estradiol on nociceptive thresholds and noradrenaline sensitivity in a nociceptive flexion reflex in the rat. We report that estradiol induces a catecholamine sensitive hyperalgesia. This hyperalgesia is antagonized by yohimbine (an alpha 2 adrenergic antagonist) but not prazosin (an alpha 1-adrenergic antagonist) as well as by inhibitors of the cyclooxygenase pathway of arachidonic acid metabolism. These data are compatible with the hypothesis that the sensory abnormalities observed in patients with RP may depend on estradiol-induced changes in SPGN, resulting in a sympathetically-dependent production of cyclooxygenase products of arachidonic acid. PMID- 2546645 TI - Substantia nigra as a site of origin of dopamine-dependent motor syndromes induced by stimulation of mu and delta opioid receptors. AB - Opioid agonists having different affinity for delta and mu receptors were injected bilaterally in the substantia nigra (SN) of rats. The selective agonist of mu receptors N-MePhe3,-D-Pro4 morphiceptin (PLO 17) produced a stereotyped behavior characterized by stereotyped sniffing and gnawing antagonized by the irreversible antagonist of mu receptors beta-funaltrexamine. In contrast, bilateral intranigral injection of the selective delta agonist D-Pen2,D-Pen5 enkephalin (DPDPE) elicited dose-dependent exploratory behavior and rearing but failed to produce gnawing. The behavioral syndrome induced by DPDPE was significantly reduced by the selective delta antagonist ICI 174,864. Naloxine, a non-selective opioid antagonist, antagonized the effects of both compounds. SCH 23390 and haloperidol, two antagonists of dopaminergic D1 and D2 receptors, respectively, blocked the effects of PLO 17 and DPDPE. The results indicate that stimulation of specific opioid receptor types in the SN elicits specific behavioral syndromes and suggest that the SN might be the site of origin of certain items of the behavioral syndrome evoked by systemic opiates. These items might be mediated by activation of dopaminergic neurons of the ventral mesencephalon. PMID- 2546647 TI - Regulation of high-affinity GABAA receptors in the dorsal hippocampus by estradiol and progesterone. AB - The effects of estradiol benzoate (EB) and of progesterone (P) treatment on high affinity [3H]muscimol binding in the dorsal hippocampal formation were examined in ovariectomized and adrenalectomized female rats by in vitro autoradiography. EB injected subcutaneously increased [3H]muscimol binding in specific subregions of the Ammon's horn (CA1 and stratum radiatum of CA4) and of the dentate gyrus (dorsal molecular layer). In these particular regions, estrogen receptors have been shown to be present. P did not significantly affect [3H]muscimol binding in any region of the hippocampus when administered alone or in combination with EB. Results suggest that estrogens may regulate the activity of specific hippocampal neurons by modulating their sensitivity to GABA. PMID- 2546648 TI - Hypothermia enhances protective effect of MK-801 against hypoxic/ischemic brain damage in infant rats. AB - Accumulating evidence suggests that the N-methyl-D-aspartate (NMDA) subtype of glutamate receptor may play an important role in hypoxic/ischemic (H/I) brain damage. Accordingly, it has been shown that the NMDA antagonist, MK-801, partially protects the infant rat brain against H/I damage. Here we show that reducing the body temperature of the infant rat also confers partial protection against H/I brain damage and that mild hypothermia plus MK-801 treatment provides total protection against such damage. Relevance of these findings to the prevention of perinatal brain damage in humans is discussed. PMID- 2546649 TI - Scorpion toxin prolongs an inactivation phase of the voltage-dependent sodium current in rat isolated single hippocampal neurons. AB - The effects of scorpion toxin on the voltage-dependent sodium current (INa) of CA1 pyramidal neurons isolated from rat hippocampus were studied under the single electrode voltage-clamp condition using a 'concentration-clamp' technique. The toxin increased the peak amplitude of INa and prolonged its inactivation phase in a time- and dose-dependent manner. Inactivation phase of INa proceeded with two exponential components in the absence (control) and presence of the toxin. In the toxin-treated neurons, both the time constant of slow component and its fractional contribution to the total current increased dose-dependently while the fractional contribution of the fast one decreased in a dose-dependent fashion without changing its time constant. Actions of scorpion toxin on the sodium channels of hippocampal pyramidal neurons were essentially similar to those of peripheral preparations. Therefore, it can be concluded that the sodium channels of mammalian brain neurons have structures and functions similar to peripheral channels. PMID- 2546651 TI - Co-administration of Ro 15-1788 prevents diazepam-induced retardation of recovery of function. AB - Following unilateral lesions of the anteromedial cortex, recovery from somatosensory asymmetry reliably occurs within about 10 days. Chronic exposure to diazepam significantly delays this recovery. In the present study, co administration of Ro 15-1788, a benzodiazepine antagonist (i.e. it blocks the negative and positive allosteric modulation of GABA), prevented diazepam-induced retardation of recovery from somatosensory asymmetry. Nocturnal ambulatory (motor) activity was not different between rats receiving diazepam-alone and those receiving Ro 15-1788 in combination with diazepam. These data suggest that the benzodiazepine receptor is importantly involved in the detrimental effects of diazepam on recovery, and that non-specific behavior sedation plays little or no role. PMID- 2546652 TI - Chemical kindling induced by cAMP and transfer to electrical kindling. AB - Repeated injection of an initially subconvulsive dose of cAMP into the rat amygdala (AM) produced progressive seizure development similar to that of electrical kindling. The chemical kindling occurred in a dose-dependent manner. Furthermore, when cAMP was combined with EDTA, a strong inhibitor of cAMP phosphodiesterase, the seizure development was remarkably facilitated. When the animals that had been chemically kindled were subjected to electrical kindling, they rapidly developed seizures. These results, together with those previously published, suggest that cAMP participates in the seizure development induced by AM kindling. PMID- 2546650 TI - Stress-induced behavioral depression in the rat is associated with a decrease in GABA receptor-mediated chloride ion flux and brain benzodiazepine receptor occupancy. AB - Rats exposed to inescapable tailshock exhibit deficits in learning a simple shuttlebox escape task 24 h later. This syndrome has been termed 'behavioral depression' or 'learned helplessness', and is a model of stress-induced depression. In the present study a significant (25%) decrease in GABA receptor mediated chloride ion flux as measured by muscimol-stimulated 36Cl- uptake in synaptoneurosomes was found in the cerebral cortices of rats that failed the shuttlebox task as compared to naive control rats. Rats which were exposed to tailshock and subsequently learned the escape task did not show a significant difference in muscimol-stimulated 36Cl- uptake as compared to naive control rats. Similarly, rats that failed to learn the shuttlebox escape task had significantly lower in vivo [3H]Ro15-1788 specific binding in cerebral cortex (43%), hippocampus (35%) and striatum (33%) as compared to naive control rats. In cerebellum and hypothalamus, there were significant reductions in specific [3H]Ro15-1788 binding in both animals that failed and animals that learned the shuttlebox escape task as compared to naive controls. To control the stress of the footshock associated with the shuttlebox escape task, we investigated the effect of gridshock in which total footshock received was equivalent to that received by rats who failed the shuttlebox task. There were no differences in muscimol-stimulated 36Cl- uptake or in vivo [3H]Ro15-1788 specific binding between naive controls and rats administered footshock independent of a learning task. These data suggest that the development of stress-induced behavioral depression may be associated with a decrease in GABA receptor-mediated chloride channel function. PMID- 2546653 TI - Cytochemical demonstration of TPPase in myelinated fibers in the central and peripheral nervous system of the rat. AB - Thiamine pyrophosphatase (TPPase) activity was demonstrated by means of cytochemistry and electron microscopy in association with myelinated fibers in the central and peripheral nervous system of the rat. The areas studied included corpus callosum, hippocampus, cerebral cortex, cervical spinal cord and sciatic nerves. In the myelin sheaths, the enzymatic activity was found in 3 locations: (1) within oligodendroglial and Schwann cytoplasmic clefts between myelin lamellae; (2) in the major dense line of myelin; and (3) within the periaxonal space. In addition to this myelin-associated TPPase, enzymatic activity was also observed in specific cytoplasmic localizations in myelinogenic cells. Oligodendrocytes displayed TPPase activity within the nuclear envelope and the endoplasmic reticulum cisternae, whereas Schwann cells displayed TPPase activity within the endoplasmic reticulum and Golgi saccules. The results are discussed in relation to the role that TPPase might play in myelinated fibers, including roles in the conduction of nerve impulses or roles in the maintenance of structural configuration of myelin sheaths. PMID- 2546654 TI - Human retinal pigment epithelial cells in culture possess A2-adenosine receptors. AB - Adenosine agonists cause a marked stimulation in cyclic AMP accumulation in whole human retinal pigment epithelial (RPE) cells in the presence of adenosine deaminase and papaverine, a phosphodiesterase inhibitor. N Ethylcarboxamidoadenosine (NECA) stimulates cyclic AMP accumulation 16.1-fold above basal with an EC50 of 2.5 x 10(-7) M. It is also an effective (1.9-fold) stimulator of adenylate cyclase activity in RPE membrane preparations and a modest (1.22-fold) stimulator in the presence of forskolin in RPE cell membranes prepared from freshly isolated porcine RPE. N6-Cyclopentyladenosine (CPA) and N6 phenylisopropyladenosine (PIA) also increase cyclic AMP levels with EC50s of 4.9 x 10(6) M (8.9-fold above basal) and 3.5 x 10(-6) M (8.0-fold above basal) respectively. This potency order (NECA greater than PIA greater than CPA) is typical of A2-adenosine receptors. The relatively A1-selective agonists 10(-7) M indicating that RPE cells do not have A1-receptors which inhibit adenylate cyclase. Three adenosine receptor antagonists, BW-A1433U, 8 cyclopentyltheophylline and 8-sulfophenyltheophylline, blocked the NECA-induced stimulation of cyclic AMP accumulation with IC50s of 0.36 microM, 1.5 microM, and 75 microM respectively. Since alteration of cAMP levels has been demonstrated to affect several RPE functions, including cell migration, resorption of subretinal fluid, and phagocytosis, adenosine may play a significant regulatory role in RPE. PMID- 2546655 TI - The relationship of benzodiazepine binding sites to the norepinephrine projection in the hypothalamus of the adult rat. AB - The relationship of benzodiazepine binding sites to noradrenergic terminals in different brain regions of adult rats was evaluated. The in vitro addition of 10( 5) M diazepam (DZ) significantly reduced the depolarization-induced release of [3H]norepinephrine (NE) in the hypothalamus, while no effect was observed in the cerebellum, suggesting that DZ may act directly on NE terminals in the hypothalamus. The action of DZ in the hypothalamus was blocked either by co incubation with the benzodiazepine (BZ) antagonist RO 15-1788 (a neutral ligand for BZ binding sites) or with the GABA antagonist, bicuculline. Analysis of BZ binding heterogeneity, using the triazolopyridazine CL 218,872 as displacing agent, indicated that binding heterogeneity exists in both the cortex and hypothalamus, whereas the cerebellum has a more homogeneous binding site population. Binding site heterogeneity does not appear to explain the selective effect of DZ on hypothalamic noradrenergic terminals. Systemic injection of the neurotoxin 6-hydroxyDOPA decreased NE levels significantly in the cerebellum, cortex and hypothalamus. Only in the hypothalamus, however, did the decrease in NE levels correlate in magnitude with the decrease in BZ receptor binding. A large proportion of BZ receptors may be located presynaptically on the hypothalamic NE terminals. The location of these BZ binding sites may contribute to the unique response to DZ observed in the hypothalamus following either in vitro or in utero exposure. PMID- 2546656 TI - Ischemia triggers NMDA receptor-linked cytoskeletal proteolysis in hippocampus. AB - Transient forebrain ischemia is followed within minutes by accelerated proteolysis of the cytoskeletal protein, spectrin. This effect is most pronounced in the selectively vulnerable CA1 region of hippocampus which also experiences a second proteolytic phase during the terminal stages of neuronal degeneration. Both proteolytic phases are suppressed by MK-801, an NMDA receptor antagonist. Cytoskeletal disruption, via NMDA receptor-linked proteolytic events, is suggested to predispose vulnerable neurons to delayed cell death. PMID- 2546657 TI - Pyrethroids: involvement of sodium channels in effects on inositol phosphate formation in guinea pig synaptoneurosomes. AB - The effects of pyrethroids were studied on phosphoinositide breakdown in guinea pig synaptoneurosomes. Similar to other agents that activate voltage-dependent sodium channels, type I and type II pyrethroids stimulated phosphoinositide breakdown. Type II pyrethroids, like deltamethrin and fenvalerate, were more potent and, at least for deltamethrin, more efficacious than type I pyrethroids, like allethrin, resmethrin and permethrin. The effects of type II pyrethroids could be partially inhibited by the sodium channel blocker tetrodotoxin. The effects of allethrin and resmethrin were not affected by 5 microM tetrodotoxin. Stimulation of phosphoinositide breakdown by fenvalerate was additive to the stimulation elicited by the receptor agonists carbamylcholine and norepinephrine, but not to the stimulation elicited by sodium channel agents (batrachotoxin, scorpion venom and pumiliotoxin B). Stimulation by allethrin was not additive to the stimulation elicited either by receptor agonists or sodium channel agents. A submaximal concentration of allethrin, a type I pyrethroid, did not greatly affect the dose-dependent stimulation elicited by a type II pyrethroid, deltamethrin, while a higher concentration of allethrin prevented further stimulation by type II pyrethroids. A local anesthetic, dibucaine, which inhibits sodium channel activation, inhibited phosphoinositide breakdown induced by type II, but not by type I pyrethroids, except at higher concentrations. Thus, type II pyrethroids appear to stimulate phosphoinositide breakdown in synaptoneurosomes in a manner analogous to other sodium channel agents, while type I pyrethroids elicit phosphoinositide breakdown by a different mechanism, probably not involving sodium channels. PMID- 2546658 TI - Blockade of FG 7142 kindling by anticonvulsants acting at sites distant from the benzodiazepine receptor. AB - Repeated administration of the beta-carboline FG 7142 results in sensitisation to its convulsant effects (chemical kindling); acutely FG 7142 is not convulsant, but following several treatments full seizures develop. It has been suggested that the increased sensitivity results from changes in benzodiazepine (BZ)/GABA receptor function. The present experiments studied the ability of BZ receptor ligands and anticonvulsant drugs with diverse mechanisms of action to block the expression and development of kindling to once daily injection of FG 7142 (40 mg/kg, i.p.) in mice. In fully kindled mice, the BZ receptor agonists clonazepam, ZK 93,423 and CL 218,872, and the antagonists flumazenil and ZK 93,426 prevented FG 7142 convulsions, as did 2 anticonvulsants, sodium valproate, possibly acting by influencing GABAergic transmission, and ethosuximide. A further two substances, MK 801 and 2-chloradenosine which act respectively via glutamatergic and purinergic mechanisms were also effective. When administered concomitantly with repeated FG 7142, all of these substances prevented or strongly reduced the development of kindling. Phenytoin and carbamazepine were ineffective in protecting against FG 7142 convulsions in kindled mice, and in preventing the development of kindling when administered repeatedly together with FG 7142. Since MK 801 and 2-chloradenosine prevented kindling, these results suggest that an interaction of FG 7142 with BZ receptors is not sufficient to induce kindling, which may instead result from secondary changes in sites distant from BZ/GABA receptors. PMID- 2546659 TI - Ketotifen does not inhibit asthmatic reactions induced by toluene di-isocyanate in sensitized subjects. AB - In order to determine whether treatment with ketotifen inhibits asthmatic reactions induced by toluene di-isocyanate (TDI), we studied six sensitized subjects with previously demonstrated dual or late asthmatic reaction after inhalation challenge with TDI. Ketotifen (1 mg b.i.d., orally) or placebo was administered for 7 days to the examined subjects, according to a double-blind, cross-over, placebo-controlled study design. When the subjects were treated with either ketotifen or placebo, FEV1 markedly decreased after exposure to TDI. These results suggest that the anti-asthmatic agent ketotifen is not effective in TDI induced asthma and suggest that it should not be used in the prophylaxis of asthmatic reactions induced by TDI in sensitized subjects. PMID- 2546660 TI - Periodic health examination, 1989 update: 2. Early detection of colorectal cancer and problem drinking [corrected]. PMID- 2546661 TI - Three-dimensional demonstration of the intracellular structures of mouse mesothelial cells by scanning electron microscopy. AB - Intracellular structures of mouse mesothelial cells lining the small intestine were studied by scanning electron microscopy. Specimens were prepared by the freeze-polishing method which permitted demonstration of the attenuated thin layered organization of the intracellular structures. When the surface cell membrane had been partially turned over during the specimen preparation procedure, many pinocytotic vesicles were observed attached to the cytoplasmic side of the cell membrane. We also demonstrated the external side of the basal cell membrane on which many openings of the pinocytotic vesicles were clearly shown as those recognized on the free cell surface. Two adjacent pinocytotic vesicles often appeared fused together. The multivesicular vacuole formed by the congregation of the pinocytotic vesicles were sometimes recognized on the cytoplasmic side of the surface and basal cell membrane. On the nuclear surface, nuclear pores and polysomes were visible. From our SEM findings, we classified the rough endoplasmic reticulum into three types: type I, II and III. The type I rough endoplasmic reticulum was composed of flat cisternae with fenestration, the type II of polygonal flat cisternae without fenestration, and the type III of vesicular cisternae. Polysomal formation is evident in the type III, but less in the type I and II. All of the endoplasmic reticulum appeared connected each other forming a continuous system in the cell. Two kinds of the Golgi apparatus were recognized: a stretched type with highly extended Golgi cisternae and a shrunken type with less developed Golgi cisternae. In the latter, lots of Golgi vesicles were attached on the periphery of the cisternae, and the outermost cisterna was commonly fenestrated. PMID- 2546662 TI - Are glycoconjugates and their endogenous receptors involved in the fusion of mononuclear macrophages resulting in multinucleate giant cells? Histochemical and electron microscopic determination of endogenous sugar-binding proteins (lectins) in mononuclear macrophages and multinucleate giant cells appearing in granulomatous foreign body reaction. AB - Protein-carbohydrate recognition has been found to play an important role in phagocytosis. Labelled (neo)glycoproteins were employed to comparatively analyze the histochemical pattern and ultrastructural localization of endogenous carbohydrate-binding proteins (lectins) of mononuclear macrophages and multinucleate giant cells involved in the granulomatous foreign body reaction. Sugar receptors having an affinity to simple alpha- and beta-galactoside structures, to alpha-mannose residues, to N-acetylglucosamine, to N acetylgalactosamine and to glucuronic acid, respectively, were detected in both cell types. However, alpha-fucoside- and beta-xyloside-specific receptors were present only in the mononuclear macrophages. Pronounced differences were seen with labelled, suitably modified glycoproteins, exposing different complex sugar parts with common beta-galactoside-termini. Among the population of multinucleate giant cells, a positive histochemical reaction was observed with mannose-6 phosphate-, galactose-6-phosphate- and glucuronic acid-(BSA-biotin), respectively, only in giant cells in which fusing mononuclear cells were recognizable. This transient expression indicates changes within the profile of endogenous sugar receptors in the stages from fusion to establishment of giant cells. Aside from the diffuse intracytoplasmic distribution of carbohydrate binding proteins, a prominent accumulation of various types of glycosylated ferritin, used as a marker for electron microscopic evaluation, was ultrastructurally found in membranous subcellular structures and vesicles. This study is a basis for further investigation of the potential involvement of various sugar receptors in the process of macrophage fusion, resulting in multinucleate giant cells of foreign body type, and the process of phagocytosis. PMID- 2546663 TI - Freeze-fracture study of the intercellular junctions in bronchiolo-alveolar carcinoma (Clara cell type). AB - Two bronchiolo-alveolar carcinomas of Clara cell type have been studied by thin section and freeze-fracture techniques. Thin sections of the tumour cells showed junctional complexes (tight junctions and intermediate junctions) at the luminal aspect of the lateral cell membranes and desmosomes below them. Freeze-fracture replicas showed well-developed tight junctions with strands arranged in parallel or polygonal patterns. The junctions were continuous 'zonula occludens' type, usually three or more strands deep, with focal proliferation in some areas. The granules in the tumour cells tended to be larger than those of normal human Clara cells, although they were morphologically similar. There was no evidence of secretion of granules from the tumour cells. The relationships between tumour differentiation and growth pattern, and junctional development are discussed. PMID- 2546664 TI - Ultrastructural features that relate to tumor cell behaviour in human breast carcinomas. AB - Human breast tumor biopsies were studied by transmission electron microscopy for ultrastructural characteristics that might be associated with tumor cell function and behaviour. Features related to the state of cell differentiation and tumor cell detachment were evaluated and discussed. The ultrastructure of loose cells at the tumor periphery is compared with that of coherent cells in interior regions. Structural information has been analysed by taking into account available biochemical data for interpretation of cell function. The value of the parameters chosen for study is discussed. This study shows that each tumor needs to be evaluated independently for its specific ultrastructural features. Electron microscopic data becomes more relevant when interpreted in conjunction with histological diagnosis and clinical findings. PMID- 2546665 TI - Stereological and functional investigations on isolated adrenocortical cells. I. Adrenocortical cells of normal adult rats. AB - The function and morphology of isolated rat adrenocortical cells were investigated. Capsular (zona glomerulosa) cells secreted aldosterone, progesterone and small amounts of corticosterone; inner (zonae fasciculata and reticularis) cells produced corticosterone and progesterone. ACTH (10(-8) M) exposure caused a striking rise in aldosterone and corticosterone secretion by capsular cells, and in corticosterone release by inner cell preparations; simultaneously, progesterone secretion was significantly reduced. Capsular and inner cells possessed an average volume of 500 and 1250 micron 3, and ultrastructural qualitative and quantitative (stereological) features closely resembling those of zona glomerulosa and zona fasciculata-reticularis cells in situ, respectively. ACTH exposure caused a notable lipid-droplet depletion in both types of isolated cells. PMID- 2546666 TI - Chromosome changes in a brain metastasis of a large cell lung cancer. AB - Direct chromosome preparations were made on a brain metastasis of a large cell lung cancer from a 46-year-old female patient. In the 169 cells examined, the modal chromosomes number was 73, and the majority (53.2%) of mitoses were in the near-triploid region. The tumor cells showed considerable heterogeneity in chromosome number and content. Chromosomes 3, 4, and 5 were the most frequently involved in aberrations. The mean number of unidentified marker chromosomes was five per mitosis. PMID- 2546667 TI - The relation between inhibition of cell growth and of dihydroorotic acid dehydrogenase by brequinar sodium. AB - The growth inhibitory effects of Brequinar Sodium (DUP-785; NSC 368390) in 7 different cell lines were related to growth rates and to the inhibition of dihydroorotic acid dehydrogenase (DHO-DH) activity. IC50 values were between 0.2 and 5.8 microM; the fastest growing cell line was least sensitive. Despite a large variation in sensitivity, basal activity of DHO-DH showed little variation (only 2-fold) between the different cell lines. Residual activity of DHO-DH in the presence of Brequinar Sodium varied 30-fold. Drug sensitivity correlated with this residual DHO-DH activity; DHO-DH activity was only slightly inhibited by Brequinar Sodium in the most resistant lines, and almost completely in the most sensitive. PMID- 2546668 TI - G.L.C.-M.S. of N-(1-deoxyalditol-1-yl)octadecylamine derivatives in the analysis of methanolysates of neoglycolipids obtained by reductive amination. AB - Hydrophobic conjugates of a series of aldoses have been prepared by reductive amination with octadecylamine and sodium cyanoborohydride, as model compounds for the analysis of reductively aminated oligosaccharides derived from capsular polysaccharides of Streptococcus pneumoniae. In the context of the methanolysis procedure for sugar analysis, g.l.c. and g.l.c.-m.s. (e.i.-mode) studies were carried out on the N-(1-deoxyalditol-1-yl)octadecylamine derivatives obtained after treatment with methanolic HCl, and subsequent N-acetylation and trimethylsilylation. PMID- 2546669 TI - A dorsomedial subdivision within the nucleus intercollicularis identified in the Japanese quail (Coturnix coturnix japonica) by means of alpha 2-adrenergic receptor autoradiography and estrogen receptor immunohistochemistry. AB - The nucleus intercollicularis is an important site in the control of vocalization in birds. In oscines, a subregion of the nucleus intercollicularis called the dorso-medial intercollicular nucleus appears to play a key role in this process because it receives the majority of the projections from the nucleus robustus archistriatalis and sends most of the projections to the motor nucleus of the hypoglossal nerve. In this paper, we present neurochemical studies of the nucleus intercollicularis in the Japanese quail which suggest the presence of heterogeneity within this structure. One rostral band contains high densities of cholinergic muscarinic receptors identified by quantitative autoradiography using tritiated N-methylscopolamine as the ligand. A caudal dorso-medial region is specifically labeled by estrogen receptors identified using immunocytochemistry and by alpha 2-adrenergic receptors which were quantified by autoradiography using tritiated para-amino-clonidine. This latter subregion is possibly equivalent to the dorso-medial intercollicular nucleus of oscines. Additional track-tracing studies should be performed to confirm this homology. The coexistence of estrogen and alpha 2-adrenergic receptors within the same structure suggests important functional connections between steroid action and catecholaminergic systems in the brain. PMID- 2546670 TI - Localisation of islet amyloid peptide in lipofuscin bodies and secretory granules of human B-cells and in islets of type-2 diabetic subjects. AB - Islet amyloid peptide (or diabetes-associated peptide), the major component of pancreatic islet amyloid found in type-2 diabetes, has been identified by electron-microscopic immunocytochemistry in pancreatic B-cells from five non diabetic human subjects, and in islets from five type-2 diabetic patients. The greatest density of immunoreactivity for islet amyloid peptide was found in electron-dense regions of some lysosomal or lipofuscin bodies. The peptide was also localised by quantification of immunogold in the secretory granules of B cells, and was present in cytoplasmic lamellar bodies. Acid phosphatase activity was also demonstrated in these organelles. Immunoreactivity for insulin was found in some lysosomes. These results suggest that islet amyloid peptide is a constituent of normal pancreatic B-cells, and accumulates in lipofuscin bodies where it is presumably partially degraded. In islets from type-2 diabetic subjects, amyloid fibrils and lipofuscin bodies in B-cells showed immunoreactivity for the amyloid peptide. Abnormal processing of the peptide within B-cells could lead to the formation of islet amyloid in type-2 diabetes. PMID- 2546671 TI - Gin-mediated recombination of catenated and knotted DNA substrates: implications for the mechanism of interaction between cis-acting sites. AB - The Gin DNA-inversion system of bacteriophage Mu normally requires a substrate containing two inverted recombination sites (gix) and an enhancer sequence on the same supercoiled DNA molecule. The reaction mechanism was investigated by separating these sites on catenated rings. Catenanes with the gix sites on one circle and the enhancer on the other recombined efficiently. Thus, the enhancer was fully functional even though it was located in trans to the gix sites. Multiple links between the rings are required for recombination. Multiply linked catenanes with gix sites on separate circles, one of which contained the enhancer, were also efficient substrates. Knotted constructs carrying directly repeated gix sites were recombined. Catenated and knotted substrates must also be supercoiled. These experiments eliminate simple tracking or looping models as explanations for why the enhancer and gix sites must be in cis with standard substrates. Rather, the Gin synaptic complex requires the three sites to be mutually intertwined in a right-handed fashion with a unique polarity of the gix sites. This geometry is achieved by branching of the DNA substrate and requires the energy and structure of supercoiling, catenation, or knotting. PMID- 2546672 TI - Purification and characterization of CAF-I, a human cell factor required for chromatin assembly during DNA replication in vitro. AB - The purification and characterization of a replication-dependent chromatin assembly factor (CAF-I) from the nuclei of human cells is described. CAF-I is a multisubunit protein that, when added to a crude cytosol replication extract, promotes chromatin assembly on replicating SV40 DNA. Chromatin assembly by CAF-I requires and is coupled with DNA replication. The minichromosomes assembled de novo by CAF-I consist of correctly spaced nucleosomes containing the four core histones H2A, H2B, H3, and H4, which are supplied in a soluble form by the cytosol replication extract. Thus, by several criteria, the CAF-I-dependent chromatin assembly reaction described herein reflects the process of chromatin formation during DNA replication in vivo. PMID- 2546673 TI - Structure of the termini of DNA intermediates in the integration of retroviral DNA: dependence on IN function and terminal DNA sequence. AB - Linear retroviral DNA, the major precursor to the integrated provirus of the murine leukemia viruses, contains a mixture of two structures at its ends: some termini are full-length and blunt, and some have recessed 3' strands. A temporal study of the end structures showed that the proportion of the DNA with recessed ends increases during the course of infection, and suggests that the blunt ends are precursors to the recessed ends. We have examined the DNA structures of the ends of retroviral mutants defective in the integration (IN) function. The results show that the formation of the recessed ends requires the presence of IN. Finally, we have analyzed the structures at the ends of mutant genomes with alterations in the terminal DNA sequence. The exact position of the recessed 3' end can be recessed one, two, or four nucleotides relative to the 5' end. In all cases the position of the recessed 3' end correlates perfectly with, and thus presumably determines, the site of joining to the target DNA. PMID- 2546674 TI - Viral latency and transformation: the strategy of Epstein-Barr virus. PMID- 2546675 TI - A new twist to the topoisomerase I problem. PMID- 2546676 TI - G protein linked signal transduction pathways in development: dictyostelium as an experimental system. PMID- 2546677 TI - Positive autoregulation of the myogenic determination gene MyoD1. AB - Transfection of cDNA expression vectors encoding either MyoD1 or myogenin into 10T1/2 cells converts them to myogenic cells. We show that transfection of 10T1/2 cells with the MyoD1 cDNA activates expression of endogenous MyoD1 mRNA, indicating that MyoD1 is subject to positive autoregulation. This activation of endogenous MyoD1 mRNA was also observed in Swiss 3T6 cells, but not in several other fibroblast or adipoblast cell lines transfected with the MyoD1 cDNA. In addition, transfection of the MyoD1 cDNA leads to activation of myogenin expression, and transfection of the myogenin cDNA leads to activation of MyoD1 expression. Thus, MyoD1 and myogenin appear to function in a positive autoregulatory loop that could either: account for or contribute to the stability of myogenic commitment; or amplify the level of expression of both MyoD1 and myogenin above a critical threshold that is required for activation of the myogenic program. PMID- 2546678 TI - The cellular 107K protein that binds to adenovirus E1A also associates with the large T antigens of SV40 and JC virus. AB - The association between the retinoblastoma protein (p105-RB) and either the large T antigen of SV40 or the E1A proteins of adenovirus is thought to be an important step in transformation by these viral oncogenes. E1A and large T antigen share a small region of amino acid homology that is necessary for high affinity binding with p105-RB. Mutations of this homology region were shown to reduce drastically the frequency of transformation mediated by the E1A or large T oncogenes. Previously, this small region in E1A was shown to be sufficient for interaction with a second cellular protein of 107,000 daltons (107K). Here we show that in human cells, the large T antigens of SV40 or JC virus also form complexes with 107K. Demonstration of complexes between 107K and the large T antigens of SV40 and JC virus suggests that these associations may represent another component of a common mechanism for transformation between adenoviruses and polyoma viruses. PMID- 2546679 TI - HIV-1 TAT "activates" presynthesized RNA in the nucleus. AB - Replication of the human immunodeficiency virus (HIV-1) depends upon the viral TAT protein. TAT stimulates gene expression via a target response sequence (TAR) located within the HIV-1 LTR. As TAR is located in the transcribed region it could act as a signal in either the DNA, the RNA, or both. To test whether TAT acts on transcription and/or posttranscriptionally, we produced TAT in yeast and monitored its activity after microinjection into the nucleus or cytoplasm of Xenopus oocytes. The TAT protein stimulated TAR-dependent expression, but this activation was not inhibited by transcriptional inhibitors. Furthermore, TAR containing RNA, produced in vitro, was "activated" by TAT after coinjection into oocytes. This activation only occurred, however, when the RNA was injected into the nucleus and not into the cytoplasm. Our data indicate, therefore, that in the Xenopus system TAT acts on presynthesized RNA and that the nucleus is involved in this action. PMID- 2546680 TI - Specific and saturable binding of pp60v-src to plasma membranes: evidence for a myristyl-src receptor. AB - The molecular basis for membrane association of pp60v-src, the transforming protein of Rous sarcoma virus, was investigated in a cell-free system. Newly synthesized pp60v-src polypeptide, produced by in vitro translation of src mRNA, rapidly bound to plasma membranes. Binding was saturable and dependent on the presence of myristate at the amino terminus of pp60v-src. Prior treatment of membranes with heat or trypsin greatly decreased subsequent binding of pp60v-src. Membrane binding of pp60v-src was competed by a myristylated peptide containing the first 11 amino acids of the mature src sequence, but not by non-myristylated src peptide or other myristylated peptides. The specificity, saturability, and competitive nature of pp60v-src binding provide evidence for the existence of a src receptor in the plasma membrane. PMID- 2546681 TI - Efficient Mu transposition requires interaction of transposase with a DNA sequence at the Mu operator: implications for regulation. AB - Phage Mu transposition is initiated by the Mu DNA strand-transfer reaction, which generates a branched DNA structure that acts as a transposition intermediate. A critical step in this reaction is formation of a special synaptic DNA-protein complex called a plectosome. We find that formation of this complex involves, in addition to a pair of Mu end sequences, a third cis-acting sequence element, the internal activation sequence (IAS). The IAS is specifically recognized by the N terminal domain of Mu transposase (MuA protein). Neither the N-terminal domain of MuA protein nor the IAS is required for later reaction steps. The IAS overlaps with the sequences to which Mu repressor protein binds in the Mu operator region; the Mu repressor directly inhibits the Mu DNA strand-transfer reaction by interfering with the interaction between MuA protein and the IAS, providing an additional mode of regulation by the repressor. PMID- 2546682 TI - A hyper-recombination mutation in S. cerevisiae identifies a novel eukaryotic topoisomerase. AB - A hyper-recombination mutation was isolated that causes an increase in recombination between short repeated delta sequences surrounding the SUP4-omicron gene in S. cerevisiae. The wild-type copy of this gene was cloned by complementation of one of its pleiotropic phenotypes, slow growth. DNA sequence of the clone revealed a 656 amino acid open reading frame capable of encoding a protein homologous to the bacterial type I topoisomerase. No homology was detected with previously identified eukaryotic topoisomerases. Construction of double mutants with either of the two known yeast topoisomerase genes revealed synergistic effects on growth suggesting overlapping functions. Expression of bacterial topoisomerase I in yeast can fully complement the slow growth defect of a null mutation. We have named this locus TOP3 and suggest that it defines a novel eukaryotic topoisomerase gene. PMID- 2546683 TI - A mitochondrial frameshift suppressor maps in the tRNASer-var1 region of the mitochondrial genome of the yeast S. cerevisiae. AB - A polypeptide chain-terminating mutation (M5631) previously has been shown to be a +1T insertion in the yeast mitochondrial gene oxi1, coding for subunit II of the cytochrome c oxidase. A spontaneously arisen frameshift suppressor (mfs-1) that is mitochondrially inherited suppresses this mutation to a considerable extent. The suppressor mutation was mapped by genetic and molecular analyses in the mitochondrial tRNASer-var1 region of the mitochondrial genome of the yeast S. cerevisiae. Genetic analyses show that the suppressor mfs-1 does not suppress other known mitochondrial frameshift mutations, or missense and nonsense mutations. PMID- 2546684 TI - Functional relationship among TATA sequences, gene induction and transcription initiation in the beta-galactosidase, LAC4, gene from Kluyveromyces lactis. AB - In the 5' non-coding region of the beta-galactosidase, LAC4, gene of Kluyveromyces lactis, three TATA-like sequences are present at -230, -170 and 142 from the ATG translation start site. By means of deletion mutations in the TATA region, at least two of these TATA sequences, those at -230 and -142, were shown to be required for normal gene expression. Evidence is presented for a functional hierarchy and cooperation between these TATA sequences. The deletion or a change in the position of the TATA sequences affects both beta-galactosidase induction and the location of RNA initiation sites. The TATA sequence at -230 alone is sufficient for correct gene induction when it is moved to a position 41 bp from the major RNA initiation sites located around -110; the -142 TATA alone contributes only partly to gene induction. We suggest a functional distinction between these two related regulatory sequences. This functional distinction might be established by sequence differences and/or targets of unlike specific DNA binding protein(s). A conformational analysis of the LAC4 promoter showed that under torsional stress the functional elements UAS, TATA boxes RNA initiation sites and ATG can be detected as P1-sensitive sites. Possible functions of DNA structural alterations on gene expression are discussed. PMID- 2546685 TI - Restricted distribution of the Tad transposon in strains of Neurospora. AB - A simple colony blot procedure was used to screen 336 Neurospora strains for the presence of the transposable element Tad. These strains included the standard laboratory wild types, all of the available Neurospora isolates collected from the Ivory Coast, and all wild-collected Neurospora crassa isolates available from the Fungal Genetics Stock Center. Tad was found only in the strain of origin from Adiopodume, Ivory Coast, where it is present in multiple copies. Three other strains of African origin were found to have single copy sequences that are related to Tad. PMID- 2546686 TI - A controversy concerning the structure of the mitochondrial genome of a yeast petite mutant: resolution by sequence analysis. AB - Sequence analysis was used to define the repeat unit that constitutes the mitochondrial genome of a petite (rho-) mutant of the yeast Saccharomyces cerevisiae. This mutant has retained and amplified in tandem a 2,547 bp segment encompassing the second exon of the oxi3 gene excised from wild-type mtDNA between two direct repeats of 11 nucleotides. The identity of the mtDNA segment retained in this petite has recently been questioned (van der Veen et al., 1988). The results presented here confirm the identity of this mtDNA segment to be that determined previously by restriction mapping (Carignani et al., 1983). PMID- 2546687 TI - Anticoagulant activity of immobilized thrombomodulin. AB - Bovine lung thrombomodulin was partially purified, and immobilized on agarose gel (Sepharose 4B). Immobilized thrombomodulin inhibited the procoagulant activity of thrombin, and enhanced the thrombin-catalyzed protein C activation. The plasma recalcification time test showed that immobilized thrombomodulin prolonged plasma clotting time. It is suggested that the immobilization of thrombomodulin will provide an antithrombogenic biomaterial able to convert thrombin from a procoagulant to an anticoagulant enzyme. PMID- 2546688 TI - Severe renal toxicity due to intermediate-dose methotrexate. AB - Methotrexate (MTX) is a drug widely used in the treatment of patients with malignant disease. Its well-known side effects include myelosuppression, mucositis and renal damage. These problems are primarily dose-related, tending to occur more frequently when high doses (greater than 1 g/m2) are given. We present four cases in whom severe renal and mucosal toxicity occurred with intermediate doses (200 mg/m2) of MTX despite folinic acid rescue. Possible reasons for this occurrence are discussed and means of avoiding such toxicity are suggested. Three of four patients developed severe loin pain within a few hours of injection; the significance of this symptom in relation to subsequent renal toxicity has implications for early recognition of the problem. PMID- 2546689 TI - Stimulated production of ATP by H2O2 disproportionation in extracts from normal and xeroderma pigmentosum skins, and from normal, xeroderma pigmentosum, ataxia telangiectasia and simian virus 40 transformed cell lines. AB - It has been previously shown that xeroderma pigmentosum (XP) skin biopsies and their established cell lines exhibit a decrease in catalase activity and enhanced formation of photo-produced H2O2. Several in vivo and in vitro thermodynamic results suggest that the energy of H2O2 disproportionation produced by catalase could be sufficient to synthesize ATP with or without the help of intact mitochondria. In this paper, we first studied the properties of H2O2-stimulated ATP production in extracts of normal and pathological XP skin biopsies and cell lines. In acellular extracts of normal skin biopsies and/or cell lines, ATP production can be increased 2- to 3-fold, but only with a narrow range of H2O2 concentration. In contrast, in extracts of pathological skins or cells, ATP production was only observed when using 10- to 1000-fold less H2O2 concentration as defined for normal extracts. Similar results were noted with two cell lines derived from patients afflicted with ataxia telangiectasia (AT), and with simian virus 40 (SV40) transformed lines of normal, XP and AT cells, Although we have no proof that such a process may exist in vivo, we would like to suggest that both H2O2-stimulated ATP production and catalase activity are good indicators of the degree of normality or abnormality of skin biopsies and/or cell lines. PMID- 2546690 TI - Cycles of repeated augmentation pressure in rapid production of pancreatic and cholangiocellular carcinomas in hamsters initiated with N-nitrosobis(2 oxopropyl)amine. AB - We previously reported a rapid production model for pancreatic carcinoma development in Syrian hamsters incorporating the principle of selection by resistance to cytotoxicity. In the present experiment, the efficacy of repeated augmentation pressure with regard to generation of pancreatic lesions in hamsters initiated with N-nitrosobis(2-oxopropyl)amine (BOP) was investigated. Forty-eight female Syrian golden hamsters were divided into four groups according to the frequency of augmentation pressure. Group 1 received 70 mg/kg body weight of BOP and three injections of 20 mg/kg BOP. Groups 2-4 received 70 mg/kg BOP followed by one, two or three cycles of augmentation pressure consisting of dl-ethionine on sugar and salt diet, l-methionine and 20 mg/kg BOP. Hamsters were killed 10 weeks after the beginning of the experiment and the resultant incidences of pancreatic carcinomas from groups 1-4 were 0, 30, 50 and 46.2% respectively, the numbers of pancreatic carcinomas increasing with the frequency of augmentation pressure. A 46.2% yield of cholangiocarcinomas was also observed in group 4. The model should be useful for investigation of potential modulating factors since large numbers of lesions can be induced within a total experimental period of only 10 weeks. PMID- 2546691 TI - Atrial natriuretic peptide in heart and specific binding in organs from fetal and newborn rats. AB - To assess the possibility that atrial natriuretic peptide plays a role in salt and water balance during early mammalian development, we examined hearts from fetal and neonatal rates for the presence of this peptide and presumed target tissues for their ability to bind the hormone. Immunohistochemistry was used to localize and radioimmunoassay to quantify this peptide in heart. Immunoreactive atrial natriuretic peptide was visualized in the fetal heart on day 17.5 post conception. It was distributed throughout the atrial appendages and free wall and, in ventricle, in the trabeculae carnae and chordae tendineae. The concentrations of immunoreactive atrial natriuretic peptide in atria of rats on day 19.5 post-conception were one-tenth of those in the adult. Levels of this peptide in fetal ventricle were low and virtually absent from the adult tissue. Specific binding of radiolabelled atrial natriuretic peptide measured by whole organ counting occurred in several organs from 19.5-day fetal and neonatal rats. A number of these tissues, including the kidney, ileum, adrenal, lung and liver, are targets for and/or bind the peptide in adult rats. Specific binding in these tissues was localized using autoradiography at anatomical sites similar to those in adult organs. Specific binding was also seen in fetal but not neonatal skin. In the kidney, binding was associated with immature as well as mature glomeruli. These findings support the proposition that atrial natriuretic peptide may function in the perinatal rat as it does in the adult and, in addition, may play a unique role during fetal life. PMID- 2546692 TI - Ionic requirements of the endothelin response in aorta and portal vein. AB - The vasoconstrictor responses of isolated rat portal vein and aorta to synthetically prepared endothelin are investigated. Both preparations respond to 10(-9) M levels of the peptide although the aortic response is more sustained than that of the portal vein. Endothelin-evoked contractions, unlike those evoked by scorpion alpha-toxins (which are homologous to endothelin) or by veratridine, are insensitive to tetrodotoxin or to the removal of sodium ions from the tissue bathing medium. Contractile responses to endothelin may still be observed in high potassium depolarizing medium and are not dependent on the presence of extracellular chloride; however, the responses are dependent on the presence of extracellular calcium and are blocked by nitrendipine, nifedipine, or nickel. Endothelin-evoked uptake of 45Ca into aortic tissue is also independent of extracellular sodium or potassium and is blocked by nifedipine. These data strongly suggest that endothelin acts at a site closely coupled to the calcium channel and that depolarization by sodium influx through voltage-dependent channels is not involved in endothelin-induced vasoconstriction. PMID- 2546693 TI - Vasoactive intestinal peptide receptor in failing human ventricular myocardium exhibits increased affinity and decreased density. AB - We investigated vasoactive intestinal peptide (VIP)-receptor pharmacology in failing and nonfailing human ventricular myocardium by examining [125I]VIP binding in membrane fractions of left ventricle and inotropic effects of VIP in isolated right ventricular trabeculae mounted in tissue baths. [125I]VIP binding demonstrated upwardly concave, curvilinear Scatchard plots consistent with two classes of binding sites. Only the high-affinity (dissociation constant [Kd] 400 800 pM) site could be regulated by guanine nucleotides. Compared with nonfailing heart, membranes derived from failing heart exhibited a twofold reduction in the Kd of the high-affinity VIP binding site, whereas the receptor density (Bmax) was decreased by 62%. In concordance with this decreased receptor density and increased affinity, the maximal contractile response of right ventricular trabeculae from failing right ventricles was decreased by 61%, and the dose response curve to VIP was left-shifted approximately threefold. We conclude that the VIP receptor in failing human ventricular myocardium exhibits novel regulatory behavior consisting of increased receptor affinity and decreased receptor density. PMID- 2546694 TI - Developmental changes in the electrophysiological properties and the beta adrenergic receptor-effector complex in atrial fibers of the canine coronary sinus. AB - beta-Adrenergic stimulation induced delayed afterdepolarizations and triggered activity in atrial fibers of adult but not young canine coronary sinus. However, sensitivity to beta-adrenergic agonists with respect to maximum diastolic potential was identical at both ages, and delayed afterdepolarizations and triggered activity did occur in response to ouabain. Age-dependent lengthening of the action potential duration and plateau also were seen and were greater in the adult than the young. beta-Adrenergic receptor density and affinity and the stimulatory guanine nucleotide regulatory protein (Gs) were similar in adult and young tissues. In contrast, the inhibitory guanine nucleotide regulatory protein (Gi) was 2.5-fold greater in adult (15 fmol/mg) than in young (6.0 fmol/mg) tissues. Basal- and forskolin-stimulated adenylate cyclase activities were greater in adult than young coronary sinus although the increment in isoproterenol-stimulated adenylate cyclase activity in young tissue was greater when compared either with basal levels or expressed as a percentage of maximal catalytic activity. Both the traditional effector pathway of beta-adrenergic action, involving the stimulation of adenylate cyclase activity, and developmental changes in the action potential plateau may contribute to the developmental changes in delayed afterdepolarizations and triggered activity. PMID- 2546695 TI - Relaxation of rabbit ventricular muscle by Na-Ca exchange and sarcoplasmic reticulum calcium pump. Ryanodine and voltage sensitivity. AB - We studied relaxation during rapid rewarming of rabbit ventricular muscles that had been activated by rapid cooling. Rewarming from 1 degree to 30 degrees C (in less than 0.5 second) activates mechanisms that contribute to the reduction of intracellular calcium concentration and thus relaxation (e.g., sarcoplasmic reticulum [SR] calcium pump and sarcolemmal Na-Ca exchange and calcium pump). Rapid rewarming in normal Tyrode's solution induces relaxation with a half-time (t1/2) of 217 +/- 14 msec (mean +/- SEM). During cold exposure, changing the superfusate to a sodium-free, calcium-free medium with 2 mM CoCl2 (to eliminate Na-Ca exchange) slightly slows relaxation upon rewarming in the same medium (t1/2 = 279 +/- 44 msec). Addition of 10 mM caffeine (which prevents SR calcium sequestration) to normal Tyrode's solution during cold superfusion slows relaxation somewhat more (t1/2 = 376 +/- 31 msec) than sodium-free, calcium-free solution. However, if both interventions are combined (sodium-free + caffeine) during the cold exposure and rewarming, the relaxation is greatly slowed (t1/2 = 2,580 +/- 810 msec). These results suggest that either the SR calcium pump or, to a lesser extent, sarcolemmal Na-Ca exchange can produce rapid relaxation, but if both systems are blocked, relaxation is very slow. If muscles are equilibrated with 500 nM ryanodine before cooling, relaxation upon rewarming is not greatly slowed (t1/2 = 266 +/- 37 msec) even if sodium-free, calcium-free solution is applied during the cold and rewarming phases (t1/2 = 305 +/- 66 msec). This result suggests that ryanodine does not prevent the SR from accumulating calcium to induce relaxation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546696 TI - The effect of acute alterations in left ventricular afterload and beta-adrenergic tone on indices of early diastolic filling rate. AB - The effects of an acute increase in left ventricular systolic pressure and the effects of an intravenous isoproterenol infusion on myocardial (segment) lengthening rate and chamber (minor axis dimension) filling rate were examined in 12 anesthetized dogs. Measurements of left ventricular systolic pressure (by micromanometer) and of segment length and chamber dimension transients (by ultrasonic crystals) were made in variably afterloaded beats (three-beat descending aortic cross-clamp) before and during an isoproterenol infusion that raised (+)dP/dt by 40%. During the baseline state, we found an inverse relation between the peak rate of increase in minor axis dimension [(+)dD/dt] and systolic pressure over a wide range of systolic pressures (110-160 mm Hg) and end-systolic dimensions (25-40 mm); peak (+)dD/dt and end-systolic dimension were also inversely related. During isoproterenol infusion, end-systolic dimension fell from 29.7 +/- 3.1 to 28.0 +/- 3.1 mm and (+)dD/dt increased from 79.6 +/- 8.0 to 90.1 +/- 8.7 mm/sec; however, the slope and y intercept of the relation between (+)dD/dt and end-systolic dimension were unchanged. Peak (+)dD/dt at a common end systolic dimension of 31 mm was nearly equal during baseline and isoproterenol states (64.2 +/- 6.3 vs. 65.1 +/- 6.6 mm/sec). Similar results were found using segment length transients. We interpret these data to indicate that (+)dD/dt is strongly influenced by changes in systolic pressure and dimension and that isoproterenol-induced changes in (+)dD/dt are mediated, at least in part, through changes in systolic pressure and dimension. PMID- 2546697 TI - Amiodarone: biochemical evidence for binding to a receptor for class I drugs associated with the rat cardiac sodium channel. AB - Amiodarone has multiple pharmacological effects in heart. Electrophysiological data suggest that among its other effects, amiodarone is a sodium channel blocker. Using a radioligand assay, we determined whether amiodarone interacted with a previously described receptor for type I agents associated with the cardiac sodium channel. The radioligand was [3H]batrachotoxinin A 20 alpha benzoate ([ 3H]BTXB), a toxin that binds to the activated state of the sodium channel. We have previously shown that class I antiarrhythmic drugs inhibit [3H]BTXB binding. The purpose of this study was to assess whether amiodarone and other class III agents interact with this receptor. Amiodarone inhibited [3H]BTXB binding in a dose-dependent fashion, with an estimated IC50 value of 3.6 microM. This IC50 value is similar to reported clinically effective serum concentrations of amiodarone. In contrast to amiodarone, the IC50 values for other class III drugs (bretylium, sotalol, bethanidine, N-acetylprocainamide) were much higher than their therapeutic concentrations and bore no relation to them. Scatchard analysis of [3H]BTXB binding showed that amiodarone reduced the maximal binding for [3H]BTXB; this finding indicates irreversible inhibition or (more likely) allosteric inhibition by amiodarone. The latter agrees with electrophysiological data suggesting that amiodarone binds to inactivated sodium channels. Sodium channel blockade by amiodarone may contribute to its overall electrophysiological effect. PMID- 2546699 TI - Inhibition of neutrophil and natural killer cell function by human seminal fluid acid phosphatase. AB - The major acid phosphatase of human seminal fluid was purified to homogeneity by chromatography on Sephadex G-150, and DEAE-Sephadex, and by isoelectric focusing (pI, 4.3). This purified preparation of seminal fluid acid phosphatase blocked superoxide anion production by neutrophils stimulated with fMet-Leu-Phe (fMLP) by 50%. The phosphatase also hydrolysed myo-inositol 1,4,5-trisphosphate (IP3) in vitro, an intracellular second messenger which releases Ca2+ from intracellular pools, at nearly one-third the rate at which the nonphysiologic substrate 4 methylumbelliferylphosphate (MUP) was cleaved. In contrast, two phosphoinositide lipids, namely phosphatidylinositol 4,5-bisphosphate and phosphatidylinositol 4 monophosphate, were poor phosphatase substrates. Following fMLP stimulation of [3H]inositol-labeled neutrophils, the quantity of IP3 produced by phosphatase treated cells was reduced by 69%. These results indicate that the human seminal fluid acid phosphatase may compromise the neutrophil's microbicidal response to the organism by hydrolyzing a second messenger (IP3) which is directly involved in the regulation of intracellular Ca2+ concentrations. The seminal fluid phosphatase also inhibited by 85% the ability of murine natural killer (NK) cells to inactivate target cells. PMID- 2546698 TI - Impaired chronotropic response to exercise in patients with congestive heart failure. Role of postsynaptic beta-adrenergic desensitization. AB - The mechanism responsible for the attenuated heart rate (HR) response to exercise in patients with congestive heart failure (CHF) was investigated in 46 normal subjects and 59 patients with CHF stratified by peak exercise oxygen consumption (VO2). The peak exercise HR and the increment in HR from rest to peak exercise were decreased in CHF patients, and both correlated strongly with peak VO2 (r = 0.810, p less than 0.0001; r = 0.863, p less than 0.0001, respectively). Peak exercise norepinephrine level (NE) and the increment in NE from rest to peak exercise were not attenuated in CHF patients. Resting NE was elevated in CHF patients and correlated inversely with peak VO2 (r = -0.595, p less than 0.001). However, no significant correlation occurred between peak VO2 and either peak exercise NE or the exercise increment in NE. The ratio of the exercise increments in HR and NE, and indirect index of sinoatrial node sympathetic responsiveness, was markedly reduced in CHF patients and was inversely related to the severity of exercise impairment. Likewise, the HR response to a graded isoproterenol infusion was markedly reduced in CHF patients. Age-matching of normal subjects and CHF patients did not affect the foregoing observations. Infusion of CHF patients with the phosphodiesterase inhibitor milrinone caused a significant increase in the ratio of the exercise increments in HR and NE. These data strongly suggest that the attenuated HR response to exercise in CHF patients is due, at least in part, to postsynaptic desensitization of the beta-adrenergic receptor pathway. PMID- 2546700 TI - Effects of N-hydroxyethyl-1-deoxynojirimycin (BAY m 1099) on the activity of neutral- and acid alpha-glucosidases in human fibroblasts and HepG2 cells. AB - The effect of the glucose analogue N-hydroxyethyl-1-deoxynojirimycin (BAY m 1099) on the activity of alpha-glucosidases was studied in human fibroblasts and HepG2 cells. BAY m 1099 inhibits neutral and acid alpha-glucosidase activities of both cell types in a dosage-dependent and reversible manner. Inhibition of endoplasmic reticulum glucosidases I and/or II is suggested by delayed processing of lysosomal (acid) alpha-glucosidase. Competitive inhibition of mature acid alpha glucosidase leads to lysosomal accumulation of glycogen as in glycogenosis type II. There seems to be little risk, however, of inducing this storage disorder when using the drug in a dose of 50 mg per os for treatment of type II diabetes. In high doses, the drug may prove useful for studying the pathogenesis of glycogenosis type II in vitro or in animal models. PMID- 2546702 TI - Mechanisms other than polyclonal B cell activation possibly involved in Epstein Barr virus-induced autoimmunity. AB - In order to verify whether Epstein-Barr virus (EBV)-induced polyclonal B cell activation is the major cause of autoimmunity during infectious mononucleosis (IM), we have investigated, by immunoblotting, the fine specificity of anti smooth muscle autoantibodies (autoAbs) in the sera of IM patients. Furthermore, we have isolated a number of in vivo infected EBV-positive cell lines from a patient with IM and compared the reactivity of the secreted immunoglobulins (Igs) with that of serum autoAbs. The reactivity of anti-smooth muscle autoAbs was found to be closely restricted to three proteins of approximate molecular weights 54, 52 and 48 kD. Furthermore, none of 48 EBV-positive B cell lines shared any reactivity with serum autoantibodies. Taken together, these results suggest that EBV-induced autoimmunity is not a consequence of a random activation of B cells, but a specific phenomenon, requiring mechanisms other than polyclonal B cell activation. PMID- 2546701 TI - Serum monoclonal antibodies derived from patients with multiple myeloma react with mycobacterial phosphoinositides and nuclear antigens. AB - The sera of 46 patients with multiple myeloma were examined for the presence of anti-tuberculosis (TB) glycolipids antibodies. In positive sera samples, anti polynucleotides, anti-histones, anti-cardiolipin, anti-Sm and anti-RNP activities were sought. Antibodies against three different mycobacterial glycolipids were detected in 10 of the 46 sera. Three (P429, P557, P5) showed high titres of antibodies against all three different TB glycolipids. Of the 10 reactive samples, two antibodies were purified (P429, P557). P557 reacted with various polynucleotides and other antigens tested (Sm, RNP, cardiolipin, histones). One immunoglobulin (P207) which showed high activity against Sm and RNP had no activity against TB glycolipids and was employed as a control. The cross reactivity between mycobacterial glycolipids and the nuclear antigens was further established by bi-directional competition assays with P429 and P557. Our study shows a high incidence (22%) of anti-TB glycolipids antibodies in sera of patients with monoclonal gammopathies, some of which show anti-DNA and other anti nuclear antigens activities. This is additional evidence for the mycobacterial nuclear antigen cross-reactivity which may suggest a possible role of infection (e.g., tuberculosis) in autoimmune diseases. PMID- 2546704 TI - Histiocytic syndromes. AB - The histiocytic syndromes are currently divided into two major categories: Langerhans cell histiocytosis and non-Langerhans cell histiocytosis. The disease entities recognized under these categories are discussed. The discussion includes clinical features, histopathology, and treatment. PMID- 2546705 TI - Cutaneous signs of AIDS. AB - In this article, selected aspects of human immunodeficiency virus (HIV) infection are reviewed, particularly as it relates to cutaneous signs and symptoms. Included are discussions of the classification, epidemiology, and clinical manifestations of this complex infectious process. PMID- 2546703 TI - HLA class II mRNA accumulation by activated human T cells following growth in conditioned medium. AB - The expression of HLA Class II beta genes by seven CD4- or CD8-positive T cell clones grown in conditioned medium was examined on the mRNA and cell surface levels. Monoclonal antibodies directed against Class II glycoproteins demonstrated the presence of these molecules on the cell surface. In addition, Northern blot analysis demonstrated that cDNA probes for the DR, DP, and DQ beta genes hybridized to mRNA isolated from each clone. Because we were interested in differences in expression of relative levels of DR, DQ and DP, mRNA isolated from the clones was quantified by means of densitometry. Our results demonstrated that different ratios of DR, DQ and DP beta chain messages were expressed by the clones. Subsequent studies established the kinetics of the accumulation of DR beta mRNA messages by T cells during culture and showed that it could be regulated by factors contained within conditioned medium. Our results confirm that the accumulation of Class II mRNA following growth of T cells in conditioned medium appears to be a general feature. PMID- 2546706 TI - [31P-NMR spectroscopy of mitochondrial myopathies: the relation between abnormal energy metabolism and muscle biopsy findings]. AB - 31P-NMR spectra were obtained from the quadriceps femoris muscle (at rest and after aerobic exercise) of the 7 cases of mitochondrial myopathies (2 cases of mitochondrial encephalomyopathy and lactic acidosis(MELA), 1 case of myoclonus epilepsy with regged red fiber, 1 case of Kearns-Sayre syndrome, 3 cases of progressive external ophthalmoplegia), using superconducting whole body MR (Magnetom, Siemens). One case showed abnormally low Pcr/Pi ratio in the resting state. An aerobic exercise using ergometer was performed on the other 6 patients. Three of them demonstrated significant reduction and delayed recovery of the Pcr/Pi ratio after exercise. This reduction was not detected in the control subjects. Histological studies of biopsied muscles revealed ragged red fibers in all the cases, the number varies, however, from 0.5 to 15.3 per cent of the total fibers. Abnormalities in the Pcr/Pi ratio of phosphorus spectra, in resting state or after exercise, tend to be observed in patients showing abundant ragged red fibers. Focal cytochrome c oxidase deficiency with relatively small amount of ragged red fibers (less than 10 per cent of the total fibers) was histologically noted in five of our patients, excluding 2 MELA patients. Biochemical assay of mitochondria enzyme was normal. It has been assumed that these patients have no primary defect in energy metabolism and the occasionally observed cytochrome c oxidase deficient fibers are non-specific findings probably caused by some devastating process occurring in these fibers. However, our present studies revealed abnormal reduction and delayed restoration of the Pcr/Pi ratio in 2 out of 5 focal cytochrome c oxidase deficiency cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546708 TI - Estradiol and catecholestradiols as possible genotoxic carcinogens. AB - Estradiol and other estrogens are not classified as genotoxic carcinogens, but rather as tumor promoters in the multistage process of carcinogenesis. This study is a reexamination of the carcinogenic status of estradiol and the catecholestradiols (2-OHE2 and 4-OHE2) with the recently developed bacterial assays for genotoxic carcinogens, the Chromotest. The bacterial kits revealed estradiol and catecholestradiols as biphasic and potential genotoxic carcinogens with the following SOS inducing potency values: E2 43,265 (SD 8,300); 2-OHE2 30,153 (SD 2,500), and 4-OHE2 68,939 (SD 4,500). The differences between these values are statistically highly significant (p less than 0.0005). These results were confirmed by studies on Escherichia coli, which showed an increase in cell number and a stimulation of DNA content in the presence of the estrogens. It is therefore concluded that estradiol and the catecholestradiols are possible genotoxic carcinogens which probably act as tumor inducers rather than tumor promoters. PMID- 2546707 TI - Lipid methylation, hormone action and compensatory hypertrophy in renal cortical tubules. AB - Lipid methylation has been studied in homogenized dog kidney cortical tubules. At pH 9, using S-adenosyl-L-methionine as methyl donor, most of the methyl groups appeared incorporated into phosphatidylcholine, the activity showing an apparent Km of 16 microM. This enzymatic activity was unchanged by the presence of the divalent cations Ca2+ or Mg2+, cAMP or cGMP. In addition, lipid methylation was also unchanged after treatment of intact tubules with angiotensin II, parathyroid hormone or vasopressin. An increased phosphatidylcholine synthesis was observed in the remnant kidney cortical tubules after uninephrectomy through an activation of phosphocholine transferase without detectable modification of lipid methylation. These findings suggest that lipid methylation is not regulated by these biochemical or functional stimuli tested in canine renal cortical tubules. PMID- 2546709 TI - [Magnetic resonance tomography of the parotid gland: plain diagnosis and Gd DTPA]. AB - Pathological lesions of the parotid gland were examined comparatively with different examination sequences both plain and with the contrast medium Gd-DTPA. There were 36 benign lesions (parotitis, Sjogren's syndrome, adenoma, etc.) and 24 malignant tumours (squamous cell carcinoma, adenocarcinoma, adenoid cystic carcinoma etc.) Examinations were carried out at 1.0 T with long and short spin echo sequences in transverse and frontal layer orientation before and after application of Gd-DTPA as contrast medium. In the patients suffering from parotitis the best results were obtained with plain T1 and T2 sequences; the contrast medium Gd-DTPA remained without superior diagnostic relevance. However, in Sjogren's syndrome (myoepithelial sialadenitis) administration of the contrast medium always yielded a characteristic honeycomblike pattern. In benign and malignant space-occupying growths MRI supplied additional diagnostic information with Gd-DTPA in respect of defining the tumour borderlines and paths of infiltration. MRI is now a significant diagnostic tool in inflammatory and tumorous lesions of the parotid gland. PMID- 2546711 TI - Parotid metastases from carcinoma of the breast. AB - Two patients are described in whom a parotid metastasis occurred 10 years after initial treatment for carcinoma of the breast. The parotid is a very unusual site of metastasis and in both patients it was only one detectable. Parotidectomy with post-operative radiotherapy is advocated in this situation not only to obtain local tumour control but to exclude a primary parotid tumour. PMID- 2546710 TI - Computed tomography of hepatic oil embolism following lymphangiography. AB - Computed tomographic (CT) findings in two cases of hepatic oil embolism following lymphangiography are described. Both patients had lymphatic obstruction and opacification of the liver due to collateral flow of lymphangiographic contrast. Computed tomography of the liver showed a distinctive branching pattern due to the intrahepatic ethiodol. PMID- 2546712 TI - Evaluation of assay systems for the detection of rotavirus in stool specimens. AB - We tested 41 rotavirus positive and 42 negative specimens as determined by electron microscopy. The assays systems used were an indirect NIH-ELISA, Meritec Rotavirus, Virogen Rotatest, and Rotazyme II. Meritec and Virogen (latex agglutination assays) were the most sensitive tests, detecting 95% of the positive specimens. The NIH-ELISA detected 81% and Rotazyme detected 63%. Rotazyme was the most specific (100%), followed by the NIH-ELISA (95%) and the two latex agglutination systems (91%). To determine the level of rotavirus detection, we tested three systems against serial two-fold dilutions of ten positive stools. The NIH-ELISA detected rotavirus in an average dilution of 1:723. Rotazyme detected rotavirus in an average stool dilution of 1:366, and Meritec showed an average of 1:164. Rotavirus strains representing serotypes 1-4 were also tested. Meritec was able to detect all four serotypes. Virogen did not react with serotype 2 strains. The NIH-ELISA and rotazyme were unable to detect serotype 3. These data suggest that some latex agglutination assays may be a useful alternative to ELISAs in the clinical laboratory. PMID- 2546713 TI - The phospholipid and fatty acid compositions of seal platelets: a comparison with human platelets. AB - 1. Platelet phospholipid compositions were studied in four species of phocid seals consuming herring or herring and shrimp and in human subjects consuming a normal mixed diet. 2. There were no major differences in platelet phospholipid, cholesterol and protein levels between different species of seal nor between seals and human subjects, nor in the relative abundance of the individual types of phospholipid. 3. The seal platelet phospholipids (phosphatidylcholine (PC) and phosphatidylethanolamine (PE), were greatly enriched in the omega 3 fatty acid, eicosapentaenoic acid (EPA) and depressed in arachidonic acid (AA) relative to the corresponding human platelet phospholipids. 4. Much less accumulation of EPA in phosphatidylserine (PS) and phosphatidylinositol (PI) was found. 5. The EPA contents of the individual seal platelet phospholipids exhibited considerable differences (including EPA discrimination from PI) but gave patterns which were generally similar to those reported for human volunteers consuming fish/fish oils enriched in EPA. 6. These results suggest that the seal platelet may be a useful model for studying the metabolism and function of the omega 3 fatty acids, such as EPA, in relation to platelet reactivity, phospholipid turnover and the formation of AA- and EPA-derived eicosanoids. PMID- 2546714 TI - Effects of repeated androgen treatments on metabolism and nuclear binding of androgen in the infant murine submandibular gland. AB - 1. Androgen responsiveness of esteropeptidase of the murine submandibular gland developed rapidly in normal males compared with in normal females and castrated males. 2. Repeated treatments of infant mice of both sexes with testosterone (T), 5 alpha-dihydrotestosterone (DHT) or 5 alpha-androstane-3 alpha, 17 beta-diol increased androgen responsiveness of this enzyme, but did not affect those of 5 alpha-reductase and 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSDase; androgen metabolizing enzymes) of the gland. 3. Exchange assay of nuclear androgen receptor using 3H-DHT showed that in both sexes, amounts of binding in animals pretreated with T were higher than those in animals pretreated with sesame oil. 4. These results suggest that there is parallelism between the androgen responses and amounts of nuclear androgen binding, not androgen responses of 5 alpha-reductase and 3 alpha-HSDase. PMID- 2546715 TI - Increased survival after massive thermal injuries in adults: preliminary report using artificial skin. AB - Survival of major burn injuries has improved markedly from an expected survival of 10% to 20% in both children and adults to an expected survival of 60% in children with burns greater than 70% BSA. Increased survival for adults after similar burn injuries has been less dramatic than in children because of the profound influences of advancing age and the coexistent processes of aging upon survival after a major injury. Consecutive admissions of patients with massive burn injuries (greater than or equal to 70% BSA) to the Massachusetts General Hospital Adult Burn Unit from 1974 to 1986 were analyzed statistically using univariate and multiple logistic regression analysis to identify factors associated with survival and to identify patient characteristics associated with increases in expected survival. Survival of adults with massive burn injuries has improved markedly, from 24% of adults admitted to the Adult Burn Center in 1974 75 to 48% of adults admitted in 1984-86. An increased likelihood of survival was shown to be associated with the use of artificial skin. Improved survival rates in these massive burn injuries were attributed to multiple factors including wound management of prompt eschar excision and immediate wound closure. PMID- 2546717 TI - Glomerulopathy in renal allografts from patients with and without active Cytomegalovirus infection. AB - Cytomegalovirus (CMV) infection may affect renal graft survival, and a distinctive diffuse glomerulopathy has been described in renal grafts from patients with an active CMV infection. Fifty renal graft biopsies taken 30 to 180 days post-transplant from patients with primary, reactivated, inactive and no CMV infection were examined retrospectively for evidence of this glomerulopathy without knowledge of CMV status. The lesion was found in seven of 30 patients with reactivated CMV infection, one of 12 with primary infection, two of two with no evidence of active infection but evidence of previous infection, and in four of six with no evidence of past or present infection. The differences were not statistically significant. We concluded that the previously described distinctive diffuse glomerulopathy is not specifically associated with CMV infection, but may be a sign of graft rejection. PMID- 2546716 TI - Herpes simplex virus infection complicating amniotic band syndrome in the newborn. AB - Amniotic band syndrome, which is frequently seen in association with prolonged rupture of membranes, is due to the amnion forming fibrous strands that encircle fetal parts. This causes compression and necrosis of the underlying skin. We report a case of herpes simplex virus infection localized to these lesions. PMID- 2546718 TI - Treatment of tracheobronchial granular cell myoblastomas with endoscopic bipolar cautery. AB - Granular cell myoblastomas of the trachea and right upper lobe bronchus were discovered incidentally during therapeutic bronchoscopy. Because of their propensity to cause airway compromise and distal atelectasis, ablation of both lesions was undertaken. This is the first reported case of bipolar cautery of GCM through a flexible fiberoptic bronchoscope. Small tumor size and lack of atelectasis permitted utilization of this technique. Long-term follow-up is necessary to compare this therapy with other nonresectional therapies. PMID- 2546719 TI - Effectiveness of chemoembolization therapy for metastatic right atrial tumor thrombus associated with hepatocellular carcinoma. AB - A rare case of hepatocellular carcinoma (HCC) was complicated by metastatic right atrial tumor thrombus (RATT), which diminished in size on echocardiograms and showed necrotic change on computed tomography (CT) scans after chemoembolization therapy. PMID- 2546720 TI - Determination of total and segmental colonic transit time in constipated patients. Results in 91 patients with a new simplified method. AB - Ninety-one patients with idiopathic constipation had segmental colonic transit studied with radiopaque markers using a new simplified technique to determine frequency and type of colonic transit time (CTT) abnormalities and to determine the utility of this test in planning therapy. Colonic transit studies defined four groups: normal CTT (N = 49), right colonic stasis (N = 16), outlet obstruction (N = 12), and isolated left colonic stasis (N = 14). Right colonic stasis and outlet obstruction were associated with frequent use of digital pressure to assist defecation. Right colonic stasis was characterized by a low stool frequency (less than 3 per week) in 93% of cases and failure to respond to bran therapy. Outlet obstruction also showed a poor response to bran therapy but weekly stool frequency was higher than 3 in 46% of cases. Normal colonic transit time and isolated left colonic stasis were characterized by a normal stool frequency (5.8 +/- 0.05 and 4.2 +/- 0.1, respectively) and clinical help with the use of bran treatment (72 and 64%, respectively). Our study suggested that patients who complain of idiopathic constipation represent a heterogenous group of disorders. Segmental CTT determination is a simple, useful, and noninvasive test of patients with constipation. PMID- 2546722 TI - [The effect of membrane protein methylation processes on the binding of [14C] GABA by nerve endings in the brain of white rats]. PMID- 2546721 TI - Augmentation of cysteamine-induced ulceration of rat duodenum by systemically administered gamma-aminobutyric acid (GABA). AB - Subcutaneous administration of a single dose of gamma-aminobutyric acid (GABA, 10 mg/100 g) in conjunction with a pretreatment dose of aminooxyacetic acid (AOAA 2.5 mg/100 g subcutaneously) to prevent the degradation of GABA, significantly augmented the incidence and intensity of cysteamine HCl-induced duodenal ulceration in rats. This effect of GABA could be reduced by the GABA receptor antagonist, bicuculline (30 micrograms/100 g subcutaneously). These results suggest peripheral GABA receptors can modulate cysteamine HCl-induced duodenal ulcer. PMID- 2546723 TI - [Structure of a potential regulatory area of a gene from the family of the human Na+, K+-ATPase alpha-subunit genes]. PMID- 2546724 TI - [Selective resistance of human neuroblastoma cells to a vaccine strain of poliomyelitis virus]. PMID- 2546725 TI - [The effect of calcium ions on the deactivation of the calcium current in nerve cells of Helix pomatia]. PMID- 2546726 TI - [Infection in malignant blood disorders]. PMID- 2546727 TI - [The causes of leukemia]. PMID- 2546728 TI - [Effect of allogeneic tumor extracts on the development in calves of an experimental infection by the bovine leukemia virus]. AB - The allogenic tumour extracts were studied as immunogens for the prevention of cattle infection with BLV. Immunization did not induce both the production of antibodies against the virus and spontaneous viral infection. Experimental infection of immunized animals with living leukocytes of a leukemic donor caused a BLV infection in 100% of cases. PMID- 2546730 TI - [The level of antibodies to Epstein-Barr virus antigens in the blood serum of the inhabitants of southern Siberia and the Far East]. AB - Antibodies to Epstein-Barr virus have been detected in sera from 1538 healthy adult persons living in the south of Siberia and Far East. It has been shown that the virus infectivity varied from 77% in Russians of the Yakutsk ASSR to 100% in Orochi and Russians residing in the territory of the Gorno-Altai Autonomous Province. Antibody levels to the Epstein-Barr virus for inhabitants of the south regions were higher than those for inhabitants of the north regions. PMID- 2546729 TI - [The role of protein kinase C in tumor growth]. AB - A review of recent papers dealing with the role of protein kinase C (PK C) in regulation of normal and tumour cell proliferation contains the data on the interaction of PK C with the tumour promoters and oncoproteins as well as the data which characterize changes of the PK C activity in tumour cells and during carcinogenesis. Probable mechanisms of the PK C influence on the functioning of the growth factors receptors are discussed. It is proposed that one of the PK C functions which is substantial for induction of the mitogenic signal is the polyamine synthesis stimulation. Participation of PK C in the tumour promotion may play a key role in multistep processes of the carcinogenesis. PMID- 2546731 TI - [Effect of the prostaglandin E produced by tumor cells on the enzymatic activity of adenosine metabolism and on human natural killer cytotoxic activity]. AB - Adenosine deaminase (ADA) and 5'-nucleotidase (5'-N) activity and cytotoxic activity (CTA) of natural killer cells (NK-cells) isolated from human peripheral blood were studied. These cells were affected by culture medium obtained after the contact of tumour cells with NK-cells, the degree of malignancy in these cells being different. It has been established that pretreatment of NK-cells with culture medium from highly malignant cells and containing PGE, causes a sharp inhibition of their CTA, which is accompanied by an essential decrease of the ADA activity and an increase of the 5'-N activity in these cells as compared with the control. PMID- 2546732 TI - Luteinizing hormone-releasing hormone-binding sites in the rat thymus: characteristics and biological function. AB - The present study was designed to explore the effects of LHRH and its agonists on immune system function. As a first step, to identify a putative site of action, the very potent and stable LHRH agonist (LHRH-A), [D-Ser(TBU6)] des-Gly10-LHRH ethylamide (buserelin), was used as an iodinated ligand to characterize LHRH receptors in a membrane preparation of rat thymus, a key organ of the immune system. The effects of LHRH and LHRH-A were then investigated on the proliferative capacity of rat thymocytes exposed in vitro to a mitogen and on ornithine decarboxylase specific activity. In addition, to determine whether LHRH A treatment in vivo might directly influence thymic function, we treated hypophysectomized (hypox) rats with a moderately high dose of LHRH-A for a period of 2 weeks, and thymocyte mitogenic capacity, thymus weight, and the histological and functional appearance of the thymus were then assessed. Specific binding of LHRH-A to rat thymic membrane preparations is a saturable process, depending on both time and temperature of incubation, but differs markedly from binding to the rat pituitary or ovarian LHRH receptor in its low binding affinity. Binding is optimal in the absence of chelating agents (EDTA) or divalent metal ions, and increases linearly with increasing protein concentration. Binding is specific for LHRH, LHRH-A, and antagonists. Both the C-terminal amide and N-terminal regions of the LHRH molecule were required for binding, and amino acid substitutions at position 6 markedly enhanced and at position 8 markedly reduced binding potencies in rat thymic tissue. A number of peptides, proteins, and other agents had no effect on the specific binding of LHRH-A to thymic membrane preparations. The binding affinity (Ka) of the membrane receptor of the rat thymus for the LHRH superagonist buserelin was 8.4 x 10(8) M-1, while a higher binding affinity (Ka = 2.8 x 10(9) M-1) was calculated for the ovarian LHRH-binding site. Preincubation of rat thymocytes with LHRH-A for 20 h induced a significant dose-dependent increase in the proliferative response to the mitogen Concanavalin-A, monitored by [3H]thymidine incorporation. Using native LHRH, it was also possible to elicit stimulatory effects on the same parameter, although much higher concentrations were required than with LHRH-A. Furthermore, simultaneous addition of a LHRH antagonist, abolished the LHRH effect on thymocytes. Ornithine decarboxylase specific activity under lectin stimulation was also significantly increased by LHRH-A in cultures of rat thymocytes.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2546733 TI - Luteinizing hormone-releasing hormone (LHRH) agonist restoration of age associated decline of thymus weight, thymic LHRH receptors, and thymocyte proliferative capacity. AB - The presence of specific LHRH-binding sites within the rat thymus gland and the ability of LHRH and its agonistic and antagonistic analogs to directly modulate thymus function prompted us to study the possible changes in the number of thymic LHRH-binding sites during aging-induced physiological immunosenescence. Moreover, the effects of chronic treatment of aging rats with a potent LHRH agonist (LHRH A) on thymic LHRH receptors, thymus weight and histology, as well as thymocyte proliferative capacity were assessed. For comparison, the effects of castration on the same parameters was also investigated. The process of aging is accompanied by a sharp reduction in LHRH-A-binding sites within the thymus gland of both female and male rats. Starting at 7 months of age, a 50% decrease in thymic LHRH A binding was followed, at 11-13 months of age, by a nearly 65% inhibition of receptor numbers. In 16- to 19-month-old rats, LHRH-A binding was almost completely lost. Thymus weight was 30% reduced in 7-month-old animals, while a 50% reduction in thymic size was reached at 11 months of age in males and 13 months in female rats. A further decrease in thymic mass was observed at 16 and 19 months. Chronic (45-day) treatment of aging (15-16 months old) female and male rates with the potent LHRH-A, [D-Trp6,Des-Gly10]LHRH-N-ethylamide, reversed the age-related decreases in both thymus weight and thymic LHRH-binding sites. Similarly, surgical removal of testicular hormones by castration restored thymus weight and increased LHRH-A binding in the thymus of aged rats. While thymus histology in 3-month-old rats was characterized by a clear demarcation of cortical and medullary regions, only thymic remnants were present in 16- to 17 month-old animals. Castration of old rats resulted in a partial restoration of thymic structure, while chronic treatment of aging rats with the LHRH-A produced a homogeneous organization of both cortical and medullary compartments accompanied by a marked increase in the width of the cortical layer, densely packed with lymphocytes. While the process of aging was accompanied by an almost complete loss of the proliferative response of thymocytes to optimal concentrations of the mitogen Concanavalin-A, thymocyte cultures from old rats treated with LHRH-A or from castrated animals, displayed significantly greater proliferative responses. Furthermore, the combination of both manipulations resulted in a further significant increase in thymocyte proliferative capacity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2546734 TI - The alkyl-ether phospholipid platelet-activating factor is a stimulator of the hypothalamic-pituitary-adrenal axis in the rat. AB - Platelet-activating factor (PAF) is a naturally occurring alkyl-ether phospholipid which serves as an extracellular mediator in various cellular processes. Here we examined the effects of PAF on the activity of the hypothalamic-pituitary-adrenal axis in vivo and in vitro. PAF injected iv to rats (125, 250, and 500 ng/100 g BW) caused significant stimulation of pituitary ACTH and adrenal corticosterone secretion. The peak of PAF effect was recorded 10 min after the injection. Intraperitoneal injection of the PAF receptor antagonist BN 52021 prevented the ACTH-releasing effect of PAF. In explanted rat hypothalami maintained viable in vitro, PAF stimulated immunoreactive CRH secretion in a bell shaped dose-response fashion. The maximal stimulatory effect occurred at the concentration of 10 nM. Higher concentrations appeared to cause desensitization. Alprazolam (1 microM) and BN 52021 (1 microM), two structurally different PAF receptor antagonists, inhibited this effect. The inhibitors of arachidonic acid metabolism, indomethacin, eicosatetraynoic acid, and the calcium channel blocker verapamil, inhibited PAF-stimulated CRH secretion, suggesting mediation by Ca2+ influx and phospholipase-A2 activation. In addition, we found that 1 nM PAF weakly stimulated ACTH secretion by dispersed rat pituicytes. This stimulatory effect of PAF was also inhibited by the receptor antagonists alprazolam and BN 52021. Our data suggest that PAF plays a role in the activation of the hypothalamic-pituitary-adrenal axis and glucocorticoid secretion and can perhaps serve as a mediator in the interactions of the immune system with the central nervous system. PMID- 2546735 TI - Stimulation of glucose transport in rat cardiac myocytes by guanosine 3',5' monophosphate. AB - Glucose transport in isolated rat cardiomyocytes is stimulated by insulin, catecholamines, and anoxia approximately 2- to 3-fold over basal rates. The molecular mechanisms controlling these responses are unknown. In our search for possible cellular mediators of glucose transport stimulation, we examined the effects of a number of nucleotides on 3-O-methylglucose transport in heart cells. The nucleotides and/or permeable analogs (monosuccinyl, 8-bromo, and dibutyryl derivatives) included cUMP, cIMP, cCMP, cAMP, and cGMP at concentrations ranging from 10 nM to 1 mM. Of all the nucleotides tested only cGMP analogs induced a significant stimulation of transport at concentrations as low as 100 nM. This effect was observed in both the 8-bromo- and dibutyryl derivatives and with 1 mM cGMP itself. The effect was concentration dependent for both analogs and produced a maximal response equivalent to that of 100 nM insulin. This insulinomimetic effect of cGMP was examined in more detail in order to evaluate its role as a potential mediator of this response. Agents that are known to stimulate guanylate cyclase in the heart produced a clear stimulation of transport when added to cardiomyocytes. These include insulin, aminophylline, histamine, beta-estradiol, and biotin-nitrophenyl ester. Methylene blue, an inhibitor of guanylate cyclase, blocked the insulin response when added to cells before insulin, but was ineffective when added after insulin. In addition, agents that raise intracellular cGMP levels by inhibiting cyclic nucleotide phosphodiesterases were also examined for effects on glucose transport. Out of several phosphodiesterase inhibitors tested, only Zaprinast (which selectively increases cGMP in heart) stimulated transport in a concentration-dependent manner to within 80% of the maximal insulin effect. These results are consistent with the notion that cGMP may be involved in glucose transport stimulation. PMID- 2546736 TI - Renal and adrenal adenosine 3',5'-monophosphate production and corticosteroid secretion in response to synthetic chicken parathyroid hormone-(1-34). AB - The activity of synthetic chicken (c) PTH-(1-34) amide was tested in dispersed chicken and rat kidney and adrenocortical cells. In the adrenal cells the effect of intact cPTH was also evaluated. In chicken kidney cells, the time- and dose response patterns of cAMP production were similar for cPTH-(1-34) amide and human (h) PTH-(1-34), whereas rat kidney cells were considerably more sensitive to hPTH (1-34) than to cPTH-(1-34) amide. The agonist effects of both hPTH-(1-34) and cPTH-(1-34) amide in kidney cells were inhibited by the bovine PTH-(3-34) analog. In chicken adrenocortical cells, cPTH-(1-34) amide stimulated cAMP production and steroid secretion. This action of the peptide was inhibited by bovine PTH-(3-34) and hPTH-(1-34), which by themselves showed no agonist effects. The maximal response of steroid secretion to cPTH-(1-34) amide was significantly lower than that to ACTH, but intact cPTH (supplied as a semipurified parathyroid extract) stimulated steriodogenesis to the same extent as ACTH. In rat adrenocortical cells, intact cPTH stimulated both cAMP formation and steriodogenesis, but cPTH (1-34) amine showed no agonist effect. The action of the intact hormone in the rat adrenal could be inhibited by cPTH-(1-34) amide. The present results demonstrate the interaction of cPTH-(1-34) with kidney and adrenocortical cells of either chicken or rat. The cAMP and steroidogenic responses of the adrenocortical cells to PTH appear to be dependent (completely in the rat and partially in the chicken) on some sequence beyond the 1-34 region. PMID- 2546737 TI - Pulsatile release of gonadotropin-releasing hormone from hypothalamic explants is restrained by blockade of N-methyl-D,L-aspartate receptors. AB - We have shown previously that N-methyl-D,L-aspartate (NMDA) and kainate, two neuroexcitatory amino acids acting through distinct receptors, may induce the release of GnRH from hypothalamic explants. However, that effect could have no physiological significance, since very high concentrations (50 mM) of NMDA and kainate were required. Here, using agents blocking the activation of receptors to neuroexcitatory amino acids, we evaluated their possible physiological involvement in the pulsatile release of GnRH from the hypothalamus of 50-day-old male rats in vitro. In control conditions (10 nM glycine and 1 mM mg2+), the release of GnRH in 7.5-min fractions collected for 2-4 h showed an obvious pulsatile pattern. The mean (+/- 1 SD) interval between pulses, identified by PULSAR program, was 34.3 +/- 11.4 min. The stimulation of GnRH release by NMDA (50 mM) added to the medium for 7.5 min could be blocked reversibly in the presence of MK-801 (100 microM) using medium without glycine or enriched with Mg2+ (2 mM). The endogenous pulses of GnRH secretion were abolished in the presence of MK-801 or using increased Mg2+ concentrations as well as in the absence of glycine. In contrast, pulsatile release of GnRH was not affected in the presence of 6,7-dinitroquinoxaline-2,3-dione (0.1 mM), a selective inhibitor of kainate and quisqualate receptors which suppressed the increase in GnRH release induced by kainate (50 mM) without affecting the response to NMDA. These data indicate that the physiological mechanism of pulsatile GnRH secretion in the hypothalamus may involve endogenous neuroexcitatory factors acting through NMDA sensitive receptors. PMID- 2546739 TI - Stimulation of glucose production by activin-A in isolated rat hepatocytes. AB - The effect of activin-A on glycogenolysis was studied in isolated rat hepatocytes. Activin-A stimulated glucose output in hepatocytes in a dose dependent manner. The maximal effect of the glycogenolytic action of activin-A, which was about 50% of the glucagon action, was obtained at 10(-9) M. When 10(-9) M activin-A and 5 x 10(-9) M glucagon were added simultaneously, the actions of these two agents were additive. In contrast, there was no additivity when 10(-9) M activin-A and 10(-8) M angiotensin-II were added. Activin-A did not increase cAMP at any doses tested, but induced a rapid increase in cytoplasmic free calcium concentration. Activin-A increased the cytoplasmic free calcium concentration even in the presence of 1 microM extracellular calcium, suggesting that activin-A caused calcium release from an intracellular calcium pool(s). The internal calcium pool affected by activin-A appeared to be the same as that affected by either angiotensin-II or vasopressin. When [3H] inositol-labeled hepatocytes were incubated with activin-A, radioactivity in the inositol trisphosphate fraction was rapidly increased. These results indicate that activin A acts on rat hepatocytes and stimulates glycogenolysis by activating the calcium messenger system. PMID- 2546738 TI - Adrenocorticotropin(1-10) and -(11-24) promote adrenal steroidogenesis by different mechanisms. AB - ACTH1-10 and ACTH11-24 each elicit cortisol secretion submaximally in freshly dispersed or cultured beef adrenal cortical cells. The combination of ACTH1-10 and ACTH11-24 promotes cortisol release to the maximal level elicited by ACTH1 24. Maximal cortisol release by ACTH11-24, but not by ACTH1-24 or ACTH1-10, was enhanced by forskolin. The calcium channel blockers nifedipine and verapamil inhibited cortisol release by ACTH1-10, ACTH1-24 or ACTH11-24, suggesting calcium influx to be essential for steroid secretion regardless of the secretagogue. Vanadium, in a dose-dependent manner, inhibited cortisol secretion elicited by ACTH1-24 and ACTH1-10 but not that caused by ACTH11-24. These results suggest that there are at least two receptors mediating ACTH1-24-dependent steroid secretion. One class of receptor recognizes ACTH1-10 but not ACTH11-24 and is linked to the cAMP messenger pathway. PMID- 2546740 TI - Uterine insulin-like growth factor-I receptors: regulation by estrogen and variation throughout the estrous cycle. AB - This study was undertaken to identify uterine insulin-like growth factor-I (IGF I) receptors and examine the regulation of these receptors throughout the estrous cycle and after 17 beta-estradiol (E2) administration to immature rats. We have demonstrated type I IGF receptors in crude uterine membranes by binding and cross linking experiments. The characteristics of the uterine IGF-I receptor are similar to those reported for other tissues, with a high affinity component (Kd = 0.12 nM; binding capacity = 0.028 pmol/mg protein) and a low affinity component (Kd = 0.98 nM; binding capacity = 0.041 pmol/mg protein). Autoradiographic visualization of [125I]IGF-I binding to uterine sections localized the IGF-I receptors to the smooth muscle cells of the myometrial layer of the uterus, with a higher density of IGF-I receptors in the outer longitudinal layer than in the inner circular layer. In immature rats administration of E2 significantly increased total [125I]IGF-I binding per uterus as early as 6 h after E2 injection. Although [125I]IGF-I binding was significantly increased per mg DNA, because of a more marked increase in membrane protein after E2, [125I]IGF-I binding, when expressed per mg membrane protein, decreased. This change in [125I]IGF-I binding resulted from a change in receptor number with no change in receptor affinity. In mature cycling rats, the proestrous uteri showed the lowest level of [125I]IGF-I binding per mg membrane protein, although because of the greater yield of protein from proestrous uteri, the total [125I]IGF-I-binding capacity of these uteri were greater than that of uteri from other stages of the estrous cycle. The lowest [125I]IGF-I binding was seen in the diestrous uteri. These studies demonstrate the presence of authentic type I IGF-I receptors in the rat uterus localized predominantly to the myometrial smooth muscle cells. In addition, E2 appears to regulate the uterine IGF-I receptor in the immature rat, and the cyclical changes in the mature rat are consistent with a role of E2 in regulation of this receptor in vivo. PMID- 2546741 TI - Combined effects of dexamethasone and 1,25-dihydroxyvitamin D3 on parathyroid hormone secretion in cultured bovine parathyroid cells. AB - The present studies investigate the effects of glucocorticoids on the function of the parathyroid glands using primary cultures of bovine parathyroid cells. Treatment of parathyroid cell cultures with dexamethasone for 48 h caused a dose dependent stimulation of PTH secretion. The minimal concentration of dexamethasone required for a significant stimulation of PTH secretion was 0.1 nM. The stimulatory effect of dexamethasone on the secretion of PTH was found within 12 h of treatment with 100 nM dexamethasone. The steroids deoxycorticosterone and cortexolone, which do not have glucocorticoid activity were without effect of PTH secretion. Since glucocorticoids may modulate the effects of 1,25 dihydroxyvitamin D3 [1,25(OH)2D3] in other tissues, additional studies were performed to evaluate the interactions of glucocorticoids and 1,25-(OH)2D3. Addition of 1,25-(OH)2D3 to parathyroid cell cultures for 48 h significantly suppressed PTH secretion. In the presence of dexamethasone, however, 1,25-(OH)2D3 also significantly decreased PTH secretion, although it did not reduce PTH secretion to control levels. The treatment of parathyroid cell cultures with 100 nM dexamethasone did not affect the parathyroid cell content of 1,25-(OH)2D3 receptors. In summary, these studies indicate that glucocorticoids significantly increase the secretion of PTH in vitro. This stimulatory effect can be inhibited by 1,25-(OH)2D3. The parathyroid gland is an additional site of physiological antagonism of glucocorticoids and 1,25-(OH)2D3. PMID- 2546742 TI - Transferrin in FRTL5 cells: regulation of its receptor by mitogenic agents and its role in growth. AB - Transferrin, a serum iron-binding protein, delivers iron to the cell after binding to specific receptors on the cell surface and is an important component of culture medium for virtually all cell lines, including the FRTL5 line of rat thyroid follicular cells. Therefore, we undertook studies in FRTL5 cells to examine the regulation of the transferrin receptor, the effects of transferrin on growth and differentiated functions, and the interactions of transferrin with several mitogenic pathways. FRTL5 cells possess one class of saturable transferrin receptors (Ka, 0.7 x 10(9) M-1). Binding of 125I-labeled transferrin was highest in actively growing cells and declined progressively, reaching minimal values when confluence was achieved. Removal of transferrin from culture medium caused a rapid increase in transferrin binding. TSH, acting within 5 min, induced a modest increase in transferrin binding, due to a cycloheximide resistant increase in binding sites. Binding of transferrin after a 24-h incubation was also increased by other mitogenic agents, (Bu)2cAMP, forskolin (FK), insulin, insulin-like growth factor-I (IGF-I), and the phorbol ester TPA. Transferrin alone stimulated growth only minimally, but enhanced the mitogenic effect of TSH, (Bu)2cAMP, and FK, all of which act through the cAMP pathway. In contrast, transferrin did not alter the cAMP-independent mitogenic effects of insulin and IGF-I. Transferrin did not affect TSH-induced cAMP generation. Desferoxamine, an iron chelator, inhibited the mitogenic effects of all of the agents tested. Desferoxamine had no significant effect on TSH-induced cAMP accumulation. We conclude that FRTL5 cells contain saturable receptors for transferrin whose abundance varies with the rate of cell replication. Transferrin down-regulates its own receptors, while stimulation of growth by various mitogens is accompanied by increased binding of transferrin. Transferrin enhances the mitogenic effect of the cAMP-dependent mitogens, TSH, (Bu)2cAMP, and FK, without modifying basal or stimulated cAMP generation. In contrast, transferrin fails to affect the mitogenic responses to IGF-I and insulin, which are cAMP independent. Iron is required for the mitogenic response to various mitogens, especially those that are cAMP dependent. PMID- 2546743 TI - The role of Ca2+ and calmodulin in the regulation of atrial natriuretic peptide stimulated guanosine 3',5'-cyclic monophosphate accumulation by isolated mouse Leydig cells. AB - We have investigated the role of Ca2+ and calmodulin in the stimulation of cGMP formation by mouse Leydig cells in response to rat atriopeptin-II (rAP-II). Removal of extracellular Ca2+ had no influence on the levels of cGMP accumulated by the cells stimulated with rAP-II. The amounts of testosterone produced by unstimulated and rAP-II-stimulated cells were, however, reduced by 50% in the absence of Ca2+ from the incubation medium. Addition of ionomycin to the Leydig cells led to a dose-related inhibition of rAP-II-stimulated cGMP formation, but the basal cGMP level was not affected. These experiments were carried out in the presence of a phosphodiesterase inhibitor. The inhibitory effect of ionomycin was absolutely dependent upon the presence of Ca2+ in the medium. The guanylate cyclase activity required the presence of a cation, and Mn2+, Mg2+, or Ca2+ could function as the required cation. There was no direct inhibition of the cyclase activity by Ca2+ up to as high a concentration as 8 mM. Furthermore, three structurally unrelated calmodulin antagonists, W7, trifluoperazine, and calmidazolium, but not W5, caused a dose-related inhibition of rAP-II-stimulated cGMP accumulation by the cells. The inhibitory effect of calmodulin antagonists was not exerted directly at the level of guanylate cyclase activity, since the particulate enzyme was not inhibited by any of these drugs. We conclude, therefore, that extracellular Ca2+ is not essential for rAP-II-mediated stimulation of cGMP formation by mouse Leydig cells, at least under the short term incubation conditions used. An excessive ionophoretic influx of Ca2+ into the cells impairs the ability of rAP-II to stimulate cGMP formation. Therefore, it appears that a finely regulated level of intracellular Ca2+ is required for optimal activation of atrial natriuretic peptide-responsive guanylate cyclase in mouse Leydig cells, and calmodulin plays an important role in this process. PMID- 2546744 TI - Nitroimidazole derivatives inhibit anterior pituitary cell function apparently by a direct effect on the catalytic subunit of the adenylate cyclase holoenzyme. AB - Nitroimidazole derivatives dose-dependently decreased basal and CRF-stimulated ACTH release, basal and GRF-stimulated rat GH release, and basal rat PRL release in primary cultures of rat anterior pituitary cells. In addition, basal and CRF stimulated mRNA coding for the ACTH precursor were reduced after preincubation with the nitroimidazole derivatives. Miconazole, econazole, isoconazole, clotrimazole, and bifonazole had similar or more pronounced effects on anterior pituitary function compared to ketoconazole, whereas metronidazole and etomidate were less effective. The positive correlation between the number of phenylated side-chains or phenolic rings of the imidazole molecule and the efficacy to inhibit activity on pituitary hormone secretion suggests a structure-activity relationship of these compounds. The effects of the nitroimidazole derivatives on anterior pituitary hormone release and biosynthesis were mediated by cAMP. Thus, basal and CRF-, cholera toxin-, and forskolin-stimulated adenylate cyclase activities in rat anterior pituitary cell membranes determined by cAMP formation were suppressed by the nitroimidazole derivatives. Pertussis toxin did not diminish the nitroimidazole derivative effect on cAMP formation. The adenylate cyclase inhibitory effect of these substances was independent of the presence of GTP in the assay system, underlining a direct effect on the catalytic subunit. In addition, basal and forskolin-stimulated cAMP generation in membranes of S49 lymphoma cyc-variants, which lack a functional Gs protein, was efficiently suppressed (by up to 90%) by the nitroimidazole derivatives. In conclusion, ketoconazole and other nitroimidazole derivatives inhibit anterior pituitary hormone synthesis and secretion apparently by a direct effect on the catalytic subunit of the adenylate cyclase system. PMID- 2546745 TI - Acetylcholine stimulates alpha-melanocyte-stimulating hormone release from frog pituitary melanotrophs through activation of muscarinic and nicotinic receptors. AB - The release of alpha MSH from the pars intermedia of amphibians is regulated by multiple factors, including classical neurotransmitters and neuropeptides. In this study we have examined the possible involvement of acetylcholine (ACh) in the regulation of alpha MSH secretion from the pars intermedia of the frog (Rana ridibunda) using the perifusion technique. When intact neurointermediate lobes (NIL) were exposed to graded doses of ACh (3 X 10(-7) to 3 X 10(-4) M), a dose dependent stimulation of alpha MSH release was observed. Repeated administration of ACh (10(-4) M) induced reproducible responses of NIL without any desensitization phenomenon. ACh was also capable of stimulating alpha MSH release from dispersed intermediate lobe cells, indicating that the neurotransmitter exerts its effect by acting directly on frog melanotrophs. Using the monoclonal antibody M-35 against calf muscarinic receptors we have visualized, by the immunofluorescence technique, the presence of muscarinic receptor-like immunoreactivity in the frog pars intermedia. The stimulatory action of ACh was mimicked by both nicotine and muscarine (10(-5) M each). Nicotine-induced stimulation of alpha MSH release was partially abolished by alpha-bungarotoxin (10(-6) M) and hexamethonium (10(-4) M). The stimulatory effect of muscarine was suppressed by atropine and the M1-muscarinic antagonist pirenzepine (10(-5) M), but not by the M2-muscarinic antagonist gallamine. We have investigated the effect of ACh during administration of specific nicotinic and muscarinic antagonists. While hexomethonium or atropine could block only part of the stimulatory effect of ACh, concomitant administration of these antagonists totally abolished the response of NIL to ACh. Finally, the stimulatory effect of ACh was not impaired during prolonged administration of the beta-adrenergic antagonist propranolol. These data show that ACh stimulates in vitro alpha MSH secretion by frog NIL. Our results also indicate that amphibian pars intermedia cells possess two types of cholinergic receptors, an M1-muscarinic receptor sensitive to pirenzepine and nicotinic receptors sensitive to hexamethonium and alpha-bungarotoxin. PMID- 2546746 TI - Highly polarized secretion of inhibin by Sertoli cells in vitro. AB - We have studied the regulation of inhibin secretion by rat Sertoli cells grown on extracellular matrix-impregnated porous filters in a twin chamber assembly. Previous studies have established that rat Sertoli cells cultured under these conditions reproduce the morphological and functional polarization observed in the Sertoli cell in situ. Sertoli cells isolated from 18- to 22-day-old Wistar rats were cultured for up to 8 days with daily changes of fully defined supplemented Eagle's Minimum Essential Medium (MEM). Rat inhibin was measured by RIA and pituitary cell bioassay, and transferrin by RIA. Inhibin measured by immunoassay or bioassay was always readily detectable in the upper, but not the lower, chamber. Inhibin secretion into the upper chamber exhibited a dose dependent stimulation of up to 3.7-fold by ovine FSH, with a medium effective dose of 2.2 micrograms/liter and a constant bio- to immunoreactive ratio (3.6 +/- 0.4). Apically directed secretion accounted for over 80% of inhibit output under basal conditions and over 94% with FSH stimulation. Insulin also stimulated upper chamber inhibin secretion at a high dose (5 mg/liter) but not at lower doses or in conjunction with FSH exposure of Sertoli cells. Testosterone augmented FSH induced stimulation of inhibin secretion, but was ineffective without FSH exposure. In contrast to inhibin secretion, for which FSH is the principal regulator, transferrin secretion by Sertoli cells is more evenly bidirectional (overall mean upper to lower chamber ratio of 1.5) and requires exposure to other stimuli (insulin, retinoic acid, and testosterone) in addition to FSH to achieve maximal secretion. Both submaximal and maximal FSH stimulation of inhibin output were augmented by a phosphodiesterase inhibitor, isobutylmethylxanthine, and these effects were fully reproduced by forskolin, which suggests the involvement of cAMP in the vectorial secretion of inhibin. The marked polarization of Sertoli cell inhibin secretion in vitro could not be explained by restricted transmembrane passage of inhibin. It is, therefore, suggested that the bulk of inhibin secretion by the immature rat Sertoli cell in vivo may be directed primarily into the seminiferous tubular lumen. Thus, in addition to its role in endocrine negative feedback signaling to the pituitary, inhibin may also have important functions in seminiferous tubular function and the support of spermatogenesis. PMID- 2546748 TI - Effects of naloxone on vasopressin secretion in conscious rats: evidence for inhibitory role of endogenous opioid peptides in vasopressin secretion. AB - The effects of naloxone, an opioid antagonist, on arginine vasopressin (AVP) secretion were examined in conscious unrestrained rats under both basal and stimulated conditions. Intravenous injection of naloxone in a dose of 0.1 mg/kg did not significantly affect the basal plasma AVP level. However, 0.5 or 2.5 mg/kg naloxone significantly raised the basal AVP level in euhydrated rats. Naloxone (0.5 mg/kg) significantly enhanced AVP secretion after 72-h water deprivation. However, the enhancement was more prominent in euhydrated rats than in dehydrated rats. Pretreatment with naloxone (0.5 mg/kg) also significantly prolonged AVP secretion induced by intracerebroventricular injection of angiotensin-II (100 ng). Moreover, naloxone (0.5 mg/kg) significantly increased AVP secretion induced by intracerebroventricular injection of carbachol (10 ng). Naloxone (0.5 mg/kg) altered neither basal blood pressure nor the angiotensin-II induced pressor response, but augmented the carbachol-induced pressor response. This suggests that facilitation of AVP secretion by naloxone is not due to a reflex mechanism resulting from decreased blood pressure. These results indicate that endogenous opioid peptides exert a tonic inhibitory control on AVP secretion in rats. PMID- 2546749 TI - Aldosterone-stimulated down-regulation of both type I and type II adrenocorticosteroid receptors in mouse brain is mediated via type I receptors. AB - The concentrations of type I and type II adrenocorticosteroid receptors in brain cytosol obtained from adrenalectomized-ovariectomized female mice were measured with five different assay conditions. Among the five brain regions studied, hippocampus had the highest concentration of type I receptors, whereas cerebral cortex had the highest concentration of type II receptors. The value of properly correcting for dexamethasone cross-binding to type I receptors when type II receptors are being assayed was demonstrated using the type II receptor-selective ligand RU28362. A time-course study revealed a transient up-regulation of both receptor classes in most brain regions after adrenalectomy-ovariectomy, with maximal values achieved 3-5 days postsurgery and a reduction to near-intact levels by 16 days postsurgery. A single sc injection of aldosterone given to adrenalectomized-ovariectomized mice produced a profound down-regulation of type I receptors in hippocampal, cerebral cortex, hypothalamic, brain stem, and cerebellar samples, whereas it down-regulated type II receptors only in hippocampal and cerebral cortical samples. A similar injection of RU28362 failed to down-regulate type I receptors in any brain region, but it did reduce the concentration of type II receptors in all brain regions except cerebellum. The actions of aldosterone appear to be mediated solely through type I receptors, since injections of the type I receptor antagonist RU26752 prevented aldosterone induced down-regulation of both type I and type II receptors, whereas RU26752 had no effect on the down-regulatory actions of RU28362. The ability of aldosterone to down-regulate type I, but not type II, receptors in hypothalamic, brain stem, and cerebellar samples suggests that type I and type II receptors are concentrated in separate populations of cells in these brain regions, whereas in hippocampus and cerebral cortex there is a sufficient degree of colocalization to permit type II receptor down-regulation via the action of aldosterone-type I receptor complexes. We speculate that this action is mediated at least in part at the genomic level by the suppression of type I and type II receptor mRNA synthesis brought about by the interactions of transformed aldosterone-type I receptor complexes with the DNA regulatory elements upstream from the genes for these receptors. PMID- 2546747 TI - Two squamous cell carcinomas not associated with humoral hypercalcemia produce a potent bone resorption-stimulating factor which is interleukin-1 alpha. AB - Conditioned medium (CM) from two squamous cell carcinoma cell lines, SCC-9 and SCC-13, stimulated bone resorption in neonatal mouse calvariae in organ culture. Enhanced bone resorption induced by CM was associated with an increased production of prostaglandin-E2 (PGE2) by the calvariae. Complete inhibition of stimulated PGE2 synthesis by indomethacin only partially inhibited bone resorption-stimulating activity (BRSA) in the CM. Neither SCC-9 nor SCC-13 CM stimulated cAMP production in rat osteosarcoma cells (ROS 17/2.8). The BRSA in CM was completely inhibited by an antibody to interleukin-1 alpha (IL-1 alpha). Fractionation of SCC-9 CM by gel filtration and HPLC ion exchange chromatography revealed a single peak of BRSA and PGE2 synthesis-stimulating activity at 17-20K (termed SCMII). In mouse calvariae, SCMII increased medium Ca2+ and PGE2 in a dose-dependent manner at concentrations from 20 ng protein/ml to a maximum of 500 ng protein/ml. Preincubation of SCMII with antibody to IL-1 alpha completely inhibited SCMII-induced bone resorption. SCMII also enhanced thymocyte proliferation with activity that was equivalent to 353 U/ml IL-1. Antibodies to IL-1 beta and tumor necrosis factor had no effect on SCMII-induced bone resorption. Using specific enzyme-linked immunosorbent assays for IL-1 alpha and IL-1 beta, IL-1 alpha was measured in high concentrations in both crude and partially purified fractions of SCC-9 and SCC-13 CM. In contrast, IL-1 beta was either undetectable or present in amounts below those that stimulate bone resorption. In addition, SCMII did not enhance cAMP production in bone cells. We conclude that the BRSA produced by the two squamous cell carcinoma cell lines SCC 9 and SCC-13 is IL-1 alpha. PMID- 2546750 TI - Characterization of thyroid hormone effects on Na channel synthesis in cultured skeletal myotubes: role of Ca2+. AB - Thyroid hormones (TH) cause an increase in spontaneous electrical activity of cultured rat skeletal myotubes. This activity is associated with tetrodotoxin (TTX)-sensitive Na channels. In addition, the initial effect of TH on Na-K pump synthesis has been shown to be TTX dependent. Accordingly, we have studied effects of TH on expression of TTX-sensitive Na channels in cultured skeletal muscle. Expression of Na channels was determined by measurements of the binding of [3H]saxitoxin (STX). The frequency and rate of rise of spontaneously occurring action potentials, which are related to the density of TTX-sensitive Na channels, were also determined. TH caused dose-dependent increases in Na channels as well as in action potential frequency and rate of rise. The increases were detectable as early as 12 h after treatment with TH was begun, and levels reached a maximum plateau after 36-48 h. The effects of TH were blocked by inhibitors of protein synthesis. Scatchard analysis showed the channels in TH-treated myotubes to have lower affinity for STX than those in control cells. The effect of TH to up regulate Na channels was reduced by growth of the cells in elevated external calcium. In contrast, treatment with TTX or verapamil, which lower cytosolic Ca2+, resulted in a marked increase in the effect of TH over that in control myotubes. Thus, TH appears to regulate Na channels in cultured myotubes by two opposing mechanisms; 1) direct stimulation of Na channel synthesis, and 2) indirect decrease in synthesis mediated by an increase in cytosolic Ca2+. The results indicate that TH may play an important role in developmental expression of Na channels in excitable tissue. PMID- 2546751 TI - Multiple factors influence insulin-like growth factor-I binding to human skin fibroblasts. AB - The binding of [125I]insulin-like growth factor-I ([125I]IGF-I) to human skin fibroblasts (HSF) is regulated by multiple factors. In monolayers of HSF, IGF-I binds to both the type I IGF receptor and IGF-binding proteins (BPs) associated with the cell surface. [125I]IGF-I binding to both of these proteins depends markedly on the sodium chloride concentration of the binding buffer. In monolayers of HSF, replacing 120 mM NaCl with isoosmotic concentrations of sucrose increases binding of [125I]IGF-I by 2- to 6-fold. Enhancement of [125I]IGF-I binding in the absence of sodium chloride is also seen in HSF in suspension, in human erythrocytes, in monolayers of HEP G2 cells and FRTL5 cells, and in membranes prepared from human placentae. Kinetic analysis of [125I]IGF-I binding to HSF monolayers reveals that association rates are increased and dissociation rates are decreased in the absence of sodium chloride. The binding of [125I]alpha IR-3, a monoclonal antibody to the human type I IGF receptor, to monolayers and suspensions of HSF also depends on the sodium ion concentration; it is 5- to 7-fold higher in the absence of sodium chloride. Binding of [125I]IGF I to monolayers of HSF also depends on NaCl under conditions where alpha IR-3 saturates the type I IGF receptor but does not affect IGF-BPs. These findings demonstrate that sodium chloride has a marked effect on the interaction of IGF-I with the type I IGF receptor in the plasma membrane and with BPs associated with the surface of intact HSFs. Since an effect is also evident in membranes prepared from intact tissues (human placenta), occurs at 4 C, and occurs with cells devoid of BPs, a mechanism involving receptor or BP translocation seems unlikely, at least as the sole explanation for these findings. Sodium ions (and other ions) may induce a conformational change in the receptor and BPs and cause decreased availability of both the IGF-I-binding site and the alpha IR-3 epitope on the receptor and the IGF-binding site on the BP. PMID- 2546752 TI - Iodine inhibits the proliferation of rat thyroid cells in culture. AB - We have explored the mechanisms whereby iodine inhibits thyroid growth using as models both the FRTL5 line of rat thyroid follicular cells that require TSH for growth and the M12 line of mutant cells that grow in the absence of TSH. Between 0.01-1.0 mM, NaI produced a dose-dependent inhibition of TSH stimulation of [3H]thymidine incorporation and replication in FRTL5 cells as well as spontaneous growth in M12 cells. Iodide also inhibited the cAMP-dependent growth of FRTL5 cells induced by forskolin and (Bu)2cAMP, as well as the cAMP-independent mitogenesis induced by insulin-like growth factor-I. The effect of iodide to inhibit both TSH- and insulin-like growth factor-I-stimulated growth in FRTL5 cells was abolished by concomitant culture with methimazole, and no iodide inhibition of growth was observed in L6 myoblasts and BRL 30E hepatocytes. Exposure of cells to iodide under conditions that resulted in inhibition of TSH stimulated growth did not significantly alter the ability of TSH to increase the intracellular cAMP concentration, nor did iodide alter two responses to TSH in FRTL5 cells that depend upon an increase in cAMP concentration: down-regulation of TSH receptor and cytoskeletal reorganization. We conclude that iodide exerts its inhibitory action on the growth of thyroid cells at multiple loci related to both the cAMP-dependent and cAMP-independent pathways of mitogenic regulation. PMID- 2546754 TI - Maturation and activation of hypothalamic-pituitary adrenal function in fetal sheep. AB - The progressive rise in the concentration of cortisol in the plasma of fetal sheep during late pregnancy arises from the sequential maturation of the fetal HPA axis. In addition, cortisol itself occupies a central role in accelerating this process through a series of feed-forward mechanisms. Before days 100-110 of pregnancy AVP appears to predominate over CRH as the major corticotropin releasing factor in the fetus. Pituitary responsiveness to CRH increases progressively after day 100, and precedes maturation of fetal adrenal responsiveness to endogenous or exogenous ACTH. Cortisol accelerates the increase in the ratio of adult-fetal corticotropes in the fetal pituitary. In addition, cortisol modulates the mechanism by which ACTH activates fetal adrenal function, possibly through an action at the level of the ACTH receptor. Cortisol appears also to mediate the rise in fetal plasma CBG concentrations during late pregnancy, and may thereby alter the efficacy of the negative feedback process. In women, cortisol acts on the placenta to promote rather than to inhibit CRH output. CRH from the placenta may reach significant concentrations in the fetal circulation and augment the drive to fetal ACTH release. It may also act in a paracrine fashion to promote placental POMC gene expression. The importance of placental CRF and ACTH in the sheep is not yet apparent. These feed-forward loops establish a series of positive cascades that ensure concurrent rises in plasma ACTH and cortisol in the fetal circulation during late pregnancy. We suggest that this sequence leads to, and is broken by, the process of birth. PMID- 2546753 TI - Sodium and potassium currents recorded during an action potential. AB - A simple method was used to measure directly sodium and potassium currents underlying the action potential in single nerve fibres of Xenopus laevis. A short rectangular stimulus under current-clamp conditions elicited an action potential which was digitally stored and later used as command when voltage-clamping the same fibre. The currents thus obtained nearly reproduced the original rectangular stimulus. Adding first 100 nM TTX and subsequently 100 nM TTX plus 10 mM TEA to the extracellular Ringer solution revealed the sodium and the potassium currents during an action potential. They were converted to permeabilities by use of the constant-field equation and are in good agreement with the curves which had been calculated from conventional voltage-clamp data. Thus experimentally determined currents and permeabilities are shown as they are changing during an action potential. PMID- 2546755 TI - Comparison of anamnestic history, alcohol intake and smoking, nutritional status, and liver dysfunction between thorotrast patients who developed primary liver cancer and those who did not. AB - In order to clarify the differences in past history, nutritional condition and, consumption of alcohol and tobacco, and liver dysfunction between the thorotrast patients who developed primary liver cancer and those who did not, 103 persons who had no primary liver cancer in January 1980 were studied. All subjects were military men who had undergone angiography with thorotrast between 1943 and 1946. Twenty persons developed hepatocellular carcinoma and 16 developed intrahepatic bile duct carcinoma by April 1987, whereas 67 are still alive without any cancer. There was no difference in age or period after thorotrast infusion between those two groups of patients in January 1980. A difference in history of hepatitis and/or jaundice and presence of hepatic dysfunction was found between the subjects who developed primary liver cancers and those who did not. These findings suggest that an anamnestic history of hepatitis and liver dysfunction are risks for development of thorotrast-induced liver cancer. On the basis of the above findings, early detection of liver dysfunction offers a possibility of early diagnosis of primary liver cancer. PMID- 2546757 TI - Superoxide generation by photomediated redox cycling of anthraquinones. AB - Anthraquinones (AQs) comprise one important class of secondary metabolites predominantly produced by fungi and higher plants but also produced by a variety of other organisms. Humans orally ingest AQs from environmental sources as well as through direct use as nonprescription laxatives, and some AQ derivatives are used as topically applied antipsoritic agents. Some AQs are mutagenic. We present evidence that aqueous solutions of several AQs in the presence of an appropriate reducing agent and dissolved oxygen generate superoxide when they are illuminated with broad-spectrum light. Redox cycling of AQs could be responsible for some aspects of their toxicity in biological systems. PMID- 2546756 TI - Plasma lipid peroxides among workers exposed to silica or asbestos dusts. AB - Plasma malondialdehyde (MDA) (an indicator of lipid peroxidation) was determined in random samples of workers (age range 25-60 years) exposed to silica dust (n = 31, mean exposure duration 21.3 +/- 8.3 years) or asbestos dust (n = 59, mean exposure duration 15.6 +/- 4.5 years) and in 52 age-matched healthy male controls. MDA levels of both exposed groups were significantly higher than that of the controls and were significantly higher among workers exposed to asbestos than among those exposed to silica dusts. Neither age nor smoking was related to MDA levels among both controls and exposed workers; among the latter group MDA was not significantly correlated with duration of exposure. Mean MDA levels of exposed workers with radiographic signs of lung fibrosis or pleural thickening and of those without such signs were not significantly different, except in the case of asbestos-exposed workers where it was significantly less than that of those without such signs. Differences between mean durations of dust exposure of workers with radiographic signs of lung fibrosis and those without such signs were statistically insignificant. The results suggest the possible involvement of lipid peroxidation on exposure to silica or asbestos dusts in humans and possible development of antioxidant mechanism(s) on prolonged dust exposure and support the opinion that development of pneumoconiosis depends on susceptibility factor(s). PMID- 2546758 TI - The human lysozyme gene. Sequence organization and chromosomal localization. AB - We have isolated two overlapping recombinant lambda-phage clones from a genomic lambda-EMBL3 library containing 25 kb of the human lysozyme gene region. Furthermore a full-lenght human lysozyme cDNA clone of 1.5 kb was isolated from a human placenta cDNA library. Nucleotide sequences of the entire structural gene and the cDNA clone were determined. The human lysozyme gene spans 5856 bp and its sequence organization with four exons and three introns is homologous to the chicken lysozyme gene and the human alpha-lactalbumin gene. Human and chicken lysozyme genes differ mainly in the size of their introns and 3' non-coding region. Four Alu repetitive elements were found in the human lysozyme gene, one in each intron and one on the fourth exon. Lysozyme transcripts of 1.6 kb and 0.6 kb in size were detected in human myeloid cell lines U-937, HL-60 and THP-1 and surprisingly in human hepatoma cell lines HepG2 and Hep3B. The lysozyme gene locus was assigned to human chromosome 12 by hybridization to a panel of DNAs from human-rodent somatic cell hybrids. PMID- 2546759 TI - Direct electron transfer of redox proteins at the bare glassy carbon electrode. AB - A simple system is presented for the microscale, direct voltammetry of redox proteins, typically 25 micrograms, in the absence of mediators and/or modifiers. The sample consists of a droplet of anaerobic solution laid onto an oversized disc of nitric-acid-pretreated glassy carbon as the working electrode. Very reproducible, Nernstian responses are obtained with horse heart cytochrome c. The midpoint potential Em (pH 7.0) is dependent on the ionic strength, ranging from $293 mV in 1 mM potassium phosphate to $266 mV in 0.1 M phosphate. At fixed buffer and cytochrome concentrations the magnitude of the voltammetric response is found to be independent of pH over six pH units around neutrality. It is suggested that the response of the present system is not complicated by pH dependent properties of the electrode surface around physiological pH and, therefore, that the use of this system is practical in biochemically oriented studies. Direct, quasi-reversible responses have also been obtained at pH 7.0 (5 mM phosphate) from Desulfovibrio vulgaris. Hildenborough strain, tetraheme cytochrome c3 (pI = 10.0 at 4 C; 3 X Em = -0.32 mV, Em = -0.26 V), and cytochrome c553 (pI = 9.3; Em = +60 mV), and from Megasphaera elsdenii rubredoxin (pI congruent to 3; Em = -353 mV). The latter protein absorbs onto the glassy carbon surface, thus forming a system with possible applications in the electrochemical study of ferredoxin-linked enzymes. PMID- 2546760 TI - The expression and purification of human rhinovirus protease 3C. AB - Human rhinovirus type 14 protease 3C was expressed as a soluble and active protein in Escherichia coli. The protease was purified by a cationic-exchange step followed by gel filtration on a TSK 3000 column. The final yield of purified protease was in the range 0.5-1.0 mg/l culture grown to A550 = 1.0. Sequence analysis revealed that greater than 90% of the N-terminal residues were methionine. The enzyme activity of the purified protease was measured by cleavage of a synthetic peptide representing a predicted Gln/Gly viral polyprotein cleavage site. A mutant protease (Cys146----Ser) was produced and purified in the same way. The yield of mutant protease 3C was approximately 150 micrograms/l from a culture grown to A550 = 1.0. This mutant protease 3C did not cleave the synthetic peptide substrate. PMID- 2546761 TI - The ATP/ADP-antiporter is involved in the uncoupling effect of fatty acids on mitochondria. AB - The ATP/ADP-antiporter inhibitors and the substrate ADP suppress the uncoupling effect induced by low (10-20 microM) concentrations of palmitate in mitochondria from skeletal muscle and liver. The inhibitors and ADP are found to (a) inhibit the palmitate-stimulated respiration in the controlled state and (b) increase the membrane potential lowered by palmitate. The degree of efficiency decreases in the order: carboxyatractylate (CAtr) greater than ADP greater than bongkrekic acid, atractylate. GDP is ineffective, Mg.ADP is of much smaller effect, whereas ATP is effective at much higher concentration than is ADP. Inhibitor concentrations, which maximally suppress the palmitate-stimulated respiration, correspond to those needed for arresting the state 3 respiration. The extent of the CAtr-sensitive stimulation of respiration by palmitate has been found to decrease with an increase in palmitate concentration. Stimulation of the controlled respiration by p-trifluoromethoxycarbonylcyanide phenylhydrozone (FCCP) and gramicidin D at any concentrations of these uncouplers is CAtr insensitive, whereas that caused by a low concentrations of 2,4-dinitrophenol and dodecyl sulfate is inhibited by CAtr. The above effect of palmitate develops immediately after addition of the fatty acid. It is resistant to EGTA as well as to inhibitors of phospholipase (nupercain) and of lipid peroxidation (ionol). Moreover, palmitate accelerates spontaneous release of the respiratory control, developing in rat liver mitochondria under certain conditions. This effect takes several minutes, being sensitive to EGTA, nupercain and ionol. Like the fast uncoupling, this slow effect is inhibited by ADP but CAtr and atractylate are stimulatory rather than inhibitory. In artificial planar phospholipid membrane, palmitate does not increase the membrane conductance, FCCP increases it strongly and dinitrophenol only slightly. In cytochrome oxidase proteoliposomes, FCCP, gramicidin and dinitrophenol (less effectively) lower, whereas palmitate enhances the cytochrome-oxidase-generated membrane potential. In this system, monensin substitutes for palmitate. It is concluded that the ATP/ADP antiporter is somehow involved in the uncoupling effect caused by low concentrations of palmitate and, partially, of dinitrophenol, whereas uncoupling produced by FCCP and gramicidin is due to their action on the phospholipid part of the mitochondrial membrane. A possible mechanism of this effect is discussed. PMID- 2546762 TI - The coupling between protonmotive force and the NAD(P)+ transhydrogenase in chromatophores from photosynthetic bacteria. AB - 1. The activity of NAD(P)+ transhydrogenase in chromatophores of Rhodobacter capsulatus relaxed from a high rate during illumination to a lower rate after darkening with a half-time of approximately 100 ms. 2. The dissipative ionic current flowing across the chromatophore membrane was increased in the presence of transhydrogenase substrates. This is attributed to proton current through the transhydrogenase enzyme. Subject to the assumption that transhydrogenase does not conduct in the absence of nucleotide substrates, the ratio of protons translocated across the membrane per hydride ion transferred was 0.4 +/- 0.5. Within the error and uncertainities in the calibration procedure, this ratio may be consistent with a stoichiometry of one but higher values seem unlikely. The ratio of hydride ion transferred in the transhydrogenase to electrons transferred through the cyclic electron transport system was approximately 0.2. 3. The Kappm values for the transhydrogenase substrates were determined for chromatophores in illuminated and darkened suspensions over a range of pH. These values are discussed in relation to the equivalent parameters reported for mitochondria transhydrogenase [Rydstrom, J. (1977) Biochim. Biophys. Acta 255, 9641-9646] and were used to calculate the concentrations of substrates which effectively saturate the enzyme. 4. At substrate concentrations which were in excess of 8 X Kappm the dependence of transhydrogenase rate on the value of the membrane potential (zero pH gradient) was determined at pH 6.3, 6.9, 7.6 and 9.0. The relation was similar at pH 6.9 and 7.6. At alkaline pH the apparent threshold in the relation became more prominent as it was shifted to slightly higher values of membrane potential. At acid pH a shift in the opposite direction diminished the apparent threshold and saturation at high membrane potential became more dominant. We use these data in an attempt to discriminate between two models of energy transduction: (a) the driving force exerted by the membrane potential is mediated by a pH gradient formed through the operation of a proton well in the transhydrogenase; (b) the membrane potential increases a rate constant for charge translocation through transhydrogenase by decreasing the effective height of the Eyring barrier for charge transfer across the membrane through the enzyme. The second model leads to a more simple description than the first of the pH dependence of transhydrogenase rate on membrane potential.4+ transhydrogenase activity in chromatopho PMID- 2546763 TI - Characterization of trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase of Saccharomyces cerevisiae. AB - The properties of yeast trehalose-6-phosphate synthase were reinvestigated in relation with the recent claim made by Panek et al. [Panek, A. C., de Araujo, P. S., Moura-Neto, V. and Panek, A. D. (1987) Curr. Genet. II, 459-465] that the enzyme would be stimulated by ATP and partially inactivated by cAMP-dependent protein kinase. Trehalose-6-phosphate synthase activity was measured by the sum of [14C]trehalose 6-phosphate and [14C]trehalose formed from UDP-[14C]glucose and glucose 6-phosphate. The activity measured in an extract of Saccharomyces cerevisiae was not affected by any treatment of the cells, such as incubation in the presence of glucose or of dinitrophenol, which are known to greatly increase the intracellular concentration of cAMP, nor by preincubation of the extract in the presence of ATP-Mg, cAMP and bovine heart cAMP-dependent protein kinase. The activity was also not significantly different in several mutants affected in the cAMP system. The kinetic properties of the partially purified enzyme were investigated; no effect of ATP could be detected but Pi acted as a potent noncompetitive inhibitor (Ki = 2 mM). The activity of trehalose-6-phosphate phosphatase was measured by the amount of [14C]trehalose formed from [14C]trehalose 6-phosphate. The enzyme could be separated from other phosphatases and appeared to be highly specific for trehalose 6-phosphate. It was Mg dependent and its kinetics for trehalose 6-phosphate was hyperbolic. Studies performed with intact cells, crude extracts or the purified enzyme did not reveal any cAMP dependent change in its activity. Remarkably, trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase copurified in the course of different chromatographic procedures, suggesting that they are part of a single bifunctional protein. A 50-fold purification of the two enzymes could be achieved with a yield of only 2% by chromatography on Mono S followed by gel filtration on Superose 6B. PMID- 2546764 TI - Tumor necrosis factor stimulates human neutrophils to release leukotriene B4 and platelet-activating factor. Induction of phospholipase A2 and acetyl-CoA:1-alkyl sn-glycero-3-phosphocholine O2-acetyltransferase activity and inhibition by antiproteinase. AB - Tumor necrosis factor stimulates polymorphonuclearneutrophils to synthesize leukotriene B4 and platelet-activating factor (PAF), but alpha 1-proteinase inhibitor and alpha 1-antichymotrypsin block this response. However, proteinases such as elastase and cathepsin G induce preferentially synthesis of PAF. An acetyltransferase required, together with phospholipase A2, in the remodeling pathway of PAF synthesis is activated in polymorphonuclearneutrophils stimulated by tumor necrosis factor and elastase. In contrast, 1-oleyl-2-acetylglycerol, a protein kinase C activator, promotes PAF formation by the de novo biosynthetic pathway without activating the acetyltransferase. Staurosporine, an inhibitor of protein kinase C, blocks PAF production apparently by inhibiting phospholipase A2. This suggests that diacylglycerols are involved in activating both pathway of PAF synthesis. PMID- 2546765 TI - A B-type PDGF receptor lacking most of the intracellular domain escapes degradation after ligand binding. AB - The characteristics of the human B-type platelet-derived-growth-factor (PDGF) receptor expressed in Chinese hamster ovary (CHO) cells, were compared with those of a mutant receptor lacking all but 19 amino acids of the intracellular domain. The transfected wild-type receptor was synthesized as a 160-kDa precursor that was processed to 190 kDa. Each CHO cell expressed 30,000-100,000 receptors which bound PDGF-BB with a Kd of about 0.5 nM. Analysis of PDGF-AB binding yielded non linear Scatchard plots; the major part of the binding sites had a Kd of 6 nM. PDGF-AA was not bound. The receptors expressed in CHO cells were down-regulated after binding of PDGF-BB, and mediated degradation of 125I-PDGF-BB with similar efficiency as PDGF-B-type receptors in human fibroblasts. The transfected receptor also transduced a mitogenic signal. The mutant receptor was synthesized as a 90-kDa precursor and was processed to 120 kDa with a slightly faster rate than the wild-type receptor. Cells expressing the mutant receptor generally had around 10(6) ligand-binding sites/cell, with a Kd for binding of PDGF-BB of 3 nM. The mutant receptor, which did not transduce a mitogenic response, mediated degradation of 125I-PDGF-BB, albeit less efficiently compared to the wild-type receptor. In contrast to the wild-type receptor, it was down-regulated only to a limited extent and not degraded in response to ligand binding. These findings indicate a role for the intracellular part of the receptor, not only in mitogenic signaling, but also in receptor internalization and intracellular routing. PMID- 2546766 TI - Purification of cytochrome-c oxidase retaining its pulsed form. AB - A new purification procedure for cytochrome-c oxidase from bovine heart mitochondria is described. The enzyme was purified by selective solubilization in Triton X-100 and subsequent hydroxyapatite and gel chromatography. The preparation was highly pure and active. The subunit composition and steady-state kinetics were found to be the same as those reported for other preparations. In contrast to most of the previously published protocols the method presented here resulted in a preparation which had a rapid intramolecular electron transfer from cytochrome a to cytochrome a3, i.e. it was found to have retained its pulsed state. This correlated with monoexponential cyanide-binding kinetics. The formation of resting kinetics and biphasic cyanide-binding kinetics was shown to be induced by a short incubation at pH 5.0. PMID- 2546768 TI - Transcription of the alpha A-globin gene of the duck. Development of a homologous in vitro system and identification of trans-acting factors. AB - A homologous in vitro transcription system was developed in which the alpha A globin gene of the duck was faithfully transcribed. Whole-cell extracts from duck erythrocytes were separated into fractions A, B, C and D by consecutive elution from phosphocellulose columns and were individually reconstituted in run-off transcription assays. Fractions A, C and D were required to achieve faithful initiation on the alpha A-globin gene. The latter fractions were mutually interchangeable with comparable fractions from HeLa cells. A fourth fraction, B, was not required but enhanced basal transcription when reconstituted with fractions A, C and D or a very low amount of HeLa whole-cell extract which by itself did not yield a detectable signal. Fraction B from duck erythrocytes was further purified by chromatography on DEAE-Sephadex and was shown to contain two trans-acting factors. One of these differentially acts on the alpha A-globin gene of the duck. The other component from duck erythrocytes surprisingly resembles the upstream stimulatory factor, previously isolated from HeLa cells. This latter protein binds to and trans-activates the adenovirus 2 major late promoter, but is not involved in the transcription of the alpha A-globin gene. PMID- 2546767 TI - Inhibition of NADPH oxidase by aminoacyl chloromethane protease inhibitors in phorbol-ester-stimulated human neutrophils: a reinvestigation. Are proteases really involved in the activation process? AB - Superoxide anion production by polymorphonuclear leukocytes stimulated with phorbol 12-myristate 13-acetate is known to be inhibited by a number of inhibitors and substrates of serine proteases, in particular by tosylphenylalanylchloromethane (TosPheCH2Cl) and to a lesser extent by tosyllysylchloromethane (TosLysCH2Cl). We have reinvestigated the characteristics of this inhibition, in view of the fact that other serine protease inhibitors with similar specificities, phenylmethanesulfonyl fluoride and leupeptin, were without effect. We found that the inhibition of phorbol-ester-induced superoxide production after cell preincubation with the chloromethanes followed saturation kinetics, with Kinact and kinact values of 100 microM and 31 min-1 for TosPheCH2Cl and 2 mM and 18 min-1 for TosLysCh2Cl. We also showed that the two compounds, which can inhibit protein kinase C in vitro, inhibited neither its activity in vivo, nor its translocation induced by phorbol myristate acetate. Furthermore the intracellular non-protein sulfhydryl group content was not affected by the treatment with the chloromethanes. Finally, addition of the inhibitors to stimulated cells also led to a time-dependent, concentration dependent inhibition of superoxide production. Altogether, our results suggest that the chloromethane target is neither a protease nor protein kinase C and is not involved in NADPH oxidase activation, but rather in maintenance of its activity. The possible identity of this protein is discussed. PMID- 2546769 TI - The primary structure of rat ribosomal protein L37a. AB - The amino acid sequence of rat ribosomal protein L37a was deduced from the sequence of nucleotides in recombinant cDNAs isolated in Yamagata and in Chicago and confirmed from the NH2-terminal amino acid sequence of the protein. Ribosomal protein L37a contains 91 amino acids (the NH2-terminal methionine is removed after translation of the mRNA) and has a molecular mass of 10143 Da. PMID- 2546770 TI - Evidence for at least four different inositol bisphosphatases in bovine brain. AB - Bovine brain supernatant contains at least four enzymes capable of hydrolysing inositol bisphosphates. These activities may be distinguished on the basis of their metal, salt and pH dependence, sensitivity to Li+ ions and thiol modification reagents, and on their molecular sizes. In addition to Li+-sensitive Ins(1,4)P2/Ins(1,3,4)P3 1-phosphatase [Gee et al. (1988) Biochem. J. 253, 777 782] which has an absolute requirement for Mg2+, two (Li+-insensitive and Mg2+ independent) phosphatases, capable of hydrolysing Ins(3,4)P2 and Ins(1,3)P2, respectively, have been identified. Both enzymes were inhibited by only moderate concentrations of salt, although for the former there was no obvious correlation between inhibitory potency and either the nature of the anion/cation or the ionic strength of the buffer. Ins(3,4)P2 phosphatase had a pH optimum of 7.6 and this activity could be resolved on gel-filtration columns into a two overlapping peaks of molecular mass 170 kDa and 450 kDa. Mg2+-independent Ins(1,3)P2 phosphatase had a pH optimum of 7.1 and displayed a single broad activity peak on gel filtration columns. However, if assays were performed in the presence of Mg2+, a second Ins(1,3)P2 phosphatase was revealed (35 kDa), which had a pH optimum of 8.8 Ins(1,4)P2/Ins(1,3,4)P3 1-phosphatase, Ins(3,4)P2 phosphatase, Mg2+ independent Ins(1,3)P2 phosphatase and inositol monophosphatase were all inhibited by 5,5'-dithiobis(2-nitrobenzoic acid) with IC50 values of 34 microM, 65 microM and 560 microM and 1100 microM, respectively. The metabolism of Ins(1,3,4)P3 by brain supernatant was also examined. Product specificity was shown to be entirely dependent on the buffer conditions employed. In Mg2+ containing buffers, Ins(1,3,4)P3 was hydrolysed predominantly to Ins(3,4)P2, consistent with hydrolysis by Ins(1,4)P2/Ins(1,3,4)P3 1-phosphatase. In the presence of EDTA, Ins(1,3,4)P3 was degraded exclusively by a 4-phosphatase enzyme generating Ins(1,3)P2. Under these conditions, high concentrations of Ins(3,4)P2 blocked the hydrolysis of Ins(1,3,4)P3. PMID- 2546771 TI - Interaction of non-classical detergents with the mitochondrial porin. A new purification procedure and characterization of the pore-forming unit. AB - The effect of different families of detergents on the solubilization and purification of the pore-forming protein (porin) of the mitochondrial outer membrane of bovine heart was investigated in detail. With Tritons, dimethylamine oxides and zwittergents, porin solubilization with respect to total mitochondrial membrane protein was more efficient with the more hydrophobic members of each series. With most detergents the protein eluted as protein-detergent micelles in the void volume of hydroxyapatite/celite columns. In contrast, the protein was bound to the column material and was eluted after the addition of salt to the elution buffer when the detergents octylglucoside, zwittergent Z-314 and lauryl(dimethyl)-amine oxide were used. The protein purified in the presence of the latter detergent had a higher pore-forming activity in lipid bilayer membranes compared to porin isolated in the presence of Triton X-100. The binding of porin to the hydroxyapatite/celite column was used to study the lipid content of the active pore-forming complex. The analysis revealed that the complex contained no phospholipid but rather five molecules of cholesterol/polypeptide chain. PMID- 2546772 TI - The pathway of the quinol/quinone transhydrogenation reaction in ubiquinol: cytochrome-c reductase of Neurospora mitochondria. AB - Ubiquinol: cytochrome-c reductase, isolated from Neurospora mitochondria as a protein/Triton X-100 preparation, and reconstituted into phospholipid membranes, catalyses the electron transfer from duroquinol to 2.3-dimethoxy-5-decyl-6-methyl benzoquinone (decQ) on a myxothiazol-insensitive, but antimycin-sensitive, ping pong pathway. Duroquinol reacts first to form the altered, reduced enzyme E'. This reaction is followed by dissociation of duroquinone making way for E' to bind decQ and convert it into decQH2. PMID- 2546773 TI - Factors relevant in the reaction of pyrroloquinoline quinone with amino acids. Analytical and mechanistic implications. AB - In order to reveal the stability of pyrroloquinoline quinone (PQQ) in complex samples, its reaction on incubation with amino acids was followed spectrophotometrically by monitoring oxygen consumption, and with a biological assay. For several alpha-amino acids, the formation of a yellow coloured compound (lambda max = 420 nm) was accompanied by oxygen uptake and disappearance of biological activity from the reaction mixture. The yellow product appeared to be an oxazole of PQQ, the exact structure depending on the amino acid used. Oxazole formation also occurred under anaerobic conditions with concomitant formation of PQQH2, suggesting that PQQ is able to oxidize the presumed oxazoline to the oxazole. Besides the condensation reaction, there is also a catalytic cycle in which an aldimine adduct of PQQ and the amino acid is converted into the aminophenol form of the cofactor and an aldehyde resulting from oxidative decarboxylation of the amino acid. Addition of NH4+ salts, as well as that of certain divalent cations, greatly stimulated both the cyclic and the linear reaction. With basic amino acids, oxazole formation scarcely occurred. However, as oxygen consumption was observed (provided that certain divalent cations were present), conversion of these compounds took place. A reaction scheme is proposed accounting for the products formed and the effects observed. Since NH4+ ions activate several quinoproteins (PQQ-containing enzymes) and divalent cations (Ca2+, Fe2+, and Cu2+) are additional (co)factors in certain metallo quinoproteins, the effects of metal ions observed here could be related to the mechanistic features of these enzymes. Although all oxazoles were converted to PQQ by acid hydrolysis, PQQ was not detected when hydrolysis was carried out in the presence of tryptophan, a compound which appeared to have a deleterious effect on the cofactor under this condition. The results here described explain why analysis methods for free PQQ in complex samples fail in certain cases, or are not quantitative. PMID- 2546774 TI - Testing of organisms for susceptibility to triazoles: is it justified? PMID- 2546776 TI - Left ventricular dimensions during isometric exercise in aortic valve incompetence assessed by M-mode echocardiography and gated equilibrium radionuclide angiography. AB - We compared M-mode echocardiographic and gated equilibrium radionuclide angiography assessment of the left ventricular (LV) dimensions at rest and during isometric exercise in 18 patients with chronic aortic valve incompetence. The two methods showed a satisfactory correlation when comparing LV size at rest and during exercise (LV end-diastolic dimension in echocardiography vs LV end diastolic volume in radionuclide angiography, r = 0.80, P less than 0.01 at rest and r = 0.81, P less than 0.01 at rest and r = 0.75; P less than 0.01 during exercise), but fractional shortening in echocardiography and ejection fraction in radionuclide angiography did not correlate (r = 0.27, not significant (NS) at rest and r = 0.34, NS during exercise). Thus echocardiography and radionuclide angiography describe LV dimensions at rest and during handgrip exercise in a similar fashion, documenting the concordance of these noninvasive methods to describe LV size in aortic incompetence at rest and during exercise. PMID- 2546777 TI - Intensive chemotherapy in SCLC. PMID- 2546775 TI - New antifungal agents. AB - For more than two decades, amphotericin B has been the single broad-spectrum agent for the treatment of systemic mycoses. Amphotericin B is not always effective, must be given parenterally, and is associated with a host of adverse reactions. Despite amphotericin B toxicity, until recently the systemic mycoses did not rate enough attention to prompt a search for new alternatives. However, three recent events have overcome this inertia: the gradually increasing use of potent immunosuppressive agents and broad-spectrum antibacterial drugs; the discovery of the relatively nontoxic azole classes of antifungal drugs in the 1980s and the rapid emergence of AIDS, with its severe accompanying opportunistic fungal infections. In just ten years we have seen the emergence of second generation imidazole and third-generation triazole antifungal drugs and, most recently, entirely new classes of agents. It is remarkable that so many alternatives are becoming available just at the time when new antifungal drugs have become a major need. This discussion will concentrate on the new antifungal drugs of the past ten years, with the exception of developments in the polyenes and flucytosine, which are covered elsewhere. PMID- 2546778 TI - A randomized clinical trial comparing systemic radiotherapy versus chemotherapy versus local radiotherapy in small cell lung cancer. AB - Between 1982 and 1987 a prospectively randomized trial of sequential hemibody irradiation (SHBI) (A), a non-cross-resistant chemotherapy drug combination (B) and local and/or locoregional radiotherapy (C) in small cell lung cancer (SCLC) was conducted. Previously untreated patients with extensive SCLC were randomized into three arms: A = 31 patients, B = 37, C = 31. In the chemotherapy combination, the following were used: etoposide, doxorubicin, methotrexate (VAM) and procarbacine, vincristine, cyclophosphamide, lomustine (POCC) and prophylactic cranial irradiation (30 Gy). The results show that the median survival was significantly (P less than 0.01) better in chemotherapy (44 weeks) compared with 17 and 20 weeks in arms A and C, respectively. One year and 2 year survival rates were better for the chemotherapy arm. No differences were found between groups A and C. In comparing the total hospitalization time expressed as a percentage of overall survival, an advantage for group B was shown. In conclusion, high dose SHBI cannot be recommended as a standard therapy for extensive SCLC. PMID- 2546779 TI - Reticulum cell sarcomas of SJL mice have rearranged immunoglobulin heavy and light chain genes. AB - The immunoglobulin (Ig) heavy (H) and light (L) chain gene rearrangements of the high incidence SJL lymphomas (reticulum cell sarcoma, RCS) have been analyzed. Both primary and transplanted RCS show rearrangements of H and kappa L chains, demonstrating that these tumors are of B cell origin. These data are consistent with previous results indicating that these tumors are a mouse model for follicular lymphoma. A long-term transplanted line and the in vitro line derived from it, cRCS-X, have a single rearranged JH-C gamma 2a fragment and one rearranged C alpha gene fragment which does not hybridize with a probe for the JH gene segments. These cell lines also have two rearranged J kappa-C kappa fragments. Primary tumors and early passages are more heterogeneous with respect to Ig gene rearrangements, possibly because more than one B cell clone is present. Although no synthesis of IgG2a, or of any Ig, could be detected by the in vitro cRCS-X cells, these cells contain abundant poly(A)+ RNA that hybridize with gamma 2a and kappa probes as well as lesser amounts of alpha and epsilon RNA. None of these H chain RNA hybridized with probes for the JH gene segments. The epsilon and alpha RNA are the same size as transcripts of germ-line CH genes which have been identified in other systems. However, the gamma 2a RNA are smaller than previously described germ-line C gamma 2a RNA and appear to be transcribed from aberrantly rearranged JH-C gamma 2a genes. PMID- 2546780 TI - Increased cyclic AMP levels block interleukin 2-induced protein kinase C substrate phosphorylation but not the mitogenic response. AB - Protein kinase C (PKC) has been implicated in the signaling of a number of cellular responses including activation of T cells. In the present report we have evaluated the effect of increased cAMP levels on PKC activation after stimulation of two distinct receptor systems on normal human T cells. PKC substrate phosphorylation can be induced via either the CD3 complex or, to a limited extent, the high affinity interleukin 2 (IL 2) receptor. Substrate phosphorylation via both pathways is shown to be blocked by increased intracellular levels of cAMP. In accordance with previous reports, the CD3 dependent autocrine proliferative response could also be blocked by a cAMP dependent mechanism. Since direct activation of PKC with a phorbol ester reversed this inhibition, a causal relationship between cAMP-dependent PKC blockage and inhibition of the CD3 response is suggested. In contrast, however, initiation of IL 2-induced proliferation was essentially unaltered by cAMP and could progress in the apparent absence of PKC activity. Thus, this study indicates that IL 2 induced proliferation can under such conditions be completely uncoupled from IL 2 induced PKC activation in normal T cells. PMID- 2546781 TI - E-series prostaglandins are potent growth inhibitors for some B lymphomas. AB - The ability of prostaglandins (PG) to inhibit the growth of B cell lymphomas was investigated. Macrophage-secreted PGE2 was previously shown to promote unresponsiveness to antigen in normal B lymphocytes. This observation suggested that B lymphomas might also be regulated by prostanoids. Five non-PG-secreting Ly 1+ B lymphomas (CH12, CH31, CH33, NBL and WEHI-231) were incubated for 24-72 h with PGE2, PGE1 or PGF2 alpha. The level of lymphoma growth at the end of culture was determined using a colorimetric assay which detects only viable cells. A marked heterogeneity was observed with respect to the sensitivity of these lymphomas to PGE2 and PGE1. CH31 was very sensitive, being growth inhibited by as little as 10(-8) M PGE. In contrast, CH12, a more mature lymphoma, was highly resistant, whereas CH33, NBL and WEHI-231 were of intermediate resistance. All five lymphomas demonstrated little or no growth inhibition when cultured with PGF2 alpha. Moreover, unlike PGE2, PGF2 alpha failed to elevate intracellular cAMP levels. It was previously shown that CH31, CH33 and WEHI-231 could be growth inhibited by anti-immunoglobulin antibodies which cross-link surface immunoglobulin. Interestingly, these three lymphomas were rendered more sensitive to this treatment if PGE2 was present. For example, 10(-8) M PGE2 alone had little effect on CH33, but significantly augmented growth inhibition induced by suboptimal quantities of anti-immunoglobulin antibody. Cholera toxin, another agent which was found to rapidly elevate intracellular cAMP levels, also synergized with suboptimal doses of anti-immunoglobulin to induce growth inhibition. Overall these data suggest that, in vivo, macrophage-secreted prostanoids may slow the growth of some lymphomas and that anti-immunoglobulin or anti-idiotype treatment may be more effective in the presence of agents which elevate cAMP such as E-series PG. PMID- 2546782 TI - Human endothelial cells inhibit platelet aggregation by separately stimulating platelet cyclic AMP and cyclic GMP. AB - Thrombin-induced platelet aggregation was monitored in a mixture of washed human platelets and cultured human endothelial cells on microcarrier beads. Endothelial cells completely inhibited platelet aggregation and enhanced the content of both cyclic AMP and cyclic GMP in the platelets. Inhibition of endothelial prostacyclin synthesis with indomethacin abolished the cyclic AMP increase in the platelets, whereas the cyclic GMP increase was unimpaired. A significant component of the endothelial anti-aggregatory effect persisted in the presence of indomethacin. This anti-aggregatory component and the elevation of cyclic GMP were blocked by the inhibitors of endothelium-derived relaxing factor (EDRF), gossypol, haemoglobin and methylene blue. These inhibitors did not affect the endothelium-induced increases in cyclic AMP or the cyclic AMP-mediated anti aggregatory effects. Exogenous prostacyclin stimulated only platelet cyclic AMP, whereas sodium nitroprusside selectively enhanced cyclic GMP. These data suggest that the endothelium can inhibit platelet aggregation by two completely separate mechanisms, one mediated by prostacyclin and cyclic AMP, and the other by EDRF and cyclic GMP. PMID- 2546783 TI - Autoradiographic localization of changes in pulmonary beta-adrenoceptors in an animal model of atopy. AB - Vaccination of guinea pigs with Haemophilus influenzae leads to an impairment of beta-adrenoceptor function in lung. We have used an autoradiographic technique to study the distribution of changes in lung beta-adrenoceptor density. H. influenzae induced a decrease in beta-adrenoceptors in peripheral lung membranes of 22 +/- 5% (mean +/- S.E.M., n = 7), while the affinity of binding was unaffected. Tracheal beta-adrenoceptor binding was not influenced by H. influenzae. Autoradiography revealed a 27% reduction in beta-adrenergic binding sites on alveolar septa. Bronchial epithelial beta-adrenoceptors were decreased for 36%, and vascular smooth muscle and endothelial beta-adrenoceptors were also reduced. beta-Adrenoceptors on airway smooth muscle were unaffected. H. influenzae affected both the beta 1- and beta 2-subtypes of receptors. It is concluded that in this animal model of atopy beta-adrenoceptors may be decreased on several different cell types within the lungs, which may influence overall airway and vascular reactivity. PMID- 2546785 TI - Chlormethiazole: neurochemical actions at the gamma-aminobutyric acid receptor complex. AB - Chlormethiazole has been extensively employed as a sedative/hypnotic and anticonvulsant for more than 25 years. While pharmacological and electrophysiological studies have implicated the GABAA receptor complex in these actions, neurochemical findings have not been consistent with this conclusion. We now present evidence that pharmacologically relevant concentrations of chlormethiazole perturb the GABAA receptor complex. Chlormethiazole was found to increase 36Cl- uptake into rat cortical synaptoneurosomes in a concentration dependent (EC50 = 48 +/- 3 microM; Emax = 8.9 +/- 0.8 nmol Cl-/mg protein per 5 s), picrotoxin-sensitive fashion. Chlormethiazole was also found to inhibit the binding of the 'cage' convulsant [35S]t-butylbicyclophosphorothionate to rat cortical membranes (IC50 = 58.6 +/- 0.6 microM) through an increase in the apparent KD of this radioligand. Moreover, at these concentrations chlormethiazole did not affect pentobarbital-enhanced [3H]flunitrazepam binding, but inhibited [3H]flunitrazepam binding with a low potency (IC50 = 1.6 +/- 0.2 mM). These findings provide neurochemical evidence that pharmacologically relevant concentrations of chlormethiazole can perturb the GABAA receptor complex, and suggest that this compound acts at a distinct locus from other sedative/hypnotics such as barbiturates, benzodiazepines and GABAmimetics. PMID- 2546784 TI - Comparison of beta 1- and beta 2-adrenoceptor effects on gastric emptying of a fat meal in mice. AB - The effects of intraperitoneal administration of isoproterenol (a mixed beta 1- and beta 2-agonist), dobutamine (a selective beta 1-agonist) and clenbuterol (a selective beta 2-agonist) on gastric emptying were investigated in mice killed 30 min after gavage with a 51Cr radiolabeled milk meal. Administered 30 min prior to the meal, isoproterenol (2 mg/kg) and dobutamine (1 and 2 mg/kg) accelerated gastric emptying and clenbuterol (0.05 and 0.1 mg/kg) slowed it. The stimulation of gastric emptying by isoproterenol (2 mg/kg) and dobutamine (2 mg/kg) was blocked by the previous administration of either mixed beta 1- and beta 2 antagonist, propranolol (1 mg/kg), or a specific beta 1-antagonist, acebutolol (1 mg/kg). Only propranolol (1 mg/kg) antagonized the decrease of gastric emptying induced by clenbuterol. A high dose of propranolol (10 mg/kg), injected alone, reduced gastric emptying. It is concluded that the increase in gastric emptying caused by beta-adrenergic drugs is mediated through beta 1-adrenoceptors, while beta 2-receptors are involved in the inhibition of this function. The lack of effects of propranolol at doses able to block the action of beta 1- and beta 2 adrenoceptor agonists suggests that beta-adrenoceptors are not physiologically involved in the control of gastric emptying. PMID- 2546786 TI - Presynaptic alpha 2-adrenoceptor modulation of 5-hydroxytryptamine and noradrenaline release from vascular adrenergic nerves. AB - Modulation of the stimulation-evoked release of 5-hydroxytryptamine (5-HT) and noradrenaline (NA) by presynaptic alpha 2-adrenoceptors was characterized in the perfused mesenteric vascular bed of the rat. The vasoconstrictor response to periarterial nerve stimulation (PNS; 8 Hz), previously abolished in the presence of 30 nM prazosin, was restored after 15 min treatment with 10 microM 5-HT, without a significant effect on the pressor response to 1 nmol of infused NA, which was previously abolished with prazosin. The restored pressor response to PNS was abolished by 100 nM tetrodotoxin and 100 nM ketanserin. Clonidine (1-10 microM) in the presence of prazosin induced a dose-dependent potentiation of the restored pressor response to PNS after 5-HT treatment while BHT 920 (10 nM-1 microM) and 100 nM clonidine inhibited the restored response. In the presence of 100 nM phentolamine, the restored pressor response to PNS was not altered by clonidine, but was inhibited by BHT 920. The PNS (8 Hz)-evoked tritium release in a preparation labeled with [3H]5-HT was facilitated by clonidine (100 nM-10 microM) while BHT 920 (10 nM-1 microM) and cocaine (1-10 microM) reduced the release. Yohimbine (1 microM) antagonized the effects of clonidine and cocaine but not of BHT 920 on the PNS-evoked tritium release. In the preparation labeled with [3H]NA, clonidine did not alter the PNS-evoked tritium release while BHT 920 inhibited it and cocaine facilitated it. Yohimbine did not antagonize the effect of BHT 920.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546787 TI - Electrophysiological characteristics of neurons in neocortical explant cultures. AB - We examined the electrophysiological and morphological properties of neocortical neurons maintained in explant cultures prepared from the parietal cortex of newborn Sprague-Dawley rats. After 3-6 weeks in vitro, cultures showed regional differences in cellular density reminiscent of cortical layering, and an abundance of axonal processes. Pyramidal-shaped neurons with spinous dendrites were the dominant elements revealed by Lucifer yellow injections. Intracellular recordings revealed that many electrophysiological properties of neurons in the explants resembled those of neocortical neurons in vivo and in slice preparations. In response to depolarizing current injection, neurons in the explants showed the same three patterns of repetitive firing described in neocortical slices, as well as a similar array of responses. Spontaneous synaptic potentials were recorded from all neurons and complex PSPs were evoked in response to focal extracellular stimulation. GABAa receptors mediated a significant component of the evoked responses. Fifteen of sixty neurons generated action potentials that arose spontaneously from resting potentials. Neurons in many slices generated large, prolonged depolarizing potentials that reflected coordinated synaptic activity within the explants. These results underscore the usefulness of the neocortical explant as a valuable model for studying aspects of the behavior of circuits of cortical neurons. PMID- 2546788 TI - Effect of interleukin-1 on adrenocortical activity in intact and hypothalamic deafferentated male rats. AB - The present study was designed to elucidate the site of action of interleukin 1 (IL-1) modulation of the hypothalamic-hypophyseal-adrenal (HHA) axis. An intraperitoneal injection of recombinant human IL-1 beta (160 U/rat) significantly elevated serum levels of ACTH and corticosterone (CS). In rats with complete mediobasal hypothalamic deafferentation, the HHA response to IL-1 was inhibited. An intracerebroventricular injection of rIL-1 (2 U/rat) caused a marked increase in serum ACTH and CS. These results suggest that IL-1 activates the HHA axis by a direct effect upon the brain, and that intact neural connections between the mediobasal hypothalamus and extrahypothalamic brain regions are essential for IL-1-induced HHA responses. PMID- 2546790 TI - Effect of repeated versus single electroconvulsive seizures on adrenergic mediated phosphatidylinositol hydrolysis in rat neocortex. AB - Adrenergic-stimulated phosphatidylinositol (PI) hydrolysis was measured in cortical slices obtained from adult rats following electroconvulsive seizures (ECS). One group of animals received ECS daily for 15 days and a second group received a single ECS. Rats were then sacrificed at intervals of 15 min, 60 min, 4 h, and 24 h after the last ECS. Inositol 1-monophosphate (IP1) accumulation was transiently reduced (20%, P less than 0.01), at 15 min, in repeatedly shocked versus sham-shocked control animals. No changes were observed at later intervals nor at any time in rats submitted to a single ECS. These findings suggest that repeated but not single convulsive seizures transiently desensitize adrenergic mediated PI metabolism. Although repeated ECS significantly increased the density (Bmax) of alpha 1 recognition sites in cortical slices at 15 min, 4 h, and 24 h, this upregulation was not coupled to a functional change in PI hydrolysis. PMID- 2546789 TI - Neural regulation of mRNA for the alpha-subunit of acetylcholine receptors: role of neuromuscular transmission. AB - Levels of mRNA for acetylcholine receptor (AChR) subunits are relatively low in innervated skeletal muscles. Following denervation they rise rapidly, leading to increased AChR synthesis. The mechanism by which motor nerves normally regulate these mRNA levels is not yet known. In order to determine the possible role of synaptic transmission in this process, we have compared the effect of blockade of cholinergic ACh transmission with that of surgical denervation. Blockade of quantal ACh transmission was produced by injection of type A botulinum toxin into the soleus muscles of rats. We measured mRNA for the alpha-subunit of the AChR (alpha-AChR mRNA) in RNA extracts of botulinum-treated, denervated, and normal control muscles by hybridization with a highly specific cDNA probe. Our findings show that treatment with botulinum toxin resulted in an increase in alpha-AChR mRNA which was similar to the effect of surgical denervation, although slower in its time course. Since botulinum toxin specifically inhibits quantal ACh release, these results support the concept that cholinergic synaptic transmission plays a key role in mediating the neural control of the alpha-AChR message. The difference between the effects of denervation and botulinum-treatment may be explained by the fact that botulinum toxin does not block the spontaneous non quantal component of ACh transmission, which has previously been shown to have a partial influence in regulating certain properties of muscles. The present results suggest that synaptic transmission has an important influence in regulating gene expression in the target cell. PMID- 2546791 TI - Physiological properties of sensory neurons of the interstitial nucleus in the spinal trigeminal tract. AB - Neurons were recorded from the interstitial nucleus of the spinal trigeminal tract. They were all nociceptive specific and some projected to the parabrachial area. These data suggest that this nucleus can be regarded as a rostral extension of lamina I of trigeminal subnucleus caudalis. PMID- 2546792 TI - Tritrichomonas foetus: purification and characterization of hydrogenosomal ATP:AMP phosphotransferase (adenylate kinase). AB - The hydrogenosomal enzyme ATP:AMP phosphotransferase (adenylate kinase) (EC 2.7.4.3) was purified to apparent homogeneity from the bovine parasite Tritrichomonas foetus. A fraction enriched for hydrogenosomes was obtained from cell homogenates which had been subjected to differential and isopycnic centrifugation. Adenylate kinase was solubilized in 50 mM Tris-HCl, pH 7.3, containing 0.8% Triton X-100, and purified by sequential Affi-Gel blue affinity chromatography and high-performance liquid chromatography gel filtration. The purified enzyme, a monomer of Mr 29,000, exhibited Km values of 100, 195, and 83 microM for ADP, ATP, and AMP, respectively. Substituting other mono-, di-, and trinucleotides for AMP, ADP, and ATP gave less than half the maximal activity. Full enzyme activity requires Mg2+, but Mn2+ and Co2+ yield half maximal activity. The enzyme has a broad optimal pH range between pH 6 and 9. The enzyme was competitively inhibited by P1,P5-di(adenosine-5')pentaphosphate, a specific adenylate kinase inhibitor: the Ki was 150 nM. The enzyme was also inhibited with 5,5'-dithiobis(2-nitrobenzoic acid), and this inhibition could be reversed by the addition of 2 mM dithiothreitol. T. foetus adenylate kinase has similar catalytic and physical properties to that of the biologically closely related human parasite Trichomonas vaginalis. PMID- 2546794 TI - The primary structure of Pseudomonas cytochrome c peroxidase. AB - The primary structure of Pseudomonas cytochrome c peroxidase is presented. The intact protein was fragmented with cyanogen bromide into five fragments; partial cleavage was observed at a Met-His bond of the protein. The primary structure was established partly by automatic Edman degradations, partly by manual sequencing of peptides obtained with trypsin, thermolysin, chymotrypsin, pepsin, subtilisin and Staphylococcus aureus V8 endopeptidase. The order of the cyanogen bromide fragments was further confirmed by overlapping peptides obtained by specific cleavage of the whole protein. Pseudomonas cytochrome c peroxidase consists of 302 amino acid residues giving a calculated Mr of 33690. PMID- 2546793 TI - Leishmania donovani: isolation and characterization of sodium stibogluconate (Pentostam)-resistant cell lines. AB - Leishmania donovani promastigotes were generated by virtue of their resistance to incrementing concentrations of sodium stibogluconate (Pentostam) under completely defined growth conditions. The PENT0400 and PENT03200 cell lines were isolated after prolonged exposure to 0.4 mg/ml and 3.2 mg/ml Pentostam (Sb concentration), respectively. Whereas the effective concentration of Pentostam which inhibited the growth of wild type cells by 50% (EC50 value) was 0.1-0.15 mg/ml, the EC50 values for the PENT0400 and PENT03200 cells were approximately 1 and 4 mg/ml, respectively. The decreased sensitivities of both PENT0400 and PENT03200 cells to Pentostam were maintained after 6 months of continuous culture in the absence of selective pressure, indicating that the Pentostam resistance in the mutant organisms was a stable genetic trait. Interestingly, wild type and PENT03200 cells were equally sensitive to growth inhibition and cytotoxicity caused by SbCl5 and SbCl3, as well as to a variety of other cations such as Cd, Zn, and As. Wild type and PENT03200 cells also displayed equivalent growth sensitivities to a spectrum of other antiprotozoal agents, including antimony potassium tartrate, melarsoprol, pyrimethamine, pentamidine, formycin B, and difluoromethylornithine. These results illustrate a potentially useful model system to study Pentostam resistance in Leishmania and suggest that Pentostam resistance in vitro may be independent of antimony toxicity. PMID- 2546795 TI - Extracellular Na+ removal enhances granule secretion in platelets--evidence that Na+/H+ exchange is inhibitory to secretion induced by some agonists. AB - The effect of extracellular Na+ [( Na+]e) removal on agonist-induced granule secretion in platelets in relation to [pH]i and [Ca2+]i changes was investigated. Substitution of [Na+]e with choline+ of K+ resulted in a significant enhancement of 5HT secretion induced by thrombin, collagen, U46619 and the protein kinase C activators, PMA and diC8. Increases in [Ca2+]i induced by thrombin and U46619 were slightly inhibited or unaffected in these buffers, but [pH]i increases induced by thrombin, U46619, PMA and diC8 were abolished and a drop in [pH]i (0.05 0.1 units below resting) was observed. Although preincubation with potassium acetate produced a big drop in [pH]i and greatly increased secretion with all the agonists, particularly in the absence of [Na+]e, clear evidence that [pH]i rises due to Na+/H+ exchange are inhibitory to secretion was obtained only with thrombin. Thus, (i) NH4Cl, which restored the increase in [pH]i in the absence of [Na+]e reduced the potentiated secretory response to thrombin, (ii) no increase in thrombin-induced secretion was observed when Na+ was replaced with Li+, which allowed a normal increase in [pH]i and (iii) ethyl isopropyl amiloride (EIPA) abolished the [pH]i rise and potentiated thrombin-induced secretion. With collagen and U46619, the results suggest that removal of [Na+]e per se rather than inhibition of Na+/H+ exchange results in enhanced secretion. It is concluded that [Na+]e per se and [pH]i elevations via Na+/H+ exchange both have important inhibitory roles in the control of platelet granule secretion. PMID- 2546796 TI - Potentiation by adrenaline of thrombin-induced elevation of pHi is not essential for synergistic activation of human platelets. AB - Gel-filtrated human platelets were stimulated with thrombin in the absence and presence of adrenaline. Adrenaline markedly enhanced the thrombin-induced increase in cytoplasmic pH (pHi) in BCECF-loaded platelets. This rise in pHi was strongly inhibited by the Na+/H+ exchange blocker EIPA. The potentiation by adrenaline of thrombin-induced PLC activation measured as [32P]PA formation and final platelet responses was, however, not blocked by EIPA, even at low concentrations of thrombin. These results indicate that the enhancement by adrenaline of thrombin-induced cytoplasmic alkalinization may be a secondary effect which is not essential for the potentiation by adrenaline of platelet activation by thrombin. PMID- 2546797 TI - Different selectivities of oxidants during oxidation of methionine residues in the alpha-1-proteinase inhibitor. AB - Oxidation of the reactive site methionine (Met) in alpha-1-proteinase inhibitor (alpha-1-PI) to methionine sulfoxide (Met(O] is known to cause depletion of its elastase inhibitory activity. To estimate the selectivity of different oxidants in converting Met to Met(O) in alpha-1-PI, we measured the molar ratio Met(O)/alpha-1-PI at total inactivation. This ratio was determined to be 1.2 for both the myeloperoxidase/H2O2/chloride system and the related compound NH2Cl. With taurine monochloramine, another myeloperoxidase-related oxidant, 1.05 mol Met(O) were generated per mol alpha-1-PI during inactivation. These oxidants attack preferentially one Met residue in alpha-1-PI, which is identical with Met 358, as concluded from the parallelism of loss of elastase inhibitory activity and oxidation of Met. A similar high specificity for Met oxidation was determined for the xanthine oxidase-derived oxidants. In contrast, the ratio found for ozone and m-chloroperoxybenzoic acid was 6.0 and 5.0, respectively, indicating oxidation of additional Met residues besides the relative site Met in alpha-1-PI, i.e. unselective action of these oxidants. Further studies were performed on the efficiency of oxidants for total depletion of the elastase inhibitory capacity of alpha-1-PI. Ozone and m-chloroperoxybenzoic acid were 10-fold less effective and the superoxide anion/hydroxyl radicals were 30-50-fold less effective to inactivate the elastase inhibitory activity as compared to the myeloperoxidase derived oxidants. The myeloperoxidase-related oxidants are discussed as important regulators of alpha-1-PI activity in vivo. PMID- 2546798 TI - Heterogeneous distribution of nuclear triiodothyronine receptors in liver and preadipose cells as evidenced by their reactivity to antibodies against different protein products of erb A oncogenes. AB - Polyclonal antibodies raised in rabbits against bacterially produced peptides in the C-terminal region of v-erb A or human c-erb A alpha oncogenes recognize the nuclear triiodothyronine (T3) receptors in the T3-sensitive Ob 17 mouse preadipocyte cell line and not in mouse or rat liver. The results confirm the existence of different T3 receptors in different tissues. The results also suggest a heterogeneous receptor distribution within the preadipose cell line, with a predominance of c-erb A alpha-type species. Antibodies raised against domain 149 227, but not against domain 245-325, impair T3 binding, suggesting a role for this domain in ligand binding. PMID- 2546800 TI - ATP-dependent peptide release from mitochondria of reticulocytes. AB - ATP-dependent release of TCA-precipitable peptides from mitochondria-containing stroma (MCS) is described. The process is independent of ubiquitin, but is sensitive to hemin and to heat treatment. Neither chloramphenicol nor EGTA inhibit. 50% of the activity is dependent on charged tRNA. The peptides released from MCS possess a molecular mass of about 1-5 kDa and are degraded to TCA soluble compounds by a cytosolic protease system (fraction II) without ubiquitin. PMID- 2546799 TI - Electron transfer reactions in photosystem I following vitamin K1 depletion by ultraviolet irradiation. AB - Photosystem I preparations were irradiated with UV to destroy vitamin K1 in situ. The depletion of vitamin K1 resulted in inactivation of NADP+ photoreduction and introduction of a approximately 220 ms component in the flash generated P700+ rereduction at room temperature. The photoreduction of the terminal FeS centers FA and FB in control and vitamin K1-depleted preparations at 7 K were comparable. The data confirm that vitamin K1 is functionally implicated in primary electron transfer reactions in PS I at physiological temperature, and that the anomalous results at cryogenic temperature may be explicable in terms of a by-pass of the vitamin K1 acceptor site or heterogeneity introduced into the photosystem by quinone removal. PMID- 2546801 TI - Collagenase gene expression in fibroblasts is regulated by a three-dimensional contact with collagen. AB - Collagenase activity in fibroblasts is regulated by cytokines and the interaction with the extracellular matrix. In this study we demonstrate that fibroblasts cultured within a three-dimensional collagen gel show a strong induction of collagenase gene expression. In addition to increased de novo synthesis most of the secreted enzyme was found to be activated leading to a high collagenolytic activity and complete degradation of collagen matrices after removal of fetal calf serum. Collagen I gene expression was found to be reduced under these conditions. These data suggest a specific modulation of cellular metabolism in response to contact with a three-dimensional collagenous matrix resulting in the divergent regulation of collagen and collagenase. PMID- 2546802 TI - Receptor-independent metabolism of platelet-activating factor by myelogenous cells. AB - Human neutrophils incorporate and metabolize platelet-activating factor (PAF). We dissociated these events from PAF binding to its receptors. Cells were pretreated with either pronase, a PAF antagonist (L652731), or excess PAF. This reduced PAF receptor numbers by 70 to almost 100% but had no comparable effect upon the neutrophil's ability to metabolize PAF. Furthermore, HL-60 cells efficiently metabolized, but did not specifically bind, PAF. Thus, PAF receptor availability did not correlate with PAF metabolic capacity and we conclude that myelogenous tissues can process this bioactive ligand by a receptor-independent pathway. PMID- 2546803 TI - Transducin GTPase provides for rapid quenching of the cGMP cascade in rod outer segments. AB - The role of transducin GTPase in rapid cGMP phosphodiesterase quenching was studied by simultaneous registration of GTP hydrolysis and phosphodiesterase activity in the same rod outer segments (ROS) preparation. The results thus obtained allow the conclusion that: (i) phosphodiesterase quenching coincides with transducin-bound GTP hydrolysis independently of ROS concentration; (ii) an increase in the ROS concentration results in the acceleration of cascade quenching due to the existence of a GTPase accelerating mechanism in ROS; (iii) approximation to physiological conditions (protein concentration, temperature) provides a transducin GTPase rate equal to 1-2 turnovers per second i.e., sufficiently high for satisfying the real rate of photoresponse reversion in dark adapted rods. PMID- 2546804 TI - Protein kinase C is crucial in signal transduction during IFN-gamma induction in endothelial cells. AB - We have demonstrated that IFN-gamma, a potent peptide mediator in inflammatory responses, operates via the protein kinase C dependent transduction pathway in the induction of class II MHC antigens on rat microvascular endothelial cells. Stimulators of protein kinase C, like PMA, replaced IFN-gamma in the induction of MHC class II on endothelial cells in a dose-dependent manner. Selective enzyme inhibitors of protein kinase C, H-7 as well as sphingosine down-regulated the IFN gamma induced class II expression in a dose-dependent manner. Addition of cAMP or cGMP in the culture, had no effect on the class II expression on the endothelial cells. Transient rise of cytosolic Ca2+ by calcium ionophore A23187, or a calmodulin antagonist W-7, had no effect on the IFN-gamma induced class II expression. PMID- 2546805 TI - Characterization of cDNA clones encoding two putative isoforms of the alpha 1 subunit of the dihydropyridine-sensitive voltage-dependent calcium channel isolated from rat brain and rat aorta. AB - cDNA clones encoding rat brain and rat aorta isoforms of the alpha 1 subunit of the dihydropyridine-sensitive, voltage-dependent calcium channel were isolated and sequenced. These tissue-specific cDNA clones share significant amino acid similarity with the rabbit skeletal muscle calcium channel alpha 1 subunit (75% and 66% amino acid identity for rat brain and rat aorta isoforms, respectively). Northern analysis revealed transcript sizes of 6.5 and 8.6 kb in aorta and 8.6 kb in brain. PMID- 2546806 TI - Conformational behavior of fragments of adrenocorticotropin and their antisense peptides determined by NMR spectroscopy and CD spectropolarimetry. AB - An 'antisense' peptide ('HTCA'), whose sequence was generated by reading the antisense RNA sequence corresponding to ACTH (1-24) was shown to bind ACTH (1-24) with a Kd of 0.3 nM in a solid-matrix binding assay [( 1986) Biochem. J. 234, 679 683]. Two-dimensional NMR spectra were used to examine the conformational behavior in methanol and in water solution of two fragments of adrenocorticotropin, ACTH(1-24) and ACTH (1-13), as well as their antisense peptides, HTCA and HTCA(12-24). The conformations are extended chains in these solutions, both as isolated molecules and when mixed with their antisense complements. The Kd values are greater than 1 mM. PMID- 2546807 TI - Regulation of colominic acid biosynthesis by temperature: role of cytidine 5' monophosphate N-acetylneuraminic acid synthetase. AB - Synthesis of colominic acid in Escherichia coli K-235 is strictly regulated by temperature. Evidence for the role of cytidine 5'-monophospho-N-acetylneuraminic acid (CMP-Neu5Ac) synthetase in this regulation was obtained by measuring its level in E. coli grown at 20 and 37 degrees C. No activity was found in E. coli grown at 20 degrees C. CMP-Neu5Ac started to be quickly synthesized when bacteria grown at 20 degrees C were transferred to 37 degrees C and was halted when cells grown at 37 degrees C were transferred to 20 degrees C. These findings suggest that temperature regulates the synthesis of this enzyme and therefore the concentration of CMP-Neu5Ac necessary for the biosynthesis of colominic acid. PMID- 2546808 TI - Calpain proteolysis of free and bound forms of calponin, a troponin T-like protein in smooth muscle. AB - Calponin, a novel homologue of troponin T, purified from chicken gizzard was found to be one of the most susceptible proteins among smooth muscle contraction associated proteins to hydrolysis by calpain I purified from human red blood cells. The high susceptibility of calponin was comparable to that reported for troponin T. The rate of degradation of calponin, unlike caldesmon and myosin light chain kinase, was accelerated when bound to calmodulin. When calponin existed as a bound form in both reconstituted actin filament and native thin filament, the rate of proteolysis was markedly retarded, indicating close association of calponin with actin filament. These observations are compatible with the view that calponin is an integral part of the actin-linked contractile machinery in smooth muscle. PMID- 2546810 TI - Developmental regulation of expression of the alpha 1 and alpha 2 subunits mRNAs of the voltage-dependent calcium channel in a differentiating myogenic cell line. AB - The voltage-dependent calcium channel (VDCC) in skeletal muscle probably plays a key role in transducing membrane charge movement to the calcium release channel. We report here that the expression of VDCC alpha 1 and alpha 2 mRNAs is developmentally regulated in differentiating C2C12 myogenic cells. The alpha 1 mRNA is not detectable in the myoblast form of C2C12 cells while its expression is induced 20-fold in differentiated myotubes. In contrast, the alpha 2 mRNA is weakly expressed in myoblasts but is also induced upon myogenic differentiation. PMID- 2546809 TI - Evidence for the existence of a cardiac specific isoform of the alpha 1 subunit of the voltage dependent calcium channel. AB - Biochemical, pharmacological and electrophysiological evidence implies the existence of tissue specific isoforms of the L-type VDCC. The alpha 1 and alpha 2 subunits of the skeletal muscle calcium channel have been previously cloned and their amino acid sequence deduced. Here we report the isolation and sequencing of a partial cDNA that encodes a heart specific isoform of the alpha 1 subunit. The amino acid sequence deduced from this part cDNA clone shows 64.7% similarity with the skeletal muscle alpha 1 subunit. Northern analysis reveals 2 hybridizing bands, 8.5 and 13 kb, in contrast to one 6.5 kb band in the skeletal muscle. Selective inhibition of mRNA expression in Xenopus oocytes by complementary oligodeoxy-nucleotides derived from the heart clone provides further evidence that the cDNA corresponds to an essential component of the VDCC. These data further support the existence of tissue-specific isoforms of the L-type VDCC. PMID- 2546811 TI - Immunodetection of Na,K-ATPase alpha 3-isoform in renal and nerve tissues. AB - At least three types of mRNA of the catalytic subunit of Na,K-ATPase namely alpha ,alpha+- and alpha 3-isoforms are identified in different tissues. Only two of them alpha and alpha+ have well known structural and catalytic properties. Here we present immunochemical data indicating that the alpha 3 protein really exists in pig and human kidney, and human brain. Crude membrane fractions and purified membrane-bound Na,K-ATPases were immunoblotted with alpha 3-specific antibodies raised against the synthetic peptide corresponding to the unique sequence of this isoform. The mature alpha 3-subunit is shown to include the sequence GDKKDDKSSPK followed by the initiating methionine residue. Nephron collecting tubules are proposed to specifically contain Na,K-ATPase alpha 3-isoform. PMID- 2546812 TI - An improved procedure for identifying and quantitating protein phosphatases in mammalian tissues. AB - The type 2A protein phosphatases in mammalian tissue extracts are inhibited completely and specifically by 1-2 nM okadaic acid. In contrast, type 1 protein phosphatases are hardly affected at these concentrations, complete inhibition requiring 1 microM okadaic acid. These observations have been exploited to develop an improved procedure for the identification and quantitation of type 1, type 2A and type 2C protein phosphatases in tissue extracts. PMID- 2546813 TI - Remarkable similarities between yeast and mammalian protein phosphatases. AB - Protein phosphatase activities in extracts of the yeast Saccharomyces cerevisiae showed remarkable similarities to the mammalian type 1, type 2A and type 2C enzymes. Similarities included their substrate specificities, including selectivity for the alpha-and beta-subunits of muscle phosphorylase kinase, sensitivity to okadaic acid and to mammalian inhibitor 1 and inhibitor 2, and requirement for divalent cations. The results suggest that the function and regulation of these enzymes has been highly conserved during evolution and indicate that the improved procedure for identifying and quantitating protein phosphatases [(1989) FEBS Lett. 250,000,000] may be applicable to all eukaryotic cells. PMID- 2546814 TI - Retinoic acid enhances VIP receptor expression and responsiveness in human neuroblastoma cell, SH-SY5Y. AB - Retinoic acid (RA) induces partial differentiation of neuroblastoma (NB) cells in vitro. In the human NB line, SH-SY5Y (a neuroblastic subclone of SK-N-SH), RA was previously shown to enhance the stimulatory (PGE1) and inhibitory (opioid) regulation of adenylyl cyclase. Since these cells are also sensitive to cAMP stimulation by vasoactive intestinal peptide (VIP), we have tested the effects of RA on VIP receptor expression and function. Pretreatment of SH-SY5Y cells with 10 microM RA over 6 days dramatically increased VIP receptor number from approximately 3,000 to approximately 70,000 sites per cell and enhanced threefold the cAMP accumulation after external VIP addition, while VIP immunoreactive content in the cells increased 2-3-fold. In the light of the recently proposed autocrine function of VIP in this cell lineage, the strong enhancement of the VIP system may contribute to the differentiation effects of RA. PMID- 2546815 TI - Specific binding of okadaic acid, a new tumor promoter in mouse skin. AB - The tumor promoter okadaic acid binds specifically to a particulate as well as a cytosolic fraction of various mouse tissues, e.g., skin, brain, lung and colon. The KD value was 21.7 nM for receptors in the particulate fraction and 1.0 nM for those in the cytosolic fraction of mouse skin. The specific binding of [3H]okadaic acid to the particulate fraction of mouse skin was inhibited dose dependently by okadaic acid, but not okadaic acid tetramethyl ether, an inactive compound, or by other tumor promoters, such as 12-O-tetradecanoylphorbol-13 acetate and teleocidin. The results suggest a new pathway of tumor promotion mediated through the okadaic acid receptor(s). PMID- 2546816 TI - Nucleoside diphosphate kinase from human erythrocytes: purification, molecular mass and subunit structure. AB - A new procedure for the purification of nucleoside diphosphate kinase from human erythrocytes is described. The enzyme (105 kDa by gel filtration) is made-up of two different kinds of subunits (19.0 and 20.5 kDa), both displaying enzymatic activity. The probable subunit structure of the enzyme is hexameric. The discrepancies related to earlier work are discussed. PMID- 2546817 TI - An analysis of the periodicity of conserved residues in sequence alignments of G protein coupled receptors. Implications for the three-dimensional structure. AB - Twenty-three sequences from the family of G-protein coupled receptors have been aligned according to the 'historical alignment' procedure of Feng and Doolittle. Fourier transform analysis of this reveals that parts of five of the seven putative membrane-spanning regions exhibit a periodicity of conserved/nonconserved residues which is compatible with the periodicity of the alpha-helix. This would place the conserved residues on one side of the helix, which may face the inside of the proposed seven membered helical bundle. PMID- 2546818 TI - Control of fructose 2,6-bisphosphate levels in rat macrophages by glucose and phorbol ester. AB - The presence of fructose 2,6-bisphosphate (Fru 2,6-P2) in elicited peritoneal macrophages of rat was examined. These cells possess an active phosphofructokinase-2 which is diminished by citrate and only slightly inhibited by glycerol 3-phosphate. Phosphofructokinase-1 submaximal activity was increased 26-fold by the addition of 1 microM Fru 2,6-P2. Incubation of cells without glucose decreased the amount of Fru 2,6-P2 to zero, but further addition of 5 mM glucose increased the levels of the sugar ester 20-fold. In addition, the presence of phorbol ester potentiated the synthesis of Fru 2,6-P2. By contrast phenylisopropyladenosine or prostaglandin F2 alpha inhibited the production of Fru 2,6-P2. PMID- 2546819 TI - In vitro transcription of the mouse whey acidic protein promoter is affected by upstream sequences. AB - The promoter sequences from -175 to +10 of the mouse whey acidic protein (WAP) gene are a target for sequence-specific binding of nuclear proteins [(1987) Nucleic Acids Res. 15, 2103-2121]. Using in vitro transcription assays based on nuclear extracts, transcription factors were shown to bind to these sequences and promoter upstream sequences were found to stimulate transcription. PMID- 2546820 TI - The cDNA of the two isoforms of bovine cGMP-dependent protein kinase. AB - cDNAs encoding the isoform I alpha of the cGMP-dependent protein kinase were isolated from a bovine trachea smooth muscle cDNA library constructed in lambda gt10. The deduced protein sequence is identical with the protein sequence obtained by Edman degradation of the bovine lung enzyme [(1984) Biochemistry 23, 4207-4218]. Alternate cDNA clones were isolated which code for a protein slightly different within the aminoterminal part from the known amino acid sequence. These alternate cDNAs contain the sequence of a peptide identified in the isoform I beta of cGMP-dependent protein kinase. Northern blot analysis of poly(A)+ RNA from bovine trachea smooth muscle indicated the presence of two different mRNA species of about 6.2 kb. PMID- 2546821 TI - Lipocortin I and lipocortin II inhibit phosphoinositide- and polyphosphoinositide specific phospholipase C. The effect results from interaction with the substrates. AB - Lipocortins I and II, known to inhibit phospholipase A2, have been purified from bovine lung and tested with respect to their ability to affect the enzymatic activities of phosphoinositide- and polyphosphoinositide-specific phospholipase C from human platelets, rat liver cytosol or rat brain membranes. At 0.67 microM, both lipocortins led to complete inhibition of phospholipase C activity with either phosphatidylinositol or phosphatidylinositol 4,5-bisphosphate as substrate. The inhibition could be overcome by increasing the substrate concentration. Ultracentrifugation studies with lipocortin II showed a direct interaction between phosphatidylinositol and the lipocortin, indicating that the lipocortins inhibit phospholipase C not directly but by interacting with the substrate. In experiments with plasma membranes from [3H]inositol-labeled HL-60 cells, lipocortin II did not affect PI-specific phospholipase C activity in the absence or presence of calcium plus or minus GTP-gamma-S. PMID- 2546822 TI - Purification of a p47 phosphoprotein from Xenopus laevis oocytes and identification as an in vivo and in vitro p34cdc2 substrate. AB - This paper describes the purification of a 47 kDa protein from Xenopus laevis oocytes that becomes phosphorylated when the oocytes undergo meiotic maturation. This protein (p47) is part of a high molecular mass complex containing at least two other proteins of molecular mass 30 and 36 kDa. This complex can be isolated from stage VI oocytes before maturation. We obtained a pattern for phosphopeptides in p47 phosphorylated in vivo very similar to that of the purified protein phosphorylated in vitro by p34cdc2 (a H1 kinase which is a component of the M-phase promoting factor) and [gamma-32P]ATP. Therefore, the purified p47, already described as a marker of MPF activity, is the first reported in vivo substrate for the cell division control kinase. PMID- 2546823 TI - Inhibition of lipoprotein lipase by the receptor-binding domain of apolipoprotein E. AB - A synthetic peptide (residues 139-153) corresponding to the receptor-binding domain of apolipoprotein E (ApoE) was tested for lipoprotein lipase (LPL) inhibitory properties. In systems using both natural and synthetic substrates, inhibition of LPL was observed. Using the synthetic substrate, 50% inhibition was observed at 50 microM while high concentrations completely inhibited LPL activity. These studies suggest an additional functional role for the receptor binding domain of ApoE-modulation of LPL activity. PMID- 2546824 TI - Regulation of hepatic expression of IGF I and fetal IGF binding protein mRNA in streptozotocin-diabetic rats. AB - Hepatic mRNA levels of insulin-like growth factor I (IGF I) and of the fetal, nonglycosylated 32 kDa IGF-binding protein (BP) were analysed in diabetic, diabetic insulin- and IGF I-treated rats as well as in age-matched, healthy control animals. IGF ImRNA levels are reduced in diabetic rats and increased by insulin treatment. In contrast, the infusion of IGF I does not significantly upregulate IGF I mRNA levels. Fetal IGF BP mRNA expression is very low in healthy control animals, but high levels are found in diabetic rats. Insulin therapy lowers fetal IGF BP mRNA levels, whereas IGF I has no effect. We propose that insulin is a major regulator of the 32 kDa IGF BP levels in adult rats. PMID- 2546825 TI - Appearance of a late stage during mammalian DNA replication when cells resume formation of 10 kb DNA replication intermediates. AB - After the joining of human large DNA replication intermediates and before the appearance of mature chromatin DNA, there exists a distinct stage--'the post elongation stage'. This stage reappears during recovery of DNA synthesis simultaneously with the reappearance of a large DNA replication intermediate, 10 kb DNA. PMID- 2546826 TI - Influence of non-esterified fatty acids on respiratory control of reconstituted cytochrome-c oxidase. AB - Bovine heart cytochrome-c oxidase was reconstituted in liposomes (asolectin) and the activity measured in the presence and absence of uncoupler at increasing concentrations of non-esterified fatty acids. Palmitic and stearic acids resulted in a decrease of about 40% in the respiratory control ratio at a concentration of 1 microM, when measured using a spectrophotometric procedure but not with a polarographic assay method. At higher fatty acid concentrations no further change was found. A 50% decrease in respiratory control was determined when the enzyme was reconstituted in pure phosphatidylcholine containing 2% cardiolipin. The respiratory control of reconstituted cytochrome-c oxidase from bovine liver was not influenced by fatty acids. PMID- 2546827 TI - Na+/H+ exchange in mitochondria as monitored by BCECF fluorescence. AB - The recently developed method of loading isolated heart mitochondria with the fluorescent pH indicator, BCECF, was applied to monitor the Na+o/H+i exchange process from the matrix side of the membrane. The Na+-induced changes in the pH of the matrix (pHm) showed that: (i) the Na+o/H+i exchange followed Michaelis Menten kinetics with respect to external Na+ with a Km of approx. 20 mM; (ii) in contrast to this, the dependence of the exchange rate on the matrix [H+] did not obey the Michaelian model. No Na+-induced alkalinization occurred above a pHm of 7.45 +/- 0.09 (n = 4). Below this value the reciprocal of the transport rate and that of the matrix [H+] deviated upwardly from the straight line. The results suggest that internal H+ might exert allosteric control on the mitochondrial Na+/H+ exchange process. PMID- 2546828 TI - Human nuclear protein interacting with a conservative sequence motif of Alu family DNA repeats. AB - Human retrotransposons, Alu-family DNA repeats (AFRs), have variable nucleotide sequence but conservative short elements, which may have important functions, are also present. In our previous reports we have described human nuclear DNA-binding protein interacting with AFRs and evidence was presented that the protein recognizes sequence motif 5'-GGAGGC-3' which is conserved in the spacer of RNA polymerase III promoter of AFRs and in the SV40 T-antigen-dependent replication origin of AFRs. In this study it was found that double-stranded synthetic oligonucleotides containing indicated conservative sequences of AFRs actually have high-affinity binding site for HeLa nuclear protein. The data suggest that non-infected human cells contain nuclear DNA-binding protein which recognizes the conservative sequence motif of AFRs - GGAGGC. PMID- 2546829 TI - A model for the ability of drugs to induce enhanced DNase I cleavage. AB - A common property of sequence-selective DNA-binding drugs lies in their ability to induce an enhanced DNase I cleavage in regions surrounding their binding sites. A hypothetical model to explain the enhancements induced by drug binding to the minor-groove of DNA is presented. It involves the participation of three different single models: a mass action effect produced by the enzyme redistribution after drug binding; changes in the minor groove width size; and interactions between the enzyme and the drug, so increasing the cleavage in places located close to the binding site. The model is tested by using statistical data analysis. The hypothetical model might explain the experimental results better than any of the single models alone, but these models also appear to render significant results. PMID- 2546830 TI - The primary structure of rat ribosomal protein L26. AB - The amino acid sequence of rat ribosomal protein L26 was deduced from the sequence of nucleotides in a recombinant cDNA and confirmed from the NH2-terminal amino acid sequence of the protein. Rat L26 contains 145 amino acids and has a molecular mass of 17,266 Da. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 8-16 copies of the L26 gene. The mRNA for the protein is about 650 nucleotides in length. Protein L26 has a sequence of 9 residues that may be repeated in three places. PMID- 2546832 TI - Fine needle aspiration of the lung at the Medical Center of Delaware: a retrospective review of 243 patients. PMID- 2546831 TI - Effects of pretreatment with tetradecanoyl phorbol acetate on regulation of growth hormone and prolactin secretion from ovine anterior pituitary cells. AB - Tetradecanoyl phorbol acetate (TPA) stimulates growth hormone (GH) and prolactin secretion from ovine anterior pituitary cells. Pretreatment of the cells with TPA abolishes this effect, presumably due to down-regulation of protein kinase C. Such pretreatment did not alter effects of thyrotropin-releasing hormone or dopamine on prolactin secretion, suggesting no involvement of protein kinase C. Pretreatment with TPA attenuated actions of GH-releasing hormone on GH release (but not actions on cyclic AMP levels), possibly due to depletion of cellular stores of GH. Such pretreatment also attenuated inhibition of GH release by somatostatin, possibly due to phosphorylation of receptors or associated proteins by protein kinase C. PMID- 2546834 TI - Sodium-n-butyrate induces secretion and substrate accumulation of p52 in Kirsten sarcoma virus-transformed rat kidney fibroblasts. AB - 1. A group of five transformation-responsive secreted proteins, ranging in molecular mass from 31 to 70 kDa, were identified in cultured normal rat kidney (NRK) fibroblasts. 2. One such protein (p52) was found to be a major secreted and substrate-attached component of NRK cells. 3. Kirsten sarcoma virus-transformed NRK cells failed to accumulate p52 in either the secreted or substrate-associated protein compartments; this protein was inducible, however, in transformed cells by culture in 2 mM sodium-n-butyrate. 4. Kinetics of p52 induction in transformed NRK cells, relative to the time course of increased cell spreading, and its enrichment in the substrate-associated protein fraction suggest that p52 might function in cell-substrate attachment. PMID- 2546833 TI - Characterization of hormone-sensitive adenylate cyclase in rabbit gallbladder mucosa. AB - 1. We have developed a plasma membrane preparation from the mucosal epithelium of rabbit gallbladder and have characterized the hormonal sensitivity of adenylate cyclase in this preparation. 2. Basal activity is low and is stimulated by GTP and GppNHp. Hormonal stimulation is largely dependent on exogenous guanine nucleotide. 3. Several prostaglandins (E1 approximately E2 greater than A1 greater than B1), vasoactive intestinal peptide and the beta-adrenergic agonist, isoproterenol, stimulate mucosal adenylate cyclase activity; a variety of peptides and neurotransmitters (secretin, cholecystokinin, arg-vasopressin, oxytocin, histamine, dopamine and serotonin) are without effect. 4. The data support the hypothesis that the inhibitory effect of prostaglandins, vasoactive intestinal peptide, and isoproterenol on gallbladder fluid absorption in certain species may be mediated by cyclic AMP. 5. The membrane preparation should be useful in further characterizing hormone receptor-transducer interactions of the gallbladder mucosal epithelium. PMID- 2546835 TI - Studies of blocking mechanisms of carbachol-induced polyphosphoinositide turnover in rat cortical synaptosomes by neuroactive drugs. AB - 1. The incorporation of 32Pi into 4 phospholipids of rat cortical synaptosomes was altered in the presence of carbachol (1 mM), viz. a decrease of phosphatidylinositol-4,5-bisphosphate and phosphatidyl-4-phosphate by 34 and 21%, and an increase of phosphatidylinositol and phosphatidic acid by 52 and 96% of basal controls respectively. 2. The IC30 values calculated from the dose-response curves for drugs affecting carbachol-induced 32Pi incorporation into these phospholipids, and [3H]QNB binding to the cortical synaptosomes were similar for the typical antimuscarinic agents (i.e. atropine, pirenzepine and trihexyphenidyl), and tricyclic antidepressants (i.e. amitriptyline, doxepin and imipramine) studied. 3. The IC30 values obtained for drugs affecting carbachol induced 32Pi incorporation into these phospholipids, and high potassium-induced 45Ca2+-uptake by this preparation were similar for neuroselective calcium channel blockers (i.e. cinnarizine and flunarizine) studied. 4. Our results suggest that the neuroactive drugs studied can either act at, or beyond the receptor level, perhaps on the availability of calcium ion, to block carbachol-induced polyphosphoinositide turnover in rat cortical synaptosomes. PMID- 2546836 TI - Twenty-fifth Colworth medal lecture. The cellular functions of myo-inositol. PMID- 2546837 TI - Lipids and multiple sclerosis. PMID- 2546838 TI - Detection of proton-linked sugar transport proteins in Enterobacteriaceae. PMID- 2546839 TI - Effect of lipoxygenase products on choriogonadotropin secretion by cultured human choriocarcinoma JEG-3 cells pre- and post-stimulation with epidermal growth factor and a phorbol ester. AB - Previous studies using arachidonic acid and preferential inhibitors of the arachidonic acid pathway have implicated the lipoxygenase system in choriogonadotropin (hCG) secretion by JEG-3 cells. Presently, JEG-3 cells are used in order to examine the effect of lipoxygenase products on hCG secretion. Results show that 30 microM 15-hydroxyeicosatetraenoic acid (15-HETE) induces an approximately 3-fold increase in basal hCG secretion, while 5-HETE, 12-HETE, and leukotriene LTA4 have no significant effect. In addition, 15-HETE potentiates the stimulation of hCG secretion induced by 3 nM epidermal growth factor (EGF), but has no significant effect on the stimulation of hCG induced by 22 nM tetradecanoylphorbol acetate (TPA). The present study further implicates the arachidonic acid pathway in the control of hCG secretion and documents that the effect of EGF can be rate-limited by a product of the lipoxygenase system. PMID- 2546840 TI - Isolation and characterisation of calcineurin from adrenal cell cytoskeleton: identification of substrates for Ca2+-calmodulin-dependent phosphatase activity. AB - Ca2+-calmodulin-dependent protein phosphatase activity is found in cytoskeletons of Y-1 mouse adrenal and bovine fasciculata cells. The activity is inhibited by three inhibitors of calmodulin (trifluoperazine, W-7 and pimozide) with EC50 in the low micromolar range. Protein phosphatase activity is inhibited by vanadate, fluoride, Zn2+ and pyrophosphate, stimulated by Mn2+ and found to be tightly bound to the cytoskeleton. Substrates for endogenous phosphatase activity were defined by one- and two-dimensional polyacrylamide gels. Phosphatase activity was seen with proteins that are substrates for both cyclic AMP-dependent and cyclic AMP-independent kinase enzymes. One specific Ca2+-calmodulin-dependent phosphatase, namely calcineurin, was purified to near homogeneity from cytoskeletons of Y-1 cells. The enzyme was found to be a heterodimer (MW 61,000 and 16,000) and the smaller subunit was shown to cross-react with antibodies raised against calcineurin from bovine brain. The purified enzyme catalyzes dephosphorylation of proteins (phosphorylase kinase and casein), phosphoamino acids (tyr greater than thre greater than ser) and a synthetic substrate (p nitrophenyl phosphate). In addition, a new application of membrane transfer was devised by which the purified enzyme was incubated with a Western blot of cytoskeleton following incubation with [32P]ATP. This method defined four specific substrates of the enzyme (MW 150,000, 55,000, 35,000 and 30,000). Anti calcineurin revealed that only a single Ca2+-calmodulin-dependent phosphatase is found in adrenal cell cytoskeleton.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546841 TI - Developmental changes in phosphodiesterase activity and hormonal response in the prothoracic glands of Manduca sexta. AB - Prothoracicotropic hormone (PTTH) stimulates ecdysteroid secretion by the prothoracic glands of Manduca sexta in a cAMP-dependent manner. However, larval and pupal glands differ markedly in the degree to which PTTH stimulates cAMP accumulation, suggesting a stage-specific difference in phosphodiesterase activity. The present study was designed to determine if and when such a difference arose during development, and its effect on PTTH-stimulated ecdysteroid secretion. The results reveal that soluble phosphodiesterase activity in the prothoracic glands changes significantly during the course of the fifth (last) larval instar, with a marked increase in activity occurring at the onset of prepupal development. Phosphodiesterase activity, particularly in the soluble cell fraction, is inversely correlated with PTTH-stimulated cAMP accumulation. Hormone-stimulated ecdysteroid secretion does not require cAMP accumulation, but does appear to require detectable cAMP synthesis as measured in the presence of phosphodiesterase inhibitors. The amount of ecdysteroid secreted, however, is not proportional to the amount of cAMP synthesized but rather is more closely correlated with developmental changes in glandular protein content. PMID- 2546842 TI - Adrenocorticotropic hormone inhibits angiotensin II-stimulated inositol phosphate accumulation in rat adrenal glomerulosa cells. AB - Rat adrenal glomerulosa cells labelled for 18 h with [3H]inositol responded to angiotensin II with a dose-dependent stimulation of the accumulation of inositol monophosphate, inositol bisphosphate and inositol trisphosphate. Addition of adrenocorticotropic hormone (ACTH) (10(-7)M) reduced the maximum responses without altering the EC50 values for angiotensin II. Thus, ACTH acted as a non competitive inhibitor with respect to angiotensin II. No inhibition was observed in cells labelled for 2 h with [3H]inositol. Detailed examination of the inhibition showed that ACTH(1-24) was the most potent inhibitor, with ACTH(1-39) being 10-fold less potent. A mixture of alpha-melanocyte-stimulating hormone (alpha-MSH) (ACTH(1-13] and corticotropin-like intermediate lobe peptide (ACTH(18 39] was similarly inactive. ACTH(5-24) did not produce detectable inhibition. In terms of specificity, the receptor mediating ACTH inhibition of phosphatidylinositol turnover was similar to the receptor which mediated stimulation of aldosterone synthesis. Inhibition by ACTH was additive with inhibition produced by dibutyryl cAMP demonstrating that it was not mediated by rises in intracellular cAMP. ACTH inhibition also was additive with inhibition by the calcium channel blocker, nifedipine. These results demonstrate an interaction between ACTH receptors and angiotensin II receptors in adrenal glomerulosa cells at the level of their receptor-second messenger pathways. PMID- 2546843 TI - Influence of insulin on phosphate uptake by brush border membranes from human placenta. AB - Regulation of phosphate transport by insulin was investigated in brush border membranes from human placenta at term. At 22 degrees C, a 45 min incubation of the total tissue with 10(-6) M insulin significantly decreased both the initial rate and the peak of sodium-dependent phosphate uptake by the corresponding brush border membranes. In contrast, Na+ transport was not influenced by the hormone. Increasing the insulin concentration from 0 to 10(-5) M resulted in a dose dependent inhibition of phosphate uptake with half-maximal effect at 1.1 x 10(-9) M. The hormone decreased PO4 transport by decreasing the affinity of the carrier for the substrate (Km = 0.180 +/- 0.010 mM and 0.215 +/- 0.015 mM in absence and presence of 10(-6) M insulin respectively, P less than 0.05). The inhibitory effect of insulin required the presence of Mn2+ whereas neither Mn2+ nor insulin alone had any influence on PO4 uptake. It is therefore assumed that receptor phosphorylation, which needs the presence of Mn2+, is an intermediate step of insulin action on PO4 uptake by the subsequently isolated brush border membranes. In contrast, insulin had no effect on PO4 uptake when the membranes were directly incubated with the hormone prior to the transport measurement, suggesting that an intracellular messenger is needed for the inhibitory effect. This messenger is not cAMP since insulin at 10(-6) M concentration has no effect on cAMP content of the total placental tissue.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546844 TI - Effect of galactose and glucose levels and sorbinil treatment on myo-inositol metabolism and Na+-K+ pump activity in cultured neuroblastoma cells. AB - Neuroblastoma cells were used to analyze the effect of galactose supplementation on myo-inositol metabolism, polyol accumulation, and Na+-K+ pump activity. Culturing cells in 30 mM galactose for a minimum of 1 wk led to a large accumulation of intracellular galactitol and a greater than 50% decrease in myo inositol content. The effect of galactose on the intracellular content of galactitol and myo-inositol was concentration dependent. Extracellular myo inositol accumulation and incorporation into phospholipid decreased by 20-30% in cells grown in 30 mM galactose. The decrease in myo-inositol accumulation is apparently due to a noncompetitive inhibition of high-affinity myo-inositol uptake. Treatment of the galactose-containing media with 0.4 mM sorbinil partially prevented the galactose-mediated decreases in myo-inositol metabolism and content. The galactitol content of the sorbinil-treated cells was significantly reduced compared with the galactitol levels in cells cultured in 30 mM galactose; however, galactitol levels remained significantly elevated over control cells. Exposing neuroblastoma cells to 30 mM galactose causes a decrease in the levels of phosphatidylinositol that is partially restored by the addition of sorbinil. The activity of the Na+-K+ pump was decreased by 20% in cells cultured in 30 mM galactose and was partially protected by sorbinil treatment. The effects of long-term galactose supplementation on myo-inositol metabolism, polyol accumulation, and Na+-K+-ATPase transport activity in cultured neuroblastoma cells are similar to the effects of high concentrations of glucose. These results provide additional evidence that the accumulation of polyol by neuroblastoma cells is partially responsible for alterations in myo-inositol metabolism and decreases in Na+-K+-ATPase transport activity. PMID- 2546845 TI - Hypoglycaemia induced by antibodies to insulin receptor following a bone marrow transplantation in an immunodeficient child. AB - Severe hypoglycaemia developed seven months after a bone marrow transplantation in a child with severe combined immunodeficiency. His serum exerted potent insulin-like activity: (a) it stimulated insulin receptor autophosphorylation and kinase activity in cell-free systems, this effect being additive to insulin; (b) it increased glucose transport in isolated soleus muscle. These insulin-like effects were due to immunoglobulins against the insulin receptor. Indeed, the patient serum immunoprecipitated human or murine insulin receptors from different tissues and inhibited insulin binding to receptor on human IM-9 lymphocytes. After corticoids and immunosuppressive therapy by azathioprine, the patient hypoglycaemic episodes disappeared, and concomitantly, the antibodies to insulin receptor were no longer detected, as judged by both immunoprecipitation of insulin receptor and stimulation of glucose transport. PMID- 2546846 TI - Enalapril retards glomerular basement membrane thickening and albuminuria in the diabetic rat. AB - This study has evaluated the effects of the angiotensin converting enzyme inhibitor Enalapril on glomerular ultrastructure and albuminuria in normotensive and hypertensive diabetic rats. Streptozotocin-diabetes was induced in Wistar Kyoto and spontaneously hypertensive rats. Enalapril was administered in drinking water in diabetic normotensive, control hypertensive and diabetic hypertensive rats. Enalapril therapy prevented an increase in glomerular basement membrane thickness in diabetic normotensive, control hypertensive and diabetic hypertensive rats without any significant effect on fractional mesangial volume. Enalapril decreased albuminuria in diabetic normotensive, control hypertensive and diabetic hypertensive rats. Thus, enalapril retards the development of glomerular basement membrane thickening and albuminuria in the rat, in the presence or absence of hypertension. PMID- 2546847 TI - G proteins control diverse pathways of transmembrane signaling. AB - Hormones, neurotransmitters, and autacoids interact with specific receptors and thereby trigger a series of molecular events that ultimately produce their biological effects. These receptors, localized in the plasma membrane, carry binding sites for ligands as diverse as peptides (e.g., glucagon, neuropeptides), lipids (e.g., prostaglandins), nucleosides and nucleotides (e.g., adenosine), and amines (e.g., catecholamines, serotonin). These receptors do not interest directly with their respective downstream effector (i.e., an ion channel and/or an enzyme that synthesizes a second messenger); rather, they control one or several target systems via the activation of an intermediary guanine nucleotide binding regulatory protein or G protein. G proteins serve as signal transducers, linking extracellularly oriented receptors to membrane-bound effectors. Traffic in these pathways is regulated by a GTP (on)-GDP (off) switch, which is regulated by the receptor. The combination of classical biochemistry and recombinant DNA technology has resulted in the discovery of many members of the G protein family. These approaches, complemented in particular by electrophysiological experiments, have also identified several effectors that are regulated by G proteins. We can safely assume that current lists of G proteins and the functions that they control are incomplete. PMID- 2546848 TI - Plasticity of neuronal receptors. AB - This article describes ways in which receptors, key components of signal propagation through a synapse, can mediate changes in that propagation. Changes occur at four levels: in the signal-transducing capability of a single receptor molecule, in the number of receptors per cell, in the subcellular placement of receptor molecules, and in the cytoarchitecture of receptor-rich regions. The ability of receptors to shift between different desired states is called plasticity, and such shifts can be long-lived as well as transient. In this article we focus on neuronal receptors, although key findings from a variety of cell systems are reported. Neuronal receptor plasticity may have a special role in the assembly as well as the adaptability of the nervous system. PMID- 2546849 TI - [Myocardial scintigraphy using 99m-technetium pyrophosphate in the diagnosis of acute myocardial infarct. Comparison of planar and tomographic imaging]. AB - The aim of the study was to compare the effectiveness of planar and single photon emission computed tomography (SPECT) imaging with 99mTc-pyrophosphate in the verification of acute myocardial infarction. The study was performed on 39 patients (26 males, 13 females) aged between 41-76 years (mean 61 +/- 9) admitted to CCU for acute myocardial infarction. Patients underwent planar and SPECT imaging using a double-head-camera with a 360 degrees rotation arc within 6 days of admission. Planar images were obtained in three standard projections (anterior, LAO 45 degrees, LAO 75 degrees) with acquisitions of 500000 counts each. To obtain SPECT images ninety projections, each lasting 20 seconds, were taken; subsequently images reconstruction was performed using an iterative algorithm. Positive planar images were found in 21 out of 39 patients; SPECT images were positive in 33 patients. In "Q wave" myocardial infarctions planar images were positive in 13 out of 15 patients, whereas SPECT images were positive in all subjects; in "non-Q wave" myocardial infarctions planar images were positive in 6 out of 22 patients whereas SPECT images were positive in 16 patients (p less than 0.005); in 2 patients with left bundle branch block both planar and SPECT images were positive. A bidimensional echocardiogram was carried out on 38 patients: an alteration of left ventricular regional wall motion was present in 30 subjects; in one patient diffuse hypokinesia was present.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2546850 TI - Effects of the benzodiazepine receptor antagonist flumazenil in hepatic encephalopathy in humans. AB - If increased gamma-aminobutyric acid (GABA)-mediated neurotransmission contributes to the mediation of hepatic encephalopathy, it may be possible to induce ameliorations of the syndrome by pharmacologically antagonizing a component of the GABA/benzodiazepine receptor complex. To test this possibility we administered the benzodiazepine receptor antagonist flumazenil by intravenous injection to 14 patients with hepatic encephalopathy complicating cirrhosis. Flumazenil administration induced variable and transient, but distinct, improvements of the mental status in 71% of the patients. The degree of encephalopathy improved from stage IV to stage II in 4 patients and from stage IV to stage III in 2 patients. The mental status of all patients with less advanced encephalopathy (3 with stage III, 1 with stage II) also improved, but these responses were clinically less impressive. The arousal effect occurred within minutes after the injection and lasted for 1 to 2 h. Furthermore, it was associated with a significant increase of the mean electroencephalographic frequency from 4.2 to 5.2 cycle/s. Of the 8 patients who were ultimately discharged from the hospital, 7 had responded to flumazenil. No patient who died within 48 h of receiving flumazenil had shown any arousal effect. These findings strongly favor a prominent pathogenetic role of increased GABAergic tone in hepatic encephalopathy in humans and suggest that a positive response to flumazenil might be of prognostic value in predicting short-term survival in encephalopathic patients with liver disease. PMID- 2546851 TI - Hepatic injury related to enalapril. PMID- 2546852 TI - Beta-endorphin and ACTH levels in the perioperative period. AB - 1. The purpose of this study was to determine the plasma levels of beta-endorphin (beta-END) and ACTH in the perioperative period, define correlations of hormonal plasma levels with clinical parameters and establish the effect of droperidol premedication on hormonal levels and clinical parameters. 2. Twenty two were assigned to one of two groups: (1) Control (no premedication) and (2) droperidol (7.5 mg im) premedication. 3. Venous blood samples and clinical evaluations were done the day prior to surgery, just prior to induction of anesthesia and 1-1.5 hr postoperatively. 4. The results indicate that (1) expectancy of surgery on arrival to the operating room increases beta-END but not ACTH plasma levels, (2) this increase in beta-END is not affected by droperidol administration and (3) postsurgical stress increases beta-END and ACTH above operating room levels. 5. These results indicate that although beta-END and ACTH are both produced by the pituitary and derived from a common precursor, the type of stimuli (pre- versus postsurgical stress) seems to differentially affect their plasma levels. PMID- 2546853 TI - The effects of some possible inhibitors of ectonucleotidases on the breakdown and pharmacological effects of ATP in the guinea-pig urinary bladder. AB - 1. The effects of some possible inhibitors of ectonucleotidases on the breakdown of extracellular ATP by strips of guinea-pig urinary bladder were investigated. 2. Suramin and ethacrynic acid (10 mM) both inhibited ATP breakdown significantly, and difluorodinitrobenzene (10 mM) inhibited it slightly whereas N ethylmaleimide, adenosine 5'-(gamma-thiotriphosphate) (ATP-gamma-S) and reactive blue-2 (10 mM) were without effect. 3. The inhibitory effects of suramin on ATP breakdown were non-competitive. 4. Ethacrynic acid (1 mM) irreversibly inhibited contractions of the guinea-pig bladder induced by ATP, substance P, histamine, non-adrenergic, non-cholinergic nerve stimulation or KCl, whereas suramin (100 microM) had no inhibitory effect. 5. The results suggest that suramin might provide a starting point for the design of selective inhibitors of ectonucleotidases. PMID- 2546854 TI - Suggestive evidence for inhibitory action of amiodarone on Na+, K+-pump activity in guinea pig heart. AB - 1. Effects of amiodarone, a powerful antiarrhythmic agent, on Na+, K+-pump activity were examined on ventricular papillary muscle of guinea pig, by means of conventional microelectrode technique. 2. The activity of Na+, K+-pump was measured by two methods. One of them was to measure the amplitudes of depolarization observed during overdrive stimulation (3.3 Hz) and of hyperpolarization observed after the overdrive stimulation (post-overdrive hyperpolarization), and the other, to measure the hyperpolarization observed following introduction of 10 mM K+, after exposure to K+ free solution for a certain duration. 3. Amiodarone significantly decreased the amplitude of depolarization during overdrive stimulation and the amplitude of post-overdrive hyperpolarization. 4. In the latter method, the deactivation process of hyperpolarization recorded by the introduction of 10 mM K+ following K+ depletion slowed down by amiodarone. 5. These findings suggest that amiodarone may inhibit, at least in part, the Na+, K+-pump activity in the ventricular muscle of guinea pig. PMID- 2546855 TI - Interaction of thymic peptide thymosin alpha 1 with VIP receptors in rat intestinal epithelial cells: comparison with PHI and secretin. AB - 1. Thymic peptide thymosin alpha 1 (10(-10)-10(-7) M) is shown to inhibit the specific binding of 125I-labelled VIP. 2. Thymosin alpha 1 was 1500 times less potent than VIP at inhibiting 125I-labelled VIP binding. 3. Thymosin alpha 1 (10( 10)-10(-7) M) was weak in stimulating cyclic AMP production. 4. Interaction of thymosin alpha 1 with VIP receptors is compared with PHI and secretin. 5. The order of affinity of different peptides is VIP greater than PHI greater than secretin greater than thymosin alpha 1. PMID- 2546856 TI - Characterization of in vitro constructed IS30-flanked transposons. AB - In order to facilitate functional studies on the mobile genetic element IS30, a resident of the Escherichia coli chromosome, transposon structures with two copies of IS30 flanking the chloramphenicol-resistance gene cat were constructed in vitro. Transposons containing IS30 as direct repeats (Tn2700 and Tn2702) transpose from multicopy plasmids into the genome of phage P1-15, thus giving rise to special transduction for cat with frequencies between 10(-5) and 10( 8)/plaque-forming unit. In contrast, transposon structures with IS30 in inverted repeat (Tn2701 and Tn2703) showed no detectable (less than 10(-9] transposition activity in vivo. By restriction analysis, two insertion sites of Tn2700 and Tn2702 on the phage P1-15 genome were indistinguishable from those observed earlier with a single copy of the IS30 element. These two insertion sites were used several times independently by Tn2700 and Tn2702. This confirms the non random target selection by the element and it indicates that transposition of Tn2700 and Tn2702 follows the same rules as that of IS30. PMID- 2546857 TI - IS257 from Staphylococcus aureus: member of an insertion sequence superfamily prevalent among gram-positive and gram-negative bacteria. AB - The nucleotide sequences for the IS257 family of insertion sequences from Staphylococcus aureus were compared with those of the ISS1 family from Streptococcus lactis and the IS15 family which is widespread amongst Gram negative bacteria. These elements have a striking degree of similarity in both their putative transposase polypeptide sequences and their nucleotide sequences (40 to 64% between pairs), including 12 out of 14 bp conservation in their terminal inverted repeats. The evolutionary distance between the IS15 family and the IS257 and ISS1 families of Gram-positive origin is approximately twice that between the IS257 and ISS1 families. Analysis of base substitutions in the three sequences has provided insights into the effect of selection for the G + C content of immigrant genes to conform to that of their hosts, and into the evolution of biases in overall amino acid composition of cellular proteins in prokaryotes and eukaryotes. The IS257, ISS1, IS15 families form a superfamily of insertion sequences that has been involved in the spread of a number of antimicrobial resistance determinants in Gram-positive and Gram-negative pathogens. PMID- 2546858 TI - Factors affecting transposition activity of IS50 and Tn5 ends. AB - The partially matched I and O ends of IS50 (the insertion sequence of the transposon Tn5) are needed for transposition, probably as the sites upon which the cis-acting transposase and host proteins act. To better understand how transposition is regulated we made a series of IS50-related elements in which the positions of the ends and of the transposase gene were varied systematically. Assays of these elements showed that the I and O ends differ inherently in transposition activity. Other workers showed that methylation, at DNA N6-adenine methyltransferase (Dam) recognition sites within the I end and the transposase tnp gene promoter, inhibits transposase synthesis and also I end activity. We show that the effect of Dammediated methylation on an I end depends on the end's orientation relative to the tnp gene. Further, in dam+ cells oriented like -tnp-- -in relation to the first and second ends) are (O, I) greater than (O, O) greater than or equal to (I, O) greater than (I, I). In dam- cells the relative activities are (O, I) = (I, O) = (I, I) greater than (O, O). Our results are consistent with a model orginally developed for IS10, in which hemi-methylation resulting from passage of a replication fork regulates transposition. PMID- 2546859 TI - Omegon-Km: a transposable element designed for in vivo insertional mutagenesis and cloning of genes in gram-negative bacteria. AB - To combine the features of the omega interposons with the advantages of in vivo transposition mutagenesis, we have constructed an artificial transposon, called Omegon-Km. The Omegon-Km transposon is carried on the plasmid pJFF350 which can be conjugally mobilized into a broad range of Gram-negative bacteria. Omegon-Km is flanked, in inverted orientation, by synthetic 28-bp repeats derived from the ends of IS1. In addition, each end of Omegon-Km has the very efficient transcription and translation terminators of the omega interposon. Internally, Omegon-Km carries the selectable kanamycin (Km)-neomycin resistance gene (alph A) which is expressed well in many Gram-negative bacteria. The IS1 transposition functions are located on the donor plasmid but external to Omegon-Km. Thus, insertions of Omegon-Km are very stable because they lack the capacity for further transposition. Omegon-Km mutagenesis is performed by conjugal transfer of pJFF350 from Escherichia coli into any Gram-negative recipient strain in which this plasmid is unable to replicate. Those cells which have had a transposition event are selected by their resistance to Km. Very high frequencies of Omegon-Km transposition were observed in Pseudomonas putida. Preliminary experiments with other Gram-negative soil and water bacteria (Rhizobium leguminosarum, Paracoccus denitrificans) yielded mutants at reasonable levels. The presence of an E. coli specific origin of replication (ori) within Omegon-Km allows the rapid and easy cloning, in E. coli, of the nucleotide sequences flanking the site of the transposition event. PMID- 2546860 TI - Characterization and nucleotide sequence determination of a repeat element isolated from a 2,4,5-T degrading strain of Pseudomonas cepacia. AB - Pseudomonas cepacia strain AC1100, capable of growth on 2,4,5 trichlorophenoxyacetic acid (2,4,5-T), was mutated to the 2,4,5-T- strain PT88 by a ColE1::Tn5 chromosomal insertion. Using cloned DNA from the region flanking the insertion, a 1477-bp sequence (designated RS1100) was identified which was repeated several times on the wild-type chromosome and was also present on AC1100 plasmid DNA. Various chromosomal fragments containing this sequence were cloned and their nucleotide sequence was determined. Examination of RS1100 revealed the presence of 38-39-bp terminal inverted repeats immediately flanked by 8-bp direct repeats. The translated sequence of the single large open reading frame of RS1100 showed structural similarity to the phage Mu transposase and other DNA-binding proteins. Thus the AC1100 repeated sequence has several structural features in common with insertion sequence elements. Three copies of RS1100 were mapped near 2,4,5-t genes encoding degradation of 5-chloro-1,2,4-trihydroxybenzene, an intermediate in 2,4,5-T degradation. Neither RS1100 nor the 2,4,5-t genes hybridized to DNA isolated from Pseudomonas strains, including P. cepacia, suggesting that both gene fragments may be of foreign origin recruited in strain AC1100. The origin of these two DNA segments as well as the role played by RS1100 in the recruitment of 2,4,5-t genes in AC1100 are presently under investigation. PMID- 2546861 TI - Nucleotide sequence of the Vibrio alginolyticus calcium-dependent, detergent resistant alkaline serine exoprotease A. AB - The nucleotide sequence of the Vibrio alginolyticus alkaline serine exoprotease A (ProA) gene cloned in Escherichia coli was determined. The exoprotease A gene (proA) consisted of 1602 bp which encoded a protein of 534 amino acids (aa) with an Mr of 55,900. The region upstream from the gene was characterized by a putative promoter consensus region (-10 -35), a ribosome-binding site and ATG start codon. The proA gene encodes a typical 21-aa N-terminal signal sequence which, when fused to alkaline phosphatase by means of transposon TnphoA, was able to mediate transport of the alkaline phosphatase to the periplasm in E. coli. Deletions of up to 106 aa from the C terminus of ProA did not result in the loss of extracellular protease activity. Additional V. alginolyticus genes were not involved in the secretion into the medium of the cloned ProA in E. coli. The amino acid sequence of ProA showed low overall homology to a Serratia marcescens serine exoprotease but significant homology was detected with other subtilisin family exoproteases. The fungal proteinase K, another sodium dodecyl sulfate resistant protease, had 44% aa homology with ProA. PMID- 2546862 TI - The HU protein is not essential for generating deletions adjacent to transposon Tn3. AB - Escherichia coli mutants deficient in the HU protein were used to test whether this protein is essential for generating DNA deletions adjacent to the ends of Tn3. There were no significant differences in the frequencies of the adjacent DNA deletions in the isogenic series which differ only in the HU loci (hupA or hupB), indicating that the HU protein is not essential for this reaction. PMID- 2546863 TI - The effect of oxygen, antioxidants, and superoxide radical on tyrosine phenoxyl radical dimerization. AB - Dimerization of tyrosine phenoxyl radical yields bityrosine (BT) which can easily be monitored by its characteristic fluorescence at 400 nm. The reactivity of tyrosine phenoxyl radical with O2 was examined by a variety of techniques. BT fluorescence was measured as a function of O2 concentration. Over a range of pH values (4-12) there was no effect of oxygen on BT production ([O2] less than or equal to 0.72 mM). In addition, oxygen uptake by the phenoxyl radical was measured directly with an oxygen electrode. It was determined by this technique that oxygen does not react with the phenoxyl radical with a rate constant greater than 10(3) M-1 s-1. Tyrosine phenoxyl radical "repair" by superoxide and physiological antioxidants was examined by BT fluorescence quenching as well as pulse radiolysis. Implications of these results as to the fate of tyrosine phenoxyl radicals produced in biological systems is discussed. PMID- 2546864 TI - The antioxidant action of N-acetylcysteine: its reaction with hydrogen peroxide, hydroxyl radical, superoxide, and hypochlorous acid. AB - N-acetylcysteine has been widely used as an antioxidant in vivo and in vitro. Its reaction with four oxidant species has therefore been examined. N-acetylcysteine is a powerful scavenger of hypochlorous acid (H--OCl); low concentrations are able to protect alpha 1-antiproteinase against inactivation by HOCl. N acetylcysteine also reacts with hydroxyl radical with a rate constant of 1.36 X 10(10) M-1s-1, as determined by pulse radiolysis. It also reacts slowly with H2O2, but no reaction of N-acetylcysteine with superoxide (O2-) could be detected within the limits of our assay procedures. PMID- 2546865 TI - Vanadate-stimulated oxidation of NAD(P)H. AB - Vanadate stimulates the oxidation of NAD(P)H by biological membranes because such membranes contain NAD(P)H oxidases which are capable of reducing dioxygen to O2- and because vanadate catalyzes the oxidation of NAD(P)H by O2-, by a free radical chain mechanism. Dihydropyridines, such as reduced nicotinamide mononucleotide (NMNH), which are not substrates for membrane-associated NAD(P)H oxidases, are not oxidized by membranes plus vanadate unless NAD(P)H is present to serve as a source of O2-. When [NMNH] greatly exceeds [NAD(P)H], in such reaction mixtures, one can observe the oxidation of many molecules of NMNH per NAD(P)H consumed. This reflects the chain length of the free radical chain mechanism. We have discussed the mechanism and significance of this process and have tried to clarify the pertinent but confusing literature. PMID- 2546866 TI - What do we measure by a luminol-dependent chemiluminescence of phagocytes? AB - The review presents a survey of published findings concerning the mechanism of luminol-dependent chemiluminescence in biological systems. The potential of various oxygen species (superoxide anion, hydrogen peroxide, hydroxyl radical) to react with luminol is discussed. The ability of commonly used enzymes (superoxide dismutase, catalase), inhibitors, and oxygen radical scavengers to discriminate between individual oxygen species is assessed together with the potential of a variety of substances encountered in biological systems to interfere in luminol dependent chemiluminescence reactions. It is concluded that luminol-dependent chemiluminescence gives at present very little ability to discriminate between individual oxygen or radical species. Furthermore, luminol-dependent chemiluminescence used in biological systems is extremely prone to many interferences, which are very difficult to control. PMID- 2546867 TI - Persistent free radicals in woodsmoke: an ESR spin trapping study. AB - Free radicals are detected in the gas-phase smoke resulting from the combustion of wood using the electron spin resonance (ESR) spin trapping method. The materials were pyrolyzed by rapid heating in a quartz tube in a flowing air stream. The filtered smoke was bubbled into a dodecane solution of alpha-phenyl-N tert-butyl nitrone, and the resulting nitroxide radicals were detected by ESR. The radicals spin trapped from woodsmoke are compared to those we have spin trapped from tobacco smoke; the smoke from both yellow pine and oak produce more intense ESR spectra than does tobacco smoke per unit mass burned under the conditions of these experiments. When woodsmoke is bubbled through pure dodecane and the resulting woodsmoke/dodecane solution is held for a delay time before the PBN is added, radicals are detected even after the woodsmoke/dodecane solution is aged for more than 20 min. Similar experiments with tobacco smoke show that radicals no longer are trapped even after much shorter delay times from tobacco smoke/dodecane solutions. PMID- 2546868 TI - Relationship between superoxide anion radical generation and aging in the housefly, Musca domestica. AB - The objective of this study was to elucidate the possible cause of increased oxidative stress observed in the adult housefly during aging. The hypothesis that increased production of oxygen radicals may be a cause of the increased oxidative stress was tested by comparison of 8-day and 15-day old flies, which represent the stage of full maturation and the beginning of the dying phase, respectively. Rates of both antimycin A-resistant respiration of isolated mitochondria and O2 generation at ubiquinone-cytochrome b site by submitochondrial particles increased during aging and were associated with life expectancy of flies. Flies destined to die earlier than their cohorts of the same age exhibited a relatively higher rate of O2- production. Age-related increase in O2- generation was not associated with corresponding changes in ubiquinone content of mitochondria. PMID- 2546869 TI - The action of hydrogen peroxide on paired pulse and long-term potentiation in the hippocampus. AB - The action of a reactive oxygen intermediate, that is, hydrogen peroxide (H2O2) on modulation of synaptic transmission was examined in the hippocampal brain slice preparation. Microinjection of H2O2 into the apical dendritic region of the CA1 pyramidal cells produced no change in either the pattern or amplitude of paired pulse facilitation compared to saline injection (control). Long term potentiation (LTP), induced by high frequency stimulation of homosynaptic inputs, however, was blocked by microinjection of H2O2 into the dendritic tree. LTP was seen in only 2 out of 10 slices investigated when treated with H2O2 while LTP was seen in 4 out of 5 slices when saline injected. The results suggest that a reactive oxygen intermediate can selectively modify synaptic mechanisms in the hippocampus. PMID- 2546870 TI - Evaluation of dibromonitrosobenzene sulfonate as a spin trap in biological systems. AB - In the present study dibromonitrosobenzene sulfonate (DBNBS) was examined for its suitability for spin trapping for ESR detection of superoxide radicals in biological systems. This nitroso spin trap recently has been reported to yield very persistent spin adducts with O2. as well as with various carbon-centered radicals. In the present work the possible toxicity of DBNBS, the partitioning of its spin adducts into cells, and the stability of the adducts and the parent compound inside cells were studied. No significant toxicity was found. In cellular systems, however, DBNBS did not produce detectable adducts with O2.; it also did not detectably trap superoxide generated in the xanthine/xanthine oxidase system. Both DBNBS and a DBNBS adduct performed extracellularly and then added to cell suspensions were rapidly metabolized by cells. Intracellular spin adducts were not detected under any condition. Evidently, in spite of its promising features, DBNBS will not be useful for spin trapping radicals in cellular systems or for detecting superoxide radicals in any biological system. PMID- 2546871 TI - Does dietary fibre stimulate intestinal epithelial cell proliferation in germ free rats? AB - The aim of the present experiment was to investigate the role of hind gut fermentation in the proliferative response of the intestinal epithelium to dietary fibre. We have previously shown that refeeding starved rats with an elemental diet supplemented with fermentable dietary fibre (but not inert bulk) is capable of stimulating intestinal epithelial cell proliferation throughout the gastrointestinal tract. Three groups of 10 germ free (GF) rats and three groups of 10 conventional (CV) rats, were used. All groups were starved for three days and then refed for two days with either an elemental diet (Flexical); Flexical plus 30% kaolin; or Flexical plus 30% of a fibre mixture. Cell production was determined by the accumulation of vincristine arrested metaphases in microdissected crypts. There was no significant difference between refeeding the rats with an elemental diet alone or with kaolin supplementation, however, the addition of fibre in CV rats was associated with a significant increase in intestinal crypt cell production rate in both the small intestine (p less than 0.01) and the colon (p less than 0.001). This marked proliferative effects of fibre was abolished in the GF rats. It can be concluded that it is the products of hind gut fermentation, not fibre per se that stimulate intestinal epithelial cell proliferation in the colon and small intestine. PMID- 2546872 TI - Hepatocellular carcinoma in idiopathic haemochromatosis. PMID- 2546873 TI - Primary mucinous adenocarcinoma developing in an ileostomy stoma. PMID- 2546875 TI - Eclampsia as a possible risk factor for persistent trophoblastic disease. AB - Twenty cases of hydatidiform mole complicated by eclampsia with adequate postevacuation follow-up are identified in a review of the literature since 1866. The clinical presentation of each of these patients is reviewed with particular attention to the existence of known risk factors for persistent trophoblastic disease. After excluding 3 women with coexisting fetus and 2 others who were treated initially with a total abdominal hysterectomy, it was discovered that 14 of the remaining 15 women developed persistent trophoblastic disease. This frequency of persistent trophoblastic disease is greater than can be explained based on previously described risk factors and suggests that the occurrence of eclampsia may be an independent risk factor for persistent trophoblastic disease. PMID- 2546874 TI - Adenoid cystic carcinoma of Bartholin's gland: a report of five new cases treated with surgery and radiotherapy. AB - Adenoid cystic carcinoma (ACC) is a slow-growing tumor with a marked tendency for perineural and local invasion. The neoplasm occurs more frequently in the head and neck region and only 39 previously described cases of adenoid cystic carcinoma of Bartholin's gland are found in the world literature. Most authors advocate surgery as the primary treatment. Several cytostatic drugs have been tried but the results have been poor. The benefit of radiotherapy in the treatment of this type of tumor is not yet established. This is a report of five new cases all treated with surgery and postoperative radiotherapy. Three patients had histopathologically proven evidence of residual disease after surgery and were subsequently treated with radiotherapy. They are now living without evidence of disease after 28, 51, and 138 months. PMID- 2546876 TI - Fructose-bisphosphatase-deficient mutants of mucoid Pseudomonas aeruginosa. AB - Fructose-bisphosphatase-deficient mutants of mucoid Pseudomonas aeruginosa were isolated by ethyl methanesulfonate mutagenesis using gluconate as the nonpermissive substrate, and all the sixty isolates possessed 10-30% of the parental enzyme activity. The mutants had low levels of fructose-biphosphate aldolase activity and could not normally synthesize alginate from any substrate except on Pseudomonas isolation agar plates. The results suggest the essentiality of fructose bisphosphatase activity for the growth or survival of P. aeruginosa and a probable linkage of genes controlling this enzyme with those of fructose bisphosphate aldolase and alginate biosynthesis. PMID- 2546877 TI - [Can HIV infection be competitively inhibited? New therapeutic approaches- development of an immuno-adhesin]. PMID- 2546878 TI - Possible binding sites for inositol 1,4,5-trisphosphate in canine tracheal smooth muscle cells and rat liver cells. AB - To search for binding site of inositol 1,4,5-trisphosphate (InsP3) in relation to the release of Ca2+ from smooth muscle cells of the canine trachea, InsP3 coupled with p-azidobenzoyl beta-alanine (AB beta A) was synthesized for photoaffinity labelling, using N,N'-carbonyldiimidazole. With subsequent photolysis of the smooth muscle cells following incubation with this derivative (InsP3-AB beta A), the InsP3-induced Ca2+ release from saponin-permeabilized dispersed smooth muscle cells no longer occurred. The irreversible release of Ca2+ induced by InsP3-AB beta A was prevented by adding 10-fold excess amounts of InsP3. These observations strongly suggest that there is a covalent binding of InsP3-AB beta A to a specific "receptor" on the sarcoplasmic reticulum in tracheal smooth muscle cells. To determine the specific binding sites on the endoplasmic reticulum, I synthesized an isotope-labelled arylazide derivative of InsP3; InsP3 was coupled with p-[125I]azidosalicyl beta-alanine, using carbonyldiimidazole as a catalyst. I obtained three proteins with molecular weights of 44K-Da, 27K-Da and 22K-Da, which were preferentially labelled with the [125I]arylazide derivative of InsP3 (InsP3-[125I]AS beta A) in photoirradiated microsomal fractions of canine tracheal smooth muscle cells. The labelling of these proteins was partially blocked by an excess amount of unlabelled InsP3 but not by inositol 1,4 bisphosphate (Ins(1,4)P2). Similar binding proteins with molecular weights of 48K Da, 32K-Da and 18K-Da were also labelled in the endoplasmic reticulum fraction of the rat liver, but such proteins were not evidenced in the plasma membrane fraction. PMID- 2546879 TI - Spontaneous regression of a hepatocellular carcinoma--a case report. AB - Spontaneous regression of hepatocellular carcinoma was observed in a 67-year-old Japanese male with liver cirrhosis. Although the patient showed no histological evidence of hepatocellular carcinoma, findings obtained by hepatic angiography, computed tomography and echography strongly indicated that the tumor was a hepatocellular carcinoma. Moreover, the rate of growth of the tumor nodule paralleled an increase in serum alpha-fetoprotein. Following this accelerated increase in serum alpha-fetoprotein a sudden drop was observed with no specific treatment for cancer, and regression of the tumor was also observed. Six months later the serum alpha-fetoprotein level had normalized, and the tumor had disappeared completely. The patients is now alive and doing well more than 70 months after the initial diagnosis of hepatocellular carcinoma. PMID- 2546880 TI - Recurrent form of hepatocellular carcinoma after partial hepatic resection. AB - An analysis of 27 recurrent cases after partial hepatic resection for hepatocellular carcinoma, closely followed-up with various diagnostic imagings, is reported. The patients comprised 22 males and 5 females. Their ages ranged from 34 to 76 years, with an average of 57.8 years. Twenty-five had underlying cirrhosis and 2 presented chronic hepatitis histologically. The 27 patients were classified into the following three types. Type I comprised 18 patients with local recurrence. Local recurrence means that the tumor recurred in an adjacent segment to that initially affected or in the same segment. Thirteen of these patients had a recurrence within a year-after the initial operation. Type II comprised 6 cases of multiple intrahepatic metastases, and Type III 3 cases which were thought to represent a metachronous occurrence. Five of the 6 cases of Type II had multiple intrahepatic metastases within 3 months after the initial operation. The results of the present series indicate that when patients with hepatocellular carcinoma are operated on, manipulation should be as careful as possible in order not to disperse the cancer cells, and close follow-up with imagings more than once every three months, especially in the first year after the initial operation should be ensured wherever possible. PMID- 2546882 TI - Human papillomavirus and cervical cancer. PMID- 2546883 TI - Human papillomavirus and cervical cancer. Summary report. PMID- 2546881 TI - Thyroid microsomal/thyroid peroxidase autoantibodies show discrete patterns of cross-reactivity to myeloperoxidase, lactoperoxidase and horseradish peroxidase. AB - The recent cloning of the thyroid peroxidase (TPO) has shown that it is identical to the thyroid microsomal antigen (TMA), a potent antigen involved in autoimmune thyroid disease (ATD), which shares significant sequence homology with myeloperoxidase. The present study shows that autoantibodies (aAb) to the TMA/TPO antigen cross-react with human leucocyte myeloperoxidase, bovine lactoperoxidase and horseradish peroxidase. Cross-reactivity to myeloperoxidase was only apparent by ELISA using reduced and alkylated antigen preparations or by immunoblotting following denaturation with SDS. Sequential absorption of sera on SDS-denatured thyroid microsomes immobilized on Sepharose-4B followed by absorption on native microsomes removed all aAb specificities to TMA/TPO and the three peroxidase preparations, giving compelling evidence on the genuine cross-reactive nature of these aAbs. Sera from different patients contain different qualitative and quantitative specificities of aAb to the TMA/TPO antigen, confirming the polyclonal nature of this autoimmune response. PMID- 2546884 TI - Experimental evidence on oncogenicity of papillomaviruses. PMID- 2546885 TI - Incidence, prevalence and time trends of genital HPV infection determined by clinical examination and cytology. PMID- 2546887 TI - Human papillomavirus and cervical neoplasia: a critical review of the epidemiological evidence. PMID- 2546886 TI - Measurement error in epidemiological studies of human papillomavirus and cancer. PMID- 2546888 TI - Human cervical papillomavirus infection: a clinical perspective. PMID- 2546889 TI - Human papillomavirus-related changes in the genital tract. PMID- 2546890 TI - Diagnosis of HPV by DNA hybridization techniques. PMID- 2546891 TI - Activation of neutrophil collagenase by cathepsin G. AB - Collagenase is secreted from neutrophils as a latent or proenzyme. In an effort to understand the mechanism of collagenase activation in inflammation, human peripheral neutrophils (PMNs) were isolated and incubated with the tumor promotor, phorbol myristate acetate (PMA), which induces the neutrophils to degranulate and secrete proteinases. Neutrophil media were then treated with various activators or inhibitors of collagenase and other proteinases, and the collagenase activity was measured. A serine proteinase secreted from neutrophils, cathepsin G, was found to activate latent collagenase, but it was also found to require activation itself. Both hypochlorous acid (HOCl) and oxidized glutathione (GSSG) were tested for their collagenase-activating ability and were found to be successful only in the presence of active cathepsin G. A specific cathepsin G inhibitor (0.5 mM Z-Gly-Leu-Phe-CH2Cl) prevented the activation of latent collagenase by HOCl. To confirm these results, purified neutrophil cathepsin G was incubated with a neutrophil proteinase mixture which contained latent collagenase. The collagenase was shown to be activated upon incubation with purified cathepsin G. These results indicate that cathepsin G is a key mediator in neutrophil collagenase activation. PMID- 2546892 TI - Macrophage-produced oxygen radical generating activities for polymorphonuclear leukocytes. AB - Guinea pig peritoneal macrophages were stimulated in vitro by bacterial lipopolysaccharide. After incubation, the supernatants of macrophage cultures were collected and tested for O2- production on guinea pig peritoneal polymorphonuclear leukocytes. The supernatants of macrophage cultures stimulated by lipopolysaccharide had significantly higher levels of O2- -generating activities in polymorphonuclear leukocytes, and these activities appeared in the macrophage cultures within 2 h after stimulation by lipopolysaccharide. However, the supernatants obtained from the nonstimulated cultures could not produce these activities. These activities disappeared with heating or trypsin and were not produced in macrophage cultures by incubation with cycloheximide. PMID- 2546893 TI - Permeabilization and calcium-dependent activation of rabbit polymorphonuclear leukocytes by poly-L-arginine. AB - In the absence of extracellular Ca2+, poly-L-arginine induces little lysozyme release from rabbit polymorphonuclear leukocytes (PMNs). The polycation causes plasma membrane damage, which is evident from the release of the cytoplasmic enzyme lactate dehydrogenase (LDH). In the presence of Ca2+ concentrations higher than 0.2 mM, poly-L-arginine induces a strong lysozyme release that is superimposed on the membrane-damaging effect. The results suggest that poly-L arginine permeabilizes the plasma membrane, enabling Ca2+ to enter the cell, which results in the exocytotic release of granule constituents. The GTP analog GTP gamma S shifts the Ca2+ requirement of exocytosis to slightly higher concentrations, whereas it completely inhibits poly-L-arginine-induced LDH release. Pertussis toxin gives a moderate inhibition, and La3+ completely inhibits poly-L-arginine-induced enzyme release. Whereas poly-L-arginine alone induces little superoxide generation in rabbit PMNs, there is a synergistic enhancement of superoxide production when GTP gamma S and poly-L-arginine are present together. Guanine nucleotides apparently have a modulating effect on the actions of poly-L-arginine on the PMN, but the nature of this effect remains to be determined. PMID- 2546894 TI - Modulation of functional activity of human polymorphonuclear and mononuclear phagocytes by intravenous gamma globulin. AB - Intravenous gamma-globulin was tested in a range of concentrations compatible with the increments obtained after therapeutic infusions for modulation of phagocytic functions of human polymorphonuclears (PMNs) and monocytes. Intravenous gammaglobulin in concentrations of 3.0 mg/ml or more increased adhesiveness and suppressed chemotaxis of PMNs. There was marked dose-dependent enhancement of opsonization of gram-positive and gram-negative microorganisms. Preincubation of PMNs with intravenous gamma-globulin caused enhancement of the total bacteria ingested, total bacteria killed, phagocytosis, and phagocytic index, when gram-positive and gram-negative bacteria were tested. During phagocytosis, there was no release of LDH or lysozyme; however, there was release of beta-glucuronidase. No significant difference in phagocytic enhancement was found when filtered and native intravenous gamma-globulin preparations were compared. There was marked enhancement of the superoxide anion generation by intravenous gamma-globulin above the concentration of 0.01 mg/ml. Intravenous gamma-globulin also markedly enhanced phagocytic activity of monocytes. Therefore, intravenous gamma-globulin modulates not only opsonization-related phenomena, but also exerts a complex influence on other aspects of phagocytic activity. PMID- 2546895 TI - Etiology of childhood hypothyroidism. AB - This study prospective without any selection bias included 80 of the 152 hypothyroid infants and children seen over the past six years. The clinical diagnosis was confirmed by TSH and thyroid hormone (T3, T4) studies. Scanning for thyroid with TC99m pertechnetate was carried out in all except seven older children with grade II and III goiters where 131I uptake studies were done. Serum thyroglobulin (RIA) was estimated and antithyroglobulin and antimicrosomal antibodies were tested. Based on thyroid 131I scan or 131I uptake, 52.5% had no demonstrable thyroid tissue except one with hypoplasia (Group I, n = 42), 25% had ectopic thyroid (Group II, n = 20), and 22.5% had normal or enlarged thyroid gland (Group III, n = 18). One hypothyroid patient of Group III had thyroiditis with high antibody titre and one was proved to have iodine deficiency). The mean age at time of diagnosis was lowest in Group I (age in months--30.3 +/- 36.2; 60.6 +/- 53.9; 106.2 +/- 69.3 in Groups I, II and III respectively. The intergroup differences in age were significant. The mean serum Tg levels increased progressively from Groups I to III. In the present series thyroid dysgenesis led to hypothyroidism in 77.5%, with athyreosis in 52.5% and ectopia in 25%. Dyshormonogenesis was noted in 20% and thyroiditis in 1.5%. PMID- 2546896 TI - Essential fatty acids and eicosanoids in cutaneous inflammation. PMID- 2546897 TI - Leishmaniasis. A worldwide problem. PMID- 2546898 TI - Noncontrast and contrast enhanced MR imaging in the evaluation of partial ureteral obstruction: an experimental study in the micropig. AB - Twelve Yucatan micropigs (3 controls; 3 sham-operated; 6 with unilateral obstruction) were studied to assess the value of noncontrast and contrast enhanced (Gadolinium-DTPA) magnetic resonance (MR) imaging in the evaluation of partial ureteral obstruction. MR findings were correlated with findings of quantitative (Tc-99m-DMSA) scintigraphy, and histology. On noncontrast T1 weighted images, the normal porcine kidney demonstrated good corticomedullary contrast (CMC = 16.8% +/- 5.0). Five minutes after administration of Gd-DTPA, there was enhancement of the renal cortex (+24.4% and medulla (+46.2%), and CMC was no longer discernible. Enhancement of the urine within the collecting system (+119.1%) was also observed. The obstructed kidneys demonstrated marked thinning of the renal parenchyma and decreased signal intensity on noncontrast T1- and T2 weighted images (P less than 0.01). Urine in the dilated collecting system did not differ significantly from urine in controls except in the three animals with urinary tract infection (P less than 0.05). Five minutes following injection of Gd-DTPA, there was enhancement of the renal parenchyma in all kidneys. Excretion was seen in three pigs and no excretion in two. Thus, useful information can be obtained in partial ureteral obstruction from both pre-contrast and Gd-DTPA enhanced MR images of the kidney. PMID- 2546899 TI - Characterization of DNA polymerases in an uninfected and virus PBCV-1-infected green alga--Chlorella strain NC64A. AB - Chlorella NC64A cells infected with the large double-stranded DNA-containing virus PBCV-1 and uninfected cells were assayed for DNA polymerase activity. Both uninfected and infected cells contained three forms of DNA polymerase activity: (i) an exogenous DNA-dependent 10,000 g soluble fraction, (ii) an exogenous DNA dependent 10,000 g particulate fraction, and (iii) a DNA-independent 10,000 g particulate fraction. The three DNA polymerase activities in the infected and uninfected cells were distinguished from one another by the conditions required for optimum activity and by their sensitivity to inhibitors. PMID- 2546900 TI - Inhibition of chemical Epstein-Barr virus induction by dimethyl sulfoxide and related polar compounds. AB - Dimethyl sulfoxide and twelve related polar compounds, capable of inducing differentiation of murine erythroleukemia cells, markedly inhibited induction of Epstein-Barr virus antigens in virus producer and nonproducer lymphoblastoid cells exposed to 12-O-tetradecanoylphorbol-13-acetate, n-butyric acid, 5-iodo-2' deoxyuridine, and a combination of 12-O-tetradecanoylphorbol-13-acetate and n butyric acid. The concentration at which these compounds caused 90% inhibition of antigen induction corresponded to that optimal for induction of differentiation. At the same concentrations, polar compounds did not affect cell viability or virus induction by superinfection with Epstein-Barr virus recovered from P3HR-1 cells. The inhibitory activity was reversible. Three nonpolar differentiating compounds revealed no inhibitory activity. PMID- 2546901 TI - [The effect of infrared laser rays on herpes simplex virus and the functions of human immunocompetent cells]. AB - In vitro investigations were performed to study the effect of infrared Nd:YAG laser irradiation on herpes simplex virus and the viral replication in herpes infected Vero cell microcultures. In addition, the influence of laser irradiation on human immuncompetent cells was investigated by irradiation and incubation of herpes-infected cell cultures overlaid with leucocytes and observation of leucocyte migration under agarose after laser irradiation. There was no evidence of herpes simplex virus inactivation by laser irradiation. Irradiation of Vero cell microcultures infected with virus did not influence the development of the viral cytopathic effect. However, irradiated cultures showed an increase of about 50% in the virus yield. Only a slight indication of laser influence on immunocompetent cells was found. Cell cultures incubated with leucocytes (from both seropositive and seronegative donors) and irradiated showed an inhibitory effect with about half the virus yield seen in unirradiated controls. However, there was no relation between the energy applied and yield reduction. In addition, laser light caused no change in leucocyte migration. PMID- 2546902 TI - [Breast cancer before 36 years of age]. AB - Breast cancer in young women is a dreaded disease of bad prognosis, classically worse than for older women. A local study was undertaken in Lausanne to evaluate this notion. 94 cases aged less than 36 years were collected over 15 years, the incidence being 9 new cases per years per 100,000 under-36 women. 76% of the tumors were discovered accidentally by the patient, 24% by a physician during a routine breast examination. Delays from first symptom to histologic diagnosis were short, averaging 19 weeks. 80 mammographies were performed on the 94 cases, of whom 50 positives for a cancer, and 29 negative or doubtful. 22 of these negative 29 were reevaluated, 7 being positive for a tumor and 15 negative, showing either a benign mass or no lesion at all. 6 of the 7 positives were misdiagnosed in easy-to-read breasts, the false interpretation being chiefly caused by the young age of the patients. Pathologically, the rate of invasive ductal carcinoma was very high at 91.4%, which is the highest in the literature, and much higher than for older women in Lausanne (68%). Survival was 84% at two years, 63% at five years and 46% at 10 years, all deaths due to breast cancer. Those rates are better, globally and stage by stage, than for older women or than other reports on young women. We conclude that breast cancer in young women has a better prognosis than formerly thought. PMID- 2546903 TI - Nephrotoxicity of pyrroloquinoline quinone in rats. AB - When pyrroloquinoline quinone (PQQ) was intraperitoneally injected into rats daily for 4 days at a dose of 11.5 mg/kg body weight/injection, functional and morphologic changes of the kidneys were clearly observed. The most prominent finding was necrotic and degenerative changes of the proximal tubular epithelium as well as hematuria and an elevation of serum creatinine concentration. PMID- 2546904 TI - Nonfunctioning islet cell pancreatic cancer. AB - Nonfunctioning islet cell tumors are rarer than functioning neuroendocrine pancreatic tumors. Although they share certain characteristics with functioning tumors, they do not show clinical evidence of hormonal secretion. Their symptoms usually arise as a result of the effect of the mass, jaundice, weight loss, and malaise. They are more likely to be malignant than functioning tumors. They are slow growing, however, and their size and malignancy do not preclude long survival. For these reasons they warrant aggressive surgical intervention. PMID- 2546905 TI - Bilateral breast carcinoma treated with definitive irradiation. AB - From 1977 to 1987, 30 women were treated with definitive irradiation following breast-conserving surgery for bilateral carcinoma of the breast for a total of 60 treated breasts. Eleven women presented with concurrent bilateral carcinoma, and 19 women had sequential bilateral carcinoma. Pathologic axillary staging was performed in 51 of the 60 treated breasts. A total dose of greater than or equal to 6,000 cGy was delivered from breast tangential irradiation plus an electron or Iridium boost to 95% (57/60) of the treated breasts. A third field was used to treat the regional axillary and supraclavicular lymph nodes bilaterally in three women (10%) and unilaterally in ten women (33%). Tangential fields were matched at midline in 17 patients, and in ten patients, the tangential fields overlapped by up to 3 cm on skin. In two patients, the tangential fields were matched to an internal mammary nodal field, and in one patient, tangential fields were matched to a mediastinal field given for postoperative radiotherapy for lung cancer. For the overall group of 30 patients, the 5-year actuarial NED survival following treatment of the first breast cancer was 79%, and the 5-year actuarial relapse free survival was 72%. For the 60 treated breasts, the 5-year actuarial local failure rate was 6%. An analysis of complications and cosmesis showed results similar to previously reported results for unilateral breast cancer. These results show that definitive irradiation following breast-conserving surgery for patients with bilateral breast cancer can technically be delivered with low complication rates and with acceptable survival and local control rates. Definitive irradiation should be considered as an acceptable alternative treatment to bilateral mastectomy for appropriately selected patients with concurrent or sequential bilateral early stage carcinoma of the breast. PMID- 2546906 TI - Importance of radiation dose in achieving improved loco-regional tumor control in limited stage small-cell lung carcinoma: an update. AB - Between 1974 and 1986, 576 patients (284 limited and 292 extensive stages) were treated at this institution. To keep multiagent chemotherapy (CT) at a uniform intensity, patients who received (a) combined modality approach of both multiagent chemotherapy and thoracic radiotherapy (RT) and (b) greater than or equal to 3 cycles of multiagent chemotherapy (greater than or equal to 3 drugs), were chosen for this analysis. Out of 284 patients with limited Stage small-cell lung carcinoma, there were 154 such patients who met these strict criteria, and the treatment methods for the remaining 130 patients were as follows: (a) chemotherapy alone with radiotherapy reserved for local failure (47 pts); (b) radiotherapy alone (20 pts); (c) surgery +/- adjuvant chemotherapy or radiotherapy (37 pts); (d) modified chemotherapy plus radiotherapy (26 pts). During the 12-year period, the therapeutic factors have evolved. Radiation-dose was increased from 30-40 Gy (time dose fractionation 49-66) in 1974-1977 to 44-52 Gy (time dose fractionation 73-86) in 1978-1986. The target volume for radiotherapy included the primary lesion with a 2-cm margin of normal lung and the mediastinum. Chemotherapy program also evolved from COP, CAV (1974-1977) to MACC, VCE-VCA, PCE-ACE (1978-1986). Fifty of 154 patients (32%) developed loco regional recurrence (infield failure) and 98% (49/50) of these patients exhibited this by 2.5 years. Survival data of 154 patients were as follows: (a) Median survival time (MST) was 12 M; (b) actuarial survival rates at 2 and 5 years were 21% and 8%, respectively. Fifty percent of these patients died within 12 months (MST 12 M) and were not exposed to the full length of the risk period for loco regional failure. To take into account the duration of exposure to the risk period, actuarial method was employed to measure the probability of loco-regional failure. Loco-regional failure rates at 2.5 years were 37%, 39%, 49%, 79%, and 84% for 50 Gy, 45 Gy, 40 Gy, 35 Gy, and 30 Gy, respectively. The difference between the recurrence rates of 37% and 79% by 50 Gy and 35 Gy was statistically significant, p less than 0.05. Although the recurrence rates of 37% and 49% by 50 Gy and 40 Gy were not statistically different, there was a strong trend of a better control rate of loco-regional carcinoma by higher radiation doses. The time to recurrence seems also shorter with lower radiation-dose than that of higher radiation doses.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2546907 TI - Lymphatic vessel invasion is an independent prognostic factor for survival in colorectal cancer. AB - To assess whether lymphatic vessel invasion (LVI) is an independent prognostic factor in colorectal cancer, we retrospectively reviewed the records of 462 patients who underwent potentially curative surgery for carcinoma of the colon and rectosigmoid/rectum (rs/rectum) at the New England Deaconess Hospital from 1965-1978. Sixty-one patients were identified as having tumors with lymphatic vessel invasion (LVI+), and they were compared with the remaining group of 401 patients who had tumors without lymphatic vessel invasion (LVI-). The incidence of lymphatic vessel invasion was significantly increased in tumors with blood vessel invasion (24% vs. 5%, p = 0.000001). Patients with LVI+ tumors also had a significantly increased incidence of positive nodes (59% vs. 25%, p = 0.0004), the average number of positive nodes (4.8 vs. 2.2, p = 0.0003), and a lower 5 year survival rate (colon: 57% vs. 84%, p = 0.0001; rs/rectum: 38% vs. 71%, p = 0.004). There was a significant (p less than or equal to 0.05) increase in local (16% vs. 7%), abdominal (33% vs. 9%), and distant (13% vs. 4%) failure as a component of component of failure in patients with LVI+ colon cancer and a significant increase in abdominal (33% vs. 11%) and distant (13% vs. 8%) failure as a component of failure in patients with LVI+ rectosigmoid/rectal cancer. Proportional hazards analysis demonstrated that lymphatic vessel invasion was an independent prognostic factor for survival. PMID- 2546908 TI - Laryngeal paralysis in immature and mature dogs as one sign of a more diffuse polyneuropathy. AB - Six dogs with laryngeal paralysis had clinical, electrophysiologic, and pathologic evidence of a more generalized polyneuropathy. Three of the dogs were young Dalmatians, one was a young Bouvier des Flandres, and two were older, large breed dogs. The results of this study suggest that laryngeal paralysis in dogs may frequently be one clinical sign of an underlying, more generalized polyneuropathy. Two forms of this generalized polyneuropathy may exist: an early form, as seen in young dogs with congenital or hereditary disease, and a delayed onset form that is usually found in older dogs with so-called idiopathic laryngeal paralysis, some of which may have hypothyroidism. PMID- 2546909 TI - Selective killing of transformed fibroblasts by combined treatment with cycloheximide and aphidicolin. AB - The possibility of selective killing of transformed cells in a mixed population of untransformed and transformed cells was examined using a cell culture system of rat 3Y1 fibroblasts (parental 3Y1 cells, 3Y1 cells transformed with either SV40, polyoma virus, Rous avian sarcoma virus, E1A gene of adenovirus type 12, or H-v-ras oncogene). The principle of the selective killing is as follows. Under suboptimal culture conditions, untransformed cells are inhibited from progressing through G1 phase and retain viability, while transformed cells are not arrested. When DNA synthesis is inhibited for a long period, both types of cells in S phase die. Therefore, if we administer inhibitors of G1 progression and of DNA synthesis simultaneously to a cell population consisting of untransformed and transformed cells, most untransformed cells are arrested in G1 phase, retaining viability, while transformed cells leak from the G1 phase, cease DNA synthesis, and gradually die The present study shows that all types of transformants in stationary-phase cultures (consisting of cells mainly with a G1 DNA content) were killed to higher extents compared with untransformed cells, during incubation at lower cell densities with a combination of cycloheximide (G1 inhibitor) and aphidicolin (DNA-synthesis inhibitor). However, cycloheximide reduced the killing effect of aphidicolin by changing the irreversible DNA-synthesis inhibition to a reversible inhibition. The availability of G1 inhibitors that do not interfere with the irreversibility of inhibition of DNA synthesis is required for the treatment of cancer based on this idea. PMID- 2546910 TI - KS-501 and KS-502, new inhibitors of Ca2+ and calmodulin-dependent cyclic nucleotide phosphodiesterase from Sporothrix sp. AB - New inhibitors of Ca2+ and calmodulin-dependent cyclic-nucleotide phosphodiesterase, KS-501 and KS-502 were isolated from a fungus, Sporothrix sp. KAC-1985. Inhibitory concentration causing 50% inhibition (IC50) values of KS-501 and KS-502 for bovine brain enzyme were 1.8 and 4.3 microM, respectively. The compounds exhibited no or weak inhibition for calmodulin-independent cyclic nucleotide phosphodiesterases and protein kinase C. PMID- 2546911 TI - Inhibitory effect of antibiotic cerulenin on the respiratory burst in phagocytes. II. Inhibition by cerulenin of intracellular calcium mobilization in human neutrophils. AB - Generation of superoxide anion (O2-) and mobilization of intracellular Ca2+ in human neutrophils upon exposure to stimuli such as N formylmethionylleucylphenylalanine (fMLP) were inhibited by the antibiotic cerulenin, which inhibits fatty acid and cholesterol biosynthesis. The inhibition of O2- generation and Ca2+-mobilization required certain periods of incubation with cerulenin and both abilities of the cells were gradually lost following a similar time-course. In contrast, significant Ca2+-influx from the medium was observed in cerulenin-treated cells as well as untreated cells. The results suggest that an event which coincides with the Ca2+-mobilization and not Ca2+ per se is important for the induction of O2- generation in the fMLP-stimulated cells and that this step is blocked in cerulenin-treated cells. Phorbol myristate acetate or synthetic 1-oleoyl-2-acetylglycerol were able to bypass the block and induced O2- generation in cerulenin-treated cells. PMID- 2546912 TI - Bacterial overgrowth in the jejunum of ICR mice and Wistar rats orally administered with a single lethal dose of fusarenon-X, a trichothecene mycotoxin. AB - A single oral dose of fusarenon-X (F-X), a trichothecene mycotoxin, resulted in abnormal microflora in the jejunum in ICR mice and Wistar rats with some differences in dose response between the species. In the acute phase, enterobacteria, streptococci, Clostridium perfringens and bacteroides showed remarkably increased counts in the jejunum of mice and rats dosed with F-X while lactobacilli showed a decrease in count. F-X brought an invasion of Pseudomonas aeruginosa into the livers, lungs, kidneys and spleens of ICR mice. Changes in the jejunal microflora appeared after 7 h in ICR mice and after 24 h in Wistar rats after a single oral dose of F-X of 7.5 and 4.0 mg/kg b.w., respectively, and the microflora returned to its normal state at 72 h in mice and 96 h in rats. The changes of intestinal microflora were followed by alterations in the growth curves of both animal species. The pH in the glandular stomach was also greatly enhanced before changes in the jejunal microflora. Acute F-X intoxication may be an involved manifestation of essential cytotoxicity of F-X mycotoxin alone and secondary bacterial overgrowth in the bowel. PMID- 2546914 TI - Purification and characterization of S-adenosyl-L-methionine: uroporphyrinogen III methyltransferase from Pseudomonas denitrificans. AB - S-Adenosyl-L-methionine:uroporphyrinogen III methyltransferase (SUMT), the enzyme of the cobalamin biosynthetic pathway which catalyzes C methylation of uroporphyrinogen III, was purified about 150-fold to homogeneity from extracts of a recombinant strain of Pseudomonas denitrificans derived from a cobalamin overproducing strain by ammonium sulfate fractionation, anion-exchange chromatography, and hydroxyapatite chromatography. The purified protein has an isoelectric point of 6.4 and molecular weights of 56,500 as estimated by gel filtration and 30,000 as estimated by gel electrophoresis under denaturing conditions, suggesting that the active enzyme is a homodimer. It does not contain a chromophoric prosthetic group and does not seem to require metal ions or cofactors for activity. SUMT catalyzes the two successive C-2 and C-7 methylation reactions involved in the conversion of uroporphyrinogen III to precorrin-2 via the intermediate formation of precorrin-1. In vitro studies suggest that the intermediate monomethylated product (precorrin-1) is released from the protein and then added back to the enzyme for the second C-methylation reaction. The pH optimum was 7.7, the Km values for S-adenosyl-L-methionine and uroporphyrinogen III were 6.3 and 1.0 microM, respectively, and the turnover number was 38 h-1. The enzyme activity was shown to be completely insensitive to feedback inhibition by cobalamin and corrinoid intermediates tested at physiological concentration. At uroporphyrinogen III concentrations above 2 microM, SUMT exhibited a substrate inhibition phenomenon. It is suggested that this property might play a regulatory role in cobalamin biosynthesis in the cobalamin-overproducing strain studied. PMID- 2546913 TI - Implication of nifA in regulation of genes located on a Rhizobium meliloti cryptic plasmid that affect nodulation efficiency. AB - We examined the contribution of a cryptic plasmid, pRmeGR4b, to the nodulation of Medicago sativa by strain GR4 of Rhizobium meliloti. A 905-base-pair PstI DNA fragment in pRmeGR4b was found to hybridize DNA of the R. meliloti fixA promoter region as a probe. Sequence analysis of the PstI fragment showed a 206-base-pair region displaying high homology with the DNA upstream of the RNA start points of the P1 and P2 symbiotic promoters. Putative nif promoter consensus sequences were conserved in this DNA segment. Expression of DNA downstream of the nif promoterlike sequence, monitored by beta-galactosidase activity of different lacZ fusions, was demonstrated to depend on a functional nifA gene, both in microaerobically free-living cells and in nodules. Individual transposon Tn3 HoHo1 insertions in this DNA region caused a reduced nodulation competitiveness. This new symbiotic region, occupying approximately 5 kilobases of pRmeGR4b DNA, was called nfe (nodule formation efficiency). PMID- 2546915 TI - Nucleotide sequence and expression of the gene for the site-specific integration protein from bacteriophage HP1 of Haemophilus influenzae. AB - The nucleotide sequence of the leftmost 2,363 base pairs of the HP1 genome, which includes the attachment site (attP) and the integration region, was determined. This sequence contained an open reading frame encoding a 337-residue polypeptide, which is a member of the integrase family of site-specific recombination proteins as judged by sequence comparison. The open reading frame was located immediately adjacent to the att site and was oriented so that initiation of translation would begin distal to the att site and end in its immediate vicinity. Expression of this DNA segment in Escherichia coli provided extracts which promoted site specific recombination between plasmids containing cloned HP1 attP and Haemophilus influenzae attB sites. This recombination was directional, since no reaction was observed between plasmids containing attR and attL sites. The reaction was stimulated by the accessory protein integration host factor of E. coli. Evidence was also obtained that the integration host factor influenced the levels of HP1 integrase expression. The deduced amino acid sequence of HP1 integrase has remarkable similarity to that deduced for the integrase of coliphage 186. PMID- 2546916 TI - A novel sigma factor is involved in expression of the rpoH gene of Escherichia coli. AB - The Escherichia coli rpoH gene encoding sigma 32, which is involved in the heat shock response, is transcribed from as many as four promoters. We have isolated a novel sigma factor of about 24 kilodaltons that allows core RNA polymerase to transcribe preferentially from one of these promoters, rpoH3p. This promoter is known to be regulated by DnaA protein. The sigma 24 factor was isolated from a preparation of RNA polymerase by electroelution from sodium dodecyl sulfate polyacrylamide gels followed by renaturation. Expression of heat shock proteins is induced by treatments which include those that induce the stringent response. Under such conditions, decreased transcription from rpoH3p and no increase in transcription from other rpoH promoters were observed. This result suggests that induction of heat shock proteins by the stringent response is not mediated by increased transcription of the rpoH gene. PMID- 2546918 TI - Overproduction and identification of the ftsQ gene product, an essential cell division protein in Escherichia coli K-12. AB - ftsQ is an essential cell division gene in Escherichia coli. The ftsQ gene has been sequenced, and a presumptive open reading frame has been identified; however, no protein product has been observed (A.C. Robinson, D.J. Kenan, G.F. Hatfull, N.F. Sullivan, R. Spiegelberg, and W.D. Donachie, J. Bacteriol. 160:546 555, 1984, and Q.M. Yi, S. Rockenbach, J.E. Ward, Jr., and J. Lutkenhaus, J. Mol. Biol. 184:399-412, 1985). The ftsQ gene was isolated on a 970-base-pair EcoRI PvuII fragment of the E. coli chromosome and used to construct a trp-lac (Ptac) transcriptional fusion in plasmid pKK223-3. The fused construct (pDSC78) complemented an ftsQ1(Ts) mutant strain in trans, restoring growth at 42 degrees C on low-salt medium. An ftsQ1(Ts) mutant strain transformed with pDSC78 appeared normal upon microscopic examination, with no indication of filamentation. The ftsQ gene product was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and two-dimensional gel electrophoresis of radiolabeled, isopropyl-beta-D-thiogalactopyranoside-induced maxicell and normal cell extracts. As predicted from the nucleotide sequence, the 970-base-pair EcoRI-PvuII fragment encoded a polypeptide of approximately 31,400 daltons. Analysis of the data obtained from pulse-chase experiments in maxicells and normal cells suggests that the FtsQ protein is stable. Most of the radiolabeled FtsQ protein from maxicells was found in the inner membrane. On the basis of available information, the prior inability to detect FtsQ can be attributed to low levels of transcription or translation rather than to proteolysis. PMID- 2546917 TI - Expression of Escherichia coli dnaA and mioC genes as a function of growth rate. AB - The synthesis of specific cellular components related to the initiation process of DNA replication was correlated with changes in growth rate. The concentrations of DnaA protein and mioC mRNA were determined for cells grown at six different growth rates; both increased relative to either total protein or total RNA, respectively, as the growth rate increased. Expression from the chromosomal mioC promoter, which contains a DnaA protein-binding site, was not repressed when the DnaA protein concentration was increased and was not derepressed in a dnaA46 mutant at 42 degrees C. The mioC transcript had a characteristic mRNA-type half life of 1.51 min. PMID- 2546919 TI - Expression and secretion of the S-1 subunit and C180 peptide of pertussis toxin in Escherichia coli. AB - The structural gene of the S-1 subunit of pertussis toxin (rS-1) and the catalytic C180 peptide of the S-1 subunit (C180 peptide) were independently subcloned downstream of the tac promoter in Escherichia coli. Both constructions included DNA encoding for the predicted leader sequence of the S-1 subunit which was inserted between the tac promoter and the structural gene. E. coli containing the plasmids encoding for rS-1 and C180 peptide produced a peptide that reacted with anti-pertussis toxin antibody and had a molecular weight corresponding to that of the cloned gene; some degradation of rS-1 was observed. Extracts of E. coli containing plasmids encoding for rS-1 and the C180 peptide possessed ADP ribosyltransferase activity. Subcellular fractionation showed that both rS-1 and the C180 peptide were present in the periplasm, indicating that E. coli recognized the pertussis toxin peptide leader sequence. The protein sequence of the amino terminus of the C180 peptide was identical to that of authentic S-1 subunit produced by Bordetella pertussis, which showed that E. coli leader peptidase correctly processed the pertussis toxin peptide leader sequence. Two single amino acid substitutions at residue 26 (C180I-26) and residue 139 (C180S 139) which were previously shown to reduce ADP-ribosyltransferase activity were introduced into the C180 peptide. C180I-26 possessed approximately 1% of the NAD glycohydrolase activity of the C180 peptide, suggesting that tryptophan 26 functions in the interaction of NAD with the C180 peptide. In contrast, C180S-139 possessed essentially the same level of NAD-glycohydrolase activity as the C180 peptide, suggesting that glutamic acid 139 does not function in the interaction of NAD but plays a role in a later step in the ADP-ribosyltransferase reaction. PMID- 2546920 TI - Construction of TnphoA gene fusions in Rhodobacter sphaeroides: isolation and characterization of a respiratory mutant unable to utilize dimethyl sulfoxide as a terminal electron acceptor during anaerobic growth in the dark on glucose. AB - We have constructed a suicide vector, pU1800, containing the transposable element TnphoA (Tn5 IS50L::phoA), for the purpose of producing protein fusions in vivo between the Escherichia coli alkaline phosphatase (APase) and proteins of the facultative photoheterotroph, Rhodobacter sphaeroides. We introduced TnphoA into the genome of R. sphaeroides at a coupled conjugation-transposition frequency of approximately 1 x 10(-6). Fusions giving rise to APase expression, as judged by blue-colony pigmentation when exconjugants were plated on growth medium containing the chromogenic indicator 5-bromo-4-chloro-3-indolyl phosphate, were observed in about 1% of the exconjugants. Numerous, distinguishable mutant phenotypes have been generated by this method, including those which lack the ability to use dimethyl sulfoxide as a terminal electron acceptor during anaerobic respiration, as well as those which are photosynthetically incompetent or altered in pigment synthesis, and others that express resistance to chlorate. The growth and spectral characteristics of several of these mutants, as well as the localization and quantitation of subcellular APase activity under different physiological conditions, have been examined. The presence of TnphoA in the host genome has been confirmed for each mutant analyzed, and specifically tagged DNA fragments containing TnphoA have been identified and localized; cosmids containing R. sphaeroides genomic DNA capable of complementing individual mutants have also been isolated. The usefulness of this approach in studying gene activity in R. sphaeroides is discussed. PMID- 2546921 TI - Genetic characterization of the pnuC gene, which encodes a component of the nicotinamide mononucleotide transport system in Salmonella typhimurium. AB - The pnuC gene, which encodes a component of the nicotinamide mononucleotide transport system, has been mapped and oriented. The gene order of the pnuC region, which is at min 17 of the Salmonella chromosome, is nadA-pnuC-aroG-gal. Polarity tests, with pnuC::Mu d-lac operon fusions, reveal that the pnuC gene is the promoter distal gene in an operon with the nadA gene, which encodes the second enzyme of the pyridine biosynthetic pathway. The nadA pnuC operon is regulated by the NadI repressor. The pnuC gene also has its own promoter, since strains with a nadA::Tn10d(Tc) insertion still express the pnuC gene at a low, unregulated level. PMID- 2546923 TI - Intracellular PPi concentration is not directly dependent on amount of inorganic pyrophosphatase in Escherichia coli K-12 cells. AB - No correlation was observed between the level of inorganic pyrophosphatase (PPase) and the intracellular concentration of PPi in Escherichia coli cells. In exponentially growing cells the intracellular PPi concentration was in every case 1.5 nmol/mg (dry weight) or about 0.5 mM, even though the amount of PPase was varied from 15 to 2,600% of the control amount by mutation or by using a multicopy plasmid with an inserted gene (ppa) encoding PPase. The PPi concentration could, however, be increased or decreased from the control level under some stressful conditions. PMID- 2546922 TI - A mutation in the amino terminus of a hybrid TrpC-TonB protein relieves overproduction lethality and results in cytoplasmic accumulation. AB - We have developed a selection for mutations in a trpC-tonB gene fusion that takes advantage of the properties of the plasmid-encoded TrpC-TonB hybrid protein. The TrpC-TonB hybrid protein consists of amino acids 1 through 25 of the normally cytoplasmic protein, TrpC, fused to amino acids 12 through 239 of TonB. It is expressed from the trp promoter and is regulated by the trpR gene and the presence or absence of tryptophan. Under repressing conditions in the presence of tryptophan, the trpC-tonB gene can restore phi 80 sensitivity to a tonB deletion mutant, which indicates that TrpC-TonB can be exported and is functional. High level expression of TrpC-TonB protein in the absence of tryptophan results in virtually immediate cessation of growth for strains carrying the trpC-tonB plasmid. By selecting for survivors of the induced growth inhibition (overproduction lethality), we have isolated a variety of mutations. Many of the mutations decrease expression of the TrpC-TonB protein, as expected. In addition, three independently isolated mutants expressing normal levels of TrpC-TonB protein result in a Gly----Asp substitution within the hydrophobic amino terminus of TonB. The mutant proteins are designated TrpC-TonBG26D. The mutations are suppressed by prlA alleles, known to suppress export (signal sequence) mutations. TrpC-TonB proteins carrying the Gly----Asp substitution accumulate in the cytoplasm. We conclude that the Gly----Asp substitution is an export mutation. TrpC-TonBG26D protein has been purified and used to raise polyclonal antibodies that specifically recognize both TrpC-TonB protein and wild-type TonB protein. PMID- 2546924 TI - Nucleotide sequences of fic and fic-1 genes involved in cell filamentation induced by cyclic AMP in Escherichia coli. AB - The nucleotide sequences of fic-1 involved in the cell filamentation induced by cyclic AMP in Escherichia coli and its normal counterpart fic were analyzed. The open reading frame of both fic-1 and fic coded for 200 amino acids. The Gly at position 55 in the Fic protein was changed to Arg in the Fic-1 protein. The promoter activity of fic was confirmed by fusing fic and lacZ. The gene downstream from fic was found to be pabA (p-aminobenzoate). There is an open reading frame (ORF190) coding for 190 amino acids upstream from the fic gene. Computer-assisted analysis showed that Fic has sequence similarity with part of CDC28 of Saccharomyces cerevisiae, CDC2 of Schizosaccharomyces pombe, and FtsA of E. coli. In addition, ORF190 has sequence similarity with the cyclosporin A binding protein cyclophilin. PMID- 2546926 TI - Purification and characterization of membrane-bound 5'-nucleotidase of Vibrio parahaemolyticus. AB - Membrane-bound 5'-nucleotidase from Vibrio parahaemolyticus was solubilized and purified using a nonionic detergent, heptyl-beta-D-thioglucoside, and was characterized. This enzyme required Mg2+ for activity, maximum activity being observed at 5 and 20 mM Mg2+ with AMP and ATP, respectively, as substrates. Of the divalent cations tested, Mn2+ and Co2+ were able to replace Mg2+ partially, whereas Ca2+ was ineffective. Zinc strongly inhibited the enzyme activity and Ni2+ caused partial inhibition. This enzyme required Cl- for activity, the optimal concentration being 20 mM or more. The order of effectiveness of anions was Cl- greater than Br- greater than I- approximately NO3-. Sulfate and acetate were ineffective. The optimal pH was 8.0. The activity of the purified enzyme was stimulated by the addition of lipid to the assay mixture. This enzyme hydrolyzed all 5'-nucleotides tested, but did not hydrolyze 3'-nucleotides or ribose 5 phosphate. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the enzyme appeared to be a single polypeptide, with a molecular weight of 72 kDa. PMID- 2546925 TI - Rat liver phosphoribosyl pyrophosphate synthetase: existence of the purified enzyme as heterogeneous aggregates and identification of the catalytic subunit. AB - Mammalian phosphoribosyl pyrophosphate (PRPP) synthetase has been extensively investigated. However, considerable ambiguity remains concerning its physical and regulatory properties. We purified PRPP synthetase from rat liver and studied some of the physical properties, in parallel with cloning experiments (Taira, M. et. al. [1987] J. Biol. Chem. 262, 14867-14870). 1) The enzyme was purified to a specific activity of 7,280 milliunits/mg, the highest value in the literature for a mammalian PRPP synthetase. The apparent molecular mass was over 1,000 kDa. 2) The final preparation contained 34-kDa, 38-kDa, and 40-kDa protein species, as analyzed by SDS gel electrophoresis. 3) Further attempts at separation using conventional procedures only led to a co-purification of the components. Thus, the purified enzyme appears to exist as complex aggregates composed of heterogeneous components. 4) Gel filtration of the enzyme in the presence of 1 M MgCl2 isolated part of the 34-kDa component, free of other species. The preparation was catalytically active, indicating that this component is the catalytic subunit. 5) The amino acid composition of the 34-kDa subunit and the amino acid sequences of its N-terminal region and of two tryptic peptides were determined. The results are in accord with the results of cDNA analyses. PMID- 2546927 TI - Effect of internally loaded iodide, thiocyanate, and perchlorate on sodium dependent iodide uptake by phospholipid vesicles reconstituted with thyroid plasma membranes: iodide counterflow mediated by the iodide transport carrier. AB - Na+-dependent I- transport and I- counterflow were studied using phospholipid vesicles (P-vesicles) made of porcine thyroid plasma membranes and soybean phospholipid by sonication. 1) I- uptake by P-vesicles incubated in the presence of external Na+ was higher than that by P-vesicles incubated in choline+ instead of Na+. The vesicles exhibited Na+-dependent I- uptake. When P-vesicles were internally loaded with I- prior to incubation in Na+, a further increase in Na+ dependent I- uptake was observed, although the concentration of internal I- was very much higher than that outside. In the absence of external Na+, I- uptake by P-vesicles preloaded with I- was comparable to baseline uptake. 2) Na+-dependent I- uptake by P-vesicles not loaded with I- and enhanced Na+-dependent I- uptake by P-vesicles preloaded with I- were both inhibited by either of SCN- and ClO4- added outside the vesicles. 3) When P-vesicles were loaded with SCN- instead of I and incubated in Na+, I- uptake by these vesicles was also higher than baseline Na+-dependent I- uptake. However, a ClO4- load did not result in an increase in I uptake. These results indicate that Na+-dependent I- transport including Na+ dependent I- counterflow is specifically mediated by the thyroid I- carrier. SCN- - I- counterflow in addition to I- - I- counterflow occurs dependently on Na+, but ClO4- - I- counterflow does not. PMID- 2546928 TI - Affinity separation of human plasma gelsolin on Affi-Gel Blue. AB - Human plasma gelsolin was specifically eluted with 1 mM adenosine 5'-triphosphate from an Affi-Gel Blue column. Since the ionic strength of sodium chloride required to elute the protein from the dye column was much higher than that of 1 mM adenosine 5'-triphosphate, the binding of plasma gelsolin with the dye-ligand appeared to be biospecific. Taking advantage of this affinity interaction, we have developed a revised purification method of human plasma gelsolin. The purification included ammonium sulfate precipitation, diethylaminoethyl-Sepharose chromatography, Affi-Gel Blue chromatography, and Phenyl-Sepharose chromatography. The method allowed a reproducible purification of the protein to apparent homogeneity, producing a 331-fold purification with a yield of 6%. PMID- 2546929 TI - Cloning and expression of the 5'-nucleotidase gene of Vibrio parahaemolyticus in Escherichia coli and overproduction of the enzyme. AB - The gene encoding the membrane-bound 5'-nucleotidase of Vibrio parahaemolyticus was cloned and expressed in Escherichia coli. Cells of E. coli harboring a plasmid, pNUT5, which carries the 5'-nucleotidase gene were able to grow on ATP as the sole source of carbon, although the original cells were not. The 5' nucleotidase activity was detected in whole cells of E. coli harboring pNUT5 and in membrane vesicles prepared from these cells. Most properties of the 5' nucleotidase produced in E. coli, that is, its requirements for Cl- and Mg2+, substrate specificity, and inhibition by Zn2+, were similar to those observed in V. parahaemolyticus, but some alterations in properties were observed: The 5' nucleotidase was partially inducible in V. parahaemolyticus, but its expression in E. coli was completely constitutive. The specific activity of the 5' nucleotidase in membrane vesicles of E. coli harboring the plasmid was 30 times that observed in whole cells, whereas the specific activities in membrane vesicles and in whole cells of V. parahaemolyticus were almost the same. A new, dense band of protein with an apparent molecular mass of 63 kDa was detected when membrane proteins of E. coli harboring the plasmid were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PMID- 2546930 TI - The unraveling architecture of the junctional sarcoplasmic reticulum. AB - The sarcoplasmic reticulum (SR) of skeletal muscle controls the contraction relaxation cycle by raising and lowering the myoplasmic free-Ca2+ concentration. The coupling between excitation, i.e., depolarization of sarcolemma and transverse tubule (TT) and Ca2+ release from the terminal cisternae (TC) of SR takes place at the triad. The triad junction is formed by a specialized region of the TC, the junctional SR, and the TT. The molecular architecture and protein composition of the junctional SR are under active investigation. Since the junctional SR plays a central role in excitation-contraction coupling and Ca2+ release, some of its protein constituents are directly involved in these processes. The biochemical evidence supporting this contention is reviewed in this article. PMID- 2546931 TI - The muscle ryanodine receptor and its intrinsic Ca2+ channel activity. AB - In skeletal and cardiac muscle, contraction is initiated by the rapid release of Ca2+ ions from the intracellular membrane system, sarcoplasmic reticulum. Rapid mixing vesicle ion flux and planar lipid bilayer-single-channel measurements have shown that Ca2+ release is mediated by a high-conductance, ligand-gated Ca2+ channel. Using the Ca2+ release-specific probe ryanodine, a 30 S protein complex composed of four polypeptides of Mr approximately 400,000 has been isolated. Reconstitution of the purified skeletal and cardiac muscle 30 S complexes into planar lipid bilayers induced single Ca2+ channel currents with conductance and gating kinetics similar to those of native Ca2+ release channels. Electron microscopy revealed structural similarity with the protein bridges ("feet") that span the transverse-tubule-sarcoplasmic reticulum junction. These results suggest that striated muscle contains an intracellular Ca2+ release channel that is identical with the ryanodine receptor and the transverse-tubule-sarcoplasmic reticulum spanning feet structures. PMID- 2546932 TI - Inositol trisphosphate and excitation-contraction coupling in skeletal muscle. AB - The role of inositol trisphosphate as a chemical messenger in excitation contraction coupling is discussed, both in terms of positive and negative results. The evidence presented includes experiments on the effect of inositol trisphosphate in intact and skinned fibers, in calcium release from isolated sarcoplasmic reticulum vesicles, in activation of single calcium release channels incorporated in planar bilayers, and biochemical experiments that have established the presence of all the intermediate steps involved in the metabolism of phosphoinositides, both in intact muscle and in isolated membranes. From these results, it is clear that a role for inositol triphosphate in skeletal muscle function is highly likely; whether this molecule is the physiological messenger in excitation-contraction coupling remains to be established. PMID- 2546934 TI - Common sites for recombination and cleavage mediated by bacteriophage T4 DNA topoisomerase in vitro. AB - We have previously shown that purified T4 DNA topoisomerase promotes illegitimate recombination between two lambda DNA molecules, or between lambda and plasmid DNA in vitro (Ikeda, H. (1986) Proc. Natl. Acad. Sci. U. S. A. 83, 922-926). Since the recombinant DNA contains a duplication or deletion, it is inferred that the cross-overs take place between nonhomologous sequences of lambda DNA. In this paper, we have examined the sequences of the recombination junctions produced by the recombination between two lambda DNA molecules mediated by T4 DNA topoisomerase. We have shown that there is either no homology or there are 1-5 base pair homologies between the parental DNAs in seven combinations of lambda recombination sites, indicating that homology is not essential for the recombination. Next, we have shown an association of the recombination sites with the topoisomerase cleavage sites, indicating that a capacity of the topoisomerase to make a transient double-stranded break in DNA plays a role in the illegitimate recombination. A consensus sequence for T4 topoisomerase cleavage sites, RNAY decreases NNNNRTNY, was deduced. The cleavage experiment showed that T4 topoisomerase-mediated cleavage takes place in a 4-base pair staggered fashion and produces 5'-protruding ends. PMID- 2546935 TI - Dephosphorylation of neuromodulin by calcineurin. AB - Neuromodulin (p57, GAP-43, F1, B-50) is a major neural-specific, calmodulin binding protein found in brain, spinal cord, and retina that is associated with membranes. Phosphorylation of neuromodulin by protein kinase C causes a significant reduction in its affinity for calmodulin (Alexander, K. A., Cimler, B. M., Meirer, K. E., and Storm, D. R. (1987) J. Biol. Chem. 262, 6108-6113). It has been proposed that neuromodulin may function to bind and concentrate calmodulin at specific sites within neurons and that activation of protein kinase C causes the release of free calmodulin at high concentrations near its target proteins. It was the goal of this study to determine whether bovine brain contains a phosphoprotein phosphatase that will utilize phosphoneuromodulin as a substrate. Phosphatase activity for phosphoneuromodulin was partially purified from a bovine brain extract using DEAE-Sephacel and Sephacryl S-200 gel filtration chromatography. The neuromodulin phosphatase activity was resolved into two peaks by Affi-Gel Blue chromatography. One of these phosphatases, which represented approximately 60% of the total neuromodulin phosphatase activity, was tentatively identified as calcineurin by its requirement for Ca2+ and calmodulin (CaM) and inhibition of its activity by chlorpromazine. Therefore, bovine brain calcineurin was purified to homogeneity and examined for its phosphatase activity against bovine phosphoneuromodulin. Calcineurin rapidly dephosphorylated phosphoneuromodulin in the presence of micromolar Ca2+ and 3 microM CaM. The apparent Km and Vmax for the dephosphorylation of neuromodulin, measured in the presence of micromolar Ca2+ and 2 microM CaM, were 2.5 microM and 70 nmol Pi/mg/min, respectively, compared to a Km and Vmax of 4 microM and 55 nmol Pi/mg/min, respectively, for myosin light chain under the same conditions. Dephosphorylation of neuromodulin by calcineurin was stimulated 50-fold by calmodulin in the presence of micromolar free Ca2+. Half-maximal stimulation was observed at a calmodulin concentration of 0.5 microM. We propose that phosphoneuromodulin may be a physiologically important substrate for calcineurin and that calcineurin and protein kinase C may regulate the levels of free calmodulin available in neurons. PMID- 2546936 TI - Ca2+-dependent and cAMP-dependent control of nicotinic acetylcholine receptor phosphorylation in muscle cells. AB - Mouse BC3H1 myocytes were incubated with 32Pi before acetylcholine receptors were solubilized, immunoprecipitated, and subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis. More than 90% of the 32P found in the receptor was bound to the delta subunit. Two phosphorylation sites in this subunit were resolved by reverse phase high performance liquid chromatography after exhaustive proteolysis of the protein with trypsin. Sites 1 and 2 were phosphorylated to approximately the same level in control cells. The divalent cation ionophore, A23187, increased 32P in site 1 by 40%, but did not affect the 32P content of site 2. In contrast, isoproterenol increased 32P in site 2 by more than 60%, while increasing 32P in site 1 by only 20%. When dephosphorylated receptor was incubated with [gamma-32P]ATP and the catalytic subunit of cAMP dependent protein kinase, the delta subunit was phosphorylated to a maximal level of 1.6 phosphates/subunit. Approximately half of the phosphate went into site 2, with the remainder going into a site not phosphorylated in cells. The alpha subunit was phosphorylated more slowly, but phosphorylation of both alpha and delta subunits was blocked by the heat-stable protein inhibitor of cAMP-dependent protein kinase. Phosphorylation of the receptor was also observed with preparations of phosphorylase kinase. In this case phosphorylation occurred in the beta subunit and site 1 of the delta subunit, neither of which were phosphorylated by cAMP-dependent protein kinase. The rate of receptor phosphorylation by phosphorylase kinase was slow relative to that catalyzed by cAMP-dependent protein kinase. Therefore, it can not yet be concluded that phosphorylase kinase phosphorylates the beta subunit and the delta subunit site 1 in cells. However, the results strongly support the hypothesis that phosphorylation by cAMP-dependent protein kinase accounts for phosphorylation of the alpha subunit and the delta subunit site 2 in response to elevations in cAMP. PMID- 2546937 TI - Characterization of the bradykinin-stimulated calcium influx pathway of cultured vascular endothelial cells. Saturability, selectivity, and kinetics. AB - Measurement of fura-2 fluorescence and 45Ca2+ uptake was used to evaluate Ca2+ influx in cultured bovine aortic endothelial cells (BAECs) stimulated by bradykinin (BK). The BK-stimulated influx pathway was characterized with respect to its 1) sensitivity to extracellular Ca2+, 2) inhibition by membrane depolarization, and 3) permeability to Ba2+ and Sr2+. The results indicate that the activity of the influx pathway is a saturable function of extracellular Ca2+ and that membrane depolarization inhibits Ca2+ influx by changing the apparent affinity and maximal capacity of the pathway for Ca2+. Fura-2 fluorescence was used to compare the profiles for BK-stimulated changes in cytosolic Ca2+, Sr2+, and Ba2+ (Ca2+i, Ba2+i, and Sr2+i). Addition of Ca2+ and Sr2+ to Ca2+-depleted cells in the presence of BK produced a transient increase in Ca2+i and Sr2+i. Following the peak of the response, Ca2+i and Sr2+i declined within 2 min to a steady elevated level. Blockade of influx by the addition of La3+ at the peak of the response to Ca2+ and Sr2+ immediately reduced Ca2+i and Sr2+i to basal levels. Addition of Ba2+ to Ca2+-depleted cells in the presence of BK produced an increase in Ba2+i which continued to rise with time to a steady level. Addition of La3+ after Ba2+, however, did not reduce Ba2+i. These results suggest that 1) Ca2+ and Sr2+ (but not Ba2+) are sequestered by intracellular mechanisms and that the declining phase of the Ca2+ and Sr2+ response reflects a time and divalent cation-dependent inactivation of the influx pathway. The inactivation of the influx pathway was further demonstrated by measuring the kinetics of BK stimulated 45Ca2+ uptake into BAECs. The results of these experiments demonstrate that BK stimulates a 100- to 150-fold increase in Ca2+ permeability of the BAEC but that the influx pathway turns off or inactivates within 2 min. The magnitude of the flux, the voltage sensitivity, and the ability to conduct Ca2+, Sr2+, and Ba2+ are suggestive of a channel mechanism. PMID- 2546938 TI - The roles of cAMP and cAMP-dependent protein kinase in the expression of cholesterol side chain cleavage and steroid 11 beta-hydroxylase genes in mouse adrenocortical tumor cells. AB - The expression of the genes encoding cholesterol side chain cleavage enzyme (SCC) and steroid 11 beta-hydroxylase (11 beta-OHase) was examined in Y1 mouse adrenocortical tumor cells and in derivative cell lines defective in cAMP dependent protein kinase activity. Y1 cells expressed both genes constitutively, and treatment with 8-bromo-cAMP (8-Br-cAMP) increased expression 5-10-fold. In three independent protein kinase mutants, expression of SCC and 11 beta-OHase was impaired to degrees dependent upon the severity of defect in cAMP-dependent protein kinase activity. In Kin-2, the least impaired mutant clone, basal expression of SCC was the same as in Y1 cells. Treatment of Kin-2 with 8-Br-cAMP increased SCC RNA to the levels seen in stimulated Y1 cells. In contrast, clone Kin-8, the most severe mutant, expressed markedly diminished basal and 8-Br-cAMP stimulated levels of SCC mRNA. Kin-7 had basal and 8-Br-cAMP-stimulated levels of SCC mRNA which were intermediate to Kin-2 and Kin-8. None of the Kin mutants constitutively expressed detectable levels of 11 beta-OHase transcripts, and only Kin-2 responded to treatment with 8-Br-cAMP with increased expression of 11 beta OHase; however, the time course of induction in Kin-2 was significantly delayed. The disparate patterns of expression of SCC and 11 beta-OHase in the Kin mutants suggest that these genes differ in their absolute requirement for cAMP-dependent protein kinase activity. Experiments also were performed in which Kin-7 and Kin-8 mutants were restored to cAMP-responsive states by transfection with genes encoding normal sub-units of cAMP-dependent protein kinase. These phenotypic revertants recovered 8-Br-cAMP-inducible expression of SCC and 11 beta-OHase. These results strongly support the hypothesis that impaired expression of steroidogenic enzymes in the Kin mutants results directly from defects in cAMP dependent protein kinase activity. PMID- 2546933 TI - Pharmacology of calcium release from sarcoplasmic reticulum. AB - Calcium release from sarcoplasmic reticulum (SR) has been elicited in response to additions of many different agents. Activators of Ca2+ release are here tentatively classified as activators of a Ca2+-induced Ca2+ release channel preferentially localized in SR terminal or as likely activators of other Ca2+ efflux pathways. Some of these pathways may be associated with several different mechanisms for SR Ca2+ release that have been postulated previously. Studies of various inhibitors of excitation-contraction coupling and of certain forms of SR Ca2+ release are summarized. The sensitivity of isolated SR to certain agents is unusually affected by experimental conditions. These effects can seriously undermine attempts to anticipate effects of the same pharmacological agents in situ. Finally, mention is made of a new preparation ("sarcoballs") designed to make the pharmacological study of SR Ca2+ release more accessible to electrophysiologists, and some concluding speculations on the future of SR pharmacology are offered. PMID- 2546939 TI - Thyroglobulin-mediated one- and two-electron oxidations of glutathione and ascorbate in thyroid peroxidase systems. AB - The one- or two-electron oxidation of thyroglobulin by the thyroid peroxidase system was found to be regulated by the iodine content of thyroglobulin. The catalytic intermediate of thyroid peroxidase observed at steady state of the reaction was Compound I and II when the iodine content in thyroglobulin was 0.2 and 0.7%, respectively, apparent rate constants for the rate-limiting steps being estimated at 4.7 x 10(7) and 4.8 x 10(4) M-1 S-1. The thyroglobulin-mediated oxidation of GSH occurred by way of two-electron transfer at 0.2% iodine content and by way of one-electron transfer at 0.7% iodine content. The spin-trapping experiment with 5,5-dimethyl-1-pyrroline-N-oxide showed that glutathione radicals were formed in the latter reaction but not in the former. In the reactions of thyroid peroxidase, the one- and two-electron oxidations of ascorbate were also mediated by 0.2 and 0.7% iodine thyroglobulins, respectively. The reactions were analyzed and mimicked with the use of p-cresol and p-acetaminophenol as a mediator in the reactions of lactoperoxidase and thyroid peroxidase. PMID- 2546940 TI - A novel fetal insulin-like growth factor (IGF) I receptor. Mechanism for increased IGF I- and insulin-stimulated tyrosine kinase activity in fetal muscle. AB - Insulin and insulin-like growth factor (IGF) I receptors from fetal and adult rat skeletal muscle were compared in order to gain insight into the evolving functions of the hormones during development. Basal, insulin-stimulated, and IGF I-stimulated receptor phosphorylation and tyrosine kinase activity are severalfold higher in partially purified receptor preparations from fetal muscle in comparison with equal numbers of receptors from adult muscle. There are distinct insulin and IGF I receptors with Mr 95,000 beta subunits in adult muscle, as evidenced by hormone dose-response curves, immunoprecipitation with specific antibodies, binding to insulin and IGF I affinity columns, and analysis of tryptic phosphopeptides. In addition to these two receptor species, fetal muscle contains a receptor with a Mr 105,000 beta subunit. The fetal receptor is structurally more closely related to the IGF-I receptor than the insulin receptor on the basis of its precipitation with specific antibodies, binding to an IGF I affinity column, and tryptic phosphopeptide map. The fetal receptor does not appear to bind insulin but, unlike the IGF-I receptor, its phosphorylation is stimulated by low physiological concentrations of both insulin and IGF I. This could be explained by the cross-phosphorylation of fetal receptors by activated insulin receptors. Expression of the fetal receptor is highest in the fetus and decreases markedly during the first 2 weeks of postnatal life. The fetal receptor appears to account for the high tyrosine kinase activity of fetal muscle and may be an important mediator of responses to both insulin and IGF I early in development. PMID- 2546941 TI - Internalization and activation of the rat liver insulin receptor kinase in vivo. AB - The preparation of clearly delineated plasmalemma (PM) and endosomal subcellular fractions from rat liver has allowed us to compare insulin receptor (IR) kinase activity at the cell surface and in hepatic endosomes (ENs) as a function of dose and time after injected insulin. Tyrosine kinase activity in PM and ENs was measured, after solubilization and partial purification by wheat germ agglutinin chromatography (lectin-purified), using poly(Glu:Tyr) as substrate. Following the injection of a subsaturating dose of insulin (1.5 micrograms/100 g body weight), lectin-purified receptor showed peak activation at 30 s in PM and at 2 min in ENs. As observed previously (Khan, M. N., Savoie, S., Bergeron, J. J. M., and Posner, B. I. (1986) J. Biol. Chem. 261, 8462-8472) autophosphorylation activity was also augmented following insulin injection. In a pattern virtually identical to that of exogenous kinase activity, autophosphorylation attained peak activity at 30 s in PM and at 2 min in ENs. The time course of IR autophosphorylation in intact membranes was very similar to that observed for lectin purified receptors and was seen with an injected insulin dose as low as 150 ng/100 g body weight. Phosphatase treatment of the solubilized endosomal receptor abolished its enhanced activity. Hence, insulin treatment led to in vivo receptor phosphorylation which was reflected in the enhancement of both tyrosine kinase and autophosphorylation activities. Significant differences in the phosphorylation activities of PM and ENs were observed. Phosphoamino acid analyses revealed that the activated IR of intact PM was autophosphorylated in vitro, at both serine (55%) and tyrosine (45%) residues; whereas the activated IR of intact ENs was phosphorylated in vitro exclusively on tyrosine autophosphorylation specific activity for the activated IR of ENs was 3- to 4 fold that of the IR of PM. This was observed for the lectin purified IRs as well as for IRs of intact cell fractions. The reduced level of IR autophosphorylation in PM was not due to occlusion of tyrosine acceptor sites by prior in vivo phosphorylation. The rapidity with which activated IR accumulates in ENs as well as the sensitivity of endosomal IR kinase to activation by injected insulin are consistent with the endosomal apparatus serving a physiologically significant site for the regulation of transmembrane signaling. PMID- 2546942 TI - The active site of the thrombin-thrombomodulin complex. A fluorescence energy transfer measurement of its distance above the membrane surface. AB - The location of the active site of the membrane-bound anticoagulant complex of thrombin and thrombomodulin has been determined relative to the membrane surface using fluorescence energy transfer. Thrombin was reacted with 5-(dimethylamino)-1 naphthalenesulfonylglutamylglycylarginyl chloromethyl ketone (DEGR-CK) to yield DEGR-thrombin, an analogue of thrombin with a fluorescent dye covalently attached to its active site. When DEGR-thrombin was titrated with thrombomodulin that had been reconstituted into phospholipid vesicles containing octadecylrhodamine, singlet-singlet energy transfer was observed between the donor dyes, each in an active site of a DEGR-thrombin bound to thrombomodulin, and the acceptor dyes at the outer surface of the phospholipid bilayer. The extent of energy transfer reached a maximum when DEGR-thrombin and thrombomodulin were equimolar in the sample, as expected for the formation of a 1:1 complex between thrombin and thrombomodulin. This energy transfer was dependent upon the binding of DEGR thrombin to thrombomodulin because no energy transfer was observed with vesicles that lacked thrombomodulin, and the extent of energy transfer was reduced greatly by the addition of excess unmodified nonfluorescent thrombin to compete with DEGR thrombin for binding to the thrombomodulin. From the dependence of the energy transfer upon the acceptor density and assuming kappa 2 = 2/3, the distance of closest approach between a dye in the active site of the thrombin-thrombomodulin complex and a dye at the membrane surface was determined to average 66 A (65 +/- 3 A for phosphatidylcholine vesicles without and 67 +/- 5 A for those with 20% phosphatidylserine). This distance was also insensitive to the presence or absence of Ca2+. These direct measurements indicate that the active site of the membrane-bound thrombin-thrombomodulin complex is located far above the phospholipid surface, that the peptide bond cleaved during the activation of protein C is situated about 66 A above the membrane, that the thrombin binding site on thrombomodulin is positioned more than 45 A above the membrane, ant that thrombin, with a diameter near 40 A, is not positioned alongside thrombomodulin near the membrane to form the thrombin-thrombomodulin complex but is instead bound "on top" of thrombomodulin. PMID- 2546943 TI - Iron ion-dependent modification of bases in DNA by the superoxide radical generating system hypoxanthine/xanthine oxidase. AB - Damage to the bases in DNA produced by the hypoxanthine/xanthine oxidase system in the presence of iron ions was studied. The base products in DNA were measured using gas chromatography-mass spectrometry with selected ion monitoring after acidic hydrolysis of DNA and trimethylsilylation. Products identified were cytosine glycol, thymine glycol, 5,6-dihydroxycytosine, 4,6-diamino-5 formamidopyrimidine, 8-hydroxyadenine, 2,6-diamino-4-hydroxy-5 formamidopyrimidine, and 8-hydroxyguanine. These are typical hydroxyl radical induced products of the bases in DNA. 2,6-Diamino-4-hydroxy-5-formamidopyrimidine was the major product, followed by 8-hydroxyguanine, in DNA treated with hypoxanthine/xanthine oxidase/Fe3+-EDTA. The use of Fe3+ did not cause as much damage to the bases in DNA as did the use of Fe3+-EDTA. In both systems, the formation of the products was inhibited by superoxide dismutase, catalase, dimethyl sulfoxide, mannitol, and desferrioxamine, but inhibitions were much stronger in the systems containing EDTA. Hence formation of hydroxyl radicals by a superoxide radical-assisted Fenton reaction is proposed to account for the results obtained. 2,6-Diamino-4-hydroxy-5-formamidopyrimidine, 5,6 dihydroxycytosine, 4,6-diamino-5-formamidopyrimidine, and 8-hydroxyguanine were proposed as the products in DNA to measure if one aims to measure DNA products as indices of oxidative DNA damage involving hydroxyl radicals in vivo. PMID- 2546944 TI - A permissive role of pertussis toxin substrate G-protein in P2-purinergic stimulation of phosphoinositide turnover and arachidonate release in FRTL-5 thyroid cells. Cooperative mechanism of signal transduction systems. AB - Extracellular ATP and other purinergic agonists were found to inhibit cAMP accumulation by depressing adenylate cyclase as an "inhibitory action" and/or to stimulate arachidonate release in association with phospholipase C or A2 activation and Ca2+ mobilization as "stimulatory actions" in FRTL-5 cells. The stimulatory actions of a group of P2-agonists represented by ATP were partially inhibited by the pretreatment of the cells with islet-activating protein (IAP), pertussis toxin, even when an about 41-kDa membrane protein(s) was completely ADP ribosylated. Only the IAP-sensitive part of the stimulatory actions was antagonized by 1,3-diethyl-8-phenylxanthine (DPX), an adenosine antagonist. GTP and 8-bromoadenosine 5'-triphosphate (Br-ATP) at two to three orders of higher concentrations than ATP also exerted the stimulatory actions, although they were entirely insensitive to both IAP and DPX. Ligand binding experiments with, [35S]ATP gamma S and [3H]DPX showed that ATP occupies both DPX-sensitive and insensitive receptor sites, whereas GTP does only ATP-displaceable DPX insensitive sites. Thus, lack of sensitivity of GTP action to DPX was associated with its inability to occupy the DPX-sensitive sites. Adenosine 5'-O-(1 thiotriphosphate) (ATP alpha S), adenosine 5'-O-(2-thiodiphosphate) (ADP beta S) and P1-agonists such as AMP and N6-(L-2-phenylisopropyl-adenosine (PIA) did not show any stimulatory action. Nevertheless, the agonists remarkably enhanced the stimulatory actions of GTP or Br-ATP. Such permissive actions of PIA and others were sensitive to both IAP and DPX, as were shown for a part of the stimulatory actions of ATP as well as the "inhibitory actions" of both PIA and ATP. We conclude that an IAP substrate G-protein(s) which mediates the inhibitory action of purinergic agonists via a DPX-sensitive purinergic receptor(s) may not directly link to the phospholipase C or A2 system but enhance the system which links to a DPX-insensitive P2-receptor, in an indirect or permissive manner. PMID- 2546945 TI - Characterization of the helicase and primase activities of the 63-kDa component of the bacteriophage T7 gene 4 protein. AB - Leading and lagging strand DNA synthesis at the replication fork of bacteriophage T7 DNA requires the helicase and primase activities of the gene 4 protein. Gene 4 protein consists of two colinear polypeptides of 56- and 63-kDa molecular mass. We have demonstrated previously that the 56-kDa protein possesses helicase but lacks primase activity (Bernstein, J. A., and Richardson, C. C. (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 396-400). The 63-kDa gene 4 protein has now been purified from extracts of T7-infected cells. The preparation contains 5-10% contaminating 56-kDa protein, as shown by Western analysis using polyclonal antibodies to the purified 56-kDa protein. The 63-kDa protein catalyzes DNA dependent dTTP hydrolysis and has helicase activity; both specific activities are similar to those determined for the 56-kDa protein. The 63-kDa protein efficiently synthesizes sequence-specific di-, tri-, and tetraribonucleotides and stimulates the elongation of tetraribonucleotides by T7 DNA polymerase. Although the 56-kDa protein alone lacks primase activity, it enhances the primase activity of the 63-kDa protein 4-fold. This stimulation can be accounted for by a similar increase in the amount of primers synthesized by the 63-kDa protein in the presence of the 56-kDa protein. PMID- 2546946 TI - Glucocorticoids induce the accumulation of novel angiotensinogen gene transcripts. AB - Previous work from this laboratory has shown that glucocorticoids and (Sp)-cAMPS, a cyclic AMP analogue, stimulate the accumulation of angiotensinogen mRNA in isolated hepatocytes. The present study demonstrated that glucocorticoids stimulate the accumulation of a novel, 2.25-kilobase transcript of the angiotensinogen gene, both in isolated hepatocytes and in the intact liver. (Sp) cAMPS was synergistic with dexamethasone in causing the accumulation of the larger species of RNA in hepatocytes, but had no effect by itself. Primer extension analysis and S1 mapping experiments proved that the 2.25-kilobase angiotensinogen RNA consisted of two larger forms of angiotensinogen RNA extended at their 5' ends. The novel transcripts are generated by the use of two new transcription initiation sites in the angiotensinogen gene, located at nucleotide positions -328 and -386 relative to the start site at position +1 used in the absence of hormone. A consensus TATA box is found at the expected position 25-30 nucleotides upstream from the start site at position -328, and another TATA-like sequence is found 25-30 nucleotides upstream from the start site at -386. In addition, two consensus glucocorticoid response elements occur upstream from the initiation sites. The larger angiotensinogen RNAs do not appear to code for a novel form of the angiotensinogen protein. It appears that glucocorticoids can direct transcription from a second promoter in the angiotensinogen gene and that this promoter is absolutely dependent on the hormone. To date, this situation appears to be unique to the angiotensinogen gene. PMID- 2546947 TI - The actions of Neurospora endo-exonuclease on double strand DNAs. AB - Neurospora crassa endo-exonuclease, an enzyme implicated in recombinational DNA repair, was found previously to have a distributive endonuclease activity with a high specificity for single strand DNA and a highly processive exonuclease activity. The activities of endo-exonuclease on double strand DNA substrates have been further explored. Endo-exonuclease was shown to have a low bona fide endonuclease activity with completely relaxed covalently closed circular DNA and made site-specific breaks in linear double strand DNA at a low frequency while simultaneously generating a relatively high level of single strand breaks (nicks) in the DNA. Sequencing at nicks induced by endo-exonuclease in pBR322 restriction fragments showed that the highest frequency of nicking occurred at the mid-points of two sites with the common sequence, p-AGCACT-OH. In addition, sequencing revealed less frequent nicking at identical or homologous hexanucleotide sequences in all other 54 cases examined where these sequences either straddled the break site itself or were within a few nucleotides on either side of the break site. The exonucleolytic action of endo-exonuclease on linear DNA showed about 100-fold preference for acting in the 5' to 3' direction. Removal of the 5' terminal phosphates substantially reduced this activity, internal nicking, and the ability of endo-exonuclease to generate site-specific double strand breaks. On the other hand, nicking of the dephosphorylated double strand DNA with pancreatic DNase I stimulated the exonuclease activity by almost 5-fold, but no stimulation was observed when the DNA was nicked by Micrococcal nuclease. Thus, 5'-p termini either at double strand ends or at nicks in double strand DNA are entry points to the duplex from which endo-exonuclease diffuses linearly or "tracks" in the 5' to 3' direction to initiate its major endo- and exonucleolytic actions. The results are interpreted to show how it is possible for endo exonuclease to generate single strand DNA for switching into a homologous duplex either at a nick or while remaining bound at a double strand break in the DNA. Such mechanisms are consistent with current models for recombinational double strand break repair in eukaryotes. PMID- 2546948 TI - Carotenoid biosynthesis in photosynthetic bacteria. Genetic characterization of the Rhodobacter capsulatus CrtI protein. AB - Carotenoids are photoprotective pigments present in many photosynthetic and nonphotosynthetic organisms. The desaturation of phytoene into phytofluene is an early step in the biosynthetic pathway that in the photosynthetic bacterium Rhodobacter capsulatus is mediated by the product of the crtI gene. Here we report the sequence of this gene and the identification of CrtI as a membrane protein of approximate Mr 60,000. Mutant strains with 5-fold lower or 10-fold higher levels of CrtI with respect to wild type have only small differences in their carotenoid content, indicating that the cellular concentration of CrtI is not a limiting factor in carotenoid biosynthesis. However, a correlation was found between the levels of CrtI and the formation of a photosynthetic antenna system. PMID- 2546949 TI - Insulin-like growth factor I receptor beta-subunit heterogeneity. Evidence for hybrid tetramers composed of insulin-like growth factor I and insulin receptor heterodimers. AB - In both NIH3T3 cells and HepG2 cells, insulin-like growth factor I (IGF-I) receptors possess two beta-subunits that display different electrophoretic mobilities. Increasing concentrations of IGF-I stimulated the phosphorylation of both beta-subunits to a similar extent, whereas insulin stimulated the phosphorylation of both subunits only at elevated concentrations. Both beta subunits were immunoprecipitated with p5, an insulin receptor-specific anti peptide antibody, or with A410, a polyclonal anti-insulin receptor antisera. However, if the tetrameric IGF-I receptor was first dissociated into alpha-beta heterodimers with 1 mM dithiothreitol, only the lower molecular weight beta subunit was immunoprecipitated. These results suggested that p5 and A410 specifically recognized the lower molecular weight beta-subunit but immunoprecipitated the higher molecular weight beta-subunit because it was present in the same disulfide linked tetramer. Similarly, alpha-IR-3, an antibody specific for the alpha-subunit of the IGF-I receptor, immunoprecipitated both types of beta-subunit from the intact tetramer but only the higher molecular weight beta-subunit from the dissociated heterodimers, suggesting that there are two types of alpha-subunits in the same tetramer and that the alpha-subunit recognized by alpha-IR-3 is only associated with the higher molecular weight beta subunit. Tryptic phosphopeptide maps of the lower molecular weight beta-subunit of IGF-I receptor were different from the higher molecular weight beta-subunit, but were similar to those of the insulin receptor beta-subunit. Thus, by immunochemical cross-reactivity and structural criteria, the lower molecular weight beta-subunit of the IGF-I receptor was similar to the beta-subunit of insulin receptor. These data suggest that there exists a species of IGF-I receptor that is a hybrid composed of an insulin receptor alpha-beta heterodimer and an IGF-I receptor alpha-beta heterodimer. The existence of such a hybrid receptor could have important functional consequences. PMID- 2546950 TI - Structural inferences for the native skeletal muscle sodium channel as derived from patterns of endogenous proteolysis. AB - The alpha subunit (Mr approximately 260,000) of the rat skeletal muscle sodium channel is sensitive to cleavage by endogenous proteases during the isolation of muscle surface membrane. Antisera against synthetic oligopeptides were used to map the resultant fragments in order to identify protease-sensitive regions of the channel's structure in its native membrane environment. Antibodies to the amino terminus labeled major fragments of Mr approximately 130,000 and 90,000 and lesser amounts of other peptides as small as Mr approximately 12,000. Antisera to epitopes within the carboxyl-terminal half of the primary sequence recognized two fragments of Mr approximately 110,000 and 78,000. Individual antisera also selectively labeled smaller polypeptides in the most extensively cleaved preparations. The immunoreactivity patterns of monoclonal antibodies previously raised against the purified channel were then surveyed. The binding sites for one group of monoclonals, including several that recognize subtype-specific epitopes in the channel structure, were localized within a 12-kDa fragment near the amino terminus. The distribution of carbohydrate along the primary structure of the channel was also assessed by quantitating 125I-wheat germ agglutinin and 125I concanavalin A binding to the proteolytic peptides. Most of the carbohydrate detected by these lectins was located between 22 and 90 kDa from the amino terminus of the protein. No lectin binding was detected to fragments arising from carboxyl-terminal half of the protein. These results were analyzed in terms of current models of sodium channel tertiary structure. In its normal membrane environment, the skeletal muscle sodium channel appears sensitive to cleavage by endogenous proteases in regions predicted to link the four repeat domains on the cytoplasmic side of the membrane while the repeat domains themselves are resistant to proteolysis. PMID- 2546951 TI - Cloning of murine gelsolin and its regulation during differentiation of embryonal carcinoma cells. AB - The regulation of gelsolin levels during differentiation of the murine embryonal carcinoma cell line, PC-13, was investigated using nucleic acid and immunological probes. A cDNA clone, Mu-319, which contained the entire coding sequence for the cytoplasmic form of murine gelsolin was isolated using a polyclonal antibody. Gelsolin was detected in several cell lines but was not detectable in three undifferentiated embryonal carcinoma cell lines. Levels of gelsolin mRNA increased 10-fold during the differentiation of the murine embryonal carcinoma cell line, PC-13. Differentiation of PC-13 was accompanied by changes in cell shape, from small indistinct cells to large flat cells. The accumulation of gelsolin mRNA in PC-13 cells began 12-24 h after addition of the differentiation inducing agents. In comparison, 2-5A-dependent RNase activity showed a 40-fold increase beginning after 24 to 36 h and c-fos mRNA were shown to increase about 9 fold beginning 36 to 60 h after induction of differentiation. The levels of gelsolin per se, as determined by immunoreactivity were also shown to increase with differentiation of PC-13 cells. These results suggest that gelsolin may play a role in the restructuring of actin filaments which accompanies the dramatic changes in cell shape during differentiation. PMID- 2546952 TI - Expression and characterization of mutant forms of the type I regulatory subunit of cAMP-dependent protein kinase. The effect of defective cAMP binding on holoenzyme activation. AB - The mouse wild type and four mutant regulatory type I (RI) subunits were expressed in Escherichia coli and subjected to kinetic analyses. The defective RI subunits had point mutations in either cAMP-binding site A (G200/E), site B (G324/D, R332/H), or in both binding sites. In addition, a truncated form of RI which lacked the entire cAMP-binding site B was generated. All of the mutant RI subunits which bound [3H]cAMP demonstrated more rapid rates of cAMP dissociation compared to the wild type RI subunit. Dissociation profiles showed only a single dissociation component, suggesting that a single nonmutated binding site was functional. The mutant RI subunits associated with purified native catalytic subunit to form chromatographically separable holoenzyme complexes in which catalytic activity was suppressed. Each of these holoenzymes could be activated but showed varying degrees of cAMP responsiveness with apparent Ka values ranging from 40 nM to greater than 5 microM. The extent to which the mutated cAMP-binding sites were defective was also shown by the resistance of the respective holoenzymes to activation by cAMP analogs selective for the mutated binding sites. Kinetic results support the conclusions that 1) Gly-200 of cAMP-binding site A and Gly-324 or Arg-332 of site B are essential to normal conformation and function, 2) activation of type I cAMP-dependent protein kinase requires that only one of the cAMP-binding sites be functional, 3) mutational inactivation of site B (slow exchange) has a much more drastic effect than that of site A on increasing the Ka of the holoenzyme for cAMP, as well as in altering the rate of cAMP dissociation from the remaining site of the free RI subunit. The strong dependence of one cAMP-binding site on the integrity of the other site suggests a tight association between the two sites. PMID- 2546953 TI - Identification and characterization of the ligand binding subunit of a kainic acid receptor using monoclonal antibodies and peptide mapping. AB - Monoclonal antibodies (mAb) and a polyclonal antiserum were produced against a kainic acid receptor (KAR) purified from frog brain. Several of the mAb and the antiserum immunoprecipitated [3H]kainic acid binding activity from solubilized preparations of frog brain and labeled a group of proteins on immunoblots that migrated at Mr = 48,000. These results confirm that the ligand binding subunit of the frog brain KAR is contained in the Mr = 48,000 proteins. Immunoblots from different frog tissues demonstrated that the antibody reactivity was highly concentrated in the frog nervous system with no detectable immunoreactivity observed in non-neuronal tissues. The purified KAR was radioiodinated and subjected to two-dimensional gel electrophoresis and autoradiography. A series of proteins was detected at Mr = 48,000 with isoelectric points from 5.5 to 6.3. The anti-KAR mAb and the antiserum reacted with the same group of proteins from frog whole brain after separation by two-dimensional gel electrophoresis. Peptide maps of the 125I-labeled KAR separated by two-dimensional gel electrophoresis demonstrated that the group of proteins clustered at Mr = 48,000 is homologous. mAb KAR-B1 reacted on immunoblots with a protein in rat brain with a Mr = 99,000. This protein comigrated with an unreduced form of the KAR in frog brain. It was present in rat cerebral cortex, hippocampus, and cerebellum but was not detected in thalamus, globus pallidus, or brain stem, nor was it detected in rat non neuronal tissues. The presence of the Mr = 99,000 immunoreactive polypeptide in discrete areas of rat brain suggests that this protein may be part of a mammalian KAR or a related receptor. PMID- 2546954 TI - Presynapsis and synapsis of DNA promoted by the STP alpha and single-stranded DNA binding proteins from Saccharomyces cerevisiae. AB - We previously purified an activity from meiotic cell extracts of Saccharomyces cerevisiae that promotes the transfer of a strand from a duplex linear DNA molecule to complementary circular single-stranded DNA, naming it Strand Transfer Protein alpha (STP alpha) (Sugino, A., Nitiss, J., and Resnick, M. A. (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 3683-3687). This activity requires no nucleotide cofactor but is stimulated more than 10-fold by the addition of yeast single-stranded DNA-binding proteins (ySSBs). In this paper, we describe the aggregation and strand transfer of double-stranded and single-stranded DNA promoted by STP alpha and ySSB. There is a good correlation between the aggregation induced by various DNA-binding proteins (ySSBs, DBPs and histone proteins) and the stimulation of STP alpha-mediated DNA strand transfer. This implies that the stimulation by ySSBs and other binding proteins is probably due to the condensation of single-stranded and double-stranded DNA substrates into coaggregates. Within these coaggregates there is a higher probability of pairing between homologous double-stranded and single-stranded DNA, favoring the initiation of strand transfer. The aggregation reaction is rapid and precedes any reactions related to DNA strand transfer. We propose that condensation into coaggregates is a presynaptic step in DNA strand transfer promoted by STP alpha and that pairing between homologous double- and single-stranded DNA (synapsis) occurs in these coaggregates. Synapsis promoted by STP alpha and ySSBs also occurs between covalently closed double-stranded DNA and single-stranded linear DNA as well as linear double-stranded and linear single-stranded DNAs in the absence of any nucleotide cofactors. PMID- 2546955 TI - The two proteins of the erythropoietin receptor are structurally similar. AB - The structure of the erythropoietin receptor has been identified in this laboratory as two proteins of 100 and 85 kDa by cross-linking 125I-erythropoietin (125I-EP) to the surface of erythroid cells purified from the spleens of mice infected with the anemia strain of Friend virus. This study investigates the relatedness of these two proteins and the possibility that these proteins are subunits of the functional receptor for EP. Other workers have claimed that the 100- and 85-kDa proteins are bridged by disulfide bonds. This most likely is an artifact due to the insolubility of the cross-linked membrane. Proteolytic digestion by the method of Cleveland (Cleveland, D. W., Fischer, S. G., Kirschner, M. W., and Laemmli, U. K. (1977) J. Biol. Chem. 252, 1102-1106) resulted in identical fragments from the 100- and 85-kDa proteins, which strongly suggests that the primary amino acid sequence of these two proteins is similar if not identical. Increasing the number of protease inhibitors during the preparation of membranes and the binding and cross-linking steps increased the ratio of 100-kDa protein labeled compared to the 85-kDa protein. Together these results suggest that the 85-kDa protein is derived by proteolytic cleavage of the 100-kDa receptor for EP. It is not clear whether the 100-kDa protein can bind EP in the absence of the 85-kDa protein. PMID- 2546956 TI - Tumor-promoting activity and cytotoxicity of 3,4,3',4'-tetrachlorobiphenyl on N nitrosomorpholine-induced murine liver foci. AB - Effects of 3,4,3',4'-tetrachlorobiphenyl (TCB) on glucose-6-phosphatase (G6Pase) altered hepatic foci of N-nitrosomorpholine (NNM)-treated B6C3F1 mice were investigated. TCB was chosen as a selective 3-methylcholanthrene-type inducer and tumor promoter. To initiate hepatocarcinogenesis, mice were treated with NNM (160 mg/l, in drinking water for 7 weeks), as in previous studies with the rat model. After a treatment-free interval of 22 weeks, TCB was administered (5 x 50 mg/kg, every 3 days), and liver foci were analysed 10 weeks after the start of TCB treatment. Unexpectedly, the number of G6Pase-negative and -positive foci per liver was markedly diminished following TCB treatment (to 32% and 57%, respectively). On the other hand, the mean volume of the remaining G6Pase-altered foci was enhanced, owing to an increase in the percentage of foci of large size (greater than 0.5 mm2). Throughout the experimental period of 39 weeks prolonged liver injury due to NNM and TCB treatment was demonstrated by histology and by elevated serum levels of glutamate-oxaloacetate transaminase. The results suggest that (in contrast to the rat system) TCB exhibited opposing effects on liver foci in the mouse model: (a) moderate tumor-promoting effects and (b) cytotoxic effects in NNM-injured liver, leading to decreased numbers of liver foci. PMID- 2546957 TI - Heterogeneous alterations of UDP-glucuronosyltransferases in mouse hepatic foci. AB - UDP-glucuronosyltransferase (UDPGT) was studied immunohistochemically in hepatic foci and nodules of N-nitrosomorpholine-treated mice. Serial sections were stained for glucose-6-phosphatase (G6Pase). It was found that a high percentage of G6Pase-negative liver foci and nodules were also UDPGT-negative (34%). In addition, G6Pase-negative foci without altered UDPGT phenotype (30%) and UDPGT negative foci without altered G6Pase phenotype (8%) were detected. G6Pase positive foci were also present (24%). Interestingly, most G6Pase-positive foci were UDPGT-positive (16%). Some G6Pase-positive lesions without altered UDPGT phenotype were also found (8%). The major phenotype observed in rat hepatocarcinogenesis models (UDPGT-positive/G6Pase-negative foci) was not detectable in the mouse model. These results demonstrate heterogeneous alterations of UDPGTs in mouse hepatic foci. They furthermore suggest marked differences between the mouse and the rat in the regulation of UDPGTs in similarly induced rat hepatic foci. PMID- 2546958 TI - Simultaneous quantitative high-performance liquid chromatographic determination of cyclic adenosine 3',5'-monophosphate and adenosine in mouse brain after microwave irradiation: application to isoprenaline effect in vivo. PMID- 2546959 TI - Immunoglobulins of patients with idiopathic Addison's disease block the in vitro action of adrenocorticotropin. AB - The presence of serum immunoglobulins (Ig) blocking ACTH-induced adrenal DNA synthesis and/or cortisol production was studied in 25 patients with idiopathic Addison's disease. For this purpose guinea pig adrenal segments kept in organ culture were exposed to ACTH and graded concentrations of patient IgG. After a 5 h culture period the cortisol present in the culture fluid was measured by RIA, and DNA synthesis in the adrenal cells was measured using Feulgen densitometry on frozen sections of the cultured adrenal segments. Addition of ACTH alone in concentrations of 0.1-10 pmol/L to the culture system stimulated in vitro cortisol secretion; the maximal stimulation was 63 +/- 35% (+/- SD; n = 5) at a concentration 0.1 pmol/L. ACTH also increased (in concentrations of 1 fmol/L to 1 pmol/L) the percentage of fasciculata cells in S-phase from 0-4% (nonstimulated) to 5-12%. IgG preparations from all but 2 of the 25 patients with idiopathic Addison's disease blocked these in vitro ACTH-induced adrenal responses in a dose dependent fashion. IgG from 2 patients with tuberculous adrenalitis, 1 patient with secondary adrenal insufficiency, and 7 normal subjects had no blocking activity. Among 5 non-Addisonian autoimmune endocrinopathy patients who had adrenal cytoplasmic autoantibodies, 4 had no ACTH-blocking IgGs. Two of 9 patients with miscellaneous adrenal disorders (Cushing's disease, pigmented adrenal micronodular dysplasia, and adrenal nodules) had ACTH-blocking activity. These results demonstrate the existence of IgGs blocking the in vitro effects of ACTH and suggest their involvement in the pathogenesis of idiopathic Addison's disease. PMID- 2546960 TI - Angiotensin II stimulates both inositol phosphate production and human placental lactogen release from human trophoblastic cells. AB - We studied the functional significance of the binding of angiotensin-II (AII) to human placentas. Human trophoblastic cell suspensions were prepared by trypsin digestion of minced tissue. Cell incubations with increasing doses of [125I](SAR1)AII, ranging from 0.01-2.5 nmol/L, were carried out for 20 min at 37 C. The results indicated the presence of specific low capacity [4300 +/- 1300 (+/ SE) sites/cell], high affinity (Kd = 0.38 +/- 0.06 nmol/L) binding sites for [125I](Sar1)AII. This binding was specific for AII analogs. When placental cells were preloaded with 40 microCi/mL [3H]myoinositol for 2 h at 37 C, AII stimulation resulted in a dose-dependent increase in inositol phosphate (InsP) production [EC50 = 1.4 +/- 0.4 (+/- SE) nmol/L], as measured by ion exchange chromatography. (Sar1)AII also stimulated InsP production, with an EC50 of 0.3 +/ 0.2 nmol/L. AII-stimulated production of InsP was completely blocked by the antagonist (Sar1,Ala8)AII. AII also stimulated human placental lactogen release from trophoblastic cells in a dose-dependent fashion. The EC50 was 18 +/- 9 pmol/L, and the stimulation was blocked by (Sar1,Ala8)AII, as found for AII stimulated InsP production. These results suggest that stimulation of human placental lactogen release by AII may be mediated by activation of phospholipase C, which, in turn, produces phosphoinositide breakdown. The results, therefore, provide evidence of a physiological role for the renin-angiotensin system within the human placenta. PMID- 2546961 TI - Normal structure of the gonadotropin-releasing hormone (GnRH) gene in patients with GnRH deficiency and idiopathic hypogonadotropic hypogonadism. AB - Idiopathic hypogonadotropic hypogonadism (IHH) is a genetic disorder in which a lack of hypothalamic GnRH secretion forms the basis for impaired gonadotropin release. Several lines of evidence, including an animal model of GnRH deficiency, suggest that an abnormality at the level of the GnRH gene might be responsible for the secretory defect in patients with IHH. We examined 13 patients (11 males and 2 females) with IHH documented by standard clinical and biochemical criteria, including apulsatile basal LH secretion which was reversed by administration of pulsatile GnRH. The structure of the GnRH gene in these patients was determined using Southern blot analyses. In every case EcoRI digestion revealed a 10 kilobase fragment including the entire GnRH gene, eliminating deletion of the gene as a cause of IHH. Multiple restriction enzymes (Xba I, BglII, BamHI, PstI, TaqI, RsaI, EcoRV, and BglI) and hybridization probes to exons 2 and 4 of the gene were used to search for polymorphisms or relatively small deletions or rearrangements in the GnRH gene. Polymorphisms were not identified, and deletions or rearrangements involving more than 200 basepairs were excluded. We conclude that a major rearrangement of the GnRH gene is not a common basis for IHH in humans. PMID- 2546963 TI - Opioids stimulate growth hormone (GH) release in man independently of GH releasing hormone. AB - The Met-enkephalin analog DAMME [D-Ala2,MePhe4-Met-enkephalin-(o)-o1, FK 33-824] can stimulate GH secretion in man. In this study we investigated the effects of the guanyl derivative of DAMME (G-DAMME) on the serum GH response to an analog of GHRH in normal men. GHRH-(1-29)NH2 and G-DAMME each induced a rise in serum GH, and the increase was greater when both were given together. Since the GHRH-(1 29)NH2 dose (100 micrograms) used was a maximally stimulatory one, these results suggest that the enhancing effect of G-DAMME on GHRH-induced GH release may be mediated through inhibition of somatostatin release. PMID- 2546962 TI - Acromegaloid patients with type A insulin resistance: parallel defects in insulin and insulin-like growth factor-I receptors and biological responses in cultured fibroblasts. AB - A subset of patients with the syndrome of acanthosis nigricans and insulin resistance type A is characterized by acromegaloid features in addition to hyperinsulinemia, hyperandrogenemia, and an inherent defect in insulin receptor function. It has been proposed that the acromegaloid features result from the interaction of insulin at concentrations encountered in vivo, with a functionally intact insulin-like growth factor-I (IGF-I) receptor closely related to the insulin receptor. We investigated this possibility by examining binding and hormone-stimulated [14C]glucose uptake, [3H]thymidine uptake, and receptor autophosphorylation by both insulin and IGF-I in cultured fibroblasts from two affected patients. In comparison to normal fibroblasts, [125I]insulin binding, insulin-stimulated [14C]glucose, and [3H]thymidine uptake, and insulin-stimulated autophosphorylation were each reduced by approximately 50-60% of the absolute values in controls. In contrast to expectation, each of these apparent defects in insulin binding and action were mirrored by a parallel decrease in IGF-I binding and action. Thus, [125I]IGF binding was approximately 50%, IGF-I stimulated [3H]thymidine uptake was approximately 40% and 60% of the control value, and IGF I-stimulated receptor autophosphorylation was reduced by 40%. Incubation of fibroblasts with insulin at 25 ng/mL reduced subsequent binding of [125I]IGF-I by approximately 20% and did not enhance maximal stimulation of [3H]thymidine incorporation. We conclude that in some patients with acanthosis nigricans and acromegaloid features, IGF-I receptors of cultured fibroblasts may share the inherent defects of insulin receptor function. These in vitro data do not explain the acromegaloid features observed in vivo, suggesting that acromegaloid features are mediated by other mechanisms. PMID- 2546964 TI - Effect of administration of corticotropin-releasing hormone and glucocorticoid on arginine vasopressin response to osmotic stimulus in normal subjects and patients with hypocorticotropinism without overt diabetes insipidus. AB - We examined the effect of CRH administration on the response of plasma arginine vasopressin (AVP) induced by an osmotic stimulus in six normal subjects and five patients with hypocorticotropinism without overt diabetes insipidus (four patients with Sheehan's syndrome and one with idiopathic pituitary dwarfism with ACTH deficiency). Hypertonic saline infusion (855 mmol/L saline solutions at a rate of 205 mumol/kg.min for 10 min) increased plasma AVP 5.7-fold (P less than 0.01) in normal subjects and 2.4-fold (P less than 0.05) in the patients. CRH administration significantly augmented the plasma AVP response to the osmotic stimulus in the normal subjects, but not in the patients with hypocorticotropinism. CRH administration alone did not influence plasma AVP. These findings suggest that a central CRH-related mechanism(s) was at least partly involved in the augmentation of AVP release. Based on the relatively low plasma AVP response to the osmotic stimulus in patients and their lower plasma AVP levels and higher plasma osmolality under basal conditions, we suggest that patients with hypocorticotropinism have partial diabetes insipidus, in which impairment of central CRH action might be, at least in part, involved. The response of plasma AVP to the osmotic stimulus was attenuated significantly when the patients were given cortisol. Since basal PRA, plasma aldosterone, plasma osmolality, hematocrit, body weight, mean blood pressure, and heart rate were similar with and without cortisol administration, this effect of cortisol may have been due to central suppression of the AVP response to the osmotic stimulus. PMID- 2546965 TI - The neuroendocrine effects of interleukin-2 treatment. AB - Observations of neuropsychiatric changes in patients receiving interleukin-2 (IL 2) led us to examine the effects of IL-2 administration on the stress-related hormones, beta-endorphin, ACTH, cortisol, and CRH. We evaluated 30 cancer patients who received immunotherapy with IL-2 or IL-2 plus lymphokine-activated killer (LAK) cells. Blood samples were taken immediately before and 4 and 8 h after infusion of IL-2 or IL-2 plus LAK cells. IL-2 stimulated increased hormone levels 4 h after infusion compared with those before therapy and with basal levels in normal volunteers at the following magnitudes: beta-endorphin, 10-fold; ACTH, 20-fold; and cortisol, 2-fold. The effect of IL-2 was not altered in patients also receiving LAK cells. An effect of treatment course was noted, with higher stimulated values seen 4 h after IL-2 in the second treatment course compared with those after the first course [change (delta) in beta-endorphin, 101 vs. 11 fmol/mL; delta ACTH, 138 vs. 8 pmol/L; delta cortisol, 414 vs. 218 nmol/L]. We conclude that IL-2 treatment induces the release of neuroendocrine hormones and that a significant increase in hormonal stimulation occurs upon reexposure to IL-2. PMID- 2546966 TI - Interferon-gamma inhibits thyrotropin-induced thyroidal peroxidase gene expression in cultured human thyrocytes. AB - We have previously demonstrated that interferon-gamma (IFN-gamma) induced HLA-DR antigen and also inhibited thyrotropin (TSH)-induced triiodothyronine (T3) and thyroglobulin (Tg) secretion from cultured human thyrocytes. In order to further clarify the inhibitory effect of IFN-gamma on TSH-stimulated thyroid hormone secretion, we have examined human thyroid peroxidase (TPO) gene expression. Thyrocytes dispersed from Graves' thyroid tissues were incubated with TSH with or without IFN-gamma. Total RNA was extracted, separated and hybridized with 32P labelled human TPO cDNA. Thyrocytes expressed four TPO mRNA transcripts (4.0, 3.2, 2.1 and 1.7kb, respectively), all of which were stimulated by TSH. IFN-gamma inhibited TSH-stimulated TPO mRNA in a dose dependent manner (0.01-10U/mL). 1 U/mL IFN-gamma caused maximal suppression of TSH-stimulated TPO mRNA levels to basal levels. IFN-gamma also inhibited 8-bromo-cyclic AMP-stimulated TPO mRNA levels. In contrast, the gamma-actin mRNA hybridization signal was not altered in control or treated cells. These results demonstrate that IFN-gamma directly inhibits TSH-stimulated TPO gene expression and provide further support for a role of IFN-gamma as a local modulator of thyroid hormone synthesis. PMID- 2546967 TI - [A clinical evaluation of the versatility of various tumor markers in diagnosing the primary carcinoma of the lung]. AB - In this study, the author has evaluated the diagnostic versatility of the neuron specific enolase (NSE), carcinoembryonic antigen (CEA), tissue polypeptide antigen (TPA), and serum antigens (KA 32, KA 93) which are detected by anti-human lung carcinoma monoclonal antibodies (KM 32, KM 93) in patients before initiating any treatment. The positive rates of the serum NSE, CEA, TPA, KA 32 and KA 93 in all patients suffering from lung carcinoma were 31.4% (32/102), 52.8% (56/106), 63.3% (62/98), 52% (13/25) and 20% (24/120) respectively. Serum NSE was positive in 80.8% (21/26) of patients suffering from small cell type lung carcinoma (SCLC) and the mean value (32.7 +/- 25.4 ng/ml) was significantly higher than those of other varieties of lung carcinoma. The positive rate of serum CEA in adenocarcinoma (70.2%) was significantly higher than those of squamous cell carcinoma (22.2%). There was no significant statistical difference in positive rates of TPA in various histological types of lung carcinoma. The NSE and CEA were 44.0% (22/50) and 70.6% (36/51) in the stage IV disease and they appeared to reflect the progress and extent of the disease. The TPA tended to show a positive rate even at the initial stage of the disease, but, it was noteworthy that this disclosed a relatively high false positive rate of 54.2% (13/24). Moreover, determination of the serum NSE was performed chronologically. A lowered serum NSE value was confirmed in all cases which responded to the therapeutic attempts and unchanged values or even elevated values were noted in cases which showed no favourable response or rapid progression of the disease. It was also noteworthy that the serum NSE elevation was found 2-6 weeks prior to the clinical confirmation of the recurrence of the tumor in three patients suffering from SCLC. Based on these observations, it is suggested that the serum NSE may serve as a versatile tumor marker in monitoring the stage of disease, effectiveness of the therapeutic attempts and prediction of the possibility of the recurrence in SCLC. However, in view of the fact that some of the cases that obviously demonstrated clinical evidence of tumor recurrence failed to show elevation of the NSE, caution should be exercised in evaluating the alteration of the positive rates. The monoclonal antibody that works against human lung squamous cell carcinoma (KM 32) and antibody that works against human lung adenocarcinoma (KM 93) were isolated and purified.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2546968 TI - Evaluation of a commercial monoclonal antibody-based enzyme immunoassay for detection of adenovirus types 40 and 41 in stool specimens. AB - A commercial monoclonal antibody-based enzyme immunoassay (Adenoscreen; Mercia Diagnostics Ltd., Guildford, United Kingdom) for the detection of adenovirus types 40 and 41 in stool specimens was evaluated. Two assay modes were tested. In the first, 177 stool samples were screened for the presence of adenovirus type 40 or 41 (assay mode 1). Virus was detected in 79 of 82 specimens positive for adenovirus type 40 or 41 by a polyclonal antibody-based immune electron microscope test, giving a sensitivity of 96.3%. The enzyme immunoassay was negative in 91 of 95 stool samples which contained either other adenovirus serotypes or other viruses or were virus negative. The specificity was thus 95.8%. The positive and negative predictive values of this assay against immune electron microscopy were 95.2 and 96.8%, respectively, and the diagnostic accuracy was 96.0%. Viruses from the three false-negative enzyme immunoassay stool samples were verified as adenovirus type 40 or 41 by restriction enzyme analysis, monoclonal antibody-based immune electron microscopy, or both. Two of the three false-negative stool samples were subsequently concentrated by ultracentrifugation, and one of the two stool samples was then positive by enzyme immunoassay. The third false-negative virus was typed as adenovirus type 41 in the second (serotyping) enzyme immunoassay mode. The four enzyme immunoassay false-positive stool samples all contained other adenovirus serotypes (two were type 2, and two were type 5), but no cross-reactivity was seen with other strains of these serotypes and the results probably reflected simultaneous excretion of adenovirus type 40 or 41 with other adenovirus serotypes. In the second assay mode viruses from 15 stool samples were serotyped. The results by enzyme immunoassay (4 were type 40 and 11 were type 41) correlated completely with previous results from restriction endonuclease analyses. The commercial enzyme immunoassay system showed excellent sensitivity and specificity for the detection of adenovirus types 40 and 41 in stool specimens and will make an important contribution to the accurate diagnosis of adenovirus gastroenteritis. PMID- 2546969 TI - Clinical experience with cytomegalovirus isolation using conventional cell cultures and early antigen detection in centrifugation-enhanced shell vial cultures. AB - A total of 1,915 clinical samples was inoculated by low-speed centrifugation into shell vials (Bartels Immunodiagnostics, Bellvue, Wash.) containing cover slip monolayers of MRC-5 fibroblasts. At 1 and 2 days postinoculation, one cover slip was stained by an indirect immunofluorescence technique using a monoclonal antibody (Biotech Research Laboratories for Dupont, Billerica, Mass.) to cytomegalovirus (CMV) early antigen (EA). Clinical samples were also inoculated into three MRC-5 or MRHF cell cultures which were observed for 30 days for the appearance of a cytopathic effect (CPE). Of 157 CMV-positive samples, 92 (59%) were identified by centrifugation-enhanced EA (CE-EA) and 131 (83%) produced a CPE. CE-EA was less sensitive than CPE for all types of samples, although 17% of CMV-positive samples were detected by CE-EA alone. Evaluation of the CMV status of patients with CE-EA-positive-CPE-negative samples indicated that these samples likely represented true CMV-positive results. The average elapsed time between culture inoculation and identification of CMV decreased as follows when both CE EA and CPE, rather than CPE alone, were used: urines, 15 to 7 days; buffy coats, 18 to 9 days; lung samples, 13 to 8 days; throat samples, 18 to 7 days. Although CE-EA was less sensitive than 30-day cell culture, both CE-EA and CPE were identified as valuable in CMV detection, and neither could be discontinued without a decrease in the CMV isolation rate or an increase in the turnaround time. PMID- 2546970 TI - Development of an enzyme-linked immunosorbent assay for equine infectious anemia virus detection using recombinant Pr55gag. AB - To provide more sensitive and convenient methods for the detection of equine infectious anemia virus (EIAV), we developed an enzyme-linked immunosorbent assay (ELISA) employing the EIAV gag precursor (Pr55gag) produced by using recombinant DNA techniques. The antigenic reactivity of the recombinant EIAV Pr55gag was found to be equivalent to that of the virion p24gag and elicited high-titered antiserum in rabbits. When a large number of horse sera were analyzed for the presence of antibodies to EIAV by this ELISA, a radioimmunoassay for EIAV p15gag, or the standard agar gel immunodiffusion test, there was 98.7% concordance among the assays. By using the ELISA it was possible to specifically detect antibodies earlier after experimental infection of horses with EIAV than with the other two tests. A competition ELISA developed in order to detect EIAV gag antigens was found to be approximately 15 times more sensitive than the radioimmunoassay for EIAV p15gag. Antigens of other animal lentiviruses as well as those of the prototype oncovirus failed to compete in this assay. PMID- 2546971 TI - Detection of BK virus and JC virus in urine and brain tissue by the polymerase chain reaction. AB - DNAs of the human polyomaviruses BK virus (BKV) and JC virus (JCV) were amplified by the polymerase chain reaction (PCR) by using a single pair of 20-base oligonucleotide primers that were complementary to the same regions of both viruses. The sequences flanked by the primers were unique for each virus and could be differentiated by hybridization with 40-base, 32P-labeled oligonucleotide probes or by cleavage with BamHI. The DNA fragments resulting from amplification of BKV and JCV were 176 and 173 nucleotide pairs, respectively. The sensitivities of PCR for amplification of cloned BKV and JCV DNAs were 10 and 100 copies, respectively. Hybridization with the oligonucleotide probes was specific for each virus. A total of 57 urine samples from three groups of subjects were processed by DNA extraction or boiling and were tested by PCR. Urine samples collected from immunosuppressed patients (n = 11) and previously documented to be positive for BKV, JCV, or both were positive by PCR. Ten percent of urine samples from healthy adults (n = 30) that were previously negative for BKV and JCV were positive for one or both viruses by PCR. Urine samples (n = 16) from four seronegative bone marrow transplant recipients were uniformly negative for BKV. JCV was detected in deparaffinized brain tissue from a patient with progressive multifocal leukoencephalopathy. Specific diagnosis of virus in clinical specimens could be made within 1 day of receipt of the specimens. The PCR method is attractive for use in diagnosing polyomavirus infections because of its sensitivity, specificity, and rapid turnaround time. PMID- 2546972 TI - Polymerase chain reaction assay for detection of human cytomegalovirus. AB - Direct detection of human cytomegalovirus (HCMV) from clinical specimens was examined by using the polymerase chain reaction (PCR) for amplifying HCMV DNA. The efficiency of the amplification reaction was examined by using three different buffers and concentrations of deoxynucleotide triphosphates. The PCR assay was most efficient with a reaction mixture containing 17 mM ammonium sulfate, 67 mM Tris hydrochloride (pH 8.5), 7 mM MgCl2, 10 mM 2-mercaptoethanol, 170 micrograms of bovine serum albumin per ml, and each deoxynucleotide triphosphate at a final concentration of 1.5 mM. After 35 cycles of amplification, 0.15 fg of a plasmid containing the cloned target gene (corresponding to approximately six gene copies) was detected. The PCR assay correctly identified all of 24 clinical isolates of HCMV. Virus in urine specimens could be disrupted by heating at 93 degrees C for 30 min. The viral DNA was amplified directly from 5 microliters of preheated urine, with no further treatment before amplification. We tested the PCR assay on urine specimens from patients who had undergone renal transplantation that had been screened for the presence of HCMV by enzyme-linked immunosorbent assay, hybridization assay, and direct virus isolation. Specimens that were positive by one or more of these assays were screened by PCR. HCMV was consistently detected by PCR in all specimens that were positive by at least one other test. No cross-reactivity to other herpesviruses or MRC-5 cellular DNA was observed. PMID- 2546974 TI - Detection of rotavirus by hybridization with a nonradioactive synthetic DNA probe and comparison with commercial enzyme immunoassays and silver-stained polyacrylamide gels. AB - Three enzyme-linked immunosorbent assays (ELISAs) (rotavirus enzyme immunoassay [EIA; International Diagnostic Laboratories], Pathfinder [Kallestad Laboratories], and Rotaclone [Cambridge Bioscience, Inc.]) and hybridization of viral RNA with a nonradioactive, synthetic oligonucleotide DNA probe (SNAP; Molecular Biosystems, Inc.) were compared with silver-stained polyacrylamide gel electrophoresis (PAGE) of viral RNA for the detection of rotavirus in fecal specimens. Hybridization was performed following column purification of the viral RNA. Of 286 specimens analyzed by PAGE, SNAP, rotavirus EIA, Pathfinder, and Rotaclone, 88 were positive by PAGE. All 88 specimens were also positive by the other four assays. Nine specimens that were positive by one or more of the assays were positive by blocking ELISAs but were negative by PAGE. If these nine specimens were considered to be true positives, the final sensitivities and specificities were as follows: PAGE, 91 and 100%; SNAP, 94 and 97%; rotavirus EIA, 96 and 97%; Pathfinder, 100 and 94%; and Rotaclone, 96 and 97%, respectively. PMID- 2546973 TI - Comparison of monoclonal antibody time-resolved fluoroimmunoassay with monoclonal antibody capture-biotinylated detector enzyme immunoassay for respiratory syncytial virus and parainfluenza virus antigen detection. AB - An all-monoclonal antibody, time-resolved fluoroimmunoassay was compared with several enzyme immunoassays for the detection of respiratory syncytial virus and parainfluenza virus type 1, 2, and 3 antigens in clinical specimens. The most sensitive enzyme immunoassay for parainfluenza virus type 1 was an all-monoclonal antibody assay with biotin-labeled detector antibody and streptavidin-peroxidase conjugate, but for respiratory syncytial virus and parainfluenza virus types 2 and 3 the most sensitive assay was a polyclonal antibody assay with horse capture antibodies and bovine or rabbit detector antibodies with anti-species peroxidase. All tests were evaluated with nasopharyngeal aspirate specimens from respiratory illnesses and with cell culture harvests of multiple strains of each virus isolated over many years. The time-resolved fluoroimmunoassay detected respiratory syncytial virus antigen in 92% of the specimens positive by culture, which was a decidedly higher sensitivity than either the monoclonal or polyclonal antibody enzyme immunoassay format (62 and 76%, respectively). For the parainfluenza viruses the time-resolved fluoroimmunoassay detected type-specific antigen in 94 to 100% of culture-positive specimens and again was more sensitive than the all-monoclonal antibody enzyme immunoassays (75 to 89%) or all polyclonal antibody enzyme immunoassays (66 to 95%). Combined with results from a previously reported adenovirus time-resolved fluoroimmunoassay, these tests identified respiratory antigens in large numbers of clinical specimens. PMID- 2546975 TI - Detection of bluetongue virus antigens in Culicoides variipennis by enzyme immunoassay. AB - A solid-phase enzyme immunoassay (EIA) was developed to detect bluetongue (BT) virus antigens in infected cell cultures and in suspensions of infected Culicoides variipennis midges. The technique was equally sensitive for detecting the five U.S. BT virus serotypes (2, 10, 11, 13, and 17) in cell cultures. EIA reliably detected about 3.8 log10 median tissue culture infective doses per ml of BT virus in infected cell culture lysates. The EIA readily detected virus antigens in pools of midges infected with BT serotypes 2, 10, 11, 13, and 17 and contained 2.3 to 4.8 log10 median tissue culture infective doses per ml of BT virus. The technique was sensitive enough to detect a single infected midge in a pool with 99 noninfected midges. The EIA may be a sensitive and rapid alternative to virus isolation for surveillance of BT viruses in vector populations. PMID- 2546976 TI - Amplification techniques for detection of herpes simplex virus in neonatal and maternal genital specimens obtained at delivery. AB - Viral culture amplification methods, including centrifugation, use of a cell pretreatment medium containing dimethyl sulfoxide and dexamethasone (DEX medium), and three commercial enzyme immunoassays (EIAs), were evaluated for use with low titer, asymptomatic neonatal and obstetric samples obtained at delivery. The 4.5 h Ortho EIA was 59.7% sensitive at 16 h compared with 86.4% at 36 h in a spin amplified format (SAT-EIA). After 36 h with DEX medium, the SAT-EIA was 97.3% sensitive using the 2.5-h modified Ortho EIA and 69.9% sensitive using the Fairleigh Dickinson EIA. Herpes simplex virus (HSV) cytopathic effect was reported in tube culture 1 to 2 days earlier in 8.1 to 35.9% of cells with DEX medium. Overall, the SAT-EIA using the modified Ortho EIA and cells with DEX medium detected 147 of 153 (96.1%) HSV isolates from samples obtained predelivery and 93.1% of 29 HSV positives from samples obtained at labor and delivery, with specificities of 100 and 99.9%, respectively, at 36 h. PMID- 2546977 TI - Molecular epidemiology and restriction site mapping of adenovirus type 3 genome types. AB - One hundred thirty-eight strains of adenovirus type 3 isolated from patients in the United States, West Germany, and other regions between 1961 and 1984 were analyzed with six restriction endonucleases; 18 genome types were found. BglII was the most discriminative enzyme. Mapping of altered restriction sites was also performed for all six enzymes. The genome types D1 (like the prototype) and D3 prevailed among 45 and 47 strains, respectively. All genome types could be divided into two groups related to D1 or D3. Several clusters of infections by strains with the same genome type were observed. Only hints of differences were found in the pathogenicities of individual genome types. D1 strains were present in the United States and in Europe; group D3 prevailed almost exclusively in the United States. PMID- 2546978 TI - Serological evidence for variation in the incidence of herpesvirus infections in different species of apes. AB - Sera from captive lowland gorillas, chimpanzees, orangutans, and gibbons were screened by enzyme-linked immunosorbent assay (ELISA) for antibody to herpesviruses serologically related to human herpes simplex virus types 1 and 2 (HSV-1, HSV-2), a baboon virus (SA8), and a macaque herpesvirus (B virus). The incidence of herpesvirus antibodies varied considerably among the different species, gorillas having the highest incidence of seropositivity (65.4%) and orangutans the lowest. The virus specificity of positive sera was further analyzed by examining the kinetics of virus neutralization, competition of reactivity in ELISAs, and immunoblotting against HSV-1, HSV-2, SA8, and B virus antigens. Using these assays, the majority of positive gorilla sera (49 of 53, 92%) were determined to react in a manner identical to human HSV-1 immune sera. The remaining four positive gorilla sera reacted as HSV-2-positive sera. In contrast, the majority of positive chimpanzee sera (5 of 7, 71%) reacted as HSV-2 immune rather than HSV-1 immune. All positive sera from gibbon apes reacted as HSV-1 positive. No orangutan sera were identified which gave positive reactions by ELISAs to any of the four primate herpesviruses tested. Although four orangutan sera gave equivocal results against HSV-1 antigen, further analysis by immunoblotting could not confirm any specific reactivity with any of the primate herpesvirus antigens. Varied reactivity among individual animals with both SA8 and B virus proteins was observed, but none of the seropositive primates detected appeared to be infected with either of these simian viruses. Three gorilla sera had antigen recognition patterns slightly different from those of HSV-2-positive human and chimpanzee sera and another HSV-2-positive gorilla serum, raising the possibility that these animals harbor an indigenous virus related to HSV-2. PMID- 2546979 TI - Hepatitis A virus hemagglutination and a test for hemagglutination inhibition antibodies. AB - Like enteroviruses, hepatitis A virus (HAV) hemagglutinated various species of erythrocytes under similar conditions. HAV-specific antibodies in both acute- and convalescent-phase sera were found to inhibit hemagglutination. The HAV hemagglutination inhibition test can be used for diagnosis, epidemiological surveillance, and vaccine assessment. PMID- 2546980 TI - Serum immunoglobulin A response to Norwalk virus infection. AB - We describe the serum immunoglobulin A (IgA) antibody response to Norwalk virus infection in human volunteers and compare it with previously described IgM and total antibody responses. Whereas specific IgA and IgM peak within 2 weeks after onset of symptoms, titers of total blocking antibody continue to rise, implying mediation by IgG antibody. PMID- 2546982 TI - Dermal hematopoiesis in neonates: report of five cases. AB - We report five cases of dermal hematopoiesis in newborn infants. Dermal hematopoiesis, manifested clinically as a blueberry muffin eruption, has been associated with both intrauterine viral infections and congenital hematologic dyscrasias. Of our five patients, four of whom were boys, the cutaneous findings could be attributed to congenital infections in two instances. The infectious agents were cytomegalovirus and coxsackie virus B-2. Two infants had hematologic abnormalities, including twin transfusion syndrome and ABO blood group incompatibility. In our fifth patient a cause was not identified. PMID- 2546981 TI - Nerve growth factor receptor and choline acetyltransferase colocalization in neurons within the rat forebrain: response to fimbria-fornix transection. AB - Although it is well known that magnocellular cholinergic basal forebrain neurons are trophically responsive to nerve growth factor (NGF) and contain NGF receptors (NGFr), the exact distribution of forebrain NGFr-immunoreactive neurons and the degree to which cholinergic neurons are colocalized with them have remained in question. In this study we employed a very sensitive double-labelling method and examined in the same tissue section the distribution and cellular features of NGFr-positive and choline acetyltransferase (ChAT)-immunolabelled neurons within the rat basal forebrain. Throughout this region the majority of magnocellular basal forebrain neurons were immunoreactive for both NGFr and ChAT. However, a small percentage of neurons in the ventral portion of the vertical limb of the diagonal band of Broca were immunoreactive only for NGFr, whereas a larger population of magnocellular neurons in the substantia innominata exhibited only ChAT immunoreactivity. No NGFr-immunoreactive cells were found associated with ChAT-positive neurons in the striatum, neocortex, or hippocampus, and no single labelled NGFr-immunoreactive neurons were found outside the basal forebrain area, except for a large number of positive-labelled cells along the ventricular walls of the third ventricle. In addition to its function in maintaining the normal integrity of the basal forebrain and cholinergic, peripheral sympathetic, and neural-crest-derived sensory neurons, NGF may also have a role in the growth of these neurons after damage to the nervous system. To examine this postulate the hippocampus was denervated of its septal input and examined 8 weeks later. Two populations of neurons were found to have undergone collateral sprouting--namely, the midline magnocellular cholinergic neurons of the dorsal hippocampus and the sympathetic noradrenergic neurons of the superior cervical ganglion. Both of these neuronal populations also stained strongly for NGFr. In contrast, the small intrinsic cholinergic neurons of the hippocampus exhibited neither sprouting response nor staining for NGFr. In view of these results, we suggest that the differing sprouting responses demonstrated by these three neuronal populations may be due to their responsiveness to NGF, as indicated by the presence or absence of NGF receptors. PMID- 2546983 TI - Failure of sodium bicarbonate baths in the treatment of aquagenic pruritus. PMID- 2546984 TI - A diphosphoinositide kinase in rat mast cell granules. AB - Intact granules were isolated from sonicated purified rat serosal mast cells on a Percoll gradient. The granules were demonstrated to contain a diphosphoinositide kinase that catalyzes the formation of triphosphoinositide from diphosphoinositide. The enzyme requires adenosine triphosphate and Mg2+ or Mn2+ for activity. The Km for adenosine triphosphate is 3 mumol/L, and maximal response is observed at 20 mmol/L of Mg2+ or 1 mmol/L of Mn2+, respectively. Triphosphoinositide synthesis in the granules is dependent on the time and temperature of the incubations. Cyclic adenosine monophosphate, adenosine, adenosine diphosphate, and adenosine monophosphate decrease the enzyme activity. A comparison of the rate of phosphorylation of intact and broken membrane granules suggests that the phosphorylation occurs on the outer (cytoplasmic) surface of the granules. PMID- 2546985 TI - Leukotrienes, LTC4 and LTB4, in bronchoalveolar lavage in bronchial asthma and other respiratory diseases. AB - Leukotrienes (LTs) C4 and B4 are potent proinflammatory mediators with a wide variety of biologic activities, including smooth muscle contraction, mucus hypersecretion, and leukocyte activation, which may be of particular relevance to the pathology of asthma and other respiratory diseases. We measured the concentrations of LTC4 and LTB4 in bronchoalveolar lavage fluid from 16 atopic subjects with asthma (eight symptomatic and eight asymptomatic) and from 14 control subjects without asthma (six with hay fever and eight nonatopic). The amounts detected in symptomatic subjects with asthma were significantly higher than in control subjects (LTB4, 0.58 +/- 0.06 versus 0.36 +/- 0.05 pmol/ml, p less than 0.05; LTC4, 0.36 +/- 0.1 versus 0.12 +/- 0.02 pmol/ml, p less than 0.01). LTC4 and LTB4 were also measured in 17 patients: nine with interstitial lung disease of varying etiology (cryptogenic fibrosing alveolitis [CFA] or idiopathic pulmonary fibrosis), three with sarcoidosis, one with extrinsic allergic alveolitis, one with sulphonamide-induced pneumonia, and one patient with eosinophilic granuloma. The concentrations of LTB4 (but not LTC4) were significantly greater in patients with CFA compared with normal control subjects (0.69 +/- 0.3 versus 0.36 +/- 0.05 pmol/ml, p less than 0.01). There was a significant correlation (p less than 0.05) between the percentage of neutrophils and the concentration of LTB4 in the bronchoalveolar lavage fluid) of the group with interstitial lung disease as a whole. This study provides evidence for a role for LTs in the airways of subjects with day-to-day asthma and suggests that LTB4 may also be involved in the recruitment of granulocytes into the lung in patients with CFA. PMID- 2546986 TI - Alzheimer's dementia and binding to alpha 2 adrenoreceptors in platelets. AB - Seventy-five patients with probable Alzheimer's disease were screened for binding of alpha 2 receptors (A2R) to their platelet membranes; the results were compared with 51 age- and sex-matched controls. Receptor binding assays were performed using [3H] Yohimbine as the radioligand. The results showed a higher binding capacity in the demented population as compared to the control group (2.18 +/- 0.15 fmol/mg protein, as compared to 1.73 +/- 0.13, P less than 0.03). This increased binding to platelets in the demented patients was more prominent in demented females: 34% higher binding as compared with female controls (2.06 +/- 0.5 vs 1.54 +/- 0.04). The difference between demented and normal males was less (2.34 +/- 0.05 vs 1.88 +/- 0.05). The results indicate an involvement of the A2R system, either primarily or secondarily, in the disease process. Since there is an overlap between results from the patients with Alzheimer's disease and the normal subjects, A2R may serve as only a supportive marker for Alzheimer's disease. PMID- 2546987 TI - Medicare coverage in the nursing home. PMID- 2546988 TI - 5-Hydroxytryptamine produces presynaptic facilitation of cholinergic transmission in rabbit parasympathetic ganglia. AB - Intracellular recordings were made from neurons of rabbit vesical pelvic (parasympathetic) ganglia (VPG). Application of 5-hydroxytryptamine (5-HT, 0.3-30 microM) produced an initial depression followed by a long-lasting facilitation of the fast excitatory postsynaptic potential (e.p.s.p.) evoked by stimulation of the pelvic preganglionic nerve. The facilitation of nicotinic transmission lasted for 30-120 min, even when 5-HT was removed from the superfusing solution. 5-HT (0.3-30 microM) did not change the depolarization induced by a direct application of acetylcholine (ACh) to the VPG neurons pretreated with 1 microM atropine. 5-HT also caused an initial depression followed by an increase in the quantal content of the fast e.p.s.p. It is, therefore, suggested that diphasic effect of 5-HT on the nicotinic transmission is due mainly to a modulation of the ACh-release from presynaptic nerve terminals. Methysergide (5 microM), mianserin (5-30 microM) and ICS 205-930 (100-300 nM) did not antagonize the presynaptic actions of 5-HT on the nicotinic transmission, suggesting that the presynaptic 5-HT receptor may belong to a class of 5-HT1 subtypes. Spiperone (1 microM), a selective 5-HT1A antagonist, blocked the 5-HT-induced inhibition of the fast e.p.s.p. Under the effect of spiperone, the facilitation appeared soon after application of 5-HT. The facilitation of the fast e.p.s.p. may be mediated through a 5-HT1B or 5-HT1C subtype. Lowering temperature of the external solution eliminated the 5-HT induced facilitation of the nicotinic transmission. Forskolin produced a presynaptic facilitation of the fast e.p.s.p., without producing an initial depression. 3-Isobutyl-1-methylxanthine (10 microM) potentiated the facilitatory action of 5-HT. Bath-application of dibutyryl cyclic adenosine monophosphate (cAMP) (1-6 mM) and 8-bromo-cyclic AMP (2-5 mM) mimicked the effect of 5-HT in producing the facilitation of the fast e.p.s.p.s. All data presented are consistent with the hypothesis that 5-HT, acting on presynaptic 5-HT1 receptors, causes a facilitation in the release of ACh from preganglionic nerve terminals possibly mediated through an activation of adenylate cyclase. PMID- 2546989 TI - Small cell carcinoma of the colon. A case report and literature review. AB - We report a patient with two synchronous foci of small cell carcinoma in the rectosigmoid region, metastatic to the liver and paraaortic lymph nodes, and review forty-five other reported cases. Most frequent in the rectosigmoid and cecal regions, these tumors are highly aggressive, and distant metastasis at presentation is almost always the rule. The mean survival is approximately 6 months, and 1 year survival only 15%. A tentative management strategy is proposed. PMID- 2546990 TI - Chronic Epstein-Barr virus infection: a cause of granulomatous hepatitis? AB - A 37-year-old man had fever, weight loss, malaise, right upper quadrant pain, lymphadenopathy, and a twofold elevation of serum alkaline phosphatase, transaminases, and gamma glutamyl transpeptidase. Granulomas were found on liver biopsy after 2 1/2 months of illness. Treatment with isoniazid and rifampin for 2 months did not lead to improvement in fever or symptoms, but prednisone caused a prompt resolution. Positive IgM antibodies to the viral capsule antigen followed by marked elevation of the IgG fraction suggest chronic Epstein-Barr virus infection to be the etiology. The patient was asymptomatic without treatment 14 months after onset of the illness. PMID- 2546991 TI - Polarized distribution of Na+,K+-ATPase in giant cells elicited in vivo and in vitro. AB - Giant cell formation was analyzed to determine whether it results in the high level of Na+,K+-ATPase expression that characterizes multinucleated cells such as osteoclasts. Giant cells and fusing alveolar macrophages were subjected to morphological, immunological, and biochemical studies. Both subunits of the Na+,K+-ATPase were found to be present on the plasma membrane of giant cells. Their localization was restricted to the non-adherent domain of the cell surface. Dynamic studies of giant cell differentiation demonstrated that on culture and/or multinucleation, an increase in sodium pump alpha-subunit synthesis occurred and led to a high level of expression of Na pumps. Conversely, the adherent plasma membrane of giant cells was enriched in a lysosomal membrane antigen. This study demonstrates that culture and/or multinucleation induces a significant increase in the expression of sodium pumps. The polarized distribution of these pumps and of a lysosomal component suggests that fusing macrophages undergo biochemical and morphological alterations which prepare them for a new and specialized function in chronic inflammatory reactions. Giant cells may offer a suitable model system to study the differentiation of other related multinucleated cells, such as osteoclasts. PMID- 2546992 TI - Determination of human antibodies to specific antigens of cytomegalovirus using an antigen capture immunoassay. AB - The antigen capture immunoassay which is described herein is based on the binding of specific antigens of cytomegalovirus (CMV) by monoclonal antibodies bound to a solid phase. The specificity of the binding was demonstrated by the analysis of antigens labelled with [35S]methionine and captured by the bound monoclonal antibodies. These specific antigens are recognized in turn by specific anti cytomegalovirus antibodies in human sera. The immunoassay permits quantitation of these specific anti-cytomegalovirus antibodies and should facilitate both qualitative and quantitative comparisons of the antibodies against specific CMV antigens in different individuals. PMID- 2546993 TI - Phosphoinositides from human foetal brain during development. AB - An attempt has been made in the present study to look for the activity of the enzymes which catalyse the breakdown of phosphorylated metabolites both in acidic and alkaline pH from human foetal brain, in different gestational ages during development in 18 foetal samples. It was revealed that phosphatases were distributed throughout the brain and were operative from the onset of development. There was a tendency of increment of specific activity of enzymes with respect to the advancement of gestational period. This observation should be accounted as a prerequisite criteria for understanding phosphate meatabolism in human foetal organs primarily associated with glycolytic and gluconeogenic pathways. PMID- 2546995 TI - Endothelial barrier function. AB - The endothelial barrier in all organ beds allows the free exchange of water, but is restrictive to varying degrees to the transport of solutes such as albumin. For example, in the brain microvessels, the endothelial barrier restricts the transport of protein, whereas in fenestrated and continuous endothelial cells of the renal and lung endothelial cells, the endothelial barrier is semipermeable. The endothelial monolayer demonstrates selectivity, i.e., the permeation of molecules is inversely related to the molecular weight. Although the "pore" theory has been used to describe the transport across the endothelial barrier, the transport of solutes is also dependent on the charge of solutes and the endothelial cell, and the ability of the solute to bind to or be taken up by endothelial cells. Receptor-mediated trancytosis of albumin may contribute to albumin transport in addition to transport by paracellular pathways (i.e., through a so-called pore). Water permeability across the endothelium is determined by the interaction of albumin with glycocalyx and interstitial components of the endothelium (the "fiber matrix"). Ambient concentration of albumin serves to lower endothelial hydraulic conductivity. Increased endothelial permeability to solutes and water in inflammatory states is dependent on the shape and configuration of endothelial cells as determined by alterations in cytoskeletal elements, such as f-actin, and as regulated by intracellular second messengers such as free cytosolic calcium. PMID- 2546994 TI - Dispase, a neutral protease from Bacillus polymyxa, is a powerful fibronectinase and type IV collagenase. AB - Dispase, a neutral protease isolated from culture filtrates of Bacillus polymyxa, has proven to be a rapid, effective, but gentle agent for separating intact epidermis from the dermis and intact epithelial sheets in culture from the substratum. In both cases it effects separation by cleaving the basement membrane zone region while preserving the viability of the epithelial cells. Because it is not known what or where in the basement membrane zone Dispase cleaves, we set up studies to define its substrate specificity. Using purified basement membrane components and sodium dodecyl sulfate-polyacrylamide gel electrophoresis we show that Dispase cleaves fibronectin and type IV collagen, but not laminin, type V collagen, serum albumin, or transferrin. The action of Dispase on collagen appears to be selective for type IV collagen in that several stable degradation products are formed, whereas the enzyme degrades type I collagen only minimally. In newborn human skin, as seen by electron microscopy, Dispase removes the lamina densa, rich in type IV collagen, but preserves the anchoring fibrils (structures known to contain type VII collagen) and the epidermal cells. Because its action is so selective, it suggests that Dispase can serve as a powerful tool for dissecting epithelial-mesenchymal interactions. PMID- 2546996 TI - Lymphocyte recognition of psoriatic endothelium: evidence for a tissue-specific receptor/ligand interaction. AB - Epidermotropic lymphocytes are an essential cellular component of the skin associated lymphoid tissues (SALT). Dermal lymphocytic infiltrates are also characteristics of inflammatory dermatoses, such as psoriasis, and may be involved in the pathogenesis of the disease, although the mechanisms by which lymphocytes are recruited into these sites are not known. We have used an in vitro lymphocyte adherence assay to test the hypothesis that specialized endothelial cells are present in inflamed skin, and are capable of supporting lymphocyte adherence and promoting lymphocyte emigration. In this assay, we assessed the binding of lymphocytes overlaid onto frozen sections of normal and psoriatic skin. Peripheral blood mononuclear cells (PBMC) from patients, from healthy volunteers, and from rat thoracic duct bound specifically to dermal endothelia in psoriatic plaques, in steroid-resistant areas of plaques, but not in uninvolved skin or skin from healthy individuals. Analysis of the binding properties of lymphocyte subsets revealed preferential adherence by CD4+ T cells as compared with CD8+ T cells and to B cells. Interestingly, PBMC from patients undergoing ultraviolet light therapy failed to adhere to autologous skin or to lesion-containing skin sections from untreated patients. Additional studies indicate that the lymphocyte-endothelial interaction is an energy- and calcium dependent process and involves surface glycoprotein and carbohydrate moieties, requirements similar to those found in specific lymphocyte interactions with high endothelial venules in lymph nodes during the homing process. Pretreatment of lymphocytes with antibodies directed against homing receptors mediating migration into lymph nodes and into gut-associated lymphoid tissues, however, did not interfere with lymphocyte adherence to psoriatic endothelium. In contrast, anti lymphocyte function associated antigen (LFA)-1 antibody partially inhibited lymphocyte binding to chronic plaques. We conclude that a tissue-specific receptor/ligand interaction independent of LFA-1 directs lymphocyte emigration from the dermal microvasculature into the psoriatic dermis and that LFA-1 plays only an accessory role in the adhesion process. PMID- 2546997 TI - Pharmacokinetics and toxicity of inhaled human interferon-alpha in patients with lung cancer. AB - Eight patients with advanced, previously treated non-small cell lung cancer inhaled natural leukocyte interferon-alpha (IFN-alpha) from a dosimeter-equipped jet nebulizer. Single doses of IFN ranged from 1 x 10(6) to 120 x 10(6) IU. Serum IFN was undetectable after single doses of 1 x 10(6) to 18 x 10(6) IU, but 60 x 10(6) IU resulted in measurable levels of circulating IFN in 3 out of 6 patients. All 5 patients who inhaled 120 x 10(6) IU had between 11 and 35 IU of IFN per milliliter of serum for at least 12 h. No systemic or local side effects were observed after 1 x 10(6) to 18 x 10(6) IU, but doses of 60 x 10(6) to 120 x 10(6) IU resulted in temperature rise, headache, and malaise. All symptoms started within 3-6 h, reached their peak by 8-10 h, and lasted until 12-24 h after inhalation. A decrease (greater than 20%) in peak expiratory flow following inhalation was temporarily found in 2 patients. We conclude that IFN, given by inhalation, penetrates into the blood stream, thus causing systemic side effects similar to those described after systemic IFN administration and, in addition, occasionally reversible airflow obstruction. PMID- 2546998 TI - Effect of amiloride on the compensatory renal growth that follows uninephrectomy in mice. AB - The increase in contralateral kidney mass that follows the removal of one kidney is caused primarily by the hypertrophy and, to a lesser extent, the hyperplasia of tubule cells. Amiloride inhibits the proliferation of cells in tissue culture environments and diminishes compensatory hyperplasia of liver cells after partial hepatectomy. In the current study, we determined the extent to which amiloride diminished the compensatory increase in contralateral kidney weight and protein content after uninephrectomy. Caworth Farms mice, 3 to 4 months of age, received amiloride by daily intraperitoneal injection for 7 days before the left kidney was removed and for an additional 4 days after nephrectomy. Four days after uninephrectomy, compensatory hypertrophy was quantified by the increase in weight and protein content of the right kidney in comparison with the left (R - L divided by L x 100; [weight] +24.7% +/- 2.9% and [protein] +20.6% +/- 4.4%). Treatment with amiloride reversibly reduced the compensatory increase in kidney weight and protein content (dose required to achieve 50% inhibition, 11.4 and 2.4 mg/kg/day, respectively). Hexamethylene amiloride, an analog with weak affinity for conductive Na+ channels and high affinity for Na+-H+ exchangers, had no effect on compensatory hypertrophy. Spironolactone, a natriuretic and antikaliuretic agent of potency similar to that of amiloride, but with a different cellular mechanism of action, had no effect on compensatory hypertrophy. We conclude that amiloride reversibly blunts the compensatory renal hypertrophy after uninephrectomy in mice by mechanisms that appear to be separate from the drug's effects to block Na+-H+ exchange and transepithelial Na+ and K+ transport. PMID- 2546999 TI - Evaluation of reduced activity galactose-1-phosphate uridyl transferase by combined radioisotopic assay and high-resolution isoelectric focusing. AB - Fifty-nine patients referred for testing of galactose-1-phosphate uridyl transferase (GALT) activity were evaluated by a combination of radioisotopic assay and an improved high-resolution isoelectric focusing (IEF) method for determination of individual genotypes. The majority of evaluations (43 of 59) were undertaken because of an abnormal fluorometric GALT screening assay conducted as part of a state newborn galactosemia screening program or as a diagnostic test for an infant with one or more clinical signs of galactosemia. Only 12 of the 43 patients with an abnormal fluorometric screening test were found to have less than 3% GALT activity by quantitative assay, no GALT-staining by IEF, and a clinical history consistent with classical galactosemia. The majority of the remainder (24 of 43) were found to be heterozygotes for galactosemia, the Duarte variant, or both. IEF was found to be a fast and accurate technique for assignment of GALT genotypes in conjunction with quantitative enzymatic assay. PMID- 2547000 TI - Effects of amiloride on the endolymphatic sac. AB - The effect of amiloride on the murine endolymphatic sac was investigated. The amiloride caused collapse of the lateral intercellular spaces in the endolymphatic sac epithelium and a subsequent mild endolymphatic hydrops. These changes indicated a decreased absorption of endolymph in the endolymphatic sac. Amiloride is known to inhibit the transcellular fluid transport without inducing any changes in the paracellular fluid transport. It is therefore suggested that amiloride specially inhibits the fluid and ion exchange in the apical portion of the epithelial cells resulting a decrease in transcellular fluid transport across the endolymphatic sac epithelium. The transcellular fluid transport seems to be one of the main mechanisms in the endolymphatic sac fluid exchange system. PMID- 2547001 TI - Proton magnetic resonance spectroscopy of plasma from patients with dyslipoproteinemia: identification of factors governing methyl and methylene proton line widths. AB - The line width of the proton magnetic resonance spectrum (MRS) of the composite methylene and methyl resonances of plasma has been reported as a marker for the presence of malignancy. In this study, the contribution of very low density (VLDL), low density (LDL), and high density lipoproteins (HDL) to the MRS line width was determined. This was achieved by measuring the MRS line widths for the plasma from patients with primary disorders of lipoprotein metabolism and from normal individuals. A negative correlation between plasma trigylceride levels and the average line width was observed and this was confirmed in normal plasma to which pure VLDL was added. Also, computer simulations were employed to demonstrate how the line width varies in such complex mixtures of lipoproteins. We demonstrate that the line width is governed by the relative contribution of VLDL and HDL to the composite line shape. This is particularly important when the shoulder from the HDL line lies near the half-height of the VLDL line. As changes in VLDL/HDL ratio occur in patients with malignancy, we propose that this is the basis of the narrowed MRS lines observed in the proposed test for malignancy. However, any individual with elevated VLDL will be false positive in this test. PMID- 2547002 TI - Effects of selective opioid-receptor blockade on the hypothalamo-pituitary adrenocortical responses to surgical trauma in the rat. AB - Opioid substances have been shown to stimulate and depress the secretion of ACTH in the rat. Their opposing actions appear to be mediated in part by specific receptors in the hypothalamus which influence the secretion of corticotrophin releasing factor (CRF). In an attempt to determine the physiological role of these receptor systems, experiments were carried out in which the plasma ACTH and serum corticosterone concentrations were determined before and after stress in rats treated s.c. with selective antagonists of mu-(naloxone, naltrexone), delta (ICI 174864) and kappa-(MR2266) opioid receptors. Neither naloxone (25-100 micrograms/100 g) nor naltrexone (50 micrograms/100 g) influenced the resting plasma ACTH or serum corticosterone concentrations. However, both inhibited (P less than 0.01) the secretion of the two hormones elicited normally by surgical stress (laparotomy under ether anaesthesia). ICI 174864 (30-100 micrograms/100 g) also had little effect on resting hypothalamo-pituitary-adrenocortical (HPA) activity but, at the highest dose, it caused a small (P less than 0.05) potentiation of the response to surgery. In contrast, MR2266 (150-300 micrograms/100 g) produced marked activation of the HPA system and not only stimulated the resting secretion of ACTH and corticosterone but also potentiated and prolonged the HPA response to stress. The results suggest that mu- and kappa opioid receptors mediate opposing actions of endogenous opioid peptides, both of which may be physiologically important in the regulation of CRF release. PMID- 2547003 TI - Immunofluorescent localization of type II insulin-like growth factor receptor in rat liver and hepatoma cells. AB - We have used an immunofluorescent technique to localize type II insulin-like growth factor (IGF) receptors in rat liver, rat hepatocytes and three rat hepatoma cell lines (HTC, H-35 and 5123) using a polyclonal antibody (C-1) raised to purified rat liver type II IGF receptor. Specificity of the antiserum was confirmed by Western blotting of microsomal membranes prepared from hepatocytes and hepatoma cells which showed a single class of receptor in all cells, of Mr approximately 210,000 for hepatocytes, HTC and H-35 cells and approximately 220,000 for 5123 cells, on non-reduced, 4-15% polyacrylamide gradient gels. The specificity of the immunofluorescent technique was also verified by abolition of labelling after preincubation of antiserum with purified type II IGF receptor. Rat liver cryosections contained areas of juxtanuclear labelling in hepatocytes, consistent with the presence of type II IGF receptor in the Golgi region. Brightest immunofluorescence was seen in sections from fetal and neonatal rats with adult rat hepatocytes staining brightly only around central veins. Areas of labelling were also seen in connective tissue surrounding larger veins. Cultured adult rat hepatocytes and rat hepatoma cell lines also showed bright areas of juxtanuclear nuclear immunofluorescence, with HTC and H-35 cells staining more than 5123 and adult hepatocytes. Fetal rat hepatocytes in culture also labelled very brightly both in a juxtanuclear location and in small clusters over the cell, possibly on the cell surface. These observations indicate that type II IGF receptors are located predominantly on intracellular membranes and are most abundant in rapidly growing cells and tissues (such as fetal liver and hepatoma cells). PMID- 2547004 TI - Dopaminergic regulation of pituitary gonadotrophin-releasing hormone receptor activity in the goldfish (Carassius auratus). AB - In goldfish, dopamine acts as an endogenous inhibitor of basal and gonadotrophin releasing hormone (GnRH)-stimulated gonadotrophin release. The purpose of the present study was to investigate the effects of dopamine on the pituitary GnRH receptors in vivo and in vitro in goldfish. The goldfish pituitary contains two classes of GnRH-binding sites, a high-affinity/low-capacity site and low affinity/high-capacity site. Injection of domperidone, a dopamine antagonist, resulted in a dose- and time-related increase in capacity of both the high- and low-affinity GnRH-binding sites; apomorphine, a dopamine agonist, completely reversed this effect. The effects on GnRH receptor capacity correlated very closely with changes in serum gonadotrophin concentrations. Domperidone was generally without effect on GnRH-binding affinity; however, a small but significant decrease in affinity was observed for the low-affinity binding site at 18 h after injection of the highest dose of domperidone used (40 mumol/kg body weight). Treatment with apomorphine of goldfish pituitary fragments in a perifusion system caused a decrease in the capacity of both the high- and low affinity GnRH-binding sites without affecting binding affinity; domperidone reversed this effect. It is concluded that the dopaminergic inhibition of basal and GnRH-stimulated gonadotrophin release in goldfish might, in part, be the result of a down-regulation of the pituitary GnRH receptors; this effect of dopamine can be achieved by a direct action at the pituitary level. PMID- 2547005 TI - Adrenal mast cells modulate vascular and secretory responses in the intact adrenal gland of the rat. AB - Mast cells were identified in the rat adrenal gland, located in the walls of arterioles at the point at which they penetrate the connective tissue capsule. The mast cell products, histamine and serotonin, both caused dose-dependent increases in rates of perfusion medium flow and steroid secretion in the isolated, perfused rat adrenal gland in situ. Compound 48-80, a mast cell degranulator, caused a significant increase in perfusion medium flow rate and steroid secretion by the in-situ perfused rat adrenal. Administration of disodium cromoglycate, a mast cell stabilizer, before administration of ACTH(1-24) virtually abolished the normal flow rate increment and significantly attenuated the corticosterone secretory response to ACTH(1-24). These observations strongly suggest that adrenal mast cells modulate both vascular and secretory responses in the intact adrenal gland of the rat. PMID- 2547006 TI - Characterization of the steroidogenic responsiveness and ultrastructure of purified zona fasciculata/reticularis cells from bovine adrenal cortex before and after primary culture. AB - Analysis by electron microscopy indicated that after 3 days of primary culture, purified bovine adrenal zonal fasciculata/reticularis (ZF/ZR) cells showed improved integrity of their ultrastructure, with an increased density of lipid droplets and smooth endoplasmic reticulum. The basal cortisol output was significantly (P less than 0.05) greater on day 3 of culture than for the freshly isolated cells in six out of seven experiments. Similarly, in six experiments with ACTH (1 nmol/l) and five experiments with angiotensin II (10 nmol/l), the stimulated cortisol secretion was significantly (P less than 0.01 for all 11 experiments) higher on day 3 of culture than in freshly isolated cells. No significant increase in cortisol secretion above basal was observed with noradrenaline at any concentration in the freshly isolated cells, whereas a dose dependent increase in cortisol secretion was observed on day 3 of culture in all of four experiments. These findings were supported by cyclic (c) AMP output measured in one such experiment. Thus the basal cAMP output and that stimulated by ACTH (1 nmol/l) were significantly higher after culture (P less than 0.001, n = five wells for basal comparison; P less than 0.05, n = three wells for ACTH at 1 nmol/l). In agreement with the cortisol results, cAMP production was unaffected by any concentration of noradrenaline in the freshly isolated cells, whereas a dose-dependent rise was found after culture. Angiotensin II at all concentrations had no effect on cAMP production in freshly isolated or cultured cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547007 TI - Effect of maternal hypothyroxinaemia during fetal life on the calmodulin regulated phosphatase activity in the brain of the adult progeny in the rat. AB - Calmodulin-regulated phosphatase activity was measured in the brain of 2-month old rats born from hypothyroid and normal dams, using a fluorometric enzyme assay developed for this purpose. Calmodulin content was measured in the same brain regions by radioimmunoassay. Significant differences between groups in weight and protein content, basal phosphatase and calmodulin-regulated phosphatase activity were found. The brain region most affected was the cerebellum, where basal and calmodulin-regulated phosphatase activities, and protein content were increased. The data point towards a lasting effect of maternal hypothyroxinaemia on the brain function of the progeny. PMID- 2547008 TI - Possible role of histones in the organization of rat liver thyroid hormone receptors in chromatin. AB - The effects of histone subfractions on rat liver thyroid hormone receptor-DNA interaction were examined using an in-vitro DNA-cellulose binding assay. H1 histones bound to DNA showed reversible and potent inhibition of receptor-DNA binding without affecting receptor hormone binding. Poly-lysine, bovine serum albumin, ovalbumin and cytochrome c did not alter receptor-DNA binding. H1 histone subfractions (calf thymus lysine-rich histone (CTL)-1, CTL-2 and CTL-3) showed potent inhibition of receptor-DNA binding indistinguishable from each other. The quantity of H1 histone subfractions bound to DNA was the same. Although each subfraction has different functional properties, inhibition of receptor-DNA binding was a common feature of all the H1 histone subfractions, which is important for the non-random distribution of the receptor in chromatin. Binding of the receptor to core histones was investigated; it was found to bind to core histones more potently than to other proteins (H1 histone, ovalbumin and cytochrome c). Among core histone subfractions, H4 histone bound to the receptor most potently and is the candidate to be one of the acceptor sites of the receptor in chromatin. PMID- 2547009 TI - Effect of short-term starvation on reproductive hormone gene expression, secretion and receptor levels in male rats. AB - The effects of 4-6 days of food deprivation on the pituitary-testicular function of adult male rats were studied. Fasting decreased body weights on average by 23% (P less than 0.01) and those of seminal vesicles by 55% (P less than 0.01) in 4 days. No consistent changes were found in testicular and ventral prostate weights. The pituitary levels of gonadotrophin-releasing hormone (GnRH) receptors decreased by 50% (P less than 0.01). Serum and pituitary levels of LH, FSH and prolactin decreased by 25-50% (P less than 0.01 for all). Testicular and serum levels of testosterone decreased by 70-80%, testicular LH receptors by 26%, those of prolactin by 50% (P less than 0.01 for all), but those of FSH remained unaffected. Acute (2 h) stimulation by a GnRH agonist (buserelin, 10 micrograms/kg i.m.) resulted in similar LH, FSH and testosterone responses in the fasted and control animals, and human chorionic gonadotrophin (hCG) stimulation (30 IU/kg i.m.) in similar increases in testosterone. A 42% decrease was found in pituitary content of mRNA of the common alpha subunit (P less than 0.05), but the mRNAs of the LH- and FSH-beta chains and prolactin were unaffected by fasting for 4 days. When the same mRNAs were measured after 6 days of fasting, the decrease of the mRNA of FSH-beta also became significant (50%, P less than 0.01). In contrast, the mRNA of LH-beta was increased twofold (P less than 0.01) at this time and serum LH levels were similar in control and starved animals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547010 TI - Inhibition of arginine vasopressin-stimulated Na+ K+ ATPase activity by difluoromethyl ornithine in the rat renal medulla. AB - Arginine vasopressin (AVP) stimulates Na+ K+ ATPase and ornithine decarboxylase (ODC) activity in the rat medullary thick ascending limb. The effect of difluoromethyl ornithine (DFMO), a specific inhibitor of ODC activity, on AVP stimulated Na+ K+ ATPase activity was evaluated using a cytochemical bioassay. Peaks in Na+ K+ ATPase activity in cultured rat renal segments which occurred after tissue had been exposed to 1 fmol AVP/l were completely inhibited by the addition of 20 mmol DFMO/l to the culture medium containing AVP. The addition of 20 mmol DFMO/l to the culture medium containing AVP in the concentration range 0.001-10 fmol/l inhibited completely the stimulation of Na+ K+ ATPase activity by AVP. The response of Na+ K+ ATPase to increasing doses of ATP (10-40 g polypeptide/l) was not influenced by the addition of 20 mmol DFMO/l to the culture medium containing AVP, suggesting that the prevention of AVP-stimulated Na+ K+ ATPase activity by DFMO was not due to a direct effect on the enzyme. PMID- 2547011 TI - Effect of ouabain on cellular free calcium and cellular cyclic AMP production in response to arginine vasopressin in rat renal papillary collecting tubule cells in culture. AB - The effect of extracellular calcium (Ca2+) on the cellular action of arginine vasopressin (AVP) was examined using an Na+, K+-ATPase inhibitor in rat renal papillary collecting tubule cells in culture. The pretreatment of cells with ouabain enhanced basal and AVP-induced cAMP production in a dose-dependent manner. The augmentation by ouabain of cellular cAMP production in response to AVP was totally abolished by co-treatment with cobalt, lanthanum, verapamil or Ca2+-free medium containing 1 mmol EGTA/l, each blocking cellular Ca2+ uptake by different mechanisms. Two other findings indicated that ouabain directly stimulated cellular Ca2+ mobilization; namely, that ouabain significantly increased 45Ca2+ influx and cellular free Ca2+ concentration [( Ca2+]i) determined by Fura-2 fluorescence. The ouabain-induced increase in [Ca2+]i was completely blocked by either cobalt or Ca2+-free medium containing 1 mmol EGTA/l. AVP at 0.1 mumol/l increased [Ca2+]i to 177.1 +/- 26.2 nmol/l from 92.2 +/- 8.0 nmol/l (P less than 0.01) in renal papillary collecting tubule cells, and ouabain significantly enhanced the AVP-induced increase in [Ca2+]i. The increase of cellular free Ca2+ induced by ouabain probably binds to calmodulin to form an active complex of Ca2+-calmodulin in the cell, since two chemically dissimilar antagonists of calmodulin attenuated the enhancement by ouabain of cAMP production in response to AVP. These results therefore indicate that ouabain increases cellular Ca2+ uptake and enhances AVP-induced cellular free Ca2+ mobilization and its own second messenger cAMP production in renal papillary collecting tubule cells, and that extracellular Ca2+ is an important source for ouabain-mobilized cellular Ca2+. PMID- 2547012 TI - Actions of desacetyl-alpha-melanocyte-stimulating hormone on human adrenocortical cells. AB - The responses of human adrenocortical cells to stimulation by ACTH(1-24), desacetyl-alpha-MSH, alpha-MSH and angiotensin II amide have been compared. Both desacetyl-alpha-MSH, thought to be the major form of the peptide in the human pituitary and in circulating plasma, and alpha-MSH caused a significant stimulation of aldosterone, corticosterone and cortisol secretion. Significant stimulation of the production of these steroids was obtained with desacetyl-alpha MSH at a concentration of 1 nmol/l, while the response to alpha-MSH was considerably less sensitive, with a minimum effective concentration of 0.1 mumol/l. These values compared with minimum effective concentrations of 1 pmol/l for ACTH and 0.1 mumol/l for angiotensin II amide. Although cell types were not separated, it is possible to conclude that none of the peptides showed any specificity for the zona glomerulosa, and in each case the same minimum effective concentration of peptide was required for both aldosterone and cortisol secretion. Yields of steroid obtained under conditions of maximal stimulation by ACTH(1-24), alpha-MSH and desacetyl-alpha-MSH were at least three to five times the basal output of aldosterone, four to eight times that for corticosterone and more than seven to sixteen times that for cortisol. Angiotensin II amide was a relatively poor stimulant with maximal stimulation only 1.5 x basal. In these experiments the minimum effective concentration for desacetyl-alpha-MSH (1 nmol/l) was close to the circulating concentration of desacetyl-alpha-MSH (0.3 nmol/l) in man, and it is thus possible that this peptide may have a physiological role in the control of adrenocortical function. PMID- 2547013 TI - Simultaneous neuroinfection with cytomegalovirus and herpes simplex virus in an immunocompetent adult. AB - An immunocompetent adult developed Guillain-Barre Syndrome and encephalitis simultaneously. Serological and cerebrospinal fluid analyses provided evidence of both cytomegalovirus and herpes simplex virus infections. The patient recovered with only moderate sequelae following acyclovir treatment. PMID- 2547014 TI - Enzyme linked immunosorbent assays (ELISA) for the quantitative determination of human leukocyte collagenase and gelatinase. AB - A competitive and a sandwich enzyme linked immunosorbent assay (ELISA) were developed for human leukocyte collagenase and gelatinase. The competitive assay could detect 0.5 ng collagenase and 0.05 ng gelatinase. The detection limit of the sandwich ELISA was 0.05 ng for collagenase and 0.02 ng for gelatinase. No cross reactivity between human leukocyte collagenase and gelatinase was detected. The sandwich ELISA was used to determine plasma levels of these enzymes. The 90% range for collagenase was between 0 and 50 micrograms/l; the 90% range for gelatinase was between 27 and 94 micrograms/l. PMID- 2547015 TI - Influence of lithium carbonate on the thyrotropin receptor in vitro. AB - Thyroid tissues excised from 5 patients with Graves' disease were used to study the influence of lithium carbonate on the thyrotropin (TSH) receptor in vitro by cAMP generation assay and TSH binding to its receptor. Lithium carbonate was able to decrease the cAMP production of the crude thyroid plasma membranes stimulated by TSH at concentrations of 6mM and higher. However, lithium carbonate had no effect on 5'-guanylylimidodiphosphate-induced cAMP production in thyroid plasma membranes up to concentrations of 10mM. Lithium carbonate was able to decrease the binding of 125I-TSH to crude thyroid plasma membranes at concentrations of 6mM and higher. It decreased the binding of 125I-TSH to solubilized TSH receptor at concentrations of 6mM and higher. These findings suggest that lithium carbonate can influence the TSH receptor in vitro and may be one of the action mechanisms of lithium on the thyroid gland. PMID- 2547016 TI - Animal model of ultraviolet-radiation-induced recurrent herpes simplex virus infection. AB - The development of a model of induced recurrent herpes simplex virus disease would facilitate studies of the mechanism(s) responsible for reactivation of latent virus. The guinea pig model of genital herpes is characterized by a self limited primary infection, the establishment of a latent infection in sensory ganglia, and the subsequent development of spontaneous recurrent genital infections. This study reports that exposure of latently infected female guinea pigs to ultraviolet radiation results in the induction of recurrent genital herpes simplex virus infections in approximately 60% of animals. Furthermore, there was a dose-response relationship between ultraviolet irradiation and ability to induce recurrent disease. This model should prove useful in further investigations of the pathophysiology of herpes simplex infections. PMID- 2547018 TI - Typing of human papillomaviruses in cervical intraepithelial neoplasia grade 3 biopsies from Cape Town. AB - DNA from 98 cervical intraepithelial neoplasia grade 3 (CIN 3) biopsies was screened, using the Southern blotting hybridization technique, for human papillomavirus (HPV) types 6, 11, 16, 18, 31, 33, and 35. HPV 16 was detected in 16 biopsies (16%), HPV 33 in eight biopsies (8%), HPV 31 in two biopsies (2%), and HPV 18 in one biopsy (1%). One of the biopsies contained both HPV 31 and 18. Six biopsies (6%) contained an HPV type very similar but not identical to HPV 16, and 35 biopsies were positive for HPV, but the Pst 1 restriction fragments were distinct from any of the HPV types used as probes. HPV was not detected in 32% (32/98) of the biopsies screened. PMID- 2547017 TI - Characterization of cytomegalovirus isolates recovered during repeated infection in renal transplant recipients. AB - The sources of cytomegalovirus (CMV) infection in seropositive renal transplant recipients include reactivation of latent endogenous virus in the recipient, reactivation of latent virus in donated kidney, or both. If infection occurs by either source, viruses isolated from the same recipient should be the same strain, whereas if it occurs by both sources, those from the same recipient should be different. In this study, we followed prospectively 25 seropositive recipients who predominantly received a kidney from a seropositive donor to determine whether CMV isolates recovered repeatedly from them are the same or different. During an average 10 month follow-up period, six (24%) patients had excreted the virus more than twice at any site and/or at different times. Restriction enzyme analysis of viral DNA prepared by the Hirt procedure revealed that three or four isolates obtained from each of five patients were same, whereas six isolates from one patient included three different strains. Five of six patients had clinical symptoms at the times when CMV was recovered. Three patients with acute rejection and one patient with hepatitis had been infected with one single strain, and all were successfully treated. One patient with fever and acute rejection had been infected with three different strains, and he failed to recover his renal function. These results suggest that most CMV infections in seropositive recipients may be caused by one single strain. However, multiple infections with different strains can also occur. Such infections are associated with more severe clinical disease. PMID- 2547019 TI - Comparison of three techniques for detection of respiratory viruses in nasopharyngeal aspirates from children with lower acute respiratory infections. AB - A comparison of immunofluorescence (IF), enzyme-linked immunosorbent assay (ELISA), and isolation in tissue culture (TC) for detection of respiratory viruses was performed on 496 nasopharyngeal aspirates from children under 5 years of age with lower acute respiratory infections who were receiving attention at three hospitals in Buenos Aires, Argentina. All samples were tested by the three methods for respiratory syncytial virus (RSV), influenza A and B, adenovirus, and parainfluenza 1 and 3. Viral diagnosis was made in 167 samples (33.7%); of these, 124 (74.3%) were isolated in TC, whereas 120 (71.8%) were detected by ELISA and 127 (76%) by IF. RSV was detected in 121 samples, mainly by ELISA and IF. The sensitivity and specificity of each rapid technique as compared with isolation in TC were similar, reaching 98% and 92%, respectively. When ELISA was compared with IF, the sensitivity was 95%, and the specificity was 98%. Adenovirus was detected in 18 patients by TC. For this virus, rapid techniques sensitivity as compared with TC was low (almost 22%). Parainfluenza 3 was readily detected by IF and TC; influenza A, B and parainfluenza 1 were detected in few samples; and tissue culture proved more efficient than rapid techniques. The results indicate that both rapid techniques are good tools for the detection of most respiratory viruses except for adenovirus, for which TC cannot be omitted. PMID- 2547020 TI - Molecular identification of a novel human rotavirus in relation to subgroup and electropherotype of genomic RNA. AB - A total of 41 stool rotavirus specimens collected from children with acute diarrhea at four different locations in Akita Prefecture, Japan, during the peak of the winter diarrhea epidemic in 1988 were analyzed by polyacrylamide gel electrophoresis of viral RNA in conjunction with subgrouping assay. We found that a single strain predominated, with cocirculating strains with less common electropherotypes at a given location, and that two different strains could predominate at geographically close but different locations even during a very limited time of the epidemic season. Furthermore, we isolated a human rotavirus strain (AU125) that was similar to the AU-1 strain in that it possessed a long RNA pattern yet belonged to subgroup I. Genetic analysis by RNA-RNA hybridization assay indicated that the AU125 strain was distinct from two previously identified human rotavirus gene groups (genogroups) represented by the Wa strain (subgroup II with long RNA electropherotype) and the DS-1 strain (subgroup I with short RNA electropherotype), but was very closely related to the AU-1 strain. These data suggest that the genetic diversity of human rotaviruses may be more extensive than was previously thought. PMID- 2547021 TI - Infection by enteric adenoviruses, rotaviruses, and other agents in a rural African environment. AB - From February 1985 to January 1986, 432 stool samples, 310 from rural African children with diarrhea and 122 from controls, were analysed for the presence of enteric viruses known to be associated with diarrhea. Group A rotavirus ELISA indicated 12.9% positivity among patients and 2.5% positivity among controls. Only 23 of the 43 rotavirus ELISA-positive stools were also positive by electron microscopy. Nine children, three of whom were controls, were found to be shedding coronavirus-like particles, detected by electron microscopy. Stools from all but one of the nine children had been taken within 1 month of each other. Dot-blot hybridization tests for the presence of Ad40 or Ad41 DNA revealed 44 positive stools, 41 of which were from patients (13.2% positivity). Only three of the Ad40 or Ad41-positive stools by DNA hybridization were positive by electron microscopy, and only these three strains could be grown in semipermissive Chang conjunctival cells and their identity checked by restriction enzyme analysis. Further attempts to rescue the other strains using a helper virus failed, but nine of the stools proved positive by ELISA using a subgroup F-specific monoclonal antibody. On the basis of the DNA hybridization results alone, subgroup F adenoviruses were encountered as frequently as rotavirus in the study and were significantly associated with diarrhea, although the viability and intactness of virus particles by the time of laboratory analysis appeared to be very low. PMID- 2547022 TI - HBV-DNA hybridization in hepatocellular carcinoma associated with alcohol in Japan. AB - To clarify the role of the hepatitis B virus (HBV) in hepatocellular carcinoma (HCC) associated with alcohol consumption, HBV-DNA in the liver of 19 patients with HCC were investigated. HBV-DNA was examined by Southern blot hybridization. HBV-DNA was integrated into tumor cells from five out of six (83%) patients with HCC associated with HBs antigen (HBsAg)-positive post-hepatitic liver cirrhosis (LC), but this was not related to the history of alcohol intake. In 13 HCC patients of HBsAg-negative alcoholic LC, HBV-DNA integration was not detected in any patient. These findings suggest that HBV does not play a major role in the pathogenesis of HCC in HBsAg-negative alcoholics in Japan. PMID- 2547024 TI - Detection of neutralizing IgM antibodies in the diagnosis of enterovirus infections. AB - Data are presented of IgM detection by a neutralization test used routinely in 1,062 patients. Antigens isolated during the period of investigation were EV4, EV7, EV11, EV18, EV21, EV24, EV33, CA9, CB2, CB4, and CB5. No difference was observed in the distribution of IgM-positive sera according to age and sex. Total antibodies are at higher titres when IgM antibodies are present. Polytypic IgM responses are not frequent (less than 10%). The frequency of the IgM-positive sera for a given serotype correlated with the frequency of isolates for the serotype except for CA9. Other than for babies under age 6 months, IgM detection is more frequent than is isolation. The susceptibility of the elderly and the frequency of IgM-positive sera among adults over age 40 years suggests possible underestimation of enterovirus infections in adults. The duration of IgM remains a major question. PMID- 2547023 TI - Regulation by dimethylsulfoxide, insulin, and corticosteroids of hepatitis B virus replication in a transfected human hepatoma cell line. AB - A human hepatoblastoma cell clone E4 was obtained by transfection of HepG2 cells with a plasmid DNA containing four tandem copies of hepatitis B virus (HBV) genome. Analysis of both intracellular and extracellular viral DNA revealed that this clone exhibited the main steps of the replication process previously found in normal hepatocyte primary cultures experimentally infected in vitro. Indeed, relaxed-circular, covalently closed circular, and single-stranded forms of viral DNA were identified in the cells together with complete virions and immature cores in the medium. Furthermore, the ability of these secreted particles to infect normal human hepatocyte cultures was established. These E4 cells were used to evaluate the effect of various soluble factors on HBV replication. Corticosteroids and, to a greater extent, dimethylsulfoxide (DMSO) increased intracellular viral DNA, whereas insulin reduced it dramatically. Parallel changes in the amounts of viral DNA secreted in the medium were observed. Measurement of the albumin secretion rate indicated that cellular and viral activities could be regulated, at least in part, in a coordinated manner. PMID- 2547025 TI - Relationship between electrocortical activity and beta-adrenergic receptor function in the rat after chronic desimipramine treatment. AB - The aim of this study was to investigate the effects of chronic administration of desimipramine (DMI) after 2, 7 or 20 mg/kg per day, administered by osmotic minipumps, on electrocortical activity and beta-adrenergic receptors in rat brain. Rats receiving DMI chronically show a dose- and time-dependent increase of electrocortical activity above 15 Hz as well as a dose- and time-dependent decrease below 15 Hz. Already after 3 days of treatment a clear effect on the electrocorticogram (ECoG) was seen. The maximal change in the ECoG was reached at the end of the study, after 24 days of treatment. After acute treatment (20 and 45 minutes after 2, 4 or 10 mg/kg i.p.) with DMI, a decrease of electrocortical activity is seen above 15 Hz. Thus the effect of acute DMI treatment on the ECoG is different from that of chronic treatment. In the same group of rats the effect of chronic DMI treatment on the beta-adrenergic receptor number was determined 24 hours after the last ECoG recording. The number of beta-adrenergic receptors was dose dependently reduced in the DMI-treated rats as determined by [3H] dihydroalprenolol binding. There was no change in affinity (KD) of the ligand for the beta-receptor. This finding was corroborated by a decrease in the functional activity of the beta-adrenergic receptors, as determined by isoprenaline stimulated efflux of cyclic-AMP in cortex slices. These data indicate that chronic treatment with DMI, resulting in a down-regulation of the cortical beta adrenergic system, is paralleled by pronounced effects on the ECoG of rats. The different ECoG profiles after chronic DMI treatment compared with acute treatment suggest that adaptive changes in the electrical brain activity continually develop during the chronic treatment with this antidepressant drug. PMID- 2547026 TI - Caffeine withdrawal affects central adenosine receptors but not benzodiazepine receptors. AB - The effects of chronic caffeine administration on both adenosine and benzodiazepine receptors were studied in mouse brain membranes. Animals were fed on a diet enriched with caffeine (600 mg/kg diet) for 15 days and sacrificed 2, 4, 8 and 15 days after withdrawal. Compared with controls fed on a regular diet, animals receiving a caffeine-enriched diet showed an increase in the number of brain adenosine receptors labeled with [3H]-DPX in both the cerebellum and forebrain regions. This up-regulation was still significant 15 days after withdrawal in the cerebellum but not in the forebrain, where the number of adenosine receptors returned to control levels within 8 days following withdrawal. Benzodiazepine receptors labeled by [3H]-B-CCE were not influenced by chronic caffeine diet or withdrawal. PMID- 2547027 TI - Neuropathology in movement disorders. AB - This review concentrates on the definition and classification of degenerative movement disorders in which Parkinsonian symptoms are often prominent. The pathological spectrum and clinical manifestations of Lewy body disease are described, and associations with Alzheimer's disease and motor neuron disease are explored. A classification of pallidonigral degenerations is based on clinical features, distribution of pathology, and morphological abnormalities; some of these patients have mild nigral degeneration and no Parkinsonian features. Many other juvenile and familial Parkinsonian cases are not included among the pallidonigral degenerations. Most of these latter syndromes have been organised into preliminary groups, in particular, autosomal dominant dystonia-Parkinson syndrome, juvenile Parkinsonian disorder and autosomal dominant Lewy body disease. PMID- 2547028 TI - Cisplatin versus cisplatin plus etoposide in the treatment of advanced non-small cell lung cancer. Lung Cancer Working Party, Belgium. AB - We conducted a randomized study comparing the survival after treatment with cisplatin (120 mg/m2) or cisplatin plus etoposide (100 mg/m2 on days 1, 2, and 3) in 162 evaluable patients with advanced non-small-cell lung cancer (NSCLC). No statistically significant difference in survival was detected; the median survival was 26 and 22 weeks, respectively, for patients receiving cisplatin and for those receiving cisplatin plus etoposide. The objective response rate was 19% for cisplatin and 26% for the combination; the corresponding response rates were 17% and 43% in patients with limited disease. No significant differences were detected between the two study arms as far as toxicity was concerned, except for alopecia and granulocytopenia, which occurred more frequently in patients treated with cisplatin plus etoposide. PMID- 2547029 TI - Systemic therapy for small-cell lung cancer: old themes replayed, new ones awaited. PMID- 2547031 TI - The blood-brain barrier and response of C.N.S. metastases to chemotherapy. AB - A case demonstrating a differential effect of chemotherapy on a pineal metastasis and parenchymal cerebral metastases is described. At presentation, extensive metastatic small cell carcinoma of the lung was present and CT scanning showed an apparently solitary metastasis in the pineal. The clinical course and serial CT scans showed significant improvement of the pineal tumor and simultaneous development of multiple intra-cerebral metastases. This case confirms that the pineal gland is excluded from the blood-brain barrier, and indicates the clinical importance of the effect of the blood-brain barrier in the responsiveness of CNS metastases to chemotherapy. PMID- 2547030 TI - A randomized trial of anticoagulation with warfarin and of alternating chemotherapy in extensive small-cell lung cancer by the Cancer and Leukemia Group B. AB - The Cancer and Leukemia Group B (CALGB) conducted a prospective randomized trial to evaluate the role of warfarin and alternating chemotherapy in extensive small cell lung cancer (SCCL). After stratification for sex and performance status, patients were randomly assigned to receive chemotherapy with methotrexate, doxorubicin (Adriamycin; Adria Laboratories, Columbus, OH), cyclophosphamide, and lomustine (CCNU) (MACC), or MACC plus warfarin (MACC + W), or mitomycin, etoposide, cisplatin, and hexamethylmelamine alternating with MACC (MEPH/MACC). Warfarin was given continuously to maintain a prothrombin time of one and one half to twice the control values. A total of 328 patients were enrolled, and 294 were evaluable. There was a statistically significant advantage in objective response rates (complete [CR] and partial responses [PR], respectively) for MACC + W (17% and 50%) as compared with MACC alone (8% and 43%) or MEPH/MACC (10% and 38%) (P = .012). Both failure-free survival (P = .054 Wilcoxon test) and overall survival (P = .098 Wilcoxon test) were higher on MACC + W (median, 6.6 months and 9.3 months, respectively), as compared with MACC (5.0 months and 7.9 months) and MEPH/MACC (5.0 months and 7.9 months). Toxicity was comparable among the three arms, except for increased hemorrhagic events on MACC + W, which were life threatening in four patients (4%), and lethal in two others (2%). These data support the role of warfarin in the treatment of SCCL, but do not establish its mechanism of action. Warfarin deserves further studies in SCCL, particularly in patients with limited disease. PMID- 2547032 TI - Intra-arterial cisplatin for the treatment of malignant gliomas. AB - Cisplatin (DDP) is a chemotherapeutic agent that has shown efficacy against primary CNS malignancies. Intra-arterial (IA) administration of DDP to patients with brain tumors should produce higher peak levels of drug than intravenous (IV) administration of an identical dose and reduce systemic toxicity. Twelve patients with malignant glioma were entered into the study. All had failed irradiation, 11 had failed IA BCNU. Each patient received IA DDP, 58-100 mg/m2, into the internal carotid artery at four to six week intervals. One of 12 patients had a partial response of 6 months. The remaining 11 patients had progressive disease or severe complications. Toxicity included seizures in four patients, weakness and/or aphasia in four patients, coma in two patients, and visual deterioration in two patients. IA DDP has very limited efficacy in patients with malignant gliomas after failure of nitrosoureas and is associated with an unacceptable level of toxicity. IA DDP may be more effective when used as initial chemotherapy of malignant gliomas. PMID- 2547034 TI - Hypoxic changes in hippocampal neurons. AB - 1. Reversible effects of brief periods of anoxia (replacing 95% O2-5% CO2 with 95% N2-5% CO2 for 2-4 min) were studied in CA1 neurons in hippocampal slices (from Sprague-Dawley rats), kept in an interface-type chamber at 33.5 degree. 2. The predominant voltage change during anoxia (N2) was a hyperpolarization, accompanied by a marked fall in resistance and excitability; synaptic potentials were also depressed, especially inhibitory postsynaptic potentials (IPSPs). 3. In voltage-current (V-I) plots, the N2-evoked hypolarization had a reversal potential below -90mV, even when recording with 2 M KCl electrodes and after substituting 90% of medium Cl- with isethionate. The accompanying fall in input resistance (RN) is therefore probably caused by an increase in K conductance (in agreement with previous reports). There was evidence that anomalous rectification enhances the fall in RN but limits the hyperpolarization. 4. These effects of anoxia were not fully blocked by any of the K-channel antagonists tested, including Cs, TEA, 4-AP, quinine and apamin. 5. Intracellular injections of Ca chelators caused a variable depression of N2-evoked reductions in RN. 6. It is unlikely that N2 activates ATP-sensitive K channels as tolbutamide enhanced rather than depressed the hyperpolarization and fall in RN. 7. When early depletion of cellular ATP was prevented by incubation in creatine (25 mM for greater than 1 h), even longer anoxic periods produced only minor changes in potential, RN, and synaptic transmission. 8. It was concluded that activation of K conductance by a rise in cytosolic-free Ca2+ is the most plausible of several possible underlying mechanisms. PMID- 2547033 TI - Oncogene-related growth factors and growth factor receptors in human malignant glioma-derived cell lines. AB - Oncogenes induce malignant transformation of cells. Two oncogenes are closely related to genes coding for a mitogenic growth factor (v-sis to the PDGF gene) and a receptor for a mitogenic growth factor (v-erb B to the EGF receptor gene). We studied the possibility that cells derived from malignant gliomas produce mitogenic factors that bind to cell surface receptors, the activation of which could lead to excessive stimulation of cell proliferation. All six cell lines tested secrete into their medium factors that stimulate DNA synthesis. The factor secreted by one cell line was characterized and found to resemble PDGF. Six of 11 cell lines had receptors for PDGF demonstrable by binding and receptor autophosphorylation assays. Six of six cell lines tested had EGF receptors demonstrable by binding and receptor autophosphorylation experiments. The extremely high levels of EGF receptor in one cell line may reflect excessive expression of the erb B oncogene associated with abnormalities of chromosome 7 that occur in this cell line. PMID- 2547036 TI - More on [99mTc](V)DMSA scintigraphy in patients with medullary carcinoma of the thyroid. PMID- 2547035 TI - Differential effects of morphine and clonidine on visceral and cutaneous spinal nociceptive transmission in the rat. AB - 1. The effect of morphine or clonidine administered systemically on visceral and cutaneous spinal nociceptive transmission was examined in 45 dorsal horn neurons in spinalized, decerebrate rats: 17 "cutaneous" dorsal horn neurons located in the L3-L5 spinal segments were excited by heating the glabrous skin of the hindpaw (48 degrees C, 15 s) and 28 "visceral" dorsal horn neurons located in the T13-L2 spinal segments were excited by colorectal distension (80 mmHg, 20 s). The 28 visceral dorsal horn neurons were subclassified as 18 short-latency abrupt neurons (SL-A), which were excited by colorectal distension at short latency (less than 1 s) and whose activity abruptly returned to base line following termination of the distending stimulus, and as 10 short-latency-sustained (SL-S) neurons, which also were excited at short latency (less than 1 s) by colorectal distension, but whose activity was sustained above base line for 4-31 s following termination of the distending stimulus. 2. Morphine produced a dose-dependent, naloxone-reversible inhibition of both spontaneous activity and/or neuronal responses during heating or colorectal distension of 8 SL-A, 7 SL-S, and 11 cutaneous dorsal horn neurons. Comparison of the effective doses of morphine to produce a 50% reduction in the response of the neurons (ED50s) during colorectal distension or heating demonstrated that, at the intensities of distension and heating employed, SL-S neurons were affected at the least dosage (ED50 = 0.46 mumol/kg), followed by SL-A neurons (ED50 = 1.95 mumol/kg) and cutaneous neurons (ED50 = 6.12 mumol/kg). Effects on spontaneous activity were variable: at low doses morphine produced an increase in the spontaneous activity of 2 SL-A and 5 cutaneous neurons; greater doses (up to 42 mumol/kg) inhibited in all of the SL-A and SL-S neurons, but not three cutaneous neurons studied. With the exclusion of these three neurons, the ED50s for inhibition of spontaneous activity were comparable to the ED50s for inhibition of neuronal responses during colorectal distension or heating of the hindpaw in all three neuronal groups. 3. Clonidine produced a dose-dependent, yohimbine- or phentolamine-reversible inhibition of both spontaneous activity and neuronal responses during heating or colorectal distension of 10 SL-A, 3 SL-S, and 6 cutaneous dorsal horn neurons.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2547037 TI - Effects of dietary cellulose and psyllium husk on monkey colonic microbial metabolism in continuous culture. AB - The effects of inoculating an in vitro continuous culture system with primate colon contents compared to fecal material, and the effect of feeding these cultures psyllium husk, a fermentable, or cellulose, a less fermentable, dietary fiber were tested. Modified 500-ml Bellco culture chambers were continuously infused with buffered medium containing vitamin mix, deoxycholate, urea, hemin, casein and mucin. Cultures were fed a mixture of minerals, sucrose, starch and either psyllium husk or cellulose twice daily. Chambers were inoculated with fecal or colonic samples obtained from adult male African green monkeys fed the respective fiber source in a purified diet for more than 3 yr. After a 5-d stabilization period, samples were collected for total viable anaerobe and aerobe counts, microbial beta-glucuronidase (EC 3.2.1.31) activity, volatile fatty acid (VFA) and ammonia nitrogen concentrations, dry matter, pH and oxidation-reduction potential. Inoculation with fecal material or colon contents produced similar results for the above mentioned characteristics; major differences were found due to the fiber treatments. Psyllium-fed cultures had lower pH (P less than 0.01) and higher VFA concentration (P less than 0.01) and beta-glucuronidase activity (P less than 0.10) than cellulose-fed cultures. The ratio of anaerobes to aerobes was lower (P less than 0.01) in psyllium-fed than in cellulose-fed cultures. These results indicate that feces can be used as an inoculum source for in vitro studies of changes in colonic microbial metabolism due to diet, and that dietary fiber source affects the colonic microbial population and metabolism. PMID- 2547038 TI - Effects of dietary cellulose, psyllium husk and cholesterol level on fecal and colonic microbial metabolism in monkeys. AB - The effect of long-term feeding of dietary fiber and two levels of cholesterol on monkey colonic microbial metabolism was studied. Three groups of African green monkeys were fed for 3.5 yr purified diets containing 9.7% cellulose or psyllium husk and 0.8 mg cholesterol per kcal or 9.7% cellulose and 0.1 mg cholesterol per kcal. Total viable anaerobe and aerobe counts, microbial beta-glucuronidase activity, volatile fatty acid and ammonia nitrogen concentrations, dry matter and pH were determined in fecal and colonic samples. Compared to cellulose, psyllium husk feeding decreased (P less than 0.05) percentage dry matter, beta glucuronidase (EC 3.2.1.31) activity and pH, and increased (P less than 0.05) ammonia nitrogen and volatile fatty acid output in feces and in colon contents. In all groups, colonic beta-glucuronidase activity was greater (P less than 0.05) than in fecal samples. Microbial beta-glucuronidase activity, pH or percentage dry matter in the ascending colon was not different from that in the transcending or descending segments. The ratio of anaerobic to aerobic bacteria was lower in colon contents from monkeys fed psyllium husk compared to those fed cellulose. Total viable bacterial counts were lower in monkeys fed low cholesterol compared to high cholesterol diets. The results suggest that chronic intake of dietary psyllium husk resulted in greater colonic microbial metabolism compared to cellulose feeding. PMID- 2547039 TI - Haemodynamic effects of acute and chronic renin inhibition in marmosets. AB - We have developed marmoset models for the in vivo evaluation of primate-specific inhibitors of human renin. After acute intravenous administration to normotensive sodium-depleted marmosets, renin inhibitors of different structural types induced a maximum hypotensive response of a magnitude similar to that induced after angiotensin converting enzyme (ACE) inhibition. The response was prevented by pretreatment with an ACE inhibitor. A close relationship between the inhibition of plasma renin activity (PRA) and the fall in blood pressure was observed with most of the inhibitors. CGP 29,287, a synthetic renin inhibitor, and R-3-36-16, a monoclonal antibody, both induced a selective increase in renal blood flow similar to that induced by an ACE inhibitor. A sustained reduction in blood pressure was observed during continuous administration of CGP 29,287 or R-3-36-16 over 14 days, despite an increase in immunoreactive renin and an apparent recovery of PRA. A similar blood pressure fall and an increase in plasma renin was observed during continuous administration of an ACE inhibitor. The renin inhibitor CGP 29,287 also lowered blood pressure after acute administration to hypertensive marmosets with normal PRA. Our studies demonstrate that renin inhibitors have similar haemodynamic effects to ACE inhibitors, and indicate that they may have a similar antihypertensive efficacy. PMID- 2547040 TI - [Evaluation of proliferating activity of pleomorphic adenoma arising from the salivary glands--using anti BrdU monoclonal antibody]. AB - Salivary gland tumors present diverse histopathological aspects and show various biological behavior. At present, the immunohistological method using anti-BrdU monoclonal antibodies, which was originated in the field neurosurgery, has been tried extensively in various fields of medicine. Using this method, the authors have attempted to detect biological characteristics of salivary gland tumor tissues, especially those of pleomorphic adenomas. The labelling index (L.I.) is defined as the percentage of the labelled cells (proliferating cells) in the tumor tissue. In order to confirm the usefulness of the L.I. for measuring the proliferating activity of tumor tissue, an investigation was conducted with respect to squamous cell carcinomas of the head and neck, excluding salivary gland tumors. The results indicate that the L.I. is highly correlated with the differentiation of these carcinomas. In salivary gland tumors, values of the L.I. are also higher for benign tumors than for normal or inflammatory tissues and the values were even higher for malignant tumors. Values of the L.I. are low for pleomorphic adenomas, which are the majority of salivary gland tumors, as well as for other benign tumors. However considerable individual variabilities, are bound for pleom. ad. and the values of the L.I. are markedly high in some cases. These high values are particularly common in patients with whom strong positivity in Ga scintigraphy, or primary growth in the submandibular glands are found. The localization of proliferating cells shows a characteristic pattern in each type of tumor tissue. In pleomorphic adenomas, proliferating cells are sporadically distributed but the main growth is located at the site of the epithelioid proliferative region, excluding the duct-forming locus. However, the overall proliferating activity of myxochondromatous tumors is not generally low, and the proliferating activity is not particularly high in the region of infiltrating is the capsule. PMID- 2547041 TI - IgM to human cytomegalovirus: comparison of two enzyme immunoassays and IgM reactivity to viral polypeptides detected by immunoblotting. AB - IgM detection by enzyme-linked immunosorbent assay (ELISA) is the most used method of establishing a cytomegalovirus (CMV) infection during pregnancy. In this paper, we discuss the results obtained assaying 1,000 sera from pregnant women by two ELISA kits: a traditional indirect ELISA and a more recent IgM capture ELISA. All the sera that gave a positive ELISA value with one or both kits were further tested by immunoblotting (IB) to establish which CMV polypeptides were detected by IgM antibodies. From the results obtained, IgM capture ELISA seems less sensitive than indirect ELISA, but correlates better with serological evidence of active CMV infections as judged by typical IB profiles. PMID- 2547042 TI - Functional and metabolic heterogeneity among normal neutrophil subpopulations. AB - In this study, normal human peripheral blood neutrophils were separated into subpopulations by centrifugation on discontinuous Percoll density gradients and further characterized according to various functional and metabolic capabilities. Chemotaxis was measured in response to optimal concentrations (20 nM) of the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP). According to a population ratio analysis, the majority of cells that contain the highest proportion of fast migrating cells are isolated between the densities of 1.093 and 1.096 gm/ml. Arachidonic acid (AA) metabolism was also determined in neutrophil subpopulations. On initial density gradients, cells isolated from 1.084-1.087 gm/ml and incubated in the presence of exogenous [3H]AA produce 50% of the total [3H]LTB4 produced in response to ionophore A23187 stimulation; whereas after prelabeling with [3H]AA for 2 hr at 37 degrees C, the cells between 1.087 and 1.093 gm/ml are the most active at both releasing [3H]AA from the cellular phospholipids and synthesizing [3H]LTB4 (dpm/cell). These studies demonstrate the significant functional and metabolic heterogeneity that exists among normal neutrophil subpopulations. PMID- 2547043 TI - Primary liver neoplasms: evaluation of proliferative index using MoAb Ki67. AB - Thirty-two cases of primary liver neoplasms comprising 12 benign, 15 malignant, and 5 cases with equivocal histopathological features between benign and malignant have been investigated using monoclonal antibody (MoAb) Ki67, which reacts with a nuclear protein expressed in the G1, G2, S, and M phases of the cell cycle. The Ki67 score (positive cells/total neoplastic cells) seems to correlate to the classes of lesions tested. In the hepatocellular carcinoma (HCC) group, the percentage of labelled nuclei, ranging from 15 to 50 per cent, showed a good correlation with Edmondson-Steiner's histological tumour grade. A percentage of positive cells similar to that of the proved low-grade HCCs was detected in the five neoplastic lesions in which the routine histopathological criteria of malignancy were not fulfilled. The benign neoplasms showed a very low growth fraction, similar to that of normal or cirrhotic tissues. The use of the Ki67 score seems to offer useful information about the biological behaviour of some liver masses and may help in the differential diagnosis of hepatocellular adenoma versus carcinoma. PMID- 2547044 TI - The microsomal glucose-6-phosphatase enzyme of human gall-bladder. AB - Microsomes isolated from adult human gall-bladders have for the first time been shown to contain specific glucose-6-phosphatase activity. The gall-bladder glucose-6-phosphatase enzyme has the same molecular weight (36,500 daltons) and similar immunological properties and kinetic characteristics to the hepatic microsomal glucose-6-phosphatase enzyme. PMID- 2547045 TI - In situ evaluation of the stimulatory state of hepatic macrophages based on their ability to produce superoxide anions in rats. AB - When a liver perfusion with nitro blue tetrazolium (NBT) and phorbol myristate acetate (PMA) was performed in carbon tetrachloride (CCl4)-intoxicated rats, formazan deposition was remarkable in macrophages in the necrotic areas of the liver, its intensity varying with the extent of injury. The deposits almost disappeared after addition of Cu(Lys)2, a scavenger of intra- and extracellular superoxide, but were not affected by superoxide dismutase (SOD), which acts extracellularly. The formazan content after incubation with NBT and PMA was higher in macrophages isolated from CCl4-intoxicated liver than in those from normal liver, though their PMA-induced chemiluminescence did not differ. In Corynebacterium parvum-treated liver, both Cu(Lys)2 and SOD reduced the deposits. This method can estimate in situ the ability of hepatic macrophages to produce superoxide and the cellular sites of its production. PMID- 2547046 TI - Silica-induced pulmonary fibrosis involves the reaction of particles with interstitial rather than alveolar macrophages. AB - Macrophage-derived products have been implicated in fibroblast stimulation following particle deposition in the lung. To assess the role of macrophages in the alveolus versus those in the interstitium in the induction of pulmonary fibrosis, we compared the pulmonary response to silica when phagocytosis occurred predominantly in each of these compartments. One group of mice received intratracheal silica which was phagocytosed largely by alveolar macrophages (AM). A second group was exposed to whole body irradiation prior to receiving the same dose of silica. This prevented the usual efflux of PMN and monocytes into the air sacs, allowing passage of silica particles across the alveolar epithelium to reach the interstitial macrophages (IM). In the irradiation plus silica group, many large interstitial granulomas were formed at 2-4 weeks, and collagen levels were significantly greater than in all other groups at 16 weeks. More silica was found in a lung tissue residue and in lymph nodes of these animals. Pulmonary fibrosis was limited to interstitial areas where there was a high level of retained silica, whereas peripheral regions of the lung, where free AM containing silica were found, did not show fibrosis of the alveolar walls. The results suggest that factors secreted by IM in response to silica are more effective in stimulating fibrogenesis than secretions made by the AM into the alveolar space. PMID- 2547047 TI - T-cell antigen receptor beta-chain variable region families: a study of their distribution in normal and reactive tissues. AB - The beta chain of the T-cell antigen receptor is constructed using a variable region gene from one of approximately 20 variable gene families. Antibodies that react with the products of these gene families have been used to demonstrate clonal proliferation of T cells. Here, we describe the expression of two beta chain variable region families in normal and reactive lymphoid tissues. In all the tissues studied, expression was scattered throughout the T-cell areas, without any clustering of expression. PMID- 2547048 TI - Vimentin--a new prognostic parameter in breast carcinoma? AB - This report describes a positive relationship between vimentin expression in infiltrating ductal breast carcinoma, and high tumour growth fraction. Vimentin expression is potentially a predictor of aggressive behaviour, and such carcinomas may benefit from early adjuvant therapy. Eighty-four malignant breast neoplasms were stained with monoclonal anti-vimentin and anti-cytokeratin antibodies. The tumour growth fractions were determined by immunostaining cryostat sections with the Ki-67 antibody. Seven (9.2 per cent) of 76 infiltrating ductal carcinomas co-expressed cytokeratin and vimentin intermediate filaments in more than 50 per cent of neoplastic cells. In each case, the corresponding Ki-67 count was much greater than 40 per cent, significantly higher than the mean growth fraction for all tumours examined (P less than 0.0001). Vimentin immunoreactivity was also positively related to the histological grade of the ductal carcinomas (P less than 0.002) and inversely related to tumour ER count (P less than 0.0002) and patient age (P less than 0.01). No relationship was observed between vimentin positivity and either the presence of axillary nodal metastases or primary tumour size. PMID- 2547049 TI - Undifferentiated columnar cells in colorectal adenomas and familial adenomatous polyposis. AB - We have observed and defined morphometrically and histochemically groups of undifferentiated columnar cells within the surface epithelium in colorectal mucosa. They were present within both non-polypoid and polypoid mucosa in familial adenomatous polyposis, and within non-hereditary adenomatous polyps of the colon and rectum. The cells show some evidence of proliferative activity and appear similar to cells previously described in the stomach which were proposed as precursors to type 3 sulphomucin-secreting intestinal metaplasia in atrophic gastritis. To our knowledge, these observations have not been previously described. It is possible that the cells represent the cellular basis of the shift in the proliferative zone from the normal site at the crypt base to the colorectal mucosal surface, which is known to precede adenomatous polyp formation. The cells may therefore be involved in the early stages of colorectal adenoma formation. PMID- 2547050 TI - Human herpesvirus type 6 infection (exanthem subitum) without fever. PMID- 2547051 TI - Selective IgA deficiency temporally associated with Epstein-Barr virus infection. PMID- 2547052 TI - Inflammatory cell function in young rodents with experimental cholestasis: investigations of functional deficits, their etiology, and their reversibility. AB - Children with cholestasis are susceptible to infective complications. This may be attributable to impaired host defense. We postulated that cholestasis affects systemic polymorphonuclear leukocyte (PMN) function by impeding chemotaxis, phagocytosis, and superoxide release, which are all critical in eliciting an adequate immune response. Sprague Dawley rats (225 g) were assigned to three groups: bile duct ligated (BDL), sham (SH), and normal control (NC). On day 21 after operation, PMN and sera were isolated. Chemotactic response to C5a and FMLP (formyl-methionyl-leucyl-phenylalanine), superoxide release, and phagocytic uptake of 14C-labeled Staphylococcus aureus were performed on pooled PMN samples. Results were expressed as mean +/- SD. Serum bilirubin at day 21 was 6.3 +/- 2.9 v 0.1 +/- 0.1 and 0.1 +/- 0 mg/dL (P less than .01) in BDL, SH, and NC groups, respectively. Kinetic studies of PMN phagocytosis demonstrated impaired 14C S aureus uptake by BDL neutrophils at 60 (P less than .05), 90 (P less than .05), and 120 minutes (P less than .05) compared with SH and NC groups. No differences in PMN chemotactic response to C5a and FMLP was observed in BDL, SH and NC groups (43 +/- 14 v 40 +/- 12 and 33 +/- 1, and 43 +/- 20 v 43 +/- 14 and 28 +/- 1 cell per field, respectively). Zymosan stimulated superoxide release did not differ between groups (14.3 +/- 3.6 (BDL) v 15.1 +/- 8.7 (SH) and 12 +/- 2.0 (NC) nmol/30 min/mg cell protein, respectively. Thus, cholestasis impairs neutrophil phagocytosis in vitro.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547053 TI - Two decades of experience with testicular tumors in children at St Jude Children's Research Hospital. AB - From 1968 to 1988, 24 children and adolescents with malignant testicular tumors were treated at St Jude Children's Research Hospital. Pure yolk sac tumors (YST) were present in 13 cases; 11 patients had other types of nonseminomatous malignant germ cell tumors. Children with localized and totally resectable disease (stage I) were treated by orchiectomy alone; all others also received chemotherapy. Five of ten patients treated before the implementation of a multiagent chemotherapy protocol in 1979 have died. By contrast, all of the 14 patients treated on this protocol are alive. The improved survival during the past decade is attributable to better diagnostic imaging techniques, the availability of serum tumor markers to monitor disease activity, and more effective chemotherapy. Orchiectomy alone is sufficient treatment for patients with clinical stage I disease who show appropriate reductions in tumor marker levels after surgery. Modern platinum-based chemotherapy provides disease control in patients with higher stage disease. PMID- 2547054 TI - Use of intraoperative ultrasound during hepatic resection in pediatric patients. AB - Five pediatric patients with primary liver tumors were evaluated preoperatively with ultrasound (US), computerized tomography (CT), and angiography, and tentative operative plans were formulated. Intraoperative US was subsequently used to examine these children, resulting in changes in operative strategy of all five patients despite their extensive preoperative evaluations. Intraoperative ultrasound appears to provide the most accurate assessment of both the extent of tumor and its vascular relationships. Thus, operative strategies may be precisely tailored on the basis of such information, allowing rational resection where appropriate, while futile attempts at removal of inoperable lesions may be averted. PMID- 2547055 TI - Measles outbreaks: who are at risk and why. AB - After the introduction of measles vaccine in the United States in 1963 the reported incidence of measles (rubeola) decreased substantially. The disease, however, has not been eliminated. Since 1983, when the lowest number of cases was reported, slight increases in incidence have been observed. Outbreaks are occurring among previously immunized school- and college-age children and unimmunized preschool children and infants. This article describes measles occurrence, transmission, diagnosis, development of measles immunity, the 1989 Centers for Disease Control recommendations for immunization, and implications for health care providers for preventing measles outbreaks. PMID- 2547057 TI - Continuous naloxone administration suppresses opiate withdrawal symptoms in human opiate addicts during detoxification treatment. AB - In a small clinical trial, a new therapeutic approach was studied, whether naloxone, in high dosage over a prolonged period of time, will attenuate withdrawal symptoms in acute opiate detoxification. Six opiate addicts, satisfying DSM III-R criteria of opiate dependence, were given 10 mg naloxone under short barbiturate anaesthesia, followed by repeated doses of 0.4 mg/h naloxone for at least 72 h. Acute onset of withdrawal symptoms brought about a high dose of naloxone could be suppressed by the short barbiturate anaesthetic; neither continuous supply nor cessation of the naloxone regimen after 96 h caused any severe withdrawal symptoms. Morphine and naloxone measurements in blood at the start of naloxone therapy enabled pharmacokinetic explanations for this paradoxical action of naloxone to be excluded. PMID- 2547056 TI - Experimental infection of cynomolgus monkeys (Macaca fascicularis) with simian immunodeficiency virus (SIVsm). AB - Five healthy cynomolgus monkeys were inoculated intravenously with simian immunodeficiency virus (SIVsm) propagated in human lymphocytes. All five animals became infected. Virus was recovered from blood mononuclear cells and viral antigen was detected in serum 12 days postinoculation (PI) in all inoculated animals. Virus was also isolated in all five animals tested 74 to 226 days PI. Antibodies to different structural proteins of SIV and HIV-2 were demonstrated by ELISA, Western blot, and radioimmunoprecipitation assay from day 31 PI concomitantly with a reduction of viral proteins in the serum. Reappearance of antigen accompanied by a fall in antibody to gag products (p26) was observed in two monkeys 69 days PI. All SIV-infected monkeys showed a pronounced decrease in CD4+ lymphocytes demonstrable already 12 days PI. They also developed persistent lymphadenopathy. Thus, infection of cynomolgus monkeys with SIVsm mimics events in human immunodeficiency virus infection in humans but the course of evolution of pathogenic events in the monkey is markedly compressed. This experimental model will be useful for evaluation of HIV vaccines and antiviral testing. PMID- 2547058 TI - Flagellar movement in demembranated preparations of ejaculated fowl spermatozoa. AB - Triton X-100 at a concentration of 0.1% in the extraction medium was optimal for demembranating fowl spermatozoa. The most suitable conditions for reactivation were obtained when a medium composed of 0.5 mM-ATP, 25 mM-potassium glutamate, 10(-7) M-CaCl2, 20 mM-Tris-HCl(pH 7.9), 1 mM-MgSO4, 1 mM-dithiothreitol and 0.2 M sucrose was used. More than 60% motile spermatozoa were obtained under these conditions. The addition of 1 or 10 microM-cAMP did not appreciably affect motility. Intact and demembranated spermatozoa were immotile at 40 degrees C, whilst at 30 degrees C motility was restored. PMID- 2547059 TI - Influence of A23187 and dibutyryl cyclic AMP on progestagen production by rat granulosa cells in vitro. AB - The gonadotrophic regulation of progesterone production by rat granulosa cells was examined in a chemically-defined medium containing FSH, dibutyryl cyclic AMP [Bu)2cAMP) and the calcium ionophore, A23187. FSH and A23187 alone significantly enhanced the production of pregnenolone, progesterone and its metabolite, 20 alpha-hydroxypregn-4-en-3-one (20 alpha-OH-P) from endogenous substrate(s). Stimulation of progesterone production by A23187 was accompanied by an increase in 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) but not 20 alpha hydroxysteroid dehydrogenase (20 alpha-HSD) activity, as attested by enhancement of the metabolism of exogenous pregnenolone to progesterone but not of progesterone to 20 alpha-OH-P. In contrast, although (Bu)2cAMP increased pregnenolone and progesterone production and the metabolism of exogenous progesterone to 20 alpha-OH-P, it failed to stimulate the conversion of exogenous pregnenolone to progesterone. The increase in progesterone production and in the conversion of exogenous pregnenolone to progesterone by FSH and A23187 was concentration- and time-dependent. Whereas maximal stimulation of de-novo progesterone synthesis by FSH was evident by 6 h (earliest time examined), a significant increase in the conversion of exogenous pregnenolone to progesterone in the presence of FSH or the ionophore was not noted until 12 h of incubation. Although a small but significant increase in progesterone production was also noted as early as 6 h of incubation in the presence of the calcium ionophore, this was markedly smaller than that elicited by FSH. We conclude that the calcium ionophore A23187 and (Bu)2cAMP have similar as well as distinct effects on progesterone production in rat granulosa cells in vitro.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547061 TI - Systemic lupus erythematosus and concurrent cytomegalovirus vasculitis: diagnosis by antemortem skin biopsy. AB - A 45-year-old woman with a 4-year history of systemic lupus erythematosus (SLE) developed fever, decreased visual acuity and skin ulceration. A biopsy of a cutaneous ulcer demonstrated small vessel vasculitis with characteristic cytomegalovirus (CMV) inclusions in the vascular endothelium. The presence of CMV was confirmed by DNA hybridization immuno-histochemistry. Retinal artery vasculitis, previously associated with flares of her SLE, was also noted on ophthalmologic examination. Our case demonstrates that CMV infection can mimic the cutaneous manifestations of collagen vascular disease and that early identification can be made by biopsy of suspicious skin lesions. PMID- 2547060 TI - Antiperinuclear factor, a rheumatoid arthritis specific autoantibody: its relation to Epstein-Barr virus. AB - We studied the prevalence of antiperinuclear factor (APF), an antibody frequently (50-80%) present in sera of patients with rheumatoid arthritis (RA), in different groups of patients. Sera from 123 patients with RA and 28 patients with recent Epstein-Barr virus (EBV) infection showed APF in 63 and 51% of the patients, respectively. These frequencies were significantly increased (p0.001) when compared with 123 healthy blood donors (12%) and in 58 patients with herpes virus infections other than EBV (18%). Relations with other EBV elicited antibodies in RA and the relevance of EBV in RA are discussed. Our data suggest that EBV might be the immunogen for APF, an antibody, until now considered to be an autoantibody. PMID- 2547062 TI - An association between human parvovirus B-19 infection and autoantibody production. PMID- 2547063 TI - AIDS: neurological opportunist infections in central London. AB - Twenty six (41%) of 64 central London cases of AIDS with nervous system involvement during the course of the illness had neurological opportunist infection. Cytomegalovirus and Toxoplasma gondii were the commonest agents in 22 cases with central nervous system (CNS) infection. Eight cases had herpes zoster radiculopathy. Other infections included those caused by Cryptococcus neoformans, Mycobacterium tuberculosis and papova JC virus. Prognosis was generally poor, irrespective of whether the opportunist infection was treatable. PMID- 2547064 TI - Antibody imaging to locate a placental site trophoblastic tumour following a complete hydatidiform mole. PMID- 2547065 TI - Retroviruses and diseases of the nervous system. PMID- 2547066 TI - Bovine visna virus and the origin of HIV. PMID- 2547067 TI - Localization of insulinoma by intraoperative ultrasonography. PMID- 2547068 TI - Replacement of the 3'-CH group by nitrogen in the carbocyclic analogue of thymidine. AB - We have prepared (4R)-4-thyminyl-D-prolinol, an analogue of 3'-deoxythymidine in which the sugar has been replaced by D-prolinol. This strongly basic secondary amine has been converted to the corresponding hydroxylamine, an analogue of either thymidine or 2'-deoxyxylofuranosylthymine. We have also synthesized a number of simple derivatives of the amine for testing in vitro activity against herpes simplex 1 (HSV-1), human immunodeficiency virus 1 (HIV-1), and a panel of human tumor cell lines. Among these compounds, the hydroxylamine 12 proved active against the human tumor cell lines of breast, colon, and lung origin, with IC50 values of 0.08, 14.02, and 6.91 microM, respectively. PMID- 2547069 TI - Synthesis and antirhinovirus activity of 6-(dimethylamino)-2-(trifluoromethyl)-9 (substituted benzyl)-9H-purines. AB - A series of 6-(dimethylamino)-2-(trifluoromethyl)-9-(substituted benzyl)purines was synthesized and tested for antirhinovirus activity. Most of the compounds were synthesized by alkylation of 6-chloro-2-(trifluoromethyl)-9H-purine with the appropriate benzyl halide followed by displacement of the chloro group with dimethylamine. Alternatively, 6-(dimethylamino)-2-(trifluoromethyl)purine was alkylated with the appropriate benzyl halide. Although several different aryl substituents provided compounds with IC50's = 0.03 microM against rhinovirus serotype 1B, no congener was significantly more active than the parent 2. Twenty three compounds were tested against 18 other serotypes, but none exhibited a uniform profile of activity. PMID- 2547070 TI - Synthesis and benzodiazepine receptor affinities of rigid analogues of 3-carboxy beta-carbolines: demonstration that the benzodiazepine receptor recognizes preferentially the s-cis conformation of the 3-carboxy group. AB - 1H-Indolo[3',2':4,5]pyrido[3,2-b]-2-penten-5-olide (6) and 1H,5H-indolo[3',2'-c] 6,7-dihydro-2-pyridone (7), rigid analogues of methyl 4-ethyl-beta-carboline-3 carboxylate (8) and N-methyl-4-ethyl-beta-carboline-3-carboxamide (9), respectively, were synthesized and their in vitro binding affinities to the central type benzodiazepine receptors were compared. The IC50 values of 6 and 8 were approximately equivalent (42 and 27 nM, respectively). The amide derivative 9, for which theoretical energy calculations indicate that the s-trans carbonyl conformation is the preferred one, displayed very low affinity (IC50 greater than 10(4) nM). However, when the carbonyl group of 9 was forced to adopt the s-cis conformation as in lactam 7, binding to the benzodiazepine receptor was largely restored (IC50 = 150 nM), indicating that the s-cis carboxy conformation at C-3 of beta-carbolines is preferentially recognized by this receptor. In vivo, compound 6 showed neither convulsant, proconvulsant, nor anticonvulsant activity in mice. Moreover, 6 did not antagonize methyl beta-carboline-3-carboxylate induced convulsions in mice. This lack of activity of 6 was attributed to its inability to cross the blood-brain barrier since no significant displacement of [3H]Ro 15-1788 from mouse brain benzodiazepine receptors by 6 could be observed in vivo. PMID- 2547071 TI - 3,4-Dihydro-2H-1-benzopyran-2-carboxylic acids and related compounds as leukotriene antagonists. AB - Evaluation of a series of 3,4-dihydro-2H-1-benzopyran-2-carboxylic acids linked to the 2-hydroxyacetophenone pharmacophore present in the standard peptidoleukotriene antogonist FPL 55712 (1) has led to the discovery of Ro 23 3544 (7), an antagonist possessing greater potency and duration of action vs LTD4 than the standard (aerosol route of administration, guinea pig bronchoconstriction model). Interestingly, this compound also potently inhibited bronchoconstriction induced by LTB4 whereas 1 did not. Attempts to establish structure--activity relationships in this series involved modifications in the 2 hydroxyacetophenone moiety, the linking chain, and the chroman system. All variations produced analogues which were either inactive or possessed reduced potency relative to acid 7. Optical resolution of 7 was achieved by two methods. Absolute configurations of the enantiomers were determined via X-ray crystallographic analyses of an intermediate as well as a salt of the S enantiomer. Although the enantiomers exhibited similar potencies in in vitro assays and in vivo when administered intravenously, significant differences were observed in the guinea pig bronchoconstriction model vs LTC4 and LTD4 when administered by the aerosol route (S antipode 15-fold more potent). The properties of 7 have been compared with several recently reported leukotriene antagonists. PMID- 2547072 TI - Synthesis and antiviral activity of the enantiomeric forms of carba-5-iodo-2' deoxyuridine and carba-(E)-5-(2-bromovinyl)-2'-deoxyuridine. AB - Both enantiomers of the carbocyclic analogues of 5-iodo-2'-deoxyuridine (14 and ent-14) and of (E)-5-(2-bromo-vinyl)-2'-deoxyuridine (16 and ent-16) were synthesized by using (+)- or (-)-endo-norborn-5-en-2-yl acetate or butyrate, respectively, as starting materials. Against herpes simplex virus type 1 (+)-C BVDU (16) was only slightly less active than BVDU itself, whereas (-)-C-BVDU (ent 16) proved to be 10-400-fold less effective, depending on the strain investigated. Against HSV-2 both (+)- and (-)-C-BVDU as well as (+)- and (-)-C IDU showed minor activity. All carbocyclic analogues were inactive against TK-HSV 1 strains, pointing to the prerequisite of phosphorylation (activation) by the viral thymidine kinase (TK). PMID- 2547073 TI - 2-[[(4-Amino-2-pyridyl)methyl]sulfinyl]benzimidazole H+/K+-ATPase inhibitors. The relationship between pyridine basicity, stability, and activity. AB - The benzimidazole sulfoxide class of antisecretory H+/K+-ATPase inhibitors need to possess high stability under neutral physiological conditions yet rearrange rapidly at low pH to the active sulfenamide 2. Since the initial reaction involves internal nucleophilic attack by the pyridine nitrogen, control of the pyridine pKa is critical. In this paper we show that by utilizing the powerful electron-donating effect of a 4-amino substituent on the pyridine, moderated by the electron-withdrawing effect of a 3- or 5-halogen substituent, a combination of high potency (as inhibitors of histamine-stimulated gastric acid secretion) and good stability under physiological conditions can be obtained. Furthermore, the role of the steric interaction between the 3/5-substituents and the 4 substituent in modifying the electron-donating ability of the 4-amino group is exemplified, and additional factors affecting stability are identified. One compound, in particular, 2-[[(3-chloro-4-morpholino-2- pyridyl)methyl]sulfinyl]-5 methoxy-(1H)-benzimidazole (3a, SK&F 95601), was chosen for further development and evaluation in man. PMID- 2547074 TI - Alterations in the stereochemistry of the kappa-selective opioid agonist U50,488 result in high-affinity sigma ligands. AB - The synthesis and in vitro sigma receptor activity of the two diastereomers of U50,488 [(+/-)-2], namely, (1R,2S)-(+)- cis-3,4-dichloro-N-methyl-N-[2-(1 pyrrolidinyl)cyclohexyl]benzeneacet ami de [(+)-1] and (1S,2R)-(-)-cis-3,4 dichloro- N-methyl-N-[2-(1-pyrrolidinyl)cyclohexyl]benzeneacetamide [(-)-1], are described. (+)-1 and (-)-1 were synthesized from (+/-)-trans-N-methyl-2 aminocyclohexanol [(+/-)-3]. Pyridinium chlorochromate (PCC) oxidation of the N-t Boc-protected derivative of (+/-)-3 afforded (+/-)-2-[N- [(tert butyloxy)carbonyl]-N-methylamino]cyclohexanone [(+/-)-5]. The sequence of enamine formation with pyrrolidine, catalytic reduction, N-deprotection, and optical resolution afforded (1R,2S)-(-)-cis-2-pyrrolidinyl-N-methylcyclohexylamine [(-) 10] and (1S,2R)-(+)-cis-2-pyrrolidinyl-N-methylcyclohexylamine [(+)-10]. The optical purity (greater than 99.5%) of (-)-10 and (+)-10 was determined by HPLC analysis of the diastereomeric ureas formed by reaction with optically pure (R) alpha-methylbenzyl isocyanate. The absolute configuration of (-)-10 and (+)-10 was determined by single-crystal X-ray diffractometry of the bis-(R)-mandelate salt. Condensation of optically pure (-)-10 and (+)-10 with 3,4 dichlorophenylacetic acid furnished (+)-1 and (-)-1, respectively. Compounds (+) 1, (-)-1, (-)-2, and (+)-2 were compared for their binding affinities at kappa opioid, sigma, D2-dopamine, and phencyclidine (PCP) receptors in competitive binding assays using [3H]bremazocine ([3H]BREM) or [3H]U69,593, [3H]-(+)-3-(3 hydroxyphenyl)-N-(1-propyl)piperidine [[3H]-(+)-3-PPP], or [3H]-1,3-di(o tolyl)guanidine ([3H]DTG), [3H]-(-)-sulpiride [[3H]-(-)SULP], and [3H]-1- [1-(2 thienyl)cyclohexyl]piperidine ([3H]TCP), respectively. In the systems examined, ( )-2 exhibited the highest affinity for kappa receptors, with a Ki of 44 +/- 8 nM. However, (-)-2 also showed moderate affinity for sigma receptors, with a Ki of 594 +/- 3 nM [[3H]-(+)-3-PPP]. The (1R,2R)-(+)-enantiomer, (+)-2, had low affinity for both kappa and sigma receptors, exhibiting Ki values of 1298 +/- 49 nM at kappa ([3H]BREM) and 1270 +/- 168 nM at sigma [[3H]-(+)-3-PPP]. In contrast, the chiral cis compounds (+)-1 and (-)-1 showed high affinity for sigma receptors and negligible affinity for kappa opioid receptors in the [3H]BREM assay. Compound (-)-1 exhibited a Ki of 81 +/- 13 nM at sigma receptors [[3H]-(+) 3-PPP] and 250 +/- 8 nM ([3H]DTG).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2547075 TI - New model for the secondary structure of the 5' non-coding RNA of poliovirus is supported by biochemical and genetic data that also show that RNA secondary structure is important in neurovirulence. AB - A secondary structure model for the 5' non-coding RNA of poliovirus has been derived by comparing computer-generated folding patterns of equivalent sequences from a number of related enteroviruses and rhinoviruses and identifying compensating mutations that suggest conservation of a common secondary structure. Although certain elements are similar, the new model differs considerably from a previously published minimal energy structure and is consistent with the observed sensitivity of in vitro RNA transcripts of infectious poliovirus cDNA to RNases and modifying chemicals. The sequence of a neurovirulent revertant of an attenuated mutant provides additional evidence for an interaction between a region known to be important for neurovirulence, sequence 471-483, and nucleotides 528 to 538. PMID- 2547076 TI - Identification and description of beta-structure in horse muscle acylphosphatase by nuclear magnetic resonance spectroscopy. AB - Nuclear magnetic resonance spectra of acylphosphatase were searched for signs of beta-structure, i.e. characteristic nuclear Overhauser enhancement patterns displayed in the two-dimensional spectra, typical chemical shifts, coupling constants and slow 2H-H exchange. The results provided identification of the main chain resonances of amino acid residues involved in the beta-structure. The full sequential assignment of this region was gained by identification of some amino acid spin systems and their alignment with the primary sequence. The assignment of the side-chains was virtually completed subsequently and a list produced of nuclear magnetic resonance (n.m.r.) constraints derived from the spectra. The beta-structure consists of a beta-sheet with four antiparallel chains, one attached parallel chain, three tight turns and a beta-bulge. The conformation of the beta-sheet was determined by distance geometry calculation using the n.m.r. constraints (174 intraresidual, 107 sequential and 226 long-range distances, 32 torsion angles, phi, and 28 hydrogen bonds) as input. Observation of some interactions between the sheet and previously identified alpha-helical regions made it possible to give an outline of the three-dimensional structure of the enzyme. PMID- 2547077 TI - In vitro spinal cord conduction block during exposure to a xanthine oxidase/hypoxanthine system: noninvolvement of superoxide and hydrogen peroxide. AB - The effects of a reactive oxygen system on axonal conduction were assessed in an in vitro rat spinal cord preparation. An enzyme system, containing hypoxanthine and xanthine oxidase as a source of superoxide and hydrogen peroxide, was used in combination with ADP and FeCl3 as catalysts for peroxidative activity. The reactants were mixed as they entered a temperature-controlled Plexiglas chamber containing a longitudinal hemisection of adult rat spinal cord. Extracellular action potentials were recorded with a glass microelectrode before, during, and after the exposure. A significant conduction block developed during the 30 min exposure. Action potential amplitude decreased to less than 45% of pre-exposure level while absolute refractory period to paired stimuli increased 160%. Following reintroduction of normal bathing medium, amplitude and absolute refractory period exhibited recovery toward pre-exposure control levels, but did not fully recover. Isolated spinal cord membranes exposed to the same xanthine oxidase system produced significant levels of malondialdehyde (MDA). Superoxide dismutase (SOD), but not catalase, effectively inhibited MDA production. Hypoxanthine, xanthine oxidase, and ADP-Fe3+ were all required to induce conduction block in the spinal cord and peroxidation in the isolated membranes. However, addition of intermediate scavengers, SOD and catalase, alone or in tandem, did not prevent the conduction block. Mechanisms other than radical induced lipid peroxidation may be working to alter the membrane ionic equilibrium in the cord preparation. PMID- 2547078 TI - Dietary habits and prognostic factors in breast cancer. AB - The relationship between dietary habits and prognostic factors for breast cancer was studied in 240 women aged 50-65 years who had surgery for breast cancer between 1983 and 1986. A dietary history interview was conducted within the 4 months following resection of the primary tumor. In the stepwise multivariate analysis, the multiple-odds ratio (OR) for having a tumor greater than or equal to 20 mm in diameter was 0.95 (95% confidence interval, 0.91-0.99) for each 1-g increase in fiber intake per 10 MJ of energy intake. Compared with patients having tumors poor in estrogen receptor (ER), those having ER-rich tumors (greater than or equal to 0.10 fmol/microgram of DNA) were older (P less than .01) and reported carbohydrate intake yielding higher E% (percentage of total energy intake) (P less than .01) and higher retinol intake per 10 MJ (P less than .05). The OR for having an ER-rich tumor was 1.58 (95% confidence interval, 1.08 2.31) for each 1-mg increase in retinol intake per 10 MJ; 1.09 (95% confidence interval, 1.02-1.16) for each additional year of age; and 1.08 (95% confidence interval, 1.02-1.13) for each 1% increment in E% from carbohydrates. These results suggest that the dietary patterns of the western world (e.g., high fat intake and low intake of carbohydrates and fiber) affect certain prognostic factors in breast cancer, such as tumor size and ER content of the tumor. PMID- 2547079 TI - Urinary excretion of cyclic AMP in cadmium-intoxicated rats. AB - To obtain further information on the negative calcium balance caused by Cd, the factors associated with serum calcium and phosphorus homeostasis other than inhibition of intestinal calcium absorption were studied by using urinary cyclic 3',4'-adenosine monophosphate (cAMP). In rats exposed to Cd for 30 d, the levels of urinary excretion of cAMP after treatment with parathyroid hormone (PTH), parathyroidectomy (PTX), or 1 alpha-hydroxycholecalciferol (1 alpha-OH-D3) showed almost the same patterns as those of control rats: the response of urinary cAMP to treatment with PTH was not influenced by continuous oral administration of Cd. On the other hand, in rats exposed to Cd for 90 d without the other three treatments, the amount of urinary excretion of cAMP was markedly higher than in control rats. In PTX rats exposed to 90 d of Cd, urinary cAMP was unchanged, but it was markedly increased when the parathyroid was intact, with or without treatment with PTX. This phenomenon indicated hyperparathyroidemia in response to continuous oral administration of Cd for 90 d. The negative calcium balance with hyperparathyroidemia occurred after continuous oral administration of Cd and developed via increased urinary excretion of calcium. Urinary excretion of cAMP in Cd-exposed rats was unaffected by the administration of 1 alpha-OH-D3. PMID- 2547080 TI - Cadmium-enriched cigarette smoke-induced cytological and biochemical alterations in rat lungs. AB - Male Sprague-Dawley rats were exposed daily for 52 wk in a nose-only exposure system to smoke from the University of Kentucky 2R1 reference cigarettes (SM) or from cigarettes made of cadmium-enriched tobacco (Cd-SM). At sacrifice, the animals were evaluated by bronchoalveolar lavage (BAL) for inflammatory cell response in the lungs, and the cells so obtained were analyzed for phagocytosis of particles (latex and IgG-coated SRBCs) and for their ability to release oxidants upon phagocytic challenge. Additionally, lung tissues were analyzed for Cd levels and lung homogenate fractions were assayed for aryl hydrocarbon hydroxylase (AHH) as well as total and selenium-dependent glutathione peroxidase (GSH-Px) activities. BAL cell counts showed a significant influx of inflammatory cells into the lungs of the Cd-SM group but not the SM group. The proportion of neutrophils in the BAL cells of the Cd-SM group was elevated to 40 +/- 9%, compared with less than 2% in the SM group. Phagocytosis of both types of particles by macrophages from SM and Cd-SM groups was similar to that of the control groups, except that a greater uptake of latex particles was seen in Cd-SM macrophages. The release of oxidants (superoxides and hydrogen peroxide) by the BAL cells was severely impaired in the Cd-SM group, whereas a slight stimulation was seen in the SM group. Pulmonary GSH-Px activity was the same in all groups. A significant induction of the pulmonary AHH activity was observed in the SM group only. The Cd levels in the lungs were approximately 8- and 200-fold greater than controls in SM and Cd-SM groups, respectively. These observations suggest a significant influence of tobacco Cd on the toxicity of cigarette smoke. PMID- 2547081 TI - In vitro and in situ inhibition of the sodium channel blocker saxitoxin by monoclonal antibodies. AB - The sodium channel blocker saxitoxin (STX) was conjugated to keyhole limpet hemocyanin (KLH) and used to immunize BALB/c mice. Anti-STX antibodies were detected in serum by an enzyme-linked immunosorbent assay (ELISA) within a week or two after the first immunization. Spleens from immunized mice were fused with NS-1 myeloma cells and approximately 7000 resultant hybrids were screened by ELISA for reactivity to STX. Two stable hybrids were isolated, subcloned, and characterized. These hybrids, termed S1A5 and S3E.2, secreted specific anti-STX antibodies that did not recognize the closely related toxin tetrodotoxin (TDT), as determined by competition ELISA. The S1A5 monoclonal antibody (mAb) was of the IgMk class and S3E.2 of the IgG1k subclass with affinity constants (Ka values) of approximately 10(6) M-1. The protective ability of these antibodies was tested by a competitive displacement assay for [3H]STX binding on rat brain membranes. Purified S3E.2 strongly displaced [3H]STX binding, whereas S1A5 weakly inhibited [3H]STX binding to membranes. One nanomole of S3E.2 or S1A5 was able to bind 0.03 nmol or 0.005 nmol, respectively, of STX. The S3E.2 mAb offered partial protection against STX-induced reduction of peripheral nerve action potential in rat tibial nerve when administered in situ at concentrations 10- to 30-fold greater than STX. The S1A5 mAb, despite its ability to inhibit STX binding in vitro, was completely ineffectual in situ. These antibodies, particularly S3E.2, thus represent potentially useful reagents for neurobiologic research, detection of toxin contamination, and diagnosis of poisoning, and may provide protection against the toxicity of STX in vivo. PMID- 2547082 TI - On the GABAA receptor: a molecular modeling approach. AB - Bicuculline methobromide, a complex alkaloid, antagonizes in some unknown manner the action of GABA, an important inhibitory transmitter in the CNS. To help understand the mechanism of this antagonism we have employed molecular modeling techniques to assess the similarity and difference between this antagonist and GABA plus four direct GABA agonists. Topological and electronic charge congruence between these five molecules was examined in great detail. It was found that each of the five molecules has three clearly defined atoms that serve as attachment points at the GABAA receptor site. It is hypothesized that an additional negatively charged atom on bicuculline serves as an additional point of attachment that blocks the nearby chloride ion channel. The model presented offers an explanation of why muscimol acts as a better agonist than GABA as well as rationalizing why (+)-bicuculline acts as an antagonist but (-)-bicuculline does not. PMID- 2547083 TI - Spontaneous and beta-adrenergic receptor-mediated taurine release from astroglial cells do not require extracellular calcium. AB - Astroglial cells release taurine when stimulated by beta-adrenergic agonists and other neuroactive agents. The Ca2+-dependency of taurine release by an LRM55 astroglial cell line was investigated by removing Ca2+ from the perfusion medium and by using three inorganic and three organic Ca2+-channel blockers (Mn2+, Co2+, Cd2+, verapamil, nifedipine, and diltiazem). Spontaneous release and release stimulated by the beta-adrenergic agonist isoproterenol were not inhibited when cells were perfused with medium containing no added Ca2+ and 10 microM EGTA. Isoproterenol-stimulated taurine release was not blocked when extracellular Ca2+ was completely replaced by Mn2+, Co2+, or Cd2+, nor was it blocked by verapamil, nifedipine, or diltiazem. In fact isoproterenol-stimulated taurine release was increased by 50 microM diltiazem and when Ca2+ was replaced by Co2+. The rate of spontaneous release increased slowly and continually when Co2+ was substituted for Ca2+ but was almost unaffected by substitution of Mn2+ or Cd2+. Application of diltiazem increased spontaneous release significantly, while verapamil and nifedipine appeared to cause small increases. These results indicate that entry of Ca2+ from the extracellular medium is not required for either receptor mediated or spontaneous taurine release from astroglial cells. Some other changes in the medium did strongly affect release. Both spontaneous and isoproterenol stimulated release were inhibited by elevated osmotic pressure, and spontaneous release was greatly increased when Ca2+ was completely removed without substituting another divalent cation. Spontaneous release increased when antagonistic metal ions were replaced with Ca2+ and when organic channel blockers were removed. PMID- 2547084 TI - Neurotrophic activity of monomeric glucophosphoisomerase was blocked by human immunodeficiency virus (HIV-1) and peptides from HIV-1 envelope glycoprotein. AB - Glucophosphoisomerase (GPI), a glycolytic enzyme, was recently described to share 90% sequence homology with neuroleukin, a recently discovered growth factor which promotes motor neuron regeneration in vivo, survival of peripheral and central neurons in vitro, and affects B cell immunoglobulin synthesis. Interestingly, neuroleukin activity was described to be antagonized by the human immunodeficiency virus (HIV-1) envelope glycoprotein (gp120), with which neuroleukin was found to share partial sequence homology. In this study, reduced GPI demonstrated similar activity to neuroleukin in a novel bioassay using human and rat neuroblastoma cell lines. In the presence of reduced GPI, these cells were found to differentiate, in terms of enhanced neurite extension at a reduced proliferation rate. These results demonstrate the existence of a novel growth factor activity of an evolutionary ancient enzyme. The nonreduced commercial form of GPI, probably the dimer, was found to be inactive in this bioassay. Using the neuroblastoma cells model system, we further investigated the significance of the region of homology to HIV-1 envelope glycoprotein (gp120) as the putative binding site of GPI to its receptor on neuronal cells. PMID- 2547085 TI - Presence of alpha-melanocyte-stimulating hormone-like immunoreactivity in the innervation of amphibian skeletal muscle. AB - Amphibian motor nerve terminals are sensitive to a wide variety of peptides, including alpha-melanocyte-stimulating hormone (alpha-MSH). We determined the presence and distribution of alpha-MSH-like immunoreactivity (alpha-MSHLI) in the innervation of the cutaneous pectoris muscle from bullfrog (Rana catesbeiana) tadpoles and postmetamorphic froglets, and adult frogs (R. catesbeiana and R. pipiens). alpha-MSHLI was found in unmyelinated, noncholinergic axons, in motor axons, and in motor nerve terminals. In motor axons, alpha-MSHLI was predominantly associated with neurofilaments. The distribution of this form of alpha-MSHLI changed during development and seasonally in adult frogs. The possible functional roles of this alpha-MSHLI are discussed. PMID- 2547087 TI - Infectious mononucleosis and mononucleosis-like illnesses in children and adults in Saudi Arabia. AB - The diagnosis of heterophil antibody positive, heterophil negative infectious mononucleosis and mononucleosis-like illnesses as made in 58 children and adult patients with clinical features suggestive of the mononucleosis syndrome and significant number of atypical lymphocytes. Epstein-Barr virus (EBV) specific serological tests revealed that six children and 23 adults had primary heterophil positive EBV infections based on the detection of IgM to EB-viral capsid antigens (IgM-VCA), presence of IgG to DB-early antigens (EA-D) and absence of antibodies to EB-nuclear antigen (EBNA). A further nine cases (five children, four in adults) of heterophil negative EBV infectious mononucleosis were likewise detected by EBV-specific serologic tests. Fourteen cases including one in children were due to active cytomegalovirus (CMV) infections as evident by positive CMV-IgM; they were all heterophil negative. Of the remaining heterophil negative cases, one was due to T. gondii with positive Toxo-IgM and five cases were of undetermined aetiology. PMID- 2547086 TI - Induction of protooncogene fos by extracellular signals in primary glial cell cultures. AB - In the present study various extracellular factors, acting through different second messenger systems, were examined for their capacity to increase the level of c-fos mRNA in primary glial cell cultures. In particular EGF, 12-O tetradecanoylphorbol 13-acetate, the beta-adrenergic agonist isoproterenol, and the glutamate agonists, ibotenic and quisqualic acid, were studied. All the extracellular stimuli tested induced a rapid and transient increase in c-fos mRNA level in glial cell cultures regardless of the signal transduction pathway and the final effect on cell proliferation. PMID- 2547088 TI - [The regulatory mechanisms of prostacyclin generation in human vascular endothelial cells]. PMID- 2547089 TI - [cDNA structure of human thrombomodulin]. PMID- 2547090 TI - [Metabolism of thrombomodulin]. PMID- 2547091 TI - [Uncommon renal tumor in children]. AB - We experienced six rare cases of solid renal tumor, which include two benign (CMN, MLCN) and four malignant tumors (CCSK 2, MRTK2). CMN is the most common benign solid tumor in the neonate. MLCN has typical US or CT findings manifested by multilocular cysts and thick wall septa. CCSK and MRTK are rare malignant neoplasms, which present characteristic clinical findings such as bone metastasis or intracranial metastasis. It is important for the radiologist to understand those characteristic clinical presentation of these tumors, because differential diagnosis is often impossible just on the basis of imaging. PMID- 2547092 TI - [A case of multiple peripheral hepatic artery aneurysms after chemoembolization]. AB - A case of 57-year-old man with multiple peripheral hepatic artery aneurysms is reported. The patient was admitted with liver dysfunction and detected hepatoma in the right lobe. Chemoembolization was performed for the treatment of hepatoma. After 35 days, reangiography revealed multiple peripheral hepatic artery aneurysms in the region of chemoembolization. The etiology of aneurysms were presumably drug induced arteritis. PMID- 2547093 TI - [Accumulation of 99mTc-phosphorous compounds in primary hepatic tumors associated with hypercalcemia]. AB - In present study, in two cases with hepatocellular carcinoma and cholangioma associated with hypercalcemia, the accumulations of 99mTc labeled phosphorous compound in the primary lesion were demonstrated and its mechanism was discussed. PMID- 2547094 TI - [Pharmacoangiography of liver neoplasms and gallbladder arteries with angiotensin II]. PMID- 2547095 TI - [In vivo and in vitro studies on PGE2 production and function in human hepatocellular carcinoma]. AB - In order to investigate the production of PGE2 and its' function in human hepatocellular carcinoma, the effects of indomethacin and PGE2 on tumor growth were examined using in vivo and in vitro techniques. HH2-6 cells produced PGE2 in the culture media, and the inverse relationship was observed in between the cell proliferation and the culture supernatant PGE2 levels. While in vivo, plasma and tumor tissue PGE2 levels of tumor bearing nude mice were significantly increased for 1 or 2 weeks after tumor inoculation. In the case of which indomethacin was injected daily into the abdominal cavity (4 mg/kg body weight), the elevation of plasma and tissue PGE2 levels was remarkably suppressed, and the latent time of tumor growth was also prolonged. On the other hand, another case of which PGE2 was injected (10 micrograms or 0.1 microgram i.p.) at first 10 days revealed shortened latent time. These results indicate the intimate relation between PGE2 and latent time on tumor growth. Furthermore, histological findings suggest that tumor derived PGE2 might play an important role in tumor angiogenesis. PMID- 2547096 TI - [Radiation therapy of hepatocellular carcinoma]. AB - After the transhepatic arterial therapy, irradiated 11 patients with hepatocellular carcinoma were investigated clinically and pathologically. As the effect to the main tumors except for 3 cases who dropped out because of the side effect of irradiation, partial response (PR) was found in 1 case, minor response (MR) was in 2. There was no change (NC) in 4 and 1 had progressive disease (PD), so the rate of effectiveness was 12.5%. Irradiation to the tumor thrombosis of the portal vein were done for seven cases, 2 of them (28.6%) showed the decrease of the thrombosis or improvement of the blood stream in the portal vein on angiography or histology. AFP decreased in four of seven measurable cases within one month after the irradiation. Side effects were jaundice, fever, appetite loss and so on, which were seen especially in cases of Child's B or C. Liver dysfunction and bone marrow suppression were also observed though not so severe. For the cases received lipiodol injection before radiation, lipiodol collection to the tumor was beneficial for the determination of the irradiated field. PMID- 2547097 TI - [A case of malignant fibrous histiocytoma with primary site in greater omentum]. PMID- 2547098 TI - [Role of polymorphonuclear leukocytes and oxygen-derived free radicals in the formation of gastric mucosal lesions induced by platelet-activating factor]. AB - Gastric hemorrhagic erosions were induced by the administration of platelet activating factor (PAF). Gastric wall blood flow was decreased and thiobarbituric acid (TBA) reactants in the gastric mucosa were increased 10 min after PAF injection. SOD and catalase reduced gastric mucosal lesions and TBA reactants, but had no influence on gastric blood flow. In polymorphonuclear leukocytes (PMN) depleted rats, gastric mucosal lesions and TBA reactants in the gastric mucosa were reduced and gastric blood flow were increased. PAF induced the superoxide production from rat PMN in a dose dependent manner. These results suggest that oxygen-derived free radicals produced by PMN and lipid peroxidation may play an important role in the pathogenesis of gastric mucosal injury induced by PAF. PMID- 2547099 TI - [The hepatic hemodynamic response to intra-arterial infusion of vasoactive agents -Part 2. Blood flow measurements in rats with liver cancer]. AB - The tissue blood flow (TBF) of primary liver cancer and normal live in 54 rats were measured following infusion of vasoactive agents into the celiac axis under continuous recording of blood pressure. Angiotensin II (AT) (1.0 micrograms/kg/min) and Prostaglandin F2 alpha (PGF) (1.0 micrograms/kg/min) were used. The former increased TBF of the tumor and blood pressure and the latter reduced TBF of the tumor with no remarkable change of the blood pressure, but both drugs made little or no influence on TBF of the normal liver. Dibutyryl cyclic AMP (DBcAMP) (2.0 mg/kg/hr) infusion increased TBF of the both tumor and normal liver with no significant change of blood pressure. PGF (1.0 micrograms/kg/min) infusion after preinfusion of DBcAMP did not cause the decrease of TBF of the tumor. PMID- 2547100 TI - [A case of ulcerative colitis with multiple colon cancers]. PMID- 2547101 TI - [A case of liver cirrhosis associated with monoclonal gammopathy and primary hepatoma]. PMID- 2547102 TI - [Ambulatory follow-up of cancer patients undergoing chemotherapy--QOL (quality of life) of cancer patients]. PMID- 2547103 TI - [Errors in the diagnosis of a Barre-Masson tumor]. PMID- 2547104 TI - [A germ cell tumor of an ectopic testis in polyorchidism]. PMID- 2547105 TI - [Protracted forms of viral hepatitis A in adolescents]. AB - The results of study of the clinical picture and outcomes of type A viral hepatitis (HA) in 80 male adolescents aged 15-18 years are presented. The diagnosis was confirmed serologically. A greater incidence of the protracted forms of HA has been found in adolescents which in most cases had a favourable outcome. Protracted hepatitis was marked by long-term persistence of anti-HVA IgM. The chronic form of viral hepatitis had a course of a mixed A and B viral infection. PMID- 2547106 TI - [Lymphotropic glucocorticosteroid treatment of patients with corticosteroid dependent bronchial asthma]. PMID- 2547107 TI - [Cyclic nucleotide levels in the gastric juice of patients with recurrent duodenal ulcer]. AB - Cyclic nucleotide content in the gastric juice of patients with duodenal ulcer recurrences was studied before and after reconstructive surgery and in remote periods. The basal secretion phase was characterized by an increased cyclic nucleotide concentration in gastric juice; histamine and insulin stimulation caused decrease in their concentration. After reconstructive surgery, acid production in the basal period decreased, while cyclic adenosine monophosphate concentration increased by more than 100 per cent. The long-term period was characterized by clear intensification of cyclic adenosine monophosphate excretion under the action of histamine. It has been suggested that the release of this nucleotide from gastric mucosa cells is a subsidiary mechanism for maintenance of its necessary intracellular concentration. PMID- 2547108 TI - [Low and high-molecular dietary bulk products]. PMID- 2547110 TI - Distribution of vasoactive intestinal peptide and its receptors in the arteries of the rabbit. AB - Vasoactive intestinal peptide (VIP) is a widely distributed neurotransmitter whose dilatory effects on vascular smooth muscle are believed to be mediated via specific receptors. To determine the possible role of VIP in regulating specific vascular beds, we examined the relationship between arterial wall VIP content as determined by radioimmunoassay and VIP receptors mapped by autoradiography. Analysis of arteries from 12 adult New Zealand rabbits showed that VIP receptors were consistently located in the wall of all muscular arteries, and that the 125I VIP grain density correlated with VIP content. 125I-VIP binding in the mesenteric, renal, and iliac arteries was abundant and their VIP content was 192 +/- 56, 51 +/- 5, and 74 +/- 23 fmole/mg protein, respectively. 125I-VIP binding to the thoracic aorta was indistinguishable from nonspecific binding, its VIP content being 15 +/- 2 fmole/mg protein. The abundance of VIP receptors and the high VIP levels associated with the mesenteric, renal, and iliac arteries suggest that VIP is a potential regulator of flow to the vascular beds supplied by these arteries. In contrast, the much lower density of receptors in the extracranial carotid, which is also a muscular artery, suggests that, in rabbits, control of carotid vasomotion may be less dependent on VIP innervation. Furthermore, these results suggest that VIP receptors and VIP-containing neurons are not uniformly distributed in the arterial vasculature and that VIP may have selective vasodilatory effects. PMID- 2547109 TI - Correction of sphingomyelinase deficiency in Niemann-Pick type C fibroblasts by removal of lipoprotein fraction from culture media. AB - The average sphingomyelinase activity of fibroblasts obtained from 12 Niemann Pick type C patients was 37.9% of that of normal fibroblasts (27.2 versus 72 nmol (mg protein)-1 h-1) when the cells were cultured in minimum essential medium containing 13% fetal bovine serum. Following replacement of the above medium with medium in which the lipoprotein fraction had been removed from the fetal bovine serum, the sphingomyelinase activity rose over a 7-day period from about 1/3 of normal to normal or above. Upon reintroduction of medium containing 10% fetal bovine serum which had not been extracted, the sphingomyelinase activity of the Niemann-Pick type C cells again fell within 48 h to 30% of the normal controls. In contrast, cell lines from patients with either Niemann-Pick Type A or B were not influenced by the presence or the absence of lipoprotein, i.e. lacked sphingomyelinase activity under all culture conditions examined. Histochemical staining with filipin showed an inverse relationship between the sphingomyelinase activity and intracellular, free, unesterified, cholesterol level. Moreover, immunochemical staining with an antibody against a lysosomal membrane protein provided direct evidence that the accumulation of unesterified cholesterol in cells cultured in regular (non-extracted) medium occurred within lysosomes and/or related organelles. PMID- 2547111 TI - Structural and functional alterations in the gut of parenterally or enterally fed rats. AB - Various regimens of total parenteral nutrition (TPN) and enteral feeding were compared to determine their effects on the structural and functional changes of rat small intestine. Male Wistar rats, allocated randomly into five groups on the basis of delivery route and composition of nutrients, were fed as follows: standard rat chow ad libitum (CE-2 group), low-residue diet (LRD group), LRD supplemented with 1% (w/v) fiber (LRD + fiber group), elemental diet (ED group), and TPN (TPN group). At 2 weeks of feeding, villi in the terminal ileum decreased in height in the following order: CE-2 group greater than LRD + fiber group greater than LRD group greater than ED group greater than TPN group. Mucosal diamine oxidase activity remained unchanged in the CE-2 group and LRD + fiber group throughout the experimental period. However, mucosal diamine oxidase values were significantly lower in the remaining three groups, similar to the structural changes, and those values in the ED group were significantly decreased at 2, 3, and 4 weeks. There was a positive correlation between plasma diamine oxidase level and mucosal diamine oxidase content, with a coefficient correlation of y = 0.20x + 0.03, r = 0.55 (P less than 0.01). These results could be interpreted to indicate that addition of dietary fiber to LRD has a favorable effect on the maintenance of intestinal architecture and function during enteral feeding, and plasma diamine oxidase activity can be used as an index of functional and/or structural changes occurring in the small intestine during enteral or parenteral feedings. PMID- 2547112 TI - The cytotoxic effects of estradiol-17 beta, catecholestradiols and methoxyestradiols on dividing MCF-7 and HeLa cells. AB - In this study the cytotoxic effects of high concentrations (greater than or equal to 1 x 10(-6) M) of estradiol-17 beta (E2), 2-/4-hydroxyestradiol-17 beta (2-/4 OHE2) and 2-/3-/4-methoxyestradiol-17 beta (2-/3-/4-MeOE2) were determined on dividing MCF-7 and HeLa cells. The 2-MeOE2 metabolite followed by 2-OHE2 and E2 (in this order) proved to be extremely toxic to dividing MCF-7 and HeLa cells. The cytotoxic effect on these cells comprised uneven chromosome distribution. Indirect immunofluorescent studies, in which monoclonal anti-alpha-tubulin antibodies were used, showed that these compounds (2-MeOE2 greater than 2-OHE2 greater than E2) at high concentrations caused abnormal and fragmented polar formations as well as disorientated microtubule arrangement in the dividing MCF-7 and HeLa cells. The 4-OHE2 and 3-/4-MeOE2 metabolites had little or no cytotoxic effects on dividing cells. The large number of abnormal metaphases seen in HeLa cells exposed to 2-MeOE2 suggested that this metabolite may be the ultimate cytotoxic compound. The reduction in the number of HeLa cells with abnormal metaphase configurations after exposure to 2-OHE2 plus quinalizarin (an inhibitor of catechol-O-methyltransferase) indicated that the production of 2-MeOE2 is necessary for the formation of abnormal spindles in metaphase. Quinalizarin treatment in the presence of 2-MeOE2 had no effect on the large number of abnormal metaphases. We therefore conclude that neither E2 nor 2-OHE2, but a high concentration of 2-MeOE2 is responsible for abnormal spindle formation. In additional experiments the number of normal and abnormal dividing HeLa cells were greatly reduced when simultaneously exposed to E2 and 2-/4-hydroxylase-inhibitor alpha-naphthoflavone. PMID- 2547113 TI - Progesterone metabolism in the gastric mucosa microsomes of guinea pig. AB - The microsomes from guinea pig gastric mucosa were found to convert [4 14C]progesterone to two major metabolites in the presence of NADPH. The gastric metabolizing activity was the highest among the gastrointestinal tissues of guinea pig. 5 alpha-Pregnane-3,20-dione and 3 beta-hydroxy-5 alpha-pregnan-20-one were identified as the major metabolites by thin-layer chromatography and crystallization to constant specific activity, suggesting the presence of steroid 5 alpha-reductase and 3 beta-hydroxysteroid dehydrogenase activities in the gastric mucosa microsomes. Furthermore, time course of progesterone metabolism and analysis of 5 alpha-pregnane-3,20-dione metabolites suggest that the gastric progesterone metabolism is initiated by 5 alpha-reductase and followed by 3 beta hydroxysteroid dehydrogenase. The progesterone-metabolizing activity was strongly inhibited by SKF 525-A and disulfiram. The activity was also inhibited by methyrapone to a somewhat lesser extent than the above inhibitors. From gastric mucosa microsomes, the progesterone-metabolizing activity was successfully solubilized with 2% digitonin using 0.1 M potassium chloride and 1 mM dithiothreitol, 0.4 mM NADPH and 20% glycerol as stabilizers for the solubilized activity. Among these stabilizers, glycerol was found to be most effective for stabilizing the activity of the solubilized microsomes. PMID- 2547114 TI - Effects of the infusion with ACTH or CRH on the secretory activity of rat adrenal cortex. AB - Six-hour infusion with ACTH or CRH induced a dose-dependent rise in the plasma concentrations of ACTH, corticosterone (B) and aldosterone (A). Positive linear correlations between the plasma levels of ACTH and B or A were found in both ACTH or CRH-infused animals. Regression curves for B were similar in both groups of animals, while the regression line for A was significantly (P less than 0.05) steeper in CRH-than in ACTH-treated rats. These findings suggest that, in the rat, the mechanism underlying the CRH-induced stimulation of A secretion does not exclusively involve the enhancement of ACTH release. PMID- 2547116 TI - Hepatocellular carcinoma: some aspects to improve long-term survival. AB - Sixty-six patients surviving over 5 years after resection of hepatocellular carcinoma (HCC) are investigated. Of them, subclinical stage amounted to 56.1% (37/66) and moderate stage to 43.9% (29/66). There were 35 cases with small HCC (less than or equal to 5 cm). Cirrhosis was present in 81.1% (54/66). Radical resection was performed in 98.5% (65/66) and palliative resection in 1.5% (1/66). Reoperation for subclinical recurrence and solitary pulmonary metastasis was done in 14 patients, and sequential resection of huge tumors, in three patients. By the end of June 1988, follow-up varied from 60 to 319 months (mean, 115 months); 80.3% of the patients (53/66) are still alive and free of disease; 19.7% (13/66) died with disease. The majority of long-term survivors have returned to their original work; some young patients got married after resection of small HCC 10 years ago, and some can even play football again. Some aspects to improve long term survival are discussed. PMID- 2547115 TI - Structural basis of steroid compounds interaction with "digitalis"-receptor sites of Na,K-dependent ATPase. AB - Twenty-two Steroid molecules have been tested for the inhibition Na,K-dependent ATPase at 10(-7)-10(-4) M concentrations. At the 10(-5) M concentration of the investigated molecules, inhibition ranged from 8 to 36%. To explain the structure inhibition % relationship, we determined the value of heteropolarity or biphilicity moment of these molecules. This value would appear to be dependent on the space location and hydrophilicity of the molecule elementary fragments, and to the degree of their water accessibility; however, it is independent of the hydrophilicity of the molecules as a whole. On the basis of the obtained data, details of Na,K-ATPase digitalis-receptor structure and the mechanism of the glycoside-receptor interaction are discussed. PMID- 2547117 TI - Esophageal metastasis from breast carcinoma associated with pseudoepitheliomatous hyperplasia: an unusual endoscopic diagnosis. AB - The clinicopathologic findings in a patient who developed dysphagia and obstruction of the middle third of the esophagus 12 years after mastectomy for breast carcinoma are presented. Endoscopic biopsy revealed metastatic breast carcinoma with pseudoepitheliomatous hyperplasia of the overlying esophageal mucosa. To the best of our knowledge, the case represents the first reported association between esophageal metastasis and pseudoepitheliomatous hyperplasia of the esophageal squamous epithelium. PMID- 2547118 TI - Simplified HPLC method for quantitation of vitamin D in blood serum. AB - A single-step high performance liquid chromatography (HPLC) method for vitamin D quantitation in blood serum is described. Methanol/hexane extraction, silica Sep Pak purification, differential dissolution, filtration, and the use of a precolumn in place of an injector sample loop allow for using only one (reversed phase) HPLC step instead of the usual two. Detection limit with the HPLC system used was about 1 micrograms/L of serum. Recovery of vitamin D3 added to serum ranged from 90% to 109%. Intra and interassay coefficient of variation values were 8.1% and 15.4%, respectively. PMID- 2547119 TI - Simple determination of acetylcholine and choline within 4 min by HPLC-ECD and immobilized enzyme column in mice brain areas. AB - A rapid and simple method using a combination of high-performance liquid chromatography (HPLC) with electrochemical detection (ECD) and an immobilized enzyme column herein is described for simultaneous assay of acetylcholine (ACh) and choline (Ch) in discrete brain areas of the mouse. Perchloric acid extracts of small brain tissues were injected directly onto the HPLC system with no pre cleanup procedure. The ACh and Ch levels were determined within 4 min for each chromatographic run. Recoveries were 100.0 +/- 4.8% for ACh and 96.1 +/- 1.4% (mean +/- S.D., n = 10) for Ch. Detection limits for both compounds were 0.25 pmol (signal-to-noise ratio of three). The present method has been applied to the measurement of Ch and ACh in several discrete brain areas of the mouse. PMID- 2547120 TI - Evaluation of a semiautomatic cell harvester filtration for the determination of beta-adrenoceptors in human mononuclear leukocytes. AB - The ligand affinity (Kd) and the number (Bmax) of beta-adrenoceptors in human mononuclear leukocytes was measured using a semiautomatic cell harvester method for the separation of bound and free ligand. The method was first evaluated in terms of contamination, nonspecific filter binding, and well-to-well reproducibility. A comparison was then made with the conventional manual filtration method. The Kd values for the binding of [125lodo]-cyanopindolol to mononuclear leukocyte membranes were independent of the method, whereas the Bmax values were systematically higher (around 30%) when determined with the cell harvester filtration method. A highly significant correlation was found between the Bmax values obtained with both methods. PMID- 2547122 TI - [Ascites and renal insufficiency in a 59-year-old male subjected to liver transplant for hepatocarcinoma on liver cirrhosis]. PMID- 2547121 TI - A microtest plate assay of functional excitatory amino acid receptors. AB - A microtest plate assay of functional excitatory amino acid receptors present on cultured chick embryo retinal cells has been developed. It is based on measurements of excitatory amino acid-mediated increase in 22Na+ influx into retinal cells adhering to each of the 96 wells of microtest plates. Dose dependent responses to L-glutamate, kainate and N-methyl-D-aspartate but not to quisqualate can be measured. These responses are selectively inhibited by antagonists of excitatory amino acids. The assay is reliable, fast to perform and parsimonious in terms of the volume and thus of amount of the drug applied. It allows a single investigator to perform, in one day, measurements of the effects of known or putative glutamatergic ligands in more than 100 different conditions. PMID- 2547123 TI - [Neuroendocrine effects of buspirone]. PMID- 2547124 TI - Applications of a microcomputer-based reconstruction system to produce realistic three-dimensional shaded images from serial sections. AB - A microcomputer-based system is described that generates three-dimensional shaded images from serial section data. It is implemented on the IBM PC-AT microcomputer, using a graphics display card (Real World Graphics PC 4000) to provide sufficient colours for the production of realistically shaded and highlighted images. Image generation time is typically less than 10 min. The capabilities of the system are illustrated by reconstructions of the Kurloff cell from the guinea-pig (based on serial electron micrographs), human teeth (based on light microscope data), the human leg (from gross anatomical sections) and the human brainstem (from stereotaxic atlas data). The system is applicable to a wide variety of serial section data from histological, anatomical and medical imaging systems and ultrastructural studies. PMID- 2547125 TI - Recent developments in the design of receptor specific opioid peptides. PMID- 2547126 TI - Mode of action of razoxane: inhibition of basement membrane collagen-degradation by a malignant tumor enzyme. AB - Razoxane is an antitumor agent structurally related to EDTA. Its cytotoxic mechanism of action has been shown to involve interference with cell division and reduction of the gross rate of DNA synthesis. In this study we present evidence that razoxane, at therapeutically attainable concentrations, inhibits type IV collagen and basement membrane degradation by an enzyme isolated from malignant tumors. This effect is attributed to the chelating properties of razoxane and may explain the antimetastatic properties, by means of stabilization of tumor blood vessels, which have been associated with the action of the antitumor drug. PMID- 2547128 TI - Effect of different stimulators and a blocker of insulin release on islet Na+, K+ ATPase activity. AB - The effect of several insulin secretagogues and a blocker upon islet Na+, K+ ATPase activity was studied using rat islet homogenates. None of the agents tested modified the enzyme activity when added directly to the enzyme assay. Activity of Na+, K+-ATPase measured in islets preincubated during 3 min with glucose 3.3, 8 or 16.6 mM, as well as with 15 mM KIC or 1.2 microM somatostatin, did not significantly change. The presence of glucagon (1.4 microM) plus theophylline (10 mM) in the preincubation medium significantly enhanced activity while tolbutamide (1.48 mM) or gliclazide (76 microM) significantly decreased such activity. These results suggest that Na+, K+-ATPase activity would not be a main common step involved in the mechanism by which glucose, KIC, glucagon + theophylline and somatostatin exert their effect on insulin secretion. Conversely, the enzyme might contribute to the stimulatory effect of gliclazide and tolbutamide on insulin release. Such effect would be secondary to the release of some cellular mediator rather than a direct action of these compounds on the enzyme. Such effect would later favor a rise in the cytosolic concentration of calcium which might trigger the release of insulin. PMID- 2547127 TI - Beta-adrenoceptor binding potencies of new aliphatic and alicyclic oxime ethers and their relevance to intraocular pressure control. AB - The selectivity and binding potency of a series of alkyliminoxypropanolamines characterized by the lack of an aromatic nucleus are studied using a new non selective radioligand, (-)-(125I)-Iodocyanopindolol. The relationship between the effectiveness in lowering the intraocular pressure (IOP) in experimental hypertensive rabbit eyes and their capacity to bind to ciliary processes beta 2 adrenoceptors is discussed. The inhibition constant (Ki) and beta 2/beta 1 ratios indicate a beta 2-selectivity for the tested drugs. Cyclopropyl, dicyclopropyl, cyclopentyl and cyclohexyl derivatives displayed a potent binding to ciliary processes beta 2-adrenoceptors and lowered the IOP about -18%. These compounds induced a lowering in IOP equal to that produced by timolol and appear to be effective and safe beta-adrenergic antagonists in open-angle glaucoma therapy. Decreasing the size of the alkyl group of the oxime, removing the oxime function or modifying the beta-hydroxyl group from the side chain led to a significant decrease in beta 2-adrenoceptor binding and induced weak hypotensive ocular activity. Since the tested alkyliminoxypropanolamine series has very similar physicochemical characteristics and therefore, ruled out the differences in their ability to reach, through the cornea, the targeted ciliary processes, it was demonstrated that contrary to generally held views, the action of the new beta antagonist series on IOP is related to their ability to antagonize ocular beta 2 adrenoceptors. PMID- 2547129 TI - BRL 43694, a new 5HT3 receptor antagonist, prevents cisplatin-induced emesis in dogs. PMID- 2547130 TI - Thiobarbituric acid-reactive malondialdehyde formation during superoxide dependent, iron-catalyzed lipid peroxidation: influence of peroxidation conditions. AB - A systematic study of the influence of biological lipid peroxidation conditions on lipid hydroperoxide decomposition to thiobarbituric acid-reactive malondialdehyde is presented. A superoxide-dependent, iron-catalyzed peroxidation system was employed with xanthine oxidase plus hypoxanthine plus ferric iron adenosine diphosphate complex as free radical generator. Purified cardiac membrane phospholipid (as liposomes) was the peroxidative target, and 15 hydroperoxy-eicosatetraenoic acid was used as a standard lipid hydroperoxide. Exposure of myocardial phospholipid to free radical generator at physiological pH (7.4) and temperature (37 degrees C) was found to support not only phospholipid peroxidation, but also rapid lipid hydroperoxide breakdown and consequent malondialdehyde formation during peroxidation. Under lipid peroxidation conditions, oxidative injury to the phospholipid polyunsaturated fatty acids required superoxide radical and ferric iron-adenosine diphosphate complex, whereas 37 degrees C temperature and trace iron were sufficient for lipid hydroperoxide decomposition to malondialdehyde. Harsh thiobarbituric acid-test conditions following peroxidation were not mandatory for either lipid hydroperoxide breakdown or thiobarbituric acid-reactive malondialdehyde formation. However, hydroperoxide decomposition that had begun in the peroxidation reaction could be completed during a subsequent thiobarbituric acid test in which no lipid autoxidation took place. Iron was more critical than heat in promoting the observed hydroperoxide decomposition to malondialdehyde during the lipid peroxidation reaction at 37 degrees C and pH 7.4. These data demonstrate that the radical generator, at physiological pH and temperature, serves a dual role as both initiator of membrane phospholipid peroxidation and promotor of lipid peroxide breakdown and thiobarbituric acid-reactive malondialdehyde formation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547131 TI - Bradykinin-increased phospholipid deacylation-reacylation in rat renal medulla is inhibited by dBc AMP. AB - The effect of bradykinin on the mobilization of arachidonic acid was analyzed separately by acylation and degradation. Acylating activity was determined by the incorporation of [14C]arachidonic acid into the phospholipids at different times. Different concentrations of bradykinin had no effect on the phospholipid acylating activities. The degradation of the phospholipids was performed on renal medullary slices prelabeled with [14C]arachidonic acid. Treatment with bradykinin produced an initial degradation of phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol, with a concomitant increase in lysophosphatidylcholine, lysophosphatidylethanolamine and lysophosphatidylinositol within 5 min of incubation. Phosphatidylcholine-, phosphatidylethanolamine- and phosphatidylinositol increased thereafter and reached the control values after 10 min of incubation. After 30 min, incubation of prelabeled slices with bradykinin produced a significant concentration-dependent increase in the phospholipid labeling by reutilization of [14C]arachidonic acid. The effect of bradykinin on the phospholipid-labeling was blocked by preincubation with increasing concentrations of dBc AMP. Mepacrine also blocked the bradykinin stimulation in phosphatidylcholine and phosphatidylethanolamine, but had no effect on bradykinin induced changes in the phosphatidylinositol arachidonic acid moiety. PMID- 2547132 TI - Three new and bioactive icosanoids from the temperate red marine alga Farlowia mollis. AB - Three new dihydroxyicosanoids, 12(R),13(R)-dihydroxyicosa-5(Z),8(Z),10(E),14(Z) tetraenoic acid, 12(R),13(R)-dihydroxyicosa-5(Z),8(Z),10(E),14(Z),17(Z)-pentaeno ic acid and 10(R*),11(R*)-dihydroxyoctadeca-6(Z),8(E),12(Z)-trienoic acid, have been isolated from a previously unstudied temperate red marine alga, Farlowia mollis (Cryptonemiales, Rhodophyta). The structures of these new metabolites have been deduced from detailed nuclear magnetic resonance and mass spectrometry analyses on stabilized diacetate-methyl esters and stereochemistry deduced by 1H NMR couplings and CD analysis of a dibenzoate derivative. Collectively, these new natural products modulate fMLP-induced superoxide anion generation in human neutrophils, inhibit the conversion of arachidonic acid to lipoxygenase products by human neutrophils, and inhibit the functioning of the dog kidney Na+/K+ ATPase. PMID- 2547133 TI - Eicosapentaenoic and arachidonic acid: comparison of metabolism and activity in murine epidermal cells. AB - The biological activity, including metabolism and modulation of ornithine decarboxylase activity and DNA synthesis, of arachidonic acid (AA) and eicosapentaenoic acid (EPA) were compared in epidermal cells from SENCAR mice. Radiolabelled AA and EPA were found to be similarly incorporated into and released from membrane phospholipids of unstimulated cultures. However, when cells were stimulated with the tumor promoter 12-0-tetradecanoylphorbol-13 acetate (TPA), the release of AA was significantly higher than the release of EPA. The extent of metabolism of AA and EPA to prostaglandins was determined in both freeze-thawed cell preparations and in viable cultured cells. In the freeze thawed preparations, use of AA as a substrate resulted in significantly more PGF than when EPA was used as the substrate. However, more PGE3 was formed than PGE2. PGD levels were the same for either fatty acid precursor. Prostaglandin production was also determined in viable cultured cells since other influences such as phospholipase A2 activity can modify prostaglandin production. Control cultures prelabelled with either AA or EPA produced similar amounts of the respective PGF, PGE, and PGD. However, TPA-stimulated cultures produced significantly higher amounts of each prostaglandin in cultures prelabelled with AA compared to cells prelabelled with EPA. HETE or HEPE production was the same both for cultured cells prelabelled with AA or EPA and for homogenates from uncultured cells incubated directly with the radiolabelled fatty acids. TPA induced ornithine decarboxylase (ODC) was significantly higher in AA-treated cultures compared to EPA-treated cultures. AA supports DNA synthesis to a greater extent than EPA, either alone or in the presence of TPA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547134 TI - Therapeutic response of breast carcinoma monitored by 31P MRS in situ. AB - In situ phosphorous MRS was employed to study the metabolites of normal and cancerous breasts, and the alterations of tumor response to therapy. In a group of 7 normal volunteers and 12 patients, the total mobile phosphate content of breast carcinomas was found to be at least two to three times higher than that of the normal breast measured off menstruation. The metabolite profiles of normal and tumorous breasts are coarsely similar. In both cases the intracellular pH (pHi) was either neutral or slightly alkaline (pH greater than 7.0). Prior to treatments, the metabolite levels of phosphoryl monoester-to-ATP ratio of breast neoplasms were higher than those of the controls and decreased after the patients received a few treatments while the pHi fluctuated at a value greater than 7.0. The phosphoryl diester-to-ATP ratio also demonstrated to a lesser extent a decreasing trend in response to therapy. PMID- 2547135 TI - Magnetization transfer contrast (MTC) and tissue water proton relaxation in vivo. AB - In this study the exchange between 1H magnetization in "free" water (1Hf) and that in a pool with restricted motion (1Hr) was observed in tissues in vivo using NMR saturation transfer methods. Exchange between these two pools was demonstrated by a decrease in the steady-state magnetization and relaxation times of 1Hf with radiofrequency irradiation of 1Hr. The pseudo-first-order rate constant for the movement of magnetization from 1Hf to 1Hr was approximately 1 s 1 in kidney and approximately 3 s-1 in skeletal muscle in vivo. Proton NMR imaging demonstrated that this exchange was tissue specific and generated a novel form of NMR image contrast. The extent of exchange between 1Hf and 1Hr as well as the topological correlation of the exchange with relaxation weighted images suggests that this pathway is a major determinant of the observed relaxation properties of water 1H in vivo. PMID- 2547136 TI - [Volatile ashes and their biological effect. 2. Fibrogenic effect of volatile ashes]. AB - In experiments on white Wistar rats fibrogenic effects of 6 samples of fly-ashes collected from electric precipitators in power engineering plants have been evaluated. The coal came from different national deposits. All the ashes have been found to contain: quartz and mullite, 3 ashes contained additionally orthoclase, whereas 1, apart from quartz and mullite, contained kaolinite; naturally radioactive elements (Ra226, K40, Th228) and trace elements (As, Ba, Be, Cd, Ce, Cu, Fe, Pa, Mo, Ni, Pb, Se, U Zu). Experimental pneumoconiosis was induced through intratracheal administration of single doses of 50 mg of dust; the experiment was carried out at 3 time intervals of 3, 6 and 9 months. The fibrogenic activity was evaluated both qualitatively (histopathological methods) and quantitatively (lung weight, hydroxyproline content in lungs, dust elimination from lungs); control groups consisted of animals which obtained NaCl solution and quartz sands. Fly-ashes were found to exhibit different fibrogenic effects, yet, their fibrogenic activity was weaker, compared to quartz sands. No clear correlation was found between fibrogenic effects of ashes and test physico chemical properties, such as the content of SiO2, trace elements or naturally radioactive elements. Analysis of occupational diseases (for the period section): (1979-1983) demonstrated occupational diseases of dust-related aetiology among power engineering workers, pneumoconioses, constituting 7.8% of 127 cases of occupational diseases. PMID- 2547137 TI - Long-term and withdrawal effects of two different oestrogen-progestogen combinations on lipid and lipoprotein profiles in post-menopausal women. AB - Serum lipids and lipoproteins were studied in 86 healthy post-menopausal women at 3-monthly intervals throughout 2 yr of treatment with cyclic oestradiol valerate plus cyproterone acetate (E2V + CPA), continuous 17 beta-oestradiol plus norethisterone acetate (E2 + NETA), or placebo. Withdrawal effects were also investigated. Serum oestrogen levels were similar following the 2 hormone regimens. Serum total and low-density-lipoprotein (LDL) cholesterol were significantly reduced by 5-8% during cyclic E2V + CPA therapy and by 15-20% during continuous E2 + NETA treatment. The concentrations remained significantly reduced throughout the 2 yr of treatment. Virtually no changes were observed in the placebo group. High-density-lipoprotein (HDL) cholesterol concentrations were significantly reduced in relation to pretreatment values during the first year of continuous E2 + NETA therapy, but were not significantly different from the levels in the cyclic E2V + CPA or the placebo groups. Serum triglyceride levels remained almost constant in all the groups. Withdrawal of hormone therapy resulted in steep increases in total and LDL-cholesterol, the levels returning to pretreatment values. A similar increase in HDL-cholesterol was observed in the E2 + NETA group, following withdrawal. Vaginal bleeding episodes were experienced by all the women receiving the E2V + CPA regimen and they occurred regularly in 63% of the women. Fifty-two (52) percent of the women receiving E2 + NETA did not bleed at all. The results of the present study suggest that the continuous addition of NETA may enhance the oestrogen-induced changes in total and LDL cholesterol, whereas only minor changes in the oestrogen-induced effects on these variables are produced by the cyclic addition of CPA. However, in the case of HDL cholesterol, the antagonistic effect of CPA seems to be moderate in comparison with that of NETA. PMID- 2547138 TI - Effect of orally administered oestrogens on gonadotrophin levels in post menopausal women. AB - The gonadotrophin-suppressing effect of 4 peroral oestrogen regimens (A: 2.5 mg piperazine oestrone sulphate; B: 1.25 mg piperazine oestrone sulphate + 5.0 mg oestriol; C: 2.0 mg oestradiol valerate; D: 10.0 mg oestriol; all administered daily) was studied in 7 post-menopausal women, in the absence, and then in the presence of daily oral doses of 250 micrograms of levonorgestrel. A randomized, complete cross-over design was used, and from each volunteer 80 blood samples were drawn during a period of 252 days for the assay of immunoreactive follicle stimulating hormone (FSH), luteinizing hormone (LH) and bioactive LH levels. All 4 formulations significantly diminished pretreatment gonadotrophin levels. Combination with levonorgestrel enhanced the suppressive effects. Subsequent placebo administration for a week restored gonadotrophin levels to those found previously during oestrogen administration, but not to pretreatment levels. Regimen D exhibited the relatively weakest and regimens A and B the strongest suppression, both in the absence and in the presence of levonorgestrel. Treatment with all oestrogen formulations decreased, and the addition of levonorgestrel increased the ratio of bioactive (B) to immunoreactive (I) LH. Seven days of placebo administration abolished the effect of levonorgestrel, but not that of the various oestrogens on the B/I ratios. PMID- 2547139 TI - Effect of orally administered oestrogens on circulating oestrogen profiles in post-menopausal women. AB - The effect of 4 peroral oestrogen regimens without and with levonorgestrel (LNG) supplementation (250 micrograms/day) was studied in 7 post-menopausal women. The regimens were: A: 2.5 mg piperazine oestrone sulphate/day; B: 1.25 mg piperazine oestrone sulphate + 5.0 mg oestriol/day; C: 2.0 mg oestradiol valerate/day and D: 10.0 mg oestriol/day. A randomized complete cross-over design was employed and systemic blood levels of progesterone (P), oestrone (E1), oestradiol (E2), oestriol (E3), oestrone sulphate (E1S), oestradiol sulphate (E2S) and oestriol sulphate (E3S) were analyzed. All subjects were consistently post-menopausal, as indicated by low P, E2, E1S and E3S and high FSH and LH levels. The pretreatment and post-treatment levels of E2 and E2S as well as those of E3 and E3S were invariably below detection limit (60 pmol/l and 220 pmol/l, respectively). Treatment with regimens A, B and C resulted in consistently detectable levels of E2, E2S and E3S, but not of E3, and in grossly elevated E1 and E1S levels. Treatment with regimen D gave rise to very high E3S levels. The oestrogen profiles produced by regimens A and C were almost identical, with the exception of higher E1 and lower E3S levels following the administration of regimen A. Combination with LNG did not modify the levels of unconjugated oestrogens, but in certain cases it seemed to diminish those of oestrogen sulphates. A comparison with the results of a preceding study (Aedo A-R et al., Effect of orally administered oestrogens on gonadotrophins levels in post-menopausal women. Maturitas 1989; 11:) reveals that the gonadotrophin-suppressing effect of all 4 regimens still persisted in the post-treatment period at a time when the levels of all oestrogens analyzed returned to those of the pretreatment period. PMID- 2547140 TI - Effect of rice fiber on human fecal microflora. AB - The effects of brown rice, containing fourfold as much dietary fiber as polished rice, on the human fecal microflora were determined. Significantly increased numbers of Bifidobacterium adolescentis and Enterococcus faecalis were observed during the brown rice intake, whereas the total counts and the numbers of bacteroides, Eubacterium aerofaciens, and Escherichia coli during the intake were lower than those before and after the intake. Lower numbers of clostridia and low incidences of Clostridium paraputrificum and C. perfringens were observed during the brown rice intake. PMID- 2547141 TI - Human infection with encephalomyocarditis virus in New South Wales. PMID- 2547142 TI - [Comparative efficacy of tiniba and fasigyn in treating primary and persisting Giardia infections]. AB - Efficacity of tiniba (tinidasole, Kadila, India) and of fasigyn (Polfa, Poland) dosed 300 mg daily for 7 days was 87.5 and 85.2% (P greater than 0.05) immediately after treatment and 45.5 and 50.0% (P greater than 0.05) 4-8 months later, respectively. The lack of parasitocidal effect was established in 4 out of 27 tiniba-treated patients and in 3 out of 24 fasigyn treated ones. Direct correlation between the efficacy of persisting giardia infection chemotherapy and the cAMP phosphodiesterase activity in blood lymphocytes was found previously. Authors established an indirect correlation between the former and hydrochloric acid secretion by the bowel. PMID- 2547143 TI - [The significance of platelet activating factor and its antagonists in bronchial asthma]. PMID- 2547144 TI - [Malignant fibrous histiocytoma of the lung]. PMID- 2547145 TI - Loss of muscarinic cholinergic receptors from the temporal cortex of alcohol abusers. AB - It is currently controversial whether all the brain damage in alcohol abusers in the result of thiamine deficiency (Wernicke-Korsakoff's disease) or whether, in addition, alcohol abuse may affect the brain by other mechanisms as well. The purpose of this study was to determine if alcohol abuse affects muscarinic cholinergic and benzodiazepine receptors in histologically normal brains obtained at autopsy in a general hospital population. Patients were excluded from this study if they had clinical brain diseases (including Wernicke's disease), died in coma, or had liver disease, significant brain atrophy, or dementia severe enough to require institutionalization. We found that muscarinic cholinergic synaptic receptor density determined with [3H] quinuclidinly benzilate was decreased by 40% in homogenates of the tempeoral cortex of 26 alcohol abusers compared with 26 matched nonalcoholic controls. The affinities of the muscarinic receptors were not significantly different between the two groups. In contrast, receptor densities and affinities of benzodiazepine receptors determined with [3H]flunitrazepam were not significantly different in the two groups. Age and death-autopsy time interval had no significant effects on either wet tissue protein concentrations, yields of protein after centrifugation, or receptor binding. The contributions of age and time interval were each less than 2% of the total variance of protein concentrations and receptor binding. When patients were excluded or included who had received cholinergic, anticholinergic, or benzodiazepine medications before death, no significant effects on the final results were observed. Pneumonia, known to be associated with acute hypoxia, and chronic obstructive pulmonary disease, known to be associated with chronic hypoxia, where approximately equally distributed between the two groups and had no significant effects on the results reported here. The loss of muscarinic and the sparing of benzodiazepine receptors occurs in the temporal cortex of histologically normal brains in the absence of significant atrophy and of gross dementia. This means that these changes are early in the development of an alcohol encephalopathy. We have previously reported a decrease in both muscarinic and benzodiazepine receptor binding in the frontal cortex and a decreasing muscarinic but not benzodiazepine receptors in the temporal cortex of alcohol abusers. Taken together, these findings suggest that alcohol neurotoxicity does not simply result in a random loss of neurons and or their associated synapses with their receptors. Instead, different types of receptors, depending upon their location in different brain regions, are specifically affected or spared. This suggests the involvement of region- and receptor-specific mechanisms in chronic alcohol toxicity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2547146 TI - Gangliosides in SV-40-transformed cells derived from Tay-Sachs disease fetal brain. AB - A human glial brain cell line derived from a Tay-Sachs disease fetal cerebellum was transformed with SV-40 virus in order to obtain a transformed brain cell line which reflected the characteristics of the disease. It was shown that the transformed TSD cell line maintained an elevated level of GM2 which was similar to that shown by the nontransformed precursor. In addition, the TSD transformed line lacked hexosaminidase A. PMID- 2547149 TI - Hospital antitrust liability in dealings with downstream providers. AB - As hospitals continue to offer an increasing number of post-discharge services to their patients, the risk of violating antitrust laws also increases. The author, an attorney, discusses the legal complexities of deciding when, if at all, an antitrust law may be violated. PMID- 2547148 TI - Effect of the radioprotector WR-2721 on operant behavior in the rat. AB - The effect of WR-2721 on performance maintained by a fixed-ratio 20 (FR-20) schedule for water reinforcement was studied in male Sprague-Dawley rats. Graded doses of WR-2721 (range 25-100 mg/kg) were administered IP immediately prior to a 60 min test session. WR-2721 had a dose dependent monotonic disruptive effect on FR responding, with significant effects at doses of 50, 75 and 100 mg/kg. WR-2721 also decreased postsession water consumption, but only one significant effect at the highest dose (100 mg/kg). Both slopes of the dose-response regression line are parallel in effect. These data indicate that WR-2721 may affect drinking motivation, which could disrupt operant performance, and WR-2721 affects motor behavior at lower doses than those that depress "motivation" to drink. The log dose-probit analysis on the all-or-none disruptive pattern of pause of responding observed from cumulative records disclosed that the slope of this regression line (s = 1.11) was also almost identical to that of reinforcer decrement analyzed from graded dose-response relationship (s = 1.14) and shared the same estimated ED50's (58.5 and 55.6 mg/kg, respectively). A preliminary study using a variety of pharmacological interventions was also carried out to ascertain if the general functional gastrointestinal disorders produced by WR-2721 may subserve the behavioral deficits. Subcutaneous pretreatments with various selective, peripherally active, gastroprotective drugs [cimetidine (30 and 60 mg/kg), pirenzepine (5 and 10 mg/kg) and domperidone (1, 5 and 10 mg/kg)] 30 min prior to challenge with WR-2721 at dose of 100 mg/kg, demonstrated that these drugs did not yield any apparent significant attenuative effects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547150 TI - Progress toward eradicating poliomyelitis from the Americas. PMID- 2547147 TI - The ras oncogene--an important regulatory element in lower eucaryotic organisms. AB - The ras proto-oncogene in mammalian cells encodes a 21-kilodalton guanosine triphosphate (GTP)-binding protein. This gene is frequently activated in human cancer. As one approach toward understanding the mechanisms of cellular transformation by ras, the function of this gene in lower eucaryotic organisms has been studied. In the yeast Saccharomyces cerevisiae, the RAS gene products serve as essential function by regulating cyclic adenosine monophosphate metabolism. Stimulation of adenylyl cyclase is dependent not only on RAS protein complexed to GTP, but also on the CDC25 and IRA gene products, which appear to control the RAS GTP-guanosine diphosphate cycle. Although analysis of RAS biochemistry in S. cerevisiae has identified mechanisms central to RAS action, RAS regulation of adenylyl cyclase appears to be strictly limited to this particular organism. In Schizosaccharomyces pombe, Dictyostelium discoideum, and Drosophila melanogaster, ras-encoded proteins are not involved with regulation of adenylyl cyclase, similar to what is observed in mammalian cells. However, the ras gene product in these other lower eucaryotes is clearly required for appropriate responses to extracellular signals such as mating factors and chemoattractants and for normal growth and development of the organism. The identification of other GTP-binding proteins in S. cerevisiae with distinct yet essential functions underscores the fundamental importance of G-protein regulatory processes in normal cell physiology. PMID- 2547151 TI - Bistability and control for ATP synthase and adenylate cyclase is obtained by the removal of substrate inhibition. AB - The thesis of this article is that the raison d'etre of the electron transfer chain and the receptor system is to remove 'substrate inhibition' of the enzymes ATP synthase and adenylate cyclase respectively. Activation by energization or hormone is analogous and presents the features of ideal control system; bistability, hysteresis, sensitivity and amplification, and rapid transitions between resting and active states. In the first part of the article, the simplest nontrivial model conforming with the experimental results is put forward. After the system is described, nonlinear and linear models are developed. An important aspect captured by the model is that the enzyme is structurally asymmetric corresponding to the assumption of regulatory site(s) distinct from catalytic site(s). The structural distinction between a regulatory site and a catalytic site entails different binding and specificity properties of the two types of sites with respect to the nucleotides. In the second part, the experimental evidence for the theory is discussed. It is shown that energization and hormone indeed reduce 'substrate inhibition' and that the properties of time lag and criticality predicted by the theory are indeed verified in experiment and are in turn explained by the theory. The theory can explain and correlate various hitherto unexplained experimental phenomena such as the irreversibility of ATP synthesis and the functional role of the ATP synthase asymmetry. The property of hysteresis predicted by the nonlinear model, is indicated by postillumination ATP synthesis, and preactivation of chloroplasts with reduced dithiols indeed display 'hysteresis loops'. In Aplysia memory for short term sensitization may reside in the hysteretic prolonged elevation of cAMP in sensory neurons. PMID- 2547153 TI - Separation of the complexes formed between the regulatory and catalytic subunits of cyclic adenosine monophosphate-dependent protein kinase and topoisomerase I activity in preovulatory follicle-enriched immature rat ovaries. AB - Our previous studies have shown that the regulatory subunits of the type II form of cAMP-dependent protein kinase (RII) present in soluble extract of immature rat ovaries elute from diethylaminoethyl-cellulose as three separate peaks of activity, based on their association with the catalytic subunit (C) of this enzyme, as R2IIC2, an apparent R2IIC, and R2II. Based upon the existence of ovarian RII in three different subunit arrangements, the large amount of C subunit-free R2II in this tissue, and a previous report which indicated that RII exhibited intrinsic topoisomerase I activity, we determined whether DNA topoisomerase I activity was associated with any of these molecular complexes of the ovarian RII subunits. Cyclic AMP-binding activities in soluble extracts of preovulatory follicle-enriched immature rat ovaries were separated by diethylaminoethyl-cellulose chromatography and sucrose density gradient centrifugation. Topoisomerase I activity cosedimented with the apparent R2IIC and R2II obtained from sucrose gradients but was not detected in fractions containing R2IIC2. Upon cAMP affinity purification of the RII derived from fractions containing R2IIC2, R2IIC, and R2II, respectively, no topoisomerase I activity could be detected in any fraction. Phosphorylation of the affinity purified RIIs by the C subunit of beef heart cAMP-dependent protein kinase did not alter this result. These data indicate that none of the RII subunits in soluble extracts of preovulatory follicle-enriched ovaries exhibit intrinsic topoisomerase I activity. PMID- 2547152 TI - Molecular comparison of alpha 2-adrenergic receptors from rat adrenocortical carcinoma and human blood platelet. AB - We have previously described a simple two-step purification technique to isolate alpha 2-adrenergic receptors from the rat adrenocortical carcinoma (Jaiswal, R. K. and Sharma, R. K. (1985) Biochem. Biophys. Res. Commun. 130, 58-64). Utilizing this technique we have now achieved approximately 77,000-fold purification to apparent homogeneity of alpha 2-adrenergic receptors from human platelets. We have compared the biochemical characteristics of these receptors with those from the rat, which were purified approximately 40,000-fold to homogeneity. The [125I] receptor proteins from two sources showed: (a) a single radioactive band with a Mr of 64,000 as evidenced by one- and two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE); and (b) a single symmetrical peak with a pI of 4.2 by isoelectric focusing polyacrylamide gel electrophoresis. Both proteins showed typical alpha 2-adrenergic binding characteristics with specific binding activities of 13.85 nmol/mg and 14.17 nmol/mg protein. These values are close to the theoretical binding activity of 15.6 nmol/mg protein for 1 mol of the ligand binding 1 mol of the receptor protein. These results attest to the purity of the receptors, to its Mr of 64,000, and to its acidic nature. However, the peptide maps of the radioiodinated alpha 2-adrenergic receptors from rat adrenocortical carcinoma and human blood platelets reveal some distinct differences which may relate to the differences in the pharmacological specificities between rodent and non-rodent alpha 2-adrenergic receptors. PMID- 2547155 TI - Induction of human growth hormone promoter activity by the adenosine 3',5' monophosphate pathway involves a novel responsive element. AB - Regulation of GH gene expression by GRF involves cAMP as a second messenger. We have demonstrated that a 500-basepair fragment of the human GH (hGH) gene 5' flanking region can confer cAMP inducibility upon the chloramphenicol acetyltransferase transcription unit in transient transfections of rat pituitary tumor cells treated with forskolin, an activator of adenyl cyclase. The same hGH construct is not induced by forskolin in nonpituitary-derived cells. Experiments with hGH deletion constructs reveal that binding sites for transcription factor AP-2 and the pituitary-specific factor GHF-1 are not required for forskolin stimulation, but that GHF-1 may potentiate the effect. RNA analyses reveal that forskolin also stimulates accumulation of transcripts initiated at the hGH promoter. Other agents that elevate cAMP levels also stimulate hGH expression. Since the hGH 5' flanking region contains no sequences homologous to the cAMP responsive element of the somatostatin gene, and the AP-2 sites do not appear to be required for the forskolin response, these results suggest that a novel cAMP responsive element exists within 82 basepairs upstream from the transcriptional start of the hGH gene and that hGH regulation by GRF may involve interaction between a tissue-specific element and a cAMP-inducible element. PMID- 2547154 TI - Extranuclear estrogen-regulated destabilization of Xenopus laevis serum albumin mRNA. AB - The present study examined 1) whether the estrogen-regulated destabilization of albumin mRNA occurs in the nuclear or extranuclear fraction of the liver cell, and 2) whether the selective posttranscriptional regulation of albumin mRNA stability might result from covalent changes introduced in the processing or polyadenylation of the primary transcript. The disappearance of albumin mRNA after estrogen is restricted to the extranuclear fraction of the cell. Transient changes in steady state levels of the mature nuclear transcript were observed that mirrored the transient estrogen-induced changes previously reported for albumin gene transcription. When assayed 24 h after estrogen (when albumin RNA is virtually undetectable in the extranuclear fraction) the steady state levels of both the primary and mature albumin transcripts found in the nucleus were the same as observed in control animals. Estrogen had no effect on the splicing or selection of polyadenylation sites on the 3'-UTR as determined by high resolution gel analysis of the 3'-UTR and DNA sequencing of cDNA clones isolated from a liver library from an estrogen-treated male Xenopus. Most eukaryotic mRNAs have poly(A) tracts several hundred residues in length, and recent studies have demonstrated that a change in the stability of a number of mRNAs correlates directly with the degree of polyadenylation. Albumin contrasts sharply with this, first because it has an exceptionally short poly(A) tail of 17 residues, and second because the degree of polyadenylation is totally unrelated to its destabilization in response to estrogen. These findings indicate that a unique pathway is involved in the regulation of albumin RNA stability by estrogen in Xenopus. PMID- 2547156 TI - Analysis of rat prolactin promoter sequences that mediate pituitary-specific and 3',5'-cyclic adenosine monophosphate-regulated gene expression in vivo. AB - To determine the rat PRL (rPRL) promoter sequences that mediate pituitary specific and cAMP-induced gene expression in vivo, various lengths of the rPRL promoter were ligated to the luciferase reporter gene and introduced into pituitary and non-pituitary cell lines. A 30-fold increase in rPRL promoter activity was observed in GH4 rat pituitary tumor cells compared to nonpituitary Rat2 fibroblast and HeLa cervical carcinoma cells. About 45% of this cell specific promoter activity was competed by a plasmid containing the -67 to -45 rPRL promoter region, which is the most proximal binding site for a lactotroph specific factor. Compared to a -425 rPRL construct, transfection with rPRL 5'-end points of -212, -178, and -127 contained 23%, 45%, and 1%, respectively, of luciferase activity. Forskolin stimulation resulted in a 10-fold induction of all the rPRL promoter fragments tested. Of note, a -127 deletion which was devoid of any basal promoter activity was also induced 10-fold by forskolin. The forskolin effect was abolished when GH4 rat pituitary cells were cotransfected with a plasmid encoding a protein kinase A inhibitor, indicating protein kinase A is involved in the activation mechanism. These data document that both positive and negative effectors influence basal rPRL promoter activity. Furthermore, the minimum sequences required for pituitary-specific rPRL promoter activity are altered by intracellular cAMP levels. Taken together, the data indicate that hormone-activated and cell-specific factors may interact to establish a particular setpoint for rPRL gene expression. PMID- 2547157 TI - Acute and chronic regulation of phosphoenolpyruvate carboxykinase mRNA by insulin and glucose. AB - Using the well differentiated rat hepatoma Fao we have studied the regulation of phosphoenolpyruvate carboxykinase (PEPCK) mRNA by insulin and glucose and compared these results to glucose production as estimated by glucose release into the medium. Fao cells possess an active gluconeogenic pathway and, when grown in glucose-free medium, release glucose for over 8 h. Addition of the cAMP analog, 8 (4-chlorophenyl-thio) cAMP (8-CTP-cAMP) or increasing the concentration of dihydroxyacetone and oxaloacetate results in an increase in glucose release which can be suppressed by insulin at concentrations between 1 and 100 nM. These effect of cAMP and insulin are associated with parallel changes in the level of mRNAPEPCK. Insulin treatment reduces mRNAPEPCK levels in these cells by 80%; this effect is transient reaching a maximum at 2-4 h. Addition of glucose to cells grown in glucose-free (G-) medium produces a decrease in mRNAPEPCK which is similar in magnitude and kinetics to that produced by insulin. Conversely, when cells grown in normal medium are placed in G- medium mRNAPEPCK levels triple over a period of 8 h, then return toward the basal value. Cells grown in G- medium or in G- medium plus 10nM insulin for 1 yr exhibit only slightly increased levels of mRNAPEPCK and respond to both 8-CTP-cAMP, and insulin, although the response to 8 CTP-cAMP is slightly blunted. These data indicate that glucose and insulin can play independent roles in regulation of PEPCK gene expression, and that these regulatory effects are usually transient. PMID- 2547159 TI - Human placental 17 beta-hydroxysteroid dehydrogenase is homologous to NodG protein of Rhizobium meliloti. AB - The amino acid sequence of human placental 17 beta-hydroxysteroid dehydrogenase (17 beta-OH-steroid dehydrogenase) was found to be similar to that of the NodG protein of Rhizobium meliloti. The computer-based comparison score is 11.5 SD higher than that obtained with 2500 comparisons of randomized sequences of these proteins. The probability of getting such a score by chance is 6 x 10(-31). 17 beta-OH-steroid dehydrogenase is also similar to Klebsiella aerogenes ribitol dehydrogenase and Escherichia coli glucitol-6-phosphate dehydrogenase. We propose that the steroid recognition site on 17 beta-OH-steroid dehydrogenase evolved from an ancestral recognition site for polyols such as ribitol and glucitol-6 phosphate. PMID- 2547158 TI - Distinct adenosine 3',5'-monophosphate and phorbol ester-responsive signal transduction pathways converge at the level of transcriptional activation by the interactions of DNA-binding proteins. AB - Evidence is presented that distinct cellular signal transduction pathways involving cAMP-dependent protein kinase-A and phorbol ester-stimulated protein kinase-C coordinately modulate gene transcription through common as well as distinct cis-acting elements and DNA-binding proteins. When transfected and expressed in HeLa and placental JEG-3 cells, fusion reporter plasmids that differ only by a single base deletion or addition to interconvert the octameric cAMP responsive element TGACGTCA (CRE) to form the heptameric phorbol ester-responsive element TGACTCA (TRE) are differentially regulated by cAMP and phorbol esters [12 O-tetradecanoyl phorbol-14-acetate (TPA)]. Transcription directed by the CRE is stimulated by cAMP and not TPA, although the basal expression mediated by this element in JEG-3 and HeLa cells is augmented by endogenous protein kinase-C activity. In contrast, TRE mediates transcriptional responses to both cAMP and TPA, and the two agents together give synergistic responses. Inhibition of cAMP dependent protein kinase-A by expression of a minigene encoding a peptide inhibitor of A-kinase abolishes the response of TRE to cAMP alone as well as the cAMP-induced component of the synergistic response to treatment with both TPA and cAMP. Desensitization of the protein kinase-C dependent pathway by prolonged exposure of cells to phorbol esters eliminates the TPA-induced transcription by TRE and inhibits the TPA-induced component of the synergistic response to both cAMP and TPA. Therefore, both protein kinases, A and C, are involved in transcriptional activation by the TRE; the function of either kinase alone results in a moderate level of activity, but the combined results of both functionally stimulated kinases are synergistically positive. Electrophoretic mobility shift assays using whole extracts of JEG-3 cells indicate that a common factor(s) binds both TRE and CRE; however, another factor(s) that binds to the CRE will not bind to the TRE. Further, a latent regulatory enhancer element (URE) located upstream of the CRE's in the human alpha gonadotropin gene, although inactive when paired alone with the alpha 100 promoter, induces basal and stimulated transcriptional activity of both CRE and TRE on the average of 10- to 20-fold. The data support the existence of a gene regulatory network consisting of related cis-acting elements and DNA-binding proteins whose transcriptional activities are regulated by the convergent actions of protein kinases-C and -A. PMID- 2547160 TI - Treatment of vaginal candidosis with fluconazole. AB - In patients with vulvovaginal mycosis systemic treatment is being discussed for eradication of possible fungal reservoirs, therefore, the efficacy and toleration of fluconazole, a triazole-derivative, has been investigated in the course of an open, comparative study. 104 patients with clinical, microscopic and cultural evidence of vaginal mycosis were included, 53 of whom were treated with a single dose of 150 mg of fluconazole and 51 patients daily received local treatment with 50 mg of econazole over a period of 6 days. The results regarding clinical efficacy were comparable for both preparations. At follow-up visits 4 to 10 d and 30 to 45 d post therapy the mycological as well as the subjective cure rates were similar. However, there was a higher mycological cure rate in the rectum of fluconazole patients which was statistically verifiable. This enables the elimination of rectal fungal infection as a source of homologous reinfection. PMID- 2547161 TI - Dioxin and health. Agent Orange study blasted. PMID- 2547162 TI - Ubiquitin and dementia. PMID- 2547163 TI - A novel genetic system to detect protein-protein interactions. AB - Protein-protein interactions between two proteins have generally been studied using biochemical techniques such as crosslinking, co-immunoprecipitation and co fractionation by chromatography. We have generated a novel genetic system to study these interactions by taking advantage of the properties of the GAL4 protein of the yeast Saccharomyces cerevisiae. This protein is a transcriptional activator required for the expression of genes encoding enzymes of galactose utilization. It consists of two separable and functionally essential domains: an N-terminal domain which binds to specific DNA sequences (UASG); and a C-terminal domain containing acidic regions, which is necessary to activate transcription. We have generated a system of two hybrid proteins containing parts of GAL4: the GAL4 DNA-binding domain fused to a protein 'X' and a GAL4 activating region fused to a protein 'Y'. If X and Y can form a protein-protein complex and reconstitute proximity of the GAL4 domains, transcription of a gene regulated by UASG occurs. We have tested this system using two yeast proteins that are known to interact- SNF1 and SNF4. High transcriptional activity is obtained only when both hybrids are present in a cell. This system may be applicable as a general method to identify proteins that interact with a known protein by the use of a simple galactose selection. PMID- 2547164 TI - Genetic analysis of immortalizing functions of Epstein-Barr virus in human B lymphocytes. AB - Epstein-Barr virus (EBV), a herpes virus, infects human B lymphocytes in vitro and efficiently immortalizes them. About 10 of the approximately 100 genes of EBV are expressed in recently immortalized B cells and although there is circumstantial evidence that at least three of these may contribute to the process of immortalization, there is no direct evidence that any particular gene is required. We have developed a genetic analysis of EBV that uses a transformation-defective strain of the virus as a helper virus in conjunction with DNA that contains all of the viral cis-acting elements required for replication, cleavage and packaging during the lytic phase of the viral life cycle. This DNA can include viral genes required for immortalization that complement the transformation-defective virus strain. The DNA can be amplified and packaged by the products of the helper virus and the packaged DNA is infectious. We have analysed two viral genes expressed in immortalized cells and find that the gene encoding EBV nuclear antigen-2 is required for immortalization, whereas the gene for the EBV nuclear antigen leader protein is not. PMID- 2547165 TI - Rapid increase of an immediate early gene messenger RNA in hippocampal neurons by synaptic NMDA receptor activation. AB - Recent studies in invertebrates indicate that a rapid genomic response to neuronal stimulation has a critical role in long-term changes in synaptic efficacy. Because several of the genes (immediately early genes; IEGs) that respond rapidly to growth factor stimulation of vertebrate cells in vitro are also activated by neuronal stimulation in vivo, attention has focused on the possibility that they play a part in synaptic plasticity in vertebrate nervous systems. Four IEGs thought to encode transcription factors, zif/268 (also termed Egr-1, NGFI-A, Krox 24), c-fos, c-jun, and jun-B are rapidly induced in the brain by seizure activity, and we have now studied the induction of these genes in a well-characterized model of synaptic plasticity in the vertebrate brain--long term potentiation (LTP) of the perforant pathgranule cell (pp-gc) synapse in vivo. We found that high-frequency (but not low-frequency) stimulation of the pp gc synapse markedly increases zif/268 messenger RNA (mRNA) levels in the ipsilateral granule cell neurons; mRNA of c-fos, c-jun and jun-B is less consistently increased. The stimulus frequency and intensity required to increase zif/268 mRNA levels are similar to those required to induce LTP, which is also seen only ipsilaterally, and both responses are blocked by NMDA-receptor antagonists as well as by convergent synaptic inhibitory inputs already known to block LTP. Accordingly, zif/268 mRNA levels and LTP seem to be regulated by similar synaptic mechanisms. PMID- 2547166 TI - [Influence of monoamines on heterosynaptic facilitation in giant neurons of Planorbis corneus]. AB - Monoamines (NE, DA) were studied for their effect on heterosynaptic facilitation of n-cholinergic synaptic transmission in identified giant neurons of the cerebral ganglion of Planorbis corneus. It was shown that NE and DA blocked n cholinergic EPSP and facilitated acetylcholine potential evoked by ionophoretic acetylcholine application. The monoamines blocked heterosynaptic facilitation as well. Washing of the monoamines after blocking heterosynaptic facilitation evoked recovery and long maintenance of the synaptic facilitation. It is concluded that DA and NE release may be the mechanism for conducting the intensity and duration of the heterosynaptic facilitation in the natural activity of the central nervous system of molluscs. PMID- 2547167 TI - [Restoration of potential-dependent anti-brain antibody-inhibited calcium current by cyclic adenosine monophosphate]. AB - A two-microelectrode voltage clamp method was used for measuring the voltage dependent calcium current (ICa) in isolated nonidentified snail neurons. Intracellular injection of cyclic adenosine monophosphate (cAMP, 10nA, 5 min) and extracellular application of dibutyryl-cAMP (dcAMP, I mmol/l, 10-20 min) did not change the normal ICa or reversibly decreased the ICa amplitude by 10-20%. Extracellular application of antibodies against S-100 proteins resulted in a Ca2+ dependent inactivation of the ICa: the ICa amplitude dropped to 15 +/- 12% of its initial level; cAMP and dcAMP restored an amplitude of the inhibited ICa up to 50 +/- 11%. It is shown that the effect of cAMP on ICa of intact neurons depends on the cytoplasmic Ca2+-level. PMID- 2547168 TI - [Action of the leukotriene LTC4 on the calcium channels of the somatic membrane of the nerve cell and tone of smooth muscle organs]. AB - Effects of synthetic leukotriene LTC4 on the internally perfused voltage-clamped snail neurons as well as on the electrical and mechanical activities of the stomach smooth muscle cells were studied. It was found that LTC4 increased calcium inward current in the soma membrane of a cell by activation of both voltage and receptor operated Ca-channels. LTC4 was effective only from outside but not from inside of the membrane which indicated that there was a receptor for leukotrienes in the neuronal membrane. PMID- 2547169 TI - [An attempt to estimate various characteristics of neuro-muscular transmission from the rising phase of miniature end-plate currents]. AB - A simple expression describing the relation between the duration of rising phase and amplitude of miniature end-plate currents (MEPC) is suggested. Parameters of MEPC of the rat diaphragm with different degrees of acetylcholinesterase inhibition by galanthamine were used to estimate the lower levels of acetylcholine diffussion coefficient (0.86.10-6 cm2/s) and channel opening rate constant (21000 s-1). PMID- 2547170 TI - Up-regulation of opiate receptors following chronic naloxone treatment in aged rats. AB - The present study assessed whether there are age-dependent differences in up regulation of opiate receptors following chronic naloxone treatment in mature (3 months) and aged (27 months) male Wistar rats. Half of each age group were implanted subdermally with slow-release naloxone pellets for 10 days, and half were given sham surgery. Twenty-four hours after pellet removal, the rats were decapitated and various CNS areas including spinal cord, hindbrain, midbrain, diencephalon, hippocampus, striatum, olfactory tubercles/nucleus accumbens and prefrontal cortex were dissected and assayed for [3H]naloxone binding. The results indicated that aged rats had fewer opiate receptors in the spinal cord, midbrain, striatum, and olfactory tubercle/nucleus accumbens. Despite this age related decline in opiate receptors, aged rats showed an up-regulation response similar to mature rats in all areas except the hippocampus of the left hemisphere, where they showed enhanced up-regulation relative to mature rats. The maintained plasticity of the opioid system contrasts with findings in other receptors systems where age-related impairments of antagonist-induced up regulation have been reported. PMID- 2547171 TI - Nerve growth factor receptor and choline acetyltransferase remain colocalized in the nucleus basalis (Ch4) of Alzheimer's patients. AB - Previous investigations have demonstrated an almost exclusive "coupling" between the receptor for nerve growth factor and cholinergic neurons within the basal forebrain. The present series of experiments were carried out to address two questions. First, what is the status of nerve growth factor receptor-containing neurons within the basal forebrain of patients with histopathologically confirmed diagnoses of Alzheimer's disease (AD)? More importantly, the second experiment assesses the degree to which the receptor for nerve growth factor and choline acetyltransferase remain colocalized within AD basal forebrain. A "decoupling" of this relationship, in which nerve growth factor receptors are no longer present upon magnocellular cholinergic neurons, would suggest that a loss of trophic support is functionally antecedent to the neuronal shrinkage and neuronal death seen in the basal forebrain in AD. Data obtained from six AD cases and four normal controls demonstrated an extensive reduction in number and shrinkage in size of nerve growth factor receptor containing neurons within the Ch4 region of the basal forebrain. Double label studies using either immunofluorescence or immunoperoxidase techniques demonstrated that the receptor for nerve growth factor and choline acetyltransferase remain colocalized in AD patients. This was true for neurons exhibiting either healthy or dystrophic morphological profiles. These data confirm previous studies, demonstrating that both a loss and shrinkage of cholinergic neurons occurs within the AD basal forebrain. The results of the present immunohistochemical investigation suggest that the degenerative changes associated with these neurons do not result from impaired trophic support related to a loss of NGF receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547173 TI - Functional change in the 5-HT presynaptic receptor in spinal cord of aged rats. AB - The present study assessed the functional integrity of the serotonin (5-HT) presynaptic receptor in the spinal cord of aged (18-20 months) rats. Previous research determined that exogenous 5-HT and 5-HT1B agonists, in adult (3 months) rats, activated the 5-HT presynaptic receptor resulting in a dose-dependent decrease in 3H-5-HT release from spinal cord slices. Contrastively, exogenous 5 HT and the selective 5-HT1B agonist 1-(m-chlorophenyl) piperazine HCl (mCPP) produced a dose-dependent increase in depolarization-evoked 3H-5-HT release from spinal cord of aged rats. The selective 5-HT1A agonist 8-hydroxy-2(di-n propylamino) tetralin (8-OHDPAT) increased 3H-5-HT release from aged rat spinal cord slices similar to results previously obtained from adult rat spinal cord slices. The inability of 5-HT and mCPP to inhibit 3H-5-HT release from spinal cord of aged rats indicates a functional change in the 5-HT presynaptic receptor in these animals. Possible mechanisms mediating this functional change are discussed. PMID- 2547172 TI - Localization of nerve growth factor receptors in the normal human brain and in Alzheimer's disease. AB - NGF receptors were visualized in human brain sections with an immunohistochemical procedure using a monoclonal antibody. This method results in the selective visualization of a population of neurons in the medial septal nucleus, the nucleus of the diagonal band of Broca, and the nucleus basalis of Meynert. Several lines of evidence indicate that this neuronal population is identical to the cholinergic neurons of the basal forebrain. NGF receptor immunohistochemistry therefore represents a sensitive and reliable procedure to selectively visualize forebrain cholinergic neurons for post-mortem analysis. NGF receptors were found to be expressed during the entire life span. However, the intracellular staining intensity was reduced in normal aging, suggesting the tentative conclusion that NGF receptor synthesis may decline in the aged brain. In Alzheimer's disease, the number of NGF receptor-positive cells was decreased. The morphological characteristics of surviving neurons were similar to immuno-positive neurons visualized in normal aged brains. PMID- 2547174 TI - Plasma adrenocorticotropic hormone in the rat demonstrates three different rhythms within 24 h. AB - Secretion of adrenocorticotropic hormone (ACTH) by the anterior pituitary is rhythmic and episodic, as reflected by fluctuations in plasma concentrations of ACTH. The present work was designed to further characterize the patterns of ACTH secretion that occur simultaneously within a 24-hour period in the rat. To accomplish this, blood collection protocols with sampling intervals of 2 min, 15 min, and 4 h were used in awake, chronically cannulated rats. Plasma samples were assayed for immunoreactive ACTH, and resultant data were analyzed for significant pulsatile secretory episodes. We observed three different patterns of ACTH secretion within a 24-hour period. Circadian variation occurred with peak plasma ACTH levels in the early evening. In addition, plasma ACTH exhibited two types of episodic variation: (1) episodic bursts with variable amplitudes that occurred approximately three times per hour which have been referred to as 'micropulses', and (2) prolonged elevations of plasma ACTH that occurred approximately 14 times in 24 h which have been referred to as 'larger ultradian' secretory episodes. These latter episodes appeared to consist of groups of relatively high amplitude micropulses. The physiological significance, functional interactions, and location of the controlling oscillator(s) of these different rhythms remain to be determined. PMID- 2547175 TI - Spontaneous spiking activities of porcine pars intermedia cells: effects of thyrotropin-releasing hormone. AB - The electrophysiological properties of porcine melanotrophs in primary culture were studied with patch clamp techniques. In the cell-attached (c/a) configuration, extracellular spikes were recorded but in only 47 of the 259 cells examined; active cells were more frequently found in short-term cultures (48 h, 22 out of 71). In the whole-cell (WCR) configuration, the mean resting potential was -46.6 mV and the mean input resistance 2.41 G omega. In the current clamp mode, of the 99 cells recorded, 39.4% displayed spontaneous spiking activities, 14.1% spiked when a depolarizing current was applied, and 46.5% were silent. Two patterns of spontaneous activity were recorded. The first consisted of rapid membrane depolarizations (mean duration: 16.0 ms) which reached a mean value of +8.9 mV. These action potentials occurred either at random intervals or at a mean frequency of 1.19 Hz. Tetrodotoxin (TTX), a sodium channel blocker, completely abolished these action potentials. The second pattern consisted of regular bursts (mean duration: 1.69 s and mean frequency: 0.15 Hz) of spikes with smaller amplitudes (culminating at -9.5 mV) and greater durations (79.7 ms). This pattern could be recorded in the presence of TTX in the external medium. In the c/a configuration, thyrotropin-releasing hormone (TRH) and (3Me-His2)-TRH enhanced spike frequency, whereas histidyl-proline-diketopiperazine, a metabolite of TRH, had no effect. In WCR, out of the 27 cells tested, TRH (5.10(-9)-5.10(-8) M) induced firing in 4 quiescent cells and increased the frequency of action potentials in 4 spontaneously active cells. This was usually, but not necessarily, preceded by a hyperpolarization (n = 5). TRH (5.10(-8) M) enhanced the secretion of melanocyte-stimulating hormone (alpha MSH) from perfused isolated melanotrophs by 62.8 +/- 9.8% (n = 9) over basal levels. (3Me-His2)-TRH and (Pro3)-TRH mimicked the TRH response, whereas histidyl-proline diketopiperazine was without effect. PMID- 2547176 TI - Estrogen modulation of the alpha-1-adrenergic response of hypothalamic neurons. AB - Intracellular recordings were made from 106 arcuate and cell-poor zone (ARC-CPZ) neurons in sagittal slices prepared from intact, ovariectomized and ovariectomized plus estradiol-benzoate-treated female guinea pigs, and the effects of norepinephrine (NE), the alpha 1-agonist methoxamine (MX) and the beta agonist isoproterenol were tested. Either bath application or pressure application of 2-100 microM NE reversibly hyperpolarized and inhibited the spontaneous firing of the majority (57%, n = 60) of ARC-CPZ neurons. Isoproterenol also inhibited the majority (75%) of the ARC-CPZ neurons which it was tested on. In addition, 2-100 microM NE depolarized and/or increased the spontaneous activity of 20% (n = 21) of ARC-CPZ neurons, and some of these (n = 8) exhibited bursting activity. Similar doses of MX mimicked the NE excitation (depolarization and/or increased firing) in 48% (n = 14) of the ARC-CPZ neurons tested. Based on the serum levels of 17 beta-estradiol, the three groups of females were divided into high ( greater than 30 pg/ml) and low (less than 30 pg/ml) estrogen groups, and it was found that endogenous or exogenous estrogen significantly increased the number of neurons responding to MX (from 29 to 75%). Using intracellular labeling with procion yellow and immunocytochemistry, we have identified that luteinizing-hormone-releasing hormone neurons respond to NE. Therefore, it is suggested that one mechanism for an increase in the noradrenergic excitatory drive at the time of the preovulatory surge of luteinizing hormone in the mammal is an increase in the neuronal response to alpha 1-adrenergic stimulation. PMID- 2547177 TI - Evidence for prolonged suppression of stress-induced release of adrenocorticotropic hormone and corticosterone with a brain-enhanced dexamethasone-redox delivery system. AB - We have developed a redox system for brain-enhanced delivery of dexamethasone based on an interconvertible dihydropyridine in equilibrium pyridinium salt carrier. Dexamethasone, when combined with the lipoidal carrier, readily crosses the blood-brain barrier. The carrier, when oxidized, reduces its rate of exit from the brain. The aim of the study was to evaluate the capacity of a dexamethasone-chemical delivery system (DX-CDS) and dexamethasone (DEX) to suppress stress-induced elevations of plasma adrenocorticotropic hormone (ACTH) and corticosterone (CORT). Adult male Sprague-Dawley (CD) rats were administered either DX-CDS (10 mg/kg), an equimolar dose of DEX or the drug vehicle (2 hydroxypropyl-beta-cyclodextrin) by a single tail vein injection. Rats then received either no stress or a restraint stress for a 5- or 15-min duration on days 1, 3, 5 or 7 after drug administration and trunk blood was rapidly collected. To assess peripheral effects of DX-CDS and DEX, 1 ml of blood was removed via orbital puncture and evaluated for total and differential leukocyte counts in a separate group of animals. Both DX-CDS and DEX were effective on day 1 in suppressing, by greater than 95%, ACTH secretion induced by a 5-min stress. However, DX-CDS was effective through day 5 (44% suppression) while DEX was not effective after 24 h. When 15 min of stress was applied, DX-CDS effected a significant ACTH suppression through 7 days while DEX was effective for only 3 days. DX-CDS was effective through day 7 (55%) in suppressing CORT after a 15-min stress while DEX was effective for 3 days only.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547179 TI - Protection of cell proteins against free-radical attack by nootropic drugs: scavenger effect of pyritinol confirmed by electron spin resonance spectroscopy. AB - The potency of nootropic drugs to protect cell proteins and lipids against free radical attack was studied. In an in vitro system, generating hydroxyl radicals by a Fenton-type reaction, the soluble proteins (bovine serum albumin and cytosol protein from brain) were quickly insolubilized and precipitated. Pyritinol (and tamitinol) exhibited the best protection against insolubilization of protein while centrophenoxine (meclophenoxate) and its dimethylaminoethanol moiety were less effective, piracetam (and oxiracetam) being without effect. The lipid peroxidation induced by free radicals from cyclic redox reactions of iron ascorbate was not influenced by pyritinol, indicating the selectivity of its scavenger action. The efficient scavenging of hydroxyl radicals by pyritinol was confirmed by electron spin resonance spectroscopy measurement of hydroxyl radicals entrapped by spin trap. Millimolar concentrations of pyritinol competitively decreased the formation of spin adducts. The results suggest that the protective effect of pyritinol against free-radical induced derangement of cell proteins may be an important part of its antirheumatic, as well as nootropic, action. PMID- 2547178 TI - Ovarian nerve extracts influence androgen production by cultured ovarian thecal cells. AB - There is growing evidence that ovarian steroidogenesis is controlled not only by pituitary gonadotropins but also by ovarian nerves. Nerves reach the ovary via the plexus nerve and via the superior ovarian nerve (SON), which runs in the suspensory ligament, and innervate theca cells of all sizes of follicles. To investigate the role of ovarian nerves in steroidogenesis we have examined the effects of adding an extract of SON from adult rats on androgen production by cultured porcine theca cells. Addition of SON extract to cultured theca interna from 3 to 6 mm diameter follicles of prepubertal gilts significantly inhibited (p less than 0.05) LH-stimulated androstenedione production in a dose-dependent manner; significant inhibition (10.8%) occurred with the addition of the extract of 2 mg of SON/ml culture medium, and near maximal inhibition (83%) resulted when the SON extract was increased to 60 mg SON/ml. Extracts of sciatic nerves, used as non-ovarian nerve controls, failed to inhibit, and in fact significantly increased (p less than 0.05) androstenedione production over the same concentration range of neural tissue extract. The inhibitory effect of the SON extract was unaffected by chymotrypsin digestion or by the presence of the beta adrenergic antagonist propranolol (10(-6) M), but was removed by charcoal treatment. These results suggest that the nervous system has the potential for modulation of follicular steroid biosynthesis via direct innervation of the ovaries, in addition to the well-established indirect mechanism of neural control exerted via the hypothalamic-pituitary system. PMID- 2547180 TI - The effects of tetraethylammonium during twitch and tetanic stimulation of the phrenic nerve diaphragm preparation in the rat. AB - Tetraethylammonium (TEA) (2.6 x 10(-3) M) potentiated the twitches of the indirectly- or directly-stimulated phrenic nerve diaphragm of the rat at 37 degrees C by prolonging the action potential of the sarcolemma, due to an inhibition of the repolarizing K+ current. With indirect stimulation, TEA caused a use-dependent inhibition of tetanic contractions, induced every 10 min by 10 sec of 50 Hz stimulation, and a post-tetanic depression of the twitches was observed after about 40 min. Recording of the electromyogram (EMG) and compound action potentials of the phrenic nerve, localized the two inhibitory effects to the neuromuscular junction. They were caused by different mechanisms of action. Choline (3.6 x 10(-4) M) antagonized the depression of the twitch but not the use dependent inhibition. Lowering the temperature to 20 degrees C reduced the depression of the twitch, whereas the use-dependent inhibition was enhanced. The release of transmitter was probably normal during tetanic stimulation; a post synaptic desensitization of acetylcholine (ACh) receptors caused the inhibition. Microelectrode recordings of endplate potentials supported this conclusion. The depression of the twitch was due to a presynaptic depletion of transmitter. This was confirmed by inducing an additional depletion and depression of the twitch with N-ethyl-maleimide (2.5 x 10(-5) M). Since the depression of the twitch was antagonized by choline, the depletion was probably due to an inhibited uptake of choline into the nerve terminals. PMID- 2547182 TI - Anterior cervical discectomy with hydroxylapatite fusion. AB - The outcome of microscopic anterior cervical discectomy with iliac crest interbody fusion in a group of 75 patients was compared with that of microscopic anterior cervical discectomy with synthetic hydroxylapatite fusion in a group of 84 patients. The rate of relief of myelopathy (70%) was similar in both groups, but those who underwent synthetic fusion had better long-term relief of radiculopathy, less need for a second operation at the same or an adjacent level, no resorption of the bone plug, comparable spinal alignment and stability, and the elimination of complications at the iliac crest donor site. The data suggest that hydroxylapatite fusion may be equal or superior to autologous iliac crest interbody fusion for anterior cervical disc surgery. PMID- 2547181 TI - The dopamine agonist, PHNO ((+)-4-propyl-9-hydroxynaphthoxazine), inhibits cyclic adenosine 3',5'-monophosphate formation and prolactin release from anterior pituitary cells. AB - In cultured rat anterior pituitary cells incubated for 30-180 min, PHNO caused a concentration-dependent inhibition of prolactin release (IC50 approximately 0.5 nM) with maximal suppression at 5 nM. Forskolin increased cyclic adenosine 3',5' monophosphate (cAMP) generation by stimulating adenylate cyclase and PHNO inhibited this effect with the same concentration profile as for inhibition of prolactin release. Inhibitory effects of 0.5 nM PHNO on prolactin release and cAMP generation were abolished by coincubation with 10 nM haloperidol, a D2 dopamine receptor antagonist. Within 30 min, 0.5 nM PHNO blunted the stimulation of prolactin release due to 10 nM thyrotropin-releasing hormone (TRH) or angiotensin II (AII). Thus, PHNO appears to activate the D2 dopamine receptor to inhibit the formation of cAMP and the secretion of prolactin. PMID- 2547183 TI - Epithelial dysplasia produced by carcinogen pretreatment and subsequent wounding. AB - Golden hamsters in which the tongues were pretreated with DMBA for 4 weeks, subsequently excised, and either received no treatment or received additional postexcisional applications of acetone or DMBA for 9 to 13 days exhibited histologic evidence of epithelial dysplasia; these dysplastic lesions stained positive for papillomavirus genus-specific antigens. Conversely, animals in which the tongues received the same pretreatment yet had no excision failed to show any dysplastic changes, even though the tongues received additional applications of DMBA. Furthermore, papillomavirus genus-specific antigens were not demonstrable. The animals in which the tongues received no pretreatment with DMBA for 4 weeks followed by excisional wounding showed normal healing, even though the tongues were post-treated with DMBA. Papillomavirus genus-specific antigens could not be demonstrated in this last group. PMID- 2547184 TI - c-fos expression induces bone tumors in transgenic mice. AB - The proto-oncogene c-fos has been isolated as the cellular homolog of the v-fos gene found in the osteosarcoma inducing FBR- and FBJ-murine sarcoma viruses (MSV). Expression of the c-fos gene in transgenic mice leads to the development of bone lesions of which about half progress to bone tumors mainly chondrosarcomas. The tumors display a strong preference for males and have a latency with a mean of 9.5 months. However, also mice without visible lesions develop bone tumors with the same sex preference and latency. These consequences of c-fos expression are independent of the chosen promoter but dependent on a replacement of 3' noncoding sequences of c-fos by a long terminal repeat (LTR) of the FBJ-MSV virus. PMID- 2547185 TI - Primary placement of osseointegrated implants in microvascular mandibular reconstruction. AB - The goal of mandibular reconstruction is to rehabilitate the patient by restoring occlusal relationships, lower facial contour, oral continence, and a denture bearing surface. One of the major advantages of the use of vascularized bone over all other methods of mandibular reconstruction is its ability to achieve dental rehabilitation rapidly. The use of osseointegrated dental implants is a valuable adjunct in oral rehabilitation. It provides the most rigid form of stabilization to withstand the forces of mastication. In situations in which soft tissue reconstruction or the height of the alveolar ridge is not sufficient for a tissue borne denture, implants offer the most suitable alternative. Mandibular reconstruction with free tissue transfer techniques is ideally suited for the placement of implants. These can be inserted at the time of mandibular reconstruction. Four months after surgery, when the integration process has occurred, the implants are unroofed, loaded, and ready for prosthetic placement. We will present several representative patients who underwent mandibular reconstruction with microvascular free bone transfer who have been successfully rehabilitated by osseointegrated implants. The process of osseointegration, different types of dental implants, and issues regarding radiation and implants are discussed. This is the first report of dental rehabilitation by primary placement of dental implants in patients undergoing microvascular mandibular reconstruction. PMID- 2547187 TI - Angioedema of the head and neck and angiotensin-converting enzyme inhibitors. PMID- 2547186 TI - Cerebral shift after lateral craniofacial resection and flap reconstruction. AB - The biomechanical effects on the cranial contents after craniectomy and immediate soft-tissue flap reconstruction are substantial and potentially life threatening. Once the rigid protective covering of the skull is removed, the cerebral component of the cranial contents is vulnerable to rising extrinsic pressure. Intracranial pressure falls, brain compliance rises, and contralateral cerebral shift may occur by virtue of the craniectomy alone. However, if flaps filling the defect should compress by weight or swelling, even greater midline or craniocaudal shift of the brain may occur. We present three cases in which midline cerebral shift was documented by CT scan after flap reconstruction of lateral craniofacial/craniotemporal resection. Three other patients undergoing anterior craniofacial resection during the same time period for skin cancer (one patient) and esthesioneuroblastoma (two patients) had no shift in intracerebral contents. Symptomatic or asymptomatic intracranial shift may occur soon after substantial craniectomy and soft-tissue flap reconstruction. Midline shift from lateral resection and reconstruction is more likely than anteroposterior shift from anterior craniofacial resection. Strategies for minimizing this compression or shift are described. PMID- 2547188 TI - Oropharyngeal angioedema associated with the use of angiotensin-converting enzyme inhibitors. AB - Angioedema that occurs in the head and neck may be a serious and possibly life threatening condition. We present two cases of oropharyngeal angioedema associated with the use of angiotensin-converting enzyme inhibitors in patients with no underlying renal failure. Angiotensin-converting enzyme-inhibiting agents are now commonly used to control hypertension. Treatment of acquired angioedema is directed first at protecting the airway by careful observation, intubation, or if necessary, tracheostomy. The causative agent is withdrawn, and the patient is treated with antihistamines and steroids until the acute episode resolves. Although not used in our patients, the treatment of angioedema with subcutaneous epinephrine has been described. As more patients are treated with angiotensin converting enzyme inhibitors, it becomes essential for the otolaryngologist to become aware of this potentially life-threatening complication. PMID- 2547189 TI - Attempts to gauge the relative importance of pre- and postsynaptic effects of morphine on the transmission of noxious messages in the dorsal horn of the rat spinal cord. AB - Both pre- and postsynaptic mechanisms have been proposed as an explanation of the depressive effects of opioids on the activity of nociceptive dorsal horn neurons. In order to gauge the importance of the two mechanisms, we studied the effect of morphine on the spontaneous hyperactivity of nociceptive dorsal horn neurons in the spinalized decerebrated deafferented rat (C5-Th1). In this preparation, intravenous morphine was shown to depress spontaneous firing rate in a dose dependent fashion. A comparative analysis of the effect of the same dose of morphine (2 mg/kg i.v.) in the intact spinalized decerebrated arthritic rat, in which dorsal horn convergent neurons also display high spontaneous activity, revealed that systemic morphine is twice as potent when primary afferent fibers are left intact. These results can explain why the analgesic effect of morphine is more marked against pains due to an excess of nociception than against pains arising from deafferentation. PMID- 2547190 TI - Metastatic Wilms' tumor to the liver with MR findings simulating cysts: case report emphasizing need for integrated imaging. AB - A case of Wilms' tumor metastatic to the liver with previously undescribed MR findings is presented. Although there is no evidence that there were hepatic metastases at presentation in this case, small lesions with this appearance would be difficult to detect with MRI. This could lead to incorrect staging of Wilms' tumor examined by MRI alone. PMID- 2547191 TI - Ganciclovir treatment of cytomegalovirus disease in immunocompromised children. AB - Twelve immunocompromised children were treated with 13 courses of intravenously administered ganciclovir for severe cytomegalovirus disease. All children were allograft recipients; 6 received organ transplants (5 liver, 1 kidney) and 6 received bone marrow. They presented with one or more of the following forms of cytomegalovirus disease: pneumonitis, 9; hepatitis, 3; colitis, 2; peritonitis, 1; and retinitis, 1. Clinical improvement was observed in 7 (58%) of 12 patients during ganciclovir therapy. Cessation of active viral replication during therapy accompanied 69% of the treatment courses. Mild and transient increases in creatinine and liver function tests and/or decreases in neutrophil count accompanied 77% of treatment courses but neutropenia (less than 1000 cells/mm3) did not occur. Transient decreases in platelet counts accompanied therapy in 3 bone marrow allograft recipients, but greater than 50% decrease in lymphocyte count was not seen. We conclude that ganciclovir is safe and appears to have a beneficial effect on cytomegalovirus disease in some pediatric transplant recipients. PMID- 2547192 TI - Prenatal diagnosis of fetal infection: advances from amniocentesis to cordocentesis--congenital toxoplasmosis, rubella, cytomegalovirus, varicella virus, parvovirus and human immunodeficiency virus. PMID- 2547193 TI - Okadaic acid, a phosphatase inhibitor, produces a Ca2+ and calmodulin-independent contraction of smooth muscle. AB - The effects of okadaic acid, a phosphoprotein phosphatase inhibitor, on the contractile response and on myosin light chain phosphorylation were studied in intact lamb tracheal smooth muscle. The effects of okadaic acid were compared to the response of the same fibers stimulated with 1 microM methacholine, a concentration that induces 90% of maximal force. Okadaic acid (50 microM) produced a slow but maximal contraction that was accompanied by an increase in phosphorylation of the 20 kDa light chain of myosin. The myosin light chain phosphorylation pattern induced by okadaic acid, however, differed from that induced by methacholine. Ca2+ depletion, N-(6-aminohexyl)-5-chloro-1 naphthalenesulfonamide (W-7), a calmodulin antagonist and 1-(5 isoquinolinylsulfonyl)-2-methylpiperazine (H-7), a protein kinase C inhibitor, blocked or attenuated methacholine-induced contractions but had no significant effect on force development or myosin light chain phosphorylation induced by okadaic acid. These results suggest that phosphorylation of the 20 kDa light chain of myosin is essential for smooth muscle contraction; they also suggest that okadaic acid either uncovers or activates an apparently Ca2+ and calmodulin independent protein kinase activity that phosphorylates the 20 kDa light chain of myosin at multiple sites. PMID- 2547194 TI - Modulation of Ca-channel current by an adenosine analog mediated by a GTP-binding protein in chick sensory neurons. AB - Inhibitory modulation of the high-voltage-activated (HVA) Ca-channel current by 2 chloroadenosine (2CA) was studied in chick sensory neurons using the whole-cell clamp method. 2CA reduced the omega CTX-sensitive HVA-current (Aosaki and Kasai 1989) in a dose-dependent manner with a Kd of 0.8 microM. The inhibition by 2CA was also voltage-dependent, being maximal at hyperpolarized potentials, and completely removed at potentials more positive than 30 mV. This voltage dependence of 2CA action was also evident as a progressive increase in Ca-channel current magnitude during a depolarization which could be described by a single exponential function and which became faster at larger depolarizations. The concentration of 2CA affected the steady-state reduction in Ca-channel current, but did not alter the time-course of current increase during depolarization. The voltage-dependent effect of 2CA was mimicked by intracellular application of GTP gamma S, but not by phorbol ester, arachidonic acid or nitroprusside. These results are consistent with model in which 2CA activates a G-protein, which then unmasks an additional activation gate on the Ca-channel. PMID- 2547195 TI - Characterization of two kinds of high-voltage-activated Ca-channel currents in chick sensory neurons. Differential sensitivity to dihydropyridines and omega conotoxin GVIA. AB - High-voltage-activated (HVA) Ca-channel currents in chick sensory neurons were characterized by dihydropyridine compounds (DHPs) and omega-conotoxin GVIA (omega CTX) using patch-clamp methods. In single-channel recordings, two HVA-currents were identified by their single-channel conductances, 13 pS and 25 pS in 110 mM BaCl2. DHPs selectively affected the large-conductance channel. omega CTX (5 microM), on the other hand, irreversibly eliminated only the small-conductance channel, while the large-conductance channel was either unaffected or only transiently blocked. In whole-cell recordings the macroscopic HVA-current was completely and irreversibly blocked by omega CTX but insensitive to DHPs in 60% of the cells. This current presumably was carried by the 13 pS channel. In the remaining cells, a part of the HVA-current (10%, SD = 11%) was either unaffected or transiently blocked by omega CTX and was sensitive to DHPs. This current presumably was carried by the 25 pS channel. Inactivation of both macroscopic current component was incomplete during a 150 ms long depolarization. Our data suggest that the HVA-currents in chick sensory neurons are carried by two distinct Ca-channels that are differentially affected by omega CTX and DHPs. PMID- 2547196 TI - [Hepatocellular carcinoma associated with schistosomiasis japonica; CT and angiographic features]. AB - One-hundred and fourteen cases of hepatocellular carcinoma (HCC) which underwent computed tomography (CT) and angiography at the University Hospital of Yamanashi were reviewed. These included 24 cases of HCC associated with hepatic schistosomiasis japonica. Compared with the other cases, radiological features of HCC with schistosomiasis japonica were clarified. Patients with multinodular HCC were most common (13/24), followed by those with single nodular HCC (7/24), while those with massive or diffuse HCC were few (4/24). In patients with marked CT changes (grade III or IV) of hepatic schistosomiasis japonica, most of HCC nodules were disclosed as a mass with homogenous hypodensity surrounded by "shell like calcifications". Obstruction of the portal vein or its branches due to tumor thrombus was rarely noted (4/24). These features are presumably attributed to periportal fibrosis which is characteristic of heptic schistosomiasis japonica. Because of these features, it is expected that hepatocellular carcinomas with schistosomiasis japonica can be treated by transcatheter arterial embolization more effectively than those without schistosomiasis japonica. PMID- 2547197 TI - [Usefulness of bone scintigraphy on diagnosis of malignant fibrous histiocytoma of bone]. AB - We examined the roentgenographical features of 5 patients who were diagnosed histopathologically as malignant fibrous histiocytoma (MFH) of bone, especially on the imaging of bone scintigraphy. Two patients had multiple lesions on the initial examination, and two of 3 patients who showed solitary lesion on the first examination could be detected more pathological sites by bone scintigraphy which was studied within two months of the first investigation. On bone scintigraphy, the mixture of hot and cold lesions which was coincident with the osteolytic change on plan X-P could be noticed, and with the progression of osteolysis it had a tendency to increase the area of cold lesions. These features are similar to metastatic bone tumor which is the most questionable disease on diagnosis, and we conclude that bone scintigraphy will be absolutely necessary when MFH of bone is highly suspected in the clinical course. PMID- 2547198 TI - Treating minor burns. PMID- 2547199 TI - Lisinopril: a new angiotensin-converting enzyme inhibitor. AB - Lisinopril is a synthetic, nonsulfhydryl, angiotensin-converting enzyme inhibitor. Its bioavailability is approximately 25% and is not affected by food. Hepatic metabolism is not required for pharmacologic effect, which occurs 1 hour after administration. Peak serum concentration and effect are delayed, occurring 6-8 hours after a single dose and lasting for at least 24 hours. The drug is eliminated primarily by the kidneys. The elimination half-life is 12.6 hours and is prolonged in renal impairment. Lisinopril 10-80 mg once a day is effective in lowering blood pressure in all grades of essential and renovascular hypertension. It is as effective as hydrochlorothiazide, atenolol, metoprolol, and nifedipine. Combining lisinopril with hydrochlorothiazide produces a greater degree of blood pressure reduction. Patients with congestive heart failure have demonstrated immediate and prolonged beneficial hemodynamic effects and increased exercise tolerance. Lisinopril is well tolerated. Clinically significant drug interactions have not been reported, but caution should be used when lisinopril is administered with diuretics, nifedipine, or agents that may increase concentrations of potassium. The usual initial oral dosage of lisinopril is 10 mg once a day (range 20-40 mg/day). Lower dosages may be necessary in patients with renal impairment or congestive heart failure, elderly persons, and those receiving diuretics. PMID- 2547200 TI - Saturable mechanism for delta sleep-inducing peptide (DSIP) at the blood-brain barrier of the vascularly perfused guinea pig brain. AB - Cellular uptake of [125I] labelled DSIP at the luminal interface of the blood brain barrier (BBB) was studied in the ipsilateral perfused in situ guinea pig forebrain. Regional unidirectional transfer constants (Kin) calculated from the multiple-time brain uptake analysis were 0.93, 1.33 and 1.66 microliter.min-1 g-1 for the parietal cortex, caudate nucleus and hippocampus, respectively. In the presence of 7 microM unlabelled DSIP the brain uptake of [125I]-DSIP (0.3 nM) was inhibited, the values of Kin being reduced to 0.23-0.38 microliter.min-1 g-1, values that were comparable with the Kin for mannitol. The rapidly equilibrating space of brain, measured from the intercept of the line describing brain uptake versus time on the brain uptake ordinate, Vi, was greater for [125I]-DSIP than for mannitol; in the presence of unlabelled DSIP this was reduced to that of mannitol, and it was suggested that the larger volume for [125I]-DSIP represented binding at specific sites on the brain capillary membrane. L-tryptophan, the N terminal residue of DSIP, in concentrations of 7 microM and 1 mM, inhibited Kin without affecting Vi. A moderate inhibition of Kin was obtained by vasopressin ([Arg8]-VP), but only at a concentration as high as 0.2 mM. The results suggest the presence of a high affinity saturable mechanism for transport of DSIP across the blood-brain barrier, with subsequent uptake at brain sites that are highly sensitive to L-tryptophan, and may be modulated by [Arg8]-VP. PMID- 2547201 TI - One receptor type mediates two independent effects of FMRFa on neurosecretory cells of Lymnaea. AB - Structure activity relations (SAR) of FMRFa on the transient hyperpolarizing response and long lasting depression of excitability of neurosecretory caudo dorsal cells (CDCs) of the pond snail Lymnaea stagnalis were examined. Although these effects to FMRFa occur independently, the SARs for the induction of both responses were identical suggesting that CDCs possess a single type of FMRFa receptors. Native GDPFLRFa and SDPFLRFa were equipotent to FMRFa receptors. It is concluded that activation of the receptor requires [Arg3-Phe4]-NH2, whereas N terminal amino acids are involved in binding. PMID- 2547202 TI - Effects of two ACTH analogs on successive odor discrimination learning in rats. AB - Olfactory discrimination learning has been shown to provide a powerful tool to investigate the mechanisms involved in the formation, storage and retrieval of information in rodent CNS. In the present study we tested the effects of two ACTH analogs, which were previously reported to influence the processes of learning and memory, on various olfactory learning tasks. The ACTH(4-9) analog HOE 427 produced an apparent increase in storage of olfactory information as shown by the difficulty experienced by the animals to rapidly reverse their behavioral responses to previously learned odors. Similarly, the ACTH (4-9) analog ORG 2766 appears to enhance the storage of olfactory information when administered either before or after the learning trials. These data are consistent with the notion that ACTH and related analogs facilitate performance in a variety of learning tasks. In addition, our results suggest possible mechanisms by which some neuroactive peptides might modulate learning and memory processes in the CNS. PMID- 2547203 TI - [D-Ala2,Leu5,Cys6]enkephalin: short-term agonist effects and long-term antagonism at delta opioid receptors. AB - Four experiments were conducted. Two examined the in vivo short-term effects (less than 35 min) and two examined the in vivo long-term effects (greater than 2 days) of the synthetic enkephalin analogue, [D-Ala2,Leu5,Cys6]enkephalin (DALCE). In the short term, DALCE was found to produce analgesia and transient immobility after intracerebroventricular (ICV) administration. We found a dose-related increase in paw-lick latency for rats placed on a hotplate (52 degrees C). In the second experiment, we determined that immobility was attenuated by pretreatment with naltrexone methobromide (QNTX, 0.1 microgram), and the delta selective antagonist, 16-methyl cyprenorphine (M80, 5 micrograms). QNTX and M80 significantly attenuated DALCE-induced immobility by more than 50% of control. Paw-lick latency was then measured on the hotplate to assess analgesia. Pretreatment with M80 reliably attenuated the DALCE-induced analgesia. Whereas QNTX failed to reliably attenuate paw-lick latency on the first trial, it was as effective as M80 on the second trial. We suggest that the short-term agonist effects of DALCE are produced by actions at mu and delta opioid receptors as would be predicted from prior in vitro studies showing moderate to high affinity, respectively, at these receptors. In the third experiment, DALCE was shown to display long-term behavioral antagonism that was selective for the delta receptor. Rats were injected ICV with 6.7 micrograms of DALCE and tested 48 hr later. Analgesia was measured by injecting 15% formalin subcutaneously followed 20 min later by an ICV injection of one of three selective opioid agonists (DAGO, DPDPE or U50488H).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547204 TI - The effects of structure-conformation modifications of melanotropin analogs on learning and memory: D-amino acid substituted linear and cyclic analogs. AB - Alpha-MSH has a wide variety of putative biological activities in addition to its classical melanocyte dispersing activity. Since each of these activities appears to be mediated by a discrete receptor, this peptide is an excellent candidate for exploring conformational restrictions which determine the chemical-physical basis for hormone action on specific activities. Experiments One and Two evaluated several cyclic and linear analogs of alpha-MSH on retrieval of memory during the reactivation of memory for a passive avoidance response following hypothermia induced amnesia. Three of the cyclic analogs appear to have enhanced the peptide's ability to serve as a reactivation agent. One of the linear Nle4,D-Phe7 analogs antagonized whereas three others enhanced reactivation. The D-Phe7 substitution in cyclic analogs did not affect reactivation. Another group of animals were trained on a step-through passive avoidance task and tested 25 days later. The cyclic analog enhanced memory whereas the D-Phe7 analog and alpha-MSH had no effect. Finally, two analogs were tested on a black-white discrimination. Although the cyclic analog had no effect on either acquisition or reversal of this learning, the Nle4,D-Phe7 analog significantly impaired reversal learning. The results from these preliminary studies suggest that structural modifications of alpha-MSH do alter its potency and pattern of actions in learning and memory situations. PMID- 2547205 TI - Melanin-concentrating hormone (MCH) immunoreactivity in the brain and pituitary of the dogfish Scyliorhinus canicula. Colocalization with alpha-melanocyte stimulating hormone (alpha-MSH) in hypothalamic neurons. AB - The distribution of melanin-concentrating hormone (MCH) in the central nervous system of the dogfish Scyliorhinus canicula was determined by indirect immunofluorescence and peroxidase-anti-peroxidase techniques, using an antiserum raised against synthetic salmon MCH. Three groups of MCH-positive cell bodies were localized in the posterior hypothalamus. The most prominent cell group was detected in the nucleus sacci vasculosi. Scattered MCH-immunoreactive cells were observed in the nucleus tuberculi posterioris and in the nucleus lateralis tuberis. At the pituitary level, the caudal part of the median lobe of the pars distalis contained strongly MCH-positive perikarya. Some of these cells were liquor-contacting-type. Immunoreactive fibers originating from the hypothalamic perikarya projected throughout the dorsal wall of the posterior hypothalamus. Positive fibers were also detected within the thalamus and the central gray of the mesencephalon. The distribution of MCH-containing neurons was compared to that of alpha-MSH-immunoreactive elements using consecutive, 5-micron thick sections. Both MCH- and alpha-MSH-immunoreactive peptides were found in the same neurons of the nucleus sacci vasculosi. These data suggest that MCH and alpha MSH, two neuropeptides which exert antagonistic activities on skin melanophores, may also act in a coordinate manner in the central nervous system of cartilaginous fish. PMID- 2547206 TI - Functional corticotropin-releasing factor (CRF) receptors in mouse spleen: evidence from adenylate cyclase studies. AB - Radioligand binding studies have previously identified a high affinity, magnesium dependent, guanine nucleotide-sensitive binding site for corticotropin-releasing factor (CRF) in mouse spleen. In order to determine the functional nature of these CRF binding sites, we examined the effects of CRF on adenylate cyclase activity in mouse spleen homogenates. The stimulation of adenylate cyclase activity was dependent on time, tissue protein concentration, and guanine nucleotides. CRF-stimulated adenylate cyclase activity was evident in the presence of guanosine-5'-triphosphate (GTP) and its precursor guanosine-5' diphosphate (GDP) but was not detected in the presence of the hydrolysis resistant GTP analogs, guanyl-5'-imidodiphosphate [Gpp(NH)p] and guanosine-5' gamma-thiotriphosphate (GTP-gamma-S). The rank order of potency for CRF analogs and fragments in stimulating adenylate cyclase activity was comparable to their affinities for CRF binding sites in mouse spleen homogenates. The putative receptor antagonist, alpha helical ovine CRF(9-41), did not stimulate adenylate cyclase activity but did attenuate the stimulation by various concentrations of rat/human CRF. In summary, these data demonstrate the functional nature of CRF receptors in mouse spleen as evidenced by CRF stimulation of cAMP production and suggest that this peptide may play a physiological role in regulating immune function. PMID- 2547207 TI - Angiotensin converting enzyme: substrate inhibition. AB - Phosphate, borate, and Tris inhibit angiotensin converting enzyme (ACE), but HEPES buffer is inert. Measurements of substrate inhibition were made in HEPES buffer at pH 7.0 and 25 degrees C and 37 degrees C. Substrate inhibition was marked and goes to completion. A new equation for substrate inhibitions enables one, under favorable circumstances, to determine whether there is cooperativity in the binding of substrate to the inhibitory and active sites. Cooperativity does occur with ACE using Hipp-His-Leu as substrate. The kinetic parameters were measured (Km = 0.21 mM, K* = 0.65 mM at 37 degrees C). The enzyme concentration (1.94 X 10(-8) M) was determined by titration with lisinopril so that kcat (5 X 10(3) at 37 degrees C) could be determined. Using this value and the molecular weight the specific activity of ACE was calculated for different common buffers. The specific activity in HEPES calculated from Vmax was 33.7 units/mg at 37 degrees C. PMID- 2547208 TI - Comparison of neurotensin levels, receptors and actions in LS/Ibg and SS/Ibg mice. AB - Neurotensin (NT), injected centrally, markedly enhances sensitivity to ethanol induced anesthesia in SS but not in LS mice (4). Since LS and SS mice were bred selectively for differential sensitivity to ethanol, these findings suggest that neurotensinergic neuronal processes mediate some of ethanol's actions and that LS and SS mice might differ genetically in neurotensinergic systems. Indeed, in biochemical studies it was shown that LS and SS mice differ in NT-like immunoreactivity in specific brain regions, i.e., hypothalamus, and in NT receptor densities (Bmax) in frontal cortex and striatum. In other experiments LS and SS mice differed in behavioral responses to centrally administered NT. Intracerebroventricular (ICV) administration of NT produced dose-dependent changes in motor activity, hypothermia, and analgesia in both LS and SS mice. SS mice appeared to be more sensitive than LS to NT-induced analgesia but not hypothermia. Neurotensin increased or decreased locomotor activity in both SS and LS mice following intraventral tegmental area or ICV administration, respectively. The results indicate that LS and SS mice, which were selectively bred for differences in ethanol sensitivity, differ genetically in NT concentrations, receptor densities in specific brain regions, and in some receptor-mediated behavioral responses to NT. PMID- 2547209 TI - Pathogenicity and immunogenicity of mynah pox virus in chickens and bobwhite quail. AB - An avian pox virus was isolated from cutaneous proliferative lesions removed from greater hill mynahs (Gracula religiosa) imported from Malaysia. Cutaneous inoculation of specific pathogen-free chickens and bobwhite quail with the mynah pox virus resulted in severe proliferative cutaneous lesions similar to those seen in the naturally infected mynah birds. Microscopically, the reaction in the chickens and quail at sites of virus inoculation was characterized by marked epithelial hyperplasia with ballooning degeneration and formation of cytoplasmic inclusion bodies. Inoculation of conjunctival and oral mucosae of chickens by applying pox virus with a cotton swab did not result in gross or microscopic lesions. In cross-protection studies, chickens and bobwhite quail immunized with either quail, fowl, pigeon, turkey, or psittacine pox vaccines were not protected from challenge with mynah pox virus. Following vaccination of quail and chickens with mynah pox virus vaccine, there was no resistance to challenge by quail, fowl, pigeon, turkey, or psittacine pox viruses. Significant protection against development of lesions following inoculation with mynah pox virus was attained only when the homologous virus was used as a vaccine. PMID- 2547210 TI - Levels of dihomo-gamma-linolenate are depressed in heart phosphatidylcholine and phosphatidylethanolamine in the biotin-deficient chick. AB - Male broiler chicks were fed a purified diet with 0 or 500 micrograms/kg of added biotin for 21 days posthatch. Fatty acid composition of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were determined in erythrocytes and in heart mitochondria and microsomes of chicks at 12 and 21 days of age. The biotin deficient (BD, 0 microgram biotin) chicks had lower body weights, lower feed efficiencies, and higher scores for dermatitis and twisted leg than the biotin adequate (BA, 500 micrograms biotin) chicks. The erythrocyte arachidonate level in PC was higher in BD chicks compared to the BA controls, and the level increased with age in PC and PE in the erythrocytes. The level of dihomo-gamma linolenate (20:3 omega 6) in PC and PE in heart mitochondria and microsomes was consistently decreased in BD chicks compared with that in BA chicks. In heart subcellular organelles, the level of arachidonate in PC increased, but the level of dihomo-gamma-linolenate in PE decreased with age. These data demonstrate that biotin deficiency results in dramatic changes in the fatty acid composition of specific phospholipids in chick erythrocytes and heart subcellular organelles. PMID- 2547211 TI - The influence of soman on the membrane fluidity of rat superior cervical ganglion. PMID- 2547213 TI - Influence of long-term prenalterol treatment on the heart rate and the beta adrenoceptor binding-sites in rat myocardium. AB - The density of beta- adrenoceptors has been determined on membranes prepared from rat myocardium. Long-term administration of prenalterol resulted in an increase in the density of receptors, without any change in affinity. The average heart rate increase induced by intravenous prenalterol administration to denervated rats was, however, similar in the prenalterol pretreated and the control groups. We conclude that long-term treatment with a partial beta1-adrenoceptor agonist lead to an increased beta-adrenoceptor density in the myocardium. PMID- 2547212 TI - Effect of peripheral benzodiazepine receptor ligands on the contraction of isolated heart atrium and papillary muscle of rats. PMID- 2547214 TI - A combined in ovo-in vitro system for studies of volatile compounds on brain development: differential effects of carbon tetrachloride on neurones and astrocytes. AB - A method for studies of organic solvent effects on the development of neurones and astrocytes is described. Chick embryos in ovo were exposed to a low concentration of carbon tetrachloride vapours. After the exposure, primary cultures of neurones and astrocytes were prepared from the brain hemispheres of the embryos. Toxic effects of the solvent were analyzed in the cultures during the course of cellular differentiation as changes in cell growth and morphology, respiratory activity, and biochemical properties characteristic of each cell type. The development of neurones from embryos exposed from day 5 to day 8 to 25 p.p.m. of carbon tetrachloride was not affected by the exposure. On the other hand, growth rate and respiratory activity of astrocytes from embryos exposed from day 11 to day 14 to 25 p.p.m. of CCl4 were affected by the exposure, whereas no morphological changes or changes in the cyclic AMP levels could be observed. Consequently, low concentrations of carbon tetrachloride seem to affect the development of astrocytes rather than that of neurones in our experimental system. PMID- 2547215 TI - The role of brain serotonin in the electroconvulsive shock-induced changes in behavioural effects of intra-hippocampally injected clonidine. AB - The influence of central serotonin depletion upon behavioural effects of intra hippocampally injected clonidine in the electroconvulsive shock-treated rats (ECS), was studied. Repeated ECS significantly attenuated the depressive influence of clonidine upon the locomotor activity of the rats in the open field test. Chemical lesions to the median raphe nucleus (MR) did not significantly affect ECS-induced changes in clonidine activity in this test. In the forced swimming the MR lesions revealed the stimulatory potency of clonidine microinjections upon rat active behaviour. In animals pretreated with repeated ECS, clonidine also significantly potentiated swimming activity, but no evident synergism of ECS and MR lesion could be observed. Taking into account these and other data it is concluded that central serotonin depletion might differentially affect the adaptive processes occurring in the alpha 2-adrenoceptors in the course of treatment with tricyclic antidepressants and ECS, but it does not seem to be a strong phenomenon. Moreover, it is suggested that clonidine effects in the open field and forced swimming tests may be mediated by different neuronal substrates within the rat hippocampus. PMID- 2547216 TI - [Adjuvant therapy of non-small cell bronchial cancer with mopidamol]. AB - Between January 1982 and April 1986 a double-blind randomized placebo controlled study of mopidamol, employed as adjunct therapy to surgery in patients with non small cell bronchial carcinoma, was performed at 7 hospitals. The main criteria were occurrence of metastases and survival. Mopidamol was given perioperatively at a dose of 2 x mg i.v. daily, and postoperatively orally at a dose of 3 x 500 mg daily. The treatment period was scheduled to at least 2 years and in some of the patients was prolonged to 3 years. The standard therapy of each individual center was given as basic therapy. A total of 270 patients were included in the study, 147 in the placebo and 123 in the mopidamol group. By the end of the study 52 deaths from metastases had occurred in the placebo group (35%) compared with only 32 (26%) in the mopidamol group. This difference is statistically significant at p less than 0.05 with the one-sided test. A comparison of life tables according to Kaplan-Meier shows a statistically significant difference in favour of the group treated with mopidamol (savage p less than 0.05). Cox's multivariate analysis confirmed the statistically significant difference in favour of the group treated with mopidamol, the inclusion of the risk factors tumour stage and histology in the evaluation results in a p-value of 0.02. With respect to the incidence of metastases there were no appreciable differences between the treatment groups. The incidence of side effects or undesired events was equal in both groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547217 TI - Phospholipases C and A2 in malignant cell proliferation. PMID- 2547218 TI - Urocanic acid analogues and the suppression of the delayed type hypersensitivity response to Herpes simplex virus. AB - Ultraviolet irradiated urocanic acid (4-imidazoleacrylic acid) containing a mixture of cis- and trans-isomers has been shown previously to induce suppression of the delayed type hypersensitivity (DTH) response to Herpes simplex virus type 1 (HSV-1) in a murine model of infection. The cis-isomer of urocanic acid was prepared and the cis- and trans-isomers of 2-methylurocanic acid. 2 pyrroleacrylic acid, 2-furanacrylic acid, 2-thiopheneacrylic acid, 3 thiopheneacrylic acid as well as dihydrourocanic acid and histamine. Each was applied at concentrations of 1 and 50 micrograms per mouse to the shaved dorsal skin and the mice were infected subcutaneously with HSV 5 h later. After 8-10 days the DTH response to the virus was measured by an ear swelling test. It was found that cis-urocanic acid was effective in suppressing the DTH response at levels of 1 microgram per mouse or less. The cis- and trans-isomers of 2 furanacrylic acid, 2-pyrroleacrylic acid and 2-thiopheneacrylic acid were also effective, with the cis- form generally being more active than trans, and 2 pyrroleacrylic acid being particularly potent. Cis- and trans-3-thiopheneacrylic acid, on the other hand, were only marginally immunosuppressive while neither isomer of 2-methylurocanic acid had any suppressive ability. Dihydrourocanic acid and histamine were also shown to suppress the DTH response. Thus the structural features necessary for urocanic acid and its analogues to act as mediators of UV induced immunosuppression could be deduced and implications for their mechanism of action discussed. PMID- 2547219 TI - Beta adrenergic regulation of the hydrochloric acid secretion. AB - Propranolol induces basal achlorhydria in 30.7% of the patients, decreases significantly the HCl secretion in 42.3%, does not change it in 25% and increases it in 1.9%. After histamine stimulation (Kay test) propranolol decreases HCl in 57% and increases it in 43% of the cases. Propranolol induces a significant decrease of carbon anhydrase in the gastric mucosa. Serum gastrin is not significantly decreased after propranolol treatment. Isoprenaline increases the HCl secretion in all the patients investigated. Propranolol suppresses the effect of isoprenaline in 28.6% of the cases, decreases it in 33%, does not change it in 19% and increases it in 19%. The action of isoprenaline is not influenced by atropine, so the mechanism of action of the beta agonist is non-cholinergic. The gastric juice volume is not changed significantly by the blockage or stimulation of beta receptors. The beta adrenergic nervous system stimulates HCl secretion. PMID- 2547220 TI - Effects of ethanol in an open field apparatus: modification by U50488H and WIN 44441-3. AB - The effects of U50488H, a kappa agonist, and WIN 44441-3, a kappa antagonist, and their modification of the effects of ethanol, on the behavior of rats in a modified open field apparatus, was examined. Crossover activity was increased by U50488H. Headpoke activity was decreased by WIN 44441-3 and increased by U50488H. Rearing activity was increased by WIN 44441-3 but was not affected by U50488H. The effect of both drugs was dose related, with the largest doses having no effect. Ethanol (0.5 g/kg) stimulated crossover activity while it depressed rearing, headpoke and corner activities; except for crossover activity the 2.0 g/kg dose of ethanol depressed these activities. Pretreatment with WIN 44441-3 (0.5 mg/kg) potentiated the stimulant effect of ethanol on crossover activity and partially reversed the depressant effect of ethanol on rearing and headpoke activities. U50488H potentiated the ethanol-induced depression of headpoke and reversed the depression of corner activity. Pretreatment with U50488H had no effect on ethanol's action on crossover and rearing behaviors. Our results indicate that kappa opiate receptors may mediate some behaviors exhibited by rats in a modified open field apparatus. Activation of these receptors increases locomotor and headpoke activity but had no effect on rearing activity. Furthermore, the 0.5 g/kg dose of ethanol has differential effects on different measures of open field behavior, while the 2.0 g/kg dose was largely depressant. Our data suggest that some of these effects of ethanol may be mediated via kappa opioid receptors. PMID- 2547221 TI - Changes in forebrain Na,K-ATPase activity and serum hormone levels during sexual behavior in male rats. AB - We have previously shown that Na,K-ATPase activity (the enzymatic machinery for the sodium pump) in brain areas such as the medial basal hypothalamus (MBH) and the preoptic-suprachiasmatic region (POSC) can be changed by experimental manipulations of gonadal function. We now report enzyme levels in brain regions as related to hormonal changes occurring during sexual behavior. Male rats were exposed to receptive females and decapitated immediately after displaying one of the following behavioral events: the start of copulatory activity, first ejaculation, and the beginning of a second copulatory series. A group of noncopulating animals were used as control. The variables measured included serum levels of LH, PRL and testosterone and Na,K-ATPase activity in MBH, POSC and parietal cerebral cortex (CC). A steady increase in enzyme activity in the POSC, but not the MBH or CC, was found in copulating animals. Serum LH levels changed in a similar fashion. A sharp increase in serum PRL levels, seemingly related to ejaculation, was also observed. These data are consistent with our previous findings on monoaminergic neurotransmission in brain regions related to male sexual behavior. PMID- 2547222 TI - Effects of T-2 toxin on saccharin aversion and food consumption in adult rats. AB - The present experiment used a saccharin aversion paradigm to evaluate the potential aversive action of T-2 toxin, a trichothecene mycotoxin that induces emesis and weight loss. Adult male rats were fed either a control diet or a diet adulterated with 640 ppm lithium chloride (positive control) or with 2.5, 5.0 or 10.0 ppm T-2 toxin and given access to a 0.1% saccharin solution and to tap water during four training days. The rats were then shifted to the control diet during three extinction days. Moderate saccharin aversion induced by the positive control diet and the 5.0 and 10.0 ppm T-2 diets was apparent on the third day of exposure and the aversion to the saccharin solution abated during the extinction trials. Saccharin aversion was evident at levels of T-2 toxin that did not induce obvious tissue pathology. PMID- 2547223 TI - Pituitary-adrenal and thyroid effects on melatonin content of the rat pineal gland. AB - Based on clinical findings of diminished nocturnal serum melatonin levels in affective illness, we hypothesized that alterations in the pituitary-adrenal or thyroid axes of the rat might alter the nocturnal rise of melatonin content of the pineal gland in that species. Two experiments were conducted to investigate these issues. In the first, rats were injected for nine days with adrenocorticotropic hormone (ACTH) or corticosterone, timed to accentuate and prolong the normal circadian corticosterone rise. Although both these treatments produced significant elevations of serum corticosterone, there was no difference in pineal melatonin content during the day or night from that measured in control rats. In the second experiment, hypothyroidism was induced in rats by thyroid parathyroidectomy, and hyperthyroidism was produced by injection of triiodothyronine (T3) for nine days. Despite clear evidence of metabolic and endocrine effects of these thyroid manipulations, pineal melatonin content was not altered during the day or night. The nocturnal increase of melatonin may have been phase-advanced in the hypothyroid group, although the experiment was not designed to detect such a shift. There thus is no evidence from this study in the rat to suggest that diminished nocturnal melatonin production in affective illness might be due to associated alterations in the pituitary-adrenal or thyroid systems. PMID- 2547224 TI - Neuroendocrine responses to exogenous estrogen: no differences between heterosexual and homosexual men. AB - Plasma LH concentrations were determined in 55 men before and for four days following injection of 10 mg, 20 mg, or 30 mg Premarin or a placebo injection of vehicle. Testosterone (T) and dihydrotestosterone (DHT) concentrations were also determined in plasma samples taken just prior to Premarin or placebo injection and in samples taken three days later. All subjects provided self-descriptions of their sexual orientation and, based upon these descriptions, were classified as heterosexual (N = 39) or homosexual (N = 16). Premarin injection resulted in a reliable reduction in plasma LH 24 hr later. In subsequent samples, LH values rose and in many cases exceeded baseline levels, most reliably in those subjects receiving the 10 mg dose. Contrary to some previous reports, we observed no significant differences between heterosexual and homosexual subjects in the likelihood of their exhibiting elevated LH concentrations following exogenous estrogens. T, but not DHT, concentrations were suppressed after Premarin injection in both groups of subjects. Other than Premarin dosage, we could not identify any variable which predicted the likelihood of elevated LH values. PMID- 2547225 TI - Spontaneous exploration of a 6-arm radial tunnel maze by basal forebrain lesioned rats: effects of the benzodiazepine receptor antagonist beta-carboline ZK 93 426. AB - Nine days following ibotenic acid induced basal forebrain lesions or a sham operation, rats were allowed to explore an automated six-arm radial tunnel maze. From each session, several measures of locomotor and exploratory activity were registered. Lesioned and sham-operated animals were treated with either the benzodiazepine receptor antagonist beta-carboline ZK 93 426 (5 mg/kg; IP) or vehicle (Cremofor EL 10% in saline; IP; n = 10 for each group). Treatment was carried out 30 min before each session during acquisition (seven sessions) and reversals of the maze configuration (seven sessions). Eight days following the 14th session, the animals were retested without any further drug treatment. The main results suggest that the lesion resulted in locomotor hyperactivity, an increase in the number of blind arm entries, and of choice stereotypy. Treatment with ZK 93 426 attenuated the lesion-induced alterations of locomotor and exploratory activities. During the retest, the lesioned, previously vehicle treated rats revisited arms which they had already explored during this session more frequently than the lesioned, previously ZK-treated rats; the latter group did not differ from the sham-lesioned controls. It is concluded that basal forebrain lesioned animals explored the tunnel maze less efficiently than sham lesioned controls and that the lesioned animals benefited from the treatment with ZK 93 426. Although the specificity of the lesion in terms of destruction of cholinergic neurons remains unsettled, and although the psychological significances of the behavioral measures obtained from the tunnel maze are not yet fully understood, these results suggest that antagonists or partial inverse agonists at the benzodiazepine receptor may be able to normalize basal forebrain lesion-induced behavioral alterations. PMID- 2547226 TI - Neurokinin A, neurokinin B and neuropeptide K in the rabbit iris: a study comparing different extraction methods. AB - Neurogenic inflammation in the rabbit eye is thought to be partly mediated by tachykinins released from trigeminal sensory nerve fibres. In the present study we have investigated the occurrence of neurokinin A (NKA), neurokinin B (NKB), neuropeptide K (NPK) and related immunoreactive components in the rabbit iris ciliary complex using neutral and different types of acidic media for extraction, reverse phase high performance liquid chromatography (RP-HPLC) and radioimmunoassay (RIA). The immunoreactive material detected with an antiserum reacting almost equally well with NKA, NKB and NPK consisted mainly of NKA, and small amounts of NPK but almost no NKB. Acidic media seemed to be more effective than neutral media for extraction of NKA and NPK. Acid extraction yielded also an NKA-immunoreactive component which eluted immediately before NKA while neutral extracts, on the other hand, contained a component which appeared behind NKA, i.e. in the position of NKA-(3-10) and NKA-(4-10). The present results indicate that NKA but not NKB may play a role in neurogenic inflammation in the rabbit eye. PMID- 2547227 TI - The effects of hemorrhagic shock on thyrotropin-releasing hormone and its receptors in discrete regions of rat brain. AB - The effects of hemorrhagic shock on thyrotropin-releasing hormone (TRH) levels and its receptors were studied in different regions of the rat brain. Rats were bled for 30 min from the left femoral artery, and their mean arterial pressure was kept at 40 mmHg for the following hour. The rats were killed by decapitation. Rat brains were immediately removed and dissected into 7 regions. Hemorrhagic shock decreased TRH significantly in the frontal cortex, septum, hippocampus, and hindbrain but TRH was not changed in the striatum, hypothalamus, and midbrain. Hemorrhagic shock significantly decreased TRH receptor binding in the septum and hindbrain. Scatchard analysis of saturation isotherms of specific TRH binding showed that the decreased specific TRH binding in the hindbrain resulted not from an increase of the dissociation constant (Kd), but from a decrease in the maximum number of binding sites (Bmax). In the septum, the decrease in specific binding was due both to a decrease in Bmax and an increase in Kd. The findings indicate that TRH plays a role in the physiological response to hemorrhagic shock. PMID- 2547228 TI - [Renal scintigraphy in the making of customized shields in radiotherapy of the upper abdomen]. PMID- 2547229 TI - [A case of chondrosarcoma developing within the metastasis of germ cell tumor of the testis]. PMID- 2547230 TI - [HLA and cytomegalovirus infection in pregnant women]. PMID- 2547231 TI - [Chronic adrenal failure secondary to an isolated deficiency of ACTH. Presentation of 2 new cases]. AB - Chronic adrenal failure due to hypothalamic-hypophyseal disorders is rarely encountered. This can be due to hormonal deficiencies generally related to the presence of a brain tumor or an infiltrating process and sometimes to an isolated deficiency of ACTH. We report on two patients with adrenal failure with low basal cortisol levels and a poor response to short cortisol stimulation test with ACTH. Long cortisol stimulation test with ACTH was normal and ACTH was not stimulated with corticotropin releasing factor. However, other specific dynamic pituitary hormonal tests were normal. The uncommon clinical presentation of the disease, such as severe hypoglycemic crisis and fever of unknown origin (FUO), is underscored. PMID- 2547233 TI - [Cutaneous metastasis as the first manifestation of hepatocarcinoma]. PMID- 2547232 TI - [Severe polyneuropathy as the presenting form of idiopathic hypereosinophilic syndrome]. PMID- 2547234 TI - Expression of herpes simplex virus thymidine kinase, mediated by vaccinia virus early promoters. AB - We measured herpes virus thymidine kinase (HSV-TK) activity in extracts from cells infected with eight vaccinia virus recombinants (VpT), each expressing HSV TK under the control of an early promoter previously isolated by a shotgun procedure. The HSV-TK activities induced by the VpT recombinants were compared to that produced under the control of the vaccinia virus thymidine kinase (VV-TK) promoter in VMM5-TK recombinant virus. The insert from VpT38 was approximately 10 times more efficient than the VV-TK promoter for HSV-TK expression, reflecting a similar relative strength of the promoters. HSV-TK activities induced by the other VpT recombinants varied between one and ten times that expressed by the VV TK promoter, but the nucleotide sequence of the 5'-end region of their mRNA suggested that these values did not necessarily reflect the strength of corresponding promoters. No significant reduction in HSV-TK activity was noted for two VpT and the VMM5-TK recombinants when viral DNA replication was prevented, but a significant reduction (30 to 75%) was observed for the other six recombinants studied. These results suggested that some early genes of vaccinia virus are expressed only during the early stage of infection, whereas others continue to be expressed at the late stage. The strength of two vaccinia early promoters (VpT38, PF) relative to that of the VV-TK promoter was deduced from HSV TK activities induced by comparable vaccinia virus recombinants. PMID- 2547235 TI - Effect of alkaloids isolated from Amaryllidaceae on herpes simplex virus. AB - Studies were carried out on the effects of Amaryllidaceae alkaloids and their derivatives upon herpes simplex virus (type 1), the relationship between their structure and antiviral activity and the mechanism of this activity. All alkaloids used in these experiments were biosynthesized from N benzylphenethylamine; the apogalanthamine group was synthesized in our laboratory; those which may eventually prove to be antiviral agents had a hexahydroindole ring with two functional hydroxyl groups. Benzazepine compounds were neither cytotoxic nor antiviral, but many structures containing dibenzazocine were toxic at low concentrations. It was established that the antiviral activity of alkaloids is due to the inhibition of multiplication and not to the direct inactivation of extracellular viruses. The mechanism of the antiviral effect could be partly explained as a blocking of viral DNA polymerase activity. PMID- 2547236 TI - Evaluation of the induction of specific cytotoxic T lymphocytes following immunization of F1 hybrid mice with rabies antigens. AB - The measurement of the production of cytotoxic T lymphocytes (CTL) induced in mice by rabies antigens currently uses spleen cells from immunized A/J mice as effector cells and infected neuroblastoma syngeneic cells as target cells. For several reasons, including difficulties in obtaining A/J mice, as well as genetic analysis of immune responses, it would be advantageous to use strains of mice other than the A/J mice. However, cell lines other than the neuroblastoma Neuro 2a line are difficult to infect by the rabies virus. Therefore, using the same target cells expressing the major histocompatibility complex H-2kd, we have developed an experimental system based on the induction of CTL in F1 BALB/c X C3H/HeJ hybrid H-2kd mice. Splenocytes from F1 hybrid mice primed with inactivated purified rabies virus (IPRV) exhibited cytotoxic activity specific for syngeneic infected target cells (Neuro-2a). High amounts of IPRV were required for the induction of CTL following in vivo priming. The antigen dose required for CTL induction was reduced by in vitro restimulation. In addition to specific CTL, a high level of natural killer cell activity was induced in F1 hybrid mice by priming with IPRV. Among rabies antigen preparations tested (IPRV, purified glycoprotein and ribonucleoprotein), only IPRV induced strong CTL stimulation. PMID- 2547237 TI - Reactogenicity and immunogenicity of rotavirus WC3 vaccine in 5-12 month old infants. AB - Rotavirus is a major cause of acute gastroenteritis in infants worldwide and there is need for an effective vaccine. Rotavirus Wistar calf 3 (WC3) is a strain of bovine origin attenuated by 12 passages in cell culture. A lyophilized candidate vaccine containing 1 x 10(7) PFU of WC3 has been developed. An oral dose was given to 25 French infants 5-12-months old (mean age 8.6 months). No diarrhoea was observed within 2 weeks after vaccination. Unexplained vomiting was reported once and isolated fever greater than 37.8 degrees C was reported 3 times during the first week. One month later, a neutralizing antibody response to serotypes tested was shown in 88% of cases, with heterotypic responses to human serotype 3 (SA11 strain) in 72% and to type 1 (WA strain) in 48%. The percentage of immune response was similar whether the infant had antibody prior to immunization or not, but a booster effect was observed in children who had pre immunization rotavirus antibodies. Considering these promising results, efficacy trials are in the planning in different parts of the world. PMID- 2547239 TI - The aging neuromuscular system. PMID- 2547238 TI - In vitro sensitivity of human herpesvirus-6 to antiviral drugs. AB - We studied the sensitivity of human herpesvirus-6 (HHV-6) to 4 antiviral drugs known to be effective in the treatment of infections with other human herpesviruses and human immunodeficiency virus. HHV-6 was grown in peripheral blood lymphocytes, and virus multiplication was quantified by evaluation of the cytopathic effect by molecular hybridization and indirect immunofluorescence assay. The 50% and 90% inhibitory concentrations (IC50 and IC90) were determined for each drug. The results obtained by the 3 different quantification techniques were found to correlate, and enabled us to conclude that HHV-6 replication was readily inhibited by foscarnet or ganciclovir. In contrast, inhibition of HHV-6 replication was observed only at high concentrations of acyclovir, and was not detected at the tested concentrations of zidovudine. PMID- 2547240 TI - [Instinct therapy--raw food with meat with exclusion of milk products. Report No. 16]. AB - Instinct therapy is a diet consisting exclusively of food in its natural form, which has not been subjected to cooking, preserving or seasoning. Dairy products are forbidden. Indications are numerous and rather imprecise including especially cancer. Burger developed Instinct therapy in 1964 as a diet according to his own philosophy. 1980 he left Switzerland because the practice of "metasexuality" had caused him problems with the law. He founded a school of Instinct therapy in 1982 in France and published his book "La guerre du cru" (The Raw Food Fight) in 1985. In Switzerland, Besuchet took on the teaching of Burger in 1978 and was very active in giving lectures. He died in June 1985 of mediastinal cancer. According to Burger, man originally ate raw food and his instinct has not evolved since those pre-historic times, but has been "artificially modified" by cooked food and the consumption of non-human milk. If man could re-develop his natural instinct and eat only fresh food, which is not prepared in any way, he could solve his health problems and restore defenses against diseases such that these could become beneficial. Cooked food, milk and cereals are thought to be the cause of cancer but instinctive eating would help to avoid or to cure cancer. No preclinical or clinical trials have been carried out. The theories of Burger and Besuchet are based on unverifiable personal experiences. PMID- 2547241 TI - [Cutaneous leishmaniasis]. PMID- 2547242 TI - Cytomegalovirus the predominant cause of pneumonia in renal transplant patients. A two-year study of pneumonia in renal transplant recipients with evaluation of fiberoptic bronchoscopy. AB - The microbiological etiology of pneumonia in 34 renal transplant patients with clinical and X-ray evidence of pulmonary parenchymal disease was studied. Fiberoptic bronchoscopy with bronchoalveolar lavage (BAL), transbronchial lung biopsy (TBB) and brushing was performed on 18 patients. Laboratory evaluation included histological and cytological methods, cultures for bacteria, fungus and virus and immunofluorescence techniques for the detection of Pneumocystis carinii, cytomegalovirus (CMV) and legionella. Serum samples were obtained concomitantly for antibody studies. CMV, the most common etiology, was considered to be the cause of disease in 18/34 patients. All but one of these patients had positive CMV isolates in culture on leucocytes. Pulmonary edema was found in 7 patients, bacterial pneumonia in 11 patients, P. carinii in 4 patients and Candida albicans in 1 patient. Multifactorial etiology was found in 12/34 cases. The overall mortality was 32%. Bronchoscopy gave correct diagnosis in 13/14 patients with infectious pulmonary diseases (93%). Bronchoscopy procedures were well tolerated and should be considered in transplant patients with evidence of pulmonary parenchymal disease. PMID- 2547243 TI - Varicella zoster virus deoxythymidine kinase is present in serum before the onset of varicella. AB - A sensitive enzyme assay with 125I-iododeoxyuridine as substrate and cytidine triphosphate as phosphate donor was used for the direct detection of varicella zoster virus (VZV) deoxythymidine kinase (TK) in human serum. Sera sampled during the incubation period of varicella from 2 patients, a 42-year-old man and his 11 year-old son, have been analysed for TK activity. A simultaneous increase in cellular and VZV TK activity, starting 5 to 3 days before the onset of clinical varicella, was observed. PMID- 2547244 TI - New triazole antifungal agents (fluconazole and itraconazole) in the treatment of HIV-related gastrointestinal candidiasis. PMID- 2547245 TI - A comparative study of ciprofloxacin and conventional therapy in the treatment of patients with chronic lower leg ulcers infected with Pseudomonas aeruginosa or other gram-negative rods. AB - Twenty-six elderly patients with chronic leg ulcers infected by Pseudomonas aeruginosa or other aerobic Gram-negative rods were randomised to two treatment groups. The control group (eight patients) received conventional local therapy and the other group (18 patients) was treated with oral ciprofloxacin for three months in addition to conventional local therapy. In the beginning of the study both groups were comparable with the age of the patients and the associated diseases including impairment of arterial and venous circulation in the lower legs. Also the size, duration and the severity of the inflammation reaction in the leg ulcers were comparable before the start of the therapy. Ciprofloxacin was clinically more effective than the standard therapy in reducing the size of the ulcer (p less than 0.05). Also the need of extra systemic antibiotics decreased significantly in the ciprofloxacin group compared with the controls. In three out of eighteen ciprofloxacin treated patients the leg ulcers disappeared completely during the three months' study period compared with none in the control group. However, ciprofloxacin resistant strains, mainly staphylococci, appeared in the leg ulcers in 67% of the ciprofloxacin treated patients compared with 0% in the control group (p less than 0.01). No significant side-effects due to ciprofloxacin except the resistant strains were noticed. We conclude that oral long-term ciprofloxacin therapy is effective in the treatment of chronic leg ulcer infections due to Gram-negative rods but selection of ciprofloxacin resistant strains is a problem in this patient group. PMID- 2547246 TI - Renal parenchymal damage on DMSA-scintigraphy in pelviureteric obstruction. AB - During a 1.5 year period 21 children were investigated with 99-m-technetium dimercaptosuccinic acid (DMSA) before operation for hydronephrosis due to pelviureteric obstruction. The age at investigation was 0.2-11.5 years. Fourty two kidneys were examined. Hydronephrosis existed on the right side in 8 cases, left side in 9 and bilateral in 4 cases. Seventeen kidneys had no obstruction. The scintigraphy was interpreted as normal in 19 kidneys. Decreased isotope uptake was found in 23 kidneys and localized to the upper pole area in 19 kidneys, middle-lateral part in 7, lower pole area in 15 and the middle-medial part in 12 kidneys. There were no predominance for any part of the kidney to be affected by parenchymal damage. In 8 children investigated before the age of 1 year, 4 of 10 hydronephrotic kidneys revealed normal DMSA scintigram. DMSA scintigraphy delineates functioning renal parenchyma. DMSA scintigraphy delineates functioning renal parenchyma. It can be recommended as a routine method for evaluation of the renal parenchyma before surgery and for follow up studies in all ages of childhood. PMID- 2547247 TI - Recombinant 47-kilodalton cytosol factor restores NADPH oxidase in chronic granulomatous disease. AB - A 47-kilodalton neutrophil cytosol factor (NCF-47k), required for activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase superoxide (O2-.) production, is absent in most patients with autosomal recessive chronic granulomatous disease (AR-CGD). NCF-47k cDNAs were cloned from an expression library. The largest clone predicted a 41.9-kD protein that contained an arginine and serine-rich COOH-terminal domain with potential protein kinase C phosphorylation sites. A 33-amino acid segment of NCF-47k shared 49% identity with ras p21 guanosine triphosphatase activating protein. Recombinant NCF-47k restored O2-. -producing activity to AR-CGD neutrophil cytosol in a cell-free assay. Production of active recombinant NCF-47k will enable functional regions of this molecule to be mapped. PMID- 2547248 TI - Beta-adrenergic inhibition of cardiac sodium channels by dual G-protein pathways. AB - The signaling pathways by which beta-adrenergic agonists modulate voltage dependent cardiac sodium currents are unknown, although it is likely that adenosine 3'5'-monophosphate (cAMP) is involved. Single-channel and whole-cell sodium currents were measured in cardiac myocytes and the signal transducing G protein Gs was found to couple beta-adrenergic receptors to sodium channels by both cytoplasmic (indirect) and membrane-delimited (direct) pathways. Hence, Gs can act on at least three effectors in the heart: sodium channels, calcium channels, and adenylyl cyclase. The effect on sodium currents was inhibitory and was enhanced by membrane depolarization. During myocardial ischemia the sodium currents of depolarized cells may be further inhibited by the accompanying increase in catecholamine levels. PMID- 2547250 TI - Response of guinea pigs to intravaginal inoculation with guinea pig cytomegalovirus. AB - Female guinea pigs were inoculated intravaginally with guinea pig cytomegalovirus (GPCMV) propagated either in guinea pig embryo fibroblast cultures (GPEF) or salivary glands. The incidence of infection was higher with GPEF virus. Rare instances of isolation of GPCMV from cervical swabs 9-48 hr after inoculation was attributed to survival of inoculum in the genital tract. Neither immunofluorescence microscopy nor histopathologic examination showed evidence for active infection of genital tract tissue examined up to day 5 after inoculation. At necropsy on days 30-49, GPCMV was isolated from salivary glands and occasionally from pancreas and lymph nodes. Seroconversion following intravaginal inoculation was demonstrated by an enzyme-linked immunosorbent assay (ELISA) test, and titers were comparable to those after intraperitoneal or subcutaneous inoculation. However, titers of neutralizing antibody, determined by plaque reduction assay, were significantly lower in the group inoculated intravaginally. These findings are relevant to consideration of cytomegalovirus as a sexually transmitted agent in humans. PMID- 2547249 TI - 5-bromo-2'-deoxyuridine blocks myogenesis by extinguishing expression of MyoD1. AB - The pyrimidine analog 5-bromodeoxyuridine (BUdR) competes with thymidine for incorporation into DNA. Substitution of BUdR for thymidine does not significantly affect cell viability but does block cell differentiation in many different lineages. BUdR substitution in a mouse myoblast line blocked myogenic differentiation and extinguished the expression of the myogenic determination gene MyoD1. Forced expression of MyoD1 from a transfected expression vector in a BUdR-substituted myoblast overcame the block to differentiation imposed by BUdR. Activation of BUdR-substituted muscle structural genes and apparently normal differentiation were observed in transfected myoblasts. This shows that BUdR blocks myogenesis at the level of a myogenic regulatory gene, possibly MyoD1, not by directly inhibiting the activation of muscle structural genes. It is consistent with the idea that BUdR selectively blocks a class of regulatory genes, each member of which is important for the development of a different cell lineage. PMID- 2547251 TI - [Omega-exonuclease: an enzymatic marker for basophilic leukocytes and mast cells]. AB - The activity of the enzymatic marker for basophilic leucocytes and mast cells, omega-exonuclease, was assayed in 20 peripheral blood and 10 bone marrow samples from haematologically normal subjects. The absolute specificity and technical simplicity, of this cytochemical method were fully confirmed. PMID- 2547252 TI - Repeat hepatectomy for recurrent malignant tumors of the liver. AB - The results of 34 repeat hepatectomies for recurrent malignant tumors of the liver in 28 patients are reported herein. There was no operative mortality, and the morbidity rate was 15 per cent. No instances of postoperative hepatic insufficiency were observed. Only five of the repeat hepatectomies were extensive (three or more hepatic segments). Thirteen resections were performed upon 11 patients for recurrent hepatocellular carcinoma. Four of these patients are still alive, one patient with a known recurrence (a mean survival time of 33 months with a range of four to 54 months). Ten resections were performed upon nine patients for recurrent metastases from the colon and rectum. Five of these patients are alive, four without apparent recurrence (a mean survival time of 13 months and a range of one to 35 months). Eleven resections were performed upon eight patients for recurrent miscellaneous malignant tumors, and four patients are alive, of whom three have a recurrence. The results of this study demonstrate that some patients benefit from repeat hepatectomy for recurrence of malignant hepatic tumors. Repeat hepatectomy is technically highly feasible. Economic hepatic resection with a sound oncologic basis (segmentectomy) is the operation of choice. PMID- 2547253 TI - Hippocampal unit activity after transient cerebral ischemia in rats. AB - Single unit activity of CA1 and CA3 neurons in the hippocampus was recorded in rats 1, 2, or 3 days after 10 minutes of transient cerebral ischemia induced by the clamping of both carotid arteries combined with hypotension. In addition, paired pulse inhibition/facilitation of the CA1 population spike was examined on Day 2 using two successive stimuli of the contralateral CA3 region delivered at various intervals. On Day 1, the mean +/- SEM firing rate in the CA1 region was 0.91 +/- 0.42/sec (n = 5), which was not significantly different from the control value of 0.98 +/- 0.26/sec (n = 5). Firing rate increased on Days 2 and 3 to 3.96 +/- 0.69/sec (n = 5), and 6.49 +/- 0.89/sec (n = 5), respectively. In the CA3 region, the mean +/- SEM firing rate of 1.18 +/- 0.27/sec in the five control rats sharply dropped to 0.14 +/- 0.11/sec in the five Day 1 rats and gradually increased to 0.45 +/- 0.11/sec in the five Day 3 rats. Histologic examination of these rats revealed ischemic changes restricted to CA1 neurons on Days 2 and 3. The paired-pulse experiment showed no significant difference between six control and six Day 2 rats in the inhibition of the second population spike with interstimulus intervals of less than 400 msec. At interstimulus intervals of greater than 500 msec there was facilitation of the second spike, which lasted 5 seconds in Day 2 rats. This facilitation was not observed in control rats. Because CA3 neurons constitute the main input to CA1 pyramidal cells, decreased activity of CA3 neurons indicates less excitatory input to CA1 neurons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547254 TI - Intravenously and iontophoretically administered naloxone reverses ischemic changes in rat hippocampus. AB - Forty rats under urethane anesthesia were subjected to cerebral ischemia by ligation of the right carotid, the right plus the left carotid, or the right carotid plus two vertebral arteries. Ischemia caused three types of changes in the field potential of the right hippocampal CA1 region evoked by fimbrial stimulation: 1) completely reversible deterioration (57% and 16% of the rats with unilateral and bilateral carotid artery ligation, respectively), 2) moderate deterioration (37% and 24% of the rats with unilateral and bilateral carotid artery ligation) and 3) irreversible loss of the evoked activity (6% and 60% of the rats with unilateral and bilateral carotid artery ligation and all the rats subjected to three-vessel occlusion). Naloxone improved the moderate deterioration in 10 of 11 rats (1-3 mg/kg i.v.) and in 15 of 16 (50-150 nA) iontophoretic applications, but naloxone did not restore the lost evoked activity. Intravenous morphine (10 mg/kg) aggravated the ischemic changes, and this effect was reversed by naloxone, while iontophoretic administration of morphine caused only excitation. These findings suggest that naloxone has a favorable effect on cerebral ischemia not severe enough to cause transmission failure. The reversal of ischemic changes by iontophoretic naloxone indicates that its site of action is at the neuronal or microcirculatory level. PMID- 2547255 TI - The mechanism of action of retrograde oxygen persufflation in renal preservation. AB - Vascular perfusion of gaseous oxygen has been used to prolong the in vitro survival of a number of isolated organs, and has been shown to improve the hypothermic preservation of ischemically injured kidneys that were subsequently transplanted. We have investigated the mechanism of this effect. Rabbit kidneys were subjected to 60 min of warm ischemia prior to preservation for 24 hr with Ross, Marshall, and Escott's hypertonic citrate solution, with or without retrograde oxygen persufflation (ROP) via the renal vein. It was found that adenine nucleotide levels were almost doubled in the ROP-preserved kidneys, principally due to higher adenosine triphosphate (ATP) concentrations. It was shown that cytochrome oxidase activity was unaffected by ischemia or preservation method, but studies with the metabolic inhibitors ouabain and a mixture of cyanide and iodoacetate suggested that ATP was being synthesized during the storage period but was also being utilized to power the active volume-regulating pump. Morphological examination revealed a much greater degree of cell swelling and cytological injury in the kidneys not subject to ROP, and the interstitial space appeared much reduced in the latter group. At the ultrastructural level, the ROP-treated kidneys showed generally well-preserved mitochondria, mostly in the energized "orthodox" configuration. In contrast, the mitochondria in the nonpersufflated kidneys were generally in the "condensed" deenergized state. We conclude that the provision of sufficient oxygen by ROP allows the continued production of ATP in sufficient quantities to permit improved maintenance of cellular volume and morphology under the conditions of low-temperature storage that we have studied. PMID- 2547256 TI - Prevention of cytomegalovirus-related death by passive immunization. A double blind placebo-controlled study in kidney transplant recipients treated for rejection. AB - In a double-blind placebo-controlled study, the value of prophylactic anti-CMV immunoglobulin administration was evaluated in 39 kidney transplant recipients treated for rejection with rabbit antithymocyte globulin. Passive immunization completely prevented CMV-related death, although it did not reduce the incidence of CMV isolation, viremia, or disease. The effect of passive immunization was exclusively observed in CMV-seronegative recipients of a CMV-seropositive kidney donor. It could be demonstrated even when instituted when antirejection therapy was started. Seropositive recipients did not benefit from immunoglobulin treatment. Moreover, CMV-seronegative recipients of a kidney from a seronegative donor were not at risk for CMV infection at all. Therefore passive immunization should be restricted to seronegative recipients of seropositive allograft donors treated for rejection. PMID- 2547257 TI - Cytomegalovirus infection in pediatric liver recipients. A virological survey and prophylaxis with CMV immune globulin and early DHPG treatment. AB - From November 1985 to August 1987, 22 children underwent orthotopic liver transplantation (OLT). Primary cytomegalovirus (CMV) infection was observed in 3 of 10 seronegative recipients (30%). It was severe in 2 cases, which were treated successfully by a combination of DHPG and CMV hyperimmune globulin. From September 1987 to August 1988, a virologic survey was undertaken. It enabled us to take prophylactic measures and to make an early diagnosis of CMV infection, permitting early antiviral treatment. During this period, 26 children received liver grafts. Nine of 13 seronegative recipients (69, 2%) developed primary CMV infection. Seven had received immunoprophylaxis and 8 were treated by DHPG. None had severe disease and all improved rapidly. PMID- 2547258 TI - Neoadjuvant chemotherapy and orthotopic liver transplantation for hepatocellular carcinoma. PMID- 2547259 TI - The molecular biology of hepatitis B virus. AB - Infection with the hepatitis B virus (HBV) or related hepadnaviruses is associated with a wide spectrum of liver diseases, including hepatocellular carcinoma (HCC). In this article we review the current state of knowledge about the structure, genetic organization and life cycle of HBV. The mechanisms of viral pathogenesis and HCC development remain poorly understood, but new approaches may soon begin to shed light on these areas. PMID- 2547260 TI - [Genital herpes]. PMID- 2547262 TI - [Ability of bentonite and natural zeolite to adsorb aflatoxin from liquid media]. AB - The adsorption of aflatoxin (AF) B1, contained in aqueous medium or in synthetic medium left after the cultivation of Aspergillus parasiticus NRRL 2999, was studied in two samples of bentonite, two samples of natural zeolite, and three kinds of adsorption coal added to water, to saline, to the blood serum of pigs, to the stomach fluid of pigs or rumen fluid of cows at concentrations of 5 to 50 mg per litre. Unlike retinol-propionate and beta-carotene, AF was readily adsorbable in vitro. The initial concentration of 3.6 mg or 18 mg AFB1 per litre was reduced to 0.3-27% after exposure to 50 g of adsorbent; this reduction was greater when the AFB1 was tested in the cultivation medium and the maximum reduction was recorded when adsorption coals was involved. The effectiveness of zeolite was lower in media containing nitrogen compounds and in those cases when its doses were low. The average AF retention in the adsorbents, calculated after the determination of desorption by chloroform extraction, was 72% of the original content in the medium in the case of adsorption coals, 66% in bentonite, and 60% in zeolite. PMID- 2547261 TI - [Regulation of the immune response using leukotrienes]. AB - Leukotrienes are derived from lipoxygenation in position 5 of arachidonic acid. They have numerous biological activities, mainly in inflammatory processes. During the last few years, however, it has become quite evident that they also possess a potential for immunoregulation. In particular, LTB4 can induce suppressor cells, activate the synthesis and production of lymphokines such as interleukin 2 or interferon-gamma, stimulate the production of interleukin 1 by monocytes or macrophages, and activate cytotoxic cells against virus-infected or tumor target cells. These findings suggest that LTB4 can play an important role in the interaction between inflammatory and immune phenomena. PMID- 2547263 TI - [Occurrence of viral particles of avian sarcoma viruses in sarcoma tissue]. AB - An electron-microscopic examination was performed of chicken fibrosarcoma caused by avian sarcoma virus (ASV), strain B 77, to investigate virus budding and release through the cytoplasmic membrane. The virus particles of type C- were 90 100 nm in size, the electron-optically denser nucleoids being clearly differentiated from the outer membrane. PMID- 2547264 TI - Haematological changes in two thoroughbred horses in training with confirmed equine herpesvirus 1 infections. AB - An outbreak of respiratory disease among thoroughbred horses in training in Hong Kong was caused by equine herpesvirus 1 (EHV-1) subtype 1 (abortion strain). Two of the horses affected by EHV-1 were serially blood sampled over a period of several weeks and their haematological values measured. There was an increase in monocyte count in the first few days which steadily decreased in one horse, but the other had a second monocyte peak after a period of exercise, thus demonstrating the importance of not working animals in the early stages of the disease. PMID- 2547265 TI - Occurrence of equine infectious anaemia in India. PMID- 2547266 TI - Equine herpesvirus. PMID- 2547267 TI - Diagnosis of FIV infection. PMID- 2547268 TI - Porcine Clostridium perfringens type A spores, enterotoxin and antibody to enterotoxin. AB - Forty-two Clostridium perfringens type A strains isolated from cases of diarrhoea in pigs were tested for their ability to sporulate and produce enterotoxin in three different sporulation media. Enterotoxin was produced by 11 of the 42 C perfringens type A isolates (26.2 per cent). Thirteen isolates (30.9 per cent) produced spores at a frequency of 10 per cent or more. Spore production was recorded in 24 (57.1 per cent) of the isolates. The titres of enterotoxin produced by the isolates ranged from 1:2 to 1:64. The enterotoxin produced was compared with that produced by a reference strain and found to be identical. Ninety-eight of 106 sow sera from four different farms were found to possess antibodies to C perfringens type A enterotoxin with titres ranging from 1:2 to 1:64. Spores of C perfringens type A were detected in pig faeces and intestinal contents in 20 of 23 cases of enteritis at levels of up to 5 x 10(6) cells/g of faeces. Smaller numbers of spores, up to 2 x 10(4)/g were present in five of 10 samples from non-diarrhoeic pigs. Enterotoxin was demonstrated by Vero cell assay in five of the 23 samples from diarrhoeic pigs but in none of the 10 samples from non-diarrhoeic animals. It was clear from these studies that C perfringens type A strains in pigs could sporulate and produce enterotoxin in vitro and in vivo and that enteritis might be associated with sporulating organisms in vivo. PMID- 2547270 TI - [Principles for prevention, early detection, treatment and after-care of malignant neoplasms of the trachea, bronchus and lung]. PMID- 2547269 TI - [Effect of corticotropin and glucocorticoids on lipid metabolism (a review of the literature)]. PMID- 2547271 TI - [Joint position paper of the directors of the Pneumonophthisiological Society of Czechoslovakia and the Society for Pulmonology and Tuberculosis of the German Democratic Republic on the problems of ambulatory pulmonology in the care of patients with bronchial carcinoma]. PMID- 2547272 TI - [The frequency and localization of hematogenic distant metastases of bronchial carcinoma]. AB - We examined on the basis of 289 autopsies the localization and frequency of metastases in 383 patients with bronchogenic carcinoma. In correspondence with the literature the main localisations of metastases were found in liver, suprarenal glands and skeleton. The small cell carcinoma showed the highest metastatic frequency, especially with 65.9% of liver metastases. The evidence of liver metastases in the phase before treatment is very important, because of the frequency of combination of liver metastases with metastases of suprarenal glands and skeleton. PMID- 2547273 TI - The interstitial system of the spinal trigeminal tract in the rat: anatomical evidence for morphological and functional heterogeneity. AB - Utilizing cyto-, myelo-, and chemoarchitecture as well as connectional criteria, the present study reveals the interstitial system of the spinal trigeminal tract (InSy-SVT) in the rat to be composed of five morphologically and functionally distinct components that are distributed within spatially restricted regions of the lateral medulla. The first component is represented by scattered interstitial cells and neuropil, which extend laterally into SVT from the superficial laminae of the medullary dorsal horn (MDH). The second component, the dorsal paramarginal nucleus (PaMd), consists of a small group of marginal (lamina I)-like neurons and neuropil situated within the dorsolateral part of SVT at the rostral pole of MDH. The third component represents a trigeminal extension of the parvocellular reticular formation (V-Rpc) into the ventromedial aspect of SVT at levels extending from rostral MDH to the caudal part of trigeminal nucleus interpolaris (Vi). The fourth component, the paratrigeminal nucleus (PaV), consists of a large accumulation of neurons and neuropil situated within the dorsal part of SVT throughout the caudal half of Vi. The fifth component is the insular trigeminal cuneatus lateralis nucleus (iV-Cul), which is a discontinuous collection of neurons and neuropil interspersed among fibers of SVT as well as wedged between it and the spinocerebellar tract. Thalamic projection neurons are located in PaMd and V-Rpc, whereas cerebellar projecting neurons are confined to iV-Cul. PMID- 2547274 TI - Putative nociceptive modulatory neurons in the rostral ventromedial medulla of the rat display highly correlated firing patterns. AB - Recent work in this laboratory has identified two classes of putative nociceptive modulating neurons in the rostral ventromedial medulla (RVM) of the rat: "off cells," which pause beginning just prior to the tail flick response (TF) evoked by noxious heat, and "on-cells," which accelerate shortly before the occurrence of the TF. In the unstimulated, lightly anesthetized rat, the spontaneous firing pattern of individual on- and off-cells consists of alternating periods of silence and activity lasting from several seconds to a few minutes. In the present study, simultaneous recordings were made from pairs of TF-related neurons, and the relationships among the firing patterns of cells within a class and between cells of different classes were determined. All cells of a given class showed fluctuations in spontaneous discharge that were in phase. On the other hand, there was a striking reciprocity of firing between the two cell classes, such that a decrease in activity of cells of one class was accompanied by an increase in activity of cells of the other class. These observations point to the existence of integrating mechanisms that coordinate the activity of all members of each class of TF-related neurons. Thus, the pattern of activity of any single on- or off-cell provides a useful index of the excitability of all cells of that class. Moreover, because of the highly reciprocal nature of the firing of the two classes, it is possible to infer the current state of both cell populations from the pattern of activity of any single TF-related neuron. PMID- 2547275 TI - Putative nociceptive modulating neurons in the rostral ventromedial medulla of the rat: firing of on- and off-cells is related to nociceptive responsiveness. AB - In the unstimulated, lightly anesthetized rat, both on- and off-cells exhibit alternating periods of silence and activity lasting from several seconds to a few minutes. In the preceding paper, we showed that the active periods of all cells of the same class are always in phase, whereas the firing of cells of different classes is invariably out of phase. Thus, the pattern of firing of any single on- or off-cell provides a useful indication of the excitability of all on- and off cells in the rostral ventromedial medulla (RVM). In this study, we measured the latency of the tail flick response (TF) at set intervals while recording from TF related neurons in RVM, and were able to demonstrate a significant relationship between the spontaneous firing of both on- and off-cells and the latency of the TF response. If noxious heat is applied at a time when an off-cell is spontaneously active (or an on-cell is silent), the TF latency is longer than if the TF trial falls during a period in which the off-cell is silent (or the on cell is active). This correlation between on- and off-cell firing and changes in TF latency is consistent with a nociceptive modulatory role for either or both cell classes. These findings support the hypothesis that off-cells inhibit and on cells facilitate spinal nociceptive transmission and reflexes. PMID- 2547276 TI - Epidemiological studies of nasopharyngeal cancer in the Guangzhou area, China. Preliminary report. AB - In the city of Guangzhou (Canton) and its surroundings nasopharyngeal cancer (NPC) constitutes about 32% of all cancer and is therefore the most common kind of cancer. Three types of etiological factors seem to be involved: 1) a hereditary factor, 2) Epstein-Barr virus, and 3) external factors in food and water. An epidemiological study has been started in the Guangzhou area with interviews of NPC cases, matching controls and mothers of the cases and controls concerning especially food and environmental factors and infections in childhood. The questions put to the controls and cases covered the last seven years and the age of ten, while those to the mothers covered the first three years of life and the age of ten for the controls and cases. Results are now available for 140 cases, 140 controls and the corresponding living mothers. They indicate that salted fish, particularly of Cantonese style, was consumed more by NPC cases than by controls and also had more frequently been given to weaning children in whom NPC later developed. Furthermore, salted vegetables and salted pork were consumed to a greater extent by NPC cases than by controls. Drinking of polluted water in childhood was more common among the NPC cases. Consumption of fresh fruit was more rare among the cases. Most of the NPC cases had a positive VCA-IgA titre. PMID- 2547277 TI - Enhanced parasympathetic nerve activities in experimentally-induced nasal hypersensitivity. AB - In order to verify the role of cholinergic nerves in nasal allergy and nonspecific nasal hypersensitivity we have evaluated the muscarinic acetylcholine receptors and the activity of choline acetyltransferase in nasal mucosa of TDI sensitized guinea-pigs at specific points in time during a pre-determined sensitization period. The density of muscarinic receptors was significantly elevated in animals sensitized for 8 and/or 12 weeks as well as in the group which received 12 weeks of sensitization followed by a 4 week rest period. Choline acetyltransferase activity was also enhanced at 4-, 8- and 12 weeks. These observations provide biochemical evidence of the hyperactivity of cholinergic nerves in nasal allergy which has been suggested by clinical and pharmacological studies, and lead to the speculation that enhanced parasympathetic nerve function is implicated in causing and maintaining hypersensitivity and hypersecretion in nasal allergy. PMID- 2547278 TI - Histamine content and histamine H1 receptor in experimentally-induced nasal hypersensitivity. AB - Quantitative evaluations of histamine and histamine H1 receptor were carried out in nasal mucosa of toluene diisocyanate (TDI) sensitized guinea-pigs at specific points in time during a pre-determined sensitization period. Histamine content is significantly increased at 1 week and definitely elevated after 2 weeks of sensitization. This increase was maintained throughout the sensitization period. It decreased to normal by four weeks post-sensitization. Also the relatively low histamine content immediately after TDI challenge indicated that a certain amount of histamine was released by TDI challenge. On the other hand, the density and affinity of the histamine H1 receptor was not found to be altered during TDI sensitization. These results suggest that enhanced histamine has a role in the development of nasal allergy, whereas the relevance of quantitative-alteration of the H1 receptor to histamine hypersensitivity observed in nasal allergy has not been demonstrated. PMID- 2547279 TI - Functional disturbances of the sympathetic nerve in experimentally-induced nasal hypersensitivity. AB - Norepinephrine, alpha-1 and beta adrenergic receptors in the nasal mucosa of toluene-diisocyanate sensitized guinea-pigs were evaluated quantitatively at specific points in time during a pre-determined sensitization period. At 4, 8 and 12 weeks, the norepinephrine content was reduced. The concentration of alpha-1 adrenergic receptors significantly decreased in 8- and 12-week groups. A lower density of beta adrenergic receptors was shown in the 12-week group. Signs of a return to normalcy were observed by four weeks post-sensitization. These results led us to speculate that excess norepinephrine release, which may be promoted by allergy-inducing histamine, depleted the norepinephrine at the sympathetic nerve terminal and caused a lowering of the concentration of sympathetic receptors. Functional disturbance of the sympathetic nerve was suggested by clinical findings of nonspecific nasal hypersensitivity, congestion and nasal discharge. PMID- 2547280 TI - Modulation of neuronal choline acetyltransferase activity by factors derived from cultures of non-neuronal cells from the CNS. AB - We have found that cholinergic neurons in spinal cord-dorsal root ganglion cultures derived from E12-E13 mouse embryos are sensitive, as measured by changes in choline acetyltransferase activity, to factors secreted by non-neuronal cells derived from the same tissue at an identical developmental stage. Conditioned medium was produced by incubating non-neuronal cultures for 4 days in defined medium. The cholinotrophic activity present in the conditioned medium had a molecular weight of greater than 50,000 as determined by ultrafiltration and bound wheat germ lectin and heparin sepharose. Total RNA isolated from the non neuronal cells, used to produce the conditioned medium, was translated in frog oocytes. Conditioned medium from the injected oocytes was also found to contain cholinotrophic activity. In contrast, the conditioned medium from water-injected oocytes was inactive. The interaction between the cholinotrophic activity in conditioned medium from frog oocytes and known second messengers was also examined. Dibutyryl cyclic AMP produced a concentration-dependent increase in choline acetyltransferase activity. If a maximal effective dose of dibutyryl cyclic AMP was added in conjunction with a maximal effective dose of conditioned medium from oocytes injected with total RNA a nearly additive response was noted. In contrast, the phorbol ester, phorbol 12-myristate 13-acetate, produced a biphasic change in the level of choline acetyltransferase activity; with lower doses stimulating and higher doses inhibiting the enzyme activity. When conditioned medium from oocytes injected with non-neuronal cell RNA was added in conjunction with the phorbol ester a decrease in the physiological response was noted. PMID- 2547282 TI - Ultrasonic foetal abdominal circumference as a means of assessing gestational age in Nigerians. AB - Foetal abdominal circumference values have been established amongst pregnant Nigerian women between 16 weeks and 40 weeks of gestation, using standard ultrasound methods. A linear mathematical model was found to be adequate in describing the relationship between gestational age and abdominal circumference, and by using the regression equation from the data it was possible to predict the gestational age from knowledge of the abdominal circumference and vice versa. The value of this parameter in assessing foetal age and monitoring high risk pregnancies is discussed. PMID- 2547281 TI - Subdural fluid collections following transcortical approach to intra- or paraventricular tumours. AB - Subdural fluid collections appeared in 15 cases (39%) after removal of 38 intra- and paraventricular tumours in the third or lateral ventricle through 18 frontal and 20 parietal transcortical approaches. Transient fluid collections which disappeared within 2 weeks occurred in 6 cases (16%) and persistent ones in 9 cases (24%). Four of the 9 cases (11%) of collections required surgical treatment because of positive clinical signs and symptoms. Two cases had expansive fluid collections and the other two contained subdural haematomas at surgery. The risk factors likely to contribute to a persistent collection were preoperative ventriculomegaly (frontal horn index greater than 0.38) and a frontal transcortical approach. A symptomatic collection should be considered as a potential complication of the transcortical approach to intraventricular tumours and some methods should be devised to prevent it when intra- or paraventricular tumours with ventriculomegaly are removed. PMID- 2547283 TI - Dimensions of personality and smoking behaviour. AB - The study sample comprised 688 students of a Federal College of Education. Out of this population, there were 109 smokers producing a prevalence rate of 20%. The greatest percentage of smokers fell within the age range 20-29 years. Comparison of smokers and non-smokers showed no statistically significant differences in scores on the extraversion and neuroticism scales of the Eysenck personality questionnaire. Smokers, however, scored significantly higher than non-smokers on the psychoticism scale. An association was found between marital status and smoking behaviour. It is concluded that the association between smoking behaviour and some dimensions of personality such as extraversion and neuroticism reported in some studies may be due to a third variable, most likely a social one. PMID- 2547284 TI - Cardiovascular responses to maximal treadmill exercise in healthy adult Nigerians. AB - Maximum treadmill exercise, using the Bruce protocol was performed on 124 healthy Nigerians (80 males and 44 females) aged 20-59 years. The mean duration of exercise (DOE) and estimated maximal oxygen uptake (VO2 max) decreased linearly with increase in age with a high correlation between age and VO2 max (r = -0.82). Males and active subjects had significantly higher values than females and sedentary subjects respectively. From this study, average values of VO2 max in healthy Nigerian men and women can be predicted from the following regression equations for VO2 max (ml of O2 per kg of body weight per min): Sedentary men = 57.2 - 0.528 (years of age) (r = -0.85), Active men = 65.0 - 0.579 (years of age) (r = -0.82), Sedentary women = 34.6 - 0.236 (years of age) (r = -0.61). The regression equation for active women is not predictive as none above the age of 29 years volunteered to participate in this study. The mean maximal heart rate (MHR) was inversely related to age (r = -0.51) and average value of MHR (beats/min) expected in healthy Nigerian men and women can be predicted from the regression equation below: MHR = 207 - 0.620 (age in years) (r = -0.51). This study presents reference data for the assessment of cardiovascular fitness of Nigerians with or without cardiovascular disease. PMID- 2547285 TI - Hysteroscopy in infertile Nigerian women. AB - The uterine cavity was evaluated by hysteroscopy in 20 Nigerian women aged 23-40 years, as part of the investigations of primary or secondary infertility. Intra uterine abnormalities were detected in 45% of the patients. These abnormalities included intra-uterine adhesions (20%), endometrial polyps (10%), sub-mucous fibroids (5%), endometrial atrophy (5%), and endocervical cysts (5%). This preliminary report reveals a high diagnostic yield with hysteroscopy. The advantages and complications of the procedure are discussed. PMID- 2547286 TI - Mechanisms of enhanced pruritogenicity of chloroquine among patients with malaria: a review. AB - The mechanisms whereby the intrinsic pruritogenic effect of chloroquine (a property also encountered among some other 4-amino-quinolines including amodiaquine) becomes aggravated during paroxysmal malarial suppressive chemotherapy with the drug form the basis of this paper. Physiological itching has been linked to the concept of 'spontaneous itch', as compared to pathological itching which has been associated with another concept of 'itching hyperexcitability', and the pathophysiology of pruritus, including the involvement of peripheral and central (neuropeptide) mediators of itch, were considered. The modulating function of spinal and supraspinal 'gateway control' mechanisms, which have been used to explain the overriding effect of pain-over itch sensation, were also considered and related to itching hyperexcitability. From current data and the records of previously-published reactions to chloroquine, during fever or malarial chemotherapy in man and some mammals, the possible involvement of racial and skin pigment factors, histamine factor, other peripheral mediators of itch, tissue pharmacokinetic factors, central mediators of itch, pyrogenic haemodynamics, and 'gateway' modulation in producing enhanced pruritic reaction during chloroquine antimalarial chemotherapy, were examined in relation to the aggravating role of ischaemia on itch excitability. A trilateral approach to the clinical management of chloroquine-induced pruritus among patients with malaria has been used. In line with the principles of clinical treatment of severe generalized pruritus of uncertain aetiology, this approach has been adapted to reflect the epidemiological, clinical and pathophysiological variables that appear to influence chloroquine-induced pruritus. PMID- 2547287 TI - Diseases causing chronic renal failure in Nigerians--a prospective study of 100 cases. AB - There are indications that there is an increased risk of chronic renal failure (CRF) in the Negroid race, yet few studies have been carried out in the native 'black' environment. A clinico-pathological study of 100 consecutive Nigerian subjects with CRF, seen over a 3-year period, is presented. Primary chronic glomerulonephritis (CGN) accounted for 50, accelerated hypertension for 25, and various aetiological entities for a further nine; these included, chronic pyelonephritis (two), diabetic nephropathy (two), calculous nephropathy (one), toxaemia of pregnancy (one), renal dysplasia (one), tuberculosis (one) and polycystic disease in the ninth subject. In 16 cases, no definitive aetiological diagnosis could be made. Combinations of the following features, protracted hypertension, proteinuria, significant analgesic intake and gouty arthritis, were observed. CGN and accelerated hypertension still remain the leading causes of CRF, while diseases such as diabetes mellitus and chronic pyelonephritis do not contribute significantly to CRF in Nigerians. Recognition of the early features and the causes of CRF would considerably reduce the prevalence of this condition. PMID- 2547288 TI - Serum immunoglobulin levels in the course of normal gestation in Nigerian women. AB - The three major immunoglobulin (IgG, IgA and IgM) concentrations were measured in 118 Nigerian women during normal pregnancy. IgG was found to decline progressively throughout gestation with a very highly significant decrease between the first and second trimester (0.01 greater than P greater than 0.005) and between the second and third trimesters (P = 0.01). The mean concentrations of IgM were significantly lower in sera from first-trimester pregnant women than those of age-matched non-pregnant control subjects (P less than 0.001), and there was a further significant decrease between second and third trimester groups (0.005 greater than P greater than 0.001). No statistical difference was observed for IgA levels throughout normal pregnancy. The significance of these results is discussed in relation to other observations elsewhere. PMID- 2547289 TI - Fine needle aspiration cytology in the assessment of breast lumps in Ibadan. AB - Within the period August 1984 to January 1987, 209 needle aspirates of breast lumps were examined cytologically. One hundred and forty-four smears had histological confirmation. The diagnostic accuracy of fine needle aspiration cytology for malignancy is 79% in this study, with a specificity of 97%. The role of fine needle aspiration cytology, as a useful diagnostic tool and a necessary adjunct to clinical examination in the assessment of breast lumps is discussed. PMID- 2547290 TI - Flexible fibre optic bronchoscopy in Ibadan: report of 6 years' experience. AB - This study describes 6 years' experience of flexible fibre optic bronchoscopy (FFB) at the University College Hospital, Ibadan. Ninety-three bronchoscopies were carried out in 83 patients. The study revealed 22 patients with primary lung malignancies, four with tuberculosis, three with secondary malignancies, three with sarcoidosis, and two with interstitial fibrosis. The yield from FFB was 44% and complications were encountered in only three patients, with no mortality. PMID- 2547291 TI - The levels of zinc in breast milk of urban African women in Zimbabwe. AB - Two hundred and eighty milk samples expressed by healthy African women who had pre-term or full-term delivery, were digested using the wet digestion method. The digests were analysed for zinc (Zn) using an atomic absorption spectrophotometer. Mean Zn level in colostrum was 8.2 +/- 1.02 micrograms/ml in transitional milk 4.33 +/- 0.134 micrograms/ml and in mature milk 0.38 +/- 0.03 micrograms/ml. The ranges were 1.4-21.7, 0.3-9.8 and 0.2-2.4 micrograms/ml in colostrum, transitional and mature milk respectively. The figures obtained agreed with some previous findings in which it was found that Zn levels decreased with progression in lactation. PMID- 2547292 TI - Echocardiographic features of mitral valve prolapse in Libyan patients. AB - Echocardiographic observations in 200 subjects with mitral valve prolapse (MVP) are presented. The diagnostic criteria used were: (1) abrupt late systolic posterior motion of one or both leaflets of the mitral valve, and (2) holoor pansystolic posterior motion of 3 mm of one or both leaflets of the mitral valve. Most of the subjects were young--72% were aged less than 30 years. Prolapse of posterior leaflets was noted in 98% of subjects--69.5% late systolic, 28.5% pansystolic, and 2% had prolapse of the anterior mitral leaflet only. Mitral valve prolapse was considered to be primary--being the only abnormality in 78.5% of the subjects. In the remaining 21.5% MVP was associated with other cardiac lesions, the commonest being, atrial septal defect (2.5%), dilated aortic root (2%), bicuspid aortic valve (2%), cardiomyopathy (5%), rheumatic heart disease (4%) and ischaemic heart disease (1.5%). Mitral valve prolapse was considered to be important enough to result in haemodynamically significant mitral regurgitation in only 8% of subjects. Mitral valve prolapse was the commonest single echocardiographic abnormality (16%) observed in patients referred to this university hospital, which is the referral centre for approximately half of Libya. Although this does not indicate the prevalence of MVP in the general population, this study indicates MVP to be the commonest valvular abnormality seen in hospital practice in Libya. PMID- 2547293 TI - Comparison of the partitioning in vitro of chloroquine and its desethyl metabolites between the erythrocytes and plasma of healthy subjects and those with falciparum malaria. AB - The partitioning of chloroquine and its two desethyl metabolites between red blood cells (RBCs) and plasma was studied in vitro, using blood from healthy adults and from children with Plasmodium falciparum parasitaemia. Blood from the volunteers was incubated with varying concentrations of chloroquine (CQ), desethylchloroquine (DCQ) and bisdesethyl-chloroquine (BDCQ) for 15 min and the RBC/plasma concentration ratio determined. Desethylchloroquine and BDCQ were concentrated in the red cells of uninfected blood to the same extent as chloroquine. On the other hand, DCQ and BDCQ were concentrated to a significantly lower extent than CQ in the red cells from malarial children. The reduced ability of infected RBCs to concentrate DCQ and BDCQ may have an important bearing on the development of resistance to chloroquine by P. falciparum. PMID- 2547294 TI - Nutritional counseling. U.S. Preventive Services Task Force. AB - Clinicians should provide periodic counseling regarding dietary intake of calories, fat (especially saturated fat), cholesterol, complex carbohydrates (starches), fiber and sodium. Women and adolescent girls should receive counseling on calcium and iron intake, and pregnant women should receive specific information on nutritional guidelines during pregnancy. Patients should also be counseled about the nutritional requirements of infancy and early childhood. PMID- 2547295 TI - Fibrolamellar hepatocellular carcinoma. AB - Fibrolamellar hepatocellular carcinoma is a primary malignant tumor of the liver that usually affects adolescents and young adults. If detected early, this cancer is frequently resectable, with a much longer survival time than in classic hepatocellular carcinoma. Fibrolamellar hepatocellular carcinoma has distinct radiographic and pathologic features. Serum unsaturated vitamin B12 binding capacity and plasma neurotensin may be used as tumor markers. PMID- 2547296 TI - Lack of evidence for alpha-adrenergic receptor-mediated mechanisms in the genesis of ischemia in syndrome X. AB - Patients with syndrome X (typical angina pectoris, positive exercise tests [greater than or equal to 1 mm of ST-segment depression], no evidence of coronary spasm and angiographically normal coronary arteries) have a reduced coronary flow reserve due to inappropriate dilatation of small resistive vessels. To assess whether alpha-adrenergic mechanisms play a role in the genesis of ST-ischemic changes in syndrome X, 12 patients with this syndrome (2 men and 10 women, mean age 50 +/- 6 years) underwent exercise testing and 24-hour ambulatory electrocardiographic monitoring. They were done off treatment and after alpha blockade with prazosin and clonidine on 2 separate weeks. Despite treatment, all exercise tests remained positive and patients were stopped because of progressive angina pain. Compared to the off-treatment tests, exercise duration and heart rate-blood pressure product at 1 mm of ST-segment depression did not change significantly after prazosin (617 +/- 203 vs 663 +/- 203 seconds and 23,857 +/- 6,125 vs 22,098 +/- 4,816 beats/min X mm Hg, respectively) and clonidine (684 +/- 148 vs 649 +/- 80 seconds and 25,514 +/- 2,386 vs 24,567 +/- 2,001 beats/min X mm Hg, respectively). Ambulatory monitoring showed similar results regarding number of episodes of ST-segment depression greater than or equal to 0.1 mV during control and after prazosin (39 vs 38) or clonidine (26 vs 23) treatment. None of the 8 patients who also underwent provocative testing with phenylephrine had ischemic electrocardiographic changes; only 2 experienced chest pain during the test.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547297 TI - Painful versus silent myocardial ischemia during leg and arm exercise testing in stable angina pectoris. AB - When 51 patients with proven coronary heart disease and stable angina pectoris underwent exercise testing, 22 experienced painful myocardial ischemia during both leg and arm exercise testing (group L + A), whereas 29 patients had such episodes only during the leg testing (group L). Upright bicycle exercise was performed with the legs first, followed 2 days later by arm testing. Exercise was stopped when typical anginal pain and greater than 1-mm ST horizontal depression occurred during leg testing, and when greater than 1-mm ST horizontal depression was noted during arm testing. Heart rate, systolic blood pressure and rate pressure product for leg and arm testing, either at the beginning of anginal pain or at the time when 1-mm ST depression was noted, were similar. Two-dimensional echocardiography showed that the L group had higher (p less than 0.01) end systolic volume at rest and decreased (p less than 0.05) ejection fraction during exercise. Coronary angiography showed that the L group had a greater (p less than 0.001) number of patients with 3-vessel disease, a decreased (p less than 0.001) ejection fraction and less patients with 1-vessel disease. In these patients, absence of anginal pain during arm exercise suggests defective segmental transmission of pain sensation related to severe coronary artery disease. Thus, arm testing, in addition to leg testing, seems to be a simple and useful tool for the detection of severe coronary disease. PMID- 2547298 TI - Association of coronary artery disease in cardiac transplant recipients with cytomegalovirus infection. AB - Coronary artery disease (CAD) is now the major limitation to long-term survival after cardiac transplantation. Its etiology remains unclear. The possible role of viral infection in the genesis of CAD stimulated the review of 102 patients transplanted since the introduction of triple drug immunosuppression (cyclosporine, azathioprine and prednisone) to assess the importance of posttransplant cytomegalovirus infection in the development of CAD in the cardiac graft. CAD occurred in 16 patients (16%). Recipient age and sex, donor age, pretransplant diagnosis, frequency of acute rejection episodes, HLA mismatch, cytomegalovirus infection, incidence of posttransplant systemic hypertension and diabetes mellitus, and mean triglyceride, cholesterol and cyclosporine levels were analyzed to assess their influence on the development of CAD. Only the occurrence of cytomegalovirus infection was found to be a significant factor (p = 0.007): infection occurred in 62% of patients with CAD and in only 25% of those without. These data support the existence of an association between cytomegalovirus infection and CAD after cardiac transplant. It is possible that the virus contributes to the initial injury to the coronary endothelium. PMID- 2547299 TI - Interactions between circulating peptides and the central nervous system in hemodynamic regulation. AB - Enkephalins and endothelins are endogenous peptides, which, at least at pharmacologic doses, produce complex hemodynamic responses after intravenous administration. The enkephalins, when injected into conscious animal models and humans, increase blood pressure, heart rate and minute ventilation. This response occurs by activation of specific opiate receptors located outside the bloodbrain barrier; the actual mechanism involves an increase in adrenergic autonomic nervous system tone and a decrease in cholinergic tone. These opiate receptors may activate afferent fibers, perhaps nicotinic cholinoceptors; in many ways their properties are suggestive of chemoreceptors. Furthermore, enkephalin responses appear to be modulated by gamma-aminobutyric acid complexes, in that the reversal of the excitatory hemodynamic responses seen in the conscious state to vasodepressor responses after barbiturate anesthesia may result from alteration of the state of activation of the gamma-aminobutyric acid complex. The enkephalin receptors are localized to the vertebral artery vascular distribution; the specific site may be the area postrema, a blood-brain barrier-deficient circum-ventricular organ demonstrated to modulate heart rate and blood pressure and to represent a target site for circulating angiotensin II. Endothelin increases heart rate and blood pressure when infused slowly into conscious or anesthetized dogs, although barbiturates do blunt the increase in heart rate. The mechanism appears to involve modification of autonomic tone, but also some element of direct vasoconstrictor activity. Interestingly, rapid bolus doses of endothelin produce only vasodepressor responses, suggesting that the rate and concentration at which circulating endothelin reaches afferent receptors or vasoconstrictor sites on vascular smooth muscle may determine the net hemodynamic response observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547300 TI - Blood glucose response to pea fiber: comparisons with sugar beet fiber and wheat bran. AB - Two new fiber types, pea fiber (PF) and sugar beet fiber (BF), were compared with wheat bran (WB) to investigate the effect on postprandial blood glucose and serum insulin responses in normal subjects. The control meal consisted of 150 g ground beef mixed with 50 g glucose and 20 g lactulose. Only addition of PF (15 g pure fiber) reduced the area under the incremental blood glucose curve significantly (by 65%, p less than 0.05). None of the fibers affected the area under the insulin-response curve significantly although it was reduced by all fibers. Mouth to-cecum transit time, assessed by the hydrogen breath technique, was decreased by WB and BF, (p less than 0.05) but not by PF. PF is palatable and may prove beneficial as a fiber supplement for diabetics. PMID- 2547301 TI - Long-term nutritional status of an enterally nourished institutionalized population. AB - Growth and nitrogen nutriture were evaluated during a yearlong study of 11 profoundly retarded nonambulatory institutionalized youth who were fed by gastrostomy and received anticonvulsants. The effects of dietary fiber and vitamin D were assessed by determining plasma levels and balances of zinc, magnesium, calcium, and phosphorus in a subset of six patients. Increase in body weight was 3.4 +/- 2.4 kg (means + SD), in height, 5.9 +/- 3.9 cm during the year. N balance remained positive throughout the study. Zn and Ca balances suggested that needs for this population may be greater than or equal to 150% of the Recommended Dietary Allowance. Vitamin D supplementation had no effect. Soy polysaccharide fiber intakes of 18-24 g/d compared with 0 or 12-20 g fiber/d significantly improved N and P retentions. Although the determination of nutrient needs of a severely disabled population is complex, our results suggest N and energy but not Zn and Ca allowances for a healthy population may be satisfactory guidelines. PMID- 2547302 TI - Neutron radiation therapy for unresectable non-small-cell carcinoma of the lung. A review. AB - Over 200 patients have been entered in five studies investigating the use of fast neutron radiation therapy in the treatment of non-small-cell carcinomas of the lung since 1983. The results of these studies have been inconsistent. Most studies did not show survival rates or local control advantages over standard photon radiation therapy. Side effects from studies employing mixed photon neutron treatment plans or clinically oriented, high-energy cyclotrons were seen to be comparable to those of standard courses of radiation therapy, representing a considerable improvement over those studies utilizing low-energy cyclotrons for a full course of radiation therapy, which resulted in unacceptably high complication rates. A new phase III study utilizing high-energy isocentric neutron beams has been designed and implemented, and over 100 patients have been entered to date. The current status of fast neutron radiation therapy in the treatment of non-small-cell lung cancer is reviewed. PMID- 2547303 TI - Neutron radiotherapy for malignant gliomas. AB - Neutron radiotherapy has been used for patients with malignant gliomas for over a decade; a substantial number of patients have been treated to date. Pathologic analysis of surgical specimens posttreatment and autopsy specimens have documented an increased antitumor effect of neutrons against malignant gliomas, compared with photon irradiation. However, results of neutron trials to date have not shown a survival advantage over conventional radiotherapy for these patients. This article reviews current surgical, radiotherapeutic, and chemotherapeutic approaches to these tumors, the rationale for neutron treatment, and the results of trials of neutron radiotherapy conducted to date for patients with malignant gliomas. PMID- 2547305 TI - Chemotherapy of advanced non-small-cell lung cancer: a randomized trial of three cis-platin-based chemotherapy regimens. AB - One hundred fifty-two patients with locally advanced or metastatic non-small-cell lung cancer were randomized to receive treatment with one of three cis-platin containing chemotherapy regimens: vindesine/cis-platin, etoposide/cis-platin, or vindesine/etoposide/cis-platin. Following an 8-week induction course of treatment, patients were evaluated for response; responders continued to receive monthly chemotherapy. All patients were followed until death. Response rates for the three regimens were 10%, 6%, and 24%, respectively; the cis platin/vindesine/etoposide regimen produced more responses than did either cis platin/etoposide or cis-platin/vindesine (p less than 0.05). However, median survival was not improved with the three-drug regimen, and myelosuppression produced by this regimen was worse. The 20-week median survival for the entire group suggests that these treatments had no impact on survival. None of these regimens can be recommended for routine treatment of patients with advanced non small-cell lung cancer. The addition of vindesine did not make a significant impact in response rate or overall survival in this study. PMID- 2547304 TI - Alternating chemotherapy with or without VP-16 in extensive-stage small-cell lung cancer. AB - In this study, we evaluated the role of alternating chemotherapy with or without etoposide (VP-16) in patients with extensive-stage small-cell carcinoma (SCC) of the lung. All patients received initial treatment with CMC [cyclophosphamide, methotrexate, and chloroethyl-cyclohexyl-nitrosourea (CCNU)]. Four weeks after initial treatment, patients were stratified by performance score, central nervous system (CNS) metastasis, age, and response to initial CMC therapy and randomized to receive AO (doxorubicin and vincristine) or AVO (doxorubicin, VP-16, and vincristine) alternating with CMC. One hundred eighty-two eligible patients were treated with the initial cycle of CMC and 98 responded (54%). One hundred fifty four patients were randomized to either AO/CMC or AVO/CMC. The response rates to AO/CMC and AVO/CMC were similar (72 vs. 68%). The time to progression and survival were not significantly different on the two treatment regimens. Toxicity was significantly greater for patients receiving AVO/CMC with six treatment related deaths. Etoposide as used in this regimen did not significantly influence response rates, time to progression, or survival of patients with extensive small cell lung cancer. PMID- 2547306 TI - Phase II study of iproplatin (CHIP) in previously treated small-cell lung cancer. AB - Eighteen patients with previously treated extensive small-cell carcinoma of the lung were entered into a Phase II study employing iproplatin (CHIP), a cis-platin analog. Patients had received a mean of two prior chemotherapeutic regimens. Fifty-five percent had received prior cis-platinum and 33% had received prior radiation therapy. CHIP (225 mg/m2) was administered by intermittent intravenous infusion over 30 min for 5 days of a 28-day cycle without prehydration. Sixteen patients with Zubrod performance scores of less than or equal to 3 received 27 courses of therapy (mean 1.7, range 1-6). One partial response of 167 days duration was observed, with complete regression of involved lymph nodes and stabilization of nonevaluable disease in the chest. Six patients had stable disease following one cycle of CHIP, but all progressed following a second cycle of drug. The main toxicity was myelosuppression with prominent thrombocytopenia. Median survival was 75 days from initiation of therapy. In this group of heavily pretreated patients with advanced disease, iproplatin has only minimal activity as a single agent and does not show non-cross-resistance with cis-platinum. PMID- 2547307 TI - Immunohistochemical study of lung adenocarcinoma using monoclonal antibody for 60 kilodalton antigen in type II pneumocytes and nonciliated bronchiolar epithelial cells. Comparison with two antisurfactant apoprotein antibodies. AB - Monoclonal antibody KP16D3 was produced by immunizing mice with monkey bronchoalveolar lavage. KP16D3 revealed the immunohistochemical reactivity in the cytoplasm of some nonciliated bronchiolar epithelial cells and type II pneumocytes and thereby recognized specifically a protein with an apparent molecular weight of 60 kD with the use of Western blotting and immunoaffinity column chromatography followed by SDS-PAGE. Examination of 76 primary and 4 metastatic lung carcinomas in primary lung carcinoma KP16D3 showed immunohistochemical positivity only to mucin-nonproducing papillary adenocarcinoma (27/28) and bronchioloalveolar carcinoma (2/2), except for one case of large cell carcinoma. All other primary lung carcinomas such as squamous cell carcinoma, acinar adenocarcinoma, and small cell carcinoma had negative results. From these findings, KP16D3 seems to be an effective immunohistochemical marker of mucin-nonproducing papillary adenocarcinoma and bronchioloalveolar carcinoma of the lung and it appears to be useful to investigate both the histogenesis and functional expression of primary lung adenocarcinoma. PMID- 2547308 TI - The clinical spectrum, pathology, and clonal analysis of Epstein-Barr virus associated lymphoproliferative disorders in heart-lung transplant recipients. AB - This study presents the clinical and laboratory observations on posttransplant lymphoproliferative disorders (PTLDs) occurring in 5 of 53 heart-lung transplantation recipients. Cervical lymph nodes, tonsils, lungs, and gastrointestinal tract were the common sites of involvement by PTLDs. The histopathologic findings showed a spectrum of lymphoid and immunoblastic proliferation ranging from diffuse hyperplasia to malignant lymphoma, immunoblastic or large cell type. All cases were associated with a primary Epstein-Barr virus infection, and viral DNA was demonstrated within the lesional tissue in three cases. Immunohistochemical and immunoglobulin gene rearrangement studies revealed a B-cell proliferation that was monoclonal in three cases and polyclonal in two cases. Compared with PTLDs arising in other organ transplant recipients, this series is remarkable for a high incidence of PTLDs (9.4%), a short interval to tumor diagnosis (2.2 months, mean), involvement of the primary allograft in three cases (60%), and the frequent development of bronchiolitis obliterans. Possible reasons for this distinct clinicopathologic profile are discussed. PMID- 2547310 TI - Latex kits for detection of rotavirus. PMID- 2547309 TI - Plasma cell malignancy in the acquired immune deficiency syndrome. Association with Epstein-Barr virus. AB - The development of high-grade, malignant B-cell lymphoma is a well-recognized complication of human immunodeficiency virus (HIV) infection. Plasma cell neoplasms, however, have been rarely encountered in HIV-infected people. This study presents the morphologic and immunologic features of an unusual plasma cell tumor occurring in a 31-year-old HIV-antibody-positive male. The malignancy was characterized by widespread dissemination and hypercalcemia at presentation and a clinically aggressive course. Immunoperoxidase staining of tumor tissue obtained from biopsy and at autopsy had positive results for IgM and lambda. In the patient's serum, only an IgG kappa paraprotein was detected, indicating that the tumor was nonsecretory. DNA analysis of autopsy-derived tumor tissues demonstrated clonal rearrangements of the immunoglobulin (Ig) heavy chain gene locus and rearrangements in both kappa and lambda light chain gene loci. Furthermore, DNA hybridization studies revealed the presence of Epstein-Barr virus (EBV) genomes in tumor tissue but not in nontumor tissue from this patient. PMID- 2547311 TI - EBV-DNA in lymphoid tissue. PMID- 2547312 TI - Mouth-to-cecum transit time in patients affected by chronic constipation: effect of glucomannan. AB - Mouth-to-cecum transit time was studied in 13 patients affected by chronic idiopathic constipation and 18 control subjects matched with the constipation group for age, sex, and dietary habits. In a preliminary investigation, all patients showed a prolonged whole gut (oroanal) transit time as measured with radiopaque markers. Mouth-to-cecum transit time was studied through the serial determination of breath H2 after administration of 12 g lactulose diluted in 120 ml water. Breath H2 was measured with a gas analyzer and was determined in parts per million (ppm). Breath H2 after lactulose was also determined in the group with constipation after a 10-day diet that included either glucomannan (1 g tid orally) or placebo administered in a double-blind manner. The results show a statistically significant increase in mouth-to-cecum transit time in the group with constipation, compared with controls, and a return to within the normal range after the 10-day treatment with glucomannan. With placebo, no difference in transit time was noted. We therefore suggest that chronic idiopathic constipation is a disease that involves the whole gut. PMID- 2547313 TI - Hereditary adenomatosis of the colon and rectum: clinical features of eight families from northern Italy. AB - Adenomatosis coli (or familial polyposis of the large bowel) and related syndromes are relatively rare diseases characterized by an autosomal dominant mode of inheritance. In these diseases, the entire colorectal mucosa is covered by hundreds (often innumerable) polyps of various dimensions. In addition, several extracolonic abnormalities have been reported. In the present study, we describe the clinical features of eight families from northern Italy fulfilling the diagnostic criteria of adenomatosis coli. Information was available on 123 unaffected and 30 affected family members. The most relevant findings of the study can be summarized as follows. 1) Gene frequency was calculated to be between 1:7,300 and 1:19,000. Segregation ratio in affected branches was 0.57, with a gene penetrance of nearly 60% and a male:female ratio of 1.73. 2) Extracolonic manifestations were present in all families and in 15 of 30 affected patients, the most frequent being cutaneous cysts and retinal lesions. No case fulfilling the classical criteria of Gardner syndrome was observed. 3) When the diagnosis of adenomatosis followed the appearance of symptoms, colorectal cancer had usually already developed, whereas no malignant changes were observed in individuals diagnosed in the asymptomatic stage. When colectomy with ileorectal anastomosis was the treatment of choice, polyps tended to recur in the rectal stump, and long-term endoscopic follow-up was necessary. In conclusion, adenomatosis coli may account for a definite proportion of colorectal neoplasms observed in the general population. Taking into consideration the genetic base of the disease, it follows that individuals at risk should be closely monitored for several years. Moreover, clinical investigations should not be limited to the large bowel, but should be extended to the skin, upper digestive tract, fundus oculi, bones, and probably other organs. PMID- 2547315 TI - Epsilon-aminocaproic acid in the reversal of consumptive coagulopathy with platelet sequestration in a vascular malformation of Klippel-Trenaunay syndrome. PMID- 2547314 TI - Inhibition of adenylate kinase by P1,P5-di(adenosine 5') pentaphosphate in assays of erythrocyte enzyme activities requiring adenine nucleotides. AB - P1,P5-di(adenosine 5')pentaphosphate (Ap5A) is an excellent inhibitor of human hemolysate adenylate kinase at concentrations near 2 microM and above. At ten times this concentration and in hemolysate enzyme assays under conditions described in this paper it appears not to alter reaction data in the case of hexokinase, phosphofructokinase, and phosphoglycerokinase. In the pyruvate kinase assay, very modest reductions in activity are noted, and kinetics with phosphoenolpyruvate, adenosine diphosphate (ADP), and uridine diphosphate (UDP) are unaltered. PMID- 2547316 TI - Aldosterone response to insulin-induced hypoglycemia in hemodialysis patients. AB - To investigate possible abnormalities of the hypothalamic-pituitary axis in patients with chronic renal failure on dialysis, we have examined the effects of insulin-induced hypoglycemia on the adrenal steroid responses. In normal subjects, plasma aldosterone and cortisol concentrations increase significantly in response to hypoglycemia, with good correlation. In the patients with end stage renal disease (ESRD) however, insulin-induced hypoglycemia fails to elicit significant increases in the plasma cortisol and aldosterone levels. To test the adrenal responsiveness to adrenocorticotropin (ACTH), we administered ACTH to both groups. Plasma cortisol and aldosterone responses are similar in both groups suggesting that the adrenal responsiveness to ACTH is not impaired. We also investigated the responsiveness of the renin-angiotensin-aldosterone system in response to volume contraction by hemofiltration in patients with ESRD. Neither plasma renin activity nor plasma aldosterone concentration change significantly following such contrived volume contraction. These results reveal several endocrinologic abnormalities in the patients with ESRD on chronic hemodialysis. PMID- 2547317 TI - Topography of early HPV 16 transcription in high-grade genital precancers. AB - The extent to which human papillomavirus (HPV) type 16 is transcribed and the nature of the transcripts produced in genital precancers has not been clearly defined. The authors analyzed 28 cases of cervical (CIN) or vulvar (VIN) intraepithelial neoplasia by RNA-RNA in situ hybridization, using probes generated from HPV 16 open reading frames (ORFs) either upstream (E6-E7) or downstream (E2-E5-L2) to the E1 ORF, where HPV 16 genomic integration most commonly occurs. Hybridization signals corresponding to one or both probes were detected in a high proportion of cells throughout the lesional epithelium of low- and high-grade CIN, including basal layers. In serial sections analyzed with the two probes, hybridization signals were obtained from both, and in similar proportion, irrespective of CIN grade. The distribution and character of hybridization signals suggests that the morphologic progression of precancers is not associated with either cessation of HPV 16 early transcription or a change in the general character of the transcripts produced. PMID- 2547318 TI - Alterations in immunoglobulin genes reveal the origin and evolution of monotypic and bitypic B cell lymphomas. AB - During progression of B-lymphoproliferative disorders (B-LPD), increasing divergence can be detected in histology, cytogenetics, and clinical behavior. To investigate genomic tumor cell heterogeneity, 50 biopsies of 21 patients with B LPD of different histogenetic origin were studied for changes in the immunoglobulin gene structure during follow-up study. Ig-heavy chain (IgH) gene alterations were analyzed by Southern blotting using a panel of eight endonucleases. Ig-light chain (IgL) genes and the translocation of the bc 1-2 gene involved in t(14;18) were also studied. In seven of nine follicular lymphomas, most alterations suggested mutations in the IgH genes. Conservation of most restriction sites and of the t(14; 18) breakpoint confirmed the monoclonal origin in these tumors. Also, in two of three diffuse follicle center cell lymphomas (CLL) and in each of four immunocytomas, IgH alterations were found. In contrast, clonal changes were absent in three centrocytic lymphomas and two cases of CLL. In two follicular lymphomas with bitypic IgL expression, a common origin with subsequent divergence of the two constituents, rather than true biclonality, was indicated by the Ig gene structures. No relation was found between the frequency of somatic mutations and histologic signs of progression. These data indicate that somatic hypermutation in IgH genes is related to the histogenesis of the B-LPD and reflect the physiology of their benign counterparts in normal B cell development. PMID- 2547319 TI - Biphasic cellular and tissue response of rat lungs after eight-day aerosol exposure to the silicon dioxide cristobalite. AB - Cristobalite is a crystalline silicon dioxide that elicits pulmonary inflammation and fibrosis in humans and experimental animals. Exposure of rats to aerosols of respirable cristobalite for 8 days led to a rapid influx of neutrophils and macrophages into alveolar and tissue compartments of the lung followed by a more gradual accumulation of T lymphocytes. This inflammatory response persisted throughout 52 weeks after the end of the exposure. For some variables studied there appeared to be a cyclical nature to the response. Statistical analysis of alveolar cell populations and lung tissue weight, protein, and hydroxyproline showed significant time-dependent fluctuations. Histologic analysis revealed a progressive deposition of collagen and type II cell hyperplasia centered on airways, however, there appeared to be some correlation between fluctuations in alveolar cell populations and overall tissue pathology. The observed cellular and biochemical fluctuations and the persistence of the inflammatory response may be due to the presence of silica in the lung, which serves as a source of repetitive stimulation of lung cells. PMID- 2547321 TI - A high-performance liquid chromatographic method for the determination of polyphosphoinositides in brain. AB - A simple high-performance liquid chromatographic method for the determination of endogenous phosphatidylinositol 4-phosphate (PIP) and phosphatidylinositol 4,5 bisphosphate (PIP2) in brain has been developed. PIP and PIP2 were derivatized with 9-anthryldiazomethane to yield (9-anthryl)PIP and di(9-anthryl)-PIP2. The derivatives were separated on a reversed-phase column using isocratic elution and detected with a uv detector. The detection limits of PIP and PIP2 were 0.25 micrograms. The method with uv detection was sufficiently sensitive to measure the concentrations of PIP and PIP2 in rat brain. The levels of PIP and PIP2 were increased in developing rat brain and were decreased after 10 min of ischemia. PMID- 2547320 TI - Colocalization of elastase and myeloperoxidase in human blood and bone marrow neutrophils using a monoclonal antibody and immunogold. AB - The authors have localized elastase in human blood and bone marrow neutrophils by immunoelectron microscopy using a monoclonal anti-human elastase antibody (NP 57) and compared its distribution with myeloperoxidase (MPO) and lactoferrin (LF), which mark primary and secondary neutrophil granule, respectively. Human bone marrow and blood polymorphonuclear leukocytes (PMN), either unstimulated or after phagocytosis of latex microbeads, were fixed in 4% paraformaldehyde. Ultrathin frozen sections were immunolabeled with NP 57, followed by an immunogold probe. In bone marrow granulocyte precursors elastase appeared simultaneously in the immature first granules of myeloblasts with MPO. As these granules became denser with maturation, labeling for both enzymes became weaker and sometimes negative (possibly due to masking of immunoreactivity). The ellipsoidal primary granules were strongly labeled by NP57. LF positive granules appeared later, at the myelocyte stage, and contained neither MPO nor elastase. In mature neutrophils, immunolabeling for elastase was found together with MPO in the large electron dense primary granules and in a different granule population from the LF-positive secondary granules. Double labeling with two different-sized gold particles was used to compare the kinetics of degranulation of secondary and primary granules. The observation and the analysis of single phagosome content was made possible by this new technique. In conclusion, immunoelectron microscopy was used to show elastase in the primary granules of neutrophils, where it appears simultaneously with MPO. This technique has also allowed comparison of the kinetics of degranulation of both types of granules, and could be applied to different experimental and pathologic conditions. PMID- 2547322 TI - High-performance liquid affinity chromatography with phenylboronic acid, benzamidine, tri-L-alanine, and concanavalin A immobilized on 3 isothiocyanatopropyltriethoxysilane-activated nonporous monodisperse silicas. AB - Nonporous, microparticulate, monodisperse silicas with particle diameters between 0.7 and 2.1 microns are introduced as stationary phases in high-performance affinity chromatography. The immobilization of m-aminophenylboronic acid, p aminobenzamidine, tri-L-alanine, and concanavalin A onto these silicas was successfully achieved using 3-isothiocyanatopropyl-triethoxysilane as an activation reagent. Immobilized phenylboronic acid was applied to the isolation of nucleosides, nucleotides, and glycoprotein hormones such as bovine follicotropin and human chorionic gonadotropin, while immobilized benzamidine was employed for the isolation of the serine proteases thrombin and trypsin, immobilized tri-L-alanine for the separation of pig pancreatic elastase and human leukocyte elastase, and immobilized concanavalin A for the isolation of horseradish peroxidase. In all affinity chromatographic systems studied, the nonporous monodisperse silicas showed improved chromatographic performance compared to results obtained with porous silica supports using identical activation and immobilization procedures. Furthermore, frontal analysis was used as a method to evaluate the influence of experimental parameters on biological activity and accessible ligand densities. Only minor changes in bioactivity were found with the nonporous affinity supports, where accessibilities were typically higher than ca. 60%. The immobilization of affinity ligands onto porous supports as used in this and associated papers thus represents a successful general procedure for the preparation of stable matrices with fast kinetics for use in high-performance affinity chromatography. PMID- 2547323 TI - Application of nonionic polyoxyethylene detergents in studies of phosphatidylinositol kinase. AB - Four commercial nonionic polyoxyethylene detergents were compared with Triton X 100 in terms of physicochemical properties and interaction with an integral membrane protein, phosphatidylinositol kinase. Tergitol 15-S-9, Tergitol TMN-10, polyoxyethylene 10 lauryl ether, and polyoxyethylene 10 tridecyl ether possess aliphatic hydrophobic moieties and therefore do not absorb significantly in the ultraviolet region. Like Triton X-100, these detergents have cloud points well above the physiological temperature range and partial specific volumes greater than 0.9 cm3/g. Critical micelle concentrations range from 0.02 to 0.03% with the exception of Tergitol TMN-10, for which the value is about 0.15%. All five detergents support activity of phosphatidylinositol kinase from rat liver in a mixed micellar assay and show similar ionic strength dependences for solubilization of the enzyme. Enzyme/detergent mixed micelles are somewhat smaller with Triton X-100 than for the other detergents. The data presented suggest that these detergents may be generally substituted for Triton X-100 in applications where uv invisibility is important. PMID- 2547324 TI - An assay for superoxide dismutase activity in mammalian tissue homogenates. AB - During the course of measuring superoxide dismutase (SOD) activity in rat breast tissue, interferences in the nitroblue tetrazolium (NBT) and cytochrome c assay systems were noted. These interferences inhibit accurate measurement of SOD activity in breast tissues, necessitating the development of a new NBT-based assay that includes compounds capable of inhibiting tissue specific interferences. The most effective compounds were metal chelators that were also electron transport chain inhibitors. Bathocuproine sulfonate (BCS) was the most effective of these compounds. The inclusion of BCS in the NBT assay system was shown to make the accurate measurement of SOD activity in tissues with interferences possible. PMID- 2547325 TI - Pipecuronium-induced neuromuscular blockade during nitrous oxide-fentanyl, isoflurane, and halothane anesthesia in adults and children. AB - To determine in adults and children the dose-response relationship and the duration of action of pipecuronium bromide during fentanyl-nitrous oxide (N2O), isoflurane, and halothane anesthesia, the authors studied 30 ASA Physical Status 1-2 adults (age: 16-55 yr) and 30 ASA Physical Status 1-2 children (age: 1.7-11.5 yr) during minor elective surgery. Patients were anesthetized with N2O/O2 (60:40) supplemented with either fentanyl (4 micrograms/kg), or isoflurane (adults, 0.9%; children, 1.2%), or halothane (adults, 0.6%; children, 0.7%). Neuromuscular (NM) blockade was measured by electromyography. Incremental iv doses of pipecuronium were administered to determine the cumulative dose-response relationship of pipecuronium until a 95% twitch depression (ED95) had been obtained. In adults, ED50 was 31.7 +/- 2.9 micrograms/kg (mean +/- SE) during fentanyl-N2O/O2, reduced by isoflurane (18.0 +/- 4.8 micrograms/kg, P less than 0.05) but not by halothane (25.0 +/- 2.6 micrograms/kg, NS). ED95 was 59.4 +/- 5.4 micrograms/kg during fentanyl-N2O/O2, reduced by isoflurane (42.3 +/- 2.5 micrograms/kg, P less than 0.05), but not by halothane (49.7 +/- 3.1 micrograms/kg, NS). In children, ED50 was 43.9 +/- 4.7 micrograms/kg during fentanyl-N2O/O2, reduced by isoflurane (23.1 +/- 1.6 micrograms/kg, P less than 0.05), and halothane (33.2 +/- 3.2 micrograms/kg, P less than 0.05). ED95 was 79.3 +/- 9.8 micrograms/kg during fentanyl-N2O/O2, and reduced by isoflurane (49.1 +/- 3.1 micrograms/kg, P less than 0.05), but not by halothane (62.5 +/- 7.3 micrograms/kg, NS). Comparison between adults and children reveals no statistically significant differences, except for ED50 during fentanyl-N2O/O2 anesthesia which was increased in children.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547326 TI - Inhibition of mobilization of acetylcholine: the weak link in neuromuscular transmission during partial neuromuscular block with d-tubocurarine. AB - The authors have demonstrated earlier, by direct measurement of acetylcholine (ACh), that d-tubocurarine (d-Tc) and other nondepolarizing muscle relaxants decrease the release of ACh from the indirectly stimulated mouse hemidiaphragm preparation. It was the purpose of the present study to determine whether the decrease of the stimulated release of ACh and the increase in the intensity of the partial neuromuscular block observed at higher stimulation rates is caused by the inhibition of mobilization of ACh from reserve depots to release sites or by inhibition of the release process itself. To attain our objective, the authors have investigated the influence of the progressive increase of the rate of stimulation from 0.1 to 1, 2, 3, and 5 Hz on the force of contraction on the in vitro phrenic nerve-hemidiaphragm and in vivo sciatic nerve-tibialis anterior preparation in the absence of drugs and after about 20% neuromuscular block produced by d-Tc or by Mg2+. The latter is known to inhibit the Ca2+ dependent release of ACh. In the absence of drugs increasing the stimulation rate increased the force of contraction, in vivo and in vitro, during both indirect and direct stimulation. In the phrenic nerve-hemidiaphragm preparation the increase was significant at 1, 2, 3, and 5 Hz (P less than 0.001) with both types of stimulation. In the sciatic nerve-tibialis anterior preparation the force of contraction was only higher at 3 and 5 Hz (P less than 0.01). The similar magnitude of the increase of the force of contraction during direct and indirect stimulation indicates that it is caused by facilitation of the contraction of the muscle fibers.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547328 TI - Fluoroquinolones for malaria: the newest kid on the block? PMID- 2547327 TI - Hemodynamic effects of experimental iron poisoning. AB - The hemodynamic effects of severe iron poisoning were studied in five mongrel dogs. Anesthetized animals were instrumented with arterial, venous, and pulmonary artery thermodilution catheters. Iron intoxication was induced by orogastric administration of ferrous sulfate (600 mg/kg elemental iron). Pulmonary artery wedge pressure values were maintained near preintoxication values by saline infusion, and sodium bicarbonate (1.5 mEq/kg/dose) was given for pH less than 7.25. Hourly hemodynamic measurements were obtained for five hours. Cardiac output, mean arterial pressure, pH, and heart rate decreased significantly (P less than .05), whereas systemic vascular resistance, left ventricular stroke work, and oxygen consumption did not change. All animals developed metabolic acidosis despite saline (3.6 +/- 0.9 L, mean +/- SD) and bicarbonate administration (4.2 +/- 0.8 mEq/kg). These findings suggest that decreased cardiac output was partially due to decreased heart rate but not to decreased preload or abnormal left ventricular afterload. Alkali therapy and maintenance of oxygen consumption did not prevent development of metabolic acidosis. PMID- 2547329 TI - Co-infection with human immunodeficiency virus-type 1 (HIV-1) and cytomegalovirus in two intravenous drug users. AB - A mononucleosis-like illness is frequently recognized retrospectively as the first manifestation of infection with human immunodeficiency virus-type 1 (HIV 1). This acute but transient retroviral syndrome may include symptoms such as malaise, fever, sweats, myalgia, arthralgia, maculopapular rash, diarrhea, and lymphocytic meningitis. We observed two intravenous drug users who developed a severe, febrile illness with subsequent oral thrush (one also had biopsy-proven esophageal candidiasis). Both patients had weight loss, arthralgia, myalgia, and fatigue. These symptoms occurred two weeks after needle-sharing and persisted for 7 weeks in one patient and 10 weeks in the other. Both patients had serologic evidence for both acute HIV-1 and cytomegalovirus infection. Cytomegalovirus enhances HIV-1 replication in vitro, presumably by stimulating HIV-1 gene expression. Thus, the observed syndrome suggests that this viral interaction may be clinically significant because it appears to cause severe additional morbidity, which is not typical for primary infection with HIV-1. After 6 months of follow-up, one patient is completely asymptomatic but shows markedly reduced CD4+ lymphocytes. The other patient developed persistent lymphadenopathy after the acute illness, but is feeling well 21 months after infection. PMID- 2547330 TI - [Treatment of chronic refractory cardiac insufficiency with enalapril]. AB - The objective is here to study the long and intermediate term clinical and haemodynamic effects of enalapril during chronic heart failure resistant to the classic digitalis-diuretics treatment. The study involves 16 patients (12 males and 4 females), with a mean age of 50 years. Before being given enalapril, 12 patients were at stage IV and 4 patients at stage II of the NYHA; the mean capillary pressure was quite elevated (30 +/- 6.3 mmHg), the cardiac index has collapsed (2.12 +/- 0.38 l.min.m2) and the stroke fraction (SF) is 0.28 +/- 0.08. At the 1st month control, there is a definite functional and haemodynamic improvement of the pre-charge as well as the post-charge. This improvement is still present at 6 months. The ventricular function is improved (SF = 0.38 +/- 0.13; p less than 0.001). The clinical tolerance of enalapril is excellent and the only adverse reaction is a transient deterioration of the renal function in a patient with diabetic glomerulopathy. PMID- 2547331 TI - Clinical use of gynaecologic tumour markers. AB - Tumour markers have proved to be important in certain types of gynaecological cancer. The treatment of chorionic cancer is largely based on changes in the serum levels of human choriogonadotropin (hCG). There are no other markers of equal utility, but some new markers for ovarian cancer show promise of becoming clinically important in the follow-up of patients. Assay of CA 125 has become a routine method in the follow-up of nonmucinous ovarian cancer, and tumour associated trypsin inhibitor (TATI) shows promise of being useful for mucinous ovarian cancer. In the rare ovarian embryonal tumours hCG and alphafetoprotein (AFP) are often useful. For other types of gynaecological cancer, there are no equally useful markers, but CA 125 is relatively useful in endometrial cancer and SCC (squamous cell carcinoma-associated antigen) in cervical cancer. Carcinoembryonic antigen (CEA) is occasionally useful in ovarian and cervical cancer. PMID- 2547332 TI - Wilms' tumour and related syndromes. A unifying theory. AB - The major role of somatic crossing-over in the aetiology of Wilms' tumour and other forms of cancer is emphasized. A model is proposed to show that tumour development is due to a cis-trans disruptive effect of somatic crossing-over on the maintenance of differential parental imprinting. PMID- 2547333 TI - Comparison of fecal microflora of elderly persons in rural and urban areas of Japan. AB - The Fecal microflora of 15 healthy elderly persons with a median age of 84 years in a rural area whose inhabitants tend to be long-lived (Yuzurihara-area, Uenohara, Yamanashi Prefecture) was compared with the microflora of individuals with a median age of 68 years in an urban area (Tokyo). The diet of the elderly persons in the Yuzurihara area is characterized by a high intake of dietary fiber. Total numbers of anaerobic bacteria were significantly smaller in the elderly persons in the Yuzurihara area than those in the Tokyo area. A significantly large number of bifidobacteria, but not of lecithinase-negative clostridia, was observed in the elderly persons in the Yuzurihara area. Large numbers and high incidences of bacilli and lecithinase-positive clostridia (mainly Clostridium perfringens) were found in the elderly persons in the Tokyo area. Twenty-five genera and over 81 species were isolated from the elderly persons in the Yuzurihara area, and 25 genera and over 92 species were isolated from the elderly persons in the Tokyo area. Furthermore, significantly larger numbers of Bifidobacterium adolescentis and Fusobacterium mortiferum strains were found in the Yuzurihara group, but significant reductions in the Bacteroides buccae-oris group, B. thetaiotaomicron, Bacteroides spp., C. coccoides, C. paraputrificum, and Clostridium spp. were observed in the same group. A significantly higher isolation rate of Bacillus subtilis was observed in the elderly persons in the Tokyo area. The difference in the fecal microflora between elderly persons in Yuzurihara and those in the Tokyo area might be due to a difference in the intake of dietary fiber. PMID- 2547334 TI - Solubilization and renaturation of overexpressed aggregates of mutant tryptophan synthase alpha-subunits. AB - Certain Escherichia coli tryptophan synthase mutant alpha-subunits encoded from mutagenized trpA-containing plasmids were overexpressed as insoluble aggregates which were seen as large, intracellular inclusion bodies. The insoluble aggregates were solubilized to various degrees by several neutral, chaotropic salts. The order of effectiveness of these salts (KSCN, NaI greater than NaNO3, LiBr greater than CaCl2) followed that for the Hofmeister series. Optimum conditions for the use of KSCN resulted in a maximum 70 to 75% solubilization of the aggregate forms for all mutant alpha-subunits examined. Removal of KSCN by dialysis resulted in the recovery of biological activity and of certain characteristic structural properties. Such salts may be a useful alternative for other recombinant protein aggregates which resist complete renaturation by commonly used treatments with guanidine or urea. PMID- 2547336 TI - Environmental significance of the potential for mer(Tn21)-mediated reduction of Hg2+ to Hg0 in natural waters. AB - The role of mer(Tn21) in the adaptation of aquatic microbial communities to Hg2+ was investigated. Elemental mercury was the sole product of Hg2+ volatilization by freshwater and saline water microbial communities. Bacterial activity was responsible for biotransformation because most microeucaryotes did not survive the exposure conditions, and removal of larger microbes (greater than 1 micromole) from adapted communities did not significantly (P greater than 0.01) reduce Hg2+ volatilization rates. DNA sequences homologous to mer(Tn21) were found in 50% of Hg2+-resistant bacterial strains representing two freshwater communities, but in only 12% of strains representing two saline communities (the difference was highly significant; P less than 0.001). Thus, mer(Tn21) played a significant role in Hg2+ resistance among strains isolated from fresh waters, in which microbial activity had a limited role in Hg2+ volatilization. In saline water environments in which microbially mediated volatilization was the major mechanism of Hg2+ loss, other bacterial genes coded for this biotransformation. PMID- 2547335 TI - Hydrogenase synthesis in Bradyrhizobium japonicum Hupc mutants is altered in sensitivity to DNA gyrase inhibitors. AB - In the Hupc mutants of Bradyrhizobium japonicum SR, regulation of expression of hydrogenase is altered; the mutants synthesize hydrogenase constitutively in the presence of atmospheric levels of oxygen. The DNA gyrase inhibitors nalidixic acid, novobiocin, and coumermycin were used to inhibit growth of wild-type and mutant cells. For each inhibitor tested, growth of mutant and wild-type strains was equally sensitive. However, in contrast to the wild type, the Hupc mutants synthesized hydrogenase in the presence of high levels of any inhibitor. Cells were incubated with the drugs and simultaneously labeled with 14C-labeled amino acids, and hydrogenase was immunoprecipitated with antibody to the large subunit of the enzyme. Fluorograms of antibody blots then were scanned to determine the relative amount of hydrogenase (large subunit) synthesized in the presence or absence of the gyrase inhibitors. The amount of hydrogenase synthesized by the Hupc mutants in the presence of 300 micrograms of nalidixic acid per ml was near the level of enzyme synthesized in the absence of the inhibitor. No hydrogenase was detected in antibody blots of wild-type cultures which were derepressed for hydrogenase in the presence of 100 micrograms of coumermycin or novobiocin per ml. In contrast, hydrogenase was synthesized by the Hupc mutants in the presence of 100 micrograms of either drug per ml. The amount synthesized ranged from 5 to 32% and 20 to 49%, respectively, of that in the absence of those inhibitors, but nevertheless, hydrogenase synthesis was detected in all of the mutants examined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547338 TI - The one-electron oxidation of porphyrins to porphyrin pi-cation radicals by peroxidases: an electron spin resonance investigation. AB - For the first time, the enzymatic one-electron oxidation of several naturally occurring and synthetic water-soluble porphyrins by peroxidases was investigated by ESR and optical spectroscopy. The ESR spectra of the free radical metabolites of the porphyrins were singlets (g = 2.0024, delta H = 2-3 G), which we assigned to their respective porphyrin pi-cation free radicals. Several porphyrins were investigated and ranked by the intensity of their ESR spectra (coproporphyrin III greater than coproporphyrin I greater than deuteroporphyrin IX greater than mesoporphyrin IX greater than Photofrin II greater than protoporphyrin IX greater than uroporphyrin I greater than uroporphyrin III greater than hematoporphyrin IX). The porphyrins were oxidized by several peroxidases (horseradish peroxidase, lactoperoxidase, and myeloperoxidase), yielding the same type of ESR spectra. From these results, we conclude that porphyrins are substrates for peroxidases. The changes in the visible absorbance spectra of the porphyrins during enzymatic oxidation were monitored. The two-electron oxidation product, which was assigned to the dihydroxyporphyrin, was detected as an intermediate of the oxidation process. The optical spectrum of the porphyrin pi-cation free radical was not detected, probably due to its low steady-state concentration. PMID- 2547337 TI - Factors relevant in bacterial pyrroloquinoline quinone production. AB - Quinoprotein content and levels of external pyrroloquinoline quinone (PQQ) were determined for several bacteria under a variety of growth conditions. From these data and those from the literature, a number of factors can be indicated which are relevant for PQQ production. Synthesis of PQQ is only started if synthesis of a quinoprotein occurs, but quinoprotein synthesis does not depend on PQQ synthesis. The presence of quinoprotein substrates is not necessary for quinoprotein and PQQ syntheses. Although the extent of PQQ production was determined by the type of organism and quinoprotein produced, coordination between quinoprotein and PQQ syntheses is loose, since underproduction and overproduction of PQQ with respect to quinoprotein were observed. The results can be interpreted to indicate that quinoprotein synthesis depends on the growth rate whereas PQQ synthesis does not. In that view, the highest PQQ production can be achieved under limiting growth conditions, as was shown indeed by the much higher levels of PQQ produced in fed-batch cultures compared with those produced in batch experiments. The presence of nucleophiles, especially amino acids, in culture media may cause losses of PQQ due to transformation into biologically inactive compounds. Some organisms continued to synthesize PQQ de novo when this cofactor was administered exogenously. Most probably PQQ cannot be taken up by either passive diffusion or active transport mechanisms and is therefore not able to exert feedback regulation on its biosynthesis in these organisms. PMID- 2547339 TI - Preparation and characterization of monoclonal antibodies against human myeloperoxidase. AB - We established 11 hybridomas producing monoclonal antibodies (MoAbs), designated AM, against human myeloperoxidase (MPO), by immunizing mice with the three forms of MPO (I, II, and III) purified from healthy human polymorphonuclear leukocytes (PMN) and characterized the specificity of the AM MoAbs. Ten of the AM MoAbs reacted similarly to each of the three forms using an enzyme-linked immunosorbent assay. When a cetyltrimethylammonium bromide (CETAB) extract of human PMN was electrophoresed in a CETAB polyacrylamide gel and transferred to a nitrocellulose filter, IgG1 class AM MoAbs immunostained only the MPO band of the proteins of the extract. In addition, the AM MoAbs reacted to two radioactive bands of 94 and 92 kDa in a HL-60 cell lysate labeled with [35S]methionine for 1 h. After a chase period of 24 h, these bands were replaced by four radioactive bands of 64.5, 43, 16.7, and 13.4 kDa, demonstrating that the MoAbs recognize not only mature MPO but also the MPO precursors of 94 and 92 kDa. The data also indicated that the two major bands of 64.5 and 13.4 kDa corresponded to heavy and light chains of mature MPO, respectively, and the additive intermediate bands of 43 and 16.7 kDa were MPO-related proteins. Moreover, AM MoAbs reacted to a similar extent to the deglycosylated form of MPO III with endo-beta-N-acetylglucosaminidase H (Endo-H). Thus, IgG1 class AM MoAbs recognized MPO with high specificity and reacted to the structure which is commonly conserved in the three mature forms of MPO (I, II, and III), MPO precursors, and deglycosylated MPO with Endo-H. AM MoAbs also specifically reacted to PMN and/or monocytes but did not react to lymphocytes when the cell staining method was used. PMID- 2547340 TI - Electron spin resonance measurements of the effect of ionophores on the transmembrane pH gradient of an acidophilic bacterium. AB - The delta pH in ionophore-treated cells of an acidophile has been determined by electron spin resonance spectroscopy. The values obtained were comparable to those obtained using the more conventional techniques involving radiolabeled probes. No binding of the spin-labeled probe was observed as determined by two independent control experiments and by the characteristics of the probe signal. These results led us to conclude that the delta pH measured in protonophore/ionophore-treated cells is a result of a Donnan potential, which may be a physical property of all intact bacterial cells at low pH values. PMID- 2547341 TI - Nutritional and hormonal regulation of mRNA abundance for arginine biosynthetic enzymes in kidney. AB - Argininosuccinate synthetase and argininosuccinate lyase catalyze the synthesis of arginine from citrulline in kidney and also serve as components of the urea cycle in liver of ureotelic animals. Dietary and hormonal regulation of mRNAs encoding these enzymes have been well studied in liver but not in kidney. Messenger RNAs for these enzymes are localized within the renal cortex. Starvation and extreme variations in dietary protein content (0% vs 60% casein) produced 2.6- to 3.5-fold increases in mRNA abundance for these two enzymes in rat kidney. Argininosuccinate lyase mRNA was not induced by dibutyryl cAMP, dexamethasone, or a combination of the two agents. In contrast, argininosuccinate synthetase mRNA was induced 2-fold by dibutyryl cAMP but was unresponsive to dexamethasone. Thus, diet and hormones regulate levels of these mRNAs in rat kidney, but the responses are both qualitatively and quantitatively distinct from the responses previously reported for rat liver. PMID- 2547342 TI - Stabilization of soluble active rat liver glucagon receptor. AB - Active glucagon receptor was solubilized with 3-(3 cholamidopropyl)dimethylammonio-1-propanesulfonate (Chaps) from rat liver plasma membranes but rapidly (less than 8 h) lost activity. Either inclusion of 1X Hanks' balanced salt solution in the 3 mM Chaps solubilization buffer or its addition after solubilization increased the percentage of total binding attributable to specific glucagon binding from approximately 10 to greater than 80%; of great importance, it increased the stability from near zero binding at 8 h to 50% binding at 48 h (4 degrees C). Of the Hanks' solution components, either NaCl (137 mM) or CaCl2 (1.26 mM) was effective in increasing specific binding to approximately 70 and 60% respectively: Mg salts were ineffective. Soluble receptor binding activity was assayed by dextran-coated charcoal adsorption of free hormone. The assay is rapid, simple, and reproducible. It is suitable for monitoring receptor activity during purification and molecular characterization. Competition binding studies gave an IC50 value of 10-20 nM (slope factor approximately 1), with or without GTP. Dissociation assays revealed GTP sensitivity when receptors were solubilized either as glucagon-receptor complexes or free receptor. Active glucagon-receptor complexes could be eluted from wheat germ lectin-agarose: neither concanavalin A-agarose nor soybean agglutinin agarose bind receptor. A glucagon degrading activity which co-solubilized with the receptor but did not require detergent for extraction was distinguishable from the soluble receptor not only by solubility but also by its heat stability (30 degrees C), its inhibition by bacitracin, its affinity for glucagon, its retention of activity for at least 1 week at 4 degrees C, and its size. PMID- 2547343 TI - Comparison of receptor binding, biological activity, and in vivo tracer kinetics for 1,25-dihydroxyvitamin D3, 1,25-dihydroxyvitamin D2, and its 24 epimer. AB - Scatchard analyses of 1,25-dihydroxyvitamin D receptors (VDR) from chick and rat intestine, bovine thymus, pig kidney cells (LLC-PK1), and human breast cancer cells (T-47D) demonstrated that 1,25-dihydroxyvitamin D3 (1,25-D3) and 1,25 dihydroxyvitamin D2 (1,25-D2) had equal affinities for VDR. 24-Epi-1,25 dihydroxyvitamin D2 (24-epi-1,25-D2) exhibited affinities for VDR equal to that of 1,25-D2 and 1,25-D3 in most of these tissues. Scatchard analysis with 24-epi [3H]1,25-D2 underestimated total VDR by 50-70% in rat intestine, LLC-PK1, and T 47D cells. The biological activity of 24-epi-1,25-D2 was found to be only 30-70% of 1,25-D3 and 1,25-D2 as determined by in vivo induction of intestinal calcium transport and bone calcium resorption in the rat and in vitro induction of 23- and 24-hydroxylase activities in T-47D cells. In vivo tracer kinetic studies demonstrated that in the rat 1,25-D3 and 1,25-D2 kinetics were similar, whereas 24-epi-1,25-D2 had a 25% shorter plasma half-life and was cleared from the body 2.8 times faster than the natural hormones. This more rapid clearance of 24-epi 1,25-D2 along with reduced VDR binding appears to explain the reduced biological activity of 24-epi-1,25-D2. Our data clearly demonstrate that although there are differences in side chain structure between 1,25-D2 and 1,25-D3, the VDR binding, biological activity, and whole body tracer kinetics of these two metabolites are virtually identical. However, movement of the 28 methyl of 1,25-D2 from its natural S configuration to the R configuration significantly alters the activity of this hormone. PMID- 2547344 TI - Eruptive syringoma. AB - Eruptive syringoma is a rare variant of syringoma that appears on anterior surfaces of the body, including the neck, chest, and arms. Textbooks state that this eccrine-derived tumor arises at puberty. We describe four cases of eruptive syringoma that began in childhood. We review the literature on this entity and suggest that the disorder typically presents between the ages of 4 and 10 years. Eruptive syringoma is a benign tumor that should be considered among the papular dermatoses of childhood. PMID- 2547345 TI - Recurrent facial nodule. Chondroid syringoma. PMID- 2547347 TI - The malignancy of large cell lung carcinomas. PMID- 2547346 TI - Immunohistochemical assessment of estrogen and progesterone receptors in stored imprints and cryostat sections of breast carcinomas. AB - A peroxidase-antiperoxidase technique was used to visualize estrogen and progesterone receptors in stored imprints and cryostat sections of breast carcinomas that were prepared at the time of biopsy or frozen section diagnosis. This was done to provide an alternate technique for the assessment of the receptor status of tumors that could not be adequately assayed with other biochemical or immunocytological methods. Fixation in Zamboni's fixative followed by passage through cold methanol and acetone before storage at -80 C insured good preservation of the receptor proteins over extended periods of time (up to 56 weeks). Immunostaining of these stored preparations with monoclonal antibodies against estrogen receptor (H222) and progesterone receptor (JZB39 and KD68) showed a high degree of correspondence with immunocytochemical assays (ER-ICA and PR-ICA) and biochemical analysis. This technique is easy to perform and provides reliable information, even in tumors that are too small and/or ill defined to permit separate sampling for receptor assays. PMID- 2547348 TI - Large cell carcinoma of the lung: a highly aggressive tumor with dismal prognosis. AB - A retrospective review was made of 96 consecutive patients with large cell carcinoma of the lung admitted to Emory University Hospital over 10 years. Only 10 patients were seen with stage I lesions favorable for resection. The remainder were treated primarily with irradiation or chemotherapy. Mean survival for clinical stage I patients was 15.9 months; stage IIIA patients, 7.9 months; stage IIIB patients, 7.1 months; and stage IV patients, 5.8 months. Only 1 patient survived for 5 years. This distinct and highly aggressive form of lung cancer most commonly is seen at an advanced stage and is associated with an unusually dismal prognosis regardless of the method of treatment employed. PMID- 2547350 TI - Angiomyxoma of the umbilical cord with massive cystic degeneration of Wharton's jelly. AB - Tumors of the umbilical cord, and especially angiomyxomas, are extremely unusual placental lesions. Sonography three weeks after the finding of a slightly elevated amniotic fluid alpha-fetoprotein, detected a cystic lesion of the umbilical cord in a 35-year-old woman at 19.5 weeks gestation. Two prior sonograms had been normal (16.2 and 17.5 weeks gestational age). After delivery of a normal female infant at 38.5 weeks, a pathologic examination revealed an angiomyxoma of the umbilical cord with massive cystic degeneration of Wharton's jelly. The tumor was twice the size of cases cited previously and cystic degeneration was massive. Serial sonography afforded a unique opportunity to study the development of this lesion in utero. PMID- 2547349 TI - Cardiovascular action of penta-O-ethylquercetin in the rat. AB - Intravenous (i.v.) administration of penta-O-ethylquercetin (PQ) dissolved in glycerolformal (GF) induced a rapid and transient fall in arterial blood pressure of rats. The mechanism appears rather complex. Beside its inhibitory properties toward cardiac cyclic AMP phosphodiesterase, the drug seems to lower the adrenergic tone, thus acting both at the central (stimulation of the cardiomoderator medullary center) and peripheral (partial blockade of postsynaptic alpha-adrenergic receptors, decrease in noradrenaline liberation) levels. Repeated i.v. administration of variable dosages of PQ caused a decrease in the hypotensive effect of PQ/GF which might be related to a tachyphylaxy phenomenon. In marked contrast, continuous i.v. infusion of PQ/GF progressively increased arterial blood pressure, which very likely involves a quite different mechanism of action. PMID- 2547351 TI - Congenital idiopathic corneal endotheliopathy. AB - Two unrelated boys had a history of bilateral corneal clouding at birth following uncomplicated full-term gestations and spontaneous vaginal deliveries (without forceps). Clinical examinations disclosed bilateral corneal edema, no inflammation, and normal intraocular pressures. There was no history of similarly affected family members. The patients underwent penetrating keratoplasty at ages 4 months (patient 1) and 12 years (patient 2). Light and electron microscopic studies of the corneal buttons from both patients revealed areas of degeneration of the endothelium and separation of rounded endothelial cells. The morphologic features were strikingly similar to those in two acquired forms of corneal disorders--autoimmune endotheliopathy and "acute endotheliitis." Immunocytologic and in situ hybridization studies for herpes simplex virus were not consistent with either productive or latent corneal infection. Ultrastructural changes in Descemet's membrane reflect delayed or abnormal development of the postnatal nonbanded layer in patients 1 and 2, respectively. These suggest an intrauterine insult that resulted in endothelial dysfunction. The histologic and ultrastructural features of these two congenital cases are not typical of those seen in any of the recognized causes of congenital corneal clouding. We propose that these cases represent a unique congenital corneal endotheliopathy of undetermined origin. PMID- 2547352 TI - Oral acyclovir reduces the incidence of recurrent herpes simplex keratitis in rabbits after penetrating keratoplasty. AB - To determine if acyclovir sodium prevents postoperative herpes simplex virus type 1 (HSV-1) recurrences, 21 rabbits harboring latent HSV-1 underwent uniocular autograft penetrating keratoplasty. All operated-on eyes were treated with topical and subconjunctival dexamethasone sodium phosphate. Ten of the 21 rabbits also received oral acyclovir (intravenous acyclovir was given at the time of surgery). Postoperatively, 9 (82%) of 11 operated-on eyes in rabbits not treated with acyclovir had positive HSV-1 ocular cultures. In acyclovir-treated rabbits, however, none of the 10 operated-on eyes had positive ocular cultures. In addition, 9 (82%) of 11 of the operated-on eyes had geographic ulcers develop in the non-acyclovir-treated rabbits, compared with 1 (10%) of 10 in the acyclovir treated rabbits. Finally, stromal keratitis appeared in 5 (56%) of 9 of the operated-on eyes in non-acyclovir-treated rabbits and 1 (12%) of 8 of the operated-on eyes in acyclovir-treated rabbits. The results of this study indicate that acyclovir significantly lowered the incidence of HSV-1 ocular shedding, geographic ulceration, and stromal keratitis in a rabbit autograft penetrating keratoplasty model. PMID- 2547353 TI - Epstein-Barr virus infection in cultured non-malignant epithelial cells from human nasopharyngeal mucosa. AB - Epstein-Barr virus infected cultured epithelial cells from the human nasopharyngeal mucosa only, when the cells were maintained under conditions preventing terminal differentiation. The infection of these cells resulted in a marked cytopathology resembling that morphology induced in cell cultures by other members of the herpesvirus group. PMID- 2547354 TI - Adenoid cystic carcinoma of the cervix: a report and review of 3 cases. AB - Adenoid cystic carcinoma was reported in 3 Japanese women. In all cases, the condition followed the same rapid pattern of deterioration described in previous literature. The characteristic cribriform pattern was observed in all patients under a microscope, even in small parts of the tumor. This finding of this tumor may indicate that the adenoid cystic carcinoma is not identical to, but a variation of an undifferentiated carcinoma derived from the reserve cell. PMID- 2547355 TI - A flavor paired with lithium chloride blocks the formation of a pentobarbital lithium chloride association. AB - Thirsty rats were used in order to determine whether a vinegar solution, which had been paired with an injection of lithium chloride, could block the formation of an association between a pentobarbital- and a lithium chloride-induced state. During phase 1 the rats in the blocking group had a 2.0% vinegar solution paired with an injection of 240 mg/kg of lithium chloride, during phase 2 these rats were reexposed to the vinegar prior to each injection of 20 mg/kg of pentobarbital and 240 mg/kg of lithium chloride, and during phase 3 these rats were given access to a novel 0.75% saccharin solution and were injected with pentobarbital after saccharin removal. Animals with this history did not form an association between the pentobarbital- and lithium chloride-induced states during phase 2 as evidenced by their refusal to consume the saccharin solution over repeated pairings of saccharin with pentobarbital during phase 3. Control groups that received forward pairings of pentobarbital and lithium chloride, in the absence of a previously conditioned vinegar solution during phase 2, formed an association between pentobarbital and lithium chloride. These findings indicate that drug states and flavors can interfere with each others' capacity to predict the occurrence of lithium chloride. PMID- 2547356 TI - Cytochemical localization of alkaline phosphatase and Na+, K+-ATPase activities in the blood-brain barrier of Rana esculenta. AB - The ultrastructural distribution of alkaline phosphatase and Na+, K+-ATPase on the brain capillaries in Rana esculenta was investigated. Alkaline phosphatase activity appears both on the luminal and abluminal walls of the endothelial capillary cells; Na+, K+-ATPase is, instead, only present on the abluminal side. This different enzymatic distribution indicates that endothelial cells of the brain capillaries are polarized and the luminal and abluminal endothelial membranes are functionally different. The role of these two enzymatic activities is discussed in relation to the blood-brain barrier. PMID- 2547357 TI - Brain cytochrome oxidase activity of synaptic and nonsynaptic mitochondria during aging. AB - The activity of cytochrome c oxidase was studied in aging brain on non-synaptic and intra-synaptic mitochondria from frontal cerebral cortex, hippocampus and striatum of 4, 8, 12, 16, 20 and 24 month-old Sprague-Dawley rats. Specific activities of cytochrome oxidase were significantly higher in light synaptic mitochondria than in non-synaptic or heavy ones at all the ages examined. However, enzyme activity in light mitochondria from cerebral cortex remains unchanged during aging, being increased in hippocampus and striatum. These results indicate that aging affected not only the various cerebral area (macroheterogeneity), but also the different mitochondrial populations (subcellular heterogeneity). PMID- 2547358 TI - Information transfer in the regulation of striated muscle contraction. AB - A brief review of current views on the regulation of striated muscle contraction is presented with emphasis on the transmission of the effect of calcium binding to the N-terminal domain of troponin C to the other components of the regulatory system. Results of recent work on chemical modification of troponin C as well as studies on crosslinking among thin filament components are discussed. PMID- 2547360 TI - Ca-channel currents in isolated frog ventricular cells are increased by thrombin. AB - Effects of the proteolytic enzyme thrombin in the modulation of cardiac Ca channel currents were examined in single ventricular cells from frog myocardium, using the whole-cell voltage clamp technique (1). Application of 3.8 . 10(-9) M thrombin to the bath increased the peak of the Ca-channel current by 84 +/- 35% (8 cells). Hirudin (31.10(-9)M), a specific thrombin inhibitor, blocked the thrombin-induced increase of this current. The increase in the current can be made responsible for the measured positive inotropic effects on frog heart of thrombin. PMID- 2547359 TI - Calcium channels in skeletal muscle fibres of the frog. AB - The vaseline gap voltage clamp technique was used to study slow and fast calcium currents in phasic and tonic muscle fibres of the frogs Rana temporaria (R.t.) and Xenopus laevis (X.l.). At physiological Ca2+ and Na+ concentrations three inward currents could be recorded in R.t. twitch muscle fibres: a fast Ca current, a slow Ca current, and a non-specific (INS) current carried by Na ions. The last current appeared as a sole component at a low threshold membrane potential (MP -55 mV) and was present until the slow Ca current appeared. In tonic R.t. muscle and twitch X. laevis fibres only the fast and the slow Ca currents could be recorded; however the inactivation of the slow Ca current was very prolonged in the presence of Na ions. A modulatory effect of Na ions on Ca2+ currents was observed. In phasic fibres the fast Ca currents inactivated either very slowly or rapidly. PMID- 2547361 TI - Subconductance states in cardiac sodium channels. AB - Single Na channel currents were studied in cell-attached patches from guinea-pig ventricular myocytes in the presence of DPI 201-106. In addition to openings with a single channel conductance of 15 pS, two other types of events occur that are characterized by a conductance of 5 and 8 pS. The low conductance state is less sensitive to block by TTX than the higher conductance states. The medium conductance state has a reversal potential which is similar to the high conductance channel. It is argued that the low conductance channel is a second type of Na channel, whereas the medium conductance channel appears to be a low conductance substate of the normal Na channels. PMID- 2547362 TI - Ca2+-transport by smooth muscle membranes and its regulation. AB - In smooth muscle cells two distinct Ca2+-pumps with a different subcellular localization can be demonstrated. A plasma-membrane localized Ca2+-pump with a relative molecular weight (Mr) of 140 kDa resembles the Ca2+-pump of the erythrocyte plasma membrane in the sensitivity of its phospho-intermediate towards La3+, in its calmodulin-binding capacity and in its antigenic properties. A second Ca2+-pump with a Mr of 100 kDa is situated in the endoplasmic reticulum. On the basis of its antigenicity and the degradation pattern of its phospho intermediate the endoplasmic-reticulum Ca2+-pump is found to be homologous to the sarcoplasmic-reticulum Ca2+-pump of cardiac muscle and slow twitch skeletal muscle, but it clearly differs from the Ca2+-pump present in the sarcoplasmic reticulum of fast skeletal muscle. The endoplasmic-reticulum and the plasma membrane Ca2+-pumps are present in both visceral and vascular smooth muscle, but tissue-and species-dependent differences in their relative amount have been observed. The endoplasmic-reticulum Ca2+-pump is regulated via phospholamban. Phosphorylation of this regulatory protein by cAMP-dependent as well as by cGMP dependent protein kinase stimulates the endoplasmic-reticulum Ca2+-pump. On the other hand, the activity of the plasmalemmal Ca2+-pump is modulated by calmodulin, negatively charged phospholipids and membrane-receptor-binding agonists. cGMP-dependent protein kinase also exerts a stimulatory effect on the plasmalemmal Ca2+-pump. However, cGMP-dependent protein kinase does not directly phosphorylate the plasmalemmal Ca2+-pump, but by activating a phosphatidyl inositol kinase it promotes the formation of phosphatidyl-inositol monophosphate which then acts as the final stimulator of the Ca2+-pump. PMID- 2547363 TI - Comparison of Ca2+-extrusion systems in the myometrial plasma membrane of pregnant and non-pregnant rats. AB - Plasma membrane derived from the myometrial tissues of pregnant and non-pregnant rats accumulates Ca2+ both by Na+ and by ATP-dependent ways. Sodium-dependent Ca2+-uptake in plasma membranes obtained from 20 days pregnant rats reveals higher Ca2+ sensitivity than in plasma membrane of non-pregnant rats. Ca2+ concentration required for half maximal Ca2+-transport was 5.7 +/- 0.5 microM and 11.8 +/- 0.6 microM in plasma membrane of pregnant and non-pregnant rats, respectively. The initial rate of sodium-dependent Ca2+-uptake also differed in membrane preparations from pregnant and non-pregnant animals. ATP-dependent Ca2+ uptake in myometrial plasma membrane of pregnant rats was higher than in non pregnant ones. The difference was abolished by calmodulin antagonist drugs. In calmodulin depleted preparations the administration of exogenous calmodulin restored Ca2+-uptake to the rate measured in calmodulin not-depleted ones. Membrane phosphorylation by endogenous protein kinases proved to be also different in the two preparations. PMID- 2547364 TI - Extracellular appearance of calpain and calpastatin in the synovial fluid of the knee joint. AB - Extracellular location of calpain and calpastatin was demonstrated in the cell free synovial fluid obtained from the knee joint of healthy adult humans and several patients with rheumatoid arthritis. Calpains I and II and a few molecular species of calpastatin were identified by chromatographies on DEAE-cellulose and on Ultrogel AcA 34 columns as well as by immunoelectrophoretic blot analysis. Calpains I and II in the synovial fluid of the patients increased 6.7 times and 3.5 times, respectively, compared with those of the control subjects. With the patients, shortening of the heavy subunits of calpains was noted. Calpastatin also increased in the patients, but it showed rather extensive fragmentation. PMID- 2547365 TI - Fetal and adult sheep adrenal cortical cells contain glucocorticoid receptors. AB - Specific receptors for glucocorticoids were identified in the fetal and adult sheep adrenal cortex by a whole-cell binding assay using [3H]triamcinolone acetonide ([3H]TA) as the radiolabelled ligand. [3H]TA binding sites were saturable and of high affinity, with dissociation constant (Kd) of 2-3nM. Scatchard analysis revealed a single class of binding sites with a binding capacity (Bmax) of 207 and 5 fmol/10(6) cells for d100 fetuses and adults, respectively. By single point analyses at saturating [3H]TA concentration, we found that glucocorticoid receptors (GR) were present in the fetal adrenal cortex as early as d60. Highest concentrations were found at d100-110. GR decreased to d125, then increased to term (approx. d145) before declining again in newborn and adult animals. This demonstration of glucocorticoid receptors in ovine fetal adrenal cortical cells provides a mechanistic basis for the concept that glucocorticoids may act, perhaps in a paracrine or autocrine fashion, to influence adrenocorticotropin (ACTH)-induced activation of fetal adrenal function and the events leading to parturition. PMID- 2547366 TI - Transformation of leukotriene A4 to lipoxins by rat kidney mesangial cell. AB - Incubation of rat mesangial cells with leukotriene A4 in the presence of calcium ionophore A23187 led to a substrate dependent formation of lipoxin and its isomers. The major metabolite coeluted with authentic lipoxin A4 (LXA4) and lipoxin B4 (LXB4) in RP-HPLC system, and possessed a characteristic U.V. spectrum and C-value which were identical to authentic standards. GC/MS analysis on LXA4 further demonstrates that the mesangial cell derived LXA4 was identical to that reported by Serhan et al. (1) as LXA4 [5(S), 6,(R), 15(S)-trihydroxy7,9,13-trans 11-cis-eicosatetraenoic acid]. The formation of LXA4 was linear with substrate (LTA4) concentration. No similar products occurred in boiled controls. Incubation of mesangial cell with 15-HPETE failed to produce any lipoxin-like material. The absence of LX-like substance following incubation of 15-HPETE with mesangial cells suggested that 5-lipoxygenase activity is not expressed in mesangial cells under these conditions. The generation of LXA4 from LTA4 in mesangial cells suggested that there is an active 15- or 12- lipoxygenase activity in the kidney. The production of LX may play an important role in the regulation of renal function and the response to inflammatory stimuli. PMID- 2547367 TI - Transcription of 5S RNA genes containing insertion mutation and the assembly of preinitiation complexes. AB - The transcription of Xenopus 5S RNA genes containing insertions in and around the intragenic control region has been analyzed in a HeLa cell-free system. A 15 bp insertion within the intragenic region greatly diminished binding of TFIIIA and concomitantly reduced binding of TFIIIB and transcription in vitro. This study helps to define more precisely the promoter organization of Xenopus 5S RNA genes for the efficient interactions with transcription factors. PMID- 2547368 TI - Accumulation of cAMP in the cells of Candida tropicalis at an early stage of ethanol-induced filamentous growth and its prevention by myo-inositol. AB - Phosphatidylinositol metabolism is enhanced in the cells of Candida tropicalis Pk 233 at an early stage of filamentous growth caused by ethanol, and myo-inositol prevents the ethanol-induced changes in the metabolism and morphology [Uejima et al. (1987) FEBS Lett. 214, 127-129]. The accumulation of cAMP and an increase in adenylate cyclase activity were observed in the cells grown with ethanol to the mid-log phase. Myo-inositol abolished these effects of ethanol also. The activity of cAMP phosphodiesterase was affected by neither ethanol nor myo-inositol. These results suggest that the inositol phospholipid-linked and cAMP-linked signaling pathways may be involved in the mechanism of ethanol-induced filamentous growth of this yeast and also that myo-inositol would affect morphogenesis by controlling these pathways. PMID- 2547369 TI - Vector electron paramagnetic resonance spectroscopy with first and second harmonic displays of ferrihemoglobin. AB - Superimposed plots of electron paramagnetic resonance spectra with the first and second harmonic displays of ferrihemoglobin at pH 9.1 and 90 K were measured at 20 degree intervals of phase angle using a phase-sensitive detector. The high spin signal in the g = 6 region was observed in both displays, and a small splitting of the signal was found in the calculated amplitude spectrum of the second harmonic display, with g values of 5.95 and 6.05. Low spin signals were observed at g = 2.55, 2.25 and 1.82 in both harmonic displays. A signal in the g = 2.05 region was observed only in the second harmonic display. The signal is probably associated with the low spin spectrum; however, its origin is obscure. PMID- 2547370 TI - Effects of dietary eicosapentaenoate (20:5 n-3) on cardiac beta-adrenergic receptor activity in the marmoset monkey. AB - Eicosapentaenoic acid (EPA; 20:5 n-3; ethyl ester) in combination with atherogenic or non-atherogenic high fat diets was fed to young adult male marmoset monkeys for a period of 30 weeks. EPA markedly reduced the raised plasma cholesterol level evident when feeding an atherogenic diet but did not influence the cardiac membrane cholesterol-to-phospholipid ratio. EPA and its elongation product 22:5 n-3 was incorporated into cardiac membrane phospholipids at the expense of linoleic and arachidonic acids. Dietary EPA increased cardiac beta-AR affinity and reversed the decreased beta-AR affinity evident when feeding an atherogenic diet. PMID- 2547371 TI - Decreased binding of vasoactive intestinal peptide to intestinal epithelial cells from hypothyroid rats. AB - The binding of vasoactive intestinal peptide (VIP) and stimulation of adenylate cyclase by VIP were studied in intestinal epithelial cells during hypothyroidism. Experimental hypothyroidism was induced in rats by the administration of KC10(4). The binding capacity, but not the affinity, of VIP receptors decreased in the hypothyroid rats. Besides, the stimulation of cyclic AMP production by VIP was also diminished in cells from hypothyroid rats. These observations indicate a decrease of the responsiveness of intestinal epithelial cells to VIP in the hypothyroid status, suggesting a role of the peptide in the pathophysiologic mechanism of intestinal manifestations during hypothyroidism. PMID- 2547372 TI - Antibodies to an NH2-terminal myristoyl glycine moiety can detect NH2-terminal myristoylated proteins in the retrovirus-infected cells. AB - Novel antibodies were raised against a synthetic NH2-terminal myristoyl glycine moiety which is characteristic of N-myristoyl-proteins. Antisera raised against N myristoyl-Gly-hemocyanin reacted with N-myristoyl-Gly-[125I]albumin. The immunoreaction was competed for by albumin conjugated with N-myristoyl-glycine, while underivatized albumin had no effect. Of the [3H]myristate-labeled proteins detected, pp60v-src, which is a transforming protein of Rous sarcoma virus, and p19gag and p17gag, which are core proteins in the human T-cell leukemia virus and the human immunodeficiency virus, were identified as N-myristoylated proteins by the radioimmunoprecipitation analyses with the antibody. PMID- 2547373 TI - Sequence of the 5'-flanking region of the rat 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase gene: regulation by glucocorticoids. AB - Dexamethasone addition to cultured hepatocytes caused a 90-fold increase in mRNA for 6-phosphofructo 2-kinase/fructose-2,6-bisphosphatase. Glucocorticoid administration in vivo also increased the enzyme's mRNA in skeletal muscle by 3-4 fold. The sequence of the 5'-flanking region of the enzyme's gene revealed at least one consensus glucocorticoid response element. The amino acid sequence derived from a partial cDNA clone for the rat skeletal muscle bifunctional enzyme was identical to that of the liver isozyme except for an undetermined amount of N terminal sequence. It is concluded that the rat muscle and liver isozymes, which are postulated to be identical except for the N-terminal region, are both regulated by glucocorticoids. PMID- 2547374 TI - Mineralization induced by beta-glycerophosphate in cultures leads to a marked increase in collagenase synthesis by mouse osteogenic MC3T3-E1 cells under subsequent stimulation with heparin. AB - The clonally derived mouse osteoblast-like cell line MC3T3-E1 was shown to produce latent collagenase (approximately 0.2 units/ml) under stimulation with either heparin or parathyroid hormone in confluent cultures. However, it was found that MC3T3 E1 cultures which were first induced to undergo mineralization by the addition of beta-glycerophosphate and were subsequently stimulated with heparin showed an approximately ten-fold increase in collagenase synthesis. MC3T3 E1 cell collagenase from a small sample of serum-free culture medium was purified 49-fold to a specific activity of 200 units/mg protein with a yield of 14% by heparin-sepharose affinity chromatography and ion-exchange high performance liquid chromatography. This new mineralization-primed cell culture system may be a valuable model for the study of osteoblast collagenase. PMID- 2547375 TI - Induction of manganese superoxide dismutase by tumor necrosis factor in human breast cancer MCF-7 cell line and its TNF-resistant variant. AB - Tumor necrosis factor (TNF)-resistant variant of human mammary cancer MCF-7 cell line was isolated by stepwise selection. The final TNF-resistant variant Tnf-1000 showed more than 100-fold higher resistance than the parental MCF-7 cell. Saturation kinetics for 125I-TNF binding showed that TNF-1000 cells had similar TNF receptor numbers as MCF-7 cells, but of a lower affinity. Induction of superoxide dismutase (SOD) was compared between MCF-7 and Tnf-1000 cells treated with TNF: SOD scavenges potentially toxic superoxide radicals. TNF induced more mitochondrial manganese SOD (SODm) in MCF-7 than in Tnf-1000 whereas there appeared to be no significant induction of cytosolic copper/zinc SOD (SODc) by TNF in both MCF-7 and Tnf-1000 cell lines. Acquirement of TNF-resistance in MCF-7 cells might be correlated with expression of SODm. PMID- 2547376 TI - Insulin and lipolytic hormones stimulate the same phosphodiesterase isoform in rat adipose tissue. AB - Insulin sensitive phosphodiesterase from rat adipocytes is found in particulate fractions. Solubilisation of the enzyme with triton X-100 yields a preparation containing more than one phosphodiesterase activity as judged by its rate of thermal denaturation at 45 degrees C and by its non-linear kinetic plots. Immunoprecipitation of solubilised activity with a polyclonal antiserum raised against purified insulin-sensitive rat liver phosphodiesterase selected a form of the enzyme which showed a single exponential decay of enzyme activity when heated at 45 degrees C and linear low Km kinetics. Treatment of adipocytes with insulin ACTH, glucagon or isoproterenol stimulated the low Km particulate phosphodiesterase. The hormonal activation was retained following solubilisation and was also seen when activity was immunoprecipitated. It is suggested that all four hormones activate the same form of phosphodiesterase. PMID- 2547377 TI - Computer analysis of 1,25-dihydroxyvitamin D3-receptor regulated promoters: identification of a candidate D3-response element. AB - The receptor for the steroid hormone 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] belongs to a superfamily of steroid and triidothyronine (T3) receptors. As yet no 1,25(OH)2D3 response element (DRE) has been identified. Since the T3 and 1,25(OH)2D3 receptors are structurally homologous, we have used the nucleotide sequence of the T3 response element to carry out a computer search of the promoter region of several 1,25(OH)2D3 regulated genes. We report here one candidate DRE from the chick calbindin-D28K gene (AGCCCAATGGCTGAACA) and two candidate DRE's from the rat osteocalcin gene (TCCCCACTGGATGAGCG and CCTGCACTGGGTGAATGA). PMID- 2547378 TI - Bluetongue virus-17 fusion protein ns1 expressed in Escherichia coli by pUC vectors. AB - The cDNA coding for the major nonstructural protein, NS1, of bluetongue serotype 17 (BTV-17) was cloned previously. Using pUC plasmids, we have successfully expressed the NS1 protein in Escherichia coli as a LacZ-NS1 fusion protein. The recombinant NS1 protein reacted with rabbit anti-BTV-17 antiserum, and was thus immunologically indistinguishable from the native BTV-17 NS1 protein. This was the first bluetongue viral protein to be produced in a bacterial system. PMID- 2547379 TI - Indolpyruvic acid administration increases the brain content of kynurenic acid. Is this a new avenue to modulate excitatory amino acid receptors in vivo? AB - The possibility of changing the tissue content of kynurenic acid (KYNA), a tryptophan metabolite which acts as an antagonist of the excitatory amino acid receptors, was investigated by measuring its concentration in the brain, blood, liver and kidney of rats using a specific method based on ion exchange chromatography and HPLC. The administration of tryptophan (TRP) or of its keto analogue, indolpyruvic acid (IPA) (50-500 mg/kg i.p.), significantly increased, in a dose-dependent manner, the content of KYNA in various organs, including the brain. The increased brain content of KYNA after IPA administration could not be completely explained by considering that IPA may be transaminated to TRP and that the enzymes leading from TRP to KYNA are known. An alternative pathway of KYNA synthesis from IPA was therefore proposed. These findings indicate that it is possible to change the brain content of an endogenous glutamate antagonist. This could be a new avenue to modulate in vivo excitatory amino acid receptors. PMID- 2547380 TI - Monocrotaline-induced cardiopulmonary injury in rats. Modification by the neutrophil elastase inhibitor SC39026. AB - Rats were killed after 6 weeks of continuous ingestion of the pneumotoxic alkaloid monocrotaline (2.2 mg/kg/day), the neutrophil elastase inhibitor SC39026 (60 mg/kg/day), or both. Pulmonary reactions were evaluated by light and electron microscopy. Lung endothelial function was monitored by angiotensin converting enzyme (ACE) activity, plasminogen activator (PLA) activity, and prostacyclin (PGI2) and thromboxane (TXA2) production. Lung hydroxyproline content was measured as an index of interstitial fibrosis. Cardiac right ventricular hypertrophy was determined by the right ventricle to the left ventricle plus septum weight ratio (RV/LV + S). Rats receiving SC39026 alone did not differ significantly from untreated control animals with respect to any of the quantitative endpoints, although rarefaction of Type I pneumocytes was observed in the electron micrographs of these animals. Monocrotaline-treated rats, in contrast, developed a significant increase in RV/LV + S, and exhibited pulmonary edema, inflammation, fibrosis, and muscularization and occlusive mural thickening of the pulmonary small arteries and arterioles. These monocrotaline-induced structural changes were accompanied by decreased lung ACE and PLA activities, and increased PGI2 and TXA2 production, and by an increase in lung hydroxyproline content. Cotreatment with SC39026 ameliorated the monocrotaline-induced pulmonary vascular wall thickening and the cardiac right ventricular hypertrophy. These data suggest that inappropriate neutrophil elastase activity contributes to monocrotaline pulmonary vasculopathy and hypertension. On the other hand, cotreatment with SC39026 had no significant effect on the severity of the monocrotaline-induced lung inflammatory reaction, the pulmonary endothelial dysfunction, or the increase in lung hydroxyproline content. PMID- 2547382 TI - Presence of sigma and phencyclidine (PCP)-like receptors in membrane preparations of bovine adrenal medulla. AB - Receptor binding studies with [3H]-(+)SKF-10047 were carried out to characterize the putative sigma (sigma) and phencyclidine (PCP) receptors in membrane preparations of bovine adrenal medulla. Specific binding of the radiolabelled compound was observed after incubation with the membrane preparation at 37 degrees, the equilibrium being reached at 20 min and the maximal binding being observed with 0.6 mg/ml protein. Saturation binding studies were performed at equilibrium (30 min at 37 degrees with 0.5 mg/ml of membrane protein) in the presence of haloperidol (1 microM) or 1-[1-(2-thienyl) cyclohexyl] piperidine (TCP; 0.2 microM) to block sigma or PCP receptors, respectively. The binding of [3H]-(+)SKF-10047 was characterized by two distinct components. A high affinity binding site (haloperidol sensitive) had an apparent KD of 8.3 nM and a Bmax of 67 pmol/g protein. A lower affinity binding site (TCP sensitive) had an apparent KD of 32.7 nM and a Bmax of 83 pmol/g protein. The drug specificity of the high affinity binding site resembled that of the putative sigma receptor, being potently inhibited by haloperidol and pentazocine. The binding pharmacology of the low affinity site resembled that of the phencyclidine receptor, being potently displaced by TCP and PCP. The binding of [3H]-(+)SKF-10047 to both receptors showed marked stereoselectivity for the dextrorotatory (+) isomer of SKF-10047 and was insensitive to the receptor specific opioid ligands DAGO (mu), DSLET (delta) and U-69593 (kappa). These data indicate that bovine adrenal medulla contains sigma and PCP-like receptors. PMID- 2547381 TI - Modification by pyrethroids and DDT of phosphorylation activities of rat brain sodium channel. AB - The effects of pyrethroids and DDT on the alpha-subunit protein of the rat brain sodium channel were studied by using both native and exogenously added cAMP dependent protein kinases. For this purpose, the sodium channel was partially purified, using the method of Hartshorne and Catterall [J Biol Chem 259: 1667 1675, 1984], and 32P-phosphorylated using [gamma-32P]ATP and exogenously added catalytic subunit of cAMP-dependent protein kinase. By comparing the phosphorylation patterns of the isolated sodium channel to those of the partially purified or unpurified (i.e. intact synaptosomes) preparations, it was concluded that the alpha-subunit of the voltage-sensitive sodium channel protein is the only phosphorylatable protein present at the 260 kD molecular weight range on the sodium dodecyl sulfate-polyacrylamide gel electrophoretogram. Phosphorylation of the alpha-subunit was induced by depolarization, and this process was inhibited by 10(-6) to 10(-10) M 1R-deltamethrin, but not by 1S-deltamethrin, the latter being an inactive enantiomer of the former. DDT produced a similar effect, but only at a higher concentration range. By using lysed synaptosomal membranes, it was possible to study the direct effects of these compounds on the alpha-subunit, which were similar to those produced by depolarization of intact synaptosomes. PMID- 2547383 TI - Calcium mobilization, prostaglandin E2 and alpha 2-adrenoceptor modulation of glucose utilization and insulin secretion in pancreatic islets. AB - alpha 2-Adrenoceptor agonists inhibit glucose-stimulated insulin release and glucose utilization in pancreatic islets. In isolated pancreatic islets of the rat, the Ca2+ channel agonists CGP-28392 and BAY-K-8644 increased insulin release in the presence of clonidine. Neither CGP-28392 nor BAY-K-8644 antagonized the effect of clonidine on glucose utilization. The Ca2+ ionophore, ionomycin, also did not affect glucose utilization in the presence or absence of clonidine. Glucagon partly reversed the effects of clonidine on insulin release, and it potentiated glucose-stimulated insulin release in the absence of clonidine. Glucagon reversed the effects of clonidine on glucose utilization. Amiloride antagonized the effects of clonidine on insulin secretion but did not enhance markedly glucose utilization in the presence or absence of clonidine. Carbamylcholine and arecoline reversed the effects of clonidine on glucose utilization and partly reversed the effects on insulin release in the absence of extracellular Ca2+. Prostaglandin (PG) E2, but not PGF2 alpha, inhibited glucose utilization in a time- and concentration-dependent manner. PGE2 also inhibited glucose-stimulated insulin release. Pertussis toxin blocked both actions of PGE2. The cyclooxygenase inhibitor indomethacin did not affect insulin release or glucose utilization in the presence of clonidine. Thus, elevated intracellular Ca2+ levels antagonize the effects of clonidine on insulin release, whereas other mediators appear to be required to alter glucose utilization. PMID- 2547384 TI - The effect of the plasticizers TBEP (tris-(2-butoxyethyl)-phosphate) and DEHP (di (2-ethylhexyl)-phthalate) on beta-adrenergic ligand binding to alpha 1-acid glycoprotein and mononuclear leukocytes. AB - The plasticizers tris-(2-butoxyethyl)-phosphate (TBEP) and di-(2-ethylhexyl) phthalate (DEHP) and the beta-adrenergic receptor-blockers [3H]-(-) dihydroalprenolol ([3H]-(-)-DHA) and [3H]-(-)-CGP 12177 were tested for their ability to interact with beta-adrenergic binding to alpha 1-acid glycoprotein (AAG) and mononuclear leukocytes (MNL). The IC50-values, obtained by displacement of [3H]-(-)-DHA bound to AAG, were 3.5 nM, 2 microM and 4 microM for TBEP, (-) alprenolol and DEHP, respectively. (+/-)-CGP 12177 had virtually no effect on radioligand binding to AAG. The [3H]-(-)-CGP 12177 binding to MNL consisted of beta-adrenergic receptor binding (Kd = 210 pM) and non-saturable binding. [3H]-( )-DHA was bound to two different classes of binding sites on MNL, the beta adrenergic receptors (Kd = 440 pM) and a secondary class of binding sites (Kd = 64 nM). (+/-)-CGP 12177 displaced about 30% of [3H]-(-)-DHA from MNL with an IC50 value of 190 pm. (-)-ALP displaced about 85% of total bound radioligand and gave a biphasic displacement curve with IC50-values of 320 pM and 690 mM, respectively. TBEP displaced a considerable fraction of [3H]-(-)-CGP 12177 and [3H]-(-)-DHA bound to MNL beta-adrenergic receptors, whereas DEHP had no effect. In contrast, DEHP caused displacement of [3H]-(-)-DHA from the MNL low affinity sites, but was a markedly less potent displacer compared to TBEP. The present study shows that TBEP and DEHP interact with beta-adrenergic transport proteins, non-specific tissue binding sites and beta-adrenergic receptors coupled to adenylate cyclase. Plasticizers may thus affect the biology and pharmacology of the beta-adrenergic signal system. PMID- 2547385 TI - Synergism between chemotactic peptide and platelet-activating factor in stimulating thromboxane B2 and leukotriene B4 biosynthesis in human neutrophils. AB - Formyl-Met-Leu-Phe (FMLP) and platelet-activating factor (PAF) were capable of stimulating thromboxane B2 (TXB2) and leukotriene B4 (LTB4) syntheses in human neutrophils, albeit in a relatively poor degree. A combination of FMLP and PAF, however, was synergistic in stimulating TXB2 and LTB4 syntheses. Phorbol myristate acetate (PMA) appeared to attenuate PAF- but not FMLP-induced arachidonate metabolism. These results suggest that cooperative action of FMLP and PAF on arachidonate release and metabolism does exist and that PMA-mediated protein kinase C activation may regulate FMLP and PAF actions in a different manner. PMID- 2547386 TI - L-6-ketopiperidine-2-carbonyl-L-leucyl-L-proline amide as a novel thyrotropin releasing hormone analogue with improving effects on impaired central nervous systems functions. AB - Psychopharmacological activities of L-6-ketopiperidine-2-carbonyl-L-leucyl-L proline amide (RGH-2202) were compared with those of the parent peptide, thyrotropin releasing hormone (TRH, L-pyroglutamyl-L-histidyl-L-prolinamide, Pyr His-Pro amide) in rodents. RGH-2202 caused qualitatively similar effects to TRH in a variety of tests for the activity on the central nervous system. The compound as well as TRH increased the spontaneous motor activity, enhanced the conditioned avoidance response, and antagonized the avoidance suppression by haloperidol and the sleep-inducing effect of pentorbarbital in mice. It was also efficacious against the deficits of consciousness in mice with concussive head injury and the EEG disturbance in rats with basilar artery occlusion. Besides, it antagonized the cycloheximide-or anoxia-induced amnesia and enhanced the habituation of exploratory activity in mice. Biochemically, it enhanced, like TRH, the turnover of norepinephrine and dopamine in the cerebral cortex, nucleus accumbens and striatum of mice and rats, and increased cyclic GMP levels in the cerebellum of rats. In spite of the global similarity between the pharmacological profiles of RGH-2202 and TRH, there were some intriguing differences between them. RGH-2202 was 2-5 times more effective than TRH in improving the deficits of active avoidance performance and retention in mice, while it was weaker than TRH in modifying the haloperidol-induced catalepsy in mice and enhancing the spinal reflexes in rats. Besides, its thyrotropin releasing activity was about 30 times less potent than that of TRH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547387 TI - Antihypertensive mechanism of alacepril. Effects of its metabolites on the peripheral sympathetic nervous system. AB - To elucidate the antihypertensive mechanisms of alacepril (DU-1219), the drug itself and its metabolites, desacetylalacepril (DU-1227) and captopril, were examined both in vitro and in vivo for their effects on the sympathetic nerve which innervates the peripheral vessels. 1. In isolated perfused mesenteric preparations from spontaneously hypertensive rats (SHR, DU-1227 (10(-6)-10(-5) mol/l) attenuated dose-dependently the increases in perfusion pressure and in norepinephrine (NE) overflow which were induced by electrical stimulation of periarterial sympathetic nerves (15 Hz). Captopril (10(-6)-10(-5) mol/l) caused a similar attenuation, though to a lesser degree, of the perfusion pressure but did not inhibit the increase in NE overflow. 2. The sympatho-inhibitory effect of DU 1227 in the above experiment was shown to be caused by DU-1227 per se, since no captopril was detected in either the perfusate or tissues perfused with DU-1227. 3. In pithed SHR, alacepril (3 mg/kg) caused as potent an inhibitory effect as captopril (3 mg/kg) on the pressor response to the electrical sympathetic nerve stimulation (3 Hz) at an oral dose about half as that of captopril on the molar basis. The effect of alacepril tended to last longer than that of captopril. However, at higher oral dose levels, the inhibitory effect of alacepril (30 mg/kg) was of the similar extent to that of captopril (30 mg/kg). 4. In pithed SHR which had received bilateral nephrectomy 2 to 8 h previously, alacepril (30 mg/kg p.o.) significantly attenuated the vasopressor response induced by electrical stimulation (1-30 Hz) 1 and 3 h after administration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547388 TI - Antisecretory activity of 6-aminoindazole in rats. AB - Administration of 6-aminoindazole (6-AIN) and 5-aminoindazole (5-AIN) to rats depressed gastric acid secretion, both basal and carbachol-stimulated. 6-AIN proved to be more active than 5-AIN in decreasing the stimulated secretory process. The antisecretory activity of 6-AIN appears to be partially an antihistamine effect, since this compound antagonized the stimulatory action of betazole on isolated guinea pig auricle. The antisecretory activity of 6-AIN is probably associated with an antimuscarinic effect, since the molecule decreased carbachol-stimulated gastric acid secretion in rats and depressed neostigmine stimulated motility of duodenum and colon in the anaesthetized cat. It also lessened the hypertonus of isolated guinea-pig trachea caused by pilocarpine. The antisecretory effects of 6-AIN also appear to be associated with myolytic activity, since it decreased the spontaneous motility of duodenum and colon in anaesthetized cats and the spontaneous myoactivity of isolated jejunum of the rabbit. It depressed the contractions of isolated guinea-pig ileum caused by histamine and decreased the spasmogenic effects of barium chloride on isolated guinea-pig gall bladder. These results suggest that 6-AIN probably depresses gastric acid secretion by interfering with both histamine and acetylcholine receptors and with other receptors involved in the secretory process. PMID- 2547389 TI - Very long-acting narcotic antagonists: the 14 beta-p-substituted cinnamoylaminomorphinones and their partial mu agonist codeinone relatives. AB - The biological activity of 14 beta-(p-halo and p-methylcinnamoylamino)-7,8 dihydro-N-cyclopropylmethylnorcodei nones and their corresponding morphinones was investigated. In vitro, the codeinones displayed predominantly mu agonist activity in the 3H-etorphine binding assay and mouse vas deferens preparation. In vivo, in the mouse, the compounds showed weak to inactive antinociception in the tail-flick and hot-plate tets; however, they were potent agonists in the phenylquinone test and moderately weak antagonists in the tail-flick vs morphine test. They also substituted for morphine in withdrawn morphine-dependent rhesus monkeys. When given to non-withdrawn morphine-dependent monkeys, the codeinones precipitated a delayed but long-lasting withdrawal syndrome. They all generalized to codeine in the drug-discrimination test in rhesus monkeys and, in the dose range tested, two compounds were self-administered. This activity is consistent with that of a partial mu agonist. On the other hand, all the morphinones had very long-acting and highly potent mu antagonist properties. The data can be reconciled by assuming that the codeinones are partially metabolized to their respective morphinones. These compounds may be especially useful in the treatment of opioid dependence. PMID- 2547390 TI - Gastric antisecretory and antiulcer/cytoprotective effects of 2-cyano-3 (ethylthio-3-methylthio)-2-propenoic acid methyl ester. AB - AY-28,200 (2-cyano-3-(ethylthio-3-methylthio)-2-propenoic acid methyl ester), a new gastric antisecretory/antiulcer agent, inhibited basal and pentagastrin stimulated gastric acid secretion in the conscious rat (ED50 = 7.6 and 1.9 mg/kg i.g., respectively). For inhibition of basal secretion, peak activity was attained in 5 to 6 h after dosing and was maintained for more than 10 h, with no gastric antisecretory activity occurring at 24 h. The K+ stimulated H+-K+ ATPase activity from rabbit gastric microsomes was inhibited by AY-28,200 (IC50 = 22 mumol/l). AY-28,200 inhibited ethanol-induced gastric lesions, at 3 mg/kg p.o. AY 28,200's cytoprotective effects against ethanol lasted for more than 4 h. AY 28,200 blocked acetylsalicylic acid and stress-induced gastric ulcers but was inactive against indomethacin-induced gastric ulcers. These results suggest that AY-28,200 is a parietal cell proton pump inhibitor with cytoprotective properties, and may produce its cytoprotective effect by stimulating the formation of endogenous prostaglandins. PMID- 2547391 TI - [Malignant mesodermal mixed tumor of the ovary with neuro-ectodermic differentiation]. AB - A striking ovarian tumour was observed in a post-menopausal, 54-year-old woman. Most of the tumour shows histologic features of a malignant mesodermal (mullerian) mixed tumour. However, this tumour harbours a peculiar component suggesting neuroectodermal neoplasia. Histochemical and immunohistochemical studies demonstrate focal pheochromocytomatous (or paraganglionic) and ganglionic differentiations. Several types of cells could be identified: ganglion cells (NSE +, neurofilaments +), pheochromocytes or paraganglion cells (chromogranin +), Schwann cells (S 100 protein +), and neuroblastomatous elements (NSE +). This neoplastic component could be compared with compound tumours such as pheochromocytomaganglioneuroblastoma or gangliocytic paraganglioma. This type of tumour has not been reported in ovary, notably in association with a mesodermal mixed tumour. Such observation prompts us to consider tumours of somatic origin containing germ cell-type elements, with a review of the literature. PMID- 2547392 TI - Hospital practices and breastfeeding duration: a meta-analysis of controlled trials. AB - Controlled clinical trials studying the influence of hospital practices on breastfeeding duration were combined using meta-analysis. Nine studies were selected. Supplementation demonstrated a negative clinical effect on breastfeeding duration that was not significant. Nursing support demonstrated a positive clinical effect on breastfeeding duration that was not statistically significant if there was no telephone follow-up, but was significant if there was telephone follow-up. Early contact revealed a positive clinical effect that was significant. We conclude that early contact and nursing support with telephone follow-up appear as enhancing factors of breastfeeding duration. PMID- 2547393 TI - [Fiber emissions from weathered asbestos cement products. 2. Physical-chemical properties of liberated asbestos fibers]. AB - It could be already shown that fibrous particulates can be emitted from weathered and corroded surfaces of asbestos-cement-(AC)-products and that these fibers increase the total ambient air concentration after having been dispersed in the atmosphere. In this investigation we studied the influence of the polluted atmosphere on physical and chemical properties of asbestos fibers in the corroded surface layer. It was found, that in a part of these asbestos fibers the magnesium was partly leached and that the fibers were strongly contaminated by deposited air pollutants. PMID- 2547394 TI - Critically ill polyneuropathy associated with burns: a case report. AB - A severe sensorimotor peripheral neuropathy is reported in a patient with burns covering 35 per cent of the body surface area, severe sepsis and acute renal failure. It presented as difficulty in wearing the patient from a ventilator. Attention is drawn to the condition of critically ill polyneuropathy which has been recently noted in severe illnesses and sepsis. From a search of the literature we believe this to be the first report of this condition in association with thermal injury. PMID- 2547395 TI - [Genital herpes infection in Chilean female university students]. AB - This article reports on the prevalence of genital herpes infection in a sample of 635 female university students who were seen at the Gynecology Center of the University of Chile Student Medical Service. Viral isolation of genital secretions yielded 18 students (2.8%) with positive results, of whom 28% were asymptomatic shedders. Of the positive cases, 61% were clinically classified as recurrent and 39% as first episodes of genital herpes. Fluorescent monoclonal antibody typing of the isolated strains revealed that 72% were herpes simplex type 2 and 28% were type 1. The results confirm the importance of type 2 virus in genital herpes infections. They also point to asymptomatic shedders as a potential source of infection, and show the importance of a history of genital herpes in the identification of this population. PMID- 2547396 TI - Selective lesion of central dopamine or noradrenaline neuron systems in the neonatal rat: motor behavior and monoamine alterations at adult stage. AB - Different parameters of motor behavior (locomotion, rearing and total activity counts) were studied in the adult rat following neonatal intracisternal 6 hydroxydopamine (6-OHDA, 50 micrograms) treatment combined with noradrenaline (NA) uptake blocker (desipramine) or dopamine (DA) uptake blockers (amfolenic acid or GBR 12909) to obtain selective DA or NA lesions respectively. At 61-65 days of age, selective DA-lesioned animals showed an initial decrease in spontaneous motor behavior at test days 1 and/or 2, while at test days 4 and 5 hyperactivity was observed. However, following amfolenic acid or GBR 12909 pretreatment leading to a selective NA lesion, no difference in spontaneous motor behavior was seen on any of the 5 test days. Determination of regional brain levels of NA and DA confirmed the type of lesion predicted from the various pretreatments with selective uptake blockers. These data suggest that changes in motor behavior in the adult rats, following neonatal 6-OHDA treatment, are specifically related to a DA-denervation, whereas an NA lesion does not seem to influence the spontaneous motor behavior. However, following the selective DA lesion, significant increases of serotonin levels in striatum and cerebellum were observed, while following selective NA lesions an increase of cerebellar NA levels was found concomitant with drastic reductions of NA levels in frontal cortex and spinal cord. PMID- 2547397 TI - Regional hemodynamic abnormalities in overweight men. Focus on alpha-adrenergic vascular responses. AB - Blood pressure, forearm hemodynamic data, and red blood cell (RBC) cation transport measurements were obtained in 27 men with body mass indexes (BMI) greater than 26.6 kg/m2 and 21 men with BMI less than 26.6 kg/m2. Subjects with higher BMI had significantly higher systolic and diastolic pressure measured both noninvasively and invasively, P less than .02-.001. Baseline forearm blood flow (FABF) was substantially higher in the heavier subjects, P less than .02, whereas forearm vascular resistance (FAVR) levels were not different, P greater than .10. Despite the higher baseline forearm blood flow, the high BMI group displayed a greater rise in flow, P less than .01, in response to a regional phentolamine infusion, an index of vascular alpha-tone. Results of multiple regression analysis suggested that overweight was indirectly correlated with vascular alpha tone via a relationship with arterial plasma norepinephrine concentration, a marker of sympathetic drive, and bumetanide-sensitive RBC Na+-K+ cotransport activity. These data indicate that in overweight men (1) higher FABF is not fully counter-balanced by lower FAVR; (2) increased vascular alpha-tone contributes to the altered flow-resistance relationship and thereby to the elevated blood pressure; and (3) greater sympathetic drive and Na+-K+ cotransport activity may explain the elevated vascular alpha-tone. PMID- 2547398 TI - Facilitatory effects of ouabain and digitalis-like substance on adrenergic transmission in hypertension. AB - The purpose of the present study was to examine the role of Na+,K+-ATPase activity in vascular adrenergic transmission of hypertension. In isolated perfused mesenteric vasculatures of spontaneously hypertensive rats (SHR, Okamoto and Aoki strain, seven- to ten-weeks-old) and age-matched Wistar Kyoto rats (WKY), the effects of ouabain, a potent Na+,K+-ATPase inhibitor, or partially purified plasma obtained from salt-induced hypertension on pressor responses and norepinephrine overflow were investigated. Pressor responses and norepinephrine overflow during electrical nerve stimulation were significantly greater in SHR than in WKY. Ouabain increased the stimulation-evoked pressor responses and norepinephrine overflow. This facilitation was more prominent in SHR than in WKY. Partially purified plasma obtained from reduced renal mass-salt hypertensive rats, which had a crossimmunoreactivity with digoxin, also increased the pressor responses and norepinephrine overflow during electrical nerve stimulation, and the effects were greater in SHR than in WKY. These results suggest that Na+,K+ ATPase on vascular adrenergic neurons has an important role in the regulation of neurotransmitter release, and that its activity might be enhanced in SHR. PMID- 2547399 TI - Central mechanisms of hypertension. AB - Central monoaminergic neurons, particularly noradrenergic neurons, are involved in the pathogenesis of hypertension. Dietary NaCl supplementation in NaCl sensitive spontaneously hypertensive rats (SHR-S) elevates blood pressure, increases peripheral sympathetic nervous system activity, and depresses endogenous noradrenaline stores and noradrenaline release in the anterior hypothalamus (AHA). NaCl-resistant spontaneously hypertensive rats (SHR-R) and normotensive Wistar-Kyoto (WKY) rats are resistant to the NaCl-induced alterations in blood pressure and central and peripheral noradrenergic activity, whereas uninephrectomized Sprague-Dawley rats treated with DOCA + NaCl and Dahl-S rats receiving dietary NaCl supplementation develop NaCl-induced increments in blood pressure, but not depressed AHA noradrenaline release. The AHA is a major cardiovascular regulatory region, and depressor responses elicited by pharmacological (alpha 2-adrenoceptor) stimulation of this area are exaggerated in SHR-S fed a high NaCl diet compared to SHR-S fed a basal diet and SHR-R and WKY fed a high or basal NaCl diet. alpha 2-Adrenoceptors in the AHA are increased in number in SHR-S fed a high NaCl diet, presumably reflecting upregulation in response to reduced local noradrenaline release. These findings are consistent with the hypothesis that decreased noradrenergic activity of sympathoinhibitory neurons in the AHA may mediate the exacerbation in hypertension that occurs in SHR-S during dietary NaCl supplementation. PMID- 2547400 TI - Effect of dietary sodium on the Na-K ATPase inhibitor in patients with essential hypertension. AB - To study the circulating humoral factor modifying transmembrane sodium transport, plasma was obtained from 12 patients with essential hypertension (EH) fed a high sodium diet (NaCl 15 to 17 g/d) for seven days and thereafter a low sodium diet (NaCl 2 to 3 g/d) for seven days. Ouabain-sensitive 86Rb+ influx into the red blood cells (RBC) obtained from a healthy subject, and incubated with the plasma obtained during the high sodium diet was significantly lower than that incubated with the plasma obtained during the low sodium diet (3.74 +/- 0.26 v 3.97 +/- 0.30 nmol/10(8) cells, P less than .05). The changes in mean blood pressure from the high to low sodium diet showed a significant positive correlation with the changes in the ouabain-sensitive Rb influx into RBC in the plasma from the high to low sodium diet. These results suggest that a humoral factor modifying the sodium pump might be altered by sodium balance in EH, especially in salt sensitive hypertension. PMID- 2547401 TI - Na,K-ATPase activity in renal tubule cells from Milan hypertensive rats. AB - Several abnormalities of cation transport have been described in the Milan hypertensive rats (MHS). In this study we examined Na,K-ATPase activity in proximal convoluted tubules (PCT) cells and medullary thick ascending limb of Henle cells (TAL) from MHS and from the Milan normotensive rats (MNS). Na,K ATPase activity was determined as 32P-ATP hydrolysis in single tubule segments. Na,K-ATPase activity (pmol Pi/mm t/h) was significantly higher in MHS than MNS both in PCT (903 +/- 227 n = 8 v 506 +/- 285 n = 12) and TAL (4324 +/- 800 n = 5 v 3063 +/- 625 n = 5). Na,K-ATPase dependent respiration was determined in PCT cell from MNS and MHS. Under basal condition Na,K-ATPase dependent respiration (mumol O2/mg protein/h) was higher in MHS than in MNS (24.2 +/- 1.8 n = 5 v 16.1 +/- 0.4 n = 5). When the cells were Na loaded by amphotericin Na,K-ATPase dependent respiration increased significantly more in MHS than MNS (38.4 +/- 1.6 v 26.8 +/- 2.2 n = 4). Thus, Na,K-ATPase activity is higher in renal tubule cells both at normal intracellular Na and after the cells have been Na loaded. The results indicate that regulation of Na homeostasis in renal tubule cell is different in MHS and MNS. PMID- 2547402 TI - [High extracellular concentrations of potassium and rubidium modulate the synaptic transmission in the frog labyrinth]. AB - High extracellular K or Rb levels (20 mM) produce an increase in the resting EPSP and spike frequencies recorded intra cellularly from single fibres of the posterior nerve in the isolated frog labyrinth. The afferent discharge facilitation proved to be inversely related to the fibre's initial resting activity. The K effect is systematically larger than the Rb effect. High sensitive and scarcely sensitive units may be identified with respect to K and Rb action. The present findings suggest that, according to previous models of hair cell functioning, the K and Rb effects are mediated by a raise in intracellular Ca concentration which sustains an increased transmitter release at the cyto neural junction. PMID- 2547403 TI - The influence of the light-dark and wake-sleep cycles on preoptic cAMP concentration in the rat. AB - The concentration of cAMP was measured in the preoptic region of rats sleeping at normal laboratory conditions (12:12 h LD, Ta 22 +/- 0.5 degrees C). The results show that the nucleotide concentration changed both during the circadian LD cycle and during the ultradian wake-sleep cycle. PMID- 2547404 TI - The effect of propranolol on cAMP concentration in the rat preoptic region during the wake-sleep cycle. AB - In control conditions preoptic cAMP concentration during wakefulness was significantly higher than during synchronized sleep. No differences in nucleotide concentration were observed in the cerebral cortex. Propranolol decreases brain cAMP concentration. This change was associated with the suppression of the difference observed between wakefulness and synchronized sleep in the preoptic region. PMID- 2547405 TI - Importance of cholescintigraphy and inferior vena cava flow studies in the differential diagnosis of hepatocellular carcinoma. AB - In order to assess the usefulness of inferior vena cava flow studies and cholescintigraphy complementary to the routine static liver scintigraphy in the differential diagnosis of hepatocellular carcinoma (HCC), we studied 37 patients with a proven diagnosis of HCC and 11 patients with a liver abcess or cyst. The procedure followed was 1) a 99mTc-colloid flow study of the inferior vena cava (IVC) and iliac veins followed by static liver imaging and 2) cholescintigraphy using a dynamic acquisition mode to determine the perfusion as well as the concentration/excretion of the liver and pathological area. The hepatic perfusion index (HPI) was calculated by the "slope" method of Sarper et al.: Radiology 141:179-184 (1981) and the "area" method of Biersack et al.: Fortschritte auf dem Gebiete der Rontgenstrahlen und der Nuklear medizin. Erganzungsband 126:47-52 (1977). The results were compared with data previously collected in patients without liver disease (control) and other liver pathologies. PMID- 2547406 TI - Investigative study of radiopharmaceuticals useful for imaging skeletal muscle injury in experimental animals. AB - An experimental animal model for studying skeletal muscle injury is described. Tc 99m PYP is accumulated on the skeletal muscle injury, but its uptake on the adjacent bone obscures its usefulness in delineating the extent of the muscle injury. In-111 antimyosin accumulates and delineates the extent of the skeletal muscle injury and does not accumulate on the adjacent bone. Hence, In-111 antimyosin is a good radiopharmaceutical for studying the severity and prognosis of skeletal muscle injury. PMID- 2547407 TI - Myocardial beta-adrenoceptor function and regulation in heart failure: implications for therapy. PMID- 2547408 TI - Bradykinin-induced bronchoconstriction: inhibition by nedocromil sodium and sodium cromoglycate. AB - 1. The effects of inhaled nedocromil sodium and sodium cromoglycate on bradykinin induced bronchoconstriction have been studied in a double-blind, placebo controlled study, in eight mild asthmatic subjects. 2. The subjects attended on four occasions. Fifteen minutes after drug pre-treatment a bradykinin challenge was performed. Increasing concentrations were inhaled until a greater than 40% fall in expiratory flow at 30% of vital capacity from a partial flow volume manoeuvre (V p30) was demonstrated. 3. Inhaled bradykinin (0.06-8.0 mg ml-1) caused dose-related bronchoconstriction with the geometric mean cumulative dose causing a 40% fall in V p30 (PD40) of 0.035 (95% CI: 0.02-0.07) mumol, after placebo inhalation, which was similar to that measured before the trial (0.04: 0.02-0.09 mumol). 4. Both nedocromil sodium (4 mg) and sodium cromoglycate (10 mg) gave significant protection (P less than 0.05) against bradykinin-induced bronchoconstriction (PD40 0.37: 0.19-0.72 mumol after nedocromil sodium and 0.22: 0.11-0.49 after sodium cromoglycate). 5. Since bradykinin-induced bronchoconstriction is probably neurally mediated we conclude that both nedocromil sodium and sodium cromoglycate have an action on neural pathways which may be useful in the control of asthma symptoms. PMID- 2547409 TI - Systemic beta-adrenoceptor responses to salbutamol given by metered-dose inhaler alone and with pear shaped spacer attachment: comparison of electrocardiographic, hypokalaemic and haemodynamic effects. AB - 1. Seven normal subjects were given cumulative doubling doses of inhaled salbutamol either by metered-dose inhaler (MDI) alone, or in conjunction with a pear shaped spacer attachment (PSS). Dose increments were made every 20 min from 100 micrograms to 2000 micrograms. 2. Plasma potassium (K), electrocardiographic (ECG) and haemodynamic (HR, SBP and DBP) responses were measured at each dose increment. 3. There were falls in K (as mean and 95% CI) in response to salbutamol (P less than 0.001): 3.70 mmol l-1 (3.46-3.95) to 3.20 mmol l-1 (2.91 3.49) MDI, 3.78 mmol l-1 (3.61-3.95) to 3.18 mmol l-1 (3.06-3.30) PSS. 4. Salbutamol produced marked ECG effects including T wave flattening (P less than 0.001): 0.46 mV (0.24-0.68) to 0.22 mV (0.07-0.37) MDI, 0.50 mV (0.23-0.77) to 0.24 mV (0.07-0.41) PSS; and Q-Tc interval prolongation (P less than 0.001): 0.382 s (0.372-0.392) to 0.409 s (0.397-0.421) MDI, 0.378 s (0.358-0.398) to 0.410 s (0.388-0.432) PSS. U waves occurred in five subjects with MDI and in four with PSS. S-T segment depression was present in two subjects with MDI and in three with PSS. These changes were not however associated with ventricular extrasystoles. There were also significant chronotropic effects (P less than 0.001): 63 beats min-1 (57-70) to 79 beats min-1 (69-89) MDI, 58 beats min-1 (53 63) to 75 beats min-1 (69-81) PSS. 5. Comparison of dose-response curves for MDI alone and with PSS showed no significant differences, for any of the variables measured.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547411 TI - The effect of indomethacin and enalapril on the cutaneous response to bradykinin. AB - The angiotensin converting enzyme inhibitors potentiate the wheal response to intradermal bradykinin. Both converting enzyme inhibitors and bradykinin stimulate prostaglandin synthesis and prostaglandins enhance the cutaneous response to bradykinin. We examined the possibility that the increased wheal response to intradermal bradykinin in the presence of enalapril was due to the effect of prostaglandins. Indomethacin did not inhibit the potentiation by enalapril of the wheal response to bradykinin. PMID- 2547412 TI - Immunocytochemical assay for oestrogen receptor in fine needle aspirates of breast cancer by video image analysis. AB - Assessment of heterogeneity in oestrogen receptor (ER) expression aims to improve prediction of prognosis and treatment assignment in breast cancer. Current assessments are performed manually and are subjective. Automated image analysis as described here objectively quantitates ER in breast cancer nuclei obtained by needle aspiration. ER was visualised by ERICA with diaminobenzidine (DAB) substrate. Various indices of ER positivity were derived from the integrated density and average density measurements of nuclear DAB. Each index was compensated for background staining by non-specific antibody binding and endogenous peroxidase activity. Total nuclear ER content (integrated optical density of stain) was strongly associated with the biopsy ER concentration determined by saturation analysis of radioligand binding (DCC), P less than 0.005. Nuclear ER concentration by image analysis (mean optical density of stain) was not associated with the DCC measurement of ER concentration, P greater than 0.05. This was attributed to technical artefacts of cytocentrifugation. Using threshold values of 5% positive cells and 10 fmol mg-1 concordance of assignment of ER status by image analysis with the DCC assay was 91%, sensitivity was 89% and specificity 100%. It was concluded that image analysis is an appropriate, easy and economic method for determining the nuclear ER status of aspirated cancer cells. Image analysis has the potential to become a powerful diagnostic tool in the assessment of hormone receptor status of breast cancer patients. PMID- 2547410 TI - Metabolic effects of low-dose fluconazole in healthy female users and non-users of oral contraceptives. AB - 1. Azole antifungal agents such as ketoconazole act by inhibiting cytochrome P 450 mediated sterol synthesis in the fungal cell membrane and thus have the potential to interfere with mammalian steroidogenesis. Fluconazole is a novel orally-effective antifungal triazole which has been reported to have more specific effects on the cytochrome P-450 enzymes involved in fungal sterol synthesis. 2. Due to the potential value of systemic antifungal agents in the treatment of infections commonly occurring in women, we assessed the effect of oral fluconazole on the metabolic profile of 18 healthy premenopausal women, 10 of whom were taking combined oral contraceptives (OC). Each woman acted as her own control, being studied both before and 21-28 days after fluconazole therapy (50 mg daily), in the luteal phase of consecutive menstrual cycles. 3. The endocrinological profile included measurement of serum oestradiol, progesterone, testosterone and sex hormone binding globulin (SHBG) concentrations, short tetracosactrin adrenal stimulation test and thyroid function tests. Carbohydrate metabolism was investigated by means of an oral glucose tolerance test with measurement of plasma glucose, insulin and C-peptide concentrations. Serum lipids, lipoproteins and apolipoproteins were analysed on samples taken after an overnight fast. 4. Minor biochemical changes associated with fluconazole treatment included increases in serum thyroxine and testosterone concentrations (but not in women taking OC as well as fluconazole) and in insulin and apolipoprotein B levels (but only in women taking OC as well as fluconazole). In general, these changes were small and of no clinical significance with the values remaining within the laboratory normal range. There were no adverse side effects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547413 TI - In vitro photosensitization of tumour cell enzymes by photofrin II administered in vivo. AB - The ability of injected Photofrin II, a preparation enriched in hydrophobic dihaematoporphyrin ethers and esters, to photosensitize selected mitochondrial and cytosolic enzymes during illumination in vitro was examined. Preparations of R3230AC mammary tumours, obtained at designated times after a single dose of Photofrin II, displayed a time-dependent photosensitivity. Maximum inhibition of mitochondrial enzymes occurred at 24 hours post-treatment, whereas no inhibition of the cytosolic enzyme, pyruvate kinase, was observed over the 168 hour time course. At the selected 24 hour time point, mitochondrial enzyme photosensitisation was found to be drug dose (5.25 mg kg-1 Photofrin II) and light dose dependent, the rank order of inhibition being cytochrome c oxidase greater than F0F1 ATPase greater than succinate dehydrogenase greater than NADH dehydrogenase. We conclude that porphyrin species contained in Photofrin II accumulate in mitochondria of tumour cells in vivo and produce maximum photosensitisation at 24-72 hours after administration to tumour-bearing animals. The time course observed here with Photofrin II is similar to that seen previously with the more heterogenous haematoporphyrin derivative preparation in this in vivo-in vitro model. PMID- 2547414 TI - Microsomal cytochrome P-450-linked monooxygenase systems and lipid composition of human hepatocellular carcinoma. AB - The tissues of hepatocellular carcinoma were operatively resected from six patients. All four components of the systems of microsomal cytochrome P-450 linked monooxygenase of the tissues were investigated and compared to those of normal liver tissue. The concentrations of cytochromes P-450, P-420 and b5 were measured optically and the concentrations of all components except cytochrome P 450 were measured by the Western blotting method followed by immunochemical staining. In microsomes of hepatocellular carcinoma tissues, there was as much cytochrome P-450 and other redox components as in the normal liver tissues, but cytochrome P-450 in liver cancer tissues was unstable and easily converted to cytochrome P-420. The specific activities of NADPH- and NADH-ferricyanide and cytochrome c reductase of each sample were also measured. In the microsomes of the cancer tissues, the specific activities were remarkably reduced compared with those of normal liver tissues. The lipid compositions of the microsomes and the phospholipid/cholesterol ratios (w/w) were 13.1 +/- 3.13 in the cancer tissues and 43.0 +/- 6.74 in normal liver tissues. This difference of the lipid composition elucidates the instability of cytochrome P-450 molecules and the inefficiency of the electron transport of cytochrome P-450-linked monooxygenase systems. PMID- 2547416 TI - Cancer of the breast with nipple involvement. AB - In the absence of gross deformity of the nipple, such as its retraction or Paget's disease, histological examination of this area is often neglected, or at best confined to a cursory look at a single sagittal section. The inadequacy of this approach is illustrated by this study of 33 consecutive cases of carcinoma of the breast treated with mastectomy. Multiple transverse sections showed tumour in 19 nipples (58%) involving one or more levels. Of these, 17 showed non invasive tumour, either ductal or lobular type. Invasive tumour was seen in only two nipples, one of which was metastatic extension from the underlying breast tumour. Paget's cells were seen in two cases. The most significant finding was the eccentric location of tumour in 14 nipples. A single central sagittal section would have detected only five cases involving the centrally situated duct. An inexplicable finding was a preponderance of right nipple with tumour. No statistically significant correlation between nipple involvement and the size, location, multicentricity, type of tumour in the breast and metastases in axillary lymph nodes could be found. It became evident from this study that malignant changes in the nipple occur more commonly than is realised, and that it is also one of the sites of multicentric origin of the tumour. This factor will have to be taken into account in planning conservative therapeutic programmes. PMID- 2547415 TI - Cellular protoonocogenes are infrequently amplified in untreated non-small cell lung cancer. AB - To examine a potential contribution of protooncogene abnormalities other than point-mutational activation of the K-ras protooncogene in the classification of non-small cell lung cancer, amplification of cellular protooncogenes was studied in 47 lung tumour specimens obtained at thoracotomy and in four lung tumour cell lines. The primary tumours included 21 adenocarcinomas, nine large-cell carcinomas, 13 epidermoid carcinomas, one carcinoid and three metastases of primaries outside the lung. The copy numbers per haploid genome of 11 protooncogenes in every tumour sample were determined: H-ras, K-ras, N-ras, c myc, N-myc, L-myc, erbB, mos, myb, ncu (erbB-2) and ral amplifications. The c-myc gene was amplified 5-7-fold in two adenocarcinomas, the H-ras gene 3 5-fold in one adenocarcinoma, while the K-ras and the neu gene were amplified in lung metastases from a colorectal and a breast cancer primary respectively. None of the tumours with an amplified protooncogene simultaneously harboured a mutationally activated K-ras gene. We conclude that amplification of the investigated protooncogenes is a rare event in non-small cell lung cancer. In view of the two c-myc amplifications detected, a systematic study of c-myc expression levels in non-small cell lung cancers appears worthwhile. PMID- 2547417 TI - Peripheral blood mononuclear leukocyte cyclic adenosine monophosphate specific phosphodiesterase activity in childhood atopic dermatitis. AB - Significantly elevated activity of peripheral blood mononuclear cell cAMP specific phosphodiesterase (PDE) activity has been previously reported in adults with atopic dermatitis and in the cord blood of children born to atopic parents. We have been unable to confirm such a significant elevation in children with atopic dermatitis, and we have not found any correlation with dermatitis severity, age or total serum IgE. PMID- 2547418 TI - Isotretinoin differs from other synthetic retinoids in its modulation of human cellular retinoic acid binding protein (CRABP). AB - Isotretinoin differs from acitretin and Ro137410 by its striking sebostatic effect in acne after oral, but not topical, administration. The reason for this is not yet understood. Previous studies indicate that cellular retinoic acid binding protein (CRABP) might be implicated in the action of synthetic retinoids. We, therefore, compared the three retinoids for their ability to increase epidermal CRABP levels after systemic and topical treatment. Oral treatment with acitretin and Ro137410 led to a striking increase of epidermal CRABP (from 2.6 +/ 0.9 to 16.2 +/- 2.9, P less than 0.004 and from 2.5 +/- 1.2 to 21.5 +/- 3.4 pmol/mg protein, P less than 0.004, respectively), while isotretinoin failed to induce a comparable rise (3.2 +/- 1.6 before and 3.7 +/- 0.7 pmol/mg protein after treatment), although it displayed in all patients a striking sebostatic effect. After topical application, the increase of CRABP was comparable for all three compounds. The interaction of isotretinoin with the epidermis seems to be different from other synthetic retinoids only after systemic treatment, a finding that parallels clinical observations. PMID- 2547419 TI - Cutaneous cryptococcosis: treatment with oral fluconazole. AB - A case of cutaneous cryptococcosis is described in an immunocompromised patient. The initial lesion developed on the dorsum of the hand following trauma and was initially thought to be neoplastic. Satellite subcutaneous lesions developed in a 'sporotrichoid' pattern along the forearm. Treatment with oral fluconazole resulted in the complete resolution of the lesions. This is the first published report of the use of fluconazole in the treatment of cutaneous cryptococcosis. PMID- 2547420 TI - Multinucleate cell angiohistiocytoma: a clinicopathological, immunohistochemical and ultrastructural study. AB - The term 'multinucleate cell angiohistiocytoma' was first introduced by Smith and Wilson Jones in 1985. We report the clinicopathological, immunohistological and ultrastructural findings observed in two patients. Multinucleate cell angiohistiocytoma occurs mainly in middle-aged women and is usually located at acral sites, particularly the distal extremities. Grouped, brown-red, slightly elevated, asymptomatic papules slowly develop over several months until further growth ceases. There is no evidence of systemic disease. Histologically, the dermis shows numerous well developed capillaries with prominent endothelia, large bizarre basophilic and often multinucleate cells with a sparse lymphohistiocytic infiltrate. The immunohistological and ultrastructural findings suggest a fibroblastic differentiation of the large multinucleate cells. PMID- 2547421 TI - Enhancement of Candida albicans killing activity of separated human epidermal cells by alpha-melanocyte stimulating hormone. PMID- 2547422 TI - Subcellular composition of platelet density subpopulations prepared using continuous Percoll gradients. AB - Normal human platelets have been separated by density on continuous Percoll gradients and the subcellular composition of platelets of different density has been analysed. The number and concentration of dense granules increased significantly with platelet density, as did the concentrations of the dense granule constituents calcium and serotonin. The amount of serotonin per granule in the low density (LD) platelets was only two-thirds of the corresponding amount in the high density (HD) platelets. Platelets of all densities were able to sequester exogenous serotonin and release it in response to thrombin stimulation with similar efficiencies. The concentrations of the alpha-granule constituents von Willebrand factor and beta-thromboglobulin increased significantly with platelet density but the concentrations of the lysosomal enzyme beta-N acetylglucosaminidase and total sialic acid did not differ significantly in the density subpopulations. The concentrations of the cytosolic enzymes lactate dehydrogenase and glucose-6-phosphate dehydrogenase were slightly higher in the LD population than in the other density subpopulations. The concentration of glycogen showed a marked positive relationship with platelet density and calculations suggested that glycogen was an important determinant of platelet density heterogeneity. The findings of the present study are compatible with recent suggestions that LD platelets may be slightly younger than HD platelets in normal human subjects. PMID- 2547423 TI - Congenital severe protein C deficiency in adults. AB - Two unrelated families are described, of which four adult members were found to be suffering from severe protein C deficiency (less than or equal to 5% of normal plasma level). Newborn deaths were reported in the first family but the second family had no history of neonatal purpura fulminans and infant death. Thrombotic symptoms developed mainly in their early twenties, consisting chiefly of recurrent superficial and deep iliofemoral vein thromboses and pulmonary emboli. Other clinical features included generalized peritonitis due to massive mesenteric vein thrombosis, thrombosis of the cavernus sinus, renal vein thrombosis and priapism. In the second family, five members (all aged approximately 40 years) died of intravascular abdominal thrombosis. Massive thromboembolic episodes were associated with a compensated disseminated intravascular coagulation syndrome as evidenced by high concentrations of D-dimer (mean levels 5000 ng/ml plasma) and by a moderate reduction in platelet count and fibrinogen concentrations. D-dimer levels in noncrisis periods were also raised above normal (mean levels 400 ng/ml). The thrombotic problems of these patients were controlled satisfactorily by long-term administration of low molecular weight heparin alone or in combination with low dose warfarin. PMID- 2547424 TI - Morphologic and morphometric light and electron microscopic studies of the spleen in patients with hereditary spherocytosis and autoimmune haemolytic anaemia. AB - With the aim of contributing to a better understanding of the haemolytic function of the spleen, a morphologic and morphometric study of this organ fixed by arterial perfusion was performed in nine patients with hereditary spherocytosis (HS), three with autoimmune haemolytic anaemia (AHA) and six with Hodgkin's disease without splenic involvement (controls). The spleen weight in HS and AHA (621 +/- 429 g, mean +/- SD) was significantly increased with respect to controls (168 +/- 36 g) (P = 0.003). In HS the red cell retention in the cords of Billroth was significantly increased (203 +/- 68 per 10(4) microns 2) with respect to the cases with AHA (93 +/- 35 per 10(4) microns 2) and to the controls (57 +/- 28 per 10(4) microns 2) (P = 0.004). In HS and AHA the number of macrophages per 10(4) microns 2 of red pulp was significantly increased (5.41 +/- 1.10 and 7.52 +/- 2.91, respectively) with respect to the controls (3.25 +/- 0.58) (P less than 0.003). There was no statistically significant difference between the number of macrophages in HS and AHA. The transmission (TEM) and scanning electron microscopy (SEM) studies demonstrated predominantly red cell retention in the cords of HS spleens, red cell phagocytosis by cordal macrophages in AHA spleens and in a lesser intensity in HS spleens, and phagocytosis of haematic corpuscles by sinus endothelial cells (SEC) in the cases of HS. These quantitative studies allow a better understanding of splenic red cell destruction in haemolytic syndromes. PMID- 2547425 TI - Energy balance in rats given chronic hormone treatment. 1. Effects of long-acting insulin. AB - 1. Sprague-Dawley rats were injected for 16 d with long-acting insulin, and energy balance was calculated using the comparative carcass technique. Two experiments were carried out with females (starting weights 150 and 90 g respectively), and one with males (starting weight 150 g). In a fourth experiment, cytochrome c oxidase (EC 1.9.3.1) activity was measured as an indicator of the capacity for substrate oxidation. 2. Insulin increased weight gain by up to 57% (P less than 0.01 for all studies). Metabolizable energy intake (kJ/d) was also consistently higher in the treated groups, by up to 34% (P less than 0.01 for all studies). The excess weight gained by the insulin-treated rats was predominantly due to fat deposition. 3. Energy expenditure, calculated as the difference between metabolizable intake and carcass energy gain, was expressed on a whole-body basis, or relative to either metabolic body size (kg body weight0.75) or fat-free mass. Insulin consistently raised energy expenditure, regardless of the method of expression, but this change reached statistical significance in only two of the nine comparisons. 4. Cytochrome c oxidase activity was not affected by insulin treatment in either interscapular brown adipose tissue or gastrocnemius muscle. In liver, total enzyme activity (U/tissue) was increased from 2928 (SE 162) in the controls to 3940 (SE 294) in the treated group (P less than 0.02), but specific activity (U/mg protein) was unchanged. 5. It is concluded that, despite causing substantial hyperphagia, insulin treatment only slightly increases energy expenditure in rats. The costs of increased tissue deposition may account for this change. PMID- 2547426 TI - In vitro Na+,K+-ATPase (EC 3.6.1.3)-dependent respiration and protein synthesis in skeletal muscle of pigs fed at three dietary protein levels. AB - 1. Eighteen pigs were offered diets containing 130, 170 or 210 g protein/kg with three barrows and three gifts per diet from 20 to 60 kg live weight. Oxygen consumption, Na+,K+-ATPase (EC 3.6.1.3)-dependent and -independent respiration and protein synthesis were measured in vitro in intercostal and sartorius muscle preparations from these pigs. 2. Increasing dietary protein concentration increased (P less than 0.01) daily gain and dissectible muscle in carcass. 3. O2 consumption and Na+,K+-ATPase-dependent respiration of the intercostal and sartorius muscles increased linearly (P less than 0.01) with increase in dietary protein concentration. The requirement for the support of the transport of Na+ and K+ across the cell membrane in these muscles, on average, accounted for 22 25% of the O2 consumption. 4. Synthesis rate (mg/g per d) of protein in the sartorius muscle increased (P less than 0.05) from 3.05 to 5.07 and increased (P less than 0.1) from 2.57 to 4.06 in the intercostal muscle as dietary protein increased from 130 to 210 g/kg diet. 5. Regression of Na+,K+-ATPase-dependent respiration against protein synthesis in each of intercostal and sartorius muscles showed a linear relation, an attestation of a close link between productive processes and auxiliary energy expenditure. PMID- 2547427 TI - The effect of ouabain on metabolic rate in guinea-pigs: estimation of energy cost of sodium pump activity. AB - 1. The effect of different doses of ouabain, an inhibitor of the sodium pump, or saline (9 g sodium chloride/1; the vehicle) on the metabolic rate of guinea-pigs weighing 500 g was measured by indirect calorimetry for 120 min. 2. Ouabain (0.02 0.07 nmol/g body-weight) decreased the oxygen consumption in a dose-related manner. When higher doses of ouabain (0.10 nmol/g body-weight or greater) were injected the animals were observed to shiver. Ouabain (0.07 nmol/g body-weight) reduced Na+,K+-ATPase (EC 3.6.1.3) activity of liver, kidney and skeletal muscle by 18.0 (SE 6.6), 21.5 (SE 6.0) and 21.9 (SE 6.8)% respectively. An Eadie-Hofstee plot of percentage decrease in O2 consumption v. percentage inhibition divided by dose of ouabain showed that maximal inhibition of O2 consumption was 39.4%. 3. It is concluded that Na pump activity contributed to about 40% of the resting O2 consumption. PMID- 2547428 TI - Riboflavin deficiency, metabolic rate and brown adipose tissue function in sucking and weanling rats. AB - 1. The effects of riboflavin deficiency on growth, whole-body oxygen consumption, cytochrome c oxidase (EC 1.9.3.1) activity and GDP-binding capacity of brown adipose tissue were measured in three groups of rats: sucking pups, weanling rats, and dams. Control groups were weight-matched, pair-fed or fed ad lib. 2. Riboflavin deficiency reduced growth rate and increased the activation coefficient of erythrocyte glutathione reductase (NAD(P)H) (EC 1.6.4.2), as predicted. In sucking pups it also reduced whole-body O2 consumption per unit body-weight, especially after noradrenaline stimulation. In weanling rats, however, it increased O2 consumption both before and after noradrenaline stimulation. 3. Cytochrome c oxidase (EC 1.9.9.1) activity of brown adipose tissue was not consistently affected by riboflavin deficiency. Binding of [3H]GDP to the mitochondria was increased in the deficient weanling rats. 4. Weanling rats therefore, seemed better able to withstand the effects of severe depletion. Their reduced growth and increased non-shivering thermogenesis helped to counteract the unfavourable ratio of riboflavin:other tissue-building materials. The relevance for thermoregulation in riboflavin-deficient children is discussed. PMID- 2547429 TI - Predominant conjugation with glycine of biliary and lumen bile acids in rats fed on pectin. AB - 1. Bile acids were analysed in the bile and lumen samples of rats which received a cholesterol-free or cholesterol-enriched (5 g/kg) diet free from fibre, or containing cellulose or citrus pectin at the level of 100 g/kg. 2. Dietary pectin but not cellulose increased biliary bile acid concentration and excretion. Dietary cholesterol did not affect biliary bile acids quantitatively. 3. Biliary bile acids were almost exclusively conjugated with glycine or taurine in the various experimental situations. The predominant portion of bile acids in rats fed on the cholesterol-free diet was conjugated with taurine when the diet was either free from fibre or contained cellulose; the ratio of bile acids conjugated with glycine: those conjugated with taurine (G:T) was less than 0.2. In contrast, with pectin as a fibre source, the conjugation with glycine increased enormously (G:T increased to approximately 4). Cholesterol enrichment of the diet also increased the glycine conjugation in all groups of rats. Even in this situation, the G:T was highest in rats fed on pectin. 4. Pectin, but not cellulose, increased the bile acid content of the small intestine and caecum, both in rats fed on the cholesterol-free and cholesterol-enriched diets. Cholesterol feeding doubled the bile acid content of the caecum in rats fed on a fibre-free diet or a cellulose diet, but not in those fed on pectin. No such effect of cholesterol was observed in the small intestine, except for the ileal bile acid content in rats fed on cellulose. 5. A considerable portion of the bile acids in the small intestine was deconjugated. The extent of the deconjugation was higher in the ileum than in the jejunum. As in the bile, G:T in rats fed on pectin (3-5.5) were higher than those in the other groups (0.05-1.05) in various situations. Also, cholesterol feeding considerably increased the ratio in all groups of rats. 6. The observed dietary alteration of the partition of bile acids between glycine and taurine may be of physiological significance in regulating bile acid and lipid metabolism in rats. PMID- 2547430 TI - An in vitro method for studying digestion in the pig. 1. Stimulating digestion in the different compartments of the intestine. AB - 1. An in vitro method, using duodenal and ileal digesta and faeces from pigs as inocula for the preparation of three incubation media for simulating the digestive processes in the pig, was proposed. The kinetics of degradation in these three media of three feed samples (pig-grower diet, sugar-beet pulp and wheat bran) with crude protein (nitrogen x 6.25) contents varying from 111 to 196 g/kg, starch contents from 10 to 446 g/kg and dietary fibre contents from 168 to 686 g/kg, were studied. 2. The method was investigated by incubating the feeds in the three media for up to 96 h, and determining the rate and extent of disappearance of feed components. 3. For each feed the 96 h dry matter disappearance was almost equal for all media, although the rate of disappearance varied considerably, with ileal digesta the most potent, particularly for the high-fibre feeds, and duodenal digesta the least. The patterns of disappearance of individual components were similar in all media, with the exception of crude protein which was apparently less degraded in ileal and especially faecal media. 4. Results suggest that a combination of a short (6-12 h) and a long (48-72 h) in vitro incubation could be employed to simulate digestion in the small and large intestine respectively of the pig. PMID- 2547431 TI - An in vitro method for studying digestion in the pig. 2. Comparison with in vivo ileal and faecal digestibilities. AB - 1. An in vitro method involving incubation of feed samples with buffered duodenal digesta for 12 h or with buffered ileal and faecal inocula for 48 h was compared with in vivo ileal or faecal apparent digestibilities in pigs. 2. The five diets investigated had crude protein (nitrogen x 6.25) contents from 164 to 185 g/kg, starch contents from 296 to 463 g/kg and dietary fibre contents from 176 to 347 g/kg. 3. In vitro disappearances with duodenal inocula were correlated (P less than 0.05) with in vivo ileal apparent digestibilities for crude protein, ash, starch, energy and dry matter, but not for dietary fibre. 4. In vitro disappearances with the ileal and faecal inocula were generally correlated (P less than 0.1) with in vivo faecal apparent digestibilities for dietary fibre, energy and dry matter, but not for ash or crude protein. The patterns of degradability of fibre polysaccharide residues in vitro and in vivo were also similar. 5. Results indicate that this in vitro method could be used to predict the availability of starch and crude protein for digestion in the small intestine, and the degradability of dietary fibre, and thus for comparing the nutritive value of pig feeds. PMID- 2547432 TI - Receptor-mediated uptake and 'retroendocytosis' of high-density lipoproteins by cholesterol-loaded human monocyte-derived macrophages: possible role in enhancing reverse cholesterol transport. AB - Human monocyte-derived macrophages (MDM) are cholesterol-loaded, and the rates of uptake, degradation and resecretion of high-density lipoproteins are measured and compared to the rates in control cells. Results show the binding activity of these lipoproteins is upregulated in cholesterol-loaded cells; the bound and internalized lipoproteins are not degraded to any appreciable extent but primarily resecreted as a larger particle. The enhancement of binding activity for high-density lipoproteins is arrested when cycloheximide is added to the medium, suggesting that protein synthesis is involved. Preliminary evidence also indicates that HDL3 (without apoE) after internalisation is converted intracellularly to a larger apoE-containing HDL2-like particles. Thus, MDM appears to possess specific receptors for HDL3 without apoE that may function to facilitate HDL-mediated removal of excess cholesterol from cells. PMID- 2547433 TI - A single and continuous spectrophotometric assay for various lipolytic enzymes, using natural, non-labelled lipid substrates. AB - A rapid, continuous spectrophotometric assay for measuring the amount and activity of several lipolytic enzymes is described. It is based on the metachromatic properties of the cationic dye safranine, and makes use of the fact that an adequate combination of a lipolytic enzyme with one of its substrates leads to a change in the net negative charge at the lipid/water interface, which is monitored by the absorbance change of safranine. Utilizing this method, most lipolytic enzymes can be detected in very low amounts (milliunit or less) in about 1 min without employing radiolabelled lipids or synthetic lipid analogues. Over a wide range of enzyme concentrations, there is a good linearity between the initial hydrolysis rate (determination by the safranine method) and the amount of enzyme. The versatility of the assay is illustrated by examples showing how phospholipase A2, triacylglycerol hydrolase, phospholipase D or phospholipase C (either general or phosphatidylinositol-specific) activities can be detected, either separately or sequentially. Due to its high sensitivity, simplicity, and rapidity, this assay should find its main application in monitoring column effluents during the purification steps of lipolytic enzymes. PMID- 2547435 TI - Characterization of the single-stranded DNA-binding domain of the herpes simplex virus protein ICP8. AB - The DNA-binding protein, ICP8, of herpes simplex virus type 1 (HSV-1) is multifunctional in vivo and binds preferentially to single-stranded DNA (ssDNA) in vitro. To define the ssDNA-binding domain of ICP8, peptides were produced and analyzed. Portions of the ICP8 gene were cloned into the transcription vector pSP64, and RNA was synthesized in vitro. Translation of this RNA in rabbit reticulocyte lysates produced peptides of 29, 35 and 30 kDa, representing amino acid residues 332-564, 571-899 and 900-1196, respectively, of intact ICP8 (128 kDa, 1196 amino acids). These peptides were analyzed by ssDNA-cellulose column chromatography. About 55% of the 29 kDa peptide bound to ssDNA-cellulose columns, and the majority which bound eluted with 1.0 M NaCl. About 5% of the 35 kDa peptide and 12% of the 30 kDa peptide bound and eluted with 0.3 M NaCl. Thus, three regions of ICP8 were associated with ssDNA-binding activity. The ssDNA binding domain of ICP8 was not completely defined, however, because a 95 kDa peptide which included these regions did not bind to or elute from ssDNA cellulose in the same way as intact ICP8. Amino-acid residues 332-564 and 571-899 not only were associated with ssDNA-binding activity but also contain the altered amino acids of four ICP8 molecules which are deficient in DNA binding. PMID- 2547434 TI - Alterations of cholesterol synthesis precursors (7-dehydrocholesterol, 7 dehydrodesmosterol, desmosterol) in dysmyelinating neurological mutant mouse (quaking, shiverer and trembler) in the PNS and the CNS. AB - In brain, levels of cholesterol, desmosterol and 7-dehydrodesmosterol are reduced in shiverer and quaking, but not in trembler 60-day-old dysmyelinating mutant mice. Very interestingly, 7-dehydrocholesterol is not altered in any mutant. The amount of cholesterol is similar in the different normal control mouse strains and in rat. In contrast, levels of precursors are not the same. In sciatic nerve, cholesterol is slightly reduced in shiverer, reduced 2-fold in quaking, and dramatically reduced in trembler (10-fold). 7-Dehydrocholesterol is affected in all mutants. PMID- 2547436 TI - Purification and properties of an endonuclease endogenous to rat-liver nuclei. AB - An endonuclease endogenous to rat-liver nuclei has been purified by a series of chromatographic procedures and finally by isoelectric focusing (IEF) electrophoresis. The nuclease fraction prepared by the IEF electrophoresis (IEF fraction) showed a pI value of 5.7 and migrated as a single band to a molecular weight position of 46,000 on an SDS-polyacrylamide gel. The activity for single stranded DNA was enhanced by 10 mM MgCl2 and/or by 5-15 mM MgCl2 in the presence of 2 mM CaCl2 (an optimum pH, 7.0), but was lowered by CaCl2 alone and inhibited strongly by ZnCl2 or MnCl2. The activity for duplex DNA was rather low, although an optimum condition was 10 mM MgCl2. In fact, even under this condition, the activity was about 40% lower than that for single-stranded DNA. Moreover, the IEF fraction formed single-strand nicks much more rapidly than double-strand cuts in pBR322 DNA, and preferentially produced deoxyadenosine 5'-monophosphate termini in the DNA. In addition, RNAase activity was also detected in this fraction. PMID- 2547437 TI - Mediating effect of calcium in HgCl2 cytotoxicity in sea urchin egg: role of mitochondria in Ca2+-mediated death. PMID- 2547438 TI - Association of thrombin, plasmin, thrombin-antithrombin III complex and plasmin antithrombin III complex with isolated hepatocytes. AB - The interaction of thrombin, plasmin or their antithrombin III complexes with isolated mouse hepatocytes was studied. Plasmin bound to hepatocytes in a concentration-dependent manner with an apparent Kd of 6.4.10(-8) M, attaining equilibrium within 10 min, and the interaction was inhibited by 6-amino-n hexanoic acid. Plasmin treated with diisopropylfluorophosphate (DFP) bound to the cells in similar way as the untreated form of the enzyme. Thrombin bound also to hepatocytes, in a concentration-dependent manner, with a Kd of 5.4.10(-8) M reaching a steady state after 180 min. Thrombin inactivated with DFP, however, was inhibited in its binding to these cells. These data suggest that, whereas the kringle domains of plasmin are responsible for the enzyme-cell interaction, the active center of thrombin may be involved in the binding of this enzyme to hepatocytes. Plasmin-antithrombin III and thrombin-antithrombin III complexes were also associated with hepatocytes in a time-dependent manner, reaching a plateau after 180 min, and the two complexes competed in the interaction. While the interaction of active proteinases plasmin or thrombin with hepatocytes did not result in their internalization, the antithrombin III complexes were taken up by the cells, and thrombin-antithrombin III complex was degraded. These results indicate that hepatocytes may participate in the elimination of proteinase antithrombin III complexes from the plasma, while the association of plasmin and thrombin with hepatocytes could imply distinct biological importance. PMID- 2547439 TI - The Na+/K+-ATPase reaction of human erythrocytes is not near equilibrium. A 31P NMR study. AB - We have addressed the question of whether the Na/K+-ATPase in the human erythrocyte is in a state of near-equilibrium by varying the extracellular ratio of Na+ and K+ and following the cytosolic phosphorylation potential by 31P-NMR and by combined enzymatic colorimetric measurements. There was no correlation at room temperature between the extracellular Na+/K+ ratio and the cytosolic phosphorylation potential measured either by NMR or alternative methods. The cytosolic phosphorylation potential measured by NMR was 4100 +/- 1300 (S.E.) M-1 at an extracellular K+ concentration of 5.9 mM (Na+/K+ ratio of 24.3) and 2800 +/ 700 (S.E.) M-1 at 75 mM extracellular K+ (Na+/K+ ratio of 0.99). The chemically determined phosphorylation potential was 6400 +/- 1200 (S.E.) and 5000 +/- 700 (S.E.) M-1 at 5.9 and 75 mM extracellular K+, respectively. Omission of Ca2+ from the buffer solutions did not affect the results. A consistent finding in this study was that the NMR-determined value of ATP was about 10-20% lower than the value determined enzymatically on perchloric acid extracts. The inorganic phosphate (Pi) was fully NMR visible. PMID- 2547440 TI - Fish oil affects phosphoinositide turnover and thromboxane A metabolism in cultured vascular muscle cells. AB - Fish oil has been reported as having beneficial effects on cardiovascular diseases. Elevated serum lipoproteins, prostaglandins and intracellular free calcium concentrations [( Ca2+]i) of the vasculature and thus the phosphoinositide (PI) turnover may be involved in the pathogenesis of these disorders. Therefore, the effect of fish oil on the potency of both low-density lipoprotein (LDL) and angiotensin II (AII) to stimulate the PI turnover in cultured rat vascular smooth muscle cells (VSMC) has been studied. Furthermore, a possible link between PI turnover activity and thromboxane A2 (TXA2) metabolism in these cells has been investigated. In VSMC cultured for up to 7 weeks with either fish oil or n-3 eicosapentaenoic acid (EPA) a decrease to 5-48% of the LDL induced inositol trisphosphate (IP3) formation (= 100%) was found. A similar range of decreased IP3 synthesis was observed, when AII was used instead of LDL. Both LDL- and AII-stimulated TXA2 synthesis was suppressed concomitantly within the range 34-60%. Blockade of VSMC TXA2 biosynthesis with either indomethacin or TXA2 synthetase blocker (SQ-80338) inhibited LDL-induced formation of IP3 in a dose-dependent manner. Similar results were obtained, when TXA2 receptor coupling antagonists (SQ-27427 or BM-13177) were used. However, blockers of TXA2 synthesis and of TXA2 receptor binding failed to affect AII-induced formation of IP3. PMID- 2547441 TI - Regulation of RAW264 macrophage morphology and spreading: studies with protein kinase C activators, inhibitors and a cyclic AMP analog. AB - Protein kinases C and A probably play important roles in membrane signal transduction. To test the role of protein kinases in macrophage spreading, we have measured cell perimeters in the absence and presence of protein kinase C activators, inhibitors and a cAMP analog. Scanning electron microscopy indicated that macrophages spread extensively in the presence of protein kinase C activators. In contrast, protein kinase C inhibitor and dbcAMP (N6-2'-O-di butyryladenosine 3':5'-cyclic monophosphate AMP) promote a round cell morphology with many surface folds. Quantitative optical microscopy experiments showed that the maximal effects of these reagents were achieved within 30 min. The protein kinase C activators dioctonylglycerol (3 microM), phenylephrine (1 microM), and phorbol myristate acetate (1 micrograms/ml) increased macrophage spreading. Similarly, the calcium ionophore A23187 (1 microgram) increased spreading. In contrast, the protein kinase C inhibitors chlorpromazine (30 microM), sphingosine (10 microM), trifluoroperazine (10 microM), and H-7 (10 microM) significantly reduce macrophage spreading. The analog dibutyryl cAMP (30 microM) abrogates the effects of protein kinase C activators. These data suggest that protein kinase C participates in the regulation of macrophage spreading. Furthermore, the protein kinase A activator dibutyryl cAMP can inhibit the effects of protein kinase C activators. PMID- 2547442 TI - Studies on the binding substances on human erythrocytes for the heat-labile enterotoxin isolated from chicken enterotoxigenic Escherichia coli. AB - To study the predominant binding substance for the heat-labile enterotoxin (LTc) isolated from chicken enterotoxigenic Escherichia coli, competitive binding assays were performed with neuraminidase-treated human type B erythrocytes and 125I-labeled B subunit of LTc (LTc-B). Of all inhibitors used, the ganglioside GM1 was the most effective in inhibiting the binding of 125I-labeled LTc-B to the erythrocytes. The other gangliosides used as inhibitors, gangliosides GD1b, GD1a, GM2, GT1b and GM3, were about 24, 166, 250, 440 and at least 440 times less reactive than ganglioside GM1, respectively. With glycoproteins as inhibitors, on the other hand, hog A + H, porcine thyroglobulin and bovine salivary mucin were over 10(4) times less potent. No inhibition was obtained by other mono-, di- and polysaccharides at the highest concentrations used. These findings suggest that the predominant binding substance on neuraminidase-treated human type B erythrocytes for the LTc-B is ganglioside GM1 and that the combining site of LTc B may be specific for the terminal disaccharide (galactose-N-acetyl-D galactosamine)-linked portion of ganglioside GM1. PMID- 2547443 TI - Protection of cells against membrane damage by haemolytic agents: divalent cations and protons act at the extracellular side of the plasma membrane. AB - The protective effect of Ca2+, Zn2+ and H+ against membrane damage induced by different haemolytic agents has been studied by measuring monovalent cation leakage and haemolysis of erythrocytes, and phosphoryl[3H]choline and adenine nucleotide leakage from Lettre cells prelabelled with [3H]choline. The protective effect of Ca2+ and Zn2+ on erythrocytes damaged by Staphylococcus aureus alpha toxin, Sendai virus or melittin is unaffected by the addition of A23187, even though this ionophore greatly increases the uptake of 45Ca2+ or 65Zn2+. The same result has been found for the protective effect of Zn2+ on Lettre cells damaged by S. aureus alpha-toxin, Sendai virus, melittin or Triton X-100. Leakage of phosphoryl[3H]choline from prelabelled Lettre cells is inhibited if extracellular pH is lowered; lowering the intracellular pH without affecting the extracellular pH, affords little protection. It is concluded that Ca2+, Zn2+ and H+ protect cells against membrane damage induced by haemolytic agents by an action at the extracellular side of the plasma membrane. PMID- 2547444 TI - Characterization and reconstitution of the Na+/H+ antiporter from the plasma membrane of the halotolerant alga Dunaliella. AB - Na+/H+ exchange activity in plasma membrane preparations isolated from the unicellular halotolerant alga, Dunaliella salina, is shown to be competitively inhibited by amiloride or Li+, with Ki values of 25 and 30 microM, respectively. The activity can be followed by either the sodium-dependent change in transvesicular delta pH, as monitored by absorbance changes of Acridine orange, or by the delta pH-dependent uptake of 22Na+ into the intravesicular space. The activity was solubilized, by extraction with Triton, and reconstituted into active proteoliposomes. The activity of the reconstituted proteoliposomes was strongly stimulated by the presence of valinomycin and KCl, suggesting that the exchanger is electrogenic, presumably exchanging more than one proton for each Na+ ion. Partial purification of the Triton-extracted exchanger was obtained by fractionation on a DEAE-cellulose column. PMID- 2547445 TI - Cation sidedness in the phosphorylation step of Na+/K+-ATPase. AB - Na+/K+ -ATPase, reconstituted into phospholipid vesicles, has been used to study the localisation of binding sites of ligands involved in the phosphorylation reaction. Inside-out oriented Na+/K+ -ATPase molecules are the only population in this system, which can be phosphorylated, as the rightside-out oriented as well as the non-incorporated enzyme molecules are inhibited by ouabain. In addition, the right-side-out oriented Na+/K+ -ATPase molecules have their ATP binding site intravesicularly and are thus not accessible to substrate added to the extravesicular medium. Functional binding sites for the following ligands have been demonstrated: (i) Potassium, acting at the extracellular side with high affinity (stimulating the dephosphorylation rate of the E2P conformation) and low affinity (inducing the non-phosphorylating E2K complex). (ii) Potassium, acting at the cytoplasmic side with both high and low affinity. The latter sites are also responsible for the formation of an E2K complex and complete with Na+ for its binding sites. (iii) Sodium at the cytoplasmic side responsible for stimulation of the phosphorylation reaction. (iv) Sodium (and amine buffers) at the extracellular side enhancing the phosphorylation level of Na+/K+ -ATPase where choline chloride has no effect. (v) Magnesium at the cytoplasmic side, stimulating the phosphorylation reaction and inhibiting it above optimal concentrations. PMID- 2547446 TI - The effect of ferricyanide with iodoacetate in calcium-free solution on passive cation permeability in human red blood cells: comparison with the Gardos-effect and with the influence of PCMBS on passive cation permeability. AB - Freshly prepared human red blood cells incubated with 5 mM ferricyanide, 0.2 mM iodoacetate and 2 mM adenosine in the presence of 5 mM EGTA demonstrate comparable increases in Na+ and K+ permeability (ferricyanide effect). This effect is unrelated to the Ca2+-activated K+ channel (Gardos effect) since influx of Ca2+ from outside the cell is excluded. Also this effect is different from the non-specific Na+ and K+ permeability change elicited by PCMBS. These differences become obvious by using various reagents. For example, A23187 and quinidine exert opposite effects in Gardos and ferricyanide experiments, where A23187 and atebrin react oppositely in the latter and in PCMBS experiments. The ferricyanide effect described here does not involve formation of nonspecific channels. The change in Na+ permeability separately from K+ permeability under certain circumstances suggests a more specific effect. PMID- 2547447 TI - Interaction between phosphatidylserine and the isolated cytoskeleton of human blood platelets. AB - Binding experiments were performed to demonstrate a direct interaction between cytoskeletons from human blood platelets and phosphatidylserine. A centrifugation technique using radiolabeled phosphatidylserine-vesicles and Triton X-100 insoluble residues from unstimulated human platelets was used to assess the binding. Interaction between cytoskeleton and phospholipid is demonstrated to be specific for phosphatidylserine. No binding was observed for phosphatidylcholine. The binding of phosphatidylserine was saturable and dependent on the concentration of cytoskeleton used. The interaction between phosphatidylserine and the cytoskeleton appeared to be completely reversible. The existence of a reversible and specific interaction between phosphatidylserine and the cytoskeleton of unstimulated platelets would suggest a role for the cytoskeleton in the maintenance of the asymmetric distribution of this lipid in the plasma membrane. We have previously shown (Comfurius et al. (1985) Biochim. Biophys. Acta 815, 143-148) that in activated platelets a strong correlation exists between degradation of platelet cytoskeletal proteins by the endogenous calcium dependent proteinase (calpain) and exposure of phosphatidylserine at their outer surface. Nevertheless, hydrolysis of the isolated cytoskeleton by calpain did not result in a change in the parameters of the binding between phosphatidylserine and cytoskeleton. Also, sulfhydryl oxidation of the cytoskeleton by diamide did not affect its binding properties for phosphatidylserine, in spite of the fact that diamide treatment of platelets results in exposure of phosphatidylserine at the outer surface. Exposition of phosphatidylserine upon activation of platelets cannot be directly ascribed to a change in affinity or number of binding sites of the modified cytoskeleton as measured in model systems. However, it cannot be excluded that topological rearrangements of the cytoskeleton as occur within the cell during platelet activation lead to a decreased contact between cytoskeleton and lipid, irrespective of the binding parameters. PMID- 2547448 TI - Minimum enzyme unit for Na+/K+-ATPase is the alpha beta-protomer. Determination by low-angle laser light scattering photometry coupled with high-performance gel chromatography for substantially simultaneous measurement of ATPase activity and molecular weight. AB - The oligomeric state of canine renal NA+/K+ -ATPase solubilized by octaethylene glycol n-dodecyl ether (C12E8) was studied by means of low-angle laser light scattering photometry coupled with high-performance gel chromatography (HPGC). At around 0 degree C the solubilized enzyme was separated into the (alpha beta)2 diprotomeric and alpha beta-protomeric protein components with Mr values of 302,000 +/- 10,000 and 156,000 +/- 4,000, respectively, in approximately equal quantities. As the temperature of chromatography was increased toward 20 degrees C, the two protein components converged into a single major component. The Mr of this component depended on the monovalent cation included in the elution buffer, and was 255,000 or 300,000 in the presence of 0.1 M NaCl or 0.1 M KCl, respectively. A computer simulation technique showed that the solubilized enzyme was in a dissociation-association equilibrium of 2 protomers = diprotomer at 20 degrees C, and the difference in apparent Mr of the solubilized enzyme between the two species of monovalent cation was interpreted by an association constant (Ka) in the presence of 0.1 M KCl that was about 50-fold larger than in the presence of 0.1 M NaCl. In order to measure ATPase activity and Mr of the solubilized enzyme simultaneously, a TSKgel G3000SW column had been equilibrated and was eluted with an elution buffer containing 0.30 mg/ml C12E8 and 60 microgram/ml phosphatidylserine (bovine brain) as well as the ligands necessary for the enzyme to exhibit the activity at pH 7.0 and 20 degrees C. The solubilized enzyme was always eluted as a single protein component irrespective of the the amount of the protein applied to the column, ranging between 240 and 10 microgram. The Mr of the protein component, however, decreased from 214,000 and 158,000 with the decrease of the protein amount. The specific ATPase activity, however, remained constant at a level of 64 +/- 4% of that of the membrane-bound enzyme even in the range of protein concentration sufficiently low as to allow the enzyme to exist only in the protomeric form. Thus, the alpha beta protomer is concluded to be the minimum functional unit for the ATPase activity. The value of Ka obtained from the concentration-dependent dissociation curve was 5 . 10(5) M-1 for the enzyme turning over, and 1.1 . 10(7) M-1 for the enzyme inhibited with ouabain. It was discussed, based on the values of Ka obtained, that the enzyme would exist as the diprotomer or the higher oligomer in the membrane. PMID- 2547449 TI - Resistance of cultured hepatoma cells to copper toxicity. Purification and characterization of the hepatoma metallothionein. AB - The mechanism of resistance to the toxic effects of copper was investigated using a series of copper-resistant hepatoma cell lines maintained in copper-enriched medium. Gel electrophoresis of carboxyamidated cell extracts demonstrated the presence of a pair of low molecular mass cysteine-rich proteins in wild-type and resistant cell lines. These proteins were purified to homogeneity and contained approx. 60% of the total cellular copper. Comparisons of molecular masses, pI values and amino-acid compositions for the purified hepatoma proteins with authentic rat liver metallothionein, as well as cross-reactivity with anti-rat metallothionein antibody, confirmed that the cysteine-rich hepatoma proteins were metallothioneins. The cellular concentration of these hepatoma copper metallothioneins was proportional to both the level of metal resistance and the amount of copper accumulated by individual cell lines. Further, resistant cells removed from copper-enriched medium for 6-12 months, yet maintaining their level of resistance, showed only a slight decrease in metallothionein concentration. Thus it is proposed that the level of resistance to metal toxicity is mediated by the concentration of copper-metallothionein. It is also suggested that the steady state level of copper metallothionein is controlled by the degree of metal exposure. PMID- 2547450 TI - The displacement of copper by iron at the specific binding sites of ovotransferrin. AB - We have examined the kinetics and mechanism by which iron can displace copper at the specific metal-binding sites of ovotransferrin. Fe2+ was added to Cu2+ ovotransferrin-CO3(2-) in the presence of NaHCO3 and ambient O2. The reaction has been followed by standard and stopped-flow spectrophotometry, EPR spectroscopy and analysis of chromogen-reactive Fe2+. The reaction is best described as triphasic. An initial jump in absorbance takes place in the first 2 s. In the next minute there is a further increase in absorbance and shift in the spectral maximum from 440 to 446 nm. The third phase is complex. The bulk of the spectrophotometric change, a decrease in absorbance with a shift to a maximum of 453 nm, lasts approx. 3 min. Minor spectral and EPR changes, however, take place over the next several hours. Chromogenic analysis of Fe2+ indicates that approx. 1 min is required to oxidize the Fe2+. EPR spectra reveal the formation of an Fe3+-ovotransferrin complex within the first 20 s; however, this lacks the characteristic doublet of specific Fe3+-ovotransferrin-CO3(2-). The simultaneous presence of specific Cu2+-ovotransferrin-CO3(2-) and Fe3+-ovotransferrin-CO3(2-) signals suggests a period in which the protein specifically binds both metal ions perhaps resulting from a differential reactivity of the two metal-binding sites. The addition of Cu(NO3)2 to Fe3+-ovotransferrin-CO3(2-) resulted in a complex with specific Fe3+ and non-specific Cu2+. The EPR spectrum of this complex and the final product of our displacement reaction were virtually identical. Distinct parallels in reaction of Cu2+-ovotransferrin-CO3(2-) with Fe(NH4)2(SO4)2, Fe(NO3)3 and Fe3+-nitrilotriacetic acid were observed. A reaction sequence involving the binding and oxidation of non-specific Fe2+ followed by Cu2+ displacement by Fe3+ at the specific sites and binding of non-specific Cu2+ is suggested. PMID- 2547451 TI - Spin trapping of lipid-derived radicals in liposomes. AB - Electron-spin resonance-spin trapping has been used to detect lipid-derived radicals in liposomes. Using the lipid-soluble spin trap 2-methyl-nitrosopropane (MNP), we have detected both the lipid and hydrogen-atom spin adducts in liposomes composed of a fully saturated phospholipid (dimyristoylphosphatidylcholine, DMPC) with various mol fractions of unsaturated phospholipid (1-palmitoyl-2-arachidonoylphosphatidylcholine, PAPC) or fatty acid (arachidonic acid, AA). The lipid-derived spin adduct formed during autoxidation of liposomes was separated by thin-layer chromatography and found to co-migrate with the product(s) formed by direct addition of MNP to the corresponding unsaturated lipid or fatty acid. Both the MNP-PAPC and MNP-AA spin adducts showed some restriction of rotational motion when in the liposome bilayer (rotational correlation times 0.72 and 0.69.10(-9) s, respectively), and nitrogen hyperfine coupling constants (14.94-14.96 G) consistent with a hydrophobic localization. Radical versus non-radical mechanisms of spin adduct formation during liposome autoxidation were separated using alpha-tocopherol as a radical scavenger. The utility of nitroso spin traps in trapping of radicals in liposomes is discussed. PMID- 2547452 TI - Thermodynamic bookkeeping: a reinvestigation of proton and dicarboxylic acid binding to aspartate aminotransferase. AB - A reinvestigation of the effects of pH and salt concentration on the proton and dicarboxylic acid dissociation constants of pig heart aspartate aminotransferase shows that both anions and cations were involved concomitantly, both as stoichiometric reactants and bioenergetically. Equations are presented which can be used experimentally, to determine the numbers of salt ions (their thermodynamic stoichiometries) involved in biochemical equilibria such as proton and ligand dissociations from macromolecules. These equations were used to reinvestigate the effects of salts on the chromophoric pKa of the enzyme prosthetic group, the interaction of the enzyme with dicarboxylic acids, and the overall equilibrium for the transamination half-reaction. PMID- 2547453 TI - Proteolytic cleavage of pyridoxal kinase into two structural domains. AB - Chymotryptic digestion of sheep brain pyridoxal kinase, a dimer of identical subunits each of 40 kDa, yields 2 fragments of 24 and 16 kDa with concomitant loss of catalytic activity. These fragments were separated by chromatographic techniques and analyzed for interaction with the ATP analogue, trinitrophenyl ATP, using fluorescence spectroscopy. The absorption and fluorescence properties of trinitrophenyl-ATP bound to the fragment of 24 kDa (emission maximum, 540 nm, emission anisotropy at 460 nm, 0.30, and fluorescence lifetime, gamma = lns) are indistinguishable from those of the ATP analogue bound to the native enzyme. The fragment of 16 kDa does not bind trinitrophenyl-ATP. The results are consistent with the hypothesis that monomeric pyridoxal kinase is folded into 2 domains connected by a single polypeptide chain sensitive to proteolytic cleavage. PMID- 2547454 TI - [Study of nuclear poly(ADP-ribose)polymerase and DNA-topoisomerase II of brain cells during postnatal development of rats]. AB - The nuclear poly(ADP-ribose)polymerase activity of neuronal and glial cells during postnatal development of rats was studied. It was shown that the poly(ADP ribose)polymerase activity of nuclei and nuclear matrix of neuronal cells during postnatal development of rats is increased, whereas the polymerase activity of glial cell nuclei and nuclear matrix in newborn and adult rats is higher than in 14-day-old animals. The DNA-topoisomerase II activity of neuronal nuclear matrix during the postnatal development of rats does not change, whereas the topoisomerase activity of glial nuclear matrix decreases but is always higher than the DNA-topoisomerase II activity of neuronal cell matrix during the postnatal development of rats. It is suggested that ADP-ribosylation in the nuclear matrix of neuronal cells causes the inhibition of the DNA-topoisomerase II activity of nuclear matrix. PMID- 2547455 TI - [Specific splitting of the alpha-A-globin gene by molecular forms of DNA topoisomerase type I from calf thymus]. AB - The specificity of splitting of the cloned alpha A-globin gene from chicken erythrocytes induced by three topoisomerases I differing in molecular masses was demonstrated. The localization and relative number of topoisomerase breaks in the alpha A-globin gene vary in different topoisomerase I forms. PMID- 2547456 TI - [Estimation of a direct regulatory effect of estrogens on hepatocytes, evaluated according to the change in the level of free estrogen-binding protein in primary cell culture]. AB - Male rat hepatocyte cultures have been obtained. The culturing hepatocytes had a stable level of some morphological and functional properties. A high level of estrogen receptors (ER) and E2-sensitive unusual estrogen-binding protein (UEBP) was found. A direct dose-dependent inhibiting effect of physiological (10(-10) 10(-7) M) concentrations of E2 on UEBP content was established in cell cultures incubated with the hormone for 3 days. Hexestrol (but not cholesterol) was found to exert a direct inhibiting effect. The inhibiting effect of E2 (10(-9) M) was observed after a 24 hour incubation of hepatocytes with the hormone but was less pronounced. In this case a significant increase in UEBP concentration in hepatocytes was noted 72 hours after the removal of E2. It is concluded that physiological concentrations of E2 can exert a direct regulatory influence on certain functions of culturing hepatocytes acting via hepatocyte ER. PMID- 2547457 TI - [The enzymology of visual reception: phosphodiesterase cascade of signal amplification]. AB - According to present-day concepts an important and, presumably, a key role in signal transmission in photoreceptor cells is ascribed to a system containing the photosensitive protein rhodopsin, GTP-binding protein transducin and cyclic GMP phosphodiesterase which in many features is similar to the adenylate cyclase system from other eukaryotic cells. The experimental and literary data concerning the already established and hypothetical mechanisms of transmission, enhancement and switch-off of the signal in the rhodopsin----transducin----phosphodiesterase chain are reviewed. PMID- 2547458 TI - [Study on the effect of some group-specific agents on clostridiopeptidase]. AB - The interaction of clostridiopeptidase of Clostridium histolyticum with EDC, TNM and MA, the specific reagents for COOH-groups, tyrosine and lysine residues was studied. It was shown that at pH 6.0 EDC inactivates the enzyme. The inactivation process follows the pseudo-first order kinetics and is described by a second order rate constant equal to 1 M-1 min-1. The synthetic substrate does not prevent, in practical terms, the enzyme inactivation by EDC. At pH 8.0 TNM modifies about 19 tyrosine residues in the clostridiopeptidase molecule which is accompanied by marked inhibition of the enzyme activity (down to 70-90%). In this case, the inactivation process is not described by simple pseudo-first order kinetics but is characterized by two steps (fast and slow) with second order rate constants of approximately 14 and 3.5 M-1 min-1, respectively. The synthetic substrate partly prevents the inactivation of the enzyme by TNM and protects 11 tyrosine residues. The MA-induced incorporation of 13 +/- 3 maleyl groups into the clostridiopeptidase molecule in partially prevented by the synthetic substrate with protects the enzyme against inactivation. The data obtained suggest that lysine residues are seemingly included into the active center of clostridiopeptidase, whereas tyrosine residues provide for the maintenance of active conformation of the enzyme. PMID- 2547459 TI - Down-regulation of testicular aromatization in the horse. AB - A single i.m. injection of testosterone (750 mg of testosterone bexahydrobenzoate) or i.v. injection of human chorionic gonadotrophin (hCG) (10,000 IU) was given to geldings and stallions. Levels of unconjugated and conjugated (after solvolysis) androgens and estrogens were measured in blood and urine samples taken daily from the day of injection (D0) to the tenth day post injection (D10). In the stallion, both treatments resulted in a sharp increase of plasma estrogens, which peaked one day before the androgen levels. Our results confirmed the testicular localization of a potent aromatase, which is able to aromatize androgens from endogenous as well as exogenous origin into conjugated estrogens. The very similar patterns of estrogen increase following testosterone or hCG administration suggest that the estrogen rise induced by hCG results at least partly from increased availability of testosterone. The abrupt drop in plasma estrogen levels cannot be explained by a lack of substrate, since two successive androgen injections did not succeed in maintaining the high estrogen levels. Since estrogens were unable to inhibit the aromatase activity in vitro, the drop in estrogen levels suggests a down-regulation of the aromatase synthesis. PMID- 2547461 TI - Regulation of myometrial beta 2-adrenergic receptors by progesterone and estradiol-17 beta in late pregnant rats. AB - Rat myometrium exhibited a marked decrease in the concentration of beta 2 adrenergic receptors immediately before parturition, i.e., in the last 6 h of pregnancy. This phenomenon continued until the withdrawal of myometrial progesterone (-94% from Day 18 of pregnancy to term) and coincided with the sharp increase (+200%) of the myometrial concentration of estradiol. A linear positive correlation was found (r2 = 0.645) between the concentration of beta 2-adrenergic receptors and the log ratio of myometrial concentration of progesterone/myometrial concentration of estradiol (P/E2), suggesting a modulation of beta 2-adrenergic receptors by steroids. In rats with estrogen dominated uteri (intact of ovariectomized late pregnant rats injected with estradiol), there was no change either in concentration or affinity of beta 2 adrenergic receptors relative to untreated control pregnant rats. In contrast, rats with progesterone-dominated uteri (intact or ovariectomized late pregnant rats treated with progesterone or ovariectomized rats) have an increased number of beta 2-adrenergic receptors, with a decreased affinity of these receptors compared to untreated control pregnant rats or to estrogen-treated rats. These results suggest that progesterone regulates the number of beta 2-adrenergic receptors in myometrium of late pregnant rats. The mechanisms by which progesterone exerts this regulation remains to be elucidated. PMID- 2547460 TI - Human chorionic gonadotropin affects tissue levels of progesterone and cyclic adenosine 3',5'-monophosphate in the metestrus rat uterus in vitro. AB - The effect of human chorionic gonadotropin (hCG) on in vitro progesterone (P) level in the uterus was investigated by using short-term incubations of uterine tissue taken from 4-day cyclic rats at different stages of the estrous cycle. Control incubations resulted in a decrease of endogenous P content in uteri removed from rats in proestrus (PRO), estrus (EST), and metestrus (MET): -25% (n = 6), -60% (n = 6), and -45% (n = 8), respectively. The amount of P found after incubation of MET tissue in the presence of hCG was significantly higher (p less than 0.001) than that found after control incubations. The hCG effect was dose dependent and was not observed with PRO or EST tissue. Although the mean P level found in MET tissue after incubation with 10 IU/ml hCG was not significantly different from the mean level found in unincubated tissue (1562 +/- 341 vs. 1470 +/- 174 pg/mg protein, n = 8), an obvious synthesis was observed in two experiments. It thus seems likely that observed hCG effect would involve a de novo P synthesis rather than a decrease of P catabolism. Furthermore, hCG induced a dose-dependent increase of cyclic adenosine 3', 5'-monophosphate (cAMP) uterine levels in MET tissue. N,O'-dibutyryl cyclic AMP [Bu)2 cAMP) at 5 mM induced a significative increase (p less than 0.01) of P uterine level in EST and MET tissue compared to control incubations, but had no effect on PRO tissue. Our results suggest the progressive maturation throughout the rat estrous cycle of a luteinizing hormone/hCG- and cAMP-dependent process able to regulate uterine P content.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547463 TI - Alteration of the spermatozoal glycocalyx and its effect on duration of fertility in the fowl (Gallus domesticus). AB - The hypothesis that sialic acid has a role in spermatozoal sequestration within the hen's oviduct was tested by treating spermatozoa with Clostridium perfringens neuraminidase. Spermatozoal content of sialic acid ranged from 94 to 135 micrograms per 10(9) spermatozoa (n = 12 roosters). Spermatozoa contained 80% of total seminal sialic acid (coefficient of variation = 4.6%). Spermatozoal sialic acid content was reduced by 18% when 10(9) spermatozoa were incubated at pH 6.5 with 10 IU neuraminidase activity (Type V, Sigma Chemical Co.). Such treatment had no effect on spermatozoal viability as evidenced by ethidium bromide uptake. However, treatment of spermatozoa with neuraminidase prior to intravaginal insemination reduced fertility by 24 percentage units (p less than 0.001). In contrast, when similarly treated spermatozoa were deposited in the magnum via laparatomy, fertility was not affected (p greater than 0.05). The preceding work was done with neuraminidase prepared by salt fractionation (Type V, Sigma Chemical Co.). Type V neuraminidase was absorbed to diethylaminoethyl-Sephacel and then eluted with a stepwise KCl gradient. Treatment of spermatozoa with this preparation of neuraminidase (10 IU/10(9) spermatozoa) prior to intravaginal insemination reduced fertility by 19 percentage units (p less than 0.001). Decreased fertility could not be attributed to contamination of neuraminidase preparation with proteolytic activity. We conclude that spermatozoal sialic acid has a role in spermatozoal sequestration within the hen's utero-vaginal glands. PMID- 2547462 TI - In vitro effect of insulin and insulin-like growth factor-I on cell multiplication and adrenocorticotropin responsiveness of fetal adrenal cells. AB - The present study examined the effects of both insulin and insulin-like growth factor-I (IGF-I) on cell division and specific functions of cultured adrenocortical cells from 100- to 122-day-old ovine fetuses. When culture was performed in a serum-free medium containing transferrin and ascorbic acid, the number of cells increased only slightly (1.2-fold) over a 4-day period. Addition of insulin or IGF-I in the culture medium enhanced the number of cells counted on Day 5. The effect of both peptides was dose-dependent, but 10 ng/ml IGF-I was as potent as 10 micrograms/ml insulin. The acute cyclic adenosine 3',5' monophosphate (cAMP) and steroidogenic responses to adrenocorticotropin (ACTH1 24) decreased in fetal cells cultured in the absence of insulin or ACTH. Insulin at micromolar concentrations not only prevented this decrease but enhanced the acute ACTH1-24-induced cAMP output on Day 5 over that observed on Day 2. Treatment of fetal cells for 4 days with increasing concentrations of insulin or IGF-I enhanced the acute cAMP and steroidogenic responses (3- to 4-fold) to ACTH1 24 over that of control cells. The ED50 of IGF-I was about 3 ng/ml (congruent to 0.4 nM) whereas that of insulin was about 10 ng/ml (1.7 nM). However, a second plateau was apparent at concentrations of insulin above 1 microgram/ml. The acute cholera toxin stimulation of cAMP production of cells cultured in the absence of insulin or ACTH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547464 TI - Buffering the stomach content enhances the absorption of diflunisal in man. AB - Diflunisal, a lipophilic salicylate, is absorbed more slowly in healthy volunteers than aspirin. In this paper we report on attempts to influence diflunisal absorption by buffering the gastric milieu. Sodium bicarbonate given together and 30 min after diflunisal tablets significantly (p less than 0.05) shortened the time to reach peak plasma concentration (tmax greater than 15 per cent), raised maximum plasma concentration slightly (Cmax 6 per cent) and increased the area under the plasma concentration-time curve (AUC greater than 8 per cent). Other pharmacokinetic parameters, including terminal half-life and renal elimination of the compound, were not considerably influenced. These findings indicate that the absorption of diflunisal was enhanced by increased gastric pH, presumably a result of an increased solubility of diflunisal in the stomach together with faster transport into the small intestine. In one volunteer, after intravenous administration diflunisal plasma concentrations declined in a triphasic manner with a terminal half-life of 12.8 h. The volume of distribution was approximately 10 per cent of body weight. Based on the ratio of AUC after equivalent i.v. and oral diflunisal doses, the absolute bioavailability was 89.5 per cent. PMID- 2547465 TI - Pharmacokinetics of lisinopril (IV/PO) in healthy volunteers. AB - When three intravenous doses of lisinopril were administered to healthy volunteers, area under the curve (to infinity) vs dose was linear with a positive intercept. Subtracting area under the extrapolated terminal phase of the serum profile from zero to infinity retained the linear relationship, but shifted the regression line to a zero intercept. It is postulated that the terminal phase reflects binding of drug to angiotensin-converting enzyme (ACE). The half-life for the terminal phase (approximately 40 h) was not predictive of steady-state parameters when ten daily doses (q24h) of lisinopril were administered orally to healthy volunteers. The mean effective half-life for accumulation was 12.6 h. The mean accumulation ratio was 1.38. Steady state was attained after the second daily dose. The observations in these studies with lisinopril are similar to those reported for enalaprilat, the active metabolite of the ACE inhibitor, enalapril maleate. PMID- 2547466 TI - Stimulation of collagenolytic enzyme release from cultured bone cells of normal and osteopetrotic (mi/mi) mice by parathyroid hormone and lipopolysaccharide. AB - Collagenolytic enzyme release from bone cells was studied using cultured calvarial cells which are capable of degrading calcified and noncalcified collagen (cells from normal mice) or only noncalcified collagen (cells from osteopetrotic (mi/mi) mice). Treatment of cells from either normal or mi/mi mice with parathyroid hormone (PTH) or lipopolysaccharide (LPS) resulted in the appearance of latent collagenolytic enzyme activity in the medium. Chromatography of media from cells from normal mice treated with PTH on lysine-Sepharose resulted in the separation of latent collagenase and latent gelatinase. Further characterization of the enzymes showed that they were similar to those previously isolated from media of calvaria cultured with heparin. Collagenase activity of media of cells from normal or mi/mi mice treated with PTH or LPS yielded identical elution patterns upon chromatography on lysine-Sepharose. These results show that the mi allele has no effect on regulation of latent collagenolytic enzyme release. The results show that the mi allele has no effect on regulation of latent collagenolytic enzyme release. The data also suggest that previously described differences between PTH- and LPS-stimulated collagen degradation in cultured calvaria are due to factors other than differences in the ability of these agents to stimulate the release of collagenolytic enzymes. PMID- 2547467 TI - Treatment of amiodarone induced hyperthyroidism with potassium perchlorate and methimazole during amiodarone treatment. AB - To exploit the antiarrhythmic effect of amiodarone when patients develop the side effect of thyrotoxicosis three patients with hyperthyroidism induced by amiodarone were given simultaneously 1 g potassium perchlorate a day for 40 days and a starting dose of 40 mg methimazole a day while they continued to take amiodarone. As hyperthyroidism might have recurred after potassium perchlorate treatment was stopped the dose of methimazole was not reduced until biochemical hypothyroidism (raised thyroid stimulating hormone concentrations) was achieved. The patients became euthyroid (free triiodothyronine concentration returned to normal values) in two to five weeks and hypothyroid in 10 to 14 weeks. One patient became euthyroid while taking 5 mg methimazole a day and 600 mg amiodarone weekly; the two others required substitution treatment with thyroxine sodium while taking 5 mg methimazole or 50 mg propylthiouracil (because of an allergic reaction to methimazole) and 2100 or 1400 mg amiodarone weekly. Hyperthyroidism induced by amiodarone may be treated with potassium perchlorate and methimazole given simultaneously while treatment with amiodarone is continued. PMID- 2547469 TI - Near death experience and the NMDA receptor. PMID- 2547468 TI - Efficacy and long term effects of antenatal prophylaxis with anti-D immunoglobulin. AB - OBJECTIVE: To measure the safety and efficacy of antenatal treatment with anti-D immunoglobulin. DESIGN: Open study with historical controls. SETTING: Multicentre study in 17 hospitals in West Yorkshire. PATIENTS: 1238 Rh negative women who delivered Rh positive infants after 34 weeks in their first pregnancy in 1980-1 (group 1) and 2000 similar primigravidas from 1978-9 (group 2). Obstetric data were collected for 616 women in group 1 who had a subsequent pregnancy, 536 similar women in group 2, and 410 Rh positive but otherwise similar primigravidas who delivered in the same hospitals in 1978-81 (group C). INTERVENTIONS: Anti-D immunoglobulin 100 micrograms intramuscularly was given at 28 and 34 weeks to the mothers in their first pregnancy who delivered in 1980-1. END POINTS: Detection of anti-D antibody in the first or any subsequent pregnancy in groups 1 and 2. For all three groups having subsequent pregnancies gestation at delivery, birth weight, fetal survival at one month, pre-eclampsia defined as blood pressure greater than 140/90 on two occasions more than 12 hours apart, and proteinuria greater than 0.25 milligram. MEASUREMENTS AND MAIN RESULTS: Antenatal immunisation to Rh(D) occurred in six mothers in group 1 and 32 group 2. Most immunisations occurred in the first or second pregnancy. The rates of abortion, gestation at delivery, birth weight, and fetal survival were not significantly different among the three groups. The incidence of pre-eclampsia was lower in mothers given antenatal anti-D immunoglobulin, but the difference was not significant. CONCLUSIONS: Antenatal prophylaxis with anti-D immunoglobulin is effective, and the effect of giving it in the first pregnancy persists into at least the second pregnancy. It seems to be safe for the fetus in the index and subsequent pregnancies. PMID- 2547470 TI - Change in cough reflex after treatment with enalapril and ramipril. AB - OBJECTIVE: To find out whether enalapril or ramipril causes the sensitivity of the cough reflex to change or symptomatic cough to develop in patients with hypertension. DESIGN: Prospective, placebo controlled, double blind, randomised crossover study. SETTING: Academic units of clinical pharmacology and medicine. PATIENTS: 20 Patients (nine men and 11 women) who needed to take angiotensin converting enzyme inhibitors to control hypertension. INTERVENTIONS: All patients received enalapril 10 mg daily, ramipril 10 mg daily, or placebo daily for one week in random order, with a washout period of at least one week between treatments. For assessment of sensitivity of the cough reflex the patients inhaled various concentrations of capsaicin solution in random order. MAIN OUTCOME MEASURES: Measurement of the doses of capsaicin required to cause two or more and five or more coughs or the development of a symptomatic cough. RESULTS: Blood pressure, symptoms of cough, and the sensitivity of the cough reflex to inhaled capsaicin were recorded at the start of the study and before and at the end of each treatment period. Plasma urea and creatinine concentrations and angiotensin converting enzyme activity were measured at the start of the study and the end of each treatment period. Data were analysed by two way analysis of variance. Mean blood pressure was 159/97 mm Hg at the start of the study and 152/92, 143/88, and 147/86 mm Hg after treatment with placebo, enalapril, and ramipril respectively. Mean (SE) plasma angiotensin converting enzyme activity was 2.2 (0.2) mmol/l/h after treatment with placebo and fell significantly to 1.3 (0.1) mmol/l/h and to 0.4 (0.1) mmol/l/h after treatment with enalapril and ramipril respectively. No patient complained of cough while taking placebo but three women complained of cough when taking both enalapril and ramipril. The mean (95% confidence interval) lowest dose of capsaicin causing two or more coughs was 2.4 (1.5 to 4.0), 1.8 (1.12 to 2.82), and 2.2 (1.7 to 3.0) nmol after treatment with placebo, enalapril, and ramipril respectively; none of these changes were significant. The lowest dose of capsaicin causing five or more coughs was 18.9 (13.9 to 25.8), 14.4 (8.4 to 24.5), and 15.3 (10.8 to 21.2) nmol respectively; none of these changes were significant. The three patients who complained of cough had normal sensitivity to capsaicin after treatment with placebo but had a considerably increased sensitivity after treatment with enalapril and ramipril. CONCLUSIONS: Both enalapril and ramipril increase the sensitivity of the cough reflex appreciably in patients who complain of cough during treatment, but they do not change the se PMID- 2547471 TI - Nonfatal demyelinating encephalomyelitis induced by coronavirus (JHM strain) infection in mice. AB - C3H mice infected intravenously with the JHM strain of coronavirus showed high incidence of demyelination (44.8%) and low incidence of encephalitis-induced mortality (6.9%). High titers of virus were detectable in the brain and liver of mice only during the first 3 to 12 days of infection (10(3) and 10(4) PFU/g, respectively). Most of the animals recovered from the first phase of disease and some (11.1%) came down with paralysis 6 to 7 weeks after the infection, with no histological changes or virus detectable in their tissues. PMID- 2547472 TI - What confers specificity on glycine for its receptor site? AB - 1. The structural requirements for activation of the glycine receptor were studied in isolated ventromedial hypothalamic neurones of rats by use of a 'concentration-clamp' technique under single-electrode voltage-clamp conditions. 2. alpha-Amino acids (L-alpha-alanine, and D-alpha-alanine, and L-serine), and glycine-methylester, glycine-ethylester and beta-amino acids (beta-alanine and taurine) produced a transient inward Cl- current, which was similar to that induced by glycine. 3. The responses to individual alpha- and beta-amino acids were selectively antagonized by strychnine, but were not affected by bicuculline, picrotoxin or the taurine antagonist, TAG (6-aminomethyl-3-methyl-4H,1,2,4 benzothiadiazine-1,1-dioxide hydrochloride), suggesting that alpha- and beta amino acids activate the same glycine receptor. 4. beta-Amino acids were slightly more potent than the alpha-amino acids in causing cross-desensitization of the glycine response. 5. From the results of the structure-activity analysis of the optical isomers of alpha-alanine, serine and cysteine, a tentative structure of the glycine receptor is proposed. PMID- 2547473 TI - Inotropic and chronotropic effects of N6-substituted derivatives of cyclic AMP as assessed in guinea-pig isolated right atria and papillary muscle. AB - 1. The inotropic and chronotropic actions of N6-substituted adenosine 3':5' cyclic monophosphate (cyclic AMP) derivatives (N6-R cyclic AMPs) were studied in guinea-pig isolated right atrial preparations and in the papillary muscle preparations from guinea-pig right ventricle. 2. All the N6-R cyclic AMPs except N6-C14H29 produced positive inotropic effects in papillary muscle. The C5H11, C6H13, C7H15, C8H17 and C9H19 compounds were the most potent as inotropic agents, the potency being lower with compounds having longer or shorter N6-side chains than these. 3. In right atria N6-C2H5-C7H15 cyclic AMPs produced negative chronotropic effects. However, after treatment of the preparations with 8 phenyltheophylline the negative chronotropic effects were either much attenuated or abolished, indicating the involvement of adenosine receptors. 4. All the N6-R cyclic AMPs except N6-C14H29 were more potent activators of bovine myocardial protein kinase than cyclic AMP. The partition coefficients between octanol and an aqueous phase of N6-R cyclic AMPs became greater as the numbers of carbon atoms increased, and there appeared to be a relationship between partition coefficient and inotropic potency. It was concluded that membrane penetrativeness rather than potency as activators of protein kinase determined the potencies of N6-R cyclic AMPs as positive inotropic agents. 5. Derivatives such as N6-C7H15 cyclic AMP, which have positive inotropic activity without any marked negative chronotropic effect, may be useful as cardiotonic agents in heart failure. PMID- 2547475 TI - Mechanism of xanthine-induced relaxation of guinea-pig isolated trachealis muscle. AB - 1. Four 3-alkylxanthines (3-methylxanthine, 3-n-propylxanthine (enprofylline), 3 n-butylxanthine and 3-iso-butylxanthine) and four 1-methyl-3-alkylxanthines (1 methyl-3-methylxanthine (theophylline), 1-methyl-3-n-propylxanthine, 1-methyl-3-n butylxanthine and 1-methyl-3-iso-butylxanthine (IBMX], were compared in terms of cyclic AMP phosphodiesterase (PDE) inhibition and trachealis muscle relaxation. The relationship between xanthine structure and cyclic AMP PDE inhibition was also studied. 2. Xanthine induced relaxation of guinea-pig isolated trachealis muscle was measured against spontaneous tone. 3. The four 1-methyl-3 alkylxanthines were each significantly more potent than the corresponding 3 alkylxanthines in relaxing the isolated trachealis muscle. The 1-methyl-3 alkylxanthines were similarly more potent than the corresponding 3-alkyl derivatives in inhibiting low Km cyclic AMP PDE. There was a strong positive correlation between low Km cyclic AMP PDE inhibition and the tracheal smooth muscle relaxation evoked by the xanthine derivatives. 4. Since methylation of the 1-position of each 3-alkylxanthine increased the potency of the derivative in inhibiting low Km cyclic AMP PDE and in relaxing trachealis muscle and since a strong positive correlation was observed between the relaxant EC50 and the Ki value of each xanthine derivative, it is suggested that low Km cyclic AMP PDE inhibition by xanthines plays an important role in their tracheal relaxant effect. PMID- 2547474 TI - Arachidonic acid-induced mobilization of calcium in human neutrophils: evidence for a multicomponent mechanism of action. AB - 1. The mechanism(s) involved in the mobilization of calcium induced by arachidonic acid in human neutrophils was investigated. 2. The addition of arachidonic acid to a suspension of human neutrophils led to a time- and concentration-dependent mobilization of calcium which was the result of two separate and experimentally differentiable processes. The latter consisted of a rapid and transient phase followed by a slower and more sustained response. 3. The initial phase of calcium mobilization elicited by arachidonic acid was decreased in the presence of EGTA, inhibited by pertussis toxin as well as by nordihydroguaiaretic acid (NDGA), and diminished following a pre-incubation with leukotriene B4, but not platelet-activating factor. 4. The characteristics of the first phase of the mobilization of calcium were consistent with an interaction of the fatty acid with the leukotriene B4 receptors, either directly or indirectly following the synthesis of leukotriene B4, as well as with a release of internal calcium. 5. The second, slower and more sustained phase of calcium mobilization was more apparent at high concentrations (greater than or equal to 8-16 microM) of arachidonic acid, and was relatively insensitive to pertussis toxin, EGTA or NDGA. 6. The characteristics of the 'slow' phase of calcium mobilization by arachidonic acid are consistent with its being associated primarily with a release of calcium from internal storage pools. 7. The data presented indicate that the mechanism of mobilization of calcium by arachidonic acid in human neutrophils is complex and involves specific activation pathways employed, in part at least, by other neutrophil agonists. These findings may have relevance to various inflammatory situations in which the elevated levels of extracellular arachidonic acid known to be present could modulate the functional responsiveness of the neutrophils to other stimuli. PMID- 2547476 TI - The role of GABAA receptor function in peristaltic activity of the guinea-pig ileum: a comparative study with bicuculline, SR 95531 and picrotoxinin. AB - 1. The peristaltic activity of the guinea-pig ileum was studied in the absence and in the presence of the blockade of GABAA receptors. 2. Bicuculline (1-30 microM), improved at the highest concentrations the efficiency of peristalsis by enhancing the frequency of propulsive contractions and the amount of fluid ejected per unit of time. 3. Neither SR 95531 (0.3-10 microM), a novel GABAA receptor antagonist, which competitively antagonized 3-aminopropane sulphonic acid induced contractions in myenteric plexus-longitudinal muscle preparations (pA2 value: 6.47), nor picrotoxinin (1-30 microM) modified peristaltic parameters or influenced the potentiating effect of bicuculline on peristaltic activity. 4. In myenteric plexus-longitudinal muscle preparations, bicuculline (1-30 microM) enhanced the amplitude of electrically-induced cholinergic contractions without modifying submaximal contractions to applied acetylcholine. SR 95531 and picrotoxinin had no effect on twitch amplitude. In the presence of each of these compounds, bicuculline retained its potentiating effect. 5. The results obtained with SR 95531 and picrotoxinin question the view that GABAA receptors may exert a critical role in intestinal propulsion by modulating the activity of nerve pathways subserving peristalsis. Bicuculline potentiates the peristaltic activity of the ileum probably via a facilitatory effect on enteric cholinergic transmission that is independent of GABAA receptor blockade. PMID- 2547477 TI - Evidence for prazosin-resistant, rauwolscine-sensitive alpha-adrenoceptors mediating contractions in the isolated vascular bed of the rat tail. AB - 1. The postjunctional alpha-adrenoceptors mediating contractions in the isolated vascular bed of the perfused rat tail have been investigated, in the presence and absence of an increase in perfusion pressure by arginine vasopressin (AVP). 2. In the absence of AVP, bolus doses of noradrenaline (NA) and phenylephrine produced pressor responses of similar time course, while UK-14,304 was practically inactive. Responses to noradrenaline were inhibited more by 0.05 microM prazosin than by 1 microM rauwolscine, suggesting the presence of alpha1-adrenoceptors. 3. Following a sustained elevation in perfusion pressure by AVP, both UK-14,304 and NA (the latter in the presence of 0.05 microM prazosin to inhibit alpha 1 adrenoceptors) elicited dose-dependent pressor responses. The maximum response to UK-14,304 under these conditions was approximately 30% of the maximum response to NA in the absence of prazosin and AVP. Responses to phenylephrine were not affected by the AVP-induced increase in vascular tone. 4. In the presence of AVP, pressor responses to UK-14,304 were resistant to 0.05 microM prazosin and susceptible to antagonism by 1 microM rauwolscine (-log Kb 7.65 +/- 0.15). Similarly, responses to NA in the presence of 0.05 microM prazosin and AVP were inhibited by 1 microM rauwolscine. This represents the first demonstration of prazosin-resistant, rauwolscine-sensitive alpha 2-adrenoceptor-mediated responses in the vasculature of the rat tail. 5. These results suggest that in isolated vascular preparations, functional populations of postjunctional alpha 2 adrenoceptors may be 'uncovered' by the presence of AVP. PMID- 2547478 TI - Inhibition of bradykinin-induced bronchoconstriction in the guinea-pig by a synthetic B2 receptor antagonist. AB - 1. Intravenous bradykinin (Bk) elicited bronchoconstriction in the anaesthetized ventilated guinea-pig which was not mimicked by the B1 receptor agonist, des-Arg9 Bk. 2. Bradykinin-induced bronchoconstriction was inhibited by the B2 receptor antagonist B4881, but not by the B1 receptor antagonist des-Arg9-Leu8-Bk. The effect of B4881 was short-lived. 3. The B2 receptor antagonist as B4881 was selective for bradykinin as B4881 did not significantly inhibit bronchoconstriction induced by i.v. bombesin, platelet activating factor, acetylcholine, histamine or vagal stimulation. 4. These results suggest that bradykinin-induced bronchoconstriction in the guinea-pig is via activation of a B2 receptor population and that B4881 is a selective B2 antagonist that may be useful for investigating the involvement of bradykinin in the lung. PMID- 2547480 TI - Impairment of endothelium-dependent relaxation and changes in levels of cyclic GMP in aorta from streptozotocin-induced diabetic rats. AB - 1. Acetylcholine (ACh)-induced relaxation of aortic strips with endothelium and production of cyclic GMP between streptozotocin-induced diabetic and age-matched control rats were compared. 2. The concentration-response curve for ACh-induced relaxation was shifted to the right in diabetic rats. IC50 values for ACh were 4.57 +/- 0.67 x 10(-8) M and 1.00 +/- 0.87 x 10(-7) M in aortic strips from age matched control and diabetic rats, respectively (n = 6, P less than 0.05). 3. Relaxations produced by atrial natriuretic peptide (ANP) in diabetic aortae were similar to those in age-matched vessels. 4. Relaxations produced by sodium nitroprusside (SNP) in diabetic aortae were similar to those in age-matched vessels. 5. Basal levels of cyclic GMP and ACh-induced production of cyclic GMP were significantly decreased in diabetic rats. 6. These results suggest that functional changes in endothelium but not in guanylate cyclase activity in the aorta may occur in diabetes, and thus, spontaneous and ACh-induced formation of cyclic GMP may be decreased. This decrease in production of cyclic GMP may be responsible for the decreased response of the aorta to the relaxant effect of ACh. PMID- 2547479 TI - Inhibition of histamine-stimulated inositol phospholipid hydrolysis by agents which increase cyclic AMP levels in bovine tracheal smooth muscle. AB - 1. The effect on histamine-stimulated [3H]-inositol phosphate accumulation of a range of agents which increase the accumulation, or mimic the actions, of cyclic AMP has been investigated in bovine tracheal smooth muscle. 2. Salbutamol (1 microM), forskolin (1 microM) and vasoactive intestinal peptide (VIP, 1 microM) inhibited the inositol phosphate response to 0.1 mM histamine and increased the accumulation of [3H]-cyclic AMP in [3H]-adenine-labelled slices of bovine tracheal smooth muscle. The effect on inositol phospholipid hydrolysis was mimicked by the membrane permeant analogues of cyclic AMP, dibutrylcyclic AMP (1 mM) and 8-bromo-cyclic AMP (1 mM). 3. In contrast to salbutamol, which was equally effective at producing the two effects, forskolin produced large increases in [3H]-cyclic AMP accumulation (EC50 = 1.2 microM) at much higher concentrations than those required for inhibition of histamine-stimulated [3H] inositol phosphate accumulation (EC50 = 0.09 microM). However, significant increases in [3H]-cyclic AMP accumulation, of similar magnitude to those obtained with salbutamol and VIP, were observed over the concentration range appropriate for inhibition of the inositol phosphate response to histamine. 4. In the presence of histamine (0.1 mM), isobutylmethylxanthine (IBMX, 1 mM) and rolipram (0.1 mM) both significantly (P less than 0.05) elevated tissue [3H]-cyclic AMP levels. IBMX, rolipram and (to a lesser extent) SKF 94120 significantly (P less than 0.05) reduced histamine-stimulated [3H]-inositol phosphate accumulation by 81%, 68% and 20%, respectively. M&B 22948 was without a significant effect on either [3H]-cyclic AMP or histamine-induced [3H]-inositol phosphate accumulation. 5. Both rolipram and forskolin reduced the increase in incorporation of [3H] inositol into membrane phospholipids which followed stimulation with histamine. However, a significant inhibition of [3H]-inositol phosphate accumulation could be demonstrated under conditions in which there was no change in the level of [3H]-inositol incorporation. PMID- 2547481 TI - Differential control and calcium-dependence of production of endothelium-derived relaxing factor and prostacyclin by pig aortic endothelial cells. AB - 1. Production of endothelium-derived relaxing factor (EDRF) by primary cultures of pig aortic endothelial cells was assessed indirectly by measuring endothelial cyclic GMP content, and prostacyclin production was measured by radioimmunoassay of 6-keto prostaglandin F1 alpha (6-keto PGF1 alpha). 2. The resting level of cyclic GMP fell significantly following removal of extracellular calcium (1 mM EGTA present), but elevations of cyclic GMP content induced by sodium azide (10 microM) or atriopeptin II (10 nM) were similar in the absence and presence of extracellular calcium. 3. Haemoglobin (10 microM) reduced the resting level of cyclic GMP in the presence, but not the absence of extracellular calcium. M&B 22,948 (100 microM), superoxide dismutase (30 u ml-1), bradykinin (0.1 microM), ATP (10 microM) and ionophore A23187 (0.1 microM) each induced an increase in endothelial cyclic GMP content that was reduced in the absence of extracellular calcium. 4. In cascade bioassay experiments using endothelial cells on microcarrier beads and perfused in columns, continuous infusion of bradykinin (0.1 microM) induced release of EDRF, assayed on rabbit aortic rings, that was maximal after 2 min and still detectable up to about 16 min. 5. In the presence of extracellular calcium, the time course of bradykinin (0.1 microM)-stimulated production of EDRF, assessed as endothelial cyclic GMP content was maximal within 1 min, declined thereafter, but was still significant after 30 min. Production of 6-keto PGF1 alpha, measured simultaneously rose rapidly but was complete within 3 min. 6. In the absence of extracellular calcium the resting endothelial content of cyclic GMP fell, but resting production of 6-keto PGF1 alpha was unaffected. 7. In the presence of TMB-8 (100 microM) resting endothelial content of cyclic GMP rose slightly, but production of 6-keto PGFg fell. The bradykinin (0.1 microM)-stimulated increase in cyclic GMP content was augmented, but the stimulation of 6-keto PGF1I production was blocked. Results from cascade bioassay experiments confirmed that TMB-8 (100 microM) did not inhibit bradykinin-induced production of EDRF. 8. The data suggest that resting production of EDRF but not prostacyclin is dependent upon the presence of extracellular calcium. Bradykinin stimulated production of EDRF is sustained and requires the presence of extracellular calcium, but stimulated production of prostacyclin is transient and may result from discharge of an intracellular pool of calcium. 9. The vascular endothelial cell appears therefore to control differentially production of EDRF and prostacyclin. PMID- 2547482 TI - Alpha 1-adrenoceptor function and autoradiographic distribution in human asthmatic lung. AB - 1. The autoradiographic distribution of alpha 1-adrenoceptors was investigated in non-diseased and asthmatic human lung by use of [3H]-prazosin (H-PZ). To validate binding and autoradiographic methods, H-PZ binding was also measured in rat heart. 2. Significant levels of specific H-PZ binding were detected in sections of rat heart. This binding was associated with a single class of non-interacting sites of high affinity (dissociation constant, Kd = 1.17 +/- 0.26 nM). The maximum binding capacity (Bmax) was 59.5 +/- 4.5 fmol mg-1 protein. 3. In sharp contrast, very low levels of specific H-PZ binding were found in both human nondiseased and asthmatic bronchus, although a high level of binding of [125I] iodocyanopindolol (I-CYP, 50 pM) to beta-adrenoceptors was detected in these airways. Furthermore, very low levels of autoradiographic grains representing specific H-PZ binding were found in all airway structures in human non-diseased or asthmatic lung parenchyma. 4. Consistent with these data, the alpha adrenoceptor agonist phenylephrine failed to induce significant increases in tone in bronchi isolated from either non-diseased or asthmatic human lung. Results indicate that asthma does not involve significant increases in airway alpha 1 adrenoceptor function. PMID- 2547483 TI - Developmental disappearance of excitatory alpha 1-adrenoceptor function in the oesophagus of chick embryo. AB - 1. Developmental change in the response to noradrenaline (NA) and 5 hydroxytryptamine (5-HT) was investigated in the chick oesophagus between 9 and 21 days of incubation and 4 days after hatching. 2. NA (5 microM) produced a significant contraction in the oesophagus at 9 days of incubation. The NA-induced contraction progressively decreased with development and changed to an inhibition of spontaneous contraction or a small relaxation by 17 days of incubation. 3. The NA-induced contractile response was inhibited by phentolamine (2.7 microM) and prazosin (0.55 microM). Phenylephrine (5 microM) but not clonidine (5-50 microM), also induced a contraction at early stages. The relaxation response to NA was sensitive to the beta-receptor blocker, carteolol (3.4 microM). 4. Pretreatment with carteolol unmasked the contractile responses to NA in preparations at 17-19 days of incubation. However, even in the presence of carteolol, the contraction produced by NA decreased and disappeared by the time of hatching. This change in response to NA is accompanied by a decline in the pD2 value. The response to phenylephrine (5 microM) followed the same pattern as that to NA. 5. The maximum binding sites of [3H]-dihydroergocryptine to the crude membrane preparation from oesophagus changed little at 13, 17 and 21 days of incubation. 6. Isoprenaline (Iso, 0.01-20 microM) caused a carteolol-sensitive relaxation in the carbachol contracted oesophagus after 13 days of incubation. The sensitivity (pD2 value) to Iso decreased slightly up to 17 days of incubation. 7. 5-HT (10 microM) caused a contraction in the oesophagus after 13 days of incubation and the amplitudem of the response increased up to 17 days of incubation. The response to 5-HT was abolished by methysergide (1 microM) but not by tetrodotoxin (0.78 microM) or atropine (1 microM) at every stage tested. 8. These results suggest that the response to NA changed from an alpha-adrenoceptor-mediated contraction to a beta receptor-mediated relaxation during the embryonic period, resulting partly from the decline and disappearance of excitatory alpha-receptor function in the chick oesophagus. PMID- 2547485 TI - Kappa-opioid-induced changes in renal water and electrolyte management and endocrine secretion. AB - 1. Subcutaneous injection of the kappa-opioid agonist U50,488 into conscious, saline-loaded rats was associated with a diuresis, antinatriuresis and antikaliuresis which lasted for up to 3 h. Plasma renin activity and corticosterone levels were elevated but plasma vasopressin (AVP) and aldosterone levels were unaltered in similarly treated rats. 2. U50,488 administration to adrenal demeddulated rats was not associated with a diuresis but produced an antinatriuresis, though sodium excretion rates were higher in demedullated than in sham-operated animals. Plasma AVP and corticosterone levels were not affected by demeddulation or subsequent U50,488 treatment. Sham-operated, U50-488-treated rats showed the expected increase in plasma corticosterone levels. 3. U50,488 administration resulted in an antidiuresis and an antinatriuresis in AVP deficient Brattleboro DI rats. 4. When coupled with fasting stress U50,488 administration resulted in similar but attenuated renal responses compared with those observed in unfasted rats. Basal plasma corticosterone levels were elevated in fasted animals and were further increased by U50,488. 5. Both water and electrolyte handling by the kidney are altered by U50,488. The diuretic effects of U50,488 were reversed by adrenal demedullation and in the absence of endogenous AVP, but the antinatriuretic actions were not altered, suggesting that the effects upon renal water and electrolyte excretion may be mediated by separate mechanisms. PMID- 2547486 TI - The effects of drugs interacting with opioid receptors on the early ventricular arrhythmias arising from myocardial ischaemia. AB - 1. The effects of a range of opioid receptor agonists and antagonists with differing opioid receptor selectivities on ischaemia-induced arrhythmias in anaesthetised rats was investigated. 2. Naloxone was antiarrhythmic only at doses expected to antagonise kappa- and delta-receptors in addition to mu-receptors. 3. The opioid receptor antagonist Mr 2266, which is twice as potent at kappa receptors as at mu-receptors dose-dependently reduced the incidence and severity of the arrhythmias resulting from coronary artery occlusion. 4. The opioid receptor antagonist M 8008 (1 mg kg-1), which is twice as potent at delta receptors as at mu-receptors but has very little affinity for the kappa-receptor, did not exhibit any beneficial antiarrhythmic properties. 5. MrZ 2593, a quarternary complex of naloxone which does not readily cross the blood brain barrier, was antiarrhythmic which implies that the antiarrhythmic actions of opioid receptor antagonists may be mediated via peripheral opioid receptors. 6. The agonists, diamorphine, [Leu] enkephalin and U-50,488H exhibited no significant arrhythmogenic effects under the present experimental conditions. 7. It is tentatively suggested that blockade of peripheral kappa-receptors during acute myocardial ischaemia may result in an antiarrhythmic effect. PMID- 2547484 TI - Central nervous system kinin receptors and the hypertensive response mediated by bradykinin. AB - 1. Bradykinin (Bk) administered intracerebroventricularly to the rat causes an increase in arterial pressure. 2. Analogues of Bk with agonist and antagonist activity were injected, over a wide dose-range, into the posterior region of the fourth ventricle of unanaesthetized rats implanted with permanent ventricular canullae, and blood pressure was measured directly from the abdominal aorta. 3. The analogues Ile-Ser-Bk (T-kinin) and Lys-Lys-Bk, which interact with both B1 and B2 Bk receptors, produced pressor effects similar to those of Bk, although of greater duration, whereas des-Arg9-Bk, a B1-receptor agonist, had no effect. 4. The B1-antagonist des-Arg9-[Leu8]-Bk did not alter the Bk pressor response, but D Arg-[Hyp3, Thi5,8,D-Phe7]-Bk, which interacts both with B1- and B2-receptors blocked the responses to Bk, T-kinin and Lys-Lys-Bk and caused parallel shifts to the right of the Bk dose-response curves. Neither antagonist, by itself, had any effect on blood pressure. 5. It is concluded that the central pressor response to Bk is mediated by receptors of the B2 subtype. PMID- 2547487 TI - Postjunctional alpha 2-adrenoceptors mediate vasoconstriction in human subcutaneous resistance vessels. AB - 1. In vitro studies have been performed on human medium-sized muscular arteries (internal diameter 1-4 mm) in a classical organ bath and with human subcutaneous resistance arteries (internal diameter 103-626 microns) in a microvascular myograph. 2. Although the medium-sized muscular arteries showed no response to either of the alpha 2-agonists B-HT 933 or UK 14304 in concentrations up to 10 microM, the subcutaneous resistance arteries from all regions examined showed well-pronounced and concentration-dependent responses to B-HT 933, the pD2 (-log EC50) being 5.11 +/- 0.09. 3. In the resistance arteries the alpha 2-antagonist yohimbine caused a parallel shift to the right of the B-HT 933 concentration response curve; the yohimbine pA2 for the B-HT 933 receptor was 7.86 +/- 0.12. 4. There was an inverse relationship between the maximum response to B-HT 933 and the calibre of the resistance vessels. 5. These results indicate the presence of a postjunctional alpha 2-adrenoceptor in human subcutaneous resistance arteries and not in medium sized muscular arteries. PMID- 2547488 TI - Effect of endothelium on basal and on stimulated accumulation and efflux of cyclic GMP in rat isolated aorta. AB - 1. The aim of this study was to examine the possible role of the release of guanosine 3':5'-cyclic monophosphate (cyclic GMP) into the extracellular space in the regulation of rat aortic cyclic GMP content. 2. Rat aortic segments incubated in physiological solution released cyclic GMP into the medium in a time-dependent manner. This release was greatly enhanced when intact instead of tissues without endothelium were used. After 120 min of observation, a maximal 33 fold difference in extracellular cyclic GMP content was detected. 3. Treatment of rat aortic preparations with either a Ca2+-free solution or methylene blue, both conditions known to inhibit endothelium-derived relaxing factor (EDRF)-mediated responses, markedly reduced the extracellular accumulation of cyclic GMP from tissues with but not without endothelium. 4. Endothelium-dependent vasodilators such as acetylcholine (10 microM) and carbachol (10 microM) greatly increased tissue cyclic GMP content, in a time-dependent manner in rat aortic preparations with endothelium, but only slightly in tissues without. Maximal increases in intact tissues were obtained after about 1 min of agonist contact and amounted to about 35 and 15 fold respectively, thereafter tissue cyclic GMP content rapidly declined. Histamine (10 microM) elicited only minor effects on tissue cyclic GMP content of both intact preparations and those without endothelium. 5. Acetylcholine (10 microM), carbachol (10 microM) and histamine (10 microM) stimulated a time-dependent release of the cyclic nucleotide into the incubation medium from tissues with endothelium. After 120 min of observation, extracellular accumulation of cyclic GMP from intact tissues was increased by about 2.6, 6.6 and 1.7 fold respectively. Carbachol and histamine induced only minor effects on release from tissues without endothelium. 6. Sodium nitroprusside (0.3 and 10 microM), a direct activator of soluble guanylate cyclase, induced a concentration dependent accumulation of cyclic GMP in tissues with and without endothelium that was associated with a concentration-dependent accumulation of cyclic GMP in the extracellular space. Peak tissue cyclic GMP content reached similar levels in preparations with and without endothelium, while extracellular cyclic GMP levels were about two times greater when experiments were performed with intact compared to endothelium-denuded tissues. 7. Atriopeptin II, an activator of particulate guanylate cyclase, increased tissue cyclic GMP content by about 8 and 18 fold respectively in tissues with and without endothelium. As was the case with sodium nitroprusside, atriopeptin II-stimulated release was markedly enhanced from intact tissues compared with those without endothelium. After 120 min of observation, there was a 16 fold difference in the amount of extracellular cyclic GMP. PMID- 2547489 TI - Potassium channel blockers differentially affect carbachol and (-)-N6 phenylisopropyladenosine on guinea-pig atria. AB - 1. The effect of three different potassium channel blockers (tetraethylammonium, TEA; 4-aminopyridine, 4-AP; and apamin) and of variations in the concentration of K+ and Ca2+ in the medium, have been studied on the responses of guinea-pig isolated atria to (-)-N6-phenylisopropyladenosine (R-PIA), a stable adenosine A1 receptor agonist, and to carbachol, a muscarinic agonist. R-PIA and carbachol showed the same negative inotropic effects over a similar range of concentrations (3-300 microM), both in spontaneously beating and in electrically driven atria. 2. TEA (0.1 to 20 mM) and 4-AP (0.3 to 3 mM), both antagonized the negative inotropic and chronotropic effects of carbachol in a concentration-dependent manner. In contrast, these compounds failed to inhibit the effects induced by R PIA. Apamin, a specific blocker of a low conductance Ca2+-activated K+ channel, was ineffective in accordance with the absence of these channels in atrial tissue. 3. TEA (0.1 to 20mM) inhibited the negative inotropic effect of carbachol, but not that of R-PIA, in atria paced and depolarized by a high K+ medium (22 mM). In this preparation Na+ current is abolished and the contraction induced by noradrenaline and electrical stimulation is solely dependent on Ca2+ influx currents. 4. Stepwise addition of Ca2+ to a calcium-depleted perfusing medium of electrically driven atria, induced a positive inotropic effect which was inhibited by R-PIA. In contrast, carbachol had no effect. 5. In agreement with our previous study, the data suggest that R-PIA acts on isolated atria by inhibiting Ca2+ influx through L-channels. PMID- 2547490 TI - Effects of adenosine on polymorphonuclear leucocyte function, cyclic 3': 5' adenosine monophosphate, and intracellular calcium. AB - 1. Inhibition of human polymorphonuclear leucocyte (PMN) function by adenosine was studied with respect to effects of adenosine on intracellular cyclic AMP and calcium during the PMN respiratory burst. 2. The adenosine analogue 5'-N ethylcarboxamide-adenosine (NECA) and L-N6-phenyl-isopropyl-adenosine (L-PIA) inhibited PMN oxygen metabolite generation with relative potencies (NECA greater than adenosine greater than L-PIA) characteristic of an A2 receptor. 3. The respiratory burst was inhibited by adenosine when PMN were activated by calcium ionophore or chemotactic peptide but not when cells where activated by oleoyl acetyl-glycerol (OAG). 4. Adenosine increased intracellular cyclic AMP during the PMN respiratory burst regardless of whether cells were stimulated by ionophore, chemotactic peptide or OAG. 5. To determine whether the differences in cell inhibition by adenosine were related to differences in intracellular calcium mobilization by each activating agent, calcium was evaluated with the fluorescent probe, indo-1. Adenosine suppressed the increase in intracellular calcium following PMN activation by calcium ionophore or chemotactic peptide. In contrast, calcium did not increase in PMN activated by OAG and adenosine did not affect intracellular calcium changes following this stimulus. 6. These results demonstrate that physiological concentrations of adenosine inhibit the PMN respiratory burst in association with an increase in intracellular cyclic AMP and reduction of intracellular calcium. PMID- 2547491 TI - Block of sodium channels by psychotropic drugs in single guinea-pig cardiac myocytes. AB - 1. Effects of imipramine and haloperidol on voltage-gated sodium channels were investigated in guinea-pig isolated ventricular myocytes by the whole-cell patch clamp technique. Some additional experiments were also performed with chlorpromazine for the purpose of comparison. 2. All test drugs in micromolar concentrations suppressed the amplitude of peak sodium current associated with step depolarization from a holding potential of -140 mV in a reversible manner. The order of potency was chlorpromazine greater than imipramine greater than haloperidol. 3. Dose-response curves obtained with a holding potential of -140 mV were best fitted by 2:1 stoichiometry in all three drugs and were shifted in the direction of lower concentrations when a holding potential of -90 mV was used. 4. The drug-induced block was not associated with any change in the time courses of sodium current activation and inactivation. 5. Steady-state sodium channel inactivation curve was shifted in the direction of more negative potentials by the drugs. 6. All three drugs also produced marked use-dependent block as demonstrated by a cumulative increase in the block during a train of depolarizing pulses. 7. The use dependence was due to a higher affinity of the drugs for the inactivated state of sodium channels than the resting state and to a very slow repriming of the drug-bound sodium channels from inactivation. 8. The steady state and use-dependent block of voltage-gated sodium channels by psychotropic drugs may contribute to their cardiotoxic and perhaps antiarrhythmic effect. PMID- 2547492 TI - Use of geographutoxin II (mu-conotoxin) for the study of neuromuscular transmission in mouse. AB - 1. Endplate potentials (e.p.ps) were investigated in the presence of geographutoxin II (GTXII) in the mouse phrenic nerve diaphragm preparation. This toxin preferentially blocks muscle Na+ channels which allows the study of e.p.ps in the absence of nicotinic receptor antagonists or substances to depress acetylcholine release. 2. GTXII abolished muscle action potentials and antagonized the depolarization of the muscle membrane produced by the crotamine induced opening of Na+ channels. 3. E.p.ps as large as 19-25 mV were observed after 2-4 micrograms ml-1 GTXII. These concentrations of GTXII did not cause discernible changes of resting membrane potential and frequency and amplitude of miniature e.p.ps. 4. Lower concentrations (1-2 micrograms ml-1) of GTXII caused incomplete blockade of the muscle Na+ channel resulting in exaggerated 'e.p.ps', while higher concentrations of GTXII (8 micrograms ml-1) abolished e.p.ps by a prejunctional effect. 5. Trains of e.p.ps on repetitive stimulation after GTXII neither ran down, as in tubocurarine-treated preparations, nor facilitated, as in low Ca2+ and/or high Mg2+-treated preparations, and were indistinguishable from those of untreated cut muscle preparation. 6. In cut muscle preparations, GTXII did not affect the rise and decay times, amplitude or rundown of e.p.ps. 7. It is concluded that GTXII is a useful agent for studying neuromuscular transmission. This method provides e.p.ps which are neither attenuated nor modified because manipulations that alter transmitter release and postjunctional receptor responses are avoided. PMID- 2547493 TI - The dihydropyridine niguldipine modulates calcium and potassium currents in vascular smooth muscle cells. AB - 1. Vascular smooth muscle cells were isolated from the portal vein and from pial vessels of the cow. They were voltage-clamped with a single patch electrode technique (whole cell recording) in order to analyse the effects of niguldipine on ionic membrane currents. Due to adsorption of niguldipine to plastic and glass, the effective concentrations are lower than the nominal concentrations by a factor of about 3. 2. Niguldipine reduced Ca-currents (ICa of the L-type, voltage operated) at nominal concentrations greater than 0.1 microM up to a complete block at 1 microM (50% block at 0.4 microM). Nominal concentrations between 50 and 200 nM facilitated ICa ('Ca-agonistic effect'). The Ca-agonistic effects of niguldipine showed modest use- but strong voltage-dependence. 3. Niguldipine increased the outward currents at nominal concentrations greater than 10 nM. The extra outward currents reversed at -85 mV, the result suggesting that niguldipine had increased potassium currents, IK. Maximal facilitation of IK by niguldipine was about 400% and was obtained at 1 microM, half-maximal facilitation was obtained with a nominal concentration of 20 nM. 4. Both reduction of ICa and facilitation of IK may contribute to vasodilatation by niguldipine. Due to its greater sensitivity, the effects on IK may dominate. PMID- 2547495 TI - Hepatic tumour resection with profound hypothermia and circulatory arrest. PMID- 2547496 TI - Management of screen detected ductal carcinoma in situ of the female breast. AB - Thirty eight ductal carcinomas in situ of the female breast detected during the first 7 years of screening by the Guildford Breast Screening Unit have been treated by one surgeon. Twenty-eight cases were treated conservatively and ten by mastectomy. In the group treated conservatively there have been five local recurrences: four as ductal carcinoma in situ and one as node negative microinvasive carcinoma. There were no clinical or pathological features that predicted local recurrence, which was detected only by follow-up mammography. Based on these early results, an initially conservative approach to screen detected ductal carcinoma in situ is advocated. PMID- 2547494 TI - Actions of guanine nucleotides and cyclic nucleotides on calcium stores in single patch-clamped smooth muscle cells from rabbit portal vein. AB - 1. Single smooth muscle cells were obtained from the rabbit portal vein by enzymic digestion and membrane currents under voltage clamp measured by whole cell patch clamp technique. 2. When held at depolarized potentials, spontaneous outward currents (STOCs) were discharged; it is likely that these represent the cyclical storage and release within the cell of calcium in relation to Ca activated K-channels. 3. Application of lower concentrations of carbachol (10( 5)M) or caffeine (10(-3)M) accelerated STOC discharge. Higher concentrations of caffeine (10(-2)M) or carbachol (10(-4)M), or noradrenaline (10(-5)M), produced an outward current of 1-5 nA which disappeared within 5-15s and which was considered to result from the discharge of calcium stores; STOC discharge was abolished for a period. 4. Ryanodine (10(-5)-10(-4)M) or a non-hydrolysable GTP analogue, GTP gamma S (10(-5)-10(-3)M) introduced into the cell abolished STOC discharge within 2-5 min. STOCs were large in cells filled with GDP beta S (10( 3)M) and the action of GTP gamma S introduced at various concentrations was antagonized. 5. GTP gamma S (10(-4)-10(-3)M) in the cell reduced or abolished outward current to caffeine (10(-2)M) noradrenaline (10(-5)M) or carbachol (10( 4)M); the effect on caffeine outward current was antagonized by GDP beta S (10( 3)M) introduced into the cell. GDP beta S reduced noradrenaline outward current but not caffeine outward current implying the existence of a G-protein step in noradrenaline-evoked Ca-store release, possibly regulating phospholipase C enzyme activity and D-myo inositol 1,4,5 trisphosphate formation. 6. If cyclic AMP (10( 3)M) or cyclic GMP (10(-3)M) was introduced into the cell, or 8-bromo cyclic AMP (0.5 x 10(-3)M) or 8-bromo cyclic GMP (0.5 x 10(-3)M) applied to the cell in the bathing solution, STOC discharge was only slightly affected. However, the outward current to caffeine applied after noradrenaline was much enhanced. 7. The results could be explained if cyclic GMP and cyclic AMP enhance calcium storage whereas GTP gamma S depletes calcium stores, an action antagonized by GDP beta S. PMID- 2547497 TI - Capsular penetration and perforation in pleomorphic adenoma of the parotid salivary gland. AB - Both penetration and perforation of the capsule were seen on the superficial aspect of pleomorphic adenomas. Only penetration was noted on the deep aspect, where the tumour was adjacent to the facial nerve. In terms of local recurrence this may explain why it appears to be safe to separate the deep aspect of the tumour from the facial nerve without a wide margin of normal parotid tissue. PMID- 2547498 TI - Hippocampal kindling enhances excitatory amino acid receptor-mediated polyphosphoinositide hydrolysis in the hippocampus and amygdala/pyriform cortex. AB - We recently demonstrated that a long-lasting increase in ibotenate-stimulated polyphosphoinositide (PPI) hydrolysis in the amygdala/pyriform cortex (AM/PC) is associated with seizure susceptibility of amygdala (AM)-kindled rats. The present study examined (1) whether ibotenate-stimulated PPI hydrolysis would be lastingly enhanced in the hippocampus (HIPP) and AM/PC of the HIPP-kindled rats and (2) whether similar changes would be found in the early stage of HIPP kindling. Although ibotenate-stimulated accumulation of [3H]inositol 1-phosphate ([ 3H]IP1) increased significantly in the HIPP 24 h, 5 days, and 15 days after the last seizure of fully developed HIPP-kindled rats, no statistically significant increase was found in the HIPP 30 days after the last seizure. In the AM/PC, 10( 3) M ibotenate-stimulated [3H]IP1 accumulation significantly increased by 91%, 91%, 86% and 90%, 24 h, 5 days, 15 days and 30 days after the last seizure, respectively. There was no significant increase in ibotenate-stimulated [3H]IP1 accumulation 7 days after the last stimulation in the HIPP and AM/PC of rats which had undergone electrical stimulation only 5 times in the HIPP. These results indicate that (1) PPI hydrolysis coupled to excitatory amino acid receptors increases long-lastingly in the AM/PC regardless of the primary kindled site, and (2) these changes do not occur in the early stage of HIPP-kindling. PMID- 2547499 TI - Rapid decrease of high affinity ouabain binding sites in hippocampal CA1 region following short-term global cerebral ischemia in rat. AB - High affinity [3H]ouabain binding was examined in the hippocampal CA1 region and frontal cortex of rats subjected to 5 min complete cerebral ischemia in a clinical death model, and to subsequent resuscitation. A decrease of Bmax directly after ischemia and its further gradual decrease during 120 min of reperfusion were noted in the ischemia-vulnerable CA1 region, whereas no change of Bmax was observed in frontal cortex. The apparent Kd constant showed insignificant fluctuations in either of the two brain regions. Since ouabain binds with high affinity to the neuronal (alpha +)-form of Na+/K+-ATPase, the results indicate a rapid enzyme loss in CA1 neurons. The high affinity ouabain binding test proved to be a sensitive detector of premorphological changes in nerve cell membranes in ischemia. PMID- 2547500 TI - Liver cancer. PMID- 2547501 TI - Higher plasma Na+,K+-ATPase inhibitory activity in essential hypertensive patients. AB - The ability of plasma to inhibit 86 rubidium uptake in rat aorta and to displace [3H]-ouabain from hog brain Na+,K+-ATPase was used as a measure of plasma Na+,K+ ATPase inhibitory activity in seven normotensive and eight hypertensive subjects. Rat aortae rings were incubated in oxygenated plasma containing 86 rubidium (2 microCi/mL) for 30 mins at 37 degrees C and uptake measured and expressed as mumol/kg wet weight/min. Plasma was extracted with a mixture of chloroform and methanol (2:1) and the extract separated by silicic acid column followed by thin layer chromatography and fractions assayed for ouabain displacement using digoxin as a standard. Total ouabain displacement was calculated as the sum of all fractions. There was a strong correlation between the two methods for total plasma Na+,K+-ATPase inhibitory activity (r = 0.761, P less than 0.01). There was a significant positive correlation between plasma Na+,K+-ATPase inhibitory activity and blood pressure in all subjects. Na+,K+-ATPase inhibitory activity was significantly higher in plasma of hypertensives by both methods (P less than 0.001). The increased Na+,K+-ATPase inhibitory activity in plasma from hypertensives was due to the nonesterified fatty acid, long chain acylcarnitine and diphosphatidylglycerol fractions. PMID- 2547502 TI - Role of cGMP in relaxation of vascular and other smooth muscle. AB - The hypothesis that the relaxant action of many drugs on vascular and other smooth muscle is mediated by increases in intracellular cGMP, the "cGMP hypothesis," is gaining wide acceptance. While much information supporting this idea can be found in the literature, there is also a significant amount of information indicating that an elevation in the tissue content of cGMP is by itself insufficient to cause smooth muscle relaxation. The literature is reviewed with reference to the criteria that need to be fulfilled to consider cGMP as the second messenger mediating relaxation of smooth muscle by a drug; i.e., activation of guanylate cyclase, elevation of tissue content of cGMP, potentiation by phosphodiesterase inhibitors, antagonism by inhibitors of cGMP synthesis, and production of relaxation by cGMP analogues. For each criterion, key observations supporting the hypothesis are considered, followed by examples of important observations not consistent with the hypothesis. It is concluded that in some smooth muscles, for example, rat myometrium and vas deferens, cGMP is not a mediator of drug-induced relaxation. In other smooth muscles, including vascular smooth muscle, cGMP appears to play an important role in the relaxation process; but current evidence suggests that other factors are also important and that the cGMP hypothesis may need to be modified. PMID- 2547503 TI - The action and metabolism of peptide leukotrienes in the isolated bullfrog lung. AB - Leukotrienes (LTs) have been shown to cause contraction of mammalian airway smooth muscle. In this study, LTC4, LTD4, and LTE4 were tested on isolated strips of bullfrog lung. LTC4, LTD4, and LTE4 (10(-7) to 3 x 10(-10) M) stimulated lung contraction. LTC4 was the most potent, with LTD4 and LTE4 being of equal but lower potency. The cyclooxygenase inhibitors, indomethacin and ibuprofen, had no effect on the strength of leukotriene-induced contraction. In addition, the effects of three mammalian LTD4 receptor antagonists, L-649,923, L-648,051, and LY171883 were tested. All three antagonists failed to block LTC4-simulated contraction, but were effective blockers of LTD4. LTE4-stimulated contractions were significantly blunted by all three antagonists, but the extent of blockade was less than for LTD4. Significant bioconversion of [3H]LTC4 to LTD4 and LTE4 occurred in minced lung preparations but not in lung strips. Peptide leukotrienes caused contraction in amphibian lung, though the order of potency differs from mammals. Like mammals, frogs appear to have two classes of leukotriene receptors, one for LTC4 and one for LTD4-LTE4. These results support the hypothesis that leukotriene receptors have been highly conserved through evolution, despite the fact that the nature of tissue responsiveness to leukotrienes has changed over evolutionary time. PMID- 2547504 TI - Hypothalamic cyclic AMP after the injection of ovarian steroids into ovariectomized rats. AB - The effect of ovarian steroids on the concentration of adenosine 3',5'-cyclic monophosphate (cAMP) in the hypothalamus was studied in ovariectomized rats. Ovariectomized rats exhibited a lower cAMP concentration than intact rats. The administration of a single dose of estradiol benzoate (50 micrograms/kg body weight) resulted 3 days later in a rise of cAMP values, but levels did not reach those observed in estrous rats. Progesterone (2 mg/rat) injected 3 days after the priming dose of estradiol benzoate produced 4 h later no further changes in hypothalamic cAMP. The changes in hypothalamic cAMP concentration induced by estrogen treatment depend, at least in part, on noradrenergic inputs, since they were prevented by the injection of the norepinephrine synthesis inhibitor, diethyldithiocarbamate. In addition, administration of the beta-blocking agent, propranolol, to estradiol- and estradiol-progesterone-treated rats lowered the concentration of cAMP in the hypothalamus in a dose-dependent manner. In contrast, the administration of an alpha-blocking agent, phenoxybenzamine, had no effect at the tested concentration. The results of this study indicate that estrogen increases cAMP concentration in the hypothalamus by a noradrenergic mechanism involving beta-receptors. Moreover, the findings suggest that estrogen induces an increase in the number of beta-receptor sites, whereas progesterone increases the apparent propranolol sensitivity for these receptor sites. PMID- 2547506 TI - Histopathology and prognosis in childhood hepatoblastoma and hepatocarcinoma. AB - To determine the relationship between outcome and histologic type, the authors examined data from 168 cases of hepatoblastoma (HB) and 28 cases of hepatocarcinoma (HC) accrued from children over a 14-year period. After adjustment for stage of disease, there was no significant difference in median survival between HB and HC. Mitotic activity was associated with poor prognosis. Necrosis or vascular invasion did not influence prognosis. In 55 cases of completely resected HB, pure fetal histologic type (PFH) was associated with improved survival when compared with all other histologic patterns of HB (92% versus 57% 24 months' survival; P = 0.02). A prognostic effect of PFH was not demonstrable in incompletely resected HB, but the absence of mitoses and the presence of differentiated mesenchymal elements improved survival. The fibrolamellar pattern of HC demonstrated survival similar to that of the typical pattern of HC. The authors conclude that features consistent with differentiation in HB convey improved prognosis for survival. These observations may be important in designing future therapy for children with hepatic tumors. PMID- 2547505 TI - Concomitant 5-fluorouracil infusion and high-dose radiation for stage III non small cell lung cancer. AB - Thirty patients with Stage III non-small cell lung cancer were entered on a trial to evaluate the feasibility of combined radiation and concomitant 5-fluorouracil infusion. Patients had received prior debulking surgery (nine), induction chemotherapy (16), or no therapy (five). Radiation employed standard fractionation (180-200 rad/day) administered to a median cumulative dose of 5500 rad (range, 4500-6200 rad). 5-Fluorouracil was infused 24 hours per day throughout the period of radiation at a dose of 300 mg/m2/day for a median of 42 days (range, 28-56 days). Radiation complications included pneumonitis three of 30 (10%) and esophagitis (27%). Chemotherapy complications included stomatitis, two of 27 (7%), and hand-foot syndrome, three of 30 (10%). Treatment interruptions were necessary in six of 30 (20%) and four of 30 required parenteral nutrition. At a median follow-up of 12 months 26/30 (87%) maintained local control and eight had distant metastases (three of whom presented with Stage IV disease). 5-Fluorouracil delivered continuously throughout standard fractionation radiation to high cumulative doses is feasible and practical. Comparative clinical trials of the various combined radiation and chemotherapy schedules employed are in order. One additional clinical observation was the identification of six of 30 (20%) with brain metastases at presentation or after 12 months, all of whom had adenocarcinoma histologic subtype. PMID- 2547507 TI - An assessment of the mucous component in carcinoma of the colon and rectum. AB - It has been said that the prognosis of mucinous carcinoma of the colon and rectum is unfavorable. To determine the clinicopathologic significance of carcinomatous lesions with marked mucous production, the ratio of the area where there was significant mucous production (mucous component [MC]) to the area of the whole tumor on a tissue slice (area ratio) was determine by measurement with a digitizer in 281 patients with carcinoma of the colon and rectum. The MC was observed in the largest cross-section of the tumor in 85 patients (30.2%). Of these 85 patients the MC area ratio was less than 10% in 42. In the 43 patients with an area ratio of 10% or more, no particular relationship was observed between the area ratio and the frequency of cases. If no MC was observed in the largest cross section of the tumor, the MC ratio in other regions of the tumor was almost 0. In the patients in whom the MC area ratio in the largest cross section was less than 10%, the MC area ratios in other sites of the tumor showed only a small variance, but in the patients with an MC area ratio of 10% or more, a large variance was observed according to the site. The prognosis for patients with an area ratio of 10% or more was less favorable than that for patients with no MC and with an MC area ratio of less than 10% (P less than 0.05). These results suggest that it is reasonable to handle the patients with an MC area ratio of 10% or more as a group. PMID- 2547508 TI - Radical pancreatectomy for ductal cell carcinoma of the head of the pancreas. AB - Seventy-four patients were treated with a radical or a nonradical pancreatectomy for ductal cell carcinoma of the head of the pancreas. Their survival rates and the selection of the operative procedure were evaluated. In 32 patients, a radical pancreatectomy was attempted where there was sufficient clearance of regional or juxta-regional lymph nodes beyond the group of suspected metastatic nodes, as well as a resection of a greater margin of soft tissue around the pancreas. These patients' cumulative 5-year survival rate was 33.4%. In 14 Stage I or Stage II patients, the cumulative 5-year survival rate was 46.4%. In 18 Stage III or Stage IV patients, the cumulative 5-year survival rate was 20.7%. For 42 patients treated with a nonradical pancreatectomy with the dissection of lymph nodes adjacent to the pancreas or of regional lymph nodes but with insufficient clearance of the soft tissue around the pancreas, the cumulative 2 year and 3-year survival rates were 5.4% and 0%, respectively. In seven patients with Stage II carcinoma, the survival rate was 16.7% after 2 years and 0% after three years. In 35 Stage III or Stage IV patients, the survival rate was 3.2% after 2 years and 0% after 3 years. Thus, the survival rates were significantly higher in patients treated with radical operation than in patients who had nonradical operation. These results indicate that a radical pancreatectomy with sufficient lymph node clearance with the surrounding connective tissue around the pancreas is indispensable to cure patients with ductal cell carcinoma of the pancreas. PMID- 2547510 TI - A hypodiploid karyotype found in immortal human cells selected from a wide spectrum of posttransformation chromosomal complements. AB - A simian virus 40 (SV40) DNA fragment, encompassing the whole early region but having a defective origin of DNA replication, was previously used to transform human fibroblast cells derived from a patient suffering from xeroderma pigmentosum complementation group C (XP-C). Two independent SV40 transformants had acquired immortality in culture. Unlike most SV40-transformed human fibroblasts, the two established XP-C cell lines possessed an identical hypodiploid karyotype of 44,XX,-19,Xq+,-22,15p+. We now show that prior to immortalization the two SV40 transformants display a very wide spectrum of karyotypes with regard to chromosome number. A similar variety of chromosomal complements is present in four independent mortal SV40 transformants of the same parental XP-C cell line as well as in a mortal SV40-transformed xeroderma pigmentosum group D cell line. The rarity of the immortalization event, coupled with the coincident occurrence of identical karyotypes in the two immortal cell lines, suggests that the immortal lines arose through selection of a peculiar karyotype from among those of the parent SV40-transformed fibroblasts, and that this peculiar hypodiploid karyotype may be related to, and perhaps even necessary for, the establishment of immortality. PMID- 2547509 TI - A case-control study of risk factors for hepatoblastoma. A report from the Childrens Cancer Study Group. AB - Parents of 75 children with hepatoblastoma, registered with the Childrens Cancer Study Group, and 75 age-matched controls, who had been identified through random digit dialing were interviewed. No evidence was found to support the primary study hypotheses relating to hepatitis infection, maternal estrogen exposure, alcohol consumption, smoking, or potential sources of nitrosamines. Case mothers were more likely to report occupational exposure to metals (odds ratio [OR] = 8.0, P = 0.01), petroleum products (OR = 3.7, P = 0.03), and paints or pigments (OR = 3.7, P = 0.05). Metal exposures were commonly to welding or soldering fumes, and most occurred daily, before and during the index pregnancy. Petroleum product exposures were predominantly to lubricating oils or protective greases. The only significant paternal exposure was to metals (OR = 3.0, P = 0.01) and the risk with exposure to petroleum products was marginally significant (OR = 1.9, P = 0.06). These findings provide further evidence that occupational exposures may increase the risk of cancer in offspring. PMID- 2547511 TI - Involvement of c-myc oncogene in lymphoma cell lines with no detectable chromosome rearrangement of band 8q24. AB - Two lymphoma cell lines of B-cell type were established from Japanese patients with diffuse small noncleaved cell lymphoma. Cytogenetic analysis revealed a 14q+ marker chromosome in both cell lines, and a t(8;14)(q24.1;q32.3) seemed most likely to have occurred. The chromosome 8 pair, however, had no abnormalities. Molecular analysis demonstrated c-myc amplification lacking gross rearrangement in one cell line and genetic rearrangement of c-myc at the first intron as well as aberrant sizes of c-myc mRNA in the other cell line. In the latter case, it is possible that a t(8;14)(q24.1;q32.3) was buried in an unrecognized complex translocation. A combination of cytogenetic and molecular studies to determine the precise nature of the 14q32 translocation is discussed. PMID- 2547512 TI - Papillomaviruses in anogenital cancer as a model to understand the role of viruses in human cancers. AB - Infections with specific types of human papillomaviruses (HPV) have emerged as necessary but not sufficient factors for the development, at least, of the majority of cervical, vulvar, penile, and perianal cancers. Evidence has accumulated for their causal role in the induction of anogenital premalignant lesions. Genetic events underlying the mechanism of anogenital carcinogenesis have become increasingly understood. A host cell-mediated intracellular control down-regulating specific HPV genes (E6, E7) in replicating normal cells appears to be interrupted in cancer cells, probably due to structural modifications of the respective host cell genes acquired in the course of HPV DNA persistence. Since genital HPV infections are ubiquitous, cofactors which modify controlling host cell genes are likely to determine the different geographic rates of cervical cancer incidence. PMID- 2547514 TI - Alkaline phosphatase isozymes in human testicular germ cell tumors, their precancerous stage, and three related cell lines. AB - The four known isozymes of the human alkaline phosphatase (ALP) were detected by isoelectric focusing in extracts of various types of germ cell tumors, three related cell lines, and their precancerous elements (atypical germ cells). In seminoma, placental alkaline phosphatase (PLAP) and germ cell alkaline phosphatase (PLAP-like) could be separated by isoelectric focusing following isolation by immunoaffinity. The occurrence of both isozymes in seminoma could explain partial heat sensitivity and variation in electrophoretic patterns of the seminoma isozyme frequently observed upon starch gels, in comparison to the normal placental phenotype. The four ALP isozymes are produced not only in germ cell tumors, but already in precancerous tissues. Quantitative analysis showed that the amount of the four isozymes varies in parallel in the tumors tested. Maximal expression was found in seminoma. The relation between ALP gene overexpression and gene amplification by polyploidy of chromosomes 1 and 2 in these lesions is discussed. On the other hand, the ectopic expression of intestinal alkaline phosphatase and PLAP associated with overexpression of PLAP like in tumor cells as well as in their precancerous stage indicates gene activation by some unknown mechanisms, probably a regulatory process affecting the three tissue-specific ALP genes simultaneously. PMID- 2547513 TI - ras oncogenes in human cancer: a review. AB - Mutations in codon 12, 13, or 61 of one of the three ras genes, H-ras, K-ras, and N-ras, convert these genes into active oncogenes. Rapid assays for the detection of these point mutations have been developed recently and used to investigate the role mutated ras genes play in the pathogenesis of human tumors. It appeared that ras gene mutations can be found in a variety of tumor types, although the incidence varies greatly. The highest incidences are found in adenocarcinomas of the pancreas (90%), the colon (50%), and the lung (30%); in thyroid tumors (50%); and in myeloid leukemia (30%). For some tumor types a relationship may exist between the presence of a ras mutation and clinical or histopathological features of the tumor. There is some evidence that environmental agents may be involved in the induction of the mutations. PMID- 2547515 TI - High affinity binding of estrogen receptor to recombinant plasmids containing (dA dC)n.(dG-dT)n sequences. AB - Estrogen receptor is a gene regulatory protein that is present in a subset of breast tumors and in normal sex accessory tissues of vertebrates. The receptor protein forms a high affinity complex with estradiol and mediates its action. The mechanism of action of estrogen receptor (ER) involves its binding to specific DNA sequences in the genome and the stimulation of the expression of estrogen responsive genes. In order to understand the molecular characteristics of ER-DNA interactions, we studied the relative binding affinity of rabbit uterine ER to recombinant plasmids: pDHf2 with 23-base pair (dA-dC)n.(dG-dT)n insert; pDHf14 with 60-base pair (dA-dC)n.(dG-dT)n insert; pDHg16 with 23-base pair (dG-dC)n.(dG dC)n insert; and the parental plasmid pDPL6 without any insert. We conducted this study by using a DNA-cellulose elution assay in which the receptor was bound to DNA-cellulose and then eluted with different concentrations of plasmid DNAs in 10 mM Tris.HCl and 1 mM dithiothreitol, pH 7.4. The efficacy of a plasmid to bind to ER was determined from the concentration of the plasmid DNA required to elute 50% of the receptor from DNA-cellulose. The 50% elution concentrations were as follows: pDHf2, 3.2 micrograms/ml; pDHf14, 0.85 micrograms/ml; pDHg16, 200 micrograms/ml; and pDPL6, 340 micrograms/ml. Linearization of pDHf14 with HindIII reduced its binding efficacy only slightly, with a 50% elution concentration of the linear plasmid at 3.2 micrograms/ml. In contrast, the linear polynucleotide, poly(deoxyadenylate-deoxycytidylate).poly(deoxyguanylate-deoxythymidylat e [poly(dA-dC).poly(dG-dT)] and calf thymus DNA were 1650 and 880-fold less efficient, respectively, than pDHf14 in eluting ER from DNA-cellulose. These results show that the presence of short stretches of (dA-dC)n.(dG-dT)n sequences has a dramatic effect in increasing the binding affinity of ER to DNA. Since these sequences are reported to be conformationally ultrapolymorphic, a possible explanation for this increased binding might be the assumption of a conformational state of the inserted sequence to a form that has high affinity for ER. PMID- 2547516 TI - Dependence of etoposide-induced cytotoxicity and topoisomerase II-mediated DNA strand breakage on the intracellular ionic environment. AB - We have found that blockade of the Na+,K+-pump by the cardiac glycoside ouabain protects human A549 and hamster V79 cells from the cytotoxic effects of the topoisomerase II poison etoposide. One thousand-fold higher concentrations of ouabain were required to protect V79 cells compared to A549 cells. Since this difference parallels previously measured differences in pump sensitivity, it suggests that protection is mediated directly through pump blockade. Ouabain affected neither the cellular influx nor efflux of etoposide. However, pump blockade did decrease the formation of etoposide-induced DNA-topoisomerase, II cleavable complexes, assessed as single and double strand DNA breaks using alkaline and neutral elution. To determine if this decrease were a direct effect of change in ionic environment produced by pump blockade, experiments with isolated nuclei and partially purified topoisomerase II were performed. Etoposide induced cleavable complex formation and topoisomerase-mediated decatenation were assessed in buffers which mimicked either normal intracellular ionic conditions or those produced by ouabain. Compared to the buffer which resembled the normal intracellular ionic conditions, the buffer that mimicked the conditions produced by pump blockade produced fewer etoposide-mediated cleavable complexes in isolated nuclei and less decatenating activity of partially purified topoisomerase II. These findings demonstrate that inhibition of the Na+,K+-pump causes an alteration in the intracellular ionic environment which decreases the activity of topoisomerase II, thus producing a decrease in etoposide-induced cleavable complex formation and cytotoxicity. Since ionic changes occur inside normal cells during progression through the cell cycle as well as in cells that have undergone transformation, these data suggest that the intracellular ionic environment plays a role in determining the sensitivity of normal and malignant cells to this group of chemotherapeutic agents. PMID- 2547517 TI - Chemosensitivity and radiosensitivity of small cell lung cancer cell lines studied by a newly developed 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) hybrid assay. AB - The 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) hybrid assay was developed by technically combining the human tumor clonogenic assay and the MTT assay to make the most of both assays. This assay was able to estimate the in vitro growth of cultured cell lines and of tumor cells in pleural effusion, suggesting the possibility of its use for assessment of chemosensitivity and radiosensitivity of fresh tumor samples. Multiple cell lines [including morphological and/or phenotypical in vitro converters and cisplatin (CDDP)-resistant lines] were established from three patients with small cell lung cancer at different stages of the disease. Chemosensitivity of these cell lines to four commonly used chemotherapeutic drugs was tested by the MTT hybrid assay. SK1 and SK3 lines were established from Patient S. K. before and after chemotherapy and radiotherapy, respectively. SK3/CDDP, a CDDP-resistant line derived from the SK3 line, was 30-fold more resistant to CDDP [50% inhibiting dose (IC50), 21.5 micrograms/ml] than the SK1 line. In Patient M. O., MOA2/CDDP, a CDDP-resistant line derived from MOA2 (an in vitro converter from the MO line), was 41-fold more resistant to CDDP (IC50, 37 micrograms/ml) than the parent MO line. From Patient T. M., TM1 and TM2 lines were established before and after chemotherapy, respectively. The latter showed 6-fold more resistance to CDDP than the former. Chemosensitivity of these lines to three other drugs, 4 hydroperoxycyclophosphamide, Adriamycin, and etoposide, suggested cross resistance between CDDP and 4-hydroperoxycyclophosphamide. Radiosensitivity study was also carried out with the MTT hybrid assay. The MOA2 line was more resistant [Do, 3.0 Gy; extrapolation number (n), 4.0] than the parental MO line (Do, 1.6 Gy; n, 2.1). There was no clear difference in radiosensitivity between the cell lines established before and after radiation therapy in Patient S. K. PMID- 2547518 TI - Relationships among cell survival, O6-alkylguanine-DNA alkyltransferase activity, and reactivation of methylated adenovirus 5 and herpes simplex virus type 1 in human melanoma cell lines. AB - O6-Alkylguanine-DNA alkyltransferase (ATase) activity and host cell reactivation (HCR) of 5-(3-methyl-1-triazeno)imidazole-4-carboxamide (MTIC)-methylated viruses were compared in human melanoma cell lines that were sensitive or resistant to killing by the antitumor DNA-methylating agent MTIC. Enhanced HCR of adenovirus 5 (defined as the Mer+ phenotype) generally showed a semiquantitative correlation with the natural or induced resistance of the host cells to the toxic effects of MTIC and to the level of ATase activity. However, one MTIC-resistant cell line was found (MM170) which had a low level of ATase and intermediate HCR of adenovirus. The HCR of herpes simplex virus type 1 (HSV-1) was enhanced in the Mer+ cells that had natural resistance to MTIC compared with Mer- cells. On the other hand, HCR of HSV-1 in Mer+ cells with induced resistance to MTIC was similar to that in Mer- cells. Neither adenovirus 5 nor HSV-1 infection induced ATase activity in Mer- cells. This indicates that resistance to the toxic effects of methylating agents is not invariably associated with high levels of ATase activity in human melanoma cells. Furthermore, while induction of the Mer+ phenotype from Mer- cells was usually accompanied by the recovery of ATase activity, induced Mer+ cells had less proficient repair than natural Mer+ cells, as judged quantitatively by slightly lower cellular resistance and qualitatively by deficient HCR response for HSV-1. These results suggest that the Mer- and induced Mer+ cells lack an ATase-independent DNA repair mechanism. No differences in MTIC-induced DNA repair synthesis or strand breaks were found between the Mer , natural Mer+, and induced Mer+ phenotypes. However, UV-induced DNA repair synthesis was higher in the natural Mer+ than in the Mer- or induced Mer+ cells, both of which had increased cellular sensitivity to the antimetabolites methotrexate and hydroxyurea. These differences may be related to the effects observed with MTIC. A wide range of ATase activities was found in human melanoma biopsy material. PMID- 2547520 TI - Relationship between cell membrane potential and natural killer cell cytolysis in human hepatocellular carcinoma cells. AB - One of the body's natural defense mechanisms against tumor cells is lysis of the invading cell by cytotoxic T-cells and natural killer (NK) cells. Five human hepatocellular carcinoma cell lines were found to have different sensitivities to killing by peripheral blood monocytes in a 51Cr release assay. This killing was demonstrated to be due to NK cell lysis. Electrical recording measurements of the membrane potentials of these five cell lines showed different values for each line, all below values reported for normal hepatocytes. Correlation between mean cell membrane potential, and sensitivity to NK lysis, revealed an inverse relationship. In this study we demonstrate that the lower the mean membrane potential of a human hepatocellular carcinoma cell line, the more sensitive it is to NK cell cytolysis. Cell surface positive potential did not correlate with NK cytolysis and only a weak correlation was found between cell membrane negative potential and cell surface positive potential between cell lines. PMID- 2547519 TI - Directed migration of murine and human tumor cells to collagenases and other proteases. AB - Tumor cell motility and the passage of tumor cells through various tissue matrices, including basement membrane, are important components of the metastatic process. Proteolytic enzymes, including a type IV collagen-specific collagenase, have been demonstrated to play a significant role in extracellular matrix and basement membrane degradation. In addition, exogenous collagenase has been shown to enhance the motility of some tumor cells independent of its effect on collagen containing material. Previous studies have also indicated that collagen fragments are chemotactic for many tumor cells. We therefore studied the effect of type I and type IV collagen-specific collagenases, other enzymes involved in collagenase activation and connective tissue degradation, and subsequent collagen degradation products on the directed migration of tumor cells. We report that type I and type IV collagen-specific mammalian collagenases were potent chemoattractants as were native type I and type IV collagens and collagen fragments. Collagenase inhibitor SC44483 inhibited the type IV collagenase-stimulated migration. Collagenase pretreatment of the tumor cells potentiated the migratory response of the tumor cells to collagen and collagen fragments. The plasminogen activator, urokinase, as well as plasminogen itself also enhanced the directed migration of tumor cells in concentrations that suggest involvement of the appropriate cell surface receptor. The chemotactic response of tumor cells to the proteases studied extends the prior report of a role for collagenases and other matrix-active enzymes in tumor cell behavior in addition to matrix degradation. PMID- 2547521 TI - Smoking and infectious agents and risk of in situ cervical cancer in Sydney, Australia. AB - In a study of 116 in situ cervical cancer patients and 193 matched community controls in Sydney, Australia, smoking was found to be a major risk factor. Current smokers had a adjusted relative risk [RR] of 4.5 compared to nonsmokers [95% confidence interval (CI) 2.2-9.1] and exsmokers a RR of 1.3 [95% CI 0.6 3.0]. There was a stepwise dose-response relationship between risk and number of cigarettes smoked (30+ cigarettes/day, RR = 5.1, 95% CI 1.5-17.3); this dose response relationship was more marked among current smokers. Years of cigarette smoking was not consistently related to risk. Exposures to herpes simplex virus type 2 and cytomegalovirus as measured by antibody prevalence were unrelated to risk (RR = 1.1 for both measures). However, cases appeared to have more exposure than controls to herpes simplex virus type 1 (RR = 2.3, 95% CI 1.1-4.4). PMID- 2547522 TI - Glioblastoma multiforme masquerading as a pleomorphic xanthoastrocytoma. PMID- 2547523 TI - Stereotactic interstitial irradiation of diencephalic tumors with iridium 192 and iodine 125: 10 years follow-up and comparison with other treatments. AB - The only possible treatment of non-removable tumors of the diencephalon or recurring tumors invading the diencephalon after partial resection or percutaneous radiotherapy is interstitial irradiation (Curie therapy). With the CT/MRI stereotactic method, biopsy for histological tumor classification can be performed and 125I or 192Ir implanted, provided the neuroimaging methods show the delimitation of the tumor, its diameter does not exceed 3 cm and, given the patient's condition, focal irradiation seems advisable. As of 31 August 1987, a total of 1883 cases had been stereotactically biopsied and interstitially irradiated. The indications and results are reported in 204 patients under 18 years of age with diencephalic and deep-seated astrocytomas, after a follow-up period of up to 10 years; the data were compared for tumor resection, percutaneous irradiation, and for interstitial irradiation. The latter proved to be the most effective treatment. PMID- 2547524 TI - Role of the octamer motif in hybrid cell extinction of immunoglobulin gene expression: extinction is dominant in a two enhancer system. AB - We have shown previously that genes activated by the immunoglobulin heavy chain (IgH) enhancer or promoter in mouse myeloma cells are extinguished upon fusion of the myeloma with a mouse T cell lymphoma. Here we show that the conserved octamer sequence shared by the IgH enhancer and promoter, when multimerized to form a tissue-specific enhancer, can also render a gene extinguishable under the same experimental conditions. Extinction, however, is not correlated with either absence of the tissue-specific transcription factor OTF-2 or loss of its ability to bind the octamer sequence. It was also found that extinction mediated by the IgH enhancer is dominant to transcriptional activation by the SV40 enhancer. We propose, therefore, that the T cell-negative regulator responsible for IgH gene extinction does not simply prevent IgH enhancer activation but interferes with gene expression more directly, perhaps by disrupting the transcription complex established as a result of tissue-specific enhancer activation. PMID- 2547525 TI - The Epstein-Barr virus origin of plasmid replication, oriP, contains both the initiation and termination sites of DNA replication. AB - Epstein-Barr virus (EBV) oriP contains two components, a dyad symmetry element and a direct repeat element, that, in the presence of EBV nuclear antigen 1, are necessary and sufficient for plasmid replication. We have examined the replicative forms generated by EBV oriP using 2D gel electrophoresis. The patterns obtained from an oriP plasmid in a transfected cell line indicate that the site of initiation of DNA replication is at or very near the dyad symmetry element, while the direct repeats contain a replication fork barrier and the termination site. Thus, replication from oriP proceeds in a predominantly undirectional manner. The patterns obtained from cells immortalized by EBV suggest that replication from oriP proceeds similarly in the viral genome. PMID- 2547526 TI - Leg weakness associated with Powassan virus infection--Ontario. PMID- 2547528 TI - Intensive chemotherapy with autologous bone marrow transplantation for small-cell lung cancer. AB - Since 1980, 75 patients with small-cell lung cancer (SCLC) have been entered into four consecutive studies of high-dose chemotherapy using autologous bone marrow transplantation (ABMT) to assist haematological recovery. In the first study, 25 patients were treated with cyclophosphamide (160-200 mg/kg) as the sole chemotherapy; in the second (26 patients), the cycle of high-dose cyclophosphamide (with or without 800-1,200 mg/m2 etoposide) was repeated as induction treatment. In the first study, response was high [14 complete responses (CR), 7 partial responses (PR)] but was not increased by repeating the cycle (15 CR, 8 PR), and survival was slightly worse in the second trial. In the third study, 15 patients were treated with doxorubicin, vincristine and etoposide for two cycles and then with 200 mg/kg cyclophosphamide. Although high-dose cyclophosphamide increased the complete response rate, the additional responses were short-lived. In the final study, an attempt was made to increase the initial CR rate by combination chemotherapy using carboplatin (400-600 mg/m2), etoposide (120 mg/m2 x 4) and either high-dose cyclophosphamide (40 mg/kg x 4) or melphalan (140 mg/m2). Although all nine patients responded, none underwent a CR. The long term survival (up to 7 years) does not appear to be different from that in comparably selected cases treated with conventional chemotherapy. PMID- 2547529 TI - A phase II study of ifosfamide in the treatment of relapses in Wilms' tumor. AB - The purpose of this study was to evaluate the antitumor activity and tolerability of ifosfamide (IFO) at a dose of 3 g/m2, given on 2 consecutive days every 2 weeks, in advanced Wilms' tumor patients in whom conventional therapy had failed. Mesna and hyperhydration were concomitantly given to minimize bladder toxicity. A total of 21 patients with advanced Wilms' tumor were entered in the study. The response observed after two courses was complete in 6 patients and partial in 5; 10 did not respond; the median duration of response was 2 months (range, 1-7 months). Leucopenia caused a delay in therapy for 1 or 2 weeks in only three cases. Neither fever nor infection were observed. Of 7 patients who developed hematuria, 3 were among the 17 who concurrently received mesna. The urotoxicity did not interfere with subsequent therapy in these three cases. PMID- 2547527 TI - Identification of anthracyclines and related agents that retain preferential activity over adriamycin in multidrug-resistant cell lines, and further resistance modification by verapamil and cyclosporin A. AB - A range of anthracyclines and related compounds were evaluated for activity against murine and human cell lines exhibiting multidrug resistance (MDR). Cell lines used were the NCI-H69 human small-cell lung cancer line and the EMT6 murine mammary tumour line, together with their multidrug-resistant counterparts produced by in vitro exposure to Adriamycin (ADM). Chemosensitivity testing was carried out using the tetrazolium (MTT) dye assay. Results were expressed as the ratio of the ID50 for the resistant line to that obtained in the parent, i.e. the resistance factor (RF). Compounds exhibiting much lower RF values than ADM in both resistant cell lines were identified as those anthracyclines with 9-alkyl substituents and those with certain changes to the amino sugar residue at position 3' and 4', together with the anthracenedione mitoxantrone (MIT). In a further attempt to overcome resistance, we used four of these compounds, Ro 31 1215, 4'-deoxy-4'-iodo-ADM (iodo-ADM), aclacinomycin A (ACL) and MIT (all yielding low RF values), in combination with the resistance modifiers verapamil (VRP) and cyclosporin A (CYA). Additional enhancement of chemosensitivity was achieved in the ADM-resistant sublines, as shown by the further decrease in RF values. At the concentrations used, the largest effects were generally seen with CYA, and the combination of this modifier with ACL and MIT was particularly effective. For the H69/LX4 resistant line, the latter combinations gave RF values approaching unity. These findings point to the use of analogues with the 9-alkyl substituent and/or specific changes to the sugar residue in combination with resistance modifiers as a therapeutic strategy for circumvention of the MDR phenotype. PMID- 2547530 TI - Combination chemotherapy in malignant non-seminomatous germ-cell tumors: results of a cooperative study of the German Society of Pediatric Oncology (MAKEI 83). AB - In January 1983, the German Society of Pediatric Oncology started a cooperative trial (MAKEI 83) for non-testicular germ-cell tumors. The pilot phase closed in December 1985. The treatment regimen was stratified according to histology, tumor site and tumor stage. In malignant non-seminomatous germ-cell tumors (mNSGCTs), chemotherapy consisted of four courses of 3 mg/m2 vinblastine, on days 1 and 2 and 15 mg/m2 bleomycin on days 1-3, given by continuous infusion, and 20 mg/m2 cisplatin on days 4-8 with mannitol diuresis. Courses were repeated every 3 weeks. In mNSGCT patients with ovarian FIGO stages III-IV or extragonadal primaries, second-look surgery was carried out, followed by four additional courses of chemotherapy with 100 mg/m2 VP-16 on days 1-3, 1.5 g/m2 ifosfamide on days 1-5 with mesna uroprotection and 20 mg/m2 cisplatin on days 1-5 with mannitol diuresis. In patients with sacrococcygeal germ-cell tumors, en bloc resection of the tumor, including the coccygeal bone, was mandatory. During the registration period, 57 patients with mNSGCTs were entered: 37 protocol patients and 20 follow-up patients. The event-free survival for protocol patients at 57 months was 78% +/- 6% and that for follow-up patients was 40% +/- 10% (Kaplan Meier): the crude survival for both groups was 83% +/- 6% and 54% +/- 12%, respectively. After a review by a panel of pathologists, the histological diagnoses in 7% of all registered cases of germ-cell tumors were changed. The results of the present studies show that the histological subclassification of mNSGCTs, tumor site and tumor stage no longer had prognostic value. PMID- 2547531 TI - Depressed activities of purine enzymes in lymphocytes of patients infected with human immunodeficiency virus. AB - Enzyme activities were studied in peripheral blood lymphocytes from patients infected with, or at risk for, infection with human immunodeficiency virus (HIV). No significant differences were observed in the HIV-infected and HIV-seronegative high-risk patients with regard to enzyme activities of hypoxanthine-guanine phosphoribosyltransferase (EC 2.4.2.8) and purine nucleoside phosphorylase (EC 2.4.2.1) in peripheral blood. Adenosine deaminase (EC 3.5.4.4) was significantly (P less than 0.02) depressed in asymptomatic HIV-seropositive patients and HIV seronegative patients at high risk of HIV infection as compared with a healthy HIV-seronegative population. Adenosine kinase (AK, EC 2.7.1.20) was significantly increased in the asymptomatic seropositive (P less than 0.02) and also in the HIV seronegative high-risk groups (P = 0.01) compared with the normal controls. AK activity was significantly lower in subjects with AIDS than in the asymptomatic (P less than 0.002) and high-risk groups (P less than 0.01). Taken together, these results indicate that adenosine deaminase and AK activities are influenced by the health of the patient, and that measurement of AK activity may prove useful in monitoring the clinical progress of patients with HIV infection. PMID- 2547532 TI - Influence of blood pressure, heart rate, age, and sex on concentrations of atrial natriuretic factor and cyclic GMP in 124 volunteers. AB - The significance of increased atrial natriuretic factor (ANF) in relation to blood pressure and age is still controversial. We investigated the influence of blood pressure, age, and some other variables on ANF and its putative second messenger, cGMP. Samples for ANF and cGMP detection were taken from 124 ostensibly healthy individuals who were donating blood. Samples were also collected from 27 volunteers before and after blood donation, to study the influence of bleeding. During blood donation, ANF increased from 78.9 to 87.4 ng/L (P = 0.0035), whereas cGMP remained unchanged. ANF concentrations in 124 healthy individuals, corrected for the influence of bleeding, were 61.5 (SD 26.1) ng/L, with a 95% confidence interval of 10.0 to 112.1 ng/L. Mean cGMP concentrations in plasma were 2.9 (SD 1.45) nmol/L, with a 95% confidence interval of 0.4 to 5.75 nmol/L. Multivariance analysis revealed no significant influence of blood pressure, age, heart rate, or sex on concentrations of either ANF or cGMP in plasma. PMID- 2547533 TI - Use of molecular probes to detect human cytomegalovirus and human immunodeficiency virus. AB - Human cytomegalovirus (HCMV) and human immunodeficiency virus (HIV) cause severe disease. The identification of these viruses in clinical specimens and understanding the progression of infection and diseases relating to HCMV and HIV are essential to develop effective means for treatment and prevention. Here we describe the application of molecular probes to the diagnosis and pathogenesis of HCMV and HIV. In situ hybridization and the amplification procedure of polymerase chain reaction are used to detect both viruses; these techniques have provided important information regarding the pathogenesis of HCMV and HIV. A new technique, target cycling, may also prove useful for the detection of viruses by enriching for target sequences. The continued application of molecular probes to pathogenetic studies of HCMV and HIV promises to further our knowledge of these viruses, and of their interaction. PMID- 2547535 TI - Coxsackie virus and urogenital pathology. AB - In 43 women (average age 49.6 years), and 70 men (average age 55.2 years) with pathology of the genito-urinary apparatus, seroantibodies to Coxsachie virus B were measured using the passive hemoagglutination method and the virus was isolated in the uterine. Viral isolation test was negative in all urine samples tested. Seropositivity for Coxsackie virus was reported in 26 women (60.46%) and in 51 men (72.85%). Positivity to B1 was 37.16% (42 cases), B2 in 38.05% (43 cases), and to B4 in 35.39% (40 cases). 17.69% (20 cases) of patients were seropositive to only one serotype, 17.69% (20 cases) to 5 serotypes, 14.15% (16 cases) to 3 serotypes, 9.73% (11 cases) to 2 serotypes, and 8.84% (10 cases) to 4 serotypes. B1, 2, 3, 4, 5 (25.97%; 20 cases) and B2, 3, 4, 5 (7.79%; 6 cases) were the most frequent associations. Seropositivity to Coxsackie virus was reported in 100% of patients (4 cases) with urethral caruncola, in 84.21% (16 cases out of 19) with cancer of the bladder, in 81.81% with cystitis (9 cases out of 11) and in 80% with prostatitis (8 out of 10 cases). In relation to sex, seropositivity was higher in males in cases of calculosis (75%; 9 cases out of 12 against 28.57%; 2 cases out of 7) and in cystitis (100%; 6 cases against 60%; 3 cases out of 5). Further studies are necessary to determine the clinical significance of serum Coxsackie virus antibodies in patients with urological pathology in the absence of urinary elimination of Coxsackie virus. PMID- 2547534 TI - Rapid radioimmunoassay of circulating chromogranin A: in vitro stability, exploration of the neuroendocrine character of neoplasia, and assessment of the effects of organ failure. AB - Chromogranin A is a useful probe of neuroendocrine neoplasia in humans. Here we optimize a rapid, sensitive radioimmunoassay modification for detecting chromogranin A in humans and other species. The site of chromogranin A circulation is the acellular plasma; platelets contain no chromogranin A immunoreactivity. The immunoreactivity in plasma is stable to repeated freezing and thawing, prolonged incubation at 37 degrees C, and lyophilization. Venipuncture alone resulted in modest (+ 12%, P less than 0.03) increase in chromogranin A in plasma. Several classic neuroendocrine neoplasia pheochromocytoma, carcinoid tumor, neuroblastoma, and (vasoactive intestinal polypeptide)oma-produce markedly increased chromogranin A in plasma. By contrast, subjects with malignant melanoma, renal cell carcinoma, and thymoma all had normal values for chromogranin A. Hypersecretion of human choriogonadotropin beta subunit from both malignant (choriocarcinoma) and normal (placenta) syncytiotrophoblast cells was unaccompanied by an increase in chromogranin A, a dissociation compatible with the lack of granular storage and release of syncytiotrophoblastic peptide hormones. Both hepatic and renal failure resulted in increased chromogranin A in plasma, with renal failure leading to concentrations otherwise seen only in neuroendocrine neoplasia. These observations refine the diagnostic specificity of chromogranin A in plasma. PMID- 2547536 TI - Adenylate cyclase and cyclic AMP-phosphodiesterase activities of fetal membranes: effect of insulin. AB - Basal activity and response to insulin of adenylate cyclase and cyclic AMP phosphodiesterase were measured in the fetal membranes before and after labor. Basal activity of adenylate cyclase showed only slight variations after delivery when compared to that observed before labor. Enzyme activity was significantly although weakly decreased by insulin both in the amnion and in chorion before labor, while after delivery a weaker non-significant inhibition was observed. Basal activity of cyclic AMP-phosphodiesterase was lower after delivery with respect to that observed before labor. Insulin inhibited enzyme activity in all conditions. Cyclic AMP levels in intact tissue were not substantially modified by insulin. PMID- 2547537 TI - One step sandwich enzyme immunoassay for human type IV collagen using monoclonal antibodies. AB - Monoclonal antibodies were used in one step sandwich enzyme immunoassay (one step sandwich EIA) for human serum immunoreactive type IV collagen. The one step sandwich EIA using either polystyrene ball or microplate was characterized by carrying out two immunoreactions simultaneously, type IV collagen reacting with both a monoclonal antibody as a solid phase and a horseradish peroxidase-labeled monoclonal antibody (Fab') against human type IV collagen as a conjugate. Sensitivity of one step sandwich EIA system by using either polystyrene ball or microplate was 0.22 ng per tube or 0.04 ng per well for type IV collagen, and linearity was obtained between 0.22-40 ng/tube or 0.04-20 ng per well, respectively. Both methods gave reproducible quantitative analysis of immunoreactive type IV collagen levels in the sera of patients with hepatocellular carcinoma and patients with liver cirrhosis, which were apparently higher than the levels in the sera of healthy subjects. Protein immunoblotting shows that the immunoreactive type IV collagen trapped in our present one step sandwich EIA system was not the 7-S and NC1 domains of type IV collagen. PMID- 2547539 TI - The interaction of interleukin 2 with its receptor in the generation of suppressor T cells in antigen-specific and antigen-nonspecific systems in vitro. AB - The role of interleukin-2 (IL-2) in the activation of suppressor T cells was investigated using the monoclonal antibody anti-Tac, which blocks the binding of IL-2 to the 55-kDa IL-2-binding peptide. The addition of anti-Tac during a preculture period inhibited the generation of Epstein-Barr virus (EBV)-induced suppressor T cells and of suppressor T cells induced in an antigen-specific system by a high antigen (sheep red blood cell) concentration. The cells activated by a short 2- or 7-day preculture period with EBV to become suppressor effectors were of the T8, Tac-positive phenotype. However, the T8-positive T cells obtained from peripheral blood mononuclear cells precultured with EBV for 14 days continued to manifest suppressor function, even though they were no longer Tac positive. In summary, our studies indicate that anti-Tac, by producing a functional blockade of human IL-2 receptors, inhibits the generation of suppressor T cells in antigen-specific as well as antigen-nonspecific systems and that cells that no longer express the Tac antigen may also function as suppressors. PMID- 2547538 TI - Autoantibodies to DNA topoisomerase II in cryptogenic fibrosing alveolitis and connective tissue disease. AB - Sera from patients with autoimmune lung and connective tissue diseases were investigated for antibodies to topoisomerase II. Anti-topoisomerase II antibodies were detected by ELISA in 37% of sera from patients with cryptogenic fibrosing alveolitis (CFA), in one (8%) case of sarcoidosis and in 31% of sera from systemic lupus erythematosus (SLE) patients. Sera from rheumatoid arthritis, juvenile rheumatoid arthritis, progressive systemic sclerosis, Sjogren's syndrome and undifferentiated connective tissue disease were negative. CFA and sarcoidosis sera strongly reacted in immunoblotting with a 170 kD protein, also recognized by rabbit antiserum to recombinant topoisomerase II, while SLE sera presented a weak reaction. Pre-adsorption with dsDNA dramatically decreased the topoisomerase II binding in ELISA by the most positive SLE serum, but did not affect the binding by the most positive CFA serum, thus indicating that anti-topoisomerase II reactivity of SLE sera is probably due either to cross-reacting antibodies or, in part, to minor DNA contamination of our enzyme preparation. The determination of DNA topoisomerase II relaxation activity, performed after incubation with antibody-positive sera, showed that only CFA sera precipitate enzymatic activity. The finding that CFA presents antibodies to an enzyme essential to cell survival stresses the role of autoimmunity in the pathogenesis of idiopathic pulmonary fibrosis. This may offer further insight into the cause of autoimmune disease and prove a valuable tool in the study of enzyme molecular biology. PMID- 2547541 TI - Clinical herpes zoster shortly following primary varicella in two HIV-infected children. PMID- 2547540 TI - The influence of intra-articular hyaluronic acid on PGE2 and cAMP of synovial fluid. AB - Prostaglandin (PG)E2, cyclic adenosine monophosphate (cAMP), white blood cells (WBC), total protein (TP), total complement activity (CH50) and beta-2 microglobulin (beta-2-m) were measured at baseline and after eight days in the synovial fluid (SF) of 16 patients affected with knee-joint effusion due to various arthropathies. The volume of SF was also calculated. Eight patients--4 with rheumatoid arthritis (RA), 2 with recurrent monoarthritis (RM) and 2 with osteoarthritis (OA) were randomly allocated to the treatment with intra-articular injection of Hyalgan (HA, Na-hyaluronate, 20 mg/2 ml), while eight patients having similar arthropathies--4 RA, 2 RM and 2 OA--were not treated (control group). In the patients treated with HA a significant reduction of SF volume (from 28.5 +/- 5.1 ml to 20.5 +/- 4.0 ml; p less than 0.02) and PGE2 (from 96.1 +/- 22.7 pg/ml to 66.2 +/- 14.5 pg/ml; p less than 0.05) was found, whereas cAMP concentration was significantly increased (from 4.5 +/- 0.7 pmol/ml to 7.2 +/- 1.2 pmol/ml; p less than 0.05). No significant variations were observed in the control group. Moreover, no differences in WBC count, TP and beta-2-m and CH50 were found in either group. These data could suggest an anti-inflammatory effect of HA that appears to be mediated by PG-inhibition as well as cAMP stimulation. PMID- 2547542 TI - Captopril inhibits ouabain-sensitive Na+/K+-ATPase. AB - Captopril has been reported to inhibit ouabain-sensitive Na+/K+-ATPase activity in erythrocyte membrane fragments. We investigated the effect of captopril on two physiological measures of Na+/K+ pump activity: 22Na+ efflux from human erythrocytes and K+-induced relaxation of rat tail artery segments. Captopril inhibited 22Na+ efflux from erythrocytes in a concentration-dependent fashion, with 50% inhibition of total 22Na+ efflux at a concentration of 4.8 X 10(-3) M. The inhibition produced by captopril (5 X 10(-3) M) and ouabain (10(-4) M) was not greater than that produced by ouabain alone (65.3 vs. 66.9%, respectively), and captopril inhibited 50% of ouabain-sensitive 22Na+ efflux at a concentration of 2.0 X 10(-3) M. Inhibition by captopril of ouabain-sensitive 22Na efflux was not explained by changes in intracellular sodium concentration, inhibition of angiotensin-converting enzyme or a sulfhydryl effect. Utilizing rat tail arteries pre-contracted with norepinephrine (NE) or serotonin (5HT) in K+-free solutions, we demonstrated dose-related inhibition of K+-induced relaxation by captopril (10(-6) to 10(-4) M). Concentrations above 10(-4) M did not significantly inhibit K+-induced relaxation but did decrease contractile responses to NE, although not to 5HT. Inhibition of K+-induced relaxation by captopril was not affected by saralasin, teprotide or indomethacin. We conclude that captopril can inhibit membrane Na+/K+-ATPase in intact red blood cells and vascular smooth muscle cells. The mechanism of pump suppression is uncertain, but inhibition of ATPase should be considered when high concentrations of captopril are employed in physiological studies. PMID- 2547543 TI - Fatty acid changes in liver choline and ethanolamine glycerophospholipids in aspirin-treated rats fed linoleate, gamma-linolenate and fish oil. AB - The effects of dietary linoleic acid, gamma-linolenic acid and marine fatty acids on the development of aspirin-induced gastric hemorrhage and the distribution of liver glycerophospholipid fatty acids in fat-deficient growing rats were studied. Aspirin (100 mg/day)-treated and nontreated rats were fed for 7 days, a mixed diet of 2.5% safflower oil and 7.5% hydrogenated coconut oil (SFO/HCO) or 7.5% fish oil (SFO/FO), or 2.5% gamma-linolenate concentrate and 7.5% fish oil (GLA/FO). Gastric hemorrhage was induced in animals by aspirin treatment to various extents. It was not affected by FO feeding, but was significantly alleviated by GLA feeding. Aspirin treatment reduced the proportions of 20:4n-6 in liver phosphatidylcholine. FO feeding (in SFO/FO and GLA/FO rats) further reduced the 20:4n-6 level and replaced it by n-3 fatty acids. GLA feeding, on the other hand, elevated the proportion of 20:4n-6. As a result, the reduction of 20:4n-6 by fish oil feeding, was less significant in GLA/FO rats than in SFO/FO rats. The degree of gastric hemorrhage appeared to relate negatively to the levels of 20:4n-6 in liver phosphatidylcholine, and to the sum of 20:4n-6 and 20:5n-3 when FO was included in the diet. It is suggested that long-chain polyunsaturated fatty acids (20:4n-6 and 20:5n-3) per se in addition to being precursors of prostaglandins, may also affect the development of gastric hemorrhage, possibly by modulating the permeability of cell membranes in the gastric mucosa. PMID- 2547544 TI - Computed tomographic appearances of treated metastatic testicular tumours. AB - Recent therapeutic advances have considerably improved survival rates in patients with testicular tumours. Computed tomography (CT) permits accurate staging and follow-up monitoring of these patients, and it is important, therefore, to be familiar with the evolution of abnormalities on CT after treatment. This study was undertaken to evaluate these appearances and consider their impact on management. Eighty-three patients with metastatic testicular neoplasms had serial CT scans as part of their management. The average age was 34 years (range 19-66 years) with a mean follow-up period of 30 months (range 3-97 months). Seventy three patients had nodal disease at some stage: 44% reverted to normal on treatment, with no recurrence, while 30% regressed but did not clear completely. Thirty-six patients had lung lesions: although most responded to treatment persistent abnormality was observed in 39% of cases. Nine patients had extranodal disease. A variable response to treatment is reflected by CT findings that contribute to the development of a follow-up strategy whereby successful therapy may be recognised and residual or recurrent disease confirmed. PMID- 2547545 TI - Benign cholangioma of the liver. AB - The ultrasonic, computed tomographic, magnetic resonance and angiographic appearances of a case of benign cholangioma of the liver are presented. They have not been previously described. PMID- 2547546 TI - Marker antibodies in scleroderma and polymyositis: clinical associations. AB - Sera of 34 patients with progressive systemic sclerosis and of 11 patients with polymyositis/dermatomyositis (PM/DM) were analyzed by the immunoblotting technique for the presence of marker antibodies. The presence of anti-centromere, anti-Topoisomerase-I (anti-Topo-I) and anti-Jo-1 antibodies was found to be highly specific for the CREST syndrome, diffuse scleroderma and PM/DM, respectively, but only of limited sensitivity (78, 44 and 45%, respectively). Anti-Topo-I positive diffuse scleroderma patients had a more severe disease (digital pitting scars and renal insufficiency) than anti-Topo-I negative diffuse scleroderma patients. Anti-Jo-1 was associated with interstitial lung disease. Longitudinal studies showed a constant antibody pattern. Our results confirm the clinical usefulness of these marker antibodies. PMID- 2547548 TI - [The solid-cystic pancreas tumor]. AB - Solid-cystic pancreas neoplasia is a rare condition mostly observed in young adults and women. We report three cases of this rare tumor. In all three instances the patients were young, slender women. The tumors were removed by total resection of the pancreas. In two cases these tumors were very large. In two cases we also observed criteria which could indicate that these tumors were malignant: the tumors had infiltrated the adjacent spleen and adjoining lymphnode or displayed a destroyed capsula. Until now, such infiltrative or malignant qualities have not been reported. That such criteria can be conclusively evaluated as an indication of malignancy, remains, however, unclear. PMID- 2547547 TI - Differences in the release of 5'-nucleotidase and alkaline phosphatase from plasma membrane of several cell types by PI-PLC. AB - 1. We have compared the effect of phosphatidyl inositol specific phospholipase C (PI-PLC) on the attachment of both 5'-nucleotidase and alkaline phosphatase to the liver plasma membrane from different species. 2. Our results demonstrate differences in the susceptibilities of both enzymes to PI-PLC treatment in relation to their origin. 3. These results were confirmed by immunoblotting using polyclonal anti-5'-nucleotidase antibodies. 4. In addition, in a single animal, susceptibility of both enzymes to PI-PLC treatment is different from one tissue to another. 5. The different percentages of released enzymes could be explained either by a polymorphism in the anchoring of these proteins at the cell surface membrane, or by a different steric hindrance or environment at the cleavage site itself. PMID- 2547549 TI - [Clinicopathologic analysis of condyloma acuminatum of the vulva in 165 cases]. AB - Condyloma acuminatum of the vulva was diagnosed in 165 cases, of which 117 were identified during mass screening and 48 discovered in the outpatient clinic. The gross appearance and histologic characteristics were described. The lesions were removed surgically in 5 cases, and treated with topical 5-FU ointment in 4. The remaining cases were treated by cryosurgery. A permanent cure was achieved in all cases. The authors emphasize that the importance of condyloma acuminata should not be overlooked on account of its close relationship to vulvar cancer. PMID- 2547550 TI - [An etiological study of acute hemorrhagic conjunctivitis in Beijing area in 1984]. AB - Acute hemorrhagic conjunctivitis (AHC) has been widespread in China since 1971. An etiological survey of the sporadic outbreak in Beijing was done in 1984. Conjunctival swabs and sera were collected at the Eye Clinic of Peking Union Medical College Hospital and 10 strains of virus were isolated from 46 cases by culture on the HeLa cell line. 8 strains were identified as Enterovirus 70 by homologous antiserum J 670/71 obtained from the California State Laboratory of Viral and Rickettsial Diseases. 2 other strains defied neutralization by this antiserum and were eventually identified as Adenovirus type 3 and type 7. 30 paired sera were examined for neutralization antibodies to EV 70 (J 670/71). Among these, 23 (77%) showed fourfold or higher rise in titer. Furthermore, neutralization antibodies to EV70 (J 670/71) and the EV70 isolated from 7 patients with AHC by paired sera also showed a fourfold or higher rise in antibody titer, indicating that EV70 was the etiologic agent responsible for the 1984 outbreak in Beijing. PMID- 2547552 TI - Brain tumors in familial adenomatous polyposis. AB - Familial adenomatous polyposis was always believed to be a colonic disease of genetic determination with a high risk of development of cancer of the large bowel. Over the years the list of extracolonic manifestations of this disease, both benign and malignant, has amplified. Brain tumors and, in particular, medulloblastoma have not become recognized as major malignant extracolonic manifestations of familial adenomatous polyposis. They are of particular significance because, unlike most of the other manifestations, they occur prior to or early in the development of the colonic manifestations of this disease. This report documents the investigation of 168 kindreds in The Cleveland Clinic Familial Adenomatous Polyposis Registry in a search for those at-risk individuals who developed brain tumors. PMID- 2547553 TI - Insulin receptor kinase defects in insulin-resistant tissues and their role in the pathogenesis of NIDDM. PMID- 2547551 TI - Protein phosphorylation and control of tick salivary gland function. AB - Tick salivary glands are controlled by nerves, dopamine being a neurotransmitter at the neuroeffector junction. Dopamine and cyclic AMP (cAMP) stimulate fluid secretion by isolated salivary glands. Dopamine activates an adenylate cyclase to increase intracellular cAMP within the female salivary glands. Phosphoproteins whose levels of phosphate are affected by cAMP-dependent protein kinase have been identified in subcellular fractions. Protein(s) phosphorylated by cAMP appears to activate protein phosphatase in the salivary glands. Another phosphorylation pathway appears to act through protein kinase C because of an ability of phorbol esters (known activators of protein kinase C) to stimulate the phosphorylation of proteins, and an ability of a peptide factor in tick brain to metabolize salivary gland phosphoinositides, an event that often precedes activation of protein kinase C. Because cAMP modulates brain-factor-stimulated formation of inositol phosphates (products of phosphoinositide breakdown) an interrelationship between the two pathways seems likely. Evidence of regulatory processes, including protein phosphorylation/dephosphorylation reactions, will provide a basis for helping assess the physiological significance of secretory products and the role of the salivary glands in disease transmission. PMID- 2547554 TI - [Change in the activity of sphingomyelinase and the level of expression of nuclear oncogenes in reversible inhibition of protein synthesis by cycloheximide]. PMID- 2547555 TI - [Post- and presynaptic mechanisms of restoring nerve-muscle transmission with tubocurarine after the effect of an organophosphorous inhibitor of acetylcholinesterase]. PMID- 2547557 TI - International Symposium on Nedocromil Sodium. Paris, 13-15 June, 1988. Proceedings. PMID- 2547556 TI - [Changes in the myocardium in encephalomyocarditis of pigs]. AB - The myocardial lesions of pigs (age from 2.5 to 3.5 month) with encephalomyocarditis virus infection are presented. The disease is caused by members of the genus Cardiovirus (family Picornaviridae). In Greece it was first observed 1987 in two stables of pig-breeders with a mortality of 80%. The macroscopic changes are characterized by a myocardial degeneration with greyish pale linear or round foci and by an accumulation of a clear red to yellow fluid in the pericardium and the pleural cavity. The most important histologic changes of the hearts are a diffuse or focal myocarditis with a severe infiltration of histiocytes, lymphocytes and plasma cells. Furthermore, degeneration and necrosis of cardiac muscle cells are observed. Electron microscopic investigations of the myocardial cells reveal mild mitochondrial edema, severe perinuclear and intranuclear edema, myelin figures, intra cisternal sequestrations of the rough endoplasmic reticulum, disorganization of the myofibrillar Z-line, and crystalloid viral structures between the mitochondria. Polyribosomes and accumulations of viral particles are frequently found in membrane bound cysts within myocardial cells and the capillaries. Furthermore, viral particles (single or in cysts) and crystalloid viral structures are observed in the cytoplasm of edematous endothelial cells. PMID- 2547558 TI - Background and objectives of the basic research programme for nedocromil sodium. PMID- 2547559 TI - Effect of nedocromil sodium on early and late phase responses to allergen challenge in the guinea-pig. AB - We have developed a model of asthma in conscious guinea-pigs in which inhalation challenge with specific allergen induces both early and late phase reductions in specific airways conductance. Analysis of cells removed from the airways by bronchoalveolar lavage showed the presence of a neutrophilia, which reached a maximum at 17 hours, and an eosinophilia which developed more slowly, still increasing at 72 hours. Nedocromil sodium inhaled before challenge inhibited both the early and late phase responses. In contrast, the beta-adrenoceptor stimulant, salbutamol, inhibited only the early phase. When inhaled 6 hours after challenge, i.e. between the early and late phase responses, the late phase bronchoconstriction was prevented by nedocromil sodium but only partially by salbutamol. Evidence that the neutrophilia was not functionally associated with the late response was gained from the observations that it was inhibited by both nedocromil sodium and salbutamol, whereas only nedocromil sodium blocked the late phase airways response. When administered 6 hours after challenge, nedocromil sodium reduced eosinophil accumulation at 72 hours in parallel with inhibiting a secondary late response at this time. These results demonstrate that nedocromil sodium is able to prevent both early and late phase reductions in airways function in an animal model of allergic asthma. Whereas inhibition of the early response may reflect an effect on mast cell mediator release, the effects of nedocromil sodium on the late response and on eosinophil accumulation are strongly suggestive of an anti-inflammatory effect within the lung. PMID- 2547560 TI - Effect of nedocromil sodium on the bronchoconstrictor effect of neurokinin A in asthmatics. AB - In a double-blind, crossover study we have investigated the effect of nedocromil sodium on neurokinin A (NKA)-induced bronchoconstriction in asthmatics. 12 patients with mild asthma inhaled either nedocromil sodium 4mg or placebo, on 2 separate days, as 2 puffs from a metered-dose aerosol 30 minutes before challenge with NKA. On the placebo treatment day, NKA produced a concentration-dependent decrease in basal specific airway conductance (sGaw) and forced expiratory volume in 1 second (FEV1). The inhalation of nedocromil sodium 4mg inhibited the decrease in both sGaw and FEV1. The maximal percentage decrease in sGaw on the nedocromil sodium day was 27.0 +/- 5.2 (vs placebo, 53.3 +/- 5.4; p less than 0.05) and the maximal percentage decrease in FEV1 5.5 +/- 1.4 (vs placebo, 12.4 +/- 2.3; p less than 0.05). We conclude that nedocromil sodium protects against NKA-induced bronchoconstriction in asthmatics. PMID- 2547561 TI - Development of preclinical models for testing antiasthma drugs. AB - The antiasthmatic agent, sodium cromoglycate, owes its discovery to a series of antigen challenge tests carried out during the 1960s by an asthmatic, Roger Altounyan, on himself. Until recently, research efforts to identify new antiasthma drugs have relied heavily on screening methods which involved passively-sensitised mast cells. In theory these tests, such as rat passive cutaneous anaphylaxis, appeared relevant and showed sodium cromoglycate to have a stabilising effect on the mast cell membrane. In practice no new drugs were discovered, since this type of activity in animal models was not predictive of antiasthmatic potential. A more relevant research programme has subsequently evolved, which attempts to approach more closely the conditions prevailing in the asthmatic lung. The use of a model of immune lung inflammation in macaque monkeys in conjunction with a model of bronchial hyper-reactivity in the dog has been successful in producing the new compound, nedocromil sodium, which is proving to be an effective addition to the drugs available for the treatment of inflammatory diseases of the airways. PMID- 2547562 TI - Clinical pharmacology of nedocromil sodium. AB - In attempts to define the clinical pharmacological activity of inhaled nedocromil sodium, various challenge systems, ranging from specific antigen challenge to provocation with chemical irritants such as sulphur dioxide, have been used. A single dose (4 mg) of nedocromil sodium taken before antigen challenge prevented both early and late asthmatic responses, whereas the same dose taken shortly after the early response delayed onset of the late reaction but did not affect its magnitude. Exercise-induced asthma was inhibited by pretreatment with nedocromil sodium, as were the bronchoconstrictor responses to hyperventilation of cold dry air and inhalation of ultrasonically nebulised distilled water ('fog'). Mast cells lying in the surface mucosa of the lung are thought to be less stable in asthmatic subjects and may be implicated in the mechanism of response to these 3 types of physical insult. However, in addition to the marked protective action on isolated mucosal mast cells which has been reported from preclinical studies, nedocromil sodium was also effective against bronchoconstriction induced by sulphur dioxide in hyper-responsive asthmatic and atopic subjects. The response to sulphur dioxide, in which axon reflexes are thought to be involved, is less likely to have an immunological mechanism and it is clear that in this type of situation the effect of nedocromil sodium can be more readily differentiated from that of sodium cromoglycate. The increased potency and wider scope of activity described for nedocromil sodium suggests a therapeutic advantage for this new compound in chronic inflammatory allergic lung disorders. PMID- 2547563 TI - The effect of nedocromil sodium on human eosinophil activation. AB - Nedocromil sodium inhibited the increase in eosinophil activation (measured by IgG- and complement-dependent cytotoxicity assays) induced by platelet activating factor (PAF). Inhibition of the upregulation in eosinophil effector function by nedocromil sodium was dose-dependent (optimal at 10(-7) mol/L) and paralleled that produced by a specific PAF antagonist, BN 52021. In addition, preliminary data suggest that nedocromil sodium can inhibit the increase in IgG-dependent release of LTC4 from human eosinophils after stimulation with the synthetic tripeptide, formyl-methionyl-leucyl-phenylalanine (fMLP). These data support our previous hypothesis that part of the mode of action of nedocromil sodium may be due to its ability to directly block the chemotactic factor-induced enhancement of inflammatory cell activity. PMID- 2547564 TI - Action of nedocromil sodium and sodium cromoglycate on cloned human allergen specific CD4+ T lymphocytes. AB - Nedocromil sodium (and sodium cromoglycate) are topical, prophylactic, anti inflammatory agents used in reversible obstructive airways disease. To investigate their potential mode of action, the effects of these agents on specific allergen- or IL-2-induced proliferation of a human CD4+ house dust mite specific T-cell clone isolated from an individual with perennial rhinitis was examined. Although strong proliferation was observed in control cultures, no inhibitory effect was observed with any of the drug concentrations tested. In contrast, dexamethasone was able to inhibit the T-cell proliferation. These observations suggest that the clinically observed suppression of the late-phase asthmatic response may not be due to an effect on CD4+ T lymphocytes. PMID- 2547565 TI - Effect of nedocromil sodium on the beta-adrenergic response of human polymorphonuclear leucocytes. PMID- 2547566 TI - Nedocromil sodium inhibition of IgE-mediated activation of human mononuclear phagocytes and platelets from asthmatics. AB - The demonstration of a specific receptor for IgE on non-mast cell or basophil leucocytes, such as mononuclear phagocytes, eosinophils and platelets, suggests that these cells may participate directly in immunological disorders of allergy. In this study, inhibition by nedocromil sodium of IgE-dependent activation of human alveolar macrophages, blood monocytes and platelets was investigated. Nedocromil sodium produced an inhibition of IgE-mediated generation of cytotoxic molecules from monocytes and platelets together with a concomitant inhibition of their oxidative metabolism, measured by chemiluminescence. In addition, nedocromil sodium reduced the ability of alveolar macrophages to synthesise and release mediators, estimated by beta-glucuronidase activity. Furthermore, nedocromil sodium inhibited the abnormal response to aspirin of platelets from aspirin-sensitive asthmatics at therapeutic concentrations. These studies confirm that nedocromil sodium acts on a cell compartment other than the classical mast cell population in IgE-dependent allergy and asthma. PMID- 2547567 TI - Effect of sodium cromoglycate and nedocromil sodium on anti-IgE-induced and anti IgG4-induced basophil degranulation. AB - Both anti-IgE and anti-IgG4 induce human basophil degranulation as assessed by toluidine blue staining. Anti-IgG4 has been recently shown to act on the human basophil by a 2-step process: anti-IgG4 induces the release from eosinophils of eosinophil cationic proteins which in turn induce human basophil degranulation. In the present study, we show that sodium cromoglycate and nedocromil sodium have no direct effect on human basophil degranulation but inhibit the anti-IgG4 induced degranulation. This effect was dose-dependent and significant inhibitions were obtained at 2.5 to 25 mumol/L for sodium cromoglycate and 25 mumol/L for nedocromil sodium. No drug effect was observed on the basophil degranulation induced by supernatants from anti-IgG4-stimulated eosinophils. However, the release of basophil degranulating factors (eosinophil cationic proteins) by anti IgG4 from purified eosinophils was significantly inhibited after preincubation with 25 mumol/L of sodium cromoglycate or nedocromil sodium. Taken together, these results indicate that both sodium cromoglycate and nedocromil sodium have an inhibitory effect on the first step of anti-IgG4-induced human basophil degranulation, thus strongly suggesting that these drugs inhibit the release of eosinophil cationic proteins from human eosinophils. PMID- 2547568 TI - Effect of nedocromil sodium on neutrophil and eosinophil-induced epithelial cell desquamation in a human in vitro epithelial model. AB - A human amniotic epithelial membrane preparation was used as a model to study epithelial responses to neutrophils and eosinophils, obtained from normal subjects using Percoll density gradient methods, and activated by phorbyl myristate acetate (PMA). Incubation of activated neutrophils with the epithelial membrane resulted in epithelial cell desquamation, probably due to release of proteases. Activated eosinophils resulted in epithelial cell damage but no desquamation, an action that appeared to be mimicked by major basic protein (MBP). The addition of nedocromil sodium did not significantly inhibit neutrophil induced epithelial cell desquamation. In one preliminary experiment, nedocromil sodium (10(-4) to 10(-5) mol/L) inhibited epithelial cell desquamation induced by a mixed population of eosinophils and neutrophils. PMID- 2547569 TI - Effect of nedocromil sodium on antigen-induced airway responses in allergic sheep. AB - We studied the effects of nedocromil sodium and sodium cromoglycate on early and late bronchial responses, the airway inflammation associated with the late response to inhaled Ascaris suum antigen in allergic sheep in vivo, and the antigen-induced contractile responses of sheep tracheal smooth muscle in vitro. In addition, we examined the effect of nedocromil sodium on the development of antigen-induced airway hyper-responsiveness in this model. Pretreatment with either nedocromil sodium or sodium cromoglycate was effective in blocking antigen induced early and late responses in allergic sheep. Both drugs also prevented the influx of eosinophils into the airways as assessed by bronchoalveolar lavage, observed during the late response in this model. No difference in drug potency was observed in vivo, but in vitro nedocromil sodium was 10-fold more potent than sodium cromoglycate against antigen-induced contractions of sheep tracheal smooth muscle. Nedocromil sodium was effective in blocking antigen-induced late responses and the subsequent development of airway hyper-responsiveness irrespective of whether the drug was given before antigen challenge or after the immediate response to antigen but before the late response. These findings indicate that nedocromil sodium is effective in the sheep model of asthma and therefore may be beneficial in the treatment of reversible obstructive airway disease in man. PMID- 2547572 TI - The pattern of pancreatic carcinoma at Kenyatta National Hospital. AB - A 10-year (1976-1986) retrospective study was done on 30 cases with histological diagnosis of pancreatic carcinoma. The male to female ratio was 1.3:1 and the peak incidence was in the 6th and 7th decades. The head of the pancreas was involved in 96% of cases while solid adenocarcinoma of duct cell origin accounted for 73.3% of cases, followed by anaplastic carcinoma (23.3%). The commonest complications were distinct metastasis (86.6%), obstructive jaundice (73.3%) and upper gastrointestinal bleed (13.6%). PMID- 2547570 TI - Inhibitory effects of nedocromil sodium on the in vitro induced migration and leukotriene formation of human granulocytes. AB - Inflammatory cells, such as neutrophils and eosinophils, are thought to actively contribute to the pathogenesis of asthma since they infiltrate into the lung tissue and may be activated locally to release bronchoconstrictor mediators. In this study we provide evidence that nedocromil sodium is capable of effectively inhibiting the platelet-activating factor (PAF) and zymosan-activated serum (ZAS) induced chemotaxis of polymorphonuclear granulocytes (PMN) [IC50 approximately 1 nmol/L and 0.1 mumol/L respectively]. The same inhibitory potency was obtained with sodium cromoglycate. Thus, nedocromil sodium may effectively inhibit the mobilisation of inflammatory cells in the lung. Furthermore, nedocromil sodium is capable of inhibiting the formation of the bronchoconstrictor mediator leukotriene-C4 (LTC4) by eosinophils in a concentration-dependent way [IC30 for A23187: 5.6 10(-5) mol/L; IC30 for opsonised zymosan (OZ): 6.3 10(-5) mol/L], whereas this drug is not capable of inhibiting leukotriene-B4 (LTB4) formation by neutrophils. These findings indicate that nedocromil sodium inhibits the release of bronchoconstrictor mediators not only from mast cells but also from eosinophils. PMID- 2547571 TI - Effects of nedocromil sodium on airway microvascular leakage and neural reflexes. AB - Microvascular leakage and neurogenic mechanisms may be important components of the inflammatory response in asthmatic airways. We have examined the effects of nedocromil sodium on these responses. In guinea-pig airway, microvascular leakage induced by allergen was significantly inhibited by prior treatment with nedocromil sodium (100 micrograms/kg IV), but this had no effect on histamine induced leakage and reduced PAF-induced leakage only at a dosage of 5 mg/kg. This suggests that nedocromil sodium acts mainly by preventing the release of inflammatory mediators. Both sulphur dioxide (SO2) and bradykinin cause bronchoconstriction in asthmatic patients, which is likely to be due to a neural mechanism since SO2 activates C-fibre sensory nerve endings in airways. Inhaled nedocromil sodium is effective in inhibiting both SO2- and bradykinin-induced bronchoconstriction, and furthermore reduces the sensation of dyspnoea, indicating a possible action on sensory nerve endings in airways. PMID- 2547573 TI - [Carcinogenic activity of synthetic chrysotile asbestos with various fiber sizes and chemical composition]. AB - Synthetic Mg-chrysotiles (analogous to natural ones by chemical composition) with fine short, thick short and fine long fibres under intrapleural injections (20 mg thrice) induced mesotheliomas in noninbred rats in 3.3, 0 and 12% of rats, respectively. Ni- and Co-chrysotiles with fine and short fibres caused such tumours in 7.6 and 30.2%, Co-chrysotile with plate-like fibres induced tumours in 26.8%. The role of chemical structure and particle sizes have been discussed. PMID- 2547574 TI - [The coleukemogenic action of Bordetella pertussis]. AB - Injection of B. pertussis and Rauscher leukemia virus (RLV) in a dose of 4 ID50 to BALB/c mice susceptible to the above virus significantly increases the incidence of leukosis and shortens the average life duration. Injection of B. pertussis to the AKR mice, carriers of the Gross leukosis virus, induces in the first months a greater number of the mice with leukosis and its earlier development. PMID- 2547575 TI - [The activity of growth-regulating substances isolated from the spleen and thymus of healthy cows and of those with hemoblastoses]. AB - Certain agents of the proteoglycan nature possessing organ-specific rather than the species-specific antimitotic chalone activity have been obtained from the normal bovine spleen and thymus. The preparations obtained by the same method from the spleen and the thymus of animals affected by lympholeukemia and lymphosarcoma did not reveal the antimitotic activity; moreover, they were of high mitogenic organ-specific character. A connection between the mitogenic factor produced by lymphocytic cells and the bovine leukemia virus is possible. PMID- 2547576 TI - [Quantifying functional deficits in patients with multiple sclerosis using a computer-assisted visuomotor tracking procedure]. AB - 23 Patients with multiple sclerosis were subjected to a visuomotor tracking test, consisting of subtests of different task difficulty, while being in a state of acute exacerbation and four weeks later, while exhibiting a more or less pronounced clinical remission (Kurtzke DSS 3.9 (+/- 1.4) vs. 3.3 (+/- 1.5); p less than 0.001). Corresponding to this the tracking test performances improved, the difference being significant only with the easiest test run. The tracking performances correlated with the Kurtzke scores, both for the acute state and for the remitted state, the highest correlation showing up for the easiest test run. Hence, the visuomotor tracking test seems to be suited to measure functional deficits and to evaluate the efficiency of therapeutic procedures. PMID- 2547577 TI - [Normative aspects of somatosensory evoked P300 components]. AB - Using a somatosensory version of the oddball-paradigma the influence of age and gender on the P300-component and the comparison of the potential after stimulation of the right and left median nerve was studied in 30 healthy right handed volunteers (age: 20-35 years). Latency, amplitude, area and duration of the P300-potential were analysed. No relationship between age, gender and the P300-parameters were observed. The amplitude and the area of the potential obtained from the F3 electrode were greater after stimulation of the right median nerve compared to the potential after stimulation of the left median nerve. All other results were not significantly different. Strong positive correlations between the results after stimulation of the right and left median nerve were observed. These results showed that by a young group of volunteers age and gender did not influence the P300-component. Although the P300-Parameters had a between subject variability, their mean remained constant over the study, their correlation coefficients were strong positive and the side of stimulation did not influence them (except for the electrode F3). PMID- 2547578 TI - Abnormalities in the hypothalamo-pituitary-adrenal axis in spontaneously hypertensive rats during development of hypertension. AB - Abnormalities in the hypothalamo-pituitary-adrenal axis in spontaneously hypertensive rats (SHR) during development of hypertension were investigated using in vivo and in vitro methods. Plasma ACTH responses to hemorrhage and ether stress were significantly smaller in 7-week-old SHR than in age-matched Wistar Kyoto rats (WKY), while plasma corticosterone baseline levels and its response to stress were greater in SHR than in WKY. There was no significant difference in the plasma ACTH response to ether stress between bilaterally adrenalectomized SHR and WKY replaced with a 25% corticosterone pellet for 6 days. Adrenalectomy prevented the development of hypertension in SHR; however, corticosterone replacement restored hypertension. Plasma ACTH showed a smaller response to iv CRH injection in SHR than in WKY, while the ACTH response to arginine vasopressin was not different between SHR and WKY. CRH concentrations in the median eminence, posterior pituitary, and cerebral cortex were lower in SHR than in WKY, while the CRH concentration in the median eminence was not different in SHR and WKY when they were adrenalectomized with or without corticosterone replacement. Basal in vitro CRH release from hypothalamic tissue was reduced in SHR, while CRH release in response to 56 mM KCl was not different in SHR and WKY. These results suggest that adrenocortical function is enhanced in young SHR, that reduced ACTH response to stress and exogenous CRH in SHR may be ascribed to higher plasma corticosterone levels, and that corticosterone is essential for the development of hypertension in SHR. PMID- 2547579 TI - Effects of tamoxifen, tamoxifen metabolites, and nafoxidine on adenosine 3',5' monophosphate phosphodiesterase: correlations with growth inhibitory activities but not estrogen receptor affinities. AB - Triphenylethylenes [Tamoxifen (TAM), TAM metabolites, and nafoxidine] were found to inhibit Ca2+-calmodulin (CaM)-dependent cAMP phosphodiesterase (PDE) activity of the quail oviduct, whereas 17 beta-estradiol was inactive. The Ca2+-CaM independent PDE activity was not affected by triphenylethylenes, suggesting that they do not interact directly with the active site of the enzyme. Kinetic analysis indicated that these drugs competitively inhibited the activation of PDE by CaM with the following potencies: N-desmethyltamoxifen, Ki = 3 microM; metabolite Z, trans-4-hydroxytamoxifen, and TAM Ki = 5 microM; nafoxidine, Ki = 8.5 microM; and metabolite Y and cis-4-hydroxytamoxifen, Ki = 50 microM. Injected alone into immature quails, none of these drugs significantly affected oviduct weight. When administrated together with estradiol benzoate, these drugs reduced the trophic effect of estradiol in a dose-dependent relationship, with ID50 values ranging from 0.07 mg/kg for N-desmethyltamoxifen to 2.02 mg/kg for cis-4 hydroxytamoxifen. The order of growth inhibitory potency was not correlated with estrogen receptor affinities, but was the same as that reported for PDE inhibition. This correlation suggests that interaction of antiestrogen with Ca2+ CaM dependent PDE may be one of the mechanisms responsible for the estrogen antagonist activity of these drugs. PMID- 2547580 TI - Induction of type II 5'-deiodinase activity in cultured rat astroglial cells by 12-O-tetradecanoylphorbol-13-acetate: dependence on glucocorticoids. AB - The effect of an activator of protein kinase C, 12-O-tetradecanoylphorbol-13 acetate (TPA), on the 5'-deiodinase (5'D) activity was studied in rat astroglial cells cultured in chemically defined medium. TPA promoted a large increase in the type II 5'D activity, which was maximal 5-10 h after addition of TPA and then declined to the basal level at 24 h. The optimal TPA concentration was 10(-7) M. TPA and 8-Br-cAMP, an other inducer of 5'D activity were antagonist. Otherwise, TPA stimulated 5'D activity only in the presence of glucocorticoids at concentrations from 10(-8) M to 10(-4) M. Glucocorticoids alone induced a slight increase in 5'D activity. These data indicate that protein kinase C contributes to the control of 5'D activity in astroglial cells and that its action is dependent on glucocorticoids. PMID- 2547581 TI - Sodium-mediated modulation of aldosterone secretion: impact of converting enzyme inhibition on rat glomerulosa cell response to angiotensin-II. AB - Sodium restriction enhances the aldosterone response to angiotensin-II (AII) in normal rats, but not in spontaneously hypertensive rats (SHR). To determine whether a change and/or abnormality in the circulating or adrenal renin angiotensin systems are responsible for these observations, three groups of animals were studied on a low sodium diet with and without the administration of a converting enzyme inhibitor (enalapril). Sprague-Dawley and Wistar-Kyoto (normotensive rat strains) and SHR were placed on low sodium (0.1%) for 9 days, the last 4 days of which enalapril was administered to half of the animals. In all groups enalapril treatment resulted in a significant (P less than 0.001) reduction in blood pressure, an increase in renin activity, and a reduction in plasma aldosterone when all of the animals were considered together, although the change in blood pressure achieved statistical significance only in the Wistar Kyoto rats. Additionally, basal aldosterone output from isolated glomerulosa cells was lower in the normotensive animals pretreated with enalapril. However, despite the evidence for inhibition of converting enzyme, there was no change in the hypertensive animals. Thus, neither locally nor systemically generated AII appear to participate in the maintenance of the increased aldosterone responsiveness to AII with sodium restriction. Furthermore, they do not appear to contribute to the altered adrenal responsiveness to AII with sodium restriction in SHR. These data provide further support for the hypothesis that as yet undefined glomerulosa intracellular mechanisms are altered by dietary sodium restriction in normotensive, but not hypertensive, rats. PMID- 2547582 TI - Insulin-like growth factor-I potentiates thyrotropin stimulation of adenylyl cyclase in FRTL-5 cells. AB - We reported that TSH and insulin-like growth factor-I (IGF-I), which were known to synergistically stimulate DNA synthesis, synergize to elevate the 1,2 diacylglycerol content of FRTL-5 thyroid cells. We presented evidence that cAMP is the proximal mediator of these actions of TSH. To further define the mechanism of this interaction, we investigated the effects of IGF-I on TSH stimulation of adenylyl cyclase. Long and short term effects of IGF-I or high doses of insulin were studied in FRTL-5 cells that were maintained in serum-, hormone-, and growth factor-free medium for 4-7 days (basal cells). When cells were incubated with high doses of insulin for 7 days and acutely stimulated, a 10-fold increase in sensitivity and a 2-fold increase in maximal responsiveness of cAMP accumulation to TSH were observed. To study shorter term effects, cells were preincubated with insulin for 3 h and then exposed to TSH, cholera toxin, or forskolin. Incubation with high doses of insulin for 3 h caused 30-300% increases in cAMP accumulation at high doses of TSH (greater than or equal to 1 mU/ml), cholera toxin (greater than 0.1 microM), and forskolin, but did not affect the EC50 for TSH. Dose response studies were consistent with insulin acting via receptors for IGF-I, and IGF-I caused a similar effect. There was a 45% increase in adenylyl cyclase activity stimulated by TSH in membranes isolated from cells incubated with high doses of insulin for 3 h. Pretreatment of FRTL-5 cells with pertussis toxin, which ADP-ribosylates the inhibitory G-protein Gi, or adenosine, which we show inhibits cAMP accumulation by interacting with Gi, did not affect insulin/IGF-I enhancement of cAMP accumulation. We suggest that synergism of actions of TSH and IGF-I may in part be due to IGF-I enhancement of TSH stimulation of adenylyl cyclase. PMID- 2547583 TI - Increase in ovarian 15-hydroxyeicosatetraenoic acid during ovulation in the gonadotropin-primed immature rat. AB - The ovarian level of 15-hydroxyeicosatetraenoic acid (15-HETE) was measured by RIA during ovulation in gonadotropin-primed immature Wistar rats. The ovulatory process was initiated in 25-day-old rats by a 10-IU injection of hCG sc 2 days after the animals had been primed with 10 IU PMSG, sc. Ovarian follicles begin to ovulate 10 h after hCG. At 0 h after hCG, the ovarian 15-HETE level was 0.6 +/- 0.2 ng/mg ovarian protein. At 6 h the 15-HETE level increased sharply to 27.3 +/- 4.2 ng/mg protein and reached a peak of 50.0 +/- 9.8 ng/mg protein at 10 h. Ovarian 15-HETE decreased significantly between 10-16 h after hCG (when ovulation was essentially completed). The pattern of secretion of this 15-lipoxygenase product was reciprocal to the pattern of secretion of leukotriene-B4 by the rat ovary. Ovarian 15-HETE production and ovulation were inhibited in a dose dependent manner when indomethacin was administered sc 1 h after hCG in doses ranging from 0.10-10.0 mg/rat. In contrast, the synthesis of ovarian prostaglandins-E and -F was inhibited by a dose of indomethacin as low as 0.0316 mg/rat, but this dose did not significantly affect the ovarian 15-HETE level or the ovulation rate. Therefore, the ovulation rate was more closely correlated with the ovarian 15-HETE level (P less than 0.001) than with the ovarian levels of either prostaglandin-E or -F (0.10 greater than P greater than 0.05). The results suggest that products of the 15-lipoxygenase pathway of arachidonic acid metabolism may be important in the biochemical events of mammalian ovulation. PMID- 2547584 TI - Effects of prolonged cyclosporine-A treatment on the morphology and function of rat adrenal cortex. AB - The effects of prolonged (30 day) treatment with daily therapeutical doses of cyclosporine A (CSA) (20 mg/kg) on the function and morphology of adrenal cortex were studied in adult male rats. CSA-treated animals developed a notable hypertension, along with a striking rise in PRA, which was not coupled with significant changes in the plasma concentrations of aldosterone and corticosterone (hyperreninemic hypoaldosteronism). Morphometry showed that zona glomerulosa (ZG) and zona fasciculata, and their parenchymal cells were atrophic. Isolated capsular (ZG) and inner (zona fasciculata/reticularis) cells displayed reduced basal and stimulated secretory responses. However, while the response of ZG cells to angiotensin II was almost completely suppressed (96%), basal steroid secretion of isolated cells, as well as the aldosterone and corticosterone response of ZG cells to potassium and ACTH, and corticosterone production of inner cells in response to ACTH were decreased by only about 30-40%. The hypothesis is advanced that CSA exerts a dual effect on rat adrenal cortex: 1) a general inhibitory effect on the growth and steroidogenic capacity of adrenocortical cells, which manifests itself only after very prolonged treatment and may be caused by an impairment of protein synthesis; and 2) an acute effect involving the specific blockade of the angiotensin-II-induced stimulation of the secretory activity of ZG cells. PMID- 2547585 TI - Time course of the estradiol-dependent induction of oxytocin receptor binding in the ventromedial hypothalamic nucleus of the rat. AB - Oxytocin (OT) transmission is involved in the steroid-dependent display of sexual receptivity in rats. One of the biochemical processes stimulated by the ovarian steroid 17 beta-estradiol (E2) that is relevant to reproduction is the induction of OT receptor binding in the ventromedial hypothalamic nucleus (VMN). The purpose of these experiments was to determine if E2-induced changes in OT receptor binding in the VMN occur within a time frame relevant to cyclic changes in ovarian steroid secretion. OT receptor binding was measured in the VMN of ovariectomized rats implanted for 0-96 h with E2-containing Silastic capsules. The rate of decay of OT receptor binding was measured in another group of animals 6-48 h after capsule removal. Receptors were labeled with the specific OT receptor antagonist [125I]d(CH2)5[Tyr(Me)2,Thr4,Tyr-NH2(9)]OVT, and binding was measured with quantitative autoradiographic methods. In addition, plasma E2 levels and uterine weights were assessed in animals from each treatment condition. Significant increases in E2-dependent OT receptor binding and uterine weight occurred within 24 h of steroid treatment. After E2 withdrawal, OT receptor binding and uterine weight decreased significantly within 24 h. These results are consistent with the hypothesis that steroid modulation of OT receptor binding is necessary for the induction of sexual receptivity. PMID- 2547586 TI - Insulin-stimulated phosphorylation of a 195K protein from muscle and liver in the presence of poly-L-lysine. AB - Substrates of the insulin receptor tyrosine kinase have not been identified in skeletal muscle, a major target organ of insulin action. We observed the insulin stimulated phosphorylation of a 195K protein (pp195) in extracts prepared from rat skeletal muscle and liver. pp195 copurifies with the insulin receptor on wheat germ agglutinin affinity chromatography. pp195 is not related to the insulin receptor, as assessed by lack of recognition by antinsulin receptor antibodies and by phosphopeptide mapping. Reduction of sulfhydryl bonds does not affect its apparent mol wt. Phosphorylation of pp195 has an absolute requirement in vitro for Mn2+ or Mg2+ and for certain basic poly-amino acids, i.e. poly-L lysine or poly-L-ornithine. In the presence of 1 microM poly-L-lysine insulin stimulates pp195 phosphorylation in a dose-dependent manner (k0.5, approximately 5 x 10(-10) M; maximum approximately 10(-8) M insulin); pp195 phosphorylation by insulin-like growth factor-I requires about 100-fold higher doses. By phosphoamino acid analysis, pp195 is predominantly phosphorylated on tyrosine, and it is recognized by antiphosphotyrosine antibodies. Insulin receptors isolated from rat muscles 5 min after insulin injection induce about 2-fold greater phosphorylation of pp195 in vitro than receptors isolated from saline injected controls. Streptozotocin-induced diabetes results in marked diminution of insulin-stimulated pp195 phosphorylation in extracts of muscle and liver (approximately 50% when normalized to protein content of wheat germ agglutinin eluates or approximately 80% reduction when normalized to equal receptor number). The defect is reversible by insulin therapy in vivo. PMID- 2547587 TI - Evidence that luteinizing hormone-stimulated differentiation of purified ovarian thecal-interstitial cells is mediated by both type I and type II adenosine 3',5' monophosphate-dependent protein kinases. AB - LH has been shown to be the principal hormone regulating ovarian thecal interstitial cell (TIC) differentiation. It has been well documented that LH stimulates cAMP production and that cAMP analogs mimick the stimulatory actions of LH, but the mechanisms by which LH and cAMP stimulate TIC differentiation are unknown. The purpose of these studies was to characterize LH-stimulated differentiation of isolated TIC in serum-free medium and examine the role of cAMP dependent protein kinase (PKA) isoenzymes in TIC differentiation. Highly purified (greater than 90%) TIC which were free from granulosa cell contamination were isolated from collagenase-dispersed ovaries of hypophysectomized immature rats by Percoll gradient centrifugation. When the purified TIC (20,000 viable cells/well) were cultured (2 days) in serum-free medium (0.2 ml in 96-well plates), low levels of steroids were produced. LH stimulated a dose-related (ED50 = 2.6 +/- 0.4 ng/ml) increase (50-fold) in androsterone, the principal androgen produced. LH stimulated an immediate dose-related increase in cAMP production, but there was a 20-h lag before LH stimulated an increase in androsterone production, which reached maximum levels at 30 h. LH-stimulated progesterone production increased immediately to a maximum at 10 h, then progesterone levels decreased as androsterone production increased. To determine the role of PKA in stimulating androsterone and progesterone production, TIC were cultured (2 days) with 8 aminohexylamino-cAMP (100 microM) plus N6-benzoyl-cAMP (30 microM) or 8 thiomethyl-cAMP (30 microM) plus N6-benzoyl-cAMP (30 microM) to directly and selectively activate type I or type II PKA, respectively. Selective activation of either isoenzyme increased androsterone and progesterone production by TIC. Immunoblots revealed that either type I or type II PKA increased the contents of P450scc and P45017 alpha in TIC. This is the first demonstration that direct activation of either type I or type II PKA stimulates TIC differentiation. These results indicate that LH stimulates TIC differentiation by a mechanism mediated by activation of one or both PKA isoenzymes. PMID- 2547588 TI - Nutritional regulation of the placental lactogen receptor in fetal liver: implications for fetal metabolism and growth. AB - We have recently identified and purified from fetal liver a distinct receptor that mediates the effects of placental lactogen (PL) on amino acid transport, glycogen synthesis, and somatomedin production in fetal tissues. At present, the factors that regulate the number and affinity of PL receptors in the fetus are unknown. Since maternal nutrition plays a critical role in fetal metabolism and growth, we have examined the role of nutrition in the regulation of the PL receptor in fetal lambs. Pregnant ewes at 123-126 days gestation were fed ad libitum (FED), fasted for 3 days (FASTED), or fasted for 3 days and then refed for an additional 3 days (REFED). The ewes were then killed, and the binding of [125I]ovine (o) PL to hepatic microsomes from the fetal lambs was examined. Maternal fasting caused a 60-75% reduction in the specific binding of oPL to fetal liver; the effect of fasting was reversed in part by refeeding [specific binding per mg protein: FED, 11.8 +/- 2.2% (n = 8); FASTED, 2.8 +/- 0.4% (n = 7); REFED, 7.2 +/- 2.6% (n = 3)]. The decrease in oPL binding resulted from an 80% reduction in the number of fetal oPL-binding sites (Scatchard analysis); there were no changes in the affinity of the oPL receptor (Kd, 0.6 nM), the subunit structure of the receptor, or the degree of occupancy of the receptor in vivo by endogenous fetal hormones. The specific bindings of GH (0.6%), PRL (0.3%), and insulin (35%) to fetal liver were not affected by maternal fasting, indicating that caloric restriction exerted a specific effect on oPL binding in the fetus. The number of fetal oPL-binding sites was positively correlated with the fetal liver glycogen content (r = 0.69; P less than 0.01) and the fetal plasma concentrations of glucose (r = 0.68; P less than 0.01) and insulin-like growth factor-I (r = 0.74; P less than 0.001), suggesting a role for the PL receptor in the regulation of fetal carbohydrate metabolism and growth. The number of fetal PL receptors was inversely correlated with the fetal plasma oPL concentration (r = 0.47; P less than 0.05). These studies demonstrate that fasting of the pregnant ewe reduces the number of PL receptors in ovine fetal liver. The reduction in fetal hepatic PL receptors may contribute to the mobilization and depletion of fetal liver glycogen stores and may play a role in the pathogenesis of the fetal growth retardation that accompanies maternal caloric deprivation. PMID- 2547589 TI - Effect of estrogen on pituitary peptide-induced dehydroepiandrosterone secretion in the baboon fetus at midgestation. AB - We have previously shown that ACTH and PRL stimulate baboon fetal adrenal dehydroepiandrosterone (DHA) production both in vitro and in vivo and that estrogen diminishes the responsivity of the adrenal to tropic peptides in vitro. In the present study we determined the effects of increasing placental estrogen production by the administration of androstenedione at midgestation on DHA production by the baboon fetus in vivo. Pregnant baboons were untreated (n = 8) or treated (n = 9) with increasing numbers of androstenedione implants inserted in the mother at 8-day intervals between days 70-100 of gestation (term = day 184). On day 100, the fetuses were exteriorized, and a constant infusion of saline (0.1 ml/min) was initiated via a catheter inserted into a femoral vein of the fetus. At 40 min, a bolus injection of either 30 nmol ACTH or 40 nmol ovine PRL was administered to fetuses. ACTH or PRL (0.2 nmol/min.0.1 ml saline) were then infused for an additional 25 min. The concentrations of serum estradiol (E2) in the uterine vein (20.2 +/- 1.5 ng/ml; mean +/- SE) and estrone (E1) in umbilical vein (11.9 +/- 3.1 ng/ml) of androstenedione-treated baboons were 2 fold greater (P less than 0.05) than respective values in untreated baboons. Baseline concentrations of DHA in the femoral vein of the fetus were similar in all treatment groups (overall mean, 120 +/- 20 ng/ml) and greater (P less than 0.05) than values (27 +/- 3) in the mother. In untreated control baboons, basal DHA concentrations in the fetus were increased (P less than 0.05) by 69 +/- 17% and 94 +/- 29% after fetal injection of ACTH (n = 4) or PRL (n = 4), respectively. In contrast, neither PRL (n = 5) nor ACTH (n = 4) had any effect on serum DHA when injected into androstenedione-treated baboons. Regardless of treatment, injection of ACTH or PRL into the fetus had no effect on DHA concentrations in the mother. Collectively, these findings indicate that the ability of the fetal adrenal to increase DHA production in response to an acute infusion of ACTH or PRL was abolished in baboons in which placental estrogen production was increased prematurely at midgestation. Therefore, we suggest that during the second half of gestation in the baboon a regulatory system may exist in utero, in which there is feedback control of the placental product estrogen on the formation of the fetal adrenal precursor DHA. PMID- 2547590 TI - Structure, specificity, and regulation of the insulin-like growth factor-binding proteins in adult rat serum. AB - Insulin-like growth factor-I (IGF-I), the principal IGF in adult rat serum, occurs complexed to specific binding proteins. After fractionation of serum on Sephadex G-200 at neutral pH, 62% of the immunoreactive IGF-I is recovered in the 150K region, 38% in the 40K region, and none is present as free 7.5K IGF-I. Adult rat serum also contains unoccupied binding sites for IGFs that also are predominantly (77%) located in the 150K region and have preferential binding affinity for IGF-II. IGF-binding protein components in the 150K and 40K regions were evaluated by affinity cross-linking to 125I-labeled IGFs and by ligand blotting (i.e. incubation of nitrocellulose blots of sodium dodecyl sulfate (SDS) gels with [125I]IGFs). Affinity cross-linking of the 150K region revealed a major 43K binding protein complex and several minor covalent complexes of 97-210K that are formed during the cross-linking reaction. The 40K region of the gel filtration column contains a predominant 32K complex and smaller amounts of the 43K complex. Ligand blotting of the 150K region identifies a predominant cluster of binding components of about 40K and a smaller 29K protein. The apparent molecular masses of the 40K and 29K proteins are decreased by incubation with N glycanase, indicating that they contain N-linked oligosaccharides. These glycoprotein components, designated gp40 and gp29, presumably combine with an acid-labile nonbinding subunit of about 100K to generate the 150K complex. The gp40 cluster represents glycosylation variants of a 34K protein; gp29 has been shown to correspond to an amino-terminal fragment of gp40. Ligand blotting of the 40K region indicates that it contains smaller amounts of gp40 and gp29, possibly representing free subunits not combined with the nonbinding subunit, as well as two proteins of apparent molecular mass 24K and 30K (p24 and p30) that are not glycosylated. Although p30 is similar in size to the binding protein from BRL-3A cells (BP-3A) that is present in fetal rat serum, immunoprecipitation and immunoblotting of whole and fractionated adult serum with an antiserum to BP-3A indicate that p30 in adult rat serum is an antigenically distinct protein. Serum levels of gp40 and gp29 are decreased by hypophysectomy and are restored by GH treatment; p24 and p30 show similar but smaller changes. PMID- 2547591 TI - Calcitonin receptors as markers for osteoclastic differentiation: correlation between generation of bone-resorptive cells and cells that express calcitonin receptors in mouse bone marrow cultures. AB - The osteoclast is the cell that resorbs bone. It is known to derive from hemopoietic precursors, but analysis of lineage and regulation of differentiation has been hampered by lack of a specific marker that enables identification of cells of osteoclastic phenotype. Previously used markers, such as multinuclearity, that are specific for osteoclasts in bone become less specific in culture. Uniquely among bone and bone marrow cells, osteoclasts possess abundant calcitonin (CT) receptors. We therefore tested the correlation between the generation of bone-resorptive function and the formation of CT receptor positive cells from hemopoietic tissue in vitro. Without 1,25-dihydroxy-vitamin D3 [1,25-(OH)2D3], a hormone that induces osteoclastic differentiation in vitro, bone marrow cultures showed very little bone resorption, and only small numbers of CT receptor-positive cells developed. When 1,25-(OH)2D3 was added to the cultures, CT receptor-positive cells developed within 1 day and reached a peak after 7 days. Bone resorption commenced within 2 days of hormone addition. There was a strong parallelism between the cumulative number of CT receptor-positive cells and the extent of bone resorption. The capacity of cultures to generate bone-resorptive activity and CT receptor-positive cells declined progressively when 1,25-(OH)2D3 was added to hemopoietic tissue after a 7- to 21-day hormone free incubation period. The number of CT receptor-positive cells in these cultures correlated strongly (r = 0.96) with bone resorption. The behavior of these cultures suggests that 1,25-(OH)2D3 acts to induce terminal differentiation of osteoclast precursors present in the cultures, and that precursor cell numbers decreased with increasing time in vitro. All of the CT receptor-positive cells in control cultures and all of those seen shortly after 1,25-(OH)2D3 addition were mononuclear, despite considerable bone resorption; the majority of CT receptor positive cells remained mononuclear throughout the incubation period. This suggests that mononuclear cells with characteristics of osteoclasts exist that are able to excavate bone. CT receptor-positive cells slightly preceded the development of bone-resorptive function, implying that CT receptors develop before the acquisition of bone-resorptive capacity by osteoclasts. Peritoneal macrophages, blood mononuclear cells, and cells of the J774 macrophage cell line failed to either resorb bone or express CT receptors, even after incubation with 1,25-(OH)2D3 for 14 days. These results show a strong and specific correlation between the generation of bone-resorptive cells and CT receptor-positive cells, and suggest that CT receptor express PMID- 2547592 TI - Synthesis of gonadotropin-releasing hormone receptors by gonadotrope cell cultures: both preexisting receptors and those unmasked by protein kinase-C activators show a similar synthetic rate. AB - Responsiveness of gonadotropes to GnRH depends, in part, on the number of plasma membrane GnRH receptors. Since the steady state level of these plasma membrane receptors is a function of the rates of both receptor generation (synthesis, unmasking, and recycling) and loss (internalization, degradation, and inactivation) we have sought to quantify the rate of synthesis of GnRH receptors in pituitary cell cultures. Further, since the protein kinase-C activator phorbol 12-myristate 13-acetate (PMA) has been shown to unmask a class of GnRH receptors that appear to be uncoupled from phosphoinositide turnover, we have measured the rate of synthesis of this second receptor population. The present studies use the density shift technique; incorporation of densely labeled amino acids confers a higher density to newly synthesized proteins and allows their separation by physical means. Cultures of pituitary cells were prepared from female weanling rats. After cells had attached to the culture dishes, medium was replaced at 12-h intervals with medium containing either densely labeled or normal amino acids. After the incubation, GnRH receptors were covalently linked to a photoaffinity receptor agonist [( 125I]Tyr5-[azido-benzoyl-D-Lys6-GnRH]), then solubilized with 1+ sodium dodecyl sulfate. In some cultures PMA (50 nM) was included during the photoaffinity agonist-binding step. Newly synthesized (dense) receptors were separated from previously synthesized receptors by velocity sedimentation (0-20% sucrose in 1% sodium dodecyl sulfate-10 mM Tris-HCl, pH 7.0; centrifuged at 156,000 x g for 24 h). Gradients were fractionated, and the radioactivity contained in each fraction was quantified. Newly synthesized GnRH receptors exhibited a higher density, as evidenced by further migration into the gradient, than did normal GnRH receptors. There was a delay of approximately 6 h between exposure to dense amino acids and the appearance of densely labeled GnRH receptors at the plasma membrane. Equilibrium for incorporation of dense amino acids into GnRH receptors was 48 h of exposure to dense amino acids. The time required for synthesis of half the entire population of GnRH receptors was 28 +/- 2 h (mean +/- SEM; n = 4). Scatchard analysis and the pattern of GnRH-stimulated LH release from densely labeled cells indicated that they bound the photoaffinity label (Kd = 0.4 nM; approximately 1 fmol receptor/microgram DNA) and secreted gonadotropin normally. Additionally, treatment with PMA caused a significant increase (181 +/- 24%) in photoaffinity agonist binding, consistent with previous observations. PMID- 2547593 TI - The intrarenal localization of mineralocorticoid receptors and 11 beta dehydrogenase: immunocytochemical studies. AB - The 11 beta-dehydrogenase (11-DH) component of the microsomal enzyme complex, 11 beta-hydroxysteroid dehydrogenase, has been postulated to be the specificity conferring mechanism for the renal type I receptor. To this end, the distribution of 11-DH and the type I receptor was studied in rat kidney by immunocytochemistry. 11-DH-like immunostaining (LI) was found in Bouin's fixed tissue, in both nuclear and cytoplasmic components of some cells of the juxtamedullary proximal convoluted tubules, and in interstitial cells of the papillary medulla. In addition, weak staining was seen in superficial proximal convoluted tubules in paraformaldehyde-fixed tissue. Type I receptor-LI was found in distal convoluted tubules, connecting pieces, and initial cortical collecting tubules of the superficial cortex. Since colocalization of the enzyme and receptor was not demonstrated, the action of 11-DH on type I receptor specificity would, thus, appear to be paracrine rather than autocrine. PMID- 2547594 TI - GnRH receptors in human granulosa cells: anatomical localization and characterization by autoradiographic study. AB - The presence of receptors for GnRH in human ovary has been investigated by quantitative autoradiography. Simultaneous visualization and characterization of specific receptors on frozen sections were obtained on six pairs of human ovaries. Among them only one exhibited a large preovulatory follicle. This dominant follicle exhibited a specific and high affinity binding capacity for 125I-GnRHa exclusively localized on the granulosa cell layer. Analysis of saturation curve indicates a Kd value of 0.22 nM and Bmax of 9.6 fmol/mg protein. In contrast LH-hCG binding sites were present in all antral follicles. These data demonstrate for the first time the presence of high affinity GnRH receptors in human granulosa cells at a late stage of follicular maturation. PMID- 2547595 TI - Subcellular localization of a protein produced in adrenal cortex cells in response to ACTH. AB - We have reported previously that a protein, ib, is produced in adrenal cortex and other steroidogenic cells with the same tissue-specific peptide hormone or cAMP dose-response and the same kinetics as the increase in steroid hormone biosynthesis. In this study, we have fractionated adrenal cortex cells into subcellular components and used two-dimensional electrophoresis to characterize the proteins in these fractions. We have demonstrated previously that inhibition of cytosolic translation, e.g. by cycloheximide, prevents the production of protein ib. We also report that the production of this protein is not affected by inhibition of mitochondrial translation by chloramphenicol. PMID- 2547596 TI - Isolation and biological activity of corticostatic peptides (anti-ACTH). AB - Corticostatins (CS's) are a family of low molecular weight peptides, rich in arginine and cysteine with the ability to inhibit ACTH stimulated adrenocortical steroidogenesis. They show a high degree of specificity in that they do not inhibit the action of angiotensin II in the adrenal cortex. Four corticostatins have been isolated from rabbit lung extracts and peritoneal neutrophil extracts and one from human neutrophils. Among them corticostatin I, CSI, is the most potent with an I.D.50 of 25 nM. Corticostatin activity is different from other published inhibitory factors such as TGF-B and ANF, which inhibit basal and angiotensin II stimulated steroidogenesis. Other factors such as macrophage secreted products, and septic shock plasma factors which have not been fully characterized also have suppressive activity on ACTH induced adrenocortical steroidogenesis in vitro. It is not yet clear whether there is any relationship between corticostatins and these macrophage products and shock plasma factor. Corticostatins do not inhibit dbcAMP induced steroidogenesis, however they do inhibit the accumulation of cAMP in response to ACTH in rat adrenal cell suspensions. Binding studies show that CSI is a competitive inhibitor of ACTH, probably acting by blocking, the address recognition site of the receptor. There is wide variation in the potency of the corticostatins ranging from an ID50 of 100 ng/ml for CSI to a completely inactive analog, HP1. In this paper we will describe the purification of the corticostatins and some recent results obtained on their mechanism of action. PMID- 2547597 TI - Regulation of steroid 17 alpha-hydroxylase in adrenocortical cells in culture. AB - The regulation of steroid 17 alpha-hydroxylase in human and bovine adrenocortical cells in culture is reviewed. It is shown that (i) the long-term growth and cloning of normal human fetal adrenocortical cells in culture is feasible; (ii) the phorbol ester 12-O-tetradecanoyl phorbol 13-acetate (TPA) is an effective mitogen in both clonal cultures and non-clonal early cultures of human adrenocortical cells, and shows a selective action in promoting adrenocortical cell growth and inhibiting fibroblast growth; (iii) the key steroidogenic enzymes, 17 alpha-hydroxylase and 3 beta-hydroxysteroid dehydrogenase (3 beta HSD), are under dual regulation by the cyclic AMP and protein kinase C second messenger systems; (iv) for 17 alpha-hydroxylase, this dual regulation is mediated by changes in 17 alpha-hydroxylase mRNA levels; (v) bovine adrenocortical cells can be transfected with SV40 T antigen, producing lines with elevated differentiated functions, including stabilized high expression of 17 alpha-hydroxylase; (vi) human adrenocortical cells can also be transfected with SV40 large T antigen, giving rise to functional cell lines which may be useful in future studies of 17 alpha-hydroxylase regulation. PMID- 2547598 TI - Regulation of 21-hydroxylase gene expression. AB - The steroid 21-hydroxylase (21-OH) gene is selectively expressed in the adrenal cortex and is transcriptionally regulated by ACTH. We examined the role of the 5' flanking sequences of 21-OH in this regulated expression by analyzing their ability to direct the expression of a human growth hormone (hGH) reporter gene upon transfection into Y1 mouse adrenocortical tumor cells. The 330 bp of 5' flanking sequences directed basal and hormonally-inducible expression of hGH in Y1 cells, but did not direct expression in I-10 mouse testicular Leydig cells. Both constitutive and hormonally-inducible expression required a functional cAMP dependent protein kinase. These results indicate that the first 330 bp of 5' flanking sequences of the 21-OH gene contain sufficient information for cell specific and hormonally regulated expression, and that this expression requires the integrity of cAMP-dependent protein kinase. Markedly lower expression of hGH was seen when 156 bp of 5'-flanking sequences were placed in front of the reporter gene, suggesting that sequences between -330 and -156 are essential for expression. The addition of sequences from -330 to -150 to the p-156GH plasmid, in either the correct or the reverse orientation, restored promoter activity to approximately the level obtained with the 330 bp of 5'-flanking sequences. Moreover, the addition of sequences from -230 to -150 increased by 5-fold the expression of hGH driven by the heterologous thymidine kinase promoter. Based on these results, we conclude that an enhancer element is contained within the sequences from 230 to 150 bp upstream of the transcription initiation site. PMID- 2547599 TI - Recovery of responsiveness to ACTH and cAMP in a protein kinase-defective adrenal cell mutant following transfection with a protein kinase gene. AB - The cause and effect relationship between mutations in cAMP-dependent protein kinase activity and resistance of adrenocortical tumor cells to ACTH and cAMP was evaluated by transfection with cloned cDNAs encoding subunits of the mouse cAMP dependent protein kinase. Protein kinase defective, Kin 8 adrenocortical tumor cells were transfected with pRev [an expression vector encoding the regulatory subunit of the type 1 cAMP-dependent protein kinase (RI)] or with pC alpha ev [an expression vector encoding the catalytic subunit of cAMP-dependent protein kinase (C)]. The pC alpha ev transformant recovered cAMP responsive protein kinase activity, whereas the pRev transformant recovered cAMP-binding activity, but did not recover cAMP responsive protein kinase activity. The pC alpha ev transformant concomitantly recovered steroidogenic and morphologic responsiveness to ACTH- and 8-bromo-cAMP, whereas the pRev transformant remained resistant to these effects of the hormone and cyclic nucleotide. Since Kin 8 cells recovered their responsiveness to ACTH and 8-bromo-cAMP following transfection with pC alpha ev we suggest that the defect in cAMP-dependent protein kinase activity is directly responsible for the ACTH- and cAMP-resistant phenotype of the Kin 8 mutant. PMID- 2547600 TI - Sensitization of human fibroblasts to insulin-like growth factor I by serum deprivation and dexamethasone pretreatment. AB - Pretreatment of confluent human fibroblast cultures for two days in dexamethasone, serum free medium increased 10-20 fold the sensitivity of the cells to insulin-like growth factor I (IGF I) stimulation of amino acid uptake using the amino acid analog alpha-aminoisobutyric acid (AIB). This increased sensitivity resulted both from the use of serum free medium and the addition of dexamethasone to the serum free media. Pretreatment of the cells for 1, 2, or 3 days before assay showed that the maximum increase in sensitivity was obtained after a two day pretreatment. Pretreatment of the cells also increased their sensitivity to insulin and bovine insulin-like growth factor II stimulation of AIB uptake similar to that seen for IGF I. No consistent effect of the pretreatment was observed on either the basal level of AIB uptake or the maximal hormonal stimulation of AIB uptake. Nor was any change noted in the shape of the dose response curves. Addition of IGF I to the pretreatment medium greatly reduced the sensitivity of pretreated cells. [125I]IGF I binding studies done on suspended fibroblasts indicated that there was up to a two fold increase in the number of receptors with no increase in their affinity for IGF I. Thus, pretreatment of fibroblasts with dexamethasone and serum free medium greatly enhances their sensitivity to IGF I stimulation of AIB uptake and makes this an excellent in vitro bioassay system for IGF I. PMID- 2547601 TI - The role of calcium in gonadotropin-releasing hormone induction of follicle stimulating hormone release by the pituitary gonadotrope. AB - Binding of gonadotropin-releasing hormone (GnRH) to the pituitary gonadotrope induces activation of a membrane associated calcium channel, resulting ultimately in luteinizing hormone release. The role of calcium mobilization in GnRH-induced follicle-stimulating hormone (FSH) release was explored using anterior pituitary glands from female rats in a perifusion tissue culture system. While perifusion with GnRH (10 ng/ml) induced a constant level of gonadotropin release, the calcium channel blocker verapamil (10(-4)M) depressed FSH release, as did dantrolene (10(-4)M), an antagonist of intracellular calcium mobilization. When the calcium ionophore A23187 (10(-5) M) was substituted for GnRH, FSH release was not only maintained but increased. Antagonism of the activity of calmodulin (CAM) with trifluoperazine (10(-4)M), however, did not depress FSH release. Cellular content of cAMP and cGMP increased in response to GnRH. When FSH secretion was ionophoretically induced by A23187, however, little cAMP was detected. These results support a role for calcium mobilization in the second messenger cascade underlying GnRH-induced FSH release. The role for calcium in the disparate release of FSH and LH were further discussed in the context of these data. PMID- 2547602 TI - The insulin-like effects of phorbol myristate acetate (PMA) in the isolated fat cell. AB - Recent data from many laboratories suggest that insulin stimulates diacylglycerol formation. Data presented in this manuscript demonstrate an insulin-like effect of PMA, a tumor promoting agent that mimics the action of diacylglycerol, in isolated adipocytes on; (a) glucose oxidation using uniformly labelled, C-1 labelled and C-6-labelled glucose, (b) epinephrine-induced lipolysis and (c) low Km cAMP phosphodiesterase activity. Additionally, a lipolytic effect of PMA is identified when unopposed by epinephrine. These data not only demonstrate an insulin-like effect of phorbol esters in adipose tissue but they lend support to the concept of diacylglycerol involvement in the mechanism of insulin action. PMID- 2547603 TI - Leukotrienes and the airways. PMID- 2547604 TI - Mitogen-responsive S6 kinase. AB - Many cell lines respond to mitogenic stimuli (serum, growth factors) with rapid phosphorylation of the ribosomal protein S6 at several serine sites. We have tried to identify the protein kinase(s) mediating this effect of growth stimuli. Examining post-DEAE chromatography fractions of S49 kin- cell extracts, we could detect a highly active effector-independent S6 kinase with specificity for serine residues. The study was extended to the presumably homologous human enzyme, using HeLa S3 cells as model system. Activity yields increased up to sevenfold when exhausted HeLa cells were supplied with fresh medium plus serum. The enzyme uses ATP, not GTP, as cosubstrate, 40-S or 80-S (reassociated from subunits) ribosomal particles being substrate. The optimal K+ concentration, measured at 3 mM Mg2+, is 35 mM. Under optimized assay conditions S6 phosphorylation proceeded faster in vitro than it appeared to do in vivo. The apparent Mr of the enzyme, as estimated by gel filtration on Sephadex G-100, is 56,000 (determination in the presence of 200 mM KCl in 25 mM phosphate buffer). Tighter binding to DEAE-Sephacel and higher specificity for S6 distinguishes this enzyme from the following S6 phosphorylating protein kinases: protein kinase C, protease-activated kinase II, histone-4 phosphotransferase and an enzyme with the properties of casein kinase I. In published summaries of observations shown here and in a follow-up study with chick embryo fibroblasts, the enzyme(s) has been referred to as mitogen responsive S6 kinase(s) [Martini, O. H. W. and Lawen, A. (1985) in Hormones and cell regulation (Dumont, J. E., Hamprecht, B. and Nunez, J., eds) vol. 9, pp. 411 412, Elsevier Company, North-Holland, Amsterdam; Lawen, A. and Martini, O. H. W. (1985) FEBS Lett. 185, 272-276]. PMID- 2547605 TI - Insulin-induced S6 kinase activation in HeLa cells and its reversal by hyperthermic stress. AB - Insulin treatment of HeLa S3 cells activates an S6-phosphorylating protein kinase. Although this enzyme has chromatographic properties resembling those of described proteolytic fragments of other protein kinases, namely protein kinase C, protease-activated kinase II and histone-4 protein kinase, and although insulin has been proposed by others to cause S6 phosphorylation via proteolytic protein kinase activation, the insulin-induced increase in S6-kinase activity described here is probably not due to proteolysis. Rather, the activity indicates the existence, in HeLa cells, of an interconvertible S6 kinase, since the insulin induced activity increase was rapidly reversed under hyperthermic stress, and since this effect of hyperthermia was itself reversible. The S6-kinase activities from serum- and from insulin-stimulated HeLa cells resemble each other closely and are likely to represent the same enzyme. The enzyme may therefore mediate both signals delivered by mitogens and the insulin signal. Analysed at an in vitro transfer of 1 mol phosphate/mol S6, this S6 kinase activity does not phosphorylate the (principal) S6 site recognized by the cAMP-dependent protein kinase. PMID- 2547606 TI - Properties of vasoactive-intestinal-peptide receptors and beta-adrenoceptors in the murine radiation leukemia-virus-induced lymphoma cell line BL/VL3. AB - 1. Based on radioligand binding and adenylate cyclase activation, functional receptors to vasoactive intestinal peptide(VIP)/helodermin, were shown to coexist with beta 2-adrenoceptors and prostaglandin receptors in membranes from a cultured cloned BL/VL3 cell line of murine T-cell lymphoma induced by a radiation leukemia virus. 2. The relative potency of VIP-related peptides to stimulate adenylate cyclase activity was: helodermin greater than VIP greater than peptide histidine isoleucinamide. Five VIP analogs inhibited 125I-iodo-VIP binding and stimulated adenylate cyclase activity, their decreasing order of potency being: VIP greater than [D-Asp3]VIP greater than [D-Ser2]VIP greater than [D-Ala4]VIP = [D-His1]VIP = [D-Phe2]VIP. [D-Phe2]VIP acted as a partial agonist (with an intrinsic activity of 0.1 as compared to that of VIP = 1.0) and competitively inhibited helodermin- and VIP-stimulated adenylate cyclase activity with a similar Ki (0.07-0.10 microM). These data suggest the existence, in this murine T cell lymphoma, of VIP receptors of the 'helodermin-preferring' subtype that are coupled to adenylate cyclase. PMID- 2547607 TI - Homologous and heterologous regulation of the helodermin/vasoactive-intestinal peptide response in the murine radiation leukemia-virus-induced lymphoma cell line BL/VL3. AB - 1. Functional vasoactive intestinal peptide (VIP)/helodermin receptors and beta 2 adrenoceptors coexist in membranes from a cultured cloned BL/VL3 cell line of murine T-cell lymphoma induced by a radiation leukemia virus (see preceding paper in this journal). 2. Short-term (5-30 min) exposures of BL/VL3 cells to VIP or isoproterenol induced both homologous and heterologous desensitization. The potency of VIP and isoproterenol to desensitize was similar to their potency to occupy receptors and activate adenylate cyclase. 3. Long-term (16-h) exposure of BL/VL3 cells to VIP induced homologous down regulation only, whereas isoproterenol induced both homologous and heterologous down regulation. The potency of VIP, peptide histidine isoleucinamide, helodermin, helospectin, and [D Phe2]VIP on the one hand, and of isoproterenol on the other hand, to decrease homologous responses was comparable to their potency for receptor occupancy and adenylate cyclase activation. PMID- 2547608 TI - Two genes encode the adenine nucleotide translocator of maize mitochondria. Isolation, characterisation and expression of the structural genes. AB - Southern blot hybridisation of maize (Zea mays) genomic DNA with a specific cDNA probe and sequence analysis of several cDNAs have revealed that the mitochondrial adenine nucleotide translocator (ADP/ATP carrier) is encoded by two genes, both of which are expressed in dark-grown coleoptile tissue. The structural genes (designated ANT-G1 and ANT-G2), encoding the two translocator polypeptides, have been isolated from a maize genomic library and characterised by DNA sequence analysis, mung bean nuclease mapping and primer extension. The two genes each contain two introns and encode very similar proteins. Transcripts from both genes have long (approximately 200 nucleotides) 5' untranslated leaders which are unusual in that they contain several ATG codons upstream of the initiation codon. Transcript analysis of RNA extracted from serial sections of developing maize leaves reveals that the translocator genes are most actively transcribed in the basal meristem, while transcripts were undetectable in green leaves. PMID- 2547609 TI - The genes for human alcohol dehydrogenases beta 1 and beta 2 differ by only one nucleotide. AB - Allelic differences at the alcohol dehydrogenase (ADH) and aldehyde dehydrogenase loci may have an important role in an individual's alcohol sensitivity. We have cloned and sequenced all nine exons of an ADH2(2) allele which codes for an 'atypical' ADH, ADH beta 2. Our sequence data shows that the histidine at residue 47 of ADH beta 2 is encoded by CAC. Surprisingly, no silent substitution was found between the coding regions of ADH2(1) [Duester, G., Smith, M., Bilanchone, V. & Hatfield, G. W. (1986) J. Biol. Chem. 261, 2027-2033.] and ADH2(2) alleles over the 1122 nucleotide sites. PMID- 2547610 TI - Inhibitory effect of nonviable preparations from human immunodeficiency virus 1 on inositol phospholipid metabolism. AB - Previously it was established [Pahwa, S., Pahwa, R., Saxinger, C., Gallo, R. C. & Good, R. A. (1985) Proc. Natl. Acad. Sci. USA 82, 8198] that nonviable preparations of human immunodeficiency virus 1 (HIV-1) abolish the proliferative response of human lymphocytes to phytohemagglutinin A. Now we describe that this effect might be, at least partially, due to an impairment of the function of phospholipase C. It was found that addition of HIV-1 preparation to lymphocytes diminished the stimulation of phosphatidylinositol phosphorylation caused by phytohemagglutinin A. Moreover, this preparation completely abolished the phytohemagglutinin-A-stimulated release of inositol trisphosphate and prevented a translocation of protein kinase C from cytosol to membranes. From this data we conclude that nonviable HIV-1 preparations inhibit the intracellular signalling pathway, leading to a reduced mitogenic response to phytohemagglutinin A, at the level of protein kinase C. PMID- 2547611 TI - Characterization of distinct mRNAs coding for putative isozymes of 6 phosphofructo-2-kinase/fructose-2,6-bisphosphatase. AB - Three distinct clones encoding full-length 6-phosphofructo-2-kinase (PFK 2)/fructose-2,6-bisphosphatase (FBPase-2) were characterized from a rat liver cDNA library. Clone 22c was 1859 bp long and coded for the 470 amino acids of the bifunctional subunit of the liver homodimer. This polypeptide is phosphorylated on serine 32 by cyclic-AMP-dependent protein kinase. Clone 4c (2681 bp) had a coding region identical to that of clone 22c but it included a putative intron of 959 bp. In clone 5c (1750 bp), the sequence upstream from amino acid 33 differed from that in clone 22c and coded for a unique N-terminal portion of 10 amino acids. Poly(A)-rich RNA from rat tissues was hybridized with cDNA probes corresponding to the unique N-terminal portions of clones 22c and 5c. Dot and Northern blots showed signals indicative of three distinct PFK-2/FBPase-2 mRNAs. There were a 6.8-kb mRNA typical of cardiac tissue, a 2.1-kb mRNA typical of liver, corresponding to clone 22c, and a 1.9-kb mRNA typical of skeletal muscle, corresponding to clone 5c. Primer extension analysis showed that clones 22c and 5c were nearly complete since their respective 5'-untranslated sequences were at most 96/97 bp and 44 bp shorter than the corresponding mRNAs. These data provide a molecular basis for the existence of PFK-2/FBPase-2 isozymes. PMID- 2547612 TI - Nucleotide sequence of cDNA coding for saporin-6, a type-1 ribosome-inactivating protein from Saponaria officinalis. AB - We have isolated and sequenced partial cDNA clones that encode SO-6, a ribosome inactivating protein from Saponaria officinalis. A cDNA library was constructed from the leaves of this plant and screened with synthetic oligonucleotide probes representing various portions of the protein. The deduced amino acid sequence shows the signal peptide and a coding region virtually accounting for the entire amino acid sequence of SO-6. The sequence reveals regions of similarity to other ribosome-inactivating proteins, especially in a region of the molecule where critical amino acid residues might participate in the active site. PMID- 2547613 TI - Secretory granules and endosomes show saltatory movement biased to the anterograde and retrograde directions, respectively, along microtubules in AtT20 cells. AB - AtT20 (clone D16V) cells develop long neurite-like processes in the growth cones of which secretory granules containing ACTH accumulate. These secretory granules have an acidic pH. Using acridine orange as a vital stain for acidic organelles, in combination with video-enhanced fluorescence microscopy, and subsequent immunolabeling with rabbit antibodies against ACTH, we have shown that these secretory granules move by saltations along the processes. During saltations velocities of 3 to 5 microns/s are achieved. The majority of the secretory granules move in the anterograde direction but some move retrogradely. The growth cones and processes are the site of extensive endocytosis. Using Lucifer Yellow as a vital stain we have shown that most endosomes move by saltations retrogradely. Movement of both secretory granules and endosomes is dependent upon microtubules. Individual secretory granules or endosomes never reverse the direction of their movement as they traverse the processes. Neutralization of the lumen of these acidic organelles with NH4Cl does not inhibit their movement or change its direction. PMID- 2547614 TI - Mannose 6-phosphate receptor in porcine thyroid follicle cells. Localization and possible implications for the intracellular transport of thyroglobulin. AB - Thyroglobulin has been shown to be phosphorylated and to carry the mannose 6 phosphate (M6P) signal in terminal position. In order to investigate whether the cation-independent mannose 6-phosphate receptor (CI-MPR) can possibly play a role in the transport of thyroglobulin the localization of the receptor was analyzed in thyroid follicle cells. The immunocytochemical observations showed that the CI MPR is primarily located in elements of the endocytic pathway such as coated pits and endosomes. This localization of the CI-MPR in thyrocytes differs from the receptor sites in other cell types by the rare occurrence of the CI-MPR in cisternae of the Golgi complex. The observations are interpreted as an indication that the relatively small amount of receptor in the Golgi complex might be occupied primarily by lysosomal hydrolases. The CI-MPR in thyrocytes might, therefore, be unable to bind and to convey thyroglobulin efficiently. The receptor is, however, a binding site for thyroglobulin at the apical plasma membrane and may, therefore, be involved in the binding of thyroglobulin and its transfer from the follicle lumen to lysosomes. PMID- 2547615 TI - A growth hormone-vesicular stomatitis virus G hybrid protein is rapidly degraded in lysosomes following transport to the cell surface. AB - We have expressed the hybrid protein, GHG3, in baby hamster kidney cells to study protein turnover. GHG3 contains the cytoplasmic and transmembrane domains of vesicular stomatitis virus G protein linked to the C-terminus rat growth hormone. Turnover of GHG3 was prevented by lysosomal inhibitors (leupeptin, chloroquine, primaquine or monensin), while the accumulated GHG3 was localized to intracellular vesicles, results indicating that degradation occurred in lysosomes. The kinetics of degradation at 34 degrees C were determined in pulse chase studies of metabolically labeled cells. After a lag period of 1 h, degradation was rapid (t1/2 = 1.25 h). The fate of GHG3 during the lag period was determined by immunofluorescence. We detected GHG3 on the cell surface when growth hormone antiserum was added to the growth medium 90 min prior to fixation and staining. No staining was observed if protein synthesis was inhibited with cycloheximide 90 min prior to the addition of growth hormone antiserum, a result indicating that GHG3 was rapidly removed from the cell surface. Unless the cells were pretreated with cycloheximide, antiserum was also detected in intracellular vesicles, which showed that GHG3 was endocytosed. These data indicate that a pool of GHG3 is transported rapidly to the cell surface, endocytosed and with little or no recycling directed to lysosomes for degradation. PMID- 2547616 TI - Ultrastructural localization of viral DNA in thin sections of herpes simplex virus type 1 infected cells by in situ hybridization. AB - We have used in situ hybridization at the ultrastructural level to localize non encapsidated and encapsidated herpes simplex virus type 1 (HSV-1) genomes in nuclei of infected rabbit fibroblasts. A biotinylated cloned subgenomic HSV DNA fragment was used as hybridization probe. The probe hybridized to the viral DNA accessible at the surface of Lowicryl sections was revealed by immunogold labeling. Non-encapsidated viral DNA was detected exclusively within the virus induced central region of 4 h to 17 h infected nuclei. Localization of the probe either near the nuclear envelope or within marginated host chromatin was found only on HSV DNA which was packaged into viral nucleoids. The use in parallel of in situ hybridization with specific staining for DNA and autoradiography after tritiated thymidine incorporation, followed by either conventional fixation of the cells or the nucleoprotein loosening procedure, indicated that non encapsidated viral DNA and marginated host chromatin formed two juxtaposed compartments without interpenetration even after experimentally produced mild dispersion of the nuclear components. PMID- 2547617 TI - Diet and reinfarction trial (DART): design, recruitment, and compliance. AB - A randomized controlled trial has been set up to examine the effect of diet on the secondary prevention of myocardial infarction, involving 2033 men. The trial has a factorial design, subjects being randomized independently to receive advice or no advice regarding three dietary factors: 1. a reduction in total fat and an increase in polyunsaturated fat intake; 2. an increase in fatty fish intake; 3. an increase in cereal fibre intake. Nearly half the men under 70 years of age who survived myocardial infarction during the recruitment period entered the trial, the commonest reason for exclusion being that the subject was already eating (or intended to eat) a diet which included one or more of the regimens being investigated. Detailed dietary questionnaires were completed by each subject after 6 months in the trial. The results suggest that compliance with the advice is reasonably good. The differences between the diets of the groups given and not given advice on fish and fibre were substantial; the difference attributable to advice on fat has been somewhat less than anticipated, partly because of failure to comply with advice and partly because of spontaneous changes in the diets of control subjects. PMID- 2547618 TI - 99mTc-labelling of leucocytes with 99mTc-DPO: a complex developed for myocardial imaging. AB - The lipophilic 99mTc-DPO complex, developed as a myocardial imaging radiopharmaceutical, was used to label leucocytes. After an incubation of 0.1 ml 99mTc-DPO (8 micrograms DMPE*2HCl) with mixed leucocytes in plasma, the labelling efficiency was over 70%. During incubation in 5 ml plasma, a loss of activity was found between 20% (1 h) and 35% (3 h) caused by elution. Disturbances of cell viability could not be found with the help of the chemiluminescence test. The in vivo recovery was determined in three dogs and was 45%-50% (0.5 h), 30%-36% (1 h), and 18%-24% (3 h). Autologous 99mTc-DPO-leucocytes were used on seven patients with suspected osteomyelitis, there were four true negative and three true positive results. The target/nontarget ratio determined by ROI in the positive cases was 1.8 to 2.5 at 3 h after injection. PMID- 2547619 TI - Adrenoleukodystrophy presenting as spinocerebellar degeneration. AB - The clinical features of 3 patients from a kindred with adrenoleukodystrophy and the analysis of their plasma sphingomyelin are described. Onset of symptoms was between the ages of 33 and 54 years. Ataxic gait and spasticity were the only symptoms noted during the early stage of the disorder. Dementia and optic atrophy were present in two of the cases. Baseline plasma cortisol was normal, but adrenocorticotropic hormone was elevated. Analysis of plasma sphingomyelin demonstrated an increase in very-long-chain (C24-C26) fatty acid. This study demonstrates that adrenoleukodystrophy may present with spinocerebellar symptoms. PMID- 2547620 TI - Evaluation of care for the disabled mentally ill: theoretical issues. AB - A precondition for the discharge from an institution of the chronically disabled mentally ill is the availability of adequate alternative facilities and services, optimally meeting their social and mental health needs and allowing as much quality of life as possible at the lowest possible cost. Only a few studies have found alternative care to lead to better outcomes than treatment in good-quality hospitals with respect to illness course and social disablement. Therefore, it is an important question whether extramural care and of what type is better and possibly less expensive than traditional inpatient treatment and for which patients. Health data collected at the national and regional levels provide a basis for continuous monitoring of utilization of services. Such data, however, usually have the limitation of being aggregate in nature and not including contacts with social services. These deficits can be avoided by using cumulative case registers, recording if possible all contacts of the socially disabled mentally ill with intramural and extramural mental health and social services and covering a define catchment area. Because the disabled mentally ill, depending on their own resources and those of their family and the community at large, have basic needs differing in each individual case, as well as various specific needs determined by the illness which they cannot meet on their own, it is necessary to provide a variety of facilities and services and to coordinate their work. Evaluation and cost-effectiveness investigations of complex packages of care yield generalizable results only if the forms of care under study are defined precisely enough.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547621 TI - Chemotherapy in non-small cell lung cancer (NSCLC). PMID- 2547622 TI - A comparison of cis-platinum-vindesine and cis-platinum-etoposide combined with radiotherapy for previously untreated localized inoperable non-small cell lung cancer. AB - Seventy-two previously untreated patients with localized inoperable non-small cell lung cancer were randomized to a study comparing the efficacy of cis platinum-vindesine (P-VDS) and of cis-platinum-VP16 (P-VP16), both combined with split-course radiotherapy. Fifty-nine patients were evaluable for response after the minimum requirement of two chemotherapy cycles. Both arms were further randomized to two split intervals, 3 or 5 weeks. The response rate to chemotherapy only (three cycles) was 66% for P-VDS and 50% for P-VP16. Radiotherapy increased the response rates to 83 and 67%, respectively. A Karnofsky score of 80% or more and the 3-week split interval were significant positive prognostic factors. Of all patients, 66% had local or combined recurrences and 17% relapsed at a distant site only. Since the 2-year survival rates are not strikingly better than those obtained by radiotherapy alone, we feel that these regimens should be restricted to further investigations of the role of chemotherapy in the treatment of different clinical presentations of NSCLC. PMID- 2547623 TI - Intracellular events associated with inhibition of B cell activation by monoclonal antibodies to HLA class II antigens. AB - We have investigated several aspects of the inhibitory effects of monoclonal antibodies (mAb) directed against MHC class II antigens in B cell activation/proliferation, using a panel of mAb specifically reactive with antigens encoded by HLA class II loci (DP, DQ, DR). All mAb except the anti-DP mAb inhibited significantly anti-mu plus B cell growth factor-induced DNA synthesis. Only one mAb, however, which was reactive with gene products of all three class II loci (DP, DQ, DR) inhibited anti-mu-induced DNA synthesis as well as c-myc mRNA expression. In addition, the same mAb inhibited the early events induced by anti-mu stimulation alone, including phosphatidylinositol turnover and elevation of [Ca2+]i. In contrast to previous findings in the murine system, none of the anti-MHC class II mAb used in this study increased the cAMP levels. PMID- 2547624 TI - Coordinated V gamma and V delta gene segment rearrangements in human T cell receptor gamma/delta+ lymphocytes. AB - Monoclonal antibodies (mAb) were used to characterize a panel (n = 46) of T cell receptor (TcR) gamma/delta+ T cell clones. Three of these antibodies have been described to react with specific variable region-encoded protein products and can therefore be used to detect functional gene rearrangements. The majority of peripheral blood-derived clones (43 out of 45) expressed the epitopes recognized by mAb BB3, encoded by the V delta 2 gene segment and mAb Ti gamma A, encoded by the V gamma 9 gene segment. These clones lacked the antigenic determinant recognized by mAb delta-TCS-1, encoded by the V delta 1 gene segment. The other two peripheral blood-derived clones and an ascites-derived clone were Ti gamma A , BB3- and delta-TCS-1+. Biochemical analysis revealed that all Ti gamma A+, BB3+ T cell clones expressed the disulfide-linked form of the receptor. The two peripheral blood-derived delta-TCS-1+ T cell clones expressed the nondisulfide linked form whereas the ascites-derived delta-TCS-1+ clone, AK119 expressed the disulfide-linked form of the TcR gamma/delta heterodimer. This indicates that V delta 1-encoded delta chains can be associated either with a C gamma 1- or a C gamma 2-encoded gamma chain. The preferential use of certain V gamma and V delta gene segments suggests the existence of a limited combinatorial diversity in TcR gamma/delta heterodimers, i.e. Ti gamma A+ (V gamma 9), BB3+ (V delta 2) and delta-TCS-1- disulfide-linked heterodimers and Ti gamma A-, BB3- and delta-TCS-1+ (V delta 1) disulfide- or non disulfide-linked forms. PMID- 2547626 TI - Mechanism of diuretic action of U-62,066E, a kappa opioid receptor agonist. AB - The mechanism of the diuretic action of U-62,066E, a highly selective kappa opioid agonist, was examined in unanesthetized rats and in isolated perfused inner medullary collecting ducts (IMCD). In Long-Evans rats, U-62,066E caused a dose-dependent increase in urine flow and a decrease in urine osmolality without affecting urinary excretion of Na+. The diuretic effect of U-62,066E was blocked by MR-2266, a kappa opioid receptor antagonist. U-62,066E showed no diuretic effect in homozygous hereditary diabetes insipidus rats (Brattleboro strain). In water-deprived rats, U-62,066E markedly inhibited plasma arginine vasopressin (AVP) levels through a kappa receptor-mediated mechanism. In rat IMCD perfused in vitro, 10(-5) M U-62,066E did not inhibit either the baseline or the AVP stimulated osmotic water permeability. We conclude that the inhibition of the release of AVP is the major if not the entire mechanism of the diuretic action of U-62,066E in rats. Although we ruled out the effect of this drug on the water permeability of IMCD, possible direct effects on other nephron structures remain to be established. PMID- 2547627 TI - Selective inhibition by nifedipine of the purinergic component of neurogenic vasoconstriction in the dog mesenteric artery. AB - The neurogenic contractions evoked by perivascular sympathetic nerve stimulation of dog mesenteric artery consist of purinergic and adrenergic components, and these components were selectively inhibited by alpha, beta-methylene ATP and prazosin, respectively. We examined the effects of Ca antagonists on both these components in dog mesenteric arteries. Nifedipine (10(-8)-10(-6) M) inhibited the purinergic and adrenergic contractions evoked by transmural electrical stimulation, and this inhibition was more evident for the purinergic component of the response. Nifedipine was also more potent to inhibit the contractile response to alpha, beta-methylene ATP than it was to inhibit the responses to noradrenaline. Verapamil and diltiazem also inhibited the purinergic and adrenergic responses induced by transmural electrical stimulation, alpha, beta methylene ATP or noradrenaline, but the extend of the inhibition was less than that seen with nifedipine. These three Ca antagonists had little effect on the 3H efflux evoked by electrical transmural stimulation of arteries that had been preincubated with [3H]noradrenaline. These results show that nifedipine is a selective inhibitor of the purinergic component of contractions evoked by sympathetic nerve stimulation of blood vessels. PMID- 2547625 TI - Chronic cocaine reduces alpha 2-adrenoceptor elicited mydriasis and inhibition of locus coeruleus neurons. AB - The effects of chronic cocaine (50 mg/kg per day for two weeks) administration on two alpha 2-adrenoceptor-mediated responses were studied in rats. Chronic administration of cocaine significantly (compared to sham controls) attenuated the alpha 2-adrenoceptor-mediated inhibition of noradrenergic locus coeruleus (LC) neurons as well as alpha 2-adrenoceptor elicited mydriasis. Noradrenergic LC neurons from the cocaine treated and sham sham groups differed significantly in their responsiveness to the inhibitory effects of clonidine (ED50 values micrograms/kg: sham 7.35 +/- 1.13 and cocaine-treated 17.17 +/- 4.40, P less than 0.05). The ED50 values for the mydriatic response were sham 5.71 +/- 0.49 and cocaine-treated 16.42 +/- 0.69 micrograms/kg, respectively, P less than 0.001. No differences in cardiovascular responses to systemically injected clonidine between the chronic cocaine- and sham-treated groups were observed. Chronic cocaine treatment attenuates the two alpha 2-adrenoceptor-mediated responses most likely via an interaction with central catecholaminergic neurotransmission. PMID- 2547628 TI - Gangliosides enhance behavioral and neurochemical effects induced by chronic desipramine (DMI) treatment. AB - Rats pretreated with gangliosides showed a significant enhancement of the anti immobility effect of desipramine (DMI) in the forced swimming test. Accordingly, an associated treatment of gangliosides and DMI for 7 days significantly enhanced beta-adrenergic down-regulation in the frontal cortex as compared with the effect of DMI alone. Gangliosides exerted an accelerating effect on the decrease of beta adrenoceptor density induced by DMI, since the down-regulation phenomenon appeared after 3 days of treatment. Gangliosides did not affect the pharmacokinetics of DMI, since associated acute or prolonged treatments did not modify the brain levels of DMI as compared to the levels of animals that had received DMI alone. These results evidence a stimulating effect of gangliosides on the development of adaptative receptor changes induced by chronic DMI treatment. PMID- 2547629 TI - Involvement of postjunctional purinergic mechanisms in the facilitatory action of bradykinin in neurotransmission in the rat vas deferens. AB - To investigate the nature of the bradykinin-induced potentiation of electrically driven muscle twitches in the isolated vas deferens, bradykinin, noradrenaline and adenosine 5'-triphosphate (ATP) concentration-response curves were made with control, reserpinized and chemically sympathectomized rats. Bradykinin potentiated the ATP- but not the noradrenaline-induced contractions in the epididymal and prostatic segments of the ductus. The epididymal segment of the ductus did not respond to transmural electrical stimulation following reserpine treatment. Bradykinin potentiated the muscular contractions caused by exogenous ATP but not by noradrenaline. In contrast, the transmurally evoked twitches of the prostatic portion of the ductus remained almost unaltered; bradykinin increased the motor effect of ATP without modifying the potency of noradrenaline. All neuronally induced contractile activity was absent in sympathectomized rats; bradykinin potentiated the contractile effect of ATP without altering the noradrenaline-induced contractions. These results suggest that bradykinin potentiates the ATP-evoked contractions by acting postjunctionally. PMID- 2547630 TI - Characterization of beta-adrenoceptor subtypes in rabbit mononuclear leukocytes. AB - Computer analysis of [125I]iodocyanopindolol competition studies using the relatively selective beta 1-adrenoceptor antagonist, ICI 89406, and the beta 2 selective antagonist, ICI 118551, on rabbit mononuclear leukocyte plasmalemmal preparations favored a two-site model indicating that both beta 1- and beta 2 adrenoceptor subtypes were present in approximately equal numbers. In contrast, similar studies performed on rabbit cardiac sarcolemma favored a one site model consistent with the presence of beta 1-adrenoceptors. Consequently, rabbit mononuclear leukocytes may provide a useful model for studying selective modulatory mechanisms of beta 1- and beta 2-adrenoceptors. PMID- 2547631 TI - Receptor-mediated stimulation of inositol phospholipid hydrolysis in human brain. AB - Stimulation of inositol phospholipid hydrolysis by transmitter receptor agonists was studied in slices from the occipital, temporal and cerebellar cortex that were surgically removed from three patients during ablation of brain tumors. Norepinephrine and the muscarinic acetylcholine agonist carbamylcholine increased the content of [3H]inositol monophosphate (InsP) in all the regions examined, whereas the glutamate receptor agonist quisqualate was effective only in the cerebellar slices. These results provide evidence that neurotransmitter receptors are coupled to inositol phospholipid hydrolysis in human brain tissue. PMID- 2547632 TI - Glycine reverses 7-Cl kynurenate blockade of glutamate neurotoxicity in cell culture. PMID- 2547633 TI - Effects of adenosine analogues on force and cAMP in the heart. Influence of adenosine deaminase. AB - The effects of the adenosine receptor agonists (-)-N6-phenylisopropyladenosine (PIA) and 5'-N-ethylcarboxamideadenosine (NECA) on the force of contraction, adenylate cyclase activity and cAMP content in the presence of isoprenaline (Iso) were studied in ventricular preparations of the guinea-pig heart. Only in the presence of adenosine deaminase (ADA) and 50 mM sodium chloride, i.e. under 'optimal' conditions, did PIA and NECA reduce the Iso-stimulated adenylate cyclase activity in broken cell preparations, with a maximal effect of about 25%. In electrically driven (1 Hz) papillary muscles from guinea-pigs, both compounds concentration dependently reduced the Iso-stimulated force of contraction maximally by about 50% in the presence of ADA (1 microgram/ml). cAMP was measured in the same preparations. Low concentrations (0.1-1 microM) of PIA reduced the cyclic AMP content while higher concentrations increased the cyclic AMP content. The negative inotropic effect of NECA was accompanied by a concentration dependent increase in the cyclic AMP content. We conclude that the negative inotropic effect of PIA in the presence of Iso is only in part due to a decrease in the cyclic AMP content resulting from inhibition of adenylate cyclase activity. Such an effect was only detected in the presence of ADA so that endogenous adenosine can obviously mask small effects of PIA on adenylate cyclase activity or the cyclic AMP content. In addition, the negative inotropic effect of NECA in the presence of isoprenaline was not accompanied by a reduction but an increase in the cyclic AMP content.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547634 TI - Alpha-adrenergic regulation of action potentials in isolated rat cardiomyocytes. AB - Radioligand binding studies and studies in which inositol phosphate levels were assayed have provided substantial evidence for the existence of alpha 1 adrenoceptors in isolated rat ventricular myocytes, a pure cardiac preparation free of neuronal and vascular elements. Since correlational electrophysiological studies have not been carried out in this model, we investigated alpha-adrenergic effects on action potentials using intracellular microelectrodes to impale the myocytes. Epinephrine, in the presence of propranolol (10(-6) M), rapidly increased the action potential duration in a concentration-dependent manner (threshold concentration of 30 nM, EC50 of 100 nM). The maximal upstroke velocity and overshoot of Ca2+-mediated action potentials (15 mM K+) were also increased. Epinephrine did not significantly affect the resting membrane potential. The alpha 1-adrenoceptor antagonist, prazosin (10(-7) M) blocked epinephrine's effects. LiCl (10 mM), an inhibitor of inositol-phosphate phosphatases potentiated the effects of epinephrine (30 nM). The results suggest that the isolated rat cardiomyocyte is a suitable preparation for examining the ionic and molecular mechanisms of direct alpha-adrenergic effects on the cardiac membrane. PMID- 2547635 TI - Local anesthetics: lipophilicity, charge, diffusion and site of action in isolated neuron. AB - Sodium channel blocking activity was measured in a series of newly synthesized tertiary amine compounds and their quaternary derivatives applied externally on internally in the rat sensory neuron. The large difference in effectiveness of the quaternary compounds on external and or internal application became smaller and even disappeared with increasing lipophilicity of the compounds. No such difference was observed in the tertiary analogs. It was concluded that lipophilicity played an important role in determining the channel blocking activity of externally applied quaternary compounds. The results suggest that, in addition to the neutral form, highly lipophilic amine local anesthetics may penetrate into and/or pass across the neuronal membrane probably as electroneutral ion pair complexes consisting of the cationic form and an appropriate anion. The present results support our hypothesis which stresses, in addition to dissociability, the role of lipophilicity of amine local anesthetics in the pH dependence of their effect. PMID- 2547636 TI - Characterization of the effects of the acute and repeated administration of MK 801 on the release of adrenocorticotropin, corticosterone and prolactin in the rat. AB - In addition to its producing profound changes in behavior, phencyclidine (PCP) disrupts neuroendocrine function in the rat. Because PCP binds to PCP as well as sigma receptors, it is not known which receptor type mediates the various effects of the drug. The purpose of the present study was to characterize the effects of the acute administration of enantiomers of MK-801, a compound with a high degree of selectivity for PCP over sigma receptors, on the release of ACTH, corticosterone and prolactin in the rat. In addition, MK-801 was administered daily for eight days in order to test whether tolerance develops to MK-801 induced ACTH and corticosterone release after repeated administration. While both enantiomers of MK-801 markedly increased plasma levels of ACTH and corticosterone, the (+) enantiomer was more potent. Tolerance developed to MK-801 induced increases in ACTH and corticosterone after repeated administration. Plasma prolactin levels were not affected by either the acute or the repeated administration of MK-801. These results suggest that the decrease in plasma levels of prolactin produced by PCP-like drugs is not mediated by PCP receptors, and may be a marker for a sigma receptor-mediated effect. PMID- 2547637 TI - Diazepam enhances the inhibitory action of adenosine on transmission at the frog neuromuscular junction. AB - The effect of diazepam and its interaction with adenosine on evoked endplate potentials (e.p.p.s) and on twitch tension were investigated in innervated sartorius muscles of the frog. Diazepam (100 microM) reversibly decreased the amplitude of the e.p.p.s and the twitch responses evoked by indirect stimulation, and reversibly increased the resting membrane potential recorded from the endplates. In a concentration (30 microM) virtually devoid of an effect on the e.p.p.s, twitch responses, or resting membrane potential of the muscle fibres, diazepam potentiated the inhibitory action of adenosine on neuromuscular transmission, but not that induced by the stable analogue of adenosine 5'-N ethylcarboxamide adenosine (NECA), which is not a substrate for the adenosine uptake system. The potentiating effect of diazepam was not observed in the presence of dipyridamole, an adenosine uptake blocker which potentiated the effect of adenosine on neuromuscular transmission. Diazepam shifted to the left the concentration-response curve obtained for adenosine in the presence of the adenosine receptor antagonist 8-phenyltheophylline (8-PT). The results suggest that diazepam acts at the frog neuromuscular junction by increasing the level of adenosine at the junction level; this increase probably results from inhibition in the uptake of the nucleoside. PMID- 2547638 TI - Benzofuran analogues of baclofen: a new class of central and peripheral GABAB receptor antagonists. AB - Two novel beta-(benzo[b]furan) analogues of baclofen (4-amino-3-benzo[b]furan-2 ylbutanoic acid, 9G, and 4-amino-3-(5-methoxybenzo[b]furan-2-yl)butanoic acid, 9H) antagonised the baclofen-induced depression of twitch contractions in the guinea-pig isolated ileum (estimated apparent pA2 3.9 and 4.1 respectively); both 9G and 9H also antagonised in a dose-dependent manner the baclofen-induced reduction of repetitive paroxysmal discharges in rat neo-cortical slice preparations maintained in Mg2+-free Krebs solution. These benzofurans evidently represents a new class of GABAB-receptor antagonist. PMID- 2547639 TI - Increases in vascular alpha 1-adrenoceptor affinity and PI turnover in DOCA-salt hypertensive rats. AB - The pD2 value for norepinephrine and the pA2 value for prazosin in mesenteric larger resistance vessels from deoxycorticosterone acetate (DOCA)-salt hypertensive rats were significantly higher than the values of normotensive rats, suggesting that alpha 1-adrenoceptors have a higher affinity in hypertensive rats. The accumulation of [3H]inositol monophosphate induced by norepinephrine was significantly greater in the mesenteric artery from hypertensive rats than in the mesenteric artery from normotensive rats. These results suggest that the enhanced phosphatidylinositol turnover seen in DOCA-salt hypertensive rats may contribute, at least partly, to the higher affinity of alpha 1-adrenoceptors. PMID- 2547640 TI - Muscarinic receptors in human airway smooth muscle are coupled to phosphoinositide metabolism. AB - In the present study we investigated whether muscarinic receptors in human airway smooth muscle are coupled to phosphoinositide metabolism as a possible transduction mechanism of contraction. Using isolated bronchial smooth muscle preparations, we found that the muscarinic agonist methacholine caused a time- and concentration-dependent accumulation of inositol phosphates in the presence of lithium, an effect which could be inhibited by atropine. Apart from its physiological significance, this finding may have great relevance for the biochemical investigation of cholinergic hyperresponsiveness in the airways of asthmatic patients. PMID- 2547641 TI - Isoniazid, an inhibitor of GABAergic transmission, enhances [35S]TBPS binding in rat cerebral cortex. AB - Isoniazid, administered to rats one hour before killing produced a dose-dependent enhancement of [35S]t-butylbiciclophosphorothionate ([35S]TBPS, 2 nM) measured ex vivo in unwashed membrane preparation of the cerebral cortex. Saturation experiments revealed that the effect of isoniazid was due to an increase (+36%) in the total number of [35S]TBPS binding sites. Diazepam (3 mg/kg i.p.) administered 15 min after isoniazid antagonized the enhancement of [35S]TBPS binding elicited by isoniazid. Moreover, diazepam itself induced a significant decrease (-30%) in the total number of [35S]TBPS binding sites. These results provide the first direct evidence that 'in vivo' alterations in the function of the GABA-dependent chloride channel can be detected in vitro by studying the binding of [35S]TBPS to its recognition sites in the GABAA receptor complex. Our finding suggests a new model suitable to study biochemically the function of GABAergic synapses under various physiological and pharmacological conditions. PMID- 2547642 TI - Intracellular pH regulation in basal corneal epithelial cells measured in corneal explants: characterization of Na/H exchange. AB - Intracellular pH (pHi) was measured in basal corneal epithelial cells from fresh corneal explants using the pH sensitive fluorescent dye 2',7'-bis(2-carboxyethyl) 5(6)-carboxyfluorescein (BCECF). The overlying superficial and wing cells were removed by mechanical scraping to expose basal cells attached to their basal lamina. Tissue pieces with attached, dye-loaded basal cells were mounted in a microscope-stage-perfusion chamber which allowed rapid changes of Ringer's bathing solutions while measuring BCECF fluorescence. In NaCl-Ringer's (bicarbonate free). pHo 7.40, resting cell pHi was 7.34 +/- 0.03 (+/- S.E.M., n = 31). Buffering capacity measured by NH4Cl treatment was 31 mM pH at pHi 7.34 and increased with decreasing pHi. Recovery from 20 mM NH4Cl-induced acid loads was dependent on the presence of Na and inhibited by 1 mM amiloride. Adding amiloride to resting cells caused a slow, reversible acidification (0.04 pH units min-1). These results indicate the presence of Na:H exchange, its role in responding to acid loads and in maintaining resting cell pHi. Activation of Na:H by Nao showed simple saturation kinetics, with Km = 44 mM. Net proton efflux via Na:H exchange increased with decreasing pHi and was enhanced by depleting cells of Nai, suggesting roles for both pHi and Nai in control of Na:H activation. PMID- 2547643 TI - Angiotensin converting enzyme (ACE) in iris vessels. An ultrastructural study. PMID- 2547644 TI - Non-identity of cGMP as the guanine nucleotide stimulated to bind to ROS by light and ATP. AB - Light, in the presence of ATP, has been reported to stimulate cGMP binding to a 58 kDa protein in ROS (rod outer segments, Fesenko and Krapivinsky, 1986b, Photobiochem. Photobiophys. 13 345-58). This apparent light-related redistribution of ROS cGMP has been suggested to eliminate any requirement for phosphodiesterase-promoted hydrolysis of cGMP in the mechanism subserving phototransduction. Using conditions identical to those previously reported, this effect of light and ATP was examined further by characterizing the metabolic products that arise and the nucleotides that become liganded. The increased binding of radiolabeled guanine nucleotide upon illumination of ROS in the presence of ATP was confirmed, but the species of guanine nucleotide that were stimulated to bind under these conditions were identified as [32P]GDP and [32P]GTP rather than [32P]cGMP. The precautions to prevent enzymic hydrolysis of cGMP, which included conducting the reactions at 0 degrees and the addition of 3 isobutyl-l-methylxanthine (250 microM) to the reaction mixture did not prevent about a 20-fold increase in the rate of phosphodiesterase-catalyzed hydrolysis of radiolabeled cGMP by light when ATP was also present. This stimulation of phosphodiesterase activity is undoubtedly related to transphosphorylation by exogenous ATP of endogenous GMP and GDP involving catalytic actions of guanylate kinase and nucleoside diphosphate kinase in isolated ROS. These enzymes can also serve to generate [32P]GDP and [32P]GTP, which subsequently bind to ROS components. Such a mechanism involving ATP as phosphoryl donor was supported by observing that an analog of ATP (beta,gamma-methyleneadenosine 5'-triphosphate), which cannot serve as a phosphoryl donor, did not increase radiolabeled guanine nucleotide binding. Although several ROS proteins can form filter-retainable complexes with GDP and GTP, the properties of the 58 kDa protein found to be photoaffinity labeled with radioactive guanine nucleotide are most characteristic of those attributable to tubulin. The previous report that illumination in the presence of ATP stimulates the binding of cGMP to ROS components finds no support from the data obtained in the present studies. PMID- 2547645 TI - Superoxide anion scavenging effect and superoxide dismutase activity of Ginkgo biloba extract. AB - Ginkgo biloba extract is known to be efficient in diseases associated with free radical generation. The purpose of this work was to study, under in vitro conditions, the action of Ginkgo biloba extract (Gbe) against superoxide anion (O2-.), which is directly or indirectly implicated in cell damage. Gbe appears to have both an O2-. scavenging effect and also a superoxide dismutase activity. Its antiradical effect was demonstrated by low temperature electron spin resonance and in a non-enzymatic system (phenazine methosulfate-NADH), and its enzymatic activity was shown by polarographic determination. PMID- 2547646 TI - GABA-A-mediated gastrin release induced by baclofen in the isolated vascularly perfused rat stomach. AB - In order to investigate the role of peripheral GABA-B receptors, the effects of the putative GABA-B agonist baclofen on immunoreactive gastrin release from an isolated vascularly perfused rat stomach preparation were examined. The vascular infusion of baclofen at graded concentrations induced a dose-dependent increase in gastrin release; this was unaffected by the GABA-B antagonist delta aminovaleric acid, but was fully prevented by the selective GABA-A antagonist bicuculline as well as by atropine or tetrodotoxin. These results suggest that the stimulant effects of baclofen are mediated by nervous cholinergic structures associated with GABA-A receptors, and indicate that this GABA-B agonist must be regarded as a partial agonist of peripheral GABA-A receptors. PMID- 2547647 TI - The molecular genetics of human lung cancer. AB - With the development of molecular biological techniques the search for genetic alterations in cancer cells has resulted in the beginning of a molecular description of cellular transformation. Most of these genetic changes occur in genes which have a role in the control of cellular growth and development, the proto oncogenes. In the last decade, it has become clear that the myc and ras oncogene families are important in the carcinogenesis of human lung cancers. The myc oncogenes are usually found to be altered in small cell lung cancer (SCLC), and these alterations appear to correlate with rapid growth and progression. Mutations in the Kras gene are specific for adenocarcinoma, a subclass of non small cell lung cancer (NSCLC). Kras gene mutations are closely associated with tobacco smoking, since all were found in adenocarcinomas from patients with a history of smoking. The erbB oncogene, which encodes the epidermal growth factor receptor, is often highly expressed in epidermoid carcinomas. The roles for other oncogenes, such as raf or myb, as well as those of "suppressor" genes remain to be investigated, but may be of paramount importance. The study of alterations in proto oncogenes may aid in the (sub)classification and diagnosis of lung cancer, and may yield useful prognostic information in the near future. PMID- 2547648 TI - The effect of heparin on the inositol 1,4,5-trisphosphate receptor in rat liver microsomes. Dependence on sulphate content and chain length. AB - Heparin is known to inhibit the binding of inositol 1,4,5-trisphosphate (Ins 1,4,5-P3) to high-affinity binding sites and to inhibit Ins 1,4,5-P3-induced Ca2+ release from intracellular membrane-bound stores [(1987) J. Biol. Chem. 262, 12132-12136; (1987) FEBS Lett. 228, 57-59]. We have performed studies to clarify the structural requirements for this action of heparin in rat liver microsomes. Both N- and O-linked sulphate groups contribute to binding activity, since de-N sulphated heparin was without effect on the Ins 1,4,5-P3 receptor whereas a polyxylan bearing only O-linked sulphates (pentosan polysulphate) was as active as heparin. Therefore, the density of negative charge contributed by sulphate groups is important for the binding of heparin. Heparins with high and low affinity for antithrombin III both inhibited Ins 1,4,5-P3 binding. There was a strong dependence on chain length, since binding activity decreased dramatically as the size of the heparin chain was reduced below that of 18-24 monosaccharide units. PMID- 2547649 TI - Electron paramagnetic resonance evidence that cellular oxygen toxicity is caused by the generation of superoxide and hydroxyl free radicals. AB - Cells require molecular oxygen for the generation of energy through mitochondrial oxidative phosphorylation; however, high concentrations of oxygen are toxic and can cause cell death. A number of different mechanisms have been proposed to cause cellular oxygen toxicity. One hypothesis is that reactive oxygen free radicals may be generated; however free radical generation in hyperoxic cells has never been directly measured and the mechanism of this radical generation is unknown. In order to determine if cellular oxygen toxicity is free radical mediated, we applied electron paramagnetic resonance, EPR, spectroscopy using the spin trap 5,5'-dimethyl-1-pyrroline-N-oxide, DMPO, to measure free radical generation in hyperoxic pulmonary endothelial cells. Cells in air did not give rise to any detectable signal. However, cells exposed to 100% O2 for 30 min exhibited a prominent signal of trapped hydroxyl radical, DMPO-OH, while cell free buffer did not give rise to any detectable radical generation. This cellular radical generation was demonstrated to be derived from the superoxide radical since the observed signal was totally quenched by superoxide dismutase, but not by equal concentrations of the denatured enzyme. It was confirmed that the hydroxyl radical was generated since in the presence of ethanol the CH3 CH(OH) radical was formed. Loss of cell viability as measured by uptake of trypan blue dye was observed paralleling the measured free radical generation. Thus, superoxide and hydroxyl radicals are generated in hyperoxic pulmonary endothelial cells and this appears to be an important mechanism of cellular oxygen toxicity. PMID- 2547650 TI - Na+/H+ exchange is not necessary for protein kinase C-mediated effects in platelets. AB - The role of Na+/H+ exchange in protein kinase C-mediated effects in platelets was investigated by studying the effect of removal of extracellular Na+ ([Na+]e) on the different responses induced by phorbol 12-myristate 13-acetate (PMA) and 1,2 dioctanoylglycerol (diC8). None of the responses studied, namely, protein phosphorylation, translocation of enzyme activity to the membrane fraction, potentiatory and inhibitory effect on platelet activation ([Ca2+]i, arachidonate and granule release) showed an absolute dependence on [Na+]e. With the exception of dense-granule release, which was clearly potentiated by the removal of [Na+]e and showed a negative correlation with exchanger activity, the other effects of PMA and diC8 were not affected by [Na+]e removal. It is concluded that Na+/H+ exchange is not essential for protein kinase C activation in platelets. PMID- 2547651 TI - Two early genes of bacteriophage T5 encode proteins containing an NTP-binding sequence motif and probably involved in DNA replication, recombination and repair. AB - It is demonstrated, by computer-assisted analysis, that T5 bacteriophage early genes D10 and D13 encode proteins containing the purine NTP-binding sequence motif. The D10 gene product is shown to be a member of a recently characterized superfamily of (putative) DNA and RNA helicases. The D13 gene product is related at a statistically significant level, to the gene 46 product of bacteriophage T4 which is a component of an exonuclease involved in phage DNA replication, recombination and repair. A lower but also significant degree of sequence similarity was detected between the gene D12 product of T5 and the gene 47 product of T4, the second component of the same nuclease. It is hypothesized that both D10 and D13 gene products of T5 might be NTPases, possibly DNA-dependent, mediating NTP-consuming steps during phage DNA replication, recombination and/or repair. PMID- 2547652 TI - Energy profiles in the acetylcholine receptor (AChR) channel. The MII-helix model and the role of the remaining helices. AB - It is demonstrated by theoretical computations that no favorable energy profile for cation transfer can be obtained in a model of the AChR channel constructed with the sole five MII helices of the inner wall. A favorable profile is obtained upon including the effect of the remaining helices of the five subunits. The decisive role, for the exit of the ion, of the charged residues situated at the N terminal of the MII segments, established before, is underlined further. The role of the other elements of the channel wall (peptide carbonyl oxygens, hydrocarbon residues and polar side chains) is analyzed. PMID- 2547653 TI - The testicular transcript of the angiotensin I-converting enzyme encodes for the ancestral, non-duplicated form of the enzyme. AB - The endothelial angiotensin I-converting enzyme (ACE) is organized in two large homologous domains, each bearing a putative active site. However, only one of these sites is probably involved in catalyzing the conversion of angiotensin I into angiotensin II. The testicular form of ACE is equally active, encoded by the same gene, but translated from a shorter mRNA. Molecular cloning of the human testicular ACE cDNA indicates that the mRNA codes for 732 residues (vs 1306 in endothelium). The testicular transcript corresponds to the 3' half of the endothelial transcript and encodes one of the two homologous domains of endothelial ACE, preceded by a short specific sequence. This 5' specific sequence contains 228 nucleotides and encodes 67 amino acids, including the putative signal peptide followed by a serine/threonine-enriched region, presumably glycosylated. The testicular transcript corresponds to the ancestral, non duplicated form of the ACE gene. Since the carboxyl-terminal domain of the endothelial ACE is expressed in the testicular enzyme, it is likely that it bears the active site in both forms. PMID- 2547654 TI - P2-purinergic agonists activate phospholipase C in a guanine nucleotide- and Ca2+ dependent manner in FRTL-5 thyroid cell membranes. AB - Various adenine nucleotides activated phospholipase C of FRTL-5 cell membranes in the following order of activity, ATP gamma S greater than ATP greater than AppNp greater than AppCp = ADP greater than MeSATP. This order was well consistent with that observed in intact cells. Such activation occurred only in the presence of appropriate concentrations of GTP gamma S and Ca2+, in a way similar to the norepinephrine-induced activation. NaF, a non-specific GTP-binding protein (G protein) activator, also stimulated the enzyme. These adenine nucleotides, norepinephrine and NaF-induced activations were inhibited by GDP beta S. We conclude that a G-protein is involved in the adenine nucleotides-induced activation of phospholipase C via P2-purinergic receptor in FRTL-5 cells. PMID- 2547655 TI - Three apparent receptor subtypes for the endothelin/sarafotoxin family. PMID- 2547656 TI - Purification of the mitochondrial carnitine carrier by chromatography on hydroxyapatite and celite. AB - The carnitine carrier from rat liver mitochondria has been extracted with Triton X-100 ad partially purified by chromatography on hydroxyapatite and celite. During purification the activity of the carrier was monitored by functional reconstitution into liposomes. The purified fraction is 250-fold enriched with respect to the N-ethylmaleimide-sensitive carnitine/carnitine transport activity. The substrate specificity and the inhibitor sensitivity of carnitine transport in liposomes resemble closely those described for the transport of carnitine in mitochondria. PMID- 2547658 TI - Pulse radiolysis study of the reactivity of Trolox C phenoxyl radical with superoxide anion. AB - The reaction between the phenoxyl radical of Trolox C, a water-soluble vitamin E analogue, and superoxide anion radical was examined by using the pulse radiolysis technique. The results indicate that the Trolox C phenoxyl radical may undergo a rapid one-electron transfer from superoxide radical [k = (4.5 +/- 0.5) x 10(8) M 1.S-1] to its reduced form. This finding indicates that superoxide radical might play a role in the repair of vitamin e phenoxyl radical. PMID- 2547657 TI - Mu type opioid receptors in rat periaqueductal gray-enriched P2 membrane are coupled to G-protein-mediated inhibition of adenylyl cyclase. AB - The periaqueductal gray (PAG) region of the midbrain has been implicated in both stimulation-produced and opioid-induced analgesia. High-affinity mu-selective opioid-binding sites associated with mu type opioid receptors have been detected in rat PAG-enriched P2 membranes, and these receptors have been shown to be coupled to guanine nucleotide-binding proteins (G-proteins). In the present study the potential G-protein-mediated coupling of mu type opioid receptors to the inhibition of adenylyl cyclase was examined utilizing in vitro adenylyl cyclase assays. In the presence of Na+, opioid agonists inhibited adenylyl cyclase in a mu selective, naloxone reversible, dose dependent, and pertussis toxin sensitive manner. Overall the data suggests that mu type opioid receptors in the rat PAG are coupled to G-protein-mediated inhibition of adenylyl cyclase. PMID- 2547660 TI - Regulation of DNA supercoiling in Escherichia coli: genetic basis of a compensatory mutation in DNA gyrase. AB - Bacterial DNA supercoiling is controlled by balancing the supercoiling activity of DNA gyrase and the relaxing activity of DNA topoisomerase I. We have characterized the gyrB gene from a top A deletion mutant of Escherichia coli (DM800) that has a compensatory mutation in gyrB, lowering the activity of gyrase 10-fold, and thereby redressing the intracellular level of supercoiling. The mutant gene differs from the wild type in carrying three rather than two direct tandem repeats of a 6 bp sequence encoding Ala-Arg. We suggest this novel mutation affects domain spacing and was generated by an unequal crossing over event, possibly involving gyrase. PMID- 2547659 TI - Light chain of tetanus toxin intracellularly inhibits acetylcholine release at neuro-neuronal synapses, and its internalization is mediated by heavy chain. AB - The ability of the two-chain form of tetanus toxin (TeTx), its constituent light (LC) or heavy (HC) chains, and papain fragment to block evoked acetylcholine (ACh) release in the buccal ganglia of Aplysia californica was studied electrophysiologically. Extracellularly applied, TeTx or its B fragment (consisting of LC and beta 2, the amino-terminal portion of HC) blocked ACh release, whereas LC, HC, or the beta 2 fragment did not affect it. Toxicity was restored when LC was bath applied together with HC or the beta 2 fragment. When injected into the presynaptic neuron, TeTx, the B fragment or LC, but not HC, induced inhibition of ACh release. These results indicate that the blockade of ACh release by TeTx is mimicked by intracellular action of LC, the internalization of which is mediated by the HC via its amino-terminal moiety. PMID- 2547661 TI - Clinicopathological analysis of synchronous multiple gastric carcinoma. AB - Clinicopathological analysis was performed on 839 cases surgically resected for gastric carcinoma. The incidence of multiple gastric carcinoma was 4.8% (40 cases, 97 lesions). Multiple carcinoma was more frequently observed in early than in advanced carcinoma (P less than 0.01). The rate of intestinal type lesions was significantly (P less than 0.01) higher in multiple than in single gastric carcinoma, and all of the intestinal type carcinoma correlated with intestinal metaplasia, which is assumed to be closely related to pyloric and atrophic fundic gland area. Eight cases (20.0%) of multiple carcinoma were both in the upper one third and lower one-third of the stomach. Twenty-nine (51.9%) of the accessory lesions were not detected pre-operatively; 12 (21.1%) of them were detected only by postoperative histology. Twelve (48.0%) of 25 early cancerous foci located in the anterior wall and greater curvature were overlooked before operation. These results indicate that the whole stomach must be carefully examined to detect accessory carcinoma before gastric surgery, especially for intestinal type carcinoma, with greater attention paid to the anterior wall and greater curvature, and that complete removal of the pyloric and atrophic fundic gland area would be required for distal gastrectomy. PMID- 2547662 TI - In situ carcinoma of the breast. AB - The optimal management of ductal carcinoma in situ of the breast is controversial. With the introduction of the National Mammographic Breast Screening Programme the condition will be encountered more frequently. We have reviewed 76 patients from a 12-year period treated by one surgeon (R.W.B.) at the Nottingham City Hospital. Fifty-nine patients had either ductal carcinoma in situ or lobular carcinoma in situ; 17 patients had Paget's disease. The mean age at diagnosis was 54 years and the commonest mode of presentation was with a palpable breast lump. Pre-operative mammography was performed in 31 patients with ductal carcinoma in situ and 28 were reported as showing malignancy. Patients with a lesion in the breast parenchyma were treated either by mastectomy (simple, subcutaneous or 'wedge'--see text) or by lumpectomy and radiotherapy. Patients with Paget's disease were treated by simple mastectomy, wedge mastectomy or a cone excision of the nipple and underlying tissue. The mean follow-up period was 65 months. Patients treated by any of the procedures less than simple mastectomy had a significant chance of developing local recurrence. A detailed histological review was made and grade, microinvasion, calcification, necrosis and completeness of excision were assessed for each tumour. None of these factors was correlated with subsequent local recurrence. PMID- 2547663 TI - Influence of T and N stages on long-term survival in resectable small cell lung cancer. AB - The influence of T and N stages on long-term survival was evaluated in 49 patients with small cell lung cancer treated between 1975 and 1982. Adjuvant chemotherapy was administered in 34 patients and consisted of cyclophosphamide, doxorubicin and vincristine or etoposide, while 15 patients were treated with surgery alone. To date, overall median survival is 12 months, with nine patients (18.4%), who received adjuvant chemotherapy, currently alive and disease-free at 60 to 150 months. Survival was statistically influenced by the N stage but not the T stage; patients at the N0 stage had a significant increase when compared to N1 and N2 patients, on both 5-year survival (p less than 0.05 and p less than 0.025, respectively) and long-term survivors (p = 0.006 and p = 0.004, respectively). Results suggest that surgery and adjuvant chemotherapy might be an effective form of treatment in small cell lung cancer for T1, T2 and also T3 tumours without hilar or mediastinal lymph node involvement. PMID- 2547664 TI - Chemotherapy plus adjuvant surgery for local small cell lung cancer. AB - Eight patients with local Stage II (T2N1) and III (T3N0, T3N1, T2N2) small cell lung cancer received combination chemotherapy prior to elective surgery to assess the effectiveness of such a regimen in improving operability, preventing local relapse and extending survival. The regimen was well tolerated and prevented local relapse. However, the median time to recurrence of disease was 10 months and the median survival time 13 months, results which are similar to those achieved with chemotherapy and radiotherapy. Distant metastases, particularly in the brain, occurred predictably indicating that successful adjuvant surgery, despite preventing local relapse, may not afford additional survival benefit with currently available drug regimens. PMID- 2547665 TI - Serological markers in the diagnosis of histopathological types of nasopharyngeal carcinoma. AB - The titres of IgA against Epstein-Barr virus, viral capsid antigens and the titres of IgG against early antigen were found to be useful markers for the diagnosis of different histopathological types of nasopharyngeal carcinoma. PMID- 2547666 TI - Metastatic vipoma arising from colonic primary tumour. AB - We describe a case of a tumour of the sigmoid colon with hepatic metastases in a patient with previously documented ulcerative colitis. A diagnosis of metastatic vipoma was made on the basis of high plasma levels of vasoactive intestinal polypeptide (VIP). Profuse diarrhoea and profound metabolic upset were corrected by the use of a somatostatin analogue SMS 201-995, whilst conventional cytotoxic therapy produced a significant tumour response with return of the plasma VIP level to normal. PMID- 2547667 TI - [Analysis of the nature of responses of neurons of rat spinal ganglia evoked by the activation of GABA-B receptors]. AB - The GABA application evoked fast picrotoxin-sensitive depolarization as well as slow depolarization with decreasing membrane conductivity and hyperpolarization with increasing membrane conductivity in the neurons of the rat dorsal root ganglion. The slow picrotoxin-resistant responses to GABA were initiated with phenibut. The phenibut effects were modulated by cAMP concentration and inhibited by the pertussis toxin. The reversal potential of the phenibut effects was equal to the potassium equilibrium one. The phenibut effects were reduced by increasing K+ and Ca+ extracellular concentration, and enhanced in presence of trifluoroperazine. PMID- 2547668 TI - [Effect of niflumic acid on the components of inactivation of the sodium channels of the Ranvier node membrane]. AB - In voltage clamp experiments on the frog Ranvier node, niflumic acid did not alter fast or slow inactivation time courses in the wide range of membrane potentials but reduced the amplitude of the fast phase of inactivation. Fast and slow currents, corresponding respectively to fast and slow phases of inactivation, reversed at the same voltage, revealed different activation- and inactivation-voltage dependences both in intact fibre and after application of niflumic acid. The latter induced a shift of the steady-state inactivation curves for both components of inactivation towards more negative potentials without changing their steepness. The validity of suggestion of the existence of two populations of sodium channels in nerve membrane, is discussed. PMID- 2547669 TI - [Effect of bilateral renal ischemia on the sodium, potassium ATPase activity of the kidneys and erythrocytes in rats]. AB - The renovascular hypertension in the Wistar rats was induced by clipping both renal arteries. Within 3-4 weeks arterial pressure (AP) increased to maximal values. According to AP, the rats were divided into two groups: with AP less than or equal to 170 mm Hg and greater than 170 mm Hg. In 5-6 or 10-11 weeks the animals were decapitated and the Na,K-ATPase activities in the wholesome erythrocytes, their ghosts, and the cortex and medulla of kidneys were studied. Changes of the enzyme activity were only found in the medulla. A decreased enzyme activity (-20%) was revealed in the rats with AP less than or equal to 170 mm Hg within 5-6 weeks after renal ischemia, the activity increasing again in 10-11 weeks after the operation. The decreasing of Na,K-ATPase activity within 5-6 weeks after renal ischemia could be a mechanism of adaptation of the salt-water balance in animals. PMID- 2547670 TI - Reproducible high yields of rat islets of Langerhans. AB - Isolation of islets of Langerhans from the pancreas by action of collagenase, a major breakthrough for physiological studies in vitro, has long appeared empirical, and the results were sometimes unpredictable. Isolation yields (number of islets obtained per pancreas) and their reproducibility, purification from exocrine remnants and vitality of the islets obtained, can improve owing to precise techniques, adapted to the architecture of collagen in the pancreas. We have tested four isolation-purification techniques in the rat pancreas. The best results were obtained by combining intra-ductal collagenase injection, with complete but moderate distension of the gland, avoiding leakage, multistep digestion (in situ and then in vitro), followed by purification on a discontinuous bovine serum albumin (BSA) gradient. Average yields were 670 +/- 40 islets per pancreas (range 570-800), versus 170 +/- 9 (range 40-350) with the technique used initially. The use of BSA discontinuous gradient improved the purification yield: 90-96% of islets obtained were concentrated in the 26/29% BSA interface. Furthermore, this technique shortened the duration of purification step: 1 hr (centrifugation gradient) vs 3 hrs (handpicked). It was verified that islets were morphologically free of exocrine tissue. Islet structure was well preserved either in conventional histology or insulin and glucagon immunoperoxidase staining. Islet vitality, as assessed by trypan blue exclusion test, was 100% of freshly isolated islets, and 89% after 24 hrs culture. Insulin secretory responses to a given stimulus were stronger than in the case of islets isolated by former techniques: 10-12 times the basal release (vs 5 times) with clear dose-response proportionality.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547671 TI - Microinjection of endogenous and exogenous proteins into primary cultures of rat hepatocytes and the degradation of the injected proteins. AB - 1. A method to microinject proteins into cells through packaging proteins to erythrocyte ghosts (erythrocyte-mediated microinjection) was modified partially in order to apply the method to primary cultures of rat hepatocytes. 2. Degradation of the microinjected proteins was examined employing the improved method. The mean half-life of the injected endogenous liver protein was 20 hr. The data suggested that the injected proteins are degraded through both lysosomal and non-lysosomal proteolytic pathways probably depending on their structure. 3. The present method to microinject exogenous proteins into primary cultures of rat hepatocytes can be employed usefully for the investigations of protein metabolism in liver. PMID- 2547672 TI - Effect of 2,3-butanedione on human myeloperoxidase. AB - 1. Myeloperoxidase is inhibited by various diketones that are recognized arginine reagents. 2. Although arginine residues in the enzyme were modified in both the light and the dark, enzyme inactivation occurred only in the presence of light. 3. Under conditions where diketones caused inactivation of myeloperoxidase, spectral studies indicated marked damage to the haem residues of the enzyme. 4. It was concluded that diketones serve simply as photosensitizers of visible light induced inactivation of myeloperoxidase. 5. Studies on other haemoproteins indicated the great ease with which the presence of diketones sensitized haem residues for photodestruction. PMID- 2547673 TI - Calmodulin-dependent phosphatase preferentially dephosphorylates a 28 kDa protein in human platelets. AB - 1. Human platelets contain a calmodulin-dependent phosphatase (calcineurin) that has many properties similar to those of bovine brain calmodulin-dependent phosphatase. 2. The activity of calcineurin phosphatase accounts for a small fraction of the total phosphatase activity in human platelets. 3. Labeling of human platelets with 32P yielded many phosphoproteins. 4. Incubation of a lysate of the 32P-labeled platelets with bovine brain calmodulin-dependent phosphatase led to preferential dephosphorylation of a 28 kDa protein (P28), a minor component of platelet proteins. 5. P28 is one of several proteins that were rapidly labeled upon stimulation of platelets with thrombin. 6. Even though the enzyme is known to catalyze the dephosphorylation of many substrates in vitro, its apparent preference for P28 suggests that its activity is highly selective. PMID- 2547674 TI - Catecholestrogens inhibit proliferation and DNA synthesis of porcine granulosa cells in vitro: comparison with estradiol, 5 alpha-dihydrotestosterone, gonadotropins and catecholamines. AB - Studies were conducted to assess the role of catecholestrogens on ovarian follicular growth using cultured porcine granulosa cells. Effects of the catecholestrogens, 2-hydroxyestradiol (2-OH-E2) and 2-methoxyestradiol (2-MeO-E2) were compared to those of estradiol (E2). Treatment with saturating concentrations of 2-OH-E2 caused a significantly greater decrease in cell numbers measured after 2 days of treatment than E2 treatment. The inhibitory effect of 2 OH-E2 was time and concentration dependent, not associated with a change in the viability of cells, and was partially reversible. The potency of 2-MeO-E2 to inhibit cell numbers was similar to or greater than that of 2-OH-E2. 2-MeO-E2 had a greater inhibitory effect on DNA synthesis, as measured by [3H]thymidine incorporation into trichloroacetic acid-precipitable macromolecules, than 2-OH-E2 or E2 in the absence or presence of insulin, epidermal growth factor or platelet derived growth factor. Concurrent treatment with epinephrine significantly enhanced the inhibitory effect of 2-OH-E2 on granulosa cell DNA synthesis. Collectively, these studies indicate that catecholestrogens are more potent inhibitors of granulosa cell replication than E2 and 5 alpha-dihydrotestosterone (DHT), and that catecholamines may modulate the antimitotic activity of 2-OH-E2. These results support the hypothesis that catecholestrogens play a role in proliferation of granulosa cells during growth of ovarian follicles. PMID- 2547675 TI - Angiotensin II-stimulated cortisol secretion is mediated by a hormone-sensitive phospholipase C in bovine adrenal fasciculata/reticularis cells. AB - Conditions have been established for the incorporation of [3H]inositol ([3H]Ins) into the phosphoinositides of cultured bovine adrenal zona fasciculata/reticularis (ZFR) cells. Stimulation of these prelabelled cells with angiotensin II (10(-11)-10(-7) M AII) resulted in the dose-dependent (max. 16 fold at 10(-7) M AII), time-dependent formation of water-soluble radiolabelled products which show the same chemical and chromatographic properties as [3H]InsP, [3H]InsP2 and [3H]InsP3 standards. The results of the time-course studies of the changes in these products are consistent with the view that AII rapidly (less than 15 s) induces the activation of a polyphosphoinositide-specific phospholipase C. The action of this phospholipase on the polyphosphoinositides is sustained throughout 15 min of stimulation. The dose dependency of this response correlates closely with cortisol output and is reduced (to 52%, P less than 0.00005), but not abolished, in the absence of extracellular Ca2+. To our knowledge these results are the first clear demonstration that AII stimulates a polyphosphoinositide-specific phospholipase C in bovine ZFR cells. PMID- 2547676 TI - Identification of a cAMP-responsive region in thyroglobulin gene promoter. AB - The DNA sequences involved in transcription control by a cAMP-dependent mechanism have been localized in the thyroglobulin gene promoter region by a functional assay. The proximal 5'-flanking sequences from the bovine thyroglobulin gene were linked to the bacterial chloramphenicol acetyl-transferase gene. Transient expression of this reporter gene was studied in dog thyrocytes in primary culture in the presence, or absence, of cAMP stimulation. Deletion analysis showed that the cAMP-responsive region is contained within the first 250 base-pairs of the promoter, and suggests that it could correspond to a sequence conserved between species. These DNA sequences do not bear significant homology with cAMP responsive elements (CRE) described previously. By contrast, some similarities were found with the fat-specific element (FSE2) of genes under cAMP control in adipocytes and with DNA elements mediating cAMP-dependent regulation of expression of two different genes in the lower eukaryote Dictyostelium discoideum. This suggests that control of Tg gene transcription by cAMP could involve a mechanism different from the one mediated by a classical CRE. PMID- 2547677 TI - Adventitious viral agents in biological products. AB - The objectives of tests for extraneous agents will be discussed in the light of quality requirements for biological products published over the last twenty years, current developments of novel production methods and products, and past and current virological findings. PMID- 2547678 TI - Transfection and transformation of human and animal cells by recombinant DNA vectors: issues relevant to use of these substrates for the production of biologicals. AB - This paper reviews some of the issues relevant to the effect of oncogene or recombinant DNAs in both in vivo and in vitro models. Many studies of directly injected DNAs alone, with mediators of DNA uptake, or as the initiator in a multi stage tumor progression model, showed that the DNAs were only rarely (if at all) tumorigenic. Conclusions from these and other in vitro experiments were that single oncogenes transfected into human cells did not generally convert those cells to a malignant phenotype, suggesting that additional genetic insult(s) or other factors were needed. These data, in concert with other observations and standard methods for product purification, imply that recombinant DNAs or oncogenes in cell lines pose little or no risk in the production of biologicals. PMID- 2547679 TI - Rat pup isolation distress and the brain benzodiazepine receptor. AB - Pharmacologic studies have demonstrated that benzodiazepines can modulate the ultrasonic vocalizations (USV) associated with social separation of rat pups. In this study, in vivo receptor autoradiography was used to determine if brain benzodiazepine receptors were functionally less available to bind an exogenous ligand during social separation. The labeled benzodiazepine receptor antagonist. 3H-RO 15-1788, was given to 10-day-old rat pups with varying schedules of social separation. In initial studies with homogenized and solubilized tissue, we found a 30% reduction in binding to cortex when pups were separated for 25 min beginning 5 min prior to tracer injection. In subsequent autoradiographic studies with this same separation schedule, the binding of 3H-RO 15-1788 was examined in 21 brain regions. Again binding was decreased in neocortex (frontal, motor, and somatosensory). In addition, we found significantly decreased binding in hippocampus, dentate gyrus, and superior and inferior colliculi. These same regions showed no alteration of in vitro binding of 3H-RO 15-1788. Therefore, these decreases in in vivo binding do not reflect changes in receptor number. The interpretation of decreased in vivo binding and implications of these results for defining the neural substrates of separation behavior are discussed. PMID- 2547680 TI - Enalapril does not alter renal function in normotensive, normoalbuminuric, hyperfiltering type 1 (insulin-dependent) diabetic children. AB - Using a prospective randomised double-blind crossover design, the effect of the angiotensin converting enzyme inhibitor enalapril compared to a placebo was studied in 18 normotensive, normoalbuminuric Type 1 (insulin-dependent) diabetic children. Each patient had a high normal or clearly elevated glomerular filtration rate (145 ml.min-1.1.73 m2 or higher) in the 6 months prior to the study. Enalapril, 0.5 mg.kg-1.day-1, was given for 4 weeks followed by placebo for 4 weeks, or vice versa. At the end of each period, glomerular filtration rate, renal plasma flow, blood pressure, plasma renin activity, and converting enzyme activity were determined. Enalapril caused significant reduction (p = less than 0.001) in blood pressure and converting enzyme activity and a rise in plasma renin activity. A slight but not significant rise in glomerular filtration rate and renal plasma flow without change in filtration fraction was observed. These data suggest that the renin angiotensin system is not involved in the glomerular hyperfiltration of Type 1 diabetes, and can be interpreted as showing no evidence for the presence of intraglomerular hypertension in these patients. PMID- 2547681 TI - Impaired insulin-like growth factor I-mediated stimulation of glucose incorporation into glycogen in vivo in the ob/ob mouse. AB - The ability of insulin to modulate glucose metabolism is impaired in insulin resistant ob/ob mice. It has been shown that insulin-like growth factor I stimulates the uptake and metabolism of glucose in muscle through the insulin like growth factor receptor not the insulin receptor. Thus, we have compared the abilities of insulin-like growth factor I and insulin to stimulate the in vivo incorporation of [14C]-glucose into glycogen in the diaphragm of ob/ob mice and their lean littermates. The animals used in these studies were 12-14 weeks old and the serum insulin levels of the ob/ob mice were 16-fold higher than in their lean littermates. There were no differences in the serum levels of glucose or insulin-like growth factor I. Both insulin and insulin-like growth factor I stimulate the incorporation of [14C]-glucose into glycogen in lean mice. Significant stimulation occurs at doses as low as 1 micrograms/kg of either peptide. The effective doses of insulin and insulin-like growth factor I are quite similar, which indicates that the effect of insulin-like growth factor I is mediated by the insulin-like growth factor receptor and not the insulin receptor. In contrast, greater than 100 micrograms/kg of insulin-like growth factor I is required to stimulate [14C]-glucose incorporation into glycogen in the diaphragm of ob/ob mice. Thus, ob/ob mice are resistant to the action of both insulin and insulin-like growth factor I.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547683 TI - Preferential expression of cellular retinoic acid binding protein in a subpopulation of neural cells in the developing mouse embryo. AB - The cellular retinoic acid binding protein is thought to be involved in the retinoic-acid-mediated signal transduction pathway. We have isolated the mouse cellular retinoic acid binding protein cDNA from an embryonal-carcinoma-derived cell line by using differential cDNA cloning strategies. In situ hybridization on sections of mouse embryos of various developmental stages indicated that the cellular retinoic acid binding protein gene, which we localized on mouse chromosome 9, is preferentially expressed in a subpopulation of neurectodermal cells. This restricted expression pattern suggests an important role for cellular retinoic acid binding protein in murine neurogenesis. PMID- 2547684 TI - [Extragonadal tumors. Considerations on extragonadal germinal tumors and a description of 1 case]. AB - Germinal tumours compose about 1-2% of all human neoplasms. The most common histotypes appear to be seminoma and teratocarcinoma. These are the most frequent neoplasms in 15 to 35 years old males. Up until 15 years ago, the prognosis of these tumours was almost always poor. New combined chemo-radiotherapy approaches permit prolonged survival or a cure in most cases. We describe the case of a patient who benefitted from these new therapeutic protocols. A review of these new treatments is also presented. PMID- 2547682 TI - Islet cell and other organ-specific autoantibodies in all children developing type 1 (insulin-dependent) diabetes mellitus in Sweden during one year and in matched control children. AB - The majority (about 90%) of children developing Type 1 (insulin-dependent) diabetes mellitus do not have a first-degree relative with the disease. Nearly all (389/405, 96%) children (0-14 years) in Sweden, who developed diabetes during one year, were therefore studied to compare islet cell, thyroid peroxidase, thyroglobulin, and gastric H+, K+-ATPase antibodies with 321 age, sex, and geographically matched, but non-related, control children. Islet cell (cytoplasmic) antibodies were found in 81% (316/389) of the patients and in 3% (9/321) of the control children (p less than 0.001). The median islet cell antibody levels were 70 (range 3-8200) Juvenile Diabetes Foundation (JDF) Units in the islet cell antibody positive patients, and 27 (range 17-1200) JDF Units in the control children (NS). Autoantibodies against thyroid peroxidase (8%), thyroglobulin (6%), and gastric H+, K+-ATPase (3%) were all increased in the patients compared with the control children, being 2% (p less than 0.001), 2% (p less than 0.01), and 0.3% (p less than 0.01), respectively. During an observation time of 20-34 months, two of the nine islet cell antibody positive control children developed Type 1 diabetes, after 8 and 25 months respectively, while the others remained healthy and became islet cell antibody negative. None of the islet cell antibody negative control children developed diabetes during the same time of observation. This first investigation of an unselected population of diabetic children and matched control children shows: that islet cell antibodies are strongly associated with newly diagnosed childhood diabetes, that other autoantibodies are more frequent among diabetic children than control children, and that the frequency of islet cell antibodies in the background population of children is higher than previously documented, and could also be transient, underlining that factors additional to islet cell antibodies are necessary for the later development of Type 1 diabetes. PMID- 2547685 TI - [Neoplasm of the liver and spleen. A clinical case]. PMID- 2547686 TI - Wire-guided brush cytology: a new endoscopic method for diagnosis of bile duct cancer. PMID- 2547687 TI - Malignant tumors of the bile ducts: diagnosis by biopsy during endoscopic cannulation. PMID- 2547688 TI - Esophageal miliary granular cell tumor. PMID- 2547689 TI - Synthesis of Rhodobacter sphaeroides cytochrome c2 in Escherichia coli. AB - The cytochrome c2 structural gene, cycA, from Rhodobacter sphaeroides was expressed in Escherichia coli. CycA-specific mRNA was detected in E. coli both under aerobic and anaerobic conditions with trimethylamine-N-oxide as electron acceptor. However mature holocytochrome c2 was only detected in anaerobically grown cells. The mature form of cytochrome c2 (Mr = 12,500) was secreted into the periplasm of E. coli suggesting that the signal polypeptide was processed. The cytochrome c2 synthesized in E. coli exhibited absorbance maxima in the reduced form at 550 nm (alpha-band) and 521 nm (beta-band) and contained covalently attached haem c. The results indicate that a foreign c-type cytochrome can be secreted and assembled in E. coli under anaerobic conditions. PMID- 2547690 TI - pEG plasmid involved in styrene degradation: molecular dimorphism and integration of a segment into the chromosome. AB - In the Pseudomonas fluorescens strain ST the ability to utilize styrene as the sole carbon source is due to the presence of a plasmid, pEG. In the present report we show that pEG contains two inverted repeat sequences and we present evidence indicating that the catabolic genes are localized in these repeats. The region separating the inverted repeats can assume alternative orientations. In the chromosome of strain ST, a 3 kbp region is homologous to sequences present at one end of the plasmid repeats. This region is present in one copy in the chromosome and could be a specific site for integration of the plasmid. We suggest that this sequence, which is present twice in the pEG plasmid and once in the chromosome, might be a transposon-like element. PMID- 2547691 TI - Identification and characterization of a new gene of Escherichia coli K-12 involved in outer membrane permeability. AB - Using a genetic selection for mutations which allow large maltodextrins to cross the outer membrane of Escherichia coli in the absence of the LamB maltoporin, we have obtained and characterized two mutations that define a new locus of E. coli. We have designated this locus imp for increased membrane permeability. Mapping studies show that the imp gene resides at approximately 1.2 min on the E. coli chromosome. The mutations alter the permeability of the outer membrane resulting in increased sensitivity to detergents, antibiotics and dyes. The mutations are nonreverting and codominant. Genetic analysis of the mutations suggest that the imp gene is an essential gene. We describe a general cloning strategy that can be used to clone both dominant and recessive alleles. Using this technique, we have cloned the wild-type and mutant imp alleles onto a low copy number plasmid. PMID- 2547692 TI - Recombination between homologies in direct and inverse orientation in the chromosome of Salmonella: intervals which are nonpermissive for inversion formation. AB - Sequences placed in inverse order at particular chromosome sites (permissive) recombine to generate an inversion; the same sequences, placed at other sites (nonpermissive) interact recombinationally but do not form the expected inversion recombinants. We have investigated the events that occur between sequences at nonpermissive sites. Genetically marked lac operons in inverse order were placed at nonpermissive sites in a single chromosome and Lac+ recombinants were selected. No inversions were formed. The Lac+ recombinants recovered include double-recombinant types in which information appears to have undergone a nonreciprocal information exchange; one mutant copy is repaired with no alteration of the other copy. Recombination within the lac operon is stimulated more than 100-fold by the presence of extensive homology (antenna sequences) outside of the region for which recombination is selected. Sequences placed in direct order at the ends of the same noninvertible chromosome segment recombine to form all the expected recombinant types including those in which a reciprocal exchange has generated a duplication. All the detected recombinant types can be accounted for by recombination between sister chromosomes. These results are discussed in terms of two alternative models. One explanation of the failure to detect inversion of some intervals is that particular inversions are lethal, despite the fact that no essential sequences are disrupted. Another explanation is that chromosome topology prevents sequences at nonpermissive sites in a single chromosome from engaging in the direct interaction required for inversion formation, but allows the sister strand exchanges that can generate the recombinant observed. PMID- 2547693 TI - Mitotic and meiotic gene conversion of Ty elements and other insertions in Saccharomyces cerevisiae. AB - We examined meiotic and mitotic gene conversion events involved in deletion of Ty elements and other insertions from the genome of the yeast Saccharomyces cerevisiae. We found that Ty elements and one other insertion were deleted by mitotic gene conversion less frequently than point mutations at the same loci. One non-Ty insertion similar in size to Ty, however, did not show this bias. Mitotic conversion events deleting insertions were more frequently associated with crossing over than those deleting point mutations. In meiosis, conversion events duplicating the element were more common than those that deleted the element for one of the loci (HIS4) examined. PMID- 2547694 TI - The cloning of the Bar region and the B breakpoint in Drosophila melanogaster: evidence for a transposon-induced rearrangement. AB - We have cloned the B breakpoint in Drosophila melanogaster using DNA from a P-M induced revertant of B, which has a P element inserted at the B breakpoint. The analysis of the B DNA reveals that there is a transposable element, B104, right at the breakpoint. This suggests that this element may have been involved in the generation of the B breakpoint and the associated tandem duplication. One possible mechanism to generate the B duplication is a recombination event between two B104 elements, one at 16A1 and the other at 16A7. DNA sequencing data of the junctions of the B104 element support this model. Four partial revertants of B are the result of insertions of transposable elements very close to the B breakpoint. This supports the hypothesis that the breakpoint is the cause of the B mutation. The clones from B were used to isolate wild-type clones from 16A1, the location of the Bar gene. Four rearrangement breakpoints associated with various Bar mutations map within a 37-kb region, suggesting that the Bar gene is very large. PMID- 2547696 TI - [Mutants of Escherichia coli K-12 with altered excision efficiency of transposons Tn5 and Tn9]. AB - HFETn5, HFETn9 and LFETn9 mutants of Escherichia coli K-12 have been isolated. The frequency of Tn5 precise excision from the chromosomal lac operon is increased 3-660-fold in nine HFETn5 mutants. The majority of these mutations have no influence on the efficiency of precise excision of transposon Tn9, though hfeTn5-04 and hfeTn5-06 mutations decrease excision efficiency 2-13-fold. The Tn9 transposon is excised in HFETn9 mutant about 20-fold more efficiently than in the wild type strain. This mutation does not stimulate excision of Tn5 and Tn10. LfeTn9 mutation decreases excision frequency of Tn9 11-17-fold, but has no effect on Tn5 excision and increases that of Tn10 about 20-fold. The differences in genetic control and mechanisms of excision of the transposons with long and short inverted repeats are discussed. PMID- 2547695 TI - [Deletion-insertion mapping of the region non-essential for functioning of the beta-subunit of Escherichia coli RNA polymerase]. AB - A plasmid has been constructed containing the gene of beta-subunit of RNA polymerase of Escherichia coli under control of the PR promoter of bacteriophage lambda. PR promoter may be induced by heating up to 42 degrees C. In frame insertions of different sequences between 989 and 990 or 1010 and 1011 codons of the rpoB gene do not inactivate the beta-subunit function. Deletions in the region of 1011-1027 codons result in inactivation of beta-subunit. We localized antigene determinant of monoclonal anti-beta-antibodies which do not inactivate RNA polymerase in vitro. The borders of non-essential region of beta-subunit were accurately determined. PMID- 2547697 TI - [Restriction of two-replicon shuttle Escherichia coli-Streptomyces plasmids in Streptomyces lividans strain 66]. AB - The shuttle Escherichia coli - Streptomyces plasmids were used to transform S. lividans 66. Plasmid DNAs isolated from this strain transform it 10-1000-fold more efficiently than DNAs from E. coli. Rare transformant cured from most restricted plasmid is more efficient recipient of plasmid DNA from E. coli and has the property of R +/- M+ mutant. Restriction in S. lividans 66 correlates with the appearance in DNA from E. coli of the sites susceptible to Scg2I restriction endonuclease. The latter was isolated earlier from recombinant strain Rcg2, a hybrid between S. griseus Kr. 15 and S. coelicolor A3(2). Scg2I possesses the recognition sequence CCTAGG, like EcoRII, MvaI and Eco dcm methylase. The DNA resistant to Scg2I cleavage retained this ability after in vitro modification by EcoRII methylase. So, the resistance of DNA to Scg2I cleavage is not connected with methylation at 4th and 5th position of second cytosine in the recognition sequence. Neither restriction of plasmid DNA in S. lividans 66 is dependent on dcm modification in E. coli, though its dependence on dam modification is not excluded. It is assumed that the restriction in S. lividans 66 is specified by endonuclease analogous to Scg2I. PMID- 2547698 TI - [Characterization of multiple changes of antibiotic resistance characters in Streptomyces coelicolor A3(2)]. AB - Among mutants of Streptomyces coelicolor A3(2) studied which were sensitive to chloramphenicol (Cmls), strains sensitive to a number of antibiotics (ristomycin, tetracycline, polymyxin, lincomycin) amount of 46%. Antibiotic-sensitive mutants are capable to form different classes of resistant revertants with frequency varying from 10(-2) to 10(-6) in independent strains. Ristomycin-sensitive clones (Rims) have been found to occur with high frequency in Cmls strains and Cmlr revertants. Mutations mediating the Rims phenotype are mapped in a locus linked to the gene for resistance to chloramphenicol. The results obtained are discussed, in accordance with the notion about possible role of cml mutation in induction of secondary mutational changes in the genome of S. coelicolor A3(2). PMID- 2547699 TI - [Methyl-cytosine specific restriction in Escherichia coli K-12]. AB - Experiments on transformation of Escherichia coli K-12 cells by plasmids carrying RM systems with different recognition sites containing 5-methylcytosine have shown that the gene mcrB determines the function of restriction. The data obtained made it possible to believe that E. coli possesses no restriction system recognizing specifically cytosine methylated in position 4. PMID- 2547700 TI - [Toxicologic characteristics and hygienic standardization of the levels of sodium hydrocarbonate and potassium carbonate in the air of the work area]. AB - Toxicity and hazards of sodium hydrocarbonate (SH) and potassium carbonate (PC) were experimentally assessed. Both substances caused impairments of electrolytic equilibrium, protein metabolism, changes of functional and biochemical indicators of the cardiovascular system, etc. LD50 of SH for white rats constituted 9940 +/- 350 mg/kg, for mice 3360 +/- 210 mg/kg and of PC 2980 +/- 142 and 2570 +/- 142 mg/kg, respectively. The authors failed to define mean lethal concentrations of both substances. The following threshold values Limac were established at 74 mg/m3 for SH and at 54 mg/m3 for PC. Limch was respectively 14.9 and 4.7 mg/m3. MAC for SH was established at 5 mg/m3 and for PC at 2 mg/m3. PMID- 2547701 TI - Fibre and enteral nutrition. PMID- 2547702 TI - [Endothelial precipitates and cytomegalovirus infection]. AB - In patients with the typical features of intraocular cytomegalovirus infection endothelial precipitates were identified that had a characteristic morphology, which included subtle, non-pigmented linear deposits with reticular arrangement that can spread diffusely over the corneal endothelium without preference for the inferior quadrants. The appearance of these precipitates was comparable to heterocromic cyclitis. Specular microscopy identified lymphocytes and macrophages on the endothelial layer. The present observations are based on five case reports of necrotizing retinitis, typical for intraocular cytomegalovirus infection, in connection with the atypical endothelial precipitates. In four of the five patients reported, HIV-infection was confirmed. In one patient with serologically confirmed cytomegalovirus infection (however without HIV-infection), a diagnostic work-up of the aqueous humor was carried out. Local production of specific antibodies against cytomegalovirus was identified in the aqueous humor. Topical treatment with antiviral and/or steroidal drugs was unsuccessful. The antiviral drug gancyclovir was applied systemically in two patients, but there was no impact on the corneal precipitates described. No intravitreal therapy was given. If these endothelial changes can be regarded as pathognomonic for cytomegalovirus infection, thorough slit lamp examination may be important for diagnosis of the presence of or the reactivation of cytomegalovirus infection in the eye. So far, no correlation has been found between the time course of corneal/endothelial precipitation and necrotizing retinitis. PMID- 2547703 TI - [Retinitis in AIDS patients: diagnosis, follow-up and treatment]. AB - Of 147 patients with AIDS (Walter Reed 3-6), 28 showed signs of retinal infection. Toxoplasmic retinochoroiditis had developed in 5 eyes of 4 patients, whereas cytomegalovirus (CMV) retinitis occurred in 40 eyes of 24 patients. All patients with toxoplasmosis complained of visual symptoms at the first visit, but only 60% of patients with CMV retinitis had ocular symptoms at this time. The important problems involved in making a timely and correct diagnosis (early CMV retinitis vs cotton wool spots vs toxoplasmic retinitis; significance of laboratory data) are presented and discussed. The course of CMV retinitis is sometimes fast and devastating and requires immediate treatment to prevent blindness in patients who are in otherwise still fair general health. Eighteen patients (28 eyes) were treated by intravenous ganciclovir (DHPG) for 1-9 months. Initial therapy (10 mg/kg body wt., 2-4 weeks) led to regression of fundus lesions in all eyes. Under maintenance treatment (5 mg/kg BW body wt., 5 times a week), 14 eyes still demonstrated significant regression or cicatrization of the lesions, 9 eyes showed little progress and 5 eyes moderate recovery. However, 8 of 12 untreated eyes became legally blind before the patient died. None of the 23 treated eyes with useful initial visual acuity (greater than or equal to 0.2) lost visual function. Only in three cases did the drug have to be stopped because of serious side effects (severe leukopenia, pancytopenia, psychosis). Toxoplasmic retinochoroiditis healed in all affected eyes after specific treatment (pyrimethamine, sulfamethoxydiazine, clindamycin, and spiramycin in double or triple combination).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547704 TI - Effect of D-tryptophan-6-luteinizing hormone-releasing hormone on the tumoral growth and plasma sex steroid levels in cirrhotic patients with hepatocellular carcinoma. AB - Certain evidence suggests androgen dependence of hepatocellular carcinoma in cirrhotic patients. Consequently, it was postulated that antiandrogen therapy might be effective in the treatment of hepatocellular carcinoma. D-Tryptophan-6 luteinizing hormone-releasing hormone is a potent agonist analog of luteinizing hormone-releasing hormone which, when chronically administered, inhibits the pituitary gonadal axis and testicular androgen secretion in man. We studied the effects of D-tryptophan-6-luteinizing hormone-releasing hormone on tumoral growth in 17 male cirrhotic patients with hepatocellular carcinoma. After 3 to 6 months of therapy, no tumoral response was observed. Furthermore, measurements of plasma levels of testosterone, dihydrotestosterone, androstenedione, estradiol, estrone and sex hormone-binding globulin were performed before and 3 months after initiation of the antiandrogenic treatment. Before treatment, hypoandrogenism and hyperestrogenism were present; D-tryptophan-6-luteinizing hormone-releasing hormone induced a fall in plasma testosterone and dihydrotestosterone levels. Only a moderate decrease in estradiol and no modification of plasma estrone and sex hormone-binding globulin were found, indicating that the hyperestrogenemia of cirrhotic patients could be attributed to an increase in peripheral aromatization of androgens of adrenal origin. The inability of D-tryptophan-6-luteinizing hormone-releasing hormone to reduce the growth of hepatocellular carcinoma is not totally in disagreement with the concept of androgen dependence of hepatocellular carcinoma since D-tryptophan-6-luteinizing hormone-releasing hormone does not inhibit the production of androgens of adrenal origin. PMID- 2547706 TI - [Validity of immunohistology and in situ hybridization in the differential diagnosis of cytomegalovirus pneumonia and idiopathic interstitial pneumonia after allogenic bone marrow transplantation]. AB - Interstitial pneumonia (IP) is currently the most frequent and severe complication of allogeneic bone marrow transplantation (BMT). Post-BMT-IP is due partially to infection by cytomegalovirus (CMV-IP) and partially to idiopathic induction (IIP). Because of the different therapeutic consequences it is important to distinguish between these two kinds of pneumonia. Therefore, the validity of immunohistochemistry (IHC) and in situ hybridization (ISH) for the differential diagnosis of CMV-IP and IIP was studied. The investigations were performed using postmortem tissue samples (lung) of 23 patients decreased after allogeneic BMT. In 22 of the 23 patients the primary cause of death was IP (CMV 13x, idiopathic 7x, pneumocystis 1x, toxoplasmosis 1x). One patient died from thrombopenic cerebral bleeding. All 3 CMV-detecting systems tested disclosed certain advantages and disadvantages. The sensitivity of routine histology was 12/13 (92.3%), of IHC 13/13 (100%), and of ISH 12/13 (92.3%). The specificity corresponded to the order: ISH greater than IHC greater than histology. The application of ISH and IHC did not change significantly the routine histologic classification of pneumonias into CMV-IP and IIP. The validity of the three procedures used for detecting CMV in patients after allogeneic BMT is discussed in detail. PMID- 2547705 TI - Subclinical neurological and neuropsychological effect of infection with HIV. AB - Thirty one homosexual men with antibody to human immunodeficiency virus (HIV) but without major neurological complaints were assessed in a cross sectional study of neurological and neuropsychological function. Eleven patients had AIDS, 10 had persistent generalised lymphadenopathy (PGL), and 10 had HIV infection without PGL (called "well"). Thirteen age matched homosexual men without antibody to HIV acted as controls. Significant abnormalities were found in six on clinical neurological examination, in eight on nerve conduction studies, in six on electroencephalography, in six on neuropsychological assessment, and in eight on computed tomography of the head. Eighteen patients (nine with AIDS, four with PGL, and five "well") performed abnormally in at least one section of the assessment. The study highlights the incidence of nervous system dysfunction in HIV infection even in people who do not have AIDS. Prospective evaluation using electrophysiological and imaging techniques is necessary to assess the natural history of such manifestations and the effect of antiviral treatment. PMID- 2547708 TI - Primitive neuroectodermal tumours of the central nervous system--an electron microscopic & immunohistochemical study. AB - A series of 75 poorly differentiated neoplasms of the central nervous system viz., medullo-blastoma, cerebral neuroblastoma, pineoblastoma and ependymoblastoma were studied by light microscopy (LM), electron microscopy (EM) and immunohistochemistry (IH). Although the predominant cells constituting these tumours appeared to be undifferentiated cells by LM and EM, many others showed evidence of differentiation into glial, neuronal or ependymal cell lines by EM and IH. It is therefore, concluded that all these neoplasms are best considered as primitive neuroectodermal tumours and these may be classified as such in neuro oncology. They may be subclassified further on the basis of differentiation into one or more cell lines. PMID- 2547707 TI - Glycogen content & structure & some enzymes of glycogen metabolism in human foetal organs. AB - The glycogen content, and its structure and the enzymes involved in glycogenolysis in human foetal organs were studied at different periods of gestation. Of all the tissues studied glycogen content was found to be the highest in cardiac muscle. Very little glycogen was present in the foetal liver at 9-12 wk of gestation, this increased progressively to nearly 2 per cent at 24 wk. Glycogen content of placenta was lower than that of skeletal muscle and liver. The level of glycogen in adipose tissue, placenta and cerebrum was not high enough to play any role in glucose homeostasis of the foetus. Human foetal liver and skeletal muscle glycogen showed the normal branched structure while the liver glycogen was found to be unusually stable. Glycogen phosphorylase activity in the foetal liver and muscle was found to be low, i.e., about a fifth and a fourth of adult liver and muscle activity respectively. The stability of foetal liver glycogen and phosphorolytic activity in the liver and muscle indicate negligible glycogenolysis during foetal development. Glucose-6-phosphatase activity in foetal liver was undetectable below 12 wk of gestation, the activity increasing progressively up to 24 wk. PMID- 2547709 TI - Caffeine may potentiate adrenocortical stress responses in hypertension-prone men. AB - The effect of caffeine on blood cortisol levels and blood pressures was examined during rest and in response to a challenging psychomotor task in men with a low versus high risk of essential hypertension. Thirty-four healthy men ages 21-35 years were selected such that 17 were at high risk for hypertension (positive parental history and screening blood pressures of 135/85-155/95 mm Hg) and 17 were at low risk (negative parental history and no pressures above 132/84 mm Hg). Testing consisted of quiet rest (20 minutes); oral placebo (grapefruit juice) or caffeine administration (3.3 mg/kg in grapefruit juice); rest during a postdrug absorption period (40 minutes); work on an unsignalled simple reaction time task (15 minutes); and quiet rest (20 minutes). Blood pressures were recorded at 2 minute intervals, and blood samples were withdrawn via an indwelling catheter at the end of the baseline, drug absorption, task, and recovery periods. The combination of task plus caffeine produced the highest blood pressures in men at risk for hypertension. Cortisol levels were found to be sustained during rest in members of the high risk group after they had consumed caffeine, whereas members of the low risk group showed a modest decline. The high risk subjects also showed a significant rise in cortisol during (+3.7 micrograms/dl) and after (+4.0 micrograms/dl) work on the reaction time task after caffeine consumption.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547710 TI - Renal nerve contribution to NaCl-exacerbated hypertension in spontaneously hypertensive rats. AB - Previous studies demonstrate that bilateral renal denervation enhances urinary sodium excretion and delays the onset of hypertension in young (7-week-old) spontaneously hypertensive rats (SHR) maintained on ordinary laboratory chow. We interpret these data as suggesting that increased renal nerve activity in this model contributes to hypertension by causing excess sodium retention. More recent studies show that dietary NaCl supplementation increases blood pressure and peripheral sympathetic nervous system activity in NaCl-sensitive SHR (SHR-S). The present study tests the hypothesis that the renal nerves contribute to the rise in arterial pressure caused by dietary NaCl supplementation in this model. SHR-S were fed a high (8%) or basal (1%) NaCl diet beginning at age 7 weeks. Bilateral renal denervation was carried out 2 weeks after the initiation of the diets, at which time systolic blood pressure was significantly higher in the high (compared with the basal) NaCl group. Systolic blood pressure was reduced slightly less in denervated SHR-S on the high (compared with basal) NaCl diet during the following 5 weeks. Renal denervation performed 1 week before initiation of the diets attenuated the subsequent development of hypertension equally in both groups. Both renal denervation and the high NaCl diet increased alpha 2-adrenergic receptor numbers in the kidney; renal denervation caused an approximately equal increase in alpha 2-adrenergic receptor binding in SHR-S on high and basal NaCl diets. The high NaCl diet increased plasma noradrenaline concentration, and renal denervation lowered mean arterial pressure but did not decrease circulating catecholamines in either diet group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547711 TI - Lysophosphatides enhance superoxide responses of stimulated human neutrophils. AB - Human neutrophils which are pretreated with subtoxic concentrations of a variety of lysophosphatides (lysophosphatidylcholine, lysophosphatidylcholine oleoyl, lysophosphatidylcholine myrioyl, lysophosphatidylcholine stearoyl, lysophosphatidylcholine gamma-O-hexadecyl, lysophosphatidylinositol, and lysophosphatidylglycerol) act synergistically with neutrophil agonists phorbol myristate acetate, immune complexes, poly-L-histidine, phytohemagglutinin, and N formyl-methionyl-leucyl-phenyalanine to cause enhanced generation of superoxide (O2-). None of the lyso compounds by themselves caused generation of O2-. The lyso compounds strongly bound to the neutrophils and could not be washed away. All of the lyso compounds that collaborated with agonists to stimulate O2- generation were hemolytic for human red blood cells. On the other hand, lyso compounds that were nonhemolytic for red blood cells (lysophosphatidylcholine caproate, lysophosphatidylcholine decanoyl, lysophosphatidylethanolamine, lysophosphatidylserine) failed to collaborate with agonists to generate synergistic amounts of O2-. However, in the presence of cytochalasin B, both lysophosphatidylethanolamine and lysophosphatidylserine also markedly enhanced O2 generation induced by immune complexes. O2- generation was also very markedly enhanced when substimulatory amounts of arachidonic acid or eicosapentanoic acid were added to PMNs in the presence of a variety of agonists. On the other hand, neither phospholipase C, streptolysin S (highly hemolytic), phospholipase A2, phosphatidylcholine, nor phosphatidylcholine dipalmitoyl (all nonhemolytic) had the capacity to synergize with any of the agonists tested to generate enhanced amounts of O2-. The data suggest that in addition to long-chain fatty acids, only those lyso compounds that possess fatty acids with more than 10 carbons and that are also highly hemolytic can cause enhanced generation of O2- in stimulated PMNs. PMID- 2547712 TI - Acute inflammatory lung injury retards pulmonary particle clearance. AB - The deposition, retention, and clearance of inhaled cobalt oxide particles from the lungs of calves with acute inflammatory lung injury induced by parainfluenza 3 virus (PI-3) were examined. Acute pulmonary inflammation was induced by nebulization with 10(9) TCID50 of PI-3 virus on two successive days, and animals were subsequently exposed to an aerosol of particulate cobalt oxide (geometric mean diameter 0.54-0.65 microns) seven days post-virus infection (dpi). Pulmonary lesions at 7 dpi were typical of PI-3 pneumonitis and were characterized by patchy aveolitis and bronchiolitis with accumulations of neutrophils, macrophages, fibrin, and inflammatory debris. Calves were killed at 0, 7, and 21 days post-aerosol exposure (dpe) to evaluate particle clearance and retention by assay for cobalt in lung tissues, bronchoalveolar washings, and tracheobronchial lymph nodes. Control animals had a typically biphasic clearance pattern with rapid initial clearance of 50% of the initial lung burden (ILB) by 7 dpe followed by slower prolonged clearance. Clearance was significantly retarded (P less than 0.05) in calves with viral-induced acute inflammatory lung injury; 90% of the ILB was retained at 7 dpe. Essentially all particles recoverable by bronchoalveolar lavage were intracellular within pulmonary alveolar macrophages (PAM) in both experimental and control groups, but interstitial sequestration of particles within PAM was commonly observed only in the lungs of calves with viral pneumonitis. Pneumonic calves also exhibited retarded translocation of particles to regional lymph nodes. The results document impaired particulate clearance from acutely inflamed lungs, and implicate decreased mucociliary clearance and interstitial sequestration within PAM as the major contributing factors. These functional alterations would be expected to enhance the progression of virus induced acute pulmonary inflammatory injury. PMID- 2547714 TI - Ontogeny of inflammatory cell responsiveness: superoxide anion generation by phorbol ester-stimulated fetal, neonatal, and adult bovine neutrophils. AB - Newborn calves, like human infants, are uniquely susceptible to bacterial infections. Part of this increased susceptibility may be related to defects in newborn polymorphonuclear leukocyte (PMN) defensive functions. It remains unclear whether reported deficits in newborn PMN function represent maturational disorders or are manifestations of some form of perinatal suppression phenomenon. We therefore compared the ability of bovine newborn PMNs (less than 24 h old), newborn PMNs (7-10 days of age), fetal PMNs (210-220 days gestational age), and adult PMNs to generate superoxide anion (O2-) as an indicator of respiratory burst activity. Citrated blood was collected, and PMNs were isolated to greater than 95% purity and 98% viability. O2- generation was measured as the superoxide dismutase-inhibitable (10 micrograms/ml) reduction of ferricytochrome c (2 mg/ml) after activation of PMNs with phorbol myristate acetate (PMA, 2 micrograms/ml) to directly stimulate protein kinase C. The reaction kinetics were measured (37 degrees C, 550 nm) using a spectrophotometer and chart recorder for continuous monitoring. O2- generation was measured for 5 min after the initial lag period and the total nanomoles of O2- generated calculated using the extinction coefficient for ferricytochrome c. Newborn PMNs (N = 10) generated significantly less O2- (5.7 +/- 0.8 nmol O2-/10(6) cells/5 min, P less than 0.01) than did adult PMNs (N = 14) (9.6 +/- 2.1 nmol O2-/10(6) cells/5 min) or fetal PMNs (N = 4) (10.7 +/- 0.7 nmol O2-/10(6) cells/5 min). PMNs from 7- to 10-day-old calves (N = 9) generated almost identical amounts of O2- as newborn PMNs (5.7 +/- 1.6 nmol O2-/10(6) cells/5 min). There was no difference in measured lag time period between newborn and adult PMNs, but fetal PMNs had significantly reduced (P less than 0.01) mean lag time. The data indicated that bovine newborn PMNs have a decreased ability to generate O2- in response to PMA stimulation, which persists for at least 7-10 days, and that this functional decrement may be a manifestation of some form of perinatal PMN suppression phenomenon rather than a developmental abnormality since fetal PMNs produced O2- as well as adult PMNs. PMID- 2547715 TI - Metal content of neutrophil granules is altered in chronic inflammation. AB - The mass fraction of certain elements was measured in isolated granulocytes and isolated granulocyte granule fractions from patients with active inflammatory arthritides (N = 6) and healthy controls (N = 6). The patients had significantly increased amounts of Ca in the granulocytes, in the specific and light azurophil granules, but normal Ca amounts in the dense azurophil granules. Sr was below the detection limit in the granulocytes and granule fraction from controls, but it appeared in high concentrations in the granulocytes and all granule fractions from the patients. The patients had considerably increased granulocyte amounts of Mn but only slightly increased Mn concentrations in the specific granules. Mn was not detectable in azurophil granules from patients and controls. A prominent accumulation of Fe was seen in the granulocytes from the patients, together with an Fe accumulation in the specific granules. Fe was below the detection limit in azurophil granules from patients and controls. The patients had reduced granulocyte Zn and reduced amounts of Zn in the dense and light azurophil granules but normal Zn amounts in the specific granules. The results obtained indicate that the granulocyte accumulation of Ca, Sr, and Fe observed during chronic inflammation is associated with corresponding granule accumulation of these metals; the considerable Mn accumulation in granulocytes during inflammation is not localized in their granules; and the granule subpopulations differ in their capacity to store certain metals. PMID- 2547713 TI - Effect of piroxicam therapy on granulocyte function and granulocyte elastase concentration in peripheral blood and synovial fluid of rheumatoid arthritis patients. AB - The effect of piroxicam therapy (20 mg/day for 15 days) on various polymorphonuclear granulocyte (PMN) responses and on PMN elastase concentration was investigated in nine patients with active rheumatoid arthritis. Peripheral blood and synovial fluid samples were collected before starting therapy and 12 h after the last dose of the drug. All patients were evaluable for peripheral blood analysis and six for synovial fluid analysis. Piroxicam therapy had no effect on PMN random migration and phagocytosis, while it significantly reduced both FMLP induced aggregation and FMLP-induced chemotaxis. This seems mainly due to an effect on FMLP binding, as no differences were observed after therapy in PMA- and PHA-induced aggregation as well as in serum-induced chemotaxis. In contrast, a marked impairment of NBT test and PMA- and FMLP-induced superoxide anion (O2-) production was found after piroxicam therapy. This effect was as evident in peripheral blood as in synovial fluid PMN. Also, a significant reduction in synovial fluid PMN number and synovial fluid PMN elastase concentration (elastase alpha 1-proteinase complex) was found after treatment. It is concluded that piroxicam may act at different sites on various PMN responses. Its effect on O2- generation and PMN elastase concentration in synovial fluid may have an important role in reducing destruction of arthritic joint tissue. PMID- 2547716 TI - Effects of pyocyanine, a phenazine dye from Pseudomonas aeruginosa, on oxidative burst and bacterial killing in human neutrophils. AB - The effects of pyocyanine (phenazinium, 1-hydroxy-5-methyl-hydroxide, inner salt) on oxidative burst in human polymorphonuclear leukocytes were studied by several different approaches. In a cell- and enzyme-free system, pyocyanine oxidized NADPH. The reduced pyocyanine could be measured by its reaction with ferricytochrome c. It was shown by this assay that resting as well as phorbol myristate acetate- or zymosan-stimulated granulocytes reduced pyocyanine. The effect was independent of mitochondria, as cytoplasts were similarly active. Measurement of the hexose monophosphate shunt in intact granulocytes in the presence of pyocyanine indicated a concentration-dependent activation of the shunt without the generation of O2-, suggesting that pyocyanine oxidizes NADPH to NADP+ when it enters granulocytes. Intracellular NADPH in granulocytes was indeed lowered by almost 40% after incubation with pyocyanine. It is by this shuttling of reduction equivalents, leading to the partial depletion of NADPH, that pyocyanine affects the observed concentration-dependent partial inhibition of the phorbol myristate acetate- and zymosan-stimulated generation of O2-. A further consequence was that the intracellular killing of Staphylococcus aureus was also partially suppressed, particularly at higher loads of granulocytes with bacteria. Phagocytosis was not inhibited by pyocyanine concentrations as high as 500 microM. Pyocyanine did not affect the intracellular killing of Pseudomonas aeruginosa. The possible relevance of these findings to the course of mixed hospital infections in immunocompromised patients is discussed. PMID- 2547717 TI - Mapping of the human plasmin domain recognized by the unique plasmin receptor of group A streptococci. AB - A high-affinity surface receptor for human plasmin has been reported on certain group A streptococci. To map the region of the plasmin molecule that binds to the bacterial receptor, isolated domains of plasmin were tested for their ability to inhibit the binding of intact radiolabeled plasmin to receptor-positive bacteria. Complete inhibition of binding of labeled plasmin to bacteria by isolated heavy chains was achieved, but this inhibition was not as efficient on a molar basis when compared with that of unlabeled plasmin. By contrast, a conformationally altered form of native plasminogen was found to bind to bacteria and was as efficient a competitive inhibitor as intact plasmin was. The results of this study indicate that the selective binding of human plasmin to a group A streptococcus is dependent on structures present in the conformationally altered form of native plasminogen or plasmin that are not found on the native zymogen, the plasminogen with NH2-terminal glutamic acid. PMID- 2547718 TI - Invasion of HeLa 229 cells by virulent Bordetella pertussis. AB - Phase-dependent invasive behavior of Bordetella pertussis was demonstrated by recovery of viable organisms from gentamicin-treated HeLa cell monolayers and by transmission electron microscopy. Several mutants of B. pertussis with Tn5 or Tn5 lac inserted into various vir-regulated genes were evaluated for differences in their invasive abilities. Mutants lacking filamentous hemagglutinin, pertussis toxin, and two as yet uncharacterized vir-regulated products had levels of invasion significantly lower than that of the parent strain BP338. In contrast, invasion by mutants lacking adenylate cyclase toxin was significantly increased compared with that of wild-type B. pertussis. This increase in invasion was eliminated when concentrations of intracellular cyclic 3'-5' AMP were stimulated by treating HeLa cells with cholera toxin or forskolin. Entry of B. pertussis occurred through a microfilament-dependent phagocytic process, as evidenced by the marked reduction in uptake following treatment of HeLa cells with cytochalasin D. Invasion was inhibited with polyclonal anti-B. pertussis and anti filamentous hemagglutinin antisera. In addition, a monoclonal antibody against lipooligosaccharide A reduced uptake by 65.5%. The preservation of HeLa cell integrity and the limited replication of intracellular bacteria suggest that invasion may represent a means by which B. pertussis evades an active host immune response. PMID- 2547719 TI - Effects of gamma interferon on syntheses of DNA and proteins by Entamoeba histolytica. AB - To define the participation of cell-mediated immunity in resistance to amebic infection through the action of soluble mediators or lymphokines (LKs), including gamma interferon (IFN-gamma), we studied their effect on Entamoeba histolytica. Supernatants from cultures of lymphoid cells, which had been stimulated in vitro with concanavalin A and were rich in lymphokines (LRSNs), and recombinant IFN gamma were used. LRSN and recombinant IFN-gamma inhibited the growth of E. histolytica trophozoites in vitro. These LKs did not show a cytotoxic effect on the ameba, but they did inhibit rather significantly protein and DNA syntheses of the protozoa. Interestingly, LRSN incubated at 4 degrees C in the presence of trophozoites lost the ability to inhibit the replication of vesicular stomatitis virus. IFN-gamma inactivated at pH 2 had no effect on DNA synthesis by the ameba, thus suggesting that IFN-gamma is responsible for the observed inhibition of parasite growth. Furthermore, the IFN-gamma inhibitory effect was abolished by a monoclonal antibody specific for this LK. The results suggest that IFN-gamma may participate in protection against amebiasis infection through the activity of mediators released by lymphocytes during infection. PMID- 2547720 TI - Activity of polymorphonuclear leukocytes in the presence of sulfide. AB - Polymorphonuclear leukocytes (PMN) isolated from human blood were exposed to various levels of hydrogen sulfide. The effect on respiratory burst, myeloperoxidase activity, and capacity to phagocytose and kill bacteria were studied. A 1-h exposure of the PMN to 1 mM sulfide did not decrease their myeloperoxidase activity or their capacity to initiate a respiratory burst. Actually the products of the respiratory burst rapidly oxidized sulfide. The phagocytosis and killing of bacteria in the presence of 1 mM sulfide was only decreased to a minor extent. Myeloperoxidase in cell extract was, however, almost completely inhibited by 1 microM sulfide. These results indicate that hydrogen sulfide does not easily permeate PMN. PMN may be able to function in infected sites with high sulfide levels such as in the gingival pockets of periodontal disease. In the oxygenated areas of these sites the PMN may actually help in the detoxification of sulfide. PMID- 2547721 TI - Dynamics of dibutyryl cyclic AMP- and prostaglandin E2-mediated suppression of lipopolysaccharide-induced tumor necrosis factor alpha gene expression. AB - The regulation of lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNF) production by prostaglandin E2 (PGE2), forskolin, and dibutyryl cyclic AMP (cAMP) was examined at the cellular and molecular levels. The above three agents could suppress LPS (100 ng/ml)-stimulated TNF production by immunologically activated murine macrophages (M phi s) in a dose-dependent manner. The concomitant addition of PGE2, dibutyryl cAMP, or forskolin to LPS-challenged M phi s resulted in 50% inhibition of TNF production at 10(-7), 3 X 10(-6), and 3 X 10(-5) M, respectively. Interestingly, delaying the addition of PGE2 or dibutyryl cAMP by 1.5 h post-LPS stimulation was also effective in suppressing the production of TNF bioactivity, but only dibutyryl cAMP was effective when its addition was delayed by 3 h. Northern (RNA) blot analysis of mRNA isolated from LPS-challenged M phi s treated with PGE2 or dibutyryl cAMP corroborated the bioactivity data. The delayed addition of PGE2 or dibutyryl cAMP by 1.5 h post LPS stimulation resulted in a suppression of TNF mRNA accumulation by 50 to 70%. These data support the concept that LPS is a potent stimulus for M phi-derived TNF production and that this mediator is a very proximal signal in LPS-mediated disease states. Thus, therapeutic approaches that target the suppression of TNF in LPS-dependent disease states may be limited by the rapid expression of this mediator. PMID- 2547723 TI - Unusual precipitins in pathological human sera. AB - In conducting double diffusion gel precipitation tests, unexpected, strong precipitation reactions were noticed between a serum originating from a renal graft recipient and many pathological human sera. The highest frequency of positive reactions were produced by sera of patients with infectious mononucleosis (83%), chronic Epstein-Barr virus infections (72%), rheumatoid arthritis (57%), lepromatous leprosy (57%), and AIDS (44%). The precipitin in these pathological sera was identified as an antibody of IgM variety and the precipitinogen in the transplantation serum was shown to be a thermostable component with beta-globulin mobility. No explanation of the nature of reactions observed can be given at present. PMID- 2547722 TI - Monoclonal anti-idiotypic antibodies regulate the expression of virus-induced murine myocarditis. AB - A monoclonal anti-idiotypic antibody (anti-Id), produced by electrofusion and designated anti-Id88, was able to modulate expression of murine autoimmune myocarditis mediated by coxsackievirus B3 (CVB3). The anti-Id was characterized as an immunoglobulin G2b species possessing kappa light chains and was able to reduce expression of inflammatory myocarditis in anti-Id-pretreated mice challenged with CVB3. Anti-Id88 was able to stimulate specific cell-mediated immunity against anti-Id88, as well as CVB3, and exerted a suppressive effect on the proliferation of mixed spleen cell populations from virus-exposed mice. Anti Id stimulated an anti-anti-Id antibody 3 population able to bind antibody 2 F(ab')2 fragments or virus antigen in an indirect enzyme-linked immunosorbent assay. Western blot (immunoblot) analysis of anti-Id88 exhibited binding of syngeneic anti-Id antibody to idiotypes present on immunoglobulin G molecules from virus-immunized mice. PMID- 2547724 TI - Modulation of human peripheral blood eosinophil function by tumor necrosis factor alpha. AB - Eosinophils are important effectors in helminthic parasitic infection. Tumor necrosis factor (TNF-alpha) has been implicated as a mediator in the host response to parasitic infection and enhances eosinophil-mediated helminthotoxicity. We have examined the direct effects of recombinant human (rh) TNF on eosinophil functions of degranulation and oxidative metabolism. This report describes the minimal effects of rhTNF-alpha on eosinophil superoxide anion generation and enzyme secretion, which do not satisfactorily explain the observed increases in helminthotoxicity. In contrast to other cell types, eosinophils are unique in their differential responses to interleukin-1 beta and TNF. PMID- 2547725 TI - Prolonged exposure of neutrophils to saline or bronchoalveolar lavage fluid does not alter superoxide anion generation. AB - The role played by neutrophils (PMNs) in the genesis of lung injury in diverse clinical situations, such as bronchial asthma, idiopathic pulmonary fibrosis, and the adult respiratory distress syndrome, is an area of intensive investigation. Functional studies of PMNs, particularly those obtained from the alveoli by bronchoalveolar lavage, should shed light on their contribution to lung injury. However, it has not been demonstrated whether procedures used to harvest cells from the lung (bronchoalveolar lavage), particularly the potentially prolonged exposure to saline, commonly used to perform lavage, and other components of lavage fluid, can alter the functional characteristics of PMNs. In this report we demonstrate that a 2- to 3-hour exposure of neutrophils to saline from both humans and sheep in vitro does not alter the functional characteristics of PMNs as determined by superoxide anion generation after activation with phorbol myristate acetate (PMA; 6.96 +/- 0.44 vs. 7.60 +/- 0.32 nmol O2-/250,000 PMNs for control and saline-treated human cells, respectively, after a 45-min incubation with 10(-7) M PMA, and 4.73 +/- 0.30 vs. 4.50 +/- 0.42 nmol O2-/250,000 PMNs for control and saline-treated sheep cells). In a second series of experiments, we studied the effect of exposure of human PMNs to bronchoalveolar lavage fluid supernatants obtained from normal volunteers on superoxide anion generation by neutrophils.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547726 TI - Effects of cold exposure on cyclic AMP concentration in plasma, liver, and brown and white adipose tissues in cold-acclimated rats. AB - Effects of acute cold exposure on plasma energy substrates and tissue 3',5' adenosine monophosphate (cAMP) were analyzed in intact rats, to define an involvement of the nucleotide in nonshivering thermogenesis (NST) and resultant cold acclimation. After an acute cold exposure to -5 degrees C, the plasma glucose level increased gradually in warm-kept control rats (C) while it decreased significantly in cold-acclimated rats (CA). However, it was increased considerably by an extreme cold exposure to -15 degrees C in both C and CA. By contrast, plasma levels of free fatty acids (FFA) increased immediately after cold exposure and the release lasted during the period of exposure especially in C. The cold exposure also increased plasma cAMP concentration but no concomitant increase was found in the liver. In both brown (IBAT) and white (WAT) adipose tissues the nucleotide concentration showed a stepwise decrease. The observed correlation between lipolysis and plasma cAMP response after cold exposure suggests an involvement of the adenylate cyclase-cAMP system in NST via lipid metabolism, at least, in the early stages of cold acclimation. PMID- 2547727 TI - The male factor in the etiology of cervical cancer among sexually monogamous women. AB - To address the hypothesis that male sexual behavior may affect the etiology of invasive cervical cancer, a case-control study was undertaken in Panama, Costa Rica, Colombia and Mexico. The study enrolled husbands of those women with invasive cervical cancer and of those age-matched controls who reported only one lifetime sexual partner. A total of 204 case and 485 control husbands (78% and 72%, respectively, of identified husbands) were interviewed, clinically examined, and had penile swabs taken for papillomavirus assays. Risk increased significantly (p = 0.005) with the number of sexual partners reported by the husband (RR = 2.0 for 26+ vs. less than 6 partners). Low educational status of the husband was also an important predictor of risk, possibly indicating the role of unmeasured aspects of sexual behavior. Visits to prostitutes, circumcision status and sexually transmitted disease histories were not important predictors of risk, but evidence from clinical examination indicated that poor genital hygiene may be involved. Human papillomavirus (HPV) expression as defined by filter in situ hybridization was detected in 20-23% of subjects and, except in the small group with both HPV types 6/11 and 16/18, was not related to risk. This may reflect sampling problems in the male or the importance of host factors which enhance viral carcinogenicity in the female. PMID- 2547728 TI - Diverse responses to retinoid in morphological differentiation, tumorigenesis and N-myc expression in human neuroblastoma sublines. AB - We established the subline RT-BMV-C6 from the parent human neuroblastoma cell line RT-BM by a process that required repeated subculture of cells, which were prone to disaggregation. RT-BMV-C6 and the parent cloned line, RT-BM-1, had an identical marker chromosome, confirming that both lines were derived from a common progenitor. In the analysis of surface antigen expression, RT-BMV-C6 did not react with UJ-127-11, Leu7 or KP-NAC2 MAbs to which RT-BM-1 showed positive binding. The levels of both N-myc amplification and expression in RT-BMV-C6 were twice as high as the level obtained in RT-BM-1. Colony-forming efficiency in soft agar was 2.0 +/- 0.8% for RT-BMV-C6 and 3 times greater than that for RT-BM-1 (0.6 +/- 0.1%). When 100 x 10(6) cells of RT-BM-1 and RT-BMV-C6 were inoculated into nude mice, tumor incidence was significantly higher for RT-BMV-C6 (6/6; 100%) than for RT-BM-1 (0/6; 0%). Our data show that N-myc is closely related to tumorigenicity in NB. When RT-BM-1 and RT-BMV-C6 were co-cultured with a new synthetic retinoid, polyprenoic acid (E5166), and dibutyryl cyclic AMP, RT-BM-1 was induced to neuronal differentiation, defined by the formation of neuronal processes and expression of neurofilaments, whereas RT-BMV-C6 was not. However, when exposed to E5166, N-myc expression of RT-BMV-C6 was more strongly reduced than that of RT-BM-1, and colony formation of RT-BMV-C6 was significantly inhibited as compared to RT-BM-1. These findings suggest that the reduction of N myc expression might closely correlate with growth inhibition accompanying neuronal differentiation of neuroblastoma cells. PMID- 2547729 TI - The accelerating role of Abelson murine leukemia virus in murine plasmacytoma development: in vitro infection of spleen cells generates donor-type tumors after transfer to pristane-treated BALB/c mice. AB - The role of Abelson murine leukemia virus (A-MuLV) in the accelerated development of murine plasmacytomas (PCs) (Potter et al., 1973: Science, 132, 592-594) was studied in a new experimental system. Spleen cells from pristane-treated or untreated BALB/c mice carrying Robertsonian 6;15 fusion chromosomes were infected in vitro with helper-free A-MuLV overnight and subsequently transplanted into the peritoneal cavity of pristane-treated or untreated BALB/c mice. Donor-derived PCs developed in 4 out of 76 pristane-treated recipients [latent periods: 38-82 (mean 51) days] that had received spleen cells from pristane-treated donors, and also in 2 out of 41 pristane-treated recipients that had received untreated donor derived spleen cells (latent periods: 65 and 120 days). Three of the PCs in the former and both PCs in the latter group were tested for integration and expression of the v-abl gene, with positive results. This indicates that the spleen contains PC-precursor cells that can be activated by A-MuLV even before the impact of pristane. All 6 donor-origin PCs carried a translocation involving chromosome 15, band D2/3. Four of these corresponded to a typical 12;15 translocation, one was a variant 6;15 translocation and the 6th may represent a previously unidentified translocation between chromosome 15 and the lambda gene carrying chromosome 16. No PCs developed among 29 pristane-untreated recipients that had received pristane-treated donor-derived spleen cells. In addition to PCs, monocytic tumors developed in 37 (26%) of all recipients. Their development was independent of pristane treatment of recipients but was particularly frequent in those who had received spleen cells from pristane-treated donors. PMID- 2547730 TI - Molecular cloning and characterization of endogenous SV40 DNA from human HBL-100 cells. AB - The human HBL-100 cell line harbours SV40 DNA integrated in tandem at a unique site. The SV40 T-antigen expressed in these cells is defective in a function essential to the replication of the viral genome. The integrated SV40 sequences were molecularly cloned in a bacteriophage, and a subclone (plasmid pSVHBI) containing a complete SV40 DNA was isolated. As compared to SV40 wild-type strain 776, sequence analysis of pSVHBI early region revealed the presence of several DNA alterations. Among these, a point mutation at position 3199, predicting a change at amino-acid 540 of arginine to isoleucine, was shown by marker rescue to be responsible for the deficiency of T-antigen. This novel mutation further delimits one of the T-antigen domains involved in SV40 DNA replication. Transfection experiments demonstrated that the transforming activity of the SV40 genome from HBL-100 cells is still preserved. Moreover, several transformed human cell clones thus obtained could be permanently established in culture. PMID- 2547731 TI - From theory to practice: the planned treatment of drug users. Interview by S. Einstein. PMID- 2547732 TI - Herpes simplex virus type-1 strain influence on chorioretinal disease patterns following intracameral inoculation in Igh-1 disparate mice. AB - We have previously shown that the Igh-1 locus on chromosome 12 of the mouse influences contralateral disease patterns in the modified von-Szily model. Following intracameral injection with 1.5 X 10(4) PFU of HSV strain KOS (HSV KOS), 75% of BALB/cByJ (Igh-1a), 30% of C.AL-20 (Igh-1d) and 5% of C.B-17 (Igh 1b) mice develop contralateral chorioretinal necrosis. In contrast, Igh-1 congenic mice do not develop contralateral chorioretinal necrosis following anterior chamber inoculation of the same dose of an HSV-KOS mutant lacking surface glycoprotein-C (HSV-GC-KOS). Similarly, injection of wild type HSV strain mP (HSV-mP) into the anterior chamber of susceptible BALB/cByJ mice induces destructive contralateral chorioretinitis whereas injection of the same dose of the mutant HSV strain MP (HSV-MP) lacking surface glycoprotein-C does not induce a destructive contralateral chorioretinitis. In addition, higher doses of HSV-mP inoculum abrogate the Igh-1-associated contralateral chorioretinal protection seen in C.B-17 mice; protection is restored in C.B-17 mice at lower HSV-mP doses that still cause contralateral chorioretinitis in BALB/cByJ mice. These data demonstrate the influence of the viral isolate on the modified von-Szily model and specifically the requirement for the presence of glycoprotein-C on the surface of HSV for the development of contralateral destructive chorioretinitis. PMID- 2547733 TI - Comparative neuropharmacology of dihydroergotamine and sumatriptan (GR 43175). AB - The potency of dihydroergotamine (DHE) was determined at 13 neurotransmitter receptors using radioligand binding techniques. DHE is a potent agent (i.e. affinity value less than 100 nM) at 5-hydroxytryptamine1A (5-HT1A), 5-HT1C, 5 HT1D, 5-HT2, dopamine2, alpha1- and alpha2-adrenergic binding sites. DHE displays moderate affinity (i.e. affinity values = 100 - 1000 nM) for beta-adrenergic and dopamine1 sites and is completely inactive at 5-HT3, muscarinic and benzodiazepine receptors. These results were then compared to similar data on sumatriptan (formerly called GR 43175), a novel acute anti-migraine agent. The only pharmacological property shared by both agents is high affinity for 5-HT1D and 5-HT1A receptors. Therefore, these data suggest that 5-HT1D and/or 5-HT1A receptors may play an important role in the pathophysiology of migraine. PMID- 2547734 TI - Bioaccumulation and chemical modification of Tc by soil bacteria. AB - Bioaccumulation and chemical modification of pertechnetate (TcO4-) by aerobically and anaerobically grown soil bacteria and by pure cultures of sulfate-reducing bacteria (Desulfovibrio sp.) were studied to gain insight on the possible mechanisms by which bacteria can affect the solubility of Tc in soil. Aerobically grown bacteria had no apparent effect on TcO4-; they did not accumulate Tc nor modify its chemical form. Anaerobically grown bacteria exhibited high bioaccumulation and reduced TcO4-, enabling its association with organics of the growth medium. Reduction was a metabolic process and not merely the result of reducing conditions in the growth medium. Association of Tc with bacterial polysaccharides was observed only in cultures of anaerobic bacteria. Sulfate reducing bacteria efficiently removed Tc from solution and promoted its association with organics. Up to 70% of the total Tc in the growth medium was bioaccumulated and/or precipitated. The remaining Tc in soluble form was entirely associated with organics. Pertechnetate was not reduced by the same mechanism as dissimilatory sulfate reduction, but rather by some reducing agent released in the growth medium. A calculation of the amount of Tc that could be associated with the bacterial biomass present in soil demonstrates that high concentration ratios in cultures do not necessarily imply that bioaccumulation is an important mechanism for long-term retention of Tc in soil. PMID- 2547735 TI - Reaction mechanisms responsible for transformation of pertechnetate in photoautotrophic organisms. AB - Experimental data on the physiological effects of Tc on photoautotrophic and N2 fixing organisms all suggest a relation between their ability to generate strong reducing power and the incorporation of Tc. A series of biochemical experiments were undertaken to elucidate this problem. Isolated spinach chloroplasts, thylakoids and purified compounds of the photosynthetic electron transport chain were incubated with TcO4-. After illumination, the quantity of TcO4- transformed was measured with gel filtration chromatography. For part of the samples, the amount of extractable Tc(V) was determined. Isolated thylakoids showed reduction of TcO4- in the light, suggesting direct interference of TcO4- with the electron transport chain. Use of specific inhibitors and artificial electron carriers indicated that TcO4- withdraws electrons from ferredoxin. Competitive inhibition of TcO4- reduction by O2 and NADP+, as well as its capacity to function as a terminal acceptor in the diaphorase reaction with NADPH, indicates its interaction with the transport chain to be comparable to that of O2. In suspensions of thylakoids, TcO4- is mainly reduced into an extractable Tc(V) compound. Only part of the Tc fraction reduced by intact chloroplasts could, however, be extracted, whereas negligible quantities of unstable Tc(V) complexes were detected in intact plants. The stable complexes in vivo are supposed to originate through ligand exchange with strong complexing agents, such as thiol compounds. Disproportionation reactions of unstable Tc(V) compounds might result in complexes with Tc in lower oxidation states. PMID- 2547736 TI - Technetium-99 cycling in maple trees: characterization of changes in chemical form. AB - Prior field studies near an old radioactive waste disposal site at Oak Ridge, TN, indicated that following root uptake, metabolism by deciduous trees rendered 99Tc less biogeochemically mobile than expected, based on chemistry of the pertechnetate (TcO-4) anion. Subsequently, the form of technetium (Tc) in maple tree (Acer sp.) sap, leaves, wood and forest leaf litter was characterized using one or more of the following methods: dialysis, physical fractionation, chemical extraction, gel permeation chromatography, enzymatic extraction, or thin layer chromatography (TLC) on silica gel. Chromatography (Sephadex G-25) of TcO-4 incubated in vitro with tree sap showed it to behave similar to TcO-4 anion. When labeled wood and leaf tissues were processed using a tissue homogenizer, 15% and 40%, respectively, of the Tc was solubilized into phosphate buffer. Most (65% to 80%) of the solubilized Tc passing a 0.45-micron filter also passed through an ultrafiltration membrane with a nominal molecular weight cutoff of 10,000 atomic mass units (amu). A majority (72% to 80%) of the Tc in wood could be chemically removed by successive extractions with ethanol, water and weak mineral acid. These same extractants removed only 23% to 31% of the Tc from maple leaves or forest floor leaf litter. Most of the Tc in leaves and leaf litter was removed only by strongly alkaline reagents typically used to release structural polysaccharides (hemicelluloses) from plant tissues. Chromatography (Sephadex G 25) of the ethanol-water extract from wood and the alkaline extract from leaves demonstrated that Tc in these extracts was not principally TcO-4 but was complexed with molecules greater than 1000 amu. Incubations of leaf and wood homogenates with protease approximately doubled the amount of Tc released from contaminated tissues. Ultrafiltration of protease-solubilized Tc from leaves and wood showed that 40% and 93%, respectively, of the Tc was less than 10,000 amu. TLC of the less than 10,000 amu fraction indicated the presence of TcO-4 in wood but not in leaves. In the leaf, TcO-4 is converted to less soluble forms apparently associated with structural components of leaf cell walls. This conversion explains why 99Tc is not easily leached by rainfall from tree foliage and why 99Tc appears to accumulate in forest floor leaf litter layers at the Oak Ridge study site. PMID- 2547737 TI - The uptake of TcO-4 by plants: a mathematical description. AB - A model describing the uptake of TcO-4 by spinach plants was developed. The equation relates both plant and soil parameters (e.g., growth, metabolism, concentration of TcO-4 and composition of the growth medium) to the concentration of Tc in the shoot of the plant. As the soil solution is the medium from which plants obtain nutrients and non-nutrients, the modeling parameters have been obtained from uptake experiments using nutrient solutions (= simulated soil solutions) as the growth medium. Two important model assumptions are: 1) that an equilibrium exists between TcO-4 in the plant and the growth medium and 2) that the leaf TcO-4 metabolism is a pseudofirst order reaction occurring in a non constant volume. PMID- 2547738 TI - Technetium absorption and turnover in monogastric and polygastric animals. AB - Technetium (Tc) released into the environment can reach animals in various chemical forms: as pertechnetate (TcO-4) in drinking water or deposited on the surface of vegetables and forage plants, or as Tc bioincorporated into plants and associated with various plant constituents. In addition to being influenced by chemical speciation in the diet, absorption, metabolism, and retention of Tc in animals are modified by the treatment that the alimentary bolus undergoes during its passage through the gastrointestinal tract. This behavior differs markedly between polygastric and monogastric animals. We have, therefore, studied the fate of 99mTc given in the diet either as TcO-4 or bioincorporated into maize in rats (as an example of a monogastric animal) and in sheep (as an example of a polygastric animal). Urine and feces were collected and assayed for Tc activity by gamma spectrometry. Animals were sacrificed at different times after contamination, and the Tc content of tissues was determined. The pattern of absorption, excretion and, to a certain degree, of organ distribution and retention depended on animal species and species of Tc administered. Excretion was by feces and urine, and several metabolic components could be discerned. A component of very short half-time in urine suggests that newly absorbed Tc is more readily excreted than that already bound by tissues. The highest tissue concentrations were found in the thyroid. Retention of Tc was, however, most pronounced in bone and skin. Hair contains considerable amounts of Tc and may serve as a bioindicator of Tc contamination. PMID- 2547739 TI - Technetium metabolism by lactating goats. AB - The goat milk transfer coefficients of 99mTc, 95mTc and 99Tc, administered as pertechnetate (TcO4-), were 1.5 x 10(-4), 8.5 x 10(-4) and 1.1 x 10(-2) d L-1, respectively. The differences between the goat milk transfer coefficients of the three Tc isotopes administered as TcO4- were significant (p less than 0.05). The milk transfer coefficients of Tc were affected by chemical form and specific activity. Reduced 99mTc produced a transfer coefficient 17 times lower than that of 99TcO4- in the same goats. Reduction of pertechnetate in the rumen was evaluated to explain the low and variable transfer coefficients. PMID- 2547740 TI - Technetium metabolism in goats and swine. AB - The intestinal absorption of 99mTc given as pertechnetate (TcO4-) was studied in female goats and swine. In goats given TcO4- orally, only a small fraction of the given radioactivity was found in milk (x = 0.1% of dose) and urine (x = 1.2% of dose). These figures were not substantially changed by pre-treatment with I. Ninety percent was found in feces. In goats injected intraabomasally, secretion of 95mTc into milk increased fivefold and urinary excretion was doubled. In swine administered 95mTc intragastrically, urinary excretion was about 30% of the dose. Swine did not retain the radionuclide in the thyroid gland as long as the goats (T 1/2 20 h in swine versus 30 h in goats). Other organs containing substantial concentrations of Tc 200 h post-administration were livers and kidneys, but in swine the liver contained three times as much 95mTc as the thyroid or kidneys. The conclusion of these experiments would, however, be that the concentrations were insufficient to present a hazard to man from consumption of milk or meat. PMID- 2547741 TI - [Current aspects of radiotherapy of glomus jugulare tumors]. AB - Small glomus jugulare tumors can be operated on successfully. The intraoperative risk of bleeding may be reduced by preoperative irradiation or angiographic embolisation. Patients with advanced tumors (bone destruction, paralysis or cranial nerves and/or invasion of brain) are at high risk if they are operated on. In these cases radiotherapy is an effective alternative with a fairly low complication rate. Prerequisites for successful irradiation are assessment of treatment volume and treatment planning by CT, sophisticated stereotactic irradiation techniques, application of high energy photons and reliable immobilisation measures. Furthermore it is possible to deliver higher doses, leading to regression of the tumor and its symptoms. The technique is illustrated by two characteristic cases. Although irradiation alone can achieve tumor remission the long term prognosis remains doubtful. PMID- 2547742 TI - Classification of primary lung tumors in dogs: 210 cases (1975-1985). AB - Two hundred ten dogs that had primary lung tumors diagnosed between 1975 and 1985 were evaluated. The majority of the tumors were classified as adenocarcinoma (74.8%) and alveolar carcinoma (20%). The most common clinical signs of disease were cough (52%), dyspnea (23.8%), lethargy (18.1%), weight loss (12.4%), and tachypnea (4.8%). The clinical methods that were most successful in directly or indirectly leading to a diagnosis of primary lung tumor were thoracic radiography (77.1%) and cytologic examination of fine-needle aspirate specimens (24.8%). PMID- 2547743 TI - Microbial epimerization of 1-deoxynojirimycin to 1-deoxymannojirimycin. PMID- 2547744 TI - Efficacy of oxetanocin G against herpes simplex virus type 2 and murine cytomegalovirus infections in mice. PMID- 2547745 TI - Genetics of methicillin resistance in Staphylococcus aureus. PMID- 2547746 TI - Investigation of the structural properties of dihydrophenazines which contribute to their pro-oxidative interactions with human phagocytes. AB - Twenty-six dihydrophenazine compounds, including clofazimine, were investigated, at a fixed concentration of 1 mg/l, for their effects on spontaneous and stimulus activated generation of superoxide by human neutrophils in vitro. The synthetic chemotactic tripeptide N-formyl-L-methionyl-L-leucyl-L phenyl-alanine (FMLP) (0.1 microM) was used as a stimulant of membrane-associated oxidative metabolism. None of the agents tested influenced basal levels of superoxide generation by neutrophils. However sixteen of these compounds, all rimino phenazines, significantly increased the production of superoxide by FMLP-activated neutrophils. These pro-oxidative, priming interactions of the rimino phenazines with neutrophils were largely dependent on the nature of the alkylimino group at position 2 on the phenazine nucleus, and to a lesser extent on halogenation. Cycloalkylimino groups were generally less potent stimulants of superoxide generation by FMLP-activated neutrophils than clofazimine, and their pro oxidative properties were independent of mono- or dichlorination. However the halogen-free cycloalkylimino compound, B669, was an exceptionally potent pro oxidative agent. Chlorination was promotive to pro-oxidative activity in the case of dihydrophenazines with linear or branched alkylimino substituents. The pharmaco-therapeutic mechanisms of dihydrophenazines may be related to their pro oxidative interactions with phagocytes. PMID- 2547747 TI - Azithromycin (CP-62,993) in acute exacerbations of chronic bronchitis: an open clinical, microbiological and pharmacokinetic study. AB - A group of 21 patients admitted to hospital with acute purulent exacerbations of chronic bronchitis was treated for five days with a new oral macrolide azithromycin. They received one dose of 500 mg on the first day, followed by 250 mg once daily thereafter. Pre-treatment sputum cultures were typical for this district, but the cultures during and after treatment showed many Haemophilus influenzae infections persisting. Geometric mean MICs of azithromycin for these organisms rose from 1.23 mg/l (pre-treatment) to 4.87 mg/l, a week after the end of treatment. The mean MICs for erythromycin also rose four-fold. Streptococcus pneumoniae and Branhamella catarrhalis strains were effectively eradicated. Pharmacokinetic studies on serum and sputum from the first treatment day showed relatively low serum Cmax values, averaging 0.63 mg/l, but with considerable individual variation. However, peak sputum concentrations averaged 3.7 mg/l. Nevertheless, the clinical and microbiological results were not encouraging because of the failure to eradicate H. influenzae with the drug dosage used in this study. PMID- 2547748 TI - Synergy of clavulanic acid, sulbactam and tazobactam (YTR 830) with amoxycillin against fifty beta-lactamase-producing strains of Haemophilus influenzae. PMID- 2547749 TI - Effect of methotrexate alone and in combination with antifungal drugs on the growth of Candida albicans. AB - Methotrexate, at concentrations corresponding to drug levels found in vivo, caused a marked reduction in blastospore growth of Candida albicans strains in vitro, but did not affect germ tube elongation during the first 6 h after blastospores were shifted to conditions favouring hyphal formation. Combinations of methotrexate with amphotericin B, flucytosine and itraconazole showed no potentiation or antagonism of the antifungal effect of either component in tests with the blastospore form of C. albicans. In contrast, potentiation occurred when methotrexate-itraconazole combinations were tested for their effect on germ tube elongation of an azole-sensitive strain of C. albicans. This effect was not seen in tests with an azole-resistant strain. PMID- 2547750 TI - Norepinephrine induces lung vascular prostacyclin synthesis via alpha 1 adrenergic receptors. AB - Sympathetic nerve stimulation can cause pulmonary vasoconstriction related to norepinephrine (NE) release. Because of recent reports that NE caused prostacyclin (PGI2) release from systemic arteries, we wondered whether NE caused pulmonary vascular PGI2 release and whether a feedback mechanism existed whereby PGI2 modulated NE-induced vasoconstriction. NE-induced PGI2 synthesis in rat main pulmonary artery rings was larger than that induced by KCl, passive stretch, or a thromboxane analogue, was alpha-adrenergic receptor dependent, and was enhanced by endothelium removal. The NE-induced PGI2 synthesis was not tightly coupled to the magnitude of the pulmonary artery ring contractile response, and inhibition of NE-induced PGI2 production by cyclooxygenase blockade in either the pulmonary artery ring preparation or in isolated rat lungs perfused with a physiological solution did not augment the magnitude of the contractile response. We concluded that NE is a potent stimulus for PGI2 synthesis in the rat main pulmonary artery ring and in the rat lung, yet PGI2 is not important as a modulator of NE-induced vasoconstriction in the rat lung. PMID- 2547751 TI - Enzyme activities of FT and ST muscle fibers in heavy-resistance trained athletes. AB - Tissue samples were obtained from the vastus lateralis muscle of elite olympic weight and power lifters (OL/PL, n = 6), bodybuilders (BB, n = 7), and sedentary men (n = 7). Enzyme activities of citrate synthase (CS), lactate dehydrogenase (LD), 3-OH-acyl-CoA-dehydrogenase (HAD), and myokinase (MK) were assayed on freeze-dried dissected pools of slow-twitch (ST) and fast-twitch (FT) fiber fragments by fluorometric means. Histochemical analyses were carried out to assess fiber type composition and fiber area. CS and HAD activities were lower (P less than 0.05), and LD and MK were higher (P less than 0.05) in FT than ST fibers in the entire subject pool (n = 20). CS of FT fibers and HAD of ST fibers were lower in athletes (P less than 0.05-0.01) compared with nonathletes, whereas LD of both fiber types was higher (P less than 0.05-0.001) in athletes. CS activity of ST fibers and MK activity of FT fibers were higher (P less than 0.05) in BB compared with OL/PL. FT and ST fiber area was greater (P less than 0.05) in athletes than in nonathletes. BB displayed greater (P less than 0.05) fiber size than OL/PL. FT/ST area was greater (P less than 0.05) in OL/PL than BB. It is suggested that long-term heavy-resistance training results in specific metabolic adaptations of FT and ST fiber types. These changes appear to be influenced by the type of resistance training. PMID- 2547752 TI - Gas chromatographic determination of polychlorinated biphenyls (as Aroclor 1254) in serum: collaborative study. AB - A gas chromatographic-electron capture detection method for determining the concentration of polychlorinated biphenyls (PCBs) as Aroclor 1254 (AR 1254) in serum was evaluated through a 2-phase collaborative study. In Phase I, each collaborator's lot of Woelm silica gel (70-150 mesh) was evaluated for elution and recovery of AR 1254, which had been added in vitro at 25 ng/mL to a serum extract. In Phase II, each collaborator analyzed a series of bovine serum samples that contained the following: (1) in vitro-spiked AR 1254; (2) in vivo AR 1254 and 8 in vitro-spiked chlorinated hydrocarbons; (3) in vivo AR 1254 only; (4) 8 in vitro-spiked chlorinated hydrocarbons only; and (5) neither AR 1254 nor chlorinated hydrocarbons above the detection limit of the method. In Phase I, the average recovery of AR 1254 from silica gel for the 6 collaborators was 87.9 +/- 15.44% (mean +/- 1 SD; N = 18; range = 52.3-105.8%). In Phase II, the analysis of in vitro spikes of AR 1254 in serum at 8.58, 16.8, 41.8, and 84.3 ppb gave mean (means) interlaboratory recoveries of 89.0, 83.3, 79.4, and 76.9%, respectively, with within-laboratory (repeatability) relative standard deviations (RSDr) of 18.8, 20.5, 10.2, and 14.1%, respectively, and among-laboratory (reproducibility) relative standard deviations (RSDR) of 21.5, 21.1, 14.6, and 20.8%, respectively. The determination of in vivo AR 1254 in samples containing approximately 10, 25, 50, and 100 ng/mL of AR 1254 resulted in interlaboratory means of 10, 22, 39, and 79 ng/mL, respectively, with RSDr = 6.7, 9.7, 6.4, and 5.8%, respectively, and RSDR = 20.6, 16.0, 10.9, and 10.3%, respectively. The precision of the method for incurred AR 1254 showed a maximum RSDr of less than 10% and a maximum RSDR of less than 21% for a concentration range of 10-100 ng/mL. The accuracy of the method as demonstrated by the mean recovery of in vitro-spiked AR 1254 over a concentration range of 8.58-843 ng/mL was 82.2%. The method has been approved interim official first action. PMID- 2547753 TI - Heavy metal, pH, and total solid content of maple sap and syrup produced in eastern Canada. AB - Maple sap and syrups in eastern Canada were analyzed for pH, total solids, and the heavy metals Cu, Fe, Pb, and Zn. The levels of heavy metals found were within the range normally contained in food and water samples except for Pb. The concentration factor found in reducing sap to syrup did not reflect the same concentration change for the measured parameters. This indicates removal or conversion of heavy metals and organic acids with the sugar sands. There was no statistical difference among provinces with respect to the heavy metal, pH, and total solids content of sap. The only significant difference in syrup occurred with Cu and this appeared to be the result of the processing procedure. As the season progressed, the Cu, Pb, pH, and total solids content of the sap decreased while Zn increased and Fe showed little change. Syrups reflected a similar change. Statistical differences occurred in sap composition among sites within each province. PMID- 2547754 TI - Urea enzymatic dialysis procedure for determination of total dietary fiber. AB - A method that uses urea and enzymes for determination of total dietary fiber (TDF) in foods has been developed and compared with the AOAC enzymatic gravimetric method (43.A14-43.A20). In the evaluation, results for crude protein and ash contamination were higher by the AOAC method, particularly for samples that form gels during ethanol precipitation. The new urea enzymatic dialysis (UED) method quantitatively recovered, with less variation, more of the purified and semipurified dietary fiber products. TDF recoveries for carboxymethylcellulose and locust bean gum were 98% (SD 3.3) and 95% (SD 6.1) by the AOAC method and 99% (SD 1.0) and 100% (SD 0.6) by the UED method, respectively. The UED method was the more effective in removing starch. For kale samples, starch recovery was 3.5 and 0.2% from TDF residues obtained using the AOAC and UED methods, respectively. Differences were not significant among replicate values for determination of TDF in foods by the UED method (P greater than 0.01). Preliminary studies suggest that the new method can separately determine soluble and insoluble dietary fiber. The data indicate that the UED method is more precise and accurate than the AOAC method. PMID- 2547755 TI - Characterization of NADH kinase from Saccharomyces cerevisiae. AB - At least two enzymes that phosphorylate diphosphopyridine nucleotides were detected in Saccharomyces cerevisiae: NADH-specific kinase was localized exclusively in the mitochondria, and NAD+-specific kinase was distributed in the microsomal and cytosol fractions but not in the mitochondria. The identity of NAD+ kinase detected in the two fractions remains equivocal. NADH kinase was highly purified 1,041-fold from the mitochondrial fraction. The Km values for NADH and ATP were 105 microM and 2.1 mM, respectively. The relative molecular mass was estimated to be 160,000 by means of molecular sieve chromatography. From inactivation studies with SH inhibitors and protection by NADH, it was demonstrated that a cysteine residue is involved in the binding site of NADH. PMID- 2547756 TI - Purification and catalytic properties of a cross-linked complex between adrenodoxin reductase and adrenodoxin. AB - A stable covalent complex was prepared by cross-linking adrenodoxin reductase with adrenodoxin using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. The covalent complex was purified extensively until free components were removed completely. The major component of the complex had a molecular weight of 63 kDa, which corresponds to a 1:1 stoichiometric complex between adrenodoxin reductase and adrenodoxin. NADPH-cytochrome c reduction activity of the covalent complex was comparable to that of an equimolar mixture of adrenodoxin reductase and adrenodoxin (native complex), and the NADPH-ferricyanide reduction activity of the complex was equal to that of the native one. In contrast to the native complex, the covalent complex produced much less superoxide upon NADPH-oxidation, and the covalent complex was found to be more stable than the native complex, suggesting that the complex state is more favorable for catalysis. From these results, we conclude that the adrenodoxin molecule does not need to dissociate from the complex during electron transfer from NADPH to cytochrome c. PMID- 2547757 TI - Reactivity with ubiquinone of quinoprotein D-glucose dehydrogenase from Gluconobacter suboxydans. AB - D-Glucose dehydrogenase is a pyrroloquinoline quinone-dependent oxidoreductase linked to the respiratory chain of a wide variety of bacteria. There is a controversy as to whether the glucose dehydrogenase is linked to the respiratory chain via ubiquinone or cytochrome b. In this study, it was shown that the glucose dehydrogenase of Gluconobacter suboxydans has the ability to react directly with ubiquinone. The enzyme purified from the membranes of G. suboxydans was able to react with ubiquinone homologues such as ubiquinone-1, -2, or -6 in detergent solution. Furthermore, in order to demonstrate the reactivity of the enzyme with native ubiquinone, ubiquinone-10, in the native membranous environment, the dehydrogenase was reconstituted together with cytochrome o, the terminal oxidase of the respiratory chain, into a phospholipid bilayer containing ubiquinone-10. The proteoliposomes thus reconstituted exhibited a reasonable glucose oxidase activity, the electron transfer reaction of which was able to generate a membrane potential and a pH gradient. Thus, D-glucose dehydrogenase of G. suboxydans has been demonstrated to donate electrons directly to ubiquinone in the respiratory chain. PMID- 2547758 TI - Mapping of the active site tyrosine of eukaryotic DNA topoisomerase I. AB - DNA topoisomerase I from the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe was overproduced using the cloned genes. Extracts from cells overproducing DNA topoisomerase I were prepared and incubated with 32P labeled DNA. Alkali was used to trap the topoisomerase I-DNA covalent intermediate. Most of the DNA was digested with nuclease, and the resultant 32P labeled topoisomerase I was subjected to cleavage with cyanogen bromide or formic acid. From the molecular weights of the resultant labeled peptides and by comparison of the amino acid sequences derived from the cloned genes, we were able to deduce that the active site tyrosine of eukaryotic DNA topoisomerase I is very near the carboxyl terminus, at amino acid 771 for S. pombe and 727 for S. cerevisiae. Site-directed mutagenesis was used to change tyrosine 727 of S. cerevisiae topoisomerase I to a phenylalanine. The resulting mutant topoisomerase I protein lost all DNA relaxation activity and rendered cells resistant to the topoisomerase I inhibitor, camptothecin. The amino acid sequence of human topoisomerase I has significant similarity to the two yeast topoisomerase I sequences. Based on this similarity, we infer that tyrosine 723 is the active site tyrosine of human enzyme. PMID- 2547760 TI - Isolation and characterization of a yeast strain carrying a mutation in the mitochondrial promoter for COX2. AB - Subunit II of cytochrome c oxidase is a mitochondrially encoded protein required for cellular respiration. A respiration-deficient yeast strain has been isolated and shown genetically to carry a mutation in the subunit II structural gene COX2. The respiration-deficient strain produces no subunit II polypeptide and is missing two major transcripts of the subunit II gene. The mutation, first mapped by rho- recombinational rescue to the 5' end of the gene, has been localized precisely, by DNA sequencing, 58 nucleotides upstream of the COX2 ATG initiation codon. The mutant strain carries a single nucleotide change (A to T) relative to the wild type sequence. This mutation occurs in a sequence with substantial homology to a previously identified yeast mitochondrial promoter consensus sequence. These results provide the first in vivo evidence for the importance of the consensus sequence for promoter function. PMID- 2547759 TI - Purification and partial amino acid sequence of osteogenin, a protein initiating bone differentiation. AB - Osteogenin was purified from bovine bone matrix and its activity monitored by an in vivo bone induction assay. The purification method utilized extraction of the bone-inducing activity with 6 M urea, followed by chromatography on heparin Sepharose, hydroxyapatite, and Sephacryl S-200. Active fractions were further purified by preparative sodium dodecyl sulfate gel electrophoresis without reduction. Osteogenin activity was localized in a zone between 30 and 40 kDa. The amino acid sequences of a number of tryptic peptides of the gel-eluted material were determined. Reduction and alkylation of purified osteogenin in 7 M guanidine hydrochloride resulted in the total loss of biological activity. Sodium dodecyl sulfate gel electrophoresis under reducing conditions revealed a broad band with an apparent molecular mass of 22 kDa. PMID- 2547761 TI - Tocopheroxyl radical persistence and tocopherol consumption in liposomes and in vitamin E-enriched rat liver mitochondria and microsomes. AB - Substantial loading of rat liver mitochondrial and microsomal membranes with D alpha-tocopherol was achieved by dietary supplementation with no adverse effects of this loading being apparent, e.g. on treadmill exercise endurance. The tocopheroxyl radical was readily detected by ESR in the enriched microsomes and mitochondria. Continuous enzymatic oxidation with horseradish peroxidase and a hydrophilic phenol, to favor selective oxidation of tocopherol without the involvement of lipid peroxidation, allowed the tocopheroxyl radical to be observed for up to 1 h in liposomes of dioleoylphosphatidylcholine and for about 15 min in the subcellular membranes. Total alpha-tocopherol decreased throughout this period, but a significant residual fraction remained after all the ESR signal of tocopheroxyl had disappeared. Decay kinetics of the tocopheroxyl radical ESR signal produced by a burst of intense UV irradiation consisted of a rapid initial phase and a slower exponential decay. A more narrow and more persistent ESR signal, not yet chemically identified, was observed after the tocopheroxyl radical had disappeared under prolonged oxidation. Ascorbic acid prevented formation of the tocopheroxyl radical until the ascorbyl radical ESR signal had decayed, whereas uric acid, up to saturating concentration in phosphate buffer, had no effect. PMID- 2547762 TI - The ligand-binding conformation of Mr 46,000 mannose 6-phosphate-specific receptor. Acquisition of binding activity during in vitro synthesis. AB - Purified Mr 46,000 mannose 6-phosphate-specific receptor (MPR 46) lost its ligand binding activity after reductive alkylation and after enzymatic deglycosylation. Deglycosylated MPR 46 did not assemble to homodimers. Therefore, we investigated the role of N-glycosylation, intrasubunit disulfide bonds, and subunit assembly for the acquisition of ligand-binding activity during in vitro synthesis of MPR 46. Up to 21% of MPR 46 synthesized in a reticulocyte lysate supplemented with dog pancreas microsomes acquired ligand-binding activity provided that 1-5 mM glutathione was present during translation and during a chase following translation. Acquisition of ligand-binding activity after cotranslational membrane insertion and core glycosylation depended on formation of intrasubunit disulfide bonds and a conformational change. Formation of intrasubunit disulfide bonds was not sufficient for ligand-binding activity and is likely to precede the conformational change, which resulted in increased resistance toward trypsin, formation of highly antigenic epitopes, and association to dimers, concomitant with the acquisition of ligand-binding activity. PMID- 2547763 TI - Complementary DNA encoding core protein II of human mitochondrial cytochrome bc1 complex. Substantial diversity in deduced primary structure from its yeast counterpart. AB - Core protein II of mitochondrial cytochrome bc1 complex is the second largest nuclear-encoded subunit that is essential for the assembly of the functional complex. We have isolated, for the first time, a cDNA clone for a mammalian core protein II. Immunoscreening of a lambda gt11 rat liver cDNA library resulted in isolation of a cDNA encoding a partial sequence of rat mitochondrial core protein II, and this cloned cDNA was used as a probe for colony hybridization to screen a human fibroblast cDNA library. By comparison with the known NH2-terminal amino acid sequence of bovine core protein II, the amino acid sequence deduced from a cloned human cDNA was predicted to be composed of the part for the NH2-terminal extension of 14 amino acid residues and that for the mature polypeptide of 439 residues. The presequence is typical of precursors to mitochondrial proteins in being composed of a combination neutral and basic residues. The deduced sequence shows rather low homology with that of the corresponding subunit of yeast cytochrome bc1 complex; this finding is in contrast with the relatively high homology observed for the sequences of the redox center-carrying subunits of the same complex. On Northern blot analysis of human fibroblast poly(A)+ RNA, the cDNA hybridized to two mRNAs, a major species of 1700 nucleotides and a minor one of 2000 nucleotides. Southern blot hybridization of genomic DNA suggested that the human genome contains one or two genes for core protein II. PMID- 2547764 TI - Single strand DNA cleavage reaction of duplex DNA by Drosophila topoisomerase II. AB - A unique reaction for type II DNA topoisomerase is its cleavage of a pair of DNA strands in concert. We show however, that in a reaction mixture containing a molar excess of EDTA over Mg2+, or when Mg2+ is substituted by Ca2+, Mn2+, or Co2+, the enzyme cleaves only one rather than both strands. These results suggest that the divalent cations may play an important role in coordinating the two subunits of DNA topoisomerase II during the strand cleavage reaction. The single strand and the double strand cleavage reactions are similar in the following aspects: both require the addition of a protein denaturant, can be reversed by low temperature or high salt, and a topoisomerase II molecule is attached covalently to the 5' phosphoryl end of each broken DNA strand. Furthermore, the single strand cleavage sites share a similar sequence preference with double strand cleavage sites. There is, however, a strand bias for the single strand cleavage reaction. We show also that under single strand cleavage conditions, topoisomerase II still possesses a low level of double strand passage activity: it can introduce topological knots into both covalently closed or nicked DNA rings, and change the linking number of a plasmid DNA by steps of two. The implication of this observation on the sequential cleavage of the two strands of the DNA duplex during the normal DNA double strand passage process catalyzed by type II DNA topoisomerases is discussed. PMID- 2547765 TI - The identification and characterization of collagen receptors involved in HeLa cell-substratum adhesion. AB - Four proteins of molecular mass 102, 87, 45, and 38 kDa were isolated from plasma membrane preparations by affinity chromatography. The 102-, 87-, and 38-kDa proteins were shown to be collagen receptors involved in the adhesion of HeLa cells to a gelatin substratum. All four proteins were eluted by high salt from affinity columns made of either types I or IV collagen or type I gelatin. Generally, a total of six major proteins were found in the high salt eluates, although the relative amounts of each varied among experiments. Immunoprecipitation, immunoblotting, and limited peptide mapping indicated that the 102-kDa protein was most sensitive to proteolysis leading to the formation of proteins of molecular mass 58 and 54 kDa. Even in the presence of a mixture of protease inhibitors the 58-kDa fragment was usually the more abundant species. Lectin binding indicated that the 102-, 87-, and 38-kDa proteins contain carbohydrate. Phase-partitioning with Triton X-114 and the need to solubilize the proteins in Triton X-100 indicated that the 102-, 87-, 45-, and 38-kDa proteins have a hydrophobic domain. The 87-kDa protein partitioned exclusively with the detergent-rich phase, suggesting that it is the most hydrophobic. Cell surface labeling with 125I indicated that the four proteins have an extracellular domain. Four criteria were used to determine which of the four proteins are collagen receptors mediating cell-substrate adhesion: 1) during HeLa cell adhesion, proteins with Mr values similar to all four proteins or their peptide fragments were cross-linked to a gelatin substratum derivatized with a photoactivatable probe; 2) a pentapeptide containing the Arg-Gly-Asp cell recognition sequence eluted the same four proteins as those found by high salt elution of collagen affinity columns; 3) monospecific antibodies to the 102-, 87-, and 38-kDa proteins, but not the 45-kDa protein, inhibited the spreading of HeLa cells on a gelatin substratum; 4) monospecific antibodies to the 102-, 87-, and 38-kDa proteins, but not the 45-kDa protein, bound to culture dishes substituted for gelatin in mediating the spreading of HeLa cells. Taken together, the data suggest that the 102-, 87-, and 38-kDa proteins are collagen receptors involved in HeLa cell adhesion. Although the 45-kDa protein has two of the characteristics of a collagen receptor defined here, it does not fit the criteria for one involved in cell-substratum adhesion. PMID- 2547766 TI - Identification of two serine residues involved in agonist activation of the beta adrenergic receptor. AB - Pharmacophore mapping of the ligand binding domain of the beta-adrenergic receptor has revealed specific molecular interactions which are important for agonist and antagonist binding to the receptor. Previous site-directed mutagenesis experiments have demonstrated that the binding of amine agonists and antagonists to the receptor involves an interaction between the amine group of the ligand and the carboxylate side chain of Asp113 in the third hydrophobic domain of the receptor (Strader, C. D., Sigal, I. S., Candelore, M. R., Rands, E., Hill, W. S., and Dixon, R. A. F. (1988) J. Biol. Chem. 263, 10267-10271). We have now identified 2 serine residues, at positions 204 and 207 in the fifth hydrophobic domain of the beta-adrenergic receptor, which are critical for agonist binding and activation of the receptor. These serine residues are conserved with G-protein-coupled receptors which bind catecholamine agonists, but not with receptors whose endogenous ligands do not have the catechol moiety. Removal of the hydroxyl side chain from either Ser204 or Ser207 by substitution of the serine residue with an alanine attenuates the activity of catecholamine agonists at the receptor. The effects of these mutations on agonist activity are mimicked selectively by the removal of the catechol hydroxyl moieties from the aromatic ring of the agonist. The data suggest that the interaction of catecholamine agonists with the beta-adrenergic receptor involves two hydrogen bonds, one between the hydroxyl side chain of Ser204 and the meta-hydroxyl group of the ligand and a second between the hydroxyl side chain of Ser207 and the para hydroxyl group of the ligand. PMID- 2547768 TI - Agonist-induced regulation of inositol tetrakisphosphate isomers and inositol pentakisphosphate in adrenal glomerulosa cells. AB - In adrenal glomerulosa cells, angiotensin II (AII) rapidly stimulates the formation of inositol 1,4,5-trisphosphate (Ins-1,4,5-P3) and causes marked long term changes in the levels of highly phosphorylated inositols. Glomerulosa cells prelabeled with [3H]inositol for 48 h and exposed to AII for 10 min showed prominent increases in inositol 1,3,4,5-tetrakisphosphate (Ins-1,3,4,5-P4) and smaller increases in two additional tetrakisphosphates, Ins-1,3,4,6-P4 and another (Ins-3,4,5,6-P4) eluting in the position of Ins-3,4,5,6-P4 and its stereoisomer, Ins-1,4,5,6-P4, on anion exchange liquid chromatography. A concomitant decrease in InsP5 indicates that an increase in Ins-1,4,5,6-P4, the breakdown product of InsP5, is probably responsible for the initial rise in Ins 3,4,5,6-P4 during 10 min stimulation by AII. During prolonged stimulation by AII, Ins-1,3,4,5-P4 began to decline from its high, stimulated level after the first hour but the level of Ins-1,3,4,6-P4 remained elevated for several hours. There were also progressive increases in the levels of Ins-3,4,5,6-P4 and InsP5 during stimulation for up to 16 h with AII. Treatment of adrenal cells for 16 h with the cyclic AMP-mediated secretagogue, adrenocorticotropic hormone (ACTH), slightly increased basal levels of Ins-1,3,4,6-P4, Ins-3,4,5,6-P4, and InsP5, and enhanced the subsequent AII-stimulated increases in the two additional tetrakisphosphate isomers but not of inositol trisphosphates or Ins-1,3,4,5-P4. This change in the pattern of the higher inositol phosphate response to AII was manifested within 2 h after exposure to ACTH, and was mimicked by treatment with 8-bromo cyclic AMP or forskolin. Treatment with 50 microM cycloheximide abolished the ACTH-induced increases in inositol polyphosphate responses during AII stimulation, but had no effect on the responses of untreated cells to AII. The conversion of [3H]Ins 1,3,4-P3 to [3H]Ins-1,3,4,6-P4, a reaction linking the receptor-mediated InsP3 response to higher inositol phosphates, was enhanced in permeabilized cells that were pretreated for 16 h with either ACTH or AII. These results demonstrate that the reactions by which Ins-1,3,4,6-P4 and Ins-3,4,5,6-P4 are formed and converted to InsP5 are influenced by agonist-stimulated regulatory processes that include both calcium-dependent and cyclic AMP-dependent mechanisms of target cell activation. They also reveal changes consistent with agonist-induced conversion of InsP5 to its dephosphorylated metabolite, Ins-1,4,5,6-P4, during short-term stimulation by AII. PMID- 2547767 TI - Identification of receptors for platelet-activating factor in rat Kupffer cells. AB - Ligand binding studies demonstrated that isolated rat Kupffer cells possess high affinity binding sites for platelet-activating factor (1-O-alkyl-2-acetyl-sn glycero-3-phosphocholine, AGEPC). AGEPC binding reached saturation within 10 min at 25 degrees C and was reversible. A Scatchard analysis revealed a single class of AGEPC receptors numbering about 10,600 sites/cell and possessing a dissociation constant of 0.45 nM. Similar values for the dissociation constant for AGEPC (0.12 and 0.34 nM) were obtained independently by kinetic analysis of specific AGEPC binding. AGEPC binding was stereospecific and was inhibited by Zn2+ and AGEPC receptor antagonists including BN52021 and U66985. The AGEPC receptor was functionally active since it was shown to mediate arachidonic acid release and eicosanoid production in Kupffer cells, and these events were inhibited by AGEPC receptor antagonist BN52021. The receptor-mediated arachidonic acid release was extracellular calcium-dependent and was abolished by calcium channel blocker prenylamine and by [ethylenebis(oxyethylenenitrilo)]tetraacetic acid, indicating that calcium influx through a receptor-regulated calcium channel in the plasma membrane is involved in the AGEPC-induced arachidonic acid release. It is suggested that rat Kupffer cells have specific and functionally active AGEPC receptors which are involved in signaling mechanisms which govern the production of several other autacoid-type mediators in the liver. PMID- 2547769 TI - Binding, internalization, and intracellular processing of proteins interacting with recycling receptors. A kinetic analysis. AB - We measured time-dependent concentration changes of human interferon-alpha 2a (IFN) and human tumor necrosis factor-alpha (TNF) bound at the plasma membrane and internalized by human lung alveolar carcinoma A549 cells in the presence of excess free ligand. Concentration changes for these two ligands were substantially different. We modified our compartmental kinetic model encompassing receptor synthesis and receptor loss (Myers, A. C., Kovach, J. S., and Vuk Pavlovic, S. (1987) J. Biol. Chem. 262, 6494-6499) to include receptor recycling. We solved analytically the equations of three variants of the model of receptor recycling. All parameters (rate constants) were identifiable when the data sets consisted of time-resolved concentrations of IFN and TNF at the cell surface and internalized by cells. By least squares fitting we derived the best fit values for the first order rate constants for internalization of the ligand-receptor complex, receptor recycling, turnover of free receptors, elimination of the ligand from cells, and the rate of insertion of free receptors into the membrane. The best fit to data for interactions of cells with IFN was obtained without inclusion of the term for recycling of receptors to the membrane. The simplest model including receptor recycling was necessary and sufficient for the fit to the respective data for TNF. These results demonstrate that the contribution of receptor recycling to the metabolism of the ligand and the receptor can be quantitated by compartmental modeling. Receptor recycling does not contribute to the kinetics of Type I IFN receptor in A549 cells. In contrast, recycling contributes significantly to endocytosis mediated by the TNF receptor. PMID- 2547770 TI - Reconstitution of epidermal growth factor receptor transmodulation by platelet derived growth factor in Chinese hamster ovary cells. AB - Platelet-derived growth factor (PDGF) causes an acute decrease in the high affinity binding of epidermal growth factor (EGF) to cell surface receptors and an increase in the phosphorylation state of the EGF receptor at threonine654. The hypothesis that PDGF action to regulate the EGF receptor is mediated by the activation of protein kinase C and the subsequent phosphorylation of EGF receptor threonine654 was tested. The human receptors for PDGF and EGF were expressed in Chinese hamster ovary cells that lack expression of endogenous receptors for these growth factors. The heterologous regulation of the EGF receptor by PDGF was reconstituted in cells expressing [Thr654]EGF receptors or [Ala654]EGF receptors. PDGF action was also observed in phorbol ester down-regulated cells that lack detectable protein kinase C activity. Together these data indicate that neither protein kinase C nor the phosphorylation of EGF receptor threonine654 is required for the regulation of the apparent affinity of the EGF receptor by PDGF. PMID- 2547771 TI - The mechanism of cAMP-induced glucocorticoid receptor expression. Correlation to cellular glucocorticoid response. AB - The mechanism of regulation of glucocorticoid receptor (GR) expression by cAMP was investigated in rat hepatoma cells (HTC). Incubation of HTC cells with the cAMP-inducing agent, forskolin, caused a significant increase in the levels of both [3H]dexamethasone binding capacity and GR mRNA by about 2- to 2.4-fold within 4 h. Incubation of HTC cells with the cAMP analogue, 8-bromo-cAMP, also increased the GR mRNA level to a similar degree in a concentration-dependent manner. The increase in GR mRNA did not require ongoing translation or transcription. Determination of GR mRNA stability in 8-bromo-cAMP-induced cells showed that the message had a half-life of approximately 10 h, which is about 2.5 times longer than the GR mRNA half-life in nontreated cells (t1/2 = 4 h). These results indicate that the increased steady state level of GR mRNA induced by cAMP analogue is, at least in part, caused by increased GR mRNA stability. In both forskolin-pretreated and nontreated HTC cells, dexamethasone caused an approximately 70% down-regulation of GR protein levels. However, since forskolin induced the GR level 2- to 2.4-fold, the relative amount of GR protein remaining in cells treated with both forskolin and dexamethasone was about 2- to 2.4-fold higher compared to cells treated with dexamethasone alone. This increased GR level correlated well with the increase in inducibility of two glucocorticoid regulated genes, the endogenous tyrosine aminotransferase and the stably integrated mouse mammary tumor virus. These data suggest that relatively small changes in GR levels are reflected in parallel changes in cellular response to glucocorticoid hormones. This also implicates a limiting nature of the GR protein in determining the biological response. PMID- 2547772 TI - Immunological identification of 1,25-dihydroxyvitamin D3 receptors in human promyelocytic leukemic cells (HL-60) during homologous regulation. AB - Immunological techniques were utilized to detect 1,25-dihydroxyvitamin D3 (1,25 (OH)2D3) receptor levels and to characterize physical/chemical changes in receptors in human promyelocytic leukemic cells (HL-60) during continuous exposure to hormone. The monoclonal antibody (IVG8C11) raised against the porcine intestinal 1,25-(OH)2D3 receptor immunoprecipitated quantitatively 1,25-(OH)2D3 receptors in nuclear extracts from HL-60 cells. The highly enriched immunoprecipitated receptors were electrophoresed on sodium dodecyl sulfate polyacrylamide gels and transferred to polyvinylidene difluoride membranes, which were probed with 125I-labeled IVG8C11. The basal receptor from the cells treated with 1,25-(OH)2D3 for 2 h was detected as a single form at 53 kDa. Moreover, receptors were shown to be up-regulated at 12 h and down-regulated at 48 and 72 h in the continuous presence of hormone as evidenced by the ratio of density of the bands, 1.0 (2 h):4.2 (12 h):1.2 (48 h):0.9 (72 h), as measured by laser scanning densitometry. The up- and down-regulated receptors were also detected as single forms and had the same molecular mass as the basal receptor. Therefore, the data presented here strongly support the hypothesis of homologous regulation of 1,25 (OH)2D3 receptors in intact human target cells. PMID- 2547773 TI - Direct photolabeling of the cGMP-stimulated cyclic nucleotide phosphodiesterase. AB - cGMP-stimulated phosphodiesterase (PDE) has been directly photolabeled with [32P]cGMP using UV light. Sequence analysis of peptide fragments obtained from partial proteolysis or cyanogen bromide cleavage indicate that two different domains are labeled. One site, on a Mr = 36,000 chymotryptic fragment located near the COOH terminus, has characteristics consistent with it being close to or part of the catalytic site of the enzyme. This peptide contains a region of sequence that is highly conserved in all mammalian cyclic nucleotide PDEs and has been postulated to contain the catalytic domain of the enzyme. The other site, on a Mr = 28,000 cyanogen bromide cleavage fragment located near the middle of the molecule, probably makes up part of the allosteric site of the molecule. Labeling of the enzyme is concentration dependent and Scatchard analysis of labeling yields a biphasic plot with apparent half labeling concentrations of about 1 and 30 microM consistent with two types of sites being labeled. Limited proteolysis of the PDE by chymotrypsin yields five prominent fragments that separate by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) at Mr = 60,000, 57,000, 36,000, 21,000, and 17,000. Both the Mr = 60,000 and 57,000 apparently have blocked NH2 termini suggesting that the Mr = 57,000 fragment is a subfragment of the Mr = 60,000 fragment. Primary sequence analysis indicates that both the Mr = 21,000 and 17,000 fragments are subfragments of the Mr = 36,000 fragment. Autoradiographs of photolabeled then partially proteolyzed enzyme show labeled bands at Mr = 60,000, 57,000, and 36,000. Addition of 5 microM cAMP prior to photolabeling eliminates photolabeling of the Mr = 36,000 fragment but not the Mr = 60,000 or 57,000 fragments. The labeled site not blocked by cAMP is also contained in a Mr = 28,000 cyanogen bromide fragment of the enzyme that does not overlap with the Mr = 36,000 proteolytic fragment. Limited chymotryptic proteolysis also increases basal activity and eliminates cGMP stimulation of cAMP hydrolysis. The chymotryptic fragments can be separated by either ion exchange high performance liquid chromatography (HPLC) or solid-phase monoclonal antibody treatment. A solid-phase monoclonal antibody against the cGMP-stimulated PDE removes the Mr = 60,000 and 57,000 labeled fragments and any intact, unproteolyzed protein but does not remove the Mr = 36,000 fragment or the majority of activity. Ion exchange HPLC separates the fragments into three peaks (I, II, and III). Peaks I and II contain activity of approximately 40 and 100 units/mg, respectively. Peak II is the undigested or slightly nicked native enzyme.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2547774 TI - Mutation of the predicted ATP binding site inactivates both activities of isocitrate dehydrogenase kinase/phosphatase. AB - In Escherichia coli, the reversible phosphorylation of isocitrate dehydrogenase (IDH) is catalyzed by a bifunctional protein: IDH kinase/phosphatase. Although both IDH kinase and IDH phosphatase require ATP, the amino acid sequence of IDH kinase/phosphatase contains a single sequence that matches the consensus for ATP binding sites. A mutation that converted the "invariant" lysine (residue 336) of this consensus sequence to a methionine reduced the activities of both IDH kinase and IDH phosphatase by factors of greater than 500, to levels below the detection limits of the assays. The apparent elimination of both IDH kinase and IDH phosphatase by this mutation is consistent with the proposal that these activities share a common ATP binding site and that these reactions may occur at the same active site. Although conversion of Lys336 to a methionine eliminated detectable IDH kinase activity as measured in vitro, the mutant allele retained the ability to complement an aceK deletion mutation, restoring the ability of these cells to grow on minimal acetate medium. Complementation apparently resulted because the mutant protein retained sufficient activity to phosphorylate IDH in vivo. To determine whether the enzymatic assays performed in vitro had correctly reflected the activity of the mutant protein in vivo, we measured the rates at which mutant and wild-type cultures could incorporate [32P]inorganic phosphate into IDH. The wild-type culture achieved maximal incorporation in less than 3 min. In contrast, 32P incorporation was only barely detectable after 30 min in the mutant culture, indicating that the activity of the mutant protein is, indeed, greatly reduced in vivo. The ability of the mutant allele to complement an aceK null mutation thus suggests that IDH kinase/phosphatase levels in wild type cells are in great excess over what is required for steady-state growth on acetate medium. PMID- 2547775 TI - Localization of the membrane-associated CTP:phosphocholine cytidylyltransferase in Chinese hamster ovary cells with an altered membrane composition. AB - The subcellular localization of the membrane-associated CTP:phosphocholine cytidylyltransferase was determined in Chinese hamster ovary cells in which the phospholipid composition had been altered by growth in the presence of N methylethanolamine or treatment with phospholipase C. Cell homogenates were fractionated on Percoll density gradients, and marker enzyme activities were used to determine the location of the cellular membrane fractions. The peak of cytidylyltransferase activity occurred in the gradient at a density intermediate to that of the peaks of endoplasmic reticulum and plasma membrane markers. The profile of cytidylyltransferase activity most closely resembled that of the Golgi membrane marker; however, upon sucrose gradient centrifugation, the profile of the Golgi apparatus was very different from that of cytidylyltransferase. Differential centrifugation suggested a nuclear membrane association of the enzyme. Cytidylyltransferase was associated with a membrane fraction that sedimented when subjected to very low speed centrifugation (65 x g, 5 min). From Percoll gradient fractions, nuclei were identified by microscopy, and they migrated with cytidylyltransferase activity. The data are consistent with a localization of cytidylyltransferase in the nuclear membrane. PMID- 2547776 TI - Agonist regulation of beta-adrenergic receptor mRNA. Analysis in S49 mouse lymphoma mutants. AB - Agonist-promoted down-regulation of beta-adrenergic receptor mRNA was investigated in S49 mouse lymphoma variants with mutations in elements of hormone sensitive adenylate cyclase. In wild-type cells steady-state levels of beta adrenergic receptor mRNA were established by DNA-excess solution hybridization to be 1.72 +/- 0.08 (n = 8) amol/microgram total cellular RNA. Receptor mRNA levels declined 35-45% in response to stimulation by the beta-adrenergic agonist ( )isoproterenol or forskolin as described previously in DDT1 MF-2 cells (Hadcock, J. R., and Malbon, C. C. (1988) Proc. Natl. Acad. Sci. U. S. A. 85, 5021-5025). Agonist-promoted cAMP accumulation and down-regulation of receptor mRNA were analyzed in three variants with mutations in Gs alpha (H21a, unc, cyc-) and a single variant lacking cAMP-dependent protein kinase activity (kin-). H21a (Gs alpha coupled to receptor, but not to adenylate cyclase), unc (Gs alpha uncoupled from receptor), and cyc- (lacking Gs alpha) variants accumulated cAMP and down regulated beta AR mRNA in response to forskolin. In unc and cyc- cells isoproterenol failed to stimulate cAMP; accumulation and down-regulation of receptor mRNA was not observed. H21a cells, in contrast, displayed agonist promoted regulation of beta-adrenergic receptor mRNA but only basal levels of cAMP accumulation in response to isoproterenol. The kin- cells displayed cAMP accumulation in response to forskolin as well as to isoproterenol but no down regulation of receptor mRNA or receptor expression. Taken together these data demonstrate several features of agonist-promoted down-regulation of mRNA: (i) cAMP-dependent protein kinase activity is required for down-regulation of mRNA (kin-), although elevated cAMP accumulation is not (H21a); (ii) functional receptor-Gs coupling is required (H21a), and clones lacking Gs alpha (cyc-) or receptor Gs coupling (unc) lack the capacity to down-regulate mRNA in response to agonist; and (iii) in the presence of basal levels of cAMP and cAMP-dependent protein kinase activity, functional receptor-Gs coupling (H21a) to some other effector other than adenylate cyclase may be propagating the signal. PMID- 2547777 TI - Electron transfer through center o of the cytochrome b-c1 complex of yeast mitochondria involves subunit VII, the ubiquinone-binding protein. AB - The role of subunit VII, the ubiquinone-binding protein of the cytochrome b-c1 complex, in electron transfer reactions was investigated in yeast mitochondria. Preincubation of submitochondrial particles with specific antibody against subunit VII prior to addition of either succinate, NADH, or the reduced form of the decyl analogue of ubiquinol resulted in an approximately 40% increase in the extent of cytochrome c1 reduction compared with controls containing preimmune serum. Addition of antimycin, an inhibitor of center i, to submitochondrial particles resulted in a 21% decrease in the rate and a 36% decrease in the extent of cytochrome c1 reduction by succinate. Preincubation of submitochondrial particles with the antibody against subunit VII prior to addition of antimycin resulted in an increase in both the rate and extent of cytochrome c1 reduction to the levels observed in the control without inhibitor. The addition of myxothiazol (an inhibitor of center o), myxothiazol plus antimycin, or alkyl hydroxynaphthoquinone (an inhibitor analogue of ubiquinone) resulted in an almost complete inhibition in both the rate and extent of cytochrome c1 reduction; however, preincubation with the antibody against subunit VII prior to addition of these inhibitors resulted in a significant increase in cytochrome c1 reduction. These results confirm our previous report (Japa, S., Zhu, Q. S., and Beattie, D. S. (1987) J. Biol. Chem. 262, 5441-5444) that subunit VII is involved in electron transfer reactions at center o of the b-c1 complex. We suggest that the binding of antibody to subunit VII inhibits the transfer of electrons to cytochrome b 566. Consequently, two electrons are transferred to the iron-sulfur protein and cytochrome c1 through an antimycin-insensitive pathway. Moreover, the antibody may change the conformation of subunit VII, such that the myxothiazol and hydroxynaphthoquinone binding sites are partially blocked thus permitting electron flow to cytochrome c1. PMID- 2547778 TI - A human muscle adenine nucleotide translocator gene has four exons, is located on chromosome 4, and is differentially expressed. AB - The human heart-skeletal muscle adenine nucleotide translocator (ANT1) gene was isolated and sequenced. It spans 5.8 kilobases and contains four exons. The 5' nontranscribed region contains typical CCAAT and TATA sequences, a 22-nucleotide pair inverted repeat and a 13-nucleotide pair sequence homologous to a similar region in the ATP synthase beta subunit gene. The region surrounding the first exon and intron is G+C-region surrounding the first exon and intron is G+C-rich, and the intron contains three Sp1 binding motifs. ANT1 was assigned to chromosome 4 using both flow-sorted chromosomes and segregating human-mouse hybrid cells. Additional ANT sequences were found on at least two other chromosomes. ANT1 transcripts were present at high levels in human heart and skeletal muscle but were almost undetectable in liver, kidney, and brain. By contrast, fibroblast ANT (ANT2) mRNAs were present in all five tissues. The unique nature and arrangement of the ANT1 transcriptional control elements may account for this differential expression. PMID- 2547779 TI - A cAMP-responsive element regulates expression of the mouse steroid 11 beta hydroxylase gene. AB - In Y1 mouse adrenocortical tumor cells, expression of steroid 11 beta-hydroxylase (11 beta-OHase) is stimulated by cAMP following a delay of 4-6 h. Our results demonstrate that a cAMP-responsive element (CRE) within the 11 beta-OHase promoter region is a major determinant of this induction. The 5'-flanking sequences from the mouse 11 beta-OHase gene were placed in front of a human growth hormone reporter gene and transfected into Y1 cells. Treatment of transfected cells with 8-bromo-cAMP increased expression directed by the 11 beta OHase 5'-flanking region by 3.8-fold. In 5'-deletion analyses, 123 base pairs of 5'-flanking sequences were sufficient for cAMP induction, whereas cAMP treatment did not affect expression of a plasmid with only 40 base pairs of 5'-flanking sequence. Within these 123 base pairs, a region from -56 to -49 matched 7 of 8 bases comprising the consensus sequence for the CRE. 11 beta-OHase 5'-flanking sequences from -65 to -42, including the CRE-like sequence, conferred cAMP inducibility to promoters from the thymidine kinase and chorionic gonadotropin alpha-subunit genes. DNase I footprinting and Southwestern blotting analyses demonstrated that the protein which interacted with the CRE in the 11 beta-OHase promoter region was similar to the CRE-binding protein associated with other cAMP regulated genes. Together, these results suggest that an interaction between the 11 beta-OHase CRE and CRE-binding protein mediates cAMP induction of the 11 beta OHase gene. PMID- 2547781 TI - The inositol 1,4,5-trisphosphate-binding site in adrenal cortical cells is distinct from the endoplasmic reticulum. AB - The distribution of binding sites for the calcium-mobilizing second messenger inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) was investigated in subcellular fractions of bovine adrenal cortex. The [3H]Ins(1,4,5)P3-binding capacity was enriched in the microsomal fraction, which contained a single class of high affinity binding sites with a Kd of 21.6 +/- 3.0 nM. The specific [3H]Ins(1,4,5)P3 binding appeared to be sharply pH dependent and was inhibited by millimolar concentrations of ATP. Upon fractionation of microsomes on sucrose density gradient there was a clearcut separation of the Ins(1,4,5)P3 receptor containing fractions from those enriched in specific endoplasmic reticulum markers such as sulfatase C activity or RNA content. The microsomes enriched in Ins(1,4,5)P3-binding sites were of lower density than the endoplasmic reticulum and co-purified partly with the plasma membrane. In addition, Ins(1,4,5)P3 sensitive 45Ca2+ uptake into the microsomes was maximal in the lighter fractions. This distinction between Ins(1,4,5)P3-binding sites and endoplasmic reticulum derived microsomes was confirmed upon fractionation according to their electrophoretic mobilities by free flow electrophoresis. These results indicate that in adrenal cortical cells, the source of Ca2+ mobilized by Ins(1,4,5)P3 upon stimulation with an agonist is not located in the endoplasmic reticulum. Our data support the hypothesis that a specialized vesicular organelle, distinct from endoplasmic reticulum and in close apposition with the plasma membrane, is involved in intracellular Ca2+ homeostasis. PMID- 2547780 TI - Possible direct linkage of insulin-like growth factor-II receptor with guanine nucleotide-binding proteins. AB - In mouse Balb/c3T3 fibroblasts, insulin-like growth factor (IGF)-II activates a calcium-permeable cation channel through a cell surface IGF-II receptor (Kojima, I., Nishimoto, I., Iiri, T., Ogata, E., and Rosenfeld, R. G. (1988) Biochem. Biophys. Res. Commun. 154, 9-19; Matsunaga, H., Nishimoto, I., Kojima, I., Yamashita, N., Kurokawa, K., and Ogata, E. (1988) Am. J. Physiol. 255, C442 C446). In the action of IGF-II, a pertussis toxin (or islet-activating protein; IAP)-sensitive GTP-binding protein (G protein) is inferred to be involved (Nishimoto, I., Hata, Y., Ogata, E., and Kojima, I. (1987) J. Biol. Chem. 262, 12120-12126). In the present study, we examined the direct coupling of the IGF-II receptor with G proteins. In broken Balb/c3T3 cell membranes, 10 nM IGF-II rapidly attenuated the IAP-catalyzed ADP-ribosylation of a 40-kDa protein in a manner requiring magnesium ion. The IGF-II-mediated attenuation in the IAP substrate activity was 80% recovered after washing off IGF-II and inhibited by coexisting guanosine 5'-O-(2-thiodiphosphate), while either aluminum fluoride solution (10 mM NaF plus 100 microM AlCl3) or 100 microM guanosine 5'-O-(3 thiotriphosphate) (GTP gamma S) reproduced the action of IGF-II. When purified IAP substrate G proteins (Gi1, Gi2, G0) were incubated with IGF-II in the presence of membranes from IAP-treated Balb/c3T3 cells, the attenuation in the IAP substrate activity was evident in Gi2, but not in Gi1 or G0. On the other hand, 10 nM insulin had no effect on the modification of the 40-kDa IAP substrate in Balb/c3T3 cell membranes, whereas 10 nM IGF-I elicited a slow onset of the IAP sensitivity attenuation from the 40-kDa protein. However, the specific involvement of the IGF-II receptor in the modification of the IAP substrate induced by low concentrations of IGF-II was suggested by the observations that (i) IGF-I receptor-lacking cell membranes were effective for the Gi2 modification by IGF-II, (ii) the ability of membranes to mediate the action of IGF-II was markedly attenuated in IGF-II receptor-lacking cell membranes, and (iii) agonistic anti-IGF-II receptor antibody mimicked the action of IGF-II on the 40 kDa protein in Balb/c3T3 cell membranes in a dose-dependent manner similar to that observed in the antibody-induced blocking of membrane IGF-II binding.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2547782 TI - Cross-linking of the high affinity Fc receptor for human immunoglobulin G1 triggers transient activation of NADPH oxidase activity. Continuous oxidase activation requires continuous de novo receptor cross-linking. AB - Cross-linking of the high affinity Fc receptor for human immunoglobulin G1 (Fc gamma RI) on U937 cells triggered superoxide anion (O-2) release. This was accomplished by the binding of an Fc gamma RI-specific monoclonal antibody, mAb 32, followed by cross-linking of the mAb on the cell with anti-mouse IgG F(ab')2 by Fc gamma RI-specific mAbs 32 and 22 used as an equimolar mixture or by Fc gamma RI-specific mAb 197 (a murine IgG2a and thus a multivalent ligand for Fc gamma RI) alone. At subsaturating concentrations of the Fc gamma RI-cross-linking ligands, O2- generation was continuous over relatively long intervals. However, saturating concentrations triggered an often substantial but always transient O2- burst. This transient burst of oxidase activity ceased with maximal ligand accumulation on the cell. Cells in which oxidase activity had ceased could be restimulated using phorbol 12-myristate 13-acetate or aggregated human IgG1, indicating that cessation of O2- generation was not due to a generalized exhaustion or inhibition of the NADPH oxidase pathway. Cells incubated in subsaturating concentrations of cross-linking antibodies continued to release O2- until binding of the ligand ceased. In addition, the rates of O2- production and ligand accumulation were the same. Thus, continuous O2- production appeared to be dependent upon continuous de novo formation of cross-linked and activated Fc gamma RI. Furthermore, the mol of O2- released in response to Fc gamma RI cross linking by the multivalent ligand mAb 197 were directly proportional to the mol of mAb bound over a range of saturating and subsaturating concentrations. This evidence suggests a quantal relationship between each Fc gamma RI activated (cross-linked) and the resultant oxidase activity and supports a "rate" model for the activation of this response. Thus, each Fc gamma RI entering the pool of activated receptors probably makes a unitary contribution to the signal. An additional finding showed that cross-linked Fc gamma RI became associated with the cell cytoskeleton and that this association was also transient. Dissociation of Fc gamma RI from its cytoskeletal attachment occurred well after cessation of O2- production. PMID- 2547783 TI - Partial characterization of a glucocorticoid suppressible mitogenic activity secreted from a rat hepatoma cell line hypersensitive to the antiproliferative effects of glucocorticoids. AB - We have previously shown that glucocorticoids suppress the proliferation of Fu5 hepatoma cells and have selected subclones which are either hypersensitive (BDS1) or resistant (EDR3) to the antiproliferative effects of dexamethasone, a synthetic glucocorticoid. BDS1 cells externalize a glucocorticoid suppressible mitogenic activity (denoted GSM) which stimulated [3H]thymidine incorporation in quiescent, serum-starved Balb/c 3T3 cells. Glucocorticoid treatment of BDS1 cells reduced the secreted levels of GSM activity by approximately 20-fold in comparison to untreated cells. The GSM activity was constitutively secreted from a glucocorticoid receptor minus variant (EDR3) demonstrating that the suppression of this mitogenic activity is a new glucocorticoid hormone response which required a functional receptor. GSM activity was sensitive to sulfhydryl reducing agents or trypsin, stable to heat and acid treatments and fractionated in gel filtration columns with a native molecular weight of approximately Mr 30,000. The persistence of this size for mitogenic activity after electrophoretic fractionation in nonreducing sodium dodecyl sulfate-poly-acrylamide gels suggested that the GSM activity is comprised of a single protein. Total secreted protein isolated from untreated BDS1, but not dexamethasone-treated BDS1, stimulated 3T3 cells to grow in transformed-appearing large colonies in soft agar and to display multiple layering and elongated spindle-like morphology on solid substratum. The addition of both insulin and EGF to conditioned medium protein isolated from glucocorticoid-treated BDS1 cells restored full induction of 3T3 cell anchorage-independent growth while insulin restored full and EGF partial mitogenic stimulation of these fibroblasts. These results suggest that the GSM activity acts in a pathway common to that of insulin or EGF in fibroblasts. PMID- 2547784 TI - Granulocyte-macrophage colony-stimulating factor primes neutrophils by activating a pertussis toxin-sensitive G protein not associated with phosphatidylinositol turnover. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a hematopoietic cytokine which produces diverse biological effects in target cells of myeloid origin. GM-CSF enhances the production of superoxide anion (O2-) by mature neutrophils in response to chemotactic peptides such as formyl-methionyl-leucyl phenylalanine (fMLP), but alone it has no effect on this system. This process has been termed "priming." fMLP activates neutrophils via a pertussis toxin-sensitive GTP-binding protein, leading to the rapid production of the second messengers diacylglycerol (DAG) and inositol trisphosphate, via phosphatidylinositol turnover, and arachidonic acid (AA) by a presumptive phospholipase A2-mediated mechanism. All three second messengers may lead to the generation of O2-. We investigated the effect of priming of GM-CSF on these systems. GM-CSF had no effect on fMLP-stimulated DAG and inositol trisphosphate levels, nor did it amplify the response to exogenously added phorbol ester (to mimic the action of DAG) or calcium ionophore. Neutrophils primed with the cytokine showed a small, but significant, enhancement of fMLP-stimulated AA release. Compared with unprimed controls, primed neutrophils also showed a significant increase in O2- production when stimulated with either AA or the nonhydrolyzable GTP analogue, GTP-gamma-S. The magnitude of enhanced O2- production was similar to that observed after fMLP treatment of primed cells. All of these effects, including the increased sensitivity to AA treatment, were inhibited by pertussis toxin. These data show that GM-CSF primes neutrophils by modulating the activity of at least one pertussis toxin-sensitive G protein coupled to a metabolic pathway that mobilizes and utilizes arachidonic acid. PMID- 2547785 TI - Recombinant expression of a secreted form of the atrial natriuretic peptide clearance receptor. AB - A general structure for the atrial natriuretic peptide clearance receptor (ANP C receptor) has been proposed based on hydropathicity analysis of the deduced amino acid sequence of this membrane protein (Fuller, F., Porter, J.G., Arfsten, A., Miller, J., Schilling, J., Scarborough, R.M., Lewicki, J.A., and Schenk, D.B. (1988) J. Biol. Chem. 263, 9395-9401). The ANP C-receptor is believed to possess a large amino-terminal extracellular domain (436 amino acids), a single hydrophobic transmembrane anchor (23 amino acids), and a short cytoplasmic tail (37 amino acids). As a means of testing the structure and proposed cellular orientation of this protein, we have employed the technique of in vitro mutagenesis to prepare a receptor mutant (anc-) lacking the transmembrane and cytoplasmic domains. Expression of this mutant in mammalian cells using a vaccinia virus vector results in secretion of a truncated soluble form of the ANP C-receptor which binds native ANP and synthetic ANP analogs with a specificity similar to that of the native ANP C-receptor. In contrast to the native ANP C receptor that exists predominantly as a homodimer on the cell surface, the secreted receptor exists as a monomeric species. The results are consistent with the proposed structure of this receptor with the amino-terminal domain containing the ANP-binding site oriented extracellular to the plasma membrane. In addition, these data demonstrate that the receptor does not require association with the plasma membrane or its native dimeric configuration in order to bind ANP ligands with high affinity and specificity. PMID- 2547787 TI - Structure-function studies on bacteriorhodopsin. IX. Substitutions of tryptophan residues affect protein-retinal interactions in bacteriorhodopsin. AB - Bacteriorhodopsin contains 8 tryptophan residues distributed across the membrane embedded helices. To study their possible functions, we have replaced them one at a time by phenylalanine; in addition, Trp-137 and -138 have been replaced by cysteine. The mutants were prepared by cassette mutagenesis of the synthetic bacterio-opsin gene, expression and purification of the mutant apoproteins, renaturation, and chromophore regeneration. The replacement of Trp-10, Trp-12 (helix A), Trp-80 (helix C), and Trp-138 (helix E) by phenylalanine and of Trp 137 and Trp-138 by cysteine did not significantly alter the absorption spectra or affect their proton pumping. However, substitution of the remaining tryptophans by phenylalanine had the following effects. 1) Substitution of Trp-86 (helix C) and Trp-137 gave chromophores blue-shifted by 20 nm and resulted in reduced proton pumping to about 30%. 2) As also reported previously (Hackett, N. R., Stern, L. J., Chao, B. H., Kronis, K. A., and Khorana, H. G. (1987) J. Biol. Chem. 262, 9277-9284), substitution of Trp-182 and Trp-189 (helix F) caused large blue shifts (70 and 40 nm, respectively) in the chromophore and affected proton pumping. 3) The substitution of Trp-86 and Trp-182 by phenylalanine conferred acid instability on these mutants. The spectral shifts indicate that Trp-86, Trp 182, Trp-189, and possibly Trp-137 interact with retinal. It is proposed that these tryptophans, probably along with Tyr-57 (helix B) and Tyr-185 (helix F), form a retinal binding pocket. We discuss the role of tryptophan residues that are conserved in bacteriorhodopsin, halorhodopsin, and the related family of opsin proteins. PMID- 2547786 TI - Structure-function studies on bacteriorhodopsin. VIII. Substitutions of the membrane-embedded prolines 50, 91, and 186: the effects are determined by the substituting amino acids. AB - To study their role in the structure and function of bacteriorhodopsin, three prolines, presumed to be in the membrane-embedded alpha-helices, have been individually replaced as follows: Pro-50 and Pro-91 each by Gly and Ala and Pro 186 by Ala, Gly, and Val. The mutants of Pro-50 and Pro-91 all showed normal chromophore and proton pumping. However, the rates of regeneration of the chromophore in Pro-50----Ala, Pro-91----Ala and ----Gly with all-trans-retinal were about 30-fold slower than that in the wild-type, whereas the chromophore regeneration rate in Pro-50----Gly was 10-fold faster than in the wild-type. While, Pro-186----Ala regenerated the wild-type chromophore, the mutants Pro-186- --Val and Pro-186----Gly showed large blue shifts (about 80 nm) in the chromophore regenerated with all-trans-retinal and showed no apparent dark-light adaptation. Pro-186----Gly first regenerated the wild-type chromophore with 13 cis-retinal which was thermally unstable and rapidly converted to the blue shifted chromophore obtained with all-trans-retinal. High salt concentration restored the wild-type purple chromophore in the Pro-186----Gly mutant. Thus, in this mutant, the protein interconverts between two conformational states. Pro-186 ---Ala and Pro-186----Gly showed about 65%, whereas Pro-186----Val showed 10-20% of the normal proton pumping. PMID- 2547788 TI - Structure-function studies on bacteriorhodopsin. X. Individual substitutions of arginine residues by glutamine affect chromophore formation, photocycle, and proton translocation. AB - We have individually replaced all 7 of the arginine residues in bacteriorhodopsin by glutamine. The mutants with substitutions at positions 7, 164, 175, and 225 showed essentially the wild-type phenotype in regard to chromophore regeneration, chromophore lambda max, and proton pumping, although the mutant Arg-175----Gln showed decreased rate of chromophore regeneration. Glutamine substitutions of Arg 82, -134, and -227 affected proton pumping ability, and caused specific alterations in the bacteriorhodopsin photocycle. Finally, electrostatic interactions are proposed between Arg-82 and -227, and specific carboxylic acid residues in helices C and G, which regulate the purple to blue transition and proton transfers during the photocycle. PMID- 2547789 TI - Resonance Raman spectra of bovine liver catalase compound II. Similarity of the heme environment to horseradish peroxidase compound II. AB - Resonance Raman spectroscopy has been used to investigate the structure and environment of the heme group in bovine liver catalase compound II. Both Soret- and Q-band excitation have been employed to observe and assign the skeletal stretching frequencies of the porphyrin ring. The oxidation state marker band v4 increases in frequency from 1373 cm-1 in ferricatalase to 1375 cm-1 in compound II, consistent with oxidation of the iron atom to the Fe(IV) state. Oxidation of five-coordinate, high-spin ferricatalase to compound II is accompanied by a marked increase of the porphyrin core marker frequencies that is consistent with a six-coordinate low-spin state with a contracted core. An Fe(IV) = O stretching band is observed at 775 cm-1 for compound II at neutral pH, indicating that there is an oxo ligand at the sixth site. At alkaline pH, the Fe(IV) = O stretching band shifts to 786 cm-1 in response to a heme-linked ionization that is attributed to the distal His-74 residue. Experiments carried out in H218O show that the oxo ligand of compound II exchanges with bulk water at neutral pH, but not at alkaline pH. This is essentially the same behavior exhibited by horseradish peroxidase compound II and the exchange reaction at neutral pH for both enzymes is attributed to acid/base catalysis by a distal His residue that is believed to be hydrogen-bonded to the oxo ligand. Thus, the structure and environment of the heme group of the compound II species of catalase and horseradish peroxidase are very similar. This indicates that the marked differences in their reactivities as oxidants are probably due to the manner in which the protein controls access of substrates to the heme group. PMID- 2547790 TI - Phosphorylation of the alpha subunit of rat brain sodium channels by cAMP dependent protein kinase at a new site containing Ser686 and Ser687. AB - The alpha subunit of the rat brain sodium channel is phosphorylated by cAMP dependent protein kinase in vitro and in situ at multiple sites which yield seven tryptic phosphopeptides. Phosphopeptides 1-4 and 7 are derived from phosphorylation sites between residues 554 and 623 in a single large CNBr fragment from the cytoplasmic segment connecting homologous domains I and II of the alpha subunit (Rossie, S., Gordon, D., and Catterall, W. A. (1987) J. Biol. Chem. 262, 17530-17535). In the present work, antibodies were prepared against a synthetic peptide corresponding to residues 676-692 (AbSP15), which contain one additional potential phosphorylation site at Ser686-Ser687 in a different predicted CNBr fragment of this same intracellular segment. AbSP15 recognizes native and denatured sodium channels specifically and immunoprecipitates phosphorylated CNBr fragments of low molecular mass that contain a new site phosphorylated by cAMP-dependent protein kinase. Comparison of tryptic phosphopeptides derived from intact alpha subunits with those derived from the phosphorylated CNBr fragments isolated by immunoprecipitation with AbSP15 indicates that the two previously unidentified phosphopeptides 5 and 6 derived from the intact alpha subunit arise from phosphorylation of the site containing Ser686-Ser687. These results identify a new cAMP-dependent phosphorylation site and show that the major cAMP-dependent phosphorylation sites of the rat brain sodium channel, which are phosphorylated both in vitro and in intact neurons, are all located in a cluster between residues 554 and 687 in the intracellular segment between domains I and II. PMID- 2547791 TI - Relative efficiency of utilization of promoter and termination sites by bacteriophage T3 RNA polymerase. AB - Bacteriophage T3 RNA polymerase promoters have been classified as class II and class III on the basis of their relative location in T3 DNA as well as on the function of the protein products encoded by the messages transcribed from them. In the present work, the efficiency of utilization of several class II and class III promoters by bacteriophage T3 RNA polymerase was compared with regard to (a) rate of initiation of transcription as determined by [32P]PPi exchange with GTP; (b) complex formation between polymerase and promoters in the presence of GTP; and (c) competition between different promoters for T3 RNA polymerase in a standard transcription assay. The results of these experiments indicated that the class II promoters at 1.05 and 22.8 T3 map units, whose promoter sequences are remarkably similar to the consensus class III promoter sequences, are nearly as strong as typical class III promoters. In contrast, the class II promoter at 14.3 T3 map units, whose promoter sequence differs from the consensus class III promoter sequence by having a C:G base pair instead of a usual A:T base pair at the -1 position, was considerably weaker than the class III promoter. When the C:G base pair at this position was changed to A:T using site-directed mutagenesis, the rate of initiation of RNA synthesis from the mutant promoter was similar to that of a typical class III promoter. In agreement with this observation, it was observed that changing the A:T base pair at the -1 position of a strong class II promoter, at 1.05 T3 map units, to C:G decreased the rate of RNA synthesis from this promoter by about 65%. These observations indicate that the nucleotide residues at the -1 position play a critical role in determining the efficiency of promoter utilization by T3 RNA polymerase. The two termination sites recognized in vitro by bacteriophage T3 RNA polymerase on the T3 genome have been cloned, sequenced, and mapped. Analysis of the DNA nucleotide sequence surrounding the termination site at 59.7 map units indicated that the putative RNA transcript arising from this region can be arranged into a GC-rich stem-loop structure followed by a U-rich 3' tail. However, a major fraction of T3 RNA polymerase molecules read through this terminator in vitro to transcribe regions of T3 DNA beyond this terminator. In contrast to termination at 59.7 map units, termination of transcription at 100 T3 map units does not occur in response to any putative terminator structure or sequence.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2547792 TI - The receptor for alpha-melanotropin of mouse and human melanoma cells. Application of a potent alpha-melanotropin photoaffinity label. AB - The melanotropin (MSH) receptor of mouse B16-F1 melanoma cells was characterized by photoaffinity cross-linking, using a potent alpha-MSH photolabel, [norleucine4, D-phenylalanine7, 1'-(2-nitro-4-azidophenylsulfenyl)-tryptophan9] alpha-melanotropin (Naps-MSH). Its monoiodinated form, 125I-Naps-MSH, displayed a approximately 6.5-fold higher biological activity than alpha-MSH. Scatchard analysis of the saturation curves with 125I-Naps-MSH revealed approximately 20,000 receptors/B16-F1 cell and an apparent KD of approximately 0.3 nM. Analysis of the cross-linked MSH receptor by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that a photolabeled band of approximately 45 kDa occurs in B16-F1, B16-F10, and Cloudman S91 mouse melanoma, as well as in human D10 and 205 melanoma but not in non-melanoma cells. The labeled 45-kDa protein had an isoelectric point of 4.5-4.9 as determined by two-dimensional gel electrophoresis. Treatment of the labeled 45-kDa protein of B16-F1 cell membranes by neuraminidase shifted the band to approximately 42 kDa. A similar band of about 42 kDa was also observed after receptor labeling of B16-W4 cells, a cell line with a decreased number of terminal N-linked neuraminyl residues. These results indicate that the labeled 45-kDa glycoprotein contains terminal sialic acid residues, explaining the low pI of this protein, and that it is characteristic for melanoma cells and hence part of the MSH receptor. PMID- 2547793 TI - In situ phosphorylation of platelet actin-binding protein by cAMP-dependent protein kinase stabilizes it against proteolysis by calpain. AB - To identify the protein kinase that is responsible for catalyzing phosphorylation of actin-binding protein (ABP) in platelets, we have examined the effects of protein kinase C and cAMP-dependent protein kinase on this process. We found that purified platelet protein kinase C from platelets was unable to phosphorylate ABP in vitro. However, a crude platelet kinase preparation phosphorylated ABP in the presence of cAMP, but not in the presence of Ca2+/phosphatidylserine. Fresh platelet plasma membranes incubated with [gamma-32P]ATP phosphorylated ABP in the presence of cAMP and the process was blocked by a cAMP-dependent protein kinase inhibitor; ABP phosphorylation induced by prostaglandin E1 (PGE1) appeared to be reduced by the subsequent addition of thrombin. These results strongly suggest that in situ ABP is phosphorylated by activated cAMP-dependent protein kinase when platelet function is inhibited by PGE1. Furthermore, in the PGE1-treated platelets, ABP was proteolyzed at a slower rate than in control platelets when they were lysed with Triton in the absence of EGTA. Partially purified ABP was proteolyzed by calpain in vitro at a slower rate as well. It was demonstrated that ABP from PGE1-treated platelets recovered its sensitivity to calpain after ABP was incubated with a protein phosphatase that had been purified from platelets. We postulate that ABP is stabilized against proteolysis in response to cAMP-elevating agents and that this blocks cytoskeleton reorganization. PMID- 2547794 TI - Control of cytosolic free calcium in rat and chicken osteoclasts. The role of extracellular calcium and calcitonin. AB - Single cell [Ca2+], studies were performed in chicken and rat osteoclasts loaded with fura-2 and exposed to a variety of treatments. Under resting conditions, basal [Ca2+]i, was 79.2 +/- 47.3 and 84.3 +/- 65.7 nM (averages +/- S.D.; n = 141 and 126) in the osteoclasts of the two species, respectively. Basal [Ca2+]i was stable in all rat and in approximately 80% of chicken osteoclasts. In the remaining 20%, spontaneous, irregular [Ca2+], fluctuations were observed (amplitude range: 50-200 nm over basal values). Increase of [Ca2+]o over the concentration of the Krebs-Ringer incubation medium (2 mM) induced rises of [Ca2+] in almost all cells investigated. [Ca2+] rises were already appreciable with 0.5 mM [Ca2+]o additions and reached high values with 4 mM additions: 390 +/ 113 and 364 +/- 214 nM [Ca2+], in rat and chicken osteoclasts, respectively (n = 122 and 101). Qualitatively, the responses to [Ca2+]o additions consisted of discrete [Ca2+]i transients, biphasic (an initial spike followed by a plateau), or monophasic (either the spike or the plateau). In a few chicken osteoclasts, the [Ca2+]i increase occurring after [Ca2+]o addition consisted of multiple, irregular fluctuations, similar to those observed in 20% of these cells under resting conditions. In individual osteoclasts subsequently exposed to multiple [Ca2+]o increase pulses, the type of the [Ca2+]i transient (mono- or biphasic) was maintained, and the size was dependent on the magnitude of the [Ca2+]o additions. Effects similar to those of [Ca2+]o were induced by the addition of Cd2+ or Ba2+ (but not La3+ or Mg2+) into the medium. The Cd2+ effect was maintained in part even in a Ca2+-free medium. Of various hormones and factors, parathormone, 1,25-dihydroxyvitamin D3, and prostaglandin E2 were inactive. In contrast, calcitonin was active in rat osteoclasts (which express numerous receptors). [Ca2+]i increases were small (19 +/- 17.9 nM; n = 21) when the hormone was administered alone; they were synergistic (severalfold potentiation) when the hormone was administered before or after [Ca2+]o. The [Ca2+]i effects of calcitonin were mimicked by 8Br-cAMP (31 +/- 26 nM; n = 12) when the nucleotide was administered alone; marked synergism when it was administered in combination with [Ca2+]o. This paper demonstrates for the first time that changes of [Ca2+]i are induced in osteoclasts by treatments with [Ca2+]o and calcitonin and can therefore be involved in intracellular mediation of the physiological effects of these two extracellular signals. PMID- 2547795 TI - Synaptic vesicle ceramide kinase. A calcium-stimulated lipid kinase that co purifies with brain synaptic vesicles. AB - Much current work on the mechanism of neurosecretion has focused on proteins specific to neural secretory vesicles (synaptic vesicles). We report a calcium stimulated lipid kinase that co-purifies with rat brain synaptic vesicles. This enzyme activity is found only in membrane fractions that contain synaptic vesicle markers. Based on identification of the lipid product as ceramide 1-phosphate and on the finding that ceramide kinase activity co-purifies with synaptic vesicles, the enzyme is proposed to be a ceramide kinase. Kinase activity is stimulated by micromolar concentrations of calcium. Calcium increases the apparent Vmax of the reaction with little effect on the Km for ceramide. The vesicular localization of this enzyme, the requirement for ATP, and the stimulation of enzyme activity by micromolar calcium suggest that ceramide phosphorylation may be associated with neurotransmitter release. PMID- 2547796 TI - Interaction of nuclear factors with multiple sites in the somatic cytochrome c promoter. Characterization of upstream NRF-1, ATF, and intron Sp1 recognition sequences. AB - The rat somatic cytochrome c promoter is resolved into a mosaic of cis-acting upstream and intron elements required for maximal activity. Mutations in each diminished cytochrome c promoter activity and eliminated the specific binding of cognate nuclear factors. Among these is the recognition sequence for a nuclear factor designated NRF-1 (nuclear respiratory factor 1) also found in the upstream regions of several other nuclear genes whose products function in the mitochondria. The NRF-1 site was tightly coupled to a second functionally independent element (region I), and together these sites constitute a major determinant of cytochrome c expression. In addition to these novel sequence elements, the promoter also contained recognition sites for the common transcriptional activators ATF and Sp1. A potent promoter element within the first intron consisted of two adjacent Sp1 binding sites. Point mutations in the first site eliminated the promoter activity of the element as well as Sp1 binding to both sites. An ATF recognition sequence in the upstream promoter was identical to an authentic cyclic AMP (cAMP) responsive element in stimulating promoter activity and in conferring a cAMP response upon a heterologous promoter. These promoter elements and their cognate nuclear factors likely contribute to the housekeeping function of cytochrome c and to the coordinate modulation of respiratory gene expression according to cellular energy demands. PMID- 2547798 TI - Cyclic AMP-dependent protein kinase in Paramecium tetraurelia. Its purification and the production of monoclonal antibodies against both subunits. AB - Two soluble cAMP-dependent protein kinases were purified from the cytoplasm of Paramecium tetraurelia. Both kinases consisted of a 40-kDa catalytic subunit and a 44-kDa regulatory subunit. The two forms of the enzyme were separated by anion exchange chromatography. Affinity chromatography on cAMP-Sepharose separated the regulatory subunit (retained by the column) from the cAMP-independent catalytic subunit (not retained). Four classes of monoclonal antibodies were generated. One class was specific for the catalytic subunit of both cAMP-dependent protein kinases, and three classes recognized the regulatory subunit of both forms of the enzyme. Subunits of 40 and 44 kDa were detected on immunoblots of purified cilia and of crude cell extracts. In addition, one class of antibodies specific for the regulatory subunit detected a ciliary protein with a molecular mass of 48 kDa. The monoclonal antibodies did not recognize type I or type II cAMP-dependent protein kinase from rabbit muscle nor did they cross-react with proteins from several unicellular eucaryotes, with one exception: antibodies specific for the catalytic subunit recognized a 40-kDa protein of Tetrahymena pyriformis. PMID- 2547797 TI - Modulation of T cell activation by differential regulation of the phosphorylation of two cytosolic proteins. Implication of both Ca2+ and cyclic AMP-dependent protein kinases. AB - Interleukin 2 production by activated Jurkat T cells is markedly decreased by prostaglandin E2 (PGE2). The target of PGE2 action has been investigated in the present study. Among the biochemical events occurring after CD3.TCR triggering by anti-CD3 monoclonal antibody, phosphorylation of two cytosolic proteins, pp21 and pp23, was strongly inhibited by PGE2, forskolin, and 8-bromo-cAMP, whereas anti CD3 monoclonal antibody-induced CD3.TCR modulation and Ca2+ influx were not affected. The inhibition of both pp21 and pp23 phosphorylation and interleukin 2 synthesis by PGE2 can be largely reversed by the cAMP-dependent protein kinase inhibitor, N-[2-(methylamino)-ethyl-1]-5-isoquinoline sulfonamide. Together with the demonstration of a cAMP-dependent protein kinase activity in Jurkat T cells, these results are consistent with the participation of the cAMP-dependent protein kinase mediating the inhibitory action of PGE2, probably through the inhibition of pp21 and pp23 phosphorylation. Thus, it appears that the modulation of the phosphorylation of these cytosolic proteins represents an essential step in the regulation of T lymphocyte activation. PMID- 2547799 TI - The Escherichia coli primosome can translocate actively in either direction along a DNA strand. AB - The primosome is a mobile multiprotein DNA replication-priming apparatus that requires seven Escherichia coli proteins (replication factor Y (protein n'), proteins n and n", and the products of the dnaB, dnaC, dnaT, and dnaG genes) for assembly at a specific site (termed a primosome assembly site) on single-stranded DNA binding protein-coated single-stranded DNA. Two of the protein components of the primosome have intrinsic DNA helicase activity. The DNA B protein acts in the 5'----3' direction, whereas factor Y acts in the 3'----5' direction. The primosome complex has DNA helicase activity when present at a replication fork in conjunction with the DNA polymerase III holoenzyme. In this report, evidence is presented that the multiprotein primosome per se can act as a DNA helicase in the absence of the DNA polymerase III holoenzyme. The primosome DNA helicase activity can be manifested in either direction along the DNA strand. The directionality of the primosome DNA helicase activity is modulated by the concentration and type of nucleoside triphosphate present in the reaction mixture. This DNA helicase activity requires all the preprimosomal proteins (the primosomal proteins minus the dnaG-encoded primase). Preprimosome complexes must assemble at a primosome assembly site in order to be loaded onto the single-stranded DNA and act subsequently as a DNA helicase. The 5'----3' primosome DNA helicase activity requires a 3' single-stranded tail on the fragment to be displaced, while the 3'- --5' activity does not require a 5' single-stranded tail on the fragment to be displaced. Multienzyme preprimosomes moving in either direction are capable of associating with the primase to form complete primosomes that can synthesize RNA primers. PMID- 2547800 TI - Molecular basis for resistance to myxothiazol, mucidin (strobilurin A), and stigmatellin. Cytochrome b inhibitors acting at the center o of the mitochondrial ubiquinol-cytochrome c reductase in Saccharomyces cerevisiae. AB - The respiratory bc1 complex transfers the electrons from ubiquinol to cytochrome c oxidase. Myxothiazol, strobilurin A (mucidin), and stigmatellin are center o inhibitors preventing electron transfer at the ubiquinone redox site Qo, which is located closer to the outer side of the inner mitochondrial membrane. The cytochrome b gene is carried by the organelle DNA. Yeast mutants resistant to myxothiazol and mucidin have been previously isolated and mapped to specific loci of the cytochrome b gene. In the present work, stigmatellin-resistant mutants were isolated and genetically analyzed. The mutated amino acid residues from seven myxothiazol-, four mucidin-, and six stigmatellin-resistant mutants have been identified by sequencing the relevant segments of the resistant cytochrome b gene. A third myxothiazol-resistant locus and the first stigmatellin-resistant locus were identified. The mutated codons were found to be clustered in two regions of the cytochrome b protein which appeared to be responsible for the resistance to Qo site inhibitors. The first region is within the end of the first, the second, and the beginning of the third exon whereas the second region is within exon five and the beginning of the sixth exon. PMID- 2547801 TI - Evidence for ecto-protein kinase activity that phosphorylates Kemptide in a cyclic AMP-dependent mode. AB - The heptapeptide Leu-Arg-Arg-Ala-Ser-Leu-Gly (Kemptide) is a synthetic construct of a substrate for cAMP-dependent protein kinase (PK). In this work we show that Kemptide has all the properties of a cytophilic substrate, i.e. it is a molecule preserving cell membrane intactness when added to cultured cells. Kemptide thus satisfies the prerequisites for employment in assays for cell surface-located ecto-PK activity. Different types of intact cells catalyze the phosphorylation of Kemptide in the presence of extracellular ATP and cAMP with Km values of 3-4 microM for Kemptide. Kemptide phosphorylation was influenced by PKI, the inhibitory protein specific for cAMP-PK. The results of comparative experiments with intact cells and with cell extracts demonstrate the ectoenzyme nature of this cAMP-PK. Further, the possibility was ruled out of a transfer of enzyme activity from damaged cells to the surface of intact cells. The anchorage of the surface cAMP-PK activity to the plasma membrane appears to be relatively stable since (i) cell supernatants, obtained after preincubation of intact cells with cAMP or Kemptide, did not show Kemptide phosphorylation, and (ii) the cAMP dependent PK activity remained with cells even after five consecutive washes with cAMP or Kemptide. This is in contrast to the ecto-cAMP-independent phosvitin/casein type PK (Kubler, D., Pyerin, W., Burow, E., and Kinzel, V. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 4021-4025) which is released from intact cells through the addition of substrate. Data are presented which show that both ectokinase activities are exhibited independently. In conjunction with published evidence for an active export of cAMP from cells as well as for the appearance of extracellular ATP the demonstration of an ecto-cAMP-PK further supports the potential of PK for intercellular regulation. The potential of ecto-cAMP-PK is demonstrated by its ability to phosphorylate biologically active forms of atrial natriuretic peptide, the atrial natriuretic peptide, which possesses the specific sequence for a cAMP-PK-catalyzed phosphorylation. PMID- 2547802 TI - Neuron-specific enolase can be used as the sole guide to treat small-cell lung cancer patients in common clinical practice. AB - Serum samples were collected from 115 small-cell lung cancer patients before each course of chemotherapy and during follow-up. Levels of neuron-specific enolase (NSE) were measured and compared to the clinical assessments of the course of the disease, which were done by the responsible physician without knowledge of NSE values. The predictive accuracy of an increase or decrease of NSE for a major response (CR + PR), SD or PD was 98%. Importantly no false-positive rises of NSE were observed. On the basis of this large number of data it seems justified to conclude that in common clinical practice the treatment of small-cell lung cancer patients can be monitored by serial measurements of NSE alone. PMID- 2547803 TI - Transformation by Rous sarcoma virus induces clathrin heavy chain phosphorylation. AB - We have shown that the heavy chain of clathrin is phosphorylated in chicken embryo fibroblast cells transformed by Rous sarcoma virus, but not in normal cells. Approximately 1 mol of phosphate is bound for every 5 mol of heavy chain in the maximally phosphorylated transformed cells. Two-thirds of the phosphate is on serine and one-third on tyrosine residues. Clathrin heavy chain is a substrate for pp60v-src in vitro. Cleveland analysis of the in vivo and in vitro clathrin heavy chain phosphopeptides, generated by protease V8 digestion, show labeled proteolytic fragments of similar molecular weight, suggesting that pp60v-src could be directly responsible for the in vivo phosphorylation of clathrin. Phosphate is equally incorporated into clathrin in both the unassembled and the assembled clathrin pools, whereas [35S]methionine is preferentially incorporated into the assembled pool. In normal cells, clathrin visualized by immunofluorescent staining appears in a punctate pattern along the membrane surface and concentrated around the nucleus; in transformed cells the perinuclear staining is completely absent. The phosphorylation of clathrin heavy chain in transformed cells may be linked to previously observed transformation-dependent alterations in receptor-mediated endocytosis of ligands such as EGF and thrombin. PMID- 2547804 TI - Expression of human plasma gelsolin in Escherichia coli and dissection of actin binding sites by segmental deletion mutagenesis. AB - Human plasma gelsolin has been expressed in high yield and soluble form in Escherichia coli. The protein has nucleating and severing activities identical to those of plasma gelsolin and is fully calcium sensitive in its interactions with monomeric actin. A number of deletion mutants have been expressed to explore the function of the three actin binding sites. Their design is based on the sixfold segmental repeat in the protein sequence. (These sites are located in segment 1, segments 2-3, and segments 4-6). Two mutants, S1-3 and S4-6, are equivalent to the NH2- and COOH-terminal halves of the molecule obtained by limited proteolysis. S1-3 binds two actin monomers in the presence or absence of calcium, it severs and caps filaments but does not nucleate polymerization. S4-6 binds a single actin monomer but only in calcium. These observations confirm and extend current knowledge on the properties of the two halves of gelsolin. Two novel constructs have also been studied that provide a different pairwise juxtaposition of the three sites. S2-6, which lacks the high affinity site of segment 1 (equivalent to the 14,000-Mr proteolytic fragment) and S1,4-6, which lacks segments 2-3 (the actin filament binding domain previously identified using the 28,000-Mr proteolytic fragment). S2-6 binds two actin monomers in calcium and nucleates polymerization; it associates laterally with filaments in the presence or absence of calcium and has a weak calcium-dependent fragmenting activity. S1,4 6 also binds two actin monomers in calcium and one in EGTA, has weak severing activity but does not nucleate polymerization. A model is presented for the involvement of the three binding sites in the various activities of gelsolin. PMID- 2547806 TI - Separation and indirect detection of alkyl sulfonates and sulfates. AB - Iron(II) 1,10-phenanthroline, Fe(phen)3(2+), salts are used as mobile phase additives for the liquid chromatographic separation of alkyl sulfonates and sulfates on the reversed-phase PRP-1. As alkyl chain length increases retention increases. For a given chain length an alkyl sulfate is more retained than the corresponding alkyl sulfonate. Major elution variables that affect retention are mobile phase solvent and counteranion concentration. Indirect photometric detection is used to detect alkyl sulfonates and sulfates at 510 nm where Fe(phen)3(2+) salts absorb. Conditions for isocratic and gradient elution of multicomponent mixtures are described. Detection limits depending on analyte approached 0.1 nmol for isocratic elution and 3 nmol for gradient elution. PMID- 2547807 TI - Effects of lipofuscin on in situ hybridization in human neuronal tissue. AB - In situ hybridization is a highly sensitive technique for detecting nucleic acid sequences within tissues, and is frequently employed in neurovirology. However, this technique requires many appropriate controls in order to recognize and avoid potential artifactual hybridization. We have encountered abundant reaction to lipofuscin in neurons in human peripheral and central nervous systems, using various DNA probes, which could be misinterpreted as positive signals. This pseudohybridization reaction was resistant to treatment with RNase or DNase and was also present in tissue sections treated with hybridization mixture or nuclear autoradiographic emulsion in the absence of any radioactive probes. Characteristics used to distinguish between authentic in situ hybridization and the reaction to neuronal lipofuscin include cellular localization, color, margins and granular appearance, sensitivity to treatment with nucleases and the effect of exposure time on signal intensity. These guidelines should be used to avoid potential misinterpretation of in situ hybridization results with human tissue. PMID- 2547805 TI - Signal transduction through the fibronectin receptor induces collagenase and stromelysin gene expression. AB - We have investigated the effects of ligation of the fibronectin receptor (FnR) on gene expression in rabbit synovial fibroblasts. Monoclonal antibodies to the FnR that block initial adhesion of fibroblasts to fibronectin induced the expression of genes encoding the secreted extracellular matrix-degrading metalloproteinases collagenase and stromelysin. That induction was a direct consequence of interaction with the FnR was shown by the accumulation of mRNA for stromelysin and collagenase. Monoclonal antibodies to several other membrane glycoprotein receptors had no effect on metalloproteinase gene expression. Less than 2 h of treatment of the fibroblasts with anti-FnR in solution was sufficient to trigger the change in gene expression, and induction was blocked by dexamethasone. Unlike other inducers of metalloproteinase expression, including phorbol diesters and growth factors, addition of the anti-FnR in solution to cells adherent to serum derived adhesion proteins or collagen produced no detectable change in cell shape or actin microfilament organization. Inductive effects were potentiated by cross linking of the ligand. Fab fragments of anti-FnR were ineffective unless cross linked or immobilized on the substrate. Adhesion of fibroblasts to native fibronectin did not induce metallo-proteinases. However, adhesion to covalently immobilized peptides containing the arg-gly-asp sequence that were derived from fibronectin, varying in size from hexapeptides up to 120 kD, induced collagenase and stromelysin gene expression. This suggests that degradation products of fibronectin are the natural inductive ligands for the FnR. These data demonstrate that signals leading to changes in gene expression are transduced by the FnR, a member of the integrin family of extracellular matrix receptors. The signaling of changes in gene expression by the FnR is distinct from signaling involving cell shape and actin cytoarchitecture. At least two distinct signals are generated: the binding of fibronectin-derived fragments and adhesion-blocking antibodies to the FnR triggers events different from those triggered by binding of the native fibronectin ligand. Because the genes regulated by this integrin are for enzymes that degrade the extracellular matrix, these results suggest that information transduced by the binding of various ligands to integrins may orchestrate the expression of genes regulating cell behavior in the extracellular environment. PMID- 2547808 TI - Determination of rotavirus serotype-specific antibodies in sera by competitive enhanced enzyme immunoassay. AB - A method is described for the specific detection of antibody to individual rotavirus serotypes in sera. A competitive enzyme immunoassay (EIA) was developed in which rotavirus serotype-specific monoclonal antibodies against VP7 compete with antibodies in test sera for rotavirus serotype-specific antigen bound to a solid phase. There was an excellent correlation between serotype-specific EIA results and serotype-specific neutralization titres (r = 0.915, P = less than 0.001). The value of this method for rotavirus epidemiology and vaccine trials is discussed. PMID- 2547809 TI - Factors affecting virus plaque confirmation procedures. AB - Direct plaque counts obtained by using the monolayer cell culture assay technique reliably confirmed the number of viruses isolated. Analysis revealed some significant differences in the false-positive rate, depending on the test method used or virus samples evaluated. Plaques from laboratory stock viruses showed a higher confirmation rate than sewage plaque isolates. Test results with laboratory stock viruses suggested that confirmation rates may be affected by virus types present in the sample. Plaques picked by the stab and scrape method and immediately passed into cell culture tubes produced the most reliable counts as compared to those picked by the stab only method or those stored at -70 degrees C in Earle's balanced salt solution with or without fetal calf serum. Plaque confirmation using this method was 90% or better. Although the term 'false positive plaque' has been applied to a particular plaque that was not confirmed, five of ten plaques picked by the stab and scrape method in one series of experiments were confirmed when repicked by the same method from original plaque bottles, indicating that a substantial number of unconfirmed plaques may be caused by plaque transfer techniques. PMID- 2547810 TI - Filter in situ hybridisation: an evaluation of the FISH technique for HPV detection in cervical swabs. AB - The filter in situ hybridisation (FISH) method for detection of HPV in cervical swabs was evaluated against the Southern blot technique on concomitant cervical biopsies. Of 73 biopsies, HPV 16 DNA sequences were found in 26 biopsies and HPV 18 sequences in 2 biopsies. Analysis by FISH of the corresponding smears detected 58 and 100% of these, respectively. Of the smears corresponding to the HPV negative biopsies, 17% were HPV 16-positive and 3% were HPV-18 positive by FISH. Re-hybridisation with cold plasmid added for competition did not change these results. To estimate the risk of spurious hybridisation between vector remnants in the probe and bacterial DNA sequences present in smears, we have hybridised by FISH to preparations of the 19 most common vaginal microorganisms. Of these, E. coli, which is present in about 10% of cervical smears, hybridised strongly with a probe of the plasmid vector pBR322 and may be a significant cause of false positive FISH results. None of the bacteria hybridised with probes of purified HPV when cold, denatured plasmid was added for competition. Analysis by FISH with probes of purified pBR322 to 167 smears of a patient control group resulted in 6% positive reactions. In hybridisations with probes of HPV 16 and 18 to 2 or 3 different filter preparations of the same smear, identical results were obtained in 18 of 19 smears, indicating a good reproducibility by the FISH method. The high percentage of HPV negative smears is equivalent to the rates known from cytology and may reflect sampling errors. PMID- 2547811 TI - An improved method for the concentration of rotaviruses from large volumes of water. AB - A microporous filter method for the concentration and reconcentration of rotaviruses from large volumes of water was developed. This method allows for the processing of large volumes of water with high recoveries of rotaviruses. Averages of 49% and 41% of seeded viruses were recovered from 20 and 400 l of water, respectively. Of the various eluents evaluated, a mixture of 10% tryptose phosphate broth and 3% beef extract at a pH value of 10.0 was found to give the greatest elution efficiency. The use of the positively charged 1MDS filters allowed for the processing of waters without the need for pH adjustments. An overall recovery of 35% was obtained from 400 l when hydroextraction was used as a second-step concentration procedure. PMID- 2547812 TI - Human cervical carcinoma cell lines contain an antigen identical to the tumor specific 75 kDa antigen of HeLa cells: detection by viral acquisition. AB - Purified vesicular stomatitis virus grown in the human cervical carcinoma HeLa cell line, VSV(HeLa), contains a 75 kDa tumor-specific antigen, detectable by immunoblotting of electrophoretically separated proteins with rabbit antiserum made against whole HeLa cells. Nearly identical results were obtained with VSV grown in the tumorigenic human hybrid ESH-5L cells, but not with the matched non tumorigenic ESH-5E cells. Growth of VSV in 4 other independently isolated human cervical carcinoma cell lines led to the concentration of the same 75 kDa tumor specific antigen by VSV. Infection of 2 other human cervical carcinoma cell lines did not lead to the detection of this antigen. The expression of the tumor specific antigen correlated directly with the amount of RNA expression from human papillomavirus integrated in the DNA of these cells, irrespective of whether the papillomavirus was type 16 or 18. PMID- 2547813 TI - Monoclonal blocking ELISA detecting serum antibodies to the glycoprotein gII of Aujeszky's disease virus. AB - A monoclonal blocking enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to Aujeszky's disease virus (ADV) in porcine serum was developed. This ELISA is based on the reaction between virus antigen immobilized in a microdilution plate and a monoclonal antibody (MAb) reactive with a highly stable epitope on a glycoprotein complex, gII, of ADV. The viral epitope was expressed by 18 European field, laboratory and vaccine strains of ADV. The MAb used in the test was selected among 15 MAbs all reactive with viral epitopes apparently recognized by the porcine immune system as well. Good agreement was found when serum samples from 375 pigs were tested in both a polyclonal and the monoclonal blocking ELISA. PMID- 2547814 TI - In situ hybridization: an optimised detection protocol for a biotinylated DNA probe renders it more sensitive than a comparable 35S-labelled probe. AB - The development of a highly sensitive biotinylated in situ hybridization protocol using egg drop syndrome virus infection of domestic fowl as a model is described. This protocol incorporates the use of a monoclonal antibody to biotin as the initial step in a peroxidase-based detection system. The detection of viral nucleic acid in formalin-fixed, paraffin-embedded sections using biotinylated and 35S-labelled probes is compared to the detection of viral antigen by immunocytochemistry. Biotinylated probes detected more positive cells than 35S labelled probes and were more specific. Biotinylated probes detected more positive cells than immunocytochemistry. PMID- 2547815 TI - Enterovirus IgM detection: specificity of mu-antibody-capture radioimmunoassays using virions and procapsids of Coxsackie B virus. AB - A predominantly type-specific mu-capture radioimmunoassay (RIA) of IgM antibodies to Coxsackie B1-B5 (CB1-CB5) viruses was previously described (Frisk et al., 1984). The present study is concerned with the specificity of this assay, using as antigen different strains of one serotype (CB5) and procapsids of two serotypes (CB3 and CB5). Eight strains of CB5 virions were tested against acute and/or convalescent sera from 10 patients from whom CB5 had been isolated. Seven patients' sera were tested against their own strain. The frequency of IgM positive patients varied from 9 of 10 (90%) to 5 of 10 (50%). In three cases the highest titres were obtained with the patients' own strain. When sera from patients with other enterovirus infections were tested against the CB5 strains, heterotypic titres were obtained to a certain extent (0-15.6%). The strains giving a high frequency of homotypic titres varied concerning heterotypic reactions. It is concluded that the choice of strain is important if a high frequency of homotypic titres with no or only a few heterotypic reactions is to be obtained. When procapsids were used as antigen, both homotypic and heterotypic titres were seen to a large extent. All patients with homotypic IgM against CB3 or CB5 virions showed IgM against the CB3 or the CB5 procapsids, respectively. When sera from patients with other enterovirus infections were tested, IgM was found in 54 of 93 patients (58%) with use of the CB3 procapsid and in 52 of 87 patients (60%) with the CB5 procapsid. It was often not the same patients who showed IgM against the two different procapsids. When both procapsids were used, IgM positivity was found in 62 of 81 patients (77%) with other enterovirus infections. It is concluded that the use of two or more procapsids in combination is of value for the diagnosis of a recent or current enterovirus infection. PMID- 2547816 TI - A capture antibody ELISA for detection of antibodies against bovine enterovirus. AB - A capture antibody ELISA was developed for detecting antibodies against bovine enterovirus. Chicken IgG F(ab')2 fragments against strain J2129 captured virus effectively. The captured virus was used to detect specific antibodies against the virus in bovine sera. The specificity of the ELISA is high. The factors affecting the specificity are discussed. The assay is more sensitive and economic than traditional serum neutralisation. PMID- 2547817 TI - Development of monoclonal neutralizing antibodies against bovine viral diarrhea virus after pretreatment of mice with normal bovine cells and cyclophosphamide. AB - After immunization of mice with bovine viral diarrhea virus (BVDV) concentrated by ultracentrifugation only antibodies against normal cellular constituents were detectable. Monoclonal and polyclonal anti-BVDV antibodies were obtained, however, after induction of tolerance against normal bovine cellular antigens by pretreatment of adult mice with bovine cells and cyclophosphamide followed by immunization with BVDV containing material. PMID- 2547818 TI - Rapid detection of human rotavirus strains in stools by single-sandwich enzyme linked immunosorbent assay systems using monoclonal antibodies. AB - Using murine monoclonal antibodies (MAbs) raised against the common antigen of group A rotavirus (RV), two single-sandwich ELISA systems were developed for detection of RV in stools: one using polyclonal antibody (PAb) as capture and a MAb as detector antibody (referred to as PAb-MAb assay); and the other based on the use of two different MAbs as capture and detector antibodies (referred to as MAb-MAb assay). In each single-sandwich ELISA system, samples and peroxidase labeled MAb were incubated sequentially (two-step method) or simultaneously (one step method). Using the two-step procedure on purified RV, 50 pg of protein was detected in the PAb-MAb as well as in the MAb-MAb assay, whereas the one-step method detected 0.4 ng and a conventional double-sandwich ELISA detected 3.2 ng of viral protein. Titration of RV samples from stools and cell cultures showed that single-sandwich ELISA titers were, on the average, 10-100-fold higher than those obtained by electron microscopy (EM), but 10-100-fold lower than those obtained by solid-phase immune EM (SPIEM). However, when 200 stool samples previously examined by EM or SPIEM were tested by the single-sandwich ELISA systems, specificity and sensitivity of these assays were 100%, and comparable to SPIEM. No false positive results were obtained when 54 samples of meconium and 91 stools from newborns in the first five days of life were tested. The two-step procedure appeared to be somewhat preferable over the one-step method, which, although faster, gave a marked prozone with a few samples in the MAb-MAb assay. The use of MAbs in rapid single-sandwich ELISA systems for RV detection in stools appears highly convenient, due to reliable results and short test performance times. PMID- 2547820 TI - Concentration and purification of feline leukaemia virus (FeLV) and its outer envelope protein gp70 by aqueous two-phase systems. AB - The major protective antigens of retroviruses are considered to be their glycosylated envelope proteins. However, the methods commonly employed to enrich and purify virus from culture media such as pelleting and density-gradient centrifugation result in a low recovery of the viral external glycoproteins. This is an obvious drawback when the virus is intended for use in a vaccine. In search for alternative methods to concentrate and purify FeLV, we have attempted extraction in two-phase systems based on water-soluble polymers (Albertsson PA., Biochem Biophys Acta 1958; 27: 378-395). A variety of polymer systems was tested. Some of them seem attractive for a large-scale concentration of the virus and/or its glycoprotein. The distribution between the phases of two FeLV proteins, the outer envelope protein, gp70, and the gag protein, p27, was determined. With a system composed of dextran sulfate and polyvinyl alcohol both the glycoprotein and the gag protein were almost completely recovered in the lower phase which constitutes about 3% of the total system in weight. The two proteins were more than 40-fold purified as calculated on protein basis. The proteins can be extracted readily. PMID- 2547821 TI - Identification of herpes simplex viruses by automated profile analysis of viral proteins. AB - In this report we describe a computerized virus identification system based upon the fact that viral proteins, separated by one-dimensional (ID) SDS-PAGE have unique banding patterns (protein profiles). Fifty-eight clinical strains of herpes simplex virus (HSV) were correctly identified by comparing their protein profiles with reference profiles of HSV, other viruses and uninfected cell cultures stored in a computer data base. PMID- 2547819 TI - Development and evaluation of an IgM capture enzyme immunoassay for diagnosis of recent Norwalk virus infection. AB - A capture enzyme immunoassay specific for Norwalk IgM class antibody was developed. The assay was moderately sensitive, identifying 33/53 (62%) of patients with naturally acquired Norwalk virus infection and 17/18 (94%) of experimentally infected volunteers. The assay was also specific for IgM class antibody and acute Norwalk virus infection and results were generally reproducible. A specific IgM response correlated with seroconversion by total antibody blocking assay and occurred independently of clinical symptoms. Among 81 symptomatic cases composing seven Norwalk outbreaks, specific IgM was absent from acute phase sera collected less than or equal to three days post onset, and was uncommon in sera collected within one week and after five weeks, with an optimal collection time at about two to three weeks. The Norwalk IgM capture immunoassay may be used to augment paired sera assays in the identification of Norwalk associated outbreaks of acute gastroenteritis. PMID- 2547822 TI - Bluetongue virus propagation and plaque assay: variation due to medium and serum supplement. AB - Two base media, minimal essential medium (MEM) and RPMI 1640, were supplemented with a variety of serum extenders and/or substitutes for the purpose of defining a medium formulation capable of supporting good cell (VERO) growth and virologic assays. Bluetongue virus (BTV), the prototype Orbivirus in the Reoviridae, was used in all studies. In general, VERO cells grown in RPMI performed better than those grown in MEM relative to cell growth, virus production and plaque assay. RPMI was better for supporting cell growth when serum extenders (NuSerum or SerXtend) were employed as supplements. Relative to virologic techniques, cells grown in RPMI produced higher virus titers in both propagation studies and plaque assays. The interval from infection to greater than 90% cytopathic effect (CPE) was consistently shorter with RPMI as the base medium. Cell cultures supported with RPMI base medium, supplemented with 3.5% FBS with SerXtend, provided the best overall performance relative to: (a) amount of virus produced by infected cell monolayers, (b) sensitivity to productive infection under overlay conditions (revealed the highest titer of a standard virus stock) and (c) plaque assay quality including cell quality, plaque size and plaque clarity. PMID- 2547823 TI - Inhibitory effects of various anticoagulants on the infectivity of human cytomegalovirus. AB - We assessed the extent to which the anticoagulants heparin, sodium citrate, Alsever's solution and ethylene diamine tetra-acetic acid (EDTA) (at standard low concentration or at high concentration as attained when only a small blood volume was collected into anticoagulant) inhibited the infectivity of human cytomegalovirus (HCMV). All four anticoagulants had minimal effects on cell associated virus, but heparin at either low or high concentration and EDTA at high concentration reduced the titre of cell-free virus by 10-100 fold. The large inhibitory effects of heparin and EDTA on cell-free virus indicated that sodium citrate or Alsever's solution were preferable as anticoagulants when blood was collected for assay of CMV viraemia by virus isolation. PMID- 2547824 TI - Rapid diagnosis of adenovirus pharyngoconjunctival fever: use of a monoclonal antibody-based ELISA test during an outbreak. AB - A monoclonal antibody-based solid-phase immunoassay (ELISA) was used to detect adenovirus antigen in clinical specimens during an outbreak of pharyngoconjunctival fever at a boarding school. The use of the kit enabled rapid diagnosis of the cause of the outbreak. The assay was also valuable in confirmation of tissue culture isolation of adenovirus. This kit could be of value to laboratories without tissue culture facilities and as an alternative to immunofluorescence for adenovirus confirmation in laboratories which have such facilities. PMID- 2547825 TI - An amplified dot immunoassay for the direct quantitation of adapted and wild strains of human cytomegalovirus. AB - A rapid and simple quantitative dot immunoassay for a cell-adapted reference strain of cytomegalovirus (CMV) and for wild strains of CMV present in clinical urine samples was developed. The assay was performed on nitrocellulose paper dotted with several dilutions of viral pellets free of cellular debris. Viral dilutions were treated with a monoclonal antibody to the major component of the viral capsid. To amplify the reaction, a three-dimensional complex of streptavidin and biotinylated horseradish peroxidase was used as the detector system. The dot immunoassay, which does not require cell cultures, yielded results within one day. A significant correlation was found between CMV titers obtained by dot immunoassay and CMV infectious units determined by immunoalkaline phosphatase staining of CMV-late antigen positive cells. PMID- 2547827 TI - 21-Hydroxylase deficiency in female hyperandrogenism: screening and diagnosis. AB - 21-Hydroxylase-deficient late-onset adrenal hyperplasia (LOAH) appears to affect 1-6% of hyperandrogenic women. Screening and diagnostic criteria for LOAH have not been well established, as these patients are clinically indistinguishable from other hyperandrogenic women. The following prospective study was undertaken to 1) determine the predictive value of screening hyperandrogenic women for LOAH with a morning follicular phase basal 17-hydroxyprogesterone (17-HP) level and 2) compare the various in vivo estimates of 21-hydroxylase activity after adrenal stimulation for the diagnosis of LOAH. Twenty-one euandrogenic control women (physically normal, without hirsutism, with regular menses, and a negative family history) were studied. The clinical population consisted of 164 consecutive unselected patients seen at the Division of Reproductive Endocrinology and Infertility of Johns Hopkins University School of Medicine between 1983 and 1987 demonstrating hirsutism and/or hyperandrogenic oligomenorrhea. Controls and patients underwent acute adrenal stimulation with 1 mg ACTH-(1-24), administered in the morning to fasting patients in the follicular phase of their menstrual cycle. Blood was sampled before and 30 min after ACTH-(1-24) administration. Steroid RIA determinations were performed for 17-HP, progesterone, testosterone, dehydroepiandrosterone sulfate, androstenedione, FSH, LH, and PRL. Three estimates of 21-hydroxylase activity were studied: the 17-HP level 30 min post ACTH (17-HP30), the change in 17-HP (delta 17-HP0-30) and the summed rate of change in 17-HP and progesterone ([delta 17-HP0-30) + delta P0-30]/30 min). The upper 95th percentiles for these estimates of 21-hydroxylase activity in control women were 9.6 nmol/L (316 ng/dL), 8.8 nmol/L (292 ng/dL), and 0.39 nmol/L.min (13 ng/dL.min), respectively. Thirteen of 164 (7.9%) hyperandrogenic women had at least 1 abnormal 21-hydroxylase measurement. Four of these women (2.4%) had 17-HP measurements 3- to 20-fold above the upper normal 95th percentile (17-HP30 greater than 36.3 nmol/L or 1200 ng/dL) and were considered as suffering from LOAH. In our population the 3 measures of 21-hydroxylase studied clearly differentiated the LOAH women from all others, although a single 17-HP level 30 min post-ACTH was the simplest and most cost effective. Nine other hyperandrogenic women (5.5%) had at least 1 abnormal 21-hydroxylase measurement less than 3-fold the upper normal 95th percentile value and were designated as having mild 21-hydroxylase deficiency.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2547826 TI - Detection of cytomegalovirus DNA using probes labelled with digoxigenin. AB - Cloned HindIII restriction fragments of cytomegalovirus (CMV) DNA (strain AD169) were labelled with biotin, digoxigenin or 32P and used as probes to detect CMV DNA. Probes biotinylated by nick translation, random hexanucleotide priming (RHP) or "photobiotin" were able to detect 10-50 pg of homologous DNA. Probes labelled with digoxigenin or 32P by RHP detected 0.1 pg of homologous DNA, and 1-10 CMV infected fibroblasts. Comparison of digoxigenin- and 32P-labelled probes in a DNA hybridisation assay on 186 urine specimens demonstrated that these probes were of similar sensitivity, detecting CMV DNA in 40 and 41 specimens, respectively. Positive results were obtained using this hybridisation assay with 11 of 14 specimens (79%) yielding CMV by virus isolation, and with 35 other specimens obtained from patients with laboratory evidence of CMV infection or symptoms compatible with CMV disease. Thus digoxigenin-labelled probes may provide an assay that can detect CMV DNA in specimens yielding a negative result by virus isolation. PMID- 2547828 TI - Blunted adrenocorticotropin but normal beta-endorphin release after human corticotropin-releasing hormone administration in depression. AB - Since the discovery of CRH in 1981, several investigators have reported abnormalities of the hypothalamic-pituitary-adrenal (HPA) system in response to direct stimulation of the corticotroph cells in patients with psychiatric disorders. To further explore HPA system integrity in major depressive disorders, 13 drug-free patients and normal subjects matched for age, sex, ovarian status, and body weight received 100 micrograms synthetic human CRH as an iv bolus dose. Compared to that in the normal subjects, in the depressed patients a significant attenuation of the net ACTH release after CRH administration (772 +/- 597 vs. 263 +/- 286 pmol/min.L; P less than 0.02) was observed, while beta-endorphin and cortisol responses did not differ significantly between the groups. The magnitudes of ACTH and cortisol release were negatively correlated in the patient group only (r = -0.67; P less than 0.01). Thus, the blunted ACTH response to CRH in depression might be related to hypercortisolemia, while the implications of the apparent dissociation of ACTH and beta-endorphin after CRH administration still remain unclear. Our data support the hypothesis that the hyperactivity of the HPA system in depression most likely is a consequence of CRH hypersecretion, the origin of which may be explained by abnormal central glucocorticoid receptor or neurotransmitter regulation. PMID- 2547829 TI - Insulin-like growth factor-I: autocrine secretion by human thyroid follicular cells in primary culture. AB - Insulin-like growth factor-I (IGF-I) stimulates the growth of thyroid cells of different animal species. However, conflicting results have been reported as regards the function and mechanism of the action of IGF-I at the thyroid level. This study was designed to determine whether normal human thyroid cells have IGF I receptors and whether IGF-I could act on such cells through an autocrine mechanism. Human thyroid follicular cells in primary culture, not contaminated by fibroblasts, were used. They had specific and saturable binding sites for IGF-I, as revealed by radiolabeled binding method, and displayed an average receptor number of 2000/cell. Under the same experimental conditions, insulin receptors were not detectable. Human thyroid follicular cells secreted IGF-I into the culture medium, as assessed by a specific chemiluminescent immunoassay. The IGF-I secretory process was detectable for at least 12 days of culture, but a high degree of variability has been found among individual samples. Acid-gel chromatography demonstrated that IGF-I and a higher mol wt IGF-I, most likely IGF I bound to its binding protein, were secreted by human thyroid cells. TSH stimulated the secretion of the two molecules in normal human thyroid cells. The TSH effect on IGF-I secretion was concentration dependent between 0.1 nmol/L and 0.1 mumol/L. GH stimulated IGF-I synthesis by thyroid cells in a concentration range from 20-200 micrograms/mL. Since binding studies demonstrated the presence of IGF-I receptors on human thyroid cells, IGF-I probably regulates human thyroid function through an autocrine mechanism. PMID- 2547830 TI - Short loop adrenocorticotropin (ACTH) feedback after ACTH-(1-24) injection in man is an artifact of the immunoradiometric assay. AB - A recent report measured a decrease in plasma ACTH concentration by immunoradiometric assay (IRMA) during infusion of ACTH-(1-24) in humans. It was concluded that this decrease in ACTH concentration was due to short loop ACTH autoregulation. The present study demonstrates that the decrease in ACTH concentration measured by IRMA was due to an artifact of the IRMA. We injected 250 micrograms ACTH-(1-24), iv, into five normal male volunteers after overnight 2.5-g metyrapone administration. The ACTH concentration measured by IRMA decreased from 59.6 +/- 9.7 pmol/L before to 4.8 +/- 2.0 pmol/L 1 min after ACTH (1-24) injection. The ACTH concentration measured by IRMA increased thereafter in a mirror image of the decline in ACTH-(1-24) measured by RIA. Addition of ACTH-(1 24) to plasma in vitro resulted in a decrease in the ACTH concentration measured by IRMA which was of similar magnitude to that observed in vivo. ACTH-(1-24) infusion in vivo or addition to ACTH-(1-39)-containing plasma in vitro decreased ACTH-(1-39) measured by IRMA by binding to N- but not C-terminal antibody without forming a detectable sandwich complex. We conclude that although ACTH short loop feedback may exist, it cannot be detected after ACTH-(1-24) injection with the use of a two-site IRMA. PMID- 2547831 TI - Three myosin heavy chain isoforms in type 2 skeletal muscle fibres. AB - Mammalian skeletal muscles consist of three main fibre types, type 1,2A and 2B fibres, with different myosin heavy chain (MHC) composition. We have now identified another fibre type, called type 2X fibre, characterized by a specific MHC isoform. Type 2X fibres, which are widely distributed in rat skeletal muscles, can be distinguished from 2A and 2B fibres by histochemical ATPase activity and by their unique staining pattern with seven anti-MHC monoclonal antibodies. The existence of the 2X-MHC isoform was confirmed by immunoblotting analysis using muscles containing 2X fibres as a major component, such as the normal and hyperthyroid diaphragm, and the soleus muscle after high frequency chronic stimulation. 2X-MHC contains one determinant common to 2B-MHC and another common to all type 2-MHCs, but lacks epitopes specific for 2A- and 2B-MHCs, as well as an epitope present on all other MHCs. By SDS-polyacrylamide gel electrophoresis 2X-MHC shows a lower mobility compared to 2B-MHC and appears to comigrate with 2A-MHC. Muscles containing predominantly 2X-MHC display a velocity of shortening intermediate between that of slow muscles and that of fast muscles composed predominantly of 2B fibres. PMID- 2547832 TI - Biphasic forearm vascular responses to intraarterial arginine vasopressin. AB - Forearm vascular responses to arginine vasopressin (AVP) infused into a brachial artery in a wide range of infusion rates (0.05-2.0 ng/kg per min) were examined in 20 young healthy volunteers. Intraarterial AVP at lower doses (0.05 and 0.1 ng/kg per min) caused forearm vasoconstriction, whereas AVP at a dose of 0.2 ng/kg per min or higher caused forearm vasodilatation. The maximal forearm vasoconstriction was induced at the venous plasma AVP level of 76.3 +/- 8.8 pg/ml. Forearm vasodilatation was associated with the venous plasma AVP level of 369 +/- 43 pg/ml or higher. Forearm vasodilatation was the result of the direct effect of AVP since forearm blood flow and vascular resistance in the contralateral arm did not change. We attempted to explore the mechanisms involved in AVP-induced direct vasodilatation. The treatment with indomethacin, 75 mg/d for 3 d, did not alter AVP-induced forearm vasodilatation. In contrast, intraarterial infusion of isoosmolar CaCl2 totally prevented AVP-induced forearm vasodilatation. Intra-arterial CaCl2 also markedly attenuated forearm vasodilatation induced by intraarterial sodium nitroprusside, but did not alter forearm vasodilatation induced by intraarterial isoproterenol. These results indicate that the direct vascular effects of intra-arterial AVP on the forearm vessels are biphasic, causing vasoconstriction at lower doses and vasodilatation at higher doses. The direct vasodilatation induced by intraarterial AVP at higher doses is not mediated by prostaglandins but may involve cGMP-related mechanisms. PMID- 2547833 TI - T cell receptor gamma and delta rearrangements in hematologic malignancies. Relationship to lymphoid differentiation. AB - We have studied recombinatorial events of the T cell receptor delta and gamma chain genes in hematopoietic malignancies and related these to normal stages of lymphoid differentiation. T cell receptor delta gene recombinatorial events were found in 91% of acute T cell lymphoblastic leukemia, 68% of non-T, non-B lymphoid precursor acute lymphoblastic leukemia (ALL) and 80% of mixed lineage acute leukemias. Mature B-lineage leukemias and acute nonlymphocytic leukemias retained the T-cell receptor delta gene in the germline configuration. The incidence of T cell receptor gamma and delta was particularly high in CD10+CD19+ non-T, non-B lymphoid precursor ALL. In lymphoid precursor ALL, T cell receptor delta was frequently rearranged while T cell receptor gamma was in the germline configuration. This suggests that TCR delta rearrangements may precede TCR gamma rearrangements in lymphoid ontogeny. In T-ALL, only concordant T cell receptor delta and gamma rearrangements were observed. Several distinct rearrangements were defined using a panel of restriction enzymes. Most of the rearrangements observed in T-ALL represented joining events of J delta 1 to upstream regions. In contrast, the majority of rearrangements in lymphoid precursor ALL most likely represented D-D or V-D rearrangements, which have been found to be early recombinatorial events of the TCR delta locus. We next analyzed TCR delta rearrangements in five CD3+TCR gamma/delta+ ALL and cell lines. One T-ALL, which demonstrated a different staining pattern with monoclonal antibodies against the products of the TCR gamma/delta genes than the PEER cell line, rearranges J delta 1 to a currently unidentified variable region. PMID- 2547834 TI - Regulation by fasting of rat insulin-like growth factor I and its receptor. Effects on gene expression and binding. AB - We have examined, in liver and extrahepatic tissues, the effects of fasting on total insulin-like growth factor I (IGF-I) mRNA levels, on levels of different IGF-I mRNAs generated by alternative splicing of the primary IGF-I transcript, and on IGF-I receptor binding and mRNA levels. A 48-h fast decreased total IGF-I mRNA levels by approximately 80% in lung and liver, approximately 60% in kidney and muscle, and only approximately 30-40% in stomach, brain, and testes. In heart, IGF-I mRNA levels did not change. The levels of the different splicing variants, however, were essentially coordinately regulated within a given tissue. Specific 125I-IGF-I binding in lung, testes, stomach, kidney, and heart was increased by fasting by approximately 30-100%, whereas in brain 125I-IGF-I binding did not change in response to fasting. In tissues in which fasting increased IGF-I receptor number, receptor mRNA levels increased approximately 1.6 to 2.5-fold, whereas when IGF-I receptor number was unchanged in response to fasting, receptor mRNA levels did not change. These data demonstrate that the change in IGF-I and IGF-I receptor mRNA levels during fasting is quantitatively different in different tissues and suggest that regulation of IGF-I and IGF-I receptor gene expression by fasting is discoordinate. PMID- 2547835 TI - Endothelin-induced increases in vascular smooth muscle Ca2+ do not depend on dihydropyridine-sensitive Ca2+ channels. AB - Endothelin is a potent mammalian vasoconstrictive peptide with structural homology to cation channel-binding insect toxins. We tested the proposal that this peptide directly activates dihydropyridine-sensitive Ca2+ channels in cultured vascular smooth muscle (VSM) cells. First, we found that cell Ca2+ can be altered in VSM by activation of voltage-operated Ca2+ channels. KCl-induced depolarization and the dihydropyridine Ca2+ channel agonist (-) Bay K 8644 (10 microM) both raised cell Ca2+ more than twofold; the effect of KCl was blocked by the inhibitory enantiomer, (+) Bay K 8644 (40 microM). Similar responses were observed in Chinese hamster ovary (CHO) cells. Synthetic endothelin (4 x 10(-8) M) raised Ca2+ in VSM but not CHO cells from 100 +/- 17 to 561 +/- 34 nM within 12 s. Ca2+ subsequently fell to basal levels after 30 min. Half maximal Ca2+ response was at 4 x 10(-9) M endothelin. Unlike endothelin, thrombin raised Ca2+ in both VSM and CHO cells. The Ca2+ responses to endothelin and thrombin were not affected by nicardipine (1 microM), (+) Bay K 8644, or Ca2+-free solutions. Lastly, both hormones caused release of inositol phosphates in VSM cells. However, the response to thrombin was more than 10-fold larger and was more rapid than the response to endothelin; the thrombin response was sensitive to pertussis toxin, while the response to endothelin was not. Thus endothelin, like thrombin, raises cell Ca2+ in VSM by mobilization of intracellular stores and not by activation of dihydropyridine-sensitive Ca2+ channels. However, their receptors are distinct and they exhibit important differences in signal transduction. PMID- 2547837 TI - Endothelin binding to cultured calf adrenal zona glomerulosa cells and stimulation of aldosterone secretion. AB - Endothelins are a group of potent vasoconstrictors whose structure was deduced from genomic DNA. ET-1 was first isolated from culture supernatants from porcine endothelial cells and ET-3 was identified from a rat DNA library. We report on the binding of 125I-ET-1 to zona glomerulosa cells in culture and on its ability to stimulate aldosterone secretion. Cultured calf adrenal zona glomerulosa cells have saturable, high affinity [Kd = 1.00 +/- 0.17 X 10(-10) M (SEM)] receptors which bind ET-1 in a temperature and time dependent manner. Binding was specific and angiotensin II, vasopressin, ANP, BNP, apamin, calcium channel agonists or antagonists did not interact with the receptor. ET-3 displaced 125I-ET-1 from the receptor with a relative potency of 0.39 +/- 0.1% (SEM) that of ET-1. ET-1 incubated with cultured glomerulosa cells stimulated aldosterone secretion in a dose dependent manner but it was less potent than angiotensin II. ET-3 had less than 1% the relative potency of ET-1 stimulating aldosterone secretion. This data suggest that ET-1 is an independent stimulator of aldosterone secretion and we are speculating that it might be important in those situations, like in malignant hypertension, where endothelial damage might result in increased ET-1 production. PMID- 2547836 TI - Human osteoblasts in vitro secrete tissue inhibitor of metalloproteinases and gelatinase but not interstitial collagenase as major cellular products. AB - Human osteoblast cultures (hOB) were examined for the production of interstitial collagenase, tissue inhibitor of metalloproteinases (TIMP), and gelatinolytic enzymes. Cells were isolated by bacterial collagenase digestion of trabecular bone (vertebra, rib, tibia, and femur) from 11 subjects (neonatal to adult). Confluent cultures were exposed to phorbol 12-myristate 13-acetate, PTH, PGE2, epidermal growth factor, 1,25(OH)2 vitamin D3, recombinant human IL-1 beta, and dexamethasone. Collagenase and TIMP were assayed immunologically and also by measurements of functional activity. Collagenase was not secreted in significant quantities by human bone cells under any tested condition. Furthermore, collagenase mRNA could not be detected in hOB. However, hOB spontaneously secreted large amounts of TIMP for at least 72 h in culture. hOB TIMP was found to be identical to human fibroblast TIMP by double immunodiffusion, metabolic labeling and immunoprecipitation, Northern blot analysis, and stoichiometry of collagenase inhibition. SDS-substrate gel electrophoresis of hOB-conditioned media revealed a prominent band of gelatinolytic activity at 68 kD, and specific polyclonal antisera established its identity with the major gelatinolytic protease of human fibroblasts. Abundant secretion of gelatinolytic, but not collagenolytic, enzymes by hOB may indicate that human osteoblasts do not initiate and direct the cleavage of osteoid collagen on the bone surface, but may participate in the preparation of the bone surface for osteoclast attachment by removal of denatured collagen peptides. The constitutive secretion of TIMP may function to regulate metalloproteinase activity. PMID- 2547838 TI - Extracellular protons acidify osteoclasts, reduce cytosolic calcium, and promote expression of cell-matrix attachment structures. AB - Because metabolic acids stimulate bone resorption in vitro and in vivo, we focused on the cellular events produced by acidosis that might be associated with stimulation of bone remodeling. To this end, we exposed isolated chicken osteoclasts to a metabolic (butyric) acid and observed a fall in both intracellular pH and cytosolic calcium [( Ca2+]i). These phenomena were recapitulated when bone resorptive cells, alkalinized by HCO3 loading, were transferred to a bicarbonate-free environment. The acid-induced decline in osteoclast [Ca2+]i was blocked by either NaCN or Na3VO4, in a Na+-independent fashion, despite the failure of each inhibitor to alter stimulated intracellular acidification. Moreover, K+-induced membrane depolarization also reduced cytosolic calcium in a manner additive to the effect of protons. These findings suggest that osteoclasts adherent to bone lack functional voltage-operated Ca2+ channels, and they reduced [Ca2+]i in response to protons via a membrane residing Ca-ATPase. Most importantly, acidosis enhances formation of podosomes, the contact areas of the osteoclast clear zone, indicating increased adhesion to substrate, an early step in bone resorption. Thus, extracellular acidification of osteoclasts leads to decrements in intracellular pH and calcium, and appears to promote cell-matrix attachment. PMID- 2547839 TI - Platelet receptor-mediated factor X activation by factor IXa. High-affinity factor IXa receptors induced by factor VIII are deficient on platelets in Scott syndrome. AB - We have studied factor IXa binding and factor X activation with normal platelets and with platelets obtained from a patient with a bleeding disorder and an isolated deficiency of platelet procoagulant activity termed Scott syndrome. In the absence of factor VIIIa and factor X, normal, thrombin-treated platelets exposed 560 +/- 35 sites for factor IXa with a Kd of 2.75 +/- 0.27 mM, compared with 461 +/- 60 sites per patient platelet with Kd of 3.2 +/- 0.33 nM. The addition of factor VIIIa and factor X resulted in a decrease in the Kd for normal platelets to 0.68 nM but had no effect on the Kd for patient platelets. The concentrations of factor IXa required for half-maximal rates of factor X activation for normal (0.52 nM) and patient platelets (2.5 nM) were similar to those determined from equilibrium binding studies. Kinetic parameters for factor X activation by factor IXa showed that the Km and Kcat were identical for normal and patient platelets in the absence of factor VIIIa. In the presence of factor VIIIa, and kcat for patient platelets (163 min-1) was only 33% of that for normal platelets (491 min-1): This result can be explained by the difference in affinity for factor IXa between normal and patient platelets in the presence of factor VIIIa, suggesting impaired factor VIIIa binding to Scott syndrome platelets. PMID- 2547841 TI - Cl- secretion induced by bile salts. A study of the mechanism of action based on a cultured colonic epithelial cell line. AB - When applied to the basolateral (serosal) side of the T84 colonic epithelial monolayer, taurodeoxycholate caused net Cl- secretion in a dose-dependent manner with a threshold effect observed at 0.2 mM. In contrast, when applied to the apical (luminal) surface, concentrations of taurodeoxycholate below 1 mM had little or no effect. Only when the concentration of taurodeoxycholate present on the apical side was greater than or equal to 1 mM did apical addition results in an electrolyte transport effect. This apical effect on electrolyte transport was associated with an abrupt increase in the permeability of the monolayer. Cyclic AMP and cyclic GMP in the T84 monolayers were not increased by the bile salt, but in the presence of extracellular Ca2+, free cytosolic Ca2+ increased with a graded dose effect and time course that corresponded approximately to the changes in short circuit current (Isc). The results suggest that luminal bile salts at a relatively high concentration (greater than or equal to 1 mM) increase tight junction permeability. Once tight junction permeability increases, luminal bile salts could reach the basolateral membrane of the epithelial cells where they act to increase free cytosolic Ca2+ from extracellular sources. The resulting increases in free cytosolic Ca2+, rather than in cyclic nucleotides, appear to be involved in transcellular Cl- secretion. PMID- 2547840 TI - Autonomous growth of a human neuroblastoma cell line is mediated by insulin-like growth factor II. AB - Insulin-like growth factor II (IGF-II) mRNA was increased in two of eight neuroblastomas and in eight of eight pheochromocytomas, tumors of the adrenal medulla that occur in childhood and adulthood, respectively. RNA encoding the type I IGF receptor, the receptor thought to mediate the mitogenic effects of IGF I and IGF-II, also was uniformly expressed in these cells. To assess the role of IGF-II in the growth of these tumor cells, we have used the SK-N-AS cultured neuroblastoma cell line, which can be continuously propagated in mitogen-free medium, as a model system. Our results strongly suggest that IGF-II, synthesized by SK-N-AS cells and acting through type I IGF receptors, contributes to the autonomous growth of this tumor cell line. (a) SK-N-AS cells synthesized large amounts of IGF-II RNA and secreted greater than 50 ng/ml of IGF-II (as determined by specific radioimmuno- and radioreceptor assays). Little, if any, IGF-I RNA or immunoreactive IGF-I were detected. (b) SK-N-AS cells possess type I IGF receptors. (c) Exogenous IGF-I and IGF-II stimulated DNA synthesis in SK-N-AS cells, and this stimulation was abolished by a blocking antibody to the type I IGF receptor. (d) This anti-receptor antibody also abolished the multiplication of SK-N-AS cells in the absence of added mitogens. We conclude that IGF-II is an autocrine growth factor for SK-N-AS cells and suggest that this mechanism may contribute to the growth of some adrenal medullary tumors. PMID- 2547842 TI - Hepatic phosphotyrosine phosphatase activity and its alterations in diabetic rats. AB - Phosphotyrosine phosphatase (PTPase) activity in rat liver was measured using a phosphopeptide substrate containing sequence identity to the major site of insulin receptor autophosphorylation. PTPase activity was detected in both cytosolic and particulate fractions of rat liver and produced linear dephosphorylation over a 15-min time course. In rats made insulin-deficient diabetic by streptozotocin treatment (STZ), cytosolic PTPase activity increased to 180% of the control values after 2 d of diabetes and remained elevated at 30 d (P less than 0.02). Gel filtration on Sephadex-75 revealed a single peak of activity in the cytosol in both control and diabetic animals and confirmed the increased levels. In BB diabetic rats, another model of insulin deficiency, the PTPase activity in the cytosolic fraction was increased to approximately 230% of control values. PTPase activity in the particulate fraction of liver was also increased by 30 and 80% after 2 and 8 d of STZ diabetes, respectively. However, this increase was not sustained and after 30 d of STZ diabetes, PTPase activity associated with the particulate fraction in the BB diabetic rat was reduced to approximately 70% of the control levels. Treatment of STZ diabetic rats with subcutaneous insulin or vanadate in their drinking water for 3 d reduced tyrosine PTPase activity in the particulate, but not in the cytosolic fraction. This was associated with a change in blood glucose toward normal. These data indicate insulin deficient diabetes is accompanied by significant changes in hepatic PTPase activity. Since tyrosine phosphorylation plays a central role in the cellular action of insulin receptor, an increase in PTPase activity may be an important factor in the altered insulin action associated with these diabetic states. PMID- 2547843 TI - Regulation of rabbit medullary collecting duct cell pH by basolateral Na+/H+ and Cl-/base exchange. AB - The collecting duct of the inner stripe outer medulla (OMCDi) is a major site of distal nephron acidification. Using the pH sensitive fluorescent dye 2'-7' bis(carboxyethyl)-5,6,-carboxyfluorescein (BCECF) and quantitative spectrofluorometry to measure intracellular pH in isolated perfused OMCDi, we have characterized basolateral transport processes responsible for regulation of intracellular pH. Experiments suggesting the existence of basolateral Cl-/base exchange were performed. In HCO3- containing buffers, bath Cl- replacement resulted in reversible alkalinization of the OMCDi from 7.22 +/- 0.05 to 7.57 +/- 0.12. Similarly 0.1 mM bath 4',4'-diisothiocyanostilbene-2,2'-disulphonic acid (DIDS) alkalinized the OMCDi from 7.14 +/- 0.09 to 7.34 +/- 0.09 and blocked further alkalinization by bath Cl- removal (delta = + 0.02 pH units). The concentration dependence kinetics of Cl-/base exchange revealed a K1/2 of 10 mM for external Cl- with a Vmax of 0.50 pH U/min. Experiments suggesting the existence of basolateral Na+/H+ exchange were also performed. Replacement of bath Na+ by tetramethylammonium resulted in reversible cell acidification (7.14 +/- 0.09 to 6.85 +/- 0.1). Tubules that were acidified by a brief exposure to NH4Cl displayed recovery of cell pH back to baseline at a rate that was highly dependent on bath Na+ concentration. Half maximal recovery rate was achieved at 7 mM bath Na+ and Vmax was 0.605 pH U/min. The Na+-dependent rate of cell pH recovery after acidification was blocked by 0.2 mM bath amiloride. These results suggest that intracellular pH in the OMCDi is regulated by parallel basolateral Na+/H+ exchange and Cl-/base exchange. PMID- 2547844 TI - Laryngeal papillomatosis: correlation between severity of disease and presence of HPV 6 and 11 detected by in situ DNA hybridisation. AB - A technique using a biotin-streptavidin polyalkaline phosphatase complex was applied to routinely fixed and processed biopsy specimens of laryngeal papillomata from 45 patients taken over the past 20 years to detect human papilloma virus (HPV) types 6 and 11. Two thirds of both adult and juvenile onset cases were positive for HPV 6 or HPV 11 or both. Five specimens of normal vocal cord epithelium were negative for HPV 6 and 11. The detailed clinical history, endoscopic findings, success of treatment and eventual prognosis were compared with the HPV state of biopsy material for each patient. Patients with multiple confluent lesions when first seen, whose histology showed florid koilocytosis and who had strongly positive reactivity for HPV 6 or 11 present in the surface epithelial cell nuclei, had a poor prognosis requiring multiple endoscopies to control their disease. PMID- 2547846 TI - Influence of cancer histology on the success of fine needle aspiration of the breast. AB - Fine needle aspiration (FNA) cytology carried out on 1318 primary breast cancers from 1980 to 1986 inclusive showed that 198 were well recognised, histological special types. These included medullary, mucoid, tubular, cribriform and lobular invasive cancers, and non-invasive cancers. Excluding these special histological types, the overall number successfully identified (malignant plus suspicious) by fine needle aspiration was 940 (84%), although in only 820 (73%) was malignancy definitely diagnosed. The results for the special types were variable, the mucoid and medullary cancers being consistently identified while the other types were not. For tubular and cribriform, lobular and non-invasive ductal cancers a malignant diagnosis was made in 30% to 40% of cases, although inclusion of suspicious results gave identification figures of 60% to 70%. Particular cytological patterns are characteristic of some special histological cancer types but lesion cellularity, size, and physical definition are all intrinsic factors influencing success of FNA diagnosis. About 10% of all primary breast cancers seem to have certain histological properties which further reduce the success of fine needle aspiration. PMID- 2547845 TI - Quantity of nuclear DNA in malignancies and benign lymphadenopathies associated with Epstein-Barr virus. AB - The quantity of nuclear DNA in 90 tumours with a strong probability of being associated with Epstein-Barr virus (EBV) (11 cases with nasopharyngeal carcinomas, seven cases of endemic and 26 of non-endemic Burkitt's lymphoma, and 46 cases of Hodgkin's disease) were analysed by flow cytometry. Twenty three cases with benign lymphadenopathies were analysed in a similar way. Except for endemic Burkitt's lymphoma most of the tumours were diploid. Near-diploid aneuploidy (with a DNA index ranging from 1.06 to 1.26) was also found in endemic Burkitt's lymphoma as well as in six non-endemic Burkitt's lymphomas and in eight cases of Hodgkin's disease but was absent in nasopharyngeal carcinomas. Tetraploid aneuploidy was seen in three cases of nasopharyngeal carcinoma. Five of the 23 cases of lymphadenopathy also showed near-diploid DNA, one of which subsequently developed into a non-Hodgkin's lymphoma. It is concluded that near diploid DNA content seems to be associated with the lymphatic origins of the tumours rather than with EBV. PMID- 2547847 TI - Detection of cytomegalovirus antigens and DNA in tissues fixed in formaldehyde. AB - Immunohistochemical techniques with monoclonal antibodies against cytomegalovirus (CMV) immediate early (IEA) and early antigens (EA), and in situ hybridisation, were used to detect CMV infection in routinely obtained, formaldehyde fixed and paraffin wax embedded tissues taken from bone marrow transplant patients, who had died form interstitial pneumonia. To improve the rates of detection of CMV-IEA and EA the wax embedded material was pretreated with 0.4% pepsin/HCl at 37 degrees C for 30 minutes. This pretreatment was also advantageous for in situ hybridisation. In the patients with histological evidence of CMV infection or positive viral culture from the lung tissue, or both, viral proteins and nucleic acids were detected in lung, as well as in other organs. Immunohistochemical techniques proved superior in heavily infected but necrotic tissues. In control patients (patients who had died from interstitial pneumonia without any evidence of CMV, or with no interstitial pneumonia at all) in situ hybridisation showed no positive signal, while immunohistochemical techniques showed only a few positive cells in lung tissue of one of nine patients. In addition to CMV-DNA analysis, formaldehyde-fixed, paraffin wax embedded tissue is amenable to immunohistochemical analysis with CMV monoclonal antibodies. PMID- 2547848 TI - A dose-response evaluation of pipecuronium bromide in elderly patients under balanced anesthesia. AB - Pipecuronium bromide is a new steroidal non-depolarizing muscle relaxant currently under investigation. It is similar to pancuronium with respect to the duration of action, but lacking its cardiovascular side effects. We examined the dose-response relation of pipecuronium in 27 patients, ages 66-79 years, utilizing the incremental dose method under balanced anesthesia. The ED50, ED90 and ED95 were 22.42 (5.2) mcg/kg, 31.81 (6.9) mcg/kg and 35.12 (7.8) mcg/kg, respectively (log probit method). Our recovery data also demonstrate that residual neuromuscular blockade due to pipecuronium can easily be antagonized with neostigmine as long as spontaneous recovery of T1- at the time of reversal administration is greater than 13%. The authors conclude that under balanced anesthesia the cumulative dose-response of pipecuronium in the elderly patients is consistent with those previously described for younger population. Therefore, no dose adjustment appears necessary for the elderly. However, as with all medications, careful administration is appropriate. PMID- 2547849 TI - Nerve growth factor receptor immunoreactive profiles in the normal, aged human basal forebrain: colocalization with cholinergic neurons. AB - A monoclonal antibody raised against the receptor for nerve growth factor (NGF) has been used to map the distribution of NGF receptor-containing profiles within the human basal forebrain of four male and three female elderly patients without neurologic or psychiatric illness. Immunohistochemically processed tissue reveals a continuum of NGF receptor-positive neurons located within the medial septum, vertical and horizontal limb nuclei of the diagonal band, and nucleus basalis. NGF receptor-containing neurons are also found within the bed nucleus of the stria terminalis, the anterior commissure, the internal capsule, and the internal and external medullary laminae of the globus pallidus. Virtually all (greater than 95%) NGF receptor-containing neurons colocalize with the specific cholinergic marker choline acetyltransferase (ChAT) or the nonspecific marker acetylcholinesterase (AChE). Conversely, a few cholinergic perikarya are found which are not NGF receptor positive (and vice versa). These findings demonstrate that human basal forebrain neurons on which NGF receptor immunoreactivity is detected are primarily cholinergic and analogous to the nonhuman primate Ch1-Ch4 subgroups of Mesulam et al. (J. Comp. Neurol., 214:170-197, '83). NGF receptor containing fiber tracts are observed emanating from the medial septum and vertical limb nucleus of the diagonal band coursing medially within the fornix. Another fascicle originating mainly from the nucleus basalis and travelling within the external capsule enroute to the cortex is observed innervating all cortical layers. Comparison of NGF receptor- and ChAT-containing neurons reveals cholinergic perikarya within the striatal complex, whereas virtually no NGF receptor-containing neurons are found in these structures. An occasional displaced NGF receptor-containing neurons is seen in the ventrolateral portion of the putamen and the white matter underlying the nucleus accumbens. These data are discussed in terms of the relationship of NGF receptor- and ChAT-containing neurons within the basal forebrain and in terms of the possible functional significance of NGF in normal and diseased brain. PMID- 2547850 TI - Spatial organization of thalamocortical and corticothalamic projection systems in the rat SmI barrel cortex. AB - Axonal tracing techniques were used to examine the distribution of corticothalamic projection neurons in relation to the organization of the thalamocortical recipient zones in the whisker representation of the rat first somatic sensory cortex. Following injection of horseradish peroxidase into the physiologically defined vibrissa area in the ventrobasal complex of the thalamus, labeling in the cortex had a columnar appearance. Dense patches of anterograde labeling were located within the centers of the layer IV barrels and extended superficially through lamina III; the septa between barrels contained considerably less reaction product. Retrogradely labeled neurons were observed in lower layer V and layer VI where they were concentrated preferentially deep to the barrel centers. Regions deep to the septa displayed less overall labeling and a lower relative number of thalamic projecting neurons. Zones having the larger numbers of retrogradely labeled cells also contained terminallike labeling of either corticothalamic or thalamocortical origin. Following an injection that included the posterior group medial to the ventrobasal complex, anterograde labeling in layer IV was located largely in the septa. In conjunction with previous findings concerning the origin and termination of other projection systems in the barrel cortex, these results suggest that a vibrissal column contains a central core zone intimately linked with the ventrobasal thalamus that is bounded by narrower regions of more diverse inputs and outputs that form an interface between adjacent cortical columns. PMID- 2547851 TI - Chemoanatomical organization of the noradrenergic input from locus coeruleus to the olfactory bulb of the adult rat. AB - The locus coeruleus contains noradrenergic neurons which project widely throughout the CNS. A major target of locus coeruleus projections in the rat is the olfactory bulb (Shipley et al.: Brain Res. 329:294-299, '85) but the organization of the projections within the bulb has not been systematically examined. In this study, the laminar distribution and densities of locus coeruleus-noradrenergic fibers in the main and accessory olfactory bulbs were determined with anterograde tracing and immunocytochemical techniques. Following iontophoretic injections of 1% wheat germ agglutinin-horseradish peroxidase into the locus coeruleus, the densest anterograde label in the accessory olfactory bulb was observed in the external plexiform layer, granule cell layer, and especially in the internal part of the mitral cell layer. Virtually no label was observed in the glomerular layer. In the main olfactory bulb, labelled axons were observed in the granule cell layer, in the internal and external plexiform layers, occasionally in the mitral cell layer, and least often in the glomerular layer. Noradrenergic fibers in the olfactory bulb were identified by using immunocytochemistry with an antibody to dopamine-beta-hydroxylase. Laminar patterns and densities of noradrenergic innervation were determined with quantitative image analysis. In the accessory olfactory bulb, the densest innervation was in the innermost portion of the mitral cell layer followed by the granule cell layer, the superficial part of the mitral cell layer, and the external plexiform layer. The density of fibers in the glomerular layer was least. The laminar pattern of noradrenergic fiber distribution in the main olfactory bulb was similar to that in accessory olfactory bulb. The present studies demonstrate that locus coeruleus-noradrenergic fibers terminate preferentially in the internal plexiform, granule cell, and external plexiform layers. This suggests that the major influence of the locus coeruleus input to both the main and accessory the olfactory bulbs is on the predominant neuronal element in those layers, the granule cells. Additional studies are needed to resolve how this input influences specific olfactory bulb circuits. PMID- 2547852 TI - Verifying copper toxicosis in Bedlington terriers by analytical electron microscopy of needle biopsies of liver. AB - Diagnosis of copper toxicosis in Bedlington terriers was performed by analytical electron microscopy of needle biopsy of the liver. The animals were four Bedlington terriers clinically suspected of hereditary copper toxicosis, and four healthy control animals of the same breed. Light microscopy of the livers of the diseased animals revealed dense bodies in the hepatocytes which stained histochemically for copper. Analytical electron microscopy showed that the intrahepatocytic bodies consisted almost entirely of copper. PMID- 2547853 TI - Tubuloreticular inclusions in equine connective tissue neoplasms. AB - Abnormal irregularly branched and anastomosing tubules within cisternae of endoplasmic reticulum were observed by transmission electron microscopy in tumour cells comprising connective tissue neoplasms (sarcoids) from three horses and a mule. These tubuloreticular inclusions were also observed in cultured tumour cells from one of these horses examined, but were not detected in fibroblasts (fibrocytes), epidermis, or vascular endothelial cells in skin biopsy specimens from five clinically healthy horses, nor in one additional equine connective tissue neoplasm. PMID- 2547854 TI - Comparison of human papillomavirus type 1 serotyping by monoclonal antibodies with genotyping by in situ hybridization of plantar warts. AB - Thirty plantar warts were analyzed for the presence of HPV-1 type-specific and PV genus-specific capsid antigens by immunofluorescence (IF) using monoclonal and polyclonal antibodies and type-specific HPV-1 DNA employing in situ hybridization methods. Fifteen of 30 plantar warts were positive by IF for PV genus-specific structural viral antigens. Thirteen of the 15 productively infected plantar warts expressed intranuclear HPV-1 type-specific capsid antigens and viral DNA, which were detected in the same distribution in each individual wart. The 2 productively infected plantar warts that did not react with HPV-1 type-specific MoAbs did not react with HPV-1 type-specific DNA by in situ hybridization. Thus, serotyping of HPV-1 capsid antigens by monoclonal antibodies is concordant with genotyping of HPV-1 viral DNA by in situ hybridization in productively infected plantar warts. PMID- 2547855 TI - Phosphorylation of casein components by tyrosine-specific protein kinases. AB - The phosphorylation of different caseins by a number of tyrosine-specific kinases has been studied. The different kinases showed a range of specificities, suggesting differences in recognition mechanisms. The site of phosphorylation of alpha s2-casein by one phosphorylase has been identified, and compared with analogous sites in other proteins. PMID- 2547856 TI - Dental abnormalities and bone lesions associated with familial adenomatous polyposis: report of cases. AB - Dental abnormalities and bone lesions of the maxilla and mandible are found in more than 70% of patients with familial adenomatous polyposis (FAP). Dental abnormalities include impacted teeth (other than third molars), supernumerary teeth, congenitally missing teeth, and fused roots of molars. Bone lesions consist of osteomas--isolated or in clusters--in the maxilla and mandible. In this study of 24 patients, 75% had demonstrable dental abnormalities, and 79% had osteomas of the maxilla and mandible. Because dental abnormalities and bone lesions are present early in life, these features may be helpful in diagnosing this syndrome. PMID- 2547857 TI - Cross-reactivity of IgE antibodies to caddis fly with arthropoda and mollusca. AB - We investigated the possibility that subjects with IgE antibodies to an inhalant insect allergen, such as caddis fly, might also have antibodies to cross-reacting carbohydrate determinants (CCDs). IgE antibodies to cross-reacting allergens in caddis flies, mussels, oysters, shrimps, crabs, honeybee, and yellow jacket venoms were determined by RAST, RAST inhibition, and immunoblot studies with sera from three different sources: (1) sera of patients with well-defined inhalant atopy to caddis fly, (2) sera with IgE anti-CCD antibodies from subjects without known exposure to caddis fly, and (3) hyperimmune antisera with IgG anti-CCD antibodies raised as a result of immunization of rabbits with grass-pollen extract, buckwheat glycoprotein, or with honeybee venom. Sera from groups 2 and 3 reacted with Sepharose-coupled caddis fly extract in a RAST-type assay and elicited virtually identical patterns on immunoblots of caddis fly extract separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, whereas the sera from group 1 atopic patients did not react with CCD-rich material. However, indications for other types of cross-reacting antibodies were detected. The IgE antibodies of one of the patients studied (who was allergic not only to caddis fly but also to shellfish) were found to detect a cross-reacting homologous protein in extracts of mussel, oyster, shrimp, crab, honeybee, and yellow jacket venom. Preliminary results suggest that this cross-reacting 13 kd protein, the most prominent caddis fly allergen, is an invertebrate hemoglobin (erythrocruorin)-like molecule. These studies suggest the possibility that patients sensitized by exposure to caddis fly antigens could develop allergic reactions during their first exposure to shellfish or to their first bee sting. PMID- 2547859 TI - Frequency of food use data and the multidimensionality of diet. AB - Cancer epidemiologists rely heavily on the frequency of food use technique to assess dietary risk factors. We found that the accepted procedures for arraying individuals along a continuum, based on their positions within distributions of intakes of specific nutrients, simultaneously array them on other dietary characteristics. A unidimensional approach to dietary assessment could confound cancer risk assessments, and the effects could differ for men and women. We found that men consumed more calories and energy-containing nutrients than women, who were more likely to consume larger amounts of vitamins A and C. Dietary variety was similar for men and women, despite compositional differences in diet. Diets of men contained proportionately more meats, grains and nuts, and alcohol. Diets of women contained more fruits, vegetables, and poultry and fish. Among men, we found stronger intercorrelations among energy intake and intakes of fat, fiber, and vitamin C and among fat intake and intakes of fiber and vitamins A and C. Compositional differences in diet across quartiles of nutrient intake were not consistent for men and women, suggesting that the constellation of dietary risk factors may differ for the two sexes. Men in the lowest quartiles of energy, fiber, and vitamin A intakes had the greatest percent contribution of alcohol to the diet. This pattern was not observed for fat intake levels. Measures of dietary patterns may be needed for more accurate descriptions of the associations of cancer risk and diet. PMID- 2547858 TI - A comparison of the effects of nedocromil sodium and beclomethasone dipropionate on pulmonary function, symptoms, and bronchial responsiveness in patients with asthma. AB - The efficacy of nedocromil sodium (NED) (4 mg twice daily) and beclomethasone dipropionate (BDP) (200 micrograms twice daily) in controlling the symptoms of asthma, the pulmonary function, and bronchial responsiveness to histamine was assessed in a double-blind, double-dummy, crossover study of 39 adult patients with chronic asthma. The patients, most of whom were assessed to be affected to a moderate degree, were insufficiently controlled in their current regimen of inhaled and/or oral bronchodilators. A 2-week baseline period preceded 6 weeks of treatment with each of the study drugs. Both treatment groups demonstrated improvements from baseline in clinical assessment of lung function performed after the first 6 weeks of treatment. No significant differences were observed when the effects of the treatments were compared on FEV1, FVC, and peak expiratory flow. Bronchial reactivity to histamine, measured as the amount of histamine causing a 20% fall in FEV1 (PC20), decreased significantly (p less than 0.05) after 6 weeks of treatment with BDP compared to the effect of NED treatment. Asthma severity, symptom score, and inhaled bronchodilator use demonstrated significantly better results with BDP. After crossover of treatment, the group transferring from NED to BDP continued to improve, whereas the group crossing from BDP to NED tended to demonstrate a major deterioration during the first 3 weeks, after which a stabilization or an increase in FEV1, FVC, and ln PC20 appeared to occur. It is concluded that NED for inhalation is a potent new drug for treatment of both atopic and nonatopic subjects with asthma. With the number of patients and dosages used, the effect on the pulmonary function was not significantly different from that of BDP after the initial 6 weeks of treatment, but BDP had a better effect on asthma severity, overall opinions, symptom score, bronchodilator use, ln PC20, and morning peak expiratory flow. PMID- 2547860 TI - The effects of diet differing in fat, carbohydrate, and fiber on carbohydrate and lipid metabolism in type II diabetes. AB - This study was designed to determine the effects of varying the proportions of carbohydrate, fiber, and fat on metabolic control in Type II diabetes. Ten men, aged 50 to 69 years, with Type II diabetes participated. Four isocaloric diets were consumed for 2 weeks each, with a break of 6 to 14 weeks between diets to ensure no carryover effects. Two of the diets were high in carbohydrate (63% to 65% energy) and low in fat (10% to 12% energy) but differed in their fiber contents (20 vs. 45 gm/day). The other two diets were low in carbohydrate (23% to 27% energy) with either a low or a high fat content (15% vs. 55% energy) and a high or normal protein content (62% vs. 18% energy). The composition of the subjects' usual diets in the week before each of the experimental diets did not vary significantly: carbohydrate 47% to 50% energy, protein 22% to 25% energy, fat 27% to 31% energy, and fiber 24 to 25 gm/day. A 75-gm oral glucose tolerance test and a 12-hour metabolic profile in response to 3 meals typical of the particular diet were conducted before and at the conclusion of each 2-week dietary period. The most significant improvements in metabolic control (as assessed by the effects of the diets on fasting glucose and on lipids, and on the glucose and insulin responses to oral glucose and the mixed meals) were obtained with the high-fiber, high-carbohydrate, low-fat diet and with the low carbohydrate, high-protein, low-fat diet. Metabolic control was not significantly affected by the low-fiber, high-carbohydrate, low-fat diet, but it deteriorated significantly on the low-carbohydrate, high-fat diet. The results of this study confirmed the importance of high fiber and low fat in improving metabolic control in Type II diabetes. In conclusion, if high-carbohydrate, low-fat diets are to be recommended to patients with diabetes, it is essential that the type of carbohydrate recommended be unrefined and high in fiber. PMID- 2547861 TI - In vivo studies on the effect of some insecticides on the hepatic activities of L tryptophan 2,3-dioxygenase and pyridoxal phosphokinase of male mice. AB - The effect of Dimethoate, Carbaryl and Permethrin on the activities of liver L tryptophan 2,3-dioxygenase (total-, holo-, and apo-forms) and pyridoxal phosphokinase of male mice was investigated. Dimethoate inhibited both enzyme systems after single and repeated dose treatment; except the dioxygenase holo enzyme after repeated doses. Single dose of Carbaryl treatment inhibited the pyridoxal phosphokinase, total and holo-enzyme of L-tryptophan dioxygenase. However, the repeated dose treatment do not affect both enzyme systems. Permethrin inhibited only total-, holo- and apo-enzymes of L-tryptophan dioxygenase whereas repeated administration has no significant effect on both enzymes. The data indicate that these insecticides may induce abnormal tryptophan metabolism through which some endogenous bladder carcinogens are formed. PMID- 2547862 TI - Changes of immunological patterns of human cancer cells treated in vitro with a triazene compound. AB - Previous experimental evidence indicates that immunogenicity of mouse tumor cells can be increased by treatment with dacarbazine and other triazene compounds. The present studies have been conducted on the human cell lines H125 (lung cancer), 1246 (melanoma), X3 (EBV-immortalized B cells) subjected to in vitro treatment with 4 (3-methyl-1-triazeno) benzoic acid potassium salt (MTBA). Untreated or drug-treated sublines were tested for susceptibility to allogeneic NK effector cells, either non-stimulated or pretreated with beta-Interferon. In the case of X3 cell line and its MTBA-treated subline the expression of the EBV-associated antigens and the capability of eliciting autologous cytotoxic T lymphocytes (CTL) were also investigated. The results suggest that a modification of membrane antigenic pattern could be induced by MTBA treatment in terms of changes of cell susceptibility to cell-mediated lysis, expression of EBV-related antigens and capability to elicit autologous CTL. PMID- 2547863 TI - Effect of lentinan on superoxide dismutase enzyme activity in vitro. AB - The effects of lentinan on enzyme induced lipid peroxidation, xanthine-xanthine oxidase-induced cytochrome c reduction, and on the superoxide-dismutase (SOD) enzyme activity and expression of human lymphocytes and erythrtocytes were studied. Lentinan in low concentration decreased SOD activity of lymphocytes and erythrocytes from healthy subjects. In higher concentration (10 micrograms/ml) lentinan increased the pathologically low SOD activity of erythrocytes and lymphocytes of patients with cirrhosis of the liver. No significant antioxidant (free radical scavenger) effect has been observed in NADPH-induced and Fe3+ stimulated lipid peroxidation and in xanthine-xanthine oxidase system. PMID- 2547864 TI - Angiotensin II binding to human mononuclear cells. AB - Serum angiotensin converting enzyme activity is elevated in certain human granulomatous diseases. Angiotensin II modestly suppresses thymidine incorporation and augments gamma interferon production from human blood mononuclear cells. This suggests that angiotensin II may play an immunoregulatory role in human granulomatous inflammation. For this reason, the binding of angiotensin II to human peripheral blood mixed mononuclear cells, and to the human monocyte-like cell line, U-937, was studied. Angiotensin II binding to U 937 cells reveals a low level of saturable specific binding (Kd = 3 x 10(-10) M). However, binding to human cells is not easily reversible and is poorly inhibited in a competitive manner. In addition, the molecular integrity of the radioligand is not maintained following binding. Therefore, the definition of classical receptor binding cannot be fulfilled. Since binding is decreased by low temperatures and various metabolic inhibitors, it appears likely that endocytosis occurs, perhaps along with receptor binding. Angiotensin II or its breakdown products modulate the production of monocyte cAMP in spite of the inability to demonstrate classical cell surface receptors. PMID- 2547865 TI - Amebic colitis masquerading as acute inflammatory bowel disease: the role of serology in its diagnosis. AB - Amebic colitis needs to be considered in the differential diagnosis of infectious colitis or inflammatory bowel disease (IBD). Misdiagnosing amebic colitis as idiopathic inflammatory disease may be fatal. Although stool studies remain the initial approach to diagnosis, the incidence of false-negatives associated with this method is high. We report two cases of amebic colitis presenting as inflammatory bowel disease in which the diagnosis of amebic colitis was made by serology. Neither patient had any risk factors for acquiring amebiasis. Paired serology for amebic infections (2-4 weeks apart) should be performed in patients being evaluated for infectious colitis and inflammatory bowel disease. PMID- 2547866 TI - Digoxin-like substance(s) interfere(s) with serum estimations of the drug in cirrhotic patients. AB - We measured the levels of digoxin-like immunoreactivity in the serum of 40 volunteers (20 patients with liver cirrhosis and 20 healthy adults) before and after the administration of a 5-day standard regimen of digoxin. Serum digoxin levels (SDL) were evaluated with two different radioimmunoassay (RIA) kits- Amerlex Digoxin 125I RIA and Digoxin 125I RIA. Digoxin was detectable by each RIA kit in 10 and 15% of controls and 50 and 60% of cirrhotic patients before the administration of the drug, respectively. At the end of the treatment with digoxin, SDL were significantly higher in cirrhotics when compared with those of controls. This study provides evidence that digoxin-like substance(s) is (are) implicated in the detection of high SDL in patients with histologically confirmed liver cirrhosis. PMID- 2547867 TI - Adrenoceptors on blood cells in patients with essential hypertension before and after mental stress. AB - The effect of mental arithmetic (MA) on alpha 2- and beta 2-adrenoceptors on platelets and lymphocytes, respectively, and on plasma catecholamines was studied in normotensive (NT) and essential hypertensive (HT) subjects. There were no significant differences in responses of blood pressure, forearm blood flow (FBF) and heart rate to MA between the two groups. Baseline values and changes in adrenaline and noradrenaline levels during MA were similar in NT and HT. Alpha 2 adrenoceptor density and antagonist affinity did not differ between NT and HT and was not influenced by MA. Baseline values of beta 2-adrenoceptor density also did not differ between NT and HT and increased similarly after MA in both groups. Antagonist affinity to the beta 2-adrenoceptors under baseline conditions was lower in HT and did not change during MA in either group. Our results indicate that there are no differences between NT and HT in alpha 2- and beta 2 adrenoceptor density, either under baseline conditions or after mental stress. PMID- 2547868 TI - The effect of theophylline and cyclic adenosine 3',5'-monophosphate on renin release by afferent arterioles. AB - The effects of theophylline and cyclic adenosine 3',5'-monophosphate (cAMP) on renin release by afferent arterioles were studied. Rabbit afferent arterioles (seven to 10), obtained by a microdissection technique, were incubated for three consecutive 20 min periods in 100 microliters of Medium 199 with 0.1% bovine serum albumin (BSA). Afferent arterioles exposed to theophylline, 1 x 10(-4) mol/l, produced a greater than 100% increase in renin release (0.18 +/- 0.04 to 0.42 +/- 0.05 ng angiotensin l/h per arteriole per h incubation). The renin release stimulated by theophylline was completely abolished by indomethacin and meclofenamate. The stimulation of renin secretion was also blocked when extracellular calcium concentration was decreased to 10(-7) mol/l and the arterioles were permeabilized with calcium ionophore. Dibutyryl cAMP (db-cAMP), 1 x 10(-4) mol/l, and forskolin, 1 mumol/l or 100 mumol/l, failed to stimulate renin release by afferent arterioles. The results of this study therefore suggest that theophylline-stimulated renin release may be mediated through a prostaglandin pathway. The stimulation of renin secretion is also dependent on the extracellular concentration of calcium. PMID- 2547870 TI - Epstein-Barr virus preferentially induces proliferation of primed B cells. AB - EBV can induce human B cells to proliferate, differentiate, and undergo transformation into continuously growing lymphoblastoid cell lines. The EBV responsiveness appears to be confined to a very limited subpopulation of B cells, the nature of which is still unclear. In these studies, we sorted tonsillar B cells on the basis of their expression of the early surface activation Ag, Bac-1, and compared their proliferative responses to EBV. Bac-1+ cells responded to EBV with a relatively high level of DNA synthesis, whereas the Bac-1- cells did not. Both large and small Bac-1+ cells were responsive to EBV and the responsiveness was unrelated to the level of Bac-1 immunofluorescence intensity. Bac-1+ cells were relatively enriched for surface IgM and IgD expression. When the Bac-1- population was enriched for IgM+ cells, the proliferative response was still significantly lower than that of the Bac-1+ population. B cells acquire the ability to bind IgM relatively late after activation, and this feature did not distinguish the EBV-responsive B cells. The results suggest B cells become responsive to EBV after an early activation signal. PMID- 2547869 TI - IL-4 induces LFA-1 and LFA-3 expression on Burkitt's lymphoma cell lines. Requirement of additional activation by phorbol myristate acetate for induction of homotypic cell adhesions. AB - LFA-1 and LFA-3 expression is absent or low on Burkitt's lymphoma cell lines and low on the EBV-transformed B cell line UD61. Incubation of cells of BL2 and of UD61 with various concentrations of IL-4 resulted in induction of LFA-1 and LFA-3 expression in a dose dependent fashion. This effect was already observed after 16 h of incubation whereas maximal expression was obtained after 72 h. Induction of LFA-1 and LFA-3 expression seemed to be specific for IL-4, because IL-1, IL-2, IL 3, IFN-alpha, IFN-gamma and a low m.w. B cell growth factor were ineffective. LFA 1 and LFA-3 induction by IL-4 was blocked specifically by an anti-IL-4 antiserum. Induction of LFA-1 expression by IL-4 was furthermore confirmed at the specific LFA-1 beta-chain mRNA level. IL-4 was unable to induce LFA-1 expression on EBV transformed lymphoblastoid cell lines of two LFA-1-deficient patients. BL2 grows as single cells, but induction of LFA-1 and LFA-3 expression by IL-4 was insufficient to induce homotypic cell adhesions and required PMA as a second signal. PMA alone did not induce LFA-1 antigen expression and was unable to induce adhesions between BL2 cells in the absence of IL-4 in 22 h assays. Addition of PMA to BL2 cells that expressed LFA-1 Ag upon incubation with IL-4 resulted in aggregate formation within 30 min. Adhesions between BL2 cells induced by IL-4 in combination with PMA were blocked by anti-LFA-1 beta or anti LFA-1 alpha-chains mAb. In addition, these mAbs dispersed preformed aggregates of BL2 cells. Our results indicate that IL-4 can induce the adhesion molecules LFA-1 and LFA-3 on B cell lines, but that an additional activation signal provided by PMA was required for the induction of homotypic cell adhesions. PMID- 2547871 TI - Effects of tumor necrosis factor and epidermal growth factor on cell morphology, cell surface receptors, and the production of tissue inhibitor of metalloproteinases and IL-6 in human microvascular endothelial cells. AB - The effect of human TNF on cultured human microvascular endothelial (HME) cells was examined. Incubation with TNF alone transformed the morphology of HME cells from a cobblestone-like appearance into a disordered array of criss-crossed, elongated, spindle-shaped cells. Coadministration of epidermal growth factor (EGF) and TNF caused even more dramatic morphologic changes than TNF alone. Addition of basic fibroblast growth factor or insulin-like growth factor-I showed rather weak effects on cell morphology than EGF. Cell growth of HME cells was stimulated up to two-fold by TNF whereas addition of EGF additively enhanced the growth rate. Treatment of HME cells with 10 ng/ml EGF increased the binding of 125I-TNF, and Scatchard analysis showed increased TNF-R number by EGF treatment. Cellular response to TNF in the absence or presence of EGF was assessed by analyzing SDS-PAGE patterns of secreted proteins from HME cells. TNF enhanced the secretion of a protein of molecular weight 25,000 Da (25 kDa) which was found to be IL-6. In contrast, secretion of a polypeptide of 29 kDa was significantly increased when HME cells were treated with EGF, but not with TNF. Coadministration of TNF and EGF synergistically increased the secretion of the 29 kDa protein. This 29-kDa protein was found to be tissue inhibitor of metalloproteinases when assayed with antitissue inhibitor of metalloproteinases antibody. TNF and EGF also enhanced secretion of collagenase with Mr of approximately 55 kDa. Increased steady state levels of the inhibitor mRNA were observed when HME cells were treated with EGF, and coadministration of TNF further increased the levels. The morphologic transformation of HME cells by TNF and/or EGF is discussed in relation to their expression of the secreted proteins. PMID- 2547872 TI - Involvement of guanine nucleotides in superoxide release by fluoride-treated neutrophils. Implications for a role of a guanine nucleotide regulatory protein. AB - Previous studies demonstrating hydrolysis of phosphatidylinositol bisphosphate (PIP2) and generation of inositol phosphates in neutrophils exposed to 20.0 mM NaF provide indirect evidence that activation of phospholipase-associated guanine nucleotide regulatory protein, a guanine nucleotide binding protein which regulates the activation of a membrane inositol-specific phospholipase C, is an early event in the neutrophil stimulus-response pathway triggered by fluoride. Consistent with this hypothesis, exposure of a plasma membrane rich preparation isolated from 32P labeled neutrophils to 20.0 mM NaF resulted in hydrolysis of labeled PIP2. Levels of other phospholipids were not affected. Inositol bisphosphate and inositol trisphosphate were detected in extracts of neutrophil plasma membranes exposed to fluoride. To further explore the involvement of guanine nucleotides in functional responses of intact neutrophils triggered by fluoride, we preincubated cells with 2-beta-D-ribofuranosylthiazole-4-carboxamide (tiazofurin), a selective inhibitor of inosine monophosphate dehydrogenase, to diminish guanine nucleotide synthesis and then compared superoxide generation induced by FMLP, PMA, digitonin, and 20.0 mM NaF to intracellular levels of guanine nucleotides. Preincubation of neutrophils for 2.5 h at 37 degrees C with tiazofurin resulted in dose-dependent depletion of GTP and GDP. Maximal depletion of guanine nucleotides required relatively high levels of tiazofurin (200 to 400 microM) and resulted in a 55 to 60% reduction of GTP and GDP. The effects of tiazofurin on guanine nucleotides levels were not observed when neutrophils were preincubated at 4 degrees C. AT 37 degrees C, tiazofurin also decreased intracellular ATP and ADP levels but adenine nucleotide depletion was less pronounced than guanine nucleotide depletion for each concentration of tiazofurin used. When tiazofurin was removed by washing cells after incubation, adenine nucleotide quickly returned to preincubation values but guanine nucleotide levels remained depressed. Addition of exogenous guanosine (200 microM) prevented tiazofurin-dependent depletion of guanine nucleotides but had no influence on adenine nucleotide depletion. Superoxide released triggered by FMLP and F- was inhibited to an extent similar to that of guanine nucleotide depletion under different conditions of preincubation. Inhibition of superoxide release was not observed if cells were preincubated at 4 degrees C, was not rapidly reversible, and was not observed when guanosine was added with tiazofurin.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2547873 TI - Protease-modulation of neutrophil superoxide response. AB - Although prior studies with mAb have defined an endogenous chymotrypsin-like protease in the neutrophil (polymorphonuclear leukocyte (PMN)) membrane that is associated with initiation of superoxide response to inflammatory stimuli, it is not known whether extracellular proteases (in the inflammatory milieu) can also influence PMN activation. This study examined the ability of four neutral proteases: cathepsin G, elastase, chymotrypsin, and trypsin, to modify PMN superoxide response to FMLP, PMA, and arachidonate. In response to 1 microM FMLP, PMN treated with cathepsin G, chymotrypsin, or elastase showed 64%, 60%, and 32% increases, respectively, in superoxide generation when compared with control, untreated cells (p less than 0.05 for each). These increments were dependent on intact enzymatic function of the proteases, were greatest when enzyme and stimulus were added concurrently, and persisted after PMN were washed free of enzyme. Enhancement of superoxide response was not stimulus specific; in response to 10 ng/ml PMA, cells treated with cathepsin G showed a 84%, and elastase a 57%, increase in superoxide generation (p less than 0.05 for both) with a marked reduction in the time required for onset of this response. For cell activation with 80 microM arachidonate, treatment with elastase produced a 180% increase in superoxide production (p less than 0.025). Neutrophils incubated with trypsin demonstrated significant decreases in superoxide response to PMA (-34%, p less than 0.05) and arachidonate (-39%, p less than 0.01). The enzymes themselves were not stimuli for superoxide production nor were they scavengers for superoxide in cellfree system. We conclude that local release of the PMN primary-granule neutral proteases, cathepsin G, and elastase within inflammatory sites can augment neutrophil effector function by up-regulating oxidative response to defined inflammatory stimuli. This autocrine/paracrine function may provide a significant increase in antimicrobial activity, but may also enhance the potential for host tissue injury. PMID- 2547874 TI - Measurement of binding specificity between T cell lymphomas and murine leukemia viruses. AB - We have previously reported the presence of receptors on radiation leukemia virus (RadLV)-induced thymomas and malignant thymocytes from AKR mice which specifically bind retrovirus produced by these T cell clones. These receptors have been shown to have specificity for virus reminiscent of an immune-specific receptor. Previous studies on T cell lymphoma binding to retroviruses have involved measurement of the interaction of labelled virus with cells using fluorescence-activated cell sorter (FACS) analysis (McGrath et al., J. Virol. (1978) 25, 923; McGrath and Weissman, Cell (1979) 17, 65; Weissman and McGrath, Curr. Top. Microbiol. Immunol. (1982) 98, 103). Here we report development of an assay for measuring lymphoma binding to virus, prepared as an immunoabsorbent adhered to a microtiter plate. Using this assay, we have shown that only T and not B cell lymphomas can bind to T cell-tropic viruses, and some cell lines have greatest specificity for homologous virus. The AKR-derived T cell lymphomas, SL3 and KKT-2, show greater specificity for leukemogenic AKR viruses, than for an AKR xenotropic virus or the recombinant AKR virus, MCF247. The RadLV-induced T cell lines, C6VL/1 and BL/VL3, have been found to bind cross-reactively to several different murine leukemia viruses (MuLVs). RadLV-induced T cell lymphomas do have greater specificity for their cognate retroviruses since free, homologous retrovirus can best block the interaction between cells and virus adhered to the wells of a microtiter plate. Cross-reactive interactions are more easily demonstrated by this assay, probably because low avidity interactions are stabilized as a result of the mode of virus presentation. Binding specificity for retroviral envelope determinants has been demonstrated using a rat anti retroviral antiserum prepared as an F(ab)1 fragment. This antiserum can inhibit the interaction between the C6VL/1 thymoma and its RadLV virus. Specificity of this antibody for a gp70-like protein was confirmed by NaDodSO4-polyacrylamide gel electrophoresis (PAGE) and by loss of this activity after absorption of antibody on virus. Antibodies specific for RadLV/VL3 gp70 determinants can inhibit the interaction of C6VL/1 with RadLV/VL3 suggesting that cross-reactive binding to heterologous virus is also specific for a gp70 viral env determinant. PMID- 2547875 TI - Purification of human blood eosinophils by negative selection using immunomagnetic beads. AB - A simple method for isolating highly purified eosinophils from human blood is described. Buffy coats from normal individuals (eosinophil counts less than 0.4 x 10(9)/litre) were centrifuged through a two layer Percoll density gradient, to produce a granulocyte fraction containing neutrophils and eosinophils. Neutrophils were extracted from this fraction using a monoclonal antibody (CLB FcR gran 1) against CD 16 (Fc gamma R III) in a direct or indirect selection procedure using immunomagnetic beads (Dynabeads). This negative immunoselection produced eosinophils of greater purity and with a superior capacity to mount a respiratory burst than eosinophils isolated by a method employing metrizamide. PMID- 2547876 TI - The ethics of using human volunteers for high-risk research. PMID- 2547877 TI - Reactivation of Toxoplasma gondii by cytomegalovirus disease in mice: antimicrobial activities of macrophages. AB - Reactivation of Toxoplasma gondii infection often occurs concurrently with active cytomegalovirus (CMV) disease in immunocompromised patients, and CMV disease results in immunosuppression. To determine if murine CMV (MCMV) infection decreases resistance to T. gondii, mice with latent T. gondii infection were infected with MCMV. T. gondii infection reactivated, manifested primarily as pneumonia. Lung macrophages supported the growth of T. gondii before, during, and after T. gondii reactivation. Peritoneal macrophages inhibited the growth of T. gondii as pneumonia developed and became permissive as pneumonia resolved. Mice with latent T. gondii infection could survive larger doses of MCMV than could controls. Thus, MCMV infection led to reactivation of latent T. gondii infection in mice. Activation of lung macrophages, assessed by their ability to inhibit replication of T. gondii in vitro, was not associated with control of T. gondii infection. PMID- 2547878 TI - Respiratory failure complicated by adenovirus serotype 29 in a patient with AIDS. PMID- 2547879 TI - Prevalence of antibody to human T cell leukemia virus type 1 (HTLV-1) in populations of Ivory Coast, West Africa. AB - A large cross-sectional serologic survey for human T cell leukemia virus type 1 (HTLV-1) antibody was conducted in 3,177 Ivory Coast residents to evaluate the prevalence of HTLV-1 and to determine possible risk factors and correlates of HTLV-1 infection. Of the 3,177 serum samples, 110 (3.5%) were positive for antibody to HTLV-1 by indirect immunofluorescence assay and Western blot. The prevalence of HTLV-1 antibody in the general adult population was 1.8% and increased significantly with age. No difference between males (1.5%) and females (2%) was found. The highest prevalences were observed in female prostitutes (7.4%), patients with neurologic syndromes (5.8%), and lepers (13.7%). The high prevalence of HTLV-1 infection in prostitutes suggests that heterosexual contact is involved in the transmission of HTLV-1 and that prostitutes could play an important role in the spread of the virus in Africa. The high prevalence of HTLV 1 in patients with neurologic syndromes confirms the association between HTLV-1 and some type of neuropathies, as has been observed in the West Indies and Japan. The high prevalence observed in lepers deserves further investigation to find the cause of the association. Twenty-five individuals, including prostitutes, were coinfected with HTLV-1 and human immunodeficiency virus (HIV). Prospective studies are necessary to evaluate the exact role of HTLV-1 alone or in combination with HIV in inducing specific diseases. PMID- 2547880 TI - Group C rotavirus infections in patients with diarrhea in Thailand, Nepal, and England. AB - Atypical rotavirus obtained from fecal specimens of six patients with diarrhea from Thailand, Nepal, and England were characterized by using polyacrylamide gel electrophoresis and immune electron microscopy. The electropherotypes were characteristic of the porcine reference group C rotavirus strain but demonstrated considerable strain-to-strain variation. Human convalescent group C sera had a high titer (1:320) when tested against the human isolates and a low titer (1:40) when tested against a porcine reference strain (Cowden). When porcine antiserum (Cowden) was tested against the human isolates, the titers ranged from 1:40 to 1:320, indicating significant antigenic diversity between strains. Group C rotavirus appears to have a worldwide distribution as an agent associated with diarrhea in children and adults. PMID- 2547881 TI - Interactions between simian immunodeficiency virus and Mycobacterium leprae in experimentally inoculated rhesus monkeys. AB - Thirty-four rhesus monkeys were inoculated with Mycobacterium leprae inoculum isolated from sooty mangabey monkeys with leprosy. Later it was learned that one of the M. leprae-donor mangabeys was asymptomatically infected with simian immunodeficiency virus (SIV). Thus, five of the rhesus monkey were coinoculated with M. leprae and SIV. Three of the five became SIV-positive and developed signs of leprosy and an AIDS-like illness. Two animals remained healthy. The coinoculated leprosy-positive rhesus monkeys developed leprosy despite serologic response patterns to M. leprae antigens that usually indicate leprosy resistance. Three (60%) of the five SIV-positive rhesus monkeys developed leprosy compared with 21% of the animals who received SIV-free M. leprae inocula. Diminished lepromin skin test responses and decreasing T-helper cell percentages were observed in SIV-coinoculated rhesus monkeys with leprosy. These observations suggest that SIV increases the susceptibility of rhesus monkeys to leprosy, possibly related to loss of T-helper cell function. PMID- 2547882 TI - Herpes zoster in an adult recipient of live attenuated varicella vaccine. AB - A healthy 30-y-old female physician who was immunized with two doses of live attenuated varicella vaccine developed a localized case of zoster involving the right T8-10 dermatomes 36 mo after vaccination. The virus isolated from her rash was an unusual wild-type of varicella-zoster virus. After immunization she developed detectable antibodies to varicella-zoster virus, but antibodies were no longer detectable after 20 mo. Six months before development of zoster, she was exposed to a patient with varicella; however, she did not develop a clinical illness although she again became seropositive. To date, this is the only reported case of zoster among 187 healthy adult vaccinees. PMID- 2547883 TI - Virus induced immune interferon contains both interferon-alpha and -gamma. PMID- 2547884 TI - Chronic active Epstein-Barr virus (EBV) infection is associated with clonotypic intracellular terminal regions of the EBV. PMID- 2547885 TI - Prevalence of antibodies to human herpesvirus 6 in human immunodeficiency virus 1 seropositive and -negative intravenous drug addicts. PMID- 2547886 TI - Polymorphism of human herpesvirus 6 DNA from five Japanese patients with exanthem subitum. PMID- 2547887 TI - Herpes simplex type 2 pericarditis and bilateral facial palsy in a patient with AIDS. PMID- 2547888 TI - Adenoid cystic carcinoma of the salivary glands: clinicopathological survey of 51 patients. AB - A clinicopathological study of 51 cases of adenoid cystic carcinoma in the head and neck region seen over a period of 20 years is presented. The correlation between various histological features and their impact on the results is attempted. Longterm results with respect to loco-regional recurrences, distant metastases and survival with various modalities of therapy are presented. Although this is a small series, radical surgery in keeping with the aim of avoiding undue mutilation, thus preserving the quality of life, followed by a full course of radiotherapy has been the mainstay of treatment and the results here presented justify this approach. PMID- 2547889 TI - Human neutrophils activated by interferon-gamma and tumour necrosis factor-alpha kill Entamoeba histolytica trophozoites in vitro. AB - The interaction between cytokine-activated human neutrophils and Entamoeba histolytica trophozoites was studied as well as the mechanism(s) involved. Treatment of neutrophils with rIFN-gamma alone allowed them to kill 30% of E. histolytica trophozoites; however, rIFN-gamma and rTNF-alpha pretreatments in combination increased neutrophil killing to 70%. In the absence of direct contact between neutrophils and amebae, rIFN-gamma-treated neutrophils were shown to kill 70% of amebae, and rIFN-gamma- and rTNF-alpha-treated neutrophils killed 97% of amebae. Neutrophil enhancement of amebicidal activity following cytokine treatments was correlated with increased neutrophil resistance to amebic contact dependent killing and was shown to be 73% H2O2 dependent. PMID- 2547890 TI - Detection of vesicular lipoproteins in lecithin:cholesterol acyltransferase deficient plasma by 1H-NMR spectroscopy. AB - The proton NMR spectra of the N-methyl choline region of normal and lecithin:cholesterol acyltransferase (LCAT)-deficient lipoproteins and of egg yolk phosphatidylcholine-cholesterol 55:45 (mol %) vesicle mixtures have been examined in the presence and absence of manganous sulfate as a line-broadening reagent. Manganous ions quenched all of the signal arising from normal lipoproteins and only part of the vesicle signal corresponding to the outer monolayer. There was no net loss of vesicular phospholipid when vesicles were added to normal lipoproteins and as little as 5% (or 100 micrograms) of the vesicular phospholipid could be detected and quantitated in the mixture of lipoproteins. Similar experiments performed on plasma lipoproteins from an LCAT deficient patient indicated that 42% of the phospholipid was associated with vesicular lipoproteins. These experiments demonstrate that this technique can be used to detect and quantify small amounts of vesicular structures directly in a mixture of micellar lipoproteins. PMID- 2547891 TI - Studies on the stability and detection of cocaine, benzoylecgonine, and 11-nor delta-9-tetrahydrocannabinol-9-carboxylic acid in whole blood using Abuscreen radioimmunoassay. AB - A study was undertaken to assess the stability and the radioimmunoassay (RIA) detection of cocaine, benzoylecgonine (BZE), and 11-nor-delta-9 tetrahydrocannabinol-9-carboxylic acid (THC-COOH) in whole blood while stored in 4 different kinds of blood collection tubes for up to 30 days at refrigeration and room temperatures. At various intervals, the tubes were sampled and analyzed using Abuscreen RIA. Also, semi-quantitative data derived from RIA analysis of forensic blood specimens were compared with quantitative data acquired using gas chromatography (GC) or GC/mass spectrometry (GC/MS) on the same specimens. RIA and chromatographic studies revealed that BZE and THC-COOH were stable in blood under all conditions studied. Cocaine, however, was found not to be stable in blood, especially when stored at room temperatures. Despite cocaine's instability in blood, RIA was able to detect the presence of cocaine and its breakdown products in blood under all conditions studied. PMID- 2547892 TI - [3H]MK-801 binding sites in postmortem brain regions of schizophrenic patients. AB - [3H]MK-801 binding was used as a marker for the NMDA receptorion channel complex in postmortem brain samples from the frontal cortex, hippocampus, putamen, entorhinal region, and amygdala of schizophrenic patients and controls. In schizophrenia [3H]MK-801 binding levels were increased in all brain regions investigated reaching significance in the putamen. PMID- 2547893 TI - Syntheses and spectroscopic studies of potential antitumor copper(II) complexes with 5-phenylazo-3-methoxy salicylidene thiosemicarbazone and N4 substituted thiosemicarbazones. AB - Five new copper(II) complexes of 5-phenylazo-3-methoxy salicylidene thiosemicarbazone and N4 substituted thiosemicarbazones have been synthesized. They have been characterized by chemical analyses, magnetic, conductance data, and by ultraviolet (UV)--visible, infrared, and electron spin resonance spectra. The complexes have the general formula CuL2, where HL is the ligand. One representative complex has been screened in vitro and in vivo against P388 lymphocytic leukemia cells sensitive and resistant to adriamycin (P388/S and P388/R). It has shown promising growth inhibition activity. We are reporting here for the first time the antineoplastic activity of this complex against experimental tumor systems. PMID- 2547894 TI - Mechanistic aspects of the dithiocarbamate-induced mobilization of cadmium. AB - The examination of some of the species involved in the in vivo processes in which dithiocarbamates mobilize cadmium from its intracellular deposits indicates that several competing reactions occur. Rates of hydrolytic decomposition of a series of dithiocarbamates capable of mobilizing cadmium in vivo have been determined, and the solubility behavior and nuclear magnetic resonance (NMR) spectra of their cadmium complexes have been examined. Some of the dithiocarbamates most effective in this mobilization process are shown to undergo slow conversion to oxazolidine 2-thiones in the presence of cadmium. All of the cadmium complexes involved in the mobilization process are shown to undergo rapid ligand exchange. While dissociative mechanisms based on the turnover of metallothionein are inconsistent with the experimental data, at least two associative mechanisms are possible. These involve attack on the metallothionein by the dithiocarbamate itself or by a compound derived from it by known metabolic processes. PMID- 2547895 TI - Synthesis and reactivity of the oxovanadium(IV) complexes of two N-O donors and potentiation of the antituberculosis activity of one of them on chelation to metal ions: Part IV. AB - A new series of oxovanadium(IV) complexes of two aromatic acidhydrazides (BH and AH) have been reported. Of these two donors, AH is known to possess considerable in vitro antitubercular activity. At pH 2-4, oxometal complexes of the type [VO(BH/AH)2SO4].nH2O (n = 1, 0) and [VO(BH/AH)(C2O4)H2O].H2O (BH = C6H5CONHNH2 and AH = (2-NH2)C6H4.CO.NHNH2) were obtained. Reactions of [VO(BH/AH)(C2O4)H2O].H2O with a monodentate Lewis base lead to the isolation of metal-ligand complexes [VO(BH/AH)(C2O4)L].nH2O (L = NH3, n = 1, L = py, n = 2). Disposition of the bonding sites of donor molecules around the oxometal acceptor center and status of the metal-oxygen multiple bond have been established. A monomeric and distorted octahedral donor environment for the oxovanadium(IV) ion has been proposed on the basis of the electron paramagnetic resonance (EPR) spectra and magnetic susceptibility measurements. Antitubercular activities, in vitro, of the oxovanadium(IV) complexes of AH have also been evaluated towards tuberculosis mycobacteria such as Mycobacterium flae, Mycobacterium smegmatis and Mycobacterium H37Rv. PMID- 2547896 TI - Triamminemonoaquocobalt(III) complexes of pyrophosphate and adenosine diphosphate (ADP). AB - Cobalt(III)H2O(NH3)3 pyrophosphate has been shown by proton and 31P nuclear magnetic resonance (NMR) to be a facial bidentate complex. Cobalt(III)H2O(NH3)3 adenosine diphosphate has been resolved into lambda and delta isomers by chromatography on cycloheptaamylose. Both the lambda and delta forms are a pair of isomers that are not separated by cycloheptaamylose, reverse phase high pressure liquid chromatography (HPLC), or cation exchange chromatography. These isomers presumably represent syn- and anti-arrangement of coordinated water and adenosine. PMID- 2547897 TI - Evidence for the presence of serotonin receptors negatively coupled to adenylate cyclase in the rabbit iris-ciliary body. AB - Serotonin has no obvious effect on basal cyclic AMP levels but reduces the forskolin-, isoproterenol-, and vasoactive intestinal peptide-induced stimulation of cyclic AMP levels in a dose-dependent manner. Serotonergic, cholinergic, muscarinic, alpha-adrenergic, and dopaminergic antagonists have no effect on the serotonin response. Topical application of a serotonin/pargyline solution to the living eye causes desensitisation of the serotonin response in the iris-ciliary body, an observation confirming the presence of specific serotonergic receptors linked to adenylate cyclase. The 5-HT1A [5-hydroxytryptamine (serotonin) type 1A] receptor agonists 8-hydroxy-2-(di-n-propylamino)tetralin and buspirone mimic the serotonin response in reducing the forskolin-stimulated cyclic AMP levels, as do the indole derivatives 5-methoxytryptamine, 5-hydroxtryptophan, and tryptamine. However, the ineffectiveness of the 5-HT1A agonist ipsapirone and the inability of spiroxatrine to block the serotonin response show that classical 5-HT1A receptors are not involved. The serotonin response is blocked by pertussis toxin and is insensitive to the phosphodiesterase inhibitor theophylline, which indicates the involvement of an inhibitory guanine regulatory protein in the coupling of the serotonin receptor to the adenylate cyclase catalytic unit. PMID- 2547898 TI - Beta-N-oxalylamino-L-alanine action on glutamate receptors. AB - beta-N-Oxalylamino-L-alanine (L-BOAA) is a non-protein excitatory amino acid present in the seed of Lathyrus sativus L. This excitotoxin has been characterized as the causative agent of human neurolathyrism, an upper motor neuron disease producing corticospinal dysfunction from excessive consumption of the lathyrus pea. Previous behavioral, tissue-culture, and in vitro receptor binding investigations revealed that L-BOAA might mediate acute neurotoxicity through quisqualate (QA)-preferring glutamate receptors. The present study demonstrates the stereospecific action of L-BOAA on glutamate receptor binding in whole mouse brain synaptic membranes. L-BOAA was most active in displacing thiocyanate (KSCN)-sensitive specific tritiated (RS)-alpha-3-hydroxy-5 methylisoxazole-4-propionic acid (AMPA) binding (i.e., QA receptor) (Ki = 0.76 microM) with a rank-order potency of QA greater than kainate greater than N methyl-D-aspartate (NMDA). By contrast, the nonneurotoxic D-BOAA isomer (100 microM) was essentially inactive in displacing radioligands for glutamate receptors, except the NMDA site, where it was equipotent with L-BOAA. Scatchard analysis of L-BOAA displacement of specific [3H]AMPA binding indicated competitive antagonism (KD: control, 135 nM; L-BOAA, 265 nM) without a significant change in QA-receptor density, and Hill plots yielded coefficients approaching unity. Differential L-BOAA concentration-dependent decreases in specific [3H]AMPA binding were observed in synaptic membranes, indicating that the neurotoxin was more potent in displacing specific binding from frontal cortex membranes, followed by that for corpus striatum, hippocampus, cerebellum, and spinal cord. (ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547899 TI - Thiamine triphosphatase in the membranes of the main electric organ of Electrophorus electricus: substrate-enzyme interactions. AB - The main electric organ of Electrophorus electricus is particularly rich in thiamine triphosphate (TTP). Membrane fractions prepared from this tissue contain a thiamine triphosphatase that is strongly activated by anions and irreversibly inhibited by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), an anion transport inhibitor. Kinetic parameters of the enzyme are markedly affected by the conditions of enzyme preparation: In crude membranes, the apparent Km is 1.8 mM and the pH optimum is 6.8, but trypsin treatment of these membranes or their purification on a sucrose gradient decreases both the apparent Km (to 0.2 mM) and the pH optimum (to 5.0). Anions such as NO3- (250 mM) have the opposite effect, i.e., even in purified membranes, the pH optimum is now 7.8 and the Km is 1.1 mM; at pH 7.8, NO3- increases the Vmax 24-fold. TTP protects against inhibition by DIDS, and the KD for TTP could be estimated to be 0.25 mM, a value close to the apparent Km measured in the same purified membrane preparation. Thiamine pyrophosphate (0.1 mM) did not protect against DIDS inhibition. At lower (10(-5) 10(-6) M) substrate concentrations, Lineweaver-Burk plots of thiamine triphosphatase activity markedly deviate from linearity, with the curve being concave downward. This suggests either anticooperative binding or the existence of binding sites with different affinities for TTP. The latter possibility is supported by binding data obtained using [gamma-32P]TTP. Our data suggest the existence of a high-affinity binding site (KD of approximately 0.5 microM) for the Mg-TTP complex.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547900 TI - Islet-activating protein inhibits the beta-adrenergic receptor facilitation elicited by gamma-aminobutyric acidB receptors. AB - gamma-Aminobutyric acidB (GABAB) receptor recognition sites that inhibit cyclic AMP formation, open potassium channels, and close calcium channels are coupled to these effector systems by guanine nucleotide binding proteins (G proteins). These G proteins are ADP-ribosylated by islet-activating protein (IAP), also known as pertussis toxin. This process prevents receptor coupling to these G proteins. In slices of cerebral cortex and hippocampus from rat, stimulation of GABAB receptors with baclofen, a receptor agonist, also potentiates the accumulation of cyclic AMP stimulated by beta-adrenergic agonists. It was unknown whether those GABAB receptors that potentiate the beta-adrenergic response were also sensitive to IAP. IAP was injected intracerebroventricularly into rats to ADP-ribosylate IAP-sensitive G proteins. Four days after the IAP injection, 38% and 52% of these G proteins from cerebral cortex and hippocampus, respectively, were ADP ribosylated by the IAP injection. In slices of both structures prepared from IAP treated rats, the GABAB receptor-mediated potentiation of the beta-adrenergic receptor response was attenuated. Thus, many GABAB receptor-mediated responses are coupled to IAP-sensitive G proteins. PMID- 2547901 TI - Selective inhibition of nerve growth factor-stimulated protein kinases by K-252a and 5'-S-methyladenosine in PC12 cells. AB - K-252a, a protein kinase inhibitor isolated from the culture broth of Nocardiopsis sp., inhibits the nerve growth factor (NGF)-stimulated phosphorylation of microtubule-associated protein 2 (MAP2) and Kemptide (synthetic Leu-Arg-Arg-Ala-Ser-Leu-Gly) by blocking the activation of two independent kinases in PC12 cells: MAP2/pp250 kinase and Kemptide kinase. The NGF stimulated activation of these kinases is inhibited in a dose-dependent manner following treatment of the cells with K-252a. Although these kinases also are activated by epidermal growth factor (EGF) and 12-O-tetradecanoyl-phorbol 13 acetate, K-252a has no inhibitory effect when these agents are used. Half-maximal inhibition of the activation of both kinases was observed at 10-30 nM K-252a. K 252a was shown to directly inhibit the activity of MAP2/pp250 kinase and Kemptide kinase when added to the phosphorylation reaction mixture in vitro; however, half maximal inhibition under these conditions was observed at greater than or equal to 50 nM K-252a. These data suggest that K-252a exerts its effects at a step early in the cascade of events following NGF binding. The effects of K-252a are similar to those reported for 5'-S-methyladenosine (MTA) and other methyltransferase inhibitors. Treatment of PC12 cells with MTA inhibited NGF-, but not EGF-mediated activation of MAP2/pp250-kinase (Ki greater than 500 microM). MTA, when added to the phosphorylation reaction mixture in vitro, directly inhibited kinase activity (Ki = 50 microM), suggesting that the effects of MTA may be the result of its action on protein kinases rather than methyltransferases. PMID- 2547902 TI - Regulatory properties of calcium/calmodulin-dependent protein kinase II in rat brain postsynaptic densities. AB - Calcium/calmodulin (CaM)-dependent protein kinase II (CaM-kinase II) contained within the postsynaptic density (PSD) was shown to become partially Ca2+ independent following initial activation by Ca2+/CaM. Generation of this Ca2+ independent species was dependent upon autophosphorylation of both subunits of the enzyme in the presence of Mg2+/ATP/Ca2+/CaM and attained a maximal value of 74 +/- 5% of the total activity within 1-2 min. Subsequent to the generation of this partially Ca2+-independent form of PSD CaM-kinase II, addition of EGTA to the autophosphorylation reaction resulted in further stimulation of 32PO4 incorporation into both kinase subunits and a loss of stimulation of the kinase by Ca2+/CaM. Examination of the sites of Ca2+-dependent autophosphorylation by phosphoamino acid analysis and peptide mapping of both kinase subunits suggested that phosphorylation of Thr286/287 of the alpha- and beta-subunits, respectively, may be responsible for the transition of PSD CaM-kinase II to the Ca2+ independent species. A synthetic peptide 281-309 corresponding to a portion of the regulatory domain (residues 281-314) of the soluble kinase inhibited syntide 2 phosphorylation by the Ca2+-independent form of PSD CaM-kinase II (IC50 = 3.6 +/- 0.8 microM). Binding of Ca2+/CaM to peptide 281-309 abolished its inhibitory property. Phosphorylation of Thr286 in peptide 281-309 also decreased its inhibitory potency. These data suggest that CaM-kinase II in the PSD possesses regulatory properties and mechanisms of activation similar to the cytosolic form of CaM-kinase II. PMID- 2547903 TI - Muscarinic and quisqualate receptor-induced phosphoinositide hydrolysis in primary cultures of striatal and hippocampal neurons. Evidence for differential mechanisms of activation. AB - Several neurotransmitters activate polyphosphoinositide (PPI) hydrolysis in CNS neurons as the first step of a transmembrane signalling cascade that may lead to neuronal circuit modulation. Muscarinic and quisqualate receptor-triggered PPI hydrolysis was investigated in neuronal primary cultures. A clear increase in inositol phosphates (Ins-Ps) was detected as early as 15 s after the agonist addition; at this time, the increases of inositol 1,4,5-trisphosphate (measured by HPLC) were relatively larger with respect to the other Ins-Ps. Ins-P accumulation was maintained in part in a Ca2+-free medium, excluding that Ca2+ entry is the fundamental step of the receptor-induced PPI hydrolysis. Acute cell pretreatment with phorbol dibutyrate, an activator of protein kinase C, was able to inhibit 50% of the response to carbachol, and almost completely the quisqualate effect, suggesting a negative feedback modulation by the enzyme. Finally, pertussis toxin failed to inhibit muscarinic responses, whereas it blocked greater than 70% of the quisqualate stimulation. The two receptors therefore appear coupled to phosphodiesterase by two different G proteins. The comparison of the results obtained by stimulating the two receptor systems suggests that the generation of the same intracellular signal at two distinct receptor types may occur by different coupling mechanisms, and be differently regulated even in the same neuronal preparations. PMID- 2547904 TI - Antiproliferative action of benzodiazepines in cultured brain cells is not mediated through the peripheral-type benzodiazepine acceptor. AB - The benzodiazepines, Ro 5-4864, diazepam, clonazepam, and also PK-11195, inhibited, at micromolar concentrations, the proliferation of rat C6 glioma and mouse neuro-2A neuroblastoma cells in culture. The cells possessed high levels of "peripheral-type" high-affinity benzodiazepine binding sites as judged by binding assays and displacement potencies. However, the different potencies and specificities of compounds for the antiproliferative actions and binding affinities for the binding site suggest that the antiproliferative actions were not mediated through the peripheral-type binding site. In support of this, these compounds have also been shown to inhibit proliferation of some nonneuronal cultured cell lines, e.g., mouse SP2/O-Ag 14 hybridoma and rat NCTC epithelial cells, which have no detectable high-affinity peripheral-type benzodiazepine binding sites. PMID- 2547905 TI - Light-induced dopamine release from teleost retinas acts as a light-adaptive signal to the retinal pigment epithelium. AB - In the retinal pigment epithelium (RPE) of lower vertebrates, melanin pigment granules migrate in and out of the cells' long apical projections in response to changes in light condition. When the RPE is in its normal association with the retina, light onset induces pigment granules to disperse into the apical projections; dark onset induces pigment granules to aggregate into the cell bodies. However, when the RPE is separated from the retina, pigment granule movement in the isolated RPE is insensitive to light onset. It thus seems likely that a signal from the retina communicates light onset to the RPE to initiate pigment dispersion. We have examined the nature of this retina-to-RPE signal in green sunfish, Lepomis cyanellus. In isolated retinas with adherent RPE, light induced pigment dispersion in the RPE is blocked by treatments known to block Ca2+-dependent transmitter release in the retina. In addition, the medium obtained from incubating previously dark-adapted retinas in the light induces light-adaptive pigment dispersion when added to isolated RPE. In contrast, the medium obtained from incubating dark-adapted retinas in constant darkness does not affect pigment distribution when added to isolated RPE. These results are consistent with the idea that RPE pigment dispersion is triggered by a substance that diffuses from the retina at light onset. The capacity of the conditioned medium from light-incubated retinas to induce pigment dispersion in isolated RPE is inhibited by a D2 dopamine antagonist, but not by D1 or alpha-adrenergic antagonists. Light-induced pigment dispersion in whole RPE-retinas is also blocked by a D2 dopamine antagonist.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547907 TI - Effect of halide ions on t-[35S]butylbicyclophosphorothionate binding. AB - The binding of t-[35S]butylbicyclophosphorothionate [( 35S]TBPS) to a site on the GABAA receptor complex is ion dependent. This study was conducted to determine the effects of ion species and concentration on the time course, affinity, and number of sites of [35S]TBPS binding. At a concentration of 200 mM ion, the time to equilibrium for [35S]TBPS binding was shortest for I-, followed by Br- less than Cl- less than F-. A similar rank order was observed for the concentration of ion required to produce half-maximal [35S]TBPS binding. Saturation binding experiments were conducted to evaluate the effect of increasing ion concentration on the KD and Bmax of [35S]TBPS binding. The Bmax was independent of both ion species and concentration. The receptor affinity, however, increased with increasing concentration for each ion. Calculated maximal affinity values were not different between ions; however, the EC50 to produce those values was different among ions and ranked in the same order as that for time course and maximal binding data. Association and dissociation rates for [35S]TBPS binding were greater in I- than in Cl-. These data emphasize the importance of ion selection and incubation times on [35S]TBPS binding. PMID- 2547906 TI - Phosphorylation of P400 protein by cyclic AMP-dependent protein kinase and Ca2+/calmodulin-dependent protein kinase II. AB - Purified P400 protein was phosphorylated by both purified Ca2+/calmodulin dependent protein kinase II (CaM kinase II) and the catalytic subunit of cyclic AMP-dependent protein kinase (A-kinase). Because P400 protein was suggested to function as an integral membrane protein, we investigated the phosphorylation of P400 protein using crude mitochondrial and microsomal fractions (P2/P3 fraction). Incubation of the P2/P3 fraction from mouse cerebellum with cyclic AMP or the catalytic subunit of A-kinase stimulated the phosphorylation of P400 protein. The phosphorylation of P400 protein was not observed in the P2/P3 fraction from mouse forebrain. Cyclic AMP and A-kinase enhanced the phosphorylation of several proteins, including P400 protein, suggesting that P400 protein is one of the best substrates for A-kinase in the P2/P3 fraction. Although endogenous and exogenous CaM kinase II stimulated the phosphorylation of some proteins in the P2/P3 fraction, the phosphorylation of P400 protein was weak. Immunoprecipitation with the monoclonal antibody to P400 protein confirmed that the P400 protein itself was definitely phosphorylated by the catalytic subunit of A-kinase and CaM kinase II. A-kinase phosphorylated only the seryl residue in P400 protein. Immunoblot analysis of the cells in primary culture of mouse cerebellum confirmed the expression of P400 protein, which migrated at the same position on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as that in the P2/P3 fraction. Incubation of the cultured cerebellar cells with [32P]orthophosphate resulted in the labeling of P400 protein.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547908 TI - Differential effects of iodide and chloride on allosteric interactions of the GABAA receptor. AB - t-[35S]Butylbicyclophosphorothionate [( 35S]TBPS) has been shown to bind to the GABAA receptor complex. The binding is modulated allosterically by drugs that interact at components of the receptor complex. The present studies were designed to evaluate the influence of ionic environment and state of equilibrium on the allosteric modification of [35S]TBPS binding. In both I- and Cl- under nonequilibrium conditions, diazepam, gamma-aminobutyric acid (GABA), and pentobarbital (PB) stimulate and methyl 6,7-dimethoxy-4-ethyl-beta-carboline-3 carboxylate (DMCM) inhibits [35S]TBPS binding. In addition, there is an inhibitory component to the effect of GABA and PB at higher drug concentrations. These effects are blocked by the appropriate antagonists for each drug. In Cl-, the stimulation of [35S]TBPS binding by drugs disappears at equilibrium, whereas the inhibition by GABA and PB persists. The inhibitory effect of DMCM in Cl- also disappears at equilibrium. When assayed in I- at equilibrium, however, DMCM stimulates [35S]TBPS binding. In addition, bicuculline, which is without effect under nonequilibrium conditions in either Cl- or I-, stimulates [35S]TBPS binding in I- at equilibrium. The persistent effects of DMCM, bicuculline, and GABA in I- are accompanied by alterations in the affinity of [35S]TBPS for its receptor. In addition, the stimulation of [35S]TBPS binding by GABA is associated with a decreased number of [35S]TBPS binding sites. These data demonstrate that receptor complex interactions with anions influence the responsiveness to drug binding. PMID- 2547909 TI - Reduced protein kinase C activity in ischemic spinal cord. AB - Protein phosphorylation was evaluated in a rabbit spinal cord ischemia model under conditions where cyclic AMP-dependent protein kinase (PK-A) and calcium/phospholipid-dependent protein kinase (PK-C) were activated. One hour of ischemia did not affect PK-A activity significantly; however, PK-C activity was reduced by more than 60%. In vitro phosphorylation of endogenous proteins by endogenous PK-C revealed that eight particulate and five cytosolic proteins showed stimulated phosphorylation by PK-C activators in control tissue, although this stimulation was virtually absent in ischemic samples. When control and ischemic particulate fractions were combined, the endogenous protein phosphorylation pattern under PK-C-activating conditions was similar to the ischemic sample, which suggests that inhibitory molecules may be present in the ischemic particulate fraction. In vitro phosphorylation of endogenous proteins under PK-A-activating conditions in ischemic tissue was similar to that in control tissue. The results suggest that the PK-C phosphorylation system is selectively impaired in ischemic spinal cord. In addition to reduced PK-C dependent phosphorylation, an Mr 64,000 protein was phosphorylated in ischemic cytosolic samples, but not in control samples. The phosphorylation of the Mr 64,000 protein was neither PK-C-dependent nor PK-A-dependent. These altered phosphorylation reactions may play critical roles in neuronal death during the course of ischemia. PMID- 2547910 TI - Catecholamine-sensitive adenylate cyclase in the white perch (Roccus americanus) retina: evidence for beta-adrenergic and dopamine receptors linked to adenylate cyclase. AB - We have examined the catecholamine-sensitive adenylate cyclase in the retina of the white perch (Roccus americanus). Both dopamine and the beta-adrenergic agonist isoproterenol stimulate cyclic AMP accumulation in this retina, but serotonin, an indoleamine, and phenylephrine, an alpha-adrenergic agonist, had no effect. The stimulation of adenylate cyclase by isoproterenol is more potent and effective than that of dopamine. The effects of dopamine and isoproterenol are mediated via independent dopamine and beta-adrenergic receptors. Haloperidol, a dopamine antagonist, blocks the stimulatory effect of dopamine but not of isoproterenol. Conversely, propranolol, a beta-adrenergic antagonist, blocks the stimulatory effect of isoproterenol but not of dopamine. The effects of dopamine and isoproterenol are not additive. In fractions of purified horizontal cells we found evidence for dopamine receptors linked to adenylate cyclase but did not find evidence for the presence of cyclase coupled beta-adrenergic receptors. The cellular location of the beta-adrenergic receptors is unknown. Our findings demonstrate the existence of both beta-adrenergic and dopamine receptors coupled to adenylate cyclase in the white perch retina. However, we did not find either epinephrine or norepinephrine, endogenous ligands of the beta-receptor, to be present in retinal extracts subjected to HPLC. PMID- 2547911 TI - In vivo modulation of the N-methyl-D-aspartate receptor complex by D-serine: potentiation of ongoing neuronal activity as evidenced by increased cerebellar cyclic GMP. AB - Direct intracerebellar injections of N-methyl-D-aspartate (NMDA) or D-serine elicited dose-dependent increases in cerebellar cyclic GMP levels, in vivo in the mouse. The actions of D-serine were antagonized by the competitive NMDA receptor antagonist 3-(2-carboxypiperazin-4-yl) propyl-1-phosphonic acid and by the phencyclidine receptor agonist MK-801, observations supporting actions at the NMDA-coupled glycine receptor. In addition, the actions of D-serine were antagonized by a partial agonist (D-cycloserine) and an antagonist (HA-966) of the NMDA-coupled glycine receptor. These data are all consistent with D-serine acting at the NMDA-coupled glycine receptor and represent the first demonstration of glycine receptor potentiation of ongoing NMDA-mediated neuronal activity in the CNS, rather than potentiation of exogenous NMDA. PMID- 2547912 TI - gamma-Aminobutyric acidB receptor activation modifies agonist binding to beta adrenergic receptors in rat brain cerebral cortex. AB - The interaction of isoproterenol with beta-adrenergic receptor (beta AR) binding sites was measured in membranes prepared from rat brain cerebral cortical slices previously incubated in the presence or absence of gamma-aminobutyric acid (GABA) receptor agonists. Both GABA and baclofen, but not isoguvacine, altered beta AR agonist binding by increasing the affinity of both the low- and high-affinity binding sites and by increasing the proportion of low-affinity receptors. The response to baclofen was stereoselective, and the effect of GABA was not inhibited by bicuculline. The results suggest that GABAB, but not GABAA, receptor activation modifies the coupling between beta AR and stimulatory guanine nucleotide-binding protein, which may in part explain the ability of baclofen to augment isoproterenol-stimulated cyclic AMP accumulation in brain slices. PMID- 2547913 TI - Partial deficiency of complexes I and IV of the mitochondrial respiratory chain in skeletal muscle of two patients with mitochondrial myopathy. AB - Respiratory chain enzymes were studied in isolated mitochondria of two patients with mitochondrial myopathy. Both patients had been suffering from chronic progressive external ophthalmoplegia and abnormal muscular fatigability since late childhood. One of the patients exhibited the complete triad of symptoms characteristic of Kearns-Sayre syndrome. Venous lactate levels at rest and during minimal exercise were increased in both patients. Histochemical examination of muscle revealed ragged red fibres and intermingled fibres negative for cytochrome c oxidase. Biochemical studies showed decreased activities of complex I and complex IV of the respiratory chain in both patients. Reduced minus oxidized spectra of mitochondrial cytochromes revealed a decreased content of cytochrome aa3 in only one patient, but a normal content in the other. A combined deficiency of complexes I and IV in muscle might either be due to a deficiency of a single subunit common to both complexes or to a coincidental deficiency of both complexes expressed either in the same or in different fibres. PMID- 2547914 TI - Japanese encephalitis in the Kurume region of Japan: CT and MRI findings. AB - Neurological, computed tomography (CT) and magnetic resonance imaging (MRI) findings were recorded from 13 patients with Japanese encephalitis (JE) in the Kurume region diagnosed by serological criteria. The patients averaged 63 years of age, and 5 were older than 70 years. The serological data mostly indicated a primary response. Hemiplegia and tetraplegia were common, together with extrapyramidal signs. A few cases had a stroke-like onset and cerebral haemorrhage during the course of JE. CT and MRI in 7 cases revealed abnormalities in the thalamus and basal ganglia including the putamen. The CT and MRI findings from the acute stage to the convalescent stage were considered to be characteristic of JE. PMID- 2547915 TI - An epidemiologic investigation of forced expiratory volume at 1 second and respiratory symptoms among employees of a toluene diisocyanate production plant. AB - Pulmonary function tests were done and compared to current and past potential exposure levels of toluene diisocyanate (TDI) for 57 TDI manufacturing workers and 89 workers not exposed to TDI or other known respiratory hazards. The average TDI plant experience was 4.1 years (standard deviation = 2.8). Routine industrial hygiene measurements have shown TDI exposure below a time-weighted average of 0.005 parts per million and a short-term exposure level of 0.02 parts per million. A certified industrial hygienist ranked department and job classification by level of potential exposure to TDI (none, low, moderate, and high). A questionnaire was administered to determine the prevalence of respiratory symptoms and smoking habits. Using backward regression analysis, cumulative pack-years of cigarette smoking and prevalence of lower respiratory symptoms were statistically significant predictors of a standardized forced expiratory volume at 1 second observed v expected difference; however, TDI exposure, whether classified as current, highest career level, cumulative, or cumulative highest-to-date, was not associated with a decline in forced expiratory volume. PMID- 2547916 TI - Effect of fish oil on blood pressure and serum lipids in hypertension and hyperlipidaemia. AB - We studied the antihypertensive and hypolipidaemic effects of fish oil containing eico-sapentaenoic acid and docosahexaenoic acid in a capsule preparation in patients with mild to moderate essential hypertension and in patients with hypercholesterolaemia. In addition, we used two independent procedures to analyse changes in blood pressure, casual and self-recorded blood pressure measurements. A very moderate blood pressure lowering effect of fish oil was confirmed in this study, and a slight antihyperlipidaemic effect in plasma triglycerides was demonstrated. During the fish oil treatment, casual blood pressure values were consistently lower than self-recorded values. It is assumed that this was an observer error due to knowledge of the treatment. PMID- 2547917 TI - Disparities in blood pressure control under various antihypertensive regimens. AB - Ambulatory blood pressure recordings and stress blood pressures during exercise were compared among hypertensive patients effectively treated with oxprenolol, nitrendipine, enalapril or low-dose clonidine. After 6 months of therapy, the means of blood pressure at rest and casual diastolic pressure were nearly identical among the four therapeutic groups. Although all pressures fell to within the normotensive range, casual systolic pressures were lower in patients treated with sympatholytic agents than in those taking enalapril. In contrast, average ambulatory blood pressure was less controlled in patients given clonidine or enalapril than in those given oxprenolol or nitrendipine. During physical stress patients taking clonidine showed the highest stress blood pressures and those taking oxprenolol the lowest pressures. The study demonstrated that although blood pressure was reduced to within the normotensive range in all four therapeutic groups, analysis of values of ambulatory blood pressure and stress blood pressure during physical activity showed a disparate pattern of antihypertensive efficacy. PMID- 2547918 TI - Oral synovial sarcoma: report of a case. AB - A case of synovial sarcoma arising in the buccal mucosa of a 36-year-old man is reported, and the literature is reviewed. Immunohistochemical findings concerning the histogenesis of the tumor are discussed. PMID- 2547919 TI - Temporomandibular joint ankylosis: surgical treatment and long-term results. AB - The surgical treatment of 26 patients with ankylosis of the temporomandibular joint, as well as various methods and materials used for functional restoration are described. The significance of radical removal of the ankylotic bone, as well as the advantages of the interpositioning of the lyophilized cartilage, are emphasized. PMID- 2547920 TI - Nonporous hydroxylapatite in the repair of alveolar clefts in a primate model: clinical and histologic findings. AB - Twelve adult Rhesus monkeys with surgically created alveolar clefts (ACs) underwent repair that used nonporous hydroxylapatite (NPHA) granules on one side and autogenous particulate bone in the contralateral defect. Clinical and radiographic evaluations at 1, 2, 3, 6, and 12 months postoperatively disclosed some displacement of granules initially, but this stabilized by 2 months. Three of 12 sites repaired with NPHA and four of 12 sites repaired with bone dehisced; however, all of these healed by 2 months. No untoward inflammatory or resorptive changes were observed beyond 3 months. The repaired alveolar processes were similar in bulk and contour in sites grafted with NPHA and with bone. Histologically, giant cells were noted at 3 months, but the number decreased at 6 months and remained at that level up to 12 months. The NPHA granules were embedded in mature fibrous connective tissue with osseous ingrowth evident in eight of the 12 defects. It was concluded that NPHA granules are an acceptable implant material for repair of residual alveolar clefts in which erupting teeth and orthodontic movements are not factors. PMID- 2547921 TI - Bone induction in a composite allogeneic bone/alloplastic implant. AB - This study evaluated the morphogenetic properties of the bone matrix as a biological delivery system in a composite allogeneic bone/alloplastic implant placed in extraskeletal recipient sites in rats. Seventy-two allogeneic diaphyseal bone cylinders were chemosterilized to obtain autolyzed antigen extracted allogeneic (AAA) bone and implanted in twenty 140- to 160-day-old rats. In 30 implants, porous hydroxyapatite biomatrix (HAB) was inserted into the AAA bone cylinders creating the composite implants. Serial sections from the specimens harvested at 23, 30, 45, 53, and 63 days showed that 84% of the AAA bone cylinders induced the differentiation of variable amounts of endochondral bone and that in 61% bone formed in the interior of the cylinders; 74% of the composite implants had no bone-inductive effect, recruiting instead macrophages and multinucleated giant cells at the HAB/mesenchymal interface. In all instances bone failed to form within the porous HAB or in proximity to the AAA bone matrix facing the interior of the cylinders. When present, bone formed only in localized areas within the matrix. Histologic analysis showed that high deposits of induced bone are correlated to high levels of matrix vascular invasion and mesenchymal cell aggregation. Within the limits of this study on rodents, the results suggest that the HAB inhibits and delays the local differentiation of induced bone in the composite implants. PMID- 2547923 TI - Effect of dexamethasone on herpes simplex virus replication in mouse neuroblastoma cells (NB41A3): receptor characteristics. AB - Monolayers of NB41A3 (MNB) cells were exposed to pharmacologic doses of dexamethasone (DXM). After 24-h, the cells were infected with herpes simplex virus type 1 (HSV-1) at a multiplicity of infection (m.o.i.) = 0.1. After every 24-h period, extracellular and intracellular virus aliquots were collected and frozen. The aliquots were titered in a standard plaque-forming assay. It was shown that the hormone led to a statistically significant increase of extra- and intracellular virus titers above the titers exhibited by these cells without added hormone. The same experiment was repeated in Vero cells, but the hormone did not elevate the resultant HSV-1 titers. A binding assay was performed on these two cell lines by use of 3H-DXM to determine if a DXM receptor was present. Specific binding was seen, but only in the MNB cell line. The Bmax of this receptor was 480 fmol/mg protein and it had a Kd of 2.3 nM. The S value of the receptor ligand complex equalled 8.0. These results indicate that cells possessing hormone receptors allow for a more efficient replication of the virus and suggest that these hormones may play an important role in the exacerbation of herpes simplex virus infection in vivo. PMID- 2547922 TI - Myocardial potassium balance during adrenergic stimulation. AB - 1) The primary receptor mechanism of catecholamine-induced myocardial potassium uptake is beta 1-adrenoceptor stimulation. Thus, K+ uptake seems to be a general effect of beta-adrenergic stimulation, dominated by beta 1-receptors in heart and by beta 2-receptors in skeletal muscle according to subtype preponderance in either tissue. In the myocardium there is also an effect of alpha 1-adrenoceptor stimulation which causes a significantly smaller uptake and requires higher catecholamine concentrations. 2) Both humoral and nervous adrenergic stimulation of the heart induce a significant potassium uptake which transiently reduces coronary sinus K+ concentration. It is likely that these changes affect cardiac functioning in vivo. During intense endogenous sympathetic activity and by high dose pharmacological interventions, the magnitude of change in coronary sinus concentration suggests that the reduction in extracellular K+ within the myocardium could be up to 1 mM. Under vulnerable conditions like hypokalemia and localized ischemia such changes might contribute to the risk for malignant arrhythmias. 3) Presumably net myocardial K+ accumulation is accompanied by a reciprocal reduction of intracellular Na+ concentration, which tends to reduce myocardial contractility and contribute to impaired cardiac function after a period of strong adrenergic stimulation. In vivo the negative inotropic effect could not be detected as long as catecholamines were supplied, but it occurred after stimulation was stopped. 4) In the intact beating heart beta-adrenergic stimulation increases Na,K-pumping 2.5 fold, from 15% of the maximum possible pump rate in control to 40% of maximum at high inotropy. These findings imply the presence of a substantial spare Na,K-pump capacity of the non-ischemic myocardium, even during intense sympathetic activity. Comparison of changes in pump rate and accumulated ionic shifts indicates that catecholamine-induced stimulation of Na,K-ATPase might be due to increased sensitivity for intracellular sodium. PMID- 2547924 TI - Chemoattractant activity of Entamoeba histolytica for human polymorphonuclear neutrophils. AB - The ability of axenic Entamoeba histolytica trophozoites and amoebic protein preparations to stimulate chemotaxis of human polymorphonuclear neutrophils (PMN) was evaluated. Virulent E. histolytica (strain HM1:IMSS) stimulated chemotaxis (delta distance = 0.55 +/- 0.02 mm, P less than 0.01 vs. control medium). Sonicated (100 micrograms protein/ml) or homogenized (500 micrograms protein/ml) virulent amoebae also significantly stimulated PMN chemotaxis, whereas preparations of the nonpathogenic "Entamoeba-like" Laredo strain did not stimulate chemotaxis. Preparations of subcellular fractions of E. histolytica demonstrated maximal stimulation of PMN chemotaxis existed in nonvesiculated membranes and the supernatant from plasma membranes. PMID- 2547925 TI - The topography of acetylcholinesterase in dimyristoylphosphatidylcholine liposomes. AB - Unilamellar liposomes prepared from sn-3-(dimyristoyl)phosphatidylcholine (DMPC) in the presence and absence of acetylcholinesterase were examined by ESR for lipid/protein interactions. Using 5-, 12-, and 16-doxyl stearic acid probes incorporated into the phospholipid bilayers, no measurable differences in the gel to liquid-crystalline phase transition temperature of DMPC (as determined by ESR spectroscopy) were observed when the enzyme was present. These results have established that no significant incorporation of acetylcholinesterase into the hydrophobic region of the phospholipid bilayer is detectable at the protein: lipid ratios used in these experiments. Confirmation of these results was also obtained by differential scanning calorimetry. Interaction of the enzyme with the outermost region of the bilayer was established by trypsin digestion which indicated that as much as 25% of liposome-associated enzyme was removable and was, therefore, exposed to the outer surface. The results of this study have established that acetylcholinesterase associated with unilamellar DMPC liposomes was primarily entrapped within the aqueous compartment of the vesicles and was not present in the phospholipid bilayer. PMID- 2547928 TI - Anaerobic infections. PMID- 2547927 TI - Synthesis of 1,5-diaryl-3-methyl-1H-pyrazolo[4,5-c]isoquinolines and studies of binding to specific peripheral benzodiazepine binding sites. AB - Some 1,5-diaryl-3-methyl-1H-pyrazolo[4,5-c]isoquinolines were synthesized and tested for their ability to displace [3H]clonazepam or [3H]Ro 5-4864 from their specific binding on the central and peripheral benzodiazepine receptors. None of the tested compounds showed any activity as central binding inhibitors, while most of them were specific as peripheral binding inhibitors, although they were not very potent. PMID- 2547926 TI - Kappa-opioid agonists do not inhibit adenylate cyclase. AB - Morphine, levorphanol and D-Ala2, Met enkephalin, but not bremazocine, pentazocine, U-50 488H and U-69 593, were found to inhibit adenylate cyclase activity dose-dependently in bullfrog brain membranes. The inhibition of the enzyme activity was abolished by naloxone. These results suggest that the signalling transduction of mu- and delta-agonists is partially mediated by the adenylate cyclase system coupled with opioid receptors, but that kappa-receptors may not be associated with adenylate cyclase. PMID- 2547929 TI - Cyclic guanosine 3',5' monophosphate mediates the inhibitory effect of atrial natriuretic factor in adrenergic, neuronal pheochromocytoma cells. AB - This study tests the hypothesis that atrial natriuretic factor (ANF) inhibits catecholamine release from rat pheochromocytoma cells by increasing levels of intracellular cyclic GMP (cGMP). Rat differentiated pheochromocytoma cells are a model of adrenergic nerves and allow the exploration of the effects of various hormones, autacoids, drugs and neuromodulators on adrenergic neurotransmission in cell culture. Synthetic rat ANF (99-126) inhibited K+-induced norepinephrine and dopamine release, as measured by high-performance liquid chromatography, in a concentration-dependent manner over the concentration range of 10(-11) to 10(-8) M. ANF stimulated intracellular cGMP accumulation, as measured by specific radioimmunoassay, in a concentration-dependent manner over the same concentration range. The cGMP analog, N2-2'-O-dibutyryl cGMP also inhibited K+-induced norepinephrine and dopamine release in a concentration-dependent manner. The results of this study are consistent with the hypothesis that ANF acts as an inhibitory neuromodulator in adrenergic nerves via the second messenger, cGMP. PMID- 2547930 TI - Adenosine-angiotensin II interactions. Part I. Role of adenosine in regulating angiotensin II-induced potentiation of noradrenergic neurotransmission and angiotensin II-induced vasoconstriction. AB - The pharmacology of adenosine and angiotensin II (AII) suggests that endogenous adenosine could function to regulate some of the biological effects of AII. The goal of this study was to test the hypothesis that endogenous adenosine inhibits the ability of AII to potentiate noradrenergic neurotransmission and/or to directly contract vascular smooth muscle. Two approaches were used to assess the physiological import of adenosine-AII interactions on neurotransmission and vascular tone. First, the effects of exogenous adenosine and AII, separately and in combination, on vascular responses to periarterial nerve stimulation (PNS) and vascular tone were determined. Second, the effects of an adenosine receptor antagonist, 1,3-dipropyl-8-p-sulfophenylxanthine, on AII-induced potentiation of vascular responses to PNS and on AII-induced direct vasoconstriction were examined. All studies were conducted in the in situ blood perfused rat mesentery. Intra-arterial and i.v. infusions of AII potentiated responses to PNS and increased vascular tone, whereas i.a. infusions of adenosine had the opposite effects. Intra-arterial infusions of adenosine potentiated the ability of i.a. AII to enhance responses to PNS; however, i.a. adenosine attenuated the direct vasoconstrictive action of i.a. AII. Intra-arterial infusions of 1,3-dipropyl-8-p sulfophenylxanthine, at a dose which antagonized the effects of exogenous adenosine, did not alter the effects of either i.a. AII or i.v. AII on vascular responses to PNS or on vascular tone. These results indicate that although adenosine has the potential to regulate AII-induced enhancement of noradrenergic neurotransmission and AII-induced direct vasoconstriction, such regulation does not occur under the conditions of this study. PMID- 2547931 TI - Behavioral pharmacological profile of CGS 19755, a competitive antagonist at N methyl-D-aspartate receptors. AB - CGS 19755 (cis-4-phosphonomethyl-2-piperidine-carboxylic acid), a competitive antagonist at N-methyl-D-aspartate (NMDA)-preferring receptors, blocked both NMDA induced convulsions in normal CF1 mice and sound-induced wild running in seizure prone DBA/2 mice. The ED50 values for CGS 19755 to produce these effects (in the range of 2 mg/kg i.p.) were at least 3-fold lower than those which impaired the traction reflex, an index of motor coordination. When administered p.o. by gavage, CGS 19755 had little or no effect in these test procedures. In an experimental model of anxiety in rats, CGS 19755 significantly increased conflict responding within a relatively narrow dose range (minimum effective dose, 1.73 mg/kg i.p.). At higher doses of CGS 19755, this effect appeared to be obscured by drug-induced reductions in overall responding. Potential muscle relaxant effects were also suggested by the generalization of CGS 19755 to diazepam discriminative stimuli (ED50 = 9.0 mg/kg i.p.) and by impaired rotorod performance (ED50 = 6.2 mg/kg i.p.) in rats. Although some resemblances were apparent between the behavioral effects of CGS 19755 and those of phencyclidine-type drugs, the phencyclidine-like behaviors appeared only at considerably higher doses of CGS 19755 than those associated with anticonflict activity, and only partial generalization of CGS 19755 to dexoxadrol was observed at high doses. CGS 19755 promises to be an important new research tool for investigating the function of brain NMDA receptors. PMID- 2547932 TI - Role of purines and xanthine oxidase in reperfusion injury in perfused rat liver. AB - The purpose of this study was to evaluate the possible involvement of xanthine and xanthine oxidase in reperfusion injury in a low-flow, reflow model of liver perfusion. Livers were perfused at flow rates around 25% of normal for 90 min and then at normal flow rates (4 ml/g/min) for 30 min. When flow was restored to normal, malondialdehyde and lactic dehydrogenase (LDH) were released into the effluent perfusate. Malondialdehyde production rapidly reached values of 300 nmol/g/hr whereas LDH increased from basal levels of 100 to 600 U/l upon reperfusion. Trypan blue was taken up exclusively in cells in pericentral regions of the liver lobule under these conditions. Xanthine and hypoxanthine in the effluent perfusate increased steadily during the low-flow period reaching values around 5 and 10 microM, respectively, and decreased rapidly after the flow was restored to normal. Perfusion with nitrogen-saturated buffer for 3 min before restoration of normal flow rates or infusion of the radical scavenger catechin (400 microM) reduced cell damage by about 50%. Infusion of allopurinol (2-6 mM), an inhibitor of xanthine oxidase, prevented reperfusion injury in a dose dependent manner. Taken together, these data indicate that a reperfusion injury occurs in liver upon reintroduction of oxygen which is initiated by oxidation of xanthine and hypoxanthine via xanthine oxidase and ultimately leads to production of lipid peroxides. Surprisingly, infusion of xanthine (4 mM), substrate for xanthine oxidase, reduced hepatocellular injury on reperfusion. LDH release was decreased from values around 700 to less than 200 U/l and trypan blue uptake in pericentral region was prevented totally by xanthine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547933 TI - A role for histamine and H2-receptors in opioid antinociception. AB - To investigate the role of brain H2-receptors in opioid antinociceptive mechanisms, the effects of several antagonists of histamine H2-receptors were determined on morphine (MOR)-induced antinociception, opioid-mediated footshock induced antinociception (FSIA) and on other opiate effects in rats. Zolantidine dimaleate (ZOL), the first brain-penetrating H2 antagonist (0.03-1.6 mumol/kg s.c.) caused a dose-related inhibition of MOR antinociception in both the tail flick and hot-plate tests, with no effect on base-line responding. ZOL also inhibited opioid FISA with a similar potency. MOR-induced locomotor activity was also reduced by ZOL, but no effect was seen on MOR-induced hyperthermia, catalepsy or lethality. ZOL (10(-5) M) was inactive at mu, delta or kappa opioid receptors and showed at least 35-fold higher affinity at the H2-receptor than at receptors for serotonin, dopamine, norepinephrine or acetylcholine in brain. To clarify further the role of H2-receptors in ZOL's antiopiate activity, the potencies of seven structural congeners of ZOL were determined on the H2-receptor and on MOR antinociception. Over 3 orders of magnitude, the rank order of potencies of the compounds for inhibiting MOR antinociception was highly correlated with their potencies as H2 antagonists. Cimetidine, unlike other H2 antagonists, potentiated MOR antinociception, potentiated opioid FSIA and increased brain MOR levels, actions that are not likely to be due to blockade of H2-receptors. These findings strongly suggest that stimulation of opioid receptors leads to antinociception by mechanisms that include activation of brain H2-receptors. PMID- 2547934 TI - Analysis of the presence of postjunctional alpha-2 adrenoceptors in the rat anococcygeus muscle. AB - When the tone is raised by guanethidine, rat anococcygeus muscle produces inhibitory responses to field stimulation, whose mechanism is not understood properly. The present study is an attempt to investigate the role of alpha adrenoceptors in the field stimulation-induced relaxations in isolated rat anococcygeus muscle. When the tissues are contracted with clonidine, UK-14,304 and low doses of oxymetazoline, field stimulation produced relaxations at lower frequencies, but not in the tissues precontracted with phenylephrine and norepinephrine. Relaxations induced by low frequencies were blocked by idazoxan, but not by phentolamine, prazosin, indomethacin, N-methyl-hydroxylamine, ouabain or 3,4-diminopyridine. When the tone of the muscle is raised by norepinephrine, prazosin reversed the field stimulation-induced contractions to relaxation responses. The data of the present study suggested the possible involvement of alpha-2 adrenoceptors during the field stimulation-induced relaxations of the rat anococcygeus muscle. To analyze and quantitate the alpha-2 adrenoceptor antagonism in the rat anococgygeus muscle, Schild analyses of clonidine-induced contractions against idazoxan were conducted either for idazoxan alone or after partially alkylating the alpha-1 adrenoceptors with phenoxybenzamine and by pharmacologic resultant analysis by blocking the alpha-1 adrenoceptors with prazosin. The Schild regression for idazoxan and pharmacologic resultant analysis suggested that the rat anococcygeus muscle responds to alpha-2 agonists with alpha-1-mediated contractions and idazoxan competes with alpha-1 antagonists for the same site, i.e., alpha-1 adrenoceptor site. However, the atypical Schild regression of idazoxan after partial alkylation with phenoxybenzamine indicated the existence of a second alpha adrenoceptor site in the rat anococcygeus muscle.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547935 TI - Serotonergic involvement in the antinociceptive action of and the development of tolerance to the kappa-opioid receptor agonist, U-50, 488H. AB - The antinociceptive action of U-50,488H [( trans-(+/-)-3,4-dichloro-N-methyl-N-[2 (1-pyrrolidinyl)- cyclohexyl]benzeneacetamide]), a selective kappa-opioid receptor agonist, was antagonized by serotonergic (5-HT) but not by noradrenergic receptor antagonists. U-50,488H (i.c.v.) was antagonized by more than 2-fold by i.c.v. administered pindolol, methysergide, mianserin, ketanserin or pirenperone. A similar degree of antagonism of U-50,488H (i.c.v.) was found after intrathecal (i.t.) treatments with pindolol or methysergide but not with mianserin, ketanserin or pirenperone. When U-50,488H was given i.t., its antinociceptive action was antagonized by pindolol or methysergide administered i.t. but potentiated by mianserin, ketanserin or pirenperone administered i.t. Tolerance to the antinociceptive action of U-50,488H was induced in mice using slow release preparations of U-50,488H. A smaller degree of cross-tolerance to the antinociceptive action of 5-HT also developed. 5-HT receptor antagonists (pindolol or ketanserin) were coadministered with U-50,488H to test for their effect on the development of tolerance to U-50,488H. A greater degree of tolerance was observed after ketanserin administration whereas the development of tolerance was partially blocked by the coadministration of pindolol. Changes in the properties of the kappa-opioid, 5-HT1 and 5-HT2 receptor binding sites in the cortical, striatal and spinal tissues of control and U-50,488H-tolerant mice were investigated. There were no significant changes in the maximum binding values of [3H]ethylketazocine, [3H]-5-HT or [3H]ketanserin. The Kd values of all ligands increased approximately 2-fold in most of the regions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547937 TI - Effects of ethanol on muscarinic receptor-stimulated phosphoinositide metabolism during brain development. AB - The pattern of muscarinic receptor-stimulated inositol metabolism during postnatal development has a striking resemblance with the curve of rat brain growth spurt. Therefore, the enhanced hydrolysis of membrane inositol lipids by cholinergic agonists during this period may have a relevant role in cell proliferation and differentiation. In this study we have investigated whether exposure to EtOH to rat pups during the brain growth spurt, known to cause a permanent microencephaly, would alter muscarinic receptor-stimulated inositol metabolism in cerebral cortex. Female Long-Evans rats were administered 4 g/kg of EtOH, in two doses of 2 g/kg, by gastric intubation from postnatal day 4 to day 10. This treatment did not have any effect on the pups' body weight as compared to an equally handled, sucrose-fed group of animals. Blood EtOH concentration in the pups during exposure ranged between 237 and 283 mg/dl. Muscarinic receptor stimulated inositol metabolism was measured in cerebral cortex slices of control and EtOH-treated rats at days 4, 7, 10, 12, 20 and 45 of age. Carbachol-induced accumulation of [3H] inositol phosphates was reduced significantly in EtOH exposed animals on day 7 and 10, but not at other ages. This decrease was not due to an alteration of muscarinic receptor density or affinity. Exposure to EtOH had no effect on phosphoinositide metabolism stimulated by norepinephrine, serotonin or histamine in cerebral cortex, whereas the effect of glutamate was increased. Similar results were observed in the hippocampus. An identical treatment with EtOH in adult rats did not cause alteration in any of the biochemical parameters of the cholinergic system measured.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547936 TI - Characterization of the effects of the acute and chronic administration of phencyclidine on the release of adrenocorticotropin, corticosterone and prolactin in the rat: evidence for the differential development of tolerance. AB - Phencyclidine (PCP) is a widely used drug of abuse; however, little is known of its effects on neuroendocrine function. The present study characterized the effects of the acute and chronic administration of PCP on the release of adrenocorticotropin, corticosterone and prolactin in the rat. For the acute studies, PCP hydrochloride (0.5-10.0 mg/kg s.c.) was administered and the subjects were sacrificed 15 to 300 min later. The acute administration of PCP produced rapid and long-lasting increases in plasma levels of adrenocorticotropin and corticosterone but decreased plasma levels of prolactin. For the chronic studies, PCP (1.0-20.0 mg/kg/day s.c.) was injected daily and the subjects sacrificed 60 min after injection on day 15. PCP continued to stimulate the pituitary-adrenal axis after chronic administration; however, there was a decrease in the magnitude of response, indicating the development of some degree of tolerance. In contrast, none of the doses of PCP tested decreased plasma prolactin levels in chronically treated subjects. There were no differences in plasma or brain levels of PCP in the chronically PCP-treated rats, indicating that tolerance was not due to changes in the biodisposition of PCP. These results indicate that PCP disrupts neuroendocrine function markedly in the rat. The differential development of tolerance to the effects of PCP on the pituitary adrenal axis and prolactin release may indicate that different neurochemical substrates underlie the effects of PCP on different endocrine systems. PMID- 2547938 TI - Cellular mechanisms of endothelin in rabbit aorta. AB - The present studies were designed to investigate the cellular actions of endothelin in rabbit aorta. Endothelin produced concentration-dependent contraction of rabbit isolated aortic rings which was independent of the endothelium; the EC50 values were 6.1 and 5.6 nM for endothelium-intact and endothelium-denuded vascular rings, respectively. Endothelin (1 nM) did not induce the release or inhibit the effect of endothelium-derived relaxing factor in precontracted aortic rings. Removal of calcium from the external bathing medium reduced the maximal contractile response induced by endothelin (0.1 microM) by only 12%; however, addition of ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid (0.1 mM) to the calcium-free medium produced a marked inhibition (approximately 75%) of endothelin-induced contraction. The dihydropyridine calcium channel antagonists nicardipine (1 microM) or nifedipine (1 microM), and also diltiazem (1 microM), had little or no effect on endothelin concentration-response curves. Contraction produced by endothelin (30 nM) was not associated with an alteration in the levels of cyclic 3',5'-adenosine monophosphate or cyclic 3',5-guanosine monophosphate in either endothelium-intact or endothelium-denuded aortic rings. Endothelin (1 nM-0.1 microM) produced concentration-dependent stimulation of phosphatidylinositol (Pl) turnover in aortic rings when exposed to tissues for periods greater than or equal to 15 min. Endothelin-induced stimulation of Pl turnover was unaffected by nicardipine (0.1 microM). In calcium-free medium (+0.1 mM ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid) basal Pl turnover was reduced; however, endothelin (1 nM-0.1 microM) produced similar percentage increases over basal levels to those observed in the presence of calcium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547940 TI - An assessment of the effects of glucocorticoids on degranulation, chemotaxis, binding to vascular endothelium and formation of leukotriene B4 by purified human neutrophils. AB - Glucocorticoids exert their actions through a time-dependent, receptor-mediated, protein synthesis- and RNA synthesis-dependent mechanism. We have assessed the effects of 24-h culture of human neutrophils with dexamethasone on degranulation, chemotaxis, binding to vascular endothelium and formation of leukotriene B4. Purified neutrophils contained an average of 2896 [3H]dexamethasone binding sites per cell with a Kd of 4.1 X 10(-9) M for [3H]dexamethasone binding. Cells exposed to dexamethasone (10(-6) M) released equal or greater quantities of the lysosomal enzymes, lysozyme and beta-glucuronidase in response to formylmethionyl-leucyl phenylalanine, serum activated zymosan, and the tumor promoting phorbol diester 12-O-tetradecanoylphorbol-13-acetate compared to controls. Culture with dexamethasone also did not inhibit neutrophil chemotaxis in response to a range of concentrations of formylmethionyl-leucyl-phenylalanine, or did it inhibit binding of neutrophils to cultured endothelial cells stimulated by either leukocyte activators (formylmethionyl-leucyl-phenylalanine and platelet activating factor) or endothelial activators (interleukin-1, lipopolysaccharide or 12-O-tetradecanoylphorbol-13-acetate). Spontaneous adherence of neutrophils to endothelial cells was inhibited (82.9 +/- 6.8% of control, P less than .025, n = 18). Neither in vitro or in vivo glucocorticoids inhibited neutrophil leukotriene B4 formation induced by either the calcium ionophore A23187 or serum activated zymosan. We conclude that human neutrophils are not functionally inactivated by glucocorticoids and suggest that the mechanism by which glucocorticoids inhibit neutrophil accumulation at inflammatory sites may be by inhibition of the production of chemoattractants and endothelial activators rather than inhibition of their actions. PMID- 2547939 TI - Characterization of the striatal M2 muscarinic receptor mediating inhibition of cyclic AMP using selective antagonists: a comparison with the brainstem M2 receptor. AB - Pharmacological profiles of the striatal and brainstem M2 receptors were developed with a group of selective muscarinic antagonists. The striatal M2 muscarinic receptor was identified by its inhibition of [3H]cyclic AMP levels, whereas the brainstem M2 receptor was characterized using competition with [3H]quinuclidinyl benzilate binding. The potency of pirenzepine does not differentiate clearly between the striatal M2 receptor (Ki approximately 300 nM) and the brainstem M2 receptor (Ki = 219 nM) or peripheral M2 receptors. In the present study, we used 4-diphenylacetoxy-N-methylpiperidine methbromide, hexahydrosiladifenidol, AF-DX 116 and methoctramine to characterize the striatal and brainstem M2 receptors in more pharmacological detail. For comparison, the potencies of these antagonists were also measured at cortical M2 receptors (using competition with [3H]pirenzepine binding). The potencies of 4-diphenylacetoxy-N methylpiperidine methbromide (KB = 0.19 nM) and hexahydrosiladifenidol (KB = 14 nM) in blocking the striatal M2 receptor suggested similarity to those M2 receptors localized in certain smooth muscles or in glands. However, AF-DX 116 (KB = 155 nM) and methoctramine (KB = 47 nM) were considerably more potent in blocking the striatal M2 receptor than as reported in functional studies in smooth muscle or glands. Thus, the profile of the striatal M2 receptor obtained with these antagonists did not match in all respects with either glandular (probable M4 gene product) or cardiac (probable M2 gene product) muscarinic receptors. In contrast, our data with the brainstem M2 receptor was highly correlated (r = 0.93) with literature data regarding the cardiac muscarinic system.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547941 TI - SCH 39370, a neutral metalloendopeptidase inhibitor, potentiates biological responses to atrial natriuretic factor and lowers blood pressure in desoxycorticosterone acetate-sodium hypertensive rats. AB - SCH 39370 (N-[N-[1-(S)-carboxyl-3-phenylpropyl]-(S)-phenyl-alanyl]-(S)-isoserin e) is a potent and specific inhibitor of neutral metalloendopeptidase (NEP) from rabbit kidney (IC50 = 11.2 +/- 1.9 nM) and is devoid of angiotensin-converting enzyme inhibitory activity at 1 microM. We evaluated the effect of NEP inhibition with SCH 39370 on the inactivation of atrial natriuretic factor (ANF) and on cardiovascular function in rats. SCH 39370 effectively prevented in vitro degradation of ANF (99-126) by a purified rabbit kidney NEP. SCH 39370 (30 mg/kg s.c) significantly delayed the disappearance of immunoreactive (ir) ANF from plasma in rats after an i.v. infusion of ANF (1 microgram/kg/min for 30 min): the plasma ir ANF level at 15 min postinfusion was 1.5 +/- 0.3 ng/ml vs. 0.3 +/- 0.04 ng/ml in the control. SCH 39370 also delayed the disappearance of ir ANF after infusion of the peptide (0.1 microgram/kg/min for 30 min) which increased plasma levels to those observed during volume expansion. This effect was accentuated markedly in rats with bilateral nephrectomy. The hypotensive response to injection of ANF (30 micrograms/kg i.v.) in spontaneously hypertensive rats (-38 +/- 6 mm Hg vs. -13 +/- 2 mm Hg in the control) and the diuretic and natriuretic responses to ANF in normal rats were potentiated by SCH 39370 (30 mg/kg s.c.), respectively. The results suggest that NEP can play a role in ANF disposition in vivo and that potentiation of the biological activities of high doses of ANF by SCH 39370 may be consequent to its inhibitory effect on ANF degradation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547942 TI - Release of gamma-[3H]aminobutyric acid (GABA) from electrically stimulated rat cortical slices and its modulation by GABAB autoreceptors. AB - Slices of rat temporo-parietal cortex were prelabeled with gamma-[3H]aminobutyric acid ([3H]GABA), in the presence of the glial GABA uptake inhibitor beta-alanine. The slices were then superfused with a medium containing the GABA transaminase inhibitor aminooxyacetic acid and stimulated electrically (5 min, 2 msec, 36 mA at 5 or 10 Hz), in the presence of the neuronal GABA reuptake inhibitor SK&F 89976A [N-(4,4-diphenyl-3-butenyl)-nipecotic acid] and of beta-alanine. Representative experiments showed that the tritium released could be accounted for almost entirely by authentic [3H]GABA. The electrically evoked overflow of [3H]GABA was tetrodotoxin sensitive and largely calcium-dependent. Exogenous GABA, added to the superfusion medium at 3 to 30 microM, reduced in a concentration-dependent manner the electrically evoked (5 Hz) release of [3H]GABA. The GABAB receptor agonist (-)-baclofen, but not the GABAA receptor agonist muscimol, mimicked GABA and produced a concentration-inhibition curve almost superimposable to that of the natural transmitter. The effects of GABA and of (-)-baclofen were much more pronounced at 5 than at 10 Hz. The GABA-induced inhibition of [3H]GABA release was sensitive to the novel GABAB receptor antagonist beta-(p-chlorophenyl)-3-amino propyl phosphonic acid which, by itself, increased the [3H]GABA overflow. The inhibitory effect of GABA was not counteracted by the GABAA receptor antagonists bicuculline or SR 95531 [2-(3' carbethoxy-2'-propenyl)-3-amino-6-paramethoxy-phenyl-pyr idazinium bromide]. The results are compatible with the presence in the rat cerebral cortex of autoreceptors mediating inhibition of GABA release and belonging to the GABAB type. These autoreceptors may be activated tonically under physiological conditions. PMID- 2547944 TI - Effect of thiorphan on tachykinin-induced potentiation of nerve-mediated contractions of the rat isolated vas deferens. AB - We have assessed the ability of thiorphan, an inhibitor of enkephalinase, to influence the potentiation of the nerve-mediated contractions of the rat isolated vas deferens (pars prostatica) by mammalian tachykinins [substance P, (SP); neurokinin A (NKA); and neurokinin B (NKB)] and selective tachykinin agonists. In the absence of thiorphan, the rank order of potency of mammalian tachykinins was NKA greater than NKB much greater than SP. The maximal response to SP did not exceed 40% of that to NKA or NKB. Thiorphan (10 microM) had no effect on twitches per se, but increased the potency and maximum effect of mammalian tachykinins. [Pro9]-SP sulfone, a selective NK-1 receptor agonist had no effect, either in the absence or presence of thiorphan. [MePhe7]-NKB had some potentiating effect, but only at micromolar concentrations. [Nle10]-NKA (4-10) and [beta-Ala8]-NKA (4-10), two selective NK-2 agonists displayed good activity. [Nle10]-NKA (4-10) was potentiated by thiorphan. On the other hand, the action of [beta Ala8]-NKA (4-10) was completely thiorphan-resistant. These findings indicate that estimate of activity of tachykinins and tachykinin related peptides in this bioassay is influenced markedly by peptide degradation via a thiorphan-sensitive mechanism. NK-2 receptors are the main if not the sole mediators of the response to tachykinins in this bioassay organ. PMID- 2547943 TI - Adrenergic agonists and the Na+-K+-adenosine triphosphatase from rabbit proximal tubules and their basolateral membranes. AB - Several studies suggested that catecholamines modulate renal sodium and water excretion by direct stimulation of adrenergic receptors located on the renal proximal tubule. However, neither the mechanism nor the class of adrenoceptor involved in this effect have yet been established definitively. In the present study, we examined the effects of L-norepinephrine (NE) and selective alpha-1, alpha-2 and beta adrenergic agonists on monovalent cation transport and on Na+-K+ adenosine triphosphatase (ATPase) activity from homogenates, intact tubules and highly purified basolateral membranes prepared from superficial rabbit kidney cortex. Our results showed that neither NE nor specific alpha-1, alpha-2 and beta adrenergic agonists (10 microM) modified ouabain-sensitive uptake of 86Rb+ (a K+ analog) in intact proximal tubules. Similarly, it is demonstrated that NE and alpha and beta adrenergic agonists did not affect Na+-K+-ATPase activity from homogenates, intact tubules and basolateral membranes. The integrity of the alpha 2 adrenergic receptor system, the predominant adrenergic subtype in rabbit proximal tubule, was supported by the following findings: 1) maximal binding of [3H] rauwolscine was about 4-fold higher in basolateral membranes than in homogenates; 2) 5'-guanylimidodiphosphate induced a 27-fold increase in the Ki of NE for alpha-2 receptor in basolateral membranes; 3) NE (5 microM) inhibited by 35% parathyroid hormone-stimulated cyclic AMP production in intact tubules. In conclusion, these data fail to demonstrate that NE, as well as other adrenergic agonists, directly increases Na+-K+-ATPase in the rabbit proximal tubule. Further investigations are needed to clarify the interaction of catecholamines with the renal Na+K+ pump. PMID- 2547945 TI - Changes in cortical acetylcholine and gamma-aminobutyric acid outflow during morphine withdrawal involve alpha-1 and alpha-2 receptors. AB - Naloxone (0.3-9 mumol kg-1), electrical stimulation of locus ceruleus or clonidine at low doses (7.5-112 nmol kg-1) increased the release of acetylcholine from the exposed parietal cortex of freely moving, morphine-tolerant guinea pigs. This increase was not additive and was prevented by prazosin (35.8 nmol kg-1), suggesting the involvement of alpha-1 receptors. At high doses (374 nmol kg-1 or more) clonidine inhibited acetylcholine release through alpha-2 receptors, as it did in naive animals at 7.5 nmol kg-1. Clonidine (374 nmol kg-1) and prazosin (35.8 nmol kg-1) reduced the objective signs of naloxone-precipitated withdrawal. Electrical stimulation of the locus ceruleus or naloxone treatment reduced the release of gamma-aminobutyric acid (GABA) from the exposed parietal cortex of morphine-tolerant guinea pigs. This reduction was not additive and was prevented by idazoxan (84 nmol kg-1), suggesting the involvement of alpha-2 receptors. Clonidine (7.5 nmol kg-1), too, reduced the release of GABA in morphine-tolerant animals. However, when tested jointly with naloxone, clonidine (7.5-112 nmol kg 1) induced alpha-1-mediated facilitation of GABA release (like that elicited in naive animals at 112-374 nmol kg-1) leaving the signs of withdrawal unchanged. This points to the stimulation of alpha-1 receptors highly responsive to this agonist (but not to locus ceruleus stimulation) during naloxone-precipitated withdrawal. In conclusion, chronic morphine treatment modifies the alpha-1- and alpha-2-mediated control of GABA and acetylcholine neurons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547946 TI - Skeletal muscle Na,K-pump concentration in children and its relationship to cardiac glycoside distribution. AB - Skeletal muscle Na,K-pump (cardiac glycoside receptor) concentration was quantified in 18 0- to 8-year-old human subjects by vanadate facilitated [3H]ouabain binding to intact vastus lateralis samples obtained at autopsy. No age-dependent change in [3H]ouabain binding site concentration was observed. Mean value +/- S.E.M. was 268 +/- 17 pmol/g wet wt. (n = 18), range 182 to 433 pmol/g wet wt. At the age of 1 day, 3.5 month and 8 years and 8 months, unspecific uptake and retention of [3H]ouabain was 1.6, 1.4 and 1.5% of the total uptake and retention; release of specifically bound [3H]ouabain during the washout procedure took place with T 1/2 of 97, 90 and 73 hr; and apparent affinity constants for [3H]ouabain binding (KD) was 1.3 x 10(-8), 0.9 x 10(-8) and 1.2 x 10(-8) mol/l. [3H]Ouabain binding site concentrations and kinetics were in agreement with values from adults except that in children apparent affinity constant (KD) was 1.7 times the value in adults. The observation of no age-dependent changes in human skeletal muscle Na,K-adenosine triphosphatase concentration was at variance with the observations of such changes in animals. The total number of Na,K-pumps in the pool of skeletal muscles increased from 10 to 50 times that in the heart from birth to old age. The skeletal muscle pool of Na,K-pumps seems to constitute a distribution volume of importance during digitalization in children as well as adults.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2547947 TI - Detection of human papillomavirus (HPV) DNA in focal epithelial hyperplasia. AB - Five focal epithelial hyperplasia (FEH) specimens from four patients were examined by Southern blot hybridization analysis to determine the specific human papillomavirus (HPV) types present. The histomorphologic features of these specimens were also evaluated and a broad variety of changes including koilocytes, mitosoid cells, ballooning cells and cells showing individual cell keratinization were noted. FEH lesions from the three patients sharing a familial relationship demonstrated HPV DNA sequences that were either the prototype HPV-13 or a very closely related HPV-13 subtype. These patients also showed similar clinical features. Lesional tissue from the other patient was found to harbor HPV DNA sequences similar to HPV-32. In view of these findings it is suggested that these specific HPV types are associated with the characteristic FEH histomorphology described. PMID- 2547948 TI - Beta-adrenergic receptor concentration and subtype in the corpus luteum of the adult pseudopregnant rat. AB - Luteal beta-adrenergic receptor concentration and subtype were determined in adult pseudopregnant rats during and after the period of the functional luteal phase. The specific beta-adrenergic receptor ligand (-)-3-[125I]iodocyanopindolol ([125I]ICYP) was used to determine the receptor concentration in corpora lutea of adult pseudopregnant rats. A 3-fold increase in beta-adrenergic receptor concentration was seen during the first 2-3 days of pseudopregnancy, whereafter the receptor concentration declined. During the functional luteal regression period (Day 12-15) the receptor levels were still low. In regressed (Day 16-22) corpora lutea a temporary increase in beta-receptor concentration was seen which may represent some role for beta-adrenergic mechanisms in the regulation of morphological regression in the corpus luteum. To determine the beta-adrenergic subtype, competition of [125I]ICYP-binding with selective beta 1- and beta 2 adrenergic antagonists was assessed in corpora lutea of different ages and in rat heart and uterus. The beta-adrenergic receptors in corpora lutea of adult pseudopregnant rats were shown to be solely of the subtype beta 2, regardless of the luteal age. PMID- 2547949 TI - Guinea pig testicular proacrosin-acrosin system: preliminary immunological characterization. AB - In this study, immunochemical techniques were employed to examine the guinea pig (GP) testicular proacrosin-acrosin system. Monospecific polyclonal antibodies to the highly stable enzymatically active 34,000 molecular weight form of GP testicular acrosin were generated. Western blot analysis of acid extracts prepared from snap-frozen freshly excised GP testes revealed two major immunoreactive bands with mol. wts of approximately 62,000 and 48,000 and one minor band with an approximate mol. wt of 54-56,000. The 62,000 mol. wt molecule identified is in close agreement with the previously reported mol. wt for purified GP testicular proacrosin. Western blot analysis of different species of testicular acid extracts demonstrated the evolutionary relatedness of the proacrosin-acrosin systems since immunoreactivity was observed primarily with acid extracts from rodent species (rat, mouse and hamster) and not with extracts from evolutionarily less-related species (goat, ram and bovine). The majority of the cross-reactivity observed was characterized by immunoreactivity with the 62,000 and 48,000 mol wt molecules. The only species which exhibited cross reactivity with the 54-56,000 mol. wt protein was rat. In addition, the iso immunogenic and aspermatogenic properties of the 34,000 mol. wt form of GP testicular acrosin were examined. One out of five Hartley and one out of seven Strain 2 female GPs immunized and boosted with a total of 200 micrograms of purified protein exhibited increased levels of circulating anti-acrosin iso antibodies. The antigenic specificity of the iso-antibodies observed in the two animals was verified by Western blot analysis. All other female animals, including two strain 13 GPs, exhibited very low or undetectable levels of such antibodies following immunization. One out of three male Hartley GPs immunized with 50 micrograms of the purified protein exhibited typical lesions of experimental allergic orchitis while none of a group of three animals developed lesions at a 5 micrograms dose. Taken together, these results suggest that the 34,000 mol. wt form of GP testicular acrosin is neither a highly potent iso immunogen nor aspermatogenic autoantigen. PMID- 2547951 TI - An association between mineral metabolism and the renin-aldosterone system in human hypertension. AB - In recent years abnormalities of calcium metabolism have been described in human hypertension. In this study, the relationships between indices of calcium metabolism and the renin-aldosterone system were studied in 39 subjects with untreated essential hypertension. No significant associations were found between the major determinants of calcium metabolism (plasma ionised calcium, parathyroid hormone and 1,25(OH)2-vitamin D) and the renin-aldosterone system. Serum magnesium was, however, positively correlated to plasma renin activity (PRA) (r = 0.38, P less than 0.05) while both 24 h urinary excretion of calcium and cAMP were found to be correlated to both PRA and urinary aldosterone in a positive way (r = 0.39-0.42, P less than 0.01 and r = 0.33-0.57, P less than 0.01, respectively). In this study there was no other evidence of any major influence of the renin-aldosterone status on the calcium balance in human hypertension. The urinary leak of calcium might be determined by the action of the renin aldosterone system. PMID- 2547950 TI - A randomised double blind comparison of enalapril versus hydrochlorothiazide in elderly hypertensives. AB - To assess the suitability of therapy with an angiotensin-converting enzyme inhibitor for older hypertensives, a randomised double blind two group comparison of enalapril (10 to 40 mg daily) with hydrochlorothiazide (25 to 100 mg daily) was carried out in 32 untreated and independently living subjects aged 63 to 82 years with pressures on screening greater than 100 mmHg diastolic or greater than 180 mmHg systolic and after two weeks on placebo greater than 90 mmHg diastolic or greater than 160 mmHg systolic. Twenty-five subjects completed the ten weeks of active treatment with the majority in both groups reaching target blood pressures. Four subjects on enalapril and three on hydrochlorothiazide withdrew because of side effects. Eighteen of the 32 patients volunteered troublesome symptoms they attributed to therapy, which in some instances were alleviated by reductions in drug dosage. Small but significant falls in plasma potassium and sodium and increases in plasma urea and uric acid were seen on hydrochlorothiazide only. It was concluded that although in most cases good blood pressure control can be achieved by enalapril with some advantages over other drugs in the elderly, the potential for adverse effects remains significant. Halving usual starting doses of this and other antihypertensives may help to minimise problems in older subjects. PMID- 2547952 TI - Cardiovascular, renal and endocrine responses to low doses of atrial natriuretic factor in mild essential hypertension. AB - The purpose of this study was to evaluate the cardiovascular, renal and endocrine effects of human atrial natriuretic factor (ANF), infused at a rate of 0.8 microgram/min (about 4 pmol/kg/min) for three hours in normal subjects and patients with essential hypertension. This infusion rate was chosen to obtain a range of plasma ANF levels which can be generated by physiological manoeuvres and to reduce the likelihood of hypotension. Five patients and six healthy volunteers participated in the study. The infusion had to be prematurely discontinued in one patient and in one control because of hypotension with relative bradycardia. Blood pressure otherwise remained unchanged during infusion whereas heart rate rose transiently. Plasma ANF levels increased similarly during infusion from 8.9 +/- 2.6 to 23.9 +/- 6.4 pmol/l in patients and from 3.7 +/- 0.7 to 25.4 +/- 6.9 pmol/l in the controls, remained stable during the infusion, and decreased similarly in both groups after the infusion, with a half-life of 7 min. Plasma guanosine cyclic phosphate (cGMP) was augmented by about four-fold in both groups. In both groups, plasma aldosterone levels fell whereas plasma noradrenaline increased. The diuretic effect of ANF was similar in both controls and patients (1354 +/- 161 vs 1542 +/- 116 ml/3 hrs respectively), whereas its natriuretic effect was exaggerated in hypertensive patients (90 +/- 11 vs 62 +/- 9 mmol/3 hrs, P less than 0.05). In conclusion, this low infusion rate of ANF produced similar changes in plasma ANF, cGMP, aldosterone and noradrenaline levels but patients with mild essential hypertension demonstrated an exaggerated diuretic and natriuretic response to ANF infusion. PMID- 2547954 TI - Guidelines for the prevention of simian immunodeficiency virus infection in laboratory workers and animal handlers. AB - The authors are members of a working group that formulated guidelines to minimize transmission of simian immunodeficiency virus (SIV) infection to man. Biosafety level (BSL) 2 standards are recommended for handling of clinical specimens and housing of SIV inoculated animals. Manipulation of SIV preparations may be performed in a BSL 2 facility with additional BSL 3 practices and equipment; for large volume or concentrated preparations of SIV BSL 3 containment is necessary. Written policies regarding management and testing of workers exposed to SIV are recommended. PMID- 2547953 TI - Serum and tissue positivity for hepatitis B virus markers in histopathologically proven glomerulonephropathies. AB - To assess the pathogenic significance of hepatitis B virus (HBV) in glomerulonephritis (GN), 98 patients with histopathologically proven glomerulonephropathies were screened for HBV markers, complement components and levels of circulating immune complexes (CICs); and renal biopsies from 31 of them were examined for the presence of hepatitis B surface antigen (HBsAg), and its location, by immunoperoxidase staining. The HBsAg positive rate in the patients (who came from a population with 10% HBsAg positivity) ranged from 51.9% in minimum change nephrotic syndrome (MCNS) to 81.8% in patients with proliferative glomerulonephritis (PGN). Whereas 24.5% of the cases were positive for HBsAg only, 10.2% had anti-HBcIgM with HBsAg, 13.3% had HBeAg with HBsAg and 9.2% had HBsAg, HBeAg and anti-HBcIgM. Complement component C3 levels were decreased in all groups of GN studied, but C4 levels varied. CIC levels were significantly increased (p less than 0.01) only in HBsAg-positive MCNS, focal glomerulosclerosis (FGS) and membranous glomerulonephritis (MGN). Of the 31 renal biopsies examined for the deposition of HBsAg, 4 (12.9%) were found to be positive for HBsAg in situ; 64.5% of biopsied patients were seropositive for HBsAg and 77.4% had CICs. All the four in-situ HBsAg-positive cases were seropositive for HBsAg, HBeAg and anti-HBcIgM with significantly high CIC levels (p less than 0.01). HBsAg deposition was intracytoplasmic in the mesangial cells of the glomeruli, in the glomerular basement membrane or in the tubules, or in a combination of these sites. PMID- 2547955 TI - Monoclonal antibodies to type D retrovirus (SRV-2). AB - Monoclonal antibodies (MoAbs) to the major gag core protein p27 and a viral protein p44 of type D retrovirus (SRV-2) were produced and used in the detection of SRV-2 antigens in infected Raji cells and in tissues from macaques with simian acquired immunodeficiency syndrome (SAIDS) and retroperitoneal fibromatosis (RF). Anti-p44 MoAb showed inhibition of syncytium formation by both SRV-1- and SRV-2 infected Raji cells. PMID- 2547956 TI - Serologic and virologic analysis of type D simian retrovirus infection in a colony of Celebes black macaques (Macaca nigra). AB - Celebes macaques were tested for type D simian retrovirus (SRV) infection. SRV infection was first detected in one serum sample collected during 1980. By 1983, 32 of 46 monkeys (70%) were infected. Serotyping of the SRV isolates determined that 0/26 of the isolates were SRV-1; 24/26 were SRV-2; 1/26 was SRV-5; and 1/26 could not be typed. Restriction endonuclease mapping confirmed the SRV-2C and SRV 5 isolates. In addition, two SRV-2C variants were detected. PMID- 2547957 TI - Mucosal epithelial cells and Langerhans cells are targets for infection by the immunosuppressive type D retrovirus simian AIDS retrovirus serotype 1. AB - Simian acquired immune deficiency syndrome (SAIDS) caused by the type D retrovirus SRV-1 results in opportunistic infections and a spectrum of oral lesions similar to those seen in humans with AIDS. To better understand the pathogenesis of these oral lesions we have retrospectively examined the oral mucosa from ten rhesus monkeys that died with SAIDS and prospectively examined the oral mucosa of ten additional animals inoculated with SRV-1 to determine at what time, and in what cells SRV-1 infection of the oral mucosa occurs. Using single and double label immunohistologic techniques, and electron microscopy we detected SRV-1 in clusters of oral epithelial cells and rare Langerhans cells as early as 1 month postinoculation. PMID- 2547958 TI - Detection of simian immunodeficiency virus in macaque lymph nodes with a SIVmac envelope probe. AB - A tritium-labeled DNA envelope gene probe was used to detect Simian Immunodeficiency Virus in formalin fixed lymph nodes from rhesus monkeys experimentally inoculated with SIVmac251. Cells containing SIV RNA produced strong hybridization signal and were present in small numbers in biopsy specimens and in much greater numbers in lymph nodes collected at autopsy. SIV-infected cells were morphologically identified as lymphocytes and macrophages. PMID- 2547959 TI - Transmission of the simian immunodeficiency virus SIVmne in macaques and baboons. AB - A primate lymphotropic lentivirus was isolated on Hut 78 cells after cocultivation of a lymph node from a macaque that died with malignant lymphoma. In earlier studies SIV/Mne was inoculated into 17 macaques and two baboons. All of the macaques became viremic and seropositive. Fifteen of the macaques succumbed to a classic AIDS-like disease, whereas the baboons did not become viremic. The SIV/Mne virus has now been molecularly cloned and inoculated into Macaca nemestrina and baboons. A new transmission study has been initiated to test the effects of route and dosage on disease. PMID- 2547961 TI - Genetic analysis and infection of SIVAGM and SIVMND. AB - Recently, the authors determined the partial sequence of simian immunodeficiency virus (SIV) from the mandrill (SIVMND) and found SIVMND to be a new member of the HIV/SIV group, equidistant from other members, including SIVAGM. Experimentally, the African green monkey and cynomolgus monkey could be infected with SIVAGM and the cottontop tamarin with SIVMND. However, no clinical sign of an AIDS-like disease was observed in these monkeys. PMID- 2547960 TI - Infection of macaque monkeys with simian immunodeficiency virus from African green monkeys: virulence and activation of latent infection. AB - The virulence of three isolates of simian immunodeficiency virus from African green monkeys (SIVagm) was studied in rhesus and pigtailed macaques. None of 15 rhesus monkeys and one of four pigtailed monkeys died from infection during the time they were studied (up to 33 months). SIVagm was only isolated from rhesus monkeys for up to 2 months after inoculation. However, when these animals were secondarily infected with Simian acquired immunodeficiency syndrome retrovirus type 1 (SRV-1), SIVagm was activated and isolated. Dual infection caused increased mortality. PMID- 2547962 TI - Genetic diversity among simian immunodeficiency virus isolates from African green monkeys. AB - To characterize isolates further within the SIVagm subtype, we studied four SIVagm isolates by cross-hybridization, molecular cloning, and nucleotide sequencing. Our results indicate an unexpected degree of genetic variation among isolates within the SIVagm subtype comparable to the variation between SIVmac and HIV-2. PMID- 2547963 TI - Molecular cloning of SIV from sooty mangabey monkeys. AB - We derived two infectious molecular clones of SIV from sooty mangabey monkeys (Cercocebus atys) and compared them by restriction enzyme mapping and limited DNA sequencing to other known primate lentiviruses. These analyses show that SIVsmm is closely related to, but distinct from, SIVmac and HIV-2. Our data suggest that SIVmac may have been derived from SIVsmm by cross-species transmission in captivity. PMID- 2547964 TI - Molecular characterization and comparison of simian immunodeficiency virus isolates from macaques, mangabeys, and African green monkeys. AB - Simian immunodeficiency virus (SIV)/Mne has been inoculated into three species of macaques and into baboons. Virus was isolated from all the macaques who subsequently died at 15 to 120 weeks (mean 80 weeks) with various manifestations of immune deficiency. Individual animals varied in their viral antibody profile as a function of time after infection. Independent SIV isolates obtained from African green monkeys and magabeys were compared to SIV/Mne for their ability to replicate in lymphocytes and macrophages and with respect to the immunological relatedness of their viral proteins. Antibodies present in human immunodeficiency virus-2 (HIV-2)-infected individuals were readily detected by the virus produced by a single-cell clone of SIV/Mne. PMID- 2547965 TI - Rhesus macaques inoculated with molecularly cloned simian immunodeficiency virus. AB - We have isolated a biologically active molecular clone of simian immunodeficiency virus (SIV), SIVmac 1A11, originally obtained from a rhesus macaque at the New England Regional Primate Research Center. Virus derived from cells transfected with this clone is cytopathic for rhesus peripheral blood mononuclear cells, replicates in cultures of rhesus macrophages, and infects rhesus macaques when inoculated intravenously. Six macaques inoculated with SIVmac 1A11 all became infected and produced antibodies to viral envelope glycoproteins that neutralized virus. Antibodies to viral core proteins were detected in only one animal. No clinical signs of disease were observed throughout 7 months postinoculation. PMID- 2547966 TI - Postexposure immunotherapy of simian immunodeficiency virus (SIV) infected rhesus with an SIV immunogen. AB - An inactivated whole simian immunodeficiency virus (SIV) immunogen given to healthy, seropositive rhesus macaques 4 months after infection had no effect on the humoral immune response to SIV, the presence of antigenemia, cell-associated viremia, or disease course. Further immunotherapeutic trials in this highly susceptible animal model should be carried out sooner after exposure, before significant loss of CD4 cells has occurred. The SIV infected macaque model will continue to serve an essential role in development and testing of anti-AIDS drugs and immunogens. PMID- 2547967 TI - Proliferative T-cell response to HIV envelope glycoprotein in immunized and infected primates and human beings. AB - Human immunodeficiency virus (HIV)-specific helper T-cell response was studied in human subjects and nonhuman primates either infected with HIV or immunized with different HIV protein preparations. A strong group-specific T-cell response involving T-cell proliferation and lymphokine secretion was observed in immunized chimpanzees and rhesus monkeys as well as HIV-infected chimpanzees and gibbons. HIV-infected people demonstrated a low or no HIV-specific T-cell response. In contrast, five of 14 HIV antibody-negative sexual partners of HIV-infected men recognized one or more T-cell epitopes in the envelope glycoprotein of HIV. PMID- 2547968 TI - Tetracycline promoter mutations decrease non-B DNA structural transitions, negative linking differences and deletions in recombinant plasmids in Escherichia coli. AB - The ability to clone a variety of sequences with varying capabilities of adopting non-B structures (left-handed Z-DNA, cruciforms or triplexes) into three loci of pBR322 was investigated. In general, the inserts were stable (non-deleted) in the EcoRI site (an untranslated region) of pBR322. However, sequences most likely to adopt left-handed Z-DNA or triplexes in vivo suffered deletions when cloned into the BamHI site, which is located in the tetracycline resistance structural gene (tet). Conversely, when the promoter for the tet gene was altered by filling-in the unique HindIII or ClaI sites, the inserts in the BamHI site were not deleted. Concomitantly, the negative linking differences of the plasmids were reduced. Also, inserts with a high potential to adopt Z-DNA conformations were substantially deleted in the PvuII site of pBR322 (near the replication origin and the copy number control region), but were less deleted if the tet promoter was insertion-mutated. The deletion phenomena are due to the capacity of these sequences to adopt left-handed Z-DNA or triplexes in vivo since shorter inserts, less prone to form non-B DNA structures, or random sequences, did not exhibit this behavior. Sequences with the potential to adopt cruciforms were stable in all sites under all conditions. These results reveal a complex interrelationship between insert deletions (apparently the result of genetic recombination), negative supercoiling, and the formation of non-B DNA structures in living Escherichia coli cells. PMID- 2547969 TI - Crystallization and preliminary X-ray studies of Escherichia coli glycerol kinase. AB - Escherichia coli glycerol kinase, a major regulatory enzyme which catalyzes the reversible MgATP-dependent phosphorylation of glycerol has been crystallized by the hanging drop vapor diffusion method at room temperature. Three different crystal forms have been obtained in the presence of glycerol and appear to be suitable for X-ray crystallographic studies. Vapor diffusion against 55% ammonium sulfate and 1% beta-octyl glucoside (pH 7.0) yields rhombohedral crystals with space group R32, a = b = 277.1 A, c = 78.7 A (hexagonal indexing) containing a dimer of Mr 112,000 in the asymmetric unit (Vm = 2.64 A3/dalton). Vapor diffusion against sodium chloride in the presence of 10% (w/v) polyethylene glycol (pH 6.5 to 7.0) yields two different crystal forms, both with space group P2(1). The first form has a = 88.1 A, b = 99.3 A, c = 114.6 A, beta = 119 degrees, the second form has a = 92.5 A, b = 117.6 A, c = 108.3 A, beta = 93.64 degrees. Addition of ADP enhances growth of the monoclinic forms. These forms appear to contain an entire tetramer of Mr 224,000 in the asymmetric unit and have Vm values of 2.28 and 2.65 A3/dalton, respectively. All forms diffract to better than 3.0 A resolution while the second monoclinic form diffracts to approximately 1.8 A. PMID- 2547970 TI - Activation of the lac promoter and its variants. Synergistic effects of catabolite activator protein and supercoiling in vitro. AB - Escherichia coli lac promoter variants are shown to be subject to large synergistic transcriptional activation by catabolite activator protein (CRP) and DNA supercoiling in vitro. Activation was studied for the lac wild-type promoter, a promoter with a variant spacing (lac delta l) and two promoters with variant 10 regions (lac ps, lac UV5). The variant promoters respond to the simultaneous presence of CRP and supercoiling by exhibiting large multiplicative activation at the low to moderate superhelicities that are most pertinent in vivo. Although all four promoters can be activated by CRP, those made stronger by changing downstream promoter elements are less CRP-activated even though each contains an identical CRP binding site. When each of the variant promoters is made stronger by introducing DNA supercoils, the apparent CRP activation initially remains constant but eventually declines at higher superhelicities. Thus, strengthening the lac promoter through either DNA sequence changes or the introduction of high level DNA supercoiling can lead to diminished potential for activation by CRP. These results are interpreted in terms of a role for CRP in providing extra stabilizing contacts for RNA polymerase binding that are necessary only when other stabilizing features of promoter structure are lacking. PMID- 2547971 TI - Determinants of site-specific recombination in the lambdoid coliphage HK022. An evolutionary change in specificity. AB - The temperate bacteriophage HK022, like its relative lambda, inserts its chromosome into a specific site in the bacterial chromosome during lysogenization and excises it after induction. However, we find that the recombinational specificities of the two phages differ: they use different bacterial sites, and neither promotes efficient insertion or excision of the other phage chromosome. In order to determine the basis for this difference in specificity, we sequenced the HK022 elements that are involved in insertion and excision, and compared them to the corresponding lambda elements. The location, orientation, size and overall arrangement of the int and xis genes and the phage attachment sites are nearly identical in the two genomes, as is common for other functionally related elements in lambdoid phages. The Xis proteins of the two phages are functionally interchangeable, and their predicted amino acid sequences differ by but one residue. In contrast, the two Int proteins are not functionally interchangeable, and their sequences, although similar, differ at many positions. These sequence differences are not uniformly distributed: the amino-terminal 55 residues are completely conserved, but the remaining 302 show a pattern of differences interspersed with identities and conservative changes. These findings imply that the specificity difference between HK022 and lambda site-specific recombination is a consequence of the inability of the respective Int proteins to recognize pairs of heterologous attachment sites. The two phage attachment sites are remarkably similar, especially the two "arm" segments, which in lambda contain binding sites for Int, Xis and integration host factor. They are less similar in the segment between the two arms, which in lambda contains the points of recombinational strand exchange and a second class of binding site for Int protein (the "core-type" sites). The two bacterial attachment sites are quite different, although both have a short stretch of perfect homology with their respective phage partners at the points of strand exchange. We propose that the two Int proteins recognize similar or identical sites in the arms of their cognate attachment sites, and that differences in binding or action at the core type sites is responsible for the divergent specificities. Genetic experiments and sequence comparisons suggest that both proteins recognize different but overlapping families of core-type sites, and that divergence in specificity has been achieved by an alternating succession of small, mutually compatible changes in protein and site. PMID- 2547973 TI - Receptor specificity of the Escherichia coli T-even type phage Ox2. Mutational alterations in host range mutants. AB - The T-even type Escherichia coli phage Ox2 uses the outer membrane protein OmpA as a receptor. The protein is recognized with the ends of the virion's long tail fibers. The 266 residue protein 38 is located at this site and acts as an adhesin. Host-range mutants had previously been isolated from Ox2. Mutant Ox2h5 is able to infect cells possessing an altered OmpA protein, which renders the cell resistant to Ox2. Ox2h10 was selected from Ox2h5. This phage recognizes the OmpC protein in addition to the OmpA protein. Ox2h12, which stems from Ox2h10, binds to OmpC with high affinity, but has lost efficient binding to OmpA. The mutational alterations caused in genes 38 are: Asp231----Asn(h5) and His170--- Arg(h10). The triple mutant Ox2h12 possesses an insertion of a Gly residue next to Gly121. The three mutants have additionally acquired mutations affecting their base plate, making them "trigger-happy". When protein 38 was compared with the same protein derived from other E. coli phages, it was found to contain two constant and one variable domains, the latter harboring four hypervariable regions flanked by a largely conserved glycine-rich sequence. The h5 and h10 mutations occurred within two hypervariable areas, while the additional Gly residue was present in one of the flanking conserved sequences. On the basis of these results, as well as those obtained from host-range mutants analyzed previously, a model for such adhesins is proposed. Receptor recognition is most likely performed via the hypervariable regions, which may form loops held together in close proximity by the oligoglycine sequences. The latter may achieve this by being part of highly compact omega loops. PMID- 2547972 TI - Ligand-modulated binding of a gene regulatory protein to DNA. Quantitative analysis of cyclic-AMP induced binding of CRP from Escherichia coli to non specific and specific DNA targets. AB - This paper describes a generally applicable method for quantitative investigation of ligand-dependent binding of a regulatory protein to its target DNA at equilibrium. It is used here to analyse the coupled binding equilibria of cAMP receptor protein from Escherichia coli K12 (CRP) with DNA and the physiological effector cAMP. In principle, the DNA binding parameters of CRP dimers with either one or two ligands bound are determinable in such an approach. The change of protein fluorescence was used to measure CRP binding to its recognition sequence in the lac control region and to non-specific DNA. Furthermore, the binding of cAMP to preformed CRP-DNA complexes was independently studied by equilibrium dialysis. The data were analysed using a simple interactive model for two intrinsically identical sites and site-site interactions. The intrinsic binding constant K and the co-operativity factor alpha for binding of cAMP to free CRP depend only slightly on salt concentration between 0.01 M and 0.2 M. In contrast, the affinity of cAMP for CRP pre-bound to non-specific DNA increases with the salt concentration and the co-operativity changes from positive to negative. This results from cation rebinding to the DNA lattice upon forming the cAMP-CRP-DNA complex from cAMP and the pre-formed CRP-DNA complex. The CRP-cAMP1 complex shows almost the same affinity for specific and non-specific DNA as the CRP-cAMP2 complex, and both displace the same number of cations. It is concluded that the allosteric activation of CRP is induced upon binding of the first cAMP. These results are used to estimate the occupation of the CRP site in the lac control region in relation to the cAMP concentration in vivo. Under physiological conditions the lac promoter is activated by the CRP dimer complexed with only one cAMP. Furthermore, a model for the differential activation of various genes expressed under catabolite repression is presented and discussed. PMID- 2547974 TI - Crystallization of the DpnM methylase from the DpnII restriction system of Streptococcus pneumoniae. AB - Three proteins, two DNA methylases and an endonuclease, from the DpnII restriction system of Streptococcus pneumoniae recognize the DNA sequence 5' GATC 3' but have very different amino acid sequences, which make them interesting subjects for structural determination. A purification procedure was developed that conveniently yields milligram amounts of the DpnM methylase. The DpnM protein tends to precipitate at reduced ionic strength, and this property was exploited to yield well-formed bipyramidal crystals. By X-ray diffraction, the crystals of DpnM were found to be orthorhombic, with cell dimensions a = 56.9 A, b = 68.2 A, c = 84.5 A; systematic absences identify the space group as P2(1)2(1)2(1). Diffraction extends beyond 3 A, so the crystals may allow structural determination at atomic resolution. PMID- 2547975 TI - Evidence that mutation patterns vary among Drosophila transposable elements. AB - In Drosophila melanogaster, codon usage in the open reading frames (ORFs) of transposable elements (TEs) differs greatly from that in other ORFs. In addition, while the ORFs from a single element are similar, there is considerable variation among elements. In the TE ORFs there are no indications of selection for the codons prevalent in the other D. melanogaster genes, but rather codon usage can be succinctly summarized in terms of the base composition at silent sites. We suggest that the particular silent site base composition of each TE is determined by an individual pattern of mutation. In many of the TEs there is an ORF encoding a protein with homology to reverse transcriptase; the amino acid sequences of these are quite divergent, and so it is possible that each of these incorporates certain mismatched bases at different frequencies during replication. PMID- 2547976 TI - Crystallization of a complex between ribonuclease Ms and 3'-guanylic acid. AB - The crystals of a complex between ribonuclease Ms, the extracellular ribonuclease from Aspergillus saitoi, and 3'-guanylic acid were obtained from 2-methyl-2,4 pentanediol solution by vapor diffusion technique in the hanging drop mode. The crystals belong to orthorhombic space group P2(1)2(1)2(1) with dimensions a = 47.0 A, b = 62.8 A, c = 37.9 A. The crystals diffract strongly up to at least 2.0 A resolution. PMID- 2547977 TI - Mobilization of lead by esters of meso-2,3-dimercaptosuccinic acid. AB - An examination was made of the relative efficacies of 2,3-dimercapto-1-propanol (BAL) and three diesters ( [CH(SH)COOR]2; DMDMS, R = CH3; DEDMS, R = C2H5; and Di PDMS, R = CH(CH3)2] of meso-2,3-dimercaptosuccinic acid (DMSA) in mobilizing freshly injected lead from mice. These diesters, like BAL, reduced the lead levels resulting from freshly injected lead in both the soft tissues (liver, kidneys, spleen, and brain) and the bone (tibia). After treatment with the dimethyl (DMDMS), the diethyl (DEDMS), and the diisopropyl (Di-PDMS) esters the lead content of each of the organs was significantly less than that present in the untreated controls. Each of the diesters reduced lead levels in the kidneys, liver, and spleen significantly below those levels found after BAL treatment. The action of the diesters in reducing brain lead levels was comparable to that of BAL. Di-PDMS was the most effective of these compounds and was significantly superior to BAL. Each of the esters was also significantly more effective than BAL in reducing the whole body level of lead. PMID- 2547979 TI - Developmental and regional expression of three new members of the ras-gene family in the mouse brain. AB - We have examined the expression in the mouse nervous system of three new members of the ras protooncogene family: rab1, rab2, and rab3. Each of these genes was transcribed into messenger RNAs with different molecular weights. These transcripts has specific developmental and regional patterns of expression. In particular, for the three genes, the ratio between the heavy and light mRNAs depended strongly on developmental stage and brain region. The use of pure neuronal and glial cultures revealed that the high molecular weight transcripts were enriched in neurons and that, in the case of rab2 and rab3, their expression increased with neuronal differentiation. These results are discussed considering the sequence identities between these genes and the yeast YPTI and sec-4 genes, which are known to be implicated in post-Golgi vesicular transport and cytoskeletal stabilization. We propose that the rab genes might be of importance in the regulation of these two processes within the developing and adult nervous system. PMID- 2547978 TI - The chemical properties of silica particle surface in relation to silica-cell interactions. AB - Although silicosis has been studied extensively, the mechanism is still not fully understood. Experiments do provide evidence that the actions of unique properties of silica surface on the cell membrane are the starting point of silicotic processes. This paper summarizes literature on chemical properties of silica surface, and the effect of particle size on silica toxicity. This paper also discusses the ways in which silica dusts are thought to interact with the cell membrane, with emphasis on freshness, hydrogen bonding, and free-radical interactions. PMID- 2547980 TI - Characterization of beta-adrenoceptors in the Syrian hamster Harderian gland: sexual differences and effects of either castration or superior cervical ganglionectomy. AB - Scatchard analysis of saturation isotherms of [125I]-iodopindolol was used to characterize beta-adrenoceptor density (Bmax) and affinity constant (Kd) in female and male hamster Harderian glands. Single-point experiments were also completed in intact females, intact males, and castrated or superior cervical ganglionectomized males. Scatchard analysis described a single population of binding sites with a Bmax of 292.2 +/- 45.1 fmol/mg protein (X +/- SEM, n = 6) in females and 18.2 +/- 3.0 fmol/mg protein (n = 6, P less than .001) in males. The affinity also varied significantly (P less than .05) with a Kd of 1.08 +/- 0.18 versus 0.26 + 0.05 nM (n = 6) in the Harderian gland of females and males, respectively. Single-point [125I]-IPIN (400 pM) binding values in females were 67.3 +/- 4.0, in intact males were 12.8 +/- 3.2, and in castrated males were 31.2 +/- 4.2 fmol/mg protein (n = 7-9). Superior cervical ganglionectomy induced no significant changes in receptor binding. The results indicate pronounced sexual differences in the density and affinity of beta-adrenoceptors in the hamster Harderian gland, which may be sex hormone dependent. PMID- 2547981 TI - Immunohistochemical research on HDV infection in Chinese patients with chronic liver disease. AB - The prevalence of HDAg in the liver of Chinese patients with chronic hepatitis and hepatocellular carcinoma was determined by using direct immunofluorescence and immunoperoxidase. Six patients (6.3%) out of 95 HBsAg carriers with inflammatory liver disease and neoplasia were found to be HDAg positive. HDAg was shown in the liver of 6 patients (7.6%) among 79 patients with chronic hepatitis. The relative frequency of HDAg in cirrhosis-B, chronic active hepatitis B and chronic persistent hepatitis B was 14.3%, 7.1% and 5.9% respectively. These results suggest that a sizeable number of HBsAg carriers are superinfected with HDV. In view of the large amount of HBV carriers in China, the relative minor but distinct presence of HDV poses an important community health problem. PMID- 2547982 TI - Haemoglobin genotypes and primary liver cell carcinoma in Africans. AB - The distribution of haemoglobin genotype in 200 patients with primary liver cell carcinoma (PLCC) did not show any statistical difference from those of 200 age and sex-matched controls. It is therefore concluded that haemoglobin genotype is unlikely to play a significant role in the development of PLCC in Africans. PMID- 2547983 TI - Prevalence of antibody to hepatitis A virus in the Santa Cruz region of Bolivia. AB - Several epidemiological studies have shown the worldwide distribution of hepatitis A with the highest prevalence of anti-HAV antibodies in developing countries (Papaevangelou 1984). There is no information about the epidemiology of hepatitis A in Bolivia. The goal of this study was to evaluate the anti-HAV antibody prevalence in the Santa Cruz region, southeastern Bolivia. PMID- 2547984 TI - Sonography of abdominal desmoid tumors in adolescents. PMID- 2547985 TI - Leukemogenic membrane glycoprotein encoded by Friend spleen focus-forming virus: transport to cell surfaces and shedding are controlled by disulfide-bonded dimerization and by cleavage of a hydrophobic membrane anchor. AB - The leukemogenic glycoprotein (gp55) encoded by Friend spleen focus-forming virus is predominantly retained in the rough endoplasmic reticulum (RER). However, a small proportion (ca. 5%) is processed to form a derivative that occurs on plasma membranes and causes mitosis of infected erythroblasts. We have now found that gp55 folds heterogeneously in the RER to form components with different disulfide bonds and that this difference may determine their processing fates. RER gp55 consists predominantly of monomers with intrachain disulfide bonds. In contrast, the processed molecules are disulfide-bonded dimers. These dimers are extensively modified in transit to cell surfaces by conversion of four N-linked high-mannose oligosaccharides to complex derivatives and by attachment of a sialylated O linked oligosaccharide. The plasma membrane dimers are then slowly shed into the medium by a mechanism that involves proteolytic cleavage of approximately 25 membrane-anchoring hydrophobic amino acids from the carboxyl termini of the glycoproteins. Consequently, shed molecules have shorter polypeptide chains than cell-associated gp55. We conclude that gp55 folds into different disulfide-bonded components that do not substantially isomerize, and that only one specific dimer is competent for export from the RER. Mitogenic activity of gp55 could be caused by the cell surface dimers, by the shed derivative, or by the carboxyl-terminal hydrophobic anchors that remain in the membranes after the shedding reaction. PMID- 2547986 TI - Glycoprotein cytoplasmic domain sequences required for rescue of a vesicular stomatitis virus glycoprotein mutant. AB - We have used transient expression of the wild-type vesicular stomatitis virus (VSV) glycoprotein (G protein) from cloned cDNA to rescue a temperature-sensitive G protein mutant of VSV in cells at the nonpermissive temperature. Using cDNAs encoding G proteins with deletions in the normal 29-amino-acid cytoplasmic domain, we determined that the presence of either the membrane-proximal 9 amino acids or the membrane-distal 12 amino acids was sufficient for rescue of the temperature-sensitive mutant. G proteins with cytoplasmic domains derived from other cellular or viral G proteins did not rescue the mutant, nor did G proteins with one or three amino acids of the normal cytoplasmic domain. Rescue correlated directly with the ability of the G proteins to be incorporated into virus particles. This was shown by analysis of radiolabeled particles separated on sucrose gradients as well as by electron microscopy of rescued virus after immunogold labeling. Quantitation of surface expression showed that all of the mutated G proteins were expressed less efficiently on the cell surface than was wild-type G protein. However, we were able to correct for differences in rescue efficiency resulting from differences in the level of surface expression by reducing wild-type G protein expression to levels equivalent to those observed for the mutated G proteins. Our results provide evidence that at least a portion of the cytoplasmic domain is required for efficient assembly of the VSV G protein into virions during virus budding. PMID- 2547987 TI - Functional interaction of constant and variable domains of human immunodeficiency virus type 1 gp120. AB - A previously reported amino acid substitution within the second conserved domain of the human immunodeficiency virus type 1 (HIV-1) gp120 envelope results in the production of noninfectious particles. Molecular characterization of spontaneous revertant viruses, which arose during long-term cocultures of this env mutant, revealed that an amino acid change within another region of gp120 could functionally compensate for the mutation and restore infectivity. In the current study, we have introduced a conservative amino acid substitution at this second site revertant codon and observed a marked reduction in HIV-1 infectivity. During the passage of this defective virus in cocultures, yet another revertant appeared which contained an amino acid change within a variable region of gp120 which restored infectivity to near wild-type levels. These results, in combination with other point mutations that have been introduced into the HIV-1 envelope, suggest that at least three discrete regions of gp120 may interact during the establishment of a productive viral infection. This critical step occurs subsequent to the adsorption of virions to the cell surface and either prior to or concomitant with the fusion of viral and cellular membranes. PMID- 2547988 TI - Herpesvirus saimiri strains from three DNA subgroups have different oncogenic potentials in New Zealand white rabbits. AB - Herpesvirus saimiri is a primate tumor virus that induces acute T-cell lymphomas in New World monkeys. Strains of this virus have been previously classified into three groups on the basis of extreme DNA variability of the rightmost region of unique L-DNA. To compare the oncogenic potentials of various strains, we inoculated New Zealand White rabbits with viruses representing groups A, B, and C of herpesvirus saimiri. The results showed that a group C strain were highly oncogenic in New Zealand White rabbits; however, group A or B viruses were not oncogenic in these rabbits. Analysis of DNAs of tumor tissues and lymphoid cell lines established from tumors showed that the viral genome exists in circular episomal form. To identify which part of the genome of the group C strain is responsible for the highly oncogenic phenotype, group B-C recombinant strains were constructed by an efficient drug selection technique. Two group B recombinant strains in which the right-end 9.2 kilobase pairs of unique DNA is replaced by group C virus DNA were oncogenic in rabbits, indicating that the rightmost sequences contribute to the oncogenic properties of the group C strain. Oncogenicity of herpesvirus saimiri has been traditionally evaluated in New World monkeys; infection of rabbits with group C strain 484-77 offers a much more accessible animal model to study the mechanism of oncogenicity of this virus. PMID- 2547989 TI - Deletion analysis of the polyomavirus late promoter: evidence for both positive and negative elements in the absence of early proteins. AB - We have been interested in understanding more about the sequences that constitute the polyomavirus late promoter. Our approach has been to target specific deletions to the viral intergenic region by oligonucleotide-directed mutagenesis. Wild-type and mutant promoter cassettes with defined deletions were then inserted into a promoterless expression vector containing the bacterial chloramphenicol acetyltransferase (CAT) gene (cat). Plasmids were introduced into mouse NIH 3T3 cells by transfection, and promoter activities were assessed by quantitation of both CAT enzyme and cat mRNA levels. In this report, we present the results of experiments designed to map promoter elements which affect late transcription in the absence of early viral proteins and viral DNA replication. Using this approach, we mapped two major cis-acting elements (a positive and a negative one) which affect transcription in our transient expression system. The first, positive, element coincided with the enhancer A element, which is known to be important for early transcription and viral DNA replication. Removal of this element reduced late transcription by 50- to 100-fold. The second element was a negative one; removal of 89 base pairs that included two high-affinity large-T antigen-binding sites just to the early side of the inverted repeat structure within the replication origin resulted in a 5- to 10-fold increase in late promoter activity. The implications of these findings for late promoter function and regulation are discussed. PMID- 2547990 TI - Syncytium formation by recombinant vaccinia viruses carrying bovine parainfluenza 3 virus envelope protein genes. AB - The highly syncytium-inducing M strain and the weakly syncytium-inducing SC strain of bovine parainfluenza 3 virus differ by a single amino acid substitution in each of the hemagglutinin-neuraminidase (HN) and membrane (M) proteins, while their fusion (F) proteins are identical (T. Shioda, S. Wakao, S. Suzu, and H. Shibuta, Virology 162:388-396, 1988). We constructed recombinant vaccinia viruses which express separately the M virus HN (Vac-MHN), SC virus HN (Vac-SCHN), M virus M (Vac-MM), SC virus M (Vac-SCM), and common F (Vac-F) proteins. CV-1 cells were infected with the recombinants, singly or in combination, and implanted onto indicator MDBK cells for syncytium formation. Combinations of Vac-MHN plus Vac-F and Vac-SCHN plus Vac-F induced extensive and weak syncytium formation, respectively. Vac-F alone did not induce syncytium formation, and both Vac-MM and Vac-SCM had no effect on syncytium formation. These findings indicated that the syncytium formation by bovine parainfluenza 3 virus requires both the F and HN proteins and that the extensive syncytium formation by the M virus is due to the M virus HN protein. MSC, another weakly syncytium-inducing virus variant, newly isolated from the M virus, was identical to the M virus in the primary structure of the HN and M proteins but differed from the M virus by a single amino acid residue in the F protein. The combination of the recombinant vaccinia virus expressing the MSC virus F protein and Vac-MHN resulted in weak syncytium formation. PMID- 2547991 TI - Loss of responsiveness of an AP1-related factor, PEBP1, to 12-O tetradecanoylphorbol-13-acetate after transformation of NIH 3T3 cells by the Ha ras oncogene. AB - The function of the A element (nucleotides 5107 to 5130) of the polyomavirus enhancer is augumented in NIH 3T3 cells by a tumor-promoting phorbol ester, 12-O tetradecanoylphorbol-13-acetate (TPA). One of its targets is an AP1 consensus sequence motif recognized by a nuclear factor, PEBP1. In Ha-ras-transformed NIH 3T3 cells, however, A element function was not enhanced by TPA treatment, and at the same time PEBP1 was not detected in the nuclear extract by a mobility shift assay. PEBP1 was not detected in either the extract from NIH 3T3 cells treated in vivo with a protein kinase inhibitor, staurosporine, or the extract from NIH 3T3 cells after treatment in vitro with phosphatase. These results suggest that PEBP1 is required to be properly phosphorylated for DNA binding and that it is underphosphorylated, possibly due to the downregulation of protein kinase C in Ha ras-transformed cells. In addition, we observed that PEBP2, which bound to the A element adjacent to PEBP1, was converted to apparently related PEBP3 when conditions favored underphosphorylation. PMID- 2547992 TI - A subset of herpes simplex virus replication genes induces DNA amplification within the host cell genome. AB - Herpes simplex virus (HSV) induces DNA amplification of target genes within the host cell chromosome. To characterize the HSV genes that mediate the amplification effect, combinations of cloned DNA fragments covering the entire HSV genome were transiently transfected into simian virus 40 (SV40)-transformed hamster cells. This led to amplification of the integrated SV40 DNA sequences to a degree comparable to that observed after transfection of intact virion DNA. Transfection of combinations of subclones and of human cytomegalovirus immediate early promoter-driven expression constructs for individual open reading frames led to the identification of six HSV genes which together were necessary and sufficient for the induction of DNA amplification: UL30 (DNA polymerase), UL29 (major DNA-binding protein), UL5, UL8, UL42, and UL52. All of these genes encode proteins necessary for HSV DNA replication. However, an additional gene coding for an HSV origin-binding protein (UL9) was required for origin-dependent HSV DNA replication but was dispensible for SV40 DNA amplification. Our results show that a subset of HSV replication genes is sufficient for the induction of DNA amplification. This opens the possibility that HSV expresses functions sufficient for DNA amplification but separate from those responsible for lytic viral growth. HSV infection may thereby induce DNA amplification within the host cell genome without killing the host by lytic viral growth. This may lead to persistence of a cell with a new genetic phenotype, which would have implications for the pathogenicity of the virus in vivo. PMID- 2547993 TI - Identification of a domain required for autoproteolytic cleavage of murine coronavirus gene A polyprotein. AB - The 5'-most gene of the murine coronavirus genome, gene A, is presumed to encode viral RNA-dependent RNA polymerase. It has previously been shown that the N terminal portion of this gene product is cleaved into a protein of 28 kilodaltons (p28). To further understand the mechanism of synthesis of the p28 protein, cDNA clones representing the 5'-most 5.3 kilobases of murine coronavirus mouse hepatitis virus strain JHM were sequenced and subcloned into pT7 vectors from which RNAs were transcribed and translated in vitro. The sequence was found to encode a single long open reading frame continuing from near the 5' terminus of the genome. Although p28 is encoded from the first 1 kilobase at the 5' end of the genome, translation of in vitro-transcribed RNAs indicated that this protein was not detected unless the product of the entire 5.3-kilobase region was synthesized. Translation of RNAs of 3.9 kilobases or smaller yielded proteins which contained the p28 sequence, but p28 was not cleaved. This suggests that the sequence in the region between 3.9 and 5.3 kilobases from the 5' end of the genomic RNA is essential for proteolytic cleavage and contains autoproteolytic activity. The p28 protein could not be cleaved from the smaller primary translation products of gene A, even in the presence of the larger autocleaving protein. Cleavage of the p28 protein was inhibited by addition of the protease inhibitor ZnCl2. This study thus identified a protein domain essential for autoproteolytic cleavage of the gene A polyprotein. PMID- 2547994 TI - Identification of a new transcriptional initiation site and the corresponding functional gene 2b in the murine coronavirus RNA genome. AB - We have previously shown that some strains of the murine coronavirus mouse hepatitis virus (MHV) synthesize an additional mRNA species (mRNA 2b, previously called mRNA 2a) with a size intermediate between that of mRNAs 2 and 3, suggesting the presence of an optional transcriptional initiation site. This transcriptional start is dependent on the leader sequence of the virus strains. To study the mechanism of coronavirus transcriptional regulation, we have cloned and sequenced the region of the viral genome corresponding to the 5' unique coding region of mRNA 2 of the JHM strain of MHV. In addition to the open reading frame (ORF) predicted to encode the viral nonstructural protein p30, a second complete ORF, with the potential to encode a 439-amino-acid polypeptide, was discovered. The transcriptional initiation sites of both mRNA 2a (formerly called mRNA 2) and mRNA 2b were determined by primer extension studies and RNA sequencing. The data indicated that transcription of mRNA 2a initiated at a site, UCUAUAC, that resembled the consensus intergenic sequence. In contrast, the start signal of the optional mRNA 2b, UAAUAAAC, deviated from the consensus sequence. mRNA 2b is a functional mRNA, as shown by in vitro translation studies of mRNA and ORF 2b and by the detection of an additional viral structural protein, gp65, in the JHM strain that synthesized this mRNA. Although the A59 strain of MHV was found to retain ORF 2b, it lacked the correct transcriptional and translational start signals for this gene. This study has therefore identified an optional gene product for murine coronaviruses and provided insights into the mechanism of regulation of MHV RNA transcription. PMID- 2547995 TI - Herpes simplex virus ribonucleotide reductase: expression in Escherichia coli and purification to homogeneity of a tyrosyl free radical-containing, enzymatically active form of the 38-kilodalton subunit. AB - Infection of mammalian cells with herpes simplex virus (HSV) induces a virus encoded ribonucleotide reductase which is different from the cellular enzyme. This essential viral enzyme consists of two nonidentical subunits of 140 and 38 kilodaltons (kDa) which have not previously been purified to homogeneity. The small subunit of ribonucleotide reductases from other species contains a tyrosyl free radical essential for activity. We have cloned the gene for the small subunit of HSV-1 ribonucleotide reductase into a tac expression plasmid vector. After transfection of Escherichia coli, expression of the 38-kDa protein was detected in immunoblots with a specific monoclonal antibody. About 30 micrograms of protein was produced per liter of bacterial culture. The 38-kDa protein was purified to homogeneity in an almost quantitative yield by immunoaffinity chromatography. It contained a tyrosyl free radical which gave a specific electron paramagnetic resonance spectrum identical to that we have observed in HSV-infected mammalian cells and clearly different from that produced by the E. coli and mammalian ribonucleotide reductases. The recombinant 38-kDa subunit had full activity when assayed in the presence of HSV-infected cell extracts deficient in the native 38-kDa subunit. PMID- 2547996 TI - Identification and characterization of a human cytomegalovirus gene coding for a membrane protein that is conserved among human herpesviruses. AB - A rabbit antiserum was raised against envelope material from purified human cytomegalovirus strain AD169. The serum recognized polypeptides 200, 170, 160, 75, 58, and 45 kilodaltons in size. It was used to screen a cDNA library constructed from poly(A)+ RNA from human cytomegalovirus-infected cells in the expression vector lambda gt11. A recombinant bacteriophage expressing cytomegalovirus-specific sequences was identified, and the corresponding gene was mapped to the HindIII R fragment. The gene is transcribed into a late 1.5 kilobase RNA. The nucleotide sequence of the coding region was determined. Computer analysis of the gene product revealed a polypeptide containing multiple potential membrane-spanning domains, representing a type of protein not identified in the envelope of herpesviruses before. The protein shows homology on the amino acid level to hypothetical proteins from reading frames BBRF3 of Epstein-Barr virus, UL10 of herpes simplex virus type 1, and ORF50 of varicella zoster virus. By using an antiserum raised against procaryote-expressed parts of the cytomegalovirus membrane protein, a 45-kilodalton structural component of the virus was identified as the gene product. PMID- 2547998 TI - Helper-free stocks of recombinant adeno-associated viruses: normal integration does not require viral gene expression. AB - A method is described for the production of recombinant adeno-associated virus (AAV) stocks that contain no detectable wild-type helper AAV. The recombinant viruses contained only the terminal 191 nucleotides of the AAV chromosome bracketing a nonviral marker gene. trans-Acting AAV functions were provided by a helper DNA in which the terminal 191 nucleotides of the AAV chromosome were substituted with adenovirus terminal sequences. Although the helper DNA did not appear to replicate, it expressed AAV functions at a substantially higher level than did DNA molecules that contained neither AAV nor adenovirus termini. Since the recombinant viruses with AAV termini contained no sequence homology to the helper DNA, no wild-type AAV was generated by homologous recombination within infected cells. Since the terminal region of the AAV chromosome is required for replication and encapsidation, only recombinant DNAs were amplified and packaged into AAV virions. When human cells were infected at a high multiplicity with a recombinant virus carrying a drug resistance marker gene, approximately 70% of the infected cells gave rise to colonies stably expressing the marker. The recombinant virus gene was then used to generate drug-resistant human cell lines subsequent to infection. These cells contained stably integrated copies of the recombinant viral DNA which could be excised, replicated, and encapsidated by infection with wild-type AAV plus adenovirus. Thus, AAV gene expression is not required for normal integration of an infecting DNA containing AAV termini. PMID- 2547997 TI - Genetic identification of endogenous polytropic proviruses by using recombinant inbred mice. AB - Forty-seven endogenous polytropic murine viruses (Pmv) were identified by examination of proviral-cellular DNA junction fragment segregation in recombinant inbred (RI) mice. Most Pmv loci were found in more than one of the seven RI progenitor strains analyzed, but only four were present in all strains. Chromosomal assignments for 41 Pmv loci were determined by comparing their RI strain distribution patterns with those of known genetic markers. Pmv loci were found dispersed throughout the genome, with chromosomes 1, 3, 4, 5, 7, 11, 12, 15, and 16 each carrying three or more proviruses. Linkage analysis in the AKXD RI set suggested that the gene encoding mink cell focus-forming virus resistance (Rcmfr) of DBA/2J mice is probably not a Pmv provirus. It was also deduced that no single, AKR/J-specific Pmv provirus is required as an env gene donor for thymomagenic mink cell focus-forming viruses. In addition, a Pmv provirus was very closely associated with the albino mutation on chromosome 7. PMID- 2547999 TI - Cloning and expression of foreign genes in vaccinia virus, using a host range selection system. AB - A simple selection system has been developed for the cloning and expression of open reading frames in vaccinia virus. The selection system is based on a conditional lethal (host range) mutant of vaccinia virus. A deletion mutant of the vaccinia virus WR strain was generated by insertion of the neomycin resistance gene from transposon Tn5 and selection with the antibiotic G418. This deletion recombinant, vP293, lacked approximately 21.7 kilobases of DNA beginning 3.8 kilobases from the left end of the genome, vP293, was capable of plaquing on primary chicken embryo fibroblasts and two monkey cell lines (BSC-40 and Vero) but was defective in replication in the human cell line MRC-5. Insertion of the host range gene K1L into vP293 restored the ability to grow on MRC-5 cells. A series of plasmids were constructed which in addition to the K1L gene contained a vaccinia virus early-late promoter, H6, followed by a unique polylinker sequence, translational initiation and termination signals, and an early transcription termination signal. These plasmids, pHES1 through 4, allowed for rapid single step cloning and expression of any open reading frame when recombined in vivo with vP293 and scored for growth on MRC-5 cells. PMID- 2548000 TI - High recombination rate of an Epstein-Barr virus-simian virus 40 hybrid shuttle vector in human cells. AB - The stability of an Epstein-Barr virus (EBV)-simian virus 40 (SV40) hybrid shuttle vector, the p205-GTI plasmid, was analyzed in human cells during EBV- or SV40-type replication mode. When the p205-GTI plasmid was maintained as an episomal EBV vector in the human 293 cell line, no rearrangement was detected. To induce the SV40 replication mode, cells containing the episomal p205-GTI plasmid were either transfected with vectors carrying the T antigen gene or infected with SV40. Surprisingly, we observed both production and amplification of different classes of recombinant molecules. Particular types of modifications were found in most of the recombinants. The most striking rearrangement was a duplication of the promoter and enhancer regions of SV40 which was inserted in the thymidine kinase (TK) promoter. This recombination process involved a few bases of homology, and one of the recombination junctions implicated the GC boxes which constitute the essential components of the TK and SV40 early promoters. Our results suggest that a combination of a low level of base homology and a specific DNA sequence function (promoter and enhancer sites) leads to a very high level of recombinational activity during T-antigen-dependent plasmid replication. PMID- 2548001 TI - The 1:1 N-NS protein complex of vesicular stomatitis virus is essential for efficient genome replication. AB - We studied the effect pH had on the N-NS protein complex to determine its role in vesicular stomatitis virus (VSV) genome replication, as we had previously shown that VSV genome replication in vitro requires the interaction of the viral N and NS proteins into a 1:1 complex. A previous report showed that the growth of VSV in L cells was sensitive to the pH of the environment (M. Fiszman, J. B. Leaute, C. Chany, and M. Girard, J. Virol. 13:801-808, 1974). We hypothesized that low pH might disrupt the N-NS protein complex, and so we investigated the molecular events leading to inhibition of viral RNA replication in vitro from extracts that were prepared from VSV-infected cells incubated at pH 6.6. We found that viral genome RNA synthesis in vitro was reduced when infected cells were maintained at pH 6.6. Through immunoprecipitation analysis of the viral soluble protein pool, we found that a complex that usually exists between the N and NS proteins at pH 7.4 was altered in extracts from infected cells maintained at pH 6.6, and this was responsible for the observed effects on viral replication. The effect of low pH on the N-NS protein complex could not be abolished by increasing the concentration of the altered complex, indicating that the effects is more than simply a decrease in the level of the protein complex in the cell. Our data provide additional evidence that the 1:1 N-NS protein complex, and not the N protein alone, serves as the substrate for viral RNA replication in vivo. PMID- 2548003 TI - The Epstein-Barr virus BMLF1 promoter contains an enhancer element that is responsive to the BZLF1 and BRLF1 transactivators. AB - We have previously shown that the Epstein-Barr virus (EBV) immediate-early gene product, BZLF1, can activate expression of the EBV BMLF1 immediate-early promoter in EBV-positive, but not EBV-negative, B cells, suggesting that the BZLF1 effect may be mediated through another EBV gene product (S. Kenney, J. Kamine, E. Holley Guthrie, J.-C. Lin, E.-C. Mar, and J. S. Pagano, J. Virol. 63:1729-1736, 1989). Here, we show that the EBV BRLF1 immediate-early gene product transactivates the BMLF1 promoter in either EBV-positive or EBV-negative B cells. Deletional analysis revealed that both the BZLF1-responsive region and the BRLF1-responsive region of the BMLF1 promoter are contained within the same 140-base-pair FokI PvuII fragment located 300 base pairs upstream of the mRNA start site. This FokI PvuII fragment functions as an enhancer element in the presence of the BRLF1 transactivator and contains the sequence CCGTGGAGA ATGTC, which is strikingly similar to the BRLF1-responsive region of the EBV DR/DL enhancer (A. Chevallier Greco, H. Gruffat, E. Manet, A. Calender, and A. Sergeant, J. Virol. 63:615-623, 1989). The effect of BZLF1 on the BMLF1 promoter is likely to be indirect and mediated through the BRLF1 transactivator. PMID- 2548002 TI - The Epstein-Barr virus immediate-early gene product, BMLF1, acts in trans by a posttranscriptional mechanism which is reporter gene dependent. AB - In DNA cotransfection experiments, the Epstein-Barr virus immediate-early gene product, BMLF1, stimulated the chloramphenicol acetyltransferase (CAT) activity of both latent and productive EBV promoters linked to CAT. This BMLF1-induced increase in CAT activity was out of proportion to the effect on CAT mRNA, suggesting a posttranscriptional mechanism. Furthermore, when growth hormone was used as a reporter gene instead of CAT, BMLF1 no longer functioned. Thus, the BMLF1 effect was reporter-gene dependent. The effect of the BMLF1 gene product does not then appear to be directed at promoter activation, but instead may function to increase the level of an as yet unidentified protein(s) required for Epstein-Barr virus infection. PMID- 2548004 TI - Leader-encoded open reading frames modulate both the absolute and relative rates of synthesis of the virion proteins of simian virus 40. AB - Numerous viral and cellular RNAs are polycistronic, including several of the late mRNA species encoded by simian virus 40 (SV40). The functionally bicistronic major late 16S and functionally tricistronic major late 19S mRNA species of SV40 contain the leader-encoded open reading frames (ORFs) LP1, located upstream of the sequence encoding the virion protein VP1, and LP1*, located upstream of the sequence encoding the virion proteins VP2 and VP3. To determine how these leader ORFs affect synthesis of the virion proteins, monkey cells were transfected with viral mutants in which either the leader-encoded translation initiation signal was mutated or the length and overlap of the leader ORF relative to the ORFs encoding the virion proteins were altered. The levels of initiation at and leaky scanning past each initiation signal were determined directly by quantitative analysis of the viral proteins synthesized in cells transfected with these mutants. Novel findings from these experiments included the following. (i) At least one-third of ribosomes bypass the leader-encoded translation initiation signal, GCCAUGG, on the SV40 major late 16S mRNA. (ii) At least 20% of ribosomes bypass even the consensus translation initiation signal, ACCAUGG, when it is situated 10 bases from the 5' end on the major late 16S mRNA. (iii)O The presence of the leader ORF on the bicistronic 16S mRNA species reduces VP1 synthesis threefold relative to synthesis from a similar RNA that lacks it. (iv) At least half and possibly all VP1 synthesized from the bicistronic 16S mRNA species is made by a leaky scanning mechanism. (v) LP1 and VP1 are synthesized from the bicistronic 16S mRNA species at approximately equal molar ratios. (vi) Approximately half of the VP1 synthesized in SV40-infected cells is synthesized from the minor, monocistronic 16S mRNA even though it accounts for only 20% of the 16S mRNA present. (vii) The presence and site of termination of translation of the leader ORF on the late 19S mRNAs affect the relative as well as absolute rates of synthesis of VP2 and VP3. PMID- 2548005 TI - Phosphorylation of varicella-zoster virus glycoprotein gpI by mammalian casein kinase II and casein kinase I. AB - Varicella-zoster virus (VZV) glycoprotein gpI is the predominant viral glycoprotein within the plasma membranes of infected cells. This viral glycoprotein is phosphorylated on its polypeptide backbone during biosynthesis. In this report, we investigated the protein kinases which participate in the phosphorylation events. Under in vivo conditions, VZV gpI was phosphorylated on its serine and threonine residues by protein kinases present within lysates of either VZV-infected or uninfected cells. Because this activity was diminished by heparin, a known inhibitor of casein kinase II, isolated gpI was incubated with purified casein kinase II and shown to be phosphorylated in an in vitro assay containing [gamma-32P]ATP. The same glycoprotein was phosphorylated when [32P]GTP was substituted for [32P]ATP in the protein kinase assay. We also tested whether VZV gpI was phosphorylated by two other ubiquitous mammalian protein kinases- casein kinase I and cyclic AMP-dependent kinase--and found that only casein kinase I modified gpI. When the predicted 623-amino-acid sequence of gpI was examined, two phosphorylation sites known to be optimal for casein kinase II were observed. Immediately upstream from each of the casein kinase II sites was a potential casein kinase I phosphorylation site. In summary, this study showed that VZV gpI was phosphorylated by each of two mammalian protein kinases (casein kinase I and casein kinase II) and that potential serine-threonine phosphorylation sites for each of these two kinases were present in the viral glycoprotein. PMID- 2548006 TI - Characterization of hepatitis B virus major surface antigen gene transcriptional regulatory elements in differentiated hepatoma cell lines. AB - The regulatory DNA sequence elements that control the expression of the hepatitis B virus major surface antigen gene in the hepatoblastoma cell line HepG2 were analyzed by using transient transfection assays. In this system, the hepatitis B virus enhancer increases transcription from the surface antigen promoter approximately twofold. The promoter elements regulating the expression of this gene are within a 200-nucleotide sequence located immediately upstream of the transcription initiation sites. The promoter consists of an 85-nucleotide distal element which increases transcription from the surface antigen gene by two- to fourfold and a proximal element of approximately 115 nucleotides which is essential for transcriptional activity. The proximal and distal promoter elements were shown to bind factors present in HepG2 nuclear extracts, which is consistent with the regulatory role demonstrated for these sequences. The regulatory role of these promoter sequences in the hepatocellular carcinoma cell lines PLC/PRF/5 and Hep3B was also demonstrated, indicating similar transcriptional regulation of the surface antigen gene in each of these differentiated hepatoma cell lines. PMID- 2548007 TI - Severe thrombocytopenia in young calves experimentally infected with noncytopathic bovine viral diarrhea virus. AB - Seven calves between 1 week and 2 months of age were infected with a noncytopathic field isolate of bovine viral diarrhea virus (BDV) in order to evaluate the effect of BDV infection on the concentration of circulating platelets in the blood. All calves were determined to be free of BDV and neutralizing antibodies to BDV before infection. Platelet counts were performed on a daily basis over a 30-day period beginning at the time of infection. By 2 weeks postinfection, all calves showed a significant drop in the number of circulating platelets and a marked hyperplasia of megakaryocytes in the bone marrow. In three of the seven calves, thrombocytopenia was severe (less than or equal to 5,000/microliters) for 1 to 6 days. In two of these three animals, extensive petechial and ecchymotic hemorrhages were observed on all mucosal surfaces and on various internal organs during the period of severe thrombocytopenia. BDV was consistently isolated from the platelets during the early phases of the infection, and viral antigen was occasionally detected on platelets by a fluorescent-antibody assay. The results demonstrate that BDV infection is associated with decreases in platelet numbers and suggest that platelets may serve as carriers of circulating virus. PMID- 2548008 TI - Oncogenicity of human N-ras oncogene and proto-oncogene introduced into retroviral vectors. AB - The N-ras gene is the only member of the ras family which has never been naturally transduced into a retrovirus. In order to study the in vitro and in vivo oncogenicity of N-ras and to compare its pathogenicity to that of H-ras, we have inserted an activated or a normal form of human N-ras cDNA into a slightly modified Harvey murine sarcoma virus-derived vector in which the H-ras p21 coding region had been deleted. The resulting constructions were transfected into NIH 3T3 cells. The activated N-ras-containing construct (HSN) induced 10(4) foci per microgram of DNA and was found to be as transforming as H-ras was. After infection of the transfected cells by either the ecotropic Moloney murine leukemia virus or the amphotropic 4070A helper viruses, rescued transforming viruses were injected into newborn mice. Both pseudotypes of HSN virus containing activated N-ras induced the typical Harvey disease with similar latency. However, we found that the virus which contained normal N-ras p21 (HSn) was also pathogenic and induced splenomegaly, lymphadenopathies, and sarcoma in mice after a latency of 3 to 7 weeks. In addition, Moloney murine leukemia virus pseudotypes of N-ras caused neurological disorders in 30% of the infected animals. These results differed markedly from those of previous experiments in which we had inserted the activated form of N-ras in the pSV(X) vector: the resulting SVN-ras virus was transforming on NIH 3T3 cells but was poorly oncogenic in vivo (M. Souyri, C. F. Koehne, P. V. O'Donnel, T. H. Aldrich, M. E. Furth, and E. Fleissner, Virology 158:69-78). However, similarly poor oncogenicity was also observed when the v-H-ras coding sequence was inserted in pSV(X) vector, which indicated that the vector sequences play a crucial role in the pathogenicity of a given oncogene. Altogether, these data demonstrated unequivocally that N-ras is potentially as oncogenic as H-ras and that such oncogenic effect could depend on the vector environment. PMID- 2548009 TI - Poliovirus-associated protein kinase: destabilization of the virus capsid and stimulation of the phosphorylation reaction by Zn2+. AB - The previously described poliovirus-associated protein kinase activity phosphorylates viral proteins VP0 and VP2 as well as exogenous proteins in the presence of Mg2+. In this paper, the effect of Zn2+ on the phosphorylation reaction and the stability of the poliovirus capsid has been studied in detail and compared to that of Mg2+. Phosphorylation patterns of viral and other proteins depend on the divalent cation present. In the presence of Zn2+, phosphorylation of capsid proteins VP2 and VP4 is significantly higher while phosphorylation of VP0 and exogenous phosphate acceptor proteins is not detected. Our results indicate the activation of more than one virus-associated protein kinase by Zn2+. The ion-dependent behavior of the enzyme activities is observed independently of whether the virus was obtained from HeLa or green monkey kidney cells. The poliovirus capsid is destabilized by Zn2+. The destabilization leads to a substantially increased permeability of virus particles to ethidium bromide and RNase, concomitant with decreased infectivity of the sample. This alteration of the poliovirus capsid structure is a prerequisite for effective phosphorylation of viral capsid proteins. The increased level of phosphorylation of viral capsid proteins results in further destabilization of the viral capsid. As a result of the conformational changes, poliovirus-associated protein kinase activities dissociate from the virus particle. High-performance liquid chromatography-purified viral protein VP2 is phosphorylated by the released enzymes on serine, threonine, and tyrosine in the presence of Zn2+. We suggest that the destabilizing effect of phosphorylation on the viral capsid plays a role in uncoating of poliovirus. PMID- 2548010 TI - Rotavirus SA11 genome segment 11 protein is a nonstructural phosphoprotein. AB - We investigated properties of the rotavirus genome segment 11 protein. A rotavirus SA11 genome segment 11 cDNA which contains the entire coding region was sequenced and inserted into the baculovirus transfer vector pVL941. Recombinants containing gene 11 cDNA were selected, and the gene 11 product expressed in Spodoptera frugiperda cells infected with these recombinants was inoculated into guinea pigs to produce hyperimmune antiserum. Characterization of the antiserum showed that it recognized a primary translation product with a molecular weight of 26,000 (26K protein) in recombinant-infected insect cells, in SA11-infected monkey kidney cells, and in cell-free translation reactions programmed with SA11 mRNA. A modified 28K product was also detected but only in SA11-infected monkey kidney cells. The 26K 28K proteins were shown to be phosphorylated in infected monkey kidney cells, and the 26K protein was phosphorylated in insect cells. We were unable to identify what type of modification caused the molecular weight shift to 28,000 in infected monkey kidney cells. Large amounts of the gene 11 product were detected by immunofluorescence in discrete foci in the cytoplasm of infected monkey kidney cells. Viruses of all known serotypes were also detected by immunofluorescence by using hyperimmune antiserum to the SA11 gene 11 product. The antiserum reacted with particle-depleted cytosol fractions but did not react with purified virus particles by immunoprecipitation or immunoblotting; it also did not neutralize virus infectivity in plaque reduction neutralization assays. Therefore, we conclude that the primary gene 11 product is a nonstructural phosphoprotein which we designated NS26. PMID- 2548011 TI - Human cytomegalovirus binding to fibroblasts is receptor mediated. AB - The binding of radiolabeled human cytomegalovirus (HCMV) strain AD169 to human lymphocytes, lymphoblastoid cell lines, monocytes, and fibroblasts varied over a 20-fold range. Since maximum binding was observed with human foreskin fibroblasts (HFF), interactions of radiolabeled HCMV with this cell type were analyzed quantitatively. Binding of HCMV to HFF at 4 degrees C was specific and saturable; at low viral inputs specific binding averaged 16.4% of input and nonspecific binding was less than 1% of input. Binding curves yielded single-component linear Scatchard plots indicating an average Kd of 1.1 nM and 5,262 available virus binding sites per cell. A two-component Scatchard curve was obtained at 37 degrees C and reflected viral internalization, since it could be converted to a single-component curve by the use of paraformaldehyde-fixed cells. HCMV strain Towne was found to bind to the receptor used by HCMV strain AD169 with similar affinity. HCMV failed to bind to protease-treated HFF or to HFF grown in the presence of inhibitors of glycosylation. Sialic acid residues, however, were not found to be important in binding. These data indicate that a single type of molecule, likely a glycoprotein, on the surface of HFF serves as a specific receptor for the virus. PMID- 2548012 TI - Effect of infection with the ts22 mutant of Semliki Forest virus on development of the central nervous system in the fetal mouse. AB - The A7 strain of Semliki Forest virus induces rapid fetal death in pregnant mice, whereas the ts22 mutant derived from it is teratogenic for a proportion of fetuses. Both A7 and ts22 induce viremia and infect the central nervous systems and fetuses of pregnant mice. Using immunogold-silver staining, a cDNA probe for a Semliki Forest virus nonstructural sequence, and a riboprobe derived from the same sequence, we showed that the skin and musculoskeletal systems of fetuses from mothers infected with ts22 were often heavily infected but the central nervous systems were not labeled before day 17 of pregnancy. Damage to the neural tube, including open-neural-tube defects, was detected in fetuses following infection of the mother at days 8 and 10 of pregnancy with both A7 and ts22. For ts22, neural tube damage induced by fetal infection before day 17 of pregnancy appeared to be indirect and caused by virus infection of mesenchymal cells surrounding the developing neural tube. PMID- 2548013 TI - Restricted growth of attenuated poliovirus strains in cultured cells of a human neuroblastoma. AB - Cultured cells of a human neuroblastoma, SK-N-MC, were found to be highly resistant to Sabin attenuated poliovirus types 1 and 2 strains; no appreciable cytopathic effect was observed, and the total harvest was generally in the order of 1 PFU per cell or less. On the other hand, related neurovirulent strains of these antigenic types produced a relatively good (2 orders of magnitude higher) yield in a markedly protracted infectious cycle. The limited growth of the attenuated virus in the neuroblastoma cells appeared to be confined to a minor cell subpopulation. Experiments with intratypic (type 1) poliovirus recombinants suggested that the major genetic determinants limiting reproduction of the attenuated polioviruses in the neuroblastoma cells are located in the 5' half of the viral RNA, although the 3' half also appears to contribute somewhat to this phenotype. The possibility that neuroblastoma cells may represent an in vitro model for studying poliovirus neurovirulence is briefly discussed. PMID- 2548014 TI - An amplified endogenous retroviral sequence on the murine Y chromosome related to murine leukemia viruses and viruslike 30S sequences. AB - A highly repeated sequence on the murine Y chromosome was cloned and characterized. The DNA sequence of the viral long terminal repeats (LTRs) showed that the 5' and 3' LTRs were approximately 90% homologous. The LTRs are generally unrelated to any previously reported viral LTR but are somewhat similar to the viruslike 30S sequences. PMID- 2548015 TI - Replacement of the pseudorabies virus glycoprotein gIII gene with its postulated homolog, the glycoprotein gC gene of herpes simplex virus type 1. AB - gIII, the major envelope glycoprotein of pseudorabies virus (PRV), shares approximately 20% amino acid similarity with glycoprotein gC of herpes simplex virus type 1 (HSV-1) and HSV-2. We describe here our first experiments on the potential conservation of function between these two genes and gene products. We constructed PRV recombinants in which the gIII gene and regulatory sequences have been replaced with the entire HSV-1 gC gene and its regulatory sequences. The gC promoter functions in the PRV genome, and authentic HSV-1 gC protein is produced, albeit at a low level, in infected cells. The gC protein is present at the cell surface but cannot be detected in the PRV envelope. PMID- 2548016 TI - Epstein-Barr virus latent infection membrane protein increases vimentin expression in human B-cell lines. AB - Latent Epstein-Barr virus (EBV) infection activates B-lymphocyte proliferation through mechanisms which are partially known. One approach to further delineate these mechanisms is to identify cellular genes whose expression is augmented in cells latently infected with EBV. Since EBV-negative Burkitt's lymphoma cells can be grown in continuous culture and EBV can establish growth-altering latent infection in these cells, some effects of EBV on B-lymphocyte gene expression can be studied by using this in vitro system. Pursuing this latter approach, we have used cDNA cloning and subtractive hybridization to identify a gene whose expression is increased after EBV infection. This gene encodes the cytoskeletal protein vimentin. Latent infection of established EBV-negative Burkitt's lymphoma cell lines with the transforming EBV strain, B95-8, resulted in dramatic increases in vimentin mRNA and protein levels, while infection with the nontransforming P3HR1 strain failed to do so. Vimentin induction was reproduced by the expression of the single EBV gene which encodes the latent infection membrane protein (LMP). An amino-terminal LMP deletion mutant did not induce vimentin. These results are of particular interest in light of the transforming potential of LMP, as demonstrated in rodent fibroblasts, and the interaction between vimentin and LMP observed in immunofluorescent colocalization and cell fractionation studies. PMID- 2548018 TI - Reduced reproductive capacity of freshly harvested endothelial cells in smokers: a possible shortcoming in the success of seeding? AB - In an attempt to explain the failure of first clinical trials of autologous endothelial seeding in smokers, the initial reproductive capacity of saphenous vein endothelial cells from smokers and nonsmokers was studied by a replicate microwell technique. Endothelial cells were enzymatically harvested from saphenous vein segments of patients with coronary bypasses (21 smokers and 18 nonsmokers). After 15 minutes (group A) and 7 minutes (group B) of collagenase exposure, the endothelial cell harvest from donors who smoked was 41% (p less than 0.02) lower for group A and 30% (p less than 0.2) lower for group B than that from nonsmokers. In analogy, the viable cell yield was 32% (p less than 0.04) and 29% (p less than 0.05) lower for groups A and B, respectively, in cultures from donors who smoked. Daily cell counts over an ensuing 10-day period also revealed a significant difference in the proliferative behavior of endothelial cells from smokers and nonsmokers. Whereas endothelial cells from nonsmokers regularly entered the exponential phase of proliferation on day 4.4 +/ 1.8 (group A) and day 4.6 +/- 1.3 (group B), endothelial cells from smokers reached the logarithmic growth phase either with delay (day 6.8 +/- 2.1, group A) or remained completely quiescent (group B). Lower harvest efficiency and suppressed reproductive capacity of endothelial cells in smokers--on top of an already critically low inoculum in single-staged endothelial cell seeding--might explain the failure of initial clinical trials. PMID- 2548017 TI - Examination of the roles of transcription factor Sp1-binding sites and an octamer motif in trans induction of the herpes simplex virus thymidine kinase gene. AB - Herpes simplex virus mutants with both Sp1-binding sites in the thymidine kinase (tk) promoter inactivated or an octamer motif deleted were at most modestly impaired for tk expression. Thus, no cellular transcription factor that binds upstream of the tk TATA box is solely required for trans induction of this gene. PMID- 2548019 TI - Observations on human smooth muscle cell cultures from hyperplastic lesions of prosthetic bypass grafts: production of a platelet-derived growth factor-like mitogen and expression of a gene for a platelet-derived growth factor receptor--a preliminary study. AB - Prosthetic bypass grafts placed to the distal lower extremity often fail because of an occlusive tissue response in the perianastomotic region. The origin of the cells that comprise this occlusive lesion and the causes of the cellular proliferation are not known. To increase our understanding of this process we cultured cells from hyperplastic lesions obtained from patients at the time of reexploration for lower extremity graft failure, and we studied their identity and growth factor production in tissue culture. These cultures contain cells that express muscle-specific actin isoforms, shown by immunohistochemical staining, consistent with vascular smooth muscle origin. These cultures also released material that stimulated smooth muscle cell growth. A portion of this activity was similar to platelet-derived growth factor, since preincubation with antibody to-human platelet-derived growth factor partially blocked the mitogenic effect of medium conditioned by human anastomotic hyperplastic cells. These conditioned media also contained material that competed with platelet-derived growth factor for its receptor, as measured in a radioreceptor assay. Northern blot analysis showed that these cells contain messenger RNA that encodes the A chain but not the B chain of platelet-derived growth factor. In addition, these cells contain messenger RNA that encodes a platelet-derived growth factor receptor. We conclude that cultured smooth muscle cells from human anastomotic hyperplastic lesions express genes for platelet-derived growth factor A chain and a platelet-derived growth factor receptor and secrete biologically active molecules similar to platelet-derived growth factor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548020 TI - Long past date set for its US disappearance, measles remains a threat to many children. PMID- 2548021 TI - Exacerbation of hypercapnia and acidosis of central venous blood and tissue following administration of sodium bicarbonate during cardiopulmonary resuscitation. AB - Administration of sodium bicarbonate during cardiopulmonary resuscitation (CPR) is controversial, and our aim was to elucidate whether or not its administration is beneficial by analyzing the acid-base status and the level of carbon dioxide in central venous blood during CPR, and their changes following administration of sodium bicarbonate. Six patients were studied. They had all been admitted to the intensive care unit (ICU), had already had pulmonary arterial or central venous catheters inserted, and had acute episodes of circulatory collapse during their stay in the ICU. The following phenomena were observed: 1) hypercapnia and acidosis of central venous blood were prominent during both cardiogenic shock and CPR, although arterial hypocapnia was maintained by hyperventilation; 2) administration of sodium bicarbonate during cardiogenic shock and CPR induced exacerbation of hypercapnia and acidosis of central venous blood; 3) when arterial hypercapnia was present due to disturbed ventilation, administration of sodium bicarbonate exacerbated hypercapnia and acidosis of both arterial and central venous blood; 4) administration of sodium bicarbonate did not induce hypercapnia of central venous blood in a septic shock patient in whom the septic hyperdynamic state was prevalent in spite of low systemic perfusion pressure. It was concluded that hypercapnia and acidosis of the central venous blood and tissues were exacerbated by administration of sodium bicarbonate during CPR, and that such an effect might be dependent on the severity of the decrease in tissue perfusion. PMID- 2548022 TI - [My experience in child birth in England. 7. The care and support system following discharge from the hospital]. PMID- 2548023 TI - Activation by systemic GABA of vagal efferent transmission in the rat: correlation to its acid secretagogue action. AB - Stimulatory effects of systemically administered gamma-aminobutyric acid (GABA) on gastric acid secretion and vagal efferent activity were studied in anesthetized rats. Intravenous injection of GABA (400 mg/kg) significantly increased gastric acid secretion. The secretagogue effect of GABA on the gastric acid secretion was partially attenuated by vagotomy by approximately 60%. Atropine (1 mg/kg, s.c.) completely abolished the stimulatory effect of GABA on the acid output. GABA, at secretagogue doses, enhanced the firing rate of vagal efferent activity. These results suggest that the secretagogue action of intravenously injected GABA is primarily mediated by muscarinic mechanisms and that the stimulation of the central vagal efferent pathway might trigger the excitatory action of systemic GABA. PMID- 2548024 TI - Morphologic characteristics of a transplantable tumor derived from a spontaneous malignant fibrous histiocytoma in the rat. AB - Malignant fibrous histiocytoma (MFH) developed spontaneously in subcutaneous tissue of the head of a 15-month-old male Fischer 344 rat. The tumor was serially transplanted into syngeneic rats up to the 45th generation and was designated MFH MT. Light and electron microscopic examinations revealed that the original and serially transplanted tumors were composed of an admixture of fibroblast-like and histiocyte-like cells arranged in a storiform pattern. Neoplastic cells gave positive reactions for acid phosphatase, alkaline phosphatase, nonspecific esterase, alpha-1 antitrypsin and lysozyme. The tumors transplanted into the lungs and cutaneous tissue of the tail had a mixed histologic appearance of storiform, pleomorphic, myxoid and giant cell types. Moreover sclerosing hemangioma-like and osteosarcoma-like structures were also found. MFH-MT grew well in athymic nude mice showing neoplastic proliferation of pleomorphic cells strongly positive for alpha-1 antitrypsin. Development of MFH-MT was significantly retarded by the two antitumor drugs tested. The retarded tumors consisted predominantly of fibroblast-like cells and abundant collagenic fibers, whereas histiocytic cells decreased in number. PMID- 2548025 TI - Escherichia coli associated endotoxemia in dogs with parvovirus infection. AB - Escherichia coli bacteremia and endotoxemia were observed in 3 adult mongrel dogs which had been prediagnosed as canine parvoviral disease. The endotoxin level was 46.5 pg/ml in the plasma of clinical cases, while 2.3 pg/ml in healthy controls. The microflora of the feces was confused in the clinical cases. The percentage of E. coli was major in the feces. Serologically similar strains were isolated from the blood. These strains did not produce enterotoxins such as heat-stable enterotoxin (ST) and heat-labile enterotoxin (LT). Histopathologically, the lesions in the small intestine consisted of epithelial degeneration and necrosis. Viral inclusion bodies were frequently observed in the epithelial cells. Disseminated intravascular coagulation was observed in various tissues including the liver and small intestinal submucosa. After experimental infection with CPV, all dogs showed various clinical signs. CPV was positive in the feces. Endotoxin level in the plasma gradually increased and high level continued for long period from 10 to 30 days. Mean maximum level of endotoxin in the experimental dogs was 73.6 pg/ml. These results indicate that intestinal flora plays a important role in the pathogenesis of CPV infection and that endotoxin is one of the factors which predispose to severe disease after the infection. PMID- 2548026 TI - Infectivity of infectious bursal disease virus neutralized by maternal antibody in various chicken cells. PMID- 2548027 TI - Seroepizootiologic survey of feline syncytial virus infections in domestic cats. PMID- 2548028 TI - Isolation of reovirus type 3 from foals. PMID- 2548029 TI - Some attributes of a rotavirus detected in cat feces. PMID- 2548030 TI - Comparison of physicochemical properties of mouse hepatitis virus strains. PMID- 2548031 TI - [Biogenesis of the insulin secretion granule]. PMID- 2548032 TI - [Pathways of insulin secretion: the role of calcium and other intracellular messengers]. PMID- 2548033 TI - [Effect of taurine on cytogenetic disorders in murine cornea caused by 9GeV proton irradiation]. PMID- 2548034 TI - The use of dirty bedding for detection of murine pathogens in sentinel mice. AB - Sentinel Swiss (CD-1) mice, housed without filter bonnets, were seronegative for mouse hepatitis virus (MHV) for 8 consecutive months in an experimental colony of CD-1 mice. MHV titers had been detected sporadically in sentinel mice housed in this colony during a 2 year period. In an effort to determine whether MHV was still present in the colony, two methods of exposing sentinel mice to an animal room environment were compared under routine husbandry practices. Eight cages (12 mice per cage; 2 cages per rack) of experimental virus antibody free sentinel mice, housed without filter bonnets, were placed on the bottom shelf of 4 of 12 racks in the room. Twice each week, four cages of sentinel mice received a composite sample of dirty bedding (bedding used previously by mice in the room). The remaining four cages of experimental sentinels received fresh non-used bedding. Sentinel mice were bled at monthly intervals for MHV serology. After 4 months, mice from two cages which received dirty bedding seroconverted to MHV and mice from one cage were positive for Myobia musculi (mites). Three weeks later, all four cages of mice which received dirty bedding were positive for MHV and three were positive for mites. In contrast, only two of four cages of mice which received fresh bedding were positive for MHV and all were negative for mites. These findings indicate the importance of exposing sentinel mice to dirty bedding and that MHV and mites may go undetected for several months in a mouse colony when the incidence levels are low where standard sanitation procedures are used. PMID- 2548035 TI - Infection of rabbits with Sendai virus. AB - Rabbits were either inoculated with Sendai virus (SV), strain MN, or caged with virus-inoculated rabbits on the same day of the viral inoculation, and examined for viral shedding and detection of viral antigens in the respiratory tract, histopathologic changes, and serum antibodies. Infectious virus was recovered from nasal swabs at postinoculation day (PID) 3 and disappeared by PID 10. Viral antigens were detected by immunofluorescence in epithelial cells of the nasal cavities, but not of the trachea and lungs from PID 3 to PID 10, and antibodies were detected after PID 7. Rabbits had no clinical manifestations and only exhibited a moderate increase in goblet cells of the nasal epithelium. In the transmission study, virus was recovered from one of three uninoculated rabbits at postexposure day (PED) 10 and antibodies were detected at PED 15 in the same rabbit. These data suggest that, although viral multiplication was limited to the nasal epithelium, laboratory rabbits are susceptible to Sendai virus infection. PMID- 2548036 TI - Diagnostic exercise: fetal death in guinea pigs. PMID- 2548037 TI - Influence of kaurenol on basal, LH- and forskolin-stimulated progesterone and cAMP production in avian granulosa cells. AB - The effects of kaurenol, a diterpene alcohol, were evaluated on progesterone and cyclic AMP (cAMP) production in freshly dispersed avian granulosa cells. Kaurenol (50 microM) alone caused a fourfold increase in progesterone synthesis without a measurable influence on cAMP levels. When granulosa cells were challenged with near-maximally stimulating concentrations of LH (50 ng/ml) or forskolin (10 microM), kaurenol (10-100 microM) dose-dependently suppressed steroidogenesis. Similarly, cAMP production in response to LH and forskolin stimulation was also inhibited. When progesterone synthesis was stimulated by the addition of pregnenolone or 25-hydroxycholesterol substrates to the culture medium, the typical dose response to the latter precursor, but not to pregnenolone, was abolished by kaurenol. Whereas the mechanism of kaurenol's stimulatory effect on basal steroidogenesis remains unknown, it is suggested that its inhibitory action on LH- and forskolin-promoted progesterone production may be due to the inhibition of the adenylate cyclase cAMP effector system as well as to the impairment of the action of the mitochondrial cholesterol side chain cleavage enzyme system. PMID- 2548038 TI - Comparison of the effects between the conventional and electrical acupuncture on some cytochemical components of median eminentia and supraoptic nucleus. PMID- 2548039 TI - Effect of acupuncture on left ventricular function, microcirculation, blood rheology and cyclicnucleotides in patients with acute myocardial infarction. PMID- 2548041 TI - Characterization of cell markers in type B retroviral-induced thymic lymphomas- II. Surface antigen phenotype, karyotype and proviral integration pattern in cultured lymphoma cells and cloned lines. AB - This study examined the surface antigen phenotype, karyotype and proviral integration patterns of cultured cells from murine thymic lymphomas induced by injecting neonatal mice with the type B leukemogenic retrovirus (DMBA-LV). Cells from the primary thymic lymphomas were established in mass culture and from these, clonal tumor cell lines were derived. During in vitro culture of lymphoma cells, Lyt 1-2+ cells predominated with an apparent selection against cells of the Lyt 1+2- and Lyt 1+2+ phenotypes. Of 21 cloned lines established, five had a diploid chromosome complement and expressed the Lyt 1-2+ phenotype. The other 16 clones lines were characterized by trisomy of chromosome 15 and expressed the Lyt 1+2+ or Lyt 1-2+ phenotype. Cells characterized by either a diploid or trisomy chromosome complement were capable of growth in vivo. Southern blot analyses showed that during growth in culture, cells from the mass cultures and cloned lines continued to acquire low numbers of new integrated DMBA-LV proviral copies while maintaining the basic proviral integration pattern present in the DNA from cells of the primary lymphomas. These findings support the notion that the acquisition of new genetic information in cells from DMBA-LV-induced thymic lymphomas may contribute to the continual generation of tumor heterogeneity. PMID- 2548040 TI - Characterization of cell markers in type B retroviral-induced thymic lymphomas- I. Surface antigen phenotype and karyotype in developing and primary lymphomas. AB - CFW/D mice injected neonatally with a type B retrovirus, (DMBA-LV), rapidly develop thymic lymphomas. The present study examined simultaneously the karyotype and the expression of surface antigens on thymocytes from DMBA-LV treated mice at 28, 35 and 42 days of age during the development of lymphomas, and of cells from primary lymphomas. DMBA-LV treatment resulted in disturbances in the proportions of thymic Lyt 1+2-, Lyt 1-2+ and Lyt 1+2+ subpopulations. As a group, virus treated mice showed a significant decrease in the size of the Lyt 1+2- subpopulation in thymuses during tumor development. However, developing tumors and primary tumors showed individual patterns of alteration in the proportions of thymocyte subpopulations rather than consistent trends for any one thymocyte subpopulation to increase or decrease. Cells with the abnormal chromosome complement, trisomy 15, were present early in the development of tumors and became the predominant karyotype in fully developed tumors. A correlation between the appearance of trisomy 15 and a particular thymocyte surface antigen phenotype was not detected. The results suggest that retrovirus infection disrupts, but does not eliminate the ability of developing thymocytes to form phenotypically distinct subpopulations. PMID- 2548042 TI - The DNA synthesis of leukemic (L2C) guinea pig B lymphocytes involves a permanent activation of protein kinase C without corresponding phosphoinositide hydrolysis. AB - L2C B lymphocytes have a constant high DNA synthesis due to their continuous proliferative state. The addition of polymyxin B (PmB), a rather selective inhibitor of protein kinase C, stopped (3H)thymidine incorporation with an IC50 of 10 microM when added 18 h before measuring DNA synthesis. Interestingly, PmB inhibition of DNA synthesis was suppressed when 4 nM 12-O-tetradecanoylphorbol-13 acetate was added along with PmB, indicating that PmB may act through inhibition of protein kinase C. In the node and spleen lymphocytes of normal guinea pigs, protein kinase C activity was entirely cytosolic and was eluted at 0.12 M NaCl when adsorbed on DEAE-cellulose. In L2C leukemic lymphocytes, total protein kinase C activity was of the same order of magnitude, but 20% of it was associated with the membrane fraction. The lipid-dependent activity, eluted at 0.12 M NaCl from cytosolic and membrane fractions, was suppressed by staurosporine with an IC50 of 10-40 nM and by polymyxin B with an IC50 of 2-6 microM. Phosphoinositide metabolism was studied in the transformed cells. Incorporation of 32Pi into polyphosphoinositides was considerable, whereas much more time was required for a tiny incorporation of inositol. We detected no release of radioactive inositol triphosphate. Taken together, these results suggest that protein kinase C function is indispensible for triggering L2C leukemic lymphocyte proliferation. The causes of this permanent activation merit further investigation. PMID- 2548043 TI - Specific binding of radioiodinated human GM-CSF to the blast cells of acute myeloblastic leukemia. AB - We describe here the presence of two classes of binding sites for GM-CSF expressed on blasts freshly isolated from five AML patients and one patient with CML in blastic phase: one of high-affinity (38-177 per cell, KD 8-150 pM) and one of low-affinity (121-806 per cell, KD 503-2683 pM). No correlation is observed between the receptor number, receptor affinity, and the growth stimulatory effect of GM-CSF on leukemic blast progenitors. Blasts from two cases showed no or negligible response to GM-CSF but expressed comparable numbers of receptors when compared with the numbers expressed by the sensitive blasts. Our data suggest that significant proliferative effects of GM-CSF can occur at low levels of high affinity receptor occupancy. Lack of responsiveness to GM-CSF in some AML patients is not correlated to the absence of GM-CSF receptors on leukemic cells. Reduction in the growth of blast progenitors at high concentrations of GM-CSF may be attributed to the differentiating activity of GM-CSF via low-affinity receptors on leukemic cells. PMID- 2548044 TI - Maturation induction in freshly isolated human myeloid leukemic cells, 1.25 (OH)2 vitamin D3 being the most potent inducer. AB - From ten consecutive patients with acute myeloid leukemia leukemic cells were isolated and cultured with and without 10(-6), 10(-7) and 10(-8) M 1.25(OH)2D3 (vit D3), retinoic acid (RA) cytosine-arabinoside (ARA-C) and 1.0, 1.25 and 1.5% dimethyl sulfoxide (DMSO). Maturation was measured with a comprehensive panel of qualitative and quantitative parameters of maturation. Six of those ten leukemias showed significant (p less than 0.01) changes in at least three parameters after exposure to either one of the differentiation inducers. Vit D3 induced maturation in four leukemias, in three of them clearly in monocytic direction. ARA-C showed changes in one leukemia in only three parameters not pointing to either granulocytic or monocytic direction. Maturation in granulocytic direction was observed after exposure to RA in one leukemia. Maturation induction was observed in six out of ten freshly isolated leukemic cells with vit D3 being the most potent inducer of maturation in monocytic direction. The data about inducibility of maturation in freshly isolated human leukemic cells are reviewed and discussed. PMID- 2548045 TI - Neurotoxicity of excitatory amino acid receptor agonists in young rat hippocampal slices. AB - Hippocampal slices from young (8-day-old) rats were evaluated as a model for investigating the mechanisms underlying the neurotoxic action of excitatory amino acid receptor agonists. The slices were exposed to the agonists for up to 30 min and were then postincubated for 90 min in order to allow irreversibly damaged cells to become visibly necrotic. Under control conditions (greater than or equal to 3 h incubation) all regions of the hippocampus and dentate gyrus displayed good preservation. Exposure of the slices to N-methyl-D-aspartate (NMDA) resulted in widespread, oedematous necrosis of all neuronal types (except undifferentiated granule cells) which was maximal after 20 min exposure to a concentration of 100 microM. With 30 min exposure, the EC50 for NMDA was 30 microM; 10 min exposure to NMDA at a concentration of 100 microM was sufficient to destroy 50% of the neurones. Quisqualate produced a degeneration of most (98%) of the CA3 neurones, a proportion (65%) of CA1 neurons and some (25%) of the dentate granule cells. The occurrence of "dark cell degeneration" was prevalent. Half maximal effects on CA3 neurones were estimated to be produced by a concentration of 15 microM (with 30 min exposure) or by 8 min exposure (at 100 microM concentration). Incubation of the slices with kainate (100 microM for 30 min) did not cause widespread damage but led to the necrosis of a small population of cells scattered in all regions of the hippocampus and dentate gyrus. The patterns of toxicity of the different agonists resemble closely those found after their administration in vivo. It is suggested that the hippocampal slices provide a valuable new model system for studying excitatory amino acid toxicity. PMID- 2548046 TI - Association of a mature B cell leukemia with a 4p+ chromosomal abnormality: derivation and characterization of a cell line. AB - We have found a single 4p+ chromosomal abnormality, 46,XX, -4, +der(4)t(3;4)(q13.3;p16), in a patient with an unusual B cell leukemia of mature phenotype characterized by a high white cell count, tartrate-resistant acid phosphatase-positive malignant cells, splenic white pulp proliferation, and a serum IgM monoclonal gammopathy. The malignant cells were characterized by surface expression of CD19 (B4), CD20 (B1), IgM, IgD, kappa, and HLA-DR. They were weakly positive for CD21 (B2) and negative for CD25 (interleukin-2 receptor). The malignant cells also showed clonal rearrangement of the immunoglobulin heavy chain and kappa light chain genes. A cell line, designated HCLW-3B, was derived from unstimulated peripheral blood obtained during the leukemic phase and was found to contain the same 4p+ chromosomal abnormality as well as genomic sequences of the Epstein-Barr virus nuclear antigen. A somatic cell hybrid constructed from HCLW-3B containing the derivative chromosome 4 was used to confirm that chromosome 3q was the source of the translocated material. The availability of a cell line which is clonally derived from the patient's circulating leukemia cells should permit further characterization of this translocation at the molecular level. PMID- 2548047 TI - [Effect of urea and uric acid on the hemolytic activity of Sendai virus]. AB - It was found that haemolytic activity of Fushimi strain of Sendai virus multiplied in allantoic cavity of chicken embryos is independent on its haemagglutinating titer and also on allantoic fluid urea and uric acid content. It was shown in experiments with embryonated eggs that these two compounds have no also influence on haemolytic activity induction in Sendai virus. Moreover, the results of an experiment in which allantoic fluid was replaced by Eagle's liquid suggest that most probably the other components present in allantoic fluid do not also influence the appearance of haemolytic activity of this virus. PMID- 2548048 TI - Activities of enzymes of collagen biosynthesis and levels of type III procollagen peptide in the serum of patients with sarcoidosis. AB - Serum immunoreactive prolylhydroxylase (IRPH), galactosylhydroxylsyl glucosyltransferase activity (GGT) and amino-terminal propeptides of type III procollagen (Pro(III) peptide) were measured in fifty three patients with sarcoidosis (all having some degrees of pulmonary fibrosis). The levels of IRPH and Pro(III) peptide showed no relationships to the clinical assessment of the disease and while GGT activity was raised in approximately 80% of the patients there was no correlation between the size of the increases and the clinical activity of the disease. The results of this study would suggest that measurement of the above parameters offer no specificity in either diagnosing or assessing the clinical activity of sarcoidosis. The observed increases in serum GGT activity in affected patients would however suggest that measurement of this enzyme may be useful perhaps in more severe pulmonary fibrotic reactions. PMID- 2548049 TI - Transverse relaxation rate enhancement caused by magnetic particulates. AB - Magnetic particulates have been shown to be powerful transverse relaxation enhancers and are under consideration as an MR contrast agent for the detection of liver and spleen lesions. This work describes the magnetic properties of a commercially available magnetic particulate and a Monte Carlo simulation of the effect of these particles on the transverse relaxation rates of water protons for spin-echo experiments. From the simultations, empirical relations were developed to describe the dependence of the enhancement of particle size, and concentration as well as the diffusion constant of water and the pulse spacing of a Carr Purcell-Meiboom-Gill pulse sequence used to measure the transverse relaxation time. The simulations are shown to agree with measurements of relaxation rates in agar samples containing the magnetic particulates. PMID- 2548050 TI - In vivo T1 characterization of genetically induced muscle atrophy. AB - In vivo spin-lattice relaxation times, T1, of water and lipid protons of normal and atrophic muscles were measured, using the spatially resolved spectroscopy (SPARS) sequence, in a genetic avian model of myopathy. These T1 values were compared with those of the hypertrophic muscles. Although the water T1 values of muscles were elevated in both types of lesions, the atrophic muscles showed a greater increase (54%) than the hypertrophic muscles (22%). The water T1 differentiation between the atrophic and hypertrophic muscles appeared to depend upon their bound water fractions that were calculated on the basis of the Fast Proton Diffusion model. The lipid T1 values of muscles were higher in the atrophic line of chickens compared to their genetic controls. In contrast, the lipid T1 values of muscles of the hypertrophic chickens and their controls were essentially identical. This suggests that the lipid T1 values may potentially complement the water T1 values in the differential diagnosis of muscle disorders. PMID- 2548052 TI - 1H-NMR relaxation times and water content of red blood cells from chronic alcoholic patients during withdrawal. AB - Red blood cell proton nmr, T1 and T2 times, and water content were measured for normal control subjects and thirteen patients with chronic alcoholism during the withdrawal phase. T1 and T2 were significantly increased without significant alteration in cell water content. The relaxation times were more markedly affected in those with symptoms of delirium tremens. These findings suggest that the intracellular free:bound water states rather than water content is the more likely explanation for these changes. The results are discussed in relation to similar findings obtained from in vivo studies in man and in vitro studies in animals. PMID- 2548051 TI - Magnetic resonance imaging of malignant fibrous histiocytoma. AB - The magnetic resonance imaging (MRI) changes in 39 patients with malignant fibrous histiocytoma (MFH) were reviewed retrospectively. Twenty-one sarcomas were in the lower extremity, five each in the upper extremity and trunk, two each in the neck and heart, and one each in the maxillary sinus, sella turcica, tongue, and spermatic cord. The examinations were performed with spin-echo sequences on a 1.5 Tesla Signa Scanner (GE, Milwaukee WI). Twenty-two tumors exhibited intermediate signal intensity on T1-weighted images and 23 were of high signal intensity on T2-weighted images. There was no significant differences in signal intensity of 12 preoperative and 13 recurrent neoplasms. Twelve of 13 patients were correctly diagnosed as having postoperative changes. The MR sensitivity and specificity for detecting a neoplasm were 96% and 83% respectively, but the signal changes were nonspecific for MFH. When compared to CT in 14 patients, MR better defined the extent of the MFH, its relationship to surrounding tissues and vessels, and best differentiated residual or recurrent disease from postoperative changes when examined at least 3 months after surgery. PMID- 2548053 TI - [The TESTASCAN single-photon emission multidetector computed gamma tomograph]. AB - A TESTASCAN single-photon emission multidetector gamma tomograph is a special purpose unit designed to investigate RP distribution in the brain. It permits obtaining transversal tomographic images with the resolution of about 10 mm, the thickness of an emitted layer being about 15 mm. The time of patient's examination during obtaining an image of 10-12 layers does not exceed 30 min. and can be reduced by increasing RP activity and minimizing the number of emitted transversal sections. The clinical testing of the unit has shown that it is reliable in maintenance and simple to operate during radionuclide data collection, reproduction and analysis. Emission computerized tomography in brain tumors can be regarded as a method increasing the accuracy and reliability of radionuclide diagnostic examination of neurosurgical and neurological patients. PMID- 2548054 TI - [The diagnostic importance of a quantitative evaluation of gamma scintigraphy in Perthes' disease in children]. AB - The investigation was performed in 28 children with Perthes disease, stages I III. The comparison of visual and quantitative assessment of gamma-scintigrams has shown that the use of the coefficient of differential accumulation as a criterion of asymmetry of RP accumulation in the hip joints makes it possible to increase 4-fold the effectiveness of diagnosis at stage I of Perthes disease, the most difficult one for x-ray diagnosis. At more advanced stages neither visual nor quantitative assessment of gamma-scintigrams is superior to roentgenography. Therefore the use of gamma-scintigraphy seems appropriate in Perthes disease at stage I. PMID- 2548055 TI - [Calcium-regulating hormones in endogenous hypercorticism]. AB - Investigation of 44 patients with endogenous hypercorticism (EH) of various degrees of severity showed that the development of osteoporosis was accompanied by changes in the indices of calcium-phosphorus metabolism and calcium regulating hormones. Marked variations in the level of parathyroidin, calcitonin, vitamin D3 were observed in a severe type of EH. All the examinees were characterized by a decrease in the transport form of vitamin D3, which was most noticeable in a mild form of EH. A significant decrease in the concentration of the transport form of vitamin D3 against a background of hypercalcemia and hypercalciuria in mild EH can be regarded as the most informative indicators in early diagnosis of initial symptoms of osteoporosis. PMID- 2548056 TI - High field localized proton spectroscopy in small volumes: greatly improved localization and shimming using shielded strong gradients. AB - The availability of shielded gradients facilitates accurate single-voxel spectroscopy. Exact localization (90-99%) of volumes as small as 2 x 2 x 2 mm is demonstrated using a modified stimulated echo sequence. Strong gradient pulses (up to 200 mT/m) are employed to clear the stimulated echo of unwanted magnetization, enabling superior (single-scan) shimming of the selected volume. In vivo and in vitro proton NMR results at 4.7 T are presented. PMID- 2548058 TI - Isolation, molecular characterization and expression of the ushB gene of Salmonella typhimurium which encodes a membrane-bound UDP-sugar hydrolase. AB - The UDP-sugar hydrolase of Salmonella typhimurium has previously been reported to be located in both the inner and the outer membrane. We have cloned the gene, designated ushB, encoding this enzyme and determined its nucleotide sequence. No significant sequence homology with the periplasmic UDP-sugar hydrolase of Escherichia coli was found at either the DNA or protein level. However, a sequence is detectable, in the E. coli genome, which weakly hybridizes with a specific ushB probe. Polypeptide analysis has allowed the identification of the Salmonella hydrolase which has an Mr of 28,349 as compared to an Mr of 60,767 for the E. coli hydrolase. Most of the protein (approximately 90%) is located in the inner membrane. Two independent membrane fractionation procedures indicate that the remainder may be associated with the outer membrane. The deduced primary structure indicates the presence of an N-terminal signal peptide, although certain features of the region surrounding the putative processing site indicate that processing may be inefficient, or may not occur. Experiments with several inhibitors of signal peptidase function fail to demonstrate the appearance of a precursor form. PMID- 2548057 TI - Trimethoprim resistance transposon Tn4003 from Staphylococcus aureus encodes genes for a dihydrofolate reductase and thymidylate synthetase flanked by three copies of IS257. AB - Trimethoprim resistance mediated by the Staphylococcus aureus multi-resistance plasmid pSK1 is encoded by a structure with characteristics of a composite transposon which we have designated Tn4003. Nucleotide sequence analysis of Tn4003 revealed it to be 4717 bp in length and to contain three copies of the insertion element IS257 (789-790 bp), the outside two of which are flanked by directly repeated 8-bp target sequences. IS257 has imperfect terminal inverted repeats of 27-28 bp and encodes for a putative transposase with two potential alpha-helix-turn-alpha-helix DNA recognition motifs. IS257 shares sequence similarities with members of the IS15 family of insertion sequences from Gram negative bacteria and with ISS1 from Streptococcus lactis. The central region of the transposon contains the dfrA gene that specifies the S1 dihydrofolate reductase (DHFR) responsible for trimethoprim resistance. The S1 enzyme shows sequence homology with type I and V trimethoprim-resistant DHFRs from Gram negative bacteria and with chromosomally encoded DHFRs from Gram-positive and Gram-negative bacteria. 5' to dfrA is a thymidylate synthetase gene, designated thyE. PMID- 2548059 TI - Mistranslation induces the heat-shock response in the yeast Saccharomyces cerevisiae. AB - The synthesis of heat-shock proteins can be triggered by a variety of stress inducing conditions. Here we show that translational misreading caused by growth in the presence of the aminoglycoside antibiotic paromomycin will induce the heat shock response in the yeast Saccharomyces cerevisiae. This was demonstrated (i) by the acquisition of thermotolerance, and (ii) by elevated levels of expression of the heat-shock protein, hsp70. In addition, transcription of the ubiquitin gene (UB14) was increased in paromomycin-grown cells. Control experiments with the protein synthesis inhibitor cycloheximide (which does not induce translational misreading) demonstrated that the response was not due to inhibition of protein synthesis per se. These observations strongly suggest that the synthesis of abnormally high levels of aberrant proteins is the trigger of the heat-shock response in this simple eukaryote. PMID- 2548060 TI - Nucleotide sequence and analysis of the N-terminal coding region of the Spodoptera-active delta-endotoxin gene of Bacillus thuringiensis aizawai 7.29. AB - The nucleotide sequence of a 2711bp DNA segment which contains the N-terminal coding sequence and the 5' flanking region of a crystal protein gene (bta) from Bacillus thuringiensis subsp. aizawai 7.29 has been determined. The coding region encodes an 824 amino-acid polypeptide corresponding to a carboxy-terminally truncated delta-endotoxin specifically active against the cotton leaf worm Spodoptera littoralis. Comparison of the deduced amino acid sequence of the bta gene with that of the 4.5, 5.3 and 6.6 kb classes of lepidopteran-active delta endotoxins revealed that the Bta sequence contains a very high level of amino acid substitutions in the N-terminal part of the protoxin molecule. The substitutions are grouped in several highly variable segments separated by highly conserved regions. These conserved domains are also present in the dipteran- and coleopteran-active delta-endotoxins. The control region of the bta gene shows considerable DNA identity with the control regions of the other lepidopteran active genes. Deletions of the 3' region of the gene were carried out and the toxic fraction of the bta delta-endotoxin was identified with the N-terminal half of the molecule. PMID- 2548061 TI - The Escherichia coli protein, Fis: specific binding to the ends of phage Mu DNA and modulation of phage growth. AB - We show, using gel retardation, that crude Escherichia coli cell extracts contain a protein which binds specifically to DNA fragments carrying either end of the phage Mu genome. We have identified this protein as Fis, a factor involved in several site-specific recombinational switches. Furthermore, we show that induction of a Mucts62 prophage in a fis lysogen occurs at a lower temperature than that of a wild-type strain, and that spontaneous induction of Mucts62 is increased in the fis mutant. DNasel footprinting using either crude extracts or purified Fis indicate that binding on the left end of Mu occurs at a site which overlaps a weak transposase binding site. Thus, Fis may modulate Mu growth by influencing the binding of transposase, or other proteins, to the transposase binding site(s), in a way similar to its influence on Xis binding in phage lambda. PMID- 2548062 TI - DNA supercoiling in Escherichia coli: topA mutations can be suppressed by DNA amplifications involving the tolC locus. AB - The level of DNA supercoiling is crucial for many cellular processes, including gene expression, and is determined, primarily, by the opposing actions of two enzymes: topoisomerase I and DNA gyrase. Escherichia coli strains lacking topoisomerase I (topA mutants) normally fail to grow in the absence of compensatory mutations which are presumed to relax DNA. We have found that, in media of low osmolarity, topA mutants are viable in the absence of any compensatory mutation, consistent with the view that decreased extracellular osmolarity causes a relaxation of cellular DNA. At higher osmolarity most compensatory mutations, as expected, are in the gyrA and gyrB genes. The only other locus at which compensatory mutations arise, designated toc, is shown to involve the amplification of a region of chromosomal DNA which includes the tolC gene. However, amplification of tolC alone is insufficient to explain the phenotypes of toc mutants. tolC insertion mutations alter the distribution of plasmid topoisomers in vivo. This effect is probably indirect, possibly a result of altered membrane structure and an alteration in the cell's osmotic barrier. As tolC is a highly pleiotropic locus, affecting the expression of many genes, it is possible that some of the TolC phenotypes are a direct result of this topological change. The possible relationship between toc and tolC mutations, and the means by which tolC mutations might affect DNA supercoiling, are discussed. PMID- 2548063 TI - Molecular characterization and sequence of phosphatidylinositol-specific phospholipase C of Bacillus thuringiensis. AB - The gene encoding monophosphatidylinositol inositol phosphohydrolase (PI-specific phospholipase C, PI-PLC) of Bacillus thuringiensis was cloned in Staphylococcus carnosus TM300. The complete coding region comprises 987 base pairs corresponding to a precursor protein of 329 amino acids (molecular weight, 38,095). The NH2 terminal sequence of the purified enzyme from Escherichia coli indicated that the mature PI-PLC consists of 299 amino acid residues with a molecular weight of 34,586. Polyacrylamide gel electrophoresis revealed the same molecular weight for the purified enzyme isolated from the DNA-donor strain of B. thuringiensis and from the E. coli clone. By computer analysis, the secondary structure was predicted. The enzyme from the E. coli recombinant shows no activity on other phospholipids and sphingomyelin. The cleaving specificity of PI-PLC was examined by thin layer chromatography. PMID- 2548064 TI - A bacterial c-type cytochrome can be translocated to the periplasm as an apo form; the biosynthesis of cytochrome cd1 (nitrite reductase) from Paracoccus denitrificans. AB - An apo form of cytochrome cd1 (nitrite reductase) of Paracoccus denitrificans has been detected immunologically in the periplasm of a mutant that lacks all c-type cytochromes. A method for the preparation of apo-nitrite reductase (lacking both c- and d-type haem) from the holoenzyme of wild-type cells has been developed. The apoprotein synthesized by the mutant is indistinguishable from the chemically prepared apoprotein in respect of: (i) subunit molecular weight; (ii) formation of a homodimer; (iii) properties on anion exchange chromatography. The holoenzyme has similar properties in respect of (i) and (ii) but behaves differently during anion exchange. A suggested mode of assembly of cytochrome cd1 is translocation into the periplasm of a precursor polypeptide, maturation by a signal peptidase to give an apoprotein identical to that prepared chemically from the holoenzyme, followed by insertion of c-type and d-type haem in an as yet unknown order. PMID- 2548065 TI - 86Rb(K) influx and [3H]ouabain binding by human platelets: evidence for beta adrenergic stimulation of Na-K ATPase activity. AB - Although active transport of potassium into human platelets has been demonstrated previously, there is hitherto no evidence that human platelets have an ouabain inhibitable Na-K ATPase in their membrane. The present study demonstrates active rubidium (used as an index of potassium influx), 86Rb(K), influx into platelets, inhibitable by ouabain, and also demonstrates the presence of specific [3H]ouabain binding by the human platelet. This 86Rb(K) influx was stimulated by adrenaline, isoprenaline, and salbutamol, but noradrenaline caused a mild inhibition. Active 86Rb(K) influx by platelets was inhibited markedly by timolol, mildly by atenolol, but not by phentolamine. Therefore, active 86Rb(K) influx in human platelets is enhanced by stimulation of beta adrenoceptors of the beta 2 subtype. The platelet may therefore replace the leukocyte in future studies of Na K ATPase activity. This would be a considerable advantage in view of the ease and rapidity of preparation of platelets. PMID- 2548067 TI - Cerebral glutamine metabolism: study of modulatory effects of glutamine on gamma aminobutyric acid-ergic neurotransmission. AB - Glutamine is one major precursor of gamma-aminobutyric acid (GABA) and glutamate, the most important inhibitory and excitatory neurotransmitters in the mammalian brain, respectively. Changes in cerebral glutamine concentrations occur in various metabolic encephalopathies including hyperammonemia and liver failure. As glutamine inhibits the specific binding of GABA to its postsynaptic receptor at physiologic concentrations, in this study the effects of glutamine on various components of the GABAA-benzodiazepine receptor complex were studied. Glutamine dose dependently inhibited the stimulation of flunitrazepam binding by GABA. This inhibition occurred already at concentrations of 10 mumol/L glutamine. Glutamine had no effects on basal or GABA-stimulated synaptoneurosomal chloride uptake. It is concluded that glutamine is not a modulator of the GABAA-benzodiazepine neurotransmitter system. Thus, changes of cerebral glutamine concentrations are unlikely to contribute to the activation of GABA-ergic neurotransmission in liver failure. PMID- 2548066 TI - Galactose as a regulatory factor of its own metabolism by rat liver. AB - As part of a series of studies to assess the regulation of hepatic galactose metabolizing enzymes, galactokinase, galactose-1-phosphate uridyltransferase, and UDPgalactose-4-epimerase, the effect of feeding a high galactose-containing diet to normal adult and pregnant female rats was examined. Sixteen days of galactose exposure of adult virgin females produced a different response in the specific activity of each of the enzymes, that of galactokinase being lower, transferase higher, and epimerase transiently elevated. Galactose feeding increased the specific activity of transferase in pregnant rat liver above the elevated level that already exists in the pregnant state but failed to influence the enzyme of the developing fetal liver. Galactose added to liver homogenates did not activate transferase. The increased activity in liver of adult, fed animals was not associated with a change in isoenzyme patterns examined by isoelectric gel electrophoresis but was characterized on kinetic analysis by an increase in Vmax for UDPglucose. The changes in enzyme specific activity in liver of animals fed galactose appear to have physiologic significance because hepatocytes isolated from galactose exposed livers take up more galactose and convert more to glucose and lactate than cells from control animals. PMID- 2548068 TI - [Enzymes involved in the metabolism of flavin nucleotides in Streptomyces olivaceus]. AB - The work was aimed at studying enzymes involved in the metabolism of flavin nucleotides, namely, riboflavin kinase (EC 2.7.1.26) and FAD pyrophosphorylase (EC 2.7.7.2), as well as flavin mononucleotide hydrolysis by acid phosphatase (EC 3.1.3.2) and alkaline phosphatase (EC 3.1.3.1) in Streptomyces olivaceus actively producing vitamin B12. No correlation could be established between changes in the activity of the above enzymes during the culture growth and the qualitative composition of flavins. The enzyme activity was assayed using, as an enzyme preparation, both intact cells and a cell-free extract obtained by disintegrating the mycelium with different techniques. The screening effect of phosphatases exerted when the activity of riboflavin kinase was assayed could be partly eliminated by adding sodium fluoride to the incubation medium. The localisation of the above enzymes in the cytoplasm is discussed. PMID- 2548069 TI - Fatal rhabdomyolysis and renal failure associated with hand, foot and mouth disease. AB - Fatal severe rhabdomyolysis and anuric renal failure developed in a young man who had contracted hand, foot and mouth disease during an epidemic of this disease. Viral studies implicated coxsackievirus A16 as the infecting agent. This appears to be the first reported case of rhabdomyolysis that was associated with hand, foot and mouth disease and infection with coxsackievirus A16. PMID- 2548071 TI - Effects of systemic alkalosis on urinary magnesium excretion in the rat. AB - Metabolic alkalosis has previously been shown to have an antimagnesiuric influence. To further clarify this phenomenon, short-term clearance studies were performed on intact anesthetized rats subjected to 0.9% saline infusion, 0.15 M NaHCO3 infusion or acute respiratory alkalosis. The experimental protocols resulted in a similar degree of natriuresis in each of the three groups. The increase in plasma pH value was similar both in animals treated with NaHCO3 and animals with respiratory alkalosis. Filtered loads of Mg did not differ in the three experimental groups. However, only acute metabolic alkalosis was associated with a reduction in the absolute rate of Mg excretion (saline: 0.49 +/- 0.05 mu Eq/min; 0.15 M NaHCO3: 0.29 +/- 0.04 mu Eq/min; acute respiratory alkalosis: 0.48 +/- 0.03 mu Eq/min) and fractional Mg excretion (saline: 40.3 +/- 5.3%; 0.15 NaHCO3: 18.7 +/- 1.4%; acute respiratory alkalosis: 37.2 +/- 6.9%). A similar decrease in urinary Mg excretion in animals treated with bicarbonate infusion was observed following removal of the parathyroid gland. Moreover, for any given rate of urinary Na excretion, Mg excretion was lower in bicarbonate-treated animals than in rats infused with saline solution. Intact animals treated with increasing doses of NaHCO3 revealed a statistically significant inverse correlation between the Mg to Na clearance ratio and urinary and plasma bicarbonate concentration. In contrast, such a correlation was not observed during respiratory alkalosis. The findings suggest that bicarbonate ion directly stimulates tubular magnesium reabsorption independent of the presence or absence of parathyroid hormone. PMID- 2548070 TI - Effect of hypocalcemia on renal phosphorus handling in the rat: interaction with dietary phosphorus and PTH. AB - The effect of Na-EGTA induced hypocalcemia was investigated in chronically parathyroidectomized (PTX) rats. In dietary inorganic phosphorus (Pi) replete animals, reduction in the plasma calcium to 1.37 +/- 0.03 mM at constant filtered loads of Pi had no effect on tubular reabsorption of phosphorus (85 +/- 3 vs. 83 +/- 2 micrograms/ml, NS). In dietary Pi-deprived rats, where an elevation of plasma calcium is a known accompaniment, similar reduction in plasma calcium concentration also failed to alter (89 +/- 2 vs. 90 +/- 2 micrograms/ml, NS) phosphorus reabsorption. Resistance to parathyroid hormone (PTH) in dietary Pi deprivation was confirmed. Super imposition of PTH on EGTA induced hypocalcemia during dietary Pi deprivation, however, resulted in a significant reduction in tubular Pi reabsorption (81 +/- 2 vs. 67 +/- 3 micrograms/ml, p less than 0.05) with full restoration of its phosphaturic action. These changes were unaccompanied by differences in urinary adenosine 3',5'-cyclic phosphate (cAMP). In conclusion, hypocalcemia per se does not alter the renal handling of phosphorus at constant plasma Pi concentrations. The elevation in the plasma calcium associated with dietary Pi deprivation does not contribute importantly to the hypophosphaturia of dietary Pi deprivation. Hypocalcemia, however, abolishes the resistance to PTH observed in Pi deprivation. PMID- 2548072 TI - Aminoaciduria of vitamin D deficiency is independent of PTH levels and urinary cyclic AMP. AB - Aminoaciduria and secondary hyperparathyroidism accompany vitamin D deficiency. However, the degree of aminoaciduria and PTH elevation have not been studied relative to different calcium and phosphorus dietary intakes. Weanling rats were fed 5 vitamin D deficient diets for 4-6 weeks: very low Ca (VLC) 0.02% Ca, 0.3% P; VLC + 1,25-dihydroxyvitamin D [1,25(OH)2D3], same + 500 pmol i.p. for 2 days; low Ca (LC) 0.45% Ca, 0.3% P; very low P (VLP) 1.2% Ca, 0.1% P; high Ca (HC) 2.5% Ca, 0.3% P, and control 1.2% Ca, 0.70% P + 2.5 micrograms% vitamin D. Amino acids, serum 25-hydroxyvitamin D [25(OH)D3], 1,25(OH)2D3, and PTH, using a specific antirat PTH antibody, were measured. A significant generalized aminoaciduria (11 amino acids) was found in all vitamin D-deficient groups. Furthermore, it was independent of plasma Ca and PTH, and urinary cAMP excretion irrespective of diet. Serum 25(OH)D and 1,25(OH)2D were significantly reduced in all vitamin D-deficient groups. VLC and VLC + 1,25(OH)2D3 were associated with the highest PTH levels (10- and 13-fold increase, respectively) and urinary cAMP (2.3-fold increase in each) and the lowest serum Ca. LC rats had an 8.8- and a 1.7-fold increase in PTH and urinary cAMP, respectively. Phosphate depletion was found in VLP rats documented by insignificantly elevated PTH, normal urinary cAMP, hypercalciuria, and percent tubular reabsorption of phosphate of greater than 99%. While dietary Ca and P affect plasma and urinary Ca and P plasma PTH and urinary cAMP, it appears that dietary P affects the aminoaciduria observed in this study via mechanisms that remain unclear. The possibility that the mechanism for the tubulopathy is multifactorial should be entertained. PMID- 2548073 TI - Cushing's disease and prolonged spontaneous remission. Clinical case. AB - We describe a case of prolonged spontaneous remission of Cushing's disease lasting since 1986 and documented both clinically and biochemically. This case confirms previous sporadic observations based mainly on clinical findings and indicates that spontaneous remission of Cushing's disease, although rare, may exist. The Authors outline that in some cases of Cushing's disease it could be convenient to adopt a conservative policy with frequent outpatient reviews in order to observe the natural course of the disease, particularly when the clinical findings are mild and the original lesion is not recognized with the available technology. PMID- 2548074 TI - Enterovirus surveillance--United States, 1989. PMID- 2548075 TI - [Measurement of portal blood flow in man by a continuous local thermodilution method. III. Effects of dopamine on systemic and portal hemodynamics after hepatectomy]. AB - We gave dopamine (3 micrograms/kg/min, 30 min) intravenously on the third day after hepatectomy to 19 patients, and studied the effects of the drug on systemic and portal hemodynamics. In another 42 patients, administration of dopamine at the same rate was started soon after hepatectomy and continued for about 2 weeks; the clinical results were evaluated. After hepatectomy, the systemic hemodynamics were hyperdynamic and the portal hemodynamics were hypodynamic. After 30 min of dopamine administration, the oxygen pressure in portal blood increased, and because portal blood flow also increased, the oxygen delivery to the liver increased. The mechanism involved an increased proportion of portal venous flow to cardiac output, and a decrease in the splanchnic resistance, not portal venous resistance. Probably, specific dopamine receptors played important role in the increase in the superior mesenteric arterial blood flow. Among 42 patients given small dosage of dopamine, the clinical symptoms of five of seven who had developed liver failure improved. None of the other 35 patients given dopamine preventively developed liver failure. Dopamine in small doses is useful for the management of liver failure after liver resection. PMID- 2548076 TI - GPA1Val-50 mutation in the mating-factor signaling pathway in Saccharomyces cerevisiae. AB - The GPA1 gene of Saccharomyces cerevisiae encodes a protein that is highly homologous to the alpha subunit of mammalian hetrotrimeric G proteins and is essential for haploid cell growth. A mutation of the GPA1 protein, GPA1Val-50, in which Gly-50 was replaced by valine, could complement the growth defect of a GPA1 disruption, gpal::HIS3. However, cells with gpa1::HIS3 expressing the GPA1Val-50 protein were supersensitive to alpha-factor in a short-term incubation but resumed growth after long-term incubation even after exposure to high concentrations of alpha-factor. The former phenotype associated with GPA1Val-50 is recessive, and the latter phenotype is dominant to GPA1+. The supersensitivity of GPA1Val-50 to alpha-factor was dependent on STE2 and STE4, which demonstrates that this GPA1Val-50-produced phenotype requires the mating-factor receptor and the beta subunit of the G protein. The double mutant of sst2-1 GPA1Val-50 recovered from division arrest, which suggested that SST2 is not required for recovery of the GPA1Val-50 mutant. PMID- 2548077 TI - Involvement of second messengers in regulation of the low-density lipoprotein receptor gene. AB - Transcription of the low-density lipoprotein receptor (LDL-R) gene in the human monocytic leukemic cell line THP-1 and in the human hepatocarcinoma cell line Hep G2 is regulated by second messengers of the diacylglycerol-protein kinase C (DAG PKC), inositol 1,4,5-triphosphate-Ca2+, and cyclic AMP pathways. Exogenous phospholipase C (which releases DAG and inositol 1,4,5-triphosphate), PKC activators (phorbol esters and DAG), Ca2+ ionophores, and a cyclic AMP analog all transiently induced accumulation of LDL-R mRNA. The effects of these three signal transducing pathways were to a large extent additive. Furthermore, PKC stimulation effected an increase in LDL binding, which suggested that the increase in LDL-R mRNA resulted in an increase in functional cell surface receptor activity. These results suggest that uptake of cholesterol by these cells is under control of both intracellular cholesterol levels and external signals. PMID- 2548079 TI - An embryonic enhancer determines the temporal activation of a sea urchin late H1 gene. AB - Normal development requires that individual genes be expressed in their correct temporal patterns, but the mechanisms regulating this process during early embryogenesis are poorly understood. We have studied the early and late sea urchin histone genes during embryogenesis to address the molecular mechanisms controlling temporal gene expression. By measuring the changes in expression of cloned H1-beta DNA constructs after microinjection into fertilized one-cell zygotes, we demonstrated that a highly conserved 30-base-pair segment of DNA between positions -288 and -317 (USE IV) is responsible for the transcriptional activation of this late histone gene at the late blastula stage. In this report, we demonstrate that an oligonucleotide corresponding to USE IV acts as an embryonic enhancer element capable of activating the simian virus 40 early promoter in a stage-specific manner. Using an in vivo competition assay and in vitro DNase I footprinting and mobility shift assays, we also identified a protein(s) that interacts with this enhancer. Results of the competition assay suggested that this factor acts to stimulate transcription of the H1-beta gene. The factor was found to be stored in mature eggs as well as in all embryonic stages examined. The mobility of the factor found in eggs, however, differed from that of the embryonic form, which suggested that posttranslational modification occurs after fertilization. PMID- 2548078 TI - Use of a glucocorticoid-inducible promoter for expression of herpes simplex virus type 1 glycoprotein gC1, a cytotoxic protein in mammalian cells. AB - Abundant expression of herpes simplex virus type 1 glycoprotein gC (gC1) in transfected mammalian cells has not previously been achieved, possibly because gC1 protein is toxic to cells. To approach this problem, the gC1 coding sequence was placed under the control of the weak but inducible glucocorticoid-responsive promoter from the mouse mammary tumor virus (MMTV) long terminal repeat (LTR). As controls to evaluate for gC1 cytotoxicity, the MMTV LTR promoter was used to express glycoprotein gD1, and a strong, constitutive promoter from the Moloney murine sarcoma virus LTR was used to express gC1. L cells were transfected with these constructs, and a clone expressing gC1 from the inducible MMTV LTR promoter was analyzed. In the absence of glucocorticoid (dexamethasone) stimulation, only a low level of gC1 mRNA expression was detected; after overnight stimulation with dexamethasone, transcription increased approximately 200-fold. Abundant gC1 protein that was functionally active in that it bound complement component C3b, was produced. From passages 5 through 26 (70 cell population doublings), the gC1 producing clone became less responsive to overnight dexamethasone stimulation. The block to gC1 expression occurred at the level of transcription and was associated with hypermethylation of the MMTV LTR DNA. Treatment of the clone with 5-aza-2'-deoxycytidine partially reversed the block in gC1 protein production. Late-passage cells assumed a gC1-negative phenotype that appeared to offer a selective growth advantage, which suggested that gC1 was cytotoxic. Several findings support this view: (i) some cells expressing gC1 after overnight stimulation with dexamethasone assumed bizarre, syncytial shapes; (ii) continuous stimulation with dexamethasone for 5 weeks resulted in death of most cells; (iii) cells transfected with gC1 under the control of the strong Moloney murine sarcoma virus promoter assumed bizarre shapes, and stable gC1-expressing clones could not be established; and (iv) cells induced to express gD1 retained a normal appearance after overnight stimulation or 15 weeks of continuous stimulation with dexamethasone. The inducible MMTV LTR promoter is useful for expressing gC1 and may have applications for expressing other cytotoxic proteins. PMID- 2548080 TI - Regulation of mouse CYP1A1 gene expression by dioxin: requirement of two cis acting elements during induction. AB - The mouse cytochrome P1450 (CYP1A1) gene is responsible for the metabolism of numerous carcinogens and toxic chemicals. Induction by the environmental contaminant tetrachlorodibenzo-p-dioxin (TCDD) requires a functional aromatic hydrocarbon (Ah) receptor. We examined the 5'-flanking region of the CYP1A1 gene in mouse hepatoma Hepa-1 wild-type cells and a mutant line having a defect in chromatin binding of the TCDD-receptor complex. We identified two cis-acting elements (distal, -1071 to -901 region; proximal, -245 to -50 region) required for constitutive and TCDD-inducible CYP1A1 gene expression. Three classes of DNA protein complexes binding to the distal element were identified: class I, found only in the presence of TCDD and a functional Ah receptor, that was heat labile and not competed against by simian virus 40 (SV40) early promoter DNA; class II, consisting of at least three constitutive complexes that were heat stable and bound to SV40 DNA; and class III, composed of at least three constitutive complexes that were thermolabile and were not competed against by SV40 DNA. Essential contacts for these proteins were centered at -993 to -990 for the class I complex, -987, -986, or both for the class II complexes, and -938 to -927 for the class III complexes. The proximal element was absolutely essential for both constitutive and TCDD-inducible CYP1A1 gene expression, and at least two constitutive complexes bound to this region. These data are consistent with the proximal element that binds proteins being necessary but not sufficient for inducible gene expression; interaction of these proteins with those at the distal element was found to be required for full CYP1A1 induction by TCDD. PMID- 2548081 TI - In vitro activation and nuclear translocation of NF-kappa B catalyzed by cyclic AMP-dependent protein kinase and protein kinase C. AB - We have examined whether a precursor form of NF-kappa B, a DNA-binding protein that plays a role in the transcriptional control of several genes, including kappa immunoglobulin light chain and interleukin-2 receptor alpha subunit, could be activated in vitro by protein kinases. DNA-binding activity of NF-kappa B was induced in the cytosolic fraction of unstimulated 70Z/3 murine pre-B cells by incubation with the catalytic subunit of cyclic AMP-dependent protein kinase (PKA) or protein kinase C (PKC). In contrast, PKA and PKC did not activate NF kappa B in nuclear extracts from unstimulated cells. Identical results were obtained with the human natural killer-like cell line YT, which can be induced to express the interleukin-2 receptor alpha subunit in response to interleukin-1, cyclic AMP, or phorbol 12-myristate 13-acetate. Furthermore, when nuclei from unstimulated cells were incubated with PKA- or PKC-treated cytosolic fraction for 30 min at 30 degrees C, NF-kappa B was translocated into the nuclei. This translocation did not occur at 4 degrees C and was inhibited by wheat germ agglutinin but not by concanavalin A. Our findings support the conclusion that NF kappa B exists in the cytoplasm of unstimulated cells in an inactive form that can be converted by exposure to PKA or PKC to an active DNA-binding form that can translocate to the nucleus. PMID- 2548082 TI - Some of the signals for 3'-end formation in transcription of the Saccharomyces cerevisiae Ty-D15 element are immediately downstream of the initiation site. AB - Fragments from the Ty-D15 element of Saccharomyces cerevisiae were assayed for the ability to direct 3'-end formation for RNA initiated by the GAL1 promoter. The delta, the direct repeat at each end of the element, was capable of forming 3' ends at two sites, an inefficient upstream site and an efficient downstream site near the end of the delta. Different sequences were required for 3'-end formation at these sites. For the efficient site, all transcripts had 3' ends in the delta and no downstream transcription was detected, which suggested that these sequences terminate transcription. Surprisingly, the delta region downstream of the initiation site for Ty RNA comprised part of this major site and terminated more than 50% of the transcripts that read into it. Sequences necessary for the efficient site were localized to two small regions. Both regions were upstream of the 3' end and contained similarities to a tripartite consensus sequence that has been proposed as a terminator element. Sequences near the position of the 3' end could also affect termination; a short G + C-rich sequence inserted just downstream changed an efficient terminator to an inefficient one. Initiation in the delta had no effect on the efficiency or positions or termination in that delta. A new initiation site was seen when the same delta terminated transcription, but transcriptional interference did not occur, since the amount of initiation was not decreased. PMID- 2548083 TI - Molecular analysis of N6-methyladenine patterns in Tetrahymena thermophila nuclear DNA. AB - We have cloned two DNA fragments containing 5'-GATC-3' sites at which the adenine is methylated in the macronucleus of the ciliate Tetrahymena thermophila. Using these cloned fragments as molecular probes, we analyzed the maintenance of methylation patterns at two partially and two uniformly methylated sites. Our results suggest that a semiconservative copying model for maintenance of methylation is not sufficient to account for the methylation patterns we found during somatic growth of Tetrahymena. Although we detected hemimethylated molecules in macronuclear DNA, they were present in both replicating and nonreplicating DNA. In addition, we observed that a complex methylation pattern including partially methylated sites was maintained during vegetative growth. This required the activity of a methylase capable of recognizing and modifying sites specified by something other than hemimethylation. We suggest that a eucaryotic maintenance methylase may be capable of discriminating between potential methylation sites to ensure the inheritance of methylation patterns. PMID- 2548084 TI - Multiple proto-oncogene activations in avian leukosis virus-induced lymphomas: evidence for stage-specific events. AB - We have examined avian leukosis virus-induced B-cell lymphomas for multiple, stage-specific oncogene activations. Three targets for viral integration were identified: c-myb, c-myc, and a newly identified locus termed c-bic. The c-myb and c-myc genes were associated with different lymphoma phenotypes. The c-bic locus was a target for integration in one class of lymphomas, usually in conjunction with c-myc activation. The data indicate that c-myc and c-bic may act synergistically during lymphomagenesis and that c-bic is involved in late stages of tumor progression. PMID- 2548086 TI - Cell size modulation by CDC25 and RAS2 genes in Saccharomyces cerevisiae. AB - A detailed kinetic analysis of the cell cycle of cdc25-1, RAS2Val-19, or cdc25 1/RAS2Val-19 mutants during exponential growth is presented. At the permissive temperature (24 degrees C), cdc25-1 cells show a longer G1/unbudded phase of the cell cycle and have a smaller critical cell size required for budding without changing the growth rate in comparison to an isogenic wild type. The RAS2Val-19 mutation efficiently suppresses the ts growth defect of the cdc25-1 mutant at 36 degrees C and the increase of G1 phase at 24 degrees C. Moreover, it causes a marked increase of the critical cell mass required to enter into a new cell division cycle compared with that of the wild type. Since the critical cell mass is physiologically modulated by nutritional conditions, we have also studied the behavior of these mutants in different media. The increase in cell size caused by the RAS2Val-19 mutation is evident in all tested growth conditions, while the effect of cdc25-1 is apparently more pronounced in rich culture media. CDC25 and RAS2 gene products have been showed to control cell growth by regulating the cyclic AMP metabolic pathway. Experimental evidence reported herein suggests that the modulation of the critical cell size by CDC25 and RAS2 may involve adenylate cyclase. PMID- 2548085 TI - Yeast pheromone response pathway: characterization of a suppressor that restores mating to receptorless mutants. AB - Saccharomyces cerevisiae haploid cells, alpha and a, mate after being appropriately stimulated by the pheromone secreted by the opposite cell type (a factor and alpha-factor, respectively). The binding of a pheromone to its receptor is a signal that initiates a series of intracellular changes that lead to the specific physiological alterations required for mating. To identify components of the signal transduction pathway, we sought pseudorevertants that restored mating competence to receptor mutants (MAT alpha ste3::LEU2). The suppressor srm1-1 was isolated as a recessive mutation that conferred temperature sensitive growth to all strains and mating ability to MAT alpha ste3::LEU2 strains at the nonpermissive temperature. In addition, when srm1-1 mutants were shifted to the nonpermissive temperature, they exhibited two phenotypes characteristic of pheromone response, induction of FUS1 transcription and accumulation of cells in the G1 phase of the cell cycle. The srm1-1 mutation also suppressed a deletion of the alpha-factor-receptor gene in a cells. Together, these phenotypes suggest that the wild-type SRM1 product is a component of the pheromone response pathway. Deletion of STE4 or STE5, which are required in both haploid cell types for mating and response to pheromone, was not suppressed by srm1-1, suggesting that the SRM1 product may function before the STE4 and STE5 products. SRM1 is an essential gene and is expressed in both haploid cell types as well as in the product of their mating, a/alpha diploids. Homozygous srm1-1 a/alpha diploids were temperature sensitive although they did not arrest in G1. Thus, the SRM1 product may also have a role in the vegetative life cycle of cells. PMID- 2548087 TI - Regulation of the visna virus long terminal repeat in macrophages involves cellular factors that bind sequences containing AP-1 sites. AB - Visna virus gene expression is highly restricted in monocytes but is induced by monocyte-macrophage differentiation in vivo. Deletion and linker-scanning mutants, gel shift assays, and DNase I footprinting were used to identify sequences in the visna virus long terminal repeat involved in the developmental regulation of gene expression in the U937 monocytic cell line. We found that an AP-1 and an AP-4 binding site were critical for basal activity and that the AP-1 site was required for phorbol ester-inducible gene expression. These results suggest that cellular factors that interact with AP-1 sites are involved in the developmental regulation of visna virus gene expression in macrophages. PMID- 2548088 TI - High-resolution footprints of the DNA-binding domain of Epstein-Barr virus nuclear antigen 1. AB - The DNA-binding domain of Epstein-Barr virus nuclear antigen 1 was found by hydroxyl radical footprinting to protect backbone positions on one side of its DNA-binding site. The guanines contacted in the major groove by the DNA-binding domain of Epstein-Barr virus nuclear antigen 1 were identified by methylation protection. No difference was found in the interaction of the DNA-binding domain of Epstein-Barr virus nuclear antigen 1 with tandemly repeated and overlapping binding sites. PMID- 2548090 TI - Effect of cisplatin, lipopolysaccharide, muramyl dipeptide and recombinant interferon-gamma on murine macrophages in vitro. II. Production of H2O2, O2- and interleukin-1. AB - Murine macrophage monolayers treated with cisplatin, lipopolysaccharide (LPS), muramyl dipeptide (MDP) or recombinant interferon-gamma (rIFN gamma) for 2-48 h showed significant increases in the release of H2O2, O2- and interleukin-1 (IL-1) as compared to untreated macrophages. The treatment of macrophages with different combinations of the above agents did not induce synergistic or additive effects on the production of H2O2, O2- and IL-1. The priming of macrophages with rIFN gamma had a significant effect in the additional increased production of H2O2, O2 and IL-1 when subsequently treated with cisplatin, LPS or MDP. PMID- 2548091 TI - Temperature-dependent mutational specificity of an Escherichia coli mutator, dnaQ49, defective in 3'----5' exonuclease (proofreading) activity. AB - Escherichia coli strains carrying the temperature-dependent dnaQ49 allele are strong mutators at 37 degrees C. Since the dnaQ49 gene encodes the epsilon subunit of DNA polymerase III, it is thought that the large number of errors results in part from impaired proofreading activity during DNA replication. We have examined dnaQ49-induced reversion patterns of defined trpA alleles to determine the kinds of errors produced by dnaQ49 at 30 degrees C and 37 degrees C. We found that at 37 degrees C dnaQ49 produced all types of base-pair substitutions in addition to frameshifts with transitions generally occurring more frequently than transversions. This generalized mutator activity is very similar to that displayed in rich medium by mutD5, another mutator allele at the dnaQ locus. However, when dnaQ49 strains were cultured at 30 degrees C, not only were reversion frequencies much lower than at 37 degrees C, but in addition, the spectrum was altered. Transversions became proportionally more prevalent in the reversion spectra at the lower temperature. We suggest the possibility that at 37 degrees C dnaQ49 results in defective proofreading and methyl-directed postreplicative mismatch repair, while at 30 degrees C mismatch repair is fully and proofreading partially restored. PMID- 2548092 TI - Induction of chromosomal aberrations in CHO cells by restriction endonucleases: effects of blunt- and cohesive-ended double-strand breaks in cells treated by 'pellet' methods. AB - In recent reports it has been suggested that restriction endonucleases (RE) producing cohesive-ended double-strand breaks (dsb), are of comparable effectiveness to those producing blunt-ended dsb in causing chromosomal aberrations (CA) in mammalian cells. In several of these reports, trypsinized cells or suspension cultures were treated as cell 'pellets' in small volumes containing RE and storage buffers. In this study we have examined this by comparing 2 'pellet' methods in which trypsinized Chinese hamster cells were treated with RE in small volumes, after cells were centrifuged to a pellet. In the first method, cells were treated with RE in storage buffer as previously reported (e.g. Obe et al., 1985). In the second method, cells were treated as pellets with Sendai virus and purified RE. For both methods we show that the frequency of chromosomal aberrations was higher in cells treated with RE causing blunt-ended dsb than those causing cohesive-ended dsb. The first method however was found to lead to substantial loss in cell viability. The results strengthen the conclusion drawn from our earlier work, using treatment of attached V79 or CHO-K1 cells with Sendai virus, that cohesive-ended dsb are less effective than blunt-ended dsb in causing chromosomal aberrations. PMID- 2548089 TI - Splicing of a yeast intron containing an unusual 5' junction sequence. AB - The Saccharomyces cerevisiae COX5b gene contains a small intron that is unique in two respects. First, it interrupts the ATG codon that initiates translation of the COX5b product. Second, it contains a sequence at the 5' splice junction (5' GCATGT-3') that differs from the highly conserved yeast hexanucleotide (5' GTAPyGT-3') and from the 5'-GT found at the corresponding position in nearly all introns of eucaryotic protein-coding genes. We have analyzed both the transcripts derived from the COX5b gene and the splicing of its intron. We show here that an unspliced mRNA precursor constituted a minor fraction of the total COX5b message, even when the gene was overexpressed. We also show that both major transcripts derived from COX5b had been spliced. Our results suggest that at least in the case of COX5b, a 5'-GC can function as efficiently as the highly conserved 5'-GT in the splicing reaction. PMID- 2548093 TI - Mutational specificity studies of endogenous mammalian cell loci: methodological aspects. AB - We report here the details of a modified cloning method first described by Seed et al. (1983) for use in an extensive study of mutational specificity at the aprt locus of Chinese hamster ovary cells. The technology depends on homologous recombination between a suppressor probe plasmid and the desired insert in a genomic library constructed in a double amber mutant bacteriophage lambda vector. Recombinant phage form blue plaques on a non-suppressor, lacZ amber host. We have determined the sensitivity of the method. Homologous recombination frequency is in the order of 10(-3), 6-7 orders of magnitude above non-homologous events. Recombination efficiency is unaffected when the target phage is diluted 100,000 fold with parental vector. A background of plaques is observed along with the desired blue plaques. Although the color discrimination permits us to easily avoid these background plaques, we have characterized the frequency and the nature of these events. PMID- 2548094 TI - Identification of a multispecific lipoprotein receptor in adult Schistosoma japonicum by ligand blotting analyses. AB - A 43-kDa putative lipoprotein receptor (Sj43) of adult Schistosoma japonicum worms has been identified using ligand blotting techniques. Single and two dimensional electrophoretic analyses showed that Sj43 consisted of a single acidic polypeptide with multiple lipoprotein specificity. The molecule bound 125I labelled low-density (apo-B), very low-density or high-density (apo-A and/or apo C) lipoproteins from different mammalian hosts that are permissive to S. japonicum infection, but did not bind mouse apo-A containing lipoprotein. The binding of 125I-labelled lipoprotein to Sj43 could be inhibited by unlabelled human LDL, EDTA or Suramin, or by chemical modification of lipoprotein lysine or arginine residues. Sj43 was localised at the parasite's tegument and gut lining. PMID- 2548095 TI - Specificity spillover at the hormone receptor. PMID- 2548096 TI - Human parvovirus B19 infection among hospital staff members after contact with infected patients. AB - In the spring and summer of 1988, two separate outbreaks of an illness with a rash resembling erythema infectiosum occurred among members of the nursing staff of the Children's Hospital of Philadelphia. The sources were two adolescent patients with sickle cell disease and aplastic crisis who had unsuspected parvovirus infection. Tests for IgM and IgG antibodies to parvovirus B19 were positive in both patients, and electron microscopical examination showed parvovirus-like particles in the early serum samples. Of 40 health care workers exposed to infected patients, 12 (30 percent) were infected, 2 (5 percent) were possibly infected, 8 (20 percent) had evidence of a past infection with B19, and 18 (45 percent) remained seronegative. Attack rates among the susceptible contacts were 36 percent in the first outbreak and at least 38 percent in the second. Clinical symptoms began a mean of 12.6 days after exposure and included malaise, rash, and joint pain. We conclude that hospital workers are at risk of contracting nosocomial erythema infectiosum from patients with parvovirus associated aplastic crisis. We recommend that all patients with hereditary hemolytic anemias who are admitted with a febrile illness be evaluated for aplasia and promptly placed in respiratory and contact isolation if aplastic crisis is suspected. PMID- 2548097 TI - Hyponatremia and inappropriate secretion of vasopressin (antidiuretic hormone) in patients with hypopituitarism. AB - Severe hyponatremia occurs in some patients with untreated hypopituitarism, but it is not known whether such hyponatremia is caused by the hypersecretion of vasopressin (antidiuretic hormone). This report describes severe, symptomatic hyponatremia in five women 59 to 83 years old (serum sodium, 111 to 118 mmol per liter) who presented with hypopituitarism (which had been previously undiagnosed in four). Plasma vasopressin was inappropriately high (1.3 to 25.8 pmol per liter [1.4 to 28 ng per liter]) in relation to plasma osmolality (236 to 260 mOsm per kilogram of body weight). All five patients had normal renal function and no signs of dehydration or volume depletion. The hyponatremia was resolved within a few days after the institution of hydrocortisone therapy, after infusion of normotonic or hypertonic saline had been found to be less effective. When four of the patients were later restudied while receiving maintenance hydrocortisone treatment, the relation between plasma vasopressin and osmolality was normal. We conclude that ACTH deficiency may cause the syndrome of inappropriate secretion of antidiuretic hormone. The beneficial effect of hydrocortisone is probably exerted through the suppression of vasopressin secretion. PMID- 2548098 TI - Pure red-cell aplasia of 10 years' duration due to persistent parvovirus B19 infection and its cure with immunoglobulin therapy. PMID- 2548099 TI - Human parvovirus B19. PMID- 2548100 TI - Origin of Reed-Sternberg cells in Hodgkin's disease. PMID- 2548101 TI - HTLV-1 myeloneuropathy in the Solomon Islands. PMID- 2548102 TI - [Diffuse cerebral gliomatosis. Clinical, nuclear magnetic resonance tomography and pathologic findings]. PMID- 2548103 TI - [Computerized tomography of the elbow in ulnar sulcus syndrome]. PMID- 2548104 TI - Entropy as a factor in the binding of gamma-aminobutyric acid and nipecotic acid to the gamma-aminobutyric acid transport system. AB - Nipecotic acid is one of the most potent competitive inhibitors and alternative substrates for the high-affinity gamma-aminobutyric acid transport system in neurons, but the structural basis of this potency is unclear. Because gamma aminobutyrate is a highly flexible molecule in solution, it would be expected to lose rotational entropy upon binding to the transport system, a change which does not favor binding. Nipecotic acid, in contrast, is a much less flexible molecule, and one would expect the loss of conformational entropy upon binding to be smaller thus favoring the binding of nipecotic acid over gamma-aminobutyric acid. To investigate this possibility, the thermodynamic parameters, delta G degrees, delta H degrees, and delta S degrees, were determined for the binding of gamma aminobutyrate and nipecotic acid to the high affinity GABA transport system in synaptosomes. In keeping with expectations, the apparent entropy change for nipecotic acid binding (112 +/- 13 J.K-1) was more favorable than the apparent entropy change for gamma-aminobutyric acid binding (61.3 +/- 6.6 J.K-1). The results suggest that restricted conformation per se is an important contributory factor to the affinity of nipecotic acid for the high-affinity transport system for gamma-aminobutyric acid. PMID- 2548105 TI - Purification and characterization of a benzodiazepine-like substance from mammalian brain. AB - An endogenous brain ligand which competes with [3H]-flunitrazepam for the binding to benzodiazepine receptor has been isolated and purified to homogeneity. The purification procedures involve the extraction of the endogenous ligand by homogenizing the brain tissue in water containing various protease inhibitors followed by filtration through a PM 10 membrane (exclusion limit: 10,000-dalton), column chromatographies on Sephadex G-50, Bio-Rad P2 and a series of C18 reverse phase HPLC columns. The purified endogenous ligand was eluted as a single and symmetrical peak monitored at either 220 or 280 nm. Furthermore, the ligand activity coincided with the absorption peak. The purified endogenous ligand is thermostable, insensitive to various peptidases and proteolytic enzymes, resistant to DNAse, RNAse, and carbohydrate enzyme e.g. neuraminidase (EC 3.2.1.18) and acid treatment. It has a major absorption peak at 220 nm and a minor one at 313 nm. The endogenous ligand appears to be quite specific since it only inhibits the binding of ligand to the central type benzodiazepine receptor but not to other receptors, e.g. peripheral type benzodiazepine receptor, alpha 1 adrenoceptor, alpha 2-adrenoceptor, beta-adrenoceptor and muscarinic cholinergic receptor. Furthermore, the inhibition of the receptor binding by the endogenous ligand is enhanced by GABA suggesting that the endogenous ligand is a benzodiazepine receptor agonist. The structure of the endogenous ligand is unknown. PMID- 2548106 TI - Cell-specific fatty acylation of proteins in cultured cells of neuronal and glial origin. AB - Distinct sets of cellular proteins were labeled with [3H]myristic and [3H]palmitic acids in primary (rat neurons and astroglia) and continuous (murine N1E-115 neuroblastoma and rat C6 glioma) cell cultures derived from the nervous system. Both soluble and membrane proteins were modified by myristate in a hydroxylamine-stable (amide) linkage, while palmitoylated proteins were ester linked and almost exclusively membrane bound. Chain elongation of both labeled fatty acids prior to acylation was observed, but no protein amide-linked [3H]myristate originating from [3H]palmitate was detected. Fatty acylation profiles differed considerably among most of the cell lines, except for rat astroglial and glioma cells in which myristoylated proteins appeared to be almost identical based on SDS gel electrophoresis. An unidentified 47 kDa myristoylated protein was labeled to a significantly greater extent in astroglial than in glioma cells; the expression of this protein could be related to transformation or development in cells of glial origin. PMID- 2548107 TI - Evidence for the presence of diacylglycerol kinase in rat brain myelin. AB - The previous demonstration that incubation of brain slices with [32P]phosphate brings about rapid labeling of phosphatidic acid in myelin suggests that the enzyme involved should be present in this specialized membrane. DAG kinase (ATP:1,2-diacyglycerol 3-phosphotransferase, E.C. 2.7.1.107) is present in rat brain homogenate at a specific activity of 2.5 nmol phosphatidic acid formed/min/mg protein, while highly purified myelin had a much lower specific activity (0.29 nmol/min/mg protein). Nevertheless, the enzyme appears to be intrinsic to this membrane since it can not be removed by washing with a variety of detergents or chelating agents, and it could not be accounted for as contamination by another subcellular fraction. Production of endogenous, membrane associated, diacylglycerol (DAG) by PLC (phospholipase C) treatment brought about translocation from soluble to particulate fractions, including myelin. Another level of control of activity involves inactivation by phosphorylation; a 10 min incubation of brain homogenate with ATP resulted in a large decrease in DAG kinase activity in soluble, particulate and myelin fractions. PMID- 2548108 TI - Reconstitution of neurocupreins by Cu-thioneins. AB - Cu-thioneins isolated from liver and brains are able to transfer their copper atoms to apoforms of neurocuprein 1 and 2. At the same time, the apoform of Cu thionein is unable to accept copper from holoforms of both neurocupreins. PMID- 2548109 TI - Efflux of rubidium in rat cortical synaptosomes is blocked by sigma and dextromethorphan binding site ligands. AB - Large concentrations of potassium were used to stimulate the release of rubidium 86 from preloaded cortical synaptosomes, so that the pharmacological sensitivity of this efflux could be examined. Potassium channel blockers, 4-aminopyridine and tetraethylammonium, inhibited the evoked release of rubidium. Sigma ligands, e.g. pentazocine, cyclazocine, rimcazole, 1,3-di(2-tolyl)guanidine (DTG) and haloperidol, as well as the antitussives, carbetapentane, caramiphen and dextromethorphan, significantly reduced potassium-stimulated efflux of rubidium. By contrast, 3-hydroxyphenyl-propylpiperidine (3-PPP), 5-methyl-10,11-dihydro-5H dibenzo(a,d)-cyclohepten-5,10-imine (MK-801), phencyclidine (PCP), ketamine and D 2-amino-5-phosphonovalerate (D-AP5) were all inactive. This suggests that inhibition of potassium-stimulated efflux of rubidium is correlated with activity at the sigma and/or dextromethorphan binding sites rather than at the N-methyl-D aspartate (NMDA)/PCP receptor-channel complex. PMID- 2548110 TI - The noncompetitive N-methyl-D-aspartate receptor antagonist, MK-801 profoundly reduces volatile anesthetic requirements in rabbits. AB - Rabbits anesthetized with volatile anesthetics were given bolus doses of the n methyl-D-aspartate (NMDA) receptor antagonist MK-801. Following observation and recording of the hemodynamic and electroencephalographic effects of MK-801, the animals were tested for requirements of volatile anesthetic to prevent movement to a noxious stimulus. It was demonstrated that MK-801 significantly reduced anesthetic requirements in a dose-dependent manner, while also affecting hemodynamics and the electroencephalogram in a manner consistent with the production of a deeper plane of anesthesia. PMID- 2548111 TI - The effects of GABAB receptor agonists and antagonists on potassium-stimulated [Ca2+]i in rat brain synaptosomes. AB - The effects of GABAB agonists and putative antagonists on intrasynaptosomal calcium ion concentrations ([Ca2+]i) after stimulation with potassium ions were studied with the fluorescent probe Quin 2. gamma-Aminobutyric acid and (-) baclofen, but not (+)-baclofen, produced a dose-dependent inhibition of the potassium-stimulated [Ca2+]i in cortical synaptosomes from the rat. This effect was mimicked by another GABAB agonist SL75102 and weakly by muscimol. It was not inhibited by the alpha-adrenoceptor antagonist phentolamine. This system thus appears to provide a useful test of GABAB receptor function. None of the putative GABAB antagonists, phaclofen, delta-aminovaleric acid or beta-phenyl GABA inhibited responses to (-)-baclofen. Indeed, all three compounds produced similar responses to that seen with (-)-baclofen, suggesting that they act as agonists in this system. These data suggest that those GABAB receptors modulating [Ca2+]i have a distinct pharmacology from post-synaptic GABAB receptors, defined in electrophysiological experiments. PMID- 2548112 TI - Effects of cortisol on GABAA receptor-mediated responses compared in guinea-pig ileum and rat cuneate nucleus. AB - Submaximal contractions of the isolated guinea-pig ileum by the GABAA receptor agonist muscimol were enhanced by about 30% in the presence of 1-100 pM cortisol. At 10 nM cortisol, the enhancement was much less and, at 1 microM, the responses to muscimol were reduced by about 50%. Corticosterone had similar effects but it was 100-1000 times less potent. In contrast, submaximal depolarization responses of the slice preparation of the cuneate nucleus of the rat to muscimol were unaffected by 1 pM-1 nM cortisol and only slightly enhanced (about 15%) by 1 microM cortisol. Also, enhancements of 70-120% by 1 microM alphaxalone were undiminished in the presence of 1 microM cortisol. Thus, there is no evidence for a potent effect of cortisol on the GABAA receptor complex in the cuneate nucleus of the rat. PMID- 2548113 TI - The effect of crotoxin on the longitudinal muscle-myenteric plexus preparation of the guinea pig ileum. AB - The effects of crotoxin, the neurotoxin of the venom of the South American rattlesnake (Crotalus durissus terrificus), was studied by using the myenteric plexus-longitudinal muscle preparation of the guinea pig ileum. Crotoxin (0.02 4.0 microM) caused depression of the twitch response of the electrically stimulated preparation. This transitory depression depended on the concentration of crotoxin; since crotoxin diminished the output of acetylcholine, this depression may be due to the inhibition of the release of acetylcholine from the plexus. Crotoxin also induced an early contraction, followed by relaxation; as the contraction was inhibited by aspirin and indomethacin, it may have resulted from the release of prostaglandin. In addition, a late persistent contracture was observed after the early contraction. The contracture was resistant to blockage by muscarinic, histamine and serotonin antagonists, to hexamethonium, a non depolarizing ganglionic blocking substance and to tetrodotoxin, a sodium channel blocker. The contracture was blocked by an elevated concentration of calcium (10 mM) and by verapamil, a calcium channel blocker. PMID- 2548114 TI - The relationship between anticonvulsant activity and receptor affinity of N methyl-D-aspartate antagonists in epileptic fowl. AB - The N-methyl-D-aspartate (NMDA) receptor antagonists [(3-(+/-)2-carboxypiperazin 4-yl)-propyl-l-phosphonic acid (CPP), +/- 2-amino-7-phosphonoheptanoic acid (2AP7), +/- 2-amino-5-phosphonovaleric acid (2AP5), D-alpha-aminoadipic acid (alpha AA), and +/- alpha, epsilon-diaminopimelic acid (DAP)] were tested for anticonvulsant activity in epileptic chickens. There was a high correlation between anticonvulsant potencies (ED50) and the affinity for the NMDA receptor measured by displacement of L-[3H]glutamate from synaptosomal membranes. The high seizure susceptibility is not due to abnormalities in the NMDA receptor as comparison of KD, Bmax and Ki values in synaptosomal preparations from epileptic and non-epileptic chickens indicated no differences in NMDA receptor binding receptor characteristics. PMID- 2548115 TI - Normal pattern of labeling of cerebral cortical corticotropin-releasing factor (CRF) receptors in Alzheimer's disease: evidence from chemical cross-linking studies. AB - Recent data have demonstrated that in Alzheimer's disease, the concentrations of corticotropin-releasing factor (CRF) were reduced and that there were reciprocal increases in CRF receptors in affected cerebrocortical areas. In order to determine whether the increases in CRF receptors in Alzheimer's disease were due to altered molecular composition of the binding protein, we compared the labeling pattern of 125I-Tyr0-ovine CRF in temporal neocortex of Alzheimer's patients and age-matched controls using chemical cross-linking techniques. A similar pattern of 125I-Tyr0-ovine CRF labeling was seen in Alzheimer's and control brains, with the major CRF binding protein corresponding to an apparent molecular weight of 58,000 Da. These data indicate that the increased CRF receptor population in cerebral cortex in Alzheimer's disease comprises bona fide CRF receptor binding subunits with no apparent change in the molecular structure. PMID- 2548116 TI - Vasopressin (DDAVP) therapy in chronic schizophrenia: effects on negative symptoms and memory. AB - Ten chronic undifferentiated schizophrenics, 6 men and 4 women, aged 28-63, with 6- to 31-year histories of the disease were given DDAVP to observe the effects of this neuropeptide on the prevalent negative symptoms of their illness. Patients were maintained on neuroleptic therapy and first given a 20-day course of placebo followed by 20 days of DDAVP i.m., 4 micrograms Andreasen Scale for assessment of negative symptoms, the Brief Psychiatric Rating Scale, the NOSIE Rating Scale and the Luria-Nebraska Rating Scale were administered to monitor negative symptomatology, behavior and memory before the study began, after placebo and after DDAVP administration. Patients were also given a growth hormone-clonidine test and in addition plasma basal concentrations of 3-methoxy-4 hydroxyphenylglycol (MHPG), homovanillic acid, 3,4-dihydroxyphenylacetic acid (DOPAC) and 5-hydroxyindoleacetic acid (5-HIAA) were measured at the same intervals. DDAVP therapy induced a significant improvement of negative symptomatology and a trend toward improvement of short- to medium-term memory. No changes in homovanillic acid, MHPG, 5-HIAA and DOPAC, nor of growth hormone response to clonidine stimulation were observed. PMID- 2548117 TI - Dose- and time-dependent action of morphine, tramadol and flupirtine as studied by radioelectroencephalography in the freely behaving rat. AB - The necessity of testing psychoactive drugs in awake freely moving animals has led to the development of a telemetry-based system which enables the pharmacologist to follow centrally active molecules in their time- and dose dependent effects on electric brain activity in terms of changes in spectral power density of extracellularly recorded field potentials (tele-EEG). This report describes the effect of three analgesics with respect to bioelectric changes in frontal cortex, thalamus, striatum and reticular formation. Two opiate drugs, morphine and tramadol, behaved very similarly despite a tenfold difference in dosage, whereas flupirtine, a nonopiate analgesic, changed the frequency content of the EEG signals in an entirely different manner. The frequency pattern produced by the opiates closely resembles that of centrally acting serotonin uptake inhibitors and thus is consistent with the view of a serotonergic prevalence of neurochemical interactions within the recorded brain areas. In contrast, the action of flupirtine obviously can be attributed to a clonidine like effect on noradrenergic alpha 2-receptors. The results are discussed with respect to already known influences of these drugs on indoleaminergic and catecholaminergic transmission. PMID- 2548118 TI - Spinal lamina I projection neurons in the rat: collateral innervation of parabrachial area and thalamus. AB - A major ascending nociceptive pathway from spinal lamina I to the mesencephalon has previously been reported in the cat, rat and monkey. In the present paper, we have used single and double retrograde labeling techniques to describe this projection system and its collateralization to the thalamus in the rat. Injections of wheat germ agglutinin-horseradish peroxidase into the pontomesencephalic parabrachial area labeled cell bodies bilaterally in lamina I and deeper laminae of the spinal cord. Bilateral lesions of the dorsolateral funiculi at thoracic levels reduced labeling of lamina I neurons caudal to the lesions. Combined injections of fluorescent retrograde tracers into the lateral thalamus and parabrachial area resulted in double labeling of projection neurons in lamina I, lamina IV VIII and the lateral spinal nucleus of the cervical and lumbar enlargements. Double-labeled neurons were especially abundant in lamina I. Thus, we have demonstrated a major lamina I projection through the dorsolateral funiculi to the parabrachial area with significant collateralization to the thalamus. Moreover, since more than 80% of retrogradely labeled lamina I spinothalamic tract cells had collaterals to the parabrachial area we have indirectly demonstrated the presence of a dorsolateral funicular pathway for lamina I spinothalamic neurons in the rat. More lamina I neurons were retrogradely labeled from midbrain injections as compared to thalamic injections. The significance of these findings rest on previous work in this and other laboratories and concerns the understanding of spinal nociceptive mechanisms. Lamina I projection neurons are primarily nociceptive-specific in their response properties and have been shown to project to both the midbrain and thalamus via the dorsolateral funiculus in a number of species. The role of this projection system in nociceptive transmission may lie in its ability to distribute precise information to multiple brain stem sites which in turn activate autonomic or affective responses or descending pain modulatory mechanisms. PMID- 2548119 TI - Treatment of infantile spasms with high-dose ACTH: efficacy and plasma levels of ACTH and cortisol. AB - Fifteen children with infantile spasms and a hypsarrhythmic EEG defined by EEG videotelemetry monitoring received a regimen of high-dose (150 IU/m2/d) ACTH for their seizures. We carried out an endocrinologic evaluation before and after initiation of the ACTH and conducted a time course study of plasma ACTH and cortisol levels after ACTH dosing. Spasms were controlled and the EEG normalized in 14 of the 15 children. Prior to starting ACTH therapy all the patients had normal prolactin, insulin, cortisol, and ACTH levels in plasma and normal thyroid function. Although the pattern of rise of ACTH levels in plasma after ACTH dosing was similar in all the children, there was great individual variation in the absolute concentrations. However, both the pattern of rise and absolute level of cortisol in plasma after ACTH was highly predictable in all patients. Plasma cortisol rose rapidly within 1 hour of ACTH administration and continued a slower rise for 12 to 24 hours after the ACTH dose. High-dose ACTH therapy seems quite effective in infantile spasms, perhaps because of a sustained high level of plasma cortisol. This sustained plateau of cortisol may be more effective in controlling infantile spasms than the pulse effect expected with oral steroids or lower doses of ACTH. PMID- 2548120 TI - Man-in-the-barrel syndrome caused by cerebral metastases. PMID- 2548121 TI - Electrodiagnosis of brachial plexopathies. PMID- 2548122 TI - [Indications and limitations of subcutaneous mastectomy in the treatment of breast carcinoma]. AB - The reasons that led to subcutaneous mastectomy in some initial stage breast cancer patients are reported. After removal of the mammary gland, this surgical technique provides for prosthetic reconstruction and should therefore be confined to women with initial stage cancer who for psychological or aesthetic reasons refuse more destructive operations. The technique is described and results obtained discussed, recalling that this type of non-destructive operation will be increasingly in demand in the future but needs preclinical diagnosis of the local disease and its systemic diffusion. PMID- 2548123 TI - [Role of computerized tomography in Poland syndrome]. AB - A morphological CT study in Poland syndrome is presented. The osteomuscular anomalies of the chest, which are not demonstrated by usual x-Ray, are well defined by CT scans. The usefulness of CT in postoperative controls of muscles transfers is stressed. PMID- 2548125 TI - Varicella in an immunocompromised patient an electron microscopic study. AB - We report the electron-microscopic appearances of tissues from an immunocompromised patient who died of chickenpox. We observed structures consistent with some of the reported appearances of Varicella-Zoster virus in cultured cells, and tubular structures that have not to our knowledge been previously described. PMID- 2548124 TI - [Chemico-physical property and bile acid binding capacity of several antacids]. AB - Liquid alginate (Gaviscon) binds small amount of bile acids. At pH 7 its viscosity (at low shear rate) is higher than that of other antiacids. High viscosity reduces the diffusion rate of bile salts and glucose and this property can play a role in the treatment of gastro-esophageal and duodeno-gastric refluxes. PMID- 2548126 TI - Physiological evidence for alpha 1-adrenergic facilitatory control of the cold induced TRH release in the rat, obtained by push-pull cannulation of the median eminence. AB - The alpha-adrenergic antagonists phentolamine and prazosin were administered to male rats to explore their effects on cold-induced TRH release, measured by a chronic push-pull cannula stereotaxically implanted in the median eminence (ME). Phentolamine was given either i.p. (24 or 40 mg/kg), or locally (10(-5) M) in the ME, whereas prazosin was only applied locally (10(-5) M). Phentolamine significantly decreased the cold response (5 +/- 1 pg/15 min vs 21 +/- 5 pg/15 min; P less than 0.02), whatever the administration mode. Moreover, the blocking effect of prazosin directly perfused into the ME (11 +/- 3 pg/15 min vs 26 +/- 9 pg/15 min; P less than 0.05), indicates the specific involvement of alpha 1 adrenergic receptors in the cold-induced TRH response, and points to the ME as a possible site of facilitatory adrenergic control. PMID- 2548127 TI - Phosphorylation of mu-opioid receptors--a putative mechanism of selective uncoupling of receptor--Gi interaction, measured with low-Km GTPase and nucleotide-sensitive agonist binding. AB - The mu-opioid receptor agonist stimulation of low-Km GTPase in rat striatal membranes was abolished by islet-activating protein (IAP) treatment, and recovered by Gi reconstitution. When the IAP-treated membranes were phosphorylated with a cAMP-dependent protein kinase, there was no such recovery by Gi. The agonist binding was not affected with respect to Kd, Bmax and sensitivity to guanine nucleotides in the phosphorylated membranes. These findings suggest that phosphorylation of mu-opioid receptors dissociates the agonist change in G-protein activity from the guanine nucleotide-sensitive agonist binding. PMID- 2548128 TI - Coexistence of acetylcholinesterase-, human growth hormone-releasing factor(1-37) , alpha-melanotropin- and melanin-concentrating hormone-like immunoreactivities in neurons of the rat hypothalamus: a light and electron microscope study. AB - Using an antiserum (AS) raised against rat cerebral acetylcholinesterase (AChE), we revealed a neuron population in lateral and dorsal areas of the posterior rat hypothalamus. These neurons were previously described using antibodies to human growth hormone-releasing factor(1-37) (GRF-37), alpha-melanotropin (alpha-MSH) and melanin-concentrating hormone (MCH). Different intracytoplasmic distributions of the immunodeposits were observed depending on the used serum. Ultrastructural investigations demonstrated that AChE-AS labeled rough endoplasmic reticulum and nuclear envelope in control rats. MCH-AS stained Golgi apparatus in control animals and secretory granules in colchicine-injected rats. GRF-37-AS always revealed secretory granules, and alpha-MSH-AS gave the same staining only after colchicine injection. PMID- 2548129 TI - Sodium nitroprusside and guanosine 3',5'-monophosphate (cyclic GMP) inhibit stimulated phosphoinositide hydrolysis in rat cerebral cortical slices. AB - The combination of carbachol and 20 mM K+ greatly stimulates phosphoinositide hydrolysis in rat cerebral cortical slices. This response was inhibited by sodium nitroprusside, an activator of guanylate cyclase which elevated guanosine 3',5' monophosphate (cyclic GMP) levels. 8-Bromocyclic GMP and the phorbol ester phorbol myristate acetate (PMA) also inhibited phosphoinositide hydrolysis, and the inhibitory effect of PMA was additive with that of sodium nitroprusside. As guanylate cyclase is Ca2+-dependent this inhibition of phosphoinositide hydrolysis by cyclic GMP may serve as a feedback modulator of the production of inositol phosphates and the ensuing rise of intracellular calcium. PMID- 2548130 TI - The presence of tau distinguishes Lewy bodies of diffuse Lewy body disease from those of idiopathic Parkinson disease. AB - The antigenic components of Lewy bodies in the cerebral cortex and substantia nigra in 5 cases of diffuse Lewy body disease were examined by immunocytochemistry, using antibodies to neurofilaments (in the phosphorylated or non-phosphorylated forms); to ubiquitin; to the microtubule-associated proteins MAP1, MAP2 and tau; to isolated Alzheimer paired helical filaments, and to tubulin, in the tyrosinated and non-tyrosinated forms. Immunoreactivity with antibodies to cytoskeletal components was identical to that previously described for Lewy bodies of idiopathic Parkinson disease, with the exception that the inclusions of diffuse Lewy body disease (in both cortex and substantia nigra) were stained by an antibody to tau protein. Our findings indicate that although the inclusions found in diffuse Lewy body disease share structural and epitopic features with the inclusions of idiopathic Parkinson disease, they also have distinguishing characteristics (in addition to the differing neuronal populations involved). Also, they suggest that although the inclusions in both conditions appear similar, they probably have different pathogenetic origins. PMID- 2548132 TI - Typhlitis. PMID- 2548131 TI - Melatonin binding sites in the sheep pars tuberalis. AB - The localization of [125I]melatonin binding sites has been studied by autoradiography on frozen unfixed sections of the pituitary stalk, the suprachiasmatic area, the pineal and the pituitary glands in sheep. Dense specific labelling has been found exclusively in the pars tuberalis of the pituitary stalk but not in the part of the median eminence surrounded by the pars tuberalis. The labelling was completely excluded by a 200-fold excess of cold melatonin. No comparable labelling was found in the suprachiasmatic nucleus, the pineal and the pituitary glands. These results constitute the first report of melatonin-specific labelling in the ovine species. PMID- 2548133 TI - Human parvovirus infections--a haematological perspective: any relevance to New Zealand health? PMID- 2548134 TI - Mixed mullerian tumors of the uterus: a clinicopathologic study. AB - Forty-seven cases of mixed mullerian tumors of the uterus were analyzed clinically and pathologically. All patients but one were postmenopausal. Vaginal bleeding was the most frequent presenting symptom, followed by abdominal mass and pelvic pain. Long-term survival was found only in those cases in which the tumor was localized to the uterus (surgical stage I), particularly if it arose from a benign endometrial polyp. No correlation could be established between survival and tumor size, depth of myometrial invasion, or histologic type of sarcoma. Tumors arising after previous irradiation had a poor prognosis. Treatment included surgery, radiation, and chemotherapy. The cumulative probability of 5 year survival was 35%. PMID- 2548135 TI - A prospective 1-year study of estrogen and progestin in postmenopausal women: effects on the endometrium. AB - The endometrial response to oral, cyclic conjugated equine estrogens with and without the randomized addition of medroxyprogesterone acetate was evaluated in 95 postmenopausal women with respect to morphology and bleeding patterns. At 1 year, therapy with 0.625 mg conjugated equine estrogens or 1.25 mg conjugated equine estrogens for 25 days of a 30-day cycle and 5 mg medroxyprogesterone acetate added to the last 11 days of the conjugated equine estrogens cycle was associated with hyperplasia in 0 and 10% of the patients, respectively (P = not significant). Hyperplasia developed in 30 and 57% of the patients who received the above conjugated equine estrogens and placebo regimens, respectively. Irregular, breakthrough bleeding occurred in 14% of the conjugated equine estrogens/medroxyprogesterone acetate users and in 54% of the conjugated equine estrogens/placebo users. The results of this study indicate that 1 year of therapy with 0.625 mg conjugated equine estrogens and 5 mg medroxyprogesterone acetate provided the most satisfactory endometrial protection against hyperplasia and was associated with relatively low rates of breakthrough bleeding. PMID- 2548136 TI - A new and sensitive method of screening for human papillomavirus infection. AB - Human papillomaviruses (HPVs) have been implicated in the development of cervical cancer, and HPV screening may one day supplement cytologic analysis of cervical samples. Detection of the small amounts of HPV DNA in cervical scrapes has been very difficult. The polymerase chain reaction is a new technique that can specifically amplify target DNA to facilitate its detection. We have amplified a region of HPV type 16 DNA using this technique and have shown the method to be both specific and sensitive (a tenfold improvement when compared with Southern blotting). Among 21 cervical scrapes, five from women with normal smears and 16 from women with dyskaryotic smears, HPV 16 DNA was present in scrapes from two women (40%) with normal smears and in scrapes from 12 women (75%) with dyskaryotic smears. The polymerase chain reaction technique is rapid, requires no radioactive probes, and should be the method of choice in future studies to determine the prevalence of HPV infection of the cervix. PMID- 2548137 TI - Hydramnios associated with congenital mesoblastic nephroma: case report. AB - A case is reported of hydramnios due to excessive fetal urine production associated with congenital mesoblastic nephroma. Rapid growth of a right renal tumor and excessive fetal urine production were detected by measuring the size of the tumor and hourly fetal urine production during the development of hydramnios between 28-34 weeks of gestation. A female infant weighing 2200 g was born at 34 weeks' gestation. She lost 18% of her birth weight in the first 48 hours because of neonatal polyuria. Right nephrectomy and ureterectomy were performed 2 days after birth. After the operation, the neonatal polyuria improved dramatically. The right renal tumor was identified histologically as a mesoblastic nephroma. The importance is stressed of early and correct antenatal diagnosis of a renal tumor and precise estimation of urine production for postnatal management of hemodynamic change in cases of mesoblastic nephroma. PMID- 2548138 TI - Pelvic trophoblastic implants after laparoscopic removal of a tubal pregnancy. AB - Laparoscopic removal of ectopic gestations is becoming increasingly popular. We present a case in which an early, unruptured ampullary ectopic pregnancy was identified clinically, removed during laparoscopy, and subsequently confirmed by pathology. The patient later presented with pain and with rising titers of beta hCG. Laparotomy demonstrated multiple pelvic implants of trophoblastic tissue. PMID- 2548139 TI - [Enhancement of enamel crystallinity with gel method during orthodontic treatment. The 2nd report: application of micro-area X-ray diffraction method]. AB - This study is to review an effect on the improvement of enamel crystallinity with gel method in the course of orthodontic treatment by applying gelatin gel method (group II) as well as silica-hydro gel method (group I) to bracket-bonded enamel surfaces of extracted teeth and for a relative review, the chopped cross sections of the above teeth were analyzed by the method of micro area X-ray diffraction. The results obtained were as follows: 1. As a result of identifying by the method of microarea X-ray diffraction, the present of hydroxyapatite, Ca5(PO4)3OH and fluorapatite, Ca5(PO4)3F was recognized at any section of every group. 2. In making a relative study of the variation of line profile and peak shift, an outstanding difference was not acknowledged in line profile, however, peak shift in the direction of high angle side was indicated on the experimental side of each group and a tendency for peak shift was more noticeable near by enamel cuticle. At boundary sites and on the control side, the variation was only shown in one part of group II. Also a tendency for peak shift in the direction of high angle side was recognized in group I rather than in group II. From the foregoing results, it was found to be effective to improve tooth substance of the bracket bonded enamel and in that event, the silica-hydro gel method was suggested to be superior to the gelatin gel method. Also with the regard to the bracket-bonded sides, tooth substance was noticeably improved at enamel cuticle but not sufficiently improved at sites underneath the brackets and at boundary sites of the both, therefore, one of the future subjects would be to review such areas. PMID- 2548140 TI - [Lymphokine-induced suppression of erythropoiesis by normal T lymphocytes and retrovirus-associated lymphoproliferative disease]. AB - Activation of genes for interleukin 2 (IL2) and its receptor by the tat gene product of the human retrovirus HTLV-I has been linked to the pathogenesis of adult T cell leukemia. In earlier studies we have demonstrated that IL2-induced inhibition of erythropoiesis is mediated by activated T-Lymphocytes expressing the IL2-receptor. We have now examined the role of activated T cells and interferon-gamma (IF gamma) in inhibition of normal erythropoiesis and in HTLV-I associated lymphoproliferative disease. T cells were activated by triggering of the antigen receptor complex with CD3 monoclonal antibody. Incubation with CD5 antibody recognizing a T cell epitope distinct from the antigen receptor served as a control. Supernatants derived from CD3 activated T cells generated in the presence of IL2 caused a dose-dependent suppression of erythropoiesis. Treatment of supernatant interferon-gamma neutralizing antibody caused a greater than 95% abrogation of inhibition. Next we investigated the mechanisms for acquired pure red cell aplasia in a patient with T gamma-lymphoproliferative disease (T gamma LPD). Patient marrow erythroid progenitors were 17 +/- 9% of control and increased to 88-102% of control following marrow T cell depletion. Myeloid progenitors were normal. Patient suppressor T cells inhibited growth of autologous and allogeneic marrow erythroid but not myeloid progenitors and produced high titers of IF gamma. The inhibitory factor in patient T cell supernatant was acid labile and sensitive to trypsin, consistent with properties of IF gamma. Prior depletion of activated T cells abrogated the suppressive effect of patient T cell-derived supernatant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548142 TI - Fibrous histiocytoma of the conjunctiva. AB - Fibrous histiocytomas are commonly found on the skin, whereas the appear to affect the conjunctiva only very rarely. PMID- 2548141 TI - [Mechanism of resistance to tumor necrosis factor]. AB - To find clues as to the possible mechanisms of resistance against the cytotoxic activity of tumor necrosis factor (TNF), we developed TNF-resistant and TNF sensitive subclones of the human cervic carcinoma cell line ME 180 and the mammary carcinoma cell line BT-20. Resistant subclones were selected by gradually increasing the TNF concentration in the culture medium over 4 months or 20 passages. Resistance to TNF was conserved in the absence of TNF after nearly 3 years and less than 100 passages. There were no differences in the number and affinity of TNF receptors between the sensitive and resistant clones. Similarly, SDS-PAGE of cell lysates showed identical bands. Sensitivity to actinomycin D, mitomycin C, and interferon-gamma was the same. No TNF message could be demonstrated in either subclone. As tumors grown in nude mice derived from the sensitive and resistant clones responded equally well to an intratumoral injection of TNF, we conclude that different mechanisms are responsible for sensitivity and resistance to the cytotoxic effect of TNF in vitro and in vivo. PMID- 2548143 TI - N-myc activation by proviral insertion in MCF 247-induced murine T-cell lymphomas. AB - N-myc proto-oncogene rearrangement was found in three out of six AKR murine T cell lymphomas induced by the highly oncogenic MCF 247 MuLV. Molecular analyses showed that structural modification of the proto-oncogene in all three lymphomas was in the consequence of MCF 247 proviral integration within the gene III exon. All integrated proviruses have the same transcriptional orientation as the N-myc gene. As a consequence of proviral insertion, the N-myc gene becomes transcriptionally active, producing an abnormal mRNA. These findings suggest a possible causative role of such an integrative event in murine T-cell lymphomagenesis. PMID- 2548144 TI - Transformation of murine fibroblasts by a retrovirus encoding the murine c-fms proto-oncogene. AB - The c-fms proto-oncogene is the growth factor receptor for the macrophage colony stimulating factor, M-CSF. In this paper we have investigated the inappropriate expression of c-fms within a species as a relevant model for analysing transformation, growth, and differentiation promoting activities of c-fms. A retroviral vector was constructed for the expression of the murine c-fms proto oncogene in murine cells. Initial characterization of this vector was performed in fibroblasts. Infection of Balb/c 3T3 cells with the murine c-fms retrovirus resulted in high level expression of the gp140c-fms precursor and the gp165c-fms mature receptor protein on the cell surface and both proteins exhibited kinase activity. Up to 3.3 x 10(4) M-CSF receptors per cell were detectable. The c-fms retrovirus induced foci of morphologically transformed Balb/c cells that exhibited anchorage independent growth in soft agar and produced progressively growing tumors in syngeneic animals. The Balb/c cells synthesize and secrete M CSF and therefore these results suggest an autocrine mechanism of transformation caused solely by the expression of a normal growth factor receptor in an inappropriate, endogenous M-CSF-producing cell. PMID- 2548145 TI - Regulation of c-fgr proto-oncogene expression in Burkitt's lymphoma cells: effect of interferon treatment and relationship to EBV status and c-myc mRNA levels. AB - The proto-oncogene c-fgr is expressed in transformed human B lymphoid cell lines and has been reported to be induced in cells infected with the Epstein-Barr virus (EBV). We have compared the levels of c-fgr mRNA in several B cell lines and have examined the effects of interferon-induced changes in growth rate of Daudi cells on the concentration of this mRNA. High levels were found in exponentially growing EBV-positive Raji and Daudi cells but the amounts in B95-8 and P3HR-1 cells (also EBV-positive) were lower than in the EBV-negative cell line Ramos. Growth inhibition of Daudi cells by interferon-alpha is preceded by a reduction in c-fgr expression, with a 56% decrease observed within 6 h. The differences in the amounts of c-fgr mRNA in the various cell lines and in control versus interferon-treated cells are similar to the differences in the c-myc mRNA contents of these cells. These results indicate that c-fgr expression bears little relationship to the EBV status of B lymphoid cell lines but may play a role, in conjunction with c-myc expression, in growth regulation by interferons. Other conditions which influence Daudi cell proliferation, such as treatment with a phorbol ester or growth to high cell density, also inhibit c-fgr expression but to a lesser extent than interferon treatment. PMID- 2548146 TI - In situ and in vitro evidence for intragenic binding of nuclear factors at the murine c-myc locus. AB - A block to transcriptional elongation within the c-myc proto-oncogene has been previously observed in a large number of different mouse and human cell types and its release is a potentially important element in the pathogenesis of some malignancies. We show here that the chromatin around the mouse c-myc exon 1 intron 1 boundary is differentially accessible to restriction enzymes in purified nuclei. Using a combination of in situ exonuclease III protection assay with in vitro footprints and gel band shifts, we have shown the existence of a stable nucleoprotein complex in this same region in mouse erythroleukemia cell nuclei. This situation is not peculiar to these cells and we have shown that the accessibility of the two BglII sites present at the beginning of intron 1 seems to depend not only upon the transcriptional state, but also upon the structural integrity of the gene. PMID- 2548147 TI - [Klippel-Trenaunay syndrome, a case report]. AB - Description of the Klippel-Trenaunay-Syndrome; it could have been sporadic found in a 3; 6-year-old girl. This congenital anomaly is characterized by haemangiomatosis, varicosis and unilateral hypertrophia of limbs. As a rule the therapy is a symptomatic one, surgical operations concerning the skeleton are infrequent. PMID- 2548148 TI - Role of hypochlorous acid in Trypanosoma musculi killing by phagocytes. AB - Trypanosoma musculi are readily killed when phagocytosed by mononuclear phagocytes but the nature of the mediators of this cytotoxicity is unclear. Among the most potent mediators are oxygen-derived species. The generation of chemiluminescence (CL) by peritoneal macrophages from 12 day T. musculi-infected mice, which phagocytose and kill parasites when opsonizing antibodies are present, was recorded in the presence of antibody-coated trypanosomes. Taurine, a specific quencher of hypochlorous acid (HOCl) inhibited CL production by peritoneal macrophages, showing that HOCl is produced during phagocytosis of T. musculi. In vitro, HOCl alone exerted a powerful trypanocidal activity which was inhibited in the presence of specific quenchers. The role of HOCl generated by phagocytes in trypanosome killing was studied using granulocytes which produce more oxygen-derived species than macrophages when stimulated. Phorbol myristate acetate-triggered granulocytes can destroy T. musculi and trypanosome killing is inhibited in the presence of taurine. These data demonstrate that HOCl produced by phagocytes can effectively destroy T. musculi. PMID- 2548149 TI - Detection of human papillomavirus DNA in cervical lesions by in-situ DNA hybridization. AB - Authors examined paraffin sections of 50 cervical specimens from 34 cases for the presence of human papillomavirus (HPV) type 6b, 11, 16, 18, 31 and 33 by in-situ hybridization using 35S-labelled HPV DNA probes. Specimens were classified according to the degree of dysplasia after histological examination. Viral nucleic acids were detected in 30 of 50 tissues (60%) in which 15 specimens had single, 10 double, 4 triple and 1 quadruple viral infections. In some cases, different viral nucleic acids were detected at separate sites in the same patient. Overall, no great variation in the frequency of each HPV was detected, but a pattern became apparent when the frequencies were compared with the grade of dysplasia. CIN II/III lesions contained one or more of the HPV types 16, 18, 31, 33 which are frequently associated with cervical carcinoma. In-situ hybridization offers sensitive means of investigating viral infection, gene expression and neoplastic transformation. PMID- 2548150 TI - [Level of pituitary, adrenal and thyroid hormones in parturients with allergy and in their progeny]. PMID- 2548151 TI - [Diagnostic significance of calcitonin analysis in children with rickets and rickets-like diseases administered calcium test]. AB - The secretory activity of C-cells of the thyroid was investigated in 25 children with rickets and rickets-like diseases. There were 13 children with grades II and III rickets, 6 with vitamin D-dependent rickets, and 6 with Debre-de Toni-Fanconi disease. Five normal children constituted a control group. The basal concentration of calcitonin did not differ from the control parameter in all patients' groups. An increased release of the hormone to the blood was noticed in response to calcium stimulation. The rise of blood calcitonin was determined by the disease pattern, which made it possible to use the test with a one-minute intravenous calcium injection for differential diagnosis purposes. The maximal changes were discovered in children suffering from vitamin D-dependent rickets. PMID- 2548152 TI - [Classification of rickets]. PMID- 2548153 TI - Ty4, a novel low-copy number element in Saccharomyces cerevisiae: one copy is located in a cluster of Ty elements and tRNA genes. AB - We have identified a composite element, Ty4, in S. cerevisiae that is ca 6.3 kb in length and contains two tau sequences as long terminal repeats. According to hybridization analyses, Ty4 occurs in low but varying copy number (one to four copies) in different yeast strains. By several criteria, Ty4 is a novel type of retroelement which is similar but not related to the other Ty elements in yeast. Two cosmid clones from strain C836 (c90 and c476) carrying individual copies of Ty4 were isolated. By restriction analysis and nucleotide sequence we show that c476 derives from the 'transposition right arm hot spot' of chromosome III [1]. The analysis of c476 revealed that an initiator tRNA(Met) gene is present at this locus and that an unusual concentration of different Ty elements has occurred: in addition to the Ty4, a Ty1 and a Ty2 element were detected in this region, confirming its highly polymorphic character. PMID- 2548154 TI - Protein-induced unwinding of DNA: measurement by gel electrophoresis of complexes with DNA minicircles. Application to restriction endonuclease EcoRI, catabolite gene activator protein and lac repressor. AB - An electrophoretic procedure for the measurement of the helix unwinding induced by a sequence-specific protein is described. The method, which was applied here to EcoR I, CAP and lac repressor, involved the migration of the complexes with positively and negatively supercoiled DNA minicircles carrying a single protein binding site. Mobility shifts of complexes relative to naked DNAs appeared to be a result of i) the unwinding; of ii) an increase in the molecular frictional coefficient, which led to a retardation; of iii) bending, in the particular case of CAP, which induced an acceleration; and of iv) looping, in the case of lac repressor, which also resulted in an acceleration. Under conditions where the migration of the naked topoisomers was V-like (topoisomer mobility showed the same linear increase with both negative and positive supercoilings; Zivanovic et al. (1986) J. Mol. Biol., 192, 645-660), the protein unwinding contribution to mobility was assumed to be identical to that experimentally observed in the case of a thermal unwinding: all negatively supercoiled topoisomers were retarded and all positively supercoiled topoisomers were accelerated to the same extent. In contrast, the mobility contribution of the frictional term, as well as those of bending and looping, appeared to vary strongly with the magnitude of the supercoiling, but only weakly with its polarity. As a consequence, these latter contributions may approximately cancel when one is measuring the difference between the shifts observed for two comigrating, negatively and positively supercoiled, topoisomers, allowing the unwinding to be calculated. While estimates obtained for EcoR I, 23 +/- 3 degrees, and CAP, about 29 degrees, were in good agreement with previous measurements using topoisomerase I, the value found for lac repressor, 13 to 16 degrees, was significantly smaller. PMID- 2548155 TI - RNAs containing mitochondrial ND6 and COI sequences present an abnormal structure in chemically induced rat hepatomas. AB - We have constructed a cDNA library prepared from an hepatoma cell line (HTC cells) and isolated a clone, pHT 13, which corresponds to mRNAs present at a much higher level in rat hepatomas than in normal hepatocytes. The sequence of the pHT 13 insert has been previously published (Nucleic Acids Res. 1988, 16,10935). This clone contains mitochondrial DNA sequences with an abnormal organization, since it includes part of the NADH dehydrogenase subunit 6 (ND6) and of the cytochrome oxidase subunit I (COI) genes separated by 230 bases instead of 9 kb in mitochondrial genome from normal hepatocytes. In this work we show (1) that RNAs homologous to this clone are present in hepatoma cells but not in normal hepatocytes, (2) that a 3 kb fragment of tumor mitochondrial DNA contains both the ND6 and the COI sequences. The abnormal structure of the DNA is confirmed by Southern blot analysis which shows that distinct types of mitochondrial DNAs are present in hepatoma cells. PMID- 2548158 TI - Primary structure of the SSAV tether--RNase H endonuclease (pol) region deleted in SSV. PMID- 2548156 TI - Differential activation of the 21-base-pair enhancer element of human T-cell leukemia virus type I by its own trans-activator and cyclic AMP. AB - A transcriptional trans-acting factor p40tax of human T-cell leukemia virus type I (HTLV-I) functions as an inducer for expression of HTLV-I provirus via activation of the enhancer in the long terminal repeat of HTLV-I. In addition to p40tax and a tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA), an activator of protein kinase C, we report here that forskolin, an activator of adenyl cyclase, also induces function of the HTLV-I enhancer. Experiments with mutants of the HTLV-I enhancer revealed that TPA-induced activation was not mediated by solely a 21-base-pair (bp) sequence that is repeated three times in the enhancer, whereas the 21-bp enhancer element can act as a sufficient cis acting sequence for activation by both p40tax and forskolin. In addition, we found that nuclear factor(s) like the cyclic AMP-responsive element (CRE) binding factor could bind to the HTLV-I 21-bp enhancer element. However, a difference was found in sequences required for activation by p40tax and forskolin. A CRE related sequence present in the 21-bp enhancer element was enough for forskolin-induced activation. On the other hand, p40tax required a much longer sequence that is overlapping but not identical to the CRE related sequence, suggesting that the forskolin-induced cyclic AMP pathway may be partly involved in, but not sufficient for p40tax-mediating trans-activation of the HTLV-I enhancer. PMID- 2548157 TI - DNase I sensitivity of immunoglobulin light chain genes in Abelson murine leukemia virus transformed pre-B cell lines. AB - We have used Abelson murine leukemia virus (A-MuLV) transformed pre-B cell lines to test the hypothesis that the rearrangement potential of a developing B lymphocyte is dependent on an "opening" of the chromatin structure surrounding immunoglobulin (Ig) genes, thus allowing accessibility to an Ig gene recombinase. The chromatin structures surrounding heavy (H), kappa (kappa), and lambda (lambda) chain constant-region genes were assessed by DNase I sensitivity in A MuLV transformed cell lines capable of H, kappa or lambda gene rearrangement. Our results indicate that DNase I-sensitive chromatin structures of these Ig constant region genes correlate closely with the ability of the genes to undergo recombination. We also find that the chromatin structure of an Ig constant-region locus becomes DNase I sensitive before any DNA rearrangement events occur. PMID- 2548159 TI - A possible relationship of reovirus putative RNA polymerase to polymerases of positive-strand RNA viruses. PMID- 2548161 TI - Effect of lambda packaging extract mcr restriction activity on DNA cloning. PMID- 2548162 TI - DNA extraction with organic solvents in gel barrier tubes. PMID- 2548160 TI - Rapid insertional mutagenesis of DNA by polymerase chain reaction (PCR). PMID- 2548163 TI - High resolution hydroxyl radical footprinting of the binding of mithramycin and related antibiotics to DNA. AB - The preferred binding sites for mithramycin on three different DNA fragments have been determined by hydroxyl radical footprinting. Sequences which appear as one long protected region using DNAase I as a footprinting probe are resolved into several discrete binding domains. Each drug molecule protects three bases from radical attack, though adjacent regions show attenuated cleavage. Mithramycin and the other related compounds induce similar footprinting patterns and appear to recognise GC rich regions with a preference for those containing the dinucleotide step GpG. The ability of each such site to bind the drug depends on the sequence environment in which it is located. The data are consistent with mithramycin binding to the DNA minor groove. PMID- 2548164 TI - Up-promoter mutations in the positively-regulated mer promoter of Tn501. AB - Transcription from the mer promoter of transposon Tn501 is repressed by MerR (the product of the merR gene) in the absence of Hg2+, and activated by MerR in the presence of Hg2+. In the absence of MerR, the mer promoter has weak constitutive activity. The DNA sequence of the mer promoter shows candidate -35 and -10 sequences at the unusually high spacing of 19 base-pairs. We have selected for spontaneous mutations in the mer promoter that confer an up-promoter phenotype. Four different mutants have been isolated. Three of these are single base-pair deletions between the -10 and -35 sequences. A fourth removes the -10 sequence entirely, and places a second potential -10 sequence 17 base-pairs from the -35 sequence. None of these mutant promoters are induced by MerR in the presence of Hg2+. Two of them are repressed by MerR irrespective of the presence or absence of Hg2+. Models for the mode of action of the MerR protein are discussed in the light of these results. Our data support a mechanism in which the MerR protein in the presence of Hg2+ acts to change the conformation of DNA in the mer promoter. PMID- 2548165 TI - Gene transfer of truncated NGF receptor clones leads to cell surface expression in mouse fibroblasts. AB - Transfection of recombinant bacteriophage clones encoding human NGF receptor sequences resulted in cell surface expression in mouse fibroblasts. Unexpectedly, receptors were expressed even after transfection with phage clones which lack 5' gene sequences. Stable transformants were purified and analyzed in detail. S1 nuclease protection and primer extension analysis revealed that an initiation site lies within an intron sequence in the middle of the receptor gene. A truncated mRNA transcript was detected that allowed for the expression of NGF receptors capable of binding to NGF. Since the original phage clones lacked the first two exons, these results suggest that the normal N-terminal sequences may not be necessary for cell surface expression and binding to NGF. PMID- 2548167 TI - Conservation of the secondary structure elements of the 5'-untranslated region of cardio- and aphthovirus RNAs. AB - An analysis of published nucleotide sequences of the 5'-untranslated region (5' UTR) of 7 cardioviruses and 3 aphthoviruses has allowed us to derive a consensus secondary structure model that differs from that previously proposed for the 5' UTR of entero- and rhinoviruses, though all these viruses belong to the same family, Picornaviridae. The theoretical model derived here was experimentally supported by investigating the accessibility of encephalomyocarditis virus RNA to modifications with dimethyl sulfate and its susceptibility to S1 and cobra venom nucleases. The possible involvement of the 5"-UTR secondary structure domains in the translational control is briefly discussed. PMID- 2548168 TI - Nucleotide sequence of the cDNA encoding subunit AED (VIB) of beef heart cytochrome c oxidase. PMID- 2548166 TI - Transcription of the chicken myb proto-oncogene starts within a CpG island. AB - The nucleotide sequence of an 8.2-kb DNA fragment from the 5' proximal part of the chicken myb proto-oncogene spanning 1761 nucleotides upstream and 6436 nucleotides downstream from a presumed c-myb initiation codon was determined. A 3.3-kb G + C-rich region found in this sequence had also other features characterizing CpG islands, i.e. no CpG underrepresentation and lack of CpG methylation. In haematopoietic tissues c-myb mRNA synthesis starts in two major regions of the CpG island, namely 98 to 108 and 143 to 145 nucleotides upstream from the c-myb initiation codon. These two regions are in or close to the 124-bp evolutionarily conserved element located in the middle part of the CpG island. No alternative splicing of the 5' end of c-myb mRNA suggested earlier (1,2) was observed. The c-myb promoter contains neither TATA nor CAAT box-like structures at the usual positions. Instead, numerous potential Sp1 factor binding sites were found both upstream and downstream from the transcription initiation sites. Moreover, consensus v-myb protein DNA-binding sites were revealed in the promoter region and in sequences downstream from it. PMID- 2548169 TI - Hamster line and ALU-equivalent sequences are present in the small polydispersed circular DNA population of CHO cells. PMID- 2548170 TI - Nucleotide sequence of cDNA for murine myeloperoxidase. PMID- 2548171 TI - Production of single-stranded DNA templates by exonuclease digestion following the polymerase chain reaction. PMID- 2548172 TI - Radioreceptor assay of metoprolol in human plasma: comparison with an enantiospecific high-performance liquid chromatographic (HPLC) procedure. AB - Plasma concentrations of metoprolol after acute and repetitive administration of R/S-metoprolol to healthy volunteers were measured by a beta-adrenoceptor subtype specific radioreceptor assay (RRA) and by an enantiospecific high-performance liquid chromatographic (HPLC) method. In the RRA, R/S-metoprolol showed a 20-fold beta 1-subtype selectivity: the S-(-)-enantiomer was 35-fold more potent than the R-(+)-enantiomer. A comparison between S-(-)-metoprolol concentrations detected in the plasma samples by HPLC and those detected by RRA yielded a 1/1 relationship, indicating that active metabolites are not present to a significant extent. These results were independent of the widely scattering metabolic clearance of metoprolol (with the potential of differences in the rate and extent of formation of active metabolites) in the volunteers. In general, HPLC methods can be validated by comparison with RRA in order to clarify whether active metabolites are present and--on the basis of the Ki value from RRA--whether the detection limit of the physicochemical procedure is sufficient to cover the therapeutically relevant range. PMID- 2548173 TI - Synthesis and receptor binding studies of (+/-)1-iodo-MK-801. AB - The glutamate analogue N-methyl-D-aspartate (NMDA) binds to a subset of glutamate receptors that are coupled to a voltage-sensitive cation channel. This NMDA linked channel is the likely binding locus of the potent anticonvulsant MK-801. To develop single-photon emission computed tomography (SPECT) probes of this brain channel, we synthesized (+/)1-iodo-MK-801 and (+/-)1-[125I]iodo-MK-801. The effect of (+/-)1-iodo-MK-801 on ligand binding to the NMDA-linked glutamate receptor site was assessed using a rat brain homogenate assay. (+/-)1-Iodo-MK-801 displaced the dissociative anesthetic ligand [3H]N-[1-(2 thienyl)cyclohexyl]piperidine ([3H]TCP) binding with an IC50 of 1 microM, which is a 10-fold lower binding affinity than that of (+/-)MK-801. In in vivo autoradiographic studies, (+/-)MK-801 failed to block selective uptake of (+/-)1 iodo-MK-801 in rat brain. These results suggest that (+/-)1-iodo-MK-801 may not be a suitable ligand for mapping NMDA-linked glutamate receptor channels. PMID- 2548174 TI - Effect of feeding some plant foods as source of dietary fibre on biological utilisation of diet in rats. AB - Isoproteinous diets containing 1% cholesterol and 10% fibre derived from plant foods namely, peepalbanti (Ficus religiosa), barbanti (Ficus bengalensis), gullar (Ficus glomerata), teent (Capparis decidua) and khejri beans (Prsopsis cineria) or pure cellulose, were fed to rats for 40 days. Effect of incorporation of these whole plant parts on feed intake, weight gain, feed efficiency ratio (FER), dry matter digestibility (DMD) and true protein digestibility (TPD) in weanling rats was studied. Their inclusion did not affect the weight gain significantly whereas all other parameters were influenced to a varying extent. Foods rich in lignin had relatively lower FER, DMD and TPD whereas cellulose and pectin rich foods had higher DMD and TPD. PMID- 2548175 TI - Nutrient composition and protein quality of minor millets. AB - Nutrient composition of five minor millets produced and consumed in dry land regions and tribal areas of Andhra Pradesh, India, was evaluated. The millets analysed were Italian millet (Setaria Italica), French millet (Panicum miliaceum), Barnyard millet (Echinachloa colona), Kodo millet (Paspalum scrobiculatum) and little millet (Panicum miliare). The nutrients analysed were proximate principles, amino acids, fibre components, calcium, Phosphorus and Iron. Protein and energy were evaluated by true digestability (TD), biological value (BV), net protein utilization (NPU) and digestible energy (DE) in rats. The protein content was higher and lysine content lower, when compared to major millets. Dehusked millets had lower total dietary fibre (TDF) and tannin. The TD of protein ranged between 95.0 to 99.3, whereas the BV was between 48.3 and 56.5. All the millets were poor sources of calcium, phosphorus and iron, with the exception of french millet. PMID- 2548176 TI - [Treatment of diabetic ketoacidosis with small doses of insulin]. AB - The authors discussed the results of the treatment of 50 patients who had been admitted to hospital with diabetic ketoacidosis. The purpose of the investigation was to prove the effectiveness and reliability of therapy of diabetic ketoacidosis using small doses of insulin in combination with the administration of a sufficient amount of liquid, potassium and, when indicated, sodium bicarbonate. The authors presented a detailed scheme of therapy of diabetic ketoacidosis and assessed the effectiveness of its use based on analysis of clinical and laboratory findings. The results obtained permitted its recommendation for a wide clinical use. PMID- 2548177 TI - Glucose-stimulated hormone release in rats bearing streptozotocin/nicotinamide induced islet adenomas: evidence for slow and fast responders. AB - Rats injected with streptozotocin and nicotinamide developed grossly visible islet cell tumors of the pancreas. During i.v. glucose tolerance tests, two populations of tumor-bearing rats were identified: fast responders exhibited significantly lower plasma glucose and markedly elevated plasma immunoreactive insulin (IRI) levels relative to those of the controls. In slow responders, the plasma glucose level was significantly elevated up to 2 h after glucose injection, and the plasma IRI level was lower than that of the controls. During in vitro perfusions with glucose at 300 mg/dl (16.7 mM), tumor-bearing pancreata of fast responders released elevated levels of IRI and immunoreactive somatostatin (IRS); after tumor removal, glucose-stimulated release of these hormones returned to control levels. However, during similar perfusions of pancreata from slow responders, the IRI and IRS release did not decrease after tumor removal, suggesting that the nontumorous pancreatic islets rather than the gross tumors of the slow-responder group were the source of the glucose stimulated hormone release. These studies demonstrate that gross tumors in the two responder subgroups differ in their glucose-stimulated hormone release. PMID- 2548178 TI - A malignant tumor of the pancreas producing glucagonoma syndrome: immunocytochemistry and ultrastructure of liver metastases and comparison with the primary tumor. AB - In this study, liver metastases from a patient with a pancreatic glucagonoma producing the syndrome have been investigated histologically, ultrastructurally, and immunocytochemically. A comparison has also been made between the metastases and the primary pancreatic tumor investigated in a parallel study. In the metastatic tissue, glucagon-, pancreatic polypeptide (PP)-, and somatostatin containing cells were found together with a majority of cells without any immunoreactivity. Glucagon-positive cells were much more numerous than PP- and somatostatin-immunoreactive cells. As in the primary tumor, double immunogold staining of ultrathin sections demonstrated the co-existence of glucagon and PP immunoreactivities in most of the granulated cells, but PP immunolabeling was often faint, so that it probably could not be revealed by the PAP method in light microscopical sections. Such a finding, together with the histological and ultrastructural features, is consistent with an ontogenic and phylogenetic primitiveness of the metastatic cell population. PMID- 2548179 TI - A malignant tumor of the pancreas producing glucagonoma syndrome: coexistence of glucagon and pancreatic polypeptide (PP) in the tumor cells. AB - A malignant tumor of the pancreas producing the glucagonoma syndrome and associated with high plasma levels of glucagon and pancreatic polypeptide was studied histologically, ultrastructurally, and immunocytochemically. The histologic and ultrastructural features were closely similar to those of previously reported malignant glucagonomas. However, immunolabeling with specific antisera revealed that, in addition to cells having only glucagon- or only pancreatic polypeptide-immunoreactivity, other cells were also present, showing a co-existence of both peptides. These findings indicate that the tumor contains a cell population with a phenotype similar to that of intestinal L-cells rather than to pancreatic A-cells. PMID- 2548180 TI - Pigment resembling atmospheric dust in Peyer's patches. AB - Terminal ileal pigmentation was observed during colonoscopy, in surgically resected specimens, and autopsy cases. Microscopically, black pigment was seen within macrophages in the lamina propria and submucosa, closely related to the Peyer's patches. Three ilia from autopsies with no macroscopic pigmentation showed deposits following digestion and X-ray microanalysis. X-ray microanalysis of tissue sections and digestates revealed a heterogenous population of particles. Approximately one third of the particles contained calcium and phosphorus and were considered endogenous. The rest of the particles were predominantly aluminum and magnesium-rich silicates, which were considered exogenous. Analysis of particulate extracted from lungs and ilea of four autopsy cases demonstrated remarkable similarities in composition. These findings suggest that the ileal deposits are derived from atmospheric dust. This pigment is believed to migrate into the Peyer's patches through the M cells of the follicle associated epithelium, although other mechanisms for pigment deposition cannot be ruled out. PMID- 2548181 TI - Primary linitis plastica carcinoma of the colon and rectum. AB - Linitis plastica carcinoma of the colon and rectum is a relatively rare primary colorectal malignancy. The initial diagnosis of this disease is often difficult, and its eventual prognosis is poor. In this report, we describe a case of primary linitis plastica of the rectum in a 57-yr-old American Indian. This case is unique since it represents the first documentation of a primary rectal lesion with metastases to the stomach. The current literature is reviewed with examination of the clinicopathologic features of this neoplasm. PMID- 2548182 TI - Usefulness of combined light and electron microscopy: evaluation of sputum samples for asbestos to determine past occupational exposure. AB - Ferruginous bodies (FB) in sputa are recognized as an indicator of past exposure to asbestos. However, a great variability exists in FB production, even in individuals with a history of occupational exposure. A further complication in interpreting the presence of FBs in sputa is that all individuals in modern society are exposed to asbestos and, in lung tissue studies, have been shown to harbor appreciable numbers of asbestos fibers. Thus, some of these individuals should occasionally produce FBs in their sputa. The present study was undertaken to determine if uncoated asbestos fiber content could be used to better discriminate occupationally exposed individuals from the general population. Randomly selected sputum samples from 12 former workers in an amosite asbestos plant and 12 controls were studied. The samples were prepared for the study by digesting the sputa in sodium hypochlorite. The digests were filtered through 0.2 microns polycarbonate filters for collection of particulates. The filters were screened for FBs by light microscopy at 200 X, and the presence or absence of uncoated asbestos fibers was determined at 5000 X in an AMRAY 1000A scanning electron microscope. The use of electron microscopy revealed the presence of commercial amphiboles in the sputa of the occupationally exposed individuals and enabled a differentiation of these samples from those of the general population. PMID- 2548183 TI - Mucosal origin of sinonasal tract adenomatous neoplasms. AB - One hundred seven sinonasal tract adenomatous neoplasms on file at the Armed Forces Institute of Pathology were reviewed in order to determine what percentage of and what type of nonsquamous epithelial neoplasms arise from surface mucosa. Seventy-seven percent of tumors were localized to the nose or maxillary sinus. Fifty-nine of the 107 tumors (55%) including 32 (50%) of the 64 adenocarcinomas and 19 (59%) of the 31 adenoid cystic carcinomas arose from the surface mucosa. Five of the six mucoepidermoid carcinomas also appeared to arise from the surface in a diffuse fashion, as did both mixed tumors, one of the oncocytic adenomas and the papillary cystadenoma. One can conclude that the majority of sinonasal tract adenomatous neoplasms arise from the mucosal lining of the sinonasal tract. PMID- 2548184 TI - The mouse homolog of the hst/k-FGF gene is adjacent to int-2 and is activated by proviral insertion in some virally induced mammary tumors. AB - The fibroblast growth factor-related protooncogenes, int-2 and hst/k-FGF, are within 17 kilobase pairs of one another on mouse chromosome 7 and are in the same transcriptional orientation. Approximately 70% of tumors induced in BR6 mice by mouse mammary tumor virus have proviral insertions adjacent to the int-2 gene. We find that the murine homolog of the hst/k-FGF gene can also be transcriptionally activated by the insertion of mouse mammary tumor virus DNA either upstream or downstream of the gene. In most tumors, only one of these adjacent genes is activated, but in some cases both genes are expressed. One of the hst-expressing tumors also has a virally activated int-3 gene. At least five distinct cellular genes (int-1, -2, -3, -4, and hst/k-FGF) can therefore contribute, either singly or in concert, to the development of histologically indistinguishable mammary tumors in mice infected by mouse mammary tumor virus. PMID- 2548185 TI - Transmembrane signaling by a chimera of the Escherichia coli aspartate receptor and the human insulin receptor. AB - Since many receptors apparently contain only one or two membrane-spanning segments, their transmembrane topology should be similar. This feature suggests that these receptors share common mechanisms of transmembrane signaling. To test the degree of conservation of signaling properties, a chimeric receptor containing the ligand-binding extracellular domain of the Escherichia coli aspartate chemoreceptor and the cytosolic portion of the human insulin receptor was constructed. This chimeric receptor is active as a tyrosine kinase, and aspartate stimulates its activity. Some interesting differences are noted in the target proteins phosphorylated by the chimera compared to the wild-type insulin receptor. These results indicate that features of the signaling mechanisms used by these diverse receptors are conserved, but that interesting changes in the protein properties are caused by differences in the neighboring domains. PMID- 2548186 TI - Constitutive expression of c-myc oncogene confers hormone independence and enhanced growth-factor responsiveness on cells transformed by human papilloma virus type 16. AB - The effect of constitutive expression of the c-myc oncogene on the biological properties of cells transformed or immortalized by human papilloma virus type 16 (HPV16) was studied. Whereas transfection of HPV16 alone into primary baby mouse kidney (BMK) cells failed to generate any immortalized cell lines unless the tumor promoter phorbol 12-myristate 13-acetate was present, cotransfection of HPV16 with a plasmid that constitutively expresses murine c-myc (pSVc-myc-1) generated numerous rapidly growing colonies of cells. Cell lines transformed by HPV16 and pSVc-myc-1 did not require phorbol ester or steroid hormones for growth and were tumorigenic in syngeneic immunocompetent mice. Transfection of pSVc-myc 1 into established cell lines transformed by HPV16 and the v-fos oncogene increased the growth rate and saturation density of these lines severalfold, allowed growth in low-serum medium, and abolished the requirement of these cell lines for glucocorticoids or progestogens. Transfection of the EJ-ras oncogene into these lines did not significantly affect any of these properties. PMID- 2548187 TI - Actin polymerization induces a shape change in actin-containing vesicles. AB - We have encapsulated actin filaments in the presence and absence of various actin binding proteins into lipid vesicles. These vesicles are approximately the same size as animal cells and can be characterized by the same optical microscopic and mechanical techniques used to study cells. We demonstrate that the initially spherical vesicles can be forced into asymmetric, irregular shapes by polymerization of the actin that they contain. Deformation of the vesicles requires that the actin filaments be on average at least approximately 0.5 micron long as shown by the effects of gelsolin, an actin filament-nucleating protein. Filamin, a filament-crosslinking protein, caused the surfaces of the vesicles to have a smoother appearance. Heterogeneous distribution of actin filaments within the vesicles is caused by interfilament interactions and modulated by gelsolin and filamin. The vesicles provide a model system to study control of cell shape and cytoskeletal organization, membrane-cytoskeleton interactions, and cytomechanics. PMID- 2548188 TI - Phorbol 12-myristate 13-acetate inhibits binding of leukotriene B4 and platelet activating factor and the responses they induce in neutrophils: site of action. AB - The addition of the platelet-activating factor (PAF) to neutrophils causes an increase in cytoskeletal actin, a rise in the intracellular concentration of free calcium, release of arachidonic acid, and the synthesis of PAF. The PAF synthesis in human neutrophils stimulated by PAF is greatly potentiated by the human granulocyte-macrophage colony-stimulating factor. Incubation of human neutrophils with the tumor copromoter phorbol 12-myristate 13-acetate (PMA) for 3 min prior to the addition of the stimulus inhibits all these responses produced by PAF. The inhibition is prevented when the cells are incubated with protein kinase C inhibitors such as 1-(5-isoquinolinesulfonyl)-2-methylpiperazine for 5 min prior to the addition of PMA. The rise in the intracellular concentration of free calcium in human neutrophils stimulated with leukotriene B4 is also inhibited by PMA, and this inhibition is prevented by protein kinase C inhibitors such as staurosporine. Unlike PMA, the inactive ester 4 alpha-phorbol 12,13-didecanoate has no inhibitory effect on the stimulated rise in the intracellular concentration of free calcium. The binding of either PAF or leukotriene B4 to intact cells is inhibited by PMA. The most important finding of the present studies is that PMA interferes with the binding of PAF and leukotriene B4 to their respective receptors. Whether PMA inhibits the binding of these lipid mediators by activating protein kinase C or by perturbing the membrane directly remains to be elucidated. PMID- 2548189 TI - Translation of glucose-regulated protein 78/immunoglobulin heavy-chain binding protein mRNA is increased in poliovirus-infected cells at a time when cap dependent translation of cellular mRNAs is inhibited. AB - All cellular cytoplasmic mRNAs carry a 7-methylguanylate cap attached to their 5' ends. This cap structure is recognized by cap-binding proteins that then direct the binding of ribosomal subunits to this 5'-end complex. Poliovirus, a plus stranded RNA virus, interferes with this cellular translation process by proteolytically inactivating the cap-binding protein complex. Subsequently the viral mRNA can be translated by an initiation process in which ribosomes bind internally to the mRNA [Pelletier, J. & Sonenberg, N. (1988) Nature (London) 334, 320-325], obviating cap-dependent translation. At least one cellular mRNA, encoding a heat shock-like protein, glucose-regulated protein 78/immunoglobulin heavy-chain binding protein, has been discovered to be translated at an increased rate in poliovirus-infected cells at a time when the translation of other cellular mRNAs is inhibited. The glucose-regulated protein 78/immunoglobulin heavy-chain binding protein mRNA thus exemplifies a cellular mRNA that is translated at a specifically enhanced rate by an as-yet-unresolved cap independent initiation process in cells when the cap-binding protein complex is not functional. PMID- 2548190 TI - Identification and characterization of nuclear retinoic acid-binding activity in human myeloblastic leukemia HL-60 cells. AB - Specific [3H]retinoic acid (RA)-binding sites in nuclear and cytosolic extracts prepared from human myeloblastic leukemia HL-60 cells have been detected by sucrose density gradient sedimentation and size-exclusion high-performance liquid chromatography (HPLC) analyses. This RA-binding activity migrated as a single peak with an apparent molecular weight of 50,000 and greater than 95% of the total binding activity was associated with the nuclear extract. Nuclear extracts prepared from COS-1 cells transfected with an expression vector for the nuclear RA receptors RAR alpha or RAR beta were enriched (20- to 100-fold) with a RA binding activity that coeluted by size-exclusion HPLC with the putative RAR from HL-60 cells. The HL-60 nuclear receptor exhibited high-affinity binding of RA and its benzoic acid analogs Ch55, Ch30, Ro 13-7410, and SRI 6409-40 and low-affinity binding of retinol, Ro 8-8717, and SRI 5442-60, correlating well with the biological activity of these compounds in HL-60 cells. Saturation binding and Scatchard plot analyses of the binding of RA to the nuclear HL-60 receptor yielded an apparent dissociation constant of approximately 0.46 nM and 1400 +/- 100 receptor sites per cell. Northern blot analyses of poly(A)+ RNA with cDNA probes specific for RAR alpha and RAR beta indicated that HL-60 cells contain predominantly transcripts encoded by the RAR alpha gene. Our results suggest that the observed nuclear RA-binding activity in HL-60 cells might mediate the action of RA in these cells. PMID- 2548191 TI - In situ autoradiography and ligand-dependent tyrosine kinase activity reveal insulin receptors and insulin-like growth factor I receptors in prepancreatic chicken embryos. AB - We previously reported specific cross-linking of 125I-labeled insulin and 125I labeled insulin-like growth factor I (IGF-I) to the alpha subunit of their respective receptors in chicken embryos of 20 somites and older. To achieve adequate sensitivity and localize spatially the receptors in younger embryos, we adapted an autoradiographic technique using whole-mounted chicken blastoderms. Insulin receptors and IGF-I receptors were expressed and could be localized as early as gastrulation, before the first somite is formed. Relative density was analyzed by a computer-assisted image system, revealing overall slightly higher binding of IGF-I than of insulin. Structures rich in both types of receptors were predominantly of ectodermal origin: Hensen's node in gastrulating embryos and neural folds, neural tube and optic vesicles during neurulation. The signal transduction capability of the receptors in early organogenesis was assessed by their ability to phosphorylate the exogenous substrate poly(Glu80Tyr20). Ligand dependent tyrosine phosphorylation was demonstrable with both insulin and IGF-I in glycoprotein-enriched preparations from embryos at days 2 through 6 of embryogenesis. There was a developmentally regulated change in ligand-dependent tyrosine kinase activity, with a sharp increase from day 2 to day 4, in contrast with a small increase in the ligand binding. Binding of 125I-labeled IGF-I was, with the solubilized receptors, severalfold higher than binding of 125I-labeled insulin. However, the insulin-dependent phosphorylation was as high as the IGF-I dependent phosphorylation at each developmental stage. PMID- 2548192 TI - Tn5supF, a 264-base-pair transposon derived from Tn5 for insertion mutagenesis and sequencing DNAs cloned in phage lambda. AB - We constructed a derivative of transposon Tn5 called Tn5supF for insertion mutagenesis and sequencing DNAs cloned in phage lambda. This element carries a supF amber-suppressor tRNA gene. Its insertion into lambda can be selected by plaque formation by using nonsuppressing (sup0) Escherichia coli for amber mutant lambda phage and sup0 dnaB-amber E. coli for nonamber lambda phage. Tn5supF is just 264 base pairs long. It transposes efficiently and inserts quasi-randomly into DNA targets. The unique sequences near its termini can be used as primer binding sites for dideoxynucleotide DNA sequencing, thus permitting the direct sequencing of DNAs cloned in phage lambda without subcloning. PMID- 2548193 TI - Studies of the mechanisms of action of the antiretroviral agents hypericin and pseudohypericin. AB - Administration of the aromatic polycyclic dione compounds hypericin or pseudohypericin to experimental animals provides protection from disease induced by retroviruses that give rise to acute, as well as slowly progressive, diseases. For example, survival from Friend virus-induced leukemia is significantly prolonged by both compounds, with hypericin showing the greater potency. Viremia induced by LP-BM5 murine immunodeficiency virus is markedly suppressed after infrequent dosage of either substance. These compounds affect the retroviral infection and replication cycle at least at two different points: (i) Assembly or processing of intact virions from infected cells was shown to be affected by hypericin. Electron microscopy of hypericin-treated, virus-producing cells revealed the production of particles containing immature or abnormally assembled cores, suggesting the compounds may interfere with processing of gag-encoded precursor polyproteins. The released virions contain no detectable activity of reverse transcriptase. (ii) Hypericin and pseudohypericin also directly inactivate mature and properly assembled retroviruses as determined by assays for reverse transcriptase and infectivity. Accumulating data from our laboratories suggest that these compounds inhibit retroviruses by unconventional mechanisms and that the potential therapeutic value of hypericin and pseudohypericin should be explored in diseases such as AIDS. PMID- 2548194 TI - Neuromuscular recovery using calcium protease inhibition after median nerve repair in primates. AB - Inhibition of calcium-activated neutral protease, in muscle and nerve, by the tripeptide leupeptin after median nerve transection and epineural repair in monkeys (Cebus apella) was studied. Results indicate that inhibition of the protease after nerve repair facilitates morphologic recovery in denervated thenar muscles and in distal thenar nerve branches. In addition, functional recovery was facilitated in leupeptin-treated animals after nerve repair as measured by sensory and motor conduction velocities. Toxicologic testing showed that leupeptin, administered at 18 mg/kg, intramuscularly, twice daily, for 6 months did not adversely affect hematology, clotting, or plasma complement component C3 profiles. These data indicate that leupeptin is an effective and safe adjunct to peripheral nerve repair. PMID- 2548195 TI - Dynamic properties of the Ca2+/calmodulin-dependent protein kinase in Drosophila: identification of a synapsin I-like protein. AB - Visual adaptation with blue light induces a change in a special light/dark choice behavior in Drosophila. On the molecular level adaptation induces long-term modulation of the in vitro autophosphorylation capacity of a Ca2+/calmodulin dependent protein kinase. Here I describe a Drosophila phosphoprotein that is a substrate of this protein kinase. The molecular mass and phosphopeptide composition of this protein are similar to those of rat synapsin I. Furthermore, the Drosophila protein shows immunological cross-reactivity with monoclonal antibodies against rat synapsin I. I conclude that this 86-kDa protein in Drosophila is homologous to the vertebrate synapsin I. PMID- 2548196 TI - Recombination of knotted substrates by Tn3 resolvase. AB - We studied the site orientation specificity for recombination by purified Tn3 resolvase. With standard plasmid substrates, resolvase acts only on directly repeated recombination sites. Knotting, however, makes inverted site substrates equally efficient. The structure of the knotted products of recombination shows that the DNA wrapped around resolvase in the synaptic intermediate has the same local geometry as the standard substrate but is reversed in topological sign. Similarly, the same strand exchange with the two substrates generates supercoils with opposite signs. Thus, DNA geometry rather than topology is critical for these features of recombination. The knotted inverse substrate like the direct site substrate must be (-) supercoiled under standard reaction conditions. However, under conditions in which supercoiling is not required, the structure of the knotted product is apparently the same. This indicates that the unique direction of strand exchange is determined by the structure of the synaptosome and not by (-) supercoiling of the substrate. PMID- 2548197 TI - Stimulation of phosphorylation of lipocortin at threonine residues by epidermal growth factor (EGF) and the EGF receptor: addition of protein kinase P with polylysine inhibits this effect. AB - In this paper we show that epidermal growth factor (EGF) stimulates the phosphorylation of lipocortin 1, at threonine as well as at tyrosine residues, by a highly purified preparation of the EGF receptor. The phosphorylation of threonine residues is catalyzed by an enzyme that contaminates the receptor preparations, since crude extracts of A431 plasma membranes contain larger amounts of the threonine kinase than does the receptor preparation. Protein kinase P (2.5 ng) inhibits both threonine and tyrosine phosphorylation of lipocortin 1 while greatly stimulating the autophosphorylation of the EGF receptor. Acetyllipocortin 1 is poorly phosphorylated at tyrosine residues by the EGF receptor kinase, but it becomes readily phosphorylated in the presence of polylysine. The most likely explanation for this observation is that there is an interaction between polylysine and acetyllipocortin that converts the latter into a suitable substrate for the EGF receptor. These and other experiments described in this paper point to a role of surface charges in the susceptibility of substrates to attach by protein kinases. PMID- 2548198 TI - Dissection of functional domains of adenovirus DNA polymerase by linker-insertion mutagenesis. AB - Linker-insertion mutations were introduced into the cloned adenovirus DNA polymerase gene and the functional effects on the initiation and elongation of DNA in vitro were measured. Essential regions of the polymerase appear to be scattered in patches across the entire molecule and are not limited to the five regions of homology shared with a variety of other replicating polymerases. Thus, the adenovirus DNA polymerase presumably contains active sites that must be formed by distant interactions across the polymerase molecule. PMID- 2548200 TI - Cap-independent translation of mRNA conferred by encephalomyocarditis virus 5' sequence improves the performance of the vaccinia virus/bacteriophage T7 hybrid expression system. AB - A recombinant vaccinia virus that directs the synthesis of bacteriophage T7 RNA polymerase provides the basis for the expression of genes that are regulated by T7 promoters in mammalian cells. The T7 transcripts, which account for as much as 30% of the total cytoplasmic RNA at 24 hr after infection, are largely uncapped. To improve the translatability of the uncapped RNA, the encephalomyocarditis virus (EMCV) untranslated region (UTR) was inserted between the T7 promoter and the chloramphenicol acetyltransferase (CAT) gene. Experiments with a reticulocyte extract demonstrated that the EMCV UTR conferred efficient and cap-independent translatability to CAT RNA synthesized in vitro by T7 RNA polymerase. In cells infected with recombinant vaccinia viruses containing the T7 promoter-regulated CAT gene, the EMCV UTR increased the amount of CAT RNA on polyribosomes. The polyribosome-derived CAT RNA, which contained the EMCV UTR, was translated in vitro in a cap-independent fashion as well. Use of the EMCV UTR significantly enhanced the vaccinia/T7 hybrid expression system as it resulted in a 4- to 7 fold increase in total CAT activity. A further approximately 2-fold improvement was achieved by incubating the cells in hypertonic medium, which favors the translation of uncapped picornavirus RNA over cellular mRNAs. With this newly modified expression system, CAT was the predominant protein synthesized by infected cells and within 24 hr accounted for greater than 10% of the total cell protein. PMID- 2548201 TI - High-order fluorescence fluctuation analysis of model protein clusters. AB - The technique of high-order fluorescence fluctuation autocorrelation for detecting and characterizing protein oligomers was applied to solutions containing two fluorescent proteins in which the more fluorescent proteins were analogues for clusters of the less fluorescent ones. The results show that the model protein clusters can be detected for average numbers of observed subunits (free monomers plus monomers in oligomers) equal to 10-100 and for relative fluorescent yields that correspond to oligomers as small as trimers. High-order fluorescent fluctuation analysis may therefore be applicable to cell surface receptor clusters in natural or model membranes. PMID- 2548199 TI - Template supercoiling during ATP-dependent DNA helix tracking: studies with simian virus 40 large tumor antigen. AB - Incubation of topologically relaxed plasmid DNA with simian virus 40 (SV40) large tumor antigen (T antigen), ATP, and eubacterial DNA topoisomerase I resulted in the formation of highly positively supercoiled DNA. Eukaryotic DNA topoisomerase I could not substitute for eubacterial DNA topoisomerase 1 in this reaction. Furthermore, the addition of eukaryotic topoisomerase I to a preincubated reaction mixture containing both T antigen and eubacterial topoisomerase I caused rapid relaxation of the positively supercoiled DNA. These results suggest that SV40 T antigen can introduce topoisomerase-relaxable supercoils into DNA in a reaction coupled to ATP hydrolysis. We interpret the observed T antigen supercoiling reaction in terms of a recently proposed twin-supercoiled-domain model that describes the mechanics of DNA helix-tracking processes. According to this model positive and negative supercoils are generated ahead of and behind the moving SV40 T antigen, respectively. The preferential relaxation of negative supercoils by eubacterial DNA topoisomerase I explains the accumulation of positive supercoils in the DNA template. The supercoiling assay using DNA conformation-specific eubacterial DNA topoisomerase I may be of general use for the detection of ATP-dependent DNA helix-tracking proteins. PMID- 2548202 TI - Clonal heterogeneity of synovial fluid T lymphocytes from patients with rheumatoid arthritis. AB - Although substantial evidence suggests that synovial T lymphocytes are critical in the pathogenesis of rheumatoid arthritis (RA), little is known regarding their antigenic specificities, antigen receptor gene rearrangements, and mechanisms of activation. To assess the extent of expansion of specific clones among RA synovial fluid T cells, Southern blot analyses of T-cell receptor (TCR) gene rearrangements were performed on 40 RA synovial fluid T-cell clones, as well as on both fresh and polyclonally activated T cells from RA synovial fluid, RA peripheral blood, and normal peripheral blood. Two of the clones had identical TCR rearrangement patterns, but the remainder were unique. The nonclonal RA T cell samples showed the same pattern of TCR beta-chain rearrangement that was observed among normal peripheral blood T cells, indicating no dominant clonal T cell population in these samples. It was noted that with sufficient exposure of autoradiograms of the Southern blots, discrete TCR gene rearrangements, representing in some cases common D beta J beta (D, diversity; J, joining) rearrangements, were evident in T cells from peripheral blood of normal individuals and patients with RA, as well as T cells from RA synovial fluid. Taken together, the findings indicate that only a minor degree of oligoclonality can be demonstrated among T lymphocytes from RA synovial fluid. PMID- 2548203 TI - Internal frameshifts within the mitochondrial genes for cytochrome oxidase subunit II and maxicircle unidentified reading frame 3 of Leishmania tarentolae are corrected by RNA editing: evidence for translation of the edited cytochrome oxidase subunit II mRNA. AB - The Leishmania tarentolae cytochrome oxidase (EC 1.9.3.1) subunit II (COII) and maxicircle unidentified reading frame 3 (MURF3) mRNAs are edited internally by the addition of four and five uridine residues, respectively, which eliminate -1 and +1 reading frameshifts in the gene sequences. The editing events in COII are conserved in three kinetoplastid species, and those in MURF3 are conserved in two species. A primer extension assay showed that the ratio of edited to unedited RNA differed for each gene: 89% of the COII and 36% of the MURF3 transcripts are edited. Preliminary evidence was obtained for translation of the edited COII transcript into protein: antibodies generated against a synthetic peptide with the predicted carboxyl-terminal amino acid sequence reacted with a polypeptide of the correct molecular weight in immunoblots of a mitochondrial lysate. PMID- 2548204 TI - Rapid activation of the T-cell tyrosine protein kinase pp56lck by the CD45 phosphotyrosine phosphatase. AB - T lymphocytes express a tyrosine protein kinase (TPK; protein-tyrosine kinase; ATP:protein-tyrosine O-phosphotransferase, EC 2.7.1.112), pp56lck that is encoded by the lck protooncogene. This TPK was recently found to be associated with the intracellular domain of the T-cell surface glycoproteins, CD4 and CD8, suggesting that it plays an important role in T-cell development and activation. We have studied the regulation of pp56lck and found that this kinase can be rapidly activated by an endogenous mechanism present in T-lymphocyte membranes. This activation was sensitive to sodium orthovanadate and O-phosphotyrosine, consistent with the involvement of a phosphotyrosine phosphatase (PTPase; protein tyrosine-phosphatase; protein-tyrosine-phosphate phosphohydrolase, EC 3.1.3.48) in pp56lck activation. Based on a recent report demonstrating that CD45, the leukocyte common antigen, is a membrane-bound PTPase, we analyzed its role in pp56lck activation. CD45 was found to be the major (greater than 90%) PTPase in membranes of the murine T-lymphoma line BW5147. Moreover, activation of pp56lck was undetectable in a mutant BW5147 line lacking CD45 expression (and the associated PTPase activity). In contrast, activation of pp56lck was readily detected in the wild-type lymphoma line. More important, when immunoprecipitated CD45 was added to pp56lck, the TPK activity of the latter increased greater than 2-fold within minutes. This effect of CD45 was completely blocked by sodium orthovanadate. These findings indicate an important role for the CD45 PTPase in pp56lck activation. This role could be mediated by direct dephosphorylation of a regulatory tyrosine residue in pp56lck. PMID- 2548205 TI - Stimulatory effects of opioid neuropeptides on locomotory activity and conformational changes in invertebrate and human immunocytes: evidence for a subtype of delta receptor. AB - The presence of opioid neuropeptides was shown to stimulate conformational changes and locomotory activity in immunocytes of two representatives of invertebrates as well as in human leukocytes. Cells were examined by use of phase contrast and Nomarski optics coupled with a Zeiss Axiophot microscope, and of the Zeiss Videoplan/Vidas Image Analysis system. Immunocompetent blood cells, activated by exogenous opioids or stressful stimuli presumed to engage endogenous opioids, showed flattening, elongation, and formation of pseudopodia. In the mollusc Mytilus edulis, ameboid movements resulted in the formation of cell clusters, an activity not observed in untreated controls, or in immunocytes simultaneously exposed to opioid and naloxone. Tests with nine immunoreactive substances revealed immunocyte stimulation by delta, mu-, kappa-, and epsilon(?) selective ligands. One of these, [D-Ala2,D-Met5]enkephalinamide (DAMA), active at a concentration of 10 pM, proved to be considerably more effective than the rest. The high pharmacological potency of DAMA, observed in both human and invertebrate immunocytes, sets this opioid apart from the closely related [D-Ala2,D Leu5]enkephalin, a discrepancy not occurring in the mammalian nervous system. This suggests a specific function for [Met]enkephalin in immunoregulation, mediated perhaps by a special subtype of delta receptor. PMID- 2548206 TI - Highly potent metallopeptide analogues of luteinizing hormone-releasing hormone. AB - Metal complexes related to the cytotoxic complexes cisplatin [cis diamminedichloroplatinum(II)] and transbis(salicylaldoximato)copper(II) were incorporated into suitably modified luteinizing hormone-releasing hormone (LH-RH) analogues containing D-lysine at position 6. Some of the metallopeptides thus obtained proved to be highly active LH-RH agonists or antagonists. For instance, SB-40, a PtCl2-containing metallopeptide in which platinum is coordinated to an N epsilon-(DL-2,3-diaminopropionyl)-D-lysine residue [D-Lys(DL-A2pr] at position 6, showed 50 times higher LH-releasing potency than the native hormone. SB-95, [Ac-D Nal(2)1,D-Phe(pCl)2, D-Pal(3)2, Arg5,D-Lys[DL-A2pr(Sal2Cu)]6,D-Ala10]LH-RH, where Nal(2) is 3-(2-naphthyl)alanine, Pal(3) is 3-(3-pyridyl)alanine, and copper(II) is coordinated to the salicylideneimino moieties resulting from condensation of salicylaldehyde with D-Lys(DL-A2pr)6, caused 100% inhibition of ovulation at a dose of 3 micrograms in rats. Most metallopeptide analogues of LH-RH showed high affinities for the membrane receptors of rat pituitary and human breast cancer cells. Some of these metallopeptides had cytotoxic activity against human breast cancer and prostate cancer cell lines in vitro (this will be the subject of a separate paper on cytotoxicity evaluation). Such cytostatic metallopeptides could be envisioned as targeted chemotherapeutic agents in cancers that contain receptors for LH-RH-like peptides. PMID- 2548207 TI - Highly potent analogues of luteinizing hormone-releasing hormone containing D phenylalanine nitrogen mustard in position 6. AB - The nitrogen mustard derivatives of 4-phenylbutyric acid and L-phenylalanine, called chlorambucil (Chl) and melphalan (Mel), respectively, have been incorporated into several peptide hormones, including luteinizing hormone releasing hormone (LH-RH). The alkylating analogues of LH-RH were prepared by linking Chl, as an N-acyl moiety, to the complete amino acid sequence of agonistic and antagonistic analogues. These compounds, in particular the antagonistic analogues, showed much lower potency than their congeners carrying other acyl groups. To obtain highly potent alkylating analogues of LH-RH, the D enantiomer of Mel was incorporated into position 6 of the native hormone and some of its antagonistic analogues. Of the peptides prepared, [D-Mel6]LH-RH (SB-05) and [Ac-D-Nal(2)1,D-Phe(pCl)2,D-Pal(3)3,Arg5,D-Mel6,D-Ala10++ +]LH-RH [SB-86, where Nal(2) is 3-(2-naphthyl)alanine and Pal(3) is 3-(3-pyridyl)alanine] possessed the expected high agonistic and antagonistic activities, respectively, and also showed high affinities for the membrane receptors of rat pituitary cells, human breast cancer cells, human prostate cancer cells, and rat Dunning R 3327 prostate tumor cells. These two analogues exerted cytotoxic effects on human and rat mammary cancer cells in vitro. Thus these two D-Mel6 analogues seem to be particularly suitable for the study of how alkylating analogues of LH-RH could interfere with intracellular events in certain cancer cells. PMID- 2548208 TI - Human neutrophils and mononuclear cells inhibit platelet aggregation by releasing a nitric oxide-like factor. AB - Incubation of neutrophils or mononuclear cells with washed platelets (all prepared from human venous blood) resulted in an inhibition of thrombin-induced platelet aggregation that was dependent on the number of nucleated cells added. The inhibition was potentiated by superoxide dismutase and reversed by oxyhemoglobin. In the case of neutrophils the inhibition was associated with an increase in cGMP, whereas with mononuclear cells both cAMP and cGMP were increased. The inhibitory activity of neutrophils or mononuclear cells was prevented by their preincubation with NG-monomethyl-L-arginine methyl ester. L Arginine reversed the action of NG-monomethyl-L-arginine methyl ester, whereas D arginine was ineffective. Preincubation of the cells with catalase or mannitol did not prevent their inhibitory action on platelet aggregation. The inhibition of platelet aggregation was not due to platelet damage or to uptake of thrombin by neutrophils or mononuclear cells. It was overcome by increasing the concentration of thrombin and was absent in cell-free supernatants obtained from a suspension of neutrophils or mononuclear cells or from mixtures of platelets with neutrophils or platelets with mononuclear cells. These data provide evidence for the release of a nitric oxide-like factor from human neutrophils and mononuclear cells. In addition, evidence is provided that, as in stimulated murine macrophages and endothelial cells, the precursor of this factor is L arginine. PMID- 2548209 TI - Epstein-Barr virus-transformed pro-B cells are prone to illegitimate recombination between the switch region of the mu chain gene and other chromosomes. AB - Six independently maintained sublines of FLEB 14, a fetal-liver-derived Epstein Barr virus-transformed pro-B cell line that has not yet rearranged its immunoglobulin genes, were examined after in vitro propagation during 19-36 months. Two lines showed no immunoglobulin heavy chain gene rearrangement, whereas one allele was rearranged with breakpoints inside the switch region of the mu chain gene in the remaining four. These rearrangements had been generated by the translocation of different chromosome fragments to the immunoglobulin heavy chain gene cluster-carrying 14q32 band in each of the four lines. Previously, a similar rearrangement was found in a fifth subline concurrently with a reciprocal 6;14 translocation. The transposed pieces have been derived from chromosomes 16 and 18 in two of the more recently rearranged lines. Their origins could not be determined in the remaining two lines, but they were different from each other and the other three 14q+ markers. The 14q+ marker carrying variant has replaced its diploid progenitor suggesting that the translocation has conveyed some in vitro growth advantage on its carrier. This was also supported by the duplication of the 14q+ marker and the loss of its normal chromosome 14 homologue in one subline during serial culturing. The vulnerability of the switch region of the mu chain gene to illegitimate recombination at the pro-B stage and the possible relevance of this finding for the origin of the Burkitt lymphoma-associated 8;14 (immunoglobulin heavy chain gene cluster/MYC) translocation is discussed. PMID- 2548210 TI - Vaccine protection against simian immunodeficiency virus infection. AB - Rhesus monkeys were immunized by multiple inoculations with purified, disrupted, noninfectious simian immunodeficiency virus (SIV) in adjuvant. Immunized monkeys developed anti-SIV antibodies detectable by whole-virus ELISA and by immunoblot reactivity; these antibodies had weak neutralizing activity. One week after the last immunization, monkeys were challenged with 200-1000 animal infectious doses of uncloned, live SIV. The same strain of SIV that was used for vaccination was also used for challenge. Anamnestic antibody responses and SIV recovery from peripheral blood were used to evaluate infection following the live virus challenge; two of six vaccinated monkeys showed no evidence of infection following the live virus challenge. Transfusion of 10 ml of whole blood from these two into uninfected, naive rhesus monkeys did not result infection of the recipients, providing further support for the lack of infection in the two previously vaccinated animals. Four of four unvaccinated control monkeys inoculated with these doses of live SIV became infected and three of these died with AIDS 118-258 days after infection. Only one of the six vaccinated monkeys has died to date. In situ hybridization with lymph node biopsy specimens suggested that the virus load was much higher in control macaques than in vaccinated macaques. These results indicate that vaccination with inactivated whole virus can protect macaques against challenge with live SIV. Furthermore, they provide hope that vaccine protection against human AIDS virus infection may be possible. PMID- 2548211 TI - Cloning and molecular characterization of genes whose products allow Salmonella typhimurium to penetrate tissue culture cells. AB - Invasion of the intestinal epithelium is thought to be an important step in the pathogenesis of Salmonella infections. Using an in vitro system, we have isolated a genetic locus, inv, that confers to a noninvasive strain of Salmonella typhimurium the ability to penetrate tissue culture cells. Highly virulent S. typhimurium strains carrying inv mutations were defective for entry into Henle 407 cells while remaining unaffected in their ability to attach to cultured cells. When administered perorally to BALB/c mice, inv mutants of S. typhimurium had higher 50% lethal doses (LD50) than their wild-type parent strains. To the contrary, there were no differences in the observed LD50 when strains were administered intraperitoneally. In addition, inv mutants presented decreased ability to colonize the Peyer's patches, the small intestinal wall, and the spleen when administered perorally, although when administered intraperitoneally, they showed no difference in their ability to colonize the spleen compared to the wild-type parent strain. PMID- 2548212 TI - Selective changes in mu opioid receptor properties induced by chronic morphine exposure. AB - Chronic infusion of morphine to guinea pigs produced selective changes in mu agonist binding properties in cerebrocortical membrane preparations. Employing the mu-selective opioid agonist [D-Ala2,MePhe4,Gly-ol5]enkephalin (DAMGO) in direct binding studies and in competition of labeled antagonist binding, we found that the major changes were a decrease in the number of sites with high affinity for agonist, a small reduction in total receptor number, and a loss in the ability of guanosine 5'-[gamma-thio]triphosphate to regulate binding. A fraction of high-affinity mu receptors appeared to retain their high affinity for agonist and their sensitivity to guanine nucleotide analogue after the induction of morphine tolerance, possibly because the morphine concentrations achieved in brain were insufficient to uncouple all mu receptors from associated guanine nucleotide-binding regulatory proteins. Some membrane preparations were treated with pertussis toxin, which has been shown to functionally uncouple mu opioid receptors from their effector systems. In these preparations, a single agonist affinity state of the receptor was observed. The apparent dissociation constant for this affinity state in pertussis toxin-treated membranes was similar to the lower-affinity state observed in preparations from morphine-tolerant animals. In contrast to the changes observed at mu opioid binding sites, no significant changes in agonist affinity or binding density were observed for selective delta or kappa agonists, consistent with the development of selective tolerance at mu receptors. PMID- 2548213 TI - Intracerebroventricular infusion of interleukin 1 rapidly decreases peripheral cellular immune responses. AB - Low doses (50-200 pg or 3.1-12.4 fmol) of interleukin 1 (IL-1) infused into the brain of rats produced rapid suppression of various cellular immune responses in peripheral lymphocytes of rats. Fifteen minutes after infusion of purified IL-1 beta into the lateral ventricle, natural killer cell activity, response to phytohemagglutinin stimulation, and interleukin 2 production were markedly suppressed in lymphocytes isolated from blood and spleen. These effects were due to infusion of IL-1 into brain since they did not occur when IL-1 was infused into the cisterna magna (essentially posterior to brain) or was injected intraperitoneally. Effects of IL-1 in brain could be blocked by simultaneous infusion of alpha-melanocyte-stimulating hormone, which is known to block the biological actions of IL-1. To stimulate release of endogenous IL-1 in brain, lipopolysaccharide was infused; this produced similar effects as IL-1, and these effects also were blocked by alpha-melanocyte-stimulating hormone. At longer intervals after infusion of IL-1 and lipopolysaccharide (3, 6, and 24 hr), immune responses returned to baseline or remained suppressed; i.e., "rebound" immunopotentiation did not occur. Finally, IL-1 infusion suppressed cellular immune responses in adrenalectomized animals, thereby showing that the effects of central IL-1 on peripheral cellular immune responses were, at least in part, independent of the stimulatory effect of IL-1 on secretion of adrenal hormones. These results indicate a link from brain to peripheral immune responses by means of action of a cytokine acting in the brain. PMID- 2548214 TI - Dopamine D2 receptors in the cerebral cortex: distribution and pharmacological characterization with [3H]raclopride. AB - An apparent involvement of dopamine in the regulation of cognitive functions and the recognition of a widespread dopaminergic innervation of the cortex have focused attention on the identity of cortical dopamine receptors. However, only the presence and distribution of dopamine D1 receptors in the cortex have been well documented. Comparable information on cortical D2 sites is lacking. We report here the results of binding studies in the cortex and neostriatum of rat and monkey using the D2 selective antagonist [3H]raclopride. In both structures [3H]raclopride bound in a sodium-dependent and saturable manner to a single population of sites with pharmacological profiles of dopamine D2 receptors. D2 sites were present in all regions of the cortex, although their density was much lower than in the neostriatum. The density of these sites in both monkey and, to a lesser extent, rat cortex displayed a rostral-caudal gradient with highest concentrations in the prefrontal and lowest concentrations in the occipital cortex, corresponding to dopamine levels in these areas. Thus, the present study establishes the presence and widespread distribution of dopamine D2 receptors in the cortex. PMID- 2548216 TI - Coordinated regulation of intracellular K+ in the proximal tubule: Ba2+ blockade down-regulates the Na+,K+-ATPase and up-regulates two K+ permeability pathways. AB - To avoid large changes in cell K+ content and volume during variations in Na+,K+ ATPase activity, Na+-transporting epithelia must adjust the rate of K+ exit through passive permeability pathways. Recent studies have shown that a variety of passive K+ transport mechanisms may coexist within a cell and may be functionally linked to the activity of the Na+,K+-ATPase. In this study, we have identified three distinct pathways for passive K+ transport that act in concert with the Na+,K+-ATPase to maintain intracellular K+ homeostasis in the proximal tubule. Under control conditions, the total K+ leak of the tubules consisted of discrete Ba2+-sensitive (approximately 65%), quinine-sensitive (approximately 20%), and furosemide-sensitive (approximately 10%) pathways. Following inhibition of the principal K+ leak pathway with Ba2+, the tubules adaptively restored cell K+ content to normal levels. This recovery of cell K+ content was inhibited, in an additive manner, by quinine and furosemide. Following adaptation to Ba2+, the tubules exhibited a 30% reduction in Na+-K+ pump rate coupled with an increase in K+ leak by means of the quinine-sensitive (approximately 70%) and furosemide sensitive (approximately 280%) pathways. Thus, the proximal tubule maintains intracellular K+ homeostasis by the coordinated modulation of multiple K+ transport pathways. Furthermore, these results suggest that, like Ba2+, other inhibitors of K+ conductance will cause compensatory changes in both the Na+-K+ pump and alternative pathways for passive K+ transport. PMID- 2548215 TI - Role of Ca2+ channels in the ability of membrane depolarization to prevent neuronal death induced by trophic-factor deprivation: evidence that levels of internal Ca2+ determine nerve growth factor dependence of sympathetic ganglion cells. AB - Sympathetic neurons depend on nerve growth factor (NGF) for their survival both in vivo and in vitro; these cells die upon acute deprivation of NGF. We studied the effects of agents that cause membrane depolarization on neuronal survival after NGF deprivation. High-K+ medium (greater than or equal to 33 mM) prevented cell death; the effect of K+ was dose-dependent (EC50 = 21 mM). The protection by high K+ was abolished either by withdrawal of extracellular Ca2+ or by preloading the cells with a Ca2+ chelator. The involvement of Ca2+ flux across membranes in high-K+ saving of NGF-deprived neurons was also supported by experiments using Ca2+-channel antagonists and agonists. The Ca2+ antagonists nimodipine and nifedipine effectively blocked the survival-promoting effect of high K+. The Ca2+ agonists Bay K 8644 and (S)-202-791 did not by themselves save neurons from NGF deprivation but did strongly augment the effect of high K+; EC50 was shifted from 21 mM to 13 mM. These data suggest that dihydropyridine-sensitive L-type Ca2+ channels play a major role in the high-K+ saving. The depolarizing agents choline (EC50 = 1 mM) and carbamoylcholine (EC50 = 1 microM), acting through nicotinic cholinergic receptors, also rescued NGF-deprived neurons. The saving effect of nicotinic agonists was not blocked by withdrawal of extracellular Ca2+ but was counteracted by a chelator of intracellular Ca2+, suggesting the possible involvement of Ca2+ release from internal stores. Based on these findings we propose a "Ca2+ set-point hypothesis" for the degree of trophic-factor dependence of sympathetic neurons in vitro. PMID- 2548217 TI - Electric field stimulation alters the outputs of prostaglandins from isolated rat urinary bladder preparations. Influences of papaverine and tetradotoxin. AB - The effects of electric field stimulation (EFS) on the outputs of prostaglandins (PGs) E1, E2 and F2 alpha from isolated contracting rat urinary bladders, were explored. Also, the influences of papaverine (5.10(-6) M) and of tetradotoxin (TTX = 5.10(-7) M) on PGs released by spontaneously contracting or electrically driven preparations, were tested. The basal control outputs of the three PGs in spontaneously contracting urinary bladders, had a comparable magnitude. On the contrary, in electrically stimulated preparations the output of PGE2 rose significantly; that of PGF2 alpha presented a significant reduction and the release of PGE1 was similar to that in controls. Papaverine failed to modify the profile of basal control PG release in non-stimulated bladders, abolished contractile responses to EFS and blocked the augmented output of PGE2 elicited by EFS. The presence of TTX in the suspending solution had no action on the basal control release of PGs from non-stimulated spontaneously contracting preparations, depressed between 80-90% the inotropic responses triggered by EFS and completely antagonized the enhanced output of PGE2 evoked by EFS. Results are discussed in terms of the relative participations of the evoked inotropism of the detrusor muscle or by the stimulation of nerve endings, accounting for the greater release of tissue PGE2 after EFS. PMID- 2548219 TI - Effect of oral ingestion of eicosapentaenoic acid-ethyl ester on natural killer cell activity in rat spleen cells. AB - Effect of ingestion of eicosapentaenoic acid-ethyl ester (EPA-E) on natural killer (NK) cell activity in rat spleen was examined. EPA-E ingestion during four weeks (100 mg/kg/day) significantly depressed NK cell activity. After EPA-E ingestion, EPA content in the spleen cell lipids was significantly increased, compared to that in control rat spleen cells, and AA content was not changed. Production of leukotriene B4 (LTB4) in spleen cells obtained from rats fed with EPA-E (EPA rats) was significantly decreased, compared to that from control rats. A decreased NK cell activity in spleen cells obtained from EPA rats was recovered by the addition of LTB4 in a dose-dependent manner. The current data indicate that EPA-E-induced inhibition of NK cell activity is mediated at least partially by the decrease of LTB4 production. PMID- 2548218 TI - Release of eicosanoids in rat peritoneal cavity stimulated with platelet activating factor (PAF). Effect of the PAF-antagonist BN-52021. AB - Platelet activating factor (PAF; 10 micrograms) was injected in the peritoneal cavity of rats in the absence or presence of the PAF antagonist BN-52021 (5 mg/kg). Thirty min later, the peritoneal cavity was washed with 3 ml of saline, the fluid was collected and the concentrations of selected eicosanoids were measured using novel enzyme immunoassays. PAF increased by 2.9, 2.8 and 1.7 fold the levels of thromboxane B2, prostaglandin E2 and leukotriene B4 respectively in the peritoneal fluid. The stimulatory effects of PAF was reduced by 42, 51, and 86% for thromboxane B2, prostaglandin E2 and leukotriene B4 respectively by the specific PAF antagonist. These results confirm the presence of specific PAF receptors in tissues and/or cells of rat peritoneal cavity and underline the complex interactions between PAF and eicosanoids. PMID- 2548221 TI - Recovery of viral capacity in irradiated exponentially growing cells. AB - Exponentially growing cells of the PtK-2 line (ATCC No. CCL56, from the marsupial Potorous tridactylus) require protein and RNA synthesis in a limited period following UV-radiation damage for optimal recovery as colony formers [Overberg et al. (1988) Mutat. Res. 194, 83-92]. Overall behavior suggests the operation of damage-induced recovery processes. The capacity of confluent cell monolayers for infection with unirradiated herpes simplex virus 1 (HSV-1) is sharply reduced by UV-irradiation. We have followed capacity changes in exponentially growing cells after irradiation and varying amounts of photoreactivation by means of an infectious center assay. These changes closely parallel changes of colony formation. Spontaneous recovery of capacity in the dark occurs over approximately the same time period that the UV sensitivity of colony formation depends on macromolecular synthesis. The effect of photoreactivation is complementary rather than additive to this recovery, suggesting that the dark recovery in this period concerns pyrimidine dimers in cell DNA. PMID- 2548220 TI - Chemotactic factor generation and cell accumulation in acute lung injury induced by endotracheal acid instillation. AB - We studied the time course of chemotactic factor generation and inflammatory cell accumulation in the rabbit aspiration pneumonia model. Two major potent chemotactic factors, leukotriene B4 (LTB4) and C5a, in bronchoalveolar lavage fluid (BALF) were measured by radioimmunoassay, and cell analysis was also done. The level of LTB4 increased only in the early phase (2-6 h) after endotracheal acid instillation. The level of C5a increased gradually almost in parallel with the total protein level in BALF, and reached a maximum at 24 h. Neutrophil accumulation occurred early and reached a maximum at 24 h. In contrast, the number of alveolar macrophages increased from days 1 to 7. These findings suggest that the increases in LTB4 and C5a are responsible for accumulation of neutrophils and that C5a may be an important chemotactic factor for alveolar macrophage. PMID- 2548222 TI - The effect of clonidine on auditory P300. AB - Auditory evoked potential recordings were done on 14 normal subjects during baseline conditions as well as after oral administration of 0.2 mg of clonidine or placebo. P300 amplitude and latency measurements were obtained from 28 electrodes and analyzed. Clonidine resulted in a decrease in P300 amplitude, which was most marked in the occipital and left parieto-temporal regions. This effect was significant even after controlling for the sedative side effect of clonidine. P300 was unaffected by clonidine. These data suggest the possible use of clonidine-induced changes in P300 amplitude as an index of central noradrenergic responsiveness. PMID- 2548223 TI - Platelet [3H]-imipramine binding is not modified in Alzheimer's disease. AB - Platelet [3H]-imipramine binding was studied in patients with Alzheimer's disease and control subjects matched to the patients for age and sex. There were no differences in the binding parameters of [3H]-imipramine on platelet membranes from patients with Alzheimer's disease, when compared with the control group. These results suggest that [3H]-imipramine binding could be a useful tool to discriminate between demented and depressive patients in elderly populations. PMID- 2548224 TI - Hypothalamo-pituitary-adrenal axis abnormalities in depression: a review and a model. AB - A wide range of abnormalities of the hypothalamo-pituitary-adrenal (HPA) axis has been described in depression. This paper reviews recent advances in the understanding of this system, and draws them together to construct a model for the purposes of further research and discussion. It is proposed that there are two fundamental changes which both originate in the hypothalamus: an increased secretion of corticotropin-releasing hormone, and a neurally mediated adrenal hyper-responsivity to ACTH. The resulting changes in hormone regulation would be expected to produce all the characteristic HPA axis abnormalities commonly seen in depression. The model makes several predictions which could be tested by future experiment. PMID- 2548225 TI - [The possible nature and species specificity of a protein factor enabling the solubilization of chromatin from thymocytes of irradiated animals]. PMID- 2548226 TI - [Effect of ionizing radiation at high doses on the cyclic nucleotide content of the rat brain]. AB - The study of the rat brain after irradiation with superlethal doses of 100 and 500 Gy has revealed a decrease in the cAMP content at early times after exposure. The most considerable decline in the cGMP content was noted in the cerebellum and cerebral hemisphere cortex where low cGMP level was retained for 24 h following irradiation. It is suggested that the observed changes play an important role in the development of the CNS syndrome. PMID- 2548227 TI - [The action of ionizing radiation on neuronal function in the edible snail. Phospholipase A2 and the Na+ and K+ transport systems]. AB - A study was made of the influence of A2 phospholipase on 22Na release from cells of nerve ganglia of edible snail. The treatment of nerve ganglia with A2 phospholipase inhibits Na, K-pump of neuronal membranes and does not exert a substantial effects on Na/Ca metabolism. There is a similarity between the effects of ionizing radiation and A2 phospholipase on the release of 22Na from cells. PMID- 2548229 TI - Opioids and respiration. PMID- 2548228 TI - [The action of gamma irradiation on trypsin immobilized on a modified polypropylene textile material]. AB - In studying the proteolytic activity and ESR spectra of gamma-irradiated samples of trypsin immobilized at an inoculated copolymer of polypropylene with polyacrylic acid it was established that the carrier of a modified polypropylene increases the radioresistance of trypsin immobilized on it. PMID- 2548231 TI - The biotransformation in vitro of cysteinyl leukotrienes in blood of normal and asthmatic subjects. AB - The metabolism of exogenous leukotriene C4 (LTC4), LTD4 and LTE4 (10(-8) M) was studied in vitro in blood of normal and asthmatic subjects for up to 2 hr by reverse-phase high performance liquid chromatography. In whole blood, incubation of LTC4 (T1/2 = 11.5 min) resulted in the formation of LTD4 and LTE4 whose biosynthesis was inhibited by serine borate (30 mM). Similar experiments performed with LTD4 (T1/2 = 5 min) produced a single metabolite (LTE4) which was inhibited by L-cysteine (10 mM). On the other hand, LTE4 represented a highly stable product in our in vitro system. The bioconversion of LTC4 or LTD4 was slower in plasma but this effect appeared more pronounced for the cysteinylglycinyl derivative. The bioconversion of LTD4 in whole blood or plasma was almost twice as rapid as LTC4. Experiments performed with asthmatic blood showed no significant difference in the survival of LTC4. These results suggest that blood may play a role in regulating the bioavailability of cysteinyl containing LTs which could be of relevance to their excretion in man. PMID- 2548230 TI - ACTH modulation of nerve development and regeneration. AB - (1) The availability of short amino acid sequences of the naturally occurring ACTH 1-39 molecule has made it possible to separate the corticotropic characteristics of the parent molecule from its neurotrophic effects. Potent neurotrophic fragments are ACTH 4-10, an analog of ACTH 4-9 (Org 2766), and alpha MSH (ACTH 1-13), peptide fragments that do not evoke corticosteroid secretion, yet clearly affect both the development and regeneration of peripheral nerve. (2) Early postnatal administration of either ACTH 4-10 or Org 2766 accelerates the neuromuscular development of the immature rat, increasing the contractile strength of the EDL muscle and inducing more rapid muscle contractions. Grasping strength and motor activity are increased; these are all changes indicative of more rapid neuromuscular maturation. Prenatal peptide treatment elicits a more complex pattern of response since administration early in gestation (GD 3-12) accelerates neuromuscular development whereas later administration (GD 13-21) decelerates maturation. (3) ACTH peptides have a similar accelerating effect on the morphology of the developing neuromuscular junction. At two weeks of age, nerve arborization is conspicuously increased by postnatal administration of either ACTH 4-10 or Org 2766, as is nerve terminal branching within the endplate itself. However, this is preceded by an initial depression of nerve branching in the 7-day-old rat pup. We conclude that while the developing neuromuscular system is sensitive to ACTH peptides, this susceptibility is age-related. The crucial role of these peptides may be limited to very brief, defined periods during which the peptides may interact with trophic or growth-associated substances, each of which may have its own decisive, circumscribed time frame of influence. (4) Perinatal administration of ACTH peptides affects CNS development. One measurable indication of this is an acceleration of eye opening. Early exposure to ACTH peptides has long-lasting effects on behavior, apparent when these animals are tested as adults. Increased spontaneous motor activity, heightened states of arousal and agitation, and changes in social behavior have been reported. Certain avoidance responses and tests of visual discrimination in male rats are improved by neonatal treatment with alpha-MSH. Overall motor activity is increased and the normal period of hyperactivity is initiated earlier. Male sexual behavior is decreased and sexually dimorphic behaviors in males are eliminated. alpha-MSH may alter the development of its own dopaminergic feedback circuitry while ACTH affects serotonin levels in the preoptic nucleus.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2548232 TI - Acute skin reactions observed in fractionated proton irradiation. AB - Between May 1985 and July 1987, 49 skin reactions of 43 patients treated by proton irradiation were observed at the Particle Radiation Medical Science Center (PARMS), The University of Tsukuba. Taking the peak skin score as an endpoint, the radiobiological effects [relative biological effectiveness (RBE) and time dose relationships] of the proton beam in multi-fractionated treatments were estimated. Factors influencing the skin dose, such as the prescribed tumor dose, tumor site, and number of applied fields, were also analyzed. The following conclusions regarding acute skin reactions to the clinical use of proton irradiation were obtained: 1) the physical skin-sparing effect of proton irradiation in single-field irradiation, especially in superficial regions, is not large compared with that of high-energy photon irradiation; 2) multidirectional proton irradiation significantly reduced the skin dose and severity of acute reactions; 3) the radiobiological effects of the proton beam, RBE and the time factor, estimated in human skin in multi-fractional treatment were slightly smaller than those of X-rays, i.e., 0.92 and -0.25 +/- 0.09, respectively. PMID- 2548233 TI - Arteriovenous malformation of the pancreas associated with mesenteric varices: case report and review of the literature. AB - A 53-year-old man with an arteriovenous malformation of the pancreas associated with extensive mesenteric varices, liver cirrhosis, and hepatocellular carcinoma with arterioportal shunting was diagnosed by angiography. This is the first report of such a case with portal hypertension. PMID- 2548234 TI - Optimization of proton and heavy ion therapy using an adaptive inversion algorithm. PMID- 2548235 TI - [Diagnostic, staging and therapeutic problems in invasion of the abdominal vena cava in renal cancer]. AB - Between 1976 and 1987 the authors have operated 351 renal carcinomas of the clear cell type, and 51 Wilms tumours. In 35 of the patients there was malignant thrombosis of the abdominal vena cava. Total resection of the vena cava was done in the subhepatic portion in 6 of the patients, and lateral resection with removal of the thrombus was carried out in another 15 patients. In the other 14 cases thrombosis of the vena cava was accompanied by visceral, lymph node metastases, or by invasion of the surrounding organs, and palliative nephrectomy alone was done in 13 patients. One of the patients declined surgery. A new staging is suggested, of the venous invasion, which correlates satisfactorily with the patients' perspectives. Invasion of the main renal venous pathway is noted by V1, invasion of the abdominal vena cava is noted by V2 (with subgroups V2a, when the thrombus is free in the vein, without contacts with the venous wall, and V2b, when the thrombus infiltrates the wall of the vena cava. In cases staged as N0M0V2a perifascial nephrectomy and unblocking of the abdominal vena cava may achieve remarkably long survival, while the V2b substage should be assimilated to visceral metastases. PMID- 2548236 TI - [Digital image intensifier radiography and film-screen combinations in pulmonary nodules]. AB - 30 patients with known or suspected pulmonary nodules were examined using a digital image intensifier radiography system, a conventional radiography system and a tomography of the whole lung. 73.8% of the pulmonary nodules detected with tomography were also detected with the digital system, 69.1% with the conventional system. There was no significant difference between the two systems comparing false positive and false negative diagnoses. Regarding the dose reduction to nearly 25% of that of a conventional examination the digital technique should be preferred to the conventional technique, if pulmonary nodules are suspected. PMID- 2548237 TI - [The clinical value of computed tomography in the pretherapeutic T-staging of orofacial tumors]. AB - The clinical role of contrast-enhanced CT for pre-operative T-staging of oro facial tumours was evaluated in 174 patients. During stages T1 and T2, clinical examination is 90% accurate and has a significantly higher sensitivity than CT, with an accuracy of only 53%. The clinical diagnosis of T1 and T2 is, however, made with undue frequency; CT makes it possible to obtain correct staging. Combination of CT and clinical examination can improve correct staging from 46% to 81%. During stages T3 and T4. CT has sensitivity of 91% and is markedly superior to clinical examination, with a sensitivity of 34%. Combined clinical and CT examination improves the predictive value from 52% to 92%. Combination of clinical and CT staging, when the higher stage is accepted as being correct, has a sensitivity and reliability of over 90%. It can be used as a firm basis for treatment planning, judging the prognosis and the effects of treatment. PMID- 2548239 TI - [Magnetic resonance tomographic studies of pancreatic transplants]. AB - In a pilot study, a total of 16 recipients of a combined pancreatic-renal allograft were examined by means of magnetic resonance imaging. Two groups were formed according to the management of the exocrine secretion: In group I patients (n = 8) the pancreatic duct was occluded with an alcoholic prolamine solution, in group II (n = 8) pancreatic juice was diverted either into a Roux-loop of jejunum (IIa, n = 4) or into the urinary bladder (group IIb, n = 4). On the T1-weighted images no difference was found between both groups with respect to signal intensity. In contrast, on T2-weighted images of group IIb, which were in situ for a significantly shorter time, a higher signal intensity was noticed as compared with the other patients. From these findings it is concluded that graft fibrosis may occur even in pancreas transplants with diversion of the exocrine secretion. PMID- 2548238 TI - [Ultrasonic diagnosis following liver transplantation]. AB - Realtime and duplex sonography were used routinely for the postoperative and long term follow-up of liver transplantation patients. In the early postoperative period 90 patients exhibited diffuse (67.7%) and focal (33.3%) alterations of the liver parenchyma; in 53% of patients various liquid lesions were diagnosed in the abdominal cavity. In case of diffuse alterations of the liver parenchyma, no correlation could be established between sonomorphology and histopathologic findings; especially rejection could not be diagnosed by using sonography. In case of focal intrahepatic lesions highly echogenic areas of infarction could be differentiated from poorly echogenic changes such as necrosis, haematoma and abscess--these entities had to be further distinguished by means of CT or needle biopsy. 20 patients showed biliary pathology (biliomas: 5, cholangiectasis: 15). In 73.3% of cholangiectasis the cause of obstruction could be demonstrated sonographically (biliary thrombus, stenosis of the anastomosis of the common bile duct, recurrence of tumour). In 13 patients major vascular complications were diagnosed predominantly by means of duplex sonography (hepatic artery occlusion: 4, hepatic artery stenosis: 1, portal vein occlusion: 1, partial thrombosis of portal vein: 2, portal vein stenosis: 2, stenosis with incomplete thrombosis of the inferior vena cava: 3, VCI stenosis: 1). 13 patients developed recurrent malignancy in the late postoperative period. PMID- 2548241 TI - [Quantitative computed tomographic flow measurements in aortocoronary venous bypass. I. Analysis of variance studies]. AB - In 36 patients, 3 months after aortocoronary venous bypass (ACVB) surgery to the anterior interventricular branch of the left coronary artery, the density-time curve was assessed in the aorta and the ramus interventricularis anterior ACVB by computed tomography (CT). Mean values for area of time-density curve (1539 HU x sec.), effective width (10.9 sec.), peak CM concentration (140 HU), maximum increase of density (71 HU/sec.) and maximum decrease (53 HU/sec.) showed a significant (p greater than 0.01) difference between the results in aorta and the anterior interventricular branch ACVB for all parameters, excepting the effective width. We conclude that assessment of time density-curve parameters in the ramus interventricularis anterior ACVB is feasible; however, the results are significantly different from those obtained in the aorta. Clinical relevance of these data will be established in a second part of the study. PMID- 2548242 TI - [Fine structure analysis of the petrous bone using high-resolution thin-layer computed tomography. 2: Place and success of CT in comparison with conventional x ray diagnosis of petrous bone fractures]. AB - Thirty patients with suspected petrous fractures following trauma were examined by high resolution CT and by conventional radiography. CT proved the most sensitive method for demonstrating petrous fractures and their complications. CT increased the rate of diagnosis by 17%. All fractures seen on CT could be classified, but accurate classification on the basis of conventional radiographs was only possible in 52% of cases. High resolution CT is superior in demonstrating damage to the labyrinth and the ossicles. Indications for conventional tomography are confined to atypical fractures with a course unsuitable for CT. PMID- 2548240 TI - [Diagnosis of postpartum complications using CT and MRT]. AB - Severe puerperal infections, iliacal and ovarian vein thrombosis are post-partum complications which are potentially life-threatening. They require prompt medical treatment. CT and MR enable diagnosis by noninvasive methods. PMID- 2548243 TI - [Observations on the Camurati-Engelmann syndrome. Demonstration of changes of the petrous bone using high-resolution computed tomography]. AB - In Camurati-Engelmann disease HR-CT can impressively demonstrate the involvement of the skull base and also correlate well with the clinical symptoms of deafness, vestibular disturbances and facial paralysis. Since conservative treatment with corticosteroids, calcitonin and diphosphonates is hardly successful and compression of the cranial nerves practically absent, surgical treatment with decompression should be considered. Differential diagnosis against other bone affections with deafness depends on age, typical x-ray findings, local distribution and clinical signs. PMID- 2548245 TI - [Spinal changes in neurofibromatosis in children]. AB - A systematic radiologic study of the skeletal system of 72 children with different forms of neurofibromatosis (NF) revealed a high proportion of spinal defects. Scoliosis was found in 46%, anomalies of the vertebrae in an even higher percentage, particularly cleft spine in 25%, wedge-shaped vertebrae in 22% and scalloping of vertebrae in 40%. Radiographic examination of children with even minimal signs of NF was helpful in confirming the diagnosis, staging of patients and obtaining additional hints in regard to the long term prognosis of children with neurofibromatosis. PMID- 2548244 TI - [MR tomography following lumbar disk surgery: differential diagnostic potentials using Gd-DTPA]. AB - 55 patients were examined by magnetic resonance imaging after lumbar disk operation. The results indicate that with Gd-DTPA there is a significantly increased signal intensity along the surgical approach directly after surgery and during the early postoperative period. Whereas herniated or sequestered disk did not enhance with Gd-DTPA, in epidural fibrosis - regardless of the age of the scarification - diagnostic enhancement was obvious. In 82 percent nerve roots engulfed by scar tissue were differentiated after Gd-DTPA; even enlarged nerve roots and changes of arachnoiditis could be differentiated from each other. PMID- 2548246 TI - [Pathologic changes in the hindfoot angle following fracture of the calcaneus]. AB - Forty fractured calcanei (13 without and 27 with involvement of the joint) were examined by CT and the angle of the hind foot measured. Following a typical compression fracture there is a negative calcaneus-valgus angle, i.e. varus deformity of the tuberosity fragment. The sustentaculum angle is increased. There are changes in the talo-calcaneal and calcaneo-cuboidal angles. The length ratio of calcaneus to talus is reduced. Reproducible measurements make it possible to compare the results of operative correction and of changes in shape following conservative treatment, e.g. during weight-bearing. PMID- 2548247 TI - [Synchronous pulse changes in contrast density during carotid angiography]. AB - The relationship between maximal contrast density in DSA and blood flow in doppler sonography was examined in tube model studies and in carotid angiography. During pulsatile flow, pulse-synchronous changes of contrast density can be registered. The maxima of opacification coincide with the doppler-sonographic phase of re-increasing blood flow after a preceding depression. This is caused by the accumulation of contrast medium at the tip of the catheter during diastole and the subsequent dilution of contrast medium by the increasing blood flow during systole. In model studies as well as in angiography, the time interval between the doppler-sonographic maximum of flow and the maximum of contrast density is very constant and is prolonged with increasing distance from the tip of the catheter. This latency between 2 maxima which are registered at the same region of interest, is caused by the higher speed of propagation of the flow wave compared to the actual movement of the blood corpuscles and contrast medium. The understanding of the relationship between cardiac action, blood flow, and contrast density may improve the functional interpretation of angiograms. PMID- 2548248 TI - [Psychomotor epilepsy--a comparison of CT and MR in 100 patients]. AB - The value of CT and MR was studied in 100 patients with temporal lobe epilepsy. Axial CT scans were obtained before and after contrast injection. Coronary MR scans were carried out with T1-(SE 400/30, GE 315/14) and T2-weighted sequences (SE 1600/30 + 70). A circumscribed lesion was demonstrated in fifteen patients by CT and in 25 patients by MR. With the exception of a small area of calcification, all lesions seen on CT could also be recognized on MR. In four patients CT, MR and pathologic specimen could be compared following partial resection of temporal lobe. Localized glial reactions, which were not seen on CT, produced a signal difference on MR. Better sensitivity and improved demonstration of the temporal lobes makes MR the method of choice in the diagnosis of temporal lobe epilepsy. PMID- 2548249 TI - [Space-occupying lesions of the pineal region in magnetic resonance tomography]. AB - Preoperative CT and MRI studies of 24 patients with space-occupying lesions of the pineal region were analysed. MRI demonstrated all 24 lesions, whereas CT could detect only 20 out of 24. MRI was superior to CT in assessment of size, localisation and relation to adjacent structures. No significant difference was found between CT and MRI in preoperative prediction of tumour histology. PMID- 2548250 TI - [Contrast medium enhanced MRT of tumors of the posterior fossa in children and adolescents]. AB - The results of contrast-enhanced MR in 24 children and adolescents with tumours of the posterior cranial fossa are presented. During initial diagnosis, the majority of tumours showed intensive enhancement with improved demarcation from neighbouring structures. The solid component of a cystic tumour, even when small and not visible on the original scan, could be demonstrated in all cases by means of gadolinium-DTPA. Post-operatively, T2-weighted scans regularly showed areas of high signal strength at the margin of the resection; in the absence of a mass or demonstrable progression, the significance of this is uncertain. In these cases gadolinium-DTPA, because of the function of the blood-brain barrier, greatly increases accuracy in demonstrating residual or recurrent tumours. PMID- 2548251 TI - [Cerebral MRT in neurofibromatosis: gliosis versus neoplasia?]. AB - 15 patients aged 1-39 years with documented neurofibromatosis had MR examinations of the cerebrum within the scope of a basic diagnosis and therapy programme. Clinical examination did not lead to pathologic findings for 7 of the patients; 3 patients suffered from general developmental disabilities. A neurologico psychiatric examination showed pathological findings in 5 patients. Signal intense foci in proton density and T2-weighted MR images were found in the globus pallidus, thalamus, hippocampus, cerebellum and midbrain. In 2 patients, these foci could be found as well in T1-weighted images. Differentiation between gliosis areas and low grade astrocytomas was not possible in MR. PMID- 2548252 TI - [Osteoid osteoma: an unfavorable localization causes difficulties in diagnosis]. PMID- 2548253 TI - [Fibroleiomyoma of the end phalanx of the left ring finger]. PMID- 2548254 TI - Saccular aneurysm of the aortic arch presenting as a nonenhancing mass on CT scanning. Case report. PMID- 2548255 TI - [Computed tomography of a neurinoma of the gastric wall]. PMID- 2548256 TI - Tumoral calcification in association with renal osteodystrophy. PMID- 2548257 TI - [Manifestation of a highly malignant non-Hodgkin's lymphoma of a spinal ganglion]. PMID- 2548258 TI - [A rare manifestation of a primary malignant fibrous histiocytoma of the breast]. PMID- 2548259 TI - [The left ovarian vein as a functional renal vein]. PMID- 2548260 TI - Bilateral Paget-von Schroetter syndrome including superior caval vein. PMID- 2548261 TI - [Sensitive painful polyneuropathy probably caused by poisoning by perchloroethylene. Apropos of 1 case]. PMID- 2548262 TI - [Malignant retroperitoneal fibrohistiocytoma]. PMID- 2548264 TI - [Role of combined etiology (alcohol and B virus) in the development of hepatocarcinoma in subjects with liver cirrhosis]. AB - 107 (8.2%) of 1297 patients with liver cirrhosis admitted to our Clinic between 1980 and 1987 had hepatocellular carcinoma (HCC). This disorder was more frequent in patients with liver cirrhosis of dual etiology, alcoholic and post-hepatitic (9.9%), than in those with liver cirrhosis of unknown (7.82%), alcoholic (9.35%) or post-hepatitic (5.16%) etiology. The mean age of the patients with HCC of dual etiology (51.2 years) was significantly lower than that of the other three groups (61.0, 62.0 and 64.5 years, respectively; p less than 0.001). The conclusion is that in patients with liver cirrhosis of dual etiology the onset of HCC is frequent and precocious. This fact must be considered in epidemiological studies, in the prognosis and in the clinical management of the patient since early diagnosis is of importance in deciding therapy. PMID- 2548263 TI - [Transitory efficacy of potassium perchlorate in hypothyroidism caused by amiodarone]. AB - We report a case of a patient with hypothyroidism due to amiodarone. The short term administration (1 g/die for 10 days) of potassium perchlorate (KClO4) led to normalization of serum thyroid hormone concentrations and marked reduction of thyrotropic hormone. The reduction of KClO4 (400 mg/die) and its following withdrawal led to reappearance of hypothyroidism signs. No side-effects or toxic reactions occurred during KClO4 therapy. This anion competitively inhibits thyroid iodide transport, reducing intrathyroidal iodide content and removing thyroid hormone synthesis inhibition. We suggest KClO4 therapy when amiodarone associated hypothyroidism impairs a pre-existent cardiac disease and when a quick restoration of euthyroidism is necessary. Nevertheless, we emphasize that its effect can be transitory in cases of short-term treatment or low doses. PMID- 2548265 TI - Ca2+ antagonists and db-cAMP sustain a rise in renal blood flow induced by acetylcholine in indomethacin-treated dogs. AB - Renal arterial infusion of acetylcholine (ACh) in the dog normally produces a sustained rise in sodium excretion (UNaV) and in renal plasma flow (RPF). When prostaglandin (PG) synthesis is inhibited, ACh induces only a transient increase in UNaV and RPF followed by a progressive decline in UNaV and RPF, and a rise in renin secretory rate (RSR). Renal arterial infusion of PGE2 but not a vasodilator such as bradykinin restored the response to ACh to normal in indomethacin (Indo) treated dogs. During renal arterial infusion of dibutyryl cyclic AMP (6 mg/min), ACh also produced a sustained increase in UNaV and RPF despite an inhibition of PG synthesis by Indo. Renal arterial infusion of verapamil (60 micrograms/min) or diltiazem (60 micrograms/min) also prevented the subsequent fall in RPF when ACh was infused; RSR, however, did not show a rise. The results suggest that synthesis of PGE2 with stimulation of cAMP is required for sustained ACh action. When PGE synthesis is inhibited, ACh may produce renal vasoconstriction by increasing intracellular Ca2+ concentration. The partial effect of calcium channel blockers suggests that release of calcium from intracellular stores as well as calcium entry may mediate the response. PMID- 2548267 TI - The effectiveness of repeated cardiopulmonary resuscitation training in a school population. AB - At the end of a study program, evaluating the feasibility and the effectiveness of a unique training session on a school population, the majority of the students were asking for additional training opportunities. We therefore set up the present study with the purpose of evaluating skills, knowledge and attitude concerning CPR, after respectively one and two training sessions. 265 students from 4 different school levels were trained. 6 months later 134 answered a questionnaire and were again trained in CPR, 129 students answered the same questionnaire and were tested for their skills in CPR. Ten months later 75 students who had two training sessions answered again the questionnaire and 65 among them were tested for their skills. The two training sessions were identical, given by lay teachers priorly instructed in CPR, and consisted of a video-program and practical demonstration, followed by individual practice on training manikins. Both training sessions lasted 100 min. Evaluation of skills was performed by emergency physicians not involved in the training. Seventeen different items, representing each step in CPR were scored. Repeated training induces significant improvement of total skill scoring, without significant difference between boys and girls, but with improvement of scoring with class level. When looking at the different steps, the improvement in scoring is most impressive in certain steps which scored poorly after one training session, such as backward tilt of the head, a keystone in CPR. The steps concerning mouth-to mouth breathing and external thoracic compressions reach, 10 months after the second training, an average of 1.6 out of 2 (80% correct) as compared to 1.44 out of 2 (71.9% correct) after one training. Knowledge concerning CPR does not increase significantly after the second training session. The time lapse of 10 months since the second training session may have played a role, although the methodology excluding interactive instruction may also explain this discrepancy. The influence on attitude shows that fear to apply CPR increased significantly after one training session and does not significantly lower after the second training. This attitude seems to be rather person-linked, for no correlation was found with age, theoretical knowledge or practical skill scoring. We have no way of knowing whether the statement concerning fear to apply CPR will correspond with such an attitude when confronted with a concrete emergency situation. PMID- 2548268 TI - Increased hyaluronic acid production in lung--a possible important factor in interstitial and alveolar edema during general anesthesia and in adult respiratory distress syndrome. AB - The tremendous swelling potential of hyaluronic acid (HA) is well established, however, the possible role of HA for interstitial and alveolar edema of the lung during general anesthesia and in adult respiratory distress syndrome (ARDS) has not received attention. Bronchoalveolar lavage (BAL) was therefore performed on 28 apparently lung-healthy patients; first 24 h prior to induction of general anesthesia and then--in the same lung segment--approximately 30 min after anesthesia induction with endotracheal intubation and mechanical ventilation with N2O in O2. The second BAL was undertaken prior to surgery. Another group of 11 patients with ARDS was also subjected to BAL. A third control group of 10 patients was investigated with BAL 24 h apart to see if a prior BAL influenced the second BAL. Prominent findings were that induction of general anesthesia was associated with significantly (P less than 0.001) increased amount of HA in BAL, on average 77 micrograms/l (range 24-181 micrograms/l) as compared with HA-level 24 h before anesthesia, where all values were below 20 micrograms/l. ARDS patients ranged from 72 to 1636 micrograms/l with a mean of 515 micrograms/l, which was significantly (P less than 0.001) higher than HA-level after induction of anesthesia. The control group that had BAL of the same lung segment 24 h apart did not show any increase in HA-level during the second BAL; with HA-level below 20 micrograms/l.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548266 TI - Comparative regional distribution of angiotensin-I-converting enzyme in the rat, rabbit, dog, monkey and human kidneys. AB - Kidney is the main source of the production of renin and angiotensin, while also being one of their main target organs. This study was designed to determine the regional distribution of angiotensin-I-converting enzyme (ACE) in the kidney using a biochemical approach. Interspecies variations were analyzed in human, monkey, rabbit, dog and rat kidneys. Kidney ACE content differed among species with decreasing contents as follows: rabbit greater than human greater than monkey greater than dog greater than rat. In rabbit, human, monkey and dog kidneys, we observed predominant cortical distribution of ACE compared with the medulla or papilla; median cortex/papilla ACE activity ratio was 19, 14, 9 and 7 for the rabbit, human, dog and monkey, respectively. In rat kidney, ACE predominantly distributes in the outer medulla, while cortex ACE content appears to be low. The difference in ACE distribution in the rat kidney and to a lesser extent in the dog kidney when compared to rabbit, monkey or man should be taken into account when extrapolating to the human renal hemodynamic studies, which are frequently performed in rats or dogs. PMID- 2548269 TI - Complications associated with barbiturate therapy. AB - Fifty-six patients with elevated intracranial pressure caused by cerebrovascular accident, head injury, etc., were the subjects of this study. They were divided into three groups: low dose barbiturate therapy (15 patients), high dose barbiturate therapy (24 patients), and control group (17 patients). Barbiturate therapy was instituted using thiamylal, and the complications caused by barbiturate therapy were recorded. In the control group, complications occurred in the liver of two patients, but there were no renal or pulmonary complications. Pulmonary, renal, and hepatic complications were common in the barbiturate groups. Complications in the high dose therapy group were significantly more common than in the control group. Opportunistic infections occurred in ten patients, with seven patients having pneumonia. Only one patient, with pneumonia, was seen in the control group. The deaths of three patients were influenced by complications associated with barbiturate therapy, while the single death in the control group was not associated with the complication of barbiturate therapy. PMID- 2548270 TI - Prehospital cardiac arrest--a critical analysis of factors affecting survival. AB - During a 10-year period, 5631 cardiac arrests were treated in our paramedic system. In all, 4216 resuscitations were attempted, of which 533 (12.6%) resulted in saves, defined as hospital discharges. Patients presenting with an initial rhythm of coarse ventricular fibrillation or ventricular tachycardia were found to have significantly increased save rates in comparison to those presenting with an initial arrest rhythm of asystole/fine ventricular fibrillation or electromechanical dissociation (P less than or equal to 0.01). When controlling for witnessed arrest, 303 of 1905 (15.9%) of all witnessed arrests were saves vs. 230 of 2311 (10%) of unwitnessed arrests (P less than or equal to 0.01). Witnessed bystander/first responder external cardiac compression- cardiopulmonary resuscitation (ECC-CPR) was found not to influence save rate. One hundred eighty one of 1248 bystander/first responder witnessed arrests (14.5%) who had external ECC-CPR initiated before paramedic advanced life support arrival were saves, compared with 38 of 252 (15.1%) who had no ECC-CPR initiated until paramedic arrival; this was not statistically significant. Advanced life support response times in saved patients with witnessed cardiac arrests were analyzed. Ninety-five percent of all saves had a response time of less than 10 min. We conclude that, when evaluating the effectiveness of CPR, the variables of witnessing of arrest, presenting arrest rhythm, and respective response times must be controlled or analyzed. PMID- 2548271 TI - Endobronchial administration of emergency drugs. AB - Endobronchial emergency drug therapy is reviewed. Although intravenous drug administration is always preferable endobronchial application is a comparatively safe alternative when venous access is not available because an endotracheal tube is usually inserted during CPR. The optimal procedure remains to be clarified, however. Drugs and dosages currently being recommended for endobronchial use are described. With the reservations mentioned above the following recommendations can be made for the endobronchial administration of drugs to adult emergency patients: (1) Endobronchial administration of drugs is indicated in cases of cardiac arrest when an initial examination after the start of mechanical measures suggests that intravenous application will be difficult. This is also the case for emergency patients without cardiac arrest who are already intubated, and for whom the immediate administration of emergency drugs, such as lidocaine or atropine, is urgent and for whom venous injection cannot be carried out or cannot be carried out sufficiently quickly. We cannot at present share the view that the primary endobronchial administration of epinephrine is to be regarded as the treatment of first choice in every case of asystolic cardiac arrest (J. Schuttler et al., Anasth. Intensiwther. Notfallmed., 22 (1987) 63), as in most cases peripheral venous access can readily be established, thus avoiding the possible disadvantages of endobronchial administration (prolonged tachycardia, hypertension, arrhythmia from the depot effect, and also additional impairment of the pulmonary gas exchange). However, the washing-in of the administered drug is important after peripheral venous administration. (2) For adults, the present recommendation for the i.v. administration of epinephrine during resuscitation is 0.5-1 mg (7.5-15 micrograms/kg). Animal experiments show that under stable circulatory conditions ten times as much epinephrine must be administered endobronchially as intravenously in order to produce comparable pharmacodynamic effects or plasma levels. During resuscitation the corresponding value is approximately five times. Nevertheless, because of the dangers from the depot effect, an endobronchial dosage of 2-3 mg should not at present be exceeded. The best method probably consists of a combination of an initial restricted endobronchial dose with, if necessary, a further dose through a venous site which has been established in the interim. This recommendation seems to be reasonable because, as described, there are no exact data on the optimal dosage for epinephrine, even for i.v. administration (3) In the presence of a stable circulation the endobronchial dosag PMID- 2548273 TI - A continuous hemorrhage model of fatal hemorrhagic shock in swine. AB - We studied the effect of bleed rate on survival time and hemodynamics in a continuous hemorrhage model of fatal hemorrhagic shock in lightly anesthetized swine. Fasted immature swine (12-16 kg) were sedated with intramuscular (i.m.) ketamine, endotracheally intubated, anesthetized with halothane (0.75%), nitrous oxide, and oxygen, and then prepared for experimentation by placement of a pulmonary artery thermodilution catheter, femoral arterial and venous catheters, and by splenectomy. After instrumentation, halothane was discontinued and sedation was maintained with nitrous oxide and intravenous lorazepam. Thirty minutes later, the animals were bled continuously at 1.0 ml/kg per min (n = 8, Group I) or 1.25 ml/kg per min (n = 8, Group II) by a roller pump connected to the femoral arterial catheter. Hemodynamic parameters were recorded every 15 min until death occurred. Mean survival time was 50.2 +/- 3.0 min in Group I and 39.8 +/- 3.2 min in Group II (P less than 0.001). There was a stepwise decrease in blood pressure and cardiac index consistent with progressive hemorrhagic shock. This model results in reproducible survival times with small standard deviations. Although the animals are lightly anesthetized and the experiments are performed acutely, the hemodynamic responses and survival times observed are similar to those reported in previous studies of chronically instrumented, unanesthetized swine. This model may be more practical than unanesthetized, chronically instrumented swine models for evaluating the effects of various interventions on survival time and hemodynamics in acute hemorrhagic shock. PMID- 2548272 TI - Continued circulatory support: effect of epinephrine or dopamine on 24-hour survival and neurologic function in dogs. AB - The effects on 24-h survival and neurologic function were compared following continued postresuscitation circulatory support with epinephrine or dopamine. Cardiopulmonary arrest was induced by ventricular fibrillation. After 10 min, resuscitation efforts were initiated including i.v. infusion of either epinephrine (6 micrograms/kg per min, 11 dogs) or dopamine (10 micrograms/kg per min, 14 dogs) for continued circulatory support. There was no difference detected in duration of circulatory support, although dogs receiving epinephrine required more lidocaine (3.3 +/- 0.4 vs. 1.8 +/- 0.3 mg/kg, P = 0.005). Likewise, there was no statistically significant difference detected in MAP or HR between groups at any time tested. However, dogs receiving epinephrine had significantly worse neurologic function at 6 and 12 h postarrest. Mean survival time (20.3 +/- 1.2 vs. 15.3 +/- 1.9 h, P = 0.028) and overall survival (P = 0.027, survival curve analysis) were significantly longer for dogs receiving dopamine. Plasma glucose in the first 6 h postarrest was significantly higher in dogs receiving epinephrine (P = 0.006). These results suggest that the use of epinephrine for continued vasopressor support in cardiopulmonary resuscitation may contribute to decreased survival and poorer neurologic function in this controlled experimental setting. It is reasonable to propose that similar responses to these commonly used circulatory support agents occur clinically. Therefore, continued vasopressor support with dopamine rather than epinephrine may be justified in the setting of cardiac resuscitation. PMID- 2548274 TI - Long-term follow-up of in situ carcinoma of the breast with special emphasis on clinging carcinoma. AB - Forty-two cases of in situ duct (28 cases) and lobular (14 cases) carcinoma were identified after a review of 4,397 "benign" breast biopsies obtained from the files of departments of pathology of a small area of Northern Italy, between 1965 and 1971. None of the patients with in situ lobular carcinoma developed an invasive carcinoma. On the other hand, three patients with in situ duct carcinoma (one comedo and two clinging type) developed subsequent invasive carcinoma. Therefore, it appears that clinging carcinoma has the same biological behavior as other types of small in situ ductal carcinoma; the likelihood of our patients developing invasive breast carcinoma is four times greater than that of the general population. PMID- 2548275 TI - [Diffuse interstitial pneumopathy caused by herpes simplex virus type 1 in a 77 year-old patient]. PMID- 2548276 TI - Prospects for development of a rotavirus vaccine against rotavirus diarrhea in infants and young children. AB - Major advances have been made in elucidating the etiologic agents of severe infantile diarrhea, and it is clear that rotaviruses are the single most important etiologic agents. Progress in the development of rotavirus vaccine candidates has also moved swiftly with the "Jennerian" approach, in which a related live, attenuated rotavirus strain from a nonhuman host is used as the immunizing antigen. If this strategy is not effective against all rotavirus serotypes, reassortant rotaviruses hold great promise for the development of a multivalent vaccine. Field trials with the "Jennerian" approach vaccines are under way, and phase 1 trials with the reassortants have been initiated. PMID- 2548277 TI - Evidence of central nervous system involvement in chronic demyelinating neuropathies associated with "benign" gammopathies. AB - Recent papers underline the possible involvement of the central nervous system when an acquired peripheral demyelinating disease occurs and vice-versa. We describe five patients with chronic polyneuropathy and "benign" gammopathy, monoclonal (IgM-K, IgA-k, IgG-k) in three cases and polyclonal (IgG, IgM) in two cases; the monoclonal gammopathies were detected in cases of peripheral nerve disease. Three patients showed tremor and signs of pyramidal system impairment when the peripheral damage had improved or was stable. All cases underwent a longitudinal assessment according to clinical, CSF, EMG-ENG, neuroradiological and pathological criteria. The MRI finding always showed multiple alterations of encephalic white matter. When related to neurophysiological and pathological data supporting a chronic demyelinating neuropathy, such results point to possible encephalic involvement in chronic polyneuropathies due to a pathogenetic mechanism common to both. PMID- 2548278 TI - Inhibition of human natural killer cell and lymphokine-activated killer cell cytotoxicity and differentiation by vitamin D3. AB - Recent data suggest that vitamin D3 may be capable of immunoregulation after it is converted to an active metabolite, 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). The effect of vitamin D3 and 1,25(OH)2D3 on human natural killer (NK) cells and their activation by interferon (IFN) and interleukin 2 (IL-2) was investigated. Vitamin D3 and 1,25(OH)2D3 inhibited NK cytotoxicity in a dose-dependent manner. Pretreatment of non-adherent (NA) cells at 37 degrees C for 18 h with the vitamins also led to inhibition of NK activity. Both the inhibition of NK lysis and pretreatment of NA cells were dependent on the concentrations of fetal calf serum (FCS) in the medium. The inhibition of NK activity was less effective in the presence of 10% FCS than with 1% FCS. Vitamin D3 inhibited both IFN and IL-2 activation of NK activity. However, increasing doses of IL-2 were able to abrogate the inhibition caused by vitamin D3. Vitamin D3 was able to inhibit NK activity of phytohaemagglutinin and IL-2-activated cells, and also inhibit the proliferation and lymphokine-activated killer activity induced by IL-2. NA cells pretreated with vitamin D3 did not respond well to IL-2. NA cells pretreated with low doses of IL-2 were sensitive to inhibition by vitamin D3 while those pretreated with high doses of IL-2 were not. The data presented suggest that vitamin D3 and 1,25(OH)2D3 inhibit NK activity and LAK cellular differentiation. PMID- 2548279 TI - Conserved folding in retroviral proteases: crystal structure of a synthetic HIV-1 protease. AB - The rational design of drugs that can inhibit the action of viral proteases depends on obtaining accurate structures of these enzymes. The crystal structure of chemically synthesized HIV-1 protease has been determined at 2.8 angstrom resolution (R factor of 0.184) with the use of a model based on the Rous sarcoma virus protease structure. In this enzymatically active protein, the cysteines were replaced by alpha-amino-n-butyric acid, a nongenetically coded amino acid. This structure, in which all 99 amino acids were located, differs in several important details from that reported previously by others. The interface between the identical subunits forming the active protease dimer is composed of four well ordered beta strands from both the amino and carboxyl termini and residues 86 to 94 have a helical conformation. The observed arrangement of the dimer interface suggests possible designs for dimerization inhibitors. PMID- 2548280 TI - The expanding role of cisplatin in the treatment of non-small-cell lung cancer. AB - Cisplatin has been employed in the treatment of non-small-cell lung cancer (NSCLC) since the late 1970s. Current evidence suggests that it may be the most effective single drug for the treatment of NSCLC patients. The overall response rate as a single agent is 21%, though responses are higher in previously untreated patients given high doses. Cisplatin combinations are the most effective therapy for advanced NSCLC. Two-drug combinations with a vinca alkaloid or etoposide consistently provide the best overall survival with acceptable toxicity. These two-drug combinations improve survival by 7 to 17 weeks compared with no chemotherapy. Cisplatin-based combinations improved survival by a greater degree in locally advanced patients (Stages IIIA and IIIB) when given with chest radiotherapy. There is evidence that cisplatin-based combinations also prolong survival when given after surgery for resected stage II and IIIA patients. In summary, cisplatin-based chemotherapy regimens are the preferred systemic therapy for NSCLC patients and prolong survival to a small degree. PMID- 2548281 TI - Disseminated testicular cancer: current chemotherapy strategies. AB - Disseminated testis cancer is a landmark adult solid tumor because of its high curability. In the last decade and a half, a number of clinical trials have been completed or are ongoing. In addition, long-term follow-up of some earlier trials is now available. This report summarizes the current treatment approaches, paying particular attention to the role of cisplatin. PMID- 2548282 TI - Cisplatin for small-cell lung cancer. AB - Small-cell lung cancer (SCLC) is a tumor highly sensitive to chemotherapy for which combination chemotherapy is the cornerstone of treatment. Antineoplastic activity has been shown for several agents, usually in the setting of minimal prior therapy. Active agents have minimal activity when used after previous chemotherapies. Cisplatin, which has only modest activity for SCLC, has not been adequately tested in the setting of minimal prior therapy, but is probably very active. Furthermore, response to cisplatin is probably dose-dependent. Synergy between agents is very important in treatment strategies, and cisplatin is synergistic with multiple agents. The EP combination (cisplatin, etoposide) is particularly synergistic for SCLC. Unlike most other combinations, EP produces consistent responses as a salvage regimen. When used as an initial treatment regimen or to alternate with other combinations, EP approximately produces the "state-of-the-art" anticipated results. When EP is administered with concurrent chest irradiation in limited disease, it produces response and survival results similar to more aggressive regimens. Thus, EP plus chest irradiation is a reasonable combination for patients not entering investigational studies, and EP may be the foundation for more aggressive combinations. CEP (high-dose EP, cyclophosphamide) caused an increase in response frequency in extensive disease. We added cisplatin to the combination of cyclophosphamide, doxorubicin, and etoposide (PACE). This four-drug combination pilot study was stopped early because of toxicities, and follow-up now suggests that survival may be prolonged. Further study of this and similar aggressive combinations appears warranted, and the use of colony-stimulating factors may allow for acceptable toxicity and further dose escalation. PMID- 2548283 TI - [Lung cancer. The language of stages]. PMID- 2548284 TI - [Lung cancer. Chemotherapy]. PMID- 2548285 TI - [The content of tyrosine in rat luteal cell and the changes caused by hCG, cAMP and progesterone]. AB - The ovaries taken from the immature female rats primed with PMSG-hCG were digested with collagenase-DNAase solution to obtain the luteal cell suspensions. Luteal cells were then incubated with different doses of hCG, cAMP or progesterone for 1.5 hours. The contents of tyrosine in luteal cell suspensions were determined by thin layer chromatogram using dual-wavelength chromatogram scanner. It was found that the content of tyrosine in rat luteal cell suspensions was 1.41 + 0.24 nmol/L/10(6) cells after one hour incubation. hCG, cAMP and progesterone all could significantly increase the content of tyrosine in the suspensions, suggesting the release of "endogenous tyrosine". The increase of tyrosine was not due to increased transformation of phenylalanine, the precursor of tyrosine, and increased protein synthesis, because both phenylalanine and cycloheximide failed to influence such increase. PMID- 2548286 TI - [Ionic mechanism of noradrenaline-induced membrane potential changes of neurones in toad dorsal root ganglion]. AB - The membrane conductance and reversal potential were determined for neurones in toad dorsal root ganglion (DRG) with intracellular recording technique during depolarization or hyperpolarization induced by noradrenaline (NA). The effects of blocking agents for potassium or calcium channels on NA-induced membrane potential responses were examined. In 15 neurones, the NA-induced depolarization was accompanied by a 32.6% decrease of membrane conductance; in other 4 neurones, the depolarization was accompanied by an initial increase and subsequent decrease in membrane conductance. The NA hyperpolarization was associated with an increase of membrane conductance by 16.2% (n = 8). The mean reversal potential of NA induced depolarization was -88.5 +/- 0.9 mV (means +/- SE, n = 4). The NA-induced hyperpolarization was nullified at -89 to -92 mV of membrane potentials (n = 3). Tetraethylammonium superfusion enhanced NA depolarization amplitude by 73.7 +/- 11.9% (means +/- SE, n = 7) and depressed NA hyperpolarization amplitude by 40.5% (n = 4). Intracellular injection of CsCl increased phenylephrine-induced depolarization by 34.5% (n = 4). MnCl2 superfusion decreased the amplitudes of NA induced depolarization by 50.5 +/- 9.9% (means +/- SE, n = 10), and of NA-induced hyperpolarization by 89.5 +/- 4.9% (means +/- SE, n = 7) respectively. The results suggest that the depolarization or hyperpolarization induced by NA might be mediated by the alteration in activation of K+ or Ca2+ channels. PMID- 2548287 TI - [Experimental studies of healing process on compound blocks of hydroxyapatite (HAP) particles and tricalcium phosphate (TCP) powder implantation in rabbit mandible--comparison of HAP/TCP ratios and plastic methods]. AB - Bone substitutes consisting in 2 mold-poured compound blocks of HAP:TCP (mixing ratio 1:1) and HAP:TCP (mixing ratio 3:1) and one compound block (mixing ratio 1:1) prepared in a rubber press were implanted in defective areas in rabbit mandibles. The conditions under which the implants were made were identical in all cases. Histological studies and contact microradiography performed on ossification of the surfaces and surroundings of each block, ossification maturation, and the absorptive process of TCP and the extent to which it was replaced by new bone produced the following results. 1. At one week after surgery, new bone had grown along the surfaces of the blocks. It remained in tight contact with the block surfaces as it matured. These findings make it clear that the blocks are superior in terms of biocompatibility and osteoconduction. 2. In the course of time, in the 2 mold-poured compound blocks, the hydrated parts of alpha-TCP, which was a binder, were absorbed and replaced by new bone. At forty eight weeks after surgery, new bone had grown into the pores of the HAP particles to form a bone-HAP composite. 3. Even as late as forty eight weeks after surgery, some of the hydrated parts of alpha-TCP remained unabsorbed; and mold-poured compound blocks (mixing ratio 1:1) retained their shapes. On the other hand, in the mold-poured compound blocks (mixing ratio 3:1), the hydrated parts of alpha-TCP were almost absorbed and replaced by new bone in forty eight weeks after surgery. Amounts of the binder alpha-TCP present appeared to cause differences in biological reactions. 4. In the case of compound blocks of HAP:TCP (mixing ratio 1:1) formed in a rubber press, the binder was beta-TCP. Because of its high sintering properties and great density, absorption and ossification were gradual; and the blocks retained their complete original forms even in forty eight weeks after surgery. 5. Past reports have claimed that beta-TCP is absorbed in the initial stage. But, as is clear from experiments with compound blocks formed by means of a rubber press, even when the binder is beta-TCP in all cases, absorption speed differs depending on the mold used. 6. These results indicate that compound blocks of HAP and TCP used as artificial substitute bone materials have excellent properties of biocompatibility, osteoconduction, and plasticity. In addition, however, they make it clear that, owing to differences in absorption of the binder, ossification speed, and dynamic properties from block to block, it is essential to use the mold that properly suits conditions prevailing in the defective area. PMID- 2548288 TI - Primary mucinous cystadenocarcinoma of the appendix with pseudomyxoma peritonei manifested as a splenic mass. AB - We have reported a case of pseudomyxoma peritonei manifested as a splenic mass in a 38-year-old woman. Upon reviewing previously reported cases of pseudomyxoma peritonei with visceral involvement or extension above the diaphragm, we conclude that such spread of the disease does not significantly alter the prognosis. Furthermore, our findings support the concept that pseudomyxoma peritonei represents the implantation of malignant cells rather than metaplastic transformation of mesothelial cells. PMID- 2548289 TI - Bibrachial palsy due to paraneoplastic encephalomyelitis. AB - Bibrachial palsy is a distinctly unusual pattern of muscular weakness. We have described a woman who had a slowly progressive clinical course, with profound bibrachial palsy and neurogenic respiratory failure, but neurologically normal lower extremities. Autopsy revealed encephalomyelitis and a small previously undetected oat cell carcinoma with inflammatory infiltration. The pathologic features are consistent with an autoimmune process. PMID- 2548290 TI - Cytomegalovirus and appendiceal perforation in a patient with acquired immunodeficiency syndrome. AB - We have reported the case of a patient with AIDS who had appendiceal perforation and periappendiceal abscess. Extensive infection with cytomegalovirus was found in the appendix; no other pathogen could be implicated in the perforation. This case emphasizes the broad spectrum of disease due to cytomegalovirus, particularly in patients with AIDS and other immunodeficiencies. PMID- 2548291 TI - Biochemical and genetic analysis of toxic effect of HOE 15030 in mammalian cells. AB - HOE 15030 inhibited the growth of BHK cells at concentrations that did not inhibit their nuclear DNA and RNA syntheses. When BHK cells were cultured in the presence of 30 micrograms/ml of HOE 15030, cells were arrested in the G1 phase after one or two cell divisions. After removal of the drug, cells progressed through the G1 to the S phase. HOE 15030 inhibited the activities of both topoisomerases I and II in vitro. To determine the target molecule of HOE 15030 in cells, we isolated a HOE 15030-resistant (HOEr) mutant of BHK cells. The HOEr cells exhibited cross-resistance to ethidium bromide, acriflavine, and rhodamine 123, and slight cross-resistance to 4'-dimethylepipodophyllotoxin-4-(4,6-O ethylidine-beta-D-glu copyranoside) (VP-16) and adriamycin, but not to chloramphenicol, oligomycin, novobiocin, colchicine, or vinblastine. The uptake and retention of rhodamine 123 by HOEr cells were lower than those by BHK cells. Mitochondrial DNA synthesis of HOEr cells was more resistant to HOE 15030 and ethidium bromide than that of wild-type cells. These results indicate that the resistance of HOEr cells to drugs is due to reduced uptake or accumulation of the drugs by mitochondria and suggest that the mitochondria are the main target of HOE 15030 in cells. PMID- 2548293 TI - Insulin-like growth factor I receptor gene is concordant with c-Fes protooncogene and mouse chromosome 7 in somatic cell hybrids. AB - The insulin-like growth factor I (IGF-1) mediates the actions of pituitary growth hormone in a variety of tissues. Its receptor (IGF1R) displays considerable structural similarity to the insulin receptor. In humans, the IGF1R gene has been mapped near FES, the cellular counterpart of the feline sarcoma virus transforming gene v-fes, at the q25-q26 region of human chromosome 15 (HSA15). Here, we report the mapping of mouse Igf1r to mouse chromosome 7 (MMU7) by somatic cell hybrid analysis. This result, along with the prior assignment of the loci for mitochondrial isocitrate dehydrogenase and FES to human chromosome 15 and mouse chromosome 7, suggest a conserved autosomal synteny group on the distal long arm of HSA15 and in the center of MMU7. PMID- 2548292 TI - Cell genetic evidence of correlation of intracellular translocation of protein kinase C (PKC) and PKC-mediated phosphorylation of 80-kDa protein with mitogenic action of tumor promoters. AB - Recently, we isolated a series of 3T3-L1 cell variants that are unable to respond to mitogenic stimulation by the tumor promoter, 12-O-tetradecanoylphorbol acetate (TPA). Since protein kinase C (PKC) is the major receptor for TPA and appears to play a key role in cellular proliferation, we have examined the distribution of PKC in the parental 3T3-L1 cells and the variant VT-1 cells. PKC was located predominantly in the cytosol of growth-arrested confluent 3T3-L1 cells, and upon TPA treatment it was rapidly translocated into the plasma membrane. In contrast, PKC was located predominantly in the plasma membrane of confluent VT-1 variant cells and was no longer activated by TPA. Two-dimensional gel analysis showed that a Mr 80,000 acidic protein (80-kDa protein) was rapidly phosphorylated in 3T3-L1 cells upon TPA treatment, whereas phosphorylation of this protein was barely detected in VT-1 cells. In growing cultures, the majority of PKC was found in the plasma membrane of both cell lines, and no change occurred upon TPA treatment. Hydroxyapatite column chromatography revealed the presence of alpha type PKC as the major component in both cell lines. These results suggest that the intracellular translocation of alpha-type PKC and the PKC-mediated phosphorylation of the 80-kDa protein may be involved in the mechanism of mitogenic signal transfer. PMID- 2548294 TI - Shaving and dermabrasion of the facial lesions in tuberous sclerosis. A case report. AB - Tuberous sclerosis is a disease characterised by convulsive seizures, mental deficiency and angiofibromas. These angiofibromas are hamartomas consisting of hyperplastic connective and vascular tissue. A case is reported where multiple angiofibromas of the face resulted in significant disfigurement. The lesions were treated by shaving and dermabrasion; the short-term result was very satisfactory. PMID- 2548295 TI - The effect of estrogen on low-density lipoprotein binding kinetics in aortic endothelial cells. AB - The inhibition of atherosclerosis by estrogen has been shown clinically and experimentally, but the mechanism by which this occurs is unknown. Previous studies have shown that estrogen enhances the uptake of low-density lipoprotein (LDL) by bovine aortic endothelial cells (BAEC) while not altering membrane binding at saturating levels of LDL. In this study the effect of estrogen on LDL binding kinetics has been investigated. Computer-assisted Scatchard analysis of binding data suggests a single-site binding model. Estrogen-treated BAEC showed a lower binding affinity (Ka = 2.47 +/- 0.74 E7 M-1) than control cells (1.95 +/- 0.21 E7 M-1) (p = 0.0012). Estrogen-treated cells, however, had a greater binding capacity (Bmax = 1.26 +/- 0.07 E-10M) than control cells (Bmax = 8.49 +/- 0.44 E 11M) (p = 0.0004). The latter was due primarily to a difference in LDL binding at higher concentrations of LDL (greater than 40 micrograms/ml). These findings are consistent with an estrogen-stimulated increase in low-affinity binding of LDL to BAEC, which may not be directly receptor mediated and which appears to enhance the uptake of LDL at higher lipoprotein concentrations. Such alterations in LDL uptake by endothelial cells could influence the formation of atherosclerotic plaque. PMID- 2548296 TI - Postreceptor mechanisms of small-bowel water and electrolyte transport. AB - Neurohumoral agents modulate intestinal transport by interactions with cell membrane receptors. Intracellular second messenger systems implicated in mediation of membrane receptor regulation of cellular events include the phosphoinositide and adenylate cyclase systems. In this study we have investigated the effects of direct postreceptor activation of key components of these systems on intestinal water and electrolyte transport. Rabbit ileal segments (n = 35) were arterially perfused ex vivo with an oxygenated sanguineous solution. The lumen was perfused with an isotonic solution containing 14C polyethylene glycol as a nonabsorbable marker. Net fluxes of H2O, Na+, and Cl- in six experimental groups were calculated for three 20-minute periods: basal, drug infusion, and recovery. The control group had no drug infusion. Two phorbol esters--phorbol 12, 13-diacetate (PDA; 10(-5) mol), and phorbol 12, 13-dibutyrate (PDB; 10(-5) mol)--were used to activate protein kinase C, an important component of the phosphoinositide system. The inactive 4 alpha-phorbol 12, 13-didecanoate (PDD; 10(-5) mol) served as a drug-infused control. Forskolin at two doses (FOR; 10(-5) mol and 10(-6) mol) was used to activate adenylate cyclase. The control and PDD groups had no changes in the flux of water and electrolytes. Both PDA and PDB had proabsorptive effects, with the more lipophilic and potent phorbol ester (PDB) having a more pronounced, significant effect (p less than 0.05). FOR caused significant secretion of H2O, Na+, and Cl- in a dose-dependent fashion (p less than 0.05). These results indicate that direct protein kinase C activation causes a proabsorptive effect and that direct activation of adenylate cyclase causes a secretory effect in the isolated small bowel. The activation status of these second messenger systems has a major influence on the transport state of the intestine. PMID- 2548297 TI - Human cytomegalovirus infection. PMID- 2548298 TI - Hypoxia, almitrine, and peripheral neuropathy. PMID- 2548299 TI - Peripheral nerve function in patients with chronic bronchitis receiving almitrine or placebo. AB - A double blind prospective study of the effect of almitrine bismesylate and placebo on peripheral-nerve function was carried out in 12 patients with chronic bronchitis and arterial hypoxaemia (mean (SD) FEV1% predicted 38 (16), arterial oxygen tension (PaO2) 7.56 (0.76) kPa). Of the seven patients who took placebo, none developed symptoms or signs of peripheral neuropathy. One patient who had abnormal lower limb sensory nerve conduction initially showed improvement of sensory conduction but deterioration in motor conduction during the 12 month study period. Two further patients developed some slowing of motor conduction velocities in their right lateral popliteal nerve. Five patients received almitrine and all showed an improvement in PaO2 (mean from 7.0 to 7.9 kPa). None had symptoms or signs of peripheral neuropathy on entry to the study; one patient had evidence of impaired nerve conduction on electrophysiological testing. Three patients developed symptoms and signs of peripheral neuropathy during the 12 months of the study and a fourth developed peripheral neuropathy at 18 months, having continued to receive almitrine. Studies of nerve physiology showed abnormalities in the lower limbs of all four patients. Recovery was poor, possibly because of the long half life of almitrine. The studies suggest that almitrine may precipitate peripheral neuropathy in patients with chronic obstructive pulmonary disease. Patients should be warned of this potential complication so that the drug can be stopped as soon as symptoms develop. PMID- 2548301 TI - Role of bone marrow in the development of thrombosis. A study of the curative and prophylactic action of heparin and heparin fragment (CY 222). PMID- 2548300 TI - Effect of inhaled leukotriene B4 alone and in combination with prostaglandin D2 on bronchial responsiveness to histamine in normal subjects. AB - The effect of intradermal injection of leukotriene B4 alone and in combination with prostaglandin D2 and E2 and the effect of inhaled leukotriene B4 in combination with prostaglandin D2 were studied in six non-asthmatic men. The intradermal injection of leukotriene B4 (1 microgram) alone caused no immediate or late response in five of the six subjects but greatly potentiated the flare response to intradermal prostaglandin D2 (0.5 microgram) and E2 (0.5 microgram) in all subjects. In contrast, inhaled prostaglandin D2 (6 micrograms) alone and in combination with inhaled leukotriene B4 (12 micrograms) caused no change in the response to inhaled histamine, measured 30 minutes and three and six hours after the inhalation. These findings provide no support for the suggestion that leukotriene B4 has an important role in causing bronchial hyperresponsiveness. The possibility that higher doses of inhaled leukotriene B4 may alter bronchial responsiveness cannot, however, be ruled out. PMID- 2548302 TI - [Hepatitis contagiosa canis (H.c.c.)--2 cases in Austria]. AB - Two cases of H.c.c. which occurred in winter 1987 in Vienna are described. Case one was a female Chow-Chow, 8 weeks of age, that died from the peracute form of the disease. The diagnosis was confirmed by histology and direct immunofluorescence. Case two, a 9-month old female Kuvacz, showed clinical signs of the subacute form of H.c.c. She was hospitalized and therapy was successful. The disease was diagnosed by the typical clinical signs and the raise of antibodies in paired serum samples. Etiology, clinical signs and immunology of H.c.c. are discussed. PMID- 2548303 TI - Toxicity to alveolar macrophages in rats following parenteral injection of nickel chloride. AB - Alveolar macrophages collected by pulmonary lavage from male Fischer-344 rats at intervals (1-72 hr) after NiCl2 injection (62-500 mumol/kg, sc) were tested by several techniques. Within 1 to 4 hr, the macrophages showed morphological and biochemical signs of activation (hypertrophy, ruffled plasma membrane, increased cyclic AMP concentration, and markedly diminished 5'-nucleotidase activity, assayed by concanavalin A inhibition). Functional impairment (reduced phagocytic activity) was first seen at 24 hr; lipid peroxidation (increased malondialdehyde concentration) was not detected until 48 hr. Dose- and time-related effects of NiCl2 on 5'-nucleotidase activity, phagocytic activity, malondialdehyde concentration, and nickel content of alveolar macrophages were observed 24 to 72 hr postinjection. Diminished cell viability occurred only at 72 hr after the highest dosage of NiCl2. In alveolar macrophages from 63NiCl2-treated rats, 63Ni was located primarily in the cytoplasm, based upon liquid scintillation counting and autoradiography; fractionations of macrophage cytosol by gel filtration chromatography showed that 63Ni was bound to several high- and low-molecular weight constituents. This study demonstrates that sc administration of NiCl2 to rats caused nickel uptake into and activation of alveolar macrophages, followed by reduced phagocytic capacity. The alveolar macrophage was a cellular target for nickel toxicity following parenteral exposure to NiCl2. PMID- 2548304 TI - Possible mechanism of chrysotile asbestos-stimulated superoxide anion production in guinea pig alveolar macrophages. AB - Excessive production of active oxygen radicals by macrophages is proposed to play an important role in asbestos-related diseases. The purpose of this study was to examine the capacity and mechanisms of action of various forms of asbestos to stimulate superoxide anion production by guinea pig alveolar macrophages. Chrysotile, but not the amphiboles (crocidolite, anthophyllite, or amosite), stimulated a rapid (less than 1 min) and dose-dependent (2.5-50 micrograms/ml) production of superoxide anion at noncytotoxic doses (2.5 to 25 micrograms/ml). The stimulation of superoxide anion production by chrysotile could be blocked by putative protein kinase C inhibitors (staurosporine, sphingosine, and fluphenazine). Chrysotile also stimulated phosphatidylinositol turnover as measured using 32Pi incorporation into phospholipids, [3H]-diacylglycerol levels, and intracellular calcium mobilization as measured using fura-2 and 45Ca. In addition, pertussis toxin partially blocked chrysotile-stimulated superoxide anion production. We conclude that the mechanism of guinea pig alveolar macrophage stimulation by chrysotile, but not the amphibole asbestos forms, is consistent with a mechanism which is similar to that used by agonists such as N formyl-Nle-Leu-Phe resulting in stimulated phosphatidylinositol turnover, calcium mobilization, and activation of protein kinase C. PMID- 2548305 TI - Determination and metabolism of dithiol chelating agents. VIII. Metal complexes of meso-dimercaptosuccinic acid. AB - Metal complexes of meso-dimercaptosuccinic acid (DMSA) with Pb2+, Cd2+, and Hg2+ were studied by potentiometric and infrared methods. This dimercapto metal binding agent was found to form complexes whose structures are dependent on the metal ion to be complexed. In the cases of Pb2+ and Cd2+, one oxygen and one sulfur act as the donor atoms; in the case of Hg2+, two sulfur atoms act as the donors. The solubilities of all metal chelates were found to be pH dependent. Complexes of cadmium and lead are insoluble in the pH range 1.0 to 7.1, but are solubilized when the noncoordinated sulfhydryl and carboxylic acid groups are ionized. The mercury complex is insoluble in the pH range 1.0 to 3.0. It dissolves when one of the noncoordinated carboxylic acid groups is ionized. The dimethyl ester of meso-DMSA (DiMe-meso-DMSA) was synthesized and its acid dissociation constants were determined (pK1 = 6.38 and pK2 = 8.00). Esterification of the carboxyl groups of meso-DMSA changes its coordination properties in that the two sulfur atoms of DiMe-meso-DMSA are used to coordinate with Hg2+, Cd2+, or Pb2+. Esterification of meso-DMSA also changes its biological properties. DiMe-meso-DMSA, when given to rats 3 days after Cd administration, greatly increased the excretion of Cd via bile. In contrast, meso-DMSA was devoid of such activity. PMID- 2548306 TI - Influence of diethyldithiocarbamate on nickel-induced hepatic metallothionein in rats. AB - The influence of sodium diethyldithiocarbamate (DDC) on 63nickel chloride-induced metallothionein (MT) and alterations in essential metal levels in the liver of rats was investigated. The induction of MT, Zn and Cu levels of the hepatic cytosolic "heat stable fraction" (HSF) by DDC increased with time up to 24 hr. Although MT, Zn and Cu were significantly greater at 17 hr than those at 6.5 hr after 63Ni administration, the Ni level decreased. The treatment with DDC at 6 hr, but not at 10 min, prior to 63Ni administration increased significantly the MT, Zn and Cu contents 17 hr after 63Ni administration over that caused by 63Ni alone at 17 hr., showing a synergistic effect. The induction of hepatic MT by 63Ni or DDC alone was prevented by cycloheximide but not by actinomycin D and seemed to be regulated at the protein synthesis level. PMID- 2548307 TI - NIOSH training in occupational health. PMID- 2548308 TI - CMV infection and vascular rejection in renal transplant patients. PMID- 2548309 TI - Importance of PDGF receptor expression in accelerated atherosclerosis-chronic rejection. PMID- 2548310 TI - Aflatoxins in liver biopsies from Kenya. AB - The results of aflatoxin analysis of 15 needle liver biopsies from a rural hospital in Kenya are reported. Nine of the biopsies were from living subjects and six were post mortem. Blood and urine collected the same day was also analysed for aflatoxin content. Aflatoxins in the liver specimen from living subjects were only found in 3 out of 5 cases of hepatocellular carcinoma, but not in their blood. Aflatoxins were detected in 4 out of 6 post mortem liver specimens. In all three cases of cirrhosis aflatoxins were detected in blood and urine, but only one liver specimen showed aflatoxins. Our findings support the existing theory about the incrimination of aflatoxins in the aetiology of hepatocellular carcinoma, and the possible role aflatoxins may have in the pathogenesis of some forms of liver cirrhosis. PMID- 2548311 TI - Unusual presentation of amoebic liver abscess. AB - Two cases of amoebic liver abscess are described. The unusual, purely left pleuritic, presentation in the first patient is emphasised. Amoebic liver abscess should still be thought of in any patient presenting with an acute febrile illness, upper abdominal and or thoracic manifestations and polymorphonuclear leucocytosis. Such a consideration should obviate unnecessary morbidity, mortality and the injudicious use of ineffective therapeutic agents. PMID- 2548312 TI - Malignant giant gastric ulcer in a Nigerian. AB - A case of malignant giant gastric ulcer in a 48-year-old Nigerian is reported. This is a rare lesion in the African. The clinico-radiological features that would heighten the clinical suspicion of this condition are highlighted. PMID- 2548313 TI - Case for the panel. Intranuclear paracrystalline structures. PMID- 2548314 TI - Sexually transmitted papillomaviral infections: V. Prophylactic use of topical 5 fluorouracil in refractory infection in the male. AB - The topical administration of 5% fluorouracil (Efudex) was used on 60 males afflicted with recurrent papillomaviral infection, all partners of women with proved HPV disease. The overall response to the medication was 84 percent (total plus partial responses) for those patients afflicted with mixed disease of the shaft, compared with the subclinical and exophytic varieties with 70 and 67 percent response, respectively. Because HPV infections tend to be a field phenomenon involving the entire lower genital tract, this simple form of treatment, that is locally applied with few side effects, appears to be ideal for the management, prophylaxis, and treatment of recurrent disease. PMID- 2548315 TI - Congenital hypomyelinating polyneuropathy in two golden retriever littermates. AB - Serial peripheral nerve biopsies from two golden retriever littermates with chronic neurologic disease were taken for morphologic and morphometric evaluation. Teased nerve preparations were difficult to interpret due to the lightness of myelin staining. Light and electron microscopic findings were characterized by the following: reduced number of myelinated axons, presence of myelinated sheaths inappropriately thin for the caliber of the fiber, poor myelin compaction, increased numbers of Schwann cell nuclei, increased concentration of neurofilaments in myelinated axons, many Schwann cells with voluminous cytoplasm, and increased perineurial collagen. Onion bulb formation was not seen. In contrast to control data, a poor correlation was seen between numbers of myelin lamellae (ML) and axonal circumference (AC). The frequency distribution of ML ranged from 5 to 55 lamellae in affected animals (mean, 28 lamellae) compared to 20 to 140 lamellae in controls (mean, 66 lamellae). The ML/AC ratio was significantly reduced (P less than 0.001) in nerves of affected dogs. Morphometric results indicated that fibers of all calibers were hypomyelinated. PMID- 2548317 TI - Congenital mesoblastic nephroma in a dog: a benign variant of nephroblastoma. PMID- 2548316 TI - Sequential changes in the harderian and exorbital lacrimal glands in Wistar rats infected with sialodacryoadenitis virus. AB - A sequential light and electron microscopic study of the exorbital and Harderian lacrimal glands was done on 2.5- to 15-month-old Wistar rats exposed to sialodacryoadenitis (SDA) virus. Typical coronaviral particles were readily demonstrated in cytoplasmic vesicles of Harderian and exorbital glands examined at 6 days post-inoculation. Lesions were seen in a relatively high percentage of lacrimal glands in infected animals of all ages, with no obvious age-related variations in the incidence and extent of changes. Lesions frequently persisted for a longer interval post-exposure in lacrimal glands than in salivary glands. The persistence of lesions commonly seen in Harderian glands was attributed, at least in part, to the cytotoxic effects of porphyrin-containing secretions released during the acute necrotizing stages of the disease. The persistence of lesions in some lacrimal glands indicates that they are useful tissues for microscopic examination for the retrospective provisional diagnosis of SDA. Persistent lesions also indicate that normal functions of these glands may be compromised for up to several weeks following outbreaks of SDA. PMID- 2548319 TI - Bovine meningoencephalitis from IBR virus. PMID- 2548318 TI - Visceral urate deposits in chicks inoculated with avian nephritis virus. AB - Avian nephritis virus, G-4260 strain, was inoculated orally into one-day-old specific-pathogen-free chicks of two lines. Approximately 20 per cent of the chicks of both lines died with visceral urate deposits from eight to 12 days after infection, and the virus was isolated from the kidneys of the dead chicks. At 14 or 15 days of age the mean liveweight of the surviving infected chicks was approximately 16 per cent less than that of the uninfected control chicks. PMID- 2548320 TI - Aujeszky's disease ELISA: cross-reactions with other herpesvirus antisera. AB - The production of antibodies in pigs to 11 herpesviruses was investigated in relation to their ability to cross-react with Aujeszky's disease virus (suid herpesvirus 1--SHV1). Of the herpesviruses tested only two, sheep herpesvirus (caprine herpesvirus 1) and dog herpesvirus (canid herpesvirus 1), failed to produce homologous virus antibodies. Only the antibodies to bovine herpesvirus 1 (BHV1) produced a cross-reaction by SHV1 enzyme-linked immunosorbent assay (ELISA). No SHV1 neutralizing antibodies were detected in any of the herpesvirus antisera. A cross-reaction with SHV1 by a serum from a pig naturally infected with BHV1 or with any of the other herpesviruses tested was considered unlikely. PMID- 2548321 TI - Ultrastructure and protein A-gold immunolabelling of HRT-18 cells infected with turkey enteric coronavirus. AB - The Minnesota strain of turkey enteric coronavirus (TCV) was propagated in HRT-18 cells, a cell line derived from human rectum adenocarcinoma. A productive non cytopathic infection was established, without a previous adaptation, in these cells as shown by the specific hemagglutinating activity in cell culture supernatants. A post-embedding immunochemical technique, using specific antiserum directed against the original egg-adapted virus and colloidal-gold-labelled protein A as the electron-dense marker, was used for the identification of the virus and related antigens in the cells by electron microscopy. Budding of typical coronavirus particles, through intracytoplasmic membranes and accumulation of complete virus within cytoplasmic vesicles or the lumen of rough endoplasmic reticulum, were the main features of the viral morphogenesis. Late in infection, numerous progeny viral particles were shown at the outer surface of infected cells, but budding could not be demonstrated at this level. Two different types of surface projections were observed on the extracellular particles of this avian coronavirus. These morphological characteristics have been thus far described only for mammalian hemagglutinating coronaviruses. PMID- 2548322 TI - A competitive inhibition ELISA for the differentiation of serum antibodies from pigs infected with transmissible gastroenteritis virus (TGEV) or with the TGEV related porcine respiratory coronavirus. AB - A competitive inhibition ELISA was developed to detect non-neutralizing antibodies to the peplomer protein of transmissible gastroenteritis virus (TGEV) in porcine sera using a monoclonal antibody as an indicator. It was demonstrated that field strains of the TGEV-related porcine respiratory coronavirus (PRCV) did not induce this antibody, whereas the Miller strain and field strains of TGEV did. The sensitivity of the competitive inhibition ELISA appeared to be similar to that of the virus neutralization (VN) test. The test enables differentiation of pigs which were previously infected with TGEV or PRCV and which cannot be distinguished by the classical anti-TGEV neutralization test. The present test is useful for selective serodiagnosis. PMID- 2548323 TI - Redistributive properties of the vesicular stomatitis virus polymerase. AB - The template for transcription of the vesicular stomatitis virus (VSV) genome consists of a negative-strand RNA (approximately 11 kb) tightly associated with approximately 1250 copies of the nucleocapsid or N protein (N-RNA template). The interaction between the virion-associated polymerase and this template was probed with a novel assay using purified N-RNA complexes added to detergent-disrupted uv irradiated standard virions or unirradiated defective interfering (DI) particles. In contrast to the well-known stability of assembled cellular transcription complexes, the VSV polymerase copied exogenously added templates efficiently and yielded products indistinguishable from control virus transcription. Addition of uv-irradiated N-RNA templates to unirradiated virus effectively competed for transcription of endogenous template indicating that most or all of the polymerase can freely redistribute. Furthermore preincubation of virus and added templates at high ionic strength to solubilize L and NS polymerase proteins did not release additional active enzyme for redistribution. Pretranscription of virus also had little or no effect on redistributed activity indicating that polymerase complexes are capable of multiple rounds of synthesis beginning at the 3' end promoter. Unexpectedly, titration with saturating amounts of added N-RNA showed that active polymerase complexes are only in slight excess relative to template in standard or DI particles despite the large surplus of packaged L and NS polypeptides. Moreover, added standard virus templates competed equally well for the redistributing polymerase from DI particles or standard virus indicating no significant polymerase-binding preference for interfering templates. These findings bear important implications regarding mechanisms of VSV transcription and replication. PMID- 2548324 TI - Biological evaluation of glycoproteins mapping to two distinct mRNAs within the BamHI fragment 7 of pseudorabies virus: expression of the coding regions by vaccinia virus. AB - Several glycoproteins from the unique short region of pseudorabies have been identified and characterized. The genes encoding at least four glycoproteins (gp50, gp63, gl, and gX) are located within the BamHI fragment 7 of pseudorabies. S1 nuclease mapping was used to determine that a 2.4-kb mRNA encompasses the coding region for gp50 and gp63 and probably represents a colinear transcript for these proteins. Using the same technique, a 2.8-kb mRNA was found to encode gl. No other mRNAs were found to be encoded on the opposite strand of DNA in this region. Various recombinant vaccinia vectors were made incorporating the coding regions for these two mRNAs. Pseudorabies recombinant vaccinia infected ST cells expressed glycoproteins that co-migrated with the authentic PRV glycoproteins upon polyacrylamide electrophoresis. Intracranial or intraperitoneal inoculation of mice with the recombinant viruses constructed to contain the mRNA coding regions resulted in various degrees of protection from a lethal challenge of pseudorabies virus. PMID- 2548325 TI - The human cytomegalovirus major immediate early promoter can be trans-activated by adenovirus early proteins. AB - We have examined the effect of adenovirus E1 proteins on expression from the immediate early (IE) region of the human cytomegalovirus (HCMV). The major immediate early promoter, responsive to trans-activation during the HCMV lifecycle, is also responsive to E1 a protein encoded by the 13 S message. E1a proteins inhibit SV40 expression through the mechanism of enhancer repression; however, the presence of E1a proteins did not inhibit expression of the IE region of HCMV. The ability of trans-activate the major IE promoter in the presence of a strong enhancer suggests adenovirus can activate transcription of HCMV upon coinfection. E1b proteins increased levels of steady state mRNA transcribed from the IE region. Increases in expression due to E1a and E1b proteins were additive. These results suggest that adenovirus early expression can activate quiescent HCMV sequences. PMID- 2548328 TI - Sequences of VP9 genes from short and supershort rotavirus strains. AB - Segment 10 genes from a short (RV-5, serotype G2) and a supershort (B37, a new G serotype) strain were cloned and their sequences compared to the (corresponding) segment 11 sequences of Wa, SA11, and UK rotaviruses. The determined nucleotide sequences were 817 (RV-5) and 947 (B37) bases in length and showed extensively conserved 5' noncoding and protein coding regions. The major open reading frame codes for a protein of 200 (RV-5) or 198 (B37) amino acids, and the newly proposed second open reading frame can code for a protein of 92 amino acids. Compared to long strain gene segments, the base sequences of the short and supershort strains were found to contain extended, AT-rich 3' noncoding regions which were not significantly homologous to each other, to other parts of the VP9 gene, or to other rotavirus genes that have been sequenced. The function(s) of these 3' regions is not apparent. PMID- 2548327 TI - Rescue of Sendai virus from viral ribonucleoprotein-transfected cells by infection with recombinant vaccinia viruses carrying Sendai virus L and P/C genes. AB - The Sendai virus ribonucleoprotein (RNP) showed only very low plaque-forming titers upon transfection and the virus yields after one-step growth were quite limited. We tried to enhance the Sendai virus yield by supplying the viral L and P/C gene products through vaccinia vectors. A combination of the recombinant vaccinia viruses carrying the L gene (Vac-HL) and the P/C gene (Vac-HPC), both of which were driven by the promoter of the vaccinia virus 7.5K protein gene, enhanced the yield only a little whereas another combination of Vac-HLd7.5, the L gene insert of which was driven by the promoter of the vaccinia virus thymidine kinase gene in place of the 7.5K promoter, and Vac-HPC greatly enhanced the Sendai virus yield. This seemed to correlate with the fact that the Vac-HL interfered with Sendai virus growth markedly while the Vac-HLd7.5 did not. These results strongly suggest that the L and P/C gene products act in cooperation as the RNA polymerase, and overproduction of the L protein is inhibitory for Sendai virus growth. This system seems to be of value as a tool for analyzing the functions of L and P/C genes of Sendai virus. PMID- 2548326 TI - Expression of Epstein-Barr virus nuclear antigens 3, 4, and 6 are altered in cell lines containing B-type virus. AB - A high proportion of HIV-positive sera were found to react with 130- and 180-kDa antigens which were present in the Jijoye cell line. The majority of the HIV positive sera which detected these antigens also contained antibodies to Epstein Barr virus (EBV) nuclear antigen 2B (EBNA2B) suggesting a relationship between B type EBV strains and the expression of the 130K/180K antigens. Cell lines were established by infection of B lymphocytes with different A- and B-type strains of EBV. Incubation of these lines with sera from individuals infected with either A type or B-type EBV strains demonstrated that the 130K and 180K antigens were only expressed by cell lines containing B-type virus. Sera from individuals infected with A-type EBV did not react with the 180K antigen in any cell lines but could detect EBNAs 3, 4, and 6 antigens in the A-type cell lines. Restriction enzyme analysis of the BamHI E region of the EBV genome revealed marked differences between the A and B types of the virus. These results demonstrate that expression of antigens encoded from the BamHI E region of EBV (EBNAs 3, 4, and 6) are altered in cell lines transformed by B-type strains of EBV. PMID- 2548329 TI - Stably expressed FIPV peplomer protein induces cell fusion and elicits neutralizing antibodies in mice. AB - We have established bovine papilloma virus (BPV)-transformed mouse C127 cell lines that synthesize the peplomer protein of the feline infectious peritonitis virus (FIPV) strain 79-1146. For this purpose, a new cassette expression vector pHSL, which carries the Drosophila HSp70 promotor and the polyadenylation signal of the Moloney murine leukemia virus long terminal repeat, was constructed. Cocultivation of the BPV-transformed cell lines with FIPV-permissive feline fcwf D cells resulted in polykaryocyte formation. Since it depended on the presence of fcwf-D cells, binding of E2 to the cell receptor may be required for membrane fusion. E2 was synthesized as a core-glycosylated protein of 180K which was only slowly transported from the endoplasmic reticulum to the medial Golgi: of the E2 molecules labeled during a 1-hr pulse about half was still completely sensitive to endoglycosidase H after a 2-hr chase, while the remaining E2 had been chased into multiple, partially endoglycosidase H-resistant forms. Immunofluorescence studies also indicated that most E2 was retained intracellularly. Mice immunized with whole lysates of the transformed cells produced FIPV-neutralizing antibodies as shown by plaque reduction. PMID- 2548330 TI - Modification of foot-and-mouth disease virus after serial passages in the presence of antiviral polyclonal sera. AB - Foot-and-mouth disease virus (FMDV) shows a remarkable antigenic variability. Like other RNA viruses, this virus has a high rate of mutation. It has been proposed that selection exerted by the host's antibodies could play a major role in the rapid evolution of FMDV. The present work reports the selection of FMDV antibody-resistant populations (Nr), after serial passages of cloned FMDV A24 Cruzeiro strain on secondary monolayers of bovine fetal kidney cells in the presence of subneutralizing antiviral polyclonal sera (APS). After a limited number of passages under selective pressure, the virus population showed the following characteristics: (1) increased resistance to neutralization by APS; (2) altered electrophoretic mobility of structural viral proteins (VP1); (3) remarkable plaque size reduction, (4) a pronounced thermosensitivity (ts); and (5) decreased pathogenicity for mice, in both uncloned and cloned small plaque size populations. This indicates that FMDV populations under antibody pressure in vitro, have acquired, in addition to expected characteristics of natural FMDV variants (resistance to neutralization and altered viral structural proteins), phenotypic markers which correspond to attenuated, less virulent variants. PMID- 2548331 TI - Regulated transcription of herpes simplex virus immediate-early genes in neuroblastoma cells. AB - C1300 neuroblastoma cells are nonpermissive for infection with herpes simplex virus but can be rendered permissive by pretreatment with sodium butyrate. This increased permissivity which is specific for HSV is caused by increased transcription of the viral immediate-early genes following infection of butyrate treated cells and can be observed for at least 24 hr following withdrawal of butyrate. The use of C1300 cells as a model system for studying the regulation of immediate-early gene expression in neuronal cells in vitro and its possible relevance to the study of the processes regulating latent infection in vivo is discussed. PMID- 2548332 TI - Trypsin sensitivity of several human rhinovirus serotypes in their low pH-induced conformation. AB - Five serotypes of human rhinovirus (HRV) were examined for sensitivity to trypsin at physiological pH, HRV1A, HRV2, and HRV14 were found to be resistant whereas in serotypes HRV49 and HRV89 degradation of VP2 was observed. However, exposure to low pH followed by neutralization, a treatment which causes irreversible conformational changes in the capsid, led to rapid cleavage by trypsin of VP1 in HRV1A, HRV2, and HRV49 at defined sites followed by degradation of VP2. In the case of HRV2, the cleavage site in VP1 was determined by direct protein sequencing and was shown to occur between Arg260 and Thr261, close to the C terminus. HRV49 behaves similarly to HRV2 as expected from extensive sequence similarity in this region, whereas VP1 in HRV1A is most probably cleaved at a site closer to the C-terminus than that in HRV2. Although HRV14 contains the same amino acid pair present in HRV2 and HRV49, it was not cleaved under these conditions. HRV89, which lacks a basic residue at the corresponding position, was also insensitive. Examination of the cleavage site on the three-dimensional structural map of native HRV2 reveals that it is most probably buried inside the capsid and thus not accessible. Structural rearrangements of the viral capsid are thus necessary to account for the cleavage observed after low pH treatment. PMID- 2548333 TI - Structural and functional characterization of mutants of the bovine leukemia virus transactivator protein p34. AB - Mutants of the bovine leukemia virus (BLV) transactivator protein (tat, tax, p34, the XLOR gene product) were constructed by site-directed deletions, in-phase linker insertions, or fragment replacements (swapping) among BLV variants. The mutant constructs were transfected into cos cells and transiently expressed. Western blot analysis using a mixture of monoclonal antibodies to wild-type p34 revealed the presence of mutated XLOR gene products in all the mutants tested. The transactivating activity of 11 tax mutants containing site-directed deletions and in-phase linker insertions was completely abolished. Only the swapping mutant tested, a hybrid between two BLV variants, transactivated LTR-directed gene expression at wild-type levels. These data illustrate the narrow range of structural variations that allow full activity of the BLV tax product and suggest that the present molecular structure of the transactivator protein results from heavy evolutionary constraints. PMID- 2548334 TI - Glycoprotein gIII deletion mutants of pseudorabies virus are impaired in virus entry. AB - gIII is one of the major structural glycoproteins of pseudorabies virus (PrV). Though nonessential for replication in cell culture, it plays a prominent role in virus adsorption and virus release from infected cells. In this study the effect of inactivation of gIII on virus uptake into cells was investigated using isogenic PrV glycoprotein mutants. Kinetic analyses demonstrated that deletion of the gIII gene severely impaired entry of PrV into cells, whereas inactivation of the genes coding for nonessential glycoproteins gI, gp63, or gX had no influence on the rate of virus penetration. Loss of gIII is therefore associated with a reduced rate of virus penetration indicating a gIII-independent way of virus entry which is significantly slower than the gIII-dependent penetration process. PMID- 2548335 TI - Persistent viral DNA synthesis associated with an avian osteopetrosis-inducing virus. AB - Bone and red blood cell DNA was obtained from MAV-2(O) infected, osteopetrotic chickens and analyzed for the presence of integrated and unintegrated viral DNA. Results indicate that unintegrated MAV-2(O) DNA did not appear until after osteopetrotic lesions were well established. These observations lead us to conclude that, unlike some retroviral diseases, unintegrated viral DNA may not play a significant role in the pathogenesis of MAV-2(O) osteopetrosis. In addition, there was no evidence of a clonally derived tumor in the bone. PMID- 2548336 TI - Detection of a trypsin-like serine protease domain in flaviviruses and pestiviruses. AB - We propose through a sequence and structural-pattern analysis that a protein domain of undefined function encoded by the enveloped RNA flavi- and pestivruses is a Ser active-center enzyme related to the cellular trypsin family. A further homology is emphasized with the group of (Cys active-center) viral proteases encoded by nonenveloped RNA viruses of the picorna-, como-, nepo-, and potyvirus classes. Structural modeling of the putative flaviviral protease domain suggests amino acids that are crucial for catalytic activity and substrate binding. PMID- 2548337 TI - Transcriptional activation is not responsible for increased levels of autonomously expressed simian virus 40 T-antigen in herpes simplex virus-infected cells. AB - Herpes simplex virus type 1 (HSV-1) superinfection of CV-1 cells weakly transactivated a plasmid-borne metallothionein 1 (MT-1) promoter, but activated the expression of a marker gene controlled by an authentic HSV-1 promoter to a high level. In contrast, CMT-3 cells, which are CV-1 cells stably transformed with the simian virus 40 (SV40) large T-antigen (T-Ag) gene controlled by the MT 1 promoter, contained high levels of T-Ag following HSV-1 superinfection, but only if cells were preincubated in the presence of heavy-metal ions. This T-Ag was functional in that it could mediate the increase in copy number of a marker plasmid containing the SV40 origin of DNA replication. Pulse and continuous labeling of preinduced CMT-3 cells showed that T-Ag expression was not induced by HSV-1; but rather, HSV-1 superinfection resulted in the stabilization of pre existing protein. PMID- 2548338 TI - Enhancement of herpes simplex virus type 2 (HSV-2) DNA synthesis in infected cells that constitutively express the BglII-N region of the HSV-2 genome. AB - The BglII-N fragment of the herpes simplex virus type-2 (HSV-2) genome encodes one of two known transforming regions of this DNA virus. In this study, we report the derivation of HeLa S3 cells (2DC4) that stably express the HSV-2 BglII-N region, including the small subunit of HSV-2 ribonucleotide reductase (RR). Superinfection of the 2DC4 cells with wild-type HSV-2 resulted in the efficient induction of HSV-2-encoded ICP10, DNA polymerase, and thymidine kinase. The amount of HSV-2 DNA synthesis in 8-hr HSV-2-infected 2DC4 cells was enhanced 2.6 +/- 0.6-fold relative to infected control cells. Furthermore, the replication kinetics of HSV-2 DNA in 2DC4 cells were accelerated relative to HeLa S3 cells; HSV-2 DNA synthesis was detectable as early as 3 hr postinfection in 2DC4 cells as compared to 6 hr postinfection in HeLa S3 cells. These results suggest that the BglII-N region of HSV-2 encodes function(s) that activate the viral DNA synthesis apparatus and that this activation could relate to the transforming ability of this DNA region. PMID- 2548339 TI - Sequences of capsid protein VP1 of two type A foot-and-mouth disease viruses. AB - We have sequenced the nucleotides of the regions that encode the capsid protein VP1 of the foot-and-mouth disease viruses (FMDV) A5Bernbeuren/1984 and A Iran/1987. Amino acid sequences and secondary protein structures are provided. Both proteins consist of 212 amino acids. The sequences and secondary structures are compared to those of FMDV A22/CCCP/64, a strain previously endemic in the Near East. Nucleotide divergency among the three sequences is highest for FMDV A5Bernbeuren/1984 (18% compared to 13% for each other case). Thirty amino acid divergencies are observed between A22/CCCP/64 and A5 Bernbeuren/1984 or A Iran/1987, whereas the latter two differ by 27 residues. The secondary structures of all three proteins are different. A Iran/1987 is considered to belong to a thus far unknown subtype. PMID- 2548340 TI - Nucleotide sequence between the peplomer and matrix protein genes of the porcine transmissible gastroenteritis coronavirus identifies three large open reading frames. PMID- 2548341 TI - [The calcium signal in the physiology of the secretory process. II. The effect of extracellular and intracellular calcium on secretory mechanisms]. AB - Investigations of the influence of the extra- and intracellular calcium source on secretory processes helps the understanding of the physiology and pathophysiology of secretion. The ratio of the two calcium sources in the activation of secretion depends on the type of secretory cell, on the character of the secretagogue stimulus and the phase of the secretory process. The use of intracellular sources of calcium in the early stage of secretion explains the lack of an inhibitory action of calcium current blockers on the function of some hormonal systems in short-term studies, e.g. the tyrotropic and somatotropic system, or on adrenocortical secretion. Conversely inhibition of some hormonal functions in the curse of prolonged inhibition of calcium currents indicates the importance of the inflow of calcium ions from the extracellular space into the cytosol during long term regulation of secretion. The author gives an account of the results of investigations which by means of calcium ionophores or calcium current antagonists provide evidence of the importance of an extracellular calcium source in the control of secretion of some systems. PMID- 2548342 TI - [The spatial interrelationships between influenza virus hemagglutinin and the lipid phase in the liposomal membrane]. AB - A scheme has been proposed demonstrating the location of tryptophan residues of hemagglutinin molecule in relation to the middle of the lipid layer 1.2 nm thick with a fluorescent probe pyrene. In the immediate proximity to it, one tryptophanyl of protein molecule is located in a hydrophobic "pocket". At a distance of 2.85 nm from the middle of the lipid zone 3 tryptophanyls are located and the remaining five at a distance over 3.6 nm. After treatment with proteolytic enzyme bromelin of the liposomes with hemagglutinin incorporated into their bilayer, the hydrophobic "anchor" of protein molecule contains one tryptophanyl which is raised by 0.3 nm and its hydrophobic environment is changed. PMID- 2548343 TI - [Comparative study of the hemolytic activity of ortho- and paramyxoviruses]. AB - A comparative study of hemolytic activity of influenza type A, B, and C viruses, human parainfluenza type 3, and Sendai virus showed the pattern of pH-dependence and the nature of the curve to differ not only for different viruses under study but also for different erythrocyte species. Studies of virus-induced hemolysis of influenza C virus demonstrated that, depending on the erythrocyte species used, it had common properties both with influenza types A and B viruses and with paramyxoviruses. PMID- 2548344 TI - [Serological proof of the existence of a variety of non-A, non-B hepatitis with a fecal-oral transmission mechanism]. AB - An outbreak of hepatitis in one of Siberian regions mainly among adults of 19-35 years of age was studied. The epidemiological studies showed the infection to spread by the fecal-oral mechanism of transmission. In 90% of the patients the disease was mild, in 10% of moderate severity; no severe forms or fatal outcomes were observed. This refers also to pregnant women irrespective of the term of pregnancy. Among 78 subjects examined, antibody to hepatitis A virus of the IgM class (anti-HAV-IgM) were detected in 14%, HBsAg in 6%. No rise in titer of total anti-HAV in the convalescent period (within 3-6 months) was observed. Immune electron microscopy studies of fecal extracts from patients revealed immune complexes of virus particles 32-35 nm in size which were formed only with sera from patients in the acute stage and convalescents from this outbreak (to a lower degree). Negative results were obtained with sera containing anti-HAV and antibody to hepatitis non-A-non-B virus (HnAnB) with fecal-oral transmission mode from India, Central Asia, and Afghanistan. These data attest to the distinct nature of this causative agent among HnAnB viruses with the fecal-oral transmission mechanism. PMID- 2548346 TI - [A comparison of laboratory diagnostic methods for rotavirus gastroenteritis in a practical laboratory]. PMID- 2548345 TI - [Immunoenzyme analysis of adenovirus hexon: serodiagnosis in human eye diseases]. AB - The enzyme-linked immunosorbent assay (ELISA) was adapted for detection of IgG antibodies against adenovirus hexon in the sera and tears of patients with eye diseases. The ELISA procedure is more sensitive than complement fixation test for the detection of antihexon antibodies; it allows the seroconversion to be seen in the course of infection and convalescent period. The ELISA procedure has been used for testing specimens from a group of 66 patients with keratoconjunctivitis of adenoviral etiology. PMID- 2548347 TI - [Use of the indirect hemagglutination reaction for the diagnosis of rotavirus gastroenteritis]. PMID- 2548348 TI - Bombesin-like peptides as growth factors. AB - Bombesin-like peptides (BN-LP) are involved in the regulation of many important functions, including sensory transmission, regulation of central autonomic pathways, thermoregulation, pituitary, gastric and pancreatic secretion, food intake and satiety. They also stimulate cellular proliferation in a developmental and tissue-specific manner. Their role in pathogenesis appears to be related to their properties as growth factors, especially in the lung, where BN-LP can induce growth of normal and neoplastic epithelial cells. The formulated hypothesis of autocrine control of small cell lung cancer growth by BN-LP will be tested using specific synthetic bombesin antagonists. PMID- 2548350 TI - [Human herpesvirus 6 (HHV-6): incidence in healthy blood donors and in patients with acute infections caused by other herpes viruses]. AB - Antibodies to HHV-6 were detected by immunofluorescence in 8.04% of 460 healthy blood donors in West Austria. Testing sera from patients with acute or reactivated infections with other herpesviruses, such as cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex virus (HSV) and varicella zoster virus (VZV) we observed a remarkably higher prevalence of antibodies to HHV-6 in patients with CMV infections (75%) and also in patients with EBV infections (50%). Patients with HSV and VZV infections were positive for HHV 6 in 12.5% and 6.6% of cases, respectively. Many of the patients with CMV infection were transplant recipients. The high incidence of positive HH 6 serology in these patients could be due to new infection by HHV 6 or to the reactivation of a previous infection with HHV 6 by means of allogenic cell stimulation. Furthermore, preliminary results from our laboratory point to a serological cross reaction between HHV 6 and CMV, which may also contribute to this result. PMID- 2548349 TI - [The immunology of aging: immunoregulatory role of lipoproteins]. AB - This report deals with immunoregulation by lipoproteins and with age-related changes in immune function. It was investigated whether a diminished immune response during ageing could be caused by a disturbed interaction of lipoproteins with cells of the immune system. The aim of these investigations was to show whether changes in the immune response in the elderly were really age-related, or attributable to other factors (i.e. medication, life style) not related to ageing per se. Therefore, admission criteria for blood donors to take part in these studies of the ageing human immune system were set up by using the Senieur protocol (EURAGE). An immunoregulatory influence of lipoproteins from different density fractions-Low Density Lipoproteins (LDL), High Density Lipoproteins (HDL) and Very Low Density Lipoproteins (VLDL) on the in vitro mitogen response of human peripheral blood lymphocytes (PBL) was shown. Lymphocytes can take up lipoproteins by receptor-mediated (specific binding) and non receptor-mediated (receptor-independent exchange) mechanisms. In this context a HDL-binding site/receptor was demonstrated and characterised here for the first time on human T-cells. Furthermore, interactions of lipoproteins with lymphocytes were investigated in the elderly and compared with young control groups. The increased LDL uptake during ageing was found to correlate with an increased HDL uptake. The age-related diminished immune response correlated with a decrease in plasma membrane fluidity of human PBL, but not with an enhanced lipoprotein uptake. The Senieur protocol in this present form, used for classifying blood donors, had no influence on distribution and statistical processing of the obtained sets of data, indicating that this protocol needs further modifications. PMID- 2548351 TI - Detection of free radicals as a consequence of dog tracheal epithelial cellular xenobiotic metabolism. AB - 1. Spin-trapping techniques have been used to examine the metabolism of three xenobiotics known to produce free radicals during their metabolism. Reaction with oxygen generated superoxide, the location of which was dependent upon the xenobiotic. 2. Paraquat was metabolized by dog trachea epithelial cells under anaerobiosis to the paraquat free radical, some of which diffused into the extracellular milieu. With the addition of oxygen, superoxide was spin-trapped both intra- and extracellularly. 3. When menadione was metabolized by epithelial cells, superoxide was spin-trapped within the cell and in the surrounding media. However, in this case, extracellular superoxide arose as the result of the disproportionation reaction of menadione and menadiol, resulting from DT diaphorase reduction of menadione followed by diffusion into extracellular space, to give the menadione semiquinone. Reduction of oxygen resulted in formation of superoxide. 4. For nitrazepam, only intracellular superoxide was generated, resulting from the one-electron reduction of this drug to its corresponding nitro anion free radical. Reaction with oxygen produced superoxide. PMID- 2548352 TI - Nonoxynol 9 in herpes. PMID- 2548353 TI - [Local pigment stimulation by alpha-melanocyte stimulating hormone in the human: intracutaneous injections without and with ultraviolet irradiation]. AB - Repeated intracutaneous injections of alpha-MSH failed to stimulate local pigmentation in healthy persons. Only additional application of UV irradiation resulted in a distinct brown discoloration of the skin at the site of injection. PMID- 2548354 TI - Efficient medium for impingement and storage of enveloped viruses. AB - Airborne infections with pathogenic viruses play an important role in the transmission of diseases amongst men and animals. We compared several media intended for impingement of viruses from virus-contaminated air and for their preserving effect for two enveloped viruses. Sindbis (SINV) and vesicular stomatitis virus (VSV), members of the families Toga- and Rhabdoviridae, respectively, were chosen as indicator agents. Amongst the media tested, a sampling fluid consisting of phosphate buffered saline, pH 7.2, 0.5% bovine serum albumin, 0.5% gelatine (PBSplus) was most efficient to minimize the sampling stress during impingement and to preserve the infectivity SINV and VSV under stringent conditions at 37 degrees C. About 50% of virus infectivity was recovered 15.7 or 30 hours, respectively, after the beginning of storage. Thus the recommended medium is also suitable for shipment and storage of diagnostic virus samples. PMID- 2548355 TI - [The occurrence of Escherichia coli, Aeromonas hydrophila, Plesiomonas shigelloides and Clostridium perfringens in the intestinal flora of gray herons (Ardea cinerea)]. AB - The flora of the large intestine of 92 grey herons was examined for the frequency of aerobic and microaerobic growing bacteria. Clostridium perfringens, Aeromonas hydrophila, Plesiomonas shigelloides and E. coli were isolated from 55%, 48%, 14% and 35% of the birds, respectively. It could be demonstrated that the findings of these bacteria in the intestinal flora are depending on the age of the birds. The percentage of carriers of Clostridium perfringens, Aeromonas hydrophila and Plesiomonas shigelloides was highest in nestlings younger than 18 days, less high in older nestlings and lowest in adult grey herons. Contrary to those bacteria, E. coli was found more often in the intestinal flora at increasing age of the birds. Salmonella spp. were isolated from 6 birds. Two birds yielded positive for Yersinia enterocolitica and Campylobacter spp., respectively. Other aerobic and microaerobic bacteria play a less significant role as part of the intestinal flora. PMID- 2548356 TI - [The detection and identification of poxvirus in domestic cats]. AB - Two cases in Southern Germany of pox in cats could be diagnosed virologically. A cowpox-like virus was isolated, its biological and serological characteristics defined. The source of infection could not be traced. The contraindication of treatment with corticosteroids and the pathogenicity of the orthopoxvirus demonstrated here for humans is pointed out. PMID- 2548357 TI - Experimental transmission and electron microscopic demonstration of the virus of haemorrhagic disease of rabbits in Czechoslovakia. AB - Intramuscular administration of the filtrate of organ suspensions, prepared from a dead rabbit, killed 62.9% of inoculated rabbits within 1 to 5 days, while 93.3% died after intranasal administration of the same inoculum. The virus survived freeze-drying and was resistant to treatment with 0.4% formaldehyde when incubated at 37 degrees C for 1 hour and 4 degrees C for the subsequent 12 hours, but lost its infectivity when the treatment was prolonged to 3 hours at 37 degrees C and 3 days at room temperature. Its infectivity was also inhibited by reconvalescent serum. The virus could not be detected after 3 passages in primary rabbit kidney cell cultures. Electron microscopy of negatively stained preparations demonstrated icosahedral virus particles with a diameter of 29 to 33 nm without an envelope. Accurate morphological classification has not yet been completed. Incubation with a reconvalescent serum, diluted 1:20 or 1:40, resulted in the formation of immune complexes, detectable by electron microscopy. PMID- 2548358 TI - Injection of catalytic subunit of cyclic AMP-dependent protein kinase speeds up the development of junctional communication in the embryo. AB - Using the time course of high capacity junctional communication development in endoderm of the early Xenopus embryo as the measure the effect of cAMP and the catalytic subunit of cAMP-dependent protein kinase on this development has been investigated. It is found that while cAMP has no effect on this development the catalytic subunit strongly speeds it up. The discrepancy may reflect the high concentration of cAMP and low concentration of effective cAMP-dependent protein kinase in endoderm cells of the early Xenopus embryo. PMID- 2548359 TI - [Changes of Golgi complex during the induced differentiation of human gastric adenocarcinoma cells in vitro]. AB - Golgi complex is a central link related to each kind of organella in cellular metabolic process, its morphological changes are often concerned with the cell differentiation and functional state. The Golgi complexes in tumour cells have the characteristics of poor development and non-typical structure, and closely related to their pathological differential degree. For this reason, having based on appraising the differential effect of human gastric adenocarcinoma cell line MGc 80-3 induced by dBcAMP in vitro, we had made a systematic observation on the changes of Golgi complex during the malignant phenotypical reversion of MGc 80-3 cells, in order to inquire into the relationship between the structural and functional changes of Golgi complex and the malignant phenotypic reversion of cancer cells. It was revealed by ultrathin sectioning and freeze-etching electron microscopy that, in MGc 80-3 cells, there were a few Golgi complexes, their volume was small, the amounts of saccule were few, they arranged irregularly, expanded and inflated, and the intramembranous particles on saccule were rare and not well-distributed. It displayed a non-developmental and non-typical structural state of Golgi complex. But after induced treatment with dBcAMP, the Golgi complexes had grown in number and distributed concentrated, , their volume enlarged, the saccules increased and arranged regularly, and the intermembranous particles on saccules were plentiful and well-distributed. They had restored a well-developed and typical structure of Golgi complex similar to that of the primary culture cell of human normal gastric mucous membrane. It showed that Golgi complex had further changed into a quite developed state during the induced differentiation. This alteration not only inhibited the malignant secretory activity of gastric carcinoma cells but also played a certain regulative role in the changes of the surface components of cancer cells. The structural and functional changes of Golgi complex were considered to be an important expression in the malignant phenotypical reversion of cancer cells, it had an important influence on the differentiation of cancer cells from malignant to normal direction. PMID- 2548360 TI - Genetic relatedness among Mycobacterium tuberculosis and M. bovis. AB - Total DNA from two slowly-growing pathogenic mycobacterial species propagated in vitro was isolated, digested with each of 34 restriction endonucleases and analyzed by agarose gel electrophoresis. The most resolved patterns for M. tuberculosis (ATCC 27294) and for M. bovis (ATCC 19210) were obtained respectively using (BamHI, DraI, ClaI, EcoRI, EcoRV, HindIII, HpaI, SalI, SmaI, XbaI and XmaI). The patterns produced for these strains were reproducible and distinguishable from each other respectively using (HindIII, DraI, EcoRI, MboI, Sau3AI and AvaI). However, with several enzymes (SalI, AsuI, Sau96I, MspI and HpaII) the patterns for M. tuberculosis and M. bovis were similar. Evidence was obtained for the presence of dam and dcmI methylations in the DNA of each mycobacterial species respectively using (MboI, Sau3AI, EcoRII, BstNI, Sau96I and AsuI). PMID- 2548361 TI - Insertion elements in mycobacteria. AB - We have isolated and characterised a repetitive element from the genome of Mycobacterium paratuberculosis. This repetitive element shows many features characteristic of a bacterial insertion element. PMID- 2548362 TI - Studies on adenylate kinase (ATP:AMP phosphotransferase) of Mycobacterium marinum (ATCC 927). AB - The enzyme adenylate kinase (ATP:AMP phosphotransferase) was purified as described previously [Biochim. Biophys. Acta 869: 350 (1986)] with an additional step involving affinity chromatography on Cibacron Blue. The molecular weight of the final enzyme preparation was estimated to be 22,500 on polyacrylamide-gel electrophoresis under denaturing conditions. The preliminary amino acid analysis indicated the presence of two histidine residues. Photooxidation in the presence of methylene blue caused complete inactivation of the enzyme, but the loss of activity could be prevented by the addition of ATP, AMP or adenosine-(5') pentaphospho-(5')-adenosine, indicating that at least one histidine residue is involved at the active site. A circular dichroic study indicated that the enzyme consists of 24% alpha-helix, 30% beta-structure, and 46% random coil. The bacterial cells induced with antimycin A and light (particularly with the former) appeared to have somewhat increased adenylate kinase activity, although the Km and Vmax values were unchanged. PMID- 2548363 TI - Isoenzymes as tools to discriminate various subdivisions in the Mycobacterium fortuitum complex. AB - Methods for the characterization of catalase, peroxidase, beta-glucosidase, esterase, and beta-lactamase mycobacterial isoenzymes were described. These methods were applied to examine strains of the M. fortuitum complex. M. fortuitum, M. peregrinum, M. chelonae- M. abscessus and an unnamed species had distinct isoenzyme profiles. M. chelonae and M. abscessus could not be satisfactorily differentiated using the described methods. PMID- 2548364 TI - Isoenzymes profiles in slowly-growing and difficult-to-grow mycobacteria. AB - The cell-free extracts of 13 slowly-growing mycobacteria were run on polyacrilamide gels (Page) and the various protein bands obtained were tested for peroxidase and catalase enzyme activities. The results obtained were compared to those obtained with M. fortuitum and M. chelonae. Based only on a limited numbers of strains employed, it is suggested that these isoenzyme patterns may permit a better separation of "Wood-pigeon" mycobacteria from both M. avium and M. paratuberculosis and also give distinct profiles for other species used. These results further suggest the potential of isoenzymes as taxonomic markers. PMID- 2548366 TI - Nasal mucociliary clearance in Sjogren's syndrome. Dissociation in flow between sol and gel layers. AB - Nasal mucociliary clearance was measured with two methods in 8 patients with Sjogren's syndrome and in 6 normal subjects. The movement of two different tracers placed 1.5 cm posterior to the inferior turbinate tip was measured respectively. The transport rate of a 500 microns anion resin particle tagged with 99mTc was measured. The clearance of 10 microliters saline labelled with 99mTc was monitored and the clearance rate was calculated. Whereas the measurement with the particle method revealed the lowered transport rate in Sjogren's syndrome, measurement with the saline method did not reveal any difference in clearance rate between the two groups. Since the former method measures the transport of particle in the gel phase (the outer mucous layer) and the latter measures the clearance of both gel and sol (periciliary fluid) layers, it is postulated that there is a dissociation of flow between sol and gel layers in Sjogren's syndrome. PMID- 2548365 TI - Embryonal neuroepithelial tumors induced by human adenovirus type 12 in rodents. 2. Tumor induction in the central nervous system. AB - Intracranial inoculation with human adenovirus type 12 (Ad12) induced tumors multicentrically in the brain and spinal cord of 37.2% of hamsters, 30.2% of mice, and in the brains of 91.0% of rats. Brain tumors developed preferentially at the olfactory bulb, lateral ventricular horns, tapetum region, and ventral and dorso-caudal aspects of the fourth ventricle. In the spinal cord, tumor developed on the dorsal aspect and, in hamsters, at the root of the cauda equina. Microtumors were found almost invariably in the subependymal areas and occasionally in the leptomeninges. The histological and ultrastructural features indicated extremely undifferentiated neoplasms analogous with the intraperitoneal tumors described in the companion report. Closely packed small polygonal or tadpole-shaped tumor cells resembled the subependymal cell remnants of normal perinatal brains. Divergent differentiation consisted in an intermingling of a fascicular or palisading arrangement of spongioblastic cells, of incomplete perivascular pseudorosettes and of neuroblastic (Homer Wright type) rosettes. Neither distinct neuronal nor neurogial fibrils were demonstrated. True ependymoblastomatous and medulloepitheliomatous rosettes were rarely encountered. These results indicate that Ad12-induced tumors in the central nervous system are of embryonal neuroectodermal origin and with limited differentiation, leading to divergent phenotypes corresponding to medulloblastoma, neuroblastoma, primitive spongioblastoma, ependymoblastoma and, rarely, medulloepithelioma. PMID- 2548367 TI - [Clinical and biological considerations on adenocystic carcinoma of the head]. AB - The purpose of the present work was to define some aspects (i.e. the influence of histological grading on prognosis and the possibility of an eventual hormone dependency) of adenoidocystic carcinoma, a highly controversial form of carcinoma still open to debate. A study is thus reported of 33 cases of adenoidocystic carcinoma of the head and neck seen from 1965-1985 at the E.N.T. Clinic of Ferrara. The individual histological samples were seen over a period of time by a single histopathologist who was not informed as to the patients' clinical progression. In this manner the diagnostic criteria was kept constant. Tumoral grading was then performed on the basis of the Marsh and Allen classification. The overall actuarial survival curves are presented according to site and grading. Besides confirming the already known data regarding the decrease in survival rate well beyond the fifth year, local recurrences and the frequency of lymph node and secondary metastases, the results indicate a direct relationship between grading and prognosis. In fact, overall survival decreases for the less differentiated forms while the frequency of local recurrences increases as does that of lymph node and systemic metastases. Furthermore, in 63.3% of the female patients a genital hormone dependent pathology was noted prior to the appearance of a salivary gland tumor. These observations would suggest that hormones may play a role in the genesis of adenoidocystic carcinoma. PMID- 2548368 TI - [Ultrastructural demonstration of the human papilloma virus (HPV) in oral proliferative lesions]. AB - The Human Papilloma Virus (HPV) has been found in a series of benign proliferative lesions of the skin and mucosa. The virus has also been found in verrucous laryngeal carcinoma and carcinomas of the oral cavity and other organs. DNA hybridization techniques have made it possible to classify 51 types of HPVs, some appearing to be associated with specific lesions. In order to study the intracellular distribution of HPVs, an ultrastructural morphological analysis was performed with an electron microscope on 10 specimens taken from 5 patients. The specimens were obtained through large excisional biopsy, histologically classified as "fibropapilloma". For each patient specimens were taken from the clinically evident lesion and from the surrounding clinically normal mucosa. The specimens were fixed in glyceraldehyde, washed in a cacodylate buffer, post-fixed in potassium ferrocyanide reduced-osmium tetroxide, stained with uranyl acetate and included in EPON 812. The tissue was subjected to amylase digestion prior to electron microscope examination. A great number of viral particles were found in both the nucleus and the cytoplasm, without forming crystal array structures as typically described for the verruca vulgaris virus (HPV-2). No significant differences could be found between the cells derived from the clinical lesion and those derived from the surrounding mucosa. The passage of viral particles from infected to as yet uninfected cells through the intercellular space could be seen. The high intracytoplasmatic presence of the virus, and its clear abundance in those cells surrounding the clinical lesion, is felt to be of special interest. PMID- 2548369 TI - Effects of aldosterone and betamethasone on Na-K-ATPase activity in the juxtamedullary proximal convoluted tubules of mice. AB - An attempt was made to determine whether or not the steroid hormones, aldosterone and betamethasone, increased the Na-K-ATPase activity and the surface density of baso-lateral cell membranes in the juxtamedullary proximal convoluted tubules of young and adult mice. The results showed that aldosterone and betamethasone increase the enzyme activity in young mice but not in adult mice. However, the surface density of the baso-lateral cell membranes did not increase either in the young or in the adult mice. Further, the induction ratio for the Na-K-ATPase activity did not differ between the two hormone-treated groups in young mice. The results suggest that a difference in hormone sensitivity exists between the proximal convoluted tubular cells of young mice and those of adult mice. PMID- 2548370 TI - Endocrine responses to growth hormone-releasing hormone, thyrotropin-releasing hormone and corticotropin-releasing hormone in depression. AB - To explore and to compare hypothalamic-pituitary-somatotropic (HPS), hypothalamic pituitary-thyroid (HPT) and hypothalamic-pituitary-adrenocortical (HPA) axis function in depression, 30 subjects (15 patients with a major depressive episode and individually matched controls) received 50 micrograms growth hormone releasing hormone-44 amide at 9:00, 200 micrograms thyrotropin-releasing hormone (TRH) at 9:00 and 100 micrograms human corticotropin-releasing hormone (CRH) at 18:00 on consecutive days as an i.v. bolus dose. Compared with controls, depressed patients showed blunted growth hormone (GH) responses to GHRH, decreased TRH-induced thyrotropin (TSH) release and reduced corticotropin (ACTH) but normal cortisol secretion following CRH. ACTH secretion following CRH and TRH induced TSH release were positively correlated across depressed patients and controls but no significant correlations between GH responses to GHRH and TRH induced TSH release or ACTH and cortisol secretion following CRH administration were demonstrated. Our findings suggest that altered HPT and HPA axis function associated with depression are triggered by factors that are at least partly different from those that cause HPS system dysfunction. We conclude that the pathophysiological process resulting in aberrant neuroendocrine secretory dynamics associated with depression may primarily occur at a suprapituitary site, and that HPS, HPT and HPA axis dysfunction may be precipitated by complex central and peripheral regulatory mechanisms involving largely independent factors. PMID- 2548371 TI - Hypothalamic-pituitary-adrenal and -thyroid axis dysfunctions and decrements in the availability of L-tryptophan as biological markers of suicidal ideation in major depressed females. AB - Several neurochemical correlates of suicide were recently detected. Some authors found increased disorders in the hypothalamic-pituitary-adrenal (HPA) and thyroid (HPT) axes and disturbances in serotonergic neurotransmission in suicidal patients. In order to investigate the biological correlates of suicidal ideation, we measured the following: basal thyrotropin-secreting hormone (TSH), free thyroxine (FT4), pre- and postdexamethasone cortisol, adrenocorticotropic hormone (ACTH) levels, the circulating concentrations of total L-tryptophan (L-TRP) and the ratio between L-TRP and competing amino acids (CAA). The subjects were 17 suicidal and 17 nonsuicidal major depressed females matched for age and severity of illness. We found no significant differences in any of the above-mentioned biological data between patients with suicidal ideation and those without. PMID- 2548372 TI - Effect of acute ether or restraint stress on plasma corticotropin-releasing hormone, vasopressin and oxytocin levels in the rat. AB - Ether and restraint stress-induced peripheral plasma corticotropin releasing hormone (CRH), arginine vasopressin (AVP), oxytocin (OXY) and adrenocorticotropin (ACTH) levels were measured by radioimmunoassays. Plasma CRH, AVP, OXY and ACTH rose to approximately twice the level of control rats 2 min after the onset of a 1-min exposure to ether. Plasma CRH rose further 5 min after the onset of ether stress, while plasma AVP and OXY returned to the baseline levels at 5 min. Plasma CRH, OXY and ACTH showed significant elevation 2 min after the onset of restraint stress, while plasma AVP did not show a significant change. Plasma OXY and ACTH rose further 5 min after the onset of restraint stress, whereas plasma CRH returned to baseline levels. CRH and OXY concentrations in the hypothalamic median eminence decreased 5 min after the onset of ether exposure and restraint, while the AVP concentration did not differ from control levels. The results, including the discrepancy between plasma CRH and ACTH 5 min after stress, suggest that CRH in the peripheral plasma is derived from both hypothalamic and extrahypothalamic tissues. The levels of stress-induced CRH in the peripheral plasma were sufficient to stimulate ACTH release. These results suggest that ether and restraint stress elevate plasma CRH shortly after the onset of the stress, and that this elevation in the plasma CRH level is at least partly responsible for stress-induced ACTH secretion. PMID- 2548373 TI - Elevated IgA antibodies to Epstein-Barr virus in children with chronic active Epstein-Barr virus infection. AB - Anti-Epstein-Barr virus (EBV) antibodies were tested in 11 children with chronic active EBV infection. Anti-virus capsid antigen (VCA)-IgG antibody titers ranged from 1:640 to 1:10,240. Anti-VCA-IgM antibody was consistently positive in 5 of the 11 patients; anti-VCA-IgA antibody was consistently positive in 6 of the 10 patients; anti-early antigen (EA)-IgG antibody was consistently positive in 10 of the 11 patients and anti-EA-IgA antibody was consistently positive in 4 out of the 7 patients. Anti-EBV nuclear antigen (EBNA) antibody was not detected in two patients. Consistently positive anti-VCA-IgA- and anti-EA-IgA- antibody may be a characteristic feature of abnormal antibody responses in severe chronic active EBV-infection in childhood. PMID- 2548374 TI - Dog and rat kidney proximal tubule cells in culture: responses to parathyroid hormone. AB - We have described and compared culture systems for proximal tubule cell (PTC) preparations from dog and rat kidney. Cells were prepared from kidney cortex by enzyme digestion and purified on Percoll density gradient. The dog PTC and rat PTC differed in their growth characteristics in culture. Although the dog PTC tended to overgrow the contaminating fibroblastic cells, the rat PTC tended to be overgrown by the latter cells when cultured in medium containing 15% fetal calf serum (FCS). Cultures of rat PTC in serum-free medium or medium containing only 2% FCS yielded only epithelium-like cells exhibiting characteristics of cells that are proximal tubular in origin. These properties include protrusion of microvilli, high alkaline phosphatase activity, ability to transport sugar, and responsiveness to parathyroid hormone (PTH) in terms of cAMP production. The time course and dose-response curves of PTH-stimulated cAMP accumulation were studied in dog and rat PTC. The estimated half-maximal concentrations (Kact) for PTH in dog and rat PTC were 1.2 and 100 nM, respectively. Both values are within the range reported in the literature for the respective renal membrane preparations. In addition to our previously reported data on dog PTC, this study revealed the presence of PTH-inhibitable 25-hydroxyvitamin D3-24-hydroxylase in dog PTC. These two model cell culture systems should prove useful in studying PTH action in renal cells. PMID- 2548375 TI - Accelerated bone resorption in senescence-accelerated mouse (SAM-P/6). AB - Age-associated changes in the femoral bone and in urine and serum composition were studied to understand the low bone mass in a substrain of senescence accelerated mouse (SAM), SAM-P/6. Age-matched normal SAM-R/1 mice were used as controls. After 13 weeks of age, the concentration of hydroxyproline in the femur of SAM-P/6 was slightly but significantly lower. The urinary excretion rates of cAMP and calcium were higher in SAM-P/6 throughout the whole experimental period, and those of hydroxyproline and phosphorus were higher after 8 weeks. The serum concentrations of calcium and inorganic phosphorus of SAM-P/6 were higher at 13 and 20 weeks. At 5 weeks, the serum alkaline phosphatase and tartrate-resistant acid phosphatase (TRAP) activities of SAM-P/6 were significantly higher. Furthermore, the serum of SAM-P/6 significantly stimulated calcium release from cultured fetal rat ulna. Since urinary cAMP and TRAP in the serum reflect the circulating level of parathyroid hormone and osteoclastic function, respectively, the present results suggest that, in SAM-P/6, accelerated bone resorption produced by a putative hyperparathyroid state causes the decrease in bone mass. PMID- 2548376 TI - Paragangliomas of the jugular bulb and carotid body: MR imaging with short sequences and Gd-DTPA enhancement. AB - Twenty-six patients with glomus jugulare (16), glomus tympanicum (three), or carotid glomus (seven) tumors were examined with contrast-enhanced CT scans and MR scans without and with Gd-DTPA. MR and CT scans had similar sensitivities, but the enhanced MR scans were diagnostically more specific than either CT or nonenhanced MR. Dynamic MR scanning permitted measurement of the degree of Gd DTPA enhancement over time. We recommend contrast-enhanced MR with short sequences and a dynamic approach in patients with suspected carotid, tympanic, and jugular paragangliomas. PMID- 2548377 TI - Calcification of metastatic mucinous carcinoma of the liver after embolization. PMID- 2548378 TI - Transcatheter arterial embolization for the treatment of ruptured hepatocellular carcinoma. PMID- 2548379 TI - Intraluminal irradiation for inoperable obstructing endobronchial carcinoma of the lung. AB - Between April 1, 1987 and August 31, 1988, 31 patients with inoperable symptomatic obstructing carcinoma of the lung underwent 79 intraluminal irradiation sessions in an effort to re-establish patency of the airway. Palliation was excellent, 74% of the patients had complete relief of atelectasis, while two patients with extensive tracheal disease experienced little objective response. There was little morbidity, other than that associated with bronchoscopy. Overall, survival at 6 months was 55% with 22% of patients at risk for 18 months surviving. This compares favorably with the more complicated technique of laser resection for relieving malignant airway obstruction. PMID- 2548380 TI - Structure and function of alpha-adrenergic receptors. AB - Alpha 1- and alpha 2-adrenergic receptors are the initial recognition sites on a wide variety of catecholamine-responsive target cells. This article addresses several major questions related to subtypes, structure, signal transduction mechanisms, and regulation of alpha 1- and alpha 2-adrenergic receptors. The application of biochemical and cell and molecular biologic techniques has provided many new insights regarding alpha-adrenergic receptors. Two (and perhaps three) distinct alpha 2-adrenergic receptor subtypes have been identified, and subtypes may exist for alpha 1-adrenergic receptors as well. These multiple subtypes imply much greater diversity among alpha-adrenergic receptors than among beta-adrenergic receptors. Alpha-adrenergic receptors are membrane glycoproteins with several common structural features (including seven membrane-spanning domains with extracellular amino terminus and intracellular carboxyl terminus) that are shared with other types of membrane receptors linked to guanine nucleotide-binding regulatory (G) proteins. These G proteins appear to link alpha adrenergic receptors to multiple effector systems, including enzymes such as adenylate cyclase and phospholipases, and ion channels. The receptors themselves are dynamic entities, the number of which is regulated as a consequence of a poorly understood life cycle. Although unproven, it seems likely that several important clinical disorders represent alterations in alpha-adrenergic receptors themselves or in the G proteins or effector systems to which these receptors couple. New tools for studying receptor structure and function should help clarify the numerous, inadequately understood issues regarding alpha-adrenergic receptors and their possible alteration in disease states. PMID- 2548381 TI - Mechanisms contributing to the arrhythmogenic influences of alpha 1-adrenergic stimulation in the ischemic heart. AB - The majority of deaths associated with ischemic heart disease occur suddenly because of disturbances in cardiac rhythm culminating in ventricular fibrillation. Past research has focused on elucidating the biochemical membrane mechanisms responsible for the adverse electrophysiologic alterations in the ischemic heart, with major emphasis on the influence of adrenergic neural factors. It has been demonstrated that both alpha 1-and beta-adrenergic mechanisms contribute to arrhythmogenesis in the ischemic heart. In the normal heart, alpha 1-adrenergic input has very little effect on electrophysiologic indices. However, during early ischemia and reperfusion, enhanced alpha 1 adrenergic responsivity associated with a twofold reversible increase in alpha 1 adrenergic receptors in vivo has been demonstrated. Likewise, in a variety of species, alpha 1-adrenergic inhibition with prazosin markedly decreases the incidence of malignant ventricular arrhythmias associated with either myocardial ischemia or subsequent reperfusion. One major manifestation of alpha 1-adrenergic receptor activation during reperfusion of ischemic myocardium is an increase in intracellular calcium ion (Ca2+). It has been demonstrated that reperfusion of ischemic myocardium increases intracellular Ca2+ in reversibly injured tissue, and that the gain in intracellular Ca2+ is prevented by alpha 1-adrenergic inhibition with hydroxyphenylethyl aminomethyl tetralone, even when administered just prior to reperfusion. Subsequently, it was demonstrated that the alpha 1 adrenergic-induced increase in mitochondrial Ca2+ contributes to the decline in mitochondrial function. These findings suggest that even single-dose intervention with alpha 1-adrenergic inhibitors may improve markedly the functional recovery and extent of ultimate necrosis in humans after coronary thrombolysis. To investigate the mechanisms responsible for the increase in alpha 1-adrenergic receptors during ischemia, we used isolated adult canine ventricular myocytes exposed to hypoxia. Thirty minutes of hypoxia at 25 degrees C or 10 minutes of hypoxia at 37 degrees C resulted in a threefold reversible increase in the density of surface alpha 1-adrenergic receptors and a threefold increase in the cellular content of long-chain acylcarnitines. Inhibition of carnitine acyltransferase I abolished not only the accumulation of long-chain acylcarnitines during hypoxia but also the increase in alpha 1-adrenergic receptors. Exposure of normoxic myocytes to exogenous long-chain acylcarnitines (1 mumol/liter) for 10 minutes also increased alpha 1-adrenergic receptor number. These findings indicate that the sarcolemmal accumulation of long-cha PMID- 2548382 TI - Phase II trial of ifosfamide and mesna in mixed mesodermal tumors of the uterus (a Gynecologic Oncology Group study). AB - A phase II trial of ifosfamide (isophosphamide, NSC 109724) and mesna (2 mercaptoethane sodium sulfonate, NSC 113891) in women with advanced or recurrent mixed mullerian tumors of the uterus was conducted by the Gynecologic Oncology Group. The starting dose of ifosfamide was 1.5 gm/m2 daily, intravenously, for 5 days. The starting dose of ifosfamide was reduced 1.2 gm/m2 daily in patients who had received prior radiotherapy. Mesna was given intravenously immediately and at 4 and 8 hours after the administration of ifosfamide. Each mesna dose was 20% of the total daily dose of ifosfamide. Twenty-nine patients are evaluable for toxicity, and 28 patients are evaluable for response. Twenty-one patients had received prior abdominal hysterectomy, and eight patients had prior radiotherapy. Thirteen tumors were homologous and 15 heterologous. Gynecologic Oncology Group grade 3 or 4 granulocytopenia occurred in seven (25%) patients and two (7.1%) had grade 3 or 4 thrombocytopenia. Two patients (7.1%) had grade 3 or 4 neurotoxicity. One patient experienced lethargy and confusion that responded to discontinuation of the ifosfamide. A second patient developed progressive cerebellar dysfunction, left hemiparesis, and coma. This patient died after 3 days of therapy. Complete responses were seen in five (17.9%) patients and partial responses occurred in four (14.3%) patients for a total response rate of 32.2%. These results indicate that ifosfamide is an unusually active drug in patients with advanced or recurrent mixed mullerian tumors of the uterus. Studies with combination regimens incorporating ifosfamide are warranted. The toxicity of ifosfamide in Gynecologic Oncology Group studies is being evaluated retrospectively. PMID- 2548383 TI - Erythrocyte cation metabolism in preeclampsia. AB - To determine if there are abnormalities in cellular cation regulation in pregnancy-induced hypertension, erythrocyte intracellular levels of calcium, magnesium, sodium, and potassium and circulating parathyroid hormone and "endoxin" were examined in 13 women with pregnancy-induced hypertension and 34 control subjects matched for gestational age (greater than or equal to 35 weeks). Both endoxin and parathyroid hormone levels were higher in patients with pregnancy-induced hypertension than in control subjects (endoxin, 294 +/- 34 vs. 210 +/- 19 pg/ml, p less than 0.05; parathyroid hormone, 0.65 +/- 0.05 vs. 0.60 +/- 0.03 ng/ml); the increase was significant only for endoxin. Intracellular calcium was higher in the patients with pregnancy-induced hypertension (0.033 +/- 0.010 vs. 0.015 +/- 0.001 mEq/L, p less than 0.05, in the patients with pregnancy induced hypertension and control patients, respectively) but intracellular sodium, potassium, and magnesium levels were not different. This intracellular calcium elevation may be caused directly by the increase in parathyroid hormone or indirectly by the observed elevation in endoxin. Our data indicate that the observed effect is specific because no changes in intracellular sodium, potassium, or magnesium levels were found. PMID- 2548384 TI - Atrial natriuretic peptide receptors in guinea pig placentas. AB - The multiple physiologic effects of atrial natriuretic peptide are mediated through specific atrial natriuretic peptide plasma membrane receptors. We have attempted to determine whether atrial natriuretic peptide receptors are present in guinea pig placentas and subplacentas at approximately 45 and 65 days' gestation. A microsomal plasma fraction was prepared from each component. Atrial natriuretic peptide receptors were detected by in vitro radioligand techniques with alpha-human atrial natriuretic peptide labeled with iodine 125. In placentas we identified one set of specific binding sites for atrial natriuretic peptide at both gestational ages. Although the maximal concentration of the receptors did not change with advancing gestation, the dissociation constant was higher at 65 than at 45 days' gestation. In subplacentas two sets of binding sites were identified, one of low capacity-high affinity and the other of low affinity-high capacity. The maximal concentration of the atrial natriuretic peptide receptors did not change as a function of gestation. However, the dissociation constant, for both the high- and low-affinity sites, significantly decreased with advancing gestation. PMID- 2548385 TI - Na+-H+ exchange in isolated myocytes from adult rat heart. AB - The intracellular pH (pHi) in ventricular myocytes isolated from adult rat heart was measured in suspension using the pH-sensitive dye 2',7'-bis(2-carboxyethyl) 5(6)-carboxyfluorescein (BCECF). Steady-state pHi in bicarbonate-free media [extracellular pH (pHo) = 7.4] was 7.16 +/- 0.11 at 37 degrees C. With the use of the ammonium chloride prepulse technique, pHi was acidified, and the rate of return to resting pHi was determined. Initial rate analysis of the recovery was used to characterize the kinetics of proton net efflux via the Na+-H+ exchanger. At pHo = 7.4, proton extrusion was stimulated by extracellular sodium with a K1/2 = 58 +/- 16 mM and a maximal rate of recovery of 55 +/- 7 mmol/(1 cell H2O.min). Amiloride, which inhibited greater than 90% of the observed proton movements, was a competitive inhibitor with respect to Nao, with an inhibition constant of 3.5 microM. Proton net efflux was also inhibited by extracellular protons, with a maximal flux occurring above pHo = 8 and no net efflux occurring below pHo = 6.0. Efflux was stimulated by intracellular protons over a much narrower range (pHi = 6.6-7.1). This steep dependence indicates the involvement of at least one additional proton binding site besides the intracellular transport site, in accordance with the kinetic behavior observed in other cell systems. PMID- 2548386 TI - Leukotriene D4-induced proliferation of glomerular epithelial cells: PKC- and Na+ H+ exchanger-mediated response. AB - The growth-promoting effect of leukotriene D4 (LTD4) has been observed in a variety of cells, including human glomerular epithelial cells. The purpose of this study was to determine the mechanisms underlying this process. LTD4 induction of [3H]thymidine uptake in human glomerular epithelial cells was blocked by the LTD4 receptor antagonist L648,051 when added in a 50-fold excess and by pertussis toxin. Neither drug affected basal DNA synthesis. These results suggest that the LTD4-mediated signal transduction implies activation of a GTP binding protein that is coupled to a specific receptor. The possible role of protein kinase C (PKC) activation was also studied. In the presence of the PKC inhibitor H-7 or after downregulation of PKC levels by chronic treatment with phorbol ester, stimulation of [3H]thymidine uptake by LTD4 was greatly inhibited. Moreover, treatment of the cells by LTD4 resulted in a time-dependent increase of cytosolic PKC activity, whereas addition of phorbol 12-myristate 13-acetate reduced this activity. Therefore PKC-dependent mechanisms are likely to mediate the growth-promoting effect of LTD4. Finally, three approaches were used to determine the potential role of the Na+-H+ exchanger. First, progressive removal of extracellular Na+ using N-methyl-D-glucamine+ as a substitute inhibited LTD4 induced [3H]thymidine uptake with a 50% inhibitory concentration (IC50) of 85 mM. Second, addition of amiloride reduced the LTD4 growth effect with an IC50 of 6.5 microM, whereas three amiloride analogues exhibited lower IC50 values in accordance with their greater affinity for the Na+-H+ exchanger.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548387 TI - Conductive pathways for HCO3- in basolateral membrane of salamander intestinal cells. AB - To understand the route of HCO3- exit across the basolateral membrane of Amphiuma small intestinal cells during active H+ secretion the influence of bath HCO3- on serosal membrane potential (Vs) was measured using conventional microelectrodes. The villus sheet preparation was used, which allowed direct access to the basal membrane for microelectrode impalement. When tissues were incubated in Cl(-)-free (SO2-4 based) medium, Vs averaged -85.8 mV. Elevation of serosal bath [HCO3-] at constant CO2 increased Vs; reduction of the [HCO3-] reduced Vs and increased the fractional resistance of the serosal membrane. When serosal bath HCO3- and CO2 were completely replaced, Vs was -44.8 mV. The serosal membrane was also depolarized on complete replacement of medium HCO3- and CO2 when bath pH was buffered with phosphate. 4,4'-Dinitro-2,2'-stilbene disulfonate (DNDS) and acetazolamide blocked the decline in Vs produced by lowering serosal medium [HCO3 ]. Replacement of serosal Na+ reduced Vs 32.7 mV. DNDS blocked this response. It is concluded that a Na+-dependent, stilbene-sensitive HCO3- exit pathway resides in the basolateral membrane of the intestinal cells. Possibly there is also a parallel, conductive pathway for HCO3-. PMID- 2548388 TI - Excess membrane cholesterol alters calcium channels in arterial smooth muscle. AB - We studied the effects of cholesterol enrichment on arterial function by evaluating its effects on 45Ca2+ uptake and tension development in the carotid artery of the rabbit. Arterial segments were enriched with cholesterol in vitro, using media containing liposomes composed of free (unesterified) cholesterol (FC) and phospholipid (PL) in a 2:1 molar ratio. Control segments were simultaneously perfused with 0.5:1 liposomal medium to compare the possible effects of PL. Rings from these arteries were then tested for basal and activated Ca2+ uptake and for contractile responses to norepinephrine (NE) and KCl. We found elevated 45Ca2+ uptake under basal and NE-activated conditions along with an increased contractile sensitivity (4-fold) to NE. These alterations correlated with a 78% increase in the FC/PL ratio reflecting cholesterol enrichment of cellular membranes. Cholesterol enrichment did not alter resting or maximal tensions, K+ activated Ca2+ uptake, or contractile sensitivity to K+. Pretreatment with 1 microM diltiazem abolished the cholesterol-induced increase in basal as well as NE-activated 45Ca2+ uptake but had no effect on either uptake in control vessels. These studies suggest that excess membrane cholesterol selectively increases NE contractile sensitivity by increasing basal or NE-activated Ca2+ influx (or both) as a result of fundamental alteration in the calcium channels in arterial smooth muscle cell membrane. PMID- 2548389 TI - Continuous membrane voltage recordings in A10 vascular smooth muscle cells: effect of AVP. AB - Continuous membrane voltage (V) recordings were obtained in A10 vascular smooth muscle cells (rat aorta) using glass microelectrodes. Resting membrane voltage in 262 impalements averaged 54.0 +/- 0.4 (SE) mV. Relative K+ conductance was characterized, and the contribution of electrogenic Na+-K+-ATPase to membrane voltage was investigated. Action potentials could be induced by application of 1 mM barium or 10(-4) M acetylcholine. In a few recordings, spontaneous spike activity occurred, and this could be abolished by 5 mM MgCl2 or by removal of extracellular Ca2+. Barium-induced action potentials were not dependent on the presence of extracellular Na+ and not inhibitable by 10(-6) M tetrodotoxin. Application of 10(-6) M [Arg8] vasopressin (AVP) for 30 s caused a typical biphasic membrane voltage response with an initial transient hyperpolarization of -9.5 +/- 1.1 mV and a more sustained subsequent depolarizing response averaging 28.2 +/- 1.3 mV (mean +/- SE, n = 58). The effect of AVP on membrane voltage was blocked by the V1-antagonist [beta-mercapto-beta,beta cyclopentamethylenepropionyl1,O-Me- Tyr2,Arg8]vasopressin. The initial hyperpolarizing component of the membrane voltage response to AVP became more prominent when V was predepolarized, for example, by a preceding AVP application. However, when AVP was applied during high K+ depolarization or in the presence of quinidine (1 mM), the initial hyperpolarizing response was practically abolished. The time course of the initial hyperpolarization was shown to be similar to the calcium transient observed in fura-2-loaded A10 cell suspensions after the application of AVP. We conclude that the initial AVP-induced hyperpolarization in A10 cells corresponds to an activation of Ca2+-activated K+ channels. PMID- 2548390 TI - Contractile force and intracellular Ca2+ during relaxation of canine tracheal smooth muscle. AB - Muscle strips loaded with the Ca2+ indicator aequorin were studied in vitro to determine the effects of inhibitory stimuli on force and cytosolic free Ca2+. In muscles contracted isometrically with acetylcholine (ACh), 5-hydroxytryptamine (5 HT), carbachol, decreases in muscle force caused by isoproterenol (10(-5) M) or forskolin (10(-5) M) were accompanied by proportional decreases in aequorin luminescence. A similar relationship between decreases in muscle force and aequorin luminescence was observed when muscles were relaxed by stimulating Na+ K+-ATPase activity. These results suggest that the Ca2+ sensitivity of contractile proteins was not decreased during adenosine 3',5'-cyclic monophosphate (cAMP)-dependent relaxation. However, aequorin luminescence did not decrease when muscles contracted by K+ depolarization were relaxed with isoproterenol. Incubation of muscles in forskolin depressed increases in both force and aequorin luminescence in response to 5-HT or ACh. Incubation of muscles in isoproterenol had a similar effect on responses to 5-HT but depressed increases in force without depressing increases in luminescence in response to ACh. Results indicate that under most conditions the reduction of cytosolic Ca2+ plays an important role in the cAMP-dependent relaxation of canine tracheal smooth muscle. PMID- 2548391 TI - Propionate induces cell swelling and K+ accumulation in shark rectal gland. AB - Small organic anions have been reported to induce cell solute accumulation and swelling. To investigate the mechanism of swelling, we utilized preparations of rectal gland cells from Squalus acanthias incubated in medium containing propionate. Propionate causes cells to swell by diffusing across membranes in its nonionic form, acidifying cell contents, and activating the Na+-H+ antiporter. The Na+-H+ exchange process tends to correct intracellular pH (pHi), and thus it maintains a favorable gradient for propionic acid diffusion and allows propionate to accumulate. Activation of the Na+-H+ antiport also facilitates Na+ entry into the cell and Nai accumulation. At the same time Na+-K+-ATPase activity, unaffected by propionate, replaces Nai with Ki, whereas the K+ leak rate, decreased by propionate, allows Ki to accumulate. As judged by 86Rb+ efflux, the reduction in K+ leak was not due to propionate-induced cell acidification or reduction in Cli concentration. Despite inducing cell swelling, propionate did not disrupt cell structural elements and F actin distribution along cell membranes. PMID- 2548392 TI - Fast Na+ and slow Ca2+ channels in single uterine muscle cells from pregnant rats. AB - Whole cell voltage-clamp method was applied to single smooth muscle cells freshly isolated from the longitudinal layer of 18-day pregnant rat uterus. Inward currents were isolated after outward currents were minimized by use of high Cs+ in the pipette solution and 4-aminopyridine (3 mM) in the bath solution. Depolarizing pulses, applied from a holding potential of -90 mV, evoked two types of inward current, fast and slow. The fast inward current decayed and disappeared within 30 ms and depended on extracellular Na+ concentration. This fast current was inhibited by tetrodotoxin (TTX) dose dependently (KD = 27 nM). These results suggest that the fast inward current was a TTX-sensitive Na+ channel current. In contrast, the slow inward current decayed slowly, dependent on extracellular Ca2+ (or Ba2+) concentration, and was inhibited by the Ca2+ channel blocker, nifedipine, dose dependently (10 nM-10 microM). These results suggest that the slow inward current was a Ca2+ channel current. A fast-inactivating Ca2+ channel current was not evident when Ba2+ was the charge carrier. We conclude that the major ion channels in the cell membrane of pregnant rat uterus, which generate inward currents, are TTX-sensitive fast Na+ channels and dihydropyridine sensitive slow Ca2+ channels (L-type, high-threshold type). PMID- 2548393 TI - Stretch-related changes in lung cAMP after partial pneumonectomy. AB - In rats, left pneumonectomy (PNX) initiates rapid compensatory hyperplastic growth of the right lung. Previous work indicated that increased polyamine uptake associated with post-PNX distortion or "stretch" of the remaining tissue and altered adenosine 3',5'-cyclic monophosphate (cAMP) metabolism may play a role in the early phase of the growth response. Evidence was sought to link these observations with the goal of understanding control of compensatory growth. At day 1 or 3 after left PNX in vivo, increased right lung cAMP was associated with activation of the cAMP-dependent protein kinase (PKa). Total PKa activity was unaffected, but the PKa activity ratio (-cAMP/+cAMP) doubled, providing evidence for PNX-associated activation of the kinase in vivo. Neither cAMP nor PKa was altered in sham-operated control lungs. To determine whether increased distension of the right lung after PNX might initiate these changes, lungs of unoperated animals were perfused 40 min in vitro, either without ventilation or with a distending constant positive airway pressure (CPAP) of 20 cmH2O. CPAP rapidly increased uptake of the polyamine spermidine (SPD; 1.5 microM), tissue cAMP, and the PKa activity ratio, in a manner similar to that observed after PNX. Both tissue cAMP and the kinase activity ratio increased with 1 microM forskolin (FSK), as expected, but SPD uptake was unaffected by FSK. FSK did not diminish the CPAP-associated elevations of SPD uptake (P less than 0.01) or cAMP (P less than 0.05), but the kinase was not further activated by CPAP with FSK present.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548394 TI - New form of pseudohypoparathyroidism with abnormal catalytic adenylate cyclase. AB - Patients with pseudohypoparathyroidism type Ia have resistance to multiple hormones because of deficient activity of the stimulatory guanine nucleotide binding protein (Gs) that couples membrane receptors to activation of adenylate cyclase. However, in a subset of patients with pseudohypoparathyroidism who have resistance to multiple hormones yet possess normal erythrocyte membrane Gs activity, the biochemical abnormality responsible for hormone resistance has remained undefined. Cultured skin fibroblasts were derived from a patient with this atypical form of pseudohypoparathyroidism. In the patient's fibroblast membranes, adenylate cyclase stimulation mediated by Gs after fluoride ion treatment produced only 52% of normal activity, yet fibroblast membrane Gs activity measured by cyc- complementation was normal. Activation of the catalytic unit of adenylate cyclase with manganese produced 49% of normal activity; manganese plus forskolin produced 54% of normal adenylate cyclase activity. beta Adrenergic receptor coupling to Gs and phosphodiesterase activity were normal. A defect in the catalytic unit of adenylate cyclase can account for these results and may be a mechanism for clinical resistance to multiple hormones that act through adenylate cyclase. PMID- 2548395 TI - Na+-K+-ATPase pump function in rat brain synaptosomes is different in males and females. AB - To understand the increased morbidity and mortality associated with acute hyponatremia in young women, we characterized the Na+-K+-adenosinetriphosphtase (ATPase) pump in rat brain synaptosomes to determine if this adaptive mechanism was different between the sexes. Veratridine-stimulated sodium (Na+) uptake was significantly greater (P less than 0.001) in females than in males (8.08 +/- 0.3 vs. 5.56 +/- 0.4 nmol/mg protein), suggesting either an increased rate of Na+ uptake and/or decreased extrusion of Na+ by the Na+-K+-ATPase pump in females. Uptake rate was determined by measuring Na+ transport at 5 s, and it was found to be twice as large in females as in males (1.01 +/- 0.2 vs. 0.46 +/- 0.1 nmol/mg protein). However, in the presence of 2.5 mM ouabain, uptake in both groups were similar, suggesting that the difference was probably due to decreased function of the Na+-K+-ATPase pump in females. Transport evaluation of the Na+-K+-ATPase pump showed ouabain-sensitive K+ uptake in males to be significantly greater (P less than 0.001) than in females (10.53 vs. 4.97 nmol/mg protein), and ouabain sensitive Na+ uptake in inverted synaptosomes was 70% greater in males (4.00 vs. 2.37 nmol/mg protein). [3H]ouabain binding studies showed maximum binding capacity in males and females to be similar (103 +/- 12 vs. 110 +/- 15 pmol/mg protein), whereas the dissociation constant was significantly (P less than 0.005) greater in males (109 +/- 8 vs. 82 +/- 6 nM).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548396 TI - Alpha-adrenergic regulation of Na-Cl cotransport in human airway epithelium. AB - The demonstration of abnormal beta-adrenergic and cAMP-modulated apical Cl- channels in cystic fibrosis (CF) airway epithelial cells suggests that other transporters, which are required for Cl- secretion, may also be abnormally regulated. A basolateral cotransporter was investigated by determining the initial rate of 36Cl efflux from cells isolated from CF nasal polyps or trachea and non-CF trachea. Cells were preequilibrated with radioactive tracer at 25 degrees C, and tracer transport was initiated by 10-fold dilution of an aliquot of cells in radioisotope-free medium. The initial rate of Cl transport was calculated from the linear portion of the efflux curves. In CF and non-CF cells, base-line Cl- transport was not blocked by furosemide but was stimulated twofold by l-epinephrine in Ca2+-deficient and Ca2+-replete transport medium. In both types of cells, furosemide blocked 70 and 77%, respectively, of the stimulated Cl transport. Prazosin, an alpha 1-adrenergic antagonist, blocked the effects of l epinephrine and methoxamine, an alpha 1-adrenergic agonist, stimulated prazosin- and furosemide-sensitive Cl transport. Ionomycin mimicked the effects of l epinephrine. l-Isoproterenol, a beta-adrenergic agonist, did not affect Cl transport. The results of this study indicate an alpha 1-adrenergic stimulation of furosemide-sensitive Cl transport in human airway epithelium that functions normally in CF airway epithelial cells. The transport mechanism is probably a Na Cl or Na-K-2Cl cotransport located in the basolateral membrane and requires elevated intracellular Ca2+ for activation. PMID- 2548397 TI - Role of organic anions in renal response to dietary acid and base loads. AB - Bicarbonate is formed when organic anions are oxidized systemically. Therefore, changes in organic anion excretion can affect systemic acid-base balance. To assess the role of organic anions in urinary acid-base excretion, we measured urinary excretion in control rats, NaHCO3-loaded rats, and NH4Cl-loaded rats. Total organic anions were measured by the titration method of Van Slyke. As expected, NaHCO3 loading increased urine pH and decreased net acid excretion (NH4+ + titratable acid - HCO3-), whereas NH4Cl loading had the opposite effect. Organic anion excretion was increased in response to NaHCO3 loading and decreased in response to NH4Cl loading. We quantified the overall effect of organic ion plus inorganic buffer ion excretion on acid-base balance. The amounts of organic anions excreted by all animals in this study were greater than the amounts of NH4+, HCO3-, or titratable acidity excreted. In addition, in response to acid and alkali loading, changes in urinary organic anion excretion were 40-50% as large as changes in net acid excretion. We conclude that, in rats, regulation of organic anion excretion can contribute importantly to the overall renal response to acid-base disturbances. PMID- 2548398 TI - Sodium transfer from endolymph through a luminal amiloride-sensitive channel. AB - An in vitro preparation of frog semicircular canal was devised to study the mechanisms of Na transport across the labyrinthine epithelium. When the lumen of the semicircular canal was filled with perilymph-like solution, the structure was able to secrete K into and to absorb Na from the lumen and to generate a lumen positive transepithelial potential. When the lumen of the semicircular canal was filled with endolymph-like solution, the electrochemical composition of the luminal fluid was partly maintained up to 2 h. In this last experimental condition net and unidirectional fluxes were calculated in absence or presence of transport inhibitors, separately for the ampulla and for the nonampullar part of the canal. Amiloride (10(-5) M) but not dimethyl amiloride (10(-5) M) inhibited 60% of the unidirectional Na efflux out of endolymph; this Na efflux decrease resulted in an increase of the inward net Na flux. The net Na flux was also increased after abluminal application of ouabain (10(-3) M), furosemide (10(-4) M), and bumetanide (10(-6) M). This study validates this isolated preparation as a suitable tool for the study of endolymph secretion, confirms that the secretion of endolymph is achieved in the ampulla, and provides evidence for an apical amiloride-sensitive Na channel through which Na is transferred out of endolymph along an electrochemical gradient provided by the activity of the abluminal Na+ K+-ATPase. PMID- 2548399 TI - Effects of beta 1-adrenergic blockade on glomerular dynamics and angiotensin II response. AB - Adrenergic activity regulates renal function by several mechanisms. Renal nerves not only exert vasoconstrictor functions but also may influence glomerular hemodynamics by beta-adrenergic activity, especially via the effects on renin angiotensin activity. Little is known of the specific glomerular hemodynamic alterations resulting from beta 1-adrenergic blockade. Current studies examined the effects of 4-6 days of treatment with atenolol (50 mg/kg), a beta 1-selective adrenergic antagonist, on glomerular hemodynamics in plasma volume-expanded Munich-Wistar rats. Atenolol treatment reduced blood pressure both in the awake state and during micropuncture. This reduction in blood pressure contributed to a decrease in nephron filtration rate (48 +/- 1 in untreated rats vs. 40 +/- 1 nl.min-1.g kidney wt-1 in the atenolol-treated group, P less than 0.05) by reduction in nephron plasma flow (182 +/- 2 vs. 154 +/- 4 nl.min-1.g kidney wt-1 in the atenolol-treated rats). No other determinant of glomerular ultrafiltration was influenced by atenolol treatment. Since beta 1-adrenergic blockade may influence the generation of angiotensin II, the response to angiotensin II infusion was assessed and found not to differ from control untreated animals. These studies demonstrate that beta 1-receptor blockade reduced nephron filtration rate by decreasing mean arterial blood pressure and nephron plasma flow without significant modifications in vascular resistance and the glomerular hydrostatic pressure gradient. PMID- 2548400 TI - Dual regulation of insulin-like growth factor I expression during renal hypertrophy. AB - We examined the regulation of insulin-like growth factor I (IGF-I) in kidney during the renal hypertrophy produced by two different experimental models: growth hormone treatment of hypophysectomized rats and compensatory hypertrophy subsequent to unilateral nephrectomy. Immunostaining for IGF-I in collecting ducts was enhanced in kidneys from growth hormone-repleted hypophysectomized rats, and the levels of IGF-I mRNAs were increased. In compensatory hypertrophy, no enhancement of the intensity of immunostaining was observed in kidneys of nephrectomized rats until 5 days postnephrectomy, at which time immunostainable IGF-I was increased markedly in medullary collecting ducts of hypertrophied kidneys compared with kidneys from sham-operated animals. No difference in steady state levels of any IGF-I mRNA species was detected in whole kidneys or in collecting ducts from nephrectomized or sham-operated rats at any time postnephrectomy. Our findings demonstrate an increase in both IGF-I mRNA and in immunostainable IGF-I in collecting duct in the setting of growth hormone-induced renal hypertrophy but suggest that other, possibly translational, mechanisms underlie the induction of IGF-I synthesis during compensatory hypertrophy. PMID- 2548401 TI - Renal reabsorption of phosphate during development: transport kinetics in BBMV. AB - The mechanism responsible for enhanced reabsorption of phosphate (Pi) in growing animals was assessed in renal brush-border membrane vesicles (BBMV) prepared from 3- to 14-day-old and greater than 57-day-old guinea pigs. On standard diet, Vmax (pmol.mg-1.s-1) of Na+-Pi cotransport was higher (P less than 0.001) in newborn (650 +/- 77) than in adult (144 +/- 17) but Vmax of Na+-glucose cotransport did not differ with age. Low dietary Pi did not affect significantly Vmax of Na+-Pi cotransport in the newborn (P greater than 0.8) but increased it in the adult (to 318 +/- 32, P less than 0.05). A high Pi intake resulted in a smaller relative decrease in Vmax in the newborn than in the adult (27 vs. 44%, P less than 0.05). In the newborn, the serum Pi (mM) decreased on a low-Pi diet (from 1.8 +/- 0.1 to 0.8 +/- 0.1, P less than 0.001) and rose by twofold on the high-Pi diet (to 3.5 +/- 0.2, P less than 0.001). In the adult, there were no significant changes in serum Pi with changes in Pi intake (P greater than 0.5). Thus in the newborn the Na+-Pi cotransport system is characterized by high transport capacity but low adaptability to changes in dietary Pi. PMID- 2548403 TI - Autoradiographic demonstration of oxytocin-binding sites in the macula densa. AB - Specific oxytocin (OT)-binding sites were localized in the rat kidney with use of a selective 125I-labeled OT antagonist (125I-OTA). High concentrations of OT binding sites were detected on the juxtaglomerular apparatus with use of the conventional film autoradiographic technique. No labeling occurred on other renal structures. The cellular localization of the OT binding sites within the juxtaglomerular apparatus was studied in light microscope autoradiography, on semithin sections from paraformaldehyde-fixed kidney slices incubated in the presence of 125I-OTA. These preparations revealed selective labeling of the macula densa, mainly concentrated at the basal pole of the cells. Control experiments showed first that 125I-OTA binding characteristics were not noticeably altered by prior paraformaldehyde fixation of the kidneys and second that autoradiographic detection of the binding sites was not impaired by histological treatments following binding procedures. In view of the role of the macula densa in the tubuloglomerular feedback, the putative OT receptors of this structure might mediate the stimulatory effect of OT on glomerular filtration. PMID- 2548402 TI - Leukotriene D4 binding and signal transduction in rat glomerular mesangial cells. AB - We examined the characteristics of [3H]leukotriene D4 (LTD4) binding to mesangial cells in culture. Binding is stereoselective, specific, saturable, and rapidly reversible. Two binding sites are recognized with dissociation constants and binding site densities at equilibrium of 2.2 and 16.8 nM and 1.1 x 10(4) and 3 x 10(4) binding sites per cell. LTD4, LTE4, (5R,6S)LTD4, LTB4, and the LTD4 receptor antagonist, SKF 104353, competitively inhibit radioligand binding in the following rank order of potency: LTD4 greater than LTE4 = SKF 104353 greater than (5R,6S)LTD4 greater than LTB4. LTD4 also induces time- and concentration dependent phosphoinositide hydrolysis in mesangial cells. Formation of inositol 1,4,5-trisphosphate (IP3) is maximal at 5 s, followed by a time-dependent increase in inositol monophosphate generation, and inhibited by 100-fold excess concentration of SKF 104353. Addition of LTD4 to mesangial cells is associated with an increase in intracellular pH and dose-dependent stimulation of [3H]thymidine incorporation and mitogenesis. Thus rat mesangial cells possess specific binding sites for LTD4, the activation of which stimulates IP3 formation and induces cellular alkalinization and mitogenic responses. These studies provide insight into the cellular basis for LTD4-mesangial cell interactions, which are of potential pathophysiological relevance during acute glomerular inflammatory injury. PMID- 2548404 TI - Sarcolemmal Ca2+ transport activities in cardiac hypertrophy caused by pressure overload. AB - To examine Ca2+ transport activities in sarcolemmal membrane in cardiac hypertrophy caused by pressure overload, rats were subjected to aortic banding for 28 days. Heart-to-body weight ratio was increased by 46% in aortic-banded animals in comparison with the sham-operated rats. Ouabain-sensitive Na+-K+ ATPase activity in sarcolemma (SL) from hypertrophied hearts was not different from that in the control preparation. The initial rate of Na+-dependent Ca2+ uptake in SL vesicles from the hypertrophied hearts was stimulated by 53% compared with the control vesicles. ATP-dependent Ca2+ uptake and Ca2+-stimulated adenosinetriphosphatase (ATPase) activities in SL from hypertrophied hearts were increased by 35%. The number of Ca2+ channels estimated by [5-methyl 3H]nitrendipine binding was decreased by 33% in SL from hypertrophied hearts. Total and individual phospholipid contents in the hypertrophied heart preparations were not different from those in the control, except that phosphatidylcholine and phosphatidylethanolamine contents were significantly increased. Sarcolemmal preparations from hypertrophied hearts from the 22-wk-old spontaneously hypertensive rats exhibited changes in Na+-Ca2+ exchange and Ca2+ pump activities (similar to those observed in banded hearts), whereas the Na+-K+ ATPase activity decreased, [3H]nitrendipine binding increased, and phospholipid contents were not different. Thus, although differences were defined between two types of hypertrophy, these results suggest alterations in the sarcolemmal Ca2+ transport activities that may serve as an adaptive mechanism for the removal of Ca2+ from the myocardial cells during the development of cardiac hypertrophy. PMID- 2548405 TI - Tissue-specific isoform regulation of Na+-K+-ATPase by thyroid hormone in ferrets. AB - We have shown previously that in the ferret heart there are two isoforms of Na+ K+-ATPase, alpha(+) and alpha, and that these isoforms undergo developmental changes. In the present study, we examine regulation of the isoenzymes by thyroid hormone, which is well known to increase activity of Na+-K+-ATPase in different tissue preparations. Ferrets were injected with L-thyroxine (T4) (0.5 mg/kg, sc) for 3 wk. The T4-treated ferrets gained 58 +/- 32 g body wt compared with 366 +/- 24 g for the control ferrets. Plasma 3,5,3'-triiodothyronine concentrations of the T4-treated animals increased about eightfold 16-18 h after the last injection. The number of alpha(+)- and alpha-subunits in heart homogenates estimated by [3H]ouabain binding was 3.5 +/- 0.1 and 2.5 +/- 0.1 pmol/mg protein, respectively [alpha(+)/alpha = 1.40]. In the T4-treated ferrets, there was no significant increase of the alpha(+)-isoform (3.2 +/- 0.2 pmol/mg protein), whereas the number of alpha-isoform increased significantly to 4.1 +/- 0.3 pmol/mg protein [alpha(+)/alpha = 0.78]. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of partially purified cardiac plasma membrane preparations reveals similar pattern of changes of the isoenzymes. In the kidney, however, there was no significant change in the number of alpha-isoform, which is the predominant isoform in the kidney, and T4 does not appear to induce synthesis of alpha(+)-isoform in the kidney. It is concluded that thyroid hormone induces both isoform- and tissue-specific regulation of the Na+-K+-ATPase alpha-subunits in the ferret. These specific regulations of the isoforms seem to suggest important physiological roles of the isoenzymes. PMID- 2548406 TI - Surgical removal of brown fat results in rapid and complete compensation by other depots. AB - Interscapular, scapular, and cervical brown adipose tissue (BAT) depots were removed from young male rats (BATX). These depots represented 40% of both the total dissectible mass and cytochrome oxidase content of BAT in these animals. Sham-operated and BATX rats were fed a low (8%) protein diet for periods of 9 or 16 days. Over both periods, body weight gain and energy intake, gain, expenditure, and efficiency were almost identical in sham and BATX groups. Four days after lipectomy the maximal thermogenic response (increase in O2 consumption) to norepinephrine (400 micrograms/kg sc) was 30% lower for BATX rats than for controls, but by day 13 this difference had disappeared. Nine days after lipectomy the total mass and cytochrome oxidase activity of the remaining dissectible BAT was comparable to that of the sham-operated controls, although the protein content was slightly reduced. The specific mitochondrial GDP binding (an index of thermogenic activity) was increased significantly in BATX rats, and total BAT mitochondrial GDP binding was no different from control values. At the end of the experiment (day 16), no regeneration of excised tissue had occurred, but the remaining BAT depots had shown almost complete compensation; the mass and the oxidative and thermogenic capacity of the total dissectible brown fat were virtually identical in both groups. PMID- 2548408 TI - Acidemia stimulates ACTH, vasopressin, and heart rate responses in fetal sheep. AB - Adrenocorticotropic hormone (ACTH), arginine vasopressin (AVP), and renin responses to hemorrhage are highly correlated to the hemorrhage-induced decreases in arterial pH. The present study was designed to test the responses of these three systems to acute fetal acidemia, produced by intravenous infusion of H+. HCl was infused into chronically catheterized fetal sheep at rates of 0.02 (n = 5), 0.10 (n = 6), and 0.50 (n = 5) meq/min. Infusions at rates of 0.10 and 0.50 meq/min significantly decreased fetal arterial pH and increased arterial PCO2. Fetal heart rate and plasma concentrations of ACTH, cortisol, and AVP were significantly increased during infusion of HCl at 0.5 meq/min. Neither fetal plasma renin activity nor fetal arterial blood pressure was significantly altered by any of the infusions. The results of these experiments suggest that fetal ACTH, AVP, and heart rate are stimulated by decreases in arterial pH and/or increases in arterial PCO2. We speculate that these responses are chemoreceptor mediated, although we cannot distinguish the apparent relative roles of peripheral and central chemoreceptors on the basis of the present study. PMID- 2548407 TI - Cold-induced changes in uncoupling protein and GDP binding sites in brown fat of ob/ob mice. AB - The acute and chronic effects of cold exposure on the thermogenic activity and capacity of brown fat of obese (ob/ob) and lean mice have been examined. After 1 h at 13 degrees C, mitochondrial GDP binding increased in warm-acclimated (28 degrees C for 3 wk) lean mice, and increases in mitochondrial mass and the specific mitochondrial concentration of uncoupling protein were evident at 24 h. Long-term cold exposure led to further progressive increases in the specific mitochondrial concentration and total tissue content of uncoupling protein. The acute increase in GDP binding was due to an unmasking of binding sites on the protein and was accompanied by a parallel increase in the acetate-induced swelling of the mitochondria. In contrast to lean animals, ob/ob mice did not exhibit an increase in GDP binding or unmasking of binding sites during the first 24 h of cold exposure; unmasking was, however, evident by 3 days. A delay in the stimulation by cold of both the specific mitochondrial concentration of uncoupling protein (not increased until 3 days) and the mitochondrial content of brown fat (not increased until 8 days) occurred in the obese mice. Acute warm exposure (28 degrees C) of cold-acclimated (13 degrees C for 3 wk) mice led to a reduction in GDP binding in both lean and obese animals because of a rapid masking of binding sites. Mitochondrial content and the specific mitochondrial concentration of uncoupling protein fell progressively in both groups of mice with continued warm exposure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548409 TI - Catecholamine activity in paraventricular hypothalamus after hemorrhage in cats. AB - Temporal changes in catecholamine activity within the paraventricular nucleus (PVN) of the hypothalamus were assessed using in vivo voltammetry after moderate hemorrhage in chloralose-anesthetized cats. Oxidation current was measured with carbon microelectrodes, and the change at +250 mV was used as an estimate of catecholamine activity. The magnitude and directional change in catecholamine activity was assessed for each site during the initial 3 min of blood loss. Of 62 recording sites, 45 sites were located along the rostrocaudal extent of the medial PVN. Fifteen of these 45 sites exhibited an increase, 15 others exhibited a decrease, and the remaining 15 sites exhibited no change in catecholamine activity. The magnitude of change in oxidation current was independent of the magnitude of the decrease in arterial pressure after hemorrhage. Twelve of the 15 sites that exhibited increases in catecholamine activity were located in the caudal PVN along its dorsomedial aspect, whereas the location of sites that exhibited decreases in catecholamine activity were more widely distributed throughout the nucleus. These data are consistent with the hypothesis that an increase in the release of catecholamines within the caudal PVN in response to hemorrhage is facilitatory for the release of adrenocorticotropin and/or vasopressin. PMID- 2548410 TI - Effects of norepinephrine and denervation on brown adipose tissue in Syrian hamsters. AB - Daily injections of either 0.8 or 3.2 mg norepinephrine (NE)/kg for 2 wk failed to stimulate brown adipose tissue (BAT) growth, GDP binding, or cytochrome-c oxidase activity (COA) in Syrian hamsters (Mesocricetus auratus). However, a single injection of 1.6 mg NE/kg produced a small (23%) but significant acute increase in BAT GDP binding without affecting COA. Thus there is some loss of sensitivity to NE with chronic treatment in Syrian hamsters. Unilateral sympathectomy by surgical denervation of the interscapular BAT (IBAT) resulted in decreased GDP binding and COA in the denervated pad. Chronic NE treatment in hamsters with denervated IBAT only partially reversed the denervation-induced decreases in GDP binding and COA. It therefore appears that NE is not solely responsible for the maintenance and stimulation of thermogenic activity and COA in Syrian hamster BAT. Denervation of IBAT also resulted in elevated levels of lipoprotein lipase (LPL) in this tissue, a surprising finding since brown and white adipose tissue LPL activity were both stimulated by chronic NE treatment. Therefore, although NE has a stimulatory effect on LPL activity, the primary influence of the neural input to IBAT on this enzyme is inhibitory. These data exemplify dramatic differences between rats and hamsters in the mechanisms controlling BAT thermogenesis and white and brown adipose tissue LPL activity. PMID- 2548411 TI - Control of canine ACTH by corticosteroids: an integral feedback effect of steroids. AB - These experiments were designed to test whether the pattern of change in plasma corticosteroid or the total corticosteroid dose is important in determining the degree of inhibition of adrenocorticotropic hormone (ACTH) responses to stress by corticosteroid intermediate-delayed feedback. Five conscious dogs were studied. The ACTH response to induced hypoglycemia was measured after no prior corticosteroid feedback signal or after a corticosteroid feedback signal produced by infusion, two bolus injections, or three bolus injections of cortisol and corticosterone. The total corticosteroid dose (45 micrograms/kg) and the total interval of steroid treatment (60-30 min before hypoglycemia) were the same in all three cases of corticosteroid treatment. Changes in plasma glucose concentration during induced hypoglycemia were not altered by corticosteroid treatment. The plasma ACTH response to hypoglycemia was inhibited by all three patterns of treatment with corticosteroids. The inhibition of ACTH response was not significantly altered among the patterns of treatment with corticosteroids. The data suggest that the integrated (total) or the mean change in plasma corticosteroid concentration over time determines the degree of inhibition of stimulated ACTH in this time domain. PMID- 2548412 TI - Hyalinizing trabecular adenoma. PMID- 2548413 TI - [The serum concentration of specific beta 1-glycoprotein in trophoblastic disease]. AB - Twelve women with hydatidiform mole were studied between 12 and 20 weeks gestation. The level of SP1 was determined by radioimmunologic method. The author found statistically significant increase of SP1 over 90th percentile of gestational age (p less than 0.001). Examining the level of SP1 after evacuation of hydatidiform mole it was established that SP1 was eliminated more quickly than human chorionic gonadotropin (HCG) in urine during early pregnancy. Quite the contrary was discovered in women with advanced pregnancy, in whom the concentration of SP1 disappeared more slowly, while HCG was eliminated for a shorter time. The observed dissociation in elimination of SP1 and HCG showed that the combined examination of HCG and SP1 provided better information in the follow up of women with trophoblastic disease. The author think that treatment of trophoblastic disease should continue till complete elimination both of HCG and SP1. PMID- 2548414 TI - [Gamma-camera scintigraphy and radioimmunological study of the hormones of the hypophyseal-gonadal axis to evaluate testicular function in patients with hypogonadism and infertility]. AB - Gamma-camera and radioimmunological study of hormones of hypophyseal-gonadal axis were carried out on 34 patients with hypogonadism and infertility after successfully performed orchidopexy. The results from this study showed that gamma camera scintigraphy was exact and noninvasive method for determining the structure and function of testes after operative intervention. The results of the established hormonal deviations in the secretion of hormones of hypophyseal gonadal axis are sure and early criterium for diagnosis of infertility in these patients. They could indicate the therapeutic management in these patients. PMID- 2548416 TI - Blockade of sciatic nerve branches relieves sciatic radicular pain. PMID- 2548415 TI - Intrathecal clonidine suppresses noxiously evoked activity of spinal wide dynamic range neurons in cats. AB - The analgesic effectiveness of perispinal clonidine administration prompted us to evaluate clonidine effects on spinal dorsal horn wide dynamic range neurons. Intrathecal clonidine produced a dose-dependent (10 and 30 micrograms), yohimbine reversible suppression of noxiously evoked activity in decerebrate, spinal cord transected cats. In addition, combining ineffective intrathecal doses of morphine (25 micrograms) and clonidine (5 micrograms) produced statistically significant, reversible suppression of noxiously evoked activity. The time course of suppression was similar to that observed behaviorally. These results support the role of spinal alpha 2-adrenergic receptors in clonidine analgesia. PMID- 2548417 TI - Antigenic and restriction enzyme analysis of isolates of Campylobacter fetus subsp venerealis recovered from persistently infected cattle. AB - Thirty-two isolates of Campylobacter fetus subsp venerealis were obtained from 1 bull and 4 heifers with experimentally induced infection. When whole-cell antigens of isolates were cross titrated with antisera to the infecting strain, isolates from 3 heifers had limited antigenic variation, whereas whole-cell antigens of isolates from 2 cattle (the bull and a heifer) differed serologically from those of the infecting strain. Changes were detected specifically in 6 heat labile antigens. Of the 6 heat-labile factors evaluated, all were initially present on the infecting parent strain, but not on early isolates obtained from 4 of the 5 cattle. Restriction enzyme analysis revealed minor variation in the DNA fingerprints of isolates obtained from individual cattle, thus implying stability of the Campylobacter genome once persistent infection is established. Isolates with identical restriction enzyme patterns expressed different heat-labile antigens. Correlation could not be found between the DNA electrophoretic pattern and the expression of heat-labile antigens. PMID- 2548418 TI - Transmission of a vaccinal strain of infectious bovine rhinotracheitis virus from intranasally vaccinated steers commingled with nonvaccinated steers. AB - Ninety-seven feeder steers, averaging 7 months of age, were allotted to 3 groups. Group I (n = 33) was vaccinated intranasally with an infectious bovine rhinotracheitis virus (IBRV) vaccine on postinoculation day (PID) 0; group II (n = 31) was not vaccinated on PID 0 but was commingled with group I; and group III (n = 33) served as controls housed in the same facility, but was physically separated from groups I and II. On PID 20, all steers were given a modified-live IBRV vaccine IM. Virus isolation attempts from nasal swab specimens collected on PID 10 resulted in IBRV isolation from 19 (57.6%) of group I, 4 (12.9%) of group II, and 0 of group III. By PID 20, geometric mean titer for serum antibody to IBRV had increased in group I but had decreased in groups II and III. By PID 40, geometric mean titer for serum antibody to IBRV had increased in the 3 groups in response to IM vaccination given on PID 20. Seemingly, transmission of a vaccinal strain of IBRV to nonvaccinated steers did not take place at a frequency that elicited a humoral immune response similar to that of vaccinated steers. PMID- 2548419 TI - Specificity of neutralizing and precipitating antibodies induced in healthy calves by monovalent modified-live bovine viral diarrhea virus vaccines. AB - Serum was obtained at weekly intervals after vaccination of 6 healthy calves with either of 2 commercially available monovalent modified-live bovine viral diarrhea (BVD) virus vaccines. Detectable neutralizing antibodies to each of 10 cytopathic and 10 noncytopathic isolates of BVD virus were produced by 1 or more of the calves by 14 days after vaccination, but no calf produced detectable neutralizing antibodies to all 20 BVD viruses. At that time, precipitating antibodies against viral-induced polypeptides of approximately 115,000; 80,000; 56,000; 48,000; 39,000; and 25,000 daltons were detected in sera from some calves. Also at that time, specificity of the antibodies for polypeptides of certain viruses was detected. At 21 days after vaccination, each calf produced neutralizing antibodies to all 20 BVD viruses. At that time, precipitating antibodies to each of the aforementioned viral induced polypeptides were detected in serum from each calf. Precipitating antibodies to viral induced polypeptides of 61,000 and 37,000 daltons were detected in samples of sera obtained from some calves at 42 days after vaccination. PMID- 2548420 TI - Comparative virulence of two porcine group-A rotavirus isolates in gnotobiotic pigs. AB - The virulence of 2 porcine group-A rotavirus isolates was compared. Forty hysterotomy-derived 3-day-old gnotobiotic pigs were inoculated orally with 2 ml of intestinal homogenate containing either the Ohio State University (OSU) or the South Dakota State University (SDSU) strain of porcine rotavirus or were inoculated with medium only. Clinical signs of disease, body weight, distribution of viral antigen, fecal excretion of virus, and histologic lesions (observed by light and scanning electron microscopy) were determined. Morphometric measurements of villi and crypts were made. In pigs inoculated with OSU or SDSU strains, diarrhea began at postinoculation hours (PIH) 19 to 48 and PIH 24 to 54, respectively. None of the virus-infected pigs died as a consequence of infection and all had similar clinical signs of disease, body weight changes, and virus shedding patterns, regardless of the strain of rotavirus with which they were infected. Microscopic findings in the small intestine of virus-infected pigs were similar, except that the SDSU strain caused more severe villus atrophy and villus fusion in the duodenum at PIH 72 and 168 than was associated with the OSU strain. Viral antigen in the small intestine of pigs infected with either virus was observed by use of immunofluorescence at PIH 24 and 72, but was seldom seen at PIH 168. PMID- 2548421 TI - Immunohistochemical staining and radionuclide imaging of canine tumors, using a monoclonal antibody recognizing a synthetic carbohydrate antigen. AB - The in vitro and in vivo binding of a monoclonal antibody (MAB) that recognizes a tumor-associated carbohydrate antigen was studied in dogs. Monoclonal antibody 155H.7 was raised in response to innoculation of mice with beta-galactose(1 3)beta N-acetylgalactosamine conjugated to human serum albumin. Avidin-biotin complex immunohistochemical staining of cryostat sections of normal and neoplastic canine tissue specimens revealed heterogenous binding of MAB 155H.7 to the cells of many canine mammary and lung carcinomas and homogenous staining of many sarcomas, including osteogenic sarcoma. In addition, there was variable staining of a variety of normal tissues including some ductual epithelium, peripheral nerve fibers, and some endothelial cells and fibroblasts. Immunoscintigraphy with 131I-labeled MAB 155H.7 was used to study the in vivo distribution of the antibody. The 131I-labeled MAB 155H.7 was administered to 1 clinically normal dog, 7 dogs with osteogenic sarcoma, 1 dog with undifferentiated sarcoma, and 2 dogs with mammary tumor. Scintigraphy revealed concentration of radioactivity in 8 of 10 tumor sites within 24 hours after MAB administration. The ratio of 131I in tumor sites to 131I in the surrounding normal tissues, compared with the similar ratio of 99mTc-labeled erythrocytes ranged from 1.1 to 4.3, in tumor vs normal tissue with a mean value of 2, confirming tumor localization of the radiolabeled MAB in excess of that associated with enhanced tumor vascularization. PMID- 2548422 TI - Role of leukotriene B4 in the pathogenesis of Klebsiella pneumoniae-induced bovine mastitis. AB - Mastitis was induced in 4 lactating cows by inoculation of Klebsiella pneumoniae (10(7) organisms/ml) via the teat canal. Sterile isotonic saline solution (1 ml) was instilled into designated control quarters via the teat canal. Changes in milk leukotriene B4 and C4 (LTB4, LTC4) concentrations, milk somatic cell counts, and milk bovine serum albumin concentration were monitored over a 24-hour postinoculation period. Milk LTB4 concentration before inoculation in control quarters and quarters later to be infected was 376 +/- 45 and 326 +/- 56 pg/ml of milk, respectively. A significant (P less than 0.05) increase in milk LTB4 concentration in the infected quarters was first observed at postinoculation hour 6, and milk LTB4 concentration in infected quarters generally remained significantly high through postinoculation hour 14. Thereafter, milk LTB4 concentration in infected quarters was not significantly different from the concentration in control quarters. Measurable amounts of LTC4 were not detected in the milk of either control or infected quarters. Milk bovine serum albumin concentration in the infected quarters generally was high throughout the study, as were milk somatic cell counts. The results of this study suggested that LTB4 contributes to the pathogenesis of bovine mastitis. PMID- 2548423 TI - Effect of canine parvovirus on erythroid progenitors in phenylhydrazine-induced regenerative hemolytic anemia in dogs. AB - The effects of canine parvovirus (CPV) infection in dogs with hemolytic anemia was compared with the clinical effects of human parvovirus-induced aplastic anemia in human beings with chronic regenerative anemias. Phenylhydrazine was used to induce a transient, severe, hemolytic anemia in dogs to evaluate the effects of CPV infection on rapidly dividing bone marrow precursors. Erythrocyte colony-forming unit bone marrow cultures and cytologic examination of bone marrow were used to determine the effects of CPV infection on erythroid bone marrow precursors. The induced hemolytic anemia regenerated rapidly and although the bone marrow was infected, it was determined that CPV infection did not induce a detectable decrease in erythroid progenitors in dogs with severe hemolytic anemia. PMID- 2548424 TI - Trypsin treatment of bovine ova after in vitro exposure to vesicular stomatitis virus. AB - Preimplantation bovine ova were exposed in vitro to vesicular stomatitis virus, Indiana serotype, to document adherence of the virus to the zona pellucida. To determine the efficacy of this treatment, some of the ova were treated with trypsin after exposure to the virus. Vesicular stomatitis virus was isolated from 5 of 10 groups of zona pellucida-intact ova after 12 sequential washes without trypsin treatment. Vesicular stomatitis virus was also isolated from 4 of 11 groups of zona pellucida-intact ova after trypsin treatment. PMID- 2548425 TI - Effect of theophylline and adenosine on eosinophil function. AB - Eosinophils may play a critical role in asthma and bronchial hyperresponsiveness, yet the effect of theophylline on their function is not certain. We have examined the effects of theophylline on opsonized zymosan-induced superoxide anion (O2-) release from guinea pig eosinophils harvested from the peritoneal cavity and from human eosinophils obtained by differential centrifugation of blood from patients with peripheral eosinophilia. Theophylline at high concentration (10(-3) M) inhibited O2- release by 27.6 +/- 9.4% (mean +/- SEM, p less than 0.05), whereas at clinically relevant concentrations (10(-6) and 10(-5) M), it significantly potentiated this by 26.8 +/- 9.9% (p less than 0.05) and 36.9 +/- 6.3% (p less than 0.01), respectively. 8-phenyltheophylline (10(-7) to 10(-3) M), which like theophylline inhibits adenosine receptors but does not inhibit phosphodiesterase activity, produced potentiation at all concentrations. Preincubation of eosinophils with adenosine deaminase (0.1 U/ml) enhanced O2- release by 72.4 +/- 15.2% (p less than 0.01), whereas addition of adenosine (3 x 10(-8) to 10(-6) M) reversed the potentiation induced by theophylline (10(-5) M) in a concentration dependent manner. Inhibition was greater with the A2-selective analog N ethylcarboxamide adenosine than the A1-selective analog phenylisopropyladenosine, suggesting that A2-receptors are involved. In human eosinophils we have demonstrated a similar effect of theophylline and adenosine on O2- release. Our results indicate that therapeutic concentrations of theophylline may potentiate eosinophil activation in vivo by competing with circulating adenosine for eosinophil A2-receptors. This would be consistent with the lack of effect of theophylline on bronchial hyperresponsiveness, which may be related to eosinophilic inflammation. PMID- 2548426 TI - Bronchiolitis in adults. A reversible cause of airway obstruction associated with airway neutrophils and neutrophil products. AB - In the past 4 yr, 16 adult patients were identified who had accelerated onset of a severe respiratory disorder (usually obstructive in nature) that was clinically distinct from the more commonly encountered chronic obstructive disorders (e.g., chronic bronchitis, emphysema, asthma, bronchiectasis, cystic fibrosis, and alpha 1-antitrypsin deficiency). These patients, termed patients with "bronchiolitis," underwent pulmonary function testing, bronchoscopy with bronchoalveolar lavage (BAL), and open lung biopsy. Although lung biopsy findings varied somewhat among the patients, each biopsy contained a prominent component of bronchiolitis. Pulmonary function testing and BAL were also repeated after 3 months of treatment with oral prednisone (1 mg/kg/day). Initial BAL neutrophil percentages were significantly higher in the bronchiolitis group (54 +/- 10%) than in smokers with chronic bronchitis (3.9 +/- 1.0%) or in normal nonsmoking volunteers (0.8 +/- 0.5%) (p less than 0.01, both comparisons). Eleven of 15 patients with bronchiolitis had significant improvement (greater than or equal to 15% increase in FEV1) in their lung function after prednisone treatment. Furthermore, this "responder" subgroup had a significant reduction in BAL neutrophil percentages after treatment with prednisone (46 +/- 15% to 6 +/- 3%, p less than 0.05). Finally, the neutrophil products collagenase and myeloperoxidase were detected in the BAL fluid of patients with bronchiolitis. These findings suggest a central role for the neutrophil in the pathogenesis of bronchiolitis and emphasize the utility of BAL in the identification of these patients. PMID- 2548427 TI - Response of glucagonomas to surgical excision and chemotherapy. Report of two cases and review of the literature. AB - The glucagon-producing pancreatic tumors or glucagonomas are among the rarest forms of islet cell tumors; most are malignant and usually produce a definite clinical syndrome. Mild diabetes mellitus, weight loss, and anemia usually accompany the syndrome. However, only the presence of a peculiar cutaneous rash (necrolytic migratory erythema) and the finding of hyperglucagonemia on assay are reliable diagnostic features of the syndrome. Selective, celiac axis arteriography is the most valuable preoperative technique for localizing these neoplasms and their common liver metastases. Immunohistochemical and ultrastructural examinations are particularly helpful in defining the tumor cell nature (alpha-2 islet cell) and the peptide content (glucagon). When the tumor is benign (less than 30%), complete operative removal results in lasting cure; for malignant forms, surgical therapy is mainly palliative, and adjunctive chemotherapy should be administered. In this report, the importance of clinical recognition and operative and chemotherapeutic responses is illustrated in two patients. In each case, the characteristic dermatitis, diabetes mellitus, weight loss, anemia, and elevated plasmatic glucagon were present. Both patients had their tumors localized by selective angiography and underwent operative removal of the primary pancreatic lesion. In the case of benign glucagonoma, surgical excision was curative. In the malignant one, cytoreductive surgery plus adjunctive chemotherapy (dimethyltriazenomidazole-carboxamide resulted in prolonged survival and significant clinical improvement. Follow-up with serum glucagon assay has been useful in monitoring recurrence. PMID- 2548428 TI - Synchronous hepatocellular carcinoma and adenocarcinoma of the colon. AB - It is not uncommon for patients with colon cancer to have a second primary neoplasm. The reported incidence of synchronous colon tumors is approximately about 3 to 4 per cent. Colon cancer also may be associated with a synchronous extra-colonic neoplasm; this incidence is less well defined and certainly is less emphasized in the literature. The authors recently treated a patient who had a synchronous hepatoma and adenocarcinoma of the colon. This appears to be the first report of these tumors presenting synchronously. The patient had both tumors resected and has no evidence of recurrence ten months later. This case illustrates the need to seek both colonic and extra-colonic synchronous tumors in patients discovered to have a colon cancer. PMID- 2548429 TI - The acquired immunodeficiency syndrome (AIDS) dementia complex. AB - The acquired immunodeficiency syndrome (AIDS) dementia complex is a frequent and devastating complication of infection with human immunodeficiency virus-type 1 (HIV-1). Features of the AIDS dementia complex include decreased memory, the inability to concentrate, apathy, and psychomotor retardation. Typical neuropathologic findings include gliosis, focal necrosis of neurons, perivascular inflammation, formation of microglial nodules, multinucleated giant cells, and demyelination. That HIV-1 is the direct cause of this neurologic syndrome is strongly supported by the available evidence. In addition, several studies have identified the monocyte-macrophage as the predominant cell type in the brain infected with HIV-1. However, the mechanisms by which the infected monocytes macrophages mediate neurologic dysfunction and destruction have not been elucidated. PMID- 2548431 TI - [Hepatocellular carcinoma in cirrhosis. The surgeon's point of view]. PMID- 2548430 TI - [A new experimental model of cecal amebiasis in rats]. AB - The young rat was used as an experimental model of caecal amebiasis by implanting, without injury of the caecum, a suspension of E. histolytica trophozoites gnotoxenic strain Rahman of high virulence. The degree of pathogenicity is demonstrated by the macroscopic aspect of caecum, by the density of trophozoites and by an anatomopathologic examination. As a correlation exits between all these macroscopic parameters and histology, a degree of illness is fixed which include several macro- and microscopic criteria. This model enables to observe the different phases of cicatrization and of recovering of young rats and so to appreciate the activity of antiamebic agents. PMID- 2548432 TI - Synergism of prostaglandin E2 plus TNF in induction of differentiation of human monocytoid leukemic U-937 cells. AB - Prostaglandin E2 (PGE2) induced differentiation of human monoblastic leukemia U 937 cells into cells with macrophage characteristics. PGE2 synergistically increased the differentiation, of U-937, ML-1 and HL-60 cells when combined with TNF. PGE1 and PGF2 also induced differentiation of U-937 cells; however, PGD2, deoxy-delta 9,12-13, 14-dihydro-PGD2 (delta 12-PGJ2), and PGI2 did not induce differentiation, either alone or in combination with TNF. PGE2 changed the dissociation constant of TNF for its receptors on U-937 cells only marginally, but it approximately doubled the number of binding sites. PMID- 2548433 TI - Cytoskeletal protein and GTP-ase profiles differ depending on the degree of transformation of fibroblasts. AB - Four fibroblast cell lines of differing rates of cell division (1 normal and 3 transformed to differing degrees) were cultured in 10% FCS and 90% DMEM. The cultures were maintained for 72 hours, either dosed with C18 fatty acids or retained as controls, and cell viability was then assessed using the Trypan Blue exclusion method. The cytoskeletal proteins were then prepared, and the protein and GTP-ase profiles determined. No changes in profile were detected within a cell type whether dosed with fatty acid or not. Between cell types, whilst total cytoskeletal protein remained constant, distinct changes were observed in the PAGE. The more transformed the cells, the fewer protein bands detected, especially those with masses of 50-60kD. Tubulin has sub-units with masses in this range, and also expresses GTP-ase activity. This activity decreased in parallel to the changes in protein profiles. PMID- 2548434 TI - Histochemically demonstrable phosphotyrosyl-protein phosphatase in normal human breast, in benign breast diseases and in breast cancer. AB - The activity of phosphotyrosyl-protein phosphatase enzyme was investigated by a histochemical method in the normal human breast and in breast diseases in order to evaluate its possible significance in the genesis and in the growth of benign and malignant epithelial proliferative. In normal human breast tissue only a weak enzyme activity was present. The activity was elevated in benign disease in actively proliferative lesions and in 71% of the cases of breast cancers. When enzyme activity of breast cancers was compared with the content of receptors for epidermal growth factor and insulin-like growth factor-I, no association was found. It is concluded that phosphotyrosyl-protein phosphatase is increased in actively proliferating human breast diseases. Thus the putative increase in phosphotyrosyl-proteins mediating benign and malignant epithelial proliferations is rather caused by an increase in protein-tyrosine kinase activity than by a decrease in phosphotyrosyl-protein phosphatase activity. PMID- 2548435 TI - Expression of ras and myc oncogenes in human hepatocellular carcinoma and non neoplastic liver tissues. AB - An immunohistochemical assay was used to assess expression of ras p21 and myc p62 oncogene products in human hepatocellular carcinoma (HCC) and non-neoplastic liver tissues. The monoclonal antibodies Y13 259 and Myc1-9E10, specific for ras p21 and myc p62 oncoproteins, were employed on paraffin-embedded sections. Most HCCs showed enhanced ras p21 and myc p62 expression, as indicated by staining intensity. Cirrhotic livers revealed increased myc p62 and occasionally increased ras p21 expression. HBsAg+ hepatocytes showed intense immunostaining for ras p21. Fibrotic, cholestatic, fetal and normal adult liver did not present enhancement of oncoprotein production. We suggest that combined over-expression of ras and myc oncoproteins may be important for the malignant phenotypic alteration in human HCC. PMID- 2548436 TI - Changes of serum iron transferrin and copper ceruloplasmin in rats given Cu(II)2 (acetylsalicylate)4 during acute inflammation. AB - Modifications of serum levels of iron transferrin and copper ceruloplasmin after acute inflammation by carrageenan and treatment with acetyl salicylic acid [ASA] or Cu(II)2(acetylsalicylate)4 [Cu(II)2(AS)4] were studied in the rat by EPR spectroscopy. Furthermore, the ulcerogenic potential of the two drugs was investigated after a single high oral dose. Our results indicate that Cu(II)2(AS)4 is more effective than ASA in limiting the inflammation provoked by the phlogogen. In these conditions the iron(III) non-heme and copper(II) ceruloplasmin concentration in serum was modified either during inflammation or after treatment with antiphlogistic agents: in carrageenan-injected rats the level of serum iron(III) non-heme was found to be very low, while the copper(II) ceruloplasmin concentration was partially reduced. On the other hand, after the pharmacological treatments, no changes of iron transferrin were observed and the concentration of copper ceruloplasmin was increased. With regard to their ulcerogenic effect, ASA appeared to be more irritating for gastric mucosa than Cu(II)2(AS)4. PMID- 2548437 TI - In vitro studies on tin-dithiocarbamates. AB - Organotin compounds have been studied for their biological properties, whereas very little is known about the effects of tin-inorganic derivatives on biological systems. Owing to our interest in the dithiocarbamate moiety in relation to metal redistribution in the body and to its biological import, we performed in vitro cytotoxicity experiments on dithiocarbamates. Preliminary results obtained with five tin-dithiocarbamates, where DDTC is the diethyldithiocarbamate anion, indicate some very promising compounds. PMID- 2548438 TI - Different binding to DNA of enantiomeric platinum complexes assessed by inhibition of restriction enzyme activity. AB - The ability of enantiomeric platinum complexes to block the action of selected restriction enzymes has been investigated. The complexes [PtCl2(DAC)], [PtCl2(DAB)] and [PtCl2(DAP)] (DAC = 1,2-diamminocyclohexane; DAB = 2,3 diamminobutane; DAP = 1,2-diamminopropane) exhibit a guanine-cytosine preference in accord with previous results on cis-[PtCl2(NH3)2] (cis-DDP). The extent of inhibition, however, is significantly different for the different isomers; the R,R form is more active than the others at short incubation time, as the time becomes longer, the differences among isomers level off. It also appears that cis DDP is more active than [PtCl2(DAC)] in blocking the Cfo I enzyme, though it shows a preference for the G-X-G sequences. PMID- 2548439 TI - Cloning and characterization of a DNA gyrase A gene from Escherichia coli that confers clinical resistance to 4-quinolones. AB - Nalidixic acid, enoxacin, and other antibacterial 4-quinolones inhibit DNA gyrase activity by interrupting DNA breakage and reunion by A subunits of the A2B2 gyrase complex. Despite their clinical importance, the mode of quinolone action and mechanisms of resistance are poorly understood at the molecular level. Using a DNA fragment enrichment procedure, we isolated the gyrA gene from a uropathogenic Escherichia coli strain that encodes a gyrase A protein cross resistant to a variety of quinolones. When complemented with gyrase B subunit, the purified A protein reconstituted DNA supercoiling activity approximately 100 fold more resistant to inhibition by enoxacin than the susceptible enzyme and failed to mediate quinolone-dependent DNA cleavage. Nucleotide sequence analysis revealed that the gene differed at 58 nucleotide positions compared with the K-12 gyrA sequence. The 875-amino-acid residue-resistant gyrase A protein differed at three positions from its wild-type E. coli K-12 counterpart: tryptophan, glutamate, and serine replaced serine, aspartate, and alanine residues at positions 83, 678, and 828, respectively. By genetic analysis of chimeric gyrA genes in a gyrA(Ts) background, we showed that the Ser-83----Trp mutation in the gyrase A protein was solely responsible for high-level bacterial resistance to nalidixic acid and fluoroquinolones. PMID- 2548440 TI - Human immunodeficiency virus inhibition is prolonged by 3'-azido-3' deoxythymidine alternating with 2',3'-dideoxycytidine compared with 3'-azido-3' deoxythymidine alone. AB - The inhibition of the lymphadenopathy-associated virus strain of human immunodeficiency virus (HIV) by alternating regimens of two dideoxynucleosides, 3'-azido-3'-deoxythymidine (AZT) (zidovudine) and 2',3'-dideoxycytidine (ddC), was determined in CEM cells. Cultures infected with virus for 2 h were treated with clinically achievable concentrations of AZT, ddC, or a 3-day-alternating regimen of AZT and ddC. Media were completely changed every 3 days and replaced with antiviral agent, and virus production was assayed by p24 antigen and virus specific DNA. Cells treated with no antiviral agent exhibited breakthrough infection by day 6 in culture, whereas cells treated with 0.1, 1.0, or 3.0 microM AZT had a prolonged time to viral breakthrough. For each regimen of AZT alternating with 0.05 or 0.1 microM ddC, there was consistently prolonged HIV inhibition compared with continuous treatment with AZT alone. The viral suppression achieved with the alternating combinations required AZT as well as ddC and was superior to 3 days of treatment with ddC alternating with 3 days of no antiretroviral treatment. Levels of unintegrated HIV DNA paralleled the detection of p24 antigen, with the most prolonged inhibition of virus-specific DNA occurring with AZT alternating with ddC (compared with all regimens except continuous treatment with ddC). These data suggest that alternating regimens of AZT and ddC not only might decrease toxicity associated with the two drugs but may prove to be more efficacious than AZT alone. PMID- 2548441 TI - Resistance emerging after pefloxacin therapy of experimental Enterobacter cloacae peritonitis. AB - Resistance emerging after pefloxacin therapy was investigated in an experimental Enterobacter cloacae infection. Mice were inoculated intraperitoneally (mean inoculum, 0.9 X 10(8) CFU) with one of four strains initially susceptible to quinolones and treated with a single 25-mg/kg dose of pefloxacin. This therapy produced a net decrease of bacterial counts in the peritoneal fluid, but with the of the isolates, posttherapy (PT1) strains emerged with decreased susceptibilities to quinolones (4- to 1,024-fold), to the structurally unrelated antibiotics (4- to 16-fold) chloramphenicol and trimethoprim, and sometimes to tetracycline and beta-lactam compounds. In a second set of experiments, new mice were similarly infected with PT1 strains and treated with up to five 25-mg/kg doses of pefloxacin. Compared with parent isolates, PT1 strains produced similar disease and peritoneal bacterial count in the control animals. In treated mice posttherapy (PT2) strains emerged that showed 8- to 64-fold increases in quinolone MICs compared with the PT1 strains inoculated. All PT1 and PT2 strains showed altered outer membrane protein patterns, principally marked by a decreased 37,000-molecular-weight band generally accompanied by an increased 42,000 molecular-weight band. Whole cells from all PT1 and PT2 strains, exposed to [14C]pefloxacin for 15 to 60 s, bound significantly less radioactivity than the corresponding parent strains. After partial purification, DNA gyrase extracted from the most resistant isolates (one PT1 and the PT2 strains) showed a 100- to 450-fold 50% inhibitory concentration increase for pefloxacin. Altogether, pefloxacin can select in vivo two types of resistant strain, one with only decreased permeability and another with decreased permeability combined with altered DNA gyrase. PMID- 2548443 TI - Sputum levels of fluconazole in humans. AB - We measured fluconazole levels in sputum samples obtained from 11 bronchiectatic volunteers at 4 and 24 h after a single oral dose of 150 mg of fluconazole. Levels in sputum were similar to levels in plasma at both times. PMID- 2548442 TI - In vitro susceptibilities of Entamoeba histolytica to azithromycin, CP-63,956, erythromycin, and metronidazole. AB - Current therapy of Entamoeba histolytica infection requires use of multiple agents effective at different body sites, including the intestinal lumen, intestinal tissue, and liver. Azithromycin and CP-63,956, new extended-half-life macrolides which reach high levels in tissue, exhibit in vitro antiamebic activity at 18 or 48 h of incubation at concentrations comparable to that of erythromycin and slightly higher than that of metronidazole. Azithromycin and CP 63,956 have the potential to be useful therapeutic agents for all types of E. histolytica infection. PMID- 2548444 TI - In vitro activity of SK&F 104662, a new glycopeptide antibiotic. AB - The in vitro activity of SK&F 104662, a new glycopeptide antibiotic, against gram positive bacteria was evaluated. Activity was comparable to those of teicoplanin and vancomycin against most organisms. SK&F 104662 inhibited diphtheroids at concentrations of less than or equal to 0.5 microgram/ml. Addition of human serum to the test medium lowered the inhibitory activity of this glycopeptide against some organisms by as much as eightfold. PMID- 2548445 TI - Differential depuration of poliovirus, Escherichia coli, and a coliphage by the common mussel, Mytilus edulis. AB - The elimination of sewage effluent-associated poliovirus, Escherichia coli, and a 22-nm icosahedral coliphage by the common mussel, Mytilus edulis, was studied. Both laboratory-and commercial-scale recirculating, UV depuration systems were used in this study. In the laboratory system, the logarithms of the poliovirus, E. coli, and coliphage levels were reduced by 1.86, 2.9, and 2.16, respectively, within 52 h of depuration. The relative patterns and rates of elimination of the three organisms suggest that they are eliminated from mussels by different mechanisms during depuration under suitable conditions. Poliovirus was not included in experiments undertaken in the commercial-scale depuration system. The differences in the relative rates and patterns of elimination were maintained for E. coli and coliphage in this system, with the logarithm of the E. coli levels being reduced by 3.18 and the logarithm of the coliphage levels being reduced by 0.87. The results from both depuration systems suggest that E. coli is an inappropriate indicator of the efficiency of virus elimination during depuration. The coliphage used appears to be a more representative indicator. Depuration under stressful conditions appeared to have a negligible affect on poliovirus and coliphage elimination rates from mussels. However, the rate and pattern of E. coli elimination were dramatically affected by these conditions. Therefore, monitoring E. coli counts might prove useful in ensuring that mussels are functioning well during depuration. PMID- 2548446 TI - Protection of rainbow trout against vibriosis and furunculosis by the use of attenuated strains of Vibrio anguillarum. AB - The fish pathogen Vibrio anguillarum causes a lethal infection in rainbow trout (Salmo gairdneri). Three different avirulent mutants, constructed by transposon insertion mutagenesis (VAN20 and VAN70) or as antibiotic-resistant mutants (VAN1000), were isolated by screening 200 individual isolated mutants for avirulence. When used as live vaccines, all three avirulent mutants were able to induce protective immunity against the homologous as well as a heterologous strain of V. anguillarum. When VAN1000 was used, protective immunity could be recorded 1 week after bath vaccination with 10(7) bacteria per ml of water for 30 min. A single-dose immunization was effective for at least 12 weeks. Western immunoblotting showed that strains of V. anguillarum have antigenic determinants in common with Aeromonas strains. Therefore, we tested and confirmed that VAN1000 also was able to induce protective immunity against challenge with Aeromonas salmonicida. PMID- 2548447 TI - Primary endobronchial granular cell myoblastoma. AB - Granular cell myoblastoma is a common lesion of uncertain histogenesis. It commonly affects the tongue, breast, and subcutaneous tissues. However, its occurrence in the tracheobronchial tree is rare. Although generally a benign lesion, isolated malignant granular cell myoblastoma as well as its coexistence with other primary bronchogenic carcinomas has been documented. In spite of recent anecdotal reports advocating endoscopic removal of this lesion, we believe definitive surgical excision is a more rational choice of treatment. PMID- 2548448 TI - Inhibition of platelet aggregation by idebenone and the mechanism of the inhibition. AB - The inhibitory effect of 6-(10-hydroxydecyl)-2,3-dimethoxy-5-methyl-1,4 benzoquinone (idebenone) on platelet aggregation was studied in rat and human platelets in vitro, and the mechanism of inhibition was examined in rat platelets. Idebenone inhibited the aggregation induced by collagen and thrombin in washed platelets, and by arachidonate and ADP in platelet-rich plasma (PRP). The inhibition was more prominent in collagen- and arachidonate-induced aggregation. In collagen-induced aggregation of human platelets, idebenone was 8 fold more potent than aspirin. In addition, idebenone inhibited prostaglandin synthesis and thromboxane B2 production, and also increased the cyclic AMP content in platelets. However, the concentration of idebenone required to inhibit thromboxane B2 production was much lower than that required to increase cyclic AMP. These results indicate that idebenone inhibits platelet aggregation by inhibiting thromboxane B2 synthesis rather than by increasing cyclic AMP content. PMID- 2548449 TI - [Reliability of the PEHPS culture medium for the in vitro evaluation of antiamebic drugs]. AB - Monophasic liquid media for axenical cultivation of Entamoeba histolytica, have permitted to define diverse aspects of this parasite's biology, without associated organisms interference. In addition, these media have allowed the in vitro evaluation of new drugs with potential antiamebic properties. PEHPS, a new medium recently developed in our laboratory, was used in order to determine its usefulness in the searching for new antiamebic compounds. The antiamebic potency of currently employed drugs in clinical therapy: emetine, metronidazole and diiodohydroxyquinoline on E. histolytica, strain HM-1, trophozoites grown in PEHPS was tested. The IC50 for each drug was 0.082 microgram/ml, 0.10 micrograms/ml and 5.84 micrograms/ml respectively. Results obtained with each drug are statistically reliable and reproducible, and agree with the species sensibility reported by several other authors. Accordingly, we propose PEHPS as a useful medium for new antiamebic drug research. PMID- 2548450 TI - Experimental neuropathy produced in rats with industrial solvents (thinner). AB - Thinner is a very potent neurotoxic compound which damages both the central and peripheric nervous system, as well as the rest of the organism, when it is used or inhaled spontaneously chronically. 20 Wistar rats were used. They inhaled 1.5 ml of thinner per day and were sacrificed by pairs upon days 7, 15, 30, 45, 60, 75, and 90. Four of them were sacrificed after suspending the inhalation 15, 30, 60 and 90 days later so as to study the process of recuperation. The hind legs were fixated in 10% formaldehyde for 15 days. Fragments of sciatic and crural nerves were included in paraffin and histological cuts were performed both in transversal and longitudinal sections of 5 to 7 micra. Techniques for myelin and argento-auric ones were performed. Incipient lesions are described as well as intermediate and terminal ones produced after having inhaled 45 ml of thinner for 30 days. The myelin sheath shows edema, ballooning and destruction due to lysis after inhaling 135 ml in 90 days. In a parallel manner the nerve fibers show edema, irregular thickening and after inhaling 135 ml in 90 days there is atrophy or disintegration. A partial regeneration is produced if inhalation is suspended after 30 days and 30 more days are allowed for recuperation, when some Schwann cells are still active. We believe that the lesions which begin to appear on the myelin sheath occur because thinner is a fat solvent, fats being important elements in their structure and the nerve fiber injury may be secondary to the demyelinization, or due to destruction of the central neuron from which they originate. PMID- 2548451 TI - Determination of the dry matter disappearance and the crude protein degradability of the formaldehyde treated feeds in the rumen by in situ method. AB - Sunflower cake (SC), horse bean (HB), and alfalfa meal (AM) were treated with rising doses of formaldehyde (F) from 0.0, 0.2 ... 2.0 g F per 100 g crude protein. We have followed the differences of crude protein solubility, its degradability and disappearance of dry matter in the rumen as well as the amount of reversible bonded F from the F dose. With AM it was found a significant influence of ADF on the degradability and on the amount of acid-labile bonded F and/or on the decrease of N solubility. From the total N content the share of soluble nitrogen decreased in SC by 31%, HB by 24%, and in AM only by 12.5%. With the increase of the F dose its protective effect on proteins against the degradability mainly in SC and HB also rose significantly. The degradability of AM protein was influenced more by the incubation time than by the effect of treatment. The degradability of the SC proteins fell from 95% with untreated to 27% treated with the maximum dose. With AM it fell slowly from 84% to 74% by the determination coefficient R2 = 0.77. It was observed a marked decrease in HB only from the dose of 1.28% F. The changes in dry matter disappearance were similar to the changes in crude protein degradability and to the changes of solubility in SC and AM. The in situ method proved very suitable for the determination of the extent of degradability of the feed proteins in the rumen. PMID- 2548452 TI - [Studies on nitrogen metabolism in the large intestine of ruminants. 5. Metabolism of intra-cecally infused 15N-urea without and with fermentable material in heifers]. AB - Six heifers with a live weight of 215, 227 and 238 kg (experiment 1) and 220, 227 and 233 kg, resp. (experiment 2), were supplied with ileocaecal re-entrance cannulae, jugular venous catheters and bladder catheters. The ration consisted of 4 kg maize silage and 4 kg wheat straw pellets per animal per day. Up to 3.5 kg of the straw pellets, consisting of 73% wheat straw, 10% barley, 12% molasses, NPN salts and a mineral mixture, were consumed per animal per day. In a preliminary period 50% of the digesta flow was collected over 12 h/d on 5 consecutive days and stored in a deep-freeze. During the main trial the re entrance cannula was disrupted and the flowing digesta were quantitatively collected at the end of the ileum; previously collected digesta were supplemented with 15N urea and every hour over 24 h infused into the caecal part of the re entrance cannula. Between the 24th and 30th hours the digesta were infused without 15N urea supplement. In trial 2 the digesta were also supplemented with partly hydrolysed straw meal between the 1st and 30th hours (approximately 10% straw meal DM related to digesta DM). There were no differences between trials 1 and 2 with regard to the increase of atom-% 15N excess (15N') in the plasma urea. The 15N labelling decrease of the plasma urea N shows that the half-life is 7.9 h in trial 1 and 7.0 h in trial 2. The NH3 nitrogen in faeces was distinctly higher labelled in trial 2 after the supplement of straw meal than in trial 1. The total N in faeces was also twice as highly labelled as in trial 1. Atom-% 15N' in urine was significantly higher in trial 2 than in trial 1 between the 6th and 16th hours after the beginning of 15N urea supplementation. In the decrease curve of atom-% 15N' (after the 26th hour of trial) the values in trial 1 were generally higher than in trial 2. The higher bacterial protein synthesis in the large intestine in trial 2 (after the supplement of partly hydrolysed straw meal) had the effect that 13.6% of the supplemented 15N' was excreted in faeces by the 30th hour of trial, in contrast to this only 4.7% in group 1. Up to the 4th day after the 15N urea infusion these values increased to 16.2 and 6.1%, resp., only. PMID- 2548453 TI - Effect of the presence of a urea fertilizer plant on the nitrate content of berseem and constituents of milk and blood of buffaloes. AB - An attempt was made to study the effect of the presence of "El-Nasr" Fertilizer Plant in "Talkha" on the nitrate content of berseem as well as on some blood and milk constituents of buffaloes raised in the surrounding areas of the factory. The studied areas included the northern area (0-2 km from the factory), the south eastern area (1.5-3 km from the factory) and the control area (not nearer than 5 km from the factory). The study showed that the nitrate content in berseem grown near the factory had higher NO3 values exceeding 2% NO3 in DM in some cases. Berseem from the areas far from the factory had lower NO3 levels (about 50-200 ppm NO3 in DM). Samples from "El-Mansoura" and "El-Manzala" contained higher NO3 levels than the samples from "El-Senblawin" and "Belkas." There were no clear effects of cut sequence on nitrate levels in berseem. Values of blood packed cell volume and methaemoglobin percentages and plasma urea concentration did not differ significantly among the three areas (northern, south eastern and control areas). Blood haemoglobin and total protein contents were lower, whereas transaminase activity and blood nitrate contents were higher in buffaloes of the south eastern area than those for animals of the other two areas. The blood ammonia content in the control buffaloes was higher than that of the animals of the northern and south eastern areas. The analysis of milk revealed that nitrate levels were not different among areas or between morning and evening milkings. Total milk protein was higher in the northern area.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548455 TI - Restriction analysis of PBS 1-related phages. AB - The Bacillus subtilis bacteriophages AR 9, 3 NT, and I 10 belong to the same group as the widely used bacteriophage PBS 1. These four phages have in common, among other properties, flagellotropic adsorption to host cells and total substitution of thymine by uracil in their DNAs. We tested the action of different restriction endonucleases on the four genomes. Among 14 enzymes, which did cut the DNAs into discrete fragments, six are known as having at least one thymine residue in their recognition sequences. The electrophoretical patterns of the digested DNAs from the four phages show strong similarities. Nevertheless, the differences are sufficient to allow a clear distinction between them. PMID- 2548454 TI - Flavivirus infection: essential ultrastructural changes and association of Kunjin virus NS3 protein with microtubules. AB - Virus-induced vesicles evolved early in the Kunjin virus replication cycle around 9 to 10 h p.i. just before the end of the latent period in infected Vero cells. About 2 h following the appearance of the vesicles, microtubule paracrystals were also formed. These two virus-induced structures seemed interlinked and have essential roles in Kunjin virus replication. A viral protein NS3 was found to be associated with the microtubule component of the cells. When vinblastine sulphate was added to the cells immediately after infection, formation of the paracrystals was delayed by two hours, and the affiliation of NS3 protein was also observed to be rearranged. PMID- 2548456 TI - Characterization of a canine rotavirus strain by neutralization and molecular hybridization assays. AB - A canine rotavirus, RS-15, previously isolated in Japan was determined to be subgroup I and serotype 3. When compared with prototype human and animal rotavirus strains by RNA-RNA hybridization assay, the RS-15 strain showed a high degree of homology only with the canine CU-1 strain isolated in the United States of America, suggesting that canine rotaviruses constitute a distinct gene family, which we have elsewhere proposed to term "genogroup". PMID- 2548457 TI - Origin of the hemagglutinin on A/Equine/Johannesburg/86 (H3N8): the first known equine influenza outbreak in South Africa. AB - A severe influenza outbreak occurred in horses in South Africa in 1986. The causative agent was identified as an influenza virus [A/Equine/Johannesburg/86 (H3N8)]. Antigenic analyses of the hemagglutinin (HA) with ferret antisera and monoclonal antibodies showed that the Eq/Johannesburg/86 virus is similar to recent equine H3 viruses. The nucleotide sequence analysis on the HA genes of Eq/Johannesburg/86 and other equine H3 influenza viruses, together with the epidemiological data, clearly demonstrated that the Eq/Johannesburg/86 virus was derived from a virus that had been circulating in horses in the United States in 1986-87. The epidemiological information suggests that the unusually severe influenza outbreak in South Africa may be due to the lack of immunity to these viruses in the horse population. PMID- 2548458 TI - Epidemiology of rotavirus strains infecting children throughout Australia during 1986-1987. A study of serotype and RNA electropherotype. AB - The epidemiology of human rotaviruses throughout Australia was studied by examining 344 rotavirus positive faecal specimens using an enzyme immunoassay incorporating serotype specific monoclonal antibodies. Specimens were collected from children less than 5 years old admitted to urban hospitals for treatment of acute diarrhoea during the winter months of 1986 and/or 1987 in Queensland, New South Wales, Victoria, Tasmania, South Australia, and Western Australia. The infecting rotavirus serotype was identified in 229 of 344 (66.6%) specimens. The predominant serotype throughout Australia was serotype 1 which was identified in 218 of 229 (95%) typable specimens. The majority (201 of 218) were identified as monotype 1a strains. Serotype 2 strains were found in Perth, Western Australia in 2 of 12 specimens collected in 1986 and in 6 of 32 specimens collected in 1987. RNA electropherotypes comprising 30 different patterns were detected after co electrophoresis of 143 of the 218 serotyped strains. Twenty-nine electropherotypes were serologically homogeneous. One electropherotype contained strains with different monotypes including 1a, 1b, and 1d. The results show remarkable serological uniformity, associated with genetic diversity of rotavirus strains identified in widely separated areas of Australia during one winter. PMID- 2548459 TI - The inflammatory macrophage response to murine cytomegalovirus in genetically susceptible mice. AB - Adherent suppressor cells have often been implicated in the depression of immunocompetence following CMV infections. We have reported that high levels of cytostatic macrophages in the peritoneal cavities of infected mice correlate with genetically-based sensitivity to CMV disease, suggesting they may modulate protective immune responses. This study investigates the properties and kinetics of such cells. Genetically-susceptible BALB/c mice infected with MCMV accumulated activated peritoneal macrophages, 7 days post-infection. These cells suppressed 3H-thymidine-incorporation and lymphokine production in syngeneic lymphocyte cultures and hence appeared to have depressed accessory cell function, although interleukin-1 production and the capacity to take up colloidal gold were enhanced. The cytostatic activity was located in a low density fraction (1.05 g/ml), which was expanded by MCMV infection. The lowest density cells had higher frequencies of infection but the proportion of cells releasing virus (less than 0.2%) was below the proportion activated, as shown by the shift in the density profile or enhanced colloidal gold uptake. A comparable accumulation of cytostatic activated peritoneal macrophages occurred in mice treated with cyclosporine A, but nude mice showed macrophage activation without cytostasis, so the role of T cells is not resolved. The spleens of infected mice maintaining high levels of virus in this organ atrophied, and the remaining cells were unable to proliferate in culture. In contrast, mice clearing the virus developed splenomegaly and restricted responsiveness, which may be governed by cytostatic cells equivalent to those in the peritoneal cavity. The spread of virus to the lymph nodes was limited and MCMV-primed cells were readily demonstrable. PMID- 2548460 TI - Lack of quantitative correlation between inhibition of replication of rhinoviruses by an antiviral drug and their stabilization. AB - R 61,837, a new antirhinovirus compound, was able to protect several susceptible rhinoviruses against inactivation by mild acidification or heat. This observation strengthens the hypothesis that the drug exerts antiviral activity by a direct interaction with the viral protein capsid to stabilize the particle. However, the minimal concentrations necessary to inhibit either acetate or citrate or heat inactivation were different for each of five tested serotypes and we therefore conclude that stabilization and inhibition of replication are not causally linked but parallel events, both independently resulting from the binding of the drug to the viral capsid. Studies using drug resistant mutants of HRV51 and HRV9 confirmed this lack of quantitative correlation. The mutants were also shown to be cross resistant to a panel of seven different reference antirhinoviral drugs including SDS, WIN51711, chalcone, dichloroflavan and MDL20,610. This indicates that all these compounds bind to the same site corresponding to the hydrophobic pocket within the viral protein VP 1 beta-barrel structure of HRV14. PMID- 2548461 TI - Molecular epidemiology and restriction site mapping of adenovirus 7 genome types. AB - From the United States, the Federal Republic of Germany, and other regions, 168 strains of AV7, isolated between 1961 and 1985, were analyzed by six restriction endonucleases and nine genome types were identified. The enzymes BamHI and HindIII were most discriminative. The genome type D5 (or 7b) predominated with 120 isolates since 1970 in both countries. Strains of D2 (7a) and D4 (7c) were isolated for a limited time, D3 for an extended time period. Several clusters of infections with the same genome type were found. Differences in pathogenicity could not be derived from our data. On the basis of restriction site mapping, most other genome types were similar to D5, one to D2 and one to the prototype (D1). The genomic relation between AV7 and AV3 is discussed and shown by a dendrogram. PMID- 2548462 TI - Genetic and antigenic differences between fowlpox and quailpox viruses. AB - The genomes of a fowlpox and quailpox virus isolate were compared by restriction enzyme analysis using BamHI, EcoRI, and HindIII endonucleases. The genetic profiles of the two virus species were very distinct with fragments lacking similar electrophoretic mobilities. In contrast, the patterns of three quailpox virus isolates were very similar with a high proportion of co-migrating fragments. When the immunogenic proteins of two quailpox, three fowlpox, a juncopox, and a pigeonpox virus isolate were examined by immunoblotting, common as well as unique antigens were detected. The greatest disparity was between quailpox virus and the other three species which are genomically conserved. Therefore, on the basis of genetic as well as immunological analysis, quailpox virus is a distinct species of the genus Avipoxvirus. PMID- 2548464 TI - [Glomus tumors of the temporal bone: a report of 6 cases]. AB - The authors review the clinical, radiological and pathological features of 6 cases of glomus tumors of the temporal bone. Out of the 6 patients, 5 were female; age was distributed between 22 and 76 years (mean 48 years). The main clinical features were hypoacusia, tinnitus and otoscopic findings suggestive of the diagnosis. In one case was noted the concomitant presence of a neurinoma of the VIII cranial nerve with a ipsilateral glomus tumor, and in another case there was a concomitancy of carotid body tumor with temporal glomus jugularis tumor. Metastases were not observed in any case. Tumoral lesions were successfully ressected employing microsurgical techniques and a multidisciplinary staff involving neurosurgeons, head and neck surgeons and otolaryngologists. Radioteraphy was not employed, neither pre-operative embolization. Some aspects related to the nosology, embriology, pathophysiology, diagnosis and treatment of this interesting type of neoplasms are discussed. PMID- 2548463 TI - Characterization of a non-occluded baculovirus (subgroup C) from the field cricket, Gryllus rubens. AB - A non-occluded baculovirus was isolated from nymphs of the field cricket, Gryllus rubens. SDS-polyacrylamide gel electrophoresis revealed the presence of 6 major and 11 minor polypeptides in these particles. Restriction endonuclease analysis indicated that the genome, 87.0 +/- 1.8 kilobase pairs, was a closed circular DNA molecule. DNA-DNA hybridization in low strigency conditions revealed no homology with the genomes of Oryctes baculovirus or Autographa california NPV. The virus replicated in nuclei of fat body cells, and was transmitted per os to a small proportion of first instar G. rubens nymphs. PMID- 2548465 TI - Erythrocyte sodium-potassium pump in thyrotoxic periodic paralysis. AB - To search for a genetic marker in patients with thyrotoxic periodic paralysis (TPP), we studied the erythrocyte sodium-potassium pump activity in 13 patients with TPP; 30 thyrotoxic patients with no history of paralysis (T) and 69 euthyroid controls. In thyrotoxic patients (TPP and T), erythrocyte ouabain binding and ouabain sensitive sodium efflux rate constant were decreased while erythrocyte sodium content was increased. All these changes reverted to normal when the patients became euthyroid. Maximal ouabain binding capacity correlated positively with ouabain sensitive sodium efflux rate constant (r = 0.542; p less than 0.001; n = 155) and negatively with serum thyroxine concentration (r = 0.571; p less than 0.001; n = 60) and erythrocyte sodium content (r = -0.521; p less than 0.001; n = 155). In the thyrotoxic state, maximal ouabain binding capacity was just significantly higher in TPP when compared with T (0.268 +/- 0.014 and 0.234 +/- 0.009 pmol/10(9) cells respectively; p less than 0.05). This difference could not be demonstrated when the patients became euthyroid. Our findings suggest that patients with TPP respond to thyrotoxicosis with a smaller decrement in erythrocyte sodium-potassium ATPase activity than patients without a history of paralysis. However, the difference is too small to represent a useful genetic marker for this disease entity PMID- 2548466 TI - Hepatic resection: value of operative ultrasound and ultrasonic dissection. AB - Twenty-five patients having a major liver resection at the Austin Hospital were reviewed, 18 of whom had their operation after 1986. The indications were for metastatic disease in 14, hydatid in six and primary tumour in five. There were no operative deaths. Dissection was performed with the Cavitron Ultrasound Surgical Aspirator (CUSA) and peroperative ultrasound used in 18 and without in seven. Mean blood loss was 1.75 u in the CUSA group and 6.5 u in the non-CUSA group (P less than 0.05). It is concluded that intra-operative ultrasound and use of the CUSA makes major liver resection a more precise and less hazardous procedure. PMID- 2548468 TI - Multicentric chemodectoma in the head and neck. AB - A case of a multicentric chemodectoma, with the unusual combination of glomus vagale and glomus tympanicum, is presented. The patient, though asymptomatic, showed elevated levels of urinary catecholamines, suggesting some biochemical activity of the tumour. Multicentricity was unsuspected prior to selective carotid arteriography. Intra-operative haemostasis was assisted by the utilization of pre-operative selective embolization of the tumour mass. Subsequently, the injected Ivalon was confirmed histologically in the specimen. The asymptomatic small glomus tympanicum tumour was treated using embolization alone but only time will determine the effectiveness of this method of management. The vagus nerve was anatomically spared, but complete function had not returned after 2 years of follow-up. PMID- 2548467 TI - Biliary obstruction due to intraductal tumor. AB - Five unusual cases of bile-duct obstruction are presented. There were three cases of intrabiliary hepatoma, one of intrabiliary colon cancer metastasis, and one cystadenoma of the bile-duct. The importance of an accurate diagnosis is emphasized, as these tumours can be treated by surgery or by biliary stenting, sometimes with prolonged survival. PMID- 2548469 TI - Morbidity from acquired cytomegalovirus infection in a neonatal intensive care unit. AB - In a prospective study of transfused neonates, 32 of 262 infants were viruric at greater than 20 days of life. Of 212 neonates whose early status was known, postnatally acquired infection was proven in 21, two of whom were seronegative at birth and were thought to have transfusion-acquired cytomegalovirus (CMV). Maternal transmission of CMV is important in this population as there was 91% seropositivity for CMV at birth among the 21 babies who acquired CMV compared with 55% positivity among 150 babies who did not shed CMV (P less than 0.01). Significantly increased morbidity (increased length of stay in hospital, increased use of antibiotics, and longer duration of antibiotic administration) was found in babies with acquired CMV compared with matched controls who did not become viruric. Significant morbidity and mortality was not restricted to the two seronegative babies with transfusion-acquired CMV. The cost of providing CMV antibody negative blood for this neonatal unit would be less than the cost of providing the extended hospital care needed by the two babies with transfusion acquired CMV found during this 3 year study. PMID- 2548470 TI - Sensitivity to oxidants of mitochondrial and sarcoplasmic reticular calcium uptake in saponin-treated cardiac myocytes. AB - Calcium transport functions of mitochondria and sarcoplasmic reticulum (SR) were studied without prior extraction using isolated rat heart myocytes permeabilized with saponin. Calcium uptake by SR was rapid and its affinity was high in comparison to calcium uptake by mitochondria, which had a higher capacity. The sensitivity of uptake to two oxidants, H2O2 and HOCl (hypochlorous acid), depended on the cytosolic calcium concentration; when this was similar to the concentration in diastole (180 nM), HOCl inhibited calcium uptake by mitochondria and SR, whereas when the calcium concentration was 750 nM, mitochondrial calcium uptake showed relatively high resistance, although SR uptake was still markedly inhibited by HOCl. Calcium uptake of both mitochondria and SR was less sensitive to the action of H2O2 than to HOCl, and the H2O2 effect was less dependent on the cytosolic calcium concentration. Therefore, HOCl, when produced by activated leukocytes and supplied to the heart cells, may seriously impair the excitation contraction coupling function of SR, whereas H2O2, possibly generated directly by mitochondria or generated from superoxide anions, may be tolerated relatively well by heart SR and mitochondria. PMID- 2548471 TI - Effect of ouabain on hemodynamics in acute volume expanded hypertensive dogs. AB - To evaluate possible roles of endogenous Na+-K+-ATPase inhibitors in vasoconstricted blood pressure elevation produced by acute volume expansion, we administered ouabain (Na+-K+-ATPase inhibitor) intravenously (30 micrograms/kg) for 10 min to dogs, 3 h after volume expansion with dextran in lactated Ringer's solution (20 ml/kg, for 1 h). Acute volume expansion resulted in the elevation of blood pressure associated with an increase in cardiac output. In some dogs the blood pressure remained elevated with gradual increase in total peripheral resistance (Group I) or with sustained high cardiac output (Group II), and in other dogs (Group III) it returned to the control level. Ouabain administration elevated the blood pressure and total peripheral resistance in these groups and sham dogs which did not have volume expansion. And these effects of ouabain were not correlated with the degree of blood pressure or vasoconstriction produced by volume expansion. Thus, it is not likely that endogenous Na+-K+-ATPase inhibitors increased to produce vasoconstricted hypertension after acute volume expansion. PMID- 2548472 TI - [Results of surgical treatment of syndactylia of the hand]. AB - The Syndaktyly is one of the most frequent failures of the hand. The aim of its operative treatment is a functional improvement of the developing hand and its aesthetics. The right moment and method of operation are decisive for the functional and aesthetics success as the kinds of Syndaktyly vary. Between 1970 and 1985 96 children have been operated on the hand because of Syndaktyly and its varieties at the Halle-Surgical-University-Hospital. Experiences and results during the treatments of this misdevelopment are being reported in a study of postoperative treatment over period of 15 years. PMID- 2548473 TI - [Testing the effectiveness of foot-and-mouth disease vaccines: the relationship between test infection results and corresponding neutralization titers of vaccinated cattle]. AB - In the course of vaccine controls, the potency of 25 foot-and-mouth disease (FMD) vaccines was tested quantitatively in parallel in cattle using the intradermal infection and the determination of the SN titres. More than 95% of the vaccinated cattle with SN titres of greater than 1:20 were protected from generalized FMD, regardless of the virus type tested. 61.5% of the vaccinated cattle with SN titres less than or equal to 1:20 were not protected and developed generalized FMD. Comparison of the PD50 values calculated from the results of the intradermal infection and the corresponding SN titres (minimum protection titre greater than 1:20) showed that the results were in complete agreement in 56% of the tested vaccines. In a further 32% of vaccines, the PD50 calculated from the SN titre was slightly below that for the intradermal infection, in the remaining 12% it was somewhat above. The possibility of using the minimum titre determination for testing a vaccine and the significance of this titre as an expression of protection by vaccination are discussed. PMID- 2548475 TI - Evidence for intracellular sodium pumps in permeabilized Xenopus laevis oocytes. AB - Ouabain binding was studied in Xenopus laevis oocytes permeabilized by detergents. The behaviour of markers showed that 10 microM-digitonin selectively disrupts the plasma membrane. In the presence of ATP, oocytes permeabilized at 10 microM-digitonin bound no more ouabain molecules than were required to abolish active 86Rb+ uptake in the intact cells. However, the ouabain binding capacity increased approx. 2-fold when inner membranes were disrupted by SDS or excess digitonin, as judged from the accompanying release of the lysosomal marker beta hexosaminidase. The results suggest that oocytes have a large internal pool of functional sodium pumps. PMID- 2548474 TI - Metabolism of the inositol phosphates produced upon receptor activation. PMID- 2548476 TI - Folate (pteroylglutamate) uptake in human red blood cells, erythroid precursors and KB cells at high extracellular folate concentrations. Evidence against a role for specific folate-binding and transport proteins. AB - Membrane-associated folate (pteroylglutamate, PteGlu)-binding proteins (FBPs) play an important role as PteGlu-transport proteins in malignant and normal human cells. Since high extracellular folate (PteGlu) concentrations (EFC) profoundly influenced uptake and toxicity of the anti-PteGlu methotrexate in malignant KB cells, we studied human cells to determine additional mechanisms for PteGlu uptake when the EFC was varied. At low EFC (less than 10 nM), the predominant mechanism for folate uptake in mature erythrocytes was through binding to externally oriented FBPs which were quantitatively insignificant (4-6 orders of magnitude lower) and of no apparent physiological relevance when compared with KB cells. However, the predominant mechanism of PteGlu accumulation at high EFC [10 250 nM] in intact erythrocytes and sealed right-side-out (RSO) ghosts was not FBP mediated and non-specific. This conclusion was based on the findings that radiolabelled PteGlu uptake: (i) continued even in the presence of a 1000-fold excess of unlabelled PteGlu and was linear and not saturable up to 250 nM; (ii) was two-fold higher at pH 4.5 than 7.5; (iii) was less than 2-fold increased at 37 degrees C compared with 4 degrees C; and (iv) was unaffected after trypsin mediated proteolysis of greater than 75% FBPs. The [3H]PteGlu and 125I-PteGlu (histamine derivative) accumulated intracellularly through the non-specific PteGlu-uptake mechanism was unaltered biochemically and in a soluble compartment. Raising the EFC 500-fold higher than controls during erythropoiesis in vitro resulted in reversal of the expected anti-(placental folate-receptor)-antiserum induced megaloblastic changes in orthochromatic normoblasts derived from burst forming unit-erythroid colonies. Furthermore, at EFC greater than 0.1 microM, KB cell accumulation of [3H]PteGlu was also predominantly through a mechanism that did not involve specific FBPs. Thus, at high EFC, a major component of PteGlu transport in human cells is not mediated through FBPs and is likely to be a passive diffusion process. PMID- 2548477 TI - Evidence that activation of a common G-protein by receptors for leukotriene B4 and N-formylmethionyl-leucyl-phenylalanine in HL-60 cells occurs by different mechanisms. AB - Differentiated HL-60 cells were found to respond to the chemoattractants leukotriene B4 (LTB4) and N-formylmethionyl-leucyl-phenylalanine (FMLP), in a manner similar to neutrophils. Membranes of myeloid differentiated HL-60 cells were used (a) to examine the ability of LTB4 receptors to interact with a guanine nucleotide-binding protein (G-protein), and (b) to compare this G-protein with that which is coupled to the FMLP receptor. LTB4 stimulated a dose-dependent increase in GTP hydrolysis and guanosine 5'-[gamma-thio]triphosphate (GTP[S]) binding, demonstrating that LTB4 receptors on HL-60 cells are coupled to a G protein. Both pertussis toxin and cholera toxin inhibited stimulation of GTPase activity and GTP[S] binding by either LTB4 or FMLP, indicating that both receptors are coupled to a G-protein containing a 40 kDa alpha-subunit. That the two receptors share a common G-protein was shown by FMLP enhancement of cholera toxin-induced inhibition of GTPase activity stimulated by either FMLP or LTB4. However, LTB4 did not enhance cholera-toxin-induced inhibition of GTPase activity, suggesting that the receptors interacted differently with this G protein. This difference was confirmed by showing that FMLP, but not LTB4, stimulated receptor-specific [32P]ADP-ribosylation of the 40 kDa alpha-subunit. Concentrations of LTB4 and FMLP which produced maximal responses produced enhanced stimulation in both assays. This additive effect was not abolished by inactivation of up to 80% of G-protein activity by N-ethylmaleimide or cholera toxin. We conclude that LTB4 and FMLP receptors in HL-60 cells are coupled to a common G-protein. The receptor--G-protein interaction is different for the two receptors, and G-proteins not coupled to both receptors may account for the additive response. PMID- 2548478 TI - Quantitative determination of Ca2+-dependent Mg2+-ATPase from sarcoplasmic reticulum in muscle biopsies. AB - The possibility of quantifying the total concentration of Ca2+-dependent Mg2+ ATPase of sarcoplasmic reticulum was investigated by measurement of the Ca2+ dependent steady-state phosphorylation from [gamma-32P]ATP and the Ca2+-dependent 3-O-methylfluorescein phosphatase (3-O-MFPase) activity in crude muscle homogenates. The Ca2+-dependent phosphorylation at 0 degree C (mean +/- S.E.) was 40.0 +/- 2.5 (n = 6) and 6.2 +/- 0.7 (n = 4) nmol/g wet wt. in rat extensor digitorum longus (EDL) and soleus muscle, respectively (P less than 0.001). The Ca2+-dependent 3-O-MFPase activity at 37 degrees C was 1424 +/- 238 (n = 6) and 335 +/- 56 (n = 4) nmol/min per g wet wt. in rat EDL and soleus muscle, respectively (P less than 0.01). The molecular activity calculated from these measurements amounted to 35 +/- 5 min-1 (n = 6) and 55 +/- 10 min-1 (n = 4) for EDL and soleus muscle respectively. These values were not different from the molecular activity calculated for purified Ca2+-ATPase (36 min-1). The Ca2+ dependent 32P incorporation in soleus muscle decreased in the order mice greater than rats greater than guinea pigs. In EDL muscles from hypothyroid rats at a 30% reduction of the Ca2+-dependent phosphorylation was observed. The Ca2+-dependent phosphorylation in vastus lateralis muscle from three human subjects amounted to 4.5 +/- 0.8 nmol/g wet wt. It is concluded that measurement of the Ca2+-dependent phosphorylation allows rapid and reproducible quantification of the concentration of Ca2+-dependent Mg2+-ATPase of sarcoplasmic reticulum. Since only 20-60 mg of tissue is required for the measurements, the method can also be used for biopsies obtained in clinical studies. PMID- 2548479 TI - Effects of diethyldithiocarbamate and endogenous polyamine content on cellular responses to hydrogen peroxide cytotoxicity. AB - In exponential-phase Chinese-hamster cells, 0.1 mM-diethyldithiocarbamate (DDC) afforded greater than 1 log survival protection to cultures treated before and during exposure to 1 mM-H2O2. Both DDC and H2O2 treatment stimulated the activity of ornithine decarboxylase (ODC), the first enzyme in polyamine synthesis, within 4 h of exposure. DDC, and to a lesser degree H2O2, also stimulated the activity of spermidine N1-acetyltransferase (SAT), the rate-limiting enzyme in polyamine catabolism. The increase in SAT activity, after exposure to DDC or another stress (heat shock), was inhibited in cells depleted of putrescine and spermidine by alpha-difluoromethylornithine (DFMO), the enzyme-activated suicide inhibitor of ODC. Pretreatment with DFMO or heat shock also induced resistance to H2O2 cytotoxicity. Since SAT activity is low in resting cells, yet stimulation of enzyme activity depends on endogenous spermidine pools, these results suggest that the expression of SAT activity occurs by a mechanism involving a stress dependent displacement of spermidine into a new intracellular compartment. The stimulation of ODC and SAT activities does not appear to be a necessary component of the mechanism by which DDC protects cells from H2O2 cytotoxicity, although spermidine displacement may be a common facet of the cellular response to stress. PMID- 2548480 TI - Platelet-derived-growth-factor-stimulated heterogeneous polyphosphoinositide metabolism and phosphate uptake in C3H fibroblasts. AB - Pig platelet-derived growth factor (PDGF) increased the rate of [32P]Pi uptake by murine fibroblasts, resulting in a 3-9-fold elevation of the specific radioactivity of ATP, PtdInsP, PtdInsP2, PtdIns and phosphatidic acid. The specific radioactivity was 10-60-fold higher in ATP than in the four phospholipids. These substances are therefore not in metabolic equilibrium, which complicates determination of inositol phospholipid turnover. PMID- 2548482 TI - The role of N-linked glycosylation in the regulation of activity of 3-hydroxy-3 methylglutaryl-coenzyme A reductase and proliferation of SV40-transformed 3T3 cells. AB - The effects of glycosylation inhibitors on the proliferation of SV40-transformed 3T3 cells (SV-3T3) were examined in vitro. Whereas swainsonine and castanospermine, which inhibit distal steps in the glycosylational processing, exerted marginal or no effects on cell proliferation, a proximal inhibitor, tunicamycin, efficiently decreased the rate of DNA synthesis and also inhibited the activity of 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase. The inhibitory effects of tunicamycin on cell proliferation could be partially reversed by addition of dolichol, a metabolite in the pathway regulated by HMG CoA reductase. This finding suggests that tunicamycin exerts at least one of its effects on cell proliferation by modulating the activity of HMG-CoA reductase. PMID- 2548483 TI - A model for the initiation of translation at the late stage after infection by adenoviruses. PMID- 2548481 TI - The Ins(1,4,5)P3 binding site of bovine adrenocortical microsomes: function and regulation. AB - Adrenocortical microsomes possess a single population of Ins(1,4,5)P3-specific binding sites [IC50 5.9 +/- 0.9 nM; Palmer, Hughes, Lee & Wakelam (1988) Cell. Signalling 1, 147-156]. Competition studies showed that Ins(1:2-cyclic,4,5)P3 exhibits a 21-fold lower affinity for the site than Ins(1,4,5)P3 (IC50 124 +/- 16 nM). The affinity of the binding sites for Ins(1,4,5)P3 was not influenced by the non-hydrolysable GTP analogues GTP gamma S and Gpp[NH]p or by preincubation of the binding protein with a preparation of partially purified protein kinase C in the presence of ATP and TPA (12-O-tetradecanoylphorbol 13-acetate). These observations are discussed with reference to the identify and function of the Ins(1,4,5)P3 binding site. PMID- 2548484 TI - Influence of bacterial toxins and forskolin upon vasopressin-induced inositol phosphate accumulation in WRK 1 cells. AB - The accumulation of inositol phosphates in WRK 1 cells, stimulated with a range of vasopressin concentrations, was diminished by prior exposure to cholera toxin or forskolin, whilst that observed in the presence of maximal concentrations of the hormone was enhanced in pertussis-toxin-treated cells. In the presence of [32P]NAD+, both cholera toxin and pertussis toxin provoked the labelling of peptides with approximate Mrs of 45,000 and 41,000 respectively in the membranes of WRK 1 cells. Exposure to cholera toxin or forskolin for 15-18 h enhanced cyclic AMP accumulation in these cells. The concentrations of these agents which provoked half-maximal cyclic AMP accumulation were similar to those required to diminish receptor-mediated inositol phosphate accumulation by 50%. In contrast, half-maximal ADP-ribosylation of the 45,000Mr peptide needed 100-fold greater concentrations of the toxin than were effective in provoking half-maximal inhibition of inositol phosphate accumulation. Cholera toxin or forskolin also reduced the maximal specific binding, to intact WRK 1 cells, of both [3H][Arg8]vasopressin and the V1a antagonist [3H][beta-mercapto-beta,beta cyclopentamethylenepropionic acid,O-methyl-Tyr2, Arg8]vasopressin. The kinetics for the loss of this binding capacity following cholera-toxin treatment were very similar to those describing the diminution of vasopressin-stimulated inositol phosphate accumulation in the same cells. PMID- 2548486 TI - Overexpression and site-directed mutagenesis of the succinyl-CoA synthetase of Escherichia coli and nucleotide sequence of a gene (g30) that is adjacent to the suc operon. AB - The succinyl-CoA synthetase of Escherichia coli is encoded by two genes, sucC (beta subunit) and sucD (alpha subunit), which are distal genes in the sucABCD operon. They are expressed from the suc promoter, which also expresses the dehydrogenase and dihydrolipoyl succinyl-transferase subunits of the 2 oxoglutarate dehydrogenase complex. Strategies have now been devised for the site directed mutagenesis and independent expression of the succinyl-CoA synthetase (alpha 2 beta 2 tetramer) and the individual subunits. These involve (1) subcloning a promoterless sucCD fragment downstream of the lac promoter in M13mp10, and (2) precise splicing of the suc coding regions with the efficient atpE ribosome-binding site and expression from the thermoinducible lambda promoters in the pJLA503 vector. Succinyl-CoA synthetase specific activities were amplified 40-60-fold within 5 h of thermoinduction of the lambda promoters, and the alpha and beta subunits accounted for almost 30% of the protein in supernatant fractions of the cell-free extracts. Site-directed mutagenesis of potential CoA binding-site residues indicated that Trp-43 beta and His-50 beta are essential residues in the beta-subunit, whereas Cys-47 beta could be replaced by serine without inactivating the enzyme. No activity was detected after the histidine residue at the phosphorylation site of the alpha-subunit was replaced by aspartate (His-246 alpha----Asp), but this alteration seemed to have a deleterious effect on the accumulation of the enzyme in cell-free supernatant extracts. The nucleotide sequence of an unidentified gene (g30) that is adjacent to the sucABCD operon was defined by extending the sequence of the citric acid cycle gene cluster by 818 bp to 13379 bp: gltA-sdhCDAB-sucABCD-g30. This gene converges on the suc operon and encodes a product (P30) that contains 230 amino acids (Mr 27,251). Highly significant similarities were detected between the N terminal region of P30 and those of GENA [the product of another unidentified gene (geneA) located upstream of the aceEF-lpd operon], and GNTR (a putative transcriptional repressor of the gluconate operon of Bacillus subtilis). Possible roles for GENA and P30 as transcriptional regulators of the adjacent operons encoding the pyruvate and 2-oxoglutarate dehydrogenase complexes are discussed. PMID- 2548487 TI - Mode of activation of bovine brain inositol 1,4,5-trisphosphate 3-kinase by calmodulin and calcium. AB - The effect of Ca2+ and calmodulin (CaM) on the activation of purified bovine brain Ins(1,4,5)P3 kinase was quantified and interpreted according to the model of sequential equilibria generally used for other calmodulin-stimulated systems. Two main conclusions can be drawn. (i) CaM.Ca3 and CaM.Ca4 together are the biologically active species in vitro, as is the case for the great majority of other calmodulin targets. (ii) These species bind in a non-co-operative way to the enzyme with an affinity constant of 8.23 x 10(9) M-1, i.e. approx 10-fold higher than for most calmodulin-activated target enzymes. The dose-response curve of the activation of Ins(1,4,5)P3 kinase by calmodulin is not significantly impaired by melittin and trifluoperazine, whereas under very similar assay conditions the half-maximal activation of bovine brain cyclic AMP phosphodiesterase requires over 30-50-fold higher concentrations of CaM when 1 microM melittin or 20 microM-trifluoperazine is present in the assay medium. Similarly, 1 microM of the anti-calmodulin peptides seminalplasmin and gramicidin S, as well as 20 microM of N-(6-aminohexyl)-5-chloro-1-naphthalene-sulphonamide (W7), do not inhibit the activation process. These data suggest that binding and activation of Ins(1,4,5)P3 kinase require surface sites of calmodulin which are different from those involved in the binding of most other target enzymes or of model peptides. PMID- 2548485 TI - Modulation of nuclear cyclic AMP-dependent protein kinase in dibutyryl cyclic AMP treated rat H4IIE hepatoma cells. AB - Biochemical and immunochemical studies were undertaken to quantify the effects of cyclic AMP on cyclic AMP-dependent protein kinase subunit levels in nuclei of H4IIE hepatoma cells. Dibutyryl cyclic AMP (10 microM) caused a significant biphasic (10 and 120 min after stimulation) increase in total nuclear protein kinase activity. The increase observed 10 min after dibutyryl cyclic AMP stimulation was primarily due to an approx. 3-fold increase of catalytic (C) subunit activity, whereas the change observed 120 min after stimulation consisted of an increase in both C subunit and cyclic AMP-independent protein kinase activities. Analysis of nuclear protein extracts by photoaffinity labelling with 8-azido cyclic [32P]AMP identified only the type II regulatory subunit (RII), but not the type I regulatory subunit (RI). Analysis of nuclear RII variants by two dimensional gel electrophoresis demonstrated that dibutyryl cyclic AMP caused the appearance of two RII variant forms which were not present in the nuclei of unstimulated cells. Using affinity-purified polyclonal antibodies and immunoblotting procedures, we identified an approx. 2-fold increase in the RII and C subunits in nuclear extracts of dibutyryl cyclic AMP-treated hepatoma cells. Finally, the RI, RII and C subunits were quantified by an e.l.i.s.a. which indicated that dibutyryl cyclic AMP increased nuclear RII and C subunits levels biphasically, reaching peak values 10 and 120 min after the initial stimulation. Nuclear RI subunit levels were not affected. These results provide qualitative as well as quantitative evidence for a modulation by cyclic AMP of the nuclear RII and C subunit levels in rat H4IIE hepatoma cells, and indicate a relatively rapid but temporarily limited dibutyryl cyclic AMP-induced translocation of the RII and C subunits to nuclear sites. PMID- 2548488 TI - Vasopressin-stimulated Ca2+ influx in rat hepatocytes is inhibited in high-K+ medium. AB - We examined the effects of K+ substitution for Na+ on the response of hepatocytes to vasopressin, and on the hepatocyte plasma-membrane potential. (1) High K+ (114 mM) had no effect on the initial increase in phosphorylase a activity in response to vasopressin, but abolished the ability of the hormone to maintain increased activity beyond 10 min. With increasing concentrations a decrease in the vasopressin response was first observed at 30-50 mM-K+. (2) High K+ (114 mM) had no effect on basal 45Ca2+ influx, but abolished the ability of vasopressin to stimulate influx. This effect was also first observed at a concentration of 30-50 mM-K+. (3) Increasing K+ had little effect on the plasma-membrane potential until a concentration of 40 mM was reached. With further increases in concentration the plasma membrane was progressively depolarized. (4) Replacement of Na+ with N methyl-D-glucamine+ depolarized the plasma membrane to a much smaller extent than did replacement with K+, and was also much less effective in inhibiting the vasopressin response. (5) The plasma-membrane potential was restored to near the control value by resuspending cells in normal-K+ medium after exposure to high-K+ medium. The effects of vasopressin on phosphorylase activity were also restored. (6) We conclude that the Ca2+ channels responsible for vasopressin-stimulated Ca2+ influx are closed by depolarization of the plasma membrane. PMID- 2548489 TI - Discovery of a protein phosphatase activity encoded in the genome of bacteriophage lambda. Probable identity with open reading frame 221. AB - Infection of Escherichia coli with phage lambda gt10 resulted in the appearance of a protein phosphatase with activity towards 32P-labelled casein. Activity reached a maximum near the point of cell lysis and declined thereafter. The phosphatase was stimulated 30-fold by Mn2+, while Mg2+ and Ca2+ were much less effective. Activity was unaffected by inhibitors 1 and 2, okadaic acid, calmodulin and trifluoperazine, distinguishing it from the major serine/threonine specific protein phosphatases of eukaryotic cells. The lambda phosphatase was also capable of dephosphorylating other substrates in the presence of Mn2+, although activity towards 32P-labelled phosphorylase was 10-fold lower, and activity towards phosphorylase kinase and glycogen synthase 25 50-fold lower than with casein. No casein phosphatase activity was present in either uninfected cells, or in E. coli infected with phage lambda gt11. Since lambda gt11 lacks part of the open reading frame (orf) 221, previously shown to encode a protein with sequence similarity to protein phosphatase-1 and protein phosphatase-2A of mammalian cells [Cohen, Collins, Coulson, Berndt & da Cruz e Silva (1988) Gene 69, 131-134], the results indicate that ORF221 is the protein phosphatase detected in cells infected with lambda gt10. Comparison of the sequence of ORF221 with other mammalian protein phosphatases defines three highly conserved regions which are likely to be essential for function. The first of these is deleted in lambda gt11. PMID- 2548491 TI - Demonstration of receptors for parathyroid hormone on cultured rabbit costal chondrocytes. AB - Parathyroid hormone (PTH) receptors on cultured rabbit costal chondrocytes were demonstrated using HPLC-purified, radioiodinated [Nle8,-Nle18, Tyr34] bovine PTH (1-34)amide. PTH binding was found to be specific for PTH agonists and antagonists and dependent on the time and temperature of incubation. Both growth cartilage (GC) cells and resting cartilage (RC) cells were shown to have a single class of saturable, high affinity PTH binding sites with a dissociation constant of 0.6-0.7 nM. However, the numbers of receptors per cell were approximately 49,000 on GC cells and 19,000 on RC cells. After crosslinking the receptors on these cells with the radioligand, one, major 125I-labeled band of 76 kDa was separated by SDS-PAGE. PMID- 2548490 TI - Transcriptional inhibitors affecting topoisomerase II induce changes in histone methylation patterns similar to those induced by heat shock. AB - The relationship between gene expression and the patterns of histone methylation in Drosophila cells has been investigated using inhibitors of transcription acting at various levels. Inhibition of ribosomal RNA synthesis and processing by 5-fluorouridine or of general RNA synthesis by camptothecin, an inhibitor of topoisomerase I, does not affect the methylation pattern of core histones. This suggests that the arrest of transcription per se is not involved in the changes in histone methylation such as those encountered in heat-shocked cells. However, ethidium bromide and novobiocin, which are known to disrupt nucleosome structure, and VM-26 (teniposide), a specific inhibitor of topoisomerase II, induce changes in histone methylation patterns which, though less severe, are similar to those observed under cellular stress. These results suggest that chromatin conformation is probably an important factor in the accessibility of histones to methyltransferases. PMID- 2548492 TI - Calcium influx mediated by nicotinic receptors and voltage sensitive calcium channels in SK-N-SH human neuroblastoma cells. AB - When SK-N-SH human neuroblastoma cells were exposed to nicotine (NIC) or KCl they showed a dose-dependent transient increase (2- to 4-fold) in intracellular Ca2+ concentration ([Ca2+])i as detected by quin-2 fluorescence, with half maximal effects (EC50) observed at 13 microM and 26 mM, respectively. Tubocurarine and 1 isodihydrohistrionicotoxin potently blocked the NIC-evoked (IC50 congruent to 1 microM and 0.3 microM, respectively), but not the high [K+]o-evoked [Ca2+]i accumulation. The KCl-induced response was inhibited by verapamil and diltiazem (IC50 = 1.4 and 10.9 microM, respectively). Tetrodotoxin (3 microM) and tetraethylammonium (10 microM) had no effect on [Ca2+]i accumulation induced by either agent. Increases in [Ca2+]i could be evoked sequentially by NIC and KCl in the same cells suggesting independent mechanisms of Ca2+ entry. In a Ca2+-free medium, no response to either KCl or NIC was observed. However, when Ca2+ ions were restored, [Ca2+]i accumulation was enhanced to the same extent as cells suspended in a Ca2+-containing buffer. Long-term (18 hr) pretreatment of SK-N-SH cells with pertussis (100 ng/ml) or cholera toxins (10 nM) had no effect on NIC or KCl-induced [Ca2+]i accumulation. Together, these data demonstrate the presence of NIC receptors and voltage-sensitive Ca2+ channels on SK-N-SH neuroblastoma cells, through which [Ca2+]i may be modulated. PMID- 2548493 TI - Elevation of c-myc transcript level in human liver during surgical resection of hepatocellular carcinoma: possible cause for underestimation of c-myc gene activation in the tumor. AB - It has been a matter of controversy as to whether c-myc gene expression is activated in human hepatocellular carcinoma (HCC). We observed that the c-myc mRNA level in HCC was similar to that of the adjacent non-cancerous portion, as determined in freshly obtained specimens after a partial liver resection. However, the c-myc transcript was at a low level in non-cancerous tissue which was biopsied prior to surgery, whereas it was still at a high level in HCC obtained without performing hepatectomy. These results suggest that the high transcript level observed in the non-cancerous tissue from the excised liver relates to liver resection itself, and that the c-myc gene expression is enhanced in the HCC. PMID- 2548494 TI - Inositol phospholipids activate plasma membrane ATPase in plants. AB - Phosphatidylinositol-4-monophosphate and phosphatidylinositol-4,5-bisphosphate increased the activity of the vanadate-sensitive ATPase associated with plasma membranes isolated from both sunflower hypocotyls and carrot suspension culture cells. The response was not due to the metabolism of the polyphosphoinositides since diacylglycerol, inositol-1,4-bisphosphate, inositol-1,4,5-trisphosphate, glycerophosphoinositol monophosphate and glycerophosphoinositol bisphosphate had no effect. These data suggest that activation of the inositol phospholipid kinases could be a critical step in signal transduction in plants. PMID- 2548495 TI - Effect of endothelin on Ca2+ influx, intracellular free Ca2+ levels and ligand binding to N and L type Ca2+ channels in rat brain. AB - The actions of endothelin, an endogenous vasoconstrictor compound with potent effects on various parameters of Ca2+ metabolism in peripheral tissue, were studied in several neuronal preparations. Endothelin, by itself, did not alter resting intracellular free Ca2+ levels or Ca2+ influx in either rat or chicken brain preparations; nor did it affect depolarization (K+) induced changes in these parameters. Endothelin also had no effect on the binding of [3H] nitrendipine or [125I]-omega-conotoxin to "L " or "N" type channels respectively nor did it induce the release of endogenous acetylcholine from brain slices. The results show that, despite the proposed role of endothelin on voltage sensitive Ca2+ channels in peripheral tissue and despite the existence of endothelin binding sites on both smooth muscle and neurons, endothelin has no measurable effects on Ca2+ metabolism in neural tissue of central origin. PMID- 2548496 TI - Identification of two types of specific endothelin receptors in rat mesangial cell. AB - Two types of receptors specific for endothelin were identified using cross linking technique in cultured rat mesangial cells. The molecular weights of these receptors were approximately 58,000 and 34,000 by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The binding of radioiodinated-endothelin to its receptors was inhibited by excess of unlabeled endothelin, but not by nifedipine, nicardipine, verapamil, diltiazem, angiotensin II or [Arg8]-vasopressin. The endothelin binding proteins were solubilized with 1% digitonin and fractionated under non-denaturing conditions by gel filtration. Two endothelin binding peaks eluted at the positions corresponding to the molecular weights of 65,000 and 43,000. These observations indicate that there are two types of specific endothelin receptors in rat mesangial cells which are distinct from voltage dependent L-type calcium channel. PMID- 2548497 TI - Glycogen-bound type-1 phosphatase: isolation and dissociation of a complex containing undegraded G-subunit. AB - A high molecular mass type-1 phosphatase complex can be isolated from muscle glycogen particles by a fast procedure that preserves the glycogen-binding subunit of phosphatase called G from proteolysis. G can be dissociated from such complex by ion exchange chromatography on FPLC SI column, with recovery of unproteolyzed G completely separated from phosphatase catalytic subunit. PMID- 2548498 TI - The dephosphorylation of lens alpha-crystallin A chain. AB - The present communication reports the presence of a phosphoprotein phosphatase activity in bovine lens preparations which dephosphorylates alpha Ap, the phosphorylated form of alpha A, one of the alpha-crystallin polypeptides, in a Ca2+/calmodulin dependent manner. The activity was found in soluble preparations from epithelial cells but it could not be detected in similar preparations from fiber cells. A 60,000 Mr calmodulin binding polypeptide and a 15,000 Mr polypeptide found in the epithelial cell preparations comigrated in SDS-PAGE with the A and B subunits of bovine brain calcineurin (phosphoprotein phosphatase 2B) respectively. The 15,000 Mr was specifically recognized by an anti-bovine brain calcineurin antiserum. Bovine brain calcineurin was as effective in dephosphorylating alpha Ap as the lens preparations. Thus, it is likely that the activity present in the lens is related to this enzyme. The results indicate that the lens specific polypeptide alpha A may be subject to metabolic control through phosphorylation and dephosphorylation pathways regulated by cAMP and calcium respectively. Changes in the activities of these pathways appear to occur during differentiation of the lens epithelial cell and may be related to gene regulation during the differentiation process. PMID- 2548499 TI - Interaction of rat brain cytidylate cyclase with phospholipids. AB - The interaction of rat brain cytidylate cyclase with some phospholipids such as L alpha-phosphatidylcholine (PC), L-alpha-phosphatidylserine (PS), L-alpha phosphatidylethanolamine (PE) and L-alpha-phosphatidic acid (PA) was studied. Cytidylate cyclase activity of Triton X-100 - solubilized fraction was inhibited by PS, PE and PA, but not with PC. The addition of PC to the incubation mixture containing PS, PE or PA dose - dependently reversed the inhibition of enzyme activity by these phospholipids. Phospholipids showed similar effect on the intact membrane - bound enzyme. PC could reactivate the enzyme which was inactivated by deoxycholate treatment, suggesting that PC may be an important factor to reconstitute an active conformation of the enzyme. These findings indicate that cytidylate cyclase could be regulated by phospholipids constituting its microenvironment of the membrane. PMID- 2548500 TI - E-series prostaglandins activate cAMP-mediated potassium currents in follicle enclosed Xenopus oocyte. AB - Receptor-mediated responses to prostaglandins E1 and E2 are shown by electrophysiological methods in follicle-enclosed oocytes of Xenopus laevis. In voltage-clamped oocytes, prostaglandins E1 and E2 elicited an outward hyperpolarizing current. This outward membrane current was caused by an increase in K+ conductance. The prostaglandin-induced current was augmented by adenylate cyclase activator, forskolin, and by phosphodiesterase inhibitor, theophylline, indicating that adenosine 3', 5'-cyclic monophosphate (cAMP) is involved in activating the K+ current. The prostaglandin responses were either abolished or greatly reduced by removing follicular cells with collagenase, suggesting that prostaglandin receptors reside in the follicular cells. PMID- 2548501 TI - Parathyroid hormone (PTH) and PTH-like protein (PLP) stimulate interleukin-6 production by osteogenic cells: a possible role of interleukin-6 in osteoclastogenesis. AB - Osteogenic cells mediate PTH-stimulated osteoclastic bone resorption by a yet unidentified mechanism. We show that primairy rat osteoblast-like cells and the clonal osteogenic sarcoma cell line UMR-106 produce interleukin-6 (IL-6) and that bPTH(1-84) and synthetic hPLP(1-34) stimulate this production dose-dependently. With both peptides a close relation between IL-6 and cyclic-AMP production was found, though for PTH concentrations higher than 2.10(-8) M a clear dissociation was observed. Significant IL-6 activity was also detected in media of cultures of 17-day-old fetal mouse radii and metacarpals which was clearly stimulated by PTH. The source of IL-6 in these bone explants seems to be the osteogenic (cartilage) cells. Treatment of bone explants with IL-6 induced osteoclastic resorption which, however, depended on the bone resorption system used. This bone resorbing action of IL-6 is exerted probably through an effect on the formation of osteoclasts (osteoclastogenesis) rather than on the activation of already existing mature osteoclasts. We suggest that IL-6 produced by osteogenic cells may be a mediator in PTH-stimulated osteoclastic bone resorption. PMID- 2548502 TI - Functional endothelin/sarafotoxin receptors in the rat uterus. AB - Functional receptors for the peptides of the endothelin (ET) and sarafotoxin (SRTX) families were detected in the rat uterus. These receptors specifically bind 125I-SRTX-b (Bmax = 220 fmol/mg protein), as well as ET-1, ET-3 and SRTX-c (IC50's 10, 5, 300 and 780 nM, respectively). Activation of the uterine ET/SRTX receptors induced dose-dependent phosphoinositide (PI) hydrolysis and three typical contractile responses: 1) increase in the muscle tonic tension; 2) increase in frequency of the spontaneous rhythmic contractions; 3) decrease of relaxation in each spontaneous rhythmic cycle. All three effects appeared at doses as low as 0.5-1 nM. Dose responses yield ED50 values of 5.5, 30 and 680 nM for ET-1, SRTX-b and ET-3, respectively. SRTX-c was the least effective peptide in achieving decrease in relaxation. In view of these results, and since the uterine responses to the peptides were almost immediate and reversible, we suggest that the functional ET/SRTX receptor of the rat uterus that is coupled to PI hydrolysis may be of physiological significance. PMID- 2548503 TI - Time-course of Ca2+-induced insulin secretion from perifused, electrically permeabilised islets of Langerhans: effects of cAMP and a phorbol ester. AB - The pattern of insulin secretion from electrically permeabilised islets was studied in a perifusion system. Increases in intracellular Ca2+ stimulated insulin secretion in a dose-related manner, but the secretory response to Ca2+ was only transient, with permeabilised islets becoming rapidly insensitive to a stimulatory concentration of Ca2+. Cyclic AMP and the protein kinase C activator, phorbol 12-myristate 13-acetate (PMA), both stimulated Ca2+-induced insulin secretion from perifused permeabilised islets by increasing the maximum secretory response to Ca2+, and by maintaining elevated rates of secretion when the permeabilised islets had become insensitive to the stimulatory effects of Ca2+. These results suggest that cAMP and PMA may act partly by modifying the magnitude of the secretory response to Ca2+, and also by Ca2+-independent effects on the secretory mechanism. PMID- 2548504 TI - Intracellular regulatory system involving calpain and calpastatin. AB - Seven years have elapsed since the terms calpain and calpastatin were introduced. During these years, significant progress in research has been recorded. Thus, cloning and sequencing of cDNAs for calpains I and II and calpastatin have established amino acid sequences of these molecules. Structure-function relationship of calpastatin has been studied using mutated cDNAs expressed in E. coli. Interleukin 2 receptor-linked expression of calpastatin in HTLV-I-infected T-cells has been reported. Evidence for Ca2+-induced translocation of calpain to the cell membrane, followed by its autolytic activation, has been discussed. A great varieties of proteins such as several kinases, membrane and cytoskeletal proteins, and hormone receptors have been reported to be susceptible to calpains. This paper is to summarize our current knowledge on chemistry and biology of calpain and calpastatin and thereby to speculate the true function of calpains and their regulatory mechanisms. PMID- 2548505 TI - Inhibition of superoxide anion radical production by ebselen (PZ51) and its sulfur analogue (PZ25) in guinea pig alveolar macrophages. AB - The production of superoxide anion radicals by guinea pig alveolar macrophages is stimulated by the chemotactic peptide N-formylmethionyl-leucylphenylalanine and the protein kinase C activator phorbol-12-myristate-13-acetate. Both stimulations are completely and equipotently (IC50 = 20 microM) inhibited by the seleno organic compound ebselen. As the sulfur containing analogue, which is devoid of glutathione peroxidase-like activity, shows the same inhibitory activity towards superoxide anion radical production the observed effect of ebselen can not be explained by the described glutathione peroxidase-like activity. The observed inhibition is probably caused by an inhibition of protein kinase C or inhibition at a level distal to protein kinase C activation. PMID- 2548506 TI - Testosterone response in arginine vasopressin desensitized immature rat testis. AB - Direct injection of arginine vasopressin into immature rat testis inhibited basal testosterone synthesis. Simultaneous injection of arginine vasopressin with luteinizing hormone, norepinephrine or cholera toxin inhibited these agonists - induced testosterone response. In arginine vasopressin - desensitized testis, cAMP response to luteinizing hormone, norepinephrine and cholera toxin was not disturbed. However, testosterone response to luteinizing hormone, norepinephrine or cholera toxin was drastically reduced in arginine vasopressin-desensitized testis. This shows that the increased cAMP generated by luteinizing hormone, norepinephrine or cholera toxin in arginine vasopressin desensitized testis did not cause increase in steroidogenesis. This could be due to a lesion in steroidogenic pathway beyond cAMP generation caused by arginine vasopressin. PMID- 2548507 TI - Blockage of morphological transformation of chick embryo fibroblasts infected with Rous sarcoma virus by N-myristoyl glycinal diethylacetal in vitro. AB - N-Myristoyl glycinal diethylacetal strongly inhibited morphological transformation of chick embryo fibroblasts infected with a temperature-sensitive mutant (tsNY68) of Rous sarcoma virus. Myristoylated or nonmyristoylated pp60v src, which were expressed in tsNY68-infected cells in the absence or presence of the compound, were identified separately by fluorography or immunoblotting analysis after sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the [3H]myristate-labeled cell lysate. The results taken together suggest that the blockage of morphological transformation was caused by prevention of protein myristoylation of the transforming protein pp60v-src. PMID- 2548508 TI - Purification and some properties of a 2Fe ferredoxin in Pseudomonas ovalis. AB - A [2Fe-2S] ferredoxin was found in Pseudomonas ovalis which was grown in a medium supplemented with glucose and ammonium sulfate. The molecular weight of the 2Fe ferredoxin was estimated to be 13,000. It contained 2.2 gramatoms of non-heme iron and 2.3 gramatoms of acid-labile sulfur per mole protein. The absorption and circular dichroism spectra were characteristic of those of [2Fe-2S] type ferredoxins, especially adrenodoxin and putidaredoxin. The electron paramagnetic resonance spectrum of the reduced protein showed an axial symmetry (g = 2.020, g = 1.939). The amino acid composition was determined. PMID- 2548509 TI - Distribution of glycosyltransferase activities in different compartments of mitochondria. AB - The liver mitochondria were submitted to a first swelling which allowed to get outer membranes. The mitoplasts obtained in these conditions were subject to a second swelling. The separation of submitochondrial membranes on a discontinuous sucrose gradient revealed three membrane fractions, an outer membrane rich fraction, an inner membrane rich fraction and a fraction enriched with contact sites between the two membranes. The various glycosyltransferase systems involved in the biosynthesis of N-glycoproteins were located in these fractions. PMID- 2548510 TI - Identification and characteristics of a novel mitochondrial 5'-nucleotidase in rat liver. AB - In rat liver mitochondria there exists an AMP-dephosphorylating activity which converts external 5'-AMP to adenosine. It exhibits a pH optimum of 7.5 and a Km(AMP) of 0.085 mM. Furthermore, this activity is stimulated by magnesium (Km = 0.5 mM) and seems to be not affected by low concentrations of ATP or ADP. From the characteristics of the enzyme the existence of a 5'-nucleotidase in rat liver mitochondria which is localized on the outer surface of the inner mitochondrial membrane was concluded. The enzyme may be important for the production of cellular adenosine. PMID- 2548511 TI - Salt stimulation of the activation of latent protein phosphatase, Fc.M, by Mn++ and by Mn/trypsin. AB - The activation of porcine heart latent protein phosphatase (Fc.M) by pretreatment with Mn++ followed by trypsin (Mn/trypsin) can be stimulated 2.5-fold by including NaCl or KCl in the activation mixtures. The salts also stimulated the activation of the enzyme by Mn++ to the same level as that obtained by Mn/trypsin pretreatment in the absence of salt. The presence of salt in both the Mn++ and Mn/trypsin activations decreased the Mn++ requirement 10-fold in each case. Treatment of latent Fc.M by Mn/trypsin in the presence of 0.2 M NaCl or KCl offers a convenient method of expressing the full potential activity of the protein phosphatase. PMID- 2548513 TI - Regulation of purine deoxynucleoside phosphorylation by deoxycytidine kinase from human leukemic blast cells. AB - The kinetics and regulation of nucleoside phosphorylation by highly purified human deoxycytidine kinase from leukemic lymphoblasts were studied. The phosphorylation of purine nucleosides by this enzyme showed sensitivity to the endogenous inhibitors dCTP and UDP three times greater than the phosphorylation of dCyd. Examination of nucleotide pools in human T and B lymphoblasts disclosed that the levels of dCTP and UDP in these cells were sufficient to regulate kinase activity. The enhanced sensitivity of the kinase to dCTP and UDP was related to its reduced ability to interact with purine nucleosides. Comparison of the phosphorylation kinetics for pyrimidine and purine dideoxynucleosides used in antiviral therapy showed that the purine nucleosides were at least 50-fold less efficient as enzyme substrates. These results suggest that the phosphorylation of pharmacologically active purine nucleosides by deoxycytidine kinase is regulated by cellular nucleotide pools. PMID- 2548512 TI - 36Cl- flux measurements on GABAA receptor-activated chloride exchange. Multiple mechanisms of the chloride channel inactivation. AB - The technique of radiotracer 36Cl- influx in primary culture of rat cerebellar granule cells was applied to study the mechanism of inactivation of the GABAA receptor-activated chloride channel. During sustained application of GABA, muscimol and THIP the specific bicuculline-sensitive 36Cl- influx tends to decline with time. The sequence in decay half-time is GABA less than muscimol less than THIP. Diazepam accelerates the rate of decay of the peak response to GABA. (-)-Baclofen enhances the rate of decline of the response to muscimol in a dose-dependent manner. Treatment of the cells with pertussis toxin antagonized the effect of (-)-baclofen. It is concluded that rat neonatal cerebellar neurons maintained in tissue culture exhibit complex inactivation of the GABAA channel, indicating some interaction with the GABAB receptor system. PMID- 2548514 TI - [Plasmids pMB123 and pMB124--vectors for obtaining subfragments of DNA with random "sticky" ends]. AB - For preparing a DNA fragment with unique protruding ends, plasmid vectors pMB123 and pMB124 were constructed by inserting a synthetic polylinker into plasmid pUR222 at the EcoRI-PstI sites. The polylinker contains two FokI and HgaI sites at its ends in opposite orientation flanking a combination of SalGI, AccI, HindII, HindIII (the latter site is absent from pMB124) and BamHI sites. DNA fragment cloned at the SalGI and BamHI sites can be regenerated by either FokI or HgaI treatment, the SalGI and BamHI sites being deleted from the cloned sequence. Fragments coding for parts of human interleukin-2 were cloned in these vectors. PMID- 2548515 TI - [Human B lymphotropic virus (HBLV) or human herpesvirus (HHV-6)]. AB - HHV.6 (or HBLV) herpesvirus was isolated in 1986. This virus has a specific tropism for T cells. It is ubiquitous. Most subjects are infected in early childhood. HHV.6 is responsible for exanthema subitum. Its possible involvement in the aggravation of immunodeficiency syndromes and its action on malignant haematologic disorders, has been suggested but has not been clearly demonstrated. PMID- 2548516 TI - Effect of Ca2+-homopantothenate and mild hypoxia on some enzyme activities evaluated in subcellular fractions from different rat brain regions. AB - The effect of Ca2+-homopantothenate (HOPA) treatment (250 mg/kg for 5 d) has been studied by evaluating the specific activity of enzymes related to: glycolytic pathway (hexokinase, phosphofructokinase, pyruvate kinase, lactate dehydrogenase), tricarboxylic acid cycle (citrate synthase, malate dehydrogenase), mitochondrial electron transfer chain (succinate dehydrogenase, cytochrome oxidase), NADH redox state (NADH cytochrome c reductase), acetylcholine metabolism (acetylcholinesterase), and glutamate metabolism (glutamate dehydrogenase). The enzymatic activity assays were performed on homogenate in toto, nonsynaptic mitochondria and synaptosomes isolated from: cerebral cortex, hippocampus, striatum, hypothalamus, medulla oblongata, and cerebellum of normoxic rats and rats submitted to intermittent normobaric hypoxia (90:10, N2:O2). In normoxic rats, HOPA was unable to induce any modification. Hypoxia per se induced a decrease in the activity of synaptosomal cytochrome oxidase in cerebral cortex, hippocampus, and cerebellum. PMID- 2548517 TI - [Parvovirus infections in humans]. AB - The Human Parvovirus (HPV) like other Parvovirus is a single strand DNA virus with autonomous replication which measures 23mm. Infection with this virus is followed by a non-specific viral syndrome during the prodrome, leading to viremia, which may be followed by arthropathy and/or different kind of rash including the syndrome called erythema infectiosum. It has also been related to an increase in the number of spontaneous abortion in pregnant women with acute infection; and it is the etiology of the aplastic crisis in patients with hemolytic anemias. Many other Parvovirus serologically different from HPV are present in stools and are responsible for acute infectious non bacterial gastroenteritis in people more than 5 years old. PMID- 2548518 TI - Flowing by the waste-side: the emerging national policy on medical waste. PMID- 2548519 TI - Morphologic and immunocytochemical studies of bronchioloalveolar carcinoma at Duke University Medical Center, 1968-1986. AB - Between 1968 and 1986, the tumor registries at Duke University Medical Center and Durham VA Medical Center accumulated a total of 193 patients with a diagnosis of bronchioloalveolar carcinoma (BAC). All available histologic sections of primary lung tumors and all available respiratory and pleural cytologic material were reviewed for 135 cases having an initial histologic diagnosis of BAC and no history of a primary nonpulmonary adenocarcinoma. Thirty-nine cases showed a pure BAC pattern in histologic sections; 76 showed a dominant BAC pattern with focal areas of fibrosis and acinar differentiation; 16 were carcinomas with a focal BAC pattern; and 4 were adenocarcinomas lacking a BAC pattern. Of the 115 cases with at least a dominant BAC pattern, 51 showed predominantly mucinous differentiation while 64 showed predominantly nonmucinous differentiation. Adequate cytologic material was available for review from 111 patients. For cases having at least a dominant BAC pattern, tumor cells were present in 77 of 172 adequate sputums (44.8%), 36 of 133 bronchoscopy specimens (27.1%), 15 of 18 needle aspirates (83.3%) and 14 of 15 pleural fluids (93.3%). Of all patients in this group, 60.2% had at least one specimen positive for malignancy. No cytologic features clearly distinguished adenocarcinomas having only focal bronchioloalveolar differentiation from those with a pure or dominant BAC pattern. A significant degree of overlap was observed in the cytologic features of mucinous and nonmucinous tumors. Histologic sections from 19 mucinous and 16 nonmucinous tumors were stained with monoclonal antibody B72.3: all showed some staining, with no significant difference in degree of staining between the two groups. This suggests that expression of the tumor-associated glycoprotein TAG-72 is independent of mucinous differentiation. PMID- 2548520 TI - Surgical adjuvant therapy of non-small-cell lung cancer. AB - Results of several studies by the Lung Cancer Study Group have shown that postoperative adjuvant chemotherapy enhances survival following surgery for lung cancer. The 18-month disease-free survival almost doubled in one study using cyclophosphamide, doxorubicin, cisplatin (CAP) chemotherapy postoperatively. The recurrence rate remained significant, however. Patients with more advanced resectable disease seem to benefit most from postoperative chemotherapy. Results also suggest that CAP delays recurrences more effectively in patients with nonsquamous vs. squamous lung carcinoma. There has been considerable interest in the use of preoperative adjuvant therapy as well. Findings from studies of preoperative or induction therapy--either chemotherapy alone or in combination with radiation therapy--have shown high response rates and that patients with unresectable disease can be converted to technically resectable. Although preoperative therapy can cause difficulties with surgical dissection, surgical morbidity is acceptable. Preoperative chemotherapy and radiotherapy followed by surgical resection clearly eliminates local recurrence. Systemic recurrences remain a significant problem. The evidence, as yet, does not indicate that preoperative adjuvant therapy prolongs survival. PMID- 2548521 TI - Multiple forms of parathyroid hormone-like proteins in a human tumour. AB - The extensive chromatographic characterization of four parathyroid hormone (PTH) like proteins in a human bronchial carcinoid tumour associated with humoral hypercalcaemia and severe osteitis fibrosa is described. PTH-like bioactivity was detected in acetic acid extracts of the tumour using an in-vitro osteo-sarcoma cell bioassay. The active tumour proteins were positively charged at physiological pH and had apparent Mr of approximately 29,000, 16,000, 4000-9000 and less than 4000. The proteins were immunologically distinct from PTH, but each stimulated PTH-sensitive adenylate cyclase in cultured osteoblastic cells. There was no evidence of PTH gene expression by the tumour. These proteins represent different molecular forms of PTH-related protein. PMID- 2548523 TI - Bradykinin stimulates phosphoinositide metabolism and prolactin secretion in rat anterior pituitary cells. AB - Bradykinin stimulated prolactin secretion from monolayer cultures of rat anterior pituitary cells, the stimulation being greater from the cells of male rats. This stimulated secretion was accompanied by a rise in total inositol phosphate accumulation, suggesting that the action of bradykinin is mediated by phosphoinositide hydrolysis. The increase in inositol phosphate accumulation was biphasic; a further sharp rise occurred when the concentration of bradykinin exceeded 1 mumol/l. This may indicate that bradykinin acts on other cell types in the pituitary gland. Bradykinin had no effect on growth hormone secretion from cells of normal pituitary glands, or on prolactin secretion and phosphoinositide metabolism in GH3 rat pituitary tumour cells. Bradykinin receptor antagonists (both B1 and B2) had no effect on either bradykinin-stimulated inositol phosphate accumulation or prolactin secretion. Kallikreins, the enzymes responsible for the generation of kinins, are known to be present in the adenohypophysis. Therefore, the results presented here would suggest that kinins may have a role as paracrine agents in the pituitary gland. PMID- 2548522 TI - Immunological distribution of one form of insulin-like growth factor (IGF) binding protein and IGF peptides in human fetal tissues. AB - Insulin-like growth factors (IGFs) are expressed by, and are biologically active on, human fetal cells. The mitogenic actions of IGF-I are modulated by the 21-41 kDa class of IGF-binding proteins (IGF-BPs). Using a rabbit anti-human IGF-BP antibody raised against a highly pure 26 kDa IGF-BP derived from amniotic fluid, we have compared the cellular location of IGF-BP and IGF peptides in tissue sections from prostaglandin-induced human abortuses of 14-16 weeks of gestation. The monoclonal and polyclonal antibodies used were raised against human IGF-I, but did not distinguish between IGF-I and IGF-II. Positive staining for IGF-BP was seen in every tissue except brain, spleen and thyroid. With the exception of skin, the cellular distribution of IGF-BP was similar to that of IGF peptides. Strong immunostaining was found in hepatocytes, hepatic erythropoietic cells, pulmonary epithelium, the tubular epithelium of kidney, intestinal epithelia, the fetal adrenal cortex and cardiac and skeletal muscle fibres. In skin, IGF-BP was located throughout the dermis and in the germinal layer of the epidermis. IGF peptide in skin was restricted to the deeper dermal layers. In the tibial epiphyseal growth plate both IGF-BP and IGF peptide were located in chondrocytes throughout the proliferation and hypertrophic zones. The similarity in distribution of IGF-BP and IGF peptides in fetal tissues suggests that the latter may exist predominantly complexed to IGF-BP in or on the surfaces of cells in vivo. The distribution of IGF-BP may define the sites of biological action of IGF peptides. PMID- 2548524 TI - Methionine-oxidized horse heart cytochrome c. III. Ascorbate reduction and the methionine-80-sulfur-iron linkage. AB - The ascorbate reduction of the CT-cytochromes--two chemically generated forms of horse heart cytochrome c, FIII and FII, with both methionines, 80 and 65, as methionine sulfoxides, no iron-sulfur linkage, and potentiometric and physiological oxidoreduction properties distinct from those of the native protein and one another (J. Pande et al., 1987)--has been investigated using a stopped flow technique. The reaction was monitored at 550 nm, and studies were conducted in 10 mM phosphate + 0.17 M NaCl buffer, pH 7.4. Both CT-cytochromes are reduced by triphasic profiles, a faster and an intermediate ascorbate-dependent reaction and a slow, ascorbate-independent process. Both CT-cytochromes contain three molecular forms in slow equilibrium, two reducing directly by reaction with ascorbate and a third through conversion to one of the reducible forms. Like the reaction of the native protein, the ascorbate dependence of both the rapid and the intermediate process is nonlinear, approaching saturation values at high concentrations. The ascorbate profiles of the pseudo-first-order reduction constants are typical of the model for the reduction reaction of the unmodified protein, binding followed by a first-order reduction reaction (Myer et al., 1980; Myer and Kumar, 1984), but with distinct kinetic parameters, the first-order reduction constants and the protein-ascorbate stability constants. It has been concluded that the functional-conformational differences between the two CT cytochromes are not operational to any significant extent in the reduction reaction with ascorbate. The methionine-80-sulfur-iron linkage of the protein is not a crucial requirement for the ascorbate reduction of the protein. The mechanism of the reaction in the main is also insensitive to the replacement of Met-80-S from heme coordination and/or the associated conformational oxidoreduction properties of the protein. Of the two aspects of the reaction, the efficiency of the electron-transfer reaction and the stability of the ascorbate dianion-protein complex, the former is dependent on the integrity of the structural-conformational state of the molecule. PMID- 2548526 TI - Mutational specificities of environmental carcinogens in the lacl gene of Escherichia coli. II: A host-mediated approach to N-nitroso-N,N-dimethylamine and endogenous mutagenesis in vivo. AB - An intrasanguineous host-mediated assay was used to determine the mutational specificity of the hepatocarinogen N-nitroso-N,N-dimethylamine metabolized in vivo. A total of 114 forward mutations in the lacl gene of Escherichia coli reisolated from the livers of treated Swiss albino mice were characterized at the DNA sequence level. Consistent with the methylating ability of this compound and the demonstrated mutagenic specificity of O6-methylguanine, the predominant mutation was the G:C----A:T transition. These were recovered, on average, seven times more frequently at guanines flanked (5') by a purine residue than at those preceded by a pyrimidine residue--a specificity similar to that reported for many direct-acting SN1 alkylating agents. This nitrosamine appears to be distinguished from related N-nitroso methylating compounds by the induction of additional mutational events. Here, the exceptions consisted of four A:T----G:C transitions, four A:T site transversions, and a single G:C----T:A transversion. In addition, the DNA sequence alterations of 34 I- mutants of E. coli reisolated from otherwise untreated mice were identified. The predominant mutation was the G:C--- A:T transition, which accounted for almost half of all background mutations. The sites at which these mutations were recovered appear to indicate that some of these mutations may have arisen as a result of an accelerated rate of cytosine deamination. These data suggest that many of the additional "spontaneous" mutations observed under in vivo conditions resulted from genotoxic events occurring during the host-defense (immune) reaction. PMID- 2548527 TI - Malignant transformation of human bladder epithelial cells by DNA transfection with the v-raf oncogene. AB - Transfection of the v-raf oncogene into immortalized, nontumorigenic human bladder epithelial cells resulted in the isolation of two tumorigenic transformants. Both were identified as human and of the same origin as the parent cell line by human leukocyte antigen typing and Southern blot analysis. Both the primary tumorigenic transfectants and the cell lines established from the induced tumors expressed v-raf mRNA and v-raf protein. In both tumorigenic transformants the level of c-myc mRNA was enhanced compared with that of the parent cell line. PMID- 2548525 TI - Comparative X-ray crystallographic evidence for a beta-bend conformation as the active structure for peptide T in T4 receptor recognition. AB - A sequence similarity has been found between two segments of endothiapepsin (acid proteinase, 2APE), bovine pancreatic ribonuclease A, and peptide T, a segment of the gp120 protein of human immune deficiency virus (HIV), which has been implicated in blocking viral attachment to the T4 receptor. The two similar sequences of the acid proteinase enzyme are Leu-Ile-Asp-Ser-Ser-Ala-Tyr-Thr (residues 169-176) and Tyr-Thr-Gly-Ser-Leu-Asn-Tyr-Thr (residues 175-182). Since the X-ray crystallographic structures of the acid proteinase and ribonuclease are known, it has been possible to determine whether the three-dimensional structures of the segments are similar. Portions of both the segments of acid proteinase are directly superimposable on the structure of the RNase A 19-26 segment. The fact that the three similar sequences from two completely unrelated proteins give rise to almost identical structures raises the possibility that these segments may be involved in nucleating the folding of these proteins. In addition, this provides further support for the concept that the octapeptide sequence of peptide T of HIV, which is also similar in sequence to the 19-26 sequence of RNase A, is also structurally similar to these residues, which adopt a beta-bend conformation. Furthermore, comparison of similarities and differences in the structure of these similar sequences provides an explanation for alterations in the biological activity of various truncated or substituted derivatives of peptide T and additional confirmation of the structural requirements for peptide T in T4 receptor recognition. PMID- 2548528 TI - Chromatin structure and transcriptional regulation of human papillomavirus type 18 DNA in HeLa cells. AB - Mapping analysis of the nucleosomal organization of integrated human papillomavirus type 18 (HPV18) DNA in HeLa cells reveals a very prominent nuclease-hypersensitive site within the viral noncoding regulatory region that harbors transcriptional control sequences and coincides with most of the 5' ends of the cytoplasmic early mRNAs. Moreover, it is shown that the conserved coamplified 5' cellular flank, common to all HPV18 copies in HeLa cells and located close to the virus-cell integration site, also contains several distinct hypersensitive sites, accessible not only to DNase I but also to restriction enzymes. Nuclear run-on analysis in isolated HeLa nuclei demonstrates the occurrence of nascent transcripts covering the cellular flank (the late and the viral noncoding regulatory region), indicating that a cellular promoter, marked by the hypersensitive sites, cooperates with the viral control region in generating the HPV18 transcripts. Cycloheximide treatment of HeLa cells results in a reduction of the cytoplasmic steady-state level of the 3.5-kb mRNA corresponding to the viral E6, E7, and parts of the E1 open reading frames (ORFs), whereas the expression of the 1.6-kb transcript corresponding only to the E6 and E7 ORFs is not influenced. Nuclear run-on analysis carried out after the cycloheximide chase reveals that the distribution of nascent transcripts spanning the viral E6, E7, and parts of the E1 region is substantially decreased. In contrast to this finding, an even, pronounced increase of the elongation rate of those transcripts, which cover the cellular flank, the late and the viral noncoding regulatory region was noted indicating a different involvement of regulatory factors in the activity of both promoters. PMID- 2548529 TI - Induction of metallothionein mRNA by tumor promoters in mouse skin and its constitutive expression in papillomas. AB - A single topical application of 12-O-tetradecanoylphorbol-13-acetate (TPA) was found to induce mRNA of a metallothionein (MT) gene or genes in the skin of Sencar mice, and papillomas produced by repeated applications of TPA were shown to have elevated levels of MT mRNA. Induction of MT mRNA was maximal 4-8 h after application of TPA and returned to the control level 24 h later. A dose-dependent increase of MT mRNA was observed with doses of TPA of 1-5 micrograms. Of the other promoters tested, phorbol-12, 13-didecanoate, mezerein, and the ionophore A23187 also induced MT mRNA, but 4-O-methyl-TPA and benzoyl peroxide did not. Phorbol and 4 alpha-phorbol-12,13-didecanoate, which are not promoters, also did not induce MT mRNA. Retinoic acid and 1 alpha, 25-dihydroxyvitamin D3, inhibitors of tumor promotion, did not induce MT mRNA themselves or inhibit the induction of MT mRNA by TPA. In C57BL/6 promotion-resistant mice, TPA caused only slight induction of MT mRNA. These data suggest a correlation between induction of MT mRNA and epidermal hyperplasia. The constitutive elevation of MT mRNA levels in papillomas may be due to the loss, during the process of tumor promotion, of some mechanism regulating MT gene expression. PMID- 2548530 TI - [Cell proliferation of the intestinal mucosa in patients with familial adenomatosis coli. Autoradiography study]. AB - In connection with routine endoscopic examinations of 31 patients with familial adenomatosis coli and 28 healthy kindreds of patients with familial adenomatosis coli we investigated by autoradiography the in vitro incorporation of 3H thymidine into bioptic mucosal particles from colon and adenomas. Within the 5 year follow-up period there was found a significantly increased labelling index of polyps and the surrounding mucosa in patients with polyposis independing on the time elapsed after operation. In 12 of 28 kindreds the histological normal colonic mucosa showed an increased labelling index too. The prognostic value of these results must be proved by further controls. PMID- 2548532 TI - Decreased reactivity to the anticonflict effect of diazepam in perinatally undernourished rats. AB - Offspring of rats were submitted to a protein deprivation dietary treatment from the third week of gestation until 50 days of age, and later nutritionally rehabilitated for at least 90 days. In the punished lever-pressing conflict test, undernourished animals exhibited a decreased reactivity to the anticonflict effect of a 3 mg/kg dose of diazepam as compared to control animals. This decreased reactivity to diazepam in a shock-induced conflict test indicates that functional alterations in the GABAergic transmission might contribute to the state of hypersensitivity to stressful or aversive situations present in undernourished animals. PMID- 2548531 TI - Effects of opiate antagonists and their quaternary analogues on nucleus accumbens self-stimulation. AB - Naloxone and naltrexone were compared with their quaternary analogues naloxone methobromide and naltrexone methobromide for efficacy in suppressing intracranial self-stimulation behavior. These quaternary analogues effectively block opiate receptors in the periphery, but since they do not readily cross the blood-brain barrier they have little effect on central receptors. Rats with electrodes in the nucleus accumbens were trained to self-stimulate in daily 60-min sessions. Naloxone (0.2, 2.0 and 20 mg/kg) and naltrexone (20 mg/kg) potently suppressed self-stimulation behavior. In contrast, neither naloxone methobromide (0.2 and 20 mg/kg) nor naltrexone methobromide (20mg/kg) had any significant effects on this behavior. These results suggest that blockade of peripheral opiate receptors alone is insufficient to suppress self-stimulation, and therefore support the idea that opiate antagonists suppress self-stimulation by blockade of central receptors that mediate reinforcement. PMID- 2548533 TI - Procedures that produce context-specific tolerance to morphine in rats also produce context-specific withdrawal. AB - Rats previously injected with morphine in the presence of a distinct environment (paired animals) were more tolerant to the analgesic effects of morphine in that environment than were rats previously injected with morphine in another environment (unpaired animals). When injected with saline instead of morphine in the distinct environment, paired animals were more reactive to pain (hyperalgesic) than unpaired animals, but no more reactive to pain than animals never given morphine. More important, the paired animals also exhibited more withdrawal symptoms (wet dog shakes, genital licking, circling, rearing, and defecation) during abstinence and naltrexone-precipitated withdrawal in the distinct environment than did the unpaired and saline animals. Thus, procedures that are capable of producing context-specific opiate tolerance are also capable of producing context-specific opiate withdrawal. PMID- 2548534 TI - Central mediation of naloxone-induced anorexia in the ventral tegmental area. AB - Central naloxone injections were used to show that endogenous opioids in the ventral tegmental area (VTA) regulate consumption of palatable foods. Peripheral injections of naloxone were more effective in reducing the consumption of a sweet solution in normally fed rats than in animals maintained at 85% of their free feeding body weight. A dose of 10 micrograms/side naloxone injected into the VTA reduced consumption in normally fed rats, whereas a dose of 25 micrograms/side did the same in food-restricted animals. The inactive isomer, (+) naloxone, did not reduce consumption; the effect has chemical specificity. Naloxone injected 0.5, 1.0, or 2.0 mm above the VTA did not reduce consumption; the effect has anatomical specificity. Naloxone effectively decreases the eating of palatable foods, but not eating for survival. This has important implications for the use of opioid antagonists in weight-loss programs. PMID- 2548535 TI - Antibody-dependent cellular cytotoxicity is directed against both the gp120 and gp41 envelope proteins of HIV. AB - To define the target antigens for antibody-dependent cellular cytotoxicity (ADCC), assays were performed using affinity-purified human immunoglobulin (Ig) or polyclonal rabbit sera directed against specific proteins of HIV. ADCC was not found using affinity-purified anti-core (p25) human Ig or sera obtained from rabbits hyper-immunized with recombinant p25. However, when affinity-purified human Ig or rabbit antisera specific for the envelope glycoproteins, gp120 or gp41, were used in ADCC assays, killing of HIV-infected cells was observed. These results indicate that antibodies in the infected individual that mediate ADCC are directed against both the gp120 and gp41 HIV envelope proteins and not against the viral core protein. PMID- 2548536 TI - Urinary neopterin concentrations in rhesus monkeys after infection with simian immunodeficiency virus (SIVmac 251). AB - Two rhesus monkeys were infected with SIVmac 251. Elevated urinary neopterin concentrations were observed as the first sign of infection. Virus-specific antibodies were detected 14 days after infection, when neopterin concentrations were already decreasing. The neopterin levels of one animal remained elevated and the virus was repeatedly isolated. Urinary or serum neopterin concentrations appear to be early markers for SIV infection and viremia in rhesus monkeys. PMID- 2548537 TI - Statistics from the World Health Organization and the Centers for Disease Control. PMID- 2548538 TI - Alterations of the eustachian tube, middle ear, and nose in rhinovirus infection. AB - This study measured the changes in 64 ears of 32 adult volunteers in eustachian tube function by the nine-step test, middle ear pressure by tympanometry, and nasal patency by active posterior rhinomanometry for 18 days following type 39 or Hanks rhinovirus infection. Abnormal measures were limited to the 75% of subjects (24) with clinical illness (colds) defined by symptom scores. Two days after infection, tubal function was present in only 50% of ears (48) and 20% of persons (5), middle ear underpressures of less than -50 mm H2O were measured in 50% of volunteers (12), and decreased nasal patency was observed in 54% of those with colds (13). These changes resolved 6 to 10 days after challenge. These results support a causal relationship between viral upper respiratory tract infection eustachian tube obstruction and abnormal middle ear pressure. PMID- 2548539 TI - Surgical management of nasopharyngeal angiofibroma involving the cavernous sinus. AB - Involvement of the cavernous sinus by juvenile nasopharyngeal angiofibroma represents a therapeutic challenge. We present our experience over the past 5 years with the surgical management of six cases of juvenile nasopharyngeal angiofibroma involving this site. Three of six patients had involvement of the medial aspect of the cavernous sinus and tumor was removed using a midline extracranial approach. Of three remaining patients, two had invasion of the medial and inferior margin of the cavernous sinus and one represented a recurrent lesion. The tumor in these three cases was resected using a combined frontotemporal and lateral infratemporal fossa approach. An extracranial recurrence occurred in one patient, and the remaining five patients have had no evidence of recurrent disease 12 to 71 months following surgery. Morbidity has been limited to trismus, facial hypesthesia, and serous otitis media. PMID- 2548540 TI - Polypeptide composition of paired helical filaments. AB - Immunocytochemical studies using antibodies to cytoskeletal proteins have provided conflicting data on the components of paired helical filaments (PHF), due solely to immunological cross-reactivities. To avoid such ambiguity, we developed a protein chemical approach to the identification of the PHF components. After treatment with formic acid, PHF were digested with lysylendopeptidase and the resultant peptides were separated by HPLC. All major peaks were analysed for their amino acid compositions and sequences. From the PHF digest, proteolytic fragments of ubiquitin, tau and beta protein were sequenced. Ubiquitin in PHF appears to be in a conjugated form, while its target protein remains unidentified. Tau is integrated into PHF at the site of its carboxyl third. The presence of beta protein fragments is best interpreted as being due to contamination of amyloid filaments in the PHF preparation. Thus, ubiquitin and tau are the two definite components of PHF. PMID- 2548542 TI - Histological evaluation of prostatic cancer. 1. Reproducibility of tumour type. AB - It has been claimed that the histological type of prostatic carcinoma bears influence on the prognosis of the patient, and that different treatments have to be considered for different types of tumours. The primary aim of this study was to evaluate the reproducibility of the histological criteria for the ordinary acinic adenocarcinoma and ductal adenocarcinoma. From 85 prospective and consecutive cases of prostatic carcinoma all histological slides were examined on two independent occasions by each of 4 pathologists. The overall inter observer agreement and mean intra observer agreement were 0.69 and 0.74, respectively. The kappa coefficients of these inter and mean intra observer agreements were 0.39 (S.E. = 0.02) and 0.48 (S.E. = 0.08), respectively. Considering a dichotomous classification of pure acinic versus all other types of prostatic carcinoma, an overall inter observer agreement of 0.78 and mean intra observer agreement of 0.84 were found. The kappa coefficients of this dichotomous classification were 0.51 (S.E. = 0.03, inter) and 0.61 (S.E. = 0.11, intra), respectively. It is concluded that the criteria for differentiation between different histological types of prostatic carcinoma need to be improved. PMID- 2548541 TI - Superoxide production of peritoneal macrophages in experimental gram-negative sepsis; influence of in vitro and in vivo supplements of zinc. AB - Although zinc is essential for the optimum function of the immune system, there is some controversy regarding treatment with zinc during acute infections where low serum zinc levels are often recorded. The aim of the present study was to investigate the influence of in vitro and in vivo zinc supplementation on the potentially toxic metabolic activity of peritoneal macrophages during infection. Rats were made septic by implanting a gelatin capsule containing known amounts of E. coli, and Bacteroides fragilis into the abdomen. Peritoneal macrophages were harvested by peritoneal lavage 72 hours after the induction of sepsis. Superoxide release was measured after stimulation with phorbol myristate acetate (PMA) or serum treated zymosan (STZ). Macrophages from septic rats released significantly higher amounts of superoxide compared with macrophages from sham operated controls after stimulation with both PMA and STZ. Following in vitro supplementation, zinc inhibited the superoxide production of macrophages harvested from septic rats after stimulation with both PMA and STZ. In vivo supplementation with zinc resulted in increased superoxide production from septic macrophages when stimulated with STZ, whereas stimulation with PMA produced no significant changes. Thus, in vitro incubation inhibited the superoxide production of peritoneal macrophages in intraabdominal sepsis, whilst in vivo administration of zinc produced no such effect, and the effect seemed to vary depending on the stimuli used to initiate the respiratory burst. PMID- 2548544 TI - Intracellular calcium and cell function. PMID- 2548543 TI - Mediastinal malignant carcinoid with Cushing's syndrome: immunohistochemical and ultrastructural study. AB - A case of thymic atypical carcinoid with Cushing's syndrome and unfavorable clinical course is reported. Immunohistochemical analysis reveals distinct staining of tumor cells for ACTH, neuron-specific enolase, chromogranins (CG) and S-100 protein and with PHE-5 monoclonal antibody. At an ultrastructural level, the cells are undifferentiated with only a few neurosecretory granules. In the present case, immunohistochemical stainings for CG and with PHE-5 antibody seem reliable diagnostic tools, easily demonstrating the neuroendocrine nature of the neoplasm. NSE immunoreactivity can be an additional criterion. S-100-positive cells, which are present throughout the tumor, recall 'sustentacular cells', described in other neuroendocrine tumors. PMID- 2548545 TI - The human immunodeficiency virus long terminal repeat is preferentially expressed in Langerhans cells in transgenic mice. AB - Four lines of transgenic mice containing the HIV LTR linked to the bacterial gene encoding chloramphenicol acetyltransferase (CAT) were constructed. In each line, a characteristic tissue pattern of CAT expression was observed with detectable levels present in the eye, heart, spleen, thymus, and tail. Low levels of CAT were present in circulating lymphocytes, but CAT activity in these cells could be augmented following treatment with the mitogen phytohemagglutinin (PHA). Likewise, CAT expression was present at only low levels in circulating monocytes, but higher levels of CAT were observed in macrophages grown in the presence of various cytokines (CSF-1, GM-CSF, IL-1 alpha, IL-4, and IL-2). Furthermore, Langerhans cells recovered from skin showed higher levels of CAT activity than those observed in other cells of monocyte-macrophage lineage. These results indicate that LTR-CAT expression in cells of monocyte-macrophage lineage may increase in proportion to the degree of differentiation of these cells. These animals may be useful in the study of cell-specific determinants of LTR-directed gene activity and may serve to identify exogenous cofactors that promote the progression of HIV-related disease in vivo. PMID- 2548546 TI - Accessibility of the highly conserved amino- and carboxy-terminal regions from HIV-1 external envelope glycoproteins. AB - Amino- and carboxy-terminal extremities of the envelope external glycoproteins are regions that have remained highly conserved between human immunodeficiency viruses HIV-1 and HIV-2. The corresponding peptides have been synthesized and their structure and function analyzed. Circular dichroism spectra showed evidence of alpha helical conformation when the peptides were dissolved in the nonpolar solvent trifuoroethanol. These two regions are indeed exposed on the molecule because they were accessible to their respective specific antibodies on the native gp160 precursor or processed gp120 glycoproteins of HIV-1. Neither the peptides nor rabbit or human antibodies directed against the N- and C-terminal peptides interfered with the interaction between HIV-1 external glycoprotein gp120 and its CD4 cellular receptor. Taken together, these results indicate that N- and C-terminal regions of gp120 are accessible on the quaternary structure of the virion as well as on the soluble form of gp120 and that these regions are not directly or indirectly involved in the binding of gp120 to CD4. PMID- 2548547 TI - [Gonadotropin, as a tumor marker, in body fluid and tumor tissues of germ cell tumors]. AB - The value of gonadotropin in the body fluids of germ cell tumor patients is its usefulness as a tumor marker. It is also used for differential diagnosis and/or judgement of therapeutic effects. In order to clarify the most effective value of gonadotropin as a tumor marker in the body fluids, we compared the value in serum, liquor and urine with one another. The liquor contained highest (1650 IU/l) value of gonadotropin in the primary intracranial germ cell tumors, mostly in choriocarcinoma. But the gonadotropin value was highest (3050 IU/l) in the serum of secondary intracranial choriocarcinoma. Chiasmal germ cell tumor, except choriocarcinoma, which does frequently secrete gonadotropin (alpha, beta) showed moderate or very high values in the liquor. However, pineal germ cell tumors rarely secrete gonadotropin and sometimes mild high value are obtained in the serum without gonadotropin secretion immunohistochemically. In such cases, the gonadotropin may be increased by indirect mechanism of gonadotropin-secretion following pineal disorder. In most of such cases, the gonadotropin was not human chorionic gonadotropin (HCG) but lutein hormone (LH). Because alpha-subunit of such gonadotropin has the same structure, their antibodies show immunologic cross reaction. So, a count of beta-subunit gonadotropin in the serum or liquor is the best way for differential diagnosis or judgement of therapeutic effects. From our results, it is considered that the tumor secretes HCG if the serum beta-HCG value was higher than 30 IU/l, and that it doesn't secrete HCG if beta-HCG value was lower than 10 IU/l or non calculable. The mild increased HCG may be caused by hypothalamo-diencephalic disorder such as pineal tumor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548548 TI - The variable effects of angiotensin converting enzyme inhibition on myocardial ischaemia in chronic stable angina. AB - The effect of angiotensin converting enzyme inhibition on myocardial ischaemia was studied in 12 normotensive patients with chronic stable angina and exercise induced ST segment depression. The study was randomised, double blind, placebo controlled, and crossover with treatment periods of two weeks. Enalapril was used to inhibit angiotensin converting enzyme. Assessment was by angina diaries and maximum symptom limited treadmill exercise tests. The results for the whole group showed a significant reduction in systolic blood pressure at rest and at peak exercise. Mean total exercise duration was 466 s (95% confidence interval 406 to 525) when the patients were taking placebo and 509 s (436 to 583) when they were taking enalapril. Four patients prolonged their total exercise time (mean 450 to mean 591 s) by more than 20%. Two patients, however, developed ischaemia earlier on exercise and reduced their total exercise duration (mean 490 to mean 390 s). Although angiotensin converting enzyme inhibition tended to reduce myocardial ischaemia in the group as a whole, some patients improved while others deteriorated. Thus the effects of enalapril are variable and this may have important implications when enalapril is used to treat heart failure in patients with underlying severe ischaemic heart disease. PMID- 2548549 TI - Automatic control of neuromuscular block with atracurium. AB - We have developed and tested a system for providing automatic control of neuromuscular block with atracurium. The degree of neuromuscular block was monitored in the small muscles of the hand using a Datex Relaxograph, and the signal was used as the controlled variable for a proportional plus integral controller. A preloaded integral was used to shorten the period of stabilization. The system was tested in 36 patients undergoing surgery and found to produce stable block. For a target T1 of 20% of control value, the mean block was 18.7 (SD 1.3) % of baseline T1 and the mean consumption of atracurium was 5.39 (SD 1.23) micrograms kg-1 min-1. The block was sufficiently stable to act as a background for studies of pharmacological and physiological interactions with atracurium. PMID- 2548550 TI - Chemical design of cilazapril. AB - 1. The three dimensional requirements for inhibition of ACE (angiotensin converting enzyme) were investigated in order to facilitate design of a more potent and selective antihypertensive agent. 2. All compounds designed possessed a bicyclic unit incorporating carboxylate and amidic carbonyl groups together with a thiol-bearing side chain. 3. NMR spectroscopy of the bicyclic units and molecular mechanics calculations enabled the possible positions of the thiol group to be studied. 4. Determination of the positions of the thiol group conferring best inhibition in the active site of ACE permitted the probable location of the active site zinc ion to be identified. The intention was to replace the thiol side chain with a homophenylalanine unit to bind to the zinc ion and also to occupy the S1 site which fits the Phe8 side chain of angiotensin I. 5. Examination of a torsional angle psi in a compound possessing poor inhibitory potency indicated correspondence to a high energy conformation of alanylproline. The bicyclic unit was modified to incorporate a seven-instead of a six-membered ring to bring psi into the range of an accessible conformation of alanylproline. The corresponding IC50 resulting indicated that psi was closer to that of the active conformations of enalaprilat and captopril. 6. Removal of one carbonyl improved the ACE inhibitory potency further. 7. The postulated active conformation of cilazaprilat is presented. PMID- 2548551 TI - The assessment of ACE activity in man following angiotensin I challenges: a comparison of cilazapril, captopril and enalapril. AB - 1. The aim of the studies was to develop a new methodology to estimate the pharmacodynamic properties and potency of angiotensin converting enzyme (ACE) inhibitors in man. 2. Angiotensin I dose-response curves were derived by continuous infusion of angiotensin I in increasing dose steps; steady state was reached within 3 min. 3. Interaction between angiotensin I (agonist) and ACE inhibitors (antagonist) was characterized according to Schild-plot methodology by measuring agonist dose-response curves using diastolic blood pressure in the absence and the presence of varying doses of the antagonist. 4. Cilazapril shifted the angiotensin I dose-response curves to the right. A twofold shift (apparent Ki-dose) was observed with approximately 0.6 mg cilazapril. 5. The effect of angiotensin I on diastolic blood pressure was determined before and up to 36 h after administration of 25 mg captopril, 10 mg enalapril, 4 mg cilazapril and a placebo orally. The pharmacodynamic half-life of captopril was about 2 h, whereas the effect of enalapril and cilazapril was about 4 h. 6. Angiotensin I dose-response curves are useful methods of investigating the pharmacodynamic properties of ACE-inhibitors in man. PMID- 2548552 TI - Effects of ACE inhibition with cilazapril on splanchnic and systemic haemodynamics in man. AB - 1. There is recent experimental evidence that the renin-angiotensin-system may play an essential role in producing splanchnic vasoconstriction. However, controversy exists as to the influence of ACE inhibition on splanchnic haemodynamics in man. We therefore investigated whether cilazapril, a structurally new and long-acting ACE inhibitor, interacts with angiotensin I dependent changes in splanchnic haemodynamics in man, using an experimental model. 2. The effects of cilazapril on angiotensin I-induced splanchnic and systemic haemodynamics were studied in seven normotensive men using the hepatic venous catheter technique (indocyanine-green dye), right-heart catheterisation (thermodilution method), intra-arterial blood pressure monitoring and systolic time-intervals. Dose-responses to angiotensin I were determined under control conditions and 60 min after ACE inhibition with 5 mg oral cilazapril. Angiotensin I was infused intravenously at constant rates in an increasing dose-sequence until systolic blood pressure was greater than 30 mm Hg. 3. ACE inhibition with cilazapril did not change basal splanchnic or systemic haemodynamics to any relevant extent. The angiotensin I dependent increase in systemic and pulmonary resistance and pulmonary capillary wedge pressure was attenuated by cilazapril, as indicated by the shift of the dose-response curves to the right. In the splanchnic vascular bed angiotensin I dose-dependently increased splanchnic vascular resistance and also wedge hepatic venous pressure and decreased splanchnic blood flow. These angiotensin I induced haemodynamic changes were clearly suppressed by cilazapril. The angiotensin I dose needed to produce a 30% increase in splanchnic vascular resistance, given as mean and s.e. mean, was 1.7 +/- 0.3 micrograms min-1 during control-trials vs 7.3 +/- 1.3 micrograms min-1 after ACE inhibition with cilazapril (P less than 0.001). 4. We conclude that, in man, the influence of cilazapril on acute angiotensin I-mediated haemodynamic responses is present in the splanchnic vascular bed, and that the overall effects of cilazapril are consistent with both arterial and venous effects of the ACE inhibitor. Cilazapril effectively counteracts angiotensin I-induced splanchnic vasoconstriction. PMID- 2548553 TI - The pharmacokinetics and angiotensin converting enzyme inhibition dynamics of cilazapril in essential hypertension. AB - 1. In a double-blind, placebo controlled, crossover study 12 patients with essential hypertension received single doses of 5, 10 and 20 mg of cilazapril. 2. Peak plasma levels of cilazaprilat and the 24 h areas under curve were directly proportional to dose. 3. The elimination half-life during the first 8 h was about 1.7 h. 4. From 24 h onwards there was a prolonged terminal elimination phase with a half-life of about 40 h, and strict dose proportionality of plasma concentrations was not maintained. 5. The pharmacokinetics of cilazaprilat and the pharmacodynamics of plasma ACE inhibition were well described by a one compartment model with saturable binding to ACE. 6. The coefficients of the model which related to plasma ACE and its interaction with cilazaprilat were in good agreement with model independent observations. 7. The values for kinetic and dynamic parameters in hypertensive patients were comparable with those reported for healthy volunteers. PMID- 2548554 TI - A pharmacokinetic study of cilazapril in patients with congestive heart failure. AB - 1. The pharmacokinetics of cilazapril and the inhibition of angiotensin converting enzyme (ACE) were investigated in 10 patients with congestive heart failure, NYHA class II-III, receiving diuretics with or without digoxin. 2. Patients received 0.5 mg and 1 mg cilazapril on the first 2 days, followed by 0.5 mg or 1 mg daily for the next 8 weeks, in a single-blind study. Plasma cilazaprilat concentrations and plasma ACE activities were measured by radioenzymatic methods up to 24 h after the first and last doses. 3. After the initial 0.5 mg dose of cilazapril, a mean maximum plasma concentration of cilazaprilat of 6.8 ng ml-1 was observed at 2.3 h. Concentrations declined up to 8 h with a mean half-life of 5.8 h, followed by slower decrease to 24 h. Total clearance, based on data to 24 h, was estimated at 8.5 l h-1, with three-fold inter-individual variation. Mean maximum plasma ACE inhibition was 87%, decreasing to 65% at 24 h. 4. In the multiple dose phase of the study, four patients received cilazapril 0.5 mg daily, and six patients 1 mg daily. Cilazapril accumulation for the 0.5 mg group averaged 77%, but steady state concentrations for the 1 mg group were less than double those of the 0.5 mg group. ACE inhibition profiles at steady state were similar for both groups, and they differed from first dose data only in a somewhat lower inhibition at 24 h. 5. Historical comparison of the first-dose data with those for healthy young volunteers at identical dosage revealed only minor differences in kinetic parameters.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548555 TI - Derivation of human microvascular endothelial cells for prosthetic vascular graft seeding. AB - This report summarizes our techniques and experiences deriving microvascular endothelial cells from fat for vascular graft seeding in 17 patients. Microvascular endothelial cells were derived from abdominal wall fat by collagenase incubation. The mean number of cells obtained using the described procedure was 6.83 x 10(5) cells/gram of fat processed. Histologic evaluation of the harvested cells revealed significant numbers of contaminating cell types in addition to Factor VIII-positive microvascular endothelial cells. These cells were seeded onto 6 mm ID PTFE vascular grafts in patients undergoing peripheral vascular arterial revascularization. The mean number of seeded cells was 8.04 x 10(6) cells/graft. Approximately 90 minutes were required to harvest and isolate the microvascular cells from the fat samples. We feel there are significant technical advantages to deriving endothelial cells from microvessels of human fat for vascular graft seeding. PMID- 2548556 TI - Detection of fibronectin on vascular flow surfaces by enzyme-linked immunosorbent assay. AB - Numerous variables are involved in the attachment of endothelial cells to a substrate. Quantifying these factors both on native vessels and on synthetic substrates is important in determining the success of endothelial cell attachment, retention, and growth on these substrates. Fibronectin is an important cell attachment molecule and is likely to be key to the successful attachment of endothelial cells to any substrate. For this reason we have developed an enzyme-linked immunosorbent assay for interrogation of the luminal surface of native and synthetic vessels for the presence of fibronectin. A plexiglass chamber was designed with two blocks, an upper block with wells and a lower supporting block. The chamber was then assembled with a vessel between the two blocks, forming the bottom of the well. This luminal surface was then interrogated by conventional enzyme-linked immunosorbent assay. Native vessels, collagenase-digested vessels, acellular matrices and PTFE preclotted with whole blood were assayed to determine the quantity of fibronectin present. These results were correlated with a bioassay developed to determine the quantity of fibronectin necessary for cell attachment. It was concluded that all of the samples assayed had ample fibronectin for cell attachment and that other factors must be responsible for successful maintenance of a cell monolayer. PMID- 2548557 TI - An improved procedure for enzymatic harvesting of highly purified canine venous endothelial cells for experimental small diameter vascular prostheses. AB - We developed a new device, the vein holder, to improve yield and purity of enzymatic harvests of venous endothelial cells. External jugular veins of mongrel dogs were dissected by a no-touch technique. In vitro length and circumference of the vein segments were decreased to about half of the in situ dimensions. The vein holder enabled mounting of the veins at 80% of their in situ length during endothelial cell harvesting. Trypan blue staining and scanning electron microscopic observations revealed that vein eversion as well as the new vein holder technique successfully removed the endothelium. Endothelial cell harvests by the eversion technique were, however, low and varied in size, viability, and purity. In contrast, the defined handling by the new vein holder technique regularly provided markedly increased amounts of endothelial cells. Most of the cells attached and developed cultures consisting of endothelial cells only, as shown by the uptake of DilAcLDL. Prostacyclin production of confluent cultures was similar to that of native veins. It is concluded that minimal handling, defined mounting, and prevention of overfilling the vein markedly improves endothelial cell harvests, providing greater amounts of viable and purified endothelial cells. PMID- 2548558 TI - A new cell line (8701-BC) from primary ductal infiltrating carcinoma of human breast. AB - A cell line, designated 8701-BC, was established in culture from tissue fragments of primary ductal infiltrating carcinoma of human breast. The cell cultures after the sixth passage were devoid of contaminating fibroblasts as judged by the positive staining of all cells with the specific epithelial cell markers carcinoembryonic antigen (CEA), tissue polypeptide antigen (TPA) and cytokeratin 8. The epithelial nature of these cells was confirmed by ultrastructural analyses which demonstrated the retention of specific structural properties characteristic of the original tumour. The cells possessed an abnormal karyotype with 55-60 chromosomes per cell with numerous rearrangements. They do not express HLA antigens and the c-myc gene was not amplified. The 8701-BC cells have a doubling time of approx. 29h and have been maintained in culture for more than 100 passages. These properties suggest the establishment of a human neoplastic cell line which, with its ability to produce homotrimer collagen in vitro, will provide a useful model system for the study of tumour cell:stromal matrix interactions. PMID- 2548559 TI - A case-control study of cervix cancer in Singapore. AB - Cervix cancer is about twice as common in Asia as in the Western world and its incidence varies among different Asian ethnic groups. A study based in Singapore, the population of which comprises Chinese, Indians and Malaysians, offers the opportunity to evaluate whether the same risk factors are important in this part of the world as in the West. A total of 135 cases and an equal number of controls were interviewed and details concerning reproductive and sexual history, smoking, hygiene, socio-economic status and education were collected. Seventy-three cases had invasive cancer while 62 had micro-invasive disease or CIN III. The most important risk factors were parity and number of sexual partners. Smoking was rare in cases and controls and did not appear to be an important determinant of risk. Of the socio-economic factors, education appeared most predictive and lowered the risk. Age at first intercourse was strongly correlated with education (positively) and parity (negatively), but not with number of sexual partners. Biopsies were available for HPV DNA analysis in 38 cases and 37% were positive, mostly for HPV type 16. All these factors gave similar risks in invasive and preinvasive disease. PMID- 2548560 TI - A simple outpatient treatment with oral ifosfamide and oral etoposide for patients with small cell lung cancer (SCLC). AB - For the first time in a clinical study oral Ifosfamide was used: 65 elderly or unfit patients with small cell lung cancer (SCLC) were treated as outpatients with fractionated oral Ifosfamide and Etoposide. Forty patients (62%) had extensive stage (ED) disease. The median age of the patients was 66 years. In the 60 patients evaluable for response the objective response rate was 90% with a complete response (CR) rate of 32% and a partial response (PR) rate of 58%. The overall median survival of all 65 patients was 11 months (13 months for LD, 9.5 months for ED). In those patients with LD achieving a CR or a PR radiotherapy was given to the mediastinum. No prophylactic cranial irradiation was given. There was a rapid improvement in the responding patients' performance status and symptoms generally with the first treatment cycle. Overall haematological toxicity was mild, with intravenous antibiotics only being required in 4% of the courses and with only one treatment-related death from septicaemia. A higher than expected rate of CNS toxicity was seen (30%). This was generally mild and always fully reversible and consisted mainly of forgetfulness, occasionally hallucinations, nightmares and somnolence. In only one case did encephalopathy necessitate early termination of treatment. This raises the question of whether Ifosfamide metabolism differs quantitatively or qualitatively when given by the oral route as opposed to the usual intravenous route. We conclude that this simple outpatient based treatment gives a high response rate with rapid improvement in symptoms. PMID- 2548561 TI - Identification of immunoglobulin-containing cells in the central nervous system of the mouse following infection with the demyelinating strain of Semliki Forest virus. AB - Cells within the central nervous system were identified as containing immunoglobulin G, A and M using immunocytochemistry in mice previously infected with Semliki Forest virus, a togavirus causing primary immune-mediated demyelination. Cells positive for these immunoglobulins were counted in cerebellar white matter, parenchyma, meninges and choroid plexus/ventricles. No positively staining cells were seen on day 6 after infection although other inflammatory cells were present at this time and virus-specific immunoglobulin was found in serum. Cells positive for IgG appeared in all areas by day 9 and remained dominant in numbers throughout. IgM-secreting cells appeared in small numbers in the parenchyma first on day 9 and subsequently in other areas, their numbers rising to a maximum on day 12 in all areas and falling thereafter. The number of IgA-secreting cells was small. They appeared by PID 12 and continued to rise on successive sampling days. Initially IgG-positive cells were seen in the perivascular cuffs but by day 12 a few had moved away from the cuffs into the adjacent parenchyma. IgG-positive cells were seen both in and away from cuffs within areas of demyelination. IgM and IgA-positive cells tended to follow the distribution of IgG-positive cells, but in fewer numbers. PMID- 2548562 TI - Coxsackievirus B3-induced acute pancreatitis: analysis of histopathological and viral parameters in a mouse model. AB - Coxsackievirus B3 infection in mice was studied histopathologically, by virus isolation and by nucleic acid hybridization after intraperitoneal inoculation of the virus. Extensive viraemia was detected for 1-3 days post-infection. All mice developed necrotizing acute pancreatitis and focal myocarditis. Pancreatitis eventually lead to complete atrophy of the exocrine pancreas. However, the islets of Langerhans and pancreatic ducts remained morphologically intact. Virus could be demonstrated in pancreatic tissue for 1-5 days post-infection by in-situ and spot hybridization as well as by virus isolation. Virus was not detectable on days 7-22 post-infection suggesting an autodigestive aetiology in further destruction of the exocrine pancreas. The mouse model described here permits detailed analysis of viral and host factors in the pathogenesis of enterovirus infections. Since coxsackie B viruses have been proposed to be aetiological agents in human acute pancreatitis, the application of in-situ hybridization allows analysis of enteroviruses directly from pancreatic tissue of clinical routine specimens. PMID- 2548563 TI - Correlation of membrane anisotropy with function in subpopulations of human blood platelets. AB - Human blood platelets were fractionated on a discontinuous Percoll gradient into high density (HD), intermediate (ID), and low density (LD) platelets. The subpopulations were characterized with regard to [14C]serotonin uptake and release, cAMP content, aggregation, and membrane anisotropy, which is inversely related to membrane fluidity. Membrane anisotropy, which was high in LD platelets, was found to decrease with increasing density (LD greater than ID greater than HD). LD platelets showed significant lower cAMP levels and [14C]serotonin uptake than the total platelet population (TPP) and ID and HD platelets. Upon ADP and serotonin stimulation the cAMP content was reduced in all platelet populations with the exception of HD platelets in which cAMP was unchanged. Upon thrombin stimulation the cAMP content was reduced only in TPP and LD platelet population and it was increased in HD platelet population. Thrombin activation changed the anisotropy only in LD platelets. Thrombin at a concentration of 0.001 U/ml reduced whereas 0.01 and 0.05 U thrombin/ml increased the membrane anisotropy significantly. As compared with TPP and the other subpopulations, LD platelets were most sensitive upon ADP and thrombin stimulated [14C]serotonin release as well as upon ADP, serotonin and thrombin induced aggregation. The findings suggest that the differing functional abilities of the platelet subpopulations are correlated to the various membrane anisotropies observed in these fractions. PMID- 2548564 TI - Natural history of pleural thickening after exposure to crocidolite. AB - Serial plain chest radiographs of 384 men who worked at the Wittenoom crocidolite mine and mill between 1943 and 1966 and who applied for pneumoconiosis compensation between 1948 and 1982 have been examined independently by two trained observers for pleural disease using the 1980 ILO-UICC classification of radiographs to record width and extent of pleural disease. Radiographs covering follow up periods of from two to 38 years were examined (median number of films per subject was nine). The degree of crocidolite exposure was estimated from employment records and a survey of airborne fibre concentrations performed in 1966. Agreement between the observers on the presence and degree of pleural disease in the final film for each subject was moderately close (Kendall's tau B = 0.62) and was least for subjects with thickening less than 5 mm in width. Diffuse pleural thickening extending for greater than 50% of the lateral chest wall was the most common type recorded by both observers. Minor pleural thickening frequently progressed in extent along the lateral chest wall but progression beyond 5 mm in thickness was less common. Pleural plaques were not seen to progress beyond their initial thickness or extent. The rate of onset of thickening in this population increased continually from the time of first exposure and also increased slightly with age. There was evidence that the level of total cumulative exposure to crocidolite increased the rate of onset of pleural thickening in the period between five and 15 years after first exposure. Rate of progression of established thickening was greatest in subjects who first developed thickening early after first exposure. The relative rate of progression decreased slowly with time from first signs of thickening and there was no evidence of any progression more than 15 years after onset. PMID- 2548565 TI - Effect of retinoic acid on asbestos induced plasminogen activator activity of peritoneal macrophages. PMID- 2548566 TI - What threat is human parvovirus B19 to the fetus? A review. PMID- 2548567 TI - Fetal pathology in human parvovirus B19 infection. AB - In a current Netherlands study on the effects on mother and child of infection with the human parvovirus B19 during pregnancy, 10 pregnancies have been reported. Three of them ended before term: two in fetal death and one by elective abortion. In two of these fetuses B19 infection in cells other than those of the erythroid series was demonstrated, and in the one terminated, ocular malformation and extensive inflammatory reactions in all fetal and placental tissues were found. The presence of B19 DNA was demonstrated by dot hybridization in placental and fetal tissues. In the third no gross fetal abnormalities were found, although B19 DNA was detected in several fetal tissues by in-situ hybridization. Of the remaining seven pregnancies, six ended at term in the birth of apparently healthy babies. The other child was born near term with a low birthweight and multiple congenital malformations, but with no proof of intrauterine B19 infection. It is concluded that B19 infection in pregnancy can interfere with organ development and may lead to intrauterine fetal death. PMID- 2548568 TI - The role of low-dose methotrexate and folinic acid in gestational trophoblastic tumours (GTT). AB - Between 1964 and 1986, 487 patients with gestational trophoblastic tumour (GTT) were treated with methotrexate and folinic acid. The patients comprise two groups: between 1964 and 1974, 126 patients were treated but were not systematically stratified using a prognostic score before the start of treatment. These patients formed part of the 317 women who were analysed to identify a number of prognostic variables (Bagshawe 1976). Retrospective analysis of these 126 patients using these prognostic factors showed that in the true low-risk group 85/88 (96%) are alive while 20/22 (91%) of the medium-risk group and only 5/16 (31%) of the high-risk group are alive. Overall the survival was 110/126 (87%) with a minimum follow-up of 14 years. From 1974 all patients were stratified on admission into prognostic groups. Of the true low-risk patients 347/348 survived (99.7%); 13 patients were underscored and treated as low risk when they should have been treated as medium risk, 12 (92%) of these are alive, but nine (69%) needed to change treatment because of drug resistance. While the overall survival in the 1974-1986 group was 359/361 (99%) with a minimum follow up of 16 months, the survival in all patients (1964-1986) was 469/487 (96%). Although the survival in these patients is excellent it should be noted that 69/348 (20%) low-risk patients had to change treatment because of the development of drug resistance, and a further 23 (6%) needed to change treatment because of drug-induced toxicity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548569 TI - pH dependence of the kinetic parameters for the pyrophosphate-dependent phosphofructokinase reaction supports a proton-shuttle mechanism. AB - The pH dependence of kinetic parameters for the pyrophosphate-dependent phosphofructokinase from Propionibacterium freudenreichii suggests that the enzyme catalyzes its reaction via general acid-base catalysis with the use of a proton shuttle. The base is required unprotonated in both reaction directions. In the direction of fructose 6-phosphate phosphorylation the base accepts a proton from the hydroxyl at C-1 of F6P and then donates it to protonate the leaving phosphate. Whether this occurs in one or two steps cannot be deduced from the present data. The maximum velocity of the reaction is pH independent in both reaction directions while V/K profiles exhibit pKs for binding groups (including enzyme and reactant functional groups) as well as pKs for enzyme catalytic groups. These data suggest that reactants bind only when correctly protonated and only to the correctly protonated form of the enzyme. Specifically, the requirement for two enzyme epsilon-amino groups in the protonated form for reactant binding was detected as was the requirement for the ionized phosphates of fructose 6-phosphate, fructose 1,6-bisphosphate, MgPPi and HPO4(2-). The protonation state of enzyme and reactant binding groups is in agreement with data obtained previously [Cho, Y.-K., & Cook, P. F. (1988) J. Biol. Chem. 263, 5135]. PMID- 2548570 TI - Rapid isolation of OmpF porin-LPS complexes suitable for structure-function studies. AB - A gentle and rapid isolation procedure is described yielding fractions containing better than 95% pure OmpF porin of Escherichia coli BE with different amounts of bound lipopolysaccharide (LPS). The procedure employs continuous free-flow electrophoresis (FFE) in the presence of detergent above its critical micelle concentration. Total yields of around 45% were typically obtained when porin enriched membrane extracts were processed. By use of analytical ultracentrifugation a molecular mass of 114,000 and a sedimentation coefficient S20,w of 5.0 S were determined for porin trimers containing approximately 1 mol of tightly bound LPS. This porin readily formed 3D crystals suitable for high resolution X-ray diffraction analysis. Three other porin-LPS isoforms isolated by FFE revealed molecular masses of 120,000, 124,000, and 151,000, suggesting that, in addition to the tightly bound LPS, 1, 2, and 8 mol of loosely bound LPS were present per mole of porin trimer. Each of the four different isoforms was suitable for reconstitution into highly ordered protein-lipid membrane arrays. The membrane crystals obtained with the 151-kDa isoform exhibited a unit cell polymorphism similar to that previously reported. PMID- 2548571 TI - Kinetics of [3H]muscimol binding to the GABAA receptor in bovine brain membranes. AB - The binding of the GABA receptor agonist [3H]muscimol to membrane preparations from bovine cerebral cortex has been investigated in equilibrium and kinetic experiments. Equilibrium binding curves are biphasic and suggest that [3H]muscimol binds to both high-affinity (Kd approximately 10 nM) and low affinity (Kd approximately 0.5 microM) sites. Binding to each class of sites is inhibited by GABA and by the specific GABAA receptor antagonist bicuculline. The kinetics of [3H]muscimol binding have been measured by using both manual filtration assays and an automated rapid filtration technique which permits the measurement of ligand dissociation on subsecond time scales. Association and dissociation curves are biphasic at all concentrations of [3H]muscimol studied, even under conditions of low receptor saturation when no significant occupancy of the low-affinity sites would be expected. These results cannot be simply explained by the presence of two populations of binding sites in the membrane preparations but suggest the existence of two forms of the monoliganded receptor. Dissociation constants for these two forms have been estimated to be 16 and 82 nM at 23 degrees C. At higher ligand concentrations, kinetic measurements have suggested that the binding of [3H]muscimol to low-affinity sites is accompanied by a slow conformational change of the receptor-ligand complex. PMID- 2548572 TI - Primary structure of the cAMP-dependent phosphorylation site of the plasma membrane calcium pump. AB - The primary structure of a region of the erythrocyte plasma membrane calcium pump which is phosphorylated by the cAMP-dependent protein kinase has been determined. The sequence is A-P-T-K-R-N-S-S(P)-P-P-P-S-P-D. The site is located between the calmodulin binding domain and the C-terminus of the ATPase. The ATPase is phosphorylated only at this site by the cAMP-dependent protein kinase, and the phosphorylation is inhibited by calmodulin. The effect of the phosphorylation is to decrease the Km for Ca2+ of the purified ATPase from about 10 microM to about 1.4 microM and to increase the Vmax of ATP hydrolysis about 2-fold. PMID- 2548573 TI - Static and transient hydrogen-bonding interactions in recombinant desulfatohirudin studied by 1H nuclear magnetic resonance measurements of amide proton exchange rates and pH-dependent chemical shifts. AB - With proton nuclear magnetic resonance spectroscopy at 22 degrees C and pD 4.5, individual exchange rates in the range from 2 X 10(-5) to 1 X 10(-1) min-1 were observed for 23 amide protons in recombinant desulfatohirudin. The remaining 38 backbone amide protons exchange more rapidly than 1 X 10(-1) min-1. All 23 slowly exchanging protons are located in the polypeptide segment from residue 4 to residue 42, which forms a well-defined globular domain. Three different breathing modes of this molecular region are manifested in the exchange data, which appear to be correlated with the location of the three disulfide bonds. Chemical shift changes larger than 0.15 ppm between pH 2.5 and pH 5.0 arising from through-space interactions with carboxyl groups were observed for seven backbone amide protons. Two of these shifts can be explained by hydrogen bonds in the core of the protein, Gly 25 NH-Glu 43 O epsilon and Ser 32 NH-Asp 33 O delta, and two others by intraresidual NH-O epsilon interactions in Glu 61 and Glu 62. The remaining three pH shifts for Glu 35, Cys 39, and Ile 59 imply the existence of transient interactions between the molecular core and the flexible C-terminal segment 49 65, which have so far not been characterized by nuclear Overhauser effects or other conformational constraints. PMID- 2548574 TI - Nucleotide and AP5A complexes of porcine adenylate kinase: A 1H and 19F NMR study. AB - Proton and fluorine nuclear magnetic resonance spectroscopies (NMR) were used as methods to investigate binary complexes between porcine adenylate kinase (AK1) and its substrates. We also studied the interaction of fluorinated substrate analogues and the supposed bisubstrate analogue P1,P5-bis(5'-adenosyl) pentaphosphate (AP5A) with AK1 in the presence of Mg2+. The chemical shifts of the C8-H, C2-H, and ribose C1'-H resonances of both adenosine units in stoichiometric complexes of AK1 with AP5A in the presence of Mg2+ could be determined. The C2-H resonance of one of the adenine bases experiences a downfield shift of about 0.8 ppm on binding to the enzyme. The chemical shift of the His36 imidazole C2-H was changed in the downfield direction on ATP-Mg2+ and, to a lesser extent, AMP binding. 19F NMR chemical shifts of 9-(3-fluoro-3-deoxy beta-D-xylofuranosyl)adenine triphosphate (3'-F-X-ATP)-Mg2+ and 9-(3-fluoro-3 deoxy-beta-D-xylofuranosyl)adenine monophosphate (3'-F-X-AMP) bound to porcine adenylate kinase could be determined. The different chemical shifts of the bound nucleotides suggest that their mode of binding is different. Free and bound 3'-F X-AMP are in fast exchange with respect to their 19F chemical shifts, whereas free and bound 3'-F-X-ATP are in slow exchange on the NMR time scale in the absence as well as in the presence of Mg2+. This information could be used to determine the apparent dissociation constants of the nucleotides and the 3'-F-X analogues in the binary complexes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548575 TI - 14 beta-(Bromoacetamido)morphine irreversibly labels mu opioid receptors in rat brain membranes. AB - The binding properties of 14 beta-(bromoacetamido)morphine (BAM) and the ability of BAM to irreversibly inhibit opioid binding to rat brain membranes were examined to characterize the affinity and selectivity of BAM as an irreversible affinity ligand for opioid receptors. BAM had the same receptor selectivity as morphine, with a 3-5-fold decrease in affinity for the different types of opioid receptors. When brain membranes were incubated with BAM, followed by extensive washing, opioid binding was restored to control levels. However, when membranes were incubated with dithiothreitol (DTT), followed by BAM, and subsequently washed, 90% of the 0.25 nM [3H] [D-Ala2,(Me)Phe4,Gly(ol)5]enkephalin (DAGO) binding was irreversibly inhibited as a result of the specific alkylation of a sulfhydryl group at the mu binding site. This inhibition was dependent on the concentrations of both DTT and BAM. The mu receptor specificity of BAM alkylation was demonstrated by the ability of BAM alkylated membranes to still bind the delta-selective peptide [3H] [D-penicillamine2,D-penicillamine5]enkephalin (DPDPE) and (-)-[3H]bremazocine in the presence of mu and delta blockers, selective for kappa binding sites. Under conditions where 90% of the 0.25 nM [3H]DAGO binding sites were blocked, 80% of the 0.8 nM [3H]naloxone binding and 50% of the 0.25 nM 125I-labeled beta h-endorphin binding were inhibited by BAM alkylation. Morphine and naloxone partially protected the binding site from alkylation with BAM, while ligands that did not bind to the mu site did not afford protection.2+hese studies have demonstrated that when a disulfide bond PMID- 2548577 TI - Endonuclease III is an iron-sulfur protein. AB - Elemental analyses, Mossbauer, and EPR data are reported to show that endonuclease III of Escherichia coli is an iron-sulfur protein. Mossbauer spectra of protein freshly prepared from E. coli grown on 57Fe-enriched medium demonstrate that the native enzyme contains a single 4Fe-4S cluster in the 2+ oxidation state, with a net spin of zero. Upon treatment with ferricyanide, a fraction (less than 25%) of the clusters is oxidized into a state which yields an EPR spectrum near g = 2.01 typical of a 3Fe-4S cluster. The magnetic field dependence of the linear electric field effect verifies this assignment. Electron spin echo modulation on the g = 2.01 form of the protein in deuterated solvent indicates the presence of exchangeable protons in the vicinity of the 3Fe-4S cluster. The data obtained show that the [4Fe-4S]2+ cluster of the native enzyme is resistant to either oxidation or reduction, although photoreduction elicited a g = 1.94 type EPR signal characteristic of a [4Fe-4S]1+ cluster. These studies show that endonuclease III is unique in being both a DNA repair enzyme and an iron-sulfur protein. The function of the 4Fe-4S cluster remains to be established. PMID- 2548576 TI - Kinetic properties of the Na+/H+ antiport of heart mitochondria. AB - The fluorescence of 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) has been used to follow the Na+/H+ antiport activity of isolated heart mitochondria as a Na+-dependent extrusion of matrix H+. The antiport activity measured in this way shows a hyperbolic dependence on external Na+ or Li+ concentration when the external pH (pHo) is 7.2 or higher. The apparent Km for Na+ decreases with increasing pHo to a limit of 4.6 mM. The Ki for external H+ as a competitive inhibitor of Na+/H+ antiport averages 3.0 nM (pHo 8.6). The Vmax at 24 degrees C is 160 ng ion of H+ min-1 (mg of protein)-1 and does not vary with pHo. Li+ reacts with the antiporter with higher affinity, but much lower Vmax, and is a competitive inhibitor of Na+/H+ antiport. The rate of Na+/H+ antiport is optimal when the pHi is near 7.2. When pHo is maintained constant, Na+-dependent extrusion of matrix H+ shows a hyperbolic dependence on [H+]i with an apparent Km corresponding to a pHi of 6.8. The Na+/H+ antiport is inhibited by benzamil and by 5-N-substituted amiloride analogues with I50 values in the range from 50 to 100 microM. The pH profile for this inhibition seems consistent with the availability of a matrix binding site for the amiloride analogues. The mitochondrial Na+/H+ antiport resembles the antiport found in the plasma membrane of mammalian cells in that Na+, Li+, and external H+ appear to compete for a common external binding site and both exchanges are inhibited by amiloride analogues.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548578 TI - Comparison of the data, analysis, and results of X-ray absorption studies of cytochrome c oxidase. AB - Differences in the methods of analysis of X-ray absorption data used by Powers et al. [Powers, L., Blumberg, W. E., Chance, B., Barlow, C., Leigh, J., Jr., Smith, J., Yonetani, T., Vik, S., & Peisach, J. (1979) Biochim. Biophys. Acta 547, 520 538; Powers, L., Chance, B., Ching, Y., & Angiolillo, P. (1981) Biophys. J. 34, 465-498] and Scott et al. [Scott, R., Schwartz, J., & Cramer S. (1986) Biochemistry 25, 5546-5555] are clarified. In addition, we compare the X-ray absorption data and results for resting cytochrome c oxidase reported by both groups using the same analysis method and conclude apart from any assumptions that the data are not identical. PMID- 2548579 TI - Structure of the human thyroid peroxidase gene: comparison and relationship to the human myeloperoxidase gene. AB - All exons of the human thyroid peroxidase gene were cloned from phage and cosmid libraries and sequenced, including 2599 base pairs of upstream DNA. The gene contains 17 exons and covers at least 150 kilobase pairs of chromosome 2. The transcription start site was identified by both S1 mapping and primer extension; a typical TATA box was found 25 bases upstream of the putative start site. A comparison of the gene structures of thyroid peroxidase and a granulocyte protein, myeloperoxidase, revealed that the positions of the 3rd through 11th exon-intron junctions in thyroid peroxidase coincide exactly with those of the 2nd through 11th exon-intron junctions in myeloperoxidase except the 7th myeloperoxidase junction, that does not have any counterpart in thyroid peroxidase. The amino acid codon separation pattern in each junction is well conserved between both enzymes. Four exons, unique to thyroid peroxidase, are located at the 3' end of the gene (exons 13-16), each of which encompasses a different protein module. Three of these modules, representing exons 13, 14, and 15, bear significant similarities to C4b-beta 2 glycoprotein, the EGF-LDL receptor, and a typical transmembrane domain, respectively. The genes coding for these modules were probably fused to an ancestral peroxidase gene to generate the present thyroid peroxidase gene. The data suggest that intron loss, and/or insertion, and exon shuffling have played important roles in the evolution of the thyroid peroxidase gene. PMID- 2548581 TI - Mutation of glycine 49 to valine in the alpha subunit of GS results in the constitutive elevation of cyclic AMP synthesis. AB - The G-protein GS couples hormone-activated receptors with adenylyl cyclase and stimulates increased cyclic AMP synthesis. Transient expression in COS-1 cells of cDNAs coding for the GS alpha-subunit (alpha S) or alpha S cDNAs having single amino acid mutations Gly49----Val or Gly225----Thr elevated cyclic AMP levels, resulting in the activation of cyclic AMP dependent protein kinase. Stable expression in Chinese hamster ovary cells of alpha S Val49 cDNA resulted in a small constitutive elevation of cyclic AMP that was sufficient to persistently activate cyclic AMP dependent protein kinase activity 1.5-2-fold over basal activity. Stable expression of wild-type alpha S or alpha S Thr225 in Chinese hamster ovary cells was less effective in sustaining elevated cyclic AMP synthesis and kinase activation compared to alpha SVal49. PMID- 2548580 TI - Antisera specific for the alpha 1, alpha 2, alpha 3, and beta subunits of the Na,K-ATPase: differential expression of alpha and beta subunits in rat tissue membranes. AB - We have developed a panel of antibodies specific for the alpha 1, alpha 2, alpha 3, and beta subunits of the rat Na,K-ATPase. TrpE-alpha subunit isoform fusion proteins were used to generate three antisera, each of which reacted specifically with a distinct alpha subunit isotype. Western blot analysis of rat tissue microsomes revealed that alpha 1 subunits were expressed in all tissues while alpha 2 subunits were expressed in brain, heart, and lung. The alpha 3 subunit, a protein whose existence had been inferred from cDNA cloning, was expressed primarily in brain and copurified with ouabain-inhibitable Na,K-ATPase activity. An antiserum specific for the rat Na,K-ATPase beta subunit was generated from a TrpE-beta subunit fusion protein. Western blot analysis showed that beta subunits were present in kidney, brain, and heart. However, no beta subunits were detected in liver, lung, spleen, thymus, or lactating mammary gland. The distinct tissue distributions of alpha and beta subunits suggest that different members of the Na,K-ATPase family may have specialized functions. PMID- 2548582 TI - Crystal structure of a cAMP-independent form of catabolite gene activator protein with adenosine substituted in one of two cAMP-binding sites. AB - Catabolite gene activator protein (CAP) in the presence of cAMP stimulates transcription from several operons in Escherichia coli. A cAMP-independent variant, in which Ala-144 is replaced by Thr (CAP91), is activated by analogues of cAMP, such as adenosine, which do not activate the wild-type CAP. In order to test the effect of adenosine on the structure, a crystal of CAP91 grown as a complex with cAMP was soaked in a solution of 10 mM adenosine, and X-ray diffraction data were measured to 3.5-A resolution. The difference Fourier map calculated with phases from the CAP91 structure showed significant negative density at the position of the phosphate of cAMP bound in one subunit of the CAP91 dimer. Adenosine was preferentially substituted for cAMP in the subunit in the "closed" conformation, while the cAMP-binding site of the "open" subunit was apparently still occupied by cAMP. The structure was refined by restrained least squares methods to an R factor of 20.2%. Adenosine is not bound in exactly the same position as cAMP; instead, the 5'-OH of adenosine is in a new position that allows formation of two hydrogen bonds with Ser-83, replacing two of the three interactions of the phosphate of cAMP with Arg-82 and Ser-83. PMID- 2548583 TI - A novel primase-free form of murine DNA polymerase alpha induced by infection with minute virus of mice. AB - Two species of DNA polymerase alpha free of primase activity were identified in extracts of Ehrlich mouse cells that had been infected with minute virus of mice. Primase-free forms of DNA polymerase alpha eluted with 150 and 180 mM NaCl during ion-exchange chromatography on DEAE-cellulose columns, exhibited sedimentation coefficients of 11 S and 8.2 S, respectively, and were inhibited by aphidicolin, N2-(p-n-butylphenyl)-9-(2-deoxy-beta-D-ribofuranosyl)guanine 5'-triphosphate, and 2-(p-n-butylanilino)-9-(2-deoxy-beta-D-ribofuranosyl)adenine 5'-triphosphate. The ratio of primase-free DNA polymerase alpha to the DNA polymerase alpha-primase complex increased from 1.5 to greater than 100 during the course of infection, and free primase was produced during the MVM replicative cycle. PMID- 2548584 TI - On the mechanism of topoisomerase I inhibition by camptothecin: evidence for binding to an enzyme-DNA complex. AB - Camptothecin, a cytotoxic antitumor compound, has been shown to produce protein linked DNA breaks mediated by mammalian topoisomerase I. We have investigated the mechanism by which camptothecin disrupts DNA processing by topoisomerase I and have examined the effect of certain structurally related compounds on the formation of a DNA-topoisomerase I covalent complex. Enzyme-mediated cleavage of supercoiled plasmid DNA in the presence of camptothecin was completely reversed upon the addition of exogenous linear DNA or upon dilution of the reaction mixture. Camptothecin and topoisomerase I produced the same amount of cleavage from supercoiled DNA or relaxed DNA. In addition, the alkaloid decreased the initial velocity of supercoiled DNA relaxation mediated by catalytic quantities of topoisomerase I. Inhibition occurred under conditions favoring processive catalysis as well as under conditions favoring distributive catalysis. By use of [3H]camptothecin and an equilibrium dialysis assay, the alkaloid was shown to bind reversibly to a DNA-topoisomerase I complex, but not to isolated enzyme or isolated DNA. These results are consistent with a model in which camptothecin reversibly traps an intermediate involved in DNA unwinding by topoisomerase I and thereby perturbs a set of equilibria, resulting in increased DNA cleavage. By examining certain compounds that are structurally related to camptothecin, it was found that the 20-hydroxy group, which has been shown to be essential for antitumor activity, was also necessary for stabilization of the covalent complex between DNA and topoisomerase I. In contrast, no such correlation existed for UV light-induced cleavage of DNA by Cu(II)-camptothecin derivatives. PMID- 2548585 TI - Structure-activity relationships in the hydrolysis of substrates by the phosphotriesterase from Pseudomonas diminuta. AB - The mechanism and substrate specificity of the phosphotriesterase from Pseudomonas diminuta have been examined. The enzyme hydrolyzes a large number of phosphotriester substrates in addition to paraoxon (diethyl p-nitrophenyl phosphate) and its thiophosphate analogue, parathion. The two ethyl groups in paraoxon can be changed to propyl and butyl groups, but the maximal velocity and Km values decrease substantially. The enzyme will not hydrolyze phosphomonoesters or -diesters. There is a linear correlation between enzymatic activity and the pKa of the phenolic leaving group for 16 paraoxon analogues. The beta value in the corresponding Bronsted plot is -0.8. No effect on either Vmax or Vmax/Km is observed when sucrose is used to increase the relative solvent viscosity by 3 fold. These results are consistent with rate-limiting phosphorus-oxygen bond cleavage. A plot of log V versus pH for the hydrolysis of paraoxon shows one enzymatic group that must be unprotonated for activity with a pKa of 6.1. The deuterium isotope effect by D2O on Vmax and Vmax/Km is 2.4 and 1.2, respectively, and the proton inventory is linear, which indicates that only one proton is "in flight" during the transition state. The inhibition patterns by the products are consistent with a random kinetic mechanism. PMID- 2548586 TI - Rapid freeze- and chemical-quench studies of dopamine beta-monooxygenase: comparison of pre-steady-state and steady-state parameters. AB - The copper-containing enzyme dopamine beta-monooxygenase has been studied with regard to pre-steady-state kinetics of tyramine hydroxylation and reduction of enzyme-bound Cu2+ by chemical- and freeze-quench EPR techniques. The bulk of the enzyme-bound copper (approximately 70%) is reduced in a single-exponential process with a limiting rate constant of 250 s-1, Km = 0.9 mM, consistent with participation of both copper ions in the redox events of catalysis. The remaining copper is reduced much more slowly (k approximately 2 s-1) or not at all, attributed to a distribution of copper into inhibitory binding sites and the presence of some inactive enzyme. Knowledge of the Cu2+ reduction rate, together with rate constants calculated from steady-state isotope effects [Miller, S. M., & Klinman, J. P. (1985) Biochemistry 24, 2114-2127], has allowed prediction of pre-steady-state product formation transients. Measurement of these transients under conditions of excess ascorbate shows close agreement with prediction, supporting the validity of individual rate constants obtained from steady-state data. Kinetic modeling shows further that the predominant steady-state enzyme form is the enzyme-product complex (E-P), which is expected to show a correspondingly large (approximately 70% of total copper) EPR signal for bound Cu2+. Surprisingly, the steady state is characterized by a low (19% of total copper) EPR signal. This lack of correlation between the anticipated and observed steady-state EPR signal suggests either antiferromagnetic coupling in binuclear copper centers or reduction of Cu2+ in this enzyme form by ascorbic acid.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548587 TI - Correlation of copper valency with product formation in single turnovers of dopamine beta-monooxygenase. AB - Chemical- and freeze-quench EPR techniques have allowed single-turnover studies of the copper-containing enzyme dopamine beta-monooxygenase. Reduction of enzyme by a stoichiometric amount of ascorbate followed by rapid mixing with tyramine leads to oxidation of bound copper and formation of hydroxylated product in the expected 2:1 ratio. The tyramine dependence of single turnovers yields a limiting rate of 82 +/- 9 s-1 and Km of 3 +/- 1 mM, in agreement with kinetic modeling based on steady-state parameters. Together these results show that the reduced enzyme is a catalytically competent species, with bound copper acting as the sole reservoir of reducing equivalents. The correlation of copper oxidation and substrate hydroxylation rules out significant antiferromagnetic spin coupling in the enzyme-product complex. Since the enzyme-product complex's Cu2+ EPR signal is absent in the transient approach to the steady state [Brenner, M. C., Murray, C. J., & Klinman, J. P. (1989) Biochemistry (preceding paper in this issue)], this result implies that ascorbate reduces copper in the enzyme-product complex. These findings have important consequences for catalysis and active site structure in dopamine beta-monooxygenase. PMID- 2548588 TI - Kinetic studies of the pyridoxal kinase from pig liver: slow-binding inhibition by adenosine tetraphosphopyridoxal. AB - Pyridoxal kinase from pig liver has been purified 10,000-fold to apparent homogeneity. The enzyme is a dimer of subunits of Mr 32,000. The enzyme is strongly inhibited by the product pyridoxal 5'-phosphate. Liver pyridoxamine phosphate oxidase, another enzyme involved in the biosynthesis of pyridoxal 5' phosphate, is also strongly inhibited by this compound [Wada, H., & Snell, E. E. (1961) J. Biol. Chem. 236, 2089-2095]. Thus, the biosynthesis of pyridoxal 5' phosphate in the liver might be regulated by the product inhibition of both pyridoxamine phosphate oxidase and pyridoxal kinase. Kinetic studies revealed that the catalytic reaction of liver pyridoxal kinase follows an ordered mechanism in which pyridoxal and ATP bind to the enzyme and ADP and pyridoxal 5' phosphate are released from the enzyme, in this order. Adenosine tetraphosphopyridoxal was found to be a slow-binding inhibitor of pyridoxal kinase. Pre-steady-state kinetics of the inhibition revealed that the inhibitor and the enzyme form an initial weak complex prior to the formation of a tighter and slowly reversing complex. The overall inhibition constant was 2.4 microM. ATP markedly protects the enzyme against time-dependent inhibition by the inhibitor, whereas another substrate pyridoxal affords no protection. By contrast, adenosine triphosphopyridoxal is not a slow-binding inhibitor of this enzyme. PMID- 2548589 TI - Binding of thiocyanate to lactoperoxidase: 1H and 15N nuclear magnetic resonance studies. AB - The binding of thiocyanate to lactoperoxidase (LPO) has been investigated by 1H and 15N NMR spectroscopy. 1H NMR of LPO shows that the major broad heme methyl proton resonance at about 61 ppm is shifted upfield by addition of the thiocyanate, indicating binding of the thiocyanate to the enzyme. The pH dependence of line width of 15N resonance of SC15N- in the presence of the enzyme has revealed that the binding of the thiocyanate to the enzyme is facilitated by protonation of an ionizable group (with pKa of 6.4), which is presumably distal histidine. Dissociation constants (KD) of SC15N-/LPO, SC15N-/LPO/I-, and SC15N /LPO/CN- equilibria have been determined by 15N T1 measurements and found to be 90 +/- 5, 173 +/- 20, and 83 +/- 6 mM, respectively. On the basis of these values of KD, it is suggested that the iodide ion inhibits the binding of the thiocyanate but cyanide ion does not. The thiocyanate is shown to bind at the same site of LPO as iodide does, but the binding is considerably weaker and is away from the ferric ion. The distance of 15N of the bound thiocyanate ion from the iron is determined to be 7.2 +/- 0.2 A from the 15N T1 measurements. PMID- 2548590 TI - Nuclear Overhauser effect studies of the conformation of Co(NH3)4ATP bound to kidney Na,K-ATPase. AB - Transferred nuclear Overhauser effect measurements (in the two-dimensional mode) have been used to determine the three-dimensional conformation of an ATP analogue, Co(NH3)4ATP, at the active site of sheep kidney Na,K-ATPase. Previous studies have shown that Co(NH3)4ATP is a competitive inhibitor with respect to MnATP for the Na,K-ATPase [Klevickis, C., & Grisham, C.M. (1982) Biochemistry 21, 6979. Gantzer, M.L., et al. (1982) Biochemistry 21, 4083]. Nine unique proton proton distances on ATPase-bound Co(NH3)4ATP were determined from the initial build-up rates of the cross-peaks of the 2D-TRNOE data sets. These distances, taken together with previous 31P and 1H relaxation measurements with paramagnetic probes, are consistent with a single nucleotide conformation at the active site. The bound Co(NH3)4ATP) adopts an anti conformation, with a glycosidic torsion angle of 35 degrees, and the conformation of the ribose ring is slightly N-type (C2'-exo, C3'-endo). The delta and gamma torsional angles in this conformation are 100 degrees and 178 degrees, respectively. The nucleotide adopts a bent configuration, in which the triphosphate chain lies nearly parallel to the adenine moiety. Mn2+ bound to a single, high-affinity site on the ATPase lies above and in the plane of the adenine ring. The distances from enzyme-bound Mn2+ to N6 and N7 are too large for first coordination sphere complexes, but are appropriate for second-sphere complexes involving, for example, intervening hydrogen-bonded water molecules. The NMR data also indicate that the structure of the bound ATP analogue is independent of the conformational state of the enzyme.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548591 TI - Dissociation of double-stranded polynucleotide helical structures by eukaryotic initiation factors, as revealed by a novel assay. AB - A new technique has been applied to the study of the RNA secondary structure unwinding activity of the eukaryotic initiation factors (eIFs) 4F, 4A, and 4B. Secondary structures were generated at the 5' ends of reovirus and globin mRNA molecules by hybridization with 32P-labeled cDNA molecules 15 nucleotide residues long. The dissociation of the labeled cDNAs from the mRNAs was assayed by a gel filtration chromatography procedure which separates the free cDNAs from mRNAs and mRNA/cDNA hybrids. When the three factors were tested alone, only eIF-4F stimulated dissociation of hybrids. The combination of eIF-4A plus eIF-4B also exhibited a strong hybrid dissociating activity, which was markedly temperature dependent. Under optimum conditions, up to 90% of the hybrid structures are disrupted in 60 min. These results demonstrate for the first time that stable double-stranded regions can be melted and dissociated by eIFs. They also characterize more precisely the first step in the structure unwinding reaction. PMID- 2548592 TI - Inhibition of eukaryotic translation by nucleoside 5'-monophosphate analogues of mRNA 5'-cap: changes in N7 substituent affect analogue activity. AB - Nucleotide cap analogues of 7-methylguanosine 5'-monophosphate (m7GMP) were synthesized in which the 7-methyl moiety was replaced with 7-ethyl (e7), 7-propyl (p7), 7-isopropyl (ip7), 7-butyl (b7), 7-isobutyl (ib7), 7-cyclopentyl (cp7), 7 (carboxymethyl) (cm7), 7-benzyl (bn7), 7-(2-phenylethyl) [7-(2-PhEt)], and 7-(1 phenylethyl) [7-(1-PhEt)]. These derivatives were assayed as competitive inhibitors of capped mRNA translation in reticulocyte lysate. We observed that N7 alkyl and alicyclic substituents larger than ethyl significantly decreased the inhibitory activity of these cap analogues presumably by decreasing their affinity for cap binding proteins, which participate in the initiation of translation. This result defined a maximum size for this class of N7 substituents in the nucleotide binding domain of cap binding proteins. Like m7GMP, the N7 substituted GMP derivatives synthesized in this study were found to be predominantly in the anti conformation as determined by proton NMR analyses. However, bn7GMP and 7-(2-PhEt)GMP, which have aromatic N7 substituents, were more effective than m7GMP as competitive inhibitors of translation. The increased affinity of bn7GMP for cap binding proteins was further examined by synthesis of beta-globin mRNA containing 5'-bn7G, 5'-m7G, or 5'-e7G cap structures. These modified mRNAs were tested as translation templates. Messenger RNA capped with bn7G was observed to increase the translation activity of the template 1.8-fold relative to that of its m7G-capped mRNA counterpart. By contrast, e7G-capped mRNA was 25% less active than m7G-capped mRNA.2+V photo-cross-linking of m7G-capped mRNA to cap binding proteins PMID- 2548593 TI - Characterization of free radicals produced during oxidation of etoposide (VP-16) and its catechol and quinone derivatives. An ESR Study. AB - Spectroscopic evidence for the radical-mediated metabolism of VP-16, VP-16 catechol, and VP-16 quinone during enzymatic oxidation and autoxidation has been obtained. Autoxidation of the catechol yields the primary semiquinone together with the primary molecular product VP-16 quinone, which subsequently undergoes hydrolytic oxidation to form secondary quinones and semiquinones. Both primary and secondary phenoxyl radicals were detected during peroxidatic oxidation of VP 16. Neither the primary nor the secondary radicals react with DNA at a detectable rate. Evidence for the production of hydroxyl radical during iron-catalyzed oxidation of VP-16 catechol was obtained. These free radical reactions may have implications for the mechanism of antitumor action of VP-16. PMID- 2548594 TI - Proton nuclear magnetic resonance study of the molecular and electronic structure of the heme cavity in Aplysia cyanometmyoglobin. AB - The 1H NMR spectrum of the low-spin, cyanide-ligated ferric complex of the myoglobin from the mollusc Aplysia limacina has been investigated. All of the resolved resonances from both the hemin and the proximal histidine have been assigned by a combination of isotope labeling, spin decoupling, analysis of differential paramagnetic relaxation, and nuclear Overhauser (NOE) experiments. The pattern of the heme contact shifts is unprecedented for low-spin ferric hemoproteins in exhibiting minimal rhombic asymmetry. This low in-plane asymmetry is correlated with the X-ray-determined orientation of the proximal histidyl imidazole plane relative to the heme and provides an important test case for the interpretation of hyperfine shifts of low-spin ferric hemoproteins. The bonding of the proximal histidine is shown to be similar to that in sperm whale myoglobin and is largely unperturbed by conformational transitions down to pH approximately 4. The two observed conformational transitions appear to be linked to the titration of the two heme propionate groups, which are suggested to exist in various orientations as a function of both pH and temperature. Heme orientational disorder in the ratio 5:1 was demonstrated by both isotope labeling and NOE experiments. The exchange rate with bulk water of the proximal histidyl labile ring proton is faster in Aplysia than in sperm whale myoglobin, consistent with a greater tendency for local unfolding of the heme pocket in the former protein. A similar increased heme pocket lability in Aplysia myoglobin has been noted in the rate of heme reorientation [Bellelli, A., Foon, R., Ascoli, F., & Brunori, M. (1987) Biochem. J. 246, 787-789]. PMID- 2548595 TI - Modeling with in vitro kinetic parameters for the elaboration of transfer RNA identity in vivo. AB - A tRNA with "double identity" was created, and this tRNA was demonstrated in vitro to aminoacylate quantitatively with either of two amino acids. In contrast, acceptance of only one of these amino acids was observed in vivo, and a simple manipulation determined which one was accepted. Kinetic parameters were obtained for aminoacylation with each amino acid of the tRNA with double identity and of related tRNAs. Modeling with these parameters largely explains which amino acid specificity is observed in vivo. The results delineate some of the kinetic boundaries for the design and accommodation of tRNA sequence variations in the elaboration of identity in vivo. PMID- 2548597 TI - Carbon-13 NMR of glycogen: hydration response studied by using solids methods. AB - The carbon-13 NMR spectra of glycogen are reported by using the methods of magic angle sample spinning and high-power proton decoupling to provide a dynamic report on the glucose monomer behavior as a function of hydration. Although the glycogen behaves as a typical polymer in the dry state, addition of water makes a significant difference in the spectral appearance. Water addition decreases the carbon spin-lattice relaxation times by 2 orders of magnitude over the range from 7% to 70% water by weight. The proton-carbon dipole-dipole coupling, which broadens the carbon spectrum and permits cross-polarization spectroscopy, is lost with increasing hydration over this range. By 60% water by weight, scalar decoupling methods are sufficient to achieve a reasonably high-resolution spectrum. Further, at this concentration, the carbon spin-lattice relaxation times are near their minimum values at a resonance frequency of 50.3 MHz, making acquisition of carbon spectra relatively insensitive to intensity distortions associated with saturation effects. Though motional averaging places the spectrum in the solution phase limit, the static spectrum shows a residual broader component that would not necessarily be detected readily by using high-resolution liquid-state experiments. PMID- 2548596 TI - Phosphatidylinositol 4,5-bisphosphate competitively inhibits phorbol ester binding to protein kinase C. AB - Calcium phospholipid dependent protein kinase C (PKC) is activated by diacylglycerol (DG) and by phorbol esters and is recognized to be the phorbol ester receptor of cells; DG displaces phorbol ester competitively from PKC. A phospholipid, phosphatidylinositol 4,5-bisphosphate (PIP2), can also activate PKC in the presence of phosphatidylserine (PS) and Ca2+ with a KPIP2 of 0.04 mol %. Preliminary experiments have suggested a common binding site for PIP2 and DG on PKC. Here, we investigate the effect of PIP2 on phorbol ester binding to PKC in a mixed micellar assay. In the presence of 20 mol % PS, PIP2 inhibited specific binding of [3H]phorbol 12,13-dibutyrate (PDBu) in a dose-dependent fashion up to 85% at 1 mol %. Inhibition of binding was more pronounced with PIP2 than with DG. Scatchard analysis indicated that the decrease in binding of PDBu in the presence of PIP2 is the result of an altered affinity for the phorbol ester rather than of a change in maximal binding. The plot of apparent dissociation constants (Kd') against PIP2 concentration was linear over a range of 0.01-1 mol % with a Ki of 0.043 mol % and confirmed the competitive nature of inhibition between PDBu and PIP2. Competition between PIP2 and phorbol ester could be demonstrated in a liposomal assay system also. These results indicate that PIP2, DG, and phorbol ester all compete for the same activator-receiving region on the regulatory moiety of protein kinase C, and they lend support to the suggestion that PIP2 is a primary activator of the enzyme. PMID- 2548598 TI - Conformational changes of plasma fibronectin detected upon adsorption to solid substrates: a spin-label study. AB - Changes in local environment of the free sulfhydryl groups in plasma fibronectin upon adsorption of the protein to polystyrene beads have been examined by electron spin resonance (ESR) spin-label spectroscopy. The two free sulfhydryl groups per subunit of plasma fibronectin were modified chemically with an [15N, 2H]maleimide spin-label. For soluble fibronectin, both free sulfhydryl groups are shown to be in confined environments as evidenced from the labeled protein exhibiting a strongly immobilized ESR spectrum as described previously using [14N, 1H]maleimide spin-labels [Lai, C.-S., & Tooney, N. M. (1984) Arch. Biochem. Biophys. 228, 465-473]. When the labeled protein was adsorbed to the beads, half of the strongly immobilized component was found to convert into a weakly immobilized component, a result indicating that one of the two labeled sites becomes exposed and exhibit a fast tumbling motion. Experiments conducted using various spin-labeled fibronectin fragments suggest that the newly exposed labeled site is located between the DNA-binding and the cell-binding regions of the molecule. The data obtained indicate that, upon adsorption to polystyrene beads, plasma fibronectin undergoes a conformational change through which the buried free sulfhydryl group near the cell-binding region of the molecule is exposed. This observation may have important implications regarding the expression of cell adhesive properties of the fibronectin molecule. PMID- 2548599 TI - Ionic strength dependence of cytochrome c structure and Trp-59 H/D exchange from ultraviolet resonance Raman spectroscopy. AB - Ultraviolet resonance Raman spectra are reported for cytochrome c (cyt c) in FeII and FeIII oxidation states at low (0.005 M) and high (0.9-1.5 M) ionic strength. With 200-nm excitation the amide band intensities are shown to remain constant, establishing that redox state and ionic strength have no influence on the alpha helical content. The tyrosine 830/850-cm-1 doublet, however, shows a loss in 830 cm-1 intensity at I = 0.005 M for the FeIII protein, suggesting a weakening or a loss of H-bonding from an internal tyrosine, probably Tyr-48, which is H-bonded to a heme propionate group in cyt c crystals. Excitation profiles of tryptophan peak at approximately 229 nm for both FeII and FeIII forms of cyt c, but at approximately 218 nm for aqueous tryptophan. The approximately 2200-cm-1 red shift of the resonant electronic transition is attributed to the Trp-59 residue being buried and H-bonded. Consistent with this Trp environment, the H-bond sensitive 877-cm-1 Trp band is strong and sharp, and the 1357/1341-cm-1 doublet has a large intensity ratio, approximately 1.5, for both FeII and FeIII cyt c. The 877-cm-1-band frequency shifts to 860 cm-1 when the Trp indole proton is replaced by a deuteron. This band was used to show that Trp H/D exchange in D2O is much faster for FeIII than FeII cyt c. The half-time for exchange at room temperature is estimated to be approximately 30 and approximately 5 h, respectively, for FeII and FeIII when examined at I = 0.005.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548600 TI - Effects of temperature and glycerol on the resonance Raman spectra of cytochrome c peroxidase and selected mutants. AB - The high-frequency resonance Raman spectra of FeIII yeast native cytochrome c peroxidase (CCP) and five of its mutants [CCP(MI), Phe-51, Leu-48, Lys-48, Asn 235, and Phe-191] were recorded in phosphate buffer, pH 7.0, and in glycerol/phosphate mixtures at 295 and 10 K. Glycerol induces heme coordination changes in some of the CCP mutants at room temperature. It apparently weakens the binding of the Fe atom to ligands in the distal heme cavity and drives the heme toward the 5-coordinate, high-spin state. At 10 K, native CCP and all the mutants (except Phe-51 which remains 6-coordinate, high-spin) show various distributions of spin and coordination states which differ from those observed at 295 K. Upon cooling in phosphate buffer, pH 7, and to a much lesser extent in 66% glycerol/phosphate, an internal strong-field ligand is coordinated to the Fe. A likely candidate is H2O-595, which could become a strong-field ligand on H bonding and/or proton transfer to H2O-648, and/or the distal His-52. However, distal His-52 itself cannot be ruled out as the coordinating ligand considering that the Phe-51 mutant, which binds H2O-595 at room temperature, does not show a large 6-coordinate, low-spin component at 10 K like the other mutants. These results clearly indicate that the Fe coordination in CCP and its mutants is sensitive to both temperature and solvent composition. PMID- 2548601 TI - Structure of the active site of sulfite oxidase. X-ray absorption spectroscopy of the Mo(IV), Mo(V), and Mo(VI) oxidation states. AB - The active site of sulfite oxidase has been investigated by X-ray absorption spectroscopy at the molybdenum K-edge at 4 K. We have investigated all three accessible molybdenum oxidation states, Mo(IV), Mo(V), and Mo(VI), allowing comparison with the Mo(V) electron paramagnetic resonance data for the first time. Quantitative analysis of the extended X-ray absorption fine structure indicates that the Mo(VI) oxidation state possesses two terminal oxo (Mo = O) and approximately three thiolate-like (Mo-S-) ligands and is unaffected by changes in pH and chloride concentration. The Mo(IV) and Mo(V) oxidation states, however, each have a single oxo ligand plus one Mo-O- (or Mo-N less than) bond, most probably Mo--OH, and two to three thiolate-like ligands. Both reduced forms appear to gain a single chloride ligand under conditions of low pH and high chloride concentration. PMID- 2548602 TI - Formation and utilization of formyl phosphate by N10-formyltetrahydrofolate synthetase: evidence for formyl phosphate as an intermediate in the reaction. AB - N10-Formyltetrahydrofolate synthetase from bacteria and yeast catalyzes a slow formate-dependent ADP formation in the absence of H4folate. The synthesis of formyl phosphate by the enzyme was detected by trapping the intermediate as formyl hydroxamate. That the "formate kinase" activity was part of the catalytic center of N10-formyltetrahydrofolate synthetase was shown by demonstrating coordinate inactivation of the "kinase" and synthetase activities by heat and a sulfhydryl reagent, similar effects of monovalent cations, similar Km values for substrates, and similar Ki values for the inhibitor phosphonoacetaldehyde for both activities. The relative rates of the kinase activities for the bacterial and yeast enzymes are about 10(-4) and 4 x 10(-6) of their respective synthetase activities. These slow rates for the kinase reaction can be explained by the slow dissociation of ADP and formyl phosphate from the enzyme. This conclusion is supported by rapid-quench studies where a "burst" of ADP formation (6.4 s-1) was observed that is considerably faster than the steady-state rate (0.024 s-1). The demonstration of enzyme-bound products by a micropartition assay and the lack of a significant formate-stimulated exchange between ADP and ATP provide further evidence for the slow release of the products from the enzyme. The synthesis of N10-CHO-H4folate when H4folate was added to the E-formyl phosphate-ADP complex is also characterized by a "burst" of product formation. The rate of this burst phase at 5 degrees C occurs with a rate constant of 18 s-1 compared to 14 s-1 for the overall reaction at the same temperature. These results provide further evidence for formyl phosphate as an intermediate in the reaction and are consistent with the sequential mechanism of the normal catalytic pathway. Positional isotope exchange experiments using [beta,gamma-18O]ATP showed no evidence for exchange during turnover experiments in the presence of either H4folate or the competitive inhibitor pteroyltriglutamate. The absence of scrambling of the 18O label as observed by 31P NMR suggests that the central complex may impose restraints to limit free rotation of the P beta oxygens of the product ADP. PMID- 2548603 TI - Human and rat malignant-tumor-associated mRNAs encode stromelysin-like metalloproteinases. AB - Rat transin and human stromelysin 2 mRNAs, which have been associated with malignant tumors, code for potential proteins with significant sequence homology to the metalloproteinases collagenase and stromelysin. We have used an expression system that allows easy purification of these proteins after transfection of COS cells with a vector containing the corresponding cDNA. This system has allowed us to prepare transin and stromelysin 2 as active proteinases that are inhibited by inhibitors of metalloproteinases. Further analysis of these enzymes indicates that they degrade several components of the extracellular matrix including collagen types III, IV, and V and fibronectin, as well as gelatins formed from several denatured collagen types. In addition, both transin and stromelysin 2 are capable of activating procollagenase in vitro. Thus, in malignant tumors these proteinases may act, both directly and indirectly, to degrade the extracellular matrix and permit tumor invasion of neighboring tissues. PMID- 2548604 TI - Molecular organization of developmentally regulated Dictyostelium discoideum ubiquitin cDNAs. AB - Dictyostelium discoideum ubiquitin mRNAs are regulated in a complex fashion during spore germination and multicellular development. Species of mRNA of 1900, 1400, 1100, 840, 580, and 500 nucleotides (nt) are found which are expressed differentially during different stages of development. DNA blot analysis indicates that ubiquitin genes constitute a multigene family of at least six genes. cDNAs representing all the ubiquitin mRNA transcripts were isolated and sequenced. The Dictyostelium mRNAs are organized as tandem repeats of the 76 amino acid ubiquitin unit (228 nt). We isolated one cDNA containing seven of these tandem repeats, and two different five- and three-repeat cDNAs. In addition, 2 cDNAs containing a single ubiquitin repeat fused at its 3' end to an unrelated 52 and 78 amino acid extension were identified. There is a remarkable similarity in the sequences of the non-ubiquitin extensions among yeast and mammalian counterparts. The extensions are very basic, containing approximately 30% lysine/arginine. Another common feature of these proteins is the presence of a common structural motif containing cysteine residues at conserved positions, suggesting a metal binding domain that matches a consensus sequence of Xenopus transcription factor TFIIIA and other nucleic acid binding proteins. The characterization of ubiquitin cDNAs and genomic sequences in D. discoideum now makes the understanding of its developmental regulation feasible. PMID- 2548605 TI - Binding of actinomycin D to [d(ATCGAT)]2: NMR evidence of multiple complexes. AB - Actinomycin D (actD) binds to the oligonucleotide [d(ATCGAT)]2 with a hypochromatic and red-shifted visible absorbance band compared to free drug and a CD spectrum with double negative bands at 460 and 385 nm. These spectral features are similar to those of the actD-[d(ATGCAT)]2 complex, while actD-[d(AT)5]2 gives spectra similar to those of free drug. Upon dilution or raising the temperature, the spectral characteristics accompanying complex formation disappear in the actD [(ATCGAT)]2 sample but remain in the actD-[d(ATGCAT)]2 complex under the same experimental conditions. These results suggest that (a) sequence-specific binding of actD occurs with [d(ATCGAT)]2 but not with [d(AT)5]2, (b) the binding is not as strong as with [d(ATGCAT)]2, and (c) actD binds [d(ATCGAT)]2 with the same mechanism as it binds [d(ATGCAT)]2, i.e., by intercalation. From NMR spectra of the actD-[d(ATCGAT)]2 complex, three types of signals can be detected below 20 degrees C, one major and two minor ones. At higher temperatures, exchange between the two minor ones becomes fast enough that only one type of minor signal was seen. Partial resonance assignments were made by using 2D nuclear Overhauser effect (NOE) and 2D homonuclear Hartmann-Hahn (HOHAHA) experiments. Proton chemical shift changes of the major complex are consistent with actD chromophore ring intercalation between hexamer base pairs. Data from NOE-detected dipolar interactions between actD and [d(ATCGAT)]2 protons were interpreted in terms of a major complex with the actD chromophore ring system intercalated at the CG position and minor complexes with the drug intercalated off center at the GA positions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548606 TI - Quantification of DNA structure from NMR data: conformation of d-ACATCGATGT. AB - Resonance assignments of nonexchangeable base and sugar protons have been obtained in double-helical d-ACATCGATGT by using two-dimensional correlated spectroscopy (COSY) and nuclear Overhauser enhancement spectroscopy (NOESY). The exchangeable imino protons have been assigned on the basis of their chemical shifts. The characteristic phase-sensitive multiplet patterns of the intrasugar cross-peaks in the omega 1-scaled COSY spectrum have been used to estimate several scalar coupling constants (J). The information on the J values combined with the intranucleotide COSY cross-peak intensities has been used to identify sugar puckers of individual nucleotide units. In most cases, the deoxyribofuranose rings are found to adopt a conformation close to O4'-endo. Spin diffusion has been monitored from the buildup of the normalized volumes of NOE cross-peaks in NOESY spectra as a function of mixing time. A set of 52 intranucleotide and internucleotide proton-proton distances have been estimated by using low mixing time NOESY spectra (tau m = 40 and 80 ms). The estimated intrasugar proton-proton distances rule out possibilities of existence of a fast equilibrium between C2'-endo and C3'-endo conformations. Intranucleotide proton proton distances combined with the knowledge of sugar puckers have been used to fix the glycosidic bond torsion angle (chi). For this purpose, simulated distance contours depicting the dependence of intranucleotide proton-proton distances on pseudorotational phase angle (P) and glycosidic bond torsion angle (chi) have been used. Further, the proton homonuclear (J, delta) spectrum has been used to monitor the 31P-1H heteronuclear couplings, which are preserved in the omega 2 projection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548607 TI - Persistence of the alpha-helix stop signal in the S-peptide in trifluoroethanol solutions. AB - alpha-Helix formation in the S-peptide (residues 1-19 of ribonuclease A) was studied in detail by use of two-dimensional 1H nuclear magnetic resonance to monitor the effects of 2,2,2-trifluoroethanol (TFE) at 0 degrees C and pH* 2.07. TFE stabilizes the S-peptide alpha-helix. Helix formation by a particular amino acid was monitored by the chemical shifts of the C alpha, C beta, and C gamma protons while increasing the concentration of TFE: large changes in chemical shift of a particular residue indicate that it is induced to go helical, whereas small chemical shift changes indicate little helix formation. Residues Thr-3 to Met-13 undergo chemical shift changes consistent with helix formation, whereas the other residues do not. Earlier work [Kim, P. S., & Baldwin, R. L. (1984) Nature 307, 329-334] reported that residues Thr-3 to His-12 become helical in aqueous solution. The existence of a "helix stop signal" was inferred from this behavior. We thus conclude that this helix stop signal persists in TFE solutions. PMID- 2548608 TI - End-label fingerprintings show that the N- and C-termini of actin are in the contact site with gelsolin. AB - Gelsolin was cleaved by chymotrypsin or thermolysin into an N-terminal Mr 45,000 fragment (45N) and a C-terminal Mr 38,000 fragment (38C). The N-terminal half was further cleaved into two fragments with Mr 17,000 (17N) and Mr 28,000 (28N). These fragments were complexed with actin and cross-linked with 1-ethyl-3-[3 (dimethylamino)prophyl]carbodiimide (EDC) to introduce covalent bonds into their contact sites. The location of these bonds was mapped along the actin sequence by end-label fingerprinting with highly sensitive probes for the N- and C-termini of actin. The mapping studies revealed that two gelsolin N-terminal fragments (17N and 28N) were cross-linked with the actin C-terminal segment. The result indicates that the actin N- and C-terminal segments are in the binding site of gelsolin. PMID- 2548609 TI - Spin equilibria in human methemoglobin: effects of bezafibrate and inositol hexaphosphate as measured by susceptometry and visible spectroscopy. AB - The effects of inositol hexaphosphate (IHP) and a second allosteric effector, bezafibrate, on the spin-state equilibria of the mixed-spin derivatives of ferric human hemoglobin A are examined. Changes in spin-state equilibrium are monitored by measuring absorption spectra in the visible region (460-700 nm) as well as by direct measurements of magnetic susceptibility by means of a superconducting fluxmeter. The addition of IHP at pH 6.5 results in a measurable shift in the spin equilibria of these derivatives toward higher spin. However, the addition of bezafibrate in the presence of IHP results in still larger shifts toward the high spin form. The changes in the free energies of the spin-state equilibria resulting from the combination of these two effectors are similar in magnitude to that which results from the R-state to T-state transition in carp hemoglobin. PMID- 2548610 TI - Spin equilibrium in human methemoglobin: effects of inositol hexaphosphate and bezafibrate as measured by resonance Raman spectroscopy. AB - The spin equilibria of several derivatives of human methemoglobin were probed by resonance Raman scattering. The intensity of lines in the Raman spectrum gives a measure of the high-spin (S = 5/2) to low-spin (S = 1/2) ratio which agrees well with the spin equilibria determined from direct magnetic susceptibility measurements. The addition of bezafibrate (BZF) to methemoglobin in the absence of organic phosphate, IHP, has very little effect on the spin equilibrium, whereas in the presence of IHP it augments the change in spin significantly. When both IHP and BZF are added to the mixed-spin derivatives (H2O, SCN-, OCN-, and NO2-) of human methemoglobin, the spin equilibrium is shifted toward higher spin by about 700 cal/mol, similar to the spin change detected in derivatives of carp methemoglobin upon addition of IHP alone. These data support a general mechanism for the allosteric transition in which a constant fraction of the cooperative energy (approximately 20%) is detected at the heme of the ferric ligand-bound forms. PMID- 2548611 TI - Involvement of calcium and protein kinase C in the activation of the Na+/H+ exchanger in cultured bovine aortic endothelial cells stimulated by extracellular ATP. AB - We have studied the activation of the Na+/H+ exchanger which leads to the intracellular alkalinization in cultured bovine aortic endothelial cells stimulated by extracellular ATP. The alkalinization induced by ATP was largely dependent on extracellular Ca2+ and the rate of alkalinization was decreased by about 60% in the absence of extracellular Ca2+. ATP caused a rapid and transient increase and a subsequent sustained increase of the intracellular Ca2+ concentration ([Ca2+]i) in the Ca2+ buffer, while only the rapid and transient increase of [Ca2+]i was observed in the absence of extracellular Ca2+. The Ca2+ depleted cells prepared by incubation in Ca2+-free buffer containing 0.1 mM EGTA showed only a slight increase of [Ca2+]i with no alkalinization on stimulation by ATP. The alkalinization was inhibited by 1-(5-isoquinolinesulfonyl)-2 methylpiperazine dihydrochloride (H-7), an inhibitor of protein kinase C, but not by another isoquinoline analogue (HA 1004), which has a less inhibitory effect on the kinase. Phorbol 12-myristate 13-acetate also induced the alkalinization by the activation of the Na+/H+ exchanger. Neither dibutyryl cyclic AMP nor dibutyryl cyclic GMP affected the alkalinization induced by ATP. Treatment of the cells by pertussis and cholera toxins had no effect on the alkalinization. The results suggest that the increase in [Ca2+]i is essential for the ATP-induced activation of the Na+/H+ exchanger in cultured bovine aortic endothelial cells and a protein kinase C-dependent pathway is involved in the activation. PMID- 2548612 TI - Parathyroid hormone inhibition of Na+/phosphate cotransport in OK cells: requirement of protein kinase C-dependent pathway. AB - Parathyroid hormone (PTH) inhibits sodium/phosphate (Na+/Pi) cotransport across the apical membrane of opossum kidney (OK) cells principally through two pathways. First, cAMP stimulation and activation of protein kinase A; second, diacylglycerol release and stimulation of protein kinase C. Studies were designed to determine the importance of these regulatory cascades. Down-regulation of protein kinase C with prolonged phorbol ester (12-O-tetradecanoylphorbol 13 acetate (TPA] treatment leads to a refractory state in which the cells do not respond to PTH (10(-8) M), cAMP (10(-4) M) or rechallenge of TPA (200 nM) even though Na+/Pi cotransport is similar to control cells (8.1 +/- 0.1 nmol.mg-1 protein.5 min-1). Staurosporine, an inhibitor of protein kinase C, resulted in the complete inhibition of PTH, cAMP and TPA action in a dose-dependent manner. PTH, cAMP and TPA were additive below maximal concentrations, but had no further effect at maximal agonist concentrations. These results suggest that protein kinase C activity is important in PTH-mediated inhibition of Na+/phosphate cotransport in OK cells. PMID- 2548613 TI - Parathyroid hormone inhibition of Na+/phosphate cotransport in OK cells: intracellular [Ca2+] as a second messenger. AB - Parathyroid hormone increases cellular cAMP, 1,2-diacylglycerol, inositol 1,4,5 trisphosphate and cytosolic Ca2+ concentration ([Ca2+]i) in OK cells. In the present study, we determined the importance of the PTH-dependent increase in [Ca2+]i in the control of sodium-dependent phosphate (Na+/Pi) cotransport. PTH (10(-7) M) results in a transient increase in [Ca2+]i from basal levels of 67 +/- 4 nM to maximal concentrations of 190 +/- 9 nM. The increase in [Ca2+]i was dose dependent with half-maximal increases at about 5.10(-8) M PTH. These hormone levels were 10(3)-fold higher than that required for half-maximal inhibition of Na+/Pi cotransport. Clamping [Ca2+]i with either intracellular Ca2+ chelators or by ionomycin in the presence of high concentrations of extracellular Ca2+ did not alter PTH-dependent inhibition of Na/Pi cotransport. Nor did indomethacin, an inhibitor of the cyclooxygenase pathway, influence the hormonal inhibition of cotransport. Accordingly, these data suggest that changes in [Ca2+]i and/or activation of the phospholipase A2 and the cyclooxygenase pathways are not involved in signal induction of the PTH-mediated control of Na+/Pi cotransport. PMID- 2548615 TI - The effect of acetylcholine on inositol lipid metabolism and intracellular calcium concentrations in bovine anterior pituitary cells. AB - Acetylcholine (ACh) increased the intracellular calcium concentration in bovine anterior pituitary cells. In the presence of the calcium channel antagonists verapamil (20 microM) or nitrendepine (1 microM) the increase in calcium was partially inhibited but showed both transient and sustained components. In the presence of EGTA (2.5 mM) only the transient component was observed. ACh also decreased inositol radioactivity in phosphatidylinositides and increased it in inositol phosphates. It is concluded that the increase in calcium caused by acetylcholine requires both the entry of external calcium and mobilisation of internal calcium. Replacement of external sodium by N-methyl-D-glucamine inhibited the rises in calcium and inositol phosphate labelling in response to ACh. Tetrodotoxin (3 microM) or ouabain (50 microM) did not affect either response to ACh. Verapamil did not affect the calcium rise induced by ACh in the absence of external sodium. The phorbol ester PMA (10 nM) caused a transient rise in calcium and inhibited the calcium rise caused by acetylcholine: it did not modify the effect of acetylcholine on inositol phosphates. The dependence of the stimulation of external calcium entry and inositol phosphate production on external sodium ions and protein kinase C is discussed. PMID- 2548614 TI - The effects of p21N-ras expression in NIH-3T3 cells upon cyclic AMP metabolism. AB - The effects of overexpression of p21N-ras upon cyclic AMP metabolism have been examined in the inducible T15 cell line. In cells overexpressing the N-ras gene product, beta-adrenergic stimulation of cyclic AMP generation was reduced. The reduction was more pronounced the longer the ras gene was expressed and in chronically transformed cells a reduction in forskolin-stimulated cyclic AMP generation was also observed. The transformed cells exhibited a reduction in beta adrenergic binding sites, but no change in the apparent EC50 for agonist induced cyclic AMP generation. Treatment of the cells with dibutyryl cyclic AMP induced a dose-dependent inhibition of proliferation, with the transformed cells being more sensitive than the control cells. PMID- 2548616 TI - Inhibitory action of cyclic AMP on inositol 1,4,5-trisphosphate-induced Ca2+ release in saponin-permeabilized platelets. AB - Ca2+ release triggered by inositol 1,4,5-trisphosphate (IP3) has been measured in saponin-permeabilized human platelets with quin2 or 45Ca2+. Ca2+ was sequestered by intracellular organelles in the presence of ATP, and IP3 released half of the sequestered Ca2+. The addition of cyclic AMP (cAMP) to permeabilized platelets transiently accelerated Ca2+ sequestration, but did not alter the steady-state level. In contrast, IP3-induced Ca2+ release was greatly inhibited by cAMP. Phorbol myristate acetate, an activator of protein kinase C did not affect IP3 induced Ca2+ release. These results indicate that cAMP may be involved in the regulation of IP3-induced Ca2+ release in human platelets. PMID- 2548617 TI - Further study on the role of Mg2+ in lipid-protein interaction in reconstituted porcine heart mitochondrial H+-ATPase. AB - Porcine heart mitochondrial H+-ATPase was reconstituted by cholate dialysis method in liposomes containing neutral (PC, PE), acidic (PG, PI, PA, PS, DPG) or neutral and acidic phospholipids. The Mg2+ effect on the ATPase activity and its sensitivity to oligomycin, ATP-induced delta psi and delta pH formation was observed for the proteoliposomes containing acidic but not neutral phospholipids. Maleimide spin labels with varying arm lengths or bromoacetamide spin probe were used to monitor the conformational difference of H+-ATPase in the Mg2+-containing and Mg2+-'free' samples. A difference in W/S ratio (weakly immobilized/strongly immobilized component in the ESR spectra) could be detected for the F0.F1 containing and F1-depleted, (F0)-containing proteoliposomes, suggesting conformational difference in the F0-F1 complex and F0 portion induced by the Mg2+ effect. A conformational change of the beta-subunits in the F1 portion was also deduced from the ATP-induced fluorescence quenching of aurovertin-complex for Mg2+-containing samples. The results obtained are in favor of our previous assumption that Mg2+ may play its role by altering the physical state of the lipid bilayer, which would induce a conformational change in F0 (buried in the lipid core), which in turn is transmitted to the catalytic F1, resulting in a higher enzyme activity. PMID- 2548619 TI - Physical biochemistry of a liposomal amphotericin B mixture used for patient treatment. AB - There seems little doubt now that intravenous liposomal amphotericin B can be a useful treatment modality for the management of immunocompromised patients with suspected or proven disseminated fungal infections. Interestingly, the very significant reduction in toxicity reported when amphotericin B is part of a bilayer membrane is closely tied to the physical characteristics of the liposomes involved, although these are poorly understood at the molecular level. We record here an examination by spectroscopy and freeze-etch electron microscopy of unsonicated amphotericin B multilamellar vesicles prepared along the lines that we and others have followed for samples used in clinical trials and preclinical in vivo or in vitro studies. Our study has focussed on liposomes of 7:3 dimyristoylphosphatidylcholine/dimyristoylphosphatidylglycerol (DMPC/DMPG) bearing 0-25 mol% amphotericin B, since this lipid mixture has been the choice for the first clinical trials. Phase transition behaviour of these liposomes was examined by electron paramagnetic resonance (EPR) spectroscopy of a nitroxide spin label partitioning into the bilayers. The same experiments were then performed on similarly prepared liposomes of the disaturated species, dipalmitoylphosphatidylcholine (DPPC), and the diunsaturated species, dielaidoylphosphatidylcholine (DEPC). Partial phase diagrams were constructed for each of the lipid/drug mixtures. Melting curves and derived phase diagrams showed evidence that amphotericin B is relatively immiscible with the solid phase of bilayer membranes. The phase diagram for DEPC/amphotericin B was very similar to that of DPPC/amphotericin B, and both exhibited less extensive temperature ranges of phase separation than did the 7:3 DMPC/DMPG mixture with amphotericin B. Between 25 and 37 degrees C the measured fluidity of the 7:3 DMPC/DMPG liposomes was similar to that of the (unsaturated fatty acid) DEPC liposomes, and considerably higher than that seen for (saturated fatty acid) DPPC liposomes. Preparations of 7:3 DMPC/DMPG, DPPC, and DEPC containing 0-25 mol% amphotericin B were examined by freeze-etch electron microscopy at 35 and 22 degrees C (to cover the temperature range of the mammalian body core and periphery). The same liposome features were present in all three liposome types studied. The appearance of individual liposomes at x 100,000 magnification reflected their molecular characteristics, which were found to be significantly heterogeneous within each batch. The lipid/drug structures were bilayer in nature, although liposomes showing considerable disruption were common, particularly at the highest drug concentrations.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2548618 TI - The Rhodospirillum rubrum cytochrome bc1 complex: peptide composition, prosthetic group content and quinone binding. AB - A cytochrome bc1 complex, essentially free of bacteriochlorophyll, has been purified from the photosynthetic purple non-sulfur bacterium Rhodospirillum rubrum. The complex catalyzes electron flow from quinol to cytochrome c (turnover number = 75 s-1) that is inhibited by low concentrations of antimycin A and myxothiazol. The complex contains only three peptide subunits: cytochrome b (Mr = 35,000); cytochrome c1 (Mr = 31,000) and the Rieske iron-sulfur protein (Mr = 22,400). Em values (pH 7.4) were measured for cytochrome c1 (+320 mV) and the two hemes of cytochrome b (-33 and -90 mV). Electron flow from quinol to cytochrome c is inhibited when the complex is pre-illuminated in the presence of a ubiquinone photoaffinity analog (azido-Q). During illumination, the azido-Q becomes covalently attached to the cytochrome b peptide and, to a lesser extent, to cytochrome c1. PMID- 2548621 TI - Composition and structure of lipopolysaccharide-human plasma low density lipoprotein complex. Analytical ultracentrifugation, 31P-NMR, ESR and fluorescence spectroscopy studies. AB - Complexes of Salmonella typhimurium lipopolysaccharide toxin (LPS) with low density lipoproteins (LDL) prepared in vitro have been analyzed. LPS-LDL complexes were found to comprise approx. 0.24 mg LPS/mg LDL protein. The major protein of complexes was apolipoprotein apoB-100 (greater than or equal to 90 95%). Incorporation of LPS molecules into LDL was accompanied by small changes in lipid composition, i.e. the phosphatidylcholine content was diminished by approx. 11% and the free fatty acid concentration was raised 2-fold. Analytical ultracentrifugation showed that insertion of LPS into LDL results in the increase of a portion of particles with higher density (lower flotation coefficient) compared to initial LDL. As was evidenced by ESR, in LPS-LDL complexes, the phospholipid hydrocarbon chains are more ordered than in LDL. 31P-NMR spectra indicated that in LPS-LDL complexes the mobility of phospholipid polar headgroups is restricted in comparison with LDL. Application of the shift reagent (Pr3+) revealed that phospholipid molecules form a monolayer structure on the surface of complexes. Upon binding of LPS to LDL, a maximum of the apoB intrinsic fluorescence was slightly red-shifted (1-2 nm) which may testify that the localization of apoB remains nearly unchanged. For LPS-LDL complexes, the accessibility of apoB fluorophores to quenchers (I-, Cs+, acrylamide) did not dramatically differ from that of LDL. It is concluded that rather large amounts of LPS (about 9-10 molecules) can accommodate in one LDL particle without severely perturbing its original composition and structure. Moreover, in the LPS LDL complexes, oligosaccharide chains of LPS screen notably neither phospholipid polar headgroups nor, what is very important, apoB. LPS-LDL complexes are suggested to be able in vivo to bind to cellular apoB/E receptors, possible LPS receptors and scavenger-receptors of macrophages (monocytes). PMID- 2548620 TI - Inhibition of cyclic AMP phosphodiesterase activity of human blood platelet membrane by ADP. AB - Purified human blood platelet membrane showed the presence of one low Km (1.1 microM) and one high Km (5.0 microM) cyclic AMP phosphodiesterase(s). Incubation of platelet-rich plasma or gel-filtered platelets with ADP (4.0 microM), a well known platelet aggregating agent, resulted in the inhibition of phosphodiesterase activity of the isolated membrane by 25% in 5 min at 23 degrees C. A Lineweaver Burk plot of the enzymic activity of the membrane preparation showed that ADP specifically inhibited the low Km (1.1 microM) phosphodiesterase by reducing the Vmax from 241 to 176 pmol/mg per min with concomitant lowering of Km to 0.5 microM. In contrast, neither the high Km (5.0 microM) enzymic activity of the membrane preparation nor the phosphodiesterase activities of the cytosolic fraction of the ADP-treated platelets was affected. This effect of ADP, which was independent of platelet aggregation, reached maximal level within 5 min of incubation. When platelet-rich plasma was incubated longer in the presence of nucleotide, the inhibition of phosphodiesterase activity began to decrease, and after 20 min of incubation approx. 90% of the original enzymic activity was regained. The incubation of platelet-rich plasma with 4.0 microM ADP also increased the cyclic AMP level to twice the basal level. The effect of ADP on the phosphodiesterase activity could be demonstrated only by incubating the intact platelets with the nucleotide. The treatment of isolated membrane from platelets, previously unexposed to ADP, with the nucleotide did not inhibit the enzymic activity. The inhibition of phosphodiesterase by the nucleotide in the absence of stirring, as expected, resulted in the inhibition of platelet aggregation when these cells were subsequently stirred with 1-epinephrine or an increased concentration of ADP. PMID- 2548623 TI - Human prostasome membranes exhibit very high cholesterol/phospholipid ratios yielding high molecular ordering. AB - Lipid analysis and ESR studies were carried out on prostasomes isolated from human semen. Cholesterol plus phospholipids amounted to approximately 0.80 mumol per mg protein with a striking quantitative domination of cholesterol over the phospholipids, the molar ratios of cholesterol/sphingomyelin/glycerophospholipids being 4:1:1. Saturated and monounsaturated fatty acids were dominating both in the glycerophospholipids and in sphingomyelin. The order parameters, S, deduced from ESR spectra of spin-labelled fatty acids incorporated into prostasome membranes order parameters, S, deduced from ESR spectra of spin-labelled fatty acids incorporated into prostasome membranes were very high, viz. 0.75 for 5 doxylstearic acid and 0.30 for 16-doxylstearic acid at 25 degrees C. Slightly lower values were obtained for the spin-labelled fatty acids when they were incorporated into dispersions of extracted prostasome lipids or into synthetic lipid mixtures of similar composition. The highly ordered lipids in the prostasome membrane thus seemed to be minimally perturbed by proteins in the membrane and ESR spectra showed no signs of immobilized lipids. PMID- 2548622 TI - Thrombin-induced activation of calcium transport pathways and their role in platelet functions. AB - In human platelets thrombin-induced calcium release from intracellular stores, the consequent influx of extracellular calcium, as well as their role in the aggregation and ATP-secretion reactions were examined. In indo-1-loaded platelets intracellular calcium release was studied in the presence of excess EGTA in the incubation medium, while calcium influx was followed after a rapid repletion of external calcium. After thrombin-stimulation both calcium release and calcium influx produced about the same peak levels of cytoplasmic free calcium but in the first case it was only a transient response, while in the latter one a sustained calcium signal was observed. Increased calcium influx could be evoked for several minutes after the addition of thrombin, it was selectively inhibited by Mg2+ (20 mM) and Ni2+ (1 mM) ions, by neomycin and by PCMB, a non-penetrating SH-group reagent. This calcium influx was practically insensitive to organic calcium channel blockers. Thrombin-induced platelet aggregation was only partial in the absence of external calcium, even if excess magnesium was present in the media, while the aggregation response became complete if external calcium was repleted. A significantly reduced aggregation could be seen in calcium-containing media if calcium influx was selectively inhibited. Platelet ATP-secretion under the same conditions did not depend on external calcium or on calcium influx. These data indicate that in thrombin-stimulated platelets the opening of specific plasma membrane calcium channels can be selectively modulated and these channels play a major role in the development of a full-scale aggregation. PMID- 2548624 TI - Stabilization of detergent-solubilized Ca2+-ATPase by poly(ethylene glycol). AB - The (Ca2+ + Mg2+)-ATPase from sarcoplasmic reticulum (SR) has been solubilized with 1-alkanoyl propanediol-3-phosphorylcholines with chainlengths ranging between 8 and 12 C atoms. A marked dependence of the ATPase activity upon the chainlength was found, indicating that alkyl chainlengths with 12 C atoms are necessary for retention of activity. Addition of poly(ethylene glycol) to the eluting buffers used for gel filtration of the ATPase-detergent micelles was found to increase the activity and the long-term stability significantly. In the presence of Ca2+, the elution volume indicated an ATPase dimer, whereas in the absence of Ca2+ the elution volume indicated a monomeric solution. The purity of the preparations after gel filtration was improved by subsequent chromatography with a hydroxyapatite column. PMID- 2548625 TI - The occupancy of glycoprotein IIb-IIIa complex modulates thrombin activation of human platelets. AB - Platelet membrane glycoprotein (GP IIb-IIIa), besides its activity as adhesive protein receptor, displays a number of properties supporting its involvement in the mechanisms of transduction of the activation signal. Recently we have observed that GP IIb-IIIa ligands, mostly fibrinogen, inhibit Ca2+ movement and cytoskeleton reorganization caused by mild platelet activation. These findings led us to investigate the effect of GP IIb-IIIa ligands on agonist-induced platelet responses, with particular attention to the two major messenger generating systems, involving the activation of phospholipase C and the inhibition of cAMP production. In this paper we demonstrate that the occupancy of the major adhesive protein receptor on the platelet surface modulates the phosphatidylinositol cycle decreasing the amount of IP3, IP2 and IP produced after mild platelet activation as well as the pattern of protein phosphorylation. The platelet cAMP content of activated platelets was also affected and kept higher when evaluated under the same experimental conditions. Our data provide evidence for a role of fibrinogen binding in regulating the degree of activation of circulating platelets. PMID- 2548626 TI - Photoreactions of human lens monomeric crystallins. AB - Human lens beta s- and gamma A-crystallins exhibit very similar tryptophan fluorescence emission maxima (329 nm). gamma A isolated from infant human lenses is photo-oxidized by 300 nm irradiation and forms water-insoluble aggregates; beta s or gamma A from young human lenses form a small amount of water-soluble crosslinked species. At least part of the mechanism of photodamage by 300 nm irradiation is photogeneration of the oxidant H2O2 via the generation of O2- radical, this reaction occurs via photosensitization by the tryptophan photo oxidation product N-formylkynurenine (N-FK) or related species. These results indicate that even though the tryptophan residues of beta s- and gamma A crystallins are in hydrophobic (buried) microenvironments as compared to those of the alpha- and beta-crystallins, the photogeneration of N-FK is sufficient to produce O2- and H2O2. PMID- 2548627 TI - Lysosomal proteinase-sensitive regions in fast and slow skeletal muscle myosins. AB - We investigated the limited proteolysis of fast and slow myosins purified from rabbit psoas major and semimembranosus proprius muscles, respectively, by the main lysosomal proteinases: cathepsins B, H, L, and D. In EDTA containing buffer, cathepsin D cleaved both myosins only at the rod-S1 junction leading to the formation of two S1 fragments of slightly higher Mr than the three forms obtained with chymotrypsin. On addition of MgCl2 instead of EDTA, myosin hydrolysis was markedly reduced. In contrast, irrespective of the presence of either MgCl2 or EDTA, cathepsin B hydrolysed both myosins into HMM and LMM. Cathepsin L digested myosins more extensively than cathepsins B and D and the main fragments generated were, in decreasing order of importance, rod, S1, S2, HMM, and LMM. In the incubation conditions tested, cathepsin H displayed nondetectable action on myosins. As fast and slow myosin digest patterns were compared, the main differences observed concerned the size of the proteolytic products and the rate of hydrolysis, which was about 4-fold higher for the fast than for the slow isoform. This appeared consistent whatever enzyme was considered. PMID- 2548628 TI - Horizontal organization of orientation-sensitive cells in primate visual cortex. AB - In the visual cortex of the monkey the horizontal organization of the preferred orientations of orientation-selective cells follows two opposing rules: (1) neighbors tend to have similar orientation preferences, and (2) many different orientations are observed in a local region. We have described a classification for orientation maps based on the types of topological singularities and the spacing of these singularities relative to the cytochrome oxidase blobs. Using the orientation drift rate as a measure we have compared simulated orientation maps to published records of horizontal electrode recordings. PMID- 2548629 TI - [Lyophilized membranes as a test system for analyzing the properties of neuromediator receptors and activity of neurotropic agents]. AB - The effect of lyophilization and prolonged storage of rat brain membrane preparations on the properties of receptors of cholinergic and adrenergic neuromediator systems has been studied by the radioreceptor assay. Lyophilized membrane preparations have been shown to be highly stable as compared with fresh membranes and retain their binding properties during storage for 1 year. PMID- 2548630 TI - Neuroendocrine effects of ovine corticotropin-releasing hormone in panic disorder patients. AB - A number of neuroendocrine abnormalities have been reported in panic disorder patients: the most extensively studied being disturbances of hypothalamic pituitary-adrenal function (Curtis et al. 1982; Leiberman et al. 1983; Uhde et al. 1988). The recent sequencing and synthesis of corticotropin-releasing hormone now allows direct testing of pituitary responsivity to this neuropeptide in affective and panic disorder patients (Holsboer et al. 1984; Gold et al. 1986; Roy-Byrne et al. 1986; Holsboer et al. 1987; Risch et al. 1988). We report the effects of intravenously administered ovine corticotropin-releasing hormone (0.03 micrograms/kg) on plasma concentrations of adrenocorticotropin hormone (ACTH) and cortisol in a small group of panic disorder patients and age- and sex-matched normal controls. PMID- 2548631 TI - The corticotropin-releasing hormone test in patients with posttraumatic stress disorder. AB - To evaluate the hypothalamic-pituitary-adrenal (HPA) axis in patients with posttraumatic stress disorder (PTSD), we measured adrenocorticotropin hormone (ACTH) and cortisol responses following administration of corticotropin-releasing hormone (CRH) in 8 combat veterans with chronic PTSD. The PTSD patients had a significantly lower ACTH response to CRH compared to a control group of normal volunteers. Blunted ACTH responses occurred in patients with PTSD alone, as well as those PTSD patients who also had major depression. The cortisol response, although reduced, was not significantly different from normal. The blunted ACTH response to CRH in PTSD patients is similar to that seen in other psychiatric disorders, such as depression, panic disorder, and anorexia nervosa. PMID- 2548632 TI - Estrogen interconversions in the induced cycle in female rhesus monkeys. AB - To determine the extractions and interconversions of estrone and estradiol across and within the uterus, [3H]estradiol and [14C]estrone were infused at a constant rate in six ovariectomized female rhesus (Macaca mulatta) monkeys. Studies were done on Days 9, 14, and 23 of artificial menstrual cycles induced by the timed insertion and removal of Silastic capsules of estradiol and progesterone. Measurements of estrogen radioactivity were made from peripheral arterial blood and uterine venous blood as well as from endometrial biopsy samples. A significant increase occurred in the conversion of estradiol to estrone measured within the uterus on Day 23 compared to Days 9 and 14. The conversion of estrone to estradiol, measured within the uterus, fell progressively from Day 9 to Day 23, but this decrease was not significant. The extractions and interconversions across the uterus, and the overall interconversions of estrone and estradiol were not significantly different on Days 9, 14, or 23 of the cycle. Thus, we have been able to confirm in vivo the increase in the activity of the 17 beta hydroxysteroid dehydrogenase, the enzyme responsible for estradiol to estrone interconversions, shown earlier by studies done in vitro. However, the increase in 17 beta-hydroxysteroid activity in the uterus is not reflected in the overall interconversions of estrone and estradiol as reflected by measurements in peripheral arterial blood. PMID- 2548633 TI - Use-dependent block of single sodium channels by lidocaine in guinea pig ventricular myocytes. AB - Single sodium channel openings have been recorded from cell-attached patches of isolated guinea pig ventricular myocytes. A paired pulse protocol was used to test the hypothesis that channel openings are required for lidocaine block. While the averaged ensemble current during the test pulse was much reduced, there was no correlation between the appearance of channel openings during the conditioning pulse and the subsequent test pulse. Analysis of single channel records demonstrated that the unit conductance of open channels was not changed by lidocaine. The block of ensemble INa was explained by roughly equal reductions in number of open channel events, and in the average duration of opening for each event. These results suggest that lidocaine binding to Na+ channels is dependent upon voltage, but may occur before channel opening. A lidocaine-modified channel can still open, but will be less likely to remain open than a drug-free channel. These results are consistent with block of a pre-open state of the channel. PMID- 2548634 TI - Intracellular magnesium blocks sodium outward currents in a voltage- and dose dependent manner. AB - Tail currents through Na+ channels have been measured in inside-out patches from Xenopus laevis oocytes injected with cDNA-derived mRNA coding for the rat brain type II Na+ channel. It is shown that intracellular Mg2+ blocks outward currents in a voltage- and dose-dependent manner with a half blocking concentration between 3 and 4 mM at 0 mV and a voltage dependence of e-fold per 49 mV. PMID- 2548635 TI - The effects of experimental uremia in rats on duodenal VIP levels and the interaction of VIP with duodenal epithelial cells. AB - The effects of experimental uremia on the concentration of vasoactive intestinal peptide (VIP) in duodenum as well as on the interaction of this neuropeptide with the corresponding epithelial cells were studied in rats. Duodenal VIP concentration was significantly decreased in uremic rats as compared to control animals. The specific binding of VIP to duodenal epithelial cells increased in rats with uremia due to an increase in the number of VIP receptors rather than a change in the binding affinity or in the extent of VIP degradation. On the other hand, the efficacy but not the potency of VIP upon cyclic AMP generation varied in parallel to that observed at the receptor level. PMID- 2548636 TI - Effects of phenoxybenzamine on insulin secretion from isolated rat islets of Langerhans. AB - In isolated rat islets the alpha 2-adrenergic antagonist phenoxybenzamine was found to be only partially effective at relieving the inhibition of glucose induced insulin secretion mediated by noradrenaline. Further experiments revealed a direct inhibitory effect of phenoxybenzamine itself on the secretory response to glucose. At concentrations above 1 microM the antagonist inhibited insulin secretion in a dose-dependent manner, with greater than 50% inhibition at 50 microM. The inhibition of secretion developed rapidly in perifused islets, and was not altered when islets were also incubated with idazoxan or benextramine, suggesting that it did not reflect binding of phenoxybenzamine to the alpha 2 receptor. Paradoxically phenoxybenzamine significantly increased the basal secretion rate in the presence of 4 mM glucose. The results demonstrate that phenoxybenzamine can exert direct effects on insulin secretion which are unrelated to its alpha-antagonist properties. PMID- 2548637 TI - Changes in lipoprotein binding and uptake by hepatocytes during rat liver regeneration. AB - The binding and uptake of cholesterol enriched lipoproteins by isolated hepatocytes was decreased at 16 hours after partial hepatectomy, with a tendency to return to control values as the regeneration proceeds. The number of lipoprotein binding sites of total cellular membranes remained similar to control at 16 and 24 hours. The plasma lipoprotein pattern, determined by electrophoretic analysis, showed a lower per cent of very low density lipoproteins (VLDL) and a higher per cent of low density lipoproteins (LDL) at 16 and 24 hours post-partial hepatectomy. At these times, plasma lecithin: cholesterol acyltransferase (LCAT) activity was decreased. It is intriguing to suggest that the regenerating liver could regulate the blood lipoprotein pattern and the uptake of lipoproteins by modulating the surface expression of the receptors. PMID- 2548638 TI - A comparison of bradykinin, angiotensin II and muscarinic stimulation of cultured bovine adrenal chromaffin cells. AB - Bradykinin, angiotensin II and a muscarinic agonist, acetyl-B-methacholine (methacholine) were all found to elicit catecholamine release from cultured bovine adrenal chromaffin cells. Bradykinin was the most potent of these secretagogues and methacholine the weakest, with angiotensin II intermediate in efficacy. All three secretagogues were much less effective than nicotinic stimulation. The three secretagogues all produced a rise in cytoplasmic free calcium concentration ([Ca2+]i), measured with the fluorescent indicator fura2, which was partially independent of external calcium. In the case of bradykinin the full rise in ([Ca2+]i) may involve a component of calcium entry in addition to release of calcium from an internal store. Secretion was also found to be partially independent of external calcium. The different efficacies of the three secretagogues in eliciting secretion were correlated with the rise in ([Ca2+]i) produced. The differing efficacies of the three secretagogues may be due to the extent of release of calcium from an intracellular store which itself is less effective in eliciting secretion than a rise in [Ca2+]i following calcium entry due to nicotine. Bradykinin also stimulates calcium entry, and this may increase the efficacy of the initial rise in [Ca2+]i. Treatment with pertussis toxin resulted in an enhancement of secretion in response to all of the secretagogues. PMID- 2548639 TI - Inhibition of platelet-von Willebrand factor binding to platelets by adhesion site peptides. AB - Synthetic peptides containing the adhesion site recognition sequences present on the A alpha and gamma chains of fibrinogen were studied for their effect on the binding of endogenous platelet-von Willebrand factor (vWF) and exogenous plasma vWf to thrombin-stimulated platelets. In agreement with previously reported data, the tetrapeptide consisting of the RGDS sequence was a more potent inhibitor of plasma-vWf binding to platelets than was the pentadecapeptide of the carboxy terminus of the fibrinogen gamma-chain (IC50 10.6 mumol/L for the RGDS tetrapeptide v 44.9 mumol/L for the gamma-chain pentadecapeptide). No apparent synergy in the inhibition of plasma-vWf binding was noted when the RGDS and gamma chain peptides were used together (IC50 15.2 mumol/L). In contrast, the gamma chain peptide was significantly more inhibitory than was the RGDS tetrapeptide on the binding of platelet-vWf to platelets (IC50 1.4 mumol/L for the gamma-chain pentadecapeptide v 4.5 mumol/L for the RGDS tetrapeptide, P less than .05), and there was significant synergy in the inhibition of platelet-vWf binding noted when the gamma-chain and RGDS peptides were used together (IC50 0.04 mumol/L). These results indicate that the binding of platelet-vWf to its receptor on the platelet glycoprotein IIb/IIIa complex involves both the RGDS and gamma-chain recognition sites. In contrast to the results with plasma-vWf binding, the gamma chain recognition site appears to be more important than the RGDS recognition site in platelet-vWf binding to platelets. PMID- 2548640 TI - Functional characteristics of receptor-bound urokinase on human monocytes: catalytic efficiency and susceptibility to inactivation by plasminogen activator inhibitors. AB - We compared urokinase-type plasminogen activator (u-PA) in fluid phase and u-PA bound with its receptor on human blood monocytes with respect to proteolytic activity and susceptibility to inactivation by the plasminogen activator inhibitors PAI-1 and PAI-2. Receptor-bound u-PA is catalytically twice as efficient as fluid-phase u-PA. Fluid-phase u-PA is susceptible to rapid inhibition by PAI-1 and PAI-2 at an estimated PAI:u-PA molar ratio of 2:1. In contrast, u-PA bound to endogenously occupied receptors is inhibited by PAI-2 only at PAI:u-PA molar ratios of 20:1, but is not inhibited by PAI-1, u-PA/PAI-1 and u-PA/PAI-2 complexes bind to the receptor with a tenfold lower affinity than u-PA itself. Thus, competition of u-PA/PAI complexes with fluid-phase u-PA for binding to the receptor is unlikely to affect the overall plasminogen activator activity of the monocyte. These findings demonstrate that the activity of receptor-bound u-PA can be modulated by PAI-2, but not by PAI-1, to adjust the cell's proteolytic activity to different local situations. PMID- 2548641 TI - Herpes zoster infection after autologous bone marrow transplantation. AB - One hundred fifty-three patients who underwent autologous bone marrow transplant (ABMT) were studied retrospectively to determine the frequency, outcome, and risk factors associated with varicella-zoster infections (VZV). Forty-three patients (28%) developed VZV infection after transplant. The median onset of infection was the fifth month, with 91% of cases occurring within the first year. Thirty-three patients (77%) had localized herpes zoster, and ten patients (23%) had varicella. Cutaneous dissemination developed in 15% of patients and probable visceral dissemination developed in 5%. Overall morbidity was 25% and included scarring, alopecia, postherpetic neuralgia, and neurologic dysfunction. There were no deaths from VZV infection. The majority of patients (79%) were treated with intravenous (IV) acyclovir. The only significant risk factor associated with VZV infection was the underlying disease. VZV infection occurred most frequently in patients with Hodgkin's and non-Hodgkin's lymphoma (46%) as compared with patients with leukemia (23%) or solid tumors (9%) (P less than .002). The frequency of VZV infection in ABMT patients appears to be comparable to that reported for allogeneic BMT patients and other immunocompromised patients. PMID- 2548642 TI - Suppression of natural killer cell activity by FG 7142, a benzodiazepine receptor "inverse agonist". AB - A dose-dependent (5-50 mg/kg) suppression of natural killer (NK) cell activity was observed 2 h after administration of the benzodiazepine receptor "inverse agonist" FG 7142 (N-methyl-beta-carboline-3-carboxamide), and was still manifest 24 h later. Addition of FG 7142 (1-1000 nM) to the 4 h 51 Cr release assay did not affect NK cell activity. Pretreatment of mice with the benzodiazepine receptor antagonist Ro 15-1788 (10 mg/kg) blocked FG 7142-induced suppression of NK cell activity, but had no effect when administered alone. The suppression of NK cell activity by FG 7142, a compound which produces a syndrome resembling stress or anxiety in both animals and man, provides further evidence that the central nervous system pathways subserved by the benzodiazepine/GABA receptor chloride channel complex ("supramolecular complex") may play a role in the modulation of immune function. PMID- 2548643 TI - Purification of rat liver soluble catechol-O-methyltransferase by high performance liquid chromatography. AB - The soluble form of catechol-O-methyltransferase (EC 2.1.1.6) from rat liver was purified to homogeneity by high-performance anion-exchange chromatography and high-performance gel-filtration chromatography. The specific activity of the final pool was 270 U/mg protein. The purification was 1180-fold and recovery of the enzyme activity was 15%. During this rapid and gentle purification there were no problems with loss of activity, and the estimated half of the final purified enzyme pool was 5.5 days at +4 degrees C. The only additive used was phenylmethylsulfonylfluoride in the homogenizing buffer. PMID- 2548644 TI - Identification of point of specific enzymic cleavage of P1,P4-bis(5'-adenosyl) tetraphosphate by negative ion FAB mass spectrometry. PMID- 2548645 TI - Human PTH(1-34) infusion test in differential diagnosis of various types of hypoparathyroidism: an attempt to establish a standard clinical test. AB - To introduce a simple procedure and reliable diagnostic criteria for parathyroid hormone (PTH) infusion test, 128 patients with either pseudo- (PsH) or idiopathic hypoparathyroidism (IdH) and 25 normocalcemic controls were studied. Incremental responses of urinary cyclic AMP and phosphate to 20 micrograms (67 U) or 30 micrograms (100 U) of human PTH(1-34) were assessed by using simple parameters of urinary excretion rates of the two substances. The results are summarized as follows. (1) PTH dose-cyclic AMP response relation suggests that 100 U of PTH is more appropriate than 67 U as a standard test dose for adults. (2) By presenting the magnitude of cyclic AMP response as either net increase or fold increase during 1 h after 100 U of PTH infusion, we can differentiate PsH type I from others without overlap. (3) Differentiation between PsH and IdH or normocalcemic subjects by phosphaturic response is less clearcut than that made by cyclic AMP response whatever indices and criteria are used. Thus it seems difficult to diagnose PsH type II merely based on the discrepancy between cyclic AMP and phosphaturic responses to exogenous PTH. (4) The test results are essentially similar in the examinations performed before and during vitamin D therapy. However, when the magnitude of phosphaturic response is expressed as net increase during 2 h after PTH, it tends to be enhanced after vitamin D therapy in patients with PsH compared to the response before therapy. PMID- 2548646 TI - The adenylate cyclase response to parathyroid hormone in cultured rabbit marrow fibroblastic cells. AB - The ability of fibroblastic cells to respond to parathyroid hormone (PTH) by an increase in adenylate cyclase activity is accepted as a characteristic of the osteogenic phenotype. Whether marrow fibroblastic cells, which have osteogenic potential when assayed in vivo, demonstrate this hormonal response when cultured in vitro has been investigated. Our study has shown a level of stimulation of adenylate cyclase activity by PTH in cultured rabbit marrow fibroblasts comparable with other osteogenic cells in vitro. The effect is seen in fibroblasts grown either from multiple colonies or from single colonies. Only a proportion of colonies had osteogenic potential in vivo assay and our results show a similar finding for the PTH response in vitro. To what degree the two parameters are expressed by the same colony has not yet been established. PMID- 2548647 TI - Progressive neurological dysfunction during latent HIV infection. AB - OBJECTIVE--To determine whether the delayed conduction through the spinal cord and peripheral nerves seen in patients with AIDS is related to infection with HIV or to the presence of an immunodeficient state. DESIGN--Two year prospective follow up study of electrophysiological measurements in subjects positive for HIV antibody but without AIDS. SETTING--HIV screening clinic and clinical departments in a university hospital in Copenhagen, Denmark. SUBJECTS--Twelve homosexual men positive for HIV antibody who had not developed AIDS. RESULTS--Eight latencies were measured: from the ankle to T12, the wrist to C7, T12 to the cerebral cortex, C7 to the cerebral cortex, the ankle to the gluteal crease (tibial nerve), the gluteal crease to T12, the wrist to Erb's point (median nerve), and Erb's point to C7. Spinal latencies increased in all subjects at C7 by a mean of 4.2% (SE 0.9%) and in all except one at T12 by a mean of 5.5% (1.0%). The conduction time from the gluteal crease to T12 was increased by a mean of 32.0% (5.0%) whereas that in the median and tibial nerves by only 5.6% (1.0%) and 2.2% (2.2%) respectively. CONCLUSIONS--A mild and slowly progressive peripheral neuropathy of the axonal type and a more severe progressive myelopathy or myeloradiculopathy occur concomitantly with early HIV infection, possibly as the result of a direct neurotropic action of HIV. PMID- 2548648 TI - Recent advances in the treatment of small cell lung cancer. AB - Small cell lung cancer is a rapidly dividing tumour that is susceptible to combination chemotherapy and radiotherapy. Improvements in treatment regimens have resulted in better response rates, but long-term survival has not improved over the past decade. PMID- 2548649 TI - Modern cardiac pacing: the role of dual chamber systems. AB - Atrial synchronous or dual chamber pacemakers are at the forefront of current pacing technology. This article discusses the mechanisms of dual chamber pacing and considers the role of such systems in modern clinical practice. PMID- 2548650 TI - Ductal carcinoma in situ: a new clinical problem. PMID- 2548651 TI - Follow-up of two treatment modalities for ductal cancer in situ of the breast. AB - In a mammography screening programme for the early detection of breast cancer 66 cases of ductal cancer in situ of the breast (DCIS) were diagnosed between 1978 and 1984 and prospectively followed up. In the beginning of the study period, DCIS was treated by mastectomy without axillary clearance but sector resection has been performed increasingly. Since 1982 the latter procedure has become standard treatment. Radical resection was confirmed by specimen X-ray and histopathological examination of whole organ sections. No postoperative radiotherapy was given. Twenty-eight women had mastectomy and 38 had sector resection. The median follow-up times were 77 and 60 months respectively. In the mastectomy group postoperative monitoring did not reveal any local recurrence but one contralateral new invasive cancer was discovered. In the sector resection group five recurrences were found. Three of the latter were new cases of DCIS and two appeared as small invasive cancers (stage I). Mastectomy will cure ductal cancer in situ but has a greater morbidity. Sector resection has been established as the method of choice in stage I invasive cancer and is probably also safe in DCIS. The possible beneficial effect of postoperative local radiotherapy after sector resection for DCIS is currently being analysed in a randomized trial which started in Sweden in 1987. PMID- 2548652 TI - Intraoperative immunostaining for detection of invasive cells at the resection margin of Borrmann type 4 gastric carcinoma using monoclonal antibody S202. AB - Borrmann type 4 gastric cancer is clinically characterized by diffuse infiltration with carcinoma cells. An antiscirrhous carcinoma monoclonal antibody (MAb) S202 was used in a rapid immunostaining procedure to identify the limit of tumour invasion during surgery in samples obtained from the resection margins in 37 cases of Borrmann type 4 gastric cancer. In two instances, conventional cytological diagnosis using haematoxylin and eosin staining was negative, and in another it was inconclusive; whereas by the rapid immunostaining method single cells stained darkly, indicating malignancy. In two specimens, a positive diagnosis could be made by both methods, but immunostaining of these sections served to highlight the adenocarcinoma cells against the inflammatory background: further resection was performed. This technique enables clear recognition of the infiltrating tumour cells in the frozen sections of resected specimens. The rapid immunostaining method using MAb S202 allowed accurate and rapid determination of the limit of tumour extension at the surgical margin during surgery. PMID- 2548653 TI - Familial polyposis: colon cancer in the absence of rectal polyps. PMID- 2548654 TI - Juvenile polyp in an ileoanal J pouch following restorative proctocolectomy for juvenile polyposis coli. PMID- 2548655 TI - Endoscopic management of inoperable cholangiocarcinoma using iridium-192. AB - We report a well tolerated endoscopic technique of administering intraluminal radiotherapy to 14 patients with inoperable cholangiocarcinoma, in which the iridium-192 wire source was inserted down a nasobiliary catheter placed within a previously inserted endoscopic biliary prosthesis, thus allowing bile flow to continue during treatment. Radiotherapy was commenced 2 weeks after biliary decompression, when the median serum bilirubin level had fallen from 213 to 34 mumol/l. A total radiation dose of 6000 cGy at 0.5 cm from the source was administered over a median of 85 h (range 77-116 h). In four patients there was a transient increase in serum bilirubin during iridium treatment and, in two cases (14 per cent), this was associated with mild cholangitis. Both cases resolved rapidly once the iridium wire and nasobiliary catheter were removed. The median hospital stay after treatment was 2.5 days (range 0-28 days). The late complications associated with stent blockage were minimized by routinely changing stents at 4-6 monthly intervals; ten of the patients have so far undergone from one to five (median three) stent changes. Five patients have died at 3.6-8.2 months (median 4.8 months) following iridium treatment; four had extensive type III hilar lesions at diagnosis. Nine patients are still alive at 5.4-31.0 months (median 16.4 months). The overall median survival after iridium therapy is 10.5 months. This technique of endoscopic biliary decompression before internal iridium administration minimizes complications by allowing biliary drainage during treatment. The role of this treatment in patients with inoperable cholangiocarcinoma awaits the results of randomized, controlled trials. PMID- 2548657 TI - Coupling between hypothalamic alpha 2-adrenoceptors and [3H]mazindol binding site in response to several hyperglycaemic stimuli in mice. AB - The hypothalamic response to circulating glucose and insulin levels was studied in the mouse by differentially attenuating glucose-homeostasis. The administration of glucose, 2-deoxyglucose or the alpha 2-adrenoceptor agonist UK 14.304 was accompanied by a persistent hyperglycaemia; however, an increase in insulin levels was obtained with glucose and a decrease with the other two manipulations. Both alpha 2-adrenoceptors (labeled with [3H]idazoxan) and the anorectic recognition site (labeled with [3H]mazindol) were upregulated by the three treatments. A good correlation was obtained between circulating glucose levels and either hypothalamic [3H]mazindol binding (r = 0.70, P less than 0.001) or [3H]idazoxan binding (r = 0.63, P less than 0.001), as well as between the two binding sites (r = 0.88, P less than 0.001). No correlation was obtained between circulating insulin levels and these binding sites (r = 0.18, r = 0.26, P = n.s. for [3H]mazindol and [3H]idazoxan binding, respectively). It is suggested the alpha 2-adrenoceptors and the anorectic binding sites are associated in their response to glucose as part of a hypothalamic center involved in the regulation of feeding mechanisms. PMID- 2548656 TI - Astrocyte morphology altered by 1-(5-isoquinolinylsulfonyl) 2-methyl piperazine (H-7) and other protein kinase inhibitors. AB - Studies were conducted to determine if the protein kinase C inhibitor H-7 could block the effects of phorbol-12-myristate-13-acetate (PMA) on astrocyte morphology. Contrary to expectation, H-7 alone was found to induce morphological changes very similar to those elicited by PMA. This effect was shared by two other inhibitors of protein kinase C, H-8 and staurosporine, but not by the cyclic nucleotide-dependent protein kinase inhibitor HA-1004 or the calcium/calmodulin dependent protein kinase inhibitor W-7. Although the morphological effects observed with H-7 resemble those induced by PMA, H-7 did not promote the redistribution of protein kinase C to the membrane or induce the phosphorylation of endogenous proteins like PMA. In addition, the effects of H-7 were still observed in cells depleted of protein kinase C activity which were no longer responsive to treatment with PMA. Cytoskeletal elements appear to be involved in the effect of H-7 on cell shape since this effect is blocked by treatment with colchicine. Activators of the cyclic AMP-dependent protein kinase also alter astrocyte shape, however, while H-7 did cause a slight increase in cyclic AMP levels, it was unlikely that this action is responsible for its effect on morphology. One common action of both H-7 and PMA was to decrease the 32P content of several 20,000 Da proteins. While the mechanism by which H-7 exerts its influence on astrocyte morphology remains to be clarified, be it by the inhibition of protein kinase C or some other mechanism, the results suggest that caution must be used when interpreting the effects of activators and inhibitors of this kinase. PMID- 2548658 TI - Nerve growth factor receptor-mediated transport from CSF labels cholinergic neurons: direct demonstration by a double-labeling study. AB - It has recently been shown that, following intraventricular administration, a monoclonal antibody directed against rat nerve growth factor (NGF) receptor is specifically accumulated bilaterally by numerous neurons of the basal forebrain. The labeled neurons have a morphology and topography which are characteristic of the magnocellular cholinergic basal forebrain (CBF) system. This is also consistent with the evidence that CBF neurons have NGF receptors and respond to NGF in a variety of experimental situations. In the present report, a double labeling technique is used to directly demonstrate that choline acetyltransferase containing (and therefore cholinergic) neurons do in fact accumulate the anti-NGF receptor antibody from CSF. PMID- 2548659 TI - In vivo labeling of central benzodiazepine receptors with the partial inverse agonist [3H]Ro 15-4513. AB - Ro 15-4513 is an imidazobenzodiazepine and a partial inverse agonist at the central benzodiazepine receptors (BZDr). It has been shown to antagonize behavioral and biochemical effects of ethanol. In vivo binding of [3H]Ro 15-4513 was evaluated in mouse brain. After intravenous injection [3H]Ro 15-4513 was readily taken up by the brain and distributed to brain areas enriched in benzodiazepine receptors. Binding was specific for central BZDr, saturable and reversible. A high degree of specific binding, relative to non-specific binding, was achieved. Analysis of dissociation kinetics revealed that [3H]Ro 15-4513 was retained significantly longer in hippocampus compared to other brain regions. In view of the known distribution of benzodiazepine receptor subtypes, this suggests that, in vivo, [3H]Ro 15-4513 has a higher affinity for benzodiazepine receptors type II and may explain quantitative differences in the regional distribution of this ligand compared to the antagonist [3H]Ro 15-1788. We conclude from these studies that Ro 15-4513 is a suitable ligand for in vivo studies of benzodiazepine receptors. Labeled with a positron-emitting isotope, it could be used with positron emission tomography to study BZDr in man under a variety of conditions. PMID- 2548660 TI - Inhibition by sodium valproate of the arginine vasopressin and adrenocorticotropin responses to angiotensin II in normal men. AB - The present study was carried out in order to determine whether angiotensin II (ANG II) exerts its stimulatory effect on plasma concentration of arginine vasopressin (AVP) and adrenocorticotrophic hormone (ACTH) by interacting with a (gamma-aminobutyric acid) GABAergic pathway. For this purpose, the effect of the GABAergic agonist sodium valproate (600 mg in 3 divided doses p.o.) on the ACTH and AVP responses to ANG II (infusion for 60 min of successively increasing doses of 4, 8 and 16 ng/kg min; each dose for 20 min) administration was evaluated in 7 normal men. In all subjects control ANG II tests were performed with a placebo instead of sodium valproate. ANG II induced a significant increase of AVP at 60 min and ACTH at 90 min after the beginning of infusion. The pretreatment with sodium valproate reduced significantly both ANG II-induced AVP and ACTH rises, without changing the time course of the hormonal responses. These data suggest the involvement of a GABAergic mechanism in regulation of both AVP and ACTH responses to ANG II in man. PMID- 2548661 TI - Two distinct quisqualate receptor systems are present on striatal neurons. AB - At concentrations at which it did not alter spontaneous release, quisqualate (QUIS) induced a dose-dependent (EC50, 0.5 microM) potentiation of KCl- or veratrine-evoked release of [3H]GABA from striatal neurons in primary culture. QUIS potentiation of KCl-evoked [3H]GABA release was mimicked by the selective agonist alpha-amino-3-hydroxy-5-methylisoxazole-propionic acid (AMPA), glutamate and kainate, and was blocked by kynurenic acid and gamma-D-glutamylglycine. QUIS also induced a dose-dependent (EC50, 0.2 microM) augmentation of [3H]inositol monophosphate production in striatal neurons. This action of QUIS was mimicked by glutamate, but not by AMPA nor by kainate. Furthermore, none of the antagonists tested (kynurenic acid, gamma-D-glutamylglycine, glutamic acid diethyl ester, and 4-aminophosphonobutanoic acid) could block QUIS-induced elevations in [3H]inositol monophosphate production. The results of the present study suggest that two QUIS receptor systems, distinguished on the basis of their pharmacological properties, may subserve specific roles in the regulation of striatal neuron function by excitatory amino acids. PMID- 2548662 TI - Chronic haloperidol administration increases GABA binding and enhances neuronal responsiveness to iontophoresed GABA in rat globus pallidus. AB - Experiments were conducted to assess the effects of chronic haloperidol (CHAL) administration on gamma-aminobutyric acid (GABA) receptor binding within the rat globus pallidus (GP) and on the responsiveness of individual pallidal neurons to microiontophoretically applied GABA and glycine. Rats were administered haloperidol in their food for 30 days in increasing concentrations and the experiments were conducted 2 days after termination of the haloperidol treatment. GABA receptor binding and neuronal responsiveness to GABA were significantly increased within the GP following CHAL treatment. The mean EC50 value for GABA was significantly decreased in the CHAL-treated rats, but there was no change in the EC50 for glycine. Scatchard analysis of [3H]muscimol binding demonstrated a single high-affinity binding site (Kd = 5 nM) within both control and CHAL treated rats. The binding capacity (Bmax) of this high-affinity site was significantly increased in CHAL-treated rats without any change in the dissociation constant (Kd) for this site. These results suggest that CHAL administration may lead to a decrease in GABA release by striatopallidal efferents. The results of this study, combined with those of our previous study on SNR neurons, have demonstrated that blockade of striatally mediated dopamine (DA) neurotransmission leads to similar changes in GABAergic mechanisms at the level of the GP and SNR and suggest that DA regulation of the striatopallidal and striatonigral GABAergic pathways need not be differentially organized as has previously been postulated. PMID- 2548663 TI - Dual opioid modulation of the action potential duration of mouse dorsal root ganglion neurons in culture. AB - Multiple modulatory effects of opioids on the duration of the calcium component of the action potential (APD) of dorsal-root ganglion (DRG) neurons of mouse spinal cord-ganglion explants were studied. The APD of DRG neuron perikarya has been previously shown to be shortened by exposure to high concentrations of opioids (ca. 0.1-1 microM) in about 1/2 of the cells tested. The present study demonstrates that in addition to these inhibitory modulatory effects of opioids, lower concentrations (1-10 nM) of present study demonstrates that in addition to these inhibitory modulatory effects of opioids, lower concentration (1-10 nM) of delta- mu, and kappa-opioid agonists elicit excitatory modulatory effects, i.e. prolongation of the APD, in about 2/3 of the sensory neurons tested. APD prolongation as well as shortening elicited by delta, mu, and kappa agonists were prevented by coperfusion with the opioid antagonists, naloxone or diprenorphine (10 nM). APD prolongation induced by the delta-agonist [D-Ala2-D-Leu5]enkephalin (DADLE) was prevented in the presence of multiple K+ channel blockers, whereas excitatory modulation by the specific kappa-agonist, U-50,488H was not attenuated under these conditions. After treatment of DRG neurons with pertussis toxin (1 micrograms/ml for several days) or forskolin (50 muM for less than 15 min), a much smaller fraction of cells showed opioid-induced APD shortening; moreover, a much larger fraction of cells showed opioid-induced APD prolongation, even when tested with high concentrations of DADLE (1-10 muM). These data indicate that opioid-induced APD prolongation is not mediated by pertussis toxin-sensitive G proteins (which have been shown to regulate opioid inhibitory effects) and suggest that elevation of cyclic AMP levels may enhance opioid excitatory responsiveness. Furthermore, our analyses indicate that mu-, delta- and kappa subtypes of excitatory as well as inhibitory opioid receptors may be expressed on the same DRG neuron perikaryon under in vitro conditions. If dual opioid modulation of the APD of DRG perikarya also occurs in central DRG terminals this may play a significant role both in nociceptive signal transmission as well as tolerance to opioid analgesia. PMID- 2548664 TI - Production of immunoreactive adrenocorticotropin and beta-endorphin by hypothalamic and extrahypothalamic brain cells. AB - Despite many in vivo studies, little is known about brain regulation of POMC synthesis or regulation of secretion of POMC-related peptides. To test the hypothesis that dissociated brain cells in culture can produce and release POMC related peptides, immunoreactive (IR)-adrenocorticotropin (ACTH) and beta endorphin were measured in cells and media of dissociated cell cultures incubated up to 38 days. Fetal rat hypothalamic and extrahypothalamic forebrain cells were maintained in serum free medium. IR-ACTH and beta-endorphin were measured by radioimmunoassay in concentrated cells and media after various incubation times using two ACTH (mid-portion = R4; carboxy-portion directed = KEND) antisera and a beta-endorphin antiserum. IR-ACTH and IR-beta-endorphin in hypothalamic and extrahypothalamic cells and in media (cumulative) were greater than quantities in cells before culture. Peak hypothalamic cellular content of IR-ACTH (5.3 fmol/10(6) cells-R4; 4.7 fmol/10(6) cells-KEND) and content of IR-beta-endorphin (32.0 fmol/10(6) cells) occurred on days 16, 9 and 23, respectively. Peak extrahypothalamic content of IR-ACTH (2.9 fmol/10(6) cells-R4; 1.0 fmol/10(6) cells-KEND) and content of IR-beta-endorphin (10.8 fmol/10(6) cells) was also seen on different days, was lower than hypothalamic content and was not always concurrent with peak hypothalamic content. Gel filtration chromatography revealed that the predominant forms of IR-ACTH and IR-beta-endorphin in hypothalamic cell extracts co-eluted with synthetic ACTH1-39 and beta-endorphin. Changes in molar ratios of IR-ACTH and IR-beta-endorphin also suggested a differential regulation of different POMC derivatives. CONCLUSIONS: (1) IR-ACTH and IR-beta-endorphin are produced by hypothalamic and extrahypothalamic forebrain cells in culture: and (2) dissociated brain cell cultures can be used as a potential model for studying regulation of POMC-related peptides in brain. PMID- 2548665 TI - CNS cell groups regulating the sympathetic outflow to adrenal gland as revealed by transneuronal cell body labeling with pseudorabies virus. AB - The CNS cell groups that innervate the sympathoadrenal preganglionic neurons of rats were identified by a transneuronal viral cell body labeling technique combined with neurotransmitter immunohistochemistry. Pseudorabies virus was injected into the adrenal gland. This resulted in retrograde viral infections of the ipsilateral sympathetic preganglionic neurons (T4-T13) and caused retrograde transneuronal cell body infections in 5 areas of the brain: the caudal raphe nuclei, ventromedial medulla, rostral ventrolateral medulla, A5 cell group, and paraventricular hypothalamic nucleus (PVH). In the spinal cord, the segmental distribution of virally infected neurons was the same as the retrograde cell body labeling observed following Fluoro-gold injections in the adrenal gland except there was almost a 300% increase in the number of cells labeled and a shift in cell group distribution. These results imply there are local interneurons that regulate the sympathoadrenal preganglionic neurons. In the medulla oblongata, serotonin (5-HT)-, substance P (SP)-, thyrotropin-releasing hormone-, Met enkephalin-, and somatostatin-immunoreactive neurons of the raphe pallidus and raphe obscurus nuclei and the ventromedial medulla were infected. In the ventromedial and rostral ventrolateral medulla, immunoreactive phenylethanolamine N-methyltransferase, SP, neuropeptide Y, somatostatin, and enkephalin neurons were infected. The A5 noradrenergic cells were labeled, as were some somatostatin immunoreactive neurons in this area. In the were infected. The A5 noradrenergic cells were labeled, as were some somatostatin-immunoreactive neurons in this area. In the hypothalamus, tyrosine hydroxylase- and SP-immunoreactive neurons of the dorsal parvocellular PVH were infected. Only a few immunoreactive vasopressin, oxytocin, Met-enkephalin, neurotensin, and somatostatin PVH neurons were labeled. PMID- 2548666 TI - The effects of 4-aminopyridine on the cat spinal cord: rhythmic antidromic discharges recorded from the dorsal roots. AB - In a previous paper, we have reported that 4-aminopyridine (4-AP, i.v., 10 mg/kg) induces in decerebrate spinal and paralyzed cats, a sustained rhythmic activity (2.5-8.5 Hz) in various muscle nerves. We describe here that similar discharges are recorded from the proximal stump of cut cutaneous nerves. The latter rhythmic activity arises from intense antidromic discharges in the dorsal roots. The rhythmic discharges are recorded from dorsal roots of both spinal cord enlargements as well as from thoracic roots. The rhythmic activity is highly synchronous among adjacent dorsal roots. Bilateral activity is also highly cross correlated, but may be dissociated by unilateral stimulation of one dorsal root. It is not yet possible to determine the precise site where the antidromic discharges recorded from the dorsal roots are generated. 4-AP could act directly at the terminal level of the primary afferents or could activate interneurons impinging upon the terminals. PMID- 2548667 TI - Unitary discharges in dorsal and ventral roots after the administration of 4 aminopyridine in the cat. AB - The administration of 4-aminopyridine (4-AP) in decerebrate paralyzed cats induces centrifugal rhythmic discharges in both ventral and dorsal roots. This study describes the mode of discharge of individual primary afferents as well as some ventral root fibers. Several patterns of antidromic discharge have been observed in primary afferents after the administration of 4-AP. A large proportion of the units (n = 96; 53%) showed rhythmic bursts of discharge related (n = 41) or not (n = 55) to the ongoing rhythmic activity in the peripheral nerves. Other units (n = 86; 47%) discharged either tonically, sporadically or had no antidromic activity at all. The conduction velocity of the non-bursting units was significantly higher (89.7 +/- 18.4 m/s) than that of the bursting units (70.6 +/- 15.4 m/s; P less than 0.01). Ventral roots showed rhythmic activity although less intense than that of the dorsal roots. As in dorsal roots, some fibers showed a rhythmical pattern of discharge related to the mass activity recorded from whole dorsal roots or peripheral nerves, while other units were not related. It is concluded that bursting activity which occurs in peripheral nerves after the administration of 4-AP is mainly the result of the antidromic activation of medium to small size primary afferent fibers. PMID- 2548668 TI - GABAA/benzodiazepine receptor-like immunoreactivity in rat and monkey cerebellum. AB - The cellular and subcellular localization of GABAA/benzodiazepine receptor-like immunoreactivity in the rat and monkey cerebellum has been studied with a monoclonal antibody (E9) directed against the alpha-subunit of purified GABAA/benzodiazepine receptors. At both the light and electron microscopic level E9 immunoreactivity is located in all 3 layers of the cerebellar cortex and within the deep cerebellar nuclei. The reaction product accumulates within the cytoplasm of neurons and their dendrites but axons are not immunoreactive. Glial cells in the white matter and the cortical layers are also unlabeled, although in some instances Bergmann glia do contain reaction product. The overall distribution and cellular and subcellular localization of E9 immunoreactivity is identical for both monkey and rat cerebellum. On the basis of cell size, morphology, and location it is evident that E9 immunoreactivity occurs in examples of all 5 neuronal types in the cerebellar cortex: Purkinje cells, Golgi type II cells, granule cells, and stellate and basket cells. However, the distribution of the reaction product within the cells is more selective. For example, electron microscopy demonstrates that axonal processes and terminals are not E9 immunoreactive with the single exception of the mossy fiber terminals in the granular layer. Also, examples of unlabeled axon terminals resembling those derived from Golgi type II cells, basket cells, and stellate cells form synapses with immunoreactive dendrites and cell bodies in the cortical layers. Finally, in the deep cerebellar nuclei unreactive axon terminals make symmetric synapses with immunostained neurons and dendrites. These results show that E9 monoclonal antibodies label neurons and portions of their processes which are postsynaptic in GABA-mediated inhibitory circuits, and demonstrates that this antiserum can be used as a morphological marker for cells which make GABAA/benzodiazepine receptors. PMID- 2548669 TI - Inhibition of cyclic AMP formation in N1E-115 neuroblastoma cells is mediated by a non-cardiac M2 muscarinic receptor subtype. AB - The cardioselective muscarinic antagonist, AF-DX 116 [11[2-[(diethyl-amino) methyl]-O-1-piperidinyl]-5,11-dihydro-6H-pyrido- [2,3-b][1,4]-benzodiazepine-6 one), was weak at blocking the M2 muscarinic receptor-mediated inhibition of cyclic adenosine monophosphate (cAMP) formation in mouse neuroblastoma cells (clone N1E-115). In contrast, the glandular-selective antagonists, hexahydro-sila difenidol (HHSiD) and 4-diphenylacetoxy-N-methyl-piperidine methiodide (4-DAMP), were quite potent at inhibiting this response, being 14- and 318-fold more potent than AF-DX 116 in this regard, respectively. According to the rank order of potency of these two classes of antagonists, these data provide the first pharmacological evidence that inhibition of cAMP formation in a neuronal tissue is mediated by a non-cardiac M2 muscarinic receptor subtype. PMID- 2548670 TI - Developmentally different onset of alpha 1- and alpha 2-adrenergic responses in the neonatal rat dorsal motor nucleus of the vagus in vitro. AB - Development of noradrenergic responses in the rat dorsal motor nucleus of the vagus (DMV) from postnatal days 0 to 22 was studied by a conventional microelectrode technique using brain slice preparations. Between postnatal days 0 and 6, noradrenaline (NA) caused only alpha 1-adrenoceptor-mediated depolarizations. By postnatal day 14, alpha 2-adrenoceptor-mediated hyperpolarization appeared. During the next postnatal week, the response pattern to NA became similar to that in adults, including both alpha 1- and alpha 2 adrenergic responses. These results suggest that alpha 2-adrenergic responses in DMV neurons are absent at birth but develop rapidly in the 3 weeks after birth. Furthermore, alpha 1-adrenoceptors have already been functioning in the fetal period. PMID- 2548671 TI - Effect of ammonium chloride on the astrocyte benzodiazepine receptor. AB - We have carried out studies on the effect of ammonium chloride on the astrocyte benzodiazepine receptor. Scatchard analysis of the binding of [3H]Ro-5-4864 to homogenates prepared from primary astrocyte cultures showed a significant decrease in Kd (27% with 2 mM NH4Cl; 32% with 5 mM NH4Cl; 25% with 10 mM NH4Cl) and Bmax (14% with 10 mM NH4Cl). These findings indicate that ammonium chloride can affect the astrocyte benzodiazepine receptor, and that such receptor changes may contribute to ammonia-induced encephalopathy. PMID- 2548672 TI - Ultrastructural and functional evidence for the survival of corticogeniculate neurons in kainic acid-lesioned lateral geniculate nucleus. AB - After a kainic acid lesion in the dorsal lateral geniculate nucleus of rat, retrograde axonal transport of fluorescent dyes is blocked in corticogeniculate but not in retinogeniculate neurons. This inhibition, however, can be reversed by electrical stimulation in the subcortical white matter (Woodward and Coull, Brain Research 454 (1988) 106-115). These observations suggest that retrograde axonal transport in corticogeniculate neurons is impulse-dependent and that neuronal activity in this pathway is reduced as a consequence of the lesions. To test this we examined retrograde transport of horseradish peroxidase (HRP) and cytochrome oxidase activity in the cortex of lesioned animals. Unilateral kainic acid lesions in the geniculate inhibit the retrograde transport of HRP, but this inhibition is reversed by electrical stimulation of white matter. Moreover, histochemical staining for cytochrome oxidase activity is less intense over visual cortex on the lesioned side, implying that cortical activity in intrinsic and efferent pathways is reduced as a consequence of removal of geniculate afferents. Inasmuch as the retrograde transport of HRP is dependent upon impulse activity in neurons and is thought to be mediated by synaptic vesicle recycling, these results suggest that terminals of corticogeniculate fibers survive the kainic acid lesions in the geniculate and are capable of releasing synaptic vesicles. Ultrastructural examination of lesioned geniculates strongly supports this conclusion and reveals the presence of axon terminal profiles which are filled with small round synaptic vesicles and have membrane specializations reminiscent of synaptic contacts. These terminal profiles are presumed to be of retinal and cortical origin. PMID- 2548673 TI - Three types of voltage-dependent calcium current in cultured rat hippocampal neurons. AB - Voltage-dependent calcium (Ca2+) currents in cultured rat hippocampal neurons were studied with the whole-cell recording mode of the patch-clamp technique. On the basis of the voltage-dependence of activation, kinetics of inactivation and pharmacology, 3 types of Ca2+ currents were distinguished. The low-threshold Ca2+ current (Il) was activated at -60 mV, and completely inactivated during a 100-ms depolarization to -40 mV (time constant: tau = 16 +/- 1 ms). The high-threshold currents (Ih), which were activated at -20 mV, could be separated into two types. The high-threshold, fast inactivating current (Ih,f) decayed quickly during a maintained depolarization (tau = 33 +/- 3 ms at 0 mV), whereas the high threshold, slowly inactivating current (Ih,s) decayed with a much slower time constant (tau = 505 +/- 42 ms at 0 mV). The inactivations of Ih,f and Ih,s exhibited different time- and voltage-dependencies. Nickel ions (Ni2+, 25 microM) markedly suppressed Il, but little affected Ih. Cadmium ions (Cd2+, 10 microM) almost completely suppressed Ih, but left a small amount of Il. Lanthanum ions (La3+, 10 microM) almost completely suppressed both Il and Ih. Ih,s was sensitive to block by the dihydropyridine antagonist nicardipine (10 microM). PMID- 2548674 TI - Opiate receptor subtype binding in gerbil hippocampus is altered by forebrain ischemia. AB - A role for endogenous opioids in trauma-induced brain injury has been supported by pharmacological studies. The present series of experiments were initiated to extend these observations by measuring opiate receptor subtype binding in gerbil hippocampus following 7 days recovery from a 10 min ischemic insult. Quantitative in vitro autoradiography was utilized to measure mu [( 3H]DAGO), kappa [( 3H]bremazocine + 10 microM morphiceptin + 100 nM DSLET), delta [( 3H]DSLET + 10 microM morphiceptin) and lambda [( 3H]naloxone + 300 nM diprenorphine) binding. While ischemic tissue samples at the level of the dorsal hippocampus showed complete loss of CA1 pyramidal cells, we observed no significant alterations in mu or delta binding suggesting a non-pyramidal cell localization of these receptors. Kappa binding decreased significantly to 88% of control in the CA1 and CA3 regions while lambda binding in the stratum lucidum (CA3) increased to 165% of control. Our results show that opiate receptor subtypes are differentially affected by an ischemic insult. PMID- 2548675 TI - Expression of plasminogen receptors on C6 glioma cells. AB - Plasminogen activators (PAs) play an important role in normal and neoplastic neuromorphogenesis in the central nervous system. Proper function of proteinases such as PA may require focusing of activity on a cellular level. In this study, we demonstrate that highly purified plasminogen binds to receptors on rat C6 glioma cells in culture. Specific binding is reversible and saturable at 4 degrees C. The Kd is 1.95 +/- 0.31 microM and the Bmax is 3.6 x 10(6) molecules/cell. At 37 degrees C, there is no evidence for ligand digestion or internalization. Plasminogen receptors may concentrate potential proteinase near membranes of glia during normal and neoplastic development in the central nervous system. PMID- 2548676 TI - Beta-funaltrexamine disrupts the day-night rhythm of nociception in mice. AB - Determinations were made of the effects of beta-funaltrexamine (beta-FNA), an irreversible mu-opioid receptor antagonist, on the day-night rhythm of nociception in male mice. Peripheral administration of beta-FNA (20 and 40 mg/kg) disrupted the day-night rhythm of foot-licking response to aversive thermal (50 degrees C) stimulation. The peak nocturnal response latency was attenuated and the marked increases and decreases in response latency present at the light-dark and dark-light transitions, respectively, were suppressed. These results suggest that mu opioids are associated with the generation and expression of the day night rhythm of this particular measure of nociception in mice. PMID- 2548677 TI - Opioid inhibition of spontaneously active neurons of the rat arcuate nucleus in vitro. AB - The effects of opioid agonists were determined on single-unit activity recorded from the arcuate nucleus (ARC) in perfused, coronal slices of hypothalamus taken from proestrous rats. The selective, mu-receptor agonist Tyr-D-Ala-Gly-MePhe-Gly ol enkephalin (DAGO) produced a concentration-dependent decrease in the firing rate of 70-78% of the units tested. The concentration of DAGO that induced maximal inhibition of firing was approximately 0.5 microM. This inhibition of firing frequency occurred irrespective of cell location, firing pattern or baseline firing frequency. The effect of DAGO was antagonized by the opioid antagonist naloxone (0.1 microM). The selective, kappa-receptor agonist, trans (+)-3,4 dichloro-N-methyl-[2-(1-pyrrolidinyl) cyclohexyl] benzeneacetamide methane sulfonate (U50,488H) did not decrease the firing rate in cells which did respond to DAGO. Blockade of synaptic activity decreased the level of spontaneous activity but did not prevent the inhibitory action of DAGO. These data support the hypothesis that opioids, through activation of mu-receptors, inhibit neuronal activity in the arcuate nucleus. Furthermore, the opioid inhibition occurs, in part, via a direct postsynaptic action. PMID- 2548678 TI - Beta-carboline interactions at the BZ-GABA receptor chloride-ionophore complex in the rat cerebral cortex. AB - beta-Carboline congeners can act at the benzodiazepine (BZ) recognition site of the BZ-GABA receptor complex to increase GABA-stimulated chloride conductance (agonist effect), inhibit this conductance (inverse agonist effect) or block the actions of agonists and inverse agonists (antagonist effect). In this communication we describe the effects of several beta-carbolines (ZK 93423, ZK 91296, ZK 93426, and DMCM) on GABA-stimulated chloride influx into vesicles prepared from rat cerebral cortex. ZK 93423 produces an approximate 2-fold left shift of the GABA dose-response curve at a concentration of 1.0 microM consistent with its full agonist activity (positive intrinsic efficacy), while the same concentration of ZK 91296 produces over a 1-fold left-shift consistent with its partial agonist activity. At higher concentrations (0.1 mM), ZK 91296 inhibits GABA-stimulated chloride influx which appears to be mediated through a non-BZ receptor mechanism since this effect is not reversed by the BZ antagonist ZK 93426. The augmenting effect of both ZK 93423 and ZK 91296 on GABA-stimulated chloride flux was reduced by the antagonist ZK 93426 in a dose-dependent manner and reached GABA-stimulated control levels at a ZK 93426 concentration of 1.0 microM. Interestingly, there was a further inhibition of the GABA-stimulated chloride influx at higher concentrations of ZK 93426 which is not seen when ZK 93426 is used in the absence of BZ agonists. The inverse agonist activity of DMCM was incompletely blocked by the antagonist ZK 93426. These data show that ZK 93426 can antagonize the effects of the full agonist ZK 93423 and partial agonist ZK 91296 at the BZ receptor. Furthermore, the interaction of the agonists with ZK 93426 results in the appearance of inverse agonist-like activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548679 TI - [The immunodeficiency retrovirus in man and primates]. AB - Human immunodeficiency viruses, HIV-1 and HIV-2, are similar, by genome structure, nucleotide sequences and pathogenicity, to retroviruses existing in various primates, SIVs. The possibility is discussed that the human viruses have their origin in accidental transmission of some monkey viruses to man. Studies of SIVs also suggest that the pathogenic effect not only depends on the virus strain, but also on the host species. PMID- 2548680 TI - [Pharmacokinetics of intradermally-administered drugs]. AB - The dermis, considered as a potential route of administration for drugs, has been widely and empirically used for more than a century. The comparative intravenous and intradermal administration of radiotracers makes it possible to demonstrate that the intradermal route has its own diffusion kinetics and its own biodistribution. PMID- 2548681 TI - [Hepatocellular carcinoma]. PMID- 2548682 TI - Immortalized phenotype and the presence of active oncogenes correlate with the capacity of culture cells to induce reactivation of DNA synthesis in macrophage nuclei in heterokaryons. AB - Several types of culture cells with limited life span (rat embryo fibroblasts, rat chondrocytes and mouse premacrophages) were found to be unable to induce the reactivation of DNA synthesis in the nuclei of non-dividing differentiated cells (mouse peritoneal resident macrophages) in heterokaryons. By contrast, malignant HeLa cells have this ability. In heterokaryons formed by fusion of mouse macrophages with HE239 cells (Syrian hamster fibroblasts transformed with a ts mutant of the SV40 virus), DNA synthesis in macrophage nuclei is reactivated only at the permissive temperature (33 degrees C), at which viral T antigen is stable. Immortalization of rat chondrocytes by transfection with p53 gene enables to induce DNA synthesis in macrophage nuclei upon fusion. All the evidence indicates that the function of immortalizing oncogenes is necessary for the resumption of the DNA synthesis in macrophage nuclei in heterokaryons. PMID- 2548683 TI - [A brief account of the 1988 seminar in Shanghai on viral hepatitis A]. PMID- 2548685 TI - The dental laboratory and you. New system ensures superior resin/metal bond. PMID- 2548686 TI - Osseointegrated implants: five systems. Implants today and the anchor implant system. PMID- 2548684 TI - A dihydropyridine-sensitive calcium channel in rodent osteoblastic cells. AB - Rat osteogenic sarcoma cells (UMR 106-01) and normal rat trabecular bone osteoblasts (ROB) were studied using the whole cell version of the patch clamp technique to determine the existence of calcium (Ca2+) channels. Pipette and bath solutions were designed to separate Ca2+ channel currents from other voltage dependent currents, and Ba2+ was used as the charge carrier. In both UMR 106-01 and ROB cells, a Ba2+ current was measured, which expressed the characteristics of an L-channel, such as activation range, dihydropyridine sensitivity, and little or no inactivation. In some cases, this channel was detectable only with BAY-K-8644 in the bath solution. The dihydropyridine agonist increased the current intensity and shifted the peak inward current to more negative potentials. This study, confirming previous observations, demonstrates the existence of a Ca2+ channel in both transformed and normal osteoblastic cells. PMID- 2548687 TI - Osseointegrated implants: five systems. Integral implant system. PMID- 2548688 TI - Moebius and Poland syndromes: a report of a case. AB - This report describes the dental treatment of a 45-month-old Native American Indian female with Moebius and Poland syndromes, two rare syndromes which can occur simultaneously. PMID- 2548689 TI - Klippel-Trenaunay-Weber syndrome variant. AB - We describe a patient with soft-tissue and bony hypertrophy of all four limbs. The upper limbs were erythematous, with associated finger deformity. The soft tissue irregular hypertrophy and engorgement subsided when the limbs were elevated. Retinal examination showed large, dilated, tortuous veins, with no dye leakage on fluorescein angiography. We feel that this case represents an unusual variant of the Klippel-Trenaunay-Weber syndrome. PMID- 2548690 TI - Effect of albuterol on oxygen uptake and Na+-K+ ATPase activity of jejunal epithelia and liver in chickens. AB - Changes in metabolic characteristics of the liver and jejunal epithelia as a result of feeding the beta-agonist albuterol to chickens for 3 and 6 weeks were studied. Feeding albuterol resulted in a change in weights of the liver and small intestines. Chickens fed albuterol for 6 weeks had greater levels of plasma nonesterified fatty acids than those fed the control diet. Oxygen uptake by liver and jejunal epithelial tissues were 25 and 26% greater for chickens fed the diet with albuterol than for those fed the control diet, respectively. No changes in Na+-K+ ATPase activity of both liver and jejunal epithelia were detected as a result of feeding albuterol. These findings indicate that certain beta-agonists may elevate thermogenesis in chickens as a result of increasing metabolic rates of splanchnic tissues. However, the changes resulting in altering oxygen uptake in liver and jejunal epithelia due to feeding albuterol were not associated with Na+-K+ ATPase activity. PMID- 2548691 TI - L-663,536 (MK-886) (3-[1-(4-chlorobenzyl)-3-t-butyl-thio-5-isopropylindol-2-yl] 2,2 - dimethylpropanoic acid), a novel, orally active leukotriene biosynthesis inhibitor. AB - L-663,536 (3-[1-(4-chlorobenzyl)-3-t-butyl-thio-5-isopropylindol-2-yl]-2, 2 dimethylpropanoic acid) is a potent inhibitor of leukotriene (LT) biosynthesis in intact human polymorphonuclear leukocytes (PMN) (IC50, 2.5 nM). Similarly, L 663,536 inhibited A23187-induced LTB4 formation by rat peripheral blood and elicited PMN. At concentrations where inhibition of leukotriene biosynthesis occurred in human whole blood (1.1 microM), no effect was seen on cyclooxygenase or 12-lipoxygenase, an effect also observed in washed human platelets. The compound had no effect on rat or porcine 5-lipoxygenase indicating that L-663,536 is not a direct 5-lipoxygenase inhibitor. When administered in vivo L-663,536 was a potent inhibitor of antigen-induced dyspnea in inbred rats pretreated with methysergide (ED50, 0.036 mg/kg p.o.) and of Ascaris-induced bronchoconstriction in squirrel monkeys (1 mg/kg p.o.). The compound inhibited leukotriene biosynthesis in vivo in a rat pleurisy model (ED50, 0.2 mg/kg p.o.), an inflamed rat paw model (ED50, 0.8 mg/kg), a model of leukotriene excretion in rat bile following antigen provocation, and a model in the guinea-pig ear where leukotriene synthesis was induced by topical challenge with ionophore A23187 (ED50, 2.5 mg/kg p.o. and 0.6 micrograms topically). The results indicate that L 663,536 is a potent inhibitor of leukotriene biosynthesis both in vitro and in vivo indicating that the compound is suitable for studying the role of leukotrienes in a variety of pathological situations. PMID- 2548692 TI - Superoxide radical production in plasma membrane samples from regressing rat corpora lutea. AB - Plasma membrane samples prepared from regressing rat corpora lutea were examined for production of the superoxide radical. A procedure was developed to purify membrane samples that were enriched approximately 15-fold with the plasma membrane marker enzyme, and superoxide radical levels were determined using electron spin resonance to measure Tiron semiquinone. During prostaglandin F2 alpha-induced and spontaneous regression, there was a significant increase in formation of superoxide radical that was not observed in plasma membrane samples from nonregressing corpora lutea. Plasma membrane incubation experiments indicated that the increase in production was temperature sensitive and reduced with inhibitors of phospholipase A2 and cyclooxygenase. Addition of superoxide dismutase or vitamin E abolished superoxide radical formation in vitro. Following the rise in superoxide radical levels during regression, there was also a significant decrease in the activity of the plasma membrane enzyme, Na+-K+ ATPase. These results indicate that the production of superoxide radical increases in plasma membrane samples prepared from regressing rat corpora lutea and that this increase is mainly due to the products of phospholipase A2 and cyclooxygenase activity. PMID- 2548693 TI - Ca2+ channels in chick neural retina cells characterized by 1,4-dihydropyridine antagonists and activators. AB - The voltage-sensitive calcium channel in cultured chick neural retina cells was characterized by the actions of the enantiomers of Bay K 8644 and 202-791 and other 1,4-dihydropyridines. These cells showed time- and voltage-dependent Ca2+ uptake that was stimulated by K+ depolarization and blocked by the inorganic calcium channel blockers Cd2+ and Co2+. A small fraction only (15% maximum) of the uptake was inactivated by predepolarization of the cells with 80 mM K+. Ca2+ uptake was sensitive to the 1,4-dihydropyridine calcium channel antagonists and activators. (S)-Bay K 8644 and (S)-202-791 stimulated the Ca2+ uptake, and (R) Bay K 8644 and (R)-202-791 as well as nitrendipine and PN 200-110 inhibited Ca2+ uptake stimulated by K+ depolarization or channel activators. The K+ depolarization-stimulated uptake was inhibited by 90%, but the activator stimulated uptake was completely blocked by the 1,4-dihydropyridine antagonists. The potencies of these agents as inhibitors of Ca2+ uptake were significantly lower than the binding affinities in membrane preparations from the same cells or their binding and pharmacologic affinities in vascular smooth muscle. K+ depolarization or (S)-Bay K 8644 induced 45Ca2+ uptake was not observed in a glial cell culture. [3H]Nitrendipine and [3H]PN 200-110 bound to membrane preparations of the cells consistent with the presence of a single type of high affinity binding site.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548694 TI - Relationship between oxidative phosphorylation and adenine nucleotide translocase activity of two populations of cardiac mitochondria and mechanical recovery of ischemic hearts following reperfusion. AB - The possible relationship of the atractyloside-sensitive adenine nucleotide translocase activity, oxidative phosphorylation, and the recovery of ventricular contractility following reperfusion of the ischemic isolated rat heart was studied. Five minutes of total global ischemia without reperfusion produced a significant depression in adenine nucleotide translocase in subsarcolemmal mitochondria (SLM), whereas a minimum of 10 min ischemia was required to observe a significant depression in interfibrillar mitochondria (IFM). Increasing durations of ischemia resulted in a progressively larger depression in translocase activity, with a maximum depression of approximately 75% seen in both populations following 20 min ischemia. In contrast, oxidative phosphorylation was totally unaffected in either mitochondrial population following up to 20 min of ischemia. We assessed whether translocase activity or oxidative phosphorylation were related to contractile recovery in hearts reperfused following various durations of ischemia. In SLM, translocase activity was further depressed following reperfusion compared with pre-reperfusion ischemic values, whereas with IFM only reperfusion following 5 min ischemia produced a further depression in translocase values. Oxidative phosphorylation rates of SLM and IFM were significantly depressed following reperfusion of ischemic hearts, although SLM exhibited a generally higher sensitivity in this regard. In reperfused hearts, an overall significant relationship was found between oxidative phosphorylation rate and adenine translocase activity as well as between translocase activity and post reperfusion contractile recovery. These data show that ischemia can produce a significant depression in translocase activity in the absence of any change in oxidative phosphorylation. The results also suggest that the depression in mitochondrial ADP/ATP translocase and subsequent inhibition of oxidative phosphorylation in the reperfused heart may represent one of the important contributory mechanisms involved in cardiac failure and injury during acute ischemia and reperfusion. PMID- 2548695 TI - The mechanism of the rate-dependent changes of the conducted action potential in rabbit ventricle. AB - Blockers of the transient outward current (4-aminopyridine) and the Ca current (Co2+) as well as injection of polarizing current during the plateau were used to assess the role of these current systems as determinants of action potential duration at different pacing rates. Papillary muscles and ventricular trabecula were superfused with oxygenated Krebs solution at 33 degrees C and driven at a basic rate of 1 Hz. The effects of varying the frequency of stimulation between 0.1 and 4 Hz on action potential parameters were determined under control conditions and during exposure to 2 mM 4-aminopyridine, 1-3 mM CoCl2, or a mixture of 4-aminopyridine and CoCl2. The control relationship between action potential duration and pacing rate showed a maximum between 1 and 2 Hz. Under 4 aminopyridine, the plateau height and the action potential duration increased. The rate-dependent shortening of the action potential at frequencies below 1 Hz was reduced or abolished, and enhanced shortening was observed at rates above 1 Hz. Exposure to Co2+ reduced the action potential shortening at rates higher than 1 Hz. Both blockers, 4-aminopyridine and Co2+ were necessary to eliminate the rate-dependent changes of the action potential duration. Our results indicated that both the transient outward current and the inward calcium current determine the plateau height and duration for frequencies less than or equal to 2 Hz, whereas at higher rates, the Ca current plays a dominant role. PMID- 2548696 TI - Accumulation of incomplete metabolic side products of lipid A in Salmonella typhimurium during inhibition of 3-deoxy-D-manno-octulosonate incorporation by a new class of antibacterial agents. AB - A new class of antibacterial agents for Gram-negative bacteria, rationally designed to inhibit the incorporation of 3-deoxy-D-manno-octulosonate into lipopolysaccharide (LPS), was recently reported. In Salmonella typhimurium, where the lipid A species are well characterised, it was previously demonstrated that the addition of a compound which inhibits the enzyme 3-deoxy-manno-octulosonate cytidylytransferase (CMP-KDO synthetase; EC 2.7.7.38) leads to rapid accumulation of lipid A derivatives. The major lipid A species, IVA (O-(2-amino-2-deoxy-beta-D glucopyranosyl)-(1-6)-2-amino-2-deoxy-alpha-D - glucose, acylated at positions 2, 3, 2', 3' with beta-hydroxymyristoyl groups and bearing phosphates at positions 1 and 4'), was shown to be converted mainly to LPS by pulse-chase experiments in the absence of inhibitor. Labelled precursor (IVA) was also chased to other more polar lipid A derivatives. During chase in the presence of inhibitor, there was no conversion to LPS, while the major lipid A species was converted to the same polar lipid A derivatives as in chase without inhibitor. Our data indicate that despite the accumulation of several species of lipid A derivatives during inhibition of LPS synthesis, only IVA is destined for synthesis of mature LPS when LPS synthesis resumes. The more polar lipid A derivatives would thus represent aberrant side reaction products which occur when the pathway is inhibited. PMID- 2548697 TI - Neuropathology of heart transplantation. AB - The neuropathology of 18 cardiac transplant recipients was reviewed with the clinical findings. Pathological changes were noted in the central nervous system (CNS) in 94% of the patients, the most frequent being cerebral vascular in origin (72%). Eight patients (44%) had multiple cerebral infarcts and morphologically, a large number of these antedated the transplantation. In addition 4 patients had acute focal ischemic changes which occurred after transplantation. Intracranial hemorrhage was noted in 5 patients (28%), including one case of fatal intracerebral hemorrhage following an acute hypertensive episode after the transplantation. While systemic infection was common (10 patients), there were only 5 cases of intracranial infection; including 3 cases of cytomegalovirus infection, one of candidiasis and one of aspergillosis. Post-transplant seizures, occurring in a third of the patients, were related to a variety of causative factors such as sepsis, intracranial hemorrhage, cerebral ischemia, metabolic encephalopathy and cyclosporin neurotoxicity. Of note in this series was the absence of CNS lymphoma or other systemic lymphoproliferative disorder. PMID- 2548698 TI - Mental hospital depopulation in Canada: patient perspectives. AB - This paper reviews briefly the history of mental health depopulation in Canada over the past 30 years. The term "deinstitutionalization" is often used but is unsatisfactory. Using an exploratory, qualitative, methodological approach, data were collected on the problems encountered by a disproportionate, stratified random sample of 139 formerly institutionalized patients living in various geographical locales in Eastern Canada. Adopting a symbolic interactionist theoretical approach, this study, in an effort to fill a neglect in the literature, attempted to discover what the everyday world(s) of Canadian ex mental patients was really like. Problems encountered related to stigma, poor housing, lack of back living skills, poverty, unemployment and aftercare. Quotations from patients are provided to illustrate such themes. The findings are discussed. PMID- 2548699 TI - The establishment of Epstein-Barr virus nuclear antigen-positive (SP-50B) and Epstein-Barr virus nuclear antigen-negative (SP-53) cell lines with t(11;14)(q13;q32) chromosome abnormality from an intermediate lymphocytic lymphoma. AB - Two lymphoma cell lines, SP-50B and SP-53, were established from peripheral blood of a 58-year-old woman with leukemic conversion of intermediate lymphocytic lymphoma. These cell lines grew in suspension with or without forming clumps of cells. SP-50B was morphologically similar to the common Epstein-Barr (EB) virus transformed lymphoblastoid cell lines and was positive for EB virus nuclear antigen (EBNA), whereas SP-53 closely resembled the patient's lymphoma cells and was negative for EBNA. Both cell lines expressed the same phenotypic markers as original lymphoma cells (CpIg+, SmIg+, OKIa1+, Leu12+) and possessed t(11;14)(q13;q32) chromosome translocation. These results indicate that although morphologically different, SP-50B and SP-53 were both derived from patient's lymphoma cells. The long-term cultivation of EBNA-positive and EBNA-negative B cell lymphoma lines from a single donor has not been previously reported. These cell lines would provide useful tools for studying the oncogenic role of EB virus and bcl-1 oncogene that is located on chromosome 11q13. PMID- 2548700 TI - Image and flow DNA cytometry of small cell carcinoma of the lung. AB - Both image and flow DNA cytometry were performed in isolated nuclei from paraffin embedded tumor tissue of patients with small cell carcinoma of the lung (SCCL). In 14 patients tissue was obtained by surgery from the primary tumor. From 14 patients tissue was taken by autopsy. From two patients tissue obtained by both surgery and later autopsy were available. From the autopsy patients tissue was taken only from the primary tumor (n = 6), from a metastasis (n = 1) and from the primary tumor and distant metastases (n = 7). Twelve of the tumors obtained by surgery were diploid, and two multiploid (two stem lines present). This was found both with image and flow cytometry. The group of patients could clearly be subdivided in short survivors (less than 9 months, n = 6) and long survivors (greater than 16 months, n = 8); since in both groups one multiploid and the remainder diploid cases were present, ploidy did not seem to be a good prognosticator for survival. In most (n = 26) of the tissues measured from the autopsy patients, again, a good correlation between image and flow DNA cytometry was obtained, the histograms being either (near) diploid or multiploid. In six cases, however, flow cytometry showed multiploidy whereas image showed aneuploidy (one single peak clearly deviating from diploidy). This discrepancy is caused because normal diploid (nonneoplastic) cells in the preparations could not be discarded from the flow cytometry measurements. Using the image cytometry data of the primary tumors, five diploid, three aneuploid, and four multiploid tumors were found. In five of the seven patients of whom tissue was obtained from the primary tumor and multiple metastases, differences between the histograms were found, mostly showing two malignant cell populations in one tissue and only one of them in another. Of one of the two patients of whom tissue was obtained by surgery and later autopsy, a change in histogram pattern was observed. It is concluded that although there is a high similarity between image and flow DNA cytometry, for an optimal interpretation of the histogram pattern, image measurements are more reliable. Ploidy determination does not seem to be of use in prediction of survival, and care should be taken in interpreting DNA histograms of metastases in SCCL patients because of the variability in histogram pattern. PMID- 2548701 TI - Pulmonary sclerosing hemangioma of the lung. A type II pneumocytoma by immunohistochemical and immunoelectron microscopic studies. AB - Three cases of pulmonary sclerosing hemangioma were studied by immunohistochemical and immunoelectron microscopic methods using a panel of antibodies. Six cases of adenocarcinoma of the lung, three cases of normal mesothelium, and three cases of mesothelioma were used as controls. The cytoplasm of some of the sclerosing hemangioma tumor cells was positive for the anti-lung surfactant apoprotein monoclonal antibody (PE-10). These cells were the pale cells of the solid areas, the cells covering the papillary projections, and the cells lining the cleft-like spaces. These cells also were positive for conventional epithelial cell markers. Some cells also were positive for vimentin. Electron microscopic study showed that the predominant cell was a poorly differentiated pneumocyte. Immunoelectron microscopic study also demonstrated that PE-10 existed in the rough endoplasmic reticulum of some of the cells in the solid areas, in the same way as normal type II pneumocytes. We concluded that the sclerosing hemangioma is an epithelial tumor with differentiation towards type II pneumocytes. PMID- 2548702 TI - A case of biliary cystadenocarcinoma with morphologic and histochemical features of hepatocytes. AB - A case of biliary cystadenocarcinoma is presented in which the lesion seemed to partially differentiate into hepatocellular carcinoma. This patient had a cystic tumor in the left lobe of the liver, the interior being filled with papillary tumors. Histologic examination disclosed papillary growth of columnar tumor cells, together with tumor cells akin to hepatocytes (they had a distinct cell boundary, nuclear membrane, and nucleolus) showing a cobblestone appearance in some areas. Electron microscopic examination of this area showed bile canalicular structures. Histochemically, the tissue stained positively with the Luna-Ishak canalicular (LIC) technique; periodic acid-Schiff (PAS) staining demonstrated coexisting glycogen granulo-positive cells and mucus-producing cells. Thus, in this biliary cystadenocarcinoma, some tumor cells demonstrated the morphologic and histochemical features of hepatocytes. PMID- 2548703 TI - Intraductal papillary neoplasms of the pancreas. A clinicopathologic study of six patients. AB - A clinicopathologic study was conducted on six patients with intraductal papillary neoplasms of the pancreas. The patients were of both sexes and their ages ranged from 64 to 79 years. Three patients had a long history of symptoms mimicking chronic pancreatitis. The tumors involved the main pancreatic duct in the head-body region either diffusely or focally. Histologic examination showed papillary proliferations of well-differentiated, mucus-secreting cells that occasionally stained for carcinoembryonic antigen (CEA) and carbohydrate antigen (CA 19-9). The proliferations filled the main pancreatic duct, and extended into smaller ducts in some cases. In three patients, the lesions contained foci of pronounced to severe cellular atypia and carcinoma in situ. None of our series or any similar cases reported in the literature has shown invasion into peripancreatic tissue, metastasis, or tumor recurrence after pancreatectomy. Because of their favorable prognosis, intraductal papillary neoplasms should be considered low-grade malignancies that must not be confused with the common ductal adenocarcinoma. PMID- 2548704 TI - Small cell carcinoma of the urinary bladder. A clinicopathologic, morphometric, immunohistochemical, and ultrastructural study of 18 cases. AB - A multiinstitutional review of 3778 patients with a primary malignancy of the urinary bladder revealed 18 cases (0.48%) of small cell carcinoma which were histologically and morphometrically identical to pulmonary small cell undifferentiated carcinoma. Age, sex, and symptoms at first presentation were comparable to that known in transitional cell carcinoma. Sixteen patients (89%) developed metastatic disease, with most frequent involvement of regional lymph nodes, liver, skeleton, and abdominal cavity. The unfavorable clinical outcome was worse as compared with that reported in advanced stage poorly differentiated transitional cell carcinoma, and was similar to the rapidly fatal outcome of pulmonary small cell undifferentiated carcinoma. Fourteen patients (78%) died by tumor at a mean follow-up period of 9.4 months, and only one patient was free of recurrent disease more than 5 years after cystectomy. This apparent aggressive tumor behavior was independent of the presence of neuroendocrine differentiation characteristics at immunohistochemical (13 cases, 72%) or electron microscopic study (eight cases, 44%). The prolonged survival periods (15-38 months) of the five patients who received combination chemotherapy suggested that, just as in small cell lung carcinoma, chemotherapy may be profitable. A unified concept of histogenesis of bladder cancer with a common origin from a multipotent mucosal stem cell is proposed. PMID- 2548705 TI - Translocation 10;17 in clear cell sarcoma of the kidney. A first report. AB - This is the first report of a t(10;17) as the unique cytogenetic finding in one case of a rare childhood tumor, clear cell sarcoma of the kidney (CCSK). This observation is discussed in relation to the cytogenetics of Wilms' tumors, of which CCSK is a variant. PMID- 2548706 TI - Hyperdiploidy including trisomy 8 in a cystic partially differentiated nephroblastoma. AB - Cystic partially differentiated nephroblastoma (CPDN), a rare, cystic, renal lesion of childhood, has not been previously karyotyped. It is distinguished histologically from multilocular renal cyst by the presence of blastemal cells, and from Wilms' tumor by lack of expansile, solid growth and by indolent clinical behavior. In the present case, ten of 20 analyzed cells from a 3-week culture obtained from the tumor had a clonal, hyperdiploid karyotype. The modal chromosome number was 51, with chromosomes 8, 12, 17, 19, and 20 usually being present in three copies. Trisomy 8 was present in every hyperdiploid cell examined. A normal 46,XY constitutional karyotype was also observed. In degree and significance, the hyperdiploidy of CPDN is thus distinct from that reported in the prognostically unfavorable, anaplastic Wilms' tumor, where the DNA index is typically near-tetraploid. Trisomy 8, as a constitutional mosaicism, has been previously reported in children with bilateral CPDN and/or undifferentiated sarcomas, although none of their tumors were karyotyped. The present findings support a neoplastic nature for CPDN, while emphasizing its pathogenetic distinctiveness from Wilms' tumor, and provide further evidence for significance of trisomy 8 in the pathobiology of this tumor. PMID- 2548707 TI - Protein-linked DNA strand breaks induced in mammalian cells by camptothecin, an inhibitor of topoisomerase I. AB - Camptothecin was recently identified as an inhibitor of mammalian topoisomerase I. Similar to inhibitors of topoisomerase II, camptothecin produces DNA single strand breaks (SSB) and DNA-protein cross-links (DPC) in mammalian cells. However, their one-to-one association, expected for trapped topoisomerase complexes, has not previously been demonstrated. We have studied camptothecin induced SSB and DPC in Chinese hamster DC3F cells and their isolated nuclei, using the DNA alkaline elution technique. It was found that the SSB and DPC frequencies detected following camptothecin treatment depend upon the conditions used for lysis. When lysis was with sodium dodecyl sulfate, the observed frequencies of SSB and DPC were 2- to 3-fold greater than when sodium dodecyl sarkosinate (Sarkosyl) was used. In either case, the SSB:DPC ratio was close to 1. All of the camptothecin-induced SSB were protein linked, as indicated by the absence of DNA elution under nondeproteinizing conditions. DNA cleavage assays with purified topoisomerase I also indicated that the weaker Sarkosyl detergent fails to trap all of the enzyme-DNA complexes. In contrast, lysis conditions had little effect on levels of SSB or DPC produced by 4'-(9-acridinylamino) methanesulfon-m-anisidide, suggesting that trapping of topoisomerase II complexes occurs equally well with either detergent. In experiments using isolated nuclei, it was found that the camptothecin-induced SSB, in contrast to trapped topoisomerase II complexes, can form and reverse within minutes at 4 degrees C. The activity of camptothecin at low temperature was also seen with purified topoisomerase I. These results support the hypothesis that the SSB and DPC induced by camptothecin in mammalian cells are due to an action on topoisomerase I. PMID- 2548708 TI - Immunological and pharmacological removal of small cell lung cancer cells from bone marrow autografts. AB - Autologous bone marrow transplantation is used in small cell lung cancer (SCLC) to reverse the hematological toxicity induced by high dose therapy even though the presence of cancerous cells in the graft is potentially dangerous by reinfusion of the disease along with the hematopoietic stem cells. The present studies were undertaken to examine the effectiveness of anti-SCLC rat monoclonal antibodies LCA1 and LC66 plus human complement combined with a derivative of cyclophosphamide (Asta-Z 7557) for the elimination of cancerous clonogenic cells from the graft. In a series of assays conducted with three SCLC cell lines, used alone or mixed with normal bone marrow cells, the addition of Asta-Z 7557 to two cycles of treatment with monoclonal antibodies plus complement results in a 4- to 5-logarithmic reduction of the clonogenic SCLC cells detectable by limiting dilution analysis. This was superior to either treatment used alone. When normal bone marrow was submitted to the same treatment, a median (range) of 44% (15-77%) of the colony-forming unit, granulocyte-macrophage was recovered. These results suggest that the association of immunological (LCA1 and LC66 plus human complement) and pharmacological (Asta-Z 7557) removal methods is effective for purging metastatic clonogenic cells from bone marrow of SCLC patients and could be considered before autologous bone marrow transplantation. PMID- 2548709 TI - Effects of irradiation on the release of growth factors from cultured bovine, porcine, and human endothelial cells. AB - The effects of radiation on the release of mitogenic factors into the media of cultured endothelial cells of bovine, porcine, and human origins were studied. Although unirradiated controls revealed a significant background activity, single doses of irradiation (20-60 Gy) resulted in a dose-related increased release of growth factor activity, measured by the mitogenic effects of the conditioned media on both 3T3 mouse fibroblasts and unirradiated endothelial cells serving as target cells. Receptor binding competition assays for the platelet-derived growth factor receptor revealed that 12-28% of the total mitogenic activity was due to platelet-derived growth factor-like mitogens. Mitogenic assays using endothelial cells and specific antibody mediated inhibition assays suggested that another component of the mitogenic activity was due to a fibroblast growth factor-like factor. Although radiation resulted in a significant increase in cell death, the enhanced growth factor activities did not appear to result from cell lysis related leakage of intracellular stores of growth factor. Instead, our data suggest that the growth factors were synthesized de novo and secreted at elevated levels by the cells which maintained postradiation a high level of metabolic activity. Time course studies demonstrated that the growth factors accumulation in the conditioned media started within the first 24 h after radiation and reached a plateau within 72 h after treatment. Radiation-induced release of endothelial cell-derived growth factors may be involved in the pathogenesis of both early vascular damage and the late fibrosis which represents a prominent feature of late radiation damage in normal tissues. PMID- 2548710 TI - Arrest of replication forks by drug-stabilized topoisomerase I-DNA cleavable complexes as a mechanism of cell killing by camptothecin. AB - Camptothecin, which induces an unusual type of DNA damage by trapping cellular topoisomerase I on chromosomal DNA in the form of drug-enzyme-DNA cleavable complexes, inhibits DNA synthesis and specifically kills S-phase cells. Cotreatment of L1210 cells with aphidicolin, which is an inhibitor of replicative DNA polymerases, completely abolished camptothecin cytotoxicity, suggesting the involvement of DNA replication in camptothecin cytotoxicity. In order to study the role of DNA replication in drug action, a cell-free SV40 DNA replication system was used in the present study. Camptothecin inhibited SV40 DNA replication in this cell-free system only in the presence of topoisomerase I. Addition of excess purified calf thymus DNA topoisomerase I to this extract system in the presence of camptothecin resulted in severe inhibition of SV40 DNA replication and the accumulation of linearized replication products, which contained covalently bound DNA topoisomerase I. We propose that the collision between moving replication forks and camptothecin-stabilized topoisomerase I-DNA cleavable complexes results in fork arrest and possibly fork breakage, which are lethal to proliferating cells. PMID- 2548711 TI - Antigenic profile of tumor progression stages in human melanocytic nevi and melanomas. AB - Sixteen monoclonal antibodies that were obtained after immunization of BALB/c mice with intact melanoma cells or extracts of melanoma cells were tested for reactivity with normal and malignant melanocytic cells in situ, using an immunoperoxidase technique on frozen tissue sections. Sections representing six histopathologically defined stages of tumor progression, ranging from normal melanocytes to highly malignant metastatic lesions, were used. Thirteen monoclonal antibodies (MAbs) did not stain normal melanocytes in situ, whereas three MAbs weakly stained between 1 and 12.5% of melanocytes in 6-22% of the skin sections examined. MAb B 73.1, which was produced by immunization of mice with human natural killer cells and which binds to the Fc receptor of natural killer cells and granulocytes, reacted exclusively with malignant cells that represent the last two stages of tumor progression, vertical growth phase (VGP) primary melanoma and metastatic melanoma. All other antibodies showed variable reactivity with benign proliferative lesions or radial growth phase (RGP), an early stage of primary melanoma. Staining by MAbs that were reactive with gangliosides, unknown antigens, receptors, and two proteins (120/94 kDa protein and 250 kDa glycoprotein) showed a gradual increase in subsequent stages of tumor progression. Two steps in tumor progression were characterized by significant quantitative changes in the expression of antigens detected by the MAbs used in this study. First, mature nevus cells showed significantly higher reactivity with a panel of six MAbs, when compared to normal melanocytes. Second, a separate panel of six MAbs discriminated between RGP and VGP primary melanoma cells. No significant differences in antigen expression were found between dysplastic nevus cells and RGP melanoma, except that some antigens (nerve growth factor receptor and GD2/GD3 gangliosides) appear to be expressed at lower levels in RGP lesions, nor did VGP primary and metastatic melanomas show significant differences in antigen expression. These results suggest that (a) tumor progression of melanocytic cells in vivo is accompanied by significant quantitative differences in the expression of antigens, (b) some of the antigens examined here are associated with biologically aggressive malignant lesions but not normal or premalignant melanocytic cells, and (c) RGP primary melanoma cells are antigenically more similar to nevus cells than to VGP primary melanoma cells. PMID- 2548712 TI - Inhibitory effects of the tyrosine kinase inhibitor genistein on mammalian DNA topoisomerase II. AB - Tyrosine phosphorylation plays a crucial role in cell proliferation and cell transformation which suggests that tyrosine kinase-specific inhibitors might be used as anticancer agents. When the cytotoxic effect of the potent tyrosine kinase inhibitor genistein on various cell lines was studied, we observed that 9 hydroxyellipticine-resistant Chinese hamster lung cells (DC-3F/9-OH-E) were markedly more resistant to genistein than the parental cell line (DC-3F). The DC 3F/9-OH-E cells have been shown to have an altered DNA topoisomerase II activity. We therefore examined the effects of genistein on DNA topoisomerase II-related activities of nuclear extracts from DC-3F cells as well as on purified DNA topoisomerase II from calf thymus. Our results show that genistein (a) inhibits the decatenation activity of DNA topoisomerase II and (b) stimulates DNA topoisomerase II-mediated double strand breaks in pBR322 DNA on sites different from those of 4'-(9-acridinylamino)methanesulfon-m-anisidide, etoposide, and 2 methyl-9-hydroxyellipticinium. Structure-activity studies with six chemically related compounds show that only genistein has an effect on the cleavage activity of DNA topoisomerase II in the concentration range studied. Finally, genistein treatment of DC-3F cells results in the occurrence of protein-linked DNA strand breaks as shown by DNA filter elution. Viscometric (lengthening) studies demonstrate that genistein is not a DNA intercalator. Genistein is therefore an interesting compound because it induces cleavable complexes without intercalation. Taken together, our results show that genistein is an inhibitor of both protein tyrosine kinases and mammalian DNA topoisomerase II. This could be accounted for by the sharing of a common structure sequence between the two proteins at the ATP binding site. PMID- 2548713 TI - Lung cancer-associated monoclonal antibody 15 that recognizes cell growth-related membrane antigens gp85/45 and its growth-inhibitory effect on human lung cancer cells in vitro. AB - In order to find lung cancer-specific markers, monoclonal antibody 15 (MAb15) was produced against a variant-type cell of small cell lung carcinoma. Its gp85/45 antigens were demonstrated in 70% of lung cancers, and particularly in the proliferating zone of cancer cell nests, but they are scarcely detected in noncancerous tissues. Immunoelectron microscopy revealed that gp85/45 antigens were expressed alternatively on the cell membrane of living cancer cells according to their biological states. MAb15 added to the culture medium inhibited the proliferation of lung cancer cells, depending on its concentration, but cell death rate did not increase. The growth inhibition by MAb15 was reevaluated by a colonogenic assay. On DNA histogram, MAb15 decreased the number of DNA synthesizing cells in the S phase with an elevation of the G1 peak, indicating a G1-S boundary block in the cell cycle. gp85/45 detected by this lung cancer associated monoclonal antibody could be a functional membrane unit, such as a growth factor receptor, which is related to the cell proliferation of lung cancer. The growth inhibition by MAb15 may be caused by the blocking of a growth factor receptor which is specific to lung cancer. PMID- 2548714 TI - Establishment and characterization of a human ovarian neoplastic cell line, DO-s. AB - A permanent human neoplastic cell line, DO-s, was established from ascites of a patient with a well-differentiated mucinous cyst-adenocarcinoma of the ovary. This cell line grew as vermiform, floating colonies of epithelial cells in culture. The karyotype of DO-s was of a human female; the chromosome number ranged from 54 to 66 with several abnormalities, mainly trisomy. Epithelial-like character was confirmed by transmission electron microscopy and by the presence of cytokeratin. Inoculation of DO-s cells i.p. or s.c. in athymic nude mice resulted in, respectively, ascites and xenografts. Light and electron microscopical analysis of cultured cells and xenografts demonstrated that the cell line was derived of a mucinous adenocarcinoma biopsy. Tumor-associated antigens, cancer antigen 125 (CA 125), human milk fat globulin, and human placental alkaline phosphatase were expressed by cells in culture and in xenografts. Modulation of the antigens, CA 125 and human milk fat globulin, occurred in DO-s cells growing in athymic mice. Biochemical, immunohistochemical, and histochemical analysis showed that more than 50% of the alkaline phosphatase isoenzymes present in DO-s cells had the characteristics of human placental alkaline phosphatase and placental alkaline phosphatase-like alkaline phosphatase (AP), but fractions of intestinal AP and nonspecific AP (bone-liver-kidney) were also present. The expression of AP isoenzymes could be induced by an enhancement of the serum supplement in the culture media, and by dexamethasone, sodium butyrate, and bromodeoxyuridine. This line will be a valuable tool in studying the therapeutic effects of antibodies to tumor-associated antigens or other agents for ovarian cancer. PMID- 2548716 TI - Loss of calcitonin receptors: a genetically transmitted defect in rats with high incidence of C-cell tumors. AB - C-cell tumors (medullary thyroid carcinoma) occur in humans and several other mammalian species. This tumor develops spontaneously with a high incidence (50%) in old Wag/Rij (Wistar-derived strain) rats. We have recently shown that calcitonin binding sites, which are present in the Wistar rats, are lost from renal medulla of the Wag/Rij rats before they reach the age of 1 month. In the present work, we investigated the distribution of calcitonin binding sites in the kidneys of first and second generation hybrids of Wistar x Wag/Rij rats. The absence of calcitonin binding sites from the renal medullas of 25% of F2 hybrids indicates that the deficiency is inherited in a Mendelian fashion and opens the way to establishing inbred strains lacking renal medullary calcitonin binding sites. PMID- 2548715 TI - Altered function of protein kinase C and cyclic adenosine monophosphate-dependent protein kinase in a cell line derived from a mouse lung papillary tumor. AB - Two serine/threonine protein kinases were compared in C10, a clone from the nontumorigenic NAL IA cell line derived from normal mouse lung epithelium, and PCC4, a cell line derived from a mouse lung adenoma. C10 cells are contact inhibited, whereas PCC4 cells are not. Upon treatment with the phorbol ester, 12 O-tetradecanoylphorbol-13-acetate (TPA), the normally flattened C10 cells round up, while the normally bipolar, rounded PCC4 cells flatten out. Three proteins of 14,000, 20,000 and 116,000 molecular weight were phosphorylated in TPA-treated particulate fractions but not in untreated particulate fractions of PCC4 cells. In contrast, TPA caused a generalized increase in the phosphorylation of most membrane proteins in C10 cells. Cytosolic protein kinase C (PKC) specific activity was lower in PCC4 cells than in C10 cells, but particulate PKC activity was similar in the two cell lines. Both measurements of PKC activity and immunoblotting assays using anti-PKC antisera showed increased particulate PKC in TPA-treated C10 cells resulting from a quantitative translocation of PKC molecules from cytoplasm to plasma membrane. This PKC response to TPA was attenuated in PCC4 cells. While PCC4 particulate PKC activity was substantially increased after TPA treatment, PKC activity decreased only slightly in cytosolic fractions of TPA-treated PCC4 cells. Immunoblots of TPA-treated PCC4 cells showed a decline in cytosolic PKC content and increased particulate PKC concentration, but these changes were not of the same magnitude as the activity changes. This may represent an unmasking of latent PKC activity since particulate PKC activity in TPA-treated PCC4 cells was inhibited by staurosporine, a specific inhibitor of PKC when used at nanomolar concentrations. In addition, PCC4 cells had less mRNA coding for the R1 regulatory subunit of cyclic AMP (cAMP)-dependent protein kinase (PKA) than C10 cells, as determined by Northern blotting using an R1 alpha cDNA probe. Consistent with this result, photolabeling with 8-azido-[32P]cAMP, a photoaffinity analog of cAMP, revealed that R1 from PCC4 cells incorporated less analogue than R1 from C10 cells. PKA-specific activity also was lower in PCC4 cells than in C10 cells. Thus, deficiencies in protein kinases which mediate the effects of diacylglycerol and cAMP second messengers were observed in neoplastic lung cells. This may dampen the responsiveness of PCC4 cells to extracellular signals that regulate cell growth and cell-cell interactions. PMID- 2548717 TI - Antitumor activity and nucleic acid binding properties of dercitin, a new acridine alkaloid isolated from a marine Dercitus species sponge. AB - A new cytotoxic acridine alkaloid that exhibited antitumor activity in vivo was isolated from a marine Dercitus species sponge collected at a depth of 160 m in the Bahamas. This violet alkaloid, designated dercitin, inhibited the proliferation of cultured murine and human leukemia, lung, and colon tumor cells at nM concentrations (IC50 values of 63-150 nM) and prolonged the life of mice bearing ascitic P388 tumors (%T/C = 170, 5 mg/kg, i.p., QD1-9). Dercitin was also active against i.p. B16 melanoma and modestly inhibited the growth of s.c. Lewis lung carcinoma on the same schedule. DNA blocked the antiproliferative effects of the agent in culture, and incorporation studies indicated that dercitin disrupted DNA and RNA synthesis with less effects on protein synthesis, similar to the effects of known DNA intercalators. After 1-h exposure to 400 nM dercitin, the rates of incorporation of [3H]uridine, [3H]thymidine, and [3H]leucine by cultured P388 cells were inhibited 83, 61, and 23%, respectively. Equilibrium dialysis indicated that dercitin bound calf thymus DNA with an affinity of 3.1 microM and maximal binding of 0.20 mol dercitin/mol base pair. Binding involved intercalation as evidenced by ability to relax supercoiled phi X174 DNA (half maximal concentration for dercitin relaxation was 36 nM). The effects of dercitin on DNA mobility were reversible, and complete relaxation of DNA with topoisomerase I in the presence of dercitin followed by phenol extraction resulted in the appearance of supercoiled DNA. Dercitin, at microM concentrations, had a small effect in the K+-sodium dodecyl sulfate assay using cultured P388 cells, suggesting minimal inhibition of topoisomerase activity. But, dercitin completely inhibited DNA polymerase I/DNase nick translation of DNA at 1 microM. Relaxation of DNA at a given concentration was greater than inhibition of nick translation suggesting that the effects of dercitin on enzyme activity were secondary to changes in DNA conformation. Results indicate that dercitin is a new marine natural product that probably exerts its biological effects through intercalation into nucleic acids. PMID- 2548718 TI - Antiproliferative and differentiative effect of granulocyte-macrophage colony stimulating factor on a variant human small cell lung cancer cell line. AB - A variant clone was adopted during passages of a small cell lung cancer cell line, GKT3-1.3. The variant clone exhibited distinct characteristics with alterations in morphology, positive staining with nonspecific esterase stain, and an increase in surface specific markers OKM5, HLA-DR, Mo1, and My7, usually found on monocytes or their precursors. However, it exerted a very rapid proliferation just like immature cells. This new clone, GKT3-1.3V, was shown to have specific binding capacity to granulocyte-macrophage colony-stimulating factor (GM-CSF), with a number of binding sites comparable to that of myelomonocytes or monocytic cell lines. Thus its proliferation was inhibited by GM-CSF in clonogenic assay and suspension culture. Increase in the percentage of cells with surface marker Mo1 by the addition of GM-CSF suggested its differentiative effect. Cell cycle analysis showed that the antiproliferative effect of GM-CSF was due to a block in G0 or G1. The antiproliferative effect of GM-CSF was abolished by the addition of anti-GM-CSF antibody. PMID- 2548719 TI - Heterogeneity of O6-alkylguanine-DNA alkyltransferase activity in peripheral blood lymphocytes: relationship between this activity in lymphocytes and in lymphoblastoid lines from normal controls and from patients with Hodgkin's disease or non-Hodgkin's lymphoma. AB - We determined O6-alkylguanine-DNA alkyltransferase (AGT) activity in the peripheral blood lymphocytes (PBLs) of normal controls and patients with Hodgkin's disease or non-Hodgkin's lymphoma and compared these values with those of Epstein-Barr virus (EBV)-transformed cell lines prepared from the same PBL samples. PBLs have an AGT level characteristic of the individual from whom the cells were obtained. The AGT activity of lymphoblastoid cell lines obtained from a control group of PBLs was significantly correlated with the activity of the PBLs from which they were derived (r = 0.742). There was no significant correlation between PBLs and EBV-transformed lines derived from these PBLs in Hodgkin's disease/non-Hodgkin's lymphoma patients (r = 0.407, -0.225, and 0.270 for patients prior to, during, or after therapy, respectively). The lack of significant correlation between lines and PBLs was not due to random fluctuations in AGT activity, because multiple lines prepared from the same PBL sample were found to be highly correlated in AGT activity. In order to account for these results, we suppose that PBLs from a given individual are a heterogeneous population with respect to AGT activity. In normal individuals, the AGT activity of early passages of the multi-clonal EBV-transformed cell lines reflect the AGT activity of the PBLs from which they were derived. Malignancy and/or treatment with chemotherapeutic agents may selectively affect those lymphocytes which are targets for EBV-transformation so that the resultant cell line is no longer representative (with respect to AGT activity) of the total PBL population. Long term culture of lymphoblastoid cell lines results in changes in AGT activity in some but not all cell lines suggesting that with time in culture, subsets with different AGT activities may be selected. There appears to be no growth advantage of low AGT activity and only rarely have we obtained lines with no measurable AGT activity, even after long periods in culture. PMID- 2548720 TI - Decreased activation of carcinogens in the liver of carcinogen-resistant rats. AB - We have developed a new strain of rats (resistant rat) which exhibit resistance against the carcinogenic action of several carcinogenic compounds. In the present study, we compared the ability of resistant rat liver to activate 3'-methyl-4 dimethylaminoazobenzene and 2-acetylaminofluorene to highly reactive metabolites, which can covalently bind to DNA, with that of carcinogen-sensitive parent strain rats (sensitive rat), using in vitro DNA binding assay. The covalent binding of these carcinogens to calf thymus DNA, catalyzed by the hepatic 9000 x g supernatant fraction from resistant rats pretreated with 3-methylcholanthrene, was about one-half of that catalyzed by the 9000 x g supernatant from sensitive rats which had received the same treatment. These results suggest that the ability of resistant rat liver to activate the carcinogens decreases compared to sensitive rat liver. Further experiments revealed that this decreased activation of the carcinogens in resistant rat livers is due to a low inducibility of 3 methylcholanthrene-inducible forms of cytochrome P-450 mRNA. The hepatic cytosolic Ah receptor concentrations in resistant rats were shown to be significantly lower than those of sensitive rats. Scatchard plot analysis demonstrated, however, that there is no significant difference between the affinity of Ah receptors for 2,3,7,8-tetrachlorodibenzo-p-dioxin in these two strains. These data implicate that the hepatic Ah receptor level may be an important factor in determining carcinogen sensitivity in rats. PMID- 2548721 TI - Identification of tissue factor in two human pancreatic cancer cell lines. AB - We have studied the effects of two human pancreatic cancer and two human small cell lung cancer cell lines on clotting and platelet aggregation. Both pancreatic lines markedly shortened recalcification times and induced platelet aggregation. The lung cancer lines produced little shortening of recalcification times and no platelet aggregation. The clotting and aggregation activities of the pancreatic lines were further characterized. Recalcification times following the addition of cancer cell line material to plasmas deficient in factors VII and X were markedly prolonged, suggesting that the activity is due to tissue factor. Hirudin, an inhibitor of thrombin from the saliva of leeches, and rabbit polyclonal immunoglobulin G anti-bovine brain tissue factor inhibited both procoagulant and aggregation activities. Apyrase (an enzyme degrading ADP), diisopropylfluorophosphate (a serine protease inhibitor) and L-trans epoxysuccinylleucylamido(4-guanidino)butane (a cysteine protease inhibitor) failed to inhibit these activities. Increasing concentrations of heparin inhibited platelet aggregation. Subcellular fractionation studies showed these activities to be localized to the plasma membrane. The association between mucin and the acceleration of clotting has been well described. The absence of mucin in electron micrographs of these pancreatic whole cells, membrane fractions, and shed microvesicles, as well as the failure of chaotropic agents (i.e., agents stripping material extrinsic to the cell membrane such as mucin) to abrogate this activity support these activities being intrinsic to the plasma membrane. These data strongly suggest that these activities are due to tissue factor which appears to be released as microvesicles in vitro. The release of tissue factor via microvesicles in vivo is one possible mechanism for the coagulopathy sometimes seen in patients with pancreatic carcinoma. PMID- 2548722 TI - Imaging necrotic myocardium: detection with 99mTc-pyrophosphate and radiolabeled antimyosin. AB - The major value of hot-spot imaging of the myocardium is its ability to define areas of necrosis rather than areas of diminished blood flow or cellular function. Applications of hot-spot imaging include the diagnosis and quantitation of myocardial infarction, myocarditis, and cardiac transplant rejection. The two agents in clinical use, 99mTc-Pyrophosphate and radiolabeled antimyosin, are discussed. PMID- 2548723 TI - Antibacterial activity of some plaque-disclosing agents and dyes. PMID- 2548724 TI - Sites and mechanisms of Ca2+ movement in non-excitable cells. AB - The level of free cytosolic Ca2+ ([Ca2+]i) in cells is firmly established as a second messenger alternative to the cyclic nucleotides. Regulation of the activity of Ca2+ requires the use of membrane transporters of various types which can be classified in terms of their transport rate; channels (fast), carriers (intermediate) and pumps (slow). In general channels are used to elevate [Ca2+]i whereas pumps decrease [Ca2+]i. At physiological membrane potential and Na+ gradients, carriers such as the 3Na+/Ca2+ exchanger also deplete the cell of Ca2+. The carriers could also function in a reverse mode especially with plasma membrane depolarization. Intracellular organelles which can incorporate Ca2+ from and return Ca2+ to the cytosol play a central role in determining [Ca2+]i in resting and stimulated cells. In the resting cell they function as the major Ca2+ buffering system while in the stimulated cell they participate in the dynamic control of [Ca2+]i. The collection of papers in this volume discusses the mechanisms of modulation of cell Ca2+ by these organelles. PMID- 2548725 TI - Mammalian phosphoinositide-specific phospholipase C isoenzymes. AB - Procaryotic and eucaryotic cells have evolved multiple pathways for communication with their external environment. The inositol 1,4,5-trisphosphate/diacylglycerol second messenger system is an example of such a signal transduction pathway which is present in multicellular eucaryotic organisms. Binding of an agonist to a specific cell surface receptor promotes rapid hydrolysis of phosphatidylinositol 4,5-bisphosphate. The pivotal enzyme for this second messenger system is phosphoinositide-specific phospholipase C which hydrolyzes phosphatidylinositol 4,5-bisphosphate to generate the two second messengers, inositol 1,4,5 trisphosphate and diacylglycerol. Recently, much progress has been made in the purification, characterization and cDNA cloning of multiple PI-PLC isoenzymes. The results of the recent studies on phosphoinositide-specific phospholipase C are reviewed. PMID- 2548726 TI - Modulation of intracellular free Ca2+ concentration by IP3-sensitive and IP3 insensitive nonmitochondrial Ca2+ pools. AB - Intracellular Ca2+ pools play an important role in the adjustment of cytosolic free Ca2+ concentrations. This review summarizes the recent knowledge on receptor mediated Ca2+ release and Ca2+ uptake mechanisms in Ca2+ stores of exocrine cells taking the exocrine pancreas and the parotid gland as an example. The intracellular mediator for agonist-induced Ca2+ release is inositol 1,4,5 trisphosphate (IP3) which acts by opening Ca2+ channels from the endoplasmic reticulum or a more specialized organelle called 'calciosome'. This Ca2+ release is the major event to increase cytosolic free Ca2+ concentrations of exocrine glands from a resting level of approximately 10(-7) mol/l to approximately 10(-6) mol/l. Subsequently also Ca2+ influx from the extracellular fluid into the cell is increased which involves the action of inositol 1,3,4,5-tetrakisphosphate (IP4). Intracellular nonmitochondrial Ca2+ reuptake occurs into IP3-sensitive (IsCaP) as well as into IP3-insensitive Ca2+ pools Ca2+ pools (IisCaP). While Ca2+ uptake into the IisCaP is mediated by a vanadate-sensitive Ca2+ pump, Ca2+ uptake into the IsCaP is mediated by a Ca2+/H+ exchanger at the expense of an H+ gradient which is established by a vacuolar type H+ pump present in the same Ca2+ pool. During stimulation both Ca2+ pools, IsCaP and IisCaP, are probably connected, the nature of which has not yet been clarified. It is suggested that GTP and/or IP4 control Ca2+ conveyance between intracellular Ca2+ pools by forming Ca2+-carrying junctions between membranes. Other models propose that Ca2+, which is released by IP3, induces Ca2+ release from another Ca2+ pool. Taking into account that H+ transport is present in IP3-sensitive Ca2+ pools the possibility of pH-regulated Ca2+ channels in the IisCaP, located in close neighbourhood to the IsCaP, is also considered. PMID- 2548727 TI - Isolation and functional characterization of an inositol trisphosphate receptor from brain. AB - We have identified an IP3 receptor protein in brain membranes through the binding of radiolabelled IP3. Autoradiographic studies localize the receptor to various areas of the brain with highest densities in Purkinje cells of the cerebellum. IP3 binding is inhibited by physiologic intracellular concentrations of calcium. Purification of the IP3 receptor to homogeneity reveals it to be comprised of four identical subunits of 260 kD each. Antisera to the purified receptor protein have been employed for immunohistochemical studies which, at the electron microscopic level, localize the IP3 receptor to a subdivision of the rough endoplasmic reticulum occurring in synaptic areas and in close association with the nuclear membrane. The IP3 receptor protein is selectively phosphorylated by cyclic AMP (cAMP) dependent protein kinase. This phosphorylation decreases 10 fold the potency of IP3 in releasing calcium from brain membranes. PMID- 2548728 TI - Calciosome, a sarcoplasmic reticulum-like organelle involved in intracellular Ca2+-handling by non-muscle cells: studies in human neutrophils and HL-60 cells. AB - Calciosomes are intracellular organelles in HL-60 cells, neutrophils and various other cell types, characterized by their content of a Ca2+-binding protein that is biochemically and immunologically similar to calsequestrin (CS) from muscle cells. In subcellular fractionation studies the CS-like protein copurifies with functional markers of the inositol 1,4,5-trisphosphate (IP3) releasable Ca2+ store. These markers (ATP-dependent Ca2+-uptake and IP3-induced Ca2+-release) show a subcellular distribution which is clearly distinct from the endoplasmic reticulum and other organelles. In morphological studies, antibodies against rabbit skeletal muscle CS protein specifically stained hitherto unrecognized vesicles with a diameter between 50 and 250 nm. Thus both, biochemical and morphological studies indicate that the calsequestrin containing intracellular Ca2+-store, now referred to as the calciosome, is distinct from other known organelles such as endoplasmic reticulum. Calciosomes are likely to play an important role in intracellular Ca2+-homeostasis. They are possibly the intracellular target of inositol 1,4,5-trisphosphate and thus the source of Ca2+ that is redistributed into the cytosol following surface receptor activation in non-muscle cells. PMID- 2548729 TI - Cellular dialogs during development. PMID- 2548730 TI - Neural induction is mediated by cross-talk between the protein kinase C and cyclic AMP pathways. AB - Embryonic inductions appear to be mediated by the concerted action of different inducing factors that modulate one another's activity. Such modulation is likely to reflect interactions between the signal transduction pathways through which the inducing factors act. We tested this idea for the induction of neural tissue. We report that both adenylate cyclase activity and cAMP concentration increase substantially in induced neuroectoderm during neural induction. The enhancement of adenylate cyclase activity requires protein kinase C (PKC) activation, indicating cross-talk between these two signal transduction pathways. This cross talk appears to be essential for neural induction. Whereas cAMP analogs alone were not neural inducers, they had a synergistic inducing effect if ectoderm was first incubated with TPA (12-O-tetradecanoylphorbol 13-acetate), a PKC activator. These results strongly suggest that at least two signals mediate neural induction. The first signal activates PKC and the second signal then activates the cAMP pathway effectively. PMID- 2548732 TI - The combined action of two intercellular signaling pathways specifies three cell fates during vulval induction in C. elegans. AB - Each of the six C. elegans vulval precursor cells (VPCs) has three potential fates (1 degree, 2 degrees, or 3 degrees). The fate of each VPC depends on two types of signals: a graded inductive signal that acts at a distance and a short range lateral signal among the VPCs. We describe interactions among mutations that cause different misspecifications of VPC fates. Particular combinations of mutations cause all six VPCs to have a single fate independent of their positions. Our results suggest that specification of the three VPC fates is accomplished by two binary decisions, each effected by one of the two signaling pathways. The gene lin-12 acts in the lateral signaling pathway and specifies 2 degrees. The "vulvaless" and "multivulva" genes act in the inductive signaling pathway and specify 1 degree independently of lin-12 and 2 degrees via lin-12. We describe a model for the regulatory circuitry underlying VPC determination that includes a role for lin-12 in both autocrine and paracrine VPC signaling. PMID- 2548731 TI - Transformation by activated ras or fos prevents myogenesis by inhibiting expression of MyoD1. AB - Transformation of myoblasts by activated ras inhibits myogenic differentiation. We demonstrate that this oncogene inhibits expression of the muscle regulatory factors MyoD1 and myogenin. Expression of retroviral-encoded MyoD1 in ras transformed myoblasts leads to the re-expression of both terminal differentiation markers and lineage markers expressed in proliferating myoblasts (including endogenous MyoD1 and myogenin), suggesting that ras inhibits myogenic differentiation in a manner dependent on the loss of MyoD1 expression. In addition, we show that fos transformation of myoblasts inhibits muscle differentiation by a similar mechanism. PMID- 2548733 TI - SecB functions as a cytosolic signal recognition factor for protein export in E. coli. AB - A purified 64 kd protein, consisting of four identical subunits of the 16 kd SecB, binds to the signal sequence of preproteins prior to their translocation across inverted vesicles (INV) derived from the E. coli plasma membrane. The purified SecB tetramer competes with canine signal recognition particle (SRP) in signal sequence binding and thus behaves as a prokaryotic equivalent of SRP. As shown by cell fractionation and immunoblot analysis with anti-SecB antibodies, SecB is a cytosolic protein. An E. coli supernatant depleted of SecB after passage through an anti-SecB Sepharose column retains full translation activity but is unable to support translocation into added INV. Translocation into INV is fully restored by readdition of purified SecB. PMID- 2548734 TI - Essential large transcripts of the maize Spm transposable element are generated by alternative splicing. AB - We used in vitro mutagenesis and cDNA cloning to identify new Suppressor-mutator (Spm) transposable element genes. Frameshift mutations in the ORFs of the tnpA gene's first intron markedly reduce Spm activity in transgenic tobacco, indicating that intron sequences encode essential gene products. Evidence is given that Spm encodes large alternatively spliced transcripts, designated tnpB (4.9 kb), tnpC (5.7 kb), and tnpD (5.8 kb), comprising all of the tnpA exons, most of the tnpA intron 1 ORF1 sequence, and either none, part, or all of the intron 1 ORF2 sequence. Two alternative splice donor sites were identified at the end of exon 1, and the structure of the different exon 1 sequences suggests that Spm employs a novel mode of translational regulation. PMID- 2548735 TI - An enhancer stimulates transcription in trans when attached to the promoter via a protein bridge. AB - Two principal models have been invoked to explain transcriptional stimulation of RNA polymerase II genes by enhancers/upstream promoter elements: in one, upstream regulatory sequences directly interact with proximal promoter elements via proteins bound to the DNA ("looping" model); in the other, RNA polymerase II (or a transcription factor) binds to distal sequences and then scans along the DNA until it reaches the promoter ("scanning" or "entry site" model). So far, it has been reported that enhancers or upstream promoter elements transmit their effect on a gene only via covalently closed DNA, i.e., in a cis configuration. The looping model predicts, however, that the effect can be transmitted also in certain trans configurations. Here we demonstrate that an enhancer from SV40 or cytomegalovirus can stimulate transcription in vitro even when noncovalently attached to the beta-globin promoter via the proteins streptavidin or avidin. These findings are consistent with the looping model rather than the scanning model. In addition, stimulation of transcription in trans, as shown by our experiments, may be found in nature in phenomena such as transvection, where one chromosome affects gene expression in the paired homolog. PMID- 2548736 TI - Site-specific recombination by Tn3 resolvase: topological changes in the forward and reverse reactions. AB - Site-specific recombination catalyzed by Tn3 resolvase proceeds with a linkage change, delta Lk, of +4 in the forward resolution reaction and -4 in the catenane fusion reverse reaction. The reverse reaction occurs only at low superhelical densities and gives unknotted circular products, consistent with plectonemic and not solenoidal wrapping of the two recombination sites. The strand exchange topologies are consistent with a mechanism in which resolvase cleaves all four DNA strands and religates them after a 180 degrees rotation of two duplex partners in a right-handed sense for the "forward" reaction, and in a left-handed sense for the "reverse" action. This could be achieved by a 180 degrees rotation of two resolvase subunits within a tetramer with D2 symmetry; we suggest that a different symmetry applies to phage lamda integrase catalysis. PMID- 2548737 TI - Viable deletions of a telomere from a Drosophila chromosome. AB - Destabilization of a P element transposon inserted in the subtelomeric region induced a set of similar chromosomal rearrangements. These rearrangements appear to be terminal deletions with endpoints clustered at the centromere-distal end of the transposon. The terminally deleted chromosome progressively loses sequences from the broken end at a rate of approximately 50-100 bp per fly generation, suggesting that the replication of this end may be incomplete. In most cases, capping of the broken end by readdition of new sequences was not observed. Past failures to recover terminal deletions of Drosophila chromosomes following X-ray mutagenesis may have been due to a cell cycle arrest in response to unrepaired DNA damage rather than to an absolute requirement for the telomere. PMID- 2548738 TI - A flow cytometric and immunofluorescence microscopic study of tumor necrosis factor production and localization in human monocytes. AB - The production and localization of tumor necrosis factor (TNF) in human monocytes were investigated by using monoclonal and polyclonal antibodies against recombinant human TNF together with flow cytometry and immunofluorescence microscopy. Lipopolysaccharide (LPS) induced a rapid and transient accumulation of TNF in perinuclear vesicles which was detected 20 min after the addition of LPS. The fluorescence intensity of the vesicles peaked at 40 min of LPS exposure, concomitantly with the release of TNF into the medium. Thus, our results indicate that the secretion of TNF is typical for secretory proteins as it involves passage through the secretory apparatus. Additional studies demonstrated that plasma membrane-associated TNF could not be detected in live monocytes not exposed to LPS. However, after 90 min with LPS, a small population of monocytes expressed membrane-associated TNF, and by 24 hr approximately 50% of the monocytes displayed TNF on the plasma membrane. Furthermore, our results indicate that plasma membrane-associated TNF does not represent released TNF bound back to its own receptor. Thus, our findings support the view that TNF exists as a surface trans-membrane protein in LPS-stimulated monocytes. PMID- 2548739 TI - Potentiation of the proliferative response of human B lymphocytes to low molecular weight B cell growth factor (LMW-BCGF) by fibroblast growth factors (FGFs). AB - Both acidic and basic fibroblast growth factor (FGF), although devoid alone of growth-promoting ability on resting or activated human lymphoid B cells, were found to markedly increase the proliferative response of anti-mu-chain or SAC preactivated B cell blasts to the low molecular weight B cell growth factor (LMW BCGF) and to enhance the costimulatory response of resting B cells to anti-mu chain and LMW-BCGF. This potentiating effect was also observed for a LMW-BCGF dependent B cell tumor derived from a lymphocytic nodular lymphoma. Other growth factors acting on fibroblasts, such as epidermal growth factor, alpha-thrombin, platelet-derived growth factor, and insulin-like growth factor-I did not display such enhancing effect on LMW-BCGF-driven proliferation. Activated, but not resting B cells were found to bear receptor sites for FGFs and from kinetics experiments, it is suggested that LMW-BCGF induces competence expression for FGFs in those cells. Moreover, the LMW-BCGF-elicited generation of inositoltrisphosphate resulting from polyphosphoinositides hydrolysis was increased in the presence of FGF. PMID- 2548740 TI - Differential interleukin-2 and interferon-gamma production by human lymphocyte cultures exceptionally resistant to Epstein-Barr virus immortalization. AB - Epstein-Barr virus (EBV) readily immortalizes human peripheral blood lymphocytes (PBL) in vitro. However, during the past several years, we found that PBL from two exceptional EBV-seropositive healthy adult individuals were refractory to immortalization by EBV. We report here a study aimed at learning about the immunobiological features which differentiate these EBV-resistant (R) PBL from others which are susceptible (S) to EBV immortalization. Results of this investigation indicate that: (a) Following EBV infection, R-PBL produced significantly higher amounts of interferon gamma (IFN-gamma) than S-PBL. There were however no differences in regard to interferon alpha production between these two types (R and S) of EBV-infected cultures. (b) R-PBL had a maximal interleukin-2 (IL-2) production by S-PBL occurred at least 48 hr later, i.e., at Day 7. (c) The percentage of non-B cells expressing the IL-2 receptor was also higher in EBV-infected R-PBL than S-PBL. (d) In contrast, expression of IL-2 receptors after EBV infection was higher on B cells from S-PBL than on B cells from R-PBL. Interestingly, no differences were noted in regard to IL-2 receptor expression between R-PBL and S-PBL treated with mitogens (i.e., phytohemagglutinin and pokeweed mitogen). (e) Finally, using anti-IL-2 and anti IFN-gamma antibodies in EBV-infected R-PBL cultures, we were able to obtain EBV induced immortalization of these cultures. Taken together, these results suggest that an early IL-2 synthesis and high IFN-gamma production by EBV-infected PBL play an important role against lymphocyte immortalization by EBV. PMID- 2548741 TI - ESR studies on active oxygen radicals produced in the respiratory burst of human polymorphonuclear leukocytes. AB - The mechanism and process of production of active oxygen radicals in the respiratory burst of polymorphonuclear leukocytes (PMN) stimulated with PMA (phorbol myristate acetate) was studied in this paper. The experimental results indicate that when the PMA was dilute enough or at the beginning of stimulation even when the PMA concentration was high, the spectrum of hydroxyl radical spin adducts, DMPO-OH, was dominant in the ESR spectra. However, at the maximum level of the respiratory burst, the spectrum of superoxide anion spin adducts, DMPO OOH, was dominant. PMID- 2548742 TI - Myosin regulation and calcium transients in fibroblast shape change, attachment, and patching. AB - Following our study in Balb/c 3T3 cells and other cultured fibroblasts of the changes in myosin light chain phosphorylation associated with alterations in cell shape, attachment, and receptor patching, we have now determined the corresponding changes in cytoskeletal myosin distribution, and in the cellular calcium concentration, since this might, in part, mediate such responses. Immunofluorescence microscopy showed that myosin assembly into ordered forms such as actomyosin bundles and myosin sheath almost always correlated with previously shown high phosphorylation levels of myosin regulatory light chain, whereas diffuse distributions usually correlated with low or undetectable levels. An exception was observed in treatment to alter cellular cAMP levels when, in a biphasic response, assembly was correlated inversely with the phosphorylation states shown previously. Fluorescent indicators for intracellular calcium concentration, [Ca++]i, showed that myosin disassembly by trypsin or EGTA acting externally on the cells was preceded by a transient increase in [Ca++]i. For EGTA this was associated with transient recruitment of myosin into dorsal sheath structure as well as the transient enhancement of phosphorylation shown earlier. Blockage of EGTA-induced disassembly could be achieved by azide, which also caused an immediate increase in [Ca++]i and inhibited its subsequent decline. Trypsin-induced dephosphorylation did not appear to involve an eventual reduction of [Ca++]i. Therefore, in many but not all of the systems studied, correlated changes were observed in myosin assembly, [Ca++]i, and the myosin phosphorylation levels shown earlier. PMID- 2548743 TI - Identification in maize mitochondrial 26S rRNA of a short 5'-end sequence possibly involved in transcription initiation and processing. AB - By direct RNA mapping, we have identified the precise 5' end of the maize mitochondrial 26S rRNA. The 5' termini of the 26S rRNA are 17 and 18 nt downstream from the 5' end predicted by Dale et al. (1984) from homology between the maize rRNA and E. coli large subunit ribosomal RNA. In addition, we have discovered a larger 26S species, presumably a 26S precursor, and precisely mapped its 5' end. The maize mitochondrial rRNA genes are probably regulated by control regions that differ from those already characterized in other genomes: the maize mitochondrial 26S, 18S and 5S rRNAs lack sequences that resemble the promoter regions of genes from other mitochondria and bacteria. However, the mature and precursor rRNAs all contain a tetranucleotide, AAUC, at their 5' ends (AAAC in the 5S rRNA). The sequence is also seen in the same position in several other plant mitochondrial mRNAs. We propose therefore that AAUC is a transcription or processing signal which is possibly unique to plant mitochondria. PMID- 2548744 TI - [The trial preparation of attenuated live hepatitis A vaccine]. AB - The experimental attenuated live hepatitis A vaccine has been prepared in KMB17 human fetal lung diploid cells with the HAV variant H2M20K5 (32 degrees C). No extraneous agent was detected in the vaccine, which had a titer of 10(6.5) TCID50/ml. Twelve monkeys vaccinated intravenously or subcutaneously were found to have no significant clinical hepatitis but with anti-HAV antibody in all cases. Thus the safety and immunogenicity of the vaccine were evidenced in monkeys and it is necessary to investigate the trial vaccination further in a few susceptible individuals. PMID- 2548745 TI - [Studies on Epstein-Barr virus strain differences with monoclonal antibodies]. AB - In order to differentiate a new EBV strain, H18, from other EBV strains, we prepared and screened by indirect immunofluorescence (IF) viable cells and fixed cells of a series hybridoma lines that produce monoclonal antibodies against H18 EBV membrane antigen (MA). T1-11 clone was used for further study: T14-11 antibody neutralizes EBV in vitro and recognizes polypeptides P220, P175 and P140 from purified H18 EBV particles, as shown by the Western Blot technique. The presence of the three polypeptides on the plasma membrane and in the cytoplasm may be detected by IF using T14-11 antibody. Treatment with puromycin completely abolished the expression of the antigen recognized by T14-11, and this Ab was shown to be specific for an early membrane antigen of EBV. PMID- 2548746 TI - [Clinical study of allergic rhinitis treated by "bu qi gu biao" therapy]. AB - The authors summarized 500 cases of allergic rhinitis treated by "Bu Qi Gu Biao" therapy since 1980. After treatment, long term curative effect was 87%. Moreover, the authors had observed serum IgE, IgA and IgG, plasma cyclic nucleotides, blood flow in nasal mucosa, and ultrastructure of nasal mucosa before and after the treatment in 200 cases. The results indicated that "Bu Qi Gu Biao" therapy could adjust immune function and cyclic nucleotides metabolism in organism, improve blood flow in nasal mucosa tissue, restore or improve cell's shape and function and eliminate partial deposition of immune compound in vascular basement membrane. Therefore, it could control or eliminate the symptoms of allergic rhinitis. PMID- 2548747 TI - Fluorescence correlation spectroscopy for detecting submicroscopic clusters of fluorescent molecules in membranes. AB - The formation of cell surface receptor clusters has been implicated or confirmed in the mechanism of signal transduction across biological membranes for a variety of processes, including receptor-mediated phagocytosis and endocytosis and cellular response to hormones and neurotransmitters. Fluorescence correlation spectroscopy (FCS) is one technique that may provide insight into the kinetics and extent of receptor aggregation. Recent theoretical and experimental developments in FCS for the investigation of submicroscopic clusters of fluorescent molecules are described and the potential applications of the technique to receptor aggregation are reviewed. PMID- 2548749 TI - High affinity diethylnitrosamine-deethylase in liver microsomes from acetone induced rats. AB - The effects of acetone treatment on microsomal cytochrome P-450-dependent mono oxygenases of the rat liver have been investigated to elucidate the role of this system in the metabolism of diethylnitrosamine (DEN). Acetone markedly enhanced the hepatic P-450 content and the activities of p-nitrophenol hydroxylase, acetone hydroxylase, ethoxycoumarin deethylase and DEN deethylase (DENd), whereas activities of pentoxy-resorufin O-deethylase and ethoxy-resorufin O-deethylase were not affected. Two distinct apparent Km values (0.43 and 9.1 mM), dependent on the substrate concentration, were observed for the DENd of acetone-induced microsomes. Only one Km value (8.4 mM) was observed for the DENd of control microsomes. In control microsomes at a DEN concentration of 1 mM, the N deethylation of DEN was undetectable whereas in acetone-induced microsomes the N deethylation rate was approximately 2.3 nmol/mg protein per min. The results suggest that acetone-induced microsomes of rat liver contain a high affinity form of DEN-deethylase which should be the P-450j isozyme (known to catalyze the oxidation of dimethylnitrosamine at low Km). P-450j is strongly enhanced by acetone treatment as indicated by the increase of the specific acetone hydroxylase. The treatment also enhanced the metabolism of DEN at substrate concentrations higher than 1 mM, suggesting that other P-450(s) catalyse DEN deethylation although with lower substrate affinity. The low Km form of DENd is a P-450-dependent mono-oxygenase. It requires NADPH and O2, is inhibited by CO, but not by mannitol, superoxide dismutase, catalase or desferrioxamine. Its action therefore appears not to be mediated by oxygen radical species. Many solvents such as dimethylsulfoxide, dioxolane, chloroform and butanol when present at 10 mM in the incubation mixture inhibited the low Km form of DENd. However, pyrazole and piperonylbutoxide were found to be the strongest inhibitors. These results establish that acetone affects the metabolism of DEN, particularly at low concentrations, in a fashion somewhat similar to dimethylnitrosamine. PMID- 2548748 TI - In vitro DNA modification by the ultimate carcinogen of 4-nitroquinoline-1-oxide: influence of superhelicity. AB - The effect of DNA tertiary structure on in vitro modification by 4-acetoxy aminoquinoline-1-oxide (Ac-4-HAQO) was investigated. The reactivity of pAT153 plasmid DNA depended on the conformational state of the molecule: it progressively decreased according to the decrease of the superhelical tension, being negatively supercoiled DNA about two times more susceptible than singly nicked relaxed DNA. HPLC of the three main Ac-4-HAQO adducts showed that 3 (deoxyguanosin-N2-yl)-4-aminoquinoline-1-oxide, N-(deoxyguanosin-C8-yl)-4 aminoquinoline-1-oxide and 3-(deoxyadenosin-N6-yl)-4-aminoquinoline-1-oxide accounted for 50, 25 and 10% of total quinoline DNA base adducts in all DNA conformations tested, except in the negatively supercoiled topoisomers where they accounted for 80, 15 and 5% respectively. DNA modification by Ac-4-HAQO resulted also in the formation of apurinic/apyrimidinic sites and in strand scissions. The quantification of these damages revealed that they represent an important fraction of all damaging events and that their yield is also influenced by DNA superstructure. Thus, these lesions must be considered as important DNA damage induced by Ac-4-HAQO. PMID- 2548751 TI - N-acyl dehydroalanines protect from radiation toxicity and inhibit radiation carcinogenesis in mice. AB - N-Acyl dehydroalanines have shown free radical scavenging activity. They react with and scavenge mainly oxygen-derived free radicals such as the superoxide anion (O2-.) and the hydroxyl radical (HO.). Ortho-methoxyphenylacetyl dehydroalanine (AD-20) protects total-body irradiated mice against the toxicity induced by X-rays when delivered as a single dose of 700 rads in a short period of time. This degree of protection was of the same order of magnitude as that obtained with the aminothiol S-2-(3-aminopropylamino)-ethylphosphorothioic acid (WR-2721). The radioprotection of AD-20 is extended to all other doses of X-rays tested (from 600 to 800 rads). Furthermore, AD-20 inhibits the development of thymic lymphomas in C57Bl/Ka mice undergoing a leukaemogenic course of irradiation (4 x 175 rads applied at weekly intervals). We postulate that AD-20 may act as a radioprotector and anticarcinogenic agent, most probably by inactivating the oxygen-derived free radicals formed during water radiolysis. PMID- 2548750 TI - Co-carcinogenic effects of NaHCO3 on o-phenylphenol-induced rat bladder carcinogenesis. AB - The role of urinary pH and Na+ concentration on the bladder carcinogenesis of o phenylphenol (OPP) was examined in male F344 rats. The rats were given powdered diet containing 2% sodium o-phenylphenate (OPP-Na, group 1), 1.25% OPP plus 0.64% NaHCO3 (group 2), 1.25% OPP plus 0.32% NaHCO3 (group 3), 1.25% OPP plus 0.16% NaHCO3 (group 4), 1.25% OPP (group 5), 0.64% NaHCO3 (group 6) or no test chemical (group 7) for 104 weeks respectively. Incidences of bladder carcinoma induced were significantly higher in groups 1 (12 of 29 rats, 41.4%) and 2 (9 of 29 rats, 31.0%) than in group 7 (0 of 27 rats, 0%). Groups 3 and 4 induced bladder carcinomas in 4 of 29 rats (13.8%) and 4 of 26 rats (15.4%) respectively, whereas no tumors occurred in group 5 (0 of 27, 0%). The incidence in group 6 was 3.6% (1 of 28 rats). Groups 1 and 2 induced significant increases in urinary pH and Na+ concentrations, whereas group 5 did not. Groups 3 and 4 showed the same tendency as groups 1 and 2. Examination with a scanning electron microscope showed the appearance of pleomorphic microvilli, short, uniform microvilli, and ropy or leafy microridges on the luminal surface of the bladder in groups 1-5 of rats treated with OPP or OPP-Na for 8 weeks. The appearance and severity were the same in groups 1 and 2, followed by the groups with decreasing doses of NaHCO3. The results indicated that OPP-Na is carcinogenic for the rat bladder, but OPP is not. However, increased urinary pH and Na+ concentration play important roles in OPP-Na rat bladder carcinogenesis. PMID- 2548752 TI - Effects of urinary potassium and sodium ion concentrations and pH on N-butyl-N-(4 hydroxybutyl)nitrosamine-induced urinary bladder carcinogenesis in rats. AB - The promoting activities of low and high sodium or potassium ion concentrations, under conditions of neutral as well as elevated urinary pH, in urinary bladder carcinogenesis, were investigated in rats treated with N-butyl-N-(4 hydroxybutyl)nitrosamine (BBN). Male Wistar rats were given 0.05% BBN in their drinking water for 4 weeks and then treated for 32 weeks with either control diet (group 1) or this diet supplemented with equimolar amounts of the following minerals: 2.34% NaCl (group 2), 2.98% KCl (group 3), 3.36% NaHCO3 (group 4), 1.68% NaHCO3 + 2% KHCO3 (group 5), or 4% KHCO3 (group 6). The alkalizing salts NaHCO3 and KHCO3 induced comparable increases in urinary pH and elevated urinary sodium or potassium ion concentrations respectively. The combination of NaHCO3 + KHCO3 similarly caused an elevation of the urinary pH and less increased sodium and potassium ion concentrations. In the groups fed NaHCO3 and KHCO3 either alone or in combination, the incidences of papillary/nodular hyperplasia, papillomas and carcinomas in the urinary bladder had increased as compared to controls. NaCl and KCl also induced high urinary sodium or potassium ion concentrations without alteration of urinary pH. This was accompanied by increased incidences of simple hyperplasia, papillary/nodular hyperplasia, and/or papillomas but no carcinomas. The present results indicate that the potassium ion is as potent as the sodium ion in promoting urinary bladder carcinogenesis under conditions of elevated urinary pH, and that both the sodium and potassium ions may exert weak promoting activity under conditions of neutral urinary pH. PMID- 2548753 TI - Ring-opened 7-methylguanine nucleotides are resistant to nuclease P1 digestion and good substrates to polynucleotide kinase. AB - Dimethyl sulfate was used to prepare 7-methyl-2'-deoxy-guanosine 3'-monophosphate (7-methyl-dGMP), which was ring-opened in alkali to 2'-deoxy-N5-methyl-N5-formyl 2,5,6-triamino-4-oxopyrimidine 3'-monophosphate (ROM-dGMP). ROM-dGMP was not dephosphorylated by nuclease P1 in contrast to normal deoxynucleotides. It was efficiently 5'-phosphorylated by T4 polynucleotide kinase. When methylated DNA was alkali-treated and digested with micrococcal nuclease, spleen phosphodiesterase and nuclease P1, ROM-dGMP was formed and this was labeled with [gamma-32P]-ATP in the presence of polynucleotide kinase. Ring-opening and P1 treatment appear methods of choice for 32P-post-labeling of 7-alkylguanines in DNA. PMID- 2548754 TI - Isolation and characterization of parenchymal cells from experimentally induced macronodular rat liver cirrhosis. AB - Hepatocytes were isolated from thioacetamide (TAA)-induced macronodular cirrhotic rat livers by a collagenase perfusion method. In the content of cellular metabolites, fatty acid uptake and lipid secretion there were no substantial differences compared with cells isolated from micronodular cirrhosis described previously. In contrast to isolated hepatocytes from normal livers those from macronodular cirrhosis had a lowered cellular content of triglycerides, phospholipids and cholesterol but not of cholesterol esters and free fatty acids. In macronodular cirrhosis hepatocytes of hypertrophic type, rich in cell organelles, can be distinguished ultrastructurally from those with signs of atrophy and degeneration. Immediately after isolation many hepatocytes isolated from macronodular cirrhosis showed plasma membrane blebbing. Whereas the blebbing was without recognizable effects on the fine structure of the isolated hepatocytes of the hypertrophic type, in the more atrophic ones some mitochondria were swollen. In addition, morphological analysis of the crude and purified suspensions revealed a partial selection of the hypertrophic cells during the isolation procedure, presumably due to a more labile state of those cells which showed signs of atrophy and degeneration. When stabilized in the suspension medium, however, the hepatocytes maintained complex metabolic functions for at least 2 h. Thus, the method described allows the isolation of parenchymal cells from TAA-induced macronodular cirrhotic livers for studying ultrastructural and biochemical alterations in hyperregenerative experimental liver cirrhosis. PMID- 2548755 TI - Methotrexate: studies on cellular metabolism. III.--Effect on the transplasma membrane redox activity and on ferricyanide-induced proton extrusion by HeLa cells. AB - The effect of methotrexate (MTX) on transplasma-membrane electron transport and ferricyanide-induced proton extrusion by HeLa cells was studied. Both systems were inhibited by MTX. It is suggested that inhibition of electron transport and proton extrusion caused by MTX could be associated with other metabolic alterations such as response to the increase in NADH levels and decrease in intracellular pH. PMID- 2548757 TI - Metabolism of adenine nucleotides in human blood. AB - Biologically active concentrations of potently vasoactive and platelet-active adenine nucleotides are generated in plasma by a variety of pathophysiological mechanisms. Although there is evidence that ATP and ADP are inactivated by endothelial ectonucleotidases, there has been little attempt to study the metabolic routes of their catabolism in blood or to assess the contribution of this process to their clearance in vivo. Therefore, we have studied the rates and patterns of catabolism of ATP, ADP, and AMP in whole blood, plasma, and isolated blood cells. Rates of degradation of each nucleotide in cell-free plasma ranged from 0.07-0.32 nmol/min/ml with 1 microM substrates to 1.1-3.6 nmol/min/ml with 100 microM substrates. The pattern of catabolism indicated that sequential dephosphorylation from ATP----ADP----AMP----adenosine occurs. In whole blood, the pattern was similar although ATP and ADP (but not AMP) breakdown was more rapid. This was due to leukocyte ectonucleotidase activity. The use of selective inhibitors demonstrated that catabolism was not due to nonspecific phosphatase activity and that plasma 5'-nucleotidase is distinct from ATPase or ADPase. In leukocytes, ATPase and ADPase activities were distinguishable, and each contributed substantially to the rates of catabolism in whole blood. Leukocyte 5' nucleotidase did not measurably contribute to AMP dephosphorylation in blood. By comparison, ecto-ATPase and ecto-ADPase activities on cultured human umbilical vein endothelial cells were similar to those on leukocytes while endothelial 5' nucleotidase per 10(6) cells was equivalent to the soluble activity in 1 ml of blood or plasma.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548756 TI - Some effects of fructose-1,6-diphosphate on rat myocardial tissue related to a membrane-stabilizing action. AB - This study aims at elucidating the mechanism of action of extracellular fructose 1,6-diphosphate (FDP). FDP is able to inhibit Ca++ entry into the myocardial tissue with an IC50 value of 11.5 mM and in addition, it is bound by rat heart slices, the binding being activated by Zn and conditions of chemical hypoxia induced by KCN and iodoacetate. The overall effect of extracellular FDP includes an increase of frequency and amplitude of contraction of perfused heart at concentration below 1 mM, and, in general, a stimulation of the oxygen consumption of the tissue. The antihaemolytic effect of FDP suggests its action as a membrane stabilizer. The effects of extracellular FDP on the myocardial cell can be interpreted both on the basis of a limited permeability of the cell membrane to it and as a purely extracellular effect transduced through the cell membrane with a final response consisting of an increase in the intracellular FDP. PMID- 2548758 TI - Prostaglandin E receptors in cardiac sarcolemma. Identification and coupling to adenylate cyclase. AB - Purified cardiac sarcolemmal membrane vesicles were used to determine if specific prostaglandin (PG) receptors are present on the myocyte. Two binding sites for PGE2 were identified in isolated bovine sarcolemmal membranes: a high-affinity site with a dissociation constant (Kd) of 0.32 nM and a maximum binding (Bmax) of 376 fmol/mg of protein and a lower-affinity site with a Kd of 3.41 nM and a Bmax of 2,112 fmol/mg of protein. In competition experiments, unlabeled PGE1 displaced [3H]PGE2 from its membrane receptor at concentrations similar to those of unlabeled PGE2. Both PGF2 alpha and PGD2 displaced [3H]PGE2 from the membrane, but only at high concentrations (greater than 10(-6) M and greater than 10(-5)M, respectively). Digestion of sarcolemmal membrane with trypsin resulted in a threefold decrease in specific [3H]PGE2 binding. Phosphorylation of the membrane with protein kinase A also decreased specific [3H]PGE2 binding. At concentrations of PGE2 that occupy the high-affinity site, sarcolemmal adenylate cyclase activity was inhibited in the presence of 5'-guanylylimidodiphosphate [Gpp(NH)p]. We conclude that the isolated cardiac sarcolemmal membrane contains a high affinity binding site for PGE2 that is functionally coupled to adenylate cyclase. The binding site is stereospecific and probably recognizes the 9-keto,11-hydroxyl portion of the ring structure of these prostaglandins. PMID- 2548759 TI - Intracoronary injections of salbutamol demonstrate the presence of functional beta 2-adrenoceptors in the human heart. AB - To demonstrate the presence of functional cardiac beta 2-adrenoceptors in man, we studied the responses to intracoronary injections of salbutamol in three groups of six patients. We injected salbutamol, a selective beta 2-adrenoceptor agonist, into the right coronary artery to avoid peripheral vasodilator action and to stimulate the sinoatrial node directly. Salbutamol injections caused a sinus tachycardia. The same doses of salbutamol injected into the aortic root caused no change in heart rate, ruling out a systemic effect. The mean dose required to cause an increase in heart rate of 30 beats/min (IHR30) was 2.6 micrograms in the first group of six patients. In 12 other patients salbutamol was given after beta blockade to confirm the beta 2-selectivity of the responses. Doses of practolol (beta 1-selective blockade) and of propranolol (beta 1- and beta 2-blockade) that had equal beta 1-blocking activity were used. In six patients who were given practolol, the mean IHR30 dose was 2.1 micrograms. In six patients who were given propranolol, the mean IHR30 dose was significantly greater at 64 micrograms (p less than 0.001, practolol vs. propranolol). This study demonstrates that direct cardiac beta 2-adrenoceptor stimulation in man has a positive chronotropic effect. PMID- 2548760 TI - Calcium, its role in isoproterenol-stimulated atrial natriuretic peptide secretion by superfused rat atria. AB - The beta-adrenergic agonist isoproterenol stimulates immunoreactive atrial natriuretic peptide (IR-ANP) secretion by superfused rat atria in vitro. beta Adrenergic agonists alter the cellular handling of calcium, which culminates in a rise in the systolic calcium concentration. This is achieved by increasing calcium influx through voltage-dependent calcium channels and by increasing the storage pool of calcium in the sarcoplasmic reticulum (SR). We therefore asked the question whether isoproterenol-stimulated IR-ANP secretion was dependent on the protein kinase A-induced rise in systolic calcium or was due to a direct effect of protein kinase A activation. Isolated rat left atria paced at 3 Hz were superfused in vitro. IR-ANP secretion was determined by radioimmunoassay of timed collections of the superfusate. Superfusion with 0.1 microM isoproterenol or 0.5 mM dibutyryl cyclic AMP increased IR-ANP secretion twofold. Stimulated IR-ANP secretion was lowered to near baseline by lowering the buffer calcium concentration from 1.8 to 0.2 mM or by adding to the superfusate 10 microM nitrendipine (a calcium-channel blocker) or 1 microM ryanodine (an inhibitor of SR calcium release). Superfusion of nonbeating, electrically quiescent left atria with 0.1 microM isoproterenol failed to stimulate IR-ANP secretion. We conclude: 1) Isoproterenol-stimulated IR-ANP secretion is dependent on calcium influx through voltage-dependent calcium channels and on the release of calcium from the SR. 2) Enhanced calcium influx alone is not adequate to maintain isoproterenol stimulated IR-ANP secretion. 3) The SR appears to be the primary source of calcium for isoproterenol-stimulated IR-ANP secretion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548761 TI - Marked reduction of free radical generation and contractile dysfunction by antioxidant therapy begun at the time of reperfusion. Evidence that myocardial "stunning" is a manifestation of reperfusion injury. AB - Recent evidence suggests that postischemic myocardial dysfunction ("stunning") may be mediated by oxygen free radicals, but the exact time window during which the critical radical-mediated damage develops remains unknown. Furthermore, the evidence for the oxyradical hypothesis is indirect and, therefore, inconclusive. Thus, the potent and cell-permeable antioxidant N-(2-mercaptopropionyl)-glycine (MPG) was administered as an intra-coronary infusion (8 mg/kg/hr) to three groups of open-chest dogs undergoing a 15-minute coronary occlusion followed by 4 hours of reperfusion. In group I (n = 8), the infusion of MPG was started 15 minutes before occlusion and ended 2 hours after reperfusion; in group II (n = 9), MPG was started 1 minute before reperfusion and ended 2 hours thereafter; in group III (n = 10), MPG was started 1 minute after reperfusion and ended 2 hours and 15 minutes thereafter. Control dogs (group IV) (n = 10) received vehicle. Recovery of contractile function (assessed as systolic wall thickening) was equivalent in groups I and II, and in both groups it was substantially greater than in controls (p less than 0.005 at 4 hours). In contrast, in group III recovery of function was indistinguishable from controls. To determine whether the protection afforded by MPG was due to inhibition of free radical reactions, myocardial production of free radicals was directly assessed by intracoronary infusion of the spin trap alpha-phenyl N-tert-butyl nitrone (PBN). In control dogs (group VII, n = 6), radical adducts of PBN were released in the coronary venous blood after reperfusion, with a burst occurring in the first 5 minutes. MPG given as in group II (group V, n = 5) markedly suppressed myocardial production of PBN adducts (delta = -98% over 3 hours, p less than 0.01 vs. controls); this effect was evident immediately after reperfusion. MPG given as in group III (group VI, n = 5) also suppressed PBN adduct production (delta = -83% over 3 hours, p less than 0.025 vs. controls), but this effect was delayed. Hence, the radicals important in myocardial stunning appear to be those generated immediately after reperfusion. In vitro studies demonstrated that MPG is an exceptionally powerful scavenger of .OH (rate constant = 8.1 x 10(9) M-1 sec-1 by pulse radiolysis) but has no significant effect on .O2- (rate constant less than 10(3) M-1 sec-1), H2O2 (rate constant = 1.6 M-1 sec-1), or non-.OH-initiated lipid peroxidation, suggesting that removal of .OH is the major mechanism of the beneficial effects of MPG.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2548762 TI - Enkephalinase inhibition increases plasma atrial natriuretic peptide levels, glomerular filtration rate, and urinary sodium excretion in rats with reduced renal mass. AB - To investigate the in vivo effects of inhibition of endopeptidase 24.11, an enkephalinase enzyme shown to be involved in atrial natriuretic peptide (ANP) breakdown in vitro, we infused phosphoramidon, a specific inhibitor of endopeptidase 24.11, into rats with reduced renal mass (and chronic extracellular volume expansion) and into normal rats. Relative to baseline values in rats with remnant kidneys, phosphoramidon led to elevations of plasma ANP levels and concomitant increases in urinary sodium excretion, fractional excretion of sodium, glomerular filtration rate, filtration fraction, and urinary cyclic GMP excretion. Similar changes in renal function and urinary cyclic GMP excretion were obtained with thiorphan, another endopeptidase 24.11 inhibitor. These enhanced ANP levels and renal actions were not observed with phosphoramidon in normal rats. These results show that plasma ANP levels can be modulated in rats with reduced renal mass by inhibition of endopeptidase 24.11. PMID- 2548763 TI - Blockade of rabbit atrial sodium channels by lidocaine. Characterization of continuous and frequency-dependent blocking. AB - Lidocaine block of the cardiac sodium channel is believed to be primarily a function of channel state. For subthreshold potentials, block is limited to the inactivated state, whereas above threshold, block results from the combination of open- and inactivated-state block. Since, in the absence of drug, inactivation develops with time constants that vary from several hundred milliseconds to a few milliseconds as potential is varied from subthreshold to strongly depolarized levels, we would predict a similar voltage dependence of at least a fraction of block. Prior theoretical analyses from our laboratory suggest that there should be a direct parallel between blockade determined with a single pulse and trains of pulses. We tested these predictions by measuring the blockade of sodium current in cultured atrial myocytes during exposure to 80 microM lidocaine. We selected two test potentials for most of our studies, -80 mV, which was clearly in the subthreshold range of potentials, and -20 mV, which was close to the peak of the current-voltage curve. With single pulses of increasing duration, block developed with a single exponential time course and with time constants that decreased from 694 +/- 117 msec at -80 mV to 373 +/- 54 msec at -20 mV. In the absence of drug, inactivation developed with a time constant 176 +/- 17 at -80 mV and 2.9 +/- .5 msec at -20 mV. Despite the much slower onset of inactivation at 80 mV, no second-order delay in block development was observed. This suggests that at -80 mV block is occurring to a channel conformation that is accessed without delay rather than the classical inactivated state. We compared the kinetics of block during a single continuous pulse with trains of pulses at -20 mV. The rate of block onset was faster during the pulse trains, suggesting an element of "activated state" block. We computed shifts in apparent inactivation from observed steady-state blockade. The computed shifts agree well with those observed, indicating that shifts in apparent inactivation result largely from voltage-sensitive equilibrium blockade. The classical states described in the Hodgkin-Huxley formalism may be too restrictive to fully describe the voltage- and time-dependent block of cardiac sodium channels. PMID- 2548764 TI - IMP production by ATP-depleted adult rat heart cells. Effects of glycolysis and alpha 1-adrenergic stimulation. AB - A rapid deenergization procedure was used to probe the regulation of in situ adenylate deaminase and 5'-nucleotidase in isolated adult rat heart cells. In cells depleted of ATP, the rate of ionosine monophosphate (IMP) production was fourfold greater in cells that had been respiring prior to deenergization than in cells that had been maintaining ATP stores through anaerobic glycolysis. This effect of respiratory inhibition was fully reversed by reaeration. When phenylephrine was present during preincubation, IMP production during a subsequent 5-minute rapid deenergization was increased by 70% in respiring cells and by 88% in those that had not been respiring. These effects of phenylephrine were abolished by prazosin. Adenosine production by cells without ATP was inversely related to that of IMP, whereas it was positively correlated with the amount of AMP remaining in cells after 5 minutes. We conclude from these data that rat heart adenylate deaminase is regulated by a product(s) of anaerobic glycolysis and by alpha 1-adrenergic stimulation. The production of intracellular adenosine in cells without ATP, on the other hand, is governed primarily by the concentration of AMP and appears to be catalyzed by the cytosolic type I 5' nucleotidase. PMID- 2548765 TI - N-acetylcysteine potentiates platelet inhibition by endothelium-derived relaxing factor. AB - Recent evidence suggests that endothelium-derived relaxing factor exhibits properties of nitric oxide. Like nitric oxide, it inhibits platelet function and mediates its effects by elevating intracellular cyclic GMP. In this study we have investigated the role of reduced thiol in the mechanism of action of endothelium derived relaxing factor on platelets. Bovine aortic endothelial cells were grown on microcarrier beads and pretreated with aspirin before use. Endothelial cells stimulated with bradykinin or exposed to stirred medium expressed a dose dependent inhibition of platelet aggregation that was potentiated by the reduced thiol, N-acetylcysteine. Endothelial cell-mediated platelet inhibition was attenuated by methylene blue. Inhibition of platelet aggregation by endothelial cells was associated with a rise in platelet intracellular cyclic GMP, an effect that was enhanced by N-acetylcysteine. These data show that 1) the reduced thiol N-acetylcysteine potentiates platelet inhibition by endothelium-derived relaxing factor and 2) this effect is associated with increasing intracellular platelet cyclic GMP levels. PMID- 2548766 TI - Sodium-lithium exchange and sodium-proton exchange are mediated by the same transport system in sarcolemmal vesicles from bovine superior mesenteric artery. AB - Several laboratories have reported that Na+-Li+ countertransport activities are increased in red blood cells from patients with essential hypertension. It has been proposed that the activity of this red blood cell transport system might reflect the activity of a similar system in vascular smooth muscle. We previously demonstrated Na+-Li+ exchange in sarcolemmal vesicles from canine artery and proposed that this transport function might be mediated by the Na+-H+ exchanger. In the present studies, however, we were unable to demonstrate Na+-Li+ countertransport in canine red blood cells. Since bovine red blood cells have a vigorous Na+-Li+ exchanger and we previously demonstrated Na+-H+ exchange in sarcolemmal vesicles from bovine artery, we wished to determine whether bovine sarcolemmal vesicles mediate Na+-Li+ exchange and whether this transport function is mediated via the Na+-H+ exchanger. We found that an outwardly directed proton or Li+ gradient stimulated 22Na+ uptake in sarcolemmal vesicles from bovine superior mesenteric artery. Li+ gradient-stimulated Na+ uptake was not due to electrical coupling between the two ions, was not affected by a change in membrane potential, and could not be explained by the parallel operation of Li+ H+ and Na+-H+ exchange. External Li+ inhibited proton gradient-stimulated Na+ uptake, and external protons inhibited Li+ gradient-stimulated Na+ uptake. Na+ efflux from vesicles was stimulated by inwardly directed gradients for Li+ or protons, and these effects were not additive. Proton efflux from vesicles was stimulated by inwardly directed gradients for Na+ or Li+, and these effects were not additive. Finally, Na+-H+ exchange and Na+-Li+ exchange in sarcolemmal vesicles were inhibited by 5-(N-ethyl-N-isopropyl)amiloride in an identical dose dependent manner. In conclusion, Na+-Li+ countertransport could not be demonstrated in canine red blood cells, but as is the case with bovine red blood cells, sarcolemmal vesicles from bovine artery mediate Na+-Li+ countertransport. This transport function and sarcolemmal Na+-H+ exchange are mediated via a single 5-(N-ethyl-N-isopropyl)amiloride-sensitive cation exchanger with affinity for Na+, Li+, and protons. The cow, as opposed to the dog, may be a good animal model to test whether the activity of red blood cell Na+-Li+ countertransport is predictive of the activity of Na+-Li+ (and Na+-H+) exchange in vascular smooth muscle. PMID- 2548767 TI - [Report on a fungus parasitizing Entamoeba histolytica]. AB - Infection of Entamoeba histolytica with chytridiaceous fungus Sphaerita was observed in some specimens obtained from a farmer and stained with iron haematoxylin. The fungi were found in 78% of the cysts, mostly immature ones. Within the amoebae this parasite occurred singly, in groups, or in the form of a sporangium. It was located in the cytoplasm, the glycogen mass or the chromatoidal bars. In the same specimen, the parasitic fungus was also found in 18% of E. coli cysts; in 11% of E. nana cysts; while only one of 16 E. hartmanni cysts was parasitized. It is an interesting case of superimposed parasitism so far reported in China as well as a rare case of several species of amoebae being heavily involved with the same in the scientific literature. PMID- 2548768 TI - Long-term beta-blockade in dilated cardiomyopathy. Effects of short- and long term metoprolol treatment followed by withdrawal and readministration of metoprolol. AB - To evaluate the short- and long-term effects of beta-adrenergic blockade (metoprolol) as well as the reaction to withdrawal and readministration of metoprolol in severe heart failure, 33 patients (25 men and eight women; mean age, 47.6 +/- 14.0 years) with dilated cardiomyopathy were studied by right and left heart catheterization, right ventricular biopsy, two-dimensional and Doppler echocardiography, and external pulse recordings. Twenty-six of 33 patients survived more than 6 months, and 24 of the 26 patients improved their functional class (from mean 3.3 to 1.8, p less than 0.0001). These 24 patients were subjected to withdrawal of metoprolol until the number of symptoms increased and deterioration occurred as observed noninvasively (group 1, n = 16), whereas the eight patients did not deteriorate during a 12-month period (group 2). During long-term treatment with metoprolol, there was an increase in ejection fraction from 0.24 to 0.42 (p less than 0.0001), whereas there was a decrease in the left ventricular (LV) end-diastolic dimension (from 7.3 to 6.4 cm, p less than 0.0001), in the grade of mitral regurgitation (from 1.7 to 0.4, p less than 0.0001), and in the grade of tricuspid regurgitation (from 0.6 to 0.05, p less than 0.007). There was a decrease in pulmonary wedge pressure (from 23.8 to 10.7 mm Hg, p less than 0.0001), LV end-diastolic pressure (from 24.1 to 13.4 mm Hg, p less than 0.002), and systolic vascular resistance (from 1,782 to 1,499 dynes/sec/cm, p less than 0.04). There was an increase in systolic blood pressure (from 116 to 132 mm Hg, p less than 0.003), cardiac index (from 2.17 to 2.58 l/min/m2, p less than 0.005), and LV stroke work index (from 31 to 65 g.m/m2, p less than 0.0001). During withdrawal of metoprolol, the heart rate and left atrial dimension increased (p less than 0.0001), whereas ejection fraction decreased (p less than 0.0001). The 12 (of 16) patients in group 1 who survived the withdrawal period had metoprolol readministered, and subsequently, ejection fraction increased (from 0.23 to 0.33, p less than 0.002). Patients had a low number of ventricular beta-adrenergic receptors compared with healthy control subjects (30.3 +/- 2.9 vs. 97.4 +/- 8.7 fmol/mg protein, p less than 0.001), but long-term treatment with metoprolol caused a moderate up-regulation (from 30.3 +/ 2.9 to 49.0 +/- 7.1 fmol/mg protein, p less than 0.05), which may facilitate a more normal response to sympathetic stimulation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2548770 TI - Beta-adrenergic receptors on MNL. PMID- 2548769 TI - Regulation of cardiac beta-adrenergic receptors by captopril. Implications for congestive heart failure. AB - The interaction of the renin-angiotensin system and the sympathetic nervous system in patients with congestive heart failure is not well understood. We tested the hypothesis that angiotensin-converting enzyme inhibitors can resensitize the beta-adrenergic receptor system. Guinea pigs were given captopril, isoproterenol, or both for 2 weeks. At death, cardiac sarcolemmal and light vesicle fractions and intact mononuclear leukocytes were prepared. Captopril treatment led to an up-regulation of cardiac beta 1- but not mononuclear leukocyte beta 2-adrenergic receptors and an increase in isoproterenol-stimulated adenylate cyclase activity in the heart. Animals treated with isoproterenol developed cardiac hypertrophy, had increased plasma norepinephrine levels, and had a decreased number and responsiveness of both cardiac and mononuclear leukocyte beta-adrenergic receptors. Concomitant treatment with captopril attenuated alterations of heart weight, plasma norepinephrine levels, and cardiac beta-receptor density and function. In contrast to its cardiac effects, captopril treatment did not diminish the down regulation of mononuclear leukocyte beta 2-adrenergic receptors by isoproterenol. Our data suggest that captopril may resensitize the cardiac but not the mononuclear leukocyte beta-adrenergic receptor-adenylate cyclase system after long-term catecholamine exposure. PMID- 2548771 TI - Results of surgical management of nasopharyngeal angiofibroma. The Cleveland Clinic experience, 1977-1986. AB - During 1977 through 1986, 15 patients were evaluated and underwent surgery for nasopharyngeal angiofibroma at the Cleveland Clinic Foundation. All were males aged five to 20 years. Diagnoses were established clinically and radiographically. The most common presentations were nasal obstruction and epistaxis. Computed tomographic scans were obtained in almost all cases. In 14 patients, preoperative embolization of the tumor was performed at the time of diagnostic angiography; embolization of tumors made surgical excision easier and reduced blood loss. The most common surgical exposure used was a combination of transpalatal and transantral approaches. Follow-up of one to 10 years showed recurrence in only two patients. One had no further recurrence after subsequent surgery; the other remains asymptomatic with no further progression. The approach to diagnosis and treatment described results in cure for most patients. PMID- 2548773 TI - Serum arylesterase (paraoxonase) activity following myocardial infarction. PMID- 2548772 TI - Clinical biochemistry of neuron specific enolase. AB - The soluble brain protein 14-3-2 first described by Moore and McGregor in 1965 is now known to be a cell specific isoenzyme of the glycolytic enzyme enolase (EC 4.2.1.11), designated neuron specific enolase (NSE). It is not only a marker for all types of neurons, but also for all neuroendocrine or paraneuronal cells. The appearance of NSE is a late event in neural differentiation, thus making NSE a useful index of neural maturation. The demonstration that tumors of the nervous system and of neuroendocrine origin contain NSE has promoted the study of NSE as a possible tumor marker. Immunocytochemistry has been used to identify NSE in cytologic preparations from several types of tumors, offering useful indications for differential diagnosis. NSE levels in serum from tumor patients are not useful in the diagnosis of early stage disease. However, serum NSE levels have been shown to be helpful in the identification of advanced small cell lung cancer, neuroblastoma and several other neoplasms. The main use of serum NSE is the monitoring of chemotherapy and the detection of a relapse in these cases. PMID- 2548775 TI - Aarskog syndrome in Hungary. PMID- 2548774 TI - Caffeine as indicator of metabolic functions of microsomal liver enzymes. PMID- 2548776 TI - Expression and sequences of T cell receptor beta-chain variable genes in the enlarged lymph nodes of C57BL/6-lpr/lpr mice. AB - An autosomal recessive gene, lpr, is responsible for lymphoproliferation and autoimmunity of lpr-mice, in which background genes are also known to influence the development of autoimmune disease. To define the differences in abnormally proliferating T cells between C57BL/6-lpr/lpr and MRL/Mp-lpr/lpr mice, and to try and understand the influence of background in the differing expression of autoimmune disease in both strains, we analysed the sequences of T cell antigen receptor V beta genes expressed in the cells from the enlarged lymph nodes of C57BL/6-lpr/lpr mice. Eleven beta cDNAs out of the 38 C beta-specific cDNAs contained sequences with open reading frames from the beginning of the variable region to the expected termination codons at the end of the constant regions. Notably, 36% of the functional beta-chain mRNAs expressed V beta 8.3 genes, whereas V beta 8.1 and V beta 8.2 genes were not found. These results are consistent with a relatively lower frequency of the V beta 8.1 or V beta 8.2 expressing cells in the hypertrophic lymph nodes of C57BL/6-lpr/lpr mice, detected by KJ16-133 monoclonal antibody. Interestingly, other V beta genes expressed in these mice were completely distinct from those in MRL/Mp-lpr/lpr mice as described by Singer et al. (1986). The different distribution of V beta genes expressed in C57BL/6-lpr/lpr from that in MRL/Mp-lpr/lpr mice might be related to the differences in the genetic background and the expression of lpr gene-associated autoimmunity. PMID- 2548777 TI - Influence of donor characteristics on responsiveness of mononuclear cells to pokeweed mitogen. AB - The variability in response of peripheral blood mononuclear cells (PBMC) from 95 healthy donors to pokeweed mitogen (PWM) was investigated. Age, sex, and blood group of the donors, the numbers of PBMC recovered from 500 ml blood, the percentage of monocytes therein, the presence of antilymphocyte antibodies (L Cyt), or of anti-cytomegalovirus (CMV) antibodies, as well as HLA-B8 and HLA-DR4 were evaluated for their relation with the level of blast cell formation, proliferative response, and immunoglobulin G (IgG) synthesis, induced by PWM. The results showed that increasing age of the donors and the presence of anti-CMV antibodies are significantly associated with low proliferative responses of PBMC, whereas the HLA-B8 antigen and female donor sex were found to be associated with high blast cell formation after PWM stimulation. High percentages of monocytes in isolated PBMC at the onset of stimulation were inversely associated with IgG production, whereas the HLA-DR4 antigen was associated with high levels of IgG production, induced by PWM. The observed relations indicate that for comparison of responses of unseparated PBMC to PWM between patients and healthy controls, age, sex, and laboratory data, including the HLA background of donors, may have to be taken into account. PMID- 2548779 TI - Effect of Na+ pump suppression on reactivity of rat trachealis to cooling. AB - 1. This study tests the hypothesis that suppression of Na+ pump would increase the rate of tension development and the magnitude of contraction induced by cooling in airway smooth muscle. 2. Rat tracheal preparations were incubated in ouabain for 8 h and tested hourly for their response. In a representative specimen the rate of tension development increased from the control value of 0.7 mg/s to 5.5 mg/s after 7 h of incubation in ouabain concentration of 4 X 10(-4) mol/L; likewise, the magnitude of contraction increased from the control value of 80 mg to 550 mg. 3. Using ouabain concentrations between 9 X 10(-5) mol/L and 6 X 10(-3) mol/L, the rate of tension development and the magnitude of contraction first increased in a dose-dependent manner up to 8 X 10(-4) mol/L, then declined with higher doses but the responses were still greater than the control values at all concentrations. 4. After 3 h incubation in ouabain at 8 X 10(-4) mol/L, the mean rate of tension development and the mean magnitude of contraction increased to 647% and 578% of the control value, respectively. 5. These results indicate that depression of Na+ pump results in hypersensitivity and hyper-reactivity of the airway smooth muscle to cooling. PMID- 2548778 TI - Biochemical and hormonal changes during a 1000 km ultramarathon. AB - 1. To examine individual hormonal responses to extreme physical stress, blood samples were taken from eight highly trained athletes 1 day before and within 15 min of finishing the 1986 1000 km Sydney to Melbourne Ultramarathon foot race. 2. The baseline hormonal state of these highly trained athletes was quite different from normal. Resting serum conjugated catecholamines--epinephrine (E), norepinephrine (NE), dopamine (D), free E and free D--were significantly elevated above the normal mean (P less than 0.01). ACTH levels were significantly elevated above the normal range. Immunoreactive beta-endorphin (IR-beta EP), growth hormone (GH), prolactin (PRL), testosterone, cortisol and cortisol-binding globulin (CBG) were within the normal range. 3. The effect of the race on serum catecholamine levels was to elevate further free and conjugated NE (P less than 0.01). Other catecholamines, free and conjugated, remained significantly elevated above the normal mean (P less than 0.01). Adrenocorticotrophic hormone (ACTH) remained elevated, and IR-beta EP within the normal range, without significant change. A significant increase in GH (P less than 0.05), PRL (P less than 0.01), and cortisol (P less than 0.01) was seen, with no change in CBG. 4. As a model of chronic physical stress, the ultramarathon runner demonstrates a significantly altered baseline hormonal state as reflected in the primary mediators of the stress response, the catecholamines and the hypothalamic-pituitary-adrenal axis. Their response to severe exercise is distinct from that of untrained individuals in whom conjugated catecholamines decrease and ACTH increase. This may represent hormonal adaptation to prolonged stress. PMID- 2548780 TI - Effects of reserpine pretreatment on neuroeffector transmission in the vas deferens. AB - 1. The effects of reserpine pretreatment on neurotransmission in the guinea-pig vas deferens have been re-examined with a view to study the role of noradrenaline (NA) in mediating postjunctional electrical responses and, in particular, excitatory junction potentials (EJP). 2. Reserpine (5 mg/kg, i.p.) caused almost total depletion (below detection levels) of the NA content of the vas deferens. However, spontaneous and evoked excitatory junction potentials (SEJP, EJP) and currents (SEJC, EJC) could still be recorded in NA-depleted tissues. The amplitude distribution of SEJC in control and reserpinized tissues was similar. 3. Facilitation of EJP and EJC was markedly slowed in reserpinized tissues, EJP taking 50-60 pulses to facilitate fully. However the amplitudes of fully facilitated EJP were comparable to EJP recorded in control tissues. 4. EJPs in reserpinized vasa deferentia were unaffected by the NA synthesis inhibitor, alpha methyl tyrosine, but were abolished in the presence of the stable ATP analogue alpha,beta-methylene ATP which desensitizes postjunctional P2-purinoceptors. 5. Local application of ATP, but not NA, mimicked the EJP. These results indicate that EJP are mediated by a non-noradrenergic neurotransmitter possibly ATP. PMID- 2548782 TI - The study of tachykinin receptors. AB - 1. Three high affinity receptors (NK1, NK2 and NK3) recognizing the C-terminal end of tachykinins have been described, with probable endogenous ligands substance P (SP-), neurokinin A and neurokinin B, respectively. 2. Other receptors recognizing the N-terminal regions of SP exist in adrenal medulla and central nervous system (CNS) with a low affinity receptor on mast cells. 3. NK1 receptors are associated with blood vessels, all types of smooth muscle, enteric ganglia and glands. NK2 receptors are found on gastrointestinal and urinary smooth muscle, but few peripheral NK3 receptors have been described. In contrast, NK1, NK3 but not NK2 receptors are widely distributed in the CNS. 4. Problems associated with the study of tachykinin receptors include low selectivity of endogenous tachykinins, poor antagonists and susceptibility of peptides to enzymatic degradation. PMID- 2548781 TI - Opiate receptors: ligands and methods of study. AB - 1. An introduction to the current knowledge of the mu-, kappa- and delta-opioid receptors will be given. 2. The many problems associated with opiate ligands with respect to selectivity, potency, slow receptor kinetics, instability and poor access to the central nervous system will be discussed. 3. Current systems for analysis of the receptor profile of opiate ligand activity will be given. 4. The properties of opioid ligands will be discussed in the context of the identified problems. PMID- 2548783 TI - Immune dysfunction in hypophosphatemic vitamin D-resistant rickets: immunoregulatory reaction of 1 alpha(OH) vitamin D3. AB - We investigated immunologic function in six cases with hypophosphatemic vitamin D resistant rickets (VDRR) before and after treatment with 1 alpha hydroxycholecalciferol (1 alpha(OH) vitamin D3). All cases suffered frequent episodes of infection, which tended to be more severe in the older patients. OKT9 , OKT10-, and OKM1-positive cells and adenosine deaminase (ADA) were significantly increased, whereas numbers and activity of natural killer (NK) cells were lower than normal before treatment. After administration of 1 alpha(OH) vitamin D3, however, the susceptibility to infection apparently decreased, and NK cell number and activity increased in all patients. ADA was also significantly decreased and remained in the normal range after treatment. These results suggest that vitamin D plays a role in the impaired immunoregulatory functions of NK cells in VDRR. Furthermore, ADA may be one parameter reflecting this immunologic impairment. PMID- 2548784 TI - Physiological significance of increased levels of endogenous atrial natriuretic factor in human acute renal failure. AB - In twelve patients with acute renal failure, mean plasma levels of atrial natriuretic factor (ANF) and of its second messenger cGMP were found elevated at the early phase of the disease, but tended to return towards normal values at recovery. Variations of plasma ANF and cGMP were correlated significantly (p less than 0.05) with those of total blood volume. At the early phase of the disease, plasma ANF was also correlated with the excreted fraction of filtered sodium (FENa) (r = 0.95). Moreover, plasma ANF and FENa peaked concomitantly at the onset of the diuretic phase in the five patients who were not treated by diuretics or dialysis and were studied sequentially during the course of the disease. It is suggested that enhanced plasma ANF levels might reflect one of the mechanisms of adaptation controlling body fluid balance in acute renal failure. PMID- 2548785 TI - Neurological complications of infection with human immunodeficiency virus type 1. A review of literature and 241 cases. PMID- 2548786 TI - Place of the extra-intra-cranial-bypass in aneurysm-surgery. AB - Before the use of the EC/IC-bypass, despite several tests to predict complications, ligation of the i.c.a. (e.g. in treatment of an unclippable aneurysm) offered a significant risk of cerebral ischemia, either acutely or on long term. In this article the hemodynamic background of the EC/IC-bypass as well as its probable effect on local cerebral blood flow are discussed. Next, a series of 26 patients is presented in which ligation of the i.c.a or the m.c.a. was considered. To prevent cerebral ischemia, first an EC/IC-bypass was established. After all, in 15 patients ligation of the i.c.a. was necessary. Beside one major complication the other complications after ligation were only minor and temporary. PMID- 2548787 TI - Unruptured aneurysms in patients with transient ischemic attack or reversible ischemic neurological deficit. Report of eight cases. AB - Thirty-five patients with unruptured aneurysms were treated between 1981 and 1987. Eight of them had either transient ischemic attacks or reversible ischemic neurological deficits as their presenting symptoms. Six of the eight patients underwent direct aneurysm surgery. All eight patients are well, and have had no recurrent attack during the follow-up period ranging from 2 to 7 years. The feasibility of surgical treatment for such aneurysms associated with reversible ischemic symptoms is discussed. PMID- 2548788 TI - Subcortical arteriosclerotic encephalopathy--a controlled psychometric study. AB - Eleven patients with subcortical arteriosclerotic encephalopathy (SAE) were assessed psychometrically, and the results compared with control subjects without neurological disorders and patients with a single cerebral vascular lesion (CVL). The groups were matched for sex, age, years of education, and in addition for the CVL group, the nature, site and size of the discrete vascular lesion. The SAE patients had mild to moderate impairment in intellectual, memory, receptive language and constructional functions, but individual performance ranged from near normality to severe impairment. The differences between the performance of SAE patients and the controls were significant in the majority of tests. The SAE patients had lower scores than CVL patients on all tests, but in many of these the difference was not significant. PMID- 2548789 TI - Doppler and duplex sonography of the cervical arteries and correlations with other examinations. A field study in a population over forty years. AB - Of the population of a small Tyrolean village, 185 (56%) of the 329 inhabitants over 40 years were investigated by means of Doppler and duplex sonography, electroencephalography (EEG), electrocardiography (ECG), and neurological examination. Four subjects (2%) previously had a transitory ischemic attack (TIA) or stroke in the carotid territory. Sonographically detectable abnormalities in one or more extracranial arteries were present in 42 (23%) persons. Of the 14 subjects with more than slight abnormalities in the common or internal carotid artery two were symptomatic. Two additional cases with TIA or stroke did not show relevant lesions on sonographic examination. The presence of narrowing in the extracranial artery was not related to risk factors (hypertension, smoking, obesity) or abnormalities on ECG and EEG. This study shows 1) that the clinical relevance of ultrasound screening of the carotid arteries in an average population is 15%; 2) that significantly more patients with TIAs or strokes are found in the group with more severe sonographic findings (p = 0.001) than in the group with normal ultrasound results. PMID- 2548791 TI - Autoimmune thyroiditis and myopathy. Reversibility of myopathic alterations under thyroxine therapy. AB - A female patient with Hashimoto's autoimmune thyroiditis and myopathic symptoms is reported. Besides a myopathic pattern, widespread spontaneous pathological activity was found in the affected musculature. Under thyroxin therapy, both the clinical picture and the electromyographic findings normalized completely within a few weeks. PMID- 2548790 TI - Steroid-induced complete remission in a case of primary cerebral non-Hodgkin's lymphoma. AB - A case of a primary non-Hodgkin's lymphoma of the CNS is reported in which a complete and stable disappearance of an enhancing mass lesion on the CT scan after treatment with dexamethasone was followed by the occurrence of a high-grade malignant lymphoma in the opposite hemisphere 18 months later. The long-lasting and stable remission suggests a direct oncolytic effect of corticosteroids on lymphoma cells. Furthermore, this case illustrates the usefulness of repeated CT examinations in patients with CNS lymphoma and steroid treatment. PMID- 2548792 TI - Sulindac-induced aseptic meningitis in mixed connective tissue disease. AB - A 21-year-old female with mixed connective tissue disease (MCTD) experienced nausea, headache, consciousness disturbance, nuchal rigidity, and a temperature of 38.5 less than or equal to C three days after the intake of sulindac (300 mg/day). Cerebrospinal fluid analysis revealed an opening pressure of 310 mm of water, a predominantly lymphocytic pleocytosis, and elevated protein content of 89 mg/dl. After discontinuing sulindac, the aseptic meningitis improved in five days. In the acute stage, CT scan disclosed contrast enhancement in the cerebral hemispheres, which suggests that hypersensitivity may be involved in the pathogenesis of nonsteroidal antiinflammatory drug (NSAID) induced aseptic meningitis. PMID- 2548793 TI - Pelizaeus-Merzbacher disease with thiamine deficiency or Leigh disease with extensive involvement of white matter? Case report. AB - We report on a case of a 30-month-old child who presented with a clinical syndrome compatible with leucodystrophy and in whom neuropathological features of both Pelizaeus-Merzbacher disease and subacute necrotizing encephalopathy were shown. The significance of the neuropathological findings is discussed in the light of a possible coexistence of both diseases which has not previously been reported. PMID- 2548794 TI - Multiple brain abscesses secondary to bronchiectasis. A case of 34 discrete abscesses in one brain. AB - Multiple brain abscesses are a rare complication which may occur in a number of conditions including intra-thoracic sepsis. Computerized tomography has had an advantageous impact on the management of this condition by facilitating earlier diagnosis. However, the treatment modalities are inadequate and mortality remains high. A case of multiple brain abscesses in a patient with bronchiectasis is reported which exemplifies this often fatal condition. The management of multiple brain abscesses is discussed. PMID- 2548795 TI - Ossified epidural haematoma. Report of two cases. AB - Two patients of ossified epidural haematoma (EDH) following ventriculoperitoneal shunt for hydrocephalus are reported. Pathogenesis of an ossified EDH is discussed with special emphasis on post-shunt chronic EDH. PMID- 2548796 TI - Technetium-99m pertechnetate and gallium-67 imaging in salivary gland disease. AB - Thirty-two patients with salivary gland tumors or sialadenitis were studied with Tc-99m pertechnetate and Ga-67 imaging and, in some instances, sialography. The diagnostic algorithm presented allows the correct categorization of the salivary gland pathology in the vast majority of patients. The patients were studied serially with Tc-99m pertechnetate, Ga-67 and in certain situations sialography (or CT-sialography). Use of the algorithm can distinguish benign salivary tumors from malignant tumors and malignant tumors from inflammatory disease. The limitations and pitfalls of interpretation are discussed. PMID- 2548798 TI - Examples of complementary roles of abdominal CT and radiocolloid liver imaging. AB - Two cases are reported in which well-defined lesions of the liver were seen in radiocolloid liver imagings but not in the concurrent abdominal CT images. The results of autopsy confirmed the diagnosis of hepatoma in one and metastatic disease in the other. The third case, referred for a Tc-99m SC liver-spleen image because of a suboptimal abdominal CT owing to artifact, showed a normal liver. In certain clinical situations, radiocolloid imaging provides a complementary role to CT in the detection or confirmation of hepatic lesions and, in occasional patients, is superior to CT. PMID- 2548797 TI - Comparison of delayed imaging with Tc-99m PMT and Tc-99m DEIDA for visualization of hepatoma. AB - Delayed imaging was performed after the intravenous administration of Tc-99m DEIDA and Tc-99m PMT in 18 patients with hepatocellular carcinoma. Using Tc-99m DEIDA imaging, sharp uptake by liver tumors was observed in four patients (22%), but the uptake was similar to that of the surrounding normal liver in eight patients (44%). Using Tc-99m PMT imaging, the uptake by the tumor was notable in ten patients (56%) and normal in two (11%). Tc-99m PMT and Tc-99m DEIDA were both concentrated in hepatocellular carcinomas, but the former showed intense uptakes more frequently, and thus is suggested to be useful in the diagnosis of hepatocellular carcinoma. PMID- 2548799 TI - Tc-99m pyrophosphate localization in chest wall muscles after bench pressing. PMID- 2548800 TI - Serum digoxin-like substances in pregnancy-induced hypertension. AB - 1. Endogenous digoxin-like immunoreactivity (EDLI) was measured in the serum of 85 normotensive pregnant (NTP) women and 77 women with pregnancy-induced hypertension (PIH) by a radioimmunoassay (New England Nuclear). All women were in the third trimester. 2. EDLI, which was undetectable in serum from non-pregnant women, was present in NTP and PIH and was significantly higher in PIH. EDLI correlated with gestational age in NTP, but not in PIH. 3. Ouabain-sensitive Na+ transport was estimated in normal peripheral blood leucocytes after incubation with sera from 50 NTP and 42 PIH women. Significant inhibition of active Na+ transport occurred only with the serum of hypertensive patients without proteinuria. 4. EDLI did not correlate with the effect of the sera on active Na+ transport. The radioimmunoassay therefore provides a poor index of Na+ transport inhibitory activity in PIH. PMID- 2548801 TI - Lipoxin A4 and lipoxin B4 inhibit chemotactic responses of human neutrophils stimulated by leukotriene B4 and N-formyl-L-methionyl-L-leucyl-L-phenylalanine. AB - 1. Lipoxin A4 (LXA4) and lipoxin B4 (LXB4) have been evaluated for their capacities to modulate neutrophil (PMN) migration and endothelial cell adherence using compounds prepared by total chemical synthesis. 2. Increased PMN migration was seen with concentrations of LXA4 from 10(-9) mol/l to 10(-7) mol/l. LXA4 was 100-fold less potent than leukotriene B4 (LTB4) and it elicited only one-half of the maximal response of LTB4. 3. The (5S,6S,15S)-isomer of LXA4 induced only a weak migratory response and LXB4 was inactive, suggesting that the activity of LXA4 was stereospecific. 4. Modified chequerboard analysis indicated that LXA4 was a chemokinetic agent. 5. Preincubation of PMN with increasing concentrations of LXA4 induced a very similar dose- and time-dependent inhibition of the subsequent response to 10(-7) mol/l LTB4 or 10(-7) mol/l N-formyl-L-methionyl-L leucyl-L-phenylalanine (FMLP). The inhibition was observed at 10(-10) mol/l LXA4; the concentration which produced 50% inhibition was 10(-8) mol/l and 100% inhibition of PMN locomotion occurred at 10(-6) mol/l LXA4. 6. The (5S,6S,15S) isomers of LXA4 and LXB4 were 5- and 100-fold less potent than LXA4, respectively, in suppressing LTB4- or FMLP-induced PMN migration. 7. Preincubation of PMN with LXA4 led to a suppression of calcium mobilization, as assessed by Quin2-AM fluorescence, when the cells were subsequently stimulated under optimal conditions by LTB4 or FMLP. 8. These results suggest that the inhibitory activity of lipoxins may be related to the capacity of these molecules to regulate calcium ion mobilization. PMID- 2548802 TI - Recommended methods for an additional red cell enzyme (pyrimidine 5' nucleotidase) assay and the determination of red cell adenosine-5'-triphosphate, 2,3-diphosphoglycerate and reduced glutathione. International Committee for Standardization in Haematology. PMID- 2548803 TI - Cardiac hypertrophy: from fetal to fatal? PMID- 2548804 TI - [Action of evening primrose oil on cardiovascular risk factors in insulin dependent diabetics]. AB - In an open study, the authors compared two groups of insulin-dependent diabetics matched for age and metabolic control, one of which was given a linoleic-gamma linolenic acid mixture (3 g daily), the other served as control. The effect, attributed to gamma-linolenic acid only, was evaluated as explained in the text and is shown in the table. At the end of two months no change was found in the control group while favorable changes of HDL-cholesterol and platelet adhesiveness were observed in the experimental group. PMID- 2548805 TI - Insulin-like growth factor I receptor analysis of satellite cell-derived myotube membranes established from two lines of Targhee rams selected for growth rate. AB - Ovine-derived fibroblasts were used to validate an insulin-like growth factor I (IGF-I) membrane-receptor binding assay system. Competitive binding using fibroblasts revealed that half-maximal inhibition of 125I-IGF-I binding by IGF-I was 2.3 nM. SDS-polyacrylamide gel electrophoresis analysis of specific protein associated 125I-IGF-I was consistent with the migration of 125I-IGF-I-labeled Type I IGF receptor alpha-subunits at Mr 133,000 daltons. Further, the efficiency of two cell solubilization methods was examined and time-dependent binding equilibrium was determined for the membrane assay system. Satellite cell-derived myotubes were subsequently isolated from primary satellite cell cultures established from the semimembranosus muscles of high and low efficiency-of-gain (EOG) Targhee rams, and IGF-I receptor dynamics were measured. A membrane competitive binding study revealed that half-maximal inhibition of 125I-IGF-I binding was achieved by 1-ng IGF-I for low, and 10-ng IGF-I for high, EOG myotube membrane preparations. Kd values were similar between the high EOG (4.78 nM) and low EOG (2.95 nM) groups; however, receptor concentrations (Bmax) appeared to differ between groups. High EOG membrane receptor Bmax was 3.88 pmole/micrograms protein (19.87 pmole/micrograms DNA), whereas low EOG membrane receptor Bmax was 1.22 pmole/micrograms protein (9.28 pmole/micrograms DNA). These preliminary findings support the hypothesis that genetic selection for EOG results in altered satellite cell responsiveness to IGF-I. PMID- 2548806 TI - Allergic contact dermatitis from the surfactant in Hibitane. PMID- 2548808 TI - Neurologic abnormalities of uremia. PMID- 2548807 TI - Gestodene: a novel synthetic progestin--characterization of binding to receptor and serum proteins. AB - Gestodene, 17 alpha-ethinyl-13-ethyl-17 beta-hydroxy-4,15-gonadien-3-one, is a new orally active progestational agent, which is available for clinical use in oral contraceptives. The aim of the present study is to make a broad characterization of gestodene at the receptor level and to discuss the results in comparison to those of established progestogens. Kinetic studies of 3H-gestodene uptake show a rapid increase in the amount of specific binding during the first three hours. After saturation, the amount of specifically bound 3H-gestodene remained almost constant up to 24 hours at 4 degrees C. The dissociation of 3H gestodene from the cytoplasmic myometrial progestone receptor, measured by displacement of labeled steroid with dextran-coated charcoal treatment at 4 degrees C at various times, showed a biphasic or two-component first order dissociation curve. As anticipated, sucrose gradient centrifugation analysis of the 3H-gestodene-labeled cytosol of human myometrial tissue showed that the gestodene binding components sedimented in the 4S and 8S region. A 200-fold molar excess of nonradioactive gestodene reduced only the 8S binding of 3H-gestodene. 4S binding of 3H-gestodene was not reduced, which indicate the existence of a second high capacity binding component. In biological test systems, such as the Clauberg test or Kaufmann test, gestodene has proved to be a very effective progestogen. Among nortestosterone derivatives it is one of the most potent and resembles progesterone biologically in its progestogenic effects. This biologically identical gestagenic activity of gestodene and progesterone is reflected by a very similar behavior in vitro in terms of binding to progesterone receptors of human uterus cytosol. Furthermore, competitive studies indicated that gestodene like other synthetic progestagens also displays some affinity for androgen and glucocorticoid receptors but no measurable affinity for the estrogen receptor. Remarkable is the high binding affinity of gestodene to the binding sites of the mineralocorticoid receptor of rat kidney with a RBA value of 350% compared to aldosterone. PMID- 2548809 TI - Molecular confirmation of an abortigenic strain of equine herpesvirus 1 (subtype 1) in a pregnant mare study. AB - Four pregnant mares were inoculated intranasally and/or intravenously with equine herpesvirus 1 (EHV-1), subtype 1 during the third trimester of gestation. One mare aborted on postinfection day 15, one mare delivered a sick, weak full term foal, and two mares delivered healthy, full term foals. EHV-1, subtype 1 was isolated from several tissues of the aborted fetus and from the thymus of the sick foal. DNA restriction endonuclease patterns of the recovered EHV-1 viruses were identical to those of the EHV-1 challenge strain, documenting the origin of the abortigenic viruses. PMID- 2548810 TI - Leukocytic oxygen activation and microbicidal oxidative toxins. AB - Following a brief introduction of cellular response to stimulation comprising leukocyte activation, three major areas are discussed: (1) the neutrophil oxidase; (2) myeloperoxidase (MPO)-dependent oxidative microbicidal reactions; and (3) MPO-independent oxidative reactions. Topics included in section (A) are current views on the activation mechanism, redox composition, structural and topographic organization of the oxidase, and its respiratory products. In section (B), emphasis is placed on recent research on cidal mechanisms of HOCl, including the oxidative biochemistry of active chlorine compounds, identification of sites of lesions in bacteria, and attendant metabolic consequences. In section (C), we review the (bio)chemistry of H2O2 and .OH microbicidal reactions, with particular attention being given to addressing the controversial issue of probe methods to identify .OH radical and critical assessment of the recent proposal that MPO independent killing arises from site-specific metal-catalyzed Fenton-type chemistry. PMID- 2548811 TI - Microbial amylolytic enzymes. AB - Starch-degrading, amylolytic enzymes are widely distributed among microbes. Several activities are required to hydrolyze starch to its glucose units. These enzymes include alpha-amylase, beta-amylase, glucoamylase, alpha-glucosidase, pullulan-degrading enzymes, exoacting enzymes yielding alpha-type endproducts, and cyclodextrin glycosyltransferase. Properties of these enzymes vary and are somewhat linked to the environmental circumstances of the producing organisms. Features of the enzymes, their action patterns, physicochemical properties, occurrence, genetics, and results obtained from cloning of the genes are described. Among all the amylolytic enzymes, the genetics of alpha-amylase in Bacillus subtilis are best known. Alpha-Amylase production in B. subtilis is regulated by several genetic elements, many of which have synergistic effects. Genes encoding enzymes from all the amylolytic enzyme groups dealt with here have been cloned, and the sequences have been found to contain some highly conserved regions thought to be essential for their action and/or structure. Glucoamylase appears usually in several forms, which seem to be the results of a variety of mechanisms, including heterogeneous glycosylation, limited proteolysis, multiple modes of mRNA splicing, and the presence of several structural genes. PMID- 2548812 TI - Regional treatment of hepatic metastases and hepatocellular carcinoma. AB - Hepatic metastases represent a common site of dissemination for a number of primary malignancies related in part to the dual blood supply, large blood flow, and receptive environment of the hepatic parenchyma. Although this review focuses on regional therapy, we have included sections on systemic therapy to better interpret the results with intrahepatic therapy. We will also discuss the efficiency of hepatic arterial ligation, embolization, and radiotherapy of hepatic metastases. Primary gastrointestinal neoplasms are particularly prone to produce hepatic metastases. Because colorectal carcinoma metastasizes to the liver in up to 70% of patients with advanced disease, the treatment of hepatic metastases is a relevant topic. We will discuss the systemic and regional therapy of colorectal, gastric, and gallbladder cancers. Breast carcinoma and malignant melanoma frequently metastasize to the liver, and we have described systemic and regional treatments of these diseases. Because sarcomas are often treated by regional therapy, we have included a section on the treatment of hepatic sarcomas. Neuroendocrine tumors (carcinoid and islet cell), although often slow growing, frequently metastasize to the liver and then cause symptomatic problems. Much of the work done with embolization and hepatic ligation in the treatment of hepatic metastases has been performed in neuroendocrine tumors, and these studies, as well as the systemic and regional chemotherapy of hepatic metastases, will be described. The last section concerns the treatment of hepatocellular carcinoma. We have outlined the staging systems used. We then detail the results of systemic and intrahepatic therapy, embolization, and hepatic ligation in the treatment of hepatocellular carcinoma. Because hepatic metastases are a frequent problem, many patients are available for clinical investigation. It is hoped that newer strategies for the treatment of liver metastases will lead to higher response rates and perhaps control of local disease. These therapeutic approaches may also give us leads to the treatment of systemic disease. PMID- 2548813 TI - The pituitary-specific form of rat c-erbA is a biologically active thyroid hormone receptor. PMID- 2548814 TI - Herpes simplex virus latency analyzed by in situ hybridization. PMID- 2548815 TI - Nedocromil sodium and placebo in the treatment of bronchial asthma. A multicenter, double-blind, parallel-group comparison. AB - The efficacy of nedocromil sodium (4 mg twice daily by inhalation) in treating bronchial asthma was assessed by double-blind, placebo-controlled group comparison in 69 adults from three centers. The patients (34 active, 35 placebo) had a history of bronchial asthma with at least 15 percent reversibility. Inhaled corticosteroids, used by 22 and 24 subjects in the active and placebo groups respectively, were discontinued before the study, in which a two-week baseline was followed by six weeks of treatment. Two-weekly clinic assessments of lung function, symptoms and final opinions of treatment were significantly (p less than 0.05 p less than 0.001) in favor of nedocromil sodium. Daily diary cards showed a similar trend with significant drug effects seen after the third week. Blood and urine samples showed no abnormalities and the majority of patient withdrawals (five from nedocromil sodium and six from placebo treatment) were due to worsening asthma. Overall, we found nedocromil sodium to be well tolerated and effective in the management of bronchial asthma. PMID- 2548816 TI - Traumatic sternal segment dislocation in a child. AB - We present an extremely rare case of traumatic dislocation of a sternal body segment in a child. We treated this sternal segment dislocation by means of open reduction using a newly developed pin made from poly-L-lactide containing 10 percent by weight hydroxyapatite. It is not necessary to remove this type of pin later because it is degradable and absorbed within the body within about one year after implantation. A two-year follow-up of this case revealed a good clinical result. PMID- 2548817 TI - Comparison of in vitro activity of daptomycin, vancomycin and fluoroquinolones in broth and serum against Staphylococcus epidermidis as determined by time-kill kinetics. AB - We compared the activity of daptomycin, vancomycin and fluoroquinolones against 46 blood isolates of Staphylococcus epidermidis. Daptomycin was more rapidly bactericidal than vancomycin and demonstrated less of an inoculum effect than the quinolones. Time-kill kinetic studies demonstrated no significant differences in survival in broth as compared to 50% human serum-supplemented broth. Daptomycin and the quinolones are suitable for study in clinical trials of efficacy as therapeutic alternatives to vancomycin for S. epidermidis infections. PMID- 2548818 TI - Treatment of experimental zygomycosis in guinea pigs with azoles and with amphotericin B. AB - Nonpredisposed Albino guinea pigs were infected intravenously with Rhizopus microsporus var. rhizopodiformis or with Rhizopus oryzae. Both strains were highly pathogenic. They killed all control animals between days 4 and 7 and between days 5 and 9 after infection, respectively. All animals presented invasion of almost all internal organs and skin eruptions developing into ulcers. Oral treatment with ketoconazole, itraconazole, fluconazole or saperconazole was inefficacious. Parenteral treatment with amphotericin B prolonged survival and was life-saving in 9 out of 12 guinea pigs infected with Rh. microsporus var. rhizopodiformis and in 5 out of 12 infected with Rh. oryzae. More active therapy is needed. PMID- 2548819 TI - Using transposon Tn5 insertions to sequence bacteriophage T4 gene 11. AB - A simple and rapid method of creating an overlapping set of deletions in cloned DNA in preparation for sequencing has been developed. The method is based on a positive selection for Tn5 transposition into the cloned DNA fragment on a high copy-number filamid, resolution of potential filamid dimers by filamentous phage infection, and the use of Tn5 both as a "portable" restriction enzyme site for in vitro DNA deletion and a "portable" sequencing primer binding site to initiate DNA sequencing reactions using a custom primer complementary to the outside ends of IS50. This new method has been utilized to sequence bacteriophage T4 gene 11, encoding the T4 baseplate protein gp11. The coding sequence of gene 11 is 657 bp in length, and predicts a primary structure of 219 amino acids that agrees well with the biochemical data previously obtained. DNA sequence around gene 11 suggests that the expression of genes 10, 11, and 12 of phage T4 are translationally coupled. Plasmids carrying deletions generated using this method have been used to map genetically five amber alleles of gene 11. These amber alleles were sequenced to confirm the proposed reading frame. The five amber alleles actually represent two different mutational changes at either codon 206 or 207, changing these adjacent glutamine codons to amber. The position of these amber alleles lends support to earlier studies identifying the carboxyl terminus of gp11 as essential in the interaction of P11 with baseplate protein P10 (Plishker and Berget, 1984). PMID- 2548820 TI - A rapid procedure to identify newborn transgenic mice. AB - We have developed a rapid procedure to identify newborn transgenic mice containing foreign genetic material in their genome. The protocol involves collagenase digestion of a small amount of tail tissue which can be taken very early after birth, phenol and chloroform extraction, polymerase chain reaction, and polyacrylamide gel electrophoresis. The entire procedure, from tissue biopsy to final results, can be completed in 1 day. PMID- 2548821 TI - [Left ventricle-right atrium communication diagnosed by Doppler and contrast echocardiography]. PMID- 2548822 TI - [Effects of PGE2 on experimental atherosclerosis]. PMID- 2548823 TI - [Post-menopausal virilism]. PMID- 2548824 TI - [A new herpes virus]. PMID- 2548825 TI - [Chronic fatigue symptoms--a new syndrome associated with persistent exhaustion]. PMID- 2548826 TI - Electrophysiological findings in crutch palsy. AB - Detailed electromyographic studies in two patients suffering from crutch palsy revealed conduction blocks along the nerves to the involved upper limbs localized to the axilla. Rapid reversal of the blocks was documented in one patient. Electromyography is of value in delineating such lesions and helping in the formulation of prognosis. PMID- 2548827 TI - Identification of motor conduction block despite desynchronisation. A method. AB - Identification of incomplete motor conduction block (CB) by the usual single stimulation method is problematic when impaired conduction velocity of the axons spared by the block exists at the site of the presumed CB, leading to desynchronisation. Indeed, the area of the compound muscle action potential (MAP) evoked by stimulation applied above this site may be reduced by the phase cancellation related to desynchronisation. A triple stimulation method with double collisions allows virtual displacement of the CB distally to the site where desynchronisation occurs. A first stimulus applied on the nerve above the site of the CB is followed--after a delay--by a stimulation applied distally on the nerve where a collision occurs. This collision, however, does not concern the antidromic wave of the blocked axons which travels towards the block. A third stimulus is then applied--after a second delay--below the site of the CB, where a second collision occurs in the blocked axons. Thus, the third stimulus evokes a MAP that concerns only the unblocked axons, as would the stimulus applied above the site of the CB. In this manner, desynchronisation occurring at the CB site is avoided. The method was developed on simulated CBs of the ulnar nerve at the elbow of 33 normal subjects and compared with the usual single stimulation method. Subsequently, it was evaluated on 8 pathological CBs of the ulnar nerve at the elbow and of the peroneal nerve at the fibular head. The method detects CBs of 5% or more despite the occurrence of desynchronisation. PMID- 2548828 TI - Studies on uptake and utilization of high density lipoprotein cholesterol for steroid hormone synthesis in the isolated adrenocortical cells. PMID- 2548829 TI - Quantal charge redistributions accompanying the structural transitions of sodium channels. AB - Asymmetric displacement currents, Ig, associated with the gating of nerve sodium channels have been recorded in cell-attached macropatches of Xenopus laevis oocytes injected with exogenous mRNA coding for rat-brain-II sodium channels. The Ig properties were found to be similar to those of gating currents previously observed in native nerve preparations. Ig fluctuations were measured in order to ascertain the discreteness of the conformational changes which precede the channel opening. The auto-correlation of the fluctuations is consistent with a shot-like character of the elementary Ig contributions. The variance of the fluctuations indicates that most of the gating-charge movement that accompanies the activation of a single sodium channel occurs in 2 to 3 brief packets, each carrying an equivalent of about 2.3 electron charges. PMID- 2548830 TI - Stimulation of arachidonic acid metabolism in silica-exposed alveolar macrophages. AB - The molecular events involved in both the initiation and development of silicosis are at present poorly defined, although mediators released from macrophages exposed to silica particles are believed to play a role. We have investigated the in vitro production of arachidonic acid (AA) metabolites in adherent bovine alveolar macrophages (BAM) incubated with crystalline silica. BAM were prelabeled with 3H-AA and incubated with 0.5-5.0 mg silica. Lipid metabolites released into the culture medium were analyzed by high-performance liquid chromatography. Simultaneously, lactate dehydrogenase (LDH) was assayed to provide an indication of cell injury. No 5-lipoxygenase metabolites were detected at the lowest silica dose tested (0.5 mg/well), but 5-hydroxyeicosatetraenoic acid (5-HETE) was the major AA metabolite detected between 1.5 and 5.0 mg of silica. A fivefold increase in the production of leukotriene B4 (LTB4) and its two nonenzymatic diastereomers (Isomers I and II) was observed as the silica concentration was increased from 1.0 to 5.0 mg. In contrast, the release of cyclooxygenase products declined with increasing concentrations of silica. LDH release increased in a linear, dose-dependent fashion in the range of silica doses used. The kinetics of eicosanoid release was investigated over a 3-h interval and LDH release was assayed for each time point. Within 15 min following silica addition, a shift to the production of 5-lipoxygenase metabolites was observed, accompanied by a reduction in cyclooxygenase products. This rapid alteration in AA metabolism preceded cell injury as measured by LDH release. These results demonstrate that silica is a powerful stimulator of arachidonic acid metabolism in BAM. Moreover, silica selectively stimulates the 5-lipoxygenase pathway as the dose of silica increases. Our results suggest that dysfunction in arachidonate metabolism could contribute to the pathogenesis of silicosis. PMID- 2548831 TI - Role of cyclic AMP in proliferation of lung tissue in organ culture. AB - The role of cyclic AMP in cell proliferation and division has been the subject of study by a number of investigators in the past 30 years, but the argument of whether cyclic AMP is a negative or a positive regulator has not been settled. We studied the effect of cyclic AMP on proliferation of normal and postpneumonectomized lung tissues in young adult rats by measuring the incorporation of tritiated thymidine into lung DNA in organ culture. In normal lung tissues the incorporation of [3H]thymidine was increased by exogenous dibutyryl cyclic AMP, or by isoproterenol or forskolin to stimulate adenylate cyclase, or by caffeine, which inhibits cAMP phosphodiesterase. The effect of isoproterenol, but not forskolin, was abolished by the beta-adrenergic blocking agent propranolol. The effect of caffeine on [3H]thymidine incorporation was further enhanced in normal lung tissues in the presence of dibutyryl cyclic AMP and in postpneumonectomized lung tissues. Imidazole, a cAMP phosphodiesterase stimulator, also increased [3H]thymidine incorporation in culture, but the effect was not magnified in the presence of exogenous dibutyryl cyclic AMP, nor in postpneumonectomized lung tissues. The data suggest that cyclic AMP acts as a positive regulator in proliferation of lung tissues. PMID- 2548832 TI - Effects of centrophenoxine on cholinephosphotransferase activity in maternal and fetal guinea pig lung. AB - Cholinephosphotransferase activities of guinea pig lung mitochondria and microsomes are inhibited by centrophenoxine and one of its metabolites, p chlorophenoxyacetate. 2-Dimethylaminoethanol, the second metabolite of centrophenoxine, has no inhibitory effect on the enzyme activity. The inhibition of enzyme activity by centrophenoxine is noncompetitive. Intravenous injection of centrophenoxine and p-chlorophenoxyacetate to pregnant animals causes inhibition of cholinephosphotransferase activity in adult lung but not in fetal lung. However, direct administration of centrophenoxine to the fetus after laparotomy causes inhibition of both subcellular enzyme activity in fetal lung. It is suggested that the drug injected to the pregnant animals does not reach the fetal lung or is metabolized. Furthermore, while centrophenoxine injection does not change the total phosphatidylcholine content of adult lung, the acyl group composition of phosphatidylcholine was modulated. PMID- 2548833 TI - Characterization of scavenger receptor activity in resident human lung macrophages. AB - We characterized scavenger receptor pathways of human alveolar macrophages and cultured monocytes using radiolabeled maleylated bovine serum albumin (MAL-BSA) and acetylated low-density lipoprotein (Ac-LDL) as ligands. Human alveolar macrophages and cultured human monocytes degraded both MAL-BSA and Ac-LDL. Both ligands were bound and degraded in a specific and saturable fashion. Specificity of degradation was tested using excess MAL-BSA and Ac-LDL, polyanionic compounds, and alpha-casein as inhibitors. Alveolar macrophages utilized the classical scavenger receptor pathway to degrade MAL-BSA and Ac-LDL. In contrast, cultured monocytes utilized two receptor pathways to degrade MAL-BSA: the classical scavenger receptor pathway and a secondary alpha-casein-inhibitable pathway. These results demonstrate differences in the activities of receptor systems in cultured monocytes compared to alveolar macrophages. PMID- 2548834 TI - Search for drugs that may reduce the load of neutrophil azurophilic granule enzymes in the lungs of patients with emphysema. AB - Neutrophil elastase and myeloperoxidase probably play an important role in the development of pulmonary emphysema. We have analyzed drugs from the major classes of agents that alter neutrophil function to determine if there are drugs in use today that can reduce the load of neutrophil elastase or myeloperoxidase in the lungs of smokers. Eleven representative drugs were tested for their ability to inhibit chemotaxis and degranulation. None of the drugs inhibited chemotaxis in a dose-response fashion at concentrations achievable in human plasma. Sulfinpyrazone, phenylbutazone, and auranofin completely inhibited the release of azurophilic granules (myeloperoxidase) and tertiary granules (beta-D glucuronidase) when formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP) was used as the stimulant, and inhibited azurophilic granule release by 69%, 19%, and 64% respectively, but not tertiary granule release when macrophage-conditioned media was used as the stimulus. In conclusion, none of the drugs tested are inhibitors of chemotaxis; however, three are excellent inhibitors of azurophilic granule enzyme release. Of these three, sulfinpyrazone, a drug that is not currently used clinically for its antiinflammatory effects, is the least toxic and should be considered as a potential drug to reduce the elastase and myeloperoxidase load in the lungs of smokers who are developing emphysema. PMID- 2548835 TI - Peroxidase activities in the hamster bronchoalveolar lining fluid: modifications induced by exposure to silica dust. AB - The modifications of peroxidase (Po) activity have been studied in bronchoalveolar lavage fluid (BALF) from hamsters exposed to silica dust. In silica-treated animals, the mean total BALF-Po activity was significantly increased compared to control animals. This increased activity was accompanied by an influx of polymorphonuclear neutrophils in airways. HPLC gel filtration of BALF from control animals separated 5 peaks with Po activity. They had an apparent molecular weight of 140, 110, 80, 57, and 42 kDa. In BALF from silica exposed animals, with the exception of the 57-kDa fraction, the same peaks were found. Additional fractions with an apparent molecular weight of greater than 200, 180, 92, 65, and 20 kDa were detected. All the fractions but those at 57 and 92 kDa were detectable in a whole-blood homogenate. Exposing hamsters to silica induced both quantitative modifications and a different pattern of BALF proteins having Po activity in the alveolar lining fluid. PMID- 2548836 TI - GABA in epilepsy: the pharmacologic basis. AB - GABA transmission plays a key role in controlling seizure activity. The precise nature of its effect depends on the particular location in the brain and the pathway involved. Animal studies have helped to define specific brain regions such as the substantia nigra and area tempestas that are critical in controlling seizure activity. Antiepileptic drugs such as vigabatrin [gamma vinyl GABA (GVG)], a drug rationally developed to treat resistant epilepsy, can enhance GABA transmission in these regions and may thereby afford seizure protection. PMID- 2548837 TI - Gel electrophoretic analysis of DNA branched junctions. AB - Gel electrophoresis has provided much of the detailed information we have about the properties of DNA junctions, stable branched molecules formed from oligonucleotide or polynucleotide strands. Here we review these applications, and present the results of an electrophoretic investigation of conformationally restricted junctions formed by covalently connecting two different pairs of strands in a junction with four arms. Native gel electrophoresis is employed to establish the formation and stoichiometry of the multistrand complexes. Ferguson analysis of native gel mobility shows that junctions have retardation coefficients that are distinct from those of linear DNA duplexes. Denaturing gel electrophoresis is the primary tool for characterizing junctions that have been covalently linked together to form both linear and macrocyclic oligomers of junctions (oligojunctions). Radioactively labelled strands enable one to monitor the progress of the ligation reaction: both linear and closed cyclic molecules result, and these can be distinguished by applying Ferguson analysis to denaturing gels. Combinations of exonuclease III, restriction enzymes and sequencing reactions have been applied to oligojunction molecules, and the results are all analyzed on denaturing gels. Junctions containing intramolecular "tethers" that restrict the conformation freedom of the complex comprise a new system for analyzing the conformations of branched molecules. In these tethered junctions, the ability of arms to move relative to each other is restricted substantially by covalently connecting pairs of arms in the original complex with short, flexible loops. The two tethers used here constrain the helical domains of the structure to be roughly parallel or anti-parallel. In this article, we use Ferguson analysis to compare two tethered junctions with an untethered junction. At high gel concentrations, the mobility of the untethered complex is found to be closer to that of the molecule tethered anti-parallel than to the one tethered parallel. Curvature in the Ferguson plots for all three of these junctions is detected over a range of compositions. At low gel concentrations, differences in electrophoretic mobility persist, suggesting that the untethered junction differs in charge as well as conformational freedom from the tethered analogs. We expect that studies of this kind will be able to define the conformational repertoire of junctions of different kinds, and to explore the effects of electrophoresis on these states. PMID- 2548838 TI - The migration anomaly of DNA fragments in polyacrylamide gels allows the detection of small sequence-specific DNA structure variations. AB - Curved DNA fragments have a reduced electrophoretic mobility in polyacrylamide gels. The retardation in gels is extremely sensitive to small structural variations which influence the DNA helix axis. This gel assay can also be used to detect very small structural variations in DNA sequences which are not curved: The noncurved sequences of interest can be combined with curved stretches in phase with the helix turn. Using such sequence constructions, even subtle influences on the DNA helix axis can be detected. Experiments of this kind allow the determination of a relative order of sequence-specific DNA twist and wedge angles. PMID- 2548839 TI - Studies of DNA-protein interactions by gel electrophoresis. AB - The use of gel electrophoresis in studies of nucleic acid-protein (especially DNA protein) interactions has yielded much qualitative and quantitative information about a variety of such systems. The reduction in mobility of complexes relative to free DNA allows isolation and characterization of the complexes as well as determination of thermodynamic and kinetic properties of the interactions. This article begins with a review of recent applications of the "gel retardation" assay, by way of introduction to experiments in two areas. In the first, a hypothesis is tested regarding whether a DNA molecule with sizable proteins bound very near to each end migrates through a polyacrylamide gel differently than does the corresponding complex having the proteins in the middle of the DNA fragment. The data show little mobility differences for these types of complexes, implying that both may move in a linear, "snakelike", manner through the gel. The experiments also provide results pertaining to questions of DNA bending caused by the binding of the E. coli catabolite activator protein (CAP) and RNA polymerase to the lactose promoter region. It appears that DNA bending by CAP at its wild type lac binding site is retained in complexes where RNA polymerase is bound simultaneously at the lac UV5 promoter. PMID- 2548840 TI - Quantitative footprinting analysis of drug-DNA interactions: Fe(III) methidium propyl-EDTA as a probe. AB - Quantitative footprinting studies involving a 139-base pair restriction fragment from pBR322 DNA, a lexitropsin ligand and two different DNA cleavage agents, the enzyme DNase I and the footprinting reagent Fe(III) methidium-propyl-EDTA (Fe MPE), are described. The autoradiographic data showed that the ligand, an analogue of netropsin possessing two N-methylimidazole groups, binds to four regions on the 139-mer which are rich in GC. Analysis of the data leading to individual binding constants for each of the four loading events on the 139-mer revealed that Fe-MPE and DNase I report the same binding constants for the lexitropsin bound to its interaction sequences. The fact that the data from both probes can be analyzed using a common model indicates that the DNA cleavage specificity of the probe and not its binding/cleavage mechanism is the important factor in reporting of site loading information in the footprinting experiment. The study also showed that under certain conditions it is possible to gain information on the density of ligand binding sites on carrier DNA by monitoring site loading events on the labeled fragment. PMID- 2548841 TI - An ice nucleation reporter gene system: identification of inducible pathogenicity genes in Pseudomonas syringae pv. phaseolicola. AB - We have constructed derivatives of the transposon Tn3 that allow an ice nucleation gene (inaZ) to be used as 'reporter' of the transcriptional activity of genes into which it is inserted. In these derivatives (Tn3-Ice and Tn3-Spice), the lacZYA sequences of transposon Tn3-HoHo1 were replaced with inaZ lacking its native promoter. The ice nucleation activity of virB::inaZ fusions in the correct transcriptional orientation was inducible by acetosyringone, a plant metabolite which activates the vir operon of Agrobacterium tumefaciens Ti plasmids, while fusions in the opposite orientation were unresponsive to the inducer. Tn3-Spice was also used to investigate the expression of a cluster of genes (hrp) which control pathogenicity and hypersensitivity elicited by Pseudomonas syringae pv. phaseolicola. An inducible region was identified which is expressed at low levels in vitro but becomes activated when the bacteria come into contact with the susceptible host, bean. Activation of this region occurred within 2 h post inoculation and was nearly complete by the time the bacteria began to multiply in the leaf tissue. The inaZ reporter appears to be at least 10(5)-fold more sensitive than lacZ in P.s.phaseolicola. Thus, the inaZ fusion system provides a sensitive, convenient and inexpensive tool for the study of bacterial gene expression, particularly during plant pathogenesis, and should be generally useful as a reporter gene system in Gram-negative bacteria. PMID- 2548842 TI - Differential signalling potential of insulin- and IGF-1-receptor cytoplasmic domains. AB - The human receptors for insulin-like growth factor 1 (IGF-1) and insulin, and two chimeric receptors consisting of ligand-binding, extracellular insulin receptor and intracellular IGF-1 receptor structures, have been expressed in NIH-3T3 fibroblasts. All four receptor types were synthesized, processed and transported to the cell surface to form high-affinity binding sites. All normal and chimeric receptors had an active tyrosine kinase which was regulated by homologous or heterologous ligands respectively. In addition, cell surface receptors were internalized efficiently and subjected to accelerated degradation in the presence of ligand. While all four types of receptor stimulated glucose transport with similar efficiency, they displayed significant differences in their mitogenic signalling potentials. Receptors with an IGF-1 receptor cytoplasmic domain were 10 times more active in stimulating DNA synthesis than the insulin receptor. In NIH-3T3 cells overexpressing wild-type and chimeric receptors, maximal growth responses obtained with IGF-1 or insulin alone were equivalent to those obtained with 10% fetal calf serum. We conclude that in the cell system employed the receptors for IGF-1 and insulin mediate short-term responses similarly, but display distinct characteristics in their long-term mitogenic signalling potentials. PMID- 2548843 TI - Bovine papilloma virus encoded E2 protein activates lymphokine genes through DNA elements, distinct from the consensus motif, in the long control region of its own genome. AB - Activation of T cells by antigen, lectin or a combination of phorbol ester (PMA) and calcium ionophore (A23187) leads to the induction of a set of lymphokine genes. Transfection of a human T cell leukemia cell line, Jurkat, or an African green monkey kidney cell line, CV1, with a cDNA encoding E2 protein, a trans activator of bovine papilloma virus type 1, results in activation of interleukin 2 (IL-2), interleukin 3 (IL-3) and granulocyte/macrophage colony-stimulating factor (GM-CSF) genes in a transient transfection assay. 5' deletion and mutation analyses showed that the sequence between positions -60 and a TATA-like sequence is required for basic promotor function and that the sequence between positions 95 and -73 containing conserved lymphokine element 2 (CLE2) and a GC box (CLE2/GC box) mediates the positive response to E2 protein. The latter has been previously shown to respond to PMA/A23187 stimulation or to p40tax, a trans-activator encoded by human T cell leukemia virus type 1 (HTLV-I). The sequence located between -108 and -99 (CLE1) is inhibitory to E2 protein or PMA/A23187 stimulation. The combination of E2 protein and PMA/A23187 appears to eliminate an inhibitory effect of the upstream region. However, E2 protein, like p40tax, mediates a positive response through CLE1 alone linked to the basic promoter sequence. The level of activation of the long control region (LCR) by E2 protein is unaffected by the number of CLE2/GC box sequences.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548845 TI - Transcription factors AP-3 and AP-2 interact with the SV40 enhancer in a mutually exclusive manner. AB - The 'core' sequence is critical for efficient transcriptional activity of the SV40 enhancer. Moreover, the core was shown to be involved in a signal transduction pathway elicited by treatment of cells with phorbol ester tumor promoters. We report here the identification and characterization of activator protein-3 (AP-3), which recognizes the core element. AP-3 was purified to near homogeneity and identified as a 48K polypeptide. The purified protein is an efficient transcriptional activator in vitro. In addition, we show that AP-3 and a second factor that recognizes the SV40 enhancer, AP-2, interact in a mutually exclusive manner. These studies should facilitate understanding of the mechanism by which the SV40 enhancer achieves its characteristic broad cell-type specificity. PMID- 2548844 TI - The Epstein-Barr virus early protein EB1 activates transcription from different responsive elements including AP-1 binding sites. AB - When expressed in Epstein-Barr virus (EBV) latently infected B cells, the EBV early protein EB1 trans-activates as many EBV early genes as does TPA. Several EB1 responsive elements (ZRE) have been identified in EBV early promoters and are located at relatively short distances from the TATA box. One of them (ZRE-M) overlaps with a consensus TPA responsive element (TRE) defined as an AP-1/c-jun/c fos binding site and is located in an EBV promoter controlling the expression of the post-transcriptional activator EB2. Another (ZREZ) is located in the promoter controlling the expression of EB1 and does not respond to TPA. These two ZREs have no apparent sequence homology. Although EB1 activates transcription from the AP-1 enhancer sequence and from the ZREZ, the activation is severely impaired by distance, suggesting that EB1 is more likely to be a promoter factor than an enhancer factor. These properties also suggest that EB1 is not functionally related to c-jun and c-fos. However, since EB1 can activate transcription from AP 1 binding sites when properly positioned, the role of this factor in the oncogenic properties of EBV should be considered. PMID- 2548846 TI - Tight folding of a passenger protein can interfere with the targeting function of a mitochondrial presequence. AB - The first seven residues of the yeast cytochrome oxidase subunit IV presequence are insufficient to target attached mouse dihydrofolate reductase into isolated yeast mitochondria. However, the targeting function of this truncated presequence can be restored by presenting the fusion protein to isolated mitochondria either as nascent, unfolded chains, or as full-length chains whose dihydrofolate reductase moiety had been destabilized either by urea treatment or by point mutations. The targeting efficiency of a mitochondrial presequence can thus be strongly influenced by the conformation of the attached 'passenger protein'. These results also underscore the difficulty of defining a 'minimal' mitochondrial targeting signal. PMID- 2548847 TI - Structural factors that control conformational transitions and serotype specificity in type 3 poliovirus. AB - The three-dimensional structure of the Sabin strain of type 3 poliovirus has been determined at 2.4 A resolution. Significant structural differences with the Mahoney strain of type 1 poliovirus are confined to loops and terminal extensions of the capsid proteins, occur in all of the major antigenic sites of the virion and typically involve insertions, deletions or the replacement of prolines. Several newly identified components of the structure participate in assembly dependent interactions which are relevant to the biologically important processes of viral assembly and uncoating. These include two sites of lipid substitution, two putative nucleotides and a beta sheet formed by the N-termini of capsid proteins VP4 and VP1. The structure provides an explanation for the temperature sensitive phenotype of the P3/Sabin strain. Amino acids that regulate temperature sensitivity in type 3 poliovirus are located in the interfaces between promoters, in the binding site for a lipid substituent and in an assembly-dependent extended beta sheet that stabilizes the association of pentamers. Several lines of evidence indicate that these structural components also control conformational transitions at various stages of the viral life cycle. PMID- 2548848 TI - Intermediates in Hin-mediated DNA inversion: a role for Fis and the recombinational enhancer in the strand exchange reaction. AB - The site-specific inversion reaction controlling flagellin synthesis in Salmonella involves the function of three proteins: Hin, Fis and HU. The DNA substrate must be supercoiled and contain a recombinational enhancer sequence in addition to the two recombination sites. Using mutant substrates or modified reaction conditions, large amounts of complexes can be generated which are recognized by double-stranded breaks within both recombination sites upon quenching. The cleaved molecules contain 2-bp staggered cuts within the central dinucleotide of the recombination site. Hin is covalently associated with the 5' end while the protruding 3' end contains a free hydoxyl. We demonstrate that complexes generated in the presence of an active enhancer are intermediates that have advanced past the major rate limiting step(s) of the reaction. In the absence of a functional enhancer, Hin is also able to assemble and catalyze site specific cleavages within the two recombination sites. However, these complexes are kinetically distinct from the complexes assembled with a functional enhancer and cannot generate inversion without an active enhancer. The results suggest that strand exchange leading to inversion is mediated by double-stranded cleavage of DNA at both recombination sites followed by the rotation of strands to position the DNA into the recombinant configuration. The role of the enhancer and DNA supercoiling in these reactions is discussed. PMID- 2548849 TI - Directionality of DnaA protein/DNA interaction. Active orientation of the DnaA protein/dnaA box complex in transcription termination. AB - The complex of DnaA protein with its 9 bp consensus binding site, the dnaA box 5' TT(A/T)T(A/C)CA(A/C)A, blocks transcribing RNA polymerase. In a model system, the rate of transcription was monitored distal to the dnaA box 5'-TTTTCCACA by the expression of a reporter gene. DnaA-dependent transcription termination occurred irrespective of whether the dnaA box region was or was not translated. Only the dnaA box orientation 5'-TTTTCCACA on the non-coding strand, but not the reverse orientation, was active in termination. This suggests that DnaA protein contacts only one strand of the DNA duplex. Oligonucleotide-directed mutation of a dnaA box present within the dnaA coding region resulted in increased expression of dnaA. This demonstrates that DnaA protein-directed transcription termination is an element of the autoregulation of the dnaA gene. PMID- 2548851 TI - Aspartic acids 96 and 85 play a central role in the function of bacteriorhodopsin as a proton pump. AB - A spectroscopic and functional analysis of two point-mutated bacteriorhodopsins (BRs) from phototrophic negative halobacterial strains is reported. Bacteriorhodopsin from strain 384 contains a glutamic acid instead of an aspartic acid at position 85 and BR from strain 326 contains asparagine instead of aspartic acid at position 96. Compared to wild-type BR, the M formation in BR Asp85---Glu is accwelerated approximately 10-fold, whereas the M decay in BR Asp96---Asn is slowed down approximately 50-fold at pH6. Purple membrane sheets containing the mutated BRs were oriented and immobilized in polyacrylamide gels or adsorbed to planar lipid films. The measured kinetics of the photocurrents under various conditions agree with the observed photocycle kinetics. The ineffectivity of BR Asp85---Glu resides in the dominance of an inactive species absorbing maximally at approximately 610 nm, while BR Asp96---Asn is ineffective due to its slow photocycle. These experimental results suggest that aspartic acid 96 plays a crucial role for the reprotonation of the Schiff base. Both residues are essential for an effective proton pump. PMID- 2548850 TI - Molecular cloning of a maize gene involved in photosynthetic membrane organization that is regulated by Robertson's Mutator. AB - The maize photosynthetic mutant hcf106 has a distinctive and unusual thylakoid membrane organization, and fails to accumulate three of the five thylakoid membrane protein complexes. This mutant arose in a Robertson's Mutator background, and shows somatic instability typical of a transposon-induced allele. In addition, hcf106 is suppressed when Mu1 elements are inactive and modified in their terminal inverted repeats. Thus plants homozygous for the mutant allele adopt a mutant phenotype only when Mu1 elements are active and unmodified. DNA from the mutant allele has been cloned by 'transposon-tagging' using the transposon Mu1, and the identity of the clone confirmed by observing somatic excision of the transposon in a revertant sector. A 1.2 kb transcript homologous to the cloned DNA is found in wild-type and suppressed seedlings, but is not found in mutant seedlings, suggesting that suppression is mediated at the level of transcript accumulation. PMID- 2548852 TI - GABAA receptor beta subunit heterogeneity: functional expression of cloned cDNAs. AB - Cloned cDNAs encoding two new beta subunits of the rat and bovine GABAA receptor have been isolated using a degenerate oligonucleotide probe based on a highly conserved peptide sequence in the second transmembrane domain of GABAA receptor subunits. The beta 2 and beta 3 subunits share approximately 72% sequence identity with the previously characterized beta 1 polypeptide. Northern analysis showed that both beta 2 and beta 3 mRNAs are more abundant in the brain than beta 1 mRNA. All three beta subunit encoding cDNAs were also identified in a library constructed from adrenal medulla RNA. Each beta subunit, when co-expressed in Xenopus oocytes with an alpha subunit, forms functional GABAA receptors. These results, together with the known alpha subunit heterogeneity, suggest that a variety of related but functionally distinct GABAA receptor subtypes are generated by different subunit combinations. PMID- 2548853 TI - Inhibition of insulin-dependent lipogenesis and anti-lipolysis by protein tyrosine kinase inhibitors. AB - Protein tyrosine kinase (PTK) blockers which competitively inhibit the kinase activity of insulin receptors were synthesized and their properties examined. The best insulin receptor kinase (IRK) inhibitors possess either one hydroxyphenyl ring and two carboxyl groups or two phenyl rings and one carboxyl group. All the inhibitors, except tBoc-tyrosine aminomalonate, effectively block the IRK catalyzed phosphorylation of exogenous substrate, but only partially block receptor autophosphorylation. These PTK blockers inhibit the insulin induced [14C]glucose assimilation into lipids (lipogenesis), but fail to inhibit the anti lipolytic effect of the hormone. Only tBocTyr-aminomalonate was found to inhibit all the effects of insulin measured: insulin-stimulated phosphorylation of exogenous substrate, IRK autophosphorylation, insulin-dependent lipogenesis and the insulin-dependent anti-lipolytic effect. This inhibitor is the first blocker which is reported to block insulin-dependent anti-lipolysis. The inhibitors examined are devoid of general adverse effects since they have no effect on insulin-independent lipolysis, on [U14C]fructose assimilation or on ( )isoproterenol-stimulated lipolysis. These studies suggest that insulin-dependent lipogenesis and anti-lipolysis may be mediated by two distinguishable signalling pathways. This study also suggests that PTK inhibitors may become useful tools in the investigation of the signalling pathways of PTKs. PMID- 2548854 TI - Topology of the non-structural rotavirus receptor glycoprotein NS28 in the rough endoplasmic reticulum. AB - The rotavirus non-structural glycoprotein (NS28), the receptor for the virus core during budding into the lumen of the rough endoplasmic reticulum (RER), is 175 amino acids long and possesses an uncleaved signal sequence and two amino terminal glycosylation sites. Utilizing one of three potential hydrophobic domains, the protein spans the membrane only once, with the glycosylated amino terminal region oriented to the luminal side of the ER and the carboxy-terminal region to the cytoplasmic side. To localize sequences involved in translocation of NS28, we constructed a series of mutations in the coding regions for the hydrophobic domains of the protein. Mutant protein products were studied by in vitro translation and by transfection in vivo. In transfected cells, all mutant forms localize to the ER, and none are secreted. In vitro, each of the three hydrophobic domains is able to associate with microsomes. However, glycosylation and proteolysis of wild-type and mutant forms of NS28 indicates that the wild type protein is anchored in the membrane only by the second hydrophobic domain, leaving approximately 131 residues exposed on the cytoplasmic side for receptor - ligand interaction. PMID- 2548855 TI - The human L-myc gene is expressed as two forms of protein in small cell lung carcinoma cell lines: detection by monoclonal antibodies specific to two myc homology box sequences. AB - The L-myc gene is the third member of the myc family of proto-oncogenes. Amplification and elevated expression of the L-myc gene has been detected in a subset of small cell lung carcinoma (SCLC) cell lines. The biological properties and functions of the L-myc gene and its product have not yet been elucidated. Monoclonal antibodies against two myc homology boxes were used to characterize the L-myc gene product. These antibodies react with two groups of polypeptides of apparent masses of 60, 61 and 66 kd (the long forms), and 34 and 37 kd (the short forms) in SCLC cells expressing L-myc transcripts. The long form L-myc proteins are associated with the nuclear fraction of the cells. The short form L-myc proteins are present in the cytoplasmic fraction, though diffusion of the short forms from the nucleus during cell fractionation cannot be ruled out. The half life of the long form polypeptides is approximately 45-90 min. The short form polypeptides have a half-life of approximately 120-180 min. The L-myc protein is not detectable in the mitotic cells, suggesting that the L-myc protein expression is tightly regulated during the cell cycle. PMID- 2548856 TI - A point mutation in the CYC1 UAS1 creates a new combination of regulatory elements that activate transcription synergistically. AB - Dissection of the upstream activation site 1 (UAS1) of the yeast CYC1 gene showed that the A and B regions respond individually to regulation by the HAP1 protein, and that a point mutation in the B region converts this region to a translation upstream factor (TUF)-regulated element. Combinatorial analyses revealed that the transacting factors involved with these wild-type and mutant UAS1 target sites combine to activate transcription in a synergistic manner. Furthermore, combinations of heterologous factors, made possible by the point mutation, create a new specificity of regulation that differs from regulation by any one factor individually. PMID- 2548857 TI - Epstein-Barr virus bicistronic mRNAs generated by facultative splicing code for two transcriptional trans-activators. AB - The Epstein-Barr virus (EBV) genome codes for several transcriptional trans activators. One of them, the BZLF1 open reading frame (ORF)-encoded product EB1, is able to induce the productive cycle in infected B cells. From the cloning and characterization of full-length cDNAs, we found that EB1 could be made from three overlapping messenger RNAs expressed under the control of two different promoters that we call P1 and P2. The first mRNA, 1 kb long, is made from the P1 promoter and codes for EB1 alone. The two other mRNAs, respectively 3 and 4 kb long and made by facultative splicing, are bicistronic mRNAs. They code not only for the trans-activator EB1 but also for a second EBV transcriptional trans-activator R, encoded by the BRLF1 ORF. In effect, authentic EB1 and R proteins are expressed from the 3 and 4 kb long cDNAs as demonstrated by identification of the proteins with specific antisera. In addition, EB1 and R expressed from the 3 and 4 kb cDNAs activate transcription from their specific targets in the EBV early promoter DR. PMID- 2548858 TI - DNA topoisomerase II activity in nonreplicating, transcriptionally inactive, chicken late spermatids. AB - To study a possible differential involvement of type I and type II DNA topoisomerases in the functional and structural changes that chromatin undergoes during spermatogenesis, we have determined both enzymatic activities in chicken testis cell nuclei at successive stages of differentiation. Whereas DNA topoisomerase I varies in parallel with transcriptional activity, DNA topoisomerase II was present in both replicating, transcriptionally active chicken testis cells and nonreplicating, transcriptionally inactive late spermatids. The presence of DNA topoisomerase II activity in late spermatids and, in addition, the relative increment of drug-induced topo-II-mediated DNA cleavage detected in these cells, suggest that DNA topoisomerase II might modulate the topology of DNA during the marked changes that chromatin structure undergoes in the nucleohistone-nucleoprotamine transition at the end of the spermiogenesis and could be involved in the final organization of DNA within the nucleus of the male gamete. PMID- 2548859 TI - DNA gyrase can supercoil DNA circles as small as 174 base pairs. AB - DNA gyrase introduces negative supercoils into closed-circular DNA using the free energy of ATP hydrolysis. Consideration of steric and thermodynamic aspects of the supercoiling reaction indicates that there should be a lower limit to the size of DNA circle which can be supercoiled by gyrase. We have investigated the supercoiling reaction of circles from 116-427 base pairs (bp) in size and have determined that gyrase can supercoil certain relaxed isomers of circles as small as 174 bp, dependent on the final superhelix density of the supercoiled product. Furthermore, this limiting superhelical density (-0.11) is the same as that determined for the supercoiling of plasmid pBR322. We also find that although circles in the range 116-152 bp cannot be supercoiled, they can nevertheless be relaxed by gyrase when positively supercoiled. These data suggest that the conformational changes associated with the supercoiling reaction can be carried out by gyrase in a circle as small as 116 bp. We discuss these results with respect to the thermodynamics of DNA supercoiling and steric aspects of the gyrase mechanism. PMID- 2548860 TI - Complete foldback transposable elements encode a novel protein found in Drosophila melanogaster. AB - An apparently complete foldback (FB) transposable element homologous to FB white crimson (FBwc) was analyzed. A complete FB element could encode one or more proteins required for regulation of FB transposition. The central DNA region (the loop) and the junctions between the loop and the inverted terminal repeats were sequenced. Three open reading frames (ORFs) are present in the loop, and a novel 308 bp inverted repeat is present at the junctions. No significant homologies were found when the DNA sequences of the loop region and the novel inverted repeat were screened against the Gene data bank. Antibodies were prepared in guinea-pigs against a peptide present near the amino terminus of ORF1, the longest ORF. A 71,000 dalton protein was isolated from an extract of Drosophila melanogaster early-stage embryos on an anti-ORF1 peptide-affinity column. Immunohistochemical studies of adult flies demonstrate localization of this protein in egg chambers. PMID- 2548861 TI - Morphological and neoplastic transformation of C3H/10T1/2 Cl 8 mouse embryo cells by insoluble carcinogenic nickel compounds. AB - We studied induction of cytotoxicity and morphological transformation in C3H/10T1/2 Cl 8 (10T1/2) mouse embryo fibroblasts by soluble and insoluble carcinogenic nickel compounds. Soluble nickel sulfate and nickel chloride caused dose-dependent cytotoxicity in the concentration range from 0.5 microM to 100 microM after 48 hr treatments, but neither compound induced morphological transformation even at concentrations causing up to 94% cytotoxicity. Insoluble nickel subsulfide, nickel monosulfide, and nickel oxide caused dose-dependent cytotoxicity and a low, dose-dependent frequency of morphological transformation in the concentration ranges from 0.5 to 40 microM, 5 to 50 microM, and 50 to 400 microM, respectively, after 48 hr exposure of cells to these compounds. Foci were predominantly of type II morphology; type III foci were rare. The insoluble nickel compounds studied caused no induction of base substitution mutations to ouabain resistance in 10T1/2 cells over concentration ranges that induced morphological transformation. Nickel subsulfide and nickel monosulfide were taken into cells by phagocytosis, since particles were visible in intracytoplasmic vacuoles. Numerous nickel oxide particles were found associated with cells, but true phagocytic uptake was difficult to detect since no vacuoles were observed. We twice cloned type II and type III foci induced by insoluble nickel compounds, established independent cell lines, and characterized their phenotypes. Four of seven of these cell lines had three- to fourfold increased saturation densities compared to 10T1/2 cells, formed type II and type III foci in reconstruction assays, and grew in soft agarose. One cell line induced by nickel oxide formed tumors in nude mice. These data indicate that insoluble carcinogenic nickel compounds induced type II foci in 10T1/2 cells, some of which were tumorigenic, and that the 10T1/2 cell system is suitable for studying mechanisms of nickel compound-induced morphological transformation in mammalian cells. PMID- 2548862 TI - The effect of induced alkalosis and acidosis on plasma lactate and work output in elite oarsmen. AB - In order to test the effect of artificially induced alkalosis and acidosis on the appearance of plasma lactate and work production, six well-trained oarsmen (age = 23.8 +/- 2.5 years; mass = 82.0 +/- 7.5 kg) were tested on three separate occasions after ingestion of 0.3 g.kg-1. NH4Cl (acidotic), NaHCO3 (alkalotic) or a placebo (control). Blood was taken from a forearm vein immediately prior to exercise for determination of pH and bicarbonate. One hour following the ingestion period, subjects rowed on a stationary ergometer at a pre-determined sub-maximal rate for 4 min, then underwent an immediate transition to a maximal effort for 2 min. Blood samples from an indwelling catheter placed in the cephalic vein were taken at rest and every 30 s during the 6 min exercise period as well as at 1, 3, 6, 9, 12, 15, 18, 21, 25 and 30 min during the passive recovery period. Pre-exercise blood values demonstrated significant differences (p less than 0.01) in pH and bicarbonate in all three conditions. Work outputs were unchanged in the submaximal test and in the maximal test (p greater than 0.05), although a trend toward decreased production was evident in the acidotic condition. Analysis of exercise blood samples using ANOVA with repeated measures revealed that the linear increase in plasma lactate concentration during control was significantly greater than acidosis (p less than 0.01). Although plasma lactate values during alkalosis were consistently elevated above control there was no significant difference in the linear trend (p greater than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548863 TI - Muscle bio-energetics in acute glycolytic block: in vivo phosphorus-nuclear magnetic resonance study of iodo-acetate injected rats. AB - In vivo phosphorus nuclear magnetic resonance spectroscopy of muscle was performed at rest, during work and during postexercise recovery in rats injected with iodo-acetate (IA) (35-40 mg.kg-1, intra-arterially), in order to follow bio energetic changes in muscle with acute glycolytic block. Three animals with contracture had very low ratios of phosphocreatine:inorganic phosphate (PCr:Pi) at rest (0.5-0.9). The PCr:Pi were normal at rest (6.9 +/- 2.0, +/- 2 SD) in all other rats. Exercise-induced continuous accumulation of phosphomonoesters (PME), the characteristic finding of glycolytic block, was observed. The end-exercise levels of PME correlated with the degree of block measured in vitro. During steady-state work, induced by nerve stimulation at four frequencies, PCr:Pi values were significantly lower (p less than 0.02) than the control values at 0.25, 1.0 and 2.0 Hz. The ATP levels fell during exercise to reach 75% +/- 7% of initial values. The recovery of PCr:Pi from exercise and the disappearance of PME were slow. Two animals which survived the IA injection demonstrated much lower PME accumulation 18 h later. It is concluded that in acute muscle glycolytic block: (1) energy metabolism is impaired during exercise and also at rest, (2) accumulating PME can serve as an indicator of the degree of glycolytic block, (3) ATP levels fall during work, and (4) postexercise recovery is slow. The findings are compared with 31P-NMR observations in chronic muscle glycolytic disorders. PMID- 2548864 TI - Effects of treadmill running on plasma beta-endorphin, corticotropin, and cortisol levels in male and female 10K runners. AB - Reports of plasma beta-endorphin (B-EN) levels in response to submaximal exercise have been highly disparate. Variations in experimental design have complicated interpretation of previous research. The present study was designed to determine whether a sequential change in plasma beta-endorphin (B-EN), corticotropin (ACTH), and cortisol levels occurs in response to a 30-min submaximal run. Twenty three subjects were divided into four groups: male runners, female runners, sedentary males and sedentary females. Subjects ran on a treadmill at 80% of previously determined maximum heart rate. Five plasma samples were obtained through an indwelling catheter before exercise (-30 and 0 min), at 15 and 30 min of exercise, and after 30 minutes of recovery. The run resulted in no rise in B EN, ACTH, and cortisol despite an elevated rectal temperature. B-EN values were significantly higher in males than in females (p less than 0.01). No sex or training differences were seen with respect to change of hormone concentrations over the course of the run. Three male runners developed symptoms of vasovagal syncope after the catheter placement and had high initial B-EN, ACTH, and cortisol concentrations which decreased throughout the run. These data indicate that gender and training do not affect ACTH and cortisol concentrations before, during, and after 30 min of treadmill running at 80% of maximum heart rate, whereas B-EN concentrations are higher in males under these conditions. PMID- 2548865 TI - Quality control parameters and interpretive criteria for in vitro susceptibility tests with the macrolide azithromycin. Collaborative Antimicrobial Susceptibility Testing Group. AB - Quality control parameters for broth microdilution and disk diffusion susceptibility tests were defined and the interpretive criteria for disk diffusion tests reviewed. For interpretation of tests with 15 micrograms azithromycin disks, the following criteria are recommended: greater than or equal to 19 mm for the susceptible category (MIC less than or equal to 2.0 micrograms/ml) and less than or equal to 15 mm for the resistant category (MIC greater than or equal to 8.0 micrograms/ml). Using these criteria, there was 97% overall agreement between broth dilution and disk diffusion tests; Haemophilus influenzae isolates were susceptible to azithromycin by both methods. The quality control strain Staphylococcus aureus ATCC 25923 gave zones of 21 to 26 mm in diameter in a six-laboratory collaborative study. In azithromycin broth microdilution tests the following MIC control limits are recommended: Escherichia coli ATCC 25922, 2.0-8.0 micrograms/ml; Staphylococcus aureus ATCC 29213, 0.25 1.0 micrograms/ml; and Enterococcus faecalis ATCC 29212, 1.0-4.0 micrograms/ml. PMID- 2548866 TI - Comparative antibacterial activity of the new cephalosporin cefcanel against anaerobic bacteria. AB - The activity of cefcanel against anaerobic cocci, Clostridium perfringens, Bacteroides fragilis, Bacteroides spp. and fusobacteria was determined by the agar dilution method and compared with the activity of cefaclor, cephalexin, cefadroxil, phenoxymethylpenicillin and ampicillin. Cefcanel showed good activity against Clostridium perfringens, Bacteroides spp. and fusobacteria (MIC90 = 1-4 mg/l). Against anaerobic cocci its MIC90 value was 16 mg/l, and against Bacteroides fragilis, 32 mg/l. Cefcanel has an antibacterial activity that warrants investigation in clinical trials. PMID- 2548867 TI - Treatment of chronic experimental Staphylococcus aureus osteomyelitis with LY146032 and vancomycin. AB - LY146032 and vancomycin were compared as therapeutic agents in the treatment of chronic Staphylococcus aureus osteomyelitis in the rat. Quantitative cultures disclosed that one of 16, none of 16 and two of 17 tibiae were sterile from the control LY146032, and vancomycin groups, respectively. From positive cultures, geometric mean staphylococcal CFU per gram of bone were as follows: control, 5.13 +/- 1.58; LY146032, 5.36 +/- 0.43 (p = 0.57); and vancomycin, 4.33 +/- 1.73 (p = 0.078). Mean gross pathology was decreased significantly in both treatment groups. LY146032 was no more effective than vancomycin in reducing bacterial counts. PMID- 2548869 TI - Low dose 1-hexylcarbamoyl-5-fluorouracil (HCFU) recommended for cirrhotic patients with hepatocellular carcinoma. AB - The metabolism of 1-hexylcarbamoyl-5-fluorouracil (HCFU), a drug prescribed for treating patients with hepatocellular carcinoma (HCC), was studied in relation to liver function, with the objective of clarifying the occurrence of any adverse side-effects on the central nervous system. Twenty-five HCC patients were administered 3.4 mg/kg HCFU once orally, after which the blood levels of HCFU and its derivatives (5-FU, CPEFU, CPRFU, HHCFU, OHCFU and F-beta-alanine) were serially measured using high performance liquid chromatography. The area under the concentration curve (AUC) of HCFU in the group of ICG R15 greater than or equal to 30% (group 2) was 5.35 +/- 1.73 h.micrograms/ml, a value which was significantly higher than the 2.60 +/- 1.19 h.micrograms/ml recorded for the group of ICG R15 less than 30% (group 1) (P less than 0.001). The AUC of HCFU had a significant positive correlation with the value of ICG R15 (P = 0.002) or the serum total bilirubin (P = 0.0005). The AUC of 5-FU showed no difference between the two groups. The AUC of CPRFU in group 2 was 0.16 +/- 0.25 h.micrograms/ml, a value significantly lower than the 0.48 +/- 0.39 h.micrograms/ml in group 1 (P = 0.023). There was no correlation between the AUC of other derivatives and the markers of liver function. These data suggest that, in patients with advanced cirrhosis, the accumulation of HCFU is related to the occurrence of side-effects from the administered drug, ingested over a long-term period. Therefore, when HCFU is given to cirrhotic patients with both HCC and 30% or more ICG R15, a careful monitoring for side-effects is required. PMID- 2548868 TI - New perspectives on the molecular pharmacology of affective disorders. AB - Research with antidepressants has emphasized the importance of a delayed deamplification of the linked serotonin (5HT)/norepinephrine (NE) receptor coupled adenylate cyclase system in brain. The basic phenomena of regulation of receptor number and function of the beta adrenoceptor linked adenylate cyclase system in brain are well established, with NE regulating beta adrenoceptors in the high agonist affinity conformation (linked to adenylate cyclase and down regulated by antidepressants), and with 5HT regulating those receptors in the low agonist affinity conformation. The biochemical effector systems of NE and 5HT are discussed and it is concluded that the final common pathway of signal transduction is protein kinase mediated phosphorylation of cellular proteins. Glucocorticoid receptors are located in the perikarya of aminergic cell bodies and may exert their effects by modifying the genomic expression of the diffusely projecting stress-responsive monoamine systems. The molecular neurobiology of beta adrenoceptors, with its implication for genetic and immunologic investigations, is briefly discussed and further research on stimulus transcription coupling and regulation of gene expression in brain is suggested as an exciting new direction in central receptor research relevant to the psychopharmacology of affective and other disorders. PMID- 2548870 TI - In vitro and in vivo evaluation of the indoloquinone EO-9 (NSC 382 459) against human small cell carcinoma of the lung. AB - As the indoloquinone EO-9 has previously shown activity in several tumor model systems it was evaluated against four human small cell lung cancer cell lines by the clonogenic assay. In two cell lines (Nyh and Tol), exponential dose-response curves were achieved with both 1 h and continuous exposure, whereas no cell kill was obtained in the other two cell lines (69 and 592) when tested with 1 h incubation up to 0.25 microgram/ml. When the cells were exposed to drug in vitro, flow cytometric DNA analysis showed perturbations in the cell cycle distribution of the most sensitive cell line (Tol) at a lower EO-9 concentration than in the less sensitive cel line (592). This in vitro predicted difference in EO-9 sensitivity between two of the cell lines (592 and Tol) was confirmed when the cell lines were heterotransplanted to nude mice. PMID- 2548871 TI - Elevation of a novel pituitary protein (7B2) in the plasma in small cell carcinoma of the lung. AB - 7B2 is a new protein isolated from human pituitary glands and distributed widely, particularly with high concentrations in the neuroendocrine tissues in the rat. We measured plasma 7B2 concentrations in 333 normal subjects, 20 patients with benign pulmonary disease and 111 patients with primary lung cancer (21 small cell carcinoma, 90 non-small cell carcinoma). A normal range for plasma 7B2 concentrations was defined as less than the mean + 3 S.D. value (110 pg/ml) based on plasma 7B2 concentrations in 333 normal subjects. Elevation of the plasma 7B2 concentration over the normal range was observed in 15 of 21 patients (71.4%) with small cell carcinoma, eight of 90 (8.9%) with non-small cell carcinoma and four of 20 (20%) with benign pulmonary disease. Plasma 7B2 concentrations correlated with the clinical course on chemotherapy in some patients with small cell carcinoma. Immunocytochemical studies revealed numerous 7B2-positive cells in the small cell carcinoma specimen, while 7B2 staining was not observed in the non-small cell carcinoma and the normal lung specimens. These findings suggest that 7B2 is secreted by the small cell carcinoma of the lung, which caused elevation of plasma 7B2 in these patients. 7B2 might be a possible plasma tumor marker for the small cell carcinoma of the lung. PMID- 2548872 TI - Prognostic value of estrogen and progesterone receptors in primary infiltrating ductal breast cancer. A sequential multivariate analysis of 1262 patients. AB - Nine prognostic variables were evaluated for their significance in predicting the overall survival (OS), the length of disease-free survival (DFS) and the length of metastasis-free survival (MFS) of 1262 patients with primary breast cancer. The variables studied were: UICC clinical stage; menopausal status; histologic grade; number of involved nodes; anatomic tumor size; estrogen and progesterone receptors; local and adjuvant therapies. Three sequential multivariate analyses, at 2, 5 and 10 years, using the Cox proportional hazard regression model, were carried out to identify those variables most highly related to the criteria studied (overall, disease-free, metastasis-free survivals) and especially to fully evaluate the effects of hormonal receptors on prognosis and their stability over time. Our results showed that number of involved nodes and histologic grade were the most significant prognostic factors for all periods of time and whatever the criterion studied; ER had no predictive value while PR was an independent prognostic factor for metastasis-free survival at 2 years (P = 0.01) and 5 years (P = 0.02) but lost its significance at 10 years (P = 0.06). In the subgroup of 261 patients who received prolonged post-operative adjuvant chemotherapies, PR was the main prognostic factor for MFS at 2 years (P = 0.03) and the second at 5 years (P = 0.05) just after number of involved nodes. In the 1001 patients who did not receive prolonged post-operative adjuvant chemotherapies ER was significant for MFS at 5 and 10 years. The present data urge the need for a periodic redefinition of prognostic factors in primary breast cancer. PMID- 2548873 TI - Release of PAF-acether and eicosanoids from guinea-pig alveolar macrophages by FMLP: effect of cyclo-oxygenase and lipoxygenase inhibition. AB - Adherent guinea-pig alveolar macrophages stimulated in vitro with FMLP (1 microM) for 5-60 min released PAF-acether, TXB2 and LTB4, with maximal levels being achieved after a 5 min incubation. The levels of eicosanoids were maintained during the 60 min incubation period, whereas the levels of PAF-acether decreased and were no longer detectable after 60 min incubation. Indomethacin (1 microM) completely suppressed the release of TXB2 stimulated by FMLP (1 microM for 5 min) whilst potentiating the release of LTB4. The selective 5-lipoxygenase inhibitor, BW A137C (1 microM), abolished the stimulated release of LTB4 without affecting the release of TXB2. BW755C (1-20 microM) caused a dose-related inhibition of TXB2 release but did not affect LTB4 release, while none of the drugs studied affected the release of PAF-acether. These results indicate that the FMLP-induced release of PAF-acether from guinea-pig alveolar macrophages is not susceptible to modulation by selective inhibition of lipoxygenase or cyclo-oxygenase and is therefore not secondary to the synthesis of LTB4 or TXB2. PMID- 2548874 TI - Mass changes of inositol(1,4,5)trisphosphate in trachealis muscle following agonist stimulation. AB - Agonist-stimulated changes in D-Ins(1,4,5)P3 has been examined in tracheal smooth muscle using a specific radioreceptor assay utilizing the binding of [3H] or [32P]Ins(1,4,5)P3 to a bovine adrenal cortex preparation. Carbachol produced a maximal increase in Ins(1,4,5)P3 concentration after 5 s (basal: 12.9 +/- 0.8; stimulated: 27 +/- 1.5 pmol/mg protein) with an EC50 of 6.7 +/- 0.8 microM. With longer stimulation periods Ins(1,4,5)P3 rapidly returned to basal values by 30 s. This is the first report of mass measurements of Ins(1,4,5)P3 in smooth muscle. PMID- 2548875 TI - Ethanol suppresses hippocampal cell firing through a calcium and cyclic AMP sensitive mechanism. AB - The effects of ethanol were studied intracellularly in hippocampal pyramidal cells in vitro. Ethanol, 50-100 mM, produced a marked suppression of neuronal firing. This effect was blocked by treating the cell with cyclic 3', 5'-adenosine monophosphate (cAMP) or cadmium ions. Ethanol had no effect on the after hyperpolarizing current. It is concluded that the ethanol-induced reduction of firing rate is due to a calcium-dependent process, and modulated by cAMP. PMID- 2548876 TI - Vasoconstrictor-induced heterologous down-regulation of vascular atrial natriuretic peptide receptor. AB - Long-term (24 h) pretreatment of cultured rat vascular smooth muscle cells with 100 nM angiotensin and 1 microM vasopressin induced a marked reduction of the maximal binding capacity of atrial natriuretic peptide (ANP) receptors in a fashion similar to that induced by phorbol ester. The down-regulation of the receptors induced by vasoconstrictors and phorbol ester was concomitantly associated with an attenuation of ANP-stimulated cGMP accumulation. These data suggest that vasoconstrictor-induced activation of protein kinase C is involved in the mechanism of heterologous down-regulation of vascular ANP receptors. PMID- 2548878 TI - The effects of non-competitive NMDA receptor antagonists on rats exposed to hyperbaric pressure. AB - The high pressure neurological syndrome (HPNS) occurs when man or animals are exposed to hyperbaric pressure. Four non-competitive N-methyl-D-aspartate (NMDA) antagonists - MK-801, phencyclidine (PCP), SKF 10,047 and ketamine were tested in rats for effects on the HPNS. All drugs were injected i.p. prior to compression; ketamine was also infused i.v. Control rats received saline. Rats were exposed individually to increasing helium pressure (PO2 0.5 atmospheres absolute ATA). Three endpoints were used to assess HPNS: onset pressures for tremor, myoclonus and convulsions. Neither MK-801 (0.03 and 0.3 mg/kg) nor SKF 10,047 (50 mg/kg) had any effect on the onset pressures for tremor, myoclonus or convulsions, although the type of seizure was modified from the clonic/tonic seizure seen in controls to purely clonic. PCP (5 mg/kg) had no effect on the endpoints, but pressure enhanced the excitation and stereotypy seen at 1 ATA. Ketamine (100 mg/kg i.p.) did not affect tremor or myoclonus; ketamine infused i.v. at pressure only prevented tremor and myoclonus at 'anaesthetizing' concentrations. Our results show that these non-competitive NMDA antagonists had little effect on HPNS, in contrast to competitive NMDA antagonists, such as AP7, which are highly effective. Possible explanations for this lack of effect include (1) interactions with NMDA receptor channels are pressure dependent; (2) other actions of these antagonists override their effects on the NMDA receptor channel. PMID- 2548877 TI - Modulation of mu-mediated antinociception by delta agonists in the mouse: selective potentiation of morphine and normorphine by [D-Pen2,D-Pen5]enkephalin. AB - The effect of the delta-selective agonist [D-Pen2,D-Pen5]enkephalin (DPDPE) on the antinociception produced by intracerebroventricular (i.c.v.) administration of the mu agonists morphine, [D-Ala2,NMePhe4,Gly-ol5]enkephalin (DAGO), [NMePhe3,D-Pro4]morphiceptin (PLO17), beta-endorphin, phenazocine, etorphine and sufentanil was studied in mice. Only the antinociceptive effects of morphine and normorphine were modulated by i.c.v. coadministration of a dose of DPDPE which did not produce any significant antinociception alone. Both the morphine and normorphine dose-response lines were displaced to the left in the presence of DPDPE. The delta-selective antagonist ICI174,864 (N,N-diallyl-Tyr-Aib-Aib-Phe-Leu OH) (where Aib is alpha-aminoisobutyric acid) blocked the modulation of morphine antinociception by DPDPE. ICI 174,864 alone failed to produce either a significant increase or decrease of morphine, phenazocine, etorphine or beta endorphin antinociception. The results of the present study provide support for the hypothesis that the enkephalins may function to modulate antinociception produced at the mu receptor; such modulation may come about via the existence of an opioid mu-delta receptor complex. The mu receptors existing in such a complex may be selectively activated by morphine and normorphine, but not the other mu agonists studied here. Thus, the enkephalins may function both to directly initiate, as well as to modulate, some forms of supraspinal mu receptor-mediated antinociception. PMID- 2548879 TI - Metaphit, an isothiocyanate analog of PCP, induces audiogenic seizures in mice. AB - Metaphit induces audiogenic seizures in mice. The most severe clonic/tonic seizures occur 18-24 h after metaphit administration. After 48 h the incidence of the seizure episodes begin to diminish. These audiogenic seizures can be prevented by the administration of either PCP or MK-801 24 h after metaphit and 30 min prior to audio stimulation. These seizures may be due to a modulation of the PCP recognition site by metaphit which results in an enhanced probability that the NMDA/PCP ion channels are open. PMID- 2548881 TI - (+)-Niguldipine binds with very high affinity to Ca2+ channels and to a subtype of alpha 1-adrenoceptors. AB - The enantiomers of the 1,4-dihydropyridine (DHP) niguldipine (3-methyl-5-[3-(4,4 diphenyl-1-piperidinyl)-propyl]- 1,4-dihydro-2,6-dimethyl-4-(3-nitrophenyl) pyridine-3,5-dicarboxylate- hydrochloride) were investigated with respect to their interaction with 1,4-DHP receptors on L-type Ca2+ channels and alpha adrenoceptors. The Ki values for niguldipine were dependent on the membrane protein concentrations in the radioligand binding assay. 'True' Ki values (at extrapolated 'zero' membrane protein) were determined with guinea-pig membranes for (+)-niguldipine and were found to be 85 pmol/l for the 1,4-DHP receptor of skeletal muscle, 140 pmol/l for that of brain and 45 pmol/l for that of heart. ( )-Niguldipine was approximately 40 times less potent. (+)-Niguldipine (Ki: 78 nmol/l) and (-)-niguldipine (Ki: 58 nmol/l) bound with approximately equal affinity to the alpha 1-adrenoceptors ('alpha 1B') in liver cell membranes. The (+)-niguldipine alpha 1-adrenoceptor inhibition data for rat brain cortex membranes were better fitted by a two-site model. The high-affinity component ('alpha 1A') had a Ki value of 52 pmol/l in competition experiments with [3H]prazosin. The low-affinity site (alpha 1B) had 200- to 600-fold less affinity. (-)-Niguldipine was greater than 40-fold less potent at alpha 1A- but was nearly equipotent to the (+)enantiomer at alpha 1B-sites. (+)-Niguldipine was the most selective compound for discriminating alpha 1A- from alpha 1B adrenoceptors and is a novel prototype for 1,4-DHPs which bind with nearly equal affinity to skeletal muscle and brain or heart 1,4-DHP receptors. PMID- 2548880 TI - GABAA receptors in the rat stomach may mediate mucoprotective effects. AB - The occurrence and characteristics of binding sites specific for gamma aminobutyric acid (GABA) and muscimol in the rat stomach were examined by biochemical and autoradiographic techniques, and the effects of GABAergic model compounds on gastric ulceration induced by chemical irritation was studied in intact and unilaterally vagotomized rats. Specific binding sites for [3H]GABA and [3H]muscimol, which showed the characteristics of GABAA receptors, were demonstrated on gastric membranes. Specific muscimol binding sites were found in all regions of the stomach and were present in both the mucosal layer and the remaining tissue of the stomach. Oral pretreatment of the rats with GABA, selective GABAA receptor agonists, or inhibitors of GABA degradation protected the gastric mucosa against the ulcers induced by acidified ethanol (chemical irritant), in both intact and vagotomized rats. These findings are consistent with the view that a subpopulation of GABAA receptors in the rat stomach may mediate the anti-ulcer effect. PMID- 2548883 TI - Memantine inhibits serotonin-induced rise of cytosolic Ca2+ activity and of cyclic GMP level in a neuronal cell line. AB - Serotonin (5-HT) evoked a rise of cytosolic Ca2+ activity in neuroblastoma X glioma hybrid cells, most probably due to the entry of extracellular Ca2+; cyclic GMP synthesis was also stimulated. The rise of both cytosolic Ca2+ activity and of cyclic GMP level was blocked by memantine (1-amino-3,5-dimethyladamantane). Memantine inhibited the rise of the cyclic GMP level non-competitively (Ki about 50 microM). Thus, memantine suppresses the effects of 5-HT in the neuronal cell line, most likely by blocking Ca2+-permeable ion channels. This interpretation is in line with the previously reported finding that memantine suppressed the 5-HT induced depolarizing response in the same cell line. PMID- 2548882 TI - Serotonin raises the cyclic GMP level in a neuronal cell line via 5-HT3 receptors. AB - Serotonin (5-HT) induced a transient rise of the cyclic GMP level in neuroblastoma X glioma hybrid cells, half-maximally at 1 microM 5-HT. 2-Methyl-5 HT displayed an about 5 times lower potency but equal efficacy. alpha-Methyl-5-HT and 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) were completely ineffective at concentrations up to 30 microM. Antagonists specific for 5-HT3 receptors, ICS 205-930, GR 38032 F and MDL 72222, blocked the response to 5-HT at nanomolar concentrations but antagonists directed towards 5-HT1 and 5-HT2 receptors, ketanserin and methysergide, had no effect at concentrations up to 1 microM. Thus, 5-HT3 receptors are responsible for activating guanylate cyclase in the hybrid cells. PMID- 2548884 TI - Identification of 5-HT2 receptors, alpha 1-adrenoceptors and amine release sites in rat brain by autoradiography with [125I]7-amino-8-iodo-ketanserin. AB - Using autoradiographical techniques, we found that [125I]7-amino-8-iodo ketanserin ([125I]AMIK) labelled with high affinity 5-HT2 receptors, alpha 1 adrenoceptors and sites involved in the release of biogenic amines and metabolites. The three binding sites for [125I]AMIK were identified by using selective inhibitors, i.e. BW 501 for 5-HT2 receptors, prazosin for alpha 1 adrenoceptors and tetrabenazine for the sites involved in the release of biogenic amines and metabolites. Using quantitative image analysis, we calculated the inhibition curves for the drugs in areas containing one of the receptor binding sites, i.e. the claustrum for 5-HT2 receptors, the thalamic nuclei for the alpha 1-adrenoceptors and the rostral part of the caudate putamen for the release sites. By using appropriate combinations of [125I]AMIK and drugs to occlude other binding sites, we could selectively label 5-HT2 receptors (in the presence of 0.1 microM prazosin and 1 microM tetrabenazine), alpha 1-adrenoceptors (in the presence of 1 microM BW 501 and 1 microM tetrabenazine) and release sites (in the presence 0.1 microM prazosin and 1 microM BW 501). Most binding to 5-HT2 receptors was observed in the cortical areas, the caudal caudate-putamen and the substantia nigra, while moderate binding was noticed in the nucleus accumbens, the olfactory bulb and the olfactory tubercle. There were many alpha 1 adrenoceptors in the numerous thalamic nuclei and few in the cortical areas, the periaqueductal grey matter and the substantia nigra.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548885 TI - K+ activity and regulation of intracellular pH in the sea urchin egg during fertilization. AB - Fertilization of the sea urchin egg is accompanied by changes in intracellular ion activities and transmembrane fluxes, which regulate the sequence of biochemical events of metabolic derepression. Changes in intracellular K+ activity during fertilization have been controversial and here we report our measurements using intracellular K+-sensitive microelectrodes. A small, but statistically significant, transient rise in internal K+ activity was detected during the first 10 min of fertilization. Since this change in K+ activity was ouabain sensitive, intracellular K+ activity in the fertilized egg appears to be regulated by the increased Na+, K+ ATPase activity, rather than the previously suggested K+ decompartmentalization. Increasing external K+ concentration was found to stimulate ouabain-sensitive alkalinization in the fertilized egg. The data are consistent with the possibility that Na+, K+ ATPase may regulate cytoplasmic pH by recycling Na+ that enters the cell through Na+-H+ antiport. PMID- 2548886 TI - The effects of bestatin, a microbial aminopeptidase inhibitor, on epidermal growth factor-induced DNA synthesis and cell division in primary cultured hepatocytes of rats. AB - We investigated the effects of microbial protease inhibitors, in particular the aminopeptidase inhibitor bestatin, on DNA synthesis and cell division induced by epidermal growth factor (EGF) in hepatocytes. Although bestatin did not significantly affect binding of EGF to hepatocytes, it inhibited EGF-induced DNA synthesis and cell division. DNA synthesis in rat hepatocytes was maximal 24-26 h after EGF addition to the medium. The time required for maximal DNA synthesis was not affected if bestatin was removed less than 12 h after addition, but synthesis was partially inhibited if bestatin was added to the medium several hours after EGF addition, depending on the time of bestatin addition. Our results suggest that bestatin arrests the new cell cycle induced by EGF at about 12 h after the initiation. Considering also our results obtained by employing other protease inhibitors, we concluded that specific proteases play important roles in hepatocyte DNA synthesis and cell division induced by EGF. PMID- 2548887 TI - Instability of extrachromosomal cosmid DNA in SV40-transformed human (ataxia telangiectasia) cells. AB - The ability of SV40-transformed human (ataxia-telangiectasia) fibroblasts to maintain Epstein-Barr virus (EBV)-based plasmids and cosmids extrachromosomally has been investigated. Transfection of a culture of cells with two different plasmids gave rise to cell clones which were able to maintain both plasmids extrachromosomally. When an EBV-based cosmid library was transfected into the cells and an individual cell clone was isolated, the extrachromosomal DNA derived from the cosmid contained numerous deletions and rearrangements. When individual cosmids were transfected into the culture, and several cell clones were isolated, the intracellular cosmid-derived DNA again showed the presence of multiple deletions and rearrangements. We conclude that although SV40-transformed cells are able to maintain more than one different EBV-based plasmid extrachromosomally, large EBV-derived molecules are extensively rearranged. SV40 transformed human fibroblasts cannot therefore be usefully used in attempting to clone genes from EBV-based cosmid libraries. PMID- 2548888 TI - Loss of nerve growth factor receptor-containing neurons in Alzheimer's disease: a quantitative analysis across subregions of the basal forebrain. AB - Magnocellular neurons comprising the Ch1-Ch4 regions of the basal forebrain provide topographic cholinergic innervation to the cerebral cortex, thalamus, and basolateral nucleus of the amygdala. Most quantitative studies analyzing the status of these neurons in Alzheimer's disease (AD) have employed Nissl-stained preparations. These studies principally analyzed large neurons of a prespecified cell diameter. Since basal forebrain neurons atrophy in Alzheimer's disease, an immunocytochemical marker for these neurons would appear to be a better alternative for determining whether there is regionally specific degeneration of cholinergic neurons across subregions of the basal forebrain. Brain sections from seven AD and five aged-matched control patients were immunocytochemically stained with a monoclonal antibody raised against the receptor for nerve growth factor (NGF), a probe which has previously been demonstrated to extensively and exclusively colocalize with cholinergic basal forebrain neurons in humans (17, 25, 35). NGF receptor-immunoreactive neurons within the hippocampal projecting nuclei of the medial septum (Ch1) and vertical limb of the diagonal band (Ch2) were minimally affected in AD as compared to control cases. In contrast, the Ch4 region demonstrated a significant loss of NGF receptor-immunoreactive neurons in AD that inversely correlated (-0.786) with the duration of the disease process. All four subregions of Ch4 were affected in the AD cases with the anterolateral (76.4%), intermediate (62.1%) and posterior divisions (76.5%) demonstrating the greatest reduction in NGF receptor-immunoreactive neurons. Nissl-counterstained sections failed to reveal magnocellular neurons which were not immunoreactive for the NGF receptor, suggesting that reductions in immunocytochemically stained neurons reflects neuron loss and not the failure of viable neurons to synthesize NGF receptors. These data indicate that cholinergic basal forebrain neurons which project to the amygdala, as well as to the temporal, frontobasal, and frontodorsal cortices, are most affected in AD. PMID- 2548889 TI - Plasticity-related binding of GABA and muscarinic receptor sites in piriform cortex of rat: an autoradiographic study. AB - This study has used the recently developed in vitro quantitative autoradiographic technique to examine the effects of olfactory bulb (OB) removal on receptor binding sites in the deafferented piriform cortex (PC) of the rat. The gamma aminobutyric acid-benzodiazepine receptor (GABA-BZR)- and muscarinic cholinergic receptor (MChR)-binding sites in layer I of PC were localized using [3H]flunitrazepam and [3H]quinuclidinyl benzilate as ligands, respectively. From the resultant autoradiograms the optical densities were measured using a Drexel DUMAS image analysis system. The densities of BZR and MChR-binding sites were markedly increased in the PC ipsilateral to the lesion as compared to the contralateral side in those subjects that were operated in adulthood (Postnatal Day 100, PN 100). Comparisons between the unoperated and PN 100 operated animals also showed significant increases in the deafferented PC. In the animals operated on the day of birth (PN 0) no significant differences were seen between the operated and the contralateral PC. The difference between the PN 0 deafferented PC and the unoperated controls shows a slight decrease in BZR density in the former group; however, in case of the MChR there is a slight increase on the side of the lesion. These results demonstrate that deafferentation of PC by OB removal appears to modulate both the BZR-binding sites that are coupled with the GABA-A receptor complex and the MChR-binding sites. The results also suggest that possibility of a role for these neurotransmitter receptor-binding sites in plasticity following deafferentation. PMID- 2548890 TI - Beta-adrenergic receptor binding in brain of alcoholics. AB - The binding of agonists and antagonists to beta-adrenergic receptors in brain tissue obtained postmortem in nonalcoholic controls and matched intoxicated and sober alcoholics was measured to assess the state of the receptors and their coupling to adenylate cyclase. Binding of antagonist, iodocyanopindolol, to cerebral cortical and cerebellar membrane preparations was not different in alcoholics compared to that in controls, suggesting that the number of beta adrenergic receptors was not affected by chronic ethanol ingestion. Agonist binding data, however, indicated the loss of the high-affinity agonist binding state of the beta-adrenergic receptor, representing the receptor-guanine nucleotide binding protein (Gs) complex. Such changes were observed in cerebral cortex but not in cerebellum of intoxicated alcoholics. These data suggest that cerebral cortical beta-adrenergic receptors are uncoupled from adenylate cyclase in these subjects. In cerebral cortical and cerebellar membranes of sober alcoholics both the high- and low-affinity agonist binding sites were observed. These findings are similar to those seen in animal studies and suggest that the effect of chronic ethanol ingestion on beta-adrenergic receptor-adenylate cyclase coupling is brain region specific and reversible with abstinence. Ethanol-induced changes in the coupling of receptors to adenylate cyclase may contribute to the physiological and behavioral manifestations of alcohol abuse. PMID- 2548892 TI - Dual granule localization of the dormant NADPH oxidase and cytochrome b559 in human neutrophils. AB - The subcellular localization of the microbicidal nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and associated b-cytochrome was investigated in human neutrophils. In unperturbed neutrophils 85% of b-cytochrome and the major part of membrane-bound components of the NADPH oxidase co sedimented with markers for specific granules and gelatinase. Using cytochrome b559 as a marker for membrane-bound components of the NADPH oxidase in quantitative studies we observed that, of the remaining 15%, the vast majority co sedimented with latent alkaline phosphatase, a marker for a newly identified mobilizable intracellular compartment. Only a small fraction co-localized with the plasma membranes. Azurophil granules contained a protease activity which rapidly inactivated the NADPH oxidase components present in other membranes. Stimulation of the neutrophils with formyl-methionyl-leucyl-phenyl-alanine and leukotriene B4 which caused minimal degranulation of specific granules, resulted in translocation of b-cytochrome to the plasma membrane, concomitant with incorporation of alkaline phosphatase into the plasma membrane. PMID- 2548893 TI - Eleven cases with lowered eosinophil peroxidase activity. PMID- 2548891 TI - Immune status towards Epstein-Barr virus in a group of Sicilian children. AB - The prevalence of antibodies to Epstein-Barr virus-determined antigens was studied in 17 children with acute infectious mononucleosis (IM) and in 263 children hospitalized for diseases unrelated to EBV infection. Antibodies against Epstein-Barr viral capsid antigens (VCA) were observed in 173 patients of the control group (66%), but 58 of them (33,5%) had not yet developed antibodies against Epstein-Barr virus-associated nuclear antigen (EBNA). IgM-specific antibodies were not found in any of the children of the control group but were present in all of the 17 patients with IM. The rates of positivity for IgA anti VCA and IgG anti-early antigen (EA) were similar in all age groups. Anti-viral capsid antigen IgG seropositivity increased to 83% by the age of six years, the mean geometric titre being highest between the ages of 2 and 4 years. Our results suggest that in Sicilian children the primary infection occurs prevalently early in life, in parallel with the occurrence of IM. PMID- 2548894 TI - [The participation of brain mediator and peptidergic systems in the mechanisms of the conditioned reflex]. AB - In dogs with implanted chemotrodes and microelectrodes, the role of acetylcholine , dopamine-, GABA- and P-ergic systems of the caudate nucleus and amygdala in positive and inhibitory conditioning, was studied. Activation of the same transmitter system in different structures could exert different behavioral effects, whereas activation of different transmitter subcortical systems could induce the same behavioral responses. The effects of activating or blocking agents upon the mediator subcortical system depended on the functional state of the nervous-system at the moment of administration as well as on the site of administration. It seems difficult to predict a contribution of one or another subcortical structure in organization of behavioral acts. PMID- 2548895 TI - [Comparison of the effects of thyroliberin and ACTH4-7 PGP on the learning of rats during the solving of spatial orientation tasks]. AB - The influence of TRH (100 micrograms/kg) and ACTH4-7 Pro-Gly-Pro (15 micrograms/kg and 25 micrograms/kg) involved an acceleration of the spatial learning on the 12-arm radial maze and on T-maze in rats; an increase in the number of correct choice after short (5-10 min) retention interval; and affected the working and reference spatial memory in the test of reinforced place performance. The effects of the TRH and ACTH4-7 Pro-Gly-Pro did not interfere with one another. PMID- 2548896 TI - [The brain bombesinergic system]. PMID- 2548897 TI - [Effect of vasopressin on the osmotic permeability of the bladder wall in the frog and its content of cAMP, cGMP and inositol triphosphate]. AB - An increase in the water permeability of the frog urinary bladder due to vasopressin, correlates with an increase of cAMP content in the bladder tissue. The osmotic permeability reached its maximum in 15-20 min. The sharp increase of inositol triphosphate content was observed within 20 sec after vasopressin administration, whereas cGMP content significantly decreased within 5 min. The augmentation of cAMP content seems to lead to a rise in water permeability white inositol triphosphate and cGMP acted, probably, as modulators of the vasopressin effect. PMID- 2548898 TI - [The participation of met-enkephalin in the display of contractile reactions of the large intestine and ileocecal sphincter]. AB - Blockade of H-cholinoreceptors, alpha- and beta-adrenoreceptors did not lead to disappearance of the colon and ileocecal sphincter contractile responses to i.a. administration of met-enkephalin (0.05-8.0 micrograms). The responses of the colon, as well as ileum and jejunum were augmented under blockade of alpha adrenoreceptors whereas the responses of ileocecal sphincter decreased. The latters were augmented under blockade of beta-adrenoreceptors. Met-enkephalin infused into the vascular bed of an isolated portion of the intestine (0.01-5.0 micrograms/ml/min), potentiated the colon and ileocecal sphincter responses to acetylcholine (1.0-20.0 micrograms), the colon responses to stimulation of the n. vagus efferent fibers, and the ileocecal sphincter responses to serotonin (20-100 micrograms). PMID- 2548899 TI - Vitamin D3 in dermatology: a critical appraisal. AB - Vitamin D3 and its active metabolites can be generated within the skin. Their physiological activities encompass the regulation of calcium homeostasis, protooncogene expression and the production of a number of intracellular and secretory molecules from cells in various organs, including keratinocytes, fibroblasts and leukocytes. The major outcome of these activities is a decreased cellular proliferation and the modulation of inflammation and immunity, suggesting that these agents might be effective in the treatment of psoriasis, ichthyosis and allergic dermatitis. Several open and double-blind studies have indeed demonstrated some efficacy in psoriasis, but healing is slow and often incomplete. Higher doses are more effective but increase the potential of calcium deposition in the vascular tree and particularly the kidneys. Clinical results in ichthyosis and allergic dermatitis are disappointing. For the future, therapeutically more effective analogues of vitamin D3 with no calcium liability need to be developed. PMID- 2548900 TI - Primary anaplastic large cell lymphoma of the skin. AB - Anaplastic large cell lymphoma is a subtype of nodal and extranodal lymphoproliferative disorder characterized by peculiar histopathological features and the positivity of lymphoid proliferating cells for Ki-1/Ber-H2 monoclonal antibody. A case, misdiagnosed as cutaneous metastasis and treated by surgery, in a woman with a history of previous neoplasms, is reported. In spite of the histopathological malignancy of the tumor, the patient is alive and free from recurrent or metastatic disease after 3 years of follow-up. PMID- 2548901 TI - Paget's disease of the nipple resembling an acantholytic disease on microscopic examination. AB - Two biopsies of an erosive lesion of the nipple had an appearance of an acantholytic disease without showing malignant cells. Only a third biopsy through the nipple with removal of a larger portion revealed some nests of atypical, large cells with clear cytoplasm, typical of Paget's disease. Immunohistochemical findings with carcinoembryonic antigen confirmed the diagnosis of Paget's disease of the nipple. This is the first case of Paget's disease which shows extensive acantholysis on microscopic examination and which resembles pemphigus vulgaris histologically. Acantholytic diseases are easily distinguished from Paget's disease and have never been mentioned in the differential diagnosis of this disease. A large biopsy through the nipple with the removal of a liberal portion of the nipple is suggested in every case of a suspected unilateral lesion of the nipple in order to avoid the overlooking of small nests of Paget's cells, as in our first biopsies, showing a histological picture of an acantholytic disease. PMID- 2548902 TI - Liver plasma membrane domains and endocytic trafficking. PMID- 2548903 TI - Mitogenic signalling in murine 3T3 cells: the cyclic AMP pathway. PMID- 2548904 TI - Lipoprotein receptors and atherosclerosis. AB - The whole lipoprotein spectrum of human plasma may be divided into atherosclerotic and anti-atherosclerotic lipoproteins. To the first class belong apolipoprotein (apo) B and some apoE-containing lipoproteins of the very-low density (VLDL), intermediate-density (IDL) and low-density (LDL) lipoprotein fractions. Anti-atherosclerotic lipoproteins are apoA-containing high-density lipoproteins (HDL). Circulating plasma lipoproteins are catabolized mainly by specific cell surface receptors (R) which react with apoB and apoE (B/E-R), for apoE (E-R) or for apoA (HDL-R). Whereas the B/E-R and E-R are responsible for the cellular uptake of lipoproteins and their lipid load by various organs, HDL-R are thought to promote lipid (cholesterol) efflux. There is an additional class of lipoprotein receptors, the so called scavenger-R which are responsible for the removal of altered or degraded lipoproteins for the circulation. Under normal physiological conditions, the concerted action of these receptors warrants efficient lipoprotein turnover and direction into target organs. Derangements of this system, however, may lead to the deposition and accumulation of atherogenic lipids, notably free cholesterol (FC) and cholesteryl esters (CE) in arterial tissue causing atherosclerosis and cardiac death. PMID- 2548905 TI - Vitamin D and calcitonin treatment in patients with femoral neck fracture: a prospective controlled clinical study. AB - The effects on general and bone metabolism of femoral neck fracture patients of 0.25 micrograms alpha-calcoid given orally twice daily (n = 9) and 25 micrograms calcitonin given subcutaneously 30 times (n = 10) in 10 weeks were studied against a control (n = 11). Bone histology and histomorphometry showed non-age related osteoporosis in 30% and osteomalacia in 22% of the patients studied. Impaired serum vitamin D status was found in 47-88% of patients, secondary hyperparathyroidism and increased serum parathyroid hormone in 59% and decreased serum calcitonin levels in 69%. On histology, normal findings and non-age related osteoporosis on histology were associated with low serum levels of 25 hydroxyvitamin D3, 1,25- and 24,25-dihydroxyvitamin D3. Very high serum levels of 1,25-dihydroxyvitamin D3 and low levels of 25-hydroxyvitamin D3 occurred in fracture patients with osteomalacia. Calcitonin improved calcium balance, reduced osteoporosis and increased the serum 1,25- and 24,25-dihydroxyvitamin D3 levels but had no effect on osteomalacia. Vitamin D reduced osteomalacia, slightly increased the serum 1,25-dihydroxyvitamin D3 concentration and decreased serum levels of parathyroid hormone. Both treatments gave a similar slight decrease in serum calcitonin concentrations. A mechanism of action for the treatments is suggested. PMID- 2548906 TI - Cellular retinoic acid-binding protein and the role of retinoic acid in the development of the chick embryo. AB - The distribution of cellular retinoic acid-binding protein (CRABP) in four stages of chick development is described using an affinity-purified antibody against rat CRABP. CRABP is the protein to which retinoic acid (RA) binds when it enters cells and may reflect the requirement of those cells for RA. We found several discrete cell populations which showed high levels of immunoreactivity. Some were in the neural tube such as the commissural neurons and the dorsal roof plate. Some were of neural crest origin such as the dorsal root ganglia, sensory axons, sympathetic ganglia, and enteric ganglia. The remaining populations were certain connective tissue cells, limb bud cells, and the myotome. These results suggest that certain organ systems, particularly the nervous system, have a requirement for RA during development and they may further our understanding of the teratogenic effects of retinoids on the embryo. PMID- 2548907 TI - Growth cone-growth cone interactions in cultures of rat sympathetic neurons. AB - Growth cones of sympathetic neurons from the superior cervical ganglia of neonatal rats were studied using video-microscopy to determine events following contact between growth cones and other cell surfaces, including other growth cones and neurites. A variety of behaviors were observed to occur upon contact between growth cones. Most commonly, one growth cone would collapse and subsequently retract upon establishing filopodial contact with the growth cone of another sympathetic neuron. Contacts resulting in collapse and retraction were often accompanied by a rapid and transient burst of lamellipodial activity along the neurite 30-50 microns proximal to the retracting growth cone. In no instances did interactions between growth cones and either fibroblasts or red blood cells result in the growth cone collapsing, suggesting that a specific recognition event was involved. On several occasions, growth cones were seen to track other growth cones, although fasciculation was rare. In some cases, there was no obvious response between contacting growth cones. Growth cone-growth cone contact was almost four times more likely to result in collapse and retraction than was growth cone-neurite contact (45% vs 12%, respectively). These observations suggest that the superior cervical ganglion may be composed of neurons with different cell surface determinants and that these determinants are more concentrated on the surface of growth cones than on neurites. These results further suggest that contact-mediated inhibition of growth cone locomotion may play an important role in growth cone guidance. PMID- 2548908 TI - Decreased platelet phosphoinositide turnover and enhanced platelet activation in IDDM. AB - Individuals with diabetes mellitus may have increased in vivo platelet activity. Abnormal platelet function could contribute to the increased incidence of vascular disease in diabetes mellitus. The biochemical mechanism(s) for platelet hyperactivation is unknown. We examined the hypothesis that platelet phosphoinositide turnover, a key signal-transducing mechanism involved in platelet activation, was abnormal in diabetic subjects. Platelets were harvested from 16 subjects with insulin-dependent diabetes mellitus (IDDM) and 19 healthy, nondiabetic control subjects of comparable age. Plasma beta-thromboglobulin (beta TBG), a specific marker of platelet activity in vivo, was increased in IDDM (67.1 +/- 7.3 ng/ml) compared with control (41.0 +/- 6.0 ng/ml) subjects (P less than .005). [32P]orthophosphate (32Pi) incorporation into the individual phosphoinositides and phosphatidic acid (PA) reached isotopic equilibrium by 120 min for IDDM and control subjects. Specific activity (dpm 32P/micrograms phosphorus) of phosphatidylinositol 4-phosphate (PIP) and phosphatidylinositol 4,5-bisphosphate (PIP2) was not different between IDDM and control subjects. Under these conditions, basal 32Pi incorporation into PIP2 and PIP but not phosphatidylinositol (PI) or PA was significantly lower in IDDM subjects. There was significantly decreased [32P]PIP2 and [32P]PIP hydrolysis and decreased [32P]PA formation in IDDM after platelet stimulation with 4 U/ml human thrombin. There were no differences in [32P]PI hydrolysis between the two groups. The mass of PIP2 was reduced (P less than .005) in the platelets from IDDM (0.71 +/- 0.23 nmol/10(9) platelets) compared with control (1.65 +/- 0.53 nmol/10(9) platelets) subjects. Similarly, PIP was lower (P less than .001) in IDDM (0.66 +/- 0.09 nmol/10(9) platelets) than in control (2.92 +/- 0.43 nmol/10(9) platelets) subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2548909 TI - Effects of phosphatidic acid on islet cell phosphoinositide hydrolysis, Ca2+, and adenylate cyclase. AB - Phosphatidic acid may be raised in glucose-stimulated islet cells through hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) and de novo synthesis with glucose-derived trioses. The mechanism by which exogenous phosphatidic acid from egg yolk lecithin may augment insulin secretion was investigated in neonatal beta-cells. In whole cells labeled with [2,8-3H]-adenine, a dose-dependent increase in phosphatidic acid-stimulated adenylate cyclase activity was seen, and a small intracellular transient free-Ca2+ rise was seen in Fura 2AM-loaded cells. In [gamma-32P]ATP-labeled membranes from those beta-cells, phosphatidic acid effected PIP2 hydrolysis. These phosphatidic acid-stimulated effects were not sensitive to preincubation with Bordetella pertussis exotoxin. The findings are consistent with a stimulatory effect of exogenous phosphatidic acid on insulin release and indicate an effect at the plasma membrane. It is possible that newly synthesized phosphatidic acid may function similarly to amplify intracellular events in glucose-stimulated islet cells through both local Ca2+ concentration and cyclic AMP-sensitive mechanisms. The participation of newly synthesized phosphatidic acid derived from glucose could provide a link between the metabolism of glucose and insulin release. PMID- 2548910 TI - Production of endothelin 1 by cultured bovine retinal endothelial cells and presence of endothelin receptors on associated pericytes. AB - Endothelinlike immunoreactivity was detected by radioimmunoassay in medium conditioned by cultured endothelial cells obtained from bovine retinal microvessels (9.2 +/- 6.5 pM, n = 4). Sephadex G-25 column chromatography and fast-protein liquid chromatography revealed that most of the endothelinlike immunoreactivity was eluted in an identical position to synthetic endothelin 1. Retinal capillary pericyte-conditioned medium contained 2.9 pM endothelinlike immunoreactivity. In contrast to endothelial cells, retinal pericytes were found to bind endothelin. The dissociation constant and binding capacity were 0.14 nM and 1.5 x 10(5) sites/cell (n = 3), respectively. These findings suggest that endothelin produced by the retinal endothelial cells binds to the pericytes, adding support to the suggestion that pericytes in the retina may have a musclelike function. PMID- 2548911 TI - Octreotide (SMS 201-995) in the treatment of metastatic glucagonoma: report of one case and review of the literature. AB - We report 1 patient with a necrolytic migratory erythema, a high plasma glucagon concentration and a metastatic pancreatic endocrine tumor who has now been treated effectively for 33 months with the somatostatin analogue octreotide (SMS 201-995) (400 micrograms/day). The results of SMS 201-995 in the treatment of glucagonoma syndrome are reviewed. PMID- 2548912 TI - Gastrostomy-biliary drainage in a patient with bile duct cancer: a basis for multimodality treatment. PMID- 2548914 TI - Na+/H+ exchange in granulosa cells of the three largest preovulatory follicles of the domestic hen. AB - Sodium-dependent intracellular pH (pHi) regulation was compared in granulosa cells from the three largest avian ovarian follicles by monitoring the pHi with biscarboxyethylcarboxyfluorescein, a dye whose fluorescence increases with alkalinity. Collagenase-dispersed granulosa cells obtained from the largest (F1), second largest (F2), and third largest (F3) preovulatory follicles about 2-3 hr prior to expected ovulation of F1 were used in the present study. The resting pHi measured in nominally bicarbonate free buffer with extra-cellular Na+ (Nao+ = 144 mM) and external pH (pHo) of 7.3 was about 6.8 in cells from F1, F2, and F3. There was no correlation between the stage of follicular development and the pHi whether the follicles were removed in the early or late preovulatory period. After acute cytoplasmic acidification by exposure of cells to nigericin in choline+ buffer, or by the abrupt removal of ammonium chloride, complete recovery of pHi occurred in 4-5 min. The rate and magnitude of the recovery were dependent upon the concentration of Nao+ and were abolished when Nao+ was replaced completely by choline+. Recovery in the presence of Nao+ was inhibited dose dependently by amiloride (sodium-hydrogen exchange inhibitor). There was no difference between the rate and the extent of pHi recovery in acid-loaded cells obtained from F1, F2, and F3. Furthermore, by varying the concentration of Nao+ between 0 and 144 mM both young and matured granulosa cells extruded acid at the same rate. In addition, amiloride inhibited the Nao+ dependency of pHi recovery to a similar degree in F1, F2, and F3 cells. Our observations demonstrate in avian granulosa cells the existence of a Nao+-dependent, amiloride-sensitive pHi regulatory system that is equally effective in cells obtained from the three largest yolk-filled follicles. PMID- 2548913 TI - The presence of corticotropin-like and opiate-like activities in tissues of adult sea lamprey, Petromyzon marinus L. AB - Sexually mature landlocked sea lamprey were captured during their upstream migration. Different tissues, including the brain, pituitary, heart, liver, gut, testis, and ovary, were dissected from the animals and an acetone powder was prepared from each tissue. The tissue acetone powders were subjected to heat treatment and were then extracted with an acidic medium in order to inactivate any proteases present. The resulting acid acetone powders were then tested for their ability to stimulate corticosterone production from isolated rat adrenal cells and to displace the binding of D-Ala2-D-Leu5-[tyrosyl-3,5-3H]-enkephalin to rat brain membranes. It was found that the brain and liver contained steroidogenic activity while opiate activity was detected in the heart, liver, gut, brain, and pituitary. No steroidogenic activity was found in the heart, ovary, testis, gut, and pituitary while ovary and testis did not contain assayable opiate activity. None of the tissues contained beta-endorphin-like immunoreactivity. PMID- 2548916 TI - Early and delayed changes in potassium transport during the initiation of cell proliferation in CHO culture. AB - Stimulation by serum of cell proliferation in G1-arrested culture of Chinese hamster ovary cells CHO-K1 was accompanied by an early (during the first minutes) and delayed (2-10 h) activation of Na+,K+-ATPase and an increase in cell K+ content from 0.5-0.6 to 0.7-0.8 mmol per gram protein. Isoproterenol acted synergistically with serum in eliciting both early and delayed changes in K+ transport and in stimulating G1----S transition. Isoproterenol alone (without serum) induced a transient increase in K+ influx via Na+,K+-ATPase without changing the cell K+ content or having any mitogenic effect. Theophylline enhanced the serum-induced early activation of Na+,K+-ATPase but inhibited both the delayed increase in cell K+ and the G1----S transition. Early serum-induced increase in K+ transport was not affected by cycloheximide, whereas net accumulation of cell K+ was abolished by the drug. It is concluded that the early and the delayed activation of Na+,K+-ATPase induced by mitogens can be dissociated; the early ionic response is related to the primary transduction of membrane signal, whereas the delayed modulation of ion transport via Na+,K+ ATPase has another function and is associated with cell growth. PMID- 2548915 TI - Amphiphilic derivatives of betaine esters as modifiers of macrovesicular BLM. AB - A series of amphiphilic derivatives of betaine esters (V-n), with the chemical structure (CH3)3N+COOCnH2n + 1Cl- (n = 10, 12, 14 or 16) were studied with respect to their effects on the electrical properties of lecithin macrovesicular membranes. Normalized resistance and breakdown voltage were found to depend on the V-n concentration in the membrane and on the alkyl chain length (n). Resistance decreases up to about 10(4) ohm.cm2 and breakdown voltage decreases by 111 mV were detected in the V-n: lecithin molar ratio range measured (0.005 0.05). Maximal decrease in breakdown voltage was observed for V-14. These findings together with the featured anionic selectivity suggest that, due to the interaction of V-n with phospholipids, hydrophilic pores are formed in the lipid bilayers. This assumption is supported by the results obtained by electron paramagnetic resonance (EPR) measurements which showed no collective changes in bilayer dynamics or ordering. In particular, rotational correlation times and order parameters of the spin probe molecules dissolved in the membrane did not change in the concentration range tested. Since a large number of defects in the membrane can be expected to influence the collective ordering and dynamics, this observation also suggest that the number of pores formed is small. PMID- 2548917 TI - Hypothetical structure of the ATP-binding site of (Na+ + K+)-ATPase. PMID- 2548918 TI - Laminar distribution of benzodiazepine receptors in visual cortex of adult rat. AB - The characteristics and distribution of benzodiazepine receptors in individual layers of the visual cortex of adult rats were examined with the 3H-flunitrazepam binding technique employed on intact tissue slices. The different visual cortical layers were separated by cutting serial cryocut sections horizontally to the cortical surface and collecting the slices from each individual cortical layer under anatomical control. Highest benzodiazepine receptor densities were found in layers IV and VI. A moderate receptor density was detected in layer V (80% of highest density). The lowest receptor binding was observed in cortical layers I and II/III, still representing 66% of the highest receptor density. Binding affinities varied slightly between layers with dissociation constants somewhat higher for layers IV to VI in comparison to layers I and II/III. The distinct laminar pattern of benzodiazepine receptors in rat visual cortex suggests a differential neuromodulatory significance of these receptors in each individual cortical layer. PMID- 2548919 TI - Spin label and microcalorimetric studies of the interaction of DNA with unilamellar phosphatidylcholine liposomes. AB - High-molecular DNA from chicken erythrocytes interacts with 1,2 dipalmitoylphosphatidylcholine in unilamellar liposomes, both in the presence and absence of Mg2+ ions. This interaction results in a phase separation in liposome membranes. The new phase induced by DNA and Mg2+ has a higher gel-liquid crystal phase transition temperature as measured by microcalorimetry. In the liquid crystalline state, the 16- and 5-doxyl stearic acid spin labels indicate changed local bilayer properties at the label position in the new phase. PMID- 2548920 TI - Genetic and physical mapping of the mcrA (rglA) and mcrB (rglB) loci of Escherichia coli K-12. AB - We have genetically analyzed, cloned and physically mapped the modified cytosine specific restriction determinants mcrA (rglA) and mcrB (rglB) of Escherichia coli K-12. The independently discovered Rgl and Mcr restriction systems are shown to be identical by three criteria: 1) mutants with the RglA- or RglB- phenotypes display the corresponding McrA- or McrB- phenotypes, and vice versa; 2) the gene(s) for RglA and McrA reside together at one locus, while gene(s) for RglB and McrB are coincident at a different locus; and 3) RglA+ and RglB+ recombinant clones complement for the corresponding Mcr-deficient lesions. The mcrA (rglA) gene(s) is on the excisable element e14, just clockwise of purB at 25 min. The mcrB (rglB) gene(s), at 99 min, is in a cluster of restriction functions that includes hsd and mrr, determinants of host-specific restriction (EcoK) and methyladenine-specific restriction respectively. Gene order is mcrB-hsdS-hsdM hsdR-mrr-serB. Possible models for the acqusition of these restriction determinants by enteric bacteria are discussed. PMID- 2548921 TI - The PET54 gene of Saccharomyces cerevisiae: characterization of a nuclear gene encoding a mitochondrial translational activator and subcellular localization of its product. AB - The product of the nuclear Saccharomyces cerevisiae gene PET54 is specifically required, along with at least two other nuclear gene products, for translation of the mitochondrial mRNA encoding subunit III of cytochrome c oxidase (coxIII). We have genetically mapped PET54 (to the right arm of chromosome VII, 4.8 cM centromere-distal to SUF15), and have biochemically characterized the gene and its product. We determined the nucleotide sequence of a 1.6-kb DNA fragment carrying PET54 and identified the PET54 reading frame by determining the sequence of an ochre mutant allele as well as frameshift and frameshift-revertant alleles of the gene. The wild-type PET54 gene encodes a slightly basic 293-amino acid protein. PET54 is expressed from two mRNAs, both with unusual features: a major transcript with an extremely short 5'-untranslated leader, and a minor transcript with a relatively long 5'-leader carrying three short open reading frames. Antiserum raised against a trpE-PET54 fusion protein was used to probe subcellular fractions. These experiments showed that the PET54 protein is specifically associated with mitochondria, suggesting that it is likely to act directly in coxIII translation. PMID- 2548923 TI - A recombinational hotspot at the triplo-lethal locus of Drosophila melanogaster. AB - In the genome of Drosophila melanogaster there is only one locus, Tpl, that is triplo-lethal; it is also haplo-lethal. Previous work has identified 3 hypomorphic alleles of Tpl which rescue animals carrying a duplication of Tpl, but which are not dominant lethals as null mutations or deficiencies would be. We have found that all three hypomorphic alleles act as site-specific hotspots for recombination when heterozygous with a wild-type homolog. Recombination between the flanking markers ri and Ki is increased 6.5-10.5-fold in the presence of Tpl hypomorphic alleles. The increased recombination was found to occur between Tpl and Ki, while recombination in other adjacent regions is unchanged. The use of isogenic Tpl+ controls, and the use of flanking intervals in the mutant chromosomes allows us to rule out the interchromosomal effect as a cause. We have also observed premeiotic recombination occurring at the Tpl hypomorphic alleles in male heterozygotes. We hypothesize that transposons are responsible for both the hypomorphic phenotype and the high frequency of recombination. PMID- 2548922 TI - Molecular evolution of the telomere-associated MAL loci of Saccharomyces. AB - The MAL gene family of Saccharomyces consists of five multigene complexes (MAL1, MAL2, MAL3, MAL4, and MAL6) each of which encodes maltose permease (GENE 1), maltase (GENE 2) and the trans-acting MAL-activator (GENE 3). Four of these loci have been mapped and each is located at or near the telomere of a different chromosome. We compare the physical structure of the MAL loci and their flanking sequences. The MAL loci were shown to be both structurally and functionally homologous throughout an approximately 9.0-kb region. The orientation of the MAL loci was determined to be: CENTROMERE . . . GENE 3-GENE 1-GENE 2 . . . TELOMERE. Telomere-adjacent sequences were found flanking GENE 2 of the MAL1, MAL3 and MAL6 loci. No common repeated elements were found on the centromere-proximal side of all the MAL1, loci. These results suggest that, during the evolution of this polygenic family, the MAL loci translocated to different chromosomes via a mechanism that involved the rearrangement(s) of chromosome termini. PMID- 2548924 TI - Phenotypic and molecular analysis of the facets, a group of intronic mutations at the Notch locus of Drosophila melanogaster which affect postembryonic development. AB - The function of the Notch locus of Drosophila melanogaster is essential for normal development both during embryogenesis and during postembryonic stages. In the embryo its function is necessary for the correct segregation of neural from epidermal lineages. During postembryonic stages Notch exhibits pleiotropic effects that are both tissue- and stage-specific. Here, we examine a group of six recessive mutations, the facets (fa, fa3, fag, fag-2, fafx and fasw), which affect eye morphology and have been previously shown to be associated with the insertion of transposable elements in an intronic region of Notch. The analysis of revertants has shown that the mutant phenotype depends on the presence of the transposable element and that the disruption of the wild-type sequence organization per se is not its cause. Four of these alleles, even though they are associated with the insertion of the same transposable element, display considerably different phenotypes. Therefore, no simple correlation exists between the mutant phenotype and the type of inserted element. A comparison of the tissue localization of the Notch and the transposable element transcripts revealed that in the third larval instar the elements are transcribed in both orientations in tissues in which Notch is also transcriptionally active. The complexity of the defects associated with the facet alleles, as well as the findings of the transcriptional analysis, indicate that a mutational mechanism based solely on transcriptional interference is not sufficient to explain the nature of the mutational event. It is likely that in these mutations alterations, in the temporal and/or spatial context caused by transcriptional and perhaps posttranscriptional interference mechanisms by the inserted elements, may be responsible for the mutant phenotype. PMID- 2548925 TI - The use of fish oil in bronchial asthma. AB - A fish-oil enriched diet has potential in modulating the humoral and inflammatory components of the allergic response by inhibiting the generation of pro inflammatory mediators derived from arachidonic acid and by reducing the production of PAF-acether. In addition, EPA suppresses the responses of target cells and tissues. Studies in bronchial asthma confirm the anti-inflammatory potential of a fish-oil enriched diet. Dietary supplementation with EPA in subjects with asthma led to changes in leukocyte mediator generation and chemotactic responses. There was also a significant attenuation of the late asthmatic response to inhaled antigen. Further studies are needed to determine the full potential of such diets in effecting changes in the clinical aspects of allergic disease. PMID- 2548926 TI - Cromolyn and nedocromil therapy of asthmatic children. AB - Controlled studies of cromolyn treatment of asthmatic children have revealed benefits beyond the incredible safety of the drug. It is now recognized that the patient with asthma will have decreased asthma and less bronchial hyperreactivity when treated for more than two months with cromolyn. Also, the reduced sensitivity of beta-receptors with chronic use of beta adrenergic drugs is abated with concomitant cromolyn use. It is hoped that the studies with nedocromil will reveal additional benefits with this class of antiasthmatic medication. PMID- 2548927 TI - Direct selection for curing and deletion of Rhizobium plasmids using transposons carrying the Bacillus subtilis sacB gene. AB - We have constructed derivatives of the transposon Tn5 carrying the mob site (oriT) of plasmid RP4, and an nptI-sacB-sacR cassette [Ried and Collmer, Gene 57 (1987) 239-246]. The mob site, in conjunction with the antibiotic-resistance markers carried on the transposons, allows identification of transposon inserts in cryptic plasmids by mobilisation to other strains. The sacB-sacR genes allow direct selection for the loss or curing of plasmids, because only strains which no longer contain an active sacB gene are able to grow on media containing sucrose. We have tested these transposons in four strains of Rhizobium leguminosarum and two strains of Rhizobium meliloti, and have been able to demonstrate curing of several large cryptic plasmids, and generation of large deletions in many other plasmids. This method has enabled us to show that the R. leguminosarum plasmids pRL12JI and pR1eVF39f carry auxotrophic markers, and that the plasmid pR1eVF39c carries genes which affect colony morphology. PMID- 2548928 TI - High-level synthesis of cowpea mosaic virus RNA polymerase and protease in Escherichia coli. AB - An expression system for the production of polymerase proteins of cowpea mosaic virus (CPMV) in Escherichia coli cells is described. High-level synthesis of proteins containing protease and polymerase moieties (110-kDa protein) and polymerase alone (87-kDa protein) were obtained from cells containing different plasmid constructions. Precursor and processed forms of CPMV proteins were detected by immunoblotting with antisera directed against 170-kDa precursor polyprotein and 24-kDa viral protease. Crude lysates and supernatant fractions of the lysates from E. coli cells harboring the various plasmid constructions were analysed for poly(A)-oligo(U) polymerase activity and found to be negative for CPMV activity under conditions where similar expression systems for the production of poliovirus RNA polymerase activity were positive. Thus, conditions for CPMV RNA replication may indeed be different from those for poliovirus even though the genomic organization of these viruses is similar. PMID- 2548929 TI - A simple procedure for transferring genes cloned in Escherichia coli vectors into other gram-negative bacteria: phenotypic analysis and mapping of TOL plasmid gene xylK. AB - A simple method to transfer non-conjugative Escherichia coli plasmids to other Gram-negative bacteria and their maintenance is described. This method involves generation of inverse transposition-mediated cointegrates of the non-conjugative E. coli plasmid with a conjugative IncW broad-host-range plasmid, R388, carrying Tn10. Isolation of such cointegrates was readily effected by conjugal transfer from an E. coli donor containing the two plasmids to an E. coli recipient, with selection for transconjugants expressing a marker of the E. coli plasmid. This method is particularly useful when large series of E. coli vector-based clones need to be expressed in other Gram-negative bacteria to be functionally analysed, either by complementation or recombination. Utility of the method is shown by a functional analysis in Pseudomonas putida of pBR322 hybrid plasmids containing catabolic genes of TOL plasmid pWW0. PMID- 2548931 TI - A new onset of fatigue in an active elderly man. AB - Fatigue is often perceived as part of normal aging. Yet for many active elderly, a complaint of generalized, non-specific weakness should alert the physician to the existence of possible underlying pathology, as this case demonstrates. PMID- 2548930 TI - Transposon-mediated restriction mapping of the Bacillus subtilis chromosome. AB - Analysis of chromosomal DNA depends upon a knowledge of the locations of restriction sites over several thousand kilobases (kb). However determination of even a subset of these sites can be time-consuming, and it can be difficult to link genetic and physical maps. We describe here a significant improvement which can be used in concert with genetically mapped chromosomal insertions. The circular chromosome of Bacillus subtilis 168 was physically examined on contour clamped homogeneous electric field (CHEF) gels using the restriction enzyme NotI. Restriction mapping of the 4.7-megabase (Mb) DNA was accomplished using a novel technique involving the transposon Tn917, which linked the genetic and physical maps and also significantly increased the rate at which this was performed. The DNA of 54 strains which contained Tn917 at genetically determined locations was cleaved with NotI and used to determine the approximate positions of 31 restriction fragments with sizes between 45 kb and 290 kb, totalling 3589 kb. This information should greatly assist in the construction of a more detailed map using standard methodology. PMID- 2548932 TI - [Gas-chromatographic method of determining the levels of tri-allate in the water, soil and plants]. PMID- 2548933 TI - Oesophageal symptoms, their causes, treatment, and prognosis in patients with the acquired immunodeficiency syndrome. AB - In a prospective study of 154 AIDS patients, 48 (31%) complained of pain on swallowing both liquids and solids and 32 (21%) of these also had dysphagia. While candidiasis was the most common cause of symptoms (26 patients), discrete ulceration of the oesophagus occurred in 12 instances in 10 patients (four cytomegalovirus, four herpes simplex virus, three aphthous ulcer, one peptic ulcer). One patient had a diffuse oesophagitis caused by Mycobacterium avium intracellulare. No cause was found for the oesophageal symptoms in four patients. Kaposi's sarcoma (KS) was found in seven patients associated with other pathology in four. All 26 patients with oesophageal candidiasis only, also had oral involvement. All the patients with herpes simplex virus (four) and aphthous ulcers (three) had obvious perioral involvement. Three of the four patients with cytomegalovirus ulceration had evidence of disease elsewhere (colon or retina). All patients with Kaposi's sarcoma lesions had skin and buccal cavity involvement. The cause of oesophageal disease was usually obvious at endoscopy. The appearance of candidiasis was typical and the various ulcerating lesions also had different macroscopic configurations. Cytomegalovirus infection produced deep linear ulcers in the distal oesophagus, herpes simplex oesophagitis is similar in appearance to the typical perioral lesions of fluid filled vesicles. Diagnostic radiology was not helpful in most patients. In nine of 17 patients with candidiasis, the barium swallow examination performed within 24 hours of presentation was normal. In only three of seven patients with oesophageal ulceration (three cytomegalovirus, two herpes simplex virus, one aphthous, one peptic) was there evidence of an abnormality. Treatment produces symptomatic relief. All patients with candidiasis responded to ketoconazole, the four with herpes simplex virus to acyclovir and one of three with aphthous ulceration had a dramatic response to thalidomide. The three patients with cytomegalovirus infection who were treated with foscarnet had a prolonged remission of symptoms. The overall prognosis of patients with oesophageal symptoms is poor, with an average survival time from a definitive diagnosis of five months (range one to 13). PMID- 2548934 TI - Human papillomavirus and epithelial ovarian neoplasia. AB - In contrast to the strong association between human papillomavirus (HPV) and squamous cell carcinoma of the lower female genital tract, no viral DNA had been found in epithelial ovarian carcinoma. Recently, however, R. H. Kaufman, J. Bornstein, A. N. Gordon, E. Adam, A. L. Kaplan, and K. Adler-Storthy [Gynecol. Oncol. 27, 340-349 (1987)] reported the detection of HPV-6 DNA by in situ hybridization in 10 of 12 patients with advanced epithelial ovarian carcinoma. To further investigate the possible association between HPV and epithelial ovarian neoplasia, tumor from 12 patients with epithelial ovarian adenocarcinoma, 3 with epithelial ovarian tumors of low malignant potential, and 3 with epithelial ovarian tumors of low malignant potential, and 3 with epithelial ovarian adenomas was examined for HPV DNA by the Southern hybridization technique. All the tissues were tested under low stringency for HPV-6 and under high stringency for HPV-6, HPV-16, HPV-18, HPV-31, and HPV-35. In addition, all tissues were tested by polymerase chain reaction for the presence of HPV-6 and HPV-11. Of the 12 patients with adenocarcinoma, 5 were poorly differentiated, 4 serous, 1 endometrioid, 1 mucinous, and 1 mixed. The neoplasms were of grades 2-4 with a predominance of stages III and IV (83%). Of the 3 patients with tumors of low malignant potential, all were serous and stage I. Of the 3 patients with adenomas, 1 was mucinous, 1 serous, and 1 Brenner. We were unable to detect HPV related sequences in any of the specimens. On the basis of these findings, there appears to be no association between HPV and epithelial ovarian neoplasia. PMID- 2548936 TI - Clinicopathologic analysis of uterine malignant mixed mullerian tumors. AB - A clinicopathologic evaluation of 60 patients (median age, 66 years) presenting for primary treatment of uterine malignant mixed mullerian tumors between 1959 and 1982 was conducted. Surgical stage was utilized for assessment of survival by stage. With a minimum follow-up of 5 years, overall 2- and 5-year Kaplan-Meier survival estimates were 53 and 39%, respectively; they were 75 and 58% when disease was confined to the uterus and 27 and 15% when disease extended beyond the uterus. By log-rank and logit-rank analysis, surgical stage and depth of invasion stratified by stage were significant prognostic determinants; no significant association was found with carcinoma grade, sarcoma mitotic figure count, sarcoma histologic subtype, sarcoma necrosis, or capillary-like space involvement. No significant survival advantage was found for surgery plus irradiation or surgery plus chemotherapy compared with surgery alone after stratification according to stage. Progression-free survival after complete extirpation of macroscopic disease was not significantly different, stage for stage, between surgery alone and surgery plus radiotherapy. PMID- 2548935 TI - Influence of secondary cytoreduction at the time of second-look laparotomy on the survival of patients with epithelial ovarian carcinoma. AB - The value of secondary cytoreductive surgery at the time of second-look laparotomy in patients with epithelial ovarian carcinoma is not established. Sixty-seven patients with residual carcinoma found at the time of second-look laparotomy performed at Memorial Sloan-Kettering Cancer Center between December 1, 1978, and May 30, 1986, were evaluated for survival relative to the success of secondary cytoreductive surgery. At second-look laparotomy, 17 patients had microscopic disease, 28 patients had disease less than 2 cm and 22 patients had disease greater than 2 cm. After secondary cytoreductive surgery 33 patients had microscopic disease, 26 patients had disease less than 2 cm, and 7 patients had disease greater than 2 cm (1 unknown). Five-year survival by Kaplan-Meier calculation was 62% for patients found to have microscopic disease at second-look laparotomy and 51% for patients whose disease was rendered microscopic by secondary cytoreductive surgery (P = 0.55). Patients left with gross disease (either less than or greater than 2 cm) had 5-year survivals of less than 10% (P = 0.013 compared with microscopic residual). Secondary cytoreductive surgery at the time of second-look laparotomy in patients with epithelial ovarian carcinoma may result in improved survival of patients who are reduced to microscopic residual disease. PMID- 2548937 TI - DNA content as a prognostic index in gestational trophoblastic neoplasia. AB - Hydatidiform mole will progress to malignant gestational trophoblastic neoplasia (GTN) in some cases. Aneuploidy and high proliferative activity are associated with malignant tumors. Molar pregnancy tissue was considered a precursor to malignant GTN, and was studied retrospectively using paraffin-embedded tissue to determine whether aneuploidy or proliferative rates measured on molar tissue could predict a malignant course. Tissues from 51 complete hydatidiform moles were analyzed for nuclear DNA content by flow cytometric techniques. A chart review identified the clinical course after evacuation of the mole. A satisfactory DNA histogram was generated in 40 cases. Of the 40 patients, 22 (55%) had spontaneous resolution, and 18 patients (45%) required treatment for persistent GTN. The molar tissue was found to be euploid in 27 cases and aneuploid in 13 cases. Eight of the twenty-seven euploid cases (30%) required treatment after evacuation, whereas 10 of the 13 aneuploid cases (77%) required treatment after molar evacuation. Proliferative index (PI) was compared with treatment requirements. Average PI was 0.11 +/- 0.10 for the treatment group and 0.08 +/- 0.06 for the spontaneous resolution group. The correlation of clinical course with ploidy was significant (P less than 0.01). The association with proliferative index was not (P greater than 0.05). Aneuploidy, therefore, identifies a high-risk group of molar pregnancies, and may represent those that have undergone one stage of malignant transformation. PMID- 2548938 TI - [Unusual course of malignant synovioma of the flexor tendon sheath of the hand]. AB - The authors report a synovial sarcoma of the hand with an unusual case history and a favorable outcome after two years. A forty-four-year old woman suffering from uncharacteristic pain in the right hand with slightly abnormal EMG-findings for the median nerve was treated by operative release of the carpal tunnel. No unusual findings were discovered intraoperatively. The patient continued suffering from disabling pain for another year so that a second operation was performed by another surgeon. He discovered a tumor behind the median nerve which could not be removed radically because of profuse bleeding. In spite of diagnosing a sarcoma, the patient refused further surgery, however combined radio and chemotherapy was administered for one year. The tumor was reduced in size but did not disappear. Following open biopsy and proved presence of sarcoma, the remaining tumor was removed by amputating the ulnar part of the hand. Two years following this last operation, the patient is free of local symptoms and there is no evidence of metastases. PMID- 2548939 TI - [No negligence in preventive vaccinations!]. PMID- 2548940 TI - Prevention of CCL4-induced liver cirrhosis by silymarin. AB - The efficacy of silymarin treatment in preventing biochemical and histological alterations in CCL4-induced liver cirrhosis in rats was studied. Four groups of rats were treated with: (1) CCL4; (2) mineral oil; (3) CCL4 + silymarin; and (4) silymarin. All animals were sacrificed 72 h after the end of treatments. The activities of alkaline phosphatase (alk. phosp.), gamma-glutamyl transpeptidase (GGTP), glutamic pyruvic transaminase (GPT) and glucose-6-phosphatase (G6Pase), and bilirubin content were determined in serum. Na+, K+-ATPase and Ca++-ATPase activities were measured in isolated plasma membranes. Lipoperoxidation, triglycerides (TG), and glycogen contents were also measured in liver homogenates. Liver cirrhosis was evidenced by significant increases in liver collagen, lipoperoxidation, serum activities of alk. phosp., GGTP, GPT, G6Pase, bilirubin content, and liver TG. Activities of ATPases determined in plasma membranes were significantly reduced, as was liver glycogen content. Silymarin cotreatment (50 mg/kg b.wt) completely prevented all the changes observed in CCL4 cirrhotic rats, except for liver collagen content which was reduced only 30% as compared to CCL4-cirrhotic rats. Silymarin protection can be attributed to the agent's antioxidant and membrane-stabilizing actions. PMID- 2548941 TI - Effect of dopamine, ibopamine, and epinine on alpha- and beta-adrenoceptors in canine pulmonary circulation. AB - Dopamine has been widely utilized in the treatment of acute congestive heart failure, ibopamine, the diisobutyrate ester of N-methyldopamine (epinine), is a novel inotropic agent that, unlike-dopamine, is orally active. In clinical studies at doses that produce favorable hemodynamic responses, ibopamine and dopamine can evoke a slight and transient increase in pulmonary artery pressure and pulmonary capillary wedge pressure, an effect that is no longer apparent 1 h after administration. We have previously demonstrated in anesthetized dogs that this effect is due to stimulation of alpha-adrenoceptors in the pulmonary circulation by dopamine and ibopamine, as well as by the active form of ibopamine, epinine. The aim of the present investigation was to determine how dopamine, ibopamine, and epinine interact with beta-adrenoceptors in the canine pulmonary circulation, since this activity may serve to offset the alpha adrenoceptor-mediated pulmonary vasoconstrictor responses. Intraarterial injection of dopamine, ibopamine, and epinine resulted in dose-dependent pulmonary vasoconstrictor responses with a maximum increase of approximately 50 60% above resting pulmonary vascular tone. When animals were pretreated with propranolol (1 mg/kg iv) to block beta-adrenoceptors, pulmonary vasoconstrictor responses to dopamine were unchanged, whereas pulmonary vasopressor responses to ibopamine and epinine were significantly potentiated, especially for epinine. Upon intraduodenal administration of a therapeutically effective dose of ibopamine (i.e. 36 mg/kg) to normal dogs, virtually no pulmonary pressor response was observed. However, administration of this same dose of ibopamine to dogs pretreated with propranolol (1 mg/kg iv) resulted in a marked pulmonary pressor response. These data indicate that epinine, and therefore the parent compound ibopamine, have the capacity to stimulate beta 2-adrenoceptors in the pulmonary circulation to a far greater degree than dopamine, and that this activity serves to offset, at least in part, the alpha-adrenoceptor-mediated pulmonary vasoconstriction that occurs in response to ibopamine. PMID- 2548942 TI - [Experimental studies on the mechanism of spinal cord ischemia--the state of free radical scavengers]. AB - I have developed the new procedure for the preparation of experimental ischemic spinal cord from dog. Using this preparation, I have measured the enzymatic activities of superoxide dismutase (SOD), catalase, and glutathione peroxidase. The ischemic spinal cords, monitored by the evoked spinal cord potential, were obtained after clamping bilateral subclavian arteries and aorta distal to the origin of the left subclavian artery. Total SOD activity of the normal spinal cord was one ninth lower than that of the brain. But its catalase activity was eight times higher. Total SOD, Mn-SOD, and glutathione peroxidase activities of my experimental ischemic spinal cord didn't change as compared with those of the normal ones. The catalase activity was decreased after spinal cord ischemia. These results indicate that the catalase plays the main of protection against peroxidative damage in the spinal cord induced by reperfusion, Ca++-dependent protease system may be involved in the decrease for catalase activity in addition to the high turnover rate of this enzyme. In summary, ischemic injury of spinal cord could be induced from the active oxygen species as well as disorder of Ca++ in the tissue. PMID- 2548943 TI - [The study on in vitro transformation by human papillomavirus type 16 E6/E7 region]. AB - A NIH3T3-derived cell clone (NA7) in which human papillomavirus (HPV) 16 early region E6/E7 was inducible by dexamethasone (DXM) under the control of mouse mammary tumor virus long terminal repeat was established. A transforming function for HPV 16 E6/E7 region was analyzed by this established clone. Northern blot hybridization demonstrated that the increased expression of PCNA/cyclin and c-myc at the confluent state in accordance with the induced expression of E6/E7 region by DXM. Although complete transformation was not observed, the saturation density of NA7 was increased by the addition of DXM. The transfection assay using NA7 showed that adenovirus type 5 E1B (Ad5E1B) could cooperate with E6/E7. Furthermore, it was indicated that E7 could also cooperate with adenovirus type 5 E1B as well as with EJ-ras in transforming primary rat embryonal cells. However, E6/E7 could not cooperate with Ad12E1B in both cell systems. In this study, HPV 16 E6/E7 was assumed to have the growth-stimulatory activity in association with some cellular genes and transforming activity by cooperation with some transforming genes. These results suggest that E6/E7 seems to play a major role in the process of human cervical cell carcinogenesis. PMID- 2548944 TI - Malignant melanoma: a new mimic of colloid adenocarcinoma. PMID- 2548945 TI - Anaplastic large cell Ki-1 lymphoma. Delineation of two morphological types. AB - This report describes 16 cases of the recently recognized anaplastic large cell Ki-1 lymphoma. The disease showed a male:female ratio of 2.2:1 and a bimodal age distribution with peaks in the third and eighth decades. The clinical presentation was highly variable, with lymph node, skin, bone and gastrointestinal tract being the most commonly affected sites. The lymph nodes usually showed subtotal or sinusoidal involvement, and parenchymal fibrosis was common. The large neoplastic cells were almost invariably admixed with many reactive small lymphocytes, histiocytes and/or neutrophils. Two cytological types could be delineated: type I (pale cell, four cases) consisted of large polygonal cells with distinct pink-staining cell membrane and pale cytoplasm and pleomorphic nuclei showing marked chromatin clearing; and type II (basophilic cell, 12 cases) consisted of round or oval cells with basophilic cytoplasm and/or paranuclear pale hof, pleomorphic nuclei often reniform or lobulated and with frequent multinucleated wreath-like and Reed-Sternberg-like cells. Immunohistochemical studies showed that nine cases (56.3%) exhibited a T-cell phenotype, three cases (18.8%) each exhibited a B-cell or null-cell phenotype, while one case exhibited both T- and B-cell markers. Cutaneous involvement at presentation was associated with a favourable outcome, and spontaneous regression was common. For patients with non-cutaneous presentation, the prognosis was relatively good for young patients treated with aggressive chemotherapy, but was grave for old patients. PMID- 2548946 TI - Morphological and immunohistochemical investigation of non-Hodgkin's lymphoma combined with Hodgkin's disease. AB - Twenty cases with a morphological picture highly suspicious for a combination of non-Hodgkin's lymphoma and Hodgkin's disease were investigated. The infiltrates of Hodgkin's disease differed from those of non-Hodgkin's lymphoma in their cellular component of Hodgkin and Sternberg-Reed cells and the irregularity in the fibre pattern. Based upon histological and immunohistochemical criteria the 20 cases were divided into three groups. Group 1 (n = 10) contained seven chronic lymphocytic leukaemias of B type, one lymphoplasmacytoid immunocytoma, and two centroblastic/centrocytic lymphomas. The non-Hodgkin's lymphoma components showed a monotypic immunoglobulin distribution pattern and/or leukaemic blood picture. Adjacent to the non-Hodgkin's lymphoma was typical Hodgkin's disease in which Hodgkin and Sternberg-Reed cells were positive for both immunoglobulin light chains and IgG and reacted with anti-CD15. Group 2 (n = 5) consisted exclusively of centroblastic/centrocytic lymphoma in combination with Hodgkin's disease in which the few Hodgkin and Sternberg-Reed cells were negative with anti-CD15 monoclonal antibody. Group 3 (n = 5) consisted of four chronic lymphocytic leukaemias of B type and one lymphoplasmacytoid immunocytoma. In these cases no combination with Hodgkin's disease could be diagnosed apart from the presence of partially CD15 positive Hodgkin and Sternberg-Reed cells. The following conclusions were drawn: anti-CD15 (LeuM1 and 3C4/C3D-1) can neither confirm nor exclude Hodgkin's disease since, while they do not detect Hodgkin and Sternberg Reed cells in all cases of Hodgkin's disease, they do recognize Hodgkin and Sternberg-Reed cells in some B-cell lymphomas; anti-CD30 (Ber-H2) reacted with Hodgkin and Sternberg-Reed cells in all cases of Hodgkin's disease and also detected these cells in cases of non-Hodgkin's lymphoma. PMID- 2548947 TI - Two cases of pseudosarcomatous invasive transitional cell carcinoma of the urinary bladder mimicking malignant fibrous histiocytoma. PMID- 2548948 TI - Breakage of the human Y-chromosome short arm between two blocks of tandemly repeated DNA sequences. AB - Y-chromosomal rearrangements, a common cause of sex reversal in man, frequently occur between two blocks of repeated DNA. Both blocks are composed of 20-kb tandemly repeated Y-chromosome-specific DNA sequences. They are located in the proximal portion of the Y short arm on a NotI restriction fragment of approximately 5.3 Mb and on an MluI fragment of approximately 5.5 Mb. Chromosome breaks positioned between the two blocks were detected in two of three 46,XY females with deletions of Yp and in five of six 46,XX males positive for the repeat sequences. The rearranged NotI fragments in the 46,XX males were 4.4 Mb and the MluI fragments were 2.0 Mb in length. This indicates that breaks occur within a small region of Yp defined by the two blocks of specific repeated DNA sequences. The region between the two blocks thus appears to be a focus of structural lability in the human Y chromosome. PMID- 2548949 TI - A method for two-dimensional DNA electrophoresis (2D-DE): application to the immunoglobulin heavy chain variable region. AB - The scarcity of single-copy probes creates difficulty in the generation of large scale physical maps of mammalian gene families. A simple method of two dimensional DNA electrophoresis (2D-DE) has been developed to overcome this problem. DNA (2 micrograms) is digested with a rare-cutting restriction endonuclease and size separated by pulsed-field gel electrophoresis (PFGE). The DNA, still contained within the lane of the PFGE gel, is digested with a second frequent-cutting restriction enzyme and is subjected to an electrical field perpendicular to that of the PFGE. 2D-DE allows the simultaneous mapping, to large restriction fragments, of all the genes detected by a particular probe. The human immunoglobulin variable region was used as an example for this procedure. Two VH5 genes, on 8- and 9-kb EcoRI fragments, were mapped to 200- and 65-kb SfiI fragments, respectively, by 2D-DE. This technique will be particularly useful in the generation of physical maps of complex human gene families and of repeat families. PMID- 2548950 TI - The gene for the human putative apoE receptor is on chromosome 12 in the segment q13-14. AB - We have previously described the cDNA coding for a new lipoprotein receptor that contains domains closely related to the ligand-binding domain of the LDL receptor. We have now investigated the localization of the gene for this new receptor by hybridization of the cDNA to panels of rodent cells containing subsets of human chromosomes and by in situ hybridization of the cDNA to chromosomes. The gene maps to 12q13-14, a known hot spot for chromosomal rearrangements in human neoplasia. Of particular interest is the frequent involvement of the 12q13-14 segment in clonal abnormalities in lipomas and myxoid liposarcomas, and it is possible that LRP may play a role in the pathogenesis of such tumors. PMID- 2548951 TI - Abnormal rearrangements of T-cell receptor genes occur in long-term cultured bone marrow cells of lpr/lpr mice. AB - To search the abnormality in prethymic T-cell precursors in lpr/lpr(lpr) mice, rearrangement and expression of T-cell receptor (TcR) genes were investigated in long-term cultured bone marrow (LTBM) cells of lpr mice, in which the developmental steps of T-cell precursors may be better synchronized than those in bone marrow (BM) cells. Neither rearrangment nor expression of TCR gamma and delta genes were detected in the LTBM cells from +/+ control mice, whereas some gamma gene rearrangements were detected in those derived from lpr mice, irrespective of the genetic background. When BM cells or LTBM cells from lpr mice were transplanted into supralethally irradiated +/+ mice the lpr-derived BM cells appeared earlier in the thymus of the recipient mice than +/+-derived BM cells and the recipients suffered from lethal wasting syndrome. In addition, the lpr derived BM cells showed higher activity in colony-forming unit spleen (CFUs) than the +/+-derived BM cells. These results suggest that the T-cell progenitors in the BM of lpr mice may be different not only in quantity but also in quality from those of +/+ mice. PMID- 2548952 TI - Activation of macrophages to express cytocidal activity correlates with inhibition of their responsiveness to macrophage colony-stimulating factor (CSF 1): involvement of a pertussis toxin-sensitive reaction. AB - Bone marrow-derived murine macrophages were used to study the relationship between the proliferative response of macrophages to macrophage colony stimulating factor (CSF-1) and their activation for cytocidal activity against tumour cells. Macrophage activation required two sequential signals. Lymphokines (gamma interferon, interleukin-4) provided the first (priming) signal; bacterial products (lipopolysaccharide, lipophilic muramyl tripeptide, lipopeptide 31362, pertussis toxin) provided the second (triggering) signal. Both priming and triggering agents inhibited [3H]-thymidine uptake by macrophages stimulated with CSF-1. The antiproliferative activity of priming and triggering stimuli was synergistic. Pretreatment with triggering stimuli at 37 degrees C caused a rapid reduction of the subsequent binding of [125I]-CSF-1 to the cell surface at 4 degrees C, whereas priming agents had relatively little effect. Growth inhibition by both priming and triggering agents was largely reversible by washing the cells, and occurred even when they were added as long as 24 h after the growth factor. The ability of pertussis toxin to both inhibit CSF-1-induced proliferation and trigger cytotoxicity in macrophages suggests the involvement of a regulatory GTP-binding protein (G protein) in both processes. PMID- 2548953 TI - Comparative study on the antiviral activity of tumor necrosis factor (TNF)-alpha, lymphotoxin/TNF-beta, and IL-1 in WISH cells. AB - We find that pretreatment of WISH cells with tumor necrosis factor (TNF)-alpha, IL-1, and lymphotoxin/TNF-beta is capable of inducing an antiviral state in these cells, thereby protecting them from vesicular stomatitis virus cytopathic effect. Furthermore, we find that such a treatment causes a major inhibition of the synthesis of VSV proteins, as analyzed by SDS-PAGE. The 2-5A synthetase activity is also increased by treating the cells with doses of cytokines effective in antiviral protection. In this cell system, inclusion of polyclonal antibodies to IFN-beta during cytokine pretreatment abrogates the antiviral state elicited by the above cytokines, while antibodies to IFN-beta 2/IL-6 fail to abolish the cytokine-induced antiviral effects. PMID- 2548955 TI - Interleukin 6 primes human neutrophil and monocyte oxidative burst response. AB - Interleukin 6 (IL-6), a 26-kDa inducible protein, is a cytokine with multiple biological activities. This paper reports on the regulatory role of rIL-6 on the function of human polymorphonuclear and mononuclear leukocytes, a property not described previously. rIL-6 by itself did not exhibit any chemotactic activity and it could not activate these cells for an oxidative burst response. Preincubation of both cell types with rIL-6 at concentrations of 5 and 50 ng/ml primed the cells for enhanced generation of oxygen radicals following stimulation with the chemotactic peptide f-Met-Leu-Phe or the phorbol ester PMA. The enhancement of the oxidative burst response occurred both at the level of superoxide anion generation, an early step in the activation pathway, and at the level of the hydrogen peroxide-myeloperoxidase mediated response, a later step in the oxidative burst pathway. The priming ability was abolished by heat treatment of rIL-6 at 100 degrees C but not at 70 degrees C. Stimulation of B cell growth and immunoglobulin production combined with enhancement of oxidative burst response of phagocytic cells by IL-6 provide an effective mechanism of fighting against invading micro-organisms. PMID- 2548956 TI - Human peritoneal macrophage activity is increased by tuftsin. AB - Peritonitis caused by Candida albicans is a major complication of continuous ambulatory peritoneal dialysis (CAPD). Increasing the activity of the peritoneal macrophages--the predominant cell type found in the peritoneal cavity--may be of great importance in the prevention and therapy of peritonitis. Therefore, the activating effect of tuftsin was studied on human peritoneal macrophages from CAPD patients. Tuftsin induced a biphasic effect on macrophage activity within a range of 2 X 10(-9)-2 X 10(-6) M, with a maximal activity of 2 X 10(-7) M. At this concentration, tuftsin enhanced by twofold cell association with radiolabelled candida (from 2 +/- 0.2 to 4 +/- 0.2 candida per macrophage) and superoxide anion production in response to exposure to candida (from 150 +/- 20 to 300 +/- 20 nmoles/mg). These results suggest the potential use of tuftsin as a therapeutic drug. PMID- 2548954 TI - Characterization of the immune response to intracerebral inoculation of Theiler's murine encephalomyelitis virus: fine protein specificity of the splenic T cell and humoral antibody response. AB - The spleen T cell proliferative response of SJL/J mice primed by intracerebral inoculation of Theiler's murine encephalomyelitis virus (TMEV) was characterized in vitro. A dose-response [3H]thymidine incorporation was found with TMEV, but not with unrelated viruses. The maximum response was obtained at doses of 5 X 10(5) cells per well with cultures of 96 h duration. Viral capsid proteins VP1 and VP2 carried the antigenic determinants for the T cell response, VP3 playing no role in the in vitro cellular immune response. The antibody synthesis showed the same antigenic specificity as the cellular one, as only antibodies against VP1 and VP2 could be found in the sera of the immunized animals. PMID- 2548957 TI - [Leiomyoblastoma of the small intestine]. AB - A case of leiomyoblastoma in the small intestine is illustrated. The patient underwent first and iliac resection and then, 36 months later, a removal of multiple hepatic metastasis. The authors recall as the benign appearance of a leiomyoblastoma sometimes in only apparent, so that it's quite difficult predicting its evolution. Not any histological method can give express an available valuation about the neoplasm malignant potency. PMID- 2548958 TI - Multiplication of herpes simplex virus in large granular lymphocytes that co fractionate with human natural killer cell activity. AB - Highly enriched preparations of human large granular lymphocytes (LGL) cells, isolated from peripheral blood of normal adult donors, showed partial intrinsic resistance to infection with herpes simplex virus type 1 (HSV-1). Three subsets of LGL cells were identified on the basis of susceptibility to this virus: 1) resistant cells: 2) abortively infected cells; and 3) permissive cells. An average of 25% of LGL cells were completely resistant to infection. The majority (approximately 75%) could be infected as estimated by immunofluorescence. Only 5% of the original cell suspensions were productively infected as determined by infectious center assay and transmission electron microscopy. These results have been reproduced in multiple experiments from 8 different donors consisting of both males and females. No significant difference in LGL cell responses to HSV-1 were detected within this population. Enriched LGL preparations exhibited enhanced natural killer (NK) cell activity. These findings raise several questions concerning the biological significance of LGL susceptibility to infection with HSV-1, relative to virus transport and/or immune surveillance by NK cells. PMID- 2548959 TI - Central alpha-adrenoceptor influence over the cardiovascular reflexes activated by veratrine. AB - Intravenous veratrine induced alterations in cardiovascular parameters in cats were used as a tool for assessing the influence of central alpha-adrenoceptors over reflex adjustments in the heart rate and blood pressure. Blockade of central alpha 2-adrenoceptors with idazoxan or yohimbine, inhibited, while their activation by clonidine, as also blockade of alpha 1-adrenoceptors, with prazosin, potentiated the veratrine induced bradycardia. The hypotensive effect was relatively unaltered by these treatments. Low doses of clonidine potentiated the veratrine-induced bradycardia. It appears that alpha 2-adrenoceptor mechanisms exert greater control over the reflex regulation of heart rate than over reflex control of blood pressure. PMID- 2548961 TI - Role of angiotensin converting enzyme and other peptidases in in vivo metabolism of kinins. AB - Arterial plasma kinins and mean arterial pressure were measured in intact and bilaterally nephrectomized rats infused with vehicle or bradykinin to study the role of 1) angiotensin converting enzyme (ACE) and other peptidases and 2) the kidney (a kininase-rich organ) in the metabolism of kinins in vivo. Before the infusion, rats were pretreated with vehicle, enalaprilat (an ACE inhibitor), or a cocktail of kininase inhibitors containing 1) enalaprilat, 2) DL-2-mercaptomethyl 3-guanidinoethyl-thiopropanoic acid (MGTA), a carboxypeptidase N inhibitor, 3) phosphoramidon, a neutral endopeptidase 24.11 inhibitor, and 4) bestatin, an aminopeptidase B inhibitor. In the rats with vehicle (n = 8), the cocktail did not significantly increase endogenous kinins (from 31 +/- 6 to 41 +/- 9 pg/ml, p = 0.94). In the rats infused with bradykinin (peptidase substrate), plasma kinins increased threefold in the group pretreated with the vehicle, 21-fold in the enalaprilat group, and 22-fold in the cocktail group. These increases were doubled by nephrectomy but were not affected by ureteral ligation. In the groups pretreated with the cocktail or enalaprilat, the hypotensive effect of bradykinin was correlated with plasma kinin concentration (r = 0.75, p less than 0.001). After bradykinin infusion was stopped, plasma kinins decreased by half in 10-12 seconds in the rats pretreated with vehicle, enalaprilat, or cocktail. We concluded that ACE and the kidney are important to the metabolism of circulating kinins while carboxypeptidase N, neutral endopeptidase 24.11 and aminopeptidase B are not. We also concluded that other tissue peptidases, not affected by either the above inhibitors or nephrectomy, play an important role in kinin metabolism. PMID- 2548960 TI - Enhanced responsiveness to angiotensin II in vascular smooth muscle cells from spontaneously hypertensive rats is not associated with alterations in protein kinase C. AB - This study compares vascular smooth muscle cells from spontaneously hypertensive and normotensive Wistar-Kyoto rats with respect to protein kinase C and intracellular responses to angiotensin II (Ang II). Ang II-induced degradation of polyphosphoinositides and accumulation of inositol di- and tris-phosphates was enhanced (approximately twofold) in hypertensive-derived cells, without a change (vs. normotensive-derived cells) in half-maximally effective concentrations of Ang II. Intracellular pH (approximately 6.6) was comparable between both cell isolates at quiescence, but alkalinization induced by Ang II, serum, or phorbol ester was greater (delta 0.1-0.2 pH units) for hypertensive-derived cells. For both cell types, the intracellular pH response to these agonists was prevented in the presence of Na+-H+ exchange inhibitors. S6 kinase activation induced by Ang II was enhanced (approximately twofold) in hypertensive-derived cells, whereas activation in response to serum or 12-O-tetradecanoylphorbol 13-acetate did not differ significantly between the two cell types. Quantitation of protein kinase C by immunoblotting and [3H]phorbol dibutyrate binding procedures revealed no differences between the two smooth muscle cell isolates (at quiescence or in the presence of serum) with respect to either total amounts or subcellular distribution. Sensitivity of protein kinase C to phorbol ester was apparently also not different between the two cell types, as assessed from dose-dependent (phorbol ester) S6 kinase activation profiles. Phorbol ester caused a similar subcellular redistribution of [3H]phorbol dibutyrate binding in the two cell isolates, but for both, minimal (10%) translocation occurred in response to Ang II. The data suggest that enhanced agonist responsiveness in vascular smooth muscle cells is unlikely to involve alterations in protein kinase C. PMID- 2548962 TI - Astrocytes in cell culture incorporate GM1 ganglioside. AB - Ganglioside GM1 3H-labelled at the terminal galactose was added to astrocyte cell cultures. GM1 incorporation was studied in the two typical forms of astrocytes in cell culture of flat and stellate morphology. There was a strong time- and concentration-dependent increase in GM1 incorporation for both cell types of astrocytes. The incorporation of GM1 into the stellate form increased continuously up to 48 h (maximum time studied), while the incorporation into the flat form reached a plateau at the same time. After 2 h of GM1 incubation additional gangliosides appeared; the latter resulted from the metabolism of the GM1 incorporated, indicating that astrocytes in cell culture can biosynthesize more complex gangliosides. To confirm that GM1 was indeed incorporated into astrocytes, two other different approaches were used. Astrocyte cells treated with 3H-GM1 were visualized using autoradiography. The specific marker for GM1, rhodamine-labelled choleratoxin, was used to detect the incorporated GM1 using fluorescence microsocpy. In both cases GM1 treated cells were intensely labelled. These observations indicate that exogenous GM1 ganglioside can also be integrated into the astrocyte membranes as occurs in other types of cells and membranes. PMID- 2548963 TI - Combination therapy for systemic mycosis. PMID- 2548964 TI - Comparison of erythromycin ethylsuccinate and co-trimoxazole for treatment of pertussis. AB - Fifty-five ambulatory children with early culture-proven pertussis were treated for two weeks either with erythromycin ethylsuccinate (n = 28) (50-80 mg/kg/day in three doses during meals) or with co-trimoxazole (n = 27) (6-10 mg trimethoprim/kg/day in two doses after meals). After completion of treatment, all patients in the erythromycin group were culture-negative, while in the co trimoxazole group one child was still culture-positive. In this case vomiting may have played a role. Both agents appear to be able to eradicate Bordetella pertussis from the nasopharynx of patients with early whooping cough. PMID- 2548966 TI - Atrial natriuretic peptide: its mechanism of release from the atrium. PMID- 2548965 TI - Modulation of Clostridium perfringens intestinal colonization in infants delivered by caesarean section. AB - The colonization by Clostridium perfringens was investigated in 19 infants delivered by caesarean section during the two first weeks of life. The pattern of C. perfringens colonization depended upon the feeding. Breast feeding led to the repression of C. perfringens, whereas bottle feeding allowed its maintenance. On the contrary, Bifidobacterium bifidum growth was favoured by breast feeding. However, in one breast-fed infant, B. bifidum was never isolated and C. perfringens decreased. Breast feeding was able to directly modulate C. perfringens numbers. In fact, B. bifidum also had an effect, as demonstrated by the lower mean counts of C. perfringens, in bottle-fed infants carrying the bifidobacteria flora (p = 0.05). None of the bifidobacteria investigated in this study led to the same decrease. PMID- 2548967 TI - Double-blind comparison of itraconazole with griseofulvin in the treatment of tinea corporis and tinea cruris. AB - Seventy-eight patients with tinea corporis or tinea cruris participated in a double-blind study with either 100 mg itraconazole or 500 mg ultramicronized griseofulvin for 15 consecutive days. Clinical outcome was significantly in favor of itraconazole at completion of treatment (72% response rate vs. 51%) and at the follow-up visit (91% response vs. 64%). The most important difference between both treatments was the mycologic outcome, for which itraconazole showed a cure rate of 87% compared to 57% for griseofulvin 2 weeks after completion of therapy. It is suggested that 100 mg of itraconazole orally once daily is significantly more effective than 500 mg of griseofulvin once daily for 15 days in the treatment of glabrous skin infections. Both drugs were well tolerated. PMID- 2548968 TI - Small cell carcinoma of the ovary that is commonly associated with hypercalcemia is a neuroendocrine tumor on the basis of presently available evidence. PMID- 2548969 TI - [Leser-Trelat syndrome]. PMID- 2548970 TI - Elevated plasma cortisol levels during interferon-gamma treatment. AB - We investigated plasma levels of cortisol and ACTH in 9 patients with advanced metastatic tumors before and during treatment with interferon-gamma (IFN-gamma), 2-4 h after administration of IFN-gamma there was a sharp rise in plasma cortisol levels. The rise coincided with the onset of fever. Highest cortisol levels were reached 4-6 h after IFN-gamma injections, whereas IFN-gamma serum levels peaked 6 10 h after the IFN-gamma injections. Elevated cortisol levels during IFN-gamma treatment may be part of a homeostatic response of the neuroendocrine system to the immunological stimulus induced by IFN-gamma. On the other hand, the increased plasma levels of cortisol in response to pharmacological doses of IFN-gamma may dampen the in vivo effectiveness of IFN-gamma. PMID- 2548972 TI - Preparation of [11C]- and [125I]IMB: a dopamine D-2 receptor antagonist. AB - A new dopamine D-2 receptor ligand, N-[2-(N'-4-iodobenzyl-N'-methyl)aminoethyl]-5 chloro-2-methoxy-4- (methylamino)benzamide (IMB) was synthesized and labelled with 11C and 125I at high specific activities. Both radiotracers were prepared by alkylation reactions with [11C]iodomethane and [125I]4-iodobenzyl bromide and appropriate 2 degrees amine precursors. Isolated radiochemical yields of 12-20% for [11C]IMB and 35-50% for [125I]IMB were achieved in 24 and 130-150 min respectively. [11C]IMB and [123I]IMB may prove suitable for a direct comparison of PET and SPECT in the localization and quantification of neuroreceptors. PMID- 2548971 TI - Alloimmunization-induced changes in beta-adrenoceptor expression and cAMP on B lymphocytes. AB - In this report we have examined the effect of alloimmunization on beta-adrenergic expression in lymphocytes. We have observed a variation in the number of beta adrenoceptors (Bmax) according to the degree of immunization without modifications in their affinity (Kd). This phenomenon was accompanied with parallel variations of intracellular cAMP levels. A decrease in Bmax values was observed during the first and second immunizations. Then the Bmax began to increase, exceeding control values up to the fourth and fifth immunizations, and remaining constant at the sixth immunizations. Only B cell-enriched populations showed variation in Bmax values of beta-adrenoceptors with alloimmunization. In contrast, the Bmax values of T cell-enriched populations did not change. Kd values were similar in all cell types tested. The number of binding sites was not dependent on the animal's age. Modifications in cAMP levels of B cell-enriched populations were correlated with changes in beta-adrenoceptor expression. These results suggest that beta-adrenoceptor expression and cAMP intracellular levels in B cell-enriched populations vary with the number of alloimmunizations. In addition, the antibody synthesis induced by allogenic stimulus was inversely proportional to the number of beta-adrenoceptors expressed on B cells. Perhaps these findings are evidence of a control mechanism that regulates antibody synthesis during the immune response. PMID- 2548973 TI - Synthesis of 11C-labeled chlorpromazine directly from [11C]carbon dioxide. AB - Radiolabeled chlorpromazine was prepared by carboxylation of the N-trimethylsilyl derivative of norchlorpromazine with [11C]carbon dioxide, followed by in situ lithium aluminum hydride reduction. Radiochemical yields of 22-24% and radiochemical purities in the range of 93-98% were achieved. PMID- 2548975 TI - High specific activity 15-(p-[123I]iodophenyl)pentadecanoic acid. AB - An improved labeling procedure for 15-(p-[123I]iodophenyl)pentadecanoic acid (IPPA) is reported. The exchange labeling yield was greater than 95%. The radiochemical yield is influenced by the presence of sodium. The major advantage of this method is that very high specific activity i.e. upto 190 mCi/mg labeled IPPA can be prepared. PMID- 2548974 TI - Synthesis of the labeled D1 receptor antagonist SCH 23390 using [11C]carbon dioxide. AB - A new synthesis is described for the production of the positron emitting radiopharmaceutical R-(+)-7-chloro-8-hydroxy-2,3,4,5-tetrahydro-3-N-[11C]methyl-1 phenyl-1H- 3-benzazepine (SCH 23390, 2a). This novel method involves reductive carboxylation, in which [11C]CO2 is reacted with the trimethylsilyl derivative of the desmethyl compound (SCH 24518, 1a) followed by treatment with lithium aluminum hydride, to afford no carrier added 11C-labeled SCH 23390. The procedure gave chemically and radiochemically pure 11C-labeled SCH 23390 in 53-72% radiochemical yield with an unoptimized specific activity of 40 Ci/mmol within 45 50 min from the end of bombardment. PMID- 2548976 TI - A (re)investigation of the influence of some metal ions on the mean size of 99mTc(Sn)-MDP constituents at neutral pH. PMID- 2548977 TI - Computer-controlled radiochemical synthesis: a chemistry process control unit for the automated production of radiochemicals. AB - A computer-controlled general purpose chemistry process control unit (CPCU) suitable for the automated production of radiochemicals has been developed. This valve-and-tubing synthesis system can be user programmed to accommodate a variety of chemical processes. In a practical demonstration of its utility, the CPCU has been configured and programmed to synthesize 2-deoxy-2-[18F]fluoro-D-glucose (2 [18F]FDG) using aqueous [18F]fluoride ion. Using this instrument, the yield of 2 [18F]FDG from [18F]fluoride ion is 54.9% (+/- 11.2%, n = 125) corrected to EOB, after a synthesis time of 50-55 min. The average total activity produced (for runs of 5-10 microA) is 28.1 mCi/microA (+/- 5.03 mCi/microA). Thus, the amount of 2-[18F]FDG produced from a 10 microA for 1 h bombardment was 154.3 mCi (+/- 27.4 mCi). The unit has been similarly configured and programmed to synthesize 2 deoxy-2-[18F]fluoro-D-mannose (48% EOB), 3-(2'-[18F]fluoroethyl)spiperone (29% EOB), and [18F]fluoroacetate (66% EOB) from aqueous [18F]-fluoride ion, and 2 [18F]FDG from gaseous acetyl hypo[18F]fluorite (20% EOB). PMID- 2548979 TI - Methods of producing radionuclides for spectrometric gamma-ray sources and their standardization--2. Europium-152. AB - The radionuclide 152Eu was produced by irradiating 151Eu2O3 or 151EuCl3 enriched to 98.9% with thermal neutrons. 152Eu was separated from rare-earth impurities by extraction chromatography on a column using 0.6 M nitric acid eluent. The specific activity of the 152Eu was 1-10 Ci/g of Eu. The impurity of the 154Eu did not exceed 0.1%. The activity standardization of 152Eu was carried out by extrapolation of 4 pi (beta, e, x)-gamma coincidences. The relative gamma-ray intensities for 28 gamma-ray energies emitted in 152Eu decay have been measured. PMID- 2548978 TI - A diagnostic study of proton-beam irradiated water targets. AB - A non-destructive method was developed for assessing the average density of a small-volume water target during particle beam irradiation. Due to local boiling of the target water the particle beam penetrates through the target and further through the thin back wall of the target unit. The charged particle beam excites water molecules both in the target water and in the cooling water. A light guide was inserted into the target cooling water to transfer the light emitted during de-excitation to a photodiode which produced a voltage signal when the beam entered the cooling water. Beam current and target-water temperature were recorded simultaneously with the diode voltage signal. The light emitted in the target during irradiation was photographed through a glass window on the target. Photographic views of the target medium during irradiation are presented. Formation of voids in the beam path of the target was verified with the photographic technique. PMID- 2548980 TI - Evidence of conformational changes in the non-equivalent binding sites of human serum transferrin. AB - Samples of monoferric human serum transferrin have been prepared in which the iron occupies predominantly the N-site (sample A) and the C-site (sample B). 111In was then added in concentrations small enough to ensure that there was always an excess of specific binding sites. Because of the presence of apo transferrin in both the samples, the occupancy by 111In in the two sites was only 75-78% C-site in sample A and only 61-65% N-site in sample B. Time differential PAC spectra showed a transition in the quadrupole frequency which took place at different temperatures, approximately 275 K in sample A and between 290 and 305 K in sample B. Debye and Arrhenius plots of the temperature dependence of the correlation time associated with molecular reorientation indicated an effective molecular volume about 50% larger than that of the hydrated diferric molecule determined by "biochemical" methods, and an activation energy for re-orientation of approximately 0.065 eV. PMID- 2548981 TI - Measurement of natural radioactivity levels in Indian foodstuffs by gamma spectrometry. AB - Measurements of natural radioisotopes present in some of the foodstuffs which form the main components of the composite Indian diet are presented. Assessment of daily intake of these natural radionuclides has been made on the basis of the average daily intake of these food-items by the population of Bombay and its environment. The content of 40K, 226Ra and 228Th radioactivity varies from 45.9 to 649.0 Bq/kg, 0.01 to 1.16 Bq/kg and 0.02 to 1.26 Bq/kg, respectively. The average daily intakes of 40K, 226Ra and 228Th have been estimated as 105.6, 0.17 and 0.18 Bq d(-1), respectively for the period 1970-1982. PMID- 2548982 TI - Interactions between transposable elements for insertion in the Drosophila melanogaster genome. AB - Using in situ hybridization to polytene salivary gland chromosomes, we have registered the co-occurrences of insertions of the four mobile elements, copia, mdg-1, I and P in the whole genomes of 17 highly-inbred lines of Drosophila melanogaster (the insertions in the centromeric regions were excluded); these elements differ in structure, DNA sequence and profile of developmental transcription. The mdg-1 and P elements tend to avoid each other on the X chromosomes but not on the autosomes; copia and mdg-1, two copia-like elements, show an excess of co-occurrences on the 2L and 3R chromosome arms but not on the X chromosomes. The pairs mdg-1/I, I/copia, I/P and copia/P do not show any kind of interaction. Populational studies are thus necessary to obtain complete accurate information on interactions between transposable elements for their sites of insertion in a genome. PMID- 2548983 TI - Variability in gold bead density in cells. Quantitative immunocytochemistry. AB - Variability in gold bead distribution between individual cells was demonstrated in both pituitary melanotrophic cells immunocytochemically reacted for ACTH and in neurohypophysial terminals reacted for oxytocin-neurophysin. Gold beads were confined to the secretory granules compartment of both tissues. Density of gold beads in melanotrophic cells reacted for ACTH varied from 100-480 gold beads/microns 2. A much narrower range of gold beads distribution (460-900 gold beads/microns 2) was observed in axons of the neurohypophysis reacted with anti oxytocin-neurophysin. These results indicate that the labeling density varies from cell to cell (as well as axon terminals) within morphologically homogeneous populations. Thus, it may reflect certain physiological differences between cells. A suggestion is being made that mean gold bead density coefficient of variation should be calculated by comparison between individual cells. PMID- 2548985 TI - Nosocomial diarrhea associated with enterotoxigenic Clostridium perfringens infection in dogs. AB - A retrospective analysis of the medical records of 30 consecutive cases of diarrhea occurring in dogs that were hospitalized in a teaching hospital was performed. A prospective analysis of culture results for Clostridium perfringens of dogs with diarrhea were compared with those of a control nondiarrheal group. Hospital-acquired diarrhea in dogs was found to be associated with multiple serotypes of enterotoxigenic Clostridium perfringens. Other potential etiologic agents could not be isolated. Clinical signs were variable, and included mild depression, anorexia, and soft to watery diarrhea with or without frank blood, mucus, and tenesmus. Fever was not present. There were no hematologic or serum biochemical abnormalities, nor were there any consistent virologic or parasitologic findings. Salmonella spp or Campylobacter spp were not identified by fecal culture. No risk factors could be identified. A dog that was euthanatized on the day it developed diarrhea had intestinal histologic findings suggestive of clostridial enteritis. Dogs with diarrhea had significantly higher fecal clostridial counts than did dogs without diarrhea (mean log10 counts +/- SD = 6.34 +/- 1.79 vs 4.75 +/- 2.07). Enterotoxin was found in the feces of 41% of diarrheic dogs but in only 7% of dogs without diarrhea. PMID- 2548984 TI - Changes of glycoconjugates in human hepatocellular carcinoma. AB - Receptors of 12 lectins in 25 cases of human hepatocellular carcinomas (HCC) were histochemically investigated by avidin-biotin-peroxidase complex (ABC) method. Liver tissues of five cirrhotic patients and five normal subjects were used as controls. SJA receptor was absent both in HCC and controls, while LCA and PSA receptors were present in all tissues studied here. Receptors of DBA, PHA, PNA, UEAI and SBA which did not bind to normal, cirrhotic and pericarcinomatous liver tissues had the positive rates of 4%, 44%, 16%, 4% and 12% in HCC, respectively. Four lectins which strongly bound to the non-cancer liver tissues had their receptors in 96% (ConA, WGA, RCAI) and 36% (BSAI) of HCC. The pretreatment of tissue sections with neuraminidase abolished most of WGA receptors and exposed some PNA binding sites. There were many differences in lectin distribution between HCC and noncancer liver tissues. The changes of glycoconjugates in HCC were discussed. PMID- 2548986 TI - Food aversion learning: ability of lambs to distinguish safe from harmful foods. AB - We studied the ability of lambs to select safe foods in the presence of harmful foods. In a series of feeding experiments, 3- to 6-mo-old lambs were offered a choice between 1) a familiar, safe and a novel, harmful food, 2) a novel, safe and a novel, harmful food and 3) a familiar, harmful and a novel, safe food. All harmful foods were palatable feeds that had been treated with lithium chloride (LiCl), a non-lethal gastrointestinal poison. When lambs ingested a meal composed of a novel food containing LiCl and a familiar, safe food, lambs subsequently avoided the novel food. Lambs avoided the novel food even when the familiar food contained LiCl. The response of lambs varied when lambs were given a choice between a novel, safe food and a novel food containing LiCl. Lambs either avoided the novel food containing LiCl and ingested the safe food or they limited their intake of both novel foods. Their response was dependent on the novelty of the food containing LiCl. When a novel, palatable food contained 2% LiCl, lambs always ate some of the food, even after experiencing illness from ingesting it. Lambs experienced with foods containing LiCl displayed greater food neophobia than lambs naive to LiCl-treated foods. Thus, novelty was the major criterion that lambs used to associate foods with gastrointestinal illness. PMID- 2548987 TI - Chronic effects of recombinant porcine growth hormone on adrenal weight and activity in pigs. AB - This study was conducted to examine serum cortisol concentrations and adrenal cortisol output in pigs treated chronically with recombinant pGH (rpGH) at a maximally effective anabolic dose. Recombinant pGH (140 micrograms/kg body weight) was administered daily to eight barrows for 77 d. At slaughter, adrenal glands were removed, weighed and sliced; slices of fresh adrenal tissue were incubated for 1 h in the presence or absence of ACTH. Recombinant pGH increased adrenal weight by 39%. This change was accompanied by an inversely proportional reduction of in vitro cortisol output per gram of tissue, with the net result that total cortisol output per adrenal per kilogram of BW was unaltered, as was cortisol output in the presence of ACTH. Serum cortisol concentrations were measured in 10 barrows fitted with femoral artery catheters and treated daily with 0 or 140 micrograms rpGH/kg BW for 8 d. Basal and ACTH-stimulated cortisol concentrations were not altered by rpGH treatment. These results do not support our earlier speculation that a pGH-dependent increase in adrenal weight is associated with a chronic increase in adrenal activity, but rather demonstrate that corticosteroid output is tightly regulated and remains constant despite a marked increase in the size of the adrenal glands. PMID- 2548990 TI - Malignant round cell tumor of the thoracopulmonary region in an adolescent. PMID- 2548989 TI - Characterization of primary cell cultures derived from rat renal proximal tubules. AB - Proximal tubules were prepared from rat kidney cortex by collagenase digestion and purified by Percoll gradient centrifugation. Their enrichment was estimated by comparing the specific activities of various cell-specific enzymes in homogenates of renal cortex and of the isolated tubules. The tubules were cultured in a 50:50 mixture of Dulbecco's modified Eagle's and Ham's F12 media supplemented with insulin, transferrin, epidermal growth factor, hydrocortisone, and prostaglandin E1. After 2 to 3 d an extensive outgrowth of epithelial cells developed from the attached tubules. After 5 to 7 d near confluent monolayers were obtained. Hormonal responsiveness, marker enzyme activities, and transport properties were determined to further characterize the primary cultures. The cultured cells exhibited increased cyclic AMP production in response to parathyroid hormone but not calcitonin or vasopressin, consistent with the absence of cells derived from distal and collecting tubules. The cells also retained significant levels of 25-hydroxyvitamin D3-1 alpha-hydroxylase, alkaline phosphatase, and gamma-glytamyl-transpeptidase, three enzymes that are primarily associated with the proximal tubule. The cultured epithelial cells also exhibit a Na+-dependent phosphate and glucose transport systems. Therefore, the cells retain many functional properties that are characteristic of proximal tubules. Thus, the primary cultures should be suitable for the study of processes that occur specifically within this segment of the rat nephron. PMID- 2548988 TI - Effects of transforming growth factor beta on growth of human mammary epithelial cells in culture. AB - Normal human mammary epithelial cells (HMEC) from different individual reduction mammoplasty specimens were all growth inhibited, and showed a flattened, elongated morphology in response to human recombinant transforming growth factor beta 1 (TGF beta). The degree of growth inhibition varied among specimens, but none of the normal HMEC maintained growth in the continued presence of TGF beta. The degree of growth inhibition also varied with cell age in vitro, cells closer to senescence being more sensitive. TGF beta sensitivity was additionally assayed in two established cell lines derived from one of the reduction mammoplasty specimens after exposure to benzo(a)pyrene. Although varying degrees of growth inhibition and morphologic changes were observed in the cell lines, both lines contained populations that maintained active growth in the presence of TGF beta. Subclones of these lines demonstrated a great plasticity in their growth response to TGF beta, with individual clones ranging from strongly growth inhibited to nearly unaffected. These results suggest that multiple factors influence the extent of TGF beta-induced growth effects on both normal and transformed mammary epithelial cells, and that some of these factors may act through epigenetic mechanisms. PMID- 2548991 TI - Familial occurrence of paragangliomas. AB - The occurrence of paragangliomas in three members of the same family is reported. It concerned a left carotid body tumor in one patient, a left carotid body tumor in her older sister, and a concomitant jugulotympanic and vagal body tumor in their younger brother. The radiological diagnosis in these patients was based on the typical hypervascular character in specific locations both on angiography and computed tomography. The familial occurrence of these lesions should be kept in mind whenever a chemodectoma is found, even in the absence of clinical symptoms in the direct relatives of the patients. PMID- 2548992 TI - Adenylate cyclase and cyclic AMP phosphodiesterase in Bradyrhizobium japonicum bacteroids. AB - Adenylate cyclase and cyclic AMP (cAMP) phosphodiesterase have been identified and partially characterized in bacteroids of Bradyrhizobium japonicum 3I1b-143. Adenylate cyclase activity was found in the bacteroid membrane fraction, whereas cAMP phosphodiesterase activity was located in both the membrane and the cytosol. In contrast to other microorganisms, B. japonicum adenylate cyclase remained firmly bound to the membrane during treatment with detergents. Adenylate cyclase was activated four- to fivefold by 0.01% sodium dodecyl sulfate (SDS), whereas other detergents gave only slight activation. SDS had no effect on the membrane bound cAMP phosphodiesterase but strongly inhibited the soluble enzyme, indicating that the two enzymes are different. All three enzymes were characterized by their kinetic constants, pH optima, and divalent metal ion requirements. With increasing nodule age, adenylate cyclase activity increased, the membrane-bound cAMP phosphodiesterase decreased, and the soluble cAMP phosphodiesterase remained largely unchanged. These results suggest that cAMP plays a role in symbiosis. PMID- 2548993 TI - New method for generating deletions and gene replacements in Escherichia coli. AB - We describe a method for generating gene replacements and deletions in Escherichia coli. The technique is simple and rapid and can be applied to most genes, even those that are essential. What makes this method unique and particularly effective is the use of a temperature-sensitive pSC101 replicon to facilitate the gene replacement. The method proceeds by homologous recombination between a gene on the chromosome and homologous sequences carried on a plasmid temperature sensitive for DNA replication. Thus, after transformation of the plasmid into an appropriate host, it is possible to select for integration of the plasmid into the chromosome at 44 degrees C. Subsequent growth of these cointegrates at 30 degrees C leads to a second recombination event, resulting in their resolution. Depending on where the second recombination event takes place, the chromosome will either have undergone a gene replacement or retain the original copy of the gene. The procedure can also be used to effect the transfer of an allele from a plasmid to the chromosome or to rescue a chromosomal allele onto a plasmid. Since the resolved plasmid can be maintained by selection, this technique can be used to generate deletions of essential genes. PMID- 2548994 TI - Nucleotide sequence of the transcriptional control region of the osmotically regulated proU operon of Salmonella typhimurium and identification of the 5' endpoint of the proU mRNA. AB - Southern blot analysis of 15 proU transposon insertions in Salmonella typhimurium indicated that this operon is at least 3 kilobase pairs in length. The nucleotide sequence of 1.5-kilobase-pair fragment that contains the transcriptional control region of the proU operon and the coding sequences specifying 290 amino acids of the first structural gene of the operon was determined. The predicted amino acid sequence of the product of this gene shows extensive similarity to the HisP, MalK, and other proteins that are inner membrane-associated components of binding protein-dependent transport systems. S1 mapping and primer extension analysis of the proU mRNAs revealed several species with different 5' ends. Two of these endpoints are sufficiently close to sequences that have weak similarities to the consensus -35 and -10 promoter sequences that they are likely to define two transcription start sites. However, we cannot rule out the possibility that some or all of the 5' endpoints detected arose as a result of the degradation of a longer mRNA. The expression of proU-lacZ operon fusions located on plasmids was normal in S. typhimurium regardless of the plasmid copy number. The sequences mediating normal, osmoregulated expression of the proU operon were shown by subcloning to be contained on an 815-base-pair fragment. A 350-base-pair subclone of this fragment placed onto a lacZ expression vector directed a high-level constitutive expression of beta-galactosidase, suggesting that there is a site for negative regulation in the proU transcriptional control region which has been deleted in the construction of this plasmid. PMID- 2548996 TI - Isolation, nucleotide sequence, and preliminary characterization of the Escherichia coli K-12 hemA gene. AB - The Escherichia coli hemA gene, essential for the synthesis of 5-aminolevulinic acid (ALA), was isolated and sequenced. The following criteria identified the cloned gene as hemA. (i) The gene complemented a hemA mutation of E. coli. (ii) The gene was localized to approximately 26.7 min on the E. coli chromosomal linkage map, consistent with the location of the mapped hemA locus. Furthermore, DNA sequence analysis established that the cloned gene lay directly upstream of prfA, which encodes polypeptide chain release factor 1. (iii) Deletion of the gene resulted in a concomitant requirement for ALA. The hemA gene directed the synthesis of a 46-kilodalton polypeptide in maxicell experiments, as predicted by the coding sequence. The DNA and deduced amino acid sequences of the E. coli hemA gene displayed no detectable similarity to the ALA synthase sequences which have been characterized from a variety of organisms, but are very similar to the cloned Salmonella typhimurium hemA sequences (T. Elliott, personal communication). Results of S1 nuclease protection experiments showed that the hemA mRNA appeared to have two different 5' ends and that a nonoverlapping divergent transcript was present upstream of the putative distal hemA transcriptional start site. PMID- 2548995 TI - Positive regulation of glutamate biosynthesis in Bacillus subtilis. AB - Nitrogen source regulation of glutamate synthase activity in Bacillus subtilis occurs at the level of transcription of the gltA and gltB genes, which encode the two subunits of the enzyme. We show here that transcription of gltA requires the product of gltC, a gene whose transcription is divergent from that of gltA and whose transcriptional control sequences overlap those of gltA. gltC mutants had decreased, aberrantly regulated levels of glutamate synthase activity and decreased gltA mRNA. The gltC gene product could act in trans to complement both these defects. In addition, the gltC gene product repressed its own transcription. The DNA sequence of gltC revealed that its putative product is very similar to a number of positive regulatory proteins from gram-negative bacteria (the LysR family). PMID- 2548997 TI - Purification and properties of ferredoxin and rubredoxin from Butyribacterium methylotrophicum. AB - A ferredoxin and a rubredoxin from Butyribacterium methylotrophicum, which displays a carbonyl-dependent acetyl-coenzyme A synthesis, were purified to electrophoretic homogeneity. The two electron carriers showed absorption spectra similar to those in Clostridium species. The ferredoxin displayed absorption peaks at 280 and 391 nm, while rubredoxin displayed absorption peaks at 279, 382, and 482 nm. Minimum molecular weights calculated from the respective amino acid compositions were 5,727 for ferredoxin and 5,488 for rubredoxin, excluding iron and inorganic sulfur atoms. Both electron carriers were isolated as monomers, according to gel-filtration data. Electron spin resonance analysis revealed that the ferredoxin was a 2[4Fe-4S]-type and that both clusters had a midpoint redox potential value of -410 mV, whereas rubredoxin contained one acid-stable iron and had a redox value of -40 mV. The coupling of these electron carriers to hydrogenase and carbon monoxide dehydrogenase activities was investigated. Rubredoxin showed higher activity towards carbon monoxide dehydrogenase, whereas ferredoxin showed higher activity towards hydrogenase. PMID- 2548998 TI - Magnesium transport in Salmonella typhimurium: genetic characterization and cloning of three magnesium transport loci. AB - Salmonella typhimurium strains lacking the CorA Mg2+ transport system retain Mg2+ transport and the ability to grow in medium containing a low concentration of Mg2+. Mutagenesis of a corA strain followed by ampicillin selection allowed isolation of a strain that required Mg2+-supplemented media for growth. This strain contained mutations in at least two loci in addition to corA, designated mgtA and mgtB (for magnesium transport). Strains with mutations at all three loci (corA, mgtA, and mgtB) exhibited no detectable Mg2+ uptake and required 10 mM Mg2+ in the medium for growth at the wild-type rate. A wild-type allele at any one of the three loci was sufficient to restore both Mg2+ transport and growth on 50 microM Mg2+. P22 transduction was used to map the mgt loci. The mgtA mutation was located to approximately 98 map units (cotransducible with pyrB), and mgtB mapped at about 80.5 map units (near gltC). A chromosomal library from S. typhimurium was screened for clones that complemented the Mg2+ requirement of a corA mgtA mgtB mutant. The three classes of plasmids obtained could each independently restore Mg2+ transport to this strain and corresponded to the corA, mgtA, and mgtB loci. Whereas the corA locus of S. typhimurium is analogous to the corA locus previously described for Escherichia coli, neither of the mgt loci described in this report appears analogous to the single mgt locus described in E. coli. Our data in this and the accompanying papers (M. D. Snavely, J. B. Florer, C. G. Miller, and M. E. Maguire, J. Bacteriol. 171:4752-4760, 4761-4766, 1989) indicate that the corA, mgtA, and mgtB loci of S. typhimurium represent three distinct systems that transport Mg2+. PMID- 2548999 TI - Magnesium transport in Salmonella typhimurium: expression of cloned genes for three distinct Mg2+ transport systems. AB - In Salmonella typhimurium, the corA, mgtA, and mgtB loci are involved in active transport of Mg2+ (S. P. Hmiel, M. D. Snavely, C. G. Miller, and M. E. Maguire, J. Bacteriol. 168:1444-1450, 1988; S. P. Hmiel, M. D. Snavely, J. B. Florer, M. E. Maguire, and C. G. Miller, J. Bacteriol. 171:4742-4751, 1989). In this study, the gene products coded for by the corA, mgtA, and mgtB genes were identified by using plasmid expression in Escherichia coli maxicells. Complementation was assessed by introducing plasmids into a Mg2+-dependent corA mgtA mgtB strain and determining the ability of the plasmid to restore growth on medium without a Mg2+ supplement. Complementing plasmids containing corA expressed a 42-kilodalton (kDa) protein. This protein was not expressed by plasmids containing insertions or deletions that eliminated complementation. A plasmid containing mgtA expressed 37- and 91-kDa gene products. Data obtained with subclones and insertions in this plasmid indicated that plasmids expressing only the 91-kDa polypeptide complemented; plasmids that did not express this protein did not complement regardless of whether they expressed the 37-kDa protein. Plasmids carrying mgtB expressed a single protein of 102 kDa whose presence or absence correlated with the ability of the plasmid to complement the Mg2+-dependent triple mutant. Fractionation of labeled maxicells demonstrated that the 42-kDa corA, the 91-kDa mgtA, and the 102-kDa mgtB gene products are all tightly associated with the membrane, a location consistent with involvement in a transport process. These data provide further support the for existence of three distinct systems for Mg2+ transport in S. typhimurium. PMID- 2549000 TI - Genetic and molecular analyses of the gene encoding staphylococcal enterotoxin D. AB - The gene (entD) encoding staphylococcal enterotoxin D (SED) has been located on a 27.6-kilobase penicillinase plasmid designated pIB485. This plasmid was present in all SED-producing strains tested. The entD gene was cloned on a 2.0-kilobase DNA fragment and was expressed in Escherichia coli. Sequence analysis of this fragment revealed an open reading frame that encoded a 258-amino-acid protein that possessed a 30-amino-acid signal peptide. The 228-amino-acid mature polypeptide had a molecular weight of 26,360 and contained a high degree of sequence similarity to the other staphylococcal enterotoxins. S1 nuclease mapping showed that transcription of entD was initiated 266 nucleotides upstream from the translation start codon. The entD gene was also shown to be activated by the staphylococcal regulatory element known as agr. PMID- 2549001 TI - Method for selection of transposable DNA and characterization of a new insertion sequence, IS493, from Streptomyces lividans. AB - A method to select for transposable elements from Streptomyces spp. by using insertional inactivation of a repressor gene that functions in Escherichia coli was developed. Plasmid pCZA126, which can replicate in Streptomyces spp. or E. coli, contains a gene coding for the lambda cI857 repressor and a gene, under repressor control, coding for apramycin resistance. E. coli cells containing the plasmid are apramycin sensitive but become apramycin resistant if the cI857 repressor gene is disrupted. Plasmids propagated in Streptomyces spp. can be screened for transposable elements that have disrupted the cI857 gene by transforming E. coli cells to apramycin resistance. This method was used to isolate a new 1.6-kilobase insertion sequence, IS493, from Streptomyces lividans CT2. IS493 duplicated host DNA at the target site, had inverted repeats at its ends, and contained two tandem open reading frames on each strand. IS493 was present in three copies in the same genomic locations in several S. lividans strains. Two of the copies appeared to be present in regions of similar DNA context that extended at least 11.5 kilobases. Several other Streptomyces spp. did not appear to contain copies of IS493. PMID- 2549002 TI - Rhizobium leguminosarum exopolysaccharide mutants: biochemical and genetic analyses and symbiotic behavior on three hosts. AB - Ten independently generated mutants of Rhizobium leguminosarum biovar phaseoli CFN42 isolated after Tn5 mutagenesis formed nonmucoid colonies on all agar media tested and lacked detectable production of the normal acidic exopolysaccharide in liquid culture. The mutants were classified into three groups. Three mutants harbored Tn5 insertions on a 3.6-kilobase-pair EcoRI fragment and were complemented to have normal exopolysaccharide production by cosmids that shared an EcoRI fragment of this size from the CFN42 genome. The Tn5 inserts of five other mutants appeared to be located on a second, slightly smaller EcoRI fragment. Attempts to complement mutants of this second group with cloned DNA were unsuccessful. The mutations of the other two mutants were located in apparently adjacent EcoRI fragments carried on two cosmids that complemented those two mutants. The latter two mutants also lacked O-antigen-containing lipopolysaccharides and induced underdeveloped nodules that lacked nitrogenase activity on bean plants. The other eight mutants had normal lipopolysaccharides and wild-type symbiotic proficiencies on bean plants. Mutants in each of these groups were mated with R. leguminosarum strains that nodulated peas (R. leguminosarum biovar viciae) or clovers (R. leguminosarum biovar trifolii). Transfer of the Tn5 mutations resulted in exopolysaccharide-deficient R. leguminosarum biovar viciae or R. leguminosarum biovar trifolii transconjugants that were symbiotically deficient in all cases. These results support earlier suggestions that successful symbiosis with peas or clovers requires that rhizobia be capable of acidic exopolysaccharide production, whereas symbiosis with beans does not have this requirement. PMID- 2549003 TI - Transcription of pfl is regulated by anaerobiosis, catabolite repression, pyruvate, and oxrA: pfl::Mu dA operon fusions of Salmonella typhimurium. AB - Pyruvate formate-lyase (EC 2.3.1.54), a key enzyme in the anaerobic metabolism of Salmonella typhimurium, catalyzes the conversion of pyruvate to acetyl coenzyme A and formate. pfl::Mu dA operon fusions were isolated for the study of transcriptional regulation. pfl was transcribed both aerobically and anaerobically, but the activity increased about sixfold under anaerobic conditions. The addition of pyruvate, formate, and acetate in nutrient broth did not have any effect on the anaerobic expression of pfl. However, the addition of pyruvate to minimal glucose medium increased the anaerobic expression of pfl. The expression of pfl varied in different growth media. Anaerobic expression of pfl was lower when the culture was grown in minimal glucose medium than when it was grown in nutrient broth. When Casamino Acids (Difco Laboratories, Detroit, Mich.) were added to minimal glucose medium, the expression of pfl increased proportionally with the amount of Casamino Acids added. The transcription of pfl was positively controlled by the oxrA gene product and was affected by both the cya and crp mutations. However, mutations in genes affecting the cyclic AMP cyclic AMP receptor protein complex or oxrA could not completely abolish the anaerobic derepression of pfl. In merodiploid strains, pfl::Mu dA/F' pfl+, the beta-galactosidase activities were decreased. The mutations gyrA, oxrC, and oxrE, which affected anaerobic metabolism, did not affect anaerobic expression of pfl. PMID- 2549004 TI - Transformation of a conditionally peptidoglycan-deficient mutant of Staphylococcus aureus with plasmid DNA. AB - A simple and reliable method for polyethylene glycol-induced plasmid transformation of a temperature-sensitive peptidoglycan-deficient mutant of Staphylococcus aureus is described. The procedure uses strains carrying the tofA372 mutation grown under conditions that yield osmotically fragile cells capable of efficient wall regeneration. The peptidoglycan-deficient cells were transformed with plasmids pE194 and pI258 at frequencies comparable with those obtained with protoplasts prepared with lysostaphin treatment. A readily portable tofA372 mutation was constructed by isolating an insertion of the erythromycin resistance transposon Tn551 adjacent to tofA372. tofA372 was shown by protoplast fusion and transformation analyses to be in the gene order hly-421-omega [Chr::Tn551]1059-tofA372-uraB232-omega [Chr::Tn916]1101-thrB106 on the chromosome of S. aureus NCTC 8325. PMID- 2549005 TI - Genes downstream from pucB and pucA are essential for formation of the B800-850 complex of Rhodobacter capsulatus. AB - The formation of the light-harvesting complex B800-850 (LH-II) of Rhodobacter capsulatus requires, in addition to the synthesis of the polypeptides alpha and beta (the gene products of pucA and pucB), the synthesis of bacteriochlorophyll and carotenoids and the expression of at least one gene localized downstream from the pucBA operon. This was concluded from the observation that a Tn5 insertion downstream from pucBA inhibited the formation of the LH-II complex and the formation of the pucBA mRNA. The Tn5 insertion point was mapped and found to be over 500 base pairs (bp) downstream from the end of the pucA gene, suggesting the presence of additional puc genes. A region of about 3,000 bp including the pucB and pucA genes and DNA downstream from pucA was sequenced and found to contain three open reading frames (ORFs C, D, and E). The polypeptide deduced from the first ORF (C) contains 403 amino acids with strongly hydrophobic stretches and one large and three small hydrophilic domains carrying many charged residues. The other two ORFs contain 113 (D) and 118 (E) codons. The amino acid sequences of the N terminus and two tryptic peptides of an alkaline-soluble Mr-14,000 subunit of the isolated LH-II complex were identical with the deduced amino acid sequence of ORF E. PMID- 2549007 TI - Structure and expression of the cytochrome aa3 regulatory gene ctaA of Bacillus subtilis. AB - Mutations that define the ctaA gene of Bacillus subtilis block cytochrome aa3 formation and sporulation. We have recently described the isolation and initial characterization of the ctaA locus. Analysis of in vivo mRNA transcripts by RNase protection experiments located the 5' and 3' termini of the ctaA transcript, confirming a monocistronic structure. By using a nuclease protection assay, an increase in the abundance of steady-state ctaA mRNA was observed during the initiation of sporulation, followed by a decrease during subsequent stages. Transcripts originating from the ctaA gene were most abundant 2.0 h after the end of exponential growth. This pattern of ctaA mRNA accumulation was confirmed by coupling the transcription of the ctaA gene to lacZ in an integrative plasmid vector. Expression of ctaA was not repressed by glucose and was independent of the spoOA and spoOH (sigH) gene products. Postexponential expression was found to be dependent on the product of the strC gene. The expression of ctaA appears to be regulated in a growth stage-specific manner. The transcriptional start site, identified by high-resolution S1 nuclease protection experiments, was preceded by a single sigma A-dependent promoter sequence. PMID- 2549006 TI - Isolation and sequence of ctaA, a gene required for cytochrome aa3 biosynthesis and sporulation in Bacillus subtilis. AB - Cytochrome aa3 is one of two terminal oxidase complexes in the Bacillus subtilis electron transport chain. A novel genetic strategy was devised which permitted the isolation of B. subtilis mutants lacking cytochrome aa3 by selection for streptomycin-resistant clones which failed to oxidize the artificial electron donor N,N,N',N'-tetramethyl-p-phenylenediamine. Two mutations were studied intensively. Spectroscopic examination showed that each mutant lacked cytochrome aa3; they were also asporogenous and unable to grow on lactate as the sole carbon and energy source. These mutations were mapped to a locus designated ctaA, located at 127 degrees between pyrD and metC on the B. subtilis chromosome. Both ctaA mutations were closely linked by transformation to the pycA locus. The ctaA locus and a portion of the pycA locus were cloned from a B. subtilis integration library constructed in Escherichia coli. A recombinant plasmid containing a 4.0 kilobase insert of B. subtilis DNA could transform both ctaA mutants to CtaA+. Gene disruption and complementation experiments with subcloned fragments revealed that the ctaA locus consisted of a single transcriptional unit about 1.35 kilobase pairs in size. The nucleotide sequence of the ctaA transcriptional unit contains a single open reading frame capable of coding for a protein with a predicted molecular weight of 34,065. The predicted protein is extremely hydrophobic, with several probable membrane-spanning domains. No sequence similiarity was found between ctaA and the highly conserved procaryotic and mitochondrial oxidase polypeptides. Cloning and sequence analysis of two ctaA mutations revealed that one allele is a nonsense mutation in the carboxy terminus and the other is a missense mutation in the amino terminus; this indicates that the pleiotropic phenotype conferred by each mutation was caused by loss of CtaA or of its activity. Genetic evidence suggests that the ctaA gene product is required as an accessory protein in the genetic expression, posttranslational biogenesis, or both, of the cytochrome aa3 complex and during an early stage of sporogenesis. PMID- 2549008 TI - Variation in buoyant density of whole cells and isolated cell walls of Streptococcus faecium (ATCC 9790). AB - The buoyant density of whole cells of Streptococcus faecium varies with growth rate and during the cell cycle. Two possible explanations for this were explored: (i) the density of cell walls may vary, and (ii) the proportions of wall and cytoplasm may vary. We tested the first possibility by isolating walls from chilled, unfixed populations of S. faecium cells and fractionating them on Percoll density gradients. Mean cell wall density averaged 4% less than whole cell density and did not vary significantly with growth rate. In addition, walls isolated from heavy and light fractions of a population of cells did not differ significantly in density. Thus, variation in the density of isolated cell walls could not account for the observed variation in whole-cell density within or between populations. Using previously published measurements of the physical dimensions of S. faecium cells, we also found that the relative proportions of wall and cytoplasm (see the second possibility above) could not account for the observed changes in whole-cell buoyant density. PMID- 2549009 TI - Analysis of the transcriptional unit encoding the genes for rubredoxin (rub) and a putative rubredoxin oxidoreductase (rbo) in Desulfovibrio vulgaris Hildenborough. AB - The nucleotide sequence of a 2.0-kilobase-pair EcoRI restriction fragment upstream from the gene (rub, 162 base pairs) encoding rubredoxin from Desulfovibrio vulgaris Hildenborough indicates that it is part of a larger transcriptional unit, containing an additional 378-base-pair open reading frame which terminates 16 nucleotides from the translational start of the rub gene and could encode a polypeptide of 14 kilodaltons (kDa). Northern (RNA) blotting of RNA isolated from both D. vulgaris Hildenborough and Escherichia coli TG2 transformed with plasmid pJK29, which contains both genes on a 1.1-kilobase-pair SalI insert, confirms that the genes for this 14-kDa polypeptide and rubredoxin are present on a single transcript of 680 nucleotides. Strong evidence that the 14-kDa polypeptide is also a redox protein is provided by the fact that its NH2 terminus is homologous to desulforedoxin, which has been isolated from D. gigas as a small dimeric redox protein (36 amino acids per monomer), coordinating two iron atoms. Since rubredoxin is a potential redox partner for the 14-kDa protein, it has been tentatively named rubredoxin oxidoreductase, produced by the rbo gene. Southern blotting indicates that the rbo-rub operon is present in several species and strains of sulfate-reducing bacteria. PMID- 2549010 TI - Characterization of five genes in the upper-pathway operon of TOL plasmid pWW0 from Pseudomonas putida and identification of the gene products. AB - The upper operon of the TOL plasmid pWW0 of Pseudomonas putida encodes a set of enzymes which transform toluene and xylenes to benzoate and toluates. The genetic organization of the operon was characterized by cloning of the upper operon genes into an expression vector and identification of their products in Escherichia coli maxicells. This analysis showed that the upper operon contains at least five genes in the order of xylC-xylM-xylA-xylB-xylN. Between the promoter of the operon and xylC, there is a 1.7-kilobase-long space of DNA in which no gene function was identified. In contrast, most of the DNA between xylC and xylN consists of coding sequences. The xylC gene encodes the 57-kilodalton benzaldehyde dehydrogenase. The xylM and xylA genes encode 35- and 40-kilodalton polypeptides, respectively, which were shown by genetic complementation tests to be subunits of xylene oxygenase. The structural gene for benzyl alcohol dehydrogenase, xylB, encodes a 40-kilodalton polypeptide. The last gene of this operon is xylN, which synthesizes a 52-kilodalton polypeptide of unknown function. PMID- 2549011 TI - Cloning and expression of plasmid genes encoding resistances to chromate and cobalt in Alcaligenes eutrophus. AB - Resistances to chromate and cobalt were cloned on a 30-kilobase-pair (kb) DNA region from the large Alcaligenes eutrophus plasmid pMOL28 into the broad-host range mobilizable cosmid vector pVK102. A restriction nuclease map of the 30-kb region was generated. The resistances expressed from the hybrid plasmids after transfer back into A. eutrophus were inducible and conferred the same degree of resistance as the parent plasmid pMOL28. Resistances were expressed in metal sensitive Alcaligenes strains and related bacteria but not in Escherichia coli. Resistance to chromate was further localized on a 2.6-kb EcoRI fragment, and resistance to cobalt was localized on an adjoining 8.5-kb PstI-EcoRI fragment. When the 2.6-kb EcoRI fragment was expressed in E. coli under the control of a bacteriophage T7 promoter, three polypeptides with molecular masses of 31,500, 21,000, and 14,500 daltons were visible on autoradiograms. The 31,500- and 21,000 dalton polypeptides were membrane bound; the 14,500-dalton polypeptide was soluble. PMID- 2549012 TI - Cloning of pMOL28-encoded nickel resistance genes and expression of the genes in Alcaligenes eutrophus and Pseudomonas spp. AB - The 163-kilobase-pair (kb) plasmid pMOL28, which determines inducible resistance to nickel, cobalt, chromate, and mercury salts in its native host Alcaligenes eutrophus CH34, was transferred to a derivative of A. eutrophus H16 and subjected to cloning procedures. After Tn5 transposon mutagenesis, restriction endonuclease analysis, and DNA-DNA hybridization, two DNA fragments, a 9.5-kb KpnI fragment and a 13.5-kb HindIII fragment (HKI), were isolated. HKI contained EK1, the KpnI fragment, as a subfragment flanked on both sides by short regions. Both fragments were ligated into the suicide vector pSUP202, the broad-host-range vector pVK101, and pUC19. Both fragments restored a nickel-sensitive Tn5 mutant to full nickel and cobalt resistance. The hybrid plasmid pVK101::HKI expressed full nickel resistance in all nickel-sensitive derivatives, either pMOL28-deficient or defective, of the native host CH34. The hybrid plasmid pVK101::HKI also conferred nickel and cobalt resistance to A. eutrophus strains H16 and JMP222, Alcaligenes hydrogenophilus, Pseudomonas putida, and Pseudomonas oleovorans, but to a lower level of resistance. In all transconjugants the metal resistances coded by pVK101::HKI were expressed constitutively rather than inducibly. The hybrid plasmid metal resistance was not expressed in Escherichia coli. DNA sequences responsible for nickel resistance in newly isolated strains showed homology to the cloned pMOL28-encoded nickel and cobalt resistance determinant. PMID- 2549013 TI - Legionella micdadei protein kinase catalyzes phosphorylation of tubulin and phosphatidylinositol. AB - Legionella micdadei, a pathogen which enters into host phagocyte phagolysosomal structures, contains at least two protein kinases. We have purified to homogeneity the predominant, nucleotide-independent protein kinase and examined its ability to catalyze the transfer of phosphate from ATP to acceptors in human neutrophils. The L. micdadei protein kinase catalyzed the phosphorylation of proteins of 11.5, 14, 19, 23, 28, 34, and 38 kilodaltons (kDa) present in a Triton X-100 extract of neutrophil membranes and of 11.5, 13.5, 25, and 38 kDa in the neutrophil cytosol. Tubulin was a good substrate for the L. micdadei protein kinase in vitro. The bacterial kinase also catalyzed the phosphorylation of phosphatidylinositol (PI) at about half the rate at which histones were phosphorylated; phosphatidylinositol-4-phosphate (PIP) was not phosphorylated by the kinase. The PI kinase activity of the L. micdadei enzyme was optimum at pH 7.0, and the divalent cation requirement was satisfied best by Mg2+ and Ca2+. The maximum rate of PI phosphorylation was obtained with 0.6 mM PI; in the presence of MgCl2 (10 mM), the Km for PI was 0.9 mM and the Km for ATP was 1.5 mM. The detergents octyl-beta-D-glucoside (10 to 20 mM) and Triton X-100 (0.5%) stimulated kinase activity twofold when PI was the phosphate acceptor; however, only octyl glucoside stimulated histone kinase activity. Various membrane phospholipids inhibited PI kinase activity. The most potent phospholipid inhibitor was the product of the PI kinase reaction, PIP, which at a 0.6 mM concentration inhibited both PI and tubulin phosphorylation by 80%. The inhibition of kinase activity by PIP when histone served as the acceptor was noncompetitive in character. The L. micdadei kinase also phosphorylated PI in intact. (3H)inositol-labeled neutrophils. The PI kinase and histone kinase activities of teh L. micdadei kinase copurified and cofucused (pI, 5.8) when subjected to isoelectric focusing, suggesting that the two enzymatic activities reside in a single protein. PMID- 2549014 TI - Insertion elements and deletion formation in a halophilic archaebacterium. AB - Deletion events that occur spontaneously in 36-kilobase-pair (kbp) plasmid pHH4 from the archaebacterium Halobacterium halobium were investigated. Four different deletion derivatives with sizes ranging from 5.7 to 17 kbp were isolated. Three of these deletion variants derived from pHH4 (pHH6 [17 kbp], pHH7 [16 kbp], and pHH8 [6.3 kbp]), whereas the 5.7-kbp plasmid pHH9 derived from pHH6. Strains containing pHH6, pHH7, or pHH9 each lacked the parental plasmid pHH4, while pHH8 occurred at a 1:1 ratio together with pHH4. Common to all of these plasmids was the 5.7-kbp region of pHH9 DNA. The regions containing the fusion site in the deletion derivatives were investigated and compared with the corresponding area of the parental plasmid. Each deletion occurred exactly at the terminus of an insertion element. In pHH6 and pHH7, a halobacterial insertion element (ISH2) was located at the deletion site. The DNA fused to ISH2 displayed a 7-base-pair (bp) (pHH7) or 10-bp (pHH6) sequence homology to the inverted repeat of ISH2. In the two smaller plasmids, pHH8 and pHH9, an ISH27 element was located at the deletion site. Most likely, all of these smaller plasmids resulted from an intramolecular transposition event. The ISH27 insertion sequence contains a 16-bp terminal inverted repeat and duplicates 5 bp of target DNA during the transposition with the specificity 5'ANNNT3'. Four ISH27 copies were analyzed, and two ISH27 element types were identified that have approximately 85% sequence similarity. The ISH27 insertion elements constitute a family which is related to the ISH51 family characterized for H. volcanii, another halophilic archaebacterium. PMID- 2549015 TI - Glycogen-bound polyphosphate kinase from the archaebacterium Sulfolobus acidocaldarius. AB - Glycogen-bound polyphosphate kinase has been isolated from a crude extract of Sulfolobus acidocaldarius by isopycnic centrifugation in CsCl. Divalent cations (Mn2+ greater than Mg2+) stimulated the reaction. The enzyme does not require the presence of histones for its activity; it is inhibited strongly by phosphate and slightly by fluoride. The protein from the glycogen complex migrated in a sodium dodecyl sulfate-polyacrylamide gel as a 57-kilodalton protein band; after isoelectric focusing it separated into several spots in the pH range of 5.6 to 6.7. PMID- 2549016 TI - Levels of DNA topoisomerases, single-stranded-DNA-binding protein, and DNA polymerase I in rho+ and rho-15 strains of Escherichia coli. AB - The Escherichia coli rho-15 mutant, which is highly defective in transcription termination, was examined to see whether its reduced DNA superhelicity could be explained by altered expression of proteins that may affect DNA structure. Levels of DNA gyrase and topoisomerase I were normal; levels of single-stranded-DNA binding protein, DNA polymerase I, and a protein tentatively identified as Lon were significantly altered. PMID- 2549018 TI - Fluoxetine-induced akathisia: clinical and theoretical implications. AB - Five patients receiving fluoxetine for the treatment of obsessive compulsive disorder or major depression developed akathisia. The typical fluoxetine-induced symptoms of restlessness, constant pacing, purposeless movements of the feet and legs, and marked anxiety were indistinguishable from those of neuroleptic-induced akathisia. Three patients who had experienced neuroleptic-induced akathisia in the past reported that the symptoms of fluoxetine-induced akathisia were identical, although somewhat milder. Akathisia appeared to be a common side effect of fluoxetine and generally responded well to treatment with the beta adrenergic antagonist propranolol, dose reduction, or both. The authors suggest that fluoxetine-induced akathisia may be caused by serotonergically mediated inhibition of dopaminergic neurotransmission and that the pathophysiology of fluoxetine-induced akathisia and tricyclic antidepressant-induced "jitteriness" may be identical. PMID- 2549017 TI - Orientation of IS50 transposase gene and IS50 transposition. AB - Reversal of transposase gene orientation with respect to the nonidentical ends of IS50 strongly decreased IS50 transposition in both Dam- and Dam+ hosts. In either orientation, IS50 transposase expression was unaffected. These effects were independent of the surrounding DNA context. This shows that the efficiency of IS50 transposition is dependent on transposase gene orientation. The transposition frequencies of transposons utilizing inverted IS50 inside ends (IE), IE-IE transposons, were lower than either outside end (OE)-IE or OE-OE transposons. PMID- 2549019 TI - The immunopotentiating property of lipophilic muramyl dipeptide and its molecular state in liposomal membranes: plaque-forming cell responses and ESR studies. AB - The immunopotentiating property of spin-labeled lipophilic muramyl dipeptide (SL MDP) in liposomes was studied as to the plaque-forming cell (PFC) response to glycophorin, as an antigen, in membranes. The effect of SL-MDP depended on the densities of both SL-MDP itself and the antigen. The addition of the optimal amount of SL-MDP to liposomes containing a low density (0.016 mol%) of the antigen increased the PFC response by three times, whereas the presence of SL-MDP in optimal antigen density (0.032 0.127 mol%) membranes was rather inhibitory. In these liposomes, the amounts and molecular states of SL-MDP were determined from ESR spectra and are discussed in connection with its immunopotentiating property. PMID- 2549021 TI - Localization of the NADH kinase in the inner membrane of yeast mitochondria. AB - The bulk of NADH kinase of Saccharomyces cerevisiae was recovered in the mitochondrial fraction prepared from spheroplasts. Most of the NADH kinase was localized in the inner membrane fraction, which was separated from other mitochondrial components by the combined swelling, shrinking, and sonication procedure. Treatment of mitoplasts with antiserum against the NADH kinase caused inactivation of the enzyme. On the contrary, no influence was observed upon the same treatment of intact mitochondria. p-Chloromercuribenzoate and eosin-5 maleimide inactivated the enzyme without affecting the matrix ATPase. The NADH kinase was enzymatically iodinated in mitoplasts, but not in the intact mitochondria. These results support the conclusion that NADH kinase is localized and functions at the intermembrane space side of the mitochondrial inner membrane. It is evident that the NADH kinase is encoded by nuclear gene(s) because it is synthesized in the presence of chloramphenicol or acriflavine, and a significant amount of the enzyme was detected in mitochondrial DNA-deficient mutants. PMID- 2549020 TI - Reevaluation of the spin-trapped adduct formed from 5,5-dimethyl-1-pyrroline-1 oxide during the respiratory burst in neutrophils. AB - By employing EPR spectrometry with the aid of a spin-trapping agent, 5,5-dimethyl 1-pyrroline-1-oxide (DMPO), the generation of superoxide anion and hydroxyl radical was reevaluated during the respiratory burst of porcine and human neutrophils. Properly prepared resting neutrophils did not generate any spin trapped radical, and, when the cells were stimulated with phorbol myristate acetate, only DMPO-OOH, the spin-trapped adduct of superoxide anion, was detected. No formation of DMPO-OH, the spin-trapped adduct of the hydroxyl radical, was observed. DMPO-OOH was also detected principally when the neutrophils were stimulated with opsonized zymosan, a particulate stimulus. In the latter case, however, the formation of DMPO-OOH ceased shortly after the addition of zymosan and subsequent production of DMPO-OH was observed. The production of DMPO-OH was found to be associated with cell injury. DMPO at the concentration usually used for the experiment (0.045-0.09 M) injured phagocytizing neutrophils, causing lysis of the cells. On the other hand, an addition of cell homogenate or glutathione-glutathione peroxidase system to the suspension of intact cells which were producing DMPO-OOH resulted in the formation of DMPO-OH. Thus, DMPO-OH was probably derived from DMPO-OOH by the action of enzymes and/or factor(s) which were released from the lysed cells. PMID- 2549023 TI - Enhanced bacteriolytic activity of hen egg-white lysozyme due to conversion of Trp62 to other aromatic amino acid residues. AB - Tryptophan at the 62nd position (Trp62) of hen egg-white lysozyme is an amino acid residue whose action is essential for its enzymatic activity. Its indole ring may possibly come into direct contact with sugar residues of the substrate, and thus contribute significantly to substrate binding. For further elucidation of its role in catalytic processes, this amino acid was converted to other aromatic residues, such as Tyr, Phe, and His, by site-directed mutagenesis. All the mutations were found to enhance the bacteriolytic activity but to decrease the hydrolytic activity toward an artificial substrate, glycol chitin. Such a change in substrate preference appears remarkable considering the smaller size of the aromatic residue on the mutant enzyme at the 62nd position. PMID- 2549024 TI - Regional odontodysplasia: new histopathological data. AB - Light microscopy, microradiography, SEM and TEM of 4 tooth follicles in a 12-year old caucasian girl presenting regional odontodysplasia showed widespread globular dentin, calcifications located in the enlarged pulp chambers, hypoplastic and hypomineralized enamel. Hypomineralized strands were sandwiched between two normal enamel layers, which indicates that amelogenesis, interrupted for a while, has once more become established. The enamel surface was covered with calcoglobules. Numerous rounded calcifications were scattered within the dental follicles. Some of these occurred in microfibrils (possibly oxytalan fibers), distinct from collagen fibers. Calcification of the sheath surrounding epithelial rests was a conspicuous feature. The fibroblasts in contact with calcifications developed numerous cytoplasmic extensions, which suggests that they may have assumed a phagocytosis behaviour. PMID- 2549022 TI - Orientation and reactivity of NADH kinase in proteoliposomes. AB - NADH kinase was reconstituted in liposomes by employing phosphatidylcholine and phosphatidylethanolamine with n-octyl-beta-D-thioglucoside as a detergent. An analogous molecular organization of the NADH kinase to that in the mitochondrial inner membrane was ascertained to exist in the liposomal membrane. Michaelis constants for NADH and ATP were determined as 27 and 133 microM, respectively. Both values were lower than that of the solubilized enzyme. The catalytic center of NADH kinase was exposed on the outer surface of the reconstituted liposomes. The NADH kinase reconstituted with ADP/ATP carrier protein catalyzed the phosphorylation of exogenously supplied NADH by the use of ATP entrapped in the liposomal matrix. PMID- 2549025 TI - Ubiquitin is involved in the in vitro insertion of monoamine oxidase B into mitochondrial outer membranes. AB - Monoamine oxidase B that has been synthesized by a reticulocyte lysate charged with bovine liver RNA will insert in a proteinase K-resistant form into isolated outer membranes from rat liver mitochondria. It appears that ubiquitin, a 76 amino acid polypeptide which is enzymatically conjugated to proteins, may be involved in the insertion process. Depletion of endogenous ubiquitin from the reticulocyte lysate with purified antibodies against this polypeptide inhibits the insertion of monoamine oxidase B, and this inhibition is relieved if ubiquitin is restored. On the other hand, a mutant form of ubiquitin which is unable to conjugate with proteins will not support insertion. Conjugation with ubiquitin is an ATP-dependent process. Not only does enzymatic depletion of ATP from the lysate prevent the insertion of monoamine oxidase, but ubiquitin will not restore insertion unless ATP is also present. These data indicate that the formation of a ubiquitin conjugate is involved in the insertion of newly synthesized monoamine oxidase B into the outer membranes. PMID- 2549026 TI - Ligand inhibition of the platelet glycoprotein IIb-IIIa complex function as a calcium channel in liposomes. AB - Platelet glycoproteins IIb and IIIa function as a fibrinogen receptor on the activated platelet. We have shown that these glycoproteins can be incorporated onto the surface of phosphatidylcholine vesicles with retention of fibrinogen and antibody binding properties and can permit Ca2+ transit across the phospholipid bilayer. In the current study we demonstrate that this apparent Ca2+ channel function is specifically inhibited by the synthetic analogue of the fibrinogen gamma COOH-terminal peptide, His-His-Leu-Gly-Gly-Ala-Lys-Gln-Ala-Gly-Asp-Val (His 12-Val), but not by the adhesive protein sequence Arg-Gly-Asp-Ser (RGDS). Prior incubation of IIb-IIIa liposomes with RGDS prevented Ca2+ transit inhibition by 25 microM His-12-Val, analogous to RGDS inhibition of His-12-Val binding to platelets. His-12-Val inhibited a minor component of transmembrane Ca2+ influx into ADP and thrombin-activated human platelets but had no effect on steady-state platelet 45Ca flux. These data indicate that ligand binding may exert a regulatory influence on transmembrane Ca2+ influx into activated platelets. The difference in inhibitory potency of the peptides studied may be related to differences in conformational changes in the glycoprotein IIb-IIIa complex induced by His-12-Val and RGDS, steric considerations, or differences in interactions with glycoprotein IIb Ca2+ binding domains. PMID- 2549027 TI - Tricarboxylate carrier of bovine liver mitochondria. Purification and reconstitution. AB - The tricarboxylate carrier of bovine liver mitochondria has been solubilized by Triton X-114 and purified by chromatography on hydroxylapatite and Silica Gel 60. The purified carrier could be visualized as a single band in polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate with Mr 37,000-38,000. The carrier, after reconstitution in phospholipid vesicles, catalyzed the exchange of [14C]citrate against citrate, malate, and threo-D8-isocitrate and was inhibited by the specific tricarboxylate carrier inhibitor 1,2,3 benzenetricarboxylic acid. PMID- 2549029 TI - Purification and properties of a protein inhibitor that inhibits phosphatase 2A activity when hydroxymethylglutaryl coenzyme A reductase is the substrate. AB - A heat-stable protein inhibitor of the hydroxymethylglutaryl-CoA reductase phosphatase 2A activity has been identified and purified to homogeneity, as judged by polyacrylamide gel electrophoresis. The apparent molecular mass was 20,000 Da. The protein lost its inhibitory properties when incubated with trypsin or treated with ethanol. The inhibitor protein does not inhibit type 1 phosphatase when either phosphorylase or hydroxymethylglutaryl-CoA reductase is the substrate. In contrast, this protein inhibitor inhibits the rat liver type 2A phosphatase activity when hydroxymethylglutaryl-CoA reductase is the substrate but not when phosphorylase a is the substrate. The inhibitor protein is not activated by incubation with ATP and cyclic AMP-dependent protein kinase and it is not phosphorylated by glycogen synthase kinase-3. These results, together with those of the kinetic experiments, suggest that the reductase phosphatase inhibitor is distinct from protein phosphatase inhibitor-1 and inhibitor-2. PMID- 2549028 TI - Multimeric structure of the tumor necrosis factor receptor of HeLa cells. AB - The tumor necrosis factor (TNF) receptor of HeLa cells was solubilized in Triton X-100 and characterized by gel filtration, affinity labeling, and ligand blotting studies. Receptors solubilized with Triton X-100 eluted in gel filtration as a major peak of Mr = 330,000 and retained high affinity binding (KD = 0.25 nM). Affinity labeling of soluble receptor/125I-TNF complexes using the reversible, bifunctional bis[2-(succinimidooxycarbonyl-oxy)ethyl] sulfone resulted in the formation of cross-linked species of Mr = 310,000, 150,000-175,000, 95,000, and 75,000. The formation of these complexes was competitively inhibited by unlabeled TNF. Partial reversal of cross-linking in these complexes and their analysis by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE) resolved 125I-TNF dimers cleaved from the 95,000 band and 125I-TNF monomer cleaved from the 75,000 band, providing evidence for a Mr approximately 60,000 subunit. In addition, the 95,000 and 75,000 bands were resolved as components of larger complexes (Mr = 150,000-175,000), which presumably contain two receptor subunits. The Mr 95,000 and 75,000 bands were also released from the Mr 310,000 complex by reduction with dithiothreitol, suggesting a role for disulfide bond stabilization. To investigate the association of the putative receptor subunits, Triton X-100 extracts from HeLa membranes were fractionated by SDS-PAGE without reduction and transferred electrophoretically to nylon membranes for TNF binding assays. Only two bands of Mr = 60,000 and 70,000 specifically bound TNF, and higher Mr binding activity was not observed. These results indicate that TNF receptors in HeLa cells are high molecular weight complexes containing Mr = 60,000 and 70,000 subunits each capable of binding TNF and that the complexes are primarily stabilized by non-covalent, hydrophobic interactions. PMID- 2549030 TI - The relative proton stoichiometries of the mitochondrial proton pumps are independent of the proton motive force. AB - Several different proton pumps were used to generate a proton motive force (delta p, proton motive force across the mitochondrial inner membrane) in isolated rat liver mitochondria, and the relationship between delta p and pump rate was investigated by titrating with various inhibitors of the pumps. It was found that this relationship was the same for mitochondria respiring on succinate irrespective of whether respiration was inhibited with malonate, antimycin or cyanide, indicating that the relationship was independent of the redox state of the respiratory chain. When delta p was generated by either the cytochrome bc1 complex, cytochrome oxidase, both together, or ATP hydrolysis (and transport), the reaction rates (in moles of electrons or ATP) were in the ratio of close to 3:1.5:1:1, respectively, at all accessible values of delta p. This suggests that the proton stoichiometries (H+/e and H+/ATP, where H+/e is the number of protons translocated vectorially across the inner membrane per electron transferred by the respiratory chain and H+/ATP is the number of protons translocated vectorially per ATP molecule hydrolyzed externally) were in the ratio of close to 1:2:3:3, respectively, at all values of delta p. Possible reasons for previous contradictory results are suggested. PMID- 2549031 TI - The sodium ion translocating oxaloacetate decarboxylase of Klebsiella pneumoniae. Sequence of the integral membrane-bound subunits beta and gamma. AB - The sequences upstream and downstream of the cloned gene for the alpha-subunit of the Na+ pump oxaloacetate decarboxylase of Klebsiella pneumonia were determined. An open reading frame in the upstream region was identified as the gene for the gamma-subunit, and an open reading frame in the downstream region represents the gene for the beta-subunit. The deduced primary structure of the gamma- and beta subunit was confirmed by protein sequencing of about 37 and 22%, respectively, of each polypeptide chain. The gene for the gamma-subunit has a GC content of 64% and codes for 83 amino acids. The protein is not processed at its amino terminus or at its carboxyl terminus. The gene for the beta-subunit has a GC content of 66% and codes for 327 amino acids. The protein contains a blocked aminoterminal methionine residue. Whether processing occurs at the carboxyl terminus is unknown. Hydropathy calculations defined one transmembrane helix in the amino terminal part of the gamma-subunit and a hydrophilic carboxyl-terminal part that is certainly not embedded within the lipid bilayer. A proline- and alanine-rich sequence in the carboxyl-terminal part may provide the protein with conformational flexibility. According to hydropathy and acrophilicity calculations, the secondary structure of the beta-subunit may be formed with 5 or 6 intramembrane helical segments. PMID- 2549032 TI - Multiple sources of 1,2-diacylglycerol in isolated rat pancreatic acini stimulated by cholecystokinin. Involvement of phosphatidylinositol bisphosphate and phosphatidylcholine hydrolysis. AB - Changes in the cellular content of 1,2-diacylglycerol (DAG) in isolated rat pancreatic acini in response to agonist stimulation were studied using a sensitive mass assay. When acini were stimulated by 10 nM COOH-terminal cholecystokinin-octapeptide (CCK8), the increase in DAG was biphasic, consisting of an early peak at 5 s and a second, larger, gradual increase that was maximal by 15 min. The basal level of DAG in acini was 1.04 nmol/mg of protein, which was increased to 1.24 nmol/mg of protein at 5 s and 2.76 nmol/mg of protein at 30 min. In comparison, the increase in DAG stimulated by 30 pM CCK8, a submaximal concentration for amylase release, was monophasic, increasing without an early peak but sustained to 60 min. Other Ca2+-mobilizing secretagogues such as carbamylcholine and bombesin increased DAG in acini, whereas vasoactive intestinal peptide, which acts to increase cAMP, had no effect. Phorbol ester and Ca2+ ionophore also stimulated DAG production. Analysis of the mass level of inositol 1,4,5-trisphosphate (1,4,5-IP3) showed that the generation of 1,4,5-IP3 stimulated by 10 nM CCK8 peaked at 5 s, a finding consistent with the early peak of DAG. The basal level was 4.7 pmol/mg of protein, which was increased to 144.6 pmol/mg of protein at 5 s by 10 nM CCK8. The levels of 1,4,5-IP3 then returned toward basal in contrast to the gradual and sustained increase of DAG. The dose dependencies of 1,4,5-IP3 and DAG formation at 5 s with respect to CCK8 were almost identical. This suggests that phosphatidylinositol 4,5-bisphosphate hydrolysis is a major source of the early increase in DAG but not of the sustained increase in DAG. Therefore, a possible contribution of phosphatidylcholine hydrolysis to DAG formation was examined utilizing acini prelabeled with [3H]choline. CCK8 (1 nM) maximally increased [3H]choline metabolite release by 133% of control at 30 min. Separation of these metabolites by thin layer chromatography showed that the products of CCK8-stimulated release were almost entirely phosphorylcholine, indicating the activation of a phospholipase C specific for phosphatidylcholine. By comparison, 1 nM CCK8 stimulated [3H]ethanolamine metabolite release from [3H]ethanolamine-labeled acini by only 22% of control. These data suggest that CCK stimulates both phosphatidylinositol 4,5-bisphosphate and phosphatidylcholine hydrolysis; the latter may contribute to the sustained generation of DAG and hence the maintained activation of protein kinase C. PMID- 2549033 TI - Muscarinic receptor activation of phosphatidylcholine hydrolysis. Relationship to phosphoinositide hydrolysis and diacylglycerol metabolism. AB - We examined the relationship between phosphatidylcholine (PC) hydrolysis, phosphoinositide hydrolysis, and diacylglycerol (DAG) formation in response to muscarinic acetylcholine receptor (mAChR) stimulation in 1321N1 astrocytoma cells. Carbachol increases the release of [3H]choline and [3H]phosphorylcholine ([3H]Pchol) from cells containing [3H]choline-labeled PC. The production of Pchol is rapid and transient, while choline production continues for at least 30 min. mAChR-stimulated release of Pchol is reduced in cells that have been depleted of intracellular Ca2+ stores by ionomycin pretreatment, whereas choline release is unaffected by this pretreatment. Phorbol 12-myristate 13-acetate (PMA) increases the release of choline, but not Pchol, from 1321N1 cells, and down-regulation of protein kinase C blocks the ability of carbachol to stimulate choline production. Taken together, these results suggest that Ca2+ mobilization is involved in mAChR mediated hydrolysis of PC by a phospholipase C, whereas protein kinase C activation is required for mAChR-stimulated hydrolysis of PC by a phospholipase D. Both carbachol and PMA rapidly increase the formation of [3H]phosphatidic acid ([3H]PA) in cells containing [3H]myristate-labeled PC. [3H]Diacylglycerol ([3H]DAG) levels increase more slowly, suggesting that the predominant pathway for PC hydrolysis is via phospholipase D. When cells are labeled with [3H]myristate and [14C]arachidonate such that there is a much greater 3H/14C ratio in PC compared with the phosphoinositides, the 3H/14C ratio in DAG and PA increases with PMA treatment but decreases in response to carbachol. By analyzing the increase in 3H versus 14C in DAG, we estimate that the DAG that is formed in response to PMA arises largely from PC. Muscarinic receptor activation also causes formation of DAG from PC, but approximately 20% of carbachol-stimulated DAG appears to arise from hydrolysis of the phosphoinositides. PMID- 2549034 TI - Human deoxycytidine kinase. Sequence of cDNA clones and analysis of expression in cell lines with and without enzyme activity. AB - Deoxycytidine kinase (dC kinase) is the rate-limiting enzyme in the anabolism of important anticancer and retroviral nucleoside derivatives. Its activity is often decreased in resistance to these drugs. To analyze the structure, function, and control of this clinically important enzyme we isolated 15 cDNA clones for human deoxycytidine kinase from lambda gt11 thymus and Molt 4 libraries. Four clones were sequenced. The largest clone is 2.9 kilobases and codes for a 626-amino acid open reading frame. The DNA and deduced amino acid sequence of the human dC kinase clones are homologous with a previously unidentified murine cDNA clone p3.4J (EMBL:MM34j) reported to be related to granulocyte-macrophage colony stimulating factor. Deoxycytidine kinase also has cysteine-rich regions that are homologous with thioredoxin, the beta subunit of prolyl 4-hydroxylase, phosphoinositide-specific phospholipase C, thyroid hormone-binding protein, and protein disulfide isomerase. No differences were seen in the amount and size of deoxycytidine kinase protein and mRNA between CCRF/CEM and L1210 leukemic cell lines that express and do not express enzyme activity. Genomic restriction fragments were similar between the active and inactive CCRF/CEM cell lines. These data suggest that the cells deficient in dC kinase activity have a small defect in the structural gene. PMID- 2549035 TI - Sequence of the cloned Escherichia coli K1 CMP-N-acetylneuraminic acid synthetase gene. AB - The Escherichia coli CMP-N-acetylneuraminic acid (CMP-NeuAc) synthetase gene is located on a 3.3-kilobase (kb) HindIII fragment of the plasmid pSR23 which contains the genes for K1 capsule production (Vann, W. F., Silver, R. P., Abeijon, C., Chang, K., Aaronson, W., Sutton, A., Finn, C. W., Lindner, W., and Kotsatos, M. (1987) J. Biol. Chem. 262, 17556-17562). The CMP-NeuAc synthetase gene expression was increased 10-30-fold by cloning of a 2.7-kb EcoRI-HindIII fragment onto the vector pKK223-3 containing the tac promoter. The complete nucleotide sequence of the gene encoding CMP-NeuAc synthetase was determined from progressive deletions generated by selective digestion of M13 clones containing the 2.7-kb fragment. CMP-NeuAc synthetase is located near the EcoRI site on this fragment as indicated by the detection of an open reading frame encoding a 49,000 dalton polypeptide. The amino- and carboxyl-terminal sequences of the encoded protein were confirmed by sequencing of peptides cleaved from both ends of the purified enzyme. The nucleotide deduced amino acid sequence was confirmed by sequencing several tryptic peptides of purified enzyme. The molecular weight is consistent with that determined from sodium dodecyl sulfate-gel electrophoresis. Gel filtration and ultracentrifugation experiments under nondenaturing conditions suggest that the enzyme is active as a 49,000-dalton monomer but may form aggregates. PMID- 2549036 TI - Purification and in vitro DNA-binding specificity of the Bacillus subtilis phage phi 105 repressor. AB - The Bacillus subtilis phage phi 105 repressor, a lambda repressor-like transcriptional regulatory protein, was overproduced in Escherichia coli and purified to near homogeneity in order to examine its in vitro DNA-binding properties. The active form of repressor appears to be a tetramer. DNase I protection experiments demonstrate that repressor can specifically bind to six distinct sites, all located within the phi 105 EcoRI-F immunity region (immF). Three of these sites had been identified earlier as functional operators by genetic analysis. They share a common 14-base pair, asymmetric "core" sequence, 5'-GACGGAAATACAAG-3', termed OR. The three additional sites show significant homology with OR. For an individual binding site, hydroxyl-radical footprinting reveals symmetrical repressor-DNA interactions established via one side of the helix. Dimethyl sulfate protection indicates that guanines at the conserved OR base pair positions 1, 3, and 4 may participate in sequence-specific interactions with repressor in agreement with a previously proposed recognition model. However, since the OR sequence is not symmetrical with respect to this GNCG motif, at present it remains difficult to completely understand the molecular basis of this interaction. PMID- 2549038 TI - Identification of an aldehyde dehydrogenase in the microsomes of human polymorphonuclear leukocytes that metabolizes 20-aldehyde leukotriene B4. AB - We have previously reported that cytochrome P-450LTB in the microsomes of human polymorphonuclear leukocytes (PMN) catalyzes three omega-oxidations of leukotriene B4 (LTB4), leading to the sequential formation of 20-OH-LTB4, 20-CHO LTB4, and 20-COOH-LTB4 (Soberman, R.J., Sutyak, J.P., Okita, R.T., Wendelborn, D.F., Roberts, L.J., II, and Austen, K. F. (1988) J. Biol. Chem. 263, 7996-8002). The identification of the novel final intermediate, 20-CHO-LTB4, allowed direct analysis of its metabolism by PMN microsomes in the presence of adenine nucleotide cofactors. Microsomes in the presence of 100 microM NAD+ or 100 microM NADP+ converted 1.0 microM 20-CHO-LTB4 to 20-COOH-LTB4 with a Km of 2.4 +/- 0.8 microM (mean +/- S.E., n = 4) and a Vmax of 813.9 +/- 136.6 pmol.min-1.mg-1, for NAD+, as compared to 0.12 microM and 5.0 pmol.min-1.mg-1 (n = 2) for NADPH as a cofactor. The conversion of 1.0 microM of 20-CHO-LTB4 to 20-COOH-LTB4 in the presence of saturating concentrations (1.0 mM) of both NAD+ and NADP+ was not greater than the reaction in the presence of 1.0 mM of each cofactor separately, indicating that NAD+ and NADP+ were cofactors for the same enzyme. Antibody to cytochrome P-450 reductase did not inhibit the conversion of 20-CHO-LTB4 to 20 COOH-LTB4. When 1.0 microM 20-OH-LTB4 was added to microsomes in the presence of NADPH, approximately three-fourths of the product formed (63.7 +/- 5.1 pmol; mean +/- S.E., n = 3) was 20-CHO-LTB4 and approximately one-fourth (21.3 +/- 3.9 pmol; mean +/- S.E., n = 3) was 20-COOH-LTB4. In the presence of both NADPH and NAD+, only 20-COOH-LTB4 (85.5 +/- 9.9 pmol; mean +/- S.E., n = 3) was formed. PMN microsomes also contain an NADH-dependent aldehyde reductase which converts 20 CHO-LTB4 to 20-OH-LTB4, a member of the LTB4 family of molecules with biological activity. Based upon kinetic, cofactor and inhibition data, microsomal aldehyde dehydrogenase preferentially regulates the final and irreversible inactivation step in the LTB4 metabolic sequence. PMID- 2549037 TI - Purification and characterization of recombinant human parathyroid hormone related protein. AB - Full-length human parathyroid hormone-related protein (PTHrP-(1-141] as well as a carboxyl-terminal shortened form (PTHrP-(1-108] have been expressed from recombinant DNA-derived clones. These proteins were expressed in Escherichia coli as fusion proteins so that cyanogen bromide cleavage yields the desired product. Both proteins were purified and then characterized by sodium dodecyl sulfate gel electrophoresis, amino-terminal amino acid sequencing, peptide mapping, and mass spectral analysis. Recombinant PTHrP-(1-141), PTHrP-(1-108), synthetic PTHrP-(1 34), and naturally derived PTHrP are all equipotent in the stimulation of cyclic AMP levels in the osteoblast-like cell line UMR 106-01. However, PTHrP-(1-141) and -(1-108) are two to four times more active than PTHrP-(1-34) in the stimulation of plasminogen activator activity from this cell line. PTHrP-(1-141) reacts equipotently with PTHrP-(1-34) in a radioimmunoassay using an antiserum prepared against PTHrP-(1-34). PTHrP-(1-141), -(1-108), and -(1-84) were used as PTHrP-specific mobility standards on sodium dodecyl sulfate gel electrophoresis to determine the approximate length of two forms of naturally derived PTHrP. The data show that PTHrP purified from the lung tumor cell line BEN contains a major form of about 108 amino acids and another form of about 141 amino acids. PMID- 2549039 TI - Effector coupling mechanisms of the cloned 5-HT1A receptor. AB - The signal transduction pathways of the cloned human 5-HT1A receptor have been examined in two mammalian cell lines transiently (COS-7) or permanently (HeLa) expressing this receptor gene. In both systems, 5-hydroxytryptamine (5-HT, serotonin) mediated a marked inhibition of beta 2-adrenergic agonist-stimulated (80% inhibition in COS-7 cells) or forskolin-stimulated cAMP formation (up to 90% inhibition in HeLa cells). This serotonin effect (EC50 = 20 nM) could be competitively antagonized by metitepine and spiperone (Ki = 81 and 31 nM, respectively) and could also be blocked by pretreatment of cells with pertussis toxin. In both cell types, 5-HT failed to stimulate adenylyl cyclase through the expressed receptors. In HeLa cells, 5-HT also stimulated phospholipase C (approximately 40-75% stimulation of formation of inositol phosphates). Again, this effect was inhibited by metitepine. However, the EC50 of 5-HT was considerably higher (approximately 3.2 microM) than that found for inhibition of adenylyl cyclase. Both pathways were demonstrated to be similarly affected by pertussis toxin. These findings indicate that like the M2 and M3 muscarinic cholinergic receptors, the 5-HT1A receptor can couple to multiple transduction pathways with varying efficiencies via pertussis toxin-sensitive G-proteins. The lack of stimulation of cAMP formation by this 5-HT1A receptor may suggest the existence of another pharmacologically closely related receptor. PMID- 2549041 TI - Phorbol ester transiently increases topoisomerase I mRNA levels in human skin fibroblasts. AB - Exposure of human skin fibroblasts to phorbol 12-myristate 13-acetate (PMA) results in a transient increase in the level of topoisomerase I mRNA. A maximal increase (about 10-fold) in the level of topoisomerase I mRNA was observed 3 h after adding 200 nM PMA, but a decrease to the basal level was observed within 12 h of PMA treatment. The lowest PMA concentration to give an observable induction of topoisomerase I gene expression was found to be 20 nM. In addition, the induction of topoisomerase I gene expression by 20 nM PMA pulse treatment for 10 min followed by incubation for an additional 3 h attained the same level of induction seen with 3 h of continuous exposure to 20 nM PMA. This observation suggests that once the signal for protein kinase C activation is transduced, continuous exposure to PMA is not necessary for the maximal effect. To gain insight into the mechanism by which PMA stimulates topoisomerase I gene expression, cells were treated with PMA, cycloheximide, and actinomycin D, either alone or in various combinations. The results show that actinomycin D, but not cycloheximide, specifically abolishes the stimulatory effect of PMA, suggesting that PMA affects topoisomerase I gene expression at the transcriptional level. PMID- 2549040 TI - Zwitterionic and anionic forms of a serotonin analog as transport substrates. AB - 4,6-Difluoroserotonin, a serotonin analog with an acidic 5-hydroxyl proton (pK alpha = 7.97) relative to serotonin (pK alpha = 10.73), was tested as a substrate for the biogenic amine transporter of bovine chromaffin granules and the plasma membrane serotonin transporter of human blood platelets. The platelet serotonin transporter transports this analog with identical rates as those for serotonin, both at pH 6.7, where the hydroxyl group is predominantly protonated and at pH 9, where it is largely dissociated. In contrast, the chromaffin granule biogenic amine transporter prefers the form of 4,6-difluoroserotonin with a protonated 5 hydroxyl group. Thus, the KM for 4,6-difluoroserotonin increases, and Vmax decreases (relative to the values for serotonin) as the pH increases from 7 to 9. This effect may reflect a specific requirement for the protonated hydroxyl group in substrate translocation, as opposed to binding, since the KI for 4,6 difluoroserotonin inhibition of serotonin transport is the same as the KM for serotonin from pH 7 to 9. PMID- 2549042 TI - Two different cell types have different major receptors for human tumor necrosis factor (TNF alpha). AB - The receptors for tumor necrosis factor alpha (TNF alpha) were analyzed on myeloid cells (HL60, U937, K562, and freshly isolated blood monocytes) and on cells of epithelial origin (MCF7, HEp2 and HeLa cells), by use of radiolabeled TNF alpha and cross-linking experiments. Both cell types had high but slightly different affinities for TNF alpha. The myeloid cells had major cross-linked products of 98-100 kDa, which were similar in their N-linked glycosylation, whereas the cells of epithelial origin contained a major cross-linked product of 75 kDa, a second product of 95 kDa. The major receptors of both cell types (studied mostly with HL60 and HEp2 cells) are different proteins because (a) their apparent molecular masses were different and no evidence was obtained for cell-specific proteases, which could generate the differently sized receptors from one common receptor molecule; (b) anti-receptor antibodies, which precipitated the 95- and 75-kDa products, did not precipitate the 100-kDa cross linked complex; (c) the native TNF alpha-receptor complexes had different proteolytic fingerprints; (d) the tryptic fragments differed in their association with the cell membrane vesicles; (e) the receptors differed in their degree of N linked glycosylation; and (f) O-linked glycosylation was found on the major receptor of HL60 but not of HEp2 cells. In addition, myeloid cells may also contain a small amount of the HEp2-type of TNF alpha receptor. We suggest that at least two different receptors for TNF alpha exist. PMID- 2549043 TI - Protein phosphorylation associated with synergistic stimulation of neutrophils. AB - Neutrophils treated with optimal amounts of tumor promoters that activate protein kinase C (e.g. mezerein) release large quantities of superoxide (O2-) and exhibit an intense phosphorylation of two proteins with molecular masses of approximately 47 and 49 kDa. These cells can also be stimulated synergistically to release a comparable amount of O2-. This involves treatment with a suboptimal amount of a tumor promoter and an agent capable of elevating cellular Ca2+. Neutrophils treated in the former fashion exhibit a redistribution of the activity of protein kinase C from a soluble to a particulate fraction that is stable in the presence of Ca2+ chelators, whereas cells stimulated synergistically do not do so to an appreciable extent (Badwey, J. A., Robinson, J. M., Horn, W., Soberman, R. J., Karnovsky, M. J., and Karnovsky, M. L. (1988) J. Biol. Chem. 263, 2779-2786). In this paper, we report that neutrophils stimulated synergistically do exhibit a significant incorporation of 32P into the 47-kDa protein, but with little labeling of the 49-kDa species. This labeling of the 47-kDa protein was greater than the sum of those observed with each agent added separately but was less than that observed in cells stimulated with optimal amounts of tumor promoters alone. An inhibitor of protein kinase C (1-(5-isoquinolinylsulfonyl)-2-methylpiperazine) blocked O2- release and the phosphorylation of the 47-kDa protein under all conditions of stimulation mentioned, whereas an inhibitor of cyclic nucleotide dependent kinases had no effect on these phenomena. Thus, labeling of the 47-kDa protein can occur in the absence of a "tight" translocation of protein kinase C to membrane and was always observed during synergy. The data support a role for protein kinase C and the 47-kDa phosphoprotein in the synergistic stimulation of neutrophils. PMID- 2549044 TI - Paradoxical effects of retinal in neutrophil stimulation. AB - Retinal stimulates the activity of phospholipase C and superoxide (O2-) release in neutrophils. The latter response is comparable in magnitude to that observed when phorbol 12-myristate 13-acetate (PMA) is the stimulating agent. Cells treated with retinal, however, do not undergo degranulation, nor do they exhibit the formation of intracellular vesicles, as is commonly observed with other agents (e.g. Lochner, J. E., Badwey, J. A., Horn, W., and Karnovsky, M. L. (1986) Proc. Natl. Acad. Sci. U. S. A. 83, 7673-7677). Retinal promotes redistribution of the activity of protein kinase C from a soluble to a particulate fraction in neutrophils, and this redistribution precedes O2- release. Superoxide release stimulated with retinal, however, is largely insensitive to inhibitors of protein kinase C (1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7); staurosporine). These compounds substantially block both O2- release and the phosphorylation of two proteins with molecular masses of about 47 and 49 kDa when the stimulus is PMA. The data indicate that retinal and PMA elicit the formation of active protein kinase C complexes of different natures, or that the mechanism of stimulation of O2- release by retinal does not involve this kinase. The significance of these observations to the common use of retinoids as inhibitors of protein kinase C is discussed. PMID- 2549045 TI - Alteration of the phospholipid composition of Escherichia coli through genetic manipulation. AB - In order to study the function of individual phospholipids, we have constructed a strain of Escherichia coli in which the ratio of phosphatidylethanolamine to phosphatidylglycerol plus cardiolipin can be regulated. In this strain (HDL1001) the normal expression of the phosphatidylglycerophosphate synthase does not occur due to the presence of the pgsA30 allele (Heacock, P. N., and Dowhan, W. (1987) J. Biol. Chem. 262, 13044-13049). A second chromosomal copy of the pgsA gene is fused to the lacOP region in single copy within the lac operon. Strain HDL1001 is absolutely dependent for growth on an inducer of the lac operon. In addition, the level of the pgsA gene product, the content of the two major acidic phospholipids, and the growth rate are dependent on the level of inducer in the growth medium. Cells remain viable in the absence of inducer as evidenced by a rapid return to normal growth after the readdition of inducer. The growth rate and phospholipid composition are affected only after the level of phosphatidylglycerophosphate synthase drops below about 15% of normal levels; both phosphatidic acid and (d)CDP-diacylglycerol also begin to increase to significant levels. At the point of cell arrest the level of the major acidic phospholipids is reduced by about 90% of wild type levels. PMID- 2549046 TI - Quantitative changes in polyphosphoinositides 1,2-diacylglycerol and inositol 1,4,5-trisphosphate by platelet-derived growth factor and prostaglandin F2 alpha. AB - We developed a novel method to quantify trace amounts of phosphatidylinositol 4 phosphate (PIP) and phosphatidylinositol 4,5-bisphosphate (PIP2) using antibodies against PIP and PIP2. With this method, polyphosphoinositides can be measured in the range from 20 to 500 pmol. We applied the method to quantify changes in PIP and PIP2 levels in Balb/c/3T3 cells stimulated by platelet-derived growth factor (PDGF) and prostaglandin F2 alpha (PGF2 alpha), growth factors that stimulate the hydrolysis of PIP and PIP2. PIP2 content decreased rapidly to about 60% of control within 1 min while PIP content decreased gradually but significantly to 60% (PDGF) or 70% (PGF2 alpha) of control. Simultaneously we measured the mass levels of inositol 1,4,5-trisphosphate and 1,2-diacylglycerol (DG). Inositol 1,4,5-trisphosphate levels rapidly increased and reached a maximum at 30 s after PDGF or PGF2 alpha stimulation and then decreased to the control level within 2 min. On the other hand, DG formation showed biphasic changes. In the first phase, DG rapidly accumulated and reached a maximum at 30 s after PDGF or PGF2 alpha stimulation and then quickly decreased. In the second phase, DG accumulated gradually, but very markedly, 2 min after PDGF or PGF2 alpha stimulation. Considering the changes in PIP2, DG in the first phase seems to be derived mainly from PIP2 while most of the DG in the second phase derived from other lipids. PMID- 2549047 TI - Adenosine-5'-phosphosulfate kinase from Escherichia coli K12. Purification, characterization, and identification of a phosphorylated enzyme intermediate. AB - Adenosine-5'-phosphosulfate kinase (ATP:adenylylsulfate 3'-phosphotransferase), the second enzyme in the pathway of sulfate activation, has been purified (approximately 300-fold) to homogeneity from an Escherichia coli K12 strain, which overproduces the enzyme activity (approximately 100-fold). The purified enzyme has a specific activity of 153 mumol of 3'-phosphoadenosine 5' phosphosulfate (PAPS) formed/min/mg of protein at 25 degrees C. The enzyme is remarkably efficient with a Vmax/Km(APS) of greater than 10(8) M-1 s-1, indicating that at physiologically low substrate concentrations the reaction is essentially diffusion limited. Upon incubation with MgATP a phosphorylated enzyme is formed; the isolated phosphorylated enzyme can transfer its phosphoryl group to adenosine 5'-phosphosulfate (APS) to form PAPS or to ADP to form ATP. The phosphorylated enzyme exists as a dimer of identical 21-kilodalton subunits, while the dephosphorylated form primarily exists as a tetramer. Divalent cations are required for activity with Mg(II), Mn(II), Co(II), and Cd(II) activating. Studies of the divalent metal-dependent stereoselectivity for the alpha- and beta phosphorothioate derivatives of ATP indicate metal coordination to at least the alpha-phosphoryl group of the nucleotide. Steady state kinetic studies of the reverse reaction indicate a sequential mechanism, with a rapid equilibrium ordered binding of MgADP before PAPS. In the forward direction APS is a potent substrate inhibitor, competitive with ATP, complicating kinetic studies. The primary kinetic mechanism in the forward direction is sequential. Product inhibition studies at high concentrations of APS suggest an ordered kinetic mechanism with MgATP binding before APS. At submicromolar concentrations of APS, product inhibition by both MgADP and PAPS is more complex and is not consistent with a solely ordered sequential mechanism. The formation of a phosphorylated enzyme capable of transferring its phosphoryl group to APS or to MgADP suggests that a ping-pong pathway in which the rate of MgADP dissociation is comparable to the rate of APS binding might contribute at very low concentrations of APS. The substrate inhibition by APS is consistent with APS binding to the enzyme, to form a dead-end E.APS complex. PMID- 2549048 TI - Parallel effects of signal peptide hydrophobic core modifications on co translational translocation and post-translational cleavage by purified signal peptidase. AB - The length of the hydrophobic core of the bovine parathyroid hormone signal peptide was modified by in vitro mutagenesis. Extension of the hydrophobic core by three amino acids at the NH2-terminal end had little effect on the proteolytic processing of the signal peptide by microsomal membranes. Deletion of 6 of the 12 amino acids in the core eliminated translocation and processing of the modified protein. Deletion of pairs of amino acids across the core resulted in position dependent inhibition of signal activity unrelated to hydrophobicity but inversely related to the hydrophobic moments of the modified cores. Deletions in the NH2 terminal region of the core were strongly inhibitory for proteolytic processing whereas deletions in the COOH-terminal region had no effect or increased processing when assessed either co-translationally with microsomal membranes or post-translationally with purified hen oviduct signal peptidase. Deletion of cysteine 18 and alanine 19 increased processing, but deletion of cysteine alone or substitution of leucine for cysteine did not increase processing more than deletion of both residues at 18 and 19. Translations of the translocation defective mutants with pairs of amino acids deleted in a wheat germ system were inhibited by addition of exogenous signal recognition particle suggesting that interactions of the modified signal peptides with signal recognition particle were normal. The position-dependent effects of the hydrophobic core modifications indicate that structural properties of the core in addition to hydrophobicity are important for signal activity. The parallel effects of the modifications on co translational translocation and post-translational processing by purified signal peptidase suggest that proteins in the signal peptidase complex might be part of, or intimately associated with, membrane proteins involved in the translocation. A model is proposed in which the NH2-terminal region of the hydrophobic core binds to one subunit of the signal peptidase while the other subunit catalyzes the cleavage. PMID- 2549049 TI - Proteolytic activity associated with the nuclear scaffold. The effect of self digestion on lamins. AB - The inner aspect of the nuclear envelope is supported by a peripheral framework called the nuclear scaffold, which consists of both structural and functional proteins. Its major structural components, lamins A-C, form a highly polymerized and insoluble fibrous matrix during interphase of the cell cycle. Functional constituents of the scaffold include the 46-kDa nucleoside triphosphatase which is thought to participate in nucleocytoplasmic transport of mRNA. This 46-kDa component shares an amino-terminal sequence with lamins A and C, indicating that proteolytic remodeling of the nuclear scaffold may contribute to the generation of nucleoside triphosphatase activity (Clawson, G. A., Lackey, A., and Tokes, Z. A. (1988) Exp. Cell Res. 176, 180-186). We report here that neutral protease activity intimately associated with the nuclear scaffold is also a functional constituent. This activity has a considerable selectivity for lamins as shown by self-digestion of scaffold preparations, and it may participate in the remodeling of the nuclear scaffold after treatment with carcinogens. PMID- 2549051 TI - Fibrinogen sialic acid residues are low affinity calcium-binding sites that influence fibrin assembly. AB - Calcium ions occupy low (n congruent to 10; Kd congruent to 1 mM) and high (n = 3; Kd congruent to 1 microM) affinity sites on fibrinogen and facilitate fibrin monomer polymerization. We have previously localized two of the three high affinity Ca2+ sites to gamma 311-gamma 336. However, optimal enhancement of fibrin monomer polymerization occurs only at physiological millimolar Ca2+ concentrations which are two orders of magnitude higher than the concentration required for occupancy of the high affinity Ca2+-binding sites. In this study, we show that removal of fibrinogen sialic acid residues results in loss of low affinity Ca2+-binding sites. Clotting of asialofibrinogen appears to be Ca2+ independent and results in fiber bundles thicker in diameter than normal fibrin bundles as determined by turbidometry and scanning and transmission electron microscopy. By using a Ca2+-sensitive electrode, free sialic acid is shown to bind Ca2+ (Kd congruent to 1 mM). These observations suggest that the high affinity fibrinogen D-domain Ca2+-binding sites may play a role in the tertiary structure of the D-domain, whereas, sialic acid residues are low affinity sites whose occupancy by Ca2+ at physiological calcium concentration facilitates fibrin polymerization. PMID- 2549050 TI - Characterization of the spoT gene of Escherichia coli. AB - The Escherichia coli spoT gene encodes a guanosine-3',5'-bispyrophosphate (ppGpp) 3'-pyrophosphohydrolase known to be responsible for cellular (ppGpp) degradation. The DNA sequence of the spoT region is presented. The spoT gene is deduced to be 702 codons long, with a probable UUG initiation codon, and a deduced mass of 79,342 daltons. Two spoT mutations (spoT202 and spoT203) have been localized to an open reading frame by complementation of function as well as by genetic marker rescue. The ability to overexpress the spoT gene is limited, but enough ppGppase activity can be made to reverse ppGpp accumulation during the stringent response to amino acid starvation. The spoT gene is located within a larger spo operon and is flanked by two smaller genes. The first gene in the operon encodes omega, a protein that copurifies with RNA polymerase (Gentry, D. R., and Burgess, R. R. (1986) Gene (Amst.) 48, 33-40). The spoT gene is the second gene in the operon; it is followed by a third open reading frame deduced to encode a protein with a mass of 25,343 daltons. Insertion of a kanamycin resistance gene in the omega gene reduces spoT gene expression as judged by lowered ppGppase activity, relA dependent reduction of growth rate, and abolition of spoT mutant complementation activity. These effects are reversed by expression of the spoT gene, but not the omega gene, in trans. Transcription of the spo operon occurs in a clockwise direction on the E. coli chromosome and is probably directed by at least two promoters. PMID- 2549052 TI - Myosin heavy chain kinase from developed Dictyostelium cells. Purification and characterization. AB - We purified to homogeneity the Dictyostelium discoideum myosin heavy chain kinase that is implicated in the heavy chain phosphorylation increases that occur during chemotaxis. The kinase is initially found in the insoluble fraction of developed cells. The major purification step was achieved by affinity chromatography using a tail fragment of Dictyostelium myosin (LMM58) expressed in Escherichia coli (De Lozanne, A., Berlot, C. H., Leinwand, L. A., and Spudich, J. A. (1988) J. Cell Biol. 105, 2990-3005). The kinase has an apparent molecular weight of 84,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The apparent native molecular weight by gel filtration is 240,000. The kinase catalyzes phosphorylation of myosin heavy chain or LMM58 with similar kinetics, and the extent of phosphorylation for both is 4 mol of phosphate/mol. With both substrates the Vmax is about 18 mumol/min/mg and the Km is 15 microM. The myosin heavy chain kinase is specific to Dictyostelium myosin heavy chain, and the phosphorylated amino acid is threonine. The kinase undergoes autophosphorylation. Each mole of kinase subunit incorporates about 20 mol of phosphates. Phosphorylation of myosin by this kinase inhibits myosin thick filament formation, suggesting that the kinase plays a role in the regulation of myosin assembly. PMID- 2549053 TI - In vivo phosphorylation of the 170-kDa form of eukaryotic DNA topoisomerase II. Cell cycle analysis. AB - We have examined the level of incorporation of 32P into DNA topoisomerase II in vivo in chicken lymphoblastoid cells that were fractionated into the various cell cycle phases by centrifugal elutriation. We find that topoisomerase II is phosphorylated in vivo, with the level of incorporation being approximately 3.5 fold higher in the G2 + M fraction than earlier in the cell cycle. Our antibody studies have revealed that topoisomerase II antigen exists as a number of discrete polypeptide species in these cells. Of these, the 170-kDa intact polypeptide is phosphorylated approximately 4.5-fold more than several antigenic fragments that actually comprise the bulk of the topoisomerase II antigen in these cells at mitosis. Phosphorylation of the 170-kDa form of the enzyme may be involved in activation of the enzyme for its role in the disjunction of sister chromatids at anaphase. PMID- 2549054 TI - A novel cell adhesive protein engineered by insertion of the Arg-Gly-Asp-Ser tetrapeptide. AB - A tetrapeptide Arg-Gly-Asp-Ser (RGDS) has been shown to be a versatile cell recognition signal of extracellular matrix components for the interaction with cells. We introduced the RGDS tetrapeptide into a truncated form of protein A, a staphylococcal immunoglobulin-binding protein, by inserting an oligonucleotide cassette encoding the tetrapeptide into the coding region of the protein A expression vector pRIT2T. The mutagenized protein was capable of not only binding to immunoglobulin G but also mediating cell attachment and spreading onto an inert substrate. Cell adhesion mediated by the mutagenized protein was inhibitable by a synthetic peptide Gly-Arg-Gly-Asp-Ser but not by a related peptide Gly-Arg-Gly-Glu-Ser, confirming that the inserted RGDS tetrapeptide served as a recognition signal for cell adhesion. Furthermore, the RGDS containing protein was capable of adhering cells onto an immunoglobulin-coated surface which could not by itself support cell adhesion. Thus, the cell adhesive and immunoglobulin binding activities of the mutagenized protein appear to function coordinately. The protocol described here is essentially applicable to any protein and, therefore, provides a general principle in tailoring novel multifunctional proteins having cell adhesive activity. PMID- 2549056 TI - The kinetics of steroidogenesis activator polypeptide in the rat adrenal cortex. Effects of adrenocorticotropin, cyclic adenosine 3':5'-monophosphate, cycloheximide, and circadian rhythm. AB - The behavior of steroidogenesis activator polypeptide (SAP), a recently described modulator of cholesterol side-chain cleavage activity (Pedersen, R. C., and Brownie, A. C. (1987) Science 236, 188-190), was investigated in rat adrenocortical cells using a specific radioimmunoassay. In response to a maximal dose of adrenocorticotropic hormone (ACTH) (1 nM) or of 8-Br-cAMP (1 mM), an increase in intracellular SAP begins rapidly (less than 1 min) and reaches half maximal and maximal levels (16-fold greater than basal) at 3 and 15 min, respectively. A plateau at this maximal concentration of SAP is then maintained. The levels of intracellular SAP content and of corticosterone output exhibit a similar dose-dependent response to ACTH (EC50 = 25 and 30 pM, respectively). Treatment of ACTH-stimulated cells with cycloheximide reverses the rise in SAP (t1/2 congruent to 5-7 min). In vivo the SAP content of adrenals from quiescent rats is concordant with the circadian rhythm of the pituitary-adrenal axis; at the apex (1800 h), adrenal SAP is 13-fold higher than at the nadir (0800 h), paralleling 2- and 7-fold variations in cholesterol side-chain cleavage activity and serum corticosterone levels, respectively. At both time points, SAP levels rise in response to stress. Of the rat tissues examined, only the major steroid forming organs (adrenal cortex and gonads) had significant levels of immunoreactive, cAMP-responsive SAP, while cAMP-unresponsive immunoreactivity was also detectable in the thymus, spleen, and brain. Considered together with the biological activity previously demonstrated for SAP in vitro, these data are consistent with its role as a cAMP-dependent, cycloheximide-sensitive modulator of steroid biosynthesis. PMID- 2549057 TI - Effects of tryptophan and pH on the kinetics of superoxide radical binding to indoleamine 2,3-dioxygenase studied by pulse radiolysis. AB - The reaction of superoxide radical (O2-) with the heme protein indoleamine 2,3 dioxygenase has been investigated by the use of pulse radiolysis. In the absence of the substrate tryptophan (Trp), the ferric enzyme reacted quantitatively with O2- to form the oxygenated enzyme. The rate constant for the reaction (8.0 x 10(6) M-1 s-1 at pH 7.0) increased with a decrease in pH. In the presence of low concentrations of L-Trp (approximately 50 microM), under which the catalytic site of the ferric enzyme is greater than 99% Trp-free at pH 7.0, the only spectral species observed upon O2- binding was L-Trp-bound oxygenated enzyme, the ternary complex. This suggests that under the conditions employed O2- binds first to the ferric enzyme to form the oxygenated enzyme and is followed by rapid binding of L Trp. It was also found that absorbance changes (delta A) for the enzyme after the pulse were significantly decreased when an increased L-Trp concentration was employed. A 50% decrease in delta A was caused with approximately 50 microM L-Trp at pH 7.0. Similar results were also observed with other indole derivatives with decreasing delta A values in the order of indole, 3-indoleethanol, alpha-methyl DL-Trp, and D-Trp. These results suggest that there exists a binding site for these compounds in the dioxygenase different from the catalytic site for Trp and, most significantly, that binding of Trp to the effector binding site of the ferric enzyme markedly inhibits its reaction with O2-. PMID- 2549055 TI - Enhancement of adenosine A2 and prostaglandin E1 receptor-mediated cAMP generation by prior exposure of Swiss 3T3 fibroblasts to Ca2+-mobilizing receptor agonists or phorbol ester. Activation of protein kinase C triggers increases in the receptor density in cell membranes. AB - Production of cAMP in response to adenosine A2 or prostaglandin E1 receptor stimulation was, but the production induced by a beta-adrenergic agonist or forskolin was not, enhanced by prior exposure of Swiss 3T3 fibroblasts to agonists of Ca2+-mobilizing receptors or phorbol ester for 3 h. The enhancement reflected potentiation of the receptor-coupled activation of adenylate cyclase and the 2-fold increase in the adenosine A2 receptor number in membranes under these conditions. No enhancement was observed, however, when the medium used for the prior exposure was further supplemented with 1-(5-isoquinolinesulfonyl)-2 methylpiperazine (H-7) or staurosporin, inhibitors of protein kinase C, neither of which affected the cAMP responses of the nonexposed cells. It is very likely, therefore, that activation of protein kinase C triggers the increase in certain receptor density in membranes, thereby enhancing the receptor-coupled cAMP generating responses. The physiological significance of such cross-talk between cellular signaling systems is discussed in comparison with similar previous observations. PMID- 2549058 TI - Structure of the gene encoding the muscle-specific subunit of human phosphoglycerate mutase. AB - We report the isolation and analysis of genomic clones containing the entire gene encoding the muscle-specific subunit of human phosphoglycerate mutase. The gene spans 2.83 kilobase pairs and has a three-exon/two-intron structure that is similar to the organization of the human 2,3-bisphosphoglycerate mutase gene (Joulin, V., Garel, M.-C., LeBoulch, P., Valentin, C., Rosa, R., Rosa, J., and Cohen-Solal, M. (1988) J. Biol. Chem. 263, 15785-15790), in that the second introns of both genes are localized precisely at the same position. A canonical TATA box and an inverted CCAAT box are present immediately upstream of this gene. Comparison with other muscle-specific enzyme genes reveals a conserved 9-base pair element (GGGGCTGGG) in the 5'-flanking region that may be associated with the expression of genes encoding muscle-specific enzymes. PMID- 2549059 TI - Role of phosphatidylinositol in cardiac sarcolemmal membrane sodium-calcium exchange. AB - The purpose of this investigation was to study the effects of a distinct type of phospholipase C on sarcolemmal Na+-Ca2+ exchange. With this phospholipase C (Staphylococcus aureus), treatment of cardiac sarcolemmal vesicles resulted in a specific hydrolysis of membrane phosphatidylinositol. This hydrolysis of phosphatidylinositol also released two proteins (110 and 36 kDa) from the sarcolemmal membrane. Phospholipase C pretreatment of the sarcolemma resulted in an unexpected stimulation of Na+-Ca2+ exchange. The Vmax of Na+-Ca2+ exchange was increased but the Km for Ca2+ was not altered. This stimulation was specific to the Na+-Ca2+ exchange pathway. ATP-dependent Ca2+ uptake was depressed after phospholipase C treatment, but passive membrane permeability to Ca2+ was unaffected. Sarcolemmal Na+,K+-ATPase activity was not altered, whereas passive Ca2+ binding was modestly decreased after phospholipase C pretreatment. The stimulation of Na+-Ca2+ exchange after phosphatidylinositol hydrolysis was greater in inside-out vesicles than in a total population of vesicles of mixed orientation. This finding suggests that the cardiac sarcolemmal Na+-Ca2+ exchanger is functionally asymmetrical. The results also suggest that membrane phosphatidylinositol is inhibitory to the Na+-Ca2+ exchanger or, alternatively, this phospholipid may anchor an endogenous inhibitory protein in the sarcolemmal membrane. The observation that a transsarcolemmal Ca2+ flux pathway may be stimulated solely by phosphatidylinositol hydrolysis independently of phosphoinositide metabolic products like inositol triphosphate is novel. PMID- 2549060 TI - Treatment of A431 cells with epidermal growth factor (EGF) induces desensitization of EGF-stimulated phosphatidylinositol turnover. AB - Epidermal growth factor (EGF) stimulates the turnover of phosphoinositides in A431 cells. In cells that were pretreated with EGF for 30 min at 37 degrees C and then washed to remove surface-bound hormone, a 70-100% decrease in the EGF stimulated production of inositol monophosphate, inositol bisphosphate, and inositol triphosphate was noted when the cells were exposed to the agonist a second time. Since only a 15% decrease in receptor number was observed in these pretreated cells, the loss of responsiveness to EGF for the production of inositol phosphates could not be attributed to a down-regulation of the EGF receptors. These data suggest that pretreatment of A431 cells with high concentrations of EGF leads to a desensitization of the EGF receptor. This desensitization of the receptor by EGF is apparent within 10-15 min of the addition of EGF and is maximal by 30 min. The desensitization appears to be homologous in nature since pretreatment of cells with EGF did not diminish their responsiveness to bradykinin; and conversely, pretreatment with bradykinin did not diminish the subsequent responsiveness of the cells to EGF. Desensitization to EGF was observed in cells in which protein kinase C had been down-regulated by prolonged treatment with 12-O-tetradecanoylphorbol-13-acetate, implying that EGF receptor desensitization is independent of protein kinase C. The desensitizing effects of EGF on growth factor-induced phosphatidylinositol turnover could be prevented by pretreatment of the cells with the calmodulin antagonist trifluoperazine, suggesting that calmodulin may be involved in the regulation of EGF receptor sensitivity. PMID- 2549061 TI - Cyclic GMP levels and membrane current during onset, recovery, and light adaptation of the photoresponse of detached frog photoreceptors. AB - We have used a preparation of isolated, intact rod photoreceptors to correlate the effects of flash illumination on the intracellular cyclic GMP content and the membrane current. We find that the recovery of cyclic GMP levels after brief flash illumination requires approximately twice as much time as the recovery of the membrane current. In contrast, the subsecond kinetics of the cyclic GMP response to light are faster than the kinetics of membrane current suppression. Both cyclic GMP and the membrane current show graded responses to a wide range of flash intensities; however, in a low Ca2+-Ringer's solution, dim flashes can trigger a decrease in cyclic GMP concentration with no corresponding decrease in membrane current. These results suggest that either other factors can regulate the membrane current, or that measurements of total cellular cyclic GMP do not accurately reflect dynamic changes in cyclic GMP concentration in the vicinity of the light-regulated channel. Changes in cyclic GMP concentration in the presence of background illumination exhibit adaptational behavior similar to that observed in a light-adapted photoresponse: acceleration in the response kinetics and a decrease in response amplitude. That this result is observed in rods depleted of internal Ca2+ suggests a Ca2+-independent mechanism by which background illumination can accelerate the cyclic GMP response. PMID- 2549062 TI - Thermodynamic and spectroscopic characteristics of the cytochrome bc1 complex. Role of quinone in the behavior of cytochrome b562. AB - Cytochrome b562 does not behave as a single independent thermodynamic component in preparations of purified quinol cytochrome c reductase. This effect is much more pronounced in quinone sufficient preparations; in such preparations, the epr spectrum of the cytochrome is Eh sensitive, with a peak shift from g = 3.42 to 3.48 occurring as the potential is lowered from 100 mV to 0 mV. The peak shift is dependent on the presence of quinone and can be restored to quinone-depleted preparations by supplementation with ubiquinol 2 if phospholipid depletion is not too severe. The results suggest that cytochrome b562 is strongly interacting with the Qc quinone binding site. PMID- 2549063 TI - Hydroxyl free radical is not the main active species in site-specific DNA damage induced by copper (II) ion and hydrogen peroxide. AB - Site-specific DNA damage by Cu(II) plus H2O2 was investigated by a DNA-sequencing technique. Cu(II) plus H2O2 induced strong DNA cleavage even without piperidine treatment. Piperidine-labile sites were induced frequently at thymine and guanine residues and rarely at adenine residue. A Cu(I)-specific chelating agent, bathocuproine, inhibited the DNA damage. Neither ethanol nor mannitol inhibited it. Of alcohols, tertbutyl alcohol, having relatively low reactivity to hydroxyl free radical, inhibited the DNA damage most strongly. Sodium azide and 1,4 diazobicyclo[2.2.2]octane completely inhibited cleavages at residues of the bases other than guanine. Tris inhibited the DNA damage. The enhancing effect of D2O on DNA damage was not observed. ESR studies using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) showed that the hydroxyl radical adduct of DMPO was formed during the reaction of Cu(II) with H2O2, and that the addition of sodium formate produced the CO2- radical adduct of DMPO more efficiently than expected. ESR studies showed that the nitroxide radical was formed from 2,2,6,6-tetramethyl-4 piperidone in the presence of Cu(II) plus H2O2, indicating the formation of singlet oxygen or its equivalent. The effects of scavengers on DNA damage have considerable correlation with the effects of scavengers on the nitroxide radical production and DMPO.OH formation. The results suggest that the main active species causing DNA damage are more likely copper-peroxide complexes, with similar reactivity to singlet oxygen and/or hydroxyl radical rather than hydroxyl free radical. PMID- 2549064 TI - Mutations in the GTP-binding site of GS alpha alter stimulation of adenylyl cyclase. AB - Mutational replacements of specific residues in the GTP-binding pocket of the 21 kDa ras proteins (p21ras) reduce their GTPase activity. To test the possibility that the cognate regions of G protein alpha chains participate in GTP binding and hydrolysis, we compared signaling functions of normal and mutated alpha chains (termed alpha s) of Gs, the stimulatory regulator of adenylyl cyclase. alpha s chains were expressed in an alpha s-deficient S49 mouse lymphoma cell line, cyc-. alpha s in which leucine replaces glutamine 227 (corresponding to glutamine 61 of p21ras) constitutively activates adenylyl cyclase and reduces the kcat for GTP hydrolysis more than 100-fold. There is a smaller reduction in GTPase activity in another mutant in which valine replaces glycine 49 (corresponding to glycine 12 of p21ras). This mutant alpha s is a poor activator of adenylyl cyclase. Moreover, the glycine 49 protein, unlike normal alpha s, is not protected against tryptic cleavage by hydrolysis resistant GTP analogs; this finding suggests impairment of the mutant protein's ability to attain the active (GTP-bound) conformation. We conclude that alpha s residues near glutamine 227 and glycine 49 participate in binding and hydrolysis of GTP, although the GTP binding regions of alpha s and p21ras are not identical. PMID- 2549065 TI - Synthesis in Escherichia coli of GTPase-deficient mutants of Gs alpha. AB - We have reduced the GTPase activity of the alpha subunit of Gs, the guanine nucleotide-binding regulatory protein that stimulates adenylyl cyclase, by introduction of point mutations analogous to those described in p21ras. Mutants G49V and Q227L differ from the wild type protein in the substitution of Val for Gly49 and Leu for Gln227, respectively (analogous to positions 12 and 61 in p21ras). Wild type and mutant proteins were synthesized in Escherichia coli, purified, and characterized. The rate constants for dissociation of GDP from G49V recombinant Gs alpha (rGs alpha) (0.47/min) and Q227L rGs alpha (0.23/min) differ by no more than 2-fold from that observed for the wild type protein (0.5/min). In marked contrast, the rate constants for hydrolysis of GTP by G49V rGs alpha (0.78/min) and Q227L rGs alpha (0.03-0.06/min) are 4-fold and roughly 100-fold slower than that for wild type rGs alpha (3.5/min). These reductions in the rate of hydrolysis of GTP result in significant fractional occupancy of these proteins by GTP in the presence of the nucleotide, 0.37 for G49V rGs alpha and 0.78 for Q227L rGs alpha, compared to 0.05 for wild type rGs alpha. When reconstituted with cyc- (Gs alpha-deficient) S49 cell membranes or purified adenylyl cyclase, both mutant proteins stimulate adenylyl cyclase activity in the presence of GTP to a much greater extent than does wild type Gs alpha; their maximal ability to activate the enzyme is largely unaltered. The fractional ability of a given Gs alpha polypeptide to active adenylyl cyclase in the presence of GTP correlates well with the fractinal occupancy of the protein by the nucleotide. The mutant subunits appear to interact normally with G protein beta gamma subunits, and their ability to activate adenylyl cyclase is enhanced by interaction with beta adrenergic receptors. These results indicate that the structural analogy that has been inferred between the guanine nucleotide-binding domains of G proteins and the p21ras family is at least generally correct. They also provide confirmation of the kinetic model of G protein function and document mutations that permit the expression in vivo of constitutively activated G protein alpha subunits. PMID- 2549066 TI - The epidermal growth factor receptor tyrosine kinase in liver epithelial cells. The effect of ligand-dependent changes in cellular location. AB - Epidermal growth factor (EGF) activates the intrinsic tyrosine-specific protein kinase of its receptor (EGF-R). We studied the effect of EGF-dependent EGF-R internalization on receptor autophosphorylation and on the appearance of tyrosine phosphoproteins in rat liver epithelial cells. Peak receptor autophosphorylation activity (3- to 6-fold over basal) was found in homogenates of EGF-treated cells at times when the majority of receptors (greater than 90%) had been internalized but not yet degraded (15 to 30 min). Stimulated activity persisted for at least 2 h if EGF-R degradation was blocked by methylamine or 18 degrees C incubation. Detection of stimulated autophosphorylation in homogenates of cells treated with EGF in culture required detergent in the assay. Detergent was not necessary to detect stimulated autophosphorylation when EGF was added directly to homogenates of untreated cells. Immunoblots using antibodies against phosphotyrosine (p-Tyr) demonstrated that EGF treatment of intact cells increased the p-Tyr content of at least seven proteins (EGF-R, 115, 100, 75, 66, 57, and 52 kDa) within 5 s. Incubation of intact cells with EGF at 0 degrees C to prevent endocytosis still resulted in tyrosine phosphorylation of these seven proteins. In contrast, several substrates (120, 78, and 38 kDa) showed delayed increases (45-90 s) in tyrosine phosphorylation at 37 degrees C; their phosphorylation was even slower at 18 degrees C and did not occur at 0 degrees C. In cells incubated with EGF at 18 degrees C or in the presence of methylamine, EGF-R p-Tyr in the intact cell was lost by 2 h even though receptor was not degraded and still exhibited enhanced autophosphorylation in the homogenate assay. These findings suggest that tyrosine phosphorylation in response to EGF occurs predominantly during the initial stages of endocytosis and is mediated for the most part by ligand receptor complexes at the cell surface. A subset of phosphorylations may require intracellular movement. PMID- 2549067 TI - Defective post-translational modification of collagen IV in a mutant F9 teratocarcinoma cell line is associated with delayed differentiation and growth arrest in response to retinoic acid. AB - We have selected a mutant F9 teratocarcinoma stem cell line, RA-5-1, which does not exhibit normal differentiation into parietal endoderm in the presence of retinoic acid, dibutyryl cyclic AMP, and theophylline (RACT). In this report, we demonstrate that the RA-5-1 mutant possesses a prolyl-4-hydroxylase enzyme with a higher Km for a synthetic collagen substrate and that this alteration results in a 6-7-fold reduction in the amount of collagen IV in the medium of RACT-treated mutant cells, as compared to wild type F9 cells. In addition, the collagen IV that is secreted by RACT-treated RA-5-1 cells has an abnormally low molecular weight and contains 6-9-fold less 4-hydroxyproline than the collagen IV secreted by RACT-treated wild type F9 cells. A brief ascorbate treatment can increase the hydroxyproline content of the collagen IV secreted by RACT-treated RA-5-1 cells. A large reduction in the amount of laminin in the medium of RACT-treated RA-5-1 mutant cells is also observed. Concomitant with the reduction in collagen IV and laminin polypeptides in the medium, the expression of several other differentiation-specific mRNAs is delayed in the RACT-treated RA-5-1 cells relative to wild type F9 cells. Moreover, the mutant cells do not exhibit the morphology or the complete growth arrest of wild type terminally differentiated parietal endoderm cells in the presence of RACT. These results suggest that a defect in the post-translational modification of collagen IV in the mutant RA-5-1 prevents the complete expression of the differentiation program in response to RACT. These experiments also demonstrate that the expression of certain differentiation-specific genes is compatible with continued proliferation in the mutant line. PMID- 2549068 TI - The Saccharomyces cerevisiae SOC8-1 gene and its relationship to a nucleotide kinase. AB - The yeast SOC8-1 gene was originally identified by partial complementation of cdc8 mutant strains. We have carried out Bal31 deletion analysis of the SOC8-1 gene to define the minimal size which is required for the complementation of the cdc8 mutation. When the SOC8-1 gene is cloned in a multicopy plasmid, it enables temperature-resistant growth in the cdc8 mutant strain, while the SOC8-1 gene in a single copy plasmid does not. Thus, its suppression of the cdc8 mutant is dosage dependent. The high copy number vector carrying the SOC8-1 gene can complement five different cdc8 alleles, indicating that the suppression is not allele specific. Since CDC8 encodes thymidylate kinase, cells bearing a high copy number plasmid containing SOC8-1 gene were tested for the ability to phosphorylate several nucleoside monophosphates, including UMP, GMP and dTMP. Significantly increased phosphorylation activity was observed, suggesting that SOC8-1 encodes a nucleotide kinase. Both restriction enzyme analysis of the SOC8 1 gene and partial purification of the overproduced kinase in SOC8-1 overproducing strains suggest that SOC8-1 may be allelic with URA6. Consistent with these results, both SOC8-1 and URA6 are located on chromosome XI. Thus, one possible suppression mechanism is that SOC8-1 may provide a trans-acting dTMP kinase activity, bypassing the cdc8 gene defect. PMID- 2549069 TI - Purification of acyloxyacyl hydrolase, a leukocyte enzyme that removes secondary acyl chains from bacterial lipopolysaccharides. AB - Leukocytes contain an enzyme that detoxifies bacterial lipopolysaccharides (also called endotoxins) by removing fatty acyl chains that are attached in acyloxy acyl linkage to the glucosamine backbone of lipid A. We describe the purification of an enzyme that carries out this activity, termed acyloxyacyl hydrolysis, from the HL-60 human promyelocyte cell line. The enzyme is a glycoprotein of apparent Mr = 52,000-60,000 that is found in low abundance (less than 0.001% of the cell lysate protein), principally in the granule fraction of the cells. The protein has two disulfide-linked subunits of apparent Mr = 50,000 and 14,000-20,000, each of which contains N-linked oligosaccharides. This is the first lipopolysaccharide degrading enzyme that has been purified from animal cells. PMID- 2549070 TI - Carboxypeptidase H. A regulatory peptide-processing enzyme produced by human hepatoma Hep G2 cells. AB - Human hepatoma (Hep G2) cells have been shown to secrete nanogram quantities of carboxypeptidase N (Grimwood, B. G., Plummer, T. H., Jr., and Tarentino, A. (1988) J. Biol. Chem. 263, 14397-14401). A second carboxypeptidase with an acidic pH optimum (pH 5.5) is also secreted at levels 2-3-fold greater than carboxypeptidase N. This enzyme was partially purified from the conditioned medium and compared with pure bovine pituitary carboxypeptidase H. The two enzymes behaved in a similar fashion in DE52 ion-exchange chromatography and on gel filtration, with the Hep G2 enzyme being slightly larger than the bovine pituitary enzyme (52-54 versus 50-52 kDa). Both enzymes hydrolyzed COOH-terminal basic amino acids from typical synthetic substrates as well as from natural leuenkephalin peptides and were identical based on pH activity profiles, inhibition by EDTA or guanidinoethyl mercaptosuccinic acid, and stimulation by Co2+ ions. Inhibition of enzyme secretion from Hep G2 cells by tunicamycin indicated that the Hep G2 enzyme was glycosylated. This finding was confirmed by a parallel deglycosylation of the Hep G2 and bovine pituitary carboxypeptidase H enzymes with peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F. Immunoblots using mouse antiserum to bovine pituitary carboxypeptidase H revealed that the Hep G2 enzyme was immunocross-reactive with the bovine enzyme but was slightly larger in size (54 versus 52 kDa). Continuous [35S]methionine labeling and purification to near homogeneity using an affinity matrix corroborated the observations that the secreted Hep G2 carboxypeptidase H was slightly larger than bovine pituitary carboxypeptidase H. The Hep G2-secreted enzyme in pulse-chase experiments was initially detected intracellularly after a 15-min pulse as a single protein of about 54 kDa and was present in the 30-min chase medium with no evidence for pre- or postsecretion proteolytic processing. The human adrenergic cell line IMR-32 continuously labeled with [35S]methionine also secreted carboxypeptidase H of the same size as the Hep G2 enzyme. PMID- 2549072 TI - Influence of GABA and ethylenediamine in the guinea-pig duodenum. AB - 1. GABA induced concentration-dependent transient contractions of the guinea-pig duodenum, but only occasionally evoked small relaxatory responses. The GABA induced contractions were blocked by atropine and tetrodotoxin but were not influenced by hexamethonium; during electrically evoked twitch contractions, GABA had a concentration-dependent inhibitory effect. 2. The concentration-response curve for the contractile effect of GABA was shifted to the right in a dose dependent manner by bicuculline and picrotoxin, with a clear reduction of the maximal effect in the presence of picrotoxin. 3. Homotaurine and delta aminovaleric acid but not baclofen mimicked the GABA-induced contractions; the responses induced by these GABAA receptor agonists were antagonized by atropine, tetrodotoxin and bicuculline. Baclofen concentration-dependently inhibited electrically evoked twitch contractions. 4. Ethylenediamine also had a GABA-like effect, and cross-desensitization developed between GABA and ethylenediamine. 5. The ethylenediamine-induced contractions were not antagonized by thiosemicarbazide; they were reduced by 3-mercaptopropionic acid but the GABA induced contractions were reduced to the same extent. 6. It is concluded that GABA induces contraction of the guinea-pig duodenum by excitation of GABAA receptors on postganglionic cholinergic neurones; a GABAB receptor-mediated inhibitory effect can be observed during electrically evoked twitch contractions. Ethylenediamine mimicks the GABAA receptor-mediated effect probably by a direct effect on the GABAA receptors. PMID- 2549071 TI - Transient increase in phosphatidylinositol 3,4-bisphosphate and phosphatidylinositol trisphosphate during activation of human neutrophils. AB - We recently showed that phosphatidylinositol trisphosphate (PIP3) was present in a unique lipid fraction generated in neutrophils during activation. Here, we demonstrate that the band containing this fraction isolated from thin layer chromatography consists primarily of PIP3 and that only small amounts of radiolabeled PIP3 exist prior to activation. In addition, high performance liquid chromatography of deacylated phospholipids from stimulated cells reveals an increase in a fraction eluting ahead of glycerophosphoinositol 4,5-P2. After removal of the glycerol we found that it coeluted with inositol 1,3,4-P3 when resubjected to high performance liquid chromatography. Thus, we have detected a second, novel form of phosphatidylinositol bisphosphate in activated neutrophils, PI-(3,4)P2. The elevation of PIP3 through the formyl peptide receptor is blocked by pretreatment with pertussis toxin, implicating mediation of the increase in PIP3 by a guanosine triphosphate-binding (G) protein. The rise in PIP3 is not secondary to calcium elevation. Buffering the rise in intracellular calcium did not diminish the increase in PIP3. The elevation of PIP3 appears to occur during activation with physiological agonists, its level varying with the degree of activation. Leukotriene B4, which elicits many of the same responses as stimulation of the formyl peptide receptor but with minimal oxidant production, stimulates a much attenuated rise in PIP3. Isoproterenol, which inhibits oxidant production also reduces the rise in PIP3. Hence formation of PI(3,4)P2 and PIP3 (presumed to be PI(3,4,5)P3) correlates closely with the early events of neutrophil activation. PMID- 2549073 TI - Malignant fibrous histiocytoma in association with hip replacement. PMID- 2549074 TI - Transphyseal fracture fixation using biodegradable pins. PMID- 2549075 TI - Serum factors alter the extent of dephosphorylation of ligands endocytosed via the mannose 6-phosphate/insulin-like growth factor II receptor. AB - Mouse L-cells that contain the cation-independent (CI) mannose 6-phosphate (Man 6 P)/insulin-like growth factor (IGF) II receptor endocytose acid hydrolases and deliver these enzymes to lysosomes. The postendocytic loss of the Man 6-P recognition marker from the cell-associated acid hydrolases was assessed by CI Man 6-P receptor affinity chromatography. 125I-labeled acid hydrolases internalized by L-cells grown at high density were delivered to lysosomes but were not dephosphorylated. In contrast, the same 125I-labeled hydrolases internalized by L-cells maintained at low density were delivered to lysosomes and were extensively dephosphorylated. The dephosphorylation at low density required 5 h for completion suggesting that the phosphatase responsible for the dephosphorylation is located within the lysosomal compartment. Transition from the high to low density state was rapid and was not inhibited by cycloheximide. Medium substitution experiments indicated that serum factors were necessary to maintain the L-cells in the dephosphorylation-competent (low density) state, and that serum-free conditions led to a dephosphorylation-incompetent (high density) state. Addition of IGF II to cells in serum-free medium allowed acid hydrolases subsequently introduced by endocytosis to be dephosphorylated. The results indicate that the removal of the Man 6-P recognition marker from endocytosed acid hydrolases is regulated by serum factors in the growth medium, including IGF II. PMID- 2549076 TI - (Na+ + K+)-ATPase and plasma membrane polarity of intestinal epithelial cells: presence of a brush border antigen in the distal large intestine that is immunologically related to beta subunit. AB - The previously produced monoclonal antibody IEC 1/48 against cultured rat intestinal crypt cells (Quaroni, A., and K. J. Isselbacher. 1981. J. Natl. Cancer Inst. 67:1353-1362) was extensively characterized and found to be directed against the beta subunit of (Na+ + K+)-ATPase as assessed by immunological and enzymatic criteria. Under nondenaturing conditions the antibody precipitated the alpha-beta enzyme complex (98,000 and 48,000 Mr). This probe, together with the monoclonal antibody C 62.4 against the alpha subunit (Kashgarian, M., D. Biemesderfer, M. Caplan, and B. Forbush. 1985. Kidney Int. 28:899-913), was used to localize (Na+ + K+)-ATPase in epithelial cells along the rat intestinal tract by immunofluorescence and immunoelectron microscopy. Both antibodies exclusively labeled the basolateral membrane of small intestine and proximal colon epithelial cells. However, in the distal colon, IEC 1/48, but not C 62.4, also labeled the brush border membrane. The cross-reacting beta-subunit-like antigen on the apical cell pole was tightly associated with isolated brush borders but was apparently devoid of (Na+ + K+)-ATPase activity. Subcellular fractionation of colonocytes in conjunction with limited proteolysis and surface radioiodination of intestinal segments suggested that the cross-reacting antigen in the brush border may be very similar to the beta subunit. The results support the notion that in the small intestine and proximal colon the enzyme subunits are exclusively targeted to the basolateral membrane while in the distal colon nonassembled beta subunit or a beta-subunit-like protein is also transported to the apical cell pole. PMID- 2549077 TI - Cellular remodeling of HCO3(-)-secreting cells in rabbit renal collecting duct in response to an acidic environment. AB - The renal cortical collecting duct (CCD) consists of principal and intercalated cells. Two forms of intercalated cells, those cells involved in H+/HCO3- transport, have recently been described. H+-secreting cells are capable of apical endocytosis and have H+ATPase on the apical membrane and a basolateral Cl-/HCO3- exchanger. HCO3(-)-secreting cells bind peanut agglutinin (PNA) to apical membrane receptors and have diffuse or basolateral distribution of H+ATPase; their Cl-/HCO3- exchanger is on the apical membrane. We found that 20 h after acid feeding of rabbits, there was a fourfold increase in number of cells showing apical endocytosis and a numerically similar reduction of cells binding PNA. Incubation of CCDs at pH 7.1 for 3-5 h in vitro led to similar, albeit less pronounced, changes. Evidence to suggest internalization and degradation of the PNA binding sites included a reduction in apical binding of PNA, decrease in pH in the environment of PNA binding, and incorporation of electron-dense PNA into cytoplasmic vesicles. Such remodeling was dependent on protein synthesis. There was also functional evidence for loss of apical Cl-/HCO3- exchange on PNA-labeled cells. Finally, net HCO3- flux converted from secretion to absorption after incubation at low pH. Thus, exposure of CCDs to low pH stimulates the removal/inactivation of apical Cl-/HCO3- exchangers and the internalization of other apical membrane components. Remodeling of PNA-labeled cells may mediate the change in polarity of HCO3- flux observed in response to acid treatment. PMID- 2549079 TI - Congenital anomalies of the upper extremity. AB - This article has reviewed some of the current methods of management of congenital problems in the upper extremity. Not all problems have been included in this review but some of the landmark advances in this field have been considered. The management of congenital anomalies may change significantly in the future. PMID- 2549081 TI - Translocation of diacylglycerol kinase from the cytosol to the membrane in phorbol ester-treated Swiss 3T3 fibroblasts. AB - The tumor-promoting phorbol ester, 12-O-tetradecanoyl-phorbol-13-acetate, causes a rapid, partial redistribution of 1,2-diacylglycerol kinase from the cytosol to the particulate fraction of quiescent Swiss 3T3 fibroblasts. The inactive alpha form of the phorbol ester does not cause any change in diacylglycerol kinase localization, and depletion of protein kinase C by chronic administration of phorbol ester blocks the redistribution. Phorbol ester has no direct effect on membrane-bound diacylglycerol kinase in 3T3 cells. When phorbol ester is added to 3T3 membranes in the presence of ATP, Mg2+, and Ca2+, there is no activation of membrane-bound kinase, indicating that phorbol ester does not activate membrane bound kinase through phosphorylation by protein kinase C. Stimulation of the cells with phorbol ester increases the total mass of diacylglycerol. In protein kinase C-depleted cells, addition of a cell-permeable synthetic diacylglycerol, dioctanoylglycerol, results in a partial redistribution of cytosolic diacylglycerol kinase to the membrane, also suggesting that the translocation of DAG kinase is regulated primarily by substrate concentration. PMID- 2549080 TI - Phospholipase A2-mediated release of arachidonic acid in stimulated guinea pig alveolar macrophages: interaction with lipid mediators and cyclic AMP. AB - The stimulation of cultured guinea pig alveolar macrophages by the chemotactic peptide N-formyl-L-methionyl-L-leucyl-L-phenylalanine, or by the phospholipid inflammatory mediator platelet activating factor (PAF) induced an increase in arachidonic acid release and its cyclooxygenase products. This release, which was mimicked by the association of threshold concentrations of the calcium ionophore A 23187 and of the protein kinase C activator tetradecanoyl phorbol acetate arose mainly from diacyl- and alkyl-acyl-phosphatidylcholine and phosphatidylinositol. Using [1-14C]arachidonic acid-labeled membranes as an endogenous substrate as well as dioleoyl-phosphatidyl [14C]ethanolamine as an exogenous substrate, we showed that phospholipase A2 activity of stimulated macrophages increases upon stimulation. Treatment of macrophages by prostaglandin E2 decreased the arachidonic acid release elicited by the chemotactic peptide and PAF. Furthermore, prostaglandin E2 increased and PAF decreased the cellular content in cyclic AMP. From these results we suggest that an initial stimulation of alveolar macrophages by a bacterial signal initiates the sequential activation of a phospholipase C and of phospholipase A2, leading to the release of PAF and eicosanoids. These mediators may in turn modulate the cell response by increasing or decreasing cyclic AMP, Ca2+, or diacyglycerol macrophage content. PMID- 2549078 TI - Regulation of macrophage growth and antiviral activity by interferon-gamma. AB - Interferons, in addition to their antiviral activity, induce a multiplicity of effects on different cell types. Interferon (IFN)-gamma exerts a unique regulatory effect on cells of the mononuclear phagocyte lineage. To investigate whether the antiviral and antiproliferative effects of IFN-gamma in macrophages can be genetically dissociated, and whether IFN-alpha and IFN-gamma use the same cellular signals and/or effector mechanisms to achieve their biologic effects, we have derived a series of somatic cell genetic variants resistant to the antiproliferative and/or antiviral activities of IFN-gamma. Two different classes of variants were found: those resistant to the antiproliferative and antiviral effects of IFN-gamma against vesicular stomatitis virus (VSV) and those resistant to the antiproliferative effect, but protected against VSV and encephalomyocarditis virus (EMCV) lysis by IFN-gamma. In addition, a third class of mutants was obtained that was susceptible to the growth inhibitory activity, but resistant to the antiviral activity of IFN-gamma. Analysis of these mutants has provided several insights regarding the regulatory mechanisms of IFN-gamma and IFN-alpha on the murine macrophage cell lines. The antiproliferative activity of IFN-gamma on these cells, in contrast to that of IFN-alpha, is mediated by a cAMP-independent pathway. The antiproliferative and antiviral activities of IFN gamma were genetically dissociated. Variants were obtained that are growth resistant but antivirally protected, or are growth inhibited but not antivirally protected against VSV or EMCV. The genetic analysis indicated that IFN-alpha and IFN-gamma regulate the induction of the dsRNA-dependent P1/eIF-2 alpha protein kinase and 2',5'-oligoadenylate synthetase enzymatic activities via different pathways. Finally, a unique macrophage mutant was obtained that was protected by IFN-gamma against infection by VSV, but not EMCV, suggesting that antiviral mechanisms involved in protection against these different types of RNA viruses must be distinct at some level. PMID- 2549083 TI - How steroid hormones work. AB - Understanding how steroids work has led to improved comprehension of such derangements of hormonal regulation as testicular feminization. This knowledge has found practical application in the treatment of hormone-dependent cancers. Gene therapy, albeit still too primitive to correct steroid-response defects, promises to be a future mainstay of management. PMID- 2549082 TI - The second-messenger system for peptide hormones. AB - There are three major hormone classes--peptide, steroid, and the newly defined growth factors--each with its own system for signal transduction in the cell. Two interdependent theses are proposed for the peptide hormone: that incoming signal transduction requires coupling to a G protein in a second-messenger pathway, and that second-messenger redundancy assures checks and balances in cell regulation. PMID- 2549084 TI - Purification of human muscle phosphoglycerate mutase by fast protein liquid chromatography based on hydrophobic interactions. PMID- 2549085 TI - The potential usefulness of interleukin-2 activated bone marrow cells as an active therapeutic tool against cytomegalovirus infection in a bone marrow transplantation setting. AB - Bone marrow transplantation (BMT) has been used in recent years for the treatment of immunodeficiency diseases, aplastic anemia, and leukemia. However, there are a number of serious problems and limitations associated with autologous or allogeneic BMT. One of these is an increase in opportunistic infections, of which cytomegalovirus (CMV) infection is one of the most important. Cytomegalovirus has been associated with more frequent deaths than any other single agent, with no reproducibly successful or therapy currently available. Recently usage of interleukin-2 or immunomodulation has been suggested as a powerful modality to combat infectious disease. In this study we showed that bone marrow activated in interleukin-2 for 2 days has the ability to lyse spleen cells infected for 3 days with murine CMV (acute infection model) or salivary gland cells infected for 7 days (chronic infection model), while nonactivated bone marrow or natural killer (NK) cells showed no such lysis. The majority of activated cells involved in lysis were antiasialo GM1-, Thy-1+/-, indicating a population of cells other than the natural killer-cell population involved. PMID- 2549087 TI - Multisite evaluation of a monoclonal antibody reagent (Syva) for rapid diagnosis of cytomegalovirus in the shell vial assay. AB - A pre-cytopathic effect (CPE) monoclonal antibody reagent (Syva Co., Palo Alto, Calif.) was evaluated in four laboratories for the rapid detection of cytomegalovirus (CMV) in shell vial cell cultures at 16 to 24 h and 40 to 48 h postinoculation. Results were compared with those obtained by inoculation of the specimen into conventional tube cell cultures that were examined for the presence of typical CMV CPE and subsequently tested by reaction with the monoclonal antibody reagent in an indirect immunofluorescence test. Of 937 specimens, CMV was positive in 184 (20%). CMV was detected twice as frequently in shell vials only (n = 29) as in conventional tube cell cultures (n = 14). Pre-CPE shell vial assay was 91% sensitive (range, 84 to 98%) and 96% specific (range, 93 to 98%) compared with the detection of CPE in conventional tube cell cultures. Overall, 137 of 166 (83%) and 143 of 166 (86%) of the CMV strains were detected at 16 to 24 h and 40 to 48 h postinoculation, respectively. The Syva reagent produced sensitive and specific results for the rapid detection of CMV infection in shell vial cell cultures and reliably confirmed the presence of the virus as detected by CPE in conventional tube cell cultures. PMID- 2549086 TI - A phase I trial of recombinant human interferon-gamma in patients with Kaposi's sarcoma and the acquired immunodeficiency syndrome (AIDS). AB - A Phase I study of recombinant interferon-gamma (rIFN-gamma) was conducted to determine the toxicity and pharmacokinetics of this lymphokine in acquired immunodeficiency syndrome (AIDS) patients with Kaposi's sarcoma (KS). Sixteen patients with AIDS/KS were entered into a fixed-dose trial at either 0.001, 0.01, 0.1, or 1.0 mg/m2 of rIFN-gamma. rIFN-gamma was initially administered either as a single 24-hr continuous iv infusion or as a single im injection, followed 4 days later by a 10-day course of daily therapy by the same route. Following a 1 week washout period, this sequence of administration was then repeated, with the drug given by the alternate route. Pharmacokinetic analysis of the 1.0-mg/m2 group revealed that peak serum levels of up to 153 U/ml occurred 2-4 hr after im injection and that steady-state levels of up to 40 U/ml were reached approximately 7-12 hr after beginning iv infusion. Dose-related toxicities in this trial included fever, headache, fatigue, nausea, and hepatitis, all of which were most severe at the two highest doses. Dose-dependent depression of the total white blood-cell (WBC) count, affecting both granulocytes and lymphocytes, was the most common laboratory abnormality. Natural killer (NK)-cell activity was slightly enhanced at a dose of 0.1 mg/m2 but suppressed at 1.0 mg/m2 of drug; monocyte-mediated cytotoxicity, in contrast, was significantly increased only at the highest dose. No dose-related changes were noted in KS lesions, HLA-DR expression by peripheral blood mononuclear cells, lymphocyte blastogenesis, or the ability to culture cytomegalovirus (CMV) from body fluids. We conclude that a maximally tolerated dose (MTD) for this drug is in the range of 0.1-1.0 mg/m2 and that at least modest evidence of systemic immunomodulation may be seen when rIFN gamma is given at doses at or near this MTD. PMID- 2549089 TI - Comparison of in situ hybridization and monoclonal antibodies for early detection of cytomegalovirus in cell culture. AB - The abilities of each of four diagnostic tests--direct fluorescent monoclonal antibody (direct FA) staining, indirect fluorescent monoclonal antibody (indirect FA) staining, in situ hybridization with biotinylated DNA probes, and in situ hybridization with DNA probes directly linked to enzymatically active horseradish peroxidase-to detect cytomegalovirus soon after culture were compared. Only the indirect FA test and the in situ hybridization method with directly linked HRP DNA probes provided consistent and reliable cytomegalovirus detection as early as 15 h postinfection. PMID- 2549088 TI - Four-year study of rotavirus electropherotypes from cases of infantile diarrhea in Rome. AB - Rotavirus infections were detected in 210 of 675 children with acute diarrhea admitted to a major pediatric hospital in Rome from January 1982 through December 1985. Most of the patients with rotavirus infections were admitted during the winter season in both 1982 and 1985, whereas during the two intermediate years, cases occurred in all months. Among 84 rotavirus samples examined, 14 different electropherotypes were recognized, 2 of which largely predominated over the others. The two electropherotypes were particularly frequent in the 2 epidemic years, altogether accounting for 70.2% of the samples typed, and circulated in distinct periods. None of the viruses showed a short pattern of electrophoretic migration of the genome, indicating a minor involvement of subgroup I rotaviruses in hospitalization-requiring diarrheas occurring in the area surveyed. PMID- 2549090 TI - Double-label immunofluorescence method for simultaneous detection of adenovirus and herpes simplex virus from the eye. AB - The development and application of a double-label immunofluorescence method which has the potential to screen for single or dual infections from any site, in single shell vial cultures, is described. In this study, a total of 1,141 ocular specimens were inoculated in shell vials, centrifuged at 15,000 X g for 1 h, incubated at 37 degrees C for 48 h, and fixed in methanol at room temperature for 15 min. The virus inclusions were detected by staining with a double-label indirect immunofluorescence procedure using mixtures of appropriate first antibodies, followed by fluorescein- and rhodamine-conjugated second antibodies. Each specimen was also inoculated in parallel by the conventional virus isolation method. The sensitivity and specificity of the double-label shell vial procedure were comparable to those with the conventional method, and the former test took only 48 h to complete. The test offers a rapid and simple single-vial procedure which allows for individual or simultaneous detection of multiple pathogens. It results in savings in time and cost over the conventional virus isolation method and other shell vial procedures. PMID- 2549091 TI - Characteristics of different solid-phase immunoassay formats for the measurement of BK virus immunoglobulin M in sera of patients on renal dialysis or with kidney allografts. AB - Solid-phase immunoglobulin M (IgM) antigen capture enzyme immunoassay (AgCEIA) and antibody capture enzyme immunoassay (AbCEIA) were developed for the diagnosis of BK virus (BKV) infections. Of 37 serum samples from renal allograft recipients, 15 were positive for BKV IgM antibody by either AgCEIA, AbCEIA, or antigen capture radioimmunoassay. False-positive IgM results were observed in the AgCEIA in the presence of high levels of BKV IgG antibody (titers greater than or equal to 1:51,200), when rheumatoid factor (RF) titers were greater than or equal to 1:20, or in the presence of high levels of RF (titers greater than or equal to 1:10,240) when BKV hemagglutination inhibition titers exceeded 1:40. False positives due to RF could be eliminated by treatment of sera with anti-human IgG antisera or IgG-coated latex particles. The presence of RF did not, however, produce false-positive results in the AbCEIA. Both AgCEIA and AbCEIA were specific for BKV IgM antibody, as 14 serum samples containing either JC papovavirus, cytomegalovirus, rubella virus, hepatitis A virus, or hepatitis B virus core IgM antibody were negative in both EIAs. Comparison of results obtained for 37 serum samples revealed 14 positive by radioimmunoassay and 11 positive by both AgCEIA and AbCEIA. Both EIAs detected BKV IgM antibody in sera of renal allograft patients and patients on renal dialysis who had reactivated BKV infections persisting for several months after transplantation. PMID- 2549092 TI - Enhanced infectivity of bluetongue virus in cell culture by centrifugation. AB - The effects of centrifugation of the infection of cell culture with bluetongue virus (BTV) were investigated. Baby hamster kidney cells were infected with BTV with or without centrifugation. Viral antigen was detected by immunofluorescence at 24 h in both centrifuged and noncentrifuged cultures. However, after 24 h of infection, the production of PFU in centrifuged cell cultures was 10- to 20-fold greater than that seen in cultures not centrifuged. In addition, centrifugation enhanced the direct detection of PFU from blood samples collected from a sheep experimentally infected with BTV. PMID- 2549094 TI - Detection of herpes simplex virus by using A549 cells in centrifugation culture with a rapid membrane enzyme immunoassay. AB - The RAMP herpes simplex virus (HSV) culture confirmation test was compared with immunofluorescence (IF) staining with a specific HSV monoclonal antibody reagent for the detection of HSV in centrifugation culture. The RAMP test detected 47 of 57 IF-positive specimens (sensitivity, 88.6%) and agreed with 217 of 220 IF negative specimens (specificity, 98.6%). The RAMP test can be performed in less than 15 min and gives an immediate visual result. However, the sensitivity and the false-positive and false-negative results need further investigation. PMID- 2549095 TI - Human papillomavirus detection in urine samples from male patients by the polymerase chain reaction. AB - Human papillomavirus (HPV) detection was performed using the polymerase chain reaction technique on urine samples from 17 male patients with condylomata acuminata in the meatus urethrae. Urine samples from 14 male laboratory volunteers were analyzed as controls. The DNA was extracted and purified from urine sediments, centrifuged at 1,800 and 100,000 x g, and subjected to 40 cycles of amplification with HPV 6 and HPV 11 type-specific anticontamination primers and the heat-stable Taq DNA polymerase. HPV was detected in the urine of 15 (88%) patients. In all positive patients the urine sediments of both the 1,800 and 100,000 x g centrifugation steps contained HPV DNA. Eight patients were found to be positive for HPV 6 DNA, six were positive for HPV 11 DNA, and one was positive for both HPV 6 and HPV 11 DNA. None of the males in the control group was positive for either HPV 6 or HPV 11 DNA. The results demonstrate that HPV can be transported by the urine, probably in exfoliated HPV-infected cells. A similar mechanism may occur during ejaculation, allowing sexual transmission of HPV viruses harbored in the cells of the male genital tract. PMID- 2549093 TI - Characterization by enzyme-linked immunosorbent assay using subgroup- and serotype-specific monoclonal antibodies of human rotavirus obtained from diarrheic patients in Bangladesh. AB - By enzyme-linked immunosorbent assay with group A-, subgroup-, and serotype specific monoclonal antibodies (MAbs), we tested 414 stool specimens collected from pediatric and adult patients hospitalized with acute gastroenteritis between January and June 1988. Of 414 specimens tested, 124 (30%) were positive for group A rotavirus. The subgroup was determined in 110 specimens (88.7%); 16.1% were subgroup I, and 72.6% were subgroup II. Two specimens reacted with both subgroup I- and subgroup II-specific MAbs. Serotype determinations showed that serotype 1 (38.4%) was predominant over serotypes 2 (28.2%), 3 (2.5%), and 4 (23%). Three specimens reacted with more than one serotype-specific MAb. While the frequency of serotype 1 was highest in the two hospitals in Mymensingh, serotype 2 was most prevalent in one hospital in Dhaka. All human rotavirus strains with subgroup I and serotype 2 specificities showed a short electropherotype, and all but one strain with subgroup II and serotype 1, 3, or 4 specificities exhibited a long electropherotype. PMID- 2549096 TI - Isolation of peplomer glycoprotein E2 of transmissible gastroenteritis virus and application in enzyme-linked immunosorbent assay. AB - An enzyme-linked immunosorbent assay (ELISA) based on peplomer glycoprotein E2 was developed for the detection of antibodies to transmissible gastroenteritis virus (TGEV). Purified preparations of E2 were isolated by solubilization of the viral membrane with nonion detergent Nonidet P-40, followed by sucrose density gradient sedimentation. ELISA optical density values with E2 antigen significantly exceeded the indices when other TGEV protein or intact virion antigen was used. It was shown that a virus protein concentration in the E2 preparation of 500 ng per well is sufficient to sensitize the solid phase of microplates. A comparison of the ELISA and the virus neutralization test for the detection of TGEV antibodies was conducted. A significant correlation between the ELISA and the virus neutralization test was shown (r = 0.97). This serological test may be successfully used for various immunologic investigations. PMID- 2549098 TI - Seroepidemiology of Campylobacter pylori infection in various populations. AB - Campylobacter pylori infection has been recognized as being strongly associated with chronic gastritis and duodenal ulceration, but the prevalence of C. pylori infection in a normal population is not known. A serological survey was conducted in four countries with different geographical and socioeconomic status, in a randomly chosen population as representative as possible, by using an enzyme linked immunosorbent assay (ELISA) with a sonic extract of two strains as the antigen. The test had a specificity of 94% when 600 ELISA units was used as the threshold. In France, few children were infected before the age of 10 years. The prevalence then increased gradually to 36.7% in the sixth decade of life. This increasing prevalence of infection with age was also observed in Algeria, Vietnam, and the Ivory Coast but at a higher rate (80 to 90%). In Vietnam, as in France, few children were infected, whereas in Africa, C. pylori infection occurred earlier. The prevalence of infection did not differ with sex for a particular age group; it also did not differ with respect to gastric symptoms, smoking and drinking habits, or urban or rural residence when these potential risk factors were studied. The epidemiological data available on peptic ulcer disease in developing countries compared with developed countries led to the speculation that infection with C. pylori is not a sufficient condition to develop this disease. PMID- 2549097 TI - Enzymatic amplification of human cytomegalovirus sequences by polymerase chain reaction. AB - Polymerase chain reaction (PCR) amplification was used to detect human cytomegalovirus (HCMV) sequences. The fragments selected for amplification were fragments of 130 and 152 base pairs (bp) located at two opposite ends of HCMV strain AD169 EcoRI fragment D. Amplification of the 152-bp DNA was consistently greater than that of the 130-bp DNA. At the optimal Mg2+ concentration of 5 mM, specific PCR amplification of 152-bp DNA with Taq polymerase was sensitive; only one AD169-infected fibroblast cell or 0.01 pg of AD169 fragment D DNA was needed for detection. This specific amplification was also found with various clinical HCMV isolates and peripheral blood cells and urine from patients. In 37 urine samples analyzed simultaneously by PCR and by virus cultivation, identical results were found in 35 samples, while 2 scored positive only by PCR. This suggests that specific amplification of 152-bp DNA is sensitive and can be used for rapid detection of HCMV infections. PMID- 2549099 TI - Modification of shell vial centrifugation method for detection of herpes simplex virus. AB - The shell vial centrifugation method for the detection of herpes simplex virus was modified with the addition of maintenance medium before specimen inoculation rather than after. Sensitivity and specificity values were comparable to those obtained with postcentrifugation medium addition. PMID- 2549100 TI - Evaluation of the Du Pont HERPCHEK herpes simplex virus antigen test with clinical specimens. AB - The HERPCHEK herpes simplex virus (HSV) antigen test (Du Pont, Billerica, Mass.) is a biotinstreptavidin-amplified enzyme-linked immunoassay for the detection of HSV antigen directly in clinical specimens. In a total of 200 mucocutaneous specimens, predominantly lesions from nongenital sites, HERPCHEK had a sensitivity of 90% and a specificity of 99% compared with the shell vial tissue culture method which uses primary rabbit kidney cells. Five false-negative results by HERPCHEK were found with specimens that contained a low percentage of positive cells by the direct immunoperoxidase stain used in a tissue culture method (1 to 9%). The absorbance values of the clinical specimens by the HERPCHEK test do not correlate well with the percentage of positive cells. Our results indicate that the HERPCHEK HSV antigen test, when performed with specimens from lesions, is a sensitive and specific test as compared with shell vial culture. PMID- 2549101 TI - Detection of cytomegalovirus in peripheral leukocytes by different methods. AB - Screening leukocytes for cytomegalovirus (CMV) by shell vial assay gave unsatisfactory results. Of 10 positive specimens (114 samples), only 2 showing CMV could be detected. Disruption of leukocytes prior to their use in shell vial assays increased the sensitivity of CMV detection considerably: of 32 leukocyte specimens from transplant patients with signs of CMV disease, 13 were positive with disruption and only 3 were positive without. With this modification, 17 transplant patients with suspected CMV infection were regularly screened. Viremia could be detected in 10 cases by the shell vial assay and in 11 cases by the direct detection of immediate early antigen. On the average, viremia was detected 11 days before immunoglobulin M or typical clinical symptoms. PMID- 2549102 TI - In vitro amplification of hepatitis B virus sequences from liver tumour DNA and from paraffin wax embedded tissues using the polymerase chain reaction. AB - A 185 base pair fragment from the core-polymerase overlap region of the hepatitis B virus (HBV) genome was amplified using the polymerase chain reaction (PCR). The results were compared with those of Southern blotting on extracted DNA from eight hepatocellular carcinomata. The data agreed with those of Southern blotting in six cases (two positive, four negative) but in two other positive cases PCR failed to amplify HBV sequences. This suggests deletion or mutation, or both, of this viral region in these cases. PCR was also used to amplify HBV sequences from formalin fixed, paraffin wax embedded tissue. Tissue inhibition of PCR occurred which increased with the number of tissue sections. It was present in tissues from different organs and species and fixed by different procedures, thus highlighting the need for a positive control during amplification. Use of formalin fixed Alexander cells, however, showed a sensitivity of one viral copy per 5000 cells. Confirmation of the identity of the PCR products was carried out using PCR-generated biotinylated probes, and suggested the insertion of extra nucleotide sequences or infection with an HBV variant in one case. PMID- 2549103 TI - Osseous defect responses to hydroxylapatite grafting versus open flap debridement. AB - Data from a large number of defects (152) treated with hydroxylapatite (HA) grafts were compared to those from a large number of defects (111) treated by surgical debridement alone (DEBR). Comparison of initial and re-entry surgery measurements showed that both the analysis of relative defect fill and the intrapatient comparisons demonstrated an advantage to the use of HA graft material. 58% of the HA-grafted defects were judged to have a positive (greater than or equal to 50% defect fill) hard tissue response compared to 30% for DEBR. Minimal responses (failures) were 4 times as numerous with DEBR. Similarly, within each patient, HA grafting proved of benefit, particularly regarding hard tissue changes. The use of HA graft materials appears to be of clinical benefit in a majority of defects and a majority of patients. PMID- 2549104 TI - Can disulfiram (Antabuse) be safely co-administered with the monoamine oxidase inhibitor (MAOI) antidepressants? Despite the ability of the major metabolite of disulfiram (diethyldithiocarbamate) to inhibit the enzyme dopamine beta hydroxylase and a report of the induction in rats of central motor disturbances, convulsions, and death following the co-administration of tranylcypromine, there appears to be surprisingly little in the literature that directly addresses this question. PMID- 2549105 TI - Clear cell hidradenoma with whorl formation of squamoid cells: immunohistochemical and electron microscopic studies. AB - Many groups of squamoid cells arranged in whorls were found in a case of clear cell hidradenoma. These cells showed positive staining for carcinoembryonic antigen, but S-100 protein was not detected. Electron microscopic examination showed considerable amounts of tonofilaments, but neither keratohyalin granules nor multivesicular dense bodies were present in these cells. These features suggest that the constituting cells of whorls differentiated toward the eccrine ductal cells, in particular the intraepidermal ductal cells. PMID- 2549106 TI - The comparative clinical cariostatic efficacy of sodium fluoride and sodium monofluorophosphate dentifrices: a review of trials. AB - This report reviews the methods and findings of published clinical studies comparing dentifrices containing sodium fluoride (NaF) and sodium monofluorophosphate (Na2PO3F) for the purpose of determining whether sodium fluoride formulations have superior cariostatic effects. Nine of ten reported trials found a numerical advantage for sodium fluoride dentifrices with highly compatible abrasive systems and the hypothesis that these two fluoride agents have equivalent cariostatic effects is rejected (p = 0.011). Therefore, the collective clinical evidence demonstrates that dentifrices containing NaF have greater cariostatic effectiveness than dentifrices containing Na2PO3F. PMID- 2549107 TI - An in vitro study of the antiviral properties of an alginate impression material impregnated with disinfectant. AB - The antiviral properties of a new alginate impression material impregnated with a disinfectant (didecyldimethyl ammonium chloride) were evaluated in vitro against herpes simplex virus type I and poliovirus type I. The material was exposed to the virus suspension for periods of up to 60 min. The impregnated alginate material showed no clinically acceptable effect against either virus, although log reductions of 1.0 and 1.7 plaque-forming units of herpes virus were obtained after exposure to the material containing the disinfectant for 5 and 60 min, respectively. PMID- 2549108 TI - Porosity determination of cast investment by a wax-infiltration technique. AB - A reproducible method of determining the porosity (pore volume) of cast investment materials is described. This procedure, requiring no highly specialized equipment, involved the infiltration of a test sample with paraffin wax and the weighing of the wax-infiltrated sample in both air and water. Pore volume wax is then extracted with chloroform and the sample reweighed. The difference in weight is used to calculate the porosity of the investment material. Pore-volume data thus obtained are comparable to those resulting from standard techniques and far more sophisticated (and expensive) instrumentation. The porosities of hand-mixed samples of gypsum-bonded, phosphate-bonded and silica-bonded investments were 53.9 per cent, 36.9 per cent and 31.4 per cent and those of the vacuum-mixed samples were 52.5 per cent, 31.3 per cent and 26.9 per cent, respectively. The results show that gypsum-bonded investments were more porous than phosphate-bonded investments while silica-bonded investments were the least porous. Hand-mixed samples appeared to have greater porosity than vacuum mixed samples. PMID- 2549109 TI - Detection of leukotrienes B4, C4 and of their isomers in arterial, mixed venous blood and bronchoalveolar lavage fluid from ARDS patients. AB - Seven patients with the adult respiratory distress syndrome (ARDS) were studied. As a control group we used 6 surgical patients who underwent minor surgical operation (inguinal hernia). For both groups the same sample collection and analysis was used. The presence of leuktorienes (LTs) B4 and C4 and of their isomers 11-trans LTC4 and delta 6-trans-12-epi LTB4 was determined in arterial, mixed venous blood and in bronchoalveolar lavage (BAL) fluid. The samples, analysed by reverse phase high performance liquid chromatography (RP-HPLC), showed a similar chromatographic picture among ARDS patients, while the control group showed no detectable amounts of LTs in BAL or blood. The distribution of these arachidonic acid metabolites in mixed venous blood, arterial blood and BAL seems to suggest pulmonary metabolism and/or inactivation. It is suggested that these mediators act as humoral factors in pathogenesis of the ARDS. PMID- 2549110 TI - Influences on cortisol and noradrenergic turnover of healthy controls and depressed patients during L-tryptophan loading. AB - To investigate the interrelationships between the noradrenergic and serotonergic systems and the hypothalamic-pituitary-adrenal (HPA) axis, we measured the excretion of 3-methoxy-4-hydroxyphenylglycol (MHPG) and free cortisol (UFC) in 24 h urine both before and after loading with 2 g L-tryptophan (L-TRP) orally in 24 healthy controls and 33 depressed patients categorized according to DSM-III. Loading with L-TRP reduced MHPG excretion significantly by 56% in major depressives, whereas in healthy controls and in minor depressives no significant effects were observed. Although basal MHPG did not differ among these study groups, loading with L-TRP resulted in MHPG excretion in major depressives being significantly lower than in healthy controls and minor depressives. Loading with L-TRP increased UFC significantly by 49% only in major depressives with associated features. After L-TRP those patients exhibited significantly increased UFC as compared with all other depressives and controls, whereas basal UFC values did not differ among the study groups. In baseline conditions there were no significant correlations between MHPG and UFC excretion. PMID- 2549111 TI - Lubricating coolants for high-speed dental handpieces. AB - Cutting studies were performed on tooth enamel, dental amalgam, and composite resin through use of carbide burs and diamond stones with three irrigants. A mixture of water, alcohol, and glycerol significantly increased the rate of material removal when enamel was cut with diamonds and when dental amalgam was cut with diamonds and carbide burs. In contrast, the rate of material removal for composite resin was significantly faster with use of carbide burs than diamond stones, and the irrigant improved the cutting action for carbide burs. The studies indicate that chemomechanical effects operate with dental substrates, but the effect varies with the material being cut and the cutting tool used. The chemomechanical effects observed here significantly improve the rate of material removal with diamond stones and carbide burs. PMID- 2549112 TI - Nutrient intake of marathon runners. AB - Three-day food records from a large group of marathon runners (291 men, 56 women) were analyzed and compared to various standards of dietary quality. Intake by the runners exceeded two-thirds of the Recommended Dietary Allowance for all nutrients except vitamin D and zinc in female runners. Energy and nutrient intake levels in the marathon runners were higher than those of the general U.S. population except for total fat, vitamin B-12, and zinc in both men and women and sodium in men. In general, runners consumed better diets than the general population. However, both caloric intake and percent energy as carbohydrate were lower than recommended for individuals engaging in endurance exercise. The need for professional help in planning high-carbohydrate diets for runners is indicated. Increasing amounts of training were not associated with significant increases in nutrient densities across three activity categories within this group of runners. However, more than 75% of the runners perceived their diets to be much improved after they began regular training (men 8.2 +/- 0.3 years of running experience, women 6.7 +/- 0.6). Together, these data suggest that regular moderate-to-heavy endurance exercise is associated with a better quantity and quality of nutrient intake than that found in the general population, which may in part be related to the adoption of such exercise by previously sedentary individuals. PMID- 2549114 TI - Mineralocorticoid effector mechanism of liquorice derivatives in human mononuclear leukocytes. AB - The mineralocorticoid effector mechanism of glycyrrhetinic acid and of its ester derivative carbenoxolone was evaluated in human mononuclear leukocytes by radioreceptorassay and measurement of intracellular sodium and potassium after incubation of the cells with or without the drugs. The affinity of both compounds for mineralocorticoid receptors in this human model was also compared with that found in kidney cytosol from adrenalectomized rats. Glycyrrhetinic acid possesses a measurable affinity for mineralocorticoid receptors in mononuclear leukocytes, which is 1:3,000 that of aldosterone itself. Carbenoxolone does not bind to the receptors in mononuclear leukocytes, in contrast with kidney cytosol where the drugs show a parallel affinity. The mean intracellular content of sodium in mononuclear leukocytes from 7 volunteers was 35% higher (p less than 0.05) after incubation with 3 mumol glycyrrhetinic acid than after incubation with an equivalent amount of carbenoxolone, or in medium alone. The effect of glycyrrhetinic acid was completely reversed by addition of canrenone for the period of incubation. We conclude that the syndrome of pseudohyperaldosteronism from carbenoxolone is thus probably not related to a direct agonist effect of the drug at the level of mineralocorticoid receptors, but that any action must follow in vivo conversion into glycyrrhetinic acid by hydrolysis. PMID- 2549113 TI - Effects of the antidopaminergic drug veralipride on LH and PRL secretion in postmenopausal women. AB - Patterns of LH and PRL secretion have been evaluated in 15 postmenopausal women before and after the chronic blockage of the D2 dopamine receptors with veralipride (100 mg twice daily, for 30 days). In addition, the possible influence of the antidopaminergic drug on the activity of the endogenous opioid system has been evaluated by the infusion of the opioid antagonist naloxone, performed before and during veralipride administration. Mean plasma LH levels were significantly blunted (p less than 0.05) and mean plasma PRL levels were significantly increased (p less than 0.001) by veralipride administration. The frequency of both LH and PRL secretory pulses was not modified, while the mean pulse amplitude of both hormones was significantly increased (p less than 0.05 for LH; p less than 0.001 for PRL) by veralipride administration. In untreated postmenopausal women naloxone infusion did not modify LH secretion. Following veralipride, the infusion of naloxone significantly increased (p less than 0.05) the mean plasma LH levels, had no influence on the frequency and significantly reduced (p less than 0.01) the amplitude of LH pulses, expressed as the percent increase from the nadir to the peak. Both before and after veralipride administration, naloxone failed to modify the pattern of PRL secretion. In untreated postmenopausal women, the percentage of concomitant PRL and LH pulses was significantly higher (p less than 0.001) during naloxone than during saline infusion, and this effect was amplified by veralipride administration (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549115 TI - Pancreatic insuloma causing Cushing's syndrome. AB - The advent of new diagnostic procedures such as petrosal venous sampling and CRF testing has improved the possibilities of precise location of the site of hormonal overproduction in Cushing's syndrome. We report on a case of ectopic ACTH-dependent Cushing's syndrome in which the use of such techniques helped to locate the tumor. A 45-year-old woman with definite ACTH-dependent Cushing's syndrome was referred to our clinic for evaluation. ACTH, cortisol and urinary 17OHCS failed to suppress on high dose dexamethasone oral administration. Computerized tomography showed bilateral adrenal hyperplasia and an empty sella turcica. ACTH and cortisol did not increase after CRF administration. Staged caval system catheterism via both femoral veins up to the inferior petrosal sinuses, including after CRF administration, demonstrated no ACTH gradient between staged samples and simultaneous peripheral venous blood. This suggested a tumor drained by the portal system. An abdominal ultrasonography demonstrated a pancreatic mass, that was surgically excised and appeared as a benign islet cell tumor. Portal vein ACTH promptly decreased after tumor excision and postoperative peripheral cortisol and ACTH were normal. On immunostaining the tumor cells proved to be positive for ACTH. PMID- 2549116 TI - Angiotensin II and glucocorticoid release: direct effect at the adrenal level and modulation of the adrenocorticotropin-induced glucocorticoid release. AB - Angiotensin II (A II) stimulates adrenal glomerulosa cells releasing mineralocorticoids; however, little is known about the A II effect on glucocorticoids output. The present study has been designed in order to see if A II could modify in vitro spontaneous and ACTH-induced corticosterone (B) release from both fasciculata-reticularis enriched and total adrenal cells. The results indicate that A II at 10(-12), 10(-11), 10(-10) and 10(-6) M concentrations did not modify basal B production and A II 10(-9), 10(-8) and 10(-7) M decreased basal B production from total adrenal cells. Whereas A II (10(-10)-10(-6) M) stimulated B release from fasciculata-reticularis enriched cells. On the one hand, 10(-8) M A II significantly decreased ACTH-elicited B release from total adrenal cells; effect completely abolished by saralasin (SAR, 10(-8) M), a specific A II receptor blocker. On the other hand, 10(-8) M A II did not modify ACTH-induced B release from fasciculata-reticularis enriched cells. Finally, 10( 10) to 10(-6) M A II and 22 pM ACTH stimulated aldosterone output from total adrenal cells; while, fasciculata-reticularis enriched cells did not secrete any measurable amount of aldosterone under basal condition and after incubation with A II. These data further suggest a regulatory role of A II in the release of glucocorticoids from the adrenal gland. PMID- 2549117 TI - Influence of nutritional state on the disposal of orally and intramuscularly administered iodized oil to iodine repleted older children and adult women. AB - Iodinated oil (Ethiodol, 1 or 2 ml) was administered po or by im administration to adult women and older children in rural highland Ecuador who were either well nourished or malnourished to determine the effect of nutritional status on the disposal rate of iodine. These subjects resides in a region previously severely deficient in iodine, but this had been corrected in these subjects by prior administration of iodinated oil or by use of iodized salt or both. Malnutrition as determined by the conventional standards of height for age was associated with a significantly shortened retention time of the administered iodine, whether given po or im. The half life of retention was approximately half in the malourished of that in the well nourished. If these findings can be extrapolated to chronically iodine deficient subjects, then malnourished populations in need of iodine supplementation should either receive higher dosages than those conventionally employed or more frequent dosage. PMID- 2549118 TI - Isolated adrenocorticotropin deficiency and empty sella associated with hypothyroidism. PMID- 2549119 TI - Effect of age on acute regulation of beta-adrenergic responses in mononuclear leukocytes. AB - We studied the acute regulation of beta-adrenergic receptors and cAMP production in mononuclear leukocytes from young and old human subjects. After one hour of supine rest, healthy young and elderly subjects had similar beta-adrenergic receptor density and cAMP responses to isoproterenol. After 10 min of standing, beta-adrenergic receptor density and cAMP response increased in the young subjects. Elderly subjects had a similar increase in cAMP responses after standing, but no change in beta-adrenergic receptor density. Lymphocyte subsets and percent monocytes were not altered by age or posture, suggesting that this was not an artifact of changes in cell populations. Incubation in vitro of cells from both groups with catecholamines, at concentrations comparable to those achieved in plasma after standing, resulted in enhanced isoproterenol-mediated cAMP responses, but no change in beta-adrenergic receptor density. These data suggest that acute regulation of adrenergic signaling is affected by age, mediated in part by catecholamines, and may be relevant in the study of acute cardiovascular regulation. PMID- 2549120 TI - Taurocholate transport by liver plasma membrane vesicles is not altered in cirrhotic rats. AB - Recently, we have shown that taurocholate transport is impaired in hepatocytes isolated from CCl4 cirrhotic rats. Na+,K+-ATPase activity depends on the lipid composition of the surrounding membrane. Therefore, we performed this study in order to detect differences in plasma membrane composition and membrane functions between livers of CCl4 cirrhotic (n = 17) and of control rats (n = 15). After biochemical characterization of the animals we isolated basolateral and canalicular membrane vesicles and determined membrane enzyme activities, transport functions and lipid composition. We found no differences in the isolation characteristics of the plasma membranes between the two groups. The lipid composition of the membrane fractions was not altered, except for a lower cholesterol content in the canalicular membranes of the cirrhotic group (200 +/- 15 vs. 246 +/- 18 micrograms/mg protein, P less than 0.05). Taurocholate transport into basolateral membrane vesicles and marker enzyme activities of the membrane fractions were also equal in control and cirrhotic animals. We conclude that the plasma membrane composition and membrane enzyme/transport activities have returned to normal in CCl4 cirrhotic rats 14 days after cessation of exposure to CCl4. Thus, a disturbed transport system is not the cause for the observed decreased taurocholate transport into hepatocytes from cirrhotic rats. Even a cirrhotic liver has a high potential for recovery after acute CCl4 intoxication. PMID- 2549121 TI - The application of quantitative cytochemistry to study the acinar distribution of enzymatic activities in human liver biopsy sections. AB - The zonal distribution of enzyme activities was measured by quantitative cytochemistry in cryosections of liver from three normal children and five infants with idiopathic hepatitis of infancy. Optimal conditions for cytochemical reactions were first validated in rat liver and subsequently used in human livers to quantify zonal activities of acid phosphatase (AP), succinate dehydrogenase (SDH), glutamate dehydrogenase (GDH), glucose-6-phosphatase (G6P) and NADPH dehydrogenase (ND). In normal rat and human livers, activities were greater for SDH and G6P in periportal and for GDH and ND in perivenular hepatocytes, while AP was evenly distributed along the sinusoids. In five infants with idiopathic hepatitis of infancy (IHI), a similar trend of distribution was observed for the two mitochondrial (SDH and GDH) and the two microsomal (G6P and ND) enzymes, although the distribution gradient was less pronounced than, in normal livers. AP showed a mildly greater periportal than perivenular activity. This preliminary study shows that a similar metabolic zonation exists for these enzymes in human livers as is observed in rats. PMID- 2549122 TI - Quantitative immunocytochemical localization of Na+,K+-ATPase in rat ciliary epithelial cells. AB - Ultrastructural localization of Na+,K+-ATPase in rat ciliary epithelium was investigated quantitatively by the protein A-gold technique, using an affinity purified antibody against the alpha-subunit of Na+,K+-ATPase. Immunoblot analysis showed that the antibody bound specifically to the alpha-subunit of Na+,K+-ATPase in the ciliary body. Gold particles were found mainly on the basolateral surfaces of both the pigmented epithelial (PE) and nonpigmented epithelial (NPE) cells with an approximately twofold higher labeling density in the PE cells. A few gold particles were also found on the apical and ciliary channel surfaces of the PE cells, whereas no significant binding was found on the apical surfaces of the NPE cells. The basolateral surfaces of PE and NPE cells are markedly infolded and are much greater in area than the apical surfaces. This means that Na+,K+-ATPase is almost exclusively located on the basolateral surfaces of both the NPE and PE cells. We suggest that the Na+,K+-ATPase of both the NPE and PE cells play an important role in the formation of aqueous humor. PMID- 2549124 TI - Effects of the tumor promoter, phorbol 12-myristate, 13-acetate, on the epidermal adenylate cyclase system: evidence for adenylate cyclase-regulation by protein kinase C. AB - Exposure of pig epidermal sheets to the protein kinase C (PKC) activator, phorbol 12-myristate, 13-acetate (PMA) resulted in an increase in forskolin-induced cyclic AMP accumulation in the epidermis. Cholera toxin-induced cyclic AMP accumulation was moderately increased by PMA treatment, but this was not statistically significant. On the other hand, receptor-mediated adenylate cyclase responses (beta-adrenergic-, prostaglandin E-, adenosine-, and histamine (H2) adenylate cyclase responses) were significantly decreased. These PMA-induced effects on the epidermal adenylate cyclase system were mimicked by 1-oleoyl-2 acetyl-glycerol, a membrane-permeable synthetic diacylglycerol, and by the non phorbol PKC activator, mezerein. 4-O-methyl PMA, a very weak PKC activator, had no effect on adenylate cyclase responses of the epidermis. The addition of the PKC inhibitor, H-7 (1-(5-isoquinoline-sulfonyl)-2-methyl piperazine dihydrochloride), to the incubation medium significantly inhibited the effect of PMA on forskolin-induced cyclic AMP accumulation. Furthermore, following H-7 treatment, the epidermal receptor-adenylate cyclase responses were significantly increased. These results indicate that PKC modulates epidermal adenylate cyclase responses resulting in an increase in forskolin-induced cyclic AMP accumulation and a decrease in receptor-adenylate cyclase responses of the epidermis. PMID- 2549123 TI - A functional study of the development of the cardiac sympathetic neuroeffector junction in the SHR. AB - We studied the function of the cardiac sympathetic nerve varicosity in isolated right atrial preparations of spontaneously hypertensive rats (SHR) and Wistar Kyoto (WKY) rats at 4, 9, 14, 20 and 50 weeks of age. Cumulative concentration atrial period response (C-R) curves to isoprenaline showed similar maximum response and sensitivity (EC50) at all ages but there was an age-related fall in resting atrial rate. Similar results were found for methoxamine (alpha 1 adrenoceptor agonist) although the maximum response was significantly less than for isoprenaline. The time-dependent recovery (T 1/2) of the fall in atrial period in response to sympathetic nerve stimulation from electrical field pulses (1-32 at 1 Hz) was enhanced by neuronal uptake inhibition by desipramine (0.1-1 mumol/l), to a similar degree at all ages. Pre-junctional alpha 2-adrenoceptor stimulation by clonidine caused progressively more inhibition of the number of field pulses-fall in period relationship with age. SHR atria were similar to WKY rat atria at all ages except for a further impairment of the development of pre junctional alpha 2-adrenoceptors. These studies indicate that the function of the cardiac sympathetic varicosity matures early (by 4 weeks) and overall there is very little impairment in SHR versus WKY rat atria. PMID- 2549125 TI - Treatment of cutaneous leishmaniasis with Pentostam or cryosurgery. AB - Cutaneous leishmaniasis (CL.) caused by L. major or L. tropica is endemic in many countries in the Middle East particularly Saudi Arabia. The disease is characterized by spontaneous healing and sterilizing immunity. However, medical treatment is always indicated particularly those due to L. tropica to reduce the reservoir of infection. In this study parasitologically proven patients with CL. were treated with Pentostam (57) or Cryosurgery (38). Both gave satisfactory results. However, the disadvantages of each line were discussed. There is a need for a drug which is cheaper, less toxic, safe, more effective and more easily administered than those currently available. PMID- 2549126 TI - Correlation of an estrogen receptor-related phosphoprotein with histopathological features in breast cancer. AB - A series of 65 cases of different histological types of breast carcinoma was investigated for the immunohistochemical location of the estrogen receptor related, 29 kD phosphoprotein using the ER-D5 monoclonal antibody. The ER-D5 response is heterogeneous in relation to some therapeutic limitations and is correlated with histopathological features of the tumors and survival. The main parameters for evaluation of breast cancers are reviewed, both those that are statistically correlated and those that are not apparently always correlated but are known to have considerable biological meaning, such as the ER-status of tumors. PMID- 2549127 TI - [A clinical study of six surgically treated patients with malignant tumors arising from chronic pleuritis and pyothorax]. AB - During the clinical course of chronic pleuritis and pyothorax, we frequently have found abnormal clinical signs such as increased abnormal chest shadows, subjective symptoms, such as chest pain and bloody sputum, and the appearance of atypical cells in the pleural effusion. It is very difficult and important to diagnose differentially worsening of chronic inflammation and occurrence of malignant tumors. We have surgically treated six patients with malignant tumors arising from chronic pleuritis and pyothorax, who had the above abnormal signs. All of them were made, and their average age was fifty three years. Two patients exhibited increased abnormal chest shadows, 5 had subjective symptoms (5 with chest pain, 2 with bloody sputum and 2 with fever), and 1 had atypical cells in the pleural effusion. Two patients had suffered from chronic pleuritis, who both had received artificial pneumothorax therapy for pulmonary tuberculosis, and the other 4 patients had suffered from chronic pyothorax, with mean duration of thirty years. The pathological diagnoses were 2 cases each of malignant pleural mesothelioma, squamous cell carcinoma, and malignant fibrous histiocytoma. The two patients with squamous cell carcinoma had pleurocutaneous and bronchopleural fistula. The operative methods were tumor resections with pleuropneumonectomy performed in 2 patients and pneumonectomy in 1. Absolute non-curative operation was performed in 1 patient, and postoperative radiotherapy in 3 patients. All patients died of tumor recurrence, although five of them underwent reoperation, chemotherapy and radiotherapy. Their duration of survival ranged from 3 to 15 months with a mean of 9 months after appearance of malignant tumor signs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549128 TI - [A case of benign fibrous histiocytoma of the lung in a child]. AB - A case of benign fibrous histiocytoma of the lung in a 8 years old boy was presented. He was first admitted in May, 1983, with recurrent pneumonia. Chest X ray showed a ill-defined mass in the right lower lobe. Bronchoscopy revealed a round tumor, 1 cm. in maximum diameter, with complete obstruction of the Truncus Intermedius. Endoscopic resection was performed and partial obstruction of the Truncus Intermedius remained. He was re-admitted with pneumonia of the right middle lobe in March, 1986. Bronchoscopy showed severe stenosis of the Truncus Intermedius. Right middle and lower lobectomy was performed. The 4 X 4 X 3 cm tumor was located in the median of the Truncus Intermedius. Microscopically, the lesion composed of fibroblast-like cells and histiocyte-like cells. The patient's post-operative course was uneventful. He has no signs of local recurrence or metastasis. We believe, this is the first reported primary benign fibrous histiocytoma of the lung in a child in Japan. PMID- 2549129 TI - [Phrenic nerve conduction disorder after open heart surgery--electrophysiological study]. AB - In 32 consecutive adult patients undergoing heart surgery, the induced diaphragmatic muscle action potential was measured. Phrenic nerve conduction disorder was defined as disappearance of muscle action potential (Edi) and conduction time (CT). Phrenic nerve conduction disorder was observed in 10 patients (31%); 8 patients on the left side and 2 patients on both sides. In non conduction disorder group (22 patients), Edi and CT were measured. Edi of the right side decreased significantly from preoperative value of 705 +/- 318 microV to 445 +/- 285 microV at 1-3 days after operation (stage I) and to 559 +/- 314 microV at 7-10 days after operation (stage II) (p less than 0.05). CT of the right side prolonged significantly from 7.1 +/- 0.7 msec before operation to 7.44 +/- 0.97 msec at postoperative stage I and to 7.40 +/- 0.21 msec at postoperative stage II (p less than 0.05). For the left phrenic nerve, Edi showed significant (p less than 0.05) decrease from 803 +/- 338 microV before operation to 429 +/- 251 microV at the postoperative stage I and 620 +/- 350 microV at the postoperative stage II. In the conduction disorder group, incidence of atelectasis, diaphragm elevation and pleural effusion as documented by chest roentgenographic findings were higher than those of non-conduction disorder group (p less than 0.01). Moreover, the lowest temperature of the myocardium during operation was significantly (p less than 0.05) lower for conduction disorder group as compared to non-conduction disorder group. We believed that it is necessary to develop a innovative method for preventing the phrenic nerve from cold injury. PMID- 2549130 TI - Control of therapy with microcirculatory and phlebotropically active drugs in patients with congenital and acquired venous insufficiency. AB - During the treatment of two patients with Klippel-Trenaunay syndrome interesting observations of the efficacy and the side effects of DHE were made. This led to the decision to carry out a validity study in 12 patients suffering from chronic venous insufficiency (CVI). The patients were treated with 0.25 mg or 0.5 DHE intravenously, and after that with 7.5 mg orally for one week. Before and after treatment measurements of venous capacity, microcirculatory parameters and rheological parameters were performed. Following the i.v. injection of 0.25 to 0.5 mg DHE the venous capacity decreased significantly in a dose-dependent way. The flow of erythrocytes in capillaries measured under resting condition was significantly lower and peak flow of reactive hyperemia decreased. No relation was found between the dose of DHE administered and the particular side effects (stomach trouble, increase in diastolic blood pressure) in 2 of the 12 patients. After oral treatment patients showed signs of subjective improvement of their complaints. On the basis of the results, the validity of non-invasive angiological tests is discussed. PMID- 2549131 TI - [A study on maternal humoral immune responses to villous trophoblast using Epstein-Barr virus]. AB - To determine whether or not maternal antibodies are generated during pregnancy, B lymphocytes from puerperal women were transformed using Epstein-Barr virus (EBV). Four weeks after infection with EBV human immunoglobulin (Ig) levels were measured in the culture supernatants by ELISA. In most wells infected B lymphocytes produced a great amount of Ig. The reaction of the culture supernatants from Ig-produced wells with placental tissues of their own was examined by immunohistochemically (by an indirect immunoperoxidase technique). The second layer antibody, goat anti-human Ig peroxidase conjugate was diluted to such a concentration that it did not react to placenta sections without the first layer, that is, culture supernatants. Some of supernatants were found to have IgM which reacted to syncytiotrophoblast. Such positive staining was observed in three of ten multiparous women and in one of two primigravidas. In conclusion, it seems likely that normal pregnant women recognize trophoblasts of their own and make antibodies to them. PMID- 2549132 TI - [Clinicopathological study of malignant ovarian germ cell tumor]. AB - In treating malignant germ cell tumors clinicopathologically at Kurume University Hospital, for 18 years, the total number of patients was 112, including 25 yolk sac tumors (YST), 10 mixed form germ cell tumors (MF), 36 dysgerminomas (DYS), 26 immature teratomas (IT), 2 choriocarcinomas (CHO) and 13 dermoid cysts with malignant transformation (DCMT). The average age of patients in each group was 25.9 years for DCMT patients. When DCMT cases were excluded, ages was closely related to the age of menarche in each patient. The 5 year survival rate for each germ cell malignancy after the first operation was 31.6% for YST cases, 20.0% for MF, 76.7% for DYS, 60.0% for IT and 50.0% for DCMT cases. Despite currently used aggressive kinds of chemotherapy, the clinical stage still had a prognostic significance in all malignant germ cell tumors except dysgerminoma. Since histological maturation of the tumor tissue following treatment courses was observed in YST and IT cases, the possibility of a relationship between prognosis and the histogenetic process was shortly discussed. PMID- 2549133 TI - [Fundamental and clinical studies on human papillomavirus infection in the uterine cervix especially by Vira Pap (dot blot) method]. AB - Female genital organs infected with human papillomavirus (HPV) have drawn attention as STD and in connection with the mechanism of carcinogenesis. Recently, we used a simplified HPV detection kit, Vira Pap method in clinical tests. To estimate the usefulness of the Vira Pap method in clinical application, a comparison was made between the results of the Vira Pap method and those of the conventional method. Furthermore, an investigation was made of the relationship between uterine-cervical lesions and the types of HPV. The following findings were obtained: 1. It was demonstrated that the Vira Pap method was superior to the conventional methods and was almost equal to the Southern blot method. 2. The cases in which HPV infection were confirmed by the Vira Pap method were further analyzed and HPV typed by the Southern blot method. And, types 6, 11, 18, 31, 33, 35 and unclassifiable types were found. 3. HPV 16, 18, and 33, which are said to be closely related to carcinoma, were detected in cases of chronic cervicitis as well. In these cases, further investigation seems to be required. PMID- 2549134 TI - [Carcinoma of male breast--with special reference to noninvasive carcinoma]. AB - Seven cases of carcinoma of male breast were reported. The mean age of them was about 65 years, 17 years older than that of female breast cancer. Six tumors out of 7 were located under the areola. By histological examination, 4 of 7 cases were proved to be noninvasive ductal carcinoma, and the others are invasive ductal carcinoma (2 : scirrhous, 1 : solid-tubular). We focused on clinicopathological features of noninvasive carcinoma. There were two points to be mentioned. One is the nipple discharge as a chief complaint, and the other is cyst formation as a macroscopic observation. These features are characteristic to noninvasive carcinoma and contribute to diagnosis. Therefore, for screening the mass of male breast, ultrasonography (U.S.) is most useful. For preoperative final diagnosis, aspiration or smear cytology is essential. In regard to postoperative survival, all of the 3 invasive cases were dead but all of the 4 noninvasive cases are alive. So the prognosis of noninvasive carcinoma of male breast does not appear to be worse than the female one. These observations indicate that the prognosis of carcinoma of male breast can be improved by early diagnosis and appropriate surgical therapy. PMID- 2549135 TI - [A case of Klippel-Trenaunay-Weber syndrome associated with a short affected leg, spinal AVM and disseminated intravascular coagulation]. PMID- 2549136 TI - [Treatment of disseminated cytomegalovirus infection with ganciclovir in a patient with acquired immunodeficiency syndrome]. PMID- 2549137 TI - [A case of MCTD with pulmonary hypertension showing elevated serum angiotensin I converting enzyme activity]. PMID- 2549138 TI - Evaluation of ileal W pouch-anal anastomosis for restorative proctocolectomy. AB - Clinical defaecatory function, neorectoanal manometry and pouchography were assessed in 16 patients treated by restorative proctocolectomy with ileal W reservoir. The duration after ileostomy closure was 6 to 28 months (mean 17 months). There were no operative deaths and no failures where the reservoir had to be removed. Partial anastomotic dehiscence occurred in one patient, and intestinal obstruction requiring laparotomy in two. Anastomotic stricture, which could be corrected easily by dilatation, occurred in three patients. Daily stool frequency was 4.3 +/- 1.2 at 6 months after ileostomy closure, 3.8 +/- 1.2 at 12 months, and 3.3 +/- 1.0 at 24 months. The clinical score for neorectal function gradually and steadily improved with time as well as daily stool frequency. In the manometric and pouchographic studies, mean anal canal length (3.4 +2- 0.6 cm), mean maximal anal sphincter resting pressure (57.1 +/- 9.7 cm water) and mean maximal reservoir resting pressure (4.3 +/- 2.0 cm water) tended to be less than normal controls but not significantly so. Neorectoanal inhibitory reflex disappeared completely or was greatly decreased in all patients. However, all were capable of spontaneously controlled defaecation. There was an inverse linear relationship between daily stool frequency and maximal tolerated reservoir volume (p less than 0.01). There were inverse linear relationships also between daily stool frequency and horizontal diameter of the reservoir measured on pouchography (p less than 0.05), and daily stool frequency and dilatation ratio of the reservoir (p less than 0.01). From these results, we conclude that a large and wide reservoir allows better defaecatory function. PMID- 2549139 TI - Hepatoblastoma: case report and literature review. AB - Hepatoblastoma is the most common primary malignant hepatic neoplasm in children and must be differentiated from other hepatic tumors such as hepatocellular carcinoma, metastatic neuroblastoma, hemangioendothelioma, and mesenchymal hamartoma. A case report of hepatoblastoma with its attendant radiologic findings is presented. Because the prognosis is dependent on whether the tumor is surgically resectable, the accurate radiologic evaluation of patients with hepatoblastoma is critical. The roles of plain film radiography, ultrasound, computed tomography, and angiography in the evaluation of pediatric hepatic neoplasms are discussed. PMID- 2549140 TI - Occult intracellular calcium pools: relevance to neutrophil oxidant production. AB - Human neutrophils use divalent cations for the stimulus-response coupling leading to the respiratory burst. We have studied the relevance of the extracellular versus intracellular calcium stores necessary for superoxide generation in neutrophils and neutrophil cytoplasts stimulated by N-formyl-methionyl-leucyl phenylalanine (FMLP) and phorbol ester and our findings are as follows. (1) FMLP triggered superoxide anion (O2-) generation only in the presence of extracellular calcium, but phorbol myristate acetate (PMA) activation was independent of extracellular calcium. (2) inhibition of calcium transport channels with verapamil produced a dose-related inhibition of FMLP- and PMA-stimulated O2- production. Verapamil inhibition of FMLP-stimulated but not PMA-stimulated O2- was reversed simply by increasing extracellular calcium concentration. Cytosolic calcium loading with substimulatory concentrations of A23187 (0.3 to 0.6 mumol/L) (only in calcium-replete medium) could reverse the verapamil inhibition of PMA triggered O2- generation. An inhibitor of intracellular calcium availability, 3,4,5-trimethoxybenzoic acid (TMB-8), similarly inhibited FMLP-stimulated and PMA stimulated O2- production; this inhibition was reversible with A23187 calcium loading. (3) Verapamil, although it inhibited the functional oxidant response, had no inhibitory effect on fura 2-monitored cytosolic calcium rise induced by FMLP stimulation. (4) No cytosolic calcium increment was observed with PMA. O2- generation from enucleated polymorphonuclear leukocyte (PMN) cytoplasts was sensitive to inhibition by verapamil but was less so than O2- generation from intact PMNs. Cytoplasts, nearly depleted of cytoplasmic organelles and cytoskeleton, the potential sites of intracellular calcium stores, were markedly insensitive to the inhibitory effects of TMB-8.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549141 TI - In vivo threshold for cutaneous synthesis of vitamin D3. AB - Cutaneous vitamin D3 synthesis and release into the circulation is promoted by skin exposure to ultraviolet B radiation (UVB, spectrum 290 to 320 nm). To determine the relation between UVB energy level and cutaneous vitamin D synthetic response, we delivered graded increases of UVB suberythemic radiant energy (3 to 27 millijoules/cm2 [mJ/cm2]) to 32 untanned young white subjects with skin type III (Fitzpatrick-Pathak classification). Serum vitamin D3 was determined 1 hour before (basal value) and 24 hours after a single whole body exposure to UVB in a phototherapy unit. The basal vitamin D3 concentration was similar in all individuals (mean +/- SEM for whole group, 1.6 +/- 0.2 ng/ml). UVB irradiances were followed by proportional rises in serum vitamin D3 (at 27 mJ/cm2, 14.3 +/- 3.7 ng/ml), and the overall correlation between UVB radiation and consequent serum vitamin D3 response (r = 0.81; p less than 0.02) was best described by an exponential function. The minimal UVB radiation level that produced a significant increase in serum vitamin D3 was 18 mJ/cm2, a value similar to the lowest solar broadband UVB irradiance that generates previtamin D3 in vitro from the precursor 7-dehydrocholesterol (20 mJ/cm2). Because in the northern United States winter UVB irradiance does not generally reach this threshold level, we conclude that individuals living at extreme northern (or southern) latitudes may have higher dependence on body stores and dietary supply to meet their vitamin D requirements during winter. PMID- 2549142 TI - Clear cell carcinoma arising in pleomorphic adenoma of the minor salivary gland. AB - Malignant change arising in pleomorphic adenoma of the salivary glands is uncommon, with a reported incidence of 2-10 per cent. Only one case where a pleomorphic adenoma became a clear cell carcinoma appears to have been published in the English literature. An additional case of clear cell carcinoma arising in pleomorphic adenoma of a minor salivary gland is reported and the relevant literature discussed. PMID- 2549143 TI - Hepatic monoacylglycerol acyltransferase: ontogeny and characterization of an activity associated with the chick embryo. AB - Hepatic monoacylglycerol acyltransferase is expressed during the perinatal period in rats and guinea pigs and appears to be related temporally to the availability of fatty acids and to the development of hepatic steatosis. In order to determine when monoacylglycerol acyltransferase activity is expressed in an avian species, its ontogeny was investigated in chick liver total particulate preparations. In livers from 11- to 21-day-old chick embryos, monoacylglycerol acyltransferase specific activity was 34.5 +/- 8.1 nmol/min per mg of total particulate protein. The specific activity decreased 93% to 2.6 +/- 1.3 nmol/min per mg by the 6th day after hatching. The specific activities of fatty acid CoA ligase, diacylglycerol acyltransferase, and microsomal and mitochondrial glycerol-P acyltransferases changed comparatively little during this time period. In the embryos, the monoacylglycerol acyltransferase activity per liver rose 28-fold between the 11th and 21st day, corresponding exactly to the increase in liver total particulate protein during this time. Monoacylglycerol acyltransferase activity in other tissues was 25- to 115-fold lower than observed in liver. Optimal activity was measured using 25 microM palmitoyl-CoA and 50 microM sn-2-monooleoylglycerol. The activity with the 1- and 2-monooleoylglycerol ethers and 1-monooleoylglycerol was very low. In contrast to microsomes from rat liver, about 70% of the product with the 1- and 2-monooleoylglycerol ethers was triradylglycerol, suggesting that the diacylglycerol acyltransferase from chick liver can acylate acyl, alkylglycerols. The activity with sn-2-monooleoylglycerol amide was 12.5% of that observed with the corresponding 2-monooleoylglycerol suggesting that the ester bond is important; the 1-monooleoylglycerol amide was not a substrate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549144 TI - The calculation of the osmotic volumes of distribution of hypertonic sodium bicarbonate and other hypertonic solutions: a theoretical approach. AB - Correction of a base deficit with hypertonic sodium bicarbonate (NaHCO3) according to the formula 0.3 x body weight (kg) x BE may overestimate the real demands, especially during shock and cardiopulmonary resuscitation (CPR). Life threatening alkalosis and osmolal poisoning may be the consequence. As an explanation, the possibly impaired functional distribution space related to low cardiac output and volume centralization during CPR is proposed. By application of our own and outside literature, data equations are specified for an approximate precalculation of osmolality and pH changes due to infusion of hypertonic NaHCO3. Calculation of the osmotic volumes of distribution of NaHCO3 during cardiopulmonary resuscitation (CPR) from emergency literature data demonstrates a possible reduction of the functional distribution spaces of about 70 to 80% compared with normal values. This is in good agreement with the reported reduction of cardiac output to about 25% of normal during CPR. The calculated amount of sodium bicarbonate to be recommended during CPR, thus, should not exceed 50 mmol given in time intervals of at least 10 min, severe acidosis and sufficient ventilation presumed. PMID- 2549145 TI - The location of filamentous hemagglutinin and pertussis toxin antigens of Bordetella pertussis by immunoelectron microscopy. AB - Immunoelectron microscopy using colloidal gold-tagged antibodies was used to detect filamentous hemagglutinin (FHA) and pertussis toxin (PT) antigens on the surface and in the cytoplasm of Bordetella pertussis cells. Both gold-tagged antibodies to FHA and PT labeled the aggregates of filamentous material on the surface of sediment-settled phase I cells under static conditions. FHA and PT antigens were detected also on ultrathin sections made after embedding the phase I cells in Lowicryl K4M resin. On the ultrathin sections, intense label of gold tagged antibodies to FHA and PT was present on the cell surface and also in the cytoplasm, but not in the nucleoid. The aggregates of filamentous material adhering on the surface of phase I cells were most abundant on culture day 3, the end of the logarithmic growth stage, but most of the aggregates were found detached from the cell surface on culture day 5 or 7, the stationary stage. The aggregates were not found on the surface of phase III cells. The present study suggested that FHA and PT antigens were localized on the same cell structure and that both antigens were synthesized in the cytoplasm and secreted across the cell membrane mainly in the logarithmic growth stage of the phase I cells. PMID- 2549146 TI - Effect of serotonin on alpha-melanocyte-stimulating hormone secretion from perifused frog neurointermediate lobe: evidence for the presence of serotonin containing cells in the frog pars intermedia. AB - We have examined the presence of 5-hydroxytryptamine (serotonin; 5-HT) in the intermediate lobe of the frog pituitary and investigated the effect of exogenous 5-HT on alpha-melanocyte-stimulating hormone (alpha-MSH) release from the perifused neurointermediate lobe (NIL). Using a specific antiserum against 5-HT, the indirect immunofluorescence technique revealed the presence of 5-HT-like immunoreactivity (5-HT-LI) in discrete cells, generally gathered in small clusters among parenchymal cells, and in numerous neurites surrounding melanotrophic cells. At the electron microscopic level, using a silver-gold intensification procedure, 5-HT-LI was localized in dense-core secretory vesicles within specific pituitary cells which appear to be different from pituitary melanotrophs. Dense accumulation of gold particles was also observed in nerve fibres running between parenchymal cells. A combination of high-performance liquid chromatography analysis and electrochemical detection showed the presence of both 5-HT and its metabolite 5-hydroxyindol acetic acid (5-HIAA) in frog NIL extracts (534 +/- 40 and 1245 +/- 65 (S.E.M.) pg/mg wet tissue respectively). Administration of graded doses of 5-HT (from 1 to 30 mumol/l) to perifused frog NIL induced a dose-dependent inhibition of alpha-MSH release. Repeated pulses of 5-HT (10 mumol/l each) induced a reproducible inhibition of alpha-MSH without any desensitization phenomena. The inhibitory effect of 5-HT was partially blocked by the serotonergic antagonists methysergide and ICS-205-930 (10 mumol/l each). Concomitant administration of methysergide and ICS-205-930 (10 mumol/l each) totally abolished 5-HT-evoked inhibition of alpha-MSH. Fenfluramine, a releaser of 5-HT, induced a slight but significant reduction of alpha-MSH secretion. While 5-HT caused a marked inhibition of alpha-MSH release from intact NIL, 5-HT was devoid of effect on acutely dispersed pars intermedia cells suggesting that 5-HT does not exert a direct action on pituitary melanotrophs. We have examined the effect of specific dopaminergic, GABAergic and alpha-adrenergic antagonists on 5 HT-induced alpha-MSH inhibition. We observed that sulpiride and SR 95531 (10 mumol/l each) did not affect the response of NIL to 5-HT while yohimbine (10 mumol/l) suppressed the inhibitory action of 5-HT. Taken together, our results indicate that discrete cells of the frog pars intermedia contain the neurotransmitter 5-HT which may act locally to inhibit alpha-MSH release.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2549148 TI - Trypsinization of bovine parathyroid cells abolishes Ca2+-regulated parathyroid hormone secretion. AB - The secretion of parathyroid hormone (PTH) is inversely related to the extracellular Ca2+ concentration (Ca2e+). To test the hypothesis that a Ca2+ sensor on the surface of parathyroid cells is involved in Ca2+-regulated PTH secretion, limited trypsinization of bovine parathyroid cells was carried out. Treatment with trypsin (1.1-10 mg/ml) inhibited, in a dose-dependent manner, PTH secretion stimulated by lowering Ca2e+ from 2.0 to 0.5 mmol/l. In control cells, activation of protein kinase C with 12-O-tetradecanoylphorbol-13-acetate (TPA) enhanced PTH secretion at 2.0 mmol Ca2e+/l but not at 0.5 mmol Ca2e+/l. In trypsinized cells, however, TPA enhanced PTH secretion at both 0.5 and 2.0 mmol Ca2e+/l. Isoproterenol-stimulated PTH secretion was maintained in trypsinized cells, but reduced cyclic AMP production revealed that some beta-adrenergic receptors were destroyed. The cytosolic free Ca2+ concentration (Ca2i+), as measured with fura-2, was raised within seconds in response to increasing Ca2e+ from 0.5 to 2.0 mmol/l and was then lowered within 1 min to a sustained plateau; the changes were the same in trypsinized and control cells. In conclusion, trypsinization of parathyroid cells abolished Ca2+-regulated PTH secretion without affecting Ca2i+. PMID- 2549147 TI - Opioid receptors on cells of the immune system: evidence for delta- and kappa classes. AB - Opioid peptides have been shown to modulate various parameters of both the humoral and cellular arms of the immune system. The modulatory capacity of the peptides can often be substantially reduced in the presence of naloxone, an opioid receptor antagonist, indicating a classical ligand-receptor interaction. In order to characterize these interactions further, we investigated the characteristics of opioid receptors on a macrophage cell line, P388d1. A delta class opioid receptor was found with an Mr of 58,000. We also identified opioid receptors on MOLT-4 (T-cell) and IM-9 (B cell) cell lines as well as thymocytes and T cell-and B cell-enriched populations. Using the central (brain) kappa selective agonist, U-69,593, it was also determined that P388d1 cells possess kappa-like opioid receptors. Scatchard analysis of the binding of [3H]U-69,593 revealed a single population of sites with a dissociation constant of 17 +/- 3 (S.E.M.) nmol/l and a total number of binding sites of 53.8 +/- 1.0 (S.E.M.) fmol/10(6) cells. Moreover, the racemic kappa-selective agonist U-50,488H was able to displace 50% of [3H]U-69,593 binding at 8.0 nmol/l, whereas other opioid ligands such as [Met]-enkephalinamide (delta-selective) and [D-Ala2,N-Me Phe4,Gly5-ol]-enkephalin (mu-selective) were ineffective displacers of [3H]U 69,593 except at high concentrations. PMID- 2549150 TI - Isolation of the human plasma corticotrophin-releasing factor-binding protein. AB - We report the purification of human corticotrophin-releasing factor-binding protein (hCRF-BP) using repeated affinity chromatography (with the aid of synthetic CRF immobilized on Sepharose 4B solid phase) followed by gel filtration. Presence of the binding protein was tracked throughout the procedure by its ability to inhibit binding of 125I-labelled hCRF to an hCRF antiserum; normal human plasma exhibits 80% inhibition in this system whereas sheep plasma, which does not contain an hCRF-BP, has no effect. Affinity cross-linking of 125I labelled hCRF to the purified hCRF-BP was performed using disuccinimidyl suberate (1 mmol/l). SDS electrophoresis of the purified CRF cross-linked binding protein followed by radioautography resulted in one major band of Mr 37,000 which corresponded to our original molecular weight estimate based on the elution position of the binding protein on Sephacryl S-200. A 10(7)-fold purification of the hCRF-BP resulted in a preparation estimated to be 95% pure, with an overall yield of 5%. PMID- 2549149 TI - Actions of parathyroid hormone-related protein on the rat kidney in vivo. AB - Peptides containing residues 1-34 of parathyroid hormone-related protein (PTHrP) and of bovine parathyroid hormone (bPTH), and recombinant full-length PTHrP(1 141) were infused i.v. into anaesthetized thyroparathyroidectomized rats to compare their action and potency on the renal handling of calcium, phosphate and cyclic AMP (cAMP) in vivo. All three peptides decreased the excretion of calcium and increased the excretion of phosphate and cAMP in the urine, with PTHrP(1-34) and PTHrP(1-141) having virtually equipotent effects. Thus the essential requirements for the major physiological activity of PTHrP on the kidney are contained within the 34 amino-terminal amino acids. For all three peptides, the lowest infusion rate that increased phosphate and cAMP excretion was 0.01 nmol/kg per h, whereas the lowest infusion rate that decreased calcium excretion was 0.025 nmol/kg per h for the PTHrP peptides and 0.1 nmol/kg per h for bPTH(1-34). The response to the PTHrP peptides was maximal at an infusion rate of 0.1 nmol/kg per h for both calcium and phosphate. Since the kidney is either equally sensitive to PTHrP and bPTH(1-34), or more sensitive to PTHrP than to bPTH(1-34), the hypercalcaemia of humoral hypercalcaemia of malignancy may develop because uncontrolled secretion of PTHrP increases the renal reabsorption of calcium to such an extent that even a modest increase in the inflow of calcium into the blood raises plasma calcium concentration. PMID- 2549151 TI - Effects of recombinant human interleukin-1 alpha, -1 beta, 2 and 6 on ACTH synthesis and release in the mouse pituitary tumour cell line AtT-20. AB - The effects of recombinant human interleukin (rhIL)-1 alpha, -1 beta, 2 and 6 on the release of ACTH from the ACTH-producing tumour cell line AtT-20 of the mouse were studied during relatively long periods of incubation. Levels of ACTH in the media, measured by radioimmunoassay, were increased by the addition of rhIL-1 alpha or -1 beta after latent periods of more than 4 h. RhIL-1 alpha and -1 beta were almost equally potent in this experiment and the minimum, half-maximum and maximum effective concentrations of both rhIL-1 alpha and -1 beta were about 0.1 pmol/l, 1-3 pmol/l and 10-100 pmol/l respectively. During incubation with rhIL-1 beta, immunoreactive ACTH levels and mRNA levels of the ACTH precursor pro opiomelanocortin in cells also increased without apparent changes in the growth rate of the cells. Although the AtT-20 cells used in this study were quite insensitive to human/rat corticotrophin-releasing hormone (CRH), the cells showed a significant response to CRH after incubation with rhIL-1 beta. RhIL-6 showed similar effects to those of rhIL-1 beta on ACTH synthesis and release; increasing ACTH in cells and media after a certain latent period. On the other hand, rhIL-2 did not change ACTH levels in the AtT-20 cells in this study. These observations indicate that rhIL-1 alpha, -1 beta and rhIL-6 have direct effects on ACTH producing cells to stimulate the release and synthesis of ACTH after a latent period. PMID- 2549153 TI - Effect of sodium intake on phosphoinositides and inositol trisphosphate response to angiotensin II, K+ and ACTH in rat glomerulosa cells. AB - To assess the impact of sodium intake on the adrenal phosphoinositide system, rats were maintained on a low or normal salt diet for 5 days, and glomerulosa cell preparations (2 x 10(5) cells) were stimulated by angiotensin II (AII; 10 nmol/l), potassium (K+; 8.7 mmol/l) or ACTH (0.1 nmol/l) for 0, 2, 4, 6, 12, 15 and 60 s. Levels of phosphatidylinositol (PtdIns), phosphatidylinositol 4 phosphate (PtdIns 4-P), phosphatidylinositol 4,5-bisphosphate (PtdIns 4,5-P2) and inositol 1,4,5-trisphosphate (Ins 1,4,5-P3) + inositol 1,3,4-trisphosphate (Ins 1,3,4-P3) were assayed by a microspectrophotometric procedure. Non-stimulated levels of PtdIns, PtdIns 4-P, PtdIns 4,5-P2 and Ins 1,4,5-P3 (+ Ins 1,3,4-P3) (means +/- S.E.M.; n = 36) in cells from rats on the low Na+ intake were 580 +/- 6.5, 187 +/- 2.6, 82 +/- 3 and 95 +/- 1.2 pmol per incubate respectively, indistinguishable from those observed in rats on a normal Na+ intake, except for the significantly (P less than 0.025) greater PtdIns 4,5-P2 level. In response to AII stimulation, all four compounds showed an earlier and greater peak response when cells were obtained from animals on a low rather than a high sodium intake. All values has returned to control levels by 12-15 s, regardless of the level of sodium intake. In contrast, with K+ stimulation there were no differences in the peak response of cells from rats on the two dietary intakes, but there was a shift of the peak to a longer time-interval (6 versus 8 s) in animals maintained on a low sodium intake.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549152 TI - Angiotensin II and potassium activate different calcium entry mechanisms in rat adrenal glomerulosa cells. AB - Initial 45Ca uptake was measured in isolated rat glomerulosa cells. A small reduction in membrane potential produced by increasing the K+ concentration from 2 to 3.6 mmol/l stimulated 45Ca uptake by about 35%, while a bigger depolarization induced by 18.5 mmol K+/l increased the uptake by about 100%. Since Ca2+ influx was already activated at a calculated membrane potential below 70 mV, and was found to be sensitive to the dihydropyridine antagonist nifedipine (1 mumol/l), but insensitive to nickel ions (100 mumol/l), it does not meet the criteria established for T- or L-type voltage-dependent Ca2+ channels. Exposure of glomerulosa cells to angiotensin II (AII) for 10 min also enhanced the rate of 45Ca influx. The effect of AII was not sensitive to 1 mumol nifedipine/l, but was strongly inhibited by 5-(N-4-chlorobenzyl)-N-(2',4'-dimethyl)benzamil (CBDMB, 30 mumol/l), an inhibitor of the Na+/Ca2+ antiporter. These observations suggest that during the sustained phase of stimulation with AII, a CBDMB-sensitive mechanism, rather than dihydropyridine-sensitive calcium channels, is involved in Ca2+ uptake in rat glomerulosa cells. The bulk Ca2+ influx did not correlate with aldosterone production; however, the maintained activity of different Ca2+ entry mechanisms seems to be essential for AII-induced aldosterone production. PMID- 2549154 TI - Increased levels of a humoral digitalis-like factor in deoxycorticosterone acetate-induced hypertension in the pig. AB - Plasma levels of an endogenous digitalis-like factor (EDLF) and atrial peptides were followed in pigs confined to metabolic cages during the development of deoxycorticosterone acetate (DOCA)-induced hypertension. During the first 2 days of DOCA treatment, urinary sodium excretion decreased and the plasma levels of renin and atrial peptide fell significantly. During this period, plasma levels of EDLF increased greater than 30-fold from a baseline of less than 0.25 to 9.72 nmol ouabain equivalents/l. Between days 2 and 5 of DOCA treatment, urinary sodium returned to pre-DOCA levels ('mineralocorticoid escape') and during this period significant increases of atrial peptide and mean arterial pressure (MAP) and a decrease in EDLF were found. Following mineralocorticoid escape there was a secondary rise in levels of EDLF and atrial peptide and both phenomena correlated with MAP (EDLF, r = 0.87, P less than 0.05; atrial peptide, r = 0.9, P less than 0.05) and with each other (r = 0.96, P less than 0.05) over a 20-day period. Acute expansion of extracellular fluid volume before DOCA elicited significant increments in plasma EDLF and atrial peptide. Volume loading during chronic DOCA treatment increased plasma EDLF significantly whereas no response of atrial peptide was detected. These results suggest that DOCA affects the reactivity of mechanisms involved in the perception of and/or response to acute changes in volume status. However, neither EDLF nor atrial peptide appear to be viable candidates as direct mediators of mineralocorticoid escape. Finally, the nature of the changes found in EDLF and atrial peptide levels during DOCA treatment suggest that these factors are involved in the long-term control of blood pressure in this model of low renin hypertension. PMID- 2549155 TI - Feedback control of vasopressin and corticotrophin secretion in conscious dogs: effect of hypertonic saline. AB - Glucocorticoids are known to inhibit the ACTH response to a variety of stimuli. It has been suggested that vasopressin secretion is also inhibited by glucocorticoid negative feedback. The purpose of this study was to (1) determine the ACTH response to hypertonic saline and its sensitivity to glucocorticoid negative feedback and (2) to determine whether physiological elevations of plasma cortisol inhibit subsequent vasopressin responses to hypertonic saline. Five mongrel dogs (15-18 kg) were prepared with chronic arterial and venous catheters and studied while conscious. Ten experiments were performed on each dog in a randomized design separated by at least 5 days. Each experiment consisted of a pretreatment period (from -60 to -30 min except for dexamethasone administration) during which a glucocorticoid feedback signal was applied and a stimulus period (from 0 to 30 min) during which hypertonic saline was infused. The pretreatment and stimulus periods were separated by 30 min. Pretreatments were as follows: isotonic saline (control), half-maximal and maximal cortisol infusion (5.5 or 11 nmol/kg per min), ACTH(1-24) infusion (6.8 pmol/kg per min) which produces increases in endogenous cortisol, and dexamethasone (1.5 mg i.m.) given at 17.00 h the day before experimentation. Stimuli were as follows: hypertonic saline was infused at 0.2 or 0.4 mmol/kg per min which increased plasma sodium by about 6 or 12 mmol/l respectively. NaCl infusion at 0.2 mmol/kg per min had no effect on plasma ACTH or cortisol except when subsequent to ACTH(1-24) pretreatment when plasma ACTH actually increased to 41.4 +/- 2.9 pmol/l in response to hypertonic saline.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549156 TI - Growth hormone (GH) release after administration of GH-releasing hormone in relation to endogenous 24-h GH secretion in short children. AB - Endogenous GH secretion was measured every 20 min for 24 h in 36 short children. This was immediately followed by an i.v. injection of GH-releasing hormone (GHRH)(1-29)-NH2 (1 microgram/kg), and GH was estimated every 15 min for the following 2 h. The aim was to determine whether endogenous pulsatile GH secretion had any relation to, or influence on, the GH release induced by GHRH. A high variability was found both in the 24-h GH secretion expressed as area under the curve above the baseline (0-1588 mU/l x 24 h) and the maximal GH response to GHRH (5-296 mU/l), as well as after an arginine-insulin tolerance test (4-59 mU/l). We found a positive correlation (correlation coefficient of Spearman (rs) = 0.49; P less than 0.01) between the GH response to GHRH and the spontaneous GH secretion over a 24-h period, in spite of a negative correlation (rs = -0.80; P less than 0.01) with the GH secretion during the preceding 3 h. We conclude that the GH response to a GHRH test correlates with endogenous GH secretion in short children, and may be helpful in estimating the ability to release GH. It is important, however, to be aware of the influence of the spontaneous GH secretion during the 3 h immediately preceding administration of GHRH. PMID- 2549157 TI - The presence of beta-adrenoceptors in rat endometrium is dependent on circulating oestrogen. AB - It has been proposed that the change in sensitivity of the uterus to catecholamines during the oestrous cycle may result from changes in beta adrenoceptor density in the myometrium. In the present work we applied the technique of light-microscopic autoradiography using the radioligand [125I]iodocyanopindolol to study the density and distribution of beta adrenoceptors in the rat uterus during the normal oestrous cycle, following ovariectomy, and after ovariectomy with oestradiol replacement. We found that beta-adrenoceptor density in the endometrium, but not the myometrium, varied during the oestrous cycle, being highest during pro-oestrous. The endometrium of the ovariectomized rats was devoid of any beta-adrenoceptors, whilst that of the oestradiol-treated rats was very intensely labelled. Following ovariectomy there was an increase in beta-adrenoceptor density in the myometrium, but this was not observed in the animals given oestradiol replacement. It appears that both the presence and density of beta-adrenoceptors in rat endometrium is dependent on circulating oestrogens. PMID- 2549158 TI - Stress-related peptide hormones in the placenta: their possible physiological significance. PMID- 2549159 TI - Characterization of oxytocin receptors in rat adenohypophysis using a radioiodinated receptor antagonist peptide. AB - Oxytocin may function as a hypothalamic releasing hormone for prolactin and ACTH secretion in the rat. In the present study we have investigated the properties of putative oxytocin receptors in the rat adenohypophysis by radioligand-binding assay. A novel oxytocin receptor antagonist [1-(beta-mercapto-beta,beta cyclopentamethylene propionic acid),2-(ortho-methyl)-Tyr2-Thr4-Orn8-Tyr9-NH2] vasotocin (OTA) was radioiodinated by the iodogen method to a specific activity of 0.6 nCi/fmol. The radioiodinated derivative 125I-labelled OTA (125I-OTA) was reacted with membrane suspensions prepared from the uterus or adenohypophysis of female rats which were (a) ovariectomized for 7 days, (b) ovariectomized and treated with 5 micrograms oestradiol-17 beta 48 h before death or (c) implanted with a piece of silicone elastomer tubing containing 50 mg diethylstilboestrol (DES) 5 days before death. In uterine as well as the pituitary membrane suspensions, the radioligand was bound reversibly and with high affinity (dissociation constants 0.2 +/- 0.1 and 0.1 +/- 0.01 nmol/l respectively; mean +/ S.E.M., n = 3) to a single class of sites with limited binding capacity, which varied with the type of pretreatment. Oestradiol-17 beta increased the binding capacity fivefold in the uterus in ovariectomized rats, but only very low specific radioligand binding was found in pituitary preparations from the same animals. Treatment with DES markedly increased the number of receptors in both the uterus and the adenohypophysis. Studies with several agonist and antagonist analogues revealed no difference in the ligand specificity of the uterine and adenohypophysial sites binding 125I-OTA, indicating that they are the same species of receptor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549160 TI - Burmese Russell's viper venom causes hormone release from rat pituitary cells in vitro. AB - Acute and chronic hypopituitarism is associated with severe envenoming by the Burmese Russell's viper. We have demonstrated that in vitro, Burmese Russell's viper venom (0.1-10 micrograms/ml) causes a dose-dependent release of GH, TSH and ACTH from dispersed rat anterior pituitary cells in culture. At 10 micrograms/ml, venom causes a significant increase in the release of GH (344%, P less than 0.001), TSH (168%, P less than 0.005) and ACTH (greater than 700%, P less than 0.001). We have also shown that the component (or components) responsible for this stimulatory effect is stable to heat (60 degrees C, 1 h) and mild trypsinization. Repeated addition of venom (1 microgram/ml) to pituitary cells in a perifusion column system demonstrated attenuation of GH release. This reduced response was not due to depletion of the GH pool since the pituitary cells were subsequently able to respond to both GH-releasing factor (GRF) stimulation and KCl depolarization. Somatostatin in a dose which abolished GRF-stimulated GH release failed to affect venom-stimulated GH release, implying that venom acts in a cyclic AMP-independent manner. We conclude that Burmese Russell's viper venom has direct effects on pituitary hormone release in vitro. Whether these effects contribute to its known actions in vivo on the function of the pituitary remains to be established. PMID- 2549161 TI - [3H]angiotensin II binding to basolateral membranes from rat proximal renal tubule: effect of sodium intake and captopril. AB - Basolateral membranes were prepared from rat renal cortex by density gradient centrifugation. Their purity was confirmed by electron microscopy and by marker enzyme enrichment. The basolateral membrane preparation was shown to be derived predominantly from the proximal renal tubule by measurement of hormone-stimulated adenylate cyclase; marked stimulation of adenylate cyclase was found with parathyroid hormone, but not with isoprenaline, antidiuretic hormone or calcitonin. A single class of specific high-affinity [3H]angiotensin II-binding site was identified in the basolateral membrane preparation which, after correction of results for tracer degradation, showed equilibrium dissociation constant of 0.23 nmol/l and binding site concentration of 485.8 fmol/mg protein. Binding sites for [3H]angiotensin II were measured in basolateral membranes prepared from rats fed diets with a low, normal or high sodium content. A trend of increased binding site density with reduced sodium intake was found which did not reach statistical significance. No effect on affinity was demonstrated. Treatment of rats on a low-sodium diet with captopril (500 mg/l drinking water) caused a significant reduction in binding site density; no effect on affinity was demonstrated. These findings suggest that the density of angiotensin II receptors at this site is altered by the activity of the renin-angiotensin system. PMID- 2549162 TI - Stimulatory effect of caffeine on the hypothalamo-pituitary-adrenocortical axis in the rat. AB - Intraperitoneal injection of caffeine (12.5-100 mg/kg) into rats caused a significant, dose-related increase in plasma corticosterone 2 h later, when the greatest response was measured. The corticosterone response to laparotomy stress or i.v. injection of ACTH(1-24) was unaffected by prior injection of caffeine. The response to stress or caffeine was unaffected by adrenal enucleation 28 days previously. In vitro, 10 mmol caffeine/l stimulated basal release of corticosterone from adrenal quarters and potentiated the response to a sub maximal stimulatory concentration of cyclic AMP (cAMP). The drug had no effect on release stimulated by a sub-maximal concentration of ACTH(1-24). Release of ACTH from pituitary fragments incubated in vitro was stimulated in a dose-related manner by caffeine (0.01-10 mmol/l), and the responses to hypothalamic extract and sub-maximal concentrations of corticotrophin-releasing factor (CRF-41) or arginine vasopressin (AVP), but not cAMP, were significantly enhanced by 10 mmol caffeine/l. Release of immunoreactive CRF-41 (but not AVP) was significantly increased by caffeine (0.01-10 mmol/l) added to hypothalami incubated in vitro. The response to injection of caffeine in vivo was completely prevented by pharmacological blockade of endogenous CRF release. Taken together, these results show that caffeine at high concentrations can stimulate directly the release of the hormones of the hypothalamo-pituitary-adrenocortical axis in vitro, but the fact that these concentrations are unlikely to be reached after administration in vivo suggests that the effect of caffeine may be mediated centrally. PMID- 2549163 TI - Glucocorticoids act rapidly in vitro to attenuate second messenger responses to ACTH secretagogues in rats. AB - Fragments of rat anterior pituitary glands incubated in vitro and challenged with either of two ACTH secretagogues were used to investigate the extent to which the acute, biphasic, feedback-like inhibitory effects on hormone secretion exerted by the synthetic glucocorticoid dexamethasone were related to alterations in second messenger responses. Addition of dexamethasone was shown to cause both an immediate inhibition (fast inhibition) of the release of ACTH-like immunoreactivity induced by arginine vasopressin (AVP) or corticotrophin releasing factor (CRF-41), and also an inhibition that occurred after removal of the steroid and was maximal 90 min after its introduction (early delayed inhibition). The accumulation of adenosine 3',5'-monophosphate (cAMP) in the tissue was enhanced in a dose-related manner by CRF-41, as was that of phosphate esters of inositol (inositol phosphates) by AVP. The dose-response curve for the effect of CRF-41 on cAMP production was markedly shifted to the right by dexamethasone acting in the time-domain of fast inhibition (i.e. the response was attenuated, but not abolished). Application of the steroid during the same time period reduced significantly the inositol phosphate response induced by the higher concentration of AVP tested (3000 nmol/l), but had no effect on the action of a lower concentration (30 nmol/l). In contrast, the cAMP and inositol phosphate dose-response curves to CRF-41 and AVP respectively were unaffected by the glucocorticoid when it was applied at the time which generated early delayed inhibition of ACTH release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549164 TI - Presynaptic calcium currents and facilitation of serotonin release at synapses between cultured leech neurones. AB - 1. The role of presynaptic Ca2+ entry in facilitation of transmitter release has been analysed by voltage-clamp measurements at synapses formed in culture by Retzius and P neurones isolated from the central nervous system (CNS) of the leech. The transmitter released by Retzius cells is serotonin. 2. Synaptic transmission persisted in solutions containing raised concentrations of divalent cations, reduced concentrations of Na+, and tetraethylammonium (TEA+) and 4-AP (to block K+ currents). Ca2+ and Sr2+ were more effective in promoting transmitter release than Ba2+, as assessed by the postsynaptic potentials in P cells. The degree and time course of facilitation in Ca2+- and Sr2+-containing solutions were similar to those observed for synapses bathed in normal L-15 medium. 3. Transmitter release depended upon the amplitude and the duration of presynaptic depolarization and inward Ca2+ current. Peak Ca2+ currents and postsynaptic potentials occurred with depolarizing steps to +15 mV. Frequent or prolonged pulses depressed the postsynaptic potentials. 4. Pairs of depolarizing pulses that caused facilitation were accompanied by identical inward Ca2+ currents. These results indicate that the mechanism responsible for facilitated serotonin release must occur following Ca2+ entry and that residual Ca2+ plays a role. PMID- 2549165 TI - Metabolism of isolated fish gill cells: contribution of epithelial chloride cells. AB - Gill cell suspensions from freshwater (FW)- and seawater (SW)-adapted teleosts were obtained by density gradient centrifugation. The proportion of chloride cells (CCs) in the mixed cell suspensions was estimated using the fluorescent mitochondrial stain, DASPMEI, and ranged from less than 1% (FW-adapted tilapia) to approximately 13% (SW-adapted toadfish). The gill cells displayed relatively high viability based on Trypan Blue exclusion (greater than 75%), lactate dehydrogenase leakage (less than 6.5% h-1), oxygen consumption rates (5-15 mumol g-1 cell wet mass h-1) and ATP levels (1-3 mumol g-1 cell wet mass). There were no obvious differences between the viability of CCs and the other cell types present. An initial comparison of gill oxidative metabolism in SW-adapted tilapia (Oreochromis mossambicus) and toadfish (Opsanus beta) demonstrated that both species oxidized glucose and lactate at substantially greater rates than alanine or oleate. Metabolic rates were significantly higher in toadfish cell suspensions. Kinetic experiments revealed that toadfish gill cells displayed lower values of Km and higher values of Vm for both lactate and glucose, in comparison to tilapia. The elevated metabolism in toadfish gill cells was correlated with increased activities of the oxidative enzyme citrate synthase and Na+/K+-ATPase. The toadfish cell suspensions had a greater proportion of CCs and it is likely that the difference in CC numbers between the two species is the basis for the observed differences in enzyme activities and rates of oxidative metabolism. This idea is supported by the highly significant correlation between Na+/K+-ATPase activity (or CC numbers) and rates of lactate oxidation in gill cell suspensions from FW- and SW-adapted tilapia and toadfish, as well as SW adapted tilapia chronically treated with cortisol to elevate CC numbers. Although it has been assumed widely that the high metabolic rate of gill tissue reflects, in part, the oxidative demands of the chloride cell, the results of this study provide the first experimental, albeit indirect, evidence for differential rates of metabolism in the various cell types that comprise the gill. PMID- 2549166 TI - The monocyte-derived neutrophil activating peptide (NAP/interleukin 8) stimulates human neutrophil arachidonate-5-lipoxygenase, but not the release of cellular arachidonate. AB - LPS and mitogen-stimulated mononuclear cells secrete a cytokine, which is able to activate the PMNL-arachidonate-5-lipoxygenase. This cytokine has been proven to be identical with the recently characterized novel neutrophil-activating peptide NAP/IL-8. NAP/IL-8 is able to activate human PMNL for release of LTB4, omega oxidized LTB4, and 5-HETE in the presence of exogenous AA. Half-maximal concentration of NAP/IL-8 for release of LTB4 has been found to be near 4 x 10( 8) mol/liter. Time course studies revealed rapid activation of PMNL, with maximal release of LTB4 within the first 10 min with a decline up to 40 min. High amounts of omega-oxidized LTB4 were detected up to that time. Significant amounts of AA-5 LO-products can be detected only when PMNL were stimulated with NAP/IL-8 in the presence of exogenous AA. The concentration of AA necessary for half-maximal LTB4 release has been found to be 3 x 10(-6) mol/liter. In the presence of 8 x 10(-9) mol/liter [3H]AA, NAP/IL-8 (10(-9) to 10(-7) mol/liter) did not induce the production of LTB4, omega-oxidized LTB4, or 5-HETE. In addition, PMNL prelabeled with [3H]AA did not release either [3H]AA or 5-lipoxygenase metabolites when stimulated with NAP/IL-8 (10(-9) to 10(-7) mol/liter), indicating that NAP/IL-8 apparently does not activate cellular phospholipases/diacylglycerol-lipases. Apart from FMLP, C5a, and PAF NAP/IL-8 is the fourth clearly characterized neutrophil chemotaxin able to activate the PMNL-5-lipoxygenase. The detection of large amounts of NAP/IL-8, arachidonic acid, as well as LTB4-like material, in lesional material of patients with psoriasis points towards a possibly important role of NAP/IL-8 in amplifying inflammatory processes by induction of LTB4 production. PMID- 2549167 TI - Antibody inhibits defined stages in the pathogenesis of reovirus serotype 3 infection of the central nervous system. AB - The mammalian reoviruses provide a model for studying specific aspects of the immunopathogenesis of viral infection. We have used two serotype 3 reoviruses to define stages in the pathogenesis of central nervous system (CNS) infection at which a mAb specific for the reoviral cell attachment protein sigma 1 (sigma 1mAbG5) acts to protect mice against lethal disease. sigma 1mAbG5 administered either before or at the time of footpad inoculation with reovirus T3D prevented entry of T3D into the CNS. sigma 1mAbG5 also inhibited the spread of reovirus T3C9 from the gastrointestinal tract to the CNS after peroral inoculation with T3C9. These effects occurred in the absence of a significant effect of sigma 1mAbG5 on primary replication in skeletal muscle (T3D) or the gastrointestinal tract (T3C9). sigma 1mAbG5 administered after T3D had reached the spinal cord inhibited subsequent spread of infectious virus from spinal cord to brain. Even after direct intracerebral inoculation of T3D, sigma 1mAbG5 prevented both growth in the brain and spread of infectious virus from brain to eye, spinal cord, and muscle. Treatment with sigma 1mAbG5 after intracerebral inoculation with T3D prevented neuronal necrosis and resulted in a delayed and topographically restricted inflammatory response. We detected no antibody-resistant T3D variants in vivo after treatment with sigma 1mAbG5. We conclude that systemic IgG does not play a significant role at the primary site of infection with reoviruses, while it clearly acts to prevent infection of the CNS and extension of infection with the CNS. Further study will be directed to defining what components of the immune system do act at primary sites of infection, and to defining the mechanisms by which antibody acts at defined stages in pathogenesis. PMID- 2549168 TI - Antagonistic control of tumor necrosis factor receptors by protein kinases A and C. Enhancement of TNF receptor synthesis by protein kinase A and transmodulation of receptors by protein kinase C. AB - We have investigated control mechanisms of TNF receptor expression (TNF-R) in various human tumor cells and normal peripheral blood monocytes. Activators of protein kinase A (PKA) signal transduction pathways were found to enhance TNF-R expression up to sevenfold, whereas in the same cells, IFN-alpha and -gamma receptors remained unaffected. Inhibitors of protein kinases downregulate both constitutive and cAMP-enhanced TNF-R expression. Binding studies revealed an increase in TNF-R numbers without a change in receptor affinity. Both, direct activators of PKA and inhibitors of phosphodiesterase, raising intracellular levels of cAMP, were found to be effective. As activation of PKA does not slow down the degradation rate of TNF-Rs, but rather enhances protein synthesis dependent reexpression of TNF-Rs after transient PKC-mediated transmodulation and after tryptic digestion of TNF-Rs, it is concluded that PKA stimulates TNF-R synthesis. Maximum TNF-Rs enhancement is reached after 24 h of stimulation and is reversible, suggesting that receptor upregulation is not linked to irreversible steps of cellular differentiation. PKA-mediated enhancement of TNF-R expression was predominantly observed in normal peripheral blood monocytes and tumor cell lines of myeloid origin. As in these typical TNF producer cells, the production of TNF is also controlled by PKA and PKC, a regulatory circuit is proposed, by which these two independent signal pathways antagonistically regulate TNF production and, at the receptor level, TNF sensitivity. PMID- 2549169 TI - Energy transport and cell polarity: relationship of phosphagen kinase activity to sperm function. AB - The energy required for motility of sea urchin sperm is transported from the mitochondrion to the flagellum by a phosphocreatine shuttle involving diffusion of phosphocreatine (PCr) between isozymes of creatine kinase (CrK) localized at the two sites (Tombes and Shapiro, Cell, 41:325, '85; Tombes et al., Biophys. J., 52:75, '87). The present studies demonstrate that high sperm CrK (various echinoderms; sea squirt, bristle worm, salmon) or arginine kinase (molusc, barnacle, moth) activity is seen in several species with sperm of a primitive morphology (mitochondrion at the base of the head, relatively long flagellum). In contrast, CrK activity is 10-100-fold less abundant in sperm of other species (frog, mouse, rooster, rabbit, bull, and human) that either possess a modified morphology (mitochondria that extend along the flagellum) and/or utilize glycolytic metabolism. We interpret these findings as support for the use of phosphagen kinase-dependent energy transport in cells in which the production of adenosine triphosphate (ATP) by the mitochondrion is distant from its utilization, leading to a form of metabolic polarization. Two other cell types, frog photoreceptors and rabbit oviduct cells, whose morphology and function also suggest that they exhibit metabolic polarization, contain relatively high CrK activity. The presence of high phosphagen kinase activity in metabolically polarized gametes and somatic cells further substantiates the role of such enzymes in facilitating energy transport. PMID- 2549170 TI - Plasminogen activator activity is associated with neural crest cell motility in tissue culture. AB - We have examined the possibility that proteases such as plasminogen activator (PA) contribute to the extraordinary motile capability of neural crest cells. We show that trunk neural crest cells that migrate from isolated neural tubes in vitro produce PA and that the level of cell-associated PA increases dramatically after 8 days in culture. This increase is not the result of differentiation or time in culture, because neural crest cell clusters that form on top of the neural tube and differentiate into pigment cells but are immotile produce very low levels of PA. If these clusters are removed from the neural tube and replated on a plastic substratum where they migrate, the level of PA associated with the cells increases dramatically, suggesting that PA production is associated with motility. Inhibitors of PA/plasmin activity significantly reduce neural crest cell motility in vitro, further supporting the idea that proteases are important in neural crest cell migration. PMID- 2549172 TI - Comparison of histological effects of different bioceramic implants on surgically created periodontal defects in monkeys. AB - The biological response to the resorbable and nonresorbable bone grafting materials implanted in the surgically created defects was compared in 4 Maccaca cyclopsis. Intraosseous angular defects were created on the mesial side of both upper canines and premolars by an air driven fissure bur. The root surface was planed, then hydroxylapatite (HA) and tricalcium phosphate (TCP) were inserted into the right side defects respectively. The left side canine received no implants and served as the control site. The animals were sacrificed 6 and 16 weeks after the experiment. Histologically, the two implant materials were well tolerated by the surrounding tissues and did not elicit any adverse inflammatory reaction. During the 16 weeks periods, TCP was biodegraded through phagocytosis by histiocytes and multinucleated giant cells. The residual particles were incorporated into the new bone matrix and displayed the bone growth guiding property. HA particles were dense and larger in size with low resorption rate, therefore, encapsulation by fibrous tissue was more commonly seen. Although direct bone deposition adjacent to the HA particle surface was apparently found, new bone formation was observed only at the apical portion adjacent to the host alveolar bone. These findings suggested that HA might rather play the role as a biocompatible filler. However, HA was more effective in blocking the downgrowth of the junctional epithelium. Nevertheless, these two implant materials could not hasten the healing process, because new bone formation was observed to be more distinct in the control site. PMID- 2549171 TI - Proteolytic activity of specialized surface protrusions formed at rosette contact sites of transformed cells. AB - Surface protrusions at the leading edge of a moving cell that make contact with the surrounding extracellular matrix (ECM) are its main motor for locomotion and invasion. Chicken embryonic fibroblasts transformed by Rous sarcoma virus (RSV CEF) form specialized membrane rosette-shaped contact sites on planar substrata as shown by interference reflection microscopy (IRM). Such activity is lacking in normal cells. These rosette contacts are more labile than other adhesion sites, such as focal and close contacts. Ultrastructural studies demonstrate that rosettes are sites at which membrane protrusions from the ventral cell surface contact the substratum. These protrusions are filled with meshworks of microfilaments and contain the pp60src oncogene product, actin, vinculin, and alpha-actinin. However, unlike focal contacts, at the rosettes these proteins interact to extend a highly motile membrane. Rosettes have the biological activity of degrading ECM components, as demonstrated by (1) local degradation of fibronectin substrata at sites of rosette contacts, but not focal and close contacts; (2) localization of putative antiprotease antibody at sites of rosette contacts, but not at focal an close contacts; and (3) local disruption of fibronectin matrix at sites of protrusive activity seen by transmission electron microscopy (TEM). In addition, formation of the rosette contact is insensitive to the ionophore monensin, and to inhibitors of proteolytic enzymes, while local fibronectin degradation at rosette contacts is inhibited by inhibitors of metalloproteases, 1,10-phenanthroline and NP-20. I consider these membrane protrusions of the rosette contacts in RSV-transformed cells specialized structural entities--invadopodia--that are involved in the local degradation of the ECM. PMID- 2549173 TI - [Comparative study of antihypertensive effects of enalapril and atenolol in patients with mild to moderate hypertension]. PMID- 2549174 TI - Tonic and phasic block of neuronal sodium currents by 5-hydroxyhexano-2',6' xylide, a neutral lidocaine homologue. AB - The effects of a neutral lidocaine homologue, 5-hydroxyhexano-2',6'-xylidide (5 HHX), on the kinetics and amplitude of sodium currents in voltage-clamped amphibian nerve fibers are described. 5-HHX produced two types of sodium current inhibition: (a) tonic block, in resting fibers (IC50 approximately 2 mM), and (b) phasic block, an additional, incremental inhibition, in repetitively depolarized fibers (frequency greater than 1 Hz). The kinetics of phasic block were characterized by a single-receptor, switched-affinity model, in which binding increases during a depolarizing pulse and decreases between pulses. In the presence of 4 mM 5-HHX, binding increased during pulses from -80 to 0 mV, with an apparent rate constant of 6.4 +/- 1.4 s-1. Binding decreased between pulses with an apparent rate constant of 1.1 +/- 0.3 s-1. There was little effect of extracellular pH on the kinetics of phasic block. These findings demonstrate that neither the presence of a terminal amine nor a net charge on a local anesthetic is required for phasic block of sodium channels. PMID- 2549175 TI - Calcium dependence of the activation and inactivation kinetics of the light activated phosphodiesterase of retinal rods. AB - The Ca2+ dependence of the kinetics and light sensitivity of light-activated phosphodiesterase was studied with a pH assay in toad and bovine rod disk membranes (RDM), and in a reconstituted system containing GTP-binding protein, phosphodiesterase and rhodopsin kinase. Three statistics, peak hydrolytic velocity, turnoff time, and time to peak velocity, were measured. ATP decreased phosphodiesterase light sensitivity nearly 10-fold and accelerated the dim-flash kinetics of cGMP hydrolysis when compared to those with GTP alone. CA2+ reversed all of the effects of ATP, Ca2+ increased peak velocity, turnoff time, and time to peak velocity, to the values obtained with GTP alone. The Ca2+ dependence of peak velocity and turnoff time can be characterized as hyperbolic saturation functions with a K0.5 for Ca2+ of 1.0-1.5 mM in toad RDM. In bovine RDM the Ca2+ dependence of peak velocity and turnoff time has a K0.5 of 0.1 mM Ca2+. The Ca2+ dependence in the reconstituted system is similar to that in bovine RDM for peak velocity (K0.5 = 0.1 mM Ca2+) but differs for turnoff time (K0.5 = 2.5 mM Ca2+). We tested the hypothesis that a soluble modulator, normally required to confer submicromolar Ca2+ sensitivity, was too dilute in our assay by comparing data obtained at one RDM concentration with those obtained at 10-fold higher RDM, and therefore a constituent protein, concentration. We observe no difference and present a formal analysis of these data that excludes the hypothesis that the soluble modulator binds its target protein with Kd less than 5 microM. The lack of submicromolar Ca2+ dependence of any of the steps in the cGMP cascade that underlie cGMP phosphodiesterase activation and inactivation in vitro argues against Ca2+ regulation of these steps having a significant role in the light adaptation of the intact rod. PMID- 2549176 TI - Influence of pHo on calcium channel block by amlodipine, a charged dihydropyridine compound. Implications for location of the dihydropyridine receptor. AB - We have investigated the modulation of L-type calcium channel currents in isolated ventricular cells by the dihydropyridine derivative amlodipine, a weak base with a pKa of 8.6. Under conditions that favor neutral drug molecules, amlodipine block resembles other, previously described, neutral dihydropyridine derivatives: block is more pronounced at depolarized voltages, repetitive pulsing is not needed to promote block, and recovery is complete at hyperpolarized voltages. When the drug is ionized, depolarized voltages still enhance block, however, the time course is slow and speeded by repetitive pulses that open channels. Recovery from block by ionized drug molecules is very slow and incomplete, but can be rapidly modified by changes in external hydrogen ion concentration. We conclude from these observations that the degree of ionization of the drug molecule can affect access to the dihydropyridine receptor and that external protons can interact with the drug-receptor complex even if channels are blocked and closed. These observations place limitations on the location of this receptor in the ventricular cell membrane. PMID- 2549177 TI - Sodium-calcium exchange current. Dependence on internal Ca and Na and competitive binding of external Na and Ca. AB - Na-Ca exchange current was measured at various concentrations of internal Na [( Na]i) and Ca [( Ca]i) using intracellular perfusion technique and whole-cell voltage clamp in single cardiac ventricular cells of guinea pig. Internal Ca has an activating effect on Nai-Cao exchange beginning at approximately 10 nM and saturating at approximately 50 nM with a half maximum [Ca]i (Km[Ca]i) of 22 nM (Hill coefficient, 3.7). Measurement of Nai-Cao exchange current at various concentration of [Na]i revealed an apparent Km[Na]i of 20.7 +/- 6.9 mM (n = 14) with imax of 3.5 +/- 1.2 microA/microF. For [Ca]i transported by the exchange, a Km[Ca]i of 0.60 +/- 0.24 microM (n = 8) with an imax of 3.0 +/- 0.54 microA/microF was obtained by measuring Nao-Cai exchange current. These values are apparently different from the values for the external binding site which have been reported previously. Whether Na and Ca compete for the external binding site, and if so, how it affects the binding constants was then investigated. Outward Nai-Cao exchange current became larger by reducing [Na]o. The double reciprocal plot of the current magnitude and [Ca]o at different [Na]o revealed a competitive interaction between Na and Ca. In the absence of competitor [Na]o, an apparent Km[Ca]o of 0.14 mM was obtained. When comparing internal and external Km values, the external value is markedly larger than the internal one and thus we conclude that binding sites of the Na-Ca exchange molecule are at least apparently asymmetrical between the inside and outside of the membrane. PMID- 2549178 TI - Bovine leukaemia virus packaging cell line for retrovirus-mediated gene transfer. AB - Retroviral packaging cell lines were constructed by using the gag-pol gene of spleen necrosis virus, the gag-pol gene of Moloney murine leukaemia virus and the env gene of bovine leukaemia virus. The plasmids containing the gag-pol genes and the plasmid containing the env gene were cotransfected into NIH/3T3 and D17 cells. The cells containing the helper virus constructs were tested for their ability to package replication-defective murine leukaemia and avian reticuloendotheliosis retrovirus vectors. The titre of vector virus produced by each of the retroviral packaging cell lines was about 10(2) colony-forming units per ml of medium. Tests for events that might result in intact replication competent retroviruses showed no evidence for the generation of such viruses. The vector viruses were able to infect dog and rat cells. Bovine cells were infected only after their cocultivation with the retroviral packaging cell lines producing murine leukaemia virus vectors, perhaps as a result of a low concentration of receptors. PMID- 2549179 TI - A transcriptional map of visna virus: definition of the second intron structure suggests a rev-like gene product. AB - Visna virus is the prototype lentivirus, with a genome structure similar to that of the human immunodeficiency viruses HIV-1 and HIV-2. We have analysed in vitro the transcription pattern of this virus in lytic infections of choroid plexus cells. Northern blot analysis shows the presence of spliced subgenomic mRNA species of 4.9, 4.3, 4.0, 1.7 and 1.4 kb. Use of appropriate subgenomic probes shows that the first three of these species encode envelope protein (but also potentially the small open reading frames Q and S). The 1.7 kb RNA could contain S. In order to elucidate the translational coding potential of the smallest RNA, and to characterize further all the transcripts, S1 mapping was performed across those parts of the genome which were close to exon/intron boundaries. This allowed the definition of acceptor splice sites following both introns 1 and 2 as well as donor sites preceding intron 2. The data suggest that the 1.4 kb RNA encodes a protein derived from the F reading frame that may form part of a precursor protein and also contains sequences with some degree of homology to the rev (trs/art) protein of HIV, as well as a typical protease cleavage site between these sequences and the F protein. PMID- 2549181 TI - Molecular characterization of a neutralizing domain of the Japanese encephalitis virus structural glycoprotein. AB - Expression of antigenic fragments of the Japanese encephalitis virus envelope protein (E) in Escherichia coli has been used to define the boundaries of an antigenic domain that contains the binding sites for 10 anti-E monoclonal antibodies (MAbs). All of these antibodies neutralized the virus in vitro and some of them passively protected mice from a fatal virus challenge. We have shown previously that nine of these antibodies react with the antigenic determinants encoded by a 405 bp fragment of viral cDNA. To determine the amino acid sequences of specific determinants, truncated polypeptides were expressed as fusion proteins in E. coli following progressive Bal 31 exonuclease digestion of the 5' and 3' ends of the cDNA fragment. Examination of the immunoreactivity of these polypeptides revealed that the region from methionine 303 to tryptophan 396 was the shortest sequence capable of reacting with any of the 10 MAbs or with a polyclonal, antiviral hyperimmune mouse ascitic fluid. Biochemical tests showed that an intramolecular disulphide cross-linkage between cysteine 304 and cysteine 335 of the E protein sequence was required for presentation of the binding site(s) for these MAbs. Although this 95 amino acid antigenic domain appeared to be capable of forming several conformational neutralizing epitopes, it was not an effective immunogen for inducing neutralizing or protective antibodies in mice. PMID- 2549180 TI - The characterization of equine encephalosis virus and the development of genomic probes. AB - Equine encephalosis virus (EEV) is an orbivirus associated with a peracute illness of horses in southern Africa. The virus has now been partially purified for the first time and characterized on a molecular level. The virion is composed of 10 dsRNA segments and a protein capsid consisting of at least seven structural proteins that vary in Mr from 36,000 to 120,000. Partial clones of six of the dsRNA segments of EEV serotype Cascara were obtained and analysed for possible use as serotype-specific or group-specific probes in the detection of EEV dsRNA. Cloned fragments of genome segments 3, 8 and 10 were found to show high conservation of these segments, hybridizing to dsRNA from the six EEV serotypes under conditions that indicated more than 90% sequence homology. The genome segment 2-specific probe did not hybridize with dsRNA from any of the other EEV serotypes, suggesting that this segment encodes the serotype-specific antigen of EEV. Cross-hybridization of probes from genome segments 3 and 5 with dsRNA from bluetongue virus (BTV), epizootic haemorrhagic disease virus (EHDV) and African horse sickness virus (AHSV) indicated that EEV is more closely related to BTV and EHDV than to AHSV. Both probes can be used to distinguish between EEV and AHSV dsRNA. PMID- 2549182 TI - Replication kinetics and cytopathic effect of hepatitis A virus. AB - The replication kinetics and c.p.e. of hepatitis A virus (HAV) strain HM-175 were shown to depend upon the passage level of the cell line, and the passage level and method of selection of the virus population. Maximum virus production under single-step growth curve conditions occurred as early as 24 to 28 h or as late as 10 days post-infection. Although rapid replication of an isolate of HM-715 (pHM 175) occurred initially in BS-C-1 cells, its most pronounced c.p.e. was induced in FRhK-4 cells. The replication kinetics of pHM-175 in BS-C-1 cells were similar to those in FRhK-4 cells, although a higher yield of virus was obtained in the latter. The HAV that generated c.p.e. in FRhK-4 cells was obtained by two different selection processes: virus passage, or cloning of large focus-forming variants from the radioimmunofocus assay. The c.p.e. and yield of infectious pHM 175 in FRhK-4 cells could be reduced by 3 mM-guanidine. Another HAV isolate, strain MD-1, isolated directly from contaminated ground water in cell culture demonstrated c.p.e. in FRhK-4 cells after passage as persistently infected A-549 cells. PMID- 2549183 TI - A herpes simplex virus type 2 variant in which a deletion across the L-S junction is replaced by single or multiple reiterations of extraneous DNA. AB - We have isolated and characterized a novel variant of herpes simplex virus type 2 (HSV-2) strain HG52 which has a deletion of 13.5 kb across the L-S junction of the genome, resulting in all of the IRL region, half of the IRS region and the intervening L-S junction 'a' sequence(s) being removed. The deleted DNA has been replaced by single or multiple (up to 14) reiterations of a DNA sequence approximately 1 kb in length. Individual genomes within the population range in size from approximately 12 kb smaller than unit length to unit length or marginally larger. The L component of the genome is fixed in the prototype orientation while the S component inverts inefficiently. The variant is viable in tissue culture, is not temperature-sensitive, demonstrates impaired single-cycle growth characteristics and, apart from altered mobility of a single species (29.5K), its polypeptide profile in infected cells appears normal. Southern blot analysis has failed to identify the inserted sequences as being derived from HSV 2. PMID- 2549184 TI - A role for herpes simplex virus type 1 glycoprotein E in induction of cell fusion. AB - The role of herpes simplex virus type 1 (HSV-1) glycoprotein E (gE) in the induction of multinucleate cell (syncytium) formation was investigated using monoclonal antibodies and a gE deletion mutant, R7023. We found that monoclonal antibodies directed against gE blocked HSV-1-induced syncytium formation in human cells. R7023 also failed to induce syncytium formation in tissue culture cells. The results indicate that gE, in addition to glycoproteins B, D, H and the gene sequence located between 0.732 and 0.745 map units, is involved in cell fusion. Thus, this important biological property appears to be regulated by several HSV-1 gene products. PMID- 2549185 TI - In vitro promoter activity associated with the latency-associated transcript gene of herpes simplex virus type 1. AB - The herpes simplex virus latency-associated transcript (LAT) gene is the only viral gene that shows substantial transcriptional activity during neuronal latency. The LAT RNA produced is antisense to the mRNA of the immediate early gene ICP0, partially overlaps the ICP0 mRNA, and is suspected of playing some role in latency. Sequence analysis of the region 5' to the reported transcription start site has not revealed any high consensus RNA polymerase II promoter elements. Nonetheless, LAT RNA is transcribed in low abundance during acute infection in tissue culture. As the initial step in mapping the promoter for this latency-associated gene, we analysed the ability of different regions of the LAT gene to drive the transcription of an indicator gene in vitro. Using chloramphenicol acetyltransferase (CAT) assays, we found that the genomic region between -940 and -662 nucleotides upstream of the transcription start site of the LAT gene was most efficient at directing transcription of the indicator CAT gene in Vero cells. This suggests that the LAT promoter, or at least the promoter controlling transcription of this gene during acute infection in tissue culture, may have an unusual location of more than 662 nucleotides upstream from the reported start of RNA transcription. PMID- 2549186 TI - Retention and expression of the left end subfragment of the herpes simplex virus type 2 BglII N DNA fragment do not correlate with tumorigenic conversion of NIH 3T3 cells. AB - Cotransfection experiments have been carried out using recombinant plasmids pAG60, conferring resistance to antibiotic G418, and pXho3 which contains the left end subfragment (map coordinates 0.583 to 0.596) of the transforming herpes simplex virus type 2 BglII N DNA fragment and encodes the 36K polypeptide associated with the viral ribonucleotide reductase activity. Several NIH 3T3 cell clones resistant to G418 and having morphological changes commonly observed for transformed NIH 3T3 cells were isolated and examined for the presence and stable retention of the viral sequences. Seven of the clones that retained the transfected viral sequences were analysed for the expression of the 36K polypeptide and the tumorigenic phenotype. The results gathered from these studies show that neither the retention of the viral DNA nor the expression of the 36K polypeptide correlated with tumorigenic conversion of these cells. PMID- 2549187 TI - Beta 2 microglobulin binds to the tegument of cytomegalovirus: an immunogold study. AB - Previous reports have provided evidence for the ability of human cytomegalovirus (HCMV) to bind the host protein beta 2 microglobulin (beta 2m) from body fluids or culture medium, and thus enhance infectivity of the virus, both by evasion of immune neutralization and the capacity to employ the bound beta 2 m for attachment to the host cell. Immunocytochemical techniques and negative stain electron microscopy were used to identify the ultrastructural components of HCMV involved in its interaction with beta 2m. Probes comprising colloidal gold coupled to beta 2m were seen to bind not to the envelope as previously suspected, but to material closely surrounding the nucleocapsids. It is postulated that the tegument proteins of HCMV, via their capacity to bind beta 2m, play an important role in the preservation of infectivity of disrupted virions by enabling unenveloped capsids to bind to cells and gain entry by a pathway other than that normally taken by intact virions. PMID- 2549188 TI - Carboxy-terminal analysis of nine proteins specified by the flavivirus Kunjin: evidence that only the intracellular core protein is truncated. AB - Nine proteins specified by Kunjin virus were labelled with [3H]lysine and digested with carboxypeptidase B which specifically cleaves carboxy-terminal Lys or Arg. The theoretical amount of [3H]lysine was released from the non-structural (ns) proteins NS2A, NS2B, NS3 and NS4B, which have a common carboxy terminus Lys Arg deduced from cleavage sites established in the viral polyprotein by previous N-terminal amino acid analyses. This is a flavivirus consensus site, always followed by Gly, Ala or Ser. These results indicate that no truncations had occurred despite anomalous electrophoretic migrations of NS2A, NS2B and NS4B observed in some gel systems. No [3H]lysine was released from NS4A or NS5 which terminate in Ala and Leu, respectively, thus establishing that NS5 (observed Mr 98,000, theoretical Mr 103,600) was not cleaved post-translationally at any internal Lys-Arg site. Unexpectedly, [3H]lysine residues were apparently released from P14(C), the intracellular equivalent of virion C protein which terminates in Ala (adjacent to the established N-terminus of prM). However, a putative internal cleavage site (Lys-Arg decreases Gly) exists 18 residues upstream from the carboxy terminus of C, prior to the transmembrane spanning domain. Apparent internal cleavage in the cytosol at this site to produce P14(C) would expose [3H]lysine residues to carboxypeptidase B, and account for the previously observed differences in size and composition between C and P14(C). PMID- 2549189 TI - Modulation of lentivirus replication by antibodies. Non-neutralizing antibodies to caprine arthritis-encephalitis virus enhance early stages of infection in macrophages, but do not cause increased production of virions. AB - Non-neutralizing antibodies to caprine arthritis-encephalitis virus (CAEV) enhance the early stages of the virus life cycle but do not potentiate enhanced production of virus particles by macrophages. In primary macrophages used for these studies, there was enhancement in binding, internalization and uncoating of virus pretreated with non-neutralizing sera in comparison to virus pretreated with a non-immune serum. However, this did not lead to enhanced production of virus particles. Failure of non-neutralizing sera to inactivate CAEV may be due in part to low avidity of the antibodies for the virus particles which contain sialic acids on their envelopes, because desialylation of the particles made them neutralizable. The non-neutralizing antibodies probably bound to most of the native virus particles which were then internalized via Fc receptor-mediated endocytosis and degraded. Sialylated particles that failed to bind antibodies probably caused the infection. Thus there was no true enhancement of infection. The previously reported increase in severity of lesions in animals immunized with inactivated CAEV particles prior to challenge with live virus suggested enhancement of infection but in the light of our finding this may have been caused by factors other than an increase in production in the number of infectious virus particles. PMID- 2549190 TI - Oestrogen stimulates differential transcription of human papillomavirus type 16 in SiHa cervical carcinoma cells. AB - Human papillomavirus (HPV) type 16 is highly associated with cervical cancer, but it seems that cofactors such as hormones affect its potential oncogenicity. We have analysed the HPV-16 gene expression in response to sex hormones and glucocorticoids in SiHa cells, a human cervical carcinoma cell line. An eightfold induction of HPV-16 transcripts was obtained in oestrogen-treated SiHa cells. Of the five HPV-16 transcripts detected in these cells only the two major ones, the 4.6 kb and the 4.1 kb mRNA species, were affected by oestrogen. Since the five transcripts span the E6 and E7 open reading frames of the HPV-16 genome, these results suggest that the expression of the various transcripts is differentially controlled, as oestrogen regulates only two of them. We have identified in the HPV-16 genome seven different regions with a high degree of similarity to the oestrogen-responsive element consensus sequence (GGTCANNNTGACC). These sequences are located throughout the entire HPV-16 genome. Progesterone or dexamethasone had no detectable stimulatory effect on the various transcripts of HPV-16 in SiHa cells, up to 24 h after treatment of the cells. Since the E6 and E7 open reading frames have been associated with the oncogenic potential of HPV-16, the effect of oestrogen on the transcription of these viral genes may be of biological relevance in the malignant transformation of HPV-16-infected cervical cells. PMID- 2549191 TI - Antidepressant treatments, including sibutramine hydrochloride and electroconvulsive shock, decrease beta 1- but not beta 2-adrenoceptors in rat cortex. AB - The beta 1- and beta 2-adrenoceptor populations in rat cortex were individually quantified by labelling all of the receptors with [3H]dihydroalprenolol and displacing with isoprenaline (200 microM) or CGP 20712A (1-(2-[(3-carbamoyl-4 hydroxy)phenoxy]ethylamino)-3-[4-(1-methyl-4- trifluoromethyl-2 imidazolyl)phenoxy]-2-propanol methanesulphonate; 100 nM) to define total beta adrenoceptors and beta 1-adrenoceptors, respectively. Binding parameters for beta 2-adrenoceptors were calculated by the difference. Oral administration of the monoamine reuptake inhibitors sibutramine HCl (3 mg/kg), amitriptyline (10 mg/kg), desipramine (10 mg/kg), or zimeldine (10 mg/kg) for 10 days decreased the total number of beta-adrenoceptors present in rat cortex. This effect was entirely due to a reduction in the number of beta 1-adrenoceptors. Similarly, 10 days of treatment with the monoamine oxidase inhibitor tranylcypromine (10 mg/kg p.o.) or five electroconvulsive shocks (ECSs; 200 V, 2 s) spread over this period also down-regulated beta-adrenoceptors by reducing the content of the beta 1 subtype. By contrast, treatment with clenbuterol (5 mg/kg p.o.) for 10 days reduced the number of cortical beta-adrenoceptors by an effect on the beta 2 adrenoceptor population. The effects of short-term treatment with these drugs were also investigated, and, using the doses shown above, the results of 3 days of administration or a single ECS were determined. Sibutramine HCl and desipramine were alone in producing a reduction in number of beta-adrenoceptors after 3 days. Once again, this was exclusively due to a loss of beta 1 adrenoceptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549192 TI - Characterization of insulin-like growth factor-I receptors in PC12 pheochromocytoma cells and bovine adrenal medulla. AB - Competitive binding studies indicated that PC12 cells have receptors for insulin like growth factor-I (IGF-I). There are approximately 11,000 +/- 1,500 IGF-I receptors/cell; these receptors have an apparent KD for IGF-I of 7.2 +/- 0.6 nM. Covalent cross-linking of 125I-IGF-I to PC12 cells labeled a 125,000-130,000-Mr protein, presumably the alpha-subunit of the IGF-I receptor. Although PC12 cells also have insulin receptors, the 125I-IGF-I appeared to be cross-linked to IGF-I receptors, because 100 nM IGF-I competed for labeling but 100 nM insulin did not. Bovine chromaffin cells also have IGF-I receptors. The protein tyrosyl kinase activity of IGF-I receptors from bovine adrenal medulla and PC12 cells was examined after purification of the receptors by wheat germ agglutinin-Sepharose chromatography. IGF-I (10 nM) stimulated autophosphorylation of the beta-subunits of the IGF-I receptors from both preparations; the beta-subunits from both sources had Mr values of approximately 97,000. IGF-I also stimulated phosphorylation of the synthetic substrate poly(Glu:Tyr)4:1 by both receptor preparations. IGF-I (IC50 of approximately 0.2 nM) was much more potent than insulin at stimulating phosphorylation of poly(Glu:Tyr) by the bovine adrenal medulla preparation. A maximal concentration of IGF-I (10 nM) increased phosphorylation approximately threefold. IGF-I was slightly more effective than insulin at stimulating the phosphorylation of poly(Glu:Tyr) by the PC12 cell receptor preparation, but neither ligand produced a maximal effect at concentrations up to 100 nM. This result probably reflects the presence of comparable numbers of IGF-I and insulin receptors on PC12 cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549193 TI - Separate binding and functional sites for omega-conotoxin and nitrendipine suggest two types of calcium channels in bovine chromaffin cells. AB - Purified adrenomedullary plasma membranes contain two high-affinity binding sites for 125I-omega-conotoxin, with KD values of 7.4 and 364 pM and Bmax values of 237 and 1,222 fmol/mg of protein, respectively. Dissociation kinetics showed a biphasic component and a high stability of the toxin-receptor complex, with a t1/2 of 81.6 h for the slow dissociation component. Unlabeled omega-conotoxin inhibited the binding of the radioiodinated toxin, adjusting to a two-site model with Ki1 of 6.8 and Ki2 of 653 pM. Specific binding was not affected by Ca2+ channel blockers or activators, cholinoceptor antagonists, adrenoceptor blockers, Na+ channel activators, dopaminoceptor blockers, or Na+/H+ antiport blockers, but divalent cations (Ca2+, Sr2+, and Ba2+) inhibited the toxin binding in a concentration-dependent manner. The binding of the dihydropyridine [3H]nitrendipine defined a single specific binding site with a KD of 490 pM and a Bmax of 129 fmol/mg of protein. At 0.25 microM, omega-conotoxin was not able to block depolarization-evoked Ca2+ uptake into cultured bovine adrenal chromaffin cells depolarized with 59 mM K+ for 30 s, whereas under the same conditions, 1 microM nitrendipine inhibited uptake by approximately 60%. When cells were hyperpolarized with 1.2 mM K+ for 5 min and then Ca2+ uptake was subsequently measured during additions of 59 mM K+. Omega-conotoxin partially inhibited Ca2+ uptake in a concentration-dependent manner. These results suggest that two different types of Ca2+ channels might be present in chromaffin cells. However, the molecular identity of omega-conotoxin binding sites remains to be determined. PMID- 2549194 TI - GABAA receptor complex in an experimental model of hepatic encephalopathy: evidence for elevated levels of an endogenous benzodiazepine receptor ligand. AB - The involvement of the gamma-aminobutyric acidA (GABAA) receptor complex in the pathogenesis of hepatic encephalopathy was examined in thioacetamide-treated rats with fulminant hepatic failure. Partially purified extracts from encephalopathic rat brain were approximately three times more potent in inhibiting [3H]Ro 15-1788 binding to benzodiazepine receptors than identically prepared extracts from control rats. High levels of inhibitory activity were also found in extracts of plasma, heart, and liver from thioacetamide-treated rats. The inhibition of [3H]Ro 15-1788 binding by brain extracts appeared to be competitive and reversible and was unaffected by treatment with either proteolytic enzymes or boiling. Further, GABA significantly enhanced the potency of these extracts in inhibiting [3H]flunitrazepam binding. In contrast, no differences were found in radioligand binding to the constituent recognition sites of the GABAA receptor complex in well-washed brain membranes prepared from control and encephalopathic animals. These findings suggest that the recognition-site qualities of the constituent proteins of the GABAA receptor complex are unchanged in an experimental model of hepatic encephalopathy. However, significant elevations in the level of a substance or substances with neurochemical properties characteristic of a benzodiazepine receptor agonist may contribute to the electrophysiological and behavioral manifestations of hepatic encephalopathy. PMID- 2549195 TI - Ascorbic acid uptake by a high-affinity sodium-dependent mechanism in cultured rat astrocytes. AB - Ascorbic acid (vitamin C) is synthesized in rodent liver, circulates in the blood, and is concentrated in the brain. Experiments were performed to characterize the mechanism of ascorbate uptake by rat cerebral astrocytes in primary culture. Astroglial uptake of L-[14C]ascorbate was observed to be both saturable and stereoselective. In addition, uptake was dependent on both the incubation temperature and the concentration of Na+ because it was largely inhibited by cooling to 4 degrees C, by treatment with ouabain to increase intracellular Na+, and by the substitution of K+, Li+, or N-methyl-D-glucamine for extracellular Na+. The affinity for ascorbate was relatively high in cells incubated with a physiological concentration of extracellular Na+, because the apparent Km was 32 microM in 138 mM Na+. However, the affinity for ascorbate was significantly decreased when the extracellular Na+ concentration was lowered. Treatment of astrocytes with dibutyryl cyclic AMP induced stellation and increased the maximum rate of ascorbate uptake by 53%. We conclude that astrocytes possess a stereoselective, high-affinity, and Na+-dependent uptake system for ascorbate. This system may regulate the cerebral ascorbate concentration and consequently modulate neuronal function. PMID- 2549196 TI - Immunological identification of multiple alpha-like subunits of the gamma aminobutyric acidA receptor complex purified from neonatal rat cortex. AB - Antibodies were prepared against a synthetic peptide corresponding to amino acid sequences 174-203 of the bovine gamma-aminobutyric acidA (GABAA) receptor alpha 1 subunit. The antibodies recognized this synthetic alpha 1-peptide, but failed to react with the homologous peptide sequence, 170-199, of the bovine beta 1 subunit. On Western blots, anti-alpha 1-subunit antibody recognized a 50 kilodalton (kDa) protein in affinity-purified receptor preparations from adult rat cortex and cerebellum. In receptor purified from neonatal cortex, the anti alpha 1-antibody reacted with 50-kDa, 53-54-kDa, and 59-kDa proteins. After digestion with endoglycosidase F, these three protein bands retained differing electrophoretic mobilities. The 50-kDa and 59-kDa subunits of affinity-purified neonatal receptor, which were photoaffinity-labeled with [3H]flunitrazepam, were immunoprecipitated to different extents by alpha-subunit antibody. These data suggest the existence in GABAA receptor from neonatal cortex of three proteins (50 kDa, 53 kDa, and 59 kDa) which have immunological homology to alpha 1-subunit of bovine GABAA receptor. The presence of an alpha- and a beta-like subunit with similar mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis may account for the relatively high concentration of protein in the 53-54-kDa band which has been observed in receptor purified from neonatal cortex. The presence of multiple alpha-like subunits may be related to the presence of a relatively high concentration of type II GABA receptor in this tissue. PMID- 2549197 TI - Characterization of receptor-coupled phosphoinositide hydrolysis in the rat hippocampus after intradentate colchicine. AB - Lesions produced by intradentate hippocampal administration of colchicine have been reported to produce several time-dependent behavioral and neurochemical changes, including a possible change in the signal transduction process for the cholinergic muscarinic receptor. To characterize further the effects of colchicine on receptor-coupled hydrolysis of phosphoinositides, colchicine was injected stereotaxically into the dentate gyrus of rats at a dose of 2.5 micrograms/site. The animals were killed 1, 3, or 12 weeks after injection and the hippocampi removed and sliced. [3H]Inositol was incorporated into slices, and various receptor agonists known to stimulate inositol phosphate (IP) metabolism were studied. Colchicine administration altered agonist-stimulated turnover in the hippocampus in a time-dependent manner. This hyperstimulation was receptor mediated, because it was blocked by pirenzepine. The hyperstimulation of turnover was observed also with norepinephrine and serotonin. Colchicine had no effect on IP turnover in vitro. The effect of the colchicine lesion was observed only in the hippocampus, because no change in cholinergic muscarinic receptor-stimulated phosphatidylinositol turnover was observed in the cortex. These studies indicate that intradentate administration of colchicine produces a compensatory change in the signal transduction process in the hippocampus detectable 12 weeks after the lesion. PMID- 2549198 TI - Rat brain synaptosomal ATP:AMP-phosphotransferase activity. AB - Adenylate kinase activity (ATP:AMP-phosphotransferase; EC 2.7.4.3) was studied in various subcellular fractions of rat brain tissues. Because of the presence of other adenosine nucleotide-utilizing enzymes, adenylate kinase activity was assayed in both the forward and reverse directions by using coupled enzyme systems and by using a specific adenylate kinase inhibitor, P1,P5-di(adenosine 5') pentaphosphate. As expected, the highest specific adenylate kinase activity (2.89 mumol/min/mg of protein) was detected in the cytosolic brain fraction. However, substantial enzyme activity (0.68 mumol/min/mg) was also found in the intact synaptosomal fraction isolated on Percoll/sucrose gradients. The increased specific enzyme activity of purified synaptosomes and the differences found between the kinetic parameters of the membrane-bound and cytosolic enzyme forms suggest that the synaptosomal adenylate kinase activity cannot be attributed to the small amount of contaminating cytosol present in our preparations. The adenylate kinase enzyme adhered to purified synaptic plasma membranes and was not released by washings with isoosmotic sucrose medium. The facts that the adenylate kinase enzyme activity could be measured in intact synaptosomal preparations and that both its substrates and its inhibitors do not cross intact plasma membranes support the possibility that the synaptosomal adenylate kinase is an ecto-enzyme. PMID- 2549199 TI - Capsaicin-induced ion fluxes increase cyclic GMP but not cyclic AMP levels in rat sensory neurones in culture. AB - Capsaicin, which induces fluxes of sodium, calcium, and potassium ions in a subset of both neonatal and adult rat dorsal root ganglion neurones, increased cyclic GMP (cGMP) levels by a factor of 20 (EC50 0.07 microM) to 10-20 pmol cGMP/mg protein in these cells. Cyclic AMP (cAMP) levels were unaffected. Nonneuronal cells derived from rat ganglia, and both neurones and nonneuronal cells from chick were unresponsive to capsaicin. Capsaicin-induced cGMP elevation in rat dorsal root ganglion (DRG) neurones was unaffected by pertussis toxin, lowered by compounds that block voltage-sensitive calcium channels, and was abolished by the removal of extracellular calcium. Calcium, guanidine, and rubidium fluxes were unaffected by treatment of DRG cells with sodium nitroprusside or dibutyryl cGMP. The cGMP response to capsaicin is thus a function of capsaicin-evoked calcium uptake through voltage-sensitive calcium channels. Elevated cGMP levels do not, however, contribute to capsaicin-evoked ion fluxes or to their desensitisation. PMID- 2549200 TI - Mixed competitive and allosteric antagonism by gallamine of muscarinic receptor mediated second messenger responses in N1E-115 neuroblastoma cells. AB - The antagonistic effects of gallamine on muscarinic receptor-linked responses were investigated in N1E-115 neuroblastoma cells. M1 muscarinic receptor-mediated phosphoinositide hydrolysis induced by carbamylcholine was antagonized by gallamine, with a Ki value of 33 microM. By comparison, gallamine was four- to fivefold less potent in blocking noncardiac M2 muscarinic receptor-mediated inhibition of cyclic AMP formation, with a Ki value of 144 microM. The resulting Arunlakshana-Schild plots of the antagonism of both responses by gallamine were linear and exhibited slopes not differing from 1, a result indicative of a competitive mechanism. To elucidate further the nature of gallamine's inhibitory actions, experiments were performed where the effects of gallamine in combination with the known competitive muscarinic antagonist, N-methylscopolamine (NMS), were studied. In the presence of both antagonists, a supraadditive shift in the carbamylcholine dose-response curve was demonstrated for the two responses, a result suggestive of an allosteric mode of interaction between gallamine and NMS binding sites. Confirmation that gallamine allosterically modifies the muscarinic receptor was provided by radioligand binding studies. Gallamine competition curves with either [N-methyl-3H]scopolamine methyl chloride ([3H]NMS) or [N methyl-3H]quinuclidinyl benzilate methyl chloride ([3H]NMeQNB) were unusually shallow. Furthermore, gallamine decelerated the rate of dissociation of receptor bound [3H]NMS greater than [3H]NMeQNB in a dose-dependent manner. The present study demonstrates that whereas gallamine antagonizes carbamylcholine-mediated responses in N1E-115 cells in a competitive manner, an allosteric component of its action is revealed in the presence of muscarinic antagonists such as NMS. PMID- 2549201 TI - Diphtheritic neuropathy, an analysis based on muscle and nerve biopsy and repeated neurophysiological and autonomic function tests. AB - A patient with diphtheritic neuropathy was investigated with repeated tests of parasympathetic and sympathetic vasomotor and sudomotor functions for one year after the onset of symptoms. Somatic nerve function was tested with nerve conduction studies and an index based on ten variables was used to follow the course of the neuropathy. Sural nerve and anterior tibial muscle biopsies were performed. A severe but shortlasting impairment of the parasympathetic vagal reflex arc was found. The recovery of this function paralleled the clinical course. Sympathetic functions were normal. The neurophysiological variables of somatic nerve function showed signs of a mainly demyelinating mixed sensory/motor neuropathy. The recovery of these variables was slow. The nerve and muscle biopsies demonstrated mild changes consistent with a mixed, demyelinating, non inflammatory neuropathy. PMID- 2549202 TI - Alpha 2-adrenoceptor-mediated modulation of noradrenaline release is decreased in vas deferens but not in cerebral cortex of diabetic rats. AB - In the present study, a superfusion method was used to investigate the alpha 2 adrenoceptor mediated modulation of the electrically evoked release of radiolabeled noradrenaline in vas deferens and cerebral cortex slices of streptozotocin-diabetic and control rats 8-10 weeks after the induction of diabetes. Stimulation of alpha 2-adrenoceptors led to a significantly smaller inhibition of radiolabeled noradrenaline release in the vas deferens of diabetic rats as compared to control rats. In the cerebral cortex no such difference was detected. It is concluded that these effects could be due to a decrease in the number of presynaptic alpha 2-adrenoceptors in the vas deferens of diabetic rats. The results of this study show that the superfusion technique offers a simple possibility to obtain information about the functional integrity of noradrenergic neurons in the peripheral nervous system and central nervous system in diabetes mellitus. Moreover, the results provide an indication for a different time scale in the development of neuropathy in the peripheral and central nervous system of streptozotocin-diabetic rats. PMID- 2549203 TI - Pathological prognostic factors in stage I (T1N0M0) and stage II (T1N1M0) breast carcinoma: a study of 644 patients with median follow-up of 18 years. AB - Prognostic factors have been examined in 644 patients with tumor-node-metastasis (TNM) stage T1 breast carcinoma treated by mastectomy and followed for a median of 18.2 years. Overall, 148 patients (23%) died of recurrent breast carcinoma. Eighteen (3%) were alive with recurrent disease and 478 (74%) were alive or died of other causes without recurrence. Unfavorable clinicopathologic features were larger tumor size (1.1 to 2.0 cm v less than or equal to 1 cm), perimenopausal menstrual status, the number of axillary lymph node metastases, poorly differentiated grade, presence of lymphatic tumor emboli (LI) in breast tissue near the primary tumor, blood vessel invasion (BVI), and an intense lymphoplasmacytic reaction around the tumor. Median survival after recurrence for the entire series was 2 years. This was not significantly influenced by tumor size, the number of axillary nodal metastases, the type of treatment for recurrence, or the interval to recurrence. The proportions surviving 5 and 10 years after recurrence were 17% and 5%, respectively. Among T1N0M0 cases, the chance of a local recurrence was 2.8% within 20 years. Median survival of T1N0M0 cases after local recurrence (4.5 years) was significantly longer than after systemic recurrence (1.5 years). A similar trend (3.7 v 2.0 years), not statistically significant, was seen in T1N1M0 patients, who had a 6.5% chance of local recurrence within 20 years. Median survival following systemic recurrence detected 10 or more years after diagnosis in T1N0M0 and in T1N1M0 patients was significantly longer than the median survival for systemic recurrences found in the first decade of follow-up. This difference did not apply following local recurrence in either T1N0M0 or T1N1M0 cases. It is evident that patients with T1 breast carcinoma can be subdivided into differing prognostic groups and this must be taken into account when considering the role of adjuvant chemotherapy for stage I disease. Systemic adjuvant treatment may prove to be beneficial for patients with unfavorable prognostic factors, while women with an especially low risk for recurrence (eg, T1N0M0 tumor 1.0 cm or less) might be spared such treatment. PMID- 2549204 TI - A randomized trial to evaluate the effect of schedule on the activity of etoposide in small-cell lung cancer. AB - Etoposide is an increasingly used and well-tolerated drug in cancer medicine. Its cytotoxic action is phase-specific and it has demonstrated schedule dependency in both in vitro and animal studies, but clinical evidence of the importance of drug scheduling is uncertain. The two administration schedules of etoposide that have been compared in this randomized study of 39 patients with previously untreated extensive small-cell lung cancer treated with single-agent etoposide were 500 mg/m2 as a continuous intravenous (IV) infusion over 24 hours or five consecutive daily 2-hour infusions each of 100 mg/m2. Both regimens were repeated every 3 weeks, for a maximum of six cycles. Patients received combination chemotherapy with vincristine, doxorubicin, and cyclophosphamide (VAC) or radiotherapy on failure to respond or at relapse, depending on their Karnofsky performance status. The same therapy was used in both arms of the study. All patients are evaluable for response to etoposide. In the 24-hour arm, two patients achieved a partial remission, resulting in an overall response rate of 10%. In the 5-day schedule, 16 patients had a partial response and one had a complete remission, producing an overall response rate of 89%, which was significantly superior to that in the 24-hour arm (P less than .001). The median duration of remission to etoposide in the 5-day arm was 4.5 months. Bone marrow toxicity was similar in both schedules. Etoposide pharmacokinetics were measured in all patients, and total areas under the concentration versus time curves (AUCs) were equivalent in both regimens. This study has clearly demonstrated the importance of etoposide scheduling in humans, and the superiority of five daily infusions over a 24-hour continuous infusion. The response rate to single-agent etoposide using an efficacious schedule in extensive small-cell lung cancer has been determined to be in excess of 80%. PMID- 2549205 TI - Phase I clinical investigation of amonafide. AB - Amonafide (benzisoquinolinedione, NSC 308847) is a new synthetic imide antineoplastic agent with DNA intercalative properties that has been evaluated in a phase I clinical trial. The drug was administered as a single intravenous (IV) infusion over 30 to 120 minutes repeated every 28 days. Ninety-five courses of therapy at doses ranging from 18 to 1,104 mg/m2 were administered to 38 patients with refractory solid tumors. Granulocytopenia was dose limiting. Leukopenia was seen in 13 of 31 courses at doses of 690 mg/m2 or greater. Life-threatening granulocytopenia (less than or equal to 250 microliters) was noted in 1/6 patients treated at 800 mg/m2, 1/8 patients treated at 918 mg/m2, and 2/5 patients treated at 1,104 mg/m2. No definite relationship between myelotoxicity and prior treatment status was noted. Rate-of-infusion dependent, nonhematologic toxicities included diaphoresis, flushing, dizziness, and tinnitus, all of which were ameliorated by increasing the duration of drug infusion to 120 minutes. In addition, nausea and vomiting (grades 1 and 2) were seen in 29/56 courses at doses greater than or equal to 519 mg/m2, but were easily controlled by phenothiazine antiemetics. Amonafide plasma and urine concentrations were determined by high-pressure liquid chromatography (HPLC). Plasma concentrations declined biexponetially with a terminal harmonic mean terminal half-life (t 1/2) of 5.5 h. The mean apparent volume of distribution at steady-state and total body clearance were 532 L/m2 and 84 L/h/m2, respectively. Less than 5% of the total dose of amonafide was excreted unchanged in the urine. Antitumor activity has been noted in one patient with non-small-cell lung cancer (one complete response exceeding 29 months duration) and in one patient with prostatic cancer (complete pain relief and improvement in bone scan for 9 months). The recommended dose for phase II trials with this schedule of amonafide is 918 mg/m2 with dose escalation to amonafide is 918 mg/m2 with dose escalation to myelotoxicity. PMID- 2549206 TI - Effects of preventing reinnervation on axotomized spinal motoneurons in the cat. II. Changes in group Ia synaptic function. AB - 1. Composite excitatory postsynaptic potentials (EPSPs) evoked by electrical stimulation of heteronymous group Ia afferents have been studied at various postoperative times in axotomized motoneurons that were denied the opportunity to reinnervate muscle. 2. The medial gastrocnemius (MG) nerve was transected and sutured onto the surface of the normally innervated lateral gastrocnemius (LG) muscle. The denervated MG muscle was excised thereby eliminating access of regenerating MG motor axons to vacant end-plates. 3. The mean amplitude of monosynaptic Ia EPSPs evoked by electrical stimulation of the LG-soleus (LGS) nerve and recorded in axotomized MG motoneurons showed an initial decline at 20 days postoperative (DPO) that was not significant. At 44 DPO, mean amplitude had declined significantly to 43% of the control mean amplitude. At 90 DPO, mean EPSP amplitude was not significantly different from control. At the latest postoperative time (150-180 DPO), mean amplitude was significantly less than the control amplitude. 4. Mean EPSP rise time (time-to-peak) was significantly increased (27%) at the earliest postoperative times (20-44 DPO). At later postoperative times (90-180), mean EPSP rise time was not significantly different from mean control rise time. 5. "Partial responses" superimposed on EPSPs were not observed at any postoperative time. 6. Mean posttetanic potentiation (PTP) of the LGS EPSP was significantly depressed at 20 DPO. At later postoperative times, PTP did not differ significantly from mean control PTP. 7. The possibility is considered that postaxotomy alterations in the electrical properties of motoneurons may explain these complex variations of mean EPSP amplitude and rise time. PMID- 2549207 TI - Excitatory neurotransmission activates voltage-dependent properties in neurons in spinal motor system of lamprey. AB - 1. Ventral horn neurons were studied under voltage clamp during episodes of sensory-evoked rhythmic coactivation in the in vitro lamprey spinal cord-tailfin preparation in the presence of strychnine. 2. Voltage clamp under a range of holding potentials during episodes of rhythmic coactivation revealed inward currents coincident with ventral root bursting in the same hemisegment and an apparent reversal potential of about -10 mV. 3. The current-voltage relationship of the peak inward current during each burst of this activity demonstrated a marked voltage dependency. 4. The voltage dependence of the inward current was eliminated by the specific NMDA-receptor blocker, APV, and by removal of Mg2+ from the bathing solution. 5. At depolarized potentials a long-lasting outward current could be observed, indicating an apparent voltage-dependent conductance for K+ and/or Cl-. This current was also blocked by APV and increased by the removal of Mg2+. 6. These results provide evidence that during rhythmic coactivation in strychnine, endogenous release of excitatory amino acid transmitter induces nonlinear conductance properties in ventral horn neurons by the activation of NMDA receptors. The results provide additional evidence that such activation contributes to the membrane potential oscillations that underlie rhythmic locomotory activity. PMID- 2549208 TI - Vagal afferent modulation of spinal nociceptive transmission in the rat. AB - 1. The effects of vagal afferent stimulation (VAS) on spinal nociceptive transmission and the spinal pathway(s) mediating VAS-produced effects were examined in pentobarbital sodium- and urethane-anesthetized, paralyzed rats. The 60 units studied responded to mechanical stimuli and noxious heating (50 degrees C) of cutaneous receptive fields confined to the glabrous skin of the toes and footpads. Recording sites were located in laminae I-VI of the L3-L5 spinal segments. 2. VAS facilitated and inhibited neuronal responses to heat. In pentobarbital-anesthetized rats, responses of most (24/44) units were facilitated by low and inhibited by higher intensities of VAS. Responses of some units (15/44) were only inhibited and others (4/44) only facilitated by VAS. Inhibition produced by VAS was intensity-, pulse width-, frequency-, and stimulation duration-dependent. In urethane-anesthetized rats, responses of 6/16 units were initially facilitated, then inhibited as the intensity of VAS was increased; responses of nine units were inhibited by VAS. Quantitative comparisons of recruitment indices, mean thresholds for inhibition and mean intensities to inhibit unit responses to heat to 50% of control revealed no significant differences between the two anesthetic conditions. 3. The effects of VAS on neuronal responses to heat were dissociable from its effect on blood pressure. Regardless of the effect of VAS on unit responses to noxious heat, VAS consistently produced intensity-dependent depressor responses. The latencies to onset of inhibition and facilitation by VAS were determined by a cumulative sum technique and bin-by-bin analysis of peristimulus time histograms. The apparent latencies were 91 +/- 11 (SE) ms for inhibition and 278 +/- 59 ms for facilitation, both of which occurred before changes in blood pressure. Finally, microinjections of lidocaine into the ventrolateral funiculus (VLF) or transections of the dorsolateral funiculus (DLF) of the thoracic spinal cord attenuated VAS-produced effects on neuronal responses, but did not affect VAS induced depressor responses. 4. The responses of 11 dorsal horn units to graded noxious heating of the skin were studied; the stimulus-response functions (SRF) were linear and monotonic throughout the temperature range examined (42-52 degrees C). VAS at intensities which inhibited unit responses to heat significantly decreased the slope of the SRF. VAS at intensities which facilitated unit responses to heat produced a leftward, parallel shift of the SRF.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2549209 TI - Analysis of vertebrate eye movements following intravitreal drug injections. III. Spontaneous nystagmus is modulated by the GABAa receptor. AB - 1. Turtle eye movements were recorded in response to horizontal motion of patterned stimuli and intravitreal injections of selective GABAergic drugs by using a contact lens search-coil technique. Similar to results from rabbit and cat, injection of picrotoxin into the turtle's eye results in a spontaneous horizontal nystagmus, with its slow-phase movement in a temporal-to-nasal direction with respect to the injected eye. In contrast, there were no prominent vertical eye movements in response to either horizontal optokinetic stimuli or drug injections. 2. Injections of bicuculline or bicuculline methyl iodide (BMI), which selectively block the GABAa receptor, had effects similar to those of picrotoxin. The GABAa agonist muscimol, on the other hand, blocked optokinetic nystagmus (OKN). Furthermore, combinations of these drugs demonstrated competitive interactions between the agonists and antagonists. 3. The threshold dose for the eye-movement effects of each drug was ascertained with the use of a radioactive calibration procedure. Tritiated inulin was injected into the vitreous. After 1 h, ocular components were assayed for radioactivity. Then, by the use of an estimate of vitreal/retinal dilution, the retinal concentrations of these threshold doses were calculated. The computed threshold retinal concentrations of the GABAa drugs were found to be in the low micromolar range. 4. These results are discussed in terms of the directionally sensitive (DS) processing which occurs in the retina, and the output of retinal DS cells to pathways involved in oculomotor control of retinal image stabilization. It is known that intravitreal application of picrotoxin makes DS retinal ganglion cells lose their selectivity for any one direction. Based on the effect of picrotoxin on OKN, it is argued that DS retinal cells provide a major input to oculomotor subsystems involved in the stabilization of gaze. Furthermore, these intravitreal drug effects on OKN are selective for GABAa drugs, suggesting that GABAa receptors play a major role in DS processing in the retina. PMID- 2549210 TI - Kainate receptor-mediated synaptic currents in mudpuppy inner retinal neurons reduced by D-O-phosphoserine. AB - 1. The effects of D-O-phosphoserine (DOS) were examined on proximal neurons in the superfused mudpuppy retinal-eyecup preparation by measuring their synaptically evoked whole-cell currents with the use of patch-clamp electrodes. 2. DOS reduced the light-evoked excitatory postsynaptic potentials (EPSPs) of amacrine and ganglion cells. This suppression was present even though the center responses of both ON- and OFF-bipolar cells were unaffected by DOS. 3. When recordings were done under voltage-clamp conditions. DOS diminished the magnitude of light-evoked synaptic currents associated with a reduction in synaptic conductance. 4. To determine which acidic amino acid receptor mediated the network-selective action of DOS, various glutamate agonists were tested against this excitatory amino acid receptor (EAAR) antagonist. DOS blocked the depolarizing effects of kainate (KA), but not those of N-methyl-D-aspartate (NMDA) or quisqualate (QQ). Thus DOS was a selective KA antagonist, and KA receptors appear to be the dominant EAAR subtype that mediates synaptic inputs into the inner retina of the mudpuppy. PMID- 2549211 TI - Patterns of intracellular potentials in salamander mitral/tufted cells in response to odor stimulation. AB - 1. Changes in membrane potential and temporal patterns of spikes were analyzed in 30 output cells in the salamander olfactory bulb in response to stimulation with 1-s pulses of the odorants isoamyl acetate, cineole, and camphor. The odor responses were more complex than responses to electrical stimulation of the olfactory nerve or olfactory tracts, with which they were compared. Most began with hyperpolarization and contained prolonged hyperpolarizing and depolarizing potentials that appeared to be compound postsynaptic potentials. These potentials were related to periods of spike inhibition and excitation. The temporal patterns of the responses resembled S-type (for suppression) and E-type (for excitation) patterns described previously in extracellular-unit studies. 2. In single cells, graded but nonmonotonic changes in the responses were observed with increases in the odor concentration from 10(-3) to 10(-1) vapor-phase saturation. Abrupt changes from one category of temporal response pattern to another were generally not observed in response to different concentrations of a single odorant but were frequently observed when the stimulus was changed from one odorant to another. 3. In S-type patterns, the first event was always membrane hyperpolarization and spike inhibition, regardless of the odor concentration. At all concentrations, simple S-type responses were observed in which a single period of hyperpolarization and inhibition lasted several seconds. At moderate to high concentrations, complex S-type responses were observed in which a period of excitation followed an initial period of hyperpolarization and inhibition. In these responses, spikes were often elicited near the termination of the odor pulse, occasionally as early as 300-400 ms after pulse onset. A prolonged period of inhibition followed the period of excitation. 4. In E-type patterns, the first event depended on the odor concentration. At all concentrations, complex responses were observed in which a period of excitation occurred with short latency, followed by a period of inhibition. At low to moderate concentrations, a brief initial period of hyperpolarization preceded the excitation. This initial period of hyperpolarization was always shorter than those in complex S-type responses to equivalent concentrations. However, the range of spike latencies overlapped that of S-type responses to high concentrations. With increasing odor concentration, spike latencies in the E-type responses decreased relative to the onset and peak of the initial hyperpolarization. At high concentrations. spikes were frequently elicited preceding a single period of hyperpolarization and inhibition.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2549212 TI - Serotonergic modulation of two potassium currents in the pleural sensory neurons of Aplysia. AB - 1. The properties of membrane currents that were modulated by serotonin (5-HT) were investigated with two-electrode voltage-clamp techniques in sensory neuron somata isolated from the pleural ganglion of Aplysia californica. The modulatory effects of 5-HT were revealed by computer subtraction of current responses elicited in the presence of 5-HT from current responses elicited prior to the application of 5-HT. The complexities of the resulting 5-HT difference currents (I5-HT) suggested that 5-HT modulated more than one component of membrane current. 2. The 5-HT difference currents appeared to have at least two distinct components. One component was clearly evident at membrane potentials more negative than -10 mV was relatively voltage independent and did not inactivate. A second component was activated at membrane potentials more positive than -10 mV, had complex kinetics, and was highly voltage dependent. In an attempt to identify the membrane currents that were modulated by 5-HT, we compared the pharmacologic sensitivity of I5-HT to that of previously described K+ currents. 3. The two components of I5-HT had different sensitivities to agents that block K+ currents. The relatively voltage-independent component of I5-HT was not blocked by 2 mM 4 aminopyridine (4-AP) and was relatively insensitive to tetraethylammonium (TEA) (estimated Kd of 92 mM). In contrast, the voltage-dependent component of I5-HT was blocked by 4-AP (2 mM) and moderate concentrations of TEA (estimated Kd of 5 mM). 4. The K+ current blockers that were used to examine I5-HT were also used to examine voltage-activated membrane currents. Externally applied TEA blocked the delayed or voltage-dependent K+ current (IK.V) with an estimated dissociation constant (Kd) of 8 mM and a membrane current similar to the Ca2+-activated K+ current (IK.Ca) with an estimated Kd of 0.4 mM. In addition, externally applied 4 AP (2 mM) blocked IK.V. Thus TEA and 4-AP were equipotent in blocking both IK.V and the voltage-dependent component of I5-HT. 5. The suggestion that I5-HT contained multiple components was supported further by examining the modulatory effects of adenosine 3',5'-cyclic monophosphate (cAMP) that mediates some actions of 5-HT on membrane currents in these cells. cAMP difference currents (IcAMP) were similar to the relatively voltage-independent component of I5-HT. The subsequent addition of 5-HT to solutions already containing cAMP resulted in 5-HT difference currents similar to the voltage-dependent component of I5-HT.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2549213 TI - Two components of rapid sensory adaptation in a cockroach mechanoreceptor neuron. AB - 1. The femoral tactile spine of the cockroach is innervated by a single sensory neuron with its cell body in the spine. A step deflection of the spine produces a burst of action potentials which decays to zero in approximately 1 s. This rapid adaptation occurs during encoding of action potentials and can be studied by extracellular electrical stimulation of the sensory neuron. 2. The threshold current of the neuron is labile, increasing with depolarization and decreasing with mild hyperpolarization. During rapid adaptation, the threshold current increases to exceed any steady stimulating current. 3. The dynamic behavior of the threshold current in the tactile spine neuron was observed following step changes in membrane current. The threshold current followed a trajectory which could always be well fitted by a sum of two exponential decays with time constants of approximately 100 and 1,000 ms. 4. The amplitude of the slow component of threshold change was proportional to the size of the step change in membrane current but saturated at strong hyperpolarizations. Its time constant decreased monotonically with depolarization. In contrast, the fast component had more complex behavior, changing biphasically with step amplitude and reversing with initial hyperpolarizations. Its time constant was maximal at the resting membrane potential. 5. The amplitude of the fast component was strongly reduced by treatment with N-chlorosuccinimide (NCS), an agent which reduces sodium channel inactivation. It was not affected by some agents known to block potassium channels. 6. The slow component was not affected by NCS, tetraethyl-ammonium chloride, apamin, or charybdotoxin. 7. Approximate contributions of the two components to the steady-state threshold were reconstructed from step responses. These indicate that the slow component dominates when the receptor is hyperpolarized. However, during depolarizations from rest, as occur with normal stimulation, the two components contribute approximately equally to threshold changes. PMID- 2549214 TI - K+-channel blockade in trigeminal root ganglion neurons: effects on membrane outward currents. AB - 1. The effects of the applications of three K+-channel blockers on the membrane outward currents of neurons were studied with single-electrode voltage-clamp techniques in in vitro slice preparations of the trigeminal root ganglion (TRG) of guinea pigs. The investigations are the first reported attempts to apply these techniques to TRG neurons. 2. During perfusion with tetrodotoxin (TTX; 1 microM), transient outward currents were elicited at the termination of hyperpolarizing voltage commands from holding potentials near -40 mV. The amplitudes of these currents were reduced in conditions of high extracellular [K+]. The activation of such currents was rapid (less than 5 ms), and inactivation was complete at potentials within the activation range. Perfusion with low-[Ca2+], Co2+ containing solutions only slightly and inconsistently reduced the transient outward currents. 3. During combined application of TTX (1 microM) and tetraethylammonium (TEA) (10 mM), fast-activating sustained currents (greater than 1 s) were evoked by depolarizing commands from holding potentials near -70 mV. These currents were blocked completely by the additional inclusion of 5 mM 4 aminopyridine (4-AP) in the perfusing solution. 4. Applications of TEA (0.1-10 mM) produced dose-dependent reductions in the amplitudes of the transient outward currents. Applications of Cs+ also greatly reduced the currents. However, administrations of 4-AP (50 microM-5 mM) diminished these currents only slightly, and high doses of muscarinic agonists had no effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549215 TI - Consequences of 4-aminopyridine applications to trigeminal root ganglion neurons. AB - 1. The effects of 4-aminopyridine (4-AP) on the electrical properties of 30 trigeminal root ganglion (TRG) neurons were determined from the membrane voltage responses to step and sinusoidal current injections using intracellular microelectrode techniques in in vitro slice preparations (guinea pigs). 2. Comparisons of results from 4-AP applications (0.05-5 mM) with those from tetraethylammonium (TEA) applications (0.1-10 mM) revealed very different actions of these agents. Both agents produced an increase in input resistance and a decrease in threshold for spike generation. Applications of 4-AP increased subthreshold oscillations of the membrane potential and enhanced the repetitive spike firing evoked by intracellular injections of current pulses. However, TEA applications blocked the potential oscillations and did not exaggerate repetitive spike discharges. Spontaneous spike activity or bursts were observed in four neurons that received 4-AP applications. 3. Membrane properties were determined in 20 of the 30 neurons by fitting impedance data in the frequency domain with a four-parameter membrane model by the use of computer-intensive techniques. In the majority of neurons, the time-invariant and time-dependent membrane conductances decreased during 4-AP application. The time constant for the time-dependent conductance also decreased, suggesting that the closing of K+-channels was facilitated in the membrane. 4. Applications of 4-AP in a dose range of 50 microM 5 mM produced rapid (approximately tens of seconds) responses of the neurons, resulting in a dose-dependent increase of the impedance magnitude functions and in a leftward shift of the resonant "humps" to lower frequencies. This shift indicates that the TRG neuronal membrane is capable of producing large voltage responses to current inputs at low frequencies. Recovery from the effects of 4-AP was slow (usually greater than 30 min). 5. Applications of 4-AP at high doses (greater than or equal to 1 mM) and at various imposed membrane potentials in four neurons resulted in poorly reversible unspecific changes in certain membrane parameters (increased input capacitance and conductance) and an insensitivity of the input conductance to the imposed membrane potential. These effects could be interpreted as membrane breakdown. 6. The tendencies of TRG neurons to fire repetitively and in bursts of spikes during 4-AP application result from the increased oscillatory behavior of their membrane potentials and changes in membrane resonance induced by presumed blockade of K+ channels.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2549216 TI - Neuropeptide Y receptor binding sites in rat brain: differential autoradiographic localizations with 125I-peptide YY and 125I-neuropeptide Y imply receptor heterogeneity. AB - Neuropeptide Y (NPY) receptor binding sites have been localized in the rat brain by in vitro autoradiography using picomolar concentrations of both 125I-NPY and 125I-peptide YY (PYY) and new evidence provided for differentially localized receptor subtypes. Equilibrium binding studies using membranes indicate that rat brain contains a small population of high-affinity binding sites and a large population of moderate-affinity binding sites. 125I-PYY (10 pM) is selective for high-affinity binding sites (KD = 23 pM), whereas 10 pM 125I-NPY labels both high and moderate-affinity sites (KD = 54 pM and 920 pM). The peptide specificity and affinity of these ligands in autoradiographic experiments match those seen in homogenates. Binding sites for 125I-PYY are most concentrated in the lateral septum, stratum oriens, and radiatum of the hippocampus, amygdala, piriform cortex, entorhinal cortex, several thalamic nuclei, including the reuniens and lateral posterior nuclei, and substantia nigra, pars compacta, and pars lateralis. In the brain stem, 125I-PYY sites are densest in a variety of nuclei on the floor of the fourth ventricle, including the pontine central grey, the supragenual nucleus, and the area postrema. 125I-NPY binding sites are found in similar areas, but relative levels of NPY binding and PYY binding differ regionally, suggesting differences in sites labeled by the two ligands. These receptor localizations resemble the distribution of endogenous NPY in some areas, but others, such as the hypothalamus, contain NPY immunoreactivity but few binding sites. PMID- 2549217 TI - Extracellular cAMP accumulation and degradation in rat cerebral cortex in dissociated cell culture. AB - Norepinephrine (NE) stimulated the accumulation of cAMP in embryonic rat cerebral cortex in dissociated cell culture. After exposure to NE for 10 min, the intracellular cAMP content of these cultures went from 22 +/- 12 to 202 +/- 75 pmol/mg protein. Using selective culturing techniques, evidence was obtained supporting the hypothesis that NE-stimulated production of cAMP is a property associated with the glial rather than the neuronal component of these cultures. Beta adrenergic agonist stimulation of cortical cultures also resulted in the efflux of cAMP into the medium. At the peak of extracellular accumulation of cAMP (following a 40-min exposure to isoproterenol), 180 pmol cAMP/mg protein had been transported into the extracellular medium. The fate of extracellular cAMP was investigated using thin-layer chromatography. Extracellular cAMP was degraded to AMP and adenosine; this degradation did not seem to be due to the presence of serum or serum components, suggesting the existence of an extracellular phosphodiesterase. In response to NE stimulation of glia, in particular astrocytes, cAMP or its metabolites may accumulate at high enough concentrations in the extracellular space in cerebral cortex to affect neuronal function, possibly via adenosine receptors. PMID- 2549218 TI - Endogenous activation of glycine and NMDA receptors in lamprey spinal cord during fictive locomotion. AB - Strychnine is shown to abolish left-right alternation in fictive locomotion induced by sensory stimulation. Robust rhythmic activity, characterized by left right coactivation at each segmental level, is seen in the presence of strychnine at all doses used (0.5-20 microM). The proportion of the cycle occupied by the ventral root bursts and the rostral-caudal coordination is similar to that seen in the absence of strychnine. Furthermore, the rhythm is abolished by cis-2,3 piperidine dicarboxylic acid (PDA), 2-amino-5-phosphonovalerate (APV), or the removal of Mg2+ from the perfusate, as in the absence of strychnine. Voltage clamp was applied to ventral horn neurons during stimulation in the presence of strychnine, with holding potentials negative to the plateau potential associated with a ventral root burst but positive to the potential in the interburst. Inward current was seen during the ventral root burst, but no outward current was seen at burst termination or during the interburst. The results indicate that in fictive locomotion induced by endogenous release of NMDA receptor agonists, left right alternation is dependent on glycinergic transmission. Furthermore, evidence is provided that in the absence of glycinergic transmission, burst termination may depend on NMDA receptor-linked voltage-sensitive processes. PMID- 2549219 TI - Differential ascending projections to aural regions in the 60 kHz contour of the mustache bat's inferior colliculus. AB - The inferior colliculus of the mustache bat is similar in many respects to the inferior colliculus of more commonly studied mammals. However, the isofrequency contour devoted to processing 60 kHz, the dorsoposterior division (DPD) is greatly expanded, encompassing an area approximately equal to one-third of the central nucleus. Of particular significance is that monaural and binaural neurons are segregated in the DPD into 4 spatially distinct aural regions. In this study we exploit the great enlargement of the 60 kHz region in the central nucleus of the inferior colliculus (ICc) of the mustache bat to determine the source of ascending projections to the 4 different aural regions of the DPD. Small iontophoretic deposits of HRP were made within each of the physiologically defined aural regions, and the locations and numbers of retrogradely labeled cells in the auditory brainstem nuclei were determined. Two major features of collicular organization were found. The first is that each aural region receives a unique set of projections from a subset of lower auditory nuclei and thus is distinguished both by its neural response properties and by the pattern of ascending projections it receives. The dorsomedial EE region receives inputs primarily from the ipsilateral intermediate nucleus of the lateral lemniscus (INLL) and ventral nucleus of the lateral lemniscus (VNLL), and the contralateral ICc. In contrast, the ventrolateral EE region receives projections from the ipsilateral medial superior olivary nucleus (MSO), VNLL, and INLL. The inputs to the EI region originate primarily from the dorsal nucleus of the lateral lemniscus (DNLL) and lateral superior olivary nucleus (LSO) bilaterally and from the ipsilateral INLL. The afferents to the EO region include the contralateral cochlear nucleus, the ipsilateral VNLL and INLL and MSO. The second major organizational feature is that the binaural nuclei of the brain-stem project upon the DPD in a more restricted manner than do some of the lower monaural nuclei, such as the VNLL and INLL, which project in a more widespread manner. The unique set of projections terminating in each aural region of the DPD suggests that the neurons should have substantially different properties, even when neurons in different regions are of the same general aural type. Moreover, the elucidation of the micro-organization of the DPD provides insights into the different ways that binaural properties of DPD neurons are created by the convergence of inputs from particular subsets of lower auditory nuclei.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2549220 TI - GABA regulation of circadian responses to light. I. Involvement of GABAA benzodiazepine and GABAB receptors. AB - Light-induced phase shifts of the circadian locomotor rhythm of hamsters can be blocked by agents that alter GABA neurotransmission. The GABA antagonist bicuculline blocks phase delays induced by light and the benzodiazepine diazepam, which can potentiate GABA activity, blocks light-induced phase advances. In the experiments reported here, we found that the bicuculline blockade of phase delays was reduced by agents that mimic or potentiate GABA activity. Conversely, the diazepam blockade of phase advances was reduced by both competitive and noncompetitive antagonists of GABA. This indicates that the GABA-benzodiazepine receptor-ionophore complex is the most likely site of action for the effects of these drugs on circadian rhythms. However, competitive GABA agonists did not mimic the blocking effects of benzodiazepines, nor did the antagonist picrotoxin mimic the blocking effect of bicuculline. Therefore, the classic action of GABA, increased chloride conductance, may not be the effector mechanism in this case. We also found that the GABAB agonist baclofen blocked both phase advances and delays and that the blockade of advances was reversed by the antagonist delta aminovaleric acid. Taken together, these results indicate that GABA is involved in the regulation of circadian responses to light and that the regulation is mediated by both GABAA and GABAB receptors. PMID- 2549221 TI - Neurotransmission regulates stability of acetylcholine receptors at the neuromuscular junction. AB - The majority of acetylcholine receptors (AChRs) at normally innervated neuromuscular junctions are stable, with a half-life averaging about 12 d in most rodent muscles. Following denervation, the AChRs turn over much more rapidly after a lag period. The mechanism by which motor nerves normally maintain stabilization of junctional AChRs is not yet known. In order to determine whether synaptic transmission plays a role in this process, we have compared the effects of pre-and postsynaptic chloinergic blockade with those of surgical denervation. 125l-alpha-bungarotoxin was used to label junctional AChRs and follow their loss over time. Presynaptic blockade of quantal ACh transmission was produced in the soleus (SOL) and flexor digitorum brevis muscles of mice by repeated injections of type A botulinum toxin. Postsynaptic blockade of quantal and nonquantal ACh transmission was produced by continuous infusion of alpha-bungarotoxin in the SOL. Our findings show that treatment with botulinum toxin resulted in an accelerated loss of junctional AChRs that was similar to the effects of surgical denervation, though briefly delayed in its onset. Treatment with alpha bungarotoxin produced an effect that was quantitatively equivalent to the accelerated loss of junctional AChRs following surgical denervation, with an identical time course. These results support the concept that cholinergic transmission is a mediator of the neural control of stability of junctional AChRs. The possibility that receptor stabilization may represent a mechanism of long-term postsynaptic "memory" dependent on neural transmission is discussed. PMID- 2549222 TI - Prognostic implications of the bromodeoxyuridine labeling index of human gliomas. AB - This study includes 182 patients with intracranial gliomas who received bromodeoxyuridine (BUdR), 200 mg/sq m intravenously, at the time of craniotomy but before tumor biopsy. The tumor specimens were stained for BUdR using the immunoperoxidase method; the BUdR labeling index (LI), or S-phase fraction, was calculated as the percentage of BUdR-positive cells. The median BUdR LI's for 127 primary moderately anaplastic astrocytomas, highly anaplastic astrocytomas, and glioblastomas (less than 1%, 2.7%, and 7.3%, respectively; range 0% to 38.1%) were not significantly different from those of 55 similar recurrent tumors (less than 1%, 4.3%, and 7.4%, respectively; range 0% to 30.5%). The mean LI was significantly higher in tumors from patients over 50 years of age than in tumors from younger patients (p less than 0.001). The age-related difference in LI's was found in both groups of patients with astrocytomas but not in those with glioblastomas. Kaplan-Meier survival curves showed a significantly greater probability of survival among patients whose tumors had LI's of less than 1% than among those with LI's greater than 5%; survival probability of patients with tumor LI's of 1% to 5% was intermediate between the two extremes. Thus, the BUdR LI appears to reflect the proliferative potential more accurately than the histopathological diagnosis and should therefore be considered an important factor in determining the prognosis of individual patients with intracranial gliomas and in selecting their treatment. PMID- 2549223 TI - Use of thallium-201 SPECT to quantitate malignancy grade of gliomas. AB - A quantitative preoperative technique using thallium-201 single-photon emission computerized tomography is described which predicts whether specific gliomas are of high- or low-grade malignancy. An index, based on the ratio of thallium uptake in the tumor versus the homologous contralateral brain, was calculated and compared with tumor histology. The index in 14 patients with low-grade malignant gliomas was 1.27 +/- 0.40 in contrast to an index of 2.40 +/- 0.61 in 11 patients with high-grade malignant gliomas (p less than 0.0005). Whether gliomas were of low- or high-grade malignancy could be predicted with 89% accuracy using a threshold of 1.5. Low-grade gliomas with an index higher than 1.5 acted biologically more like high-grade tumors, and no tumor histologically classified as being of high-grade malignancy had an index lower than 1.7. This technique could help to reduce unrecognized sampling errors during needle biopsies of brain tumors, particularly of high-grade lesions classified in error as low-grade tumors due to inadequate biopsy material. PMID- 2549224 TI - Changes in xanthine oxidase in ischemic rat brain. AB - Xanthine oxidase activity in the rat brain was measured by means of high performance liquid chromatography with electrochemical detection of uric acid. Cerebral ischemia was produced by a four-vessel occlusion method. In the control rat, the enzyme activity was 0.87 +/- 0.13 nmol/gm wet weight/min at 25 degrees C (mean +/- standard deviation), of which 92.4% was associated with the nicotinamide adenine dinucleotide (NAD)-dependent dehydrogenase form and only 7.6% with the oxygen-dependent superoxide-producing oxidase form. However, the ratio of the latter form increased to 43.7% after 30 minutes of global ischemia, despite the total xanthine oxidase activity remaining the same. Thus, it was revealed that uric acid can be synthesized in the rat brain and that cerebral ischemia induced the conversion of xanthine oxidase from an NAD-dependent dehydrogenase to an oxygen-dependent superoxide-producing oxidase. Although the xanthine oxidase pathway has been proposed as a source of oxygen-derived free radicals in various ischemic organs other than brain, the results of the present study suggest the involvement of the oxygen free radicals generated from this pathway in the pathogenesis of the ischemic injury of the rat brain. PMID- 2549226 TI - [Intranasal infection of ICR mice with herpes simplex virus type 1]. AB - Intranasal inoculation of mice with herpes simplex virus (HSV) provides a model of human herpetic infection through a natural route of inoculation. Five-week-old male ICR mice were infected intranasally with various strains of herpes simplex virus type 1 (HSV-1), and the fundamental aspects of the pathogenicity of this virus were studied. Six virus strains examined showed variance in their virulence determined by lethal dose 50 (LD50) for mice. Four of the strains were revealed to be virulent, and two were shown to be attenuated. The relative degree of virulence among these strains corresponded well to that shown by intraperitoneal inoculation. When mice were inoculated with a virulent virus strain (F), virus multiplication was shown clearly in several organs tested, such as the lung, brain, olfactory bulb, trigeminal ganglion, spinal cord and adrenal gland. Viremia was also demonstrated. On the other hand, in mice inoculated with an attenuated virus strain (-GCr), virus was recovered only from the lung and adrenal gland and in much less amount than in the respective organs of mice infected with the virulent strain. No viremia was demonstrated. These data strongly suggest that the lethal effect of HSV-1 on mice is dependent upon whether or not significant virus multiplication occurs in the central nervous system, which is the critical target organ of HSV. Preinoculation of mice with an attenuated strain via the intranasal route suppressed the lethal effect of subsequent infection with any of the virulent strains by the same route of inoculation, although this protection phenomenon was not so pronounced when the virulent UNO-1A strain was used.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549225 TI - Kinetic sensitivity of a receptor-binding radiopharmaceutical: technetium-99m galactosyl-neoglycoalbumin. AB - Kinetic sensitivity is the ability of a physiochemical parameter to alter the time-activity curve of a radiotracer. The kinetic sensitivity of liver and blood time-activity data resulting from a single bolus injection of [99mTc]galactosyl neoglycoalbumin [( Tc]NGA) into healthy pigs was examined. Three parameters, hepatic plasma flow scaled as flow per plasma volume, ligand-receptor affinity, and total receptor concentration, were tested using [Tc]NGA injections of various molar doses and affinities. Simultaneous measurements of plasma volume (iodine 125 human serum albumin dilution), and hepatic plasma flow (indocyanine green extraction) were performed during 12 [Tc]NGA studies. Paired data sets demonstrated differences (P(chi v2) less than 0.01) in liver and blood time activity curves in response to changes in each of the tested parameters. We conclude that the [Tc]NGA radiopharmacokinetic system is therefore sensitive to hepatic plasma flow, ligand-receptor affinity, and receptor concentration. In vivo demonstration of kinetic sensitivity permits delineation of the physiologic parameters that determine the biodistribution of a radiopharmaceutical. This delineation is a prerequisite to a valid analytic assessment of receptor biochemistry via kinetic modeling. PMID- 2549227 TI - [A study on collagenase production of nasal polyp-derived fibroblasts stimulated by nasal secretions of chronic sinusitis]. AB - The collagenase produced by mesenchymal cells has been thought to have a great importance in the pathophysiology of connective tissue metabolism and prolongation of chronic inflammation. The factors, such as IL-1 and PMN factor, released by inflammatory cells have been known to induce mesenchymal cells to produce collagenase. In the present study, the collagenase activity of the nasal secretions were estimated using FITC-labelled collagens as substrates. The factor, enhancing the fibroblasts to produce collagenase, was also isolated from nasal secretions and partially characterized. The fibroblasts used in the present study were cultured with explant of the sections of nasal polyp obtained from a patient with chronic sinusitis. The collagenase activity in nasal secretions from patients with chronic sinusitis was high, whereas that of allergic nasal secretions was extremely low. Furthermore, the collagenase productions of nasal polyp-derived fibroblasts were enhanced by the extracts of nasal secretions from patients with chronic sinusitis. Crude extracts of nasal secretions were fractionated by ammonium sulfate precipitation. The active materials precipitated by 50% to 80% ammonium sulfate were further purified by Sephadex G-75 gel chromatography. The molecular weight determination of the active fraction checked by HPLC utilizing for TSK 2,000 SW gel column indicates 20,000 daltons for the active materials. However, the collagenase production of human microvascular endothelial cells derived from nasal mucosa was not enhanced by this factor. Although either the origin or the nature was not confirmed, the factor was considered to relate to the prolongation of chronic inflammation in the nasal and paranasal sinus pathology. Analysis of these factors will expected to establish methods for new therapeutics in chronic inflammation. PMID- 2549229 TI - A strategy for demonstrating the clonal origin of small numbers of T lymphocytes in histopathological specimens. AB - A strategy is described for detecting large T-cell clones among small numbers of lymphoid cells in fresh or formalin-fixed tissue samples using the polymerase chain reaction (PCR) to amplify and identify rearrangements of the V and J genes of the T-cell gamma receptor. Following hybridization of primers with gene specific sequences at judiciously selected locations on each of the eight potentially active V and five potentially active J genes, the PCR can theoretically amplify DNA segments which span the join between rearranged V and J genes and are of approximately 384 different sizes, each segment size reflecting different gamma gene clonal rearrangements. Large monoclonal populations of T lymphocytes, indicated by excessive amounts of particular PCR segment sizes, can be further characterized by direct nucleotide sequencing of the hypervariable N regions of these segments, and the presence of such clones can be confirmed directly in tissue sections by in situ hybridization with N region-specific oligonucleotide probes. PMID- 2549228 TI - Immunoreactivity for laminin and type IV collagen in malignant and benign fibrous histiocytoma. AB - Sixteen malignant fibrous histiocytomas (MFH) and ten benign fibrous histiocytomas of the skin were studied immunohistochemically for the distribution of two basement membrane (BM) proteins, laminin and type IV collagen, in order to evaluate their cellular nature. Linear staining for both proteins was present in the vascular BMs. Intracytoplasmic laminin was observed in the neoplastic fibroblast-like and pleomorphic giant cells in 11 MFHs. Two MFHs also showed similar staining for type IV collagen. In the giant cell subtype of MFH, the reactive giant cells were totally negative whereas the neoplastic cells were strongly positive for laminin. Extracellular fibres staining positively for both BM proteins were seen in two MFHs. Except for the capillary network, the benign fibrous histiocytomas were negative for laminin and type IV collagen. On the basis of the present results, we favour the concept that MFHs are primitive mesenchymal tumours, some of which may show histogenetic relationships with tumours of BM forming mesenchymal cells. PMID- 2549230 TI - Structural modifications of lipid vesicles by 8-methoxypsoralen. AB - On prolonged UV-A illumination the ESR spectrum of 16-doxylstearic acid in dimyristoyl phosphatidylcholine vesicles loaded with 8-methoxypsoralen changed dramatically as a second broad component gradually appeared. The composite nature of the spectrum was proven by digital subtraction and the two components were simulated using the modified Bloch equations. From the spectral anisotropy of the motionally averaged three-line component, the molecular order was found to decrease during UV-A illumination, and isotropic rotation of the spin probe was observed well below the phase transition temperature. In parallel an exchange averaged component dominated the central part of the spectrum, indicating the segregation of spin probes into separate domains. The build-up of the composite lineshape was followed by fitting weighted sums of the above two components to spectra recorded after various time intervals. It is concluded that PUVA therapy, which is routinely used against psoriasis, might also induce overlooked damage to the saturated lipid species of biological membranes. PMID- 2549231 TI - Epidemiology and laboratory diagnosis of infection with viral and bacterial pathogens in infants hospitalized for suspected sepsis. AB - A prospective study was conducted to determine the frequency and distribution of bacterial and viral pathogens in infants hospitalized with suspected sepsis and to evaluate the potential of virus detection for improving patient management. A causative organism was detected in 157 (67%) of 233 previously healthy infants less than 3 months of age, who had been hospitalized for suspected sepsis: 19 (8%) had bacterial infections, 135 (58%) had viral infections, and 3 (1%) had mixed viral-bacterial infections. Viral infections occurred in a seasonal pattern: enteroviruses were responsible for most of the hospitalizations during summer and fall (65/110; 63%) and respiratory syncytial and influenza A viruses were responsible for most of the infections during winter (44/81; 55%). In contrast, bacterial infections were not seasonally distributed. Virus was detected in 33% of the 138 infected infants within 24 hours, and in 64% within 3 days. We conclude that viral infections are prevalent among infants hospitalized for suspected sepsis, and most can be detected early enough to influence patient management. PMID- 2549232 TI - Surgical treatment of thoracic deformity in Poland's syndrome. AB - In 1841, Poland described congenital deficiency of the pectoralis major and minor muscles associated with syndactyly. This syndrome is a spectrum, often involving chest wall and breast deformity as well. Identification of the various musculoskeletal components involved permits optimal thoracic reconstruction in the small proportion of patients who will require it. From 1955 to 1988, 75 patients (40 males and 35 females) with Poland's syndrome were treated or evaluated. Patients with isolated deficiencies of the pectoral muscles, breast, or hand deformity were excluded. The complex was right-sided in 44 patients, left sided in 30, and bilateral in one. The pectoralis minor and the costal portion of the pectoralis major muscle were absent in all patients. Hand anomalies were present in 50 patients. Athelia and/or amastia were noted in 37 patients. In ten patients, the rib cage deformity required reconstruction, and in three cases, rib or cartilage grafts were needed for complete repair. Often unappreciated in these cases is the significant rotation of the sternum toward the involved side and contralateral carinate deformity. Correction is achieved by bilateral subperichondrial costal cartilage resection and sternal osteotomy (seven of ten patients), thus allowing anterior displacement and orthorotation of the sternum. Chest wall reconstruction must be tailored to the requirements of each patient. No intraoperative or postoperative complications occurred in these ten patients. In males without rib cage deformity, generally no treatment is required to replace the absent pectoral muscles, although in two cases rotation of the latissimus dorsi muscle was performed. In all females, reconstruction of the ipsilateral breast is required at full development.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549233 TI - Analysis of transforming growth factor beta receptor binding in embryonic, fetal, and adult rabbit fibroblasts. AB - Adult wound repair traits including inflammation, fibroplasia, and collagen deposition are not seen at fetal wound sites. This observation raised questions about regulatory mechanisms extant in fetal healing. Transforming growth factor beta (TGF-beta) is an important regulatory polypeptide known to orchestrate fibroplasia and collagen synthesis during adult wound repair. Previous studies have suggested that the wounded rabbit fetus is capable of responding with these adult characteristics if provided with exogenous TGF-beta. In order to test whether the observed in vivo effects of TGF-beta in the rabbit fetus might be due to a direct effect on the fibroblast, TGF-beta receptor binding characteristics of early passage cultured embryonic (14 days' gestation), fetal (24 days' gestation), and adult rabbit fibroblasts were studied by flow cytometry. Experiments were carried out using fluorescein-conjugated TGF-beta (F-TGF-beta) with analysis on an EPICS V flow cytometer. F-TGF-beta was incubated with each of the three fibroblast types at 37 degrees C after which time the cells were washed twice and analyzed with a minimum of 10(5) cells for each data point. F-TGF-beta bound rapidly and reversibly to the embryonic, fetal, and adult fibroblasts with saturation being achieved at 1 nmol/L for fetal and adult cells, and 8 nmol/L in the embryonic fibroblasts. Saturating concentrations of F-TGF-beta yielded mean channel numbers (a function of relative amounts of F-TGF-beta-bound) of 172, 114, and 97 for embryonic, fetal, and adult cells, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549234 TI - Unusual ultrastructural features in the superficial pineal gland of the yellow bellied country rat (Rattus losea Swinhoe). AB - The ultrastructure of the superficial pineal gland of the yellow-bellied country rat (Rattus losea Swinhoe), living under natural climatic conditions at the margin of the tropical region, was studied with special reference to the seasonal changes. In the rainy season with average monthly rainfall of 400 mm (light phase of 13.5 h, and temperature of 28 degrees C) the gland was characterized by the presence of frequent large vacuoles containing flocculent material (LVFs), large condensed inclusions (LCIs) in the pinealocytes, and extensive accumulations of presumptive secretion in the extracellular compartment, suggesting an enhanced secretory activity. In the dry season with average monthly rainfall of 13 mm (light phase of 11 h, and temperature of 19 degrees C), however, the pinealocytes were characterized by the presence of numerous clear spindles, which exhibited acid phosphatase activity; at this time there were few LVFs and LCIs and rare extracellular accumulations, suggesting a declined glandular activity. We conclude that the cellular activity of the pineal gland in R. losea, living under tropical climatic conditions, is synchronized with the annual changes of rainfall, which is apparently more important than the annual changes of photoperiod and temperature. PMID- 2549235 TI - Snuff tumorigenesis: effects of long-term snuff administration after initiation with 4-nitroquinoline-N-oxide and herpes simplex virus type 1. AB - The tumor promoting effects of snuff was studied in Lewis rats initiated with 4 nitroquinoline-N-oxide (4-NQO) and Sprague Dawley rats repeatedly inoculated with herpes simplex virus type 1 (HSV-1). The test substances were administered in a surgically created canal in the lower lips of the rats. There were 15 rats in each test group and 10 rats in the control group. In the groups treated with 4 NQO and 4-NQO + snuff, 8 and 12 tumors (5 and 9 malignant) were found, respectively. In the group subjected to HSV-1 only, 3 tumors were found (2 malignant), in the group subjected to snuff only, 4 tumors were found (3 malignant) and in the group subjected to the combination of HSV-1 and snuff, 13 tumors were found (7 malignant). In the control group only one malignancy was found. The study did not show any promoting effects of snuff in the oral cavity after initiation with 4-NQO. Neither was there any increase in the number of oral tumors in rats treated with HSV-1 and snuff. However, there was a marked increase in the number of malignant tumors outside the oral cavity in the group treated with HSV-1 and snuff, underlining the importance of interactions between these two agents in the development of malignant lesions. PMID- 2549236 TI - Regional odontodysplasia: a bibliography. AB - Regional odontodysplasia is a localized disorder of tissues of dental origin resulting in characteristically bizarre clinical and radiographic appearances. It most commonly affects the maxillary anterior teeth of both the permanent and primary dentitions. The authors present a worldwide bibliography of reported cases and discuss the suggested theories regarding the condition's etiology. Although the majority of cases of regional odontodysplasia must be regarded as idiopathic, various developmental anomalies may produce identical changes in the teeth in one or more quadrants. A number of previously reported cases are also reconsidered with regard to their diagnosis and guidelines are suggested for the treatment of patients affected by regional odontodysplasia. PMID- 2549237 TI - Characterization of muscarinic receptors: type M2 (subtype B) on neuro-2A neuroblastoma cells. AB - The binding of the nonselective muscarinic antagonist, [3H]N-methylscopolamine (NMS) to a mouse neuroblastoma cell line (Neuro-2A) and its coupling to the inhibition of adenylate cyclase were characterized. Specific [3H]NMS binding to membrane preparations was rapid, saturable, and of high affinity. Saturation experiments revealed a single class of binding sites for the radioligand. Competition experiments with the muscarinic drugs pirenzepine, AF DX 116, dicyclomine and atropine revealed that the muscarinic receptors present on these cells are predominantly of a single class, subtype B (M2). In addition, agonist binding demonstrated existence of a GTP-sensitive high affinity binding state of the receptors. Coupling of these muscarinic receptors to the adenylate cyclase system was investigated using the muscarinic agonist carbachol which was able to inhibit the prostaglandin (PGE1)-stimulated activation of adenylate cyclase. The agonist carbachol did not stimulate the formation of IP3 above basal levels, which indicated that the receptors are not coupled to phosphatidylinositol metabolism. In conclusion, we show that possessing predominantly one subtype of muscarinic receptor, the Neuro-2A cells provide a useful model for the investigation of the heterogeneity of muscarinic receptors and the relationship of subtype to the coupling of different effectors. PMID- 2549238 TI - Value of intrauterine device insertion and estrogen administration after hysteroscopic metroplasty. AB - Hysteroscopic metroplasty seems the treatment of choice for septate uterus. Little information is available on the possible usefulness of postoperative intrauterine device (IUD) insertion and estrogen administration in preventing fusion of a freshly cut septum and intrauterine adhesion formation. A hysteroscopic incision in a uterine septum was made in 20 women. Postoperatively an IUD was inserted in ten of them and conjugated estrogens administered for 30 days with medroxyprogesterone acetate on days 26-30 (group I); the other ten were given no other therapeutic measures (group II). On follow-up hysterography five women in group I had a normal uterine cavity, and five had a residual fundal notch greater than or equal to 1 cm. In group II four had a normal uterine cavity, and six had a residual fundal notch greater than or equal to 1 cm. No intrauterine adhesions were detected in any of the patients. IUD insertion and hormonal therapy after hysteroscopic metroplasty do not seem to be needed to prevent septal fusion. PMID- 2549239 TI - Small cell carcinoma of the vagina with neuroendocrine features. A report of three cases. AB - Three patients were treated for primary small cell carcinoma of the vagina. The light microscopic features were similar to those of small cell carcinoma of the lung, cervix and endometrium. Electron microscopy revealed occasional cytoplasmic, neurosecretory-type granules and cytoplasmic processes, features consistent with neuroendocrine cells. Radiotherapy appears to provide local control, but widespread disease at presentation or early metastases suggest the use of adjuvant chemotherapy in the initial management of the neoplasm. PMID- 2549240 TI - Gold(I) compounds and human neutrophil collagenase. PMID- 2549241 TI - Increment of Tal positive cells in peripheral blood from patients with rheumatoid arthritis. AB - Recently it was demonstrated that the Tal antigen is a marker for activated T cells and the population of Tal+ cells includes memory T cells. Our study was undertaken to analyze Tal+ cells in peripheral blood from patients with rheumatoid arthritis (RA) and examine the correlation between the proportion of Tal+ cells and disease activity. Using a dual immunofluorescent method, T cell subsets were analyzed in peripheral blood from 46 patients with RA, 10 patients with osteoarthritis (OA) and 10 healthy subjects. Patients with RA had a significantly higher percentage of T cells bearing the Tal antigen than patients with OA and healthy subjects. Furthermore, the percentage of CD3+Tal+ cells in peripheral blood from patients with RA was significantly correlated with disease activity including erythrocyte sedimentation rate, morning stiffness and the Lansbury index, whereas CD3+ HLA-DR+ cells did not correlate. In serial observations in patients with RA, the percentage of CD3+ Tal+ cells decreased in accordance with the improvement of disease activity. Thus, we suggest that the percentage of Tal+ cells may be a marker of the disease activity in patients with RA. PMID- 2549242 TI - Immunoglobulin G coating on crystals and ceramics enhances polymorphonuclear cell superoxide production: correlation with immunoglobulin G adsorbed. AB - Crystals and a group of ceramics used in joint replacement surgery were incubated with polymorphonuclear cells (PMN) with and without adsorbed immunoglobulin G (IgG). All 11 particles tested had the capacity to induce the production of superoxide anion (O2-) to at least some extent. Synthetic calcium pyrophosphate dihydrate crystals (CPPD) and several ceramics adsorbed small amounts of IgG and the increment of superoxide production was also small. Both the augmentation of O2- and the adsorption of IgG to hydroxyapatite type I synthetic crystals and the remaining ceramics were intermediate. Synthetic monosodium urate crystals adsorbed the most IgG and the O2- increments were large. The increase in superoxide production correlated with the amount of IgG bound onto crystals or ceramics. The greater protein binding and phlogistic potential of some ceramics needs to be taken into consideration in their clinical use. PMID- 2549243 TI - Randomised blind controlled trial of a high fibre, low fat and low sodium dietary regimen in mild essential hypertension. AB - Thirty-four patients with essential hypertension were allocated, in a controlled trial, to a treatment diet of high fibre, low fat and low sodium composition, or to a control diet by the hospital dietitian. Clinical observations were made by a separate 'blinded' nursing sister. After three months treatment, the modified diet-treated group showed a significant reduction in mean systolic (169.4 +/- 23.4 to 150.6 +/- 16.1 mmHg) and diastolic blood pressure (101.5 +/- 7.3 to 89.4 +/- 6.8 mmHg), accompanied by significant reductions in urinary sodium excretion (140.4 +/- 34.6 to 93.7 +/- 44 mmol/day) and weight (73.1 +/- 10 to 71.2 +/- 8.4 kg). The changes in control were; systolic 171.2 +/- 14.1 to 162.1 +/- 19.5 mmHg and diastolic pressure 97.2 +/- 10.8 to 91.7 +/- 9.7 mmHg. The mean differences in reductions between treated and control were 8.8 mmHg Systolic (95% confidence intervals: -2.6 to 21.2 mmHg) and 7.0 mmHg diastolic blood pressure (95% confidence intervals: 0.4 to 14.4 mmHg). The number of patients with normal blood pressure in the diet treated group at three months was double that in the control (eleven versus five). No relationships were shown between blood pressure changes and those of weight or urinary sodium excretion during the trial. The findings in this study are broadly in agreement with similar ones in essential hypertension and suggest that this form of dietary regimen has a clinically worthwhile hypotensive effect and this should be readily achievable in routine clinical practice. PMID- 2549244 TI - LH-RH antagonists: design and synthesis of a novel series of peptidomimetics. PMID- 2549245 TI - Evaluation and synthesis of aminohydroxyisoxazoles and pyrazoles as potential glycine agonists. AB - Except for structurally similar small amino acids, such as alanine, beta-alanine, and serine, compounds acting as glycine-receptor agonists are an unknown class of pharmacological agents. To investigate the potential of small, substituted heterocycles to act as glycine agonists, we have evaluated the similarities between glycine and a series of hydroxy- and amino-substituted pyrazoles and isoxazoles through complementary molecular modeling techniques. Using a "scorecard approach" to determine the overall similarity of projected agonist structures to glycine, we prioritized synthesis and subsequently prepared several novel derivatives. The biological activity of these compounds was compared to that of glycine by using a [3H]strychnine-mediated glycine receptor binding assay. Despite the close similarity in the calculated parameters when compared to glycine, no significant receptor-binding activity was observed for the targeted analogues. These results illustrate the structurally exacting nature of the glycine receptor. PMID- 2549246 TI - 4-(Phosphonoalkyl)- and 4-(phosphonoalkenyl)-2-piperidinecarboxylic acids: synthesis, activity at N-methyl-D-aspartic acid receptors, and anticonvulsant activity. AB - A series of 4-(phosphonoalkyl)- and 4-(phosphonoalkenyl)-2-piperidinecarboxylic acids were synthesized, and their biological activity was assessed as competitive ligands for the NMDA receptor, both in vitro by using a receptor binding assay ([3H]CGS 19755 binding) and in vivo by using an NMDA seizure model in mice. The analogues were also evaluated in [3H]AMPA and [3H]kainate binding to assess their affinity for non-NMDA excitatory amino acid receptor subtypes. A number of these analogues show potent and selective NMDA antagonistic activity both in vitro and in vivo. Most notable are 4-(phosphonomethyl)-2-piperidinecarboxylic acid (1a) (CGS 19755) and the phosphonopropenyl analogue 1i, both of which show anticonvulsant activity in the 1-2 mg/kg ip range. With the aid of computer assisted modeling, a putative bioactive conformation for AP-5 is hypothesized from the SAR data presented and a preliminary model for the antagonist-preferring state of the NMDA receptor is presented. PMID- 2549247 TI - Benzofuro[2,3-c]pyridin-6-ols: synthesis, affinity for opioid-receptor subtypes, and antinociceptive activity. AB - A general synthetic approach to a novel series of cis-1,2,3,4,4a,9a hexahydrobenzofuro[2,3-c]pyridin-6-ols is described together with their receptor binding profile on opioid-receptor subtypes (mu, kappa, delta). In addition, their in vivo antinociceptive activity was assessed. A number of the analogues synthesized showed potent affinity for opioid receptors and have potent antinociceptive activity in a mouse phenylquinone abdominal stretching model. In addition, the SAR for nitrogen substitution in the above series is explored with respect to the overall opioid receptor subtype binding profile. In general it was found that substituents which enhanced mu and kappa binding affinity in the benzomorphan series had a similar effect in the benzofuropyridine series described in this manuscript. An overlap hypothesis topologically connecting the benzomorphan nucleus to the cis-1,2,3,4,4a,9a-hexahydrobenzofuro[2,3-c]pyridine nucleus is also presented. PMID- 2549248 TI - Patulin-induced ion flux in cultured renal cells and reversal by dithiothreitol and glutathione: a scanning electron microscopy (SEM) X-ray microanalysis study. AB - Patulin (PAT), a compound produced by certain species of Aspergillus, Penicillium, and Byssochlamys, is frequently found associated with agricultural commodities. PAT has many effects on membrane function, including the inhibition of the isolated Na+-K+ ATPase. In this study, a scanning electron microscope equipped with an energy dispersive spectroscopy X-ray microanalysis system was used to examine individual cultured renal epithelial cells (LLC-PK1) in order to determine the effects of PAT on the relative intracellular ion concentrations. The estimated EC50 (60 min) for both sodium influx and potassium efflux was between 10 and 50 microns for ouabain. For PAT, the EC50 (60 min) was 250 microns for sodium influx and 100 microns for potassium efflux. However, 1 mM patulin at 240 min caused complete reversal of the sodium and potassium content of cells, and 1 mM ouabain at 240 min did not. The effect of patulin on sodium and potassium flux was both concentration and time dependent and was reversed by dithiothreitol and glutathione. PAT (250 microM) but not ouabain (250 microM) induced massive blebbing of LLC-PK1 cells. Thus, the interaction of PAT with cellular membranes involves both alterations in the regulation of intracellular ion content and the cytoskeleton. We hypothesize that patulin alters intracellular ion content via Na+-K+ ATPase and non-Na+-K+ ATPase mechanisms. PMID- 2549249 TI - Immunodetection of bluetongue virus and epizootic hemorrhagic disease virus in Culicoides variipennis (Diptera: Ceratopogonidae). AB - The avidin-biotin complex immunoperoxidase system has been used with nitrocellulose membranes to detect bluetongue virus serotypes 10 (BTV-10) and 11 (BTV-11) and epizootic hemorrhagic disease virus (EHDV) serotype 1 (EHDV-1) in individual specimens of Culicoides variipennis (Coquillett). The cross reactivity between BTV-10 and BTV-11 indicates that polyclonal antisera cannot be used to distinguish between flies infected with BTV-10 and BTV-11; however, the procedure can be used to discriminate flies infected with BTV-10 or BTV-11 from flies infected with EHDV-1. Because the test is rapid and sensitive and does not require expensive equipment, it could be used in the field to determine the relative numbers of flies infected with BTV or EHDV. PMID- 2549250 TI - Amine and carboxylate spin probe permeability in red cells. AB - Permeabilities for a homologous series of amine and carboxylate nitroxide spin probes were measured in human red blood cells by an electron paramagnetic resonance (EPR) method. Permeabilities determined in this study are much lower than would be predicted for a sheet of bulk hydrocarbon and the polarity of the rate-limiting region is shown to be greater than bulk hydrocarbon. This suggests that the rate-limiting region for permeation of these nonelectrolytes is somewhere in the membrane periphery rather than in the center of the membrane. The red cell membrane does not discriminate between these probes on the basis of molecular volume, as might be predicted by a simple free-volume theory of membrane permeation. PMID- 2549251 TI - Amine spin probe permeability in sonicated liposomes. AB - Permeabilities for an homologous series of amine nitroxide spin probes were measured in liposomes of varying composition by an electron paramagnetic resonance (EPR) method. Results show that the rate-limiting step in permeation is not adsorption/desorption at the aqueous/membrane interface for two probes in phosphatidylcholine/phosphatidic acid liposomes and for one probe in phosphatidylcholine/cholesterol/phosphatidic acid liposomes. Accordingly, we interpret observed selectivity patterns for the entire series of probes in liposomes and red cells in terms of the properties of the bilayer interior. Results are inconsistent with simple applications of either free volume or hydrocarbon sheet models of nonelectrolyte permeation. In the former case, it was found that liposomes do not select against these probes on the basis of molecular volume. In the latter case, probe permeabilities are all much lower than would be predicted for a sheet of bulk hydrocarbon and the polarity of the rate-limiting region is shown to be greater than bulk hydrocarbon. Together with the results of previous studies of spin-labeled solutes in membranes, as well as studies of lipid dynamics in membranes, these latter results suggest that the rate-limiting region in nonelectrolyte permeation is not in the center of the bilayer, but in the relatively ordered acyl chain segments near the glycerol backbone. PMID- 2549252 TI - Ca2+ dependency of Na+ transport by rabbit renal brush border membrane. AB - Intracellular Ca2+ has been suggested to play an important role in the regulation of epithelial Na+ transport. Previous studies showed that preincubation of toad urinary bladder, a tight epithelium, in Ca2+-free medium enhanced Na+ uptake by the subsequently isolated apical membrane vesicles, suggesting the downregulation of Na+ entry across the apical membrane by intracellular Ca2+. In the present study, we have examined the effect of Ca2+-free preincubation on apical membrane Na+ transport in a leaky epithelium, i.e., brush border membrane (BBM) of rabbit renal proximal tubule. In contrast to toad urinary bladder, it was found that BBM vesicles derived from proximal tubules incubated in 1 mM Ca2+ medium exhibited higher Na+ uptake than those derived from proximal tubules incubated in Ca2+-free EGTA medium. Such effect of Ca2+ in the preincubation medium was temperature dependent and could not be replaced by another divalent cation. Ba2+ (1 mM). Ca2+ in the preincubation medium did not affect Na+-dependent BBM glucose uptake, and its effect on BBM Na+ uptake was pH gradient dependent and amiloride (10(-3) M) sensitive, suggesting the involvement of Na+/H+ antiport system. Addition of verapamil (10(-4) M) to 1 mM Ca2+ preincubation medium abolished while ionomycin (10(-6) M) potentiated the effect of Ca2+ in the preincubation medium is likely to be mediated by Ca2+-dependent cellular pathways and not due to a direct effect of extracellular Ca2+ on BBM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549253 TI - Comparative study of K channel behavior in beta cell lines with different secretory responses to glucose. AB - The patch-clamp technique was used to identify and investigate two K channels in the cell membrane of the HIT cell, an insulin secreting cell line with glucose sensitive secretion. Channel characteristics were compared with those of glucose modulated K channels in the RINm5F cell, an insulin secreting cell line in which secretion is largely glucose insensitive. A 65.7 pS channel, identified with the ATP-sensitive K channel was seen in HIT cell-attached patches. Channel activity was dose-dependently inhibited by glucose, by 50 and 100% at 450 microM and 8 mM glucose, respectively, similar to the values previously reported for RIN cells. In inside-out patches channel activity was 50% inhibited by 56 microM ATP and completely blocked between 500 microM and 1 mM, again, similar to the values reported for RIN cells. As in RIN cells a second, considerably larger (184 pS), K channel was glucose sensitive; the glucose sensitivity was similar to that in RIN cells with 50 and 100% channel inhibition at 7.5 and 25 mM, respectively. After patch excision the mean channel conductance increased from 184 to 215 pS. Under these conditions activity was strongly calcium dependent in the range pCa 5-7, identifying this as a calcium- and voltage-dependent K (K(Ca, V] channel; the calcium sensitivity was similar to that of the adult rat beta cell K(Ca, V) channel. In inside-out RIN cell patches, the large K channel was less abundant but displayed a similar conductance (223 pS). However, its calcium sensitivity was more than 10 times lower than in HIT cells, similar to that of the K(Ca, V) channel in the neonatal rat beta cell, which also displays a reduced secretory response to glucose. Based on these observations, it is proposed that the low calcium sensitivity of the K(Ca, V) channel may be causally associated with secretory deficiency in RIN cells and the immature secretory response of the neonatal beta cell. PMID- 2549254 TI - Function of DNA topoisomerases as replication swivels in Saccharomyces cerevisiae. AB - We have examined the roles of eukaryotic DNA topoisomerases I and II in DNA replication by the use of a set of four isogenic strains of Saccharomyces cerevisiae that are TOP1+ TOP2+, TOP1+ top2 ts, delta top1 TOP2+, and delta top1 top2 ts. Cells synchronized by treatment with the alpha-mating factor, or by cycles of feeding and starvation, were released from cell-cycle arrest, and the size distribution of DNA chains that were synthesized after the cells reentered the S-phase was determined as a function of time. The results indicate that synthesis of short DNA chains several thousand nucleotides in length can initiate in the absence of both topoisomerases, but their further elongation requires at least one of the two topoisomerases. Inactivation of DNA topoisomerase II does not alter significantly the time dependence of the patterns of nascent DNA chain synthesis, which is consistent with the notion that the requirement of this enzyme for viability is due to its essential role during mitosis, when pairs of intertwined newly replicated chromosomes are being segregated. The absence of DNA topoisomerase I leads to a temporary delay in the extension of the short DNA chains; this delay in chain elongation is also reflected in the rate of total DNA synthesis in the delta top1 mutant during the early S-phase. Thus, in wild-type cells, DNA topoisomerase I is probably the major replication swivel. The patterns of DNA synthesis in asynchronously grown delta top1 top2 ts cells at permissive and non-permissive temperatures are also consistent with the above conclusions. PMID- 2549256 TI - An approach to study the evolution of the Drosophila 5S ribosomal genes using P element transformation. AB - The P-element-mediated gene transfer system was used to introduce Drosophila teissieri 5S genes into the Drosophila melanogaster genome. Eight transformed D. melanogaster strains that carry D. teissieri 5S mini-clusters consisting of 9-21 adjacent 5S units were characterized. No genetic exchanges between D. melanogaster and D. teissieri 5S clusters were detected over a 2-year survey of the eight strains. The occurrence of small rearrangements within the D. melanogaster 5S cluster was demonstrated in one of the transformed strains. PMID- 2549257 TI - Classical plant taxonomic ambiguities extend to the molecular level. AB - The molecular evolution of cytochrome c from angiosperms is compared to that from vertebrates. On the basis of a cladistic analysis from 26 plant species, compared to that from 27 vertebrate species, we find that although the vertebrate sequences yield reasonably well-defined minimal trees that are congruent with the biological tree, the plant sequences yield multiple minimal trees that are not only highly incongruent with each other, but none of which is congruent with any reasonably biological tree. That is, the plant sequence set is much more homoplastic than that of the animal. However, as judged by the relative rate test, the extent of divergence, and degree of functional constraint, cytochrome c evolution in plants does not appear to differ from that of vertebrates. PMID- 2549255 TI - Evolution of chromosome bands: molecular ecology of noncoding DNA. AB - Giemsa dark bands, G-bands, are a derived chromatin character that evolved along the chromosomes of early chordates. They are facultative heterochromatin reflecting acquisition of a late replication mechanism to repress tissue-specific genes. Subsequently, R-bands, the primitive chromatin state, became directionally GC rich as evidenced by Q-banding of mammalian and avian chromosomes. Contrary to predictions from the neutral mutation theory, noncoding DNA is positionally constrained along the banding pattern with short interspersed repeats in R-bands and long interspersed repeats in G-bands. Chromosomes seem dynamically stable: the banding pattern and gene arrangement along several human and murine autosomes has remained constant for 100 million years, whereas much of the noncoding DNA, especially retroposons, has changed. Several coding sequence attributes and probably mutation rates are determined more by where a gene lives than by what it does. R-band exons in homeotherms but not G-band exons have directionally acquired GC-rich wobble bases and the corresponding codon usage: CpG islands in mammals are specific to R-band exons, exons not facultatively heterochromatinized, and are independent of the tissue expression pattern of the gene. The dynamic organization of noncoding DNA suggests a feedback loop that could influence codon usage and stabilize the chromosome's chromatin pattern: DNA sequences determine affinities of----proteins that together form----a chromatin that modulates----rate constants for DNA modification that determine----DNA sequences. Theories of hierarchical selection and molecular ecology show how selection can act on Darwinian units of noncoding DNA at the genome level thus creating positionally constrained DNA and contributing minimal genetic load at the individual level. PMID- 2549258 TI - Two new members of the OmpR superfamily detected by homology to a sensor-binding core domain. AB - The OmpR superfamily includes proteins that act as transcriptional regulators of operons that respond to environmental stimuli. A homologous domain near the N terminus, termed a sensor-binding core domain, is thought to play a role in recognition of a signal transduction protein. We have identified two previously unrecognized members of this regulator family of proteins: a 23.8-kd protein transcribed from the uvrC transcription unit and the PgtA gene product, which is a phosphoglycerate transport regulatory protein. The sensor-binding core domain is also present in four proteins that regulate bacterial sporulation and chemotaxis. The 23.8-kd protein also has sequence similarity to elongation factor Tu and two regulatory proteins: HtpR, the heat-shock regulatory protein, and TraJ, a regulator of expression of genes involved in conjugation. There is a 77 amino acid region near the C-terminus of the 23.8-kd protein that has 30% similarity with a 28.1-kd protein coded for by an open reading frame 5' to the reading frame of the 23.8-kd protein in the uvrC transcription unit. Genetic distance analysis of amino acid sequences of proteins with a sensor-binding core domain suggests that the 23.8-kd protein and the chemotaxis regulatory proteins are distantly related to the other regulatory proteins in the OmpR superfamily. PMID- 2549259 TI - Mu transposable elements are structurally diverse and distributed throughout the genus Zea. AB - The Robertson's Mutator stock of maize exhibits a high mutation rate due to the transposition of the Mu family of transposable elements. All characterized Mu elements contain similar approximately 200-bp terminal inverted repeats, yet the internal sequences of the elements may be completely unrelated. Non-Mutator stocks of maize have a 20-100-fold lower mutation rate relative to Mutator stocks, yet they contain multiple sequences that hybridize to the Mu terminal inverted repeats. Most of these sequences do not cohybridize to internal regions of previously cloned Mu elements. We have cloned two such sequences from the maize line B37, a non-Mutator inbred line. These sequences, termed Mu4 and Mu5, have an organization characteristic of transposable elements and possess approximately 200-bp Mu terminal inverted repeats that flank internal DNA, which is unrelated to other cloned Mu elements. Mu4 and Mu5 are both flanked by 9-bp direct repeats as has been observed for other Mu elements. However, we have no direct evidence that they have recently transposed because they have not been found in known genes. Although the internal regions of Mu4 and Mu5 are not related by sequence similarity, both elements share an unusual structural feature: the terminal inverted repeats extend more than 100 bp internally from Mu similar termini. The distribution of these elements in maize lines and related species suggests that Mu elements are an ancient component of the maize genome. Moreover, the structure of the Mu termini and the fact that Mu termini are found flanking different internal sequences leads us to speculate that Mu termini once may have been capable of transposing as independent entities. PMID- 2549260 TI - Familial adenomatous polyposis: a nutritional intervention trial. PMID- 2549261 TI - Effect of wheat fiber and vitamins C and E on rectal polyps in patients with familial adenomatous polyposis. AB - Over a 4-year period in a chemoprevention trial on large bowel neoplasia, 58 patients with familial adenomatous polyposis were treated with 4 g of ascorbic acid (vitamin C)/day plus 400 mg of alpha-tocopherol (vitamin E)/day alone or with a grain fiber supplement (22.5 g/day). In this randomized, double-blind, placebo-controlled study, we determined the effects of these supplements on rectal polyps in these patients. Analysis by intent to treat suggested that the high-fiber supplement had a limited effect. Analysis adjusted for patient compliance showed a stronger benefit from the high-fiber supplement during the middle 2 years of the trial. The results provide evidence for inhibition of benign large bowel neoplasia by grain fiber supplements in excess of 11 g/day in this study population. The findings are consistent with the hypothesis that dietary grain fiber and total dietary fat act as competing variables in the genesis of large bowel neoplasia. PMID- 2549262 TI - Induction of renal tumors in cynomolgus monkeys (Macaca fascicularis) by prenatal exposure to 1,2-dimethylhydrazine. AB - Nephroblastomas were induced at an early age in two of six cynomolgus monkeys by prenatal exposure to 1,2-dimethylhydrazine. In one, the tumor of the right kidney had metastasized into the lower lobes of the lungs. By their histologic structure, development, and clinical course, the induced nephroblastomas were analogous to the Wilms' tumors often found in children. PMID- 2549263 TI - TIS gene expression in cultured rat astrocytes: induction by mitogens and stellation agents. AB - The expression of a number of TIS genes (Lim et al.: Oncogene 1:263-270, 1987) was examined in secondary cultures of rat neocortical astrocytes treated with mitogens and stellation agents, to study the early nuclear events which accompany the induction of glial proliferation and/or differentiation. Tetradecanoyl phorbol acetate (TPA), epidermal growth factor, and fibroblast growth factor, three mitogens for astrocytes, stimulated marked, rapid, and transient increases in TIS gene mRNAS. TIS10, which is not expressed in rat PC12 pheochromocytoma cells, could be induced by these mitogens in rat astrocytes. Dibutyryl cyclic adenosine monophosphate and forskolin, which induce rapid stellation in astrocytes, and ganglioside GM1, a potent mitogen as well as an antagonist of the induction and maintenance of stellation, all induced TIS gene expression. Thus, a broad range of agents which elicit both proliferative and differentiation responses in astrocytes are capable of inducing a family of genes that may play a role in the early events of signal transduction. PMID- 2549264 TI - GABAA and GABAB receptors are functionally active in the regulation of catecholamine secretion by bovine chromaffin cells. AB - GABA stimulates the basal catecholamine release from adrenal bovine chromaffin cells in a calcium-dependent manner. This release represents about 70% of that obtained by similar doses of nicotine under similar experimental conditions. This effect is mediated by GABAA receptor sites present in chromaffin cells, since it was mimicked by muscimol and reversed by bicuculline. In addition, GABA, through its GABAA receptors, increases the catecholamine release evoked by submaximal doses of nicotine, but it has no effect on nicotine-evoked secretion of catecholamines when nicotine was given at maximal doses. These results seem to indicate that both nicotine and GABA release catecholamines from the same intracellular pool. In contrast, baclofen, a GABAB receptor agonist, depressed both basal and nicotine-evoked catecholamine release; this result indicates that in addition to GABAA control of catecholamine secretion by chromaffin cells, there is a GABAB control of this function. These results support the existence of a dual regulation of catecholamine secretion by both the GABAA and GABAB receptors in a similar way as that proposed for muscarinic and nicotinic cholinergic receptors. PMID- 2549265 TI - UDP galactose:ceramide galactosyltransferase, CDP choline:1,2-diacyl-sn-glycerol phosphocholine transferase, and microsomal reductases in major regions of the developing rat brain in nutritional stress. AB - The influence of nutritional inadequacy during the active growth phase of the developing brain, coinciding with myelinogenesis, was examined in rat pups. Developmental profiles of enzyme activities involved in biosynthesis of myelin membrane lipids, and those of associated pathways which generate precursor compounds for such biosynthetic reactions, were followed as functions of age in major anatomical regions of the brain. It was observed that while galactosyltransferase and choline phosphotransferase activities were significantly diminished, the microsomal cytochrome reductases, which contribute to the process of fatty acid elongation and desaturation, were also lowered. An attempt at adaptative increase of enzyme activity, in response to the stress, was apparent in the case of glycerol-3-phosphate dehydrogenase, which could possibly explain, in part, the lower susceptibility of phospholipid biosynthesis to impairments induced by nutritional insufficiency. PMID- 2549266 TI - Effect of nerve growth factor on the elongation of neurites from axotomized rat embryonic septal-basal forebrain neurons: an in vitro analysis. AB - The administration of nerve growth factor (NGF) into the brain of a fornix fimbria lesioned rat can rescue many cholinergic, septal-basal forebrain (SBF) neurons from imminent cell death. Unfortunately, it is unclear if NGF can stimulate regenerative growth from axotomized, SBF neurons. In the present study, we used an in vitro model system to determine if NGF could affect neurite outgrowth from nonaxotomized and/or axotomized, embryonic SBF neurons. Axotomized neurons were obtained by severing the neuritic fields surrounding embryonic day (E) 15 SBF explants maintained in primary culture. Acetylcholinesterase (AChE) histochemistry was used to assess the effects of NGF on cholinergic neurites. We report that 1) neurite outgrowth on type I collagen from E15 SBF neurons in primary culture (nonaxotomized neurons) was not affected by NGF. 2) NGF enhanced the outgrowth (regeneration) of axotomized, SBF neurons on a collagen substratum; however, neurons had to be treated with NGF both before and after axotomy to stimulate regeneration effectively. Application of NGF either before or after axotomy did not enhance regenerative neurite outgrowth. 3) SBF neurons had to be axotomized for NGF to facilitate neurite outgrowth. This is supported by the observation that SBF explants, initially maintained in NGF-supplemented medium in suspension culture, did not demonstrate enhanced neurite outgrowth in the presence of NGF when plated onto a substratum. 4) The regenerative growth of AChE negative, as well as AChE-positive, neurites was facilitated by NGF treatment. In addition to data concerning neurite outgrowth, we also found that the NGF receptor, as recognized by the antibody 192-IgG, expands its distribution as time in culture progresses; i.e., staining, originally confined to cell bodies and proximal processes within the explant, later included neurites that emanated from the explant. Thus, our results demonstrate that NGF can stimulate regenerative growth from axotomized, but not nonaxotomized, embryonic SBF neurons. We hypothesize that, given the appropriate substratum for axon elongation in vivo, NGF can stimulate the regeneration of SBF neurons in the injured adult brain. PMID- 2549267 TI - Aldosterone secretion following non-hypotensive hemorrhage is potentiated by prior blood loss. AB - Aldosterone is a major regulator of fluid and electrolyte balance after hemorrhage and is released from the adrenal cortex by the action of adrenocorticotropin (ACTH) and angiotensin II (AII). Past work has shown that the hemorrhage-induced release of ACTH and cortisol is potentiated by prior hemorrhage. We therefore studied the response of adrenal aldosterone secretion to repeated hemorrhage and its control by ACTH and AII. Six awake dogs with chronic lumboadrenal vein catheters were bled 10% of measured blood volume (H1) with reinfusion at 30 minutes. The hemorrhage was repeated 5 hours later (H2). Adrenal presentation rates for AII (AII-PR) and ACTH (ACTH-PR) were calculated for each sample. Control hormonal and hemodynamic parameters before each hemorrhage were not different; hemodynamic responses to H1 and H2 did not differ. Aldosterone secretion increased significantly after each hemorrhage. The increase in aldosterone secretion after H1 was associated with an early increase in AII-PR and late increase in ACTH-PR. Aldosterone secretion following H2 was greater than that following H1 and was associated with early and larger responses of AII-PR and ACTH-PR. Aldosterone secretion following H1 correlated with the AII-PR (r = 0.75; p less than 0.001), but not with the ACTH-PR. In contrast, aldosterone secretion following H2 correlated with both the AII-PR (r = 0.54; p less than 0.01) and ACTH-PR (r = 0.71; p less than 0.001) and multiple regression analysis showed a highly significant relation with both AII and ACTH (r = 0.81; p less than 0.001). The data suggest that aldosterone secretion after initial small hemorrhage occurs as a result of increased AII, whereas both AII and ACTH may contribute to the larger aldosterone secretory response to H2. Since major trauma commonly involves at least two insults separated in time (e.g., injury followed by surgery), potentiated responses of aldosterone and other pituitary-adrenal hormones (ACTH, vasopressin, and cortisol) may have important implications for the control of fluid and electrolyte balance and metabolism in injured patients. PMID- 2549268 TI - Interpretation of urine quantitative 11-nor-delta-9 tetrahydrocannabinol-9 carboxylic acid to determine abstinence from marijuana smoking. AB - Verification of abstinence from cannabinoid use after initial identification requires documentation of falling quantitative levels of urinary 11-nor-delta-9 tetrahydrocannabinol-9-carboxylic acid (THC-COOH) over an extended time period. We present a case in which normalization of quantitative urinary THC-COOH results to urinary creatinine is required to correctly interpret a series of quantitative levels and confirm abstinence from marijuana smoking. PMID- 2549269 TI - [The role of ADP-cyclic AMP system in horseshoe crab amebocytes aggregation and gel formation]. AB - A study of the effects of ADP and cyclic AMP related drugs on gel formation induced by endotoxin and amebocytes aggregation was conducted. The gel formation induced by endotoxin could be inhibited by ADP, cyclic AMP, ATP and aminophylline. Conversely, Phosphodiesterase enhanced gelation rate. A calmodulin antagonist, trifluoperazine, also inhibited endotoxin-induced gel formation, but this agent was able to induce potentiation in amebocytes aggregation. These results suggest that the cyclic AMP phosphodiesterase system may play an important role in endotoxin-induced gel formation, and it was concluded that, in the presence of both Ca++ and endotoxin, the calmodulin-like protein will activate the cyclic AMP phosphodiesterase system and decrease the concentration of cyclic AMP, thus promoting gel formation. PMID- 2549270 TI - One of 8 women with bladder neoplasia may have concomitant gastrointestinal pathology, including cancer. AB - A study of the pathology records during 6 years for 160 women with bladder neoplasia and 150 with malignant melanoma revealed differences in patient experience in regard to the incidence of surgical biopsies. Biopsy for additional skin lesions was common in melanoma patients, who produced few gastrointestinal lesions. In contrast, 19 of the bladder cancer patients produced 26 gastrointestinal lesions, including 5 carcinomas. The latter incidence is higher than expected in our population. This difference between the 2 sets of patients in regard to skin and gastrointestinal lesions is statistically significant. Few gynecological lesions were noted in either group but a common factor was human papillomavirus infection, as indicated by the presence of koilocytosis. The latter also was present in 17 per cent of 41 otherwise negative smears from patients with bladder neoplasia, that is considerably more than expected (less than 1 per cent), and is similar to that reported previously in patients with melanoma. PMID- 2549271 TI - The urological manifestations of the acquired immunodeficiency syndrome. AB - Between 1984 and March 1987, 120 patients with either the acquired immunodeficiency syndrome or its related complex seen at our hospital were studied retrospectively for urological signs and/or symptoms. Autopsy findings also were reviewed. Of the patients 84 per cent had no complaints referable to the urinary system, 2 per cent had gross hematuria (all with a negative diagnostic evaluation) and 14 per cent had urinary infections. We conclude that only a small percentage of patients with the acquired immunodeficiency syndrome suffer from significant urological manifestations and that a full urological evaluation of such patients generally is not warranted. If the patient presents with gross hematuria excretory urography should be performed if there is no infectious etiology, and cytoscopy should be performed only if the hematuria is life-threatening or prolonged and possibly to confirm significant urographic findings. PMID- 2549272 TI - 99mtechnetium-dimercapto-succinic acid renal scanning and excretory urography in diagnosis of renal scars in children. AB - We compared the ability of excretory urography (without tomography) and 99mtechnetium-dimercapto-succinic acid renal scanning to detect renal scars in 32 children with primary vesicoureteral reflux. These children did not have hydronephrosis, renal failure or urinary tract obstruction. In all cases both studies were conducted within a 10-month period. The findings from both modalities were in agreement for 51 of the 64 renal units evaluated (80 per cent). Evaluation of the excretory urogram indicated 6 cases of diffuse and 2 of focal scarring that were not detected by evaluation of the renal scan. The sensitivity of excretory urography to detect renal scars was 84 per cent and the specificity was 83 per cent. The 99mtechnetium-dimercapto-succinic acid renal scan showed 5 cases of focal renal scarring not detected by excretory urography. The sensitivity of the renal scan to detect renal scars was 77 per cent and the specificity was 75 per cent. We conclude that neither study alone could effectively replace the other for the detection of renal scars, and recommend that both be included in the initial evaluation and followup of patients with renal scars. PMID- 2549273 TI - A maternal functioning adrenocortical adenoma causing fetal female pseudohermaphroditism. AB - Fetal virilization induced by a maternal adrenal tumor is extremely rare. We report a case of a maternal adrenocortical tumor with Cushing's syndrome causing fetal virilization. PMID- 2549274 TI - Successful management of small cell carcinoma of the bladder with cisplatin and etoposide. AB - A 63-year-old white man with metastatic small cell carcinoma of the bladder attained a complete remission with a combination of etoposide and cisplatin chemotherapy, which has lasted for more than 2 years. PMID- 2549276 TI - From the Centers for Disease Control. Progress toward eradicating poliomyelitis from the Americas. PMID- 2549275 TI - Abnormalities of responses to autonomic stimulation in the mouse urinary bladder associated with Semliki Forest virus-induced demyelination. AB - The responses to autonomic stimulation of the urinary bladder were studied in mice infected with Semliki Forest Virus, which has been proposed as an animal model for multiple sclerosis. Mice infected with Semliki Forest Virus showed a decreased body weight when compared with control animals, although their bladders were proportionately heavier. The pharmacological results indicated a selective change in purinergic transmission in the bladders of mice infected with Semliki Forest Virus, while cholinergic transmission remained unchanged. This was demonstrated by a significant increase in the contractile response to beta,gamma methylene ATP accompanied by an increase in the proportion of the nerve-mediated response blocked by alpha,beta-methylene ATP. In contrast, the contractile response to acetylcholine and the atropine-sensitive component of the neurogenic response were not significantly different between the two groups of animals. Active length-tension curves were also unchanged in the mice infected with Semliki Forest Virus when compared with controls. The results are discussed in relation to those obtained from models of urinary bladder dysfunction associated with hypertrophy; whether the alteration of atropine-resistant (purinergic) function reported here is a general secondary feature of this type of bladder dysfunction or a change specific to the Semliki Forest Virus model of multiple sclerosis is not resolved. This study provides evidence that peripheral changes as well as central nervous system demyelination occur in the Semliki Forest Virus infected mouse. PMID- 2549277 TI - From the Centers of Disease Control. Enterovirus surveillance--United States, 1989. PMID- 2549278 TI - Management of infectious waste by US hospitals. AB - In July 1987 and January 1988, forty-six percent (441/955) of randomly selected US hospitals responded to a questionnaire intended to identify their waste disposal practices. Survey responses were received from hospitals in 48 states. United States hospitals generated a median of 6.93 kg of hospital waste per patient per day and infectious waste made up 15% of the total hospital waste. Most hospitals (greater than 90%) considered blood, microbiology, "sharps," communicable disease isolation, pathology, autopsy, and contaminated animal carcass waste as infectious. Other sources of hospital waste that were commonly (greater than 80%) designated infectious were surgical, dialysis, and miscellaneous laboratory waste. The infectious waste was normally (80%) treated via incineration or steam sterilization before disposal, whereas noninfectious waste was discarded directly in a sanitary landfill. Eight-two percent of these US hospitals are discarding blood, microbiology, sharps, pathology, and contaminated animal carcass waste in accordance with the Centers for Disease Control's recommendations, while the compliance rate for the Environmental Protection Agency's recommendations (excluding optional waste) is 75%. No hospital could identify an infection problem (excluding needle-stick injuries) that was attributable to the disposal of infectious waste. While the management of infectious waste by US hospitals is generally consistent with the Centers for Disease Control's guidelines, many hospitals employ overly inclusive definitions of infectious waste. PMID- 2549279 TI - [A comparative study of the biological behavior of gastric cancer from the view of the amount of connective tissue in the cancerous tissue--a correlation between the amount of connective tissue and the metastatic pattern]. AB - Differences between advanced gastric cancer of the medullary type (med.: 138 cases) and the scirrhous type (sci: 164 cases) with regard to the amount of connective tissue that was cancerous have been studied clinicopathologically. A correlation was found between med and liver metastasis, and sci and peritoneal dissemination in metastatic types, whereas no correlation was observed in other factors except between med and a localized type, sci and infiltrating type in gross type. The study of tumorigenicity in nude mice showed remarkable difference between med (86.7%) and sci. (0%). These results indicate that there is definite relationship between the amount of connective tissue and the biological behavior of a gastric cancer. PMID- 2549280 TI - [Hepatocellular carcinoma treated with a curative segmentectomy five years after complete remission of acute myeloblastic leukemia]. AB - Acute myeloblastic leukemia (AML) was diagnosed in a 54-year-old male who had been known to carry a chronic hepatitis B surface antigen (HBsAg) from June, 1983. Prompt remission was achieved with combination chemotherapy of BHAC-DMP. Follow-up maintenance and an intensification of this chemotherapy had been given for five years. He was readmitted to our hospital in March, 1988 because a mass was detected in the right lobe of the liver by ultrasonography. His serum alpha fetoprotein (AFP) level was found to be 180.1 ng/ml, and was diagnosed as having a hepatocellular carcinoma though there was no evidence of liver cirrhosis. A curative right hepatectomy was performed in May, 1988 after transcatheter arterial embolization and portal embolization. After resection of the tumor, the AFP level decreased to 10.7 ng/ml and no HbsAg was detected in the serum. PMID- 2549281 TI - [A carcinoma of the right breast arising after a mastectomy in primary malignant lymphoma of the left breast]. AB - In 1987, a 51-year-old woman visited our hospital, having noticed a painless 1.5 x 1.5 cm tumor of the left breast. On biopsy, it was pathologically diagnosed as being a primary malignant lymphoma of the breast. Thus we performed a modified radical mastectomy (Patey's method), in combination with chemotherapy (Cyclophosphamide, Vincristine, Prednisolone). Two years after the operation, a tumor of the right breast, 1.0 x 1.0 cm in size, was noticed. At this time, since an excised specimen was pathologically determined as being a scirrhous carcinoma, we performed a modified radical mastectomy (Auchincloss' method), followed by chemotherapy (Tegafur). PMID- 2549282 TI - [Gastric acid secretions and serum gastrin levels in patients with mucosal and submucosal gastric cancers]. AB - Gastric acid secretions and serum gastrin levels have been examined in 128 patients with early gastric cancer and in 98 gastric ulcer patients. Gastric cancer patients were found to have lower acid secretions than did gastric ulcer patients, and those with elevated types of a differentiated adenocarcinoma had lower acid secretions than did those with depressed types of an undifferentiated adenocarcinoma. Gastric acid secretions in patients with both a gastric ulcer and cancer were found to decrease with aging. However, the serum gastrin levels were found to be decreased in patients with a gastric ulcer and to be increased in patients with a gastric cancer. Incidences of a differentiated adenocarcinoma increased with aging. From these observations, it has been speculated that the carcinogenesis of a differentiated adenocarcinoma may be related to increasing endogenous gastrin levels and decreasing gastric acid secretions. These results suggest that a continuous check of the serum gastrin levels might be a good marker for cancer detection and that gastrin antibodies might be useful for treatment. PMID- 2549284 TI - [Parvovirus-induced aplastic crisis in a child with hereditary spherocytosis: studies on the mechanisms of hemopoietic suppression]. AB - A 4 4/12-year-old girl with hereditary spherocytosis (HS) who presented with an aplastic crisis during a human parvovirus (HPV) B19 infection is reported. IgM and IgG antibodies to the HPVB19 and HPV19 DNA were detected. Each of Leu 7+, Leu 11+ and HLA-DR+ cells increased. OKT4/OKT8 ratio decreased to 0.71. In order to investigate the mechanism of aplastic crisis, we used an in vitro culture technique for erythroid and granulocyte-macrophage progenitor cells (BFU-E, CFU-E and CFU-C). The patient's HPV19 DNA-containing aplastic phase serum inhibited the formation of BFU-E, CFU-E and CFU-C. After removal of the adherent cells from the aplastic phase bone marrow, the numbers of BFU-E significantly increased. These results suggest that aplastic crisis of the patient with HS was caused by HPVB19, and that monocytes-macrophages and NK cells played an important role in the pathogenesis of aplastic crisis. PMID- 2549283 TI - [An autopsy case of AIDS with disseminated cytomegalovirus infection and neurological disturbance]. AB - We report a case of acquired immunodeficiency syndrome (AIDS) complicated by disseminated CMV infection and neurological disturbance. A 21 years old male with hemophilia A was diagnosed as having AIDS in Feb. 1986 because of interstitial pneumonia and esophageal candidiasis. Since Jan. 1987 he had complained of hypesthesia in the legs. On Mar. 14 he was admitted due to diarrhea. The laboratory data revealed that WBC was 4,000/microliters including 29% of lymphocytes, 1.6% of OKT4+-, 71.6% of OKT8+-lymphocytes, T4/T8 ratio 0.02 and positive HIV antibody and HTLV-1 antibody. After the admission, sensory disturbance exacerbated to complicate paraplegia. He developed acute hepatitis associated with leukopenia, thrombocytopenia, pneumonia and melena, and eventually died on May 29. The autopsy findings disclosed CMV infection in the lungs, colons, and adrenal glands, suggesting that the primary cause of death was adrenal insufficiency. Degeneration of cerebro-spinal nerve cells and peripheral neuritis were thought to result from direct HIV infection to the nervous system. PMID- 2549285 TI - [Extensive bone marrow necrosis associated with carcinomatosis 7 years after operation for gastric cancer]. AB - A case of extensive bone marrow necrosis due to cancer metastasis is reported. A 55-year-old female, who had a history of subtotal gastrectomy for signet ring cell carcinoma of the stomach 7 years ago, was admitted to our hospital with a complaint of lumbago on October 25, 1987. Red blood cell count was 92 X 10(4)/microliters, hemoglobin 2.7 g/dl, hematocrit 8.0%, platelet 6.4 X 10(4)/microliters, and white blood cell count 13,400/microliters with leukoerythroblastosis. Bone marrow aspiration of the sternum, left iliac crest, and bilateral posterior superior iliac supine showed extensive bone marrow necrosis. Serum ALP was increased to 7410IU/l, dominated isozyme of bone type. Hemostatic findings suggested a complication of consumption coagulopathy. Skull, vertebrae, iliac and pelvic bone X-ray showed multiple osteolytic lesions, and irregular isotope uptake was recognized on the bone scintigraphy using 99mTc. Sixth bone marrow examination at the right iliac crest revealed signet ring cell carcinoma metastasis. In spite of detailed examinations, there was no evidence of primary carcinoma, including the remnant of stomach. We speculated that the signet ring cells were originated from the respected gastric cancer. The patient has received anti-cancer chemotherapy with UFT and OK432, and is still alive 9 months after diagnosis. PMID- 2549286 TI - [Targeting cancer chemotherapy used Lipiodol as a carrier]. PMID- 2549287 TI - [Clinico-pathological analysis of anti-early antigen (EA)-IgG antibodies in Epstein-Barr virus infection]. PMID- 2549288 TI - [Late radiation effects of low doses from occupational exposure. Antibodies to cytomegalovirus, Epstein-Barr virus and human T cell lymphotropic virus type 1 in radiological technologists]. PMID- 2549289 TI - [Detection of rotavirus RNA by a dot hybridization using RNA probe]. PMID- 2549290 TI - [The hepatic hemodynamic response to intra-arterial infusion of vasoactive agents -Part 1. Blood flow measurements in dogs]. AB - The common hepatic arterial (CHA), portal venous (PV) and liver tissue (LT) blood flows were measured during continuous infusion of vasoactive agents to the celiac axis under the observation of systemic circulation. The infusions of Angiotensin II (AT) (0.05 microgram/kg/min) and Prostaglandin F2 alpha (PGF) (0.025-0.05 microgram/kg/min) reduced CHA and LT blood flow, but caused no remarkable change of PV blood flow. The infusions of AT (0.5 microgram/kg/min) and PGF (0.5 microgram/kg/min) caused a biphasic response in CHA blood flow: an initial decrease in CHA blood flow was followed by a marked increase in this parameter (p less than 0.05), while PV blood flow decreased, but LT blood flow remained unchanged. The infusion of PGF (1.0 microgram/kg/min) reduced all parameters of CHA, PV and LT blood flows. The infusion of Dibutyryl cyclic AMP (DBcAMP) (2.0 mg/kg/hr) increased all parameters of CHA, PV and LT blood flows. Preinfusion of DBcAMP weakened the primary effects of PGF (1.0 microgram/kg/min) infusion. PMID- 2549291 TI - [Quantification of myocardial infarct size by technetium-99m pyrophosphate single photon emission computed tomography]. AB - Myocardial infarct size in 41 patients with the first attack of acute transmural myocardial infarction (MI) was assessed by technetium-99m pyrophosphate single photon emission computed tomography (99mTcPYP-SPECT). A ratio of the number of voxels of 99mTcPYP uptake into the infarct area to that into the thorax was calculated as a parameter of MI size. The ratio was positively correlated with both peak CPK activity (r = 0.53, p less than 0.005, n = 24) and extent score in 201TI-SPECT (r = 0.70, p less than 0.005, n = 14) significantly in patients with anterior MI but not in patients with inferior MI. There was also significant negative correlation between the ratio and the left ventricular ejection fraction (LVEF) measured by RI angiography in both acute (r = -0.67, p less than 0.005, n = 18) and chronic (r = -0.75, p less than 0.005, n = 25) phases in patients with anterior MI. Recovery in LVEF at chronic phase was noted in patients with small anterior MI but not with large anterior MI. 8 of 14 patients with inferior MI had right ventricular MI, that might have affected evaluation of MI size and resulted in no correlation between variables. It was suggested that 99mTcPYP-SPECT was a useful method to evaluate MI size and to predict prognosis of cardiac function in patients with anterior MI but not in patients with inferior MI. PMID- 2549292 TI - Biphasic accumulation of leukocytes in rat cardiac infarct tissue caused by leukotriene B4 and complement. AB - An initial increase (3-12 hr) in the polymorphonuclear leukocytes (PMN) counts after ligation of the rat left main coronary artery was reduced by 49.4% (at 12 hr) by a 5-lipoxygenase inhibitor, AA-861 (80 mg/kg, p.o., 1 hr before ligation). Depletion of the complement components induced by cobra venoma factor (CVF) (i.v.), given one day before, resulted in significant reduction in the PMN accumulation after 12 hr (by 63.6% at 24 hr). The combined treatment (CVF+AA-861) suppressed the PMN accumulation by 69.7% (24 hr). The infarct size at 48 hr was also reduced by approximately 36% by either AA-861, CVF or combined treatments. PMID- 2549293 TI - Production of tumor necrosis factor by rat mesangial cells in response to bacterial lipopolysaccharide. AB - Tumor necrosis factor (TNF) is a cytokine which is produced by mononuclear phagocytes upon activation by bacterial lipopolysaccharide (LPS) and various other stimuli. In immune-mediated glomerulonephritis, infiltration of glomeruli by monocytes-macrophages is associated with production of TNF. The purpose of the present experiments was to determine whether mesangial cells could also contribute to glomerular TNF synthesis. TNF activity has been determined in the culture medium of rat mesangial cells using a L-929 fibroblast lytic assay. This activity was detectable only when the cells were exposed to LPS (0.1 to 10 micrograms/ml) and for periods longer than one hour. The cytotoxic factor was identified as TNF since: (1) the lytic activity was completely inhibited by an anti-mouse TNF polyclonal antibody and was associated with suppression of lipoprotein lipase activity in adipocytes; (2) its molecular weight (110,000 daltons) corresponded to that observed for murine TNF under non-denaturing conditions; and (3) mRNA encoding TNF was expressed by mesangial cells two hours after addition of LPS. To assess the mechanisms whereby TNF production was regulated, the role of prostaglandin E2 (PGE2) was determined. LPS caused a dose dependent increase of PGE2 synthesis by mesangial cells. Treatment by indomethacin promoted a suppression of PGE2 production together with an increase of TNF synthesis, indicating that PGE2 acted in a negative feedback manner to regulate the production of TNF. Addition of PGE2 (0.1 to 300 nM) or 8-bromo cyclic AMP (0.1 to 100 microM) induced similar dose-dependent reductions of TNF synthesis. Thus the inhibitory effect of PGE2 probably required in part cyclic AMP accumulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549294 TI - Micromolar aluminum levels reduce 3H-thymidine incorporation by cell line UMR 106 01. AB - Aluminum-induced osteomalacia is a frequent complication observed in patients on maintenance hemodialysis. However, it is not known whether there are direct effects of aluminum on osteoblasts, or alternatively, whether the observed changes are due to changes in PTH or other factors. We sought to determine the effect of micromolar levels of aluminum on osteoblasts using a well-defined cell line derived from a 32P induced osteosarcoma of rat, UMR 106-01, which is alkaline-phosphatase positive, responds to PTH, and synthesizes type I collagen. Aluminum exposure was controlled using tissue culture media with [Al ] less than 1 microgram/liter (40 nM), produced by precipitation of aluminum salts at pH 8.5. The effect of defined [Al ], from 20 to 800 micrograms/liter (0.7 to 30 microM), was then determined by adding back aluminum while measuring DNA and protein synthesis. We found that aluminum depressed DNA synthesis, as determined by 3H thymidine incorporation, by 60%, with half maximal effect at 20 micrograms/liter (740 nM) in cells at a density of 20,000/cm2. Alternatively, protein synthesis, as determined by 3H-leucine incorporation, did not decline, and in some cases increased. However, qualitative analysis of matrix proteins produced with and without 800 micrograms/liter (30 mM) [Al ] showed no differences. Direct measurements of cell number and protein synthesis confirmed these findings. Al does not alter the PTH-induced cAMP response of these cells. Thus, aluminum has a direct effect on cell division, and probably on protein synthesis, in this osteoblast-like cell line. These effects occur at levels of aluminum below those commonly contaminating tissue culture media, and thus are seen reproducibly only in media of defined [Al ].(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549295 TI - Hepatoma G2 conditioned medium facilitates early outgrowth of endothelial cells from isolated glomeruli. PMID- 2549296 TI - Prior inhibition of vasoconstrictors normalizes GFR in postobstructed kidneys. AB - The present studies were designed to analyze the potential contribution of angiotensin II and thromboxane A2 to the remarkable decrease in glomerular filtration rate (GFR) and renal plasma flow observed after unilateral release of 24-hour bilateral ureteral obstruction. Pretreatment of the animals with inhibitors of either thromboxane or angiotensin synthesis for 48 hours prior to and during obstruction eliminated the contribution of these vasoconstrictors. Inhibition of these vasoconstrictors during the period of obstruction results in a greater increase in renal plasma flow and GFR than when inhibition was accomplished after release of the obstruction. These data suggest a greater role for these vasoconstrictors in the decrease in GFR that occurs with obstruction. Simultaneous inhibition of thromboxane and angiotensin production normalized GFR of the postobstructed kidney. Administration of atrial peptide after release of obstruction in the different groups of rats resulted in further increases in GFR, urine flow and absolute sodium excretion. It is suggested that atrial peptide participates in the renal hemodynamic changes that occur in the postobstructed kidney. PMID- 2549297 TI - Cushing's syndrome due to ACTH-production of an ovarian carcinoid. AB - The case of a 56-year old woman with severe Cushing's syndrome due to ovarian ACTH-production is described. Both clinical picture and biochemical pattern were consistent with the ectopic ACTH syndrome. ACTH was found by specific immunohistochemical staining in a carcinoid tumor of the patient's right ovary. In contrast, pituitary cells exhibited immunoreactive ACTH to only a minimum extent. PMID- 2549298 TI - [Status of the pituitary-adrenal system in patients with non- specific ulcerative colitis]. AB - The blood content of basal cortisol was studied in 54 patients with a total form of non-specific ulcerous colitis and in 23 of them that of ACTH. A low cortisol level was revealed in 71.4 per cent of the patients, while the level of ACTH remained unchanged. Treatment with prednisolone (1-2 mg/kg) for 6-9 weeks did not deteriorate the functional state of the hypophyseal-adrenal system. A single administration of the total daily dose of prednisolone in the morning protected the adrenal cortex from exhaustion and in some of the patients even caused increased in and normalization of the cortisol level. Rectal administration of hydrocortisone (125 mg), which was included in therapeutic scheme ensured normalization of the blood plasma cortisol level and did not cause decrease in the ACTH level. PMID- 2549299 TI - [Changes in the activities of enzymes of metabolism of DNA precursors in peptic ulcer and cancer of the stomach]. AB - Comparative assessment of the remote results of Hoffmeister-Finsterer gastrectomy with creation of valve anastomoses has demonstrated rare occurrence of postoperative complications owing to anastomoses. The operation is simple as concerns operative techniques and can be recommended as operation of choice in the treatment of peptic ulcer. PMID- 2549300 TI - Comparative analysis of herpes simplex virus type 1 mutants that are defective in fusion function by using restriction endonuclease. AB - Mutants derived from herpes simplex virus type 1 (HSV-1) strains, YH and KOS, that were defective in cell fusion function (r mutants) were examined for DNA structure using a restriction endonuclease and gel electrophoresis. A distinct difference in the restriction patterns was found in some r mutants. One which was derived from strain YH and belonged to complementation group A lost the BamHI cleavage site in the long inverted repeat regions. But the site could not be associated with the gene that was related to the cell fusion event, inasmuch as the other mutants belonging to the same complementation group had a normal BamHI site. The others, which belonged to complementation group G of strain KOS, had a deletion of approximately 2 kb (kilobase-pair) in the BamHI B fragment. A mixed infection was carried out with r410 and r426 which belonged to groups A and G of strain KOS, respectively, and the viruses of parental and recombinant types were isolated from the progeny. The BamHI restriction fragment patterns of the DNAs showed that all recombinants, which carried the cell fusion ability, had the normal BamHI fragment B. Thus, one of the genes associated with cell fusion function must be located in BamHI fragment B. PMID- 2549301 TI - Endocytic pathway of high density lipoprotein via trans-Golgi system in rat resident peritoneal macrophages. AB - Interaction of high density lipoprotein (HDL) with rat resident peritoneal macrophages was investigated by morphological and biochemical approaches. Binding studies at 0 degree C demonstrated saturable binding sites for 125I-HDL on the surface membrane. When cells were incubated with 125I-HDL at 37 degrees C, the cell-associated radioactivity increased with time, but intracellular degradation of HDL did not occur. Rather, the cell-associated 125I-HDL was released intact into the medium. Two morphological probes were employed to visualize the post binding fate of HDL. Horseradish peroxidase and ferritin conjugates of HDL showed specific bindings to the coated pits of the plasma membranes and were internalized and delivered through receptosomes to a trans-Golgi system. They were then resecreted as secretory vesicles from the cells. Parallel experiments with transferrin-horseradish peroxidase conjugates revealed an endocytic pathway identical or very similar to that observed with HDL. These results provide evidence that internalization and subsequent nonlysosomal pathway via the trans Golgi system are involved in receptor-mediated endocytosis of HDL by macrophages. PMID- 2549302 TI - Protein glycosylation and collagen metabolism in normal and diabetic rats. AB - The high concentration of glycosylated extracellular proteins found in diabetics may contribute adversely to wound healing. The degree of collagen glycosylation was correlated with collagenase activity and 5-hydroxyproline (5-OHP) levels in surgical wounds of normal and diabetic animals. Expanded polytetrafluoroethylene (PTFE) wound cylinders were implanted subcutaneously in 10 normal and 10 streptozotocin-induced diabetic rats. Nonenzymatic glycosylation (NEG) of protein was inhibited in half the subjects in each group by daily enteral acetylsalicylic acid (ASA). Wound cylinders were removed on the fifth day and assayed for collagenase activity and 5-OHP levels, and the degree of collagen glycosylation was measured with an assay for 5-hydroxymethylfurfural (5-HMF). Results were compared using Student's two-tailed t test. When compared with controls, diabetic animals had less wound 5-OHP (1.87 +/- 0.01 vs 3.11 +/- 0.03 micrograms/cm PTFE) (P less than 0.0001), higher levels of glycosylated collagen (215 +/- 11 vs 150 +/- 9 micrograms 5-HMF/mg protein) (P less than 0.001), and increased collagenase activity (13.5 +/- 0.6 vs 11.7 +/- 0.5 microgram collagen/mg protein/hr) (P less than 0.001). When ASA was added to the diet of diabetic animals, results in each case were statistically indistinguishable from controls. The collagenase activity of nondiabetic animals receiving ASA was decreased from nondiabetic controls (7.6 +/- 0.5 vs 11.7 +/- 0.5 microgram collagen/mg protein/hr) (P less than 0.001). These data show that ASA inhibits NEG of wound collagen in vivo, and ASA may normalize some abnormalities of wound healing found in diabetic animals. PMID- 2549303 TI - Mineralocorticoid specificity of renal type I receptors: binding and metabolism of corticosterone. AB - In cytosol preparations renal mineralocorticoid receptors have equivalent affinity for aldosterone and corticosterone; in vivo, such receptors are clearly aldosterone-selective. To explore this difference, we have compared the binding of both steroids in kidney slices, high speed supernatants and true cytosols in vitro, in the presence of RU28362 to exclude steroids from Type II glucocorticoid receptors, and/or in the presence of cortisol 17 beta-COOH to exclude corticosterone from transcortin. In addition, we have explored the effect of tissue preparation and cortisol 17 beta-COOH on the level of activity of the renal enzyme 11 beta-OH steroid dehydrogenase. The results of the present studies support the hypothesis that the predominant aldosterone specific-conferring mechanism for renal Type I receptors in vivo in the enzyme 11 beta-OH steroid dehydrogenase, and underline the potential difficulties in interpreting studies which include multiple potential ligands to multiple potential binding sites. PMID- 2549304 TI - Comparison of assays for catechol estrogen synthase activity: product isolation vs radioenzymatic catechol-O-methyltransferase-coupled procedures. AB - Reported values for the activity of enzymes mediating catechol estrogen formation by hamster kidney and liver, measured by catechol-O-methyltransferase-coupled radioenzymatic assay, have been uniformly low and there have been marked discrepancies in values reported from different laboratories. Therefore, we examined the validity of the radioenzymatic assay used in these studies. NADPH dependent estrogen 2- and 4-hydroxylase activity of hamster liver microsomes measured by radioenzymatic assay was comparable to that reported in the literature but at least one order of magnitude lower than that obtained with a direct product isolation assay. Several features of the radioenzymatic assay were identified which, together, contribute to the underestimation of enzyme activity. They include, incomplete protection from oxidative degradation of both the catechol estrogens generated and of the catechol-O-methyltransferase and assay conditions which are suboptimal for O-methylation of the catechol estrogens. We conclude that results obtained using the catechol-O-methyltransferase-based radioenzymatic assay can only be considered valid if a consistent stoichiometric relationship can be demonstrated between the amounts of catechol estrogens and their O-methylated products. PMID- 2549305 TI - Expression of epidermal growth factor receptors in four histologic cell types of lung cancer. AB - Lung cancer tissues from 68 patients were examined for epidermal growth factor (EGF) receptor levels and EGF receptor gene copy numbers. Histologic cell types of these lung cancer tissues included squamous-cell carcinoma (n = 30), adenocarcinoma (n = 28), large-cell carcinoma (n = 4), and small-cell carcinoma (n = 6). Tissues of squamous-cell carcinoma exhibited exceptionally high 125I-EGF binding activity, and those of small-cell carcinoma showed no EGF binding activity. Southern blot hybridization analysis revealed EGF receptor gene amplification in the squamous-cell carcinomas with high EGF binding activity. The EGF receptor levels in squamous-cell carcinomas and adenocarcinomas were compared with their pathological staging grouping and pathological findings, including degree of differentiation, diameter of tumor, and lymph node metastasis. However, unlike previous reports on breast and bladder cancers, there was no obvious correlation between these pathological characteristics and the EGF receptor levels of lung cancer. PMID- 2549306 TI - Neuroendocrine neoplasms of the lung. A clinicopathologic update. AB - One hundred forty-six cases of pulmonary neuroendocrine tumors are assessed according to the classification of Gould and associates and are evaluated for their clinical presentation and subsequent clinical course. Bronchial carcinoids are characteristically found to be central tumors often occurring in comparatively young patients; surgical resection with minimal but clear margins is usually curative. The long-term prognosis is excellent in the majority of patients, although rarely regional nodal and distant metastases develop. Well differentiated neuroendocrine carcinomas are most frequently peripheral tumors. In stage I and II disease, surgical resection alone is curative and patients with locally advanced tumors may have a prolonged disease-free interval. The overall prognosis is less favorable than that of bronchial carcinoids but considerably better than that of small cell neuroendocrine carcinomas, with which they are still at times confused. Intermediate-sized cell neuroendocrine carcinomas are often wrongly categorized as large cell undifferentiated carcinoma. They have a distinctly aggressive clinical course comparable with that of small cell neuroendocrine carcinoma and should be treated similarly. Small cell neuroendocrine carcinomas are aggressive, rapidly disseminating neoplasms. Even in clinical stage I tumors, patients must be considered to have disseminated metastases. The role of surgical therapy in these two latter tumor types is adjuvant to aggressive systemic chemotherapy. PMID- 2549307 TI - Endotoxemia and enhanced generation of oxygen radicals by neutrophils from patients undergoing cardiopulmonary bypass. AB - Plasma endotoxin concentrations and oxidative burst response of peripheral blood polymorphonuclear leukocytes were examined in 12 patients undergoing coronary artery bypass. The measurements were made just before the operation, 5 minutes after removal of the aortic crossclamp, and 24 hours after the operation. Endotoxin was quantitated by a combination of a sensitive Limulus amebocyte lysate assay and rocket immunoelectrophoresis measuring picogram amounts of endotoxin. Peripheral blood neutrophils were purified by a two-step dextran sedimentation and metrizoate sodium Ficoll (Lymphoprep., Nyegaard, Oslo, Norway) centrifugation. The oxidative burst response of these cells was measured for their ability to generate superoxide anion and was determined by a cytochrome c reduction assay. Preoperatively, all the plasma samples except one were free of endotoxin. The endotoxin levels reached 100 pg/ml 5 minutes after removal of the aortic crossclamp, and except in one sample they had decreased 24 hours after the operation. Studies on the generation of superoxide by neutrophils showed a decline in the response 5 minutes after removal of the aortic crossclamp and an enhancement of the response to f-Met-Leu-Phe by cells obtained from 11 of 12 patients 24 hours postoperatively. In vitro addition of bacterial lipopolysaccharide to blood from healthy individuals also enhanced the superoxide response of the neutrophils. We conclude that during cardiopulmonary bypass the circulating blood is contaminated by endotoxin and the neutrophils are primed for enhanced generation of oxygen radicals. The released oxygen radicals may be involved in the tissue damage observed in these patients. PMID- 2549308 TI - A cryogenic device for reversibly blocking transmission through small regions of the spinal cord white matter. AB - A simple cryogenic device is described. This device is capable of cooling neural tissue in contact with the probe and maintaining the tissue at the desired temperature for extended periods of time. The cold probe can thereby reversibly block neural transmission through small portions of the spinal cord white matter. Interruption of axonal transmission is achieved by placing the tip of the device in contact with the exposed surface of the spinal cord and cooling the tip of the probe to -1 to +2 degrees C. The investigator monitors the tip temperature and adjusts the pump rate to maintain a constant tip temperature. The cross-sectional area under the probe where effective transmission block is achieved is about 1.5 mm2 which approximates the size of a single funiculus in the cat thoracic spinal cord. The cryogenic device was constructed for less than $700. The properties of this device were studied in physiologic experiments in cats. This device reversibly, selectively and repeatedly blocked the ascending mass action potential in the dorsolateral funiculus, transmission through ascending spinal axons in the dorsal columns, transmission through axons of spinal dorsal horn cells, the descending inhibitory input to the dorsal horn and the activity of thalamic nociceptive neurons. The reversible cold block effects on single units were observed for the duration of the experiments (up to 18 h) with no detectable damage to the underlying tissue. The physiologic effects of the cold block were usually reversed a few minutes after rewarming, although in some cases it took up to 40 min for the complete reversal of the cold block. This cryogenic device is useful for studying spinal cord pathways. PMID- 2549309 TI - [Multiple glomangioma]. AB - A case of multiple glomus tumour is described in a 33 year old man. The clinical and histopathological features are discussed, with the review of the literature. We discuss the types of glomus tumour and its differential features. PMID- 2549310 TI - Glomus tympanicum tumors: contemporary concepts in conservation surgery. AB - The management of glomus tympanicum tumors is not a new subject. A review of the contemporary management concept is, however, needed for two reasons: 1. the recent literature has suggested radiation therapy as a treatment option in this surgical disease, and 2. patients who are referred to us for management of these tumors continue to exhibit errors in diagnosis and surgical approaches that are unnecessarily radical. The purpose of this paper is to review the diagnosis of these lesions and to prospectively differentiate them from the more extensive jugulare tumors. When type and extent of the tumor are properly determined, a surgical procedure can be planned that conserves as much normal ear anatomy and function as possible. Between May 1970 and July 1988, 60 patients with glomus tympanicum tumors were treated at the Otology Group, P.C., in Nashville, Tennessee. In 47 patients (78%), the tumors were removed using an extended facial recess approach. The external auditory canal wall was removed in seven patients (12%), and six patients underwent a transcanal approach. Total tumor removal was obtained in 90% of patients. Surgical management remains the mainstay of treatment for glomus tympanicum tumors. PMID- 2549311 TI - [Detection of IgG and IgM antibodies to herpes simplex virus type 1 and type 2 in human serum using an immunoenzyme method (ELISA) and the complement fixation reaction]. AB - On the basis of borderline optical density values (OD) in an enzyme-linked immunosorbent assay (ELISA) resulting from comparative testing of this method with complement fixation reactions (CF), determination was made of a set criteria for negative, weakly positive and positive results of IgG and IgM antibody detection of Herpes simplex type 1 (HSV 1), respectively type 2 (HSV 2). Of the 192 examined specimens by ELISA, 69.2% and 49.5% IgG HSV 1 and HSV 2 antibodies were found respectively. The number of sera containing IgM HSV 1 od HSV 2 antibodies was low (5.7%), since in the majority of cases the relapsing form of infection was involved. The sensitivity of ELISA to IgG in comparison with CF was slightly higher with the HSV 1 antigen (69.2:68.2%), and apparently lower with HSV 2 (49.5:28.2%). ELISA permits partial differentiation to be made between HSV 1 and HSV 2 antibodies due to a greater degree of antigen purity compared with the unpurified antigen used in CF. In fact, CF is known not to possess the sensitivity necessary to differentiate between HSV 1 and HSV 2 antibodies. For diagnostic purposes, ELISA findings could be more appropriately expressed quantitatively, in corresponding measurement units. PMID- 2549312 TI - Receptor autoradiographic localization of insulin-like growth factor-I (IGF-I) binding sites in human fetal and adult adrenal glands. AB - We report here the first evidence of insulin-like growth factor-I (IGF-I) binding sites in human fetal and adult adrenal glands, obtained at autopsy. Sections of tissue were incubated with 0.1 nM [125I]IGF-I and analyzed using [3H]Ultrofilm autoradiography with image analysis coupled to computerized microdensitometry. Specific binding sites of [125I]IGF-I were found to be localized in the definitive zone, fetal zone, and fetal medulla of the fetal adrenal glands. In the adult adrenal glands, the entire cortex and medulla were specifically labeled with [125I]IGF-I. Specific binding obtained at a concentration of 0.1 nM [125I]IGF-I to areas in the fetal and adult human adrenal glands was competitively displaced by unlabeled IGF-I, with an IC50 value of 0.34-2.54 nM, and 0.38-0.73 nM, respectively, whereas insulin was much less potent in displacing the binding. Acquisition of this knowledge will aid in studies on cell growth and steroid-catecholamines biosynthesis of the human adrenal gland. PMID- 2549313 TI - Desensitization of alpha-1 adrenergic receptor mediated smooth muscle contraction in aorta from endotoxic rats. AB - Desensitization of vascular smooth muscles in endotoxemia was studied using the aorta from intraperitoneally endotoxin-injected rats. The KCl- and phenylephrine induced contractions were significantly decreased in the endotoxic aorta compared to the control. In the endotoxic aorta the phenylephrine-induced contracture showed a gradual tension decrease after reaching a plateau and was attenuated by prior exposure to high concentration of phenylephrine, while KCl produced a sustained contraction and it was not affected by prior exposure to phenylephrine. The phenylephrine- and KCl-induced contractures of the control aorta showed stable plateaus and were not affected by prior exposure to phenylephrine. Neither diminished contractile force nor in vitro desensitization of phenylephrine contracture of isolated aorta was prevented by pretreatment of endotoxic rats with an alpha-adrenergic antagonist, phentolamine. These findings suggest that the contractile response to phenylephrine is easily desensitized in the endotoxic aorta compared to the control and neither this in vitro desensitization nor the diminution of contractile force is caused by in vivo exposure of aorta to a high concentration of catecholamines during endotoxemia. PMID- 2549314 TI - Methyl isobutyl amiloride: a new probe to assess the number of Na-H antiporters. AB - We measured the binding of [3H]-5-(N-methyl-N-isobutyl) amiloride (MIA) to purified rabbit renal brush border membranes. MIA binding was protein, temperature and time dependent with optimal binding at pH 8.0 or above. At low pH MIA binding was inhibited, suggesting competition between H+ ions and MIA for the MIA binding site. There was 70-80% specific binding which reached a plateau at 30 min and remained stable thereafter for 150 min. Scatchard analysis revealed one family of binding sites with Bmax of 3.4 +/- 0.4 pmoles/mg protein and Kd of 30.5 +/- 2.3 nM. MIA inhibited the Vmax of the Na-H antiporter (assessed by acridine orange quenching) in a dose dependent fashion with 100% inhibition at MIA concentration of 10(-3) M and this inhibition was greater than that of amiloride. We conclude that MIA, a potent inhibitor of the Na-H antiporter, displays a high percentage of specific binding to renal brush border membranes and can be used to assess the number of the Na-H antiporters. PMID- 2549315 TI - Effects of MSH on food intake, body weight and coat color of the yellow obese mouse. AB - Viable yellow obese mice (Avy/a) were treated for 10 days with 5, 15, 50 or 150 micrograms/d of either alpha-MSH or desacetyl-MSH. The half-maximal effect on weight gain occurred with a dose of 5 micrograms/d for desacetyl-MSH and at a 30 fold higher level of 150 micrograms/d for alpha-MSH. In contrast, the half maximal stimulation of eumelanin production by alpha-MSH occurred at 15 micrograms/d and with desacetyl-MSH at 150 micrograms/d, a 10-fold increase. Desacetyl-MSH produced a dose-related increase in the weight of muscle, as well as white and brown adipose tissue. Desacetyl-MSH and alpha-MSH both increased plasma corticosterone concentrations, but desacetyl-MSH was more potent. In a 2 x 2 factorial designed study, body weight was significantly increased in viable yellow mice only by treatment with desacetyl-MSH but in the lean animals, both alpha-MSH and desacetyl-MSH increased body weight. Food intake was significantly different between genotypes, and was stimulated by desacetyl-MSH. These studies demonstrate potent differences in biological actions on food intake, body weight, and a variety of organ weights between acetylated and desacetylated forms of MSH. PMID- 2549316 TI - Diazepam: endocrine effects and hypothalamic binding sites in the developing male and female rat. AB - The ontogeny of diazepam's endocrine effects in male and female rats, and of 3H diazepam binding in the hypothalami of both sexes was studied. Diazepam inhibited basal prolactin levels in 38 day-old male rats and, if prolactin levels were stimulated by Haloperidol the inhibition occurred in 28 day-old males, indicating that the hypoprolactinemic effect of the drug could be evidenced earlier if prolactin titers were high. The prolactin inhibition in females did not reach statistical significance at any studied age. Diazepam significantly released LH only in male rats at 12 days, showing thus, a period of special sensitivity of LH release to the drug. Benzodiazepine-hypothalamic binding sites increased in number from birth to puberty, reaching a plateau at 20 days of age. No sexual differences or changes in affinity were found throughout the studied period. These results suggest that the maturation of diazepam's hypoprolactinemic effect could be partially related to the increase in hypothalamic binding sites, whereas the sexual differences observed in diazepam's endocrine actions could be due to sexual differentiation of endocrine control mechanisms. PMID- 2549317 TI - Increased vascular contractile sensitivity to serotonin in spontaneously hypertensive rats is linked with increased turnover of phosphoinositide. AB - This study was conducted to determine if increased vascular contractile sensitivity to serotonin in spontaneously hypertensive (SHR) rats is linked with increased phosphoinositide turnover. Aortic and mesenteric artery rings from SHR exhibited 6.2- and 5.0-fold greater contractile sensitivity to serotonin than the aortic and mesenteric artery rings from normotensive Wistar-Kyoto (WKY) rats. Serotonin-induced turnover of phosphoinositide was measured by quantifying the accumulation of [3H] inositol labeled inositol monophosphate (IP), inositol bisphosphate (IP2) and inositol trisphosphate (IP3). Serotonin (3, 30, 200 microM) induced significantly greater accumulation of IP in SHR (279%, 590%, 895%) than in WKY (24%, 127%, 328%) aortic rings. Similarly, 3, 30 and 200 microM serotonin induced significantly greater accumulation of IP2 (118%, 241%, 451%) and IP3 (90%, 100%, 247%) in SHR than the accumulation of IP2 (15%, 58%, 122%) and IP3 (19%, 27%, 73%) in WKY aortic rings. Based on these data it is suggested that the greater vascular sensitivity to serotonin in SHR, at least in part, is attributable to increased turnover of phosphoinositide. PMID- 2549318 TI - Preventive effects of pyrroloquinoline quinone on formation of cataract and decline of lenticular and hepatic glutathione of developing chick embryo after glucocorticoid treatment. AB - When 0.25 mumol of hydrocortisone succinate sodium (HC) was administered to 15 day-old fertile eggs, almost all lenses of chick embryos treated with HC for 48 hr were classified as cataract stage IV-V (95%). A triple application of potassium pyrroloquinoline quinone (PQQ) (1.25 mumol/egg) at 3, 10 and 20 hr after HC treatment showed a preventive effect against the HC-induced cataract formation (I:45%, II:25%, III: 30%). PQQ also prevented the decline of GSH in the lens caused by HC. The decline of GSH in liver 24 hr after HC administration was prevented by PQQ. These data indicate that PQQ can modify HC-induced effects and that the preventive effect of PQQ against HC-induced decline of hepatic GSH seemed to influence HC-induced events in lens. PMID- 2549319 TI - Calcium-mobilizing receptors, polyphosphoinositides, generation of second messengers and contraction in the mammalian iris smooth muscle: historical perspectives and current status. AB - It is well established now that activation of Ca2+ -mobilizing receptors results in the phosphodiesteratic breakdown of phosphatidylinositol 4,5-bisphosphate (PIP2), instead of phosphatidylinositol (PI), into myoinositol 1,4,5 trisphosphate (IP3) and 1,2-diacylglycerol (DG). There is also accumulating experimental evidence which indicates that IP3 and DG may function as second messengers, the former to mobilize Ca2+ from intracellular sites and the latter to activate protein kinase C (PKC). In this review, I have recounted our early studies, which began in 1975 with the original observation that activation of muscarinic cholinergic and adrenergic receptors in the rabbit iris smooth muscle leads to the breakdown of PIP2, instead of PI, and culminated in 1979 in the discovery that the stimulated hydrolysis of PIP2 results in the release of IP3 and DG and that this PIP2 breakdown is involved in the mechanism of smooth muscle contraction. In addition, I have summarized more recent work on the effects of carbachol, norepinephrine, substance P, the platelet-activating factor, prostaglandins, and isoproterenol on PIP2 hydrolysis, IP3 accumulation, DG formation, myosin light chain (MLC) phosphorylation, cyclic AMP production, arachidonic acid release (AA) and muscle contraction in the iris sphincter muscle. These studies suggest: (a) that the IP3-Ca2+ signalling system, through the Ca2+ -dependent MLC phosphorylation pathway, is probably the primary determinant of the phasic component of the contractile response; (b) that the DG PKC pathway may not be directly involved in the tonic component of muscle contraction, but may play a role in the regulation of IP3 generation; (c) that there are biochemical and functional interactions between the IP3-Ca2+ and the cAMP second messenger systems, cAMP may act as regulator of muscle responses to agonists that exert their action through the IP3-Ca2+ system; and (d) that enhanced PIP2 turnover is involved in desensitization and sensitization of alpha 1-adrenergic- and muscarinic cholinergic-mediated contractions of the dilator and sphincter muscles of the iris, respectively. The contractile response is a typical Ca2+ -dependent process, which makes smooth muscle an ideal tissue to investigate the second messenger functions of IP3 and DG and their interactions with the cAMP system. PMID- 2549320 TI - Receptors for atrial natriuretic peptide (ANP) and regulation of thyroglobulin secretion by ANP in human thyroid cells. AB - Specific binding sites for atrial natriuretic peptide (ANP) were identified and characterized in primary cultures of human thyroid cells. Saturation analysis using [125I] alpha rat ANP as the ligand showed a single class of high affinity binding (Kd = 0.2 nM) which was inhibited by atriopeptin I and the alpha -human form of ANP, but not by a C-terminal fragment of the peptide. The number of ANP binding sites in these cultures was not altered by the thyroid hormone concentration of the medium. In a dose-response experiment, thyro-globulin secretion was significantly reduced in the presence of 0.01 nM ANP and was maximally reduced (to 25% of control value) with 10 nM ANP. Cyclic GMP production was increased threefold in the presence of 100 nM ANP, but was unchanged with lower doses (0.01 and 0.1 nM) of the peptide. The finding of receptors in thyroid follicular cells suggests a hitherto unrecognized role of ANP in the thyroid gland. PMID- 2549321 TI - Prenatal ethanol exposure reduces the effects of excitatory amino acids in the rat hippocampus. AB - Chronic alcohol ingestion during pregnancy can lead to the Fetal Alcohol Syndrome (FAS), a disorder marked by learning disabilities. A rat model of FAS was used by introducing pregnant Sprague-Dawley rats to a liquid diet containing 35% ethanol derived calories (E), while a second group was pair-fed an isocaloric liquid diet without ethanol (P). A third group of pregnant dams received ad libitum lab chow (C). At parturition, pups from the E and P groups were cross-fostered by C mothers and all groups received lab chow. During adulthood, male offspring were sacrificed and hippocampal and prefrontal cortical slices were prelabeled with [3H] inositol. Phosphoinositide (PI) hydrolysis was determined by measuring the accumulation of [3H]inositol phosphates in the presence of LiCl in response to activation of various excitatory amino acid (EAA) receptors. In hippocampal slices, ibotenate- and quisqualate-induced PI hydrolysis was reduced in E compared to P and C animals. Moreover, the inhibitory effect of N-methyl-D aspartate (NMDA) on carbachol-induced PI hydrolysis, evident in P and C animals, was completely abolished in the hippocampus of E animals. In contrast, in the prefrontal cerebral cortex, this inhibitory effect of NMDA prevailed even in the E animals. The evidence suggest that prenatal ethanol exposure alters the activity of EAA receptors in the hippocampal generation of 2nd messengers. PMID- 2549322 TI - Ac-Tyr1hGRF discriminates between VIP receptors from rat liver and intestinal epithelium. AB - The vasoactive intestinal peptide (VIP) stimulates adenylate cyclase activity in rat liver and intestinal epithelium with low and high efficacy, respectively. The human growth hormone releasing factor (hGRF) derivative with acetylated N terminus e.g. Ac-Tyr1hGRF binds to VIP receptors in both tissues with a similar affinity. However, Ac-Tyr1hGRF is a partial VIP agonist with high intrinsic activity in liver (50% that of VIP) whereas it behaves as a VIP antagonist in intestine. These results further argue for a possible heterogeneity of VIP receptor-coupled adenylate cyclase among tissues on a pharmacological basis. PMID- 2549324 TI - Prolonged IgM antibodies and histopathological evidence of chronicity in hepatitis A. AB - The case of a young man with hepatitis A and a chronic course is presented. The patient received a short course of steroid therapy for recurrence of symptoms following acute hepatitis A. Thereafter, liver enzymes have remained marginally elevated for 4 years and annual liver biopsies have shown evidence of chronicity. HAV IgM Ab persisted for 1034 days with subsequent development of HAV IgG Ab. The possibility of other viruses in the aetiology and the role of steroids in the development of chronicity are discussed. PMID- 2549323 TI - Influence of dietary fat composition on intestinal absorption in the rat. AB - Omega-3 fatty acids influence the function of the intestinal brush border membrane. For example, the omega-3 fatty acid eicosapentaenoic acid (20:5 omega 3) has an antiabsorptive effect on jejunal uptake of glucose. This study was undertaken to determine whether the effect of feeding alpha-linolenic acid (18:3 omega 3) or EPA plus docosahexaenoic acid (22:6 omega 3) on intestinal absorption of nutrients was influenced by the major source of dietary lipid, hydrogenated beef tallow or safflower oil. The in vitro intestinal uptake of glucose, fatty acids and cholesterol was examined in rats fed isocaloric diets for 2 weeks: beef tallow, beef tallow + linolenic acid, beef tallow + eicosapentaenoic acid/docosahexaenoic acid, safflower oil, safflower oil + linolenic acid, or safflower oil + eicosapentaenoic acid/docosahexaenoic acid. Eicosapentaenoic acid/docosahexaenoic acid reduced jejunal uptake of 10 and 20 mM glucose only when fed with beef tallow, and not when fed with safflower oil. Linolenic acid had no effect on glucose uptake, regardless of whether it was fed with beef tallow or safflower oil. The jejunal uptake a long-chain fatty acids (18:0, 18:2 omega 6, 18:3 omega 3, 20:4 omega 6, 20:5 omega 3 and 22:6 omega 3) and cholesterol was lower in safflower oil than with beef tallow. When eicosapentaenoic acid/docosahexaenoic acid was given with beef tallow (but not with safflower oil), there was lower uptake of 18:0, 20:5 omega 3 and cholesterol. The demonstration of the inhibitory effect of linolenic acid or eicosapentaenoic acid/docosahexaenoic acid on cholesterol uptake required the feeding of a saturated fatty acid diet (beef tallow).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549326 TI - [Local radiometry of tumors of the breast]. AB - The authors propose a radionuclide method of diagnosis of breast tumors based on focal radiometry of 99mTc-pertechnetate accumulated in them after injection. Radiometry is done with a scanner with a pulse counter. The ratio of pulse counts accumulated in a tumor, to this indicator on the contralateral side makes it possible to determine a tumor type. This method is of diagnostic importance for tumors with a diameter less than 2 cm, their individual sections and also for lymph nodes. PMID- 2549325 TI - [Regional and general contractile function of the myocardium following myocardial infarct]. AB - A total of 93 patients with myocardial infarction were investigated 3-4 mos. after the beginning of disease by the method of radionuclide ventriculography. Total and regional ejection fractions (for 7 zones), left ventricular functional volumes (end-systolic, end-diastolic and stroke volumes), rates of contraction and expulsion were determined. A certain level of left ventricular pump function in patients after myocardial infarction was determined by the ratio of the number of hypokinetic segments and the expression of a decrease in their contractility and normo- and hyperkinetic segments which are the main compensatory factor. Variations in the structure of formation of the total ejection fraction with relation to a site of myocardial infarction were revealed. Compensatory significance of hyperkinetic zones is decreased with a growing degree of expression of contraction asynergia. PMID- 2549327 TI - [Comprehensive diagnosis of cancer of the ovaries]. AB - Combined examination of 96 patients with different gynecological diseases included methods of ultrasound tomography, scintigraphy of the small pelvis with 99mTc-pyrophosphate, immunoscintigraphy and radioimmunoassay for determination of the blood level of CA 125. New diagnostic methods for ovarian cancer using 131I labeled monoclonal antibodies and an algorithm for data processing were developed. Of 96 patients 39 had ovarian cancer, 6 patients--uterine tumors, and 51 patients (controls) with nontumorous and benign lesions of the ovaries and uterus. The diagnostic effectiveness of these methods was determined by the indices of sensitivity, specificity and accuracy. The radioimmunoassay for determination of CA 125 possessed the highest sensitivity, ultrasound tomography showed the highest specificity and accuracy. Immunoscintigraphy with Imacis-2 was shown to possess the highest diagnostic effectiveness and unique informative value. An optimum algorithm for the use of the above methods in patients with suspected ovarian cancer was proposed on the basis of these investigations. PMID- 2549328 TI - Involvement of voltage-operated calcium channels in alpha-melanocyte-stimulating hormone (alpha-MSH) release from perifused rat hypothalamic slices. AB - The contribution of voltage-operated calcium (VOC) channels in the mechanism of release of alpha-melanocyte-stimulating hormone (alpha-MSH) from hypothalamic neurons was investigated using perifused rat hypothalamic slices. The stimulatory effect of potassium (50 mM) on alpha-MSH release was completely blocked by cadmium (1 mM) a calcium competitor which indifferently blocks T-, L-and N-type VOC channels. To determine the nature of calcium conductances involved in K+ evoked alpha-MSH release, we have investigated the effect of a VOC channel agonist and 3 antagonists on the secretion of the neuropeptide. Administration of synthetic omega-conotoxin fraction GVIA (1 microM), a peptide toxin which blocks both N- and L-type VOC channels, reduced by 33% K+-induced alpha-MSH release. In contrast, the 1,4-dihydropyridine (DHP) antagonist nifedipine, at concentrations up to 100 microM, did not affect the response of hypothalamic alpha-MSH neurons to depolarizing concentrations of KCl. In addition, the secretion of alpha-MSH induced by high K+ concentrations was not reduced by nifedipine (10 microM) in the presence of diltiazem (1 microM), a benzothiazepine derivative which increases the affinity of the DHP antagonist for L-type VOC channels. The DHP agonist BAY K 8644 (0.1-10 microM) did not modify the early phase of the response of alpha-MSH neurons to K+-induced depolarization. In contrast BAY K 8644 (1 or 10 microM) significantly prolonged the duration of K+-induced alpha-MSH release. This sustained release of alpha-MSH induced by BAY K 8644 (10 microM) was totally suppressed by nifedipine (10 microM).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549329 TI - Dual virus etiology of age-dependent poliomyelitis of mice. A potential model for human motor neuron diseases. PMID- 2549330 TI - Enhancement of peripheral nerve regeneration by pharmacological activation of the cyclic AMP second messenger system. AB - This paper reviews the history of attempts to study peripheral nerve regeneration, focusing upon pharmacologic agents that may prove to be useful clinically. In particular, forskolin is described as a model for such an agent, and its mechanism of action, as a stimulator of the cyclic AMP second messenger system, is described. PMID- 2549331 TI - Oxygen consumption and guanosine diphosphate binding by fetal brown adipose tissue in diabetic pregnancy. AB - Oxygen consumption and 3H-guanosine diphosphate (GDP) binding were determined in brown adipocytes and mitochondria from 28-day gestation fetuses of alloxan diabetic rabbit does and saline-injected controls. Maternal diabetes was classified as severe or mild determined by whether maternal blood glucose values were greater or less than 200 mg/dL, respectively, at death. Basal oxygen consumption and adipocyte diameters did not vary among groups. A significant reduction in maximal norepinephrine (NE) stimulated O2 consumption by fetal brown adipose tissue (BAT) cells was seen in offspring of severely diabetic pregnancies when compared with control values (248 +/- 53 +/- v482 +/- 32 microL O2/10(6) cells/h; P less than .005). In contrast, a significant increase in maximal NE stimulated O2 consumption by fetal BAT cells occurred in offspring of mild diabetic pregnancies (807 +/- 60, P less than .001 v controls). A highly significant inverse correlation between serum glucose levels and maximal O2 consumption by fetal BAT was observed in fetuses from mild and severe diabetic pregnancies (r = -.98, P less than .005), and there was no correlation between these two parameters in offspring of normal pregnancies. A significant inverse correlation was observed between maximal O2 consumption by fetal BAT cells and serum insulin levels in offspring of both control and diabetic pregnancy (r = .74; P less than .02). Tissue cytochrome oxidase activity was lower in offspring of severely affected diabetic does, indicating a reduction in BAT mitochondrial content compared with controls. BAT mitochondria from fetuses of severely diabetic does exhibited reduced 3H-GDP capacity, which was 2.5-fold lower than controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549332 TI - A proposed mechanism of the mitogenic action of fluoride on bone cells: inhibition of the activity of an osteoblastic acid phosphatase. AB - Fluoride (F) is a potent inhibitor of osteoblastic acid phosphatase activity with an apparent Ki value (10 to 100 mumol/L) that corresponds to F concentrations that increase bone cell proliferation and bone formation in vivo and in vitro. This high sensitivity of acid phosphatase to F inhibition appeared to be specific for skeletal tissues. Mitogenic concentrations of F did not increase cellular cAMP levels but significantly stimulated net protein phosphorylation in intact calvarial cells and in isolated calvarial membranes. These concentrations of F also stimulated net membrane-mediated phosphorylation of angiotensin II (which contains tyrosyl but no seryl or threonyl residues), suggesting that some of the F-stimulated protein phosphorylations could occur on tyrosyl residues. F had no apparent effect on thiophosphorylation of membrane proteins, suggesting that the F-stimulated net protein phosphorylation in bone cells was probably not mediated via activation of protein kinases. Orthovanadate or molybdate at concentrations that inhibit bone acid phosphatase activity also stimulated bone cell proliferation, supporting the idea that inhibition of bone acid phosphatase would lead to stimulation of bone cell proliferation. Mitogenic concentrations of F potentiated the mitogenic activities of insulin, EGF, and IGF-1 (ie, growth factors the receptors of which are tyrosyl kinases) to a greater extent than they potentiated the action of basic FGF (a growth factor that does not appear to stimulate tyrosyl protein phosphorylation). Based on these findings, a model is proposed for the biochemical mechanism of the osteogenic action of F in which F stimulates bone cell proliferation by a direct inhibition of an osteoblastic acid phosphatase/phosphotyrosyl protein phosphatase activity, which in turn increases overall cellular tyrosyl phosphorylation, resulting in a subsequent stimulation of bone cell proliferation. PMID- 2549334 TI - Preparation of simian virus 40 minichromosomes. PMID- 2549335 TI - Electron microscopy of simian virus 40 minichromosomes. PMID- 2549333 TI - Evidence for the existence of a GTP-dependent factor in hepatic cytosol that stimulates cyclic AMP binding: possible role in the modulation of cyclic AMP action. AB - GTP, in physiologic concentration (10(-4) mol/L), enhances cAMP binding to an Mr 57,000 binding protein (BP) in hepatic cytosol, which probably is the phosphorylated receptor subunit of protein kinase II (PK II). When we attempted to separate PK II from other hepatic cytosol proteins by DEAE-cellulose chromatography, we observed that GTP caused little stimulation of [3H]cAMP binding in an eluate fraction (110 to 170 mmol/L KCI), which was rich in PK II but did stimulate cAMP binding to the 210 to 325 mmol/L KCI fraction, which also contains PK II. This suggested that the latter fraction might contain a cofactor necessary for GTP stimulation of cAMP binding, which was lacking in the 110 to 170 mmol/L KCI fraction. Cyclic AMP BP in the 210 to 325 mmol/L fraction was removed by absorption onto cAMP agarose in the presence of 325 mmol/L KCI. When an aliquot of the BP-poor fraction, containing the putative cofactor, was added to the 110 to 170 mmol/L fraction containing PK II, the addition of 10(-4) mol/L GTP to the mixture increased [3H]cAMP binding by more than 80%. Cofactor activity could be extracted from the 210 to 325 mmol/L eluate by adsorption onto cAMP agarose in the presence of 10 mmol/L K phosphate, and eluted with 100 mmol/L KCI, suggesting that the cofactor may bind to the cAMP BP under appropriate circumstances. Addition of this eluted cofactor fraction to the 110 to 170 mmol/L fraction in the presence of 10(-4) mol/L GTP, increased the specific binding of [3H] cAMP more than twofold. Pretreatment of the cofactor fraction with trypsin eliminated this effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549336 TI - Application of nucleases to visualizing chromatin organization at replication forks. PMID- 2549337 TI - Analysis of nucleosome positioning by indirect end-labeling and molecular cloning. PMID- 2549338 TI - Affinity isolation of replicating simian virus 40 chromosomes. PMID- 2549339 TI - Isolation and characterization of histones. PMID- 2549340 TI - Purification of Xenopus egg nucleoplasmin and its use in chromatin assembly in vitro. PMID- 2549341 TI - The occurrence of nicotinamide adenine dinucleotide glycohydrolase and the serum opacity reaction in groups B, C, F and G streptococci. AB - The frequency of the serum opacity reaction (SOR) and nicotinamide adenine dinucleotide glycohydrolase (NADase) production in non-A groups of beta haemolytic streptococci isolated from humans and animals was surveyed. The SOR was positive with five of eighteen group B isolates (28%), four of fifteen group C (27%), two of five group F (40%), and thirty-seven of sixty-eight group G (54%) isolates. NADase activity, in addition to SOR, was found in three of eighteen group B isolates (17%), three of fifteen group C (20%), forty of sixty-eight group G (59%), and none of the group F isolates. The SOR was produced by forty eight (45%) and NADase was produced by fifty-one (48%) of the one hundred and six isolates of non-group A beta-haemolytic streptococci. Twenty-five percent of the isolates were both NADase and SOR positive. PMID- 2549342 TI - Phosphomonoesterase activity of the cyanobacterium (blue-green alga) Calothrix parietina. AB - Cellular and extracellular phosphomonoesterase activities were compared in Calothrix parietina D550, a strain whose original environment has been studied in detail. Activity in both fractions became detectable at about the same stage in batch culture. Differences in the influence of environmental factors between the two were slight, suggesting a common origin. The optimum temperatures for cellular and extracellular activities were 40 degrees C and 30 degrees C, respectively, and the upper limits for detectable activity were 80 degrees C and 65 degrees C. The pH optimum for both cellular and extracellular activity was 10.0-10.2. When P-limited cultures were tested with p-nitrophenyl phosphate (pNPP) as substrate, Km values for cellular and extracellular activities were 43 and 33 microM pNPP, respectively. Eleven ions were tested for their influence on activity. In most cases the effect was low or negligible at concentrations likely to be present in nature or freshwater laboratory media. Where obvious effects occurred, these were usually apparent at lower concentrations with extracellular than cellular activity. One mM Ca led to a 40% increase in extracellular activity in comparison with 0.1 mM Ca, but had no effect on cellular activity. However, inorganic phosphate, which had a marked inhibitory effect at concentrations above 10 microM, brought about a similar response with cellular and extracellular activities (approximately 60% decrease with 100 microM). PMID- 2549343 TI - Transcription mapping of glycoprotein I (gpI) and gpIV of varicella-zoster virus and immunological analysis of the gpI produced in cells infected with the recombinant vaccinia virus. AB - In order to determine the transcripts of gpI and gpIV of varicella-zoster virus (VZV), RNA was isolated from human embryonic fibroblast cells infected with VZV and Northern blot analysis was carried out using cloned DNA probes of unique short region including gpI and gpIV genes. The analysis of RNA revealed two discrete transcripts of 3.6 and 2.15 kilobases (kb) and three transcripts of 3.6, 2.9, and 1.6 kb which hybridized to DNA probes covering the gpI and gpIV region, respectively. Next, mRNAs were hybrid-selected, translated in vitro and the polypeptide products were immunoprecipitated with antibodies against these glycoproteins. The polypeptides with a molecular weight of 70,000 (70K) and 37K which were in vitro translational products of mRNA hybrid-selected with the DNA clone covering gpI and gpIV were detected using antibodies against gpI and gpIV, respectively. The result showed that the 70K polypeptide is presumably the translational product of 2.15 kb mRNA and the 37K polypeptide is that of 1.6 kb mRNA. DNA fragment encoding gpI or gpIV was inserted into vaccinia virus DNA and the recombinant viruses, mO74 (gpI) and mO39 (gpIV), were used for immunological analysis. In consequence, the gpI derived from cells infected with mO74 showed antigenic characteristics similar to those of gpI from VZV-infected cells as determined from the immunoprecipitation pattern, although the molecular weight of each polypeptide was different, and antibody produced in rabbits infected with recombinant virus had a high neutralizing activity, when the reaction was performed with complement. This suggested that gpI plays an important role for protection and recovery from VZV infection. PMID- 2549344 TI - Role of pertussigen (pertussis toxin) on the mouse protective activity of vaccines made from Bordetella species. AB - Pertussigen [pertussis toxin (Ptx)] from Bordetella pertussis, when detoxified, induces protection in mice to intracerebral challenge (ic) with virulent B. pertussis. In its native form, minute nonprotective doses promote the development of immunity induced by other antigens of B. pertussis. As little as 4 ng of Ptx, given with a nonprotective dose of 8 X 10(7) killed cells of the phase III Sakairi strain, promoted detectable protection to ic challenge. Native Ptx in doses of 0.4 to 400 ng did not protect mice, and vaccines made from strains not producing Ptx induced only weak protection. The marked enhancing action of Ptx was also observed with 5 micrograms of purified filamentous hemagglutinin and with vaccines made from other species of the Bordetella genus, such as B. parapertussis and B. bronchiseptica, but it was not observed with B. pertussis endotoxin. In addition, Ptx was still effective when given as late as 7 days after the vaccine. Antibodies to surface antigens of the challenge strain were demonstrated in sera of mice immunized with vaccines prepared with the different Bordetella species tested, but antibodies to Ptx were detected only in the sera of mice immunized with the wild-type B. pertussis strains. Glutaraldehyde detoxified Ptx does not have this action. Pretreatment of normal mice with Ptx, also enhanced the protective action of a mouse antiserum to a wild-type strain of B. pertussis. These observations show that antigens other than Ptx are responsible for the protection, and that Ptx acts non-specifically to enhance the mouse protective action of those antigens. PMID- 2549345 TI - Human parvovirus (HPV/B19) infection with purpura. AB - A 33-year-old man complained of purpura (petechial hemorrhage) in chelidons, poples, axillae, and bilateral chest in addition to other symptoms such as lumbago, arthralgia, muscular pain, and fever. On the next day of the onset, human parvovirus (HPV/B19) antigen and HPV/B19 DNA were detected in his serum, and twelve days later IgM antibody to HPV/B19 became detectable. This case supports the relationship between purpura and HPV/B19 infection. PMID- 2549346 TI - Characterization of a nalidixic-acid-resistant mutant of Escherichia coli as a strict aerobe. AB - An Escherichia coli mutant, C18, which plates at an efficiency of 5.0 x 10(-4) under anaerobic condition, was isolated among spontaneous nalidixic-acid resistant mutants. This strict aerobic mutation was mapped by P1 cotransduction with a gyrA linked transposon Tn10 and found to be at the gyrA gene. A low degree of superhelicity of pBR322 DNA isolated from C18 was demonstrated by agarose gel electrophoresis with various concentrations of ethidium bromide. The superhelical density of pBR322 isolated from C18 was 80% of the value of pBR322 isolated from wild-type bacteria cultured under aerobic condition, and 50% cultured under anaerobic condition. These results lead us to conclude that a certain mutation of the gyrA gene causes a decrease in DNA superhelicity and prevents anaerobic growth. PMID- 2549348 TI - Prevalence of antibody to human herpesvirus 6 in women and children. AB - The antibody prevalence to human herpesvirus 6 (HHV-6) was compared between pregnant women and control women of similar ages in Thailand. No significant difference was detected in the antibody positive rate and antibody titers between both groups. The antibody titers in sera collected from pregnant women at 1st and 3rd trimester remained unchanged. Next, the antibody prevalence in infants were examined and the positive rate decreased until 3 months and started to increase from 6 months after birth. The present results suggest that the reactivation of HHV-6 might not occur during pregnancy and this virus infects infants postnatally. PMID- 2549347 TI - Protection of OK-432, a Streptococcus pyogenes preparation, against lethal infection of mice with herpes simplex virus. AB - We have studied the protective effect of OK-432, a biological response modifier (BRM) of Streptococcus pyogenes origin, on the lethal infection of mice with herpes simplex virus (HSV)-1. A single intraperitoneal (i.p.) injection of more than 10 micrograms of OK-432, when given at least two days before the infection, gave a marked effect yielding nearly 100% protection against ordinarily lethal infection. The protection was independent of the amount of infected virus inoculated. When given after the infection, the agent even at the maximal dose (100 micrograms), produced only a marginal effect. A single i.p. administration of OK-432 augmented the natural killer (NK) activity of peritoneal exudate cells and spleen mononuclear cells in mice 2 to 3 days after injection of OK-432, coinciding with the times when it induced a survival effect on HSV-infection. Treating OK-432-treated mice with a combination of an anti-macrophage agent, silica, and an anti-NK cell agent, anti-asialo GM1 serum, before infection diminished the antiviral effect of OK-432. The OK-432 protection against HSV infection was also markedly diminished in athymic nude mice. Thus, the protective effect of OK-432 on lethal HSV infection seems to be based on the activation of NK cells, macrophages, and T lymphocytes. PMID- 2549350 TI - Epidemiology of hepatocellular carcinoma in French Polynesia. PMID- 2549349 TI - Specific immunoglobulin response in mice immunized with porins and challenged with Salmonella typhi. AB - Specific antiporin antibody (IgG, IgM, and IgA) response was studied in control, infected, immunized-infected, and immunized mice. The activity of specific IgG immunoglobulins was found to be the highest in immunized and immunized-infected groups in which 87.5% protection had been observed by laboratory potency test in mice; the next-highest activities were those of IgM and IgA immunoglobulins. However, in the infected group the activity of specific IgM and IgG immunoglobulins was almost similar. PMID- 2549352 TI - Ganciclovir. PMID- 2549351 TI - [Peripheral neuropathy caused by incongruous posture in shoe manufacture workers]. AB - Nine cases of local neuropathy are described in shoe-manufacture workers. Clinical and electrophysiological examination excluded diseases such as toxic polyneuropathies due to n-hexane. Specific lesions of the ulnar nerve were located near the elbow where the forced and uncomfortable posture facilitated the compression of the nerve in the cubital tunnel. The severity of the neuropathies ranged from relatively slight, with paresthesia of some of the fingers, to evident hypotrophy of the inter-osseous muscles of the hand. All the 9 cases sewed shoe uppers using sewing machines operating 25 cm above the arm support, thus obliging the operator to work with one or both elbows resting on the support, with the arms and shoe-upper held up. Five cases presented a neuropathy of the right arm, 3 of the left arm, and in one case the lesions were bilateral. The features of the work station are described and the possible promoting factors and the ergonomic pathogenesis of these neuropathies are discussed. PMID- 2549353 TI - Variable versus fixed modulation of proton beams for treatments in the cranium. AB - Dose distributions in the cranium with fixed and with variably modulated proton beams were compared. The variable modulation was designed to tailor the proximal high-dose region of each field to the target volume surface whereas the fixed modulation beams had a constant modulation determined by the greatest extent of the target. Dose-volume histograms of normal tissues were compared, as were the estimated complication probabilities. Twelve patients with chordomas or chondrosarcomas of the base of skull who had been treated to approximately 70 cobalt Gray equivalent (CGE) were evaluated. Dose distributions of the actual treatments were compared to those which would have been delivered had the proton beams been variably modulated; two patients for whom x-ray plans were available were also evaluated. The greatest difference in dose between the variable and fixed modulation proton beams, averaged over all the patients, was 13.8 CGE (8.0 18.0 CGE range). Much of this reduction occurred in the brain, particularly the temporal lobes. In those temporal lobes receiving significant doses, variable modulation reduced the volume receiving more than 54 CGE by 3.0 cc; all temporal lobes had at least a 5 CGE difference to some portion, half had more than 10 CGE and three more than 15 CGE difference to some portion. The optic structures, brainstem and spinal cord received from 1 to 3 CGE less dose with the variability modulated beams. Eight of the parotid glands received more than 20 CGE to more than half their volume with the fixed modulation beams; in these, variable modulation reduced the mean dose by 5.3 CGE. The reduction in integral dose with variable as compared to fixed modulation was in the range 3 to 12%; this gain was considerably less than the gain for uniformly modulated proton beams over x-rays in the two patients for whom x-ray plans were available. PMID- 2549354 TI - Temperature measurements during ultrasound hyperthermia. AB - In this study the ultrasonic field distortions and the temperature measurement errors caused by temperature sensing probes were investigated. It was found that probes with diameters equal to or greater than 1/2 of the square root of the wavelength (lambda) scatter and reflect the waves, and thus distort the field significantly. Smaller probes down to lambda 1/2/5 in diameter had a detectable effect which was very local and, therefore, will probably not have a significant effect on the overall temperature distribution. When the temperature measurement errors were studied, even the smallest probes showed some self-heating artifact and its magnitude depended on the probe size, material, structure, orientation, and the operating frequency. This error was small with most of the nonultrasound absorbing probes (such as bare wires or stainless steel needle probes), but significant with plastic shielded thermocouples. The energy absorption associated with plastic coated probes can be reduced by orienting the probe parallel to the beam, by scanning the focus in such a manner that the focus is not passing on the sensor, and by inserting the probe in a stainless steel catheter. The temperature measurement error can also be reduced by covering the plastic probe with a high- (or low-) acoustical impedance material around the sensor. This would scatter the sound around the probe, thus preventing energy absorption in the plastic. This arrangement can preserve the desirable properties of plastic probes (mainly flexibility, strength, and electrical isolation from the patient) while at the same time allowing fairly absorption artifact free measurements. PMID- 2549355 TI - Use of a fiber-containing enteral formulation in an AIDS patient. AB - The value of aggressive nutritional support in the patient with AIDS requires further study. As medical treatment modalities become more sophisticated, the etiology of complications that are associated with AIDS are more clearly understood and can be more effectively treated. Survival rates of greater than one and one-half years after an episode of pneumocystis are now more common. As clinicians become more familiar with AIDS and as new medical treatments are discovered, there is reason to believe that a gradual improvement in survival and quality of life will continue. Therefore, further research is needed to determine the point at which nutritional therapy is truly beneficial in the AIDS population. PMID- 2549356 TI - RTCs restructured with compromises. PMID- 2549357 TI - Questions and answers on liability insurance. PMID- 2549358 TI - Sexual harassment on the job. PMID- 2549360 TI - Neurologic manifestations of gastrointestinal disease. AB - The neurologic manifestations of gastrointestinal disease are generally thought to be uncommon, although an increasing number of previously unidentified associations are being established. These neurologic disorders may result from nutritional or non-nutritional causes. In the absence of clear malnutrition, it is likely that many of these disorders are underdiagnosed. As an example, Wernicke's encephalopathy is found at autopsy in as many as 2 per cent of brains, a very high percentage, given the rare recognition during life. The likely underdiagnosis of nutritional neurologic disorders is unfortunate because many are treatable and, more importantly, are preventable if malabsorption is suspected and appropriate supplementation initiated. For the neurologist, familiarity with the occasional association between neurologic abnormalities and specific gastrointestinal disorders is important, as is familiarity with the neurologic characteristics of disorders, such as Whipple's disease, that may present as isolated neurologic syndromes without gastrointestinal symptoms or signs. Renewed interest in selective deficiency states has resulted in identification of causative factors in several neurologic syndromes of previously presumed degenerative etiology. Recognition of the potential neurologic consequences of prolonged deficiency states also is important for the internist, because many of the syndromes are poorly reversible once symptomatic. The benefits of prevention invariably exceed those of treatment. PMID- 2549359 TI - [Renal function and sleep apnea syndromes]. AB - Obstructive sleep apnea (OSA) patients have increased diuresis and natriuresis during sleep. In order to investigate the possible mechanisms of these changes in renal function, 35 consecutively diagnosed OSA patients were studied during sleep before and during nasal continuous positive airway pressure (CPAP) treatment, and were compared with 23 non-snoring controls. The excretion of urine and of electrolytes was increased before treatment and normalized with nasal CPAP treatment. The mechanism involved seems to be decreased sodium reabsorption at the level of the ascending limb of the loop of Henle. The observed increase in cyclic guanosine monophosphate excretion supports the hypothesis of increased atrial natriuretic peptide release during sleep in OSA patients. PMID- 2549361 TI - The remote effects of cancer on the nervous system. AB - Paraneoplastic syndromes are heterogeneous in their clinical presentations and their associations with particular tumor types and are an important part of the differential diagnosis of neurologic dysfunction in patients with or without a known neoplasm. Patients presenting with one of the more distinctive syndromes, such as subacute cerebellar degeneration, opsoclonus-myoclonus, and the Lambert Eaton syndrome, should undergo a careful evaluation for the presence of an occult malignancy. The importance of looking for a monoclonal gammopathy in patients with certain polyneuropathies and motor neuron syndromes is also becoming clear. At this time, an autoimmune pathogenesis has been clearly demonstrated only for the Lambert-Eaton syndrome. Specific autoantibodies in other syndromes appear to be valuable diagnostic markers for the presence of an underlying malignancy, but the actual role of these antibodies in producing tissue damage and clinical disease is still unknown. PMID- 2549362 TI - Neurologic complications of renal disease. AB - Renal failure and its treatment are associated with a number of neurologic complications that must be differentiated from the nervous system complications of the disease leading to renal failure. Uremic encephalopathy is characterized by clinical signs of depressed brain function coexisting with excitation, often in the form of generalized epileptic seizures. Peripheral neuropathy, due to axonal involvement, is common and is characterized by ascending sensory and motor dysfunction. The treatment of renal failure also may lead to the development of neurologic abnormalities in the form of dialysis disequilibrium characterized by headache, nausea, irritability that may progress to seizures, coma, and death, which is caused by the entry of free water into the brain and swelling. Dialysis dementia, caused by the toxic effects of aluminum, is now rare. Renal transplant recipients may develop cerebrovascular disease, infections by opportunistic organisms, or malignant neoplasms, particularly primary lymphoma of the brain. As transplant recipients live longer and more operations are performed, additional complications may be seen in the future. PMID- 2549363 TI - [Protective effects of DBcAMP on acute liver failure induced by D-galactosamine in rats]. AB - We studied protective effects of Dibutyryl cyclic AMP (DBcAMP) which is a permeable form of cyclic AMP (cAMP), the intracellular second messenger, on D galactosamine (D-Gal, 1.5g/kg i.p.) induced acute liver failure. Experimental animals were divided into four groups: Group I; DBcAMP was administered before (7.5mg/kg) and after (7.5mg/kg) D-Gal treatment, group II; only before (15mg/kg) D-Gal treatment, group III; only after (15mg/kg) D-Gal treatment, and group IV; no DBcAMP was administered. Effects of DBcAMP were evaluated according to survival rates, serum biochemical findings and hepaplastin test (HPT), ATP levels in liver tissues, hepatic blood flow, and histological findings. The survival rates after D-Gal treatment were 100% in group I, 90% in group II, 32% in group III and 10% in group IV. The serum biochemical findings, tissue ATP levels and hepatic blood flow were significantly improved in groups I and II compared with that in groups III and IV. The level of HPT in group I was the highest among the experimental groups. On histological findings, inflammatory changes were observed in group I and focal necrosis in group II while liver degeneration with massive necrosis was seen in groups III and IV. In conclusion, acute liver failure induced by D-Gal was apparently protected by the administration of DBcAMP. PMID- 2549364 TI - Isolation and characterization of the phosphoglucose isomerase gene from Escherichia coli. AB - The nucleotide sequence of the gene encoding the glycolytic enzyme phosphoglucose isomerase (PGI) from Escherichia coli is presented. The gene encodes a polypeptide of 549 amino acids. The transcriptional start point of the gene was determined and found to lie within a consensus promoter region. The amino acid sequence derived from the E. coli PGI gene can be aligned without insertions or deletions to the predicted amino acid sequence of a nuclear-encoded chloroplast isozyme of PGI from a higher plant, and the two sequences have a similarity of 87.6%. The amino acid sequence similarity between E. coli and that predicted from cDNA sequences for mouse and pig PGI is approximately 65%. PMID- 2549365 TI - Isolation and characterisation of transposon-induced mutants of Erwinia carotovora subsp. atroseptica exhibiting reduced virulence. AB - The blackleg pathogen Erwinia carotovora subsp. atroseptica (Eca) causes an economically important disease of potatoes. We selected a genetically amenable Eca strain for the genetic analysis of virulence. Tn5 mutagenesis was used to generate nine mutants which exhibited reduced virulence (Rvi-) of strain SCRI1043. Following physiological characterisation, mutants were divided into three classes: (1) auxotrophs; (2) extracellular enzyme mutants; and (3) a growth rate mutant. The isolation of these Rvi- mutants has allowed us to consider some factors that affect Eca virulence. PMID- 2549366 TI - Escherichia coli parA is an allele of the gyrB gene. AB - A thermosensitive (ts) parA mutant, MFT110, of Escherichia coli carried at least two ts mutations. The major ts defect, resulting from a mutation mapped originally at 95 min and complemented by pLC8-47, was most probably due to psd. A plasmid carrying the 1.6 kb BamHI-PvuII fragment recloned from pLC8-47 complemented the major ts mutation in MFT110 and psd(ts) in two mutants, but did not correct the Par phenotype of MFT110. The second ts mutation was salt repairable and mapped at 83 min close to recF and tnaA. This mutation was linked with the Par phenotype as shown unambiguously by 4',6-diamidino-2-phenylindole stained nucleoids in parA mutant cells with the W3110 genetic background. Both salt-repairable ts and Par traits were corrected concomitantly by a plasmid carrying the chromosomal region solely for the gyrB gene. This strongly suggests that parA is an allele of gyrB. PMID- 2549367 TI - Cloning of a gene involved in cellulose biosynthesis in Acetobacter xylinum: complementation of cellulose-negative mutants by the UDPG pyrophosphorylase structural gene. AB - Three cellulose-negative (Cel-) mutants of Acetobacter xylinum strain ATCC 23768 were complemented by a cloned 2.8 kb DNA fragment from the wild type. Biochemical analysis of the mutants showed that they were deficient in the enzyme uridine 5' diphosphoglucose (UDPG) pyrophosphorylase. The analysis also showed that the mutants could synthesize beta(1-4)-glucan in vitro from UDPG, but not in vivo from glucose. This result was expected, since UDPG is known to be the precursor for cellulose synthesis in A. xylinum. In order to analyze the function of the cloned gene in more detail, its biological activity in Escherichia coli was studied. These experiments showed that the cloned fragment could be used to complement an E. coli mutant deficient in the structural gene for UDPG pyrophosphorylase. It is therefore clear that the cloned fragment must contain this gene from A. xylinum. This is to our knowledge the first example of the cloning of a gene with a known function in cellulose biosynthesis from any organism, and we suggest the gene be designated celA. PMID- 2549368 TI - Induced transposition of Ds by a stable Ac in crosses of transgenic tobacco plants. AB - A two component maize transposon system comprising a stable but trans-active Ac element and a cis responsive Ds element has been established in transgenic tobacco. The development of this system is desirable for the realization of a gene tagging strategy employing these elements in heterologous plant species. Transgenic tobacco with a single transposed Ac element (Ac-18) which has sustained a 4 bp terminal deletion has been identified. Transposase activity of the stable Ac-18 was demonstrated in a tissue culture assay as well as in transgenic plants. When plants containing Ac-18 were crossed with transgenic plants carrying a Ds element, 25%-50% of the F1 progeny showed trans-activation of Ds transposition. Analysis of DNA of several F1 plants showed that each plant displayed a unique pattern of Ds transposition to new chromosomal sites. PMID- 2549369 TI - Efficient transformation of Arabidopsis thaliana using direct gene transfer to protoplasts. AB - Direct gene transfer has proved to be an efficient transformation method for Arabidopsis thaliana, a member of the Brassicaceae. Transgenic Arabidopsis plants resistant to hygromycin B have been regenerated from mesophyll protoplasts treated with polyethylene glycol and plasmid DNA carrying the hygromycin phosphotransferase (HPT) gene under the control of the 35 S promoter of cauliflower mosaic virus. The transformation procedure reproducibly yields transformants at frequencies of approximately 1 x 10(-4) (based on the number of protoplasts treated) or 5% (based on the number of regenerating calli). DNA from plants regenerated from hygromycin resistant colonies was analysed by Southern blot hybridization demonstrating that the foreign gene is stably integrated into the plant chromosome. Genetic analysis of several hygromycin resistant plants showed that the HPT gene is transmitted to the progeny. Transformation experiments performed with a selectable and a non-selectable gene on separate plasmids resulted in a co-transformation rate of functionally active copies in about 25% of the transformants analysed. Hence this approach can be used to introduce non-selectable genes into the Arabidopsis genome. PMID- 2549370 TI - Structure of the multigene family of MAL loci in Saccharomyces. AB - Multigene families are a ubiquitous feature of eukaryotes; however, their presence in Saccharomyces is more limited. The MAL multigene family is comprised of five unlinked loci, MAL1, MAL2, MAL3, MAL4 and MAL6, any one of which is sufficient for yeast to metabolize maltose. A cloned MAL6 locus was used as a probe to facilitate the cloning of the other four functional loci as well as two partially active alleles of MAL1. Each locus could be characterized as a cluster of three genes, MALR (regulatory), MALT (maltose transport or permease) and MALS (structural or maltase), encoded by a total of about 7 kb of DNA; however, homologous sequences at each locus extend beyond the coding regions. Our results indicate that there is extensive homology among the MAL loci, especially within their maltase genes. The greatest sequence diversity occurs in their regulatory gene regions. Southern cross analyses of the cloned MAL loci indicate a single duplication of the MAL6R-homologous sequences upstream of the MAL6R gene as well as an extensive duplication of more than 10 kb at the MAL3 locus. The large repeat at the MAL3 locus results in the presence of four copies of MAL3R homologous sequences and two copies of MAL3T-homologous sequences at that locus. Two naturally occurring inactive alleles of MAL1 show a deletion or divergence of their MALR sequences. The significance of these repeats in the evolution of the MAL multigene family is discussed. PMID- 2549372 TI - On the evolution of Tn21-like multiresistance transposons: sequence analysis of the gene (aacC1) for gentamicin acetyltransferase-3-I(AAC(3)-I), another member of the Tn21-based expression cassette. AB - The aminoglycoside-3-O-acetyltransferase-I gene (aacC1) from R plasmids of two incompatibility groups (R1033 [Tn1696], and R135) was cloned and sequenced. In the case of R1033, it was shown that the aacC gene is coded by a precise insertion of 833 bp between the aadA promoter and its structural gene in a Tn21 related transposon (Tn1696). This insertion occurs at the same target sequence as that of the OXA-1 beta-lactamase gene insertion in Tn2603. Upstream of the aacC gene, we found an open reading frame (ORF) which is probably implicated in the site-specific recombinational events involved in the evolution of this family of genetic elements. These results provide additional confirmation of the role of Tn21 elements as naturally occurring interspecific transposition and expression cassettes. PMID- 2549371 TI - The Escherichia coli dam gene is expressed as a distal gene of a new operon. AB - DNA containing the Escherichia coli dam gene and sequences upstream from this gene were cloned from the Clarke-Carbon plasmids pLC29-47 and pLC13-42. Promoter activity was localized using pKO expression vectors and galactokinase assays to two regions, one 1650-2100 bp and the other beyond 2400 bp upstream of the dam gene. No promoter activity was detected immediately in front of this gene; plasmid pDam118, from which the nucleotide sequence of the dam gene was determined, is shown to contain the pBR322 promoter for the primer RNA from the pBR322 rep region present on a 76 bp Sau3A fragment inserted upstream of the dam gene in the correct orientation for dam expression. The nucleotide sequence upstream of dam has been determined. An open reading frame (ORF) is present between the nearest promoter region and the dam gene. Codon usage and base frequency analysis indicate that this is expressed as a protein of predicted size 46 kDa. A protein of size close to 46 kDa is expressed from this region, detected using minicell analysis. No function has been determined for this protein, and no significant homology exist between it and sequences in the PIR protein or GenBank DNA databases. This unidentified reading frame (URF) is termed urf-74.3, since it is an URF located at 74.3 min on the E. coli chromosome. Sequence comparisons between the regions upstream of urf-74.3 and the aroB gene show that the aroB gene is located immediately upstream of urf-74.3, and that the promoter activity nearest to dam is found within the aroB structural gene. This activity is relatively weak (about 15% of that of the E. coli gal operon promoter). The promoter activity detected beyond 2400 bp upstream of dam is likely to be that of the aroB gene, and is 3 to 4 times stronger than that found within the aroB gene. Three potential DnaA binding sites, each with homology of 8 of 9 bp, are present, two in the aroB promoter region and one just upstream of the dam gene. Expression through the site adjacent to the dam gene is enhanced 2- to 4-fold in dnaA mutants at 38 degrees C. Restriction site comparisons map these regions precisely on the Clarke-Carbon plasmids pLC13-42 and pLC29-47, and show that the E. coli ponA (mrcA) gene resides about 6 kb upstream of aroB. PMID- 2549374 TI - Iron-hydroxamate transport into Escherichia coli K12: localization of FhuD in the periplasm and of FhuB in the cytoplasmic membrane. AB - The fhuB, fhuC and fhuD genes encode proteins which catalyze transport of iron(III)-hydroxamate compounds from the periplasm into the cytoplasm of Escherichia coli. The fhuB, C, D genes were cloned downstream of a strong phage T7 promoter and transcribed by T7 RNA polymerase. The overexpressed FhuD protein appeared in two forms of 31 and 28 kDa and was released upon conversion of vegetative cells into spheroplasts, suggesting synthesis of FhuD as a precursor and export into the periplasm. The very hydrophobic FhuB protein was found in the cytoplasmic membrane. These properties, together with the previously found homologies in the FhuC protein to ATP-binding proteins, display the characteristics of a periplasmic binding protein dependent transport system across the cytoplasmic membrane. The molecular weight of FhuB and the sequence of fhuC, as previously published by us, was confirmed. FhuB exhibited double the size of most hydrophobic proteins of such systems and showed homology between the amino- and carboxy-terminal halves of the protein, indicating duplication of an original gene and subsequent fusion of the two DNA fragments. PMID- 2549373 TI - Genetic deletions between directly repeated sequences in bacteriophage T7. AB - DNA sequence analysis of genetic deletions in bacteriophage T7 has shown that these chromosomal rearrangements frequently occur between directly repeated DNA sequences. To study this type of spontaneous deletion in more quantitative detail synthetic fragments of DNA, made by hybridizing two complementary oligonucleotides, were introduced into the non-essential T7 gene 1.3 which codes for T7 DNA ligase. This insert blocked synthesis of functional ligase and made the phage that carried an insert unable to form plaques on a host strain deficient in bacterial ligase. The sequence of the insert was designed so that after it is put into the T7 genome the insert is bracketed by direct repeats. Perfect deletion of the insert between the directly repeated sequences results in a wild-type phage. It was found that these deletion events are highly sensitive to the length of the direct repeats at their ends. In the case of 5 bp direct repeats excision from the genome occurred at a frequency of less than 10(-10), while this value for an almost identical insert bracketed by 10 bp direct repeats was approximately 10(-6). The deletion events were independent of a host recA mutation. PMID- 2549375 TI - Nucleotide sequences from the colicin E5, E6 and E9 operons: presence of a degenerate transposon-like structure in the ColE9-J plasmid. AB - The nucleotide sequences of 1288 bp of plasmid ColE5-099, 1609 bp of ColE6-CT14 and 2099 bp of ColE9-J were determined. These sequences encompass the structural genes for the C-terminal receptor-binding and nuclease domains of colicins E5, E6 and E9, their cis- or trans-acting immunity proteins and four lysis proteins including an atypical one of non-lipoprotein nature (Lys) present in the ColE9-J plasmid. The ColE6 gene organisation, in the order col-imm-E8imm-lys, is identical to that found in the previously described double-immunity gene system of ColE3-CA38 (an RNase producer). The corresponding genes in the two plasmids are 87%-94% homologous. In ColE9-J, the genes are organised as col-imm-lys-E5imm lys. The E9 col-imm gene pair is homologous to the colicin E2-P9 type (a DNase producer). Downstream from E9imm is an E5imm (designated E5imm[E9]) which is trans-acting. Neither the predicted structures of E5Imm[E9] nor the cis-acting Imm resident in the ColE5-099 plasmid which differs by a single amino acid shows any resemblance to other immunity structures which have been sequenced. Furthermore, the E5col sequences differ from those predicted previously for other colicins except for the conserved btuB-specified receptor-binding domain. A novel 205 nucleotide long insertion sequence is found in the ColE9-J plasmid. This insertion sequence, which we named ISE9, has features reminiscent of the degenerate transposon IS101 previously found in plasmid pSC101. One effect of ISE9 is the presence of the atypical lysis gene, lys.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549376 TI - Use of transformation to make targeted sequence alterations at the am (GDH) locus of Neurospora. AB - Specific in vitro-generated insertion, replacement, and deletion mutations have been integrated near the chromosomal locus of am (NADP-specific glutamate dehydrogenase) of Neurospora crassa. Two approaches have been successful. One approach used am+-containing vectors capable of integrating at any site in the genome. This technique was used to introduce a specific 700 bp insertion near the am locus and to replace chromosomal sequences near am with plasmid DNA. Efficiency was low, however, and many transformants had to be screened to find the desired alterations among the ectopic insertions unless the incoming DNA had a large region of homology with the am region. A second approach increased the efficiency by using vectors containing a truncated am gene, so that prototrophs could arise only by homologous recombination. Overall transformation frequency was reduced relative to the first method, but a large fraction of the transformations involved specific alterations of the am region. PMID- 2549377 TI - Micrococcus luteus homolog of the Escherichia coli uvrA gene: identification of a mutation in the UV-sensitive mutant DB7. AB - Restriction fragments of Micrococcus luteus DNA containing the gene affected by a mutation in the UV-sensitive mutant DB7 were cloned both from the wild type and from the mutant in an Escherichia coli host-vector system. The wild-type fragment was able to reverse the multiple sensitivity of the mutant to UV, mitomycin C, and 4-nitroquinoline 1-oxide by a one-step transformation. Determination of the nucleotide sequences revealed a potential open reading frame coding for a protein of 992 (tentative) amino acid residues, within which the DB7 mutation was identified as a CG-to-TA transition causing a translation termination. The putative product of the open reading frame shares an extensive amino acid sequence homology with the E. coli UvrA protein comprising 940 residues. The homology extends over the greater part of both polypeptides except for two extra sequences of 31 and 24 amino acid residues located at the amino-terminal and in the interior, respectively, of the M. luteus protein. In the homologous region, 56.7% and 16.7% of the 933 pairs of the aligned amino acids were accounted for by conserved residues and conservative substitutions, respectively. These results indicate that the gene defined by the mutation in DB7 represents a homolog of the E. coli uvrA gene. Hence, it has to be concluded that DB7, known for its deficiency in UV endonuclease (pyrimidine dimer DNA glycosylase/apurinicapyrimidinic endonuclease) activity, is a double mutant which is also defective in an enzyme complex similar to the E. coli UvrABC excinuclease. PMID- 2549378 TI - Transport across the outer membrane of Escherichia coli K12 via the FhuA receptor is regulated by the TonB protein of the cytoplasmic membrane. AB - Point mutations in the "TonB box" of fhuA were suppressed by point mutations in the tonB gene, suggesting both a functional and physical interaction between the FhuA receptor protein in the outer membrane and the TonB protein in the cytoplasmic membrane of Escherichia coli K12. Mutations in fhuA were classified into four types according to the extent by which they impaired mutant cells in their growth on ferrichrome as sole iron source and in their sensitivity to the antibiotic albomycin and to colicin M. The tonB mutation with a glutamine to leucine replacement at position 165 was less efficient in restoring the FhuA functions than the glutamine to lysine exchange at the same position. The better the coupling between FhuA and TonB the poorer was the inhibition of phage T1 binding to FhuA by ferrichrome. A working model is proposed in which the TonB protein assumes different conformations in response to the energized state of the cytoplasmic membrane and thereby allosterically regulates the activity of the FhuA receptor. This model implies an intermembrane coupling between two proteins in adjacent membranes. PMID- 2549379 TI - Regulation of repressor and early gene expression in Mu-like transposable bacteriophage D108. AB - The temperate, transposable bacteriophages D108 and Mu are highly homologous, but differ in their lef-end regulatory regions. We have previously cloned the gene encoding the D108 thermo-sensitive (cts) repressor under the control of the lactUV5 promoter. In this work, we report that crude protein extracts containing highly-expressed D108 repressor protect a 77 bp region of DNA, located between 863 bp and 940 bp from the D108 lef--end, from both exonuclease III and DNase I hydrolysis. Nucleotide sequence analysis of this region reveals that is also contains DNA sequences homologous to the consensus DNA-binding site of the Escherichia coli protein, Integration Host Factor (IHF). Crude protein extracts containing highly-expressed IHF specifically bind to, and retard the migration of, DNA fragments containing the D108 regulatory region, and the DNA sequence which IHF protects from DNase I cleave lies directly within the D108 repressor binding region. There are two apparent repressor-specific S1 nuclease-resistant RNA suggests that transcription from the early region promoter, Pe may initiate at or about 1000 bp from the left-end of the D108 genome. Thus though, D108 and Mu utilize three analogous proteins (repressor, ner, and IHF) and the same apparent promoters for early gene regulation and the lytic/lysogenic decision, the organization of these regulatory components is apparently different, suggesting different mechanisms of control of gene expression. PMID- 2549380 TI - An unusual wheat insertion sequence (WIS1) lies upstream of an alpha-amylase gene in hexaploid wheat, and carries a "minisatellite" array. AB - Comparison of the 5' flanking regions of three alpha-amylase genes from chromosome 6B of hexaploid wheat by heteroduplex and sequence analysis revealed the presence of a 1.6 kb stem-loop insertion sequence (WIS1) in one of them. Polymorphism among hexaploid wheat varieties suggests the relatively recent insertion/excision of this sequence from its present position. The complete sequence of the stem-loop insertion shows that it has many of the features found in transposable elements, including target site duplication and terminal inverted repeats. One unusual feature is a tandem array of direct repeats comprising a wheat "minisatellite" sequence. Both the insertion sequence and the minisatellite are found at multiple locations in the wheat genome, but the functional significance of their association in WIS1 is unknown. The minisatellite arrays share a common core structure, and long arrays are polymorphic between different hexaploid varieties. PMID- 2549381 TI - The bz-rcy allele of the Cy transposable element system of Zea mays contains a Mu like element insertion. AB - The receptive component of the Cy transposable element system (rcy:Mu7) at the Bz locus of Zea mays L. is 2.2 kb and has long terminal inverted repeats. The insertion is flanked by a 9 bp duplication. In the presence of an autonomous Cy element in the genome, rcy:Mu7 is excised from bz-rcy in a manner consistent with a model suggested previously. The termini of rcy:Mu7 have 85% sequence similarity with the Mu1 element of Z. mays. This is consistent with the observation that Mu1 can behave genetically like a receptive component of the Cy system. PMID- 2549382 TI - Beta 2-adrenergic receptor regulation after transfection into a cell line deficient in the cAMP-dependent protein kinase. AB - A mouse beta 2-adrenergic receptor (beta 2AR) DNA clone was transfected and expressed in a mouse adrenocortical tumor cell line (Kin8) that lacks both beta 2AR and cAMP-dependent protein kinase (PKA). The receptor displayed a characteristic beta 2AR agonist binding profile that was similar to that observed in beta 2AR-transfected PKA+ mouse adrenocortical tumor cells (Y1). Isoproterenol treatment of beta 2AR-transfected Kin8 and Y1 cells resulted in a rapid loss of surface beta 2AR, as determined by the binding of the hydrophilic beta 2AR radioligand [3H]CGP 12177 [( 3H]CGP), followed by a decrease in adenylate cyclase activity. Sequestration of beta 2AR in Kin8 cells was beta 2AR agonist specific, temperature dependent, and rapidly reversible. Repeated treatment and recovery from isoproterenol incubation resulted in a cycling of surface [3H]CGP binding. The reappearance of [3H]CGP binding following short isoproterenol treatment was not affected by cycloheximide treatment of the cells. Prolonged incubation of beta 2AR-transfected Kin8 cells with isoproterenol resulted in the down regulation of beta 2AR protein without a change in beta 2AR mRNA levels. Polysome profiles of control and down-regulated cells revealed that translation of beta 2AR mRNA is inefficient and does not change upon prolonged agonist treatment. Protein synthesis was required to reverse the down-regulation of beta 2AR. These results indicate that neither sequestration nor down-regulation of beta 2AR depends on PKA. PMID- 2549383 TI - Multiple opioid receptors: ligand selectivity profiles and binding site signatures. AB - Methods are described for studying mu, delta, and kappa opioid binding sites, each without interference from the others. A large array of ligands has been characterized by ligand selectivity profiles, graphic depictions of affinities and selectivities. Binding site signatures have been derived, which uniquely describe each of the three types of sites. The mu, delta, and kappa binding sites have interesting common features and distinctive differences. PMID- 2549384 TI - Activation of sodium channels and inhibition of [3H]batrachotoxinin A-20-alpha benzoate binding by an N-alkylamide neurotoxin. AB - BTG 502 [(2E,4E)-N-(1,2-dimethyl)-propyl-6-(5-bromonaphth-2-yl)-hexa -2,4- dienamide], a synthetic analog of insecticidal amides isolated from Piper species, stimulated 22Na+ uptake into mouse brain synaptoneurosomes in the presence of saturating concentrations of Leiurus quinquestriatus venom but had no effect on sodium uptake in the absence of venom. In the presence of Leiurus venom, half-maximal stimulation was achieved at a BTG 502 concentration of 1.7 microM, whereas maximal stimulation (2.3-fold greater than nonspecific uptake) was observed at 50 microM. In the absence of other modifiers, BTG 502 inhibited batrachotoxin (BTX)-dependent sodium uptake, producing 50% inhibition at 2 microM. In the presence of Leiurus venom, BTG 502 was a partial inhibitor of BTX dependent 22Na+ uptake, producing half-maximal inhibition at 1.5 microM. The levels of residual BTX-dependent sodium uptake and maximal BTG 502-dependent sodium uptake measured in the presence of Leiurus venom were identical. BTG 502 inhibited the specific binding of [3H]batrachotoxinin A-20-alpha-benzoate (BTX-B) to the activator recognition site (site 2) of sodium channels in these preparations, producing half-maximal inhibition at 2 microM and maximal inhibition at 30 microM. Equilibrium analysis showed that BTG 502 was an apparent competitive inhibitor of [3H]BTX-B binding, producing a concentration-dependent decrease in the affinity of sodium channels for this ligand without affecting binding capacity. Kinetic analysis demonstrated that BTG 502 slowed the rate of formation of the ligand-receptor complex but did not alter the rate of dissociation of this complex. The effects of BTG 502 on 22Na+ uptake and [3H]BTX B binding are consistent with the action of this compound as an antagonist at the activator recognition site of the voltage-sensitive sodium channel in the absence of Leiurus venom and as a partial agonist at this site in the presence of Leiurus venom. These results suggest that the N-alkylamides represent a novel chemical class of neurotoxins that act at site 2 of the sodium channel. PMID- 2549385 TI - Phosphorylation of 2',3'-dideoxyinosine by cytosolic 5'-nucleotidase of human lymphoid cells. AB - 2',3'-Dideoxyinosine (ddlno) is a potent and selective inhibitor of human immunodeficiency virus in human lymphoid cells and monocytes/macrophages. Earlier studies [J. Biol. Chem. 263:15354 (1988)] showed that anabolism of ddlno in human lymphoid cells is mediated via an initial step of phosphorylation and subsequent amination to dideoxy-AMP via adenylosuccinate synthetase/lyase. Evidence was obtained that neither adenosine kinase nor deoxycytidine kinase is involved in the phosphorylation of this compound in human lymphoid cells. We now find that, in the presence of MgCl2, KCl, and inosine-5'-monophosphate as phosphate donor, purified cytosolic 5'-nucleotidase catalyzed the phosphorylation of ddlno. Although not phosphate donors, ATP, diadenosine tetraphosphate, and glycerate-2,3 bisphosphate stimulate this phosphorylation by the nucleotidase 4-5-fold. In addition to ddlno, the antiviral nucleoside analogs 2',3'-dideoxyguanosine and carbovir were substrates for this enzyme. The relative phosphorylation of these compounds varied with the concentration of the phosphate donor IMP. Approximate Km values of the nucleotidase for inosine, ddlno, dideoxyguanosine, and carbovir were, respectively, 3.4, 0.5, 0.9, and 1.7 mM. Although the substrate activity of dideoxynucleosides is inefficient, it appears likely that this nucleotidase is responsible for the metabolism of these compounds to their active nucleotides, yielding antiviral activity in human lymphoid cells. PMID- 2549386 TI - Mechanism of selective inhibition of varicella zoster virus replication by 1-beta D-arabinofuranosyl-E-5-(2-bromovinyl)uracil. AB - To investigate the mechanism of action of 1-beta-D-arabinofuranosyl-(E)-5-(2 bromovinyl)uracil (BV-araU) on varicella zoster virus (VZV) replication, we examined the metabolism of the drug in VZV-infected cells using 14C-labeled BV araU. [14C]BV-araU was taken up by the cells infected with thymidine kinase positive (TK+)VZV, but not so much by TK- VZV-infected or mock infected cells. Most of the radioactivity in TK+ VZV-infected cells that were incubated with [14C]BV-araU was recovered from their acid-soluble fraction, and little from their acid-insoluble fraction. By high performance liquid chromatographic assay of the acid-soluble fraction, it was proved that BV-araU was metabolized to its 5'-monophosphate, diphosphate, and triphosphate only in TK+ VZV-infected cells. The radioactivity was not detected in VZV nucleocapsids or in VZV DNA and cellular DNA isolated from TK+ VZV-infected cells, even if BV-araU was added at a 1000 times higher concentration than the 50% inhibitory dose for VZV replication in vitro. Furthermore, it was enzymatically proved that [14C]BV-araU was selectively and effectively phosphorylated to BV-araU monophosphate by VZV TK and that affinity of BV-araU triphosphate for VZV DNA polymerase was the quite strong. From these results, it can be concluded that marked inhibition of VZV replication by BV-araU is due to selective phosphorylation of BV-araU in the TK+ VZV-infected cells and strong inhibition of VZV DNA synthesis by BV-araU triphosphate, without detectable incorporation into VZV DNA. PMID- 2549387 TI - Defining the role of protein kinase C in epinephrine- and bradykinin-stimulated arachidonic acid metabolism in Madin-Darby canine kidney cells. AB - Madin-Darby canine kidney cells (MDCK) are known to release free arachidonic acid and arachidonic acid metabolites (AA) in response to tumor-promoting phorbol esters, such as tetradecanoyl phorbol-13-acetate, and to agonists active at alpha 1-adrenergic and bradykinin B2 receptors. These experiments were conducted to define the role of Ca2+/phospholipid-dependent protein kinase (protein kinase C) activation in the stimulation of AA release, in the clonal isolate cell line MDCK D1, by use of three inhibitors of protein kinase C, sphingosine, 1-(5 isoquinolinesulfonyl)-2-methylpiperazine (H-7), and staurosporine. We found that alpha 1-adrenergic- and bradykinin-stimulated [3H]AA release can be distinguished by differential dependence on protein kinase C; epinephrine-stimulated release was more dependent on protein kinase C activation than was bradykinin-stimulated release. The inhibition of bradykinin-stimulated AA release by sphingosine (20.2 +/- 6.1%) was substantially less than the inhibition observed for tetradecanoyl phorbol-13-acetate- (67.2 +/- 5.5%) and epinephrine-stimulated release (50.2 +/- 9.2%). These findings were confirmed by results using H-7 and staurosporine. The relative independence of bradykinin-stimulated AA release of protein kinase C was also demonstrated by the inability of phorbol ester-induced down-regulation of protein kinase C to eliminate bradykinin-stimulated AA release. The inhibition of alpha 1-adrenergic receptor-mediated AA release by sphingosine, H-7, and staurosporine was not due to a change in receptor number or affinity. Analysis of the products comprising [3H]AA release indicated that treatment with sphingosine did not change the composition of the released AA (34-48% prostaglandin E2, 17 27% free arachidonic acid, and 25-51% unidentified metabolites). These results indicate that two different types of hormone receptors in the same cell type can promote AA release by mechanisms that differ in their dependence on protein kinase C. The protein kinase C-dependent mechanism may represent protein kinase C mediated activation of phospholipase A2. PMID- 2549388 TI - Radiation inactivation reveals discrete cation binding sites that modulate dihydropyridine binding sites. AB - In low ionic strength buffer (5 mM Tris.HCl), the binding of [3H] nitrendipine to dihydropyridine calcium antagonist binding sites of mouse forebrain membranes is increased by both Na+ and Ca2+. Radiation inactivation was used to determine the target size of [3H]nitrendipine binding sites in 5 mM Tris.HCl buffer, in the presence and absence of these cations. After irradiation, [3H] nitrendipine binding in buffer with or without Na+ was diminished, due to a loss of binding sites and also to an increase in Kd. After accounting for radiation effects on the dissociation constant, the target size for the nitrendipine binding site in buffer was 160-170 kDa and was 170-180 kDa in the presence of sodium. In the presence of calcium ions, [3H]nitrendipine binding showed no radiation effects on Kd and yielded a target size of 150-170 kDa. These findings suggest, as in the case of opioid receptors, the presence of high molecular weight membrane components that modulate cation-induced alterations in radioligand binding to dihydropyridine binding sites. PMID- 2549389 TI - Regulation of the ATP-dependent calcium uptake activity of heart sarcolemmal vesicles by endogenous cytosolic proteins. AB - In a previous study we described the inhibitory action of a cytosolic protein fraction from heart muscle on ATP-dependent Ca2+ uptake by sarcoplasmic reticulum; further, this inhibition was shown to be blocked by an inhibitor antagonist, also derived from the cytosol (Narayanan et al. Biochim Biophys Acta 735: 53-66, 1983). The present study examined the effects of the endogenous cytosolic Ca2+ transport inhibitor and its antagonist on ATP-dependent Ca2+ uptake by sarcolemmal vesicles isolated from rat and canine heart. The cytosolic inhibitor caused strong inhibition (up to 97%) of Ca2+ uptake by sarcolemma (SL); this inhibition could be reversed by the cytosolic inhibitor antagonist. Studies on the characteristics of inhibition revealed the following: a) Inhibition was dependent on the concentration of the inhibitor (50% inhibition with approximately 80 micrograms inhibitor protein). b) The inhibitor reduced the velocity of Ca2+ uptake without appreciably influencing the apparent affinity of the transport system for Ca2+ but caused greater than 2-fold decrease in its apparent affinity for ATP. c) The rates of unidirectional passive Ca2+ release from actively Ca2+ loaded SL vesicles were not altered by low concentrations of the inhibitor (less than 100 micrograms/ml) which were effective in producing marked inhibition of Ca2+ uptake; at higher concentrations (greater than 100 micrograms/ml), the inhibitor caused increase in the rates of passive Ca2+ release. These findings demonstrate that the activity of the ATP-driven Ca2+ pump of cardiac SL can be regulated in vitro by endogenous cytosolic proteins. PMID- 2549390 TI - Characterization of muscarinic acetylcholine receptors on intact neuroblastoma x glioma NG108-15 cell upon induced differentiation. AB - Studies have established that major increases in muscarinic acetylcholine receptor (mAchR) binding in the brain appear to coincide with synaptogenesis. The neuroblastoma x glioma hybrid NG108-15 cell line has been demonstrated to possess numerous functional characteristics of intact neurons, including synapse formation with myotubes. The present study examines and characterizes the mAchR on the hybrid NG108-15 cells during differentiation, induced by 1 mM dBcAMP. Specific binding of [3H]-QNB for differentiated cells increases gradually to a final level of 130% (P less than 0.05) over the control undifferentiated cells during the first 24 hr of incubation. Further, this increase of receptor sites appears to correlate proportionately to the degree of neurite extension of the differentiating cells. The dissociation rate constant at equilibrium (Kd) and maximum binding capacity (Bmax) have been determined to be 5.6 nM and 920 fmol/10(6) cells, respectively, for differentiated cells, and 4.4 nM and 400 fmol/10(6) cells, respectively, for undifferentiated cells. Computer analyses of the data obtained from saturation experiments reveal a single class of binding sites for [3H]-QNB on both differentiated and undifferentiated cells. The Hill plot analysis of the QNB-binding indicates a Hill coefficient (nH) of 1.0 and 0.91 for differentiated and undifferentiated cells, respectively, suggesting the unity of receptor sites with no co-operativity. Our results depict that increases of mAchRs on intact cells correlate with the degree of cellular differentiation. PMID- 2549393 TI - [Modification of human DNA polymerase alpha by N-acetylimidazole]. AB - The modification of tyrosine residues of the human placenta DNA-polymerase alpha by N-acetylimidazole was investigated. The poly(dT)-template and the r(pA)10 primer a each added separately or simultaneously do not influence the rate of enzyme inactivation. In the presence of poly(dT)-r(pA)10 no effect of dCTP and dTTP (noncomplementary to template) and of dAMP and dADP (complementary to template) on the rate and the level of the enzyme inactivation was found. However dATP revealed practically complete protection. Orthophosphate, pyrophosphate each taken separately do not influence the rate of enzyme inactivation with this reagent. The presence of dADP with either ortho- or pyrophosphate, or dAMP with the one of these ligands leads to half protective action in comparison with dATP. Imidazolides of phosphonoacetic acid and 5'-adenylyl++ 1(phosphonoacetic acid) do not inactivate DNA-polymerase alpha from human placenta and the Klenov fragment of DNA-polymerase I from E. coli. All data obtained allow to suggest that the tyrosine residue in the dNTP binding site of DNA-polymerase reveals stacking with the nucleotide only if dNTP is complementary to the template. PMID- 2549391 TI - Identification and partial characterization of a latent ATP, Mg-dependent protein phosphatase in rabbit skeletal muscle cytosol. AB - Fractionation of rabbit skeletal muscle cytosol on Aminohexyl-Sepharose has resulted in the identification of a latent ATP, Mg-dependent protein phosphatase whose catalytic subunit is in the active conformation, but is inhibited by the presence of more than one modulator unit. The partially purified enzyme is converted to an inactive, kinase FA-dependent form upon incubation at 30 degrees C unless modulator-specific polyclonal antibodies are added to the preparation. The immunoglobulins also relieve the inhibition which is responsible for the low basal phosphatase activity of the enzyme, and they counteract all of the heat stable inhibitor activity present in the preparation. Addition of free catalytic subunit abolishes the inhibition of the latent enzyme in a dose-dependent way, but cannot prevent the inactivation process. The inactivated phosphatase and the original latent enzyme exhibit the same apparent Mr in sucrose density-gradient centrifugation (70,000) and in gel filtration (110,000). PMID- 2549392 TI - Physiological relevance of the changing subunit composition and regulatory properties of the 6-phosphofructo-1-kinase isozyme pools during heart and muscle development. AB - During postnatal development, the subunit compositions of the 6-phosphofructo-1 kinase isozyme pools of heart and skeletal muscle are known to change. The isozyme pools from fetal muscle were composed of the L-type (60%), and M-type (36%) and C-type (4%) subunits and the isozymes from fetal and early neonatal heart contain nearly equal amounts of all three subunits. During postnatal development of both tissues, the proportion of the M-type subunit increases until it is the only type present in adult muscle and the major subunit in adult heart (75%). The isozyme pool from fetal muscle exhibit a decreased affinity for fructose-6-P and a greater susceptibility to ATP inhibition compared to the M rich isozymes which are subsequently present. The isozyme pools from fetal and early neonatal heart, if compared to the M-rich isozymes which are present later during heart development and to the fetal muscle isozymes, exhibited the least affinity for fructose-6-P and the greatest susceptibility to ATP inhibition. Comparison of the isozyme pools containing little or no C-type subunit with those from fetal and early neonatal heart clearly indicates that the presence of substantial levels of the C-type subunit imposed a decreased ability for fructose 2,6-P2 to both lower affinity for fructose-6-P and antagonize sensitivity to ATP inhibition. Although still not thoroughly appreciated, it appears that the changing nature of the isozyme pools in these tissues permits regulation of glucose metabolism in a manner which allows efficient utilization of nutritional opportunities and which adequately meets the energy requirements of each tissue at different stages of development. PMID- 2549395 TI - [Effect of camptothecin on the DNA-relaxing and DNA-cleavage activity of calf thymus topoisomerase I]. AB - Nucleotide sequences that are cleaved by calf thymus type I topoisomerase have been determined using cloned human Ha-ras and p53 genes. Localization and relative frequency of single-strand cleavages within these sequences were observed to change in the presence of the cytotoxic alkaloid camptothecin. PMID- 2549394 TI - [Cloning and comparative characteristics of genes coding two structurally distant delta-endotoxins of Bacillus thuringiensis var. galleriae and kurstaki]. AB - Representative total DNA libraries of Bac. thuringiensis var. kurstaki (strain Dipel) and galleriae (strain 11-67) have been constructed on the basis of phasmid vector lambda pSL5. Recombinant phasmid clones, carrying delta-endotoxin-coding genes of both subspecies have been isolated by means of immunoenzyme screening. Restriction mapping and partial nucleotide sequence determination have demonstrated that phasmid lambda pOC2, isolated from var. kurstaki DNA library, contains the complete delta-endotoxin-coding gene, identical to the one, described by Schnepf H.E. et al. J. Biol. Chem. 1985. V. 260. P. 6264. Recombinant phasmids lambda pOC10 and 11 have been shown to contain the full sized gene, coding delta-endotoxin of Bac. thuringiensis var. galleriae. The protein products of the cloned genes have been characterized by Western-blot analysis and bioassays. The absence of substantial homology of two genes, evidenced by Southern-blot hybridisation, correlates with sufficiently big differences in biological specificity of the corresponding proteins. This is in accordance with our previous data on N-terminal amino acid sequence determination of delta-endotoxins of those subspecies of Bac. thuringiensis. PMID- 2549397 TI - [Distribution of Alu repeats along the human genome: formation of clusters and features of insertion regions]. AB - The contextual analysis of nucleotide sequences of 22 Alu repeats arrangement regions in the human genome has been carried out and some of their peculiarities have been revealed. In particular, the occurrence of marked and statistical non random homology between the repeats and the regions of their integration has been shown. A mechanism of choosing the Alu repeats insertion regions in the genome has been suggested taking into account these peculiarities. Using a sample of the 80 human Alu repeats sequences peculiarities of these repeats location within the genome has been investigated. A tendency to the formation of Alu repeats clusters in various regions of the genome was revealed. A range of possible mechanisms on such Alu clusters emergence is considered. On the basis of the data obtained an "attraction" mechanism, according to which integration of Alu repeats into the definite region of the genome increases the insertion probability of other Alu repeats into the same region, are proposed. PMID- 2549396 TI - [DNA helix-coil transition in alkaline medium]. AB - Dna helix-coil transition in th alkaline medium was considered theoretically and experimentally. On the basis of the theory and experimental comparison the DNA double-stranded form deprotonation was revealed. PMID- 2549398 TI - [Molecular mechanisms of receptor-mediated endocytosis and their use in directed transport of biologically active compounds]. AB - The review considers the general and specific mechanisms of receptor-mediated endocytosis of various endogenic and exogenic bioactive compounds. The application of these mechanisms for developing of effective approaches to site directed transport of bioactive compound to certain target cells and tissues and ways of their employment in biology and medicine are discussed. PMID- 2549399 TI - [Molecular weight distribution of polymeric DNA-products, formed upon the action of matrix enzymes]. AB - Models of processive and distributive DNA synthesis and degradation catalysed by matrix enzymes were investigated. Distribution of polymer products dependent on the reaction model chosen, on the type of the matrix and on the enzyme-matrix initial concentration ratio were determined by methods of numerical modeling. Conditions were found where the scattering of the reaction polymer products was minimal. The homopolymer matrix choice of experimental condition may generate a distribution of product that obeys the Poisson distribution. Numerical investigations of polymerization (hydrolysis) processes showed that for a number of heteropolymer matrixes a distribution may exist with scattering much less than that for homopolymer matrixes. Conditions are found when processive and distributive models give different distributions of the reaction product. PMID- 2549400 TI - [Activity of topoisomerase I and endonucleases in cells transfected by a ras oncogene]. AB - Activities of nuclear endonucleases and topoisomerase I were measured in rat fibroblasts which were at the stages of tumor transformation: control embryonal fibroblasts--CEF; cells immortalised by transfection of S1A segment of SA7 adenovirus--REF-1; intermedius cells transfected once by EJras oncogene--REF-1EJ; and cells transformed after the second transfection by the same oncogene--REF 2EJ. The topoisomerase I and Ca2+, Mg2+-dependent endonuclease was most decreased at the stage of immortalised cells, and the intermedius stage (REF-1EJ) was characterized by the lower activity of Ca2+, Mg2+-dependent endonuclease. The highest activity of Mn2+-dependent endonuclease is seen in REF-2EJ cells. In model experiments the ability of Ca2+, Mg2+-dependent endonuclease to split non stochastically the EJras oncogene inserted into pBR322 plasmid was shown. The role of the investigated enzymes in the restriction of plasmid integration, cellular immortalisation and recombination of plasmids with chromosomes during cell transformation is discussed. PMID- 2549401 TI - [Differences in the distribution of proviruses in low- and highly- metastatic variants of hamster cells, transformed by Rous sarcoma virus]. AB - The structure of provirus in Syrian hamster cells, transformed by Rous sarcoma virus (RSV) varying in metastatic capability in vivo has been analysed. The original cell line and its low metastatic variants contain only one copy of the integrated RSV genome. The DNA of highly metastatic cell lines cloned from the same primary culture, contain an additional copy of provirus. This RSV copy in different highly metastatic variants has a similar integration site. PMID- 2549402 TI - [Analogs of pyrophosphate in a pyrophosphorolysis reaction catalyzed by DNA polymerases]. AB - The reaction of pyrophosphorolysis catalyzed by Escherichia coli DNA polymerase I Klenov fragment, calf thymus DNA polymerase alpha, rat liver DNA polymerase beta and AMV reverse transcriptase was studied. Some pyrophosphate (PPi) analogs were taken as low molecular weight substrates. It was shown that only imidodiphosphonic acid acted as the PPi substrate analog for the reactions catalyzed by DNA polymerases I and alpha, both imidodiphosphonic acid and methylenediphosphonic acid were active in the case of DNA polymerase beta and reverse transcriptase. Other analogs tested were neither nucleotide residue acceptors, nor inhibitors of the pyrophosphorolysis reaction with PPi. The abilities of some PPi analogs to inhibit the DNA elongation catalyzed by reverse transcriptase were investigated. The principles of specificity of low molecular substrates recognition by DNA polymerases and some problems concerning the mechanisms of DNA synthesis inhibition by PPi analogues are discussed. PMID- 2549403 TI - Interleukin-1 induction by lipopolysaccharides: structural requirements of the 3 deoxy-D-manno-2-octulosonic acid (KDO). AB - We previously showed the importance of the 3-deoxy-D-manno-2-octulosonic acid (KDO) residue in endotoxins (lipopolysaccharides, LPS) for the induction of the synthesis and release of interleukin-1 (IL-1) by human monocytes. We further investigated the effect of some structural variations within the KDO molecule on IL-1 production induced by LPS. Deamination of Bordetella pertussis LPS, followed by mild anhydrous acidic methanolysis released a hexasaccharide (fragment B'), which had a terminal methyl ketoside KDO residue with a methyl-esterified carboxyl group. This fragment was unable to induce IL-1 production by human monocytes. Fragment B' could be converted into an active hexasaccharide by de esterification (fragment B-OMe), but not by reduction of the methyl ester group. The KDO residues in the LPS of some bacterial species have been shown to be phosphorylated and we observed that these LPS were weak IL-1 inducers. Phosphorylated KDO present in Vibrio cholerae and B. pertussis LPS respond poorly in the thiobarbiturate assay (specific for KDO). However, if these LPS were dephosphorylated with aqueous hydrofluoric acid (HF) their KDO response in this assay was increased 5.4- to 2.6-fold, respectively. In parallel, the HF-treated LPS were more potent IL-1 inducers than untreated endotoxins. These data confirm that the KDO residue(s) present in all endotoxins play(s) a major role in the signal(s) leading to IL-1 production by human monocytes, and show that IL-1 induction by LPS (1) requires a free carboxyl group in the KDO and (2) is correlated with the degree of substitution of the KDO. PMID- 2549404 TI - A novel interferon-gamma regulated human melanoma-associated antigen, gp33-38, defined by monoclonal antibody Me14-D12. II. Molecular cloning of a genomic probe. AB - The human Me14-D12 antigen is a cell surface glycoprotein regulated by interferon gamma (IFN-gamma) on tumor cell lines of neuroectodermal origin. It consists of two non-convalently linked subunits with apparent mol. wt sizes of 33,000 and 38,000. Here we describe the molecular cloning of a genomic probe for the Me14 D12 gene using the gene transfer approach. Mouse Ltk- cells were stably cotransfected with human genomic DNA and the Herpes Simplex virus thymidine kinase (TK) gene. Primary and secondary transfectants expressing the Me14-D12 antigen were isolated after selection in HAT medium by repeated sorting on a fluorescence activated cell sorter (FACS). A recombinant phage harboring a 14.3 kb insert of human DNA was isolated from a genomic library made from a positive secondary transfectant cell line. A specific probe derived from the phage DNA insert allowed the identification of two mRNAs of 3.5 kb and 2.2 kb in primary and secondary L cell transfectants, as well as in human melanoma cell lines expressing the Me14-D12 antigen. The regulation of Me14-D12 antigen by INF-gamma was retained in the L cell transfectants and could be detected both at the level of protein and mRNA expression. PMID- 2549405 TI - Use of synthetic oligodeoxyribonucleotides for type-specific identification of coxsackie B viruses. AB - Synthetic oligodeoxyribonucleotides were used for type-specific identification of members of the coxsackie B virus group by nucleic acid hybridization. Two pairs of oligonucleotide chains were constructed based on nucleotide sequences in the VP1 regions of coxsackieviruses B3 and B4. Each labelled probe had a length of 24 nucleotides. The results showed that the oligonucleotide hybridized in a type specific manner when assayed with extracts from cells infected with all different coxsackie B viruses. A method based on similar principles may thus be used for enterovirus typing. PMID- 2549406 TI - Selective detection of human papilloma virus DNAs by specific synthetic DNA probes. AB - Specific oligodeoxynucleotide probes ranging from 20 to 35 nucleotides were defined to differentiate each of the HPV1a, 5, 6b, 8, 11, 16, 18 and 33. They were chosen using computer programs developed to compare simultaneously several 8000 bp long DNA sequences. Sequences common to all and to specific groups of the HPV DNA were also selected. Specificity of 32P-labelled probes for HPV6b, 11, 16, 18 and 33 was demonstrated and the sensitivity of the assays was evaluated by filter hybridization with viral clones and with DNA from cervical tumor biopsies. PMID- 2549407 TI - Oxidized apurinic/apyrimidinic sites formed in DNA by oxidative mutagens. AB - Treatment of DNA with any of several agents, including ionizing radiation, hydrogen peroxide, bleomycin, neocarzinostatin and the copper (I) chelate complex of 1,10-phenanthroline, produces apurinic/apyrimidinic (AP) sites containing oxidized deoxyribose moieties. These AP sites, which are formed by specific or nonspecific free-radical attack on deoxyribose, have been shown to involve oxidation of deoxyribose at the C-1', C-2' or C-4' position. Oxidized AP sites are generally more susceptible to chemical cleavage than normal AP sites, but are in some cases resistant to cleavage by repair AP endonucleases. Nearly all of the AP sites produced by neocarzinostatin, and a fraction of those produced by bleomycin, are accompanied by closely opposed breaks in the complementary strand. Sequence specificity data strongly implicate oxidized AP sites in neocarzinostatin-induced mutagenesis. The role of AP sites in mutagenesis by the other oxidative mutagens is less clear, although there is in some cases suggestive evidence for such a role. PMID- 2549408 TI - Characterization of hydroxyl free radical mediated damage to plasmid pBR322 DNA. AB - We have investigated hydroxyl free radical mediated damage to pBR322 DNA produced by ascorbate/iron and oxygen in a phosphate-buffered in vitro system. An observed lag phase in DNA nicking suggests a multi-target model of hydroxyl free radical attack on DNA. In the present report we further examine the model system and show that there is a "heat labile" component of the ascorbate/iron system which can be completely restored by the readdition of ascorbate. These observations have allowed us to rule out the possibility that intermediates build up in the reaction and act independently of ascorbate to increase the reaction rate. We have investigated the initial rate of OH production with two OH trapping agents, salicylate and deoxyguanosine, and find that the lag in DNA nicking is not due to a corresponding lag in the production of OH as assessed by formation of the products, dihydroxybenzoic acids and 8-hydroxydeoxyguanosine, respectively. We have found that the energy of activation for DNA supercoiled nicking is 13.9 kcal/mole and for OH trapping by salicylate is 21.1 kcal/nmole. These two activation energies are sufficiently different to suggest that the rate-limiting steps of these two reactions are different. Investigation of the rate of oxygen consumption during the ascorbate/iron-mediated DNA damage showed that oxygen was not a limiting component at any point in the reaction. The addition of catalase slowed down oxygen consumption by 31% and this data taken together with our previous observations on the model implicate hydrogen peroxide as a key intermediate in DNA damage caused by hydroxyl free radical. PMID- 2549409 TI - Comparison of gamma-radiation-induced accumulation of ataxia telangiectasia and control cells in G2 phase. AB - Recent reports from a number of laboratories have linked radiosensitivity in ataxia telangiectasia (A-T) to a large and prolonged block of some cells in G2 phase. Previous results from this laboratory, largely with one Epstein-Barr virus transformed A-T lymphoblastoid cell line, presented evidence for a dramatic increase in the number of cells in G2 phase over controls during a 24-h period post irradiation. We describe here a study of the effect of gamma-radiation on G2 phase delay in several A-T cell lines. Based on previous results with several cell lines 24 h post irradiation was selected as the optimum time to discriminate between G2 phase delay in control and A-T cells. All A-T homozygotes showed a significantly greater number of cells in G2 phase, 24 h post irradiation, than observed in controls. A more prolonged delay in G2 phase after irradiation was seen in different A-T cell types that included lymphoblastoid cells, fibroblasts and SV40-transformed fibroblasts. At the radiation dose used it was not possible to distinguish A-T heterozygotes from controls. PMID- 2549410 TI - Effect of combinations of antioxidants on phagocyte-induced sister-chromatid exchanges. AB - Combinations of oxygen radical scavengers and antioxidants significantly reduced the number of sister-chromatid exchanges in Chinese hamster ovary cells exposed to human phagocytes stimulated to generate oxygen radicals. When vitamin E was combined with these antioxidants, no increase in sister-chromatid exchanges was observed compared to controls. PMID- 2549411 TI - Increased chromosomal radiosensitivity in patients undergoing radioimmunoglobulin therapy. AB - Lymphocytes from individual patients undergoing radiolabeled immunoglobulin therapy have been examined both for chromosome aberrations expressed immediately upon explant, or for chromosome aberrations induced by a subsequent challenge of gamma-rays after PHA-stimulated proliferation. Despite interpatient variation, there is strong correlation between levels of chromosome aberrations observed in the initial mitosis after mitogenic stimulation and levels induced by a challenge dose of radiation in replicate cultures after several cell cycles of growth. These data indicated that even after proliferation, human lymphocytes retain a memory of in vivo exposure to ionizing radiation that can be observed by challenge with a clastogenic agent. This persistent hypersensitivity occurs at high frequency, suggesting that it may be related to initial steps in multistage carcinogenesis. PMID- 2549412 TI - Mutagenic activation of aflatoxin B1 by P-450 HFLa in human fetal livers. AB - The genotoxic and mutagenic activation of promutagens by human fetal livers was measured by the induction of umu gene expression in Salmonella typhimurium TA1535/pSk1002. Liver homogenates of human fetuses were the most active for the mutagenic activation of aflatoxin B1 (AFB1), followed by 2-amino-3 methylimidazo(4,5-f)quinoline (IQ), and to a lesser extent by 2-amino-6 methyldipyrido(1,2-a:3',2'-d)imidazole (Glu-P-1). The amounts of P-450 HFLa immunochemically determined in human fetal livers correlated highly with the induction of umu gene expression by AFB1 (r = 0.98, n = 5). P-450 HFLa catalyzed the mutagenic activation of AFB1 in a reconstituted system: cytochrome b5 markedly stimulated the activation. Anti-P-450 HFLa antibodies inhibited the mutagenic activation of AFB1 in a dose-dependent manner. These results strongly support the idea that P-450 HFLa is responsible for the mutagenic activation of AFB1 in human fetal livers. PMID- 2549413 TI - Abnormalities of the fast sodium current in myotonic dystrophy, recessive generalized myotonia, and adynamia episodica. AB - Myoballs, i.e., spherical muscle cell regenerates, were cultured from the biopsied muscles of three patients with myotonic dystrophy, three patients with recessive generalized myotonia, and a patient with adynamia episodica. The membrane of these myoballs was voltage-clamped in the whole-cell mode for the recording of sodium currents (at 11, 24, and 37 degrees C). The voltage dependence of the steady-state activation and inactivation curves showed only minor abnormalities in all cases. The time constants of activation (tau m) and inactivation (tau h), when studied at the three temperatures, showed a characteristic pattern of abnormalities. In myotonic dystrophy, both tau m and tau h were larger than control; in recessive generalized myotonia and adynamia episodica both tau m and tau h were smaller than control. In the latter diseases, these time constants also showed a smaller than normal decrease with membrane depolarization. The changes seen for recessive generalized myotonia and adynamia episodica would favour the occurrence of myotonia, the opposite results for myotonic dystrophy would oppose myotonia. PMID- 2549414 TI - Muscle glycerol kinase in Duchenne dystrophy and glycerol kinase deficiency. AB - The complex glycerol kinase deficiency (GKD) syndrome is an X-linked recessive genetic disorder. The syndrome often includes a myopathy that is similar histologically to Duchenne muscular dystrophy (DMD). The glycerol kinase (GK) locus is in the Xp21 region in the midportion of the short arm of the X chromosome and is in close proximity to the DMD locus. We have investigated GK activity and subcellular distribution of muscle GK in DMD patients and in a patient with the complex GKD syndrome presenting with myopathy. We found no abnormality of muscle GK specific activity or subcellular distribution in DMD. In the patient with the complex GKD syndrome the specific activity and kinetics of muscle GK were normal, but the subcellular distribution of muscle GK was altered. Liver GK had less than 10% of normal activity and showed markedly altered kinetics. These findings indicate that there is no abnormality of muscle GK activity in DMD muscle. Furthermore, the normal GK activity in an individual with the complex GKD syndrome suggests that muscle and liver GK are genetically distinct. These findings support the concept that the complex GKD syndrome results from small deletions that affect closely linked but separate loci for DMD, GK and adrenal hypoplasia. PMID- 2549415 TI - Therapeutic strategies in promoting peripheral nerve regeneration. AB - Currently, regeneration chambers, adrenocorticotropic hormone (ACTH) and related peptides, and gangliosides appear to be the most promising therapies in the promotion of peripheral nerve regeneration, growth, and repair. Regeneration chambers enhance rat sciatic nerve regeneration in vivo after transection by providing a structurally organized and protected preformed space within which nerve fibers are exposed to macromolecular compounds which direct and enhance nerve growth. ACTH and related peptides, independent of their corticotropic activities, increase the availability of structural proteins to the axon terminal in rats subjected to nerve crush injuries and demonstrate inotropic effects in adrenalectomized and/or hypophysectomized rats. Exogenously administered gangliosides promote neuronal sprouting, regeneration, and reinnervation in experimental situations and have undergone clinical testing in acute and chronic peripheral nerve disorders. At the current dosage levels and schedules, the clinical results of ganglioside therapy have been mixed. The success of the experimental studies supports further clinical testing of these therapies in peripheral nerve disorders. PMID- 2549416 TI - Biochemical characteristics of free and junctional sarcoplasmic reticulum and of transverse tubules in human skeletal muscle. AB - The microsomal fraction of normal human skeletal muscle was subfractionated by isopycnic sucrose-density centrifugation, using the procedure originally described by Saito et al. for rabbit fast muscle, and specific markers of the junctional face membrane of terminal cisternae (TC) (ryanodine receptor, high molecular-weight feet proteins and membrane-associated calcium-binding protein calsequestrin), of the sarcoplasmic reticulum (SR) Ca-pump membrane (chicken antibody to rabbit Ca-ATPase), and of transverse tubules (TT) (dihydropiridine receptor, membrane cholesterol), respectively. The results show that isolated TC from human skeletal muscle share extensive morphological characteristics, protein composition, as well as Ca-release properties with rabbit TC, as tested with an inhibitor (Ruthenium red) and an activator (doxorubicin) of SR Ca-release. The Ca pump membrane of human muscle SR, in distinction to rabbit fast muscle SR, showed a relatively low specific activity of the Ca-ATPase, as expected from the mixed fiber composition of human muscles, but shared the presence of minor protein components, such as a Con A binding protein of about 57 kDa and blue-staining peptides in the 170-120 kDa range of molecular weights. Human muscle TT, as isolated from the same sucrose gradient, demonstrated a high affinity (3H) dihydropiridine binding activity in the range of previously reported values for purified TT from rabbit skeletal muscle. PMID- 2549417 TI - Sequences similar to genes for two mitochondrial proteins and portions of ribosomal RNA in tandemly arrayed 6-kilobase-pair DNA of a malarial parasite. AB - Erythrocytic stages of mammalian malarial parasites contain acristate mitochondria whose functions are not well understood. Moreover, little is known about the genome of these organelles. We have previously reported that all species of malarial parasites examined contain highly conserved, tandemly arrayed DNA with a unit length of about 6.0 kb that is transcribed into discrete RNA molecules in erythrocytic stages. We now report the complete DNA sequence of the 5984-bp repeating unit of Plasmodium yoelii, a rodent parasite. Two slightly overlapping regions transcribed into large RNA molecules were found to have significant DNA and protein sequence similarity with mitochondrion-coded proteins, cytochrome c oxidase subunit I and cytochrome b. Significant sequence similarity with other mitochondrial protein genes could not be detected. Ribosomal RNA (rRNA)-like genes were not detected in this sequence either. However, two regions, 82 and 50 nucleotides long, specified by different strands, were found to have extensive similarity with the highly conserved central loop of the peptidyl transferase domain of the large rRNA of Escherichia coli, mitochondria, and chloroplasts. Compensatory nucleotide substitutions were present in these regions, so that the predicted secondary structure was not affected. Functional utilization of these regions, if it exists, could argue for a trans-associative origin of rRNA. In organization, size and sequence, the tandem arrays of 6.0 kb malarial DNA appear to be a very unusual form of mitochondrial DNA. PMID- 2549419 TI - Calcium-antagonist receptors in cardiomyopathy. PMID- 2549418 TI - Maintenance of cytoplasmic pH and proton motive force in promastigotes of Leishmania donovani. AB - Three methods were used to measure intracellular pH (pHi) of Leishmania donovani promastigotes: (a) measurement of the fluorescence of the pH indicator 2',7'-bis (carboxyethyl)-5,6-carboxyfluorescein; (b) pH null point assays; and (c) determination of the distribution across the promastigote plasma membrane of the fluorescent amine acridine orange and of the weak acid 5,5-dimethyl-2,4 oxazolidinedione. The three methods gave similar results and showed that promastigotes of L. donovani maintain pHi at a narrow range of 6.4-6.7, throughout an extracellular pH (pHo) range of 5.5-7.4. L-Proline transport in L. donovani promastigotes, which is known to be coupled to proton translocation, was used to estimate the proton electrochemical gradient across parasite plasma membrane. While proline uptake is optimal at pHo 7.5, an outward-directed concentration gradient is obtained at steady state throughout a pHo range of 5-8. The calculated electrochemical gradient of proline across the parasite plasma membrane at steady state is 90-100 mV within a pHo range of 5-8, suggesting an almost constant proton electrochemical gradient at this pHo range. Taken together, the results show that the parasites regulate both pHi and the size of the chemiosmotic energy required to drive active transport of nutrients. PMID- 2549420 TI - Fast-food fare. Consumer guidelines. PMID- 2549421 TI - Itraconazole vs amphotericin B: in vitro comparative evaluation of the minimal inhibitory concentration (MIC) against clinically isolated yeasts. AB - Itraconazole is a triazole compound which, following several clinical trials, has begun to be used for therapy of mycotic infections. This new drug, with a broad spectrum antifungal activity, can be orally administered. The Authors studied the in vitro susceptibility to amphotericin B and itraconazole of the following clinical isolates of pathogenic yeasts: 100 Candida albicans, 20 C. tropicalis, 20 C. parapsilosis, 8 C. guilliermondii, 6 C. pseudotropicalis, 24 Torulopsis glabrata and 16 Cryptococcus neoformans. Serial two-fold dilution, from 100 micrograms/ml to 0.04 micrograms/ml, of each drug were prepared in Yeast Nitrogen Base + Glucose 5%, after dissolving the itraconazole in dimethylsulfoxide (DMSO) and amphotericin B in 5% glucose solution. Amphotericin B (MIC90: 3.12 micrograms/ml) was found to have an average in vitro MIC six-fold lower than itraconazole (MIC90: 25 micrograms/ml). Thus, even though itraconazole is active, amphotericin B remains one of the most effective of the antifungal drugs. PMID- 2549422 TI - Effects of lipolytic and antilipolytic agents on glycerol and free fatty acid release from isolated adipocytes of normal and diabetic rats. AB - Isolated adipocytes from severely diabetic rats exhibited hypersensitivity to epinephrine at low concentrations (0.05-0.1 microM) on lipolysis, compared with isolated adipocytes from normal and mildly diabetic rats. Hypersensitivity to dibutyryl cyclic AMP and theophylline at concentrations from 0.05 to 0.50 mM was not observed in adipocytes of severely diabetic rats. Insulin could not exert an inhibitory effect on epinephrine-induced lipolysis in adipocytes of severely diabetic rats. In isolated adipocytes from normal rats, hyperosmolarity due to the combination of 50 mM glucose and 100 mM sodium chloride only had an inhibitory effect on 0.25 microM epinephrine-induced lipolysis. Ten mM beta hydroxybutyrate did not inhibit lipolysis caused by epinephrine although any lipolysis stimulated by epinephrine, dibutyryl cyclic AMP and theophylline was inhibited by insulin. Our present findings may partly explain insulin resistance in the severely diabetic state and the pathogenesis of the absence of ketosis in hyperglycemic hyperosmolar conditions. PMID- 2549423 TI - An essential role for postsynaptic calmodulin and protein kinase activity in long term potentiation. AB - The phenomenon of long-term potentiation (LTP), a long lasting increase in the strength of synaptic transmission which is due to brief, repetitive activation of excitatory afferent fibres, is one of the most striking examples of synaptic plasticity in the mammalian brain. In the CA1 region of the hippocampus, the induction of LTP requires activation of NMDA (N-methyl-D-aspartate) receptors by synaptically released glutamate with concomitant postsynaptic membrane depolarization. This relieves the voltage-dependent magnesium block of the NMDA receptor ion channel, allowing calcium to flow into the dendritic spine. Although calcium has been shown to be a necessary trigger for LTP (refs 11, 12), little is known about the immediate biochemical processes that are activated by calcium and are responsible for LTP. The most attractive candidates have been calcium/calmodulin-dependent protein kinase II (CaM-KII) (refs 13-16), protein kinase C (refs 17-19), and the calcium-dependent protease, calpain. Extracellular application of protein kinase inhibitors to the hippocampal slice preparation blocks the induction of LTP (refs 21-23) but it is unclear whether this is due to a pre- and/or postsynaptic action. We have found that intracellular injection into CA1 pyramidal cells of the protein kinase inhibitor H-7, or of the calmodulin antagonist calmidazolium, blocks LTP. Furthermore, LTP is blocked by the injection of synthetic peptides that are potent calmodulin antagonists and inhibit CaM-KII auto- and substrate phosphorylation. These findings demonstrate that in the postsynaptic cell both activation of calmodulin and kinase activity are required for the generation of LTP, and focus further attention on the potential role of CaM-KII in LTP. PMID- 2549424 TI - Dual regulatory role for thyroid-hormone receptors allows control of retinoic acid receptor activity. AB - Both thyroid hormone (T3) and retinoic acid signal essential steps in development, differentiation and morphogenesis. Specific nuclear receptors for these ligands have recently been cloned. Previously we have noted a close homology between the DNA-binding domains of the epsilon-retinoic acid receptor (RAR-epsilon, also designated RAR-beta), the thyroid hormone receptors and the oestrogen receptor. We have now found that RAR-epsilon is very efficient at inducing transcription from two distinct thyroid-hormone responsive elements (TREs). Transcription induced by ligand-activated RAR-epsilon from a TRE can, however, be repressed by thyroid-hormone receptor in the absence of its ligand. Conversely, in the presence of its ligand, thyroid-hormone receptor will activate transcription from a TRE irrespective of the presence of unbound RAR. The use of hybrid receptors has shown that the DNA-binding domain of RAR is the essential target for inhibition by thyroid-hormone receptors. These data, together with in vitro DNA-binding studies, suggest that thyroid-hormone receptors may have dual regulatory roles: in the presence of hormone they function as TRE-specific transcriptional activators; in the absence of hormone, however, they can function as TRE-specific repressors. PMID- 2549425 TI - Similarities between prokaryotic and eukaryotic cyclic AMP-responsive promoter elements. AB - Organisms as diverse as bacteria and man contain genes that show transcriptional induction when the intracellular concentration of cAMP is increased. This regulated transcriptional response is mediated through specific promoter elements located, in general, upstream from the transcription start site. In Escherichia coli the element responsible for cAMP-mediated transcriptional induction is the binding site for the cAMP-receptor protein (CAP). In mammalian cells the cAMP regulatory element is composed of one or more binding sites for various transcription factors. In many instances the cAMP regulatory element contains binding sites for a family of proteins referred to as ATF. Here we provide evidence that some prokaryotic and mammalian cAMP-response elements are functionally related. First, we show that mammalian ATF binds specifically to some E. coli CAP sites, and conversely E. coli CAP binds specifically to some mammalian ATF sites. Second, we demonstrate that an E. coli CAP binding site can confer cAMP-inducibility onto a mammalian gene when assayed in transfected mammalian cells. PMID- 2549426 TI - GTPase inhibiting mutations activate the alpha chain of Gs and stimulate adenylyl cyclase in human pituitary tumours. AB - A subset of growth hormone-secreting human pituitary tumours carries somatic mutations that inhibit GTPase activity of a G protein alpha chain, alpha(s). The resulting activation of adenylyl cyclase bypasses the cells' normal requirement for trophic hormone. Amino acids substituted in the putative gsp oncogene identify a domain of G protein alpha-chains required for intrinsic ability to hydrolyse GTP. This domain may serve as a built-in counter-part of the separate GTPase-activating proteins required for GTP hydrolysis by small GTP-binding proteins such as p21ras. PMID- 2549427 TI - Calcitonin gene-related peptide regulates calcium current in heart muscle. AB - The influx of Ca2+ due to the transmembrane calcium current, ICa, has a fundamental role in cardiac pacemaker activity, in the action potential plateau and in excitation-contraction coupling. Both sympathetic and parasympathetic neurotransmitters can modulate ICa. Recent studies indicate that in both the cardiovascular and the central nervous systems, nerve varicosities exist that contain a novel non-adrenergic, non-cholinergic peptide--calcitonin gene-related peptide (CGRP). Although CGRP is known to exert strong positive inotropic and chronotropic effects, as well as to cause vasodilation, very little is known about the ionic mechanisms of these effects. Here we report that CGRP dramatically increases ICa in single heart cells. Although this CGRP-induced increase in ICa resembles the effect of beta-adrenergic agonists, our results demonstrate some significant differences between the effects of CGRP and these agonists: (1) the increase due to CGRP cannot be blocked by beta-adrenergic antagonists; (2) the CGRP-induced effect is transient; and, (3) CGRP can inhibit isoproterenol-stimulated ICa. Our results provide the first electrophysiological evidence that CGRP can significantly modulate ICa in the heart, and suggest a new additional mechanism for the neurogenic control of cardiac function. PMID- 2549428 TI - Augmentation of cardiac calcium current by flash photolysis of intracellular caged-Ca2+ molecules. AB - The entry of calcium ions into cells through voltage-activated Ca2+ channels in the plasma membrane triggers many important cellular processes. The activity of these channels is regulated by several hormones and neurotransmitters, as well as intracellular messengers such as Ca2+ itself (for examples, see refs 1-9). In cardiac muscle, myoplasmic Ca2+ has been proposed to potentiate Ca2+ influx, although a direct effect of Ca2+ on these channels has not yet been demonstrated. Photosensitive 'caged-Ca2+' molecules such as nitr-5, however, provide powerful tools for investigating possible regulatory roles of Ca2+ on the functioning of Ca2+ channels. Because its affinity for Ca2+ is reduced by irradiation, nitr-5 can be loaded into cells and induced to release Ca2+ with a flash of light. By using this technique we found that the elevation of intracellular Ca2+ concentration directly augmented Ca2+-channel currents in isolated cardiac muscle cells from both frog and guinea pig. The time course of the current potentiation was similar to that seen with beta-adrenergic stimulation. Thus Ca2+ may work through a similar pathway, involving phosphorylation of a regulatory Ca2+-channel protein. This mechanism is probably important for the accumulation of Ca2+ and the amplification of the contractile response in cardiac muscle, and may have a role in other excitable cells. PMID- 2549429 TI - Effects of pertussis toxin on opioid regulation of catecholamine release from rat and guinea pig brain slices. AB - Opioid agonists selective for mu-, delta-, and kappa-receptors are all capable of regulating the stimulated release of noradrenaline from three terminal fields (cortex, hippocampus, and cerebellum) of the noradrenergic projections from locus coeruleus in the guinea pig brain. Intracerebroventricular injections of pertussis toxin abolished the ability of a mu-selective agonist and of a delta selective agonist to inhibit stimulated noradrenaline release, but left unaffected the concentration-related inhibition of NE release by a kappa agonist. Thus, mu- and delta-receptors have been shown to be coupled to their effector system in these noradrenergic neurons via guanyl nucleotide binding proteins (G proteins) which are sensitive to pertussis toxin, while kappa-receptors in the same neurons appear to be coupled through a different mechanism which is significantly less sensitive to pertussis toxin. In contrast to opioid receptor regulation of noradrenaline release in guinea pig hippocampus, mu-, but not delta or kappa-agonists are capable of regulation of stimulated noradrenaline release from rat hippocampus and cortex, and kappa-, but not mu- or delta-agonists are capable of inhibiting the stimulated release of dopamine from rat striatum and cortex. Pertussis toxin injections significantly attenuated mu-agonist inhibition of noradrenaline release, but had no effect on the ability of a kappa-selective agonist to regulated dopamine release, confirming the insensitivity of the kappa receptor-effector coupling system to pertussis toxin. PMID- 2549430 TI - Relation of positive inotropic and chronotropic effects of pimobendan, UD-CG 212 Cl, milrinone and other phosphodiesterase inhibitors to phosphodiesterase III inhibition in guinea-pig heart. AB - (1) This study was performed to elucidate the relation between positive inotropy and phosphodiesterase inhibition in the heart. Therefore, the influence on the activity of guinea-pig cardiac phosphodiesterase (PDE) I-III separated by DEAE cellulose anion exchange chromatography was investigated for the new cardiotonic agents pimobendan, its metabolite UD-CG 212 Cl and milrinone. These effects were compared with those of various other PDE inhibitors such as IBMX, zaprinast, rolipram and AR-L 57 Cl. A selectivity factor (SF, mean of the IC50 values for PDE I and II inhibition divided by the IC50 for PDE III) was calculated for each drug. The greater this value the more selective was the PDE III inhibition. (2) UD-CG 212 Cl was the most potent (IC50 = 0.19 mumol/l) and most selective inhibitor of PDE III with a SF of 869. Also selective PDE III inhibitors were pimobendan (SF = 50.5) and milrinone (SF = 70.0) with slightly smaller potencies (IC50 = 2.40 and 1.52 mumol/l, respectively). Zaprinast and rolipram preferentially inhibited PDE I and II, respectively. IBMX and AR-L 57 Cl inhibited PDE I-III unselectively with similar potencies for all isoenzymes. (3) The PDE inhibitory effects of all substances were compared with their influence on force of contraction (electrically driven papillary muscles) and on frequency of beating (spontaneously beating right auricles) in guinea-pig hearts, thus in preparations of the same species. UD-CG 212 Cl and pimobendan resembled each other in their maximal positive inotropic effects with potencies (EC50) of 1.8 mumol/l and 6.0 mumol/l, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549433 TI - [Effect of tractotomy on the reflex activity of the lumbar segment of the spinal cord severed after sectioning of the sciatic nerve in the rat]. AB - Electrical responses of the spinal cord roots of rats evoked by the preliminary cutting of nerve were investigated in 1, 3, 7 and 14 days after tractotomy. Mean amplitude of the monosynaptic reflex discharges in ventral roots on the side of nerve section is substantially higher 7 and 14 days after the tractotomy, than that of these responses 1 and 3 days after it. PMID- 2549432 TI - In vitro inhibition of human polymorphonuclear cell function by cloricromene. AB - Cloricromene, a coumarin derivative with antiaggregating and vasodilating properties, was tested in vitro on polymorphonuclear cell (PMN) adhesion to the endothelium, superoxide anion generation and chemotaxis. PMN adhesion was measured using cultured human umbilical vein endothelial cells (EC) either untreated or previously activated with interleukin-1 (IL-1). Cloricromene (5-50 microM) induced dose-related inhibition of PMN adhesion to untreated and IL-1 treated EC. Cloricromene also inhibited PMN superoxide generation induced by the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA) or by N-formyl methionyl-leucyl-phenylalanine (FMLP). PMN and monocyte chemotaxis was evaluated by a modification of the Boyden chamber technique. Cloricromene inhibited both types of cell motility induced by FMLP in a concentration-dependent fashion. The major cloricromene metabolite (cloricromene acid) had no effect on any of the biological parameters studied up to a concentration of 500 microM. HPLC measurement showed that cloricromene accumulated in PMN within a few minutes and levels of the drug were still high after 60 min. In contrast its acid metabolite was not taken up in a significant amount during incubation periods up to 60 min. We conclude that cloricromene inhibits a series of PMN activities in vitro. This effect might be of pharmacological interest in view of the role of PMN activation in different pathophysiological conditions. PMID- 2549431 TI - Action of anticonvulsants on hippocampal slices in Mg-free medium. AB - The effects of six prototype anticonvulsant drugs were investigated on epileptiform field potential discharges evoked in hippocampal slices of rats by removing magnesium ions from the perfusion fluid in order to reveal a possible interaction with N-methyl-D-aspartate (NMDA) receptor activation. All drugs reduced the multiple discharges with the following order of potency: midazolam greater than carbamazepine = phenytoin = phenobarbital greater than ethosuximide = valproate. They had a stronger depressant effect on the later population spikes but none of them abolished the epileptiform discharge. These effects can be explained by known mechanisms of action of the anticonvulsants tested and lend no support for a specific interaction with NMDA receptors. PMID- 2549434 TI - [Changes in the intracellular calcium concentration and transmembrane currents, evoked by iontophoretic injections of cyclic AMP, in Helix pomatia neurons]. AB - Transmembrane currents and intracellular concentration of free Ca2+ were measured in voltage-clamped isolated neurons of Helix pomatia following the injection of cAMP. In most neurons in the range of membrane potentials from -40 to -100 mV cAMP injection induced both inward current and a long-lasting increase in [Ca2+]in. In the Ca-free external medium and after addition of EGTA (a Ca chelator) to it, the cAMP-induced inward current and [Ca2+]in increase remained unchanged. In most cases in Na-free external solution the cAMP-induced inward current markedly decreased, whereas [Ca2+]in changes remained as it were. Cd2+ (2 mM) did not affect the cAMP-induced current and [Ca2+]in increase. Both procaine++ and ryanodine (inhibitors of Ca release from intracellular stores) did not change the cAMP-induced effects. La3+ (1 mM) blocked both the inward current and an increase of [Ca2+]in. Obtained data confirm the hypothesis of cAMP mediated Ca release from the intracellular stores. PMID- 2549435 TI - A patient with a metastasizing jejunal carcinoma 17 years after colectomy for familial polyposis coli. AB - A patient is described who died of a metastasizing jejunal carcinoma 17 yr after colectomy for familial polyposis coli. A survey of the literature concerning gastric and small intestine polyps, malignant degeneration and implications for further management is presented. PMID- 2549436 TI - Circadian variation of beta-adrenoceptor binding sites in the pineal gland of the Syrian hamster and prevention of the nocturnal reduction by light exposure or propranolol treatment. AB - Radioreceptor assays with (-) [125I]-iodopindolol (IPIN) were used to describe circadian variations of beta-adrenoceptors in the pineal gland of male Syrian hamsters (Mesocricetus auratus). Single-point experiments (200 pM IPIN, 8-9 pineals pooled per time point) showed a significant circadian variation (p less than 0.01) with maximal values (28.9 +/- 1.8 fmol/mg protein, means +/- SEM, n = 7) between 07:00 and 02:30 h, and minimal values at 04:00 (7.2 +/- 2.9 fmol/mg protein, n = 3), 8 h after darkness onset. Either exposure of animals to light at night or treating dark exposed hamsters with a beta-adrenergic receptor blocker, propranolol, prevented the nocturnal drop in the number of beta-adrenoceptors. Scatchard analysis of saturation isotherms at 02:30, 04:00 and 08:00 h (30-600 pM IPIN, one saturation experiment with 25 pineals pooled per time point) confirmed the circadian variation. PMID- 2549437 TI - Neurotensin activates tuberoinfundibular dopamine neurons and increases serum corticosterone concentrations in the rat. AB - The activity of tuberoinfundibular dopamine neurons, as estimated from the amount of dihydroxyphenylalamine (DOPA) formed in the median eminence after the inhibition of DOPA decarboxylase and the concentration of dihydroxyphenylacetic acid (DOPAC) in this brain region, was significantly increased 1-8 h following the intracerebroventricular (i.c.v.) administration of neurotensin (20 micrograms). Neurotensin (5 and 20 micrograms i.c.v.) also significantly increased DOPAC concentrations in the n. accumbens but had no effect in the striatum. Serum concentrations of corticosterone in rats treated with neurotensin (1-20 micrograms i.c.v.) were 5-7 times those in vehicle-treated animals. [D Trp11]-neurotensin (0.5 micrograms i.v.c.) also significantly increased DOPAC concentrations in the median eminence and serum corticosterone concentrations. It is concluded that neurotensin acutely increases the activity of tuberoinfundibular and mesolimbic dopamine neurons and the secretion of ACTH. PMID- 2549438 TI - Secretion of melanocyte-stimulating hormone and adrenocorticotropin from transplanted pituitary pars intermedia in stressed and nonstressed rats. AB - Rats bearing kidney grafts of the pituitary pars intermedia were divided into three groups: unstressed, acutely stressed, and chronically stressed. Corresponding sham-operated rats were used for comparisons. Twenty days after grafting, the rats were sacrificed and alpha-melanocyte-stimulating hormone (alpha-MSH), adrenocorticotropin (ACTH), and corticosterone were estimated in plasma. The adrenal/body weight ratio and DNA content of the glands were also investigated. The following results were obtained: MSH was found not to be increased in unstressed rats, but it was in grafted animals subjected to acute and chronic swimming stress. ACTH and corticosterone rose in all three groups. Adrenal/body weight ratio and DNA content increased only in grafted chronically stressed rats. Moreover, plasma corticosterone was found higher in grafted hypophysectomized rats than in non-grafted hypophysectomized animals. Administration of ergocryptine to nonstressed grafted rats induced a decrease in the blood content of ACTH and MSH, indicating that the grafts were the source of a part of the circulating ACTH. On the other hand, the fall in MSH levels could show the effect of the drug upon the pars intermedia. Comparison of the ratios of both hormones released in incubations showed that grafts secreted more ACTH than MSH; on the other hand, when intact neurointermediate lobes were incubated, MSH predominated over ACTH. For the first time it is demonstrated that the pars intermedia can secrete ACTH in vivo. Nevertheless, the ability to secrete this hormone is not a property of normal intact pars intermedia, but it manifests in the transplantations probably due to the overactivity of light cells induced by chronic stoppage of dopaminergic inhibition. PMID- 2549439 TI - Dose response effects of pulsatile GnRH administration on restoration of pituitary GnRH receptors and pulsatile LH secretion during lactation. AB - Ovariectomized (OVX) rats suckling 8 pups have a complete suppression of pulsatile LH secretion and a decrease in pituitary GnRH receptor (GnRH-R) content. Removing the suckling stimulus for 24 h results in a sharp increase in GnRH-R and a restoration of pulsatile LH secretion. These findings suggest that the suckling stimulus induces a suppression of GnRH secretion, and removal of the suckling stimulus permits the restoration of GnRH secretion. Indeed, if GnRH antiserum is injected at the time of pup removal, the restoration of pituitary GnRH-R and LH secretion is prevented. The present studies were designed to test our hypothesis that the deficits in pituitary gonadotroph function observed during lactation are due to suckling-induced suppression of GnRH. Exogenous GnRH was administered in a pulsatile regimen to OVX lactating rats on days 10 and 11 postpartum, and the effects on pituitary GnRH-R levels, pituitary sensitivity to GnRH, and pulsatile LH secretion were assessed. GnRH doses of 0, 0.5, 2.0 or 5.0 ng/pulse were administered every 50 min for 24 h beginning on day 10. Administration of 0.5 ng GnRH/pulse for 24 h increased GnRH-R from 35 +/- 3 to 63 +/- 8 fmol/pituitary. There was a clear GnRH dose-related upregulation of GnRH-R to approach nonsuckling levels (140-160 fmol/pituitary) with the 5 ng GnRH dose. At the beginning of GnRH administration, the pituitary was very unresponsive to GnRH. Consistent LH pulses were only observed with 5 ng GnRH/pulse.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549440 TI - Effect of corticotropin-releasing factor on the release and synthesis of prolactin. AB - Corticotropin-releasing factor (CRF) has been characterized on the basis of its intrinsic activity to release corticotropin from cultured rat anterior pituitary cells. Injected in intact rats, CRF increases adrenocorticotropic hormone (ACTH) release. Endogenous CRF-like immunoreactivity was detected in the cytoplasm and nucleus of corticotrophs. Using an antirat CRF serum, a similar location of CRF like immunoreactivity was observed in lactotrophs: cytoplasmic matrix, secretory granules, nucleus and, to a lesser degree, the plasma membrane level were stained. One injection of CRF increased the plasma ACTH concentration 4-fold after 15 min, while plasma prolactin (PRL) increased 2.7-fold 5 min after injection. In vitro, incubation of female pituitary cells with rat CRF (10(-10) 10(-8) M) had no significant effect on PRL secretion. In contrast, after 4 days of in vitro pretreatment with 17 beta-estradiol (10(-9) M), rat CRF stimulated PRL secretion by 42%. In situ hybridization of whole pituitary slices showed that rat CRF injection significantly increased the labeling of corticotrophs using an ACTH-cDNA probe, but had no significant effect on the labeling of lactotrophs using a PRL riboprobe. These results indicate that CRF is a factor which can modulate PRL release but not the synthesis of PRL. PMID- 2549441 TI - Corticotropin-releasing factor secretion increases in rat hypophysial portal blood during insulin-induced hypoglycemia. AB - To study the possible involvement of hypothalamic corticotropin-releasing factor (CRF) in the stimulation of adrenocorticotropic hormone (ACTH) release caused by insulin-induced hypoglycemia (IIH), we measured CRF secretion in hypophysial portal blood (HPB) in rats anesthetized with sodium thiopental after injection of insulin. Before treatment, systemic ACTH levels (952 +/- SE 143 pg/ml; n = 12) were well above normal values, probably reflecting the anesthetic and surgical stress consecutive to the preparation for portal blood collection. Insulin injection induced a significant increase of ACTH release within 15 min (1,588 +/- 168 vs. 741 +/- 144 pg/ml; n = 6, in vehicle-injected rats) which lasted for at least 1 h. CRF levels in HPB were 857 +/- SE 168 pg/ml (n = 13) during the first hour pretreatment collection. Vehicle injection did not modify CRF secretion (759 +/- 142 pg/ml; n = 6). Insulin injection provoked a significant increase in CRF release (1,449 +/- 257 pg/ml; n = 7). These data suggest that an increased hypothalamic CRF secretion is responsible for the stimulation of pituitary ACTH release following IIH. The possible involvement of central neuromediators in the IIH-induced CRF production is discussed. PMID- 2549442 TI - Stereotactic biopsy and resection of thalamic astrocytomas. AB - In this study of 72 patients who had histologically verified thalamic astrocytomas, 44 patients underwent stereotactic serial biopsy, 22 underwent stereotactic resection of the neoplasm, and an additional 6 patients underwent stereotactic biopsy followed by stereotactic resection of the tumor at a later date. Of the 50 patients who underwent stereotactic biopsy, 3 were neurologically worse after the procedure (morbidity, 6%), and 3 additional patients with Grade 4 astrocytomas who preoperatively were rapidly deteriorating neurologically, died within 30 days of the procedure. Of the 28 patients who underwent stereotactic resection, 14 were neurologically improved after the procedure, 10 were unchanged, and 4 were worse. One additional patient died 10 days postoperatively. Thirty-four patients had Grade 4 astrocytomas: 27 underwent stereotactic biopsies. The mean survival after biopsy and irradiation for patients with Grade 4 astrocytomas was 21.4 weeks. The mean survival was 62 weeks in 7 patients with Grade 4 astrocytomas who underwent stereotactic resection and radiation therapy. The mean survival time after biopsy and radiation therapy for patients who had Grade 3 and Grade 2 lesions was 54.4 weeks and 91 weeks, respectively. Twenty three patients had pilocytic astrocytomas; 8 underwent stereotactic biopsies, and 19 underwent stereotactic resection of the tumor (4 of these underwent biopsy prior to resection). There was no neurological morbidity, but one patient died after resection. Many of those who underwent resection were deteriorating due to an enlarging tumor mass or recurring cyst, and had undergone more conservative therapies such as biopsy and radiation. Even though stereotactic biopsy is appropriate in many patients harboring thalamic astrocytomas, selected patients with significant mass effect from solid tumor or recurring cyst can benefit from stereotactic resection. PMID- 2549443 TI - Reconstruction of cranial defects with porous hydroxylapatite blocks. AB - Repair of cranial defects for brain protection and aesthetics is currently a surgical problem for which there is no completely satisfactory solution. Material used for repair of cranial defects should ideally be incorporated by the body and provided a blood supply. No substance to date, including autologous bone, consistently achieves this. Hydroxylapatite is a polycrystalline, nonresorptive, biocompatible ceramic that allows osseous tissue ingrowth and ultimate transformation into vascularized bone. The authors have used granular hydroxylapatite successfully for reconstructing calvarial defects, but have experienced problems with migration of granules, prolonged mobility, and, in one patient, resorption of an underlying supporting bone graft. The use of hydroxylapatite in block form precludes most of these problems. Porous block or strips are easily contoured and can be fixed at the margin of the calvarial defect, providing immediate stability. Ingrowth of osteoblastic and fibrous tissue provides added strength and stability. This paper describes our experience with porous hydroxylapatite blocks for reconstructive cranioplasty. PMID- 2549444 TI - Simultaneous intracellular and focal extracellular recording of junction potentials and currents, and the time course of quantal transmitter action in rodent vas deferens. AB - Simultaneous recordings were made of spontaneous excitatory junction potentials and the underlying spontaneous excitatory junction currents in guinea-pig and mouse vas deferens using adjacent intracellular and focal extracellular electrodes. Concurrent spontaneous excitatory junction potentials and spontaneous excitatory junction currents were observed in a small proportion of smooth muscle cells penetrated intracellularly within 50-200 microns of the extracellular electrode. These simultaneous events had identical variations in time course, indicating that they were caused by the same transmitter release event. Their amplitudes were not related. Concurrent spontaneous excitatory junction potentials and currents had identical durations, rise times and time constants of decay, showing that the spontaneous excitatory junction potential reflects the time course of quantal transmitter action. In contrast, spontaneous "discrete events" obtained by differentiating the rising phases of spontaneous excitatory junction potentials with respect to time were brief compared with the underlying currents. Excitatory junction potentials evoked by electrical stimulation of the hypogastric nerve were prolonged compared to the underlying excitatory junction currents. The peaks in the first time differential of individual excitatory junction potentials (evoked discrete events) were brief compared to corresponding excitatory junction currents. It is concluded that at the neuroeffector junction of the rodent vas deferens the membrane potential response to a quantum of spontaneously released transmitter is a good estimate of the duration of transmitter action, in accordance with some of the predictions for three dimensional syncytial tissues. The first time differential of the membrane potential, the "discrete event", does not reflect the time course of spontaneous or evoked quantal transmitter action in these syncytial tissues. PMID- 2549445 TI - Nerve growth factor can influence growth of cortex cerebri and hippocampus: evidence from intraocular grafts. AB - The effects of nerve growth factor and antiserum against nerve growth factor on cortical cholinergic projection areas in the central nervous system and cerebellum were evaluated using intraocular grafts of cortex cerebri, hippocampus and cerebellum in rat hosts receiving injections into the anterior chamber of the eye of nerve growth factor (at transplantation, 5 and 10 days after transplantation) or antiserum to nerve growth factor (every 5 days). The controls received cytochrome c or preimmune serum. Growth of grafts was followed by repeated observations directly through the cornea of the host using a stereomicroscope. Nerve growth factor-treated grafts of cortex cerebri and hippocampus grew significantly smaller as compared to the corresponding control grafts. In one experiment, growth of cytochrome c and saline-treated cortex cerebri was compared and no difference in growth was found. Growth of nerve growth factor-treated cerebellar grafts did not differ significantly from growth of cytochrome c-treated grafts. Morphological analysis using Nissl-staining, antibodies to glial acidic fibrillary protein to evaluate the degree of gliosis and antiserum to neurofilament as a neuronal marker did not reveal any marked differences between nerve growth factor- and cytochrome c-treated grafts. Cortical grafts receiving anti-nerve growth factor antiserum by injection or by immunizing host rats against nerve growth factor showed similar growth to the controls. Similarly, grafts of fetal hippocampus to rats immunized with nerve growth factor were not significantly different from grafts to host rats immunized with cytochrome c. We conclude that exogenous nerve growth factor affects the development of grafted cortex cerebri and hippocampus. The fact that these cortical areas stop growing earlier in the presence of nerve growth factor without the grafts showing evidence of disturbed glial or neuronal populations compared to control grafts indicates that nerve growth factor acts to induce overall/premature differentiation and maturation. The mechanism for this whether or not it is receptor-mediated and which cells are primarily affected by nerve growth factor is not yet known. PMID- 2549446 TI - Reappearance of miniature endplate potentials in frog neuromuscular junctions "silenced" by lanthanum ions. AB - Miniature endplate potentials and currents were recorded from frog sartorius muscle fibres. As expected from previous work, La3+ caused a very large increase in miniature endplate potential frequency, followed by a progressive decrease, ending in the virtually complete disappearance of miniature endplate potentials. Soon after its application, La3+ caused an increase in the amplitude of miniature endplate potentials due to a lengthening of the underlying miniature endplate currents. Analysis of membrane current fluctuations produced by acetylcholine showed that La3+ caused an increase in the lifetime of the channels opened by acetylcholine. After miniature endplate potentials had vanished following La3+ treatment, the muscles were placed in organ culture conditions to see if miniature endplate potentials would reappear. After several hours miniature endplate potential activity was resumed in about 25% of the endplates, and disappeared again a few hours later, presumably because of accelerated nerve terminal degeneration. Preliminary electron microscopic observations suggest that the recovery of miniature endplate potential activity was accompanied by re formation of synaptic vesicles in the motor endings. PMID- 2549447 TI - Differential effects of (D)- and (L)-homocysteic acid on the membrane potential of cat caudate neurons in situ. AB - The enantiomers of homocysteic acid have been applied by microiontophoresis to neurons of the cat caudate nucleus in situ. The (L)-enantiomer elicited a bursty firing pattern similar to the one caused by N-methyl-D-aspartate, but differing from the N-methyl-D-aspartate pattern inasmuch as (L)-homocysteate induced depolarization shifts were shorter and had a smaller amplitude. (L)-Homocysteate induced excitations could be strongly inhibited by the selective N-methyl-D aspartate antagonist 2-amino-7-phosphonoheptanoic acid but they were less sensitive to this antagonist than N-methyl-D-aspartate itself. (D)-Homocysteate elicited a more regular firing pattern similar to the one caused by non-N-methyl D-aspartate excitatory amino acids such as quisqualate. These excitations were only rarely inhibited by 2-amino-7-phosphonoheptanoic acid. Our results suggest that (L)-homocysteate, a transmitter candidate at central mammalian synapses, is a mixed excitatory amino acid agonist with a strong preference for N-methyl-D aspartate receptors in the cat caudate nucleus, while (D)-homocysteate has a predominant action at non-N-methyl-D-aspartate excitatory amino acid receptors. PMID- 2549448 TI - 5'-Nucleotidase and the mabQ113 antigen share a common distribution in the cerebellar cortex of the mouse. AB - The parasagittal distribution patterns in the mouse cerebellar cortex for 5' nucleotidase and the antigen for the Purkinje cell specific antibody mabQ113 are identical and thus suggestive of a common organizational schema for cerebellar cortical compartments. PMID- 2549449 TI - Heterogeneity of presynaptic muscarinic receptors involved in modulation of transmitter release. AB - In order to extend the characterization of muscarinic receptors at presynaptic sites their inhibitory effect on the stimulation-evoked release of [3H]noradrenaline and [3H]acetylcholine from different axon terminals was studied and the dissociation constants and potencies of different antagonists were estimated, in guinea-pig and rat. While oxotremorine reduced the release of [3H]acetylcholine and [3H]-noradrenaline in a concentration-dependent manner from different release sites (Auerbach plexus, noradrenergic neurons in the right atrium, cerebral cortex), McN-A 343, an M1 receptor agonist, enhanced their release evoked by field stimulation. When the inhibitory effect of oxotremorine on transmitter release was studied, pancuronium, pirenzepine and atropine were competitive antagonists of presynaptic muscarinic receptors located on the noradrenergic axon terminals of the atrium. While atropine and pirenzepine inhibited the muscarinic receptors of cholinergic axon terminals in the Auerbach plexus, pancuronium and gallamine had a very low affinity. Significant differences were found in the affinity constants of antagonists for muscarinic receptors located in the cholinergic axon terminals of Auerbach plexus and cerebral cortex, and noradrenergic axon terminals of the atrium. While atropine and pirenzepine exerted similar effects on these presynaptic sites, pancuronium, gallamine and (11-(2-[diethylamino)-methyl)-1-piperidinyl)acetyl)-5, 11-dihydro 6(1-pyrido(2,3-b)(1,4)-benzodiazepin-6-on) were much more effective on muscarinic receptors controlling acetylcholine release from the cerebral cortex and noradrenaline release from the heart. There was more than 100-fold (2.0 pA2 units) difference in affinities of these antagonists.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549450 TI - Double-blind study of true vs. sham plasma exchange in patients treated with immunosuppression for acute attacks of multiple sclerosis. AB - We enrolled 116 patients in a multicenter, randomized, double-blind controlled trial of an 8-week course of 11 plasma exchange (PE) treatments in exacerbations of MS. The control group received sham PE, and both groups received identical treatment with IM ACTH and oral cyclophosphamide. Serum IgG decreased in the PE and sham treatment groups by 76% versus 22% by treatment 5, and by 64% versus 14% by treatment 11. PE also produced significant reductions in IgA, IgM, C3, and fibrinogen. PE patients had moderately enhanced improvement at 2 weeks relative to the sham group. PE patients with relapsing/remitting disease had significantly enhanced improvement at 4 weeks and there was also an increased improvement at 12 months, although this latter effect disappeared when we analyzed relapsing/remitting patients as a separate subgroup. Life table analysis showed the median time to recover preattack disability status was shorter in PE- than in sham-treated relapsing/remitting patients (4 vs. 13 weeks), a result confirmed by raw disability status scores in which there was recovery to their average preattack disability score by 3 months. PE given with ACTH plus cyclophosphamide enhances recovery from an exacerbation of disease in relapsing/remitting patients, although we observed no clear long-term benefits. PMID- 2549451 TI - The role of thermography in the evaluation of lumbosacral radiculopathy. AB - We studied 27 normal subjects and 30 patients with low back pain to evaluate the diagnostic accuracy of thermography in the diagnosis of lumbosacral radiculopathy. Thermographic abnormality was defined as the presence of either interside temperature difference exceeding 3 standard deviations from the normal mean, or an abnormal heat pattern overlying the lumbosacral spine. In patients with clinically unequivocal radiculopathy, thermography and electrophysiologic study were similar in diagnostic sensitivity, and the 2 methods agreed on the presence or absence of abnormality in 71% of cases. However, the thermographic findings had limited localizing value. Relative limb warming was often seen in patients with acute denervation on EMG, and limb cooling in those with more chronic lesions, but the side of the root lesion could not be identified confidently by thermography alone. Moreover, thermographic abnormalities appeared not to follow a dermatomal distribution and failed to identify the clinical or electrophysiologic level of radiculopathy in most cases. Thus, the thermographic findings are nonspecific, of little diagnostic value, and of uncertain prognostic relevance. PMID- 2549452 TI - Evidence in a lethal infantile mitochondrial disease for a nuclear mutation affecting respiratory complexes I and IV. AB - A child died at 4 months of age of a lethal infantile mitochondrial disease associated with cardiomyopathy. Detailed pathologic evaluation of this patient revealed abnormalities in the striated muscle, smooth muscle, heart, and liver, but not the central nervous system. Biochemical analysis revealed a combined complex I and IV deficiency in skeletal muscle, heart, and liver, but not in kidney and brain. Analysis of mitochondrial translation products and mitochondrial DNA failed to detect any abnormality. Parallel studies on both parents were uniformly normal. These data support the hypothesis that this disease was the result of a nuclear DNA mutation in a developmental stage specific and tissue-specific oxidative phosphorylation-gene. PMID- 2549453 TI - Gustatory pain: a complication of carotid endarterectomy. AB - A unique pain syndrome, triggered by gustatory stimuli in the absence of anatomic obstruction of the parotid duct, complicated carotid endarterectomy. We documented asymmetric parotid vasodilatation and sympathetic dysfunction in cutaneous areas overlaying the gland. We postulate that damage to the sympathetic innervation of the parotid gland resulted in unopposed parasympathetically mediated glandular vasodilatation. Vasoactive intestinal polypeptide may have played a role in producing symptoms. A similar syndrome occurs in patients with acute pandysautonomia. PMID- 2549454 TI - [Importance of preoperative monitoring in a case of hypochloric and hypokalemic dehydration subsequent to upper intestinal occlusion, caused by biliary ileus]. AB - A personally observed case of biliary ileus triggers this description of its clinical features, blood chemical and diagnostic aspects and surgical treatment. Emphasis is placed on the importance of the preoperative rehydration and restoration of the electrolyte balance in patients with upper intestinal occlusion which reduces the risk involved in subsequent surgery. PMID- 2549455 TI - [Intralesional therapy using recombinant interferon alpha 2 B in lesions of the uterine cervix caused by human papilloma virus]. AB - A clinical study on 20 patients with uterine cervix Human Papilloma Virus (HPV) lesions treated with intralesional recombinant alpha 2b interferon (3 millions I.U.) three times a week for three weeks where possible, is presented. After one year from treatment, lesions were cured or stabilized in 18 Patients (90%); progression was observed in 2 patients (10%) for which reason traditional surgical therapy was performed. PMID- 2549456 TI - Renal effects of enalapril in hypertensive patients with glomerulonephritis. AB - Renal effects of enalapril maleate in ten hypertensive patients with glomerulonephritis were evaluated after 1 and 16 weeks of therapy. Systemic blood pressure decreased, glomerular filtration rate was not significantly changed, and sodium fractional excretion and renal plasma flow increased, whereas renal vascular resistances and filtration fraction decreased acutely at the end of the study. Proteinuria diminished, but no variations in qualitative pattern were observed. ACE inhibitors, promoting efferent rather than afferent arteriolar vasodilatation and reduction of glomerular permeability coefficient, may reduce glomerular capillary hypertension and the development of proteinuria. PMID- 2549457 TI - New glycosylpeptides with high antinociceptive activity. AB - The antinociceptive activity of two new synthetic glucoside and galactoside enkephalinamide analogues was studied. The effects produced by the new analogues were compared with those obtained with [D-Met2,Hyp5]enkephalinamide and with morphine. The analogues were injected into the fourth ventricle and intrathecally. Tail immersion and paw pressure behavioural tests were used to assess antinociception. One of the analogues studied, O1,5-[beta-D galactopyranosyl] [D-Met2,Hyp5]enkephalinamide appears to be 57,000 times more potent than morphine. PMID- 2549458 TI - Synaptic localization of striatal NMDA, quisqualate and kainate receptors. AB - Striatal binding of labeled glutamate to N-methyl-D-aspartate (NMDA) receptors, D,L-alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid (AMPA) to quisqualate receptors and kainate to kainate receptors was examined in rats which had received unilateral decortications or unilateral striatal quinolinic acid lesions. One week after decortication, there were no significant changes in NMDA, quisqualate or kainate receptors in the striatum ipsilateral to the lesion, when compared to the striatum contralateral to the lesion. In contrast, binding to NMDA receptors was reduced by 92%, to quisqualate receptors by 80% and to kainate receptors by 81% in striatum 3 months after quinolinic acid lesions. The reduction in NMDA receptor binding was significantly greater than the loss of quisqualate or kainate receptors. These results suggest that NMDA, quisqualate and kainate receptor recognition sites are located postsynaptically in the striatum. These results also have implications for the quinolinic acid model of Huntington's disease. PMID- 2549459 TI - Synaptic transmission between cultured rat hippocampal neurons is enhanced by activation of protein kinase-C. AB - Synaptic connections between rat hippocampal neurons were studied in dissociated cell culture. Activation of a cultured neuron by pulse application of glutamate could produce postsynaptic currents (PSCs) in other neurons in the culture dish. Activation of protein kinase C (PKC) by a phorbol ester caused an enhancement of the magnitude of the PSCs without affecting much the delay and decay time constant of the recorded PSCs. The increased reactivity to synaptic activation was not accompanied by a postsynaptic change in sensitivity to topical application of an excitatory amino acid, glutamate. A PKC inhibitor polymyxin B reduced the effects of the phorbol ester. It is suggested that PKC activation plays an important role in the regulation of release of neurotransmitters from cultured central neurons. PMID- 2549460 TI - Effect of magnesium on depression of the monosynaptic reflex induced by 2 chloroadenosine or hypoxia in the isolated spinal cord of neonatal rats. AB - Superfusion of the isolated spinal cord of neonatal rats (4-9 days postpartum) with physiological medium containing 2-chloroadenosine (2-CA) or anoxic medium (equilibrated with 95% N2-5% CO2) depressed the evoked monosynaptic reflex (MSR) recorded extracellularly from a ventral spinal root. The effectiveness of 2-CA or anoxic medium in depressing the MSR was significantly reduced when the concentration of Mg2+ in the physiological medium was lowered from 1.25 X 10(-3) M to zero. The absence of Mg2+ resulted in a 7-fold shift to the right of the concentration-response curve to 2-CA and a reduction in the maximal depression of the MSR from 100% to 65 +/- 4% (mean +/- S.E.M.) of control. A 10 min exposure to anoxic medium containing 1.25 X 10(-3) M Mg2+ decreased the amplitude of the MSR to 23 +/- 6% of control, whilst in zero Mg2+ a decrease to only 50 +/- 5% of control was observed. These data provide further evidence that the response to adenosine, at the A1-receptor, is sensitive to Mg2+ ion concentration and suggest that there is an absolute requirement for Mg2+ in order to obtain full expression of the adenosine effect. Furthermore, the data are consistent with the hypothesis that adenosine is an important mediator of hypoxia-induced depression of the evoked MSR in the spinal cord, and suggest a potential role for Mg2+ during or after exposure to hypoxia in altering the actions of adenosine on neuronal activity or synaptic events. PMID- 2549461 TI - Modulation of spontaneous motility by GABAA receptor antagonism in the guinea pig isolated ileum. AB - Rhythmic neurally mediated spontaneous contractions of the longitudinal muscle in the isolated ileum of the guinea pig, sensitive to tetrodotoxin and atropine, were depressed and most often abolished by the GABAA receptor antagonists, bicuculline methiodide, RU 5135, and picrotoxin, a Cl- -ionophore blocker, as well as by GABA desensitization. 3-Mercaptopropionic acid, known to prevent GABA release, also reduced these naturally occurring spontaneous contractions. All these strongly indicate a physiological involvement of endogenous GABA in the control of spontaneous rhythmic activity in the intestine. PMID- 2549462 TI - 5-HT3 receptors control dopamine release in the nucleus accumbens of freely moving rats. AB - ICS 205-930, a selective and potent 5-HT3 receptor antagonist applied either systemically, or locally into the ventral tegmental area, antagonized the stimulation of dopamine release in the nucleus accumbens, induced by the subcutaneous administration of morphine. These findings, obtained by the use of brain microdialysis in awake freely-moving rats, demonstrate in vivo a functional role of 5-HT3 receptors in the brain. Since stimulation of dopamine release in the nucleus accumbens is a prerequisite for the expression of the rewarding properties of morphine, its suppression by ICS 205-930 suggests a possible application of 5-HT3 receptor antagonists in the treatment of addiction. PMID- 2549463 TI - N-methyl-D-aspartate promotes the survival of cerebellar granule cells: pharmacological characterization. AB - The survival of cerebellar granule cells in culture is promoted by chronic exposure to N-methyl-D-aspartate (NMDA). The effect is due to the stimulation of 'conventional' NMDA receptor-ionophore complex: it is concentration dependent, voltage dependent and blocked by the selective antagonists D-2-amino-5 phosphonovalerate, D-2-amino-7-phosphonoheptanoate, dextromethorphan and (+)-5 methyl-10,11-dihydro-5H-dibenzo(a,d)cyclohepten-5,10-imin emaleate (MK 801). The most potent antagonist tested was MK-801. In contrast, non-selective antagonists, including kynurenate, were much less effective. Further, the trophic effect of NMDA is not reproduced by ibotenate or quinolinate at the concentration range tested. It could also be shown that glutamate released into the culture medium is responsible for limited cell survival in the absence of NMDA. PMID- 2549464 TI - Microglial cells around amyloid plaques in Alzheimer's disease express leucocyte adhesion molecules of the LFA-1 family. AB - Immunostaining for glycoproteins of the LFA-1 family (leucocyte function associated antigens) was demonstrated on cells in the corona around senile plaques in Alzheimer's disease (AD) and in small glial cells in the subcortex of patients with AD and controls. These cells, which are usually referred to as microglial cells, showed positive immunohistochemical staining with monoclonal antibodies directed against the alpha-chains of all 3 LFA-1 family members, i.e. LFA-1, iC3b-receptor and P150,95, as well as with a monoclonal antibody against the common beta-chain. In the corona a diffuse staining for a ligand of LFA-1, intercellular adhesion molecule (ICAM)-1, was found as well. It is suggested that these molecules of the LFA-1 family may have a function in the dynamics of neuritic degeneration and sprouting. PMID- 2549465 TI - MK801-induced antagonism of NMDA-preferring excitatory amino acid receptors in horizontal cells of the turtle retina. AB - Intracellular recordings were made from axon terminals of L-type horizontal cells in the turtle (Pseudemys scripta elegans) retina. Superfusion with Ringer's solution containing 3.0 mM N-methyl-D-aspartate (NMDA) or 0.2 mM kainic acid (KA) induced depolarization and reduction in the hyperpolarizing light responses of horizontal cells, consistent with an agonist effect of these excitatory amino acid (EAA) analogs on postsynaptic receptors. Delivery of 0.1 mM MK801, a selective blocker of NMDA-type EAA receptors, had no apparent effect on membrane potential or photoresponses, nor did it change the KA depolarization. Exposure of the retina to 3.0 mM NMDA following 0.1 mM MK801 always caused hyperpolarization of the horizontal cell and loss of light responses. Because MK801 is specific for NMDA-preferring receptors, we suggest that the reversal of the NMDA response to one of antagonism following MK801 is strong evidence for the presence of NMDA preferring EAA receptors in turtle horizontal cells. PMID- 2549466 TI - Cholinergic neuromodulation by endothelin in guinea pig ileum. AB - The effect of endothelin on cholinergic neuroeffector transmission in guinea pig ileum was investigated. Endothelin was shown to inhibit the nerve-induced contractions and concomitantly to increase the basal muscle tone. Furthermore, endothelin inhibited the nerve-induced release of [3H]acetylcholine whereas the contractile response to exogenous acetylcholine was enhanced. In conclusion, our findings suggest that endothelin is a modulator of cholinergic neuroeffector transmission in guinea pig ileum with possible action via both inhibitory prejunctional and stimulatory postjunctional mechanisms. PMID- 2549468 TI - Nerve endings from rat brain tissue release copper upon depolarization. A possible role in regulating neuronal excitability. AB - Membrane vesicles from rat cerebral cortex were prepared and the functional response of the GABAA receptor was followed by monitoring GABA-activated influx of the radiotracer 36Cl- ion. CuCl2 decreased GABA-activated 36Cl- influx into synaptosomal membrane vesicles. The effect of Cu2+ was concentration dependent (5 500 microM CuCl2) and occurred with saturating (1 mM) as well as low (30 microM) GABA concentrations. A similar inhibition of the responses to muscimol (30 microM) was also observed with 50 microM CuCl2. In addition, release of copper from cortical synaptosomes and median eminence was followed by atomic absorption technique. An increased release of copper into the extracellular space was observed upon depolarization with 50 mM K+. A minimal concentration of copper was estimated to be 100 microM in the synaptic cleft. These findings suggest that copper may play a role in regulating neuronal excitability. PMID- 2549467 TI - Cytochrome oxidase activity in the rat retina following unilateral thalamic lesion. AB - The present study has investigated the cytochrome oxidase (CO) activity in retinas of normal rats and rats which received central lesions at birth or young adult stage. The results show that a thalamic lesion which injured the retinal ganglion cell axons in young adult rats led to severe loss of CO activity in the ganglion, inner and outer plexiform layers in the retina contralateral to the lesion as compared to those of normal rats. In contrast, distinct CO-reactive bands and cells were clearly observed in corresponding laminae in retinas in which almost the entire population of retinal ganglion cells was eliminated by a neonatal thalamic lesion. These results indicate that retinal ganglion cells contribute significantly to the CO activity observed in the inner retinal laminae under normal but not under abnormal conditions, and suggest that considerable changes in the activity of the remaining neurons and possibly reorganization of neural circuitry within the retina in rats which received neonatal lesions has taken place, as revealed by CO histochemistry. PMID- 2549469 TI - Proopiomelanocortin-derived peptides in mice with motoneurone disease. AB - beta-Endorphin and alpha-melanotropin immunoreactivity was detected in motor nerves in histological sections of murine skeletal muscle, using the peroxidase antiperoxidase technique. In mice of the wobbler strain, which can inherit motoneurone disease, a significantly higher proportion of the intramuscular nerves in soleus, extensor digitorum longus and diaphragm muscles of the diseased mice were immunoreactive than in the corresponding muscles of their healthy littermates. PMID- 2549471 TI - Raphe-produced excitation of spinal cord motoneurons in the cat. AB - In all motoneurons examined, both flexor and extensor, raphe (pallidus) stimulation consistently produced an excitatory postsynaptic potential (EPSP). Upon interaction with a group Ia EPSP there was summation resulting in neuronal discharge. The raphe-induced EPSP also facilitated the initial segment somadendritic coupling and hence the motoneuron excitability. These data support an excitatory role for raphe-spinal fibers on cat hindlimb motoneurons. PMID- 2549470 TI - Frequency dependent inhibition of the nicotinic transmission by serotonin in vesical pelvic ganglia of the rabbit. AB - Intracellular recordings were made from neurons in rabbit vesical pelvic ganglia (VPG), in vitro. Increasing the frequency of preganglionic-nerve stimulations from 0.1-1 Hz to 10-20 Hz facilitated fast excitatory postsynaptic potentials (EPSPs), resulting in a generation of action potentials. Acetylcholine-induced response was not altered during the facilitation of the fast EPSP. Serotonin blocked action potentials elicited by preganglionic-nerve stimulations at 0.1 Hz, while it caused no blockade at 10 Hz. Serotonin may potentiate the feature of high-pass filter in transmission of VPG. Immunohistochemical study demonstrated serotonin-like varicose terminals in rabbit VPG. PMID- 2549473 TI - Interactions of the pyrethroid fenvalerate with nerve membrane sodium channels: temperature dependence and mechanism of depolarization. AB - Depolarization of nerve membranes is an important component of the mode of action of pyrethroids, and its negative temperature dependence parallels that of insecticidal activity. We studied the mechanism and temperature dependence of depolarization of crayfish giant axons by pyrethroids, using intracellular microelectrode and voltage clamp techniques. Membrane depolarization caused by tetramethrin and fenvalerate was greater at 10 degrees C than at 21 degrees C, and was reversible upon changing the temperature. Short-duration depolarizing pulses in voltage-clamped fenvalerate-treated axons induced prolonged sodium currents that are typical of other pyrethroids, but the decay of the tail current following repolarization was extremely slow, lasting several minutes at the large negative holding potential of -120 mV. At the normal resting potential, the tail current did not decay completely, and even without stimulation, a steady-state sodium current developed, which could account for the depolarization. The steady state current induced by fenvalerate at the resting potential was much larger at 8 degrees C than at 21 degrees C, accounting for the negative temperature dependence of the depolarization. The negative temperature dependence of the steady-state current seems to be due ultimately to the great stabilizing effect of low temperature on the open-modified channel. When the steady-state current was induced at the resting potential, hyperpolarization to more negative potentials caused it to decay with exactly the same time course as tail currents induced by short-duration depolarizing pulses, indicating that both types of currents are carried by identically-modified channels. The modified channels were shown to be inactivated very slowly at potentials more positive than - 100 mV, accounting for the limited depolarization observed in micro-electrode experiments. Even when applied directly to the internal face of the membrane, the effect of fenvalerate on the sodium channel developed slowly, taking more than 90 min to reach its final level. Fenvalerate did not significantly affect potassium currents. PMID- 2549472 TI - Effects of repeated treatment with haloperidol on rat striatal neutral endopeptidase, and on mu- and delta-opioid binding sites: comparison with chronic morphine and chronic kelatorphan. AB - Neutral endopeptidase (NEP) and mu- and delta-opioid receptor densities were measured in sections of rat striatum by in vitro radioautography with the selective ligands: 3 nM [3H]N-[(2RS)-3-hydroxyaminocarbonyl-2-benzyl-1 oxopropyl]glycine ([ 3H]HACBO-Gly), 3 nM [3H]Tyr-D.Ala-Gly-(NMe)Phe-Gly-ol ([ 3H]DAGO) and 3 nM [3H]Tyr-D-Thr-Gly-Phe-Leu-Thr ([ 3H]DTLET), respectively. Haloperidol treatment (2 mg/kg/day, i.p., 3 weeks), which has been reported to increase enkephalin levels in the striatum, induced a 23% decrease in striatal (posterior level A= +8.4-8.6 mm) NEP labeling (but no change of mu- and delta sites). In contrast, no change in NEP occurred after chronic morphine (40-160 mg/kg/day, s.c., 4 days) or kelatorphan (10 nmol/h, i.c.v., 7 days), a mixed inhibitor of enkephalin-degrading peptidases. PMID- 2549474 TI - Differences in response of chickens from two genetic lines to diisopropyl phosphorofluoridate. AB - Neurotoxicity of diisopropyl phosphorofluoridate (DFP) was examined at 85 weeks of age in hens of two lines selected for high (HA) and low (LA) antibody response to sheep erythrocytes. DFP was administered by subcutaneous injection in doses of 0.25, 0.50 and 1.00 mg/kg and hens were observed for cholinergic signs at 30 min and for delayed neuropathy 8 to 14 days post-administration. Toxicity to DFP increased in severity with the dose and genetic differences were present because hens of line HA were more sensitive to DFP than were those of line LA. HA hens also had lower A-esterase activities and higher heterophil-to-lymphocyte ratios. No line x treatment interaction was evident, however, for activities of neurotoxic esterase or brain cholinesterase measured 24 hr after DFP administration. PMID- 2549475 TI - Temporal relationship of blood-nerve barrier breakdown to the metabolic and morphologic alterations of tellurium neuropathy. AB - The appearance of endoneurial edema early in the evolution of tellurium neuropathy raises the possibility that a breakdown of the blood-nerve barrier (BNB) plays a role in the pathogenesis of the tellurium-induced demyelination. To investigate this possibility, we correlated the temporal onset of breakdown of the BNB with inhibition of cholesterol synthesis and ultrastructural abnormalities in sciatic nerve of weanling Long-Evans rats fed a diet containing 1.1% elemental tellurium. Permeability of the BNB was assessed with [125I] albumin and horseradish peroxidase (HRP); cholesterol synthesis was assessed by incubating segments of sciatic nerve in vitro with [1-14C]acetate. Cholesterol synthesis was severely inhibited and labeled squalene was accumulating in sciatic nerve at 12 hr of tellurium exposure. The permeability of the BNB progressively increased between 24 hr and 72 hr of tellurium exposure. Membrane-delimited vacuoles, lipid droplets and cytoplasmic excrescences appeared in myelinating Schwann cells at 24 hr; demyelinating axons appeared at 48 hr of tellurium exposure. These observations suggest that factors other than BNB breakdown and vasogenic endoneurial edema are responsible for the initial Schwann-cell injury in tellurium neuropathy. However, the early onset of BNB breakdown may have a synergistic role in the pathogenesis of tellurium-induced demyelination. PMID- 2549477 TI - Vitamin D receptor mutations and familial rickets. PMID- 2549476 TI - A model for open-close control of cation channels in the plasma membrane of retinal rod outer segments. AB - A model for open-close control of cation channels in the plasma membrane of retinal rod outer segments is presented. A channel is assumed to open when 3 cGMP molecules bind to it and close as soon as one of the 3 cGMP molecules is released from it. The calcium ion (divalent cation) is a modulator of the channel conductance. The channel conductance is low when Ca2+ binds to it, while it is high when it is free from Ca2+. From the above assumptions, the reaction scheme of channels with cGMP and Ca2+ is created and the fraction of channels in the open and closed states was calculated using equations for this scheme. The kinetic constants used in the model are estimated from the experimental results of many studies and from the theories. From this estimation, it was found that at the physiological concentrations of intracellular and extracellular Ca2+, almost all channels are bound with Ca2+ and are in the low conductance state. The present model accounts for the reported dose(cGMP)-response(membrane current or conductance) relationship, where the Hill coefficient decreases as the cGMP concentration increases. The dark-level cGMP concentration of 8.13 microM is estimated from the model. This is in good agreement with the reported values. Moreover, the model predicts the invariance of current noise at relatively low Ca2+ concentrations when the cGMP concentration is raised from the dark level to a saturation level. The dynamic properties (opening and closing actions) of the channels in the present model are also in good agreement with the reported observations. The burst mode opening and closing of a channel is predicted by the present model, and it was found that the number of openings in a burst is controlled by the forward and backward rate constants between a channel protein and cGMP molecules. The simulated waveform of a single channel is similar to the reported observations. PMID- 2549478 TI - Identifying neurologic complications of A.I.D.S. PMID- 2549479 TI - Helping survivors survive. PMID- 2549480 TI - 99Tcm-diastereomeric dimercaptosuccinic acid complexes: comparison of their biodistribution. AB - The biodistribution of two stereoisomeric 99Tcm complexes derived from diastereomeric dimercaptosuccinic acids (DMSA), 99Tcm-dl-DMSA and 99Tcm-meso DMSA, was examined in sarcoma 180 tumour-bearing mice. The biodistribution pattern of meso-complex was not significantly different from 99Tc(V)m-DMSA, a new tumour imaging agent, while the dl-complex did not show the osteotropic property observed in meso- and 99Tc(V)m-DMSA complexes. The ratio of tumour to blood was superior for the 99Tcm-dl-DMSA (2.68) over the 99Tcm-meso-DMSA (1.25) at 3 h, and the ratio was reversed at 6 h, 3.13 to 4.28. The results indicate that dl-complex is of importance for the possible use in tumour imaging. PMID- 2549481 TI - [The functioning of mammalian mitochondria injected into fish embryos]. AB - The possibility of mammalian mitochondria functioning in fish embryos has been studied. Suspension of mitochondria isolated from the mouse fibroblast B-82/cap (chloramphenicol-resistant) and B-82 (chloramphenicol sensitive) cell cultures, were injected into the fertilized loach eggs. These embryos with an artificially increased number of mouse mitochondria developed and lived till the larval stages. Activity of cytochrome oxidase in these embryos was 1.5-2 times that in the control several hours after the injection, decreased during development and reached the control level by the gastrula stage. If these embryos with artificially increased number of mouse mitochondria were incubated in presence of chloramphenicol, only embryos that contained mitochondria from chloramphenicol resistant cells survived, thus suggesting that the injected mitochondria do not degrade but are preserved and function in the cytoplasm of developing loach embryos. PMID- 2549482 TI - [Effect of neonatal 6-hydroxydopamine administration on the catecholaminergic structures of the hypothalamus and on the behavior of rats of various ages]. AB - A neurotoxin 6-hydroxydopamine (6-OHDA) was introduced to 1, 2 and 3 day old male Wistar rats. Falck-Hillarp histochemical fluorescent method was used to analyze hypothalamic and hypophysial structures containing catecholamines. Statistically reliable decrease in catecholamine fluorescence intensity in hypothalamus and hypophysis, accumulation of catecholamines in nerve cells of supraoptic nucleus and in tanycytes of median eminence differed in 25- and 90-days rats. Disturbance of catecholaminergic systems of hypothalamus and hypophysis induced by neonatal introduction of 6-OHDA leads to reliable relaxation of orienting reaction and deep violation of learning that becomes stronger with age. PMID- 2549484 TI - Comparison of direct rapid tests for the detection of adenovirus antigen in routine conjunctival specimens. AB - Two direct rapid tests to detect adenovirus antigen, enzyme immunoassay (EIA) and immunofiltration (IF) were compared with regard to sensitivity, specificity, ease of interpretation, and technical complexity against 75 adenovirus culture positive and 35 adenovirus culture-negative conjunctival swab specimens. Enzyme immunoassay and IF were equally sensitive (P = 0.5), with sensitivities of 80.6 and 79.0%, respectively, when swab specimens were collected from patients within 7 days of the onset of clinical symptoms of adenovirus infection. After 7 days of clinical onset, IF (46.2%) was more sensitive (P less than 0.01) than EIA (0.0%), and was able to detect residual antigen in three culture-negative specimens. Enzyme immunoassay and IF were equally specific (100%). Both tests were easy to interpret, but IF was more technically complex and is not yet commercially packaged. Enzyme immunoassay could benefit either an office practice or a central laboratory, whereas IF is better suited for the latter. PMID- 2549483 TI - Ocular manifestations of acquired immune deficiency syndrome. AB - The ocular complications of acquired immune deficiency syndrome (AIDS) include: (1) a noninfectious microangiopathy, most often seen in the retina, consisting of cotton-wool spots with or without intraretinal hemorrhages and other microvascular abnormalities; (2) opportunistic ocular infections, primarily cytomegalovirus (CMV) retinitis; (3) conjunctival, eyelid, or orbital involvement by those neoplasms seen in patients with AIDS (i.e., Kaposi's sarcoma and lymphoma); and (4) neuro-ophthalmic lesions. In a series of 200 AIDS patients evaluated clinically, AIDS retinopathy was present in 66.5%. Sixty-four percent had cotton-wool spots, and 12% had intraretinal hemorrhages. Cytomegalovirus retinitis was diagnosed in 28% of AIDS patients. Neuro-ophthalmic lesions were found in 8% of all AIDS patients and were present in 33% of those patients with cryptococcal meningitis. Acquired immune deficiency syndrome retinopathy was present in 40% of 35 patients with the AIDS-related complex (ARC) and in 1.3% of 232 patients with asymptomatic human immunodeficiency virus (HIV) infection, evaluated photographically. These results suggest that the prevalence of AIDS retinopathy increases with increasing severity of HIV infection, and that CMV retinitis presents a significant vision-threatening problem in AIDS patients. PMID- 2549485 TI - Virus-associated dysplasia (bowenoid papulosis?) of the oral cavity. AB - We are presenting a case of multifocal, virus-associated dysplasia of the oral cavity. We believe this case represents bowenoid papulosis, which usually is limited to the genital region. The patient, a 21-year-old white man, had recently completed therapy for Hodgkin's disease. An oral examination revealed multiple red 3 to 6 mm macules scattered over the oral mucosa, involving the buccal and labial mucosa, palate, and gingiva. The favored clinical diagnosis was candidiasis. Histologically, the biopsy specimen showed severe epithelial dysplasia. Three additional oral biopsies of different sites were performed and revealed similar histology. Immunohistochemical stains for human papillomavirus were done, and two of four lesions stained positively. Transmission electron microscopy revealed intranuclear viral particles consistent with human papillomavirus. Further questioning and examination of the patient revealed that he had lesions of the penis that were clinically and histologically bowenoid papulosis. In addition, he admitted to oral-genital sex during the period of therapy for Hodgkin's disease. This is the first reported case of oral bowenoid papulosis, and it supports a viral cause for this disease process. PMID- 2549486 TI - [Malignant fibrous histiocytoma of the breast]. AB - In connection with the mammary tumor of a 41-year-old woman the authors draw attention to the rare occurrence of malignant fibrous histiocytoma in the mamma. The histology of the mammary malignant fibrous histiocytoma, role of immunohistochemical reactions in the diagnosis are described. The literature dealing with the malignant fibrous histiocytoma in the mamma is reviewed. This is the 11th case reported in the literature. The authors discuss the possibilities of the therapy of the malignant fibrous histiocytoma in the mamma. PMID- 2549487 TI - Acceleration of B-lymphoid tumorigenesis in E mu-myc transgenic mice by v-H-ras and v-raf but not v-abl. AB - Emu-myc transgenic mice constitutively express the c-myc oncogene from before birth, but their inexorable development of B-lymphoid tumors appears to require rare spontaneous alterations within B-lineage cells. To determine whether other oncogenes can cooperate with myc to induce B-lymphoid transformation, we infected neonatal Emu-myc mice and normal littermates with helper-free retroviruses bearing the v-H-ras, v-raf, or v-abl oncogene. The v-H-ras and the v-raf oncogene dramatically accelerated Emu-myc pre-B-lymphoma onset, whereas v-abl did not, despite the increased numbers of presumptive target pre-B cells in Emu-myc mice. These results imply that v-H-ras and v-raf synergize with deregulated myc expression to transform pre-B cells, while v-abl apparently does not. PMID- 2549488 TI - Characterization of alternate and truncated forms of murine c-myb proteins. AB - The expression of c-myb proteins in transformed and normal murine cells was investigated. Two c-myb proteins, p75c-myb and p90c-myb, were detected in normal thymocytes and cell lines with intact c-myb genes. These most likely differ by the inclusion of additional amino acids encoded by an alternatively spliced c-myb mRNA. The use of this alternative exon is therefore not a feature exclusively of those cells with viral integrations in their c-myb gene. Smaller c-myb proteins in myeloid leukemic cell lines with rearranged c-myb genes were also characterized. Viral integration into the 5' region of the c-myb gene in the W265 and W274 cell lines leads to the synthesis in each case of two amino-terminally truncated c-myb proteins. By contrast, in NFS60 cells, viral integration into a more 3' region of the c-myb locus (but upstream of an alternate exon) leads to the production of a single p50c-myb protein. PMID- 2549489 TI - Phosphatidylinositol kinase type I activity associates with various oncogene products. AB - We have assayed immunoprecipitates of several oncogene products from retrovirally infected chicken embryo fibroblasts (CEF) for phosphatidylinositol (PI) kinase activity. Immunoprecipitates of P68gag-ros, P130gag-tps, P47gag-crk, polyoma middle T (mT)-p60c-src complex, and mT-p62c-yrs complex exhibited PI kinase activity when assayed without detergents. This activity was sensitive to the nonionic detergent, Triton X-100, and the product was indistinguishable from phosphatidylinositol-3-phosphate, the product of kinase type I. Immunoprecipitates of p21Ha-ros protein did not contain any PI kinase type I activity. It has been suggested that an 81 kD protein phosphorylated in in vitro kinase assays of immunoprecipitates from mT-transformed rodent cells is responsible for the PI kinase type I activity seen in these immunoprecipitates. We have detected a chicken homologue of this 81 kD protein in immunoprecipitates of lysates from mT-transformed CEF. However, the chicken 81 kD protein sedimented more quickly than the PI kinase activity in sucrose gradients. In addition, the 81 kD protein was not detectable in protein kinase assays of immunoprecipitates of P68gag-ros or P130gag-fps. These results suggest that the 81 kD protein may not be the PI kinase. PMID- 2549491 TI - Adenoid cystic carcinoma of the breast. A morphologically heterogeneous neoplasm. PMID- 2549490 TI - Viral growth, origin binding, and p53 binding properties of simian virus 40 large T antigen transformation and replication mutants. AB - The viability, p53 binding, and SV40 origin binding of a series of SV40 large T antigen point mutants, which map to the amino terminal one-third of the molecule, were examined. Two mutants which yield small plaques were found to have altered kinetics of replication upon infection of permissive cells. Mutants which did not bind to the origin of replication were not able to replicate, but the reverse was not always true. Replication defective mutants which bound the SV40 origin were found; these map both inside and outside of the origin binding domain. All the transformation defective mutants bound the cellular protein, p53. PMID- 2549493 TI - Replication of herpes simplex virus in blood monocytes and placental macrophages from human neonates. AB - Increased permissiveness of macrophages for herpes simplex virus (HSV) replication may be a mechanism for the dissemination and severity of neonatal herpetic infection. We have assessed the replication of HSV in neonatal blood monocytes and placental macrophages using several criteria for viral permissiveness. Assay of production of infectious progeny virus indicated that cord blood monocytes, like adult monocytes, were nonpermissive for HSV (about 1% of cells producing virus). In vitro culture of cord blood monocytes resulted in increased replication of HSV, but no greater extent than virus production in cultured adult cells. HSV infection of fetal placental macrophages was weak but present (4.4% of cells). Assay of production of viral antigens and electron microscopic analysis of structural elements indicated that a larger number of cord blood monocytes and placental macrophages were abortively infected than were productively infected. These results indicate that monocytes and macrophages from human neonates do not show the enhanced permissiveness for HSV demonstrated in newborn mice and suggest that dissemination of herpetic infection in human newborns cannot be explained by increased neonatal monocyte permissiveness for HSV. PMID- 2549492 TI - The effects of recombinant human granulocyte-macrophage colony stimulating factor on in vitro cord blood granulocyte function. AB - Neonatal granulocytes are recognized to have functional defects which are thought to be important in the increased susceptibility to infection in the neonate. Recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF), a member of a family of glycoproteins essential for the in vitro survival, proliferation, and differentiation of hematopoietic progenitor cells, has been shown to enhance the functional capabilities of adult granulocytes. This study tested the effects of rhGM-CSF on the locomotion, superoxide generation, phagocytosis, bactericidal activity, and membrane depolarization responses of cord blood granulocytes. Concentrations of rhGM-CSF between 100 and 1 pM significantly enhanced the leading front of cord blood granulocyte locomotion. The mean distance migrated by the cell population and the number of cells responding to the chemoattractant were also significantly enhanced in cord blood granulocytes treated with 1 pM rhGM-CSF. Superoxide anion production was significantly enhanced in cord blood granulocytes stimulated with fMLP after a 30 or 60-min exposure to 100 pM rhGM-CSF. However, this enhancing effect was not observed in cells incubated with rhGM-CSF for 2 h before formyl-methionine leucine-phenylalanine stimulation. Phagocytosis, bactericidal activity, and membrane depolarization responses of cord blood granulocytes were not affected by exposure to rhGM-CSF. These findings demonstrate that selected cord blood granulocyte functions are enhanced by in vitro exposure to rhGM-CSF. Whether these in vitro observations have in vivo significance await further study. PMID- 2549494 TI - Outbreak of herpangina associated with Coxsackievirus B3 infection. AB - An outbreak of acute febrile illness was observed in summer, 1987, in a welfare home in which 31 healthy infants were accommodated. Within a 5-day period 25 infants (81%) acquired a febrile illness. Coxsackievirus B3 was isolated from 16 (64%) of 25 throat swabs. In the patients in whom viral culture was negative or not performed, 6 were serologically identified as having a coxsackievirus B3 infection. Among 22 patients identified as having a coxsackievirus B3 infection 7 had typical herpangina and the others had pharyngitis with or without a few small vesicles. Serum alpha-interferon was detected in all but 2 cases (one with proved infection and another with indefinite infection). Herpangina can be associated with coxsackievirus B3 as well as with the more frequently associated coxsackievirus Group A; this explosive type of outbreak might be transmitted by a small particle aerosol. PMID- 2549496 TI - Serologic misdiagnosis of congenital infections. PMID- 2549495 TI - Association of parainfluenza virus type 3 infection with allograft rejection in a liver transplant recipient. PMID- 2549497 TI - Viral enteritis in the 1980s: perspective, diagnosis and outlook for prevention. PMID- 2549498 TI - Humoral immunity and clinical reinfections following varicella vaccine in healthy children. AB - The duration of immunity following varicella-zoster vaccination in healthy children remains a critical issue. In a 3-year study of 140 OKA/Merck vaccine recipients, duration of immunity was assessed by two measures. The first was persistence of varicella-zoster antibody measured by modified fluorescent antibody to membrane antigen test. Thirty-six toddler vaccinees 12 to 24 months of age had sera obtained at 6 weeks, 1 year, and 2 years. Geometric mean titer +/ SD at 6 weeks was 57.7 +/- 2.9; at 1 year, it was 12.4 +/- 3.9; at 2 years, it was 9.9 +/- 3.9. Repeated-measures analysis of variance showed a significant overall decrease in antibody titer with time (F = 30.62, P less than .001). Post hoc comparisons indicated that the 6-week and 1-year titers were significantly different (P less than .001), but the difference between 1 and 2 years was not (P = .138). Clinical reinfections were also examined for 3 years after vaccination. Suspected varicella cases were confirmed by a fourfold or more increase in titer. Of 84 toddlers, 68 were exposed one or more times, and 6 became reinfected. Of 49 siblings, 45 were exposed, and one became reinfected. All 7 children had less than 70 vesicles and 6 of 7 were afebrile. No cases of zoster occurred. It is concluded that OKA/Merck varicella-zoster vaccination leads to antibody persistence for 2 years and the few reinfections that do occur are greatly attenuated. PMID- 2549499 TI - [Diagnostic possibilities of static gamma-scintigraphy using 99mTc pyrophosphate and Tc 99m phosphate in children with glomerulo nephritis]. AB - Children with nephrotic glomerulonephritis were examined for incorporation of 99mTc-pyrophosphate and 99mTc-phosphone. The relationship was discovered between the accumulation of the radiopharmaceutical drug and the sensitivity to glucocorticoids used in the pathogenetic treatment. Accumulation of the drug in the course of the treatment documents the presence of complete, partial remission or the disease activity. It is recommended that static gammascintigraphy should be carried out for early assessment of the efficacy of the treatment and its correction. PMID- 2549500 TI - Treatment of chronic pain syndromes with transcutaneous iontophoresis of vinca alkaloids, with special regard to post-herpetic neuralgia. AB - Successful treatment of 35 post-herpetic neuralgia (PHN) patients by means of transcutaneous iontophoresis of Vincristine is reported. This technique, based on transganglionic regulation--a novel neurobiological principle discovered by Csillik and Knyihar-Csillik-, alleviated pain in both fresh and inveterated PHN cases. Statistical analysis of the results excludes a placebo-like action. PMID- 2549501 TI - The use of clone-specific anti-VJ junction oligonucleotides to identify minor clonal lymphoid populations. AB - The polymerase chain reaction was used to amplify rearrangements of T-cell receptor gamma genes from 2 leukemic DNA samples involving the same variable segment. One V-J junction was sequenced and an anti-junctional oligonucleotide synthesized. This oligonucleotide was used as probe on in vitro amplified DNA. We show that this strategy allows the detection of the corresponding clonal DNA at dilutions of up to 10(6) fold in germline DNA. Moreover we demonstrate that it is possible to differentiate this clone from the second leukemic clone and from polyclonal T-cells. PMID- 2549502 TI - The impact of EBV hybridization in clinical and biological medicine. AB - Hybridization technology allows the detection of EBV (Epstein-Barr Virus) DNA in lymphoproliferative diseases in immunosuppressed patients. Moreover, in HIV infected patients, LIP (Lymphocytic Interstitial Pneumonitis), hairy leucoplasia and lymphoma are associated with EBV by DNA hybridization. These procedures are also useful for understanding interactions between EBV and epithelial cells. PMID- 2549503 TI - The effect of specific mutations at and around the gag-pol gene junction of Moloney murine leukaemia virus. AB - By carrying out oligonucleotide-directed mutagenesis, in vitro, on a 3.3 kb XhoI HindIII fragment from Moloney murine leukaemia virus Mo-MuLV proviral DNA, inserted into the phagemid pTZ19R, nine separate fragments have been prepared in which mutations have been inserted at and around the gag-pol gene junction. Using these mutant fragments Mo-MuLV proviral DNA has been reassembled and cloned into pBR322. Examination of the mutant proviral DNAs in mouse culture cells indicates that a terminator codon at the gag-pol junction is essential for function, but any of the three chain terminator codons gives an active virus. Also the region of secondary structure surrounding the terminator codon must be preserved. PMID- 2549504 TI - Site-specific location of covalent DNA-polypeptide complexes in the chicken genome. AB - A nitrocellulose filter binding assay was applied to isolate and to analyze the fraction of chicken DNA fragments associated with residual nuclear polypeptides resistant to SDS/proteinase K treatment and phenol extraction. It is shown that the DNA-polypeptide complexes retained on nitrocellulose filters are located on a non-random sub-set of DNA sequences. (a) Southern analysis reveals that the fractions of DNA fragments from chicken erythrocytes and from hen oviduct cells associated with the resistant polypeptides have a lower sequence complexity than unfractionated DNA. Moreover, the retained DNA fractions from different cell types of the same species are highly homologous. (b) All DNA fragments of the transcriptionally active and inactive ovalbumin gene map in the DNA fraction passing the filters indicating that the tight DNA-polypeptide complexes are not remnants of transcription complexes. (c) By use of a genomic sub-set library prepared from DNA retained on filters, clones were isolated with sequences mapping specifically in the DNA fraction associated with the tight DNA polypeptide complexes. The results are consistent with fixed covalent DNA polypeptide complexes in the chicken genome whose location is essentially identical in different cell types of the same species and apparently determined by DNA signal-sequences. PMID- 2549505 TI - Replication of wheat dwarf virus DNA in protoplasts and analysis of coat protein mutants in protoplasts and plants. AB - The replication of wheat dwarf virus (WDV) in protoplasts derived from a Triticum monococcum suspension cell system was investigated. The production of circular viral double-stranded DNA (dsDNA) forms consistent with the replication of the viral genome was observed. In comparison to whole plants, the production of viral single-stranded DNA (ssDNA) was reduced, possibly due to only low levels of viral coat protein being produced in the protoplasts. Mutations introduced into the viral coat protein open reading frame (ORF) did not affect the ability of the viral DNA to replicate, and a deletion of ca. 400 bp was tolerated. However, these mutations abolished the infectivity of the viral genome when agroinoculated onto wheat plants, providing evidence that, contrary to the case for the bipartite geminiviruses, the coat protein is essential for infection by WDV. PMID- 2549506 TI - The recombinational enhancer for DNA inversion functions independent of its orientation as a consequence of dyad symmetry in the Fis-DNA complex. AB - The Escherichia coli Fis protein binds to specific DNA sequences whose base composition varies enormously. One known function of Fis is to stimulate site specific DNA recombination. We used the Gin-mediated DNA inversion system of bacteriophage Mu to analyze Fis-DNA interaction. Efficient inversion requires an enhancer which consists of two Fis binding sites at a fixed distance from each other. Using mutant enhancers in which one of the Fis binding sites is replaced we show that Fis binds symmetrically to the DNA and we locate the center of symmetry. Furthermore, we show that one of the Fis binding sites can be replaced by a Fis binding site that normally functions in a process other than site specific recombination. PMID- 2549508 TI - Thermodynamic stability and statistical significance of potential stem-loop structures situated at the frameshift sites of retroviruses. AB - RNA stem-loop structures situated just 3' to the frameshift sites of the retroviral gag-pol or gag-pro and pro-pol regions may make important contributions to frame-shifting in retroviruses. In this study, the thermodynamic stability and statistical significance of such secondary structural features relative to others in the sequence have been assessed using a newly developed method that combines calculations of the lowest free energy of formation of RNA secondary structures and the Monte Carlo simulations. Our results show that stem loop structures situated just 3' to the frameshift sites are both highly stable and statistically significant relative to others in the gag-pol or gag-pro and pro-pol junction domains (both 300 nucleotides upstream and downstream from the possible frameshift sites are included) of Rous sarcoma virus (RSV), human immunodeficiency virus (HIV-1), bovine leukemia virus (BLV), human T-cell leukemia virus type II (HTLV-II), and mouse mammary tumor virus (MMTV). No other more stable, or significant folding regions are predicted in these domains. PMID- 2549507 TI - Isolation and characterization of a human telomere. AB - A method is described that allows cloning of human telomeres in S. cerevisiae by joining human telomeric restriction fragments to yeast artificial chromosome halves. The resulting chimeric yeast-human chromosomes propagate as true linear chromosomes, demonstrating that the human telomere structure is capable of functioning in yeast and suggesting that telomere functions are evolutionarily conserved between yeast and human. One cloned human telomere, yHT1, contains 4 kb of human genomic DNA sequence next to the tandemly repeating TTAGGG hexanucleotide. Genomic hybridizations using both cloned DNA and TTAGGG repeats have revealed a common structural organization of human telomeres. This 4 kb of genomic DNA sequence is present in most, but not all, human telomeres, suggesting that the region is not involved in crucial chromosome-specific functions. However, the extent of common features among the human telomeres and possible similarities in organization with yeast telomeres suggest that this region may play a role in general chromosome behavior such as telomere-telomere interactions. Unlike the simple telomeric TTAGGG repeats, our cloned human genomic DNA sequence does not cross-hybridize with rodent DNA. Thus, this clone allows the identifications of the terminal restriction fragments of specific human chromosomes in human-rodent hybrid cells. PMID- 2549509 TI - Ubiquitin expression in Neurospora crassa: cloning and sequencing of a polyubiquitin gene. AB - We have cloned and sequenced a polyubiquitin gene from Neurospora crassa that is organized in a four repeat-tandem array. The first repeat contains a small intron and the last is fused to an extra glutamine codon. In Northern blots, two RNA species of 1.3 kb and 0.7 kb hybridize to the isolated clone. The larger ubiquitin (UBI) transcript accumulates after partial inhibition of protein synthesis with cycloheximide, and the smaller one preferentially accumulates in conidia after germination. Unexpectedly, constitutive expression of UBI transcripts in exponentially grown mycelia is not altered by heat-shock or exposure to arsenite. PMID- 2549510 TI - Cloning and characterization of mutL and mutS genes of Vibrio cholerae: nucleotide sequence of the mutL gene. AB - The mutL and mutS genes of Vibrio cholerae have been identified using interspecific complementation of Escherichia coli mutL and mutS mutants with plasmids containing the gene bank of V. cholerae. The recombinant plasmid pJT470, containing a 4.7 kb fragment of V. cholerae DNA codes for a protein of molecular weight 92,000. The product of this gene reduces the spontaneous mutation frequency of the E. coli mutS mutant. The plasmid, designated pJT250, containing a 2.5 kb DNA fragment of V. cholerae and coding for a protein of molecular weight 62,000, complements the mutL gene function of E. coli mutL mutants. These gene products are involved in the repair of mismatches in DNA. The complete nucleotide sequence of mutL gene of V. cholerae has been determined. PMID- 2549511 TI - Structure and expression of a muscle specific gene which is adjacent to the Drosophila myosin heavy-chain gene and can encode a cytochrome b related protein. AB - We report the characterization of a transcription unit at the second chromosome locus 36B, designated TU-36B which is adjacent to the 3' end of the Drosophila myosin heavy-chain (Mhc) gene. We have isolated and sequenced a complementary DNA clone and the region of genomic DNA which represents this gene. The sequencing studies reveal that this gene contains one intron, the mRNA is 1480 nucleotides in length, the TU-36B mRNA is transcribed in an orientation opposite to the Mhc mRNAs, and the poly(A) addition site of this gene is 99 nucleotides downstream of poly(A) addition site A-2 of the Drosophila Mhc gene. The mRNA contains a continuous open reading frame which can encode a protein product of 47,000 daltons in molecular weight. The proposed protein shares homology with cytochrome b proteins. Comparison of in situ hybridization of Mhc specific and TU-36B specific probes to tissue sections demonstrates that both mRNAs are predominantly transcribed in the same muscle tissues of the developing fly. PMID- 2549512 TI - Nucleotide sequence of cDNA for rat liver and brain cytochrome c oxidase subunit VIa (Vb). PMID- 2549514 TI - Porcine OSU rotavirus segment II sequence shows common features with the viral gene of human origin. PMID- 2549513 TI - The nucleotide sequence of env gene of duck-cells adapted Rous sarcoma virus. PMID- 2549516 TI - Nucleotide sequence of a cDNA for bovine cytochrome c oxidase subunit VIIa. PMID- 2549515 TI - Sequence of a cDNA specifying subunit VIa of human cytochrome c oxidase. PMID- 2549517 TI - The nucleotide sequence of the 3' region of the murine leukemia virus SL16c4. PMID- 2549518 TI - Eukaryotic expression vectors for the analysis of mutant proteins. PMID- 2549520 TI - Body politic--the parting of the ways? PMID- 2549519 TI - Surgical care of the elderly. Home help. PMID- 2549521 TI - Orchestrating the process of patient education. Methods and approaches. AB - Patient education is a must, yet many obstacles prevent the achievement of this important goal. Nurses need to examine methods and approaches that have application in today's practice. These include the nurse's assessment of the patient as well as a self-inventory so a multifaceted plan can be implemented to provide information during each patient care contact point. Suggested methods, such as contracting and use of computers in the education process, are additional ways to involve patients actively so they can maximize their fullest potential in self-care. PMID- 2549522 TI - Realistically meeting the educational needs of hospitalized acute and short-stay patients. AB - Patient teaching in acute care hospital settings isn't much different from teaching in other settings. The main differences have to do with the amount of time available and the condition of the patients. Using good assessment practices, sticking to "need to know" survival content, simplifying instructions, streamlining programs, providing for practice, providing reference and resource materials, and referring patients for further education are the ingredients that will help get the job done efficiently and effectively. PMID- 2549523 TI - Continuity of care. Balancing care of elders between health care settings. AB - Continuing care planning and balancing care of elders are crucial components of health care today. The most sweeping change that has come with the advent of prospective pricing for the acute care setting is in the financing of health care for the aged and disabled. Prospective pricing, which uses the diagnosis-related group system, is the method chosen by federal agencies to restrain Medicare costs. For almost two decades, the hospital has been the reservoir for the impaired elderly who could not be returned home or to the community without support services. The system now encourages shortened hospital stays. Continuing care planning is the key to balancing the care of the elder through the process of discharge planning. We must assure patients and families that their needs will be met in the community when the patient is discharged. Balancing care of elders between health care settings through discharge planning is identified in three simple words: A Complex Process. As health care professionals, we must be ready to accept this challenge. PMID- 2549524 TI - Radical resection of the shoulder girdle for a malignant tumor: four case reports. AB - Four cases of malignant tumor of the shoulder were treated by radical resection of the shoulder girdle, saving the upper extremity. During follow up, all patients were able to use their upper extremities with satisfactory function and stability. This article presents the indications, operative technique, and follow up. PMID- 2549525 TI - Nerve entrapments: an update. PMID- 2549526 TI - The absence of sex and age effects on the corticosterone response of turkey poults to adrenocorticotropic hormone and temperature stressors. AB - A study was undertaken to assess the role of sex and age on the adrenal cortical response of turkey poults during the time period associated with idiopathic poult mortality. Three and 14-day-old tom and hen turkey poults were subjected to the following treatments: saline injection (control), cool water immersion (20 C) for 1 min, cold water immersion (5 C) for 1 min, and 1 IU/kg adrenocorticotrophic hormone (ACTH) injection. Blood was collected 1 and 4 h posttreatment for plasma corticosterone (CS) assay. Temperature treatments did not affect plasma CS in either age or sex. Plasma CS was increased by ACTH after 4 h postinjection in both sexes and at both ages. The hypothalamic-pituitary-adrenal cortical axis of the turkey poult was functional but appeared to be insensitive to temperature stress. PMID- 2549528 TI - Relaxant effects of the selective phosphodiesterase inhibitors milrinone and OPC 3911 on isolated human mesenteric vessels. AB - Several new positive inotropic drugs with vasodilating properties for which a major mechanism of action is believed to be inhibition of phosphodiesterase (PDE) have been introduced in the treatment of congestive heart failure. Hydrolysis of cyclic nucleotides is catalyzed by multiple forms of PDE, which may vary between organs and cell-types. These enzymes can be selectively inhibited by various agents, theoretically making it possible to produce tissue-selective responses. An enzyme, which belongs to a subclass of cGMP inhibited low Km cAMP PDE, was recently purified from rat adipose tissue. The enzyme was specifically and potently inhibited by the cilostamide derivative OPC 3911 (OPC) and milrinone (mil). We studied the relaxant effects of OPC and mil on isolated human mesenteric arteries and veins in vitro. In preparations contracted by noradrenaline (NA), both agents produced about 60% maximum relaxation; OPC was 3 4 times more potent than mil. Both OPC and mil caused rightward displacement of the NA concentration-response curve and depressed the maximum responses. Arteries, as compared to veins, were more sensitive to this inhibition of NA contraction. Both drugs relaxed arteries contracted by 30 mM K+, but not 127 mM K+; maximum relaxation was between 60 and 70%. OPC was 10 times more potent than mil. The interactions between mil/OPC and isoprenaline, forskolin and ouabain were also studied. In preparations pretreated with isoprenaline or forskolin, the relaxant effects of mil and OPC were additive to those of isoprenaline and forskolin. Ouabain pretreatment did not affect the actions of the phosphodiesterase inhibitors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549527 TI - 3H-muscimol binding sites within the rat choroid plexus: pharmacological characterization and autoradiographic localization. AB - By using combined radioreceptor binding and autoradiographic techniques, we were able to localize the GABA 'A' receptor agonist 3H-muscimol in the rat choroid plexus. Within sections of rat choroid plexus, 3H-muscimol was bound specifically with a KD of 37 nM and a Bmax of 253 pmol/mg tissue. These values are consistent with the labelling of GABA 'A' receptor sites. The autoradiographic studies demonstrated that 3H-muscimol was attached to the epithelium of the choroid plexus. The blood vessels of the plexus did not exhibit specific labelling. Examination of these data suggests the existence of GABAergic mechanisms which control cerebrospinal fluid production or flow. PMID- 2549529 TI - Chronotropic responses in rats implanted with an acrylic orthopaedic cement. AB - The chronotropic and inotropic responses were studied in isolated atria and papillary muscles from rats who had carried a pellet of methyl methacrylate polymer for one week and from sham operated control rats. Right atria isolated from implanted rats were hyporesponsive to the positive chronotropic effects of isoproterenol and glucagon but not to dibutyryl cAMP or to CaC12. Inotropic response of left atria and papillary muscles to isoproterenol was unaltered. No change in the beta-receptor density or affinity was associated with these phenomena. The results indicate that methyl methacrylate exposure leads to a disruption in the signal transduction of chronotropic stimuli mediated by the adenylate cyclase system. The alteration occurs at some point after receptor binding and before cAMP action. PMID- 2549530 TI - Long-term effect of antidepressant drugs and electroconvulsive shock (ECS) on cortical alpha 1-adrenoceptors following destruction of dopaminergic nerve terminals. AB - The effect of repeated treatment with imipramine, citalopram and ECS on the density of alpha 1-adrenoceptors in the cerebral cortex of rats with a dopaminergic lesion was studied. Imipramine and citalopram produced alph 1 upregulation in normal animals but not in animals with a dopaminergic lesion. On the other hand, ECS, produced such an effect in both normal and lesioned rats. Our results indicate that the investigated drugs (imipramine, citalopram) and ECS induce alpha 1-up-regulation via different mechanisms, and that the effect of these drugs depends on intact dopaminergic nerve terminals. PMID- 2549531 TI - Investigation of the regulation of pigmentation in alpha-melanocyte-stimulating hormone responsive and unresponsive cultured B16 melanoma cells. AB - In vitro melanocyte-stimulating hormone (MSH) stimulates melanogenesis in some, but not all, melanocytes and melanoma cells. In an attempt to explain this variation in response to alpha MSH, we examined cyclic adenosine monophosphate (cAMP) accumulation, tyrosinase activity, and melanin production in primary (1 degree) murine B16 melanoma cells and in two B16 cell lines (B16 F1 and B16 F10) that are known to respond to alpha MSH. In vivo all three B16 melanoma cell types produced pigmented tumours. In vitro alpha MSH increased tyrosinase activity and melanin content in the F1 and F10 cells but not in the B16 1 degree cells. alpha MSH, however, increased cAMP production in all three cell types, confirming that the inability of B16 1 degree cells to produce melanin in response to alpha MSH is not due to a lack of alpha MSH receptors or cAMP response to alpha MSH. Further, we present evidence for a separate pathway of melanogenesis that is independent of cAMP as calmodulin antagonists, which do not elevate cAMP, increased tyrosinase activity, and melanin production in both 1 degree and F1 cells. PMID- 2549532 TI - Control of melanoblast differentiation in amphibia by alpha-melanocyte stimulating hormone, a serum melanization factor, and a melanization inhibiting factor. AB - A ventrally localized melanization inhibiting factor (MIF) has been suggested to play an important role in the establishment of the dorsal-ventral pigment pattern in Xenopus laevis [Fukuzawa and Ide:Dev. Biol., 129:25-36, 1988]. To examine the possibility that melanoblast expression might be controlled by local putative MIF and melanogenic factors, the effects of alpha-melanocyte stimulating hormone (alpha-MSH), a serum melanization factor (SMF) from X. laevis or Rana pipiens, and MIF on the "outgrowth" and "melanization" of Xenopus neural crest cells were studied. Outgrowth represents the number of neural crest cells emigrating from cultured neural tubes, and melanization concerns the percentage of differentiated melanophores among the emigrated cells. MSH or SMF stimulate both outgrowth and melanization. The melanogenic effect of Xenopus serum in this system is more than twice that of Rana serum. The actions of MSH and Xenopus serum on melanization seem to be different: 1) Stronger melanization is induced by Xenopus serum than by MSH, and the onset of melanization occurs earlier with Xenopus serum; 2) MSH stimulates melanization only in the presence of added tyrosine; and 3) MSH causes young melanophores to assume a prominent state of melanophore dispersion during culture, while Xenopus serum (10%) had only a slight dispersing effect and not until day 3. A fraction of Xenopus serum presumably containing molecules of a smaller molecular weight (MW less than 30 kDa) than that of a pigment promoting factor reported in calf serum [Jerdan et al.: J. Cell Biol., 100:1493-1498, 1985] produces the same remarkable melanogenic effects as does intact serum. While this fraction stimulates outgrowth, another fraction presumably containing larger molecules (MW greater than 100 kDa) does not. MIF contained in Xenopus ventral skin conditioned medium (VCM) inhibits both outgrowth and melanization dose dependently. When VCM is used in combination with MSH, the stimulating effects of MSH on both outgrowth and melanization are completely inhibited. In contrast, the stimulatory effects of Xenopus serum are not completely inhibited when combined with VCM, although melanization is reduced to approximately 40% that of controls. MIF activity was also found to be present in ventral, but not in dorsal, skin conditioned media of R. pipiens when tested in the Xenopus neural crest system.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2549533 TI - Immunobiology of human infection by Entamoeba histolytica. PMID- 2549534 TI - [Synaptophysin, chromogranin A and neuron-specific enolase as tumor markers in neuroendocrine tumors of the gastrointestinal tract and lung. An immunohistochemical study]. PMID- 2549535 TI - [Protective effect of nedocromil on hypo- and hypertonic solution-induced airway obstruction in exogenous allergic bronchial asthma]. AB - Nedocromil is a new non-bronchodilator substance with mast cell stabilizing properties. In an acute placebo-controlled randomized double blind study, the protective effect of nedocromil on the bronchoconstrictor action of nebulised hypo-(distilled water) and hyperosmolar (4% saline) solutions was investigated in 12 patients with allergic asthma. Inhalations of distilled water and hypertonic saline had similar bronchoconstrictor effects. The effect of both solutions was significantly (p less than 0.01) attenuated by pretreatment with nedocromil, as compared with placebo. Thus, nedocromil could be useful in the treatment of allergic asthma. PMID- 2549536 TI - A stereological study of the differentiation process in Trypanosoma cruzi. AB - When epimastigote forms of Trypanosoma cruzi grown in a rich medium (LIT) are transferred to a simple, chemically defined medium (TAU3AAG, containing Ca2+, Mg2+, K+, Na+, L-proline, L-glutamate, and L-aspartate in phosphate buffer) they transform into trypomastigote forms. Morphometric analysis of transmission electron micrographs of thin sections of parasites collected at different steps of the transformation process showed that no changes occurred in the volume density of mitochondria and cytoplasmic vacuoles. However, a significant increase in the volume density of the kinetoplast DNA network as well as the lipid inclusions and a decrease in that of the reservosome (a special type of endosome) was observed. These observations suggest that during differentiation, T. cruzi accumulates lipids and uses molecules contained in the reservosome as its main energy source. PMID- 2549537 TI - Mechanism of oligonucleotide uptake by cells: involvement of specific receptors? AB - We have investigated the interaction of oligonucleotides and their alkylating derivatives with mammalian cells. In experiments with L929 mouse fibroblast and Krebs 2 ascites carcinoma cells, it was found that cellular uptake of oligodeoxynucleotide derivatives is achieved by an endocytosis mechanism. Uptake is considerably more efficient at low oligomer concentration (less than 1 microM), because at this concentration a significant percentage of the total oligomer pool is absorbed on the cell surface and internalized by a more efficient absorptive endocytosis process. Two modified proteins were detected in mouse fibroblasts that were treated with the alkylating oligonucleotide derivatives. The binding of the oligomers to the proteins is inhibited by other oligodeoxynucleotides, single- and double-stranded DNA, and RNA. The polyanions heparin and chondroitin sulfates A and B do not inhibit binding. These observations suggest the involvement of specific receptor proteins in binding of oligomers to mammalian cells. PMID- 2549539 TI - Mutually exclusive use of viral promoters in Epstein-Barr virus latently infected lymphocytes. AB - Of the eight viral antigens known to be expressed during Epstein-Barr virus latency, six are transcribed from a major rightward transcriptional unit, which gives rise to mRNAs containing common 5' exons. Analysis of cDNA clones has identified the use of two different promoters (Wp and Cp), located near the left hand end of the viral genome, in generating these viral messages. Characterization of the activities of these two viral promoters in a number of Burkitt lymphoma and lymphoblastoid cell lines has revealed exclusive usage of only one of these promoters in all cell lines examined. Transfection of reporter constructs containing Wp and/or Cp linked to the bacterial chloramphenicol acetyltransferase gene into several different Epstein-Barr virus-infected cell lines generally supports a model in which the mutually exclusive use of Cp or Wp is determined by cellular factors and not by viral strain variation. PMID- 2549538 TI - Production of simian virus 40 large tumor antigen in bacteria: altered DNA binding specificity and dna-replication activity of underphosphorylated large tumor antigen. AB - A bacterial expression system was used to produce simian virus 40 large tumor antigen (T antigen) in the absence of the extensive posttranslational modifications that occur in mammalian cells. Wild-type T antigen produced in bacteria retained a specific subset of the biochemical activities displayed by its mammalian counterpart. Escherichia coli T antigen functioned as a helicase and bound to DNA fragments containing either site I or the wild-type origin of replication in a manner identical to mammalian T antigen. However, T antigen purified from E. coli did not efficiently bind to site II, an essential cis element within the simian virus 40 origin of replication. It therefore could not unwind origin-containing plasmids or efficiently replicate simian virus 40 DNA in vitro. The ability of protein phosphorylation to modulate the intrinsic preference of full-length T antigen for either site I or site II is discussed. PMID- 2549540 TI - Cellular proteins specifically bind single- and double-stranded DNA and RNA from the initiation site of a transcript that crosses the origin of DNA replication of herpes simplex virus 1. AB - The small-component origins of herpes simplex virus 1 DNA synthesis are transcribed late in infection by an RNA with heterogeneous initiation sites approximately 290-360 base pairs from the origins. We report that cellular proteins react with a labeled RNA probe representing the 5' terminus of a subset of this RNA but not with the complementary strand of this RNA. The proteins form two complexes. Complex 2 was formed by all nuclear extracts tested, whereas complex 1 was invariably formed by proteins present only in nuclear extracts of mock-infected cells. Complex 1 protects a contiguous stretch of 40 nucleotides of the labeled RNA probe from nuclease degradation. Formation of complex 1 was competitively inhibited in a sequence-specific fashion by single-stranded RNA and DNA and by double-stranded RNA and DNA. The protein(s) forming complex 1 is, thus, quite distinct from known nucleic acid-binding proteins in that they recognize a specific nucleotide sequence, irrespective of the nature (single- and double-stranded RNA and DNA) of the nucleic acid. We conclude the following: (i) the proteins forming complex 1 and 2 are probably different, (ii) complex 1 is neither required throughout infection for viral replication nor able to hinder viral replication in cells in culture, and (iii) cells susceptible to infection encode one or more proteins that recognize specific sequences in single-stranded nucleic acids; either these proteins impart a compatible conformation on single stranded nucleic acids with the conformation of the same strand in the double stranded nucleic acid, or these proteins confer a specific, distinct conformation to both single-stranded and double-stranded nucleic acids. PMID- 2549541 TI - 5' flanking sequence and structure of a gene encoding rat 6-phosphofructo-2 kinase/fructose-2,6-bisphosphatase. AB - The synthesis and degradation of fructose 2,6-bisphosphate, a ubiquitous stimulator of glycolysis, are catalyzed by 6-phosphofructo-2-kinase (EC 2.7.1.105) and fructose-2,6-bisphosphatase (EC 3.1.3.46), respectively. In liver, these two activities belong to separate domains of the same 470-residue polypeptide. Various mRNAs have been described for this bifunctional enzyme, which is controlled by hormonal and metabolic signals. To understand the origin and regulation of these mRNAs, we have characterized rat genomic clones encoding the liver isozyme, which is regulated by cAMP-dependent protein kinase, and the muscle isozyme, which is not. We describe here a 55-kilobase gene that encodes these isozymes by alternative splicing from two promoters. Each of the putative promoters was sequenced over about 3 kilobases and found to include nucleotide motifs for binding regulatory factors. The two isozymes share the same 13 exons and differ only by the first exon that, in the liver but not in the muscle isozyme, contains the serine phosphorylated by cAMP-dependent protein kinase. The gene was assigned to the X chromosome. An analysis of the exon limits of 6 phosphofructo-2-kinase/fructose-2,6-bisphosphatase in relation to its functional domains and to its similarity with other proteins plus its G + C content at the third codon position suggests that this gene originates from several fusion events. PMID- 2549542 TI - Sequences required for expression of Bordetella pertussis virulence factors share homology with prokaryotic signal transduction proteins. AB - The bvg locus of Bordetella pertussis is required for coordinate regulation of several factors associated with virulence. The control system is modulated by various environmental signals, including low temperature, MgSO4, and nicotinic acid. The nucleotide sequence of the bvg region has been determined and three open reading frames, bvgA, bvgB, and bvgC, are present. Twelve-base-pair linker insertion mutations in any of these open reading frames result in a Bvg- phenotype. The predicted protein products of bvgA and bvgC share homology with a family of prokaryotic regulatory proteins that respond to environmental stimuli and are members of two-component sensory transduction systems. We propose a model in which BvgB and the N-terminal portion of BvgC are localized in the periplasm. Environmental signals are recognized, transduced to the cytoplasmic portion of BvgC, and then transmitted to BvgA, a positive regulator of transcription. PMID- 2549543 TI - Genome organization of the anaerobic pathogen Clostridium perfringens. AB - A physical map of the genome of Clostridium perfringens, an important human pathogen, has been established by pulsed-field gel electrophoresis. Recognition sites for six rare-cutting endonucleases were situated on a single circular chromosome of approximately 3.6 million base pairs thus defining 50 arbitrary genetic intervals of between 10 and 250 kilobase pairs. This considerably facilitated the chromosomal localization of some 24 genes and loci for which probes were available and allowed the construction of the genome map of a clostridial species. PMID- 2549544 TI - Surface immunoglobulin-mediated B-cell activation in the absence of detectable elevations in intracellular ionized calcium: a model for T-cell-independent B cell activation. AB - We recently showed that anti-immunoglobulin conjugated to high molecular weight dextran is 1000-fold more mitogenic for B cells than unconjugated anti immunoglobulin. This system serves as a model for T-cell-independent type 2 antigens such as haptenated Ficoll, dextran, and bacterial polysaccharides, which can also stimulate B-cell proliferation and antibody production at low concentrations. We show here that conjugated anti-immunoglobulin, at concentrations that stimulate significant increases in expression of major histocompatibility complex class II molecules and incorporation of thymidine into DNA, does not induce detectable modulation of surface immunoglobulin. These results indicate that the facilitated T-cell-independent B-cell activation by polysaccharide antigens may result from inability to modulate surface immunoglobulin, possibly resulting in persistent and/or repetitive signaling. Early large increases in Ca2+ and breakdown of inositol phospholipids presently thought to be involved in transduction of the mitogenic signal are not detectable at low concentrations of conjugated anti-immunoglobulin, raising the possibility that these biochemical events may not in fact be central to this signaling pathway. PMID- 2549545 TI - Retroviral vector-mediated high-efficiency expression of adenosine deaminase (ADA) in hematopoietic long-term cultures of ADA-deficient marrow cells. AB - Two recombinant retroviral vectors encoding the cDNA of the human adenosine deaminase (ADA; EC 3.5.4.4) gene and the bacterial neomycin resistance (Neo) gene have been used to transduce bone marrow cells obtained from four patients affected by the ADA-deficient variant of severe combined immunodeficiency. By utilizing the long-term marrow culture system, freshly isolated bone marrow cells were subjected to multiple infection cycles with cell-free supernatants containing high titers of viral vector and then maintained in long-term marrow culture in the absence of any overt selection pressure. By using this experimental protocol, about 30-40% of the hematopoietic progenitors were productively transduced with the viral vector, as judged by the appearance of G418-resistant colonies derived from granulocyte/macrophage and multipotent hematopoietic progenitor cells. The vector-encoded human ADA gene was expressed efficiently in both the myeloid and lymphoid progeny of the cultured bone marrow cells, reaching levels between 15% and 100% as compared to the levels of ADA in normal bone marrow cells. The efficiency of gene transfer and ADA production was proportional to the number of infection cycles. Furthermore, transduction of the ADA vectors into the bone marrow cells derived from an ADA-deficient patient restored the capacity of the cells to respond to phytohemagglutinin and interleukin 2. PMID- 2549546 TI - Interleukin 1 and tumor necrosis factor inhibit cardiac myocyte beta-adrenergic responsiveness. AB - Reversible congestive heart failure can accompany cardiac allograft rejection and inflammatory myocarditis, conditions associated with an immune cell infiltrate of the myocardium. To determine whether immune cell secretory products alter cardiac muscle metabolism without cytotoxicity, we cultured cardiac myocytes in the presence of culture supernatants from activated immune cells. We observed that these culture supernatants inhibit beta-adrenergic agonist-mediated increases in cultured cardiac myocyte contractility and intracellular cAMP accumulation. The myocyte contractile response to increased extracellular Ca2+ concentration is unaltered by prior exposure to these culture supernatants, as is the increase in myocyte intracellular cAMP concentration in response to stimulation with forskolin, a direct adenyl cyclase activator. Inhibition occurs in the absence of alteration in beta-adrenergic receptor density or ligand binding affinity. Suppressive activity is attributable to the macrophage-derived cytokines interleukin 1 and tumor necrosis factor. Thus, these observations describe a role for defined cytokines in regulating the hormonal responsiveness and function of contractile cells. The effects of interleukin 1 and tumor necrosis factor on intracellular cAMP accumulation may be a model for immune modulation of other cellular functions dependent upon cyclic nucleotide metabolism. The uncoupling of agonist-occupied receptors from adenyl cyclase suggests that beta-receptor or guanine nucleotide binding protein function is altered by the direct or indirect action of cytokines on cardiac muscle cells. PMID- 2549547 TI - Localized actions of progesterone in hypothalamus involve oxytocin. AB - Two ovarian hormones, estradiol and progesterone, which facilitate mating behavior in the female rat by acting on the ventromedial nuclei (VMN) of the hypothalamus, induce changes in oxytocin receptor binding in this brain region. Estradiol induced a 4-fold increase in the oxytocin receptor binding of the VMN and surrounding area and increased the number and immunostaining of oxytocin fibers in an area lateral to the ventral VMN. Progesterone, in estrogen-primed rats, caused the induced oxytocin receptors to spread over the area containing the oxytocin fibers. Infusion of oxytocin into the ventromedial hypothalamus increased the display of lordosis behavior only in females primed with both estradiol benzoate and progesterone. Thus, the sequential actions of two ovarian hormones bring a neuropeptide and its receptors into register and enable the neuropeptide to exert behavioral effects. PMID- 2549548 TI - Transient low-threshold Ca2+ current triggers burst firing through an afterdepolarizing potential in an adult mammalian neuron. AB - In a variety of mammalian neurons, a brief depolarization generates an afterdepolarizing potential that triggers the firing of a short series or burst of action potentials. Although such burst firing is thought to contribute to the processing of neural information, the ionic currents that underlie this phenomenon have not been established. In whole-cell patch-clamp experiments on dorsal root ganglion neurons, we have found that the current that underlies this type of burst firing is a transient low-threshold (T-type) Ca2+ current. The data suggest that the T-type Ca2+ current may play an important role in the processing of information in the nervous system by virtue of its ability to elicit burst firing in neurons. PMID- 2549549 TI - cGMP induces phase shifts of a mammalian circadian pacemaker at night, in antiphase to cAMP effects. AB - The suprachiasmatic nuclei (SCN) of mammals contain a circadian clock that synchronizes behavioral and physiological rhythms to the daily cycle of light and darkness. We have been probing the biochemical substrates of this endogenous pacemaker by examining the ability of treatments affecting cyclic nucleotide dependent pathways to induce changes in the phase of oscillation in electrical activity of rat SCN isolated in brain slices. Our previous work has shown that daytime treatments that stimulate cAMP-dependent pathways induce phase shifts of the SCN pacemaker in vitro but treatments during the subjective night are without effect. In this study we report that the phase of SCN oscillation is reset by treatments that stimulate cGMP-dependent pathways, but only during the subjective night. Thus, the nocturnal period of SCN sensitivity to cGMP is in antiphase to the diurnal period of sensitivity to cAMP. These results suggest that cAMP and cGMP affect the SCN pacemaker through separate biochemical pathways intrinsic to the SCN. These studies provide evidence that changing biochemical substrates within the SCN circadian clock may underlie some aspects of differential temporal sensitivity of mammals to resetting stimuli. PMID- 2549551 TI - Voltage-activated K+ conductances in freshly isolated embryonic chicken osteoclasts. AB - Patch-clamp measurements on freshly isolated embryonic chicken osteoclasts revealed three distinct types of voltage-dependent K+ conductance. The first type of conductance, present in 72% of the cells, activated at membrane potentials less negative than -30 to -20 mV and reached full activation at +40 mV. It activated with a delay, reached a peak value, and then inactivated with a time constant of approximately 1.5 s. Inactivation was complete or almost so. Recovery from inactivation, at -70 mV, had a time constant of roughly 1 s. The conductance could be blocked, at least partly, by 4 mM 4-aminopyridine. The second type of conductance (present in all cells) activated at membrane potentials more negative than -40 to -80 mV and reached full activation at -130 mV. Activation potential and maximal conductance were dependent on the extracellular K+ concentration. Inactivation of the conductance first became apparent at membrane potentials more negative than -100 mV and was a two-exponential process. The conductance could be blocked by external 5 mM Cs+ ions. The third type of conductance (present in all cells) activated at membrane potentials more positive than +30 mV. Generally, the conductance did not inactivate. PMID- 2549550 TI - Activation of purified calcium channels by stoichiometric protein phosphorylation. AB - Purified dihydropyridine-sensitive calcium channels from rabbit skeletal muscle were reconstituted into phosphatidylcholine vesicles to evaluate the effect of phosphorylation by cyclic AMP-dependent protein kinase (PK-A) on their function. Both the rate and extent of 45Ca2+ uptake into vesicles containing reconstituted calcium channels were increased severalfold after incubation with ATP and PK-A. The degree of stimulation of 45Ca2+ uptake was linearly proportional to the extent of phosphorylation of the alpha 1 and beta subunits of the calcium channel up to a stoichiometry of approximately 1 mol of phosphate incorporated into each subunit. The calcium channels activated by phosphorylation were determined to be incorporated into the reconstituted vesicles in the inside-out orientation and were completely inhibited by low concentrations of dihydropyridines, phenylalkylamines, Cd2+, Ni2+, and Mg2+. The results demonstrate a direct relationship between PK-A-catalyzed phosphorylation of the alpha 1 and beta subunits of the purified calcium channel and activation of the ion conductance activity of the dihydropyridine-sensitive calcium channels. PMID- 2549553 TI - Heterogeneity within an HSV-1 wild-type strain and its importance in pathogenesis. AB - A herpes simplex virus-type 1 low passage, clinical eye isolate, E-43 at P2, was compared with its variant progeny, SLi-43 at P8, in terms of ocular disease, cytopathic effects, and genomic variation. In New Zealand White (NZW) rabbits, E 43 produced mild epithelial defects and punctate lesions with full recovery by Day 10 postinfection (pi). SLi-43 caused dendritic lesions, progressing to geographic ulceration and death from herpes simplex virus encephalitis in 10 days postinfection. In RK, Hep-2, and Vero cells, E-43 displayed the syn+ phenotype (aggregation and cell rounding); SLi-43 showed the syn phenotype (syncytium formation). DNA digestion profiles of E-43, SLi-43, and isolates from the brains of infected animals showed that the genomic differences map within the terminal repeat of the unique long segment and the internal joint region, specifically in bands B, E, N, and S (Bam HI) and bands M and N (Hind III). Analysis of the DNA of virus recovered from the brain stem of SLi-43-infected, encephalitic rabbits demonstrated that an in vivo selection for neurotropic virions had taken place. Plaque purification of 20 clones from the original E-43 strain showed that one of 20 was the syn phenotype, indicating that the SLi-43 variant was present in the original E-43 isolate and did not develop de novo by rapid mutation. The parent progeny relationship between E-43 and SLi-43 forms an ideal model in which to compare differences in pathogenicity at the genomic level, and underscores the importance of heterogeneity within a single herpes simplex virus-type 1 wild-type population in terms of variations in ocular disease. PMID- 2549552 TI - Parathyroid hormone and parathyroid hormone-related protein inhibit phasic contraction of pig duodenal smooth muscle. AB - Parathyroid hormone (PTH) and a newly discovered PTH-related protein (PTHrP), which has amino-terminal homology with PTH, are potent relaxants of rat gastrointestinal tissues. Since their gastrointestinal relaxant effects have been described only in the rat, we examined their actions in another mammalian species in order to evaluate whether the relaxant property was more generally applicable. Longitudinal smooth muscle strips were obtained from the pig duodenum. The mucosa was removed, the strips were mounted in a tissue chamber, and changes in phasic contraction were detected with a force-displacement transducer and recorded using a polygraph. Acetylcholine-induced phasic contraction was inhibited rapidly in a dose-related manner by [Nle8,18,Tyr34]-bPTH-(1-34)-amide, or hPTHrP-(1-34). The IC50 values for these peptides were 2.6 nM and 6.1 nM, respectively. The maximal effect of both peptides was observed at 60 nM with an 84% decrease of the acetylcholine-induced contraction. At 400 nM, the PTH antagonist, [Nle8,18,Tyr34] bPTH-(3-34)-amide, had no effect by itself. However, the same 400 nM concentration of this peptide totally blocked the decrease in phasic contraction induced by 10 nM of the bPTH-(1-34) analogue or hPTHrP-(1-34). Our results show that receptors for PTH or PTHrP are present in the muscular layer of the pig duodenum and that activation of these receptors inhibits the phasic contraction of the tissue. Furthermore, the ability of PTH-related peptides to relax gastrointestinal smooth muscle is not restricted to the rat. PMID- 2549554 TI - Activation of human neutrophils and oxygen radical scavenging activity of phenyl 3-methoxy-4-hydroxy styryl ketone. PMID- 2549555 TI - [Animal experiment studies of the effect of the hazardous substance carbon tetrachloride on peripheral nerves]. AB - The deleterious effects of Carbon Tetrachloride, widely used industrially, on the peripheral nervous system was investigated by experimentation on rabbits. Long term infusion simulated various conditions of exposure to obtain pharmacological and electroneurographic data. Tetrachlorethen is shown to induce reversible dysfunctions of the peripheral nervous system depending on the degree of concentration. The mode of molecular action remains open. Our results are compared with references in existing literature. PMID- 2549556 TI - 3H-imipramine binding to freshly prepared platelet membranes in depression. AB - 3H-Imipramine binding was measured in freshly prepared platelet membranes from 47 drug-free major depressives and 46 healthy controls. Where possible, platelet binding in depressed subjects was repeated following treatment. A significant negative correlation was found between Bmax and assay protein concentration and Bmax values were corrected for this effect. Adjusted Bmax was significantly lower (by 14%) in female depressed patients than in female control subjects, and the difference was of similar magnitude premenopausally and postmenopausally. No such difference was found in males. Kd did not differ significantly between depressed and control subjects. Multiple regression analysis confirmed significant effects on Bmax of presence of depressive illness, age (positive correlation), and season (higher in summer). Within the depressed sample, Bmax was significantly lower in those subjects with obsessional features. Endogenicity (Research Diagnostic Criteria or Newcastle), dexamethasone suppression test result, drug-free interval, family history of depression, depressive psychosis, suicidal ideation, and past history of suicide attempts were not significantly related to Bmax. Paired comparisons revealed no significant effect on Bmax of 6 weeks' treatment with imipramine, maprotiline, or BRL 14342 or of a course of electroconvulsive therapy. PMID- 2549557 TI - Naltrexone potentiates glycemic responses during stress and epinephrine challenge in genetically obese mice. AB - The genetically obese mouse (C57BL/6J ob/ob) is a commonly used animal model of non-insulin-dependent diabetes mellitus. These mice show exaggerated glycemic responses during behavioral stress and adrenergic stimulation, but the precise glucoregulatory mechanisms are not well characterized. The ob/ob mice have multiple endocrine abnormalities, including elevated pituitary and circulating beta-endorphin levels; and a relationship between hyperglycemia and altered opioid function has been suspected. We now report that opiate antagonism with naltrexone potentiates hyperglycemic responses during stress and epinephrine challenge in obese mice. This effect of opioid blockade suggests that endogenous opioids inhibit stress- and epinephrine-induced hyperglycemia in the genetically obese mouse. PMID- 2549558 TI - The effect of propranolol on behavioral interactions among adult male cynomolgus monkeys (Macaca fascicularis) housed in disrupted social groupings. AB - We report here on the social behavior of 30 adult male cynomolgus macaques (Macaca fascicularis), half of which were chronically administered propranolol HCl. All monkeys were maintained in groups of five, fed an atherogenic diet, and subjected to a social manipulation involving repeated disruption of social groups through redistribution of group memberships. One year of intensive behavioral observations were made, involving quantification of 27 kinds of social interaction typically exhibited by this species in captivity. Propranolol treatment resulted in a significant reduction in heart rate and blood pressure. Comparisons between propranolol-treated and untreated monkeys revealed no effects of beta-blockade on social dominance, aggressiveness, or active aspects of affiliation (e.g., grooming). The fact that few behavioral differences were found between conditions in this investigation indicates to us that such effects as were observed may owe as much to chance as to the action of the drug. Overall, these findings suggest that chronic beta-adrenergic blockade has minimal effects on the most prominent features of macaque monkey social behavior, especially those behavioral factors (aggressiveness and competitiveness) associated with risk for coronary disease in this species and in human beings. PMID- 2549559 TI - The seroprevalence of cytomegalovirus among Virginia State prisoners. AB - A prevalence serosurvey of adult male prisoners entering the Virginia State Prison was conducted to evaluate the epidemiology of cytomegalovirus within this population. Four hundred and forty-five (97%) of 459 male inmates provided serum for analysis and 427 completed a detailed demographic questionnaire. Sera were tested for cytomegalovirus by passive latex agglutination and 64% were reactive. Multivariate discriminant analysis showed an independent association of seropositivity with age, non-white race, and a history of gonorrhea. There was no apparent contribution from admitted homosexual contact though this may have been under-reported. There was no correlation of seropositivity with intravenous drug use or with the length or number of prior incarcerations. Prisoners possess the same correlates for cytomegalovirus seropositivity as the general adult population; past imprisonment did not independently contribute to cytomegalovirus seropositivity. PMID- 2549560 TI - Measurement of radiation-induced DNA damage using gel electrophoresis or neutral filter elution shows an increased frequency of DNA strand breaks after exposure to pH 9.6. AB - The filter elution technique using nondenaturing conditions is widely used to assay DNA double-strand break (DSB) induction and repair. It has been reported that in the measurement of strand breaks higher rates of elution and of initial rejoining are obtained at pH 9.6 compared to pH 7.2. In the present experiments neutral elution at pH 7.2 and 9.6 were compared in the assay of damage to DNA induced by X rays, 125I decay, and restriction enzyme digestion, in an effort to explain this discrepancy and to determine whether the higher rate of elution observed at pH 9.6 corresponds to a greater number of DSBs. X-ray damage to cellular DNA resulted in significantly different elution profiles at the two pH values. In contrast the elution profiles of the DSB induced by intragenomic 125I decays or restriction endonuclease were independent of the pH of the elution buffer. When gamma-irradiated SV40 DNA was exposed to pH 7.2 or 9.6 elution buffer prior to analysis by gel electrophoresis, a significantly greater number of DNA DSBs were detected in the DNA exposed to pH 9.6. We conclude that X and gamma radiation produce lesions (pH 9.6-labile lesions), in proportion to dose, that have the potential of becoming measurable DSBs following incubation under the mildly alkaline condition of pH 9.6. The data suggest that these lesions may result from single-hit events. PMID- 2549561 TI - Electrophoretic mobility of erythrocytes from adeturone-pretreated rats exposed to gamma or high-energy protons. AB - Change in electrophoretic mobility (EM) of erythrocytes was studied 6 hours after rats were exposed to a dose range (0.5-4 Gy) of high-energy protons (9 GeV/nucleon) or gamma rays (0.660 MeV), with or without adeturone pretreatment (300 mg/kg body weight). With either of the two radiation types used, EM of the cells was observed to undergo significant dose-related decline. As indicated by our findings, adeturone pretreatment failed to contribute to maintenance of normal erythrocyte surface charge in the case of proton irradiation, proving, nevertheless, effective against gamma-ray exposure at all dose levels used. PMID- 2549562 TI - Radon: is it a problem? AB - Radon gas is a major source of radiation exposure to the general public. Radon 222 is a product of uranium-238, present in varying concentrations in all soils. Radon enters buildings from soil, water, natural gas, and building materials. Its short-lived breakdown products, termed "radon daughters," include alpha-emitting solids that can deposit in the lungs. Firm evidence links lung cancer risk in miners with high exposure to radon daughters. The amount of risk associated with the much lower but chronic doses received in buildings is difficult to establish. By some extrapolations, radon daughters may be responsible for a significant number of lung cancer deaths. The existence or extent of synergism with smoking is unresolved. Local conditions can cause high levels of radon in some buildings, and measures that reduce indoor radon are of potential value. PMID- 2549563 TI - Ductal carcinoma in situ: atypical mammographic appearances. AB - The authors retrospectively analyzed the mammograms of 190 women with biopsy proved ductal carcinoma in situ (DCIS). Excluded from the current study were 117 (62%) women whose radiographs showed suspicious clustered microcalcifications, a well-known finding in DCIS. Of the remaining 73 (38%) women, 30 (16%) had negative mammograms, and 43 (23%) had mammographic manifestations of breast malignancy other than microcalcifications. Of the latter 43, 15 had circumscribed masses, and 12 had various focal nodular patterns. The remaining 16 patients showed other mammographic signs of malignancy, including asymmetry (n = 1); dilated retroareolar ducts (n = 2); ill-defined, rounded tumor (n = 2); focal architectural distortion (n = 4); subareolar mass (n = 3); and developing density (n = 4). Of the 73 women in the study, 60 presented with clinical findings related to the tumor. Since DCIS has a high survival rate with proper treatment, radiologists should be aware of the unusual radiographic manifestations of this disease. PMID- 2549564 TI - Fibroadenoma of the breast: sonographic appearance. AB - The sonographic patterns of 100 fibroadenomas, including two giant fibroadenomas and two cystosarcoma phyllodes, were reviewed. The lesions were mostly hypoechoic (92%) and homogeneous in echotexture (72%). Twenty-seven percent of fibroadenomas showed irregular margins. Intratumoral calcifications were noted in 10% of lesions. A marked acoustic shadow without associated calcification was seen in six cases. The mean ratio of the length to the anteroposterior diameter of the tumors was 1.84 +/- 0.52 (standard deviation), indicating an elongation along the general orientation of the breast tissue planes. The ratios of length to anteroposterior diameter calculated in two matching groups of small (less than 1 cm3) fibroadenomas and carcinomas differed significantly. State-of-the-art high frequency transducers, geometric analysis of tumors, and real-time ultrasound guided fine-needle aspiration biopsy should help to distinguish between benign and malignant solid breast masses. PMID- 2549565 TI - Hilar cholangiocarcinoma: comparative study with sonography and CT. AB - Fifty-one patients were studied to determine the relative accuracy of sonography and computed tomography (CT) in the evaluation of hilar cholangiocarcinoma. Both sonography and CT permitted detection of intrahepatic bile duct dilatation in all patients. Nonunion of right and left intrahepatic bile ducts was identified with sonography in 90% and with CT in 94% of patients. The tumor per se was depicted by sonography in 21% and by CT in 40% of patients. Forty tumors (78%) were of the infiltrating type, seven (14%) were exophytic, and four (8%) were polypoid. With sonography, infiltrating and exophytic tumors were difficult to depict, whereas polypoid tumors were well identified. With CT, infiltrating tumors were more difficult to depict than were exophytic or polypoid tumors. On CT scans, depicted tumors of the infiltrating type showed high attenuation and an indistinct margin, whereas exophytic and polypoid tumors showed low attenuation and a well-defined margin. Sonography and CT were comparably accurate in determining the level of obstruction in hilar cholangiocarcinoma even when no mass was depicted. CT was superior to sonography in depicting tumor per se and in demonstrating associated findings. PMID- 2549566 TI - Unresectable non-oat cell carcinoma of the lung: definitive radiation therapy. AB - Between 1976 and 1983, 267 patients with non-oat cell carcinoma of the lung were treated with radiation therapy alone. One hundred thirty-four patients had squamous cell carcinoma; 69, large cell carcinoma; and 64, adenocarcinoma. Stage III carcinoma was diagnosed in 87% of the patients. Total radiation dose was less than 45 Gy in 69 patients (low dose group), 45-55 Gy in 161 (middle dose group), and 55-65 Gy in 37 (high dose group); dosage was 180-200 cGy daily, 5 days per week. Minimum follow-up was 3 years (median, 6 years). Tumor control within the radiation fields was achieved in 12%, 43%, and 78% of the low, middle, and high dose groups, respectively. A complete response rate of 13%, 23%, and 35% and an overall response of 43%, 71%, and 86% were seen in the low, middle, and high dose groups, respectively. The 5-year recurrence-free survival rate for all patients was 7% and was dependent on radiation dose and tumor response. This study indicates that tumor control and complete response rates are improved with a radiation dose of 55-65 Gy and that complete responders have improved survival. PMID- 2549567 TI - Hepatocellular carcinoma: pilot trial of treatment with Y-90 microspheres. AB - The potential use of yttrium-90 glass microspheres in the treatment of hepatocellular carcinoma was assessed in a pilot study of seven patients. The Y 90 microspheres were injected via a hepatic artery catheter. In this group of patients, no toxicity was observed for absorbed doses of between 5,000 and 10,000 cGy to the liver and up to 32,000 cGy to the tumor itself. Tumor response was seen only at the higher absorbed doses. The new Y-90 glass microspheres can safely deliver large doses of internal radiation to hepatic tumors as long as extrahepatic shunting can be excluded. Extrahepatic shunting will be the main limitation to this form of radiation therapy. PMID- 2549568 TI - [HPV infections in tumors of the upper respiratory and digestive tract. In situ hybridization and dot-blot hybridization with biotinylated DNA probes]. PMID- 2549569 TI - [Detection of viral nucleic acid sequences in paraffin and cryostat sections of clinical study samples using in situ hybridization]. PMID- 2549570 TI - [In situ hybridization in detecting virus infections in biopsies following orthotopic liver transplantation]. AB - Using the in-situ DNA hybridization we studied 62 liver biopsies from 34 patients for the presence of CMV-, EBV-, HBV-, and HSV-Genome. In the first biopsies positivity-rates were as follows: CMV 62%, EBV 71%, HBV 15%, HSV 15%. In 6 cases a conversion from a negative intranuclear CMV-status to a positive status was seen correlating with an increase in serological titers and with clinical signs of viral infection. PMID- 2549571 TI - Regionality of glucose-6-phosphate hydrolysis in the liver lobule of the rat. Metabolic heterogeneity of 'portal' and 'septal' sinusoids. PMID- 2549572 TI - [Detection of HPV 6/11 and 16/18 in normal cervix epithelia and in condylomatous lesions. Comparative study of in situ hybridization and dot-blot hybridization using the streptavidin-alkaline phosphatase system]. PMID- 2549573 TI - Metabolism and excretion of exogenous [3H]-LTC4 in primates. AB - Four novel omega- and beta-oxidation (from the omega end) products of peptide leukotrienes, 20-hydroxy and 20-carboxy-LTE4, 18-carboxy-19, 20-dinor-LTE4 and 16 carboxy-17,18,19,20-tetranor-14,15-dihydro-LTE4 were prepared by total synthesis and used as standards for identification of biliary and urinary metabolites in the cynomolgus monkey. After intravenous administration 14, 15-[3H] leukotriene C4 (10 microCi kg-1) was partially metabolized in and rapidly cleared from the vascular circulation. This resulted, within 24 hours, in significant urinary excretion (14.8 +/- 2.1%, n = 4), consisting largely of material more polar than LTE4 (61% of urinary excretion) as shown by reverse phase HPLC. The polar fraction demonstrated two predominant metabolites which coeluted in several HPLC solvent systems with synthetic 16-carboxytetranordihydro-LTE4 (major component) and 18-carboxydinor-LTE4 (minor component). Characterization of the major polar metabolite as 16-carboxytetranordihydro-LTE4 was substantiated by conversion to its N-acetylated derivative. The absence of the 14, 15 double bond was confirmed by product analysis of oxidative ozonolysis. In a single animal, the bile duct was cannulated, with significant biliary excretion of radioactivity demonstrated over 4 hours (58.6% recovery). The predominant polar biliary metabolites were also identified as the 18-carboxydinor and 16-carboxytetranordihydro derivatives of LTE4 mentioned above. These data suggest that beta-oxidation products generated from the omega-carboxyl end of the 20-carboxy-LTE4 are important products of [3H] LTC4 metabolism in the monkey. Quantitation of these urinary metabolites may be an important index of in vivo leukotriene production. PMID- 2549574 TI - Prostaglandin binding sites in human polymorphonuclear neutrophils. AB - Prostaglandin (PG) E2 (greater than or equal to 1.6 nM) and PGD2 (greater than or equal to 16 nM) inhibited polymorphonuclear neutrophil (PMN) degranulation responses to leukotriene (LT) B4 and platelet-activating factor (PAF) whereas PGF2 alpha was bioinactive. [3H]PGE2 and [3H]PGD2 bound to PMN and isolated, plasmalemma-enriched PMN membranes. Binding was time-dependent, specific, saturable, and reversible. Competitive studies indicated that the two PGs bound to distinctly different sites. PMN had high (Kd = 1 nM; Rt = 150/cell) and low (Kd = 100 nM; Rt = 5800/cell) affinity PGE2 binding sites. Only a single type of PGD2 binding site (Kd = 13 nM; Rt = 5100/cell) was detected. We conclude that PGE2 and PGD2 bind to their respective, plasmalemmal receptors to attenuate PMN function. The PGs may act as endogenous stop signals to limit the action of concurrently formed excitatory signals, eg., LTB4 and PAF. PMID- 2549575 TI - Prostaglandins and growth of the adrenal cortex. PMID- 2549576 TI - Ovarian follicular fluid eicosanoid concentrations during the pre-ovulatory period in humans. AB - Prostaglandins are involved in ovulation and in every mammal studied so far, ovulation has been inhibited by prostaglandin inhibition. Information regarding the role of leukotrienes and thromboxanes in ovulation is more limited. In order to study the production of eicosanoids in human pre-ovulatory follicular fluid, follicular aspiration was timed by means of serial ultrasound scans and human chorionic gonadotrophin (hCG) to be immediately pre-ovulatory. 11 women were studied and the eicosanoids measured by radioimmunoassay (RIA). The follicular fluid was found to contain leukotrienes (LT) B4, LTC4 (D4, E4), prostaglandin (PG) E2, PGF2 alpha 6 keto PGF1 alpha k and thromboxane (TX) B2. This is the first published report of leukotrienes in human follicular fluid in spontaneous cycles, and is one of the few reports showing prostaglandins and thromboxanes. The significance of demonstrating leukotrienes in human follicular fluid is discussed as is the correlation between individual eicosanoids in the human ovary. PMID- 2549577 TI - Metabolism of leukotriene D4 by rat peritoneal mast cells. AB - Metabolism of sulfidopeptide leukotrienes, leukotrienes (LT) C4 and D4 by rat peritoneal mast cells was studied. Rat peritoneal mast cells converted LTD4 to LTE4 but not LTC4 to LTD4. The LTD4-metabolizing activity was equally distributed on the cell surface and inside cells, but not released to the extracellular milieu even when a considerable portion of histamine and the secretory granule enzymes were released. Among various enzyme inhibitors examined, o phenanthroline, a metal chelator, and dithiothreitol significantly suppressed the LTD4-metabolizing activity of mast cell. These results would suggest that some metalloenzyme located on the cell surface is involved in the conversion of LTD4 to LTE4 by rat peritoneal mast cells. PMID- 2549579 TI - [Regulating interpretation of the mechanisms of occurrence of epileptic activity in neuronal networks. III. Synchronization, continuous epileptic activity and possible epileptic destabilization of the central nervous system]. PMID- 2549578 TI - Dietary marine lipid does not alter susceptibility of (NZBxNZW)F1 mice to pathogenic microorganisms. AB - Female (NZBxNZW)F1 mice were fed for one month with a diet in which the sources of fat were either melted beef tallow or fish oil, the latter regimen being associated with a marked decrease in the expression of auto-immune disease in these animals. To test whether or not this beneficial effect was associated with an increased risk of infection, animals fed each of these diets were challenged intraperitoneally with graded doses of four different classes of microorganisms: a gram positive bacillus that is an intracellular parasite (Listeria monocytogenes), an exotoxin-producing gram negative bacillus (Pseudomonas aeruginosa), a yeast (Candida albicans), and a herpes group virus (murine cytomegalovirus). There was no difference in the susceptibility of the animals fed the two different diets to any of these infections as shown by either determinations of the LD50 for each organism or by assessment of the times of survival for those animals that did succumb. We conclude that the beneficial anti inflammatory effects of fish oil diets in these mice are not associated with an increased risk of infection with a variety of microbial agents. PMID- 2549580 TI - [Enalapril in the treatment of primary arterial hypertension]. AB - Enalapril (20 mg daily) was used in the treatment of primary arterial hypertension in 28 patients with the WHO class I and II and diastolic blood pressure ranging from 105 to 120 mm Hg. Hypotensive effect after an initial dose of enalapril occurred in 12 patients (42.8%) in whom a decrease in blood pressure at 1 hr exceeded 10% of the basal value (from means = 169.9/109.1 to 146.7/97.9 mmHg). At 6 weeks positive hypotensive effect was found in 46% of patients. A combination of enalapril with hydrochlorothiazide (25 mg daily) in patients without normalization of the blood pressure level increased the number of the favourably treated to 64%. Of 12 patients with positive effects after the first dose of enalapril only seven patients profited from chronic enalapril monotherapy. In chronic treatment enalapril seemed to be well tolerated and side effects were only sporadic. PMID- 2549582 TI - Prognostic factors for patients with inoperable non-small cell lung cancer, limited disease. The importance of patients' subjective experience of disease and psychosocial well-being. AB - In a prospective controlled clinical trial, 102 patients with inoperable non small cell lung cancer (NSCLC), limited disease, stage II and III were treated with combination chemotherapy, cisplatin 70 mg/m2 i.v. on day one and etoposide 100 mg/m2 i.v. on day one, and etoposide 200 mg/m2 orally on days 2 and 3, or radiotherapy given in 15 fractions of 2.8 Gy with two anterior/posterior fields during a period of three weeks. The patients completed a validated self administered questionnaire before the start of treatment that assessed their psychosocial well-being, disease-related symptoms, personal functioning, and everyday activity. These subjective variables were evaluated together with treatment modality, WHO performance status, weight loss, and stage of disease, with regard to their value in predicting survival. Univariate survival analyses were undertaken for each individual factor, median survival was calculated according to life-table analyses. A step-wise multiple regression analysis was used to measure the prognostic value of the various factors. In the univariate analysis, general symptoms (p = 0.0006) psycho-social well-being (p = 0.0002) and stage of disease (p = 0.007) were the best predictive factors. In the multiple regression analyses the subjective variables, general symptoms (p less than 0.01) and psychosocial well-being (p less than 0.05) were shown to have the best predictive value for the patients' survival. PMID- 2549581 TI - [Role of low molecular weight heparin in antithrombotic treatment]. PMID- 2549584 TI - [Ki-1 non-Hodgkin's lymphoma. A multihospital study of 21 cases]. AB - The Ki-1 monoclonal antibody recognizes a specific membrane antigen of activated lymphoid cells and stains large-cell non Hodgkin's lymphomas and Hodgkin's disease. Thus, it is widely used in the diagnosis of anaplastic lymphomas. Morphologically, the Ki-1 monoclonal non Hodgkin's lymphoma are diffuse of multifocal either classical or cell anaplastic type. The clinical behaviour is similar to the rest of the high grade lymphomas, disseminated at diagnosis but may reach remission after aggressive chemotherapy. The immunophenotype showed T, B or null nature of the latter. The clinical and pathological results of our study carried out in a group of 21 cases ki-1 positive lymphomas is herewith reported. PMID- 2549583 TI - Localisation of a particulate luliberin hydrolysing activity in microsomal membranes of guinea pig brain. AB - A particulate luliberin hydrolysing enzyme has been described for guinea pig brain. Examination of subcellular fractions generated under different conditions indicated that particulate luliberin hydrolysing activity was most closely associated with the microsomal marker, rotenone-insensitive NADH cytochrome C reductase. The results obtained indicate that luliberin hydrolysing activity is not associated with synaptosomal membrane preparations and that such luliberin hydrolysing activity as is observed in synaptosomal membranes is probably the result of contamination by microsomes. The enzyme could be released from microsomes by Triton X-100 treatment and the solubilised enzyme was found to be inhibited by puromycin and sulphydryl reagents but to be unaffected by phosphoramidon, captopril, phenylmethyl sulphonyl fluoride and by chelating agents except 1,10-phenanthroline. PMID- 2549585 TI - [Mucinous carcinoma associated antigen in the blood in women with breast diseases. Preliminary results]. PMID- 2549586 TI - [Free radicals of oxygen and antioxidants in medicine]. PMID- 2549587 TI - [A 64-year-old female patient with icterus and posterior development of a picture of right ocular disease]. PMID- 2549588 TI - [Infection caused by rotavirus in children hospitalized because of gastroenteritis in Yucatan, Mexico]. AB - A one year study was performed in patients hospitalized by gastroenteritis. The purpose of this study was to determine the rate of gastroenteritis by rotavirus, as well as which subgroups and electropherotypes are the prevailing ones. Electrophoresis technique in polyacrylamide gel was used to detect ribonucleic acid of rotavirus directly obtained from feces. Results indicated that rotavirus was the cause of gastroenteritis in 28.8% of the patients studied; prevailing subgroup II. Four electropherotypes of 34 virus isolated were detected, unlike other countries where have been isolated 19 virus. PMID- 2549589 TI - [Granulomatous hepatitis caused by cytomegalovirus in an immunocompetent adult]. PMID- 2549590 TI - [Zonal variability and seasonal changes of the content of glycogen and glucose in the Mytilus mantle]. AB - Glycogen and free-glucose content in the ventral, central and dorsal parts, as well as glucose-6-phosphate phosphatase activity in mantle of Mytilus galloprovincialis Lmk. were examined. The glycogen content of mantle did not manifest asymmetrical distribution among the three parts. In the period studied, the typical glycogen content profile variation was found, being maximum in July. The tissue free-glucose content was similar in each part, and the obtained seasonal variation profile was opposite to the glycogen content, reaching the minimum in July. For every part of mantle, free-glucose/glycogen ratio showed similar monthly profiles. In each part the 50% point was found in July. Glucose-6 phosphate phosphatase activity was not found in the mantle tissue. PMID- 2549591 TI - Effect of naloxone on acute ethylic intoxication. An EEG study. AB - EEG recording has been used to evaluate in cats the effect of naloxone on acute ethylic intoxication (AEI). Naloxone was administered both before and during the course of AEI. Results of the experiment showed that administration of naloxone before the AEI potentiates the toxic effect of alcohol. However, the administration of naloxone in a continuous way along the course of AEI significantly diminished the toxic effect of alcohol. PMID- 2549592 TI - Electroencephalographic study of naloxone effects in the recovery of an acute alcoholic intoxication. AB - Experimental assays analysing EEG changes during the recovery of an acute alcoholic intoxication were carried out in three groups of cats: 1) Recovery of acute alcoholic intoxication produced by continuous intravenous perfusion of ethanol, 0.06 g/kg/min, during 20 minutes. 2) Recovery of acute alcoholic intoxication by injecting naloxone (400 micrograms/kg), just after finishing alcohol perfusion. 3) Recovery of acute alcoholic intoxication by injecting naloxone (400 micrograms/kg), 15 min after finishing perfusion. Naloxone administered after an acute alcoholic intoxication worsens the recovery of EEG parameters; 1-2 (p less than 0.05), 1-3 (p less than 0.05). PMID- 2549593 TI - Sensitivity of field isolates of Eimeria tenella to anticoccidial drugs in the chicken. AB - Thirty isolates of Eimeria tenella obtained from broiler and breeder farms were examined for their sensitivity to anticoccidial drugs. All were sensitive to robenidine, 28 were sensitive to methyl benzoquate, 25 to clopidol and 21 to nicarbazin. Most isolates were resistant or partly resistant to amprolium and dinitolmide. PMID- 2549594 TI - Pathogenicity of experimental infection with 'pneumotropic' porcine coronavirus. AB - Virus localisation and lesions were studied in 14-one-week-old piglets following combined intranasal-oral inoculation with a British isolate of 'pneumotropic' porcine coronavirus (PCV) and were compared with the effects of transmissible gastroenteritis virus (TGEV) infection in five piglets. Unlike TGEV-infected piglets, all PCV-inoculated piglets remained clinically healthy. Seroconversion was detected at seven days after inoculation. Mild bronchointerstitial pneumonia involving terminal airways was consistently present at two days after infection and thereafter. Both PCV and TGEV infected bronchiolar epithelium and alveolar macrophages but, unlike TGEV, replication by PCV in villous enterocytes was limited and did not cause villous atrophy. PMID- 2549595 TI - Oesophagogastric parakeratosis in the growing pig: effects of the physical form of barley-based diets and added fibre. AB - In four experiments a total of 288 individually fed pigs were given barley-based diets for about 100 days from about 20 kg liveweight. Fine grinding of barley increased the number and severity of oesophagogastric lesions. Pelleting a diet based on coarsely ground barley had a similar effect. Coarser grinding of the barley and substituting small proportions of oat husk, but not of bran, gave lower incidences and severities of lesions. The performance responses of the pigs differed and give a framework for deciding on the balance to be struck between optimal performance and the risk of lesion development. PMID- 2549597 TI - Preliminary characterization of cis interactions between mutations of poliovirus genome. AB - The effects of three distinct point mutations in the P3 genomic region of poliovirus type 1 (Mahoney strain) have previously been analysed. A U to C change at position 5658, which modifies the proteinase 3C, is responsible for a small plaque phenotype and defective RNA replication. An A to G change at position 7256, which modifies the replicase 3D, is responsible for temperature sensitivity. A C to U change at position 7348, which is silent at the protein level, has no apparent phenotypic effect. These three mutations were introduced into a single poliovirus genome by recombinant cDNA techniques. The resulting plurimutant virus, vHA507, was temperature-sensitive and exhibited a small plaque phenotype. Both these characters were accentuated compared to those of the viruses in which the mutations were on separate genomes. Spontaneous ts+ revertants of vHA507 arose by true reversion of the replicase mutation. The small plaque phenotype and defective RNA replication were partially suppressed in the case of these revertants, despite the fact that the proteinase mutation was still present on their genomes. This suppression was assumed to be due to the presence of the silent mutation at position 7348. This system provides a simple model of cis interactions within the poliovirus genome. PMID- 2549596 TI - BK virus (BKV) detection in urine specimens of immunocompromised patients: comparison between the DNA-DNA hybridization assay, the immunofluorescence test and the dot enzyme immunoassay. AB - Three methods used for the detection of BK virus in urine specimens, the indirect immunofluorescence test, the dot enzyme immunoassay and the DNA-DNA hybridization assay, were compared by testing specimens from 49 immunocompromised patients. All three assays were effective in detecting BK virus. The technical advantage of each of them was discussed. The immunofluorescence test was found to be the simplest one to perform; the DNA-DNA hybridization assay displayed exquisite sensitivity; and the easy reading of the dot enzyme immunoassay did not require the specialized training inherent to immunofluorescence assays. The dot enzyme immunoassay might therefore be the most practical method for screening urine specimens of immunocompromised patients, especially when the sediment is poor in cells. Conversely, the indirect immunofluorescence test might be the method of choice for checking patients with haemorragic cystitis whose urine samples usually contain large amounts of cells. PMID- 2549598 TI - Antigenic and molecular characterization of bovine rotaviruses isolated in Japan. AB - Japanese bovine rotavirus isolates, which fall into two different serotypes, were shown to belong to subgroup I and to have long RNA electrophoretypes. This study confirmed the distinction of two serotypes on the basis of a greater than 20-fold difference in neutralization titres between the homologous and heterologous reactions; however, significant one-way cross-neutralization was observed between one of the strains with bovine serotype 2 and antisera to strains with bovine serotype 1 (serotype 6 according to the unified serotyping system). When 32P labelled transcription probes prepared from Japanese strains as well as a prototype NCDV strain were hybridized with genomic double-stranded RNA from these bovine strains and from prototype human strains, a high level of homology was observed among bovine rotavirus strains; but this level of homology was not found between bovine strains and any of the prototype human strains. These results suggest that bovine rotavirus strains belong to a single genogroup that is distinct from any of the human genogroups previously identified by RNA-RNA hybridization. PMID- 2549599 TI - Detection of human cytomegalovirus DNA in liver biopsies from patients with cytomegalovirus-related liver disease. AB - Infection with human cytomegalovirus (HCMV) has been associated with severe diseases in immunologically impaired patients. Cytomegalovirus hepatitis has been frequently described in this population, but this diagnosis is still difficult. Molecular hybridization with the V EcoRI restriction fragment of human cytomegalovirus strain AD 169 has been tested upon DNA extracted from liver samples to assess the usefulness of this technique for cytomegalovirus hepatitis diagnosis. This probe was shown by the Southern technique not to hybridize with DNA extracted from cells infected with other herpesviruses or with DNA of non infected normal liver. The sensitivity was estimated to be 2 x 10(5) genomes. Twenty-five renal transplant recipients under immunosuppressive therapy and three patients having the acquired immunodeficiency syndrome were studied. In 9 out of 10 renal transplant recipients with normal liver, previous exposure to cytomegalovirus, as defined by serological tests, was not sufficient to allow positive detection by the probe. Out of 11 patients with abnormal liver, cytomegalovirus DNA sequences were shown in 5. In 2 patients with histological evidence of cytomegalovirus hepatitis, a very strong signal showed the presence of viral genomes. These results show that the Southern technique with the V EcoRI probe can be useful for the diagnosis of HCMV hepatitis and might be proposed for the detection of this viral genome in human tissues. PMID- 2549600 TI - Sources of calcium for the contraction induced by various agonists in trachealis muscle from normal and sensitized guinea pigs. AB - Active sensitization of guinea pigs resulted in an increase in the responsiveness and sensitivity of tracheal strips to CaCl2 in K+-depolarized tissue (Emax 0.81 +/- 0.22 g/mm2 and pD2 2.35 +/- 0.10 in normal vs. 0.98 +/- 0.01 g/mm2 and 3.10 +/- 0.08 in sensitized tissue; p less than 0.05), KCl (Emax 0.62 +/- 0.08 g/mm2 and pD2 1.71 +/- 0.03 in normal vs. 0.91 +/- 0.04 g/mm2 and 2.00 +/- 0.01 in sensitized tissue; p less than 0.05) and histamine (Emax 0.70 +/- 0.06 g/mm2 and pD2 5.08 +/- 0.06 in normal vs. 0.94 +/- 0.09 g/mm2 and 5.80 +/- 0.16 in sensitized tissue; p less than 0.05) but not to caffeine 10 mM (20 degrees C, indomethacin 2.8 microM). Generation of responses to these agonists in nonsensitized tissues bathed in a Ca2+-free medium resulted in the abolition of KCl-induced contraction and partial inhibition of the responses to histamine (60% inhibition) and caffeine (40% inhibition). The contraction of sensitized tracheal strips in response to histamine in 0 calcium was greater than that obtained in nonsensitized tissues (0.48 +/- 0.02 g/mm2 vs. 0.28 +/- 0.04 g/mm2, respectively; p less than 0.05). The caffeine-induced contraction of sensitized tracheae was independent of extracellular calcium. These results suggest that a greater calcium entry and/or intracellular calcium release may be an alteration underlying hyperreactivity of sensitized tissues to spasmogens. PMID- 2549601 TI - Is the desert lung syndrome (nonoccupational dust pneumoconiosis) a variant of pulmonary alveolar microlithiasis? Report of 4 cases with review of the literature. AB - Four cases of pulmonary alveolar microlithiasis are presented. These patients had heavy exposure to sand particles for long periods. This is to hypothesize that those sand particles are responsible for triggering a hyperimmune response resulting in the formation of microlithiasis. Previously expressed theories had linked microlithiasis to external irritants and the condition has similarities with desert lung syndrome, a disease definitely caused by the deposition of sand silica in the lungs. PMID- 2549602 TI - New absorbable synthetic explants for the treatment of retinal detachment. PMID- 2549603 TI - [A serological survey and histopathological comparison of HBV-associated liver diseases in southern part of Korea and west Japan]. AB - A high mortality rate due to hepatocellular carcinoma (HCC) associated with a high prevalence of hepatitis B virus (HBV) infection is a big problem in all Asian countries, including Korea and Japan. In this study, a serological survey and a histopathological comparison of HBV-associated liver diseases in Korea and in the western part of Japan were performed. In- and outpatients of Kosin University Hospital (KUH) and of UOEH Hospital were analysed serologically, and the HBsAg-positive rate among the staff of both universities were also compared. Two-hundred consecutive biopsied livers in KUH, 400 autopsied livers in Kitakyushu, and 576 livers of medicolegal autopsies in Osaka were examined histopathologically. Serological positive rates of HBsAg were 14.5% in KUH patients and 7.0% in UOEH ones respectively (P less than 0.001). Overall positive rates of HBsAg in liver tissues were 10.5% in KUH, 4.3% in Kitakyushu and 0.5% in Osaka. Primary liver cancers, most of which were HCCs, accounted for 3.4% of all inpatients in KUH and 1.9% in UOEH (P less than 0.01). HBsAg-positive rates among liver cirrhosis and HCC patients were higher in Korea than in Japan. Alcohol was thought to be a predominant cause of 36 liver cirrhosis cases out of 576 cases from Osaka, and a low positivity of HBsAg and a low association of HCC in these Osaka cases were noted. HCC patients of KUH were eleven years younger than those of UOEH on the average, and Edmondson's grade of the former was higher than the latter.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549604 TI - Hyperechoic pattern in breast cancer--its bio-acoustical genesis and tissue characterization. AB - Clinical breast echography is increasingly being used as it offers a high degree of diagnostic accuracy rate. Internal echoes of usual breast cancer is hypoechoic in the majority of cases, however hyperechoic mass lesion may appear in rare instances. This echo-pattern was seen in a 38-year-old housewife with invasive ductal carcinoma. Also, its bio-acoustical mechanism, that is, why hyperechoic pattern appears, was investigated from the viewpoint of ultrasonic tissue characterization and its related papers were reviewed. Tumor heterogeneity in cellularity such as cribriform pattern, tubular structure, solid nests and a scirrhous pattern of cancer cells may play an important role in producing the hyperechoic breast pattern. PMID- 2549605 TI - Diagnosis and management of superficial bladder cancer. AB - Superficial transitional cell carcinoma is defined as a transitional cell urothelial tumor that is confined to the mucosa, stages Ta or CIS, or with invasion of the lamina propria, T1. The initial treatment is transurethral resection with an attempt to remove all tumor. This should provide an accurate histologic grade and stage, and from this information a prognosis can be determined. The important predictive factors that correlate with a new occurrence or true recurrence and the development of a subsequent tumor with muscle invasion are a high tumor grade, lamina propria invasion, a positive cytology following resection, multifocal tumors, dysplasia or carcinoma in situ from mucosal biopsies of normal appearing urothelium, and a prior history of bladder cancer. Based on these factors, the recurrence rate varies from 30 to 80% and progression with a muscle invasive tumor up to 30%. Intravesical chemotherapy or "immunotherapy" following tumor resection has been shown to diminish the likelihood of a recurrence. Thiotepa has been used for the longest period of time. It is relatively inexpensive, safe if myelosuppression is closely monitored, and effective. Mitomycin C was more effective than Thiotepa in randomized trials, but is significantly more expensive. This has retarded its use as a first-line agent. It has been shown to eradicate persistent tumor in 30 to 40% of patients who have failed Thiotepa. Mitomycin C is also highly effective when used for prophylaxis. Intravesical bacillus Calmette-Guerin (BCG) has recently been demonstrated to be an effective intravesical therapeutic agent. It is effective both for treatment and prophylaxis. BCG is relatively safe and inexpensive.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549606 TI - Metabolism of gyrase inhibitors. AB - Since metabolites of gyrase inhibitors (including the quinolones) may contribute to or even determine the occurrence of adverse effects or interactions with other drugs, an understanding of even minor metabolic pathways is important. The extent of metabolism can be estimated by determination of the renal and nonrenal clearance of drug and by measurement of excretion of drug labeled with 14C. The principal metabolic pathways of gyrase inhibitors are piperazine ring-based reactions (formation of oxo-compounds, N-oxides, demethylation products where applicable, or ring cleavage with or without subsequent metabolic conversions) and acyl-glucuronidation at the carboxy group of the nucleus. The measurement of metabolites in plasma represents the formation, distribution, and elimination of metabolites in the body. Sensitive techniques have been developed for measuring the penetration of metabolites into fluid and tissue, and studies have been made of the metabolism of gyrase inhibitors in patients with renal failure or hepatic dysfunction. Factors that affect metabolism of gyrase inhibitors include smoking, gender, genetically determined metabolic competency, and dosing schedule. PMID- 2549607 TI - New quinolones: in vitro effects as a potential source of clinical toxicity. AB - 4-Quinolones affect mammalian cellular functions in vitro in several ways. High concentrations inhibit DNA replication, but individual genes are perhaps sensitive to lower concentrations of drug. Inhibition of cell proliferation differs widely among 4-quinolones. Ciprofloxacin and norfloxacin are the most antiproliferative, inhibiting cell growth by approximately 30% at 20 mg/L. Genotoxicity tests with 4-quinolones are probably "false-positive" as a result of increased [3H]thymidine uptake that is not related to DNA damage. Ciprofloxacin at greater than or equal to 10 mg/L causes significant strand breaks in DNA, which seemingly are quickly repaired and do not cause mutations or cancer. Production of immunoglobulin is inhibited by ciprofloxacin at a concentration of 5 mg/L, but production of the growth factor interleukin 2 (IL-2) is increased by 4-quinolones at the same concentration and is hyperinduced at higher concentrations. Thus the effects are very contradictory. Increased production of IL-2 may contribute to central nervous system adverse effects. 4-Quinolones in combination with theophylline or antiinflammatory drugs may inhibit gamma aminobutyric acid receptor binding and thereby have adverse effects on the central nervous system. Some 4-quinolones induce crystalluria, which may be nephropathic. PMID- 2549608 TI - Mode of action of the quinolone antimicrobial agents: review of recent information. AB - Recent studies concerning the mechanism of action of quinolones against DNA gyrase are reviewed. DNA gyrase is an essential bacterial enzyme known to be a primary target of quinolone agents. Quinolone-resistant alleles of both the gyrA and gyrB genes of DNA gyrase have been sequenced, and domains that affect the action of quinolones have been identified within the amino terminus of the gyrase A peptide and the midportion of the gyrase B peptide. In addition, an ATP-induced structural transition of DNA complexed with DNA gyrase was shown to be blocked by norfloxacin, but the means by which quinolones effect this change and the molecular site of quinolone binding remain unclear. Studies of structure-activity relationships of the quinolone molecule have been expanded and have included effects of quinolones on DNA gyrase. Stereochemical effects at positions 1 and 7 have been found. Substitutions at position 7 that improve potency against gram positive bacteria have also been identified. Novel mono- and three-ring structures and an isothiazolo substitution at position 3 have broadened the range of structures known to have activity. Studies of bacterial killing by quinolones have revealed additional correlations with markers of DNA damage and additional alterations in bacteria and growth conditions that affect bacterial killing. The exact events responsible for quinolone-mediated lethality, however, remain undefined. PMID- 2549610 TI - Bacterial resistance to quinolones: mechanisms and clinical importance. AB - An overview of bacterial resistance to new quinolones is presented with a consideration of mechanisms, clinical importance, and approaches to suppression of the emergence of resistance. Single-step mutation to high-level resistance occurs at a frequency of less than or equal to 10(-10) for many bacterial species but can be readily selected by serial exposure of cells to increasing drug concentrations. Two mechanisms of resistance have been identified: alteration in the target enzyme DNA gyrase and decreased drug permeation. Emergence of resistance to clinically useful quinolones thus far has been uncommon. Superinfection has been documented but also has been uncommon. Emergence of resistance appears to occur more often with certain bacterial species, including Pseudomonas aeruginosa and Staphylococcus aureus. In the special setting of cystic fibrosis, patients with P. aeruginosa infections often respond favorably despite emergence of resistance. Methods to minimize the emergence of quinolone resistance should be evaluated in an effort to preserve the clinical utility of these drugs. PMID- 2549609 TI - Quinolone uptake by bacteria and bacterial killing. AB - A review of the mechanisms of the action of 4-quinolones is presented, concentrating on the process of uptake and bacterial killing. 4-Quinolones appear to cross the gram-negative outer membrane via diffusion through outer-membrane proteins, although disruption of the normal outer-membrane barrier by 4 quinolones may facilitate diffusion via a non-porin-mediated route. Accumulation in the cytoplasm may involve influx and efflux mechanisms. The inhibition of DNA synthesis by 4-quinolones has been investigated in Escherichia coli AB1157 and found to correlate with antibacterial activity (as measured by the minimal inhibitory concentration). 4-Quinolones have other pleotropic effects on bacterial cells, such as induction of the SOS (DNA repair) response, filamentation, and direct effects on membranes. PMID- 2549611 TI - Isoenzyme analysis of Entamoeba histolytica for evaluation of pathogenicity: detection of zymodeme XIX in South America. AB - Authors report the results of the isoenzyme analysis of strains of Entamoeba histolytica isolated from international travellers. This recently developed method allows the detection of pathogenic strains by evaluating the electrophoretic mobility of cytoplasmic enzymes and was proved to be more reliable and quickly feasible than previous ones. The experience reported refers to three strains isolated from travellers coming from Ecuador, Brazil and Indonesia, respectively; the zymodeme XIX (in accordance with the Sargeaunt's classification) was identified in all the three cases. This zymodeme has been first detected in 1981 and should currently be considered rare; moreover, it has never been previously reported from the Americas. PMID- 2549612 TI - [The effect of x-ray contrast media on magnetic resonance tomography of the spinal canal]. AB - The effects of x-ray contrast media on signal intensities were studied. Relaxation times of myelographic contrast media were measured. Whereas relaxation times of Amipaque were longer, compared to CSF, the relaxation times of Duroliopaque, Omnipaque und Solutrast were significantly shorter than that of CSF. Therefore, the qualitative and quantitative effects of different contrast media on signal intensities varied. Most notable was the increase of signal intensity on T1-weighted images due to oily contrast media. PMID- 2549613 TI - [Nasopharyngeal angiofibroma]. AB - Nasopharyngeal angiofibroma is a rare benign neoplasm; it occurs almost exclusively in adolescent males. When confined to the nasopharynx, surgery is often curative. The authors report on an angiofibroma which presented with a swelling of a cheek. The tumor was located in the nasopharynx and extended to pterygopalatinal and infratemporal fossae. Radical removal of the tumor was performed without subsequent radiation. After 32 months the patient was readmitted for suspected recurrence. The removed tumor-like tissue proved to be of inflammatory origin. Two and a half years following treatment is the patient in good condition. Therapy of intracranially extending tumors as to surgery and/or radiation is discussed. PMID- 2549614 TI - Amosite mesothelioma in a cohort of asbestos workers. AB - A cohort of 820 asbestos workers with a short duration of exposure to amosite between 1941 and 1945 was followed. These men were alive five years after starting work and were observed until 1988. Seventeen cases of malignant mesothelioma (eight pleural, nine peritoneal) were found. The mean age at the onset of exposure was 33 years for men with pleural mesothelioma and 30 years for those with peritoneal mesothelioma. Chest pain was the main symptom in pleural mesothelioma and abdominal pain in peritoneal mesothelioma. Open lung biopsy was the most useful diagnostic approach for pleural mesothelioma, whereas for peritoneal mesothelioma it was exploratory laparotomy. Pleural patients died of pulmonary insufficiency, and peritoneal patients of wasting and inanition. In both groups the death certificate diagnosis was less accurate than the clinical diagnosis at death. The mean survival was 12.5 months from first symptom to death for the pleural group and 5.4 months for the peritoneal group. PMID- 2549615 TI - Exposure to silica dust in the Danish stone industry. AB - Exposure to silica dust among Danish stone workers was assessed from data collected in 1948-1980. After 1970, the exposure level was given in milligrams per cubic-meter and an exposure index (concentration of respirable dust divided by the threshold limit value for quartz) was calculated. The median index was 2.1 for the road and building material industry and 0.6 for the stonecutting industry. Crushing showed a median index of 2.6, compared to 1.0 for drilling, 0.9 for sieving, and 0.5 for cutting. The median index for the road and building material industry was higher for Bornholm than for other parts of Denmark. The median index of the stonecutting industry was 1.0 for Copenhagen and 0.5 for other parts of Denmark, and no data were available for Bornholm. Before 1970, the exposure level was given in particles per cubic meter, and few data were available on quartz content. Crushing showed the highest exposure before 1970. PMID- 2549617 TI - [Neuroendocrine regulation of macrophages]. PMID- 2549616 TI - [BRL 43694A--a 5-hydroxytryptamine receptor blocker as an antiemetic in cytostatic therapy]. AB - An open study of the new antiemetic BRL 43694A, a 5-hydroxytryptamine-3-receptor antagonist, was performed in 29 patients undergoing highly emetogenic cancer chemotherapy (25 patients received cisplatin in a dose greater than or equal to 50 mg/m2). Patients received BRL 43694A as a 30-minute infusion one hour after the administration of chemotherapy. 7 patients were treated with 40 micrograms/kg and 13 patients with 100 micrograms/kg BRL 43694A; the last 9 patients received an initial dose of 40 micrograms/kg with a provision for two additional interventional doses over 24 hours in the event of prolonged nausea or vomiting. 14 patients experienced no vomiting (48%) and 13 patients (45%) had 1-5 vomiting episodes over 24 hours following administration of the cytostatic agents. BRL 43694A did not cause major side effects. Based on our preliminary experience the new 5-HT3-receptor antagonist BRL 43694A is a potent antiemetic drug for the treatment of chemotherapy-induced nausea and vomiting. PMID- 2549618 TI - [Effects of platelet activating factor on blood platelets]. PMID- 2549619 TI - [Effects of neurohypophysis hormones on the secretion of some adenohypophysis hormones]. PMID- 2549620 TI - [Calcium: the entering pathways and mechanisms of vasoconstriction]. PMID- 2549621 TI - [Effect of atriopeptin on renal function]. PMID- 2549622 TI - [N-methyl-D-aspartate receptor channel complex]. PMID- 2549623 TI - [Putative endogenous ligands for the benzodiazepine receptor]. PMID- 2549624 TI - [1,25-Dihydroxyvitamin D3 is a new immunomodulating hormone]. PMID- 2549625 TI - [A new histaminergic receptor: H3-receptor]. PMID- 2549626 TI - [Pathogenesis and clinical aspects of primary liver cell carcinomas]. AB - The hepatocellular carcinoma is the most frequent primary liver cell carcinoma. On account of its geographical distribution with preferential occurrence in China, SE Asia and South Africa, exogenic causes are thought to be mainly responsible for its aetiopathogenesis. Besides mycotoxins, the hepatitis B virus infection (HBV) is particularly important. Integration of HBV-DNS into the hepatic cell DNS is considered to be an initiatory step in hepatocarcinogenesis. Clinically the tumour usually becomes manifest by upper abdominal complaints. Curative treatment can be promising only with very small tumours which can be detected most safely via ultrasound examination and alpha-1-fetoprotein determination. PMID- 2549627 TI - [Sonographic diagnosis of hepatocellular carcinoma. Results of a retrospective study 1983-1989]. AB - In a retrospective study of our hospital of the years 1983 to 1989 we diagnosed a hepatocellular carcinoma (HCC) in 30 patients. The diagnosis was confirmed by histology or cytology, ultrasound and clinical symptoms. The tumours presented sonomorphologically different patterns. Some were hypoechoic (n = 9), some hyperechoic (n = 16) and 5 had a mixed pattern. Nine of the hyperchoic lesions looked like a metastasis with a halo. 13 patients had multiple tumours or diffuse infiltration of the liver. Due to the variable sonomorphology only 16 patients were assumed to have a HCC, no definite diagnosis by ultrasound being possible. The concomitant cirrhosis of the liver was sonomorphically detected in 19 patients and was suspected in 2 further patients. Four patients had no cirrhosis; in 3 patients, the available information was insufficient. Because of the variable sonomorphology of HCC no definite diagnosis by ultrasound can be achieved. The combination of liver tumour and cirrhosis or elevated serumfetoprotein leads to the diagnosis. The diagnosis of HCC must be confirmed by histology. PMID- 2549628 TI - [Highly differentiated primary liver cell carcinoma. 2 case reports]. AB - The hepatocellular carcinoma (HCC) is the most frequently occurring primary hepatic tumour. Particular difficulties in diagnosis are encountered with the highly differentiated carcinoma. The imaging processes are of varying rank. Sonography and CT--and with some restrictions also scintigraphy--can represent focal findings in the liver with the highest degree of safety. With the highly differentiated tumour, angiography is less helpful, since it does not supply safe criteria of malignancy. Alpha-fetoprotein (AFP) is not a reliable parameter in case of a small highly differentiated carcinoma. Nevertheless, cirrhosis patients with the greatest risk factor (HBV, haemochromatosis and long-standing alcohol conditioned cirrhosis of many years) should be followed up and controlled by sonography and AFP determination every 6 months. PMID- 2549629 TI - [Surgical therapy of liver and bile duct tumors]. AB - The most effective surgical therapy of primary liver cancer (HCC) or proximal bile duct cancer (BDC) is radical resection, but only 20% of the patients will undergo this procedure, because the remaining patients in the advanced tumour stage or cirrhosis can be given palliative treatment only (chemo-embolisation for HCC, endoscopic or percutaneous draining with or without iridium-after-loading for BDC) or a liver transplantation (LTX), though under immunosuppression an early recurrence of the tumour is frequent. One-year survival after resection because of HCC without cirrhosis is represented by a figure of 80%, whereas with cirrhosis it is 18%; 3 years after LTX, 26% of patients are alive. Three-year survival in untreated BDC is 24%, after resection of the hilum 42%, after LTX 40%. PMID- 2549630 TI - [Sonographic-controlled puncture and its effect on further diagnosis and therapy]. AB - Results of 127 sonographically guided punctures performed during 1982 to 1987 were evaluated retrospectively and the results of coarse needle and fine needle punctures were compared. In 74 punctured focal liver changes the superiority of coarse needle biopsy was particularly evident whenever a definitive histological confirmation of the diagnosis is aimed at. No therapy was initiated in 82 of the 127 patients after the biopsy finding had been established, and surgery was performed in 22 cases. 18 patients were given cytostatic treatment, and tumour embolisation was performed twice. 3 patients with non-malignant disease were successfully treated conservatively in pancreatic abscess, intraabdominal tuberculosis and actinomycosis. PMID- 2549631 TI - Inhibition of DNA binding proteins by oligonucleotide-directed triple helix formation. AB - Oligonucleotides that bind to duplex DNA in a sequence-specific manner by triple helix formation offer an approach to the experimental manipulation of sequence specific protein binding. Micromolar concentrations of pyrimidine oligodeoxyribonucleotides are shown to block recognition of double helical DNA by prokaryotic modifying enzymes and a eukaryotic transcription factor at a homopurine target site. Inhibition is sequence-specific. Oligonucleotides containing 5-methylcytosine provide substantially more efficient inhibition than oligonucleotides containing cytosine. The results have implications for gene specific repression by oligonucleotides or their analogs. PMID- 2549632 TI - Identification by ENDOR of Trp191 as the free-radical site in cytochrome c peroxidase compound ES. AB - The chemical identity of the amino acid free-radical site that represents one of the two oxidizing equivalents stored in the H2O2-oxidized intermediate (compound ES) of the mitochondrial heme enzyme, cytochrome c peroxidase (CcP) has been sought for almost a quarter of a century. Site-directed mutagenesis alone cannot yield this answer. Low-temperature 35-gigahertz (Q-band) electron nuclear double resonance (ENDOR) spectroscopy was used to examine compound ES prepared from proteins containing specifically deuterated methionine or tryptophan, as well as the amino acid replacement Trp51----Phe. The results definitely identify the site of the radical in compound ES as tryptophan, most likely Trp191. PMID- 2549633 TI - The MHC-binding and gp120-binding functions of CD4 are separable. AB - CD4 is a cell surface glycoprotein that is thought to interact with nonpolymorphic determinants of class II major histocompatibility (MHC) molecules. CD4 is also the receptor for the human immunodeficiency virus (HIV), binding with high affinity to the HIV-1 envelope glycoprotein, gp120. Homolog-scanning mutagenesis was used to identify CD4 regions that are important in class II MHC binding and to determine whether the gp120 and class II MHC binding sites of CD4 are related. Class II MHC binding was abolished by mutations in each of the first three immunoglobulin-like domains of CD4. The gp120 binding could be abolished without affecting class II MHC binding and vice versa, although at least one mutation examined reduced both functions significantly. These findings indicate that, while there may be overlap between the gp120 and class II MHC binding sites of CD4, these sites are distinct and can be separated. Thus it should be possible to design CD4 analogs that can block HIV infectivity but intrinsically lack the ability to affect the normal immune response by binding to class II MHC molecules. PMID- 2549635 TI - Cell cycle: progression from interphase to telophase. PMID- 2549634 TI - Limbic seizures increase neuronal production of messenger RNA for nerve growth factor. AB - Nerve growth factor (NGF) produced by telencephalic neurons provides critical trophic support for cholinergic neurons of the basal forebrain. In situ hybridization and nuclease protection analyses demonstrate that limbic seizures dramatically increase the amount of messenger RNA for NGF in the neurons of the hippocampal dentate gyrus within 1 hour of seizure onset and in broadly distributed neocortical and olfactory forebrain neurons some hours later. The increased messenger RNA species is indistinguishable from messenger RNA for transcript B of the beta subunit of NGF from mouse submandibular gland. Thus, the expression of a known growth factor is affected by unusual physiological activity, suggesting one route through which trophic interactions between neurons in adult brain can be modified. PMID- 2549636 TI - Nutritional importance of pyrroloquinoline quinone. AB - Mice fed a chemically defined diet devoid of pyrroloquinoline quinone (PQQ) grew poorly, failed to reproduce, and became osteolathyritic. Moreover, severely affected mice had friable skin, skin collagen that was readily extractable into neutral salt solutions, and decreased lysyl oxidase. The identification of functional defects in connective tissue and the growth retardation associated with PQQ deprivation suggest that PQQ plays a fundamental role as a growth factor or vitamin. PMID- 2549637 TI - Role of prostaglandins and cAMP in the secretory effects of cholera toxin. AB - The role of adenosine 3',5'-monophosphate (cAMP) and prostaglandins in the pathogenesis of experimental cholera was evaluated. Fluid accumulated in the rabbit intestinal loop model after 16 hours of incubation with cholera toxin, prostaglandin E1, or prostaglandin E2, but not with membrane-permeable derivatives of cAMP or forskolin. Dibutyryl cAMP triggered a small, transient intestinal fluid accumulation response by 4.5 hours; however, the fluid was completely absorbed by 9 hours. After exposure of intestinal loops to cholera toxin, prostaglandin E was released into the intestinal lumen in a concentration dependent manner independent of cAMP. Thus, not only cAMP, but also prostaglandins may regulate water and electrolyte secretion in cholera. PMID- 2549638 TI - Inhibition of postsynaptic PKC or CaMKII blocks induction but not expression of LTP. AB - Long-term potentiation (LTP) of synaptic transmission is a widely studied cellular example of synaptic plasticity. However, the identity, localization, and interplay among the biochemical signals underlying LTP remain unclear. Intracellular microelectrodes have been used to record synaptic potentials and deliver protein kinase inhibitors to postsynaptic CA1 pyramidal cells. Induction of LTP is blocked by intracellular delivery of H-7, a general protein kinase inhibitor, or PKC(19-31), a selective protein kinase C (PKC) inhibitor, or CaMKII(273-302), a selective inhibitor of the multifunctional Ca2+-calmodulin dependent protein kinase (CaMKII). After its establishment, LTP appears unresponsive to postsynaptic H-7, although it remains sensitive to externally applied H-7. Thus both postsynaptic PKC and CaMKII are required for the induction of LTP and a presynaptic protein kinase appears to be necessary for the expression of LTP. PMID- 2549639 TI - Recognition of thymine adenine.base pairs by guanine in a pyrimidine triple helix motif. AB - Oligonucleotide recognition offers a powerful chemical approach for the sequence specific binding of double-helical DNA. In the pyrimidine-Hoogsteen model, a binding size of greater than 15 homopurine base pairs affords greater than 30 discrete sequence-specific hydrogen bonds to duplex DNA. Because pyrimidine oligonucleotides limit triple helix formation to homopurine tracts, it is desirable to determine whether oligonucleotides can be used to bind all four base pairs of DNA. A general solution would allow targeting of oligonucleotides (or their analogs) to any given sequence in the human genome. A study of 20 base triplets reveals that the triple helix can be extended from homopurine to mixed sequences. Guanine contained within a pyrimidine oligonucleotide specifically recognizes thymine.adenine base pairs in duplex DNA. Such specificity allows binding at mixed sites in DNA from simian virus 40 and human immunodeficiency virus. PMID- 2549640 TI - Primary structure of the beta subunit of the DHP-sensitive calcium channel from skeletal muscle. AB - Complementary DNAs for the beta subunit of the dihydropyridine-sensitive calcium channel of rabbit skeletal muscle were isolated on the basis of peptide sequences derived from the purified protein. The deduced primary structure is without homology to other known protein sequences and is consistent with the beta subunit being a peripheral membrane protein associated with the cytoplasmic aspect of the sarcolemma. The protein contains sites that might be expected to be preferentially phosphorylated by protein kinase C and guanosine 3',5' monophosphate-dependent protein kinase. A messenger RNA for this protein appears to be expressed in brain. PMID- 2549641 TI - Pathogenesis of congenital infection with three diverse viruses: varicella-zoster virus, human parvovirus, and human immunodeficiency virus. PMID- 2549642 TI - Fine needle aspiration cytology of metastatic sarcoma involving the thyroid. AB - We have presented two cases of sarcoma, namely malignant fibrous histiocytoma and leiomyosarcoma, metastatic to the thyroid. Both were diagnosed clinically by fine needle aspiration cytology. PMID- 2549643 TI - Cyclic Cushing's disease in association with a pituitary stone. AB - We have documented what we believe to be the first reported case of a pituitary stone in a patient with pituitary-dependent Cushing's disease. Pituitary stones have been reported exclusively in growth-hormone-producing or prolactin-producing pituitary adenomas. Our patient's ACTH and serum cortisol levels cycled for 15 months and then resolved spontaneously. A CT scan of the head showed calcification of pituitary tissue. Pituitary stones may occur in association with Cushing's disease. We hypothesize that the spontaneous resolution of the cyclic Cushing's disease was due to destruction and ultimate calcification of abnormal pituitary tissue. PMID- 2549644 TI - Acute Epstein-Barr virus myocarditis simulating myocardial infarction with cardiogenic shock. AB - We have reported the case of a 38-year-old white woman with substernal chest pain, hypotension, and ECG changes suggesting acute anterior myocardial infarction. Cardiac catheterization revealed no coronary artery pathology, but severe global hypokinesia was noted on left ventriculogram and endomyocardial biopsy revealed myocytic degeneration and mononuclear cell infiltration consistent with acute viral myocarditis. Viral serologies confirmed a recent Epstein-Barr virus infection. PMID- 2549645 TI - Epstein-Barr virus causing encephalitis in an elderly woman. AB - Although infectious mononucleosis commonly afflicts young persons, it is now being described more frequently in the elderly. Neurologic manifestations are uncommon, and encephalitis is rare, especially in elderly patients. We have reported a case of acute Epstein-Barr virus infection causing acute encephalitis in a 64-year-old woman. Despite the severity of encephalitis, the patient recovered fully after several weeks. PMID- 2549646 TI - Studies on the Japanese encephalitis vectors in Amphoe Muang, Chiang Mai, Northern Thailand. AB - From February 1987 to January 1988, biological and ecological studies were made to obtain the basic knowledge of Japanese encephalitis vectors, Culex tritaeniorhynchus, Cx. gelidus, and Cx. fuscocephala, in Amphoe Muang, Chiang Mai, northern Thailand. The following results were found. Peaks in the population densities of the vectors as measured four times a month, by UV-light trap and human-baited trap collections, occurred during rainy season. The JE vectors in rural areas showed a sharp rise in the population in July when most of the rice fields were ploughed and a marked decline in mosquito population densities occurred after transplanting in August when the fields were flooded. The average number of larvae plus pupae per m2 in rice fields was highest in July when the fields were ploughed, but in the period from transplanting to harvesting (August to November), the densities were very low. Daily survival rates of the adult females, as estimated from parous rates, were mostly as high as 0.7 throughout the year. PMID- 2549647 TI - [Benign multilocular cystic nephroma. Description of 1 case]. AB - The case of a multiple kidney cyst in an adult patient is described. The cyst had caused parenchymal atrophy by blocking the excretory ways. The pathological anatomy of the lesion is described and the aetiopathogenic hypotheses found in the literature are reported. PMID- 2549648 TI - The endorectal ileal pullthrough procedure in patients with ulcerative colitis and familial polyposis with carcinoma. AB - Since 1977, 196 patients (177 with ulcerative colitis and 19 with familial polyposis) have undergone colectomy, mucosal proctectomy and endorectal ileal pull-through with or without an ileal reservoir (PTR) at UCLA Medical Center. Fourteen of the patients (7.1 per cent) had carcinoma of the colon or rectum at the time of operation; 12 had colitis and two, polyposis. Another 40 patients had mucosal dysplasia. Only five of the 14 patients with carcinoma were diagnosed before operation despite close surveillance by gastroenterologists. The mean duration of colitis before the diagnosis of carcinoma was made was 17 years; the mean age that the carcinoma was identified was 38 years. Eleven of the 12 patients with colitis had universal involvement. Two patients with colitis and carcinoma who underwent colectomy and PTR died a mean of 30.5 months postoperatively of metastatic disease. Twelve patients with carcinoma (ten with colitis and two with polyposis) are alive a mean of 29 months postcolectomy and PTR; two of these have received chemotherapy. The low mortality (0.4 per cent) and good clinical results after colectomy and the PTR procedure and the unexpectedly high incidence of carcinoma and mucosal dysplasia among patients referred for operation suggest that surgical treatment should be considered at an earlier stage than the current general practice, particularly in patients at high risk (mucosal dysplasia, pancolitis and duration of more than ten years). PMID- 2549649 TI - [Results of local radiotherapy and modified total-brain irradiation in astrocytoma]. AB - A retrospective evaluation was made on the results of primary or postoperative radiotherapy in 162 patients with grade I to IV astrocytomas. The greatest part of the overall group was built by 113 patients suffering from glioblastoma multiforme. For the survival analysis, the prognostic importance of the following factors was investigated: sex, age, surgery, radicality of surgery, grading according to Kernohan, tumorous lesions in central structures, and target volume of radiotherapy. The prognosis is first of all dependent on the operability, the grading, and the age of the patient. The survival of patients with malignant gliomas is not influenced by the radicality of the operation and the extension of the target volume to the whole brain (total-brain irradiation and boost). Thus in the treatment of malignant astrocytomas, the less extensive surgical and radiotherapeutical intervention seems an adequate method to manage this disease. PMID- 2549650 TI - In vivo ESR dosimetry in total body irradiation. AB - Total body irradiation (TBI) using high doses (about 10 Gy) with photons in the range between 1 and 10 MV combined with intensive chemotherapy has been used successfully in treatment of acute and chronic leukemia before bone marrow transplantation. One of the principal international guidelines in TBI is to use in vivo dosimetry in order to compare the prescribed dose with that absorbed. The use of in vivo dosimetry is also useful as a retrospective evaluation of any deviation from the prescribed dose greater than +/- 5% for relevant parts of the body, especially in the lung and in other organs at risk. In this paper, Electron Spin Resonance (ESR), using alanine dosimeters, is demonstrated to be a powerful tool for absorbed dose evaluation in TBI by detection of free radicals produced in alanine by ionizing radiation. In this study, we present the results obtained using ESR dosimetry in eleven patients undergoing TBI. The major advantages appear to be: 1. the ESR signal in alanine dosimetry is stable for years without fading: 2. the detection of the ESR signal does not destroy the information and so enables a retrospective judgment of the TBI plan adopted. PMID- 2549651 TI - Phencyclidine and sigma receptors in rat spinal cord: binding characterization and quantitative autoradiography. AB - These experiments were designed to compare phencyclidine (PCP) and sigma (sigma) receptor binding sites in the rat spinal cord by using receptor binding and autoradiographic techniques. Binding sites for 3H-TCP (3H-1-[1-(2 thienyl)cyclohexy]piperidine), a PCP receptor agonist, and (+)3H-3-PPP (3H-(+)-3 (3-hydroxyphenyl)-N-(1-propyl)piperidine), a sigma receptor agonist, in the rat spinal cord were shown to represent two populations of recognition sites. Inhibition studies revealed that ligands with high affinity for the PCP receptor (MK-801 and PCP) were potent competitors at 3H-TCP binding sites whereas the putative sigma receptor ligands (+/-)pentazocine and haloperidol were potent competitors at (+)3H-3-PPP binding sites. The autoradiographic distribution of 3H TCP and (+)3H-3-PPP binding sites in adjacent sections of rat spinal cord demonstrated the occurrence of two distinct populations of binding sites. 3H-TCP binding sites were localized primarily in laminae I and II in cervical and thoracic spinal segments. Binding sites in lamina I decreased in density along a rostral to caudal gradient in the spinal cord. The highest density of (+)3H-3-PPP binding sites was found in the ventral horn (lamina VIII and IX) and over perikarya in dorsal root ganglia. Significantly elevated densities of (+)3H-3-PPP binding sites were also found in lamina X within thoracic and lumbar segments and in the intermediolateral cell column. The results of the present study show that PCP and sigma receptor binding sites are differentially localized in the rat spinal cord and suggest that separate binding sites exist for PCP and sigma agonists. PMID- 2549652 TI - Brain abnormalities induced by murine cytomegalovirus injected into the cerebral ventricles of mouse embryos exo utero. AB - Murine cytomegalovirus (MCMV) was injected into the cerebral ventricle of mouse embryos on day 13 of gestation after exposing the embryos out of the uterus in the abdominal cavity of the mother. The embryos were allowed to develop to day 18 of gestation, then taken out from the abdominal cavity. Macroscopically, there were four expanded and three distorted brains out of 19 surviving embryos, whereas no brain abnormality was noticed in 13 embryos injected with culture medium instead of MCMV in the same way. Histopathological examination showed hydrocephalic lesions with strong dilatation of the ventricles and atrophy of the cerebral cortex, and inflammatory lesions with granulomatous proliferation of the ventricular walls with disappearance of the cortical zonation. Immunohistochemically, MCMV-induced nuclear antigen-positive cells were frequently observed in the wall of the ventricles and occasionally scattered in the cerebral cortex, white matter, and the nucleus basalis. Some fetuses injected with MCMV in the same way were recovered from the abdominal cavities on day 18 of gestation and transferred to foster nurse mothers. They showed massive cerebral necrosis after feeding for 9 days after birth. Brain abnormalities of mouse embryos after intraventricular injection with MCMV may provide an experimental model of brain damage induced by congenital cytomegalovirus infection. PMID- 2549653 TI - Diagnosis of pulmonary disease in human immunodeficiency virus infection: role of transbronchial biopsy and bronchoalveolar lavage. AB - The value of transbronchial biopsy and bronchoalveolar lavage was assessed in the diagnosis of pulmonary disease in patients infected with the human immunodeficiency virus (HIV). Seventy four transbronchial biopsy and 66 bronchoalveolar lavage specimens (60 paired specimens) from 80 examinations in 64 patients were reviewed. Pneumocystis carinii was the most common pathogen isolated (43 patients). Bronchoalveolar lavage was superior to transbronchial biopsy for the diagnosis of this pathogen, the sensitivities being 90% and 56%. Cytomegalovirus was identified three times by lavage and once by transbronchial biopsy. Neither method detected Kaposi's sarcoma in the one patient shown to have it by open lung biopsy. The complication rate in a concurrent study of bronchoscopy with transbronchial biopsy in 74 consecutive HIV positive patients was 22%. This study does not support the use of transbronchial biopsy in these patients. PMID- 2549654 TI - Comparison of two experimental thrombosis models in rats effects of four glycosaminoglycans. AB - Two experimental thrombosis models in rats have been compared with regard to the composition of the formed thrombi and the effects of various treatments on thrombus formation. In the first model thrombosis is induced in the vena cava by a combination of venous stasis and hypercoagulability; these thrombi consist merely of red cells and fibrin with only a few platelets. In the second model thrombosis is induced in an arterio-venous shunt in which the formed thrombi consist of red cells, fibrin and a large amount of platelet aggregates adhering to the foreign material. Antiplatelet serum and acetylsalicylic acid, which reduce blood platelet activity, inhibited thrombus formation only in the arteriovenous shunt model. Dicumoxane, an oral anticoagulant, was active in both models. The glycosaminoglycans heparin, Org 10172, Fragmin and the pentasaccharide, representing the AT-III binding sequence of heparin, were active in both models. However, there were qualitative and quantitative differences between the effects of the glycosaminoglycans suggesting differences in their modes of action. PMID- 2549655 TI - Clotting of whole human blood induces cysteinyl-leukotriene formation. AB - Using radioimmunoassay techniques we studied the formation of the 5-lipoxygenase derived cysteinyl-leukotrienes (LT) in comparison to the cyclooxygenase product thromboxane (TX) B2 in whole human blood allowed to clot at 37 degrees C in vitro. Spontaneous clotting resulted in a time-dependent release of smaller amounts of cysteinyl-LT as well as release of large amounts of TXB2 into the serum. Cysteinyl-LT were characterized by their immunoreactive behaviour and their biological activity in the guinea pig ileum bioassay, an effect which could be antagonized by the SRS-A antagonist FPL 55712 (0.38 microM). By reversed phase HPLC cysteinyl-LT in the serum were identified as a mixture of LTC4, LTD4 and LTE4. At 90 and 120 min part of the immunoreactive material consisted of the omega-oxidized metabolite 20-OH-LTE4. Almost complete inhibition of cyclooxygenase activity by indomethacin (2.8 microM) did not affect cysteinyl-LT formation by clotting whole human blood in vitro nor did activation of platelets by compounds such as the TX mimetic U 46619 (10 microM), platelet-activating factor (PAF, 1 microM) or thrombin (3 IU/ml). In contrast, the lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA, 10 microM), the Ca2+-chelating anticoagulants trisodium citrate (10 microM) and edetate disodium (EDTA, 5.4 mM) as well as the functionally unrelated heparin (20 IU/ml) significantly inhibited the formation of cysteinyl-LT as well as of TXB2. Thus, an event related to the process of clotting of whole human blood appears to be able to induce cysteinyl LT formation in amounts which might be functionally relevant during thromboembolic events. PMID- 2549656 TI - [Forced alkalic diuresis. Still a current therapy in poisoning]. PMID- 2549657 TI - Histological changes in the skin of Rana pipiens produced by metabolic alkalosis. AB - The frog skin has been shown to excrete various electrolytes, the rates can be altered by varying metabolic conditions. The present study was performed to determine if metabolic alkalosis results in histological changes in the skin that are characteristic of this state. Rana pipiens were loaded with NaHCO3 and skin biopsies obtained (I). These biopsies were compared with biopsies from either control, unloaded frogs (II), or from NaCl loaded (III) frogs. In blind studies of microscopic sections, 13 of 15 biopsies of a mixture of I and II were correctly diagnosed, and similarly, 18 of 20 of I and III were correctly diagnosed (P = 0.0037, and 0.0002, respectively). The changes due to NaHCO3 treatment included; (1) an abundance of large euchromatin cells on or near the surface; (2) changes in the basal cell layer with elongation and rotation of the nuclei; (3) lighter cells in the spinosal layer; and, (4) sometimes the skin became thicker. We conclude that metabolic alkalosis results in characteristic histological changes in the skin, and that this is probably related to the ability of the skin to excrete bicarbonate. PMID- 2549658 TI - Fatal acute interstitial pneumonia induced by low-dose doxorubicin and vindesine. AB - A 53-year-old female smoker with small cell carcinoma of the lung with cerebral metastases was initially treated with whole brain radiation with favorable responses. She developed fulminant and fatal interstitial pneumonia following administration of 30 mg of doxorubicin and 6 mg of vindesine. Histopathology revealed that the interstitial pneumonia was compatible with a drug-induced pneumonia. PMID- 2549659 TI - Synthetic approach to analogues of 19-norsteroids with an acyclic side chain. AB - Racemic 14 beta-hydroxy-3-methoxy-8 alpha,9 alpha-1,3,5(10)-estratriene-17-one (I), obtained by total synthesis, was converted into a derivative with alkoxycarbonyl-ethylenic side chain, rac-(20E)-21-methoxycarbonyl-19-nor-8 alpha,9 alpha-pregna- 1,3,5(10),20-tetraene-3,14 beta-diol 3-methyl ether (XII) using two Wittig reactions. Analogous derivatives of 5 alpha-androstane were prepared as synthetic models. In the estrane series the stereochemistry of attachement of the side chain in position 17, biological activity of some compounds, and their chromatographic properties were investigated. PMID- 2549661 TI - Does muscle activation occur by direct mechanical coupling of transverse tubules to sarcoplasmic reticulum? AB - Our knowledge of the physiological and biochemical constituents of skeletal muscle excitation has increased greatly during the last few years but this has not led to a consensus of the physiological mode of muscle activation. Three hypotheses of transmission, involving either transmitter-receptor interaction or direct mechanical coupling, are still under active consideration. The hypothesis of direct mechanical coupling currently being evaluated proposes that the dihydropyridine receptor in the transverse tubules serves as a voltage sensor that communicates directly with the junctional foot protein/Ca2+ channel of sarcoplasmic reticulum to initiate opening of the channel. PMID- 2549660 TI - Differential up- and down-regulation of type I and type II receptors for adrenocorticosteroid hormones in mouse brain. AB - Adult female mice were adrenalectomized and ovariectomized and the concentration of Type I and Type II receptors in whole brain, kidney, and liver cytosol determined at various time thereafter by incubation with [3H]aldosterone (+ RU 26988 to prevent binding to Type II receptors) or [3H]dexamethasone, respectively. Type I receptor binding in brain was found to undergo a dramatic biphasic up-regulation, with levels six times that of intact levels by 24 h post surgery and a doubling again by 4-8 days post-surgery. By 16 days, however, Type I specific binding had returned to intact levels. Similar, but less dramatic fluctuations were seen in kidney and liver, whereas much smaller fluctuations were seen for Type II receptors in all three tissues. In a follow-up study with Scatchard analyses we observed a similar transient up- and down-regulation in maximal binding for Type I, and to a lesser extent Type II receptors in all three tissues. As expected, the apparent binding affinity for both receptors increased after surgical removal of competing endogenous steroids. Radioimmunoassays revealed that plasma concentrations of corticosterone were reduced to near undetectable levels by 24 h post-surgery. A direct comparison of male and female mice revealed no sex-related differences in Type I receptor binding capacity fluctuations in brain cytosol after adrenalectomy-gonadectomy. Lastly, treatment with exogenous aldosterone or corticosterone was found to prevent adrenalectomy gonadectomy-induced up-regulation of Type I and, to a lesser extent, Type II receptors in brain. Somewhat surprisingly, the potency of these two adrenocorticosteroids appeared to be very similar for both receptor types. PMID- 2549662 TI - Group I introns: do they only go home? PMID- 2549663 TI - Are intron-encoded specific endonucleases responsible for nonhomologous recombination? PMID- 2549665 TI - Bivalent ligands and the message-address concept in the design of selective opioid receptor antagonists. AB - Metabolically stable receptor antagonists that are subtype selective are indispensable pharmacological tools. In this article, Philip Portoghese describes the bivalent ligand approach to drug design which has resulted in the development of several highly selective non-peptide opioid receptor antagonists, such as the kappa-selective norbinaltorphimine and the delta-selective naltrindole. Models used resemble Schwyzer's message-address concept which originally described the recognition elements of peptide hormones; their success augurs well for the possibility of altering antagonist selectivity in a predictable fashion by simulating a portion of the address peptide component with a rigid nonpeptide moiety. PMID- 2549664 TI - Similarities in mode and sites of action of sarafotoxins and endothelins. PMID- 2549666 TI - Non-peptide ligands for peptide receptors. AB - Considerable progress has been made in synthesizing peptide analogs with improved stability for probing function of peptide-receptor systems. However, since peptide ligands are usually unsuitable for development as potent orally active long-duration therapeutic agents, considerable research effort is being directed to the development of non-peptidal ligands. Roger Freidinger compares the lead compounds that have now been described in the opioid, CCK-gastrin and angiotensin II fields, and discusses the progress that has been made in other receptor fields. Most of the successes have been achieved through screening natural sources and synthetic collections. Rational design based on the natural peptide ligand has been more difficult, although ACE inhibitors have been effectively developed from a nonapeptide lead. PMID- 2549667 TI - Human fibroblast interferon adjuvant to CO2 laser in the treatment of recurrent juvenile laryngeal papillomatosis: experience with 7 cases. AB - Preliminary results of adjuvant human fibroblasts interferon (IFN beta) given after CO2 laser excision in recurrent laryngeal papillomatosis in 7 adult patients are reported. Diagnostic procedure included histologic and immunohistochemical investigations to demonstrate the presence of viral cytopathic effect and for characterization of the virus. All patients underwent CO2 laser excision under general anesthesia followed by administration of IFN beta intramuscularly at the dose of 4 x 10(6) IU/day for 10 consecutive days. In the presence of complete remission, patients were followed without further therapy; in the presence of partial remission, a new combined treatment was established. All patients had a complete remission after combined treatment, but 4 subsequently developed recurrences. Treatments were always well tolerated; even cirrhotic patients showed no side effects. PMID- 2549668 TI - Small-cell lung cancer: importance of fiberoptic bronchoscopy in the evaluation of complete remission. AB - The authors report their eight-year experience on the methodical of fiberbronchoscopy in the evaluation of complete remission in 140 patients affected by small-cell lung cancer. The higher reliability of fiberbronchoscopy than of standard chest X-ray is emphasized. PMID- 2549669 TI - Acute bleeding from leiomyoblastoma of the greater omentum. A case report. AB - Extragastric leiomyoblastomas are extremely rare. This is the sixth reported case of leiomyoblastoma of the greater omentum, and the only one with acute major bleeding which required an emergency operation. Leiomyoblastomas are low-grade malignant tumors, and especially the gastric ones have an excellent prognosis after surgical removal. Extragastric leiomyoblastomas have a higher rate of dissemination. However, none of the previously reported omental leiomyoblastomas showed signs of metastatic spread at the time of operation. The present patient is alive and well 18 months post-operatively. PMID- 2549670 TI - [Herpesvirus and acute peripheral facial paresis]. PMID- 2549671 TI - Community help for women with breast cancer after discharge from hospital. PMID- 2549672 TI - Diagnosis of feline leukemia virus and feline immunodeficiency virus infections. AB - Feline leukemia virus is an oncogenic retrovirus that can result in a wide variety of neoplastic and non-neoplastic diseases, including immunosuppression. Diagnosis of FeLV infection can be achieved by several methods, including virus isolation; IFA assay of a peripheral blood smear; and detection of a viral protein (called p27) by ELISA testing of whole blood, plasma, serum, saliva, or tears. Commercially available ELISA kits have revolutionized FeLV testing and have become very popular as "in-house" procedures. This article discusses the interpretation of ELISA results and compares them with IFA assay findings. Feline immunodeficiency virus is a lentivirus that causes immunosuppression, but not neoplasia, in cats. It originally was called feline T-lymphotropic lentivirus. Differentiating FIV infection from the immunosuppressive type of FeLV infection requires virus isolation or serology. The most rapid method for diagnosis of FIV infection is ELISA testing for antiviral antibody. PMID- 2549673 TI - Control of Aujeszky's disease. PMID- 2549674 TI - Outbreak of kennel cough in Norway. PMID- 2549675 TI - A parvovirus-like agent associated with psittacine beak and feather disease. PMID- 2549676 TI - Infection with porcine respiratory coronavirus does not fully protect pigs against intestinal transmissible gastroenteritis virus. AB - Eight nine-week-old specific-pathogen-free pigs which had been infected with the transmissible gastroenteritis virus (TGEV)-related porcine respiratory coronavirus (PRCV) and four uninfected littermates were challenged with TGEV. The previous PRCV infection failed to protect them against the enteric TGEV infection. Virus excretion in faeces was detected by an ELISA in all the pigs for three to six consecutive days after inoculation. Although little diarrhoea was observed, the infection extended through much of the small intestine of one of the previously infected pigs four days after inoculation. Challenge with TGEV caused a secondary neutralising antibody response. By using a peroxidase conjugate of a monoclonal antibody which recognises a specific antigenic site on TGEV, antibodies against TGEV could be distinguished from antibodies against PRCV in an ELISA blocking test. PMID- 2549677 TI - Comparison of two ELISAs for detecting antibodies to glycoprotein I of Aujeszky's disease virus. PMID- 2549678 TI - Sheep herpesvirus does not increase susceptibility to pneumonic pasteurellosis. PMID- 2549679 TI - Characterization of monoclonal and polyclonal antibodies to bovine enteric coronavirus: establishment of an efficient ELISA for antigen detection in feces. AB - Monoclonal antibodies to bovine enteric coronavirus (BEC) were produced. Additionally, polyclonal antibodies were made in rabbits and guinea pigs and extracted from the yolk of immunized hens. The antibodies were characterized by neutralization test, hemagglutination inhibition test, enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Neutralizing antibody titers of polyclonal antisera ranged from 1:1280 to 1:40,000. Only one out of 908 hybridoma colonies tested secreted antibodies with neutralizing activity. By ELISA, polyclonal sera exhibited high background reactions that could be significantly reduced by treatment with kaolin in the case of rabbit sera. Attempts to establish an ELISA for BEC antigen detection based on polyclonal sera failed due to low sensitivity and specificity. Optimal results were achieved when a mixture of two monoclonal antibodies was coated onto microplates for antigen capture, while rabbit hyperimmune serum served as detecting antibodies in an indirect assay. The combination of the two monoclonal antibodies did not increase sensitivity synergistically, but in a compensatory fashion, probably because of epitope differences between BEC field strains. PMID- 2549680 TI - Comparative analysis of monoclonal antibodies against pestiviruses: report of an international workshop. AB - Thirty-three pestivirus strains were grown in cell culture and characterized by immunostaining with 19 monoclonal antibodies (MAbs) raised against hog cholera virus (HCV), with 42 MAbs against bovine viral diarrhoea virus (BVDV) and with 13 MAbs against border disease virus (BDV). Seven MAbs reacted with all pestivirus strains tested, eight MAbs detected only the seven HCV strains, three detected only the 16 BVDV strains. No MAb was found that was specific for BDV. BVDV and BDV strains were broadly cross-reactive with the MAbs, indicating a close relationship between these two species, whereas HCV strains were characterized as distinct from BVDV and BDV. PMID- 2549681 TI - Quantitation, biological and physicochemical properties of cell culture-adapted porcine epidemic diarrhea coronavirus (PEDV). AB - The porcine epidemic coronavirus (PEDV), tentatively classified as a coronavirus, was adapted to Vero cells and a plaque test developed for infectivity titration, allowing us to test the biological and biophysical properties of the virus. Growth kinetics showed peak titers of 10(5.5) plaque-forming units ml-1 15 h after infection. Filtration experiments and electron microscopy revealed a particle diameter between 100 and 200 nm. The buoyant density of the virus was 1.18. The particle lost its infectivity on treatment with lipid solvents. Virus replication could not be inhibited by 5-iodo-2'-deoxyuridine. PEDV was moderately stable at 50 degrees C, but heat sensitivity was not altered by divalent cations. At 4 degrees C, the virus was stable between pH 5.0 and 9.0, but at 37 degrees C stability was restricted to the pH range 6.5-7.5. Viral infectivity was not impaired by ultrasonication or by multiple freezing and thawing. PEDV was not neutralized by transmissible gastroenteritis virus antiserum. On the basis of the tests carried out, PEDV is a pleomorphic, enveloped RNA virus with a particle diameter of approximately 150 nm and a buoyant density of 1.18. Infectivity depends on the presence of trypsin, and infected cells show a tendency to fuse and to form syncytia. All of these properties, as well as its physicochemical characteristics, allow PEDV to be classified as a coronavirus. PMID- 2549682 TI - Iscom of viral envelope proteins protects against Aujeszky's disease. AB - An immunostimulating complex (iscom) containing the envelope proteins of pseudorabies virus (PRV) was prepared and its efficacy was evaluated in two experiments on sheep. In the first experiment, sheep were intramuscularly (i.m.) or intradermally (i.d.) vaccinated with PRV iscom doses varying between 1 and 81 micrograms. The vaccination was repeated on Day 21 and the animals were exposed to challenge infection by subcutaneous inoculation of 1000 TCID50 of the virulent Phylaxia strain on Day 35 after first vaccination. In the second experiment, sheep were i.m. vaccinated with single doses of iscom varying between 1 and 27 micrograms and challenge-infected on Day 14. It was found that: (1) the i.d. administration of PRV iscom has no advantage over i.m. administration (2); a single dose of greater than or equal to 3 micrograms of PRV iscom provided protection against the disease. In immunoblots, viral proteins of molecular masses 120, 109, 55, 53 and 32 kDa were detected with the sera obtained from iscom-vaccinated and subsequently challenge-infected sheep, but not with sera from sheep which were iscom-vaccinated only. The above findings indicated that: (1) by using iscom technology, potent subunit vaccines can be prepared to prevent Aujeszky's disease; (2) the selective incorporation of viral envelope proteins into iscoms gives the opportunity to discriminate between iscom-vaccinated and naturally infected animals. PMID- 2549683 TI - Evaluation of techniques to demonstrate foot-and-mouth disease virus in bovine tongue epithelium: comparison of the sensitivity of cattle, mice, primary cell cultures, cryopreserved cell cultures and established cell lines. AB - Tongue epithelia infected with each of the 7 serotypes of foot-and-mouth disease virus (FMDV) were used to evaluate in vivo and in vitro systems for the detection of FMDV. Cattle inoculated by the intradermal route in the tongue (IDL) and suckling mice inoculated intraperitoneally were compared for susceptibility to FMDV with freshly prepared bovine thyroid cell cultures; cultures from cryopreserved bovine thyroid, bone marrow, mammary gland, myocardium, tongue, ovary and kidney cells; cultures from cryopreserved embryonic ovine kidney, newborn ovine kidney, ovine testicle, bone marrow, and chloroid plexus cells; and the continuous porcine kidney cell lines MVPK-1 and S6. The mean titers determined for each serotype in each system were statistically compared. The FMDV titers obtained in freshly prepared bovine thyroid cell cultures and by cattle IDL inoculation were the highest and were statistically indistinguishable. The titers obtained by suckling mouse inoculation were significantly lower than the titers obtained in thyroid cultures for serotypes A, C, Asia 1, and SAT 3. The cattle IDL assay was significantly more sensitive than the mouse assay for serotype A. The cell cultures from the cryopreserved newborn ovine kidney and embryonic ovine kidney were significantly less susceptible to serotype Asia 1 when compared with the fresh bovine thyroid cultures, but not significantly different when compared with the cattle assay for all serotypes. Cryopreservation of bovine thyroid cells directly after trypsinization resulted in the loss of susceptibility to FMDV serotype SAT 2. The other cryopreserved cell culture systems exhibited no or minimal susceptibility to all 7 serotypes, or exhibited considerable inconsistency. The established cell lines MVPK-1 and S6 were not susceptible to serotype A, and were less sensitive to serotype C than other culture systems. Quality control of cell cultures used to evaluate field specimens for FMDV was critical. The cell cultures of cryopreserved ovine kidney cells provided the most practical diagnostic system. PMID- 2549684 TI - The role of the reticuloendothelial system in the immunopathology of Marek's disease. AB - The role of macrophages in immunity against Marek's disease (MD) was studied. Chickens of one group were subjected to depletion of macrophages using repeated doses of Francil amorphous silica and those of another group were subjected to activation of macrophages using repeated doses of brewer's thioglycollate broth. Chickens of a third group were vaccinated with herpesvirus of turkeys FC 126 vaccine followed by depletion of macrophages. Chickens of these three groups, as well as groups of healthy unvaccinated and healthy vaccinated chickens, were challenged with virulent MD virus. A sixth group of healthy uninfected chickens was kept as a control. The results, based on clinical signs, gross and histopathological studies and agar gel precipitation test (AGPT) for antibodies, indicated that activation of macrophages enhanced immunity against MD and depletion of macrophages had the opposite effect. The protective effect of vaccination against MD was also lowered by depletion of macrophages. The results of AGPT indicated retardation of MD virus replication by macrophage activation and the reverse on depletion. PMID- 2549685 TI - Antibodies to foot-and-mouth disease virus infection associated (VIA) antigen: use of a bioengineered VIA protein as antigen in an ELISA. AB - An enzyme-linked immunosorbent assay (ELISA) to detect antibodies to foot-and mouth disease (FMD) virus infection associated (VIA) antigen (viral RNA polymerase) in cattle sera, was developed using a bioengineered VIA (BioVIA) protein antigen. Compared with the classical immunodiffusion test, with viral RNA polymerase purified from infected cell cultures as antigen, this ELISA was more sensitive. However, depending on the cattle population examined, sera with antibodies to viral RNA polymerase, probably due to infection with other picornaviruses, were detected. Despite these observations, the ELISA using BioVIA provided a rapid answer as to whether or not FMD virus circulated in a given herd of cattle. The main advantage of this ELISA is its absolute safety, since in no step of the antigen production was infectious or uninfectious FMD virus involved. The test can therefore be performed under normal laboratory conditions and no isolation units are needed as they are for the immunodiffusion test. PMID- 2549686 TI - Serological detection of multiple retroviral infections in cattle: bovine leukemia virus, bovine syncytial virus and bovine visna virus. AB - Individual experimental animals used in our studies on bovine leukemia virus (BLV) are routinely screened for the presence of antibodies to the three bovine lymphotropic retroviruses. We utilized these screening methods to examine frozen sera from eight herds for antibodies to BLV, bovine visna virus (BVV) and bovine syncytial virus (BSV). Serum samples from 235 animals in four dairy and four beef herds were analyzed. Detection methods used included indirect fluorescent antibody tests of virus-infected cell cultures (BLV, BSV, BVV) and agar gel immunodiffusion (BLV). Sera from the BLV-infected animals in the dairy herds showed the highest single (50%, 49/97) and multiple (30%, 29/97) infections compared with 5% (7/138) and less than 1% (1/138), respectively in the beef herds. Single BVV infections were not detected in the dairy herds, but 11% (11/97) of the sera contained antibodies to BVV plus BLV or BSV. Five sera from beef cattle had antibodies only to BVV and four were obtained from one herd. Only one beef serum of the 138 tested demonstrated multiple antibodies (BLV, BVV). PMID- 2549687 TI - Inhibitory effects of ribavirin alone or combined with human alpha interferon on feline infectious peritonitis virus replication in vitro. AB - The antiviral activities of ribavirin (1-beta-D-ribofuranosyl-1,2,4-triazole-3 carboxamide; virazole), either alone or in combination with recombinant human leukocyte (alpha) interferon (rHuIFN-alpha), were evaluated against feline infectious peritonitis virus (FIPV) in feline kidney-cell cultures. The 50% inhibitory dose (ID50) of ribavirin for uninfected, rapidly dividing cells was approximately 17 micrograms ml-1 whereas the ID50 for FIPV was 2.5 micrograms ml 1. The therapeutic index (TI) of ribavirin (i.e. the ratio of the minimum cell toxic dose to minimum virus-inhibitory dose) was 6.8. Although a dose-dependent inhibition of viral infectivity occurred at non-toxic doses, maximum antiviral effects (greater than or equal to 4 log10 reduction in FIPV) occurred at cytotoxic doses. When low or moderate doses of ribavirin were combined with either 10 or 100 U of rHuIFN-alpha ml-1, the resulting antiviral effects were significantly greater than the sum of the observed effects from either ribavirin or rHuIFN-alpha alone. Significant synergistic interactions with rHuIFN-alpha occurred at ribavirin doses of 1, 5, 12.5 and 25 micrograms ml-1. Synergistic combinations of rHuIFN-alpha and ribavirin produced up to an 80-fold or a 200 fold relative increase in FIPV antiviral activities compared with that produced by equivalent doses, respectively, of ribavirin or rHuIFN-alpha alone. In cell growth studies, the addition of either 10 or 100 U of rHuIFN-alpha ml-1 to test doses of ribavirin did not increase the anticellular effect observed with ribavirin alone; seemingly, the potentiation of ribavirin antiviral activity by rHuIFN-alpha was independent of any additive cytotoxic effects. Potentially, synergistic combinations of the two antiviral agents in vivo may decrease the therapeutic dose of ribavirin required for inhibition of FIPV and thus reduce drug toxicity. PMID- 2549688 TI - Endotoxin levels in milk and plasma of mastitis-affected cows measured with a chromogenic limulus test. AB - A chromogenic limulus test ("Toxicolor") was applied to cow's milk and plasma after treatment with perchloric acid to remove interfering factors. The endotoxin levels in normal cow's milk and plasma were all less than 10 pg ml-1. In acute mastitis, the milk endotoxin level averaged (1.1 +/- 0.7) X 10(3) pg ml-1 in the cases where Gram-negative bacteria were isolated, while the plasma endotoxin concentration was normal. The endotoxin levels in the quarters infected with Gram positive bacteria were all normal, both in milk and plasma. In gangrenous mastitis due to Gram-negative bacteria, the endotoxin concentration was very high in both milk [(9.3 +/- 5.3) X 10(6) pg ml-1] and plasma (85.2 +/- 68.2 pg ml-1). In similar cases due to Gram-positive bacteria, endotoxin levels were all normal, both in milk and plasma, resembling the acute mastitis due to Gram-positive bacteria. The test was considered suitable for the diagnosis of mastitis due to Gram-negative organisms and the levels of endotoxin detected would aid in assessing the prognosis. PMID- 2549689 TI - AIDS-related diseases in animals. PMID- 2549690 TI - Feline immunodeficiency virus infection. AB - Feline immunodeficiency virus (FIV) (formerly feline T-lymphotropic lentivirus or FTLV) was first isolated from a group of cats in Petaluma, California in 1986. The virus is a typical lentivirus in gross and structural morphology. It replicates preferentially but not exclusively in feline T-lymphoblastoid cells, where it causes a characteristic cytopathic effect. The major structural proteins are 10, 17 (small gag), 28 (major core), 31 (endonuclease?), 41 (transmembrane?), 52 (core precursor polyprotein), 54/62 (reverse transcriptase?), and 110/130 (major envelope) kilodaltons in size. The various proteins are antigenically distinguishable from those of other lentiviruses, although serum from EIAV infected horses will cross-react with some FIV antigens. Kittens experimentally infected with FIV manifest a transient (several days to 2 weeks) fever and neutropenia beginning 4 to 8 weeks after inoculation. This is associated with a generalized lymphadenopathy that persists for up to 9 months. Most cats recover from this initial phase of the disease and become lifelong carriers of the virus. Complete recovery does not occur to any extent in nature or in the laboratory setting. One experimentally infected cat died from a myeloproliferative disorder several months after infection. The terminal AIDS-like phase of the illness has been seen mainly in naturally infected cats. It appears a year or more following the initial infection in an unknown proportion of infected animals. FIV has been identified in cats from all parts of the world. It is most prevalent in high density populations of free roaming cats (feral and pet), and is very uncommon in closed purebred catteries. Male cats are twice as likely to become infected as females. Older male cats adopted as feral or stray animals are at the highest risk of infection, therefore. The infection rate among freely roaming cats rises throughout life, and reaches levels ranging from less than 1% to 12% or more depending on the area. Clinically affected cats tend to be 5 years or older at the time of hospitalization. Experimental and seroepidemiologic studies suggest that FIV is transmitted mainly by bites. Intimate, non-traumatic contact (mutual grooming, shared use of food, water and litter pans) is inefficient in transmitting the infection. In utero and venereal transmission could not be demonstrated in laboratory settings. There is no statistical linkage between FIV and feline leukemia virus (FeLV) infections in nature. The FeLV infection rate in FIV-infected animals is the same as it is for non-FIV-infected cats.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2549691 TI - FeLV-FAIDS-induced immunodeficiency syndrome in cats. AB - Findings are reviewed, relevant to elucidation of the pathogenic, genetic and biochemical properties of a single, genetically heterogeneous isolate of feline leukemia virus (FeLV-FAIDS) shown to induce fatal immunodeficiency disease in nearly 100% of inoculated cats. Hypotheses are suggested which pertain to the mechanism of T-cell killing by this virus, and which extrapolate findings in the FeLV-FAIDS animal model to AIDS induced in humans by human immunodeficiency virus (HIV). PMID- 2549692 TI - Immunodeficiency in latent feline leukemia virus infections. AB - Challenge of naive experimental animals with a retroviral inoculum may result in one of two broad sequelae. The first is the establishment of an appropriate humoral and cellular immune response leading to a condition of immunity to subsequent infection with the retrovirus. Alternatively, the host may fail to develop a successful immune response, resulting in a chronic viremia associated with immunosuppression and ultimately death due to secondary pathogens. An alternate disease course is the establishment of a latent infection characterized by the presence of neutralizing antibody and strong cellular immune reactivity. Recent data from the feline leukemia virus (FeLV) system suggest that cats infected with this virus may develop immunosuppression in the form of persistent neutrophil dysfunction. The potential effect of this cellular dysfunction is the possible susceptibility of the host to the same opportunistic pathogens which are responsible for the increased mortality noted in chronic FeLV infections. These data demonstrate that persistent retroviremia is not essential for the establishment of immunosuppression. This overview presents data accumulated from the feline model of the human acquired immunodeficiency syndrome (AIDS) and discusses its relationship to human retroviral infections. PMID- 2549693 TI - FeLV-induced immunosuppression through alterations in signal transduction: changes in intracellular free calcium levels. AB - Feline leukemia virus (FeLV) is a retrovirus with immunosuppressive properties. The mechanism(s) of immunosuppression is unknown. Calcium has been shown to be a second messenger in cellular activation and regulation. This study was designed to determine whether FeLV alters intracellular free calcium (IFC) levels in an FeLV-infected feline lymphoid cell line. Control cells and FeLV-infected cells were exposed to Concanavalin A, formyl-L-methionyl-L-leucyl-L-phenylalanine, and leukotriene B4. The basal IFC and post-stimulation IFC levels were recorded using Fura 2 AM and a luminescence spectrometer. Data collected indicate that FeLV infected cells have a higher basal level of IFC and a reduced amount of increase in IFC after stimulation when compared to the control cells. The results would seem to indicate retrovirus-mediated interference occurring in the intracellular calcium signaling process. PMID- 2549694 TI - FeLV-induced immunosuppression through alterations in signal transduction: down regulation of protein kinase C. AB - Activation of protein kinase C by a phorbol ester, 12-O-tetradecanoylphorbol-13 acetate (TPA), was shown to stimulate the respiratory burst in normal cat neutrophils. Neutrophils from feline leukemia virus (FeLV)-exposed viremic and nonviremic cats had significant suppression of their respiratory burst when stimulated with TPA. The addition of whole ultraviolet light-inactivated FeLV and FeLV proteins to normal cat neutrophils produced no significant suppression of the respiratory burst. These data suggest two possible mechanisms for suppression. The first is partially due to viral alterations of the neutrophil as seen in viremic cats, but, because exogenously applied FeLV or FeLV proteins had no effect on the respiratory burst, an additional mechanism is present. The second mechanism may be caused by a latent FeLV infection residing in nonviremic cat bone marrows which alters their immune system, resulting in immunosuppression. PMID- 2549695 TI - Control of feline leukaemia virus. AB - Feline leukaemia virus (FeLV) usually occurs in its natural species, the domestic cat. FeLV is also important to human individuals as a comparative model, as it may cause a variety of diseases, some malignant and some benign, such as immunosuppression, which bears a resemblance to AIDS (acquired immune deficiency syndrome) in man. FeLV is transmitted among cats by contagion. The main sources of infection are persistently infected carrier cats which continuously excrete virus. Dissemination of FeLV among cats may be prevented by identifying infected carrier cats and removing them from contact with non-infected cats. Removal programmes using indirect immunofluorescence antibody tests were applied successfully in The Netherlands. The proportion of FeLV-positive cats decreased from 9% in 1974 to approximately 3% in 1985 during such a programme. The results of a removal programme carried out in a catbreeders' society were even better: the incidence of cats positive for FeLV decreased from 11% in 1974 to less than 2% within 4 years. None of the cats tested in this society has been found to be positive for FeLV since 1984. Besides removal programmes, other methods of control, such as pre-exposure treatment, were developed to prevent the spread of FeLV. We attempted to protect kittens against oronasal infection with FeLV by treatment with virus-neutralizing (VN) monoclonal antibodies (MoAbs) directed against an epitope on the viral glycoprotein gp70. However, no protection was achieved. It is unlikely that the amount of VN antibodies, the mode and route of their application or the infectious dose of FeLV used can account for this failure. Other possible explanations for the lack of protective effect are that (i) the restricted epitope specificity of the MoAb preparation used may have led to selection of neutralization-resistant virus mutants, or (ii) other mechanisms than virus neutralization (complement-mediated lysis, antibody-dependent cell cytotoxicity), that may be involved in protection, function less efficiently with MoAb. However, in the light of our finding that an early anti-idiotypic response is observed in all cats following administration of the MoAb preparation, the rapid clearance of anti-FeLV MoAb from the circulation is a more likely explanation. Efforts were further made to develop a vaccine for controlling FeLV infection. The immunostimulating complex vaccine (FeLV-ISCOM vaccine), a subunit vaccine in which FeLV gp70 is presented in a particular manner, looks promising. The protective effect of FeLV-ISCOM vaccine was studied by vaccinating six 8-week old SPF cats with ISCOM, followed by oronasal challenge with FeLV.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2549696 TI - Diseases associated with spontaneous feline leukemia virus (FeLV) infection in cats. AB - More than 2000 cats sent for necropsy in order to provide a diagnosis were investigated immunohistologically using paraffin sections for the presence of a persistent infection with feline leukemia virus (FeLV). The spectrum of neoplastic and non-neoplastic diseases associated significantly with FeLV infection was determined statistically. Three-quarters of the cats with persistent FeLV infections died of non-neoplastic diseases and about 23% died of tumors, nearly exclusively those of the leukemia/lymphoma disease complex. A strong association with liver degeneration, icterus and a FeLV-associated enteritis was found in addition to the known association with non-neoplastic diseases and conditions such as anemia, bacterial secondary infections and respiratory tract inflammations due to the immunosuppressive effect of FeLV, hemorrhages and feline infectious peritonitis. Surprisingly, diseases and conditions like feline infectious panleukopenia, enteritis (of other types than FeLV-associated enteritis and feline infectious panleukopenia), glomerulonephritis, uremia and hemorrhagic cystitis were not associated with persistent FeLV infection. Another unexpected finding was that most pathogenic infectious agents demonstrated in the cats were not FeLV-associated either. Thus, immunosuppression due to FeLV infection seems to make the animals susceptible to certain pathogenic infectious agents, but not to the majority. PMID- 2549697 TI - Biological variation in random and leukotriene B4-directed migration of canine neutrophils. AB - Seasonal, sexual, parental and age-related effects on peripheral blood neutrophil migration were studied in 25 dogs. Random and chemotactic movements were estimated by measuring migration towards buffer and leukotriene B4, respectively. Significant effects were seen only in the comparison between the two sexes. Neutrophils from bitches exhibited 19% greater random migration but 10% smaller chemotactic responsiveness than neutrophils from male dogs. A progesterone mediated suppression of chemotaxis is hypothesized in metoestral bitches. It is concluded that the observed differences are probably too subtle to play any role in combatting bacterial infections, but may constitute a source of bias in the evaluation of isolated cases of disease-associated defective chemotaxis. PMID- 2549698 TI - Milk whey as a serum supplement for the growth of FMD virus in BHK-21 cells. AB - Two batches of experimental media containing 10% milk whey (sweet or sour) and 1% bovine serum supported the growth of BHK-21 cells in serial passages. The cell yield was greater than 2 x 10(6)/ml at 48 h. The cells were susceptible to inoculation with foot and mouth disease (FMD) virus type 'A' and the virus titres obtained were comparable with those in Eagle's MEM-G with 10% serum. The use of milk whey could reduce the consumption of serum required for the growth of FMD virus in BHK-21 cells by up to 90%. PMID- 2549699 TI - Epstein-Barr virus (EBV). V. Incidence of EBV antibodies in patients with rheumatic diseases. AB - Serum samples from 95 patients with rheumatoid arthritis, 24 patients with other various rheumatic diseases, 50 patients with diabetes mellitus, 34 patients with acute viral infections, 6 patients with infectious mononucleosis, 77 patients with lymphomas and leukemia and 110 blood donors and 24 healthy subjects as normal controls, respectively, were tested by indirect immunofluorescence (IF) reaction for the presence of specific antibodies against Epstein-Barr virus determined viral capsid antigen (anti-VCA) and Epstein-Barr active viral infection. The IF test carried out in acetone-fixed smears of EB-3 cell line revealed EB antibodies anti-VCA in 83.3% of infectious mononucleosis, 61.0% lymphomas and leukemia, 58.0% diabetic patients. The frequency of anti-VCA antibodies in rheumatic patients was 31.4%, and 3.6% and 25% in sera from blood donors and healthy subjects, respectively. Incidence of active EBV infection was 5.7% of rheumatic diseases, 17.7% of acute virus infections, 50.0% of infectious mononucleosis, and 31.1% of lymphomas and leukemia patients. Active EBV infection was not found out in blood donors (0/110) and healthy subjects (0/24) groups as control. Rheumatoid arthritis with or without rheumatoid factor patients had serological evidence of active EBV infection 6/26 and 4/26 respectively. PMID- 2549700 TI - Epstein-Barr virus (EBV). VI. Incidence of EBV antibodies in patients with malignant lymphoproliferative diseases. AB - Serum samples from 52 patients with malignant lymphoproliferative diseases and 12 clinically healthy subjects were tested by indirect immunofluorescence (IF) reaction for the presence of specific EB antibodies anti-VCA and active EBV infection. The tests revealed EB antibodies anti-VCA in 32 patients with lymphoproliferative diseases and in 2 clinically healthy subjects and active EBV infection in 18/32 and in 0/2 EB anti-VCA positive patients and clinically healthy subjects, respectively. PMID- 2549701 TI - Epstein-Barr virus (EBV). VII. Established lymphoid cell line (IVPat-88) obtained from synovial fluid of a patient with aseptic arthritis. AB - Attempts have been made to culture mononuclear cells from synovial fluid of 8 patients with arthropathy, and have led to the development of the lymphoid cell line IVPat-88. Cell line has been propagated by serial passages for more than 14 weeks in continuous culture. The cells grew as single, free-floating individuals, or in dense clumps without adherence to glass or plastic surface. All these cells were identified as altered lymphoblasts because of their growth pattern and uniform morphology, and the presence of Epstein-Barr Viral Capsid Antigen (VCA) in 5 to 10% of the cells. The cell concentration varied during the period of culture from about 300,000 to 1,700,000 cells per ml, and mean doubling time during phases of active growth was 42 and 60 hours in MEM and RPMI 1640 tissue culture media, respectively. The methods used and the characteristics of the cell line are described. PMID- 2549702 TI - [Virologic research on the quality of 2 sources of drinking water from zone "I"]. AB - Enteroviruses were isolated from 4.6% out of the examined tap water samples. No relation could be established between viral pollution and classic quality indexes of water. PMID- 2549703 TI - [Adaptation of the avidin-biotin system for identifying and quantifying Sendai virus antigens]. AB - The avidin-biotin system was adapted in view of the identification and dosage of the Sendai parainfluenza virus and of its antigens, using the method of double antibodies (biotinylated and nonbiotinylated) in ELISA type tests. PMID- 2549704 TI - [The protector effect of ribosomal preparations against experimental influenza infection in mice]. AB - A study was conducted on the protective effect of some ribosomal preparations, isolated from chorionic-allantoic membranes of chicken embryos, infected or not with parainfluenza (Sendai) or influenza (AoPR8) virus, in mice experimentally inoculated with influenza virus strain AoPR8 adapted to the mouse. Results showed that the tested preparation, containing ribosomes and polysomes isolated from chorio-allantoic membranes of Sendai virus inoculated chicken embryos, ensure the mice complete protection against AoPR8 virus, if administrated before the control infection. PMID- 2549705 TI - Viral transcripts in cells infected with defective interfering particles of equine herpesvirus type 1. AB - Equine herpesvirus type 1 (EHV-1) preparations enriched in defective interfering particles (DIPs) have previously been demonstrated to mediate the coestablishment of persistent infection and oncogenic transformation in primary hamster embryo fibroblasts. In this study, it was demonstrated that infection of a rabbit kidney (RK) cell line with EHV-1 DIP-enriched preparations also results in the establishment of persistent infection. Viral transcription was characterized in RK cells infected with DIP-enriched stocks and compared to viral transcription in RK cells infected with standard (STD) EHV-1. During the first 8 hr of infection with the DIP-enriched EHV-1 preparation, viral DNA sequences which are conserved in the DIP genome were predominantly expressed. Thus, these transcripts originate from DNA sequences that contain the components of the defective genome which originates from DNA sequences mapping at 0.00-0.04 of the Long region terminus and within two portions of the Short region inverted repeats (IR), 0.78-0.79 and 0.83-0.865 of the internal IRs and 0.99-1.00 and 0.915-0.95 of the terminal IRs. The overwhelming majority of viral transcripts that were synthesized in the DIP enriched infections appeared to correspond to transcripts expressed in STD infection as assessed by Northern hybridization analysis but the synthesis of transcripts originating from sequences not conserved in the defective genome was significantly delayed. However, some high molecular weight RNA species that were synthesized in STD infections were not detected in DIP-enriched infections. Studies utilizing metabolic inhibitors indicated that viral transcription in DIP enriched infections, like that of STD cytocidal infection, is regulated in an immediate early, early and late manner. PMID- 2549706 TI - Generation of a nucleosome-free promoter region in SV40 does not require T antigen binding to site I. AB - The relationship between T-antigen interactions at Site I and the presence of a nucleosome-free promoter in SV40 chromatin was examined by analyzing chromatin from mutants defective for T-antigen interaction at site I (cs 1085, scs111, and tsA58) and their parental wild-type strains (776, SVS, and VA45-54, respectively). As judged by sensitivity to digestion with restriction endonucleases that recognize unique sequences within the promoter region (BglI, KpnI, and MspI), a nucleosome-free promoter was observed in a substantially larger proportion of chromosomes from the defective mutants than their wild-type parents. This result demonstrates that T-antigen binding to site I is not necessary for setting the early boundary of the nucleosome-free region, although it may function directly or indirectly in determining the proportion of chromosomes containing this feature. PMID- 2549707 TI - Transposon mutagenesis of baculoviruses: analysis of Trichoplusia ni transposon IFP2 insertions within the FP-locus of nuclear polyhedrosis viruses. AB - The transposable IFP2 element of Trichoplusia ni was originally isolated as a host DNA insertion in spontaneous FP mutants of Galleria mellonella or Autographa californica nuclear polyhedrosis viruses (NPVs). The termini of IFP2 insertions from five independently isolated FP mutants were sequenced. In all cases IFP2 is flanked by 13-bp terminal inverted repeats and has additional inverted repeats of 19 bp in length located asymmetrically with respect to the ends of the element. Insertion of IFP2 into the viral genome always generated a duplication of the tetranucleotide target site, TTAA. There was an apparent preference for insertion within a 12-bp A + T-rich imperfect palindromic sequence surrounding the target site. Sequence analysis of three independent IFP2 elements revealed an internal domain of 2.475 kb containing an RNA polymerase II promoter region and two large open reading frames. Primer extension analysis of IFP2-specific mRNA positioned the 5' terminus of the transcript. The element is present in DNA isolated from T. ni cell lines TN-368 and TN-5B1, but is not apparent in DNAs isolated from the TN R2 cell line or our laboratory colony of T. ni larvae, suggesting IFP2 was recently introduced into the T. ni genome. PMID- 2549708 TI - Papillomavirus-associated inductions of cellular proteins in mouse C127 cells: correlation with the presence of open reading frame E2. AB - Bovine papillomavirus type 1 (BPV-1) readily transforms mouse C127 cells, conferring the ability to grow in soft agar and to form tumors in athymic (nu/nu) mice. Electrophoresis of total cellular proteins from these BPV-transformed lines on ultra-high resolution, giant two-dimensional gels displays the presence of novel, papillomavirus-related protein phenotypes. Analysis of the established BPV 1-transformed C127 cell lines, ID13 and ID14, reveals a set of six proteins which are either absent or synthesized at extremely low levels in the parental cell line. One of these proteins is also present in v-ras-transformed C127 cells, but none of the others are found in cells transformed by a variety of viral oncogenes, including the polyomavirus middle T, v-mos, or v-fes. The genome of BPV-1 contains two separate open reading frames (ORFs), E5 and E6, which can act independently to transform C127 cells. In addition, trans-activator and repressor proteins encoded respectively by the full-length and carboxy-terminal E2 ORF regulate the level of expression of other BPV-1 genes. We examined 34 cell lines transformed by intact and subgenomic recombinant DNAs of BPV-1. Cells harboring BPV-1 DNAs engineered to eliminate the expression of ORFs E4, E5, E6, or E7 display five of the PV-associated proteins, but these proteins are not seen in lines lacking the full E2 ORF. Moreover, G418-selected nontransformed cells expressing E2 cDNA from an SV40 promoter exhibit these proteins at high levels. Surprisingly, these proteins are also present in cells containing BPV-1 DNAs with amino-terminal E2 deletions, suggesting that these PV-associated proteins represent novel cellular responses to a factor encoded within the E2-C gene region. PMID- 2549709 TI - Sequence analyses and structural predictions of double-stranded RNA segment S1 and VP7 from United States prototype bluetongue virus serotypes 13 and 10. AB - The nucleotide sequence of segment S1 and the deduced amino acid sequence of VP7 from bluetongue virus (BTV) serotype 13 was determined. Sequences were obtained by use of standard dideoxy DNA sequencing and by direct sequencing of genomic double-stranded RNA (dsRNA). The dsRNA was sequenced with a new dideoxy protocol that produces 300 to 350 bases per set of reactions. Segment S1 is 1156 bp long and contains one long open reading frame capable of coding for 349 amino acids. The protein, VP7, is rather hydrophobic, and has a calculated molecular weight of 38,619 and a net charge of +1.5 at pH 7.0. Segment S1 of BTV-13 has 79.6% of its nucleotides conserved when compared with segment S1 of BTV-10. While most of these differences occur at the third codon position of the open reading frame, the differences between the 89-base-long, 3' noncoding regions occur predominantly in pockets at positions 1092-1098, 1112-1114, and 1125-1129. Potential stem-loop structures encompassing the stop codon of the open reading frame are proposed for both serotypes. Comparisons of VP7 from BTV-13 and BTV-10 indicate that 93.7% of the amino acid residues are conserved, including a single lysine at position 255. Secondary structure predictions infer an eight-stranded beta-barrel structure between residues 150 and 250. This putative beta-barrel may serve as a target for the development of drugs to combat bluetongue disease. Comparable structures detected in the core proteins of single-stranded RNA viruses from both plants and animals suggest that these viruses and BTV had a common origin. PMID- 2549710 TI - Comparison of human, simian, and bovine rotaviruses for requirement of sialic acid in hemagglutination and cell adsorption. AB - Human rotaviruses (Wa, KUN, MO) showed hemagglutination (HA) only with fixed 1 day-old chicken erythrocytes, and their HA activities were completely destroyed by trypsin activation of virions. Simian SA-11 and bovine NCDV had HA activities not only against fixed erythrocytes but also against fresh erythrocytes from various species. Their HA activities against fixed erythrocytes were also inhibited by trypsin activation, but those against fresh erythrocytes were not. Neuraminidase treatment of fixed erythrocytes did not inhibit HA by trypsin untreated rotaviruses. In contrast, HA of fresh human erythrocytes by SA-11 and NCDV was completely inhibited by neuraminidase treatment of erythrocytes or glycophorin A, the major erythrocyte sialoglycoprotein. Adsorption and infection of SA-11 and NCDV to monkey kidney MA104 cells were also inhibited by neuraminidase treatment of cells. Adsorption and infection of human rotaviruses were not, however, affected by treatment of cells with neuraminidase from Vibrio cholerae or Arthrobacter ureafaciens or with potassium periodate. Therefore, HA of fixed chicken erythrocytes by trypsin-untreated human and animal rotaviruses may be independent of sialic acids, whereas that of fresh erythrocytes by SA-11 and NCDV is sialic acid dependent and probably mediated by glycophorin A. Sialic acids also constitute an essential part of the cellular receptors for SA-11 and NCDV, whereas those for human rotaviruses were quite resistant to treatments known to destroy major types of sialic acids. PMID- 2549711 TI - DNA sequence and comparative analyses of the equine herpesvirus type 1 immediate early gene. AB - The immediate early (IE) proteins of herpesviruses are important regulatory factors which control the expression of genes at the transcriptional level. We report the DNA sequence of the immediate early gene of the alphaherpesvirus equine herpesvirus type 1 (EHV-1). This sequence is shown to be extremely rich in guanine and cytosine, resulting in a highly biased codon usage. The IE gene region possesses 38 open reading frames (ORFs) greater than 300 bp in length, 11 of which have coding regions of at least 100 amino acids (aa) following potential translation initiator codons. The largest ORF consists of 1487 codons (4461 bp) starting with the first ATG and would encode a protein of MW 155,000. TATA and CCAAT sequences as well as several potential cis-acting elements lie upstream to the major ORF. The deduced amino acid sequence for the 155,000 protein has a high degree of homology to the herpes simplex virus type 1 (HSV-1) ICP4 protein and its varicella-zoster virus (VZV) homolog. The regions of the EHV-1 IE protein that are homologous with these proteins correspond to the previously determined pattern of homology between the HSV and VZV IE polypeptides. However, there are are a number of differences within these broadly defined regions. It is therefore expected that this comparative study will facilitate the identification of functionally important residues within the amino acid sequence of IE proteins. PMID- 2549712 TI - Tumorigenic poxviruses: characterization of an early promoter from Shope fibroma virus. AB - A strong early promoter from the T1 open reading frame (ORF) within the terminal inverted repeat (TIR) of Shope fibroma virus (SFV) has been isolated and characterized. Promoter activity was determined by a transient gene expression assay in poxvirus-infected cells using the bacterial chloramphenicol acetyltransferase as a reporter gene. The sequences which constitute the boundaries of the promoter element were determined by 5' and 3' deletion analysis. The functional SFV T1 promoter domain comprises about 28 bp and includes, in addition to the transcriptional initiation site, a stretch of eight continuous A residues from position -18 to -11 which is critical for promoter function. Both the SFV T1 promoter and the vaccinia 7.5-kDa early/late promoter are active in the transient expression assay when the cells are infected with either the leporipoxvirus SFV or the orthopoxvirus vaccinia. To look more closely at the conservation of promoter function between poxvirus genera, a recombinant vaccinia virus containing the CAT gene driven by the SFV T1 promoter and a recombinant SFV containing the CAT gene driven by the vaccinia 7.5-kDa early/late promoters was constructed. The SFV T1 promoter behaves as an early promoter in the vaccinia genome, and both the T1 and the 7.5-kDa early/late promoters use transcriptional initiation sites in their heterologous genomic environment that are identical to the ones used in the native viral genome. The results from this work indicate that despite the relative lack of absolute sequence conservation, the transcriptional machinery, at least with respect to temporal regulation of early promoters and the position of transcript initiation, is conserved between these two poxvirus genera. PMID- 2549714 TI - The adeno-associated virus Rep78 gene inhibits cellular transformation induced by bovine papillomavirus. AB - Adeno-associated virus type 2 (AAV) is a helper dependent parvovirus which can inhibit the oncogenic and transforming potential of its helper viruses: adenoviruses (Ad) and herpesviruses. Here we have assayed AAV's ability to inhibit cellular transformation induced by bovine papillomavirus type 1 (BPV-1), a member of another family of DNA viruses. AAV was able to markedly inhibit BPV-1 induced transformation of contact-inhibited murine fibroblasts either by infection with virus particles or by DNA transfection. This inhibition was mapped to the full-length, unspliced, rep gene product, Rep78, of AAV. DNA replication by AAV or by BPV-1 was not required for inhibition of transformation to take place. PMID- 2549713 TI - In vitro infection of woodchuck hepatocytes with woodchuck hepatitis virus and ground squirrel hepatitis virus. AB - Primary cultures of woodchuck hepatocytes were demonstrated to be susceptible to in vitro infection by both woodchuck hepatitis virus and ground squirrel hepatitis virus, as evidenced by the appearance of DNA species characteristic of hepadnavirus replication. Initiation of infection by woodchuck hepatitis virus was blocked by the presence of suramin, polybrene, or dideoxycytidine. Viral CCC DNA, the putative template for viral RNA transcription, was detected at 2 days postinfection. Accumulation of intracellular intermediates in virion DNA synthesis was negligible until 7-10 days postinfection, but these DNA intermediates then increased dramatically in amount over the next few weeks. Results were obtained which suggested that the prolonged accumulation of intermediates in virion DNA synthesis was an intrinsic property of the infection of individual cells, and not the result of a slow spread of virus through the cultures. PMID- 2549715 TI - Inhibitory effects of vesicular stomatitis virus on cellular and influenza viral RNA metabolism and protein synthesis. AB - Infection with vesicular stomatitis virus (VSV) results in the rapid inhibition of cellular macromolecular synthesis, including transcription, translation, and maturation of the U1 and U2 snRNPs. Unlike infection with VSV, influenza virus infection did not result in the inhibition of either the processing of U1 and U2 snRNAs or the assembly of the RNPs. Although influenza virus relies on the cellular splicing apparatus to generate viral mRNAs, the maturation of snRNPs was inhibited during double infections with VSV. However, this inhibition of snRNP maturation had no effect on the splicing of a cellular pre mRNA in extracts prepared from VSV-infected HeLa cells. Thus, the effects of VSV on the processing and assembly of snRNPs appear to involve virus-specific functions and unique cellular targets. Coinfection with VSV and influenza virus resulted in the dramatic inhibition of influenza virus transcription; polyadenylated mRNAs corresponding to the influenza virus NP and NS1 proteins could not be detected by Northern blot analysis. However, reduced levels of the influenza virus replicative templates were still synthesized during double infection. Coinfection with VSV also resulted in the inhibition of influenza viral mRNA translation, even when superinfection with VSV was delayed until 3 or 6 hr after influenza virus infection. VSV required at least 2 hr to affect the inhibition of translation; this corresponded to the time after VSV infection when inhibition of cellular protein synthesis was evident. These results demonstrate that, in contrast to adenovirus, the VSV-mediated inhibition of cellular macromolecular synthesis may be effective against influenza virus. PMID- 2549716 TI - Differentiation-linked human papillomavirus types 6 and 11 transcription in genital condylomata revealed by in situ hybridization with message-specific RNA probes. AB - Human papillomaviruses (HPVs) infect specific human epithelial tissues. Because viral propagation in cultured cells has not been achieved, studies of HPV genetic activities have been difficult and rely largely on analyses of patient specimens by conventional biochemical methods. HPV type 6 and type 11 infections often result in genital warts (condylomata acuminata). Structural mapping of RNAs from such warts reveals that they use alternative promoters, splice sites, and polyadenylation sites to produce complex families of overlapping mRNAs that span multiple open reading frames. Based on the mRNA structures, we have developed message-specific subgenomic clones of HPV-6 and HPV-11 in pGEM vectors. Tritium labeled, single-stranded RNA probes were synthesized in vitro and applied to serial thin sections of patient specimens for in situ hybridization. Our results reveal the qualitative and quantitative transcription patterns of different viral messages in relationship to one another, to viral DNA replication, and to cellular differentiation. The viral "E region" is transcribed before the onset of vegetative DNA replication and continues to be expressed in increasing amounts in the maturing epithelium. Even in mature epithelia, E region messengers are far more abundant than "L region" mRNAs. The L region messages encoding capsid proteins are truly late in that they appear concomitant with or after the onset of vegetative viral DNA replication and are only present in the superficial strata of the epithelium, which contain the oldest and most differentiated keratinocytes. Abundant intron material derived from processing E region transcripts accumulates in the nuclei. Strictly nuclear signals from the L region transcripts in the midepithelium suggest that regulation of their expression is at the level of transcription elongation. PMID- 2549717 TI - Replication of the genome RNAs of defective interfering particles of vesicular stomatitis and Sendai viruses using heterologous viral proteins. AB - We have tested the ability of heterologous viral proteins to support the in vivo and in vitro replication of the RNA of defective interfering (DI) particles of two serotypes of VSV and of Sendai virus. In all the combinations of heterologous coinfections in vivo, DI particle replication was observed only in the coinfection with the VSV-Indiana DI particle and wild-type VSV-New Jersey. By quantitating RNA synthesis in reconstitution experiments we showed that with DI nucleocapsids isolated from infected cells, however, the soluble protein fraction from heterologous wild-type virus-infected cells could substitute in vitro to varying degrees for the homologous proteins in the elongation reaction of RNA replication and encapsidation. In these cases successful replication was confirmed by demonstrating the specific association of the heterologous N protein with the product nucleocapsid RNA. The initiation step, that is, the initial binding of the nucleocapsid protein to the leader RNA, in contrast, requires the homologous protein, since heterologous viral proteins could not support RNA replication and encapsidation from purified DI particles. PMID- 2549718 TI - Protein analysis of herpes simplex virus latency in vitro established with cycloheximide. AB - Herpes simplex virus (HSV)-specific protein synthesis was examined during establishment of HSV latency and reactivation of virus in human embryonic lung cells treated with cycloheximide and incubated at 40.5 degrees. Eight viral proteins, identified during the first two days of establishment of latency at 40.5 degrees, were undetectable by Day 3. At least two synthesized proteins were present during the maintenance phase of latency. Reactivation of HSV (viral protein 135K) was first detected in latently infected cultures between 2 and 3 hr after superinfection with human cytomegalovirus (HCMV). During this period an 82K protein with the same molecular weight as one of the HCMV immediate-early proteins (82 and 75K) was detected in the immunoprecipitates of latently infected cultures with anti-HSV serum. Thus, this HSV latency system can be used to analyze protein synthesis and clarify reactivation of HSV by HCMV superinfection. PMID- 2549719 TI - An unusual coordinated cleavage event in the processing of encephalomyocarditis virus polypeptides. AB - During the translation of encephalomyocarditis virus RNA in the rabbit reticulocyte lysate system, polypeptides 2B and 2C, encoded by the central region of the genome, appear simultaneously in the absence of any detectable 2BC precursor and at a time when translation has advanced into the region coding for polypeptide 3C. This implies the operation of closely coordinated proteolysis at two sites, with at least one cleaved by the virus-coded protease activity 3C. PMID- 2549720 TI - Two domains distantly related to protein-tyrosine kinases in the vesicular stomatitis virus polymerase. AB - We have carried out an exhaustive search for amino acid sequence similarities between vesicular stomatitis virus (VSV) proteins and database entries. Unexpectedly, we found that the L polymerase protein contains two blocks of sequence (residues 725-1102 and 1291-1671) with distant but statistically significant similarity to the catalytic domain of tyrosine-specific protein kinases. The first kinase-like region is most similar to members of the Abl subfamily, Fes and Fps (26.6% and 27.3% identity, respectively), whereas the second region is closest to members of the platelet-derived growth factor receptor (PDGFR) subfamily, PDGFR and Kit (30.4% and 25.9% identity, respectively). Multiple alignment of the catalytic domain of these kinases to all three rhabdovirus L protein sequences available (VSV Indiana, VSV New Jersey, and rabies) revealed that the polymerases contain many but not all residues well conserved in the protein kinase family. Similarity was highest for VSV Indiana and lowest for rabies. We conclude that the kinase-like regions in the rhabdoviral L proteins are probably very distantly related to the protein kinase family. The similarities could either reflect contemporary protein kinase activity or represent some other function(s) associated with these large multifunctional polymerase proteins. Our findings also shed new light on questions of the origins and evolution of RNA viruses. PMID- 2549721 TI - The role of monovalent cation transport in Sindbis virus maturation and release. AB - Alterations in intracellular monovalent cation concentrations in Sindbis virus infected avian cells result, in part, from a reduction in Na+/K+ ATPase (Na+ pump) activity. Inhibition of Na+ pump activity was shown previously to temporally correlate with the appearance of viral envelope proteins on the cell surface and the release of virus particles. Cells infected with envelope defective temperature-sensitive mutants exhibited reduced Na+ pump activity at the nonpermissive temperature, where viral particles are not released. By contrast, Na+ pump activity was not inhibited in Sindbis virus-infected cells treated with tunicamycin or with antiviral serum, which block virus maturation and release. Diuretic-sensitive transport of 86Rb+, aK+ tracer, was stimulated in cells which express virus envelope proteins, but fail to release virus particles. In these cells, the furosemide-sensitive 86Rb+ influx exhibited an increase in Vmax and was responsive to changes in the extracellular concentration of NaCl. Furosemide inhibited the rapid release of virus from low salt-inhibited cells after shift to isotonic conditions. Alterations in ion transport during alphavirus infection may, therefore, facilitate the efficient release of progeny virus particles. PMID- 2549722 TI - Sequence divergence yet conserved physical characteristics among the E4 proteins of cutaneous human papillomaviruses. AB - Human papillomavirus (HPV) types 1, 2, and 4 together comprise the major cause of cutaneous papillomas in the general population. We have aligned the genomes of these three viruses by partial sequence analysis, and have sequenced the E4 open reading frames (ORFs) of HPV 2 and HPV 4. After expression as beta-gal fusion proteins in bacteria, antibodies raised to the putative E4 gene-products of both virus types were used to identify the native E4 proteins in naturally occurring tumors. At the primary amino acid sequence level, the E4 protein of HPV 2 was found to be most homologous with those of HPV 6 and 11 and was not closely related to those of HPV 1 or 4. Although the E4 ORF represents a region of weak homology amongst papillomaviruses, the E4 encoded proteins showed significant conservation in their physical characteristics. Like those of HPV 1, the E4 proteins of both HPV 2 and HPV 4 were found to be composed of a major low molecular-weight doublet (16.5/18K for HPV 2, 20/21K for HPV 4, c.f. 16/17K for HPV 1) along with minor high-molecular-weight species, which probably represent dimers of the smaller proteins, (33K for HPV 2, 40K for HPV 4, c.f. 32/34K for HPV 1). The E4 products of all three virus types were multiply charged, and exhibited a characteristic migration pattern following alkaline urea gel electrophoresis. Although the levels of E4 expression in tumors induced by the different virus types was very different, this was found to correlate closely with the level of virus production characteristic of each virus type. In all three cases, E4 proteins were found to be primarily cytoplasmic, and to be associated with the distinctive cytoplasmic inclusion granules characteristic of each virus type. The poor sequence conservation between the E4 protein of HPVs 1, 2, and 4, taken alongside the ability of these viruses to infect similar histological sites, suggests that E4 may not be involved in determining tissue specificity. Our results suggest conserved physical characteristics (acidic, multiply charged, ability to form dimers) and similar site of expression may be the important factors for E4 function. PMID- 2549723 TI - Both episomal and integrated forms of human papillomavirus type 16 are involved in invasive cervical cancers. AB - The form of human papillomavirus type 16 (HPV 16)DNA in specimens of invasive cervical cancer was investigated. High molecular, tandem repeats of viral sequences were detected as several distinct bands, using a low concentration (0.5%) agarose gel and a no-cut enzyme (HindIII) for HPV 16. Two-dimensional agarose gel electrophoresis allowed us to differentiate between the episomal multimeric and the integrated forms of viral DNA. All 34 cervical cancer specimens showed the characteristic PstI cleavage pattern of HPV 16 DNA, indicating that a full viral genome was present in these specimens, and 24 specimens (70%) showed only episomal monomeric or multimeric forms without the integrated form of HPV 16 DNA. The remaining 10 specimens (30%) showed integrated multimeric forms of viral DNA, either without the episomal form (8 specimens) or with the concomitant episomal form (2 specimens). In addition, a metastatic tumor in a pelvic lymph node showed only the episomal form of viral DNA, whereas its primary cervical cancer showed both episomal and integrated forms of viral DNA. There was no correlation between the forms of viral DNA and the clinical stages of tumors. The result indicates that both episomal and integrated forms of a complete HPV 16 DNA are involved in invasive cervical cancers. PMID- 2549724 TI - Amplification of bovine papillomavirus DNA by N-methyl-N'-nitro-N nitrosoguanidine, ultraviolet irradiation, or infection with herpes simplex virus. AB - Treatment with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or irradiation with ultraviolet light (uv254 nm) induces amplification of integrated as well as episomal sequences of bovine papillomavirus (BPV) type 1 DNA in BPV-1-transformed mouse C127 cells (i.e., ID13 cells). This is shown by filter in situ hybridization and Southern blot analysis of cellular DNA. Similarly, infection of ID13 cells with herpes simplex virus (HSV) type 1 which has been shown to be mutagenic for host cell DNA leads to amplification of BPV DNA sequences. In contrast to this induction of DNA amplification by initiators, treatment of ID13 cells with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) does not result in increased synthesis of BPV DNA nor does TPA treatment modulate the initiator-induced DNA amplification. Similar to other cell systems infection with adeno-associated virus (AAV) type 2 inhibits BPV-1 DNA amplification irrespective of the inducing agent. In contrast to initiator-induced DNA amplification, treatment with carcinogen (MNNG) or tumor promoters or combination of MNNG and promoter of C127 cells prior to transformation by BPV-1 does not lead to an increase in the number of transformed foci. The induction of amplification of papillomavirus DNA by initiating agents possibly represents one of the mechanisms by which the observed synergism between papillomavirus infection and initiators in tumorigenesis might occur. PMID- 2549725 TI - Activation of HIV LTR-directed expression: analysis with pseudorabies virus immediate early gene. AB - The long terminal repeat (LTR) region of the human immunodeficiency virus (HIV 1), which regulates viral gene expression, is modulated by viral trans-acting proteins of HIV and DNA viruses and by biologically active chemical agents that induce cellular proliferation and/or differentiation. The pseudorabies virus immediate early gene (PIE) shares similar transcriptional trans-activating properties with the gene products of several other DNA viruses. The transient expression chloramphenicol acetyl transferase (CAT) assays in HeLa cells transfected with HIV long terminal repeat (LTR)-CAT and PIE plasmids demonstrated trans-activation of the HIV LTR by PIE. Analyses of 5' deletion mutants and site directed Sp1 and transactivation responsive (TAR) region mutants of the LTR indicated PIE-responsive sequences located between -65 and -17. Synergistic cooperativity between PIE and the HIV-1 tat protein was demonstrated. PIE exhibited a marked stimulatory effect upon HIV replication in HeLa cells transfected with a biologically active HIV proviral DNA. These data provide evidence that, like a number of other DNA containing viruses, PRV can trans activate HIV gene expression. PMID- 2549726 TI - [Various biochemical indicators in prenatally alcoholized rats]. AB - A number of biochemical parameters have been determined which characterized the state of CNS and the structure of cell membranes in antenatally alcoholized animals. As compared with the control group the alcoholized animals had a significantly increased level of phosphatidyl inositol diphosphate in brain phospholipid fractions, while content of serotonin was reduced in their brain and increased in blood plasma; content of phosphatidyl serine and the cholesterol/phospholipids ratio were reduced in brain. PMID- 2549727 TI - [Lipid peroxidation, the enzyme antioxidative system and acid phosphatase content of the gastric mucosa in stomach ulcer]. AB - Lipid peroxidation, a state of the antioxidative system and activity of acid phosphatase were studied in mucosal membrane of 53 patients with ulcerous disease of stomach. Increase of the acid phosphatase activity in cytoplasm, activation of lipid peroxidation and inhibition of the antioxidative system were detected in ulcer border and periulcerous region. These alterations in lipid peroxidation and the state of the antioxidative system were considered as distinct pathogenetic factors responsible for deterioration of lysosomal membranes and contributing to chronic and relapsing development of ulcerous disease. PMID- 2549728 TI - [Catecholamine complex-adrenoreactive system of various subtypes in various forms of bronchial asthma in children]. AB - During the acute period of diverse forms of bronchial asthma in children the following alterations were detected in catecholamine metabolism and in various subtypes of the adrenoreactive system: an increase in urinary excretion of catecholamines, especially of adrenaline, as well as activation of catechol-O methyl transferase; a decrease (or even absence) of beta 2-adrenoreactive system sensitivity, a decrease in beta 1-reactivity, while sensitivity of alpha 1 adrenoreactive system was increased (in the infectious-allergic form of the disease) and alpha 2-reactivity was unaltered. The uncoupling between catecholamines and various subtypes of the adrenoreactive system appears to occur during the acute period of bronchial asthma. PMID- 2549729 TI - [The effect of a controlled physical load on the level of cyclic adenosine monophosphate in plasma and thrombocytes]. AB - The authors studied the functional activity of thrombocytes, levels of plasma and thrombocytic cyclic AMP (cAMP) in the dynamics of a veloergometric test in patients with IHD and neurocirculatory dystonia. The revealed signs of hyperactivation of thrombocytes and absence of compensatory response of cAMP in IHD patients against the background of thresholds loads allows to improve the quality of prediction of thrombotic complications and control of treatment efficacy with agents possessing antithrombocytic activity. PMID- 2549731 TI - [Tumor markers in bronchus cancer]. AB - Small cell lung cancers are neuroendocrine tumours and therefore produce a lot of peptide hormones (calcitonin, ACTH, ADH), as well as the neuropeptide chromogranin A, which are all useful tumour markers. Furthermore, the tumour associated antigens CEA and TPA, as well as the enzymes neuron specific enolase (NSE) and creatine kinase BB are used as markers in small cell lung cancer. At present, NSE appears to be the best marker for small cell lung cancer; elevated serum NSE levels are found in 65 to 85% of the patients. The serum level of the tumour markers is related to the stage of the tumour. When tumour regression occurs following therapy, elevated pretreatment levels decrease to the normal range. If the marker level increases again, tumour progression is indicated and this can be an early and sensitive sign denoting recurrence. Metastases in the central nervous system can be detected early by marker determination in the cerebrospinal fluid. At present, CEA appears to be the most valuable tumour marker for non-small cell lung cancer, but TPA may also be a useful marker. PMID- 2549730 TI - Diurnal rhythms of cortisol, ACTH, and beta-endorphin levels in neonates and adults. AB - To determine whether a diurnal rhythm exists in neonates admitted to neonatal intensive care units where there is continuous artificial lighting and periodic nursing and medical care, plasma cortisol, adrenocorticotropin (ACTH), and beta endorphin concentrations were measured in two groups of infants and in adult human volunteers. As expected, a diurnal rhythm was seen in adults. A diurnal rhythm was also found for cortisol and endorphin levels in neonates (3 to 4 days postnatally) with minimal stress and in infants who were clinically severely stressed. There was not a significant difference between the morning and afternoon concentrations of ACTH in these infants, but the afternoon concentrations were lower than the morning's, as would be expected. We found that a diurnal rhythm does exist in neonates within the first few days of postnatal life and that the continuous lighting and medical and nursing interventions do not interfere with this rhythm. PMID- 2549732 TI - [Tumor markers in internal medicine hepatology]. AB - Primary liver tumour, i.e. hepatocellular carcinoma (HCC) is one of the world's most common malignancies. The age-standardized incidence rate varies widely from 6/100,000 (Austria) to 100 and more per 100,000 in Mozambique and Taiwan. In these high-risk areas, infection with hepatitis B virus is considered the main risk factor. In low-incidence areas as in Western Europe, main risk factors are older age, male sex and liver cirrhosis. The prognosis depends mainly on the size of the tumour. Alpha-fetoprotein is an ideal tumour marker for prospective screening in high-incidence areas. There is not much information, however, on the value of this marker for screening in low-incidence areas. There is some information that HCC-associated alkaline phosphatase could be a good tumour marker in non-viral liver tumours. Furthermore, des-gamma-carboxyprothrombin and vitamin B 12-binding protein could be specific markers for tumours in non cirrhotic livers. But large clinical studies have not yet confirmed the value of these markers. Hormonal and haematological markers and some isoenzymes are tumour markers characterized by high sensitivity but low specificity. There are no specific tumour markers for metastatic liver disease. PMID- 2549733 TI - [Retroviruses and their importance in neurology]. AB - Retroviruses comprise spumaviruses, lentiviruses and oncoviruses. After these taxonomic remarks neurologic-psychiatric-syndromes (multifocal leucencephalopathy, dementia-complex, vacuolar myelopathy and others) are described, that could be observed as manifestations of the "acquired immune deficiency syndrome". Some therapeutic measures are discussed, especially the administration of Zidovudine that shows good temporary results. Essentially Zidovudine works by misleading the Reserve Transkriptase by offering a "Fake thymidine"--so misleading replication. Lastly the hypotheses concerning multiple sclerosis are put in relation to lentiviruses. Some animal variants of the viruses and their neurotropic features are described. This discovery has lead to those speculations--however, these speculations remain unsecure. PMID- 2549734 TI - Urinary metabolites of rifabutin, a new antimycobacterial agent, in human volunteers. AB - 1. Metabolites of the antimycobacterial agent 4-deoxo-3,4-[2-spiro-(N-isobutyl-4 piperidyl)]-(1H)-imidazo-(2,5-dihydro )- rifamycin S (rifabutin) were isolated from human urine after administration of a single oral dose of the drug. Some of these metabolites were identified by direct inlet mass spectrometry, 1H-n.m.r. spectrometry and, in two cases, by chromatographic comparison with reference compounds. 2. Unchanged drug, 25-O-deacetyl rifabutin and four other metabolites were identified in human urine. 25-O-Deacetyl rifabutin was the main urinary metabolite, other metabolites were characterized as oxidized, and oxidized deacetylated derivatives. 3. Routes of metabolic transformation were: (a) deacetylation at position 25, (b) oxidation of methyl groups 31 or 32 or at the piperidine nitrogen, and (c) combination of these. PMID- 2549735 TI - Differential diagnosis of human retrovirus infections in the laboratory. AB - Several human retroviruses have been discovered in the past decade, the majority of which have been implicated as etiological agents of severe disease entities. Thus, there is a clear need for accurate identification of human retroviruses in the laboratory. In this review, the classification and general properties of human retroviruses are outlined. Methods for detecting the presence of antibodies are reviewed. In addition, the principles of methods used for isolating and identifying retroviruses are discussed. Finally, techniques which detect the presence of retroviruses directly in clinical specimens without prior amplification in culture are summarized. Clearly, the ability to differentiate and recognize the different types of retrovirus is important for the proper treatment of diseases caused by these viruses. PMID- 2549736 TI - Sexually transmitted viruses. AB - Human viruses known to be spread by sexual contact include herpes simplex viruses (HSV), papillomaviruses (HPV), human immunodeficiency virus (HIV), hepatitis B virus, and cytomegalovirus. Infections with the first three (HSV, HPV, and HIV) have reached epidemic proportions and pose global health concerns. Most of what we know about these human pathogens has been learned only recently, owing to the advent of DNA technologies and advances in culture techniques. In fact, our awareness of one virally transmitted venereal disease, acquired immunodeficiency syndrome, dates to the early 1980s. This paper touches on various aspects of the biology, pathogenesis, clinical manifestations, and, where applicable, oncogenicity of these agents, as well as current treatments and vaccine initiatives. PMID- 2549737 TI - Cytomegalovirus encephalitis: neuropathological comparison of the guinea pig model with the opportunistic infection in AIDS. AB - The AIDS epidemic has transformed the importance of cytomegalovirus (CMV) as a pathogen for the adult human central nervous system (CNS). At autopsy, about 25 percent of AIDS cases have cytopathologic evidence of CNS infection by CMV. Since almost nothing is known of the host CNS-viral interactions, we have developed a laboratory model of CMV infection of the brain in the guinea pig. In the present paper, we review the syndromes of CMV infection of the human CNS and compare the neuropathological findings of the opportunistic CMV brain infection in AIDS with the model. Destructive meningoencephalitis, perivascular infiltrates, and subependymal inflammation are found in both, but the glial nodule is the most characteristic feature of each. Thus, we demonstrate that the model faithfully reflects the histopathology of the human disease. Furthermore, since we have found that CNS infection is achieved following systemic infection in the guinea pig, the model recapitulates the sequence of infection in humans. PMID- 2549739 TI - [Gangliocytic paraganglioma of the duodenum]. AB - In a 49-year-old patient who was investigated because of abdominal complaints we found a polypoid tumor in duodenum near the papilla of Vater. The histological investigation after endoscopical polypectomy showed the rare case of a gangliocytic paraganglioma. After complete endoscopical removal no further intervention is necessary. PMID- 2549738 TI - The switch between EBV latency and replication. AB - This paper reviews experiments done in the author's laboratory which led to the discovery of an Epstein-Barr virus (EBV) gene, BZLF1, whose product, ZEBRA, switches the virus from latency to the replicative phase of its life cycle. Recent experiments are summarized which explore the effects of EBV genome rearrangements and cell background on the expression of ZEBRA, and which investigate the viral targets of ZEBRA action. PMID- 2549740 TI - [Fibrolamellar liver cell cancer. A case report]. AB - Fibrolamellar hepatocellular carcinoma of the liver is a rare variant of hepatocellular carcinoma with characteristic morphological patterns and a good prognosis. Preoperatively the tumor is rarely diagnosed. Surgical treatment is resection, hemihepatectomy or transplantation of the liver. We report a case of a 51 years old patient with fibrolamellar hepatocellular carcinoma of the liver. PMID- 2549741 TI - [Alpha-MSH and pregnancy]. AB - In 32 pregnant women, we repeatedly determined the levels of alpha-MSH, melatonin, ACTH and progesterone. In the radioimmunoassay, all of these hormones showed a significant increase from the first to the third trimester. In contrast to progesterone, neither the level of alpha-MSH nor that of melatonin rises significantly until between the 28th and 36th week of pregnancy. The authors suggest that the increasing pigmentation may be due to the rising levels of alpha MSH, while the preferential hyperpigmentation of sexual areas in additionally mediated by sex hormones. PMID- 2549742 TI - Morphofunctional research on the effects of steroid-stimulating and -inhibiting drugs on the major salivary glands of rats. AB - Adult male albino rats underwent chronic treatment with synthetic adrenocorticotropin and Dexamethasone, and were examined for a long time after the administration of these drugs. The major salivary glands were observed by means of histological and histochemical methods. We did not find enzymatic modifications whereas morphological damage was particularly evident in the submaxillary and parotid glands, especially all after Dexamethasone treatment; this damage is to be found up to at least 14 days after the end of treatment. PMID- 2549743 TI - [The humoral immune response to Epstein-Barr virus in rheumatoid arthritis]. AB - Antibodies to EB virus capsid antigen were assayed for by means of indirect immunofluorescence technique in the serum of 72 rheumatoid arthritis patients; the synovial fluid was tested in parallel with the serum in 18 of these patients. An authentic increase in antibody levels was revealed, as compared to healthy individuals, which is evidence of latent infection activation by EB virus. There was no correlation between the degree to which this activation was pronounced, the rheumatoid factor level in the serum and synovial fluid, and the signs of systemic disease. Half of the rheumatoid synovitis patients were found to have an authentic prevalence of antiviral antibody level in the synovial fluid over that in the serum. PMID- 2549744 TI - Alteration of intracellular monovalent cation concentrations by a poliovirus mutant which encodes a defective 2A protease. AB - Poliovirus mutant 2A-1, which encodes a defective protease 2A, fails to inhibit translation of capped mRNAs selectively. Despite the failure of 2A-1 to inactivate cap-dependent translation, a reduction in the overall rate of protein synthesis, both virus and cell-specified, does occur after 2A-1 infection. This global reduction in protein synthesis is temporally correlated with an increase in [Na+]i and a decrease in [K+]i. The extensive global shutoff of protein synthesis is not observed in 2A-1 infected cells incubated in low NaCl medium or medium containing an elevated concentration of KCl which compensate for the virally-induced alterations in intracellular monovalent cation concentrations. Furthermore, 2A-1-specified protein synthesis is only partly resistant to hypertonic NaCl media which increase [Na+]i, in contrast to protein synthesis specified by wild-type poliovirus. These results suggest that shutoff of host and viral protein synthesis during infection by poliovirus mutant 2A-1 is a consequence of the virus-induced changes in intracellular monovalent cation concentrations. PMID- 2549745 TI - Nucleotide sequence of coronavirus TGEV genomic RNA: evidence for 3 mRNA species between the peplomer and matrix protein genes. AB - The region of the TGEV genome between the E1-matrix protein gene and the E2 peplomer protein gene has been sequenced from a cDNA clone. The consensus recognition sequence, 5'TTAA CTAAAC was found upstream from 3 large open reading frames. In coronaviruses these homologous recognition sequences are involved in the initiation of transcription suggesting that there are 3 mRNA species in this region of the TGEV genome. Northern blot analysis and nuclease S1 mapping confirmed the presence of 3 mRNA species between mRNA 3 encoding the E2-peplomer protein and mRNA 6 encoding the E1-matrix protein. The 5' regions of these 3 mRNAs encode potential polypeptides of predicted molecular weight; 7859, 27744 and 9287, respectively. The potential translation product of ORF B (27744 Da) is considerably larger than previously reported and could be difficult to distinguish by size from the E1-matrix protein. PMID- 2549746 TI - Bluetongue virus genetics and genome structure. PMID- 2549747 TI - Altered hepatitis A VP1 protein resulting from cell culture propagation of virus. AB - The published sequence of hepatitis A virus (HAV), strain HAS-15, after 20-30 cell culture passages contains an 18 nucleotide deletion (Ovchinnikov et al., 1985) within the VP1 genome region. This results in a significant amino acid difference of the VP1 protein when this strain of HAV is compared with other published HAV sequences. Comparison of the polyacrylamide gel electrophoretic migration of HAS-15 HAV and two other strains of HAV revealed that the HAS-15 VP1 molecule migrated faster than the VP1 molecule of the other two strains. Enzymatic amplification of viral RNA derived from the original stool suspension and cell culture adapted HAS-15 using the polymerase chain reaction followed by hybridization analyses with selected synthetic oligonucleotide probes revealed that the original wild type virus did not contain the deletion. These results confirm that cell culture adapted HAS-15 contains an eighteen nucleotide deletion which apparently was selected during cell culture adaptation. PMID- 2549748 TI - Neural infection in mice after cutaneous inoculation with HSV-1 is under complex host genetic control. AB - The ability to restrict neural infection with herpes simplex virus is of potential importance because the nervous system is the main reservoir of virus between recrudescences. Genetic control of innate resistance to herpes simplex after intraperitoneal challenge is well established, but this route of infection does not mimic the progress of virus from skin to the sensory nervous system which occurs during natural infection. We show here, by Mendelian analysis, that the control of neural infection after cutaneous inoculation is complex, involving several (perhaps four) genetic loci, and is not accurately represented by genes which determine survival. The results are discussed in relation to possible underlying resistance mechanisms, particularly the control of early interferon production. PMID- 2549749 TI - [Introduction of an enzyme method for determining pyrophosphate in urine--a comparative study between patients with calculi and normal probands]. AB - Interest aroused towards pyrophosphate as a possible inhibitor in calcium oxalate and phosphate lithiasis. A procedure for the quantitative determination of pyrophosphate in urine suitable for use in clinical laboratories is described. The reproducibility of the method was assessed by within-run and between-run reproducibility studies and the accuracy of the method was obtained by determining the amount of pyrophosphate recovered in urine samples. The present study compares the pyrophosphate excretion and concentration between a control group and a calcium oxalate stone former group. PMID- 2549750 TI - [The morphology of coxsackie B virus infections of the nervous system]. AB - Neuropathological studies were conducted into 20 virologically secured Coxsackie B virus infections of infants and adults, with the nervous system being involved. Inflammatory processes affect both gray and white cerebral regions as well as the peripheral nervous system, and in adulthood they may take the form of severe necrotising encephalitis. Necroses of that kind may be recordable from various regions of the central nervous system. Notwithstanding tempestuous progress in the development of monoclonal antibody kits, culturing of Coxsackie viruses will continue to be of substantive importance to diagnosis, because of the small size of pathogens. PMID- 2549751 TI - [Phylogenetic characteristics of the structure and function of polypeptide growth factors localized in the blood platelets of mammals]. PMID- 2549753 TI - In vitro effects of ketoconazole on corticotrope cell morphology and ACTH secretion of two pituitary adenomas removed from patients with Nelson's syndrome. AB - The direct effects of ketoconazole on the secretion of ACTH by human pituitary adenoma cells from 2 patient with Nelson's syndrome were studied in vitro. Stereologically quantified, intracellular changes affect the surface density of the endoplasmic reticulum (it decreased by 73%), the volume density of the secretion granules (it decreased by 49%), and the volume density of lysosomes (it decreased by 67%). The hormone released in the culture medium decreased depending on the doses of ketoconazole used; 10 mumol/l decreased ACTH levels by 31%. These data show that ketoconazole induce marked changes on corticotrope morphology and ACTH secretion in pituitary cells obtained from patients with Nelson's syndrome. PMID- 2549752 TI - [The biochemical adaptation of the photoreceptor membrane proteins in vertebrate retinal rods to temperature conditions as the evolutionary mechanism maintaining homeostasis at the cellular level]. PMID- 2549754 TI - Stimulatory and inhibitory influences of serum from pregnant women on aromatase activity of immature rat Sertoli cells. AB - Effects of serum from pregnant women on basal and FSH (or cAMP) stimulated aromatase activity of immature rat Sertoli cells in primary culture were studied. Pregnancy serum caused a dose-dependent stimulation of Sertoli cell aromatase activity and the response curves were parallel to those obtained with human FSH. This stimulatory (FSH-like) activity increased progressively during pregnancy, with a sharp drop immediately after delivery. However, the FSH-like bioactivity was not associated with immunoreactive FSH when a specific radioimmunoassay was employed. On the other hand, serum from pregnant women caused a dose-dependent inhibition of FSH and dibutyryl-cAMP-stimulated aromatase activity. These data suggest that human pregnancy serum contains factor(s) which may stimulate basal aromatase activity of Sertoli cells and may inhibit FSH-induced aromatase activity. These factors, most probably of placental origin, may play a role in the regulation of estrogen production during gestation. PMID- 2549755 TI - Heterogeneity of pituitary adenoma cell subpopulations from acromegalic patients obtained by Percoll density gradient centrifugation. AB - Pituitary adenoma cells from 6 acromegalic patients were separated on continuous Percoll density gradients according to differences in their density. Two adenomas produced GH only in culture, the other 4 adenomas produced either GH and PRL (one adenoma) or GH and alpha-subunit (one adenoma) or GH, PRL and alpha-subunit (2 adenomas). The cell subpopulations obtained by this technique differed in the amount of hormone production per 10(5) cells: GH release decreased from the low density fractions to the higher density fractions in 5 of 6 adenomas. Intracellular GH levels completely followed this profile. In the mixed GH/alpha subunit adenomas the alpha-subunit profile completely paralleled the GH profile, whereas in the mixed GH/PRL adenomas the PRL profile showed a pattern different from that of GH (and alpha-subunit). In neither of the adenomas did we find any differences between the subpopulations with respect to the responsiveness of GH, PRL or alpha-subunit release to GHRH, TRH and the somatostatin analogue SMS 201 995. CONCLUSIONS: 1. Within pituitary adenomas from acromegalic patients heterogeneity exists with respect to hormone production per cell. 2. The cell subpopulations obtained by density gradient centrifugation are not different in their responsiveness to SMS 201-995, GHRH or TRH. 3. Because GH and alpha-subunit release by the fractions from the mixed GH/alpha-subunit secreting adenomas were completely parallel, further evidence for co-release of GH and alpha-subunit by the same tumoural cells is provided. PMID- 2549756 TI - Inhibition of insulin degradation by insulin-like growth factors I and II in human hepatoma (HepG2) cells. AB - IGF-I infused at pharmacological doses in healthy men markedly decreases C peptide levels, whereas insulin levels remain within the normal range. One possible explanation is decreased insulin removal. As the liver is the major site of insulin degradation, we studied insulin degradation by HepG2 cells in the presence of IGF. We found that IGF-I at a concentration of 130 nmol/l inhibits insulin degradation by HepG2 cells when the initial insulin concentration is 0.34 nmol/l. The effect of IGF-I on insulin degradation is dose-dependent and the rate of insulin degradation is dependent on the insulin concentration. IGF-II is 6 to 10 times more potent than IGF-I in inhibiting 125I-insulin binding to HepG2 cells and in protecting insulin from being degraded. Thus, IGF-I and IGF-II inhibit insulin degradation most likely by competing for binding at insulin binding sites of liver cells. PMID- 2549757 TI - Aplastic crisis due to human B19 parvovirus infection in red cell pyrimidine-5' nucleotidase deficiency. AB - Two siblings with chronic hemolytic anemia due to red cell pyrimidine-5' nucleotidase (P-5'-N) deficiency, presented within a few days of each other with a febrile illness and pancytopenia. The cause of the aplastic crisis was an acute infection with human B19 parvovirus (B19 HPV) as proven by immunoelectron microscopy and DNA hybridization. This is the first report on the association of B19-HPV-related aplastic crisis with P-5'-N deficiency. PMID- 2549758 TI - Electron microscopical study of non-specific cholinesterase activity in simple lamellar corpuscles of glabrous skin from cat rhinarium: a histochemical evidence for the presence of collagenase-sensitive molecular forms and their secretion. AB - The distribution of nCHE activity was studied histochemically in simple lamellar corpuscles (SLCs) of glabrous skin from cat rhinarium. The Schwann cells forming myelin sheaths in preterminal part of SCLs exhibited no positive reaction for nCHE activity. Prevalent reaction product was localized extracellularly in the inne core enveloping terminal portion of unmyelinated sensory axon. A dot-like shaped reaction product was deposited in the basal lamina of the inner core cells and their cytoplasmic lamellae or was scattered in enlarged interlamellar spaces. Only small amount of fine end product was found to be associated with the plasma membrane of inner core lamellae. Fine reaction product for nCHE activity was consistently localized in perinuclear and rER cisternae and saccules of the Golgi apparatus of inner core cells. Some vesicles around rER and the Golgi apparatus, ones beneath the plasma membrane, and tubular-like cisternal profiles oriented towards the surface contained nCHE end product, as well. The intracellular and extracellular localization of nCHE reaction product suggests that this enzyme behaves in cat SLCs as a secreted rather than as an integral membrane protein. A large amount of dot-like reaction product in the interlamellar spaces disappeared if the skin sections were treated with collagenase before incubation in the medium for histochemical detection of nCHE activity. The decrease of nCHE end product in SLCs of the skin sections after collagenase digestion was corroborated by means of light microdensitometer and electrometrical measurement. The histochemical detection and electrometrical measurement revealed that the majority of the reaction product in the interlamellar spaces of inner core corresponds with the nCHE molecules sensitive to collagenase treatment and they are probably counted among asymmetrical molecular forms. PMID- 2549759 TI - Endothelial pump function in rabbit corneas kept under eye bank conditions. AB - Endothelial pump activity of rabbit corneas kept under eye bank conditions was examined. Different swelling rates for medium with and without ouabain were used for the calculation of endothelial pump rate. After an initial weight increase during 24 h, the control corneas kept approximately constant weight for 1 week. The ouabain-treated corneas increased continually, and during the time interval 24-48 h the increase was 41% of start weight. This corresponds to a pump activity of 2.3% of the weight of the normal corneal stroma per h, or to 9 microns of the corneal thickness per h. Each endothelial cell pumped fluid corresponding to 200 300% of the cell volume per h between day 1 (24 h) and day 8. The pump activity was gradually reduced after 8 days. The average weight was the same in both groups after 7 weeks. PMID- 2549760 TI - Ocular complications after intracarotid BCNU for intracranial tumors. AB - 112 patients with intracranial tumors treated with intracarotid BCNU to a cumulative dose of 370 mg/m2 body surface have been examined retrospectively for ocular complications. Average follow-up was 494 days (range 5 months-5 years). The rate of ocular complications was 2.7%. There was 1 case of hemorrhagic glaucoma with amaurosis and an accompanying oculomotor palsy, 1 case of retinal branch artery occlusion combined with third nerve paresis and 1 case of oculomotor palsy. PMID- 2549761 TI - Determination of Arduan and its desacetyl metabolites in biological fluids. AB - The authors elaborated a method for the determination of Arduan and its desacetyl metabolites in biological fluids. The method is based on the use of labelled Arduan and on the determination of radioactivity of the parent drug and metabolites separated by ion-pair TLC in the development system of chloroform dichloromethane-methanol 6 : 2 : 2 (v/v) 3% NaI (w/v). PMID- 2549762 TI - The effect of CNS depressants on GABA receptor chloride ionophore complexes. AB - Specific binding of 35S-TBPS was examined in synaptosomal membranes of rat cerebral cortex. Various CNS depressants enhanced allosterically both on and off rates of TBPS binding to the convulsant sites of the GABAA receptor chloride ionophore complex. Presumably they facilitate the opening of the chloride channels. PMID- 2549763 TI - Significance of the cerebral dopaminergic neurotransmission for the individual differences in learning and memory (experiments on rats). AB - Using the step-down method for passive avoidance with punishment electroshock reinforcement, the experimental rats were divided according to their individual differences in the capacity for learning and retention into two groups: good learners (GL) and poor learners (PL). Stereotypy and catalepsy methods were used in order to clarify the role of the central dopaminergic (DA-ergic) transmitter mechanisms for the individual differences. Stereotypic motor behaviour was induced by applying the following DA-ergic agonists: apomorphine (2 mg/kg. i.p.), amphetamine (6 mg/kg, s.c.) and elymoklavine (2 mg/kg, i.p.). The stereotypic behaviour was recorded on a 5-point scale. A set of catalepsy tests was used to determine the cataleptic effect of the DA-ergic antagonist haloperidol (1 mg/kg, i.p.). Our experimental results showed that the intensity and duration of the stereotypy caused by the three agents were significantly increased in GL compared with PL. Catalepsy in the PL group appeared later than in the GL group, and its total duration was shorter. The data obtained suggest that in the GL group there is a marked tendency towards a higher level of sensitivity of the brain dopamine receptors, compared with the PL group. PMID- 2549764 TI - Changes in transport functions of isolated rabbit choroid plexus under the influence of oestrogen and progesterone. AB - Isolated choroid plexuses from rabbits were used to determine uptake and accumulation of 10(-5) M radiolabelled choline (expressed as tissue/medium ratio) and the activities of various types of ATPases (based on ouabain inhibition and bicarbonate stimulation) following pre-treatment of the animals with 0.5 mg kg-1 17-beta-oestradiol, alone or in combination with 2 mg kg-1 progesterone. The combined treatment reduced the choline uptake by 35% and also lowered the activity of Na+,K+-ATPase by 31%, without influencing tissue wet weight. The reduction in HCO3-ATPase was smaller and not statistically significant. There was a tendency also for oestrogen alone to lower these activities, but only by less than 20%. The Ca2+,Mg2+-ATPase activity was not significantly affected by any of the hormones. PMID- 2549765 TI - [Paraquat and diquat: mechanisms of toxicity]. AB - The authors review the mechanisms of paraquat and diquat toxicity. They discuss the generation of multiple toxic active forms of oxygen. The pulmonary concentration of paraquat seems to be due to a mechanism of active concentration, which seems to be used by polyamines and diamines in normal situations. Considerations on therapeutics attempts are made. PMID- 2549766 TI - Incidence of late radiation necrosis with transient mass effect after interstitial low dose rate radiotherapy for cerebral gliomas. AB - Late radiation necroses constitute a hazard in low dose rate interstitial irradiation for inoperable gliomas. An incidence of 40% (8/20 patients) was found after permanent implantation of Iodine-125 seeds. This finding may even underestimate the real frequency, because follow-up of unaffected patients was shorter than in patients with radiation necrosis. The necrotic reactions caused a transient mass effect, which lead to a significant deterioration of performance scores. Further manifestations of late delayed radiation damage were observed in two patients. The occurrence of radiation necrosis was correlated with total radiation dose, amount of implanted radioactivity, and with velocity of tumour shrinkage. A mechanism underlying the development of radiation necrosis is proposed: A rapid shrinkage of tumour after interstitial Iodine-125 implantation may cause a significant irradiation of surrounding brain tissue, which was initially lying outside the target volume. Since most patients affected by radiation necrosis were children or adolescents, the risk of radiation damage should be minimized. This could probably be achieved either by reduction of irradiation dose, or by using temporary implants of Iodine-125. PMID- 2549767 TI - Complete remission of recurrent glioblastoma multiforme following local infusions of lymphokine activated killer cells. Case report. AB - We report the case of a 26-year-old man in whom glioblastoma multiforme had recurred six months following a subtotal resection. Despite radiotherapy and a course of interferon beta and ACNU, the tumour increased in size (to 3 cm) and there was neurological deterioration. Treatment was then initiated with LAK cells, together with ACNU and interferon beta. After three courses of LAK cells, tumour size was markedly reduced, and at about six months the tumour had nearly disappeared on computed tomographic (CT) scans. At one year, and after nine courses of LAK cell therapy (total dose of 2.7 x 10(9) cells) infused via an Ommaya reservoir and supplemented by ACNU and interferon beta, the tumour has disappeared and the patient is considered to be in complete remission since 6 months. This marked response is thought to be due chiefly to LAK cell therapy. The relatively low dose administered was well-tolerated. PMID- 2549768 TI - Intracerebral malignant fibrous histiocytoma at site of a previously excised low grade glioma. AB - The case of a 42-year-old patient is reported who developed an intracerebral malignant fibrous histiocytoma at the site of an oligoastrocytic mixed glioma which had been excised 2 1/2 years previously. Reasons for the extreme rarity of intracranial malignant fibrous histiocytomas, the probability of a traumatic aetiology of this particular tumour, and the possible significance of intratumoural cells positive for glial fibrillary acidic protein (GFAP) are discussed. PMID- 2549769 TI - Tumour recurrence vs radionecrosis: an indication for multitrajectory serial stereotactic biopsies. AB - External RT has been proved to be an important adjuvant to surgery in the treatment of malignant glioma. It has also been demonstrated, that its effect on survival is dose-dependent, although accompanied by a higher morbidity. Intents to localize the field of high dose RT to the tumour area have been performed with the aim to spare damage of the normal brain tissue. Between August 1983 to December 1987, 40 patients with malignant astrocytoma (16 GM, 24 AA) underwent high dose localized hyperfractionated external RT after surgical resection. Patients received 57.6 Gy to the tumour and oedema area associated with a boost localized to the tumour of 7.4, 14.4 or 24 Gy. In the follow-up, 16 patients died with evidence of increase in size of lesion diagnosed by CT/MRI. Since July 1987, 12 patients with recurrence or increase on size of CT/MRI lesion have undergone multitrajectory serial stereotactic biopsies. From the biopsies 8 patients were histologically diagnosed was compatible with radionecrosis. From the 4 recurrences, 2 patients were treated with 125I implants and 1 with new resection. Patients with radionecrosis were treated with corticoides and diuretics, obtaining partial or complete remission of symptoms and decrease in size of CT lesion. Undoubtly, Multiplanar/3D multitrajectory serial stereotactic biopsies play a major role in the follow-up of these patients, and accurate diagnosis need to be established for further treatment therapy. The question remains if these localized boost should be replaced by 3D Multiplanar stereotactic interstitial radiotherapy boost after surgery and conventional radiotherapy. PMID- 2549770 TI - Gd-DTPA-enhanced MR detection of diffuse leptomeningeal metastases. PMID- 2549771 TI - Wallerian degeneration and inflammation in rat peripheral nerve detected by in vivo MR imaging. AB - To investigate the role of MR imaging in wallerian degeneration, a series of animal models of increasingly complex peripheral nerve injury were studied by in vivo MR. Proximal tibial nerves in brown Norway rats were either crushed, transected (neurotomy), or transected and grafted with Lewis rat (allograft) or brown Norway (isograft) donor nerves. The nerves distal to the site of injury were imaged at intervals of 0-54 days after surgery. Subsequent histologic analysis was obtained and correlated with MR findings. Crush injury, neurotomy, and nerve grafting all resulted in high signal intensity along the course of the nerve observed on long TR/TE sequences, corresponding to edema and myelin breakdown from wallerian degeneration. The abnormal signal intensity resolved by 30 days after crush injury and by 45-54 days after neurotomy, when the active changes of wallerian degeneration had subsided. These changes were not seen in sham-operated rats. Our findings suggest that MR is capable of identifying traumatic neuropathy in a peripheral nerve undergoing active wallerian degeneration. The severity of injury may be reflected by the corresponding duration of signal abnormality. With the present methods, MR did not distinguish inflammatory from simple posttraumatic neuropathy. PMID- 2549772 TI - Superselective arterial BCNU infusion in the treatment of patients with malignant gliomas. AB - Retinal and CNS toxicity have been reported with infraophthalmic infusion of BCNU in the treatment of patients with malignant gliomas. It is known, however, that the CNS toxicity can be reduced if the BCNU is dissolved in dextrose in water. This article describes the results from 15 patients who received 42 courses of BCNU administered by supraophthalmic internal carotid, middle cerebral, or posterior cerebral artery infusions. None of the patients developed leukoencephalopathy as demonstrated by CT scanning. The average reduction in tumor volume was 36%, and the median survival time from the date of diagnosis was 73 weeks. These values are comparable to those of a previous group of 20 patients treated with infraophthalmic infusions, with the exception that none of the patients in the present group developed retinal damage. PMID- 2549774 TI - Ganciclovir approval. PMID- 2549773 TI - Beta-adrenergic receptors in nasal tissue with reference to their role in asthma and atopy. AB - Thirty-three samples of nasal tissue (turbinates and polyps) were collected and assayed for beta-adrenergic receptor (BAR) sites (radio-ligand assay). Patients were divided into groups: 1) nasal turbinates, 2) nasal polyps, both under the aspects of atopy and bronchial hyperreactivity. Different radiobinding was found in both groups: Patients A with high BAR and patients B with low BAR numbers. There was no correlation between BAR numbers--clinical symptoms and bronchial hyperreactivity. Generally the BAR numbers were higher in turbinates (p less than 0.01) than in polyps and in nonatopic compared to atopic patients (p less than 0.005). "Affinity state 1" BAR are equal in all groups; however, "affinity state 2"BAR are significantly less in polyps (p less than 0.05). In 50% of the examined cases there was a combination of polyps, asthma and atopy putting foreward a common line of pathogenesis in this entity. It is discussed that the underlying inflammation (allergic and non-allergic) is the cause for reduced BAR numbers in the effected tissue. So far nasal tissue seems to be like a mirror compared to the results from lower respiratory tract tissue with respect to beta-adrenergic receptors. PMID- 2549775 TI - Depressant effect of magnesium on early afterdepolarizations and triggered activity induced by cesium, quinidine, and 4-aminopyridine in canine cardiac Purkinje fibers. AB - Magnesium chloride has been shown to terminate torsades de pointes in some patients with the acquired long QT syndrome. The mechanism for this effect is unknown. Recently early afterdepolarizations (EADs) and triggered activity (TA) have been proposed as causes of torsades de pointes. The purpose of the present study was to examine whether magnesium suppressed EADs that were initiated in vitro by different agents and if so its mechanism of action. TA arising from EADs was induced by quinidine (1 to 4 mumol/L, n = 5) at high temperature (38.5 to 40 degrees C), cesium chloride (5 to 12 mmol/L, n = 6), and 4-aminopyridine (1.5 to 5 mmol/L, n = 7) in canine cardiac Purkinje fibers superfused with modified Tyrode's solution (KCI = 2.7 mmol/L). MgCl2 (2 to 7 mmol/L) reversibly abolished TA and suppressed EADs. Tetrodotoxin (TTX; 1 to 5 mumol/L) also abolished TA elicited by 4-aminopyridine (n = 6). We then examined the effects of MgCl2, TTX, and verapamil on depolarization-induced automaticity by means of a single sucrose gap technique to gain insight into the mechanism of action of magnesium. MgCl2 (5 mmol/L) abolished automaticity arising from membrane potentials more negative than -70 mV and prolonged the spontaneous cycle length at less negative membrane potentials. The effects of TTX (1 to 5 mumol/L) resembled those of MgCl2. Verapamil (1 mumol/L) prolonged the cycle length of the initial automatic response at high levels of membrane potential and progressively reduced the amplitude of the subsequent automatic potentials. It abolished automaticity arising from less negative membrane potentials.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549776 TI - Localization and quantification of hormones, ligands, and mRNA with affinity-gold probes. AB - The application of immunogold techniques to localize pituitary hormones produces label that can be quantified and correlated with different secretory states. This report focuses on three major applications of the technology. In the first set of studies, immunogold labels for adrenocorticotropin (ACTH) or luteinizing hormone (LH beta) and follicle-stimulating hormone (FSH beta) were applied to ultrathin sections of pituitaries from adrenalectomized rats or from rats in different stages of the estrous cycle. During the first week after adrenalectomy, ACTH cell area increased. The concentration of immunoperoxidase label (amount of label/area of the corticotropes) decreased. Counts of gold markers showed that there were no changes in the concentration of antigens per granule. Three weeks after adrenalectomy, the amount of immunoperoxidase label increased along with the concentration of that label. The concentration of gold label for ACTH on granules also increased. All changes correlated well with increases in serum ACTH stimulated by adrenalectomy. In the studies of cycling rats, gonadotropes showed increases in the number of gold markers for LH beta or FSH beta per granule area just before an elevation in serum levels. There were also increases in the proportion of granules that contained only LH beta or FSH beta (monohormonal) before the rise in secretion. Thus, nonparallel release of gonadotropins might be attributed to changes in the ratio of gonadotropins packaged per granule. In the second series of studies, avidin-gold labels were used to identify sites of binding of biotinylated ligands. These studies illustrate and quantify binding by biotinylated gonadotropin-releasing hormone (GnRH) to ovarian or pituitary target cells. Triple-labeling protocols (avidin-peroxidase followed by immunogold) show that the target cells in the pituitary contain gonadotropins. In the third set of studies, avidin gold or avidin peroxidase was used to label sites of hybridization of a biotinylated cRNA probe to gonadotropin beta subunit mRNA. The sites of hybridization appear on rough endoplasmic reticulum; however, further work is needed to improve cell ultrastructure and perserve antigens. Triple labeling protocols (avidin-peroxidase followed by immunogold) show the feasibility of the technique as well as the need for further refinement. To summarize, these studies describe multiple applications of gold labels for the localization of antigens, ligands, and mRNA. The labels are sensitive for detection of antigens and ligands and easily quantified. Quantitative analyses show changes in concentration of gold label that correlate well with secretory states. PMID- 2549777 TI - Applications of immunogold-silver enhancement: testing of monoclonal antibodies and detection of human cytomegalovirus in histologic specimens. AB - Human cytomegalovirus (HCMV) is an important pathogen in neonates, transplant patients, and individuals with acquired immunodeficiency syndrome (AIDS). Reliable techniques for the detection of this virus in clinical specimens would aid in improving methods for diagnosis and increasing our understanding of viral pathogenesis. We evaluated the utility of immunogold-silver enhancement to determine the specificity of murine monoclonal antibodies directed against HCMV proteins and applied these antibodies to the detection of HCMV in histologic material. Nine antibodies were tested in a tissue-culture system to determine the location of staining. All were found to be active in frozen tissue, but only two of these antibodies were reactive in formalin-fixed tissue. We also evaluated a novel tissue fixative technique (AMeX; Fixation and dehydration with acetone) that has been used to maintain antigenicity of T-lymphocyte cell markers. All antibodies remained reactive against their respective HCMV proteins in tissue fixed by this technique. However, dehydration of tissue may limit the usefulness of AMeX fixation. Immunogold-silver enhancement is a useful and reliable histochemical technique for detection of HCMV antigens in pathologic specimens. PMID- 2549778 TI - Use of immunogold preembedding technique to detect hepatitis A viral antigen in infected cells. AB - Localization of virus and viral antigen in cell cultures infected with a rapidly replicating isolate of strain HM-175 of hepatitis A virus (HAV; pHM-175) was accomplished by using immunogold probes. Cells infected under one-step growth curve conditions were prefixed with 2% paraformaldehyde and 0.1-0.001% saponin at appropriate times postinfection for detection of maximum virus and viral antigen. An indirect labeling technique was employed using monoclonal antibody to HAV followed by 5 nm gold-antimouse IgG conjugate. Cells were then fixed by standard electron microscopy techniques and thin sectioned. This prefixation technique allowed penetration of the immunogold probes and moderate preservation of ultrastructure. Within infected cell cytoplasm, numerous antigenic sites were labeled with six to 200 gold particles. Two types of cells were infected with HAV and somewhat different results were obtained with the two cell types. In BS-C-1 cells, where a cytopathic effect (CPE) was not observed, myelin figures were immunogold labeled or frequently were located near immunogold-labeled sites. Vesicles containing viruslike particles (14-22 nm) were also observed. A significant observation in infected FRhK-4 cells was the presence of multivesicular bodies labeled with immunogold. Microfilaments were commonly seen near the multivesicular bodies. Our results demonstrate that the choice of prefixation method for immunogold labeling should be empirically determined for the cell type and condition. PMID- 2549779 TI - Recovery of soluble dietary fiber is dependent on the method of analysis. AB - The effects of different methods on the distribution of total neutral sugars (TNS), uronic acids (UA), and beta-glucans (beta G) between the soluble (S) and insoluble (I) fractions of dietary fiber (DF) were determined for peas, kidney beans, oat bran, rice, and macaroni. Incorporation of a protease step into the Theander method "A" modestly increased, and addition of a pepsin digestion further increased the proportion of total fiber recovered in the S fraction. The effect of extraction method on the distribution of TNS, UA, and beta G between the S and I fractions varied with the food. The three methods measured the same total DF in a food and 1-3% starch in the I fraction of peas and kidney beans. Use of dimethyl sulfoxide to solubilize starch, or elevated temperature to extract S components, had no effect on the distribution of DF between S and I fractions of peas and macaroni. Incomplete protein hydrolysis did not always lower Klason lignin and excluding lignin from the fiber complex did not always substantially increase the S fraction. PMID- 2549780 TI - Soy-polysaccharide fiber: effect on diarrhea in tube-fed, head-injured patients. AB - In a randomized, double-blind, crossover study the effect of tube-feeding with soy-polysaccharide fiber (SPF) vs without SPF on stool weight (SW), stool consistency (SC), fecal nitrogen (FN), and incidence of diarrhea (ID) was compared in nine head-injured subjects; associations with feeding volume, albumin level, and medications were also examined. Subjects with and without SPF had (mean +/- SEM) SW of 245 +/- 47 and 277 +/- 49 g/d, SC of 2.17 +/- 0.01 and 2.52 +/- 0.13 (3 is watery) (p less than 0.01), FN of 1.35 +/- 0.45 and 1.36 +/- 0.33 g/d, and ID of 33% and 44%, respectively. Significant treatment X treatment sequence interaction nullified results for SW and SC. Neither FN nor ID were affected by SPF. The condition of all variables tended to improve over time. Certain medications and SPF were predictive of SW and SC; albumin and tube feeding volume were not. SPF-containing tube feedings did not seem to have an effect on bowel function in these well-nourished head-injured patients. PMID- 2549782 TI - Long-term surviving patients with hepatocellular carcinoma comprised of dense collagenous stroma. AB - In two patients with hepatocellular carcinoma (HCC), treated surgically, we observed a dense collagenous stroma not heretofore reported. Grossly, the tumors consisted of a broad area of dense collagenous stroma that embedded the nodules of the HCC. Microscopically, nodules of the cancer cells were widely separated by this dense fibrous stroma, in association with surrounding inflammatory infiltrates, composed mainly of lymphocytes. These two patients had lived, respectively, for 10 yr and 2 months, and 8 yr and 9 months, despite precarious clinical states. HCC with characteristics such as a grossly distinct dense collagenous stroma and an inflammatory infiltrate may relate to a distinct category with a favorable prognosis. PMID- 2549781 TI - Identifying high-risk precursors of cervical cancer. How and why. PMID- 2549783 TI - Hepatocellular carcinoma presenting with the superior mediastinal syndrome. AB - Two southern African black men with hepatocellular carcinoma are described in whom spread of the tumor to mediastinal lymph glands caused obstruction to the superior vena cava. Although necropsy studies have shown that hepatocellular carcinoma occasionally metastasizes to mediastinal lymph glands, presentation with the superior mediastinal syndrome has not hitherto been reported. PMID- 2549784 TI - Hepatocellular carcinoma in a case of familial polyposis coli. PMID- 2549785 TI - Mitomycin C adsorbed on activated carbon particles: is intraperitoneal chemotherapy effective for synchronous pleuritis carcinomatosa? PMID- 2549786 TI - Papillomavirus and esophageal cancer in the Japanese and Chinese. PMID- 2549787 TI - Variation in smoking-related lung cancer risk among New Jersey women. AB - A population-based incident lung cancer case-control study was conducted among New Jersey women in 1982-1983. Interviews were completed for 994 cases and 995 controls. The association of lung cancer with cigarette smoking had an overall age-, race-, and respondent type-adjusted odds ratio of 8.5, with risks of 11.1, 62.6, and 3.9 for squamous cell, small cell, and adenocarcinoma, respectively. Analyses for squamous cell carcinoma showed significantly higher odds ratios for self-respondents than for subjects with next of kin respondents; this heterogeneity may be partially explained by misclassification in next of kin reports. Analyses for adenocarcinoma showed a significantly lower smoking-related risk among older women, which may be related to the later age at which these women began smoking. Analyses by histologic subtype showed lower risks for poorly differentiated squamous cell carcinoma and higher risks for poorly differentiated adenocarcinoma. This could be due to misclassification of histologic type and suggests that the actual smoking-related risk for squamous cell carcinoma might be higher than observed, and that for adenocarcinoma might be lower. Comparison of the results in this study with those from a similar study of lung cancer among men showed little difference between men and women in the magnitude of the association with smoking, or in the patterns of risk with intensity and duration. PMID- 2549788 TI - The Tecumseh Study. XV: Rotavirus infection and pathogenicity. AB - Rotavirus infection and associated symptoms were studied prospectively from 1976 through 1981 in a 10% sample of families in Tecumseh, Michigan. Infection was determined using an antibody enzyme-linked immunosorbent assay (ELISA) method designed for use with large numbers of sera collected semiannually. Illness was identified from symptoms ascertained weekly by telephone. Risks of infection during a rotavirus season varied from 4.8% in 1978 to 9.2% in 1980. Risks were 20.6% in the under two years age group and decreased progressively with age to 3.6% in individuals aged 45 years or over. Preschool children accounted for less than 20% of the infections in the population of reference. Infection was associated with diarrhea in the first two years of life and with both diarrhea and vomiting later in life. There was no evidence for association of rotavirus infection with a gastrointestinal-respiratory syndrome. The pathogenicity of infection, the protectiveness of antibody, and the socioeconomic correlates of infection showed marked changes from year to year. In 1978, there was a dramatically higher pathogenicity and a greater protectiveness of ELISA measured antibody than in other years. There was also a dramatic reversal in the socioeconomic relations of infection from 1977 to 1978. Taken together, these findings suggest that different rotaviruses might have predominated in these years. PMID- 2549789 TI - Recurrent Guillain-Barre syndrome after multiple exposures to cytomegalovirus. PMID- 2549790 TI - Fluconazole treatment of cryptococcosis in patients with acquired immunodeficiency syndrome. PMID- 2549791 TI - Somatostatin analogue: use in the treatment of vipoma with hypercalcemia. PMID- 2549792 TI - Cerebrospinal fluid from cognitively impaired patient with acquired immunodeficiency syndrome shows gp120-like neuronal killing in vitro. PMID- 2549793 TI - Aqueous chamber tap and serology in acute retinal necrosis. PMID- 2549794 TI - Role of xanthine oxidase in thermal injury of skin. AB - These studies were designed to assess pathophysiologic factors responsible for increased vascular permeability occurring in rat skin that has been thermally injured in vivo. Under the conditions employed, permeability changes and edema formation progressed over time, with peak changes occurring 60 minutes after thermal trauma. The plasma of thermally injured rats showed dramatic increases in levels of xanthine oxidase activity, with peak values appearing as early as 15 minutes after thermal trauma. Excision of the burned skin immediately after thermal injury significantly diminished the increase in plasma xanthine oxidase activity. The skin permeability changes were attenuated by treatment of animals with antioxidants (catalase, superoxide dismutase [SOD], dimethyl sulfoxide [DMSO], dimethylthiourea [DMTU]) or an iron chelator (deferoxamine), supporting the role of oxygen radicals in the development of vascular injury as defined by greatly increased vascular permeability. Studies employing laser Doppler velocimetry in thermally injured skin revealed a pronounced and sustained decrease in blood flow after thermal trauma, a pattern not affected by protective interventions. The failure of neutrophil depletion to protect against the vascular permeability changes and the protective effects of the xanthine oxidase inhibitors (allopurinol and lodoxamide tromethamine) suggest that xanthine oxidase is the most likely source of the oxygen radicals involved in edema formation. Lodoxamide was found to have some hydroxyl radical (HO.) scavenging ability (greater than that of allopurinol) but no iron chelating activity. Some of the protective effects of lodoxamide and allopurinol may be linked to their HO. scavenging ability. These data suggest that, in this model of thermal trauma, vascular injury defined by increased vascular permeability is, in part, related to the activation of xanthine oxidase and the generation of toxic oxygen metabolites that damage microvascular endothelial cells. PMID- 2549796 TI - Herpes simplex virus type 2 antibodies: high prevalence in monogamous women in Costa Rica. AB - We studied the prevalence of antibody to Herpes simplex virus types 1 and 2 (HSV 1 and HSV-2) in 766 randomly selected Costa Rican women 25-59 years of age in a national household survey in 1984-1985. Overall, 97.1% were seropositive for HSV 1 and 39.4% for HSV-2. Only 1.1% of HSV-2 seropositive women gave a history of symptomatic genital herpes. HSV-2 virus antibody increased with age and with the number of lifetime sexual partners. HSV-2 seroprevalence among women who reported only 1 lifetime sexual partner was almost twice as high as the prevalence among women who denied sexual experience (30.5% vs. 17.7%) and reached 79.2% among women with greater than or equal to 4 partners. HSV-2 seroprevalence was lower among women whose partners used condoms: 28.9% for those who had used condoms for at least 2 years vs. 44.3% for those who never used condoms. PMID- 2549797 TI - Giant axonal neuropathy. A review. AB - First reported in 1972 by Berg & colleagues, giant axonal neuropathy is a generalized disorder of cytoplasmic intermediate filaments affecting the nervous system particularly. The condition was originally thought to be a disorder of the peripheral nervous system, but clinical and pathological evidence has now accumulated which indicates that the brain and spinal cord are progressively involved. Over 20 cases have been reported to date. All cases reported have developed clumsiness and progressive weakness with hyporeflexia in the first seven years of life. Later dysarthria, cerebellar signs and pyramidal tract disturbances appear. Mental retardation, dementia and seizures are sometimes seen. Tightly curled hair is characteristic of, but not invariably present in, the condition. This disorder, as well as a similar condition affecting dogs, appears to be transmitted by autosomal recessive inheritance. No treatment is effective. Most cases are wheelchair bound or dead by the end of the second decade. PMID- 2549795 TI - Expression of HLA-A, -B, -C, -DR, -DP, -DQ, and of HLA-D-associated invariant chain (Ii) in non-neoplastic mammary epithelium, fibroadenoma, adenoma, and carcinoma of the breast. AB - Non-neoplastic mammary gland, 20 benign tumors and 206 carcinomas of the breast were immunohistochemically examined for expression of HLA-A, -B, -C, HLA-DR, -DP, and -DQ molecules and the HLA-D associated invariant chain (Ii). In contrast to cells from benign lesions, tumor cells of 51.2% of carcinomas had an abnormally low content of HLA-A, -B, and -C determinants ranging from reduction of antigenic density per cell (28.8%) over an incomplete (15.6%) to complete loss of antigens (6.8%). Associated with lymphohistiocytic stromal infiltrates, HLA-D/Ii determinants were found to be induced in benign duct and acinar epithelium after the order Ii greater than or equal to HLA-DR greater than or equal to HLA-DP greater than or equal to HLA-DQ. These antigens were also expressed, mostly noncoordinately, in 55.5% of carcinomas, and in 98 cases according to the above order. In 28.6%, Ii expression clearly exceeded HLA-D antigen expression; conversely, 6.2% contained HLA-DR+/Ii- tumor cell subsets. In breast carcinoma, the association of reduced HLA-A, -B, and -C expression and a noninduction of HLA DR was highly significant (P less than 0.0009), suggesting an abnormal signal acting down-regulating on the expression of both classes of antigens. Because the modality of HLA-A, -B, and -C and HLA-D/Ii expression correlated with neither tumor type nor grade, it might be an independent parameter. PMID- 2549798 TI - Histogenesis of extramammary and mammary Paget cells. An immunohistochemical study. AB - To define the histogenesis of the Paget cell and possibly identify differences in cells from the two sites, six vulvar and 23 mammary specimens from Paget's disease lesions were studied for immunocytochemical antigens. All vulvar and 21 (91%) mammary lesions were strongly reactive for glandular cytokeratin. All lesions showed immunopositive Paget cells with epithelial membrane antigen (EMA). An apocrine antigen, gross cystic disease fluid protein (GCDFP-15), decorated 66.5% and 56.5% of extramammary and mammary lesions, respectively. All vulvar Paget cells stained for carcinoembryonic antigen (CEA), a frequency significantly greater than the 35% in mammary lesions (p = 0.02). However, CEA is expressed by both eccrine and apocrine sweat glands and their tumors. Vulvar Paget cells were negative for lysozyme, casein, lactalbumin, and S100 protein, compared with 9%, 4%, 4%, and 26% in nipple lesions. S100 protein expression is similar to that in mammary ductal carcinoma (32%). The glandular origin of both extramammary and mammary Paget cells is indicated by the presence of glandular cytokeratin, EMA, and CEA. Approximately 60% of all cases in both sites showed evidence of apocrine derivation (GCDFP-15 positivity). Variable antigen expression suggests possible malignant transformation of pluripotent germinative cells able to differentiate in an apocrine or an eccrine direction, or in both. PMID- 2549799 TI - Follicular lymphoid hyperplasia of the skin with high content of Ki-1 positive lymphocytes. AB - We report the case of a spontaneously regressing lymphocytic lesion on the neck of a 76-year-old man. Routine histopathological examination revealed a heterogeneous infiltrate composed of both lymphoid follicles and preferentially perifollicular and interfollicular sheets of large atypical cells. Immunohistologically, large atypical cells were positive for Ki-1/Ber-H2 (CD30), anti-Il-2 (CD25), Ber T9, anti-HLA-DR, Leu-5b (CD2), T3 (CD3), Leu-3 (CD4), Leu-1 (CD5) and, occasionally, Leu-M1 (CD15); that is, they showed an arrangement and immunophenotype similar to those of perifollicular Ki-1 positive cells in reactive lymphoid tissue. In contrast, antibodies To15 (CD22), anti immunoglobulins, and anti-CALLA (CD10) stained polyclonal B-cells restricted to lymphoid follicles and small clusters representing remnants of follicles. We suggest that this lesion be termed "follicular lymphoid hyperplasia of the skin with high content of activated T-helper cells." PMID- 2549800 TI - Malignant dermal cylindroma in a patient with multiple dermal cylindromas, trichoepitheliomas, and bilateral dermal analogue tumors of the parotid gland. AB - A malignant dermal cylindroma of the scalp arose from one of multiple long standing dermal cylindromas in a 76-year-old man with coexisting trichoepitheliomas and bilateral dermal analogue tumors of the parotid gland. The histologic transition from a benign dermal cylindroma to an anaplastic keratinocytic neoplasm was readily apparent. The malignant dermal cylindroma is a rare neoplasm. To our knowledge, the constellation of benign and malignant dermal cylindromas, multiple trichoepitheliomas, and salivary gland neoplasms has not been previously reported. PMID- 2549801 TI - Atracurium infusion during paediatric craniofacial surgery. Closed loop control of neuromuscular block. AB - A simple feedback control system was used to administer atracurium to 10 infants aged between 5 and 36 months during craniofacial reconstruction. The purpose of the study was to establish whether the system, previously evaluated only in adults, would be effective in paediatric patients. The individuals studied underwent long procedures with substantial blood loss and the potential for thermal and electrolyte disturbances. The system maintained the level of neuromuscular block successfully between 88 and 100% in all patients and between 90 and 100% in nine patients. The mean duration of feedback control was 179 (range 111-440) minutes. The mean (SD) dose of atracurium required was 0.40 (0.12) mg/kg/hour. The mean (SD) recovery time to a train-of-four ratio of 0.75 was 24.8 (9.9) minutes. PMID- 2549802 TI - Sodium bicarbonate as a single dose antacid in obstetric anaesthesia. AB - Gastric pH and volumes were measured in 84 women who had general anaesthesia for emergency Caesarean section. Forty-eight received only 20 ml 8.4% sodium bicarbonate immediately before induction of anaesthesia and 36 ranitidine 150 mg 6-hourly during labour in combination with NaHCO3. Gastric pH was less than 2.5 in four women who received only bicarbonate. All aspirates from the ranitidine plus bicarbonate group had a pH greater than 2.5. Mean volumes of gastric content aspirated were 87 (SD 87.4) and 60 (SD 46.3) ml for the bicarbonate alone and bicarbonate plus ranitidine series respectively. These differences were not significant. Twenty millilitres of 8.4% NaHCO3 cannot be recommended as a single dose antacid for emergency Caesarean section. Ranitidine plus bicarbonate is considered a reliable antacid regimen to ensure elevation of gastric pH to safe levels. PMID- 2549803 TI - Anaesthesia for a child with congenital myotonic dystrophy. PMID- 2549804 TI - Preparation of a photoreactive analog of parathyroid hormone [Nle8, Lys(N-epsilon 4-azido-2-nitrophenyl)13,Nle18,Tyr34]bovine parathyroid hormone-(1-34)NH2, a selective, high-affinity ligand for characterization of parathyroid hormone receptors. AB - Photoaffinity radiolabeling techniques have been widely used to characterize the properties of peptide hormone receptors. However, the identity of authentic receptors is often uncertain because many macromolecules are labeled. These ambiguities are due, in part, to the use of a heterogeneous mixture of photoreactive photoligands, many of which have no or low affinity for the relevant hormone receptor. In this report, we describe the synthesis, purification, and structural analysis of the photoreactive parathyroid hormone analog, [Nle8,Lys(N-epsilon-4-azido-2-nitrophenyl)13,Nle18,Tyr34]-bovine parathyroid hormone-(1-34)NH2. The sulfur-free, oxidation-resistant, synthetic analog of bovine parathyroid hormone (PTH), [Nle8,Nle18,Tyr34]bovine PTH-(1 34)NH2 (NlePTH), was reacted with 4-fluoro-3-nitrophenylazide under nonaqueous conditions to yield several derivatives which were separated by reverse-phase high-performance liquid chromatography and analyzed by amino acid compositional analysis, thin-layer chromatography, and ultraviolet and visible absorption spectroscopy. Among the NlePTH derivatives generated, one of the least hydrophobic was shown to retain the highest potency as assessed in the canine renal cortical membrane radioreceptor assay. Sequence analysis of this peptide, after it had been derivatized with 4-fluoro-3-nitro-[2,6-3H]phenylazide and purified to homogeneity, permitted us to determine that the structure of this analog is [Nle8,Lys(N-epsilon-4-azide-2-nitrophenyl)13,Nle18,Tyr34]bovine PTH-(1 34)NH2. We emphasize the importance of using photoreactive ligands which are purified and subjected to detailed chemical and biological analyses for characterizing the properties of parathyroid hormone receptors and receptors for other peptide hormones. PMID- 2549806 TI - Quantitation of leukotriene B4 in human serum by negative ion gas chromatography mass spectrometry. AB - Leukotriene B4 (LTB4) is a potent chemotactic agent formed via the 5-lipoxygenase pathway from arachidonic acid. To understand the role LTB4 plays in several pathological processes it is essential that endogenous concentrations of LTB4 be accurately quantitated. We have developed a method based on electron capture negative ion mass spectrometry for the analysis of LTB4 in serum at low picogram per milliliter concentrations. Blood is collected into the 5-lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA) to suppress ex vivo formation. Serum is isolated, equilibrated with the internal standard [2H4]LTB4, and extracted using octadecyl-silica (C-18) cartridges. After conversion of the carboxylic acids to their pentafluorobenzyl esters the extract is purified by straight-phase HPLC. Gas chromatographic-mass spectrometric analysis is accomplished on the tert butyldimethylsilyl ether derivatives using dual-selected ion monitoring of m/z 431 and 435. These ions correspond to loss of tert-butyldimethylsilanol from the (M-PFB)- ion of endogenous and [2H4]LTB4, respectively. The concentration of LTB4 in human serum samples was 10.0 +/- 4.0 pg/ml (n = 5). The assay exhibited satisfactory precision, with an intraassay coefficient of variation of 17% and a high degree of accuracy. The concentration of LTB4 in serum collected with (NDGA) was less than 10% of that observed in blood collected without the lipoxygenase inhibitor. Ex vivo formation can therefore be a major obstacle in assessing circulating levels of LTB4. PMID- 2549805 TI - Enzymatic synthesis of DNA probes complementary to a human variable number tandem repeat locus. AB - Both cloned and synthetic DNA probes complementary to human variable number tandem repeat (VNTR) loci have been used to detect restriction fragment length polymorphism. In this report, we describe an approach for the enzymatic synthesis of a DNA probe complementary to one VNTR locus. The probe is produced by annealing short synthetic oligonucleotides comprising single repeat units and enzymatically ligating them into a polymeric DNA probe. In HinfI digests of human genomic DNA separated by agarose gel electrophoresis, this ligated oligonucleotide probe (LOP) detects multiple polymorphic loci in the range of 3 23 kb producing highly informative DNA fingerprint patterns when different individuals are compared. The hybridization pattern is very stable even under high-stringency wash conditions. The LOP is more easily generated than cloned VNTR probes and is totally synthetic, avoiding problems associated with cloned probes including bacterial growth and maintenance as well as in vitro labeling. PMID- 2549807 TI - A simple and efficient method for the oligonucleotide-directed mutagenesis using plasmid DNA template and phosphorothioate-modified nucleotide. AB - We have developed a simple and efficient method for oligonucleotide-directed mutagenesis with double-stranded (plasmid) DNA as a template. The template was simply and rapidly prepared by cell lysis and the following DNA denaturation with alkali. The chain elongation was performed with phosphorothioate-modified nucleotide at 37 degrees C. After the selective digestion of original DNA with NciI and exonuclease III, the desired mutated gene was obtained at a high frequency (about 70%). PMID- 2549808 TI - Controlled partial restriction digestions of DNA by competition with modification methyltransferases. AB - Competitive reactions, using defined ratios of DNA restriction methyltransferase to endonuclease, are shown to result in reliable partial restriction digests of DNA. This method is suitable over a wide range of DNA concentrations and works on DNA in liquid or embedded in agarose. Simultaneous methylase/endonuclease reactions using endonucleases that cleave human DNA very infrequently, such as ClaI or NotI, should generate very large discrete partial DNA fragments suitable for physical mapping in the million base-pair range. Another possible application of methylase/endonuclease competitive reactions is the production of defined partial digests for making cosmid, lambda, or other genomic libraries. PMID- 2549809 TI - N-iodoacetyltyramine: preparation and use in 125I labeling by alkylation of sulfhydryl groups. AB - Preparation and use of N-iodoacetyltyramine in generation of 125I-labeled compounds is described. The kinetics of alkylation of N-acetylcysteine by N iodoacetyltyramine (k2 = 3.0 M-1 s-1) and N-chloroacetyltyramine (k2 = 0.12 M-1 s 1) indicate that N-iodoacetyltyramine is more useful for labeling sulfhydryl containing compounds to high specific activity with 125I. Conditions for preparation of carrier-free 125I-labeled N-iodoacetyl-3-monoiodotyramine in 50% yield based on starting iodide are described. The high degree of group specificity of N-iodoacetyl-3-monoiodotyramine reaction with sulfhydryl groups is demonstrated by the high reactivity toward sulfhydryl-containing bovine serum albumin and low reactivity toward N-ethylmaleimide-blocked bovine serum albumin and IgG. 125I-labeled N-iodoacetyl-3-monoiodotyramine was also used to prepare an 125I-labeled ACTH derivative that retains full biological activity, further demonstrating the selectivity toward reactions with sulfhydryl groups. PMID- 2549810 TI - Osteocalcin-hydroxyapatite interaction in the extracellular organic matrix of bone. AB - Osteocalcin, a major noncollagenous matrix protein of bone, dentin, and cementum, is found in tight association with the calcium phosphate mineral phase of these tissues. This article reviews the structural data for osteocalcin relevant to mineral adsorption. The equilibrium-binding properties for Ca2+ ions and hydroxyapatite are considered, along with the apparent physicochemical effects of osteocalcin on bone mineral dynamics. Several of osteocalcin's possible biological activities (involvement in mineralization, chemoattraction, and leukocyte elastase inhibition) are discussed in relation to the mineral adsorption characteristics of this protein. PMID- 2549811 TI - Molecular mechanisms of bone resorption by the osteoclast. AB - The osteoclast is a multinucleated cell that is actively engaged in the synthesis of lysosomal enzymes, their vectorial transport toward the apical membrane, and the secretion of these enzymes at its apical pole. These secreted enzymes are targeted to the apical ruffled-border membrane by mechanisms that involve cation independent mannose-6-phosphate receptors. These receptors bind to an enzyme linked mannose-6-phosphate recognition marker in the Golgi complex, and the enzyme-ligand-receptor complex, carried within small coated transport vesicles, dissociates upon reaching the low pH established in the bone-resorbing compartment by the osteoclast. The apical bone-resorbing compartment is sealed off by the attachment of the osteoclast to the calcified matrix and is actively acidified by the osteoclast. The plasma membrane of the cell is divided into distinct domains. The apical membrane at the ruffled-border shares common antigenic determinants with lysosomal and endosomal membranes, including a 100 kD protein and proton pumps that may be involved in the acidification of the extracellular resorbing compartment. The basolateral membrane is highly enriched in carbonic anhydrase, and bicarbonate-chloride exchange appears to regulate the intracellular pH of this cell. These observations are consistent with a scheme in which, in the low pH environment of the bone-resorbing lacuna produced by the osteoclast, the mineral phase dissolves, exposing the organic matrix to the action of the secreted enzymes. The activity of these enzymes is in turn presumably favored by the acidic milieu. All constituents of the matrix, whether mineral or organic, then would be reduced to their elemental forms (ions and amino acids) extracellularly.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549812 TI - Annulate lamellae-soleplate nuclei associations in skeletal muscle fibers of rats during chronic high-dose exposure to neostigmine. AB - To investigate annulate lamellae (AL) with nuclear changes, ultrastructural time course studies were done of the reversible end-plate myopathy in the soleus muscles of rats exposed chronically to a high dose of the anticholinesterase drug neostigmine. At the earliest stage (2 hours) in which severe subjunctional damage involving a nuclear lesion (nuclear pyknosis) was prominent, AL profiles were undetectable. At the intermediate stage (7, 21 days), in which the subjunctional organelles tended to cluster and nuclear pyknosis often accompanied degenerative features, most AL laid near the surface of the abnormal nuclei, where there were signs of elimination or formation of pores, as well as a progression of changes leading to the loss of pores. At the late stage (56 days), in which muscle repair was nearly completed and euchromatic nuclei usually were found, there was a concomitant enhanced formation of false nuclear inclusions and of the AL profiles in these sites. Pores are packed on nuclear envelopes, which detach themselves from the nucleus and take the same profile as AL. This phenomenon can be envisioned not only in the euchromatic but also in the abnormal (pyknotic or degenerated) nuclei. Following nuclear reconstitution, the nuclear envelope folding is accelerated to induce the usual pattern of AL organization, i.e., budding from the invaginated nuclear envelope. PMID- 2549813 TI - Ultrastructural study of the coiled body and a new inclusion, the "mykaryon," in the nucleus of the adult rat Sertoli cell. AB - Random and serial thin sections of the nucleus in adult rat Sertoli cells were examined by electron microscopy. Besides the previously reported nucleoli and heterochromatin masses, the nuclei contain a coiled body and a new structure, the "mykaryon." The coiled body is 835 nm in maximum diameter. It is composed of distinct elements referred to as "coils." They are 32 nm wide on average and resemble the nucleolar pars fibrosa in their intense staining with heavy metal salts and their composition of narrow filaments. The coiled body is often close to a nucleolus, though no direct contact is established, and it sometimes exists at a distance from the nucleolus. The mykaryon is spherical, 460 nm in maximum diameter, and composed of a tridimensional network of 7-26 nm-wide electron opaque "cords" separated by slightly thinner spaces. It has not been observed in the vicinity of a nucleolus. A literature survey showed no previous mention of a structure similar to the mykaryon. The coiled body and the mykaryon are interpreted as normal constituents of the Sertoli nucleus in the adult rat. PMID- 2549814 TI - Effect of lithium chloride on spermatogenesis, testicular delta 5- 3 beta- and 17 beta-hydroxysteroid dehydrogenase activities in toad (Bufo melanostictus). AB - The purpose of the present study was to show the effects of lithium on testicular activities in the toad. Adult male toads were injected with lithium chloride (200 micrograms/toad) of alternate days for 21 days. At the moment of sacrifice on 22nd day, lithium treated animal showed decreases in testicular weight and Leydig cell nuclear area along with inhibition of spermatogenesis and testicular delta 5 3 beta as well as 17 beta-hydroxysteroid dehydrogenase activities. The results of our present experiment suggest that lithium administration might be associated with significant adverse effects on testicular activity in toad. PMID- 2549815 TI - Cytochrome oxidase and lactate dehydrogenase activities in peroxidized rabbit epididymal spermatozoa. AB - We report a quantitative cytochemical study on cytochrome oxidase and lactate dehydrogenase activities on rabbit epididymal spermatozoa during spontaneous lipid peroxidation. Our data show that during aerobic incubation both in NTP and KTP media the sperm cytochrome oxidase activity undergoes a significant decrease. The lactate dehydrogenase activity shows different cytochemical patterns in comparison between the two media considered. Such activity significantly increases in rabbit spermatozoa suspended in NTP medium from the first until the sixteenth hour of incubation time. At the following times the lactate dehydrogenase activity significantly declines showing yet until the later times of incubation integrated optical density values fairly high. During the whole period of the aerobic incubation, the spermatozoa suspended in medium KTP show lactate dehydrogenase integrated optical density values which not significantly differ from those of the control in spite of an initial enhancement from the first until the thirteenth hour of the experimental treatment. PMID- 2549816 TI - No evidence of the existence of beta adrenoceptors in human sperm using radioligand binding technique. AB - The investigations were carried out on 20 freshly ejaculated and pooled samples of human semen with normozoospermia and motility of 70-80%. In the present study no difference was found between total and non-specific binding to semen membranes using [3H]-dihydroalprenolol (DHA) as a radioligand. Our results suggest that beta adrenoceptors could not be found in human sperm. PMID- 2549817 TI - Occurrence and potential importance of selected peptidases in bull ejaculates. AB - The zinc containing peptidases angiotensin-converting enzyme (ACE), neutral metalloendopeptidase (NEP), and leucine aminopeptidase (LAP) occur in bull ejaculates in high activities. These enzyme activities are in close correlation with some routineously determined semen parameters. These ejaculat parameters are used for quality classification and selection of ejaculates and are according to long term experience in good correlation to the male fertility. Semen quality related correlations could not be found for the dipeptidyl aminopeptidase (DPIV) and the aminopeptidase A (APA). Origin and possible function of the zinc containing peptidases in the semen are examined and discussed. PMID- 2549818 TI - Clinical recovery and train-of-four ratio measured mechanically and electromyographically following atracurium. AB - Indices of clinical recovery were compared with mechanically (adductor pollicis muscle) and electromyographically (first dorsal interosseal muscle) recorded train-of-four (TOF) ratios during antagonism of atracurium blockade in 23 healthy neurolept anesthetized patients. Clinical recovery was evaluated from the ability to lift the head, sustain headlift for 5 or 10 s, protrude the tongue, open the eyes, and the presence of ptosis of the eyelids. In all patients the mechanical TOF ratio was recorded. In 17 patients TOF ratios based on measurements of the potential area and the amplitude of the major negative deflection of the compound EMG response were recorded as well. At each TOF ratio interval of 0.05 from a TOF ratio of 0.5-0.85, the number of patients being able to perform the individual tests was recorded. Further, the mechanical TOF ratio during recovery was compared with the EMG TOF ratios. Headlift could not be sustained for 5 s in any patient at a TOF ratio of 0.5, whether recorded mechanically or by EMG, and TOF ratio had to recover to 0.8 before all patients could sustain headlift for 5 s. All patients could open the eyes and protrude the tongue at a TOF ratio of 0.65, and ptosis remained present during the entire testing period. There was no statistically significant difference between the mechanical and the EMG methods with regard to the TOF ratios at which the tests could be performed. During recovery a linear relationship was found between mechanical TOF ratios and the square root of the EMG TOF ratios.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549819 TI - Depression of myocardial force and stiffness without change in crossbridge kinetics: effects of volatile anesthetics reproduced by nifedipine. AB - The authors examined the effects of nifedipine, a sarcolemmal slow Ca2+ channel blocker, on dynamic stiffness and force of rabbit right ventricular trabeculum and papillary muscle in Ba2+ contracture, in an attempt to reproduce the effects of halothane, enflurane, and isoflurane on a similar preparation as reported by Shibata et al. Once barium contracture force was established, muscle length was perturbed with small amplitude sinusoidal oscillations in the frequency range of 0.1-100 Hz. Nifedipine 1 microM was then added to the superfusate and dynamic stiffness was again measured. Additional barium was used to determine restoration of contracture force to and beyond control levels. Nifedipine produced a significant decrease in contracture force and high-frequency stiffness with no effect on the frequency (fmin) at which stiffness amplitude exhibited a minimum (P less than 0.005). Contracture force and stiffness could be restored by adding additional barium to the nifedipine-treated muscles. These results are similar to those reported by Shibata et al. using volatile anesthetics. Since nifedipine, which acts specifically at the sarcolemmal slow Ca2+ channel, affects contracture force and dynamic stiffness in this preparation in a manner similar to the volatile anesthetics, the authors suggest that the anesthetics studied by Shibata et al. may well exert a significant component of their negative inotropic activity via their action on the sarcolemmal slow Ca2+ channel. PMID- 2549820 TI - Exposure to and precautions for blood and body fluids among workers in the funeral home franchises of Fort Worth, Texas. AB - In 1982 the Centers for Disease Control published a set of recommendations and measures to protect persons working in health care settings or performing mortician services from possible exposure to the human immunodeficiency virus. This study of a number of funeral homes in the Fort Worth area was designed to determine the level of exposure of funeral home workers to blood and other body fluids and also to assess existing protective measures and practices in the industry. Workers in 22 funeral home franchises were surveyed with a predesigned questionnaire. Eighty-five responses from 20 of the 22 establishments were received. All 85 respondents admitted exposure of varying degrees to blood and body fluids. Sixty persons (70%) admitted heavy exposure, that is, frequent splashes. Analysis of the responses showed that 81 of 85 (95.3%) persons consistently wore gloves while performing tasks that might expose them to blood or other body fluids. Of the 60 persons who were heavily exposed, 43 wore long sleeved gowns, 27 wore waterproof aprons, 17 surgical masks, and 15 goggles. The study further revealed that 52.9% (45/85) of the respondents had sustained accidental cuts or puncture wounds on the job. In light of these findings it is important to target educational efforts to persons in this industry to help them minimize their risks of infection with blood and body fluid borne infections. PMID- 2549821 TI - Human immunodeficiency virus testing in health care: Part 1. PMID- 2549822 TI - Management of HIV infection in the hospital. Technical Panel on Infections within Hospitals: American Hospital Association. PMID- 2549823 TI - Natural parvovirus infection in laboratory rabbits. AB - Laboratory rabbits from various commercial and private sources were found to have high serum antibody titers specific for lapine parvovirus (LPV). By both immunofluorescence and hemagglutination inhibition assays, 75% of these sera were positive for LPV. This finding, together with the recovery of LPV from kidneys of neonatal rabbits, suggested that LPV infection is common in commercially available rabbits in the United States. It was concluded that use of infected rabbits could interfere with research in which rabbit cell cultures or in vitro immunologic assays are used. PMID- 2549824 TI - Parathyroid hormone-related protein in adult T-cell leukemia-lymphoma. AB - OBJECTIVE: To determine whether parathyroid hormone-related protein is synthesized and secreted by the tumor cells of patients with adult T-cell leukemia-lymphoma. DESIGN AND PATIENTS: Convenience sample of three patients with adult T-cell leukemia-lymphoma. Two patients developed hypercalcemia, and one patient was normocalcemic. SETTING: Inpatient facilities at two university affiliated medical centers. INTERVENTION: All patients had a lymph node biopsy. In addition, samples of ascitic or pleural fluid, or both, were obtained from these patients. MEASUREMENTS AND RESULTS: Using RNA blot analysis, we showed that parathyroid hormone-related protein (PTHrP) messenger RNAs (mRNAs) were constitutively expressed in the tumor cells from all patients. Cyclic adenosine monophosphate (cAMP) production-stimulating activity, assessed using osteoblast like UMR106 cells, was demonstrated in the pleural and ascitic fluids from the two patients who developed hypercalcemia. The elution profiles of the cAMP production-stimulating activity in the ascitic fluid extracts were very similar to those of the tumor extracts from hypercalcemic nude rats that had been implanted with a human cancer tumor. CONCLUSIONS: Parathyroid hormone-related protein is produced by tumor cells in adult T-cell leukemia-lymphoma, which may be an important factor in the development of hypercalcemia in the patients with this disease. However, the development of hypercalcemia may depend on other factors such as the number of tumor cells, access of the protein into systemic circulation, and the presence of some additional substances. PMID- 2549826 TI - Mediastinal germ cell tumor and acute megakaryoblastic leukemia. PMID- 2549825 TI - Decisions about resuscitation: inequities among patients with different diseases but similar prognoses. AB - STUDY OBJECTIVE: To assess whether decisions about "do-not-resuscitate" (DNR) orders are made equitably in patients with different diseases but similar prognoses. DESIGN: Retrospective cohort study. SETTING: Three teaching hospitals: a university referral center, a county hospital serving a largely indigent population, and a Veterans Administration hospital. PATIENTS: Consecutive patients with any of the four following discharge diagnoses: the acquired immunodeficiency syndrome (AIDS) (100 patients); unresectable non-small-cell lung cancer (51 patients); cirrhosis with esophageal varices (51 patients); and severe congestive heart failure with coronary artery disease (115 patients). MEASUREMENTS AND MAIN RESULTS: Do-not-resuscitate orders were written for 52% of patients with AIDS and 47% of patients with cancer but for only 16% of patients with cirrhosis and 5% of patients with congestive heart failure (P less than 0.0001). Although DNR orders were associated with functional and mental status, reason for admission, and severity of illness, the strong association between DNR orders and disease category persisted after adjustment for these potential confounders by multiple logistic regression. A survey of housestaff showed that DNR orders were discussed more frequently with patients who had AIDS or lung cancer than with patients who had cirrhosis or heart failure, despite an accurate understanding of the generally similar prognoses among the four groups. CONCLUSIONS: Despite relatively similar prognoses, patients with AIDS or lung cancer are much more likely to receive DNR orders than patients with cirrhosis or severe congestive heart failure. This discrepancy cannot be explained by differences in severity of illness among patients or by misunderstandings of prognosis by clinicians. From our data, we cannot determine if patients with cirrhosis or heart failure receive too few DNR orders or if patients with AIDS or lung cancer receive too many. Our findings should encourage physicians to determine the preferences of patients about life-sustaining treatments more equitably. PMID- 2549827 TI - Prostaglandin E2, leukotriene C4, and platelet-activating factor receptor sites in the brain. Binding parameters and pharmacological studies. PMID- 2549828 TI - Antagonistic modulation of S-K+ channel activity by cyclic AMP and arachidonic acid metabolites. Role for two G proteins. PMID- 2549829 TI - The role of arachidonic acid and oxygen radical metabolites in the pathogenesis of vasogenic brain edema and astrocytic swelling. PMID- 2549831 TI - The masseteric (jaw closure) reflex: a simple mammalian brainstem system for studying neurochemical modulation. PMID- 2549830 TI - Ischemic injury in the brain. Role of oxygen radical-mediated processes. PMID- 2549832 TI - Regulation of c-fos messenger ribonucleic acid by fibroblast growth factor in cultured Sertoli cells. AB - The data reported here demonstrate that basic FGF and Sertoli-cell-derived FGF have rapid stimulatory effects on c-fos mRNA levels. Basic FGF appears to utilize a signal transduction pathway that is distinct from that used by FSH and serum but similar in its potency and transiency. This is consistent with other reports in the literature on FGFs mechanism of action. For example, a monoclonal antibody to phosphatidyl 4,5-biphosphate will block the effects of PDGF on thymidine incorporation into NIH 3T3 cells but has no effect on basic FGF. Our data support the emerging possibility of a novel pathway mediating FGF actions. A similar precedent has been established for serotonin-induced DNA synthesis in smooth muscle cells. The rat Sertoli cell, with both FSH and FGF rapidly inducing c-fos, is an excellent model system for addressing the mechanism of action of basic FGF, the specificity of the c-fos response and the role of FGF in the reproductive system. Stimulation of c-fos mRNA by basic FGF in the cultured Sertoli cell presents questions regarding the role of FGF and c-fos in the male reproductive system. Basic FGF has been shown to stimulate cell division and plasminogen activator activity in cultured immature porcine Sertoli cells. Plasminogen activator may play a critical role in the tissue remodeling required for spermiogenesis. Interestingly, fos has been shown to be expressed preferentially in pachytene spermatocytes in the mouse. These observations taken together with the finding that basic FGF mRNA levels in Xenopus oocytes are markedly elevated, suggests that basic FGF may be important for Sertoli cell function, spermatogonial cell proliferation and subsequent meiotic and sperm maturational events. PMID- 2549833 TI - Oxytocin in the testis: what, where, and why? PMID- 2549834 TI - Sertoli cell-germ cell communication. PMID- 2549835 TI - Developmental expression of the sperm receptor of the mouse zona pellucida. PMID- 2549836 TI - Mechanisms of signal transduction in mouse spermatozoa. AB - Sperm from all species studied thus far contain G-proteins. The presence of such signal-transducing proteins in these cells suggests that the regulation of sperm function might have control elements that are similar to ligand:receptor:G protein:second messenger systems common to many somatic cells. This hypothesis is supported by experiments that demonstrate a potential intermediary role for the mouse sperm Gi-like protein in the acrosome reaction induced by ZP3. The specific function of this Gi-like protein in this important physiologic event is not known at this time, although possible roles in regulating ionic movements, cyclic nucleotide metabolism, and polyphosphoinositide turnover are possible candidates. Studies directed at the localization and biochemical identity of the mouse sperm Gi-like protein, as well as the nature of the second messenger system(s) modulated by this protein, are in progress and should help to delineate the sequence of events involved in some of the early steps of sperm-ZP interaction. PMID- 2549838 TI - Surgical treatment of the penoscrotal Paget's disease. AB - Data from six cases of penoscrotal Paget's disease were collected. Erythematous, indurated, and pruritic skin lesions that failed to respond to the topical therapy were the main complaints. Symptoms had been observed for three to seven years before correct diagnosis. Extensive diagnostic studies revealed negative findings of subjacent adenocarcinoma, but 1 patient had concurrent skin cancer of nasal ala. In treating penoscrotal Paget's disease, our policy is to (1) resect 3 cm or more skin beyond the margin of the cutaneous lesion, (2) resect deeply to the subcutaneous fat, (3) use intraoperative frozen sections to confirm the free margin and (4) reconstruct the wound with the local iliac flap. No local recurrence was noted during the follow-up period of two to seven years. The iliac flap is thin and pliable; it precluded scar contracture and painful penile erection, which are the common sequelae of free skin graft in the penoscrotal area. Patients enjoyed normal sexual activity after surgery. PMID- 2549837 TI - Regulation of the differentiated functions of Leydig tumor cells by epidermal growth factor. AB - The three effects of mEGF on MA-10 Leydig tumor cells that have been discussed here are summarized in TABLE 7. The earliest effect of mEGF on MA-10 cells can be detected within 5 min of addition of mEGF and it lasts for about 60 min. During this time mEGF transiently attenuates hCG-stimulated adenylate cyclase activity. Although the magnitude of this effect is small, it can be correlated with a transient attenuation of the hCG-provoked increase in steroid synthesis. At longer times (i.e., 1-8 h) mEGF activates steroid synthesis by a "cAMP independent pathway" and it potentiates (in a synergistic fashion) the activation of steroidogenesis by hCG, other compounds that activate adenylate cyclase activity, and cAMP analogues. At even longer times (i.e., 8-48 h) mEGF down regulates the LH/CG receptors and by doing so, limits the steroidogenic response of the cells to hCG. From a biochemical point of view, our data provide an excellent example of those actions of growth factors that are unrelated to the control of cell multiplication, and of the complexity involved even when dealing with a single cell type and a single growth factor. Admittedly we know very little about the molecular basis of the phenomena described herein. Current work in our laboratory, however, is aimed at filling this gap. Among all the questions that we can address, we believe that it is particularly important to characterize the intracellular signaling system(s) activated by mEGF and to determine if a single signaling system is responsible for the diverse biological actions of mEGF in MA-10 cells. From a physiological point of view, our data may also prove important to the understanding of the regulation of testicular functions. There is increasing evidence for the production of EGF (or related peptides such as transforming growth factor alpha) in several tissues, including the testes and ovaries. These findings, together with the results summarized here suggest that EGF (or related peptides) act within the testes in a paracrine, or autocrine fashion and that they may have important modulatory effects on the activation of Leydig cell steroidogenesis by gonadotropins. PMID- 2549839 TI - The surgical conservation of the neuropathic foot. AB - Basic surgical principles applied when caring for neuropathic limbs can result in the maintenance or restoration of a useful ulcer-free limb. It is possible to help many patients with neuropathy to become ulcer-free and to remain ulcer-free and mobile, with surgical procedures. Recommended methods of management are briefly outlined. These include the debridement of the osteomyelitic, metatarsal head in order to save the adjacent toe, removal of bony irregularities that predispose to ulceration, and the use of wedge osteotomies and arthrodeses to improve the functional shape of the affected foot. The emphasis is on the removal of high pressure points from the weight-bearing surface and to increase the total area available for weight-bearing. Adequate rest and protection are essential, and include the use of splints or total contact plaster casts in all cases of ulceration of weight-bearing surfaces. All patients with reduced sensory perception should learn daily self-examination and care to reduce the chances of recurrent ulceration. Healing after surgical reconstruction will occur, and the healed tissues, if adequately cared for, will maintain their integrity for years. PMID- 2549840 TI - Unilateral facet joint hypertrophy causing nerve root irritation. PMID- 2549842 TI - Incidence of primary liver cancer in Singapore, 1968-1982. AB - Based on data collected by the population-based Singapore Cancer Registry over the period 1968 to 1982, baseline epidemiological characteristics and incidence trends of primary liver cancer were described. This will facilitate the interpretation of future trends, especially in the light of new interventions such as hepatitis B immunisation. The primary liver cancer incidence is four times higher in males than in females, with the incidence peaked in the seventh decade. The incidence rate was higher in the Chinese than in Malays and Indians and marginally higher among foreign born than Singapore born Chinese. A general declining trend in liver cancer incidence was especially notable in the local born Chinese. Misclassification of metastatic carcinomas in the earlier years of cancer registration may have contributed to the initial higher incidence. Definitive decrease in incidence as a result of hepatitis immunisation will only be seen in another two to three decades. PMID- 2549841 TI - Agranulocytosis and anaemia induced by sulfametopyrazine in a sulfametopyrazine trimethoprim combination. AB - We report a case of prolonged fever, agranulocytosis, and anaemia associated with the long acting sulphametopyrazine-trimethoprim combination (Kelfiprim). A woman of 23 years took an overdose of 13 tablets over five days for presumed cystitis. One day after the last dose the patient developed fever and a generalised rash. The fever persisted and her previously normal leukocyte count decreased to 1.8 x 10(9)/1. After treatment with paracetamol the fever settled briefly, and then recurred for another 16 days. A later peripheral blood leukocyte count of 0.77 x 10(9)/1, haemoglobin of 10.8 g/dl, and a hypocellular bone marrow with depressed granulopoiesis and haemopoiesis suggested marrow suppression induced by sulfametopyrazine. Since the IgM antibody against the Epstein-Barr virus capsid antigen was detected, the adverse drug reaction might have been aggravated by this virus. The case highlights the risk of severe haematological adverse reactions associated with sulphonamide treatment, and argues for the use of trimethoprim alone for uncomplicated cystitis. PMID- 2549843 TI - Myoclonic epilepsy and ragged-red fibers with cytochrome oxidase deficiency: neuropathology, biochemistry, and molecular genetics. AB - A 36-year-old man with myoclonic epilepsy and ragged-red fibers (MERRF) died after more than 18 years of follow-up study. He was 1 of 3 affected siblings and the offspring of an affected mother, suggesting maternal transmission. At autopsy, there was neuronal loss and gliosis in the dentate nucleus of the cerebellum and in the inferior olivary nucleus. Skeletal muscle showed ragged-red fibers, and paracrystalline inclusions in mitochondria by electron microscopy. Biochemical analysis showed a generalized partial defect of cytochrome c oxidase (COX) in mitochondria isolated from all tissues, including brain, heart, skeletal muscle, kidney, and liver. The Michaelis constant (Km) for cytochrome c was abnormally low, suggesting a defect of the mitochondrially encoded subunit II of COX. Immunological studies (enzyme-linked immunosorbent assay, dot-blot, Western blot, and immunohistochemistry) showed that the holoenzyme was decreased but subunit II was decreased more than the holocomplex or the nuclearly encoded subunit IV. However, Northern and Southern blots showed that the gene for subunit II, as well as the genes for subunits I, III, IV, and VIII, were of normal size and were normally transcribed. A point mutation or a small deletion of mitochondrial DNA, probably affecting the COX-II gene, may be responsible for the COX deficiency in this case of MERRF. PMID- 2549844 TI - Lipid-containing cells in brains of normal and hypoxic infant monkeys: a quantitative and ultrastructural study. AB - The significance of lipid-containing cells found at autopsy in the white matter of infant brains is controversial, particularly with respect to their postulated role as markers of the "sudden infant death syndrome." To determine whether such cells are indicative of prior nonlethal hypoxic insult, we quantitated them in the brains of control infant monkeys and in two groups of infant monkeys that were subjected to 30 minutes of hypoxic insult. One group consisting of monkeys that died less than 48 hours after the hypoxia, and the other of those that survived 7 to 13 days following the insult. The quantification of lipid containing cells was undertaken in frozen brain sections stained with Oil red O; sections of brains from 4 perfusion-fixed animals were evaluated by electron microscopy. Lipid-containing cells were found in the corpus callosum, in the septum, and in periventricular white matter in both posthypoxic and control animals. There was a relationship between numbers of lipid-containing cells and the age of the animal; animals with large numbers were less than 28 days old. Decreases in numbers of lipid-containing cells correlated with advancing myelination as well as with age. Electron microscopic evaluation revealed lipid droplets in the cytoplasm of cells with irregularly shaped nuclei, densely clumped chromatin, occasional microtubules, and narrow cytoplasmic processes. We suggest that lipid-containing cells in the white matter of the brains of infants are related to age and to maturational factors and, in the absence of other pathologic signs, are not related to prior hypoxic injury. PMID- 2549845 TI - The pedunculopontine nucleus in Parkinson's disease. AB - This study demonstrated a significant loss of neurons within the lateral part of the pedunculopontine nucleus pars compacta in individuals with idiopathic Parkinson's disease and in individuals with combined Parkinson's and Alzheimer's disease. We also examined the extent of neuronal loss within the substantia nigra pars compacta, locus ceruleus, dorsal raphe nucleus, and nucleus basalis of Meynert. The number of pedunculopontine nucleus pars compacta neurons in the patients with Parkinson's or Parkinson's and Alzheimer's disease was reduced (average, 40%) in comparison with the number in control subjects or patients with Alzheimer's disease (p less than 0.01). This finding correlated significantly with the extent of loss of substantia nigra pars compacta neurons (p less than 0.01). PMID- 2549846 TI - Dementia in Parkinson's disease is related to neuronal loss in the medial substantia nigra. AB - Regional neuronal loss in the substantia nigra was studied in relation to extrapyramidal symptoms and dementia in 12 patients with idiopathic Parkinson's disease (PD) and in 18 control subjects. Four areas of the right substantia nigra were investigated at the level of the superior colliculus and caudal red nucleus. In Parkinson's disease, the percentages of neurons, from the medial to the lateral part of the substantia nigra, were reduced to 49%, 31%, 41%, and 25% of the control values. The number of neurons in the lateral part showed a negative correlation with the severity of rigidity and hypokinesia, whereas tremor was less noticeable in patients with few neurons. The degree of dementia of the patients had a significant correlation only with neuronal loss in the medial part of the substantia nigra, suggesting, in view of the topographical organization of the neurons in the substantia nigra, that intact projections to the caudate nucleus and limbic and cortical areas are a prerequisite for normal cognitive functioning and that their dysfunction leads to clinical dementia. PMID- 2549847 TI - Cerebrospinal fluid content of diazepam binding inhibitor in chronic hepatic encephalopathy. AB - The neuropeptide diazepam binding inhibitor (DBI) is an endogeneous allosteric modulator of gamma-aminobutyric acid (GABA) receptors at the benzodiazepine recognition site. Recent theories on the neurochemical cause for hepatic encephalopathy have implicated activation of inhibitory neurotransmitter GABA systems. In 20 patients with hepatic disease, blood and cerebrospinal fluid (CSF) levels of ammonia and amino acids were measured. As in previous studies there was a selective elevation of CSF amino acids as well as a correlation between CSF glutamine levels and encephalopathy. CSF DBI levels were maximally elevated 5 fold in patients with hepatic encephalopathy, but they were normal in those patients with liver disease not associated with changes in mental status and in patients with nonhepatic encephalopathy. Levels of DBI correlated with the clinical staging of hepatic encephalopathy. These data suggest that DBI may participate in the modulation of cerebral function in hepatic encephalopathy. PMID- 2549848 TI - The effect of MK-801 on kindled seizures: implications for use and limitations as an antiepileptic drug. AB - MK-801 is a new drug that produces a noncompetitive blockade at the subclass of glutamate receptors activated by N-methyl-D-asparate (NMDA). The antiepileptic properties of MK-801 were studied using kindled seizures as a model of complex partial seizures with secondary generalization. A test protocol was employed that allowed: (1) examination of the efficacy of MK-801 against several parameters that gauge different aspects of epileptogenesis; (2) determination of the time action profile of these effects; and (3) examination of the toxicity of MK-801 in animals experiencing seizures. The drug was found to be potent against the spread of seizures but less effective against parameters linked to partial seizures. At the higher doses of the drug required to truncate hippocampal afterdischarges, considerable neurotoxicity was encountered. In addition, the antiepileptic effects of MK-801 showed a use dependence so that, at a given time after the drug was administered, a greater suppression of seizures was noted if there had been preceding seizures in the presence of the drug than if there had not been. These findings indicate that there may be limitations to the clinical utility of MK-801 as an antiepileptic agent and that the drug may provide greatest benefit when used for the suppression of seizure generalization and when seizures are closely spaced. PMID- 2549849 TI - Tomaculous neuropathy presenting as acute recurrent polyneuropathy. AB - Tomaculous neuropathy (TN) is classically associated with the inherited, recurrent focal neuropathies. We report a case of TN manifesting as an acute recurrent polyneuropathy. A 28-year-old woman had 2 episodes of acute, ascending, symmetrical sensorimotor deficit. Teased nerve-fiber preparation confirmed the presence of TN. Extensive investigations failed to reveal other cause for her symptoms. We believe that this case is unique and broadens the clinical spectrum of TN. PMID- 2549850 TI - Evidence for a novel picornavirus in human dermatomyositis. AB - We prepared RNA probes from cloned segments of human and murine enteroviruses (EVs) for in situ hybridization of skeletal muscle biopsies from patients with dermatomyositis (DM), polymyositis, other inflammatory myopathies, and noninflammatory muscle diseases, and from normal control subjects. A probe derived from Theiler's murine encephalomyelitis virus (TMEV) detected viral RNA within mononuclear cells of the interstitial connective tissue in 3 of 5 patients with adult-onset DM. None of these patients showed positive hybridization to probes derived from human EVs (poliovirus type 1 and Coxsackie virus B3) applied to subjacent sections of the same biopsies. The remaining 2 adult DM patients, 4 patients with childhood-onset DM, and 24 non-DM patients did not react with either TMEV or human enterovirus probes. Histochemical stains for esterase and immunoperoxidase stains for Mac-1 antigen in the 3 DM patients who reacted positively revealed positive cells in the same distribution as, but in far greater number than, those positive by in situ hybridization. Immunoperoxidase staining for HLA-DR antigens revealed positive cells in the same distribution and number as were seen with the TMEV probe. We conclude that an EV-like agent, more closely related to TMEV than to human EVs, may be associated with DM and that this agent is probably localized within muscle macrophages that express class II major histocompatibility complex antigens. PMID- 2549851 TI - Isolation of herpes simplex virus type 1 during first attack of multiple sclerosis. AB - Herpes simplex virus type 1 was isolated from the cerebrospinal fluid of a patient during the first attack of multiple sclerosis. This is the first virus to be isolated from the central nervous system of a living patient with MS. The virus was identified as herpes simplex virus type 1 by restriction endonuclease analysis and by an enzyme immunoassay using monoclonal antibodies. Antibodies against type 1 but not type 2 were detected in consecutive samples of serum and cerebrospinal fluid. The patient has since entered a progressive phase of multiple sclerosis. The isolated type 1 strain might be of pathogenetic relevance to the development of multiple sclerosis in this patient. PMID- 2549852 TI - The protein kinase C family: heterogeneity and its implications. PMID- 2549853 TI - DNA topoisomerase poisons as antitumor drugs. PMID- 2549854 TI - Quinoproteins, enzymes with pyrrolo-quinoline quinone as cofactor. PMID- 2549855 TI - Hemopoietic cell growth factors and their receptors. PMID- 2549856 TI - The structure and regulation of protein phosphatases. AB - Four major serine/threonine-specific protein phosphatase catalytic subunits are present in the cytoplasm of animal cells. Three of these enzymes, PP-1, PP-2A, and PP-2B, are members of the same gene family, while PP-2C appears to be distinct. PP-1, PP-2A, and PP-2B are complexed to other subunits in vivo, whereas PP-2C has only been isolated as a monomeric protein. PP-1, PP-2A, and PP-2C have broad and overlapping specificities in vitro, and account for virtually all measurable activity in tissue extracts toward a variety of phosphoproteins that regulate metabolism, muscle contractility, and other processes. Their precise functions in vivo are unknown, although important clues to the physiological roles of PP-1 and PP-2A are provided by the effects of okadaic acid and by the subcellular localization of PP-1. The active forms of PP-1 are largely particulate, and their association with subcellular structures is mediated by "targetting subunits" that direct PP-1 to particular locations, enhance its activity toward certain substrates, and confer important regulatory properties upon it. This concept is best established for the glycogen-bound enzymes in skeletal muscle and liver (PP-1G) and the myofibrillar form (PP-1M) in skeletal muscle. The activities of PP-1 and PP-2B are controlled by the second messengers cyclic AMP and calcium. The activity of PP-2B is dependent on calcium and calmodulin, while PP-1 is controlled in a variety of ways that depend on the form of the enzyme and the tissue. PP-1 can be inhibited by cyclic AMP in a variety of cells through the A-kinase-catalyzed phosphorylation of inhibitor-1 and its isoforms. Phosphorylation of the glycogen-binding subunit of PP-1G by A-kinase promotes translocation of the catalytic subunit from glycogen particles to cytosol in skeletal muscle, inhibiting the dephosphorylation of glycogen metabolizing enzymes. Allosteric inhibition of hepatic PP-1G by phosphorylase a occurs in response to signals that elevate cyclic AMP or calcium, and prevents the activation of glycogen synthase in liver. PP-1 can also be activated indirectly by calcium through the ability of PP-2B to dephosphorylate inhibitor 1. This control mechanism may operate in dopaminoceptive neurones of the brain and other cells. The inactive cytosolic form of PP-1 (PP-1I) can be activated in vitro through the glycogen synthase kinase-3-catalyzed phosphorylation of its inhibitory subunit (inhibitor-2), but the physiological significance is unclear.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2549858 TI - Animal virus DNA replication. PMID- 2549857 TI - The heparin-binding (fibroblast) growth factor family of proteins. PMID- 2549859 TI - Molecular basis of fertilization. PMID- 2549860 TI - Selected papers from the 1st International Symposium on PQQ and Quinoproteins. Delft, The Nethlerlands, 5-7 September 1988. PMID- 2549861 TI - Mutants of Methylobacterium organophilum unable to synthesize PQQ. AB - The phenotype of mutants unable to synthesize PQQ is analyzed for different categories of methylotrophic bacteria. The advantages offered by strains dissimilating methylamine through methylated amino-acids are discussed. In M.organophilum, 40% of the mutants unable to grow in methanol medium but with normal methylamine utilization, were affected in PQQ metabolism. The genetic properties of M. organophilum useful to study PQQ mutants are discussed, mainly the use of pSUP106 to create insertion mutations in the bacterial chromosome and to replace wild-type genes by modified genes. An example is given of the possibility to create R' plasmids containing large fragments of M.organophilum DNA. Some physiological properties of a PQQ mutant are described, regarding growth kinetics, PQQ uptake and accumulation. PMID- 2549862 TI - Quinoproteins in C1-dissimilation by bacteria. PMID- 2549863 TI - PQQ and quinoproteins: an important novel field in enzymology. PMID- 2549864 TI - Physiological significance and bioenergetic aspects of glucose dehydrogenase. AB - The regulation of the PQQ-linked glucose dehydrogenase in different organisms is reviewed. It is concluded that this enzyme functions as an auxiliary energy generating mechanism, because it is maximally synthesized under conditions of energy stress. It is now definitively established that the oxidation of glucose to gluconate generates metabolically useful energy. The magnitude of the contribution of the oxidation of glucose to gluconate via this enzyme to the growth yield of organisms such as Acinetobacter calcoaceticus is not yet clear. PMID- 2549865 TI - Quinoprotein D-glucose dehydrogenases in Acinetobacter calcoaceticus LMD 79.41: purification and characterization of the membrane-bound enzyme distinct from the soluble enzyme. AB - Acinetobacter calcoaceticus is known to contain soluble and membrane-bound quinoprotein D-glucose dehydrogenases while other oxidative bacteria such as Pseudomonas or Gluconobacter contain only membrane-bound enzyme. The two different forms were believed to be the same enzyme or interconvertible. Present results show that the two different forms of glucose dehydrogenase are distinct from each other in their enzymatic and immunological properties as well as in their molecular size. The soluble and membrane-bound glucose dehydrogenases were separated after French press-disruption by repeated ultracentrifugation, and then purified to nearly homogeneous state. The soluble enzyme was a polypeptide of 55 Kdaltons, while the membrane-bound enzyme was a polypeptide of 83 Kdaltons which is mainly monomeric in detergent solution. Both enzymes showed different enzymatic properties including substrate specificity, optimum pH, kinetics for glucose, and reactivity for ubiquinone-homologues. Furthermore, the two enzymes could be distinguished immunochemically; the membrane-bound enzyme is cross reactive with an antibody raised against membrane-bound enzyme purified from Pseudomonas but not with antibody elicited against the soluble enzyme, while the soluble enzyme is not cross-reactive with the antibody of membrane-bound enzyme. Data also suggest that the membrane-bound enzyme functions by linking to the respiratory chain via ubiquinone though the function of the soluble enzyme remains unclear. PMID- 2549866 TI - Genes involved in the biosynthesis of PQQ from Acinetobacter calcoaceticus. AB - From a gene bank of the Acinetobacter calcoaceticus genome a plasmid was isolated that complements four different classes of PQQ- mutants. Subclones of this plasmid revealed that the four corresponding PQQ genes are located on a fragment of 5 kilobases. The nucleotide sequence of this 5 kb fragment was determined and by means of Tn5 insertion mutants the reading frames of the PQQ genes could be identified. Three of the PQQ genes code for proteins of Mr 29700 (gene I), Mr 10800 (gene II) and Mr 43600 (gene III) respectively. In the DNA region where gene IV was mapped however the largest possible reading frame encodes for a polypeptide of only 24 amino acids. A possible role for this small polypeptide will be discussed. Finally we show that expression of the four PQQ genes in Acinetobacter 1woffi and Escherichia coli lead to the synthesis of the coenzyme in these organisms. PMID- 2549867 TI - PQQ: biosynthetic studies in Methylobacterium AM1 and Hyphomicrobium X using specific 13C labeling and NMR. AB - Using 13C labeling and NMR spectroscopy we have determined biosynthetic precursors of pyrroloquinoline quinone (PQQ) in two closely related serine-type methylotrophs, Methylobacterium AM1 and Hyphomicrobium X. Analysis of the 13C labeling data revealed that PQQ is constructed from two amino acids: the portion containing N-6,C-7, 8, 9 and the two carboxylic acid groups, C-7' and 9', is derived-intact -from glutamate. The remaining portion is derived from tyrosine; the phenol side chain provides the six carbons of the ring containing the orthoquinone, whereas internal cyclization of the amino acid backbone forms the pyrrole-2-carboxylic acid moiety. This is analogous to the cyclization of dopaquinone to form dopachrome. Dopaquinone is a product of the oxidation of tyrosine (via dopa) in reactions catalyzed by monophenol monooxygenase (EC 1.14.18.1). Starting with tyrosine and glutamate, we will discuss possible biosynthetic routes to PQQ. PMID- 2549868 TI - Malignant glioma. A case study. PMID- 2549869 TI - Hyperoxia and xanthine dehydrogenase/oxidase activities in rat lung and heart. AB - Cell injury from hyperoxia is associated with increased formation of superoxide radicals (O2-). One potential source for O2- radicals is the reduction of molecular O2 catalyzed by xanthine oxidase (XO). Physiologically, this reaction occurs at a relatively low rate, because the native form of the enzyme is xanthine dehydrogenase (XD) which produces NADH instead of O2-. Reports of accelerated conversion of XD to XO, and increased formation of O2- formation in ischemia-reperfusion injury, led us to examine whether hyperoxia, which is known to increase O2- radical formation, is associated with increased lung XO activity, and accelerated conversion of XD to XO. We exposed 3-month-old rats either to greater than 98% O2 or room air. After 48 h, we sacrificed the rats and measured XD and XO activities and uric acid contents of the lungs. We also measured the activities of the two enzymes in the heart as a control organ. We found that the activity of XD was not altered significantly by hyperoxia in rat lungs or hearts, but XO activity was markedly lower in the lung, whether expressed per whole organ or per milligram protein, and remained unchanged in the heart. Lung uric acid content was also significantly lower with hyperoxia. The decrease in lung XO activity may reflect inactivation of the enzyme by reactive O2 metabolites, possibly as a negative feedback mechanism. The concomitant decrease in uric acid content suggests either decreased production mediated by XO due to its inactivation or greater utilization of uric acid as an antioxidant. We examined these postulates in vitro using a xanthine/xanthine oxidase system and found that H2O2, but not uric acid, has an inhibitory effect on O2- formation in the system. We therefore conclude that hyperoxia is not associated with increased conversion of XD to XO, and that the exact contribution of XO to hyperoxic lung injury in vivo remains unclear. PMID- 2549870 TI - Inhibition of inorganic pyrophosphatase of animal mitochondria by calcium. AB - Calcium ion is an uncompetitive inhibitor of the inorganic pyrophosphatases of bovine heart and rat liver mitochondria with respect to substrate MgPPi at pH 8.5 and a non-competitive inhibitor of the former enzyme at pH 7.2. The concentration of Ca2+ required to decrease the maximal velocities for both enzymes at pH 8.5, 0.4 mM Mg2+ was about 10 microM. The inhibition results from the binding of two Ca2+ ions to both free enzymes and their complexes with the substrate. The results suggest that Ca2+ regulates pyrophosphatase activity and hence PPi level in mammalian mitochondria. PMID- 2549871 TI - Kinetic models for the action of cytosolic and mitochondrial inorganic pyrophosphatases of rat liver. AB - Initial rates of PPi hydrolysis by cytosolic and mitochondrial inorganic pyrophosphatases of rat liver have been measured in the presence of 0.2-100 microM MgPPi and 0.01-50 mM Mg2+ at pH 7.2 to 9.3. The apparently simplest model consistent with the data for both enzymes implies that they bind substrate, in the form of MgPPi, and three Mg2+ ions, of which two are absolutely required for activity. The third metal ion facilitates substrate binding but decreases maximal velocity for the cytosolic enzyme, while substrate binding is only modulated for the mitochondrial enzyme. The model is also applicable to bovine heart mitochondrial pyrophosphatases. The active form of the substrate for the cytosolic pyrophosphatase is MgP2O7(-2); the catalytic and metal-binding steps require a protonated group with pKa = 9.2 and an unprotonated group with pKa = 8.8, respectively. The results indicate that the mitochondrial pyrophosphatase is more sensitive to variations of Mg2+ concentration in rat liver cells than is the cytosolic one. PMID- 2549872 TI - Allosteric regulation of yeast inorganic pyrophosphatase by substrate. AB - Kinetic and binding studies of yeast inorganic pyrophosphatase (EC 3.6.1.1) revealed a regulatory PPi-binding site. Rate vs substrate concentration dependencies were markedly nonhyperbolic in the range of 0.1-150 microM MgPPi at fixed Mg2+ levels of 0.05-10 mM provided that the enzyme had been preequilibrated with Mg2+. Imidodiphosphate, hydroxymethylenebisphosphonate, and phosphate eliminated the deviations from the Michaelis-Menten kinetics and inhibited PPi hydrolysis in a manner consistent with their binding at both active and regulatory sites. The results agreed with a model in which binding of uncomplexed PPi at the regulatory site markedly increases enzyme affinity for the activating Mg2+ ion. Ultrafiltration studies revealed the binding of at least 3 mol of the inhibitory hydroxymethylenebisphosphonate and of 2 mol of noninhibitory methylenebisphosphonate per mole of the dimeric enzyme. PMID- 2549873 TI - Synergism between protein kinase C activator and fatty acids in stimulating superoxide anion production in guinea pig polymorphonuclear leukocytes. AB - Treatment of guinea pig polymorphonuclear leukocytes (PMNL) with various fatty acids elicited superoxide anion (O2-) production and an increase in intracellular Ca2+ [( Ca2+]i). Both responses, however, were seldom observed when PMNL were treated at lower concentrations. But, simultaneous addition of 1-oleoyl-2 acetylglycerol (OAG), a protein kinase C activator, caused an increase in O2- production even at the lower concentrations of fatty acids. In contrast to the synergism in O2- production, [Ca2+]i remained at almost the basal level irrespective of the presence of OAG. Among saturated fatty acids, those with carbon numbers of 14 to 18 were most effective in stimulating O2- production in combination with OAG. Unsaturated fatty acids with a carbon number of 18 were almost equally effective irrespective of the number of double bonds. PMID- 2549874 TI - Purification and properties of cyclic-3',5'-GMP-dependent protein kinase from the nematode Ascaris suum. AB - A cyclic-3',5'-GMP-dependent protein kinase was purified 7400-fold from the reproductive tract of female ascarids to a specific activity of 718 nmol min-1 mg 1 (histone as substrate). The yield of the preparation was 25%. The enzyme protein obtained was homogeneous as judged by isoelectrofocusing and polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The native enzyme behaved as a dimer of two 82-kDa subunits in gel permeation chromatography on Superose 12. The protein kinase was inactive in the absence of cyclic purine nucleotides. Half-maximum velocity was obtained in the presence of 18 nM cGMP, whereas 400-fold higher concentrations of cAMP were required for the same activity. The enzyme underwent autophosphorylation in first-order kinetics (rate constant 0.054 min-1), leading to maximum incorporation of 0.96 phosphate per subunit. The autophosphorylation led to a 4-fold increase in Vmax, while the Km remained almost unchanged. In an extract from the reproductive tract, cGMP stimulated phosphorylation was primarily observed in five proteins (molecular masses of 66, 60, 43, 30, and 25 kDa). These proteins also incorporated phosphate when isolated reproductive tracts were incubated in the presence of [32P]phosphate. The phosphate content in cellular proteins was enhanced when the incubation was performed in the presence of 10(-4) M of either octyl-cAMP or octyl-cGMP. In addition to the proteins mentioned above, however, six more electrophoretic bands containing radioactive phosphate were identified after in situ labeling of reproductive tracts with radioactive phosphate. PMID- 2549875 TI - The glycosphingolipid composition of the human hepatoma cell line,Hep-G2. AB - The origin of plasma glycosphingolipids in normal individuals and the mechanisms by which tumor-associated glycosphingolipid antigens enter the plasma in patients with cancer are largely unknown. The Hep-G2 human hepatoma cell line retains many of the characteristics of differentiated hepatocytes including the ability to synthesize and secrete lipoproteins. Preliminary results indicated that newly synthesized Hep-G2 cell glycosphingolipids are coupled to the secreted lipoproteins. This suggests that this cell line may offer an interesting model for studying glycosphingolipid secretion, transfer, and shedding. We now report on the chemical and immunological characterization of Hep-G2 cell glycosphingolipids. Five major glycosphingolipids were purified and biochemically characterized: glycosylceramide, lactosyl ceramide, ceramide trihexoside, ganglioside GM3, and lactosyl sulfatide. Four additional minor components (3 fucosyl-lactosamine containing glycolipids, asialo GM2, galactosylgloboside, and ganglioside GM1) were identified using a combination of exoglycosidase digestion and immunostaining of thin-layer chromatography plates with specific carbohydrate binding proteins. This demonstrates that although this cell line synthesizes a limited number of major glycosphingolipids, it retains the ability to produce at least small amounts of structures in the lactoneo, globo, and ganglio series of glycosphingolipids. These studies show that it will be possible to investigate the mechanisms of secretion by Hep-G2 cells of different classes of these molecules such as neutral glycosphingolipids, gangliosides, and sulfatides. PMID- 2549877 TI - Nick translation of lambda phage DNA with a deoxycytidine analog spin labeled in the 5 position. AB - The synthesis and properties of a novel C(5)-spin-labeled 2'-deoxycytidine 5' triphosphate which serves as a suitable substrate for the template-directed enzyme Escherichia coli DNA polymerase I are reported. The spin label is readily incorporated into lambda phage DNA by nick translation where it reports the characteristic local base motion for double- and single-stranded DNA as determined by electron spin resonance. The high-frequency deoxycytidine motion is similar to the previously reported thymidine motion in double-stranded lambda phage DNA. PMID- 2549876 TI - Activation of 5-lipoxygenase by guanosine 5'-O-(3-thiotriphosphate) and other nucleoside phosphorothioates: redox properties of thionucleotide analogs. AB - The stable nucleotide analog guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) was found to be a very potent activator of 5-lipoxygenase in cell-free preparations from rat polymorphonuclear (PMN) leukocytes, causing a 10-fold stimulation of arachidonic acid oxidation at concentrations as low as 0.5-1 microM. The enhancement of enzyme activity was not directly related to G protein activation since the effect of GTP gamma S could not be abolished by GDP nor replaced by GTP or guanylyl-imidodiphosphate (up to 100 microM). Furthermore, other phosphorothioate analogs, such as guanosine 5'-O-(2-thiodiphosphate), adenosine 5'-O-(3-thiotriphosphate), adenosine 5'-O-(2-thiodiphosphate), and adenosine 5'-O-thiomonophosphate all stimulated 5-lipoxygenase activity at concentrations of 10 microM or lower. This effect could not be detected with any of the corresponding nucleoside phosphate derivatives. The stimulation of 5 lipoxygenase activity by nucleoside phosphorothioates was observed under conditions where the reaction is highly dependent on exogenous hydroperoxides, such as in the presence of beta-mercaptoethanol or using enzyme preparations pretreated with sodium borohydride or glutathione peroxidase. GTP gamma S stimulated arachidonic acid oxidation by 5-lipoxygenase to the same extent as the activating hydroperoxides but had no effect on the reaction measured in the presence of optimal concentrations of 13-hydroperoxyoctadecadienoic acid (1-5 microM). Finally, sodium thiophosphate, but not sodium phosphate, markedly stimulated 5-lipoxygenase activity with properties similar to those of GTP gamma S. These results indicate that GTP gamma S and other phosphorothioate derivatives have redox properties that can contribute to increase 5-lipoxygenase activity by replacing the effect of hydroperoxides. PMID- 2549878 TI - [Recent advances in pathology and biology of small cell lung cancer]. AB - Small cell lung cancer (SCLC) is histologically simple and looks undifferentiated, but possesses cytoplasmic dense-cored granules resembling neuroendocrine granules, and frequently produces amine and peptide hormones, occasionally presenting related symptoms. Among these bioactive substances, gastrin releasing peptide (GRP) is most important, which is known as autocrine growth factor and one of the useful monitoring markers for SCLC together with neuron-specific enolase. Aromatic L-amino acid decarboxylase is another important enzyme in SCLC. Abnormality of myc family oncogenes is occasionally noted in SCLC, which appears related to proliferative activity of the tumor rather than development. Deletion of chromosomes 3p, 13q and 17p is noted in almost every SCLC, where antioncogene is suspected to be present, and inactivation of antioncogene may play an important role in development of SCLC. Nucleolar size is the important parameter for proliferative potential of SCLC. The larger the nucleoli, the faster is the growth of SCLC. Phenotypes of SCLC in vitro may be altered by change of microenvironment, although it may be due to the selective growth of a certain clone. SCLC and nervous tissue specific membrane antigen is named cluster 1 SCLC antigen, the monoclonal antibody to which will be utilized for immunohistological diagnosis, imaging and treatment of SCLC. Accumulation of basic knowledge is now leading to reconsideration of histological subtyping of SCLC. PMID- 2549879 TI - [Recent trends of chemotherapy in small cell lung cancer: a slow but steady progress]. AB - The author reviewed some distinct findings concerning chemotherapy of small cell lung cancer (SCLC) which had appeared recently in the articles of journals and proceedings. Of new drugs, teniposide and carboplatin have a significant activity against SCLC, however, the usefulness of these drugs must be determined in the comparative fashion with their mother compounds. Cisplatin plus etoposide (PVP), which was demonstrated to have a distinct activity in previously treated SCLC and to be not fully cross-resistant to CAV, has recently been incorporated into the first-line treatment. PVP has been evaluated as either induction chemotherapy or consolidation therapy, as alternating chemotherapy with CAV and as a chemotherapy regimen in the combined modality treatment with chest irradiation. The data obtained provide results comparable or rather superior to those achieved with CAV or other conventional regimens when used as induction chemotherapy, and provide some encouraging results when used as consolidation chemotherapy and in combination with chest irradiation. Four trials have been conducted to test the usefulness of CAV-PVP alternation, however, the results is still controversial. Recently, two institutions reported the results of pilot study of an intensive weekly chemotherapy. Although the complete response rate of the weekly regimens appears superior to that of conventional regimens, the survival benefit is still obscure because of an inadequate follow-up time. The pace of therapeutic advances in SCLC has slowed and appears to reach a plateau, however, recent articles indicate a slow but steady progress in the treatment of SCLC. PMID- 2549880 TI - [The role of radiotherapy in the treatment of small-cell lung cancer]. AB - The role of radiotherapy in the initial management of small-cell lung cancer (SCLC) is a matter of current debate. Unfortunately, compared to other cell types, radiotherapy for SCLC is limited in its use. The value of radiation therapy in palliation is unquestioned. SCLC is quite sensitive to both radiation therapy and chemotherapy. However, in extensive disease radiation therapy is secondary to combination chemotherapy. In limited disease, it is well documented that thoracic irradiation increases locoregional tumor control and improves the complete remission (CR) rate. However, the patients given high-dose thoracic irradiation concurrently with chemotherapy are the possibility of local toxicity, especially, drug induced pneumonitis. Recently, the optimal sequence of radiation therapy and chemotherapy has been discussed. We believe that it is better to start with chemotherapy. With prolongation of survival related to intensive combination chemotherapy, brain metastases become more frequent. Therefore current therapeutic regimens include prophylactic cranial irradiation (PCI) soon after the attainment of CR, but it has not prolonged survival. So there are conflicting reports about neurotoxicity after PCI. It is difficult to reach any firm conclusion in regard to the value of PCI. PMID- 2549881 TI - [Surgical treatment of small cell lung cancer]. AB - Thirty-four small cell lung cancer (SCLC) patients underwent surgery in Kyoto University from 1976 to 1989. The mean age was 62 years males numbered 25 cases, and females 9. All cases were LD except 2 rib-metastasized patients. Seventeen patients were in PS 0, and 17 in PS 1. Twenty-eight patients were treated with chemotherapy. In the remaining 5 patients 2 were treated by only oral 5-FU derivatives, one died upon operation, one was unable to be treated because of advanced age, and the last one refused chemotherapy. Except for the one operational death, 33 cases were able to be evaluated. Out of 28 patients, 16 were treated only by post-operative chemotherapy. As for operational radicality, 21 out of 31 cases were operated radically (68%). The survival rate of these 33 patients reached 78% for 1 year, 37% for and 5 years, with MSI at 26 months. The neoadjuvant (+) group showed a better prognosis than the non-non-neoadjuvant group. PMID- 2549882 TI - [Clinical significance of serum NCC-ST 439 as a tumor marker for colorectal cancer]. AB - Clinical significance of preoperative serum NCC-ST 439 level was studied in 119 cases of colorectal cancer (90 of primary, 29 cases of recurrent). The positive rates for serum NCC-ST 439 were 28.9% in primary and 65.5% in recurrent cases. The false positive rates for benign disease were 5.6%. These rates were low when compared with those for other tumor markers. The positive rates for serum NCC-ST 439 exhibited a strong correlation with wall invasion, lymph node and liver metastases. A combination assay of NCC-ST 439, CEA and CA 19-9 produced a high positive rates as 43.3% in primary and 86.2% in recurrent cases. These results demonstrate that measurement of serum NCC-ST 439 may be useful for cancer staging and improves the diagnostic rate in combination with CEA and CA 19-9. PMID- 2549884 TI - B virus (Herpesvirus simiae) and human infection. AB - B virus (Herpesvirus simiae) infections in macaque colonies are common. Herpetiform lesions as well as asymptomatic shedding of virus in bodily secretions from macaques pose a risk to animal handlers and laboratory workers. Fatal encephalitis in humans infected with B virus has occurred. Dermatologists may become involved in the initial evaluation of animal handlers exposed to this virus through bites or infectious secretions. PMID- 2549883 TI - Unusual cutaneous cytomegalovirus involvement in patients with acquired immunodeficiency syndrome. AB - Two patients with acquired immunodeficiency syndrome presented unusual keratotic cutaneous lesions with a protracted course. Pathologic examination in both patients, cultures, and DNA hybridization techniques of skin biopsy specimens in the second patient were characteristic of cytomegalovirus cutaneous infection. Cytomegalovirus skin lesions are rarely described in acquired immunodeficiency syndrome in contrast with the high frequency of ocular and visceral involvement. Skin biopsy specimens may lead to early diagnosis of cytomegalovirus disseminated disease and to specific treatment. PMID- 2549885 TI - Stimulation of prostaglandin E adenylate cyclase response in pig epidermis by hydrocortisone. PMID- 2549886 TI - Phospholipase C activity in human placental membrane. AB - Receptor-stimulated hydrolysis of polyphosphoinositides by action of phospholipase C appears to be an important mediator of cell activation through the generation of the second messengers, in particular inositol triphosphate (IP3). In order to understand placental function better, activity of IP3 production from membrane in cell-free system was examined. Incubation of membrane preparation from [3H]inositol-labelled human placenta with Ca2+ in the presence of 1 mM ATP and 1 mM GTP resulted in the rapid production of IP3 in a dose dependent manner; half-maximal effect occurred at 10 microM. On the other hand, little effect was observed in the case of membrane prepared from [3H]arachidonic acid-labelled placenta, suggesting higher requirement of Ca2+ for phospholipase A2 activation. These data suggest that placenta contains phospholipase C hydrolyzing polyphosphoinositide at physiological concentration of Ca2. This is the first report to provide direct evidence of transmembrane signalling mechanisms in the human placenta, and may provide a clue to the etiology of placental disorders. PMID- 2549887 TI - Uremic neuropathy and treatment with renal transplantation. AB - Uremic neuropathy is one of the most debilitating symptoms associated with end stage renal disease. Severity can range from mild weakness in the lower extremities to complete physical disability. The only potential cure for uremic neuropathy is renal transplantation. This article will review the literature and describe the clinical features, the etiology, and the treatment of uremic neuropathy. A case history will follow to illustrate the dramatic change in a patient who was referred to a North Carolina institution with severe uremic neuropathy for renal transplantation. PMID- 2549888 TI - Crystal deposition in hypophosphatasia: a reappraisal. AB - Six subjects (three female, three male; age range 38-85 years) with adult onset hypophosphatasia are described. Three presented atypically with calcific periarthritis (due to apatite) in the absence of osteopenia; two had classical presentation with osteopenic fracture; and one was the asymptomatic father of one of the patients with calcific periarthritis. All three subjects over age 70 had isolated polyarticular chondrocalcinosis due to calcium pyrophosphate dihydrate crystal deposition; four of the six had spinal hyperostosis, extensive in two (Forestier's disease). The apparent paradoxical association of hypophosphatasia with calcific periarthritis and spinal hyperostosis is discussed in relation to the known effects of inorganic pyrophosphate on apatite crystal nucleation and growth. PMID- 2549889 TI - Inhibition of neutrophil myeloperoxidase by rabbit anti-(human myeloperoxidase) PMID- 2549890 TI - Ileal pouch-anal anastomosis: comparison of results in familial adenomatous polyposis and chronic ulcerative colitis. AB - The aim of this study was to compare the immediate postoperative results and the long-term outcome of ileal pouch-anal anastomosis in 94 patients with familial adenomatous polyposis to those in 758 patients with ulcerative colitis. Two colitis patients died after operation (0.3%), but no polyposis patients died. Overall operative complications appeared in 26% and 29% of polyposis and colitis patients, respectively (NS). Reoperation for intestinal obstruction did not differ between the two groups, but sepsis requiring reoperation was more common in colitis patients (6%) than in polyposis patients (0%, p less than 0.04). At follow-up (mean, 3 years), polyposis patients had fewer daytime stools (4.5 stools per day), less nighttime fecal spotting (26%), and less pouchitis (7%) than colitis patients (5.8 stools per day; spotting, 40%; pouchitis, 22%; p less than 0.002). The conclusion was that polyposis patients tolerated the operation better and had less long-term disability than did colitis patients. The data suggest that postoperative sepsis, daytime stooling frequency, nocturnal incontinence, and pouchitis may be, at least in part, disease related and not surgeon or operation related. PMID- 2549892 TI - Alterations of cardiac adrenoceptors and calcium channels subsequent to production of aortic insufficiency in rats. AB - Aortic insufficiency (AI) was produced in rats by puncturing one of the aortic valves with a plastic rod inserted from the right carotid artery, and the subsequent changes in alpha- and beta-adrenoceptors and Ca2+ channels were investigated by means of radioligand binding assays. The positive inotropic effects of methoxamine (Met), isoproterenol (Iso) and Ca2+ were also examined in isolated left papillary muscles. The ratio of the whole heart weight to body weight in AI rats increased significantly as compared with sham-operated rats from 1 till 8 weeks after operation, showing that a cardiac hypertrophy had occurred due to the volume overload. Maximum binding sites (Bmax) for 3H-prazosin in cardiac ventricular muscles of AI rats significantly increased at 4 weeks and returned to the control level at 8 weeks when the AI was still patent. Bmax for 3H-dihydroalprenolol (3H-DHA) of AI rats significantly increased at 2 and 4 weeks and returned to the control level at 8 weeks. Bmax for 3H-nitrendipine binding of AI rats significantly decreased at 2 weeks and returned to the control level at 4 weeks. Affinities for these bindings, as expressed by values of dissociation constants, showed no significant changes. The sensitivities of papillary muscles to Met and Iso were significantly increased in AI rats at 4 weeks. Plasma immunoreactive alpha-atrial natriuretic polypeptide (alpha-ANP) concentration of AI rats tended to increase from 1 till 3 weeks and returned to the control level at 4 weeks but these changes were not significant. We conclude that the synthesis of new adrenoceptors keeps pace with the hypertrophic process and may even overshoot during a short period. PMID- 2549891 TI - Sequential patterns of eicosanoid, platelet, and neutrophil interactions in the evolution of the fulminant post-traumatic adult respiratory distress syndrome. AB - Thirty multiply injured blunt-trauma patients at high risk for development of ARDS (multisystem trauma including more than one organ or extremity, Injury Severity Score of 26 or more, hypotension and need for 1500 mL or more blood within the first hour after admission, and PaO2 less than or equal to 70 torr) were studied sequentially with blood and physiologic evaluations beginning immediately after injury and every eight hours for eight days, or until death, to study the evolution of the ARDS process. Mixed venous blood samples were obtained for eicosanoids PGE2, PGF2 alpha, thromboxane B2, PGI2 (6-KetoPGF1 alpha) and leukotriene B4 (LTB4). Platelet (PLAT), and neutrophil (WBC) counts were also done and plasma elastase was measured. At 7:00 AM each day patient neutrophils were obtained for a study of zymosan-activated superoxide production using a chemiluminescence assay. These data were correlated with physiologic measurements of the Respiratory Index (RI), per cent pulmonary shunt (QS/QT), and respiratory compliance measures. Seven patients developed a fulminant post-traumatic ARDS syndrome within 96 hours after injury. Twelve patients without ARDS developed sepsis (TS) four or more days after injury, and 11 had uncomplicated postinjury courses (TR). Compared to both TR and TS, ARDS had a significant (p less than 0.01) rise in neutrophil superoxide production beginning on day 2 through day 4 after injury. This was preceded by rises in PGE2 and LTB4, which were significantly correlated with subsequent falls in PLAT and WBC and rises in TXB2, PGF1, and superoxide production and followed by increases in RI, QS/QT, and a fall in compliance. The significant difference in the pattern and sequence of events in ARDS compared to TR and TS patients suggests that in ARDS the earliest event may be related to peripheral release of PGE2 and LTB4 due to platelet activation and lung sequestration with release of PGF2 alpha, and by aggregation and leukocyte adherence with release of elastase. However, fulminant ARDS mortality appears to be related to the subsequent amplification of the LTB4 leukocyte activation with superoxide production that does not achieve significance before the second day after injury and rises to a maximum by day 4 after injury. These data suggest that post-trauma ARDS follows a different evolutionary pattern than that reported in animal models and is also different from that seen in human TS or TR patients.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2549893 TI - In vitro comparison of the neuromuscular antinicotinic and intestinal antimuscarinic effects of different nondepolarizing muscle relaxants. AB - The postsynaptic antimuscarinic properties of different nondepolarizing muscle relaxants were compared with their postsynaptic antinicotinic effect. d Tubocurarine, pipecuronium and vecuronium were the most selective antagonists on postsynaptic nicotinic receptors. Gallamine, diadonium and Duador (RGH-4201) had relatively greater effect on postsynaptic muscarinic receptors. Therefore, much less side effect is expected to occur when pipecuronium, d-tubocurarine or vecuronium are used. PMID- 2549894 TI - Antiepileptic effects of the excitatory amino acid antagonist cis-2,3 piperidine dicarboxylic acid depend on non-N-methyl-d-aspartate receptors. AB - The effects of the broad spectrum antagonist of the excitatory amino acid neurotransmission, the cis-2,3 piperidine-dicarboxylic acid (cis-2,3 PDA) were investigated on the epileptiform activity induced in vitro by different treatments on rat hippocampal slices. The drug reduced the bursting duration by 45% and the occurrence of the additional population spikes by 50% compared with controls using 1 microM kainic acid to induce epileptiform bursting. At the same concentrations (50-100 microM) cis-2,3 PDA did not significantly affect the magnesium free-induced epileptiform activity. At higher concentrations (200-400 microM), the drug was able to reduce by 25% the bursting duration and the occurrence of the additional population spikes using 1 mM penicillin to induce epileptiform bursting. Our data indicate that the broad spectrum antagonist of the excitatory amino acid transmission, cis-2,3 PDA, presents a low antiepileptic activity that could be related to an influence on "non N-methyl-d-aspartate" (NMDA) receptors. PMID- 2549895 TI - The effect of diazepam on atrial automaticity is not mediated by benzodiazepine receptors. AB - The effect of diazepam on nomotopic cardiac automaticity was examined in the spontaneously beating isolated right atria of the rat. We also investigated whether "central"- or "peripheral"-type benzodiazepine receptors are involved and studied the role of calcium and adenosine in the possible effect of diazepam by the interaction of this drug with either GABA, picrotoxin, RO 15-1788, PK 11195, diltiazem or theophylline. Diazepam (400-500 microM) completely abolished atrial automaticity. This effect was not modified by the presence of either RO 15-1788 (30 microM), PK 11195 (0.1 microM), GABA (100 microM), picrotoxin (7 microM), theophylline (5 microM), diltiazem (0.6 microM) or hypercalcic media (3.6 mM). It is concluded that the diazepam-induced reduction of atrial frequency is mediated neither by "central"- nor "peripheral"- type benzodiazepine receptors, adenosine or calcium and could be due to a nonspecific interaction with the cell membrane. PMID- 2549897 TI - Arterial hypertension as a paraneoplastic phenomenon in hepatocellular carcinoma. AB - Three South African blacks with hepatocellular carcinoma and arterial hypertension are described. Plasma angiotensinogen (renin substrate) concentrations were increased eightfold to 10-fold in the two patients in whom these concentrations were measured. One of these two patients also showed a 34 fold rise in plasma inactive, active, and total renin concentrations, and an elevated plasma renin activity (2.73 ng.L-1.s-1 angiotensin l/mL/h). Inactive renin (prorenin) constituted 90% of the total plasma renin concentration. In the third patient only plasma renin activity was measured, and this was considerably raised (6.05 ng.L-1.s-1; angiotensin l/mL/h). Thus, the arterial hypertension that rarely complicates hepatocellular carcinoma may be caused either by a combination of eutopic synthesis of excessive quantities of angiotensinogen and ectopic production and secretion of active renin by malignant hepatocytes, or by eutopic production of angiotensinogen alone. PMID- 2549896 TI - Gastrointestinal cytomegalovirus infection in heart and heart-lung transplant recipients. AB - Cytomegalovirus (CMV) causes major morbidity in organ transplant recipients. Gastrointestinal disease was the most prominent manifestation of CMV infection in a population of heart and heart-lung transplant patients, with an incidence of 9.9%, compared with pneumonitis (4.0%) and retinitis (0%), and occurred most frequently in CMV-seronegative recipients of organs from CMV-seropositive donors. Clinical manifestations included gastritis (nine patients), gastric ulceration (four patients), duodenitis (three patients), esophagitis (one patient), pyloric perforation (one patient), and colonic hemorrhage (one patient). Patients with gastrointestinal CMV infection were treated with intravenous ganciclovir sodium therapy, 5 mg/kg twice daily, for 2 to 8 weeks, with positive clinical, endoscopic, histologic, and virologic responses. Relapses occurred in four of nine patients who were followed up for a median period of 18 months. Retreatment resulted in healing of endoscopic lesions and in viral clearing. We conclude that early endoscopic evaluation for CMV is indicated in heart and heart-lung transplant patients with gastrointestinal symptoms. This study further suggests that intravenous ganciclovir therapy is effective for the treatment of gastrointestinal CMV in these patients. PMID- 2549898 TI - Injections of beta-adrenergic substances in the locus coeruleus affect the gain of vestibulospinal reflexes in decerebrate cats. AB - 1. The tonic discharge of the noradrenergic locus coeruleus (LC) neurons is dampened by norepinephrine (NE) which acts not only on alpha2-adrenoceptors located on the somatodendritic membrane, through mechanisms of recurrent inhibition, but also on beta-receptors. Experiments were performed to find out whether inactivation of LC neurons by local injection of the beta-adrenergic agonist isoproterenol into the LC complex of one side produced changes in posture as well as in the gain of vestibulospinal reflexes acting on forelimb extensors. 2. In precollicular decerebrate cats the amplitude of modulation and thus the gain of the multiunit EMG responses of the forelimb extensor triceps brachii to animal tilt at 0.15 Hz, +/- 10 degrees, leading to sinusoidal stimulation of labyrinth receptors, were quite small. Microinjection of 0.25 microliter of a solution of the beta-adrenergic agonist isoproterenol at the concentration of 4.5 9.0 microgram/microliter of sterile saline into the LC complex of one side decreased the extensor rigidity in the ipsilateral limbs and to a lesser response gain of the ipsilateral triceps brachii to the same parameters of labyrinth stimulation greatly increased (t-test, P less than 0.001); moreover, a slight but significant increase in phase lead of the responses was observed. These findings appeared within 5-10 min after the injection of isoproterenol, fully developed within 20-30 min and persisted for about 2-3 hours after the injection. 3. The increased gain of the vestibulospinal reflexes acting on the triceps brachii did not depend on the decreased postural activity following injection of the beta adrenergic agonist, since it was still observed if the reduced EMG activity of the extensor muscle following the injection was compensated for by an increased static stretch of the muscle. The positive correlation (t-test, P less than 0.001) between gain of the multiunit EMG response of the triceps brachii to animal tilt and base frequency observed in the control experiment disappeared and was substituted by a slight negative correlation (t-test, P less than 0.05) after injection of isoproterenol into the LC complex, probably due to a more prominent recruitment of motor units for low level of background discharge of the muscle. 4. In addition to the effects which involved the triceps brachii ipsilateral to the side of the injection, a smaller but significant increase in response gain affected the contralateral extensor muscle. This increase in gain was also associated with a slight increase in phase lead of the responses.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2549899 TI - Experience with perindopril in normal volunteers. AB - The new orally active angiotensin converting enzyme (ACE) inhibitor perindopril was evaluated in normotensive men. Doses of 2 to 16 mg were given once a day for up to one week. Single oral doses of perindopril were found to blunt the pressor response to exogenous angiotensin I in a dose-dependent manner. The drug-induced ACE inhibition, as estimated by the measurement of plasma ACE activity, was maximal 4 to 8 hours post drug intake. ACE activity was still importantly reduced 24 hours after dosing. Plasma levels of angiotensin II and aldosterone decreased significantly whereas plasma renin activity and blood angiotensin I levels rose during peak ACE inhibition induced by the 4 and 8 mg doses. However, circulating levels of angiotensin II returned to baseline 24 hours after dosing, both on the first day of treatment and after one week of administration. ACE inhibition with perindopril did not affect blood pressure and heart rate in any consistent manner. There was no evidence for drug accumulation during repeated administration. The novel ACE inhibitor was well tolerated and produced no change in routine laboratory tests. The long-acting ACE inhibitor perindopril appears therefore to be effective when given orally in a dose range of 4 to 16 mg. PMID- 2549900 TI - Characterization and localization of phosphatidylglycerophosphate and phosphatidylserine synthases in Rhodobacter sphaeroides. AB - Catalytic properties and membrane associations of the phosphatidylglycerophosphate (PGP) and phosphatidylserine (PS) synthases of Rhodobacter sphaeroides were examined to further characterize sites of phospholipid biosynthesis. In preparations of cytoplasmic membrane (CM) enriched in these activities, apparent Km values of PGP synthase were 90 microM for sn glycerol-3-phosphate and 60 microM for CDP-diacylglycerol; the apparent Km of PS synthase for L-serine was near 165 microM. Both enzymes required Triton X-100 with optimal PS synthase activity at a detergent/CDP-diacylglycerol (mol/mol) ratio of 7.5:1.0, while for optimal PGP synthase, a range of 10-50:1.0 was observed. Unlike the enzyme in Escherichia coli and several other Gram-negative bacteria, the PS synthase activity had a specific requirement for magnesium and was tightly associated with membranes rather than ribosomes in crude cell extracts. Sedimentation studies suggested that the PGP synthase was distributed uniformly over the CM in both chemoheterotrophically and photoheterotrophically grown cells, while the PS synthase was confined mainly to a vesicular CM fraction. Solubilized PGP synthase activity migrated as a single band with a pI value near 5.5 in a chromato-focusing column and 5.8 on isoelectric focusing; in the latter procedure, the pI was shifted to 5.3 in the presence of CDP diacylglycerol. The PGP synthase activity gave rise to a single polypeptide band in lithium dodecyl sulfate-polyacrylamide gel electrophoresis at 4 degrees C. PMID- 2549902 TI - Inhibin and reproduction. First communication: changes to inhibin level in Barki ewes after artificial infection with bluetongue virus. AB - Bluetongue virus type 16, isolated from sheep in Egypt, was injected to 4 normally cycling Barki ewes and caused high levels of inhibin. This was assayed by a biological method, using suppression of the luteinising hormone (LH) of castrated rats. Albumin (fraction 2) was injected to normally cycling ewes. The sera of injected ewes were investigated 1 day after injection and weekly up to the 4th one injection. There was a gradual decrease of LH (4.8 +/- 0.52 I.U./ml serum) until the minimum level (1.17 +/- 0.25 I.U./ml serum) was reached, in comparison to the control serum LH which was 5.26 +/- 0.52 I.U./ml serum during the dioestrous phase. PMID- 2549901 TI - Cyclic AMP, fructose-2,6-bisphosphate and catabolite inactivation of enzymes in the hydrocarbon-assimilating yeast Candida maltosa. AB - The inactivation of fructose-1,6-bisphosphatase, isocitrate lyase and cytoplasmic malate dehydrogenase in Candida maltosa was found to occur after the addition of glucose to starved cells. The concentration of cyclic AMP and fructose-2,6 bisphosphate increased drastically within 30 s when glucose was added to the intact cells of this yeast. From these results it was concluded that catabolite inactivation, with participation of cyclic AMP and fructose-2,6-bisphosphate, is an important control mechanism of the gluconeogenetic sequence in the n-alkane assimilating yeast Candida maltosa, as described for Saccharomyces cerevisiae. PMID- 2549903 TI - Bovine leucosis virus challenge infection of calves following application of BL-3 cells. AB - 8 calves were vaccinated 3 times with 1.5.10(7) BL-3 cells. 2 and 6 weeks, respectively, after the 3rd vaccination each 4 calves were subjected to a challenge infection with 2.5.10(5) blood lymphocytes of a BLV-infected cow. For control a challenge infection was performed on 3 non-vaccinated calves. The vaccination caused in all calves the appearance of complement-dependent cytotoxic antibodies against BL-3 cells but not of antibodies against BLV-gp51. After the challenge infection there occurred in the sera of all vaccinated calves and of all non-vaccinated control animals antibodies against gp51, and BLV could be demonstrated in the cultivated blood lymphocytes of all animals. Consequently, the application of BL-3 cells had no protective effect against the BLV infection. PMID- 2549904 TI - Determination of avian leucosis virus in chicken pancreas. AB - 30 one-day-old chickens of the commercial HX-SL hybrid from the incubator house at Premyslovice were infected with tissue culture supernatant of clone LSCC-H32. This tissue culture produced the virus of avian lymphoid leucosis of subgroups A and B. After killing at the age of 21-37 weeks, samples of chick pancreas were examined histopathologically and by electron microscopy. Serological examination of serum and feather pulp for presence of avian sarcoma-leucosis gs antigen was simultaneously performed, using the ELISA test. In the pancreas of the experimental chickens both complete and incomplete viral particles of avian leucosis were found. Most frequently, they were localised in the electron-dense secretions of the lumen of pancreatic acini at apical plasma membranes of acinar cells but also in zymogenic granules and in intercellular spaces. The significance of these findings to possible horizontal transmission is discussed. PMID- 2549905 TI - [The reasons for the occurrence of non-specific reactions in the immunodiffusion test for enzootic bovine leukosis]. AB - A batch of enzootic bovine leucosis antigen earmarked for immunodiffusion testing exhibited unspecific responses to cattle sera, which prompted an investigation of the underlying causes. Evidence was produced, for the first time ever, that such reactions were attributable to an antigen-antibody reaction caused by Mycoplasma arginini. PMID- 2549906 TI - The serological response to foot-and-mouth disease vaccination is not affected by simultaneous infectious bovine rhinotracheitis/adenovirus/parainfluenza 3 vaccination. AB - Young calves were simultaneously vaccinated by subcutaneous route against foot and-mouth disease (FMD) and against infectious bovine rhinotracheitis/adenovirus/parainfluenza-3 (IBR/Adeno/PI-3) by intranasal route. The serological response against the 3 FMD virus types of the FMD vaccine was clearly positive. There was no significant difference between results of simultaneous FMD and IBR/Adeno/PI-3 vaccination and FMD vaccination only. PMID- 2549908 TI - Thecal metaplasia in the adrenal gland of a man with acquired bilateral testicular atrophy. AB - Thecal metaplasia of the adrenal was observed as an incidental autopsy finding in a 77-year-old man with acquired bilateral testicular atrophy, probably secondary to previous bilateral inguinal herniorrhaphy. The metaplastic focus was composed of interlacing bundles of spindle cells attached to the adrenal capsule, strongly resembling ovarian theca. A related finding was hyperplasia of the anterior pituitary, involving particularly the basophils. We believe that the acquired gonadal failure of this man may have led to increased activity of the pituitary gonadotropes. Blastema cells outside of the testes with the ability to form gonadal stroma, such as cells of the adrenal capsule, were thereby stimulated to undergo metaplasia, giving rise to ovarian theca. Similar thecal metaplasia has been described previously in the adrenals of postmenopausal women. Whether the metaplastic foci are capable of hormone production is not known. Since this patient had an occult prostatic carcinoma at autopsy, potential hormone production by such metaplastic gonadal stroma is clinically relevant. PMID- 2549907 TI - Pituitary pathology in acquired immunodeficiency syndrome. AB - Pituitary morphology was studied in 49 autopsied patients with acquired immunodeficiency syndrome. Direct infectious involvement was noted in six adenohypophyses (12%), including five cases by cytomegalovirus and one by Pneumocystis carinii. Two cases with neurohypophysial lesions presumably caused by cytomegalovirus and one questionable case of Toxoplasma gondii were also observed. In all instances these changes were associated with generalized and/or cerebral infection by these same agents. Neither Kaposi's sarcoma nor malignant lymphoma was encountered in the pituitary glands. Acute necrotic foci, presumably due to infarction, were noted in four cases. Four pituitary microadenomas (8%) and four hyperplastic nodules were identified. The incidence of such noninfectious lesions, as well as the prevalence and distribution of the various immunoreactive adenohypophysial cell types, were similar to those seen in the pituitary glands of age-matched male control patients. PMID- 2549909 TI - Atypical congenital mesoblastic nephroma. Report of a case with karyotypic and flow cytometric analysis. AB - Atypical congenital mesoblastic nephroma is a rare infantile renal tumor that may behave aggressively in older infants. A case of atypical congenital mesoblastic nephroma occurring in an 8-month-old hispanic male was analyzed by routine histopathologic, cytogenetic, and retrospective flow cytometric analysis for DNA ploidy. Light microscopy revealed marked hypercellularity. The karyotype was abnormal, with the following configuration: 45,XY,-1,-3,-9,-9,-15,-17, 21,+del(1)(q21q25),+der(3), t(3;9;15)(q23;p22;q11),+der(9),t(3;9;15) (q23;p22;q11),+der(9),t(9;?) (p?22;?),+r21, + mar. Retrospective DNA ploidy analysis revealed a DNA index of 1.0. The significance of karyotypic changes occurring in mesenchymal renal tumors of this type is currently unknown. Cytogenetic analysis might be of prognostic value in these potentially aggressive tumors. PMID- 2549910 TI - An unexpected cause of pathologic hip fracture. Malignant fibrous histiocytoma. AB - We describe a case of a 68-year-old patient presenting with pathological hip fracture and multiple pulmonary metastases, who has been operated without a prior histological diagnosis. The hip lesion was thought to be a metastasis of an unknown primary tumor. The proximal part of the femur was resected and replaced by a Muller mega-prosthesis. Histological analysis of the resected bone revealed a malignant fibrous histiocytoma, a rare but very aggressive bone tumor. The patient died three weeks after operation of widespread metastases. By presenting this case report, we want to stress the importance of pretreatment histological diagnosis of osteolytic bone lesions in older patients with metastases. PMID- 2549911 TI - Injection-related ulnar neuropathy. AB - Two previously healthy people sustained mild ulnar mononeuropathies at the level of the upper arm after injections by the same nurse. In both, the nurse was attempting to inject into the middle deltoid; the ulnar nerve was presumably reached by standing at the patient's side and injecting "sidearm" into the upper arm. These cases point out the careful attention to local anatomy required for those administering injections, and the importance of proper positioning. PMID- 2549912 TI - Hepatic resection in 120 patients with hepatocellular carcinoma. AB - During the 11-year period from 1977 through 1987, hepatic resections were carried out in 120 patients with hepatocellular carcinoma (HCC). Twenty-five had HCCs smaller than 5 cm in diameter. There were 97 male and 23 female patients, with an average age of 51.5 years. Among them, 45.8% had liver cirrhosis and 80.8% were positive for hepatitis B surface antigen. Fourteen with ruptured HCCs underwent hepatic resection to control the intra-abdominal hemorrhage. Operative mortality within one month after surgery was 4.1%. The postoperative course was complicated by pleural effusion in 5.8%, subphrenic abscess in 2.5%, postoperative bleeding in 1.6%, hepatic failure in 1.6%, and bile leakage in 0.8% of the patients. The overall five-year survival rate in this series was 25.9%, while survival for the last five years was better (42.3% vs 11.9% for patients treated between 1977 and 1982). The cumulative survival rate had no relation to tumor rupture or liver cirrhosis. The group of patients with smaller tumors (diameter, less than 5 cm) or without vascular invasion by tumor had better survival. PMID- 2549913 TI - Improved early results of elective hepatic resection for liver tumors. One hundred consecutive hepatectomies in cirrhotic and noncirrhotic patients. AB - Liver resection for a neoplasm was performed in 100 patients between 1979 and 1987. There were 43 hepatocellular carcinomas (70% of them arising in patients with cirrhosis), 28 metastases from colorectal cancers, 20 benign tumors, and 9 miscellaneous tumors. Forty-nine patients had a major liver resection, 36 a segmentectomy, and 15 a nonanatomic liver resection. Great care was taken to avoid intraoperative and postoperative bleeding, including late ligation of the hepatic vein, the use of Kelly fracture and resorbable clips for hemostasis of transection planes in 74 patients, and temporary clamping of the portal pedicle in 22. Drainage of the abdomen was avoided in 21 patients to prevent ascitic leakage to decrease the postoperative hospital stay. Operative mortality was 1%. There were eight major complications, including one bile leak, one subphrenic abscess, and three subphrenic hematomas. The use of resorbable clips significantly reduced operative time and transfusion requirements. In patients with cirrhosis, temporary clamping of the hepatic pedicle significantly decreased blood loss. Avoiding drainage significantly decreased the postoperative hospital stay. PMID- 2549915 TI - Sulfhydryl-induced restoration of myocardial contractility after alteration by mercury. PMID- 2549914 TI - Influence of cytochrome P-450 inhibitors on the inhalative uptake of methyl chloride and methylene chloride in male B6C3F1 mice. PMID- 2549916 TI - Stress hormone changes after toxic doses of new anticancer drugs. PMID- 2549917 TI - Lack of stress-induced neuroendocrine changes after prolonged drinking of acid water in the rat. PMID- 2549918 TI - Denaturation of poliovirus procapsids. AB - The denaturation of poliovirus procapsids at pH 6.5 was studied, both in the frozen and liquid condition. Denaturation involved alteration of antigenic and physical features (isoelectric pH, alkali dissociability, sedimentation coefficient). Magnesium was a stabilizing factor, and the sedimentation coefficient was the most denaturation-sensitive feature. PMID- 2549919 TI - Changes in the bovine herpesvirus 1 genome during acute infection, after reactivation from latency, and after superinfection in the host animal. AB - Three subtypes, as defined by HindIII restriction endonuclease (RE) analysis patterns, of bovine herpesvirus 1 (BHV 1) were used to inoculate seronegative, BHV 1-free cattle. These included: infectious bovine rhinotracheitis virus (IBRV), subtype 1.1; infectious pustular vulvovaginitis virus (IPPV) isolate K22, subtype 1.2b; and IPVV isolate FI, subtype 1.2a. Nasal, vaginal, and buffy coat samples were taken for virus isolation from each animal. RE analysis was done on virus isolates collected during acute infection, after reactivation from latency, and after reactivation followed by superinfection with a subtype of BHV 1 that differed from the primary inoculation virus. Changes occurred in the BHV 1 genome after only 1 passage in the host animal, and varied from tissue to tissue within the same animal. Viruses reactivated from latency also displayed genome variability. Only animals that received IPVV as the primary inoculation virus were successfully superinfected. After superinfection, cattle shed both superinfecting and reactivated viruses, and genome variability was observed. These data suggest that the application of RE analysis in diagnostic and epidemiologic studies of BHV 1 is limited to analysis between types and subtypes, and is not applicable for the examination of isolates from within a BHV 1 subtype. PMID- 2549920 TI - Reiterated sequences of herpes simplex virus type 1 (HSV-1) genome can serve as physical markers for the differentiation of HSV-1 strains. AB - The stability of regions containing tandemly reiterated sequences in the S component of the herpes simplex virus type 1 (HSV-1) genome was determined, by comparing restriction fragments of the regions among sets of HSV-1 isolates derived from a single source. The 6 reiterations examined were grouped into three. Reiteration VII (within protein coding regions of genes US10 and US11) and reiteration IV (within introns of genes US1 and US12) were stable between the isolates (group 1). Regions containing one of four other reiterations were detected as a set of ladder-like fragments. Reiteration II (between "a" sequence and IE 175 gene) and reiteration VI (within an intergenic region on the 3' side of the 3' co-terminal family of genes US10, US11, and US12) (group 3) were more unstable than reiteration I (within "a" sequence) and reiteration III (between "a" sequence and IE175 gene) (group 2). The mode of fluctuation of the reiterations observed within a set of HSV-1 strains isolated from an individual was similar to that observed between HSV-1 single-plaque clones separated in cultured cells. These reiterations, except for group 3, can serve as sensitive and convenient markers for differentiating HSV-1 strains. PMID- 2549921 TI - Alcelaphine herpesviruses 1 and 2 SDS-PAGE analysis of virion polypeptides, restriction endonuclease analysis of genomic DNA and virus replication restriction in different cell types. AB - Herpesviruses have been isolated from white-tailed, white-bearded and blue wildebeest, as well as from Jimela topi and Cape hartebeest. These animals are members of the sub-family Alcelaphinae of the family Bovidae. Viruses isolated from wildebeest cause malignant catarrhal fever (MCF) in susceptible ruminant species. Alcelaphine herpesviruses (AHV) isolated from wildebeest replicate in both fetal aoudad sheep kidney (FAK) cells and bovine embryonic lung (BEL) cells. However, virus isolates from topi and hartebeest, which have not been linked to clinical MCF, replicate only in FAK cells. Buoyant density analysis by analytical ultracentrifugation, restriction endonuclease analysis and blot hybridization of virus genomic DNA from both alcelaphine herpesviruses as well as from bovine herpesviruses 1, 2, and 4 demonstrate that there are two types of alcelaphine herpesviruses, each distinct and different from the other bovine herpesviruses. Genomic size of both alcelaphine herpesviruses, estimated from DNA restriction fragments, is approximately 110 kilobase pairs. Alcelaphine herpesvirus DNA resembles Herpesvirus saimiri DNA during equilibrium sedimentation in that the majority of the DNA bands as a light (L) fraction with a minor heavy (H) component. Polyacrylamide gel analysis of virion proteins indicates that both viruses have distinct patterns, each consisting of 36 polypeptides ranging in molecular weight from 12,000 to 275,000. Virus isolates from wildebeest have been designated AHV-1, while viruses isolated from topi and hartebeest have been designated AHV-2. PMID- 2549922 TI - Hemagglutination with pseudorabies virus. Brief report. AB - Pseudorabies virus grown in CPK cell cultures was tested for hemagglutination (HA) with erythrocytes of a variety of species at 4 degrees C, 25 degrees C and 37 degrees C. HA was observed at all temperatures with mouse erythrocytes but not with cattle, sheep, goat, swine, cat, rabbit, guinea pig, rat, mongolian gerbil, chicken, and goose erythrocytes. Mice showed a strain variation in agglutinability of their erythrocytes, requiring selection of mice to obtain erythrocytes for HA. The HA reaction was inhibited by specific antiserum. Some factors involved in the HA and HA-inhibition (HI) were investigated and standard HA and HI tests were established. HI antibody titers of individual pig sera showed a significant positive correlation with their neutralizing antibody titers. PMID- 2549923 TI - [Morphology of borderline mucinous ovarian tumors]. AB - The examination of structures of the tumor tissues taken from 47 patients with borderline mucinous tumours of the ovary showed that the tumors belonging to this group, like their serous analogues, were an indistinctly limited nosological entity that characterizes a particular stage in the malignant transformation of the epithelium as evidenced by a wide range of structures that are interchangable from mucinous adenofibroma to mucinous carcinoma. Death may result from pseudomyxoma or dissemination that follows the pattern seen in cancer. The latter generally stems from small-sized areas of occult cancer which are detectable from a retrospective analysis of the material available. Intraoperative laceration of the tumor makes no contribution to the occurrence of peritoneal pseudomyxoma. The results of the histochemical study into the mucosa indicated that the tumor cells were heterogeneous in mucosaccharides. The cells which were completely free from acid fractions in the presence of neutral mucins were identified. Disregarding the features indicated, one can reasonably reject the fact that the neoplasm is of mucinous character as judged from alcian blue-stained specimens. PMID- 2549924 TI - Toxic and pet exposures in amyotrophic lateral sclerosis. PMID- 2549925 TI - Corticotropin is superior to corticosteroids in the treatment of MS. PMID- 2549927 TI - Long-term effects of megavoltage radiotherapy in acromegaly. AB - The progress of 41 Chinese patients with acromegaly treated with megavoltage radiotherapy was reviewed after a mean follow-up of 4.5 (one-ten) years. Nine received prior surgery. Radiotherapy was delivered by a 3-field technique to a total of 4000-5000 cGy in 25 fractions. By life table analysis successful treatment of growth hormone (GH) hypersecretion, as defined by a mean GH concentration of less than or equal to 10 mu/L, could be expected in 6, 11, 26, 64 and 67% of the patients after one, two, five, eight and ten years respectively. Median intervals before achieving a mean GH level of less than or equal to 10 mu/L were 6.6 and 8.6 years following radiotherapy with the higher (4500-5000 cGy) and lower (4000 cGy) doses respectively, suggesting a tendency towards earlier response following radiotherapy with the higher dose. The prevalence of acquired hypopituitarism in patients followed up for over five years was 40% for gonadotrophins, 30% for TSH and 20% for ACTH deficiency respectively. In the majority of patients, acquired hypopituitarism was not apparent within five years after radiotherapy. No mortality or major side effects were noted following radiotherapy. In 34 patients on long-term bromocriptine treatment, mean GH concentrations were normalised in 26.5% of patients. We conclude that with judicious selection of patients and use of adjunctive medical therapy, megavoltage radiotherapy remains a safe and satisfactory form of treatment for acromegaly especially if expert transphenoidal surgery is not readily available. PMID- 2549926 TI - Ouabain binding in the human brain. Effects of Alzheimer's disease and aging. AB - We studied Na+,K+-adenosine triphosphatase by assaying specific tritiated ouabain binding in the frontal cortex, temporal cortex, hippocampus, putamen, cerebellum, and cerebral microvessels in subjects with Alzheimer's disease and control subjects. Ouabain binds specifically, in a saturable manner, and with a high affinity to a single class of binding sites in all the tissues studied. The density of ouabain binding sites was highest in cerebellum and frontal cortex (approximately 40 pmol/mg of protein); intermediate in temporal cortex, hippocampus, and putamen; and lowest in brain microvessels (approximately 8 pmol/mg of protein). The dissociation constant of binding was about 30 nmol/L in all tissues. In control subjects, there were no age-related alterations in ouabain binding, nor was there any correlation between ouabain binding and postmortem delay. However, there was a marked decrease in brain ouabain binding in subjects with Alzheimer's disease when compared with age-matched controls, especially in the cerebral cortex. Ouabain binding was also significantly decreased in the cerebellum and putamen of subjects with Alzheimer's disease even though these brain regions are not particularly affected in this disease. Ouabain binding to brain microvessels, which constitute the blood-brain barrier, was not significantly decreased in subjects with Alzheimer's disease. The decreased specific ouabain binding in the brain of subjects with Alzheimer's disease probably reflects the loss of neuronal membranes. PMID- 2549928 TI - A farming systems study of abortion in dairy cattle on the Atherton Tableland. 3. Metabolic factors. AB - An association with a rank correlation of 0.55 (p = 0.06) was obtained between monthly rainfall and monthly abortions for a group of 24 farms in tropical Australia where cows grazed predominantly summer rain-dependent pasture. A similar association was found among cows grazing nitrogen-fertilised, irrigated pasture in winter in the same area. On farms with irrigated pasture the first service conception and abortion rates in herds which grazed intensively during the night (abortion 8.9%; first service conception 42.9%) differed significantly from those which grazed during the day (abortion 6.4%; first service conception 52.9%), and this suggested that increased abortion rates and decreased first conception rates may be part of the one syndrome. An hypothesis was developed that the feeding of immature nitrogen-fertilised pasture, rich in protein and stimulated by rain or irrigation, is associated with an increased abortion rate and a depressed conception rate. Other evidence included an increased susceptibility of first lactation cows to abortion, a lower abortion rate in late pregnant dry cows fed more mature pasture, a 20% repeat abortion rate in the population sampled and the lack of conclusive evidence for infectious and selected non-infectious agents. There were no other observed clinical signs in most aborting cows. PMID- 2549929 TI - A farming systems study of abortion in dairy cattle on the Atherton Tableland. 4. Pasture composition and plasma progesterone concentrations of pregnant cows in affected herds. AB - To provide more evidence for an apparent association between immature, high protein pasture diets and abortion in dairy cows on the Atherton Tableland in tropical north Queensland, pastures and cows on 4 farms were sampled on 5 occasions during a 7-month period. Pasture samples were analysed for dry matter (DM), protein, nitrate and fibre content. Plasma progesterone concentrations were determined in 18 to 20 pregnant cows. Fifty-four pasture samples (38 grass and 16 legume) were analysed and the distribution for 3 DM ranges was: 10% to 14.9% - 20 samples, 15% to 19.9% - 27 samples and 20% to 24.9% - 7 samples, while the distribution for 4 protein ranges was: 15% to 19% - 5 samples, 20% to 24% - 20 samples, 25% to 29% - 21 samples and 30% to 34% - 8 samples. Both pasture protein and DM content fluctuated with time. Pasture nitrate was low and insignificant while fibre (cell wall) levels appeared to be satisfactory for ruminant nutrition. Significant differences occurred between the mean plasma progesterone levels on 3 of the 4 farms. Raised levels in pregnant cows suggested the possibility of increased progesterone production in response to a previous low progesterone crisis or, alternatively, a decreased metabolic clearance rate of progesterone in pregnant cows on a declining plane of nutrition towards the end of the summer wet season. Progesterone data were not available from cows which previously aborted.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2549930 TI - Evidence for Australia-wide canine parainfluenza infection. PMID- 2549931 TI - Diagnosis of a field outbreak of avian encephalomyelitis by enzyme-linked immunosorbent assay. PMID- 2549932 TI - A survey of skin neoplasms in dogs from the Brisbane region. PMID- 2549933 TI - Cell lines for growth of sheep viruses. PMID- 2549934 TI - Comparative microscopic lesions in reoviral and staphylococcal tenosynovitis. AB - Experimental inoculation of 1-day-old male broiler chickens with avian reo-virus or Staphylococcus aureus caused tenosynovitis of the gastrocnemius and digital flexor tendons. Reovirus inoculation by either the oral or footpad route initiated a diffuse lymphocytic infiltration in the peritendineum, synovial membrane, and epitenon from 1 to 5 weeks postinoculation (PI). Heterophils were not a predominant feature of the inflammatory response, but when present they were localized with fibrin in and around synovial spaces. The prevalence of microscopic tendon lesions was less common with staphylococcal infection than with reovirus infection. With staphylococcus, lesions were localized to the synovial space and membranes and were characterized by heterophils and fibrin but few lymphocytes. Synovial cell hyperplasia and bursal atrophy were common in both groups. From 10 to 20 weeks PI, both groups developed progressive tendon fibrosis. These results indicate that tenosynovitis due to inoculation with reovirus or staphylococcus may be differentiated histologically from 1 through 5 weeks PI. After 10 weeks, this may not be possible, because diffuse fibrosis was the major lesion with both. Perhaps fibrosis predisposes older, heavier broilers to tendon failure and rupture. PMID- 2549935 TI - Infectious anemia caused by a parvovirus-like virus in Georgia broilers. AB - Pale chicks with necrotic dermatitis, small bursas of Fabricius (BFs), small thymuses, pale bone marrow, and watery blood were suspected of having parvovirus like virus- (PVLV) associated disease. Histologic lesions included atrophy or hypoplasia of thymuses and BFs, and septic necrotizing clostridial dermatitis and hepatitis. Clostridium perfringens was cultured from skin and liver. A PVLV was isolated in a Marek's disease tumor cell line (MDCC-MSB1) culture and was identified by physicochemical, immunofluorescent, and morphologic features. This isolate was named GA-1 PVLV. Specific-antibody-negative chicks and embryos infected with heat- or chloroform-treated GA-1 PVLV developed anemia at the same rate. Control chicks never were anemic. This is the first isolation of PVLV from clinically ill chickens in the United States and the first report of PVLV-induced anemia in chickens in the Western Hemisphere. PMID- 2549936 TI - Observations of experimentally induced quail bronchitis. AB - Quail bronchitis was experimentally reproduced in captive-reared bobwhite quails (Colinus virginianus). Quails were inoculated with 10(6) mean tissue culture infective doses of quail bronchitis virus at 1,3,6, or 9 weeks of age by the intratracheal, intraperitoneal, or subcutaneous route. Clinical signs were minimal, but occasionally birds were ruffled, exhibited open-mouthed breathing, and developed "snicks." Mortality rates of quails inoculated at 1 or 3 weeks ranged from 7% to 87%. Quails inoculated at 6 or 9 weeks of age had mortality rates from 0% to 20%. Mean body weights of survivors that had been inoculated at 3 or 6 weeks were significantly less than those of controls (P less than 0.05). No significant differences in body weight were detected between quails inoculated at 1 or 9 weeks and their uninoculated controls. Antibodies to group I adenovirus were detected by agar gel precipitation in 87.5% of birds that survived infection. PMID- 2549937 TI - Structural proteins of two different plaque-size phenotypes of fowlpox virus. AB - Structural polypeptides of two plaque-purified variant isolates of fowlpox virus differing in plaque morphology and size were examined by Coomassie blue-staining and immunoblot analysis of purified virions. A total of 30 structural polypeptides were observed, ranging in molecular weight from 14,100 to 122,600. A late polypeptide of 36,400 molecular weight was quite prominent in the small plaque clone but absent in the large-plaque clone. Two other polypeptides, of 33,700 and 34,800 molecular weight, were present in virions from large-plaque virus and cell lysates of both clones but were absent in the small-plaque virions. These differences were observed whether the viruses were grown in chorioallantoic membrane or in chicken embryo fibroblast cultures. No difference was observed between the growth curves of the two virus clones. Differences observed in the polypeptides of the two viruses may be due to changes in the less conserved regions of viral DNA and may be used for differentiation of virus isolates. PMID- 2549938 TI - In vivo studies with dimethyldithiocarbamate, a possible new antimicrobial for use against Aspergillus fumigatus in poultry. AB - Dimethyldithiocarbamate (DmDTC), the carbamate analogue, was tested for therapeutic efficacy in a series of in vivo challenge trials using 5- and 10-week old white leghorn chickens. Challenge organisms were Pasteurella multocida X-73, Escherichia coli O1:K1, and Aspergillus fumigatus. Birds were evaluated for survival rates, lesion scores, and the rate at which the bacteria or mold could be reisolated following challenge. Results showed DmDTC to be ineffective against P. multocida and E. coli at the treatment levels and in the form used in these trials, but DmDTC significantly reduced lesion scores and inhibited the rate of isolation of A. fumigatus compared with untreated infected birds. PMID- 2549940 TI - Studies of infectious laryngotracheitis vaccines: immunity in layers. AB - Ten-week-old layer chickens obtained from a commercial source were eye-drop vaccinated with chicken-embryo-origin (CEO) or tissue-culture-origin (TCO) vaccines for infectious laryngotracheitis (ILT). Controls were not vaccinated. Approximately one-third of the layers were challenged with virulent ILT virus at 21, 40, or 60 weeks of age. Serum samples taken from the layers before challenge were used in a virus neutralization (VN) test to determine vaccination titers at those three ages. Both vaccines induced low VN titers (geometric mean titer [GMT] less than 6). At 21 weeks of age, the titers produced by the two vaccines were not significantly different, but at 40 and 60 weeks of age the VN GMT of the CEO vaccinated group was significantly greater than that of the TCO-vaccinated group. The VN GMTs did not drop over time in either group and actually rose between 21 and 60 weeks of age in the CEO group. Both vaccines protected layers against severe challenge with virulent ILT virus, neither being significantly better than the other under these experimental conditions. Unvaccinated sentinel chickens were maintained in contact with the vaccinated layers during three intervals between 1 day and 6 weeks post-vaccination. Diagnostic tests performed on the sentinels to detect lateral spread of vaccine virus from vaccinated to unvaccinated chickens showed scattered positive results. PMID- 2549939 TI - Studies of infectious laryngotracheitis vaccines: immunity in broilers. AB - Broiler chickens were vaccinated at 18 days of age against infectious laryngotracheitis (ILT) using chicken-embryo-origin (CEO) and tissue-culture origin (TCO) vaccines, each vaccine given either by drinking water, spray, or eyedrop. Controls were not vaccinated. The broilers were challenged 3 weeks later with virulent ILT virus (USDA challenge strain). Serum samples taken before challenge were analyzed by a virus neutralization (VN) test to determine titers due to vaccination. Both vaccines, regardless of route of administration, produced low VN titers, geometric mean titer (GMT) being less than 4.0 in all vaccinated groups. When administered by the same route, the CEO vaccine produced higher titers than the TCO vaccine. Titers following drinking-water or eyedrop administration of vaccines were higher than titers following spray vaccination. There was an inverse relationship between pre-challenge VN titers of groups of birds and the percentage of birds in the groups dying from ILT virus challenge. The drinking-water route of vaccination provided the most protection, while the spray provided the least. PMID- 2549942 TI - In vitro and in vivo characterization of avian reoviruses. II. Clinical evaluation of chickens infected with two avian reovirus pathotypes. AB - The effect of two avian reovirus isolates (2408 and 1733) on digestion and nutrient metabolism in infected chickens was assessed by an in vitro absorption assay and clinical blood chemistry analysis. Birds of various ages were inoculated orally and intratracheally with reovirus and sampled periodically for the respective assays. Transitory malabsorption was observed in the duodenum of birds infected with reovirus 2408. Conversely, increased absorption was detected in the ileum of these same birds. Clinical blood chemistry analyses of birds infected with both isolates revealed that severely affected birds had abnormally elevated plasma total protein, plasma albumin, and calcium levels. Decreases were found in percent bone ash and, due to abnormally high globulin levels, in albumin:globulin (A:G) ratios. A significant (P less than 0.05) correlation between body weights and total protein, albumin, A:G ratio, and bone ash was found in infected birds. The most pronounced metabolic and physiologic changes occurred in the severely affected birds, and, in general, pathogenicity of the isolates was reflected by the degree of metabolic change. PMID- 2549941 TI - In vitro and in vivo characterization of avian reoviruses. I. Pathogenicity and antigenic relatedness of several avian reovirus isolates. AB - Pathogenicity, pathogenesis, and antigenic relatedness of four avian reovirus isolates obtained from commercially reared broilers were investigated. Chickens of various ages were inoculated both orally and intratracheally with reovirus. Based on disease signs, mortality, weight depression, tissue lesions, invasiveness, and viral persistence in chickens inoculated at 1 day of age, the isolates were classified as being of low, intermediate, or high pathogenicity. The low-pathogenicity isolate (2177) did not cause mortality, weight depression, or clinical disease. The isolate of intermediate pathogenicity (2035) produced low mortality rates (8%), some weight reduction by 7 weeks postinoculation, and microscopic lesions in the intestine and gastrocnemius tendons. The pathogenic isolates, 2408 and 1733, caused severe clinical disease characterized by stunting, feathering abnormalities, mortality as high as 84%, and microscopic lesions in the liver, intestine, pancreas, and/or gastrocnemius tendon. Highly pathogenic isolates also persisted longer in tissues of infected birds and elicited a more prompt and prolonged antibody response. Birds inoculated at 1 day or 1 week of age were more susceptible to reovirus-induced disease than birds inoculated at 2 weeks, suggesting an age-associated resistance. All isolates produced mortality with equal frequency in embryos. The isolates characterized were found to be antigenically similar based on cross-neutralization and cross protection studies. PMID- 2549943 TI - In vitro and in vivo characterization of avian reoviruses. III. Host factors affecting virulence and persistence. AB - Three avian reovirus isolates (2177, 2035, and 1733) were used to determine the effect of the age of chickens at inoculation on virus virulence and persistence. Groups of specific-pathogen-free leghorns were inoculated with three different reovirus isolates of different levels of pathogenicity at 1 day, 1 week, 2 weeks, 3 weeks, or 4 weeks of age. Tissues were examined for the presence of virus and lesions at regular intervals until 8 weeks postinoculation (PI) and then again at 22 weeks PI. Isolate 1733, which is highly pathogenic, was reisolated from the thymus, trachea, liver, intestine, cecal tonsils, bursa of Fabricius, gastrocnemius tendon, and white blood cells. Microscopic lesions were observed in some tissues, including the thymus, liver, spleen, bursa of Fabricius, and gastrocnemius tendons, when sampled within a 7-day period following inoculation. This isolate persisted and produced microscopic lesions in the gastrocnemius tendons for as long as 22 weeks PI. The isolates of intermediate pathogenicity (2035) or low pathogenicity (2177) were isolated less frequently and from fewer tissues than isolate 1733. Isolate 2035 could be found in the gastrocnemius tendons as long as 7 weeks PI, whereas isolate 2177 was never isolated from the tendons, nor did it produce any notable gross or microscopic tissue changes. Birds inoculated at age 1 week or older with any of the three reovirus pathotypes were more resistant to infection than 1-day-old inoculates, as evidenced by a decrease in virus reisolations and a concurrent reduction in the severity of lesions in selected tissues. PMID- 2549944 TI - Propagation of avian rotavirus in primary chick kidney cell and MA104 cell cultures. AB - The Ch2 strain of avian rotavirus was propagated in primary chick kidney cell (CKC) and MA104 cell cultures in the presence of trypsin. Cultures were evaluated for the presence of rotavirus by an indirect fluorescent antibody (IFA) assay and by an indirect enzyme-linked immunosorbent assay (ELISA). After two passages, the viral titer was significantly higher in CKC than MA104 cell cultures. Also, the IFA assay was more sensitive than the indirect ELISA for detecting rotavirus positive cultures. PMID- 2549945 TI - Adenoviral pancreatitis in guinea fowl (Numida meleagris). AB - Necrotizing pancreatitis was observed in 2-week-old Guinea fowl submitted for necropsy and histopathology. Intranuclear inclusion bodies seen histologically in acinar epithelium were examined by electron microscopy and found to contain viruses resembling adenovirus. Adenovirus was isolated in embryonated eggs from the pancreata of affected birds. The adenovirus isolated was not neutralized by chicken antisera developed against 10 known serotypes of group 1 avian adenoviruses. PMID- 2549946 TI - Cholangiocarcinoma in a 4-month-old double yellow-cheeked Amazon parrot (Amazona autumnalis). AB - A cholangiocarcinoma was diagnosed in a 4-month-old double yellow-cheeked Amazon parrot (Amazona autumnalis). Histologically, neoplastic cells effaced hepatocellular architecture. Ultrastructurally, variably dilated bile ductules were lined by simple cuboidal to low columnar ciliated epithelial cells with sparse cellular organelles and prominent junctional complexes. Elongated single cilia arising from basal bodies in the apical cytoplasm were occasionally evident. The presence of such a neoplasm in a 4-month-old parrot is extremely unusual. PMID- 2549947 TI - Low plasma beta-endorphin in post-traumatic stress disorder. AB - We compared serum cortisol, ACTH and plasma beta-endorphin in 21 Post-Traumatic Stress Disorder patients and 20 controls. Although we found no important disturbance in diurnal rhythms, the PTSD patients had significantly higher A.M. serum cortisols compared with controls. Both A.M. and P.M. plasma beta-endorphins in PTSD patients were significantly lower compared with controls. These data suggest that plasma beta-endorphin may be a marker for PTSD and that chronic endogenous opioid depletion may play a role in the pathogenesis and perpetuation of this disorder. PMID- 2549949 TI - Scrape-loaded p21ras down-regulates agonist-stimulated inositol phosphate production by a mechanism involving protein kinase C. AB - The effect of scrape-loaded [Val-12]p21ras on agonist-stimulated phosphatidylinositol 4,5-bisphosphate (PIP2) turnover in Swiss-3T3 cells was studied. Previously [Morris, Price, Lloyd, Marshall & Hall (1989) Oncogene 4, 27 31] we demonstrated that [Val-12]p21ras activates protein kinase C within 10 min of scrape loading. Here, we show that [Val-12]p21ras inhibits bombesin and platelet-derived growth factor-stimulated PIP2 breakdown 1.5-4 h after scrape loading. This effect persisted for at least 18 h and could be mimicked in control cells by activation of protein kinase C with 12-O-tetradecanoyl 13-acetate (TPA) 15 min prior to ligand stimulation. When protein kinase C was down-regulated by chronic TPA treatment, [Val-12]p21ras was no longer able to inhibit agonist stimulated inositol phosphate production. These results indicate that changes in inositol phosphate levels caused by ras protein are probably due to activation of protein kinase C and not to an interaction of ras with phospholipase C. PMID- 2549948 TI - Mammalian protein methylesterase. Physical and enzymic properties. AB - Protein methylesterase (PME) amino acid composition and substrate specificity towards methylated normal and deamidated protein substrates were investigated. The enzyme contained 23% acidic and 5% basic residues. These values are consistent with a pI of 4.45. The product formed from methylated protein by PME was confirmed as methanol by h.p.l.c. The kcat. and Km values for several methylated protein substrates ranged from 20 x 10(-6) to 560 x 10(-6) s-1 and from 0.5 to 64 microM respectively. However, the kcat./Km ratios ranged within one order of magnitude from 11 to 52 M-1.s-1. Results with the irreversible cysteine-proteinase inhibitor E-64 suggested that these low values were in part due to the fact that only one out of 25 molecules in the PME preparations was enzymically active. When PME was incubated with methylated normal and deamidated calmodulin, the enzyme hydrolysed the latter substrate at a higher rate. The Km and kcat. for methylated normal calmodulin were 0.9 microM and 31 x 10(-6) s-1, whereas for methylated deamidated calmodulin values of 1.6 microM and 188 x 10( 6) s-1 were obtained. The kcat./Km ratios for methylated normal and deamidated calmodulin were 34 and 118 M-1.s-1 respectively. By contrast, results with methylated adrenocorticotropic hormone (ACTH) substrates indicated that the main difference between native and deamidated substrates resides in the Km rather than the kcat. The Km for methylated deamidated ACTH was 5-fold lower than that for methylated native ACTH. The kcat./Km ratios for methylated normal and deamidated ACTH were 43 and 185 M-1.s-1 respectively. These results indicate that PME recognizes native and deamidated methylated substrates as two different entities. This suggests that the methyl groups on native calmodulin and ACTH substrates may not be on the same amino acid residues as those on deamidated calmodulin and ACTH substrates. PMID- 2549950 TI - Benzylhydrazine as a pseudo-substrate of bovine serum amine oxidase. AB - Bovine serum amine oxidase is inhibited by benzylhydrazine (BHy), but recovers full activity after a few hours incubation [Hucko-Haas & Reed (1970) Biochem. Biophys. Res. Commun. 38, 396-400]. The first phase of the process, requiring about 15 min, was found to consist of a mechanism-based hydrazine-transfer reaction leading to formation of the hydrazine-bound enzyme, benzaldehyde and H2O2. At variance with the enzymic process, the reaction with O2 preceded the benzaldehyde release. Two reaction intermediates could be characterized by optical spectroscopy and were assigned as the azo derivative and the benzaldehyde hydrazone, the latter one probably being involved in the reaction with O2. No reduction of Cu was detected at any stage. The hydrazine adduct could also be obtained by stoichiometric reaction of hydrazine with the native enzyme. The decay of this species occurred in about 8 h and was not studied in detail. The Cu binding inhibitor NN-diethyldithiocarbamate affected the BHy reaction by stabilizing the benzaldehyde hydrazone form as against the azo derivative and the reaction with O2. However, under these same conditions the initial spectroscopic properties of the diethyldithiocarbamate adduct were recovered if the oxidase was left overnight. The reaction with O2 was abolished only upon removal of at least one Cu atom from the enzyme. On the basis of the failure to detect any change of Cu redox state and the enzyme behaviour in the presence of inhibitors, a reaction mechanism involving the formation of a hydroperoxy intermediate, as in the FAD containing enzymes, is tentatively proposed. PMID- 2549951 TI - Synthesis of a truncated Mr 46,000 mannose 6-phosphate receptor that is secreted and retains ligand binding. AB - The Mr 46,000 mannose 6-phosphate receptor is an integral membrane protein with its ligand-binding site in the ectoplasmic domain. By site-directed mutagenesis, a stop codon was introduced in the receptor cDNA at the border between the ectoplasmic and membrane-spanning domain. The truncated receptor was expressed in three different systems, Xenopus oocytes, COS cells and BHK-21 cells. In all three systems the truncated receptor behaved as a soluble protein. In oocytes only small amounts of the truncated receptor were secreted within 48 h after synthesis. Accumulation of endoglucosaminidase H-sensitive forms of the truncated receptor in oocytes suggested that exit from the endoplasmic reticulum was slowed down. In COS and BHK-21 cells, the truncated receptor was secreted and, as for wild-type receptor, most of the N-linked oligosaccharides were processed to complex forms. Both the intracellularly-retained (oocytes) and the secreted (COS and BHK-21 cells) truncated receptors bound to phosphomannan-Sepharose in a mannose-6-phosphate-dependent manner. Using chemical cross-linking, the truncated receptor was shown to be secreted as a homodimer. PMID- 2549952 TI - DNA sequences of a bovine gene and of two related pseudogenes for the proteolipid subunit of mitochondrial ATP synthase. AB - The dicyclohexylcarbodi-imide-reactive proteolipid is a membrane subunit of mitochondrial ATP synthase. In cows it is encoded by two different nuclear genes known as P1 and P2. These genes are expressed in a tissue-specific fashion which reflects the embryonic origin of the tissues. The proteins that they encode are synthesized in the cytosol, and are precursors of the proteolipid that have different mitochondrial import sequences of 61 and 68 amino acids respectively. By use of gene-specific probes derived from the bovine P2 cDNA, regions containing corresponding parts of the bovine P2 gene have been isolated from a bovine genomic library, and their DNA sequences and those of flanking and intervening regions have been determined. The sequence contains four exons, which represent the cDNA sequence, spread over 3.8 kb of the bovine genome. Two of the introns are in the DNA sequence coding for the mitochondrial import sequence, and a third intron is in a sequence encoding an extramembranous structure between the two putative transmembrane alpha-helical domains of the mature proteolipid. An Alu-type repetitive element was detected at the extreme 5' end of the sequence. The bovine P1 and P2 genes for the dicyclohexylcarbodimide-reactive proteolipid of ATP synthase are members of a multiple gene family that also contains many pseudogenes. The bovine P1 gene has not been isolated, but two distinct P1 pseudogenes have been cloned and their DNA sequences have been determined. Both of them contain 'in-phase' stop codons and frame-shift mutations, and one of them bears the hallmarks of retroposition; it has no introns, it contains a poly(A) tract at its 3' end and it is flanked by direct DNA sequence repeats. The second P1 pseudogene is very unusual. It appears to be derived from a partially processed transcript and contains an intervening DNA sequence of 861 bp that corresponds in position with an intron in the human P1 gene. This pseudogene also could have been introduced by retroposition since its sequence is flanked by short direct repeats. However, it does not contain a poly(A) tract at its 3' end. An alternative, but less likely, explanation is that rather than being a retroposon, this sequence arose by duplication of an expressed gene at a time when it had only one intron. PMID- 2549953 TI - A continuous fluorimetric assay for clostridial collagenase and Pz-peptidase activity. AB - The peptide derivative N alpha-(2,4-dinitrophenyl)-L-prolyl-L-leucyl-glycyl-L prolyl-L-tryptophanyl-D- lysine (Dnp-Pro-Leu-Gly-Pro-Trp-D-Lys) has been found to be a convenient substrate for the assay of clostridial collagenase and Pz peptidase. The substrate shows a 25-fold enhancement of fluorescence (gamma ex. 283 nm, lambda em. 350 nm) following hydrolysis of the Leu2-Gly3 peptide bond. The value of Km for clostridial collagenase was 17 microM. The substrate for the first time makes possible continuous fluorimetric assays for Pz-peptidase and clostridial collagenase. PMID- 2549954 TI - Activation and phosphorylation of the 'dense-vesicle' high-affinity cyclic AMP phosphodiesterase by cyclic AMP-dependent protein kinase. AB - Incubation of a hepatocyte particulate fraction with ATP and the isolated catalytic unit of cyclic AMP-dependent protein kinase (A-kinase) selectively activated the high-affinity 'dense-vesicle' cycle AMP phosphodiesterase. Such activation only occurred if the membranes had been pre-treated with Mg2+. Mg2+ pre-treatment appeared to function by stimulating endogenous phosphatases and did not affect phosphodiesterase activity. Using the antiserum DV4, which specifically immunoprecipitated the 51 and 57 kDa components of the 'dense vesicle' phosphodiesterase from a detergent-solubilized membrane extract, we isolated a 32P-labelled phosphoprotein from 32P-labelled hepatocytes. MgCl2 treatment of such labelled membranes removed 32P from the immunoprecipitated protein. Incubation of the Mg2+-pre-treated membranes with [32P]ATP and A-kinase led to the time-dependent incorporation of label into the 'dense-vesicle' phosphodiesterase, as detected by specific immunoprecipitation with the antiserum DV4. The time-dependences of phosphodiesterase activation and incorporation of label were similar. It is suggested (i) that phosphorylation of the 'dense vesicle' phosphodiesterase by A-kinase leads to its activation, and that such a process accounts for the ability of glucagon and other hormones, which increase intracellular cyclic AMP concentrations, to activate this enzyme, and (ii) that an as yet unidentified kinase can phosphorylate this enzyme without causing any significant change in enzyme activity but which prevents activation and phosphorylation of the phosphodiesterase by A-kinase. PMID- 2549955 TI - Prostaglandin E2 inhibits phosphoinositide metabolism in isolated pancreatic islets. AB - Isolated islets of the rat labelled with myo-[3H]inositol showed decreased accumulation of total inositol phosphates (InsPs) and [3H]polyphosphoinositide hydrolysis in response to glucose after preincubation with prostaglandin E2 (PGE2). The response was concentration-dependent and specific for PGE2. PGE2 did not affect basal [3H]phosphoinositide hydrolysis or InsPs accumulation. Pertussis toxin pretreatment antagonized the response to PGE2, whereas 8-bromo cyclic AMP was without effect. The PGE2-induced decrease in InsPs may contribute to the suppression of insulin secretion. PMID- 2549956 TI - 2-Aminoethylarsonic acid as an analogue of ethanolamine phosphate. Endowment of ethanolamine-phosphate cytidylyltransferase with CTP pyrophosphatase activity. AB - 2-Aminoethylarsonic acid was tested for its ability to act as a substrate for ethanolamine-phosphate cytidylytransferase as a cytidylyl acceptor in place of ethanolamine phosphate. The expected product, like all mixed anhydrides of arsonic acids, should hydrolyse spontaneously with regeneration of the substrate analogue and CMP formation; such CMP production was observed. The limiting velocity with aminoethylarsonic acid is about 90% that with ethanolamine phosphate, and the Michaelis constant is below 20 mM. PMID- 2549957 TI - The mechanism of fungal cellulase action. Synergism between enzyme components of Penicillium pinophilum cellulase in solubilizing hydrogen bond-ordered cellulose. AB - Studies on reconstituted mixtures of extensively purified cellobiohydrolases I and II and the five major endoglucanases of the fungus Penicillium pinophilum have provided some new information on the mechanism by which crystalline cellulose in the form of the cotton fibre is rendered soluble. It was observed that there was little or no synergistic activity either between purified cellobiohydrolases I and II, or, contrary to previous findings, between the individual cellobiohydrolases and the endoglucanases. Cotton fibre was degraded to a significant degree only when three enzymes were present in the reconstituted enzyme mixture: these were cellobiohydrolases I and II and some specific endoglucanases. The optimum ratio of the cellobiohydrolases was 1:1. Only a trace of endoglucanase activity was required to make the mixture of cellobiohydrolases I and II effective. The addition of cellobiohydrolases I and II individually to endoglucanases from other cellulolytic fungi resulted in little synergistic activity; however, a mixture of endoglucanases and both cellobiohydrolases was effective. It is suggested that current concepts of the mechanism of cellulase action may be the result of incompletely resolved complexes between cellobiohydrolase and endoglucanase activities. It was found that such complexes in filtrates of P. pinophilium or Trichoderma reesei were easily resolved using affinity chromatography on a column of p-aminobenzyl-1-thio-beta-D-cellobioside. PMID- 2549958 TI - Identification of the ATP + Mg-dependent and polycation-stimulated protein phosphatases in the germinal vesicle of the Xenopus oocyte. AB - Two protein phosphatase activities were characterized in the germinal vesicle of Xenopus laevis oocytes after manual dissection of the nucleus. One enzyme can be classified as an active form of the ATP + Mg-dependent (AMD) phosphatase, the other as a polycation-stimulated (PCS) phosphatase. The activity of the PCS phosphatase is localized exclusively in the soluble compartment of the nucleus (nucleoplasm). The catalytic subunit of the AMD phosphatase activity is associated either with the nuclear particulate fraction or with an inhibitory subunit in the nucleoplasm. PMID- 2549960 TI - The 'methylamine oxidase' system of an obligate methylotroph. AB - The terminal respiratory oxidase was solubilized from membranes of organism 4025, an obligate methylotroph. The partially purified oxidase is probably a cytochrome co. It does not oxidize amicyanin, but it oxidizes 'azurin' and cytochromes cH and cL. By using a complete 'methylamine oxidase' system reconstituted from pure methylamine dehydrogenase, purified oxidase and soluble blue copper proteins and cytochromes, it was confirmed that amicyanin is essential for methylamine oxidation; it could not be replaced by 'azurin' or cytochrome cH or cL. It was shown that the usual mediator between amicyanin and the oxidase is cytochrome cH, with 'azurin' able to replace it during growth at the high copper concentrations required for optimum growth of this unusual methylotroph. PMID- 2549959 TI - Genetic regulation of beta 2-adrenergic receptors in 3T3-L1 fibroblasts. AB - The beta 2-adrenergic receptor from mouse 3T3-L1 cells is up-regulated through genetic mechanisms by glucocorticoids and butyrate. To study the genetic regulation of these receptors, we sequenced a 5 kb region of genomic DNA from 3T3 L1 cells, containing the beta-adrenergic receptor gene and approx. 1.5 kb of both 5' and 3' flanking sequences. The sequence contained one copy of an 8 bp consensus sequence which can confer phorbol ester-responsiveness to genes. Phorbol esters attenuated the up-regulation of beta 2-adrenergic receptors by glucocorticoids but not by butyrate. This effect was probably due to a phorbol ester-induced decrease in glucocorticoid receptor number. Using methylation sensitive restriction enzymes, we examined the methylation of a CG-rich region occurring 5' to the gene and did not detect any changes in methylation of this region upon dexamethasone or butyrate treatment. A total of 16 putative glucocorticoid response elements were found which may mediate the glucocorticoid induced increase in beta 2-adrenergic receptors. A comparison of the regulatory sequences of the two beta-adrenergic receptor subtypes from human and mouse confirms the observed physiological controls of receptor subtype expression and offers an explanation as to why the subtypes differ in genetic regulation. PMID- 2549961 TI - Facile preparation and characterization of the toxin from Bacillus thuringiensis var. kurstaki. AB - We report a simple three-step method of generating a homogeneous toxic fragment (toxin) in high yield from B. thuringiensis var. kurstaki. Purified crystals were digested with trypsin at pH 10.5, followed by (NH4)2SO4 precipitation and dialysis. For the HD73 strain the preparation is toxic to eastern-spruce-budworm (Choristoneura fuminiferana) larvae. It gives a single 66 kDa band on polyacrylamide-gel electrophoresis and a single band with an isoelectric point of 5.5 on an isoelectric-focusing gel. A single isoleucine N-terminus was detected, and the first 20 amino acids were found to be identical with those predicted from the gene nucleotide sequence. A single lysine C-terminus was detected, and the amino acid composition was in excellent agreement with tryptic cleavages at arginine-28 and lysine-623 of the protoxin. Raman spectroscopic analysis gave values of 20% alpha-helix, 35% beta-sheet and 45% unordered structure. The resistance of the toxin to most proteinases and its susceptibility to proteolysis by papain and Pronases indicates a compact multidomain structure. PMID- 2549962 TI - Inhibition of collagen hydroxylation by 2,7,8-trihydroxyanthraquinone in embryonic-chick tendon cells. AB - Prolyl 4-hydroxylase (EC 1.14.11.2) is an essential enzyme in the post translational modification of collagen. Inhibitors of this enzyme are of potential interest for the treatment of diseases involving excessive deposition of collagen. 2,7,8-Trihydroxyanthraquinone (THA) is an effective inhibitor of prolyl 4-hydroxylase by virtue of its ability to compete with the co-substrate 2 oxoglutarate (Ki = 40.3 microM). Using a simple and reproducible assay for collagen hydroxylation, we show that THA inhibits the hydroxylation of collagen in embryonic-chick tendon cells in short-term culture, with an IC50 value (concn. giving 50% inhibition) of 32 microM. In comparison, the ethyl ester of 3,4 dihydroxybenzoic acid has an IC50 value of 0.1 mM against collagen hydroxylation by chick tendon cells, whereas its Ki versus isolated prolyl 4-hydroxylase is 49 microM. PMID- 2549963 TI - Purification and characterization of two high-density-lipoprotein-binding proteins from rat and human liver. AB - The liver plays a major role in the metabolism of plasma high-density lipoprotein (HDL). Several groups have postulated, but others refuted, the existence of a classical membrane receptor which recognizes HDL. In the present study, we identified and purified two liver HDL-binding proteins of 120 kDa (HB1) and 100 kDa (HB2), with apparent specificity for HDL3 devoid of E apolipoprotein. The plasma membrane was the richest source of the HDL-binding protein. Both proteins bound A-I and A-II apolipoproteins and retained HDL-binding activity after final purification. HB1 activity, but not that of HB2, was lost after treatment with beta-mercaptoethanol, but reduction did not change the apparent molecular mass of either protein. Antibodies against HB1 or HB2 did not cross-react, and preliminary structural investigations provide evidence to suggest that HB1 and HB2 are not structurally related. We thus provide evidence for at least two liver plasma-membrane proteins which bind HDL apolipoproteins, suggesting that protein protein interaction participates to some degree in the mechanism of HDL recognition by cells. PMID- 2549964 TI - Carbachol and histamine stimulation of guanine-nucleotide-dependent phosphoinositide hydrolysis in rat brain cortical membranes. AB - Guanine nucleotides have been shown to stimulate phosphoinositide breakdown in brain membranes, but no potentiation of such an effect by agonist was demonstrated. We have studied the effect of carbachol and histamine on guanosine 5'-[gamma-thio]triphosphate (GTP[S]) stimulation of inositol phosphates formation in [3H]inositol-labelled rat brain cortical membranes. In this preparation, GTP[S] enhancement of phosphoinositide hydrolysis required the presence of MgATP and low Ca2+ concentration (100 nM). Carbachol potentiation of the GTP[S] effect was only observed when 1 mM-deoxycholate was also added. Under these conditions, stimulated production of [3H]inositol phosphates was linear for at least 15 min, and [3H]inositol bisphosphate [( 3H]IP2) accounted for approx. 80%, whereas the amount of [3H]inositol trisphosphate [( 3H]IP3) was very low. Stimulation by GTP[S] was concentration-dependent (half-maximal effect at 0.86 microM), and its maximal effect (815% over basal) was increased by 1 mM-carbachol (1.9-fold) and histamine (1.7-fold). Both agonists decreased the slope index of the GTP[S] concentration/effect curve to values lower than unity, suggesting the appearance of some heterogeneity in the population of guanine-nucleotide-binding proteins (G proteins) involved. The carbachol and histamine effects were also concentration dependent, and were inhibited by atropine and mepyramine respectively. Fluoroaluminate stimulated phosphoinositide hydrolysis to a higher extent than GTP[S] plus carbachol, and these stimulations were not additive, indicating that the same polyphosphoinositide phospholipase C-coupled G-protein mediates both effects. PMID- 2549965 TI - Synthesis of nitric oxide from L-arginine by neutrophils. Release and interaction with superoxide anion. AB - Stimulated rat peritoneal neutrophils release a platelet inhibitory factor with the pharmacological properties of NO. This release is inhibited by NG-monomethyl L-arginine and L-canavanine, indicating that it occurs through a mechanism similar to that in vascular endothelial cells and macrophages. As the degree of stimulation increases, the factor released is progressively inactivated by concomitant release of superoxide anions. PMID- 2549966 TI - Solubilization of rat liver vasopressin receptors as a complex with a guanine nucleotide-binding protein and phosphoinositide-specific phospholipase C. AB - Vasopressin V1 receptors were solubilized from rat liver plasma membranes with the detergent lysophosphatidylcholine. [[3H]Arginine]vasopressin (AVP) binding to the solubilized preparations was specific and saturable, with a dissociation constant of 0.6 nM. Cross-linking of [125I]vasopressin to the solubilized fraction, studied by SDS/polyacrylamide-gel-electrophoretic analysis, demonstrated the presence of a 65 kDa band which was specifically labelled with [125I]vasopressin. Specific binding of [3H]AVP to these solubilized receptors was decreased by guanine nucleotides, but not by adenosine 5'-[beta gamma imido]triphosphate. Addition of vasopressin increased specific binding of 35S labelled guanosine 5'-[gamma-thio]triphosphate (GTP[35S]) to the solubilized fractions, indicating co-solubilization of GTP-binding protein(s) [G-protein(s)] and vasopressin receptors. The solubilized fraction was insensitive to both cholera- and pertussistoxin treatment. Immunoblotting of the solubilized fraction with antibodies specific for a phosphoinositide-specific phospholipase C (PI-PLC I) demonstrated the presence of a 60 kDa protein. Anti-PI-PLC I antiserum immunoprecipitated solubilized vasopressin-binding sites from rat liver (V1), but not solubilized vasopressin-binding sites from hog kidney (V2). Similar results were obtained with an anti-PI-PLC I IgG affinity column. The solubilized (V1) receptors were enriched by ion-exchange and high-performance gel-filtration liquid chromatography. Vasopressin-binding activity was co-eluted with PI-PLC I and GTP[S]-binding activity on a DEAE-Sepharose column. The major vasopressin- and GTP[35S]-binding activities were co-eluted with PI-PLC I activity at approx. 240 kDa suggesting that vasopressin receptors from rat liver membranes can be solubilized as a complex of receptor-coupler-effector by using the detergent lysophosphatidycholine. PMID- 2549967 TI - Metabolism of inositol phosphates in alpha 1-adrenoceptor-stimulated and homogenized cardiac myocytes of adult rats. AB - Previous studies demonstrated the accumulation of inositol mono- and poly phosphates in carbamoylcholine-stimulated cultured cardiac ventricular myocytes of adult rats [Berg, Guse & Gercken (1989) Biochim. Biophys. Acta 1010, 100-107]. Stimulation with noradrenaline (50 microM) in the presence of propranolol (10 microM) led to a time-dependent pattern of inositol mono- and poly-phosphates in cultured cardiac-ventricular myocytes. Ins(1,4,5)P3 and Ins(1,3,4,5)P4 increased in a rapid initial phase. The degradation products of Ins(1,4,5)P3, namely Ins(1,4)P2 and Ins(4)P, accumulated between 1 and 15 min. Ins(1,3,4,5)P4 was rapidly dephosphorylated to Ins(1,3,4)P3, which was then metabolized to Ins(1,3)P2 and Ins(3,4)P2. The last two InsP2 isomers and their degradation products, Ins(1)P and Ins(3)P (determined as an enantiomeric mixture), increased at extended stimulation periods. To confirm the pathway of Ins(1,3,4,5)P4 degradation, homogenates of isolated ventricular myocytes were incubated with [3H]INs(1,3,4,5)P4. The subsequent products were Ins(1,3,4)P3, Ins(3,4)P2, Ins(1,3)P2 and InsP. The effect of noradrenaline was antagonized by prazosin (0.1 microM), but not by yohimbine (0.1 microM), indicating phosphoinositidase activation via the alpha 1-adrenoceptor. PMID- 2549969 TI - Interaction of cerebral-cortical membranes with exogenously added phosphatidylinositol 4,5-bisphosphate. Effects on measured phospholipase C activity. AB - Exogenously added phosphatidylinositol 4,5-bisphosphate (PtdInsP2) is rapidly associated with cerebral-cortical membranes. Substrate association with membranes was promoted by Mg2+, but inhibited by bivalent chelators. Once associated with the membrane, the PtdInsP2 was resistant to displacement by EDTA. The apparent phospholipase C activity was dependent on the degree of association of substrate with membranes. After preincubation of membranes with substrate, PtdInsP2 hydrolysis was independent of the incubation volume, indicating that substrate and membrane-associated phospholipase C were not independently diluted. Hydrolysis of the membrane-associated substrate was stimulated by Ca2+, guanosine 5'-[beta gamma-imido]triphosphate (p[NH]ppG), guanosine 5'[gamma thio]triphosphate and carbachol in the presence of p[NH]ppG. Carbachol in the absence of guanine nucleotides, GDP, GTP, ATP and pyrophosphate was ineffective. These results demonstrate that exogenously added PtdInsP2 substrate is rapidly associated with membranes and hydrolysed by a phospholipase C whose activity is regulated by guanine nucleotides and agonist in the presence of guanine nucleotides. Use of exogenously added substrate for studies on the regulation of membrane phospholipase C requires consideration as to possible effects of incubation conditions on the partitioning of substrate into membranes. PMID- 2549968 TI - Proteolytic processing of a coleopteran-specific delta-endotoxin produced by Bacillus thuringiensis var. tenebrionis. AB - Insecticidal protein delta-endotoxin crystals harvested from sporulated cultures of Bacillus thuringiensis var. tenebrionis contain a major polypeptide of 67 kDa and minor polypeptides of 73, 72, 55 and 46 kDa. During sporulation, only the 73 kDa polypeptide could be detected at stage I. The 67 kDa polypeptide was first detected at stage II and increased in concentration throughout the later stages of sporulation and after crystal release, with a concomitant decrease in the 73 kDa polypeptide. This change could be blocked by the addition of proteinase inhibitors. Trypsin or insect-gut-extract treatment of the delta-endotoxin crystals after solubilization resulted in a cleavage product of 55 kDa with asparagine-159 of the deduced amino acid sequence of the toxin [Hofte, Seurinck, van Houtven & Vaeck (1987) Nucleic Acids Res. 15, 71-83; Sekar, Thompson, Maroney, Bookland & Adang (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 7036-7040; McPherson, Perlak, Fuchs, Marrone, Lavrik & Fischhoff (1988) Biotechnology 6, 61 66] at the N-terminus. This polypeptide was found to be as toxic in vivo as native delta-endotoxin. PMID- 2549970 TI - Reversible thermal inactivation of the quinoprotein glucose dehydrogenase from Acinetobacter calcoaceticus. Ca2+ ions are necessary for re-activation. AB - The soluble form of the homogeneous quinoprotein glucose dehydrogenase from Acinetobacter calcoaceticus is reversibly inactivated at temperatures above 35 degrees C. An equilibrium is established between active and denatured enzyme, this depending on the protein concentration and the inactivation temperature used. Upon thermal inactivation the enzyme dissociates into the prosthetic group pyrroloquinoline quinone and the apo form of glucose dehydrogenase. After inactivation at 50 degrees C active enzyme is re-formed again at 25 degrees C. Ca2+ ions are necessary for the re-activation process. The velocity of re activation depends on the protein concentration, the concentration of the prosthetic group pyrroloquinoline quinone and the Ca2+ concentration. The apo form of glucose dehydrogenase can be isolated, and in the presence of pyrroloquinoline quinone and Ca2+ active holoenzyme is formed. Even though native glucose dehydrogenase is not inactivated in the presence of EDTA or trans-1,2 diaminocyclohexane-NNN'NH-tetra-acetic acid, Ca2+ stabilizes the enzyme against thermal inactivation. Two Ca2+ ions are found per subunit of glucose dehydrogenase. The data suggest that pyrroloquinoline quinone is bound at the active site via a Ca2+ bridge. Mn2+ and Cd2+ can replace Ca2+ in the re activation mixture. PMID- 2549971 TI - The cytochromes of anaerobically grown Escherichia coli. An electron-paramagnetic resonance study of the cytochrome bd complex in situ. AB - The e.p.r. signals attributable to a cytochrome bd-type ubiquinol:O2 oxidoreductase (cytochrome b-558-b-595-d) were studied in a cytoplasmic membrane preparation of Escherichia coli that had been grown on glycerol with fumarate as respiratory-chain oxidant. Two major high-spin ferric haem signals were resolved on the basis of their potentiometric behaviour: a rhombic high-spin species (gx = 6.25, gy = 5.54) was assigned to haem b-595, and an axial high-spin (gx = 5.97, gy = 5.96) species was assigned to the haem d. These signals titrated with Em.7 values of 154 and 261 mV respectively, corresponding closely to optically determined values for haem b-595 and haem d. At high potentials (greater than 300 mV) the rhombic species attributable to haem b-595 underwent a partial transition to a second rhombic species with g-values of 6.24 (gx) and 5.67 (gy). The high spin ferric haem spectra were affected by O2, CO, cyanide and pH. A low-spin ferric haem signal was observed at g = 3.3 (gz), which titrated with an Em.7 of 226 mV, and this was assigned to haem b-558. The data support a model for cytochrome bd with two ligand-binding sites, a single haem d and a single haem b 595. PMID- 2549972 TI - Polymyxin B diminishes blood flow to brown adipose tissue and lactating mammary gland in the rat. Possible mechanism of its action to decrease the stimulation of lipogenesis on refeeding. AB - Polymyxin B, a cyclic decapeptide antibiotic, increased blood glucose and lactate, and inhibited the stimulation of lipogenesis in interscapular brown adipose tissue and lactating mammary gland of starved-refed virgin and lactating rats respectively. Lipogenesis was not inhibited in white adipose tissue or liver. The antibiotic increased the haematocrit. The relative blood flow to brown adipose tissue and lactating mammary gland was decreased by polymyxin B, and this was accompanied by a decrease in tissue ATP content. In vitro polymyxin B did not affect glucose utilization or conversion into lipid, nor the stimulation by insulin of these processes in brown-adipose-tissue slices. Treatment of rats in vivo with polymyxin B resulted in decreased utilization of glucose in vitro in brown-adipose-tissue slices. Similarly, acini from mammary glands of polymyxin B treated lactating rats had decreased rates of conversion of [1-14C]glucose to lipid. It is concluded that the effects of polymyxin B may be brought about by decreases in tissue blood flow. The possibility that these effects are secondary to inhibition of glucose utilization cannot be ruled out. PMID- 2549973 TI - Purification of bovine brain inositol 1,4,5-trisphosphate 3-kinase. Identification of the enzyme by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. AB - Inositol 1,4,5-trisphosphate (InsP3) 3-kinase catalyses the ATP-dependent phosphorylation of InsP3 to inositol 1,3,4,5-tetrakisphosphate (InsP4). A method is presented for the rapid purification of InsP3 3-kinase from bovine brain by calmodulin (CaM)-Sepharose affinity chromatography. Maximal activation of the purified InsP3 3-kinase by Ca2+/CaM was 6-7-fold as compared with the activity measured in the presence of EGTA (1 mM) and 10 microM-InsP3. At 10 microM-InsP3 and 0.1 mM free Ca2+, half-maximal activation required about 2 nM-CaM. The mechanism of activation by CaM appeared to be an increase in the maximal velocity of the enzyme without a substantial change in the Km for InsP3. Further purification was achieved by phosphocellulose chromatography eluted with ATP. Specific activity of the purified enzyme at 37 degrees C and 10 microM-InsP3 was 10-20 mumol/min per mg. The apparent Mr of the enzyme, determined by f.p.l.c.-gel filtration, was estimated as about 44,000. The purified InsP3 3-kinase was subjected to SDS/10%-polyacrylamide-gel electrophoresis. InsP3 3-kinase activity was associated with three silver-stained bands, which migrated with apparent Mr values of approx. 52,000, 38,000 and 35,000. PMID- 2549974 TI - Anomeric specificity of D-glucose phosphorylation by rat liver glucose-6 phosphatase. AB - The anomeric specificity of D-glucose phosphorylation by hepatic glucose-6 phosphatase was examined in rat liver microsomes incubated in the presence of carbamoyl phosphate. At 10 degrees C, the Km for the equilibrated hexose and phosphate donor was close to 56 mM and 11 mM, respectively. The enzymic activity, which was increased in diabetic rats, was about 40% lower in untreated than in sonicated microsomes. No anomeric difference in affinity was found in sonicated microsomes. In untreated microsomes, however, the Km for beta-D-glucose was slightly lower than that for alpha-D-glucose. The maximal velocity was higher with beta- than alpha-D-glucose in both untreated and sonicated microsomes. These data indicate that the phosphotransferase activity of glucose-6-phosphatase cannot account for the higher rate of glycolysis and glycogen synthesis found in hepatocytes exposed to alpha- rather than beta-D-glucose. PMID- 2549975 TI - 5'-Nucleotidase activity and adenosine formation in stimulated, hypoxic and underperfused rat heart. AB - Changes in 5'-nucleotidase activity were calculated on the basis of alterations in ATP, ADP, phosphocreatine, Pi, Mg2+, IMP and AMP, determined by using 31P n.m.r. spectroscopy and h.p.l.c., during isoprenaline infusion, graded hypoxia and graded underperfusion in isolated rat heart. Calculated activity changes were compared with the total efflux of purines (adenosine + inosine + hypoxanthine) in order to assess the involvement of various 5'-nucleotidases in formation of adenosine. Purine efflux exhibited an exponential relation with cytosolic [AMP] during isoprenaline infusion and hypoxia (r = 0.92 and 0.95 respectively), supporting allosteric activation of 5'-nucleotidase under these conditions. Purine efflux displayed a linear relation with cytosolic [AMP] during graded ischaemia (r = 0.96), supporting substrate regulation in the ischaemic heart. The calculated activities of membrane-bound ecto-5'-nucleotidase were similar to the observed relations between purine efflux and cytosolic [AMP] in all hearts. The calculated activities of the ATP-activated cytosolic and lysosomal enzymes and of the ATP-inhibited cytosolic 5'-nucleotidase could not explain the observed release of purines under the conditions examined. These results indicate that the kinetic characteristics of the membrane-bound ecto-enzyme are consistent with an important role in the formation of extracellular adenosine, whereas the characteristics of the other 5'-nucleotidases are inconsistent with roles in adenosine formation under the conditions of the present study. PMID- 2549976 TI - Low-density-lipoprotein receptors in different rabbit liver cells. AB - Receptor-dependent uptake mechanisms for low-density lipoprotein (LDL) were studied in rabbit liver parenchymal and non-parenchymal cells. Hybridization studies with a cDNA probe revealed that mRNA for the apo (apolipoprotein) B,E receptor was present in endothelial and Kupffer cells as well as in parenchymal cells. By ligand-blotting experiments we showed that apo B,E-receptor protein was present in both parenchymal and non-parenchymal cells. Studies of binding of homologous LDL in cultured rabbit parenchymal cells suggested that about 63% of the specific LDL binding was mediated via the apo B,E receptor. Approx. 47% of the specific LDL binding was dependent on Ca2+, suggesting that specific Ca2+ dependent as well as Ca2+-independent LDL-binding sites exist in liver parenchymal cells. Methylated LDL bound to the parenchymal cells in a saturable manner. Taken together, our results showed that apo B,E receptors are present in rabbit liver endothelial and Kupffer cells as well as in the parenchymal cells, and that an additional saturable binding activity for LDL may exist on rabbit liver parenchymal cells. This binding activity was not inhibited by EGTA or reductive methylation of lysine residues in apo B. LDL degradation in parenchymal cells was mainly mediated via the apo B,E receptor. PMID- 2549977 TI - Uptake, production and metabolism of cysteinyl leukotrienes in the isolated perfused rat liver. Inhibition of leukotriene uptake by cyclosporine. AB - 1. The isolated perfused rat liver efficiently takes up cysteinyl leukotrienes (LTs) C4, D4, E4 and N-acetyl-LTE4 from circulation. More than 70% of these cysteinyl LTs are excreted from liver into bile within 1 h of onset of a 5 min infusion, while about 5% remain in the liver. About 20% of infused N-acetyl-LTE4 escapes hepatic first-pass extraction under our conditions. 2. Metabolites of LTC4 appearing in bile within 20 min of the onset of infusion include mainly LTD4 and N-acetyl-LTE4, but also omega-hydroxy-N-acetyl-LTE4 and omega-carboxy-N acetyl-LTE4. Metabolites generated from omega-carboxy-N-acetyl-LTE4 by beta oxidation from the omega-end represent the major biliary LTs secreted at later times. 3. Stimulation of the isolated perfused liver by the combined infusion of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) and the Ca2+ ionophore A23187 results in a transient increase of endogenous cysteinyl LT production, which is independent of extrahepatic cells. 4. The immunosuppressive drug cyclosporine causes a dose-dependent inhibition of hepatobiliary cysteinyl LT excretion, probably by interference with the sinusoidal uptake system for cysteinyl LTs. PMID- 2549978 TI - Changes in the activities of adenosine-metabolizing enzymes in six regions of the rat brain on chemical induction of hypothyroidism. AB - 1. Rats (4 weeks old) were made hypothyroid by treatment with propylthiouracil and a low-iodine diet for a further period of 4 weeks. Synaptosomal membranes, myelin and 105,000 g soluble fractions were obtained from six regions of the brain. 2. Hypothyroidism resulted in 2-5-fold increases in membrane-bound 5' nucleotidase activity in synaptosomal fractions obtained from cerebellum, cortex, striatum and hippocampus. By contrast, myelin 5'-nucleotidase activity was slightly increased only in the medulla oblongata. 3. Hypothyroidism did not change adenosine deaminase activity, but decreased adenosine kinase activity by approx. 40% in soluble fractions obtained from cerebellum, hippocampus, striatum and hypothalamus. 4. It is suggested that these changes in hypothyroidism, in particular the increases in 5'-nucleotidase activity, could enhance the neuromodulatory effect of adenosine to decrease neurotransmitter release. PMID- 2549979 TI - The effects of Triton X-100 and chlorpromazine on the Mg2+-dependent and Mg2+ independent phosphatidate phosphohydrolase activities of rat lung. AB - Lung contains both Mg2+-dependent and Mg2+-independent phosphatidate phosphohydrolase activities. Addition of Triton X-100 (0.5%) or chlorpromazine (1 mM) leads to a marked increase in the total phosphatidate phosphohydrolase activity in rat lung microsomes (microsomal fractions), but a decrease in the Mg2+-dependent activity. These observations suggest that the Mg2+-independent activity is stimulated, whereas the Mg2+-dependent activity is inhibited. However, the possibility exists that Triton X-100 could stimulate the Mg2+ dependent enzymic activity in an Mg2+-independent manner. In addition, the positively charged amphiphilic drug could be replacing the enzyme's requirement for Mg2+. These two possibilities were examined by using subcellular fractions in which the Mg2+-dependent phosphatidate phosphohydrolase had been abolished by heat treatment at 55 degrees C for 15 min. Heat treatment does not affect the microsomal Mg2+-independent phosphohydrolase to any great extent. Since the 6-8 fold stimulations due to Triton X-100 and chlorpromazine are retained after heat treatment of this fraction, the Mg2+-independent activity must be involved. Addition of Triton X-100 and chlorpromazine to cytosol virtually abolishes the Mg2+-dependent phosphatidate phosphohydrolase activity and decreases the Mg2+ independent activity by half. Heat treatment also abolishes the Mg2+-dependent activity and decreases the Mg2+-independent activity by over half. The Mg2+ independent phosphatidate phosphohydrolase activity remaining after heat treatment was not affected by Triton X-100 or chlorpromazine. These studies demonstrate that Triton X-100 and chlorpromazine specifically stimulate the heat stable Mg2+-independent phosphatidate phosphohydrolase activity in rat lung microsomes. In contrast, the heat-labile Mg2+-independent phosphatidate phosphohydrolase activities in cytosol are inhibited by these reagents. Triton X 100 and chlorpromazine inhibit the Mg2+-dependent phosphatidate phosphohydrolase activities in both rat lung microsomes and cytosol. These results are consistent with the view that a single Mg2+-dependent phosphatidate phosphohydrolase present in both microsomes and cytosol is specifically involved in glycerolipid metabolism. PMID- 2549980 TI - Herpes simplex virus-1-specific proteins are involved in alteration of polyphosphoinositide metabolism in baby-hamster kidney cells. AB - Herpes simplex virus type 1 (HSV-1) induces altered phosphoinositide metabolism in baby hamster kidney (BHK) cells, measured as incorporation of [3H]inositol or [32P]Pi [Langeland, Haarr & Holmsen (1986) Biochem. J. 237, 707-712]. We now report that this response in the inositol phospholipids is dependent on virus specific proteins synthesized in the beta (early) stage of virus protein synthesis. This was demonstrated both by resistance to the inhibitory effect of cycloheximide after this stage of infection, and by the use of temperature sensitive (ts) mutants of HSV-1; ts mutants in which protein synthesis was blocked so that only the alpha proteins were expressed showed a PIP2/PIP (phosphatidylinositol 4,5-bisphosphate/phosphatidylinositol 4-monophosphate) ratio similar to uninfected cells, while ts mutants which were defective in protein synthesis at a late beta stage or later showed increased PIP2/PIP ratios similar to cells infected by wild type HSV-1. PMID- 2549982 TI - DNA topoisomerase I from Diplococcus pneumoniae. AB - A type I topoisomerase has been purified from Diplococcus pneumoniae using phosphocellulose and hydroxylapatite chromatography. The purified enzyme catalyses the relaxation of negatively supercoiled DNA. The relaxation requires Mg2+ and is favoured by 0.2 M monovalent cations. The enzyme does not exhibit catenating or supercoiling activities. Using circular pBR322 DNA from dam+- and dam- -hosts as substrates for the enzyme, the relaxation reaction proceeds with somewhat higher efficiency with plasmids containing methylated adenine in GATC sequences. Plasmids from dcm+- and dcm- -hosts show no difference in reactivity. PMID- 2549981 TI - Approaches to characterize by density-fractionation age-dependent properties of reticulocyte mitochondria. AB - Mitochondria from rabbit reticulocytes of varying maturity were prepared by means of two different methods and separated in sucrose and Percoll density gradients. A mechanical method of cell disintegration under isotonic conditions followed by centrifugation in a Percoll density gradient proved to be adequate to reflect the physiological conditions. "Young" mitochondria exhibit in this system greater density and higher specific activity of cytochrome oxidase. PMID- 2549983 TI - SRP-dependent membrane integration of the beta-subunit of Na+,K+-ATPase. AB - cDNA clones coding for either full-length or truncated forms of the beta-subunit of the Na+,K+-ATPase from pig kidney were engineered into a transcription vector based on a T7 promotor. In vitro transcription and subsequent translation of the mRNA in the presence of rough microsomes (RM) yielded beta-subunit molecules that were N-glycosylated and correctly inserted into the membrane. The signal peptide was not cleaved off. This membrane integration was found to be dependent on the function of the signal recognition particle (SRP). Several lines of evidence suggest that the hydrophilic aminoterminal domain of 34 amino acid residues preceding the postulated signal sequence is located on the cytoplasmic side whereas the carboxyterminal glycosylated domain is located on the exoplasmic side of the ER (endoplasmic reticulum)-membrane (type II membrane protein). PMID- 2549984 TI - Expression of active alpha-3 subunit of rat brain Na+,K+-ATPase from the messenger RNA injected into Xenopus oocytes. AB - Cloned cDNA encoding the so-far uncharacterized alpha-3 subunit of rat brain Na+,K+-ATPase (Hara et al. (1987) J. Biochem. 102, 43-58, Shull et al. (1986) Biochemistry 25, 8125-8132) was incorporated into a vector carrying the SP6 promoter. The mRNA produced in vitro was injected into Xenopus oocytes with the mRNA encoding the Na+,K+-ATPase beta subunit of Torpedo electroplax. Increased Na+,K+-ATPase activity in the oocyte membrane was observed. This newly expressed activity was inhibited by ouabain (Ki = 1.5 x 10(-7) M), suggesting that the alpha-3 subunit of rat brain Na+,K+-ATPase is a highly ouabain-sensitive catalytic subunit. PMID- 2549985 TI - Expression of rat protein phosphatase 2C (IA) in Escherichia coli. AB - A cDNA containing the entire coding sequence of rat type 2C (IA) protein phosphatase was expressed in Escherichia coli. An extract of bacterial cells harboring the recombinant plasmid contained a major (Mr = 41,000 - 43,000) and a minor (Mr = 30,000) protein band; both of these reacted with an anti-type 2C protein phosphatase serum. The size of the major protein band agrees well with that of the 2C phosphatase conceptualized from the cognate cDNA. A Mg2+-dependent protein phosphatase activity was detected in extracts containing the recombinant protein, but not in host cell extracts. Based on these results, it is concluded that the isolated cDNA clone encodes a functional type 2C protein phosphatase. PMID- 2549986 TI - Competitive inhibition of tritium-labeled platelet-activating factor binding to rabbit platelet membranes by amiloride and amiloride analogs. AB - Amiloride and its structural analogs, ethylisopropyl amiloride, benzamil, and dichlorobenzamil, inhibit both the specific [3H]C18-PAF binding to rabbit platelet membranes and PAF-induced aggregation of gel-filtered rabbit platelets. Detailed analysis of binding inhibitions demonstrate that ethylisopropyl amiloride is a competitive inhibitor with an equilibrium dissociation constant (KB) of 23 microM. The concentration of amiloride and its analogs needed to inhibit the PAF-induced aggregation is high and there exists no correlation between their inhibitory activities of platelet aggregation and those of Na+/H+ antiporter. However, the inhibitory effects on the PAF-induced aggregation are parallel to those on the specific [3H]C18-PAF binding. The inhibitory effects of amiloride and its analogs on the activation of platelets are at the PAF-receptor binding step, rather than at the Na+/H+ antiporter. PMID- 2549987 TI - A rapid solution immunoassay to quantify binding of the human immunodeficiency virus envelope glycoprotein to soluble CD4. AB - We developed a particle concentration fluorescent immunoassay to quantify the binding in solution of the human immunodeficiency virus (HIV) external glycoprotein (gp120) to soluble CD4 (sCD4). The assay is rapid (1 hr), quantitative, and requires as little as 0.1 pmole of gp120 per evaluation. We find that gp120, purified from recombinant baculovirus infected insect cells, is suitable for the assay. Moreover, sCD4s obtained either from recombinant E. coli or mammalian cells, consisting of the N-terminal two domains (about 180 amino acids) as well as linked to the active regions of Pseudomonas exotoxin A, bind gp120 similarly. PMID- 2549988 TI - Cyclic and noncyclic inositol phosphates are formed at different ratios by phospholipase C isozymes. AB - The cyclic inositol phosphate content in the product of PLC-beta, gamma, and delta mediated cleavage of three phosphoinositides, PtdIns, PtdIns(4)p, and PtdIns(4,5)P2, was measured under several different experimental conditions. The ratio of cyclic to noncyclic product generally decreased in the order PLC-beta greater than PLC-delta greater than PLC-gamma. For all three enzymes the ratio decreased in the order PtdIns greater than PtdIns(4)P greater than PtdIns(4,5)P2. For all combinations of the three enzymes and three substrates cyclic product content was always higher at pH 5.5 than at pH 7.0. The effect of Ca2+ on the ratio of cyclic to noncyclic was also measured. The ratio remained constant between 0.5 microM and 2 mM for PtdIns. For PtdIns(4)P and PtdIns(4,5)P2, the ratios were unchanged between 0.5 and 500 microM, but increased abruptly at millimolar Ca2+ concentrations. PMID- 2549989 TI - Sodium-hydrogen exchange system in brush border membranes from cortical and medullary regions of the proximal tubule. AB - The Na+/H+ exchange system was studied in brush border membrane vesicles isolated from cortical and medullary regions of the proximal tubule of rabbit kidney. The activity of the exchanger was assessed by measuring hydrogen influx (monitored by acridine orange fluorescence), 22 Na influx and the sensitivity of these fluxes to amiloride and its analogue ethylisopropyl amiloride. In contrast to previously published data (indicating the absence of pH-gradient driven and amiloride sensitive 22Na-influx in medullary site vesicles (13, 15], Na+/H+ exchange activity could be detected in both membrane preparations by sodium tracer and fluorescence detection of hydrogen influx. Amiloride inhibition of 22Na influx was more effectively protected by increasing sodium concentration in cortical than in medullary vesicles, suggesting differences in the action of amiloride in these preparations. PMID- 2549990 TI - Electron transfer between liposomal cytochrome c1 and cytochrome c: catalytic implications of electrostatic potentials. AB - Kinetics measurements of the electron transfer between ferricytochrome c and liposomal ferrocytochrome c1 (with and without the hinge protein) were performed. The observed rate constants(kobs) of electron transfer between liposomal ferrocytochrome c1 and ferricytochrome c at different ionic strengths were measured in cacodylate buffer, pH 7.4, at 2 C. The effect of ionic strength on the rate constant(kobs) of electron transfer between liposomal cytochrome c1 and cytochrome c is far greater than that in the solution kinetics (Kim, C.H., Balny, C. and King, T.E. (1987) J. Biol. Chem. 262, 8103-8108). The result demonstrates that the membrane bound cytochrome c1 creates a polyelectrolytic microenvironment which appears to be involved in the control of electron transfer and can be modulated by the ionic strength. The involvement of electrostatic potentials in the electron transfer between the membrane bound cytochrome c1 and cytochrome c is discussed in accord with the experimental results and a polyelectrolyte theory. PMID- 2549992 TI - Oxygen dependent regulation of mammalian ribonucleotide reductase in vivo and possible significance for replicon initiation. AB - The intracellular concentration of the M2 specific free tyrosyl radical of ribonucleotide reductase in cultured Ehrlich ascites cells was estimated by EPR spectroscopy during imposition and after reversal of hypoxia. Under the same conditions, the intracellular intensity of CDP reduction was estimated indirectly by measuring the incorporation of radioactivity into DNA from labeled ribo- and deoxyribo-cytidine, respectively. The radical concentration distinctly decreased under hypoxia and reincreased upon reaeration. At the same time, the CDP reduction was greatly diminished and reactivated, respectively. These observations are interpreted in the sense of an O2 dependent regulation of the intracellular activity of ribonucleotide reductase. The O2 dependent deactivation and reactivation of the enzyme was temporally related to a specific suppression or a burst-like release of replicon initiations, respectively. Addition of 100 microM dCtd to hypoxic cells could substitute for reoxygenation with respect to triggering replicon initiations. A possible implication of the intracellular ribonucleotide reductase activity and the size of the dCTP pool in replicon initiation is discussed. PMID- 2549991 TI - Regulation of IGF-I receptors by corticotropin and angiotensin-II in cultured bovine adrenocortical cells. AB - The effects of angiotensin II (A-II) and corticotropin (ACTH) on insulin-like growth factor-I (IGF-I) receptors of bovine adrenocortical cells were investigated. Pretreatment of cells for 48 h with ACTH or A-II induced in a dose dependent manner an increase in [125I]IGF-I binding (ED50 congruent to 10(-11)M, Vmax = 10(-10) M with ACTH; ED50 congruent to 3.10(-9) M, Vmax = 10(-7) M with A II). This resulted from an increase in the number of binding sites without modification of the binding affinity. Pretreatment with 8-Bromo-cAMP (10(-3) M), a phorbol ester (PMA 10(-7) M) + ionophore A23187 (10(-7) M) produced a positive regulation of IGF-I receptors. Glucocorticoids did not mediate the effect of A-II and ACTH on IGF-I receptors. Since previous studies have shown that IGF-I increased ACTH and A-II receptors the present data indicate the existence of a reciprocal positive trophic effect between IGF-I and the two hormones on the regulation of their specific membrane-bound receptors. PMID- 2549993 TI - Inhibition of prostaglandin E2 synthesis by a blocker of epithelial chloride channels. AB - Arginine-vasopressin (AVP) elicits a variety of responses in cultured rat mesangial cells, among them stimulation of prostaglandin biosynthesis and activation of Cl- channels. AVP produced an 11-fold increase over basal levels in prostaglandin E2 release from cultured mesangial cells. This response was completely inhibited by 25 microM indomethacin and 82 +/- 5% inhibited by 25 microM 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB) which is a potent blocker of epithelial Cl- channels. The IC50 for NPPB inhibition of prostaglandin E2 release was 8 microM. Indomethacin and NPPB at 25 microM also inhibited AVP stimulated cellular accumulation of prostaglandin E2 by 98% and 79 +/- 7% respectively. The inhibitory effect of NPPB was not due to interference with the cellular response to AVP since at 50 microM it did not block AVP-stimulated release of arachidonate metabolites from cells metabolically labeled with [3H] arachidonic acid. It is suggested that NPPB inhibition of prostaglandin E2 synthesis is at the cyclooxygenase level on the basis of its structural similarity to the fenamic acid type of cyclooxygenase inhibitors. PMID- 2549994 TI - Structural differences between the glucocorticoid, dioxin and oxysterol receptors from rat liver cytosol. AB - The rat hepatic glucocorticoid, dioxin and oxysterol receptors were subjected to high performance liquid chromatography on size-exclusion and anion-exchange columns. Both the glucocorticoid receptor and the dioxin receptor had a Stokes radius Rs approximately 7.5 nm, expected value for heteromeric complexes containing a dimer of the Mr approximately 90,000 heat shock protein, hsp90 (Rs approximately 7.0 nm). The oxysterol receptor represented a much smaller entity (Rs approximately 6.0 nm). When analyzed on a Mono Q anion-exchange column, the molybdate-stabilized glucocorticoid receptor and dioxin receptor eluted as single peaks at approximately 0.30 M and 0.26-0.28 M NaCl, respectively, whereas the oxysterol receptor represented a less negatively charged species (0.11-0.14 M NaCl). Following washing of the Mono Q column with molybdate-free buffer, the activated monomeric glucocorticoid receptor was detected (0.10-0.12 M NaCl). In contrast, no modification in the elution pattern of the dioxin receptor and the oxysterol receptor was observed. These data demonstrate differences in the physico-chemical properties of the glucocorticoid, dioxin and oxysterol receptors, respectively, which might reflect structural differences. PMID- 2549995 TI - Effects of endothelin on sodium transport mechanisms: potential role in cellular Ca2+ mobilization. AB - The effects of endothelin on cellular Ca2+ mobilization were examined in cultured rat vascular smooth muscle cells (VSMC). Endothelin (10(-8)M) induced a rapid transient increase of [Ca2+]i from 77 +/- 3 to 104 +/- 5 nM (p less than .05) in VSMC. Preincubation (60 min) with endothelin (2 x 10(-6)M) increased basal [Ca2+]i from 77 +/- 3 to 105 +/- 8 nM (p less than .05). Preincubation with endothelin also enhanced vasopressin (10(-7)M)-stimulated peak levels of [Ca2+]i (528 +/- 20 nM vs 969 +/- 21 nM, p less than .01). Endothelin (10(-7)M) induced an intracellular alkalinization (7.18 +/- 0.03 vs 7.37 +/- 0.04, p less than .01) which was blocked by pretreatment with amiloride. The biphasic effects of endothelin on [Ca2+]i were similar to those of an endogenous inhibitor of Na-K ATPase that we examined in a previous study. Therefore, we examined the effects of endothelin on Na-K-ATPase in an enzyme preparation from hog cerebral cortex. At high concentrations, endothelin (10(-5)M) inhibited Na-K-ATPase in vitro. Thus, endothelin may exert its vasoconstrictor effects at least in part via alterations of cellular Ca2+ mobilization in VSMC. While the rapid transient increase of [Ca2+]i appears to reflect intracellular Ca2+ mobilization, the sustained effect on [Ca2+]i may be related to an increase of intracellular sodium mediated by inhibition of Na-K-ATPase and/or more likely by stimulation of the Na+/H+-antiport. PMID- 2549996 TI - Adrenergic regulation of the uncoupling protein expression in foetal rat brown adipocytes in primary culture. AB - The adrenergic and T3 modulation of UCP expression in non-proliferative foetal brown adipocyte primary cultures were studied. The UCP in the cultured cells was determined by immunological detection of the protein and by quantification of the mitochondrial GDP-binding. Our results showed a relative increase of 65-75% in UCP levels and 60-80% in the mitochondrial GDP-binding capacity under beta adrenergic stimulatory conditions, while neither alpha 1-adrenergic agonists nor T3 showed an effect. PMID- 2549997 TI - High pressure liquid chromatography of cyclic nucleotide phosphodiesterase from purified human T-lymphocytes. AB - The cyclic nucleotide phosphodiesterase (EC 3.1.4.17) in extracts of purified human peripheral blood T-lymphocytes was examined by ion exchange high pressure liquid chromatography. Four peaks of activity were isolated. The first peak of activity selectively hydrolyzed cyclic GMP. The following 3 peaks of activity (Ia, IIa and IIIa) were selective for cyclic AMP. The selective low Km cyclic AMP phosphodiesterase inhibitor, Ro 20-1724 (d,1-1,4-[3-butoxy-4-methoxybenzyl]-2 imidazolidinone), did not inhibit the activity in Ia whereas it did inhibit the activity in IIa and IIIa (IC50 = 17 microM). The authors conclude that ion exchange high pressure liquid chromatography described in this communication is a useful method for the isolation of different forms of cyclic nucleotide phosphodiesterase activity from human T-lymphocytes. PMID- 2549998 TI - A cytosolic protein activator of cardiac sarcolemmal phosphoinositide phospholipase C. AB - Ca2+ dependent polyphosphoinositide phospholipase C (PLC) activity in cardiac sarcolemma hydrolyzed both endogenous and exogenous phosphatidylinositol 4 phosphate (PIP) and phosphatidylinositol 4,5-bisphosphate (PIP2) with an associated increase in inositol bisphosphate (IP2). Dialyzed cytosol and certain fractions of cytosol isolated by anion exchange or gel filtration chromatography activated sarcolemmal PLC activity by approx. 100%. The PLC activator eluted with an apparent molecular weight of 160 Kdal on a Sephacryl 300 column and was destroyed by heat or trypsin treatment. Exogenous 3H-PIP2 was not hydrolyzed by cytosolic fractions containing sarcolemmal PLC activator. These studies demonstrate that the polyphosphoinositide PLC in cardiac sarcolemma is regulated by a cytosolic protein. PMID- 2549999 TI - Identification and partial characterization of discrete apolipoprotein A containing lipoprotein particles secreted by human hepatoma cell line HepG2. AB - The purpose of this study was to identify the apolipoprotein A-containing lipoprotein particles produced by HepG2 cells. The apolipoprotein A-containing lipoproteins separated from apolipoprotein B-containing lipoproteins by affinity chromatography of culture medium on concanavalin A were fractionated on an immunosorber with monoclonal antibodies to apolipoprotein A-II. The retained fraction contained apolipoproteins A-I, A-II and E, while the unretained fraction contained apolipoproteins A-I and E. Both fractions were characterized by free cholesterol as the major and triglycerides and cholesterol esters as the minor neutral lipids. Further chromatography of both fractions on an immunosorber with monoclonal antibodies to apolipoprotein A-I showed that 1) apolipoprotein A-II only occurs in association with apolipoprotein A-I, 2) apolipoprotein A-IV is only present as part of a separate lipoprotein family (lipoprotein A-IV), and 3) apolipoprotein E-enriched lipoprotein A-I:A-II and lipoprotein A-I are the main apolipoprotein A-containing lipoproteins secreted by HepG2 cells. PMID- 2550001 TI - Immobilized GABAA receptors and their ligand binding characteristics. AB - ConA-sepharose and polylysine-agarose beads effectively bound detergent solubilized GABAA receptors from rat cerebrocortical membranes. The immobilized receptors showed a single class of high affinity binding sites specific for flunitrazepam or muscimol and displayed GABA-stimulated flunitrazepam binding. Maximal binding capacities of the ConA-immobilized receptor for the ligands were about three times greater than those of the polylysine-immobilized receptors. The relative affinities for each of the ligands were not affected by the method of receptor immobilization. The dissociation constants for muscimol of these immobilized receptors were somewhat dependent on the solubilizing agents used, but were considerably lower than those measured using extensively dialyzed rat cerebrocortical membranes. PMID- 2550002 TI - Chromium (V) and hydroxyl radical formation during the glutathione reductase catalyzed reduction of chromium (VI). AB - Electron spin resonance measurements provide evidence for the formation of long lived Cr(V) intermediates in the reduction of Cr(VI) by glutathione reductase in the presence of NADPH and for the hydroxyl radical formation during the glutathione reductase catalyzed reduction of Cr(VI). Hydrogen peroxide suppresses Cr(V) and enhances the formation of hydroxyl radicals. Thus Cr(V) intermediates catalyze generation of hydroxyl radicals from hydrogen peroxide through a Fenton like reaction. Thus the mechanism of Cr(VI) toxicity might involve the interaction between macromolecules and the hydroxyl radicals. PMID- 2550000 TI - Cholecystokinin elevates cytosolic calcium in small cell lung cancer cells. AB - The ability of cholecystokinin (CCK) to elevate intracellular Ca2+ levels in small cell lung cancer cells was investigated using the fluorescent Ca2+ indicator Fura 2. CCK-8 elevated the cytosolic Ca2+ levels in cell line NCI-H345 in a dose dependent manner. Nanomolar concentration of CCK-8 elevated cytosolic Ca2+ levels in the absence or presence of extracellular Ca2+. Potent CCK agonists such as gastrin-1 and nonsulfated CCK-8 but not inactive compounds such as CCK-27 32-NH2 elevated cytosolic Ca2+ levels. These data suggest that CCK receptors may regulate the release of Ca2+ from intracellular organelles in small cell lung cancer cells. PMID- 2550003 TI - Beta-endorphin receptors on cultured and freshly isolated lymphocytes from normal subjects. AB - [125I]-[D-Ala2]-beta EP*) binding to Epstein-Barr Virus (EBV) transformed B lymphocytes or to freshly isolated lymphocytes was characterized. The binding was time-, temperature- and pH-dependent; furthermore, it was reversible and cell concentration dependent. Maximum binding appeared at 15C in Tris buffer (pH 7.6) containing both BSA 0.5% and Bacitracin 0.1 mg/ml. beta EP inhibited BEP* binding to transformed lymphocytes and to freshly isolated lymphocytes by 50% at approximately 4 x 10(-8) M and 8 x 10(-9) M, respectively. Peptides representing amino-acid sequences 1-5, 1-16 and 1-17 of beta EP did not inhibit beta EP binding, neither did the opiates compounds Naloxone, Morphine, Bremazocine and Ethylketocyclazocine. On the contrary, beta EPd (6-31) inhibited beta EP* binding as effectively as beta EP, thus indicating that beta EP* binding to lymphocytes does not involve the N-terminal region of beta EP. 2 x 10(7)/ml freshly isolated lymphocytes bound 1.53 +/- 75%, whereas 2 x 10(6)/ml transformed lymphocytes bound 1.64 +/- 0.75% (mean +/- SD) beta EP* at tracer concentration. EBV transformed lymphocytes and freshly isolated lymphocytes may be useful to study beta EP receptors in humans. PMID- 2550004 TI - Modulation of benthiocarb in vitro inhibited neonate rat (Wistar strain) brain Na+K+-ATPase by norepinephrine. AB - Effect in vitro of benthiocarb, an organocarbamate herbicide on neonate rat (3 day old) brain was studied to understand the interaction of benthiocarb with Na+K+-ATPase. Na+K+-ATPase of the developing rat brain was selected as an index enzyme since alterations in the Na+K+-ATPase activity leads to neuronal dysfunction. The assay of Na+K+-ATPase in the presence of 1-8 mu moles of benthiocarb showed decreased activity and a concentration dependent inhibition of Na+K+-ATPase was noticed upto 7 mu moles of benthiocarb. Based on IC50 values (median concentration), 50% inhibition of the enzyme was observed with 5 mu moles of benthiocarb. Norepinephrine (NE) was selected to study the modulation of benthiocarb inhibited enzyme. Maximum increase (76.7%) of Na+K+-ATPase was noticed with 35 mu moles of NE and effective concentration (EC50) of NE which produced 50% activation of the enzyme was found to be 20 mu moles. This study suggests that NE acts as a protective agent in reversing the benthiocarb in vitro inhibited neonate rat brain Na+K+-ATPase. PMID- 2550005 TI - Serum protein degradation by hypochlorite. AB - The structural integrity of serum proteins: albumin, immunoglobulin G, transferrin, ceruloplasmin and superoxide dismutase, and the functional activity of the latter two enzymes after their interaction with hypochlorite were studied. It was shown that the interaction between the proteins and hypochlorite resulted in protein injury and degradation of their native structure. In the case of ceruloplasmin and transferrin, a practically complete protein "dissipation" occurred, the albumin and superoxide dismutase structures being injured in a lesser degree. The inactivation of ceruloplasmin was slower than that of superoxide dismutase. The protein degradation by hypochlorite seems to be the main factor restricting the ability of the proteins to act as antiinflammatory drugs. PMID- 2550007 TI - Evidence for a phosphatidylinositol anchor in glycolipid antigens of Leishmania major. AB - The structure of the membrane anchor of three Leishmania major surface antigenic glycolipids was analyzed. Phosphatidylinositol-specific phospholipase C treatment and nitrous acid deamination indicated the presence of a phosphatidylinositol anchor linked to the glycan through a non-N-acetylated hexosamine. An ester linkage on the C-2 of glycerol was revealed by phospholipase A2 hydrolysis. This fatty acyl substitution was not found on the phosphatidylinositol anchor of the Leishmania lipophosphoglycan. PMID- 2550006 TI - Effect of patulin on some enzymes of carbohydrate metabolism studied in rats. AB - The toxic nature of the secondary metabolite of Penicillium patulum has been studied in rats. Liver, Kidney and Intestine of the experimental animals showed derangement in carbohydrate metabolism. Changes in the concentration of a few key enzymes in carbohydrate metabolism have also been studied. Glycogen phosphorylase is found to be markedly increased while the glycolytic enzymes like hexokinase and aldolase are significantly lowered. Gluconeogenesis is stimulated and this is evidenced by increased glucose-6-phosphatase and fructose-1,6-diphosphatase activity. Our results revealed that, patulin, the secondary metabolite of Penicillium patulum showed toxicity in all the organs studied. PMID- 2550008 TI - Use of N-ethylmaleimide to directly determine the proportion of beta-adrenergic receptors exhibiting high affinity for agonists on human mononuclear leukocyte membranes. AB - Following incubation with the beta-adrenergic agonist isoproterenol (ISO) and the alkylating agent N-ethylmaleimide (NEM), the number of beta-adrenergic receptors measured on human mononuclear leukocyte (MNL) membranes decreased. Pretreatment induced an approximately 40% loss of measurable receptors. The proportion of receptors lost was that which formed the ternary complex of agonist-receptor-Gs protein (H-R-Gs) and exhibited high affinity for agonists. In comparison studies, the loss of receptors was found to be similar to the proportion of receptors that form the high affinity state of the receptor (%RH) for agonist in agonist competition binding studies. Acute exposure to agonist induced a similar reduction in ISO/NEM "sensitive" receptors and the %RH. These experiments indicate that pretreatment of MNL membranes with ISO/NEM is a powerful tool with which to directly determine the percentage of receptors able to form an agonist high affinity state. PMID- 2550009 TI - Inhibition of receptor-coupled phosphoinositide hydrolysis by sulfur-containing amino acids in rat brain slices. AB - Sulfur-containing amino acids were found to inhibit norepinephrine-stimulated [3H]phosphoinositide hydrolysis in rat cortical slices. Of the amino acids tested, L-cysteine was the most potent, inhibiting the response by 42 and 85% at concentrations of 50 and 500 microM respectively. L-Cystine and L-serine-O sulfate also inhibited the response to norepinephrine, but to a lesser degree than did L-cysteine. L-Homocysteic acid slightly potentiated phosphoinositide hydrolysis at a concentration of 100 microM, but caused inhibition at 500 microM. L-Cysteine sulfinate produced effects intermediate to those of L-cysteine and L homocysteic acid, having no effect on the response to norepinephrine at 50 microM, but causing 84% inhibition at 500 microM. The D-isomers of cysteine and homocysteic acid were much less potent than were the L-isomers. Examination of the time course of the inhibition of norepinephrine-stimulated [3H]phosphoinositide hydrolysis by L-cysteine showed that it was inhibited almost completely after 15, 30, 45 and 60 min of incubation. L-Cysteine and L homocysteic acid caused similarly strong inhibitions of the production of [3H]inositol monophosphate, [3H]inositol bisphosphate and [3H]inositol trisphosphate. The hydrolysis of [3H]phosphoinositides stimulated by norepinephrine in slices from rat hippocampus and striatum were inhibited by L cysteine to an extent similar to that occurring in cortical slices. These results demonstrate that several sulfur-containing amino acids, some of which have been proposed to be endogenous excitatory amino acid neurotransmitters, effectively modulate the response to norepinephrine of the phosphoinositide second messenger system in rat brain. PMID- 2550010 TI - Effect of cannabidiol on cytochrome P-450 isozymes. AB - Cannabidiol (CBD) has been shown to inhibit mouse hepatic mixed-function oxidations of several drugs after acute treatment, whereas repetitive treatment resulted in the restoration of drug-metabolizing capabilities. We have found that acute CBD treatment modestly decreased cytochrome P-450 content but markedly decreased hexobarbital hydroxylase, erythromycin N-demethylase, and 6 beta testosterone hydroxylase activities. Repetitive CBD treatment, on the other hand, resulted in the restoration of cytochrome P-450 content as well as hexobarbital hydroxylase and erythromycin N-demethylase activities. However, after such repeated treatments a fresh dose of CBD can once again inactivate erythromycin N demethylase activity but not hexobarbital hydroxylase activity. The resistance of hexobarbital hydroxylase to re-inactivation by CBD was paralleled by stimulation of pentoxyresorufin O-dealkylase activity and the appearance of a 50 kD protein that was immunoreactive to an antibody raised against rat hepatic cytochrome P 450b. CBD metabolism in vitro by microsomes prepared from such CBD-"induced" animals, resulted in a pattern of metabolites different from that observed from comparable incubations with liver microsomes from either untreated or phenobarbital-treated animals. Thus, it appears that CBD initially inactivates at least one cytochrome P-450 isozyme, but after repetitive CBD treatment, an isozyme is induced that is resistant to further re-inactivation by CBD. This isozyme appears to be immunochemically similar to, but somewhat functionally distinct from, the isozyme induced by phenobarbital treatment in mice. PMID- 2550011 TI - Intracellular lipoproteins as carriers for 2,3,7,8-tetrachlorodibenzo-p-dioxin and benzo(a)pyrene in rat and mouse liver. AB - The possible role of hepatic lipoproteins as intracellular carriers in the transport of 2,3,7,8-tetrachlorodibenzo-p-dioxin and benzo(a)pyrene was assessed by in vitro and in vivo studies. Following administration of [3H]2,3,7,8 tetrachlorodibenzo-p-dioxin or unlabelled 2,3,7,8-tetrachlorodibenzofuran to C57 BL/6 mice or Sprague-Dawley rats these compounds were bound to lipoproteins which subsequently underwent rapid and pronounced degradative processing, possibly catalysed by lipoprotein lipase, to heavier entities. At the highest doses of xenobiotics administered, an almost complete disappearance of lipoprotein particles was observed. The in vitro incubation of [3H]2,3,7,8-tetrachlorodibenzo p-dioxin-lipoprotein and [3H]benzo(a)pyrene-lipoprotein complexes with separated Ah receptor and 4S protein, respectively, demonstrated that a passive transfer occurred; the latter was likely dependent on both the relative affinities of the ligands towards the different cellular binding components as well as on their quantitative binding capacity. Taken together, these findings support the idea of a carrier-role for lipoproteins in the intracellular transport of hydrophobic xenobiotics and it may be asked whether the widespread modulators of lipoprotein level such as fibrates or others affect drug transfer or action. PMID- 2550013 TI - Gastrointestinal hormone receptors on isolated smooth muscle cells from gastric antrum of the rabbit. AB - The regulation by gastrointestinal polypeptide hormones of contraction and relaxation of functionally isolated smooth muscle cells from gastric antrum of the rabbit has been investigated. Gastrin, cholecystokinin (CCK-8) and motilin induced a rapid contraction of isolated cells: significant response occurred within a 5-sec incubation with these peptides and maximal response (40% decrease in cell length) after 30 sec. A higher sensitivity of smooth muscle cells to gastrin and CCK-8 than to motilin stimulations was demonstrated (EC50 = 10 pM for both gastrin and CCK-8 and EC50 = 1 nM for motilin). The minimal gastrin fragment required to get full contraction was the C-terminal pentapeptide amide common to gastrin and CCK. Proglumide inhibited gastrin- or CCK-8- but not motilin-induced contractions with an IC50 of 50 microM. contraction induced by gastrin and motilin required normal levels of extracellular calcium, whereas that due to CCK 8 seemed to be independent of extracellular calcium. Vasoactive intestinal polypeptide (VIP) caused a relaxation of smooth muscle cells maximally contracted by carbachol or CCK-8 or gastrin (EC50 = 2.2 nM) with a parallel increase in intracellular cAMP content. PMID- 2550012 TI - Partial GABA agonist activity of SR 95531 on the binding of [35S]TBPS, [3H]DMCM and [3H]lormetazepam to rat brain membranes. AB - A recently developed series of pyridazinyl-GABA derivatives has been classified as GABA antagonists in electrophysiological, behavioral and biochemical experiments. These substances seemed superior to the classical GABA antagonist bicuculline because of their water-solubility, high potency and apparent selectivity for GABAA receptors. In the present study the most potent representative of this class, SR 95531 almost completely reversed the stimulatory or inhibitory effect of GABA on [3H]lormetazepam and [35S]TBPS binding, respectively. To a lesser extent, it antagonized the inhibition of [3H]DMCM binding by GABA. However, the interaction of SR 95531 with the GABA receptor seems to be of a complex nature since the compound enhanced the binding of [3H]lormetazepam by 28% at 37 degrees in the presence of 200 mM Cl-. Bicuculline inhibited [3H]lormetazepam binding under these conditions, presumably by antagonizing the effect of residual endogenous GABA. Similar to GABA and THIP, SR 95531 potently inhibited the binding of [3H]DMCM and [35S]TBPS, suggesting SR 95531 to be a partial agonist at the GABAA receptor. PMID- 2550014 TI - Interactions between paraquat and ferric complexes in the microsomal generation of oxygen radicals. AB - Transition metals may play a central role in the toxicity associated with paraquat. Studies were carried out to evaluate the interaction of paraquat with several ferric complexes in the promotion of oxygen radical generation by rat liver microsomes. In the absence of added iron, paraquat produced some increase in low level chemiluminescence by microsomes; there was a synergistic increase in light emission in the presence of paraquat plus ferric-ATP or ferric-citrate, but not paraquat plus either ferric-EDTA or ferric-diethylenetriamine pentaacetic acid (ferric-DETAPAC). Synergistic interactions could be observed at a paraquat concentration of 100 microM and a ferric-ATP concentration of 3 microM. In the absence or presence of paraquat, microsomal light emission was not affected by catalase or dimethyl sulfoxide (DMSO), indicating no significant role for hydroxyl radicals. Superoxide dismutase (SOD) did not affect chemiluminescence in the absence of paraquat but produced some inhibition in the presence of paraquat; this inhibition by SOD was most prominent in the absence of added iron and less pronounced in the presence of ferric-ATP or ferric-citrate. Although microsomal chemiluminescence is closely associated with lipid peroxidation, paraquat did not increase malondialdehyde production as reflected by production of thiobarbituric acid-reactive components. However, lipid peroxidation was sensitive to inhibition by SOD in the presence, but not in the absence, of paraquat, analogous to results with chemiluminescence. Paraquat synergistically increased microsomal hydroxyl radical production as measured by the production of ethylene from 2-keto-4 thiomethylbutyrate in the presence of ferric-EDTA or ferric-citrate. The interaction of paraquat with microsomes and ferric complexes resulted in an increase in oxygen radical generation. Various ferric complexes can increase the catalytic effectiveness of paraquat in promoting microsomal generation of oxygen radicals, although, depending on the reaction being investigated, the nature of the ferric complex is important. PMID- 2550015 TI - Nicotine antagonists: phosphoinositide turnover and receptor binding to determine muscarinic properties. PMID- 2550016 TI - Serendipity in drug discovery. The case of BZR ligands. PMID- 2550017 TI - N-benzhydryl-tryptamines and 1,1-diphenyl-tetrahydro-beta-carbolines: synthesis, X-ray crystal structure and benzodiazepine receptor affinity. AB - Several N-benzhydryl-tryptamines and 1,1-diphenyl-tetrahydro-beta-carbolines were synthetized and the requisites for their formation were established. Crystallographic and conformational analyses were carried out on selected compounds and the affinity for the central benzodiazepine receptors was measured. PMID- 2550018 TI - Seroprevalence of human parvovirus B19 infection in Sao Tome and Principe, Malawi and Mascarene Islands. AB - The prevalence of antibodies to human parvovirus B19 (anti-B19 IgG) in sera (n = 577) from Sao Tome and Principe, Malawi and Mascarene Islands (Mauritius and Rodriguez Islands) was determined by antibody capture ELISA. The B19 prevalence was 51.5% on Sao Tome and Principe, 58.4% in Malawi, and 55.0% on the Mauritius mainland and 2.2% on Rodriguez Island, indicating that B19 virus is highly prevalent not only in Europe, Japan and the Americas but also in the African region. Rodriguez Islands has a very low B19 prevalence. PMID- 2550019 TI - Frequency of VH-gene utilization in human EBV-transformed B-cell lines: the most JH-proximal VH segment encodes autoantibodies. AB - We have studied VH-gene utilization in a collection of 187 IgM-secreting EBV transformed cell lines and have begun to correlate VH-gene family expression with binding properties of the secreted immunoglobulins. The results of these studies demonstrate that (1) frequency of VH-gene utilization in fetal and adult tissue derived cell lines correlates with the complexity of the family and (2) the single-membered most JH-proximal VH-6 family encodes autoantibodies reminiscent of autoantibodies found in the sera of patients with systemic lupus erythematosus. Nucleotide sequence analysis of VH-6-expressing clones revealed that each clone utilizes a short DH segment, resulting in a CDR3 region of conserved length. Our data suggest that EBV does not selectively transform human B cells on the basis of VH-gene family expression and that the VH-6 family encodes polyspecific autoantibodies that may serve an important regulatory function in the immune system. PMID- 2550020 TI - Two distinct mechanisms of interleukin-2 gene expression in human T lymphocytes. AB - Interleukin-2 (IL-2) gene regulation was investigated in primary cultures of highly purified human peripheral blood CD28+ T cells. Two discrete mechanisms for induction of T-cell proliferation could be distinguished by examining cell cycle progression and the expression of the IL-2 gene. Stimulation of cells by CD3 MoAb induced only transiently expressed, small amounts of IL-2 mRNA that was completely suppressed by cyclosporine. Costimulation of T cells with CD3 MoAb and either CD28 MoAb or PMA, but not calcium ionophore, induced a 50-100-fold increased in IL-2 gene expression and secretion. High levels of IL-2 gene expression could also be achieved by stimulation with calcium ionophore and PMA or CD28 MoAb and PMA, but not by CD28 MoAb plus calcium ionophore. IL-2 gene expression and T-cell proliferation induced by CD3 MoAb plus PMA or calcium ionophore plus PMA were completely suppressible by cyclosporine. In contrast, IL 2 gene expression and T-cell proliferation induced by CD28 MoAb plus PMA were unaffected by cyclosporine. The CD28 signal was dependent on new protein synthesis. Nuclear run-on transcription assays showed that anti-CD28 did not affect lymphokine transcription. A major effect of CD28 stimulation on mRNA stability was shown by studies using actinomycin D; CD28 stimulation substantially increased the half-life of IL-2 and TNF-alpha mRNA. The effects of anti-CD28 stimulation were specific for growth factors, and thus differ from previously described effects of cycloheximide on mRNA stability. These studies suggest the existence of two biochemical pathways for the induction of IL-2 production, one that occurs at the transcriptional level and is mediated by intracellular calcium release and protein kinase C and is cyclosporine-sensitive, and one that acts post-transcriptionally, is mediated by CD28 stimulation, and is cyclosporine-resistant. PMID- 2550021 TI - Characterization of anti-idiotypic antibodies generated against foot-and-mouth disease virus neutralizing monoclonal antibodies. AB - A series of seven neutralizing monoclonal antibodies (nMAbs) against type A12 foot-and-mouth disease virus (FMDV) was used to induce polyclonal anti-idiotypic antibodies (anti-ids) in rabbits. The anti-ids were semi-purified through isotype affinity columns and assayed by solid-phase radioimmunoassay for cross reactivity. nMAbs which map to the same epitope on the virion appear to contain a common idiotype, and the corresponding anti-ids competitively inhibited the virus nMAb reaction. Using a modified ELISA assay, it was possible to demonstrate binding of purified anti-ids to FMDV susceptible tissue culture cells. Such antibodies however, did not interfere with the binding of virus to cells, and the binding of anti-ids to FMDV receptor-negative cells could also be demonstrated. Mice were inoculated with purified anti-ids, and two elicited anti-viral antibodies, although these antibodies were non-neutralizing. Thus anti-ids to anti-FMDV nMAbs failed to react with cellular receptors for the virus, but were able to induce anti-viral antibody and thus should be examined as an alternative vaccine strategy for this virus. PMID- 2550022 TI - Studies on rotavirus homologous and heterologous active immunity in infant mice. AB - Homologous and heterologous active immunity was studied in mice with mammalian group A rotaviruses. One day old mice were vaccinated with one of the following rotaviruses: bovine B641 (serotype 6), bovine B223 (untyped), simian SA11 (serotype 3) and murine EDIM (untyped). At 10 days of age they were challenged with EDIM virulent virus or SA11 virus. All the vaccines induced a serological antibody response in the mice but only the homologous immune response was protective. PMID- 2550024 TI - Another encounter with an old foe: outbreak of poliomyelitis in Israel--1988. PMID- 2550023 TI - Induction of neutralizing antibodies to transmissible gastroenteritis virus by anti-idiotypic antibodies. AB - The potential of anti-idiotypic antibodies (anti-ids) as immunogens against transmissible gastroenteritis virus (TGEV) was tested in a heterologous system. A month-old pig was immunized with a neutralizing murine monoclonal antibody (MAb, 5A5) of the IgG2a isotype, specific for the E2 glycoprotein of TGEV. The anti-ids were isolated from the serum of the immunized pig by affinity chromatography, initially on a 5A5-Sepharose column, followed by repeated adsorption on a mouse IgG2a column. The swine anti-ids thus obtained bound to the MAb 5A5 (the idiotype), but not to MAbs of the same isotype IgG2a but of different idiotypes. The anti-ids also inhibited the binding of 5A5 to TGEV in a concentration dependent manner. Mice immunized with the anti-ids produced antibodies to TGEV. These antibodies, neutralized TGEV in vitro and inhibited the binding of 5A5 to TGEV. PMID- 2550025 TI - Inhibitory effects of alkane sulphonates on the function of immature rat Leydig, Sertoli and peritubular cells cultured in vitro. AB - Ethane 1,2-dimethane sulphonate (EDS) selectively destroys Leydig cells in the interstitium of the testis of adult rats. The toxic activity of this compound is much less obvious in the immature rat testis. We examined the effects of EDS, its monomethyl derivative and busulphan on cultured interstitial cells, percoll purified Leydig cells, Sertoli cells and peritubular cells derived from immature rats. The studies with interstitial cells and Leydig cells showed that EDS (40 160 micrograms/ml) blocked the conversion of C21 and androgen precursors into testosterone and androstenedione. Higher concentrations of this compound also inhibited the production of C21 steroids and the LH-induced production of cyclic AMP (cAMP). The observed effects required a latent period of at least 8 h and were slowly reversible. Isolated cells were more sensitive to EDS than monolayer cultures. Reaggregation cultures were even less sensitive. EDS was markedly more effective on immature Leydig cells than its monomethyl derivative and busulphan. In cultured Sertoli cells FSH-inducible aromatase activity, cAMP production, androgen-binding protein (ABP) production and the secretion of a paracrine factor with Leydig cell-stimulatory activity were markedly reduced by busulphan. In these cells, busulphan was clearly more active than EDS and its monomethyl derivative. The production of paracrine factors which increase ABP production and decrease FSH-inducible aromatase activity in Sertoli cells was studied as a parameter of the effects of alkane sulphonates on peritubular cells. Only busulphan markedly decreased the production of these paracrine factors. It is concluded that EDS displays a selective toxicity to Leydig cells derived from immature animals and that, apart from its effects on germ cells, busulphan may also directly impair the function of Sertoli cells and peritubular cells. PMID- 2550026 TI - Characterization of the receptor for glucagon-like peptide-1(7-36)amide on plasma membranes from rat insulinoma-derived cells by covalent cross-linking. AB - 125I-Labelled glucagon-like peptide-1(7-36)amide was cross-linked to a specific binding protein in plasma membranes prepared from RINm5F rat insulinoma-derived cells using disuccinimidyl suberate. Consistent with the presence of a single class of binding site on the surface of intact cells, only a single radiolabelled band at Mr63,000 was identified by SDS-PAGE after solubilization of the ligand binding protein complex. The band was not observed when 10nM glucagon-like peptide-1(7-36)amide was included in the binding assay, but 1 microM concentrations of glucagon-like peptide-1(1-36)amide, glucagon-like peptide-2 and glucagon did not decrease the intensity of labelling. No change in the mobility of the band was observed under reducing conditions, suggesting that the binding protein in the receptor is not attached to other subunits via disulphide bonds. In control incubations using plasma membranes from pig intestinal epithelial cells, which do not contain specific binding sites for glucagon-like peptide-1(7 36)amide, no cross-linked ligand-binding protein complex was observed. PMID- 2550027 TI - Effects of botulinum neurotoxin and Lambert-Eaton myasthenic syndrome IgG at mouse nerve terminals. AB - The interaction between two presynaptically acting agents, Lambert-Eaton myasthenic syndrome (LEMS) immunoglobulin G (IgG) and purified botulinum neurotoxin (BoNT) type A, was studied. Intracellular microelectrode recordings were carried out on mouse muscles after injection with LEMS IgG. BoNT was either injected before recordings were made or applied in vitro. The time course of the in vitro actions of BoNT on miniature end-plate potential and end-plate potential parameters were not affected by pretreatment with LEMS IgG. After in vivo injection of BoNT, end-plate potential quantal content was reduced to less than 2% of control values, whether or not LEMS IgG had also been previously given. Quantitative electron-microscope autoradiographical analysis showed that neither the binding of 125I-BoNT to acceptors on the nerve terminal membrane nor the pattern of its internalisation were affected by pretreatment with LEMS IgG. We conclude that the effects of BoNT are not affected by LEMS IgG, suggesting different presynaptic binding sites for the two agents. PMID- 2550028 TI - [Studies on the anti-virus effect of Glycyrrhiza uralensis Fish. polysaccharide]. AB - The anti-virus activity of GPS has been investigated in vitro using L-929 and FL cell line infected with 7 kinds of DNA or RNA virus, and the mechanism by which GPS exhibits the anti-virus activity has been studied. The results show that GPS may inhibit the growth of VSV, AdVIII, HSV-1 or VV and CPE is also suppressed by GPS for protecting culture cell from virus infection. PMID- 2550029 TI - Hemodynamic responses to converting enzyme inhibition in patients with renal disease. AB - We studied the effects of lisinopril on mean arterial blood pressure (MAP), plasma renin activity (PRA), and renal hemodynamics in nine patients with chronic renal disease and hypertension, before, and after three months of therapy. Lisinopril normalized blood pressure in five of nine patients (responders) and did not in the remaining four (nonresponders). PRA rose after lisinopril (4.8 +/- 2.6 ng/mL/h to 25 +/- 15 ng/mL/h, P less than 0.05) in responders, but not in nonresponders (2.0 +/- 1.4 ng/mL/h to 3.4 +/- 2.9 ng/mL/h). Glomerular filtration rate remained stable in both groups (responders: 43 +/- 11 mL/min to 43 +/- 22 mL/min; nonresponders: 39 +/- 25 mL/min to 32 +/- 21 mL/min). In the responders renal hemodynamics remained stable after lisinopril (renal plasma flow: 223 +/- 80 mL/min to 216 +/- 91 mL/min; filtration fraction: .20 +/- .04 to .20 +/- .05; renal vascular resistance: 386 +/- 179 to 326 +/- 209 units). In the nonresponders, renal plasma flow decreased (228 +/- 141 mL/min to 162 +/- 117 mL/min, P less than 0.005), filtration fraction increased (.19 +/- .08 to .24 +/- .12, P less than 0.05), and renal vascular resistance increased (695 +/- 747 to 1265 +/- 1574 units, P less than 0.05) after chronic lisinopril therapy. We conclude (1) there is a heterogeneous response to lisinopril in patients with chronic renal disease and hypertension, (2) lisinopril monotherapy may result in effective blood pressure control without renal hemodynamic compromise, and (3) an increase in PRA following converting enzyme inhibition may identify those in whom the circulating renin angiotensin system is participating in systemic hypertension and intrarenal hemodynamic changes. PMID- 2550030 TI - Erythrocyte sodium concentration and 86Rb uptake in weanling Dahl rats. AB - Alterations in Na, K ATPase pump activity as well as erythrocyte (RBC) intracellular sodium concentration (Nai) have been demonstrated in humans and rats with established hypertension. The contribution of hypertension itself to these changes is unclear. Accordingly, we investigated RBC ion transport and plasma ouabain-like factor (OLF) in four- to five-week old normotensive Dahl salt sensitive (DS) and salt-resistant (DR) rats on low salt diet. Although both strains were normotensive, systolic blood pressure (SBP) of DS (123 +/- 2 mm Hg) was higher than that of DR (116 +/- 1 mm Hg). No interstrain difference was evident in RBC pump activity measured as ouabain-sensitive 86rubidium (86Rb) uptake (DS = 0.277 +/- .030 and DR = 0.271 +/- .029 mumol/10(9)RBC/h) even though RBC Nai was greater in DS than DR (14.9 +/- 2.0 v 10.7 +/- 1.0 mEq/L; P less than 0.05). Plasma OLF was higher in DS than DR (28.9 +/- 4.7 v 16.5 +/- 2.3 pmol/mL; P less than 0.05), but did not correlate with RBC pump activity in either strain. RBC Nai was directly correlated with pump activity in DS (r = 0.84, P less than 0.01) and demonstrated a trend to correlate in DR (r = 0.71, P = 0.07). RBC Nai was also directly correlated with SBP in DR (r = 0.73, P less than 0.05) and DS (r = 0.70, P = 0.05). We conclude that RBC Nai is genetically determined in Dahl rats and is elevated in normotensive DS who are at risk for hypertension development.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550031 TI - Antihypertensive properties of linoleic acid and fish oil omega-3 fatty acids independent of the prostaglandin system. AB - Polyunsaturated fatty acids of the omega-6 and the omega-3 series have been shown to lower arterial pressure in humans and in various models of experimental hypertension by uncharacterized mechanisms. The objectives of our study were to compare the antihypertensive properties of linoleic acid (omega-6 series) and of fish oil fatty acids (omega-3 series) in a model of hypertension induced by the continuous subcutaneous infusion of angiotensin II in the rat and to determine whether or not their antihypertensive effects were mediated by the biosynthesis of vasodilator prostaglandins of classes 2 or 3. Linoleic acid and fish oil fatty acids (administered by subcutaneous injections) were equally potent in reducing, by half, the rise in systolic arterial pressure induced by the chronic infusion of angiotensin II. These antihypertensive effects were observed in the absence of any significant influence of either linoleic acid or fish oil fatty acids on the systemic and the renal synthesis of PGI2 or on the renal formation of PGE2 in vivo. Indomethacin caused a profound inhibition of the biosynthesis of PGI2 but not of PGE2 and could only partially neutralize the antihypertensive effects of linoleic acid and of fish oil fatty acids. These results suggest that, in this model of angiotensin II-induced hypertension, linoleic acid and fish oil fatty acids exert equipotent antihypertensive effects which are mainly independent of the prostaglandin system. PMID- 2550032 TI - POMC biosynthesis in the intermediate lobe of the spontaneously hypertensive rat. AB - Pro-opiomelanocortin (POMC) synthesized in the anterior (AL) and intermediate lobe (IL) of the rat pituitary gland, is a prohormone precursor of several peptide hormones that may participate in the regulation of blood pressure. We therefore studied the biosynthesis of POMC by measuring the incorporation of 3H tyrosine into the IL of 32-week-old spontaneously hypertensive rat (SHR) of the Aoki-Okamoto strain and their normotensive controls, the Wistar-Kyoto rat (WKY). Under basal conditions, POMC biosynthesis was significantly reduced in the SHR (1130 +/- 102 cpm/30 min/IL, n = 13) when compared to the WKY (1515 +/- 163 cpm/30 min/IL, N = 12, P less than 0.05, t test). There was also an inverse correlation between systolic blood pressure and POMC biosynthesis in both the WKY (Y = 8.4 +/- 1.38X + 2097 +/- 673, r = 0.86 +/- 0.06, N = 5), and SHR (Y = 5.7 +/ 1.8X + 1122 +/- 336 r = 0.57 +/- 0.13, N = 3). The decreased POMC biosynthesis was associated with a higher dopamine content in IL in SHR compared to WKY (1.56 +/- 0.53, n = 17 v 0.51 +/- 0.16, n = 17 pmol/IL, P less than 0.05, Mann-Whitney test). Oral administration of three different antihypertensive agents (propranolol, captopril, or hydralazine) for 12 weeks normalized blood pressure and POMC biosynthesis in the SHR but had no effect on either variable in the WKY. Because POMC biosynthesis in IL increased with normalization of blood pressure in the SHR, the decreased POMC biosynthesis in SHR may be a consequence rather than a cause of the elevated blood pressure in SHR. PMID- 2550033 TI - Cellular calcium regulation in hypertension. AB - In vascular muscle cells, two distinct types of functionally important calcium (Ca2+) channels, called transient (T) and sustained (L), are differentiated by dihydropyridine calcium antagonists (CaA). We studied the ratio of T/L Ca2+ channels in isolated, spontaneously contracting azygous venous cells of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) by quantitating Ca2+ currents and intracellular Ca2+ release. While total transmembranous Ca2+ current was not different between the two strains, the proportion of Ca2+ currents carried by L-type channels was enhanced in vascular muscle cells from SHR. We have recently compared subcellular distribution of intracellular free Ca2+ concentration in the same cells, at rest and during stimulation, by quantitation with a digital photon-counting camera. Fura-2 fluorescence intensity showed that Ca2+ release was principally from sarcoplasmic reticulum and that cells from SHR had higher levels of Ca2+ upon calcium channel stimulation, especially at the cell periphery. These findings suggest fundamental differences in SHR and WKY vascular muscle cells implicating the importance of changes in calcium channels, modulation of Ca2+ release, and Ca2+ uptake in SHR hypertension. PMID- 2550034 TI - Cyanide binding to canine myeloperoxidase. AB - Equilibria and kinetics of cyanide binding to canine myeloperoxidase were studied. Spectral results support the presence of two heme binding sites; an isosbestic point at 444 nm and a linear Scatchard plot suggest that the binding affinity of cyanide to the two subunits of the enzyme is the same. The dissociation constant is 0.53 microM. The pH dependence of the apparent second order rate constant indicates the presence of an acid-base group on the enzyme with a pKa of 3.8 +/- 0.1. The protonated form of cyanide binds to the basic enzyme with a rate constant of (4.3 +/- 0.3) x 10(6) M-1 s-1. PMID- 2550035 TI - Solubilization, characterization, and detergent interactions of lymphocyte 5' nucleotidase. AB - 5'-Nucleotidase is a member of a recently identified class of membrane proteins that is anchored via a phosphatidylinositol-containing glycolipid. The enzyme was readily solubilized with full retention of catalytic activity by nonionic and anionic detergents such as alkylthioglucosides, deoxycholate, and 3-[(3 cholamidopropyl)-dimethylammonio]-1-propane-sulfonate (CHAPS), while the cationic detergent dodecyltrimethylammonium bromide (DTAB) caused loss of activity. 5' Nucleotidase was released only at high detergent concentrations, suggesting that it is tightly associated with the membrane. DTAB and deoxycholate caused a loss of heat stability, while alkylthioglucosides had no effect. CHAPS produced a remarkable increase in the heat stability of the partially purified (glycoprotein fraction) and purified enzyme. Arrhenius plots of solubilized 5'-nucleotidase showed "break points" for all detergents in the temperature range 30-37 degrees C. SDS-PAGE of pure 5'-nucleotidase showed a single subunit of molecular mass 70 kilodaltons (kDa), while sucrose density gradient sedimentation gave a peak of activity corresponding to 132 kDa, indicating that the enzyme exists as a dimer. Gel filtration of the solubilized enzyme in several detergents showed apparent molecular masses between 200-630 kDa, suggesting that lymphocyte 5'-nucleotidase may be present in high molecular mass aggregates in its native state. PMID- 2550036 TI - The purification and properties of monoacylglycerol kinase from bovine brain. AB - Monoacylglycerol kinase (MGK) has been purified from bovine brain by six steps: isolation of cytosol, DEAE-cellulose chromatography, ammonium sulfate fractionation (0-40%), Bio-Gel A-1.5m, hydroxylapatite, and ATP-agarose column chromatography. The overall purification was 938 times with a 4.8% yield. The column separations (particularly Bio-Gel A-1.5m) and SDS- and nondenaturing polyacrylamide gel electrophoresis of enzyme purified from ATP-agarose indicated that MGK exists as a complex (approximately 350 kilodaltons) that is stabilized by 0.5 M NaCl and, on complete dissociation, yields a major protein of 72 kilodaltons. Dithiothreitol, EDTA, and ATP helped to stabilize MGK during purification. The protein peak eluted from hydroxylapatite by 25 mM phosphate activated and stabilized MGK activity. Phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin inhibited MGK. These phospholipids and others activated MGK synergistically with the above protein peak. MGK copurified with diacylglycerol kinase (DGK) throughout giving MGK to DGK ratios of 0.05-0.36. Optimal activity required 0.5 mM 2-monoolein and 10 mM MgCl2. Strong inhibition by p-chloromercuriphenyl sulfonic acid, N-ethyl-maleimide, and 5,5'-dithio-bis(2-nitrobenzoic acid), and prevention of this inhibition by dithiothreitol indicated the involvement of intact SH groups in the action of MGK. Purified MGK showed preference for substrates with unsaturated fatty acids except for 1- or 2-monostearin. Overall the preference favored the selective generation of 1-stearoyl- and 2-arachidonoyl-lysophosphatidic acid. PMID- 2550037 TI - [Changes in the levels of ACTH and cortisol after passive exposure to cigarette smoke in smokers and non-smokers]. AB - Plasma ACTH and cortisol levels were studied in smokers and non smokers, (exposed or not to smoke of the environment), after passive exposure to cigarette smoking. Non smokers, usually not exposed to smoke, show a rise in both hormones, whereas smokers and non smokers commonly exposed to smoke don't show any change in ACTH and cortisol levels. These data suggest that nicotine acts as an acute stimulus on the hypophysis-adrenal axis even passively inhaled. PMID- 2550038 TI - [Oral administration of nicotine and plasma levels of ACTH and cortisol in smokers and non-smokers]. AB - Plasma ACTH and cortisol levels after oral administration of nicotine (chewing gum containing nicotine 2 mg) in short and long time (10 and 45 min) were studied in smokers and non smokers. Non smokers after short time administration showed significant rise in ACTH and cortisol. No modifications were seen in the other groups of subjects. These data confirm that nicotine stimulates hypophysis adrenal axis in non smokers and that this effect is connected with nicotine contents of cigarettes rather than other volatile substances. PMID- 2550040 TI - [The culturing of human Schwann cells: a new contribution to the study of polyneuropathy]. AB - We have established 44 Schwann cell cultures from human peripheral nerves that underwent biopsy for diagnostic purposes using a new "sandwich" connective tissue and cut into 1 mm cubic explants which were placed between two glass coverslips coated with D-poly-L-lysine, in HAM-F10 medium, 15% FCS, 600 mg/dl glucose and antibiotics. In the first 3 weeks this new sandwich technique yields a fairly great and homogeneous amount of Schwann cell growth, with only a few fibroblasts and virtually no macrophages and provides a suitable substrate on which immuno cytochemical studies could be performed. PMID- 2550039 TI - [ACTH and cortisol after cigarette smoke exposure during the dexamethasone suppression test in smokers and non-smokers]. AB - In this research the effect of nicotine, (smoke of cigarette), was studied in smokers and non smokers during dexamethasone inhibitory test (1 mg h 23.00). ACTH and cortisol plasma levels, physiologically suppressed at 08.00, increased, after 30 min from smoking, only in group of non smokers. These data suggest that nicotine, in non smokers, could induce a maximum stimulus on diencephalic structures, so to overcome the inhibition of dexamethasone. PMID- 2550041 TI - The immature type of pro-opiomelanocortin cell of the rat anterior pituitary as observed by immunogold electron microscopy. AB - The perinatal development of glandular cells in the rat anterior pituitary producing pro-opiomelanocortin (POMC), the precursor protein of peptide hormones including adrenocorticotropin (ACTH), melanocyte-stimulating hormones (MSH) and endorphins was studied by an immunoelectron microscopic technique using the colloidal gold-antibody method. The POMC cells are classified into the following three types: 1) an immature type, 2) an intermediate type, and 3) a mature type, which correspond to Types III, I and II of the ACTH cells, respectively. The average size of the secretory granules in POMC cells varies widely, measuring 98.6 +/- 8.5 nm in the immature type, 111.4 +/- 6.7 nm in the intermediate type, and 139.3 +/- 23.1 nm in the mature type. The immature type comprises more than 90% of the POMC cells in the late fetal stage, but decreases in number after birth. The intermediate type reaches a peak at 8 days (female) or 33 days (male), while the mature type is that most frequently observed at 45 days, i.e., more than 50-60% of the total POMC cells, when the immature type decreases to about 15%. PMID- 2550042 TI - Experimental investigation of a new iodinated lipid emulsion for computed tomography of the liver. AB - Iodinated lipid emulsions are highly efficient macrophage imaging agents. Nevertheless, none of them has been accepted for clinical use because of adverse reactions. We have tested a new iodinated lipid emulsion, Intraiodol. The size and surface properties of the particles of this emulsion are similar to those of Intralipid which in turn closely resemble the naturally occurring chylomicrons. Using computed tomography (CT) of the rabbit liver as well as vital microscopy and electron microscopy of the rat liver we found that Intraiodol has low efficiency as a liver-specific contrast medium because its particles are predominantly taken up by the hepatocytes and to a less extent by the Kupffer cells, as is Intralipid. The low efficiency of Intraiodol could be fully compensated by an increase in dosage without any significant effect on sinusoidal blood flow. This in turn suggests that the likelihood of release of toxic mediators (and thereby related adverse reactions from activated macrophages) is reduced. We believe that this new way of delivering iodinated lipid particles to the liver represents an important advance in the search for a non-toxic lipid emulsion for CT of the liver. PMID- 2550043 TI - Transcatheter arterial chemotherapy using doxorubicin, iodized oil and Gelfoam embolization in hepatocellular carcinoma. AB - Computed tomography (CT) was performed in 54 patients with hepatocellular carcinoma three weeks after transcatheter arterial chemotherapy using iodized oil and doxorubicin with or without gelfoam embolization. Patients with iodized oil retention in the tumor greater than 50 per cent of tumor size survived longer than patients with retention of less than 50 per cent. Differences were also found within Okuda stages I and II, but they were significant only in Okuda stage I (p less than 0.0001). These results suggest a possible relationship between iodized oil retention and survival. In addition to Okuda stage, several factors affected iodized oil retention: tumor vascularity, tumor size, portal thrombosis and Gelfoam embolization. These factors may thus influence the prognosis after transcatheter arterial chemotherapy. PMID- 2550044 TI - In vivo microscopy of the liver after injection of Lipiodol into the hepatic artery and portal vein in the rat. AB - The route, distribution and clearance of intraarterially administered Lipiodol in the liver has been the subject of much speculation. The hepatic microcirculation was therefore studied by in vivo microscopy after injection of Lipiodol into the hepatic artery and the portal vein in rats. After intraarterial injection, Lipiodol rapidly entered the portal branches through arterio-portal communications. Lipiodol also passed through the sinusoids from the portal into the hepatic veins and then into the systemic circulation. Sinusoidal congestion occurred when the oil droplets filled the liver microcirculation and resolved as the oil was cleared. It is of clinical significance to note the passage of the oil into the systemic circulation after arterial injection. PMID- 2550045 TI - The pharmacokinetics and pharmacodynamics of perindopril in patients with hepatic cirrhosis. AB - 1. Perindopril, a new ACE inhibitor, is a prodrug requiring conversion into its active form perindoprilat by hydrolysis in the liver. 2. The pharmacodynamics and pharmacokinetics of perindopril (8 mg oral) and perindoprilat (2 mg intravenously) were studied in a double-blind randomised crossover study in a group of patients with compensated biopsy-proven hepatic cirrhosis. 3. Blood pressure and heart rate responses were similar after the two routes of administration as were plasma renin activity and aldosterone levels following dosing. 4. The AUC of perindoprilat after oral administration of perindopril represented 46 +/- 4% of the total AUC of perindopril and its metabolite when expressed in molar terms. Comparison with the AUC of perindoprilat after its intravenous administration suggested that 30 +/- 6% of the oral dose of perindopril was converted to its active metabolite. 5. The findings are comparable with those in healthy subjects. It appears that the presence of relatively mild hepatic cirrhosis does not significantly alter the pharmacokinetics of perindopril. PMID- 2550046 TI - Beta-adrenergic receptor responsiveness to isoprenaline in humans: concentration effect, as compared with dose-effect evaluation and influence of autonomic reflexes. AB - 1. Different techniques of assessing beta-adrenoceptor sensitivity in vivo, by use of i.v. infusions or bolus injections of isoprenaline (ISO), were compared in healthy volunteers. The importance of autonomic reflexes for responses to ISO was evaluated by studying the influence of 'autonomic blockade' by atropine and clonidine, which antagonize muscarinic effects and reduce sympathetic activity, respectively. Estimates of in vivo responsiveness to ISO were compared with parameters reflecting beta 2-adrenoceptor function in vitro in lymphocytes. 2. Heart rate responses to infused ISO were not significantly altered by 'autonomic blockade' when evaluated from concentration-effect curves. When related to the infused dose of ISO, however, sensitivity was artefactually increased (P less than 0.05), as the plasma concentrations of ISO were 40% higher after atropine and clonidine. Heart rate responses to bolus injections of ISO were attenuated (P less than 0.05) by 'autonomic blockade', suggesting that facilitatory reflexes contribute to these non-steady state responses. Intersubject variations in heart rate responsiveness to ISO were greater than the intrasubject variability caused by counterregulatory reflexes. 3. 'Autonomic blockade' lowered venous plasma noradrenaline at rest. The noradrenaline response to ISO infusion was attenuated and the diastolic blood pressure response enhanced, indicating that a counterregulatory vasoconstrictor reflex normally is activated by ISO-induced vasodilatation. The plasma cyclic AMP response to ISO, on the other hand, was unaffected by atropine and clonidine and reflects beta 2-adrenoceptor responsiveness in vivo. 4. In vitro data for beta-adrenoceptor binding sites (Bmax;[125I]-IHYP binding) and cyclic AMP responses to ISO in lymphocytes correlated with DBP and noradrenaline responses to infused ISO. No correlations were found between in vitro data and heart rate, plasma cyclic AMP or plasma glycerol responses to infused ISO in vivo. 5. During prolonged ISO infusions (in six other healthy subjects) physiological responses reached greater than 90% of their steady state level after 8 min, but no definite steady state level could be defined for the plasma concentration of ISO during 40 min of infusion. 6. The ISO infusion test showed a good reproducibility, especially when repeated on the same day. Evaluation of plasma concentration-effect relationships increase the precision of the ISO infusion test as confounding inter- and intra-individual variations in ISO concentrations (as caused by e.g. autonomic blockade) will be taken into account.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2550047 TI - Investigations on plasma activity of low molecular weight heparin after intravenous and oral administrations. AB - This study investigated the absorption of low molecular weight heparin (LMWH) after oral administration in man. Six healthy subjects received 5,000 anti-Xa units of LMWH (Kabi 2165) by both i.v. and oral administration. The oral formulations were prepared in pH 4.0 and pH 7.0 buffered solutions. Multiple blood samples were collected after each dose for measurements of anti-Xa, anti IIa and APTT plasma heparin activities. Pharmacokinetic parameters based on the anti-Xa activity measurements after the i.v. dose were as follows (mean +/- s.d.): t1/2, 1.82 +/- 0.23 h; V, 4.34 +/- 0.651; and CL, 28.0 +/- 6.2 ml min-1. Half-life values based on the anti-IIa and APTT activities were 1.63 +/- 0.43 and 1.09 +/- 0.51 h, respectively. Considerable prolongations in APTT were observed, with APTT at 30 min averaging 55.7 +/- 4.1 s (1.84 +/- 0.27 times baseline values). After oral administration, no measurable plasma heparin activities were observed with either LMWH preparation. The results of this investigation indicate that LMWH does not have detectable plasma activity after oral administration, and that after i.v. administration it has significant anti-IIa and APTT activities in addition to its anti-Xa activity. PMID- 2550049 TI - Relation between asbestosis and bronchial cancer in amphibole asbestos miners. AB - In a necropsy series of 339 amphibole asbestos miners heavy smoking, age, and the presence of asbestosis were significantly associated with the presence of bronchial cancer. Of the 35 cases of bronchial cancer, 24 were associated with asbestosis. Eleven cases of bronchial cancer occurred in men without asbestosis; all were smokers. Standardised proportional mortality rates indicated no excess of bronchial cancer in 302 exposed men without asbestosis whereas these rates were progressively raised in men with slight or moderate/severe asbestosis. Of the four exposure variables introduced separately into a logistic regression model, "years of exposure" made a small but significant contribution; "residence time" marginally failed to achieve a 5% level of significance. Two other exposure variables tested including cumulative fibre exposure (fibre years) made no significant contribution. In the absence of asbestosis at necropsy a bronchial cancer in a man exposed to asbestos is unlikely to be due to asbestos. PMID- 2550048 TI - Cancer mortality in relation to measures of occupational exposure to crocidolite at Wittenoom Gorge in Western Australia. AB - The separate and combined effects of duration and intensity of exposure to crocidolite on mortality from lung cancer, malignant mesothelioma, and stomach cancer were examined in 6506 male former crocidolite miners and millers at Wittenoom Gorge, Western Australia. Each subject who had died from lung cancer (92), mesothelioma (31), or stomach cancer (17) was matched with up to 20 control subjects of the same age who were not known to have died before the index subject. Relations of dose and time of exposure to crocidolite to risk of death were modelled by conditional logistic regression. For lung cancer, the best fitting multiplicative model was one which estimated a relative risk (RR) of 1.12 (95% CI 1.04-1.20) per year of exposure and 1.01 (95% CI 1.00-1.01) per fibre/ml. This was statistically indistinguishable from an additive model showing an increase in RR of 0.01045 (95% CI 0.008-0.020) per f/ml year. For mesothelioma the best fitting model appeared to be one estimating a RR of 24.9 (95% CI 3.51 1.77) per log year since first exposed and a RR of 10.5 (95% CI 3.12-35.1) if exposed for longer than six months. This was not distinguishable statistically from a model that showed mortality increasing as the fourth power of time since first exposed less the fourth power of time since last exposed. The effect of intensity of exposure on the RR for mesothelioma was only slight. There was no consistent effect of any measure of exposure to crocidolite on death from stomach cancer. PMID- 2550050 TI - Pump-1 cDNA codes for a protein with characteristics similar to those of classical collagenase family members. AB - Pump-1 cDNA has recently been isolated by screening a human tumor cDNA library with a transin (rat stromelysin) probe under low-stringency hybridization conditions. The cDNA codes for a potential protein with significant sequence similarity to the metalloproteinases collagenase and stromelysin, but which lacks the hemopexin-like domain characteristic of these enzymes. Expression of pump-1 cDNA in cos cells using an expression vector leads to secretion of a protein of Mr 28,000 with latent, organomercurial-activatable proteinase activity. Cos cells transfected with a partial pump-1 cDNA in the vector pPROTA secrete a fusion protein between the IgG-binding domains of staphylococcal protein A and pump-1. The fusion protein binds to IgG-Sepharose, and the bound fusion protein undergoes apparent autocleavage in the presence of 4-aminophenylmercuric acetate with elution of active pump-1 species of Mr 21,000 and 19,000. Active pump-1 degrades casein, gelatins of types I, III, IV, and V, and fibronectin and can activate collagenase. Active pump-1 is inhibited by EDTA, 1,10-phenanthroline, and the tissue inhibitor of metalloproteinases. These results show that, despite the absence of a hemopexin-like domain, pump-1 is a latent secreted metalloproteinase. Postpartum rat uteri contain elevated levels of rat pump-1 mRNA. On the basis of this observation, its size, and its substrate specificity, we suggest that pump-1 might correspond to a previously described uterine metalloproteinase, matrix metalloproteinase 7. PMID- 2550052 TI - Kinetic and structural effects of activation of bovine kidney aldose reductase. AB - Aldose reductase, purified to homogeneity from bovine kidney, is converted in a temperature-dependent process from a low-Km/low-Vmax form to a high-Km/high-Vmax form of the enzyme. Activation, which results in significant changes in the protein secondary structure, as detected by fluorescence spectroscopy, circular dichroism, and thiol modification with 5,5'-dithiobis(2-nitrobenzoic acid), has no effect on the apparent Mr, pI, or homogeneity of the enzyme, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and agarose isoelectric focusing. Vmax, which varied less than 3-fold for a series of aldehyde substrates with either activation state of the enzyme, increased an average of (17 +/- 4) fold upon activation of the enzyme. V/Kaldehyde increased or decreased up to 4 fold, depending on the substrate. Activation desensitized the enzyme to inhibition by aldose reductase inhibitors, with the apparent Ki value increasing from 2-fold for Epalrestat [ONO-2235, (E)-3-(carboxymethyl)-(E)-5-[2-methyl-3 phenylpropenylidene]-rhoda nine] to 200-fold for AL-1576 (spiro [2,7 difluorofluorene-9,4'-imidazolidine]-2',5'-dione). Biphasic double-reciprocal plots for the aldehyde substrates and biphasic Dixon plots for inhibition by AL 1576 and Statil [ICI-128,436; 3-[(4-bromo-2-fluorobenzyl)-4-oxo-3H-phthalazin-l ylacetic acid], observed during the course of activation, are quantitatively accounted for by the individual contributions of the two enzyme forms. On the basis of an analysis of the kinetic data, a mechanism is proposed in which isomerization of the free enzyme limits the rate of the forward reaction for the unactivated enzyme and is the primary step affected by activation. PMID- 2550051 TI - Characterization of genes encoding rat tonin and a kallikrein-like serine protease. AB - Tissue kallikreins are a group of serine proteases which may function as peptide hormone processing enzymes. Two rat kallikrein genomic clones (RSKG-5 and RSKG 50) were sequenced and characterized. The rat tonin gene and a kallikrein-like gene were found in clones RSKG-5 and RSKG-50, respectively. The tonin gene is 4146 base pairs in length, with both the variant CCAAA and TTTAAA boxes in the 5' end region and an AATAAA polyadenylation signal at the 3' end of the gene. It has five exons which are separated by four introns. Sequence analysis of 3.7-kb 5' upstream and 7.5-kb 3' downstream of the tonin gene failed to reveal a second kallikrein gene. Sequence comparisons of the RSKG-5 exons with tonin cDNA revealed that only one base in the 3'-noncoding region was different from that in the previously reported rat tonin cDNA. Characteristic TC- and TG-repeated sequences were also found in the first and second introns of the tonin gene. The tonin gene encodes a preprotonin of 259 amino acids (aa). The active enzyme consists of 235 aa and is preceded by a deduced signal peptide of 17 aa and a profragment of 7 aa. Northern blot analysis indicates that RSKG-5 is expressed in a sex-dependent manner in rat submandibular gland, with a higher level expressed in males. The RSKG-50 gene was truncated at an EcoRI site in the second intron, excluding its 5' end. Compared to the coding sequence of pancreatic kallikrein, 12 nucleotides have been deleted in exon 3 of the RSKG-50 gene. The nucleotide sequences of the third, fourth, and fifth exons of the RSKG-50 gene encode a polypeptide of 188 aa residues. The translated peptide is 80% homologous to rat pancreatic kallikrein and 75% homologous to rat tonin in the corresponding regions. Key residues in the RSKG-50 gene product indicate a serine protease with kallikrein-like cleavage specificity at basic amino acids. PMID- 2550053 TI - Coenzyme binding by 3-hydroxybutyrate dehydrogenase, a lipid-requiring enzyme: lecithin acts as an allosteric modulator to enhance the affinity for coenzyme. AB - The role of phospholipid in the binding of coenzyme, NAD(H), to 3-hydroxybutyrate dehydrogenase, a lipid-requiring membrane enzyme, has been studied with the ultrafiltration binding method, which we optimized to quantitate weak ligand binding (KD in the range 10-100 microM). 3-Hydroxybutyrate dehydrogenase has a specific requirement of phosphatidylcholine (PC) for optimal function and is a tetramer quantitated both for the apodehydrogenase, which is devoid of phospholipid, and for the enzyme reconstituted into phospholipid vesicles in either the presence or absence of PC. We find that (i) the stoichiometry for NADH and NAD binding is 0.5 mol/mol of enzyme monomer (2 mol/mol of tetramer); (ii) the dissociation constant for NADH binding is essentially the same for the enzyme reconstituted into the mixture of mitochondrial phospholipids (MPL) (KD = 15 +/- 3 microM) or into dioleoyl-PC (KD = 12 +/- 3 microM); (iii) the binding of NAD+ to the enzyme-MPL complex is more than an order of magnitude weaker than NADH binding (KD approximately 200 microM versus 15 microM) but can be enhanced by formation of a ternary complex with either 2-methylmalonate (apparent KD = 1.1 +/ 0.2 microM) or sulfite to form the NAD-SO3- adduct (KD = 0.5 +/- 0.1 microM); (iv) the binding stoichiometry for NADH is the same (0.5 mol/mol) for binary (NADH alone) and ternary complexes (NADH plus monomethyl malonate); (v) binding of NAD+ and NADH together totals 0.5 mol of NAD(H)/mol of enzyme monomer, i.e., two nucleotide binding sites per enzyme tetramer; and (vi) the binding of nucleotide to the enzyme reconstituted with phospholipid devoid of PC is weak, being detected only for the NAD+ plus 2-methylmalonate ternary complex (apparent KD approximately 50 microM or approximately 50-fold weaker binding than that for the same complex in the presence of PC). The binding of NADH by equilibrium dialysis or of spin-labeled analogues of NAD+ by EPR spectroscopy gave complementary results, indicating that the ultrafiltration studies approximated equilibrium conditions. In addition to specific binding of NAD(H) to 3 hydroxybutyrate dehydrogenase, we find significant binding of NAD(H) to phospholipid vesicles. An important new finding is that the nucleotide binding site is present in 3-hydroxybutyrate dehydrogenase in the absence of activating phospholipid since (a) NAD+, as the ternary complex with 2-methylmalonate, binds to the enzyme reconstituted with phospholipid devoid of PC and (b) the apodehydrogenase, devoid of phospholipid, binds NADH or NAD-SO3- weakly (half maximal binding at approximately 75 microM NAD-SO3- and somewhat weaker binding for NADH).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2550054 TI - Autophosphorylation of the type II calmodulin-dependent protein kinase is essential for formation of a proteolytic fragment with catalytic activity. Implications for long-term synaptic potentiation. AB - Autophosphorylation plays an essential role in proteolytic activation of the type II calmodulin-dependent protein kinase (CaM kinase II). Limited proteolysis of CaM kinase II by trypsin, alpha-chymotrypsin, and Ca2+-stimulated neutral protease (calpain) yielded a catalytically active kinase fragment only when the holoenzyme was autophosphorylated prior to proteolysis. Slightly larger, inactive fragments were obtained from nonphosphorylated CaM kinase II, regardless of whether Ca2+/calmodulin or Mg2+/ATP were present or absent. The active fragment exhibited Ca2+/calmodulin-dependent kinase activity with kinetic parameters identical with those of the activated holoenzyme. The key autophosphorylation site of CaM kinase II was absent from the active fragment which indicates that proteolysis can effectively uncouple the activation state and Ca2+/calmodulin independence of the kinase from the action of phosphoprotein phosphatases. Because autophosphorylation exerts such a tight control over this irreversible process, proteolytic activation of CaM kinase II by intracellular proteases offers an attractive mechanism for prolonging the effects of Ca2+ at the synapse. PMID- 2550055 TI - Characterization of four herbicide-resistant mutants of Rhodopseudomonas viridis by genetic analysis, electron paramagnetic resonance, and optical spectroscopy. AB - Herbicides of the triazine class block electron transfer in the photosynthetic reaction centers of purple bacteria and PSII of higher plants. They are thought to act by competing with one of the electron acceptors, the secondary quinone, QB, for its binding site. Several mutants of the purple bacterium Rhodopseudomonas viridis resistant to terbutryn [2-(methylthio)-4-(ethylamino)-6 (tert-butylamino)-s-triazine] have been isolated by their ability to grow photosynthetically in the presence of the herbicide. Sequence analysis of the genes coding for the L and M subunits of the reaction center showed that four different mutants were obtained, two of them being double mutated: T1 (SerL223--- Ala and ArgL217----His), T3 (PheL216----Ser and ValM263----Phe), T4 (TyrL222--- Phe), and T6 (PheL216----Ser). The residues L223 and L216 are involved in binding of QB, whereas L217 and L222 are not. M263 is part of the binding pocket of the primary quinone, QA. The affinity of the reaction centers for terbutryn and the electron transfer inhibitor o-phenanthroline, determined via the biphasic charge recombination after one flash, is decreased for all mutants. The affinity for ubiquinone 9 is also decreased, except in T1. Characterization by EPR spectroscopy showed that the QB.-Fe2+ signal of T4, having a g = 1.93 peak, is different from the signals obtained with the wild type and the other mutants but very similar to those of Rhodospirillum rubrum and PSII. The results obtained by the combination of these different techniques are discussed with respect to the three-dimensional structure of the wild type and the mode of binding of ubiquinone, terbutryn, and o-phenanthroline as determined by X-ray structure analysis. PMID- 2550056 TI - Topographical characterization of the domain structure of the bovine adrenal atrial natriuretic factor R1 receptor. AB - We have studied the structure and function of the membrane atrial natriuretic factor R1 (ANF-R1) receptor using limited proteolysis and exoglycosidase treatment. Limited digestion with trypsin of the receptor from bovine adrenal zona glomerulosa membranes resulted in the conversion of the native 130-kDa receptor into a single membrane-associated ANF-binding proteolytic fragment of 70 kDa. The 70-kDa fragment bound ANF with enhanced binding affinity but retained intact ANF-R1 pharmacological specificity and was still sensitive to modulation by amiloride. Trypsin treatment of the membranes produced a dual effect on ANF binding. Low concentrations of trypsin (less than or equal to 25 micrograms/mg of protein) increased ANF binding while higher concentrations dose dependently reduced the binding of the hormone. The increase of ANF-binding activity was associated with the formation of the 70-kDa fragment while the loss of ANF binding paralleled the degradation of the 70-kDa fragment. Low concentrations of trypsin drastically decreased the ANF-sensitive guanylate cyclase activity of the membrane fraction. This loss of catalytic activity strongly correlated with the formation of the 70-kDa tryptic fragment. We also evaluated the effect of ANF binding on the susceptibility of the receptor to proteolytic cleavage. The occupied receptor exhibited a greater sensitivity to trypsin digestion than the unoccupied protein, consistent with the hypothesis that hormone binding induces an important conformational change in the receptor structure. On the other hand, the 70-kDa fragment was much more resistant to proteolysis when occupied by ANF, suggesting that the ANF-binding domain forms a very compact structure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550057 TI - Rotational motion of yeast cytochrome oxidase in phosphatidylcholine complexes studied by saturation-transfer electron spin resonance. AB - Cytochrome oxidase from yeast has been covalently labeled with a nitroxide derivative of maleimide and reconstituted in lipid-substituted complexes with dimyristoyl-, dioleoyl-, or dielaidoyl-phosphatidylcholine. The rotational mobility of the enzyme in the complexes has been studied as a function of temperature and time, and of lipid/protein ratio, using saturation-transfer electron spin resonance spectroscopy. For complexes with dimyristoylphosphatidylcholine, the rotational mobility of the protein decreases abruptly below the gel-to-fluid-phase transition. This change is accompanied by a lateral segregation of the protein, as seen by freeze-fracture electron microscopy, and by an increase in the activation energy for the enzymatic activity. A time-dependent decrease in the rotational motion of the protein is observed on incubating at temperatures in the fluid phase of the lipid. This corresponds with a time-dependent loss of enzyme activity observed on incubation at temperatures in the fluid phase, but not at temperatures in the gel phase, over a period of 3 h. The rotational mobility decreases with increasing protein concentration in the complexes, both in the fluid and in the gel phases. The dependence of the protein mobility on lipid/protein ratio can be interpreted quantitatively in terms of the effect of increased random protein-protein contacts in the fluid phase. The maximum limiting rotational correlation time for the protein diffusion at high lipid/protein ratios in the fluid phase is tau R[[ approximately equal to 25 microseconds, suggesting that the protein is present as either a monomer or more probably a dimer in the reconstituted membrane. PMID- 2550058 TI - High mobility group protein 1 preferentially conserves torsion in negatively supercoiled DNA. AB - HMG 1 is known to bind to a variety of DNAs and to unwind nicked and closed circular DNA. We now report evidence that it has a significantly higher unwinding angle on negatively supercoiled DNA than on the other torsional forms. The degree of unwinding observed on nicked circular DNA depends on the purity of the HMG 1 preparation used. HMG 1 from CM-Sephadex has an unwinding angle of 28.8 degrees, compared to 7.2 degrees for the purer preparation obtained from Mono S, suggesting that contaminating strand-separating activity is removed by the additional purification step. The subsequent studies on closed circular forms of DNA were all performed using the purer HMG 1. After preincubation of highly negatively supercoiled DNA (sigma = -0.040) with HMG 1, the DNA-protein mixture was relaxed with Escherichia coli topoisomerase I. At molar ratios of less than 100:1 (HMG 1 to DNA), negatively supercoiled DNA displays a dose-dependent change in the linking number, indicating an unwinding angle of 57.6 degrees. HMG 1 protects 50% of highly negatively supercoiled DNA from E. coli topoisomerase I at a molar ratio of 100:1, and protects all supercoils at a molar ratio of 200:1, indicating saturation of the DNA at this concentration. HMG 1 also protects highly negatively supercoiled DNA from calf thymus topoisomerase I, with an apparent unwinding angle of 57.6 degrees. Moderately negatively supercoiled DNA (sigma = -0.018), but not moderately positively supercoiled DNA (sigma = +0.011), competes for the protective effect of HMG 1 on highly negatively supercoiled DNA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550059 TI - Purification and characterization of an altered topoisomerase II from a drug resistant Chinese hamster ovary cell line. AB - The cytotoxicity and DNA damage induced by the epipodophyllotoxins and several intercalating agents appear to be mediated by DNA topoisomerase II. We have purified topoisomerase II to homogeneity both from an epipodophyllotoxin resistant Chinese hamster ovary cell line, VpmR-5, and from the wild-type parental cell line. Immunoblots demonstrate similar topoisomerase II content in these two cell lines. The purified enzymes are dissimilar in that DNA cleavage by VpmR-5 topoisomerase II is not stimulated by VP-16 or m-AMSA. Furthermore, the VpmR-5 enzyme is unstable at 37 degrees C. Thus, the drug resistance of VpmR-5 cells appears to result from the presence of an altered topoisomerase II in these cells. Purified topoisomerase II from VPMR-5 and wild-type cells has the same monomeric molecular mass as well as equivalent catalytic activity with respect to decatenation of kinetoplast DNA. Etoposide (VP-16) inhibits the activity of both enzymes. Noncovalent DNA-enzyme complex formation, assayed by nitrocellulose filter binding, is also similar, as is protection from salt dissociation of this complex by ATP and VP-16. The data suggest a model in which the drug-resistant cell line, VpmR-5, has religation activity which is less affected by drug than that of the wild-type cells. Drug effect on DNA religation and catalytic activity are dissociated mechanistically. In addition, under certain circumstances, the "cleavable complex" observed following denaturation of a drug-stabilized DNA enzyme complex may not adequately reflect the nature of the antecedent lesion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550060 TI - Design of a selective insulin receptor tyrosine kinase inhibitor and its effect on glucose uptake and metabolism in intact cells. AB - An inhibitor of the insulin receptor tyrosine kinase (IRTK), (hydroxy-2 naphthalenyl-methyl) phosphonic acid, was designed and synthesized and was shown to be an inhibitor of the biological effects of insulin in vitro. With a wheat germ purified human placental insulin receptor preparation, this compound inhibited the insulin-stimulated autophosphorylation of the 95-kDa beta-subunit of the insulin receptor (IC50 = 200 microM). The ability of the kinase to phosphorylate an exogenous peptide substrate, angiotensin II, was also inhibited. Half-maximal inhibition of basal and insulin-stimulated human placental IRTK activity was found at concentrations of 150 and 100 microM, respectively, with 2 mM angiotensin II as the peptide substrate. The inhibitor was found to be specific for tyrosine kinases over serine kinases and noncompetitive with ATP. The inhibitor was converted into various (acyloxy)methyl prodrugs in order to achieve permeability through cell membranes. These prodrugs inhibited insulin stimulated autophosphorylation of the insulin receptor 95-kDa beta-subunit in intact CHO cells transfected with human insulin receptor. Inhibition of insulin stimulated glucose oxidation in isolated rat adipocytes and 2-deoxyglucose uptake into CHO cells was observed with these prodrugs. Our data provide additional evidence for the involvement of the insulin receptor tyrosine kinase in the regulation of glucose uptake and metabolism. These results and additional data reported herein suggest that this class of prodrugs and inhibitors will be useful for modulating the activity of a variety of tyrosine kinases. PMID- 2550061 TI - Electron spin echo envelope modulation studies of pyruvate kinase active-site complexes. AB - Electron spin echo envelope modulation (ESEEM) spectroscopy, with Mn2+ and VO2+ as paramagnetic probes, was used to examine active-site structures at the protein based divalent cation site of rabbit muscle pyruvate kinase in the presence of substrates, products, and the requisite inorganic cofactors. Two different VO.protein complexes were clearly distinguished, which differed with respect to coordination of the active-site lysine to VO2+. Lysine coordination was sensitive to the presence of pyruvate and phosphoenolpyruvate (PEP) and to the nature of the monovalent cation. In the presence of MgATP and oxalate, a 4-MHz 31P contact interaction was observed, which indicates that the ATP is directly coordinated to Mn2+ at the protein-based site. No 31P contact interactions were observed, however, in the presence of PEP. Pyruvate was determined to be a bidentate ligand of VO2+, on the basis of the observation of 2.2- and 5.4-MHz 13C contact interactions between VO2+ and [1-13C]pyruvate and [2-13C]pyruvate, respectively. Magnetic coupling between VO2+ or Mn2+ and 23Na, 39K, and 133Cs was observed, demonstrating the close proximity of the monovalent cation and the protein-based divalent cation. PMID- 2550062 TI - Thiol and amino analogues as alternate substrates for glycerokinase from Candida mycoderma. AB - The kinetic and catalytic mechanism of glycerokinase from Candida mycoderma was examined with thiol and amino analogues of glycerol and with MgAMPPCP, an analogue of MgATP. (S)-1-Aminopropanediol was phosphorylated on nitrogen (Vmax 0.4% that of glycerol) while the R enantiomer was phosphorylated on oxygen (Vmax 0.7% that of glycerol). (S)-1-Mercaptopropanediol was phosphorylated on oxygen (Vmax 3.5% that of glycerol), while the R enantiomer was phosphorylated on sulfur (Vmax 0.001% that of glycerol). The hydroxyl group at C-2 thus orients the substrate in the active site, while that at the carbon remote from phosphorylation enhances both catalysis and binding of the substrate, presumably because of hydrogen-bonding interactions. The kinetic mechanism is random with a high degree of synergistic binding between the substrates, so that the mechanism appears ordered with glycerol adding first but equilibrium ordered with MgATP binding first with the amino analogues. PMID- 2550063 TI - EPR and kinetic analysis of the interaction of halides and phosphate with nitrate reductase. AB - Electron paramagnetic resonance spectra obtained during turnover of the Mo center of NADH:nitrate reductase at pH 8 were comprised of two Mo(V) species, signal A (g1 = 1.996, g2 = 1.969, g3 = 1.967, A1H = 1.25 mT, A2H = 1.18 mT, and A3H = 1.63 mT) and signal B (g1 = 1.996, g2 = 1.969, and g3 = 1.967), the former exhibiting superhyperfine interaction due to strong coupling with a single, exchangeable proton. Binding of halides and nitrite to the Mo center increased the proportion of signal A whereas phosphate had no effect on the EPR line shape. Halides decreased and phosphate increased the rates of enzyme activities involving the Mo center (NADH:nitrate reductase and reduced methyl viologen:nitrate reductase), but neither had any effect on activities involving FAD (NADH:ferricyanide reductase) or heme (NADH:cytochrome c reductase), indicating specific binding of halides to the Mo center. Halides were found to be weak, mixed competitive noncompetitive inhibitors (Cl- KI = 39 mM, mu = 0.2 M, pH 8) of nitrate reductase forming a catalytically inactive ternary halide-nitrate-enzyme complex. Inhibition patterns changed from nearly noncompetitive (F-) to nearly competitive (I-). The weakening of nitrate binding due to halide binding correlated with increased halide electronegativity rather than ionic radius. In contrast, phosphate (Kd = 7.4 mM, mu = 0.2 M, pH 8) and arsenate were determined to be nonessential activators, characterized by a constant value of (Vmax/Km)app, increasing nitrate reductase activity by weakening nitrate binding without affecting the stability of the transition state. Phosphate had no effect on product inhibition by nitrite (KI = 0.33 mM) or the oxidation-reduction midpoint potentials of the Mo center. PMID- 2550064 TI - Any of several lysines can react with 5'-isothiocyanatofluorescein to inactivate sodium and potassium ion activated adenosinetriphosphatase. AB - Determinations of reaction stoichiometry demonstrate that the covalent incorporation of one molecule of 5'-isothiocyanatofluorescein can inactivate one molecule of sodium and potassium ion activated adenosinetriphosphatase in agreement with earlier determination of this stoichiometry. Several different modified peptides are produced, however, when the modified enzyme is digested with trypsin. One of these peptides has been identified as HLLVMK (thioureidylfluorescein)GAPER by use of a specific immunoadsorbent. The modified lysine is lysine 501 in the amino acid sequence of the alpha polypeptide of (Na+ + K+)-ATPase. This peptide has been previously isolated from such digests [Farley, R. A., Tran, C. M., Carilli, C. T., Hawke, D., & Shively, J. E. (1984) J. Biol. Chem. 259, 9532-9535]. The other specifically modified peptides have been purified and identified by amino acid sequencing. Their sequences identify lysine 480 and lysine 766 from the alpha polypeptide as amino acids modified by 5'-isothiocyanatofluorescein in reactions sensitive to the addition of ATP and responsible for inactivation of the enzyme. PMID- 2550065 TI - Electron-nuclear double resonance spectroscopy of the desulfo-inhibited molybdenum(V) center in bovine milk xanthine oxidase. AB - The "desulfo-inhibited" Mo(V) center of bovine milk xanthine oxidase has been investigated by electron-nuclear double resonance spectroscopy. Comparison of spectral data obtained from samples prepared with [1H4]ethylene glycol and with [2H4]ethylene glycol allowed assignment of proton resonance lines due to the methylene protons of the coordinated ethylene glycol (AH = 3.6 MHz). Deuterium resonance lines were observed with the deuterated sample (AD = 0.4 MHz). No spectral evidence was obtained for any weakly coupled nitrogen nuclei to the Mo center under a variety of conditions. Dissolution of the sample in D2O had little effect on the resonance lines centered about the proton Zeeman frequency, which shows they are not due to exchangeable protons and suggests the Mo center does not have contact with bulk solvent. A deuterium delta m = +/- 2 "forbidden" transition is observed at high radio-frequency power levels, which suggests either an exchangeable proton on a Mo ligand or a coordinated solvent. Weakly coupled, nonexchangeable proton lines are observed about the free proton frequency, which exhibit properties characteristic of alpha-protons. A number of arguments are presented to support the proposal that these protons originate from the C(1') and C(2') positions on the side chain of the molybdopterin cofactor. PMID- 2550066 TI - Analysis of the relative contributions of the nuclear Overhauser interproton distance restraints and the empirical energy function in the calculation of oligonucleotide structures using restrained molecular dynamics. AB - The relative contributions of the interproton distance restraints derived from nuclear Overhauser enhancement measurements and of the empirical energy function in the determination of oligonucleotide structures by restrained molecular dynamics are investigated. The calculations are based on 102 intraresidue and 126 interresidue interproton distance restraints derived from short mixing time two dimensional nuclear Overhauser enhancement data on the dodecamer 5'd(CGCGPATTCGCG)2 [Clore, G.M., Oschkinat, H., McLaughlin, L.W., Benseler, F., Scalfi Happ, C., Happ, E., & Gronenborn, A.M. (1988) Biochemistry 27, 4185-4197]. Eight interproton distance restraint lists were made up with errors ranging from 0.1/+0.2 to -1.2/+1.3 A for r less than 2.5 A and from -0.2/+0.3 to -1.3/+1.4 A for r greater than or equal to 2.5 A. These restraints were incorporated into the total energy function of the system in the form of square-well potentials with force constants set sufficiently high to ensure that the deviations between calculated distances and experimental restraints were very small (average interproton distance rms deviation of less than 0.06 A). For each data set, six calculations were carried out, three starting from classical A-DNA and three from classical B-DNA. The results show that structural changes occurring during the course of restrained molecular dynamics and the degree of structural convergence are determined by the interproton distance restraints. All the structures display similar small deviations from idealized geometry and have the same values for the nonbonding energy terms comprising van der Waals, electrostatic, and hydrogen bonding components. Thus, the function of the empirical energy function is to maintain near perfect stereochemistry and nonbonded interactions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550067 TI - Affinity purification of the voltage-sensitive sodium channel from electroplax with resins selective for sialic acid. AB - The voltage-sensitive sodium channel present in the eel (Electrophorus electricus) has an unusually high content of sialic acid, including alpha-(2--- 8)-linked polysialic acid, not found in other electroplax membrane glycopeptides. Lectins from Limax flavus (LFA) and wheat germ (WGA) proved the most effective of 11 lectin resins tried. The most selective resin was prepared from IgM antibodies against Neisseria meningitidis alpha-(2----8)-polysialic acid which were affinity purified and coupled to Sepharose 4B. The sodium channel was found to bind to WGA, LFA, and IgM resins and was readily eluted with the appropriate soluble carbohydrates. Experiments with LFA and IgM resins demonstrated binding and unbinding rates and displacement kinetics, which suggest highly specific binding at multiple sites on the sodium channel protein. In preparative-scale purification of protein previously fractionated by anion-exchange chromatography, without stabilizing TTX, high yields were reproducibly obtained. Further, when detergent extracts were prepared from electroplax membranes fractionated by low speed sedimentation, a single step over the IgM resin provided a 70-fold purification, yielding specific activities of 3200 pmol of [3H]TTX-binding sites/mg of protein and a single polypeptide of approximately 285,000 Da on SDS acrylamide gels. No small peptides were observed after this 5-h isolation. We further describe a cation-dependent stabilization with millimolar levels of monovalent and micromolar levels of divalent species. PMID- 2550068 TI - Isolation and characterization of one soluble and two membrane-associated forms of phosphoinositide-specific phospholipase C from human platelets. AB - Two forms (mPLC-I, mPLC-II) of phosphoinositide-specific phospholipase C have been purified, 1494- and 1635-fold, respectively, from plasma membranes of human platelets. Purified mPLC-I and mPLC-II had estimated molecular weights by gel filtration and sodium dodecyl sulfate-polyacrylamide gels of 69,000 and 63,000, respectively. Two cytosolic forms (PLC-I and PLC-II) of phosphoinositide-specific phospholipase C were also resolved on a phenyl-Sepharose column. The major cytosolic form present in outdated platelets, PLC-II, was purified to homogeneity by chromatography on Fast Q-Sepharose, cellulose phosphate, heparin-agarose, phenyl-Sepharose, Superose 12, DEAE-5PW, and hydroxylapatite. Purified PLC-II had a molecular weight of 57,000 on sodium dodecyl sulfate-polyacrylamide gels. mPLC I, mPLC-II, and PLC-II hydrolyzed both PI and PIP2. The Vmax for PIP2 hydrolysis was similar for all three forms of PLC and was approximately 5-fold greater than for PI hydrolysis. The Km for PIP2 hydrolysis was also similar for the three enzymes. In contrast, the Km for PI hydrolysis by PLC-II was 10-fold lower than by mPLC-I and mPLC-II. In addition, antibody prepared against PLC-II did not cross-react with either mPLC-I or mPLC-II. These data indicate that platelets contain membrane-associated phosphoinositide-specific phospholipases C that are distinct from at least one cytosolic form (PLC-II) of the enzyme. PMID- 2550069 TI - A novel, arsenite-sensitive E2 of the ubiquitin pathway: purification and properties. AB - In the multienzyme ubiquitin-dependent proteolytic pathway, conjugation of ubiquitin to target proteins serves as a signal for protein degradation. Rabbit reticulocytes possess a family of proteins, known as E2's, that form labile ubiquitin adducts by undergoing transthiolation with the ubiquitin thiol ester form of ubiquitin activating enzyme (E1). Only one E2 appears to function in ubiquitin-dependent protein degradation. The others have been postulated to function in regulatory ubiquitin conjugation. We have purified and characterized a previously undescribed E2 from rabbit reticulocytes. E2(230K) is an apparent monomer with a molecular mass of 230 kDa. The enzyme forms a labile ubiquitin adduct in the presence of E1, ubiquitin, and MgATP and catalyzes conjugation of ubiquitin to protein substrates. Exogenous protein substrates included yeast cytochrome c(Km = 125 mu M; kcat approximately 0.37 min-1) and histone H3 (Km less than 1.3 mu M; kcat approximately 0.18 min-1) as well as lysozyme, alpha lactalbumin, and alpha-casein. E2(230K) did not efficiently reconstitute Ub dependent degradation of substrates that it conjugated, either in the absence or in the presence of the ubiquitin-protein ligase that is involved in degradation. E2(230K) may thus be an enzyme that functions in regulatory Ub conjugation. Relative to other E2's, which are very iodoacetamide sensitive, E2(230K) was more slowly inactivated by iodoacetamide (k(obs) = 0.037 min-1 at 1.5 mM iodoacetamide; pH 7.0, 37 degrees C). E2(230K) was also unique among E2's in being subject to inactivation by inorganic arsenite (k(i)max = 0.12 min-1; K(0.5) = 3.3 mM; pH 7.0, 37 degrees C). Arsenite is considered to be a reagent specific for vicinal sulfhydryl sites in proteins, and inhibition is usually rapidly reversed upon addition of competitive dithiol compounds. Inactivation of E2(230K) by arsenite was not reversed within 10 min after addition of dithiothreitol at a concentration that blocked inactivation if it was premixed with arsenite; inactivation is therefore irreversible or very slowly reversible. We postulate that a conformation change of E2(230K) may be rate-limiting for interaction of enzyme thiol groups with arsenite. PMID- 2550070 TI - Predicted structures of the cGMP binding domains of the cGMP-dependent protein kinase: a key alanine/threonine difference in evolutionary divergence of cAMP and cGMP binding sites. AB - Mammalian cGMP- and cAMP-dependent protein kinase show considerable similarity in amino acid sequence, although they specifically bind different cyclic nucleotides. Results of cGMP analogue binding experiments, combined with modeling of the cGMP binding sites by analogy to the structure of the homologous catabolite gene activator protein, suggest that a threonine residue forms a hydrogen bond with the 2-NH2 of cGMP. This threonine is invariant in all cGMP binding domains, but the corresponding residue in 23 out of 24 cAMP binding sites of protein kinases is alanine, which cannot form the same hydrogen bond. This alanine/threonine difference has the potential for discriminating between cAMP and cGMP and may be important in the evolutionary divergence of cyclic nucleotide binding sites. PMID- 2550071 TI - Commentary on 'New Methods for the Study of the Carbon Monoxide Compounds of Respiratory Enzymes'. PMID- 2550072 TI - New methods for the study of the carbon monoxide compounds of respiratory enzymes. 1953. PMID- 2550073 TI - The identification of the sodium-pump as the membrane-bound Na+/K+-ATPase: a commentary on 'The Influence of Some Cations on an Adenosine Triphosphatase from Peripheral Nerves'. PMID- 2550074 TI - The influence of some cations on an adenosine triphosphatase from peripheral nerves. 1957. PMID- 2550075 TI - Effect of 3,4-dihydroxybutyl-1-phosphonate on cardiolipin synthesis in B. subtilis. AB - Endogenous phosphatidylglycerol is rapidly transformed into cardiolipin when B. subtilis 168 cells were incubated in a buffer without an energy source. Upon addition of 3,4-dihydroxybutyl-1-phosphonate (DHBP), a synthetic glycerol 3 phosphate analogue, this synthesis was completely blocked after a short lag; if the cells were grown in the presence of the analogue, there was no lag. When membrane fractions were incubated with exogenous [32P]phosphatidylglycerol, free DHBP and glycerol 3-phosphate had no effect on [32P]cardiolipin synthesis, but phosphatidyl-DHBP and phosphatidylglycerolphosphate were potent inhibitors. These results are consistent with our hypothesis that phosphatidylglycerolphosphate, the phosphatidylglycerol precursor, might also be a physical inhibitor of cardiolipin synthesis. PMID- 2550076 TI - Metabolism of pyrenedecanoic acid in Epstein-Barr virus-transformed lymphoid cell lines from normal subjects and from a patient with multisystemic lipid storage myopathy. AB - A lymphoid cell line has been established from a patient with multisystemic lipid storage myopathy and showed a major triacylglycerol storage, whereas the content of other neutral lipids and phospholipids was in the normal range. The metabolism of the triacylglycerols has been investigated in this lymphoid cell line from multisystemic lipid storage myopathy as well as in control cells through pulse chase experiments using 10-(1-pyrene)decanoic acid (P10), a fluorescent fatty acid derivative, as precursor. After 1 h incubation, the uptake of P10 was not significantly different in multisystemic lipid storage myopathy and control lymphoid cells. The amount of fluorescent lipids synthesized by the lymphoid cells was proportional to the concentration of P10 in the culture medium. After 24 h incubation, at any extracellular concentration of P10, the content of P10 labelled triacylglycerols was much higher in multisystemic lipid storage myopathy cells than in controls. Chase experiments showed an impairment in the rate of degradation of biosynthesized triacylglycerols in multisystemic lipid storage myopathy lymphoblasts compared to controls with time of chase (the ratio P10 triacylglycerols/P10-phospholipids increased in mutant cells while it decreased in normal cells). Elsewhere, no enzyme deficiency of the neutral triacylglycerol lipase activity, has been found in multisystemic lipid storage myopathy lymphoid cells. PMID- 2550077 TI - Polarized infrared absorption of Na+/K+-ATPase studied by attenuated total reflection spectroscopy. AB - Na+/K+-ATPase can be isolated from the outer medulla of mammalian kidney in the form of flat membrane fragments containing the enzyme in a density of 10(3)-10(4) protein molecules per microm2 (Deguchi et al. (1977) J. Cell. Biol. 75, 619-634). In this paper we show that these membrane fragments can be bound to a germanium plate coated with a phospholipid bilayer. With this system infrared spectroscopic studies of the enzyme have been carried out using the technique of attenuated total reflection (ATR). At a coverage of the lipid surface corresponding to 30 40% of a monolayer of membrane fragments, characteristic infrared bands of the protein such as the amide I and II bands can be resolved. About 24% of the NH groups of the peptide backbone are found to be resistant to proton/deuterium exchange within a time period of several days. Evidence for orientation of the protein with respect to the supporting lipid layer is obtained from experiments with polarized light, the largest polarization effects being associated with the COO- band at 1400 cm-1. Experiments with aqueous media of different ionic composition indicate that the average orientation of transition moments changes when K+ in the medium is replaced by Tris+ or Na+. PMID- 2550078 TI - Effect of calcium on the interactions between Ca2+-ATPase molecules in sarcoplasmic reticulum. AB - The interaction between Ca2+-ATPase molecules in the native sarcoplasmic reticulum membrane and in detergent solutions was analyzed by chemical crosslinking, high performance liquid chromatography (HPLC), and by the polarization of fluorescence of fluorescein 5'-isothiocyanate (FITC) covalently attached to the Ca2+-ATPase. Reaction of sarcoplasmic reticulum vesicles with glutaraldehyde causes the crosslinking of Ca2+-ATPase molecules with the formation of dimers, tetramers and higher oligomers. At moderate concentrations of glutaraldehyde solubilization of sarcoplasmic reticulum by C12 E8 or Brij 36T (approximately equal to 4 mg/mg protein) decreased the formation of higher oligomers without significant interference with the appearance of crosslinked ATPase dimers. These observations are consistent with the existence of Ca2+ ATPase dimers in detergent-solubilized sarcoplasmic reticulum. Ca2+ (2-20 mM) and glycerol (10-20%) increased the degree of crosslinking at pH 6.0 both in vesicular and in solubilized sarcoplasmic reticulum, presumably by promoting interactions between ATPase molecules; at pH 7.5 the effect of Ca2+ was less pronounced. In agreement with these observations, high performance liquid chromatography of sarcoplasmic reticulum proteins solubilized by Brij 36T or C12 E10 revealed the presence of components with the expected elution characteristics of Ca2+-ATPase oligomers. The polarization of fluorescence of FITC covalently attached to the Ca2+-ATPase is low in the native sarcoplasmic reticulum due to energy transfer, consistent with the existence of ATPase oligomers (Highsmith, S. and Cohen, J.A. (1987) Biochemistry 26, 154-161); upon solubilization of the sarcoplasmic reticulum by detergents, the polarization of fluorescence increased due to dissociation of ATPase oligomers. Based on its effects on the fluorescence of FITC-ATPase, Ca2+ promoted the interaction between ATPase molecules, both in the native membrane and in detergent solutions. PMID- 2550079 TI - Incompletely processed LH molecules synthesized by rat gonadotrophs treated with inhibitors of oligosaccharide processing. AB - Inhibitors of N-linked oligosaccharide processing are useful tools for studies on the biological function of the oligosaccharide structures in glycoprotein hormones. We have synthesized molecules of lutropin (LH) containing high-mannose- and hybrid-type oligosaccharides using rat gonadotroph-enriched primary cultures in the presence of castanospermine (a glucosidase I inhibitor) or swainsonine (a mannosidase II inhibitor), in order to compare the actions of these molecules with that of the hormone containing complex-type oligosaccharides in the activation of the receptor-adenylate cyclase system. Treatment of gonadotrophs with the above inhibitors caused an increase in the synthesis of highly basic LH molecules (pI 9.6-10.0), because addition of charged carbohydrate moieties to these molecules was prevented. Characterization of the oligosaccharide structure performed by enzymatic treatment (endoglycosidase H and neuraminidase) and the use of immobilized lectins (wheat germ agglutinin and Ricinus communis agglutinin 120) showed that these inhibitor-synthesized LH molecules contained high-mannose- and hybrid-type (asialo and sialylated) oligosaccharides. Their immunological properties were similar to that of complex-type oligosaccharide LH, but they had significantly higher receptor-binding ability in comparison with a sialylated complex-type oligosaccharide LH (about 12-fold) and an asialo complex-type oligosaccharide LH (about 3-fold). It was noted that the incompletely processed molecules were less potent than complex-type oligosaccharide LH in the activation of adenylate cyclase of Leydig cells, showing about 40-60% of the activity induced by the sialylated complex-type oligosaccharide molecule. The present data indicate that the inhibition of terminal processing of N-linked oligosaccharides by castanospermine and swainsonine impairs the full hormonal function of rat LH. PMID- 2550080 TI - Purification of intrinsic factor receptor from pig ileum using as affinity medium human intrinsic factor covalently bound to Sepharose. AB - Intrinsic factor receptor was purified from hog ileum using human intrinsic factor covalently bound to Sepharose. A yield of 49.6% and a specific activity of about 2500 pmol/mg protein were achieved. The purified receptor was very unstable: 24 h of storage or addition of sodium phosphate precipitated it. The association constant of the receptor for the cyano[57Co]cobalamin-intrinsic factor complex was estimated to be 2.1 nM-1. In native polyacrylamide gel electrophoresis it resolved in two 256 and 320 kDa bands; beta-mercaptoethanol treatment cleared it into four bands corresponding to molecular masses of 107, 81.8, 63.5 and 53.2 kDa. An additional 39.3 kDa band was considered to be an artefact due to the presence of Triton X-114. Isoelectric focusing polyacrylamide gel electrophoresis resolved the receptor into two isoproteins isoelectric at pH 4.7 and 5.1. A similar result was obtained in column electrofocusing with the 125I-iodinated receptor. The 125I-labelled receptor did not crossreact with rabbit anti-human intrinsic factor antiserum. The electrophoretic properties of the receptor purified with intrinsic factor covalently bound to Sepharose were compared to those of the receptor purified by the use of the classical cobalamin affinity medium. It was concluded that a disassembled receptor was produced using the classical method. PMID- 2550081 TI - Free-radical formation by mitomycin C and its novel analogs in cardiac microsomes and the perfused rat heart. AB - Using a spin-trapping technique, we have examined free-radical formation by mitomycin C and its analogs, BMY 25282 and BMY 25067, in rat cardiac microsomes and isolated perfused rat hearts. All three drugs stimulated 2--4-fold OH radical formation in cardiac microsomes which was inhibited by SOD and catalase. Superoxide anion radical was also detected in the presence of diethylenetetraaminopentaacetic acid. Addition of DMSO yielded methyl radicals, thus indicating the production of free OH under these conditions. Similar stimulation of OH formation (2--3-fold) in the perfusates from rat hearts was detected with all three drugs. Perfusion with catalase (550 U/ml) completely suppressed the OH signal both in the presence and absence of the drugs, thus suggesting the intermediacy of hydrogen peroxide. However, BMY 25067-induced OH formation was more sensitive to inhibition by superoxide dismutase (SOD) and the iron chelator ICRF-187. Perfusion with DMSO produced methyl radicals at the expense of OH in the presence of all three drugs. SOD and catalase inhibited DMPO OH signals, indicating that most of the OH formation was extracellular in this setting. While mitomycin C and BMY 25067 (up to 10 microM) did not affect the heart rate, perfusion with 10 microM BMY 25282 caused acute arrhythmia and cardiac standstill within 20 min. An initial surge in OH formation (2-fold) accompanied this cardiotoxic effect. Both the arrhythmia and the free radical signal were partially blocked by SOD, catalase and ICRF-187, indicating that iron dependent oxygen radical formation from BMY-25282 (and possibly other compounds) is involved, in part, in inducing toxic manifestations in the rat heart and possibly in clinic. PMID- 2550082 TI - The enzymatic evaluation of procollagenase and collagenase inhibitors in crude biological media. AB - The validity of the enzymatic assay of procollagenase within crude biological media containing also the collagenase inhibitor TIMP (tissue inhibitor of metalloproteinases) as well as other (pro)metalloproteinases and sometimes, metalloproteinase-TIMP complexes, has been reevaluated. To be enzymatically assayed, procollagenase has to be activated. The standard activation procedures by either trypsin or 4-aminophenylmercuric acetate (APMA) both allow an optimal recovery of collagenase from procollagenase when the media do not contain free TIMP. However, they do not destroy TIMP nor do they reactivate the collagenase present in enzyme-inhibitor complexes. Therefore, the collagenase formed by the activation of procollagenase in the presence of free TIMP is immediately inactivated by binding to the inhibitor. As a result, both the bound collagenase and TIMP can no longer be assayed by enzymatic methods. An optimal recovery of collagenase can, however, be obtained if free TIMP is neutralized by the binding of other tissue metalloproteinases (such as those present in culture media of rabbit bone marrow-derived macrophages) prior to the activation and assay of procollagenase. Similarly, it is possible to recover under an active free form a large part of the TIMP present in collagenase- (or other metalloproteinase-)TIMP complexes by heating the complexes at acid pH under conditions which inactivate the collagenase. PMID- 2550083 TI - Evidence for the direct interaction of chicken gizzard 5'-nucleotidase with laminin and fibronectin. AB - The ectoenzyme 5'-nucleotidase purified from chicken gizzard is shown to specifically interact with laminin and fibronectin, components of the extracellular matrix, by a number of different techniques: (i) cosedimentation with laminin by sucrose gradient centrifugation; (ii) affinity adsorption to both laminin- and fibronectin-Sepharose 4-B; (iii) specific binding to both laminin and fibronectin dotted onto cellulose filters; and (iv) monoclonal antibodies against 5'-nucleotidase are shown to interfere with the interaction of 5' nucleotidase with laminin and fibronectin. For all the techniques employed, the interactions were found to be specific, since 5'-nucleotidase did not bind to unrelated proteins such as bovine serum albumin or to monomeric actin. The interaction of purified chicken gizzard 5'-nucleotidase could be demonstrated for the hydrophobic enzyme solubilized in detergent and after its reconstitution into artificial phospholipid vesicles. The affinity adsorption experiments indicate that reconstituted enzyme binds more strongly to both laminin and fibronectin. The 5'-nucleotidase employed in this study is anchored to the plasma membrane by a glycan-phosphatidylinositol linker. After treatment with phosphatidylinositol specific phospholipase C, the enzyme is transformed into a hydrophilic form, for which interactions with laminin and fibronectin could also be demonstrated by the dot-blot technique. Thus controlled cleavage of the phosphatidylinositol linker of 5'-nucleotidase could enable cells to rapidly alter their adhesiveness to certain components of the extracellular matrix. PMID- 2550084 TI - Purification of basic fibroblast growth factor receptors from bovine brain. AB - Fibroblast growth factors are proteins which play a major role, in vitro and in vivo, in the control of cellular growth and differentiation of a large number of cells. Biological activities of these factors are mediated by the interaction with specific membrane receptors. Previous studies indicated that the apparent molecular weight of a family of these receptors for the basic form of Fibroblast Growth Factor (bFGF), ranges from 125 to 165 kDa according to cell species and types. We have purified this family of receptors from bovine brain. We first set up a radioreceptor assay to detect receptors throughout the purification by measuring its ability to inhibit the fixation of radiolabeled bFGF to insolubilized membranes from bovine brain. The purification was also monitored by using cross-linking reagents in order to allow the visualization of radiolabeled bFGF bound to its receptor. The first purification steps involved 2 anion exchange chromatographic steps, DEAE Trysacryl and FPLC Mono Q, and yielded an enrichment over 500 fold. Affinity chromatography with bFGF immobilized on Sepharose 4B was then performed. Covalent fixation of bFGF to the Sepharose matrix was carried out in presence of N-acetylated heparin in order to protect the recognition site for bFGF on its receptor. These 3 chromatographic steps yielded only 2 bands of apparent molecular weight of 100 kDa and 135 kDa as detected by electrophoresis. These 2 bands are also detected after chromatography on immobilized wheat germ agglutinin hence confirming the presence of carbohydrates on bFGF receptors. PMID- 2550085 TI - Neural integration by short term potentiation. AB - Neurophysiological studies in the oculomotor system suggest that an integrative operation is required in order to derive an eye position signal from a command signal which usually correlates with eye velocity. Several proposed models for a neural integrator are examined. All these models incorporate some form of positive feedback as a basic mechanism. Based on the performance of the models, we argue that such a scheme require extreme high precision in order to work properly. A new model based on potentiation phenomena in synaptic transmission is proposed and is shown to be free from the deficits of most previous models. The proposed model also accounts for various neural behaviors in a very natural way. A possible implementation of the model is also discussed in the context of the vestibulo-ocular reflex (VOR). PMID- 2550086 TI - Influence of follicular maturation on luteinizing hormone and guanosine 5'-O thiotriphosphate-promoted breakdown of phosphoinositides and calcium mobilization in chicken granulosa cells. AB - Previous studies in this laboratory have shown a remarkable increase in the capacity of avian granulosa cells to produce progesterone during the last 2-3 days of folliculogenesis, with concomitant increases in the activities of key steroidogenic enzymes. In view of the proposed involvement of inositol phospholipids and their hydrolytic products in signal transduction in steroidogenic cells, we investigated in this study the influence of follicular maturation on phosphoinositide-specific phospholipase C (PLC) and intracellular Ca2+ release in chicken granulosa cells. Resting concentrations of intracellular calcium ([Ca2+]i) as measured in Fura 2/AM-loaded cells increased significantly during the last 48 h of follicular maturation, from 185 +/- 9 nM in the third largest follicle (F3) to 355 +/- 26 in the cells of the largest (F1) follicle. Luteinizing hormone (LH) caused a dose-related rise in [Ca2+]i, but the dose response was left-shifted by more than one order of magnitude in F1 cells compared to F2 cells. In granulosa cells of less developed follicles, LH failed to raise [Ca2+]i. To assess PLC activity, granulosa cells from F1, F2, and F3 follicles were labeled with [3H]inositol for 2 h and then stimulated with LH (0.1 microgram/ml). Time course studies showed that within 30 s, phosphatidylinositol 4,5 bisphosphate (PIP2) decreased by 33%, 13%, and 11% in F1, F2, and F3 cells, respectively. Similar responses were obtained when permeabilized cells were exposed to guanosine 5'-O-thiotriphosphate, which also caused a corresponding increase in 45Ca efflux from F1 and F2 cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550087 TI - Functional relationship between receptor binding and biological activity for analogs of mammalian and salmon gonadotropin-releasing hormones in the pituitary of goldfish (Carassius auratus). AB - The relationship between gonadotropin-releasing hormone (GnRH) receptor binding and biological activity in the goldfish pituitary for mammalian and salmon GnRH (sGnRH) analogs with structural modification at the C terminus involving replacement of glycine amide with an alkyl amine and replacement of the Gly6 residue with D amino acids was examined. The GnRH receptor binding data were analyzed with a computerized curve-fitting program (LIGAND) for a single as well as two classes of binding sites; analysis based on one site fit estimated binding affinity and capacity for one class of binding site, and analysis based on two site fit estimated binding affinity and capacity for two classes of binding sites (high-affinity/low-capacity and low-affinity/high-capacity binding sites). The estimated receptor affinity values were then used to determine the correlation between binding affinity and gonadotropin (GTH)-release potency in vitro. The highest correlation between biological activity and receptor binding affinity was obtained for the high-affinity/low-capacity binding sites and GnRH analogs containing Trp7 and Leu8 residues (i.e., the salmon GnRH structural format) (R = 0.940 +/- 0.150). For the same group of GnRH analogs, there was no significant correlation between the relative GTH-release potency and binding affinity of the low-affinity/high-capacity sites (R = 0.159 +/- 0.434), or that obtained from a one-site fit (R = 0.198 +/- 0.431). Similarly, for mammalian GnRH analogs, significant correlation between binding affinity and biological activity (R = 0.406 +/- 0.049) was only obtained for the high-affinity sites, although the degree of correlation was significantly lower than that obtained for salmon GnRH analogs. The present findings provide strong support for the hypothesis that high affinity GnRH receptors are involved in the control of GTH release in the goldfish pituitary. In addition, the results demonstrate clearly that the presence of Trp7, Leu8 residues in salmon GnRH molecule, a native peptide in goldfish, is important for recognition of the ligand by the GnRH receptors in the goldfish pituitary, and that structural modifications at positions 6 and 10 in this peptide can increase receptor binding affinity and biological activity at the pituitary level. The most active sGnRH analog identified to date is [D-Arg6, Pro9-NEt]-sGnRH. PMID- 2550088 TI - Sulfated oviductal glycoproteins in the rabbit: quantitation by competitive enzyme-linked immunosorbent assay. AB - The rabbit oviductal epithelium synthesizes and secretes a family of antigenically related, sulfated oviductal glycoproteins (SOG). Anti-SOG monoclonal antibodies (Mabs) were produced and two (Mab 1 and Mab 2) were selected for further characterization. Periodate oxidation of Western blots of oviductal fluid did not affect the binding of Mab 1 or Mab 2, thus suggesting that these antibodies recognized protein rather than carbohydrate epitopes on SOG. The specificity of Mab 1 was determined by Western blot analysis of tissues obtained from estrous rabbits and from the male rabbit reproductive tract. SOG was identified in tissue extracts of both the oviductal ampulla and isthmus. Cervix was the only non-oviductal tissue with which Mab 1 cross-reacted. Mab 1 was used to isolated SOG from whole oviductal fluid by immuno-affinity chromatography. Affinity-purified SOG and Mab 1 were used to develop a quantitative, SOG-specific, competitive enzyme-linked immunosorbent assay. This assay was used to quantify SOG in rabbit oviductal fluid collected during estrus and pseudopregnancy. SOG secretion during pseudopregnancy was resolved into two transient episodes of increased secretion. Maximum SOG secretion (X = 1039 +/- 199 micrograms/day) occurred within 48 h of the induction of pseudopregnancy. A second period of enhanced SOG secretion (X = 308 +/- 46 micrograms/day) occurred during the fifth and sixth days of pseudopregnancy. Baseline SOG secretion occurred during estrus at approximately 60% of maximum postovulatory secretion. PMID- 2550089 TI - Location of the heme-Fe atoms within the profile structure of a monolayer of cytochrome c bound to the surface of an ultrathin lipid multilayer film. AB - We have recently developed x-ray diffraction methods to derive the profile structure of ultrathin lipid multilayer films having one to five bilayers (e.g., Skita, V., W. Richardson, M. Filipkowski, A.F. Garito, and J.K. Blasie. 1987. J. Physique. 47:1849-1855). Furthermore, we have employed these techniques to determine the location of a monolayer of cytochrome c bound to the carboxyl group surface of various ultrathin lipid multilayer substrates via nonresonance x-ray diffraction (Pachence, J.M., and J.K. Blasie. 1987. Biophys. J. 52:735-747). Here an intense tunable source of x-rays (beam line X9-A at the National Synchrotron Light Source at the Brookhaven National Laboratory) was utilized to measure the resonance x-ray diffraction effect from the heme-Fe atoms within the cytochrome c molecular monolayer located on the carboxyl surface of a five monolayer arachidic acid film. Lamellar x-ray diffraction was recorded for energies above, below, and at the Fe K-absorption edge (E = 7,112 eV). An analysis of the resonance x-ray diffraction effect is presented, whereby the location of the heme-Fe atoms within the electron density profile of the cytochrome c/arachidic acid ultrathin multilayer film is indicated to +/- 3 A accuracy. PMID- 2550090 TI - Electrostatic and steric control of electron self-exchange in cytochromes c, c551, and b5. AB - The ionic strength dependence of the electron self-exchange rate constants of cytochromes c, c551, and b5 has been analyzed in terms of a monopole-dipole formalism (van Leeuwen, J.W. 1983. Biochim. Biophys. Acta. 743:408-421). The dipole moments of the reduced and oxidized forms of Ps. aeruginosa cytochrome c551 are 190 and 210 D, respectively (calculated from the crystal structure). The projections of these on the vector from the center of mass through the exposed heme edge are 120 and 150 D. For cytochrome b5, the dipole moments calculated from the crystal structure are 500 and 460 D for the reduced and oxidized protein; the projections of these dipole moments through the exposed heme edge are -330 and -280 D. A fit of the ionic strength dependence of the electron self exchange rate constants gives -280 (reduced) and -250 (oxidized) D for the center of mass to heme edge vector. The self-exchange rate constants extrapolated to infinite ionic strength of cytochrome c, c551, and b5 are 5.1 x 10(5), 2 x 10(7), and 3.7 x 10(5) M-1 s-1, respectively. The extension of the monopole-dipole approach to other cytochrome-cytochrome electron transfer reactions is discussed. The control of electron transfer by the size and shape of the protein is investigated using a model which accounts for the distance of the heme from each of the surface atoms of the protein. These calculations indicate that the difference between the electrostatically corrected self-exchange rate constants of cytochromes c and c551 is due only in part to the different sizes and heme exposures of the two proteins. PMID- 2550093 TI - Orientational and rotational velocity correlation functions for water-hydrating B and Z-DNA double helices. AB - The molecular dynamics simulations reported earlier for the structure and dynamics of water molecules hydrating B- and Z-DNA double helices are analyzed for the orientational correlation functions and the proton rotational velocity autocorrelation functions. The spectra of the rotational velocity autocorrelation functions obtained from the simulation results are compared with the neutron inelastic scattering experiments on hydrated Na-DNA samples. The results predict a small frequency component associated with water molecules bound to the double helices that disappears for waters away from the double helix. PMID- 2550091 TI - Evidence that the two free sulfhydryl groups of plasma fibronectin are in different local environments. Saturation-recovery electron spin resonance study. AB - Human plasma fibronectin is a dimer consisting of two subunits; each contains two cryptic thiol groups that were selectively labeled with an 15N,2H-maleimide spin label. Previous studies using conventional X-band electron spin resonance (ESR) methods showed that the spectrum of the labeled protein displays a single strongly immobilized component with an effective rotational correlation time of approximately 17 ns, suggesting that the physical environments of the two labeled sites per chain are indistinguishable. Here we have used saturation-recovery ESR to measure directly electron spin-lattice relaxation time (T1) of the labeled protein in solution at 27 degrees C. Interestingly, the time evolution of the signal was found to be biphasic, which was deconvoluted into two T1 values of 1.37 and 4.53 microseconds. Thus, the two spin-labeled sulfhydryl sites of plasma fibronectin (Fn), being similar in rates of rotational diffusion, differ by a factor of 3.2 in T1. Parallel experiments using various fibronectin fragments showed that the 1.37-microseconds component is associated with the label attached onto the thiol located in between the DNA-binding and the cell-binding domains, and the 4.53-microseconds component is associated with the label attached onto the thiol located within the carboxyl-terminal fibrin-binding domain. The data suggest that the saturation-recovery ESR is a useful method for differentiating multiple spin-labeled sites on macromolecules in which the labels undergo similar rates of rotational motion. PMID- 2550092 TI - QX-314 restores gating charge immobilization abolished by chloramine-T treatment in squid giant axons. AB - The gating status of the QX-314 bound Na channels before and after suppressing the fast inactivation by chloramine-T (CT) was investigated by studying the gating charge immobilization using the OFF gating current (Ig,OFF). CT treatment, which abolishes the charge immobilization induced by a prolonged depolarization, altered the kinetics of Ig,OFF: the fast phase became insensitive to the pulse duration and the slow phase became three times faster than the control one. However, internally applied QX-314 (in the presence of external TTX) caused an immediate charge immobilization similar to that observed in the absence of CT treatment. The Ig,OFF exhibited kinetics similar to the inactivated channels, decaying with a very fast time course. We conclude that the charge immobilization is restored by QX-314 in the chloramine-T-treated axon and that the gating state of the QX-314-bound channel is similar to the inactivated one. The role of the gating charge immobilization in the use-dependent block mechanism is discussed. PMID- 2550094 TI - Are sexually transmitted papilloma viruses exclusive, necessary and sufficient causative factors of genital organ invasive or in situ neoplasia? PMID- 2550095 TI - Carcinoma of the cervix and smoking. AB - There is considerable evidence from epidemiologic and clinical studies that cigarette smoking is associated with the risk of cervical cancer. Definitive clarification of whether this association is causal will likely have to await definitive identification of the sexually transmitted agent which is probably the most important cause of cervical cancer. Only then will it be possible to clarify the contributions of risk factors with weaker associations with cervical cancer, such as cigarette smoking and socioeconomic status. PMID- 2550096 TI - Sexually transmitted cancers and viruses. AB - Both cervical cancer and its precancerous state cervical intraepithelial neoplasia (CIN) have the characteristics of being sexually transmitted. Formerly herpes simplex virus (HSV) but more recently human papillomavirus (HPV) which both infect the cervix have been implicated in causation. The role of these viruses as possible initiators of cancer or as potential cofactors of cocarcinogens is discussed at the molecular level within the context of the disease process. PMID- 2550097 TI - Cervical carcinoma and human cytomegalovirus. AB - Human cytomegalovirus (HCMV) is a sexually transmitted virus which has been implicated in both cervical cancer and its precancerous state, cervical intraepithelial neoplasia (CIN). The evidence for this and the current possible mechanisms by which HCMV could be involved are discussed at the molecular level. PMID- 2550098 TI - Regression of the malignant aspects of intraepithelial neoplasias following an LH RH agonist treatment and detection of human papillomavirus by molecular hybridization. AB - Patients presenting genital intraepithelial neoplasia and/or flat condyloma were treated with DTrp6-LH-RH (triptorelin) to induce a transitory suppression of estrogens. This treatment led in some cases to a complete clinical and histological regression accompanied by a disappearance of human papillomavirus sequences as detected by molecular hybridization. PMID- 2550099 TI - Collagen binding proteins derived from the embryonic fibroblast cell surface recognize arginine-glycine-aspartic acid. AB - Several cell surface proteins (Mr = 120,000, 90,000, 63,000 and 47,000) apparently integral to embryonic fibroblast plasma membranes were extracted with detergent and isolated by collagen affinity chromatography. Certain of these proteins (Mr = 120,000, 90,000, and 47,000) were specifically eluted from collagen affinity columns by synthetic peptides containing the amino acid sequence arginyl-glycyl-aspartic acid (RGD). These data show that a number of collagen binding proteins exist on the embryonic fibroblast cell surface. Some of the proteins may be collagen receptors binding to RGD sequences in the collagen molecule while at least one of the proteins (Mr = 63,000) recognizes features other than RGD. PMID- 2550100 TI - Erythrocyte membrane (Ca2+ + Mg2+)-ATPase in human protein-energy malnutrition. AB - Calmodulin-free ghost membranes were prepared from erythrocytes of kwashiorkor children and from healthy children in the same age bracket. In the absence of calmodulin, the specific activity of Mg2+-dependent Ca2+-pumping ATPase (Ca2+ + Mg2+-ATPase) of kwashiorkor membranes was more than 40 percent lower than the specific activity of the normal enzymes, whose maximum velocity was increased by at least four-fold by the modulator protein. In contrast, the maximum velocity of the enzymes of kwashiorkor membranes was enhanced by calmodulin by about 1 1/2 times the basal activity of the normal enzymes and by 2 times the basal activity of the kwashiorkor enzymes. The affinity of the pump for ATP was lower in the membranes of kwashiorkor children (Km for ATP = 30.6 +/- 2.8 microM ATP) in comparison to normal membranes (Km for ATP = 21.7 +/- 2.0 microM ATP). Similarly, calmodulin-affinity of the enzymes, was lower in kwashiorkor membranes than in the normal membranes irrespective of source of calmodulin. Calmodulin from haemolysates of kwashiorkor red cells activated the enzymes of normal and kwashiorkor membranes to the same degree as calmodulin partially purified from the haemolysate of healthy children. A determination of the dependence of the activity of the pump on calcium in the absence and presence of calmodulin reveals that the affinity of the kwashiorkor enzymes for Ca2+ is at least 70 percent lower than that of enzymes of normal membranes. Altogether, these findings suggest that the Ca2+-pumping ATPase of kwashiorkor membranes is less functional than the enzymes of healthy erythrocytes. PMID- 2550101 TI - Bernard-Soulier syndrome. AB - Bernard-Soulier syndrome (BSS) is a rare autosomal bleeding disorder characterized clinically by prolonged skin bleeding time, normal clot retraction and thrombocytopenia with large and morphologically abnormal platelets, and biochemically by the absence of platelet membrane glycoproteins (GP) Ib, V and IX. GP Ib and GP IX exist in the platelet membrane as a heterodimer complex which acts as the major receptor mediating platelet adhesion to blood vessel subendothelium. Studies with BSS platelets have proved particularly rewarding in the investigation of the GP Ib-IX complex as a multifunctional receptor protein. The transmembrane complex contains binding domains for von Willebrand factor, thrombin, fibrin and quinine/quinidine drug-dependent antibodies as well as an attachment site on the cytoplasmic side of the membrane for a platelet cytoskeleton. In addition, the internal segment of the beta-chain of GP Ib contains a cyclic AMP-dependent protein kinase-associated phosphorylation site which appears to regulate platelet reactivity. Limited proteolytic cleavage of the complex, in particular the GP Ib alpha-chain, has allowed immunological and functional characterization of three distinct domains; a 45 kDa segment at the N terminal end of the alpha-chain of GP Ib, which contains binding sites for von Willebrand factor and thrombin, a 90 kDa highly glycosylated region of GP Ib alpha and a membrane-associated region consisting of the remnant of GP Ib alpha disulphide-linked to GP Ib beta and non-covalently-complexed with GP IX. This membrane-associated region contains the antigenic epitope(s) for quinine/quinidine drug-dependent antibodies. It is highly probable that the future study of platelets from patients with the Bernard-Soulier syndrome will further clarify the role of the GP Ib-IX complex in platelet physiology. PMID- 2550102 TI - Limited small cell lung cancer: possible prognostic impact of initial chemotherapy doses. AB - While the effect of chemotherapy dose on tumor response in small cell lung cancer has been fairly well established, the effect on survival has been retrospectively analyzed only in some series. This particular point was studied in a series of 52 consecutive patients with limited small cell lung cancer treated by an alternating radiotherapy-chemotherapy schedule. The induction treatment consisted of 6 chemotherapy cycles (the planned doses were: doxorubicin 40 mg/m2 day 1, VP16213 75 mg/m2 days 1-3, cyclophosphamide 300 mg/m2 days 3-6, and cisplatinum 100 mg/m2 day 2) alternated after the first 2 cycles with 3 courses of thoracic radiotherapy delivering a total dose of 55 Gy. Eighty-one percent of patients went into complete remission and the 3-year relapse-free survival was 24%. A multivariate analysis of prognostic factors took into account age, sex, T stage, performance status, delayed hematological toxicity to the first course of chemotherapy, actual dose/m2 of each drug during the first course and mean dose/course delivered during the induction treatment after the first cycle of chemotherapy. It was possible to identify 3 independent factors influencing overall survival and relapse-free survival: actual initial dose of cisplatinum, actual initial dose of cyclophosphamide and the T stage. The effect of the initial dose of cisplatinum and cyclophosphamide proved to be linear on relapse free survival. The results of this analysis show a possible effect of initial doses of chemotherapy in the management of limited small cell lung cancer in terms of both distant metastasis and overall survival rates. PMID- 2550103 TI - [Clinically impalpable mammary carcinoma: morphologic and histogenetic analysis. Apropos of 114 cases]. AB - In a population of 2,372 consecutive cases of breast carcinomas, 114 cases of clinically occult non palpable breast lesions have been diagnosed (4.8%). 51% of them can be considered as minimal breast carcinomas (MBC) by Gallager's definition and 72% by that of the American College of Surgeons; whatever the definition this category has an excellent prognosis with 11% of axillary invasion for the infiltrating tumors under 5 mm and 7% for these under 10 mm and 100% 5 year survival rate in both cases. The category of infiltrating tumors of over 5 mm and 10 mm also has a good prognosis with 21% and 26% of axillary lymph node invasion respectively, with a survival rate of 83.82% and 77.92%. The comparative histological analysis shows at this stage an increase in the intraductal carcinomas (IC) (35% instead of 6% for the palpable carcinomas), the infiltrating ductal carcinomas with predominant intraductal component (IDCPIC) (19% instead of 12%) and the tubular carcinomas (11% instead of 3%). The study of the peritumoral and environmental mastopathy and the histological repartition, confirms the classical histogenetic arguments regarding breast carcinomas. PMID- 2550104 TI - High-dose etoposide and autologous bone marrow transplantation as intensification treatment in small cell lung cancer: a pilot study. AB - A pilot study was conducted in which 15 patients with small cell lung cancer (SCLC) with limited or extended disease were treated with high dose etoposide (600 mg/m2 daily for 3 consecutive days) followed by autologous bone marrow transplantation (ABMT). Twelve patients underwent a double graft. All had achieved complete or partial remission with conventional induction chemotherapy (adriamycin and etoposide, plus cisplatin in five cases). After ABMT six of the 15 patients did not receive radiotherapy to the chest; all but four patients received prophylactic brain irradiation. No toxic deaths were recorded during the period of aplasia. Eleven patients relapsed and died after ABMT. The median time to death was 18 months. One other patient died at 13 months of unknown cause. At the present time three patients are alive and free of disease at 54, 51 and 47 months respectively. This pilot study shows that high dose etoposide and ABMT is well tolerated as late intensification for responsive SCLC. Definite conclusions about its precise role in therapy cannot yet be drawn. PMID- 2550105 TI - Salivary cortisol in pigs following adrenocorticotrophic hormone stimulation: comparison with plasma levels. AB - Four prepubertal pigs were prepared with venous catheters and housed in metabolism cages. Plasma and saliva samples were taken at 15-min intervals over a 105-min period and analysed by radioimmunoassay for total (i.e. free and bound) cortisol content. Adrenocorticotrophic hormone (ACTH) was given i.v. at three different doses (0.5, 1.0 and 2.0 mg) after the second sample and the cortisol responses were compared with pretreatment values and levels observed after saline vehicle administration. Basal levels of salivary cortisol were approximately 10% of those in plasma. ACTH induced significant increases in plasma and salivary cortisol but in no case was a dose/response relationship detected. Plasma cortisol showed a maximum increase of approximately 230% whereas salivary cortisol increased only by about 130%, indicating that salivary cortisol is a less sensitive indicator of adrenal activity than plasma cortisol in this species. Estimation of salivary cortisol concentrations may offer practical advantages for the assessment of stress responses in intensively housed pigs. PMID- 2550106 TI - Guanine nucleotide regulation of [125I]beta-endorphin binding to NG108-15 and SK N-SH cell membranes: specific cation requirements. AB - Regulation of [125I]beta h-endorphin binding by guanine nucleotides was investigated in membrane preparations from two opioid receptor-containing cell lines: NG108-15, which contains only delta opioid receptors, and SK-N-SH, which contains predominantly mu opioid receptors. In contrast to the binding of the delta-selective agonist [3H][D-penicillamine2,D-penicillamine5]enkephalin to NG108-15 cell membranes, and of the mu-selective agonist [3H][D-Ala2,MePhe4,Gly ol5]enkephalin to SK-N-SH cell membranes, [125I]beta h-endorphin binding to NG108 15 and SK-N-SH cell membranes was not altered by guanosine triphosphate (GTP) or guanylyl-5'-imidodiphosphate (Gpp(NH)p) in the absence of cations. However, in the presence of NaCl, [125I]beta h-endorphin binding to both cell lines was inhibited by GTP and Gpp(NH)p in a concentration-dependent manner. In SK-N-SH cell membranes, the ability of sodium to promote regulation of [125I]beta h endorphin binding by GTP was mimicked by the monovalent cations lithium and potassium, but not by the divalent cations magnesium, calcium, or manganese. In NG108-15 cell membranes, only sodium was effective in promoting inhibition of [125I]beta h-endorphin binding by GTP. The effect of GTP or Gpp(NH)p in the presence of sodium was also observed with guanosine diphosphate, but not guanosine monophosphate or any of the non-guanine nucleotides tested. These results indicate that the presence of monovalent cations is required for regulation of [125I]beta h-endorphin binding by guanine nucleotides, and that the specificity of this cation requirement differs between the mu and delta receptor containing cell lines. PMID- 2550107 TI - ACTH-(1-24) enhances the electrically stimulated release of [3H]dopamine from rat septal slices via a dopamine D2 receptor-independent mechanism. AB - ACTH-(1-24) enhanced the basal as well as the electrically stimulated release of [3H]dopamine from rat septal slices in vitro. In the absence of Ca2+ from the superfusion medium the effect of ACTH-(1-24) on the electrically stimulated release of [3H]dopamine was abolished. The stimulus-evoked release of [3H]dopamine from septal slices appeared to be modulated through dopamine receptors of the D2 subtype: the dopamine D2 receptor agonists 2-(N-propyl-N-2 thienylethylamino)-5-hydroxytetralin (N-0437) and quinpirole reduced, whereas the dopamine D2 receptor antagonist sulpiride enhanced the electrically stimulated release of [3H]dopamine. The magnitude of the effect of ACTH-(1-24) on [3H]dopamine release was the same in the presence or absence of N-0437, quinpirole and sulpiride. ACTH-(1-24) had no effect on either the basal or the electrically stimulated release of [3H]noradrenaline. Also when the electrically stimulated release of [3H]noradrenaline was reduced by the alpha 2-adrenoceptor agonist clonidine, the peptide was without effect. These results show that ACTH (1-24) selectively enhances the release of [3H]dopamine from septal slices. The effect of the peptide is independent of the degree of activation of dopamine D2 receptors which modulate the stimulus-evoked release of [3H]dopamine. These results suggest that ACTH-(1-24) enhances the stimulus-evoked release of dopamine in the septum via a mechanism not associated with dopamine D2 autoreceptors. PMID- 2550108 TI - Noradrenergic innervation does not affect chronic regulation of [125I]pindolol receptors in fetal rat brain transplants or host neocortex. AB - Fetal (E15-16) somatosensory cortex (n = 15) or cerebellum (n = 9) were placed into the somatosensory cortex (SmI) of adult rat hosts to study the relative importance of tissue origin versus host milieu on graft beta-adrenoceptor regulation. Autoradiographic studies of [125I]pindolol ([125I]pin) binding in the presence of 3 microM serotonin were performed as an index of beta-receptor binding in both intact hosts and those with ipsilateral locus coeruleus (LC) lesions and/or ipsilateral superior cervical ganglionectomy. [125I]pin binding within fetal grafts was highly variable with areas of highest specific binding in cortical grafts (Kd = 209 +/- 30 pM, Bmax = 106 +/- 7 (fmol/mg protein) being comparable to host cortex (Kd = 211 +/- 41 pM, Bmax = 111 +/- 9 fmol/mg protein). Average total binding in whole cortical grafts was 73% and in cerebellar grafts was 60% of that in comparable adult cortex. Host cortex had 66-73% and cerebellum had 4-8% beta 1-receptors while cortical grafts had 59% and cerebellar grafts had 43% beta 1-receptors as determined by competitive binding with ICI 89406 and 118551. Noradrenergic fibers derived from both the host LC and superior cervical ganglion grew into fetal cortical grafts. Binding to high affinity uptake sites ([3H]desmethylimipramine, [3H]DMI) on noradrenergic terminals in cerebellar grafts was 28% higher than that in cortical grafts; superior cervical ganglionectomy decreased [3H]DMI binding in cortical grafts by 37% but had no effect on cerebellar grafts. Neither ganglionectomy nor LC lesions affected total specific binding or binding to beta-receptor subtypes in the grafts or host cortex 3-6 months after removal. Therefore, anatomic site of origin appeared to be the predominant factor in determining the development of beta-adrenoceptors in fetal cortical tissue. In ectopically placed cerebellar grafts, beta-receptor subtypes did not develop comparably to host cerebellar receptors suggesting that host milieu may be of critical importance in receptor development in this tissue. PMID- 2550109 TI - Different GTP-binding proteins mediate regulation of calcium channels by acetylcholine and noradrenaline in rat sympathetic neurons. AB - In dissociated neurons of rat superior cervical ganglion (SCG), noradrenaline (NA) and acetylcholine (ACh) suppressed Ca2+ currents elicited by depolarizations to 0 mV from -60 mV. With GTP-gamma-S in patch electrodes, ACh and NA caused persistent inhibition of Ca2+ currents. Pretreatment of SCG cells with pertussis toxin abolished the action of ACh but not of NA. The results suggest that ACh and NA reduce the Ca2+ currents in SCG cells through different G proteins. PMID- 2550110 TI - Subtypes of K+ channels differentiated by the effect of K+ channel openers upon K+ channel blocker-induced seizures. AB - Intracerebroventricular injection of mast-cell degranulating peptide (MCD), dendrotoxin I (DTXI) and 4-aminopyridine (4-AP), 3 blockers of a subclass of K+ channel, produces seizures and convulsions. Three different K+ channel openers are potent blockers of MCD-induced hyperexicitatory effects when they are administered preventively but they are unable to inhibit the epileptogenic effects induced by DTXI and 4-AP which were thought to block the same K+ channel which is blocked by MCD. PMID- 2550112 TI - Dark-induced changes in activity and compartmentalization of retinal calmodulin kinase in the rat. AB - Calmodulin kinase (CaM kinase) activity and immunoreactivity were measured in the cytosol and crude synaptic membranes of light- and dark-adapted rat retinas. Dark adaptation increased the calcium-independent CaM kinase activity 2.7 times and calcium-stimulated activity 3.9 times in membrane fractions. Dark adaptation also increased membrane-bound CaM kinase immunoreactivity 2.4 times. In the cytosol, dark adaptation increased calcium- and calmodulin-independent kinase activity 3.3 fold but did not enhance calcium- and calmodulin-dependent activity. Soluble CaM kinase immunoreactivity was decreased by 13% by dark exposure. These changes in enzyme activity and immunoreactivity are likely due to changes in the endogenous state of autophosphorylation and compartmental concentrations of CaM kinase and may represent translocation of CaM kinase from cytosol to membranes. CaM kinase may have an important role in modulating visual processes. PMID- 2550113 TI - [Clinical analysis of 106 surgically treated cases of stage I non-small cell lung cancer]. PMID- 2550111 TI - Analogies and differences in the mode of action and properties of binding sites (localization and mutual interactions) of two K+ channel toxins, MCD peptide and dendrotoxin I. AB - Both the bee venom toxin, mast cell degranulating (MCD) peptide, and the snake toxin, dendrotoxin 1 (DTX1) induce epileptiform activity and paroxystic seizures after intracerebroventricular (i.c.v.) injection to rats. Although many of the properties of the two toxins, which are blockers of the same K+ channel, appear to be very similar, a number of differences have been found. (1) Induced seizures have an hippocampal origin for MCD and two different origins, situated in the cortex and in the limbic system, for DTX1. (2) A first i.c.v. administration of DTXI desensitizes against a second ipsilateral injection of the same peptide as we had previously observed for MCD. However no cross-desensitization was observed between the two different toxins. (3) The number of high affinity (Kd = 41 pM) binding sites for 125I-DTXI in synaptic membranes is about 5 times higher than the number of high affinity (Kd = 158 pM) binding sites for 125I-MCD. (4) Autoradiographic analysis of the distribution of high affinity 125I-DTX1 binding sites has been compared to our previous analysis of high affinity 125I-MCD binding sites. High levels of high affinity binding sites for both toxins seem to be localized in synapse-rich areas. However high affinity binding sites for the two toxins are not always co-localized. Analysis of the mutual interactions between DTXI and MCD binding sites has revealed the presence of classes of low affinity binding sites for MCD. In most areas of the brain, a large proportion of high affinity binding sites for DTXI is allosterically related to low affinity binding for MCD. PMID- 2550114 TI - [Pathologic attributes and pathogenesis of brain edema in a rat model of cerebral ischemia with reperfusion]. PMID- 2550115 TI - [Changes of angiotensin II and angiotensin-converting enzyme in cor pulmonale with respiratory failure]. AB - Angiotensin II (ATII), angiotensin-converting enzyme (ACE) and plasma renin activity (PRA) were measured by radioimmunoassay and fluorometry in 55 cases of cor pulmonale with type II respiratory failure, 48 healthy people and 46 COPD patients. THE RESULTS: 1. The concentration of ATII in respiratory failure group was higher than that in the healthy people and in the COPD patients. The activity of ACE was lower and PRA was higher in the respiratory failure group than those in the other groups. 2. ATII in the dead was higher than that in the remittent. 3. After oxygen treatment, ATII of the patients with PaO2 greater than 8.0 kPa was lower than that of the patients with PaO2 less than 8.0 kPa, but ACE and PRA of the former were lower than those of the latter. 4. ATII and ACE of the respiratory failure group were closely related to PaO2. It was concluded that during hypoxemia one of the responses to high ATII in the human body was the decrease of ACE. But both the increase of PRA and the insufficient decrease of ACE in the patients with respiratory failure enhance the concentration of ATII. Also the concentration of ATII was related to the prognosis of the patients, and merely supplying the patient with oxygen cannot restore normal ATII. So, in addition to oxygen treatment, it is significant to inhibit PRA and ACE and use ATII as an indicator while dealing with cor pulmonale with cardiac and respiratory failure. PMID- 2550116 TI - [Study on human monoclonal antibodies in hemorrhagic fever with renal syndrome virus]. PMID- 2550117 TI - Effect of gallium on the in vitro formation, growth, and solubility of hydroxyapatite. AB - Gallium (Ga) is an effective treatment for the hypercalcemia of malignancy. The mechanism of action of the metal in blocking bone resorption in humans is not well understood. This paper examines the effect of Ga on the in vitro formation of hydroxyapatite (HA) in three test systems that have possible biological relevance in a pH-stat at pH 7.4, 37 degrees C, and 0.15 M NaCl: (1) the direct precipitation of HA; (2) the transformation of amorphous calcium phosphate to HA; and (3) the growth of HA seeds. In addition, the effect of Ga on HA solubility was measured at pH 5.0, the approximate pH of osteoclastic bone resorption. Ga decreased the HA formation and/or growth kinetics in a dose-related manner in all three test systems. In addition, the time to the onset of HA formation was increased in systems 1 and 2. Also, the adsorption of Ga on the surface of HA crystals was measured. Ga reduced the dissolution kinetics of HA compared with Ga free control. The mechanism reported herein--the significant adsorption of Ga on forming and growing HA nuclei and on the surface of HA crystals--is believed to be responsible for the effects of the metal on HA proliferation and solubility. Accumulation of the metal on newly formed metaphyseal bone can now be explained by this adsorption of Ga. These in vitro results partly explain the in vivo action of Ga in treating hypercalcemia by decreasing bone apatite solubility.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550118 TI - [Cytomegalovirus pneumonitis in renal transplantation--a case report]. AB - Cytomegalovirus pneumonitis is a severe complication of renal transplantation. It usually occurs in the first 3-4 months after transplantation. Herein, we describe a 21 year-old male patient with chronic renal failure who received a successful cadaveric renal transplantation after regular hemodialysis three times per week for 3 years. Cyclosporin A and Methylprednisone were used for immunosuppressants after operation. The allograft kidney function improved gradually in two weeks. Serum BUN and creatinine were lowered to 25 mg/dl and 1.7 mg/dl respectively. Bacterial pneumonia developed on the 16th postoperative day. The control of pneumonia rapidly improved after the use of broad spectrum antibiotics. However, bilateral diffuse pulmonary infiltrations developed 21 days later. He received transbronchoscopic diagnostic procedures including TBLB without definite diagnosis. The final diagnosis was obtained by open lung biopsy, in which intranuclear inclusion body was found. The patient expired on the 55th postoperative day despite adjustment of immunosuppresant dosage and use of antiviral therapy. PMID- 2550119 TI - ACTH receptors in nervous tissue. High affinity binding-sequestration of [125I]Phe2,Nle4]ACTH 1-24 in homogenates and slices from rat brain. AB - We have demonstrated specific, high affinity binding of a biologically active Tyr23-monoiodinated derivative of ACTH, [125I][Phe2,Nle4]ACTH 1-24, in rat brain homogenates. Similarly, in metabolically inhibited and noninhibited rat whole brain slices there is a specific "binding-sequestration" process that is dependent on time, protein concentration, and pH. In homogenates, binding curves were best described by a two-site model and provided the following parameters: Kd1 = 0.65 +/- 0.47 nM, Bmax1 = 21 +/- 41 fmol/mg protein; Kd2 = 97 +/- 48 nM, Bmax2 = 3.5 +/- 1.8 pmol/mg protein. In metabolically viable brain slices, concentration-competition curves of [125I][Phe2,Nle4]ACTH 1-24 binding sequestration can be described by three components (Kd1 = 14 +/- 24 nM, Bmax1 = 50 +/- 95 fmol/mg protein; Kd2 = 2.4 +/- 1.9 microM, Bmax2 = 44 +/- 49 pmol/mg protein; Kd3 = 0.16 +/- 1.0 mM, Bmax3 = 5.3 +/- 54 nmol/mg protein). Metabolic inhibition, by removal of glucose and addition of 100 microM ouabain, abolishes the lowest affinity, highest capacity binding-sequestrian component only (Kd1 = 7.1 +/- 14 nM, Bmax1 = 8.7 +/- 16 fmol/mg protein; Kd2 = 7.4 +/- 4.49 microM, Bmax2 = 37 +/- 27 pmol/mg protein). The two binding-sequestration parameter estimates obtained from metabolically inhibited tissue slices are not significantly different from those of the two higher affinity components obtained with noninhibited tissue. Thus, metabolic inhibition permits demonstration of ACTH receptor binding only, unconfounded by sequestration or internalization of ligand:receptor complexes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550120 TI - Ah receptor in spleen of rodent and primate species: detection by binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - In many species systemic toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is manifested by a generalized wasting syndrome accompanied by a variety of specific organ changes including atrophy of the thymus and spleen. TCDD toxicity in most tissues is thought to be mediated by the Ah receptor. Although the spleen is a prime target for TCDD toxicity, the possible presence of Ah receptor in the spleen has not previously been investigated. Specific binding of [3H]TCDD to Ah receptor in spleen cytosols was assessed by velocity sedimentation on sucrose gradients. Ah receptor was detected in spleen cytosols from adult Rhesus monkeys (mean +/- SEM, 36 +/- 8 fmol/mg cytosol protein), fetal Rhesus monkeys (9 +/- 6), Sprague-Dawley rats (20 +/- 5), C57BL/6J mice (18 +/- 2), New Zealand white rabbits (19 +/- 2), and Hartley guinea pigs (15 +/- 2). Ah receptor was not detectable in spleen cytosol from genetically "nonresponsive" DBA/2J mice or from Golden Syrian hamsters, a species resistant to toxicity of TCDD. Molecular properties of Ah receptor from spleen were similar to those of the receptor from liver of the same species. The high Ah receptor content in spleen cytosols from those species that are most susceptible to TCDD toxicity is consistent with the view that the Ah receptor mediates TCDD toxicity in spleen as well as in other tissues. PMID- 2550121 TI - Survival of patients with glioblastoma multiforme treated by intraoperative high activity cobalt 60 endocurietherapy. AB - The authors report their initial treatment results in 49 patients with glioblastoma multiforme (GM) who received intraoperative endocurietherapy (ECT) with high-activity cobalt 60 (60Co) probe. Thirty poor prognosis (unresectable tumor) patients (Group I) with newly diagnosed GM were treated by either biopsy or subtotal excision, followed by 20.00-Gy single-fraction 60Co probe ECT, and 60.00-Gy external-beam radiation therapy (EXRT) (80.00 Gy total tumor dose). Nineteen patients (Group II) with recurrent, previously resected and externally irradiated GM were retreated with 20.00-Gy single-fraction 60Co probe ECT alone. The authors' initial experience with intraoperative ECT of GM is discussed. PMID- 2550122 TI - Coexistence of cytoplasmic and nuclear estrogen receptors. A histochemical study on human mammary cancer and rabbit uterus. AB - Consecutive serial cryostat-frozen sections of 157 human mammary carcinomas and the uteri of six immature New Zealand white rabbits were stained histochemically for cytoplasmic estrogen receptor (ER) and nuclear ER by a fluorescent estrogen compound (Fluorocep Estrogen, Zeus Technologies, Inc., Raritan, NJ) and by a monoclonal antibody immunoperoxidase technique (ER-ICA, Abbott Laboratories, North Chicago, IL), respectively. The percentage of the ER-positive cells in the cancer cell population under observation was estimated and recorded. The results of the cytoplasmic ER assay were compared with those of the nuclear ER assay in each tumor; all cancers with less than 10% ER-positive cancer cells were grouped together as ER-negative tumors, the cancers with 30% or more ER-positive cancer cells as ER-positive tumors, and those with 10% to 29% ER-positive cancer cells as borderline positive. According to this manner of classification, 94% to 97% of the ER-positive mammary carcinomas diagnosed by one histochemical assay would have been identified as such by the other with no more than 10% difference in the ER-positive cell counts. The majority of ER-positive breast cancer cells and practically all of the luminal lining cells of the immature rabbit endometrium had coexistent cytoplasmic and nuclear ER. In the mammary cancers containing less than 30% ER-positive cancer cells, there was a greater (up to 20%) discrepancy in positive cell counts between the cytoplasmic ER assay and the nuclear ER assay. This discrepancy may be due to sampling errors of small clones of ER-positive cancer cells in two adjacent sections, difference in antigenic determinants between the cytoplasmic and the nuclear ER, and the binding sites in the nuclear ER being preoccupied by estrogen. The findings of this study appear to support the hypothesis that there are ER in the cytoplasm and the nucleus of the mammary carcinoma cells and the epithelial cells of the endometrium. PMID- 2550124 TI - Relation between DNA ploidy and the clinical behavior of phyllodes tumors. AB - We studied the DNA histograms obtained by flow cytometry from a series of six giant fibroadenomas and ten phyllodes tumors to determine if the analysis of DNA ploidy would help to predict clinical behavior. We were unable to document any relation between ploidy and histologic appearance, recurrence, metastasis, lesion size, or patient age. DNA aneuploid stem cell lines were seen in 75% of histologically benign phyllodes tumors, 50% of histologically malignant phyllodes tumors, and approximately 33% of giant fibroadenomas. PMID- 2550123 TI - Expression of ras oncogene p21 protein in early gastric carcinoma and adjacent gastric epithelia. AB - The expression of the ras gene product p21 in normal gastric mucosa, early gastric carcinoma of diffuse (gastric) and intestinal types, and in adjacent mucosal abnormalities is reported. The analysis was performed on paraffin sections by an immunohistochemical assay using the mouse monoclonal antibody RAP 5 and the rat monoclonal antibody Y13-259. Expression of ras p21 was assessed by staining intensity and percentage of positively stained cells. In comparison to normal gastric mucosa of non-cancer patients, p21 was overexpressed in nearly all early carcinomas of both types and in the dysplastic and/or metaplastic mucosal alterations accompanying intestinal type of gastric cancer. Increased p21 expression was also observed in the normal-appearing mucosa adjacent to early carcinomas of diffuse type, but not in the morphologically normal gastric epithelium adjacent to the intestinal type. The results of this investigation suggest that ras p21 overexpression may be related to early events of human gastric carcinogenesis. The study supports the notion of different pathways in the development of diffuse (gastric) and intestinal types of gastric carcinomas. PMID- 2550125 TI - Small cell carcinoma of the esophagus. The Memorial Hospital experience 1970 to 1987. AB - During the period 1970 to 1987, 11 patients with small cell carcinoma of the esophagus were treated at Memorial Sloan-Kettering Cancer Center, New York. This rare tumor was responsible for 1.1% of all patients with esophageal tumors seen on the inpatient services during that period. Using a clinical staging system similar to that employed in small cell cancer of the lung, eight of 11 patients had extensive disease. Although responses were seen to multidrug combination chemotherapy regimens used alone or with sequential radiation, the overall prognosis for small cell esophageal cancer was poor, with a median survival of 7.5 months. Only one patient lived for greater than 2 years. PMID- 2550126 TI - Enzymic synthesis of di- and tri-saccharide glycosides, using glycosidases and beta-D-galactoside 3-alpha-sialyl-transferase. AB - The following disaccharide glycosides were obtained in yields of 10-35% from the appropriate donor and acceptor glycosides by employing glycosidases as catalysts: alpha-D-Galp-(1----3)-alpha-D-GalpNAc-OEt (alpha-D-galactosidase), beta-D-Galp-(1 ---3)-alpha-D-GalpNAc-OEt and beta-D-Galp-(1----3)-beta-D-GalpNAc-OEtBr (beta-D galactosidase), beta-D-GlcpNAc-(1----6)-beta-D-Galp-OMe and beta-D-GlcpNAc-(1--- 6)-alpha-D-Manp-OMe (beta-N-acetylglucosaminidase). With beta-D-GlcpNAc-OEtSiMe3 as the acceptor, beta-D-galactosidase gave beta-D-Galp-(1----3)-beta-D-GlcpNAc OEtSiMe3 almost exclusively, whereas, with beta-D-GlcpNAc-OMe as the acceptor, beta-D-Galp-(1----3)-beta-D-GlcpNAc-OMe was formed in only slightly excess over teh analogous beta-(1----4)-linked glycoside. The use of beta-D-galactosidase and beta-D-galactoside 3-alpha-sialyltransferase in sequence provided a convenient route to the trisaccharide glycosides alpha-D-Neup5Ac-(2----3)-beta-D-Galp-(1--- 3)-alpha-D-GalpNAc-OEt, alpha-D-Neup5Ac-(2----3)-beta-D-Galp-(1----3)-beta-D GalpNAc-OE tBr, and alpha-D-Neup5Ac-(2----3)-beta-D-Galp-(1----3)-beta-D-GlcpNAc OMe. PMID- 2550127 TI - Proof of the occurrence of 5,6-O-(1-carboxyethylidene)-D-galactofuranose units in the capsular polysaccharide of Klebsiella K12. AB - The 13C-n.m.r. spectra of the capsular polysaccharide of Klebsiella K41 and phage derived oligosaccharides K41-P1 and K41-P2 were compared with spectra from the structurally similar polysaccharide of Klebsiella K12 and oligosaccharides K12-P1 and K12-P2. This led to the conclusion that K41 and K12 contain one and two galactofuranose residues per repeating unit, respectively, and that the terminal, lateral residue in K12 has the 5,6-O-(1-carboxyethylidene)-D-galactofuranose structure rather than that of a 4,6-acetal of D-galactopyranose as originally stated. This is the first reported occurrence in Nature of such a structural unit. PMID- 2550128 TI - (1----3)-alpha-D-glucan from an alkaline extract of Agrocybe cylindracea, and antitumor activity of its O-(carboxymethyl)ated derivatives. AB - The structure of an alkali-soluble D-glucan (AG-AL) from the fruit body of Agrocybe cylindracea was investigated by a combination of chemical and spectroscopic methods indicating that it was a linear (1----3)-alpha-D-glucan (molecular weight, approximately 560,000), [alpha]20D +195 degrees (c 0.5, M sodium hydroxide). Both water-soluble and gelatinous products obtained by O (carboxymethyl)ation of AG-AL showed potent antitumor activity against the solid form of Sarcoma 180 in mice, although the native D-glucan had little effect on the tumor. PMID- 2550129 TI - Superoxide production by human umbilical vein endothelial cells in an anoxia reoxygenation model. AB - Ischaemia/reperfusion of cardiac tissue has been claimed to be associated with the production of oxygen free radicals, which can contribute to severe membrane damage and tissue injury. We investigated the effects of anoxia/reoxygenation treatment on superoxide radical production in an in vitro system consisting of preconfluent and confluent human endothelial cell monolayers. The influence of varying the anoxia and reoxygenation phases on superoxide production was studied. As a test of cytotoxicity, the release of the cytosolic enzyme lactate dehydrogenase in the culture medium was measured before and at 0, 24 and 48 h after anoxia-reoxygenation. Cellular damage was monitored by microscopic examination of the cultures during and after the experiments and by the expression of the von Willebrand protein and of the membrane glycoprotein IIa by indirect immunofluorescence with specific monoclonal antibodies. Our results show that the endothelial cells subjected to anoxia-reoxygenation release superoxide anions, as revealed by superoxide dismutase inhibitable cytochrome C reduction. Free radical production is dependent on cell confluent or preconfluent state and on both anoxia and reoxygenation duration. Free radical release does not seem to be accompanied by manifest cellular alteration. PMID- 2550130 TI - Effects of chronic treatment with adrenaline or propranolol on platelet function and c-AMP levels in the rat. AB - Most of our knowledge about the modulation of platelet function by catecholamines is based on observations of acute in vitro actions. Little is known about the effects of chronically elevated or reduced adrenergic stimulation of the platelets. We therefore treated rats for 8 weeks with either adrenaline or the beta-blocker propranolol. Adrenaline (0.5 mg.kg-1.d-1) continuously administered from subcutaneously implanted osmotic mini pumps caused an increase in the sensitivity of the platelets towards ADP as stimulating agent. In contrast, chronic application of propranolol (10 mg.kg-1.d-1) via the drinking water led to a reduction in platelet aggregability. For animals treated with adrenaline, in accordance with the results of the aggregation experiments, the levels of c-AMP found in platelet rich plasma were reduced, both basally (by 33%) and after stimulation of platelet adenylate cyclase with prostaglandin E1 (by 39%). For the propranolol treated animals, the basal c-AMP concentrations remained unchanged. The levels of c-AMP attained after stimulation with prostaglandin E1 were diminished to a similar extent as for the adrenaline treated animals (by 38%). Although the in vitro addition of adrenaline to platelet rich plasma causes a beta-adrenoceptor mediated inhibition of platelet aggregation in the rat, the simulation seen after chronic adrenaline exposure in vivo, which is associated with decreases in both basal and stimulated c-AMP levels, suggests a functional preponderance of alpha-adrenoceptors over beta-adrenoceptors on the rat platelets. Although intraplatelet metabolic changes (blockade of stimulated c-AMP formation) after chronic application of propranolol should have resulted in enhancement of platelet aggregability, an inhibition of aggregation was observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550131 TI - Electrical activity of human atrial fibres at frequencies corresponding to atrial flutter. AB - Little information is available about rate dependent changes in electrical activity of human myocardial cells. We therefore studied, in vitro, the electrical activity of adult human atrial fibres driven at frequencies near that of atrial flutter by means of the standard microelectrode technique. Thirty two atrial samples exhibiting "normal" responses with fast upstroke were selected. At very high frequencies, the action potential (AP) upstroke arose from the repolarisation phase of the preceding AP in spite of marked frequency induced shortening of the plateau. As the stimulation rate was progressively increased, the take off potential (TOP) was less and less negative and the maximal rate of depolarisation (Vmax) decreased. Moreover, in most preparations, a clear alternation between two types of action potentials occurred. Calcium channel inhibitors cobalt (5 mM) or diltiazem (5 x 10(-6) M) shortened AP duration, increased Vmax and markedly reduced alternation. Sodium channel inhibitors, tetrodotoxin (7.5 10(-6) M) or lignocaine (10(-5) M) shortened AP duration and induced a transient increase in Vmax. Ouabain (10(-6) M) prolonged AP duration, decreased Vmax, enhanced alternation and finally suppressed the 1:1 capture of the atrial tissue. Our results show that, at high driving rates corresponding to the frequencies of atrial flutter, slight variations in action potential duration induced by drugs are associated with marked concomitant variations in Vmax and probably with consequent modifications of the conduction velocity. PMID- 2550132 TI - The anti-arrhythmic effects of myocardial ischaemia. Relation to reperfusion arrhythmias? AB - Ventricular tachycardia was induced in the intact non-ischaemic pig heart by intramyocardial or intracoronary infusions of noradrenaline or N6, O2'-dibutyryl cAMP. The chemically induced tachycardia was consistently stopped within 10 to 30 s by occluding the coronary artery supplying the infusion area. This ischaemic effect was readily reversed by coronary reperfusion, with ventricular tachycardia resuming within seconds after release of the occlusion. In contrast to the immediate effect of myocardial ischaemia, it took several minutes for the tachycardia to cease after the infusion of arrhythmogenic compounds was stopped. Pacing experiments showed that the effect of myocardial ischaemia on ventricular tachycardia was probably not due to a conduction block. The anti-arrhythmic property of myocardial ischaemia was separate from its known effect of decreasing the ventricular fibrillation threshold for electrical stimulation. The increased vulnerability of the acutely ischaemic myocardium to fibrillation was apparent in experiments in which ectopic activity was induced in the non-ischaemic part of the myocardium. In these experiments ventricular fibrillation consistently ensued within 6 min following distal occlusion of the anterior descending coronary artery. By contrast, ventricular fibrillation was not precipitated by coronary artery occlusion or local infusion of arrhythmogenic compounds alone. Cyclic AMP was shown to accumulate in ischaemic myocardium. An association existed between cAMP accumulation and the intensity of early ischaemic arrhythmias as well as reperfusion arrhythmias. The highest incidence of ventricular fibrillation was found during reperfusion, at peak myocardial cAMP levels. These findings suggest: (1) Noradrenaline and dibutyryl-cAMP exert arrhythmogenic effects preferentially in the intact, non-ischaemic myocardium, the effects being attenuated in ischaemic myocardium by a paradoxical anti-arrhythmic effect of ischaemia. (2) In the acutely ischaemic heart, ventricular fibrillation may be precipitated by the emergence of ectopic activity outside the ischaemic area. (3) Arrhythmias and fibrillation occurring early after reperfusion may be caused by unmasking the effects of excitants (eg, noradrenaline or cAMP) arising during the antecedent period of ischaemia. PMID- 2550134 TI - Prophylactic use of hyperimmunoglobulin for cytomegalovirus infection in heart transplantation. AB - Cytomegalovirus (CMV) infection in solid organ recipients can endanger the immunosuppressed patient and increase vulnerability to secondary infections and the high risk of rejection triggered by the viral disease. The effect of passive immunization against CMV was examined in 69 heart transplant patients. The patients received weekly administrations of 1 ml/kg of CMV hyperimmunoglobulin from the day of transplantation until the 30th postoperative day. Forty-four of the patients were monitored clinically and serologically up to the 120th postoperative day. Nine patients showed clinical and serologic signs of CMV infection; in 15 the only evidence of CMV infection was a rise in antibody titers. The remaining 20 patients showed no clinical or serologic signs of CMV infection. Three patients who were seronegative preoperatively remained seronegative until the end of the observation period. The results indicate a potential therapeutic benefit of hyperimmunoglobulin prophylaxis to prevent infectious complications due to CMV in heart transplant patients. PMID- 2550133 TI - Effects of the angiotensin converting enzyme inhibitors captopril, rentiapril, and alacepril in patients with essential and renovascular hypertension. AB - The effects of captopril, rentiapril, and alacepril were compared in five patients with renovascular hypertension (RVH) and five with essential hypertension (EH). The dose of each drug was equivalent to 50 mg of captopril. Rentiapril inhibited angiotensin converting enzyme activity more strongly and for longer periods than did the other two drugs. In the patients with RVH, blood pressure was significantly reduced more quickly and for longer periods by rentiapril than by captopril or alacepril; in the patients with EH, alacepril was the most potent antihypertensive agent. The results indicate that rentiapril is as effective as the other two drugs in patients with hypertension and that alacepril is more effective in patients with low-renin hypertension or renin independent hypertension than captopril or rentiapril. PMID- 2550135 TI - Treatment of malignant hypertension with an angiotensin converting enzyme inhibitor. AB - Malignant hypertension developed in an 18-year-old man whose primary hypertension had been diagnosed by chance. Standing blood pressure was 290/170 mmHg. Tests of renal function revealed high blood urea nitrogen and creatinine levels and low levels of both effective renal plasma flow and the glomerular filtration rate. Plasma renin activity and levels of angiotensin II and aldosterone were greatly elevated. Severe concentric left ventricular hypertrophy was noted. The patient received standard antihypertensive treatment with furosemide, propranolol, nifedipine, and prazosin, but his blood pressure did not decrease and there was no improvement in the clinical or biochemical measures. The patient was then given 20 mg of enalapril daily for one year. The inhibition of angiotensin converting enzyme immediately reduced blood pressure. Angiotensin II and aldosterone levels became normal, kidney function and hemodynamics improved, and echocardiograms revealed that the left ventricular hypertrophy had regressed. The results confirm the pathogenetic role of angiotensin II in the development of the malignant phase of hypertension. PMID- 2550136 TI - Cyclic AMP raises cytoplasmic calcium in pancreatic alpha 2-cells by mobilizing calcium incorporated in response to glucose. AB - The cytoplasmic Ca2+ concentration ([Ca2+]i) was monitored in individual guinea pig pancreatic alpha 2-cells exposed to modulators of glucagon release. Addition of the stimulatory amino acid arginine resulted in a sustained increase in [Ca2+]i, whereas the inhibitor glucose had the opposite effect. Epinephrine, the beta-adrenergic agonist isoproterenol, the adenylate cyclase activator forskolin and 8-bromo-cAMP transiently raised [Ca2+]i provided that the cells had been pretreated with glucose. However, simultaneous presence of glucose was not required and the effect occurred even in the absence of extracellular Ca2+. Carbachol, the alpha 2-adrenergic agonist clonidine and the sulfonylurea tolbutamide lacked effects on [Ca2+]i. In addition to providing support for the concept that glucagon release is positively modulated by [Ca2+]i, the results demonstrate that cAMP raises [Ca2+]i in the alpha 2-cells by mobilizing calcium incorporated in response to glucose. PMID- 2550137 TI - Alterations in binding of inositol 1,4,5-trisphosphate to subcellular structures of rat liver during chronic endotoxemia. AB - Inositol 1,4,5-trisphosphate (IP3) binding to, and Ca2+ uptake and release by plasma membrane- and endoplasmic reticulum-enriched fractions of rat liver were measured after continuous Escherichia coli endotoxin (ET) administration in vivo. IP3 binding to both fractions was significantly reduced by ET treatment. This was associated with decreased Ca2+ uptake and impaired IP3-dependent Ca2+ release. A decrease of 5'-nucleotidase specific activity of plasma membrane-enriched fraction was also observed in ET treated rats. The results suggest that previously observed impairments in the ability of hepatocytes to mobilize Ca2+, to activate glycogen phosphorylase and to respond--when saponin permeabilized--by Ca2+ release upon IP3 addition during chronic endotoxemia are due to alterations in both IP3 binding to the subcellular fractions that are imputed to be targets of IP3, and a decrease in the size of IP3-sensitive pool of releasable Ca2+. PMID- 2550138 TI - MyoD is a sequence-specific DNA binding protein requiring a region of myc homology to bind to the muscle creatine kinase enhancer. AB - MyoD is a skeletal muscle-specific protein that is able to induce myogenesis in a wide variety of cell types. In this report, we show that MyoD is a DNA binding protein capable of specific interaction with two regions of the mouse muscle creatine kinase gene upstream enhancer, both of which are required for full muscle-specific enhancer activity. MyoD shares antigenicity and DNA binding specificity with MEF1, a myocyte-specific DNA binding factor. The contiguous basic and myc homology regions of MyoD that are necessary and sufficient for specific DNA interaction are the same regions of the protein required to convert 10T1/2 fibroblasts into muscle. These findings suggest that the biological activity of MyoD is mediated via its capacity for specific DNA interaction. PMID- 2550139 TI - Temporal fluctuations in HIV quasispecies in vivo are not reflected by sequential HIV isolations. AB - A genetic study has been made of the HIV tat gene from sequential HIV-1 isolates and the corresponding infected peripheral blood mononuclear cells. DNA was amplified by polymerase chain reaction (PCR) and cloned into a eukaryotic expression vector. Twenty clones were sequenced from each sample. Comparing the sequential HIV isolates, abrupt differences were seen between the major forms of each isolate. These progressive changes were not reflected at all among the in vitro samples. The fluctuation in the quasispecies in vivo may suggest a much more dynamic role for latently infected mononuclear cells. High frequencies of functionally defective tat genes were identified. Given such complexity and the evident differences between quasispecies in vivo and in vitro, the task of defining HIV infection in molecular terms will be difficult. PMID- 2550140 TI - Protein-tyrosine phosphatases: the other side of the coin. PMID- 2550141 TI - Cytosolic protein translocation factors. Is SRP still unique? PMID- 2550142 TI - Erythropoietin receptor and interleukin-2 receptor beta chain: a new receptor family. PMID- 2550143 TI - Evidence that the leukocyte-common antigen is required for antigen-induced T lymphocyte proliferation. AB - The leukocyte-common antigen (L-CA) is a family of large molecular weight glycoproteins uniquely expressed on the surface of all nucleated cells of hematopoietic origin. The glycoprotein consists of a heavily glycosylated exterior domain, a single membrane spanning region, and a large cytoplasmic domain that contains tyrosine phosphatase activity. To investigate the function of this family, we generated T cell clones that lacked L-CA (L-CA-). The expression of the alpha beta T cell receptor, CD3, CD4, IL-2 receptor (p55), LFA 1, Thy-1, and Pgp-1 (CD44) was normal. The L-CA- T cell clones failed to proliferate in response to antigen or cross-linked CD3; however, they could still proliferate in response to IL-2. An L-CA+ revertant was obtained and the ability to proliferate in response to antigen and cross-linked CD3 was restored. These data indicate that L-CA is required for T cells to enter into cell cycle in response to antigen. PMID- 2550144 TI - Autophosphorylation of the PDGF receptor in the kinase insert region regulates interactions with cell proteins. AB - We have identified two platelet-derived growth factor (PDGF)-dependent autophosphorylation sites in the beta subunit of the human PDGF receptor (PDGF R). The major site of phosphorylation (Tyr-857) corresponds to the major autophosphorylation site in many other tyrosine kinases. Tyr-751, which lies within the kinase insert region, is a second in vivo site and the major in vitro site. Immunoprecipitates of wild-type PDGF-Rs prepared from PDGF-treated cells contained a phosphatidylinositol (PI) 3 kinase activity and three specific polypeptides as well as the PDGF-R. Mutation of Tyr-751 to Phe or Gly, or mutation of the catalytic domain to abolish kinase activity, blocked association of the PDGF-R with the PI kinase and the three proteins. These results suggest that autophosphorylation in the kinase insert region triggers the binding of the activated PDGF-R to specific cellular proteins, including a PI kinase whose activity is known to be stimulated by PDGF. Thus autophosphorylation may play a novel role in signal transduction via the PDGF-R. PMID- 2550145 TI - Molecular analysis of the para locus, a sodium channel gene in Drosophila. AB - Previous behavioral, electrophysiological, pharmacological, and genetic studies of mutations of the para locus in Drosophila melanogaster suggested that these mutations alter the structure or function of sodium channels. To identify the protein encoded by this gene and to elucidate the molecular basis of the mutant phenotypes, genomic DNA from the para locus was cloned. Mutational lesions in nine different para alleles were mapped and found to be distributed over a region of 45 kb. Analysis of cDNAs revealed that the para locus comprises a minimum of 26 exons distributed over more than 60 kb of genomic DNA. The nucleotide sequence of the complementary DNA predicts a protein whose structure and amino acid sequence are extremely similar to those of vertebrate sodium channels. The results support the conclusion that para encodes a functionally predominant class of sodium channels in Drosophila neurons. Furthermore, the para transcript appears to undergo alternative splicing to produce several distinct subtypes of this channel. PMID- 2550146 TI - Isolation of the gene encoding the yeast TATA binding protein TFIID: a gene identical to the SPT15 suppressor of Ty element insertions. AB - We report the cloning of the gene that encodes the yeast TATA binding protein TFIID. TFIID contains 240 amino acids and has no obvious sequence similarity to other known proteins. TFIID was synthesized in vitro and in two separate assays behaved identically to the protein purified from yeast. TFIID bound to TATA elements from the adenovirus major late promoter (TATAAAA) and the yeast LEU2 promoter (TATTTAA) and formed protein-DNA complexes stable to electrophoresis only in the presence of TFIIA. In vitro-synthesized yeast TFIID also complemented a mammalian in vitro transcription system that lacked TFIID. Comparison of the yeast TFIID gene with the yeast SPT15 gene (suppressor of Ty element insertions) showed that the two genes are identical. This finding indicates that the yeast TFIID activity defined in vitro is responsible for specific transcription in vivo. PMID- 2550147 TI - Lysis of P3HR-1 cells induced to enter the viral cycle by antibody-dependent and independent immunological mechanisms. AB - The P3HR-1 Burkitt lymphoma line carries the Epstein-Barr virus (EBV) genome and a small proportion of the cells (1-3%) enter the lytic cycle spontaneously. Treatment with TPA and n-butyrate elevates considerably the number of virus producing cells (25-35%). Cells which enter the lytic cycle express the EBV early antigen EA, the viral capsid antigen VCA, and the membrane antigen MA. Antibodies against these antigens are present in EBV-immune human sera. The expression of virus envelope protein on the plasma membrane renders the cells sensitive to immune effector mechanisms. These were shown to be initiated by the alternative complement pathway (ACP)-activating capacity of the cells and by their reactivity with antibodies directed to the MA. When incubated with EBV-immune or nonimmune human serum, the induced (P3HR-1-V) cells activated C3 through ACP and fixed the generated C3 fragments. The efficiency of opsonization was higher in immune serum. By varying the experimental conditions we showed the damage of the induced cells by the complement system and by blood lymphocytes, and analysed the involvement of antibodies and the activated C3 fragments in the lymphocyte mediated lysis. P3HR-1-V cells were lysed by immune serum and also by nonimmune serum though with lower efficiency. The induced cells had elevated sensitivity to the NK effect which was potentiated if the conditions allowed their opsonization. In the presence of antibodies the lymphocyte-mediated lysis was considerably higher and the ADCC mechanism was also potentiated by opsonization. These experiments suggest that B cells which enter the virus-producing cycle may be eliminated in EBV nonimmune host by NK cells. After the antibody response against the virus develops, the attack on these cells is more efficient through complement and lymphocyte-mediated antibody-dependent mechanisms. These effector mechanisms are enhanced by opsonization which is the consequence of the C3 activating capacity of the cells. The multiple ways of the immune attack on the B cells prepared to produce EBV may explain the absence of EA and VCA positive B cells in tumor cell populations and during the acute phase of infectious mononucleosis. PMID- 2550148 TI - Alteration of in vitro murine peritoneal macrophage function by dietary enrichment with eicosapentaenoic and docosahexaenoic acids in menhaden fish oil. AB - The effects of diets containing menhaden fish oil (MFO), compared with those of diets containing safflower oil (SAF) or an essential fatty acid deficient hydrogenated coconut oil (HCO), on in vitro activation of tumoricidal capacity by murine macrophages were assessed. Mice fed the experimental diets for 4 weeks were injected intraperitoneally with sterile thioglycollate broth 3 days before use. There was no difference between any of the groups with respect to total peritoneal exudate cells or the percentage of macrophages, although the fatty acid profile of purified adherent macrophages closely paralleled that of the diets. Macrophages from mice fed MFO killed fewer P815 mastocytoma cells upon activation with recombinant interferon gamma (IFN gamma) and lipopolysaccharide. Macrophages from all diets were equally competent for tumoricidal capacity when activated pharmacologically with calcium ionophore, phorbol 12-myristate 13 acetate, and lipopolysaccharide (LPS), suggesting that MFO diet macrophages were hyporesponsive to IFN gamma. Priming with higher concentrations of IFN gamma restored the partial defect in activation of MFO macrophages. When activated for 24 hr with high levels of LPS, macrophages from mice fed SAF displayed little cytolytic capacity; addition of indomethacin. (1 microM) resulted in enhanced levels of P815 kill. In contrast, MFO and HCO diet macrophages were highly cytolytic with similar LPS treatment with or without indomethacin. Macrophages from mice fed SAF produced threefold more prostaglandin E in response to LPS than did MFO and HCO diet macrophages. These results suggest that dietary manipulation of fatty acids can alter activation of tumoricidal capacity of macrophages, possibly both dependent and independent of changes in eicosanoid synthesis. PMID- 2550149 TI - Chemoattractant-elicited translocation of myosin in motile Dictyostelium. AB - The distribution of myosin was studied in amebae of the Ax-3 and NC-4 strains of Dictyostelium migrating at room temperature, using indirect immunofluorescence of aggregation-competent amebae and the agar-overlay technique. Amebae were fixed in methanol-formaldehyde or absolute acetone at -15 degrees C before or after stimulation with micromolar cyclic AMP at room temperature (20-25 degrees C). Myosin was detected by monoclonal antibodies to Dictyostelium myosin heavy chain followed by a fluorescent secondary antibody that had been preabsorbed to remove nonspecific staining. In both strains there was a striking increase in intensity of anti-myosin immunofluorescence in the cortex where it appeared as a continuous ring 30 seconds after addition of cyclic AMP. This correlated with a rounding up of the cell body. Sixty seconds after stimulation there was a clear reduction of cytoplasmic myosin rods in conjunction with the increased cortical localization. At this time extensions of largely hyaline cytoplasm were observed that extended beyond the cortical shell of myosin. Two minutes after the stimulus the immunofluorescence remained as a distinct line at the cortex, but the cells began to resume in elongated shape. By 3 minutes (NC-4 strain) or 5 minutes (Ax-3 strain) the amebae had largely returned to the control shape, and myosin had returned to its control distribution. Counts of the treated cells at different time points substantiated the observations of individual cells. The time course of translocation of myosin in the Ax-3 strain parallels the time course of myosin phosphorylation reported in previous studies. The results are interpreted in terms of a working hypothesis for the mechanism of translocation. PMID- 2550150 TI - Molecular transformation of Fusarium solani with an antibiotic resistance marker having no fungal DNA homology. AB - A vector was constructed for transformation of the plant pathogenic fungus Fusarium solani. The promoter 35Sp, from cauliflower mosaic virus, was fused to the bacterial gene APH(3')II, which confers resistance to the aminoglycoside antibiotic G418. Two transformation procedures were developed: one using isolated fungal protoplasts, the other using germinated fungal spores. A transformation frequency of 3.3 G418-resistant colonies were obtained per microgram DNA. Of 14 colonies analyzed, 12 had vector sequences integrated into their high molecular weight DNA, and 2 were untransformed. Integration was sometimes accompanied by rearrangements of both the vector and flanking fungal DNAs. Primer-extension analysis of the mRNA from one transformant revealed two putative transcription initiation sites in the chimeric APH(3')II gene. Both sites differed from the normal initiation site in plants. This vector will be useful in transformation systems in which integration by non-homologous recombination is desired. PMID- 2550151 TI - The role of iron in oxygen radical mediated lipid peroxidation. AB - The role of iron in the peroxidation of polyunsaturated fatty acids is reviewed, especially with respect to the involvement of oxygen radicals. The hydroxyl radical can be generated by a superoxide-driven Haber-Weiss reaction or by Fenton's reaction; and the hydroxyl radical can initiate lipid peroxidation. However, lipid peroxidation is frequently insensitive to hydroxyl radical scavengers or superoxide dismutase. We propose that the hydroxyl radical may not be involved in the peroxidation of membrane lipids, but instead lipid peroxidation requires both Fe2+ and Fe3+. The inability of superoxide dismutase to affect lipid peroxidation can be explained by the fact that the direct reduction of iron can occur, exemplified by rat liver microsomal NADPH-dependent lipid peroxidation. Catalase can be stimulatory, inhibitory or without affect because H2O2 may oxidize some Fe2+ to form the required Fe3+, or, alternatively, excess H2O2 may inhibit by excessive oxidation of the Fe2+. In an analogous manner reductants can form the initiating complex by reduction of Fe3+, but complete reduction would inhibit lipid peroxidation. All of these redox reactions would be influenced by iron chelation. PMID- 2550152 TI - DNA damage, cytotoxicity and free radical formation by mitomycin C in human cells. AB - Mitomycin C (MMC), a quinone-containing antitumor drug, has been shown to alkylate DNA and to form DNA cross-links. The ability of MMC to alkylate O6 guanine and to form interstrand cross-links (ISC) has been studied using Mer+ and Mer- human embryonic cells. Mer+ (IMR-90) cells have been reported to contain an O6-alkylguanine transferase enzyme and are, in general, more resistant to alkylating agents than the Mer- (VA-13) cell line, which is deficient in the repair of O6-lesions in DNA. Studies reported here show that MMC is more cytotoxic to VA-13 cells compared to IMR-90 cells. The alkaline elution technique was used to quantify MMC-induced ISC, and double strand breaks (DSB) in these cells. The drug-dependent formation of DSB was significantly lower in IMR-90 cells than in VA-13 cells. In contrast, no significant difference in cross linking could be detected at the end of 2-h drug treatment. Although a small increase in cross-link frequency was observed in the VA-13 cell line relative to the IMR-90 cell line 6 h post drug treatment, it is not clear whether monoalkylated adducts at the O6-position are formed, and contribute to cross-link formation for differential cytotoxicity in VA-13 cells. Electron spin resonance and spin-trapping technique were used to detect the formation of hydroxyl radical from MMC-treated cells. Our studies show that MMC significantly stimulated the formation of hydroxyl radical in VA-13 cells, but not in the IMR-90 cells. The formation of the hydroxyl radical was inhibited by superoxide dismutase (SOD) and catalase. In addition, the presence of these enzymes partially protected VA-13 cells from MMC toxicity but not IMR-90 cells. Further studies indicated that the decreased free radical formation and resistance to MMC may be due to the increased activities of catalase and glutathione transferase in the IMR-90 cell line. These results suggest that MMC-dependent DNA damage (alkylation and DNA DSB) and the stimulation of oxy-radical formation may play critical roles in the determination of MMC-induced cell killing. PMID- 2550153 TI - Amphotericin B-induced damage of Trypanosoma cruzi epimastigotes. AB - Amphotericin B (AmB) autoxidation resulted in oxygen consumption, superoxide anion formation and production of thiobarbituric acid (TBA)-reactive material (malondialdehyde). Malondialdehyde formation increased after incubation of the drug with ascorbate-ADP-FeCl3. Growth of Trypanosoma cruzi epimastigotes in the presence of AmB induced a decrease in the free fatty acid content of the cells (57% in control cells vs. 7% in AmB-treated cells), and in the proportion of unsaturated fatty acids as well as cell killing. No changes were detected on sterol content. No evidence was found for lipid peroxidation as a mechanism of cell injury by this antibiotic. PMID- 2550154 TI - Studies on proton pump inhibitors. I. Synthesis of 8-[(2-benzimidazolyl)sulfinyl] 5,6,7,8-tetrahydroquinolines and related compounds. AB - Many 8-[(2-benzimidazolyl)sulfinyl]-5,6,7,8-tetrahydroquinolines were synthesized and examined for their (H+ + K+) adenosine triphosphatase ATPase-inhibitory and antisecretory activities. These sulfinyl compounds could be considered to be rigid analogues of the 2-[(2-pyridyl)methylsulfinyl]benzimidazole class of antisecretory agents. All the compounds tested were potent inhibitors of (H+ + K+)ATPase. Most of the compounds also inhibited histamine-induced gastric acid secretion in rats. Among them, 8-[(5-fluoro-2-benzimidazolyl)sulfinyl]-3-methyl 5,6,7,8-tetrahydroqu inoline (XIVm) was found to have the most potent activity. The structure-activity relationships are discussed. PMID- 2550155 TI - [Establishment of a transplantable human rectal mucoid adenocarcinoma model in nude mice and study of its biological characteristics]. AB - A model of transplantable human mucoid adenocarcinoma of rectum in BALB/C nu/nu nude mice (TNB 92), was established and 18 sub-transplantations were performed. The success rate of transplantation was 98% and the average tumor bearing life time of mice was 112 days. Histology and ultrastructures showed that the transplantable tumor retained the original structures of the human tumor. Chromosomal analysis of the tumor cells exhibited the same features of the human carcinoma and it also retained the function of secreting CEA. Frozen tissue of the tumor in liquid nitrogen after rehabilitation could be successfully retransplanted into nude mice again. It seems to be a useful model for further study of human rectal adenocarcinoma. PMID- 2550156 TI - [An immunohistological study of monoclonal antibody BG6 against human breast cancer]. AB - Paraffin sections from a series of human tissues were examined by an indirect immunoperoxidase method with BG6 antibody. Data showed that primary breast cancer (96%) or lymph nodes with metastatic tumors (94%) gave positive result with BG6 antibody. Weak staining was observed in some sections of nonbreast tumors and no evidence of positive immunoperoxidase staining was obtained in benign breast hyperplasia, normal tissues and embryonic tissues examined. It is considered that McAb BG6 might be used as a specific marker for breast cancer. PMID- 2550157 TI - [Solubilization and purification of opioid receptor molecules of rat brain]. AB - P2 membrane preparation of rat brain (without cerebellum) was solubilized with CHAPS in Tris containing DTT and trypsin inhibitor. Two opiate ligands, 10b and 10cd, prepared by Liu et al, were applied consecutively in affinity chromatography, from which OPRs were eluted with Nx. The eluate was subsequently passed through a WGA affinity column and the OPR eluted with N-GluNAc. This eluate was further purified and concentrated by preparative granular gel isoelectric focusing on SG200. Two protein peaks appeared separately at pH 5 and pH7.8. The eluates from both peaks were analyzed for protein content by using silver staining method and binding activity was measured by RRA with 3H-etor. The results revealed that both samples contained active OPR purified to over 80,000 fold. The Mr was estimated by gel filtration to be 52 and 42 kD for OPR in the pH 5 and pH 7.8 samples respectively. The OPR in pH 5 sample has been characterized to be of mu-type, by its binding activity with 3H-ohm. PMID- 2550158 TI - [The relation of typing of gastric carcinoma according to traditional Chinese medicine theories and clinical pathologic classification]. AB - 102 cases of gastric carcinoma which have been removed by partial gastrectomy and have pathological diagnoses and follow-up results are reported. The relationship between their preoperational syndromes of TCM and pathological, TNM classification and post-operational survival rate has been studied. There is indication that the type of Ganwei Buhe (the dispersing function of the liver is disturbed and affects the stomach) manifests the earlier stage of the gastric carcinoma and the majority of cases are in stage 1 or 2 in TNM, the 5-year survival rate with surgery being 46%. The majority of cases of the types of Pishen Yangxu (deficiency of energy in the Spleen and Kidney), Yinxu Neire (the febrile syndrome of the viscera caused by the insufficient primordial energy) and Qixue Shuangkui (deficiency of both energy and blood) are in stage 3 or 4 in TNM, the 5-year survival rate with surgery differently being 5,8,12% respectively. The type of Qizhi Xueyu (energy stagnancy and blood stasis) is between the abovementioned two groups, with a 5-year survival rate of 40%. PMID- 2550159 TI - [Preliminary study of traditional Chinese medicine-Western medicine treatment of patients with primary liver carcinoma]. AB - In order to improve the therapeutic efficacy for patients with primary liver carcinoma (PLC), the authors treated 30 patients by routine chemotherapy in combination with the immunostimulators Bai Nian Le ( ), levamisole and cimetidine. As a result, the NK activity and percentage of lymphoblast transformation of patients were significantly elevated, expansion of the tumor mass was checked, with clinical conditions obviously improved. So the short term effect of the therapy was satisfactory. The study suggested that Bai Nian Le in combination with levamisole and cimetidine was able to elevate the immune response and therapeutic effect. It may be one of the useful adjuvant therapy for PLC patients. PMID- 2550160 TI - [Effect of Astragalus membranaceus on electrical activities of coxsackie B-2 virus-infected rat myocardial cells in culture]. AB - Beating Myocardial cell cultures of neonatal rats were prepared in vitro and infected with coxsackie B-2 virus. The cells were evaluated in the post-infected period for changes in beating percentage and cytopathic effect (CPE), alterations in the electrical activities by standard microelectrode techniques, and the protective effect of Astragalus membranaceus (AM) on coxsackie B-2 virus-infected neonatal rat myocardial cell cultures was observed. The beating percentage began to decrease in the infected group at 24 hr and only 27.9 +/- 18.6% was beating at 96 hr after virus challenge, premature beats, tachycardia and fibrillation occurred commonly during the experiment. Meanwhile the CPE appeared rapidly from 1+-3+ at the same interval. Resting potential, action potential amplitude, duration and rate of uptake were shown a significant decrease through 24-96 hr (P less than 0.01). In contrast, the beating and electrical activities were nearly normal and less CPE was shown in myocardial cells treated with AM 1 hr after virus challenge through 24-96 hr (P less than 0.05). The results suggest that AM may be valuable in prophylaxis and treatment of acute coxsackie B-2 virus caused myocarditis. PMID- 2550161 TI - NGF receptor increase in the olfactory bulb of the rat after early odor deprivation. AB - In the olfactory bulb of normal rats, nerve growth factor (NGF) receptor (NGFR) immunoreactivity was largely confined to the glomerular layer. Unilateral closure of the nostril at postnatal day 2 (P2) increased NGFR immunoreactivity in the sealed bulb at both 19 and 60 days after the operation. The increase in NGFR density, measured by autoradiographic immunohistochemistry, was most dramatic 60 days postocclusion. These findings suggest that a compensatory increase in NGFRs may play a role in the maintenance of bulbar function after the early loss of sensory stimulation. PMID- 2550162 TI - Lipids as endogenous Na,K-ATPase inhibitors in plasma of healthy individuals and in dialysis dependent patients. AB - We assayed plasma Na,K-ATPase inhibitory activity due to total lipids and lipid fractions. The effect of dialysis on the Na,K-ATPase inhibitory activity was also studied. Plasma lipid extracts from 11 healthy volunteers and 9 dialysis dependent patients (pre and post dialysis) were separated into neutral lipids and phospholipids. Further fractionation was by thin layer chromatography. These lipid fractions were analyzed for Na,K-ATPase inhibitory activity by displacement of [3H]-ouabain from hog brain Na,K-ATPase. Total inhibitory activity was significantly increased (p less than 0.001) in the post-dialysis plasma compared to pre-dialysis plasma of the same patient group and to controls (482, 85 and 78 nmol/L respectively; means of the groups in digoxin equivalents). The major inhibitory activity was associated with non-esterified fatty acids with modest contributions from four other lipid fractions. Our results show that endogenous lipids are major plasma Na,K-ATPase inhibitors in vitro under these assay conditions. PMID- 2550163 TI - Chemiluminescent detection of herpes simplex virus I DNA in blot and in-situ hybridization assays. PMID- 2550164 TI - 25-Hydroxyvitamin D and 1,25-dihydroxyvitamin D determined in serum by "SPE Octadecyl (C18)" column extraction and radioassay. PMID- 2550165 TI - Red cell membrane phosphatidylinositol kinase activity in hemolytic anemias and myeloproliferative diseases. AB - Phosphatidylinositol kinase activity was determined in red cell membranes from 85 healthy individuals, 20 patients with hereditary hemolytic anemia and 24 patients with myeloproliferative disorder. Increased activity was found in all ten cases of sickle red disease and seven among ten cases of other hereditary hemolytic anemias. These increases had no correlation with the reticulocyte count nor with the red cell shape. An unexpected decreased activity was found in several cases of myeloproliferative disorders, especially in polycythemia vera, with a negative correlation with the reticulocyte count. The mechanism(s) and significance of the phosphatidylinositol kinase abnormalities in these different groups of diseases remain to determine. PMID- 2550166 TI - Metabolic profiles in patients with insulinoma. AB - Twelve-hour metabolic profiles have been measured in six patients with insulinoma and results compared with normal subjects of similar age and weight. Fasting blood glucose was lower (mean +/- SEM 2.9 +/- 0.3 mmol/l vs 5.0 +/- 0.2 mmol/l) and plasma insulin higher (20.0 +/- 3.9 mU/l vs 7.2 +/- 1.6 mU/l) in insulinoma patients. Over the 12-h period blood glucose, pyruvate and glycerol were significantly lower, and plasma insulin, blood lactate, alanine and plasma non esterified fatty acids (NEFA) significantly higher in insulinoma patients. Overall the concentration of blood total ketone bodies was significantly higher in insulinoma patients. Values were higher in the early part of the day but lower later in the day and did not show the marked pre-meal rise observed in the normal subjects. The raised NEFA and ketone bodies are of particular interest as they may be a source of fuel supply in the presence of relative glucose deficiency. PMID- 2550167 TI - Beta-blockade disappearance rate predicts beta-adrenergic hypersensitivity. AB - We determined whether the beta-blockade disappearance rate would determine the degree of subsequent transient beta-adrenoceptor hyperresponsiveness after abrupt withdrawal of a beta-adrenoceptor drug. In a single-blind randomized study, 10 healthy men took a placebo for 1 week and then took nadolol one time a day (t1/2, 18 to 24 hours) or propranolol three times a day (t1/2, 4 to 6 hours) in doses that were increased weekly for 4 weeks to reach 240 mg per day. beta-Receptor responsiveness was assessed before and repeatedly after abrupt drug withdrawal by infusion of isoproterenol and epinephrine and by ergometer exercise. In the 13 days after drug discontinuation, peak beta-receptor sensitivity correlated (p less than 0.05) with the disappearance rate of beta-blockade as assessed by heart rate responses to isoproterenol (r = 0.68) and to submaximal exercise (r = 0.62) and by diastolic blood pressure responses to isoproterenol (r = 0.86) and epinephrine (r = 0.86). Plasma catecholamine levels and renin activity showed no overshoot. beta-Blockers with long plasma t1/2 values may prevent beta-blocker withdrawal syndromes by means of "self-tapering." PMID- 2550168 TI - Reversal of a calcium-mediated vasoconstrictor component in patients with congestive heart failure. AB - The influx of calcium into vascular smooth muscle cells is a major determinant of vasoconstriction, yet this concept has not been explored in congestive heart failure (CHF). We therefore used an "isolated" forearm model to assess the direct effects of the inhibition of calcium influx into vascular smooth muscle in 11 patients who had CHF, with use of the soluble dihydropyridine, nicardipine. Nicardipine produced a dose-dependent increase of forearm blood flow and a reduction of resistance, without producing a systemic hemodynamic effect. Patients with the lowest baseline forearm blood flow levels had the greatest percentage increases in forearm blood flow (r = -0.729, p less than 0.01), and a favorable metabolic effect was documented by a reduction in oxygen extraction across the forearm. This study demonstrated the importance of vascular smooth muscle intracellular calcium as a determinant of vasoconstriction in patients who have CHF. PMID- 2550169 TI - [Pathologic consequences of a severe influenza outbreak (swine virus A/H1N1) under natural conditions in the non-immune sow at the beginning of pregnancy]. AB - Pathological consequences of a severe outbreak of swine influenza (H1N1 virus) in the non immune sow at the beginning of pregnancy, under natural conditions. A sudden acute outbreak of fever, depression, anorexia and coughing in a group of nulliparous sows from a herd that was currently under epidemiological investigation lead to build a particular disposal of observation. The clinical signs were daily recorded including rectal temperature. Blood was taken from the sows at the beginning of the troubles and 3 weeks later for the detection of Aujesky's disease, coronavirus TGE-like, Influenza viruses A/H1N1 and A/H3N2 and Mycoplasma hyopneumoniae. Viral detection was attempted from nasal swabs and aborted fetuses during the acute phase. The clinical study showed fever reaching near 41 degrees C on most of the pigs and lasting usually from 2 to 5 days. The diagnosis of Influenza (virus swine H1N1) was established both on serology (massive seroconversion) and on the detection of the virus from the nasal swabs and from an aborted fetus. The control of the lungs of sows "not in pig" and culled showed extended lesions of bronchopneumonia and Pasteurella multocida was found. The technical consequences of this severe outbreak of Influenza on reproduction were mainly important at the beginning of pregnancy. Over 13 sows inseminated less than 1 week before the outbreak, only 3 farrowed (respectively 5.5 and 12 piglets); 7 returned to oestrus and 3 "not a pig" at 21 days (echotomography) did not show signs of heat and were culled. Over 8 pregnant sows (1 month of pregnancy), 6 farrowed normal litters and total embryonic resorption occurred in 2 sows. Over 18 pregnant sows (more than 45 days gestation) one aborted. PMID- 2550170 TI - Transfer of plasmid Hly in vivo in pigs intestine. AB - The results of our study suggest the in vivo transfer of Hly plasmid from native pathogenic and enterotoxigenic Escherichia coli strain to autochtonous Escherichia coli, using ileal loop test. To confirm this hypothesis pHly::Tn5 and PHly::Tn3 were obtained using an in vitro recombination method, and introduced to Escherichia coli laboratory strain. For experiments the laboratory strain, carrying pHly::Tn5 and pHly::Tn3 and pHly::Tn5 strain which acquired K88(F4) by means of conjugation, were used. In the study in which the donor Escherichia coli pHly::Tn5 strain, carrying antigen K88(F4), was injected into the ileum, pHly conjugants were isolated from faeces after 48 h in 2 out of 5 pigs, which was a low frequency. After the oral introduction of 10(9) cells of pHly::Tn5 and pHly::Tn3 Escherichia coli strains without the colonizing factor K88(F4), conjugants were not isolated from faeces of experimental animals. However when the pigs received donor CSH55 pHly::Tn5 Escherichia coli strain orally, which were also carrying plasmid K88(F4), transconjugants were obtained in a low frequency of 3 out of 9 pigs. Our experiments confirmed the suggestion of Smith that in vivo transfer of plasmid in the intestine of animals is only possible when the donor transfers the plasmid with high frequency and readily colonizes the intestine. The pHly::Tn5 plasmid acquired by in vitro recombination does not spread with time throughout the autochtonic population of Escherichia coli present in the intestine of swine. The results of our study showed the in vivo transfer in pigs intestine of Hly pathogenicity marker from both native pathogenic strains carrying antigen K88(F4) and constructed donor laboratory strain of Escherichia coli pHly::Tn5 also carrying antigen K88(F4) to autochtonous intestinal strains. PMID- 2550171 TI - Properties of the immunosuppressive factor induced by murine cytomegalovirus. AB - Murine cytomegalovirus infection of spleen cultures induced the production of a small (less than 10,000 molecular weight) immunosuppressive factor (VISF), which suppressed concanavalin-A mitogenesis in fresh mouse spleen cells, and in fresh human peripheral blood leukocytes. The factor did not affect the growth of two murine T-cell lines or of mouse fibroblasts. A similar factor was also found in the serum of infected mice, at the time of maximum immune suppression. The properties of VISF indicate that the mechanism of MCMV immune suppression is different from that caused by several other viruses which are important in human and veterinary medicine. PMID- 2550172 TI - Prevention of cancer: vegetables and plants. AB - 1. Results of epidemiological studies indicate that a human diet rich in vegetables may lower the incidence of cancer. 2. This preventive effect of the vegetable diet against cancer could be ascribed to lowered intake of energy (joules) and its content of vitamins and carotene. 3. The consumption of vegetables means also less meat and fats as well as increased fiber content and specific chemopreventive compounds (indoles, plant phenols) present in such a diet. 4. The supposed mechanisms of prevention may include enhanced enzymatic detoxification of harmful compounds, and inhibition of their binding to cellular DNA, their adsorption on fiber, detoxification of radical forms of carcinogens by natural antioxidants in plants and probably many other ways too. PMID- 2550173 TI - Type analysis of oligosaccharide chains on human and murine MHC class II alpha chains by the lectin-nitrocellulose sheet method. AB - 1. The microheterogeneous alpha molecules of class II antigen, DR molecules obtained from human B cell line and I-A molecules from mouse B cell hybridoma cell line, were separated by 2-D PAGE, transferred onto NC sheets and N-linked oligosaccharide types were analyzed by staining with P.O./lectins. 2. This is the first report to show directly the type of oligosaccharide chain corresponding to each spot separated by 2-D PAGE. The glycosylation patterns of class II alpha chains in human and mouse were compared. PMID- 2550174 TI - The prostaglandin synthesis in marine fish thrombocyte. AB - 1. Prostaglandin (PG) syntheses from labelled highly unsaturated fatty acids were investigated in washed thrombocyte suspensions of four species of marine fish, flounder (Paralichthys olivaceus), black seabream (Acanthopagrus schlegeli), black rockfish (Sebastes schlegeli), and red seabream (Pagrus major). 2. Synthesized PGs were analyzed by thin-layer radiochromatogram scanner and high performance liquid chromatography. 3. When [1-(14)C]arachidonic acid (AA) was incubated with washer thrombocyte suspension, AA was mainly converted to PGF(2alpha), PGE2, and PGD2 in all species. 4. It was suggested that [1 (14)C]eicosapentaenoic acid was mainly converted in PGs only in black rockfish thrombocytes, and the metabolites were mainly PGE3 and PGD3. 5 However, [1 (14)C]docosahexaenoic acid was not cyclized into PGs in all the species. PMID- 2550175 TI - Calpain and calpastatin levels in different organs of the rabbit. AB - 1. The levels of Ca-independent and Ca-dependent proteolytic activity as well as the activities of calpains and calpastatin in different organs of the rabbit was examined at various developmental stages. 2. Calpain and calpastatin levels were highest in the lung and in the kidney. 3. In all organs examined except the thymus the total level of calpain was higher than that of calpastatin. 4. In the thymus the levels of calpains and calpastatin decreased markedly with age. PMID- 2550176 TI - Multi-centre clinical trial of nedocromil sodium in reversible obstructive airways disease in adults: a general practitioner collaborative study. AB - The efficacy of nedocromil sodium (2 x 2 mg puffs twice a day) was assessed in a multi-centre, double-blind parallel group placebo-controlled study of 159 adult patients with chronic reversible obstructive airways disease. Over two-thirds of the patients had been maintained on or used an inhaled bronchodilator prior to the study. During a 4-week baseline period, patients stopped using their current respiratory therapy, with the exception of oral bronchodilators, and were established on a maintenance regimen of inhaled salbutamol (2 puffs 4-times daily with additional doses as needed). Patients then received either nedocromil sodium or placebo in place of the maintenance salbutamol regimen and inhaled salbutamol was allowed only on an as needed basis during a 12-week double-blind period. Daily diary records were made by patients of symptom severity, morning and evening peak expiratory flow rate, and salbutamol usage. The doctor assessed overall severity of the patient's condition at 4-weekly intervals, and both patients and doctor assessed treatment effectiveness at the end of the trial. Total symptom severity (night-time symptoms, morning tightness and daytime symptoms) was reduced and morning and evening PEFR improved throughout in the nedocromil sodium-treated patients, although evening PEFR returned nearly to the baseline in the final 4 weeks of the study. The reduction in combined day and night-time inhaled bronchodilator use from the baseline was significantly greater in the nedocromil sodium-treated patients and these patients were significantly (p less than 0.05) improved compared to the placebo-treated patients on each of the three occasions that the physicians assessed disease severity. Of the 40 patients withdrawn during the double-blind period of the study (26 on nedocromil sodium, 14 on placebo), 9 and 7 patients, respectively, were withdrawn because of the onset of uncontrolled asthma, and 4 and 2 patients, respectively, because of suspected adverse reactions to treatment. A total of 21 patients treated with nedocromil sodium recorded unusual symptoms (9 commented on taste, 5 had nausea and vomiting) compared with 16 patients on placebo. Overall, nedocromil sodium was considered to be moderately or very effective in at least 50% of the patients, although no significant differences were seen between treatments in either the patients' or physicians' assessment. PMID- 2550177 TI - Assignment of the human myeloperoxidase gene (MPO) to bands q21.3----q23 of chromosome 17. AB - Using a human myeloperoxidase cDNA, we have mapped the human myeloperoxidase gene to chromosome 17 at q21.3----q23 by in situ hybridization to metaphase chromosomes from human lymphocyte preparations. PMID- 2550178 TI - DNA base sequence is not the only factor for restriction endonuclease activity on metaphase chromosomes: evidence using isoschizomers. AB - Human and mouse fixed metaphase chromosomes were treated with the isoschizomer sets MboI/Sau3A and EcoRII/BstNI. In both cases we found that each member of the isoschizomer pairs produced different results, indicating that factors other than DNA base composition may affect in situ digestion by restriction endonucleases and that the structure of the enzymes is one factor. We also found that MboI and Sau3A isoschizomers produced the same effect on the chromosomes of the grasshopper Oedipoda germanica. This indicated that differences in the chromatin structure of different species may be important in determining restriction endonuclease activity on eukaryotic chromosomes. PMID- 2550179 TI - Mitotic deletions of 11p15.5 in two different tumors indicate that the CALCA locus is distal to the PTH locus. AB - We have compared the constitutional and tumor genotypes in two patients with Wilms tumor and adrenocortical carcinoma. The allelic distribution of chromosome 11-specific markers spanning chromosome 11 from pter to qter (HRAS1-HBB [CALCA/PTH]-FSHB-CAT-APOA1) and an approach combining RFLP analysis and gene copy number determination showed that a mitotic deletion had occurred in both tumors. The loss of one copy of the gene for alpha-calcitonin-gene-related polypeptide (CALCA), together with that of a more distal marker (HRAS1 or HBB), indicates that CALCA is distal to the gene for parathyroid hormone (PTH), which was not deleted in either tumor. These results suggest that mitotic deletion mapping may be as useful as meiotic deletion or recombination mapping in ordering closely linked markers, such as CALCA and PTH, for which other approaches, including physical mapping and multipoint linkage analysis, have failed to accurately identify the gene order. PMID- 2550180 TI - Analysis of complex Y chromosome aberrations using a single DNA probe (Y-367). AB - A specific cloned DNA sequence (Y-367) detects at least four loci in the euchromatic long arm and in the short arm of the human Y chromosome. Deletion mapping assigns one locus to the distal euchromatic long arm, another to a region close to the centromere on either Yq or Yp, and two additional loci to the Y short arm. Y-367 may thus be used for the rapid screening of even complex Y chromosome aberrations. This is exemplified in a 45,X male with Y chromosome material on the long arm of chromosome 10 by the detection of an inversion of a portion of Yp and by the confirmation of duplications and deletions in two individuals with duplications of part of the Y chromosome. PMID- 2550181 TI - Neuroectodermal tumor versus Ewing's sarcoma--immunohistochemical and electron microscopic observations. PMID- 2550182 TI - The cytogenesis of macrophages and osteoclast-like giant cells in bone tumors with special emphasis on the so-called fibrohistiocytic tumors. AB - Recent investigations have suggested that osteoclasts and osteoblasts belong to different cell systems: osteoclasts originate from hemopoietic stem cells, most probably via precursors of the mononuclear phagocyte system. Osteoblasts, however, arise from local mesenchyme. The present classification of bone tumors issued by the WHO, however, is still based on the assumption of osteoclasts and osteoblasts being merely different manifestations or differentiations of the same basic cell type. Consequently, histiocytes or macrophages as well as osteoclast like giant cells are interpreted in most bone tumors as an autochthonous component of the tumor. In the present study, this theory is contradicted by histological immunohistological, electron microscopic, and autoradiographic electron microscopic results on a larger number of osteosarcomas, chondromas, chondrosarcomas, chondroblastomas, aneurysmal bone cysts, giant cell tumors of bone, malignant fibrous histiocytomas, fibrosarcomas, desmoplastic fibromas, Ewing's sarcomas, fibrous dysplasias, nonossifying fibromas, and malignant hemangioenkdotheliomas of bone. In order to elucidate the role of macrophages and osteoclast-like giant cells, different monoclonal antibodies were applied to bone tumor specimens as markers of mononuclear macrophages and giant cells. The concept of what is called fibrohistiocytic tumors should be reconsidered. Immunohistological studies have shown that in malignant fibrous histiocytoma as well as in giant cell tumors of bone only a certain portion of macrophages will react with the highly specific antibodies, whereas the majority of tumor cells are negative. This finding alone suggests that the infiltration of macrophages is a reactive phenomenon. It is in agreement with earlier autoradiographic and electron microscopic investigations on giant cell tumors of bone, confirming that only the fibroblast-like tumors are actually proliferating. These data were verified in the present study with the aid of double labeling immunohistological techniques, using antibodies against mature tissue macrophages and others against a proliferation-associated nuclear antigen. Only the fibroblast-like cells, which do not react with the macrophage-specific antibody, will express the proliferation-associated nuclear antigen. Analogous results were obtained in malignant fibrous histiocytoma. We may infer from these results that the majority of these tumors must be neoplasms of local mesenchyme, mostly in fibroblastic differentiation, while the considerable number of macrophages is seen as a reactive phenomenon.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2550183 TI - Acquired cytomegalovirus infection of breast milk in infancy. AB - A cross-sectional study was conducted to examine the rate of cytomegalovirus (CMV) shedding through breast feeding and the rate of acquired infection by the infants. Among the 104 seropositive breast-feeding mothers, CMV was recovered in the milk in 19 cases, with a milk shedding rate of 18.3%. Isolation of CMV from the urine of the infants of the 188 seropositive mothers found 36 positive cases. Of the 36 cases, 15 of 19 fed on CMV-positive milk were infected through breast feeding, with an infection rate of 78.9%; 11 of the 85 fed on CMV-negative milk were infected, the infection rate being 12.9%; and 10 (11.9%) of 84 fed on cow's milk were infected. These demonstrate that the rate of infection in infants fed on virus-positive milk was significantly higher than those fed on virus-negative milk or cow's milk (P less than 0.005). PMID- 2550184 TI - Viral etiology of cervical carcinoma. Human papilloma virus and herpes simplex virus type 2. AB - The possible role of human papilloma virus (HPV) and herpes simplex virus type 2 (HSV-2) in the viral etiology of cervical carcinoma was investigated a series of cervical lesions were studied for the presence of HPV and HSV-2 DNA sequences as well as HPV and HSV-2 antigens by DNA dot blot hybridization technique and high specificity PAP staining method. The results obtained were correlated with the histologic diagnosis. HPV 16 DNA sequences detected in cervical carcinoma biopsies were 43%, whereas HSV-2 DNA sequences were only 8%. HPV antigens detected in cervical dysplasia were 31%, whereas those detected in cervical carcinoma and cervicitis were the least. HSV-2 antigens were detected in chronic cervicitis, dysplasia and cervical carcinoma. The difference in positive rate between the cervical carcinoma and cervicitis groups was statistically significant, (chi-square test, P less than 0.01). No HPV DNA and HSV-2 DNA sequences were found in the same specimen, although both HPV DNA sequences and HSV-2 antigens were found in the same sample in some cases. The results indicate that the viral etiology of cervical carcinoma may be multifactorial. Both HSV-2 and HPV may be associated with cervical carcinoma, but the mechanisms involved are different. HSV-2 and HPV may act synergistically in the development of cervical carcinoma. PMID- 2550185 TI - [Inhibition by VIP of the proliferation of pancreatic cancer cells]. AB - The molecular biological mechanism of the inhibition by VIP (vasoactive intestinal peptide) on proliferation of pancreatic cancer cells was studied by identifying hormone receptor and using tumor model in vivo. Inhibition appears only in those cells with VIP receptor. Cell-membrane receptors may change during carcinogenesis process. The inhibition of VIP (at certain concentration) on proliferation of tumor cells is remarkable, but, on the other hand, negligible on that of normal cells. These results suggested the feasibility of clinical application of VIP to the treatment of human pancreatic cancer. PMID- 2550186 TI - [Resection of hepatic hilar liver cancer]. AB - From January 1970 to January 1987, hepatic hilar liver cancer (or central type of hepatic cancer) resection was done in 51 cases. The resection was often extremely difficult, and sometimes complete occlusion of the hepatic blood supply was needed when the tumor lied close to, or already invaded the great blood vessels. The 1-, 3-, and 5- year postoperative survival rates were 65.7%, 45.3%, and 38.8%, respectively, in contrast to 93.8%, 86.1%, and 80.1% (P less than 0.001) obtained in 51 cases of peripheral type liver cancer of similar tumor size during the same period. It is considered that the difficulty involved in the resection of such a cancer, the limited extent of resection, and the easy entrance of tumor cells into the blood stream may explain the poor prognosis. PMID- 2550187 TI - Urinary excretion of angiotensin-converting enzyme in NIDDM patients with nephropathy. AB - Twenty-four-hour urinary excretion of angiotensin-converting enzyme (ACE) was investigated in relation to that of albumin and beta 2-microglobulin (beta 2M) in 25 non-insulin-dependent diabetes mellitus (NIDDM) patients without nephropathy, 13 NIDDM patients with incipient nephropathy, 18 NIDDM patients with overt nephropathy, and 14 nondiabetic subjects. NIDDM patients without nephropathy and nondiabetic subjects were similar in albumin, beta 2M, and ACE excretion. NIDDM patients with incipient nephropathy had elevated albumin excretion (P less than .01) and similar beta 2M and ACE excretion compared with nondiabetic subjects. On the other hand, NIDDM patients with overt nephropathy had elevated albumin, beta 2M, and ACE excretion compared with nondiabetic subjects (P less than .01). In all NIDDM patients studied, a positive correlation was found between ACE excretion and albumin excretion (r = 0.76, P less than .001) or beta 2M excretion (r = 0.52, P less than .01). These data suggest that elevated ACE excretion in NIDDM patients with overt nephropathy may be reflective of renal tubular damage. PMID- 2550188 TI - Mucinous breast carcinoma and mixed mucinous-infiltrating ductal carcinoma: a comparative cytologic study. AB - In its pure form, mucinous breast carcinoma (MC) has a much better prognosis than infiltrating ductal carcinoma (IDC). Mixed MC-IDC has the prognosis of IDC. We compared the fine-needle aspiration (FNA) cytology and histology of nine cases of pure MC with 13 cases of mixed MC-IDC. While typical of pure MC, abundant mucin (3+/3+) was noted in smears from three cases of mixed tumor and is thus necessary but not sufficient for a diagnosis of pure MC. Cellular pleomorphism has been said not to be a feature of MC; however, we found occasional large cells markedly different from the typical small uniform cells of MC in four pure and seven mixed tumors. Cytologic features indicative of a mixed tumor include one or more smears totally without mucin, scantly amounts of mucin, or necrosis. A combination of features indicative of pure mucinous carcinoma in FNA included abundant mucin on all smears, no pleomorphism, and no necrosis. Many cases will have smear patterns that are not typical of either profile and should probably be designated as carcinoma with a mucinous component. PMID- 2550189 TI - Glial fibrillary acid protein immunoreactivity in fine-needle aspiration of salivary gland lesions: a useful adjunct for the differential diagnosis of salivary gland neoplasms. AB - The value of immunocytochemical staining for glial fibrillary acid protein (GFAP) in salivary gland lesions was investigated in 33 fine-needle aspiration smears. The study utilized cytologic material from ten pleomorphic adenomas, six normal salivary glands, three cases of chronic sialadenitis, three Warthin's tumors, two adenoid cystic carcinomas, three adenocarcinomas, two malignant mixed tumors, one acinic cell carcinoma, and three mucoepidermoid carcinomas. All tested pleomorphic adenomas stained positively. The adenoid cystic carcinomas and the cases of chronic sialadenitis, along with the low-grade mucoepidermoid carcinoma, were negative for GFAP immunoreactivity. These results indicate that immunostaining for GFAP may be a valuable aid in the diagnosis of pleomorphic adenoma; GFAP may be especially helpful in distinguishing those cases for which the differential diagnosis includes the aforementioned salivary gland neoplasms. PMID- 2550190 TI - Mucinous metaplasia to neoplastic lesions in endometrial samples with cytohistologic correlation. AB - Seven cases of endometrial mucinous metaplasia and five of well-differentiated mucinous adenocarcinoma of the endometrium were studied. Cytologic specimens were obtained by Isaacs endometrial sampler, avoiding endocervical contamination. The histologic diagnosis between the intermediate type of mucinous metaplasia and the well-differentiated mucinous adenocarcinoma posed no problems. DNA analyses of the histologic samples showed a euploid pattern in benign and intermediate types of metaplasia, while well-differentiated mucinous adenocarcinoma showed a hyperdiploid pattern. The cytologic diagnosis of benign mucinous metaplasia should be suggested in the presence of abundant mucinous cells in endometrial samples in the absence of nuclear abnormalities. PMID- 2550191 TI - Fine-needle aspiration biopsy of clear-cell sarcoma of the kidney: light and electron microscopic features. AB - Smears from seven fine-needle aspiration biopsies from three patients with clear cell sarcoma of the kidney were reviewed, and the findings were correlated with corresponding histopathologic appearances. In two cases, the tumor cells were polygonal to spindle-shaped and were loosely arranged in a matrix of mucoid material. The histopathologic examination of the resected tumors in these cases revealed a classic pattern. In the third case, the aspiration smears revealed round to polygonal cells with moderate to abundant cytoplasm but without the mucoid matrix. The predominant histologic appearance of this tumor was an epithelioid trabecular pattern. Electron microscopic findings were similar to those described in published studies. PMID- 2550192 TI - Chain of events in Papanicolaou smear testing: impact on quality assurance. AB - Quality assurance is defined in this article as a comprehensive program aimed at preventing and controlling errors from the time a test is ordered until test results are reported and utilized in the care of the patient. Quality control is only one facet of a total quality assurance program. Recently, the general public, professional organizations, and regulatory agencies have scrutinized the standards and practice of Papanicolaou smear testing. The components of quality assurance for gynecologic cytology are reviewed, and the chain of events in performing effective cytologic testing is described. The impact of various events on the outcome of Papanicolaou smear testing is thoroughly discussed. PMID- 2550193 TI - Recombinant human erythropoietin produced by Namalwa cells. AB - To establish a practical exogenous gene expression system in human cells, a cDNA coding for human erythropoietin (EPO) was expressed in human B-lymphoblastoid Namalwa cells. The Namalwa-derived recombinant EPO was purified from the culture fluid by a simple three-step procedure. The Namalwa EPO showed an equivalent activity in vivo to that of human urinary EPO. Oligosaccharide structure analyses suggested that almost all N-linked oligosaccharide chains of Namalwa EPO are shared by urinary EPO. The two major N-linked oligosaccharides of Namalwa EPO were fucose-containing tetraantennary and fucose-containing triantennary structures. PMID- 2550195 TI - Adherence to high-carbohydrate, high-fiber diets. AB - Successful diabetes management demands a high degree of adherence. Adherence to medical prescriptions, particularly special diets, is notoriously poor. A study of 40 individuals with IDDM and NIDDM who were followed an average of 23 months found good or excellent dietary adherence in 70% of subjects; only 5% demonstrated poor adherence. Adherence was assessed objectively based on carbohydrate, fat, and fiber intakes reported in 24-hour food recalls, 7-day food frequency surveys, and home food records. High dietary fiber intake may independently enhance adherence, perhaps by increasing satiety or because of the simplicity of the concept. Good communication between the health professional and patient is also fundamental to all stages of dietary adherence. Use of adherence enhancing techniques throughout adoption of new diet behaviors promotes long-term adherence to high-carbohydrate, high-fiber diets. PMID- 2550194 TI - Replica filter screening technique to detect transfected cells expressing beta 2 adrenergic receptor. AB - We have utilized a replica transfer technique to develop a novel screening assay for the identification of transfectants expressing beta 2-adrenergic receptors (beta 2-AR). The hamster beta 2-AR gene flanked by either its natural promoter or the zinc-inducible mouse metallothionein (MMT) promoter was cotransfected with plasmids conferring neomycin resistance (pRSVneo) into beta 2-AR-deficient mouse L cells. Transfectant colonies were grown on polyester nylon filters and screened by filter binding with [125I]iodohydroxybenzylpindolol to identify colonies expressing beta 2-AR. Individual colonies were isolated and examined to determine beta 2-AR gene dosage, mRNA expression, and receptor densities and affinities. Analysis of cell lines expressing beta 2-AR indicates that this method can identify transfectants containing only a single beta 2-AR gene copy and expressing as few as 4,000 beta 2-receptors per cell. This method may be useful as a tool for the molecular cloning of neurotransmitter receptor genes and for the measurement of transfection efficiencies and expression of receptor genes in cells. PMID- 2550196 TI - [Spontaneous metastasis of clonal cell subpopulations of human lung giant cell carcinoma after subcutaneous inoculation in nude mice]. AB - Four sub-lines (strain A, C, D, E) were isolated from a human lung giant cell carcinoma cell line (PLA-801) by the single cell cloning technique. The incidence of spontaneous metastasis was higher in strain D, moderate in strains A, E and lower in strain C after they were inoculated subcutaneously into the nude mice. The lung and lymph-nodes were the major target organs of metastasis. Morphological studies (light microscope, electron microscope and immunohistochemistry) showed that the cytoplasm of metastatic tumor cells was rich in microfilament and positive for Vimentin. The results strongly support the theory on tumor cell heterogenicity. In addition, these clones, especially strains D and C, could be an ideal model for the study of tumor metastasis. PMID- 2550197 TI - [A newly recognized precancerous lesion of the stomach--histopathologic features of globoid dysplasia of human gastric epithelium]. AB - Histopathologic features of globoid dysplasia of human gastric epithelium were described by means of observation of serial paraffin sections of 53 cases of globoid dysplasia. It was divided into three grades according to the architecture and cellular atypia. Penetration of outer layer globoid dysplastic cells through the basement membrane of "double layers structure" appeared in typical globoid dysplasia Grade III and infiltration of globoid dysplastic cells into stroma as well as the formation of incipient focus of signet ring cell carcinoma were described. The twinkling scene of infiltration of the globoid dysplastic cells into lamina propria through the basement membrane and the damage of basement membrane by globoid dysplastic cells were shown by Gordon Sweet's stain. Through the analysis of background lesions of the globoid dysplasia, a conclusion can be made that the globoid dysplasia might be an important precancerous lesion of the signet ring cell carcinoma of the stomach. PMID- 2550198 TI - [Clinico-pathologic comparison between early gastric cancer and early cancriform progressive cancer]. AB - Clinico-pathologic comparison between 36 cases of early gastric cancer (EGC) and 34 cases of early cancriform progressive cancer (ECPC) were made. It was found that there was no difference in the average age and sex. Epigastric pain occurred in most of the cases. Weight loss, hematemesis and melena occurred more frequently in patients with ECPC than those with EGC. Pathologically, the cancers were located in gastric antrum in most of the cases in both groups. The lesions 1 2 cm across accounted for half in each. Lesions over 4 cm (19.4%) was obviously more common in EGC than in ECPC (2.9%). Macroscopic classification showed that though pouch-type predominated in both groups, it was more frequent in ECPC (88.2%) than in EGC (72.2%). Histologically, there were more highly differentiated types in EGC (52.7%) and more poorly differentiated types in ECPC (52.9%). The Pm depth of ECPC was high. The prognosis of ECPC was favorable, second only to EGC. ECPC belongs to Type 5. PMID- 2550199 TI - [Radical total gastrectomy and improved operation in gastric cancer]. AB - 1089 patients with gastric cancer resected from 1979-1986 are analysed. Of 1089 patients, 105 underwent total gastrectomy, including 55 by routine method as control and 50 by improved method as study group. The improved total gastrectomy was to take the jejunum instead of a gastric pouch for reconstruction of alimentary tract. End-to-side anastomosis was carried out by stapling devise, forming a reversed "7" gastric substitute and ensuring blood supply as well as physiologic patency and less postoperative complications. This technique is simple and easy to be adopted. The incidence of complication was 9% without operative mortality in this series. The 5-year survival rate was 35.75%. PMID- 2550201 TI - [Results of bronchial artery chemotherapy infusion in lung cancer--report of 82 cases]. AB - Eighty-two patients with lung cancer confirmed by cytology and/or histology were treated by bronchial artery chemotherapy infusion. There were 61 males and 21 females. The ages ranged from 30 to 75 years with an average of 54.6. Histologic types were 48 squamous cell carcinomas (58.5%), 20 adenocarcinomas (24.4%), and 14 undifferentiated small cell carcinomas (17.1%). TNM classification showed 28 Stage IIIa, 32 Stage IIIb and 22 Stage IV. Sixty-nine patients were treated by Cis-platinum combined with cyclophosphamide (84.1%). The results showed that 20 patients had complete response (24.4%), 28 partial response (34.1%), 25 stability (30.5%), 8 progression (9.8%) and 1 died (1.2%). Ten patients underwent lung resection after infusion of drugs. The factors influencing prognosis and complications are discussed. PMID- 2550200 TI - [Post-operative pleural infection of lung cancer and its influence on survival]. AB - 1139 patients with primary lung cancers resected from 1954 to 1984 are reported. Twelve of them had pleural infection post-operatively, an infection rate of 1%. The infection caused mainly by Staphylococcus aureus developed on Days 5-23 post operatively. Most of the patients had fever. Closed drainage was applied for each case. Two of these 12 patients survived for more than 5 years. They had Stage 1 and Stage 2 small cell carcinoma and adenocarcinoma. No significant difference was observed in the 5-year survival rate (33.3% and 30.6%) between the infected group and uninfected group. There was no relation between the post-operative pleural infection and prognosis. With review of related literature, beneficial immunologic response leading to better prognosis might exist in the infected cases, but further observation and discussions are needed. PMID- 2550202 TI - [Ceruminal gland tumor--report of 7 cases and review of literature]. AB - Seven patients with ceruminal gland tumors originating from the external auditory canal are reported. The review of literature of this tumor showed that ceruminal gland tumors are commonly considered to arise from ceruminal gland but also from heterotopia salivary gland in the external auditory canal as believed by others. The term "ceruminal adenoma or adenocarcinoma" is still used by many authors. Histologically, this tumor is divided into four subtypes: adenoma, pleomorphic adenoma, adenocarcinoma and adenoid cystic carcinoma. Different treatments should be given for this tumor according to their different benign or malignant degrees. PMID- 2550203 TI - [Innervation of beta-adrenoreceptors of human skin blood vessels]. PMID- 2550204 TI - Antagonism of aminoglutethimide and adrenocorticotropic hormone (ACTH) studied on slices of adrenal glands of the guinea-pig. AB - Aminoglutethimide at concentrations from 0.1 to 5 nM is able to inhibit the cortisol release elicited by adrenocorticotropic hormone (ACTH) (from 2.5 to 50 ng/ml) in guinea-pig adrenal cortex slices. The antagonism is a non-competitive one (in a Lineweaver-Burk plot), whereas other drugs (morphine, endorphin, indomethacin, etc.) inhibit ACTH competitively. This is in agreement with the known mechanism of action of aminoglutethimide, which inhibits the synthesis of cortisol by blocking reactions of enzymes such as aromatase and desmolase. From the data one can calculate the dissociation constant (Km) of ACTH with its receptor(s) to be 0.27 pg/ml and the inhibiting constant (Kl) of aminoglutethimide to be 49.78 x 10(-10) M. The maximal response of ACTH was 52.9 ng/ml. PMID- 2550205 TI - [Lung function following irradiation in pediatric cancer patients]. AB - Lung function tests were performed in 22 children and juveniles who had received radiotherapy to the lungs, an average of 9.5 years previously, for tumour (14 with Hodgkin's disease [aged 7-22 years], 4 with malignant non-Hodgkin lymphoma [aged 6-14 years], 3 with Wilms tumour [4-6 years], and one with Ewing's sarcoma [aged 16 years]). All three patients who, as young children, had had radiotherapy to both lungs because of a Wilms tumour with multiple lung metastases had restrictive disorders of lung function. Four of 12 after treatment of Hodgkin's disease and one of two after malignant non-Hodgkin lymphoma and extensive thoracic irradiation developed a restrictive disorder of pulmonary function. But all four patients who had irradiation restricted to the mediastinum had normal lung functions. PMID- 2550207 TI - Growth hormone releasing hormone-sensitive adenylate cyclase activity in growth hormone-producing pituitary adenoma: correlation to the response of plasma growth hormone to growth hormone releasing hormone in patients with acromegaly. AB - The correlation between response of plasma GH to GHRH and the GHRH-induced stimulation of the intracellular adenylate cyclase (AC) activity in pituitary adenoma cell membranes in acromegalic patients was investigated. Each peak plasma GH level after iv administration of GHRH ranged from 1.1 to 13.8 times the basal level in 13 acromegalic patients. On the other hand, the maximal stimulation of intracellular AC activity (cAMP production) induced by GHRH varied from 1.4 to 6.4 times the control level in each GH-producing pituitary adenoma cell membrane. A significant positive correlation (r = 0.89, P less than 0.005) between plasma GH response to GHRH and intracellular cAMP production stimulated by GHRH was observed in nine of the acromegalic patients. In contrast, the response of plasma GH to GHRH was significantly blunted, despite a fairly large production of intracellular cAMP stimulated by GHRH, in the other four acromegalic patients. These results suggest that GHRH-induced GH release from GH-producing pituitary adenomas of patients with acromegaly may be regulated not only by GHRH receptor adenylate cyclase system but also modified by several other factors including somatostatin and Sm-C. PMID- 2550206 TI - [The occurrence of a reovirus variant in a German broiler flock]. AB - Broilers deriving from a parent flock, which had been effected in the 6th. month of hatching egg production, show arthritis beginning with the 12th day of life. The tarso-metatarsal joint has been affected. Birds show stunting. Body weights at slaughter and feed conversion of the affected flocks were reduced. The percentage of condemned birds before slaughter was highly increase and came up to 3-5%. Chickens of other breeder flocks, which were reared with the diseased birds, showed viral arthritis at an age of 18-20th day of life. The boilers derived from parent flocks which had been vaccinated twice during the with a 1133 reo live vaccine and before laying with an oil based vaccine of the antigen type WVU. A reovirus has been isolated (isolate K 171/87), which caused viral arthritis in 1133-immune day old chicks after parenteral and oral application. Infection of these chickens with the pathogenic reovirus of the antigen type 1133 didn't cause a disease. Also by serological examinations it was shown, that the reo-isolate K 171/87 possesses a different antigenicity. The kind of occurrence indicates, that this reovirus infection has been transmitted vertically from one parent flock and it spread laterally to chickens of other parent flocks in broiler farms. PMID- 2550208 TI - ACTH determination in a petrosal sinus venous specimen after corticotrophin releasing factor provides the best clue on the laterality of microadenoma in Cushing's disease. AB - A 42-year-old woman with Cushing's disease was endocrinologically cured after transsphenoidal selective excision of a left-sided microadenoma. Simultaneous samplings from the bilateral inferior petrosal sinuses with ovine corticotrophin releasing factor (CRF) stimulation were performed preoperatively to evaluate some relations between the localization of a possible microadenoma and the levels of ACTH in inferior petrosal sinuses. The data for the venous samplings were as follows:-(1) The basal levels of ACTH obtained simultaneously from both inferior petrosal sinuses and peripheral vein were about the same. (2) A significant difference, with a high level of ACTH from the ipsilateral side of the microadenoma, was demonstrated after CRF stimulation. (3) The magnitude of increase in ACTH on the contralateral side after CRF was similar to that of the peripheral response. It is recommended that all patients with ACTH dependent Cushing's syndrome and negative radiological findings should have bilateral simultaneous inferior petrosal venous sampling with CRF stimulation. PMID- 2550209 TI - Serum parathyroid hormone concentration measured by highly sensitive assay in post-thyroidectomy hypocalcemia of patients with Graves' disease. AB - To investigate the role of parathyroid function in transient hypocalcemia after subtotal thyroidectomy for Graves' disease, the serum parathyroid hormone (PTH) concentration and nephrogenous (N) cAMP were measured in 16 patients before and after surgery. Serum PTH was measured with two commercially available kits (PTH M, PTH-C), PTH-M is a recently developed highly sensitive assay using an antibody recognizing the mid-portion of human PTH and a synthetic 125I-tyr45-human PTH (43 68) as a radioligand. One of the 16 patients had severe clinical tetany and had a markedly lower PTH-M concentration and NcAMP after thyroidectomy. However, no significant change in serum PTH-M, PTH-C and NcAMP were observed in the other patients, although their serum calcium (Ca) concentrations decreased significantly. The Data were analyzed by dividing the patients according to the change in serum Ca or PTH. Serum PTH-M and PTH-C significantly decreased in 4 patients whose serum Ca clearly decreased after surgery. Serum Ca on the first postoperative day was significantly lower in patients whose serum PTH decreased after thyroidectomy than in patients whose serum PTH did not. Furthermore, the serum Ca concentration was significantly correlated with PTH-M, and with NcAMP on the third postoperative day. These data proved that hypofunction of the parathyroid gland is important in transient hypocalcemia after subtotal thyroidectomy for Graves' disease. The pathogenetic mechanism of transient hypocalcemia was discussed in comparison with the data from a patient who had overt parathyroid injury. PMID- 2550210 TI - A new variant of 17 alpha-hydroxylase deficiency with hyperaldosteronism in two Japanese sisters. AB - We present a report on two sisters who have 17 alpha-hydroxylase deficiency with hyperaldosteronism. They have hypertension and hypergonadotropic hypogonadism. The steroid profiles suggest that they have 17 alpha-hydroxylase deficiency. In contrast to the classical biochemical findings in 17 alpha-hydroxylase deficiency, both of these patients have hyperaldosteronism. Thus this report describes a new variant of 17 alpha-hydroxylase deficiency with hyperaldosteronism. Dexamethasone suppressed the mineralocorticoid excess, including aldosterone, and improved their hypertension. In the untreated state, ACTH, instead of the renin-angiotensin system, regulated plasma aldosterone levels, but during dexamethasone treatment the renin-angiotensin system regulated these levels. PMID- 2550211 TI - Bronchoscopic "resection" of small cell carcinoma. AB - Small cell carcinoma of the bronchus frequently presents as a widely disseminated tumor with a large central primary lesion. A patient with peripheral shadowing on a chest radiograph is reported in whom bronchoscopy revealed endobronchial tumor of the small cell type. The patient underwent lobectomy, but residual carcinoma could not be identified in the operative specimen, and the patient has remained disease-free for 2.5 years post-resection. PMID- 2550212 TI - Solitary Peutz-Jeghers type polyp of the stomach in the absence of familial polyposis coli in a teenage boy. AB - We treated a male teenager with a solitary gastric Peutz-Jeghers type (hamartomatous) polyp showing neither mucocutaneous pigmentation nor inheritance. X-ray studies of the upper gastro-intestinal tract, small intestine and barium enema revealed a solitary thumb tip-sized polypoid mass in the posterior wall of the gastric body, with no evidence of any associated lesion. Histologically, the polypoid mass showed typical features of a Peutz-Jeghers type (hamartomatous) polyp. This may be the first documentation of a "solitary" gastric Peutz-Jeghers type polyp, in the absence of familial polyposis coli and mucocutaneous pigmentation. PMID- 2550213 TI - Retinol-binding protein: the serum transport protein for vitamin A. AB - The information available regarding the chemical structure of RBP, the structure of the RBP gene, and the expression of the RBP gene has expanded dramatically in recent years. Still many questions concerning RBP remain to be answered. The longstanding and important questions concerning the possible existence and the biochemical characteristics of the RBP cell surface receptor are in need of resolution. The factors that regulate RBP secretion from the liver still remain to be fully elucidated. Additional information concerning the physiological role of RBP synthesis in extrahepatic tissues is needed. Considering what is now known, it is clear that in the future much intense research will be required before the many important questions regarding the structure, synthesis, secretion, and physiological roles of RBP can be answered. PMID- 2550215 TI - The involvement of the endocrine system in regulating cardiovascular function: emphasis on vitamin D3. PMID- 2550214 TI - The G-protein family and their interaction with receptors. PMID- 2550216 TI - Antiepileptic drug actions. AB - Antiepileptic drugs (AEDs) vary in their efficacy against generalized tonic clonic, myoclonic, and absence seizures, suggesting different mechanisms of action. Phenytoin (PHT), carbamazepine (CBZ), and valproate (VPA) reduced the ability of mouse central neurons to sustain high-frequency repetitive firing of action potentials (SRF) at therapeutic free serum concentrations. Phenobarbital (PB) and the benzodiazepines (BZDs), diazepam (DZP), clonazepam (CZP), and lorazepam (LZP), also reduced SRF, but only at supratherapeutic free serum concentrations achieved in treatment of generalized tonic-clonic status epilepticus. These AEDs interact with sodium channels to slow the rate of recovery of the channels from inactivation. The BZDs and PB enhanced gamma aminobutyric acid (GABA) responses evoked on mouse central neurons by binding to two different sites on the GABAA receptor channel. BZDs increased the frequency of GABA receptor channel openings. In contrast, barbiturates increased the open duration of these channels. VPA enhanced brain GABA concentration and may enhance release of GABA from nerve terminals. Ethosuximide (ESM) reduced a small transient calcium current which has been shown to be involved in slow rhythmic firing of certain neurons. Reduction of SRF, enhancement of GABA-ergic inhibition, and reduction of calcium current may be, in part, the bases for AED action against generalized tonic-clonic, myoclonic, and absence seizures, respectively. PMID- 2550217 TI - Effect of uridine on hepatic galactose-1-phosphate uridyltransferase. AB - Uridine sugar nucleotides are important intermediates in galactose metabolism and may play a role in the long-term galactose toxicity in human galactose-1 phosphate uridyltransferase deficiency galactosemia. Since administration of uridine, a precursor of uridine nucleotides, has been considered as a therapeutic measure, we have investigated the effects of this compound on the activity of rat hepatic transferase. Uridine has been found to be an inhibitor of the enzyme in in vitro studies and to cause an increase in galactose-1-phosphate in liver perfused with galactose which is consistent with physiologic inhibition of the enzyme. Uridine is a partial linear competitive inhibitor of UDPglucose and an uncompetitive inhibitor of galactose-1-phosphate. These findings suggest caution should be applied in giving the compound to subjects with genetically limited transferase activity because of the possibility of inhibiting the small amount of residual enzyme. PMID- 2550218 TI - Isolation of A/Equi-2 virus during 1987 equine influenza epidemic in India. AB - Processing of nasal materials from clinical cases during the 1987 influenza epidemic in Northern and Central India resulted in the isolation of two haemagglutinating agents; one each from donkeys and horses at Bhiwani in Haryana State and Ludhiana in Punjab State, respectively. These were typed as Influenza A/Equi-2 viruses by haemagglutination inhibition test. The two isolates were designated as A/Equi-2/Bhiwani/1/87 and A/Equi-2/Ludhiana/1/87. The Bhiwani/87 isolate was confirmed to have H3N8 antigenic structure and was indistinguishable from the Miami/63 strain of A/Equi-2 virus. However, the A/Equi-2 Ludhiana/87 isolate was closely related to the Fontainebleau/79 strain of A/equi-2 virus. PMID- 2550219 TI - Laboratory diagnosis of primary hyperparathyroidism. AB - Although considerable effort has gone into the development and modernization of indirect parathyroid function tests, recent experience indicates that they do not possess adequate diagnostic specificity or sensitivity to provide a definitive diagnosis of HPT when they are most needed, that is, in mild or intermittent hypercalcemia. Conversely, in most research and some commercial laboratories, the combination of hypercalcemia and an elevated mid-molecule or intact PTH level will be diagnostic in more than 90% of patients. PMID- 2550220 TI - Identification and properties of a novel type of Na+-permeable amiloride sensitive channel in thyroid cells. AB - Amiloride-sensitive cationic channels are present in the apical membrane of porcine thyroid cells in primary culture. An amiloride-sensitive (K0.5 = 150 +/- 28 nM where K0.5 is the concentration of unlabelled ligand which reduces the specific binding of the same labelled ligand by 50%) 22Na+-flux component (Km for Na+ at 18 mM) has been identified which was also blocked by the potent amiloride derivative phenamil (K0.5 = 47 +/- 21 nM). The most potent inhibitor of Na+/H+ exchange, ethylisopropyl-amiloride, hardly inhibited this 22Na+-influx component at a concentration of 21 microM. Amiloride binding sites were characterized using [3H]phenamil. The tritiated ligand binds to a single family of binding sites in thyroid membranes with a Kd value of 50 +/- 10 nM and a maximal binding capacity of 5 +/- 1 pmol/mg protein. Patch-clamp experiments have directly demonstrated the existence of a phenamil- and amiloride-sensitive cationic channel, with a conductance of 2.6 pS, which is permeable to sodium, but not very selective (PNa+/PK+ = 1.2). This channel is an important element in the regulation of the resting membrane potential of thyroid cells. PMID- 2550221 TI - Cloning and chromosomal localization of Drosophila cDNA encoding the catalytic subunit of protein phosphatase 1 alpha. High conservation between mammalian and insect sequences. AB - A 1.2-kb clone containing the full coding sequence of a protein phosphatase 1 catalytic subunit has been isolated from a Drosophila head cDNA library. It encodes a polypeptide of 302 amino acids with a molecular mass of 34.5 kDa. The predicted protein sequence is 92% identical (94% similar) to rabbit protein phosphatase 1 alpha (PP-1 alpha) demonstrating strict conservation of the phosphatase catalytic subunit over a considerable evolutionary distance. Abundant 1.6-kb and 2.5-kb mRNA transcripts were detected throughout Drosophila development. The clone hybridised to four sites on Drosophila salivary gland polytene chromosomes. The major site is at 87B6-12 on the right arm of chromosome 3. In addition, there are three secondary sites, one on the same chromosome at 96A2-5 and two on the X chromosome at 9C1-2 and 13C1-2. Isolation of a further cDNA clone, hybridising to 9C1-2 and encoding part of the catalytic subunit 88% similar to Drosophila PP-1 alpha, proves the existence of at least two transcriptionally active genes for protein phosphatase 1. PMID- 2550222 TI - Pairs of cyclic AMP analogs, that are specifically synergistic for type I and type II cAMP-dependent protein kinases, mimic thyrotropin effects on the function, differentiation expression and mitogenesis of dog thyroid cells. AB - The role of the two different isozymes of the cAMP-dependent protein kinase is still unclear. We have investigated the potential roles for each isozyme in dog thyroid cells, a model in which the function, expression of differentiation and proliferation are positively regulated by thyrotropin acting through cyclic AMP. The dog thyroid contains both type I and type II cAMP-dependent protein kinases. These isozymes were selectively activated in vitro by type-I-directed and type-II directed analog pairs. In thyroid slices, both type-I directed and type II directed analog pairs synergistically activated thyroid hormone synthesis, as measured by incorporation of 131I into proteins and thyroid hormone secretion as determined by the release of butanol-extractable 131I. In primary cultures of dog thyroid cells both isozyme-directed analog pairs synergistically enhanced iodide trapping, a marker of differentiation, and DNA synthesis, as measured by the percentage of cells incorporating [3H]thymidine into their nuclei. However, DNA synthesis was more sensitive to type-I-directed pairs. The results demonstrate that both cAMP-dependent protein kinase isozymes can mediate the action of cAMP on function, differentiation expression and cell proliferation in dog thyroid cells. PMID- 2550223 TI - Intracellular Ca2+ potentiates Na+/H+ exchange and cell differentiation induced by phorbol ester in U937 cells. AB - The human cell line U937 differentiates to monocyte macrophage-like cells in response to tumour-promoting phorbol esters. This effect is attributed to activation of protein kinase C. We show here that U937 cell differentiation induced by 12-O-tetradecanoylphorbol 13-acetate (TPA) is associated with cytoplasmic alkalinization. Ethyl-isopropyl-amiloride (EIPA), a potent inhibitor of Na+/H+ exchange, blocked both cytoplasmic alkalinization and cell differentiation. Cell acidification by addition of 2-4 mM sodium propionate also blocked TPA-induced U937 cell differentiation. These results suggest that a sustained cell alkalinization mediated by activation of Na+/H+ exchange is essential for TPA-induced differentiation in U937 cells. The increase of cytoplasmic free calcium concentration ([Ca2+]i) by addition of the calcium ionophore ionomycin enhanced TPA-induced alkalinization by increasing the apparent affinity of the Na+/H+ antiporter for intracellular H+. Treatment with ionomycin also potentiated differentiation of U937 cells induced by TPA. This synergism suggests that [Ca2+]i either potentiates the activation of protein kinase C or triggers additional transducing mechanisms. The key events of this interaction occur during the first 30 min of treatment, even though cell differentiation manifests much later. PMID- 2550224 TI - Glycosylation of glycoproteins 52 and 65 encoded by the polycythemia-inducing strain of Friend spleen focus-forming virus. Isolation of glycopeptides containing individual glycosylation sites. AB - The primary envelope gene product of the polycythemia-inducing strain of Friend spleen focus-forming virus, glycoprotein 52 (gp52), as well as its processed form, glycoprotein 65 (gp65), were isolated from virus-infected normal rat kidney cells metabolically labeled with [2-3H]mannose. Following digestion with trypsin, glycopeptides containing individual N-glycosylation sites were obtained by gel filtration and subsequent reversed-phase high-performance liquid chromatography. N-terminal amino acid sequencing of the glycopeptides demonstrated that only asparagine residues 11 and 26, located in the N-terminal domains of gp52 and gp65, carry carbohydrate substituents, while the potential N-glycosylation sites in the C-terminal portions of the molecules are not used. Carbohydrates attached were liberated by treatment with endo-beta-N-acetylglucosaminidase H or peptide: N-glycosidase F and characterized by high-performance liquid chromatography. The results demonstrated that gp52 carries similar patterns of oligomannosidic glycans in both positions. In gp65, however, asparagine residue 11 is almost exclusively substituted by complete, fucosylated N-acetyllactosaminic oligosaccharides, whereas asparagine residue 26 carries oligomannosidic or truncated N-acetyllactosaminic glycans. PMID- 2550225 TI - Effects of the cellular p53 protein on Simian-virus-40-T-antigen-catalyzed DNA unwinding in vitro. AB - It is known that large T antigen, the regulatory protein encoded by Simian virus 40 (SV40), forms tight complexes with the cellular p53 protein in SV40 transformed rodent cells. Using immunoaffinity procedures we have purified large T antigen and, in separate experiments, the cellular p53 protein. The two proteins formed complexes in vitro which bound well to double-stranded DNA fragments although in a sequence-unspecific manner. Free, uncomplexed T antigen readily converted double-stranded DNA into a single-stranded form whereas in vitro-formed p53-T-antigen complexes were inactive in this reaction. We conclude that one function of p53 in SV40-transformed mouse cells could be the inhibition of the replication initiating activity of T antigen. PMID- 2550226 TI - On the mechanism of inhibition of methanol dehydrogenase by cyclopropane-derived inhibitors. AB - Extraction of cyclopropanol-inactivated methanol dehydrogenase (MDH) gave a mixture of two interconverting compounds. The same compounds could be prepared from 2,7,9-tricarboxy-1H-pyrrolo[2,3-f]quinoline-4,5-dione (PQQ) and cyclopropanol using a metal oxide (e.g. Ag2O) as a catalyst. Structure elucidation revealed that a C5 3-propanal adduct of PQQ is formed which is present in the extract as a diastereoisomeric mixture of the ring-closed form. Cyclopropanone gave an analogous product, while cyclopropylmethanol behaved as a substrate and was oxidized by the enzyme without ring-opening. From the work described, several arguments can be derived to reject the idea that inactivation proceeds via formation of a pair of free radicals. The mechanism probably consists of a concerted proton abstraction, rearrangement of the cyclopropoxy anion to a ring-opened carbanion and attack of the latter on the electrophilic C5 of PQQ. The measured rate of inactivation (3.7 s-1) is in agreement with such a mechanism. The role of the metal oxide and the enzyme in this process is the catalysis of the addition step and possibly a positioning of the reactants. As only a sole type of quinoprotein alcohol dehydrogenase becomes inhibited, the cyclopropane derivatives studied here can be regarded as mechanism-based inhibitors. The modified PQQ in cyclopropanone-inactivated MDH is fluorescent. A fluorescent intermediate was also observed in the catalytic cycle of MDH with methanol as a substrate. Its rate of formation and decay and the strongly decreased level of fluorescence in the presence of activator are in accordance with the view that the fluorescing species is the previously found oxidized MDH.substrate (MDHox.S) complex. Since the decomposition of this complex requires activator and model studies have failed so far to mimic the enzyme, it seems that the combination of enzyme and activator is essential for the oxidation of the alcohol substrate. PMID- 2550227 TI - Modulatory activity of 9-hydroxy- and 9-hydroperoxy-octadecadienoic acid towards reactive oxygen species from guinea-pig pulmonary macrophages. AB - As guinea-pig pulmonary macrophages (PM) synthesize the linoleic acid metabolite 9-hydroxy-octadecadienoic acid (9-OH-Lin) under non-stimulated conditions in relatively large quantities, we investigated whether this product has an effect on the macrophage's own phagocytic cell function. 9-OH-Lin, and also its hydroperoxy precursor 9-hydroperoxy-octadecadienoic acid (9-OOH-Lin), influenced the generation of PM chemiluminescence, a measure of the production of reactive oxygen species. The generation of lucigenin-enhanced chemiluminescence by stimulated and non-stimulated PM was inhibited concentration-dependently. Inhibition was observed at concentrations as low as 10 nM. Since 9-OH-Lin and 9 OOH-Lin also inhibited the generation of chemiluminescence by a cell-free enzyme system, i.e. xanthine/xanthine oxidase, the inhibitory effects might represent a scavenging activity towards reactive oxygen species. 9-OH-Lin and 9-OOH-Lin did not influence other phagocytic cell functions, e.g. PM phagocytic capacity, the aggregatory response to the calcium ionophore A23187, or the release of lysosomal enzymes. The effects of 9-OH-Lin and 9-OOH-Lin could be ascribed to the hydroxy and hydroxyperoxy moiety, respectively, as evidenced by lack of effect of the native fatty acid linoleic acid. We conclude that the formation of 9-OH-Lin and 9 OOH-Lin by PM may represent a regulatory mechanism towards the cell's own activity by modulating reactive oxygen species. PMID- 2550228 TI - The role of sodium ions in methanogenesis. Formaldehyde oxidation to CO2 and 2H2 in methanogenic bacteria is coupled with primary electrogenic Na+ translocation at a stoichiometry of 2-3 Na+/CO2. AB - Cell suspensions of Methanosarcina barkeri were found to oxidize formaldehyde to CO2 and 2H2 (delta G0' = -27 kJ/mol CO2), when methanogenesis was inhibited by 2 bromoethanesulfonate. We report here that this reaction is coupled with (a) primary electrogenic Na+ translocation at a stoichiometry of 2-3 Na+/CO2, (b) with secondary H+ translocation via a Na+/H+ antiporter and (c) with ATP synthesis driven by an electrochemical proton potential. This is concluded from the following findings. Formaldehyde oxidation to CO2 and 2H2 was dependent on Na+ ions, 2-3 mol Na+/mol formaldehyde oxidized were extruded. Na+ translocation was inhibited by Na+ ionophores, but not affected by protonophores of Na+/H+ antiport inhibitors. Formaldehyde oxidation was associated with the build up of a membrane potential in the order of 100 mV (inside negative), which could be dissipated by sodium ionophores rather than by protonophores. Formaldehyde oxidation was coupled with ATP synthesis, which could be inhibited by Na+ ionophores, Na+/H+ antiport inhibitors, by protonophores and by the H+ translocating-ATP-synthase inhibitor, dicyclohexylcarbodiimide. With cell suspensions of Methanobacterium thermoautotrophicum similar results were obtained. PMID- 2550229 TI - Purification of the F420-reducing hydrogenase from Methanosarcina barkeri (strain Fusaro). AB - The 8-hydroxy-5-deazaflavin (coenzyme F420)-reducing and methyl-viologen-reducing hydrogenase of the anaerobic methanogenic archaebacterium Methanosarcina barkeri strain Fusaro has been purified 64-fold to apparent electrophoretic homogeneity. The purified enzyme had a final specific activity of 11.5 mumol coenzyme F420 reduced.min-1.mg protein-1 and the yield was 4.8% of the initial deazaflavin reducing activity. The hydrogenase exists in two forms with molecular masses of approximately 845 kDa and 198 kDa. Both forms reduce coenzyme F420 and methyl viologen and are apparently composed of the same three subunits with molecular masses of 48 kDa (alpha), 33 kDa (beta) and 30 kDa (gamma). The aerobically purified enzyme was catalytically inactive. Conditions for anaerobic reductive activation in the presence of hydrogen, 2-mercaptoethanol and KCl or methyl viologen were found to yield maximal hydrogenase activity. Determination of the apparent Km of coenzyme F420 and methyl viologen gave values of 25 microM and 3.3 mM, respectively. The respective turnover numbers of the high molecular mass form of the hydrogenase are 353 s-1 and 9226 s-1. PMID- 2550230 TI - Carboxylic acid reductase: a new tungsten enzyme catalyses the reduction of non activated carboxylic acids to aldehydes. AB - An enzyme which we call carboxylic acid reductase (aldehyde dehydrogenase) seems to be the first which is able to reduce non-activated carboxylic acids to aldehydes at the expense of reduced viologens. There is no further reduction of the aldehydes to the corresponding alcohols. In the presence of oxidized viologens aldehydes can be dehydrogenated to carboxylic acids roughly 20 times faster than the latter are reduced. The specific enzyme activity in crude extracts is about 100 times increased if 10 microM tungstate and a sulphur source in addition to sulphate is given to the growth medium of Clostridium thermoaceticum. Carboxylic acid reductase seems to be present in two forms. One has an apparent molecular mass of about 240 kDa and is bound to red-Sepharose, whereas, the other, a form of an apparent molecular mass of about 60 kDa, is not bound. SDS gel electrophoresis shows a higher complexity. The very labile enzyme has been enriched by a factor of about 145 by binding to octyl-Sepharose and further chromatographic separation by red-Sepharose and FPLC using Mono-Q and phenyl-Superose columns. After cell growth in the presence of [185W]tungstate, radioactivity coincides with the two forms of enzyme activity during all purification steps. This is also the case when the enzyme is electrophoretically separated on polyacrylamide slab gels. PMID- 2550231 TI - Detection of cytomegalovirus by the early-antigen immunofluorescence test versus conventional tissue culture. AB - The two methods commonly used to diagnose cytomegalovirus (CMV) infections, conventional tissue culture and detection of early CMV nuclear antigen by immunofluorescence from cell culture, were performed in parallel on 597 clinical specimens. CMV was detected by the early-antigen test in 108 samples, of which 102 (94%) were detected 1 to 3 days after inoculation. Of these 108 CMV-positive specimens, seven were negative on conventional culture. Two samples negative in the early-antigen test were positive on conventional culture. Thus, CMV was detected in 110 specimens. A cytopathic effect in conventional tissue culture occurred 9 to 42 days after inoculation. The diagnosis of CMV infection was possible by the conventional method 29.6 +/- 12.7 days and by early-antigen immunofluorescence 1.9 +/- 1.5 days after obtaining the specimen. The rapid early antigen test was slightly more sensitive than culture, and fewer samples were lost due to bacterial or fungal infections during incubation. Detection of CMV by conventional culture usually requires several weeks and provides a diagnosis only retrospectively. The main advantage of the early-antigen test is that a virologically proven diagnosis of CMV infection is available at an early stage. PMID- 2550232 TI - Detection of Bordetella pertussis by determination of adenylate cyclase activity. AB - The adenylate cyclase activity of Bordetella pertussis in clinical isolates was measured in calmodulin-supplemented Stainer-Scholte broth by the rate of conversion of ATP to cyclic AMP. Analysis of 250 stock strains of Bordetella pertussis showed that measurable adenylate cyclase activity was produced by all strains. In clinical tests Bordetella pertussis was isolated from 135 (22%) of 605 swab samples. Increased adenylate cyclase activity was detected in 124 (92%) Stainer-Scholte broth cultures of these samples. A total of 475 swabs contained other bacteria or had no growth; only one of the Stainer-Scholte broth cultures of these swab samples contained measurable adenylate cyclase activity. The results indicate that testing for adenylate cyclase activity provides a specific and sensitive means for detecting Bordetella pertussis in clinical specimens. PMID- 2550233 TI - Comparative antimicrobial activity of the new macrolides against Borrelia burgdorferi. AB - The in vitro and in vivo activity of the new macrolides azithromycin, clarithromycin and roxythromycin was compared with that of erythromycin against Borrelia burgdorferi. In in vitro tests using ten clinical isolates all macrolides were highly active against Borrelia burgdorferi (MIC90 0.015-0.06 micrograms/ml). Azithromycin was more potent than the other macrolides in experimental animal infection, eradicating the organism in all animals tested at a dosage of 8 mg/kg. PMID- 2550234 TI - Activity of new macrolides against Bordetella pertussis and Bordetella parapertussis. AB - MICs and MBCs of four new macrolides (azithromycin, clarithromycin, dirithromycin and roxithromycin) and two older macrolides (erythromycin and josamycin) for Bordetella pertussis and Bordetella parapertussis were determined. The activity of the new macrolides was as good as that of erythromycin, while josamycin was slightly less active. Bordetella parapertussis was more resistant than Bordetella pertussis. PMID- 2550236 TI - Comparative in vitro activity of daptomycin (LY146032) and vancomycin against gram-positive cocci determined using a pharmacokinetic model. AB - The activity of daptomycin (LY146032) and vancomycin was compared for methicillin sensitive and methicillin-resistant (MRSA) strains of Staphylococcus aureus and for Enterococcus faecalis, using a semi-automated model that allows examination of time-kill curves with diminishing drug concentrations, thus reflecting in vivo pharmacokinetics. Exposure to daptomycin resulted in rapid killing of all strains of staphylococci and Enterococcus faecalis tested. Methicillin-sensitive staphylococcal strains appeared marginally more susceptible than MRSA. By contrast, the activity of vancomycin appeared much reduced. For all strains, greater than 90% kill was not observed within 24 h. Enterococcus faecalis was uniformly less susceptible to both agents than were staphylococci. The results suggest that despite similar values for MICs, the activity of daptomycin against strains of Staphylococcus aureus and Enterococcus faecalis is comparable to and may exceed that of vancomycin, although the significance of such differences is uncertain. PMID- 2550237 TI - The viral lesion (HPV) of the lower genital tract in teen agers: clinical and therapeutic aspects. AB - From January 1985 to July 1987, 116 adolescents affected by the lesions of the lower genital tract (Human Papilloma Virus +/- Intraepithelial Neoplasia) came to the laser surgery service of the 2nd Department of the Obstetric and Gynecologic Clinic of the University of Rome "La Sapienza" for the treatment of the lesions. The patients were examined also from the epidemiologic point of view and sides effects of the treatment were evaluated. A cure rate of 92% is certainly a significant value, especially considering the higher frequency of recurrence at the vulvovaginal level. PMID- 2550235 TI - Persistent herpes simplex virus infection and mechanisms of virus drug resistance. AB - Herpes simplex virus (HSV) is susceptible to a variety of antiviral compounds, most of which are nucleoside analogues that interfere with DNA metabolism involving the virus enzymes DNA-polymerase and thymidine kinase. Single mutations in the virus genome give rise to resistant mutants following selection in vitro in the presence of a particular drug, and in this respect HSV is similar to several other viruses. Such mutants have been invaluable research tools. HSV is responsible for a variety of lesions which tend to be recurrent, owing to the special ability of the virus to remain latent in and reactivate from neural tissue. The consequences of this upon clinical resistance are discussed in the present review. In fact, clinical resistance in HSV infections has not yet become widespread but does appear to be especially important in immunocompromised patients, including those suffering from AIDS. HSV is proposed as an important model for the investigation of drug resistance in other, more complex organisms, and with respect to antiviral strategies against the human immunodeficiency virus. PMID- 2550238 TI - HLA typing in familial testicular cancer. AB - Support is given to the theory of genetic influence in the etiology and pathogenesis of testicular germ cell tumors by the report of four new familial clusters of testicular cancer. One pair of brothers both afflicted with seminoma underwent HLA typing and they were shown to share one complete haplotype. A review of 119 families of the literature with aggregation of germ cell tumors is given. The evaluation of eight previously reported familial clusters of germ cell tumors with HLA-typing results in conjunction with the present case shows a haplotype sharing in all cases which is more than expected. This finding is another piece of evidence for the theory of genetic anticipation of germ cell tumors. PMID- 2550239 TI - Malignant fibrous histiocytoma of the kidney. AB - Malignant fibrous histiocytoma is a primitive mesenchymal tumor which seldom occurs in the genitourinary organs. Despite radical operation, prognosis is generally poor and seems not to be improved by chemotherapy or radiotherapy. We present a case of a malignant fibrous histiocytoma of the renal capsule. The tumor could be removed totally and no signs of metastases were found. Eight months later the patient was readmitted with recurrence of tumor and widespread metastases. PMID- 2550240 TI - Receptor signalling and intracellular calcium in neutrophil activation. PMID- 2550241 TI - Oestrogen-induced changes in lipoprotein metabolism: role in prevention of atherosclerosis in the cholesterol-fed rabbit. AB - Administration of moderate pharmacological doses of oestrogen to cholesterol-fed rabbits for 12 weeks resulted in a dramatically retarded development of arterial lesions as compared to non-oestrogen-treated, cholesterol-fed rabbits (7% vs. 47% aortal involvement). Oestrogen treatment was associated with a retarded increase in plasma cholesterol, and five times higher high density lipoprotein (HDL) to very low density lipoprotein (VLDL) cholesterol ratio. Expression of hepatic lipoprotein receptor activity, as detected by heparin-releasable binding of 125I hypercholesterolaemic VLDL, was heavily suppressed by cholesterol feeding. Administration of oestrogen modulated this response and resulted in higher receptor expression. In accordance, oestrogen treatment resulted in a less prominent reduction of 125I-hypercholesterolaemic VLDL clearance in the cholesterol-fed rabbits. VLDL from both groups of cholesterol-fed animals stimulated cholesteryl ester synthesis in cultured macrophages to the same extent. Thus, in rabbits under a dietary cholesterol load, oestrogen counteracted hepatic lipoprotein receptor suppression, reduced plasma VLDL- and increased plasma HDL-cholesterol levels, and to a large extent abolished the development of atherosclerosis. PMID- 2550242 TI - Leukotriene B4 metabolism in neutrophils of patients with chronic granulomatous disease: phorbol myristate acetate decreases endogenous leukotriene B4 via NADPH oxidase-dependent mechanism. AB - We studied the effect of phorbol myristate acetate (PMA) on endogenous leukotriene B4 (LTB4) metabolism of calcium ionophore A23187-stimulated human neutrophils. Preincubation of normal neutrophils with PMA significantly suppressed the recovery of endogenous LTB4 induced by A23187. PMA did not suppress the recovery of LTB4 produced by neutrophils from patients with chronic granulomatous disease (CGD), which is known to be defective in NADPH oxidase activation to produce reactive oxygen species (ROS). PMA inhibited the formation of omega-oxidation products of LTB4, but enhanced arachidonic acid release in normal and CGD neutrophils. Furthermore, 5-lipoxygenase activity of 10,000 x g supernatants from normal neutrophils pretreated with PMA was equivalent to that of the controls. Decrease in LTB4 recovery was not attributed to the suppression of the intracellular Ca2+ increase. Thus, it is suggested that reactive oxygen species (ROS) produced by PMA may directly affect endogenous LTB4 and convert it into metabolite(s) distinct from omega-oxidation products. PMID- 2550243 TI - Acute calcium entry blockade inhibits the blood pressure but not the hormonal responses to angiotensin II. AB - The effects of acute calcium entry blockade by isradipine (IS) and placebo (P) on the haemodynamic and humoral responses to angiotensin II (A II) have been compared in two groups of 9 patients with essential hypertension. During 4 sequential periods each of 20 min, an i.v. infusion of A II 0, 2, 4 and 8 ng.kg 1.min-1 was given before (control) and 30 min after the oral administration either of IS or P. After IS, both the blood pressure and the angiotensin II induced pressor effect were significantly reduced. Isradipine increased the heart rate and this cardio-acceleration was potentiated by A II. In contrast, when A II was infused in the absence of IS, heart rate tended to decrease. IS stimulated plasma renin activity and reduced plasma aldosterone. However, it did not affect either the inhibition of plasma renin activity or the rise in plasma aldosterone in response to A II. In conclusion, acute calcium entry blockade in patients with essential hypertension reduces the pressor response to A II, but not the A II induced inhibition of renin and increase in plasma aldosterone. PMID- 2550244 TI - Effects of prolonged administration of the angiotensin converting enzyme inhibitor CGS 16617 in normotensive volunteers. AB - A new, orally active angiotensin converting enzyme (ACE) inhibitor, CGS 16617, has been evaluated in normotensive subjects during acute and prolonged administration. Single ascending doses of CGS 16617 20 to 100 mg were given to 9 normotensive volunteers at one week intervals and the changes in blood pressure, plasma ACE and renin activity were examined up to 72 h after drug intake. Also, CGS 16617 50 mg/day or placebo were given for 30 days to 8 and 6 normotensive subjects, respectively, maintained on an unrestricted salt diet. Blood pressure was measured daily in the office and ambulatory blood pressure profiles were also obtained before, during and after therapy, using the Remler M 2000 blood pressure recording system. CGS 16617 was an effective and long lasting ACE inhibitor. It did not induce a consistent change in blood pressure, but, the individual responses were very variable and several subjects experienced a clear decrease in the average of the blood pressures recorded during the daytime. PMID- 2550246 TI - Fc gamma RII expression in resting and activated B lymphocytes. AB - In this report, we analyze the expression of the type II receptor for the Fc region of IgG (Fc gamma RII) in resting and lipopolysaccharide (LPS)-activated murine B lymphocytes. Fc gamma RII is encoded by two genes, alpha and beta. The beta gene encodes two mRNA, beta 1 and beta 2, which are generated by alternative splicing. Using an S1 nuclease protection assay, we found that resting and activated B lymphocytes express predominantly the beta 1 transcript. Very low levels of the beta 2 mRNA were detected in this assay, while no expression of the alpha transcript could be detected. Quantitative Northern blot analysis showed that the amount of Fc gamma RII beta mRNA was increased 9-fold in LPS-activated B lymphocytes. The expression of Fc gamma RII during the various phases of B cell activation was then studied by immunofluorescence using the monoclonal antibody 2.4G2. LPS stimulation induced an increase of the Fc gamma RII cellular pool as well as of its expression at the surface of B lymphocytes. The rise in Fc gamma RII surface expression occurred after the induction of class II antigens (Ia) and before transferrin receptor induction. Fc gamma RII expression was found to be enhanced during the G1 phase of the cell cycle since (a) only large cells (i.e. those that had entered the G1 phase) expressed an increased amount of Fc gamma RII and (b) blocking the entry of activated cells into the S phase (with the ion channel blocker quinine) did not affect the Fc gamma RII induction by LPS. Furthermore, only B cell activators that induced cells to enter into G1 [LPS and F(ab')2 anti-IgM antibodies, but not interleukin 4] caused an increase in the expression of Fc gamma RII. These results show that the increase in the membrane expression of Fc gamma RII occurs during the early G1 phase, establishing it as a marker for the entry of B lymphocytes into the cell cycle. PMID- 2550245 TI - Effect of human atrial natriuretic peptide on blood glucose concentrations and hormone stimulation during insulin-induced hypoglycaemia in healthy man. AB - A double-blind placebo-controlled study using the double-dummy technique has been done to examine whether the responses of pituitary and adrenal hormones to insulin-induced hypoglycaemia were impaired by a pharmacological dose of human atrial natriuretic peptide (human ANF-(99-126),hANP). After an overnight fast eight male healthy volunteers (aged 23-40 years) received in random order i.v. bolus injections of insulin 0.125 U.kg-1 + placebo,hANP 100 micrograms + placebo,insulin + hANP, or both placebo preparations. In the insulin-only experiment, human growth hormone, adrenocorticotrophic hormone, cortisol, aldosterone, plasma renin activity, adrenaline, and noradrenaline were all stimulated by hypoglycaemia. In the hANP-only experiment there were no hormonal changes other than decreases in plasma renin activity and aldosterone concentration. In the insulin + hANP experiment the nadir of blood glucose was decreased to 1.3 from the 2.0 mmol.1-1 found in the insulin-only experiment. The exaggerated hypoglycaemia resulted in increased stimulation of human growth hormone, adrenocorticotrophic hormone and adrenaline when compared to the insulin only experiment. The rise in the cortisol and aldosterone concentrations was only slightly increased, and the stimulation of plasma renin activity was blunted. Unexpectedly, hANP was found to enhance the hypoglycaemic action of insulin, most probably by inhibiting insulin degradation within the liver. There was no evidence of an inhibitory effect of hANP on the stimulation of pituitary or adrenal hormones during insulin-induced hypoglycaemia. The reduction in renin may indicate an inhibitory action of hANP on catecholaminergic effects within the kidney. PMID- 2550247 TI - Heat shock protects WEHI-164 target cells from the cytolysis by tumor necrosis factors alpha and beta. AB - Elevated temperatures and a number of other types of stress induce synthesis of a small number of highly conserved proteins, the heat shock proteins, in a wide variety of cells. The structure and regulation of these proteins have been intensively studied but the question of the function of this universal response has remained unanswered. We studied the effect of heat shock on tumor necrosis factor-alpha (TNF-alpha)- and -beta (TNF-beta)-mediated cytolysis of WEHI-164 clone 13 target cells. One hour pretreatment of target cells at 42 degrees C decreased rTNF-alpha-mediated lysis by 65.3%, 50.5% and 44.8% and TNF-beta mediated lysis by 61.9%, 43.2% and 38.9% at cytokine concentrations of 0.5 ng/ml, 5 ng/ml and 50 ng/ml, respectively, in an 18-h Cr-release assay. The effect was maximal when TNF-alpha was added 1 h after the heat shock and then gradually declined, being almost undetectable after 2 days. This pattern was found to roughly coincide with the kinetics of hsp68, the major heat-induced protein in murine cells. Heat shock treatment had no protective effect when given 1 h after addition of recombinant TNF-alpha. The heat-induced target cell resistance was not associated with decreased binding of recombinant TNF-alpha to its receptor. Inhibition of protein synthesis by cycloheximide diminished this effect by 76% and inhibition of transcription by actinomycin D abolished it completely, suggesting that de novo synthesized heat-induced proteins protect target cells from TNF-mediated lysis in heat shock-treated WEHI cells. PMID- 2550248 TI - Inhibition by chlorpromazine of lymphokine-specific mRNA expression in human thymocytes. AB - This study was designed to determine the effect of the phenothiazine chlorpromazine (CPZ) on the activation of human thymocytes. We provide evidence that CPZ inhibits the accumulation of mRNA specific for the lymphokines, interleukin 2, interferon-gamma, tumor necrosis factor alpha and the proto oncogene c-myc; by contrast, the accumulation of mRNA specific for the alpha chain of the interleukin 2 receptor and the subsequent early expression of Tac antigen on the cell surface is not inhibited by CPZ. The inhibition of the expression of lymphokine-specific mRNA results in a decrease in interferon-gamma synthesis and in inhibition of thymocyte proliferation as determined by the incorporation of [3H]thymidine. In addition, we show that activation of protein kinase C (PKC) in human thymocytes by 12-O-tetradecanoyl phorbol 13-acetate (TPA) causes the phosphorylation of a protein of a molecular mass of approximately 75 kDa. The function of this protein is as yet not defined, but it is possible that it plays a role in the transduction of the signals to the nucleus which in turn elicit the expression of the genes coding for c-myc and for the lymphokines required for thymocyte activation. We also demonstrate that CPZ, like the immunosuppressant drug cyclosporin A does not inhibit the phosphorylation of the 75-kDa protein which is induced by the activation of PKC by TPA and does not affect phosphoinositide breakdown, indicating that it exerts its effect at a site distal to the activation of PKC. These observations demonstrate that CPZ has an immunoregulatory function in addition to its psychotropic activity. PMID- 2550249 TI - Analysis of T cell receptor gamma chains from adult CD4- CD8- thymocytes. AB - The role of T cell receptor (TcR) gamma genes in T cell development has not been determined. To extend our understanding of the repertoire of TcR gamma expression, we prepared a cDNA library from CD4-CD8- adult BALB/c thymocytes and cloned and sequenced 15 TcR gamma genes from this cDNA library. We found that two clones were transcripts of the unrearranged C gamma 2 gene and that three clones terminated in the J gamma 2 region. Nine of the remaining clones were V gamma 1.2 J gamma 2 C gamma 2 genes and five of these were in frame. Only one clone corresponded to C gamma 1 and resulted from an in-frame V gamma 2 J gamma 1 C gamma 1 join. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the gamma chain proteins from the surface of both BALB/c and C57BL/6 adult CD4 CD8- thymocytes did not detect the 32-kDa V gamma 1.2 C gamma 2 protein, but did detect the 35-kDa V gamma C gamma 1 protein. These results suggest that despite the abundance of full-length functional V gamma 1.2 C gamma 2 transcripts in the thymocyte subset, the protein product is not expressed on the cell surface as the predicted 32-kDa gamma protein. Finally, our analysis of the V-J joining of the gamma genes reveals both flexibility at the V-J junction and extensive N-region nucleotide addition that lead to diversity of the predicted protein sequence. PMID- 2550250 TI - Tissue selectivity of endothelin. AB - The effects of endothelin, a potent endogenous vasoconstrictor peptide, were examined in a range of vascular and non-vascular tissues. At concentrations that cause vasoconstriction in portal vein and aorta, the peptide strongly contracted rat uterus, trachea and vas deferens, but not guinea pig ileum. Nifedipine, a dihydropyridine calcium anatgonist, partially inhibited these contractions. Endothelin had no inotropic or chronotropic effect on the isolated rat heart. The peptide did not modulate secretion at the neuromuscular junction, from adrenal medullary cells or neutrophils, nor affect secretion or aggregation of platelets. The tissue responsiveness to endothelin was not the same as the tissue distribution of dihydropyridine receptors. This supports the idea that endothelin interacts with a specific receptor distinct from dihydropyridine sensitive calcium channels. The contractile effect of endothelin on non vascular smooth muscle suggests that the concept of endothelium dependent modulation of vascular smooth muscle tone may be extended to include epithelium dependent modulation of non vascular tissues. PMID- 2550251 TI - Beneficial effects of a specific leukotriene receptor antagonist in splanchnic artery occlusion shock. AB - The purpose of this study was to examine the effects of a new potent peptidoleukotriene receptor antagonist, SK&F 104353, in splanchnic artery occlusion shock. SK&F 104353 was administered as a 1 mg/kg initial bolus followed by an infusion of 3 mg/kg per h for the entire 2 h post-reperfusion observation period. In a group of conscious rats, this dose of SK&F 104353 shifted the LTD4 dose response curve rightward 10-fold, indicating effective antagonism of peptidoleukotriene responses in the rat. Anesthetized rats subjected to splanchnic artery occlusion shock survived an average of only 98 +/- 8 min whereas all animals receiving SK&F 104353 survived the 2 h reperfusion period (P less than 0.02 from vehicle). Therefore, the survival rate of the splanchnic artery occlusion shock group of rats receiving SK&F 104353 was improved to 100% compared with 50% survival for the vehicle-treated splanchnic artery occlusion shock group (P less than 0.025). In the splanchnic artery occlusion shock + SK&F 104353 group the increase in the plasma activities of the lysosomal hydrolase, cathepsin D, and the cardiotoxic peptide, myocardial depressant factor, were significantly attenuated in comparison to the splanchnic artery occlusion shock + vehicle group (P less than 0.025). These data indicate that the peptidoleukotriene receptor antagonist, SK&F 104353 is beneficial in splanchnic artery occlusion shock, and furthermore suggests that it may be a therapeutically useful agent in bowel ischemic shock. PMID- 2550252 TI - Sertraline potently displaces (+)-[3H]3-PPP binding to sigma sites in rat brain. PMID- 2550253 TI - The behavioural effects of MK-801: a comparison with antagonists acting non competitively and competitively at the NMDA receptor. AB - The selective non-competitive NMDA receptor antagonist, MK-801, potently blocked convulsions induced in the mouse by N-methyl-DL-aspartic acid (NMDLA) with an i.v. ED50 dose of 0.2 mg/kg. Similar doses of MK-801 were also effective in blocking seizures induced by pentylenetetrazol (PTZ), electroshock and by sound in audiogenic seizure-prone animals. Other less selective non-competitive NMDA receptor antagonists including phencyclidine (PCP), thienylcyclohexylpiperidine (TCP), (+)-N-allylnormetazocine [+)-NANM, (+)-SKF 10,047) and ketamine also blocked NMDLA-induced seizures with a rank order of potency of MK-801 greater than PCP greater than TCP = (+)-NANM greater than ketamine. The competitive NMDA receptor antagonist, 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP) blocked NMDLA-induced seizures with an ED50 of 4.5 mg/kg, 22- and 560-fold more potently than the competitive antagonists, 2-DL-amino-7-phosphonoheptanoic acid (2-APH) and 2-DL-amino-5-phosphonovaleric acid (2-APV), respectively. MK-801 was the most potent of the non-competitive antagonists to induce a motor syndrome including head weaving, body rolling, increased locomotion and ataxia, characteristic of the behavioural response to PCP in the mouse. The syndrome was also present following injection of the competitive NMDA receptor antagonists, although they were generally less potent (probably a reflection of poor brain penetration) and less efficacious than the non-competitive antagonists. For all compounds except CPP, the anticonvulsant ED50 dose was close to the minimum effective dose to induce motor stimulation: CPP was 5- to 10-fold more potent as an anticonvulsant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550254 TI - Heterogeneity of the effects of monensin on two types of contractions evoked by inhibition of Na+-K+ ATPase in the guinea-pig vas deferens. AB - The interactions between ouabain or K+-free medium and monensin on noradrenaline (NA) release and Ca2+-evoked contractions were assessed in guinea-pig vas deferens. Ouabain (10 microM) produced large, sustained contractions and monensin (1 microM) small, transient contractions. The ouabain-evoked contraction tended to be potentiated in the presence of monensin. The marked contraction induced by ouabain plus monensin was nearly abolished by phentolamine or by treatment with 6 hydroxydopamine. Ouabain caused the release of NA from the tissue, as determined in an HPCL-ECD study. This ouabain-evoked release of NA was enhanced by the simultaneous administration of monensin. High Ca2+ (10 mM) per se, ouabain, monensin or K+-free medium did not elicit contraction of the tissue in the presence of phentolamine. However, 10 mM Ca2+ caused a large contraction 45-65 min after exposure to ouabain or K+-free medium and this contraction was suppressed by monensin. The contraction evoked by Ca2+ in the presence of ouabain was further inhibited by amiloride, an inhibitor of Na+-Ca2+ or Na+-H+ exchange transport, but not by nifedipine, a voltage-dependent Ca2+ antagonist.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550255 TI - Ethanol-induced bronchodilatation in TEA-treated canine tracheal smooth muscle is mediated by a beta-adrenoceptor-dependent mechanism. AB - The effects of moderate concentrations of ethanol (8-34 mM) on the electromechanical activity of airway smooth muscle cells of canine trachealis, stimulated by the spasmogen tetraethylammonium (TEA), are described for in vitro and cultured reaggregate preparations. Ethanol produced a concentration-dependent hyperpolarization, and suppression of action potentials in smooth muscle preparations, in vitro, whereas it was without effect in cultured airway smooth muscle cells. In the presence of the beta-adrenoceptor antagonist propranolol (1 microM), ethanol had no effect on in vitro preparations. Isoproterenol (0.1 microM) produced hyperpolarization and suppression of action potentials in airway smooth muscle of both preparations. These effects were not observed when propranolol was additionally present. This suggests that both in vitro, and cultured airway smooth muscle preparations maintained their beta-receptors, and that ethanol caused the release of endogenous catecholamine from adrenergic nerve endings which apparently remained intact in in vitro, but not in cultured airway smooth muscle preparations. PMID- 2550256 TI - Presynaptic alpha 2-adrenoceptors modulate the release of [3H]noradrenaline from rat spinal cord dorsal horn neurones. AB - Slices of the dorsal half of the rat spinal cord were used to investigate the existence of a noradrenergic feedback modulation of noradrenaline release. After crude preparation of the vertebral column, the spinal cord was ejected by hydraulic pressure and transverse slices were cut. These were preincubated with [3H]noradrenaline during 0.1 Hz electrical stimulation and then superfused and stimulated electrically for two periods. The stimulation-evoked release of [3H]noradrenaline was Ca2+-dependent and tetrodotoxin-sensitive. Pretreatment of the animals with the noradrenergic neurotoxin, DSP-4, reduced the tritium content in the slices and the stimulation-evoked release to less than 10% of the controls. Clonidine (0.01-1 microM) inhibited the evoked overflow by 60% maximally and yohimbine (0.1-1 microM) enhanced it by 160% maximally. The effects of clonidine were antagonized by yohimbine. These results provide evidence that noradrenaline release from spinal cord slices is controlled by an alpha 2 adrenoceptor-mediated, negative feedback mechanism. PMID- 2550257 TI - 5 alpha-pregnan-3 alpha,20 alpha-diol behaves like a partial agonist in the modulation of GABA-stimulated chloride ion uptake by synaptoneurosomes. AB - In rat cortical synaptoneurosomes, the maximum potentiation of GABA-stimulated 36Cl uptake produced by 5 alpha-pregnan-3 alpha,20 alpha-diol (5 alpha pregnanediol) is significantly less than that elicited by 5 alpha-pregnan-3 alpha ol-20-one (3 alpha-OH-DHP). This observation suggests that 5 alpha-pregnanediol may be a partial agonist whereas 3 alpha-OH-DHP acts as a full agonist at a common site on or near the GABAA/benzodiazepine receptor-chloride ionophore complex (GBRC). This hypothesis is supported by the finding that 5 alpha pregnanediol will antagonize in a dose-dependent manner the enhancement of GABA stimulated 36Cl uptake produced by 3 alpha-OH-DHP under certain conditions. Collectively, these findings support the hypothesis that GBRC-active progesterone metabolites with varying degrees of efficacy exist as reflected by their differential ability to potentiate 36Cl uptake in brain synaptoneurosomes. PMID- 2550258 TI - Serotonergic involvement in the inhibitory effects of repeated buspirone treatment on synaptic transmission in the hippocampus. AB - The effects of acute and long-term treatment with buspirone on synaptic transmission in the hippocampus were compared in alert rats with chronic indwelling electrodes and cannula. Buspirone produced a transient dose-dependent reduction in the amplitude of the excitatory postsynaptic potential (EPSP) when acutely injected either systemically (0.3-3.0 mg/kg i.p.) or directly into the hippocampus (0.1-1.0 microgram i.h.). Whereas acute application of 0.5 mg/kg i.p. produced a 20% reduction which reversed within 2 h, during long-term treatment with this relatively low dose there was a gradual reduction of the baseline EPSP amplitude which reached a maximum (40%) between days 7-14 and which did not reverse completely until 72 h after the last injection. Intrahippocampal injection of either buspirone or 5-hydroxytryptamine did not have any additional effect during the period of baseline reduction. The 5-HT1A receptor antagonist spiroxatrine (1 mg/kg i.p.) produced a transient reversal of the effect of chronic buspirone. It is concluded that the chronic inhibitory effect of buspirone is probably an extension of its acute action on 5-HT1A receptors in the hippocampus. PMID- 2550259 TI - The role of alpha- and beta-adrenoceptors in the response to noradrenaline of lymphatic vessels isolated from the bovine mesentery. AB - The role of postsynaptic adrenoceptors in the spontaneous activity of isolated bovine mesenteric lymphatic vessels was characterized. Low concentrations of noradrenaline (less than 10(-6) M) increased the amplitude and the frequency of the spontaneous contractions whereas high concentrations (less than 10(-6) M) depressed the activity. beta-Adrenoceptor blockade with propranolol enhanced the stimulation by noradrenaline and suppressed the inhibition. alpha 1-Blockade by prazosin or alpha 2-blockade by idazoxan antagonized the stimulation by noradrenaline and potentiated the inhibition. Activation of alpha 1-adrenoceptors by methoxamine, or of alpha 2-adrenoceptors by clonidine, stimulated lymphatic activity. The increases in the amplitude and frequency of the spontaneous contractions produced by each agonist were competitively antagonized by prazosin and idazoxan, respectively. Our results confirm the presence of postjunctional beta-, alpha 1-and alpha 2-adrenoceptors in lymphatic vessels and show that each of these receptors plays an important role in the adrenergic regulation of the spontaneous activity, with beta-adrenoceptors mediating inhibition, and alpha 1- and alpha 2-adrenoceptors mediating stimulation. PMID- 2550260 TI - Effects of benzodiazepines on serotonin release from rat mast cells. AB - Ro5-4864, diazepam and chlordiazepoxide inhibited the concanavalin A-induced [14C]serotonin release from rat mast cells dose dependently with IC30 values 38, 115 and 160 microM, respectively. They also inhibited concanavalin A-induced 45Ca uptake, with IC50 values 180, 860 and 1800 microM, respectively. Clonazepam slightly inhibited serotonin release, but medazepam did not, and neither compound inhibited the calcium uptake stimulated by concanavalin A. The potencies of benzodiazepines to inhibit concanavalin A-induced serotonin release and 45Ca uptake were correlated with their binding affinities to the peripheral type of benzodiazepine binding sites. At higher concentrations, these benzodiazepines caused release of both serotonin and lactate dehydrogenase, due to their cytotoxicity. The calcium channels of mast cells are probably not voltage dependent, as the agonists of voltage-dependent calcium channels, Bay k 8644 and YC-170, did not stimulate serotonin release. Moreover, Ro5-4864, diazepam and chlordiazepoxide inhibited A23187-induced serotonin release. Mast cells have high contents of calmodulin (602 +/- 20 ng/10(6) cells), and benzodiazepines inhibited calmodulin. The benzodiazepine inhibitory effects on the serotonin release induced by A23187 seemed to be partly due to their calmodulin-inhibiting activities. These results suggest that inhibition of serotonin release by benzodiazepines in mast cells activated by concanavalin A is mainly due to inhibition of calcium channels, which may be controlled by the peripheral type of benzodiazepine binding sites. PMID- 2550261 TI - Ten day administration of desipramine produces an increase in KL/KH for beta receptors in rat hippocampus. AB - The action of chronic desipramine administration on agonist and antagonist binding to beta-adrenoceptors was investigated in rat frontal cortex and hippocampus. Ten day treatment with desipramine (10 mg/kg per day i.p.) resulted in a significant 34% decrease in the density of beta-adrenoceptors in membranes from rat frontal cortex but not hippocampus. The KD values for antagonist (iodocyanopindolol) did not change in either tissue after the desipramine treatment. Desipramine had no effect on the affinity of the agonist isoproterenol in frontal cortex. The percentage of receptors in the high affinity state for isoproterenol (% RH) decreased by 11% in the membranes from frontal cortex after desipramine administration. In contrast, in hippocampal membranes, desipramine treatment produced a decrease in KH (dissociation constant for isoproterenol binding to the high affinity state) and consequently a 137% increase in the ratio of dissociation constants for isoproterenol to the low and high affinity conformations (KL/KH; an putative index of coupling). This observation is new and suggests possible supercoupling of beta-adrenoceptors in hippocampus during desipramine treatment. Thus, in addition to desensitization of beta-adrenoceptors in rat frontal cortex ten day administration of desipramine causes a change compatible with supercoupling of beta-receptors in hippocampal membranes. PMID- 2550262 TI - Molecular target size analyses of the NMDA-receptor complex in rat cortex. AB - The molecular weights of different subunits of the NMDA-receptor complex were determined by high-energy radiation inactivation analyses of the binding of [3H]L glutamate, [3H](3-(+/-)-2-(carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP), [3H]N-(1-[2-thienyl]cyclohexyl)3,4-piperidine (TCP) and [3H]glycine to rat cortical membranes. The molecular target sizes of [3H]L-glutamate binding (the recognition site), [3H]TCP binding (the ionophore) and [3H]glycine (a modulatory unit) were similar: 121,000, 118,000 and 115,000 Da, respectively. These results suggest that the three subunits are on the same protein. The molecular weight of [3H]CPP binding was 209,000 Da. This suggests that in order to bind [3H]CPP (a competitive antagonist) with high affinity an additional macromolecule may be associated to the agonist site. PMID- 2550263 TI - In vivo pertussis toxin treatment attenuates some, but not all, adenosine A1 effects in slices of the rat hippocampus. AB - In order to examine the involvement of G-proteins in mediating the different effects of adenosine A1-receptor stimulation in rat hippocampus we injected pertussis toxin (PTX) intraventricularly close to the hippocampus and examined its effect in slices 48-60 h later. The in vivo PTX treatment caused a partial (50 +/- 5%) inhibition of the [32P]ADP ribosylation produced by PTX added together with [32P]NAD in vitro. Such PTX treatment eliminated the electrophysiologically determined gamma-amino-n-butyric acid (GABA)B receptor response in the hippocampal CA1 region, but GABAA effects were unaffected. The adenosine (50 microM)-mediated hyperpolarization and decrease in input resistance as well as the adenosine-mediated inhibition of low calcium-induced bursting in pyramidal CA1 neurons were virtually abolished. The same was true for the decrease in [3H]cyclic AMP accumulation that is produced by the adenosine analogue R-N6-phenylisopropyl adenosine (R-PIA) in forskolin-treated hippocampal slices. As far as modulation of transmitter release was concerned, the R-PIA (1 microM)-induced inhibition of release of both [3H]noradrenaline (NA) and [3H]acetylcholine (ACh) evoked by field stimulation in hippocampal slices was affected hardly or not at all by pertussis toxin treatment. The inhibitory effect of adenosine on field excitatory postsynaptic potential (EPSP)s evoked in the CA1 region was unaltered by PTX pretreatment. The present results show that in vivo pertussis toxin treatment can inhibit some but not all A1-adenosine-receptor effects. This strongly suggests that closely similar A1 receptors might be coupled to G-proteins that differ in their sensitivity to PTX treatment. PMID- 2550264 TI - Activation of type I cyclic AMP-dependent protein kinases is impaired by a point mutation in cyclic AMP binding sites. AB - The type I regulatory (R-I) subunit of cyclic AMP-dependent protein kinase (A kinase) was expressed in E. coli, and a single amino acid substitution in cyclic AMP binding sites A or B was introduced by site-directed mutagenesis. The cyclic AMP binding activity and cyclic AMP-stimulated phosphotransferase activity of the holoenzymes formed by wild-type or mutant R-Is and the purified bovine catalytic subunit of A-kinase were then examined. The wild-type holoenzyme was activated by low concentrations of cyclic AMP, a finding in accord with its high-affinity binding to cyclic AMP. In contrast, although the two mutant holoenzymes showed high-affinity cyclic AMP binding at their non-mutated sites, both holoenzymes were resistant to activation by cyclic AMP. Thus, binding of cyclic AMP to the non-mutated cyclic AMP binding site is not sufficient to dissociate the catalytic subunit from the mutant R-Is upon cyclic AMP binding. These results suggest that both A and B cyclic AMP binding sites are required for efficient coupling between cyclic AMP binding and activation of the enzyme. PMID- 2550265 TI - Characterization of the UV-induced electrophysiological response in smooth muscle. AB - The effect of ultraviolet (UV) light on taenia caeci smooth muscle of guinea-pig was studied. This stimulus induced a decrease in membrane conductance, hyperpolarization of the smooth muscle cells, a decrease in spike activity and a diminished level of cGMP and enhancement of lipid peroxidation. The hyperpolarization evoked by UV light was attenuated in the presence of sodium nitrite, reaching a maximum at 0.5 mM sodium nitrite. The level of cGMP was enhanced in the presence of nitrite and lipid peroxidation was not changed. Under these conditions lipid peroxidation was not changed during UV irradiation either. The UV-induced hyperpolarization was abolished in the absence of extracellular calcium, inhibited in the presence of 8-Br-cGMP and enhanced by an increase in the extracellular calcium concentration. These results are consistent with the supposition that UV-A irradiation blocks calcium influx via a cGMP-dependent mechanism. PMID- 2550266 TI - cAMP-dependent phosphorylation of soluble and crude microtubule fractions of rat cerebral cortex after prolonged desmethylimipramine treatment. AB - We have analyzed the cAMP-dependent phosphorylation system in the cerebral cortex and hippocampus of rats after acute and chronic administration of desmethylimipramine. Prolonged desmethylimipramine administration modified the cAMP-dependent endogenous phosphorylation of a protein band with apparent molecular weight 280 kDa from the cerebrocortical-soluble fraction. The effect appeared to be specific and associated with the chronic but not the acute administration of desmethylimipramine since we did not obtain any modification in other endogenous cAMP phosphoproteins of either the particulate or soluble fraction of the cerebral cortex. 280 kDa was identified as the soluble microtubule associated protein 2 on the basis of molecular weight, endogenous phosphorylation and immunological recognition. Prolonged desmethylimipramine administration did not induce any modification in the soluble cAMP-dependent endogenous phosphorylation of 280 kDa in other brain areas such as hippocampus, striatum or cerebellum, suggesting a region-specific effect of chronic desmethylimipramine treatment. Microtubule-associated protein 2 is a neuronal protein highly enriched in the dendritic portion of neurons and represents one of the major substrates in the cell for the type II cAMP protein kinase. Since the type II cAMP protein kinase that catalyzes the phosphorylation of microtubule associated protein 2 copurifies with microtubules, we performed endogenous phosphorylation using increasing concentrations of cAMP in a crude microtubule preparation where microtubule-associated protein 2 appeared to be more concentrated. Under our conditions the maximal effect occurred at 1 microM cAMP, revealing increased 32P incorporation in microtubule-associated protein 2 from a crude microtubule preparation obtained from the cerebral cortex of rats treated with desmethylimipramine. Photoaffinity labelling with 8-azido-[32P]cAMP of the various fractions obtained during the preparation of crude microtubules (S1, S2 and crude microtubules) revealed an increase in the labelling of a protein band with apparent molecular weight of 52 kDa after desmethylimipramine treatment. The labelling of a 47 kDa protein band, which is also present in S1 and S2 fractions was, however, not altered by drug treatment. In conclusion, our studies demonstrated that prolonged desmethylimipramine treatment elicited specific changes in the phosphorylation system associated with a crude microtubule fraction. PMID- 2550267 TI - Plasma membrane associated ATP as a regulator of the secretory activity of the pancreatic beta-cell. AB - beta-Cell-rich pancreatic islets from ob/ob-mice were used for evaluating how ATP associated with the plasma membrane participates in the regulation of insulin release. Increase of Ca2+ initiates insulin release from permeabilized beta-cells only in the presence of Mg-ATP. When bound to the inner part of the plasma membrane ATP depolarizes the beta-cells by closing a glucose-regulated K+ channel. It is possible that ATP in a plasma membrane compartment modulates insulin release also by stimulating ion pumps and exchange processes. ATP can regulate the secretory activity by binding also to the exterior of the beta cells. The addition of ATP resulted in stimulation of insulin release related to polyphosphoinositide breakdown. It is suggested that the granule fusion with the plasma membrane is followed by release of sufficient amounts of ATP and ADP for activating a P2-purinoceptor. This receptor may consequently be part of a system for amplifying the secretory response to glucose and other agents facilitating the entry of Ca2+. PMID- 2550268 TI - Studies on the chemical and immunological destruction of insulin-secreting islet cell tumours. AB - Chemical and immunological destruction of insulin-secreting islet cell tumours were evaluated in vivo and in vitro using the transplantable radiation-induced NEDH rat insulinoma and the derived clonal RINm5F cell line. Administration of a large amount of polyclonal insulin antibody did not affect the development of hypoglycaemia, tumour weights or the survival of insulinoma-bearing rats. Streptozotocin (100 mg/kg body weight) evoked a rapid and sustained decrease of insulin concentrations, accompanied by tumour regression and elevation of plasma glucose. Administration of alloxan (200 mg/kg) was without effect. In vitro, streptozotocin and alloxan exerted approximately equipotent time-dependent and concentration-dependent cytotoxic effects on cultured insulinoma cells as established by cell staining with trypan blue. The cytotoxic actions of both drugs were decreased by agents believed to scavenge free radicals or to act as inhibitors of poly(ADP-ribose)synthetase. Exposure of clonal RINm5F cells to the nitrosocompounds, N-methyl-N'-nitro-N-nitrosoguanidine and N-nitroso-N methylurea, resulted in a particularly marked reduction in cell viability compared with streptozotocin. PMID- 2550269 TI - Monoclonal antibodies as probes to the differentiated exocrine pancreas react to monoclonal islet tumor tissue. AB - Pancreatic exocrine and ductal reacting monoclonal antibodies were derived from a mouse immunized with fixed pluripotent rat islet tumor cells (MSL) and boosted in vitro with fixed and concentrated tumor cell culture supernatant. Antibodies were obtained against duct cells, intercalated ducts, acinar cells and zymogen granules as well as against parietal cells. Unexpectedly, no monoclonal antibodies were directed against the endocrine pancreas, whereas six out of seven exocrine reacting antibodies stained total or subpopulations of cells in sections of monoclonal hypoglycemic MSL-tumors. These data may support the hypothesis of a common endodermal origin of the exocrine and endocrine pancreas. PMID- 2550270 TI - Mechanisms of pancreatic beta-cell growth and regeneration: studies on rat insulinoma cells. AB - Information about the mechanism of beta-cell growth and regeneration may be obtained by studies of insulinoma cells. In the present study the growth and function of the rat insulinoma cell lines RINm5F and 5AH were evaluated by addition of serum, hormones, and growth factors. It was found that transferrin is the only obligatory factor whereas growth hormone, epidermal growth factor, fibroblast growth factor, and TRH had modulating effects. A heat-labile heparin binding serum factor which stimulated thymidine incorporation but not cell proliferation was demonstrated in human serum. Measurements of insulin mRNA content showed that the insulinoma cells only contained about 2% of that of normal rat beta-cells. These results are discussed in relation to the role of growth factors, oncogenes, and differentiation in the growth and regeneration of beta-cells. PMID- 2550271 TI - Radioreceptor assay for detergent-solubilized rat insulinoma growth hormone receptors. AB - Growth hormone receptors from a rat insulinoma cell line, RIN-5AH were solubilized in the non-ionic detergent Triton X-100. A radioreceptor assay based on polyethylene glycol precipitation of the growth hormone: receptor complex showed time-dependent and saturable hormone binding. The affinity in detergent solution for biosynthetic human growth hormone of approx. 6 ng/ml was found similar to that of intact RIN cells. The solubilization and receptor assay conditions described are useful for further characterization and purification of RIN cell growth hormone receptors, which might provide an initial insight into the molecular mechanism of the growth hormone effects on islet beta-cells. PMID- 2550272 TI - Reversal of glucose-induced inhibition of insulin release by dibutyryl cyclic AMP. AB - Insulin release in response to glucose was measured after culture of islets from ob/obmice in a medium deficient in Ca2+. When present at a concentration of 6 mmol/l, glucose inhibited, insulin release. After activation of the alpha2 adrenergic receptors by 1 mumol/l clonidine also 20 mmol/l glucose became inhibitory. The inhibitory action of glucose on insulin release disappeared in the presence of 1 mmol/l dibutyryl cyclic AMP. Raising the glucose concentration from 3 to 20 mmol/l resulted in increased cytoplasmic Ca2+ after an initial depression. Whereas clonidine removed the Ca2+ increase, this phase was partly restored after simultaneous addition of dibutyryl cyclic AMP. It is concluded that cyclic AMP plays a major role in controlling the balance between the stimulatory and inhibitory components in the glucose action on insulin release. PMID- 2550273 TI - Noradrenergic control of ascorbic acid and glutathione concentrations in brown fat of cold-exposed rats. AB - Ascorbic acid and glutathione concentrations increase in brown fat of cold exposed rats. This phenomenon can be reproduced by noradrenaline or isoproterenol administration, and thus seems to be under sympathetic control. Histological study shows ascorbic acid storage in brown adipocyte nuclei. PMID- 2550274 TI - An endogenous 'hypertensive factor' enhances the voltage-dependent calcium current. AB - The effects of an immunoaffinity-purified putative endogenous hypertensive factor (HF) on voltage-dependent calcium current in frog cardiac myocytes were assessed. In 9 out of 10 cells, HF reversibly increased the peak amplitude of the calcium current. HF increased peak calcium current density at -5 mV from a control level of 1.8 +/- 1.3 pA/pF (mean +/- SD) to 4.4 +/- 2.0 pA/pF. HF shifted the peak of the calcium current-voltage relationship in the hyperpolarizing direction. HF shifted the voltage dependence of the inactivation of the calcium current to more negative potentials with prepulses from -80 to 0 mV, but the inactivation was not affected with prepulses more positive than 0 mV. Modulation of the voltage dependent calcium current by HF may be the mechanism underlying its pressor effects. PMID- 2550276 TI - Chloride binding to photosystem II in the dark is in slow exchange. AB - We have studied the 35Cl- NMR line broadening in the presence of photosystem II (PS II) membranes from spinach in the dark. In the presence of NH3 (which other work has shown to competitively inhibit chloride binding to PS II) we observed no decrease in 35 Cl- linewidths. We conclude that binding of Cl- to the O2 evolving center in PS II in the dark (previously demonstrated by EPR) is in slow exchange on the NMR timescale. We assign the observed line broadening to interaction with non-specific binding sites and with free paramagnetics. PMID- 2550275 TI - A novel, specific binding protein assay for quantitation of intracellular inositol 1,3,4,5-tetrakisphosphate (InsP4) using a high-affinity InsP4 receptor from cerebellum. AB - A membrane preparation from porcine cerebellum displays high-affinity binding sites for [3H]inositol 1,3,4,5-tetrakisphosphate ([3H]InsP4) with a dissociation constant (Kd) of 1.0 nM and a density of 220 fmol/mg protein. Specific binding was maximal in the presence of 25 mM phosphate and at pH 5.0. The receptor site was specific for InsP4, since Ins(1,3,4,5,6)P5 and Ins(1,4,5,6)P4 displaced binding of InsP4 with EC50 values of 0.2 and 0.3 microM, respectively. Ins(1,4,5)P3 and other inositol phosphates were less effective. Using this InsP4 receptor, an assay for measuring tissue content of InsP4 was developed. The detection limit of the assay was 0.1 pmol. In the same tissue samples the amount of Ins(1,4,5)P3 was determined in parallel with a similar assay using a binding protein preparation from beef liver. PMID- 2550277 TI - Lipid phosphorylation in isolated rat liver nuclei. Synthesis of polyphosphoinositides at subnuclear level. AB - Isolated rat liver nuclei and subnuclear fractions synthesize polyphosphoinositides in vitro in a mode dependent on the presence of nuclear membrane, detergent and exogenous substrates. The nuclear membrane is not essential as a source of lipid kinases, since the addition of exogenous phosphatidylinositol or phosphatidylinositol monophosphate to reaction mixtures lacking membranes restores the synthesis of phosphatidylinositol mono- and bisphosphate, respectively. Inositide phosphorylation is best accomplished by high-salt extracted nuclei and pre-detergent lamina. These data suggest that the nucleus, and especially the nuclear periphery, is a cell compartment in which polyphosphoinositide synthesis occurs; this might be related to the progression of phosphatidylinositol metabolism-dependent signals to the genetic apparatus. PMID- 2550278 TI - High incorporation of [3H]inositol into phosphoinositides of human platelets during reversible electropermeabilisation. AB - A new method for high incorporation of [3H]inositol into human platelets is described. The method involves incorporation of [3H]inositol during reversible electropermeabilisation by high voltage discharge, followed by resealing the cells during incubation at 37 degrees C. Between 10- and 20-fold increase of isotope uptake is achieved compared to control intact cells. Permeabilised resealed platelets maintain good responses to thrombin and collagen. Analysis of the incorporation of the label amongst the phosphoinositides shows 70% to be in PI, 20% in PIP, and 10% in PIP2. Stimulation with thrombin and analysis of the formation of IP1, IP2 and IP3 shows the labelling to occur in a hormone-sensitive pool. These studies indicate that reversible electropermeabilisation can be used to achieve good uptake of non-membrane penetrating substances such as inositol. PMID- 2550279 TI - Synthesis of spin-labeled 2-azido-ATP: evidence for distinct nucleotide-binding sites in calcium pump protein from sarcoplasmic reticulum. AB - A spin-labeled and photoreactive derivative of ATP was synthesized with the spin label attached to the 2'- or 3'-position of the ribose moiety and an azido group to C2 of the adenine ring (SL-2N3-ATP). Irradiation of this compound at 350 nm generates a nitrene, which then reacts with nucleophiles in its vicinty. SL-2N3 ATP, in the presence of Ca2+, was hydrolyzed by the calcium pump protein (Ca2+ ATPase) of fast twitch skeletal muscle sarcoplasmic reticulum. The SL-2N3-ATP enzyme complex in the absence of Ca2+ exhibited strongly immobilized ESR spectra. ESR spectra obtained after covalent incorporation of SL-2N3-ATP into Ca2+-ATPase and removal of freely tumbling SL-2N3-ATP exhibited motionally constrained species indicative of distinct and possibly adjacent ATP-binding sites. By contrast, with SL-ATP devoid of the azido group or with the corresponding 'non cleavable' beta, gamma-methylene triphosphate analogue (SL-AMP-PCP), two distinct sites were not as well resolved in the ESR spectra due to spectral overlap with the signal from the freely tumbling fraction even with the enhanced spectral resolution provided by perdeuteration of the spin label. Thus, SL-2N3-ATP may have general application for ESR studies of ATP-dependent proteins under conditions in which non-covalent interactions are too weak for motionally restricted species to be resolved. PMID- 2550280 TI - Mapping of beta-adrenoceptor coupling domains to Gs-protein by site-specific synthetic peptides. AB - Peptides corresponding to the known sequence of turkey erythrocyte beta 1 adrenergic receptor were synthesized and the effects on receptor-mediated cyclase activation were measured. Peptides corresponding to the first and second intracellular loops (T61-71 and T138-159) inhibited at micromolar concentrations the hormone-dependent cyclase activation in turkey erythrocyte membranes. In contrast, the peptide corresponding to the C-terminal part of the third intracellular loop (T284-295) increased the cyclase activity in a hormone independent manner. Peptides T338-353 and T2-10 and a number of synthetic peptides unrelated to the beta-adrenoceptor had no effect. PMID- 2550281 TI - Interactions of formylmethionyl-leucyl-phenylalanine, adenosine, and phosphodiesterase inhibitors in human monocytes. Effects on superoxide release, inositol phosphates and cAMP. AB - Cessation of the fMLF-induced burst of human monocyte superoxide release was associated with a rise in cAMP. This was not due to inhibition of phosphodiesterase (PDE), the major form of which was the PDE IV isozyme. The action of burst inhibitors did not correlate with cAMP levels: Rolipram, a PDe IV inhibitor, increased cAMP 6-fold, with minimal effects on the burst; whereas theophylline increased cAMP less than 2-fold but decreased the burst to less than half. Although theophylline and the adenylate cyclase activator, adenosine, inhibited fMLF-induced superoxide release, they did not inhibit production of inositol phosphates. Thus, these studies on inhibition of superoxide release implicated neither cAMP nor inositol phosphates. PMID- 2550282 TI - Comparison of motility stimulants for cryopreserved human semen. AB - Caffeine, pentoxifylline, 2-deoxyadenosine, cyclic adenosine monophosphate (cAMP), relaxin, adenosine, kallikrein, and calcium were compared for their ability to stimulate motility of cryopreserved sperm. Caffeine, pentoxifylline, and 2-deoxyadenosine significantly increased the percentage of motile sperm at 15, 30, 45, and 60 minutes after administration. Sperm velocity was significantly increased by caffeine at 0, 15, 30, and 45 minutes, and by pentoxifylline at 0, 45, and 60 minutes. Consistent stimulation was not observed for other chemicals. Caffeine, pentoxifylline, and 2-deoxyadenosine were then examined for their ability to provide motility stimulation after removal with washing. With the exception of caffeine, percent motility and velocity for stimulated and untreated sperm were similar after washing. A significant reduction in motility was observed at 48 hours after washing for caffeine. The percentage of hamster oocytes penetrated at 24 hours after washing was significantly reduced for caffeine, 2-deoxyadenosine, and pentoxifylline combined with 2-deoxyadenosine. Pentoxifylline-treated sperm showed no reduction in fertilizing capacity. These results indicate that, of the chemicals examined, pentoxifylline is superior for motility stimulation of cryopreserved sperm. PMID- 2550283 TI - Possible role of H2O2-mediated complement activation and cytokines-mediated fibroblasts superoxide generation on skin inflammation. AB - The degree of complement activation produced by hydrogen peroxide was estimated by the inhibition of serum homolytic activity (% IHA). Sera from patients with systemic lupus erythematosus and psoriasis vulgaris were resistant to hydrogen peroxide-mediated complement activation. %IHA negatively correlated with ceruloplasmin level or catalase activity in systemic lupus erythematosus sera, but did not correlate with transferrin level. The addition of free metal ions, FeCl2 or CuCl2, promoted hydrogen-peroxide-mediated complement activation. These results suggest that hydroxyl radical is involved in complement activation and that the factors responsible for the insensitivity of pathological sera to H2O2 are catalase and ceruloplasmin in the sera. Human skin fibroblasts generate superoxide and tumor necrosis factor enhanced it, but interleukin-1 beta inhibited it. Normal serum cultured with fibroblasts for 24 h showed complement activation via catalase-inhibitable process, suggesting that hydrogen peroxide has an important role in fibroblast-mediated complement activation. It is speculated that fibroblasts and complement activation by oxygen radicals have an important role in inflammation and subsequent tissue damage at the site of skin lesion. PMID- 2550284 TI - Release of neutrophil and eosinophil chemotactic factor from sensitized skin in vitro cutaneous anaphylactic reactions. PMID- 2550285 TI - Enhanced bactericidal activity of macrophages by exogenous leukotriene B4. PMID- 2550286 TI - Effect of metallothionein on granulocyte chemotaxis and superoxide generation. PMID- 2550287 TI - Mechanisms for hyperpigmentation in postinflammatory pigmentation, urticaria pigmentosa and sunburn. AB - Our in vitro studies demonstrate that normal human epidermal melanocytes become swollen and more dendritic with an increase in amount of immunoreactive tyrosinase when they are cultured for several days with arachidonic acid metabolites, vitamin D3 or histamine. From these data we propose the following possible mechanisms for hyperpigmentations noted at postinflammatory sites and suntanned areas as well as at skin lesions of urticaria pigmentosa. Arachidonic acid metabolites and histamine, which are found in increased amounts in inflammatory skin, are thought to play a key role in the induction of postinflammatory hyperpigmentation. In sunburnt skin the increased proinflammatory mediators, particularly arachidonic acid metabolites, are also thought to stimulate melanocytes in the production of hyperpigmentation. Thus tanning after sun exposure may be induced not only by the effect of vitamin D3 and direct UV irradiation on the melanocytes but also by the effect of various arachidonic acid metabolites which are increased in sunburnt skin. Mast cells massively proliferate in the skin lesions of urticaria pigmentosa. Thus hyperpigmentation in the skin lesions of urticaria pigmentosa is quite likely to be induced by the chemical mediators, including histamine and leukotrienes, that are released from these cells. PMID- 2550288 TI - From eosinophil chemotactic factor of anaphylaxis to leukotriene B4--chemistry, biology and functional significance of eosinophil chemotactic leukotrienes in dermatology. AB - The present paper examines evidence for the identity of low molecular weight eosinophil chemotactic factor (ECF) and of leukotriene B4 (LTB4) and its metabolites. Total congruity between the two entities is found regarding (1) cells of origin; (2) conditions for in vitro generation and pharmacological modulation; (3) physiochemical properties; (4) in vitro chemotaxis towards human monocytes, fibroblasts and guinea pig eosinophils; (5) in vivo activities in humans, and (6) occurrence of the factors in various dermatological diseases. Quantitative differences were observed only for in vitro neutrophil migration which may be due to neutrophil chemotactic mono-HETEs and possibly platelet activating factor in the ECF preparations. The name ECF should therefore be replaced by LTB4 and its metabolites, as has happened for SRS which is now called LTC4/D4. PMID- 2550289 TI - Modulation of plasma exudation by PGE2 and that of leukocyte migration by LTB4 in inflammatory models. AB - Des-Phe8-Arg9-BK could be detected in the entire course of rat carrageenin pleurisy up to 24 h, together with a reduction of the residual levels of high molecular weight kininogen and prekallikrein. On the basis of this continuous release of bradikinin, prostaglandin E2 was released up to 5 h in the pleural exudate and enhanced the plasma leakage. In rat cardiac infarction, the initial increase in the number of polymorphonuclear leukocytes in the cardiac tissue was accompanied by leukotriene B4 in the tissue and this was followed by the second increase and activation of the complement system. PMID- 2550290 TI - Role of bradykinin in exudative reactions of inflammation in rats. AB - This study was done with the aid of experimental models of inflammation provoked in air pouch, which was made subcutaneously on the back of rats, by applying a carrageenin solution or a suspension of kaolin in a carboxymethylcellulose solution as an irritant. Bradykinin in the inflammatory sites, determined by an enzyme immunoassay method, were comparatively high in the earliest stage of inflammation in both the models and responsible for the exudative reactions. With the passage of time, i.e. within 20 min in the case of kaolin and 120 min in carrageenin, tissue kininase entered the inflammatory site and reduced bradykinin to a very low level, though bradykinin was still being generated very actively. In this later stage, bradykinin collaborates with prostaglandin to bring about a significant increase in the vascular permeability. PMID- 2550291 TI - Real-time echo-Doppler Duplex Scanner in the evaluation of patients with Poland sequence. AB - In two neonates with partial Poland sequence, an infant with complete Poland sequence and his mother subclavian artery diameter and its flow velocity waveforms were determined by real-time echo-Doppler Duplex Scanner. These measurements were significantly lower than on the normal side in the same patient. Subclavian hypoplasia appears as a local vascular defect which, in addition to thoracic muscle defects and bone defects of the hand, may be one of the main components of the Poland sequence. PMID- 2550292 TI - Time of separation of the umbilical cord: a comparative study of treatment with alcohol and Rikospray. AB - The time of cord separation and the umbilical changes post cord falling were studied in 394 infants, 35 to 42 weeks gestation, 1900 to 4500 g birth weight. Two methods of treatment; alcohol 70% with 0.5% chlorhexidine, and Rikospray were compared. The mean +/- SD age of cord separation was 6.36 +/- 2.64 days (range 3 20 days) in all infants studied. The time of cord separation and post fall umbilical complications were similar in the two treated groups. The only point of difference was the repulsive odor in the alcohol-treated group, making handling of the infants very unpleasant. PMID- 2550293 TI - Expression of the plasma membrane proteolipid in mouse neuroblastoma cells: transient increase in synthesis during differentiation with N6,O2-dibutyryl adenosine 3',5'-cyclic monophosphate. AB - We have examined the regulation of plasma membrane proteolipid (PM-PLP) synthesis and steady-state levels in mouse NB2a/d1 neuroblastoma cells during differentiation with dibutyryl cyclic AMP (dbcAMP) and retinoic acid (RA), agents which have been previously shown to induce the elaboration of exclusively axonal or dendritic neurites, respectively. We report that a PM-PLP-immunoreactive species is expressed by this neuroblastoma cell line, and that its expression is regulated by specific states of differentiation. Differentiation of cells with dbcAMP was accompanied by an initial 2-fold increase in this PM-PLP immunoreactive species at 24 h after treatment, which returned to control levels by 96 h after treatment. By contrast, no significant increase in synthesis was detected when cells were treated with RA. Protein blot analysis of PM-PLP in dbcAMP-treated cells indicated that there was little change in its steady-state level until 96 h following treatment, at which time a reduction of 40% was observed. Throughout induced differentiation with dbcAMP, NB2a/d1 cells continued to express a PM-PLP-immunoreactive species which comigrated on immunoblot analysis with PM-PLP form characteristic of embryonic brain (14-16 kDa), and apparently did not express the PM-PLP form characteristic of adult brain (18 kDa). PMID- 2550294 TI - A rat seminiferous epithelial factor that inhibits Leydig cell cAMP and testosterone production: mechanism of action, stage-specific secretion, and partial characterization. AB - To elucidate further the seminiferous tubule-Leydig cell interaction we studied the effect of spent medium from 20 h rat seminiferous tubule cultures (STCM) on cyclic adenosine monophosphate (cAMP) and testosterone (T) production of Percoll purified Leydig cells. 8% of STCM inhibited the human chorionic gonadotropin (hCG)-stimulated cAMP production by 30% in a 3 h, and 33% of STCm by 60% in a 20 h incubation. Likewise, a 40% decrease was found in the presence of 8% of STCM in the hCG-stimulated T production in a 3 h incubation. A similar inhibitory activity could be demonstrated in steroid-free rat interstitial fluid. STCM did not affect the viability of the Leydig cells (90-95% after a 20 h incubation) as judged by trypan blue exclusion and the adenosine triphosphate (ATP) content of the cells. Heating (80 degrees C for 10 min) abolished the inhibiting activity, and fractionation with Millipore Ultrafree filters showed that the inhibiting factor had an Mr of 30,000-100,000. When media from different stages of the seminiferous epithelial cycle were analyzed, only stages IX-I showed inhibition of T production (P less than 0.05). The cAMP production inhibiting activity was present in all stages, but stages IX-I showed significantly (P less than 0.05) greater inhibition than stages II-VI. STCM (16%) also inhibited cholera toxin- and forskolin-stimulated cAMP formation (approximately 50 and 60%, respectively; P less than 0.01), and the inhibitory activity persisted after a 3 h preincubation of Leydig cells with 100 micrograms/l pertussis toxin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550295 TI - Comparison of insulin-like growth factor 1 and insulin effects on prolactin induced lactogenesis in the rabbit mammary gland in vitro. AB - In organ culture of pregnant rabbit mammary gland, a low casein synthesis occurs with prolactin (PRL) alone. Insulin markedly potentiates the effect of PRL. Only pharmacological concentrations of insulin (5 micrograms/ml) exert the maximal enhancement, suggesting a possible interaction with the insulin-like growth factor 1 (IGF1) receptor. The presence of IGF1 and insulin binding sites was analyzed and the biological effects of both peptides were compared. Binding of iodinated human IGF1 or porcine insulin to mammary microsomes prepared from mid pregnant rabbits revealed distinct high affinity binding sites for both peptides (Kd approximately 2 nM). In rabbit mammary explants, we confirmed that only non physiological concentrations of insulin (greater than or equal to 100 ng/ml) exerted a significant stimulation of the PRL effect. Surprisingly, IGF1 was not found to be more potent than insulin on a molar basis, which did not provide evidence for the exclusive involvement of the IGF1 receptor. Near-physiological concentrations of IGF1 (approximately 100 ng/ml), however, exerted a significant enhancement which suggested a possible action for IGF1 on PRL-induced lactogenesis in vivo. PMID- 2550296 TI - Transmembrane signals mediating adrenocorticotropin release from mouse anterior pituitary cells. AB - The effect of arginine vasopressin (AVP) and corticotropin releasing factor (CRF) an adrenocorticotropin (ACTH) secretion, phosphatidylinositol breakdown and cAMP accumulation was examined in primary cultures of mouse anterior pituitary cells. AVP and CRF added alone stimulated ACTH secretion in a dose-dependent manner. At 10(-8) M concentration of peptide, AVP and CRF stimulated ACTH secretion 2.8- and 4.6-fold, respectively. AVP and CRF added in combination at equal doses gave an additive effect. CRF enhanced cAMP accumulation, but AVP had no effect on basal or CRF-induced cAMP accumulation. Both forskolin (10(-5) M) and 8-bromo-cAMP (10( 3) M) increased ACTH secretion in these cells by 2.8- and 1.7-fold, respectively. AVP induced the breakdown of phosphoinositides, and CRF alone, or in combination with AVP did not modify this effect. Phorbol 12-myristate 13-acetate (10(-7) M), dioctanoylglycerol (10(-4) M) and phospholipase C (100 mU/ml) also stimulated ACTH secretion in these cells by 4.2-, 2.4-, and 3.7-fold, respectively. Depletion of intracellular and extracellular Ca2+ decreased ACTH secretion, but had no significant effect on CRF-induced cAMP accumulation. However, AVP-induced phosphoinositide breakdown was dependent on extracellular Ca2+. These results indicate that CRF stimulates ACTH secretion via the cAMP-dependent pathway and AVP via the phosphoinositide breakdown-phospholipase C pathway. In the presence of AVP and CRF, both pathways appear to operate independently to produce an additive effect on ACTH secretion. PMID- 2550297 TI - Biosynthesis of polyunsaturated fatty acids of (n-6) and (n-3) series in isolated adrenocortical cells of rats. Effect of ACTH. AB - Both the capacity of isolated adrenocortical cells to incorporate and transform [1-14C]linoleic and [1-14C]alpha-linolenic acids and the effect of ACTH on the biosynthesis of polyunsaturated fatty acids from [1-14C]alpha-linolenic acid were investigated. The cells were able to incorporate both labeled precursor acids and convert them into higher homologs. This transformation increases along the incubation time tested. When linoleic acid was the precursor, the biosynthesis of higher homologs was carried out following the desaturating-elongating route. Both pathways, the desaturating-elongating and the elongating, were detected when the substrate was alpha-linolenic acid. The results proved the existence of delta 6, delta 5 and delta 4-desaturases in this type of cells. Isolated adrenocortical cells obtained from rats treated with ACTH showed an increase in the amount of [1 14C]alpha-18:3 that remained in the cells without metabolization and, consequently, a decrease in the last product formed (20:5 n-3) was evident compared to the controls. Simultaneously, the desaturation-elongation products decreased significantly. Similar results were obtained when cells isolated from untreated rats were incubated for 3 h in the presence of ACTH. In this case, the values obtained returned to normal levels 6 h after incubation. These results were mimicked by dibutyryl-cyclic AMP. It can be concluded that the effect of ACTH on the biosynthesis of polyunsaturated fatty acids from alpha-linolenic acid was mediated through an enhancement of the intracellular levels of cAMP. PMID- 2550298 TI - Effects of human chorionic gonadotropin, prostaglandin F2 alpha and protein kinase C activators on the cyclic AMP and inositol phosphate second messenger systems in cultured human granulosa-luteal cells. AB - The effects of human chorionic gonadotropin (hCG) and prostaglandin F2 alpha (PGF2 alpha) on the adenylate cyclase-cAMP and inositol phospholipid phospholipase C-inositol trisphosphate and diacylglycerol transmembrane signalling systems were evaluated in cultured human granulosa-luteal cells. Granulosa-luteal cells obtained from patients undergoing in vitro fertilization were cultured for 72 h prior to addition of hormones. During the last 24 h of culture granulosa-luteal cells were incubated with [3H]inositol. Neither hCG nor gonadotropin-releasing hormone (GnRH) stimulated the inositol phospholipid phospholipase C signalling system. PGF2 alpha stimulated increases in inositol mono-, bis-, and trisphosphate accumulation in 30 min incubations. NaF (20 mM) mimicked the stimulatory effect of PGF2 alpha on inositol phosphate accumulation suggesting the involvement of a guanine nucleotide regulatory protein in the activation of phospholipase C. In contrast, hCG but not PGF2 alpha or NaF stimulated cAMP accumulation in 30 min incubations. Simultaneous treatment with hCG and PGF2 alpha did not alter the stimulatory effect of PGF2 alpha on inositol phosphate accumulation but reduced (37%) the stimulatory effect of hCG on cAMP accumulation. The protein kinase C activator, 12-O-tetradecanoylphorbol 13 acetate (TPA) inhibited the stimulatory effects of hCG (76%) and PGF2 alpha (62%) on cAMP and inositol phosphate accumulation, respectively. Thus, cultures of human granulosa-luteal cells possess multiple transmembrane signalling systems which may be modulated by the activation of protein kinase C. PMID- 2550299 TI - Effect of ethane dimethane sulphonate on proopiomelanocortin (POMC) mRNA and POMC derived peptides in the rat testis. AB - Recent studies have shown that both proopiomelanocortin (POMC)-derived peptides and a range of POMC gene transcripts are present in the testis. Previous immunocytochemical studies have reported immunoreactive (ir)-beta-endorphin (EP) and ir-ACTH to be localized in the Leydig cells, and ir-NacEP in spermatogonia and primary spermatocytes. In the present study, we have further examined the hypothesis that testicular Leydig cells are the principal site of synthesis of these peptides, by determining the effects of the administration of the cytotoxic drug ethane dimethane sulphonate (EDS) which selectively destroys the Leydig cells of the testis. As expected, serum testosterone levels fell and serum FSH/LH levels increased within 3 days of EDS administration, returning to normal levels 4-8 weeks later. In contrast, the testicular content of POMC-derived peptides and POMC mRNA levels in these animals was not significantly altered throughout the experimental period. In addition, POMC mRNA was not detected in a purified Leydig cell preparation derived from adult male rats, and POMC-derived peptides were also undetectable in the media of a similar preparation following cell culture. These data suggest that in the adult the predominant site of rat POMC gene expression is in testicular interstitial cells other than Leydig cells. PMID- 2550300 TI - Calf serum modifies the mitogenic activity of epidermal growth factor in WRT thyroid cells. AB - It has already been shown that Wistar rat thyroid (WRT) cells in low concentrations of calf serum (0.5%) are under the influence of both thyrotropin (TSH) and insulin as regards growth. The present data show that epidermal growth factor (EGF), in concentrations up to 10 micrograms/ml, is not able to modify DNA synthesis in WRT cells. On the other hand, insulin-like growth factor I (IGF-I) stimulates DNA synthesis from a dose which is 10-fold lower than that of insulin alone. Combined stimulation of EGF and TSH in WRT cells is equal to that of TSH alone in relation to DNA synthesis, while the combined presence of TSH and IGF-I, or TSH and insulin, in the same medium results in an effect which is greatly superior to the theoretical sum of activities. Repetition of the same experiments using the original clone of WRT cells, but in high concentrations of calf serum (5%), shows that EGF stimulates DNA synthesis in a dose-dependent way from 0.1 to 100 ng/ml. Under these conditions, combined stimulation of EGF with TSH shows that DNA synthesis is equal to the predicted theoretical sum. No other differences in WRT cell sensitivity to either IGF-I or insulin, or IGF-I and TSH and insulin and TSH, can be noted. This finding is confirmed by the demonstration of specific and sensitive binding sites for EGF on WRT cells cultured in 5% calf serum; these binding sites are not present on WRT cells adapted to grow in 0.5% calf serum. Present data support the hypothesis that EGF and serum growth actions are mediated through the same analogous pathway, which is, however, different from those of TSH and/or IGF-I and/or insulin. PMID- 2550301 TI - Changes in the number, function, and regulation of nicotinic acetylcholine receptors during neuronal development. AB - Acetylcholine receptor (AChR) development was examined in neurons freshly isolated from chick ciliary ganglia. Between Embryonic Day 8 (E8) and 16, both the ACh response per unit membrane and the density of surface AChRs increased, while the apparent affinity for ACh decreased. AChRs had single-channel conductances of 25 and 40 pS. The distribution of single-channel events shifted during development; at E8 events of both conductances were equally rare, while by E14 there were many events and most were 40 pS. The open durations of 25 and 40 pS events had two mean values. The open lifetimes of the 25 pS events did not change between E8 and E14, while the lifetimes of the 40 pS events increased, and by E14 most were long. The ACh response of the neurons also became sensitive to regulation by a cAMP-dependent mechanism at about E10. The observed changes may reflect developmental control over processing required for receptor regulation and differential expression of AChR subtypes. PMID- 2550303 TI - Yeast virology. AB - The three families of double-stranded RNA (dsRNA) viruses and two families of retroviruses (retrotransposons) of the yeast Saccharomyces cerevisiae are all transmitted between cells only by cell fusion, probably reflecting the high frequency of mating of yeast cells in nature. One dsRNA virus and two retroviruses apparently use ribosomal "frameshifting" to produce major coat protein-polymerase fusion proteins. This mechanism allows regulation of the relative amounts of major coat protein and fusion protein that are made and avoids the possibility of mutant virus genomes being generated by splicing. Moreover, the fusion protein structure suggests a possible mechanism of genome packaging. The recent development of in vitro replication, transcription, and integration systems for these viruses, and the ease with which classical genetic and molecular studies are executed in yeast, are yielding detailed information about the roles of cellular and viral components in the viral replication cycles and the host defensive response. A host defense system against yeast dsRNA viruses is known and there is evidence to suggest a system active against the retroviruses. PMID- 2550302 TI - Induction of murine 8-cell blastomere polarity by F9 embryonal carcinoma cells. AB - The individual blastomeres of the preimplantation mouse embryo become polarized during the 8-cell stage of development. This polarity forms as a result of a specific cell-cell interaction that has been termed induction. The ability of embryonal carcinoma (EC) cells to induce 8-cell blastomere polarization has been investigated by aggregating nonpolar 8-cell blastomeres with various types of EC cells. F9, a nullipotent stem cell, induced polarization of a nonpolar 8-cell companion in 80% of the aggregates. Stimulation of differentiation of F9 cells with retinoic acid (RA), with or without dibutyryl cAMP, caused a reduction in the polarity-inducing ability of these cells. Other EC cells, PSA-1, NULLI-SCC1, 3TDM, C3HNE, and P10, all displayed less polarity-inducing activity than F9. In addition, it was observed that when any of these cell types assumed a more differentiated phenotype, either spontaneously or in response to specific stimuli, they displayed a decrease in their ability to induce 8-cell polarization. As a control, the inducing ability of cells from normal mouse tissues was examined. It was found that neither STO mouse fibroblasts nor primary cultures of mouse lymphocytes were able to induce significant polarization of 8 cell stage blastomeres. These data support the hypothesis that while undifferentiated stem cell populations retain the ability to induce 8-cell blastomere polarization, it is apparently lost upon cellular differentiation. PMID- 2550305 TI - Myocardial involvement due to a disseminated human cytomegalovirus infection in a heart transplant recipient. A case report. AB - A cytomegalovirus seronegative 9 year-old child, affected by terminal dilated cardiomyopathy, underwent cardiac transplantation and received the heart of a cytomegalovirus seropositive donor. After the positive outcome of an acute moderate rejection episode which was treated with steroids, the patient developed an infectious clinical syndrome with cytomegalovirus seroconversion and virus isolation from peripheral blood leukocytes. The 5th endomyocardial biopsy showed a typical cytomegalovirus-like nuclear inclusion in a single cell. In spite of specific treatment with (9-(2-hydroxy-1-(hydroxymethyl)ethoxy)methyl-guanine) (BW B759U) (Wellcome), the patient died 45 days after the operation from cytomegalovirus-induced interstitial pneumonia and virus-associated hemophagocytic syndrome. PMID- 2550304 TI - Antiinflammatory activity of a COOH-terminal fragment of the neuropeptide alpha MSH. AB - The endogenous neuropeptide alpha-melanocyte stimulating hormone (alpha-MSH 1 13), previously found to have marked antipyretic activity, inhibits histamine induced increases in vasopermeability. The primary antipyretic amino acid message sequence is believed to be the COOH-terminal trieptide, lysine-proline-valine. In recent preliminary research this tripeptide inhibited increases in vasopermeability, raising the possibility that this portion of the alpha-MSH molecule has general antiinflammatory activity. To test this idea, the effects of graded doses of alpha-MSH [11-13] on ear swelling induced by picryl chloride in mice were compared with the effects of saline and a large dose of corticosteroid. Alpha-MSH [11-13] inhibited swelling in a dose-related fashion. This result, together with previous findings, suggests that endogenous circulating alpha-MSH and its COOH-terminal fragments may contribute to modulation of physiological responses in host defense. If this is true, it may be possible to develop new peptide drugs or mimetics based on the tripeptide that are useful in treating inflammation. PMID- 2550306 TI - Corrosive gastritis mimicking linitis plastica carcinoma. AB - A 59-year-old female with depressive tendencies was admitted suffering from hematemesis and abdominal pain, two hours after ingestion of an unknown amount of toilet bowl cleaner (hydrochloric acid, pH 1.0). A barium study 24 days after ingestion revealed rigid narrowing and granulation of the entire stomach. The esophagus and duodenum were normal. The radiographic results were similar to those obtained for linitis plastica carcinoma of the stomach, but biopsy specimens of the stomach revealed no cancer cells. A total gastrectomy was performed about two months after ingestion to relieve the persistent feeling of nausea. Specimens revealed a rigid and thickening lining and a denuded mucosal surface of the stomach. The cut surface of the specimen showed a remarkable fibrous thickening of the submucosal layer. Microscopic examination failed to reveal a normal mucosal layer except in a narrow area of the fornix, and remarkable fibrosis of the submucosal lining was noted. No cancer cells were found. Corrosive gastritis has a linitis plastica appearance with a predilection for the antrum. Radiological examination revealed the very rare manifestation of a rigid narrowing of the whole stomach mimicking linitis plastica type cancer. PMID- 2550307 TI - Hepatitis B virus infection in liver cirrhosis and hepatocellular carcinoma in Jakarta Indonesia. AB - The relationship between hepatitis B virus (HBV) infection and 144 cases with liver cirrhosis (LC) and 82 cases with hepatocellular carcinoma (HCC), admitted to the Department of Internal Medicine, Dr. Cipto Mangunkusmo Hospital in Jakarta, was investigated within the period from January 1 until December 31, 1984. The prevalence of HBsAg in 144 cases with LC was 41.1%, anti-HBs 33.3% and anti-HBc alone 16.0%. In only 9.0% no markers were found. The prevalence of HBeAg in 60 cases HBsAg positive LC was 35.0%, anti-HBe was 41.7% and in 23.3% both markers were negative. Among 82 cases with HCC, the prevalence of HBsAg was 47.6%, anti-HBs was 25.6% and anti-HBc alone was 15.9%. The prevalence of HBeAg in 39 cases with HCC was 25.6%, anti-HBe was 51.3% and in 23.1% both markers were negative. Age distribution between HBsAg positive and negative in LC was observed significant difference (P less than 0.05). Mean age of HBsAg positive group in LC was 5 years younger than that of the HBsAg negative group. PMID- 2550308 TI - Inhibition of hydrogen-potassium-stimulated adenosine triphosphatase: effects on acid secretion, plasma gastrin, and the gastric mucosa. PMID- 2550309 TI - Cell-free synthesis of rat and rabbit gastric proton pump. AB - Ribonucleic acid was isolated from the fundic gastric mucosae of rats and rabbits by cesium chloride centrifugation of guanidine isothiocyanate-denatured mucosal homogenates, and poly A+ RNA was recovered from the pellets by oligodeoxythymidine column selection. When added to rabbit reticulocyte lysates, this poly A+ RNA stimulated [35S]methionine incorporation into trichloroacetic acid-precipitable material. Fluorographic analysis of the lysates showed protein synthesis to be dominated by polypeptides with molecular weights from 40,000 to 50,000, presumably prepepsinogen isoforms. Immune precipitation of the lysates with monoclonal antibodies directed against the gastric H+,K+-adenosine triphosphatase yielded bands at 94 kilodaltons and more diffuse banding at 180 kilodaltons. Further purification of the poly A+ RNA on sucrose gradients eliminated prepepsinogen messenger RNA; nascent H+,K+-adenosine triphosphatase synthesized by purified messenger RNA consisted of polypeptides with molecular weights between 88,000 and 94,000. The study indicates that cell-free translation of gastric mucosal messenger RNA may provide a useful model for analysis of gastric H+,K+-adenosine triphosphatase biosynthesis and processing. PMID- 2550310 TI - Na+/H+ exchange in isolated hamster enterocytes. Its major role in intracellular pH regulation. AB - The intracellular pH (pHi) of isolated hamster intestinal cells was determined with three techniques: the null-point method with digitonin, the distribution of the weak acid 5,5-dimethyloxazolidine-2,4-dione, and the trapped fluorescent indicator 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein. In physiologic saline, the pHi determined with the fluorescent dye 2',7'-bis(carboxyethyl)-5(6) carboxyfluorescein was 7.00 +/- 0.03 (n = 20) but was 6.68 +/- 0.04 (n = 25) when there was no Na+ in the medium, indicating an important role of external Na+ in maintaining pHi at neutral level. When Na+ was added to an incubation medium lacking the ion, the pHi increased. The time-course of this alkalinization depended on the Na+ concentration, whereas K+ had only a slight effect. Amiloride (1 mM) completely inhibited the Na+ effect, thereby showing the basic role of the Na+/H+ antiport in the regulation of pHi. The effect of 4-acetamido-4' isothiocyanostilbene-2,2'-disulfonic acid and furosemide showed that there also exists an anionic component in the process, probably of the Cl-/OH- (HCO3-) exchange type. These results show that the amiloride-sensitive Na+/H+ antiport plays an important but not exclusive role in maintaining the pHi neutral or slightly alkaline in isolated intestinal cells from the hamster. PMID- 2550311 TI - Treatment of hepatocellular carcinoma by transcatheter arterial embolization combined with intraarterial infusion of a mixture of cisplatin and ethiodized oil. AB - The therapeutic effectiveness of transcatheter arterial embolization (TAE) with intraarterial infusion of cisplatin/ethiodized oil mixture in treatment of resectable and unresectable hepatocellular carcinoma was compared with TAE with intraarterial infusion of doxorubicin mixed with and without ethiodized oil. The series included 97 patients with unresectable hepatocellular carcinoma and 40 patients with resectable hepatocellular carcinoma. With TAE using doxorubicin infusion, a partial response of the tumor was seen in only 11%, and the 2-yr survival was calculated to be only 5%. Histologic examination of the specimens obtained by hepatectomy also showed that this treatment was relatively ineffective in daughter tumor and portal tumor thrombi. In contrast, TAE with infusion of cisplatin/ethiodized oil mixture significantly increased the rate of partial response (38%), and significantly prolonged the 2-yr survival (45%). Histologically this treatment gave severe necrosis in daughter tumors (69%) and tumor thrombi (78%) as well as main tumor (75%). This treatment was significantly better than TAE with doxorubicin and ethiodized oil infusion in terms of the tumor regression and histologic responses of main tumor and portal vein tumor thrombi, but not in terms of the 2-yr survival. However, 2 patients (8%) died within 4 wk of the latter treatment, whereas no deaths were reported after the former treatment. Therefore, TAE combined with intraarterial infusion of cisplatin/ethiodized oil mixture may be a safe and useful treatment modality for hepatocellular carcinoma. PMID- 2550312 TI - [Malignant mixed Mullerian tumors of the uterus. Clinical, morphologic and immunohistochemical findings]. AB - Clinical and morphological findings in four patients with mixed mullerian tumors of the uterus showed, that these highly aggressive neoplasms are derived from the undifferentiated mullerian epithelium. Immunohistochemically, each component exhibits characteristic differentiation markers. The presence of cytokeratins or desmin signals an epithelial viz. rhabdomyosarcomatous differentiation, whilst the detection of vimentin in the absence of other markers demonstrates an undifferentiated mesenchymal component. This component appears to be responsible for the aggressive biological behavior of mixed mullerian tumors. PMID- 2550313 TI - Experimental aspects of isolation of myocytes from adult guinea-pig hearts. AB - Ionic channels can now be effectively studied on enzymatically isolated cardiac myocytes by means of the patch clamp technique. Three procedures reported to give consistently high yields of Ca-tolerant myocytes were tested for applicability to calcium channel studies under our laboratory conditions. None of them was found to be suitable for direct use. Therefore, a modified method for isolation of myocytes from adult guinea-pig hearts was developed. Calcium channel currents measured in Ca-tolerant myocytes isolated by this procedure have been presented and problems of myocytes isolation and of patch-clamp measurement discussed. PMID- 2550314 TI - A spin label study of perturbation effects of N-(1-methyldodecyl)-N, N, N trimethylammonium bromide and N-(1-methyldodecyl)-N, N-dimethylamine oxide on model membranes prepared from Escherichia coli-isolated lipids. AB - Interaction of bactericidal surfactants N-(1-methyldodecyl)-N, N, N trimethylammonium bromide (2-ATDBr) and N-(1-methyldodecyl)-N, N-dimethylamine oxide (2-ATDNO) with phospholipid membranes prepared from Escherichia coli -- isolated lipids was studied by ESR spectroscopy using m-doxyl stearic acid (m DSA, m = 5, 12, 16) and N-cetyl-N, N-dimethyl-N-tempoylammonium bromide spin labels located in different membrane depths. 2-ATDBr was found to be a more potent membrane perturbant than 2-ATDNO both at equal membrane and sample concentrations; this is in compliance with the respective antimicrobial activities of these agents. Using the statistical model of hydrocarbon chains in lipid bilayers, the probabilities of the formation of gauche conformations and the effective energy differences between the trans and gauche conformations were calculated from m-DSA order parameters for two different bilayer regions. Based on these parameters, a molecular model of the location of surfactant molecules in bilayer has been formulated. It has been suggested that at low concentrations the surfactant molecules are located in structural defects between lipid clusters in the bilayer. After filling up these defects, the surfactant molecules penetrate into the clusters between lipid molecules, expand the bilayer laterally and increase the amount of gauche conformations in the hydrocarbon chains in the hydrophobic core of the bilayer. PMID- 2550315 TI - Is Ca2+ antagonists binding protein from cytosolic fraction the precursor of alpha 1-subunit of Ca2+ channel? AB - The binding of Ca2+ antagonists to soluble proteins obtained by ammonium sulphate precipitation from cytosol fraction of rabbit skeletal muscles was studied. The KD values for 3H D-888 and 3H PN 200-110 binding to soluble proteins were 21.3 +/ 3.1 nmol.l-1 and 28.8 +/- 8.9 nmol.l-1 respectively. Photoaffinity labelling of the soluble proteins with the arylazide 1,4-dihydropyridine probe 3H azidopine resulted in labelling of the 85-95 K protein band as determined by SDS polyacrylamide gel electrophoresis. Partial purification of prelabelled soluble sample by gel filtration on Sephadex G-150 gave a more precise molecular weight of 90 +/- 2.5K. Polyclonal antibodies prepared against Ca2+ channel complex from rabbit muscle T-tubules inhibited the 3H PN 200-110 binding. Our results suggest that the soluble protein with Mr = 90K +/- 2.5K may be a precursor of the large subunit of the membrane bound L-type Ca2+ channel in rabbit skeletal muscle. PMID- 2550316 TI - Transcription initiation and a RNA polymerase binding site upstream of the purE gene of the archaebacterium Methanobacterium thermoautotrophicum strain delta H. AB - DNA-dependent RNA-polymerase (RNAP) purified from the thermophilic archaebacterium Methanobacterium thermoautotrophicum strain delta H has been shown to bind specifically to DNA in the intergenic region upstream of the purE gene cloned from this species. The RNAP binding site has been limited to a 41 bp region of DNA which contains the Box A archaebacterial promoter sequence, 5'ATTAAATA. Transcription of the purE gene appears to initiate in vivo at three locations 20-22 bp downstream of the Box A sequence and 27-29 bp upstream of the ATG translation initiation codon of the purE gene. PMID- 2550317 TI - Transformation and expression of a cloned delta-endotoxin gene in Bacillus thuringiensis. AB - A shuttle vector containing the replication region of a resident plasmid of B. thuringiensis, was used to determine the conditions allowing efficient transformation of B. thuringiensis by electroporation. Using this plasmid a delta endotoxin gene was cloned and expressed both in Escherichia coli and B. thuringiensis. It was shown that this gene was poorly expressed in the wild type situation whereas after cloning in acrystalliferous strains of B. thuringiensis large amounts of crystal protein were obtained. PMID- 2550318 TI - A simplified method for the characterization of nuclear polyhedrosis virus genomes. AB - A simplified restriction endonuclease analysis procedure is described which allows the characterization of baculovirus DNA obtained directly from a single larvae without purification of virus. This rapid method was used to demonstrate the genomic stability of nuclear polyhedrosis viruses (NPVs) from Agrotis segetum, Euxoa messoria and Mamestra brassicae after several passages in Euxoa scandens. PMID- 2550320 TI - An adenovirus early region 4 gene product is required for induction of the infection-specific form of cellular E2F activity. AB - E2F is a cellular, sequence-specific DNA-binding factor that binds to pairs of sites that occur upstream of the E1A and E2 early mRNA cap sites. During adenovirus infection, there is induction of a form of E2F that binds cooperatively to the pair of sites in the E2 control region. Production of the infection-specific E2F activity is dependent on early region 4 (E4), as extracts of cells infected with a mutant that lacks E4 did not contain this activity. Instead, two new forms of E2F were seen with the E4 mutant. Infection with mutant viruses unable to make E1A gene products produced the wild-type infection specific E2F activity after a delay. Mutations in the E1B-55 kD-, E1B-21 kD-, E2 72 kD-, and E3-coding regions had no effect on production of infection-specific E2F. Analysis of cell lines confirmed the results obtained with mutant viruses. Cells that expressed E1A but not E4 genes (e.g., 293 cells) did not contain infection-specific E2F. Cell lines that expressed the E4 gene contained the activity. These observations demonstrate that E4 participates in the infection induced change in E2F-binding activity. The data are consistent with E1A playing an indirect role in the process by mediating the efficient expression of E4 gene products which, in turn, induce the alteration in E2F activity. PMID- 2550319 TI - A cellular protein that binds to the 5'-noncoding region of poliovirus RNA: implications for internal translation initiation. AB - Initiation of translation on poliovirus mRNA occurs by internal binding of ribosomes to a region within the 5'-noncoding portion of the mRNA. The mechanistic details and trans-acting factors involved in this event are not understood fully. We used a mobility-shift electrophoresis assay to identify a specific RNA-protein complex, which can form between an RNA fragment that contains nucleotides 559-624 of the poliovirus 5' UTR (untranslated region) and a component or components of a HeLa cell extract. Complex formation was reduced greatly in a reticulocyte lysate or a wheat-germ extract. A 52-kD polypeptide (p52) has been identified as part of the protein-RNA complex by use of an UV cross-linking assay. This polypeptide apparently is not a known translation initiation or elongation factor. The possible involvement of p52 in translation initiation of poliovirus protein synthesis is discussed. PMID- 2550321 TI - Perturbed development of the mouse lens by polyomavirus large T antigen does not lead to tumor formation. AB - To study how the oncogenic process may involve effects on differentiation, we overexpressed an immortalizing oncogene in a developing tissue in transgenic mice. By use of a gene fusion of the alpha A-crystallin promoter to the viral immortalizing oncogene, polyoma large T antigen (PyLT), we created transgenic mice that express PyLT specifically in ocular lens. Expression of large T antigen during embryonic development led to a perturbation in lens development, specifically, an interference with the normal program of fiber cell differentiation. This resulted in microphthalmia, which persisted throughout the life of the animal. Histological analysis revealed impairment of cell elongation, denucleation, and mitotic senescence in both primary and secondary fiber cell differentiation. Strikingly, there was no evidence for hyperplasia or for tumor development in vivo, unlike the consequences of many immortalizing oncogenes on tissues in other transgenic mice. In vitro, however, the developmentally perturbed cells derived from the transgenic lens showed high proliferative capacity. Our results suggest that a primary effect of aberrant expression of an immortalizing gene is an interference with normal tissue development; however, this interference may not necessarily induce proliferation or lead to tumor formation. PMID- 2550322 TI - Isolation and characterization of a gene specifically expressed in different metastatic cells and whose deduced gene product has a high degree of homology to a Ca2+-binding protein family. AB - The gene mts1, which is expressed specifically in metastatic cells, was isolated by molecular cloning coupled with differential DNA reassociation. Transcription of mts1 was found not only in tumor cells, but also in normal cells; homologous RNA was detected only in spleen, thymus, bone marrow, and blood lymphocytes. DNA sequencing of mts1 revealed an open reading frame containing information for a peptide of 101 amino acids, and the amino acid sequence suggested that the mts1 protein was identical to the previously isolated Ca2+-binding mouse protein (Jackson-Grusby et al. 1987; Goto et al. 1988). Thus, the mts1 protein is a member of the calcium-modulated protein family, and our data indicate that mts1 is involved in regulating the metastatic behavior of tumor cells. PMID- 2550323 TI - The yeast transcription activator PRTF, a homolog of the mammalian serum response factor, is encoded by the MCM1 gene. AB - Two proteins, alpha 1 and pheromone/receptor transcription factor (PRTF), bind cooperatively to the upstream activation sequences (UAS) of yeast alpha-specific genes and thereby activate their transcription. In these protein-DNA complexes, the PRTF moiety interacts with a degenerate dyad symmetric sequence, the P box. PRTF contributes also to the regulation of a second set of cell-type-specific genes, the a-specific genes. We used two in vitro assays to show that PRTF is encoded, at least in part, by the MCM1 gene. In one assay, truncated MCM1 proteins encoded by deletion derivatives of the MCM1 gene formed protein-DNA complexes of novel mobility, demonstrated that MCM1 can bind to the P-box containing DNA. Second, antibodies raised to a synthetic MCM1 polypeptide retard the migration of PRTF-DNA complexes in gel mobility shift assays. This result indicates that PRTF, defined as an activity that binds cooperatively with alpha 1 to alpha-specific UAS elements, shares an epitope with MCM1. In addition, we show that MCM1 deletions that remove the carboxy-terminal 129 codons of 286 total codons encode truncated MCM1 molecules that are competent to activate transcription in vivo, indicating that the carboxy-terminal residues are not required for this process. PMID- 2550324 TI - A pituitary POU domain protein, Pit-1, activates both growth hormone and prolactin promoters transcriptionally. AB - The anterior pituitary gland provides a model for investigating the molecular basis for the appearance of phenotypically distinct cell types within an organ, a central question in development. The rat prolactin and growth hormone genes are expressed selectively in distinct cell types (lactotrophs and somatotrophs, respectively) of the anterior pituitary gland, reflecting differential mechanisms of gene activation or restriction, as a result of the interactions of multiple factors binding to these genes. We find that when the pituitary-specific 33-kD transcription factor Pit-1, expressed normally in both lactotrophs and somatotrophs, is expressed in either the heterologous HeLa cell line or in bacteria, it binds to and activates transcription from both growth hormone and prolactin promoters in vitro at levels even 10-fold lower than those normally present in pituitary cells. This suggests that a single factor, Pit-1, may be capable of activating the expression of two genes that define different anterior pituitary cell phenotypes. Because a putative lactotroph cell line (235-1) that does not express the growth hormone gene, but only the prolactin gene, appears to contain high levels of functional Pit-1, a mechanism selectively preventing growth hormone gene expression may, in part, account for the lactotroph phenotype. PMID- 2550325 TI - Efficient synthesis of influenza virus hemagglutinin in mammalian cells with an extrachromosomal Epstein-Barr virus vector. AB - The capability of an Epstein-Barr virus hybrid vector (EBV-CMV), containing the cytomegalovirus (CMV) immediate early enhancer and simian virus 40 promoter, to produce large amounts of authentic mammalian proteins was studied. The cDNA of influenza virus hemagglutinin (HA), a cell surface glycoprotein, was inserted into this vector and the EBV-CMV-HA plasmid was transfected into two human and two monkey cell lines. Southern-blot analysis revealed that the EBV-CMV-HA plasmid was maintained in extrachromosomal state and the recombinant cell clones contained on the average three copies (range 1-24) of the transfected DNA. The recombinant HA polypeptides from different cell clones, selected either randomly or by fluorescence-activated cell sorter, were analysed using immunological techniques. Three of the four cell lines expressed recombinant HA on the cell surface in glycosylated form. The highest production levels, 11.5 micrograms/10(6) cells, were obtained in HeLa cells containing only two copies of EBV-CMV-HA DNA per cell. The protein levels correlated with the mRNA levels in Northern-blot analysis. A corresponding vector, containing the same regulatory signals for HA expression, but lacking the EBV sequences, yielded clones with significantly lower expression levels. The results confirm that the extrachromosomal EBV-CMV vector is very useful in the production of apparently authentic mammalian glycoproteins. PMID- 2550326 TI - Characterization of the plasmid genes blaT-4 and blaT-5 which encode the broad spectrum beta-lactamases TEM-4 and TEM-5 in enterobacteriaceae. AB - We have determined the nucleotide sequence of the plasmid genes blaT-4 and blaT-5 which encode the broad-substrate-range beta-lactamases TEM-4 and TEM-5, respectively. The TEM-4 enzyme, which confers high-level resistance to cefotaxime (Ctx) and ceftazidime (Caz), differed from the TEM-1 penicillinase by four amino acid substitutions. Two of the mutations are identical to those responsible for the wide substrate range of the TEM-3 beta-lactamase which hydrolyses Ctx and Caz. The amino acid sequence of TEM-5, which confers higher levels of resistance to Caz than to other recently developed cephalosporins, differed from that of TEM 1 by three mutations distinct from those of TEM-4. Analysis of the location of the mutations in the primary and tertiary structures of class A beta-lactamases suggests that interactions between the substituted residues and beta-lactam antibiotics non-hydrolysable by TEM-1 and TEM-2 allow TEM-4 and TEM-5 to hydrolyse efficiently novel broad-spectrum cephalosporins such as Ctx and Caz. PMID- 2550327 TI - Nucleotide sequence of the chromosomal gene coding for the aminoglycoside 6 adenylyltransferase from Bacillus subtilis Marburg 168. AB - Gene aadK of Bacillus subtilis is 855 bp long and codes for aminoglycoside 6 adenylyltransferase. PMID- 2550328 TI - Gene dosage effect on the expression of the delta-endotoxin genes of Bacillus thuringiensis subsp. kurstaki in Bacillus subtilis and Bacillus megaterium. AB - Significant differences in expression of the delta-endotoxin genes cryA1 and cryA2 of Bacillus thuringiensis subsp. kurstaki were observed in B. subtilis and B. megaterium. The cryA1 gene was expressed when present on a high-copy-number (hcn) vector in B. megaterium but not in B. subtilis. The cryA2 gene was expressed in both hosts, but at a higher level in B. megaterium. Expression of the cryA2 gene in B. megaterium was better from a hcn vector than from a low copy number vector. Inhibition of sporulation was observed when the toxin genes were present on hcn plasmids in B. subtilis while no such effect was evident in B. megaterium. In addition, there was a significant reduction in copy numbers in both B. subtilis and B. megaterium when delta-endotoxin genes or a spoVG promoter containing fragment of DNA were cloned into hcn plasmids. PMID- 2550329 TI - Cloning and characterization of a phosphate-regulated promoter involved in phosphate control of candicidin biosynthesis. AB - Phosphate strongly repressed the formation of p-aminobenzoic acid (PABA) synthase, an enzyme involved in candicidin biosynthesis. Expression in Streptomyces lividans of the pabS gene (encoding PABA synthase) of Streptomyces griseus is repressed by phosphate at concentrations above 0.1 mM. However, expression of the pabS gene in Escherichia coli is not regulated by phosphate. Phosphate control of the expression of the pabS gene was observed in all plasmids containing the original 4.5-kb BamHI fragment, whereas no phosphate regulation was found when an upstream 1-kb fragment that carries the pabS promoter was deleted. Using the promoter-probe plasmid pIJ424, a '114-bp' promoter was cloned. Expression of the promoterless kanamycin phosphotransferase gene when fused to the '114-bp' promoter was strongly reduced by phosphate (90% at 5 mM concentration). The '114-bp' promoter has been sequenced and the first transcribed nucleotide identified by S1 mapping. The '114-bp' fragment is A + T rich (54%), as compared to the Streptomyces genome (70-73% GC). The presence of a phosphate control sequence (pcs) in the upstream region of the pabS gene is proposed. PMID- 2550330 TI - Radicals involved in photoallergen/protein interactions. AB - Aqueous solutions (pH = 8) of both 3,3'-dimethyl and 4,4'-dimethyl substituted analogues of the photoallergen fentichlor (bis(2-hydroxy-5-chlorophenyl)sulphide) produced stable semiquinone radicals when irradiated with u.v. light (greater than 310 nm). These radicals have been characterised using electron spin resonance techniques: the results confirm the assignment of hyperfine coupling constants for the parent fentichlor radical. The binding of fentichlor to HSA was found to be partly oxygen dependent demonstrating a role for semiquinone type radicals in the binding mechanism. The stoichiometry and specificity of the binding of the dimethyl analogues to soluble proteins were found to be similar to that of fentichlor itself. PMID- 2550331 TI - Free radical lipid oxidation affects cholesterol transfer between lipoproteins and erythrocytes. AB - Human erythrocytes were incubated for 5 h at 37 degrees C with lipoproteins (LP), preliminary oxidized to different extent, as assessed by thiobarbituric acid (TBA) test. Cholesterol content in the cells was increased by 12-14% after incubation with low-density lipoproteins (LDL) along with augmentation of order parameter and rotational correlation time of spin-labeled stearic acids incorporated into membranes. If erythrocytes were incubated with oxidized LDL, containing 2.5-4 times more TBA-reactive material than native ones, cellular content of cholesterol was increased by 24-28%. In contrast, high-density lipoproteins (HDL2 and HDL3) removed cholesterol from cell membranes, when incubated with erythrocytes. This was followed by increased fluidity of membrane lipid phase as detected by the spin probe method. Oxidation of HDL2 and HDL3 decreased their ability to accept cholesterol from cell membranes. No detectable accumulation of TBA-reactive material was observed in the samples during the incubation. The antioxidant, butylated hydroxytoluene (BHT), in the concentration of 10(-5) M did not influence the cholesterol transfer between LP and erythrocytes. Hence, the effects of lipid peroxidation (LPO) on the cholesterol transfer seem to result from LP alterations by oxidation rather than from free radical reactions occurring during the incubation. By increasing cholesterol donating ability of LDL and inhibition of cholesterol-accepting capacity of HDL lipid peroxidation in LP may activate cholesterol accumulation in blood vessel cells and thus contribute to atherosclerosis. PMID- 2550332 TI - Sonochemistry of nitrone spin traps in aqueous solutions. Evidence for pyrolysis radicals from spin traps. AB - When argon-saturated aqueous solutions of alpha-phenyl-N-tert-butylnitrone (PBN) were sonicated, the spin adducts PBN-Phenyl (Ph), PBN-X, and PBN-H were observed. It can be inferred that PBN-Ph and -X arise from spin adducts of thermal decomposition products of PBN induced by the high temperature due to ultrasonic cavitation. The ESR signal of PBN-H was observed at a lower PBN concentration than those of PBN-Ph and PBN-X. The ratios of ESR intensity of PBN-H to those of PBN-Ph and PBN-X increased with the final temperatures of the cavitation bubbles created by different rare gases. The spin adducts of methyl and tert-butyl radicals from the pyrolysis of PBN, induced by the high temperatures due to cavitation, were found from spin trapping experiments in which 3,5-dibromo-2,6 dideuterio-4-nitrosobenzene sulfonate was used as a spin trap. Similar spin adducts induced by pyrolysis were also observed in sonicated aqueous solutions of other nitrone spin traps, such as alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone, and alpha-(4-nitrophenol) N-tert-butylnitrone. The greater the hydrophobicity of the spin traps, as measured by the 2-octanol/water partition coefficients, the lower the concentration of spin trap at which methyl radicals generated by thermal decomposition of the spin trap can be observed. The present results indicate that the nonvolatile, highly hydrophobic spin traps accumulate preferentially in the interfacial region of cavitation bubbles where they undergo thermal decomposition during cavitation to produce the radicals. PMID- 2550333 TI - Oxygen-concentration dependence of microsomal chemiluminescence. AB - The effect of varying concentrations of oxygen on NADPH-dependent microsomal chemiluminescence was determined. Light emission increased as the concentration of oxygen was elevated from 0 to 10 to 20%, and then began to decrease upon further increases in oxygen concentration to 50 and 100%. This biphasic response of chemiluminescence is similar to that previously observed for microsomal generation of hydroxyl radical, however, the light emission was not sensitive to superoxide dismutase, catalase or benzoate confirming the lack of a role for .OH in the light emission. The biphasic nature of the response of chemiluminescence is similar to that reported for exhalation of ethane and pentane but not that of malondialdehyde as a measure of lipid peroxidation, although the concentrations of O2 to reach the maximum effect differ. Activity of NADPH-cytochrome P450 reductase was decreased at the elevated concentrations of O2. The biphasic response of chemiluminescence to O2 appears to reflect the need for a critical amount of O2 to generate the initiating oxidizing species, and the effect of O2 on the appropriate redox state of the iron catalyst. PMID- 2550334 TI - Properties of enzymes in hepatocytes that convert 5-HPETE or LTA4 into LTB4. AB - Rat hepatocyte homogenates convert 5-hydroperoxyeicosatetraenoic acid (5-HPETE) into biologically active leukotriene B4 (LTB4) as well as less active all-trans LTB4 (i.e., 6-trans-LTB4 and 6-trans-12-epi-LTB4). Here, we present a hypothesis of the reaction mechanism and the minimal structural requirements of the active enzyme based on the following experimental evidence: The ED50 of the inhibitors 5,8,11,14-eicosatetraynoic acid (ETYA) and 5,6-dehydro-eicosatetraenoic acid was approximately 100-fold higher than for 5-lipoxygenase. Propanethiol and O2 were strong inhibitors of LTB4 formation, whereas butylated hydroxytoluene, nordihydroguaiaretic acid, metyrapone, Desferal and CO had no effect. Cytochrome c, catalase, hematin, and a Fe3+/Fe2+ couple, but not iron-free protoporphyrin IX, catalyzed the formation of only all-trans-LTB4. LTB4 formation in hepatocyte homogenates was heat- and trypsin-sensitive whereas all-trans-LTB4 formation was not. We propose that a ferric heme iron forms a ferryl-hydroxo complex upon homolytic scission of the oxygen-oxygen bond in 5-HPETE and the resulting 5,6 trans-epoxide radical is oxidized by the ferryl-hydroxo complex to yield LTA4. A mechanism for hydrolysis of LTA4 is described that results in formation of LTB4 (less than 1% yield) rather than all-trans-LTB4. PMID- 2550335 TI - Prophylaxis of deep venous thrombosis with a low-molecular-weight heparin (Kabi 2165/Fragmin) in stroke patients. AB - In a group of 60 patients in the acute phase of an ischemic stroke the assumption that low-molecular-weight heparin Kabi 2165 in the dose of 2 X 2,500 anti-Xa units s.c. could prevent thromboembolic complications was tested using a double blind, placebo-controlled, randomized trial design. Thirty patients were allocated to each group. Both treatment groups were comparable with regard to neurological status and general condition. In the Kabi 2165 group there were 6 cases of deep venous thrombosis (DVT) compared to 15 in the placebo group (p = 0.05). In the placebo group there were 4 deaths during the trial versus 9 in the Kabi-2165-treated group (NS). Cerebral bleeding complicated 2 cases in the placebo group versus 4 in the Kabi 2165 group (NS). These results indicate that in ischemic stroke patients Kabi 2165 2 X 2,500 anti-Xa units s.c./24 h reduces the frequency of DVT. Because of the small number of patients it is impossible to evaluate the safety. PMID- 2550336 TI - Metabolism of linamarin in rats. AB - The metabolism of linamarin [2(beta-D-glucopyranosyloxy)isobutyronitrile] was investigated in male albino Wistar rats and using rat liver microsomal preparations. In the in vitro experiments incubations of varying concentrations of linamarin at pH 6.0-6.5 with liver microsomal preparations resulted in rapid degradation of the substrate without concomitant production of any detectable amount of hydrogen cyanide (HCN) or of thiocyanate, its detoxication derivative. Boiled incubation medium did not degrade linamarin. Mathematical treatment of the degradation data generated theoretical HCN values that were used to construct a Lineweaver-Burke plot, which gave apparent Km and Vmax values of 3.3 mM-linamarin and 0.017 mg HCN/min/mg protein, respectively. In the in vivo experiments excretion of glucosidic cyanide (linamarin) in rat urine was found, within the range of applied oral doses 10-350 mg/kg body weight, to be dose dependent. Urinary excretion of HCN and thiocyanate did not show this correlation. Following administration (iv) of 10, 50 or 100 mg linamarin, elimination of the test substance from rat blood was observed to occur exponentially, and the half-life was estimated at about 90 min for all three dose levels. PMID- 2550338 TI - Modulation of phagocytic activity in cultured Sertoli cells. AB - Phagocytic activity of rat Sertoli cells that were cultured in vitro has been evaluated as the ability to internalize polystyrene beads. Our data demonstrate that these cells are active phagocytes and that such phagocytic activity is under negative control by follicle-stimulating hormone (FSH). The hormonal control is mediated by increased intracellular levels of cAMP. Moreover Sertoli cell responds to tuftsin, an oligopeptide known to act only on macrophages and granulocytes, by increasing up to five times its phagocytic activity. Phagocytic uptake of polystyrene beads is associated with dramatic changes of the cellular shape. Such morphological modifications are significantly reduced under FSH stimulation. The physiological implications of the data are discussed. PMID- 2550337 TI - Activation of Ca2+ channels during the acrosome reaction of sea urchin sperm is inhibited by inhibitors of chymotrypsin-like proteases. AB - Probable participation of sperm protease in the acrosome reaction was investigated using several inhibitors and substrates. Among those examined, L-1 tosylamide-2-phenylethyl chloromethyl ketone (TPCK) and chymostatin, chymotrypsin inhibitors, p-nitrophenyl-p'-guanidinobenzoate (NPGB), a serine protease inhibitor, and N-benzoyl-L-tyrosine ethyl ester (BTEE), a chymotrypsin substrate, inhibited the egg jelly-induced acrosome reaction of Strongylocentrotus intermedius. TPCK and BTEE, however, did not inhibit the reaction caused by ionophores, A23187, or nigericin. To know the mechanism of inhibition by chymotrypsin inhibitors and substrates of the egg jelly-induced acrosome reaction, intracellular Ca2+ concentration [( Ca2+]i) and pH (pHi) were measured with fura-2 and 2',7'-bis (carboxy-ethyl)carboxyfluorescein (BCECF), respectively. Egg jelly caused increase of [Ca2+]i, which was depressed by BTEE. Egg jelly also caused a transient rise of pHi, which was not depressed by BTEE. In the presence of verapamil, the acrosome reaction by egg jelly was significantly inhibited concomitant with depressed increase of [Ca2+]i. The rise of pHi was not depressed by verapamil. Thus, modes of action of BTEE and of verapamil are similar to each other. Bringing these findings together, the authors present a view that a chymotrypsin-like protease of sea urchin sperm activates verapamil-sensitive Ca2+ channels, which take part in the acrosome reaction. PMID- 2550340 TI - Glycerylphosphorylcholine diesterase activity of uterine fluid in conditions inducing secondary sex ratio change in the rat. AB - Food restriction or decreasing the ratio of [Na+, K+] to [Ca++, Mg++] in the diet of female rats before conception favoured the production of female offspring. Seven days of food deprivation decreased uterine fluid GPC diesterase activity in female rats, whereas long-term food restriction (21 days), rather than decreasing the enzyme activity, apparently stimulated it. Dietary Ca and Mg supplementation, likewise, produced significant decrease in GPC diesterase activity. A significantly positive correlation was observed between the levels of uterine GPC diesterase and secondary sex ratio change, which indicates that these dietary techniques of sex-ratio manipulation may modulate the uterine fluid GPC diesterase activity and influence sex determination in utero. PMID- 2550339 TI - Inhibition of the human sperm acrosome reaction by proteinase inhibitors. AB - The analogue of the second messenger cAMP, dibutyryl cAMP (dbcAMP), was shown to induce the human sperm acrosome reaction to the same extent as calcium ionophore A23187, providing preliminary evidence for the involvement of the adenylate cyclase system in the acrosome reaction (AR) of human spermatozoa. Using the human synchronous acrosome reaction system, proteinase inhibitors were tested for their effect on the dbcAMP-induced human sperm acrosome reaction. The proteinase inhibitor 4'-acetamidophenyl 4-guanidinobenzoate (AGB), an inhibitor of proacrosin activation and of acrosin, when added at either the onset of incubation or to capacitated spermatozoa, 5 min prior to stimulation by dbcAMP, significantly (P less than 0.01) inhibited the acrosome reaction at final concentrations of 1 x 10(-4) M to 1 x 10(-6) M in comparison to dbcAMP treatment alone. At concentrations less than 1 x 10(-6) M, no significant inhibitory effect was seen. Similarly, para-aminobenzamidine (pAB), also an inhibitor of proacrosin activation and of acrosin, significantly (P less than 0.01) inhibited the dbcAMP induced acrosome reaction at final concentrations of 1 x 10(-4) M to 1 x 10(-6) M when added at either the onset of incubation or to capacitated spermatozoa, 5 min prior to stimulation by dbcAMP, in comparison to stimulation by dbcAMP alone. However, at concentrations less than 1 x 10(-6) M, no significant (P greater than 0.05) inhibitory effect was seen. These results indicate that a serine proteinase, most likely acrosin, has a role in the human sperm acrosome reaction and suggest that the enzyme functions after the involvement of the adenylate cyclase system. PMID- 2550341 TI - Lack of effect of insulin or insulin-like growth factor I on the steroid sulfatase activity of human placental cytotrophoblasts. AB - Hyperinsulinemia is known to reduce serum dehydroepiandrosterone sulfate (DHEA-S) levels in normal females. A possible mechanism for this phenomenon would be an insulin-mediated increase in steroid sulfatase activity, with insulin acting either via activation of the insulin receptor or via cross-reaction with the insulin-like growth factor I (IGF-I) receptor. Using a well characterized human cytotrophoblast system, the presence of steroid sulfatase activity in isolated cytotrophoblasts was documented. Half maximal cellular hydrolysis of DHEA-S was observed at a substrate concentration of 9.6-14.5 microM, and maximal hydrolysis at a concentration of 75-100 microM. The hypothesis that insulin increases steroid sulfatase activity was examined by exposing cytotrophoblasts to supraphysiological concentrations of either insulin (2 micrograms/ml) or IGF-I (20 ng/ml) for 24 h and then measuring the rate of DHEA-S hydrolysis. Insulin failed to affect cytotrophoblastic steroid sulfatase activity, irrespective of whether the substrate concentration was 20 microM or 100 microM. IGF-I also exerted no effect on steroid sulfatase activity. These data indicate that neither insulin nor IGF-I affect the steroid sulfatase activity of human cytotrophoblasts. An effect of insulin or IGF-I on the steroid-sulfatase activity of other tissues has not been excluded. These observations suggest that the decline in serum DHEA-S levels during hyperinsulinemia is not mediated via an insulin-induced increase in steroid sulfatase activity. PMID- 2550342 TI - Hormonal regulation of de novo purine biosynthesis rate. A biomathematical approach. Effect of glucocorticoid hormones on the inosinic branch point in vivo. PMID- 2550343 TI - Insulin action is mimicked by polyclonal antireceptor antibodies that activate the insulin receptor tyrosine kinase. AB - The insulin-like properties of anti-insulin receptor antibodies (P95 Ab) that have been characterized as being directed against the receptor beta-subunit, were studied as probes to assess the interrelationship between insulin action and receptor phosphorylation. When tested on intact cells, P95 Ab mimicked insulin effects. On isolated fat cells, they stimulated 2-deoxyglucose (2-DG) transport and lipogenesis and the P95 antibody maximal effects (173 and 232% of the control values, respectively) represented about 50% of the maximal effects elicited by insulin (317 and 475% of the control values). On cultured Zajdela hepatoma cells (ZHC cells), P95 Ab also mimicked insulin action on the incorporation of [U 14C]glucose into glycogen (158 and 207% of the control value for antibody- and insulin-treated cells, respectively). In all cases the antibody effects were dose dependent, specific and, when maximal, were not additive with those elicited by insulin. When tested in a cell-free system, P95 Ab faithfully reproduced insulin action on the phosphorylation of the receptor beta-subunit. The maximal antibody and insulin effects (317 and 328% of the control value, respectively) were not additive. P95 Ab were also equally potent as insulin to stimulate the receptor mediated phosphorylation of an exogenous substrate (365 and 379% of the control value in P95 antibody- and insulin-treated receptors, respectively). As well, P95 Ab proved as able as insulin in stimulating the tyrosine kinase activity of the receptor (89% of the hormone effect) when the activation was carried out in vivo. Taken together, these results are consistent with a role for the kinase activity of the insulin receptor in mediating the action of insulin. PMID- 2550344 TI - Unlike heparin, low-molecular weight heparin does not suppress aldosterone production. PMID- 2550345 TI - Successful endoscopic obturation of gastric varices with butyl cyanoacrylate. AB - In 27 patients who had bled from esophagogastric varices, large-sized and/or actively bleeding gastric varices were endoscopically obturated with the tissue adhesive butyl cyanoacrylate. Active bleeding was stopped in six patients. Rebleeding occurred in 10 patients; in four patients, rebleeding was due to ruptured gastric varices, occurred early and was successfully treated by reinjection of gastric varices; in one patient, rebleeding was attributed to ulceration on an injected gastric varix. Eight patients died: two of rebleeding (from esophageal varices or undetermined source), four of sepsis and/or liver failure and two at home of undetermined cause. No specific complication due to injection of gastric varices was observed. The results obtained in this series of patients with gastric varices obturated by injection of butyl cyanoacrylate are much more satisfactory than those obtained in previously published series of patients with gastric varices treated by injection of sclerosants. PMID- 2550346 TI - Ubiquitin--a common denominator in intermediate filament pathology of brain and liver? PMID- 2550347 TI - Reovirus 3 and neonatal biliary disease: discussion of divergent results. PMID- 2550348 TI - Adenoid cystic carcinoma of the endometrium. AB - An adenoid cystic carcinoma of the endometrium co-existing with an endometrioid adenocarcinoma is presented. Immunohistochemical staining for actin and keratin suggests a myoepithelial differentiation of tumour cells in adenoid cystic carcinoma. PMID- 2550349 TI - Expression of intermediate filaments in malignant fibrous histiocytomas. AB - The expression of intermediate filaments (IFs) in 34 malignant fibrous histiocytomas (MFHs) was studied immunohistochemically and ultrastructurally. Using the avidin-biotin-peroxidase method, positive reactions were detected as follows: for desmin in 12 tumors, for neurofilament in two tumors, for cytokeratin in one tumor, and for vimentin in 30 tumors. Desmin immunoreactivity was found in tumors of all four histologic subtypes and cytokeratin immunoreactivity was found in one tumor of the myxoid type. Because of the cross reactivity of anti-neurofilament antibody with reactive histiocytes, the immunoreactivity for neurofilament seemed to be non-specific. Ultrastructurally, five of 13 tumors studied contained some tumor cells showing myofibroblastic or smooth muscle cell differentiation. A few tumor cells in one cytokeratin-positive tumor had tonofilaments in their cytoplasm. Desmin expression in some MFHs seemed to be due to myofibroblastic or smooth muscle cell differentiation of some tumor cells. Cytokeratin expression seemed to indicate epithelial differentiation in some MFHs. This varied expression of IFs in MFHs may reflect the heterogeneous nature of MFHs, and suggests that MFHs represent the final stages of dedifferentiation of several different types of sarcomas or, alternatively, represent forms of poorly differentiated sarcoma with the potential of developing into more differentiated sarcomas of heterogeneous origin. PMID- 2550351 TI - Ductal adenoma of the breast--a review of fifteen cases. AB - The term ductal adenoma has been recently introduced to describe a solid benign lesion of breast ducts. This study describes the clinical, morphologic, and immunohistochemical features of 15 cases of ductal adenoma. Ductal adenomas are usually single, occasionally multiple, lesions occupying medium- and large-sized breast ducts. They may occur in women of all ages, although the majority of patients are 60 years of age or greater. Ductal adenomas usually present clinically as breast lumps which may mimic carcinoma; less commonly, they are associated with nipple discharge. Patients in this series showed no family or previous history of breast disease and had uneventful follow-up after local excision. Despite often showing worrying pseudoinfiltration and cytologic atypia, the immunohistochemical demonstration of a myoepithelial layer and intact basement membrane around the tubules was clear evidence of the benign nature of the lesions. We conclude that most ductal adenomas evolve by sclerosis of benign intraduct papillary lesions, although processes similar to sclerosing adenosis and, possibly, duct ectasia may contribute to the pathogenesis of a proportion of cases. It is hoped that a wider appreciation of the entity of ductal adenoma will reduce the diagnostic uncertainty that continues to surround these and related lesions. PMID- 2550350 TI - Immunohistochemical analysis of calcitonin and calcitonin gene-related peptide in human lung. AB - Calcitonin and calcitonin gene-related peptide (CGRP) have been localized immunohistochemically in neuroendocrine cells of normal and diseased human lungs. Cells immunoreactive for calcitonin and CGRP first appeared in immature bronchi in the 27th gestational week. Thereafter, both peptides were found in the same bronchial neuroendocrine cells throughout fetal and neonatal life. In adult lungs with or without neuroendocrine cell hyperplasia, only calcitonin was present. In 17 of 18 (94%) pulmonary tumorlets, variable numbers of calcitonin-positive cells were identified. A few CGRP immunoreactive cells, as a subset of calcitonin containing cells, were found in only three (17%) lesions. Of 37 bronchial carcinoids, calcitonin was detected in 14 and CGRP was detected in 16 (38% and 43%, respectively), and both peptides were predominantly localized in the same cells. Ten of 45 (22%) small cell lung carcinomas were calcitonin-immunoreactive. CGRP was noted in only one (2%) of these tumors, and both peptides coexisted in single cells. These findings indicate that the patterns of calcitonin/CGRP expression in hyperplastic bronchial neuroendocrine cells, pulmonary tumorlets, and, to some extent, small cell lung carcinomas are similar to those of normal adult lungs. On the other hand, calcitonin/CGRP expression in bronchial carcinoids is similar to that of late fetal and neonatal lungs. PMID- 2550353 TI - Localization of polymorphic DNA probes frequently deleted in lung carcinoma. AB - Five polymorphic DNA segments from human chromosome 3, that are frequently deleted in lung carcinoma were mapped by non-isotopic in situ hybridization to metaphase chromosomes. The DNA segment D3S3 mapped to 3p13-p14.2, D3S6 to 3p14.3 p14.5, D3S48 to distal 3p21-p22, ERBA beta to 3p24.3 and ERBA2 to 3p24.3. The map location of ERBA beta and ERBA2 was confirmed by re-mapping each probe in combination with D3S6 as a marker for 3p14. PMID- 2550352 TI - Characterization of patients with glycerol kinase deficiency utilizing cDNA probes for the Duchenne muscular dystrophy locus. AB - Genomic DNA from five previously unreported patients with glycerol kinase deficiency (GKD), dystrophic myopathy, and adrenal insufficiency were studied with genomic probes and cDNA probes for the Duchenne muscular dystrophy (DMD) locus. These individuals, together with those reported by ourselves and others, show that patients with a contiguous gene syndrome involving the DMD, GK, and adrenal hypoplasia congenita (AHC) loci have a broader distribution of microdeletion breakpoints than those observed among patients with classical DMD. This study demonstrates the use of the DMD cDNA probes to delineate the centromeric deletion breakpoints for patients with Xp21 microdeletions extending beyond the DMD locus. It also shows the practical diagnostic application of the DMD cDNA probes when the diagnosis of GKD is entertained in a patient with known DMD and only DNA is available for study. PMID- 2550354 TI - Epigenetic changes may contribute to the formation and spontaneous regression of retinoblastoma. AB - Epigenetic models for tumor formation assume that oncogenic transformation results from changes in the activity of otherwise normal genes. Since gene activity can be inhibited by DNA methylation, and inactivation of tumor suppressor genes is a fundamental process in oncogenesis, we investigated the methylation status of the retinoblastoma suppressor gene (RB gene) on chromosome 13, in blood and tumor cells from 21 retinoblastoma patients. Using methylation sensitive restriction enzymes and a cloned DNA probe for the unmethylated CpG island at the 5' end of RB gene, we obtained evidence of hypermethylation of this gene in a sporadic unilateral retinoblastoma tumor. The closely linked esterase D gene and a CpG-rich island on chromosome 15 were not affected. We suggest that changes in the methylation pattern of the RB gene play a role in the development and spontaneous regression of some retinoblastoma tumors. PMID- 2550355 TI - Regional localization of human ecto-5' nucleotidase to chromosome 6q14-q21. AB - Human and mouse hybrids that contain fragments of human chromosome 6 as translocations were analysed for expression of ecto-5' nucleotidase enzymic activity measured by the conversion of AMP to adenosine and for antigenicity recognized by a monoclonal antibody specific for the human isozyme. Both methods allow a regional assignment of ecto-5'nucleotidase to 6q14-q21. PMID- 2550356 TI - The human homolog of the myeloproliferative virus maps to chromosome band 1p34. AB - The human homologue of the recently isolated myeloproliferative leukemia virus, a retrovirus that induces myeloproliferative disorder in mouse, has been mapped in man to chromosome band 1p34 by in situ hybridization. PMID- 2550357 TI - Chronic neurological toxicity associated with exposure to volatile substances. AB - 1. The main neurological disorders associated with chronic VSA are peripheral neuropathy, cerebellar disease, chronic encephalopathy and dementia. Apart from peripheral neuropathy, the clinical features are non-specific, evidence for solvent-related toxicity is in most cases circumstantial and there is no clear dose/response relationship. 2. Peripheral neuropathy is mainly associated with n hexane and methyl n-butyl ketone. 3. Cerebellar disease is usually associated with toluene exposure; in the more severe cases there is often radiological evidence of irreversible cerebellar atrophy. 4. Chronic encephalopathy and dementia are the most serious consequence of solvent exposure, particularly to toluene in abusers and to mixed solvents in industrial workers. Postmortem studies in some abusers have shown generalized axonal degeneration, demyelination and brain atrophy. 5. Further studies on low level solvent exposure are needed as little is known about the neurological consequences of mild VSA, especially as regards individual susceptibility and possible interactions between solvents and other toxins such as ethanol. PMID- 2550358 TI - Stability of specific immunoglobulin secretion by EBV-transformed lymphoblastoid cells and human-murine heterohybridomas. AB - We have examined variables leading to the generation of stable, antigen-specific, human immunoglobulin-secreting cell lines. Peripheral blood B lymphocytes enriched for Thomsen-Friedenreich antigen (T antigen)-specific cells were transformed with Epstein-Barr virus. Lymphoblastoid cells (LC) reactive with T antigen were either expanded without cloning or cloned at limiting dilution and then fused with murine 653 cells. Uncloned LCs from three transformations secreting polyclonal anti-T antibody (7-18 micrograms/ml/10(6) cells/24 hr total immunoglobulin) were subcultured at 100 cells/well, and T antigen-reactive cultures pooled. These cultures quickly lost specific antibody secretion, presumably due to overgrowth by clones of unknown specificity. T antigen-reactive LCs that were cloned three times at limiting dilution secreted 0.2 - 6.1 micrograms/ml/10(6) cells/24 hr but died or stopped secreting specific immunoglobulin after 77 to 155 days in culture. Pooling T antigen-reactive clones after each cloning step did not increase the long term stability compared to unpooled clones (p = 0.2). Fusions between cloned LCs and 653 cells failed to yield viable hybrids in nine of ten attempts with seven different LC lines. In contrast, fusion of uncloned LCs and 653 cells resulted in the generation of viable immunoglobulin-secreting heterohybrids in 22 of 24 fusions. The heterohybridomas produced from fusion of uncloned T antigen-reactive cultures with 653 cells secreted significantly more antibody (frequency of cell lines secreting greater than 2 micrograms/ml/10(6) cells/24 hr, p less than 0.01) and higher titers of antibody (frequency of cell lines secreting greater than four hemagglutination units of T antigen-specific antibody, p less than 0.03) than cloned lymphoblastoid cells. The hybrids maintained specific immunoglobulin secretion for longer in culture than either cloned or uncloned lymphoblastoid cell lines (p less than 0.001). PMID- 2550359 TI - Monoclonal antibodies M340 and UJ181.4 recognize antigens associated with primitive neuroectodermal tumours/tissues. AB - Monoclonal antibodies UJ181.4 and M340 were raised from separate fusions using either human foetal brain or medulloblastoma tissue as immunogens. The antibodies are both of IgG isotype and bind to their target antigens with affinities in excess of 10(-9) M. Both monoclonal antibodies have been shown to bind to primitive neuroectodermal tumours and human foetal brain. Expression of the two antigens is developmentally regulated as no binding is detected on adult brain as determined by a variety of indirect binding assays. The two monoclonal antibodies can be clearly distinguished by their migration in a pH gradient (isoelectric focussing gels). UJ181.4 and M340 clearly recognize two different epitopes, as it is not possible to block the binding of one antibody with an excess of the other. In addition, careful study of the binding profiles of the two reagents suggest that they recognize two distinct antigens, although these have not been biochemically characterized. The reagents have proved particularly useful in both the differential diagnosis of the small round cell tumours of childhood and anaplastic brain tumours. In addition, they have formed part of a panel of reagents used for the immunomagnetic depletion of neuroblasts from bone marrow and have been used for targeting radionuclides to tumour cells in the intrathecal compartment. PMID- 2550360 TI - Effect of dimethyl sulphoxide on mitochondrial biogenesis in regenerating rat liver and Saccharomyces cerevisiae. AB - Effect of dimethyl sulphoxide (DMSO) on mitochondrial biogenesis in regenerating rat liver and cells of Saccharomyces cerevisiae during aerobiosis has been studied by monitoring the cytochrome oxidase activity. A single dose of DMSO (275 mg/100-125 g body wt) to normal rats stimulated cytochrome oxidase activity in liver mitochondria while the same dose to partial hepatectomized rats inhibited the enzyme activity. Administration of low dose of DMSO (92 mg/100-125 g body wt) to partial hepatectomized rats did not alter the enzyme activity. Anaerobic cells of S. cerevisiae on aerobiosis for 2 hr attained cytochrome oxidase activity level on par with aerobic cells. Inclusion of DMSO (275 mg/100 ml) in the growth medium of S. cerevisiae during respiratory adaptation exerted partial inhibitory effect on the formation of cytochrome oxidase at 2 hr period, while the 10-fold concentration inhibited the enzyme formation completely. However, the inhibitory effect of DMSO on enzyme formation was abolished on prolonged growth (18 hr and above), while these doses had no influence on cytochrome oxidase in aerobic cells of S. cerevisiae. The results imply that DMSO may be exerting its effect on the assembly of subunits into active enzyme complex during mitochondrial biogenesis. PMID- 2550361 TI - Management of carcinoma breast with conservative surgery and radiation therapy experience in a developing country. AB - Between 1980 and 1987, 14 patients with breast cancer who refused mastectomy were treated at the V.N. Cancer Centre, Coimbatore, with excision of the lump, followed by radiation therapy and adjuvant chemotherapy/hormonal therapy. No attempt was made to boost the tumour bed with interstitial implants or electron beam for want of these facilities. No patient developed local or loco-regional disease or distant metastases during the period of follow-up which ranged between 10 and 76 months. Ten of the 14 patients have been followed up for more than two years. Most published data on breast-conserving treatment for cancer breast are from the U.S., UK and Europe. There are not many papers published from the developing countries. This paper gives an insight into this subject in the developing countries. PMID- 2550363 TI - Protective immunity induced by porin against Salmonella infection in mice. AB - Porin, a major outer membrane protein was purified from Salmonella typhimurium and its immune potential was studied in mice. Active immunization with porin induced about 45 per cent protection to an intravenous challenge with 10LD50 of S. typhimurium. Further, in porin immunized mice significant level of anti-porin antibodies and DTH reaction were detected. Attempts were also made to improve the immune potential of porin. Freund's complete adjuvant when mixed with immunogenic doses of porin enhanced the anti-porin antibody titre. However, it could not improve the protective ability of porin. On the other hand, porin when injected along with lipopolysaccharide (LPS) induced a higher level (55% survival with 50LD50) of protection than porin or LPS alone. This finding was also substantiated by the significantly reduced in vivo growth of challenge organisms in mice immunized with porin plus LPS. These results indicate that porin is a protective antigen and LPS significantly enhances the protective ability of porin. PMID- 2550362 TI - FN-C1q and C1 INH C1r-C1s complexes as indicators of complement activation in patients with chronic lymphocytic leukaemia. AB - We have previously found low levels of C1 and C4 INH in the sera of chronic lymphocytic leukaemia (CLL) patients. Hypocomplementaemia was supposed to be the consequence of a permanent activation of the classical pathway. We have compared the levels of C1 INH-C1rC1s and C1q-FN complexes in the sera of 95 CLL patients and 100 healthy controls, because these complexes are known to be formed in the early stage of classical pathway activation. A significant increase in the level of both types of complexes was found in sera of CLL patients as compared to the controls. These findings support the assumption that the classical complement pathway is activated in the patients with CLL. PMID- 2550364 TI - Evaluation of a new latex agglutination kit for detection of human rotavirus in faecal specimens. AB - A commercial latex agglutination (LA) test was compared with ELISA and direct electron microscopy (EM) for detection of rotavirus antigen in 93 stool specimens obtained from as many children with acute gastroenteritis. Seventy one specimens (76.3%) were either positive or negative with all the three techniques, while 22 (23.7%) gave contradictory results. Only 1 sample was positive by LA test but not with ELISA or EM. The sensitivity of LA test and EM was 62.5 per cent (30 of 48) and 75 per cent (36 of 48); the corresponding specificity being 97.7 per cent (44 of 45) and 100 per cent (45 of 45) respectively. ELISA was more sensitive than the LA test and EM for detection of rotavirus antigen. LA test which is highly specific and a rapid method, may be useful in certain situations but its low sensitivity makes it unsuitable for use in routine clinical practice. PMID- 2550365 TI - A simple spot-test for circulating Entamoeba histolytica antigen-antibody complexes in patients with amoebic liver abscess. AB - An enzyme linked immunosorbent assay (ELISA) is described for the detection of E. histolytica immune complexes in serum. Antiamoebic antibody purified by affinity chromatography was used both to precoat strips of nitrocellulose membrane and as an enzyme carrier. These strips were incubated with samples of concentrated test serum and the enzyme conjugate. Following treatment with the peroxidase substrate 3-amino-9-ethylcarbazole the presence of E. histolytica antigens was visualized as red spots. Blocking of positive test sera with excess antibodies inhibited this reaction. Serum samples from 47 patients with amoebic liver abscess, 43 patients with other enteric diseases and 35 healthy controls were investigated. The spot test was positive in 75 per cent of patients with amoebic liver abscess, and in 12 per cent diseased controls whereas all the healthy controls were negative. The spot test is simple and sensitive and does not require prior separation of the immune complexes. The test is recommended as an aid to the diagnosis in patients suspected to have amoebic liver abscess. PMID- 2550366 TI - The sympathetic neuro-melanophore transmission in a fresh-water Indian major carp, Labeo rohita (Ham.). AB - Adrenaline was effective in aggregating the melanosomes both in innervated as well as denervated melanophores. Isotonic KCl could induce pigment aggregation only in innervated melanophores. Adrenaline- and K+-induced pigment aggregation response in these melanophores was blocked by phentolamine: propranolol failed to do so. It is suggested that the chromatic nerves in the fish, Labeo rohita are adrenergic and via post-synaptic alpha-adrenoceptors, control the melanosome aggregation. PMID- 2550369 TI - Molecular analysis of a region of the group B streptococcus chromosome involved in type III capsule expression. AB - Type III group B streptococci (GBS) are the most common cause of neonatal sepsis and meningitis in the United States. The important role of the type III polysaccharide capsule and of the terminal sialic acid moiety of the capsule in the virulence of GBS has been demonstrated by using Tn916 mutagenesis. Several of the transposon insertion sites that resulted in defective type III capsule synthesis were located in a 30-kilobase (kb) region of the chromosome. Hybridization analysis of two other type III strains that differed in their relative virulence and of GBS serotypes Ia, Ib, Ic, and II showed that this region of the chromosome was highly conserved. A repetitive 1.4-kb sequence was found only in the 30-kb region of the more virulent type III strain, COH 1. The Escherichia coli maxicell in vivo expression system and an in vitro coupled transcription-translation system successfully identified the proteins expressed from the 30-kb region. Comparison of the proteins expressed from the same DNA fragments in these two assays indicated that some of these proteins may contain leader sequences that would ultimately result in their secretion to the cell surface. Identification and further characterization of the genes and their products will provide the foundation for understanding the genetic and biochemical events in GBS capsular polysaccharide production. PMID- 2550367 TI - In vivo activation of peripheral blood polymorphonuclear neutrophils by gamma interferon results in enhanced fungal killing. AB - The effect of in vivo administration of murine recombinant gamma interferon (IFN) on the fungicidal activity of murine peripheral blood polymorphonuclear neutrophils (PB-PMNs) was studied. Mice were injected intramuscularly with 250, 2,500, 25,000 or 250,000 U of IFN 5 h before collection of peripheral blood. Purified PB-PMNs were cocultured in vitro with Blastomyces dermatitidis yeast cells for 2 h. PB-PMNs from untreated mice killed 44.5 +/- 12.5% of the fungal inoculum, whereas PB-PMNs from mice treated with 25,000 or 250,000 U of IFN showed significantly enhanced in vitro killing (68.0 +/- 9.4% [P less than 0.005] and 72.3 +/- 1.1% [P less than 0.001], respectively). Treatment with 250 or 2,500 U of IFN or 25,000 U of heated (100 degrees C, 15 min) IFN had no effect. The IFN induced activation of PB-PMNs was transitory. Significant enhancement of PB-PMN killing activity occurred 1, 2, or 5 h after in vivo IFN administration, but no enhancement was observed 16 or 24 h after IFN treatment. Enhanced fungicidal activity by PB-PMNs from mice treated for 5 h with 25,000 U of IFN correlated with an increased release of superoxide anion (O2-) in vitro after stimulation of PB-PMNs with phorbol ester; normal PB-PMNs and IFN-activated PB-PMNs, respectively, produced 2.2 +/- 2.5 and 23.5 +/- 4.8 nmol of O2- per 10(6) PB-PMNs per 30 min (P less than 0.005). The exogenous addition of compounds that antagonize or inhibit the formation of oxygen radicals (superoxide dismutase, catalase, dimethyl sulfoxide, or sodium azide) significantly inhibited fungal killing by both normal and IFN-activated PB-PMNs. In addition to the enhanced microbicidal activity and superoxide generation demonstrated in vitro with constant cell numbers, there was a transient leukocytosis (particularly neutrophilia) in peripheral blood at doses of IFN and at times after IFN administration where enhanced activity was also demonstrated. In summary, our results indicate that PB-PMNs can be activated in vivo for enhanced killing of a fungal target. The enhanced killing capacity of IFN-activated PB-PMNs is due at least in part to the enhancement of oxidative killing mechanisms. PMID- 2550368 TI - Adhesion to and invasion of HEp-2 cells by Campylobacter spp. AB - Twenty-one isolates were tested for their ability to adhere to and invade HEp-2 cells in vitro. Of the 21 organisms tested, 2 did not invade the HEp-2 cells, and 1 of these did not adhere to the epithelial cells. Campylobacter jejuni clinical isolates were more invasive than the nonclinical strains that were tested. When HEp-2 cells were treated with cytochalasin B, the invasiveness of C. jejuni was reduced, indicating active participation of the host cell in the uptake of these organisms. The number of intracellular C. jejuni isolates decreased when Campylobacter whole-cell lysates were absorbed onto HEp-2 cell monolayers. Experiments were also conducted to identify the functional sites of the antigens responsible for expression of Campylobacter invasion. Oxidation of lysates with sodium meta-periodate significantly affected its inhibitory capacity. This implies that the Campylobacter invasive ligand appears to be dependent upon an intact carbohydrate moiety. PMID- 2550370 TI - Modulation of leukotriene generation by pertussis toxin. AB - The purpose of our study was to characterize the properties of the interaction of pertussis toxin with human polymorphonuclear leukocytes for the modulation of leukotriene generation and metabolism. The cells were stimulated with either the Ca ionophore A23187, opsonized zymosan, or the bacterial peptide formyl-methionyl leucyl phenylalanine. Incubation of the cells with pertussis toxin led to a rapid inhibition of LTB4 generation when formyl-methionyl-leucyl phenylalanine was used as the stimulus, whereas there was no effect with the Ca ionophore and just a low effect with opsonized zymosan. The inhibition of leukotriene generation was dependent on the incubation time, temperature, and pertussis toxin concentration. The effect was not dependent on the presence of calcium. Incubation of the cells with guanosine 5'-O-(3-thiotriphosphate) the stable analog of GTP, led to a time dependent increase in leukotriene generation induced by formyl-methionyl-leucyl phenylalanine which was abolished by the simultaneous addition of pertussis toxin. Our data suggest that the formyl-methionyl-leucyl phenylalanine-induced generation of leukotrienes is dependent on a GTP-binding protein. The participation of the various G proteins has yet to be elucidated. PMID- 2550371 TI - Fusion of inclusions following superinfection of HeLa cells by two serovars of Chlamydia trachomatis. AB - We used a double-label immunofluorescence assay to examine the ability of Chlamydia trachomatis serovar F to infect and develop within HeLa 229 cells previously infected with serovar E. No exclusion to superinfection occurred for up to 24 h following infection by serovar E. The percentage of HeLa cells infected in cultures inoculated with both strains was identical to that of cells in cultures inoculated with one strain as a control. Organisms of both serovars were located within the same intracellular inclusion in 88 to 95% of HeLa cells infected with both serovars. The proportion of superinfected HeLa cells containing both strains in separate inclusions increased when there was exposure to inhibitors of cytoskeletal structure and transport. We used this inhibition to demonstrate that fusion of C. trachomatis phagosomes occurs throughout the developmental cycle. PMID- 2550372 TI - Sera of children with renal tumours contain low-molecular-mass hyaluronic acid. AB - The molecular mass of hyaluronic acid (HA) rather than its serum concentration alone may be a hallmark of certain types of malignancy. A radiometric assay was used to measure HA levels in 35 children with renal tumours [33 Wilms' tumours and 2 bone metastasizing renal tumours of childhood (BMRTC)] and 20 normal siblings of children with cancer. The HA level in the sera of normal children was barely detectable and had a molecular mass of 1-5 x 10(5). In both Wilms' and BMRTC patients, very high levels of HA were found in preoperative serum samples; these fell dramatically following surgical excision of the tumours. A novel finding of our study was the presence of low-molecular-mass HA (similar to the angiogenic fragments of HA) in the sera of BMRTC patients. In contrast, high molecular-mass HA (which is not angiogenic) was found in the sera of Wilms' patients (2 x 10(6) kDa). Following surgery in BMRTC patients, not only did serum HA levels fall to a value within normal ranges, but also the HA which remained was of high molecular mass. PMID- 2550374 TI - Oestrogen potentiates topoisomerase-II-mediated cytotoxicity in an activated subpopulation of human breast cancer cells: implications for cytotoxic drug resistance in solid tumours. AB - Primary resistance to chemotherapeutic agents is a major problem in the management of advanced cancer. By using oestrogen to modulate the topoisomerase II content of T-47D human breast cancer cells, we show here that cell subpopulations resistant to the topoisomerase-II-interactive drug VPI6 (etoposide) can be identified and quantified using single-cell analytical techniques. Immunohistochemical studies reveal topoisomerase II to be present in approximately 10% of control cells compared with 30% of oestrogen-stimulated cells, and this difference is reflected in the proportions of cells exhibiting VPI6-induced cell-cycle delay. This moderate increase in overall cell sensitivity is accompanied by massive enhancement of clonogenic cell kill, suggesting that oestrogen enhances VPI6 cytotoxicity by recruiting a clonogenic cell subpopulation characterized by increased topoisomerase II content. Flow cytometry confirms that the increase in topoisomerase II is localized to an activated G1 phase cell subset. We conclude that (i) single-cell analysis of cellular topoisomerase II content is predictive of VPI6 chemosensitivity; (ii) the existence of resistant tumour-cell subpopulations does not necessarily indicate the presence of phenotypically divergent subclones; and (iii) rational strategies for eliminating tumour resistance may be based on biological manipulation of specific cytotoxic drug targets. PMID- 2550373 TI - Transplantable Marek's disease lymphomas. III. Induction of MHC-restricted tumor immunity by lymphoblastoid cells in F1 hosts. AB - F1 chickens, which were a cross between birds of the related inbred lines G-B1 and G-B2, were immunized with in vitro cultured virus-non-producer lymphoblastoid cells that were either syngeneic or allogeneic with the challenge tumor cells. The lymphoblastoid cells were derived from Marek's disease herpesvirus (MDV) induced transplantable tumors. Previous findings showed that such immunization of G-B1 and G-B2 chickens prevented early mortality caused by the tumors. Lymphoblastoid cells syngeneic with the tumors were more effective than allogeneic cells, suggesting that an MHC-restricted immune response was induced, or, alternatively, that immune elimination of allogeneic cells prevented them from initiating strong immunity to MDV-associated tumor antigens. Immune elimination of the immunizing cells should not occur in F1 birds heterozygous for the 2 different parental MHC haplotypes (B6 and B13). Early mortality among F1 chickens immunized with lymphoblastoid cells that were syngeneic with the challenge tumor cells was significantly lower than for non-immunized control chickens or for birds immunized with lymphoblastoid cells that were allogeneic with the challenge tumor cells. Our results suggest that MDV-induced tumor antigens may be recognized by the host as altered-self MHC antigens. PMID- 2550375 TI - Integrin-receptor-mediated differentiation and growth inhibition are enhanced by transforming growth factor-beta in colorectal tumour cells grown in collagen gel. AB - We have studied the role of cell-matrix interactions and the modulating effect on these of transforming growth factor-beta s (TGF-beta s) in controlling differentiation and proliferation in a series of human colorectal carcinoma cell lines. Two (SW1222 and SW480) out of 7 cell lines specifically bound type-I collagen, via integrin-like (SW1222) and non-integrin-like (SW480) collagen receptors. Binding of these receptors may be responsible for regulating the degree of epithelial differentiation of the cells when grown in a 3-dimensional (3D) collagen gel. We have also shown that TGF-beta s enhance the binding of SW1222 cells to collagen and that this is accompanied by greatly increased crypt like glandular differentiation and inhibition of cell proliferation. Inhibition of cell proliferation was only seen when cells were grown in 3D collagen gel and were thus expressing a fully differentiated phenotype. The enhanced collagen binding induced by TGF-beta s was partially inhibited by an Arg-Gly-Asp (RGD) containing peptide which is a cell recognition signal for collagen binding. This suggests that TGF-beta s mediate their effects on differentiation of SW1222 cells specifically by modulating the expression of the integrin-like collagen receptor. The other colorectal carcinoma cell lines which lack this integrin-like receptor either failed to bind collagen or, in the case of SW480 binding, exhibited differentiation and proliferation which were not affected by TGF-beta s. This suggests that cell responsiveness to TGF-beta s may depend, at least in part, upon the cell-matrix interaction. PMID- 2550376 TI - Replication of latent Epstein-Barr virus genomes in normal and malignant lymphoid cells. AB - DNA replication of 2 human lymphoid cell lines (U296 and Raji), latently infected with the Epstein-Barr virus, has been compared using a density transfer approach. Typical of non-malignant lymphoblastoid cells, U296 cells divided once in bromodeoxyuridine-supplemented medium to form hybrid but not heavy-density host DNA. Replication of the intracellular Epstein-Barr virus DNA was selectively inhibited in these cells with only 15% of the viral genomes duplicating once to form hybrid-density viral DNA. However, some heavy-density viral DNA was formed in the U296 cells and DNA synthesis can thus initiate again on newly duplicated viral genomes in cells that have traversed only a single S phase. These results contrast strongly with observations concerning the Burkitt-lymphoma-derived cell line. Lymphoma cells are not growth-inhibited and most of the latent Epstein-Barr virus genomes of the Raji line replicated once, and only once, in successive S phases. While the majority of the 50 Epstein-Barr virus genomes of both the Raji and U296 cell lines are maintained as extra-chromosomal DNA plasmids, the control of their duplication is distinctly different in the respective malignant and non malignant host cells. PMID- 2550377 TI - Long-lasting signals in mitogenesis and transformation. PMID- 2550378 TI - Angiotensin-converting enzyme as a possible marker for lung toxicity in amiodarone-treated patients. AB - Since it has been observed that in vitro amiodarone induces morphological alterations in endothelial cells similar to those observed in patients with lung toxicity and that the angiotensin-converting enzyme (ACE) seems to be a marker for perturbation of the alveolar-capillary membrane, serum ACE concentrations have been determined in 44 patients, 23 treated with amiodarone (group A) and 21 treated with other anti-arrhythmic drugs (group B), before the beginning of treatment and after 7, 15, 30, 60 and 180 days. Serum ACE concentrations in group A were lower than the basal values (15.8 +/- 5.9 mU/ml) on day 7 (12.7 +/- 4.5 mU/ml) and were higher on day 60 (17.9 +/- 3.8 mU/ml), then returned to basal values by day 180 (15.9 +/- 5.5 mU/ml), but none of the differences were statistically significant. In group B, serum ACE concentrations were significantly higher than basal values (15.2 +/- 4.0 versus 14.2 +/- 3.5 mU/ml, p less than 0.05) only on day 15. In group A serum ACE concentrations were significantly higher than in group B only on day 60 (17.9 +/- 3.8 versus 14.7 +/- 4.5 mU/ml, p less than 0.025). During the period of the study none of the patients showed any clinical or radiological signs of lung toxicity or reduction of lung diffusion capacity for carbon monoxide (DLCO). Serum ACE levels were normal even in three patients who developed pulmonary fibrosis and in four whose DLCO was reduced by more than 20% from the basal values after the study was completed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550379 TI - Pemphigus foliaceus associated with acanthosis nigricans-like lesions and hepatocellular carcinoma. PMID- 2550380 TI - Synthesis, characterization and inhibitory activities of (4-N3[3,5-3H]Phe10)PKI(6 22)amide and its precursors: photoaffinity labeling peptides for the active site of cyclic AMP-dependent protein kinase. AB - PKI(6-22)amide is a 17 residue peptide corresponding to the active portion of the heat-stable inhibitor of cAMP-dependent protein kinase. The peptide is a potent (Ki = 1.6 nM), competitive inhibitor of the enzyme. The photoreactive peptide analog (4-azidophenylalanine10)PKI(6-22)amide was synthesized in both its non radiolabeled and tritiated forms by chemical modification of precursor peptides that were prepared by stepwise solid-phase synthesis. (4-Amino[3,5 3H]phenylalanine10)PKI(6-22)amide, the precursor for the radiolabeled arylazide peptide, was obtained by catalytic reduction of the corresponding peptide containing the 3,5-diiodo-4-aminophenylalanine residue at position 10. The purified PKI peptides were analyzed by HPLC, amino acid analysis, and u.v. spectra. In the dark, (4-azidophenylalanine10)PKI(6-22)amide inhibited the catalytic subunit of cAMP-dependent protein kinase with a Ki value of 2.8 nM. The photoreactivity of the arylazide peptide was demonstrated by time-dependent u.v. spectral changes on exposure to light. Photolysis of the catalytic subunit (4 azido[3,5-3H]phenylalanine10)PKI(6-22)amide complex resulted in specific covalent labeling of the enzyme. The data indicate that this peptide is a useful photoaffinity labeling reagent for the active site of the protein kinase. PMID- 2550381 TI - Intrinsic effects of the benzodiazepine receptor antagonist RO 15-1788 in sleepy and alert subjects. AB - The present paper investigated the differential effects of 5 mg, 60 mg and 120 mg of the benzodiazepine receptor antagonist RO 15-1788 on the ability to resist sleep of sleepy and alert subjects. Repeated administrations of 5 mg potentiated the hypnotic effects of sleep-deprivation in alert subjects and decreased the hypnotic effects in sleepy subjects. Similar differential effects were found with respect to two factors of subjective mood. The two higher levels of the drug decreased sleepiness more in sleepy than in alert subjects. These results are interpreted to support the hypothesis that the effects of RO 15-1788 may be dependent on the level of an endogenous benzodiazepine-like sleep factor. PMID- 2550382 TI - Influence of eicosapentaenoic acid and vitamin E on brain cortex Ca2+ ATPase activity in cholesterol-fed rabbits. AB - The influence of eicosapentaenoic acid (EPA) and vitamin E on brain cortex Ca2+ ATPase activity was examined in rabbits receiving cholesterol-rich diets for a period of 45 days. Rabbits were divided as control (A) and cholesterol-fed groups (B, C, and D). Group C received 80 mg of EPA and group D received 100 IU of vitamin E every day in addition to the cholesterol-rich (2%, w/w) diet which was solely given to Group B. Rabbits receiving cholesterol alone had a significant reduction in brain microsomal phospholipid level. Microsomal free cholesterol and polyunsaturated fatty acids (PUFA) were significantly increased in all experimental groups. Cortex microsomal Ca2+ ATPase activity was found to be inhibited in all cholesterol-fed rabbits as compared to controls, but the highest inhibition was seen in rabbits fed cholesterol alone. Additions of EPA or Vitamin E to the cholesterol-rich diets resulted in a recovery of the enzymatic activity. It is concluded that cholesterol feeding without any addition of PUFA or antioxidant agent might cause an inhibition of brain Ca2+ ATPase activity in rabbits, thereby leading to the dysfunction in ion transport and neurotransmitter release. PMID- 2550383 TI - Quantification of plasma cortisol levels in ascorbic acid studies using guinea pigs implanted with ACTH pellets. AB - A method is described that permits multiple blood collections from unanesthetized, unrestrained guinea pigs with minimal disruption to the animal, as verified by plasma cortisol (PC) levels. Blood (100-200 microliters) was collected from the lateral metatarsal vein using a vacuum-assisted method. Using this method, the effect of subcutaneously-implanted slow-releasing ACTH versus placebo pellets on PC levels was determined. A small, but significant, increase in PC concentration occurred in the control group, reflecting the stress of handling and the implant procedure. In contrast, the peak concentration of PC after implantation of a pellet containing ACTH was 20 times the starting level and occurred within 4 hr. However, the PC level was not correlated with the content of ACTH in the pellet because a similar level was achieved with 0.125 or 8 mg/pellet. A decline in Pc to pre-treatment levels occurred within 24 to 48 hr. The results indicate that administration of ACTH by pellet, although intended to achieve a sustained elevation in PC level, is not suitable for maintaining elevated PC levels. PMID- 2550385 TI - Tumor volume and treatment outcome in small cell lung carcinoma. AB - The volume of some tumors correlates with local disease control, response, and survival. Small cell lung carcinoma presents with variable and often large intrathoracic masses. We assessed volume by two methods (summation of cross sectional areas and Simpson's rule for estimating volume) and correlated volume with treatment outcome. Twenty-nine patients with small cell lung carcinoma (12 limited disease and 17 extensive disease) were analyzed for volume outcome correlations. There were five early deaths. Twenty-four had response correlations (11 limited disease and 13 extensive disease). Eighteen of the twenty-four achieved complete response. There was no apparent correlation between complete and/or partial response and pretreatment tumor volume. There was no apparent correlation between survival and volume. The methods of volume assessment correlate well with each other (r = 0.88) and either can be used with accuracy. So far there is also no correlation between chest tumor volume and intrathoracic recurrence. Our projections suggest that there is a low probability of correlating chest tumor volume and either response or survival. This is in keeping with the disseminated nature of the tumor which is its limiting factor in survival. PMID- 2550384 TI - Increased axonal transport of trigeminal ganglion proteins in an electrode model. AB - This investigation uses an electrical stimulation model of ocular herpes simplex virus (HSV) reactivation to elucidate the relationship between neuronal protein synthesis, axonal transport and electrical stimulation. In this study, healthy adult albino rabbits were implanted with electrodes over the trigeminal ganglion (TG) and stimulated chronically twice a week for 28-34 days (previously shown to induce HSV reactivation in infected rabbits). The TG was visualized surgically and injected with 3H-proline to label newly synthesized protein. After various times, the TG, corneas and ophthalmic nerve were analyzed for labeled proteins. In nonimplanted and implanted rabbits (whether stimulated or not), the rate of fast anterograde axonal transport in the ophthalmic nerve was 347 mm/day and 326 mm/day, respectively. A lag period of 45 min occurred between initiation of protein labeling and onset of axonal transport. Only a small percentage of newly synthesized neuronal proteins were transported in the ophthalmic nerve (1-4% in the proximal 10 mm) or deposited in the cornea (less than 0.5%) after 24 hr. Neither the rate of protein synthesis in the TG nor the rate of axonal transport was changed by electrical stimulation. However, the amount of radioactively labeled proteins undergoing axonal transport in the stimulated group was five times that of the sham control group. Thus, the routing and loading of TG proteins for axonal transport is enhanced in the electrical stimulation model. PMID- 2550386 TI - Amphibian and piscine iridoviruses proposal for nomenclature and taxonomy based on molecular and biological properties. AB - We have compared a number of properties of the well-characterized iridovirus, frog virus 3, with two other iridoviruses from amphibia, bullfrog edema virus and Lucke triturus virus, and with a piscine iridovirus, goldfish virus (GFV), to provide information for developing taxonomic classification of these viruses and establishing their ecological niche. Purified virions had similar size and shape (icosahedral) for each virus, and the genomic DNAs of each virus were methylated by a virus-induced DNA methyltransferase. The three amphibian viruses replicated equally well in fish (FHM), hamster (BHK), and human (WI-38) cell monolayer with identical cytopathology, while GFV failed to replicate in these cell lines. However, GFV replicated albeit at a slow rate, in a goldfish cell line; there was no detectable replication by the amphibian viruses in these cells. The amphibian iridoviruses had virtually similar DNA sequences, while those of GFV were markedly different. Analyses of virus-induced polypeptides in infected cells corroborated the DNA analyses; the polypeptides of the amphibian viruses were similar and distinct from those of the fish virus. Nongenetic reactivation could only be accomplished between the three amphibian viruses but not with the piscine virus. Based on these data, we suggest taxonomic and nomenclature designations of amphibian and piscine iridoviruses. PMID- 2550387 TI - Enteroviral-related antigen in circulating immune complexes of amyotrophic lateral sclerosis patients. AB - Circulating immune complexes were isolated by polyethylene glycol precipitation from the sera of patients with amyotrophic lateral sclerosis. Rabbits immunized with circulating immune complexes from 3 of 5 amyotrophic lateral sclerosis patients induced antisera that specifically reacted with enterovirus-infected cells by immunofluorescence and enzyme-linked immunosorbent assay. These antisera were nonneutralizing and did not react with purified virus. In addition, peripheral lymphocytes of amyotrophic lateral sclerosis patients produced lymphokine in response to extracts from enterovirus (Coxsackie B4) infected cells. These results suggest both a humoral (circulating immune complex) and a cellular immune response in some patients with amyotrophic lateral sclerosis to enterovirus-coded or -induced antigen. PMID- 2550389 TI - Genome typing of three serotypes of porcine adenovirus. AB - Porcine enteric adenovirus serotypes 1-3, previously shown to be distinct antigenically, were purified by density gradient centrifugation, and the DNA was extracted. Fragments generated by digestion with restriction endonucleases BamHI, EcoRI, HindIII, and PaeR7 were separated by electrophoresis and compared with human adenovirus type 2 as a control. The results demonstrated that the three porcine viruses were closely related to each other, although distinguishable, and distinct by restriction endonuclease analysis from human, murine, canine, bovine, and fowl adenovirus. PMID- 2550388 TI - Expression of interleukin 2 receptors in T cells transformed by strains of Herpesvirus saimiri representing three DNA subgroups. AB - Eight T lymphoblastoid cell lines transformed by Herpesvirus saimiri were studied for requirement of interleukin 2 (IL-2) for growth in tissue culture and expression of the IL-2 receptor. The results show that four cell lines grew independent of IL-2, but four other cell lines required IL-2 for optimal growth. However, all H. saimiri transformed cells expressed IL-2 receptors, regardless of IL-2 requirement. Between 2,250 and 8,800 high-affinity receptor sites/cell were detected by ligand-binding assays; in contrast, only about 1,000 sites/cell were in an uninfected IL-2-dependent marmoset T cell line. Surface-bound IL-2 was internalized and antibodies to the receptor inhibited cell growth. Expression of IL-2 receptors may allow expansion of tumor cells in vivo in response to endogenous or exogenous IL-2. PMID- 2550390 TI - Radionuclide contents in building materials used in Hong Kong. AB - The radionuclide contents of conventional natural raw building materials, coal ash and slag, and finished building products have been determined using gamma-ray spectrometry. Results of brick measurements in their original geometry and in crushed form are compared. The radioactive concentrations in cement and sand, mostly imported from China, are among the lowest measured. However, due to the high radioactivity of aggregates, composed of granite mainly extracted locally, the mean Ra equivalent activity of concrete is high compared with that in some countries. The radioactivity levels of coal ash and slag in Hong Kong are about the average values in other countries. The incorporation of coal ash and slag in ordinary concrete does not alter the radioactivity significantly. PMID- 2550391 TI - Correlation of lung dose with Rn concentration, potential alpha-energy concentration and daughter surface deposition: a Monte Carlo analysis. AB - In the evaluation of lung cancer risk from Rn daughters in the home environment, measurements of Rn concentration or potential alpha-energy concentration (PAEC) have served as proxies for actual radiation dose to the lungs. This paper uses a single-compartment room model, model parameters from a number of recent studies, and Monte Carlo analysis to explore the relative value of Rn concentration, PAEC, and room surface deposition, or plate-out, as estimators of lung dose. Results indicate that Rn concentration and PAEC are fairly good estimators, explaining roughly 50-70% of the variation in dose for the conditions studied, over the range of Rn concentrations considered (74-555 Bq m 3). The relative advantage of one measure over the other depends upon the variability in a number of factors across the houses being evaluated. Room surface activity deposition was not found to be superior to the traditional measures of Rn concentration and PAEC. Significant room for improvement remains in the development of simple home monitors providing an improved estimation of lung dose. PMID- 2550392 TI - Synthesis of a high specific activity 125I-labeled analog of PK 11195, potential agent for SPECT imaging of the peripheral benzodiazepine binding site. AB - The peripheral benzodiazepine binding site ligand PK 11195 has been 125I-labeled by direct displacement of aromatic chlorine under solid-state conditions in 50 76% radiochemical yield and greater than 94% radiochemical purity. Purification by high pressure liquid chromatography increased the specific activity of the product from an initial 15-17 Ci/mmol to a final activity of 260-910 Ci/mmol. To determine the affinity of this [125I]PK 11195 analog for human glioma cells, saturation experiments were performed on monolayers of U251 human glioblastoma cells. Scatchard analysis of saturation data demonstrated that the [125I]PK 11195 analog binds to a single class of sites with a KD of 8.0 +/- 1.7 nM and maximal binding of 3.8 +/- 0.1 pmol/mg protein. These values are similar to those obtained when [3H]PK 11195 was assayed in U251 cells (KD = 14 +/- 3.4, Bmax = 4.1 +/- 1.3) suggesting that iodination does not appreciably alter the binding of PK 11195 to human glioma cells. In vivo autoradiographic studies of brain in C6 glioma bearing rats demonstrate selective binding of the radioligand to the tumor. These results suggest that this [125I]PK 11195 analog may be a useful radiotracer for the study of peripheral benzodiazepine binding sites. PMID- 2550393 TI - [Colostomy patients--well cared for, if counseled. Experiences of a colostomy counselling center]. AB - The postoperative care of stoma patients concerns the surgeon, the nurse, the relatives as well as the family doctor. In past years, specialized stomatherapy services became more important, and they represent a real necessity for the patients, according to our own experience. We plead for a broad stoma patient care including stomatherapy services in all cases, either during hospitalization or as an out-patient service after having left hospital. PMID- 2550394 TI - The immunocytochemical localization of superoxide dismutase in the enterocytes of the avian intestine: the effect of vitamin D3. AB - Both light microscopical and electron microscopical immunocytochemical techniques were utilized to localize CuZn-superoxide dismutase (SOD) in the duodenum of normal, rachitic and vitamin-D3-replete chicks. This enzyme catalyses the dismutation of the superoxide anion, a toxic free radical generated during the normal aerobic metabolism of most respiring cells. Light microscopy showed no SOD activity associated with the duodenal enterocytes of normal and rachitic chicks. However, in rachitic animals subsequently treated with vitamin D, i.e. vitamin-D replete chicks, intense immunoreactivity for the enzyme was seen in association with the apical border of the duodenal absorptive cells. Immunostaining for SOD was not seen in goblet cells. With electron microscopy, immunostaining for SOD activity was identified in association with the apical microvilli and, to a lesser degree, with the terminal web, a well as in association with both lysosomes and peroxisomes. From this report it appears that there is a physiological relationship between vitamin D, SOD and the intestinal absorptive cell. However, the precise relationship must await further clarification. PMID- 2550395 TI - Conservative treatment of uveal melanoma: local recurrence after proton beam therapy. AB - Twenty-three of 1006 (2.3%) uveal melanoma patients treated with proton beam therapy at the Harvard Cyclotron Laboratory between July 1975 and December 31, 1986 received additional treatment for documented (15 patients) or suspected (eight patients) tumor growth in the irradiated eye. Growth within the initially irradiated volume was documented at Massachusetts Eye and Ear Infirmary in 12 patients. Documented growth occurred in nine of 665 (1.4%) patients with small and intermediate size tumors, at times after treatment ranging from 6 to 48 months (median 16 months), and in three of 341 (.9%) patients with large tumors at 7, 11, and 12 months after treatment. Melanoma growing totally outside the treated volume was also documented in three additional patients at 7, 9, and 45 months; two of these were thought to be "ring melanomas". Eight patients had the treated eye removed elsewhere for suspected tumor growth. The additional treatment in these 23 patients was conservative in nine patients (repeat proton irradiation in five and laser photocoagulation in four). Thirteen underwent immediate enucleation and one had orbital exenteration. Ultimately, 17 of the 23 eyes (74%) were removed. Estimated probability of local control of the melanoma within the irradiated eye at 60 months was 96.3 +/- 1.5%. Dose distributions to the 12 patients with documented local failure within the irradiated volume were analyzed. Ten tumors recurred marginally in an area receiving less than the prescribed dose of 70 CGE (CGE = Cobalt Gray Equivalents = proton Gy X RBE 1.1), whereas only two recurred in the volume receiving full dose. Based on these data, it appears that a dose of 70 CGE in five fractions is associated with very high rates of local control in human uveal melanoma. It is reasonable to consider initiating studies using a lower total dose or a more protracted course, to determine if some of the observed complications are dose-related. PMID- 2550396 TI - Treatment results among adults with childhood tumors: a 20-year experience. AB - Controversy exists regarding the most appropriate treatment for the rare adult patient who develops a so-called pediatric cancer. We have reviewed our 20-year experience with these patients and analyzed their outcome. A total of 299 patients with rhabdomyosarcoma (106), Wilms' tumor (97), and neuroblastoma (96) were evaluated and treated at Stanford University Medical Center between January 1967 and December 1987. Only 26 of these patients (8.7%) were diagnosed during "adulthood"; their age range was 18-67 years, median 23 years. Wilms' tumor; Five patients presented with Wilms' tumor at age greater than or equal to 18 years; four had unfavorable histology. All underwent multimodality therapy; however, only two have survived, one currently disease-free and one with disease. Neuroblastoma: Five patients presented with neuroblastoma at age greater than or equal to 18 years. Four underwent attempted surgical resection, post-operative irradiation (RT), and chemotherapy (CT); the other received no adjuvant CT. Only two of the five patients survive, both with disease. Rhabdomyosarcoma: Of the 16 adults (greater than or equal to 21 years) with rhabdomyosarcoma, 14 (87%) had advanced Intergroup Rhabdomyosarcoma Study-group disease (eight Group III, six Group IV). All 16 underwent aggressive multimodality therapy. At 10 months-16 years follow-up, only five patients survive, four of whom are apparently cured of their tumor. Neither histologic subtype nor site of presentation were of prognostic value. This series demonstrates that adults with Wilms' tumor, neuroblastoma, or rhabdomyosarcoma have a worse prognosis than do children with the same diagnosis. Possible explanations for this disparity in outcome include different tumor biology, less tolerance for treatment, and different natural history among adults relative to children. PMID- 2550397 TI - Peripheral neuropathies following experimental intraoperative radiation therapy (IORT). AB - Injury to peripheral nerves in the lumbar para-aortic region was evaluated in beagle dogs 2 years following fractionated irradiation (EBRT), intraoperative irradiation (IORT), or a combination of IORT and EBRT. Time to onset of peripheral neuropathy was determined by means of serially completed neurological and electrophysiological examinations. Peripheral neuropathies were seen beginning as early as 6 months following 35 Gy (or greater) IORT only and 35 Gy plus 50 Gy EBRT. The incidence of peripheral neuropathies increased with increasing IORT doses beginning at 15 Gy. Onsets of peripheral neuropathies following IORT alone were clustered between 6 and 18 months, with onset in some dogs occurring as late as 24 months. The combination of IORT and EBRT resulted in an incidence and latency to onset of neuropathies similar to that seen with IORT alone. Neuropathies were not seen with EBRT alone at doses from 50 Gy to 80 Gy. Recovery of nerve function did not occur in affected dogs. Histological studies of nerves 2 years following irradiation demonstrated loss of axons and myelin, with a corresponding increase in endoneurial, perineurial, and epineurial connective tissue. Percentage of axon and myelin decreased to about 60% of normal at 15 Gy IORT, and additionally at higher doses. An insignificant decrease in percentage of axon and myelin was seen following EBRT alone. A significant lesion occurring in and around nerves at most IORT doses was necrosis and hyalinization of the media of small arteries and arterioles. The dose for a 50% probability for causing severe vessel lesions in the 2-year study was 19.5 Gy IORT only and 18.7 Gy when IORT was combined with EBRT. These lesions were not seen with any EBRT only dose. These studies suggest that peripheral nerve is a dose limiting normal tissue in IORT. Neuropathies appear to result from direct effects of irradiation on nerve and secondary effects to nerve resulting from damage to regional vasculature. PMID- 2550398 TI - Frequency, sites of relapse, and outcome of regional node failures following conservative surgery and radiation for early breast cancer. AB - Between 1970 and 1986, 990 patients underwent excisional biopsy and radiation for clinical Stage I or II breast cancer. A limited axillary dissection (levels I and II) was performed in 914 of these patients. The median follow-up was 40 months from the initiation of radiation. Thirty-one patients developed a regional node failure as their first site of recurrence either with (12 patients) or without (19 patients) simultaneous distant metastases. The median interval to recurrence was 27 months (range 4-59). The 5-year actuarial rate for an isolated regional node recurrence (without simultaneous distant metastases) was 3%. The most common site for a regional node failure was the axilla (17 patients) followed by the supraclavicular nodes (13 patients). Salvage therapy was effective for an axillary +/- breast failure with 10/14 patients alive with no evidence of disease. Prognosis was related to the site of recurrence as well as the presence or absence of distant metastases. The 5-year actuarial survival from initial treatment for all patients with a regional node failure was 63% with a 3-year actuarial survival of 57% from diagnosis of recurrence. Regional node failure was related to the number of axillary nodes removed at the time of dissection and patient age. PMID- 2550399 TI - The predictors of distant relapse following conservative surgery and radiotherapy for early breast cancer are similar to those following mastectomy. AB - Although previous studies have indicated that the predictors of local recurrence following conservative surgery (CS) and radiotherapy (RT) are not the same as those following mastectomy, it remains unclear whether the predictors of distant relapse differ by local treatment modality. Clinical and pathologic features predictive of distant relapse for patients treated with mastectomy have been well established and include lymph node involvement, histologic grade, and peritumoral lymphatic vessel invasion (LVI). To study the influence of these and other factors on the rate of distant relapse in patients treated with CS and RT, we have identified a group of 438 patients treated between 1968 and 1981 who met the following criteria: primary tumor size less than or equal to 5 cm, excision of the primary tumor, infiltrating ductal carcinoma as the most aggressive histologic subtype, histology evaluable for the presence of an extensive intraductal component, and a dose to the primary site greater than or equal to 60 Gy. Estrogen receptor status was available in 58% of cases, 76% had an axillary dissection, and 23% were treated with adjuvant chemotherapy. With a median follow up of 89 months, 107 patients (24%) developed a distant relapse. The 5-year actuarial freedom from distant relapse (FDR) was 80%. Stepdown Cox proportional hazards regression analysis identified several factors associated with a significantly (p less than 0.01) increased risk for distant relapse: positive lymph nodes, histologic grade, necrosis, and lymphatic vessel invasion. The magnitude of each effect was then examined with a lifetable calculation. Five year freedom from distant relapse was 86% for the node-negative subgroup, 78% for patients with one to three positive nodes, and 45% for patients with four or more positive nodes. For histologic grades I, II, and III, 5-year freedom from distant relapse was 96%, 97%, and 75%, respectively. For necrosis scored as absent, scant, moderate, or marked, 5-year freedom from distant relapse was 90%, 78%, 77%, and 66%, respectively. For lymphatic vessel invasion scored as absent or present, 5-year freedom from distant relapse was 85% and 63%, respectively. We conclude that the clinico-pathologic predictors for distant relapse following conservative surgery and radiotherapy appear to be the same as those following mastectomy. This observation is consistent with the notion that distant relapse is caused by the presence of micrometastases at the time of initial patient sentation and is not greatly influenced by selection of local treatment. PMID- 2550401 TI - Protection of bone marrow by WR-2721 after fractionated irradiation. PMID- 2550400 TI - Primary malignant mediastinal germ cell tumors: a study of eleven cases and a review of the literature. AB - From 1957 to 1988 eleven cases of primary mediastinal germ cell tumor were referred to the Peter MacCallum Cancer Institute (PMCI). Four were seminomas, three were mixed germ cell tumors, two were embryonal carcinomas and two were teratocarcinomas. Two of the eleven patients were female. For seminoma, surgical debulking and post-operative irradiation produced the best results. Mediastinal doses ranged from 30 to 37.5 Gy. Local control was achieved in all cases; two patients survive disease-free. The non-seminomatous germ cell tumors showed a significantly poorer survival with only one of seven patients remaining alive in remission at 15 months. One other case of non-seminomatous tumor remains alive but in relapse at 23 months. Attention is focused on the anterior position of primary germ cell tumors in the mediastinum. A review of the literature is presented. PMID- 2550402 TI - Effect of MK-906, a specific 5 alpha-reductase inhibitor, on serum androgens and androgen conjugates in normal men. AB - To determine the hormonal effects of MK-906, an orally active 5 alpha-reductase inhibitor, on serum androgens and androgen conjugates, 12 healthy men were given 10, 20, 50, and 100 mg MK-906 2 weeks apart in randomized order in a 4-period crossover design. Serum testosterone (T), dihydrotestosterone (DHT), androstanediol glucuronide, and androsterone glucuronide were measured before and 24 hours after each dose. The effect of MK-906 on glucuronyl transferase activity, the enzyme responsible for androstanediol glucuronide and androsterone glucuronide formation, was assessed in vitro using rat prostate tissue. Serum T levels were unchanged after all doses. Serum DHT, androstanediol glucuronide, and androsterone glucuronide were suppressed by 70, 40, and 56%, respectively, after the 10-mg dose, and by 82, 52, and 66% after the 100-mg dose (P less than 0.02 for the comparison between the 10 and 100-mg doses for all three steroids), respectively. Baseline serum T and DHT levels were strongly correlated (R = 0.89, P = 0.0002), as were androstanediol glucuronide and androsterone glucuronide levels (R = 0.78, P = 0.003), but there was no correlation between DHT levels and the levels of either conjugated steroid. MK-906 had no effect on glucuronyl transferase activity in vitro. It was concluded that single doses of MK-906 suppress both conjugated and unconjugated 5 alpha-reduced androgens. While all three steroids appeared to be good markers of systemic 5 alpha-reductase inhibition, further research will be needed to determine which steroid best reflects tissue DHT levels in patients receiving these inhibitors. PMID- 2550403 TI - The actions of calcitonin on the TM3 Leydig cell line and on rat Leydig cell enriched cultures. AB - Studies demonstrating calcitonin receptors on Leydig cells have suggested that these cells may be one of the many sites affected by this peptide. To investigate this possibility, the effect of synthetic salmon calcitonin on the TM3 Leydig cell line (derived from immature mouse Leydig cells) and on primary Leydig cell enriched preparations was examined. Synthetic salmon calcitonin stimulated the conversion of [3H]adenine to [3H]cyclic AMP in TM3 cells. In addition, the hormone stimulated the basal secretion of testosterone in both TM3 cell- and Leydig cell-enriched cultures and potentiated the action of hCG on Leydig cell enriched cultures. Synthetic salmon calcitonin also increased the concentration of androgen and estrogen receptors in cultured TM3 Leydig cells by 2- and 4-fold, respectively, when added to the culture medium (1 micrograms/ml). The fact that 8 bromo-cyclic AMP decreased both androgen and estrogen receptor concentrations suggested that the effect of calcitonin on sex steroid receptors is not mediated by its effect on cyclic AMP in these cells. The possibility that the action of calcitonin on steroid receptors might be mediated by another messenger such as calcium (Ca2+) was therefore considered. Progressively lowering the concentration of Ca2+ in the culture medium of the cells from 1.5 mM to less than 0.01 mM decreased the concentration of both androgen and estrogen receptors. Returning the Ca2+ concentration to normal levels (1.5 mM) restored steroid receptor levels. Receptor levels were also decreased when the extracellular Ca2+ concentration was lowered to 0.5 mM, and treatment with the Ca2+ ionophore, A23187 (1 microM), restored receptor levels to normal. The calcium channel blocker, verapamil, decreased the androgen receptor concentration but unexpectedly increased the concentration of estrogen receptors. It was concluded that calcitonin stimulates cAMP formation and testosterone secretion, and increases the concentration of sex steroid receptors. These observations provide evidence that the previously demonstrated calcitonin receptors on Leydig cells may be coupled to several biologic responses in this cell type. PMID- 2550404 TI - The action of calcitonin on the TM4 Sertoli cell line and on rat Sertoli cell enriched cultures. AB - The effects of synthetic salmon calcitonin on primary Sertoli cell-enriched cultures and on an established cell line (TM4 cells, derived from immature mouse Sertoli cells) were studied. Synthetic salmon calcitonin stimulated the conversion of [3H]adenine to [3H]cyclic AMP in both cell systems. In addition, this peptide stimulated the secretion of rABP in primary Sertoli cell-enriched cultures prepared from rat testis. Calcitonin also increased the total concentration of both androgen and estrogen receptors in TM4 cells. Because cAMP analogs decreased androgen and estrogen receptor concentrations, the effect of calcitonin on sex steroid receptors may not be mediated by its effect on cyclic AMP in these cells. The possibility that the action of synthetic salmon calcitonin on the receptors might be mediated by a change in cellular Ca2+ was investigated. Lowering extracellular Ca2+ concentrations from 1.5 mM to less than 0.01 mM markedly reduced the concentration of androgen and estrogen receptors; restoration of Ca2+ to 1.5 mM returned receptor levels to normal. When the receptor concentrations were decreased by lowering extracellular Ca2+ concentrations to 0.5 mM, treatment with the calcium ionophore, A23187, restored receptor levels to normal. Although the calcium channel blocker, verapamil, decreased receptor levels, calcitonin partially counteracted its effect. Trifluoperazine, an inhibitor of calmodulin, also diminished androgen and estrogen receptor, levels in the cytosol of TM4 cells. It was concluded that calcitonin stimulates the formation of cyclic AMP and the secretion of rABP by Sertoli cells. This peptide also increases the concentration of androgen and estrogen receptors, possibly by a mechanism that is, in part, Ca2+ -mediated. These results, along with those on Leydig cells, suggest that calcitonin could be a regulator of testicular function. PMID- 2550406 TI - Cytomegalovirus: the disease and its management. Based on a meeting. London, 11 October 1988. PMID- 2550405 TI - The spectrum of cytomegalovirus infection and its management. AB - Serious disease due to cytomegalovirus (CMV) is almost totally confined to congenitally infected infants and the immunocompromised. Cell-mediated immunity is primarily responsible for the control of infection following primary infection and for prevention of disease after reactivation. In patients receiving active immunosuppression for transplantation, the choice of drug regimens is important in determining the likelihood of disseminated CMV infection and disease and it may be possible to control the infection by reducing the doses used. A number of strategies have been introduced in attempts to reduce the morbidity and mortality of CMV in the immunocompromised. These include prophylactic administration of specific immunoglobulin or acyclovir and the therapeutic use of several antiviral agents shown to have activity against CMV in vitro. Ganciclovir (DHPG) and foscarnet (phosphonoformate) have shown the most promise in clinical trials and ganciclovir has now been licensed for use in the UK, in sight- and life threatening infections. PMID- 2550407 TI - Diagnosis of cytomegalovirus infection. AB - CMV is an important pathogen for immunocompromised patients. Since such individuals cannot be relied upon to mount appropriate immune responses promptly, methods for detecting virus must be used for diagnosis. Conventional virus isolation is slow and the sensitivity and specificity of a more rapid method using monoclonal antibodies are described. Other potential ways of detecting cytomegalovirus are discussed, including the difficulties of detecting this virus immunochemically in clinical specimens due to its coating of human beta 2 microglobulin. PMID- 2550408 TI - Cytomegalovirus in the neonate. AB - Cytomegalovirus infection in early life may be congenital, or acquired during delivery or in the post natal period. The incidence of congenital infection varies widely throughout the world and ranges from 0.2 to 2.2% of live births; rates in the United Kingdom are between 0.3 and 0.4%. While fewer than 10% of congenitally infected infants have clinical manifestations of CMV infection at birth, most of this group will have serious neurological handicap. Of those with no symptoms at birth about 5% will subsequently manifest CMV-related problems, the most common being sensorineural hearing loss. Most congenitally infected children are neurologically and intellectually normal. Infection is acquired early in a high proportion of infants. In one study where 50% of mothers had CMV antibodies, 12% of their infants had acquired infection by three months and 20% by one year. The mothers' gestational status and whether the infants were breast fed were major factors associated with infection. Transfusion of CMV-infected blood is another important source of CMV infection and screened blood should be given to premature infants. PMID- 2550409 TI - Prophylaxis and treatment of cytomegalovirus infections in the bone marrow transplant recipient. AB - Almost all viral infections after bone marrow transplant (BMT) are attributable to reactivation of latent DNA viruses, especially those of the Herpesvirus family. The first to reactivate is herpes simplex virus, followed by cytomegalovirus (CMV) with a reported peak incidence between 60 and 90 days in most studies and varicella zoster virus about three to 18 months after BMT. Epstein-Barr virus rarely causes detectable illness after BMT and reactivation is only seen in the most profoundly immunosuppressed. The most lethal infection and indeed the most prominent cause of failure overall after allogenic BMT is interstitial pneumonitis which is frequently due to CMV infection. This article considers prophylactic measures against viral infection and preliminary studies of regimens for the treatment of established CMV infections. PMID- 2550410 TI - Cytomegalovirus infection in the acquired immune deficiency syndrome. AB - CMV disease is a major problem in AIDS, though with a different profile from that seen in other immunosuppressed patients. The novel treatments, ganciclovir and phosphonoformate, have a major role in the management of such infections, but maintenance therapy is often required. Optimal maintenance regimens have yet to be established, especially where zidovudine is also being used. PMID- 2550411 TI - Cytomegalovirus infection and renal transplantation. AB - Cytomegalovirus (CMV) infections remain a major clinical problem after renal transplantation. Reported incidences of CMV disease range from 17 to 25% in patients with azathioprine treatment and from 2 to 23% in patients on cyclosporin A. CMV-related death occurs in 1-3% of the total kidney transplant population. CMV seronegative kidney recipients of a transplant from a seropositive donor are especially at risk for primary infection as CMV can be transmitted by the transplant. In patients treated with antilymphocyte globulin (ALG) preparations for rejection, CMV disease is diagnosed three to four times more frequent than in patients without ALG therapy. Prevention of CMV infection is possible by selecting CMV seronegative donors for seronegative recipients. Active and passive immunization does not prevent CMV infection after renal transplantation, but immunoprophylaxis may result in less severe CMV disease. Effective treatment of clinical overt CMV disease is possible with the new guanine analogue ganciclovir. However, the use of this drug is associated with neutropenia, especially in patients with compromised kidney function. More pharmacokinetic data are needed to determine optimal dosing schemes. PMID- 2550412 TI - Cytomegalovirus infections in heart and heart-lung transplant recipients. AB - Of the first 250 heart and 41 heart-lung transplant recipients in the Papworth series, 244 who survived for longer than one month after transplantation were studied for evidence of cytomegalovirus (CMV) infection by serological methods. Overall, 43 (17.6%) patients acquired primary CMV infection and 76 (31.1%) had evidence of CMV reactivation or reinfection. The source of CMV infection was found to be important. CMV infection was more severe when acquired from the donor organ than from blood or blood products and all seven fatal primary CMV infections were associated with CMV antibody-positive donors. Thus, we now have a CMV matching policy in our unit; whenever possible organs from CMV antibody positive donors are not given to CMV antibody-negative recipients. Those CMV antibody-positive recipients who received organs from CMV antibody-positive donors had more severe CMV reactivation or reinfection than those who received organs from CMV antibody-negative donors. The severity of CMV disease was found to be dependent on the degree of immunosuppression patients received and heart lung transplant recipients had more serious CMV infections than heart transplant patients. The severity of CMV infections in heart-lung transplant recipients was modified in the later part of the series by the use of prophylactic anti-CMV hyperimmune globulin and treatment of CMV infection with ganciclovir. While 75% of heart-lung transplant recipients who did not receive this treatment died of primary donor-acquired CMV infection, all three comparable patients given CMV hyperimmune globulin and ganciclovir survived. The outcome of CMV infections in the Papworth heart and heart-lung transplant recipients is compared with that in other comparable transplant centres. PMID- 2550413 TI - Ganciclovir therapy in iatrogenically immunosuppressed patients with cytomegalovirus disease. AB - A study of ganciclovir used on compassionate grounds in the treatment of severe cytomegalovirus disease in iatrogenically immunosuppressed and AIDS patients in Europe, commenced in June 1985. The results of 120 iatrogenically immunosuppressed patients treated between May 1986 and February 1988 are reported. Patients presented with systemic infection (58), pneumonia (58), retinitis (9) and infection of other organs (9). There was a favourable clinical response (as rated by the local physician) in 40 of 57 (70%) evaluable patients with systemic infection, 38/58 (65%) with pneumonia, 6/8 (75%) with retinitis, and 4/9 (44%) of those with CMV infection of other organs. Of particular note was the relatively high rate of short term survival (up to 12 days post therapy) in bone marrow transplant patients with CMV pneumonia, 10/19 (52%), compared to the 10% survival reported by Shepp et al., (1985). Adverse events were frequent (26%) and haematological effects necessitated discontinuation of treatment in 13/120 (11%) patients. The use of ganciclovir should be confined, therefore, to the treatment of severe cytomegalovirus disease. PMID- 2550414 TI - Cytomegalovirus pneumonia: the use of ganciclovir in marrow transplant recipients. AB - Sixteen bone marrow transplant recipients with cytomegalovirus pneumonia diagnosed by rapid, non-invasive methods were treated with ganciclovir. Seven patients survived the acute infection and there were five long term survivors. Excellent in-vivo suppression of cytomegalovirus was observed. Marrow toxicity was noted in four patients but was rapidly reversible and not life threatening. Clinical features common to surviving patients included good clinical condition, insidious development of infection and evidence of normal alveolar gas exchange. The fulminant onset of symptoms, radiographic abnormalities and hypoxaemia were characteristic of non-survivors. These results offer some encouragement towards further study of ganciclovir for the early treatment of cytomegalovirus pneumonia. To identify such patients, the use of rapid diagnostic methods and aggressive viral surveillance is recommended. Convincing evidence for the efficacy of this drug will only emerge from randomized prospective studies. PMID- 2550415 TI - Protection of piperacillin and ticarcillin from beta-lactamase hydrolysis by tazobactam (YTR-830) and clavulanic acid. PMID- 2550416 TI - Fluconazole treatment of Candida glabrata peritonitis. PMID- 2550417 TI - Lack of CSF-1 receptor message in Reed-Sternberg cells. AB - The histogenesis of the Reed-Sternberg (R-S) cell in Hodgkin's disease is uncertain. Some have suggested that it is a derivative of the monocyte/macrophage lineage. To explore this possibility, we have searched for the presence of mRNA corresponding to the c-fms proto-oncogene, a marker for cells of the monocyte/macrophage lineage which encodes the colony-stimulating factor-1 receptor. In situ hybridization was performed using a single-stranded c-fms complementary RNA (cRNA) to probe R-S cells, lymphocytes, and eosinophils from touch imprints of a lymph node from a 12-year-old boy with mixed cellularity Hodgkin's disease in relapse. The probe was synthesized from a bacterial plasmid, pSM3, into which a portion of v-fms (a viral-derived oncogene) had been inserted. The plasmid was linearized with a restriction endonuclease, and 35S-labeled cRNA was synthesized from the DNA template using T3 RNA polymerase and the nucleotide analog [35S]UTP. Positive control hybridizations were obtained with the human acute promyelocytic cell line HL-60 induced to monocyte differentiation. R-S cells were clearly negative, supporting a cell of origin other than the monocyte. In situ hybridization is a potentially powerful method for exploring differentiation and assigning cell lineage in R-S cells. PMID- 2550418 TI - Complete remission of paraneoplastic sensorimotor neuropathy: a case associated with small-cell lung cancer responsive to chemotherapy, plasma exchange, and radiotherapy. AB - A 61-year-old man was found to have small-cell lung cancer following a 1-year history of a progressive peripheral sensorimotor neuropathy. The neuropathy initially improved following chemotherapy, but subsequently progressed to the point of respiratory failure. Treatment with plasma exchange, additional chemotherapy, and radiotherapy resulted in a sustained complete tumor remission and neurologic recovery. The role of plasma exchange is unclear, but its use should be considered in cases of severe sensorimotor neuropathy unresponsive to antineoplastic treatment. PMID- 2550419 TI - Protein sorting between the outer and inner mitochondrial membranes: submitochondrial localization of cytochrome c1 whose presequence is replaced by the amino-terminal region of a 70 kDa outer membrane protein. AB - The amino-terminal region of a 70 kDa mitochondrial outer membrane protein of yeast and the presequence of cytochrome c1, an inner membrane protein exposed to the intermembrane space, are thought to be responsible for localizing the proteins in their final destinations after synthesis in the cytosol. Gene fusion experiments were used to identify signals that are responsible for protein sorting between the outer and inner mitochondrial membranes. The submitochondrial localization of cytochrome c1 whose presequence was replaced by the amino terminal region of the 70 kDa mitochondrial outer membrane protein has been investigated. We have also used an in vivo complementation assay to determine whether or not a 70k-cyt c1 fusion protein is functional. Both the first half and all of the presequence of cytochrome c1 can be replaced by the amino-terminal 12 or 29 residues of the 70 kDa protein for transport to the inner membrane and functional assembly into succinate-cytochrome c reductase. However, replacements by the amino-terminal 61 residues of the 70 kDa protein result in exclusive localization of the fusion proteins to the outer membrane, and the fusions cannot be assembled into the enzyme complex. These data indicate that a mitochondrial targeting signal alone is sufficient to direct cytochrome c1 of mature size to the inner membrane. PMID- 2550420 TI - Chemical deglycosylation of hen ovomucoid: protective effect of carbohydrate moiety on tryptic hydrolysis and heat denaturation. AB - Hen ovomucoid was chemically deglycosylated by treatment with trifluoromethanesulfonic acid at 0 degrees C for 60 min. About 75 mol% of the carbohydrate moiety was removed from the glycoprotein without changing its amino acid composition, and its trypsin inhibitory activity and immunoreactivity with specific antibodies remained unchanged. The deglycosylated ovomucoid was inactivated and degraded easily by an excess amount of trypsin, whereas the native glycoprotein was not. Furthermore, the biological and immunological activities of the deglycosylated ovomucoid were lowered by heat treatment more easily than those of the native ovomucoid. These results suggest that the carbohydrate moiety of ovomucoid contributes to the stability of the ovomucoid molecule against tryptic hydrolysis and heat denaturation. PMID- 2550421 TI - Structure and expression of mouse apolipoprotein E gene. AB - The mouse apolipoprotein E gene was isolated from a genomic library by screening with a cDNA probe. DNA including apolipoprotein E gene plus segments 2.5 kilobases upstream and 0.3 kilobase downstream of the coding region was transfected into NIH3T3 cells. The cells expressed the same-size apolipoprotein E mRNA and protein as those produced by mouse endogenously. The nucleotide sequence of the gene plus 5' and 3' flanking regions (one kilobase each) was determined. The sequence of the mouse apoliprotein E gene was highly homologous to that of the rat gene, not only in the coding regions but also in the non-coding and intron regions. The mouse and the human apolipoprotein E genes were homologous in the 5' proximal flanking region up to about 200 nucleotides as well as in the four exons. This proximal region was highly conserved for the genes of mouse, rat and human; the relative positions of the "TATA box" and the two copies of "GC box" were identical. PMID- 2550422 TI - Regulation of rhodopsin dephosphorylation by arrestin. AB - We have characterized the opsin phosphatase activities in extracts of rod outer segments and determined their relationship to known protein phosphatases. The opsin phosphatase activity in the extracts was not due to protein phosphatases 1, 2B, or 2C because it was neither stimulated by Mg2+ or Ca2+/calmodulin nor inhibited by protein phosphatase inhibitors-1 or -2. Opsin phosphatase activity in rod outer segment extracts was potently inhibited by okadaic acid (IC50 approximately 10 nM), a preferential inhibitor of protein phosphatase 2A. Moreover, during chromatography on DEAE-Sepharose, the opsin phosphatase activity co-eluted with three peaks of protein phosphatase 2A activity, termed protein phosphatases 2A0, 2A1, and 2A2. The opsin phosphatase activity of each peak was stimulated by polylysine, a known activator of protein phosphatase 2A. Finally, treatment of rod outer segment extracts with 80% ethanol at room temperature converted the activity from a high molecular weight form characteristic of the protein phosphatase 2A0, 2A1, and 2A2 species to a low molecular weight form characteristic of the protein phosphatase 2A catalytic subunit. We conclude that protein phosphatase 2A is likely to be the physiologically relevant rhodopsin phosphatase. The 48-kDa rod outer segment protein arrestin (S-antigen) was found to inhibit the dephosphorylation of freshly photolyzed rhodopsin by protein phosphatase 2A but did not inhibit the dephosphorylation of unbleached rhodopsin. Arrestin has no effect on the dephosphorylation of phorphorylase a, indicating that the effect was substrate-directed. It appears that dephosphorylation of the photoreceptor protein phosphorhodopsin occurs only after decay of the photoactivated protein and that this may be regulated in vivo by arrestin. The binding of arrestin to photolyzed phosphorylated rhodopsin, i.e. the binding of a regulatory protein to a protein phosphatase substrate to form a complex resistant to dephosphorylation represents a novel mechanism for the regulation of protein phosphatase 2A. PMID- 2550423 TI - Characterization of the flavoprotein moieties of NADPH-sulfite reductase from Salmonella typhimurium and Escherichia coli. Physicochemical and catalytic properties, amino acid sequence deduced from DNA sequence of cysJ, and comparison with NADPH-cytochrome P-450 reductase. AB - NADPH-sulfite reductase flavoprotein (SiR-FP) was purified from a Salmonella typhimurium cysG strain that does not synthesize the hemoprotein component of the sulfite reductase holoenzyme. cysJ, which codes for SiR-FP, was cloned from S. typhimurium LT7 and Escherichia coli B, and both genes were sequenced. Physicochemical analyses and deduced amino acid sequences indicate that SiR-FP is an octamer of identical 66-kDa peptides and contains 4 FAD and 4 FMN per octamer. Potentiometric titrations of SiR holoenzyme, SiR-FP, and FMN-depleted SiR-FP yielded the following redox potentials for the prosthetic groups at pH 7.7: E'1 (FMNH./FMN) = -152 mV; E'2 (FMNH2/FMNH.) = -327 mV; E'3 (FADH./FAD) = -382 mV; E'4 (FADH2/FADH.) = -322 mV. Microcoulometric titration of SiR-FP at 25 degrees C yielded data which were in full agreement with these potentials. Spectroscopic and catalytic studies of native SiR-FP and of SiR-FP depleted of FMN support the following electron flow sequence: NADPH----FAD----FMN. FMN can then contribute electrons to the hemoprotein component of sulfite reductase, as well as to cytochrome c and various diaphorase acceptors. The FMN is postulated to cycle between the FMNH2 and FMNH. oxidation states during catalysis; in this sense SiR FP shares a catalytic mechanism with NADPH-cytochrome P-450 oxidoreductase. SiR FP domains involved in binding FMN, FAD, and NADPH are proposed from amino acid sequence homologies with Desulfovibrio vulgaris flavodoxin (Dubourdieu, M., and Fox, J.L. (1977) J. Biol. Chem. 252, 1453-1463) and spinach ferredoxin-NADP+ oxidoreductase (Karplus, P.A., Walsh, K.A., and Herriott, J. R. (1984) Biochemistry 23, 6576-6583). Comparison of the deduced amino acid sequences of SiR-FP and NADPH-cytochrome P-450 oxidoreductase (Porter, T. D., and Kasper, C.B. (1985) Proc. Natl. Acad. Sci. U. S.A. 82, 973-977) also showed identities that suggest these two proteins are descended from a common precursor, which contained binding regions for both FMN and FAD. PMID- 2550424 TI - Characterization of the binding of human tissue-type plasminogen activator to platelets. AB - Cell surface binding sites for the constituent proteins of the fibrinolytic system may play a role in the localization and regulation of fibrinolysis. In the present study, specific binding of recombinant human tissue-type plasminogen activator (rt-PA) to human blood platelets was identified and characterized. 125I labeled rt-PA was found to bind specifically, saturably, and reversibly to the surface of gel-filtered platelets, reaching equilibrium within 5 min at 22 degrees C. Scatchard analysis revealed a single class of binding sites. Unstimulated platelets bound 120,000 +/- 24,000 (mean +/- S.D.) molecules/platelet with an apparent Kd of 340 +/- 25 nM, whereas thrombin stimulated platelets bound 290,000 +/- 32,000 molecules/platelet with an apparent Kd of 800 +/- 60 nM. Binding of 0.1 microM 125I-rt-PA was greater than 90% reversible by a 50-fold excess of unlabeled rt-PA. Binding was not inhibited by fibrinogen or single chain urokinase-type plasminogen activator, but plasminogen partially competed for binding of 125I-rt-PA to platelets (up to 40% displacement). These findings indicate that the platelet surface possesses a large number of specific, low affinity binding sites for t-PA and provide further evidence for the role of platelets in localization and regulation of fibrinolysis. PMID- 2550425 TI - The redox state of cysteines 201 and 317 of the erythrocyte anion exchanger is critical for ankyrin binding. AB - Previous studies have demonstrated that modification of erythrocyte membrane cysteine residues via disulfide cross-briding or direct derivatization with thiol reagents promotes massive morphological, rheological, and structural changes in the cell. To determine whether disruption of the band 3-ankyrin interaction, the major membrane-cytoskeletal linkage, might contribute to the above lesions, we quantitatively measured the band 3-ankyrin interaction following modification of Cys-201 and/or Cys-317 of the cytoplasmic domain of band 3. It was observed that irreversible alkylating agents (e.g. N-ethylmaleimide or iodoacetamide and its derivatives), reversible derivatizing compounds (.e.g. p chloromercuribenzenesulfonate or glutathione), and native disulfide bond formation all blocked the ankyrin interaction. Comparison of the extent of sulfhydryl modification with the degree of inhibition of ankyrin binding further confirmed that cysteine modification was directly responsible for the inhibition. However, analysis of the site of sulfhydryl derivatization revealed that inhibition of ankyrin binding could be initiated in some cases with derivatization of Cys-201, while in other cases obstruction of Cys-317 appeared to be essential. This apparent discrepancy was resolved by demonstrating that Cys 201 of one strand of the cytoplasmic domain of band 3 dimer could disulfide bond with Cys-317 of the opposite strand, thus demonstrating that all four cysteines of the band 3 dimer are clustered at the interface between subunits. We argue that derivatization or disulfide cross-linking of these cysteines can block ankyrin binding by both conformational and steric mechanisms. PMID- 2550426 TI - Mutation of the high cysteine region of the human insulin receptor alpha-subunit increases insulin receptor binding affinity and transmembrane signaling. AB - Our previous studies indicated that amino acid residues 240-250 in the cysteine rich region of the human insulin receptor alpha-subunit constitute a site in which insulin binds (Yip, C. C., Hsu, H., Patel, R. G., Hawley, D. M., Maddux, B. A., and Goldfine, I. D. (1988) Biochem. Biophys. Res. Commun. 157, 321-329). We have now constructed a human insulin receptor mutant in which 3 residues in this sequence were altered (Thr-Cys-Pro-Pro-Pro-Tyr-Tyr-His-Phe-Gln-Asp to Thr-Cys-Pro Arg-Arg-Tyr-Tyr-Asp-Phe-Gln-Asp) and have expressed this mutant in rat hepatoma (HTC) cells. When compared with cells transfected with normal insulin receptors, cells transfected with mutant receptors had an increase in insulin-binding affinity and a decrease in the dissociation of bound 125I-insulin. Studies using solubilized receptors also demonstrated that mutant receptors had a higher binding affinity than normal receptors. In contrast, cells transfected with either mutant or normal receptors bound monoclonal antibodies against the receptor alpha-subunit with equal affinity. When receptor tyrosine kinase activity and alpha-aminoisobutyric acid uptake were measured, cells transfected with mutant insulin receptors were more sensitive to insulin than cells transfected with normal receptors. These findings lend further support therefore to the hypothesis that amino acid sequence 240-250 of the human insulin receptor alpha-subunit constitutes one site that interacts with insulin, and they indicate that mutations in this site can influence insulin receptor binding and transmembrane signaling. PMID- 2550427 TI - Self-induction of 1,25-dihydroxyvitamin D3 metabolism limits receptor occupancy and target tissue responsiveness. AB - Whole cell 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) receptor (VDR) binding assays, which measure VDR in the presence of the metabolic machinery of the cell, were used in conjunction with a cytosol binding assay for VDR to determine if self induced metabolism of 1,25-(OH)2D3 limits VDR occupancy, total VDR levels, and target cell responsiveness. Treatment of cells with 0.5 nM 1,25-(OH)2[3H]D3 for 16 h results in up-regulation of total cell VDR from 82 to 170 fmol/mg protein as measured in a cytosol binding assay. Conversely, whole cell binding assays of VDR showed a 1,25-(OH)2D3-mediated apparent down-regulation of VDR from 90 to 40 fmol/mg protein. Scatchard analysis using the cytosol binding assay demonstrated that 1,25-(OH)2D3 treatment increased total cell VDR from 93 to 154 fmol/mg protein. In contrast, Scatchard analysis with the whole cell binding assay demonstrated that 1,25-(OH)2D3 treatment resulted in reduction in total cell VDR from 100 to 64 fmol/mg protein. Initial Kd estimates with the whole cell binding assay suggested that 1,25-(OH)2D3 treatment resulted in a reduction in VDR Kd from 0.6 to 6.2 nM. This apparent reduction in the affinity of VDR for 1,25 (OH)2D3 was due to degradation of free 1,25-(OH)2[3H]D3 which occurred during whole cell saturation assay. Competitive inhibitors of 1,25-(OH)2D3 metabolism were found to reverse the apparent receptor down-regulation observed in whole cell binding assays of treated cells. In addition, the presence of competitive inhibitors amplified responses of cells to 1,25-(OH)2[3H]D3 treatment as measured by an increased occupancy of VDR by 1,25-(OH)2[3H]D3 and increased up-regulation of VDR over that observed without metabolism inhibitors. These data demonstrate that self-induced target tissue deactivation of 1,25-(OH)2D3 regulates 1,25 (OH)2D3 occupancy of VDR and ultimately the biopotency of 1,25-(OH)2D3 in target cells. PMID- 2550428 TI - An alternate promoter in the glucokinase gene is active in the pancreatic beta cell. AB - An alternate promoter in the glucokinase gene is active in the beta cell and produces a glucokinase mRNA which is longer and that has a different leader sequence and translation start site than the hepatic glucokinase mRNA. The glucokinase beta cell promoter is located at least 12 kilobases upstream from the glucokinase hepatic promoter. Transcription from the glucokinase beta cell promoter initiates over a region of 62 bases. The absence of a TATA box homology in the proximal promoter region may account for the diffuse transcriptional initiation. Translation of the beta cell glucokinase mRNA predicts a glucokinase isozyme that is different from the hepatic isozyme by 15 amino acids at the N terminus. The use of alternative promoters apparently enables the glucokinase gene to be regulated by insulin in the liver and by glucose in the beta cell, thus possibly constituting an important feedback control loop for maintaining glucose homeostasis. Alternate RNA splicing of the beta cell glucokinase mRNA predicts at least two beta cell glucokinase isoforms. An alternate splice acceptor site in the 4th exon of the glucokinase gene was identified in two glucokinase cDNAs from rat insulinoma tissue. Use of the alternate splice acceptor site results in a 51-nucleotide in frame deletion in the beta cell glucokinase mRNA and removal of 17 amino acids from a region of the protein situated between the putative glucose and ATP binding domains. Analysis of the pattern of RNA splicing in tissues containing beta cells indicates that the splice acceptor site utilized in producing hepatic glucokinase mRNA is also utilized in the beta cell. PMID- 2550429 TI - Degradation of ornithine decarboxylase in reticulocyte lysate is ATP-dependent but ubiquitin-independent. AB - Reticulocyte lysate contains all the components of the ubiquitin-dependent proteolytic system. Several proteins are degraded in reticulocyte lysate in a ubiquitin-dependent manner. However, none of the proteins studied has a short intracellular half-life. We have investigated the degradation of ornithine decarboxylase (ODC), one of the most labile proteins in mammalian cells. ODC is efficiently degraded in reticulocyte lysate depleted of the ubiquitin activating enzyme, E1, in fraction II of reticulocyte lysate completely lacking ubiquitin, and in fraction II depleted of the entire complex of enzymes responsible for the ligation of ubiquitin to target proteins. The degradation of ODC is ATP dependent. Therefore, our results demonstrate that in addition to the ubiquitin dependent proteolytic pathway, reticulocyte lysate contains at least one additional ATP-dependent proteolytic pathway. In vitro synthesized ODC served as a substrate in the present degradation study. Its successful utilization establishes a general strategy for investigating the degradation of short-lived proteins (for which a corresponding cDNA is available), that constitute a very small fraction of cellular proteins and for which purification is difficult or impossible. In contrast to ODC synthesized in vitro, that isolated from cells was not degraded by the reticulocyte lysate degradation system, suggesting that post translational modifications may be involved in regulating ODC degradation. PMID- 2550430 TI - Erythropoietin. Receptor characteristics during the ontogeny of hamster yolk sac erythroid cells. AB - Erythropoietin (Epo) binds specifically to receptors on the surface membrane of responsive erythroid cells. In search of ontogenic changes in Epo receptor behavior, we studied characteristics of specific binding to hamster yolk sac erythroid cells during hamster ontogeny. We detected receptors specific for Epo on these cells throughout the duration of their intravascular existence (hamster gestational days 8 through 13). These receptors are saturable at an Epo concentration of 1.2 nM in the incubation medium. Attainment of equilibrium of binding prior to hormone internalization, a requirement for receptor binding assays, was possible at 10 degrees C but not at 37 degrees C. Hence, all incubations of cells with Epo were carried out at 10 degrees C. Data on specific binding analyzed by the method of Scatchard demonstrated that yolk sac erythroid cells possess a single class of Epo receptors at each stage of gestation examined. Binding affinity and numbers of receptors per cell change as ontogeny progresses: Kd (the dissociation constant) increases, a phenomenon observed in other differentiating cell systems, whereas the number of receptors per cell peaks on gestational day 10. The variability in number of receptors per cell is consonant with up and down regulation controlled by Epo availability. We propose that the progressive increase in Kd might be best explained by ontogenic changes in cell membrane structure contiguous to the receptors themselves. PMID- 2550431 TI - Evidence from extended X-ray absorption fine structure and site-specific mutagenesis for zinc fingers in UvrA protein of Escherichia coli. AB - The UvrA protein is the damage recognition subunit of the Escherichia coli repair enzyme ABC excision nuclease. Sequence analysis of this 940-amino acid protein revealed two regions of sequence homology to the zinc finger motif found in many DNA binding proteins. Physical and chemical analyses indicate about 2 zinc atoms/molecule. We have used extended x-ray absorption fine structure analysis to demonstrate that each of these zinc atoms is coordinated with 4 cysteine residues at a distance of 2.32 +/- 0.2 A. Substitution of one of the cysteines by a histidine, a serine, or an alanine in one of the potential finger sites resulted in a respective decrease in complementing activity. We thus conclude that the two zinc fingers identified by sequence analysis do indeed have zinc finger structure in UvrA protein. PMID- 2550432 TI - IIb group metal ions (Zn2+, Cd2+, Hg2+) stimulate glucose transport activity by post-insulin receptor kinase mechanism in rat adipocytes. AB - Zn2+ (1 mM), Cd2+ (1 mM), and Hg2+ (0.1 mM) belonging to the IIb group in the periodic table stimulated glucose transport activity and cAMP phosphodiesterase in rat adipocytes. The stimulation of glucose transport was due to the translocation of glucose transporters from the intracellular site to the plasma membrane. However, in intact adipocytes none of these ions stimulated insulin receptor kinase activity or phosphorylation of the 95-kDa subunit of insulin receptor or 170- or 60-kDa proteins at the tyrosyl residues. These proteins were markedly phosphorylated by addition of 0.3 nM insulin which stimulated glucose transport activity as effectively as these metal ions. These results indicate that Zn2+, Cd2+, and Hg2+ mimic insulin action by a post-receptor/kinase mechanism. PMID- 2550433 TI - Human myelogenous leukemia cell line HL-60 cells resistant to differentiation induction by retinoic acid. Decreased content of glycosphingolipids and granulocytic differentiation by neolacto series gangliosides. AB - We have recently reported that neolacto series gangliosides (NeuAc-nLc) are increased during granulocytic differentiation of human myelogenous leukemia cell line HL-60 cells induced by retinoic acid and that HL-60 cells are differentiated into mature granulocytes when the cells are cultivated with NeuAc-nLc (Nojiri, H., Kitagawa, S., Nakamura, M., Kirito, K., Enomoto, Y., and Saito, M. (1988) J. Biol. Chem. 263, 7443-7446). In contrast to these wild-type-HL-60 cells, HL-60 cells resistant to differentiation induction by retinoic acid showed a markedly decreased content of gangliosides, especially NeuAc-nLc, and did not show any increase in the content of gangliosides when cultivated with retinoic acid. Neutral glycosphingolipids, the precursors of gangliosides, were not accumulated in these resistant cells. When retinoic acid-resistant HL-60 cells were cultivated in the presence of NeuAc-nLc, the cells were found to be differentiated into mature granulocytes on morphological and functional criteria. The differentiation of cells was dependent on the concentration of gangliosides and was accompanied by inhibition of cell growth. Wild-type HL-60 cells differentiated by NeuAc-nLc showed the changes in ganglioside composition, which were similar to those in wild-type HL-60 cells differentiated by retinoic acid; among the gangliosides changed, 2----3 sialylparagloboside and 2----3 sialylnorhexaosylceramide were increased. These findings suggest (a) that the synthesis of particular NeuAc-nLe molecules is an important step for retinoic acid-induced granulocytic differentiation and this step could be bypassed or replaced by exogenous NeuAc-nLc, and (b) that the defective synthesis of particular NeuAc-nLc molecules is responsible for the failure of differentiation induction in retinoic acid-resistant HL-60 cells by retinoic acid. PMID- 2550434 TI - A study of drug-induced topoisomerase II-mediated DNA lesions on episomal chromatin. AB - A well defined extrachromosomal DNA element, referred to as an episome (Ostrowski, M., Richard-Foy, H., Wolford, R., Berard, D., and Hager, G. (1983) Mol. Cell. Biol. 3, 2045-2057), was employed as a target for the topoisomerase II inhibitors amsacrine and teniposide. Both drugs have distinct mechanisms of action in cleaving the episome, as defined by topological forms conversion assays. The concentration ranges required to measure episomal cleavage are similar. The onset of damage induced by amsacrine begins within 1 min and is maintained at that level for at least 1 h. Teniposide induces damage that peaks between 30 and 60 min. The amsacrine-induced damage is only partially reversible, whereas teniposide-induced damage is almost completely reversible. Sites of specific cleavage are quite dissimilar. Multiple cleavage sites are formed in the episomal regulatory regions after amsacrine treatment, whereas a single cleavage in the regulatory region and one outside this region are found after teniposide treatment. Transcriptional activation using dexamethasone does not change the amount or site preference of episomal cleavage induced by either agent. Damage to the episome was quantitatively compared with damage produced in genomic DNA between 500 and 24,000 rad equivalents. The study showed that amsacrine has a significant (33-38-fold) preference for episomal DNA over genomic DNA. PMID- 2550435 TI - The activation of bovine protein C by factor Xa. AB - A complex composed of factor Xa and phospholipid vesicles assembled in the presence of calcium ions catalyzes a discrete cleavage of the heavy chain of bovine protein C that is indistinguishable from that produced by thrombin as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This cleavage generates an active site capable of hydrolyzing small substrates and inactivating factor Va function in the prothrombinase complex. Activation of protein C by factor Xa requires both calcium ions and phospholipid vesicles and proceeds at a rate an order of magnitude greater than that observed for alpha thrombin in solution. gamma-Carboxyglutamic acid-domainless protein C is not activated by factor Xa, consistent with the requirement for phospholipid and distinguishing this reaction from protein C activation by thrombin. Thrombomodulin serves as a cofactor for the factor Xa-catalyzed reaction, forming a 1:1 complex with factor Xa (apparent Kd = 5.7 X 10(-10) M) and stimulating the saturated rate of protein C activation by factor Xa (kcat = 149 min-1) to levels comparable with the thrombin-thrombomodulin complex. Protein C activation by factor Xa is not inhibited by the specific thrombin inhibitor dansyl-N-(3-ethyl 1,5-pentanediyl)amide but is inhibited by antithrombin III, tripeptide chloromethyl ketones, and the monoclonal antibody alpha-BFX-2b that is highly specific for factor Xa. These data indicate that thrombomodulin is promiscuous in its role as a cofactor and suggest the existence of an alternative pathway for protein C activation in vivo. PMID- 2550436 TI - Identification of a Ni- and Fe-containing cluster in Rhodospirillum rubrum carbon monoxide dehydrogenase. AB - Methyl viologen-oxidized carbon monoxide dehydrogenase (CODH) from Rhodospirillum rubrum exhibits complex EPR. Comparison to EPR of oxidized apo-CODH (CODH from which Ni is lacking) leads to the identification of signals whose intensity is correlated with the presence of Ni. 61Ni labeling observably broadens the sharpest feature of these signals, as does 57Fe. R. rubrum CODH thus contains a cluster containing both Ni and Fe. The EPR associated with this cluster is unlike any EPR previously attributed to Ni-containing prosthetic groups in other CODH enzymes or Ni-containing hydrogenases. The CO-analogue, CN-, perturbs the EPR signals that are attributed to the Ni-Fe species. PMID- 2550438 TI - Oscillating actin polymerization/depolymerization responses in human polymorphonuclear leukocytes. AB - Leukotriene B4 and platelet-activating factor induced a rapidly oscillating actin polymerization/depolymerization response in human polymorphonuclear leukocytes. N Formylpeptides were deficient in the ability to induce these oscillations. Flow cytometric analysis of filamentous actin verified that all cells were synchronously responding in this cyclic manner. The hypothesis was tested that these oscillations were analogous to chemical oscillations, i.e. oscillations of intermediate species in chemical systems that are far from equilibrium (Epstein, I. R., Kustin, K., DeKepper, P., and Orban, M. (1983) Sci. Am. 248, 112). Actin polymerization/depolymerization cycles were terminated by adding receptor antagonist a few seconds after initiation of the response by agonists. Thus the oscillations did not represent chemical oscillations that hypothetically could result from a rapid jump of the intracellular milieu to a state far from equilibrium. Rather, continued occupancy of receptors and/or occupancy of new receptors was required to sustain the oscillations. This suggested that the oscillations resulted from regulated polymerization and depolymerization pathways. In simultaneous measurements of actin-associated right angle light scatter and intracellular calcium, no calcium oscillations were detected. Thus, cycles of actin polymerization/depolymerization were not regulated by calcium oscillations. PMID- 2550437 TI - Tumor necrosis factor acts synergistically with autocrine interferon-beta and increases interferon-beta mRNA levels in human fibroblasts. AB - Medium of untreated human FS-4 foreskin fibroblasts contained a factor which, upon the addition of exogenous tumor necrosis factor (TNF), inhibited encephalomyocarditis virus replication when neither medium alone nor TNF alone were effective. This antiviral activity was abolished by a monoclonal antibody to human interferon (IFN)-beta, suggesting that the active component in the medium from untreated FS-4 cells was IFN-beta, present at subeffective concentrations. In addition, we show that untreated FS-4 cells contain IFN-beta mRNA, demonstrable by the highly sensitive polymerase chain reaction after reverse transcription. Treatment of FS-4 cells with TNF produced an approximately 16-fold increase in the steady-state level of IFN-beta mRNA. Our results support the conclusion that autocrine IFN-beta is secreted by untreated normal fibroblasts and that TNF can enhance the production of autocrine IFN-beta by increasing the level of IFN-beta mRNA. Our study also demonstrates that subeffective concentrations of autocrine IFN-beta, which escape detection in conventional assays, are sufficient to produce a strong synergistic action with TNF. PMID- 2550439 TI - Biotinylated parathyroid hormone as a probe for the parathyroid hormone receptor. Structure-function analysis and detection of specific binding to cultured bone cells by flow cytometry. AB - The synthetic bovine parathyroid hormone (PTH) analog (Nle8, Nle18, Tyr34) bovine PTH(1-34)amide (bPTH(1-34)amide) was reacted with biotinyl-epsilon aminocaproic acid-N-hydroxysuccinimide under conditions which yielded five isoforms which were fractionated by a combination of reversed phase and ion-exchange chromatography. These reaction products were analyzed by automated Edman degradation in a manner which allowed us to specify the location and number of biotin residues on picomole quantities of hormone. The ability of each of these isoforms to induce a rise in intracellular cAMP in the ROS 17/2.8 cell line allowed us to evaluate the effect on function of biotinylation at different residues. Derivatized PTH molecules which contained a single biotin at either lysine 13, lysine 26, or lysine 27 possessed full biological activity. However, bioactivity was significantly reduced when position 13 plus either lysine 26 or 27 were biotinylated. Biological activity was lost when all 3 lysine residues were biotinylated. Biotinylation of the alpha-NH2 group of alanine at the NH2 terminus also resulted in a total loss of activity. Hence, unlike the effect of altering the alanine at position 1, modification of a single lysine residue at positions 13, 26, and 27 has a less critical effect on biological activity of the molecule. However, biotinylation of all three lysines results in a biologically inert PTH derivative and suggests that changes in isoelectric point, hydrophobicity, or tertiary structure may strongly influence hormone function. A fully bioactive mixture of isoforms was used to detect receptors on ROS 17/2.8 cells by flow cytometry using fluorescein isothiocyanate-avidin as a fluorescent indicator. Binding to cell surface receptors was saturable and could be inhibited by native bPTH(1-34) but not by transforming growth factor beta, calcitonin or insulin. Moreover, PTH receptors could also be detected on primary cultures of human bone cells and human fibroblasts. PMID- 2550441 TI - Chicken and Xenopus mannose 6-phosphate receptors fail to bind insulin-like growth factor II. AB - The recent demonstration that a single mammalian receptor protein binds both mannose 6-phosphate (Man-6-P) and insulin-like growth factor II (IGF-II) with high affinity has suggested a multifunctional physiological role for this receptor, possibly including signal transduction. In order to better understand the functions of this receptor, we have investigated the properties of Man-6-P receptors from non-mammalian species. Receptors were affinity-purified from Triton X-100 extracts of total membranes from Xenopus and chicken liver as well as rat placenta using pentamannosyl 6-phosphate-Sepharose. The Man-6-P receptor was adsorbed to the pentamannosyl 6-phosphate-Sepharose and specifically eluted by Man-6-P in all three species, as evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by silver staining. When the purified receptors from these three species were cross-linked to 125I-IGF-II with disuccinimidyl suberate, only receptors isolated from rat membranes were affinity labeled. To further evaluate the properties of these Man-6-P receptors, binding of 125I-rat-IGF-II and 125I-chicken Tyr-Gly-Thr-Ala-IGF-II to purified receptors from Xenopus, chicken, and rat was evaluated by polyethylene glycol precipitation. Only the rat Man-6-P receptor exhibited detectable binding of 125I IGF-II. These data suggest that the emergence of a high affinity IGF-II binding site on the Man-6-P receptor occurred in evolution after the divergence of mammals from other vertebrates. Thus, the biological actions of IGF-II in chickens and frogs appear to be initiated by the type I IGF receptor. PMID- 2550440 TI - Identification of the ral and rac1 gene products, low molecular mass GTP-binding proteins from human platelets. AB - Identification of the GTP-binding proteins from human platelet particulate fractions was attained by their purification via successive column chromatography steps followed by amino acid sequencing. To enhance the likelihood of identifying the GTP-binding proteins, two assays were employed to monitor GTP-binding activities: (i) guanosine 5'-(3-O-[35S]thio)triphosphate (GTP gamma S)-binding followed by rapid filtration and ii) [alpha-32P]GTP-binding following sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electroblotting onto nitrocellulose membranes. The latter assay permitted the isolation of a 28-kDa GTP-binding protein that bound [alpha-32P]GTP prominently but was only poorly detected with the GTP gamma S-binding assay. The amino acid sequences of three peptide fragments derived from the 28-kDa protein were identical to regions of the amino acid sequence deduced from a simian ral cDNA with the exception of one conservative substitution (Asp147----Glu). A full length human ral cDNA was isolated from a placental cDNA library, and its deduced amino acid sequence, compared with simian ral, also contained the Asp----Glu substitution along with two other substitutions and an additional three NH2-terminal amino acids. In addition to the 28-kDa protein, two distinct 25-kDa GTP-binding proteins were purified from platelets. One of these proteins has been previously characterized as G25K, an abundant low molecular mass GTP-binding protein. Partial amino acid sequence obtained from the second unidentified 25-kDa protein indicates that it is the product of the rac1 gene; a member of a newly identified gene family which encode for low molecular mass GTP-binding proteins (Didsbury, J., Weber, R.F., Bokoch, G. M., Evans, T., and Snyderman, R. (1989) J. Biol. Chem. 264, 16378 16382). These results identify two new GTP-binding proteins in human platelets, ral, the major protein that binds [alpha-32P]GTP on nitrocellulose transfers, and rac1, a substrate for botulinum C3 ADP-ribosyltransferase. PMID- 2550442 TI - Characterization of an amsacrine-resistant line of human leukemia cells. Evidence for a drug-resistant form of topoisomerase II. AB - HL-60/AMSA is a human leukemia cell line that is 100 times more resistant to the cytotoxic actions of the antineoplastic, topoisomerase II-reactive DNA intercalating acridine derivative amsacrine (m-AMSA) than is its parent HL-60 line. HL-60/AMSA cells are minimally resistant to etoposide, a topoisomerase II reactive drug that does not intercalate. Previously we showed that HL-60 topoisomerase II activity in cells, nuclei, or nuclear extracts was sensitive to m-AMSA and etoposide, while HL-60/AMSA topoisomerase II was resistant to m-AMSA but sensitive to etoposide. Now we show that purified topoisomerase II from the two cell lines exhibits the same drug sensitivity or resistance as that in the nuclear extracts although the magnitude of the m-AMSA resistance of HL-60/AMSA topoisomerase II in vitro is not as great as the resistance of the intact HL 60/AMSA cells. In addition HL-60/AMSA cells are cross-resistant to topoisomerase II-reactive intercalators from the anthracycline and ellipticine families and the pattern of sensitivity or resistance to the cytotoxic actions of the various topoisomerase II-reactive drugs is paralleled by topoisomerase II-reactive drug induced DNA cleavage and protein cross-link production in cells and the production of drug-induced, topoisomerase II-mediated DNA cleavage and protein cross-linking in isolated biochemical systems. In addition to its lowered sensitivity to intercalators, HL-60/AMSA differed from HL-60 in 1) the susceptibility of its topoisomerase II to stimulation of DNA topoisomerase II complex formation by ATP, 2) the catalytic activity of its topoisomerase II in an ionic environment chosen to reproduce the environment found within the living cell, and 3) the observed restriction enzyme pattern on a Southern blot probed with a cDNA for human topoisomerase II. These data indicate that an m-AMSA resistant form of topoisomerase II contributes to the resistance of HL-60/AMSA to m-AMSA and to other topoisomerase II-reactive DNA intercalating agents. The drug resistance is associated with additional biochemical and molecular alterations that may be important determinants of cellular sensitivity or resistance to topoisomerase II-reactive drugs. PMID- 2550443 TI - Redox intermediates of Desulfovibrio gigas [NiFe] hydrogenase generated under hydrogen. Mossbauer and EPR characterization of the metal centers. AB - The hydrogenase (EC 1.2.2.1) of Desulfovibrio gigas is a complex enzyme containing one nickel center, one [3Fe-4S] and two [4Fe-4S] clusters. Redox intermediates of this enzyme were generated under hydrogen (the natural substrate) using a redox-titration technique and were studied by EPR and Mossbauer spectroscopy. In the oxidized states, the two [4Fe-4S]2+ clusters exhibit a broad quadrupole doublet with parameters (apparent delta EQ = 1.10 mm/s and delta = 0.35 mm/s) typical for this type of cluster. Upon reduction, the two [4Fe-4S]1+ clusters are spectroscopically distinguishable, allowing the determination of their midpoint redox potentials. The cluster with higher midpoint potential (-290 +/- 20 mV) was labeled Fe-S center I and the other with lower potential (-340 +/- 20 mV), Fe-S center II. Both reduced clusters show atypical magnetic hyperfine coupling constants, suggesting structural differences from the clusters of bacterial ferredoxins. Also, an unusually broad EPR signal, labeled Fe-S signal B', extending from approximately 150 to approximately 450 mT was observed concomitantly with the reduction of the [4Fe-4S] clusters. The following two EPR signals observed at the weak-field region were tentatively attributed to the reduced [3Fe-4S] cluster: (i) a signal with crossover point at g approximately 12, labeled the g = 12 signal, and (ii) a broad signal at the very weak-field region (approximately 3 mT), labeled the Fe-S signal B. The midpoint redox potential associated with the appearance of the g = 12 signal was determined to be -70 +/- 10 mV. At potentials below -250 mV, the g = 12 signal began to decrease in intensity, and simultaneously, the Fe-S signal B appeared. The transformation of the g = 12 signal into the Fe-S signal B was found to parallel the reduction of the two [4Fe-4S] clusters indicating that the [3Fe-4S]o cluster is sensitive to the redox state of the [4Fe-4S] clusters. Detailed redox profiles for the previously reported Ni-signal C and the g = 2.21 signal were obtained in this study, and evidence was found to indicate that these two signals represent two different oxidation states of the enzyme. Finally, the mechanistic implications of our results are discussed. PMID- 2550444 TI - The phage T4 uvs Y recombination protein stabilizes presynaptic filaments. AB - The bacteriophage T4 uvsY protein is required for efficient recombination in T4 infected Escherichia coli cells. Previous in vitro work has shown that the purified uvsY protein is an accessory protein; it stimulates homologous pairing catalyzed by the phage uvsX protein (a RecA-like recombinase) under certain conditions. We show here that this effect can be traced, at least in part, to a UvsY-dependent stabilization of uvsX protein-single-stranded DNA complexes. These presynaptic filaments are one of the early obligatory intermediates in the strand exchange reaction between homologous single- and double-stranded DNAs. The mechanism of filament stabilization seems to involve a slower loss of UvsX subunits. A model that accounts for the data is presented in which both recombination proteins are incorporated into the presynaptic filament. PMID- 2550445 TI - Cyclic GMP-dependent stimulation of the membrane-bound insulin-sensitive cAMP phosphodiesterase from rat adipocytes. AB - The insulin-sensitive cAMP phosphodiesterase (phosphodiesterase) in rat adipocytes is a membrane-bound low Km enzyme that can be recovered in a crude microsomal fraction (Fraction P-2). The action of this enzyme to hydrolyze cAMP is known to be inhibited by cGMP; nevertheless, it was found in our present study that under selected conditions, the enzyme can also be stimulated by cGMP as well as some other nucleotide derivatives. The maximum cGMP-dependent stimulation was observed when the enzyme in Fraction P-2 was incubated with 10 microM cGMP for 5 20 min at 37 degrees C in the presence of Mg2+, washed, and then assayed in the absence of added cGMP. The level of this stimulation was close to, but less than, that achieved by insulin in intact cells. The actions of the cGMP- and insulin stimulated enzymes to hydrolyze labeled cAMP were inhibited in an identical manner by cilostamide (Ki = 0.10 microM), griseolic acid (Ki = 0.19 microM), unlabeled cAMP (Km = 0.20 microM), and cGMP (Ki = 0.16 microM), all added to the assay system. Also, the basal, insulin-stimulated, and cGMP-activated enzymes were identically inhibited by a polyclonal antibody raised against a purified membrane-bound low Km phosphodiesterase from bovine adipose tissue. When the same antibody was used for the Western blot analysis of Fraction P-2, it immunoreacted with a single band of protein (165 kDa). These observations indicate that the insulin-sensitive phosphodiesterase in rat adipocytes can be stimulated with 10 microM cGMP and that this stimulation is detectable only after the nucleotide has been eliminated since the enzyme would be strongly inhibited by the nucleotide if the latter exists in the assay system. It is proposed that the insulin-sensitive phosphodiesterase, which is often referred to as a Type IV enzyme, is functionally similar to the Type II enzymes that are known to be stimulated by a low concentration of cGMP and inhibited by higher concentrations of the same nucleotide. PMID- 2550446 TI - Protein-DNA interactions at recognition sites for the dioxin-Ah receptor complex. AB - Gel retardation analyses reveal a cluster of six binding sites for the liganded Ah receptor within a 700-base pair DNA domain upstream of the mouse CYP1A1 gene. The nucleotide sequences of the binding sites define a consensus recognition motif for the liganded receptor. The consensus motif is not symmetric. Alteration of the consensus motif produces a decrease in the receptor-DNA interaction. The ligand receptor binds as a monomer to its recognition motif and preferentially binds to double-stranded DNA. These observations reveal apparent differences between 2,3,7,8-tetrachlorodibenzo-p-dioxin and steroid hormones in their respective mechanisms of action. PMID- 2550447 TI - Regulation of calcineurin by phosphorylation. Identification of the regulatory site phosphorylated by Ca2+/calmodulin-dependent protein kinase II and protein kinase C. AB - The site in calcineurin, the Ca2+/calmodulin (CaM)-dependent protein phosphatase, which is phosphorylated by Ca2+/CaM-dependent protein kinase II (CaM-kinase II) has been identified. Analyses of 32P release from tryptic and cyanogen bromide peptides derived from [32P]calcineurin plus direct sequence determination established the site as -Arg-Val-Phe-Ser(PO4)-Val-Leu-Arg-, which conformed to the consensus phosphorylation sequence for CaM-kinase II (Arg-X-X-Ser/Thr-). This phosphorylation site is located at the C-terminal boundary of the putative CaM binding domain in calcinerin (Kincaid, R. L., Nightingale, M. S., and Martin, B. M. (1988) Proc. Natl. Acad. Sci. U. S. A. 85, 8983-8987), thereby accounting for the observed inhibition of this phosphorylation when Ca2+/CaM is bound to calcineurin. Since the phosphorylation site sequence also contains elements of the specificity determinants for Ca2+/phospholipid-dependent protein kinase (protein kinase C) (basic residues both N-terminal and C-terminal to Ser/Thr), we tested calcineurin as a substrate for protein kinase C. Protein kinase C catalyzed rapid stoichiometric phosphorylation, and the characteristics of the reaction were the same as with CaM-kinase II: 1) the phosphorylation was blocked by binding of Ca2+/CaM to calcineurin; 2) phosphorylation partially inactivated calcineurin by increasing the Km (from 9.9 +/- 1.1 to 17.5 +/- 1.1 microM 32P labeled myosin light chain); and 3) [32P]calcineurin exhibited very slow autodephosphorylation but was rapidly dephosphorylated by protein phosphatase IIA. Tryptic and thermolytic 32P-peptide mapping and sequential phosphoamino acid sequence analysis confirmed that protein kinase C and CaM-kinase II phosphorylated the same site. PMID- 2550448 TI - Purification of A1 adenosine receptor from rat brain membranes. AB - The A1 adenosine receptor from rat brain membranes has been purified about 50,000 fold to apparent homogeneity by sequential use of affinity chromatography on immobilized xanthine amine congener-agarose, hydroxylapatite chromatography, and reaffinity chromatography. The overall yield starting from the membranes was approximately 4%. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified preparation gave a broad single band of an apparent molecular weight of 34,000 either by silver staining or autoradiogram after radioiodination. The purified receptor bound approximately 24 nmol of 8-cyclopentyl-1,3 [3H]dipropylxanthine/mg of protein with a dissociation constant of 1.4 nM. This maximum specific binding value is consistent with the expected theoretical specific activity (29.4 nmol/mg) for a protein with a molecular mass of 34,000 daltons if it is assumed that there is one ligand-binding site/receptor molecule. Affinity-labeling experiments using [3H]p-phenylenediisothiocyanate-xanthine amine congener showed that the Mr = 34,000 protein band contained the ligand binding sites. The purified receptor gave a typical A1 adenosine receptor pharmacological specificity similar to that of unpurified receptor preparations. PMID- 2550449 TI - A new class of calcium entry blockers defined by 1,3-diphosphonates. Interactions of SR-7037 (belfosdil) with receptors for calcium channel ligands. AB - Tetrabutyl-2(2-phenoxyethyl)-1,3-propylidene diphosphonate (SR-7037) completely displaced dihydropyridine [( 3H]PN200-110), phenylalkylamine [( 3H]D888), and benzothiazepine [( 3H]diltiazem) ligands from brain L-type calcium channels. Half maximal inhibition of [3H]PN200-110 binding occurred at 19 nM with a Hill coefficient of 0.96. SR-7037 primarily decreased the affinity for [3H]PN200-110 with a small, but significantly, effect on the maximal binding capacity. Kinetic studies showed that this was due to an increased radioligand dissociation rate from 0.04 min-1 to 0.43 min-1 in the presence of the diphosphonate. Displacement of [3H]D888 by SR-7037 was biphasic with respective IC50 of 44 and 8400 nM. Likewise, unlabeled (-)-D888 identified two sites with IC50 values of 0.9 and 27 nM. Both SR-7037 (1000 nM) and D888 (200 nM) accelerated radioligand dissociation about 2-fold. [3H]Diltiazem binding was inhibited by SR-7037 with an IC50 value of 29 nM. The inhibition of dihydropyridine binding by SR-7037 is enhanced by most divalent cations at millimolar concentrations with the following potency: Mn2+ greater than Mg2+ greater than Ca2+ greater than Co2+. Barium has the opposite effect. The half-maximal effect of calcium occurred at 6 microM free ion. Specific binding of [3H]D888 was antagonized in the presence of 1 mM CaCl2. It is concluded that SR-7037 has allosteric interactions with the dihydropyridine receptor of the L-type calcium channel. The differential effect of Ca2+ on the potency of D888 and diltiazem relative to that of SR-7037 indicates that the three drugs may bind to nonequivalent sites. These results support specific calcium channel inhibition, possibly at a novel site, as the primary mechanism of the diphosphonate's pharmacological actions. PMID- 2550450 TI - The elastin receptor shows structural and functional similarities to the 67-kDa tumor cell laminin receptor. AB - Laminin- and elastin-binding proteins were isolated by ligand affinity chromatography from plasma membranes of fetal bovine auricular chondroblasts and human A2058 melanoma cells. From both cell types, a 67-kDa protein was identified which bound to either elastin or laminin affinity resins. Structural and functional similarities between the elastin and laminin-binding proteins were suggested by 1) cross-reactivity between antibodies directed against the two proteins; 2) elution of the laminin receptor from laminin columns with soluble elastin peptides; and 3) modulation of substrate binding by galactoside sugars. In addition, extraction properties indicate that both receptors are peripheral membrane proteins whose association with the cell surface is mediated by their lectin properties. Mapping of the binding site on laminin suggests that the 67 kDa chondroblast receptor interacts with a hydrophobic elastin-like sequence in domain V of the B1 chain, and chemotaxis studies indicate that cell migration to elastin peptides and laminin involves the same receptor. PMID- 2550451 TI - Virus infection and interferon can activate gene expression through a single synthetic element, but endogenous genes show distinct regulation. AB - Virus inducible elements (IE) in promoters of mouse alpha-interferon and human beta 1-interferon genes contain multiple copies of the hexanucleotide sequence AGT-GAA or its variants which are also found in the interferon-stimulated response element of genes transcriptionally induced by interferon. We have examined the similarities between virus and interferon induction of gene expression and the role of AGTGAA and AAT-GAA hexamers in these responses. Hybrid plasmids were constructed by inserting the IE region, the alpha 4 promoter, or the multiple copies of AGTGAA or AAT-GAA 5' to the inactive-45 human immunodeficiency-chloramphenicol acetyltransferase hybrid gene, and their inducible expression was studied in a transient expression assay. In L-cells, multiple hexamers were efficiently induced both by infection with Newcastle disease virus and by interferon treatment; while the alpha 4 promoter and the IE inducible region were induced predominantly by virus rather than by interferon. In order to dissociate the effect of virus and endogenous interferon on the induction process, we examined the gene expression in Vero cells, which have undergone homozygous deletion of type 1 interferon genes, and in VNPT-159 cells, which were derived from Vero cells by insertion of an inducible human interferon beta 1 gene. The results show that while the alpha 4 promoter was efficiently induced only by virus in both cell types, the constructs containing shorter segments of the IE were induced by both virus and interferon in Vero cells. However, the inducibility by interferon was not detected in VNPT-159 cells, suggesting that the presence of endogenous interferon suppresses interferon induced expression of hexanucleotide repeats and the short inducible region. In contrast, virus inducibility of endogenous interferon-stimulated genes, ISG-15 and ISG-54, was about 100-fold more efficient in VNPT-159 cells than in Vero cells, suggesting that this induction is largely mediated through synthesis of endogenous interferon. Hence, endogenous interferon may play a role in the autoregulation of both interferon genes and interferon-stimulated genes. PMID- 2550452 TI - Functional characterization of cAMP-binding mutations in type I protein kinase. AB - A mutant form of the type I regulatory subunit (RI) of cAMP-dependent protein kinase has been cloned and sequenced (Clegg, C. H., Correll, L. A., Cadd, G. C., and McKnight, G. S. (1987) J. Biol. Chem. 262, 13111-13119) which contains two point mutations in the site B cAMP-binding site, a Gly to Asp at position this report, the effect of each independent mutation on the rate of dissociation of cAMP from RI, the cAMP-mediated activation of holoenzyme and the inducibility of cAMP-responsive genes has been characterized. Dissociation of cAMP from either recombinant wild type RI or the B1 mutant demonstrated biphasic kinetics, indicating two sites with different affinities for cAMP. Dissociation from the B2 subunit, however, was monophasic and very rapid indicating that site B had been destroyed and that the rate of dissociation from site A was increased. The cAMP activation constants (Ka) of the wild type and B1 holoenzymes were 40 and 188 nM, respectively, and demonstrated positive cooperativity, with Hill coefficients of 1.61 for the wild type and 1.67 for B1. The B2 holoenzyme required much greater concentrations of cAMP, 4.7 microM, for half-maximal activation and did not display positive cooperativity. Constitutive expression in mouse AtT20 pituitary cells of the B1 mutant resulted in only a small shift in the Ka for kinase activation in these cells compared with B2 expression which increased the Ka by more than 100-fold. Transient expression of the B1 subunit in human JEG-3 choriocarcinoma cells inhibited forskolin activation of a cAMP-responsive promoter by 35% whereas similar expression of the B2 RI subunit inhibited the response by 90%. These results suggest that the Gly to Asp mutation at amino acid 324 completely blocks cAMP binding to site B whereas the Arg to His mutation at position 332 causes a more subtle alteration in cAMP binding. Expression of either mutant RI in animal cells results in a dominant repression of cAMP dependent protein kinase activity and cAMP-dependent protein kinase-mediated processes. PMID- 2550453 TI - An antibody-induced enhancement of the transducin-stimulated cyclic GMP phosphodiesterase activity. AB - In this work we have characterized the ability of a carboxyl peptide-specific antibody (AS/7), raised against the alpha subunit of transducin (alpha T), to potentiate the stimulation of the cyclic GMP phosphodiesterase (PDE) by transducin. The complexation of the purified guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S)-bound form of alpha T (alpha T.GTP gamma S) with AS/7 results in a 2-5-fold enhancement in the total levels of cyclic GMP hydrolysis measured after 1 min. This potentiation by AS/7 cannot be attributed simply to an increase in the apparent affinity of alpha T.GTP gamma S for the effector enzyme, nor to an increased affinity of the enzyme for the substrate cyclic GMP. The AS/7-induced potentiation is specific for alpha T.GTP gamma S-PDE interactions; this antibody has no effect on the activity of the trypsin-activated PDE nor on the ability of the GDP-bound form of alpha T to inhibit the trypsin-activated enzyme (Kroll, S., Phillips, W. J., and Cerione, R. A. (1989) J. Biol. Chem. 264, 4490-4497). Phosphatidylcholine vesicles also will enhance the alpha T.GTP gamma S-stimulated PDE activity (1.5-2-fold) relative to that measured in the absence of a lipid milieu. However, the potentiations of alpha T-stimulated cyclic GMP hydrolysis elicited by AS/7 and lipids represent separate events. Titration profiles describing the AS/7-induced potentiation, as a function of the amount of antibody added to the assay mixtures, indicate that maximal activity occurs when there is one molecule of AS/7 per two molecules of alpha T.GTP gamma S; the AS/7-induced potentiation is lost when AS/7 much greater than alpha T. GTP gamma S, i.e. conditions which favor the formation of monovalent AS/7-alpha T.GTP gamma S complexes. When the AS/7 is papain-treated to yield monovalent antibody molecules, complexation between these monovalent antibodies and alpha T still occurs (as reflected by the ability of these antibodies to block rhodopsin-alpha T coupling); however, the potentiation of the alpha T.GTP gamma S-stimulated PDE activity is lost. Taken together, these results suggest that the AS/7-induced potentiation of alpha T-stimulated activity is dependent on the bivalent nature of the antibody, and maximal stimulation of PDE activity is achieved by the interactions of two activated-alpha T molecules with a single molecule of PDE. PMID- 2550454 TI - Subcellular distribution of "peripheral type" binding sites for [3H]Ro5-4864 in guinea pig lung. Localization to the mitochondrial inner membrane. AB - Binding of [3H]Ro5-4864, a specific ligand for "peripheral type" benzodiazepine receptors, was determined in subcellular fractions of guinea pig lung. Even though the level of binding was predominant in the mitochondrial fraction, nuclear and cytosolic fractions also contained significantly measurable amounts of binding sites. The presence of binding sites in the microsomal fraction and in a fraction intermediate in density between the mitochondria and microsomes depended on which buffer was used to homogenize the tissue. If calcium-containing mannitol buffer was used, binding was negligible in the postmitochondrial organelles. However, in the case of sucrose buffer which did not contain any calcium, the postmitochondrial organelle fractions contained measurable amounts of binding sites. Most probably, these binding sites were of mitochondrial and nuclear origin. Furthermore, binding sites in the mitochondria were associated with the succinic dehydrogenase-enriched mitochondrial inner membrane, but not with the monoamine oxidase- and cholinephosphotransferase-enriched outer mitochondrial membrane. Furthermore, several proteolytic enzymes caused a decrease in binding of the ligand to the mitochondrial membrane only under hypotonic conditions and not under isotonic conditions, suggesting that the location of the receptors is inside the mitochondria. PMID- 2550455 TI - Developmental expression of the tissue insulin-like growth factor II/mannose 6 phosphate receptor in the rat. Measurement by quantitative immunoblotting. AB - We used quantitative immunoblotting to measure the total tissue insulin-like growth factor II/mannose 6-phosphate (IGF-II/Man-6-P) receptor in the rat. Whole embryos (6-15 days of gestation) and tissues from 16- and 20-day-old fetal and 5 , 10-, 20-, and 40-day-old postnatal rats were placed in liquid nitrogen, extracted with 2% Triton X-100, and boiled in 2% sodium dodecyl sulfate. The extracts were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis together with aliquots of a highly purified rat IGF-II/Man-6-P receptor standard. The protein bands were transferred from the gel to nitrocellulose sheets by electroelution. The nitrocellulose sheets were incubated with a specific IGF-II/Man-6-P receptor antiserum (3637). The immunoblots were developed with 125I-protein A and an immunoperoxidase stain. Stained areas were cut from the immunoblots, and radioactivity was measured in a gamma-counter. IGF II/Man-6-P receptor levels were high in fetal tissues and in most tissues declined dramatically in late gestation and/or in the early postnatal period. Concentrations in 16-day-old fetal tissues, expressed as percent of total protein in the extract, were: heart, 1.7; placenta, 1.0; lung, 0.7; intestine, 0.7; muscle, 0.5; kidney, 0.5; liver, 0.3; and brain, 0.1. In whole embryos (6-15 days of gestation), the IGF-II/Man-6-P receptor ranged between 0.1 and 0.4% of total protein in the extract. The IGF-II/Man-6-P receptor size varied within approximately 20 kDa among different tissues and also varied with developmental age in the same tissue. We conclude that the IGF-II/Man-6-P receptor is a major cellular protein in some fetal tissues and that the developmental pattern of receptor expression suggests that the receptor plays an important role in fetal growth and development. PMID- 2550456 TI - Recognition of modified forms of ribonuclease A by the ubiquitin system. AB - The substrate specificity of the ubiquitin (Ub) conjugation system was explored with regard to recognition of unfolded conformation and/or oxidized methionine residues in six derivatives of bovine RNase A. Based on the following observations, ubiquitination of RNase A substrates by the enzymes in a rabbit reticulocyte extract appears to correlate with unfolded conformation rather than with methionine oxidation. 1) Methionine oxidation in already unfolded forms of RNase A does not enhance ubiquitination. 2) Fluorescence measurements and iodoacetate trapping of free sulfhydryls show that the disulfide bonds of MetSO RNase A, in which the 4 methionine residues are oxidized to the sulfoxide, are reduced by 2 mM dithiothreitol (DTT) in standard Ub conjugation assays so that this derivative also is unfolded. 3) Although MetSO-RNase A is ubiquitinated in the absence of DTT, its intrinsic fluorescence, cation-exchange properties, and susceptibility to reduction indicate a non-native conformation. 4) Methionine sulfoxide-containing peptides that mimic regions of RNase A fail to inhibit conjugation of 125I-Ub to MetSO-RNase A. Ub adducts to two of the six derivatives (MetSO- and reduced/carboxamidomethylated MetSO-RNase A) increase when DTT is omitted from the reactions. Ubaldehyde, an inhibitor of isopeptidases that disassemble Ub-protein conjugates, increased product yields and reduced or abolished the DTT effect, suggesting that an isopeptidase specific for these two RNase A derivatives may be inactivated by oxidation. Ub conjugates of the other RNase A derivatives also increase with Ub-aldehyde but are unaffected by DTT. PMID- 2550457 TI - Structure of testicular angiotensin-converting enzyme. A segmental mosaic isozyme. AB - The complete amino acid sequence of rabbit testicular angiotensin-converting enzyme has been deduced from the sequence of the corresponding cDNA clone. A protein of the expected molecular weight of 84,000 was translated in vitro from the mRNA encoded by this cDNA. All of the previously determined sequences of seven tryptic peptides from the enzyme are present in the deduced sequence, thus confirming the identity of the protein. From the deduced sequence it appears that the protein contains a signal peptide at the amino terminus and a hydrophobic anchoring domain near the carboxyl terminus. Northern analysis with oligonucleotide probes, whose sequences represented different regions of the cDNA, revealed not only the regions of extensive homology between the mRNAs encoding the testicular and the pulmonary isozymes but also a stretch of sequence near the 5' end unique to the testicular mRNA. PMID- 2550458 TI - Studies of the regulatory mechanism of Ca2+/calmodulin-dependent protein kinase II. Mutation of threonine 286 to alanine and aspartate. AB - A cDNA clone for the alpha subunit of mouse brain Ca2+/CaM-dependent protein kinase II (CaM-kinase II) was transcribed in vitro and translated in a rabbit reticulocyte lysate system. Inclusion of [35S]methionine in the translation system yielded a single 35S-polypeptide of about 50 kDa. When the translation system was assayed for CaM-kinase II activity, there was a 5-10-fold enrichment of kinase activity which was totally dependent on Ca2+/calmodulin (CaM). Both the 50-kDa 35S-polypeptide and the Ca2+/CaM-dependent protein kinase activity were quantitatively immunoprecipitated by rat brain CaM-kinase II antibody. When the translated wild-type kinase was subjected to autophosphorylation conditions in the presence of Ca2+, CaM, Mg2+, and ATP, the Ca2+-independent activity (assayed in the presence of [ethylenebis(oxyethylenenitrilo)]tetraacetic acid) increased from 5.8 +/- 0.7 to 26.5 +/- 2.1% of total activity (assayed in the presence of Ca2+/CaM). These properties confirm the identity of the kinase translated in vitro as CaM-kinase II. The role of Thr-286 autophosphorylation in formation of the Ca2+-independent activity was investigated by site-directed mutation of Thr 286 to Ala (Ala-286 kinase) and to Asp (Asp-286 kinase). The Ala-286 kinase was completely dependent on Ca2+/CaM for activity prior and subsequent to autophosphorylation. The Asp-286 kinase exhibited 21.9 +/- 0.8% Ca2+-independent activity, and this was not increased by autophosphorylation. These results establish that introduction of negative charge(s) at residue 286, either by autophosphorylation of Thr or by mutation to Asp, is sufficient and necessary to generate the partially Ca2+-independent form of CaM-kinase II. PMID- 2550459 TI - Annealing of gelsolin-severed actin fragments by tropomyosin in the presence of Ca2+. Potentiation of the annealing process by caldesmon. AB - Previous results from our laboratory have shown that 1) cultured rat cells contain two classes of tropomyosin (TM), one (high Mr TMs) with higher Mr values and greater affinity for actin than the other (low Mr TMs); 2) presaturation of F actin with high Mr TMs, but not with low Mr TMs, inhibits both actin-severing and actin binding activities of gelsolin; and 3) nonmuscle caldesmon not only enhances the inhibitory effects of high Mr TMs but also makes low Mr TMs capable of inhibiting the severing activity of gelsolin (Ishikawa, R., Yamashiro, S., and Matsumura, F. (1989) J. Biol. Chem. 264, 7490-7497). These results suggest that gelsolin has much lower affinity for F-actin-TM-caldesmon complexes than for pure F-actin. We have therefore examined whether addition of TM and/or caldesmon to gelsolin-severed actin filaments can make gelsolin dissociate from barbed ends of actin filaments, resulting in annealing of short actin filaments into long ones. Flow birefringence and electron microscopic studies have suggested that high Mr TMs slowly and partially anneal gelsolin-severed actin fragments in 3 h, whereas low Mr TMs have no effects. Nonmuscle caldesmon greatly potentiates the effects of high Mr TMs and accelerates the process to 20 min, whereas nonmuscle caldesmon alone shows no effects. Furthermore, nonmuscle caldesmon makes low Mr TMs capable of reversing gelsolin-severing action. Actin binding assay has shown that gelsolin (or a gelsolin-actin complex) is dissociated from these annealed actin filaments. Smooth muscle TM and smooth muscle caldesmon also appear to anneal gelsolin-severed actin fragments as do high Mr TMs and nonmuscle caldesmon. Calmodulin decreases the potentiation effects of caldesmon as calmodulin inhibits actin binding of caldesmon. These results suggest that tropomyosin and caldesmon may regulate both capping and severing activities of gelsolin. PMID- 2550460 TI - The ryanodine receptor-Ca2+ release channel complex of skeletal muscle sarcoplasmic reticulum. Evidence for a cooperatively coupled, negatively charged homotetramer. AB - The subunit structure of the rabbit skeletal muscle ryanodine receptor-Ca2+ release channel complex was examined following solubilization of heavy sarcoplasmic reticulum membranes in two zwitterionic detergents, 3-[(3 cholamidopropyl)dimethylammonio]-1-propanesulfonic acid (Chaps) and Zwittergent 3 14. High and low affinity [3H]ryanodine binding was retained upon solubilization of the complex in Chaps but was lost in Zwittergent 3-14. The purified complex migrated as a single peak with an apparent sedimentation coefficient of approximately 30 and approximately 9 S upon density gradient centrifugation and with isoelectric points of 3.7 and 3.9 upon two-dimensional gel electrophoresis in Chaps and Zwittergent 3-14, respectively. Electron microscopy of negatively stained samples indicated that the distinct four-leaf clover structure of the ryanodine receptor observed in Chaps disappeared following Zwittergent treatment of the 30 S complex and instead showed smaller, round particles. Ferguson plot analysis following sodium dodecyl sulfate-polyacrylamide gel electrophoresis of partial and fully cross-linked and incompletely denatured complexes suggested a stoichiometry of four Mr approximately 400,000 peptides/30 S ryanodine receptor oligomer. [3H]Ryanodine binding to the membrane-bound receptor in 50 microM--1 mM free Ca2+ revealed the presence of both high affinity (KD = 8 nM, Hill coefficient (nH) = 0.9) and low affinity (nH approximately 0.45) sites with a ratio of 1:3. Reduction in free Ca2+ to less than or equal to 0.1 microM or trypsin digestion of the membranes resulted in loss of high affinity but not low affinity ryanodine binding (Hill KD = 5,000 nM, nH = 0.9). Addition of 20 mM caffeine to the nanomolar Ca2+ medium decreased the Hill KD to 1,000 nM without changing the Hill coefficient. Occupation of the low affinity sites altered the rate of [3H]ryanodine dissociation from the high affinity sites. Single channel recordings of the purified ryanodine receptor channel incorporated into planar lipid bilayers also indicated the existence of high and low affinity sites for ryanodine, occupation of which resulted in formation of a subconducting and completely closed state of the channel, respectively. These results are compatible with a subunit structural model of the 30 S ryanodine receptor-Ca2+ release channel complex which comprises a homotetramer of negatively charged and allosterically coupled polypeptides of Mr approximately 400,000. PMID- 2550461 TI - Molecular cloning and characterization of a chromosomal gene for human eosinophil peroxidase. AB - Using human myeloperoxidase cDNA as a probe, a chromosomal gene related to myeloperoxidase was isolated from a human gene library. Comparison of the amino acid sequence deduced from the nucleotide sequence of the cloned gene with that of human eosinophil peroxidase purified from buffy coats has indicated that the isolated gene is the chromosomal gene for human eosinophil peroxidase. Like human myeloperoxidase gene, human eosinophil peroxidase gene consists of 12 exons and 11 introns spanning about 12 kilobases. The gene can code for a protein of 715 amino acids with a calculated Mr of 81,036. The heavy chain and the light chain of eosinophil peroxidase were located on the COOH and NH2 terminus of the protein, respectively. The coding sequences of eosinophil peroxidase and myeloperoxidase show homologies of 72.4% at the nucleotide and 69.8% at the amino acid level, while little homology was found in the 5'-flanking region. Northern hybridization and S1 mapping analysis of RNA from human leukemic cells have indicated that the eosinophil peroxidase gene is expressed in the eosinophilic subline of human HL-60 cells but not in the neutrophilic subline or in parental HL-60 cells. PMID- 2550462 TI - Cloning and sequencing of the cDNA for a 13th different subunit (IHQ) of beef heart cytochrome c oxidase. AB - The mammalian cytochrome c oxidase is a complex of 13 different subunits. We present the full amino acid sequence of the one remaining uncharacterized subunit, subunit IHQ in the nomenclature used here, VIIb in the numbering system of Kadenbach and colleagues (e.g. Kadenbach, B., and Merle, P. (1981) FEBS Lett. 135, 1-11). A partial protein sequence was obtained from the purified subunit isolated by gel filtration procedures. This information was used to synthesize an oligonucleotide probe which was then used to isolate a cDNA clone encoding the subunit. This cDNA for subunit IHQ is 480 base pairs long and encodes a polypeptide which is either 83 or 88 amino acids long, including an N-terminal leader sequence of either 27 or 32 residues. The molecular weight of the mature subunit IHQ is 6350 based on the amino acid sequence deduced from the gene. The leader sequence is typical of other mitochondrial target sequences in having several positively charged residues but no negatively charged side chains. PMID- 2550463 TI - Repression of polyoma virus DNA replication by 5'-flanking region of mouse DNA polymerase beta gene containing transcriptional silencer elements. AB - Dual cis-acting silencer elements are located upstream of the mouse DNA polymerase beta gene (Yamaguchi, M., Hayashi, Y., and Matsukage, A. (1989) J. Biochem. (Tokyo) 105, 79-83). In order to examine possible involvement of transcriptional silencer elements in the regulation of DNA replication, we have utilized a transient replication system of the plasmid DNA carrying replication origin of polyoma virus DNA in mouse MOP8 cells, which is constitutively producing polyoma virus large T-antigen. The polyoma virus origin of DNA replication is composed of three cis-acting genetic elements called alpha, beta, and core, in which alpha and beta elements correspond to enhancer domains. When the 5'-flanking regions of the DNA polymerase beta gene containing silencer elements were placed at the late gene border of alpha element, they effectively repressed the DNA replication. However, when placed at the early gene border of core element, it only marginally repressed the DNA replication. These results suggest that the silencer elements at cis position repress polyoma virus DNA replication by impeding the enhancer function that activates the DNA replication. PMID- 2550464 TI - Inhibitor analysis of synapsis by resolvase. AB - Previously, we isolated several inhibitors that block the site-specific recombination reaction mediated by the Tn3-encoded resolvase protein. One class of inhibitors blocks resolvase binding to the recombination (res) sitc, and a second class inhibits synapse formation between resolvase and two directly repeated res sites. In this report, we identify an inhibitor, A20832, that does not inhibit resolvase binding to res, as measured by filter binding, or synapse formation. Inhibition of resolvase-promoted site-specific recombination by A20832 occurs postsynaptically at strand cleavage. DNase I analysis in the presence of A20832 indicates that only site I of res is bound by resolvase. PMID- 2550465 TI - Degradation of apolipoprotein II mRNA occurs via endonucleolytic cleavage at 5' AAU-3'/5'-UAA-3' elements in single-stranded loop domains of the 3'-noncoding region. AB - Degradation intermediates of the estrogen-regulated apolipoprotein (apo) II mRNA were identified by S1 nuclease mapping and primer extension analysis. S1 mapping of poly(A)-RNA detected a series of mRNAs truncated at specific sites in the 3' noncoding region. Many of these sites were also detected by primer extension analysis indicating that truncated molecules resulted from endonucleolytic cleavage in the 3'-noncoding region. Identical cleavage sites were seen with RNA from estrogen-treated animals or from animals withdrawn from hormone under conditions where apoII mRNA degraded in the slow (t1/2 = 13 h) or rapid (t1/2 = 1.5 h) decay mode. No differences were seen in poly(A) tail length or heterogeneity among these conditions. These results indicate that the estrogen induced alteration in apoII mRNA turnover does not involve a new pathway of degradation, but, more likely, involves an increased targeting of the mRNA for degradation by a preexisting pathway. These data are consistent with a mechanism in which the initial step in apoII mRNA degradation is an endonucleolytic cleavage in the 3'-noncoding region without prior removal of the poly(A) tail. The endonucleolytic cleavage sites occurred predominantly at 5'-AAU-3' or 5'-UAA 3' trinucleotides found in single-stranded domains in a secondary structure model of the naked mRNA (Hwang, S-P. L., Eisenberg, M., Binder, R., Shelness, G. S., and Williams, D. L. (1989) J. Biol. Chem. 264, 8410-8418). The structure of the 3'-noncoding region in polyribosomal messenger ribonucleoprotein was examined by titrations of liver homogenates with dimethyl sulfate and cobra venom RNase. The results suggest that the typical cleavage site is a 5'-AAU-3' or 5'-UAA-3' trinucleotide in an accessible single-stranded loop domain. Single-stranded domains alone or accessible domains alone are not sufficient for cleavage. Similarly, 5'-AAU-3' or 5'-UAA-3' trinucleotides alone are not sufficient for cleavage. Localization of these trinucleotides to accessible single-stranded domains in the polyribosomal messenger ribonucleoprotein may provide the specificity for cleavage during targeted degradation. PMID- 2550466 TI - Cyclic AMP-dependent protein kinase in canine pancreatic rough endoplasmic reticulum. AB - A canine pancreas homogenate was subfractionated by several differential centrifugation steps. The distribution of cAMP-dependent protein kinase in the various fractions was monitored by assaying [3H]cAMP binding and photo-cross linking of the regulatory subunits of the enzyme (RI and RII) with radiolabeled 8 azido-cAMP. The distribution of the kinase was also compared to that of markers for the plasma membrane, the endoplasmic reticulum and the cytosol. While our results confirm previous studies suggesting the presence of cyclic AMP-dependent protein kinase in the cytosol and Golgi, a significant amount of the total [3H] cAMP binding and photolabeled R subunits (both RI and RII) were found in rough microsomes (RM). The association is relatively resistant to extraction with EDTA, low and high ionic strength solutions. These extractions unmasked several new phosphorylation substrates in the "stripped" RM that were inaccessible in the RM, possibly because they were covered by ribosomes or peripheral membrane proteins. RII with a molecular mass of 52 kDa (RII-52 kDa) was the predominant RII found in the cytosolic fraction, whereas RII-52 kDa and RII with a molecular mass of 54 kDa (RII-54 kDa) were approximately equally enriched in the RM fraction. The mobility of the RII-52 kDa-photolabeled band could be shifted to the mobility of the RII-54 kDa band by phosphorylation with purified catalytic subunit and ATP, indicating that they represent "dephospho" and "phospho" forms of RII, respectively. A more precise localization to the rough endoplasmic reticulum was accomplished by isopycnic floatation in sucrose gradients. The enzyme cobanded at the density of rough microsomes and shifted to the lower density of "stripped" microsomes after treatment with puromycin/high salt, which specifically removes ribosomes. PMID- 2550467 TI - The effect of gamma irradiation on injectable human amnion collagen. AB - The effect of gamma irradiation on the physicochemical properties of injectable human amnion collagen was investigated. Pepsin-extracted human amnion collagen was purified, reconstituted, and irradiated with varying doses of gamma irradiation (0.25 Mrads to 2.5 Mrads). Gamma irradiation had a significant impact on the physical characteristics of the collagen. The neutral solubility of collagen in PBS at 45 degrees C was decreased from 100% for the nonirradiated control sample to 16% for the 2.5 Mrads irradiated sample. SDS polyacrylamide gel electrophoresis also demonstrated the dose-dependent effect of gamma irradiation on collagen cross-links. Electron microscopic observation revealed that even at low irradiation dose (0.25 Mrads), collagen fibril diameter increased. The average diameter was 50 nm for nonirradiated control fibrils, while 4.4 percent of the irradiated collagen fibrils had a diameter greater than 100 nm. Irradiated collagen showed little evidence of damage. Well-preserved cross-striations were found in collagen fibrils at all doses of irradiation. Native amnion collagen irradiated with gamma rays demonstrated a slight increase in resistance to collagenase degradation compared with nonirradiated native collagen samples. Increased resistance to collagenase did not correlate with increasing irradiation dose. After 30 min of incubation at 37 degrees C, both irradiated and nonirradiated collagen was completely digested by collagenase. However, gamma irradiated collagen did become more sensitive to hydrolysis by trypsin. The higher the irradiation doses used, the greater sensitivity to trypsin was observed. At 0.25 Mrads irradiation only a slight increase was found. No marked differences in amino acid composition were noted among the high dose irradiated, low dose irradiated and control amnion collagen. PMID- 2550468 TI - Behavior of platelets and leukocytes on the luminal surface of small caliber polyurethane grafts. AB - With scanning electron microscopy, we examined the behavior of platelets and leukocytes on the luminal surface of small caliber polyurethane grafts implanted into small arteries of dogs. Thirty-five grafts were implanted to the carotid and/or femoral arteries. The animals were treated with aspirin and/or DN9693, an inhibitor of phosphodiesterase. In the group treated with aspirin (40 mg/kg i.v.), the deposition of platelets on the luminal surface of the implanted polyurethane grafts was suppressed and the luminal surface was covered with adherent leukocytes. Fibrin nets were formed on the adherent leukocytes. In addition, the adhesion of leukocytes on the grafts was considerably suppressed in the group treated with DN9693 (50 micrograms/kg/min i.v.), and the formation of fibrin nets was markedly reduced. In contrast, in the control group the luminal surface was covered with numerous platelets and some leukocytes, which formed thrombi. These findings suggest that leukocytes adhere primarily to the prosthetic graft and play an important role in the initiation of fibrin formation. PMID- 2550469 TI - Perioperative alteration of neutrophil functions after open heart and other major surgeries. A comparative study. AB - The prolonged effects of cardiopulmonary bypass on neutrophil functions, as indicated by the capacity to produce superoxide and leukotrienes, were examined on the first postoperative day in ten patients who underwent open heart surgery and ten who underwent other major surgery. The superoxide and leukotriene C4 producing capacities of the neutrophils were significantly decreased postoperatively in both groups. The leukotriene B4 producing capacity, by contrast, was significantly increased postoperatively. However, there was no significant difference in values between the two groups in the pre- and postoperative periods. In both groups, neutrophil functions were significantly altered on the first postoperative day, but no specific alteration due to open heart surgery was observed. PMID- 2550470 TI - Protein kinase C and a viral K-RAS protein cooperatively enhance the response of adenylate cyclase to stimulators. AB - The protein kinase C stimulator TPA (12-O-tetradecanoyl phorbol-13-acetate) enhanced the responsiveness of adenylate cyclase to IPR (isoproterenol) and PGE1 (prostaglandin E1) in quiescent tsKSV-NRK cells at the nonpermissive 41 degrees C. Reactivating the thermolabile mitogenic/oncogenic K-ras protein in tsKSV-NRK cells by dropping the temperature to 36 degrees C also enhanced the responsiveness of adenylate cyclase to IPR and PGE1. The enhancement was transient and peaked at 6 hours after the temperature shift. This enhanced responsiveness was specifically due to the reactivated viral K-ras protein rather than the temperature shift because the same temperature shift did not affect adenylate cyclase responsiveness in uninfected NRK cells, nor was it a result of the mitogenic stimulus since reacting the mitogenic pp60v-src protein in tsASV NRK cells did not affect adenylate cyclase responsiveness. The increased responsiveness of adenylate cyclase at 6 hours after the temperature shift was not a result of elevated membrane-associated PKC activity. However, the reactivated viral K-ras protein strongly increased the stimulability of membrane associated PKC by TPA and it further increased TPA's ability to enhance the responsiveness of adenylate cyclase to IPR and PGE1. Thus, a viral K-ras protein and membrane-associated protein kinase C can cooperate to increase the responsiveness of adenylate cyclase to agonists. PMID- 2550471 TI - Effect of pentoxifylline on developmental changes in neutrophil cell surface mobility and membrane fluidity. AB - Polymorphonuclear leukocytes (PMNs) from human neonates respond less efficiently to chemotactic factor stimulation than do PMNs from adults. The biologic mechanisms underlying this developmental process are poorly understood. In previous studies, we have found that pentoxifylline, an agent report to enhance membrane deformability, increased the chemotactic response of neonatal PMNs. In the present studies, we have examined the effect of pentoxifylline on cell surface mobility and membrane fluidity by assessing fluorescent concanavalin A (Con A) capping and fluorescent polarization (FP). Baseline Con A capping was lower in the PMNs of neonates when compared to PMNs from adult controls. Colchicine, which increases capping by disrupting microtubules, exaggerated the differences between the adult and neonatal PMNs. Following exposure of neonatal PMNs to pentoxifylline, colchicine enhanced Con A capping to levels equivalent to those of colchicine-treated PMNs from adults. Employing a fluorescence polarization (FP) assay, we found the fluid state of the membrane of PMNs from neonates was significantly less than that of adult controls. Pentoxifylline alone significantly increased the fluidity of the cell membranes of neonatal PMNs while decreasing elevated basal levels of F-actin in the cell. These data suggest an intrinsic cytoskeletal difference in the PMNs of neonates that may be responsive to pharmacologic manipulation. PMID- 2550472 TI - Diacylglycerol, but not inositol 1,4,5-trisphosphate, accounts for platelet derived growth factor-stimulated proliferation of BALB 3T3 cells. AB - Recently we found that an intracellular event related to phosphatidylinositol 4,5 bisphosphate (PIP2) is crucial for platelet-derived growth factor (PDGF)-induced mitogenesis in fibroblastic cells (Matuoka, K., et al.: Science 239:640-643, 1988). In the present study we examined the mitogenic effects of PIP2 and its hydrolysis products introduced into the cytoplasm of BALB 3T3 cells by micro injection to confirm the role of PIP2 hydrolysis in PDGF stimulation of cell proliferation. Injection of 1,2-dioleylglycerol (diolein) into serum-deprived quiescent cells induced DNA synthesis with the same time course as that induced by exposure of the cells to PDGF and, in the presence of PDGF, caused no additional increase in the cell population entering S phase. The injection of PIP2, inositol 1,4,5-trisphosphate, or 1,2-dioleylphosphatidic acid into the cells did not induce mitogenesis. Consistent results were obtained in experiments in which the cells were exposed to 1-oleyl-2-acetylglycerol (OAG) and ionomycin; namely, OAG stimulated proliferation of BALB 3T3 cells, but ionomycin did not induce any mitogenesis. Desensitization of the protein kinase C pathway by prolonged exposure of the cells to phorbol ester abolished the induction of cell proliferation by subsequent injection of diolein or exposure to phorbol ester or OAG as well as by PDGF challenge. These findings strongly suggest that activation of the protein kinase C system following formation of diacylglycerol by PIP2 hydrolysis is mainly responsible for the mitogenic action of PDGF on BALB 3T3 cells. PMID- 2550474 TI - Oxygen gradients in CHO cells: measurement and characterization by electron spin resonance. AB - The concentration of oxygen within cells is important in many physiological and pathological processes, but such oxygen-dependent processes are generally studied as a function of the concentration of extracellular oxygen, due to a lack of suitable methods. Using a newly developed technique based on ESR spectroscopy, we show that respiration stimulation of a cell suspension can result in a significant difference between average intracellular and extracellular concentrations of oxygen. These results indicate that studies of oxygen-dependent phenomena in cells may require measurement of intracellular oxygen concentrations and imply that there are mechanisms in cells that restrict the free diffusion of oxygen. PMID- 2550473 TI - Evidence that alterations in small molecule permeability are involved in the Clostridium perfringens type A enterotoxin-induced inhibition of macromolecular synthesis in Vero cells. AB - The mechanism by which Clostridium perfringens enterotoxin (CPE) simultaneously inhibits RNA, DNA, and protein synthesis is unknown. In the current study the possible involvement of small molecule permeability alterations in CPE-induced inhibition of macromolecular synthesis was examined. Vero cells CPE-treated in minimal essential medium (MEM) completely ceased net precursor incorporation into RNA and protein within 15 minutes of CPE treatment. However, RNA and protein synthesis continued for at least 30 minutes in Vero cells CPE-treated in buffer (ICIB) approximating intracellular concentrations of most ions. Addition of intracellular concentrations of amino acids to ICIB (ICIB-AA) caused a further small but detectable increase in protein synthesis in CPE-treated cells. ICIB did not affect CPE-specific binding levels or rates. Similar small molecule permeability changes (i.e., 86Rb-release) were observed in cells CPE-treated in either ICIB or in Hanks' balanced salt solution. Collectively these findings suggest that CPE-treatment of cells in ICIB-AA ameliorates CPE-induced changes in intracellular concentrations of ions and amino acids and permits the continuation of RNA and protein synthesis. These results are consistent with and support the hypothesis that permeability alterations for small molecules are involved in the CPE-induced inhibition of precursor incorporation into macromolecules in Vero cells. PMID- 2550475 TI - Heparin-derived oligosaccharides: affinity for acidic fibroblast growth factor and effect on its growth-promoting activity for human endothelial cells. AB - The minimal structural requirements for the interaction of heparin with acidic fibroblast growth factor (aFGF) were investigated. Oligosaccharides (tetra- to decasaccharides) obtained by nitrous acid depolymerisation of standard heparin were separated by affinity chromatography on Sepharose-immobilised aFGF. The shortest fragment retained by the affinity column at 0.2 M NaCl and eluted at 1 M NaCl was a "regular" hexasaccharide, a trimer of the most abundant disaccharide sequence in heparin. More complex octa- and decasaccharides were also retained by the column. The oligosaccharides eluted by 1 M NaCl from the affinity column ("high-affinity" oligosaccharides) and those washed from the column at 0.2 M NaCl ("low-affinity" oligosaccharides) were compared for their capacity to protect aFGF from proteolysis and to potentiate its mitogenic activity. At a low ionic strength, all oligosaccharides tested, except the "regular" disaccharide, protected aFGF against trypsin and collagenase digestion. At higher ionic strength (greater than 0.2 M NaCl), only high-affinity oligosaccharides showed a protective effect. The high-affinity oligosaccharides (hexa- to decasaccharides) potentiated the mitogenic activity of aFGF, as measured by [3H]thymidine incorporation into DNA of human fibroblasts. The effect of the oligosaccharides on human endothelial cell proliferation was more complex: inhibition of proliferation was observed in the presence of serum and low concentrations of aFGF (1-5 ng/ml) and potentiation in the presence of higher concentrations of aFGF. The potentiating effect increased as a function of molecular size of the heparin fragments and, for a given size, as a function of the anionic charge of the oligosaccharide. Our results suggest that inhibition of cell proliferation by heparin may result from interference with an autocrine basic FGF-like activity. PMID- 2550476 TI - Both homodimeric isoforms of PDGF (AA and BB) have mitogenic and chemotactic activity and stimulate phosphoinositol turnover. AB - Platelet-derived growth factor (PDGF) occurs as three dimeric isoforms, AA, BB, and AB, which were previously shown to bind to two receptors with different isoform-specificity, the A/B-type (binds all three isoforms) and the B-type (binds only PDGF-BB). Results from competition binding experiments with Swiss 3T3 cells suggest the existence of a third receptor type, which recognizes PDGF-AB and PDGF-BB. Furthermore, Swiss 3T3 cells and human dermal fibroblasts express different relative and absolute levels of these receptor types. In particular, Swiss 3T3 cells express 90,000 PDGF-AA binding sites (A/B-receptors) per cell, whereas human fibroblasts express only 20,000 A/B-receptors per cell. All three PDGF isoforms were tested in either cell type for their effect on DNA synthesis. PDGF-BB and PDGF-AA were also tested in Swiss 3T3 cells for their effect on inositol phospholipid metabolism and chemotaxis. Each isoform promoted all three processes dose-dependently, but there were differences in the maximum cellular responses elicited. These responses reflect the capacity of the cells to bind the individual isoforms. These results demonstrate that the previous distinctions in responsiveness to the different PDGF isoforms are primarily a consequence of the differences in the levels of surface expression of the various isoform-specific receptor types, rather than of the differences in the intrinsic activity of these isoforms. Furthermore, these results suggest that all types of PDGF receptors are capable of responding to their respective ligands by mediating phosphoinositide breakdown, chemotactic responses, and DNA synthesis. Whether they exhibit other functional differences remains to be seen. PMID- 2550478 TI - Protein kinase C activation and down-regulation in relation to phorbol ester induced differentiation of SH-SY5Y human neuroblastoma cells. AB - The role of protein kinase C activation in changes in muscarinic receptor functions and in the appearance of biochemical properties characteristic of neuronal cells was studied in SH-SY5Y human neuroblastoma cells induced to differentiate with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA). A decrease in muscarinic receptor sensitivity with respect to agonist induced Ca2+ mobilization and receptor number parallelled the increase in membrane associated protein kinase C (PK-C) activity. These changes occurred during the first 6 h of culture, and they were associated with rounding-up of cells. A subsequent decrease in particulate PK-C activity was followed by an increase in noradrenaline content, the appearance of an electrically excitable membrane, and an increase in the level of neuron-specific enolase. These changes were accompanied by a pronounced neurite outgrowth. 1-(5-Isoquinolinesulphonyl)-2 methylpiperazine (H-7), an inhibitor of PK-C and cyclic nucleotide-dependent protein kinases, enhanced the morphological differentiation induced by TPA, whereas N-(2-guanidinoethyl)-5-isoquinolinesulphonamide (HA-1004), which primarily inhibits cyclic nucleotide-dependent protein kinases, had no effect on the TPA-induced phenotypic differentiation. H-7 inhibited the decrease in muscarinic receptor sensitivity and receptor number, but had no effect on the appearance of the electrically excitable membrane or on the increase in the neuron-specific enolase level. Both H-7 and HA-1004 inhibited the TPA-induced increase in noradrenaline content. PMID- 2550477 TI - Serum-free culture of normal human melanocytes: growth kinetics and growth factor requirements. AB - Normal human epidermal melanocytes were selectively propagated from mixed (keratinocyte-melanocyte) cultures and primary epidermal cell suspensions in serum-free medium, MCDB 153 containing insulin, bovine pituitary extract (BPE), phorbol-12-myristate-13-acetate (PMA), ethanolamine, phosphoethanolamine, and hydrocortisone. Neonatal foreskin melanocytes (NFMs) replicated more readily than adult melanocytes in culture. Early passage NFMs grown in serum-free medium exhibited a population generation time of 24-48 hours. NFMs assumed a less dendritic appearance and were less pigmented than adult melanocytes. PMA or other protein kinase C-activating phorbol esters significantly enhanced mitogenesis of NFMs; however, cAMP-elevating agents were not required for efficient replication of NFMs. Basic fibroblast growth factor (bFGF) was a potent mitogen for NFMs and replaced the requirement for BPE in the culture medium. NFMs expressed a single class of specific, high-affinity receptors for bFGF, exhibiting a Kd = 3 x 10( 11) M and approximately 76,500 receptors/cell. Neither EGF nor TGF-alpha were mitogenic for NFMs, and TGF-beta reversibly inhibited NFM growth. Rapidly growing, early passage NFMs were shown to have cell cycle times of 19.5, 7.5, and 9 hours for G1, S, and G2/M phases of the cell cycle, respectively. Culture of NFMs to confluence or depletion of growth factors from the culture medium caused reversible, G1 phase-specific, cell cycle growth arrest. Senescence of NFMs was associated with irreversible growth arrest in the G1 phase after 40-45 population doublings in culture. Our data demonstrate that basal medium MCDB 153 can be supplemented with defined factors to cultivate selectively two major constituent cell types of the epidermis, the melanocyte and the keratinocyte. PMID- 2550479 TI - Chemotactic peptide receptor-cytoskeletal interactions and functional correlations in differentiated HL-60 cells and human polymorphonuclear leukocytes. AB - We studied the chemotactic peptide receptor/cytoskeletal interactions in HL-60 cells induced to differentiate with different agents and attempted to correlate these observations with the acquisition of different functional responses. Dibutyryl cyclic AMP-treated cells showed rapid superoxide anion production in response to N-formyl-methionyl-leucyl-phenylalanine (FMLP) and slow, sustained response to phorbol myristate acetate (PMA). Retinoic acid-induced cells showed a slow, sustained response to both FMLP and PMA. Interferon-gamma-treated cells produced no superoxide anion on stimulation with FMLP, whereas tumor necrosis factor (TNF)-treated cells showed a slight response. Chemotactic peptide receptor association was the same in the HL-60 cells treated with different agents, despite marked differences in the superoxide anion generation and actin polymerization responses to FMLP and PMA in these cells. In mature neutrophils chemotactic peptide receptor association with the cytoskeleton was not affected by either pertussis or cholera toxin. However, both toxins inhibited FMLP-induced actin polymerization and superoxide anion generation. This suggested involvement of a G-protein similar to Gt, rather than Gi or Gs. Neither toxin had any effect on PMA-induced superoxide anion generation. These observations indicate that receptor association with the cytoskeleton may not have a significant role in affecting signal recognition and response. Among the several possible roles suggested, clearance of the occupied receptors may be the most important role of the cytoskeletal association. HL-60 cells induced to differentiate with different agents (because of their varied functional responses) might prove very useful in dissecting the molecular mechanisms regulating stimulus-induced activation of neutrophils. PMID- 2550480 TI - Modulation of TGF-beta type 1 receptor: flow cytometric detection with biotinylated TGF-beta. AB - Transforming growth factor beta type 1 (TGF-beta 1) was reacted with NHS-biotin to yield a derivative of TGF-beta 1 which was biotinylated on lysine residues. The biotinylated form of TGF-beta 1 was separated from the unreacted material by reverse phase chromatography. In three separate bioassays, the derivatized peptide was as active as the starting material. The use of FITC-avidin in conjunction with flow cytometry demonstrated that the binding of biotinylated TGF beta 1 to its receptor is saturable, competable, and specific. A 100-fold molar excess of underivatized TGF-beta 1 gave 85% inhibition of binding of the biotinylated peptide to the mink lung cell line CCL-64, while TGF-beta 2 showed no inhibition of binding, nor did insulin, calcitonin, or TGF-alpha. Both CCL-64 cells and human umbilical vein endothelial cells showed a density-dependent down regulation of receptor expression in culture. Several factors were examined that might mediate this effect. The down-regulation was shown not to be due to the secretion of an active form of TGF-beta 1. The extracellular matrix from high density cells did not decrease expression of the receptor. Fibronectin, collagen, and gelatin were also unable to signal changes in receptor expression, even though in other systems such matrix components can regulate the responsiveness of cells to TGF-beta 1. Lastly, staining simultaneously for DNA content and TGF-beta 1 receptor expression showed that there was no correlation between cell cycle and receptor levels. PMID- 2550481 TI - Maintenance of proximal and distal cell functions in SV40-transformed tubular cell lines derived from rabbit kidney cortex. AB - This paper reports the preparation and describes the properties of three renal tubular cell lines derived using SV40 infection of primary cultures of rabbit kidney cortical cells, enriched in proximal cells. RC.SV1 was initially derived from cultures grown in the presence of fetal calf serum exhibiting a low degree of proximal differentiation. The cells were subsequently adapted to grow in serum free hormonally defined medium and display basic properties of proximal tubule cells including well-developed apical microvilli, strong expression of brush border hydrolases, Na+-coupled glucose uptake, and increased cyclic AMP production when exposed to PTH. The other two cell lines were derived from cultures in serum-free hormonally defined medium and propagated in the same medium. They are characterized by some common properties including rare and short microvilli, low expression of apical hydrolases, and low or undetectable Na+ dependent glucose uptake, but differ by their abilities to respond by an increase in cAMP to various hormonal stimuli. RC.SV2 cells are sensitive to calcitonin and to a lesser extent to isoproterenol and PTH, suggesting that they may originate from the thick ascending limb of Henle's loop and the bright portion of the distal tubule. RC.SV3 responds essentially to isoproterenol and arginine vasopressin, suggesting a more distal origin (late distal and initial collecting tubule). Emergence of distal cell lines from cultures exhibiting proximal characteristics may be related to distal cell overgrowth as suggested by analysis of growth kinetics and increased Na+/H+ exchanger activity in RC.SV2 compared with RC.SV1. PMID- 2550482 TI - Retinoic acid modulates dome formation by MDCK cells in defined medium. AB - Retinoic acid dramatically increases the size of domes in confluent MDCK monolayers in a hormonally defined medium (medium K-1). After 4-5 days of retinoic acid treatment, enlarged domes began to appear in confluent MDCK monolayers. After 7 days with 3 x 10(-7) M retinoic acid, the majority of the domes in the monolayers were between 27 and 80 x 10(-3) microns 2 in area, whereas in control medium the majority of the domes were between 0 and 9 x 10(-3) microns 2 in area. The dependence of the retinoic acid effect on prostaglandin E1 (PGE1) was examined. In normal MDCK cells, the effects of retinoic acid on dome size were observed only in medium K-1 supplemented with PGE1. This observation indicated that retinoic acid did not elicit its effects simply by stimulating PGE production. In contrast, in monolayers of PGE1-independent MDCK cells, retinoic acid treatment resulted in an increase in dome frequency even in medium K-1 lacking PGE1. This observation can be explained by the elevated cyclic adenosine monophosphate (cAMP) levels in these PGE1-independent MDCK cells. Dibutyryl cAMP resistant MDCK cells, which normally do not form domes in medium K-1, were also studied. Remarkably, the dibutyryl cAMP-resistant MDCK cells were observed to form domes at a significant frequency when medium K-1 was supplemented with retinoic acid. However in medium K-1 lacking PGE1, an effect of retinoic acid on dome formation by dibutyryl cAMP-resistant MDCK monolayers was not observed. The inability of dibutyryl cAMP-resistant MDCK cells to form domes in medium K-1 has previously been attributed to their decreased cAMP-dependent protein kinase activity. The stimulatory effects of retinoic acid on dome formation may possibly be due to an increase in the activity of a particular cAMP-dependent protein kinase or activation of a separate pathway. PMID- 2550483 TI - Functional receptors for nerve growth factor on Ewing's sarcoma and Wilm's tumor cells. AB - Quantification of changes in levels of c-fos RNA was used as an indicator of the presence of functional responses to nerve growth factor in several human non neuronal cell lines which have previously been shown to express high levels of NGF receptors. Four Ewing's sarcomas, one Wilm's tumor, and one melanoma were examined. Of these cell lines, the Ewing's sarcoma IARC-EW1 showed greatly increased levels (10-20-fold) of c-fos RNA after 1 hour of exposure to NGF. Except for the melanoma line, the other tumor lines exhibited small, but reproducible, elevation of c-fos RNA expression. In IARC-EW1 cells, this induction was analyzed for kinetics, dose-response, and suppression by selective inhibitors of NGF action. The results indicate that these cells bear high affinity receptors for NGF, which utilize signal pathways similar to NGF receptors on PC12 cells. Thus, we report new types of cells with functional responses to NGF and indicate that these may constitute a new model which will usefully complement those presently used for studying the mechanism of action of NGF. PMID- 2550484 TI - Tumor necrosis factor causes amplification of arachidonic acid metabolism in response to interleukin 1, bradykinin, and other agonists. AB - Tumor necrosis factor stimulated prostaglandin E2 synthesis in Swiss 3T3 fibroblasts. Interleukin 1 also stimulated prostaglandin synthesis. Simultaneous addition of tumor necrosis factor and interleukin 1 synergistically stimulated prostaglandin synthesis, even when both growth factors were added at what would be supramaximal concentrations by themselves. Several small peptides and nonpeptides rapidly stimulate prostaglandin synthesis in these cells. Pretreatment with tumor necrosis factor synergistically enhanced prostaglandin synthesis in response to bradykinin, bombesin, thrombin, norepinephrine, and platelet-activating factor. Thus, tumor necrosis factor stimulates prostaglandin synthesis and greatly amplifies prostaglandin synthesis in response to other agonists. This finding may have significance in chronic inflammatory diseases such as rheumatoid arthritis in which several hormones and growth factors may synergistically augment eicosanoid synthesis. PMID- 2550485 TI - Temporary complementation of temperature-sensitive mutants of the cell cycle by transfection with a wild-type or a mutant cDNA of ADP/ATP translocase. AB - A number of cell-cycle-specific temperature-sensitive (ts) mutants have been isolated from animal cells, especially Syrian hamster cells. These ts mutants, like cell cycle ts mutants of yeast, can be complemented by specific genes, some of which have been molecularly cloned. We have isolated a cDNA clone that complements TK-ts13 cells, but only temporarily. This clone, called B1, differs from a previously isolated clone (Sekiguchi et al.: EMBO Journal 7:1683-1687, 1988) that specifically complements ts13 cells. In addition, B1 also complemented temporarily three other ts mutants of the cell cycle, tsAF8, ts694, and ts550C cells. These mutants have different mutations since, in cell fusion experiments, they complement each other. Sequencing of the B1 cDNA clone revealed that it was a mutant of human ADP/ATP translocase in which some human sequences at the 5' end have been replaced by SV40 sequences. The wild-type translocase was less effective but could still increase the survival time of cell cycle ts mutants at the restrictive temperature. Using the polymerase chain reaction, it was possible to demonstrate that the B1 plasmid is expressed in TK-ts13 cells undergoing temporary complementation. PMID- 2550486 TI - Specific desensitization of the epidermal growth factor receptor by pp60v-src. AB - BALB/c 3T3 cells infected with a temperature-sensitive mutant (LA90) of RSV have been used to investigate possible heterologous interactions between the pp60v-src tyrosyl kinase and the epidermal growth factor (EGF) and bradykinin receptors. The LA90 pp60v-src exhibits a very rapid activation t1/2 (less than 5 min) of protein kinase activity on decreasing the temperature from 40 degrees C to 35 degrees C. This change in temperature was also found to induce a very rapid decrease in the affinity for 125I-EGF of receptors on the RSV-LA90-infected cells but not of those on control parental cells. However, no significant changes were detected in the binding of 3H-bradykinin to either cell line. Two separable processes control the desensitization of the EGF receptor by pp60v-src, both of which are independent of protein kinase C. The first is rapid and transient, while the second is sensitive to cycloheximide and persists long after inactivation of pp60v-src. PMID- 2550487 TI - Coupling of antagonistic signalling pathways in modulation of neutrophil function. AB - Modulation of neutrophil activation by catecholamines reflects a fine-tuning by coupling inhibitory and stimulatory receptor pathways. The catecholamine isoproterenol (ISO) binds to beta-adrenergic cell surface receptors and thereby inhibits cell responses such as O2- production stimulated by formyl peptides. However, ISO did not inhibit O2- generation activated by 1 microM ionophore A23187, the protein kinase C activators phorbol ester (PMA, 100 ng/ml) and oleoylacetylglycerol (OAG, 50 microM), and the G-protein activator NaF (40 mM). Furthermore, the overall kinetics of oxidant production in the presence of ISO were unchanged when cells were stimulated with PMA, OAG, A23187, and NaF. These results would imply that neither intracellular calcium, the activation of protein kinase C, nor the activation of G-protein are the primary target of the inhibitory pathway. Accordingly, pertussis toxin did not block PMA or NaF stimulated superoxide generation. In contrast, formyl peptide-dependent GTPase activity is inhibited by ISO in sonicated cell preparations. Since ISO increases the cAMP concentration in the cell, the possibility is raised that a cAMP dependent kinase inhibits signal transduction in part by blocking the interaction of this receptor with its G-protein. PMID- 2550488 TI - Microinjection of inositol 1,2-(cyclic)-4,5-trisphosphate, inositol 1,3,4,5 tetrakisphosphate, and inositol 1,4,5-trisphosphate into intact Xenopus oocytes can induce membrane currents independent of extracellular calcium. AB - Inositol phosphate action in an intact cell has been investigated by intracellular microinjection of eight inositol phosphate derivatives into Xenopus laevis oocytes. These cells have calcium-regulated chloride channels but do not have a calcium-induced calcium release system. Microinjection of inositol 1,3,4,5 tetrakisphosphate (IP4), inositol 1,2-(cyclic)-4,5-trisphosphate (cIP3), inositol 1,4,5-trisphosphate (IP3), or inositol 4,5-bisphosphate [(4,5)IP2], open chloride channels to induce a membrane depolarization. However, inositol 1-phosphate (IP1), inositol 1,3,4,5,6-pentakisphosphate (IP5), inositol 1,4-bisphosphate, or inositol 3,4-bisphosphate are unable to induce this depolarization. The depolarization is mimicked by calcium microinjection, inhibited by EGTA coinjection, and is insensitive to removal of extracellular calcium. By means of the depolarization response, the efficacy of various inositol phosphate derivatives are compared. IP3 and cIP3 induce similar half-maximal, biphasic depolarization responses at an intracellular concentration of approximately 90 nM, whereas IP4 induces a mono- or biphasic depolarization at approximately 3400 nM. At concentrations similar to that required for IP3 and cIP3, (4,5)IP2 induces a long-term (greater than 40 min) depolarization. The efficacy (cIP3 = IP3 = (4,5)IP2 much greater than IP4) and action of the various inositol phosphates in an intact cell and their inability to induce meiotic cell division are discussed. PMID- 2550489 TI - Evidence of an auxin-mediated phosphoinositide turnover and an inositol (1,4,5)trisphosphate effect on isolated membranes of Daucus carota L. AB - Microsomal membranes from carrot suspension cells were phosphorylated in vitro with [gamma-32P]ATP. In the presence of submicromolar concentrations of the natural auxin indoleacetic acid (IAA), a rapid, but transient decrease of the [32P] label could be detected in the phospholipid extracts of the membranes. The phytohormone effect was not the result of an inhibition of the lipid phosphorylation reactions, but was caused by a simultaneous release of water soluble compounds, which, according to their chromatographic properties, were assumed to contain inositol polyphosphates. Although the [32P]-labeled lipids, as well as the inositol polyphosphates, were not identified unequivocally by chemical analysis, these findings point to an auxin-mediated control of a phosphoinositidase C-like reaction similar to the hormone-stimulated phosphoinositide response in animals. Exogenously applied inositol (1,4,5)trisphosphate [(1,4,5)IP3] was found to release 45Ca2+ from preloaded membrane vesicles of carrot cells. Both the detection of the auxin-stimulated phosphoinositide response and the (1,4,5)IP3-mediated Ca2+ release on isolated cell membranes offer new experimental approaches for the identification of the putative auxin receptor and its signal transduction pathway. PMID- 2550490 TI - The metabolism of arachidonic and eicosapentaenoic acids in human neutrophils stimulated by A23187 and FMLP. AB - A23187 stimulates the metabolism of endogenous as well as exogenous arachidonic acid (AA) and eicosapentaenolc acid (EPA) to their corresponding leukotrienes in human neutrophils. In contrast, conflicting results have been obtained concerning the effect of FMLP on the metabolism of these fatty acids. In the present study we compared the effect of A23187 and FMLP on the release and metabolism of these fatty acids in neutrophils. Stimulation of neutrophils with A23187, but not with FMLP, resulted in detectable levels of AA in the presence or absence of BW755C (a dual inhibitor of cyclooxygenase and lipoxygenase). The absolute amount of nonesterified AA in the extracts of neutrophils exposed to the agonist A23187 in the presence of BW755C was 20% higher than that obtained in the absence of BW755C, indicating that only a small fraction of the released AA was converted to lipoxygenase products. Furthermore, significant quantities of AA and EPA metabolites were detected only after treatment of neutrophils with A23187, but not with FMLP. Both A23187 and FMLP stimulated the conversion of exogenous EPA to 5-lipoxygenase products, with A23187 being somewhat more effective. In addition, significant differences were noted on the effect of EPA and DHA on the conversion of AA to its metabolites in A23187-stimulated neutrophils. Our results provide strong evidence that the amounts of eicosanoid precursors mobilized in response to FMLP are extremely small, if any, and this appears to be the likely explanation for the lack of eicosanoid detection by HPLC in FMLP-stimulated neutrophils. PMID- 2550491 TI - Further characterization of four lipocortins from human peripheral blood mononuclear cells. AB - Four calcium and phospholipid binding proteins purified from mononuclear cells were characterized for PKC and EGF phosphorylation, actin binding capacity, and partial tissue distribution. Those named 35K, 32K, and 73K are equivalent, respectively, to lipocortin III, endonexin II and the 67 kDa calelectrin; 36K is a fragment of 73K. After purification, 35K and 73K were phosphorylated by protein kinase C in vitro but 36K nor 32K were not. None were phosphorylated by the epidermal growth factor receptor kinase in vitro; 73K bound F-actin in a calcium dependent manner, whereas 35K, 36K, and 32K did not. Using Western blotting analysis, 32K and 73K were detected in high amounts in human lymphocytes, monocytes, liver, and placenta and in rat adrenal medulla; but 32K was not detected in polymorphonuclear cells, and 36K and 35K were detected in high amounts only, respectively, in human blood lymphocytes and polymorphonuclear cells. Thus, 32K and 73K appear to have a wide tissue distribution, whereas 35K has a much more restricted distribution. PMID- 2550492 TI - Effects of bombesin on human small cell lung cancer cells: evidence for a subset of bombesin non-responsive cell lines. AB - The effects of bombesin on three human small cell lung carcinoma cell (SCLC) lines (NCI-H69, NCI-H128, and NCI-H345) have been examined and compared to the effects of the peptide on the mouse fibroblast cell line Swiss 3T3, and the rat pituitary tumor cell line GH3W5. While all three SCLC lines expressed messenger RNA encoding pro-gastrin releasing peptide (GRP), only the NCI-H345 cells expressed detectable membrane receptors for GRP and responded to nanomolar concentrations of bombesin as shown by 125I-GRP binding, total inositol phosphate accumulation, and increased clonal growth in soft agarose. These data show that some SCLC lines are insensitive to bombesin and do not express detectable membrane receptors for GRP. PMID- 2550493 TI - Excitatory amino acid binding sites in the rat hippocampus after transient forebrain ischemia. AB - The influence of transient forebrain ischemia on the temporal alteration of glutamate receptors in the hippocampal formation was analyzed by means of in vitro quantitative receptor autoradiography. We compared the binding of N-methyl D-aspartate (NMDA) receptors using [3H]3-[+/-)2-carboxypiperazin-4-yl)-propyl-1 phosphonic acid (CPP), noncompetitive NMDA antagonist binding sites using [3H]N (1-(2-thienyl)-cyclohexyl)-3,4-piperidine (TCP), and kainate (KA) receptors. In the CA1 subfield of the hippocampus, the number of NMDA receptors and noncompetitive NMDA antagonist binding sites remained constant during the early stage of recirculation when the CA1 pyramidal cells remained histologically intact. A significant reduction of these receptor densities was observed 7 days following ischemia, when NMDA receptors and noncompetitive NMDA antagonist binding sites lost 64 and 29% of their binding sites in the stratum radiatum of the CA1, respectively. The KA receptor density in the CA1 subfield decreased by 44% 7 days after ischemia. Marked loss of the above-mentioned receptors in the CA1 after selective depletion of the CA1 pyramidal cells indicated that NMDA receptors, noncompetitive NMDA antagonist binding sites, and KA receptors in the CA1 are predominantly localized on the CA1 pyramidal cells. NMDA receptor density in the CA3 gradually decreased during the recirculation period. The stratum moleculare of the dentate gyrus, whose structure was histologically intact after ischemic insult, also showed a reduction of NMDA receptors 7 days following ischemia. [3H]KA receptor density in the stratum lucidum of the CA3 and in the hilus also decreased during recirculation. These PMID- 2550494 TI - [Prognostic factors in hepatocarcinoma]. AB - A critical review of the literature is presented concerning prognostic factors in hepatocarcinoma, notably after surgical resection. Several factors would not seem to play an important prognostic role: age, liver function, alpha fetoprotein level or histological staging of the tumor. Other debatable factors include: sex, since females have a better prognosis, the existence of cirrhosis and the surgical margin of security. On the other hand the situation is clear regarding several other factors such as: size of the tumor, the severity of cirrhosis, the number of nodules, the macroscopic existence of a capsule and the presence or otherwise of portal invasion. PMID- 2550495 TI - Comparison of non-ionic detergents for extraction and ion-exchange high performance liquid chromatography of Sendai virus integral membrane proteins. AB - The integral membrane proteins of Sendai virus haemagglutinin-neuraminidase (HN) and fusion protein (F) were extracted from purified virions with 2% of a non ionic detergent, i.e., polyoxyethylene alkyl ethers varying by 8-14 hydrocarbon units in the alkyl chain and by 4-8 ethylene glycol units in the oxyethylene chain. Triton X-100 and octyl glucoside were included as reference detergents. The hydrophile-lipophile balance (HLB) and the critical micelle concentration (CMC) of the detergents were determined. A decrease in length of the oxyethylate by 8-5 ethylene glycol units and an increase in the alkylate by 8-12 hydrocarbon units resulted in higher yields of extracted proteins. The highest yields were obtained for C12E5 with an HLB of 11.7. Yields of extracted protein could be correlated with the HLB values of the polyoxyethylene alkyl ethers. The structural integrity of HN and F was not affected during extraction by either detergent as measured by their reactivity with monoclonal antibodies directed against native HN and F. Extracts were subjected to anion-exchange high performance liquid chromatography (HPLC) on a Mono Q column in the presence of 0.1% of the detergent used for extraction. Eluate fractions were analysed by sodium dodecyl polyacrylamide gel electrophoresis and recoveries of HN and F protein were determined by size-exclusion HPLC. The immunological activity of HN and F was tested in an enzyme-linked immunosorbent assay. The highest recoveries of HN and F (80%) were obtained with C10E5 in the elution buffer. HN and F were partially purified and the immunological activity was well preserved. PMID- 2550497 TI - High-performance liquid chromatographic assay for nicotinamide-adenine dinucleotide kinase. PMID- 2550496 TI - Comparison of ion-exchange high-performance liquid chromatography columns for purification of Sendai virus integral membrane proteins. AB - The recovery and separation of the integral membrane proteins, the haemagglutinin neuraminidase (HN) and the fusion protein (F), from a Sendai virus detergent extract were compared on three different ion-exchange high-performance liquid chromatography (IE-HPLC) columns: Mono Q, TSK DEAE-NPR and Zorbax BioSeries SAX. The detergent, either 1-O-n-octyl-beta-glucopyranoside (octylglucoside) or decyl polyethylene glycol-300 (decyl PEG-300), used for extraction of HN and F proteins from the virions, was also present in the elution buffers at a concentration of 0.1%. Recovery of HN and F proteins was primarily dependent on the detergent present in the eluent, resulting in yields of HN varying from 18 to 28 and 56 to 67%, when octylglucoside and decyl PEG-300, respectively, were used. The highest yield for HN protein was obtained by separation on either a Mono Q or a TSK DEAE NPR column with decyl PEG-300 as the additive. Yields of F protein were lower, and the highest recovery of 46% was found in the presence of decyl PEG-300 by separation on the Mono Q column. Analysis of the fractions by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and by size-exclusion HPLC indicated that the HN protein eluted in the presence of decyl PEG-300 from the Mono Q and the TSK DEAE-NPR columns was obtained in pure form, while the F protein was slightly contaminated with HN. Analysis of the fractions with monoclonal antibodies directed against conformational epitopes of HN and F proteins indicated that after IE-HPLC the conformation of the proteins is largely retained. PMID- 2550498 TI - In vitro assessment of viral antigen content in inactivated aluminum hydroxide adjuvanted vaccines. AB - Antigen capture enzyme immunoassays (ELISA) were developed to assess the antigenic content of inactivated aluminum hydroxide (AH) adjuvanted porcine parvovirus, pseudorabies, and infectious bovine rhinotracheitis vaccines. Reference preparations of these viruses were constructed as a basis for comparison. Because AH-associated ELISA interference was largely circumvented, the need for isotopic or complex antigen-adjuvant desorption methods was eliminated. A 4-parameter logistic model related optical density to vaccine dilution. High correlation coefficients (r) were routinely achieved with commercial monovalent and polyvalent vaccines, and reference preparations. The procedure quantified antigen in both aqueous and AH-associated phases. The method may be generally applicable as a partial substitute for in vivo vaccine potency testing by allowing in vitro estimation of inactivated viral antigenic content in AH adjuvanted vaccines. PMID- 2550499 TI - Comparison of three protein A-gold immune electron microscopy methods for detecting rotaviruses. AB - Three immune electron microscopic (IEM) methods employing protein A-colloidal gold (PAG) were compared with direct electron microscopy (DEM) and direct IEM (DIEM) techniques for their abilities to detect simian rotavirus (SA11) and human rotavirus. Using PAG IEM the number of rotavirus particles per grid square was 10 to 50 times and 2 to 5 times higher than observed with DEM and DIEM, respectively. A comparison of sensitivities among the three PAG IEM methods showed that the trapping efficiency for SA11 and human rotavirus was significantly enhanced by a serum-in-agar (SIA) method. The SIA method appeared to be a more reliable and sensitive alternative for detection and quantitation of viral particles. PMID- 2550501 TI - Methods for generating reagents to examine idiotype networks within antiviral immune responses. AB - The use of anti-idiotypic antibodies to examine and/or modulate the immune response to various viral antigens has the potential to be of use in many diverse systems. This paper details the method and immunologic parameters used in our laboratory to generate and characterize anti-idiotypic antibodies (anti-Id or Ab 2) with specificity for antibodies directed against viral antigens. These anti-Id reagents have been used in our laboratory for studies involving the immune responses to hepatitis B virus and simian virus 40, which we describe here, as well as herpes simplex virus, and the human immunodeficiency virus. We have utilized these anti-Id reagents to examine the fine specificity of the idiotypes on antiviral antibodies in these systems and have attempted to modulate or induce specific antiviral immune responses. It is anticipated that the methods described herein will be helpful in analyzing the immune response in other viral systems including studies involving viral-receptor interactions. PMID- 2550500 TI - Sensitive detection of nucleic acids and protein of human papillomavirus type 6 in respiratory and genital tract papillomata. AB - We have developed a sensitive method to detect and localize HPV-6 viral DNA, mRNA and protein in biopsy specimens of genital and respiratory tract lesions by using in situ hybridization and immunoperoxidase assays on sections of plastic-embedded tissue. This modified in situ hybridization technique, using ultrathin sections and strand-specific 3H-labelled riboprobes, offers the advantages of superior morphological preservation and detection of viral genomes at low copy number with good resolution. This modified immunocytochemistry provides better sensitivity when compared to previous methods using paraffin-embedded materials. In respiratory tract lesions, immunoperoxidase assay detected only a few capsid antigen positive cells, while in the genital tract lesions, there were more capsid antigen positive cells. Southern transfer analyses and in situ hybridizations demonstrated the presence of more viral nucleic acids in genital tract papillomata than respiratory tract papillomata. Epithelial cells throughout the papillomata were infected by HPV-6 as evidenced by positive hybridization, with more viral DNA present in superficial cells. Our results suggest that genital tract epithelium is more permissive for HPV-6 replication than respiratory tract epithelium. Using stand-specific probes synthesized from subgenomic fragments of the HPV-6 genome in conjunction with nuclease digestions, we were able to demonstrate that HPV-6 transcripts specific to open reading frames (ORFs) E6, E7, E1, L1, and L2 occur in maturing superficial cells. In contrast, transcripts specific to ORFs E1, E2, E4, E5a, and E5b could be detected throughout the whole of the epithelium with more signals noted at the basal cell areas. In addition, the distribution of HPV-6 nucleic acids and protein in a carcinoma in situ of the larynx was analyzed. In comparison to benign respiratory tract papillomata, more viral DNA was found in the malignant lesion, but the pattern and distribution of transcription and capsid antigen was similar. PMID- 2550502 TI - Properties of IgG subclasses to human cytomegalovirus. AB - The IgG subclass of antiviral antibodies to human cytomegalovirus (CMV) is mainly of IgG1 type. Most CMV seropositive sera also have virus-specific IgG3, but of a lower titre as measured by enzyme-linked immunosorbent assay (ELISA). We studied the reactivity pattern of these two IgG subclasses to CMV structural polypeptides in order to define how virus-specific IgG1 and IgG3 contribute to the neutralization of CMV. Neutralization of CMV was performed with CMV IgG1 and IgG3 separated from CMV seropositive human sera on a protein A Sepharose gel. Both IgG1 and IgG3 have a neutralizing capacity. IgG3 had a 10-fold better neutralizing effect than IgG1 when related to the ELISA titre. In order to analyse the specific reactivities, CMV Towne virion polypeptides were separated by electrophoresis and transferred to nitrocellulose. Using mouse monoclonals to human IgG1 and IgG3 in combination with a biotin-streptavidin system, the reactivities of the subclasses were examined. IgG1 and IgG3 appeared to react with the same structural polypeptides. The strongest IgG1 reactivities were obtained with CMV polypeptides of apparent molecular weights of 145, 80, 64, 56, 52 and 27.5 kDa. The CMV IgG3 reactivity was restricted compared to IgG1, the strongest and most frequent reactivities occurring to polypeptides of 145 and 80 kDa. PMID- 2550504 TI - Detection of enteroviruses using cDNA and synthetic oligonucleotide probes. AB - This study compares the detection of enterovirus RNA by cDNA probes prepared from both the 5' and 3' end of the genome of coxsackie A21 and B4 with the use of synthetic oligonucleotides prepared from short but highly conserved sequences in the 5' end non-coding region of the picornavirus genome. The cDNA probes detected enteroviruses with a variable level of sensitivity which presumably depended on the degree of genomic homology with the detecting probes. Generally probes from coxsackievirus A21 detected more enteroviruses than did similar probes from coxsackievirus B4. Probes from the 5' end of the genome of both viruses were more sensitive than 3' end probes. In contrast, synthetic oligonucleotides detected all enteroviruses efficiently suggesting that these probes could be useful as 'universal' probes to detect any enterovirus. This paper discusses the application of these probes in the diagnosis and differentiation of enteroviruses. PMID- 2550503 TI - Polymerase chain reaction and viral culture techniques to detect HSV in small volumes of cerebrospinal fluid; an experimental mouse encephalitis study. AB - A technique is described for the detection of HSV-DNA in very small volumes (5-10 microliters) of cerebrospinal fluid (CSF). The method was evaluated in CSF samples of 4-6-week-old mice inoculated with HSV-1 via the corneal route. The sensitivity of the PCR assay was compared with results of spin-amplified viral culture with immunofluorescent visualization (SAC/IF), routine viral culture (RVC) and radioactive dot-blot hybridization (DBA) in CSF samples obtained from other mice. The results show the PCR to be superior over the other techniques: infectious virus or viral DNA in CSF was demonstrated by PCR, SAC/IF, RVC and DBA in 68, 55, 20 and 2.5%, respectively. These results show the feasibility of the PCR for a rapid, non-invasive diagnosis of human HSV-encephalitis. PMID- 2550505 TI - Absence of nonclassical congenital adrenal hyperplasia in patients with precocious adrenarche. AB - We studied 31 patients (28 girls and 3 boys), ranging in age from 3.2-7.9 yr, with precocious adrenarche defined by the presence of early sexual hair development, no signs of virilization, and bone age within +3 SD of the mean for chronological age. To determine if this symptom complex stemmed from any form of nonclassical (late-onset) congenital adrenal hyperplasia, an ACTH stimulation test was performed on each patient using a standard 0.25-mg dose of Cortrosyn, given as an iv bolus. Twelve pubertal children (7 girls and 5 boys) and 18 prepubertal children (11 girls and 7 boys) served as normal controls. Baseline and stimulated 17-hydroxypregnenolone (17-OHPreg), 17-hydroxyprogesterone, (17 OHP), 11-deoxycortisol, dehydroepiandrosterone, androstenedione, testosterone, and cortisol levels were measured. Using published nomogram standards for serum 17-OHP response to ACTH, no child with precocious adrenarche was diagnosed as having nonclassical 21-hydroxylase deficiency. Eight girls, however, had a stimulated 17-OHP value that exceeded the mean response for pubertal and prepubertal controls by more than +2 SD [range, 295-670 ng/dL (8.94-20.3 nmol/L)]. Stimulated 11-deoxycortisol values [less than 400 ng/dL (11.6 nmol/L)] ruled out any cases of nonclassical 11 beta-hydroxylase deficiency. No patient had nonclassical 3 beta-hydroxysteroid dehydrogenase deficiency, as defined by both the stimulated 17-OHPreg and the 17-OHPreg/17-OHP ratio to be more than +2 SD above the mean for pubertal children [1354 ng/dL (41.0 nmol/L) and 10.4, respectively]. In conclusion, we could not provide any biochemical evidence for nonclassical congenital adrenal hyperplasia in a large group of children with precocious adrenarche. PMID- 2550506 TI - Effects of epidermal growth factor on thyroglobulin and adenosine 3',5' monophosphate production by cultured human thyrocytes. AB - While several workers have identified epidermal growth factor (EGF) receptors on human thyroid membranes, very few reports have described EGF effects on intact human thyroid cells in primary culture, and these were short term studies indicating that EGF effects were primarily inhibitory [reduced iodide uptake and thyroglobulin (Tg), T4, and T3 release]. Paradoxically, in vivo EGF stimulates thyroid growth and increases colloid stores. In this study we examined the effects of EGF on cultured thyroid cells in regard to thymidine incorporation, Tg secretion, and cAMP production during a 12-day period. Addition of EGF (0-30 ng/mL) to medium for 6 or 12 days stimulated thymidine incorporation and enhanced Tg synthesis by thyroid cells. However, the profile of Tg release into medium was biphasic. Tg release was inhibited by EGF (0.1-10 ng/mL) during the first 3 days of culture, but the inhibitory effect disappeared by the sixth day, and EGF stimulated Tg release by day 12 and thereafter. EGF enhanced endogenous cAMP levels in thyroid cells, but did not augment TSH-stimulated increases in cAMP production. Our observations of EGF-stimulated growth and inhibited Tg secretion during short term culture are consistent with the findings of earlier studies with nonhuman thyrocytes. However, the later phase of enhanced cAMP levels with stimulation of Tg secretion indicates that EGF may have trophic effects on thyrocytes previously unrecognized because of the short term nature of the studies. These observations suggest an important role for EGF in maintenance of normal thyroid physiology. PMID- 2550507 TI - Expression of two insulin-like growth factor-binding proteins in a human endometrial cancer cell line: structural, immunological, and genetic characterization. AB - Insulin-like growth factors (IGFs) bind with high affinity to specific proteins in human serum, cerebrospinal fluid (CSF), and amniotic fluid. In serum, IGFs are bound to a complex with an apparent mol wt of 150K in which an acidstable binding protein of 53K, termed BP-53, is found. In amniotic fluid, a different binding protein with an apparent mol wt of 25K, termed hBP-25, has been identified. This binding protein is secreted by endometrial cells and has been shown to block the binding of IGFs to their membrane receptors and inhibit the mitogenic action of the IGFs on human choriocarcinoma cells. It has been proposed that hBP-25 modulates the action of IGFs on endometrial tissue, especially during pregnancy. We have identified two binding proteins produced by an endometrial adenocarcinoma, HEC 1A, neither of which is related structurally or genetically to hBP-25. One is present in two glycosylated forms with apparent mol wt of 37K and 40K, both of which are immunoprecipitated with an antiserum made to BP-53. Both forms are reduced to a core protein of 30K upon digestion with endoglycosidase-F. Furthermore, BP-53-specific RNA was detected in HEC 1A cells. A second binding protein with an apparent mol wt of 32K was also detected and did not increase in size upon treatment with endo glycosidase F. These two binding proteins were partially purified by a combination of wheat germ agglutinin and IGF affinity chromatography, and polyclonal antibodies were generated. The polyclonal antiserum specifically recognizes a 32K protein in human CSF, suggesting that the HEC 1A cells produce the same binding protein that is predominant in CSF. The findings suggest that IGF action in the endometrium may be modulated by more than a single binding protein, and that at least three structurally distinct human IGF binding proteins exist. PMID- 2550508 TI - Estrogen synthetase (aromatase) activity in primary culture of human term placental cells: effects of cell preparation, growth medium, and serum on adenosine 3',5'-monophosphate response. AB - The establishment of human term trophoblast cells in culture is dependent on the method of cell preparation, growth medium used, and presence of serum. Using freshly isolated term placental cells, we investigated 1) the effects of two different methods of removal of nontrophoblast cells and two culture media on trophoblast aromatase specific activity, cellular morphology, and hCG secretion over 72 h; and 2) the sensitivity of these hormonal parameters to cAMP added 24 h after the initiation of culture. Under conditions in which aromatase is responsive to cAMP, we studied the effect of removing serum after 24 h in culture with serum. After trypsin digestion of placental villi, isolated trophoblast cells were either treated with ammonium chloride (A) or purified on Percoll density gradients (P) and then grown in monolayer culture with medium 199 plus 10% fetal bovine serum (M) or Dulbecco's Modified Eagle's Medium (DMEM) plus 20% fetal bovine serum (D). Regardless of the method of treatment or growth medium used, aromatase specific activity increased 10- to 15-fold 24 h after plating, continued to increase from 24-48 h, and then decreased (AM) or remained constant (AD, PM, and PD) from 48-72 h. Addition of cAMP significantly increased activity in DMEM-grown cells (PD or AD) at both 48 and 72 h. Aromatase activity in PM cells grown with cAMP increased at 48 h, then decreased to near-control levels by 72 h; however, in AM cells, no response to cAMP was observed relative to control cells. Secretion of hCG was suppressed for the first 48 h in all cultures, but increased by 72 h (greatest increase in AM cultures). cAMP significantly increased hCG secretion 48 h after its addition under all conditions. We further evaluated the response of the Percoll-purified DMEM-grown cells (PD) to cAMP after serum removal at 24 h. Cells deprived of serum showed significantly higher aromatase specific activity over the entire culture period compared with serum grown cells. Secretion of hCG increased 2- or 3-fold in the presence or absence of serum, respectively, after 72 h in culture. cAMP increased aromatase specific activity by 1.8- and 1.4-fold at 48 h and by 2.5- and 2.4-fold at 72 h in serum containing and serum-free cells, respectively. The secretion of hCG increased 11- to 14-fold at 72 h under both serum-containing and serum-free conditions, respectively. The results show that cAMP can act as an intracellular messenger in the regulation of both aromatase activity and hCG secretion.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2550509 TI - Adrenal steroid, cortisol, adrenocorticotropin, and beta-endorphin responses to human corticotropin-releasing hormone stimulation test in normal children and children with premature pubarche. AB - To determine whether CRH affects adrenal androgen, beta-endorphin (B-E), and ACTH secretion in normal children during sexual maturation, 17-hydroxyprogesterone (17 OHP), androstenedione (D4-A), dehydroepiandrosterone (DHEA), DHEA sulfate (DS), cortisol, B-E, and ACTH were measured after an iv injection of 1 microgram/kg human CRH. Children with premature pubarche were similarly analyzed to establish whether this condition is accompanied by altered hormonal responses to CRH. CRH produced consistent increases in ACTH, B-EP, and cortisol blood levels, which were comparable at all age intervals in all groups. 17-OHP increased after CRH injection, but its response linearly with age. D4-A levels were not influenced, while DHEA and DS levels were only partially influenced by CRH. The stimulated D4 A to 17-OHP ratio increased with sexual maturation, whereas ratios of cortisol to 17-OHP and D4-A to DHEA remained constant. Children with premature pubarche had hormonal responses similar in magnitude to those of prepubertal children of comparable age. In conclusion, an increase in 17,20-desmolase efficiency occurs with postnatal maturation after CRH challenge. Moreover, CRH does not appear to play an important role in premature pubarche. PMID- 2550510 TI - Thyroid-stimulating activity in sera of normal pregnant women. AB - To evaluate the effect of a thyroid stimulator on thyroid function in the sera of normal pregnant women, we measured thyroid-stimulating activity (TSA) using a highly sensitive bioassay based on cAMP accumulation in cultured rat FRTL-5 thyroid cells. Serum was pretreated with 10% polyethylene glycol (PEG), and the supernatant (PEG-pretreated serum) was then used in the following studies. FRTL-5 cells were preincubated in 5H medium and incubated for 2 h with PEG pretreated serum, and cAMP was measured. All 11 patients with untreated hyperthyroid Graves' disease with strongly positive thyroid-stimulating antibody activity had normal TSA, because only 5.6% of their immunoglobulin G was recovered in the PEG pretreated serum. In 32 normal pregnant women, 29 (91%) had positive TSA. Their TSA showed statistically significant positive correlations with serum hCG and free T4 levels, and a negative correlation with serum TSH levels. Moreover, when hCG was absorbed from sera by incubation with the solid phase anti-HCG monoclonal antibody, a significant positive correlation was observed between the rate of decrease in hCG and that in TSA. In conclusion, 1) TSA exists in the sera of normal pregnant women, which reflects hCG itself; and 2) thyroid glands of normal pregnant women may be stimulated by TSA to induce a slight suppression of TSH but not sufficient to induce overt hyperthyroidism. PMID- 2550511 TI - The new aromatase inhibitor CGS-16949A suppresses aldosterone and cortisol production by human adrenal cells in vitro. AB - CGS-16949A is a new orally active nonsteroidal aromatase inhibitor which is more than 100-fold more potent than aminoglutethimide. This compound is an imidazole derivative, and therefore, its possible effect on cytochrome P-450-dependent enzyme activities in the adrenal gland was evaluated. In vitro investigations with dispersed normal and hyperplastic human adrenocortical cells showed that CGS 16949A at 10(-7)-10(-6) M is a potent 11 beta-hydroxylase inhibitor, which inhibits ACTH-stimulated cortisol release to a similar extent as an equimolar concentration of metyrapone (IC50 for both compounds, 10(-7)-5 X 10(-7) M). Etomidate was a more potent 11 beta-hydroxylase inhibitor (IC50, approximately 10(-8) M), while 10(-7)-10(-6) M ketoconazole caused (via 17 alpha-hydroxylase inhibition) a similar inhibition of cortisol release as 10(-7) M CGS-16949A (IC50, 10(-7)-5 X 10(-7) M). The 11 beta-hydroxylase inhibition by CGS-16949A was accompanied by a dose-dependent increase in the release of precursor steroids by the adrenocortical cells in vitro, including deoxycortisol, 17 hydroxyprogesterone, and androstenedione. Aldosterone release was suppressed 50% by 10(-9) M CGS-16949A, while the IC50 for cortisol in the same cells was 10(-7) M. Aldosterone release by the dispersed adenoma cells obtained from a patient with primary aldosteronism was also significantly suppressed by CGS-16949A. We concluded that 1) the new nonsteroidal aromatase inhibitor CGS 16949A is an inhibitor of 11 beta-hydroxylase which is equipotent to metyrapone. At present it is unclear whether the compound at the dose that causes complete aromatase inhibition in vivo also affects stress-induced cortisol release in man. 2) CGS 16949A exerts a very potent inhibitory effect on normal aldosterone release (IC50, 10(-9) M) and on tumorous aldosterone secretion. CGS-16949A might, therefore, be a drug that can be used in the treatment of primary hyperaldosteronism. PMID- 2550512 TI - No correlation between erythrocyte Na,K-ATPase activity and resting metabolic rate in humans. AB - The relationship between erythrocyte Na,K-ATPase activity (ouabain-inhibited 86Rb uptake) and metabolic efficiency (defined as the resting metabolic rate adjusted for body composition) was examined in 31 healthy young subjects (16 men and 15 women, 19-33 yr old). Mean (+/- SEM) Na,K-ATPase activity (expressed as nanomoles of Rb taken up per h/billion erythrocytes) was similar in the men (86 +/- 3) and women (94 +/- 7). After adjusting metabolic rate for body cell mass (total body potassium), men and women had similar metabolic rates, and subjects with the highest percent body fat tended to have the highest metabolic rates. Ouabain sensitive erythrocyte Rb uptake was related to less than 2% of the variability in resting metabolic rate after adjusting metabolic rate for differences in body composition. Erythrocyte Na,K-ATPase activity does not appear to be a useful marker of metabolic efficiency in man. PMID- 2550513 TI - Refractoriness to follicle-stimulating hormone in rat ovarian granulosa cells--a concentration- and time-dependent phenomenon. AB - Granulosa cells from pre-ovulatory ovarian follicles of rats were exposed, in vitro, to one or two pulses of follicle-stimulating hormone (FSH) in a superfusion apparatus. The superfusate was analysed for cyclic adenosine-3',5' monophosphate (cAMP) and steroids. In experiments with two consecutive FSH pulses, the response to the second pulse (100 ng/ml) was inversely related to the concentration of the first FSH pulse (2.5-100 ng/ml). Within certain limits, a lower total amount of cAMP and progesterone was accordingly released in response to the two FSH pulses when the concentration of the first pulse was increased. This refractoriness declined within a few hours as shown in experiments with extended pulse intervals. Two short and separate FSH pulses also evoked a higher combined response than did a single long FSH pulse lasting the whole time period. In these cases the cells thus responded with a lower total cAMP and progesterone release when exposed to a higher amount of FSH. Clinical trials on i.m. versus pulsatile i.v. FSH administration to anovulatory women have shown a similar relationship between FSH dose and effect, though other parameters were measured than those in the present study. PMID- 2550514 TI - Application of numerical systematics to the phenotypic differentiation of legionellae. AB - A total of 163 strains, including 106 strains of Legionella pneumophila, 28 strains of Tatlockia micdadei, and 29 strains of other legionellae (including members of the proposed genus Fluoribacter), were studied. Ten tests which together could distinguish the genera previously proposed were identified. These tests included catalase-peroxidase, gelatinase, hippurate hydrolysis, starch hydrolysis, medium browning, acetoin production, oxidase, medium fluorescence, colony fluorescence, and the bromcresol purple spot test. T. micdadei strains were strongly catalase positive and bromcresol purple spot test positive and produced acetoin but otherwise were usually inert in the other tests. L. pneumophila and Fluoribacter species could usually be distinguished by strength of catalase activity, blue-white colony fluorescence (if present), and differences in frequency of hippurate hydrolysis, starch hydrolysis, yellow-green medium fluorescence, and, to a lesser extent, oxidase activity. With a simple algorithm and computer program, the overall accuracy was 98.8%. PMID- 2550515 TI - Subgroup and serotype distributions of human, bovine, and porcine rotavirus in Thailand. AB - The subgroup and serotype specificities of human, bovine, and porcine group A rotaviruses in stool specimens collected in Thailand were examined by an enzyme linked immunosorbent assay by using subgroup- and serotype-specific monoclonal antibodies. A clear yearly change was observed in the serotype distribution of human rotavirus. Between 1983 and 1984, serotype 4 was the most prevalent, while the highest frequency of serotype 2 was found between 1987 and 1988. All the bovine and porcine rotaviruses examined showed subgroup I specificities and long RNA patterns. It was of note that serotype 3 porcine rotaviruses were found at a high frequency. PMID- 2550516 TI - Diagnosis of herpes simplex virus encephalitis by detection of virus-specific immunoglobulins A and G in serum and cerebrospinal fluid by using an antibody capture enzyme-linked immunosorbent assay. AB - An enzyme-linked immunosorbent assay was evaluated for detection of intrathecal synthesis of immunoglobulin G (IgG) and IgA antibodies to herpes simplex virus (HSV) in patients with HSV encephalitis (HSVE). Since the antibody-capture principle was used and the assay was carried out at the saturation level of the anti-IgG- or anti-IgA-coated solid phase, correction for blood-brain barrier leakage was not needed. A total of 34 pairs of serum and cerebrospinal fluid specimens obtained from 20 patients with HSVE were examined. Intrathecal synthesis of HSV IgG and IgA was detected from day 7 after the onset of illness in patients with HSVE. Specimens from all 19 patients from whom paired serum and cerebrospinal fluid specimens were obtained at more than 10 days after the onset of illness were positive. Intrathecal synthesis of HSV IgG and IgA was not detected in patients with HSVE before day 7 of illness or in any of the 16 control patients with other causes of (meningo)encephalitis. Use of the antibody capture enzyme-linked immunosorbent assay for HSV IgG and IgA allows the rapid diagnosis of HSVE during the second week of illness. PMID- 2550517 TI - Use of low-frequency-cleavage restriction endonucleases for DNA analysis in epidemiological investigations of nosocomial bacterial infections. AB - Epidemiological investigations of bacterial infections are generally based on multiple phenotypic markers that are often difficult to verify. A more general and reliable method is genomic DNA analysis by restriction endonucleases. However, the commonly used endonucleases produce too many fragments for correct separation by agarose electrophoresis. In contrast, simple electrophoretic patterns are obtained after genomic DNA digestion by low-frequency-cleavage restriction endonucleases and pulsed-field gel electrophoresis, making it easier to compare numerous strains from the same species. This technique was used to investigate an Acinetobacter calcoaceticus outbreak in a urologic department and bronchial colonization of artificially ventilated patients by Pseudomonas aeruginosa in an intensive care unit. The method allowed a clear distinction between epidemic and self-contaminating strains in these different epidemiological situations. PMID- 2550518 TI - Prospective study of community-acquired rotavirus infection. AB - We determined titers of group A rotavirus common antibodies and neutralizing antibodies against serotypes 1 to 4 of prototype human rotavirus (HRV) in cord blood and serum specimens obtained from 38 infants at 4-month intervals from birth until 2 years of age. Nineteen of the infants developed one episode of HRV diarrhea each, and they were matched by age and birth weight with the other 19 infants, who did not develop HRV diarrhea during the follow-up period. We estimated the incidence rate of HRV infection for the two groups of infants combined to be a minimum of 1.34 episodes per infant per year, which is 22 times more common than the occurrence of overt disease caused by the virus in this community. The infection occurred constantly throughout the first 2 years of infancy, whereas all but one of the 19 episodes of overt disease occurred before 12 months of age. Seven of these overt episodes were preceded by at least one episode of subclinical infection earlier, and the other seven were probably due to primary HRV infection. The remaining five episodes occurred before 4 months of age, so that we could not ascertain whether they were due to primary infections because of the presence of maternal antibodies. We showed that levels of HRV antibodies in serum specimens obtained before clinical onset of diarrhea varied widely, and, for most infants in the diarrheal group, levels of these antibodies were similar to those in the serum specimens obtained at the same times from the corresponding age- and birth weight-matched control infants. Nevertheless, the age at which overt disease caused by HRV was most prevalent coincided with the time when the maternal antibodies had declined to low levels but the infants had not yet acquired high titers of these antibodies in their sera. PMID- 2550519 TI - Effect of age of shell vial monolayers on detection of cytomegalovirus from urine specimens. AB - The effect of age of MRC-5 cell monolayers in shell vials on the detection of cytomegalovirus (CMV) from urine was evaluated. When the AD169 strain of CMV was used, 8-day-old monolayers had a higher mean count of fluorescent foci than 15 day-old monolayers did (5.78 versus 2.86) (P less than 0.02) and were more frequently positive (21 of 23 shell vials versus 14 of 22 shell vials) (P less than 0.04). Commercial shell vials used for clinical specimens were evaluated in groups of 8- to 11-, 12- to 15-, and 8- to 15-day-old monolayers. When compared with laboratory-prepared shell vials ranging in age from 3 to 9 days, commercial shell vials had a lower number of fluorescent foci in all groups (P less than 0.03, P less than 0.0001, and P less than 0.0001, respectively), the 12- to 15- and 8- to 15-day-old groups were less frequently positive (P less than or equal to 0.02 and P less than 0.02, respectively), and all three groups were more susceptible to the toxic effects of urine (P less than 0.0001, P less than 0.01, and P less than 0.0001, respectively). For all 191 specimens cultured (8- to 15 day-old group), one or both monolayers were destroyed in 60 (31.4%) specimens compared with 9 (4.7%) specimens toxic to laboratory-prepared shell vials (P less than 0.0001). Both the decreased sensitivity of older MRC-5 cells and the increased sensitivity to the toxic effects of urine made commercial shell vial less sensitive than laboratory-prepared shell vials for the detection of CMV. PMID- 2550521 TI - Comparison of three kinds of blood and two incubation atmospheres for cultivation of Bordetella pertussis on charcoal agar. AB - We compared the growth of Bordetella pertussis strains (n = 32) on antibiotic free and cephalexin (40 micrograms/ml)-containing charcoal agar supplemented with 10% defibrinated horse blood, defibrinated sheep blood, or anticoagulant containing human blood. Plates were incubated either in air or in an atmosphere with 5 to 10% CO2. As assessed by mean colony numbers and rapidity of growth, normal air was preferable to CO2 enrichment for incubation. Growth on horse blood agar was more abundant and more rapid than on sheep blood agar, but the difference in general was not statistically significant. Human blood was clearly inferior to both horse and sheep blood. PMID- 2550520 TI - Identification of serotype 9 human rotavirus by enzyme-linked immunosorbent assay with monoclonal antibodies. AB - Hybridomas producing monoclonal antibodies to VP7, a major neutralizing protein of serotype 9 rotavirus (strain W161), were prepared. One monoclonal antibody, W161-6A1, was shown to neutralize only serotype 9 rotavirus strains and reacted specifically with serotype 9 rotaviruses in an enzyme-linked immunosorbent assay. The development of an immunoassay for detection of serotype 9 rotaviruses should facilitate epidemiologic studies. PMID- 2550522 TI - Sonographic relationship between gallbladder wall thickness and the etiology of ascites. AB - Gallbladder wall thickness (GBWT) and serum albumin were determined in 54 patients with ascites. The statistical analysis of the results reveals a significant difference in GBWT between benign and malignant conditions (p less than 0.01). GBWT was significantly thicker in patients with cirrhosis than in those with malignant ascites (p less than 0.01) and other benign conditions. However, no significant differences were found between either cirrhotic cases with and without overlying hepatocarcinoma, or between noncirrhotic patients, malignant or otherwise. On comparing GBWT and simple routine sonography in establishing the etiology of ascites, the diagnostic reliability indices decreased with GBWT. This shows that GBWT determination lacks diagnostic importance in routine practice. No correlation was found between serum albumin and GBWT, which suggests that the increase in GBWT observed in cirrhotic patients is mainly the result of accompanying portal hypertension. PMID- 2550523 TI - Mesodermal mixed tumor of the uterine cervix associated with retroperitoneal lymph node metastasis. PMID- 2550524 TI - An evaluation of 6 dentifrice formulations for supragingival anticalculus and antiplaque activity. AB - A 4 week, double blind clinical trial was conducted to assess the antiplaque/anticalculus activity of test dentifrices containing varying levels of zinc citrate. Subjects were divided into 6 groups, 4 experimental, 1 positive control and a placebo group. All subjects only brushed at home using the placebo control during study weeks 1 and 3. Plaque and calculus were collected at the end of study weeks 2 and 4 on mylar strips worn on lower incisor teeth. Dentifrice efficacy was assessed by comparing group dry and ash weight decrements. While there were no significant differences between the test and control groups, there was a demonstrable trend toward greater inhibition with higher zinc citrate levels, especially among subjects with high levels of plaque and calculus at baseline. PMID- 2550525 TI - Antiretroviral therapy. AB - Research into the biology of human immunodeficiency virus type 1 (HIV-1), the causative agent of the acquired immunodeficiency syndrome, has yielded valuable information about the replicative cycle of the virus. The steps involved are (1) attachment of the virus to a receptor protein on the surface of the target cell, (2) entry of the virus into the cell, (3) replication of the viral genome through a DNA intermediary, (4) entry of proviral DNA into the nucleus of the host cell, (5) integration of proviral DNA into the host cell genome, (6) transcription of proviral DNA to viral genomic and messenger RNA, (7) translation of viral messenger RNA to viral proteins, and (8) assembly of viral components into new virions that are released by budding from the host cell membrane. Each step in the HIV-1 replicative cycle offers a potential target for antiviral chemotherapy. Although many drugs have been developed, none appears singularly effective against all stages of HIV-1 infection. Many obstacles remain in the quest for an effective vaccine against HIV-1. PMID- 2550526 TI - Is the origin of atopy linked to deficient conversion of omega-6-fatty acids to prostaglandin E1? AB - Our hypothesis on the origin of atopy links alterations in omega-6-fatty acid metabolism in atopic persons (i.e., reduced formation of delta-6-desaturase products) to deficient T cell differentiation and function. We suggest that a relative deficiency in dihomo-gamma-linolenic acid-derived prostaglandin E1 is the major etiologic factor for diminished T cell maturation postpartum. Its precursors, gamma-linolenic acid and dihomo-gamma-linolenic acid, are physiologically provided in colostrum and mature breast milk of healthy mothers. Depressed cell-mediated immunity and uncontrolled B-cell response with increased IgE synthesis are explained as prostaglandin E1-dependent defects of T cell differentiation caused by insufficient supply of prostaglandin E1 precursors during early infancy. Thus, in our opinion atopy is a metabolic disorder and the associated immunologic disturbances are epiphenomena. PMID- 2550527 TI - Harderian glands of golden hamsters: morphological and biochemical responses to thyroid hormones. AB - Manipulation of circulating levels of thyroid hormones modifies Harderian gland structure and porphyrin concentrations in male and female golden hamsters. Specifically, thyroxine (T4) and triiodothyronine (T3) induce the morphological conversion of the Harderian glands of females to approximate those of the male. Further, porphyrin concentrations are markedly decreased by this treatment. This effect occurs in ovariectomized animals as well, indicating that the gonads are not involved. Suppression of thyroid function by potassium perchlorate (KClO4) drastically reduces Harderian gland weight in both males and females. However, KClO4 decreases porphyrin levels in the Harderian glands of females and increases it in the male. Concurrently, KClO4 also induces a morphological conversion of the Harderian glands of males to the female type. This effect is evident in photoperiods of either 14:10 (h) or 8:16 (h). PMID- 2550528 TI - MR findings on primitive neuroectodermal tumors. AB - Primitive neuroectodermal tumors (PNETs) are highly cellular primitive CNS neoplasms that resemble microscopically the undifferentiated cells of the germinal matrix of the primitive neural tube. These tumors exhibit a highly malignant behavior with a tendency to disseminate along the CSF pathways. Previous descriptions of the neuroradiological findings in patients with PNET were published prior to the clinical application of magnetic resonance (MR) imaging. We describe our experience with the MR evaluation of four patients with pathologically proven PNET. PMID- 2550529 TI - MR imaging of parotid mass lesions: attempts at histopathologic differentiation. AB - Thirty-four parotid lesions were examined with magnetic resonance imaging. The lesions were studied for a number of factors including size, T1 signal (relative to muscle), T2 signal (relative to CSF), homogeneity, associated adenopathy, associated abnormality of the deep cervical fascia, infiltration of subcutaneous fat, and seventh nerve involvement. Lesions histologically diagnosed as pleomorphic adenoma (benign mixed cell tumor) were predominantly homogeneous as well as smoothly marginated. Those diagnosed as papillary cystadenoma lymphomatosum (Warthin tumor), although equally well marginated, were strikingly heterogeneous, often with areas of hemorrhage. Malignancies and inflammatory masses were both associated with cervical lymphadenopathy; however, inflammatory masses demonstrated fascial thickening of the deep cervical fascia and infiltration of subcutaneous fat ("dirty" fat). PMID- 2550530 TI - Disruption of the blood-brain barrier caused by nonionic contrast medium used for abdominal angiography: CT demonstration. AB - We describe a case of disruption of the blood-brain barrier (BBB) by nonionic, low osmolality contrast medium after abdominal angiography. One plausible explanation of this event is that acute hypertension during angiography may have increased the BBB permeability, allowing the contrast medium to leak across it. PMID- 2550532 TI - Relative adherence of Bacteroides species and strains to Actinomyces viscosus on saliva-coated hydroxyapatite. AB - The study was designed to compare the adherence of several Bacteroides species to A. viscosus. Using 3H, we labeled 24 laboratory strains, including 13 Bacteroides species and 11 fresh clinical isolates of three Bacteroides species. Their adherence to A. viscosus bound to a saliva-coated mineral surface was quantified by liquid scintillation. Adherence relative to a standard strain, B. gingivalis 2561, was compared. Among the lab bacteroides, those of B. gingivalis (eight strains) were the greatest binders (mean, 80.5 +/- 12.4%). Strains of other lab bacteroides bound less well (mean, 33.4 +/- 6.3%). The difference in means was statistically significant (p less than 0.01). The mean for B. gingivalis strains was also significantly greater than that for strains of B. intermedius (51.7 +/- 6.2%). Attachment of B. gingivalis was saturable in experiments in which either input concentration or time was the independent variable, indicating that B. gingivalis cells do not accumulate in this vitro simulation of plaque formation by binding to each other. Subculture did not seem to affect the degree of binding. PMID- 2550531 TI - Adsorbed salivary acidic proline-rich proteins contribute to the adhesion of Streptococcus mutans JBP to apatitic surfaces. AB - Experimental pellicles formed on hydroxyapatite (HA) beads from parotid or submandibular saliva promoted the adhesion of Streptococcus mutans JBP cells to a greater extent than did pellicles prepared from buffer, human plasma, or serum. The nature of the salivary components responsible was studied by the preparation of pellicles from fractions of parotid saliva obtained by chromatography on Trisacryl GF 2000 columns. Two groups of fractions promoted attachment of the organism. Components migrating in the high-molecular-weight mucin fraction were most effective, but a later-eluting fraction also possessed adhesion-promoting activity. Subfractionation of the latter material indicated that the adhesion promoting activity was associated with the acidic proline-rich proteins (PRPs). Pellicles prepared from 10-20-micrograms/mL solutions of pure PRP-1 were effective in promoting attachment of S. mutans JBP cells. PRP-3 was less effective, while human salivary statherin, fibrinogen, fibronectin, type 1 collagen, and the amino-terminal tryptic peptide derived from PRP-1 were ineffective. The quantities of 150-residue and 106-residue PRPs and of statherin, which became incorporated into experimental pellicles prepared from saliva, were estimated with use of radiolabeled protein tracers. The data obtained suggest that these proteins compete for similar binding sites on HA, and that their ratios in saliva would therefore influence the quantity of the larger PRPs that become incorporated into the pellicle. Such competition may contribute to the variability observed in the adhesion-promoting activities of different saliva samples. PMID- 2550533 TI - The effect of silicic acid on calcium phosphate precipitation. AB - So that a possible involvement in the mineralization of dental plaque could be investigated, the effects of silicic acid on calcium phosphate precipitation were assessed in vitro. By measuring the decrease in Ca2+ concentration (by means of ion-selective electrodes), we determined both spontaneous precipitation and seeded crystal growth from solutions that contained 1 mmol/L calcium, 7.5 mmol/L phosphate, 50 mmol/L Hepes pH 7.2, and various amounts of silicic acid. Polymerized silicic acid, but not its monomer, was found both to cause a 60% reduction in the lag period that precedes spontaneous precipitation and to enhance the growth rate of seeded hydroxyapatite crystals. Silica suspensions showed effects similar to those of polysilicic acid. In all cases, the precipitated material was found to be hydroxyapatite. Whereas seeded brushite crystals grew slowly without silicic acid, hydroxyapatite was the only mineral detected after crystal growth in the presence of silicic acid. Apparently, polysilicic acid acted as a substrate for hydroxyapatite nucleation, inducing secondary nuclei on both hydroxyapatite and brushite crystals. The finding that polysilicic acid could overcome part of the inhibitory effect of a phosphoprotein on calcium phosphate precipitation gave additional support for the idea that polysilicic acid and silica may promote the formation of dental calculus. PMID- 2550534 TI - Immunohistochemically detectable duct-like structures in benign and malignant eccrine poromas: CEA and involucrin immunostaining. AB - The duct-like structures present in 7 cases of benign and malignant eccrine poroma were examined by immunohistochemical staining for carcinoembryonic antigen (CEA), involucrin, and S-100 protein. The demonstration of CEA and involucrin was helpful in the recognition of these structures. The overt immunopositivity precedes morphological evidence for duct formation. On the basis of the CEA immunostaining, the duct-like structures were divided into 4 types: 1) mature acrosyringeal structure, 2) cystic luminal structure lined by elongated cells, 3) immature acrosyringeal structure, and 4) vacuole- or dot-like potential lumen in a single cell. Involucrine was observed in the lining cells of 1) and 2). None of the 4 types showed positive reactivity for S-100 protein, suggesting the irrelevance of these structures to the secretory element of sweat gland. The polymorphism of the ductal formation tended to be more remarkably observed in malignant eccrine poromas than in the cases of benign eccrine poroma and poroepithelioma tested. PMID- 2550535 TI - Rapid assessment of liposomal stability in blood by an aqueous nitroxide spin label. AB - An electron spin resonance method using an aqueous nitroxide spin label, 2,2,6, tetramethyl-piperidine-N-oxyl-4-trimethyl-ammonium, for rapid assessment of liposome stability in blood is presented. The retention of the nitroxide in liposomes is measured by its electron spin resonance signal intensity, a procedure which does not require separation of the sample from the blood. Any nitroxide that is released from the liposomes is reduced by external ascorbic acid which is added to the sample. The method permits kinetic studies on the integrity of liposomes in the presence of destabilizing factors such as detergent, blood, or alteration in temperature. PMID- 2550536 TI - A new neomycin phosphotransferase II solid phase assay in combination with polyacrylamide sodium dodecylsulphate gel electrophoresis. AB - A new general method for the determination of neomycin phosphotransferase (NPT) II (EC 2.7.1.95) activity in cell extracts after separation in SDS-polyacrylamide gels is described. The enzymatic activity of NPT II is restored after SDS polyacrylamide gel electrophoresis by incubating the gel for 3 h (20 mM Tris-HCl buffer, pH 7.4). The enzymatic activity is determined by in situ phosphorylation of aminoglycoside antibiotics bound to solid supports and brought into direct contact with the gel surface. A novel, mechanically stable, negatively charged matrix was synthesized for use in this solid phase enzyme assay and compared to phosphocellulose and carboxymethylcellulose paper. This new method allows the easy and exact determination of the molecular weight of any fusion protein with NPT II by assaying the position of the enzymatic activity in the gel and a consecutive immunological reaction following protein transfer onto nitrocellulose membranes. PMID- 2550537 TI - Standardization of the electron spin resonance method for the proteolytic activity assay. PMID- 2550538 TI - Effects of neuropeptide Y on the renal, mesenteric and hindlimb vascular beds of the conscious rabbit. AB - The effects of neuropeptide Y (NPY, 10 micrograms/kg bolus i.v.) on renal, mesenteric and hindlimb blood flow were determined in intact conscious rabbits with chronically implanted Doppler ultrasonic flow transducers. The role of sympathetic neuro-effectors was assessed using inhibition of peripheral alpha adrenoceptors with phentolamine in each group, and in the renal flow group following chemical sympathectomy with 6-hydroxydopamine. In controls, NPY caused markedly non-uniform peak responses. Renal blood flow fell from 2.16 +/- 0.12 kHz to a minimum of 0.26 +/- 0.07 kHz following NPY administration (P less than 0.05). Mesenteric blood flow was reduced from 2.04 +/- 1.17 to 1.54 +/- 0.11 kHz (P less than 0.05). blood flow increased transiently from 2.33 +/- 0.15 to a peak of 3.33 +/- 0.19 kHz (P less than 0.05). Renal vascular resistance rose by 1189 +/- 309% and mesenteric resistance by 54 +/- 9% (P less than 0.05), while hindlimb resistance fell by 24 +/- 3% (P less than 0.05). Pretreatment with phentolamine accentuated the peak pressor response and the reduction in heart rate induced by NPY administration but had little effect on the local haemodynamic changes in each vascular bed. There was no change in the renal vascular response to NPY following sympathectomy. Indeed, the peak NPY-induced reduction in renal blood flow seen in control animals (87 +/- 4%) was unaffected by either alpha-adrenoceptor inhibition (90 +/- 5%) or by sympathectomy (86 +/- 5%). In conscious rabbits with intact cardiovascular reflexes, pharmacological doses of NPY cause profound renal vasoconstriction with smaller changes in mesenteric and hindlimb flow.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2550539 TI - Plasma concentrations of adrenocorticotropin-related peptides in lambs injected with ovine corticotropin releasing factor or vasopressin. AB - Synthetic ovine corticotropin-releasing factor (oCRF) and arginine vasopressin (AVP) were intravenously injected each alone or in combination (each peptide: 1 microgram/kg body weight) in lambs on days 1, 3, 7 and 20 after birth. Plasma samples were collected just before and 10 and 30 min after injection. Plasma concentrations of cortisol and aldosterone were measured. Adrenocorticotropin (ACTH)-related peptides were isolated by Sephadex G50 column chromatography and measured by radioimmunoassay. Three different peaks with an ACTH immunoreactivity were found in lamb plasma: a "big" ACTH molecular form (Mr = 30,000), an "intermediate" (Mr = 8000) and a "little" (Mr = 4500). In 1 and 3 days-old lambs, both CRF and AVP increased preferentially "intermediate" ACTH. In 7 and 20 days old lambs, an increase in "little" ACTH occurred after CRF whereas "intermediate" ACTH rose after AVP. The rise in plasma levels of different molecular forms of ACTH after stimulation by CRF or AVP could suggest that the biological pathway of ACTH synthesis, storage and release may occur in different intracellular pools or rather in different pituitary cells. Intermediate ACTH stimulated adrenal secretion of cortisol as soon as the first day of postnatal life and increased plasma aldosterone concentration in 7 and 20 day-old lambs. At these stages aldosterone level did not change after a rise in "little" ACTH. PMID- 2550540 TI - Leucocyte intracellular pH and Na+/H+ antiport activity in human hypertension. AB - Hypertension is associated with thickening of the wall of resistance vessels, but the cellular or genetic basis of this is unclear. Cell proliferation and intracellular alkalinization via increased Na+/H+ exchange are linked in the response of tissues to growth factors. To define a possible cellular basis for vascular medial thickening in hypertension, we studied leucocyte intracellular pH, buffering power and Na+/H+ antiport activity in 17 hypertensive and 17 age-, sex- and weight-matched normotensive subjects. The cells from hypertensive subjects were significantly more alkaline [median (range): 7.49 (7.26-7.95) versus 7.39 (7.25-7.53); P less than 0.01], and had a lower buffering power [8.95 (3.05-17.98) versus 12.57 (7.44-19.95) mmol/l per pH unit; P less than 0.02] than those from normotensive subjects. Moreover, the activity of the Na+/H+ antiport was higher when cells were acid-loaded to an intracellular pH of 6.7. The presence of a similar increased activity in vascular smooth muscle cells may be associated with increased cellular proliferation resulting in a thickened media or increased vascular smooth muscle contractility. PMID- 2550541 TI - Platelet cytosolic proton and free calcium concentrations in essential hypertension. AB - Alterations in the metabolism of intracellular messengers, such as calcium and cyclic adenosine 5'-phosphate (cAMP), have been reported in essential hypertension. Since intracellular pH (pHi) participates in the control of fundamental cell functions, we looked for changes in platelet cytosolic H+ concentration [( H+]i) in hypertension and investigated whether or not its impaired metabolism is linked to the calcium handling abnormalities. The fluorescent pH indicator BCECF has been used to evaluate intracellular H+ concentration in platelets, unstimulated ex vivo, from normotensive (n = 20) and hypertensive patients (n = 20). Cytosolic [H+] was 20% lower in hypertensive than in normotensive subjects (49.5 +/- 3.4 and 61.8 +/- 2.2 nmol/l cells, respectively, P less than 0.005; mean pHi values were 7.21 and 7.33, respectively). Platelet cytosolic H+ and free Ca2+ concentrations ([Ca2+]i) were determined in parallel in 15 normotensive and 15 hypertensive patients. [Ca2+]i was found to be 19% higher (P less than 0.01), and [H+]i 22% lower (P less than 0.02), in the hypertensive patients compared with the normotensive subjects. Platelet pHi and [Ca2+]i were increased simultaneously in some hypertensive patients. These results are compatible with the hypothesis of an in vivo activation of platelets in hypertension. If a similar alkalinization exists in smooth muscle cells, it may participate in cell proliferation and in an enhanced sensitivity to agonists, two parameters thought to be involved in blood pressure elevation. PMID- 2550542 TI - Platelet cyclic AMP in essential hypertension. AB - Various abnormalities in platelet metabolism, including increased sensitivity to several aggregating agents, have been described in essential hypertension. Platelet response is controlled by Ca2+ and cyclic AMP-dependent mechanisms (stimulatory and inhibitory, respectively) which oppose one another. In the present study, the cyclic AMP contents of unstimulated platelets were measured by radio-immunoassay and observed to be lower in hypertensive than in normotensive subjects, either in the basal state or after prostaglandin E1 (PGE1) stimulation. In the presence of 7-bromo-1,5,dihydro-3,6-dimethylimidazo [2,1-b] quinazolin 2(3H)-one (Ro 15-2041), a specific inhibitor of phosphodiesterase, the increases in cyclic AMP content were similar in platelets from both groups, indicating that this enzyme was not responsible for the alterations in cyclic AMP metabolism observed in hypertension. Low external Ca2+ reduced basal and PGE1-stimulated cyclic AMP content in both normotensive and hypertensive groups but cyclic AMP levels remained lower in hypertensive patients than in normotensive subjects, indicating that Ca2+ influx is not responsible for this altered metabolism of cyclic AMP in hypertension. These data suggest that the reduced platelet cAMP content may participate in the hyperreactivity to various aggregating agents previously reported to accompany essential hypertension. PMID- 2550543 TI - Antibodies to 5'-nucleotidase (CD73), a glycosyl-phosphatidylinositol-anchored protein, cause human peripheral blood T cells to proliferate. AB - Human peripheral blood T cells were stimulated to proliferate when cultured with submitogenic doses of PMA and goat antibodies to 5'-nucleotidase (5'-NT). The degree of proliferation, as measured by [3H]TdR incorporation on day 3, was similar to that achieved by stimulation with PHA. Anti-5'-NT antibodies had no effect on PHA-induced proliferation. Maximal stimulation was achieved with 0.6 to 1.0 ng/ml of PMA and 125 micrograms/ml of IgG isolated from a goat anti-5'-NT antiserum. Both intact IgG and F(ab')2 fragments were stimulatory. IL-2R expression and IL-2 secretion were also induced by anti-5'-NT antibodies and PMA. Anti-5'-NT-induced proliferation was inhibited greater than 95% by a murine anti IL-2 receptor mAb and required less than 0.3% monocytes. Similar results have been obtained with a murine mAb specific for 5'-NT. As expected, anti-5'-NT antibodies and PMA did not induce the proliferation of ecto-5'-NT-T cells isolated by cell sorting. Pretreatment of total T cells with phosphatidylinositol specific phospholipase C removed an average of 89% of the 5'-NT activity from the cell surface and also inhibited by 83% the ability of the cells to proliferate in response to anti-5'-NT antibodies and PMA. Thus, the activation signal provided by anti-5'-NT antibodies is apparently transduced, in large part, by a form of the enzyme that is attached to the membrane via glycosyl-phosphatidylinositol linkage. These data suggest that 5'-NT may play a role in lymphocyte activation as has been proposed for other glycosyl-phosphatidylinositol-anchored lymphocyte surface proteins. PMID- 2550545 TI - Studies on the differing effects of tumor necrosis factor and lymphotoxin on the growth of several human tumor lines. AB - The relative ability of TNF and lymphotoxin (LT) to inhibit the growth of five human tumor cell lines was examined both in the presence and absence of IFN gamma. Two adenocarcinoma lines, HT-29 and SK-CO-1, were 20- and 320-fold more sensitive to the inhibitory effects of TNF and LT in 3- to 4-day proliferation assays. In contrast, the breast carcinoma line BT-20 showed only a one- to twofold difference. The MCF-7 and ME-180 cell lines exhibited intermediate behavior. These results parallel the reported disparate potencies of TNF and LT in their effects on endothelial cells, hematopoietic development and their abilities to sustain a mixed lymphocyte response. Radiolabeled TNF binding studies showed two classes of receptors (Kd 0.04 to 0.15 nM and 0.2 to 1.0 nM) on the highly sensitive SK-CO-1 line. HT-29 cells also appeared to possess some high affinity-binding sites, whereas the BT-20 line completely lacked the high affinity form. Thus the presence of high affinity-binding sites correlated with increased sensitivity to the antiproliferative effects of TNF. Cold TNF competed with the binding of radiolabeled human TNF three- to fivefold better than LT for binding to all three lines. These relatively small differences between the TNF and LT receptor-binding characteristics are insufficient to explain the dramatic disparity in their antiproliferative properties. Likewise, the absolute concentrations of the unlabeled cytokines necessary to block the binding of 125I TNF were 10- to 150-fold higher than the levels necessary to elicit the biologic response. Thus, the receptor binding data conflict with the growth inhibitory effects. This discrepancy is discussed in terms of either separate receptors for TNF and LT or more complex phenomena such as receptor cooperativity possibly resulting from multivalent interactions with the trimeric form of TNF. PMID- 2550544 TI - Coxsackievirus-B3-induced myocarditis: virus receptor antibodies modulate myocarditis. AB - Two variants of coxsackievirus group B, type 3 (CVB3) differ in ability to induce myocarditis in Balb/cCUM mice. Infection with the highly pathogenic variant (CVB3M) stimulates autoimmunity to normal cardiocyte antigens, and tissue injury results primarily from an autoreactive cytolytic T lymphocyte (ACTL). Animals infected with the less pathogenic CVB3o variant do not develop ACTL, although CVB3o replicates to high titers in the heart and polyclonal neutralizing antisera fail to distinguish between the two variant virions. The present study uses two IgM mAb derived by fusing spleen cells from CVB3M-infected mice with NS-1 cells. These mAb investigate important differences between the virus variants that may explain why only selected infections trigger autoimmunity. mAb 8A6 is a virus neutralizing antibody that prevents infection of HeLa cells and cultured cardiocytes by attaching to the virus. mAb 10A1 also interferes with infection but presumably reacts to the virus receptor on the susceptible cells and shows little or no binding to the virions. While 8A6 is equally effective in neutralizing both CVB3o and CVB3M, suggesting that antigenic epitopes on both variants are either identical or highly cross-reactive, 10A1 distinguishes between the variants, suggesting that the pathogenic and less pathogenic viruses use distinct cell surface receptors. Competitive binding studies using radiolabeled CVB3M and either of the unlabeled variants confirm this hypothesis. Both mAb effectively prevent CVB3M-induced cardiac damage in vivo. mAb 10A1 also inhibits autoreactive ACTL lysis of cardiocytes, indicating that the autoimmune effectors may recognize the virus receptor, and that the receptor utilized by a virus may prove important in triggering auto-sensitization. PMID- 2550546 TI - Further studies of the role of transforming growth factor-beta in human B cell function. AB - This study was designed to address three specific questions in human B cells. First, to determine whether transforming growth factor-beta (TGF-beta)2 has similar biologic effects on B cell function as does TGF-beta 1. Second, to test the hypothesis that TGF-beta 1 is an autocrine growth and differentiation inhibitor. Finally, because multiple receptor species for TGF-beta have been identified on other cell types, to determine by chemical cross-linking and competitive binding studies the nature of the TGF-beta 1 R present on normal and transformed B cells. Exogenous TGF-beta 2 was found to be functionally similar to TGF-beta 1 in its inhibition of factor dependent normal B cell proliferation and Ig secretion. When an antibody, specific for the active form of TGF-beta 1, was added in conjunction with IL-2 to previously stimulated B cell cultures, there was a 14.4 +/- 4.2% increase in B cell proliferation, a 22 +/- 6% increase in IgG production, and a 33 +/- 8.6% increase in IgM production when compared to control cultures. Chemical cross-linking of 125I-TGF-beta 1 to normal B cell membranes identified two major cross-linked species of 65 and 90 kDa. A fivefold excess of unlabeled TGF-beta 1 competitively inhibited the detection of both of these bands while a 50-fold excess of unlabeled TGF-beta 2 did not inhibit the 90-kDa band and only partially inhibited (60%) of the 65-kDa band. Chemical cross-linking of 125I-TGF-beta 1 to transformed B cell membranes identified only a single band of 60 kDa. Scatchard plot analysis of 125I-TGF-beta 1 binding to normal B cells that was competitively inhibited with increasing concentrations of unlabeled TGF-beta 1 revealed both high and low affinity binding sites whereas analysis of 125I-TGF beta 1 binding in the presence of increasing concentrations of unlabeled TGF-beta 2 revealed only low affinity sites. These findings demonstrate that TGF-beta 2 is as effective as TGF-beta 1 in inhibiting human B cell function, that small amounts of active TGF-beta 1 are present endogenously in in vitro cultures which partially inhibit B cell function, that two major TGF-beta 1 R cross-linked complexes of 65 and 90 kDa are present on normal B cells, and that transformation of B cells may be accompanied by changes in the TGF-beta 1 R. PMID- 2550547 TI - Transcellular lipoxygenase metabolism between monocytes and platelets. AB - We have examined the effects of co-culture and in vitro co-stimulation on lipoxygenase metabolism in monocytes and platelets. Monocytes were obtained from the peripheral blood of normal volunteers by discontinuous gradient centrifugation and adherence to tissue culture plastic. Platelets were obtained from the platelet-rich plasma of the same donor. When 10(9) platelets and 2.5 x 10(6) monocytes were co-stimulated with 1 microM A23187, these preparations released greater quantities of 12(S)-hydroxy-10-trans-5,8,14-cis-eicosatetraenoic acid, 5(S),12-(S)dihydroxy-6,10-trans-8,14-cis-eicosatetraenoic acid, and leukotriene C4, 5(S)-hydroxy-6(R)-S-glutathionyl-7,9-trans-11,14-cis eicosatetraenoic (LTC4) when compared with monocytes alone. Release of arachidonic acid, 5-HETE, delta 6-trans-LTB4, and delta 6-trans-12-epi-LTB4 from monocytes was decreased in the presence of platelets. A dose-response curve was constructed and revealed that the above changes became evident when the platelet number exceeded 10(7). Dual radiolabeling experiments with 3H- and 14C arachidonic acid revealed that monocytes provided arachidonic acid, 5-HETE, and LTA4 for further metabolism by the platelet. Monocytes did not metabolize platelet intermediates detectably. In addition, as much as 1.2 microM 12(S) hydroxy-10-trans-5,8,14-cis-eicosatetraenoic acid and 12(S)-hydroperoxy-10-trans 5,8,14-cis-eicosatetraenoic acid had no effect on monocyte lipoxygenase metabolism. Platelets were capable of converting LTA4 to LTC4, but conversion of LTA4 to LTB4 was not detected. We conclude that the monocyte and platelet lipoxygenase pathways undergo a transcellular lipoxygenase interaction that differs from the interaction of the neutrophil and platelet lipoxygenase pathways. In this interaction monocytes provide intermediate substrates for further metabolic conversion by platelets in an unidirectional manner. PMID- 2550548 TI - Polymorphonuclear leukocytes cap a derivative of wheat germ agglutinin upon stimulation with formyl peptide and C5a but not leukotriene B4. AB - Previously, we reported that a derivative of wheat germ agglutinin (termed WGA-D) specifically inhibits human polymorphonuclear leukocyte (PMN) chemotaxis to FMLP by blocking reexpression (or recycling) of formyl peptide receptors. WGA-D (? formyl peptide receptor probe) binds to a protein on the PMN membrane that exhibits the same m.w. as the formyl peptide receptor. Since clustering (i.e., capping) of ligand-receptor complexes most likely precedes their internalization, we examined the ability of normal and stimulated PMN to cap fluoresceinated WGA D. We found that, in contrast to capping of fluoresceinated Con A, PMN cap WGA-D in a chemotactic factor-specific fashion. Fluoresceinated WGA-D (5.0 to 20 micrograms/ml) alone did not induce either PMN shape changes (i.e., activation) or capping. Both FMLP (1 to 1000 nM) and human C5a (0.1 to 1.0 nM) induced PMN to polarize and to cap bound WGA-D, in a concentration-dependent fashion. Interestingly, leukotriene B4 (LTB4) (5.0 nM), while inducing the same degree of PMN polarization as FMLP (100 nM) and C5a (0.5 nM), failed to induce PMN to cap bound WGA-D. In contrast, FMLP (100 nM), C5a (0.5 nM), and LTB4 (5.0 nM) induced PMN to cap bound fluoresceinated Con A (10 micrograms/ml) to the same extent. The effect of suboptimal concentrations of FMLP and C5a on capping of WGA-D by PMN was additive. LTB4 did not enhance either FMLP or C5a-induced capping of WGA-D by PMN. Also, FMLP and C5a (but not LTB4) were capable of inducing both desensitization and cross-desensitization of WGA-D capping by PMN. Studies using rhodamine-labeled WGA-D and a fluoresceinated analog of FMLP revealed that both capped to the same place on the PMN membrane. Thus, the data suggest that WGA-D binds to a site on the PMN membrane that is either the FMLP receptor or very closely associated with it. PMID- 2550549 TI - Leukotriene B4 enhances activation, proliferation, and differentiation of human B lymphocytes. AB - Highly purified human tonsillar B lymphocytes at different stages of activation were incubated with leukotriene B4 (LTB4). As a key marker for activation, we used the CD23 Ag. LTB4 enhanced the CD23 expression on resting B cells in synergy with B cell-stimulating factors from 4% to 50%. Maximal effect of LTB4 was observed at 10(-10) M to 10(-12) M. LTB4 also augmented the S and M phase entries as well as Ig secretion in synergy with IL-2 and IL-4. In contrast, 5S,12S dihydroxyeicosatetraenoic acid, an isomer of LTB4, and leukotriene C4 lacked these effects. The results indicate that LTB4 amplifies lymphokine-driven activation, replication, and differentiation of human B lymphocytes. PMID- 2550550 TI - Deficient protein kinase C-dependent Na+/H+ exchanger activity in T cells from bone marrow transplantation recipients. AB - The early Na+/H+ exchanger-mediated alkalinization of intracellular pH (pHi) was analyzed in peripheral blood T cells from 23 bone marrow transplantation (BMT) recipients (17 allogeneic and 6 autologous) and a group of 13 healthy controls, in response to stimulation of protein kinase C (PKC) with a phorbol ester. In parallel we evaluated the proliferative response of peripheral blood T cells to an anti-CD3 mAb in the presence of either IL-2 or PMA. The pHi increase (delta pHi) observed in control samples ranged from 0.14 to 0.23 pH units (X +/- SD = 0.17 +/- 0.03). In 10 allogeneic and four autologous BMT recipients the delta pHi was under the lower limit of the control range (range: 0.01 to 0.09, X +/- SD = 0.05 +/- 0.02), whereas the remaining nine cases responded similarly to control samples (range: 0.14 to 0.24, X +/- SD = 0.17 +/- 0.04). The response of the Na+/H+ antiporter to a PKC-independent osmotic stimulation appeared to be normal, thus indicating that the intrinsic Na+/H+ exchanger activity was unaltered. The anti-CD3 induced proliferative response of the group of samples displaying a suboptimal delta pHi, was significantly lower (p less than 0.01) than that detected in control samples. T cell proliferation in samples from BMT recipients displaying a normal delta pHi was undistinguishable from the control group (p greater than 0.05). Our results provide the first evidence for a defective early metabolic event, closely related to PKC activity, in T cells from BMT recipients displaying a low proliferative response to T cell mitogens. PMID- 2550551 TI - Regulation of lymphocyte tumor necrosis factor receptors by IL-2. AB - Activated lymphocytes are known to express TNF receptors. The precise stimuli involved in induction and regulation of these receptors have not been elucidated. Our findings demonstrate that IL-2, alone and in serum-free conditions, can trigger and regulate TNF receptor expression on normal lymphocytes. Flow cytometric analyses demonstrated that the receptor was rapidly induced on CD4, CD8, and CD16+ cells after IL-2 stimulation. Receptors increased with culture duration, became maximal between days 5 and 9, and were maintained for 18 to 20 days in the presence of IL-2. By using 125I-TNF and FITC-TNF binding, we present evidence that IL-2 concentration determines the magnitude of lymphoid TNF receptor expression--influencing both the percentage of TNF-positive cells within the population and the number of receptors/cell. Collectively, our results are persuasive for consideration of IL-2 as a central mediator in the regulation of lymphocyte TNF receptors. PMID- 2550552 TI - Integration of the transposon-like element ETn upstream of V lambda 2 in the cell line P3X63Ag8. AB - The cell line P3X63Ag8 and hybridomas obtained with it contain two rearranged V lambda 2 EcoRI fragments--6.8 kb and 7.4 kb in size. Southern blot analysis, molecular cloning, and mapping revealed that the rearrangements are related to one another and are not due to normal VJ joining. Sequence analysis indicated that the rearrangement was due to the integration of the transposon like element ETn upstream of V lambda 2. Because a normal embryonic V lambda 2 is still retained in this cell line one of the rearranged fragments must be due to a secondary event after the aneuploidization of the plasmacytoma. RNA dot blot analysis showed that the transposon is strongly expressed not only in embryonic cells but also in plasmacytomas, B lymphomas and T cell lines. PMID- 2550553 TI - Specific binding of a hepatoma nuclear factor to the NF.kappa B/H2TF1 recognition motif found in the C4 promoter, but not in the Slp promoter. AB - The fourth component of C (C4) and sex-limited protein (Slp) genes of FM strain mice show more than 95% nucleotide identity both in the coding and the about 2 kb of 5' flanking region, but are distinct in their regulation. Although C4 is a constitutive gene, Slp is an androgen-dependent gene and shows negligible basal level expression. We have previously shown that this difference in basal expression is determined by a region of about 400 bp immediately 5' of the transcriptional start site of the C4 and Slp genes. In this report, we demonstrate a specific binding site for a HepG2 nuclear extract in this region of the C4 gene by gel retardation and DNase I footprinting experiments. Nucleotide sequence of this binding site is very similar to the recognition sequence of well characterized transcription factors, NF.kappa B and H2TF1. Synthetic oligonucleotides representing the binding sites of the H-2Kb class I and IL-6 genes for these factors compete with the C4 promoter for the complex formation with a HepG2 nuclear factor. Due to the nucleotide deletion and substitution, the corresponding region of the Slp gene lacks this sequence motif as well as binding activity to the HepG2 nuclear factor. PMID- 2550554 TI - Malignant fibrous histiocytoma of larynx. PMID- 2550555 TI - Specificity of anti-H-2 class I antibodies induced by syngeneic immunization with Sendai virus-treated cells is regulated by the mouse MHC and viral antigens. No evidence for MHC-restricted virus-specific antibodies. AB - In a previous study, we searched for Sendai virus (SV)-specific antibodies that were restricted in their binding by self-major histocompatability complex (MHC) antigens. In C57BL/6 (B6; H-2b) mice, most of the sera obtained after i.p. injections with syngeneic SV-coated (SV+) spleen cells contained auto- and alloreactive lymphocytotoxic antibodies directed against H-2 class I molecules, but no viral-specific, MHC-restricted antibodies. Here we report that syngeneic immunization with SV+ cells regularly induced H-2-specific antibodies in various mouse strains. From a total of 12 strains tested, only the B10.S (H-2s) strain appeared to be a low responder. The immune responses are of two types: (i) mice of some strains produce autoreactive antibodies and a broad variety of alloreactive antibodies; and (ii) mice of some strains produce only narrow or widely alloreactive antibodies. Because most of the strains differ only in the H 2 region, the patterns observed are regulated by the MHC. To locate the genes involved in the induction of H-2-specific antibodies more precisely, two B6 mutant strains, bm1 (Kb mutant) and bm13 (Db mutant), were immunized with syngeneic SV+ cells. The results suggest that the H-2Db region plays an important role in the induction and specificity of the lymphocytotoxic H-2 class I-specific antibodies present in sera of H-2b mice after syngeneic immunization with SV+ cells. The role of SV in the induction of H-2-specific antibodies was studied in B6 mice after injections of syngeneic cells coated with liposomes bearing the F and HN proteins of SV. The results suggest that SV surface glycoproteins as well as internal proteins are directly involved in regulating the specificity of anti H-2 antibodies present in sera after syngeneic immunization with SV+ cells. This study does not support the concept that antigen-specific, MHC-restricted antibodies are a part of the B-cell repertoire. PMID- 2550556 TI - Tumorigenic and metastatic ability of SV40-transformed BALB/c cell lines and MHC antigen expression. AB - Tumorigenic and metastatic potential were studied in relation to class I MHC expression in four different SV40-transformed BALB/c cell lines. All the lines studied, tumorigenic or not, expressed both H-2Kd and Dd, so MHC antigens did not seem to be involved in the control of SV40-transformed cells' growth in vivo. Lung metastases were observed in all tumour-bearing mice. Cells cultured after in vivo passage, obtained either from tumour tissue or from individual lung metastases, still expressed similar levels of H-2d antigens, thus suggesting that tumour growth and metastasis do not occur through the selection of variants with altered MHC expression. PMID- 2550557 TI - Susceptibility to acyclovir of herpes simplex virus: emergence of resistance in patients with lymphoid and myeloid neoplasia. AB - We have examined the susceptibility to acyclovir (ACV) of herpes simplex virus isolated from immunocompetent and immunocompromised patients. Susceptibility to ACV was determined in a dye-uptake assay with vero cells grown in 96-well microtitre plates. Resistant strains were found in two out of 35 (5.7%) patients and partially resistant strains in four out of 35 (11.4%) patients immunodeficient as a result of treatment for lymphoid or myeloid malignancy. Strains resistant to ACV were not found in organ transplant recipients or in immunocompetent patients. PMID- 2550558 TI - Disseminated adenovirus infection after liver transplantation and its possible treatment with ganciclovir. PMID- 2550559 TI - Membrane-coating granules are acidic organelles which possess proton pumps. AB - Lysosomes are by definition organelles that maintain an internal acidic pH and contain hydrolytic enzymes. Membrane-coating granules contain a battery of hydrolytic enzymes, in addition to their lamellar discs, and are therefore commonly assumed to be lamellate lysosomes. Although there are data confirming the existence of enzymes in membrane-coating granules, there is no direct evidence to suggest that their internal pH is acidic. As part of a wider program on their role in desquamation, our aim was to determine whether membrane-coating granules are indeed acidic and possess proton pumps. Chloroquine and monensin were selected as the pH markers because both induce swelling of acidic organelles. In four repeat experiments dermatome slices of pig ear skin (2 mm2 x 0.5 mm) were incubated as organ cultures either alone (control) or with 1 mM chloroquine or 25 microM monensin. Ultrastructural observations revealed no swelling in control specimens. In contrast, the inclusion of chloroquine or monensin caused swelling of specific organelles including membrane-coating granules, lysosomes, and trans elements of Golgi stacks, but not mitochondria, rough endoplasmic reticulum, or nuclear envelopes. Swelling of membrane-coating granules and the other organelles was prevented by pretreatment with N,N' dicyclohexylcarbodiimide, a known inhibitor of lysosomal H+ ATPase activity. These findings suggest that membrane-coating granules actively maintain an acidic interior with the aid of proton pumps. Furthermore, membrane-coating granules are heterogeneous because swelling of the whole population did not commence simultaneously. However, it remains to be determined whether this heterogeneity reflects variations in membrane-coating granule pH, leakiness of their membranes to cations, or the number or activity of their proton pumps. PMID- 2550560 TI - Alpha-melanocyte stimulating hormone modulates contact hypersensitivity responsiveness in C57/BL6 mice. AB - The neuropeptide alpha-melanocyte stimulating hormone (alpha-MSH) can act as an antagonist to interleukin 1 (IL-1) bioactivities such as inhibition of fever production, thymocyte proliferation, and inhibition of release of acute phase inflammatory molecules from the liver. In this report we have found that epicutaneous application of alpha-MSH suppresses both the sensitization and elicitation limbs of the cutaneous immune response (CIR) to potent contact sensitizers like dinitrofluorobenzene (DNFB) or oxazalone (OX) in mice. Further, the loss of contact hypersensitivity due to applications of alpha-MSH could be reconstituted by either intradermal or intravenous injections of epidermal thymocyte activating factor (ETAF)/interleukin-1. Topical application of alpha MSH did not cause an alteration in Ia+ dendritic cells (i.e., Langerhans cells) but did produce a significant reduction in the expression of Thy1.2 marker on the Thyl+ dendritic epidermal cells (Thy1+DEC). It has no effects on the phenotypic expression of asialo GM-1 on these same cells. These observations suggest that alpha-MSH, a peptide classically isolated from the pituitary but found in many other tissues and cells of the body, may represent an additional biologic modifier than can modulate suppression of the contact hypersensitivity responses to various haptens. However, the mechanisms by which alpha-MSH or potentially other peptides found in the skin produce these suppressive effects have not been elucidated. PMID- 2550561 TI - The exocrine pancreas in patients with hyperinsulinemic hypoglycemia. A morphometrical and ultrastructural study. AB - The exocrine pancreas has been studied morphometrically and ultrastructurally in 12 patients with insulin-producing tumors in comparison with control patients free from gastrointestinal or pancreatic diseases and with normal glucose homeostasis. Light microscopical examination and morphometrical data revealed that in the insulinoma-bearing patients acinar cells undergo a marked and significant decrease in zymogen granule content, together with a slight reduction in the mean cell area. Such changes were accompanied by ultrastructural features indicating increased production and exocytosis of zymogen granules. The findings concerning centroacinar/ductular cells are consistent with an increase in their number and mean cell area, and an enhancement of their functional activity. In synthesis, both the enzyme- and bicarbonate-secreting cells appear to be greatly stimulated in conditions of chronic hyperinsulinemic hypoglycemia owing to the presence of insulinomas. PMID- 2550563 TI - Fatal clostridial pancreatitis following ERCP and percutaneous needle biopsy. AB - A patient with a large necrotic pancreatic carcinoma underwent ERCP and percutaneous biopsy of the cancer and of a suspected hepatic metastasis. On the second day, she developed a massive hemolysis with a 50% drop of hemoglobin level and a rise of serum bilirubin level from 1.3 to 37.5 mg%. Gas was noted on followup CT scan of the pancreas. Clostridium perfringens was found by needle aspiration of the pancreas and on multiple blood cultures. Death resulted from sepsis on the fourth day. Since ERCP and percutaneous needle biopsy of the pancreas are being done with increasing frequency, and since this complication has not been previously described, this case report is presented. PMID- 2550562 TI - Presence of calbindin-D 28K in endocrine pancreatic tumoral cells of the RINm5F line. AB - Calbindin-D 28K expression in insulin-producing tumoral cells of the RINm5F line was assessed by Western-blot and high pressure liquid chromatography. Western blot analysis demonstrated the presence in RINm5F cell homogenates of a protein recognized by a specific polyclonal antibody against chick calbindin. Proteins with apparent molecular weights (mol wt) of 44, 47, 56, and 85 kD were also recognized by the antiserum in RINm5F cell extract, but not in normal rat islet extract. HPLC heat-resistant protein extract from RINm5F cell homogenates revealed three calbindin positive peaks: a major peak with a retention time (20.5 min) identical to that found in a rat cerebellar extract and two minor peaks with shorter retention times. The calbindin content of RINm5F cells was apparently unaffected after 9 d culture in a medium supplemented with 10% calf serum pretreated with dextran-charcoal to remove 1,25-dihydroxyvitamin D3. PMID- 2550564 TI - [An outbreak of enterocolitis due to Clostridium perfringens in a hospital for the severely disabled]. AB - We had an outbreak of 14 cases of enterocolitis due to Clostridium perfringens (Cl. perfringens) in a hospital for the severe multiply-disabled, where the 100 disabled were admitted, in summer in 1985. The signs and symptoms shown by this enterocolitis were primarily diarrhea without fever and loss of appetite. The feces of 10 cases were examined bacteriologically. The test showed 10(3) to 10(6) cells of Cl. perfringens per one gram of their feces and all the strains isolated were untypable by the classification of Hobbs. Nine out of 10 cases were randomly selected and all of the 9 cases were proved to have enterotoxin producing strains. All the strains were highly sensitive to many kinds of antibiotics except kanamycin and gentamicin. Eleven out of the 14 cases were admitted in the same ward and the 7 out of the 11 cases were in the same room of this ward. Considering the spreading route of this infection, it is unlikely that this outbreak occurred due to food supplied from kitchen in this hospital, because all of the disabled, admitted in this hospital, had little chance by which some of the disabled only in a specific ward or room were supplied with bacteriologically contaminated meals from the point of view of cooking and supplying system of this hospital. Adding to this fact, if this outbreak was due to food-born infection, the symptoms of most patients should occur within 1-2 days, because the incubation period of this disease is within a day, however, the patients increased day by day for more than a week.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550566 TI - [Clinical analysis of 17 cases of Japanese encephalitis experienced in the last ten years]. AB - We studied on the prognosis-related factors on 17 cases of Japanese encephalitis experienced during the last ten years and compared the clinical features with those in previous reports, especially when the disease was prevalent. The patients, ranging from 33 to 91 years old, consisted of 7 men and 10 women. All of them showed an encephalitis type. 47.1% of the patients were completely cured, 35.3% cured with sequelae and 17.6% died. The mortality rate was decreased compared to that of previous reports. All of the death cases were women. There was no relationship between the mortality rate and the patients' age. As previously reported, the more severe the disturbance of consciousness, the higher the mortality was. However the rate of death was lower than that of previous reports. We conclude from this study that clinical features of the disease have changed when compared with that of the older days. PMID- 2550565 TI - Enhancing effect of Salmonella enteritidis SPA on nonspecific resistance. AB - The peritoneal macrophages from mice treated with soluble protective antigen (SPA) or lipopolysaccharide (LPS) showed an increase in chemotactic activity, but muramyl dipeptide (MDP)-induced macrophages did not show any enhancing effect. Conversely, the chemotactic activity of mice peritoneal neutrophils was enhanced by SPA, LPS or MDP treatment. The superoxide anion (O2-)-generating activity of SPA-induced macrophages was higher than LPS or MDP. And even 30 days after SPA treatment, a significant increase of O2(-)-generating activity was evident as compared to the control. SPA-induced macrophages showed a higher degree of intracellular killing of Listeria monocytogenes, in vitro, as compared to macrophages obtained from normal mice. In the studies of in vivo protection, the number of bacteria in the liver after challenging with L. monocytogenes was smaller in SPA-treated mice than in untreated mice. Also, SPA-treated mice showed an increased resistance to L. monocytogenes infection. PMID- 2550567 TI - [An assessment of therapeutic effects by transcatheter arterial embolization (TAE) with degradable starch microspheres (DSM) for hepatic malignant tumors]. AB - Transcatheter arterial embolization (TAE) by using degradable starch microspheres (DSM) as embolic material, 40 microns in diameter and degraded by serum amylase within one hour, was carried out for thirteen cases with hepatic malignancies including eleven cases with hepatocellular carcinoma (HCC), one case with cholangiocarcinoma and one case with metastatic liver cancer. DSM were mixed with anticancer agents and administered through the catheter, which was introduced by Seldinger's method, via the hepatic artery immediately after hepatic angiography in ten cases and through subcutaneously implanted drug delivery system (Port-A Cath) in three cases. The dose of DSM was 900 mg/body and adriamycin 30-40 mg/m2 or 12-14 mg/m2 were used. The former was administered through the catheter immediately after angiography and the latter through Port-A-Cath for HCC and cholangiocarcinoma. A same dose of DSM and mitomycin C 15-16 mg/m2 was administered for metastatic liver cancer through the catheter immediately after angiography. The administration was repeated weekly in three cases in which Port A-Cath was implanted, and at five weeks' interval through the catheter immediately after hepatic angiography in the other ten cases. Therapeutic effects were assessed and pharmacokinetics of adriamycin were studied. Results were as follows; 1) Partial response (PR) was obtained in five cases out of eleven cases with HCC (45.5%) and there were three cases with minor response (MR) in the other six cases. Totally, decrease of tumor size was demonstrated in eight cases out of eleven cases (72.7%). 2) In nine cases in which AFP was positive, the titer of AFP was decreased in seven cases (77.8%). 3) No change (NC) was obtained in cholangiocarcinoma.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550568 TI - [Antitumor effect and indication of chemoembolization using degradable starch microspheres and regional hyperthermia in patients with hepatocellular carcinoma]. AB - Antitumor effects and indications of chemoembolization using degradable starch microspheres (DSM) and regional hyperthermia were investigated in patients with unresectable hepatocellular carcinoma. Chemoembolization using DSM was performed in 40 cases and fourteen cases were treated with the combination of hyperthermia. Tumor regression rate over 50% was 48% (chemoembolization alone: 42%, combined hyperthermia: 57%). Though one and two year survival rates were 62% and 54% respectively in the treatment with chemoembolization alone, one year survival rate was 91% in the treatment with combined hyperthermia. From the viewpoint of antitumor effect, it was thought that effectual indications of chemoembolization using DSM alone were the cases whose tumor size were below 7 cm and portal invasion were negative. These indications however were spread by the combination of hyperthermia. The DSM, temporary embolus, is suitable for combination of hyperthermia because chemoembolization using DSM can be performed many times. The cases with clinical stage III and obstruction of the first branch of portal veins did not become severe hepatic failure because DSM was temporary embolus. Therefore, from the viewpoint of safety, chemoembolization using DSM has almost the same indications as intra-arterial infusion therapy. These results suggested that chemoembolization using DSM will be a beneficial methods as combination therapy for unresectable hepatocellular carcinoma. PMID- 2550569 TI - Intracranial metastasis of hepatic carcinomas: a study of 9 cases within 28 years. AB - A study was made on nine hepatic carcinomas with intracranial metastases. These cases brought a total number of reported intracranial metametastatic hepatic carcinomas to 32 instances. The secondary intracranial hepatic carcinomas formed a proportion of 1.3 to 2.9 per cent among intracranial metastatic tumors. Nine patients in this series were young with an average age of 37 years. There were five hepatocellular carcinomas and four cholangiocarcinomas. An intracranial secondary hepatocellular carcinoma in a 37-year-old man produced massive intracerebral hemorrhage. Only four cases including the current one have been described as massive intracerebral hemorrhage because of secondary hepatocellular carcinoma. Our case, moreover, had a high level of hematocrits representing a paraneoplastic syndrome. PMID- 2550570 TI - Mutational analysis of the virion-sense genes of maize streak virus. AB - Insertion and deletion mutagenesis of the two virion-sense genes, V1 and V2, of maize streak virus (MSV) prevents symptomatic infections following Agrobacterium mediated 'agroinoculation' of maize seedlings. These genes code for an Mr 10900 protein and for coat protein, respectively. Mutants containing insertions or deletions in the coat protein gene, V2, were able to replicate to low levels, producing dsDNA although virion ssDNA was not detected and symptoms were not observed. Hence, unlike the bipartite geminiviruses, MSV requires coat protein to produce symptomatic systemic infection. Mutations in gene V1 which considerably shortened the Mr 10900 protein (V1 gene) resulted either in low levels of replication, in which all the DNA forms associated with a wild-type infection were produced, or in no infection, in which case coat protein production may also have been affected. A V1 mutant generated in vivo with 11 of the 14 N-terminal amino acids altered, was viable and produced symptoms typical of a wild-type infection. Infectivity, assessed by replication and symptom expression, was restored by co-inoculating constructs containing single mutations in different open reading frames, thus rescue can occur by trans-complementation of gene products. The experiments showed that the mutations did not affect the nucleotide sequence requirements for replication and that in all cases intermolecular recombination eventually resulted in dominant wild-type virus. PMID- 2550571 TI - Detection of herpes simplex virus type 1 gene expression in latently and productively infected mouse ganglia using the polymerase chain reaction. AB - The polymerase chain reaction (PCR) was employed to detect herpes simplex virus (HSV) sequences in the DNA, and HSV gene expression in total cell RNA, extracted from cervical and trigeminal ganglia of mice during productive and latent infection with HSV-1, strain SC16. Such gene expression was detected in 1 microgram or less of RNA, the quantity anticipated to be present in one or two cervical ganglia. Within the limits of the primers available, gene expression during latency appeared to be restricted to the latency-associated transcript (LAT). The 195 base portion of the LAT amplified by the PCR was sequenced and found to contain several base changes and deletions with respect to published sequences for different HSV strains. These mutations, within the putative open reading frame 2 of the LAT, formed stop or terminator signals, which suggests that the LAT does not act to establish or maintain latency through translation to a protein. The primers for the LAT also amplified a 300 bp fragment from any murine and some other mammalian RNAs. Apart from the oligonucleotide primers, this fragment did not show any homology with HSV. PMID- 2550572 TI - Clinical and serological outcome of genital herpes simplex virus (HSV) type 2 inoculation following oral HSV type 1 infection in guinea-pigs. AB - The clinical and serological outcome of genital herpes simplex virus type 2 (HSV 2) inoculation in animals previously orally infected with HSV type 1 was evaluated. A prior HSV-1 oral infection modified the genital HSV-2 infection so that only four of 18 (22%) animals were initially symptomatic although all but one animal shed HSV-2 from the cervicovaginal area for at least 5 days following inoculation. Three of four animals with symptomatic initial disease also developed recurrences, as did an additional six animals that did not manifest acute genital disease. Anti-glycoprotein gG-1 antibody was found in 17 of 18 animals with only an HSV-1 infection and anti-gG-2 antibody in all of nine animals with only an HSV-2 infection. Anti-gG-2 antibody was detected in eight of 17 animals with a prior HSV-1 infection following HSV-2 inoculation and one had an indeterminate response. Eight of these nine animals developed recurrent genital disease compared to one of eight that did not respond to gG-2 (P less than 0.006). Thus a prior oral HSV-1 infection modified both the initial presentation of HSV-2 infection and the HSV type-specific serological response. PMID- 2550573 TI - Isolation and preliminary characterization of temperature-sensitive mutants of mouse cytomegalovirus of differing virulence for 1-week-old mice. AB - To study the pathogenicity of murine cytomegalovirus (MCMV) and to identify virulence determinants, we have isolated and phenotypically characterized a set of temperature-sensitive mutants. One mutant, PP269/38, was avirulent for 1-week old BALB/c mice and restricted in its plaque formation and replication at 39 degrees C. Mutants PP242/68 and PP268/38 were 100-fold less virulent than salivary gland-grown virus (SGV), even after two passages in the salivary glands of 1-week-old mice. The former mutant was unable to replicate or form plaques at 39 degrees C whereas the latter replicated poorly at 39 degrees C but not at all at 40 degrees C although it was able to form plaques at 40 degrees C with a reduced plaque size. PP31/15 exhibited a 40-fold reduction in virulence compared to SGV after two passages in vivo and was unable to form plaques or to replicate at 40 degrees C; at 39 degrees C it was able to, with reduced efficiency. The remaining two mutants, PP99/3 and PP392/31, were 10-fold less virulent than SGV and were restricted at 40 degrees C. The six mutants have been classified into at least four complementation groups. These mutants may be useful for studying various aspects of MCMV pathogenicity. PMID- 2550574 TI - Human cytomegalovirus RNAs immunoprecipitated by multiple systemic lupus erythematosus antisera. AB - The association of human cytomegalovirus (HCMV) RNAs with ribonucleoprotein particles that react with antibodies from patients with systemic lupus erythematosus was tested by immunoprecipitation with multiple patients' sera. A major late 2.8 kb RNA and several minor RNAs encoded by the HCMV long repeat region were immunoprecipitated from HCMV-infected cells by La, Ro and, much less abundantly, Sm autoimmune antisera. The exact location of these RNAs was determined by high resolution R-loop mapping and found to be between 0.8093 and 0.8189 map units. The 2.8 kb RNA is polyadenylated and associated with polysomes but does not appear to be spliced. Immunoprecipitation was not seen using normal or other autoimmune antisera. In addition, immunoprecipitation was specific to these RNAs in that other abundant HCMV RNAs were not immunoprecipitated. It was also found that the addition of increasing amounts of purified La antigen to infected cell lysates inhibited immunoprecipitation of the 2.8 kb RNA by La antiserum. The data suggest that specific HCMV RNAs may interact with cellular ribonucleoproteins known to be involved in post-transcriptional regulation of gene expression. PMID- 2550575 TI - Acid stability of hepatitis A virus. AB - The acid stability of unpurified and highly purified hepatitis A virus (HAV) was tested and compared with that of poliovirus type 1, coxsackievirus types A9 and B1 and echovirus type 9. Only HAV had a high residual infectivity after 2 h of exposure to pH 1 at room temperature, remaining infectious for up to 5 h. At 38 degrees C, pH 1, HAV remained infectious for 90 min. Highly purified HAV was found to be infectious for 8 h at pH 1 and room temperature. This indicates that the increased stability is not due to protection by cellular material attached to the virus, but is a virus-specific marker. Under the same conditions, at pH 1 and room temperature, unpurified and highly purified HAV antigens were traceable for 5 and 4 h respectively. PMID- 2550576 TI - Propagation of hepatitis A virus in hybrid cell lines derived from marmoset liver and Vero cells. AB - To establish monkey liver cell lines with a high susceptibility to hepatitis A virus (HAV), marmoset (Saguinus labiatus) liver cells were fused with Vero cells deficient in hypoxanthine-guanine phosphoribosyltransferase and the resulting hybrid cells were selected in HAT medium. Of four hybrid cell lines obtained (S. 1a/Ve-1 to -4), three (S. 1a/Ve-1, -3 and -4) were equally susceptible to HAV infection. When inoculated with a virus isolated from marmoset liver tissue (10% liver tissue extract) or a faecal virus (10% stool extract) from a human hepatitis A patient, all susceptible cell lines showed a significant elevation of viral antigen activity as seen in radioimmunoassay and/or immunofluorescent antibody assays, at 4 to 6 weeks post-infection (p.i.) with the liver-derived inoculum and at 6 to 8 weeks p.i. with the stool-derived inoculum. In S. 1a/Ve-1 cells, a representative of the susceptible hybrid cell lines, full adaptation of HAV (liver tissue virus concentrate) to cell culture was attained after four serial passages. Thereafter, the virus grew to a plateau titre of 10(8.5) TCID50/ml at 7 days p.i. in a growth experiment. The infected cells showed no cytopathic effects but eventually a persistent infection was established when a saturated level of virus growth was reached. PMID- 2550577 TI - The polypeptide of Mr 14,000 of porcine transmissible gastroenteritis virus: gene assignment and intracellular location. AB - Synthetic oligopeptides, corresponding to an amino acid sequence encoded by a potential Mr 9000 product's open reading frame (ORF-4) at the 3' terminus of the transmissible gastroenteritis virus genome, were used to generate rabbit antiserum. These antibodies produced immune complexes with an Mr 14,000 (14K) polypeptide in infected cells. The 14K product was shown by immune fluorescence to become associated with the cell nucleus, correlating with the onset of nuclear vacuolation, and suggesting a role in pathogenesis for the ORF-4 gene. PMID- 2550578 TI - A herpes simplex virus type 1 mutant containing a deletion within immediate early gene 1 is latency-competent in mice. AB - We have investigated the behaviour in mice of the herpes simplex virus type 1 (HSV-1) mutant dl1403, which contains a deletion within the gene encoding the immediate early polypeptide Vmw110. The deletion was responsible for a reduction in virulence assayed by both the intracranial and footpad routes of inoculation. Following injection into the footpad, dl1403 was able to reach the spinal cord and establish a latent infection in sensory ganglia from which virus spontaneously reactivated upon explanation. The Vmw110 polypeptide is therefore dispensable for the establishment and maintenance of latency and for reactivation from the latent state. PMID- 2550579 TI - Antigen-presenting capacity of epidermal cells infected with vaccinia virus recombinants containing the herpes simplex virus glycoprotein D, and protective immunity. AB - We studied the association of herpes simplex type 1 (HSV-1) glycoprotein D (gD-1) expression in epidermal cells (EC) with virus-specific immunity and protection of mice from fatal HSV-2 challenge. Vaccinia virus recombinants containing gD-1 under the control of an early (VP176) or late (VP254) vaccinia virus promoter were used. Mature gD-1 protein was expressed in VP176-infected EC and they had accessory cell function for HSV-2-induced T cell proliferation of immune lymph node cells (LNC). It was not expressed in VP254-infected EC and they did not act as accessory cells. LNC from VP176- but not VP254-immunized mice proliferated in response to HSV antigen and only VP176-immunized mice had complete long-term protection from HSV-2 challenge. PMID- 2550580 TI - Epstein-Barr virus replication in interferon-treated cells. AB - The Burkitt's lymphoma-derived cell line Daudi is latently infected with the Epstein-Barr virus (EBV). These cells are very sensitive to the growth inhibitory and differentiation-inducing effects of human interferon-alpha (IFN) and we have examined the possibility that these responses are due to induction of EBV replication. Our results indicate that only a very small proportion of cells (less than 0.3%) are induced by IFN treatment to express EBV lytic genes at the RNA or protein level, whereas cell growth inhibition is complete under the same conditions. In contrast, when EBV replication is chemically induced the large increase in lytic gene transcripts, including that of the BZLF1 trans-activator gene, is partially inhibited by concomitant IFN treatment. The increase in viral DNA copy number in chemically induced Daudi cells is also partially inhibited by IFN but no effect of IFN on the level of viral DNA is observed in uninduced cells. PMID- 2550581 TI - Isolation and characterization of temperature-sensitive mutants of Abelson murine leukaemia virus that exhibit dissociation among morphological transformation, soft agar colony-forming ability and tyrosine kinase activity. AB - Seven temperature-sensitive (ts) mutants of Abelson murine leukaemia virus (A MuLV) were isolated on the basis of the temperature dependence of their soft agar colony-forming ability. These seven ts mutants exhibited similar characteristics and were not ts for morphological transformation and autophosphorylation of P120gag-abl protein. The dissociation of the properties of morphology, soft agar colony formation and tyrosine kinase activity might suggest that the v-abl product has more than one primary intracellular target. PMID- 2550582 TI - Enhanced endothelial cytopathogenicity induced by a cytomegalovirus strain propagated in endothelial cells. AB - Endothelial involvement has been implicated in cytomegalovirus (CMV) infection, a source of major complications in immunosuppressed individuals (e.g., those with acquired immune deficiency syndrome [AIDS] and organ transplants). Traditionally, CMV has been grown in fibroblasts; however, propagation in these cells may alter characteristics of the virus. In developing an in vitro model system of CMV/endothelial cell interaction, we have addressed this issue by propagating a clinical isolate, CMV VHL 1, in human umbilical vein endothelial (HUVE) cells by serial cocultivation of heavily infected cultures with fresh HUVE monolayers and have compared its infectious properties with those of the fibroblast-raised strain, CMV AD169. In situ hybridization using a biotinylated DNA probe, as well as immunofluorescent staining for CMV-specific antigen, has confirmed infection of HUVE cells inoculated with either strain of the virus. Infection of HUVE by VHL was accompanied by dramatic cytopathology not observed in AD169-infected cells. Plaque assay of culture supernatants revealed greater virus production in VHL-infected HUVE as compared with equivalently inoculated fibroblasts. In contrast, AD169 production in inoculated fibroblasts exceeded that in HUVE. These studies demonstrate the suitability of cultured endothelial cells as a substrate for CMV propagation and suggest that a strain of virus thus propagated may offer an accurate model of CMV/endothelial cell interaction in human disease. PMID- 2550583 TI - Immunity in Pekin ducks experimentally and naturally infected with duck hepatitis B virus. AB - The immune response to duck hepatitis B virus (DHBV) had not been elucidated. An assay was therefore established to detect the presence of antibody to DHB surface antigen (anti-DHBs) in serum of experimentally inoculated and naturally infected ducks. Anti-DHBs in serum was detected by indirect RIA from the percentage inhibition of binding of rabbit anti-DHBs to purified DHBsAg. Specificity was confirmed by positive and negative controls, infected and noninfected sera, and a mouse monoclonal antibody to DHB core antigen (anti-DHBc). Serum and liver samples were tested for DHBV DNA by dot-blot hybridization assay. Adult ducks repeatedly inoculated with DHBV remained non-viraemic but developed anti-DHBs. This antibody activity neutralized the infectivity of DHBV, which was experimentally inoculated into 1-day-old ducklings. In naturally infected flocks anti-DHBs was detected in a proportion of noninfected adult ducks as well as 1 day-old hatchlings. Anti-DHBs activity in hatchlings neutralized the infectivity of experimentally inoculated DHBV. Pekin ducks can therefore mount a neutralizing antibody response to DHBV, and immunity may be transferred in ovo from dam to off spring. PMID- 2550584 TI - Detection of human papillomavirus deoxyribonucleic acid by filter in situ hybridization during pregnancy. AB - Samples taken from 101 healthy pregnant women (49 over and 52 under the 20-week gestational period) and 108 healthy nonpregnant women were tested for human papillomavirus (HPV) types. Using 6, 11, 16, and 18 HPV DNA probes, 3-5 x 10(5) exfoliated cells scraped from the cervix were tested by filter in situ hybridization (FISH). Thirty-five of the pregnant women (34.6%) had evidence of the presence of HPV DNA: with 11.8% (12/101) HPV 6; 7.9% (8/101) HPV 11; 8.9% (9/101) HPV 16; and 5.9% (6/101) HPV 18 positivity. HPV DNA was detected in 20.4% (22/108) of the non-pregnant women. Compared with the healthy, nonpregnant group, the higher level of asymptomatic cervical HPV infection was mainly due to the accumulation of HPV 16 and 18 nucleic acids during the gestational period: with detection of HPV 16 in 8/49 cases (16.3%) and of HPV 18 DNA sequences in 4/49 (7.6%) cases. Screening 6-8 weeks after delivery indicated a decline of HPV positivity. Of the 4/12 HPV type 16 positive mothers, only one retained the presence of HPV 16 DNA, whereas neither of the 2/12 type 18 positive women reacted after birth with the type 18 radioactive probe. PMID- 2550586 TI - Detection of hepatitis A virus RNA in serum from patients with acute hepatitis. AB - Hepatitis A virus (HAV) RNA was extracted from the sera of patients with acute hepatitis and then detected by molecular hybridization using cloned HAV complementary DNA (cDNA). HAV RNA was detected in 20 of 85 patients with acute HAV infection, mainly during the prodromal stage, or early during the icteric phase of the disease; it was detected as long as 21 days after its initial detection. Patients with HAV RNA in the serum had a significantly higher titer of anti-HAV IgM. PMID- 2550585 TI - Diagnosis of hepatitis A and B by testing saliva. AB - The use of salivary samples to diagnose acute viral hepatitis was investigated. Tests for IgM antibody to hepatitis A virus (anti-HAV) on 29 acute-phase samples from serologically confirmed cases of hepatitis A were strongly reactive. Follow up samples indicated that IgM anti-HAV persisted at moderate levels for 2-4 months and was not usually detectable thereafter. The ratio of IgM to IgG anti HAV (RIA index) correlated closely with the interval from onset of infection. Significant levels of IgM anti-HAV were not detected in the saliva of 103 IgG anti-HAV positive and 102 IgG anti-HAV negative individuals nor of 4 individuals with hepatitis B. Similarly, IgM anti-HBc was present in the saliva of acute cases of hepatitis B, but not in the saliva of 25 IgG anti-HBc positive and 85 IgG anti-HBc negative individuals, nor of 24 individuals with recent hepatitis A. It is concluded that saliva is a convenient and satisfactory alternative to serum for the diagnosis of hepatitis A infection. PMID- 2550587 TI - Antitumor agents, 107. New cytotoxic 4-alkylamino analogues of 4'-demethyl epipodophyllotoxin as inhibitors of human DNA topoisomerase II. AB - A series of analogues of etoposide, the C-4 amino- and alkylamino-substituted 4' demethyl-epipodophyllotoxins, have been synthesized and studied for their activity to inhibit type II human DNA topoisomerase as well as their activity in causing cellular protein-linked DNA breakage. Substitution of the glycosidic moiety of 1 by a 2"-hydroxyethylamino or 2"-methoxyethylamino chain at the C-4 beta position resulted in potent inhibitors of the human DNA topoisomerase II. This inhibitory activity correlates reasonably well with their activity in causing protein-linked DNA breakage in KB cells. The in vitro cytotoxicity (KB) appears to have no correlation with the inhibitory activity of the human DNA topoisomerase II. PMID- 2550588 TI - Determination of clefts in receptor structures. AB - The automatic determination of atoms which comprise a cleft in a receptor is of great importance in computer-aided drug design. X-ray studies of ligand/receptor pairs show that the ligand is often located in a cleft so that this structural feature will indicate a putative binding site. This information can be used in the design of new drugs by database searching and by automatic structure generation. The methods presented in this paper will find the complete accessible surface in a slice through a receptor and also all the clefts and dimples in this surface, using the properties of the Voronoi tessellation of the receptor. Clefts and binding sites can now be determined quickly and without observer bias. PMID- 2550589 TI - Neuroendocrine differentiation: a prognostic feature of non-small-cell lung cancer? PMID- 2550591 TI - Inability of computed tomography appearance of recurrent malignant astrocytoma to predict survival following reoperation. AB - Computed tomographic (CT) scans of 39 patients who underwent reoperation for recurrent malignant astrocytoma at Memorial Sloan-Kettering Cancer Center from 1980 through 1987 were reviewed and correlated with the patients' clinical course. Histologic diagnosis (anaplastic astrocytoma v glioblastoma multiforme) had a statistically significant impact on survival following reoperation (P = .038). Patients with high preoperative performance status (P = .29), total resection by postoperative CT scan (P = .15), and frontal lobe tumors (P = .17) tended to survive longer following reoperation. The size of the tumor at the time of recurrence did not correlate with survival following reoperation. Patients with a small amount of peritumoral edema at the time of recurrence tended to survive longer, but the effect was small (P = .16). Prognosis following reoperation cannot be accurately predicted on the basis of tumor appearance on CT scan. PMID- 2550590 TI - The use of neuroendocrine immunoperoxidase markers to predict chemotherapy response in patients with non-small-cell lung cancer. AB - Small-cell lung cancer (SCLC), a chemotherapy-responsive disease, is characterized by neuroendocrine properties. In contrast, non-small-cell lung cancer (NSCLC) is at best moderately responsive to chemotherapy, and only 10% to 20% of cases demonstrate neuroendocrine properties. The present study is a retrospective analysis of the use of immunoperoxidase markers for neuron-specific enolase (NSE), Leu-7, and chromogranin A in NSCLC patients treated with chemotherapy. It was designed to determine if the presence of neuroendocrine markers predict for response to chemotherapy. The diagnostic slides and blocks were obtained on 52 NSCLC patients who were treated with chemotherapy (26 responders and 26 nonresponders). Immunoperoxidase studies were performed, and slides were scored without knowledge of the patient's response. Markers were positive in responders and nonresponders, respectively, as follows: NSE, 14 of 26 (54%) versus seven of 26 (27%), P = .04; Leu-7, 11 of 25 (44%) versus five of 26 (19%), P = .08; and chromogranin A, three of 26 (12%) versus 0 of 26 (0%), P = .71. Two markers were positive in 10 of 26 responders (38%) and 0 of 26 nonresponders (0%), P less than .01. Responders with two or more positive markers showed superior survival (median, 79 weeks) compared with responders with fewer than two positive markers (median, 51 weeks) and nonresponders (median, 27 weeks). These data suggest that the presence of neuroendocrine markers in NSCLC is associated with an increased likelihood of response to chemotherapy and may add to the standard parameters (performance status, weight loss) used to select patients for chemotherapy. PMID- 2550592 TI - Large dose fraction radiotherapy in the treatment of glioblastoma. AB - Twenty-four adults with glioblastoma multiforme (astrocytoma, grade 4) underwent postoperative large dose fraction radiotherapy (LDFR; 5 Gy twice weekly) with Linac X-rays. The outcome in this group was compared with that of 26 patients who received conventional fractionated radiotherapy (CFR; 2 Gy 5 times weekly). The time, dose, and fractionation (TDF) factor was about 100 in both groups. The survival rates following LDFR and CFR were, respectively, 63% vs 65% at 1 year; 36% vs 8% at 2 years; 17% vs 4% at 3 years; and 4% vs 0% at 5 years. Although the survival curve for LDFR was superior to that for CFR, the difference was not statistically significant. Autopsies of nine LDFR and 13 CFR patients showed no residual tumor in one case and no cases, respectively; small residual tumor in three cases in each group; extensive coagulation necrosis of the tumor and surrounding brain tissue in one LDFR and four CFR patients; tumor proliferation in three LDFR and four CFR cases; and mixed glioblastoma and fibrosarcoma in one LDFR and two CFR patients. These results suggest that maximum tumor removal followed by LDFR may offer a better prognosis for patients with glioblastoma than that offered by surgery plus CFR. PMID- 2550593 TI - Variability in the proliferative potential of human gliomas. AB - One hundred forty-three patients with gliomas of astrocytic origin (61 with glioblastomas multiforme (GM) and 82 with astrocytomas) received an intravenous infusion of bromodeoxyuridine (BUdR), 150-200 mg/m2 at the time of surgery, to label tumor cells undergoing DNA synthesis. BUdR-labeled cells were identified by the indirect immunoperoxidase method using anti-BUdR monoclonal antibodies. The percentage of BUdR-labeled cells, or BUdR labeling index (LI), was calculated by microscopic examination of selected viable areas of the tissue sections. The GMs had a median LI of 7.5%, and three quarters of these tumors had LIs greater than 5%. Highly anaplastic astrocytomas (HAAs) and moderately anaplastic astrocytomas (MoAAs) had median LIs of 2.3% and less than 1%, respectively. Among the HAAs, the LI was 1% to 5% in 56% of tumors, greater than 5% in 26%, and less than 1% in 18%. More than 60% of MoAAs had LIs less than 1%, which agrees with the slow clinical progression of this type of tumor, and the remainder had LIs of 1.4% to 9.3%. These results show that histopathologically similar tumors may have different proliferative potentials. Measuring the proliferative potential of individual gliomas is therefore crucial for predicting the prognosis more accurately and for devising more tumor-specific treatment regimens for individual patients. PMID- 2550595 TI - Primary salivary gland tumors. A report of 703 cases. AB - Seven hundred-three cases of primary epithelial tumors of the salivary glands diagnosed over a long period in two Turkish University Hospitals are presented. The patient age distribution, sex, and the locations and histological types of the tumors are discussed and compared with those in the literature. PMID- 2550594 TI - The effectiveness of chemotherapy for treatment of high grade astrocytoma in children: results of a randomized trial. A report from the Childrens Cancer Study Group. AB - Fifty-eight patients with high-grade astrocytoma were treated by members of the Childrens Cancer Study Group in a prospective randomized trial designed to study the effectiveness of chemotherapy as an adjuvant to standard surgical treatment and radiotherapy. Following surgical therapy, patients were assigned randomly to radiotherapy with or without chemotherapy consisting of chloroethyl-cyclohexyl nitrosourea, vincristine, and prednisone. Treatment with chemotherapy prolonged survival and event-free survival. Five-year event-free survival was 46% for patients in the radiotherapy and chemotherapy group, and 18% for patients in the radiotherapy-alone group. Five-year survival was similarly improved. The differences in outcome due to treatment were statistically significant after correcting for imbalances in important prognostic factors (event-free survival, p = 0.026; survival, p = 0.067). The presence of mitoses or necrosis in the tumor specimen was associated with poorer outcome. Patients whose initial surgery was limited to biopsy, and patients with basal ganglia lesions, also had significantly worse outcome. Chemotherapy administered at the time of recurrence in a small number of patients did not produce any long-term survivors. This study is to our knowledge the only randomized trial to investigate effectiveness of chemotherapy in the treatment of high-grade astrocytoma in children. PMID- 2550596 TI - Virology, immunology, and natural history of HIV infection. AB - Human immunodeficiency virus-1 infects and damages or destroys several types of cells, most importantly helper/inducer (CD4+) lymphocytes. In the majority of infected persons, the loss of CD4+ lymphocytes leads to a progressive reduction in both cell mediated and antibody mediated immunity. Early during infection most adults are asymptomatic, but after several years many develop symptoms representing a moderate degree of immune suppression. Eventually, most of these individuals become susceptible to the life-threatening opportunistic infections and cancers which define the acquired immunodeficiency syndrome. Recent advances in management, including earlier diagnosis, the use of maintenance or suppressive therapies, and specific anti-retroviral therapy with zidovudine (AZT) has nearly doubled the life expectancy for persons with AIDS. Clinical trials of new drugs, sometimes in combination with AZT, are underway. Vaccine development is proceeding, but many obstacles must be overcome before a successful vaccine can be identified. PMID- 2550597 TI - Interaction of bile acids, phospholipids, cholesterol and triglyceride with dietary fibers in the small intestine of rats. AB - Certain dietary fibers have been reported to lower plasma cholesterol by binding bile acids and reducing their recycling through the enterohepatic circulation. In addition, certain fibers may delay the digestion and absorption of fat. In the present study, the interaction of bile acids with guar gum (GG), konjac mannan (KM) and chitosan (CH) was determined. Rats were fed during a 20-min period a test meal containing either 5% cellulose (CE), GG, KM or CH and also containing 14C-labeled triolein and 3H-labeled cholesterol. The group fed CE served as control, since CE does not bind bile acids or phospholipids in vivo. Two hours after presentation of the test meal, rats were killed and the stomach and small and large intestine removed. All four groups ate the same amount of the test meal, about 1.9 g. The aqueous phase of the small intestinal contents was separated by ultracentrifugation, and the amount (mumol) of bile acids and phospholipids in the total intestinal contents and in the aqueous phase was estimated. The ratio of bile acids in the aqueous phase to that in total intestinal contents was significantly higher in the GG and KM groups and significantly lower in the CH group than that in the CE group, demonstrating that the bile acids are bound or trapped by each of these fiber sources. Only CH appeared to bind phospholipids, reducing the proportion in the aqueous phase compared to that in the CE group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550598 TI - Fermentable dietary fibers elevate urinary methylmalonate and decrease propionate oxidation in rats deprived of vitamin B-12. AB - This study examines the effect of dietary fiber supplements of different degrees of bacterial fermentability on biochemical indicators of vitamin B-12 deficiency in rats. Groups of rats were fed a fiber-free diet deficient in vitamin B-12 or the fiber-free diet diluted with 5% of a poorly fermentable dietary fiber (cellulose, lignin or alginic acid) or a highly fermentable fiber (pectin, guar gum or xylan). Poorly fermentable fibers had no significant effect on apparent B 12 status, whereas the highly fermentable fibers significantly increased urinary methylmalonic acid and depressed oxidation of [14C]propionate to 14CO2. Pectin consistently induced significantly greater effects than did xylan or guar gum. The data are consistent with the hypothesis that fermentable fibers stimulate bacterial propionate production and exaggerate certain biochemical indicators of B-12 deficiency. Since pectin had a more pronounced effect than did other fermentable fibers, the possibility that pectin may also interfere with B-12 absorption requires further study. PMID- 2550599 TI - Dietary pectin shortens the biologic half-life of vitamin B-12 in rats by increasing fecal and urinary losses. AB - As little as 5% of pectin added to a fiber-free diet elevates urinary methylmalonic acid (MMA) severalfold in vitamin B-12--deprived rats. The present study examines whether increased urinary MMA reflects lower vitamin B-12 status or occurs only because of fermentation of pectin by intestinal bacteria and increased production of propionate, a precursor of MMA. By monitoring urinary and fecal excretion of 57Co after a tracer dose of [57Co]vitamin B-12, we found the biologic half-life of vitamin B-12 to be 59 d for rats fed a fiber-free diet and only 19 d for rats fed a 5% pectin diet. Also, pectin-fed rats oxidized only 12% of a 1-mmol dose of [14C]propionate to 14CO2 in 2 h, whereas rats fed the fiber free diet expired 33% of the dose. Finally, high urinary MMA persisted even after the removal of pectin from the diet. We conclude that dietary pectin accelerates vitamin B-12 depletion in rats, possibly by interfering with enterohepatic recycling of vitamin B-12. By stimulating microbial propionate production, pectin and other fermentable fibers may also contribute to increased urinary MMA in vitamin B-12 deficiency, but a larger propionate pool does not account for the other effects of pectin on vitamin B-12 status. PMID- 2550600 TI - ras oncogene p21 expression in hepatocellular carcinoma. AB - Aberrant proto-oncogene expression has been implicated in hepatic cell proliferation, transformation and carcinogenesis using a rat model. To investigate the role of ras p21 product expression in human hepatocellular carcinoma (HCC), we have localized ras p21 in formalin fixed, paraffin-embedded normal and abnormal livers utilizing the avidin-biotin peroxidase method and a monoclonal antibody to ras-gene product p21. A semi-quantitative estimate of p21 expression was performed by serial dilutions of primary antibody. While low dilutions of anti-p21 stained normal hepatocytes, higher dilutions failed to react with normal hepatocytes and these dilutions were used for assessment of p21 enhancement. Increased p21 expression of ras oncogene in HCC occurs in fibrolamellar carcinomas and other better differentiated HCC. Tumor dedifferentiation is associated with an attenuation of p21 expression. Liver adjacent to HCC exhibits p21 enhancement, in contrast to liver surrounding metastatic carcinoma, suggesting increased p21 expression in HCC induction. PMID- 2550601 TI - Hepatic disease associated with intrauterine parvovirus B19 infection in a newborn premature infant. AB - A hydropic premature infant with intrauterine growth retardation died at 4 days of age and was found at necropsy to have advanced liver disease. Clinical and serologic findings in mother and infant were consistent with recent parvovirus B19 infection. Parvovirus can cause fetal liver disease in animals, and some instances of congenital hepatic dysfunction in man may be due to intrauterine parvovirus B19 infection. PMID- 2550602 TI - Development of acid secretory function in the rat stomach: sensitivity to secretagogues and corticosterone. AB - Gastric acid secretion was studied in anesthetized rats from day 6 of the postnatal period up to the time of weaning. Basal H+ secretion was detected from day 6 in the first group studied (2.4 +/- 0.2 muEq of H+/10 min/100 g of body weight, BW) and remained constant up to the time of weaning (day 18: 2.5 +/- 0.2 muEq of H+/10 min/100 g of body weight) except for the period between days 10 and 12, when it fell significantly (1.5 +/- 0.06 muEq H+/10 min/100 g of BW on day 12). Both histamine H2 receptor sensitivity and intracellular transduction mechanism activities were evaluated by studying the secretory responses to histamine, impromidine (an H2 receptor agonist), cimetidine (an H2 receptor antagonist), forskolin (a direct adenylate cyclase activator), and dibutyryl (db) cAMP (an analogue of cAMP, the intracellular messenger mediating the response to histamine). The effects of pentagastrin and carbachol were also determined. The secretory responses obtained on days 6, 8, and 18 were similar and represented about threefold increases over basal secretion for all the secretagogues used. After weaning on day 20, both the basal secretion and the response to secretagogues were significantly increased compared with those of unweaned animals. On day 12, the responses were always weaker than on both days 8 and 18. Injection of 1 mg/kg of corticosterone 21 acetate daily from day 8 resulted on day 12 in a basal secretion and a response to histamine equivalent to those measured in 18-day-old pups not injected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550603 TI - Resection of a tumor of the psoas muscle. AB - A 16-year-old boy presented with paresthesias and weakness of the right thigh. A computerized axial tomography scan identified a mass within the right psoas muscle, and a needle biopsy showed a primitive neuroectodermal tumor, which did not respond to two courses of chemotherapy (Adriamycin, vincristine, and cytoxan). An anatomic resection of the psoas muscle and femoral nerve was performed and radiotherapy administered. The boy remains ambulatory and free of disease 2.5 years after resection. PMID- 2550604 TI - Gingival fibroblasts degrade type I collagen films when stimulated with tumor necrosis factor and interleukin 1: evidence that breakdown is mediated by metalloproteinases. AB - We previously suggested that periodontal pathogens might mediate connective tissue degradation in periodontal diseases through the ability of antigens from their cell walls to stimulate cytokine production by circulating mononuclear cells. Such cytokines would then induce metalloproteinase (MP) synthesis by resident gingival cells and thus initiate matrix degradation. In the present investigation human gingival fibroblasts (HGFs) were grown on [14C]-labelled type I collagen films and stimulated with either tumor necrosis factor (TNF) or interleukin-1 (IL-1) for 48 h. Collagenolysis occurred in a dose-dependent manner; the optimal dose for human rTNF alpha was 100 ng/ml and for rIL-1 alpha and rIL-1 beta, 1 ng/ml. Collagen degradation was accompanied by increased synthesis and release of the MPs collagenase, gelatinase and stromelysin, and there was a reduction in free TIMP (tissue inhibitor of metalloproteinases): collagenase and stromelysin were detected in both active and latent forms. Cytokine-stimulated collagenolysis was abolished by the addition of exogenous human rTIMP (5 units/ml). We also measured collagenase and TIMP by ELISAs which recognize all forms of collagenase (latent, active or complexed) and TIMP (free or complexed). These showed that while collagenase activity (0.6-1.2 microgram/ml) correlated with lysis, total TIMP levels remained unchanged at approximately 0.2 microgram/ml. These results demonstrate important roles for MPs and TIMP in regulating type I collagen degradation by HGFs, and support the hypothesis that connective tissue destruction during inflammatory diseases may be initiated, at least in part, by TNF and IL-1. PMID- 2550605 TI - Repopulation of dentin surfaces by periodontal ligament cells and endothelial cells. Effect of basic fibroblast growth factor. AB - The regeneration of connective tissue attachment is a major goal of clinical periodontics. Recent investigations on biochemically mediated periodontal regeneration have attempted to define the various biological response modifiers which may provide a mechanism for periodontal regeneration. Fibronectin and endothelial cell growth factor have been shown to selectively enhance periodontal ligament (PDL) cell adhesion, migration, and proliferation. In addition, dentin preconditioned with tetracycline HCl (TTC) or citric acid (CA) supports PDL cell adhesion, presumably by exposing collagen fibers. We have now extended these studies to include basic fibroblast growth factor (b-FGF) as a potential meditor of periodontal regeneration. Using AFSCM (assays for specific cell migration), b FGF in concentrations as low as 10 ng per dentin block significantly stimulated PDL cell chemotaxis, while the antibody against b-FGF inhibited both the chemotactic and proliferative characteristics of the mitogen. We also found that 5 ng and above of b-FGF per dentin block significantly stimulated human endothelial cell migration and proliferation. Using 125I-b-FGF, we demonstrated that the factor binds to native dentin. This binding was increased when the dentin blocks were preconditioned by TTC or CA and reduced when the dentin was subsequently treated with collagenase. 125I-b-FGF also bound with moderate affinity to a type I collagen affinity column whereas the binding to a hydroxylapatite affinity column was negligible. The combination of FN and b-FGF was a marginally more potent chemo-attractant than b-FGF alone for PDL cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550606 TI - Potentiation of therapeutic effect of methanesulphonate and protection against its organ cytotoxicity by vitamin C in Ehrlich ascites carcinoma bearing mice. AB - The effect of vitamin C upon the therapeutic index and side effects produced by methanesulfonate of aminoglycols (drug 864T, NSC 140117) had been evaluated in a laboratory system. The antitumor action of 864T, vitamin C and their combination were evaluated in Ehrlich ascites carcinoma (EAC) cells in vivo. Tissue toxicity was assessed using liver and intestinal DNA, RNA, protein contents and their synthesis as parameters. In addition, G-6-pase, 5-Nase and Alk, pase activity levels in both tissues were also measured. Drug 864T (200 mg/kg) produced 50 percent long-term survivors in tumor bearing mice in addition to 10 percent early mortality while in combination with vitamin C (250 mg/kg x 6), there was 80 percent long term survivors with no mortality related to drug toxicity. No toxicity, in all the parameters used, was observed when 864T was given in combination with vitamin C. Drug 864T alone produced a significant decrease in protein content of both liver and intestinal tissue while in combination with vitamin C normal levels were maintained. In addition, all the parameters used were either elevated or decreased by 864T treatment and returned to normal levels in combination with vitamin C. This study proved that vitamin C may be useful not only to potentiate the effect of anticancer drug 864T on the Ehrlich ascites carcinoma but also to antagonize the side effects of the drug. PMID- 2550607 TI - Immunochemical studies for the presence of P-450 HFLa, a form of cytochrome P-450 in human fetal livers in human hepatocellular carcinoma cells. AB - Immunohistochemical localization of P-450 HFLa, a form of cytochrome P-450 in human fetal livers was investigated in human hepatocellular carcinoma. The cytoplasm of carcinoma cells positively reacted with anti-P-450 HFLa antibodies. It was found that the carcinoma cells showing a pseudoglandular pattern or poorly differentiated appearance exhibited a weaker reactivity with anti-P-450 HFLa antibodies than did relatively differentiated carcinoma cells. In the case of hepatoblastoma, the polygonal or round-shaped tumor cells which differentiated into epithelial structure exhibited a positive reaction with anti-P-450 HFLa antibodies, whereas the spindle-shaped tumor cells which showed a sarcomatous appearance did not react with the antibodies. PMID- 2550608 TI - Cytotoxic effect of 8-amino adenosine 3',5'-cyclic monophosphate on FM3A mouse mammary tumor cells. AB - Derivatives of adenosine 3',5'-cyclic monophosphate (cAMP) with modifications at the 8-position were synthesized and examined for their cytotoxic effects on FM3A mouse mammary tumor cells and ZR-75 human mammary tumor cells. On in vitro tests of these derivatives, 8-amino (8-NH2) cAMP was the most effective against both cell lines. This compound showed the dose-dependent inhibition of FM3A cell growth in the concentration of over 2.5 microM with the ID50 value of 4 microM. Furthermore, antitumor activity of 8-NH2 cAMP was also tested against FM3A cells in vivo. T/C% values of 8-NH2 cAMP were respectively 162% and 138% in response to doses of 30 and 10 mg/kg by intraperitoneal injections of 8-NH2 cAMP for 5 d. 8 NH2 cAMP was converted to 8-NH2 adenosine via 8-NH2 adenosine 5'-monophosphate by some enzymes in the fetal bovine serum and the cytotoxic effect of 8-NH2 cAMP on FM3A cells was actually stemmed from 8-NH2 adenosine. PMID- 2550609 TI - Degradation and disposal of some antineoplastic drugs. AB - Bulk quantities and pharmaceutical preparations of the antineoplastic drugs carmustine (BCNU), lomustine (CCNU), chlorozotocin, N-[2-chloroethyl]-N'-[2,6 dioxo-3-piperidinyl]-N-nitrosourea (PCNU), methyl CCNU, mechlorethamine, melphalan, chlorambucil, cyclophosphamide, ifosfamide, uracil mustard, and spiromustine may be degraded using nickel-aluminum alloy in KOH solution. The drugs are completely destroyed and only nonmutagenic reaction mixtures are produced. Destruction of cyclophosphamide in tablets requires refluxing in HCl before the nickel-aluminum alloy reduction. Streptozotocin, chlorambucil, and mechlorethamine may be degraded using an excess of saturated sodium bicarbonate solution. The nitrosourea drugs BCNU, CCNU, chlorozotocin, PCNU, methyl CCNU, and streptozotocin were also degraded using hydrogen bromide in glacial acetic acid. The drugs were completely destroyed but some of the reaction mixtures were mutagenic and the products were found to be, in some instances, the corresponding mutagenic, denitrosated compounds. PMID- 2550610 TI - Toothpick foreign body in the foot. A case report. AB - A case report of a wooden toothpick as a foreign body in the foot was presented. Such lesions are visualized poorly, if at all, on standard radiographs, and localization of the foreign body may not be possible. Computed tomography, with its superior soft tissue imaging capabilities, can detect this otherwise radiolucent wood material and make accurate localization of the foreign body possible. PMID- 2550611 TI - Stereospecificity of the discriminative stimulus functions of the dimethylheptyl homologs of 11-hydroxy-delta 8-tetrahydrocannabinol in rats and pigeons. AB - Rats and pigeons were trained to discriminate between the presence and absence of the effects of (-)-delta 9-tetrahydrocannabinol (THC) at doses of 3 and 0.56 mg/kg, respectively; injections were i.p. and i.m., 0.5 and 1.5 hr before session onset for the two species, respectively. Tests with the 1,1 dimethylheptyl (DMH) homolog of (-)- delta 8-THC as well as its 11-hydroxylated (11-OH) derivative [( )-11-OH-delta 8-THC-DMH], showed that both compounds were more potent than the training compound, especially so in the case of the 11-OH product (66 and 80 times more potent than delta 9-THC in rats and pigeons, respectively). The enantiomer, (+)-11-OH-delta 8-THC-DMH, was inactive as a THC-like psychotomimetic in doses even up to 10 mg/kg [ED50 of (-)-11-OH-delta 8-THC being 0.01 mg/kg and 0.002 mg/kg in rats and pigeons, respectively]. Hence, the typical THC-like effects in rats and pigeons (and by extrapolation possibly also the feeling of "high" in humans), reside exclusively in the levo [(-)]-enantiomers of THC-type cannabinoids (at least in the pair studied by us) as demonstrated clearly in this study. Both (-)-delta 8-THC-DMH and (-)-11-OH-delta 8-THC-DMH had a slow onset and a long duration of action. PMID- 2550613 TI - Obesity modifies the adrenergic status of dog adipose tissue. AB - The relative proportion of antilipolytic alpha-2 and lipolytic beta adrenoceptors and the adrenoceptors was studied in adipocytes from lean and obese dogs. The modification of the adrenergic status in the adipose tissue from obese dogs consists of an increase in alpha-2 adrenoceptor number (identified by [3H]yohimbine) and a decrease in beta adrenoceptor number (identified by [3H]dihydroalprenolol). Neither the number of beta adrenoceptors in the leukocytes nor the number of alpha-2 adrenoceptors in the platelets were altered in obesity. This predominance of alpha-2 adrenoceptors in adipocytes from obese dogs induced a reduction of the lipolytic efficacy of epinephrine (i.e., increase in the concentration able to induce half-maximal stimulation of lipolysis). Moreover, the number of beta adrenoceptors in the high-affinity state was increased in adipose tissue from obese dogs. It is concluded first that the striking modifications in the adrenergic status of the adipose tissue in obesity is specific to this tissue and secondly that the rise of the beta adrenoceptor in the high-affinity state could explain the fact that catecholamines remain lipolytic agents and that weight loss is increased by starvation in the obese dog. PMID- 2550612 TI - Central and peripheral opioid modulation of gastric relaxation induced by feeding in dogs. AB - The influence of i.v. vs. i.c.v. administration of [( D-Ala2, MetPhe4, Gly ol5]enkephalin (DAMGO) and morphine), kappa U 50488 and ethylketocyclazocine and delta ([D-Pen2, D-Pen5]enkephalin (DPDPE)] opioid agonists on gastric relaxation induced by a standard meal was evaluated in conscious dogs with strain-gauge transducers implanted on the gastric fundus and antrum. Under control conditions, the amplitude of the gastric relaxation in response to feeding was 2.46 +/- 0.23 g. Given i.v. 10 min before feeding, both U 50488 (10 micrograms/kg) and ethylketocyclazocine (10 micrograms/kg) significantly (P less than .01) reduced the amplitude of the gastric relaxation by 57 and 68%, respectively, whereas DAMGO and morphine (10 micrograms/kg i.v.) increased markedly the response to feeding by 67 and 51%, respectively. In contrast, DPDPE had no effect on the gastric relaxation induced by feeding. Previous administration of naloxone (0.3 mg/kg i.v.) or MR 2266 (0.3 mg/kg i.v.) blocked the effect of U 50488 on gastric relaxation with no effect per se on the amplitude of response; naloxone also blocked the increase in gastric relaxation induced by DAMGO. When administered i.c.v. (0.1 microgram/kg) DAMGO induced a significant (P less than .05) increase in the amplitude of gastric relaxation whereas U 50488 and DPDPE (0.1 and 1 microgram/kg i.c.v.) had no effect. The effect of i.c.v. DAMGO on gastric relaxation was unaffected by a previous i.v. administration of SR 58002C (1 mg/kg). Truncal vagotomy blocked the increase in gastric relaxation induced by DAMGO (10 micrograms/kg i.v.), but did not change the effect of U 50488 (10 micrograms/kg i.v.) on gastric relaxation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550614 TI - Effects of opioid agonists on sympathetic and parasympathetic transmission to the dog heart. AB - Effects of leu- and met-enkephalin, pentazocine and morphine on negative or positive chronotropic response to vagal nerve stimulation or cardiac sympathetic nerve stimulation were examined in anesthetized dogs in order to determine whether opioid receptors modulate vagal and sympathetic transmission. Leu- and met-enkephalin (10-100 micrograms/kg i.v.) and pentazocine (100-1000 micrograms/kg i.v.) inhibited bradycardic response to vagal nerve stimulation (1 4 Hz) in a dose-dependent manner. Morphine (300 and 1000 micrograms/kg i.v.) did not affect vagal bradycardia. The inhibitory effect of leu-enkephalin (30 micrograms/kg) and pentazocine (300 micrograms/kg) was effectively antagonized by naloxone (1000 micrograms/kg i.v.). Bradycardic response to intracoronary injection of methacholine (0.1, 0.3 and 1 microgram) into the right coronary artery was unaffected by leu-enkephalin (30 micrograms/kg). On the other hand, leu-enkephalin and pentazocine did not modify tachycardic response to sympathetic nerve stimulation (1-8 Hz). Morphine attenuated sympathetic tachycardia only slightly. These results suggest that presynaptic opioid receptors, probably delta type, are present in the vagus nerves, and that the activation of opioid receptors inhibit vagal transmission to the dog heart. In contrast, the presence of opioid receptors in the cardiac sympathetic nerves is not evident. PMID- 2550615 TI - Inhibition of vagal transmission by cardiac sympathetic nerve stimulation in the dog: possible involvement of opioid receptor. AB - The inhibitory effect of cardiac sympathetic nerve stimulation (SNS) on bradycardic response to vagal nerve stimulation was examined in anesthetized dogs pretreated with atenolol or propranolol to prevent tachycardia caused by SNS or norepinephrine infusion. The amplitudes of vagal bradycardia (0.5, 1, 2, and 4 Hz) during 10 and 30 Hz of SNS were significantly smaller than those during the resting state. The inhibitory effect of 10 Hz of SNS was neither affected by prazosin (10, 30 and 100 micrograms/kg i.v.) nor by yohimbine (10, 30 and 100 micrograms/kg i.v.). The SNS-induced inhibition of vagal bradycardia was inhibited by naloxone (0.3 and 1 mg/kg i.v.) or by naltrexone (0.3 and 1 mg/kg i.v.). Vagal bradycardia was unaffected by intracoronary infusion of norepinephrine (3 micrograms/min) into the right coronary artery, whereas it was effectively inhibited by leu-enkephalin (10, 30 and 100 micrograms/kg i.v.). Bradycardia induced by intracoronary injection of methacholine (0.3, 1 and 3 micrograms) was unaffected by SNS. These results suggest that a presynaptic alpha adrenoceptor mechanism is not involved in the SNS-induced inhibition of vagal bradycardia in the dog, and suggest further that an opioid receptor mechanism may be responsible for the inhibition. PMID- 2550616 TI - Relation between density (maximum binding) of alpha adrenoceptor binding sites and contractile response in four canine vascular tissues. AB - In microsomal fractions from dog aorta, saphenous veins, mesenteric arteries and veins, both [3H]prazosin and [3H]rauwolscine displayed monophasic saturation in binding. The Kd for [3H] rauwolscine binding was similar for all these blood vessels, but the maximum number of [3H]rauwolscine binding sites was 3 to 7 times higher in veins compared to arteries. The Kd for [3H] prazosin was higher in saphenous vein than that in the arteries. The maximum number of binding sites for [3H]prazosin was similar, except for that in aorta, which was 3 times greater. Phenylephrine (alpha-1 adrenoceptor selective agonist) or norepinephrine (nonselective adrenoceptor agonist) produced similar maximal responses in all vessels. The alpha-2 adrenoceptor selective agonist, B-HT 920 (2-amino-6-allyl 3,4,7,8-tetrahydro-6H-thiazolo[5,4-d]-azepine)-induced contraction in veins but not in arteries. Prazosin (10(-6) M) inhibited completely the contractions to norepinephrine (3 x 10(-6) M) in mesenteric arteries and to phenylephrine (3 x 10(-6) M) in arteries and veins. Contractile responses of mesenteric artery were unaffected by rauwolscine. Rauwolscine (10(-7) M) caused a greater parallel rightward shift of the concentration-response curve to norepinephrine than did prazosin (10(-7) M) in saphenous veins, and a further rightward shift of responses to norepinephrine after 10(-7) M prazosin in mesenteric vein and saphenous vein and abolished B-HT 920-induced responses at alpha-2 adrenoceptors. The tissues responding to B-HT-920 correspond to those having the highest alpha-2 receptor density as measured by [3H]rauwolscine binding. The density of such sites required for contraction to be initiated in veins was much higher than with alpha-1 adrenoceptor sites.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550617 TI - Alpha-1 adrenergic stimulation of 1,4,5-inositol trisphosphate formation in ventricular myocytes. AB - We demonstrated previously that alpha-1 adrenergic catecholamines modulate cardiac automaticity in a manner that is dependent upon the function of a pertussis toxin sensitive guanine nucleotide binding protein (G protein). Furthermore, we demonstrated that alpha-1 adrenergic receptor stimulation promotes the accumulation of inositol monophosphate (IP1). In the present study we used high-pressure liquid chromatography to resolve individual inositol phosphate isomers formed in norepinephrine-stimulated cultured rat ventricular myocytes. Norepinephrine stimulated a rapid, transient increase in 1,4,5-inositol trisphosphate (1,4,5-IP3) which was followed by slower, sustained increases in 1,3,4-IP3, inositol bisphosphate (IP2) and IP1. IP1 was composed of two major isomers with retention times characteristic of 1-IP1 and 4-IP1. 4-IP1 was the predominant IP1 isomer formed during stimulation with norepinephrine suggesting that the polyphosphoinositides rather than phosphatidylinositol are the principal targets of norepinephrine-stimulated phospholipase C activity in the heart. This was confirmed in studies performed on myocyte membranes which demonstrated proportionately greater IP2 and IP3 (relative to IP1) accumulation in response to norepinephrine. G protein regulation of alpha-1 adrenergic-dependent inositol phospholipid hydrolysis also was examined. In myocyte membranes, guanosine-5'-0 (3-thiotriphosphate) induced the accumulation of IP2 and IP3 and was required for the stimulatory effect of norepinephrine. This response was not impaired after pretreatment with pertussis toxin. These results indicate that the myocyte alpha 1 adrenergic receptor is coupled to a polyphosphoinositide-specific phospholipase C by a pertussis toxin insensitive G protein and suggest that under certain conditions IP3 may serve an important role in alpha-1 adrenergic modulation of cardiac function. PMID- 2550618 TI - Solubilization and characterization of the liver peripheral-type benzodiazepine receptor. AB - The rat liver membrane-bound and digitonin-solubilized peripheral-type benzodiazepine receptors (mPBZR and dsPBZR, respectively) were characterized. Forty percent of the receptors were solubilized from a liver homogenate with 0.25% digitonin. Scatchard analysis of saturation data for the mPBZR and the dsPBZR showed Kd = 1.5 nM and maximum number of binding sites = 3.12 pmol/mg of protein and Kd = 9.2 nM and maximum number of binding sites = 1.10 pmol/mg of protein, respectively. Estimates of Kd calculated from kinetic data agree with estimates from Scatchard analysis. The affinity of the PBZR for [3H]Ro5-4864 was not affected by guanosine 5'0-(3-thiotriphosphate) which suggests the receptor is not coupled to a G-protein. Competition for specific [3H]Ro5-4864 binding by various ligands demonstrated the same rank order potency of binding inhibition for the membrane bound and solubilized receptors (PK-11195 greater than or equal to Ro5-4864 greater than diazepam greater than clonazepam). Thus, the soluble receptor had ligand binding characteristics similar to those of the membrane PBZR. [3H]PK-14105 was used to photoaffinity label the PBZR in a rat liver homogenate. Labeling was specific for the PBZR and the molecular weight of the digitonin-solubilized photoaffinity-labeled receptor was estimated to be 170 kDa by gel filtration chromatography. Estimation of the molecular weight of the [3H]PK-14105 labeled receptor by sodium dodecyl sulfate polyacrylamide gel electrophoresis demonstrated a single protein corresponding to 19 kDa. PMID- 2550620 TI - Antagonism of platelet activating factor receptor binding and stimulated phosphoinositide-specific phospholipase C in rabbit platelets. AB - The objective of this study was to establish whether binding of platelet activating factor (PAF) to its receptor was integral to the stimulation of phosphoinositide-specific phospholipase C (PLC) in rabbit platelets. Saturation binding curves for [3H]PAF indicated that the PAF receptor has a dissociation constant (KD) of 28.72 nM. In comparison, PAF-stimulated PLC activity, as monitored by [3H]inositol triphosphate production, increased at lower concentrations and had an half-maximal effective concentration (EC50) value of 1.5 nM. Unlabeled PAF inhibited [3H]PAF binding competitively and demonstrated two binding sites, a high affinity site with an inhibitory constant (Ki) of 2.65 nM and a low affinity site with a Ki of 0.80 microM. The inhibitory effects of four PAF antagonists, CV-3988, CV-6209, SRI 63-441 and SRI 63-675 on the binding of [3H]PAF were compared to the effects of the antagonists on PAF-stimulated PLC activity. The four antagonists inhibited [3H]PAF binding almost completely whereas their ability to inhibit PAF-stimulated PLC activity varied. CV-3988, SRI 63-441 and SRI 63-675 had half-maximal inhibitory concentration (IC50) values of 0.28, 0.78 and 0.42 microM, respectively, whereas CV-6209 was more potent at inhibiting [3H]PAF binding (IC50 = 7.73 nM). The SRI 63-441 and SRI 63-675 inhibited PLC totally with an IC50 value of 0.78 and 1.27 microM, respectively. The CV-3988 and CV-6209 showed a maximal PLC inhibition of about 45% with "apparent IC50" values of 1.05 and 0.17 microM, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550619 TI - Inhibition of bradykinin-induced increases of cytosolic Ca++ by a novel amiloride analog. AB - The present study was undertaken to determine whether activation of the Na+/H+ antiport regulates the bradykinin (BK)-induced Ca++ transient in Madin-Darby canine kidney cells. An amiloride analog, 5-N-N-ethyl[2-methoxy 5 nitrobenzyl]amiloride (L651,548), led to a dose-dependent inhibition of the BK induced Ca++ transient (Ki = 9 microM), and this effect was eliminated in high external Na+. Zero external Na+ failed to inhibit the BK-induced Ca++ release, suggesting L651,548 is not acting through inhibition of the Na+/H+ antiport. Use of monovalent ionophores to adjust the intracellular pH demonstrated changes in peak Ca++ release by BK only over wide pH ranges. Furthermore, studies revealed L651,548 was much more potent at inhibiting pH recovery (Ki = 55 nM) than BK induced Ca++ release. Studies in saponin-permeabilized cells demonstrated no effect of L651,548 on inositol trisphosphate-induced Ca++ release. Whole cell [3H] BK binding studies indicated a dose-dependent inhibition by L651,548. We conclude that the Na+/H+ antiport does not play a critical role in control of BK induced Ca++ release in Madin-Darby canine kidney cells. Also, the amiloride analog L651,548 reduces BK Ca++ release by inhibition of BK receptor binding in Madin-Darby canine kidney cells. PMID- 2550621 TI - Effect of phthalate acid esters on transport in toad bladder membrane. AB - Sclerosing peritonitis is a serious complication in patients on long-term peritoneal dialysis; it markedly decreases transport of water and solute across the peritoneal membrane. Although the precise mechanism is unknown, organic compounds (i.e., plasticizers) from plastic tubing and dialysis bags have been suggested to be a cause of the syndrome. The effects of three such compounds on water and sodium transport in vitro were studied in the toad bladder. The compounds studied were didodecylphthalate, dioctylphthalate, and benzylbutylphthalate. After 4 hr incubation in vitro, dioctylphthalate and benzylbutylphthalate significantly inhibited vasopressin-stimulated water flow in toad bladder. Basal water flow was not affected by any of the three compounds. Sodium transport, as measured using short-circuit current, was decreased to an equivalent degree by all compounds; inhibition of short-circuit current was dose dependent and was approximately 30% at 10(-3) M. The onset of action was between 3.5 and 4 hr, and the effect on short-circuit current was not reversible. These results demonstrate that the plasticizers (to which patients of all sorts are commonly exposed) inhibit transport across living membranes. In the toad bladder these compounds decrease sodium transport and maximal water flow. Although other evidence suggests that the cumulative toxic effects of these compounds may play a causal role in sclerosing peritonitis in patients on peritoneal dialysis, our study suggests that chronic exposure to the phthalate acid esters in patients with normal renal function may result in sodium wastage, polyuria, and a concentrating defect resistant to AVP. PMID- 2550622 TI - Pharmacological characterization of supersensitivity to naltrexone in squirrel monkeys. AB - Effects of naltrexone and of other drugs in decreasing rates of schedule controlled responding were studied in squirrel monkeys treated weekly with naltrexone while responding under fixed-ratio schedules of either food presentation (FP) or stimulus-shock termination (SST). By the 5th week of treatment, sensitivity to the rate-decreasing effects of naltrexone had increased 32-fold in FP monkeys (supersensitivity) but had not changed in SST monkeys; continued weekly injections of naltrexone did not change the effects of naltrexone further in either group. FP monkeys were more sensitive than SST monkeys to the rate-decreasing effects of other opioid antagonists and mixed agonist-antagonists; however, for some compounds (e.g., MR 2266) differences between groups did not exceed differences reported in nonsensitized monkeys responding under similar schedule conditions. Differences in sensitivity between FP and SST monkeys appeared to be stereospecific; FP monkeys were 28 times more sensitive than SST monkeys to (-)-cyclazocine, but equally sensitive to (+) cyclazocine. Sensitivity to opioid agonists and to some nonopioids varied by less than 3-fold between groups and doses of lithium that decreased responding in FP monkeys had no effect in SST monkeys. Neither acute nor repeated injections of morphine altered the sensitivity of FP or SST monkeys to naltrexone; however, morphine attenuated the rate-decreasing effects of naltrexone in FP monkeys, and naltrexone reversed the rate-decreasing effects of morphine in all monkeys. Super sensitivity to opioid antagonists in squirrel monkeys is behaviorally and pharmacologically selective as well as stereospecific. Although other effects of antagonists might contribute to supersensitivity, opioid antagonistic action appears to be one important component of supersensitivity and cross supersensitivity among opioid antagonists. PMID- 2550623 TI - Discriminative stimulus effects of naltrexone in morphine-treated rhesus monkeys. AB - Three adult, female rhesus monkeys responded under a fixed-ratio 5 schedule of stimulus-shock termination while discriminating between s.c. injections of saline and naltrexone (0.01 mg/kg). In addition, monkeys received daily injections of morphine (1.78 or 3.2 mg/kg) 3 hr before experimental sessions. With increasing doses of naltrexone monkeys switched in a dose-related manner from the saline to the naltrexone lever; complete generalization (greater than 80% responding on the naltrexone lever) occurred in all monkeys at doses of naltrexone larger than 0.0032 mg/kg. Doses of naltrexone that produced responding on the drug lever also produced effects typically observed during opioid withdrawal (e.g., miosis and salivation). Compounds with opioid mu antagonist effects under other conditions (e.g., nalorphine) substituted for naltrexone whereas a wide variety of opioid agonists (e.g., morphine, U-50,488 and butorphanol) as well as nonopioids (e.g., pentobarbital and ketamine) produced responding predominantly on the saline lever. Monkeys also switched to the naltrexone lever in a time-related manner after the daily injection of morphine with complete generalization occurring between 8 and 27 hr after morphine. Among a variety of opioid and nonopioid compounds, including drugs used in the treatment of opioid abuse (e.g., clonidine), only compounds with mu agonist actions under other conditions produced a switch in responding from the naltrexone to the saline lever in 27-hr morphine-abstinent monkeys. These results suggest single, daily injections of morphine are sufficient to produce dependence in rhesus monkeys and indicate further drug discrimination studies in morphine-treated rhesus monkeys might be pharmacologically more specific than observational procedures. PMID- 2550624 TI - Comparison of contractions to serotonin, carbamylcholine and prostaglandin F2 alpha in rat stomach fundus. AB - Contractile effects of serotonin were compared to those of carbamylcholine and prostaglandin (PG) F2 alpha in an effort to characterize serotonergic receptor activation in rat stomach fundus. All three agents elicited concentration dependent contractions of fundal strips with serotonin (EC50 = 2 X 10(-10) M) being approximately 100-fold more potent than carbamylcholine (EC50 = 2 X 10(-8) M) and PGF2, (EC50 = 10(-8) M). The calcium channel blockers, diltiazem (5 X 10( 7) to 5 X 10(-5) M) and nitrendipine (10(-8) to 10(-5) M), attenuated responses markedly to serotonin and PGF2 alpha whereas having only minimal effects on carbamylcholine-induced contractions. Neither serotonin (10(-11) to 10(-5) M) nor PGF2 alpha (10(-9) to 10(-5) M) altered [3H]inositol monophosphate generation in fundus whereas carbamylcholine (10(-6) to 10(-5) M) increased significantly [3H]inositol monophosphate with 10(-5) M eliciting an 8-fold increase. Strips of fundus contracted submaximally with either serotonin, PGF2 alpha or carbamylcholine were compared for sensitivity to relaxants. Pinacidil (10(-8) to 10(-5) M) was equipotent (EC50 = 0.1 microM) in relaxing serotonin- and PGF2 alpha-contracted tissues. In contrast, pinacidil was 10-fold less potent in relaxing contractions produced by carbamylcholine. Likewise, nitroglycerin (10( 8) to 10(-4) M) and isoproterenol (10(-12) to 10(-7) M) were at least 10-fold more potent in relaxing serotonin- and PGF2 alpha- than carbamylcholine-induced contractions. Thus, in rat fundus, contractions associated with increased phosphoinositide hydrolysis may be more resistant to relaxation than are contractions not associated with altered phosphoinositide hydrolysis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550625 TI - Diuretic effect of bremazocine, a kappa-opioid with central and peripheral sites of action. AB - Intracerebroventricular or i.p. injections of bremazocine produced a dose dependent diuretic response and increased glomerular filtration rate in hydrated as well as in nonhydrated rats. The potency and magnitude of the bremazocine induced diuresis were more pronounced in the nonhydrated group of rats. That bremazocine has a central component of action is deduced from the fact that 0.1 microgram of the opioid administered centrally caused a significant increase in urine output; proportionally, larger doses of bremazocine were required to produce the same diuretic effect when the drug was administered parenterally. Bremazocine did not change the total amount of urinary Na+ and K+ as compared to the saline controls; it increased significantly the free water clearance. The bremazocine-induced diuresis was antagonized in a competitive fashion by 10 mg/kg of naloxone giving further support to the notion that the mechanism of action of bremazocine involves activation of kappa-opioid receptors. Bremazocine injected i.v. to nonanesthetized rats increased mean systemic blood pressure in a dose dependent manner; the pressor action of the opiate was blocked and prevented by 1 mg/kg of naloxone. In contrast, i.c.v. administration of bremazocine did not change mean systemic blood pressure but produced a dose-related increase in urine output. To determine whether in addition to a central site bremazocine also activates a renal mechanism, experiments were performed in the isolated perfused rat kidney. Bremazocine (0.15-2.5 microM) caused a dose-dependent diuretic response and a significant rise in perfusion pressure as well as in glomerular filtration rate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550626 TI - Temperature and synaptic efficacy in frog skeletal muscle. AB - 1. Intracellular recording and voltage-clamp techniques were used to measure synaptic efficacy and the safety factor for neuromuscular transmission in frog skeletal muscle. All measurements were made in normal Ringer solution, in the absence of presynaptic or postsynaptic blocking agents. 2. Over a broad temperature range (10-30 degrees C), a small percentage of sartorius fibres (about 6%) could be found which produced only subthreshold end-plate potentials and no action potential in response to single, supramaximal nerve shock. At lower temperatures the proportion of such fibres increased; 42% of the fibres had subthreshold transmission at 5 degrees C, and 59% were subthreshold at 2.5 degrees C. 3. Threshold current, measured by intracellularly injecting short pulses of depolarizing current at end-plate regions, was independent of temperature between 2.5 and 20 degrees C. Thus, the reduced synaptic efficacy observed at low temperatures was not due to decreased electrical excitability of the postsynaptic membrane. 4. The amplitude of evoked end-plate currents (EPCs) decreased with cooling. At temperatures below 10 degrees C, the evoked EPCs at many end-plates were too small to initiate action potentials. The decline in EPC amplitude was due to three factors: a decrease in the amplitude of single quantum currents (MEPCs), an increase in the temporal dispersion of transmitter release, and (below 5 degrees C) a decrease in quantal content. 5. The safety factor for neuromuscular transmission decreased dramatically as temperature was lowered. At 30 degrees C average safety factor was large and positive (540 nA), but at 2.5 degrees C it was negative (-78 nA). 6. The quantal content of evoked transmitter release was independent of temperature change between 5 and 30 degrees C, the average value over this range being 180. However, at temperatures below 5 degrees C, quantal content fell off sharply (average value = 37). 7. The thermal independence of transmitter release may be an important mechanism in allowing poikilothermic animals to maintain physiological function over a wide range of body temperatures. PMID- 2550627 TI - Evidence for the ordered release of rubidium ions occluded within individual protomers of dog kidney Na+,K+-ATPase. AB - 1. When magnesium and orthophosphate are added to Na+,K+-ATPase containing occluded rubidium ions, and suspended in a medium containing free rubidium ions, only 50% of the occluded rubidium is released rapidly. This is because the release of occluded rubidium is ordered, and the replacement (by rubidium ions from the medium) of the first occluded rubidium ions to leave slows the departure of the remaining occluded ions. 2. Since the Na+,K+-ATPase probably exists in the membrane as a structural dimer, the ordered release might represent either the ordered emptying of the two halves of the dimer, or the ordered release of the two rubidium ions thought to be contained in each promoter. 3. The present experiments were designed to decide between these possibilities by examining the behaviour of Na+,K+-ATPase in which about half of the protomers had been randomly inactivated by pre-treatment either with fluorescein isothiocyanate or with alpha chymotrypsin. 4. The results show that the release of rubidium ions from each protomer is ordered. PMID- 2550628 TI - Enhancement of dopamine actions on rat nucleus accumbens neurones in vitro after methamphetamine pre-treatment. AB - 1. Intracellular recordings were made from the nucleus accumbens neurons in brain slices from rats previously treated with saline or methamphetamine. 2. In neurones from both methamphetamine- and saline (control)-treated rats, dopamine (0.1 mM) produced three types of responses: a biphasic response consisting of an initial hyperpolarization followed by a depolarization, a monophasic hyperpolarization and a simple depolarization. 3. Haloperidol (1 microM) reversibly suppressed both responses to dopamine; (-)-sulpiride (1 microM) selectively abolished the depolarization and prolonged the hyperpolarization. Forskolin (10 microM) and dibutyryl adenosine 3',5'-cyclic monophosphate (1 mM) mimicked the hyperpolarization. Both of the latter two substances were more effective in neurones from methamphetamine-treated rats than in neurones from control rats. 4. In slices from methamphetamine-treated rats, the dose-response curve for the dopamine hyperpolarization was shifted to the left of that seen in neurones from control rats by a factor of approximately 100. The dose-response curve for the dopamine depolarization was shifted to the right about 10-fold in neurones from rats treated with methamphetamine. 5. In slices from control rats, dopamine (less than or equal to 0.1 mM) and methamphetamine (less than or equal to 1 microM) had no effect on the EPSPs evoked by focal electrical stimulation of the periaccumbens regions: dopamine (greater than or equal to 10 nM) and methamphetamine (1 microM) markedly depressed the EPSPs in slices from methamphetamine-treated rats. Depolarizations evoked by application of exogenous glutamate were unaffected by dopamine (less than 5 microM). 6. In slices from methamphetamine-treated rats, dopamine (greater than or equal to 10 nM), forskolin (greater than or equal to 1 microM) and dibutyryl adenosine 3',5' cyclic monophosphate (1 mM) depressed Ca2+-dependent spikes as well as the EPSPs. Haloperidol (1 microM) completely reversed the depressions of the EPSPs and Ca2+ dependent spikes by dopamine, while (-)-sulpiride (1 microM) was only partially effective. 7. These results indicate that chronic methamphetamine administration leads to enhancement of the actions of dopamine at D1 receptors located on glutamate and/or aspartate nerve terminals and of the dopamine hyperpolarization of principal neurones, which is also mediated by D1 receptors. PMID- 2550630 TI - Anti-DNA and antilymphocyte antibodies during acute infection with human parvovirus B19. AB - Sera from patients in the acute and recovery stages of parvovirus B19 infection, and from individuals with no detectable antiparvovirus antibody were examined for the presence of anti-DNA and antilymphocyte antibodies. Sixty-eight percent of individuals recently recovered from parvovirus infection had elevated levels of antidouble stranded (ds) and antisingle stranded (ss) DNA antibodies. In addition, a cytotoxic IgM antilymphocyte antibody was detected in more than 88% of these same sera. Serial specimens from volunteers experimentally infected with parvovirus B19 were also tested for these autoantibodies and it was determined that the presence of antilymphocyte IgM was dependent on the stage of infection. The antilymphocyte IgM was occasionally detectable in sera containing rubella specific IgM (11%) or varicella zoster specific IgM (25%). However, in contrast to B19 infection, these antibodies were not cytotoxic. From the results of our study, we propose that parvovirus infection of hematologically normal individuals may be accompanied by a transient, subclinical autoimmune state. PMID- 2550629 TI - Suppression of acute and chronic inflammation by dietary gamma linolenic acid. AB - We examined the effect of diets enriched in gamma linolenic acid (GLA) on acute inflammation induced by monosodium urate crystals, and on subacute and chronic inflammation induced by complete Freund's adjuvant in the rat subcutaneous air pouch and in rats with adjuvant induced arthritis. Diets were enriched (15% fat) with borage seed oil (23% GLA) or safflower oil (less than 1% GLA). Diets enriched with GLA suppressed inflammation markedly in all models, whereas the safflower oil diet did not influence the inflammatory response. The degree of inflammation was quantified by measuring pouch exudate cell concentration, lysosomal enzyme activity, volume, protein concentration and prostaglandin E2 and leukotriene B4 concentrations. In the chronic air pouch model, the pouch lining was thickened, invaded by mononuclear cells and exhibited proliferation of lining cells 14 days after adjuvant injection. The lesion was far less severe and usual pouch lining architecture was maintained in animals given dietary GLA. Livers of rats fed borage seed oil were enriched in GLA and dihomo gamma linolenic acid (DGLA), and the DGLA/arachidonate ratio was increased 5-fold compared with animals fed safflower oil. Enrichment of diet with plant seed oils rich in GLA may provide a way to alter generation of prostaglandins and leukotrienes and to influence acute and chronic inflammatory responses. PMID- 2550631 TI - Calcium pyrophosphate and monosodium urate crystal interactions with neutrophils: effect of crystal size and lipoprotein binding to crystals. AB - "Small" (between 75-98% of crystals less than or equal to 10 microns) and "large" (between 81-93% of crystals greater than 10 microns) size fractions of monosodium urate monohydrate (MSUM) and calcium pyrophosphate dihydrate triclinic (CPPD) crystals were incubated with human neutrophils and crystal induced neutrophil cytolysis monitored by measuring the release of lactate dehydrogenase. "Small" size fractions of MSUM and CPPD gave higher percent lysis values than "large" crystals. The binding of high density lipoproteins (HDL) and low density lipoproteins (LDL) to the crystals was quantitated. HDL and LDL bound in significant amounts to both CPPD and MSUM and strongly inhibited CPPD and MSUM induced neutrophil cytolysis. We propose that HDL and LDL bound to MSUM and CPPD may play important roles in the regulation of gouty inflammation. PMID- 2550632 TI - A case of breast carcinoma in association with in vitro fertilization. PMID- 2550633 TI - ACE inhibition--the next decade. Focus on lisinopril. Proceedings of an international symposium. Cannes, France, 4-6 November 1988. PMID- 2550635 TI - The clinical pharmacology of lisinopril. AB - Structurally, lisinopril differs from captopril in that it does not contain a sulphydryl group and it differs from enalapril and related compounds in that it is not an ester prodrug. Published data on the clinical pharmacology of lisinopril are reviewed and data from new studies are presented. A radioimmunoassay has been used to study the clinical pharmacokinetics of lisinopril and 14C-lisinopril has been used in metabolism studies in man. Following oral administration, lisinopril is absorbed slowly but the absorbed drug is immediately available without any requirement for biotransformation by the liver. The plasma half-life controlling accumulation during chronic administration is 12-13 h and the absorbed drug is eliminated via glomerular filtration. These properties are consistent with once-daily dosing and uncomplicated clinical use in the treatment of hypertension and congestive heart failure. PMID- 2550634 TI - Assessment of the antihypertensive effect of lisinopril using 24-hour ambulatory monitoring. AB - The effect of lisinopril 20 mg given once daily was assessed in 25 patients with mild to moderate essential hypertension using both ambulatory monitoring and cuff measurements of blood pressure (BP) made in the clinic. The effect of 4 weeks of lisinopril treatment on BP was compared with the BP recorded after 2 weeks on placebo. Lisinopril treatment reduced supine and standing cuff clinic measurements of BP 24 h after dosing by (systolic/diastolic) 21.1/11.8 and 16.7/10.1 mmHg, respectively. Ambulatory measurements indicated that the antihypertensive effect of lisinopril was sustained throughout the 24 h without any significant effect on heart rate and that lisinopril did not affect the diurnal rhythm of BP changes. Side effects reported during the 4 weeks of lisinopril treatment were mild and did not necessitate discontinuation of treatment. Lisinopril appears to be an effective and well-tolerated antihypertensive agent when given in a single daily dose. PMID- 2550636 TI - Comparative pharmacokinetics and pharmacodynamics of lisinopril and enalapril, alone and in combination with propranolol. AB - In an open, crossover study, the pharmacokinetic and pharmacodynamic profiles of lisinopril and enalapril, administered alone and in combination with propranolol, were evaluated in 12 volunteers. The maximum serum concentration (Cmax) of lisinopril and time to reach maximum concentration (Tmax) were 64 +/- 16 ng/ml and 7.5 +/- 1.5 h, respectively. The area under the serum curve (AUC) was 916 +/- 239 h. ng/ml. The Cmax of enalaprilat (89 +/- 34 ng/ml) was greater than that of lisinopril whilst Tmax was shorter (4.3 +/- 1.7 h) and AUC smaller (718 +/- 17 h.ng/ml) (P less than 0.01). Renal clearance of drug 48 h post-dosing showed that enalaprilat (164 +/- 38 ml/min) was cleared from plasma significantly more rapidly than lisinopril (82 +/- 16 ml/min) (P less than 0.001). Mean supine blood pressure decreased significantly with all treatments, as did heart rate. No significant changes were observed in either the serum concentrations or the urinary outputs of these ACE inhibitors following combination with propranolol, apart from a greater variability of Cmax after addition of propranolol to enalapril compared with lisinopril in combination. PMID- 2550637 TI - Comparative pharmacokinetics of enalapril and lisinopril, alone and with hydralazine. AB - A single-dose, single-blind, crossover study of vasodilator/angiotensin converting enzyme (ACE) inhibitor interactions was carried out in 16 volunteers. Enalapril 20 mg and lisinopril 20 mg were administered either alone or in combination with hydralazine 25 mg. Co-administration of hydralazine significantly increased the area under the plasma concentration time curve (AUC) of lisinopril (AUC0-48 h 766.8 +/- 66.3 ng.h/ml (lisinopril) vs 1022.3 +/- 115.3 ng.h/ml (lisinopril + hydralazine)). This did not occur with enalaprilat (AUC 710.1 +/- 51.2 ng.h/ml (enalapril) vs 681.9 +/- 44.9 ng.h/ml (enalapril + hydralazine); mean +/- SEM). Urinary recovery of lisinopril showed a similar trend, but group differences did not achieve statistical significance despite comparable confidence intervals. Although hydralazine had no effect on the bioavailability of enalapril, significantly increased bioavailability was observed with lisinopril. This type of drug interaction is rare. The underlying mechanism is unclear, but may relate to increased absorption of lisinopril. PMID- 2550638 TI - Pharmacokinetics of enalapril and lisinopril in subjects with normal and impaired hepatic function. AB - The pharmacokinetic and pharmacodynamic profiles of two angiotensin-converting enzyme (ACE) inhibitors, enalapril (a prodrug) and lisinopril (directly acting), were compared in eight patients with hepatic cirrhosis and 10 healthy controls. The pharmacokinetics of both drugs were affected in patients with hepatic cirrhosis. The percentage urinary recovery of the parent (inactive) drug enalapril was higher in patients with cirrhosis than controls. Serum concentrations of both drugs showed considerable variation in cirrhotic patients, but the variance ratio between patients with cirrhosis and controls was greater for enalapril than lisinopril. Peak serum concentrations of both ACE inhibitors were higher in patients with cirrhosis than in controls, which may be due to increased drug absorption. The time to peak drug concentration was longer for lisinopril than enalapril. PMID- 2550639 TI - Safety and efficacy of lisinopril in elderly patients with mild to moderate hypertension. AB - The safety and efficacy of once-daily lisinopril was assessed in an 8-week open label study of 24 elderly patients with mild to moderate hypertension. Following withdrawal of all previous antihypertensive treatment, including diuretics, lisinopril treatment was started at a dose of 5 mg, increasing to a maximum of 40 mg once daily as required. Treatment provided effective 24-h BP control in all patients, most of whom required a daily dose of 20-40 mg, and cardiothoracic index decreased significantly, indicating a favourable effect on left ventricular volumes. There were no significant ECG alterations and no major side effects occurred. It is concluded that lisinopril 5 mg is a safe starting dose for elderly patients who are not on diuretics, and that once-daily lisinopril monotherapy reduces BP safety without affecting heart rate. PMID- 2550640 TI - Lisinopril in the treatment of hypertension. AB - Established essential hypertension is characterised haemodynamically by a normal cardiac output and elevated total peripheral resistance. As hypertensive cardiovascular disease progresses, and the patient grows older, cardiac output falls and total peripheral resistance is further elevated. The activity of the renin-angiotensin-aldosterone (RAA) system declines throughout life and reaches its lowest levels in the elderly, unless there is congestive heart failure. In long-standing hypertension, target organ disease such as left ventricular hypertrophy, nephrosclerosis and cerebrovascular damage is commonly observed. Rational antihypertensive therapy should therefore aim to lower total peripheral resistance, spare cardiac output, and maintain or improve blood flow to target organs. ACE inhibitors lower arterial pressure by decreasing total peripheral resistance, they maintain or improve cardiac contractility, promote regression of left ventricular hypertrophy, and increase renal blood flow. Lisinopril is a novel ACE inhibitor that does not contain a sulphydryl group. It is not a prodrug and thus does not require bioactivation by the liver. Lisinopril has a long duration of action, allowing it to be used as a single daily dose in the treatment of hypertension. Preliminary studies from our laboratory indicate that lisinopril reduces cardiac output and preload to the left ventricle. Lisinopril also reduces left ventricular hypertrophy and lowers renal vascular resistance, thereby increasing renal blood flow. In patients with mild to moderate hypertension, lisinopril is more effective than hydrochlorothiazide in reducing both systolic and diastolic blood pressure, and is at least as effective as atenolol or metoprolol in reducing diastolic blood pressure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550641 TI - Clinical experience with lisinopril. Observations on safety and tolerability. AB - The safety and tolerability of lisinopril (1.25-160 mg daily) were assessed in 3,270 patients (2,688 hypertensives and 582 patients with congestive heart failure (CHF] and 280 healthy subjects. The duration of therapy ranged from a single dose to 43 months; 438 patients received lisinopril for at least 12 months (mean 20 months). In the hypertensive population, the most frequent adverse events reported were headache, dizziness, cough, nausea, diarrhoea and fatigue, although not all of these events were thought to be related to lisinopril; 6.1% discontinued lisinopril due to adverse clinical events, most commonly cough and nausea. Twelve hypertensive patients died (0.45%), but most of these were not receiving lisinopril at the time of death and none was considered to be drug related. In CHF patients, the most frequently reported adverse events were dizziness, dyspnoea, diarrhoea, hypotension and fatigue. Again, not all of these reports were considered to be drug-related. Therapy was withdrawn in 9.6% of patients--hypotension, dizziness, diarrhoea and rash being the most frequent reasons. Fifty-three CHF patients died (9.1%) and in three cases death was considered to be related to lisinopril therapy. Hypotension, orthostatic effects or dizziness following the initial dose of lisinopril occurred infrequently (in 1.3% of the hypertensive group, including those receiving hydrochlorothiazide, and in 4.8% of CHF patients). Changes in laboratory parameters were generally minor and seldom resulted in discontinuation of therapy. Long-term treatment of hypertension and CHF with lisinopril for at least 3 years confirms that the drug is well tolerated. Overall, the side-effect profile is very similar to that of other ACE inhibitors with regard to class-specific effects. However, taste disturbance was rarely observed. PMID- 2550642 TI - ACE inhibitors in mild to moderate hypertension: comparison of lisinopril and captopril administered once daily. French Cooperative Study Group. AB - A double-blind, parallel-group, multicentre study was carried out to compare the effects of once-daily treatment with lisinopril 20 mg and captopril 50 mg in 304 patients with mild to moderate hypertension. Following a 2-week placebo run-in period, patients with a lying DBP between 95 and 115 mmHg were randomised to either active treatment for 10 weeks. BP measurements were made approximately 24 h after the previous dose of either drug. After 6 weeks, hydrochlorothiazide 25 mg once daily was added for patients with a lying DBP greater than or equal to 95 mmHg and a final assessment was made in all patients after a further 4 weeks of treatment. After 6 weeks of monotherapy, lisinopril had reduced lying and standing BP by an average of 18/14 mmHg and 19/12 mmHg, respectively, compared with 15/12 mmHg and 14/11 mmHg respectively for captopril. The larger fall in SBP with lisinopril was statistically significant (lying SBP, P = 0.01; standing SBP, P = 0.0001). In addition, a significantly larger proportion of patients achieved BP control (DBP less than 95 mmHg) with lisinopril than with captopril (79% versus 67%; P = 0.02). Neither drug significantly altered heart rate. The addition of hydrochlorothiazide in some patients produced a further, small reduction in mean BP in both groups although, 10 weeks after randomisation, the proportion of patients in whom BP control had been achieved with lisinopril was still significantly greater than that achieved with captopril (90% versus 79%; P = 0.02). Both drugs were well tolerated and no serious adverse events occurred during the study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550643 TI - Lisinopril versus slow-release nifedipine in the treatment of mild to moderate hypertension: a multicentre study. The Cooperative Study Group. AB - The antihypertensive effects of lisinopril 20 mg once daily and slow-release nifedipine 20 mg twice daily were compared in a double-blind, parallel group, 10 week study involving 274 patients with mild to moderate hypertension. During the first 6 weeks of treatment, slow-release nifedipine and lisinopril produced similar reductions in lying and standing blood pressure (BP), except for lying systolic BP (SBP) which was reduced to a greater extent by lisinopril. After 6 weeks of double-blind treatment, hydrochlorothiazide 25 mg once daily was added if BP remained uncontrolled (lying DBP greater than or equal to 95 mmHg); a significantly greater proportion of patients in the nifedipine group than in the lisinopril group required additional diuretic treatment (29% versus 14%, respectively; P = 0.005). Moreover, after a further 4 weeks of treatment BP was adequately controlled (lying DBP less than 95 mmHg) in significantly more lisinopril-treated patients than in the nifedipine group (91.4% versus 78.3%, respectively; P = 0.006). Lisinopril was better tolerated than slow-release nifedipine. The frequency of drug-related events was significantly lower (threefold) for lisinopril than for nifedipine (P = 0.001) and the number of withdrawals from treatment with nifedipine was more than three times that in the lisinopril treatment group (P = 0.009). Lisinopril appears to provide an effective once-daily antihypertensive treatment which is at least as effective as, and better tolerated than, slow-release nifedipine. PMID- 2550644 TI - A comparison of single doses of lisinopril and enalapril in hypertension. AB - Sixteen patients with mild to moderate hypertension were studied in a double blind crossover comparison of single oral doses of lisinopril 10 mg and enalapril 10 mg. Both drugs caused a marked fall in blood pressure (BP) with a clinically useful effect persisting for 24 h postdose and with no significant difference between treatments in this respect. The time to minimum systolic BP was 2 h (95% confidence limits 0-19 h) longer for lisinopril. The fall in ACE activity at 24 h was 18.5 (8.2-28.8) U/l greater for lisinopril. We conclude that, within the limits of this small study, lisinopril appears to be as effective as enalapril in lowering BP and that it may have a slower onset of action that could be clinically valuable. PMID- 2550645 TI - Angiotensin-converting enzyme inhibition and compliance of the carotid artery in normotensive and hypertensive rats. AB - To investigate the effects of local application of an angiotensin-converting enzyme inhibitor on the mechanical properties of the arterial wall, the volume/pressure relationship (from 25 to 175 mmHg) in the in vivo isolated carotid artery was measured in normotensive (WKY) and spontaneously hypertensive rats (SHR). The compliance of the carotid artery (CC) was calculated, for each level of pressure, as the slope of the the volume/pressure curve. The carotid artery was less compliant in the SHR strain than in normotensive rats. This rigidity was partly related to an increased tone of the arterial smooth muscle in SHR compared with the WKY. Local incubation with lisinopril (2.3 micrograms/ml) produced a significant increase in the compliance of the carotid artery in both normotensive and hypertensive rats; CC was increased by 23% in WKY rats and by 14% in SHR. Since the goal of antihypertensive treatment may be not only to reduce blood pressure (BP) but also to improve the mechanical properties of the arterial system, the direct effect of lisinopril on the arterial wall may have a major beneficial role. PMID- 2550646 TI - Mechanisms for improved survival in heart failure. AB - The two major causes of death in congestive heart failure (CHF) are progressive heart failure (approximately 60% of cases) and sudden death (30%). Sudden death in CHF is caused primarily by malignant ventricular arrhythmias. The underlying mechanism has yet to be established, but myocardial metabolic factors are probably involved. Although there is a clear association between complex ventricular arrhythmias and left ventricular function, it has not been shown convincingly that antiarrhythmic agents can reduce sudden death in CHF. Progressive deterioration of myocardial function is associated with altered myocardial energy production. Notably, the physiological effects of neuroendocrine activation in chronic CHF may be deleterious to myocardial function. The CONSENSUS study was carried out to evaluate the association between neuroendocrine activation and deterioration of myocardial function using ACE inhibitors. A marked reduction in mortality rate occurred in the enalapril treated group, where the one-year mortality was reduced by 31%. The reduction in mortality was solely among patients with progressive CHF (a reduction of 50%); there was no difference in the incidence of sudden death. Analysis of blood samples drawn at baseline in the placebo group showed a significant positive correlation between mortality and plasma angiotensin II (P less than 0.05), aldosterone (P = 0.003), noradrenaline (P less than 0.001), adrenal levels (P less than 0.001), and atrial natriuretic peptide (P = 0.003). This was not observed in enalapril-treated patients. The significant reduction in mortality in the enalapril group was found consistently among patients with baseline hormone levels above the median value. In CHF, the underlying disease may induce serious arrhythmias and/or progressive failure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550647 TI - Lisinopril in the treatment of congestive heart failure. AB - In a series of studies, lisinopril, a new, once-daily, non-sulphydryl-containing ACE inhibitor, has been evaluated for the treatment of congestive heart failure (CHF). Lisinopril significantly improved exercise duration, left ventricular ejection fraction (LVEF) and the clinical signs and symptoms of CHF when compared with placebo in a 12-week prospective, randomised, double-blind trial involving 130 patients (NYHA Class II-IV) who were also receiving digitalis and diuretics. In another study involving 10 patients with CHF, lisinopril was found to increase maximal oxygen uptake. Lisinopril has also been compared with captopril in a 12 week randomised, double-blind, parallel trial in 189 patients with CHF (Class II IV) on digoxin and diuretics. Both lisinopril and captopril increased treadmill exercise times. However, lisinopril was equally effective in patients with renal impairment whereas captopril was not. Also, lisinopril, but not captopril, improved LVEF. In all studies, lisinopril was well tolerated in a manner similar to captopril. Lisinopril produces more frequent increases in serum creatinine than captopril, but these changes are rarely of significant consequence. These results confirm those of previous studies, from which it is concluded that lisinopril is an important addition to the ACE inhibitor class for the treatment of CHF. PMID- 2550648 TI - Scanning electronmicroscopy of Staphylococcus aureus and Enterococcus faecalis exposed to daptomycin. AB - The novel lipopeptide antibiotic, daptomycin, at a concentration of 8 mg/L, caused gross morphological changes in both a methicillin-sensitive and a methicillin-resistant strain of Staphylococcus aureus and in a strain of Enterococcus faecalis. The earliest (after 1 h) surface lesion observed was the appearance of boss-like processes randomly distributed on the cell surface. Later, grossly deformed bacteria were seen and in two of the three bacteria prolonged exposure led to degeneration of the cells into an amorphous syncytial mass. Omission of calcium (which is known to potentiate the activity of daptomycin) from the culture medium did not affect the morphological response to an inhibitory concentration of the antibiotic. PMID- 2550649 TI - Isoforms of Na,K-ATPase in Artemia saline: I. Detection by FITC binding and time course. AB - Partially purified Na,K-ATPase from whole nauplii at various stages of development, analyzed by SDS-PAGE, reveals a polydisperse beta and two alpha subunits (denoted alpha 1 and alpha 2). In the absence of Ca2+, ATP-inhibitable fluorescein isothiocyanate (FITC) labeling is restricted to the alpha subunit of this enzyme, even in crude naupliar homogenates. The intensity of the alpha specific fluorescent signal (i.e., the sum of the yield from both alpha isoforms) is proportional to Na,K-ATPase activity during development. FITC-labeled subunits were detected at 8 hr of development prior to the detection of measurable Na,K ATPase activity. The alpha 2/alpha 1 ratio changed from an initial value of 1.25 to a peak of 1.75 at 32 hr of development, then reverted to a ratio of 1.25 by 42 hr, and remained constant thereafter. Pulse chase studies with 35S-methionine indicated that the developmental increase in enzyme activity is coincident with amino acid incorporation into the alpha subunits, implying that enzyme synthesis is active during enzyme accumulation. PMID- 2550650 TI - Isoforms of Na,K-ATPase in Artemia salina: II. Tissue distribution and kinetic characterization. AB - To characterize the molecular properties conveyed by the isoforms of the alpha subunit of Na,K-ATPase, the two major transepithelial transporting organs in the brine shrimp (Artemia salina), the salt glands and intestines, were isolated in pure form. The alpha isoforms were quantified by ATP-sensitive fluorescein isothiocyanate (FITC) labeling. The salt gland enzyme exhibits only the alpha 1 isoform, whereas the intestinal enzyme exhibits both the alpha 1 and the alpha 2 isoforms. After 32 hours of development, Na,K-ATPase activity [in mumol Pi/mg protein/hr (1 mu)] in whole homogenates was 32 +/- 6 in the salt glands and 12 +/ 3 in the intestinal preparations (mean +/- SEM). The apparent half-maximal activation constants (K1/2) of the salt gland enzyme as compared to the intestinal enzyme were 3.7 +/- 0.6 mM vs. 23.5 +/- 4 mM (P less than 0.01) for Na+, 16.6 +/- 2.2 mM vs. 8.29 +/- 1.5 mM for K+ (P less than 0.01), and 0.87 +/- 0.8 mM vs. 0.79 +/- 1.1 mM for ATP (NS). The apparent Ki's for ouabain inhibition were 1.1 x 10(-4) M vs. 2 x 10(-5) M, respectively. Treatment of whole homogenates with deoxycholic acid (DOC) produced a maximal Na,K-ATPase activation of 46% in the salt gland as compared to 23% in the intestinal enzyme. Similar differences were found with sodium dodecyl sulfate (SDS). The two distinct forms of Na,K-ATPase isolated from the brine shrimp differed markedly in three kinetic parameters as well as in detergent sensitivity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550651 TI - Role of calcium and calmodulin in the regulation of the rabbit ileal brush-border membrane Na+/H+ antiporter. AB - In rabbit ileum, Ca2+/calmodulin (CaM) appears to be involved in physiologically inhibiting the linked NaCl absorptive process, since inhibitors of Ca2+/CaM stimulate linked Na+ and Cl- absorption. The role of Ca2+/CaM-dependent phosphorylation in regulation of the brush-border Na+/H+ antiporter, which is believed to be part of the neutral linked NaCl absorptive process, was studied using purified brush-border membrane vesicles, which contain both the Na+/H+ antiporter and Ca2+/CaM-dependent protein kinase(s) and its phosphorprotein substrates. Rabbit ileal villus cell brush-border membrane vesicles were prepared by Mg precipitation and depleted of ATP. Using a freezethaw technique, the ATP depleted vesicles were loaded with Ca2+, CaM, ATP and an ATP-regenerating system consisting of creatine kinase and creatine phosphate. The combination of Ca2+/CaM and ATP inhibited Na+/H+ exchange by 45 +/- 13%. This effect was specific since Ca2+/CaM and ATP did not alter diffusive Na+ uptake, Na+-dependent glucose entry, or Na+ or glucose equilibrium volumes. The inhibition of the Na+/H+ exchanger by Ca2+/CaM/ATP was due to an effect on the Vmax and not on the Km for Na+. In the presence of CaM and ATP, Ca2+ caused a concentration-dependent inhibition of Na+ uptake, with an effect 50% of maximum occurring at 120 nM. This Ca2+ concentration dependence was similar to the Ca2+ concentration dependence of Ca2+/CaM-dependent phosphorylation of specific proteins in the vesicles. The Ca2+/CaM/ATP-inhibition of Na+/H+ exchange was reversed by W13, a Ca2+/CaM antagonist, but not by a hydrophobic control, W12, or by H-7, a protein kinase C antagonist. We conclude that Ca2+, acting through CaM, regulates ileal brush border Na+/H+ exchange, and that this may be involved in the regulation of neutral linked NaCl absorption. PMID- 2550652 TI - Cross-linking of cardiac gap junction connexons by thiol/disulfide exchanges. AB - SDS-polyacrylamide gel electrophoresis and immunoblotting were used to investigate inter- and intramolecular disulfide bonds to connexin 43 (the cardiac gap junctional protein) in isolated rat heart gap junctions and in whole heart fractions. In gap junctions isolated in the absence of alkylating agent, connexin 43 molecules are cross-linked by disulfide bonds. The use of iodoacetamide (100 mM) for the first steps of isolation procedure prevents the formation of these artifactual linkages. Investigation of connexin 43 in whole heart fractions by means of antibodies confirms the results obtained with isolated gap junctions; that is, connexin 43 molecules are not interconnected with disulfide bridges. In whole heart fractions treated with alkylating agents, a 38 kD protein, immunologically related to connexin 43, and containing intramolecular disulfide bonds is detected. It is hypothesized that this protein might be a folded form of connexin 43, a precursory form of the molecules embedded in the gap junctions. PMID- 2550653 TI - Subcellular distribution of high-affinity type IV cyclic AMP phosphodiesterase activities in rabbit ventricular myocardium: relations to post-natal maturation. AB - Cytosolic and particulate Type IV (high-affinity) cAMP phosphodiesterase (PDE) activities were isolated from the ventricular myocardium of newborn (NB; 24 to 48 h), immature (IM; 14 to 16 days) and adult (AD; 6 to 8 months) rabbits. Cytosolic activity from each age group was resolved into three distinct peaks of activity by DEAE cellulose anion exchange chromatography. Type IV PDE activity was identified as a predominant activity in the cytosolic peak III activity in all three age groups when measured with 0.25 microM cAMP as substrate. A particulate Type IV PDE activity was associated with the sarcoplasmic reticulum (SR) fractions in each age group. No significant age-related changes in the affinity of the particulate enzyme for cAMP (apparent Km = 0.3 to 0.5 microM) were evident, but the Vmax for this SR-associated activity increased from 553 +/- 7 pmol/min/mg in the NB to 725 +/- 9 pmol/min/mg in the IM and 2450 +/- 33 pmol/min/mg in the AD. In each age group, milrinone, imazodan, piroximone and indolidan were more potent inhibitors of the SR-associated activity as compared with the cytosolic peak III activity. In contrast, RO 20-1724 and rolipram were relatively more selective inhibitors of the cytosolic peak III activity. Age related differences in the sensitivity of type IV PDE to inhibition was dependent upon the selectivity of the inhibitor and the subcellular enzymic distribution. Cytosolic peak III PDE activity was further resolved by gel filtration chromatography into two peaks. Hydrolysis of cAMP by the higher molecular weight peak was inhibitable by cGMP (IC50 = 0.25 +/- 0.07 microM in NB and 0.07 +/- 0.01 microM in AD) whereas the lower molecular weight peak activity was relatively insensitive to inhibition by cGMP (IC50 greater than 100 microM). The lower molecular weight peak constituted a relatively greater proportion of the total peak III activity in the NB as compared to the AD. Analysis of the kinetics of cGMP inhibition of high-affinity cAMP hydrolysis was consistent with the presence of a greater number of high-affinity (presumably drug-sensitive) binding sites in the SR-associated activity as compared to the cytosolic peak III activity in both NB and AD. These results support the hypothesis that the cGMP-inhibitable Type IV PDE activity may be the primary site of action for certain newer cardiotonic drugs. Differences in drug action in young versus adult myocardium may be related to the selectivity of the cardiotonic drugs for this specific isozyme and its lower specific activity during the early stages of maturation. PMID- 2550654 TI - Autonomic receptor interactions in isolated cardiac myocytes from hypertensive rats. AB - Isolated ventricular myocytes from adult (16 to 20 weeks) spontaneously hypertensive (SHR) and normotensive (WKY) rats were utilized to examine adrenergic and cholinergic receptor expression and interaction. Binding assays were performed using quinuclidinyl benzilate (QNB) and iodocyanopindolol (ICYP) for cholinergic and beta-adrenergic receptors, respectively. In addition, cAMP was measured as an index of adrenergic-cholinergic control of adenylate cyclase. Data from radioligand binding experiments indicated that muscarinic cholinergic receptors were depressed (22%) in SHR myocytes, while beta-adrenergic receptor density was comparable to that of WKY myocytes. Heterologous receptor modulation in isolated myocytes as assessed by displacement analysis with and without guanosine 5'-triphosphate (GTP), showed that carbachol displacement of QNB was shifted five fold to the right in the presence of GTP and that the beta adrenergic agonist isoproterenol did not prevent the GTP-mediated binding alteration. In contrast, carbachol modulated the GTP-shift of ICYP displacement by isoproterenol and these effects were comparable in both WKY and SHR myocytes. Furthermore, the ability of carbachol to blunt the stimulation of adenylate cyclase by isoproterenol was also comparable in myocytes isolated from adult SHR and control animals. Thus, the observed decrement in muscarinic cholinergic receptor expression did not alter adrenergic-cholinergic interactions as assessed by displacement assays using guanine nucleotides, or the control of cAMP levels. In addition, isolated myocytes provide a useful system for analyzing receptor expression and regulation and how these parameters may be altered in the hypertensive heart. PMID- 2550655 TI - Metabolism of dioctanoylglycerol by isolated cardiac myocytes. AB - Diacylglycerol (DG) metabolism by intact cardiac myocytes isolated from adult rat hearts and by broken cell preparations from myocytes was investigated in experiments with [3H]dioctanoylglycerol (diC8), a cell-permeable diacylglycerol analog. In a low-speed supernatant fraction, the Km and Vmax for DG kinase activity (formation of phosphatidic acid) was 22 microM and 110 nmol/h/mg, respectively, whereas the Km and Vmax for DG lipase activity (formation of monoacylglycerol) was 80 microM and 1000 nmol/h/mg. At a substrate concentration of 80 microM diC8, lipase activity was 7-fold greater than kinase activity. The majority of DG kinase activity was recovered in the soluble subcellular fraction; DG lipase activity was localized in a microsomal fraction. When [3H]diC8 was incubated with intact cardiac myocytes, 10-fold more radioactivity was incorporated into the products of the lipase pathway (monoacylglycerol and free glycerol) as compared to incorporation into the total phospholipid fraction which contained phosphatidic acid. This predominance of metabolism by hydrolysis through the lipase pathway was observed consistently when the incubation time, content of cardiac myocytes and concentration of exogenous diC8 was varied. Therefore, results from both in vitro determinations of enzyme activities in broken cell preparations and flux studies with intact cells have indicated that the lipase pathway is the principal enzymatic mechanism for the metabolism of diC8 in cardiac myocytes. PMID- 2550657 TI - Variation in salmonid mitochondrial DNA: evolutionary constraints and mechanisms of substitution. AB - Sequence comparisons were made from 2214 bp of mitochondrial DNA cloned from six Pacific salmonid species. These sequences include the genes for ATPase subunit 6, cytochrome oxidase subunit 3, NADH dehydrogenase subunit 3, NADH dehydrogenase subunit 4L, tRNA(GLY), and tRNA(ARG). Variation is found at 338 silent and 12 nonsilent positions of protein coding genes and 10 positions in the two tRNA sequences. A single 3-bp length difference was also detected. In all pairwise comparisons the sequence divergence observed in the fragment was higher than that previously predicted by restriction enzyme analysis of the entire molecule. The inferred evolutionary relationship of these species is consistent between methods. The distribution of silent variation shows a complex pattern with greatly reduced variation at the junctions of genes. The variation in the tRNA sequences is concentrated in the DHU loop. The close relationship of these species and extensive sequence analyzed allows for an analysis of the spectrum of substitutions that includes the frequencies of all 12 possible substitutions. The observed spectrum of substitutions is related to potential pathways of spontaneous substitution. The salmonid sequences show an extremely high ratio of silent to replacement substitutions. In addition the amino acid sequences of the four proteins coded in this fragment show a consistently high level of identity with the Xenopus sequences. Taken together these data are consistent with a slower rate of amino acid substitution among the cold-blooded vertebrates when compared to mammals. PMID- 2550656 TI - The muridae glyceraldehyde-3-phosphate dehydrogenase family. AB - Although only one gene is known to be functional, numerous glyceraldehyde-3 phosphate dehydrogenase (GAPDH) related sequences are scattered throughout Mus musculus and Rattus rattus genomes. In this report we show that: (1) GAPDH pseudogenes are repeated to comparable extents, at least 400 copies, in 12 other Muridae species; (2) the complete, or nearly so, sequence of GAPDH messenger RNA is amplified, and a high proportion, if not all of these copies, are intronless; (3) GAPDH pseudogenes are preferentially located in heavily methylated and DNAse I-insensitive regions of chromatin; and (4) the presence of atypical GAPDH related mRNAs in different cellular contexts raises the possibility that more than one GAPDH gene is transcribed. PMID- 2550658 TI - Targeted toxin therapy for the treatment of cancer. AB - Protein toxins such as Pseudomonas exotoxin, diphtheria toxin, and ricin may be useful in cancer therapy because they are among the most potent cell-killing agents. One molecule of a toxin delivered to the cytoplasm of a cancer cell will be lethal for that cell. However, to be therapeutically useful, these toxins need to be targeted to specific sites on the surface of cancer cells, then be internalized and ultimately reach the cell cytoplasm. This process is accomplished by eliminating binding to toxin receptors and redirecting the cell killing activity of the toxin to receptors or antigens present on cancer cells. Typically, toxins are conjugated to cell-binding proteins such as monoclonal antibodies or growth factors. These conjugates bind and kill cancer cells selectively while normal cells, which don't bind the conjugates, are spared. Because the genes for many protein toxins have been cloned, it is possible to make genetic modifications to their structure. By deleting the DNA that codes for the toxin binding region and replacing it with various complementary DNA encoding other cell-binding proteins, it has been possible to make chimeric toxins that kill cells on the basis of the newly acquired binding activity. The ability to make these chimeras may be useful in designing future toxin-based anticancer therapies. PMID- 2550659 TI - Primary cancer of the lung in women. AB - Analyses of the St. Francis Medical Center Cancer Registry for a 30-year period corroborated the publicized information of a gradual increase in the incidence of lung cancer in women and a recent decrease in the incidence among men. Our objectives were to highlight the biological and clinical features of lung cancer in women with the adoption and utilization of primary and secondary preventive measures. PMID- 2550661 TI - Loss of Marek's disease virus tumorigenicity is associated with truncation of RNAs transcribed within BamHI-H. AB - The attenuation of Marek's disease virus (MDV) is associated with loss of pathogenicity and tumorigenicity. Previous studies have demonstrated a strong correlation between attenuation and amplification of a specific sequence located within the MDV terminal and internal repeats. We recently reported that the regions containing the amplified sequences, the BamHI D and H fragments, were transcriptionally active. However, differential transcription activity was observed to exist between attenuated and pathogenic MDV strains. Specifically, a major transcript of 1.8 kilobases was found to be produced by pathogenic MDV and not by attenuated MDV. We now report that the disappearance of this transcript is concomitant with the production of a 0.4-kilobase RNA, an RNA resulting from the truncation of the tumorigenicity-related transcript. PMID- 2550660 TI - Epstein-Barr virus (EBV)-containing nasopharyngeal carcinoma cells express the B cell activation antigen blast2/CD23 and low levels of the EBV receptor CR2. AB - Anaplastic nasopharyngeal carcinoma (NPC) cells invariably harbor the Epstein Barr virus (EBV) genome, an association that is unique among human virus associated cancers. Although EBV is able to replicate in epithelial cells, results with expression of the EBV receptor (complement receptor type 2 [CR2]; also called CD21) in normal and malignant epithelial cells are conflicting. We grew five different EBV-associated NPC tumors in nude mice, and by using a sensitive transcriptional assay, we detected a very weak transcription signal of the EBV receptor CR2 gene in these cells. This suggests that low levels of EBV receptor may be expressed by malignant epithelial nasopharyngeal cells. The gene coding for Blast2/CD23, a B-cell activation molecule induced by EBV, was transcribed in three of the transplanted NPC tumors. The soluble form of the Blast2/CD23 protein was also detected in medium taken from short-term cultures of the same NPC cell lines. In contrast to the lymphoid system, in which Blast2/CD23 expression is associated with EBV nuclear antigen (EBNA2) expression, no EBNA2 protein could be detected in these NPC epithelial cells. Our study represents the first demonstration of Blast2/CD23 expression in epithelial cells. As the soluble form of the Blast2/CD23 protein possesses growth factor activity associated with EBV-induced B-cell immortalization, these results suggest a possible role for this molecule in the pathogenesis of NPC. PMID- 2550662 TI - Trojan horse lymphocytes: a vesicular stomatitis virus-specific T-cell clone lyses target cells by carrying virus. AB - We have isolated a vesicular stomatitis virus (VSV)-specific CD4+ CD8- murine T cell clone. This clone proliferates only in response to VSV and lyses infected tumor cells bearing class II major histocompatibility antigens in short-term chromium release assays. In addition, the cell has VSV antigens on its surface and is capable of killing uninfected tumor cells without major histocompatibility antigen restriction in a 2-day assay. This latter cytolytic activity is eliminated by anti-VSV antibody, indicating that its lytic activity is provided by the virus. [35S]methionine labeling and immunoprecipitation experiments demonstrated that viral protein translation is initiated after incubation of the clone with a tumor target cell, defining this as the mechanism of its cytolytic activity. PMID- 2550663 TI - Viral transcription is necessary and sufficient for vesicular stomatitis virus to inhibit maturation of small nuclear ribonucleoproteins. AB - Infection of baby hamster kidney cells with vesicular stomatitis virus (VSV) results in the accumulation of immature U1 and U2 small nuclear ribonucleoproteins (snRNPs) that contain precursor U RNAs and at least some of the proteins specific for U1 and U2 snRNAs but lack the Sm complex of proteins that is common to these U snRNAs. The VSV function required for this effect is not known, but direct inhibition of cellular transcription did not alter the maturation of U1 and U2 snRNPs. On the other hand, viral transcription but not viral translation was required to inhibit U1 and U2 snRNP maturation. Temperature shift experiments with the mutant G114 showed that ongoing viral transcription was necessary, but that viral mRNA was not required for this inhibition. Furthermore, the VSV function involved in the inhibition of maturation of U1 and U2 snRNPs had a small UV target size of approximately 10 to 20 nucleotides. We demonstrate that temperature-sensitive mutants of VSV can be used as a tool to initiate the assembly of snRNPs in infected cells. These results are compatible with the suggestion that perturbation of snRNP metabolism by VSV precedes and is distinct from the effect of VSV on cellular RNA synthesis, although VSV leader RNA may be involved in both these functions. PMID- 2550665 TI - Expression in recombinant vaccinia virus of the equine herpesvirus 1 gene encoding glycoprotein gp13 and protection of immunized animals. AB - The equine herpesvirus 1 (EHV-1) gene encoding glycoprotein 13 (gp13) was cloned into the hemagglutinin (HA) locus of vaccinia virus (Copenhagen strain). Expression of the gp13 gene was driven by the early/late vaccinia virus H6 promoter. Metabolically radiolabeled polypeptides of approximately 47 and 44 kilodaltons and 90 kilodaltons (glycosylated form) were precipitated with both polyclonal and gp13-specific monoclonal antibodies. Presentation of gp13 on the cytoplasmic membrane of cells infected with the recombinant gp13 vaccinia virus was demonstrated by immunofluorescence of unfixed cells. Inoculation of the recombinant gp13 vaccinia virus into guinea pigs induced neutralizing antibodies to both EHV-1 and vaccinia virus. Hamsters vaccinated with the recombinant gp13 vaccinia virus survived a lethal challenge with the hamster-adapted Kentucky strain of EHV-1. These results indicate that expression in vaccinia virus vectors of EHV-1 gp13, the glycoprotein homolog of herpes simplex virus gC-1 and gC-2, pseudorabies virus gIII, and the varicella-zoster virus gpV may provide useful vaccine candidates for equine herpesvirus infections. PMID- 2550664 TI - Large T-antigen mutants define multiple steps in the initiation of simian virus 40 DNA replication. AB - The biochemical activities of a series of transformation-competent, replication defective large T-antigen point mutants were examined. The assays employed reflect partial reactions required for the in vitro replication of simian virus 40 (SV40) DNA. Mutants which failed to bind specifically to SV40 origin sequences bound efficiently to single-stranded DNA and exhibited nearly wild-type levels of helicase activity. A mutation at proline 522, however, markedly reduced ATPase, helicase, and origin-specific unwinding activities. This mutant bound specifically to the SV40 origin of replication, but under certain conditions it was defective in binding to both single-stranded DNA and the partial duplex helicase substrate. This suggests that additional determinants outside the amino terminal-specific DNA-binding domain may be involved in nonspecific binding of T antigen to single-stranded DNA and demonstrates that origin-specific DNA binding can be separated from binding to single-stranded DNA. A mutant containing a lesion at residue 224 retained nearly wild-type levels of helicase activity and recognized SV40 origin sequences, yet it failed to function in an origin-specific unwinding assay. This provides evidence that origin recognition and helicase activities are not sufficient for unwinding to occur. The distribution of mutant phenotypes reflects the complex nature of the initiation reaction and the multiplicity of functions provided by large T antigen. PMID- 2550666 TI - Treatment of encephalomyocarditis virus-induced central nervous system demyelination with monoclonal anti-T-cell antibodies. AB - Infection of BALB/c mice with the M variant of encephalomyocarditis virus resulted in the development of a paralytic syndrome in 7 to 10 days. The paralysis was maximal during the period of viral clearance; most of the animals recovered from the initial deficit and showed no delayed recurrences. Pathologically, the white matter of brain and spinal cord showed well-demarcated areas of perivascular cuffing, demyelination, and, during recovery, remyelination by oligodendrocytes--all suggestive of postinfectious encephalomyelitis. Depletion of either the CD4 or CD8 subset of T cells in vivo with the appropriate monoclonal antibody, GK1.5 or 2.43, respectively, administered one day (24 h) prior to infection was sufficient to limit the development of the paralytic syndrome by 79% (GK1.5) and 82% (2.43). PMID- 2550667 TI - Varicella-zoster virus glycoprotein oligosaccharides are phosphorylated during posttranslational maturation. AB - Varicella-zoster virus (VZV)-infected human embryonic lung fibroblasts (HELF) do not release infectious virions into their growth medium. Extracellular virions are pleomorphic, suggesting that they are partially degraded before their release from cells. To examine the intracellular pathway of viral maturation, [2 3H]mannose-labeled virus-encoded glycoproteins were isolated from VZV-infected HELF. Oligosaccharides attached to the glycoproteins were processed to complex type units, some of which were phosphorylated. The major intracellular site of accumulation of VZV gpI was found to be perinuclear and to correspond to that of the cation-independent mannose 6-phosphate (Man 6-P) receptor. Subsets of VZV containing cytoplasmic vacuoles were coated, Golgi-associated, or accessible to endocytic tracers. Phosphorylated monosaccharides protected HELF from the cytopathic effect of VZV in proportion to their ability to block Man 6-P receptor mediated endocytosis. These data suggest that the unusual phosphorylated oligosaccharides mediate an interaction between VZV and Man 6-P receptors of the host cell; this interaction may be responsible for withdrawal of newly synthesized virions from the secretory pathway and for their diversion to prelysosomal structures. PMID- 2550668 TI - Cytocidal effect caused by the envelope glycoprotein of a newly isolated avian hemangioma-inducing retrovirus. AB - We isolated a field strain of avian hemangioma retrovirus (AHV) which induces a cytopathic effect (CPE) on cultured avian and mammalian cells shortly after infection. The kinetics of cell killing were dependent on the multiplicity of infection. The CPE on avian and mammalian cells was independent of virus replication, because UV-irradiated virus led to cell death as well. Biochemical and genetic experiments indicated that AHV env gene products were responsible for the CPE. Partially purified AHV envelope glycoproteins (gp85), but not those of the Rous sarcoma virus Prague C strain, induced a CPE. Rous-associated virus type 1, in which the env region was replaced by the AHV gp85 region, induced a CPE on avian and mammalian cultured cells. Therefore, we suggest that CPE is induced by AHV via interaction between viral gp85 and the cell membrane. This mode of CPE is unique among avian sarcoma-leukemia viruses. PMID- 2550669 TI - Creation and expression of myristylated forms of Rous sarcoma virus gag protein in mammalian cells. AB - Rous sarcoma virus (RSV), a member of the avian sarcoma and leukosis family of retroviruses, has long been known to be capable of infecting and transforming mammalian cells; however, such transformed cells do not release virus particles. The RSV gag product (Pr76gag) produced in these cells is not released into the culture medium or proteolytically processed to release mature products. Thus, the behavior of Pr76gag in mammalian cells is much like that of mammalian retroviral Gag proteins which have been altered so as to block the addition of myristic acid at residue 2 (Gly). Because the RSV gag product does not possess a myristic acid addition site, we hypothesized that the creation of one by oligonucleotide directed mutagenesis might permit particles to be released from mammalian cells. Two myristylated forms of Pr76 were created. In Pr76myr1, the first 10 amino acids have been exchanged for those of p60v-src, which are known to be sufficient for myristylation. In Pr76myr2, the Glu at the second residue has been substituted with Gly. The alleles encoding the modified and wild-type forms of Pr76 have been expressed at high levels in mammalian (CV-1) cells by using an SV40-based vector. Surprisingly, we have found that expression of high levels of the unmodified (wild-type) product, Pr76myr0, results in low levels of particle formation and precursor processing. This indicates that myristic acid is not the sole determinant for targeting. However, the addition of myristic acid to Pr76myr1 or Pr76myr2 resulted in a fivefold enhancement in Gag function. In all aspects examined, the behavior of myristylated Pr76 was identical to that of the authentic product produced in avian cells. We also show that processing is mediated by the gag-encoded protease and that removal of the amino terminus to create Pr76gagX results in an inability to form particles or be processed. This suggests that proper targeting is prerequisite for activation of the RSV protease in mammalian cells. PMID- 2550670 TI - Age-dependent poliomyelitis of mice: expression of endogenous retrovirus correlates with cytocidal replication of lactate dehydrogenase-elevating virus in motor neurons. AB - The widespread presence of endogenous retroviruses in the genomes of animals and humans has suggested that these viruses may be involved in both normal and abnormal developmental processes. Previous studies have indicated the involvement of endogenous ecotropic murine leukemia virus (MuLV) in the development of age dependent poliomyelitis caused by infection of old C58 or AKR mice by lactate dehydrogenase-elevating virus (LDV). The only genetic components which segregate with susceptibility to LDV-induced paralytic disease are multiple proviral copies of ecotropic MuLV and the permissive allele, at the Fv-1 locus, for N-tropic, ecotropic virus replication (Fv-1n/n). Using in situ hybridization and Northern (RNA) blot hybridization, we have correlated the expression of the endogenous MuLV, both temporally and spatially, with LDV infection of anterior horn motor neurons and the development of paralysis. Our data indicate that treatment of 6- to 7-month-old C58/M mice with cyclophosphamide, which renders these mice susceptible to LDV-induced paralytic disease, results in transient increases in ecotropic MuLV RNA levels in motor neurons throughout the spinal cord. Peripheral inoculation of C58/M mice with LDV, at the time of elevated MuLV RNA levels, results in a rapid spread of LDV to some spinal cord motor neurons. LDV infections then spread slowly but progressively throughout the spinal cord, involving an increasing number of motor neurons. LDV replication is cytocidal and results in neuron destruction and paralysis of the infected animals 2 to 3 weeks postinfection. The slow replication of LDV in the spinal cord contrasts sharply with the rapid replication of LDV in macrophages, the normal host cells for LDV, during the acute phase of infection. The data indicate that the interaction between the endogenous MuLV with the generally nonpathogenic murine togavirus LDV occurs at the level of the motor neuron. We discuss potential mechanisms for the novel dual-virus etiology of age-dependent poliomyelitis of mice. PMID- 2550671 TI - Characterization of in vitro inhibition of human immunodeficiency virus by purified recombinant CD4. AB - The first step in infection of human T cells with human immunodeficiency virus (HIV) is binding of viral envelope glycoprotein gp120 to its cellular receptor, CD4. The specificity of this interaction has led to the development of soluble recombinant CD4 (rCD4) as a potential antiviral and therapeutic agent. We have previously shown that crude preparations of rCD4 can indeed block infection of T cells by HIV type 1 (HIV-1). Here we present a more detailed analysis of this antiviral activity, using HIV-1 infection of the T lymphoblastoid cell line H9 as a model. Purified preparations of rCD4 blocked infection in this system at nanomolar concentrations; combined with the known affinity of the CD4-gp120 interaction, this finding suggests that the inhibition is simply due to competition for gp120 binding. As predicted, rCD4 had comparable activity against all strains of HIV-1 tested and significant activity against HIV-2. Higher concentrations of rCD4 blocked infection even after the virus had been adsorbed to the cells. These findings imply that the processes of viral adsorption and penetration require different numbers of gp120-CD4 interactions. Recombinant CD4 was able to prevent the spread of HIV infection in mixtures of uninfected and previously infected cells. Our studies support the notion that rCD4 is a potent antiviral agent, effective against a broad range of HIV-1 isolates, and demonstrate the value of purified rCD4 as an experimental tool for studying the mechanism of virus entry into cells. PMID- 2550672 TI - Simian virus 40 late transcripts lacking excisable intervening sequences are defective in both stability in the nucleus and transport to the cytoplasm. AB - Little or no simian virus 40 (SV40) late mRNA accumulates in the cytoplasm when the primary transcript lacks an excisable intervening sequence. To begin to understand why, we analyzed the synthesis, processing, transport, and stability of SV40 late transcripts accumulated in the nucleus and cytoplasm of monkey cells cotransfected with the DNAs of wild-type and mutants of SV40 lacking precisely various introns. The data from these experiments indicated that (i) the presence of excisable intervening sequences in SV40 late transcripts is necessary for efficient accumulation in the cytoplasm of any of the SV40 late RNA species and (ii) SV40 late transcripts lacking excisable intervening sequences are defective in both stability in the nucleus and transport to the cytoplasm but not in stability in the cytoplasm. We hypothesize that SV40 late transcripts need to be processed via a pathway that couples stabilization of the primary transcript within the nucleus, excision of intervening sequences, proper 5'- and 3'-end formation, and transport to the cytoplasm. PMID- 2550673 TI - Trans activation by the bovine papillomavirus E2 protein in Saccharomyces cerevisiae. AB - The papillomavirus E2 protein functions as an enhancer-binding factor to promote transcription in mammalian cells. We found that one copy of the E2 binding site acted as an E2 protein-dependent upstream activating sequence in Saccharomyces cerevisiae. Additional copies of the binding motif further augmented transcription. These results imply that the E2 protein functionally interacts with highly conserved transcriptional elements. PMID- 2550674 TI - Functional significance of phosphorylation to the human immunodeficiency virus Rev protein. AB - The human immunodeficiency virus Rev protein is posttranslationally modified by a serine kinase activity present in the nucleus of the cell. Site-directed mutagenesis was used to identify the site of phosphorylation. Changing of serine residues 92 and 99 dramatically reduced Rev phosphorylation, suggesting that at least one, if not both, of these residues is the one recognized by the Rev specific serine kinase. Similarly, a truncated Rev protein lacking the 25 carboxy terminal amino acids was not phosphorylated. By using two independent assays, both the serine mutant proteins and the truncated form of Rev were found to be fully functional. Thus, phosphorylation and the 25 carboxy-terminal amino acids appear to be dispensable for protein function. PMID- 2550675 TI - Guanidine-resistant mutants of poliovirus have distinct mutations in peptide 2C. AB - In previous work in our laboratory, 12 guanidine-resistant (gr) mutants of poliovirus were selected from 12 separate stocks of plaque-purified guanidine sensitive (gs) viruses (K. Anderson-Sillman, S. Bartal, and D. R. Tershak, J. Virol. 50:922-928, 1984). Peptide mapping of protein 2C and evaluation of virus growth at different temperatures enabled us to subdivide these mutants into several distinct groups (D. R. Tershak, Can. J. Microbiol. 31:1166-1168, 1985; Anderson-Sillman et al., J. Virol.). Studies by Pincus et al. indicate that two nucleotide changes in codon 179 of protein 2C leads to an Asn-to-Gly or Asn-to Ala change and that these missense modifications account for guanidine resistance (S. E. Pincus, H. Rohl, and E. Wimmer, Virology 157:83-88, 1987; S. E. Pincus and E. Wimmer, J. Virol. 60:793-796, 1986). In the present report, we confirm their findings but also show that a single amino acid change of Phe to Tyr in residue 164 of protein 2C or a Met-to-Leu replacement in amino acid 187 coupled with mutations in codons 233 or 295 and 309 could confer guanidine resistance. PMID- 2550676 TI - Direct isolation and characterization of JC virus from urine samples of renal and bone marrow transplant patients. AB - JC virus DNA was extracted from urine-derived cells of bone marrow and renal transplant patients and cloned directly into the plasmid vector pBR322. These clones represent the first JC virus isolates obtained directly from individuals that did not have progressive multifocal leukoencephalopathy (PML). Three of the clones appeared to be identical to the prototype JC virus Mad 1, and the fourth clone was identical to the type II JC virus variant Mad8-Br. Importantly, the same JC virus strains have been identified both in the urine of non-PML patients and in the brain tissue of PML patients. These results indicate that different organs may be infected with the same JC virus subtype and implies that an adaptation process involving the alteration of viral regulatory signals is not required in the pathogenesis of PML. Furthermore, both a type I and a type II variant were obtained from the same patient, suggesting that an individual may be infected with more than one strain of JC virus at a given time. PMID- 2550677 TI - Expression from the adeno-associated virus p5 and p19 promoters is negatively regulated in trans by the rep protein. AB - The leftward two promoters of the adeno-associated virus (AAV) 2 genome were fused to reporter genes, and the constructs were used to transfect HeLa cells. The promoters functioned constitutively but were repressed in trans by the AAV rep gene product(s). The repression was relieved by adenovirus infection. Evidence which indicated an enhancer function for the inverted terminal repeat of the AAV-2 genome was also obtained. PMID- 2550678 TI - Herpes simplex virus type 1 latency-associated transcription plays no role in establishment or maintenance of a latent infection in murine sensory neurons. AB - Using herpes simplex viruses deleted and restored for the latency-associated transcripts (LATs), we have quantitatively assessed the role of the transcripts in establishment and maintenance of latent infection. Determination of the number of neurons latently infected and the copy number of viral genomes per latently infected ganglion indicated that there is no difference between viruses expressing and not expressing the transcripts. In addition, the amount of viral DNA present in ganglia latently infected with the LAT-negative virus KOS 8117 did not differ from the value for LAT+ counterparts over an 11-month period of analysis. From these results we conclude that LATs play no role in establishment or maintenance of a latent infection with herpes simplex virus type 1. If these transcripts play a role in latency, they must function during the reactivation step. PMID- 2550680 TI - The Albert Lasker Medical Awards. Role of the cyclic AMP-dependent protein kinase in signal transduction. AB - The first example of an enzyme that undergoes phosphorylation and dephosphorylation was phosphorylase, which catalyzes the initial step in the breakdown of tissue glycogen, a process that occurs when cells are stimulated by certain hormones. The phosphorylation reaction that activates phosphorylase is catalyzed by an enzyme designated "phosphorylase kinase," whereas the dephosphorylation reaction is catalyzed by an enzyme called "phosphorylase phosphatase." Phosphorylase kinase, like phosphorylase itself, is regulated by phosphorylation-dephosphorylation. In this instance the enzyme is phosphorylated and activated by a protein kinase that is the intracellular target of adenosine 3',5'-cyclic phosphate, the second messenger of hormone action that was discovered by the late Earl Sutherland. Knowledge of these steps led to the establishment of the glycogenolytic cascade in which adenosine 3',5'-cyclic phosphate, formed as a result of hormone action, first activates the adenosine 3',5'-cyclic phosphate-dependent protein kinase, which in turn activates phosphorylase kinase, which then converts inactive phosphorylase to its active form. This is followed by the breakdown of glycogen. PMID- 2550679 TI - Effects of mutations in hyperconserved regions of the extracellular glycoprotein of human immunodeficiency virus type 1 on receptor binding. AB - Sequence comparison of the human immunodeficiency virus type 1 and type 2 env genes revealed the presence of six linear regions in the extracellular glycoprotein that are highly conserved. To investigate the functional significance of these regions, we made short deletions in each and assayed the ability of the mutated proteins to bind CD4 antigen. Small deletions in four of the highly conserved regions drastically reduced receptor binding. Some deletions interfered with the maturation of the envelope glycoprotein, but maturation did not necessarily correlate with the ability to bind CD4 antigen. PMID- 2550681 TI - [Estrogen and androgen receptors in hepatocellular carcinoma and in noncancerous liver tissue]. AB - We have assayed both estrogen and androgen receptors in hepatocellular carcinomas, in the surrounding noncancerous liver tissue, and in normal liver tissue. Estrogen receptors (ERs) were detected in 5 out of 22 HCC tissues, 14 out of 22 surrounding noncancerous tissues, and in 5 out of 6 normal liver tissues. Androgen receptors (Ars) were detected in 4 out of 21 HCC tissues, in 7 out of 21 surrounding noncancerous tissues, and in 2 out of 5 normal liver tissues. These results suggest that the pathogenesis of some HCC may be dependent on estrogen or androgen. Hence, it may be beneficial to provide hormonal therapy for HCC, especially in cases that show high concentrations of ER and/or AR. PMID- 2550682 TI - [A clinicopathological study of early gastric cancers with a diameter larger than five centimeters]. AB - Out of a total of 1,112 resected early gastric carcinomas, 181 that were larger than 5 cm in diameter have been pathologically investigated. Of these, intramucosal and submucosal carcinomas amounted to 68 (37.6%) and 113 (64.4%), respectively. The incidence of their location, shown as a percentage, was 37.8% in the antrum, 57.4% in the corpus, and 4.8% in the fundus (11% in the anterior wall, 13.2% in the posterior wall, 68.1% in the lesser curvature, and 7.7% in the greater curvature). Grossly, the incidence of a type IIc carcinoma was 46.5% and that of a IIc + III type was 20.5%, respectively. Microscopically, in the intramucosal cases, signet ring cell carcinomas were the most frequent histological type, whereas in the invasive submucosal cases, the carcinomas were the intestinal metaplastic type. Lymphatic invasions, venous invasions, and lymph nodal metastases amounted to 32.6%, 6.1%, and 11.6%, respectively. In the early gastric carcinomas, the larger the tumor size, the more likelihood of a signet ring cell carcinoma than a intestinal metaplastic type, and it appeared that a signet ring cell carcinoma had infiltrated the propria mucosae for a longer time when compared to either an intestinal metaplastic type carcinoma or a poorly differentiated tubular adenocarcinoma. PMID- 2550683 TI - [The correlation between preoperative pathologic diagnosis of a biopsy specimen and postoperative pathologic diagnosis of a tissue specimen involving colorectal cancer patients]. AB - An endoscopic biopsy was performed on specimens taken from 374 patients with a large bowel cancer, who had received a colo-rectal resection between 1980 and 1984. An average of three to four biopsy specimens was taken from each patient. The pathological diagnosis of these specimens revealed a carcinoma with an identifiable pathological classification in 280 patients (74.9%), a carcinoma without an identifiable classification in 54 (14.4%), a suspect cancer in 7 (1.9%), and no sign of a cancer in 33 (8.8%). The diagnosis of the 33 cases in which no cancer was detected were adenoma in 11 patients, inflammation in 8, necrosis in 1, and no clear pathological determination in 11. The correspondence rates between a biopsy specimen and a tissue specimen diagnoses were 57.9% in a well differentiated adenocarcinoma; 77.4% in a moderately differentiated adenocarcinoma; 85.5% in a poorly differentiated adenocarcinoma; and 100% in a mucoid carcinoma. PMID- 2550684 TI - [The accuracy of CT and tumor markers in the detection of a recurrent ovarian carcinoma]. AB - Twenty-three patients previously diagnosed as having ovarian cancer were examined with both serum tumor markers (CA 125, CA 19-9, TPA, IAP, AFP) and a pelvic CT scan. The tumor markers predict the clinical outcome more accurately than the CT scan. Further, the tumor markers showed a clear correlation with the clinical course. But in one case, however, the tumor markers were seen to reduce below the normal level from chemotherapy, while the CT scan showed a tumor mass. Thus, both, a CT scan and tumor marker assays are felt to be indispensable for detecting the recurrence of an ovarian cancer. PMID- 2550685 TI - [Occurrences of human papilloma virus DNA in cervical carcinomas]. AB - Out of 16 cases involving a cervical carcinoma that were investigated by Southern blot hybridization, found were human papilloma virus (HPV) types 16 and 18 DNA sequences in 8 (50%) and in one (6.3%), respectively. Six out of the 8 HPV 16 positive specimens were from squamous cell carcinomas, one was from an adenocarcinoma, and the remaining specimen was from an argyrophil small cell carcinoma. In 7 out of 9 HPV-positive specimens, the viral sequences were integrated in the tumor cell genome, whereas in the remaining two they were not integrated and remained circular and/or oligomeric in form. PMID- 2550686 TI - [Radiotherapy in bone metastases--with special reference to its effect on relieving pain]. AB - Between December 1986 and January 1978, 68 patients with bone metastases were analyzed to evaluate the effect of radiation for the relief of pain. The 68 patients, who had a total of 97 lesions, complained of pain caused by their bone metastasis. The good, fair, and poor responses were found to be 18%, 60%, and 22%, respectively. With reference to the primary neoplasms, the effective response rate was 73% in lung cancer, 100% in breast cancer, 75% in gastric cancer, 100% in hepatic cancer, 100% in bladder cancer, 25% in epipharyngeal cancer, and 70% in the other neoplasms. Depending on the cell types of the lung cancer, the effective response rate was 80% for small cell carcinomas, 72% for adenocarcinomas and 40% for squamous cell carcinomas. Our results suggest that radiotherapy for bone metastases is to be recommended, since the effective response rate was 78% for the relief of pain. PMID- 2550687 TI - [A case of Zollinger-Ellison syndrome successfully treated with an H+-K+ ATPase inhibitor]. AB - A case of Zollinger-Ellison syndromes in a fifty year-old male that was successfully treated with a H+-K+ ATPase inhibitor (Omeprazol) is reported. The patient underwent a partial gastrectomy in 1984, but had been suffering from multiple refractory stomal and jejunal ulcers after the operation. In 1987, hypergastrinemia (760 pg/ml) was detected, and the presence of gastrinomas in the pancreatic head accompanied by a multiple liver metastasis was subsequently confirmed by CT-angiography and by the gastrin level detected in percutaneous, transhepatic, portal venous samples. A secretin provocation test proved to be negative, and the ulcers resisted the H2-receptor antagonists, but the patient was successfully cured shortly after the administration of an H+-K+ ATPase inhibitor. PMID- 2550688 TI - [Three cases of vulvar bowenoid papulosis: the localization of HPV DNA by in situ hybridization]. AB - Cytological, histological, and molecular biological studies were conducted in 3 cases of vulvar Bowenoid papulosis, using biotinylated HPV DNA probes by in situ hybridization. 1) Cytological findings showed dyskaryotic cells that revealed hyperchromatism with a coarse granular pattern, and a high N/C ratio was observed among the dyskeratotic cells. 2) In 2 cases of Bowenoid papulosis lesions, HPV 16 DNA was detected in the nucleus of the dysplastic cells. 3) In one case of Bowenoid papulosis, a complicated carcinoma in situ of the uterine cervix was observed, and the HPV 16 DNA was found to be positive in both the vulva and cervix. PMID- 2550689 TI - Varicella-zoster virus prevalence in Japan: no significant change in a decade. AB - A seroepidemiologic time-comparison study was conducted to evaluate changes in IgG antibody to varicella-zoster virus (VZV) and to determine VZV prevalence in Japan with randomly collected serum samples from two healthy Japanese populations: 1973 (n = 670) vs. 1984 (n = 677). Enzyme-linked immunosorbent assay (ELISA) was found to be superior to the immune adherence hemagglutination test (IAHA) especially for detecting seropositivity in adults. Serologic results showed essentially no significant difference between the 1973 and the 1984 age specific prevalences; with the exception of a slightly lower prevalence in the 5 year-old age group in 1973 than that in 1984. In general, the age-specific prevalence was lowest in the 1-year-old group (mean 11%) and increased in a linear fashion until age 9 (mean 82.9%); prevalence continued to increase slowly after age 9 and plateaued at 100% for ages greater than or equal to 25-29. PMID- 2550690 TI - Effect of l-ephedrine on cholinergic neurotransmission in the isolated guinea pig ileum. AB - In the isolated guinea pig ileum, the effects of I-ephedrine on the twitch response to field stimulation were investigated in the presence of propranolol. Ephedrine and clonidine inhibited the twitch response but not the contraction to exogenous acetylcholine. The inhibitory effect of clonidine was significantly diminished by yohimbine pretreatment. However, the inhibitory effect of ephedrine was not influenced by yohimbine, sulpiride or 6-hydroxydopamine (6-OHDA) pretreatment. Most of this action of ephedrine appeared to be cholinergic prejunctional in nature, but unrelated to activation of prejunctional alpha 2 adrenoceptors and dopamine sensitive receptors on the cholinergic nerves in this preparation. PMID- 2550692 TI - Abnormal plasma prothrombin (PIVKA-II) levels in hepatocellular carcinoma. AB - The concentration of abnormal prothrombin, or the protein induced by vitamin K absence or antagonist II (PIVKA-II) in 102 patients with hepatic disorders was measured by an enzyme immunoassay method. The concentration of PIVKA-II in the plasma was elevated in 11 out of 18 patients with hepatocellular carcinoma and also in a patient with hepatoblastoma. There was no correlation between serum alpha-fetoprotein and plasma PIVKA-II levels. The PIVKA-II level was normal in 11 patients who had metastatic carcinoma or cholangiocellular carcinoma. Moreover, benign diseases of the liver did not cause an elevation in PIVKA-II. PIVKA-II might be an useful marker of hepatocellular carcinoma because, like alpha fetoprotein, its level changes in close relation to the effects of treatment. PMID- 2550691 TI - Malignant fibrous histiocytoma in the thoracic region--a clinico-pathologic investigation. AB - Seven cases of malignant fibrous histiocytoma (MFH); 3 originating in the thoracic region, one which was considered to arise from the lung parenchyma, and 4 which were metastatic to the lungs, are presented herein. Six of these patients underwent surgical excision and analysis under light and electron microscopy revealed the lesions of MFH to be composed of two cell types; a fibroblast-like cell and a histiocyte-like cell. The latter showed histologically characteristic growth in a so-called storiform pattern. In all patients adjuvant chemotherapy was performed, although in only one patient did it prove temporarily effective. Despite the fact that the prognosis of MFH in the thoracic region is poor, the suggested therapy for longer survival is resection with postoperative combination chemotherapy including the use of sensitive anticancerous agents. PMID- 2550693 TI - Activity of calcium-sensitive phospholipid-dependent protein kinase C following nephron loss. AB - The localization and activity of the calcium-sensitive phospholipid-dependent protein kinase C (PKC) were examined following the loss of 50% of functioning nephron mass. Four hours following unilateral nephrectomy in rats, soluble (100,000 g supernatant) proteins in the contralateral kidney were increased by 11% compared to sham operated controls; the increase was 33% 144 hours following surgery. The specific activity of PKC did not change in the cytosol at any of the time periods examined and averaged 63.9 +/- 8.2 pmol.mg-1.min-1 in unilaterally nephrectomized animals four hours following surgery. Four hours following sham surgery total soluble PKC activity averaged 1667.0 +/- 278.4 pmol.kidney-1.min-1, whereas activity averaged 3067.7 +/- 415.4 pmol.kidney-1.min-1 in animals post nephrectomy (N = 5, P less than 0.04). Similar data was seen 144 hours following surgery. To examine the PKC activity in plasma membranes of proximal tubular cells, brush border membranes were prepared from rat kidney cortex. Twenty-four hours following unilateral nephrectomy, activity averaged 193.8 +/- 14.9 pmol.mg 1.min-1, while activity in membranes isolated from sham operated animals averaged 76.6 +/- 8.0 pmol.mg-1.min-1 (N = 5, P less than 0.001). Similar data was evident 48 hours following surgery. A small increment in activity was seen in the basolateral membrane preparation 24 hours following unilateral nephrectomy but not at 48 hours. These data indicate that cellular PKC activity increases rapidly following reductions in renal mass, and there are selective increments in the brush border membrane of the proximal tubular cell. The localization of PKC to this membrane may have important consequences for adaptations following nephron loss. PMID- 2550694 TI - Renal mesangium is a target for calcitonin gene-related peptide. AB - Rat calcitonin gene-related peptide (CGRP alpha; EC50, 1 nM) was shown to stimulate cAMP formation in cultured rat renal mesangial cells. CGRP concentration dependently (EC50, 1 nM) also inhibited contraction of mesangial cells by angiotensin II (10 nM). Angiotensin II (10 nM) caused a transient increase of the intracellular calcium concentration from 140 nM to 480 nM in the mesangial cells, but these calcium transients were not altered by CGRP. CGRP (10 nM) decreased vascular resistance in the isolated rat kidney perfused at constant pressure (100 mm Hg; P less than 0.01). The decreased vascular resistance was accompanied by a rise of the glomerular filtration fraction. CGRP, moreover, attenuated the effects of angiotensin II on renal vascular resistance and glomerular filtration (P less than 0.01). In conclusion, CGRP causes relaxation of renal mesangial cells and decreases renal vascular resistance. As a result CGRP raises glomerular filtration and the filtration fraction. The effect may be linked to cyclic AMP formation. Thus, regulation of renal vascular and glomerular function may represent a novel action of CGRP apart from its cardiovascular effects. PMID- 2550695 TI - Macrophages selectively stimulate ecto-5'-nucleotidase activity of cultured mesangial cells. AB - Because rat peritoneal macrophages exhibit a particular binding capacity to rat glomerular mesangial cells in vitro, we have studied the effect of macrophages on two plasma membrane enzymes, ecto-5'-nucleotidase and ecto-Mg2+ ATPase, or rat cultured mesangial cells. A marked increase of ecto-5'-nucleotidase activity was observed in cocultures of rat mesangial cells and peritoneal macrophages. In addition, macrophage-conditioned medium (MCM) produced a dose-dependent increase of mesangial cell ecto-5'-nucleotidase activity. In contrast, ecto-Mg2+ ATPase activity was unaffected. The effect of MCM on ecto-5'-nucleotidase activity was apparent after 24 hours and increased with time. Reversion was obtained by MCM withdrawal. Stimulation of ecto-5'-nucleotidase activity by MCM was inhibited by cycloheximide, which suggests that protein synthesis was required. Induction of enzyme activity by MCM depended in part on the presence of extracellular adenosine. The macrophage-released factor responsible for this effect was non dialysable, heat-stable at 56 degrees C for 30 minutes but heat-inactivated at 100 degrees C for five minutes, inactivated in the presence of trypsin or protease V8, and adsorbed on charcoal. Its apparent molecular weight estimated by gel chromatography was close to 20 kD. MCM from resident macrophages was as potent as MCM from thioglycollate-elicited or zymosan-stimulated macrophages. This stimulatory effect was specific of macrophages since it was absent in the culture medium of rat fibroblasts or mouse thymocytes but present in that of mouse macrophages. These results suggest that peritoneal macrophages synthesize a factor which selectively stimulates ecto-5'-nucleotidase activity of glomerular mesangial cells. PMID- 2550697 TI - [Definition of malignant histiocytosis and large cell anaplastic (Ki-1) lymphoma in children]. AB - Cells belonging to the mononuclear phagocytic system may give rise to a wide variety of proliferative disorders. These neoplastic or reactive diseases of monocytes, histiocytes and macrophages are still poorly understood, both from a biological, clinical and pathological point of view. In 1984 a new type of non Hodgkin's lymphomas has been described, which recently was termed large cell anaplastic lymphoma (LCAL). Since both "true" malignant histiocytosis (MH) and LCAL display some clinico-pathological similarities, we reevaluated eleven children with the primary diagnosis of MH, who entered the multicentric therapy study DAL-HX 83. On the basis of the typical morphology LCAL and not MH has been diagnosed in all cases. The Ki-1 (CD 30) positive tumor cells of LCAL can display different phenotypes, either T-lymphoid, B-lymphoid, histiocytic or neither of them (O-phenotype). Therefore we investigated the lineage specificity of tumor cells in 15 paediatric patients with LCAL. Tumor cells of histiocytic origin could be demonstrated in only one of them. In 7 patients the neoplastic cells were of T cell and in the remaining 7 cases of non T/non B cell (O) phenotypes, confirming the extreme rarity of MH in children. It is our opinion that MH and LCAL seem to be different diseases not only nosologically but also clinico pathologically. In the multicentric therapy studies NHL-BFM 81 and NHL-BFM 83 the frequency of LCAL in childhood is compared to all other subentities at 3%. PMID- 2550696 TI - Efficacy and variability of the antiproteinuric effect of ACE inhibition by lisinopril. AB - We studied the efficacy of the ACE inhibitor lisinopril in treating overt proteinuria in comparison with the NSAID indomethacin, and evaluated some of the conditions that could influence this antiproteinuric effect. In 12 patients with a proteinuria varying from 3.2 to 10.5 g/24 hr, a diastolic BP ranging from 64 to 105 mm Hg, and a GFR varying from 34 to 127 ml/min, the effect of different lisinopril doses and of changing dietary sodium intake was evaluated. Proteinuria fell by 27 +/- 20% from 6.1 +/- 2.1 to 4.5 +/- 1.9 g/24 hr on a low dose (median 5 mg/day) lisinopril and by 50 +/- 17% to 3.1 +/- 1.4 g/24 hr on a higher dose (median 10 mg/day), irrespective of initial proteinuria, BP, or GFR. This antiproteinuric effect was abolished by increasing salt intake from 50 to 200 mmol/day, and was recovered again by re-instituting the sodium restricted diet. The antiproteinuric effect of 10 mg/day lisinopril was comparable to the reduction in proteinuria (by 57 +/- 21% to 2.8 +/- 2.0 g/24 hr) on 150 mg/day indomethacin, while adverse effects were less and renal hemodynamic effects were more favorable during lisinopril. In some patients it took several weeks before the effect of the ACE inhibitor on proteinuria was stabilized. Thus, the antiproteinuric effect of the ACE inhibitor lisinopril appears to be dose and time related, and is strongly dependent on dietary sodium restriction, whereas it does not depend on initial proteinuria, BP, or GFR. The effect is comparable to that of indomethacin, while adverse effects are less. PMID- 2550698 TI - [Large cell anaplastic lymphoma in children--clinical experiences with a newly defined histologic entity]. AB - The clinical characteristics of 31 patients (pts.) (17 boys, 14 girls, median age 12 11/12 years) with large cell anaplastic lymphoma (LCAL) have been evaluated. 17 of these pts. had originally been diagnosed as suffering from "malignant histiocytosis" ("MH") and were therefore included in the DAL-Histiocytosis X 83 study. Another 14 pts. with Ki-1 lymphomas were enrolled in the BFM-NHL therapy studies. According to Murphyclassification 24 pts. (77%) had stage III or IV disease and in general presented in a severe condition. The lymphatic system was involved in 28 pts., 8 pts. (26%) had skin infiltration. With regard to lymphoma involvement of lung, bones and bone-marrow were unexpectedly frequent. CNS involvement was seen in just one pt. Despite rather heterogeneous therapy approaches (ALL-schedules, DAL-HX 83 protocol for treatment of "MH", combination of B-NHL-BFM and AML-BFM schedules, CHOP, BFM protocols for B-NHL) 30 out of 31 pts. achieved clinical remission (CR). The only nonresponder died during bone marrow transplantation of septicemia. 4 pts. relapsed during therapy. 3 of them died, 1 during a BMT. 1 pt. achieved 2nd CR with a BFM-B-NHL protocol. 3 pts. experienced a late relapse, 1 died, 1 2nd CR was achieved, the third pt. is still alive after 2 further relapses disease-free for 3 years. 23 pts. (74%, 13 out of 14 of BFM-NHL therapy study, 10 out of 17 of DAL-HX 83 study, 1 pt. after BMT) are in 1st CR with a median observation time of 2 9/12 years (range 5/12 to 17 9/12 years).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550699 TI - [Cytogenetic findings in Wilms' tumor]. AB - We report the cytogenetic analysis of 20 Wilms'tumors. In 10 cases chromosomal abnormalities were found: Besides the expected aberrations involving 11p13 we found chromosome 11 breakpoints at 11p15, 11q13 or q23. Chromosome 1 was frequently involved in rearrangements, with partial trisomy 1q as the typical abnormality. Additional chromosomes 8, 12, 13, 17 and 20 were the most frequent numerical changes. This study is part of efforts to find cytogenetic and molecular markers and constellations contributing important information beyond histology and DNA-measurements. PMID- 2550700 TI - [Prevention of cytomegalic inclusion disease in leukemia patients with a cytomegalovirus hyperimmune globulin preparation]. AB - In a randomised study the efficacy of a cytomegalic hyperimmune globulin preparation (CMV-HIGP) which had been treated with beta-propiolactone was analysed. The study included 85 patients with acute lymphoblastic leukemia (ALL) and Non-B-Non-Hodgkin-lymphoma (NHL) who were treated initially or underwent a relapse therapy. During the intense chemotherapeutical period within leukemia treatment the patients were passively immunised by the intravenous route with CMV HIGP (1 ml per kilogram of body weight) every two to three weeks at the latest. In the initial stages the basic immunisation protection was achieved by the application of double dose CMV-HIGP. The Frankfurt patients were recruited from the BFM-ALL- and the NHL-study since october 1982. When they were admitted their CMV serostatus was determined by means of the ELA-ELISA or IFA-method. Seronegative patients were given the passive immunisation immediately or 48 hours after the first blood transfusions at the latest. The patients who had become CMV IgG-positive by passive immunisation were randomised when reaching long-term therapy according to the protocol. Because of a 30% cytomegaly disease incidence rate in our patient population a randomisation was unwarrantable at the beginning of leukemia treatment. During randomisation one group of patients were immunised by the intravenous route with CMV-HIGP (2 ml per kg body weight one time in four weeks), the second group was a control group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550701 TI - [Myeloperoxidase of neutrophils and its possible role in lipid peroxidation processes in arteriosclerosis]. AB - Activity of blood plasma myeloperoxidase (MPO) of neutrophil leucocytes and acetyl hydroperoxides was studied in families of atherosclerosis patients. The neutrophil MPO activity was decreased and the blood plasma content of acetyl hydroperoxides was increased in subjects with hereditary predisposition to atherosclerosis. The role of MPO and its possible pathogenetic action as a generator of the active forms of oxygen in atherosclerosis and its complications has been discussed. PMID- 2550702 TI - Urinary excretion half-life of delta 1-tetrahydrocannabinol-7-oic acid in heavy marijuana users after smoking. AB - The urinary excretion of the total amount of delta 1-tetrahydrocannabinol (delta 1-THC) metabolites, with special emphasis on delta 1-tetrahydrocannabinol-7-oic acid (delta 1-THC-7-oic acid), was studied in thirteen heavy Cannabis users after smoking administration of delta 1-THC, followed by a four week discontinuation period. The total amount of delta 1-THC metabolites and the levels of delta 1-THC 7-oic acid could be followed up to 25 days after abstinence using EMIT d.a.u. cannabinoid assay and high-performance liquid chromatography (HPLC). The urinary excretion half-life, calculated from the concentrations of delta 1-THC-7-oic acid versus time, ranged from 0.8-9.8 days with a mean (+/- SD) of 3.0 +/- 2.3 days. Most of the delta 1-THC-7-oic acid was excreted as conjugate and only trace amounts of unconjugated delta 1-THC-7-oic acid were detected. The total concentrations of delta 1-THC-7-oic acid in urine were compared to the concentrations of "cross-reacting cannabinoids", within the linear range of 20-75 ng/mL, obtained in the semiquantitative EMIT d.a.u. cannabinoid assay. The average ratio of "EMIT concentrations"/delta 1-THC-7-oic acid concentrations obtained by HPLC analysis was 1.23 +/- 84% (C.V.) for 78 urine samples. A total of 83% of the samples with positive EMIT levels (cutoff 20 ng/mL) was confirmed by HPLC analysis (cutoff 7 ng/mL). PMID- 2550703 TI - Lack of interference by nabilone in the EMIT d.a.u. cannabinoid assay, Abbott TDx cannabinoid assay, and a sensitive TLC assay for delta 9-THC-carboxylic acid. PMID- 2550704 TI - Characterization of MCF 7 breast cancer cell growth inhibition by the antiestrogen nitromifene (CI 628) and selected metabolites. AB - Besides undergoing O-demethylation in vivo, the triarylethylene antiestrogen nitromifene [1-(4-(2-pyrrolidinylethoxy)phenyl)-1-(4-methoxy)-phenyl-2-phenyl- 2- nitroethene, 1] undergoes biotransformation via nitroreduction, ethene bond cleavage, and pyrrolidine ring oxidation affording ketone metabolites 2 and 3 and a lactam metabolite 4. Estrogen receptor (ER) affinities of 1, 2, and 4 were, in turn, 1.7, 0.1, and 3.8% that of estradiol in MCF 7 human breast cancer cells, and these compounds inhibited by 50% the proliferation of MCF 7 cells at respective concentrations of 1.1, 5.6, and 2.0 microM. The inhibitory effect of 4 was fully reversible by estradiol, but that of 2 was only partially reversible. Also 3, which did not interact with ER, inhibited proliferation by 44% at a concentration of 10 microM. These results suggested that in contrast to 4, the effects of 2 and 3 were due in part to interaction with sites distinct from ER. Antiestrogen binding sites and calmodulin have been suggested to mediate antiproliferative effects of drugs. Interaction of ligands with the former sites has been proposed to antagonize the growth promoting effect of histamine. Although 2 and 3 had high affinities for these sites, their inhibitory effects on MCF 7 cell growth were largely unaffected by the presence of histidine, the source of intracellular histamine. Thus, the relationship between antiestrogen binding site affinity and antiproliferative effects of 2 and 3 was not clarified. In contrast, MCF 7 cell growth suppression potencies paralleled calmodulin antagonist potencies of 1 and 2 suggesting that interaction of 1 and 2 with calmodulin may contribute to their anticancer effects. PMID- 2550705 TI - The influence of heterology, enzyme label and assay conditions on the sensitivity of microtitre plate enzymeimmunoassays for progesterone in milk. AB - Seven antisera raised against 11 alpha-hydroxyprogesterone 11-hemisuccinate (P11 HS) were used in microtitre plate enzymeimmunoassays (EIAs) for progesterone to identify improvements in sensitivity achievable by using various heterologous labels. EIAs using beta-galactosidase linked to P11-HS, 11 alpha hydroxyprogesterone 11-hemimaleate (P11-HM), 11 alpha-hydroxyprogesterone 11 glucuronide (P11-Glu) or progesterone 3-(o-carboxymethyl) oxime (P3-CMO) were compared. Loss of sensitivity through bridge recognition was least evident using the P11-Glu derivative. The same seven antisera were used to evaluate assay sensitivity using beta-galactosidase, alkaline phosphatase, penicillinase and peroxidase linked to P11-HS or P11-Glu as label. Consistent improvements were achieved with the heterologous assays in the order penicillinase greater than alkaline phosphatase/peroxidase greater than beta-galactosidase: with penicillinase, sensitivity generally exceeded that of RIA. These data provide evidence for the general efficacy of the combination 11 alpha-hemisuccinate (immunogen bridge) and 11 alpha-glucuronide (label bridge) in reducing bridge recognition. EIA performed at 4 degrees C provided greater sensitivity than at ambient temperature (21 degrees C) or 40 degrees C, however, ambient temperature incubation provided a practical compromise. Equilibrium was not achieved under any of the conditions investigated. PMID- 2550706 TI - An experimental study of the anti-HSV-II action of 500 herbal drugs. AB - Experimental assessments were made on the anti-HSV-II action of 500 herbs by determinations of the virus inhibition logarithm (VIL). 13 highly effective herbs (VIL greater than or equal to 4.00) were screened cut, providing a rational basis for clinical therapy. Among these effective herbs, 10 were aqueous extracts of Artemisia anomala, Centella asiatica, Epimedium Sagittatum, Hibiscus mutabilis, Hosta plantaginea, Hypericum japonicum, Inula japonica, Mosla punctata, Rhododendron simsii, and Rhus chinenses, while 3 were alcohol extracts of Epimedium Sagittatum, Hypericum japonicum, and Mosla punctata. PMID- 2550707 TI - Two pathways for inducing interleukin 2 production in MOLT 16 cells, a human leukemic T-cell line: interleukin 1 pathway and interleukin 1-independent pathway. AB - Phytohemagglutinin (PHA)-stimulated MOLT 16 cells (a human leukemic T cell line, at a concentration of 2 x 10(6) cells/ml) secreted 6 U/ml of interleukin 2 (IL 2) into the culture supernatant during 24 h culture. When interleukin 1 (IL 1) (5 U/ml) or IL 1-producing cells such as human T cell leukemia virus-1 (HTLV-1) transformed T cell line, C5/MJ, and myelomonocytic cell line, THP-1-O, were added to the MOLT 16 culture at a concentration of 4 x 10(5) cells/ml, far greater IL 2 production (greater than 65 U/ml) was observed. The activity of soluble IL 1 and membrane-associated IL 1 produced by these accessory (A) cells was completely neutralized by the treatment with anti-human IL 1 antibody. In sharp contrast, MOLT 16 cells co-cultured with BALL-1 cells (a human leukemic B cell line) resulted in comparable increases in IL 2 production (175 U/ml), although no IL 1 or IL 1-like activity was detected either in the supernatants or in the cell lysates of BALL-1 cells. An augmentation of IL 2 production was also induced with paraformaldehyde-fixed BALL-1 cells, and this augmentation could not be inhibited by anti-human IL 1 antibody. These data indicate that MOLT 16 provides a model for determining pathways for activating T cells leading to IL 2 production in response to mitogens. Two pathways have been described in the report: one involving secreted or membrane-associated IL 1, and a second independent pathway which involves contact of surface membranes in the presence of PHA. PMID- 2550708 TI - Apparent Epo-independence of erythroid cells infected with the polycythemia inducing strain of Friend spleen focus-forming virus is not due to Epo production or change in number or affinity of Epo receptors. AB - The polycythemia-inducing strain of the Friend spleen focus-forming virus (SFFVP) induces an acute erythroleukemia in mice. Erythroid cells from these mice differ from normal erythroid cells in that they can proliferate and differentiate in the apparent absence of the erythroid hormone erythropoietin (Epo). Although it was recently shown that the unique envelope protein encoded by SFFV is responsible for altering the hormonal requirements of erythroid cells for growth and differentiation, the mechanisms by which this occurs is not known. Since the SFFV envelope protein appears to interact with a target present only in erythroid cells and since Epo is specific for these cells, it is possible that the virus is exerting its effect through this hormone. In an effort to ascertain if this is the case, we examined cells from SFFVP-infected mice to determine (a) if they produce Epo or other erythroid growth factors that stimulate erythroid cells to grow in an autocrine-like manner and (b) if they express elevated numbers of Epo receptors that may result in a reduced requirement for the level of Epo needed for growth and differentiation. Our results indicate that SFFVP-infected cells do not secrete Epo or any other erythroid growth factors that could account for the reduced hormonal requirements of these cells. Also, our studies using iodinated Epo in cell binding assays and cross-linking studies indicate that SFFVP-infected cells are not significantly different from normal erythroid cells in the number, affinity, or size of their Epo receptors. PMID- 2550710 TI - [New types of analyses in clinical cytology improve diagnosis and treatment of tumors]. PMID- 2550709 TI - [The man behind the syndrome: Max Wilms. A famous pathologist and a skillful surgeon]. PMID- 2550711 TI - Cholera toxin enhances ischemia-induced arrhythmias in the isolated rat heart- involvement of a guanine nucleotide binding protein (Gs). AB - Cholera toxin (CTX) at a dose, which disturbed the intestinal functions, was administered into the rat via the tail vein. At 3 hr after injection, the heart was removed and perfused or subject to global ischemia in the Langendorff isolated heart preparation. Electrocardiogram (ECG) was recorded throughout the experiment. The myocardial cAMP content was measured in the intact non-ischemic heart, and in the isolated ischemic heart at 2.5, 5 and 10 min after ischemia. It was found that the incidence and severity of malignant ventricular arrhythmias including ventricular tachycardia (VT) and ventricular fibrillation (VF) was significantly increased during ischemia in the CTX treated group. The cAMP content was also significantly increased in the CTX treated group in both intact non-ischemic and ischemic hearts, indicating an activation of the guanine nucleotide regulatory protein (Gs). The results of the present study provide evidence that activation of Gs during ischemia may also contribute to the genesis of arrhythmia. PMID- 2550712 TI - Purification of bovine angiotensin converting enzyme. AB - A change has been made in the commonly used lisinopril affinity gel procedure for purifying angiotensin converting enzyme. The new method greatly decreases the time required and greatly increases the yield of pure enzyme. All of the enzyme in various bovine tissues was extracted with 0.5% triton X-100 and applied to the affinity column; 70% was trapped and all of the trapped enzyme was released as the apoenzyme by EDTA. The holoenzyme was recovered by dialysis against zinc containing buffer. The turnover numbers were precisely the same for enzyme from lung, atrium, kidney, striatum and blood. The tissue concentrations of ACE were very different but the final specific activities were the same. PMID- 2550713 TI - Behavioral and neurotrophic activity of ACTH-(7-16)NH2. AB - The behavioral and neurotrophic effects of ACTH-(7-16)NH2 were assessed in a number of tests in which other ACTH fragments are active. Subcutaneous injection of ACTH-(7-16)NH2 increased motor activity of group-housed rats tested under low light intensity and induced hypokinesia in rats subjected to the mild stress of a nonfunctional "hot" plate. In rats with 6-OHDA lesions in the nucleus accumbens daily subcutaneous treatment with ACTH-(7-16)NH2 during the first week following the lesions reversed the lesion-induced motor hypoactivity. The ED50's for the effects of ACTH-(7-16)NH2 on the environmentally induced changes in motor activity, the stress-induced hypokinesia and the impaired motor activity of 6 OHDA lesioned rats were approximately 8 micrograms/kg. 6.3 micrograms/kg and 0.45 micrograms/kg respectively. It is concluded that ACTH-(7-16)NH2 may mimic the effect of an ACTH-like peptide in the brain involved in brain processes triggered by changes in the environment and by brain damage. PMID- 2550714 TI - Effects of omega-3 fatty acids on vascular smooth muscle cells: reduction in arachidonic acid incorporation into inositol phospholipids. AB - A rapid increase in arachidonic acid incorporation into phosphatidylinositol (PI) occurred following exposure of cultured porcine pulmonary artery smooth muscle cells to calcium ionophore A23187. This response was specific for PI and phosphatidic acid; none of the other phosphoglycerides showed any increase in arachidonic acid incorporation. The incorporation of [3H]inositol also was increased, indicating that complete synthesis of PI rather than only fatty acylation occurred in response to the ionophore. The presence of omega-3 fatty acids, especially eicosapentaenoic acid (EPA), reduced arachidonic acid but not inositol incorporation into PI. Stimulated incorporation of EPA also occurred under these conditions, suggesting that EPA replaces arachidonic acid in the newly synthesized pool of PI. Although much less arachidonic acid was incorporated into the polyphosphoinositides following exposure to the ionophore, arachidonic acid incorporation into these phosphorylated derivatives also decreased when EPA was present. These findings suggest that when omega-3 fatty acids are available, less arachidonic acid is channeled into the inositol phospholipids of activated smooth muscle cells because of replacement by EPA. This may represent a mechanism whereby omega-3 fatty acids, especially EPA, can accumulate in the metabolically active pools of inositol phospholipids and thereby possibly influence the properties or responsiveness of vascular smooth muscle. PMID- 2550715 TI - Ca2+ transport across the plasma membrane of striated muscle. AB - In both types of striated muscle (skeletal and cardiac), calcium flux across the plasma membrane (sarcolemma) is regulated by at least three distinct membrane proteins; a voltage-dependent Ca2+ channel, Ca2+ pump (Ca2+ ATPase), and the Na+/Ca2+ antiporter. Each of these proteins is subject to regulation by intracellular second messengers. The magnitude and the role of this transsarcolemmal calcium flux are quite different between cardiac and skeletal muscle. In cardiac muscle, the influx is large, precedes, and is obligatory for contraction. There is general agreement that this influx is the trigger for Ca2+ release from the sarcoplasmic reticulum (SR) in the heart according to the Ca2+ induced Ca2+ release hypothesis. Variations in the transsarcolemmal Ca2+ influx have a profound effect on the strength of cardiac contraction, and it appears that this is the primary physiological strategy for regulation of contractility. In skeletal muscle, on the other hand, the T-tubules represent the richest source of dihydropyridine (DHP)-sensitive calcium channels known to exist, yet the influx of Ca2+ is proportionally much smaller and a significant portion enters the fiber following the twitch. While the majority of the Ca2+ influx is twitch dependent, it is quite clear that contraction in skeletal muscle is not predicated on this influx. It has been proposed that these DHP channels act as voltage sensors in order to initiate release of SR Ca2+; however, the link between the sensors and the opening of the SR Ca2+ (ryanodine-sensitive) channel is unknown. Transsarcolemmal Ca2+ transport appears to be subject to intense regulation to modify the acute response and demonstrates some plasticity in the adaptation to chronic perturbations. PMID- 2550716 TI - Imaging tumors with respect to a grid coordinate system by proton magnetic resonance using lipid suppression and T2 weighting. AB - Experimental tumors can be located with respect to a grid coordinate system by using lipid-suppressed T2-weighted proton imaging with thick coronal slices and a plexiglass grid containing a solution of paramagnetic ions. Most normal tissues except the brain, eyes, and CNS are essentially transparent under these conditions, allowing thick sections to be rapidly searched for tumors. Use of this approach to monitor tumor growth is demonstrated. PMID- 2550717 TI - High-flux signals and spatial localization in high-resolution 1H spectroscopy with surface coils. AB - To perform in vivo localized proton spectroscopy with water suppression, spin echo sequences, made of binomial pulses, are commonly used with surface coils. The frequency selective response to such a sequence is also-spatially dependent, that is dependent on the sample shape and on the pulse angle adjustment. It is consequently pointed out in this paper that quantitative analysis for relative peak intensities may be strongly affected by the contribution of the high-flux regions. In vivo proton spectroscopy of rat brain exemplifies this difficulty. It is shown that the use of selective prepulses to suppress high-flux signals may be of poor efficiency depending on chemical shift, while the use of hard nonselective prepulses works for any chemical shift. PMID- 2550718 TI - Study of proton spin-lattice relaxation variation induced by paramagnetic antibodies. AB - A murine anti-human melanoma monoclonal antibody fragment was labeled with gadolinium and its proton relaxation efficiency compared to controls at frequencies ranging from 2 to 300 MHz. Relaxation time variations were about 30 40% in 10-15 microM solutions. The labeled fragment showed proton relaxation enhancement relative to free gadolinium, while preserving its immunoreactivity. A tentative labeling of a melanoma pellet by means of the fragment, just at the borderline of a minimum expected T1 variation, gave no detectable difference. PMID- 2550719 TI - Relative contributions of chemical exchange and other relaxation mechanisms in protein solutions and tissues. AB - Transverse relaxation times T2 of water protons were measured in 5% protein solutions and soaked rat liver in different static magnetic fields (0.15 to 11 T). Protein molecular weight varied between 1.4 and 480 kDa in solutions of varying degrees of deuteration. The data obtained are analyzed in terms of a model system consisting of three phases of different relaxation characteristics: protein protons, hydration layer water protons, and bulk water protons. The contributions to relaxation due to hydrodynamic effects on water protons, cross relaxation between the hydration layer water protons and the protein protons, and chemical exchange between the hydration layer water protons and the bulk water protons are separately estimated. The experimental results indicate that the "hydrodynamic interactions" are about the same magnitude in rat liver and all the proteins studied, but the contribution of the cross relaxation differs by several orders in different protein systems. Fast chemical exchange between the hydration layer water and the bulk water causes considerable shortening of T2 at high magnetic fields for all the protein solutions and rat tissue studied. Selected samples were studied at different temperatures (213-318 K) and with different intervals in the CPMG sequence. The rates of chemical exchange and fractional populations of different phases are determined, and the results obtained provide support for the model in which fast exchange among the different water phases is an important feature of the overall relaxation behavior. PMID- 2550720 TI - On the use of double-quantum coherence from an AX3 system (protons in lactate) for spectral editing. AB - The suppression of water signals in in vivo proton spectra by means of multiple quantum filtering can be brought about in several ways. This communication exposes the shortcomings of using purely subtractive filtering techniques to observe signals from metabolites that contain proton couplings of the form AX3. PMID- 2550721 TI - A dual-tuned probe and multiband receiver front end for X-nucleus spectroscopy with proton scout imaging in vivo. AB - A dual-tuned volume coil probe and a novel multituned receiver front end are described, for spectroscopy in vivo of X nuclei with scout imaging of protons. Detailed circuit information is given for the probe, diplexer, receiver protection switch, and preamplifier. PMID- 2550723 TI - Mutagenicity in V79 Chinese hamster cells of n-alkanals produced by lipid peroxidation. AB - The mutagenicity for mammalian cells of five n-alkanals produced by lipid peroxidation was tested in V79 Chinese hamster lung cells either at the hypoxanthine-guanine phosphoribosyltransferase locus as resistance to 6 thioguanine or at the Na/K ATPase locus as resistance to ouabain. The results show that propanal, butanal, pentanal and hexanal induced a dose-dependent increase in the frequency over controls of both 6-thioguanine- and ouabain resistant mutants at concentrations ranging from 3 to 30 mM. With nonanal the same effects were observed with concentrations of 0.1-0.3 mM. PMID- 2550722 TI - Impaired recovery and mutagenic SOS-like responses in ataxia telangiectasia cells. AB - Radiosensitive fibroblasts from patients with ataxia telangiectasia (AT) were studied for their proficiency in two putative eukaryotic SOS-like responses, namely the enhanced reactivation (ER) and enhanced mutagenesis of damaged viruses infecting pre-irradiated versus mock-treated cells. A previous report indicated that, unlike normal human cells, a line of AT fibroblasts (AT5BIVA) could not be induced to express ER of damaged parvovirus H-1, a single-stranded DNA virus, by UV- or X-irradiation. In the present study, AT5BIVA fibroblasts were also distinguished from normal cells by the inability of the former to achieve enhanced mutagenesis of damaged H-1 virus upon cell UV-irradiation. In contrast, dose-response and time-course experiments revealed normal levels of ER of Herpes simplex virus 1, a double-stranded DNA virus, in X- or UV-irradiated AT5BIVA cells. Taken together, these data point to a possible deficiency of AT cells in a conditioned mutagenic process that contributes to a greater extent to the recovery of damaged single-stranded than double-stranded DNA. Such a defect may concern the replication of damaged DNA or the generation of signals promoting the latter process and may be related to the lack of radiation-induced delay that is typical of AT cell DNA synthesis. PMID- 2550724 TI - H4IIEC3 rat hepatoma cells activate N-nitrosodimethylamine but are resistant to the genotoxic products. AB - Exposure of the differentiated rat hepatoma cells H4IIEC3 (H4) or several of their subclones to 20 mM N-nitrosodimethylamine (NDMA) did not significantly increase the number of 6-thioguanine (TG)-resistant cells or of micronuclei. Similar results were obtained with H4 cells pretreated with isopropanol, an inducer of the NDMA-metabolizing cytochrome P450 form. However, when H4 cells were co-cultured with V79 Chinese hamster cells, which are incapable of activating NDMA, exposure to the nitrosamine (5-40 mM) caused a concentration dependent increase in the frequency of TG-resistant V79 cells; pretreatment of H4 cells with 0.5% isopropanol nearly doubled the magnitude of this response. When freshly isolated rat hepatocytes were used as the activation system in co cultures with either H4 cells or V79 cells, NDMA induced up to 20 times more TG resistant mutants in V79 cells than in H4 cells. The results indicate that H4 hepatoma cells are capable of metabolizing NDMA to genotoxic products but can protect themselves, presumably by repairing the potentially mutagenic DNA lesions. PMID- 2550725 TI - Kinetic properties of Na+/H+ exchange and Li+/Na+, Na+/Na+, and Na+/Li+ exchanges of human red cells. PMID- 2550727 TI - Sodium-calcium and sodium-proton exchangers in red blood cells. PMID- 2550726 TI - Water channels across the red blood cell and other biological membranes. PMID- 2550728 TI - Alkali metal/proton exchange. PMID- 2550729 TI - Na+, K+-pump stoichiometry and coupling in inside-out vesicles from red blood cell membranes. PMID- 2550731 TI - Measurement of amino acid transport by hepatocytes in suspension or monolayer culture. PMID- 2550730 TI - Ca2+ fluxes and phosphoinositides in hepatocytes. PMID- 2550732 TI - Preparation and culture of embryonic and neonatal heart muscle cells: modification of transport activity. PMID- 2550733 TI - Measurement of Na+ pump in isolated cells. PMID- 2550734 TI - Measurement of Na+-K+ pump in muscle. PMID- 2550735 TI - Purification and reconstitution of the phosphate transporter from rat liver mitochondria. PMID- 2550736 TI - Measurements of cytoplasmic pH and cellular volume for detection of Na+/H+ exchange in lymphocytes. PMID- 2550737 TI - Cation fluxes in the red blood cell: Na+,K+ pump. PMID- 2550738 TI - Reconstitution of intracellular vesicle fusion in a cell-free system after receptor-mediated endocytosis. PMID- 2550739 TI - Digitonin permeabilization procedures for the study of endosome acidification and function. PMID- 2550741 TI - A serological survey of swine parvovirus infection in Italy. AB - A serological survey to detect the presence of porcine parvovirus (PPV) infection in Italy and its geographic distribution was conducted. 1,332 samples of serum collected in 1983/1984/1985 were taken from pig breeding herds and, to a lesser extent, from fattening piggeries of representative regions of Italy. They were tested using the hemagglutination inhibition test (HIT). The results of the serological study indicate that parvovirus infection is widespread in Italian herds having 70.3% of sera antibodies to this virus; the most frequent titer detected was 1:8192 or higher. Sera from adult sows showed a higher rate (73.1%) of positive titers than those from 3-6 month-old pigs (40.7%). Positive samples were more relevant in the regions of northern Italy when compared with those from the southern Italy, Sicily and Sardinia. PMID- 2550740 TI - Membrane lipid components interacting with hepatitis A virus. AB - The involvement of lipid components in hepatitis A virus (HAV) attachment to host cells has been investigated. Isolated Frp/3 cell membranes and whole lipids, phospholipid and glycolipid fractions extracted from them were able to bind the virus and to prevent infection. Treatment of virus with various phospholipids or glycolipids demonstrated the participation of phosphatidylserine, phosphatidylethanolamine and galactose in HAV binding. Results obtained added further information on the receptor specificity of host cells for HAV. PMID- 2550742 TI - Modification of norfloxacin inhibition of DNA gyrase induced by a 28 KDal DNA binding protein. AB - We have previously studied a clinical isolate of Providencia stuartii which showed high levels of resistance to 4-quinolones, aminoglycoside and beta-lactam antibiotics (Landini et al., 1987). DNA gyrase from this isolate was inhibited for 50% of activity at a concentration of 15 microM of norfloxacin, which is about 5-fold higher compared to the 50% inhibitory concentration for a standard DNA gyrase. It has been described that 4-quinolone inhibition of DNA gyrase is caused by their binding to DNA and by the distortion induced in DNA tertiary structure, and that affinity binding of 4-quinolones is different for DNAs in different structures. In order to detect whether the interaction between pAT 153 and a protein able to modify DNA tertiary structure could affect norfloxacin inhibitory concentrations for DNA gyrase we purified from the clinical isolate of Providencia stuartii a DNA binding protein of about 28 KDal which induces changes in supercoiling degree of DNA. Assays of DNA gyrase activity were performed on the complex pAT 153-DNA binding protein-norfloxacin. Results showed an increase from 15 microM to 20 microM of 50% norfloxacin inhibitory concentration for DNA gyrase when pAT 153 was complexed with the 28 KDal protein. PMID- 2550743 TI - Characterization of a parvovirus isolated from a pig fetus. AB - A small hemagglutinating virus belonging to the Parvoviridae Family was isolated from a 70 day-old pig fetus in a breeding herd in which infertility, metritis and abortions were reported. The virus, named 85/193L, was isolated either in primary pig kidney (PK) cells or in a continuous cell line of minipig kidney origin (MPK), both cell cultures actively growing. It produced a typical cytopathic effect (CPE) starting from the 3rd passage and intranuclear inclusions surrounded by a halo were observed in stained preparations. The isolate was completely resistant to ether, chloroform and to pH 3; it was not inactivated after treatment at 56 degrees C for 4 h and at 70 degrees C for 2 h, while it was completely inhibited by the 80 degrees C/30 min temperature. It contained deoxyribonucleic acid (DNA). The highest infectious titer was reached at 96 h post infection. The infectivity and the hemagglutinating activity of the isolated strain were both inhibited by the reference immune serum against NADL-2 pig parvovirus. This further confirmed that the 85/193L isolate belongs to the parvovirus genus. PMID- 2550744 TI - Universal precautions prevent transmission of HIV. PMID- 2550745 TI - Frequency of cell transformation by the small DNA tumor viruses: infection of proliferating cells and quiescent cells. AB - Small DNA-containing tumor viruses (simian virus 40, mouse polyoma-virus, and adenoviruses) malignantly transform fibroblasts of the susceptible rodents. Fibroblasts can exist, in vitro and in vivo, in either of the two states: the proliferating state or the quiescent state. In the present study, we examined whether the state of fibroblasts at the time of exposure to these DNA viruses affects the frequency of transformation. Dense-focus formation in monolayer culture of rat 3Y1 fibroblasts was used to quantitate transformation. Results show that the frequency of transformation by simian virus 40 and mouse polyomavirus was reduced when cells were in the proliferating state at the time of virus inoculation as compared to cells in the quiescent state, whereas that by adenovirus type 12 was similar in the two cellular states. The reduction of the frequency of transformation in proliferating cells infected with simian virus 40 was also observed in BALB/c 3T3 mouse cells. Mechanisms underlying the difference between the two cellular states and the difference between the papovavirus and adenovirus in this aspect of transformation remain to be investigated. PMID- 2550747 TI - Regulation of thymidine kinase activity in mouse L cells biochemically transformed by varicella-zoster virus. AB - Regulation of thymidine kinase (TK) activity was examined in L(O)c133 and L(H3) cells carrying varicella-zoster virus-TK gene. TK activity of L(O)c133 cells was similarly high in either medium but that of L(H3) cells was high in HAT medium and low in non-HAT medium. Cell growth was well correlated with TK activities of L(O)c133 and L(H3) cells in medium conditions. Regulation of the TK gene in L cells carrying the VZV-TK gene is discussed. PMID- 2550746 TI - Immunopathology of chronic progressive hepatitis in nude mice infected with low virulent mouse hepatitis virus. AB - Progressive hepatitis in athymic nude (nu/nu) mice due to a low-virulent mouse hepatitis virus, MHV-2 cc, was examined for involvement of immunocytes and serum antibodies. At 3 to 6 weeks postinoculation (p.i.) a considerable number of Mac 1 and asialo GM1-positive cells were accumulated in the affected liver and spleen. There were also some Thy-1-positive cells. Later than 2 weeks p.i., serum IgG and IgM antibodies were detected in parallel with virus-neutralizing activity, while the IgG levels were lower than those of infected euthymic (nu/+) littermates. By transfer of the infected nu/nu mouse serum, the recipient euthymic mice acquired resistance to lethal challenge infection with a virulent virus, MHV-2. PMID- 2550748 TI - Possible metabolic consequences of fermentation in the colon for humans. AB - We postulate that the short chain fatty acids, produced in the large gut by the microbial fermentation of dietary fiber, improve glucose tolerance and inhibit hepatic cholesterol and fibrinogen synthesis, probably by preventing an increase in serum levels of free fatty acids, and by improving insulin sensitivity. Since hypercholesterolemia, hyperfibrinogenemia and glucose intolerance are important risk factors for coronary heart disease, this could serve as a basis for recommendations that Western populations at risk should increase their dietary intake of substrates for short chain fatty acids. PMID- 2550749 TI - Epstein-Barr virus as a cause of autoimmune disease and other medical morbidity in patients with affective disorders. AB - We hypothesize that psychiatric patients suffering from the major affective disorders (depression and manic-depressive illness) may commonly also suffer from a chronic active infection with the Epstein-Barr virus. This infection would be a consequence of the immune dysfunction known to be associated with these disorders of mood. According to this hypothesis, the increased medical morbidity and mortality reported in these psychiatric patients would be attributable in part to diseases in which Epstein-Barr virus is implicated or suspected as a cause. PMID- 2550750 TI - Potential human health risks associated with animal retroviruses: some hypotheses. AB - Several studies have demonstrated the existence of morphological, biological, genetic and antigenic correlations between human and animal retroviruses. Furthermore, some agents of bovine and feline origin can also be considered useful comparative models for the pathogenetic study of human diseases such as leukaemia, acquired immunodeficiency syndrome (AIDS) and multiple sclerosis (MS). In spite of the existence of data suggesting a possible role of some animal retroviruses (e.g. the bovine leukosis virus, BLV) in the etiology and epidemiology of human leukaemia and MS, the true zoonotic potential of such agents remains unknown, but their genetic and antigenic linkages with some human lentiviruses (such as the human immunodeficiency virus-1, HIV 1) can make them hypothetically responsible for a certain number of false-positive serological reactions in man, especially in those subjects who are professionally exposed to animal lentiviruses. In the present paper particular reference is made to this last hypothesis which, if confirmed, would suggest a larger scale-utilization of more specific diagnostic tests for HIV infection in man, such as the immunoblotting techniques. PMID- 2550751 TI - Collagenases from periarticular ligaments and tendon: enzyme levels during the development of joint contracture. AB - Our laboratories have previously demonstrated that normal rabbit periarticular ligaments, anterior cruciate ligaments (ACL), medial collateral ligaments (MCL) and patellar tendon (PT) secrete collagenase. In this current study we examined these connective tissues following an immobilization period of 4 weeks. In the ligaments producing collagenase, activity was expressed only in the control, not in the immobilized joint. Control and experimental patellar tendon samples produce collagenolytic activity, suggesting that the expression of enzyme is less affected in tendons as compared to ligaments. Characterization of these collagenases was carried out using an antiserum directed against rabbit synovial collagenase. We demonstrated that ligament (ACL) and tendon (PT) collagenases cross react with this antibody in a double immunodiffusion assay. Protein blots of PT, ACL and MCL collagenases identified one major species (Mr = 45,000) and a minor species (Mr = 50,000) of immunoreactive proteins in all three connective tissues. Differences between control and experimental enzyme levels appear to be due to less collagenase protein being produced by immobilized ligaments. PMID- 2550752 TI - Collagenase is expressed by rabbit VX2 tumour cells in syngeneic and xenogeneic hosts. AB - Specific antisera for the connective tissue metalloproteinases, collagenase, gelatinase (type IV collagenase) and stromelysin were used to study their respective localizations in both rabbit primary VX2 tumours and in lung metastatic deposits (frozen immediately after excision). Collagenase was found within some cells of the primary tumour and also bound to the extracellular matrix at discrete sites. Previous studies suggest that this matrix staining represents active enzyme. Stromelysin and gelatinase had a more limited distribution, particularly the latter, but both showed cell and matrix staining. In the lung metastases collagenase and stromelysin occurred less frequently, although both cell and matrix staining were observed; gelatinase was not seen. When rabbit VX2 cells were transplanted into nude mice they grew as a discrete nodule. Cells within this nodule stained with the antiserum to collagenase, which recognizes rabbit but not mouse enzyme, and thus demonstrated that cells of tumoural origin synthesize collagenase in vivo. Stromelysin was also co-localized with collagenase in some tumour cells. PMID- 2550753 TI - Platelet interaction with [125I]-labeled collagen type III: requirement of fibrillar structure formation. AB - Interaction of collagen type III (CIII) with washed human platelets was studied using CIII preparation from human placenta. CIII was labeled with [125I] and [125I]-CIII in monomeric and fibrillar form [( 125I]-CIIIm and [125I]-CIIIf respectively) were incubated with platelets at room temperature. Platelet associated and free labels were separated by centrifugation through 20% sucrose. Binding of [125I]-CIIIf was unsaturable, linearly dependent on the concentration of label and represented 28 +/- 3% of the added protein. In comparison with CIIIf, binding of [125I]-CIIIm was minimal and represents only 0.9 +/- 0.2% of the added protein. The binding of [125]-CIIIm was also nonsaturable and linearly depend on the concentration of the labeled protein. Platelet activation neither increases the CIIIf binding, nor stimulates the binding of CIIIm. The binding of [125I]-CIIIf was not inhibited by the excess of the unlabeled CIIIm. The data obtained suggests the absence of high-affinity collagen receptors in platelets and corroborates the hypothesis of multiple low-affinity interactions between collagen fibrils and platelet surface. Binding of CIIIf was very fast--the level of binding reached a plateau within 1 min, and was similar in the presence of Ca2+/Mg2+ and EDTA. Spectrophotometrically undetectable microfibril formation during the lag phase of fibrillogenesis was sufficient for nearly the same as with large fibrils binding of CIII to platelets. Unlike platelets red blood cells (RBC) fail to bind significant amounts of [125I]-CIIIf. PMID- 2550754 TI - Protein disulphide isomerase, a multifunctional endoplasmic reticulum protein. AB - Protein disulphide isomerase (E.C. 5.3.4.1) has been purified, cloned, and sequenced from a variety of vertebrate tissues. The enzyme and its isoforms have been assigned a role in four functional activities: (1) hydroxylation of proline residues in procollagen; (2) disulphide bond oxidation, isomerization, and reduction; (3) the major non-nuclear binding protein of the thyroid hormone 3,3',5-triiodo-L-thyronine; and (4) a component of oligosaccharide transferase. The concentration of the enzyme has been shown to be positively correlated with an endoplasmic reticulum network which is active in secreting disulphide-bonded polypeptides. The enzyme is directed into the endoplasmic reticulum by virtue of a 19 residue N-terminal signal peptide; a four amino acid C-terminal KDEL sequence prevents the enzyme from being secreted. Careful inspection of the sequence data of the isoforms from human tissues reveals a 97% similarity; whereas, analyses of the data from chick tissues reveals only a 80% level of similarity. Chromosomal localizations using human cDNA probes against different human isoforms have assigned the gene(s) to opposite ends of the long arm of chromosome 17. The compiled data suggest the presence of a family of related polypeptides, all of which reside within the lumen of the endoplasmic reticulum. PMID- 2550756 TI - [Similarity of the causative agent of visceral leishmaniasis from Iraq and Leishmania tropica based on the molecular parameters of kinetoplast DNA]. AB - Restriction analysis demonstrated that the cleavage pattern of kinetoplast DNA of visceral leishmaniasis causative agent from Iraq was similar to that of the anthroponotic cutaneous leishmaniasis causative agent Leishmania tropica, and differed from that of the kinetoplast DNA of Leishmania infantum and L. donovani, typical visceral leishmaniasis causative agents. Similarity was established both for maxi- and minicycle molecules of the kinetoplast DNA. Evolutionary relation between L. tropica and the causative agent of visceral leishmaniasis from Iraq is suggested. PMID- 2550755 TI - Immunological diagnosis in viral infections of the central nervous system: course of antibody titres against homo- and heterologous viruses. AB - In clinical cases suspected for viral encephalitis or meningoencephalitis, the estimation of virus-specific antibodies especially in liquor requires high sensitivity as well as specificity. With enzyme immunoassays the sensitivity in detecting antibodies has increased compared to e.g., complement fixation tests. This report concerns the determination of virus-specific antibodies with a commercial enzyme-linked immunosorbent assay (ELISA) in paired liquor/serum samples of four patients with encephalitis or meningoencephalitis. Up to six virus-specific antibodies of the IgG and IgM classes have been determined [herpes simplex virus (HSV), varicella-zoster virus (VZV), cytomegalovirus, mumps virus, measles virus, and rubella virus]. Additionally, serum samples from several patients suffering, or recovered from, diseases caused by HSV and VZV without CNS involvement have been included as controls. The results showed that besides the virus-specific antibody development (IgG and IgM) against the leading virus, i.e., principally concerned in the disease manifestation assumed to be primarily causing the disease, virus-specific antibodies of the IgG and IgM class against a heterologous virus (e.g., VZV) could also be measured with substantial titers. "Cross-reacting" antibodies to both HSV and VZV with the ELISA only appeared and were present in cases where the infection mainly affected the CNS: no such immunological "cross-reactivity" was observed in serum of individuals in "clinically silent" stages of both HSV and VZV infections. The same situation with no measurable "cross-reacting" antibodies was found in cases of acute HSV or VZV diseases where the CNS was not involved. These findings have been discussed with respect to the findings of common antigens, especially between HSV and VZV, and with respect to an unspecific stimulation of immunocompetent cells. PMID- 2550757 TI - Genetic evidence for superoperonal organization of genes for photosynthetic pigments and pigment-binding proteins in Rhodobacter capsulatus. AB - Three adjacent operons, each concerned with photosynthesis in Rhodobacter capsulatus, have been shown by genetic means to be cotranscribable. In the course of describing the characteristics of the bchCA operon, which encodes two enzymes essential for bacteriochlorophyll synthesis, we found that the expression of the bchCA genes is influenced by readthrough from the upstream crtE and crtF genes. The crtE and crtF genes encode enzymes required for carotenoid biosynthesis and function as an operon. Furthermore, the distal structural gene of the bchCA operon, bchA, contains within it both the major oxygen-regulated promotor (Ppuf1) and the constitutive (Ppuf2) promotor for the puf operon. Since these three operons, crtEF, bchCA, and puf, are all transcribed in the same direction, it appears that polymerases traversing the downstream regions may start at any of several promoters. This pattern of transcription, which is unusual among bacteria, demonstrates that the activities of individual operons in a superoperonal cluster may be affected by their positions within the cluster. PMID- 2550758 TI - Nucleotide sequence and regulation of expression of the Aspergillus nidulans gdhA gene encoding NADP dependent glutamate dehydrogenase. AB - The nucleotide sequence of the Aspergillus nidulans gdhA gene encoding NADP linked glutamate dehydrogenase has been determined and Northern blot analysis used to study the regulation of expression of this gene. The gdhA gene is 1485 nucleotides long and, by comparison with the corresponding Neurospora crassa am gene, has two putative introns of 53 nucleotides and a protein encoding region of 1380 nucleotides that codes for an inferred protein of 49.63 kDa which shows regions of homology with glutamate dehydrogenase proteins from a range of organisms. mRNA analysis of wild-type mycelium grown under a variety of conditions shows that: (a) the highest levels are seen with glucose as the carbon source with inorganic nitrogen; and (b) no gdhA mRNA is detectable when cells are transferred to amino acids as sole carbon source, closely matching the observed glutamate dehydrogenase activity levels under identical conditions. The results presented strongly suggest that a good carbon source is a prerequisite for transcription, but the molecular mechanism responsible is unclear. PMID- 2550759 TI - A homozygous S genotype of Brassica oleracea expresses two S-like genes. AB - The sporophytic self-incompatibility system of Brassica species is controlled by a single locus, S. Recognition of self between pollen and stigma is probably mediated by S locus-specific glycoproteins (SLSGs). We describe the isolation, from an S29 homozygote of Brassica oleracea, of two different cDNA clones for transcripts which are equally abundant in stigmas competent for self incompatibility and each of which is homologous to previously reported SLSG sequences. Extensive DNA sequence divergence between the two clones precludes their cross-hybridisation and each acts as a gene-specific probe. All S genotypes appear to have a single copy of each gene but there are significantly different levels of polymorphism associated with each. The clear structural homology between the two indicates a gene duplication involving the S locus and, perhaps, related to the evolution of self-incompatibility. PMID- 2550760 TI - Regulatory elements involved in the tissue-specific expression of the yellow gene of Drosophila. AB - We have assessed the DNA sequence requirements for the correct spatial pattern and phenotypic expression of y in the late embryo/larvae. The wild-type larval phenotype requires both the regions between -294 bp and -92 bp and a portion of the intron; the sequence element(s) located within the intron can act in a position independent manner to effect the wild-type larval phenotype. The larval expression pattern was examined by tissue experiments in situ and by staining germline transformants derived from various y/lacZ fusion constructs. The larval expression of y is restricted to the mouthparts, microsetae and anal plates. While the -495 bp to +194 bp region alone cannot effect a wild-type larval expression pattern, this region in conjunction with the intron appears to be sufficient to drive beta-gal expression in an essentially wild-type pattern. Our data further suggest that the -294 bp to -92 bp region contains elements which specify the larval pattern and that the element(s) in the intron normally act to enhance the level of expression necessary for the wild-type larval phenotype. We also present a phenotypic analysis of the adult cuticle structures of germline transformants derived from a variety of deletion and rearrangement constructs of the y gene. This analysis has revealed several new features associated with the regulation of y expression. PMID- 2550761 TI - Genetic and structural characterization of the avirulence gene avrBs3 from Xanthomonas campestris pv. vesicatoria. AB - The avirulence gene avrBs3 from Xanthomonas campestris pv. vesicatoria was cloned and found to be localized on a self-transmissable plasmid. Genetic analysis of an avrBs3 insertion mutation revealed that avrBs3 constitutes a single locus, specifying the resistant phenotype on pepper plants. Southern blot experiments showed that no DNA sequences homologous to avrBs3 were present in other races of X. c. pv. vesicatoria, which are unable to induce a hypersensitive reaction on ECW-30R. However, the DNA of several different pathovars of X. campestris hybridized to the avrBs3 probe. A deletion analysis defined a region of 3.6-3.7 kb essential for avrBs3 activity. The nucleotide sequence of this region was determined. A 3561 nucleotide open reading frame (ORF1), encoding a 125,000 dalton protein, was found in the 3.7 kb region that was sufficient for avrBs3 activity. A second long ORF (2351 nucleotides) was identified on the other strand. A remarkable feature of both ORFs is the presence of 17 direct repeats of 102 bp which share 91%-100% homology with each other. PMID- 2550762 TI - Efficient transformation of Bacillus thuringiensis and B. cereus via electroporation: transformation of acrystalliferous strains with a cloned delta endotoxin gene. AB - Electroporation was used as a method to transform intact cells of Bacillus thuringiensis and B. cereus. With our optimized method a range of plasmid vectors could be transformed into strains of B. thuringiensis at frequencies of up to 10(7) transformants/micrograms DNA. This high frequency allows cloning experiments to be done directly in B. thuringiensis. A bifunctional vector capable of replicating in Escherichia coli and in Bacillus spp. was constructed. The kurhd1 protoxin gene was cloned into this shuttle vector to produce plasmid pX193, then transformed into B. thuringiensis HD1 cryB and B. cereus 569K. The cloned protoxin gene was expressed in sporulating cultures of both strain HD1 cryB (pX193) and 569K (pXI93), producing crystal protein active in biotests against larvae of Heliothis virescens. This demonstrates the usefulness of the electroporation method for the introduction of cloned toxin genes, in either their native or modified form, into a variety of host strains. PMID- 2550764 TI - Initiation of DNA replication in Escherichia coli after overproduction of the DnaA protein. AB - Flow cytometry was used to study initiation of DNA replication in Escherichia coli K12 after induced expression of a plasmid-borne dnaA+ gene. When the dnaA gene was induced from either the plac or the lambda pL promoter initiation was stimulated, as evidenced by an increase in the number of origins and in DNA content per mass unit. During prolonged growth under inducing conditions the origin and DNA content per mass unit were stabilized at levels significantly higher than those found before induction or in similarly treated control cells. The largest increase was observed when using the stronger promoter lambda pL compared to plac. Synchrony of initiation was reasonably well maintained with elevated DnaA protein concentrations, indicating that simultaneous initiation of all origins was still preferred under these conditions. A reduced rate of replication fork movement was found in the presence of rifampin when the DnaA protein was overproduced. We conclude that increased synthesis levels or increased concentrations of the DnaA protein stimulate initiation of DNA replication. The data suggest that the DnaA protein may be the limiting factor for initiation under normal physiological conditions. PMID- 2550763 TI - The uvp1 gene of plasmid pR cooperates with mucAB genes in the DNA repair process. AB - We show that a DNA fragment that contains the uvp1 gene of the plasmid pR directs the synthesis in Escherichia coli minicells of a protein of apparent molecular weight 20 kDa. Inspection of the nucleotide sequence of the region reveals an open reading frame that has the capacity to encode a protein of 198 amino acids. The uvp1 gene product has been found, in two different systems, to enhance the recombinational activity of E. coli cells. We have also observed a striking similarity to resolvase and invertase proteins. The significance of this finding for the function of the uvp1 gene product requires further investigation. We conclude that the uvp1 gene encodes a 20 kDa protein which appears to be responsible for enhancement of both UV survival and recombinational activity in E. coli. PMID- 2550765 TI - Chromosomal localization and expression of CBS1, a translational activator of cytochrome b in yeast. AB - Translation of mitochondrial cytochrome b RNA in yeast requires the product of the nuclear gene CBS1, a 27.5 kDa soluble mitochondrial protein. In this paper we show that the CBS1 gene is located on chromosome IV immediately adjacent to COX9, the gene coding for cytochrome c oxidase subunit VIIa. CBS1 is transcribed as a very low abundant 900 b RNA. Transcription starts at a single position 101 bp upstream of the CBS1 initiation codon. At positions -39 to -27 of its leader sequence it contains a small open reading frame of 4 codons. By monitoring the beta-galactosidase activity of a CBS1/lacZ fusion construct we show that expression of CBS1 is subjected to regulation by oxygen and by glucose: the beta galactosidase activity is elevated threefold in glycerol or galactose grown cells compared to that in glucose grown cells. A further threefold reduction of the activity is observed in anaerobically grown cells. In accordance with this result is the observation that the steady-state level of CBS1 mRNA of anaerobically grown cells is ninefold lower than that of aerobically cultured cells. PMID- 2550766 TI - Molecular cloning of SNM1, a yeast gene responsible for a specific step in the repair of cross-linked DNA. AB - We have isolated yeast gene SNM1 via complementation of sensitivity towards bi- and tri-functional alkylating agents in haploid and diploid yeast DNA repair deficient snm1-1 mutants. Four independent clones of plasmid DNA containing the SNM1 locus were isolated after transformation with a YEp24-based yeast gene bank. Subcloned SNM1-containing DNA showed (i) complementation of the repair-deficiency phenotype caused by either one of the two different mutant alleles snm1-1 and snm1-2ts; (ii) complementation in haploid and diploid yeast snm1-1 mutants by either single or multiple copies of the SNM1 locus; and (iii) that the SNM1 gene is at most 2.4 kb in size. Expression of SNM1 on the smallest subclone, however, was under the control of the GAL1 promotor. Gene size and direction of transcription was further verified by mutagenesis of SNM1 by Tn10-LUK transposon insertion. Five plasmids containing Tn10-LUK insertions at different sites of the SNM1-containing DNA were able to disrupt the function of genomic SNM1 after gene transplacement. Correct integration of the disrupted SNM1::Tn10-LUK at the genomic site of SNM1 was verified via tetrad analysis of the sporulated diploid obtained after mating of the SNM1::Tn10-LUK transformant to a haploid strain containing the URA3 SNM1 wild-type alleles. The size of the poly(A)+ RNA transcript of the SNM1 gene is 1.1 kb as determined by Northern analysis. PMID- 2550767 TI - Cloning and comparison of A alpha mating-type alleles of the Basidiomycete Schizophyllum commune. AB - An A alpha mating-type allele (A alpha 4) was isolated by walking the chromosome from the closely linked PAB1 gene. A cosmid clone containing the A alpha 4 allele isolated from the walk was used as a probe to recover the A alpha 1 allele from another cosmid library. Cosmids encoding mating-type activity were identified by transforming Schizophyllum cells and screening for activation of A-regulated development. Putative mating-type transformants were confirmed in mating tests and genetic analyses of progeny. The identity of the specific alleles isolated was demonstrated by showing that their effectiveness in transforming for mating type is limited to recipient strains possessing an A alpha allele different from the one encoded by the cloned sequences. Transforming DNA is active in trans, suggesting that A alpha encodes a diffusible product. Restriction mapping shows that A alpha 1 and A alpha 4 are coded in the same physical region of the genome, but within a subregion that contains extensive sequence divergence. In addition, Southern analyses show that there is only one copy of A alpha 1 or A alpha 4 per haploid genome, and that they do not cross-hybridize to one another or to any of the other A alpha alleles. A alpha 1 and A alpha 4 were subcloned as 2.8 and 1.2 kb fragments, respectively, retaining in transformation all the mating-type activity demonstrated of the original cosmids. PMID- 2550768 TI - Instability in the ctMR2 strain of Drosophila melanogaster: role of P element functions and structure of revertants. AB - Simultaneous multiple transpositions and long-term genetic instability have been described in the ctMR2 strain of Drosophila melanogaster and its derivatives. This strain originated from a cross that was dysgenic in the P-M system. While spontaneous instability declined over 2 years, instability has been reactivated by backcross to the progenitor P element bearing strain MRh12/Cy. We show here using germline transformation that active P factor alone cannot mimic the effect of this cross, suggesting that MRh12/Cy contains some other activator. In addition, we have observed that ct+ exceptional progeny arise in the F1 as well as the F2 generations. Molecular analysis of X chromosomes from some ct+ progeny indicates that phenotypic reversion of the ct mutation can arise through two unrelated mechanisms. PMID- 2550769 TI - I elements of Drosophila melanogaster generate specific chromosomal rearrangements during transposition. AB - We report a detailed molecular analysis of three chromosomal rearrangements, which have been produced during I-R hybrid dysgenesis in Drosophila melanogaster. They all disrupt the yellow gene. One of them is a deletion; the other two are inversions, which may be interpreted as the results of recombination events between I elements inserted at their break points. These events appear to occur at the time of transposition and involve integrating rather than resident I elements. They are produced by a mechanism very similar to homologous ectopic recombination. PMID- 2550771 TI - Effect of base pair mismatches on recombination via the RecBCD pathway. AB - The effect of base pair mismatches on recombination via the RecBCD pathway was studied in mutS and wild-type Escherichia coli, using substrates that contain single or multiple mismatches. Recombination between homologous DNA inserts in lambda phage and pBR322-derived plasmids forms phage-plasmid cointegrates that result from an odd numbers of crossovers. In the mutS host, when the sequence homology of a pair of 405 bp substrates decreased from 100% to 89%, the recombinant frequency decreased by about 9-fold, while in the wild-type host the decrease was about 240-fold. These results suggest that multiple mismatches can reduce recombinant frequencies by impeding the mechanism of recombination itself, and by provoking mismatch repair. Single mismatches in 31 bp substrates caused reductions in recombinant frequencies of 2- or 12-fold, depending on the location of the mismatch. However, unlike the reduction by multiple mismatches, the reduction of the recombinant frequencies by single mismatches was the same in both mutS and wild-type hosts. Thus a single mismatch is sufficient to impede recombination, and mismatch repair seems unable to act on single mismatches in very short homologies during recombination. PMID- 2550770 TI - Expression and DNA sequence of RED1, a gene required for meiosis I chromosome segregation in yeast. AB - Genetic studies have previously demonstrated that the RED1 gene of Saccharomyces cerevisiae is required for chromosome segregation at the first meiotic division. Northern blot hybridization analysis indicates that the RED1 gene produces two transcripts of 2.75 and 3.2 kilobases. The major 2.75 kb transcript is not present in mitotic cells and is meiotically induced to accumulate maximally just prior to the meiosis I division. The DNA sequence of the RED1 gene was determined and used to predict the amino acid sequence of the encoded gene product. The RED1 protein is 827 amino acids in length and has a molecular weight of 95.5 kilodaltons. There is no significant homology between the RED1 amino acid sequence and other known protein sequences, including those encoded by genes essential for meiosis. PMID- 2550773 TI - Identification of a specific site required for rapid heterologous desensitization of the beta-adrenergic receptor by cAMP-dependent protein kinase. AB - The molecular basis for heterologous desensitization of the beta-adrenergic receptor (beta AR) was investigated by site-directed mutagenesis of the beta AR protein. Rapid heterologous desensitization of agonist-stimulated adenylyl cyclase activity was observed when L cells expressing the wild-type beta AR were incubated with 50 nM epinephrine. This desensitization response could be mimicked in a cell-free system by incubation with cAMP-dependent protein kinase (cA.PK). Deletion of amino acid residues 259-262 from the beta AR, removing one of the two consensus sequences in the receptor for phosphorylation by cA.PK, abolished the ability of the receptor to undergo rapid heterologous desensitization. In contrast, deletion of the other cA.PK consensus sequence (residues 343-348) or truncation of the Ser/Thr-rich C-terminal tail of the beta AR (deletion of residues 354-418) did not affect this heterologous desensitization process. These results suggest that the action of cA.PK on amino acid residue(s) contained within the sequence 259-262 of the beta AR is required for rapid heterologous desensitization of the receptor in response to agonists. PMID- 2550772 TI - Mapping of chromosomal loci associated with lipopolysaccharide synthesis and serotype specificity in Vibrio cholerae 01 by transposon mutagenesis using Tn5 and Tn2680. AB - Vibrio cholerae strains of the 01 serotype have been classified into three subclasses, Ogawa, Inaba and Hikojima, which are associated with the O-antigen of the lipopolysaccharide (LPS). The DNA encoding the biosynthesis of the O-antigen, the rfb locus, has been cloned and analysed (Manning et al. 1986; Ward et al. 1987). Transposon mutagenesis of the Inaba and Ogawa strains of V. cholerae, using Tn5 or Tn2680 allowed the isolation of a series of independent mutants in each of these serotypes. Some of the insertions were mapped to the rfb region by Southern hybridization using the cloned rfb DNA as a probe, confirming this location to be responsible for both O-antigen production and serotype specificity. The other insertions allowed a second region to be identified which is involved in V. cholerae LPS biosynthesis. PMID- 2550774 TI - Topoisomerase II-mediated DNA cleavage by amonafide and its structural analogs. AB - Treatment of SV40-infected monkey cells with amonafide (benzisoquinolinedione), an intercalative antitumor drug, resulted in rapid accumulation of linearized intracellular SV40 DNA molecules that were protein linked. Studies using purified mammalian DNA topoisomerase II have shown that amonafide and its structural analogs interfere with the breakage-rejoining reaction of the enzyme by stabilizing a reversible enzyme-DNA "cleavable complex." Denaturation of the cleavable complex with sodium dodecyl sulfate resulted in DNA cleavage and the covalent association of topoisomerase II polypeptides with the cleaved DNA. Unwinding measurements indicate that amonafide is a DNA intercalator. These results suggest that amonafide and its structural analogs (e.g., mitonafide) represent a new class of intercalative topoisomerase II-active antitumor drugs. Different from other topoisomerase II-active antitumor drugs, amonafide and mitonafide induce specific DNA cleavage at a single major site on pBR322 DNA. The strong site specificity of amonafide may allow detailed characterization of the intercalator-stabilized, topoisomerase II-DNA cleavable complex. PMID- 2550775 TI - Inositol tetrakisphosphate mobilizes calcium from cerebellum microsomes. AB - Ca2+ accumulated by rat cerebellum microsomes in the presence of MgATP was released by added inositol tetrakisphosphate [Ins(1,3,4,5)P4]. The concentrations of D-myo-inositol-1,4,5-trisphosphate [D-Ins(1,4,5)P3], D-Ins(1,3,4,5)P4, and DL Ins(1,3,4,5)P4 required for half-maximal release were 0.15, 4.6, and 7.5 microM, respectively. Maximal concentrations of InsP4 released only 70% of the Ca2+ released by maximal concentrations of Ins(1,4,5)P3. Inositol pentakisphosphate and D-myo-inositol-1,3,4-trisphosphate were relatively inactive. Additional Ca2+ was released when Ins(1,4,5)P3 (or a nonhydrolyzable analog) was added after completion of InsP4-mediated Ca2+ release but not when this sequence of additions was reversed. This indicates that InsP4 releases Ca2+ from part of the InsP3 releasable compartment. No evidence for synergism between InsP4 and InsP3 was obtained and responses to suboptimal concentrations of both inositol phosphates were approximately additive. Heparin was a potent inhibitor of InsP4-mediated Ca2+ release. Inhibition by heparin was competitive with respect to InsP4 concentration and the Ki for heparin was 0.6 microgram/ml (approximately 100 nM), irrespective of whether InsP4 or InsP3 was used as an agonist. A 3-phosphatase capable of converting [3H]Ins(1,3,4,5)P3 to [3H]Ins(1,4,5)P3 could not be detected in cerebellum microsomes. Hence, we conclude that, in vitro, receptors capable of recognizing Ins(1,3,4,5)P4 can also be coupled directly to a Ca2+ release system. PMID- 2550776 TI - Selectivity of quinoxalines and kynurenines as antagonists of the glycine site on N-methyl-D-aspartate receptors. AB - Xenopus oocytes injected with rat brain mRNA were used to identify and characterize the effects of compounds that are antagonists at both the glycine site on N-methyl-D-aspartate (NMDA) receptors and the quisqualate/kainate receptor. Oocytes were voltage-clamped at -60 mV and inward currents were measured at equilibrium following perfusion with agonists and antagonists. Application of 7-chlorokynurenic acid (7-Cl-Kyn) or 6,7-dichloro-3-hydroxy-2 quinoxaline carboxylic acid (6,7-diCl-HQC), each at 15 microM, produced a parallel shift to the right of the glycine concentration-response curve. Schild analysis indicated a KB of 300 nM for 6,7-diCl-HQC and 350 nM for 7-Cl-Kyn. The slopes of the Schild plots were 1.01-1.02 in each case, suggesting that both compounds are competitive glycine antagonists. Both compounds also blocked the receptor mediating kainate-induced inward current. Schild analysis of 6,7-diCl HQC (KB = 3.0 microM, slope = 0.98) indicated competitive antagonism of kainate currents, but with a potency 10-fold lower than at the glycine site. 7-Cl-Kyn antagonized kainate-evoked currents (KB = 14.1 microM), but the slope of the Schild regression was less than 1 (0.72 +/- 0.11; p less than 0.05). Thus, 7-Cl kyn was approximately 40-fold more potent at the glycine site than at the receptor mediating kainate currents but is probably not entirely competitive at the latter receptor. Omission of the Cl groups from these antagonists drastically reduced activity at both glycine and kainate sites. 6,7-Dinitro- and 6-cyano-7 nitro-quinoxalinedione were both more potent antagonists of kainate than glycine, but substitution of Cl at the 6-position and especially the 6- and 7-positions increased potency at the glycine site. These results suggest that the glycine coagonist site of the NMDA receptor and the agonist binding site of the quisqualate/kainate receptor have some structural similarity. Halogenated derivatives of quinoxalines and kynurenines should be useful in evaluating the function of the glycine site in synaptic transmission mediated by NMDA receptors. In this regard we found that 7-Cl-kyn (5 and 15 microM) selectively attenuated NMDA receptor-mediated epileptiform bursts in the CA1 region of hippocampal slices perfused with zero-Mg medium, without reducing the amplitude of the primary population spike. This block could be overcome by 300 microM D-serine, which alone did not influence bursting. These results together indicate that the glycine site plays a role in epileptiform bursting mediated by NMDA receptors in adult rat hippocampus. PMID- 2550778 TI - Fractional vesamicol receptor occupancy and acetylcholine active transport inhibition in synaptic vesicles. AB - Vesamicol [(-)-(trans)-2-(4-phenylpiperidino)cyclohexanol] receptor binding and inhibition of acetylcholine (AcCh) active transport by cholinergic synaptic vesicles that were isolated from Torpedo electric organ were studied for 23 vesamicol enantiomers, analogues, and other drugs. Use of trace [3H]vesamicol and [14C]AcCh allowed simultaneous determination of the concentrations of enantiomer, analogue, or drug required to half-saturate the vesamicol receptor (Ki) and to half-inhibit transport (IC50), respectively. Throughout a wide range of potencies for different compounds, the Ki/IC50 ratios varied from 1.5 to 24. Compounds representative of the diverse structures studied, namely deoxyvesamicol, chloroquine, and levorphanol, were competitive inhibitors of vesamicol binding. It is concluded that many drugs can bind to the vesamicol receptor and binding to only a small fraction of the receptors can result in AcCh active transport inhibition. Possible mechanisms for this effect are discussed. PMID- 2550777 TI - Evidence for the role of epinephrine binding frequency in activation of adenylate cyclase. AB - The binding of epinephrine to beta-adrenergic receptors is a rapid-on, rapid-off process, such that at any level of receptor occupancy (defined as the fraction of time a receptor is bound or, alternatively, the probability that any particular receptor is bound at any given instant) the entire population of available receptors has periods of occupancy that occur at high frequency. While in the bound state, the receptor acts as a mobile catalyst for the activation of adenylate cyclase. Two processes, then, could conceivably contribute to the access of epinephrine-bound receptors to cyclase and the extent of cyclase activation for a given concentration of epinephrine: 1) the rapid switching of epinephrine among receptors ensures that discontinuous distributed regions of the cell surface experience agonist activity and 2) the mobility of the receptors (and GTP-binding protein) in the cell membrane makes it possible for one receptor to activate numerous GTP-binding protein-adenylate cyclase complexes. In principle, either effect can lead to a wide separation between the binding and response curves (EC50 much less than Kd). It has so far been assumed that mobility is able to account completely for the separation. The extent of the contribution of the process of agonist binding and unbinding to adenylate cyclase activation has not been demonstrated or quantified. Here we examine the distinction between binding frequency and receptor mobility contributions to adenylate cyclase activation in epinephrine-stimulated S49 lymphoma cells for which there is a 200-fold separation between the EC50 and Kd at 37 degrees (EC50 = 10 nM, Kd = 2 microM). Experiments were designed to measure adenylate cyclase activation rates for a constant concentration of epinephrine-bound receptors but with variation of the absolute number of receptors involved in the activation. This was accomplished by blocking a portion of the receptor population with an antagonist (propranolol) that has a long occupancy half-life, while increasing the occupancy of the remaining receptors by compensating increases in epinephrine. With this protocol, a condition is approached in which receptor mobility alone is responsible for activation. This resulted in a 50% decrease in adenylate cyclase activity, compared with a control of 30 nM epinephrine. Thus, for epinephrine concentrations near the EC50, the switching of epinephrine among the receptor population is necessary for greater than 50% of the observed activity; it can be shown in conjunction that receptor mobility nonetheless accounts for the majority of the separation between the EC50 and the Kd.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2550779 TI - Growth of S49 cells in low concentrations of beta-adrenergic agonists causes desensitization. AB - Epinephrine at concentrations approximating circulating levels in resting subjects produced significant desensitization in wild type S49 lymphoma cells after long term treatment. Desensitization by such low levels of catecholamines was measured by examining subsequent responses of the cells to higher agonist concentrations and was quantified by comparing the integral cAMP accumulations with time in naive and epinephrine-treated cells challenged with the higher epinephrine concentrations. The cells were significantly desensitized after 8 hr of treatment with 3 nM epinephrine or 3 nM terbutaline and were essentially maximally refractory after 24 hr. The 3 nM epinephrine treatment resulted in a small right shift of the EC50. Responses to epinephrine were partially restored by incubating desensitized cells for 8 hr or longer in growth medium that was free of epinephrine. The attenuation of cAMP responses was largely specific, in that the decrease in the response to prostaglandin was small and the response to forskolin was unchanged. This, together with small increases in cAMP destruction in cell-free preparations from treated cells, suggested that higher phosphodiesterase activity contributed in a minor way to the desensitization. However, the response of the adenylate cyclase system to epinephrine was dramatically attenuated, and very significant changes in the properties of the beta-adrenergic receptors were also obvious. That is, the number of binding sites for epinephrine was reduced by about 65% while the number of sites for [125I]iodocyanopindolol was unchanged. The affinity for the radioactive ligand was significantly reduced. Wild type S49 cells remained viable after several days of continuous treatment with 3 nM epinephrine or terbutaline but responded to subsequent increases in cellular cAMP levels with the expected growth arrest and cytolysis. Involvement of cAMP-dependent protein kinase in this type of desensitization was suggested by the observation that S49 kincells were not desensitized by long term incubation with 3 nM epinephrine. Further, low concentrations of dibutyryl cAMP mimicked the effect of low level epinephrine treatment. We conclude that circulating levels of epinephrine in intact animals are sufficiently high to cause desensitization in cells with sensitivities to the catecholamines in the same range as that of the S49 lymphoma cell in vitro. We would predict that cells with those characteristics would always be at least partially desensitized in vivo. PMID- 2550780 TI - An agonist that is selective for adenylate cyclase-coupled muscarinic receptors. AB - Compound BM5 [N-methyl-N(1-methyl-4-pyrrolidino-2-butynyl) acetamide] has previously been described as an agonist at postsynaptic muscarinic receptors and as an antagonist at presynaptic receptors. In the current work, we studied the ability of this compound to selectively stimulate phosphoinositide (PI) turnover in Chinese hamster ovary cells transfected with m1 muscarinic receptors and in SK N-SH neuroblastoma cells that express only m3 receptors. We also studied the ability of this compound to stimulate adenylate cyclase inhibition in m2 muscarinic receptors from heart tissue and in m4 receptors expressed in NG108-15 cells. BM5 stimulated the two muscarinic receptor subtypes coupled to adenylate cyclase inhibition. In NG108-15 cells, 100 microM BM5 inhibited prostaglandin E1 stimulated cAMP formation by 36 +/- 1.5%, whereas 100 microM of the full agonist oxotremorine-M inhibited cAMP formation by 64.1 +/- 1.9%. The half-maximal concentration for BM5 inhibition of cAMP formation was 0.4 +/- 0.1 microM. In heart membranes, BM5 inhibited isoproterenol-stimulated adenylate cyclase by 24 +/- 2%, whereas oxotremorine inhibited this activity by 34 +/- 3%. In contrast to its activity at these receptor subtypes, BM5 did not stimulate the m1 or m3 receptor subtypes, which couple to PI turnover. In these latter two subtypes, BM5 inhibited oxotremorine-M-stimulated PI turnover with IC50 values of 10-20 microM. Therefore, BM5 is a partial agonist at adenylate cyclase-coupled muscarinic receptor subtypes and is a pure antagonist at PI turnover-coupled muscarinic receptor subtypes. These studies also suggest that, at least in some parts of the brain, postsynaptic muscarinic receptors are coupled to adenylate cyclase, whereas presynaptic muscarinic receptors are coupled to PI turnover. PMID- 2550781 TI - Lactosylated low density lipoprotein: a potential carrier for the site-specific delivery of drugs to Kupffer cells. AB - Low density lipoprotein (LDL) is a spherical particle with a diameter of 22 nm. It consists of an apolipoprotein and a lipid moiety, in which a variety of lipophilic drugs and prodrugs can be incorporated. In the present study, lactose was coupled to the apolipoprotein of LDL by reductive amination (398 +/- 40 residues/LDL particle). After injection into rats, radioactively labeled lactosylated LDL was cleared rapidly from the plasma (half-life, less than 2 min). Ten minutes after injection, the liver contained about 90% of the dose, whereas only small amounts of radioactivity were found in other tissues. Preinjection of N-acetylgalactosamine completely blocked liver uptake, whereas N acetylglucosamine was ineffective. This indicates that the hepatic recognition site is galactose specific. Subcellular fractionation of liver indicated that the recognition of lactosylated LDL is followed by internalization and degradation of the apolipoprotein in the lysosomes. In the liver, Kupffer cells are mainly responsible for uptake. At 10 min after injection, these cells contained a 70 and 7 times higher amount of lactosylated LDL per mg of cell protein than parenchymal and endothelial cells, respectively. After galactose-specific uptake in parenchymal cells was blocked with asialofetuin, the relative concentration in Kupffer cells was even higher. The hepatic uptake of the lipid moiety of lactosylated LDL, labeled with [3H]cholesteryl oleoyl ether, was identical to that of the 125I-labeled apolipoporotein, which indicates that the particle is taken up as a unit. Thus, lactosylated LDL is taken up rapidly and selectively by Kupffer cells, and it appears that it might be a very effective vehicle for the specific delivery of lipophilic drugs, e.g., immunomodulators, to these cells. PMID- 2550782 TI - Transmembrane transport of fatty acids in the heart. AB - Although fatty acid uptake by the myocardium is rapid and efficient, the mechanism of their transmembrane transport has been unclear. Fatty acids are presented to the plasma membrane of cardiomyocytes as albumin complexes within the plasma. Since albumin is not taken up by the cells, it was postulated that specific high affinity binding sites at the sarcolemma may mediate the dissociation of fatty acids from the albumin molecules, before they are transported into the cells. In studies with a representative long-chain fatty acid, oleate, it was in fact shown that fatty acids bind with high affinity to isolated plasma membranes of rat heart myocytes revealing a KD of 42 nM. Moreover, a specific membrane fatty acid-binding protein (MFABP) was isolated from these membranes. It had a molecular weight of 40 kD, an isoelectric point of 9.0, and lacked carbohydrate or lipid components. Binding to a specific membrane protein might represent the first step of a carrier mediated uptake process. Therefore, the uptake kinetics of oleate by isolated rat heart myocytes was determined under conditions where only cellular influx and not metabolism occurred. Uptake revealed saturation kinetics and was temperature dependent which were considered as specific criteria for a facilitated transport mechanism. For evaluation whether uptake is mediated by MFABP, the effect of a monospecific antibody to this protein on cellular influx of oleate was examined. Inhibition of uptake of fatty acids but not of glucose by the antibody to MFABP indicated the physiologic significance of this protein as transmembrane carrier in the cellular uptake process of fatty acids.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550784 TI - Alpha-1-adrenergic stimulation of phosphoinositide breakdown in cultured neonatal rat ventricular myocytes. AB - The regulation of and the intracellular events following alpha 1-adrenergic receptor stimulation in the myocardium still remain to be disclosed. The effect of alpha 1-adrenergic stimulation on phosphoinositide breakdown was studied in cultured neonatal rat ventricular myocytes. Phenylephrine (30 microM) stimulated inositolphosphates formation, but only in the presence of 10 mM LiCl this could be measured. The increase was antagonized by prazosin (1 microM) but not by propranolol (1 microM). The variability in proportional distribution of the three inositolphosphates is discussed. PMID- 2550783 TI - Is there evidence of a role of the phosphoinositol-cycle in the myocardium? AB - The recent findings on a more general involvement of phospholipids in signal transduction and on the different roles of inositolphospholipids in particular, thoroughly complicate research in this field. It becomes increasingly evident that measuring [3H]inositolphosphate formation alone will never provide insight into the complex machinery of cellular signalling. Certainly for the heart in which the role(s) of the inositolphospholipids is far from clarified, the novel trends provide new directions for research. PMID- 2550785 TI - Relationship of byssinosis to the generation of oxygen radicals by bract tissues of cotton plants. AB - Byssinosis is a hazardous respiratory disorder of workers in natural fiber processing industries and, in the case of cotton, is caused by exposure to respirable dust generated from leafy trash associated with raw fibers. To understand the chemical characteristics of involucral trash components that might contribute to bysinosis, we examined the human airway constricting activity and oxygen radical generating activity of dry, frost-killed cotton bracts. In response to inhalation of aerosolized bract extracts, the expiratory flow rates of human volunteers at 40% of vital capacity during partial forced expiration decreased by 3 to 32%. These values enabled us to identify two potentially byssinogenically active bract specimens, a specimen virtually inactive, and a fourth intermediately so. Using spin trapping techniques of electron spin resonance spectrometry, we found that all specimens catalyzed the generation of hydroxyl (preponderantly) and superoxide radicals from hydrogen peroxide. However, the weakest constrictor was the most potent catalyst, and vice versa. This was consistent with transition metal content of the specimens; the most potent catalyst also contained the largest amounts of those metals, suggesting a Fenton-type reaction mechanisms. Other possibilities for the inverse relationship of airway constricting (byssinogenic) activity with oxygen radical generation are discussed. We also found that neither aflatoxin nor endotoxin, contingent contaminants of bracts, catalyzed oxygen radical production from hydrogen peroxide. PMID- 2550786 TI - Adenosine regulates the release of adrenocorticotropic hormone (ACTH) from cultured anterior pituitary cells. AB - We have recently shown the presence of adenosine receptors coupled to adenylate cyclase in anterior pituitary and in the present studies we have investigated the effects of adenosine on ACTH release. The 'R'-site specific analogs of adenosine such as N-Ethylcarboxamide adenosine (NECA), L-N6-phenylisopropyl adenosine (PIA), 2-chloro-adenosine (2-Cl-Ado) all stimulated ACTH release in a dose dependent manner. NECA was the most potent analog and stimulated ACTH release by about 170% with an apparent Ka of 0.1 microM, whereas PIA and 2-Cl-Ado were less potent and stimulated the release by about 110% and 125% with an apparent Ka of 0.2 and 0.4 microM respectively. The stimulation of ACTH release by NECA was inhibited by 3-isobutyl-1-methylxanthine (IBMX). On the other hand, adenosine deaminase (ADA) treatment of the cells also stimulated ACTH release as well as adenylate cyclase activity by about 2-fold, suggesting that endogenous adenosine plays an inhibitory role in the release of ACTH. Other agents, such as corticotropin-releasing factor (CRF), vasoactive intestinal peptide (VIP) and forskolin (FSK) also stimulated ACTH release from these cells. In addition, the stimulation by an optimal concentration of NECA was almost additive with maximal stimulation caused by VIP and FSK. These data suggest that adenosine modulates ACTH release from anterior pituitary through its interaction with adenosine receptors coupled to adenylate cyclase. PMID- 2550787 TI - Hominoid triosephosphate isomerase: regulation of expression of the proliferation specific isozyme. AB - Three primary isoforms of the dimeric glycolytic enzyme, triosephosphate isomerase (TPI; EC 5.3.1.1), are detected in proliferating human cells. The electrophoretically separable isoforms result from the three possible combinations of constitutive subunits and subunits expressed only in proliferating cells. Only a single primary isoform is observed in quiescent cells. The two subunits, which differ by covalent modification (s), are product of the single structural locus for this enzyme. Expression of the proliferation specific subunit (TPI-2) is detected within 6-10 hr following mitogen stimulation of quiescent human cells, requires RNA synthesis and is inhibited by agents which inhibit interleukin 2 expression or function. Only the constitutive subunit (TPI 1) is detected in proliferating cells from nonhominoid primate species. A single class of TPI mRNA, which is increased greater than 10 fold following stimulation of quiescent cells, is detected on northern blot analysis and S1 nuclease digestion analysis of RNA from quiescent and proliferating human cells. It is similar in size to the TPI mRNA from proliferating cells of the African green monkey, a primate species not expressing TPI-2. Comparison of the structure of the TPI gene from rhesus monkey (nonexpressing species) to the gene from expressing species does not suggest a mechanism for generating TPI-2. Thus, the regulation of the expression of the hominoid restricted, proliferation specific subunit of TPI has been further defined, although the mechanism for generating TPI-2 remains elusive. PMID- 2550788 TI - Binding of nuclear factor EF-C to a functional domain of the hepatitis B virus enhancer region. AB - Nuclear factor EF-C is present in extracts prepared from human HepG2 liver cells and from other, nonliver cell lines and binds to the hepatitis B virus and polyomavirus transcriptional enhancer regions in vitro. An inverted repeat (5' GTTGCNNNGCAAC-3') is located within both binding regions. Diethyl pyrocarbonate interference binding assays and competition binding experiments using altered binding sites demonstrated that EF-C contacts symmetrical nucleotides within the inverted repeat. Mutations that changed the length of the spacer region between the arms of the inverted repeat were introduced in the hepatitis enhancer region. Introduction of 1 or 2 base pairs between the repeats did not affect EF-C binding, but deletion of 1 base pair or introduction of 3 to 9 base pairs reduced binding dramatically. Introduction of 10 base pairs restored partial EF-C binding ability. These and other results suggest that EF-C binding is stabilized by dimerization. In vivo assays for enhancer function using these mutants demonstrated that the EF-C binding site is a functional and important component of the hepatitis B virus enhancer region. PMID- 2550789 TI - Platelet-derived growth factor induces rapid and sustained tyrosine phosphorylation of phospholipase C-gamma in quiescent BALB/c 3T3 cells. AB - Platelet-derived growth factor (PDGF) stimulates the proliferation of quiescent fibroblasts through a series of events initiated by activation of tyrosine kinase activity of the PDGF receptor at the cell surface. Physiologically significant substrates for this or other growth factor receptor or oncogene tyrosine kinases have been difficult to identify. Phospholipase C (PLC), a key enzyme of the phosphoinositide pathway, is believed to be an important site for hormonal regulation of the hydrolysis of phosphatidylinositol 4,5-bisphosphate, which produces the intracellular second-messenger molecules inositol 1,4,5 trisphosphate and 1,2-diacylglycerol. Treatment of BALB/c 3T3 cells with PDGF led to a rapid (within 1 min) and significant (greater than 50-fold) increase in PLC activity, as detected in eluates of proteins from a phosphotyrosine immunoaffinity matrix. This PDGF-stimulated increase in phosphotyrosine immunopurified PLC activity occurred for up to 12 h after addition of growth factor to quiescent cells. Interestingly, the PDGF stimulation occurred at 3 as well as 37 degrees C and in the absence or presence of extracellular Ca2+. Immunoprecipitation of cellular proteins with monoclonal antibodies specific for three distinct cytosolic PLC isozymes demonstrated the presence of a 145 kilodalton isozyme, PLC-gamma (formerly PLC-II), in BALB/c 3T3 cells. Furthermore, these immunoprecipitation studies showed that PLC-gamma is rapidly phosphorylated on tyrosine residues after PDGF stimulation. The results suggest that mitogenic signaling by PDGF is coincident with tyrosine phosphorylation of PLC-gamma. PMID- 2550790 TI - Interaction between transcriptional activator protein LAC9 and negative regulatory protein GAL80. AB - In Saccharomyces cerevisiae, transcriptional activation mediated by the GAL4 regulatory protein is repressed in the absence of galactose by the binding of the GAL80 protein, an interaction that requires the carboxy-terminal 28 amino acids of GAL4. The homolog of GAL4 from Kluyveromyces lactis, LAC9, activates transcription in S. cerevisiae and is highly similar to GAL4 in its carboxyl terminus but is not repressed by wild-type levels of GAL80 protein. Here we show that GAL80 does repress LAC9-activated transcription in S. cerevisiae if overproduced. We sought to determine the molecular basis for the difference in the responses of the LAC9 and GAL4 proteins to GAL80. Our results indicate that this difference is due primarily to the fact that under wild-type conditions, the level of LAC9 protein in S. cerevisiae is much higher than that of GAL4, which suggests that LAC9 escapes GAL80-mediated repression by titration of GAL80 protein in vivo. The difference in response to GAL80 is not due to amino acid sequence differences between the LAC9 and GAL4 carboxyl termini. We discuss the implications of these results for the mechanism of galactose metabolism regulation in S. cerevisiae and K. lactis. PMID- 2550791 TI - Composite transposable elements in the Xenopus laevis genome. AB - Members of two related families of transposable elements, Tx1 and Tx2, were isolated from the genome of Xenopus laevis and characterized. In both families, two versions of the elements were found. The smaller version in each family (Tx1d and Tx2d) consisted largely of two types of 400-base-pair tandem internal repeats. These elements had discrete ends and short inverted terminal repeats characteristic of mobile DNAs that are presumed to move via DNA intermediates, e.g., Drosophila P and maize Ac elements. The longer versions (Tx1c and Tx2c) differed from Tx1d and Tx2d by the presence of a 6.9-kilobase-pair internal segment that included two long open reading frames (ORFs). ORF1 had one cysteine plus-histidine-rich sequence of the type found in retroviral gag proteins. ORF2 showed more substantial homology to retroviral pol genes and particularly to the analogs of pol found in a subclass of mobile DNAs that are supposed retrotransposons, such as mammalian long interspersed repetitive sequences, Drosophila I factors, silkworm R1 elements, and trypanosome Ingi elements. Thus, the Tx1 elements present a paradox by exhibiting features of two classes of mobile DNAs that are thought to have very different modes of transposition. Two possible resolutions are considered: (i) the composite versions are actually made up of two independent elements, one of the retrotransposon class, which has a high degree of specificity for insertion into a target within the other, P-like element; and (ii) the composite elements are intact, autonomous mobile DNAs, in which the pol-like gene product collaborates with the terminal inverted repeats to cause transposition of the entire unit. PMID- 2550793 TI - Beta interferon subtype 1 induction by tumor necrosis factor. AB - Tumor necrosis factor (TNF) induces an antiviral state in various cell lines. This antiviral state is quite similar to that established by interferon (IFN), e.g., TNF treatment of HEp-2 cells induces 2',5'-oligoadenylate synthetase activity. Both antiviral activity and synthetase induction are greatly reduced when TNF treatment occurs in the presence of a beta interferon subtype 1 (IFN beta 1)-neutralizing antiserum. However, no one has yet directly demonstrated IFN beta 1 induction, either as an antiviral activity in supernatants from TNF treated cells or as IFN-specific mRNA by Northern (RNA) blot analysis. We have adopted a recently described in vitro DNA amplification protocol for the detection of specific RNAs. By applying this method to RNA from HEp-2 cells, we could demonstrate increased levels of IFN-beta 1-specific transcripts after TNF treatment. Dose response and kinetics of IFN-beta 1 induction coincided with the TNF-induced antiviral state. Nuclear run-on analysis showed enhanced transcriptional activity of the IFN-beta 1 gene in TNF-treated cells. Our data substantiate a role of IFN-beta 1 as mediator of the biological activity of TNF in HEp-2 cells. PMID- 2550792 TI - Identification of four nuclear transport signal-binding proteins that interact with diverse transport signals. AB - The transport of proteins into the nucleus requires not only the presence of a nuclear transport signal on the targeted protein but also the signal recognition proteins and the nuclear pore translocation apparatus. Complicating the search for the signal recognition proteins is the fact that the nuclear transport signals identified share little obvious homology. In this study, synthetic peptides homologous to the nuclear transport signals from the simian virus 40 large T antigen, Xenopus oocyte nucleoplasmin, adenovirus E1A, and Saccharomyces cerevisiae MAT alpha 2 proteins were coupled to a UV-photoactivable cross-linker and iodinated for use in an in vitro cross-linking reaction with cellular lysates. Four proteins, p140, p100, p70, and p55, which specifically interacted with the nuclear transport signal peptides were identified. Unique patterns of reactivity were observed with closely related pairs of nuclear transport signal peptides. Competition experiments with labeled and unlabeled peptides demonstrated that heterologous signals were able to bind the same protein and suggested that diverse signals use a common transport pathway. The subcellular distribution of the four nuclear transport signal-binding proteins suggested that nuclear transport involves both cytoplasmic and nuclear receptors. The four proteins were not bound by wheat germ agglutinin and were not associated tightly with the nuclear pore complex. PMID- 2550794 TI - Comparison of filler DNA at immune, nonimmune, and oncogenic rearrangements suggests multiple mechanisms of formation. AB - Extra nucleotides (termed filler DNA) are commonly found at the junctions of genetic rearrangements in mammalian cells. The filler DNA at immune system rearrangements, which are called N regions, are generated at VDJ joints primarily by terminal deoxynucleotidyl transferase. However, the origin of filler DNA at genetic rearrangements in nonlymphoid cells is uncertain. In an analysis of more than 200 junctions that arose by circularization of transfected linear DNA (D. B. Roth and J. H. Wilson, Mol. Cell. Biol. 6:4295-4304, 1986), we found 18 junctions with extra nucleotides exactly at the point of circularization. Analysis of these 18 junctions indicated that nonlymphoid cells could add extra nucleotides to the ends of duplex DNA. The characteristics of the extra nucleotides at these junctions and at 31 other rearrangement junctions from nonlymphoid cells were quite similar, suggesting that many genetic rearrangements may pass through a stage with free DNA ends. A comparison of the filler DNA at these 49 nonimmune system rearrangements with 97 N regions derived from immune system rearrangements suggested that lymphoid and nonlymphoid cells use different mechanisms for insertion of filler DNA, as expected from the absence of detectable terminal deoxynucleotidyl transferase in cells from nonlymphoid tissues. The filler DNAs at a smaller group of 22 translocations associated with cancer had features in common with both immune and nonimmune system rearrangements and therefore may represent a mixture of these two processes. Mechanisms that might account for the presence of filler DNA in nonlymphoid cells are discussed. PMID- 2550795 TI - Dictyostelium discoideum myosin: isolation and characterization of cDNAs encoding the regulatory light chain. AB - Phosphorylation of the regulatory light chains (RMLC) of nonmuscle myosin can increase the actin-activated ATPase activity and filament formation. Little is known about these regulatory mechanisms and how the RMLC are involved in ATP hydrolysis. To better characterize the nonmuscle RMLC, we isolated cDNAs encoding the Dictyostelium RMLC. Using an antibody specific for the RMLC, we screened a lambda gt11 expression library and obtained a 200-base-pair clone that encoded a portion of the RMLC. The remainder of the sequence was obtained from two clones identified by DNA hybridization, using the 200-base-pair cDNA. The composite RMLC cDNA was 645 nucleotides long. It contained 60 base pairs of 5' untranslated, 483 bases of coding, and 102 base pairs of 3' untranslated sequence. The amino acid sequence predicted an 18,300-dalton protein that shares 42% amino acid identity with Dictyostelium calmodulin and 30% identity with the chicken skeletal myosin RMLC. This sequence contained three regions that were similar to the E-F hand calcium-binding domains found in calmodulin, troponin C, and other myosin light chains. A sequence similar to the phosphorylation sequence found in chicken gizzard and skeletal myosin light chains was found at the amino terminus. Genomic Southern blot analysis suggested that the Dictyostelium genome contains a single gene encoding the RMLC. Analysis of RMLC expression patterns during Dictyostelium development indicated that accumulation of this mRNA increases just before aggregation and again during culmination. This pattern is similar to that obtained for the Dictyostelium essential myosin light chain and suggests that expression of the two light chains is coordinated during development. PMID- 2550796 TI - Independent glucocorticoid induction and repression of two contiguous responsive genes. AB - Specific DNA sequence elements which contain binding sites for the glucocorticoid receptor mediate the action of glucocorticoid hormones on gene transcription. In glucocorticoid-inducible genes, these glucocorticoid-responsive elements behave as hormone-inducible enhancers of transcription. We have taken advantage of the bovine papillomavirus (BPV) system to test the stringency of glucocorticoid regulation of transcription. BPV episomes were constructed to contain two hormone regulated transcription units in close proximity; one transcription unit is under control of a glucocorticoid-inducible promoter (mouse mammary tumor virus) while the other is under control of a glucocorticoid-inhibited promoter (pro opiomelanocortin). Glucocorticoids independently regulated transcription of the two physically linked transcription units, irrespective of their relative orientation and of their proximity on the BPV episomes. This result contrasts with the so-called position-independent activity of enhancers and suggests that the multicomponent organization of eucaryotic promoters restricts the action of hormone-responsive regulatory elements to a specific transcription unit, thus accounting for the stringency of hormonal regulation observed in vivo. PMID- 2550797 TI - Origin auxiliary sequences can facilitate initiation of simian virus 40 DNA replication in vitro as they do in vivo. AB - Initiation of simian virus 40 (SV40) DNA replication is facilitated by two auxiliary sequences that flank the minimally required origin (ori) core sequence. In monkey cells, the replication rate of each of the four ori configurations changed with time after transfection in a characteristic pattern. This pattern was reproduced in an extract from SV40-infected monkey cells by varying the ratio of DNA substrate to cell extract; DNA replication in vitro depended on ori auxiliary sequences to the same extent as they did in vivo. Facilitation by ori auxiliary sequences was lost at high ratios of DNA to cell extract, revealing that the activity of these sequences required either multiple initiation factors or a molar excess of one initiation factor bound to ori. This parameter, together with ionic strength and the method used to measure DNA replication, determined the level of facilitation by ori auxiliary sequences in vitro. The activity of ori auxiliary sequences was not diminished in vivo or in vitro by increasing amounts of large tumor antigen. Therefore, ori auxiliary sequences promoted initiation of replication at some step after tumor antigen binding to ori. Furthermore, although cellular factors could modulate the activity of ori auxiliary sequences in vitro, these factors did not appear to involve nucleosome assembly because no correlation was observed between the number of nucleosomes assembled per DNA molecule and facilitation by ori auxiliary sequences. These results demonstrate that SV40 ori auxiliary sequences can function in vitro as they do in vivo and begin to elucidate their role in initiating DNA replication. PMID- 2550798 TI - A region internal to the coding sequences is essential for transcription of the yeast Ty-D15 element. AB - The major transcript of the yeast transposable element Ty1 has its 5' end in one delta and the 3' end in the opposite delta, the direct repeats of about 335 base pairs (bp) at each end of the element. The transcriptional initiation signals of the Ty-D15 element that give rise to this transcript were found to have a number of unusual characteristics. The 5' delta by itself, which contained the initiation site for Ty transcription, gave no detectable transcription. A region internal to the transcript in a translated part of the element and about 140 bp downstream of the 5' delta was essential for initiation of the major Ty transcript. This internal activating region (IAR) had several interesting properties. When the portion of the delta upstream of the initiation site was replaced with DNA fragments that did not by themselves act as promoters, initiation directed by the IAR still occurred at about the same position, 200 to 400 bp upstream of the IAR. If fragments containing the IAR were inverted, transcription could still occur. When 468 or 636 bp was inserted between the delta and the IAR, initiations occurred near the normal delta initiation site and in the inserted DNA. Therefore, the location and properties of transcription signals for Ty-D15 differ considerably from those expected for a yeast gene transcribed by RNA polymerase II. PMID- 2550799 TI - Regulation of postreceptor signaling in the pheromone response pathway of Saccharomyces cerevisiae. AB - alpha-Factor pheromone inhibits division of yeast a cells. After prolonged exposure to alpha-factor, the cells adapt to the stimulus and resume cell division. The sst2 mutation is known to inhibit adaptation. This report examines adaptation in scg1 (also designated gpa1) and STE4Hpl (Hpl indicates haploid lethal) mutants that exhibit constitutive activation of the pheromone response pathway. Recovery of the STE4Hpl mutant was blocked by the sst2-1 mutation, whereas recovery of the scg1-7 mutant was not completely blocked by sst2-1. These results indicate that both SST2-dependent and -independent mechanisms regulate postreceptor events in the pheromone response pathway. Down regulation of receptors in response to alpha-factor was independent of the signal that was generated in the scg1 mutant. PMID- 2550800 TI - Identification of a region within the Na,K-ATPase alpha subunit that contributes to differential ouabain sensitivity. AB - To analyze determinants within the Na,K-ATPase alpha subunit that contribute to differential ouabain sensitivity, we constructed and expressed a panel of chimeric cDNA molecules between ouabain-resistant and ouabain-sensitive alpha subunit cDNAs. When introduced into ouabain-sensitive monkey CV-1 cells, ouabain resistant rat alpha 1 subunit cDNA and chimeras in which the 5' end of ouabain sensitive human alpha 1 or rat alpha 2 subunit cDNA was replaced by the 5' end of rat alpha 1 subunit cDNA conferred resistance to 100 microM ouabain. Monkey cells transfected with the reciprocal chimeras were unable to survive selection in 1 microM ouabain. Rat alpha 2 subunit cDNA and a chimera in which the 5' end of rat alpha 1 subunit cDNA was replaced by the 5' end of rat alpha 2 subunit cDNA conferred resistance to 0.5 microM ouabain. These results suggest that determinants of ouabain resistance reside within the amino-terminal portions of the rat alpha 1 and alpha 2 subunits. Expression of chimeric alpha subunit cDNAs should prove useful for elucidating the structural basis of Na,K-ATPase function. PMID- 2550801 TI - Hematopoietic lineage-specific heterogeneity in the 5'-terminal region of the chicken proto-myb transcript. AB - Comparison of the nucleotide sequence of the upstream c-myb exon UE3 with the sequences of a thymus c-myb cDNA and of a B-lymphoma c-myb cDNA suggested the existence of T- and B-cell-specific heterogeneity in the 5'-terminal region of the c-myb coding sequence. This possibility was investigated with T-cell-specific and B-cell-specific DNA probes in a Northern (RNA) blot analysis of mRNAs from different hematopoietic cell types and from chicken embryo fibroblasts. The hematopoietic tissues analyzed were bone marrow, bursa of Fabricius, and thymus from 1-day-old chicks, 13-day yolk sac, and spleen from 16-day embryos. At least three different c-myb mRNA species were found to have 5'-terminal heterogeneity that was specific for either B cells, T cells, or the other hematopoietic cells and chicken embryo fibroblasts. This lineage-specific heterogeneity in the c-myb transcript was found to be expressed in the bone marrow precursors of B and T cells before they migrated to their definitive differentiation sites. S1 nuclease protection analysis of the UE3 exon, part of which appeared to be coding sequences for thymic c-myb mRNA, revealed that this exon is utilized either in its entirety or partially in a cell-lineage-specific manner by all six tissues analyzed. Also, the 5'-terminal exon(s) present in the thymus cDNA was absent in c-myb mRNAs from the other cell types analyzed. PMID- 2550802 TI - Micronuclear genome organization in Euplotes crassus: a transposonlike element is removed during macronuclear development. AB - After mating, hypotrichous ciliated protozoa transform a set of their micronuclear chromosomes into thousands of short, linear DNA molecules that form the macronuclear genome. To examine micronuclear genome organization in the hypotrich Euplotes crassus, we have analyzed two cloned segments of micronuclear DNA as well as the macronuclear DNA molecules that are derived from them. E. crassus was found to display a number of features characteristic of other hypotrich genomes, including (i) clustering and close spacing of the precursors of macronuclear DNA molecules, (ii) the frequent occurrence of internal eliminated sequences within macronuclear precursors, (iii) overlapping macronuclear precursors, (iv) lack of telomeric repeats at the ends of macronuclear precursors, and (v) alternative processing of the micronuclear chromosome to yield multiple macronuclear DNA molecules. In addition, a moderately repetitive, transposonlike element that interrupts the precursors of two macronuclear DNA molecules has been identified and characterized. This transposonlike element, designated Tec1, is shown to be reproducibly removed from one of the macronuclear precursors during independent episodes of macronuclear development. PMID- 2550803 TI - Identification of HeLa cell nuclear factors that bind to and activate the early promoter of human polyomavirus BK in vitro. AB - Human polyomavirus BK (BKV), an oncogenic DNA virus, differs from other papovaviruses in the organization of the regulatory region and in tissue tropism for kidney cells. The noncoding regulatory region of the viral genome in prototype strains includes three 68-base-pair (bp) repeats, each containing a number of potential regulatory elements. Some of these signals are unique to human papovaviruses, and others are homologous to those identified in many viral and cellular genes. We evaluated the contribution of individual 68-bp repeats to the initiation of transcription from the early promoter in a HeLa cell extract and identified cis-acting elements to which human cellular factors bind to activate transcription. The early promoter with only one copy of the 68-bp repeat could accurately initiate transcription in vitro, but additional copies were required for its stimulation. DNA-binding assays and DNase I protection experiments identified six domains in the regulatory region protected by human cellular factors. Two of these footprints were located within the proximal and distal 68-bp repeats, and one was located at the late side of the repeats. These footprints were centered over a TGGA(N)5-6GCCA core and were produced by a protein of the nuclear factor 1 (NF-1) family. This protein is either identical or similar to that which binds to the high-affinity site at the origin of adenovirus DNA replication. Three other domains, two at the junctions of the 68 bp repeats and one in the late side of the repeats, were partially protected by proteins with AP-1- and Sp-1-like activities. Transcription initiation from the early promoter was drastically reduced when a complete 68-bp repeat or the NF-1 binding site was used as a competitor in the in vitro assay. However, a point mutation within the NF-1 binding site, which reduced NF-1 binding in vitro to a level comparable to that of nonspecific DNA, also eliminated its ability to compete with early transcription. The murine homolog of the AP-1 binding site had a modest effect on in vitro transcription. Our results suggest that, among the multiple HeLa cell nuclear factors, NF-1 acts as a major activator of the early promoter in vitro. PMID- 2550804 TI - Simian virus 40 DNA replication in vitro: identification of multiple stages of initiation. AB - A cell-free DNA replication system dependent upon five purified cellular proteins, one crude cellular fraction, and the simian virus 40 (SV40)-encoded large tumor antigen (T antigen) initiated and completed replication of plasmids containing the SV40 origin sequence. DNA synthesis initiated at or near the origin sequence after a time lag of approximately 10 min and then proceeded bidirectionally from the origin to yield covalently closed, monomer daughter molecules. The time lag could be completely eliminated by a preincubation of SV40 ori DNA in the presence of T antigen, a eucaryotic single-stranded DNA-binding protein (replication factor A [RF-A]), and topoisomerases I and II. In contrast, if T antigen and the template DNA were incubated alone, the time lag was only partially decreased. Kinetic analyses of origin recognition by T antigen, origin unwinding, and DNA synthesis suggest that the time lag in replication was due to the formation of a complex between T antigen and DNA called the T complex, followed by formation of a second complex called the unwound complex. Formation of the unwound complex required RF-A. When origin unwinding was coupled to DNA replication by the addition of a partially purified cellular fraction (IIA), DNA synthesis initiated at the ori sequence, but the template DNA was not completely replicated. Complete DNA replication in this system required the proliferating cell nuclear antigen and another cellular replication factor, RF-C, during the elongation stage. In a less fractionated system, another cellular fraction, SSI, was previously shown to be necessary for reconstitution of DNA replication. The SSI fraction was required in the less purified system to antagonize the inhibitory action of another cellular protein(s). This inhibitor specifically blocked the earliest stage of DNA replication, but not the later stages. The implications of these results for the mechanisms of initiation and elongation of DNA replication are discussed. PMID- 2550806 TI - Fusion of adenovirus E1A to the glucocorticoid receptor by high-resolution deletion cloning creates a hormonally inducible viral transactivator. AB - The 289-amino-acid E1A protein of adenovirus type 2 stimulates transcription from early viral and certain cellular promoters. Its mechanism is not known, and there exist no temperature-sensitive mutants of E1A that could help to elucidate the details of E1A transcriptional activation. To create for E1A such a conditional phenotype, we fused portions of E1A to the human glucocorticoid receptor (GR) to make transactivation by E1A dependent on the presence of dexamethasone. Nested subsets of the E1A coding region, centered around the 46-amino-acid transactivating domain, were substituted for the DNA-binding domain of the GR. One of the resulting chimeric proteins (GR/E1A-99), which included the entire E1A transactivating domain, stimulated expression from a viral early promoter (E3) exclusively in the presence of hormone. GR/E1A-99 did not transactivate a GR responsive promoter. It therefore exhibited the promoter specificity of E1A while possessing the hormone inducibility of the GR. Two smaller chimeras that contained only portions of the E1A transactivating domain failed to transactivate E3. These three chimeras were constructed by a novel strategy, high-resolution deletion cloning. In this procedure, series of unidirectional deletions were made with exonuclease III on each side of the E1A coding region at a resolution of 1 to 2 nucleotides. The large number of in-frame fragments present in the collection of deleted clones facilitated the construction of the GR/E1A chimeras and can be used to create many additional fusions. PMID- 2550805 TI - Mammalian growth-associated H1 histone kinase: a homolog of cdc2+/CDC28 protein kinases controlling mitotic entry in yeast and frog cells. AB - Mammalian growth-associated H1 histone kinase, an enzyme whose activity is sharply elevated at mitosis, is similar to cdc2+ protein kinase from Schizosaccharomyces pombe and CDC28 protein kinase from Saccharomyces cerevisiae with respect to immunoreactivity, molecular size, and specificity for phosphorylation sites in H1 histone. Phosphorylation of specific growth associated sites in H1 histone is catalyzed by yeast cdc2+/CDC28 kinase, as shown by the in vitro thermal lability of this activity in extracts prepared from temperature-sensitive mutants. In addition, highly purified Xenopus maturation promoting factor catalyzes phosphorylation of the same sites in H1 as do the mammalian and yeast kinases. The data indicate that growth-associated H1 kinase is encoded by a mammalian homolog of cdc2+/CDC28 protein kinase, which controls entry into mitosis in yeast and frog cells. Since H1 histone is known to be an in vivo substrate of the mammalian kinase, this suggests that phosphorylation of H1 histone or an H1 histone counterpart is an important component of the mechanism for entry of cells into mitosis. PMID- 2550807 TI - Antibodies to synthetic peptide from the residue 33 to 42 domain of c-Ha-ras p21 block reconstitution of the protein with different effectors. AB - Residues 32 to 40, which are conserved among ras proteins from different species, are likely to participate in interactions with the p21 effector system. With the goal of understanding the structural basis of the regulatory functions of c-Ha ras p21, we produced rabbit antisera against a synthetic peptide corresponding to amino acids 33 to 42 of the protein. The affinity-purified antibodies interacted specifically with p21 and with the antigenic peptide. The epitope recognized by the antibodies appeared to be centered on threonine 35. The antibodies inhibited both in vitro p21-induced production of cyclic AMP in detergent extracts of RAS defective yeast membranes and GAP-stimulated GTPase activity. However, monoclonal anti-ras antibodies Y13-259 and Y13-238 were not capable of specifically inhibiting interactions of p21 with these two putative effector proteins. The apparent inhibitory effect of Y13-259 on stimulation of p21 by GAP was due to a greatly reduced rate of exchange of nucleotides in the binding pocket of the protein. These findings provide additional support for the essential role of the residue 32 to 40 domain as the true effector site and further evidence of the involvement of GAP as a cellular effector of ras proteins. PMID- 2550808 TI - Early pre-B-cell transformation induced by the v-fms oncogene in long-term mouse bone marrow cultures. AB - Murine long-term bone marrow cultures that support B-lymphoid-cell development were infected with a helper-free retrovirus containing the v-fms oncogene. Infection of B-lymphoid cultures resulted in the rapid clonal outgrowth of early pre-B cells, which grew to high cell densities on stromal cell feeder layers, expressed v-fms-coded glycoproteins, and underwent immunoglobulin heavy-chain gene rearrangements. Late-passage cultures gave rise to factor-independent variants that proliferated in the absence of feeder layers, developed resistance to hydrocortisone, and became tumorigenic in syngeneic mice. The v-fms oncogene therefore recapitulates known effects of the v-abl and bcr-abl oncogenes on B lineage cells. The ability of v-fms to induce transformation of early pre-B cells in vitro underscores the capacity of oncogenic mutants of the colony-stimulating factor-1 receptor to function outside the mononuclear phagocyte lineage. PMID- 2550809 TI - Mutations in the hormone regulatory element of mouse mammary tumor virus differentially affect the response to progestins, androgens, and glucocorticoids. AB - Transcription of the mouse mammary tumor virus DNA is known to be induced by several steroid hormones. Using chimeric MMTV plasmids containing mutations within the hormone regulatory element, we have previously studied the regions required for the glucocorticoid response in mouse fibroblasts. Here we report the characterization of elements essential for the stimulation by progestins and androgens as compared with glucocorticoids. The same set of mutant plasmids was transfected into the human mammary tumor cell line T47D, and the specific transcripts were analyzed by an S1 nuclease protection assay. Androgen-mediated stimulation, although weak, showed an extended sensitivity to mutations, with a slight preference for the proximal region. The results with progestin suggest that sequences within all the described sites protected by the receptor in vitro are required and that the promoter-proximal region (-128 to -78 from the RNA start site) is more important than the distal one (-190 to -160). Moreover, a binding site for nuclear factor I was not required for the progestin response, whereas it was required for glucocorticoids. Thus, the various steroid receptors play a role in the differential regulation of mouse mammary tumor virus transcription by recognizing distinct sequence differences in the hormone regulatory element and interacting with different factors bound to the promoter. PMID- 2550810 TI - Homologous plasmid recombination is elevated in immortally transformed cells. AB - The levels of intramolecular plasmid recombination, following transfection of a plasmid substrate for homologous recombination into normal and immortally transformed cells, have been examined by two independent assays. In the first assay, recovered plasmid was tested for DNA rearrangements which regenerate a functional neomycin resistance gene from two overlapping fragments. Following transformation of bacteria, frequencies of recombinationlike events were determined from the ratio of neomycin-resistant (recombinant) colonies to ampicillin-resistant colonies (indicating total plasmid recovery). Such events, yielding predominantly deletions between the directly repeated sequences, were substantially more frequent in five immortal cell lines than in any of three normal diploid cell strains tested. Effects of plasmid replication or interaction with T antigen and of bacterially mediated rejoining of linear molecules generated in mammalian cells were excluded by appropriate controls. The second assay used limited coamplification of a control segment of plasmid DNA, and of the predicted recombinant DNA region, primed by two sets of flanking oligonucleotides. Each amplified band was quantitated by reference to a near linear standard curve generated concurrently, and recombination frequencies were determined from the ratio of recombinant/control DNA regions. The results confirmed that recombinant DNA structures were generated within human cells at direct repeats in the transfected plasmid and were markedly more abundant in an immortal cell line than in the diploid normal cells from which that line was derived. PMID- 2550811 TI - Effect of silencer on polyomavirus DNA replication. AB - We have cloned the cellular sequence termed box DNA from the enhancer region of polyomavirus F9 mutant fPyF9. Box DNA functions as a negative transcriptional element (silencer) in undifferentiated F9 cells but not in differentiated L cells. Plasmid DNAs containing the origin and enhancer of polyomavirus were used to measure simultaneously transcriptional and replication activities in transfected cells. DNA replication activity was significantly reduced under conditions in which the silencer was able to reduce enhancer activity in F9 cells. On the other hand, when the silencer could not repress enhancer activity in MOP-8 cells, which are mouse NIH 3T3 cells producing polyomavirus T antigen constitutively, replication activity was still intact. The silencer itself had no effect on DNA replication or transcription in either type of cells. Furthermore, the insertion of a 6-base oligonucleotide within a consensus sequence of box DNA abolished the repressive effect of the silencer on DNA replication and enhancer activities. These results suggest that enhancer factors, interacting with silencer factors, may be closely associated with the mechanism of replication. PMID- 2550812 TI - Negative transcriptional regulatory element that functions in embryonal carcinoma cells. AB - We have cloned the polyomavirus mutant fPyF9, which persists in an episomal state in F9 embryonal carcinoma cells (K. Ariizumi and H. Ariga, Mol. Cell. Biol. 6:3920-3927, 1986). fPyF9 carries three copies of exogenous sequences, the prototype of which is a 21-base-pair repeat (box DNA), in the region of the enhancer B domain of wild-type polyomavirus DNA. The consensus sequence, GCATTCCATTGTT, is 13 base pairs long. The box DNA inserted into fPyF9 appeared to come from a cellular sequence and was present in many kinds of DNAs, including F9 chromosomal DNA. The biological function of box DNA was analyzed by chloramphenicol acetyltransferase expression assays, using chimeric plasmids containing box DNA conjugated with simian virus 40 promoter elements. The results showed that box DNA repressed the activities both of the simian virus 40 promoter and enhancer only in transfected undifferentiated F9 cells and not in differentiated LTK- cells. Box DNA functioned independently of orientation and position with respect to the promoter in an enhancerlike manner, although the effect of box DNA was opposite that of the enhancer. The XhoI linker insertion into the consensus sequences of box DNA abolished the repression activity, and the protein(s) recognizing the consensus sequences was identified only in F9 cells, not in L cells. These analyses suggest that box DNA may be a negative regulatory element that functions in undifferentiated cells. PMID- 2550814 TI - Proteolytic processing of pro-ACTH/endorphin begins in the Golgi complex of pituitary corticotropes and AtT-20 cells. AB - The intracellular sites where proteolytic processing of pro-ACTH/endorphin or POMC take place have not yet been reliably identified. We have used affinity purified antisera that recognize only the products of POMC processing and immunoelectron microscopy to identify the compartments of rat pituitary corticotropes and mouse AtT-20 cells in which cleavage occurs. Immunoperoxidase labeling of cryostat sections and immunogold labeling of ultrathin frozen sections were used for localization of the processing sites. By both procedures we detected processed peptides in Golgi cisternae and secretion granules. Within the Golgi, labeling was limited to the last or transmost cisterna and was most concentrated in its dilated rims which contain condensing secretory protein. No labeling of other Golgi cisternae was seen. All Golgi cisternae were labeled, however, when antisera that recognize unprocessed POMC were used for immunolabeling. We conclude that in AtT-20 and rat pituitary cells: 1) processing of POMC through at least two endo- and exoproteolytic cleavage steps and alpha amidation of joining peptide begin in the trans Golgi subcompartment; 2) no detectable processing takes place before POMC reaches the trans Golgi cisterna; and 3) this Golgi cisterna as well as secretion granules contain the active enzymes necessary for proteolytic processing and alpha-amidation. PMID- 2550813 TI - Integration of an aberrant retrotransposon in Saccharomyces cerevisiae. AB - We describe an atypical composite Ty1 element that apparently resulted from the concurrent integration of two complete elements. A portion of the central region of one of these elements was inverted between two long terminal repeats. Inversions of this type have been detected among unintegrated retroviral circles. It now appears that such intermediates can be incorporated into the genome. PMID- 2550815 TI - A quantitative comparison of dual control of a hormone response element by progestins and glucocorticoids in the same cell line. AB - Progesterone receptor-containing T47D human breast cancer cells are responsive to progestins but fail to respond to other steroid hormones, in particular dexamethasone, because they have no measurable levels of receptors for estrogens, androgens, or glucocorticoids. To quantitatively study dual responsiveness of the mouse mammary tumor virus (MMTV) promoter to progestins and glucocorticoids, we have stably transfected T47D cells with a glucocorticoid receptor (GR) expression vector. A cloned derivative (A1-2) was isolated that expresses a normal, full length GR, as assessed by steroid binding and Western immunoblot with a monoclonal anti-GR antibody. Moreover, GR is expressed at levels (80,000-100,000 molecules per cell) comparable to the high levels of endogenous progesterone receptor (200,000 molecules per cell). In A1-2 cells transiently transfected with an MMTV-chloramphenicol acetyl transferase reporter gene, induction by glucocorticoid was substantially greater (5-fold) than induction mediated by progestins. These results suggest that glucocorticoids may be the primary regulator of MMTV. PMID- 2550816 TI - Signalling through sIgA on a novel murine B lymphoma. AB - A novel murine B lymphoma expressing membrane-associated IgA was isolated and used to compare mechanisms of signal transduction by sIgM and sIgA. Like other isotypes so far studied, crosslinking of sIgA by anti-immunoglobulin antibodies stimulates hydrolysis of inositol phospholipids and causes elevation of intracellular free calcium. Furthermore, signals generated through sIgA are coupled to elevation of c-fos proto-oncogene expression. Coupling appears to be through the protein kinase C rather than through the Ca2+ component of sIg signalling as phorbol diester, but the Ca2+ ionophore cannot mediate this effect. Thus these results, coupled with those from earlier studies, show that early signal transduction through surface immunoglobulin appears to be similar regardless of the particular isotype involved in binding ligand. PMID- 2550818 TI - Maturation of transmission in reinnervated mouse soleus muscle. AB - After the tibial nerve of the mouse was cut unilaterally and immediately resutured, reinnervation of soleus muscle proceeded rapidly and muscle isometric contraction characteristics reached normal levels within 2 months. In contrast, synaptic transmission remained immature since resistance to presynaptic (magnesium) or postsynaptic (curare) blocking solutions remained reduced. Results suggest that release probability and transmitter stores were smaller than normal. To study the effect of training, animals were allowed to run in wheels. Running caused a delay in reinnervation at 18-20 days, which was, however, abolished by 4 weeks. On the other hand, exercise counteracted development of denervation atrophy. The safety margin of transmission in runners was higher than in nonrunners at 4 weeks, indicating enhanced maturation, but was lower at 2 months of reinnervation. These results suggest that recovery of muscle precedes maturation of synaptic transmission. PMID- 2550817 TI - Colchicine blocks beta adrenoceptor and class I antigen-specific interactions. AB - Previously we have demonstrated a molecular relationship between H-2 class I antigens and beta adrenoceptors from cardiac tissue. Here we show this type of interaction taking place with beta adrenoceptors from splenic cells and their purified membranes and the participation of cytoskeletal proteins in the phenomenon. Alloimmune, as well as anti-class I but not anti-class II, antibodies were able to inhibit in a competitive manner the binding of (-) [3H]dihydroalprenolol to splenic lymphocytes and their purified membranes, and to increase cyclic AMP levels in intact cells as a consequence of beta adrenoceptor activation. Furthermore, colchicine (a microtubule disrupting drug), but not cytochalasin B (a microfilament disrupting drug), was able to abrogate alloimmune antibody inhibition over the beta radioligand binding to its receptor on both intact splenocytes and their membranes. Alloantibody actions were significantly diminished by peripheral protein solubilization in purified spleen cell membranes. These data pointed indirectly to the participation of a colchicine binding protein in class I antigen hormone-receptor associations. PMID- 2550819 TI - Acute dapsone-induced peripheral neuropathy. PMID- 2550820 TI - AAEE Minimonograph #33: electrodiagnostic approach to defects of neuromuscular transmission. AB - Clinical testing for neuromuscular dysfunction is supported by an extensive amount of excellent basic information about normal and abnormal subcellular physiology and ultrastructure. This information provides an essential frame of reference for describing the rationale of single-fiber electromyography (SFEMG). SFEMG in turn helps to explain the more conventional clinical testing of neuromuscular function by repetitive nerve stimulation (RNS). Electrical findings in myasthenia gravis, Lambert-Eaton myasthenic syndrome, and botulinum intoxication are discussed from the subcellular level via the cellular level (SFEMG) to the integrated responses of whole muscle (RNS) as a rational means of understanding the technique of clinical repetitive nerve stimulation. PMID- 2550821 TI - Ephaptic transmission in hemifacial spasm: a single-fiber EMG study. AB - In two patients with hemifacial spasm (HFS), single-fiber EMG recordings in facial muscles demonstrated low jitter in the late responses produced by stimulation of peripheral branches to other facial muscles. Surgical decompression of the facial nerve in one patient was followed by clinical improvement and disappearance of the abnormal late responses. These observations are consistent with the hypothesis that there is ephaptic transmission among peripheral branches of the facial nerve at the site of compression in HFS. PMID- 2550822 TI - Vipoma, glucagonoma and the role of Sandostatin. PMID- 2550823 TI - Regulation of Ca2+-dependent K+-channel activity in tracheal myocytes by phosphorylation. AB - Isoprenaline is a beta-adrenergic agonist of clinical importance as a remedy for asthma. In airway smooth muscle its relaxant action is accompanied by hyperpolarization of the membrane and elevation of the level of intracellular cyclic AMP. Hyperpolarization and relaxation are also induced by drugs such as forskolin, theophylline and dibutyryl cAMP, indicating that cAMP-dependent phosphorylation is involved in producing the electrical response. Cyclic AMP dependent protein kinase (protein kinase A) has been reported to activate Ca2+ dependent K+ channels in cultured aortic smooth muscle cells and snail neurons. The membrane of tracheal smooth-muscle cells is characterized by a dense distribution of Ca2+-dependent K+-channels. We have now examined the effect of isoprenaline and protein kinase A on Ca2+-dependent K+-channels in isolated smooth muscle cells of rabbit trachea, using the patch-clamp technique. Our results show that the open-state probability of Ca2+-dependent K+-channel of tracheal myocytes is reversibly increased by either extracellular application of isoprenaline or intracellar application of protein kinase A. We also show that this effect is significantly enhanced and prolonged in the presence of a potent protein phosphatase inhibitor, okadaic acid. PMID- 2550824 TI - Scintillation proximity assay. PMID- 2550825 TI - Inositol phosphates and cell signalling. AB - Inositol 1,4,5-trisphosphate is a second messenger which regulates intracellular calcium both by mobilizing calcium from internal stores and, perhaps indirectly, by stimulating calcium entry. In these actions it may function with its phosphorylated metabolite, inositol 1,3,4,5-tetrakisphosphate. The subtlety of calcium regulation by inositol phosphates is emphasized by recent studies that have revealed oscillations in calcium concentration which are perhaps part of a frequency-encoded second-messenger system. PMID- 2550827 TI - Malignant insulinoma: is combined treatment with verapamil and the long-acting somatostatin analogue octreotide (SMS 201-995) more effective than single therapy with either drug? AB - The efficacy of treatment with either verapamil (V), the long-acting somatostatin analogue octreotide (OCT) alone, and the combined treatment (V + OCT) were studied in a patient with a symptomatic metastasized malignant insulinoma. Treatment with V alone resulted in a slight increase in blood glucose levels. Treatment with OCT alone resulted in a clear increase in blood glucose levels. Treatment with V + OCT was slightly more effective than single therapy with either drug but could not prevent hypoglycaemic episodes completely. It is also shown that the effect of treatment with V and OCT alone on glucoregulatory hormones is completely different. While treatment with V did not inhibit insulin secretion, treatment with OCT clearly did. Treatment with OCT might also modify counterregulatory hormone secretion during hypoglycaemia. PMID- 2550826 TI - Glucose-inhibition of glucagon secretion involves activation of GABAA-receptor chloride channels. AB - The endocrine part of the pancreas plays a central role in blood-glucose regulation. It is well established that an elevation of glucose concentration reduces secretion of the hyperglycaemia-associated hormone glucagon from pancreatic alpha 2 cells. The mechanisms involved, however, remain unknown. Electrophysiological studies have demonstrated that alpha 2 cells generate Ca2+ dependent action potentials. The frequency of these action potentials, which increases under conditions that stimulate glucagon release, is not affected by glucose or insulin. The inhibitory neurotransmitter gamma-aminobutyric acid (GABA) is present in the endocrine part of the pancreas at concentrations comparable to those encountered in the central nervous system, and co-localizes with insulin in pancreatic beta cells. We now describe a mechanism whereby GABA, co-secreted with insulin from beta cells, may mediate part of the inhibitory action of glucose on glucagon secretion by activating GABAA-receptor Cl- channels in alpha 2 cells. These observations provide a model for feedback regulation of glucagon release, which may be of significance for the understanding of the hypersecretion of glucagon frequently associated with diabetes. PMID- 2550828 TI - Chemical integrity of [3H]GABA used in binding studies. AB - A method which is claimed to be able to determine the proportion of "true" GABA within radiolabeled GABA used in binding studies was tested using [3H]GABA. The method was found to be unsuitable for 3H-labeled GABA and, furthermore, both theoretical considerations and the present experimental data indicated that it could also produce misleading results with [14C]GABA. PMID- 2550830 TI - [A study on the protective mechanism of phenytoin in transient global ischemia]. AB - Ischemia gives rise to severe energy depletion and imbalance of Ca2+ homeostasis. This condition leads to activation of phospholipases A2, A1 and to attenuation of ATP dependent reacylation. As a result, free fatty acid (FFA) especially arachidonic acid accumulates. Phenytoin has been reported to blockade the various Ca2+ channels. In this study, we could investigate the effect of phenytoin on the liberation of FFA, energy metabolites, various nucleotides metabolism, Na+, K+ ATPase activity, and water and electrolyte contents in the ischemic brain. Inhibitory effects of phenytoin against FFA accumulation in the ischemia, and increase of parietal cortex water content in the recirculation were brought about. In addition, Na+,K+-ATPase activity in the ischemia was accelerated by phenytoin. Phenytoin may reduce intracellular Ca2+ by blocking the Ca2+ channel into the cytoplasma, or by activation of Na+-Ca2+ exchange transport system following the acceleration of Na+,K+-ATPase activity. Another conceivable way for this acceleration of Na+,K+-ATPase may be derived from the preservation of the energy state, protein metabolism, and lipid metabolism. PMID- 2550829 TI - Iron-induced lipid peroxidation and inhibition of dopamine synthesis in striatum synaptosomes. AB - Crude striatum synaptosomes (P2 fraction) from Fischer 344 female rats were incubated in the presence of ADP-chelated Fe3 (0.5-50 microM) and ascorbate (250 microM). Intrasynaptosomal conversion of tyrosine to dopamine (DA) was measured by 14CO2 evolution from L-[1-14C]tyrosine in the absence of added cofactors and DOPA decarboxylase. Malondialdehyde (MDA) was measured as an index of lipid peroxidation. A concentration-dependent inhibition of DA synthesis by ADP Fe3./ascorbate was found with 50% inhibition occurring at 2.5 microM Fe3 concentration. This was accompanied by marked accumulation of MDA. Ascorbate or ADP alone did not affect DA synthesis and ADP-Fe3 in the absence of exogenous ascorbate was effective only above 25 microM. Exogenously added MDA did not inhibit DA synthesis. Purified synaptosomes were isolated from peroxidized and control P2 actions using sucrose gradients. Membrane microviscosity of the purified synaptosomes was assessed by nitroxyl spin labels of stearic acid using electron paramagnetic resonance techniques. There was a significant increase in membrane microviscosity as a result of ADP-Fe3./ascorbate induced peroxidation. Maleimide nitroxide spin-label binding to protein sulfhydryls was significantly modified by peroxidation of striatum synaptosomes. The weakly immobilized component of the sulfhydryl spin-label (w) was drastically decreased whereas the strongly immobilized component (s) was modified less, thus leading to a marked reduction of w/s ratio. The exposure of striatum synaptosomes to the peroxidizing system resulted in a significant increase in total iron and in a 25% decrease in protein sulfhydryl content. It is concluded that iron-induced damage to the DA synthetic system is mediated by alterations of the structural properties of nerve ending membranes. PMID- 2550831 TI - [Pineal metastatic tumor from lung cancer initially caused by neurological abnormalities of pineal body tumor]. AB - A rare case is reported of pineal metastasis from lung cancer initially caused by neurological abnormalities of pineal tumor. A 70-year-old female suffering from headache and deterioration of consciousness for 1 week was admitted. She also had a tumor on both sides of her neck. On admission, neurological examination revealed disturbance of upward gaze, and CT scans showed hydrocephalus and pineal tumor. The tumor was seen as a slightly high density mass on non-contrast CT, and was homogeneously enhanced after administration of contrast material. Right V-P shunt and excision of the left neck tumor were performed at the same time. Pathological diagnosis of neck tumor was undifferentiated carcinoma metastasized to cervical lymph nodes. Extensive study was made, by bronchial fiberscope and biopsy, in order to find the origin of the malignancy and disclosed a small cell lung cancer of left lower lobe. The patient took radiation therapy for both the whole brain (60 Gy) and for the bilateral cervical regions (45 Gy). Two courses of chemotherapy using CDDP, ADR, VCR and CY were administered. Both the neck and the pineal tumors were markedly reduced in size at the termination of radiation therapy. However, she was readmitted 3 months later because of dyspnea. Chest X-P revealed enlargement of the left-lung tumor. She died on April 22, 1987. General autopsy disclosed invasive enlargement of left lung cancer, however, no remote metastasis was found. Examination of pineal region showed only necrotic pineal tissue, and no tumor cell was seen in either macroscopic or microscopic study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550832 TI - Intraneuronal eosinophilic bodies of beef cattle (Japanese brown). AB - Eosinophilic cytoplasmic inclusion bodies were observed in the midbrain, pons, medulla and spinal cord of beef cattle (Japanese Brown). The inclusion bodies, observed in 14 of 20 cases exhibiting neurological signs and in one of nine healthy cases, differed in morphology and/or staining properties from previously reported inclusion bodies in humans and animals. PMID- 2550833 TI - On the role of brain mineralocorticoid (type I) and glucocorticoid (type II) receptors in neuroendocrine regulation. AB - Administrations of the glucocorticoid receptor antagonist (anti-glucocorticoid, RU38486) and the mineralocorticoid antagonist (anti-mineralocorticoid, RU28318) followed by frequent, sequential blood sampling were employed to investigate the possible role the brain mineralocorticoid receptor (MR, type I) and glucocorticoid receptor (GR, type II) have in the regulation of basal and stress induced adrenocortical secretion in the rat. The anti-mineralocorticoid and anti glucocorticoid were administered subcutaneously (s.c.) at doses of 2.5 mg and 1.0 mg/100 g body weight, respectively. Both antagonists were also given intracerebroventricularly (i.c.v.) at a dose of 100 ng/rat. Under basal non stressed conditions (at the diurnal trough in the morning), injections of either saline, anti-glucocorticoid (s.c. or i.c.v.) or anti-mineralocorticoid (s.c.) did not have effect on the plasma corticosterone level. The anti-mineralocorticoid given intracerebroventricularly, however, caused an elevation of plasma corticosterone up to 60 min after the injection. Exposure of the rats to a novel environment resulted in a large increase in the plasma corticosterone level, which was slightly reduced in the rats treated with the anti-glucocorticoid. In vehicle-treated rats, the level returned to basal values at 90 min, while in the anti-glucocorticoid- and anti-mineralocorticoid-treated groups, it remained elevated for prolonged periods. The present study thus shows that (1) the anti glucocorticoid RU38486 via the brain GR has no effect on the basal plasma corticosterone level in the morning but interferes with a glucocorticoid negative feedback following stress and (2) the anti-mineralocorticoid RU28318 via the brain MR elevates the basal plasma corticosterone level and enhances adrenocortical secretion following stress.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550834 TI - Splanchnic nerve stimulation modulates steroid secretion in hypophysectomized dogs. AB - To test whether or not splanchnic neural input to the adrenal gland affects secretion of steroids from the adrenal cortex, the thoracic splanchnic nerve was electrically stimulated in pentobarbital-anesthetized dogs after hypophysectomy and replacement with physiological concentrations of ACTH. An adrenal vein cannula was placed to permit measurement of cortisol, corticosterone, 11 deoxycortisol, epinephrine and norepinephrine secretion rates and adrenal blood flow. Plasma ACTH was measured and the presentation rate of ACTH was calculated as the product of plasma ACTH concentration and adrenal plasma flow. Dogs were infused initially with ACTH for 60 min at 2 ng/min followed by infusion for 60 min at 10 ng/min. Within each infusion period, the distal end of the nerve was stimulated (20 V; 0.5-ms pulse duration) at 4 and at 20 Hz for 10 min each. Nerve stimulation resulted in a frequency-dependent increase in mean arterial pressure, in epinephrine and norepinephrine secretion and in adrenal blood flow. Arterial ACTH remained constant during nerve stimulation; however, increased adrenal blood flow resulted in increased presentation rate of ACTH to the adrenal. Cortisol secretion increased in response to nerve stimulation at 4 and 20 Hz during infusion of 2 and 10 ng/min ACTH and occurred prior to changes in presentation rate of ACTH. Corticosterone secretion also increased after stimulation at both frequencies, but the response was observed only during infusion of 10 ng/min ACTH. In contrast, 11-deoxycortisol decreased after nerve stimulation at 4 Hz but showed no response after stimulation at 20 Hz during infusion of 2 and 10 ng/min ACTH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550836 TI - Arginine vasopressin is a much more potent stimulus to ACTH release from ovine anterior pituitary cells than ovine corticotropin-releasing factor. 1. In vitro studies. AB - Cultured rat and ovine anterior pituitary cells were treated with a range of doses (0.01-1,000 nM) of arginine vasopressin (AVP) and ovine corticotropin releasing factor (CRF), alone or in combination, and medium and cell content of immunoreactive (ir-)ACTH determined. In rat cells, a dose-response curve to CRF was obtained, with a threshold dose of 0.1 nM; AVP was much less effective alone, but augmented CRF responses when administered with CRF. In ovine pituitary cells AVP markedly stimulated ACTH release in a dose-dependent fashion, and with a threshold of 0.1 nM; in contrast, CRF increased ACTH release over basal only at doses greater than 100 nM. In combination, subthreshold doses of AVP potentiated rat pituitary cell responses to CRF; addition of 1 nM of AVP to varying doses of CRF was more effective in terms of ACTH release than addition of 1 nM of CRF to increasing doses of AVP. In contrast, in ovine cells the addition of 1 nM CRF to increasing doses of AVP elicited a larger ACTH response than the addition of 1 nM AVP to increasing doses of CRF. Dexamethasone pretreatment (5 nM) for 48 h significantly decreased CRF potentiation of AVP-stimulated ACTH release in ovine cells. These studies confirm that CRF is a more potent stimulus of ACTH release than AVP in the rat, and establish that in contrast AVP is a much more potent stimulus of ACTH secretion than CRF in isolated ovine pituitary cells. PMID- 2550835 TI - Further studies on the stimulatory action of nicotine on adrenocortical function in the rat. AB - The aim of the present study was to further characterize the site of action of nicotine-induced hypothalamo-hypophyseal-adrenal (HHA) activation. A systemic injection of nicotine in concentrations of 65-2,100 micrograms/kg elevated serum corticosterone (CS) concentrations in a time and dose-dependent manner. Serum ACTH levels were also significantly increased. Pretreatment with dexamethasone (40 micrograms/kg) or the nicotinic antagonist, mecamylamine (1 mg/kg), abolished the ACTH and CS secretory responses to nicotine. Intracerebroventricular administration of antinicotinic acetylcholine receptor antibodies, prepared from the serum of myasthenia gravis patients, completely inhibited the nicotine induced HHA activation. Bilateral lesions of the paraventricular nucleus similarly inhibited the nicotine-induced adrenocortical activity. These results suggest that nicotine activates the HHA axis by a central mechanism which ultimately requires the integrity of the paraventricular nucleus. Moreover, these findings indicate that the nicotinic adrenocortical effect is mediated specifically through activation of central nicotinic cholinergic receptors. PMID- 2550837 TI - Adaptation of the adrenocorticotropic response to chronic intermittent stress was altered by intracerebroventricular infusion of hemicholinium-3. AB - The role of diencephalic cholinergic neurotransmission in regulating hypothalamic pituitary-adrenocortical (HPA) axis was investigated by means of administration of hemicholinium-3 (HC-3), a blocker of acetylcholine synthesis, in the third ventricle of hemispherectomized pigeons. Except for an early increase in ACTH and corticosterone levels following injection as a bolus of HC-3 that was ascribed to some stress-like situation, all data indicated that hypothalamic acetylcholine depletion resulted in inhibiting effects on the HPA axis. Twenty-four hours after injection of 6 micrograms of HC-3, the response to acute stress was markedly reduced in both magnitude and duration. Permanent instillation of HC-3 in the third ventricle at the rate of 0.25 microgram/h for 9 days led to lowered basal resting HPA activity and severely affected the development of adaptation to chronic intermittent stress. The anticipatory conditioned, endocrine response did not appear whereas attenuation of the poststress component was amplified. It is suggested that cholinergic mechanisms are involved in modulating the HPA function and particularly the conditioning process that takes place in the course of adaptation of the HPA response to chronic intermittent application of the same stressor. PMID- 2550838 TI - Effect of histamine depletion on circadian variations of corticotropin and corticosterone in rats. AB - The effects of cerebral histamine depletion induced by alpha fluoromethylhistidine (FMH) on corticotropin (ACTH) and corticosterone secretions were examined. Neither acute nor chronic FMH treatment altered the corticoadrenal responses to three types of stress: transposition, immobilization and water immersion. And exposure to stress did not affect the hypothalamic content of histamine. However, chronic intracerebral treatment with FMH had a significant effect on the circadian rhythm of the plasma corticosterone (CS) level in rats. Namely, it caused a marked attenuation of the amplitude of the peaks of the CS level resulting in an almost arrhythmic state. The maximum differences between FMH treated and untreated groups were seen at 8.00 and 20.00 h, the times when the illumination condition changed (light onset 8.00 h). This treatment with FMH also had a similar effect on the plasma ACTH concentration; namely the plasma ACTH level in the saline treated group was lower than that of the FMH-treated group at light onset and higher than the latter at dark onset, but was similar to the latter at other sampling points. These results indicate the histaminergic modulation of the circadian rhythm of hormonal secretion of the adrenal cortex and show that this phenomenon is mediated through the central nervous system by an influence on the rhythm of hypophyseal ACTH secretion possibly through alteration in the concentration of corticotropin-releasing factor. PMID- 2550840 TI - Testosterone modulates oxytocin binding in the hypothalamus of castrated male rats. AB - Oxytocin (OT) binding sites are modulated by estrogens in several brain regions including the ventromedial hypothalamic nucleus (VMN) in both male and female rats. To further study steroid regulation of OT receptor binding, we examined the effect of androgen replacement in castrated male rats on OT binding with quantitative autoradiographic methods. Castrated adult male rats were treated with either 250 micrograms testosterone propionate (TP) or oil for 2 days and killed 48 h after the last injection. Brain sections through the preoptic area and VMN were labeled with 5.0 nM[3H]-OT +/- 5.0 microM unlabeled OT or 1.0 microM[Thr4,Gly7]OT and apposed to tritium-sensitive film for 7 weeks. Results of this study show that TP increased [3H]-OT binding up to 5-fold in the ventrolateral VMN and 4-fold in the bed nucleus of the stria terminalis. In addition [Thr4,Gly7]OT completely displaced [3H]-OT binding in the VMN indicating that binding in this brain region was specific to OT receptors. Because estrogens also increase OT receptor binding in male rats, it is possible that TP affects OT binding after being converted by aromatase to estradiol. PMID- 2550839 TI - Estradiol modulation of oxytocin binding in the ventromedial hypothalamic nucleus of male and female rats. AB - It has been suggested that estradiol and oxytocin (OT) may interact as neuroendocrine components in the regulation of sexual behavior. In the present study the effect of estradiol benzoate (EB) treatment (50 micrograms/kg body weight/2 days) on [3H]-OT binding was evaluated in adult and 21-day-old gonadectomized male and female rat brains. Coronal sections through the ventro medial nucleus of the hypothalamus (VMN) were analyzed in three different section planes. EB priming induced an increase in [3H]-OT binding in the VMN of both male and female rats. Greater binding site density and significant EB effects were found in the most caudal plane where the ventrolateral portion of the VMN is well defined at both ages. OT binding in the central amygdaloid nucleus was not affected by this treatment but higher binding levels were found in the most caudal sections irrespective of hormonal status or sex. No sex differences were detected in OT binding in the VMN of basal or EB-treated animals. These results suggest that a dose of EB which activates female sexual behavior in female but not in male rats is able to induce similar levels of OT binding in the VMN of animals of both sexes. PMID- 2550841 TI - Phosphoinositide hydrolysis induced by depolarization and sodium channel activation in mouse cerebrocortical slices. AB - Carbachol, a muscarinic receptor agonist and the sodium channel-activating agents, scorpion venom, veratridine, batrachotoxin and aconitine, were shown to stimulate the formation of [3H]inositol phosphates in [3H]inositol-labelled miniprisms, obtained from the cerebral cortex of the mouse. The inositol response to the Na+ channel-activating agents was inhibited by the sodium channel blocker tetrodotoxin (TTX), while the response induced by carbachol was partially resistant to TTX. The response to scorpion venom and the TTX-insensitive portion of the response to carbachol was additive, indicating different mechanisms. The presence of high potassium (K+) induced hydrolysis of inositide in a TTX insensitive manner and was not additive with that resulting from sodium channel activators, thus indicating a common mechanism. The addition of large concentrations of magnesium to block the release of acetylcholine, did not inhibit the inositol response to high K+ or to veratridine. Calcium channel blockers such as nickel or cobalt, or the dihydropyridine calcium (Ca2+) channel activator BAY K 8644 and the calcium channel blocker nifedipine, nimodipine or PN 200 110 had little effect. Monensin, a sodium ionophore, stimulated the turnover of phosphatidylinositol at non-depolarizing concentrations and the omission of Na+ ions inhibited the response to sodium channel agents and to high K+. Thus, membrane potential and gradients of K+, Na+ and Ca2+ are all important factors determining the final effect on the turnover of phosphatidylinositol. The data are consistent with a model in which all these factors impinge on the Na+/Ca2+ exchanger regulating internal Ca2+ that, in turn, activates phospholipase C. PMID- 2550842 TI - Haloperidol pretreatment unmasks the kappa opioid effects of U-50, 488H on cortical EEG and EEG power spectra in rats. AB - Adult female Sprague-Dawley rats were implanted with chronic cortical EEG electrodes and intravenous cannulae. U-50, 488H injection (5.0 mg/kg) produced initial EEG desynchrony and EEG spectral power that was mainly distributed over the zero to 10 Hz range, including a relatively small spectral peak in the 4-6 Hz band. In contrast, following haloperidol pretreatment (0.1 mg/kg), U-50, 488H injection produced high-voltage EEG bursts and a predominant EEG spectral peak in the 4-6 Hz band. These effects of U-50, 488H after haloperidol pretreatment were identical to those previously demonstrated with the benzomorphan kappa agonist ethylketocyclazocine. Thus, haloperidol pretreatment unmasked the kappa opioid effects of U-50, 488H. PMID- 2550843 TI - Jejunal proabsorptive actions of selective opiate agonists administered via the cerebral ventricles. AB - Non-selective enkephalin analogs, such as [D-Ala2, Met5]enkephalinamide, have been found to increase jejunal absorption in the rat and dog after their intracerebroventricular administration. In the present investigation, experiments were designed to characterize the brain opiate receptor subtype mediating this action in the rat proximal jejunum in situ and to assess the involvement of extrinsic sympathetic nerves innervating this gut region in opioid-induced absorption. Changes in jejunal water transport were examined in rats pretreated with bolus i.c.v. doses (less than or equal to 1 microgram) of the respective mu , delta-, or kappa-opiate agonists [D-Ala2, N-Me-Phe4, Gly5-ol]enkephalin (DAGO), [D-Pen2, D-Pen5]enkephalin or U-50488 and infused intravenously with the secretagogue prostaglandin E1 (PGE1, 5 micrograms/kg-min). In saline-pretreated rats, PGE1 produced large, time-dependent decreases in jejunal fluid absorption. Of the opiate agonists examined, only DAGO in submicrogram doses significantly inhibited PGE1 actions 60 to 90 min after its administration. Extirpation of the celiac and superior mesenteric ganglia, major sources of sympathetic neurons innervating the upper gut, significantly attenuated both the antiabsorptive actions of PGE1 and the proabsorptive actions of DAGO (0.3 micrograms, i.cv.). These results suggest that CNS mu-opiate receptors modulate jejunal absorption in the rat, an action mediated through extrinsic sympathetic nerves innervating the upper small intestine. PMID- 2550844 TI - Molecular characteristics and peptide specificity of bradykinin binding sites in intact neuroblastoma-glioma cells in culture (NG 108-15). AB - Here we report that the mouse neuroblastoma-glioma hybrid cell line NG108-15 possess high-affinity binding sites for the nonapeptide bradykinin, as revealed by competitive displacement of 125I-8Tyr bradykinin by various bradykinin analogs. These binding sites were further characterized by covalent cross-linking of 125I-8Tyr bradykinin to intact NG108-15 grown as a monolayer, using dithiobis succinimidylpropionate (DTSP) as a cross-linking reagent. Sodium dodecyl sulfate (SDS) electrophoresis after solubilization of the cross-linked cells, demonstrated the preferential and specific labeling of two polypeptides with apparent molecular weights of Mr = 36,000 and Mr = 47,000. A third polypeptide of Mr = 69,000 was labeled less intensely. PMID- 2550845 TI - Topical organization of the projections of neurons of the substantia nigra and the ventral field of the tegmentum of the midbrain to the head of the caudate nucleus of the cat. PMID- 2550846 TI - Restructuring of the topical organization of the motor cortex of the rat following damage to the opposite hemisphere. PMID- 2550847 TI - Modulation of the transmission of signals of the rabbit septum in the presence of influences on the cholinergic system. PMID- 2550849 TI - [The surgical therapy of stage-III pulmonary cancer. Comments on 71 cases]. PMID- 2550848 TI - Cerebral ultrasound images in prenatal cytomegalovirus infection. AB - A male newborn with prenatal cytomegalovirus infection was referred for cranial ultrasound. The cranial ultrasound demonstrated areas of increased echogenicity in the thalamic and gray nuclei resembling "a branched candlestick". Doppler technique located the "branched candlestick" along the thalamostriate arteries. This image is particularly interesting because to our knowledge it has never before been described in congenital cytomegalovirus infection, but only in congenital rubella. PMID- 2550850 TI - [Pulmonary carcinoma. The effect of assessing histologic type on the prognosis]. PMID- 2550851 TI - [Adjuvant surgery of limited-disease pulmonary microcytoma and the role of TNM classification. A review of the authors' own experience]. PMID- 2550852 TI - Precise targeting of the pathology of the sialoglycoprotein, PrP, and vacuolar degeneration in mouse scrapie. AB - Widespread immunostaining of PrP protein was demonstrated in scrapie mouse brain, distributed diffusely in the neuropil and focally in amyloid plaques, microglia and 2-5 microns structures resembling neuronal processes. With the 87V scrapie strain, which produces focal vacuolation in particular areas, PrP pathology was precisely targeted to these same areas, predating vacuolar degeneration by at least several weeks. On the other hand, both vacuolar and PrP changes were widely distributed throughout the brain with the ME7 scrapie strain. It is likely that the precise targeting of PrP pathology, followed by vacuolar degeneration, reflects an underlying targeting and localised replication of infectious agent. PMID- 2550853 TI - Effects of age on binding sites for calcitonin gene-related peptide in the rat central nervous system. AB - The binding site distribution of calcitonin gene-related peptide (CGRP) was studied in the central nervous system of aged rats (22 months old) and compared with that of young rats (2 months). The regional distribution of [125I]Tyr-rat CGRP binding in coronal sections of young and old rat CNS was examined by an in vitro autoradiographic technique. The results, showed that in aged rats there was a marked reduction in CGRP binding, without any change in binding affinity, in the hippocampus, the nucleus rhomboideus, the nucleus arcuatus, the colliculus superior, the substantia grisea centralis and the spinal cord. In the cortical areas, the amygdala, the caudatus putamen and the accumbens binding was not modified. In the cortex cerebellaris CGRP binding was strikingly greater in the aged rats. The increase in binding might be a consequence of an adaptive process due to a decline of the peptide synthesis with age and is suggestive of a role for CGRP in the cerebellum functions. PMID- 2550854 TI - Phosphoinositides and GTP binding proteins involved in muscarinic generation of hippocampal rhythmic slow activity. AB - We examined the role of phosphoinositide turnover in muscarinic rhythmic slow activity (RSA; also called theta) in rat CA3 pyramidal neurons. Pre-incubation of hippocampal slices in pertussis toxin (which inhibits some GTP-binding proteins) or in Li+ (which blocks inositol phosphate degradation, and thereby decreases the resynthesis of phosphoinositides), prevented the induction of RSA by carbachol. Phorbol esters, which can activate protein kinase C (PKC) directly, did not induce RSA but inhibited muscarinic RSA. We infer that muscarinic RSA involves a GTP-binding protein linked increase in phosphoinositide turnover, while the activation of PKC may have a negative feedback role. PMID- 2550855 TI - Peculiarities of receptor-channel complexes for inhibitory mediators in the membranes of lamprey spinal cord neurones. AB - Effects of inhibitory mediators on the membranes of isolated lamprey spinal cord neurones were investigated by means of whole-cell recording and concentration clamp techniques. Glycine and gamma-aminobutyric acid (GABA) applications evoked desensitizing chloride currents. The concentrations at half-maximum effects were 16 microM for glycine- and 1.5 mM for GABA-activated currents. Ionic responses to applications of both amino acids demonstrated full cross-desensitization. Strychnine and bicuculline suppressed both glycine- and GABA-activated conductances to the same degree. We suggest that in the membranes of lamprey spinal cord neurones glycine and GABA act on the same receptor-channel complexes. PMID- 2550856 TI - Comparison of low-threshold Ca2+ currents in the hippocampal CA1 neurons among the newborn, adult and aged rats. AB - The electrical and pharmacological properties of the low-threshold Ca2+ current were compared among the newborn, adult and aged rats using the isolated hippocampal CA1 pyramidal neurons. The current-voltage relationship and the time constant of the decay phase of the low-threshold Ca2+ current in adult and aged rats were similar to those in newborn rats. The low-threshold Ca2+ current of adult and aged rats was also sensitive to nicardipine, a dihydropyridine derivative, same as that of newborn rats. We concluded that the nature of low threshold Ca2+ current in the rat hippocampal CA1 pyramidal neurons is not affected by the aging. PMID- 2550857 TI - Endocrine nursing. Diet in the treatment of diabetes mellitus. PMID- 2550859 TI - Testicular malignancy associated with massive inguinal adenopathy and life threatening hemorrhage. PMID- 2550858 TI - Pulmonary oncocytoma. PMID- 2550860 TI - Resistance to antiviral drugs at last. PMID- 2550861 TI - Iron, copper and zinc levels in serum and cirrhotic liver of patients with and without hepatocellular carcinoma. AB - It has been pointed out that hepatocellular carcinoma (HCC) develops more frequently in cirrhotic liver with siderosis than in liver without iron deposition, that excess copper in hepatocytes inhibits hepatocarcinogenesis, and that an increase in copper and a decrease in zinc are seen in the sera of patients with various malignant tumors. Iron, copper and zinc concentrations in the serum and liver were estimated in normal subjects and cirrhotic patients with and without HCC. Serum copper level was significantly higher in cirrhotic patients with or without HCC than in normal subjects. No significant differences were observed in the levels of these trace elements in the serum and liver of cirrhotic patients with and without HCC. The current study seems to indicate that iron, copper and zinc do not play an important role in the development of HCC in cirrhotic patients in Japan. PMID- 2550862 TI - Cytogenetic analysis of Wilms' tumor: evidence of a multistep process. AB - We report the cytogenetic findings observed in the cancer cells of a Wilms' tumor in a 11-month-old girl without the phenotype of the WAGR. Two populations were identified: one had a 46,XX,del(11)(p11p13) karyotype and the other had a translocation involving the deleted chromosome 11 and chromosome 12, t(11;12)(p11;q13). This result strongly suggests that the deletion of 11p was the initiating event to the tumor and that the translocation has been instrumental in promoting its development. PMID- 2550863 TI - In situ detection of Epstein-Barr virus markers in nasopharyngeal carcinoma patients. AB - Cryosections of nasopharyngeal tissue from 9 patients suspected of having nasopharyngeal carcinoma (NPC) were examined for the presence of Epstein-Barr virus (EBV) markers in situ to assess virus infection in the nasopharynx. Viral DNA, EB nuclear antigen, and/or early antigens (EA.D and EA.R) were detected in 5 NPC specimens. EBV infection was not confined to the tumor areas of the biopsy specimens. Lymphoid cells and nontumor areas of these specimens contained EBV markers. In addition, nasopharyngeal tissues obtained from 3 of 4 patients in clinical remission for the disease showed evidence of EBV infection. PMID- 2550864 TI - Modulation of adenylate cyclase activity in bovine lens epithelial cells. AB - Adenylate cyclase activity of bovine lens epithelial cells is activated by guanine nucleotides or fluoride. Additional activation to that of guanylylimidodiphosphate [Gpp(NH)p] was induced by isoproterenol. Insulin was found to inhibit adenylate cyclase activity. Adenosine at low concentrations (5 50 nM) increased adenylate cyclase activity, while at high concentrations it lowered the activity of the enzyme. The occurrence of R and P adenosine sites in lens epithelial cells is suggested. Like other tissues, lens epithelial cells possess a transduction system (adenylate cyclase-G protein complex) for external messages. PMID- 2550865 TI - Regulation of protein adenosine diphosphate ribosylation in bovine lens during aging. AB - Adenosine diphosphate-ribosyltransferase (ADPRT) and poly-ADP-ribose glycohydrolase (poly-ADPRG) activities were investigated in the different structures of bovine lenses. These activities and protein ADP ribosylation were detected only in the lens epithelium proliferative layer. The poly-ADPR glycohydrolase activity decreased, but the poly-ADPR polymerase activity increased during aging. In lens epithelial cells of older animals, the poly-ADP ribose level and average chain length decreased but the chain number increased. Involvement of DNA break accumulation during aging in the increase of ADPRT activity of lens epithelial cells is discussed. PMID- 2550866 TI - Sphingomyelinase activity in the bovine and human eyes. AB - We examined biochemically sphingomyelinase activity in the bovine and human ocular tissues, using trinitrophenylaminododecanoylsphingosylphosphorylcholine as substrate. The enzyme activity in the crude extracts of neuroretina, retinal pigment epithelial cells, and optic nerve of the bovine eyes was proportional to protein concentration. The activities in these tissues were little activated with Mg2+ at pH 5.1 but was activated with Mg2+ at pH 7.5. The enzyme activity at pH 5.1 was also detected in the iris and ciliary body, neuroretina, retinal pigment epithelium and choroid, and optic nerve of the human eyes. It was highest in the macula, compared with other areas of the human ocular fundus. PMID- 2550867 TI - Bacterial corneal ulcers among Arabs in Kuwait. AB - A retrospective clinicomicrobiological study of 228 patients with bacterial corneal ulcers was performed. Positive cultures of corneal ulcer samples were obtained from 68% of all patients. A high incidence of Staphylococcus epidermidis was isolated from patients' ulcers, Staphylococcus aureus, Pseudomonas, Streptococcus pneumoniae, and Streptococcus were the next most frequent pathogens. Of some help in identifying the causative organism were the locations of the ulcer, the presence or absence of hypopyon and the frequency of perforation of the ulcer. Cephaloridine, gentamicin and polymyxin were found to be the most effective therapy. Gram strains of ulcer samples were positive for organisms in only 13% of patients and accordingly were not considered useful in determining initial therapy of ulcers. Since corneal and conjunctival cultures in the majority of the control group were negative, one can presume that most of the S. epidermidis isolated from patients' ulcers was exogenous in nature. PMID- 2550868 TI - The risk of enucleation after proton beam irradiation of uveal melanoma. AB - Enucleation after proton beam irradiation of uveal melanomas occurred in 64 (6.4%) of 994 eyes with a median follow-up time of 2.7 years. The median time between irradiation and enucleation in the 64 enucleated eyes was 13 months. The probability of retaining the eye was 95 and 90%, 2 and 5 years postirradiation, respectively. Three percent of eyes were enucleated during posttreatment year 1, and the yearly rate was 1% by the fourth year. No patient had enucleation later than 5 1/2 years posttreatment. The complication most likely to result in enucleation was neovascular glaucoma although this was frequently managed without enucleation. Other common reasons for enucleation were documented or suspected tumor growth and complete retinal detachment with associated loss of vision. The leading risk factors for enucleation were anterior tumor margin involving the ciliary body, tumor height greater than 8 mm, and proximity of the tumor to the fovea. Based on the presence or absence of these factors, 5-year eye retention rates were 99, 92, and 76% for low-, moderate-, and high-risk groups, respectively. Thus, the probability of eye retention after proton beam irradiation is high even among those at greatest risk of enucleation. PMID- 2550870 TI - Effect of smoked tobacco tar on the growth, cytolytic action, DNA synthesis, and gene expression of herpes simplex virus. AB - Previous experiments have shown that topical application of smoked tobacco tar condensate to HSV-infected oral mucosae of mice promotes neoplastic changes. Although HSV is an oncogenic virus, the virus must be inactivated and lose its cytolytic activity to be oncogenic in vitro and in vivo. Therefore, we investigated the effect of the water-extractable components of smoked tobacco tar condensate (smoked tobacco tar extract) on the growth, cytolytic activity, DNA synthesis, and gene expression of type 1 herpes simplex virus (HSV-1). Diluted tar extracts in tissue culture medium markedly inhibited the replication and cytolytic activity of HSV-1 in vitro. The smoked tobacco tar extract also notably inhibited the synthesis of viral DNA and late gene expression in a dose-dependent manner. The synthesis of alpha- and beta-classes of viral proteins was not affected by the extract. Therefore, it is probable that the smoked tobacco tar extract inhibits the synthesis of viral DNA directly rather than indirectly by depressing early gene expression. PMID- 2550869 TI - Lack of correlation between HLA-B35 resistance against herpes labialis and antibody titers to HSV-1. AB - To investigate whether genetic factors linked to the human leukocyte antigens (HLA) might influence individual resistance to recurrent herpes labialis (RHL), we studied the frequencies of HLA-A, -B, and -C antigens in a sample of Sicilian population. The frequency of HLA-B35 was significantly decreased in the patient group (p corrected = 0.018). Consequently, the relative risk of development of RHL in a subject positive for HLA-B35 was 20 times smaller than in a subject who does not bear B35. Furthermore, a study was made of the possible relationship between the presence of HLA-B35 antigen and antibody titers to herpes simplex virus type 1 (HSV-1) in 62 persons affected or not by RHL. Significantly elevated titers to HSV-1 were found in the diseased group (p less than 0.001), but the geometric mean of antibody titers in HLA-B35 positive subjects was not different from that of B35 negative subjects either of RHL-affected or in healthy persons. These data are in a good agreement with the hypothesis that humoral immune responses play a marginal role in the protection from HSV-1 recurrences. PMID- 2550871 TI - Incomplete polymerization of Cavalite with the use of recommended photopolymerization times: a warning of possible cytotoxic effects. AB - As part of a study of the suitability of new materials for use as a retrofilling material, we examined the polymerization properties of Cavalite, a light-cured, hydroxyapatite and glass ionomer-containing cavity liner. By varying the time of photopolymerization, it was found that polymerization for 20 to 30 seconds according to the manufacturer's recommendations is not sufficient to ensure complete polymerization. The implications of this incomplete polymerization are discussed in terms of possible cytotoxic effects on tissues exposed to unpolymerized Cavalite, both when used in retrofilling situations and as a deep cavity liner. PMID- 2550872 TI - E6 protein of human papillomavirus type 18 binds zinc. AB - The E6 open reading frames of human and animal papillomaviruses encode a transforming protein containing a conserved pattern of repeating cysteine doublets (cys-x-x-cys) similar to that found in steroid receptor zinc finger proteins. The spacing between the cysteine doublets, however, is twice as long as in any other zinc finger protein. To demonstrate that an E6 protein could indeed bind zinc, we synthesized the human papillomavirus type 18 E6 protein in insect cells with a baculovirus vector and analysed the protein for zinc-binding activity by a zinc-blot assay. Probing of E6 protein blotted to nitrocellulose from SDS-polyacrylamide gels with radioactive [65Zn]Cl2 demonstrated that it possessed zinc-binding activity. Reduction of cysteines with DTT prior to the addition of zinc greatly increased the zinc-binding activity of blotted E6 protein. Competition experiments showed that Cd2+ and Co2+ were more effective competitors for zinc-binding than Mg2+ or Ca2+, indicating that zinc atoms may be tetrahedrally coordinated in E6-zinc complexes. We mapped zinc-binding protein domains by proteolytic cleavage and demonstrated that a small 4kDa fragment of the protein retained zinc-binding activity, consistent with a model of individual 29 amino acid zinc-binding 'fingers'. PMID- 2550873 TI - V-sis gene (SSV) is expressed transiently and independently of v-gag (SSAV) after infection of fibroblasts with SSV/SSAV. AB - To investigate the mechanism of cell transformation by the retroviral v-sis gene, we examined the mode of its mRNA expression after infection of primate fibroblasts with Simian Sarcoma Virus (SSV/SSAV). Surprisingly transient expression of the 5.3 kb transcript of v-sis was detected between day two and four after infection. Addition of cycloheximide did not reverse the down regulation of v-sis expression. Suramin, which uncouples the PDGF receptor complex, had no effect on the pattern of v-sis expression. A marginal but non transient expression of c-myc and c-fos mRNA upon v-sis expression was detected. Studies on nuclear run-off and m-RNA stability suggest that the half-life time of v-sis mRNA is about 8 h or longer and that its expression is controlled rather by transcriptional than by post-transcriptional mechanisms. The up and down regulation of v-sis expression is independent of the expression of the helper virus (SSAV) gag-genes. This indicates that v-sis oncogene (SSV) and structural genes of the helper virus (SSAV) are obviously under separate expression control. PMID- 2550874 TI - Destruction of normal human eosinophils by Entamoeba histolytica. AB - We evaluated the in-vitro interaction of normal human eosinophil leucocytes and a virulent strain of Entamoeba histolytica (HM1-IMSS) in the presence of immune serum. At a 10:1 (eosinophil:amoeba) ratio a significant time-dependent destruction of eosinophils was found from the first hour onward, and a similar, albeit weaker, cytopathic effect was found in the 200:1 ratio mixtures, with some delay in the microscopic evidence of such effect. Results were unaffected by serum factors, and amoebae emerged virtually unharmed throughout these experiments, again regardless of the presence of serum factors. These results indicate that, as with neutrophil leucocytes, virulent E. histolytica is capable of destroying normal human eosinophils in vitro. PMID- 2550875 TI - [In vitro activity of 5 new quinolones against Gardnerella vaginalis]. AB - The minimal inhibitory concentrations (MICs) of five new quinolones were determined by agar dilution, for 50 clinical isolates of Gardnerella vaginalis. They are compared with those of metronidazole, ampicillin, erythromycin and tetracycline which are widely used for the treatment of lower genital tract infections in women. The MICs of rosoxacin and pefloxacin are high, but those of ofloxacin (MIC 50, 2 mg/l), fleroxacin (MIC 50, 2 mg/l) and ciprofloxacin (MIC 50, 1 mg/l) are lower. They can explain the effectiveness observed with this latter antibiotic in vivo, and allow their clinical experiment. PMID- 2550876 TI - Elastonectin and the elastin receptor. AB - Two different types of interactions were described between cells and elastin: 1) the adhesion of cells to insoluble fibrous elastin and 2) the binding of soluble elastin derived peptides by cells. We could show that these are two different mechanisms underlying those two types of interactions. The adhesion of cells to elastin fibers is mediated by a cell-membrane complex with a 120 kD protein as the main adhesive compound which we proposed to designate elastonectin. Three other proteins (67, 60 and 45 kD) were coisolated with elastonectin. Adhesion of insoluble elastin to mesenchymal cells (fibroblasts, smooth muscle cells) is inducible with soluble elastin peptides. Highly metastatic Lewis lung carcinoma cells and human melanoma cells also exhibited the adhesion mechanism, but without a lag phase (constitutive adhesion). Binding curve (Scatchard plots) obtained with radiolabelled elastin peptides indicated the presence of high affinity elastin receptors on mesenchymal cells and human white blood cells (monocytes and PMNs) with Kd-s in the nanomolar range. The binding of elastin peptides triggers several cellular reactions such as chemotaxis, Ca++ influx (increase of Ca++i) and with mononuclear blood cells release of lytic enzymes and oxygen radicals. All these cells which exhibited elastin receptor function also exhibited adhesion although the two processes could be inhibited selectively. The receptor action is mediated by a G-protein-phospholipase-IP3 mechanism, involved in the increase of intracellular Ca++. It appears that this action also triggers the biosynthesis and membrane localization of elastonectin. The coupling of these 2 mechanisms between receptor and adhesion appears to be modified in transformed cells. PMID- 2550877 TI - Distribution of antibodies to a causative agent of exanthem subitum (human herpesvirus-6) in healthy individuals. AB - The transfer of IgG antibodies to a causative agent of exanthem subitum (human herpesvirus-6) from mother to infant was examined with an indirect immunofluorescence assay. Of 20 mothers, 85% had the antibody more than 1:10, and the significantly higher level of the antibody was found in the cord blood with a positive rate of 95%. A mean ratio of cord blood to maternal antibody titer was 1.63. A total of 301 sera from healthy individuals was examined for the age specific prevalence of antibody to the virus. In the first 2 months of life, 87% of infants had the antibody; the positive rate and the level of antibody decreased during the first 6 months of life with the lowest positive rate of 6% at 4 to 5 months of age. After 6 months of age, they increased gradually and reached the highest level at 1 year of age with the positive rate of 86%. From 2 years of age, the prevalence of the antibody was almost stable (69% to 76%) and similar to those in adolescents and adults. PMID- 2550878 TI - Cytomegalovirus retinitis in an infant with acquired immunodeficiency syndrome. AB - A case of cytomegalovirus retinitis in an infant with acquired immunodeficiency syndrome (AIDS) is described. Although well recognized as an ocular manifestation of AIDS in adults, only one case of the necrotic retinitis caused by cytomegalovirus has been described in a child with AIDS. Intravenous treatment with ganciclovir resulted in substantial ocular improvement, despite the advanced nature of the disease in one eye in which there was also secondary neovascular glaucoma. Home maintenance treatment was used via Broviac catheter. The patient later died following pulmonary infection with Pneumocystis carinii. PMID- 2550879 TI - [Clinico-laboratory analysis of the diarrhea syndrome in young children excreting rotaviruses with feces]. AB - A total of 404 children admitted to the infectious hospital of the non-intestinal type were examined with the aid of counter immunoelectroosmophoresis. Rotavirus antigen was detected in feces in 46.8% of 126 children admitted with a diagnosis of diarrhea, in 10.8% of 278 without diarrhea, and also in 32.9% of 149 children with diarrhea that occurred at the hospital. The clinical course of acute rotavirus gastroenteritis appeared to be materially influenced by acute respiratory viral infection, particularly influenza, and by certain types of opportunistic bacteria contained in high concentrations by the intestine. PMID- 2550880 TI - Voltage and temperature dependence of normal and chemically modified inactivation of sodium channels. Quantitative description by a cyclic three-state model. AB - (1) Voltage clamp experiments were done on single myelinated nerve fibres of the frog, Rana esculenta, with 10 mM TEA in the external solution to block potassium channels. (2) The potential dependence of normal sodium current inactivation was studied over a potential range of V = -50 mV to 80 mV. At depolarizations (V greater than or equal to 30 mV) inactivation is diphasic. The relative contribution of the fast phase increases from 0.4 at V = 30 mV to 0.96 at V = 80 mV. At resting potential (V = 0 mV) recovery from inactivation shows a sigmoidal time course. At strong hyperpolarization (V = -50 mV) recovery is diphasic with a predominant fast phase. (3) For a quantitative description of these findings a cyclic three-state model of inactivation, with one open and two closed states, is formulated. The potential dependence of the rate constants is determined and calculations from this model are compared with the experimental data. (4) To test the availability of the proposed model, normal inactivation was modified by treatment with 0.6 mM chloramine-T. This substance causes inactivation to become slow and incomplete; the potential dependence of steady-state inactivation becomes non-monotonic. All these effects are explained as quantitative changes of the rate constants in the cyclic inactivation model. (5) The influence of temperature on normal inactivation was studied at a range of 8-20 C. Both time constants as well as the two inactivation components are temperature-dependent. For a quantitative description of temperature effects by the cyclic model, the activation enthalpies of the rate constants are evaluated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550881 TI - Diminished toxicity of ouabain in the hypertrophied rat heart. AB - The responsiveness to ouabain of hypertrophied rat hearts has been investigated either in vivo using an isolated Langendorff rat heart perfused at various external calcium concentrations, or in vitro on purified sarcolemma vesicles. (i) The physiological study shows that at 0.25 mM CaCl2, the positive inotropic effect of 10(-5) M ouabain was diminished in hypertrophied hearts (p less than 0.02). At 0.5 mM CaCl2, the drug has no effect in controls, but it has a slight positive inotropic effect in hypertrophied hearts. At 2.50 mM CaCl2, ouabain has a negative inotropic effect accompanied by extrasystoles in controls, but in hypertrophied hearts it still has a positive inotropic effect and is not arrhythmogenic. (ii) After the pretreatment of the hearts with 2.5 mM CaCl2, the responsiveness of the (Na+, K+)-ATPase activity to ouabain was studied: the sarcolemma from hypertrophied heart contains half as many low affinity forms of (Na+, K+)-ATPase for ouabain (35% +/- 6) than in controls (80% +/- 2). Assuming that the low affinity forms are responsible for the toxic effect, these data correlate well with some of the physiological findings and suggest that the diminished toxicity for ouabain in hypertrophied hearts rather reflects a modification of the properties of the (Na+, K+)-ATPases than a change in the myocardial calcium metabolism. PMID- 2550883 TI - Effects of calmidazolium and dibutyryl cyclic AMP on the longitudinal internal resistance in sinus node strips. AB - The effects of calmidazolium (10(-7) mol/l) and db cAMP (10(-3) mol/l) on the longitudinal internal resistance (ri) in the rabbit sinus node strips were studied by means of microelectrode and extracellular recording techniques. Calmidazolium (CDZ) decreased ri by 27%. An addition of db cAMP to the CDZ containing Tyrode potentiated this effect by further 14%. db cAMP initially decreased ri by 17%, then it enhanced this resistance by 7% above the pretreatment control value. An addition of CDZ to the db cAMP-containing Tyrode reduced ri by 41% below the pretreatment control. Possible mechanisms underlying these effects were discussed in terms of cAMP, Cai, and calmodulin action on the cell coupling. PMID- 2550882 TI - 3H-ouabain binding sites in porcine skeletal muscle as influenced by environmental temperature and energy intake. AB - The influence of environmental temperature and energy intake on 3H-ouabain binding sites in skeletal muscle has been investigated in young growing pigs at 8 weeks of age. Animals lived for several weeks at 35 or 10 degrees C on a high (H) or low (L) level of energy intake. The four treatment groups were thus: 35H, 35L, 10H and 10L. The total number of 3H-ouabain binding sites (Bmax) in longissimus dorsi muscle (mean values +/- SEM) were 221 +/- 66, 214 +/- 61, 350 +/- 76 and 486 +/- 114 pmol/g wet weight for the 35H, 35L, 10H and 10L groups respectively. Bmax was significantly greater in those living in the cold than the warm (P less than 0.001). Moreover, at 10 degrees C energy intake had a significant effect, with Bmax being greater in the 10L than the 10H group (P less than 0.005). Level of energy intake had no influence on Bmax at 35 degrees C. The apparent dissociation constant was not affected by either temperature or intake. The elevated Bmax and hence the increase in number of Na+,K+-pumping sites in the cold is probably related to increased muscular activity associated with shivering. However, thyroid status also influences the number of Na+,K+-pumping sites and this may have been a contributory factor in the present study. In addition, the elevated Bmax suggests a greater potential for non-shivering thermogenesis associated with increased Na+,K+-ATPase concentration in the cold. Differences in relative stage of development between the four groups may help to explain the results for Bmax in relation to level of energy intake. PMID- 2550884 TI - Tetrodotoxin differentially blocks peak and steady-state sodium channel currents in early embryonic chick ventricular myocytes. AB - Single ventricular myocytes were dissociated from 3-day-old embryonic chick hearts and maintained in culture for 9-21 h. The whole-cell patch clamp method was used to record tetrodotoxin (TTX)-sensitive fast Na+ currents. The peak Na+ current recorded at -20 mV ranged from 10 to 70 microA/cm2. At more negative potentials, a component of the current decayed very slowly, resulting in a significant steady-state or "late" Na+ current. The origin of the late Na+ current was revealed through the examination of single Na+ channel currents recorded in outside-out membrane patches. The single Na+ channel conductance was 20 pS. A high percentage of the trials (approximately 16%) displayed multiple reopenings of a single Na+ channel, resulting in bursts of current lasting for greater than or equal to 150 ms. The frequency distributions of the Na+ channel open-times were bi-exponential. The burst-like mode of Na+ channel activity (which underlies the slowly- or non-inactivating currents recorded macroscopically), was blocked to a greater degree by TTX, compared to the peak current. The results suggest that differential blockade may occur as a result of the slow binding and increased affinity of TTX to the open Na channel. PMID- 2550885 TI - Expression of voltage-dependent sodium and transient potassium currents in an identified sub-population of dorsal root ganglion cells acutely isolated from 12 day-old mouse embryos. AB - The electrophysiological properties of a subset of dorsal root ganglion (DRG) neurons microdissected from 12-day-old (E12) mouse embryos and acutely isolated were analyzed as soon as 3 h after their isolation. Two classes of neurons were defined according to their mean diameter. The larger diameter class was examined in this study. They display uniform cytoskeletal properties with co-expression of vimentin and neurofilament triplet proteins. Patch-clamp methods also revealed a homogeneous and limited repertoire of ionic channels that included (1) a TTX sensitive Na+ current whose properties are similar to that reported in mature mammalian neurons, and (2) two types of K+ currents that can be compared with the delayed rectifier (Ik) and the transient (IA) potassium currents found in other mammalian preparations. It may be possible to use this in vitro model to examine the development of new types of currents, such as Ca2+ currents during neuronal growth and differentiation. PMID- 2550886 TI - Patch-clamp study of cultured human sweat duct cells: amiloride-blockable Na+ channel. AB - The reabsorptive duct of the eccrine sweat gland has a large transepithelial conductance consisting mainly of a high conductance to Cl- and a smaller, amiloride-blockable Na+ conductance (Bijman and Fromter 1986; Quinton 1986). Cells have been cultured from sweat ducts and their properties previously studied in Ussing chambers (Pedersen 1988) and with microelectrodes (Jones et al. 1988). We have now studied the ion channels present in excised, inside-out patches of human cultured sweat duct cells, and find a marked predominance of linear, 15 pS, amiloride-blockable, low selectivity, Na+ channels. Such channels were seen in 54/92 (59%) of the patches, with up to 7 channels recorded in a single patch. Other channel types were seen at much lower densities. The prevalence of an amiloride-blockable Na+ channel in cultured duct cells clearly distinguishes these cells from cultured sweat gland secretory cells, which lack such a channel. PMID- 2550887 TI - Effect of high NaCl intake on Na+ and K+ transport in the rabbit distal convoluted tubule. AB - Morphological studies have demonstrated that a chronic increase in distal Na+ delivery causes hypertrophy of the distal convoluted tubule (DCT). To examine whether high NaCl-intake also causes functional changes in the well defined DCT, we measured transmural voltage (VT), lumen-to-bath Na+ flux (JNa(LB], and net K+ secretion (JK(net] in DCTs obtained from control rabbits and those on high NaCl intake diets. The lumen negative VT was significantly greater in the high NaCl group than in the control group. The net K+ secretion (pmol mm-1 min-1) was greater in the high NaCl-intake group (54.1 +/- 13.0 vs 14.7 +/- 5.6). The K+ permeabilities in both luminal and basolateral DCT membranes, as assessed by the K+-induced transepithelial voltage deflection inhibitable with Ba2+, were increased in the experimental group. The lumen-to-bath 22Na flux (pmol mm-1 min 1) was also greater in the experimental group (726 +/- 119 vs 396 +/- 65). The VT component inhibitable with amiloride was also elevated in the high NaCl-intake group. Furthermore, Na+-K+-ATPase activity of the DCT was higher in the experimental than in the control group. We conclude that high NaCl intake increases both Na+ reabsorption and K+ secretion by the DCT. This phenomenon is associated with an increased Na+-K+-ATPase activity along with increased Na+ and K+ permeabilities of the luminal membrane, and an increase in the K+ permeability of the basolateral membrane. Cellular mechanisms underlying these functional changes remain to be established. PMID- 2550889 TI - Recovery from charge immobilization in sodium channels of the frog node of Ranvier. AB - The gating current off-response of the frog node of Ranvier shows a fast and a slow phase, reflecting presumably charges moving back into the resting position and charges returning from immobilization. The paper describes measurements of the time constant of the slow component, tau off2, at different potentials at 20 or 17 degrees C. The time constant tau off2 decreased markedly when the potential to which the fibre was repolarized at the end of the test pulses was decreased from -100 to -145 mV. tau off2 was compared with tau re gat and tau re Na, the time constants of the recovery of gating current and peak sodium current from the effect of a depolarizing conditioning pulse. The three time constants were equal at -145 mV, but somewhat different at -100 to -130 mV, the sequence being tau re gat greater than tau off2 greater than tau re Na. The inequality tau re gat greater than tau off2 was small and statistically not significant. It does not seem to be due to contamination of the charge movement with ionic currents because a) the Qoff/Qon ratio was near 1.0 at -100 to -120 mV, b) partial replacement of internal CsCl by KCl did not significantly alter tau off2 or Qoff/Qon, c) tau off2 was independent of pulse size. The small inequality tau re gat greater than tau off2 suggests that charges which have returned from immobilization are not immediately available for displacement. The inequality tau re gat greater than tau re Na was larger and statistically significant at -100 and -110 mV.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550888 TI - The thyroid cell monolayer in culture. A tight sodium absorbing epithelium. AB - When cultured on collagen coated nitrocellulose filters, thyroid epithelial cells form morphologically and functionally polarized monolayers. The bioelectric parameters of these monolayers were measured after mounting in Ussing chambers; transepithelial potential (Vab), short circuit current (Isc) and transepithelial resistance were respectively 12 +/- 1 mV (apical side negative), 3.8 +/- 0.2 microA cm-2 and 3250 +/- 214 omega cm2 (mean +/- SEM, n = 75). Eighty two percent of the short circuit current was related to sodium absorption as shown by inhibition by apical amiloride (Km = 0.2 microM) and by basal ouabain (K1/2 = 0.3 microM). Amphotericin B (5-25 micrograms/ml) added to the apical bath increased Isc suggesting an apical rate-limiting step. Step by step replacement of choline by Na+ in a Na+-free medium resulted in a progressive increase in Vab and Isc with half maximal effect at 20 +/- 1 mM Na+. Thyrotropin (TSH) increased Isc and Vab in a biphasic way with a transient maximum after 5 min and a plateau after 20 min (about four times the basal level at 100 microU/ml TSH). This increase in sodium transport was also inhibited by apical amiloride. Thus, in culture, the thyroid cell monolayer behaves as a tight sodium absorbing epithelium controlled by TSH, with a rate limiting apical sodium channel as the entry mechanism and a basolateral Na+, K+-ATPase as the electromotive force. PMID- 2550890 TI - Modulation produced by nifedipine of the unitary Ba current of dispersed smooth muscle cells of the rabbit ileum. AB - Using a whole-cell clamp technique on longitudinal smooth muscle cells of the rabbit ileum, it was found that the rate of decay of the macroscopic Ba current evoked by depolarization (to +20 mV) was not modified by changing the holding potential from -60 mV to -30 mV. Cadmium (20 microM) left only a small transient inward current. Using the patch clamp technique with cell-attached configuration, only one type of unitary Ba current, with conductance of 25 pS was obtained in 100 mM Ba solution. The conductance depended on the external Ba concentration, had a dissociation constant of 19 mM and maximum conductance of 42 pS, suggesting that the Ca channel is of the L-type. Depolarizing pulses (to 0 mV and 150 ms duration) delivered at a frequency of 0.5 Hz (high frequency) evoked a unitary Ba current with low open probability (p less than 0.3). However 65-75% of depolarizing pulses evoked no unitary Ba current ("blank" sweep). On the other hand, depolarizing pulses at 0.033 Hz (low frequency) reduced the number of "blank" sweeps (20-50%), and increased the fraction of sweeps with high open probability (p greater than 0.5). Thus, the channel activity may depend on the stimulus frequency. Nifedipine, in both stimulus conditions, reduced the open probability of the channel due to an increase in the fraction of "blank" sweeps to a greater extent than to a decrease in the time constant of open-times. PMID- 2550891 TI - The effect of 4-aminopyridine on action potential shape and firing pattern of rat dorsal root ganglion neurones. PMID- 2550892 TI - Effects of potassium channel blockers on differentiation of Dictyostelium discoideum. PMID- 2550893 TI - A subfragment of the beta tropomyosin gene is alternatively spliced when transfected into differentiating muscle cells. AB - A subgenomic fragment of the chicken beta tropomyosin gene which contains two alternative exons flanked by common exons was isolated and placed under the control of the SV 40 early promoter. This construction was subsequently used to transfect quail myoblasts together with a Neomycin resistance gene, and to isolate stable transfectants. mRNAs were isolated before and after differentiation and analyzed using a modification of the primer extension method. We show that myoblasts accumulate transcripts which contain the non muscle specific exon joined to the common exons while myotubes accumulate transcripts containing the muscle specific exon. These results, therefore demonstrate that such a subgenomic fragment contains all the necessary information to direct a correct developmentally regulated mutually exclusive splicing. They also strongly suggest that trans acting factors must be involved in the switch of the splicing pattern which takes place during the transition from myoblasts to myotubes. The same regulation cannot be faithfully reproduced during transient expression, since no difference in the use of exons 6A/6B is observed during differentiation and two aberrant minor splicing products are obtained which contain or lack both exons. We suggest that failure of exon 6A to splice to exon 6B is due to the existence of some structural constraints which lower the efficiency with which the intron between them is excised. PMID- 2550894 TI - Identification of sequences regulating the transcription of a Dictyostelium gene selectively expressed in prespore cells. AB - There has been considerable debate about the relative contributions of transcriptional and post-transcriptional mechanisms to the regulation of prespore gene expression in Dictyostelium. We have determined the DNA sequence upstream of D19, the Dictyostelium gene encoding PsA, a prespore-specific, cell surface protein of unknown function. Our analysis of gene fusions, in which D19 upstream sequences are placed adjacent to a heterologous reporter gene, indicates that transcriptional signals alone are sufficient for the correct temporal and cell type specific expression of this gene. We also show that the 5' and 3' boundaries of the minimal sequences necessary for correct developmental regulation lie within the region 338 to 122 nucleotides upstream of the start site of transcription but that flanking sequences seem to be necessary for optimal expression. PMID- 2550895 TI - Tet repressor binding induced curvature of tet operator DNA. AB - Tet repressor dimer binds to two tet operator sites spaced by 30 bp in the Tn10 encoded tet regulatory DNA. The effect of repressor binding on the gel mobility of circular permutated DNA fragments containing either one or both operator sequences is reported. The EcoRI induced bending of DNA is used to compare the results with other protein binding induced structural perturbations of DNA. Tet repressor bends a DNA fragment with a single tet operator to an angle of 42 degrees +/- 7 degrees. The apparent bend angle of DNA fragments containing the tandem tet operator arrangement occupied by two Tet repressor dimers turns out to be 52 degrees +/- 9 degrees. These results are interpreted with respect to the end to end distances of the bent DNA fragments. They indicate that either the intervening tet regulatory DNA between the operators or the bound operator sequences themselves contain additional perturbations from the canonical B-DNA structure. This finding is discussed in the light of previously obtained results from CD, neutron scattering, and electrooptical studies. PMID- 2550896 TI - Oligonucleotide-targeted degradation of U1 and U2 snRNAs reveals differential interactions of simian virus 40 pre-mRNAs with snRNPs. AB - We have investigated the roles of U1 and U2 snRNP particles in SV40 pre-mRNA splicing by oligonucleotide-targeted degradation of U1 or U2 snRNAs in Xenopus laevis oocytes. Microinjection of oligonucleotides complementary to regions of U1 or U2 RNAs either in the presence or absence of SV40 DNA resulted in specific cleavage of the corresponding snRNA. Unexpectedly, degradation of U1 or U2 snRNA was far more extensive when the oligonucleotide was injected without, or prior to, introduction of viral DNA. In either co-injected or pre-injected oocytes, these oligonucleotides caused a dramatic reduction in the accumulation of spliced SV40 mRNA expressed from the viral late region, and a commensurate increase in unspliced late RNA. When pre-injected, two different U2 specific oligonucleotides also inhibited the formation of both large and small tumor antigen spliced early mRNAs. However, even when, by pre-injection of a U1 5' end-specific oligonucleotide, greater than 95% degradation of the U1 snRNA 5' ends occurred in oocytes, no reduction in early pre-mRNA splicing was observed. In contrast, the same U1 5' end oligonucleotide, when added to HeLa splicing extracts, substantially inhibited the splicing of SV40 early pre-mRNA, indicating that U1 mRNP is not totally dispensable for early splicing. These findings confirm and extend our earlier observations which suggested that different pre-mRNAs vary in their requirements for snRNPs. PMID- 2550898 TI - Nucleotide sequence of cDNA encoding mouse cytochrome c oxidase subunit Va. PMID- 2550897 TI - Helper activity for gene expression, a novel function of the SV40 enhancer. AB - In this investigation we demonstrate that the enhancer of SV40 possesses an additional function which is a 'helper activity' for a more efficient transfer of viral DNA from the cytoplasm into the nucleus. After DNA transfection into rat-2 cells, the rate of CAT gene expression linked to SV40 promoter/enhancer (pSV2CAT) was approximately 50 fold higher than linked to the tk promoter (pBLCAT2). After direct nuclear microinjection this difference was reduced to a factor of 10. However cytoplasmic injection of the same number of DNA molecules/cell showed again a 50 fold increase for the SV40 promoter/enhancer-CAT construct. This difference was not due to selective degradation of the pBLCAT2 DNA. The 'helper function' did not require the intact 72 bp sequence. In vitro synthesized enhancers lacking certain enhancer motifs (GT-I, TC-II and TC-I sequence) were still effective after cytoplasmic injection whereas an 8 bp deletion (representing a part of the AP-I motif) on the downstream side strongly reduced the helper function after cytoplasmic injection but not the classical transcriptional enhancement after direct nuclear transfer. PMID- 2550899 TI - Nucleotide sequence of the gene coding for the peplomer protein (= spike protein) of infectious bronchitis virus, strain D274. PMID- 2550900 TI - Nucleotide sequence of human mitochondrial cytochrome c oxidase II cDNA. PMID- 2550901 TI - An improved method for photographing restriction endonuclease gel electrophoresis patterns of bacterial genomes. PMID- 2550902 TI - Stereospecific relationships between elements in an SV40/adenovirus-2 heterologous promoter. AB - Spacing mutations have been constructed between an SV40 72 bp enhancer element and the Adenovirus-2 major late promoter upstream element (Ad2MLP-UE), the Ad2MLP UE and the Ad2MLP TATA-box and the SV40 72 bp enhancer and the TATA-box in the absence of the Ad2MLP-UE. A clear periodic pattern of transcription from the Ad2MLP was obtained by inserting multiples of half of a DNA turn between the SV40 enhancer and the Ad2MLP-UE. We interpret this as a stereospecific requirement for the spatial alignment between the enhancer element and either the Ad2MLP-UE or the TATA-box. Insertions between the upstream element and the TATA-box did not give a periodic transcription pattern, though insertion of half of a DNA turn increased the steady-state level of specific RNA. Unexpectedly, spacing mutations between the enhancer and TATA-box in which the upstream element had been point mutated gave indications of stereospecificity, whereas the same insertions with a deleted upstream element showed little requirement for a precise stereoalignment. PMID- 2550903 TI - Characterization of 5' truncated transposed copies of the I factor in Drosophila melanogaster. AB - I factors in Drosophila melanogaster are transposable elements structurally related to Mammalian LINEs. Their transposition is activated at high frequencies during I-R hybrid dysgenesis and is associated with the production of mutations of various sorts. Very few of these mutations have been studied at the molecular level; those reported so far result either from chromosomal rearrangements or from insertions of complete I factors. We have analysed three I-R induced yellow mutations and have found that one of them is due to the insertion of an I element very similar to the complete I factor, whereas the other two are due to insertions of I elements that are truncated at their 5' ends; one of them exhibits an unusual 3' end. We discuss possible mechanisms of production of such modified I elements. PMID- 2550904 TI - DNA supercoiling determines the activation energy barrier for site specific recombination by Tn21 resolvase. AB - A kinetic analysis of site specific recombination by Tn21 resolvase has been carried out using DNA substrates of varying superhelicities. The rates for the formation of the recombinant product increased with increasing superhelicity up to a maximum value, after which further increases in superhelicity caused no further increase in rate. The reactions with DNA of reduced superhelicity were extremely slow, yet they eventually led to virtually all of the substrate being converted to product. Hence, the level of DNA superhelicity must determine the activation energy barrier for at least one of the steps within the reaction pathway that can be rate-limiting. In the presence (but not in the absence) of Mg2+ ions, the DNA was fully saturated with resolvase whenever the protein was in stoichiometric excess over resolvase binding sites on the DNA. Thus the process affected by DNA supercoiling cannot be coupled to the binding of resolvase. Instead, the step whose rate is determined by supercoiling seems to be located within the reaction pathway after the synapse. However, these reactions may involve two forms of the synaptic complex that are converted to the recombinant product at different rates. PMID- 2550905 TI - Nucleotide sequence of a plasmid born streptothricin-acetyl-transferase gene (sat 1). PMID- 2550906 TI - Sequence of a cDNA specifying subunit VIIa of human cytochrome c oxidase. PMID- 2550907 TI - [Are there regional differences in Tc uptake?]. AB - The 99mTcO4-uptake (TcTU) was measured in 363 clinically and by laboratory findings euthyroid patients without history of hyper- or hypothyroidism, no evidence of iodine contamination and with a Delta-TSH less than 2.5 IU under a longterm suppressive thyroxine therapy. The medians of TcTU in the different regional groups from the Federal Republic of Germany (mail zip code 2000-8000) were not statistically different from each other, indicating that under suppression with thyroxine the effect of the iodine supply on TcTU is nearly negligible, and that, using the same method, results from different nuclear medicine departments should not differ significantly from each other. PMID- 2550908 TI - Cerebral arterial chemotherapy. PMID- 2550909 TI - Managing peripheral neuropathy: a patient offers suggestions. PMID- 2550910 TI - Discrete mapping of brain Mu and delta opioid receptors using selective peptides: quantitative autoradiography, species differences and comparison with kappa receptors. AB - The opioid peptides, [3H]DAGO and [3H]DPDPE, bound to rat and guinea pig brain homogenates with a high, nanomolar affinity and to a high density of mu and delta receptors, respectively. [3H]DAGO binding to mu receptors was competitively inhibited by unlabelled opioids with the following rank order of potency: DAGO greater than morphine greater than DADLE greater than naloxone greater than etorphine much greater than U50488 much greater than DPDPE. In contrast, [3H]DPDPE binding to delta receptors was inhibited by compounds with the following rank order of potency: DPDPE greater than DADLE greater than etorphine greater than dynorphin(1-8) greater than naloxone much greater than U50488 much greater than DAGO. These profiles were consistent with specific labelling of the mu and delta opioid receptors, respectively. In vitro autoradiographic techniques coupled with computer-assisted image analyses revealed a discrete but differential anatomical localization of mu and delta receptors in the rat and guinea pig brain. In general, mu and delta receptor density in the rat exceeded that in the guinea pig brain and differed markedly from that of kappa receptors in these species. However, while mu receptors were distributed throughout the brain with "hotspots" in the fore-, mid- and hindbrain of the two rodents, the delta sites were relatively diffusely distributed, and were mainly concentrated in the forebrain with particularly high levels within the olfactory bulb (OB), n. accumbens and striatum. Notable regions of high density of mu receptors in the rat and guinea pig brain were the accessory olfactory bulb, striatal "patches" and "streaks," amygdaloid nuclei, ventral hippocampal subiculum and dentate gyrus, numerous thalamic nuclei, geniculate bodies, central grey, superior and inferior colliculi, solitary and pontine nuclei and s. nigra. Tissues of high delta receptor concentration included, OB (external plexiform layer), striatum, n. accumbens, amygdala and cortex (layers I-II and V-VI). Delta receptors in the guinea pig were, in general, similarly distributed to the rat, but in contrast to the latter, the hindbrain regions such as the thalamus, geniculate bodies, central grey and superior and inferior colliculi of the guinea pig were apparently more enriched than the rat. These patterns of mu and delta site distribution differed dramatically from that of the kappa opioid sites in these species studied with the peptide [125I]dynorphin(1-8). PMID- 2550911 TI - A potent and selective agonist for NK-2 tachykinin receptor. AB - Replacement of the glycine in position 8 of the C-terminal heptapeptide NKA(4-10) with beta-alanine give rise to a potent and selective agonist for the NK-2 tachykinin receptor. The affinity of [beta-Ala8]-NKA(4-10) to the NK-2 receptor is enhanced by almost one order of magnitude as compared to NKA(4-10), while affinity decreases at about the same extent at NK-1 and NK-3 receptors, respectively. Synthesis and biological activities of a series of NKA(4-10) analogues systematically replaced in each position with beta-alanine are also reported. PMID- 2550912 TI - Receptors for sensory neuropeptides in human inflammatory diseases: implications for the effector role of sensory neurons. AB - Glutamate and several neuropeptides are synthesized and released by subpopulations of primary afferent neurons. These sensory neurons play a role in regulating the inflammatory and immune responses in peripheral tissues. Using quantitative receptor autoradiography we have explored what changes occur in the location and concentration of receptor binding sites for sensory neurotransmitters in the colon in two human inflammatory diseases, ulcerative colitis and Crohn's disease. The sensory neurotransmitter receptors examined included bombesin, calcitonin gene related peptide-alpha, cholecystokinin, galanin, glutamate, somatostatin, neurokinin A (substance K), substance P, and vasoactive intestinal polypeptide. Of the nine receptor binding sites examined only substance P binding sites associated with arterioles, venules and lymph nodules were dramatically up-regulated in the inflamed tissue. These data suggest that substance P is involved in regulating the inflammatory and immune responses in human inflammatory diseases and indicate a specificity of efferent action for each sensory neurotransmitter in peripheral tissues. PMID- 2550913 TI - Further characterization of Helix FMRFamide receptors: kinetics, tissue distribution, and interactions with the endogenous heptapeptides. AB - The biphasic binding of 125I-daYFnLRFamide to crude brain membranes of Helix aspersa is due to two discernible sites (high and low affinity) rather than different agonist-induced states. The tissues in the snail that show the greatest specific 125I-daYFnLRFamide binding are the brain, reproductive system, and digestive system. The heart shows moderate binding levels, whereas low values are obtained in the oviduct and retractor muscles. The N-terminal SAR of the Helix heptapeptides (X-DPFLRFamide) indicates that, although the substitution of Leu for Met accounts for some, the dipeptide X-Asp produces most of the loss in potency at FMRFamide receptors in Helix brain. PMID- 2550914 TI - Repeated central administration of alpha-MSH does not alter the antipyretic effect of alpha-MSH in young and aged rabbits. AB - alpha-Melanocyte-stimulating hormone is a potent antipyretic when administered centrally or peripherally; however, there are no previous data on development of tolerance to the antipyretic action of this neuropeptide. In previous research, aged rabbits were more sensitive to low doses of alpha-MSH than young rabbits. We tested the antipyretic action of alpha-MSH in young and old rabbits after twice daily injections of the peptide for 10 days. Neither aged nor younger rabbits showed altered responses to alpha-MSH. This lack of tolerance underscores the importance of alpha-MSH to physiological control and survival of the host. PMID- 2550915 TI - Identification of B2 bradykinin binding sites in the guinea pig nasal turbinate. AB - Specific high affinity BK binding sites in the nasal turbinate of the guinea pig have been demonstrated. Specific [3H]BK binding (10-330 pM) was saturable, and nonlinear least squares analysis indicated the presence of a high affinity binding site with a Kd value of 60 (50-78) pM and a Bmax value of 13.1 = 2.0 fmol/mg protein. In inhibition experiments, D-Phe7-BK (a B2 antagonist) inhibited [3H]BK binding with a Ki value of 23 nM, while des-Arg9[Leu8]-BK (a B1 antagonist) had no effect up to a concentration of 10 microM. These studies indicate the presence of B2 BK receptors in the guinea pig nasal turbinate. PMID- 2550916 TI - Fibroblast and myofibroblast participation in malignant fibrous histiocytoma (MFH) of bone. Ultrastructural study of eight cases with immunohistochemical support. AB - Eight malignant fibrous histiocytomas (MFH) of bone were studied with immunohistochemistry and electron microscopy. Ultrastructurally, fibroblasts and myofibroblasts were the main tumor cells in four cases and abundant in two other cases; these cells showed immunohistochemical positivity to alpha 1-antitrypsin, vimentin and anti-muscle antigen (HHF 35). Moreover, histiocytic-like tumor cells were electron-microscopically detected in four cases, being the main tumor cell type in two of the cases; immunohistochemically these cells expressed positivity to alpha-1-antichymotrypsin (A1ACT), alpha-1-antitrypsin (A1AT) and vimentin. Present results confirm the cellular heterogeneity of MFH of bone, in which fibroblasts and myofibroblasts transformed cells play a histogenetical role, suggesting the existence of close links with classic fibrosarcoma of bone. PMID- 2550917 TI - Is the profile of binding of a panel of neoglycoproteins useful as a diagnostic marker in human lung cancer? AB - Histomorphological evaluation of the profile of carbohydrate-binding proteins with specificities to alpha- and beta-glucosides (maltose, cellobiose), alpha-L fucosides (L-fucose), alpha-D-mannosides (D-mannose), a N-acetylated sugar, present in natural glycoconjugates (N-acetyl-D-glucosamine), two negatively charged sugars (glucuronic acid and sialic acid), rhamnose (6-deoxy-L-mannose) or to heparin has been assessed. 133 cases of formalin fixed, paraffin embedded human lung carcinoma specimens (45 cases of epidermoid carcinoma, 24 cases of small cell anaplastic carcinoma, 48 cases of adenocarcinoma, and 16 cases of large cell anaplastic carcinoma) as well as 14 cases with epithelial or biphasic mesothelioma and 7 cases of pleuritis carcinomatosa were incubated with biotinylated, suitably modified neoglycoproteins or heparin for detecting the corresponding endogenous sugar receptors (lectins). Histochemical reaction was demonstrated by use of the avidin-biotin-method. Significant differences were obtained between the different markers used for each cell type as well as between the different cell types. For separating small cell anaplastic carcinoma from non small cell anaplastic carcinoma the markers carrying the histochemically crucial maltose, fucose, and mannose residues are useful, showing a positive reaction to only 4-8% of the small cell anaplastic carcinoma cases compared to 60-80% of the non small cell carcinoma cases. For separation of mesothelioma from pleuritis carcinomatosa a carrier, modified by attachment of N-acetylglucosamine moieties is the most useful marker being negative in all cases of the analyzed mesothelioma cases and being positive in 62-80% of the adenocarcinoma cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550918 TI - Post-operative respiratory failure due to bilateral phrenic nerve palsy. AB - A 65 year old female patient developed a large left pleural effusion and a sternal split dehiscence following aorto-coronary artery bypass grafting. A second operation was performed to investigate and drain the pleural effusion and to repair the sternum. Subsequent to this operation the patient was in acute respiratory failure due to bilateral phrenic palsy. It is probable that the left phrenic nerve was damaged in the initial operation and the right nerve in the second. The patient's subsequent progress is described. PMID- 2550919 TI - Brachial plexitis complicating disseminated gonococcal infection. AB - A patient with brachial plexitis associated with disseminated gonococcal infection is presented. This is, to the best of our knowledge, the first case to be reported of this association. PMID- 2550920 TI - Mode of action of calcium antagonists: voltage-dependence and kinetics of drug receptor interaction. PMID- 2550921 TI - Deuterium isotope effects as a tool in the study of ethanol oxidation in rat liver microsomes. AB - The apparent kinetic deuterium isotope effect (I) on the oxidation of ethanol to acetaldehyde by washed rat liver microsomes was measured with (1-R)-[1-2H2, 1 14C]-ethanol (I1) and [1-2H2, 2-14C]-ethanol (I2) as substrates by a competitive technique involving only measurements of radioactivity. The average values were for non-induced rats, I1 = 1.57 and I2 = 2.23. When these two substrates were used with stereospecific enzymes (alcohol dehydrogenase and catalase) a small secondary effect was observed, causing I2 to be about 10% higher than I1. With non-stereospecific systems I2 was much larger than I1, and the values were connected by a simple formula. This relation in combination with use of the inhibitors, sodium azide and thiourea, made it possible to calculate tentatively the contribution to microsomal ethanol oxidation of catalase, a non-identified stereospecific enzyme, and non-stereospecific catalytic systems, as well as the isotope effects of the latter two systems. Measurements were made in microsomes from normal, phenobarbital treated, and acetone treated rats. For the stereospecific component an isotope effect of 1.4-1.5 was calculated for all three groups. For the non-stereospecific enzyme in acetone treated rats a value of 4.0 was found. Both the other groups showed a value about 2.7. The activity of the non-stereospecific system was about twice the normal in barbiturate treated, and 3 times the normal in the acetone treated group, where it contributed 70% of the total activity. The isotope effects on the changes in ethanol oxidation (the 'differential isotope effect') caused by inhibitors and activators were utilized to decide whether inhibitors were specific for a single reaction. Thus azide while inhibiting catalase completely, also inhibited other reactions. The large increase (5-6 times) in rate caused by Fe-ESDTA has an I2 of 1.6, equal to that for oxidation of ethanol by hydroxyl radicals. PMID- 2550922 TI - Cardiotoxic effects and the influence on the beta-adrenoceptor function of doxorubicin (Adriamycin) in the rat. AB - The possible relationship between the effect of the anthracycline-cytostatic doxorubicin (Dox) on the cardiac beta-adrenoceptor function in vitro and the development of delayed cardiotoxicity in vivo has been investigated in the rat. Dox (10(-5)-10(-4) M) blocked the chronotropic effect of isoprenaline on isolated atria in a competitive manner. Treatment with a single dose of Dox 5 mg/kg intravenously caused marked ECG changes manifested by progressive prologations of the Q alpha T and S alpha T-intervals, which amounted to 37% and 58% respectivity 5 weeks after the medication. At this time no beta-blocking action was detectable when tested on the isolated atria in the same rats. The results indicate that the delayed cardiotoxicity induced by Dox is not mediated by an interference with the cardiac beta-adrenoceptor function. PMID- 2550924 TI - Partial amino acid sequence of fructose-1,6-bisphosphatase from the blue-green algae Synechococcus leopoliensis. AB - Purified fructose-1,6-bisphosphatase from the cyanobacterium Synechococcus leopoliensis was S-carboxymethylated and cleaved with trypsin. The resulting peptides were purified by reversed-phase high performance liquid chromatography and the amino acid sequence of six of the purified peptides was determined by gas phase microsequencing. The results revealed sequence homology with other fructose 1,6-bisphosphatases. The obtained sequence data provides information required for the design of oligonucleotide hybridization probes to screen existing libraries of cyanobacterial DNA. The determination of the amino acid sequence of cyanobacterial proteins may yield important information with respect to the endosymbiotic theory of evolution. PMID- 2550923 TI - Oestrogen synthetase (aromatase) and hormone secretion in primary cultures of human placental trophoblast cells. Effects of cyclic AMP addition at the start of culture in attached and unattached cell populations. AB - Term placental trophoblast cells, released by trypsin digestion of placental villi, purified on a Percoll gradient and grown in serum-containing medium, differentiate within 24 to 48 h in culture from mononucleated cytotrophoblast like cells at the start of culture to highly multinucleated giant (syncytiotrophoblast-like) cells that are more active in hormonogenesis. To determine the changes in hormone biosynthesis and secretion that occur early in the trophoblast differentiation process in vitro, freshly isolated placental cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) containing 20 per cent FBS, in the presence and absence of cAMP for up to 48 h. Cell attachment and growth, oestrogen synthetase (aromatase) activity in attached and unattached cells, and secretion of human chorionic gonadotropin (hCG) and progesterone were studied. The aromatase specific activity, low in freshly isolated cells, increased fourfold in attached cells by 3 h, and achieved a 10- to 15-fold increase by 40 to 48 h. In attached cells grown with cAMP, aromatase activity was further stimulated by about fourfold, relative to the control. The aromatase activity of the unattached cells removed from the culture dishes at various times up to 48 h showed a biphasic response: the activity decreased by 18 h and then increased back to the fresh cell levels. The effect of cAMP on aromatase in these unattached cells was manifested by a two-fold stimulation of activity by 18 h, relative to control unattached cells. Secretion of hCG from both attached and unattached cells remained at a low level (less than 200 ng/mg protein) in control cells; in the presence of cAMP, hCG secretion was stimulated by tenfold after 40 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2550925 TI - Site specificity of the inhibitory effects of oligo(nucleoside methylphosphonate)s complementary to the acceptor splice junction of herpes simplex virus type 1 immediate early mRNA 4. AB - Oligo(nucleoside methylphosphonate)s complementary to the splice junction of herpes simplex virus type 1 immediate early pre-mRNAs 4 and 5 caused specific inhibition of herpes simplex virus type 1 growth. The dodecamer d(TpTCCTCCTGCGG) (deoxynucleoside methylphosphonate residues in italic) caused 50% and 98% decreases in herpes simplex virus type 1 titers at concentrations of 15 microM and 100 microM, respectively. d(TpTCCTCCTGCGG) inhibited viral but not cellular protein synthesis and decreased splicing of immediate early pre-mRNAs 4 and 5. Inhibition was highly sequence specific. A psoralen derivative of d(TpTCCTCCTGCGG) that can covalently bind to complementary sequences after exposure to 365-nm irradiation caused 90-98% inhibition of virus growth in cells treated with oligomer (5 microM) and irradiated at 1-3 hr postinfection. The data suggest that oligo(nucleoside methylphosphonate)s of appropriate sequence and derivatization may be effective as antiviral agents. PMID- 2550926 TI - Evidence that pp42, a major tyrosine kinase target protein, is a mitogen activated serine/threonine protein kinase. AB - pp42, a low-abundance 42-kDa protein, becomes transiently phosphorylated on tyrosine after stimulation of fibroblasts by a variety of mitogens, including epidermal growth factor, platelet-derived growth factor, phorbol 12-myristate 13 acetate, thrombin, and insulin-like growth factor II. The induction of pp42 phosphorylation on tyrosine by such diverse mitogenic agents suggests an important role for pp42 in the cascade of events necessary for cell transition from G0 into the cell cycle. However, as with most proteins identified on the basis of their tyrosine phosphorylation, the function of pp42 in cellular regulation is unknown. In this manuscript we report evidence that suggests that pp42 is a serine/threonine-specific protein kinase. Stimulation of 3T3-L1 cells with insulin has been shown to activate a cytosolic serine/threonine kinase capable of phosphorylating microtubule-associated protein 2 (MAP-2) and ribosomal protein S6 kinase II. This cytosolic serine/threonine protein kinase, which itself is phosphorylated on tyrosine, has been termed "MAP kinase". We now report that pp42 phosphorylation and MAP kinase activation occur in fibroblasts in response to similar mitogens, that the two proteins comigrate on one- and two dimensional polyacrylamide gels, and that the two proteins copurify chromatographically. The major peptides generated from purified MAP kinase by V8 protease digestion are present as a subset of the peptides in digests of pp42 excised from two-dimensional gels. Thus, the results suggest that MAP kinase is tyrosine-phosphorylated pp42. PMID- 2550927 TI - Evidence that v-src and v-fps gene products use a protein kinase C-mediated pathway to induce expression of a transformation-related gene. AB - Induction of the transformation-related gene 9E3 by the v-src and v-fps gene products (v-Src and v-Fps) is blocked in chicken embryo fibroblasts depleted of protein kinase C (PKC). PKC agonists induce 9E3 gene expression. Protein kinase inhibitors block v-Src- and v-Fps-induced 9E3 gene expression with the same dose response curves seen for PKC agonist-induced 9E3 gene expression. These data suggest that v-Src and v-Fps use a PKC-mediated signal-transduction pathway to induce expression of the transformation-related 9E3 gene. Consistent with this hypothesis, we find that both v-Src and v-Fps rapidly induce phosphorylation of a 67-kDa PKC substrate. PMID- 2550928 TI - Reciprocal inhibition of binding between interleukin 3 and granulocyte-macrophage colony-stimulating factor to human eosinophils. AB - 125I-labeled recombinant human interleukin 3 (IL-3) bound, at 4 degrees C, to a single class of high-affinity receptors on human eosinophils with an apparent dissociation constant (Kd) of 470 pM, but it did not bind to human neutrophils. 125I-labeled recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) also bound to a single class of high-affinity receptors on eosinophils with an apparent Kd of 44 pM and on neutrophils with an apparent Kd of 70 pM. These binding characteristics were consistent with the biological activities of IL-3 and GM-CSF on eosinophils and with the lack of stimulation of neutrophil function by IL-3. Specificity studies under conditions shown to prevent receptor internalization showed that the binding of 125I-labeled IL-3 to eosinophils was partially inhibited by GM-CSF but not by other cytokines. Reciprocal experiments with 125I-labeled GM-CSF showed that IL-3 but not other cytokines partially inhibited binding to eosinophils. In contrast, the binding of 125I-labeled GM-CSF to neutrophils was not inhibited by IL-3 or other cytokines tested. Quantitative inhibition binding experiments on eosinophils showed that the reciprocal inhibition between IL-3 and GM-CSF was not complete up to a concentration of heterologous ligand of 100 nM. These results show that (i) IL-3 binds to eosinophils but not neutrophils and (ii) IL-3 and GM-CSF specifically interact on the surface of eosinophils, providing a possible mechanism for the overlapping activities of IL-3 and GM-CSF on these cells. PMID- 2550929 TI - Disease tropism of c-erbB: effects of carboxyl-terminal tyrosine and internal mutations on tissue-specific transformation. AB - Avian leukosis virus induces erythroleukemia in chickens by proviral insertional mutation of the protooncogene c-erbB. The product of the insertionally activated c-erbB locus lacks the extracellular ligand-binding domain and is strictly leukemogenic. It has previously been demonstrated that the disease spectrum associated with aberrant c-erbB expression can be expanded by structural perturbation of the cytoplasmic domain of this protein. In this report, we use mutagenesis and retroviral vectors to identify specific mutations in the carboxyl terminal domain of the insertionally activated c-erbB product that are sufficient to activate the sarcomagenic potential of this protein. Interestingly, a point mutation in the kinase domain appears to be sufficient for sarcomagenic activation. However, removal of the terminal tyrosine residue of the c-erbB product, implicated in modulating kinase activity, does not lead to a fully transforming phenotype. These studies suggest that there are multiple ways to activate the fibroblast-transforming potential of the insertionally activated c erbB product. The conformation of this protein may play a more significant role in oncogenic activation than the phosphorylation status of the putative carboxyl terminal autophosphorylation site. PMID- 2550930 TI - cGMP-induced differentiation of the promyelocytic cell line HL-60. AB - cGMP is a second messenger that mediates numerous metabolic events; in the present work a role in myeloid cell differentiation was demonstrated. Nitroprusside and NaNO2, which activate cytosolic guanylate cyclase and increase the intracellular cGMP concentration, induced granulocytic differentiation of the human promyelocytic cell line HL-60; differentiation was measured by acquisition of the OKM1 antigen, morphological changes, and nitroblue tetrazolium reduction. When theophylline, a phosphodiesterase inhibitor, which by itself induced modest differentiation, was added to nitroprusside or NaNO2, differentiation increased in an additive fashion. The degree of differentiation correlated with the increase in the intracellular cGMP concentration. 8-Bromoguanosine 3',5'-cyclic monophosphate, a membrane-permeable cGMP analogue, also induced differentiation of HL-60 cells but was much more effective in the presence of theophylline, with the two agents interacting synergistically. The effect of theophylline in these studies could not be attributed to increasing the intracellular cAMP concentration. Dimethyl sulfoxide, and established inducer of differentiation of HL-60 cells, markedly enhanced the differentiation induced by nitroprusside and NaNO2. PMID- 2550931 TI - Transcription of thrombomodulin mRNA in mouse hemangioma cells is increased by cycloheximide and thrombin. AB - We have measured mRNA levels for thrombomodulin, an endothelial membrane cofactor for the activation of protein C by thrombin, in a mouse hemangioma cell line. Cycloheximide, an inhibitor of protein synthesis, increased levels of thrombomodulin mRNA, as measured in an S1 nuclease protection assay, to 2.5-4.0 times control levels. Thrombomodulin transcription in response to cycloheximide treatment, as determined by nuclear run-on analysis, was 3.9 +/- 1.3 (mean +/- SD) times that found in untreated cells. Thrombin also increased thrombomodulin mRNA levels to 151 +/- 21% (mean +/- SD) of control levels after 2 hr. Transcription increased in response to thrombin by 2.1- to 7.3-fold. The combination of thrombin and cycloheximide had no additive effect on thrombomodulin mRNA levels. Thrombin treatment of hemangioma cells also caused an increase in thrombomodulin protein synthesis to 142 +/- 17% (mean +/- SD) of control levels as determined by immunoprecipitation of [32S]methionine-labeled thrombomodulin. We conclude that thrombomodulin expression is determined in part by the rate of transcription and that thrombomodulin mRNA levels in hemangioma cells are increased by treatment with cycloheximide or thrombin. The increased transcription in response to cycloheximide suggests the existence of a labile protein repressor of thrombomodulin transcription. PMID- 2550932 TI - High levels of a parathyroid hormone-like protein in milk. AB - Expression of a parathyroid hormone-like protein (PLP), which is associated with hypercalcemia in malignancy, has recently been localized to normal lactating mammary tissue. We examined the possibility of an extramammary role of PLP by measuring its levels in serum and milk of lactating women. The levels of PLP by radioimmunoassay in serum of lactating and nonlactating women were indistinguishable [4.2 +/- 1.8 and 3.6 +/- 1.0 pg equivalents (eq) of PLP-(1-34) amide per ml, respectively]. As PLP was undetectable in some serum samples, this result does not exclude the possibility that lactation results in a small increase in serum levels of PLP. In contrast, high concentrations of immunoreactive PLP [40,000-75,000 pg eq of PLP-(1-34) amide per ml] and correspondingly high concentrations of bioactive PLP were found in human, rat, and bovine milk. A variety of processed bovine milk products had a PLP content similar to that of fresh bovine milk, whereas infant formulas had lower concentrations, ranging down to undetectable. Although the physiological role of PLP in lactation is unknown, the data establish the presence of PLP in milk and suggest the possibility that PLP may be important in neonatal calcium homeostasis. PMID- 2550933 TI - Cloning of the cDNA and functional expression of the 47-kilodalton cytosolic component of human neutrophil respiratory burst oxidase. AB - Neutrophil NADPH oxidase is a multicomponent enzyme that is activated to generate superoxide anion and is defective in the cells of patients with chronic granulomatous disease. It requires both membrane and cytosolic components, the latter including 47- and 67-kDa proteins recognized by the polyclonal antiserum B 1. Immunoscreening of an induced HL-60 lambda ZAP cDNA library yielded seven cross-hybridizing cDNAs encoding the 47-kDa component. Fusion proteins of 22-50 kDa were recognized by B-1. Antiserum against a fusion protein recognized a 47 kDa protein in normal neutrophils but not in those from patients with autosomal chronic granulomatous disease who lack the 47-kDa cytosolic oxidase component. In a cell-free NADPH oxidase system full-length and C-terminal fusion proteins augmented superoxide generation and reconstituted the cytosolic defect of a patient missing the 47-kDa protein. The cDNA hybridized with a 1.4-kilobase mRNA from induced HL-60 cells. The longest cDNA contained an open reading frame encoding a protein of 41,440 Da with a calculated pI of 10.4, an N-terminal glycine, sites favorable for phosphorylation, a nucleotide binding domain, and a region of homology to the src protein kinases, phospholipase C, and alpha-fodrin. These structural features are pertinent to proposed functional roles of the protein in the respiratory burst oxidase. PMID- 2550934 TI - Loss of heterozygosity on chromosome 1q in human breast cancer. AB - Cytogenetic markers involving the long arm of chromosome 1 are the most frequently observed karyotypic changes seen in breast cancer. Based on cytogenetic data, we have used polymorphic DNA markers to search for allelic losses at this chromosome region among 48 breast carcinomas. For SPTA1, allelic losses were seen in 6 of 26 (23%) informative carcinomas, while 3 of 13 (23%) and 5 of 19 (26%) informative patients showed losses at AT3 and D1S53, respectively. The background frequency of allelic loss was obtained from data using 3 other loci on the 1q arm and 2 on the p arm of chromosome 1. With these markers, only 6 of 62 informative patients (8%) showed an allelic loss, with the range being 0 13%. The allelic losses seen on 1q, which were found in 9 carcinomas, comprised an overlapping set; the common region deleted was approximately 26 centimorgans on the q arm of chromosome 1 (bands q23-32 between AT3 and D1S53). These results suggest that inactivation of a gene(s) located on 1q23-32 might contribute to the genesis of breast cancer. PMID- 2550935 TI - Protection of macaques against simian AIDS by immunization with a recombinant vaccinia virus expressing the envelope glycoproteins of simian type D retrovirus. AB - Simian AIDS (SAIDS) is an endemic disease of macaques that shares many characteristics with AIDS in humans. SAIDS is etiologically linked to infection by a type D retrovirus, SAIDS retrovirus (SRV). Immunization with an inactivated whole-virus vaccine was shown to protect macaques against infection by SRV serotype 1. To identify the antigen(s) responsible for eliciting protective immunity, we have constructed a recombinant vaccinia virus (v-senv5) that expresses the envelope glycoproteins of SRV serotype 2 (SRV-2/W). Pig-tailed macaques (Macaca nemestrina) immunized with v-senv5 showed lymphoproliferative responses to purified SRV-2/W. They also generated antibodies that neutralized SRV-2/W infectivity in vitro and mediated antibody-dependent cellular cytotoxicity against SRV-2-infected cells. Four v-senv5-immunized animals, together with four control animals, were challenged intravenously with 5 x 10(3) tissue culture infectious doses of SRV-2/W. As early as 2 weeks after challenge, three of four control animals became viremic, and two of these three animals also seroconverted. The animal that was viremic but remained antibody negative died of symptoms of SRV infection 6 1/2 weeks after challenge. In contrast, all four v senv5-immunized animals remained healthy, virus-free, and seropositive against only the immunizing envelope antigens. These results indicate that immunization with a recombinant vaccinia virus expressing the envelope antigens of SRV-2/W protects primates from infection by a retrovirus that causes immunodeficiency diseases. PMID- 2550936 TI - T-type calcium channels mediate the transition between tonic and phasic firing in thalamic neurons. AB - Thalamic neurons undergo a shift from tonic to phasic (burst) firing upon hyperpolarization. This state transition results from deinactivation of a regenerative depolarizing event referred to as the low-threshold spike. Isolated adult guinea pig thalamic (dorsal lateral geniculate) neurons exhibited low threshold spikes that could be blocked by low concentrations of nickel but were unaffected by the dihydropyridine nimodipine. Whole-cell voltage-clamp recordings from these cells demonstrated a low-threshold, rapidly inactivating (T) Ca2+ current that manifested similar voltage dependency and time course as the low threshold spike. Like low-threshold spikes, the T-type Ca2+ current was eliminated by nickel but was unaffected by nimodipine. In thalamic neurons, T type Ca2+ channels underlie the low-threshold spike and, therefore, play a critical role in regulating the firing pattern of these cells. PMID- 2550937 TI - Single-channel recording in myelinated nerve fibers reveals one type of Na channel but different K channels. AB - Amphibian myelinated nerve fibers were treated with collagenase and protease. Axons with retraction of the myelin sheath were patch-clamped in the nodal and paranodal region. One type of Na channel was found. It has a single-channel conductance of 11 pS (15 degrees C) and is blocked by tetrodotoxin. Averaged events show the typical activation and inactivation kinetics of macroscopic Na current. Three potential-dependent K channels were identified (I, F, and S channel). The I channel, being the most frequent type, has a single-channel conductance of 23 pS (inward current, 105 mM K on both sides of the membrane), activates between -60 and -30 mV, deactivates with intermediate kinetics, and is sensitive to dendrotoxin. The F channel has a conductance of 30 pS, activates between -40 and 60 mV, and deactivates with fast kinetics. The former inactivates within tens of seconds; the latter inactivates within seconds. The third type, the S channel, has a conductance of 7 pS and deactivates slowly. All three channels can be blocked by external tetraethylammonium chloride. We suggest that these distinct K channel types form the basis for the different components of macroscopic K current described previously. PMID- 2550938 TI - Spontaneous electrical activity of interstitial cells of Cajal isolated from canine proximal colon. AB - Interstitial cells of Cajal (ICC) have been suggested as pacemaker cells in the gastrointestinal tract. A method was developed to isolate ICC from the slow-wave pacemaker region of the canine proximal colon. These cells were identified under phase-contrast microscopy, and their identity was verified by comparing their ultrastructure with the morphology of ICC in situ. Patch-clamp experiments demonstrated that these cells are excitable; voltage-dependent inward and outward currents were elicited by depolarization. Inward current transients were identified as calcium currents. A portion of the outward current appears to be due to Ca2+-activated K channels commonly expressed in these cells. ICC were also spontaneously active, generating electrical depolarizations similar in waveform to slow-wave events of intact colonic muscles. These findings are consistent with the hypothesis that ICC initiate rhythmicity in the colon. PMID- 2550940 TI - In vitro evaluation of a novel, non-competitive opioid receptor antagonist. PMID- 2550939 TI - Functional expression of the amiloride-sensitive sodium channel in Xenopus oocytes. AB - Expression of the amiloride-sensitive sodium channel was examined in Xenopus oocytes that were microinjected with A6 cell mRNA. Amiloride-inhibitable 22Na flux could be measured in intact oocytes 2-3 days after injection with 25 ng of poly(A)+ RNA isolated from aldosterone-treated A6 cells. The rate of 22Na uptake was approximately 15-fold greater in oocytes microinjected with 25 ng of poly(A)+ RNA than in water-injected control oocytes. An increase in 22Na uptake by mRNA injected oocytes occurred whether the mRNA was isolated from A6 cells grown on a porous or nonporous support. In the presence of 4 mM NaCl, amiloride caused dose dependent inhibition of 22Na uptake in mRNA-injected oocytes, which was half maximal at 6 x 10(-8) M. Both 1 microM amiloride and 0.1 microM benzamil inhibited 22Na uptake in mRNA-injected oocytes by greater than 95%, whereas less than 50% inhibition occurred with 1 microM 5-(N-ethyl-N-isopropyl)amiloride. When A6 cell mRNA was size fractionated by sucrose density-gradient centrifugation, amiloride-sensitive 22Na uptake was expressed predominantly by oocytes injected with mRNA from two contiguous fractions. PMID- 2550941 TI - A comparison among antinociceptive bioassays for mu and delta agonists. PMID- 2550942 TI - RX809055AX in vivo: a long lasting opioid antagonist at mu and delta receptors. PMID- 2550943 TI - In vitro comparison of efficacies of "full" kappa opioid agonists. PMID- 2550945 TI - Inhibition of gastric H+, K+ adenosine triphosphatase (ATPase) by substituted naphthylalkylimidazoles. PMID- 2550944 TI - Comparison of PD 117302, a kappa agonist, and morphine on colonic transit and formalin-induced pain in rats. PMID- 2550946 TI - The degree of atrial natriuretic peptide (ANP)-induced contraction inhibition is dependent on spare receptors. PMID- 2550947 TI - The treatment of Wilms' tumor: an update. PMID- 2550948 TI - Recent advances in the surgical treatment of bilateral Wilms' tumor. AB - Definite progress has been made in the treatment of bilateral Wilms' tumor with marked improvement in the prognosis. Past aggressive surgical therapy has been recently replaced by a more conservative approach. Double or triple drug chemotherapy combined with conservative nephron sparing surgery is indicated for tumors with "favorable" histology. More aggressive ablative surgical therapy combined with chemotherapy and low dose radiation therapy are reserved for tumors with "unfavorable" histology. Fine needle aspiration biopsy, by both smears and cell block, combining light and electron microscopy approach, is an accurate method in establishing the diagnosis. It has some limitation in the precise histologic differentiation between "favorable and unfavorable" tumors. Treatment should be individualized according to clinical, histologic and morphologic factors. A multispecialty team effort undoubtedly, constitutes the most important element in the successful treatment of these tumors. PMID- 2550949 TI - Phase II trial with etoposide (VP16) plus ifosfamide plus high-dose cisplatin (VIhP regimen) in refractory germ cell tumors. PMID- 2550950 TI - Third-line chemotherapy of resistant advanced testicular cancer. AB - The efficacy of combined chemotherapy of Vepesid + Holoxan +/- Adriblastin as third-choice was studied in advanced testicular cancer patients refractory to or recurrent after first- and second-line cytostatic therapy. Between September 1981 and January 1988 49 evaluable patients were treated with Vepesid (VP-16213 - 100 mg/m2 days 1-5), Holoxan (40 mg/kg days 1-5), hydration, urine alkylation + Uromitexan +/- Adriblastin (40 mg/m2 day 1). The single dose of Uromitexan was 20% of the daily dose of Holoxan, and the patients received it i.v. just prior to Holoxan administration (hr 0), then 4 and 8 hrs later. Two patients got into CR and 10 to PR. The rate of remission was 24.48%. The most severe side effect was leukopenia. The elevation of CN and Se creatinine was transient and mild. In those cases where Holoxan was not included in the first or second-line regimens, when combined with Vepesid and Adriblastin as third-choice therapy one could achieve further improvement. In case of CR the prolongation of life is also noteworthy. PMID- 2550951 TI - Prevention of bilateral testicular cancer: significance of detection and treatment of carcinoma in situ. PMID- 2550952 TI - Macrophage receptor-mediated processing and regulation of advanced glycosylation endproduct (AGE)-modified proteins: role in diabetes and aging. AB - Tissue and cell surface proteins modified nonenzymatically by glucose are shown to be removed by macrophages through a recently characterized high affinity receptor. Insulin appears to be a potent suppressor of this macrophage AGE removal activity, while TNF acts as a stimulant. Coupling of AGE-proteins to their AGE-receptor results in TNF and IL-1 synthesis and secretion. This suggests that AGE may act as a signal for growth-promoting factor secretion in a coordinated replacement process during tissue remodeling. A greater than 2-fold decrease in receptor number and binding capacity found in cells from aged mice as compared to young suggests that aging may adversely affect the AGE-receptor efficiency and by impeding crosslinked AGE-protein removal add to ongoing aging tissue damage. PMID- 2550953 TI - Protection against the consequences of intravascular coagulation by reticuloendothelial stimulation. PMID- 2550954 TI - Long term administration of dopamine: is there a development of tolerance? PMID- 2550955 TI - Effects of bacterial exo- and endotoxins on endothelial arachidonate metabolism. PMID- 2550956 TI - Lipidperoxidation in a canine model of hypovolemic-traumatic shock. PMID- 2550957 TI - Modulation of resynthesis of 1-alkyl-2-arachidonyl-glycero-3-phosphocholine and phosphatidylinositols for interception in vivo of free arachidonic acid, lyso PAF, diacyl-glycerols, and phosphoinositides. AB - Thermal injury causes directly a liberation of inositolphosphates, diacylglycerols, free arachidonic acid, and lyso PAF from eukaryotic cells. From lyso PAF derivates PAF, from free arachidonic acid are derivating PG, LT, and TX. These "soluble mediators" are stimulating inflammatory cell populations in a feedback mechanism: the stimulus activates the inflammatory cells to produce the same soluble mediators (Fig.1). The arising soluble mediators are the take off for the inflammation cascade causing as later step the activation of kinin, clotting, and complement systems. The pure biochemical lesions at the onset results in the clinical manifestation of oedema, increased dermal temperature, and pains. The possibilities for prevention and allevation of early pain, due to the acute burn, lie in the inhibition of the spreading out of inflammatory mediators (Bauer 1987a) (Fig.2). PMID- 2550958 TI - TNF-induced organ changes in a chronic ovine model--possible role of leukocytes. PMID- 2550959 TI - Patterns of endocrine secretion during sepsis. AB - In septic patients the clinical course of the disease is characterized by high DIT and rT3 serum concentrations as well as a low T3-syndrome. While rT3 is elevated in almost all critically ill patients, the increase in DIT is indicative of severe infection. Prolactin levels are regularly elevated in sepsis although to variable degrees. Catecholamines and vasopressin should be regarded as acute responders. The pattern of cortisol secretion is uncertain. In most situations the secretion appears to be elevated; the circadian rhythm is disturbed. PMID- 2550960 TI - Infections in thalassemic patients (hepatitis and bone marrow transplantation related infections excluded). PMID- 2550961 TI - CMV infections in thalassemia patients after BMT. PMID- 2550962 TI - Complications of blood transfusion in thalassemia. PMID- 2550964 TI - Localization and characterization of prostaglandin E1 receptors in rat small intestine. AB - While prostaglandins of the E series are known to affect several small intestinal functions, their cellular mechanisms are poorly understood. The purposes of our study were to determine whether receptors for PGE are present in rat small intestine and to locate and characterize the receptor binding in the subcellular fractions. Small intestinal binding of prostaglandin E1 was significantly higher than that of prostaglandin E2. Highest receptor binding for prostaglandin E1 was found in the plasma membrane fraction of isolated small intestinal enterocytes. Curvilinearity of prostaglandin E1 binding in plasma membranes upon Scatchard analysis indicated two receptor binding sites in rat small intestine. Competitive binding studies demonstrated that receptor binding was highest for prostaglandins of the E series. These studies are the first to demonstrate specific prostaglandin E1 receptors in different subcellular fractions of rat small intestine. We suggest that receptor binding of prostaglandin E may be an important initial step in the mechanism of prostaglandin-E-induced responses in the small intestine. PMID- 2550963 TI - Release of leukotriene B4 from rat Kupffer cells. AB - In order to examine the production of leukotriene B4 (LTB4) from Kupffer cells, Kupffer cells isolated from the normal rat liver were incubated with calcium ionophore A23187, opsonized zymosan, or platelet activating factor (PAF), and the amount of LTB4 in the culture supernatant was determined by the combined technique of reverse-phase high-performance liquid chromatography and radioimmunoassay. As a result, when activated in vitro with calcium ionophore A23187, Kupffer cells generated LTB4. When Kupffer cells were stimulated with calcium ionophore after 10-min preincubation with AA861, a selective 5 lipoxygenase inhibitor, the release of LTB4 from Kupffer cells was markedly suppressed. PAF, which is a phospholipid mediator having a wide spectrum of biological activities, significantly enhanced the release of LTB4 from Kupffer cells stimulated with calcium ionophore or opsonized zymosan. Even when the Kupffer cell were not stimulated with calcium ionophore or opsonized zymosan, LTB4 production was significantly increased by PAF. Thus, our studies indicate that Kupffer cells could generate LTB4 as well as polymorphonuclear leukocytes and macrophages. In addition, it is suggested that Kupffer cells may be able to modify inflammatory and immunological events in the liver tissue by the release of LTB4. PMID- 2550966 TI - Metabolism of LTC4 to LTD4 and LTE4 in isolated guinea pig lung lobes. AB - Leukotrienes are known to be easily metabolized to other substances. But the metabolic fates of LTC4 and LTD4 have not been established in the intact lung. In this investigation we perfused isolated guinea pig lung lobes and injected synthesized LTC4 and LTD4. The effluent was assayed by HPLC. LTD4 and LTE4 were detected following perfusion of LTC4, and LTE4 was detected following perfusion of LTD4. These results suggest that perfused guinea pig lung lobes may metabolize LTC4 to LTD4 and LTE4, and LTD4 to LTE4. PMID- 2550965 TI - Distribution and metabolism of leukotriene C4 after cisternal injection in guinea pigs. AB - Following cisternal injection of [3H8]LTC4 into guinea pigs, leukotriene metabolites were identified in the brain, cerebellum, perilymph, blood, liver and kidneys. LTC4 was metabolized into LTD4 and LTE4 in the cerebrospinal fluid and LTE4 was transported into the blood for general circulation and uptake into the liver and kidneys. The excretion of LTE4 from CNS to blood seemed to be the rate limiting step in the elimination of leukotrienes from the body. Leukotrienes were also transported into the perilymph. The conversion of LTC4 into LTD4 and LTE4 was lower in perilymph as compared to the cerebrospinal fluid, suggesting a rate limiting function of the cochlear aqueduct that can be defined as a cerebrospinal fluid-labyrinth barrier. PMID- 2550967 TI - Stimulation of anaphylaxis in the mouse footpad by dietary fish oil fatty acids. AB - The effect of fish oil-derived omega-3 (omega-3) fatty acids on anaphylaxis, Arthus and delayed type hypersensitivity reactions in mice has been investigated. Mice on a normal chow diet were fed eicosapentaenoic acid and docosahexaenoic acid at a dose of 500 and 333 mg/kg/day, respectively, by a gastric tube over a period of 61 days. Control groups were given water, safflower oil or oleic acid. Anaphylactic and Arthus type reactions were induced in the mouse footpad using bovine serum albumin as an antigen. Carrageenin was utilized to produce a delayed type hypersensitivity reaction. The animals fed omega-3 fatty acids induced a more anaphylactic foodpad reaction. There was no significant effect of the diet on Arthus and delayed type hypersensitivity responses. There was no effect of the fish oil-supplemented diet on production of antibodies to bovine serum albumin. Synthesis of prostaglandin E2 by peritoneal macrophages was significantly inhibited in the animals fed omega-3 fatty acid-enriched fish oil, while leukotriene B4 production was not affected. These results suggest that a diet enriched in omega-3 fatty acids modulates production of arachidonic acid metabolites and this may influence anaphylaxis, but not Arthus and cellular mediated hypersensitivity responses. PMID- 2550968 TI - Eicosanoid synthesis by alveolar macrophages in rats with malignant mammary tumors: differences in rats treated with and without carrageenan implants. AB - Eicosanoid synthesis by alveolar macrophages (AM), harvested from tumor bearing animals, was measured after tumor inoculation in rats treated with or without carrageenan (carra), an immunomodulating agent. After incubation of the cells with [14]C-arachidonic acid and the Ca-ionophore A23187, samples were measured by high pressure liquid chromatography (HPLC). From the HPLC profiles the lypoxygenase products, 12-hydroxyeicosatetraenoic acid (12-HETE), 15-HETE, and leukotriene-B4 (LTB4) were determined as well as the cyclooxygenase products, prostaglandin (PG)E2, PGF2 alpha and TXB2. After tumor inoculation AM-synthesis of lipoxygenase products tended to increase to values twice those of the base line values, whereas cyclooxygenase products showed subnormal values. In the non treated animals, 10 days after tumor inoculation, statistically significant increases in 12- and 15-HETE, LTB4 and PGE2 were observed when compared with carra treated animals. Later measurements did not show these differences in AM metabolism. AM metabolism was (negatively) correlated with the number of macrophages, which was particularly evident in the correlation with 12-HETE synthesis. PMID- 2550969 TI - Plasma fatty acid levels in patients with acquired immune deficiency syndrome and in controls. AB - Polyunsaturated fatty acids (PUFAs) are known to modulate the immune system in vivo and to inactivate envelope viruses in vitro. Patients with AIDS had low total plasma lipid levels and low levels of a number of individual fatty acids. However, the C20 and C22 essential fatty acids of the n-3 series were selectively and highly significantly reduced. Normalization of these fatty acid levels in AIDS patients may be a worthwhile therapeutic aim. PMID- 2550970 TI - 12(R)-hydroxyeicosatetraenoic acid is a neutrophil chemoattractant in the cavine, lapine, murine and canine dermis. AB - Psoriasis is a disease state characterized by epidermal proliferation, neutrophil infiltration, along with release of the proinflammatory mediators leukotriene B4(LTB4) and 12(R)-hydroxyeicosatetraenoic acid [12(R)-HETE]. LTB4 and 12(R)-HETE are chemoattractant to the neutrophil, the latter approximately 1000x less potent. LTB4 and 12(R)-HETE are present in psoriatic scale, the latter in quantities so much greater than LTB4 that it is proposed as a primary mediator of neutrophil infiltration in psoriasis. 12(R)-HETE, synthesized in optically pure form by a new, shorter route, was injected into the dermis of the cavine, lapine, canine, mouse and rat. At doses up to 50 mu gm per intradermal site, 12(R)-HETE was chemoattractant to the neutrophil (as assessed by dermal myeloperoxidase levels) with response in the cavine greater than canine greater than lapine greater than mouse greater than rat. PMID- 2550971 TI - Acute effects of unsaturated fatty acids in splanchnic artery occlusion shock. AB - Diets enriched with omega-3 unsaturated fatty acids are associated with decreased hypercholesterolemia and decreased risk of ischemic and atherosclerotic diseases. We studied the acute intravascular effects of some of these unsaturated fatty acids (i.e., eicosapentaenoic acid, EPA; docosahexaenoic acid, DHA) along with omega-6 unsaturated fatty acids, (i.e., linoleic and linolenic acid) in splanchnic artery occlusion (SAO) shock in rats. Anesthetized rats subjected to total occlusion of the celiac and superior mesenteric arteries for 40 minutes followed by reperfusion usually resulted in a fatal outcome 90-120 minutes after releasing the clamps. SAO shock rats treated with the omega-3 unsaturated fatty acid, EPA, exhibited an improved survival time and rate (p less than 0.05 from vehicle) compared to those receiving only vehicle (i.e., 50% ethanol). EPA and DHA treated SAO rats also exhibited lower plasma activities of the lysosomal protease, cathepsin D, free amino-nitrogen compounds, and the cardiotoxic peptide, myocardial depressant factor. These results indicate that omega-3 unsaturated fatty acids, especially EPA, have some acute beneficial effects in SAO shock in rats. PMID- 2550972 TI - Effects of scopolamine on extracellular acetylcholine and choline levels and on spontaneous motor activity in freely moving rats measured by brain dialysis. AB - The present study demonstrates the feasibility of measuring acetylcholine (ACh) and choline in perfusate samples collected by in vivo brain dialysis in the frontal cortex and hippocampus of freely moving rats in which spontaneous motor activity could be measured simultaneously. Systemically administered scopolamine increased the output of ACh about 10-fold and 20-fold in the frontal cortex and hippocampus, respectively. By contrast, scopolamine decreased the choline level in the extracellular fluid about 2-fold in both brain regions, possibly owing to enhanced choline uptake into the presynaptic nerve terminals. Scopolamine also increased spontaneous motor activity over the same time course as the changes in ACh and choline. These results indicate that the in vivo brain dialysis technique applied to freely moving rats may be useful in investigating ACh turnover and in studying the relation between cholinergic transmission and behavioral functions. PMID- 2550973 TI - The Lesion Game: a special communication. AB - The Lesion Game is a personal computer program that allows the user to view and study muscle innervations and nerve lesions of the brachial plexus. The core of the program is a table that consists of 44 brachial plexus lesions, 50 muscles, and 2 sensations (for brachium and forearm) of the upper extremity, which are innervated by the brachial plexus. After the program randomly selects a lesion, the user attempts to find the lesion in as few guesses (manual muscle tests) as possible. As muscles are selected (muscle tested), the computer searches the table to find the appropriate "strength" of weak or normal, based on the location of the randomly selected lesion. After displaying the strength of the muscle selected, a graphic representation of the strength is shown on a diagram of the brachial plexus. The graphic aspect of the program helps the user to visualize areas of the brachial plexus that may still contain the lesion. While playing the Lesion Game, the user can view a detailed picture of the brachial plexus, view all the possible lesions of the Lesion Game, or view charts of upper extremity innervations. An additional program mode allows beginning users to view and study muscle innervations without having to solve lesions. The program is extremely simple to use because it is entirely mouse-driven. PMID- 2550974 TI - Photosensitization of anticancer agents--8. One-electron reduction of mitoxantrone: an EPR and spectrophotometric study. AB - An efficient method of one-electron reduction of the anticancer agent mitoxantrone is described. The method depends on illumination of a suitable photosensitizer absorbing blue light [acriflavine, anthrapyrazole, or Ru(bpy)2+(3)] in the presence of the drug and an electron donor, such as NAD(P)H, in deaerated solutions. An EPR spectrum, assigned to a semiquinone of mitoxantrone, is generated under these conditions and identified by spectral simulation. Decay of this species, attributed to a radical-radical reaction, gives a second order rate constant of 1.7 x 10(2) M-1 s-1 in organic media [dimethylsulfoxide (DMSO)/pH 8 buffer, 1:1 vol/vol] but is more rapid (approximately 10(4) M-1 s-1) in aqueous media under comparable conditions. The considerably decreased lifetime of the mitoxantrone radical at pH 5 is attributed to an additional electron transfer, promoted by protonation of the radical, and/or to an accelerated recombination of neutral radicals, leading to an EPR silent species. Parallel spectrophotometric studies on the generation of the mitoxantrone reduced species by photosensitized reduction are described. The method offers convenient access to a key radical species involved in the metabolism and possible mode of action of this clinical anticancer agent. PMID- 2550975 TI - Cortical Na+,K+-ATPase of immature rats following bicuculline-induced seizures. AB - The Na+,K+-ATPase activity was investigated in cerebral cortex homogenates of 7-, 12- and 18-day-old rats in which seizures were induced by systemic (i.p.) administration of bicuculline. Na+,K+-ATPase activities in control animals increased during postnatal development, but they were not significantly influenced by seizure activity when determined under optimal conditions in vitro. Although the ratio of neuronal vs. non-neuronal cells in cortical samples of 7-, 12- and 18-day-old rats was different, there was a remarkable similarity in the activation curves for K+, obtained for Na+,K+-ATPase of all age groups under normal conditions; 50% of enzyme activities were attained at 1 mmol.l-1 K+ and the maximal activities were found around 10 mmol.l-1 K+. The activation curves for K+ in rats with bicuculline-induced seizures were not significantly different from those of the controls. PMID- 2550976 TI - Sex pheromones and plasmid transfer in Enterococcus faecalis. AB - Plasmid-free Enterococcus faecalis excrete peptides (sex pheromones) which specifically induce a mating response in strains harboring certain conjugative plasmids. The response is characterized by the synthesis of a "fuzzy" surface material, visible by electron microscopy, which is believed to facilitate the aggregation of donors and recipients. Transconjugants which receive a specific plasmid shut down the production of endogenous pheromone; however, they continue to produce pheromones specific for donors harboring different classes of plasmids. In this review, we summarize what is known about the biochemistry and genetics of this phenomenon. Some emphasis is given to the hemolysin plasmid pAD1 and the regulation of its conjugal transfer. PMID- 2550977 TI - Physical and biological characterization of linear DNA plasmids of the yeast Pichia inositovora. AB - Three cryptic DNA plasmids have been identified in a strain of the yeast Pichia inositovora that are 18, 13, and 10 kbp in size. All are sensitive to digestion by DNase I, restriction endonucleases, and exonuclease III, but are resistant to the activities of RNase A and lambda exonuclease. These results indicate that each plasmid is a linear DNA molecule whose 5' ends are protected. A restriction map has been developed for each of the plasmids, demonstrating that each is unique and confirming their linear nature. The plasmids are a major constituent of DNA prepared from whole cells, but are absent from DNA preparations of purified mitochondria and nuclei, indicating that the plasmids are located in the cytoplasm. These plasmids share many of the physical characteristics described for the linear plasmids of the yeasts Kluyveromyces lactis and Saccharomycopsis crataegensis. Unlike the linear plasmids of K. lactis, however, they appear not to be capable of killer toxin production. PMID- 2550978 TI - The EcoDXX1 restriction and modification system: cloning the genes and homology to type I restriction and modification systems. AB - The Escherichia coli plasmid pDXX1 codes for a type I restriction and modification system, EcoDXX1. A 15.5-kb BamHI fragment from pDXX1 has been cloned and contains the hsdR, hsdM, and hsdS genes that encode the EcoDXX1 system. The EcoDXX1 hsd genes can complement the gene products of the EcoR124 and EcoR124/3 hsd systems, but not those of EcoK and EcoB. Hybridization experiments using EcoDXX1 hsd genes as a probe demonstrate homology between EcoDXX1 and EcoR124 and EcoR124/3 restriction-modification systems, but weak or no homology between EcoDXX1 and EcoK or EcoB systems. PMID- 2550979 TI - Replication properties of mini-Rts1 derivatives deleted for DnaA boxes in the replication origin. AB - Mini-Rts1 was found to be unable to replicate in a dnaA-null mutant. However, a mini-Rts1 derivative lacking entire tandem DnaA boxes in the replication origin retained the replication ability in a dnaA+ host although its copy number was about half that of the mini-Rts1 having complete DnaA boxes. Mini-Rts1cop1 that contains a high copy number mutation in repA was found to replicate more efficiently than mini-Rts1 of wild repA when DnaA boxes were deleted. In addition, the copy number of mini-Rts1cop1 without DnaA boxes increased 1.5-fold upon removal of incI iterons, whereas that of mini-Rts1 without DnaA boxes did not increase after the iterons were deleted. These indicate that the RepAcop1 protein can initiate the replication of mini-Rts1 efficiently even when DnaA boxes are absent from the origin of replication. PMID- 2550980 TI - Intermolecular plasmid recombination in fibroblasts from humans with DNA damage processing defects. AB - We have evaluated the ability of immortalized human fibroblasts to recombine transfected plasmid DNA. A number of cell lines from normal individuals and from patients with DNA damage-processing defects were examined. Two plasmid recombination substrates were derived from pSV2neo and contained nonoverlapping deletions in the aminoglycoside phosphotransferase II gene. Intermolecular recombination was assessed by two methods after cotransfection. In a short-term, extrachromosomal recombination assay, low molecular weight DNA was extracted from the human cells 48 h after transfection, and recombinant plasmids were detected by transformation into appropriate indicator bacteria. In a long-term stable recombination assay the fibroblasts were cotransfected and G418-resistant colonies allowed to form. By the former assay all but two cultures were recombination-proficient, whereas all were recombination-proficient by the latter assay. The efficiency of transfection of human cells with plasmids appears to be a major variable affecting recombination. Recombination can be stimulated by uv irradiation of plasmid DNA prior to transfection. Cells from patients with Fanconi anemia, ataxia telangiectasia, and xeroderma pigmentosum complementation groups A, C, D, E, and G are not defective at intermolecular plasmid recombination. PMID- 2550981 TI - Structural analysis of Tc1 elements in Caenorhabditis elegans var. Bristol (strain N2). AB - The transposable element Tc1 in the genome of Caenorhabditis elegans var. Bristol strain N2 is very stable. In order to investigate possible causes of Tc1 immobility in this strain 17 individual isolates have been cloned and characterized with regard to their structure and genomic environment. Ten of 16 elements examined had identical restriction maps, and at least 1 of these (#7) showed a high level of somatic excision. Two of the elements had altered restriction sites, 2 had different internal deletions of about 700 bp, 1 had an 89-bp terminal deletion, and 1 a 54-bp insertion. When DNA sequences flanking the N2 Tc1 elements were used as probes in genomic hybridizations, it was found that most N2 elements are located in regions of repetitive DNA. Furthermore when hybridizations to DNA from N2 and var. Bergerac strain B0 were performed, a major band of the same size was observed in both strains. Two flanking sequences identified strain polymorphic sites hP2(IV) and hP3(IV). In at least one of these cases, a rearranged Tc1 was present in the B0 strain at the same location. The fact that all or most of the Tc1 elements are in the same location in N2 and B0 adds support to the hypothesis that the high copy number B0 strain arose from amplification of Tc1 copies in a N2-like strain. The N2 Tc1 elements are highly conserved; however, intact elements had fewer nucleotide changes than the rearranged elements. These results may indicate that the intact Tc1 elements in N2 are functionally active and subject to selective pressure. PMID- 2550982 TI - Cloning and characterization of variable-sized gypsy mobile elements in Drosophila melanogaster. AB - A cosmid genomic library from a known gypsy-induced forked mutation, f1, was screened by 32P-labeled gypsy transposable element. Of more than 250 positive clones we randomly selected 21 for in situ hybridization to wild-type polytene chromosomes. Two clones hybridized to region 15F on the X-chromosome, the cytological position of forked. A third clone hybridized to at least 17 sites on the chromosomes indicating the presence of repetitive sequences in the gypsy flanking DNA. All clones labeled the centromeric regions heavily. Ten clones, including the two hybridizing at 15F, were chosen for further analysis, and restriction mapping allowed us to place them into three groups: (1) full-length, (2) slightly diverging, and (3) highly diverging gypsy elements. Group (2) is missing the XbaI site in both their long terminal repeats (LTRs) as well as the middle HindIII site; four of these gypsy elements also have a approximately 100 bp deletion at the 5' LTR. The group (3) gypsy transposons are missing one LTR and also have highly diverging DNA sequences. The restriction analyses further imply that most of these different gypsy elements are present in more than one copy in the genome of the f1 stock used in this study. The results raise intriguing questions regarding the significance of transposable elements in evolution and biological functions. PMID- 2550983 TI - Nucleotide sequence of a novel kanamycin resistance gene, aphA-7, from Campylobacter jejuni and comparison to other kanamycin phosphotransferase genes. AB - A novel kanamycin phosphotransferase gene, aphA-7, was cloned from a 14-kb plasmid obtained from a strain of Campylobacter jejuni and the nucleotide sequence of the gene was determined. The presumed open reading frame of the aphA 7 structural gene was 753 bp in length and encoded a protein of 251 amino acids with a calculated weight of 29,691 Da. A 29-kDa protein was demonstrated in Escherichia coli maxicells containing the cloned aphA-7 gene. A ribosomal binding site corresponding to 5 of 8 bases of the 3' end of the E. coli 16S rRNA was 8 bp upstream of the start codon. Sequences corresponding to the -35 and -10 regions of the consensus promoter sequences of E. coli were upstream of the presumed initiation codon of the gene. The DNA sequence was most closely related to the aphA-3 gene from Streptococcus faecalis, showing 55.4% sequence similarity. There was 45.6% identity at the amino acid level between the aphA-3 and the aphA-7 proteins. Of the three conserved regions noted previously in phosphotransferase genes, the aphA-7 amino acid sequence was identical to the six conserved amino acids in motif 3, but differed in one of the five conserved amino acids in motif 1 (if gaps are permitted) and 3 of the 10 conserved residues in motif 2. The 32.8% G + C ratio in the open reading frame of the aphA-7 kanamycin resistance gene, which is similar to that of the C. jejuni chromosome, suggests that the aphA-7 may be indigenous to Campylobacters. PMID- 2550984 TI - Cloning and genetic analysis of tra cistrons of the Tra 2/Tra 3 region of plasmid RP1. AB - Transfer-defective mutants of the 10.4-kb Tra 2/Tra 3 region of RP1 were identified by their ability to be complemented by clones carrying all or part of this region. The respective mutations occurred in six cistrons whose order (traA, B, E, R, P, Q) and location were determined by deletion and insertion mapping. The cistrons occupy a minimum of 5.5 kb with the most distal, traA, spanning the 28.0-kb map position and traR the KpnI site at map position 24.1 kb. Each cistron is expressed independently, as Tn5 or Tn504 insertions in any one cistron do not affect the other five. The phenotypes controlled by each cistron suggest that all contribute to pilus biosynthesis/function while three (traB, R, and P) also contribute to surface exclusion. Given the occurrence of tra cistrons in the "silent" region between Tra 2 and Tra 3 we propose that the epithet "Tra 2" should be used to describe this entire region. PMID- 2550985 TI - Sequence and distribution of IS866, a novel T region-associated insertion sequence from Agrobacterium tumefaciens. AB - We have identified a new insertion sequence, IS866, located in the auxin synthesis gene TA iaaH of Tm4, a wide host range biotype III octopine/cucumopine type Agrobacterium tumefaciens strain with two T regions on its tumor-inducing (Ti) plasmid, TA, and TB. IS866 is 2716 bp long, has inverted repeats of 27 bp with three mismatches, and generates 8-bp direct repeats upon integration. In addition to IS866, pTiTm4 carries two copies of a related element, IS867, associated with TA and TB, respectively. A systematic study of 92 virulent Agrobacterium strains has shown that among the three biotypes all octopine/cucumopine and vitopine biotype III isolates contain IS866-like elements. The various octopine/cucumopine Ti plasmids always carry IS866 and IS867 at the same position as in pTiTm4. The chromosomes of the bacteria which contain these Ti plasmids also carry IS866 and IS867 copies but in varying numbers and locations. PMID- 2550986 TI - Specificity of transposition of Tn7 in Vibrio parahaemolyticus. AB - Tn7 was found to transpose at a high frequency from the plasmid, RP4::Tn7, to the chromosome of Vibrio parahaemolyticus. Seven isolates carrying Tn7 insertions were derived from three wild-type strains isolated from geographically distinct areas, and HindIII and BstEII DNA digests of these strains were probed with a ColE1::Tn7 biotinylated probe. The results indicated that V. parahaemolyticus is similar to several other species which have been studied in having a highly preferred site of insertion of Tn7 in the chromosome. PMID- 2550987 TI - The mitochondrial genome of Fusarium oxysporum. AB - Physical and genetic maps have been constructed for mtDNA from strains of the fungus Fusarium oxysporum representing three pathogenically specialized forms. All three mtDNA maps are circular. Their sizes are 45 kb for F. oxysporum f.sp. raphani and 52 kb for both F. oxysporum f.sp. conglutinans and F. oxysporum f.sp. matthioli. The genetic loci for cytochrome b, the mitochondrial 25S ribosomal RNA and cytochrome oxidase subunit II, have been identified and are similarly arranged on the three genomes. PMID- 2550989 TI - The effect of MK-801 and other antagonists of NMDA-type glutamate receptors on brain-stimulation reward. AB - MK-801 is a ligand at phencyclidine recognition sites associated with NMDA coupled cation channels, where it acts as a potent noncompetitive antagonist of central glutamate/aspartate (NMDA-type) receptors. Low doses (10-100 micrograms/kg IP) produced a dose-related and prolonged (greater than 1 h) enhancement of variable-interval self-stimulation responding. Higher doses (300 micrograms/kg) caused flaccid ataxia and disrupted responding. Ketamine HCl (3.0 100 mg/kg IP), a dissociative anaesthetic binding to the phencyclidine site, produced a similar response pattern, but facilitation was less prolonged and occurred over a narrower dose range. Kynurenic acid (3.0-300 mg/kg IP), a nonselective competitive antagonist of glutamate receptors, produced only depression of responding, possibly the result of kynurenate-induced blockade of central kainate and/or quisqualate receptors. The behavioural stimulant effects of MK-801 appear to be an intrinsic and essential feature of selective NMDA antagonists, and these effects of MK-801 differ qualitatively and quantitatively from the well-known facilitatory effects of dopamine-dependent stimulants. PMID- 2550988 TI - Sertraline-induced desensitization of the serotonin 5HT-2 receptor transmembrane signaling system. AB - Sertraline is a new, selective serotonin (5HT) uptake inhibitor with antidepressant activity. The effect of chronic administration of sertraline on 5HT-2 receptors in rat cortex was compared with that of the tricyclic antidepressant, amitriptyline. 5HT-2 receptors were evaluated in binding assays using [3H]-ketanserin and in functional assays of transmembrane signaling, hydrolysis of phosphoinositides. The daily injection of 17 mg/kg sertraline induced a desensitization of 5HT-2-mediated phosphoinositide hydrolysis after 28, but not 21, days. The administration of 1.2 mg/kg/day via continuous release pumps caused a more rapid desensitization. Amitriptyline administered chronically also produced a desensitization of the 5HT-2-mediated phosphoinositide hydrolysis response. A decrease in the density of 5HT-2 binding sites accompanies the functional desensitization after amitriptyline, but changes in 5HT-2 binding sites were not detected after chronic sertraline administration. Studies of the mechanism of action of sertraline show that the desensitization of the phosphoinositide hydrolysis response is homologous in nature, and that it is not secondary to changes in the synthesis of precursor lipids. Other possibilities such as alterations in coupling efficiency or in the activity of effector enzymes are currently being considered. The present results suggest a new postsynaptic action of antidepressant drugs at central 5HT-2 receptors (i.e., changes in 5HT-2 signal transduction at a site distal to the cell surface binding site) and illustrate the importance of studies of receptor signaling pathways to complement radioligand binding. PMID- 2550990 TI - Pituitary--adrenal response to capture in Cayo Santiago--derived group M rhesus monkeys. AB - 113 rhesus monkeys, representing 4 age classes, 3 matrilines, and immigrant adult males in a 161-member Cayo Santiago-derived troop living in a 2-acre enclosure, were sampled for levels of plasma ACTH and cortisol during a period of capture and brief cage confinement for routine veterinary examination. ACTH levels showed significant decreases over initially high values following capture in all subjects except infants, whereas cortisol levels remained elevated throughout the sampling period. Members of the lowest-ranking matriline had significantly higher ACTH levels than members of the other matrilines and immigrant males. Infants and juveniles exhibited higher cortisol levels than adolescent and adult monkeys. The overall pattern of results was generally consistent with previous findings from laboratory studies, providing not only evidence of generality across conditions and subject populations but also the basis for more detailed subsequent analyses of the relationship between pituitary-adrenocortical responsiveness, behavioral response to challenge, and age-sex-dominance status in wild-born rhesus monkeys. PMID- 2550991 TI - [Nuclease-aided simulation of the action of radiation on Chinese hamster fibroblasts. The role of double-stranded DNA breaks induced by restrictases in initiating chromosomal disorders]. AB - Comparison was made between the effectiveness of restrictases inducing double strand DNA breaks with blunt (Hae III and Eco RV) and cohesive (Hind III and Sal I) ends and that of gamma-radiation on the initiation of chromosome aberrations. The analysis of the spectrum of chromosome aberrations induced in the presence or absence of DNA repair inhibitors, as well as the study of the pattern of cell distribution by the number of DNA breaks per cell showed that the decisive role in the initiation of chromosome mutagenesis is played by the localization of the break in certain sequences of target DNA rather than the type of the break. PMID- 2550992 TI - [Postradiation changes of active ion transport systems of the CNS. The effect of superlethal doses of ionizing radiation on the Na,K pump in surviving brain slices]. AB - Active potassium ion transport in surviving sections of rat brain was irreversibly inhibited 6 min, 1 h and 6 h following whole-body single X irradiation. At the same time, the accumulation of products of lipid peroxidation and phospholipase hydrolysis was followed up during the development of radiation pathology. The relationship was noted between the postirradiation changes in the physicochemical status of lipids of the membrane and the impairment of its transport function. PMID- 2550993 TI - [Postradiation changes of the active ion transport systems of the CNS. Na, K ATPase of neurons and neuroglia]. AB - A study was made of the effect of X-radiation (0.31 C/kg and 3.875 C/kg) on Na, K ATPase in fractions enriched with neurons and neuroglia. The results show the impairment of the neuronal-glial relationship in Na, K-ATPase activity. The most important differences in the pattern of changes in Na, K-ATPase system of brain cells were followed up after irradiation with lethal and sublethal doses. PMID- 2550994 TI - [The antibody-producing cell count of rats exposed to polymeric 239Pu]. AB - In experiments with Wistar rats a study was made of the content of antibody forming cells (AFC) in the spleen at remote times (3 to 12 months) after intravenous injection of 239Pu(IV) in doses of 166, 55, and 18 kBq/kg body mass. The doses absorbed in the central and peripheral immunity organs were defined. Pronounced spleen hypoplasia and profound inhibition of humoral immunity were displayed 1 year after the injection of a small amount of the radionuclide. AFC deficiency in animals was amounted to 11-32 per cent throughout the entire period of observation. PMID- 2550995 TI - Adenomatous hyperplastic nodules in the cirrhotic liver: differentiation from hepatocellular carcinoma with MR imaging. AB - Differentiation of hepatocellular carcinomas from adenomatous hyperplastic nodules (AHNs) is important for the early and precise detection of hepatocellular carcinoma in the cirrhotic liver. For this purpose, the authors compared findings on magnetic resonance (MR) images of surgically resected AHNs (n = 7) with those of hepatocellular carcinoma (n = 47). AHNs were divided into two histologic groups: those without atypia (n = 5) and those with atypical hepatocytes or malignant foci (n = 2). All AHNs without atypia were hyperintense on T1-weighted spin-echo images and hypointense on T2-weighted spin-echo images relative to the surrounding liver. However, almost all hepatocellular carcinomas, except for two lesions with massive coagulation necrosis, were demonstrated as hyperintense on T2-weighted images. MR imaging may be useful in the differentiation of AHN without atypia from hepatocellular carcinoma in the cirrhotic liver. PMID- 2550996 TI - Biological effect of bright light. AB - 1. Five minute bright light exposures reduced plasma levels of melatonin in eight normal subjects. 2. No significant change in ACTH levels occurred. 3. These results raise the possibility that short intense light exposures can synchronize circadian rhythms as well as benefit patients with seasonal affective disorder. They also indicate that short pulses of bright light do not affect pituitary ACTH production. PMID- 2550997 TI - M-signal hypothesis--metabolic modulation of membrane functions in rat brain. AB - 1. Monoamine oxidase (amine: oxygen oxidoreductase [deaminating], EC 1.4.3.4) (MAO) and Na+/K+ ATPase (Na+/K+-ATPase, Mg2+-dependent ATP phosphohydrolase, EC 3,1,6,3) showed reciprocal relationship in their activity patterns during experimentally perturbed situations in vivo as well as in vitro. 2. M-signal hypothesis is proposed, which attempts at explaining the regulation of plasma membrane function by the intermediate of metabolic processes in the cell. This metabolic intermediate referred to as M-signal exercise its influence on the plasma membrane Na+/K+ ATPase which is responsible for the uptake process. PMID- 2550998 TI - [Lipiodol UltraFluid in the imaging diagnosis of hepatocarcinoma with cirrhosis]. AB - Fifty patients with HCC associated with hepatic cirrhosis underwent intra arterial injection of Lipiodol UltraFluid (LUF) during diagnostic DSA of liver parenchyma, 42 of them for a complete chemotherapeutic treatment, 8 for an isolated diagnostic control. LUF is known to be specifically captured by HCC neoplastic tissue, with long-term persistence in the lesion if injected in the arterial hepatic tree; this is not the case with other focal hepatic masses. Therefore LUF opacification can be used to demonstrate small daughter tumors not shown by CT or US in cases with evidence of HCC, or to diagnosis HCC in clinically positive patients with no evidence of tumor at non-invasive screening. In our series of patients, accumulation of LUF in the HCC was observed in 100% of the cases, with no false negatives. Two false positives (4%) were observed, due to CT being performed too early (it should be performed not sooner than 10 days after the injection). Overall DSA accuracy was 78%, with 22% false negatives. In 14% of the cases DSA was positive for HCC in patients with aspecific noninvasive screening. CT, performed 10 days after LUF injection, demonstrated HCC daughter tumors not depicted by US, conventional CT, and angiography, in 34% of the cases, and in 9% of the patients only CT/LUF was able to show HCC in clinically positive cases with no evidence of tumor on other imaging techniques. Specificity, sensitivity and over-all accuracy were thus 100% in our series; LUF was well tolerated by the patients, and no technical complications were observed. In our opinion, the diagnostic DSA and CT/LUF is justified only for the typification of suspected focal nodules unsuitable for biopsy: in other instances, especially in case of HCC with positive biopsy/clinical findings and focal nodular mass, the technique should be directly employed as a therapeutic approach, with the injection of lipiodolized agents to treat both primary and daughter nodules after surgery in operable patients, and to begin chemoembolization treatment in patients with intrahepatic polyfocal diffusion. DSA and LUF are therefore of primary importance in the diagnosis and therapeutic flow-chart of HCC associated with hepatic cirrhosis. PMID- 2550999 TI - [Vascular lesions of the ENT organs. Considerations on cases hospitalized at the Coltea ENT Clinic in 1983-1987]. PMID- 2551000 TI - Immunoglobulin gene alterations in human heavy chain diseases. PMID- 2551002 TI - Increased sensitivity to bradykinin in pregnant rats. AB - We tested the effects of late pregnancy on the depressor response to bradykinin (100, 200 and 400ng) in rats. All experiments were performed under anesthesia by intraperitoneal injection of pentobarbital (50 mg/kg). Catheters were connected to the arterial and venous lines for blood pressure recording and administration of drugs. Late pregnancy (19- to 21-day) showed a hypersensitivity of the depressor response to bradykinin. The effects of captopril, Kininase II inhibitor, on the depressor response to bradykinin were examined in rats. Since captopril (50, 100 and 200 micrograms) notably increased the depressor response to bradykinin both in nonpregnant and 19- to 21-day pregnant rats, kininase II acts as a bradykinin metabolizing enzyme in vivo. Captopril (50, 100, 200 micrograms), however, resulted in the augmented parallel increases of depressor response to bradykinin (100ng) in both nonpregnant and pregnant rats. Our data may suggest that kininase II is not involved in the hypersensitivity to bradykinin in late pregnancy in rats. PMID- 2551001 TI - Regional distribution of calcium/calmodulin-dependent phosphatase activity of calcineurin in rat brain. AB - The regional distribution of calcineurin activity (measured using p-nitrophenyl phosphate which detects the phospho-tyrosylphosphatase activity of calcineurin) shows that the striatum, hippocampus and cerebral cortex contains high calcineurin activity. Within the striatum, calcineurin activity does not appear to be present in dopaminergic terminals, since lesions of the nigro-striatal dopaminergic pathway (which reduce striatal dopamine levels by 97%) had no effect on calcineurin activity. On the other hand, kainic acid, which destroys neurons whose perikarya are in the striatum, reduced calcineurin activity by 86% indicating that calcineurin activity is localized in striatal intrinsic neurons. Calcineurin apparently does not exist in glia, since glial cells actually proliferate in kainic acid lesioned striatal tissues. PMID- 2551003 TI - Effect of polyamine related tetraamines on anti-ulcerogenic activity and anti H+,K+-ATPase activity. AB - The relationship between the structure of polyamine related tetraamines and their anti-ulcerogenic activity in rats was studied. Among the tetraamines tested, linear tetraamines had stronger anti-ulcerogenic activity in both restraint stress-induced gastric ulceration and ulceration in pylorus-ligated rats. Macrocyclic tetraamines and polycyclotetraamines had less anti-ulcerogenic activity. Correlation was detected between the inhibitory effect of tetraamines on gastric secretion in pylorus-ligated rats and their inhibitory effect on H+,K+ ATPase activity in rat gastric mucosa. Thermine and spermine were the most effective in these two inhibitions. Thus, it is suggested that primary aminogroups are important in both anti-ulcerogenic activity and anti-H+,K+-ATPase activity. PMID- 2551005 TI - Cardio-pulmonary-cerebral resuscitation. Proceedings of a workshop. Beerse, Belgium, September 19, 1988. PMID- 2551004 TI - A quantitative structure-activity relationship study on some pyrazolo[4,5 c]quinolines acting as inhibitors of benzodiazepine-receptor binding. AB - By a quantitative structure-activity relationship (QSAR) study, the ability of a series of 1-arylpyrazolo[4,5-c]quinolin-4-ones to displace specific [3H] flunitrazepam from bovine brain membranes is shown to be significantly correlated with steric and hydrophobic constants of aryl subtituents, suggesting that substituents of 2- and 6-positions produce dominant steric effects and that those of 3- and 5-positions are involved in strong hydrophobic interaction with the receptor. PMID- 2551006 TI - Pre-CPR conditions and the final outcome of CPR. The Cerebral Resuscitation Study Group. AB - Outcome of cardiac arrest (CA) is very much influenced by pre-CPR conditions. To assess the importance of these pre-CPR factors, an analysis of the Belgian CPCR registry was made according to some pre-CPR conditions. In this registry, several variables related to pre-arrest, arrest, CPR and post CPR period have been recorded in 4548 patients. The pre-CPR conditions studied were: age, witnessed event or not, pre-arrest health state, underlying disease, site of cardiac arrest, type of respiratory arrest and type of cardiac arrest. Age did not influence outcome significantly. The importance of witnessing is very significant. Severe pre-arrest disability reduces chances on long-term survival (LTS) to half and overall health status longterm survivors is clearly less. Intoxication and metabolic origin of CA have good prognosis (LTS, 21%). Trauma/exsanguination, drowning, SIDS and sepsis have bad prognosis (LTS, 1-3%). Cardiac (LTS, 12%) and respiratory (LTS, 14%) origin have similar outcome, although significant difference exists in occurrence of cerebral failure, suggesting that post-ischemic encephalopathy is more severe in respiratory than in cardiac origin. The most frequent site of CA, the home of the patient, has poor outcome results (LTS, 5%). Gasping is significantly related to successful outcome. In the out-of-hospital setting the type of CA was 25% VF (LTS, 77%), 65% asystole (LTS, 4%) and 10% EMD (LTS, 3%). Outcome of the subgroup out-of hospital, witnessed, VF is comparable to other reports. This sub-group seems to us the most appropriate for clinical trials. PMID- 2551007 TI - Tolerance and pharmacokinetics of flunarizine after cardiac arrest. The Cerebral Resuscitation Study Group. AB - Neuronal calcium overloading after complete ischemia-anoxia of the brain might be the primary process initiating chemical cascades which lead to cell death. According to this hypothesis calcium-entry blocking agents act on the final common pathway of brain damage. Flunarizine, a selective calcium-entry blocker (without influence on heart rate and on cardiac contractile force), was administered to 12 unconscious patients, recovering from cardiac arrest (CA) of cardiac origin, according to a strict dose-range infusion protocol. Blood pressure and heart rate (HR) were recorded before, during (t = 10 min, 20 min) and after (t = 30 min, 2 h, 4 h, 6 h, 8 h) each flunarizine infusion (maximum 4 infusions). A significant, although not clinically relevant, decrease in heart rate was noted during the first infusion. Systolic (SBP) and diastolic blood pressure (DBP) also decreased during the infusion without reaching statistical significance. Plasma levels of flunarizine were determined before and after each infusion (t = 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h). Flunarizine plasma concentrations declined very rapidly after cessation of each infusion. Sequential half-lives were in the order of 11-19 min and 5-7 h, and primarily reflect rates of distribution between the systemic circulation and the rapidly equilibrating tissues such as the brain. No substantial accumulation of flunarizine was described and plasma levels were proportional to the give dose. Therefore, flunarizine pharmacokinetics can be considered as linear for doses up to 50 mg. PMID- 2551008 TI - Flunarizine i.v. after cardiac arrest (FLUNA-study): study design and organisational aspects of a double-blind, placebo-controlled randomized study. AB - To investigate the cerebro-protective effect of flunarizine i.v. in patients successfully resuscitated from ventricular fibrillation outside hospital, a double-blind, placebo-controlled randomized study was started in February 1988 in West-Berlin with all eight Mobile Intensive Care Units and ten major hospitals participating. Start of treatment was in the patient's home immediately after restoration of stable circulatory conditions. Primary endpoints reduction of mortality due to hypoxic brain damage and more rapid improvement after cardiac arrest and resuscitation. Inclusion criteria men and women without age limit, ventricular fibrillation prior to or upon arrival of a MICU, primarily successful cardiac resuscitation with continuing unconsciousness. Recruitment of patients will end January 31st, 1990. An inclusion of about 500 patients is expected. PMID- 2551009 TI - Prognostic evaluation of post-cardiac arrest patients in the intensive care. PMID- 2551010 TI - Decision making to cease or to continue cardiopulmonary resuscitation (CPR). The Cerebral Resuscitation Study Group. AB - CPR should be initiated in any patient who has a cardiac arrest. This might improve overall outcome but implies that CPR is started in patients without any virtual chance for long-term survival (LTS). The aim of this study is, by analysing retrospectively 2713 out-of-hospital cardiac arrests (CA), to identify indices which might be of help in the decision making to continue or to discontinue CPR. In an important number of unsuccessful CPR attempts ALS-time did not exceed 20 min. This occurred more frequently in subgroups where limited chances of LTS are expected on clinical grounds. The decision to cease CPR might have been based on other clinical and/or ethical parameters which were not recorded in the registry. This behavior results in a "self-fulfilling prophecy". A subset of patients with limited chances for LTS (0/405) can be identified: patients in electromechanical dissociation (EMD) or asystole on arrival of the mobile intensive care unit (MICU) team, without pupil reaction to light during CPR and with inefficient cardiac massage by the MICU (405/2713). Other patients in EMD or asystole without pupil reaction to light during CPR (1373/2713) but with efficient ECC should be resuscitated for more than 30 min, especially if the patient is gasping during CPR (LTS 27/1373). Patients in EMD or asystole on arrival of the MICU with pupil reaction to light during CPR (236/2713) should have an ALS-time of at least 45 min (LTS 42/236). Cardiac arrests in ventricular fibrillation (VF) (699/2713) should be resuscitated for at least 45 min, especially when gasping during CPR (LTS 119/699). PMID- 2551011 TI - Early prognostic indices after cardiopulmonary resuscitation (CPR). The Cerebral Resuscitation Study Group. AB - An early prediction score (EPS) is constructed as the sum of five events: the type of cardiac arrest is ventricular fibrillation; the type of respiratory arrest is gasping; pupil reaction is unequal, slow or normal, but present; swallowing activity is present and the cardiac arrest has been witnessed. Presence of any of these events contributes one point to the score, while absence contributes nothing to it. EPS during resuscitation results in a comparable amount of information, whether used to predict success, alive and conscious 14 days post-CPR or no-success. EPS early (10 min) after initially successful resuscitation is more effective in predicting no-success than success. EPS during CPR does not allow decision making as far as stopping or continuing CPR efforts. EPS early after CPR does neither allow decision making as far as stopping or continuing critical care efforts after initially successful CPR. EPS does, however, weigh the likelihood of success against that of no-success, which can be used when discussing the chances of the patient with his relatives. PMID- 2551012 TI - United Kingdom resuscitation outcome study. PMID- 2551013 TI - Are inter-center differences in EMS-management and sodium-bicarbonate administration important for the outcome of CPR? The Cerebral Resuscitation Study Group. AB - The hospital of Brugge relies on selection of the emergency calls and sends a Mobile Intensive Care Unit (MICU) whenever cardiac arrest (CA) is suspected. The University Hospital of Leuven does no selection of calls and responds to every emergency call by sending an ambulance with an advanced life support (ALS) trained nurse. The MICU is called when the ambulance crew recognizes the emergency to be a CA. The Leuven system is a so-called tiered system. Although MICU-response times are significantly longer in Leuven than in Brugge, no difference is found as to the success of CPCR. The immediate response to all emergency calls by specialized E.D. nurses (paramedic) capable of ALS, seems to make up for the difference in MICU-response times. The University Hospital of Jette has a higher success-rate for CPCR for in-hospital CA, than the University Hospitals of Leuven. Due to size and lay-out differences, the MICU-response times are shorter in Jette than in Leuven. Basic life support (BLS) provided by doctors and nurses present at the scene, does not seem to be able to compensate for longer MICU-arrival times. The introduction of semi-automatic or automatic defibrillators, to be used by the BLS trained medical and nursing personnel, might be able to make up for the longer MICU-intervention times. Inter-center differences were witnessed as far as the amount of sodium-bicarbonate infused during CPR. Within each group of total duration of CPR an inverse correlation exists between the amount of bicarbonate infused and the success rate of CPCR. Partial correlation between the bicarbonate infused and the survival with regaining of consciousness at 14 days post-CPR, with constant CPR-time, is statistically significant. This indicates that long-term CPCR success is inversely correlated with increasing amounts of sodium-bicarbonate infused. Short duration of CPR and low adrenaline dosage correlate with immediate and long-term success of CPR. On the contrary, low versus high bicarbonate dosage has hardly any influence on immediate success (restoration of spontaneous circulatory activity) but low bicarbonate dosage favours long-term success (survival accompanied by recuperation of brain function). Our data support a negative effect on long-term survival with recuperation of consciousness from infusion of more than 1 mEq/kg body weight of sodium-bicarbonate during CPR. No final conclusions can be drawn so far as to the mechanisms of this negative effect at the level of the brain. PMID- 2551014 TI - Prognostic significance of neurologic examination and glycemia after cardiac arrest. PMID- 2551015 TI - Glycemia in the post-resuscitation period. The Cerebral Resuscitation Study Group. AB - An association between high glycemia on admission after resuscitation from an out of-hospital cardiac arrest and poor neurological recovery has been reported. It remains controversial whether the high glycemia on admission causes the poor outcome or is just an epiphenomenon. The Cerebral Resuscitation Study Group therefore registered the glycemia on admission in 417 patients resuscitated after an out-of-hospital cardiac arrest. Our data confirm that a high glycemia on admission is related to a poor outcome. There is no relationship between the glycemia on admission and the duration of cardiopulmonary resuscitation (CPR). However, there is a positive but weak correlation between the dose of adrenaline administered during CPR and the glycemia on admission. This indicates that the higher glycemia on admission in patients with a poor outcome may, at least in part, be due to CPR parameters, such as the amount of adrenaline used, that are linked with a bad prognosis. However, it cannot be excluded that a high glycemia contributes to the brain damage after cardiac arrest. PMID- 2551016 TI - Serum electrolyte disturbances in the post-resuscitation period. The Cerebral Resuscitation Study Group. AB - In the context of the registration project of the Belgian Cerebral Resuscitation Study Group, the presence or absence of electrolyte disturbances (serum K+ less than 3.0 or greater than 5.5 mEq/l and/or serum Na+ less than 130 or greater than 150 mEq/l) was registered during the 24-h period following resuscitation after an out-of-hospital cardiac arrest. The analysis of 161 consecutive patients seen in the period 1983-1987 at the University Hospital of Gent indicates that patients with such electrolyte disturbances do not have a worse prognosis. Moreover, we also looked at the serum concentrations of potassium and magnesium in 100 and 90 patients respectively by means of a retrospective analysis of the files of 113 consecutive patients seen during 1985-1988 at the University Hospital of Gent and the Free University of Brussels. Hypokalemia (serum K+ less than 3.5 mEq/l) was observed in 30% of the patients and was not related to outcome. The hypokalemia could not be explained by alkalosis; no relationship was found with either the amount of adrenaline administered during resuscitation or the duration of CPR. An abnormal magnesium level (serum less than 1.8 or greater than 2.4 mg/dl) was found in 42% of the patients and our data suggest that the prognosis may be worse in this group. A prospective study on the clinical significance of disturbances in magnesemia would be of interest. PMID- 2551017 TI - Influence of time intervals on outcome of CPR. The Cerebral Resuscitation Study Group. AB - Time is an essential element for successful CPR. Two time factors are particularly important: the duration of complete CA and the time to advanced life support. According to a registration protocol, these time factors, together with other variables and outcome were recorded in 3083 CA cases, treated by the NICU teams of 7 major Belgian hospitals. The mean duration of complete CA is 10.3 min for CPR failure (79%); 5.3 min for initial CPR success (21%); 3.4 min for long term CPCR success (7%). The mean time to ALS is 19.7 min for CPR failure; 14.6 min for initial CPR success: 12.7 min for long-term CPCR success. Both duration of CA and time to ALS are independently related to outcome; the mean duration of BLS is less than 10 min and not significantly related to outcome. Response time of BLS and ALS are increasingly important in their 'rapid response' failure zone (time to ALS more than 8 min, time to BLS more than 4 min). The tiered MICU system, with nurse paramedics before physicians, has equal time to ALS and comparable outcome results to the non-tiered MICU system. According to our present experience, the following time goals are proposed for Belgian EMS-MICU systems: duration of CA less or equal to 4 min (introduction time less than or equal to 1 min and response time of BLS less than or equal to 3 min) and time to ALS less than or equal to 9 min. These time goals stand, in Belgium for perfection of public training in CPR and for spreading of a higher number of MICU teams countrywide. PMID- 2551018 TI - Relevance and significance of pre-CPR conditions in cardio-pulmonary-cerebral resuscitation. A graphic analysis by means of Spectramap. The Cerebral Resuscitation Study Group. AB - The effects of pre-arrest conditions on success rates of CPCR have been studied in 4501 circulatory arrests that have been recorded by the Belgian CPCR-Registry from 1983 to 1987. A graphical technique called Spectramap has been used for the simultaneous perception of clinical relevance and statistical significance. The success of CPCR is determined to a large extent by pre-arrest conditions. Arrests occurring inside hospitals possess a far better prognosis than those observed outside. (Overall success rate inside hospitals is 18% vs. 7% outside). Site of arrest, time to call, time to CPR, and bystander intervention appear to be relevant and significant conditions outside the hospital. Age, underlying disease and degree of disability are shown to be relevant and significant inside hospitals. The Belgian CPCR-Registry, combined with Spectramap, also allows assessment of the efficiency of emergency services and the efficacy of life supporting treatments. This article describes the graphical method of Spectramap and its application to the Belgian CPCR-Registry. PMID- 2551019 TI - Influence of pre-CPR conditions on EMS response times in circulatory arrest. The Cerebral Resuscitation Study Group. AB - Several time intervals, with important influence on the outcome of CA and CPR, are determined by the local EMS-MICU characteristics: time to introduction in the EMS, response time of BLS, duration of BLS before ALS. These time factors have been studied in 2779 out-of-hospital CA cases, treated by the MICU in teams of 7 major Belgian hospitals. The analysis compares the time intervals in the following pre-CPR conditions: the age of the patient; the previous health status of the patient; the disease underlying the CA; the site where CA occurs; the witnessing of the CA; the type of CA; the MICU center, responding to the CA. The mean introduction time is 4.6 min, the mean response time of BLS is 5.1 min, the mean duration of BLS before ALS is 11 min. Introduction in EMS should be improved in CA due to intoxication, drowning, SIDS and respiratory disease, and overall when CA occurs at home. PMID- 2551020 TI - The Belgian Cardiopulmonary Cerebral Resuscitation Registry. Form protocol. The Cerebral Resuscitation Study Group. PMID- 2551021 TI - Bystander cardiopulmonary resuscitation (CPR) in out-of-hospital cardiac arrest. The Cerebral Resuscitation Study Group. AB - Prevalence of bystander CPR and effect on outcome has been evaluated on 3053 out of-hospital cardiac arrest (CA) events. Bystander CPR was performed in 33% of recorded cases (n = 998) by lay people in 406 cases (family members 178, other lay people 228) and by bystanding health care workers in 592 cases (nurses 86, doctors 506). Family members and lay people mainly applied CPR in younger CA victims at public places, roadside or at the working place. Sudden infant death syndrome (SIDS) and drowning are highly represented. Health care workers performed CPR mainly in older patients, at public places or at the roadside and especially in case of cardiac or respiratory origin. CA caused by trauma/exsanguination and intoxication/metabolic origin received less bystander CPR (23% resp. 22%). Cardiac arrests receiving bystander CPR are more frequently witnessed and have a shorter access time to the emergency medical service (EMS) system and shorter response time of basic life support (BLS). Advanced life support (ALS) response time is significantly longer. In witnessed arrests of cardiac origin receiving bystander CPR a significantly better late survival was observed. In non-witnessed arrests of cardiac origin early and late survival are significantly higher in patients receiving bystander CPR. In CA events where response time of ALS exceeds 8 min, the beneficial effect of bystander CPR is most significant. Furthermore no deleterious effect of bad technique or inefficient bystander CPR can be demonstrated. PMID- 2551022 TI - Prehospital CPR: a review, in perspective. PMID- 2551023 TI - Out-of-hospital cardiac arrest: the teaching of experience at the SAMU of Lyon. AB - Because of the improvement resuscitation techniques have shown since the 1960s and because of the development of the out-of-hospital medical care, a cardiac arrest is no longer synonymous with death in every case. However the cardiac arrest resuscitation is only relevant if its adverse consequences can be limited. That is mainly the neurological after-effects and the cellular anoxia. Therefore, the "Service d'Aide Medicale Urgente" (SAMU) of Lyon has been concentrating its research aiming at: (a) Shortening the duration of cardiopulmonary resuscitation to limit the cerebral anoxia. (b) Analysing and treating some of the causes responsible for the aggravation of anoxia. On the basis of several studies in Lyon, here are some suggestions: (1) The use of high doses of epinephrine that unables a better percentage of primary recoveries (47.5% vs. 39%) (P less than 0.05) and secondary recoveries (21.3% vs. 14.8%) (P less than 0.01) without modifying the qualitative survival at long term. (On the basis of: 5 mg intravenous bolus repeated every 3 min in case of asystole instead of 1 mg every 5 min as it is usually recommended). (2) The choice of a peripheral intravenous line instead of a central intravenous line each time it is possible for the administration of drugs since it is as efficient as the second one. (40.7% vs. 33.4%) (P:NS). (3) The alkalinisation of the prolonged cardiac arrest in order to keep the acid-base balance. Most of the survivors show a pH equal or superior to the normal standard. (On the basis of 1 mmol/kg of sodium bicarbonate if the cardiac arrest lasts for more than 10 min). (4) The abolition of the dextrose solution as maintaining infusion the patients who are in a "coma depasse" (brain death) after the resuscitation have an average glycemia superior to the survivors without after-effects. (19.7 vs. 14.8 mmol/l) (P less than 0.05). (5) The monitoring at once at the hospital of the intra-cranial pressure. It reveals the frequency of high pression at an early stage (superior to 15 mmHg in 51.1% of the cases) and the absence of favourable evolution in case of high intracranial pressure. At the moment the absence of consequences on the ICR of a calcium entry blocker (Nimodipine) is being studied. The first results do not seem to show any improvement of the cerebral survival. (6) The prophylactic treatment of septicemia with intestinal origin since they occur frequently and prove to be fatal. PMID- 2551024 TI - Evaluation of cardiopulmonary resuscitation (CPR) techniques. The Cerebral Resuscitation Study Group. AB - The prevalence of different CPR techniques and the use of adjuncts during the resuscitation attempt by the members of the emergency medical service (EMS) system [bystander, emergency medical technician (EMT), ward nurse, tiered nurse or paramedic, mobile intensive care unit (MICU) has been registrated prospectively during a 5-year period by 7 major Belgian EMS systems. A total of 4548 cardiac arrests have been registered, 3083 happened outside and 1465 inside the hospital. Evaluation of the methods used for assessment of quality of the CPR techniques revealed that this approach was biased both by the status of the health care provider and by the outcome of the patient. Nevertheless, it was evident that the well-accepted standards and guidelines for CPR and emergency cardiac care (American Heart Association and Safar) are poorly applied. This seems to be influenced by the qualification, the experience and the training level of the health care provider. In 998 of 3053 studies out-of-hospital arrests (33%) CPR was initiated by bystanders. Of these, 59% were bystanding health care workers. They performed external chest compression (ECC) more frequently than mouth-to-mouth insufflation (MOMO) (poor technique: 16-19%). In 18% the rescuers were family members who applied more MOMO than ECC (poor technique: +/- 50%). Laymen (23%) performed more ECC than MOMO (poor technique: +/- 33%). EMT and ward nurses apply mainly the bag-valve-mask technique. The bag-valve-tube technique is more frequently used by nurses of a tiered system. The MICU-team applies usually the bag-valve-mask prior to intubation. PMID- 2551025 TI - [Surgical treatment of insulinoma. Experience at the Salvador Zubiran National Nutrition Institute]. AB - Eighteen cases of insulinoma treated at the Instituto Nacional de la Nutricion in Mexico City are presented. The cases were operated on between 1959 and 1988, and include 10 women and 8 men with a mean age of 38. The duration of symptoms ranged between 4 and 96 months with a median of 27; 78% of the patients had neuropsychiatric symptoms, 61% seizures, and 55% loss of consciousness. One of the patients had evidence of type I multiple endocrine neoplasia. Diagnosis was established in all the patients with the demonstration of Whipple's triad and also with an elevated insulin determination in the last nine patients. In six cases the insulinoma was enucleated; in two a Whipple procedure was performed; in nine a distal pancreatectomy and in the remaining patient only resection of hepatic metastases; 67% of the tumors were diagnosed as benign solitary adenomas, 16.5% as carcinomas and the same number as hyperplasia. Operative mortality was 5.5%, and morbidity 44%. The three carcinomas survived 6, 8 and 36 months. Disappearance of the hypoglycemic symptoms was obtained in 93% of the cases of adenoma and hyperplasia. PMID- 2551026 TI - [Intracellular messengers in the regulation of renin secretion]. AB - The intracellular messengers that seem to be involved in renin secretion (RS) from juxtaglomerular cells (JG) are calcium (Ca), cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP). Unlike the majority of secretory systems, an increase in intracellular Ca concentration and calmodulin and protein kinase C activation inhibit RS. The intracellular Ca concentration in JG cells can be modified if: 1) the normal mechanisms of Ca extrusion of these cells is altered; 2) the calcium output is blocked by lanthanum; 3) the function of the voltage-sensitive Ca-channels is modified; 4) uptake or liberation of Ca from endoplasmic reticulum is modified; 5) plasmatic membrane is bypassed with calcium ionophores such as A 23187. 6) JG cells are stimulated by hormones that increase Ca and activate protein kinase C such as angiotensin II, vasopressin or alpha-1 adrenergic agonists; 7) extracellular Ca concentration increases or decreases. RS is stimulated by dibutyryl cAMP, cAMP phosphodiesterase inhibitors and by hormones and agents that activate adenylate cyclase (beta adrenergic agonists, bradykinin, histamine, forskolin and ethylcarboxamide adenosine). On the contrary, RS is inhibited by hormones and agents that inhibit adenylate cyclase such as: alpha-2 adrenergic agonists, neuropeptide Y, angiotensin II and cyclohexyladenosine. Pertussis toxin increases basal RS, blocks the inhibition by agents and hormones which inhibit adenylate cyclase and potentiate the stimulation produced by beta-adrenergic agonists. In JG cells, atrial natriuretic peptide inhibits RS, increases cGMP and decreases cAMP. The increase in cGMP correlates well with the inhibition of RS. PMID- 2551027 TI - [Mycoses in otorhinolaryngology. Apropos of 167 cases]. AB - The authors observed 167 cases of E.N.T. mycoses over a 14-year period (1974 1988) in the E.N.T. Departments of the Abidjan University Hospitals in the Ivory Coast, and in two private health institutions in the city. The majority of cases involve candidiasis (91 cases, or 54.5%), followed by aspergillosis (72 cases, 43.1%) and rhinoentomophtorosis (4 cases, 2.4%). Men are more affected than women (125 as against 42). Men suffering from rhinoentomophtorosis are, for the most part, farmers. Among the contributory factors, we found respectively the abuse of antibiotics, either alone or in association with corticoids for general or local use (ear drops), bathing in lagoons, and diabetes. Bacterial infection is often associated with these mycoses--mainly streptococci and staphylococci aurei. Clinical signs are dominated by pruritus, dull pains, a feeling of fullness in the ear, or of burning in the pharynx. An association of systemic Miconazole and Amphotericin B (local use) has given the best results for candidiasis and aspergillosis. For rhinoentomophtorosis, treatment was long, and even disappointing, until the use of Ketoconazole which may without doubt be considered as the medicament of choice. PMID- 2551028 TI - [Laterocervical paraganglioma]. AB - Diagnostic and surgical features of lateral cervical chemodectomas are discussed in relation to a personal series of 16 patients with carotid body tumors and 10 with tumors of the intravagal glomus. Clinically, these affections are poor in symptomatology, those of the carotid glomus presenting as an isolated cervical tumor, intravagal glomus tumors as parapharyngeal masses of the restrostyloid type. Arteriographic findings are discussed in detail, as they constitute the only pathognomonic signs, enable differentiation between the two tumor sites, and supply essential pre-operative data: location of the pedicles, possible invasion of arterial walls, and the limits of the tumor. Surgical approach should be wide to enable control of the distal end of the internal carotid artery, if necessary under the base of the cranium, excision of the tumor usually being possible by employing the sub-adventitial resection technique. PMID- 2551029 TI - [Bordetella pertussis: the principal determinants of its pathogenicity]. PMID- 2551030 TI - The diagnostic value of somatosensory evoked potentials in the diseases of peripheral nervous system. AB - The mean latency value of somatosensory evoked potentials (SEP) recorded at the Erb's point, spinal and cortical level, was assessed in 20 normal subjects by percutaneous stimulation of the peripheral nerves (median, ulnar, superficial radial, tibial and peroneal). At the Erb's point and spinal cervical level, we distinguished the classically described potentials (N9, N13, N14) and the cortical "far-field" potentials such as: P17, N20, P24, N35 and sometimes P45. The central conduction time was calculated by determination of the interpeak latency N9-N13 and N13-N20 (for the upper limb) and LP-P37 for the lower one. A study of 100 patients with peripheral nerve diseases: 30 polyneuropathies, 50 radiculopathies, 9 cases with carpal tunnel syndrome, 5 with brachial plexus injury, 6 with compressive or traumatic diseases of the peripheral nerves, demonstrates the value of the SEPs in the assessment of the nervous lesion site (central or peripheral). In polyneuropathies, a decrease in amplitude and delayed latency of the N9 potential as well as delayed latency of the early cortical potentials on stimulation of the median and tibial nerves occurred. Delayed N9 and low amplitude with delayed latency spinal potential (N13 and LP) were found in radiculopathies. In myelopathies, the central spinal conduction time (N9-N13) was delayed and there were also delayed latencies of the cortical SEPs on lower limb stimulation. The patients with brachial plexus injury had a change in the N9 to N13 amplitude ratio, with prognostic value. Cortical recordings of the SEPs are also of special prognostic value and may suggest the surgical exploration when the axonal functional continuity is lost in brachial plexus injury and compressive or traumatic lesions of the peripheral nerves. PMID- 2551031 TI - Respiratory health of brickworkers in Cape Town, South Africa. Background, aims and dust exposure determinations. AB - The respiratory health and work environment of 575 brickworkers in five brickworks in Cape Town, South Africa, were investigated by means of a questionnaire, a physical examination, pulmonary function testing, chest radiography, and dust measurements. This study presents the background, aims, and methods of the determining dust exposure assignments. The mean concentrations of respirable dust and total dust were 2.22 and 15.16 mg/m3, respectively, with a mean free-silica percentage of 2.1%. Subjective and objective, as well as categorical and continuous, dust indicators were constructed for subsequent analyses, together with findings from the health survey, which are reported elsewhere. PMID- 2551032 TI - Respiratory health of brickworkers in Cape Town, South Africa. Appropriate dust exposure indicators and permissible exposure limits. AB - The predictive value for respiratory abnormalities of dust indicators was assessed for 268 brickworkers. The subjective/objective correlations were poor (r = 0.09). The subjective indicators were found to be predictive of early/mild abnormalities, while the objective indicators were better for more severe abnormalities. The respirable/total dust correlations were high (r = 0.99). Length of service was a good proxy predictor for most respiratory abnormalities, while respirable dust was a good proxy for respirable free silica. Simple screening measures with potential application in less-developed settings, like subjective questionnaire responses and total dust measurement, are suggested. The American Conference of Governmental Industrial Hygienists' threshold limit values for free silica and "nuisance" particulates appear to be too high to prevent the occurrence of abnormalities. The World Health Organization's health-based limit for respirable free silica (1 mg/m3) and hygienic standards of well below 5 mg/m3 would have prevented much of the respiratory abnormality found. Hygienic standards for dust and the concept of "nuisance" dust need to be reevaluated by further research. PMID- 2551033 TI - The cycling of calcium as an intracellular messenger. PMID- 2551034 TI - Cellular aspects of retention and transport of inhaled soluble and insoluble actinide compounds in the rat lung. AB - The binding of 241Am-hydroxide polymers (as models for readily soluble actinide compounds) to the cell components of rat lung was investigated using differential centrifugation, density gradient centrifugation, gel chromatography, carrier-free electrophoresis and electron microscopic autoradiography (with 241Pu). Irregularly shaped and spherical mixed (U, Pu)O2 particles (as models for insoluble actinide compounds) were administered to rats by inhalation and intratracheal installation and the lung and organ retention was determined. Electron microscopic studies were performed with rat lung and with rat and bovine alveolar macrophages exposed to the actinide compounds in vitro. In the case of the mixed (U, Pu)O2 particles the lung retention was independent of the particle shape and route of administration. Whereas 241Am administered as a hydroxide polymer was transferred rapidly from lung to skeleton and liver, only a few percent of the initial alveolar deposit was found in these organs after mixed oxide inhalation. It is concluded that all types of particles are stored primarily within phagolysosomes of alveolar macrophages. In the case of readily soluble compounds, the actinides are solubilized within these lysosomes and become bound to cytosolic ferritin in the alveolar macrophages. They are then released from the macrophages and probably cross the alveolar membranes as transferrin or as low-molecular-weight forms. Insoluble compounds remain within the lysosomes of alveolar macrophages, but there are indications of chemical damage to the lysosomal membranes, causing the particles to lie free in the cytoplasm. PMID- 2551035 TI - Effects of continuous exposure to 239Pu-alpha rays on Co631 cells. AB - Exposure of Co631 cells to 239Pu-citrate leads to continuous irradiation and results in increasing cell death which depends on dose and incubation time of the metal. The D37 is calculated to be 0.13 Gy. Plutonium-239 induces the amplification of the integrated SV40 sequences of the Co631 cells. Other cellular genes such as the alpha-actin gene were not amplified, indicating that the SV40 gene amplification is a selective process. Southern blot analysis of genomic DNA shows that not all S40 sequences are equally amplified. Excised sequences of the SV40 genome only appear after an exposure time of 4 days. PMID- 2551036 TI - Neutrophil Mac-1 and MEL-14 adhesion proteins inversely regulated by chemotactic factors. AB - The neutrophil Mac-1 and gp100MEL-14 adhesion proteins are involved in neutrophil extravasation during inflammation. Both the expression and activity of Mac-1 are greatly increased after neutrophil activation. In contrast, neutrophils shed gp100MEL-14 from the cell surface within 4 minutes after activation with chemotactic factors or phorbol esters, releasing a 96-kilodalton fragment of the antigen into the supernatant. Immunohistology showed that gp100MEL-14 was downregulated on neutrophils that had extravasated into inflamed tissue. The gp100MEL-14 adhesion protein may participate in the binding of unactivated neutrophils to the endothelium; rapid shedding of gp100MEL-14 may prevent extravasation into and damage of normal tissues by activated neutrophils. PMID- 2551037 TI - T cell receptor gene trans-rearrangements: chimeric gamma-delta genes in normal lymphoid tissues. AB - Joining of V-, D-, and J-region gene segments during DNA rearrangements within all antigen receptor genes involves recognition of the same highly conserved heptamernonamer sequences flanking each segment. In order to investigate the possibility that recognition of these conserved sequences may sometimes permit intergenic joining of segments among different antigen receptor genes, DNA of normal human lymphoid tissues was examined by polymerase chain reaction amplification for the presence of chimeric gamma-delta T cell receptor gene rearrangements. These studies detected V gamma-(D delta)-J delta and V delta-(D delta)-J gamma rearrangements in thymus, peripheral blood, and tonsil. Analysis of thymus RNA indicated that many of these rearrangements are expressed as V gamma-(D delta)-J delta-C delta and V delta-(D delta)-J gamma-C gamma transcripts. Most transcripts (19 of 20 complementary DNA clones studied) are appropriately spliced and show correct open translational reading frames across the V-(D)-J junctions. Thus, chimeric antigen receptor genes are generated in a subset of normal lymphoid cells, probably as a result of chromosomal translocations, and such genes may possibly contribute to increased diversity within the antigen receptor repertoire. PMID- 2551038 TI - Long-term potentiation in the motor cortex. AB - Long-term potentiation (LTP) is a model for learning and memory processes. Tetanic stimulation of the sensory cortex produces LTP in motor cortical neurons, whereas tetanization of the ventrolateral nucleus of the thalamus, which also projects to the motor cortex, does not. However, after simultaneous high frequency stimulation of both the sensory cortex and the ventrolateral nucleus of the thalamus, LTP of thalamic input to motor cortical neurons is induced. This associative LTP occurs only in neurons in the superficial layers of the motor cortex that receive monosynaptic input from both the sensory cortex and the ventrolateral nucleus of the thalamus. Associative LTP in the motor cortex may constitute a basis for the retention of motor skills. PMID- 2551039 TI - Type I and type II GABAA-benzodiazepine receptors produced in transfected cells. AB - GABAA (gamma-aminobutyric acid A)-benzodiazepine receptors expressed in mammalian cells and assembled from one of three different alpha subunit variants (alpha 1, alpha 2, or alpha 3) in combination with a beta 1 and a gamma 2 subunit display the pharmacological properties of either type I or type II receptor subtypes. These receptors contain high-affinity binding sites for benzodiazepines. However, CL 218 872, 2-oxoquazepam, and methyl beta-carboline-3-carboxylate (beta-CCM) show a temperature-modulated selectivity for alpha 1 subunit-containing receptors. There were no significant differences in the binding of clonazepam, diazepam, Ro 15-1788, or dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM) to all three recombinant receptors. Receptors containing the alpha 3 subunit show greater GABA potentiation of benzodiazepine binding than receptors containing the alpha 1 or alpha 2 subunit, indicating that there are subtypes within the type II class. Thus, diversity in benzodiazepine pharmacology is generated by heterogeneity of the alpha subunit of the GABAA receptor. PMID- 2551040 TI - Conflict over conflict of interest. PMID- 2551041 TI - NIDA aims to fight drugs with drugs. PMID- 2551042 TI - Tracing hormone action in the cell. PMID- 2551043 TI - Transformation by v-sis occurs by an internal autoactivation mechanism. AB - Transformation by the v-sis oncogene appears to require an interaction of its protein product, p28v-sis, with the receptor for the platelet-derived growth factor (PDGF). However, this interaction may not occur at the cell surface as predicted by the autocrine hypothesis because phenotypic transformation was not reversed by incubation of SSV-NRK cells with antisera to PDGF and because morphological transformation did not occur when nontransformed NRK cells were cultured continuously with p28v-sis. A mutant of the wild-type v-sis gene was constructed that encodes a v-sis protein targeted for retention within the endoplasmic reticulum and Golgi. NRK cells expressing the mutant v-sis gene did not secrete any detectable v-sis protein but were as fully transformed as wild type v-sis transfectants. The results support a mechanism of transformation by v sis in which internal activation of the PDGF receptor occurs before expression of either p28v-sis or the PDGF receptor at the cell surface. PMID- 2551044 TI - [Distribution of substance K in the rat heart and its cardiovascular effects]. AB - The present work was to investigate the distribution of Substance K (SK) in the rat heart and its cardiovascular effects. The content of SK-like immunoreactive material (SKLI), determined by using specific radioimmunoassay, was higher in the atrium (19.9 +/- 3.5 pmol/g tissue) than in the ventricle (4.1 +/- 0.8 pmol/g tissue). SKLI in the heart existed in the nerves fibers and its molecular form was identical with synthetic SK. There were high affinity binding sites of SK on the cardiohybricyte CP 8401: KD = 0.161 nmol/L, Bmax = 8.08 pmol/L. SK stimulated the release of ANF from the cardiohybricytes and from the isolated rat atria. Furthermore, SK injected intravenously induced a hypotensive effect in rats, and decreased left ventricle end systolic pressure (LVESP), +/- LVdp/dtmax, as well as slightly increased heart rate (HR). In isolated Langendorff perfusion heart of rat, SK increased HR, LVdp/dtmax, left ventricle peak systolic pressure (LVPSP) and perfusion pressure (PP). These effects of SK were inhibited by a tachykinin receptor antagonist, (D-Pro2, D-Trp7.9)-SP. These findings suggest that SK exist in the heart and might be able to bind with its specific binding sites on the cardiocytes, thus inducing the release of ANF from the atrium and regulating cardiovascular functions. PMID- 2551045 TI - Case report 525: Benign fibrous histiocytoma (BFH) of thumb. PMID- 2551046 TI - Role of inositol lipid second messengers in regulation of secretion: studies of thyrotropin-releasing hormone action in pituitary cells. PMID- 2551047 TI - Hormonal and neurotransmitter regulation of Ca channel activity in cultured adenohypophyseal cells. PMID- 2551048 TI - [Hormonal changes in chronic diffuse diseases of the liver]. AB - The relationship between adrenocorticotropic hormone as well as renin and potassium activity and blood aldosterone secretion was examined in normal subjects and patients with chronic hepatic diseases. It is demonstrated that aldosterone stimulation is controlled by simultaneous renin-angiotensin (RA) and hypothalamo-adenopituitary effects in normal subjects and patients with chronic active hepatitis (CAH), the RA effects prevailing in normal subjects, and hypothalamo-adenopituitary ones, in CAH patients. Cirrhosis of the liver was associated with the greatest deviations in the stimulant-aldosterone relationship. Viral cirrhosis with ascites featured a considerable RA increase, affecting the adrenals, while the contribution of ACTH was reduced considerably. ACTH level was the highest in patients with alcoholic cirrhosis. PMID- 2551049 TI - [Intrapulmonary administration of metipred in infectious-allergic bronchial asthma]. PMID- 2551050 TI - Paresis of proximal lumbar roots after reduction of L5-S1 spondylolisthesis. PMID- 2551051 TI - Walking and cycling tests in neurogenic and intermittent claudication. AB - The walking and cycling tolerance of 19 patients with neurogenic claudication and 11 with intermittent claudication was assessed, first in the upright and then in 30 degrees of flexion, repeating the tests on a second day. The flexed posture improved the walking and cycling distance, respectively, in 11 and six patients with neurogenic claudication. In two of the 11 patients, this posture significantly improved by more than 100% of both walking and cycling distances. In addition, one patient had claudication pain when he was walking, but could cycle unlimited distance. Only one of the patients with intermittent claudication could walk or cycle further in the flexed position. Although some patients with neurogenic claudication increase their walking distance by flexing forwards, we conclude that posture-related walking and cycling are insufficiently sensitive tests to distinguish between neurogenic and intermittent claudication. PMID- 2551052 TI - Reconstruction of congenital chest-wall deformities. AB - Pectus deformities and Poland's syndrome are two relatively common congenital deformities of the chest wall that are amenable to reconstruction. The extent of the structural deformity in pectus deformity and the degree of associated cardiopulmonary dysfunction are critical variables in preoperative assessment. The operative approaches range from more extensive sternal eversion to the more popular subperichondrial cartilage resection with or without internal fixation. In Poland's syndrome, the options for reconstruction include anterior transfer of the ipsilateral latissimus dorsi muscle through a transaxillary tunnel and attachment to the clavicle and sternum. Submuscular insertion of a mammary prosthesis can be added in the female patient. PMID- 2551053 TI - cAMP increases the rate of GABAA receptor desensitization in chick cortical neurons. AB - During prolonged application of GABA to cultured neurons from the chick embryo cerebrum, whole-cell voltage-clamp recordings show a decline in GABA-gated chloride currents due to desensitization. At a holding potential of -60 mV, desensitization can normally be described as a single exponential process with a time constant (tau) of 7-10 sec at a GABA concentration of 100 microM. After exposure to 50 microM forskolin, the peak amplitude of the GABA-induced currents declined and a fast component of desensitization (tau = 0.92 sec) appeared, whereas the slow component was essentially unchanged. This effect of forskolin was reversible after washing. Similar, although less robust effects were produced by incubation with 8-Br-cAMP, but not by 1,9-dideoxyforskolin. The desensitization process could also be measured by the GABA-dependent uptake of 36Cl- into the cultured neurons. A 20-sec incubation with 10 microM GABA in physiological saline produced a 29% inhibition of subsequent GABA-gated 36Cl- uptake in the presence of 40 mM K+. This inhibition was increased to 64% by the addition of 100 microM forskolin to the preincubation medium. This effect of forskolin was prevented by 100 microM 2',5'-dideoxyadenosine, an inhibitor of adenylate cyclase. A similar acceleration of desensitization was produced by 0.5 mM 8-Br-cAMP or by 0.5 mM isobutylmethylxanthine, but these compounds themselves failed to produce desensitization in the absence of exogenous GABA. In the presence of GABA, cAMP analogs were effective in the order 8-(4-chlorophenylthio) cAMP greater than 8-Br-cAMP greater than N6,O2'-dibutyryl-cAMP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551054 TI - Region-specific loss of alpha 1-adrenergic receptors in rat brain with aging: a quantitative autoradiographic study. AB - The effects of aging on the density and affinity of alpha 1-adrenergic receptors (alpha 1-ARs) were studied in several circumscribed areas of the Fischer 344 male rat brain. Computer-assisted quantitative autoradiography was used to analyze saturation binding isotherms of [125I]BE-2254, a selective alpha 1-AR antagonist. Significant decreases in receptor density of 15 and 29% were observed in the thalamus at 16-18 and 24-28 months of age, respectively, when compared to 3-4 month-old controls. Progressive declines in receptor density of 24 and 44% were also found in the olfactory tubercle. In the cerebral cortex, a significant 26% loss in receptors occurred only in the oldest age group. No changes were found in any of the other brain areas investigated, including the cerebellum, brainstem, caudate-putamen, and several subregional areas of the hippocampal formation. Kd values ranged from 12 +/- 1.8 pM in the brainstem to 23 +/- 1.6 pM in the thalamus and were not affected by aging in any area examined. It is concluded that the density of alpha 1-ARs in the Fischer 344 rat brain is diminished with aging in a region-specific manner and that loss of these receptors may account for age-related functional deficits only in a few brain areas. PMID- 2551055 TI - GR 43175: a preferential 5-HT1D agent in monkey and human 1rains as shown by autoradiography. PMID- 2551056 TI - [The Na+/H+ antiport: a target for new antihypertensive agents]. AB - The sodium-hydrogen (Na+/H+) antiport is electroneutral and amiloride-sensitive. It exchanges Na+ for H+ with a coupling ratio of 1:1 and therefore, it has important cellular functions: intracellular pH and cell volume regulation, control of the cell differentiation and growth. Its activity is increased in hypertension. The responsibility of the Na+/H+ antiport in essential hypertension may result in increased vascular tone and vascular hyperplasia. Amiloride is a specific inhibitor of this exchanger but under physiological conditions, very high concentrations of this drug (that are unlikely to be attained in vivo) are required to inhibit the activity of the Na+/H+ antiport. Certain of its analogues are much more potent in vitro than amiloride. Therefore, the Na+/H+ antiport may become a target for new antihypertensive drugs. PMID- 2551057 TI - Plasmin's peptide-binding specificity: characterization of ligand sites in alpha 2-antiplasmin. AB - The basis for specific binding of plasmin to alpha 2-antiplasmin (AP) was analyzed by preparing overlapping synthetic peptides of 11, 17, 18, 19, 26, 33, and 40 amino acid residues corresponding to the carboxy-terminal sequence of AP. Affinities of the peptides for plasmin were estimated by competitive inhibition of the association of AP with plasmin. Dissociation constants with increasing peptide length were: 200, 54, 19, 18, 9.8, 4.7, and 2.8 microM, respectively. Peptides blocked binding sites on plasmin, not the catalytic site, as evidenced by lack of effect on the hydrolysis of chromogenic substrates. Substituting arginine for lysine at the carboxy-terminus or the 17th residue from the carboxy terminus decreased the affinity of peptides for plasmin 9-fold and 5-fold, respectively, implicating these lysine residues of AP as major ligand sites for plasmin. Several stepwise increases in affinity of peptides for plasmin as peptide length increased up to 40 residues suggest contributions by additional sites, possibly other lysine residues. A potential plasmin binding site in fibrin, analogous to that in AP, is identified by affinity for plasmin of synthetic peptides corresponding to part of the alpha-chain ending with residue 207. To explain these data, we propose that plasmin recognizes physiological ligands by binding two or more lysine residues which are optimally presented to favor simultaneous interaction with separate lysine-binding site in plasmin. PMID- 2551058 TI - The effect of low dose of 12-O-tetradecanoyl-phorbol-13-acetate on collagen platelet interactions. AB - We and others have reported that phorbol ester doses ranging from 0.25 to 25 micrograms/ml induce human platelets to aggregate and release serotonin. In this paper, we report the effect of low doses of phorbol (0.5 to 50 ng/ml) on subthreshold amounts of collagen induced platelet aggregation. The platelet aggregation induced by the addition of subthreshold amounts of collagen can be enhanced by low doses of phorbol. The combined low doses of phorbol and subthreshold amounts of collagen induced platelet aggregation can be inhibited by the addition of aspirin and imidazole. The increase in platelet aggregation induced by the combined low doses of phorbol and subthreshold amounts of collagen is probably mediated by the collagen fibril formation. PMID- 2551059 TI - Transient aggregation resistance of human blood platelets in fresh plasma. II. Physiological relevance: influence of fish oil. AB - In order to evaluate the physiological relevance of the blood platelet "transient aggregation resistance" (TAR) test we studied the effect on this test of two different amounts of fish oil, corresponding to .75 g g (2.5 mmol) and 1.5 g (5 mmol) of eicosapentaenoic acid respectively, added in a cross-over design to the normal diet of 16 healthy male volunteers. It appeared that the "baseline aggregation resistance" (BAR), equivalent to the classic platelet aggregation ADP threshold, was not influenced by Maxepa while, in contrast, a significant prolongation of TAR occurred. Apparently platelet aggregation analysed early after blood withdrawal, measures aspects of physiological relevance which, due to their short half life, are missed in the original method. PMID- 2551060 TI - Antithrombotic effect of thiopurinol. AB - Antithrombotic activity of ten pyrazolo pyrimidine derivatives was tested in mouse pulmonary thromboembolism model. Out of these compounds, Thiopurinol (C5H4N4S) showed dose-dependent protection in mice from death/paralysis induced by collagen + adrenaline. It also caused dose-dependent inhibition of thrombus formation in the cat. Thiopurinol inhibited aggregation of platelets induced by ADP and arachidonic acid but did not inhibit superoxide generation. It had no antiinflammatory activity nor any effect on cardiovascular system. The results indicate that the antithrombotic activity of the compound is mediated via inhibition of platelet aggregation. PMID- 2551061 TI - Studies on the human plasma kallikrein-kinin system: alpha-kallikrein does not directly activate blood neutrophils. AB - alpha-Kallikrein was prepared using an improved purification protocol (Burger, D., Schleuning, W.-D. and Schapira, M. (1986) J. Biol. Chem. 261, 324-327) and was employed to reevaluate our previous observations indicating that kallikrein activates blood neutrophils by a mechanism requiring an uncleaved Mr 52,000 NH2 terminal heavy chain. Cellular activation was evaluated by measuring neutrophil aggregation, release of both vitamin B12 binding capacity and myeloperoxidase, and generation of superoxide anion. Whereas all these indicators were evoked by exposing neutrophils to f-Met-Leu-Phe, phorbol myristate acetate, zymosan activated serum, or the calcium ionophore A 23187, we show here that neutrophils incubated with alpha-kallikrein remained unactivated. Moreover, we demonstrate that this lack of activation is accompanied by an inability of neutrophils to specifically bind alpha-kallikrein. PMID- 2551062 TI - Group A streptococci bind human plasmin but not other structurally related proteins. AB - Certain group A streptococci are known to possess a receptor for the human enzyme plasmin. Plasmin is a member of a super gene family that includes other serine proteases and kringle containing proteins. In this study we have examined the interaction of a group A streptococcus with structurally related proteins, including plasmin, glu-plasminogen, tissue plasminogen activator, kallikrein, factor XII, prothrombin, thrombin, trypsin, and urokinase. Our studies indicate that only the key fibrinolytic enzyme, plasmin, demonstrates significant binding activity to the group A streptococcus. PMID- 2551063 TI - Partially N-desulfated heparin as a non-anticoagulant heparin: some physico chemical and biological properties. AB - A partially N-desulfated preparation of heparin (UFH) obtained by thermally inactivating heparinic acid for 24 hours at 50 degrees C (TIHA) was examined for its physico-chemical and biological properties in vitro and in vivo. TIHA has a molecular weight of 14,700 and 27% remaining N-sulfate groups (UFH = 17,500; 100% N-sulfate groups). TIHA has no anticoagulant activity measurable by conventional amidolytic or clotting tests. However, in a rabbit stasis-induced thrombosis model and using two different thrombogenic stimuli (Feiba and PCC(Konyne)/RVV), TIHA afforded a dose-dependent (1.0-2.5 mg/kg) protection sufficient to impair thrombosis (UFH: fully effective at 0.13 mg/kg). TIHA did not produce any bleeding at supramaximal antithrombotic dosage in a rat tail bleeding and a rabbit ear blood loss model and it did not augment ADP-induced aggregation of platelets. In contrast, a completely N-desulfated derivative of UFH (Inoue and Nagasawa, Carbohydr. Res. 46, 87-95, 1976) also lacking measurable in vitro activity was completely inactive in vivo. The results in this study suggest that TIHA may be considered as a non-anticoagulant heparin still retaining antithrombotic activity and also with lower haemorrhagic effect than UFH. PMID- 2551065 TI - The Na+/H+ exchange is required for platelet aggregation induced by AVP (arginine vasopressin). PMID- 2551064 TI - Purification and characterization of plasma protein C inhibitor. AB - Plasma protein C inhibitor (PCI) was purified to homogeneity (greater than 95%) with good recovery (greater than 25%) and reproducibility, and the inhibition of a number of blood clotting and fibrinolytic enzymes by purified PCI was studied. PCI inhibited activated protein C (APC), two-chain urokinase (2c-uPA), two-chain tissue plasminogen activator (2c-tPA), thrombin, factor Xa, plasma kallikrein and factor XIa, and this inhibition was accelerated by heparin. The inhibition of each enzyme was accompanied by formation of enzyme inhibitor complexes and by degradation of the inhibitor to lower molecular weight derivatives. Plasma kallikrein and factor XIa cleaved PCI of native Mr = 57,000 into two products with Mr = 54,000 and 52,000 whereas the other enzymes converted the PCI to a product with Mr = 54,000. PCI did not detectably inhibit alpha-factor XIIa or plasmin. Kinetic studies using PCI yielded the following second-order rate constants for inhibition of human APC, 2c-uPA, 2c-tPA, thrombin, factor Xa, kallikrein and factor XIa respectively: 0.65 x 10(4), 0.22 x 10(4), 0.08 x 10(4), 0.61 x 10(4), 2.01 x 10(4), 6.50 x 10(4), and 9.03 x 10(4) M-1s-1 in the absence of heparin and 1.58 x 10(6), 0.43 x 10(6), 0.03 x 10(6), 0.52 x 10(6), 0.09 x 10(6), 0.18 x 10(6) and 0.74 x 10(6) M-1s-1 in the presence of optimal concentrations of heparin. The rate constants for the inhibition of factor XIa and 2c-uPA by PCI suggest a possible role of PCI in the physiologic regulation of these enzymes. The second order rate constants for inhibition of bovine APC and Gla-domainless bovine APC by human PCI were 0.61 x 10(4) and 0.26 x 10(4) M-1s-1 in the absence of heparin and 0.54 x 10(6) and 0.71 x 10(6) M-1s-1 in the presence of heparin, respectively. Calcium ions (0.05 to 4 mM) did not affect these rate constants. The results obtained with normal and Gla-domainless APC indicate that the Gla domain of APC is not required for inactivation by PGI and is not essential for the heparin stimulation of this reaction. PMID- 2551066 TI - Determination of functional and antigenic protein C inhibitor and its complexes with activated protein C in plasma by ELISA's. AB - Enzyme-linked immunosorbent assays (ELISA's) were developed for the measurement of protein C inhibitor (PCI) antigen and activity and for its complexes with activated protein C (APC) in plasma. For PCI activity and antigen, APC or anti PCI, respectively, was immobilized to microtiter plates and PCI bound was detected with labelled anti-PCI antibodies. For APC:PCI complexes, two different antibodies directed against protein C and PCI were used. The assays for PCI were calibrated with pooled normal human plasma (NHP) and with purified PCI, and for APC:PCI complexes with known concentrations of purified pre-formed complexes added to buffer or to plasma. The lower limit of sensitivity of the PCI activity and antigen assays was 10 ng/ml and 0.5 ng/ml, respectively and for plasma APC:PCI complexes 12 ng/ml. Mean coefficients of variation of 1.5% to 5.8% (intra assay) and 2.1% to 9.8% (inter-assay) were found for the assays. For PCI antigen, a range of 56% to 162% of the NHP value was obtained in samples from 70 healthy donors (mean +/- SD = 98.6% +/- 23.1%). For PCI activity, the range was 59% to 148% (94.3% +/- 20.2). A good correlation (0.92) was obtained when both assays were compared. Plasma levels of APC:PCI complexes in 30 normals were under the detection limit (less than 12 ng/ml). In plasma samples from 10 patients with disseminated intravascular coagulation (DIC) PCI antigen concentrations were decreased (55.6% +/- 20%) and 8 of the patients had APC-PCI complex levels between 32 and 240 ng/ml (median, 35 ng/ml). After addition of 20 micrograms/ml APC to NHP or to protein C depleted plasma, 6.1 micrograms/ml complexes were recovered after 90 min incubation. Incubation of 10 micrograms/ml APC with NHP in the presence of 10 U/ml heparin yielded 11 micrograms/ml complexes after 90 min, which represent more than 90% of the maximum possible value. Thus, the method should be adequate to study complexes of APC in vivo in clinical conditions in which activation of protein C pathway may occur. PMID- 2551067 TI - Cytoplasmic pH dependency of Na+/H+ exchange in human platelets activated with thrombin, arachidonic acid, A23187 and TPA. AB - The cytoplasmic pH (pHc) of human platelets was lowered to 6.8-7.2 by treatment with various doses of nigericin (K+/H+ ionophore), then these platelets were stimulated with thrombin, arachidonic acid (AA), A23187 or 12-o tetradecanoylphorbol-13-acetate (TPA), to monitor the pHc changes using a pH sensitive fluorescent dye, BCECF. The pHc increased with the stimulation in an amiloride-sensitive manner only when the resting-pHc was lower than a certain value (pHc 6.99 for thrombin-stimulation, 6.95 for AA, 7.04 for A23187 and 6.95 for TPA, n = 3). At a higher resting-pHc, the pHc decreased. Similar observations were found using platelets acid-loaded by Na-acetate treatment. These results suggest that stimulation-induced activation of the Na+/H+ exchanger depends on cytoplasmic H+ concentration, and it can function properly only when the resting pHc is lower than about pHc 7. PMID- 2551068 TI - Inhibition by tranexamic acid of the conversion of single-chain tissue plasminogen activator to its two chain form by plasmin: the presence on tissue plasminogen activator of a site to bind with lysine binding sites of plasmin. AB - The addition of tranexamic acid inhibited the conversion of single chain tissue plasminogen activator (sct-PA) to its two chain form (tct-PA) by plasmin. Increase in the concentrations of tranexamic acid resulted in more inhibition of the conversion, with 50% inhibition being obtained at 0.251 mM of the concentration of tranexamic acid. The addition of the fragments of plasminogen such as kringle 1 to 3 (K1-3), and K4 resulted in the facilitation of the conversion of sct-PA to tct-PA by plasmin. The addition of tranexamic acid to the mixture of sct-PA and K4 inhibited the rate of the conversion of sct-PA by plasmin. These results suggest that binding of lysine binding sites of plasmin or plasminogen with sct-PA enhanced its conversion to tct-PA by plasmin, and that there is a site on a t-PA molecule to bind to plasminogen or plasmin. PMID- 2551069 TI - Characterization of GTP-gamma-S binding to isolated human platelet plasma membranes and its relationship with the stimulation of a phospholipase C activity. AB - Binding parameters for the interaction of GTP-gamma-[35S] with isolated platelet plasma membranes have been studied. Analysis of the data by a non-linear curve fitting program indicates that the interaction can be satisfactory described by a model with a single, high affinity binding site (Kd = 0.3 +/- 0.07 microM and Bm = 0.4 +/- 0.2 nmoles of GTP-gamma-S/mg of membrane protein). Binding is selectively inhibited by GDP-beta-S and GMP-PNP (1 microM), but not affected by ATP, CTP, ITP, or UTP, even at mM concentration. Optimal conditions for the interaction were 30 degrees C and pH 8.0. Incubation of the isolated membranes with GTP-gamma-S results in a measurable phospholipase C activity (as detected both by a breakdown of phosphoinositides and an increase of inositide phosphates) which under our experimental conditions is only slightly enhanced by addition of cytosolic proteins. Our results indicate that platelet plasma membranes contain all the necessary elements for signal transduction through the diacylglycerol/inositolphosphates pathway. PMID- 2551070 TI - Subcutaneous treatment of deep venous thrombosis with low molecular weight heparin. A dose finding study with LMWH-Novo. AB - Treatment of deep venous thrombosis with low molecular weight heparin (LMWH-Novo, Logiparin) was carried out with two different doses of Logiparin, 75 XaI U/kg b.w. twice daily and 150 XaI U/kg b.w. once daily subcutaneously for 5 days. Simultaneously warfarin was given from the first day of heparin treatment. Mean age of the twenty patients was 65 years and one third was females. No serious side effects, hematomas, pulmonary emboli or signs of recurrent thrombosis occurred during treatment with either dose regime. Venografic assessment with Marder scoring one week after initiation of Logiparin treatment showed a slight not significant improvement apparent in 40% of the patients. The activities of F IIaI and F-XaI in the blood plasma were found to increase after injection of Logiparin. These two parameters seem to be the most suitable for monitoring the effect during treatment. For future studies on the therapeutic effect of Logiparin in deep venous thrombosis a single dose of 150 to 200 F-XaI activity per 24 hours seems to be most suitable. PMID- 2551071 TI - Methylphenidate and pemoline do not cause depletion of rat brain monoamine markers similar to that observed with methamphetamine. AB - Methylphenidate (Ritalin) and pemoline (Cylert) are central nervous system stimulants which are widely prescribed for attention deficit and other psychiatric disorders. Several other related stimulants, including amphetamine and methamphetamine, have been shown to cause long lasting decreases in monoamine markers in rat brain, characteristic of axonal degeneration. To assess the neurotoxic potential of methylphenidate and pemoline, we compared the effects of multiple injections (sc, bid for up to 4 days) of methylphenidate (21 and 50 mg/kg) and pemoline (20 and 70 mg/kg) with methamphetamine (5 and 15 mg/kg) on rat brain norepinephrine, dopamine, and serotonin levels and transport sites. While decreases were observed in all brain monoamine markers measured in rats treated with methamphetamine, no changes were observed in animals treated with methylphenidate as compared to saline-treated controls. Pemoline failed to induce significant changes in the level of monoamine transport sites; however, a wide array of changes were observed in the levels of 5-hydroxyindoleacetic acid, dopamine, and norepinephrine in various brain areas after a 3-day treatment regimen with a high dose (70 mg/kg) of pemoline. The lack of changes in monoamine transport sites following the repeated administration of high doses of methylphenidate and pemoline suggests that these drugs do not affect axonal integrity. However, the pattern of changes observed in the levels of monoamines after pemoline treatment may have relevance to the self-injurious behavior seen in these animals. PMID- 2551072 TI - Uptake and distribution of 14C during and following inhalation exposure to radioactive toluene diisocyanate. AB - Inhalation of toluene diisocyanate (TDI) results in toxic responses ranging from pulmonary irritation to immunological sensitization. The use of radioactively labeled isocyanate has made it possible to follow the initial uptake of the compound into the bloodstream independent of the final fate of the isocyanate. This study shows that the rate of uptake into the blood is linear during exposure to concentrations ranging from 0.00005 to 0.146 ppm and that the uptake continues to increase slightly postexposure. It also demonstrates that the radioactivity clears from the bloodstream to a level corresponding to approximately a 100 nM concentration of tolyl group after 72 hr and persists at a nanomolar level even 2 weeks following the exposure. This is similar to the response previously reported by this group for radioactively labeled methyl isocyanate. The initial rate of 14C uptake is also a linear function of the concentration of TDI when expressed either as concentration (ppm) or as concentration multiplied by duration of exposure (ppm.hr). This is discussed in comparison with the toxic responses as a function of both ppm and ppm.hr. Finally, the inclusion of the data on methyl isocyanate indicates that the uptake into arterial blood is a function of exposure concentration, independent of isocyanate structure. PMID- 2551074 TI - Nephrotoxicity assessment by measuring cellular ATP content. I. Substrate specificities in the maintenance of ATP content in isolated rat nephron segments. AB - To clarify the characteristics of cellular ATP synthesis in individual nephron segments for assessing nephrotoxicity of chemicals, cellular ATP content was measured by the luciferin/luciferase system under various conditions using intact nephron segments isolated from male Sprague-Dawley rats. Increasing the duration of collagenase treatment of kidney slices significantly lowered the cellular levels of ATP newly synthesized from 2 mM glutamine in PST at 37 degrees C over 30 min (p less than 0.01). The tubular incubation time significantly affected the cellular ATP content in the early and middle portions (S2) of the proximal tubule (p less than 0.05 and p less than 0.01, respectively) over 20 min and in the late proximal tubule over 10 min. Among numerous substrates tested, such as D-glucose, glutamine, pyruvate, DL-lactate, and beta-hydroxybutyrate, the substrate utilization for maintaining cellular ATP content was entirely variable according to each nephron segment. Pyruvate and glutamine were the best substrates in the proximal tubule. On the other hand, ATP production from glutamine was less than that from the other substrates in the distally located nephron segments: medullary and cortical thick ascending limbs of Henle's loop (MAL and CAL, respectively), distal tubule, cortical and medullary collecting tubules (CCT and MCT, respectively). In general, glucose, pyruvate, and lactate appear to be equivalent in maintaining ATP content in the distal segments of renal tubules. A monovalent cation ionophore, monensin, at 10 micrograms/ml decreased the cellular ATP content in MAL, CAL, and MCT significantly. Mercuric chloride (HgCl2) was used as a model compound to study nephrotoxicity by investigating its effects on cellular ATP metabolism in microdissected nephron segments. HgCl2 at 1 x 10(-6) M significantly decreased ATP content only in S2 (p less than 0.05), clearly demonstrating S2 to be the most sensitive segment within the nephron. These results indicate that measurement of cellular ATP content would be a useful method forecasting the intrarenal toxic site and potency of possible nephrotoxic chemical compounds. PMID- 2551073 TI - The effect of organotin compounds on chloride secretion by the in vitro perfused rectal gland of Squalus acanthias. AB - The effects of various organotins on membrane function and electrolyte transport were studied in the marine elasmobranch, Squalus acanthias. The isolated perfused rectal gland was used as a model of electrolyte transport. This gland can be stimulated to secrete chloride by atrial natriuretic peptide, veratrine, and vasoactive intestinal polypeptide although the mechanism of action of each secretagogue is different. By analysis of the inhibitory effect of an organotin in the presence of each secretagogue, the mechanism of inhibition can be inferred. Tributyltin (TBT) produced a reversible inhibition of epithelial transport at 10(-8) to 10(-7) M which resulted from inhibition of stimulus secretion coupling in VIP-containing neurons within the gland. The transporting epithelial cells were unaffected at these concentrations. Trimethytin (TMT) produced inhibition at 10(-7) M which was not reversible and which affected primarily the transporting epithelial cells. Triethyltin and triphenyltin were without effect. The inhibitory effect of TBT and TMT was not affected by simultaneous administration of dithiothreitol. TBT also produced inhibition of oxygen consumption, Na+,K-ATPase, and proton ATPase in dispersed rectal gland cells. These results indicate that organotins are toxic to cell membrane functions which are intimately involved in the movement of electrolytes. This is the first evidence of toxicity to membrane transport functions in a marine species which is at risk from environmental exposure. PMID- 2551075 TI - Altered methylation of ras oncogenes in benzidine-induced B6C3F1 mouse liver tumors. AB - The B6C3F1 mouse is a hybrid strain which exhibits a high (30%) spontaneous hepatoma incidence and sensitivity to chemical induction of liver tumors. The spontaneous hepatoma incidence of the paternal C3H/He strain is approximately 60%, while that of the maternal C57BL/6 strain is very low. The presence of activated oncogenes, primarily Ha-ras, and to a lesser extent, Ki-ras, has been reported in B6C3F1 mouse liver tumors. Because alterations in a gene's capacity for expression, as well as mutation, may be involved in oncogene activation, this investigation was directed toward an examination of a putative control point for transcription, i.e., the methylation state of a gene. Hypomethylation is believed to be necessary, but not sufficient, for transcription. It was therefore hypothesized that alterations in the methylation state of the Ha-ras and Ki-ras oncogenes may facilitate the aberrant expression of these genes in B6C3F1 mouse liver. Restriction enzyme analysis (MspI, HpaII, and HhaI) was used to assess the extent of DNA methylation. MspI digestion of B6C3F1 and C3H/He DNA revealed the absence of a 15-kb Ha-ras band present in MspI-digested C57BL/6 DNA, suggesting that the Ha-ras oncogene of B6C3F1 and C3H/He mouse liver lacks a methylated site. In other respects, the Ha-ras and Ki-ras oncogenes are methylated to a degree which suggests that these oncogenes have a low potential for expression in normal mouse liver. The methylation state of the Ha-ras and Ki-ras oncogenes was also assessed in benzidine-induced hepatomas and adjacent nontumor tissue from B6C3F1 mice. In four out of four cases, the Ha-ras oncogene was hypomethylated in tumor as compared to nontumor tissue and increased expression of the gene was detected in three out of four hepatomas; the Ki-ras oncogene was hypomethylated in two out of four cases. These results suggest that hypomethylation of oncogenes may provide an epigenetic mechanism for facilitating their aberrant expression. The lack of a methylated site observed in the Ha-ras oncogene in B6C3F1 and C3H/He mouse liver may indicate an increased potential for its expression which could, in part, account for the high propensity for hepatoma development in these two strains. PMID- 2551076 TI - Pentachlorobutadienyl-L-cysteine uncouples oxidative phosphorylation by dissipating the proton gradient. AB - A very early event in the toxicity of pentachlorobutadienyl-L-cysteine (PCBC) to rabbit renal proximal tubules is uncoupling of oxidative phosphorylation (R.G. Schnellmann, E. A. Lock, and L. J. Mandel (1986), Toxicologist 6, 176; (1987), Toxicol. Appl. Pharmacol. 90, 521). The mechanism of PCBC uncoupling of mitochondrial oxidative phosphorylation has been investigated using isolated rabbit renal cortical mitochondria (RCM). PCBC increased state 4 respiration of RCM respiring on pyruvate/malate or succinate in a concentration (10-100 microM)- and time (1-5 min)-dependent manner. PCBC also increased state 4 respiration in the presence of oligomycin, an inhibitor of F0F1-ATPase. The effect of PCBC on mitochondrial proton permeability was determined by measuring passive mitochondrial swelling. After a 2-min exposure to PCBC, RCM swelled when placed in NH4Cl or NaCl, but not KCl or sucrose. The protonophore carbonyl cyanide p trifluoromethoxyphenyl hydrazone (FCCP) (1 microM) produced similar effects. After 5 min, RCM swelled when placed in NH4Cl, NaCl, or KCl, but not in sucrose. Aminooxyacetic acid, an inhibitor of cysteine conjugate beta-lyase, blocked the effects of PCBC on respiration, indicating that PCBC can be metabolized by RCM to produce RCM toxicity. These results show that PCBC initially uncouples oxidative phosphorylation by dissipating the proton gradient. Subsequently, additional ion permeabilities occur. These results are in complete agreement with previous observations in rabbit renal proximal tubule suspensions. PMID- 2551077 TI - Inhibition of rat brain Na+-K+-ATPase by triterpene glycosides from holothurians. AB - The effect of triterpene glycosides from holothurians on Na+-K+-ATPase of rat brain was investigated. The marine glycosides are irreversible inhibitors of the enzyme with an average I50 value of 10(-4) M. ATP had a low protective effect against inhibition. The inhibitory effect was increased by preincubation with MgCl2. There was alteration of the activation curve of Na+-K+-ATPase by NaCl and KCl in the presence of glycosides. Triterpene glycosides inhibited the K+ phosphatase activity, but to a smaller degree than the ATPase activity. Na+-K+ ATPase of pig kidney was less sensitive to the marine triterpene glycosides than the brain enzyme. The marine glycosides did not alter the specific binding of [3H]-ouabain to the Na+-K+-ATPase. PMID- 2551078 TI - Physicochemical characteristics of interaction of toxic triterpene glycosides from holothurians with rat brain Na+-K+-ATPase. AB - High-angle X-ray diffraction spectra showed that triterpene glycosides form crystalline complexes with membrane cholesterol. Electron microscopy demonstrated a decreased vesicle size, of the membrane preparation from rat brain which is enriched in Na+-K+-ATPase, by the triterpene glycosides. The Arrhenius plot was linear in the presence of triterpene glycosides. The half-width of the phosphatidylcholine N-methyl proton line in proton NMR spectra was not altered in the presence of marine glycosides. The excimer formation of pyrene, a hydrophobic fluorescent probe, was significantly decreased by triterpene glycosides. The increase of tryptophanyl residue fluorescence demonstrated a change of the Na+-K+ ATPase conformation after treatment with cytotoxic glycosides. PMID- 2551079 TI - Radioimmunoassay of delta-toxin from B. thuringiensis: correlation with bioassay. AB - A radioimmunoassay procedure, developed for delta-toxin of B. thuringiensis, is applicable to the assay of the toxin's bioactivity. When delta-toxin is inactivated to different degrees by heat treatment, the amount of toxin as determined by radioimmunoassay is proportionally decreased. It thus appears that the delta-toxin antibody may recognize and bind to the conformation in the molecule associated with its insecticidal activity. PMID- 2551080 TI - Direct induction of MHC class I, but not class II, expression on endothelial cells by cytomegalovirus infection. AB - MHC expression on endothelial cells after cytomegalovirus infection was investigated using double antibody radioimmunoassays and immunofluorescence flow cytometry. After CMV infection there was an increase in class I and beta 2 microglobulin expression. This increase corresponds to the percentage of cells that are infected with CMV. There was no evidence that the increased class I expression was due to soluble factors produced by the infected cells. Although CMV only affects approximately 10% of the endothelial cells, class I and beta 2 microglobulin expression on the cell population exceeds that induced by an optimal dose of alpha-interferon. Class II expression is not increased after CMV infection. However, when gamma-interferon was added to the medium after CMV infection a normal increase in class II expression was found, suggesting that CMV infection does not inhibit the gamma-interferon-induced expression of these molecules. PMID- 2551081 TI - A phytophile responds... PMID- 2551082 TI - How were introns inserted into nuclear genes? AB - There is now abundant evidence that many introns have been inserted into nuclear genes after the divergence of multigene families, sometimes in a semi-regular pattern with respect to pre-existing domains. This note examines ways in which these insertions might have occurred using known molecular mechanisms. PMID- 2551083 TI - [Cooperation concerning admission to and discharge of elderly people from the hospital. 1. The coordinated contributions of home care personnel]. AB - In cooperation between the home care system in the Municipality of Rodovre and four hospital departments in the County of Copenhagen, a controlled and randomized investigation was undertaken among the hospitalized patients of 65 years and over from the Municipality of Rodovre. A nurse employed by the Municipality particularly for this project visited the hospital daily and followed the 135 participants in the intervention group with the objects of obtaining information from the primary sector if necessary, discussing discharge with the patient and the hospital staff, coordinating possible supportive measures in the patient's home and visiting the patient in his home immediately after discharge from hospital in order to ensure continuity in care and treatment. The 138 participants in the control group underwent the usual procedures in connection with admission to hospital. In this article, the consequences of the contributions by the health visitor in the hospital are described in relation to the course of hospitalization. The intervention group had an average stay in hospital of 11.0 days as compared with 14.3 days in the control group (p greater than 0.05), and the total number of bed-days were 1,490 and 1,970, respectively. In addition, the two groups were compared as regards the number of diagnostic procedures during hospitalization, the number of deaths, the diagnoses on discharge and the functional capacity. No differences were observed in these respects between the two groups. Three patients from the control group were discharged to residential institutions as compared with none in the intervention group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551084 TI - [Cooperation concerning admission to and discharge of elderly people from the hospital. 2. The course 1 year after discharge]. AB - In connection with the health visitor's work in a hospital, a controlled and randomized investigation was undertaken among hospitalized patients of 65 years and over from the municipality of Rodovre. The efforts made in hospital were described in a previous article. Out of the intervention group of 135 patients, 125 were discharged alive. Of these, 114 were visited on one or more occasions immediately after discharge with the object of discussing the process of admission and review of the health and social circumstances to ensure adequate and relevant supportive measures. A total of 125 persons from a comparable control group were discharged alive after having received the usual offers of assistance during and after hospitalization. The two groups were followed for one year after discharge as regards the number of deaths and the utilization of hospital, waiting departments and the municipal residential institutions and home help and home nursing services. No differences were found between the groups as regards the number of deaths, number of re-admissions and the duration of hospitalization. A total of three persons from the intervention group and 14 from the control group utilized wasting departments, nursing homes or residential places in day and night homes. Altogether these persons utilized 774 and 2,136 days, respectively, in institutions during the year after discharge. The intervention group employed more home-help hours and visits from home visitors. This attempt at co-ordination thus appears to the capable of reducing the needs for municipal residential places in patients with considerable loss of function. PMID- 2551086 TI - [Brachial irradiation plexopathy]. AB - Irradiation neuropathy is a term for the damage to peripheral nerve tissue due to irradiation. Brachial irradiation plexopathy is irradiation neuropathy affecting the brachial plexus. This is most frequently a complication of irradiation therapy for cancer of the breast. The incidence varies considerably and is lowest with low total doses of irradiation and limited fractions. The latent period varies from months to several years. The neurological manifestations are paraesthesiae in the fingers, pain, hypaesthesia, hypalgesia, disaesthesia, paresis, hyporeflexia, muscular atrophy and possibly vegetative disturbances. Horner's syndrome may occur. Lymphoedema is observed in approximately on third of the patients. The course of brachial irradiation plexopathy is progressive. No specific treatment is available. The diagnosis is based on the case history, clinical picture, electrodiagnosis and CT of the brachial plexus region. The most important differential diagnosis is metastatic infiltration in the brachial plexus. These two conditions are differentiated best by means of CT guided surgical exploration and histological examination of the tissue. The irreversible nature of brachial irradiation plexopathy and its marked resistance to treatment are such that the optimal irradiation hygienic rules must be observed. PMID- 2551085 TI - [Persons aged 65 years and over admitted to the medical department of a hospital in the Danish provinces with 4 specialized departments. An analysis of extra days of hospitalization related to the services for senior citizens in 4 communities]. AB - In a retrospective investigation of year 1985, it was found that out of 438 medical patients greater than or equal to 65 years there were in all 42 patients who together were responsible for 1,543 bed-days after the time when they were considered to have concluded hospital treatment. The extra bed-days correspond to approximately 9% of the total medical bed capacity. The patients came from four communities whose coverage with nursing homes, home nursing services and home help arrangements were very different. It was found that increase in nursing home capacity was followed by decrease in the number of extra bed-days among the patients who had extra bed-days. When the number of home nurses increased, fewer elderly patients from the community concerned, were admitted who ended by having extra long hospitalization. These conditions are discussed and the results are compared with previously published investigations. It is concluded that improved cooperation between communities and hospitals, is required. PMID- 2551087 TI - [Cooperation concerning admission to and discharge of elderly people from the hospital. 3. Functional capacity, self-assessed health and quality of life]. AB - As part of a total project concerning description and effects of the coordinating efforts of home nurses in connection with admission to and discharge from hospital of persons over the age of 65 years, the functional capacity and the subjective parameters: quality of life, self-assessed health and loneliness were registered on discharge and two months later. In order to assess the effect of intervention, the intervention group was compared with a control group. No differences in the functional capacity could be demonstrated on discharge or after two months. No differences could be demonstrated in the subjective parameters either. The effect of the home nurses' efforts in the total project could be measured as a reduction in the number of institution-days in the intervention group. These results suggest that, in further attempts to reduce the risk of complications in recently discharged elderly patients, further assistance of the functional capacity should be added to the forms of intervention described. PMID- 2551088 TI - [Dihydroergotamine and orthostatic hypotension in elective hip surgery]. AB - The authors undertook a prospective double-blind, controlled trial to investigate the effect of prophylactic low-dose heparin/dihydroergotamine, low-molecular heparin/dihydroergotamine and a placebo on the blood pressure and postural hypotension on postoperative mobilization. A material of 120 patients admitted for hip surgery was subdivided at random into three groups. Examination for postural hypotension was undertaken on the fifth postoperative day. No significant differences were present between the three groups as regards age distribution, systolic and diastolic blood pressures and pulse rates. This investigation did not reveal any significant effect of the antithrombotic combined treatment with heparin/dihydroergotamine on the blood pressure or postural hypotension on postoperative mobilization following elective hip surgery. No cases of ischaemic side effects were observed. However, this investigation cannot disprove a slight hypotensive effect and, for this reason, this treatment should be contraindicated in patients with circulatory disturbances. PMID- 2551090 TI - [Adverse effects in children treated with ACTH in infantile spasm]. AB - During a period of ten years, 13 children were treated for infantile spasms with ACTH. Eleven of these developed Cushing-like appearance, 9 irritability, 7 hypokalaemia, 6 metabolic alkalosis, 5 hypertension, 5 infections and 5 osteoporosis. This investigation reveals that osteoporosis is a more frequent side effect than previously recognized and, similarly, that hypertension is also a common side effect. Routine x-ray control of the thoracic spine, regular measurement of blood pressure and control of urinary calcium and serum calcium, phosphate, sodium and potassium are therefore recommended. As cases of ureteronephrolithiasis have also been observed, examination of the urine for blood and ultrasound examination of the kidneys for formation of calculi should also be considered. PMID- 2551089 TI - [Plasmapheresis in the treatment of peripheral HIV neuropathy]. AB - Peripheral neuropathy following HIV infection is frequently seen among HIV positive-, ARC- and AIDS patients who often complain of debilitating paresthesias. Indirect immunofluorescence technique showed antibodies directed either to myelin and/or axons and to endothelial cells of the blood vessels within the nerve tissue. Sural nerve biopsies from five patients showed demyelination, axonal degeneration and perivascular cellular infiltration. Two of the patients had nuclear inclusions and high anti-CMV titer. Plasma exchange was undertaken on five HIV infected patients which resulted in complete disappearance of symptoms in three, incomplete remission in one and no effect in one patient. The finding of autoantibodies against different nerve tissue components, cellular infiltration, positive effect of the plasma exchange and relapse suggests that autoimmunity may play a pathogenetic role in some of the HIV infected patients with peripheral neuropathy. PMID- 2551091 TI - Prevalence of the plasma bovine leukaemia virus blocking factor in cattle from a commercial dairy herd. AB - The prevalence of the plasma bovine leukaemia virus blocking factor was examined in a commercial herd. The detection of the blocking factor activity depended on the nature of the infected target cells used in the assay. Using the sensitive bovine leukaemia virus-infected target lymphocytes, the factor was found to be significantly more prevalent in infected cattle than in virus-free cattle. Furthermore, in infected cattle, no correlation was observed between the level of bovine leukaemia virus antibody and the presence of the blocking factor. PMID- 2551092 TI - Serological response to a booster foot-and-mouth disease vaccination with strains different from the primary vaccine. AB - Young calves were vaccinated with belgian foot-and-mouth disease (FMD) vaccine and revaccinated with either the same vaccine or with a foreign FMD vaccine. There was a significant serological response to the primary vaccine strains after the first vaccination which was greater following revaccination. At one and two months after revaccination there was no significant difference between the responses to revaccination with vaccine identical to the primary vaccine or with the foreign FMD vaccine. It was concluded that revaccination of young calves is effective even with an FMD vaccine different from the primary vaccine. PMID- 2551093 TI - Viral haemorrhagic disease in rabbits: a review. AB - An acute haemorrhagic disease of rabbits was first reported in a southern province of China in 1984. It subsequently spread rapidly to South China and some parts of North China. The disease is characterized by an acute onset with fever, rapid respiration and sudden death. There is a high morbidity and mortality rate. The pathological changes are consistent with severe generalized circulatory dysfunction (hyperaemia, congestion and haemorrhage), marked degeneration of parenchymatous tissue, pronounced serous-haemorrhagic pneumonia and extensive disruption of reticulo-lymphoid tissue. The disease has been named rabbit viral haemorrhagic disease and it has been suggested that the aetiological agent is a picornavirus. A tissue-derived vaccine has been prepared by homogenizing the liver, lung, spleen and kidney of infected rabbits and inactivating with formaldehyde. This review summarizes the information on the aetiology, and epidemiology and clinical and pathological aspects of this new rabbit disease. PMID- 2551094 TI - Fluorescence anisotropy of UV-irradiated viruses. AB - The steady-state fluorescence anisotropy measurements on influenza and parainfluenza viruses, showed no changes in the microviscosity of the viral membranes after exposure to UV-irradiation, when a fluorescent probe was used, but the intrinsic fluorescence of viral proteins presented, under the same experimental conditions, a significant difference of anisotropy behaviour in the two viruses used. PMID- 2551095 TI - Marek's disease (MD). XX. Incidence and epizootology of MD in broiler flocks. AB - The frequency of Marek's disease in broiler chickens as measured by gross evidence of tumors at processing differs markedly among flocks and between poultry farm origin. Organs affected in broilers, in order of frequency of tumors, were proventriculus (57.6%), spleen (42.3%), kidney (30.6%), lung (19.1%), liver (15.3%), gonads (12.3%), heart (7.6%). Virus isolations were performed on samples from three broiler flocks to examine the possibility that such differences in MD incidence were causally related to differences in the frequency of naturally occurring, nononcogenic serotype 2MD virus infection. Serotype 2 virus was isolated from two flocks. MD condemnations were lower in two serotype-2 infected flocks (V1) than in the third one serotype 1 and serotype 2 positive flock (Mh). Thus, these data suggest a strong positive association between MD losses, housing and a weak probable negative association between serotype 2 virus and MD losses in broilers. However, low level of leukosis condemnations in certain flocks from the same farm are not related to high serotype 2 virus exposure. PMID- 2551096 TI - Epstein-Barr virus (EBV). X. Incidence of EBV antibodies in patients with malignant tumors. AB - Serum samples from 31 patients with various types of malignancies, 18 patients with different viral infections and 6 healthy subjects as controls, were tested by indirect immunofluorescence (IF) method for antibodies against viral capsid antigens (VCA) and the presence of active EBV infection. EBV antibodies anti-VCA were detected in 19 patients with tumors, in 8 patients with viral infections and in 2 healthy subjects. EBV active infection was found out in 9/19, 3/8 and 0/2 EBV anti-VCA positive patients with malignancies, different viral infections and healthy subjects respectively. PMID- 2551097 TI - Further evidence for the presence of A antigen on group B erythrocytes through the use of specific exoglycosidases. AB - Certain antibodies have shown the ability to detect small amounts of A antigenic structures on certain group B cells. These rare cells that reverse group as B, are designated here as B(A) cells. Among the anti-A antibodies capable of detecting these cells are MHO4 (an IgM murine monoclonal antibody) and polyclonal anti-A derived from blood group O donors. The latter (anti-A, B) have been adsorbed exhaustively with normal B cells, to deplete the serum of antibodies to group B antigen. The cell specificity detected by these antibodies can be removed only by an alpha-N-acetylgalactosaminidase (A-zyme) but not by an alpha galactosidase (B-zyme). Inhibition studies show that these reactions can be inhibited by A secretor saliva and cannot be inhibited by B secretor saliva. Moreover, papain treatment of normal group B cells not previously agglutinable with these antibodies, now causes these cells to become reactive, and this specificity, too, is removed only by A-zyme. These results suggest that low levels of blood group A antigen are being recognized by these antibodies and that these structures can exist not only on B(A) cells but on all group B erythrocytes. PMID- 2551098 TI - [The regional contractility of the left ventricle in patients with ischemic heart disease and concomitant hypertension]. AB - Differential evaluation was carried out of hypertrophy as a factor possibly of compensatory or that of furthering disturbances of the coronary circulation and contractile function of the myocardium. Echocardiography, radionuclide ventriculography tests with physical loads revealed that only moderate hypertrophy of the left ventricle in IHD patients with hypertensive disease (grade II) may be considered as one of the compensatory factors maintaining the functional state of the left ventricle myocardium. PMID- 2551099 TI - [2 cases of histiocytoma of the lung]. PMID- 2551100 TI - [Regulation in the expression of alpha-galactosidase gene in raf operon in Escherichia coli]. AB - The alpha-galactosidase, coded for by the first structural gene rafA in the plasmid determined raf operon was an inducible enzyme. In contrast to lac or mel operon, raf operon has more strict structural specificity for inducers. The enzyme can be induced by melibiose and raffinose, or weakly by D-galactose, but not by structurally related sugars such as lactose, PNPG etc.. The alpha galactosidase forming capacity as function of growth curve reached a single peak at the end of the logarithmic phase of the growth. The structure and regulation of raf operon is similar to those of lac operon. The repressormor-mediated negative control plays a major role in the regulation of raf operon, and cAMP-CAP mediated positive control is also involved in the regulation. When 0.4% glucose was added into the medium with other carbon sources, the expression of the enzyme was repressed by 2-3 fold. Transient catabolite repression has been observed neither in inducible nor constitutive alpha-galactosidase expression. Based on alpha-galactosidase assay, in mutant strains CA8306(cya) and CA8445 (cya, crp) the expression level of raf operon was only 9% and 2.5% of that in wild type strain respectively. The glucose effect or the repression in cya mutant can be abolished by 1-5 mmol cAMP. The constitutive alpha-galactosidase expression in cya and cry double mutant (CA8445) remains repressible by glucose, but irreversible by cAMP, suggesting cAMP-CAP complex is not the exclusive mediator of the catablite repression.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551101 TI - Prophylactic activity against Sendai virus infection and macrophage activation with lipophilic derivatives of N-acetylglucosaminylmuramyl tri- or tetrapeptides. AB - The efficacy of N-acetylglucosaminyl-beta (1----4)-N-acetylmuramyl tri- or tetrapeptides (GM) and the lipophilic derivatives for host augmentation against Sendai virus infection and for macrophage activation in vitro was examined. The anti-infectious activities of GM derivatives were shown to increase with the chain length of the fatty acid combined with the diaminopimelyl group. When the macrophages were activated with 1 U ml-1 murine interferon gamma (IFN-gamma) and 0.001 microgram ml-1 GM derivatives, the cytocidal ability of macrophages depended on the length of the side chain, and exhibited a positive relationship with the anti-infectious activity of GM derivatives against Sendai virus infection. These results indicated that the increment of lipophilicity of GM derivatives would play an important role in the anti-infectious activity and macrophage activation in vitro. PMID- 2551102 TI - Efficacy of an inactivated oil-adjuvanted rotavirus vaccine in the control of calf diarrhoea in beef herds in Argentina. AB - We have assessed the potency of an inactivated oil-adjuvanted rotavirus vaccine in beef herds in Argentina. Two different vaccine trials were conducted. In a small-scale experimental trial, involving 21 pregnant cows (13 vaccinated and eight unvaccinated controls), a significant increase in neutralizing antibody titres against different serotypes of bovine rotaviruses was found in both the colostrum and serum of vaccinated cows compared with that of unvaccinated controls. Seven days after birth, half of the calves born to vaccinated dams or to control cows were challenged with live virulent virus whereas the other half of both groups were left in contact with the infected calves in order to mimic a natural field challenge. Although no statistically significant differences in the rate of protection were observed among the different groups of animals, a larger number of vaccinated calves were protected in comparison with their controls, particularly where animals in contact with infected calves were concerned. Secondly, a large-scale field trial was carried out in 17 beef herds involving a total of 4066 vaccinated pregnant cows. In 11 farms morbidity and mortality in calves from vaccinated cows were compared with historical data from the previous 3 years at the same locations. In the other six herds, control groups were used to compare data of the same year: 1540 cows were vaccinated and 2700 were left as controls. Taking into account the previous and current incidence of diarrhoea, morbidity and mortality were significantly reduced in 16 of the 17 beef herds tested. Vaccine effectiveness was also evident in farms where other enteropathogens such as cryptosporidium and coronaviruses were present, together with rotavirus. PMID- 2551103 TI - Messenger RNA stability in yeast. PMID- 2551104 TI - Functional exploration of the yeast (Saccharomyces cerevisiae) genome: use of a mini-Mu transposon to analyse randomly cloned DNA sequences. AB - The development of mega-sequencing techniques requires new methods for global functional analysis of cloned DNA fragments. We have developed a mini-Mu transposon adapted to yeast cloned DNA fragment analysis. This transposon allows us to do the following in a single construction: (i) to probe yeast cells for the presence of expressed open reading frames (ORFs) in the cloned DNA fragment; (ii) to localize these ORFs in the fragment and determine their transcription orientation; (iii) to use beta-galactosidase protein fusions to study regulation of these ORFs; and (iv) to disrupt the corresponding chromosomal genes. On a 5-kb yeast DNA sequence, we have verified the reliability of this new tool by comparing the data obtained with the mini-Mu transposon to those obtained by classical methods. This transposon should be of immediate use in the yeast genome sequencing programme. PMID- 2551105 TI - Modulation of the migration and chemotaxis of PMN cells by hyaluronic acid. AB - Hyaluronic acid (HA) has a dose-related inhibiting effect on the migration and chemotaxis of polymorphonuclear leucocytes (PMN) in vitro. These effects were measured with a new indirect quantitative assay. On average 1 mg HA/ml causes an inhibition to about 80% of the control (spontaneous migration). This effect increased progressively with an increasing HA concentration, and with 4 mg HA/ml only about 19% of the PMN were able to migrate in the in vitro system. Similar results were obtained in the presence of a potent chemotactic factor (leukotriene B4 [LTB4]). In the mean 1 ng LTB4/ml alone stimulated the chemotaxis of PMN by a factor of 3 compared to the spontaneous migration. The highest HA concentration (4 mg/ml) reduced the number of migrating PMN cells to about 17%. From these experiments it may be concluded that the HA in the synovial fluid of the healthy joint has a protective effect against the invasion of PMN cells. This functions is disturbed in inflamed joints by the decrease in the HA concentration and possibly by its depolymerization. The intraarticular application of high molecular weight HA might be an important therapeutic regimen to restore the natural barrier against PMN migration, also in the presence of chemotactic factors and could therefore be helpful for interrupting the inflammatory cascade. PMID- 2551106 TI - [Use of direct calorimetry for continuously following the course of experimental infections in the mouse]. AB - To obtain information about early symptoms of infection measurements of heat production were performed in experimentally infected animals. As a model Mengo virus-infected mice were used. Construction and physical properties of the flow calorimeter device are described. Fever was not detected by method used. However, deviations from normal behaviour of the animals were demonstrated as a result of the relationship between motoric activity and heat produced. Furthermore, the method may be particularly useful for the determination of the exact time of death, which is of interest for mathematical modelling of survival data (RECKNAGEL et al. 1986; GUTHKE et al. 1987; SUHNEL und VECKENSTEDT 1989). PMID- 2551108 TI - Predicting ovulation in the mare on the basis of follicular growth and serum oestrone sulphate and progesterone levels. AB - Follicular growth and hormone determinations were used to predict ovulation in the mare. Thirty Finnhorse mares were used for the investigation and 38 oestrous cycles were studied. The mares were examined by rectal palpation and ultrasonography every 6 hours during late oestrus. Daily blood samples were obtained for progesterone and oestrone sulphate determination. The preovulatory follicle grew 3 mm a day up to 2 days before ovulation. The size then remained constant, before diminishing by 2-3 mm during the last 12 hours. The maximal diameter of the follicle was 43 +/- 4 mm. In 89% of the follicles the round shape became more irregular before ovulation. During the last 24 hours before ovulation 37 of 38 follicles were regarded as mature on rectal palpation. The oestrone sulphate level was highest 24-48 hours before ovulation, the first decrease being observed most commonly around ovulation (+/- 1 day). The size of the follicle was the most reliable criterion in the prediction of ovulation. PMID- 2551107 TI - [Possible involvement of the renin-angiotensin system in reproduction. III. Occurrence and role in pregnancy]. AB - To clarify a possible participation of the renin-angiotensin-system (RAS) in physiological processes of pregnancy, findings are listed about occurrence and variations of RAS components in uterus, placenta, amniotic fluid, fetal membranes, fetus and serum. There exist complete and independent RAS in the respective organs and tissues. During pregnancy concentrations and/or activities of the components change, but in different ways and non-uniforming. The information now available does not yet allow a definite view on the role of RAS during pregnancy and on the correlations to estrogens, progesterone, chorionic gonadotropin, prostaglandins etc. Supposedly RAS may be involved to a high degree in the local regulation of blood flow and the maintenance of fetal vascular homeostasis. Furthermore, there are relations between fetoplacental components of RAS and pregnancy-induced hypertension. Further investigations are necessary. They are expected to offer therapeutic possibilities for influencing pathological processes in which RAS plays a role. PMID- 2551109 TI - The activities of phosphatases and aminotransferases in the epithelium of the small intestine and caecum of white rock cockerels during starvation. AB - Alkaline phosphatase (ALP, EC 3.1.3.1), acid phosphatase (ACP, EC 3.1.3.2), aspartate aminotransferase (ASAT, EC 2.6.1.1) and alanine aminotransferase (ALAT, EC 2.6.1.2) were measured in the mucosal homogenates of the duodenum, jejunum and caecum of full-fed (control), starved and refed White Rock Cockerels. Starvation caused a significant (p less than or equal to 0.05) increase in the activity of ACP in all three segments of the intestine. Subsequent re-feeding brought the activity back to the control level. In contrast ALP activity fell in the duodenum during starvation and was partially restored by refeeding. In the jejunum and caecum the ALP activity decreased during starvation and was fully restored by re feeding only in the caecum. ASAT activity increased (p less than or equal to 0.05) during the entire period of starvation in all three segments. Re-feeding failed to decrease the enzyme activity within 48 hours. Starvation caused a reduction (p less than or equal to 0.05) in the activity of ALAT and re-feeding did not increase the activity in the duodenum and jejunum. The caecum showed no change in the activity during fasting. PMID- 2551110 TI - Malignant luteoma in a sow. AB - A case of malignant luteoma was found in a 3-year-old Landrace sow. A large neoplastic mass was present in the left ovary and small neoplastic nodules were seen in the right ovary. There were metastatic lesions in the diaphragmatic peritoneum and heart. The neoplastic cells were extremely pleomorphic and among them relatively large cells predominated. The neoplastic cells had lipid droplets, membranous structures, intermediate filaments and lipofuscin granules in their cytoplasm. PMID- 2551111 TI - [A modification of the ABC method (avidin-biotin-peroxidase complex) for the detection of viral antigens in coronavirus infections in cats (FIP) and parvovirus type 2 infections in dogs]. AB - A modified ABC-method (avidin-biotin-peroxidase complex) is established to demonstrate viral antigens in paraffin sections of tissues of coronavirus (FIP) infected cats and parvovirus type 2 infected dogs using monoclonal antibodies. The pros and cons and potential sources of error are described and discussed. PMID- 2551112 TI - [The territorial and age-related characteristics of the distribution of patients with non-A, non-B viral hepatitis infected via the fecal-oral mechanism in foci of elevated viral hepatitis morbidity in the Uzbek SSR]. AB - The study of patients from 10 foci of acute viral hepatitides for the presence of HBsAg (in the passive reverse hemagglutination test) and anti-hepatitis A virus IgM (in the radioimmunoassay) has shown high frequency and variability in the spread of hepatitis non-A, non-B, the prevalence of adults aged 20-29 years and children aged 2-4 years among persons involved into the epidemic process and the tendency towards an increase in the proportion of hepatitis non-A, non-B in the total number of cases of viral hepatitides in the republic. PMID- 2551113 TI - [The epidemiological problems of viral non-A, non-B hepatitis with a fecal-oral mechanism of infection transmission]. AB - The results of the study of the epidemiological regularities of viral hepatitis non-A, non-B with the fecal-oral mechanism of the transfer of infection in one of the regions of the Turkmenian SSR are presented. These results have been confirmed by serological and virological laboratory methods. The territories, groups of the population (age groups) and the time of the maximum epidemic risk of infection have been determined. The main (primary) role of the water route (polluted ground water) in the spread of this infection has been proved. PMID- 2551114 TI - The contribution of 99mTc-pyrophosphate tomoscintigraphy in the evaluation of necrotic myocardial lesions. AB - Diagnostic contribution of Conventional Planar Scintigraphy (CPSc) and Single Photon Emission Computed Tomoscintigraphy (SPECT), using 99mTc-Pyrophosphate (99mTc-PYP), have been compared in 49 patients with either transmural (T.I., N = 40), or non-transmural (N.T.I., N = 9) infarction. Sensitivity ratios were, respectively for CPSc and SPECT, 75% and 100% in T.I., 78% and 88% in N.T.I. CPSc was non-diagnostic in 6 out of 49 patients, SPECT in only 1. Extent of the necrotizing process was better delineated on SPECT studies than on CPSc. Specificity of 99mTc-PYP tomoscintigraphy evaluated in a group of 6 young (under 40 years old) normal patients was 6/6 (100%). In nine out of ten additional patients without proven infarction who had underwent contrast coronarography within two weeks after a positive 99mTc-PYP SPECT study, significant stenosis (greater than 50%) were found on arteries supplying blood to pathologic territories as demonstrated on tomoscans. It is concluded that 99mTc-PYP SPECT is a very sensitive and specific way--better than CPSc--in the diagnosis as well as in the evaluation of the extent of necrotic myocardial process. PMID- 2551115 TI - Fine needle aspiration cytologic findings in a case of carotid body paraganglioma (chemodectoma) PMID- 2551116 TI - Cervical koilocytotic atypia and human papillomavirus infection. PMID- 2551117 TI - Sensitivity of urethral cytologic diagnosis of human papillomavirus infection in men. PMID- 2551118 TI - The development of understanding of the mechanism of action of antidepressant drugs. PMID- 2551119 TI - CGS 8216 antagonizes effects of benzodiazepines on defensive-escape and other behaviour during intraspecies conflict in mice. PMID- 2551120 TI - Crystalloid inclusions in folliculostellate cells of adenohypophysis following estrogen withdrawal from primed male rats. AB - Crystalloid inclusions were observed within nongranulated, folliculostellate pituitary cells 72 after estrogen was withdrawn from primed male rats. Like crystalloid inclusions in macrophages reported by others, those observed in this study were associated with structures that appeared to be lysosomes. We suggest that crystalloid inclusions may be the result of phagocytosis, a function of folliculostellate cells proposed by us in a previous study. PMID- 2551121 TI - Metabolism of vitamin D2 and vitamin D3 in patients on anticonvulsant therapy. AB - We examined the effect of short-term treatment with pharmacological doses of vitamin D2 or vitamin D3 on the serum concentration of 1,25(OH)2D metabolites in epileptic patients on chronic anticonvulsant drug therapy. Nine patients were studied before and after treatment with vitamin D2 4000 IU daily for 24 weeks and 10 before and after treatment with vitamin D3 in the same dose. Before treatment the serum concentrations of 1,25(OH)2D and 25(OH)D were significantly lower in epileptics than in normal subjects (P less than 0.01). Vitamin D2 treatment increased the serum concentration of 1,25(OH)2D2, but a corresponding decrease in 1,25(OH)2D3 resulted in an unchanged serum concentration of total 1,25(OH)2D. The serum concentration of 25(OH)D2 and 25(OH)D increased significantly, whereas there was a small decrease in 25(OH)D3. Vitamin D3 treatment did not change the serum concentration of 1,25(OH)2D3 whereas serum 25(OH)D3 increased significantly. The correlation between the serum ratio of 1,25(OH)2D2/1,25(OH)2D3 and 25(OH)D2/25(OH)D3 estimated on vitamin D2-treated epileptic patients and normal subjects was highly significant (P less than 0.01). The data indicate that the serum concentration of 1,25(OH)2D2 and 1,25(OH)2D3 are directly proportional to the amount of their precursors 25(OH)D2 and 25(OH)D3 and that the concentration of total 1,25(OH)2D is tightly regulated. PMID- 2551122 TI - Spectrum of morphological appearance of amyloid deposits in Alzheimer's disease. AB - Immunocytochemical staining with monoclonal antibodies to the beta-protein on tissue sections which have been pretreated with formic acid is not only a very specific but also a highly sensitive method for the detection of amyloid deposits in the brains of Alzheimer's disease victims. We report here a spectrum of morphological appearance of the brain amyloid deposits which are one of the main histopathological correlates of this disorder. Deposits of the beta-protein are not only found in the well-known lesions [congophilic angiopathy and senile (neuritic) plaques] but are also seen under various morphological forms for which the word "plaques" does not appear an appropriate term: amyloid fibrils are found as large areas of diffuse infiltration of the neuropil, as ribbon-like infiltration in the subpial layer of the cerebral cortex, as granular deposits in the white matter, as diffuse deposits in the molecular layer of the cerebellum and the basal ganglia and as star-shaped deposits in the cerebellar Purkinje cell layer. The morphology of these deposits seems to depend on the cyto- and fibroarchitectonics of the brain region in which they are found, on the amount of amyloid deposited, and also on the type of staining technique used. It is only under specific circumstances that the deposition of amyloid in the neuropil is accompanied by the formation of paired helical filaments in nerve cell processes and their parent perikarya. In conclusion, our studies suggest that the extent of brain amyloidosis in Alzheimer's disease is much wider than so far appreciated. PMID- 2551123 TI - Microangiopathy of endoneurial vessels in hypoxemic chronic obstructive pulmonary disease (COPD). A quantitative ultrastructural study. AB - A peripheral neuropathy has been reported in patients with chronic respiratory insufficiency due to chronic obstructive pulmonary disease (COPD). It is mainly characterized by axonal degeneration, secondary demyelination and abnormal endoneurial vessels. The pathogenesis of these lesions remains obscure. To investigate whether relationships exist between neuritic and vascular lesions, a qualitative and quantitative ultrastructural study was performed on nerve biopsies in 13 patients with chronic respiratory insufficiency due to COPD, and in 9 normal controls without pulmonary lesions. A computer-assisted multiple regression analysis taking into account clinical, electrophysiological, biological and morphometric parameters was performed. Statistically significant differences in the endoneurial structure of microvessels were: (1) thickening of the basement membrane; (2) narrowing of the lumen; (3) mural pericytic debris deposits, occurring in the COPD group. In the latter, hypercapnia correlated positively with nerve fibers lesions (P = 0.03) and endothelial area (P = 0.03). No correlations were found between age and other parameters. These findings highlight the fact that the microangiopathy in peripheral nerves in patients with COPD may be diffuse and essentially due to hypoxia and reduction in blood flow, as in diabetic neuropathy. PMID- 2551125 TI - Kala-azar in a four-year-old child 18 months after brief exposure in Malta. AB - A four-year-old Danish boy developed kala-azar 18 months after a holiday in Malta. Splenectomy, with liver biopsy, was performed six months after onset of symptoms because of hypersplenism, and the diagnosis of kala-azar was only made four months later, when the histopathological specimens were reviewed. Previous bone marrow biopsies did not show Leishmania. Treatment with sodium stibogluconate was successful. The development of kala-azar after one week's stay in an endemic area stresses the importance of including this potentially fatal disease in the differential diagnosis of cases presenting with fever, splenomegaly, and pancytopenia. PMID- 2551124 TI - Immunodetection of the amyloid P component in Alzheimer's disease. AB - Amyloid P component (AP), a plasma constituent normally not found in brain parenchyma, has been immunohistochemically determined in brains from patients with Alzheimer's disease (AD). Tissue came from 11 clinically diagnosed and neuropathologically verified AD patients and from 6 normal aged controls. Positive labeling for AP was observed in amyloidotic blood vessels, senile plaques (SP) and neurofibrillary tangles (NFT). The immunoreactivity was specific for these AD-associated lesions and clearly revealed their morphological appearance. Affected blood vessels appeared to be mainly of the arteriolar type and were labeled abluminally in short segments. SP constituents such as amyloid fibrils, amyloid core and degenerative axonal and dendritic processes were positive for AP antiserum; the morphology and distribution of immunoreactive SP corresponded to previous descriptions. Labeling of NFT revealed the morphology of paired helical and straight filaments. In all cerebral areas studied, tangle bearing neurons were immunoreactive to AP antiserum, suggesting that AP is involved in early cellular development of NFT. Given the large molecular weight of AP (about 220,000), these results point to a potential impairment of the blood brain barrier in AD. Since AP is always present in systemic amyloidosis, its detection in cerebral amyloidosis associated with AD may suggest mechanisms common to the two disorders. PMID- 2551126 TI - Failure to deduce a peptide inhibitor of Na,K-ATPase from the gene coding for the catalytic alpha-subunit of Na,K-ATPase. AB - It has been proposed (Bost et al. 1985, Blalock & Bost 1986, Smith et al. 1987) that the DNA strand opposite the one coding, for example, for a hormone or a neurotransmitter codes for its receptor protein. This proposal was tested with regard to the possibility of deducing a peptide inhibitor of Na,K-ATPase from the known gene of the catalytic alpha-subunit of the enzyme. Three such peptides, deduced from presumed extracellular domains of the catalytic alpha-subunit, were tested in vitro and in vivo with regard to their effects on Na,K-ATPase activity. No effect on the enzyme could be demonstrated. PMID- 2551127 TI - Transport pathways for Na+ and Br- (Cl-) in noradrenaline-stimulated frog skin (Rana temporaria). AB - (1) Transport pathways for Na+ and Br- across noradrenaline-stimulated frog skin, in which the sodium channels in the apical membrane had been blocked with amiloride, were examined by pre-steady-state flux ratio experiments. (2) To analyse the experiments, equations are derived from which the fluxes through two parallel pathways can be determined if these differ with respect to the flux ratio as well as the mean passage time for the ion in question. (3) The fluxes of Na+ and Cl- were followed over a period of 3 h and it was found that about 1 h after addition of noradrenaline time-invariant fluxes could be achieved. In that period inward and outward tracer fluxes of Na+ and Br- (used as a substitute for Cl-) were followed during the build-up of isotope in the tissue until steady state fluxes were attained. (4) The experiments showed that when the Na channels of the capital cells are blocked the glands provide the major pathway for sodium transport across the skin. The efflux of Br- can be separated into two components. A fast one through the mitochondria-rich cells, which is the pathway for passive anion fluxes, and a slower one through the glands. No influx through the slow transport pathway could be detected. In some skins exchange of Br- through the mitochondria-rich cells is revealed. An anion exchange mechanism also appears to be present in the basolateral membrane of the gland cells. PMID- 2551128 TI - The effect of amiloride and benzimidazoleguanidine added to the inside medium on electrolyte pathways in the frog skin glands. AB - Serosal amiloride inhibits Na+, K+ and Cl- efflux and reduces short-circuit current and transepithelial conductance in noradrenaline-stimulated frog skin in which the sodium channels in the apical membrane are blocked by amiloride. BIG (benzimidazoleguanidine) inhibits Na+ and K+ efflux and reduces the short-circuit current. In some skins BIG decreased and in others it increased Cl- efflux and conductance. The variable response appears to be due not only to inhibition of salt extrusion from the gland cells but also to activation of Cl- transport through the mitochondria-rich cells, since it was shown that BIG could increase Cl- efflux in gland-free preparations. Different target mechanisms in the gland cells where the two substances may exert their effect are discussed. PMID- 2551129 TI - Density and functioning of human lymphocytic beta-adrenergic receptors during prolonged physical exercise. AB - In order to study the regulation of beta-adrenergic receptor number and function in response to prolonged physical effort, lymphocytic beta-adrenoceptor density (determined by (-)[125I]iodocyanopindolol binding), lymphocytic basal and isoproterenol-stimulated cyclic AMP (cAMP) production and concentrations of plasma catecholamines were measured before and during 3 h running exercise in eight healthy volunteers. A significant (P less than 0.01) increase of the lymphocytic beta-adrenoceptor density from 45 +/- 4 to 81 +/- 9 fmol mg-1 protein (mean +/- SEM) took place during the first hour of exercise. As the exercise was continued for up to 2.1-3 h, the receptor densities did not change significantly any more and remained elevated (72 +/- 9 fmol mg-1 protein) in comparison to the resting levels (P less than 0.02). The isoproterenol-stimulated cAMP production of the lymphocytes increased during the first hour of running from 190 +/- 36 to 269 +/- 56 pmol mg-1 protein (P less than 0.01) and returned to the resting level at the end of the exercise (182 +/- 38 pmol mg-1 protein). The mean levels of plasma catecholamines increased approximately sixfold during the first hour of exercise and remained elevated until the end of the running. This study demonstrates that the beta-adrenergic receptor system is activated in lymphocytes during prolonged aerobic physical exercise. This activated state becomes, however, attenuated within 2-3 h of exercise as indicated by a diminishing ability of beta-adrenoceptors to mediate catecholamine-induced cAMP production. PMID- 2551130 TI - Neurogenic contractions in caudal artery from young rats: receptor-receptor interaction and regional differences in effects of noradrenaline and ATP via alpha 1-, alpha 2- and P2x-receptors. PMID- 2551131 TI - Pholasin chemiluminescence detects mostly superoxide anion released from activated human neutrophils. AB - The bioluminescent oxygen metabolite indicator protein pholasin was characterized with respect to the type and location of reactive oxygen metabolites detected in suspensions of stimulated human neutrophils. Whereas pholasin detected reactive oxygen metabolites from neutrophil suspensions stimulated with soluble agents, particulate stimulants were apparently not effective triggering agents for pholasin-dependent neutrophil chemiluminescence. Neutrophils stimulated with fMet Leu-Phe (1 to 100 nmol/l) showed maximum pholasin-dependent chemiluminescence 45 to 60 s after stimulation. The time of maximum chemiluminescence was virtually independent of fMet-Leu-Phe concentration. In contrast, the time to reach maximum light emission increased from 60 s with 100 nmol/l phorbol ester to 295 s with 1 nmol/l phorbol ester. Significant inhibition of stimulated chemiluminescence was caused by both superoxide dismutase (20 micrograms/ml, 80% inhibition) and reduction of the oxygen concentration in the incubation medium to less than 0.5 mumol/l (95% inhibition). In contrast, the myeloperoxidase inhibitor sodium azide (0.1 mmol/l) afforded only 50% inhibition of the pholasin-dependent neutrophil chemiluminescence. Our results show that pholasin detects superoxide radicals released from cells stimulated by soluble stimulants but not intracellular oxidative activity elicited by particulate stimulants. PMID- 2551132 TI - Functional status of the pituitary-adrenal axis during treatment of RU486 for termination of early pregnancy in combination with PGs in Chinese women. AB - Six pregnant women (below 49 days of amenorrhea) desiring pregnancy termination received 600 mg RU486 orally in a single dosage and 1 mg PG-05 or ONO-802 as vaginal suppository 48 h afterward. Plasma cortisol, corticosterone, aldosterone and ACTH were monitored at -0.5, 1, 2, 4, 10, 24, 48, 72 h as well as during the first follow-up visit at the 8th day. Cortisol increased during the treatment and the values returned to the pretreatment level by the first follow-up visit. The changes of corticosterone were similar to those of cortisol. ACTH was also elevated within first 2 h and returned to the basal level at 48 h. There was no notable change in aldosterone during the treatment. The results suggest that a compensation of the pituitary-adrenal function occurred during the treatment, which is probably associated with a decreased negative feedback mechanism of cortisol due to the antiglucocorticosteroid activity of RU486 at the hypothalamus level. PMID- 2551133 TI - Multicomponent analysis of near-infrared spectra of anesthetized rat head: (II). Quantitative multivariate analysis of hemoglobin and cytochrome oxidase by non negative least squares method. AB - We developed a method for quantitating absolute value of Hb oxygenation and that of redox state of cyt. aa3 on the basis of multiple regression analysis of near infrared spectrophotometric data. Flattening of the spectrum of Hb in both RBC suspension and the brain in situ was observed. This phenomenon was explained by localization of Hb within RBC for in vitro flattening, and by localization of RBC within the vessels as well as deformation of RBC in the microcirculation in vivo. Under resting air-breathing conditions, So2 and redox state of cyt. aa3 were estimated to be 50-70% and over 90%, respectively. Increase in FIO2 up to 3 ATA O2 caused stepwise increase in So2, whereas cyt. aa3 was fully oxidized when the animal inspired O2 under 1 ATA. Problems which must be solved for more accurate estimation of absolute values of So2 and redox state of cyt. aa3 were described and discussed. PMID- 2551134 TI - Multicomponent analysis of near-infrared spectra of anesthetized rat head: (I). Estimation of component spectra by principal component analysis. AB - By measuring precise NIR spectra of anesthetized rat head, we determined principal components included in changes of the NIR spectra caused by acute hypoxia. There was apparent delay in reduction of cyt. aa3 as compared with Hb deoxygenation in the hypoxic period. With regard to the eigenvalues determined by principal component analysis, principal components in the NIR spectral changes caused by acute hypoxia were Hb and cyt. aa3, and contribution of the remaining biological materials to the changes in NIR spectra caused by acute hypoxia was considered to be negligible in quantitative multivariate analysis of Hb and cyt. aa3 in situ. PMID- 2551135 TI - A CCD spectrophotometer to quantitate the concentration of chromophores in living tissue utilising the absorption peak of water at 975 nm. PMID- 2551136 TI - Quantitation of pathlength in optical spectroscopy. PMID- 2551137 TI - The oxygen dependence of the energy state of cardiac tissue: 31P-NMR and optical measurement of myoglobin in perfused rat heart. AB - The relationship between the energy state and intracellular oxygen concentration was established with cardiac tissue, where the former could be monitored by 31P NMR and the latter by optical method for myoglobin absorption. The ratio of creatinephosphate to inorganic phosphate, PCr/Pi, started to fall at higher intracellular oxygen concentration than oxygen consumption (10IM). The pH shift measured by frequency shift of Pi paralleled the increase of lactate-release, of which half maximum was approximately 3IM. The critical oxygen concentrations differed nearly 5-fold when determined by energy state and oxygen consumption. PMID- 2551139 TI - Effects of leukocyte-derived oxidants on sarcolemmal Na,K,ATP-ase and calcium transport. AB - Our study demonstrated that the Na,K,ATPase activity and ouabain binding sites were reduced by oxidants. Sarcolemmal calcium transport was also inhibited by hydrogen peroxide and HOC1. The action of HOC1 on the sarcolemmal functions was 2 3 orders of magnitude more powerful than that of hydrogen peroxide. Effects of hydrogen peroxide consisted of two components, i.e., the first, highly sensitive one, most probably mediated by Fe-catalyzed, site-specific free radical formation, and the second, less potent action by (high concentrations of) hydrogen peroxide. Finally, very low concentrations of hydrogen peroxide potentiated Na,K,ATPase activities when assayed using myocytes. PMID- 2551138 TI - The effects of temperature and buffer concentration on the metabolism of human atrial appendages measured by 31P and 1H NMR. AB - In this study we measured changes in intracellular ATP and pH together with lactate production in isolated ischemic human atrial tissue. The measurements were made using 31P and 1H NMR. ATP preservation is improved as temperature is reduced from 20 degrees C to 1 degree C because of a progressive decrease in energy demand. At a constant temperature (12 degrees C), ATP preservation is improved by increasing the extracellular buffer capacity with PIPES buffer at concentrations up to 100 mM. Under these conditions, energy demand appears to increase but the ATP level is kept relatively constant for periods of 10 hours or longer. This appears to be due to a tighter regulation between supply and demand in which glycolysis is driven faster at relatively lower ADP and Pi levels. This tight regulation may be attributed to the better maintenance of intracellular pH. PMID- 2551140 TI - Near infrared quadruple wavelength spectrophotometry of the rat head. AB - A quadruple wavelength method to monitor the changes in concentration of oxygenated and deoxygenated hemoglobin and the redox state of cytochrome oxidase within a living tissue is presented. The expected advantages of this technique over the triple wavelength method are (i) that it can compensate for the light scattering change of tissue itself or for instabilities of light source and photomultiplier, (ii) that it can treat the optical properties of the red blood cell in a tissue in the same way as in an in vitro model system, and (iii) that it requires no estimation of the absorption coefficient of cytochrome oxidase in situ. PMID- 2551141 TI - The oxygen dependency of the redox state of heme and copper in cytochrome oxidase in vitro. AB - The oxidation-reduction state of cytochrome oxidase in isolated mitochondria at low oxygen concentrations was measured by the use of leghemoglobin as an oxygen indicator. P50 a + a3 varied with energy state as well as the respiratory rate. In contrast to heme a + a3, copper was slower to reduce than heme a + a3. The P50Cu of 8 x 10(-8)M in State 4 and 7.4 x 10(-8)M in State 3 was independent of both the energy state and the respiratory rate. PMID- 2551142 TI - Reversal of carbon monoxide-cytochrome c oxidase binding by hyperbaric oxygen in vivo. AB - Cytochrome a,a3 redox state of the parietal cortex of pentobarbital anesthetized rats was continuously monitored through intact skull with four wavelength differential spectrophotometry during exposure to 90% O2 plus either 1.0 or 0.5% CO at 1 and 3 (ATA). The formation of HbCO was monitored in the brain by absorbance differences between 569 and 586 nm and correlated positively in graded 0.25 to 1% CO exposures with measured HbCO levels. Exposure to 90% O2, 1% or 0.5% CO (balance N2) decreased mean arterial pressure (MAP), calculated arterial O2 content and cytochrome a,a3 oxidation measured at 605 nm relative to 620 nm while HbCO rose. After compression to 3 ATA, rats breathing CO mixtures increased MAP and O2 content with reoxidation of cytochrome a,a3 while HbCO remained constant. Further treatment of both groups with 90% O2 at 3 ATA recovered the above parameters to at least control values except small persistent elevations of HbCO. Difference spectra recorded from 568 to 620 nm in parallel experiments showed twin absorbance peaks at 588 to 592 nm and 600 to 605 nm in response to CO. These absorbance maxima were consistent with formation of the cytochrome a3-CO complex and cytochrome a reduction respectively. These studies indicate that CO binds to reduced cytochrome a3 in blood circulated rat cortex in CO hypoxia and this effect can be reversed by increasing dissolved arterial O2 content at 3 ATA. PMID- 2551143 TI - Oxygen diffusion through mitochondrial membranes. AB - The effect of the mitochondrial membrane on the oxygen supply to the interior of mitochondria was analyzed with a cylinder model of diffusion. This estimation is based on the assumption that cytochrome a,a3 is distributed only on the inner surface of the mitochondrial inner membrane. The diffusion coefficient in the mitochondrial membrane was approximated from the fluorescently-determined viscosity of rat mitochondrial membrane. A pico-second time-resolved fluorometer at 37 degrees C gave values of 43.8 cp for intact mitochondria and 51.4 cp after phospholipase A2 treatment. Using the mean oxygen consumption rate of 10 ml O2/100 g tissue/sec in beating heart, oxygen gradients of 3.9 and 4.6 nmol was predicted across the intact and phospholipase-A2 treated mitochondrial membranes, respectively. The increased oxygen consumption during systole will yield oxygen gradients of 11.6 and 13.7 nmol. These gradients were much larger than the values estimated in a hypothetical case using the diffusion coefficient for the mitochondrial membrane of 1.5 x 10(-5) cm2/sec. The predicted oxygen gradient suggests a non-uniform distribution of oxygen in the myocardial cell and may be of importance in understanding the relationship between oxygen supply and myocardial function in hypoxia. Phospholipase A2, which is known to be activated in ischemia, destroys the microstructure of myocardial cells, seems deleterious to oxygen transport to cytochrome a,a3. PMID- 2551144 TI - Behavior of the copper band of cytochrome c oxidase in rat brain during FC-43-for blood substitution. AB - FC-43-for-blood substitution experiments were conducted in anesthetized rats to evaluate NIR spectroscopic responses by living brain to exchange transfusion at very low hematocrits. The NIR responses to the exchange process also assess the ability of multiwavelength algorithms to measure independent changes in the relative amounts of Hb, tHbO2 and oxidized cytochrome a,a3 in the tissue. These data indicate that FC-43 circulation at high FIO2 (1.00) delivers sufficient O2 to the rat brain to maintain the oxidation state of the mitochondrial oxidase near pre-exchange values. The results also confirm the ability of four wavelength algorithms to distinguish changes in the oxidized copper band of cytochrome a,a3 from changes in absorption by Hb and HbO2. PMID- 2551145 TI - Effects of canrenone on Na+,K+ ATPase activity, arterial pressure and plasma potassium concentration in uremic hemodialyzed patients. PMID- 2551146 TI - The leukocyte integrins. PMID- 2551148 TI - A perspective of Epstein-Barr virus diseases. PMID- 2551147 TI - Structure and function of the complement receptors, CR1 (CD35) and CR2 (CD21). PMID- 2551149 TI - Effect of auranofin on eicosanoids and protein kinase C in human neutrophils. AB - Auranofin (AF), a lipophilic chrysotherapeutic agent, was investigated for its effect on the formation of lipoxygenase products and the activity of protein kinase C in human neutrophils. We have previously shown that inhibition of LTB4 formation by 5-lipoxygenase (5-LO) inhibitors is intimately associated with a marked increased in 15-HETE in excess of arachidonic acid. The calcium- and phospholipid-dependent protein kinase, protein kinase C, is activated in FMLP- and A23187-stimulated neutrophils, is hypothesized to stimulate superoxide generation, and plays an essential role in eicosanoid production. AF dose dependently inhibited the generation of leukotriene B4(LTB4) in FMLP-stimulated neutrophils, the ID50 was approximately 4.5 micrograms/ml. Unlike known 5-LO inhibitors, AF did not enhance the production of 15-HETE. In neutrophils stimulated with the calcium ionophore, A23187, AF did not inhibit the generation of LTB4 nor did AF change the 15-HETE levels. AF inhibited superoxide generation in FMLP-stimulated neutrophils dose-dependently, but did not change the activation of protein kinase C in the cells. We therefore conclude, that AF inhibition of LTB4 production in neutrophils is different from 5-lipoxygenase inhibitors and is elicited at a step distal to protein kinase C activation. PMID- 2551150 TI - Fish oil diet rich in eicosapentaenoic acid increases bleeding time in the rat by interaction with sympathetic transmitters. AB - The influence of a diet supplemented with MaxEPA (a fish oil concentrate, rich in eicosapentaenoic acid = EPA) on bleeding time in small mesenteric arteries of the rat and on formation of thromboxane B2 (TXB2) by ADP-stimulated platelets was investigated. Since EPA has been found to antagonize noradrenaline (NA) - induced vasoconstriction, the influences on bleeding time of the alpha-receptor antagonist phentolamine and of the adrenergic neuron blocking agent guanethidine alone as well as during dietary supplementation with MaxEPA were studied. Bleeding time slowly increased during the MaxEPA diet over a period of 12 weeks resulting in a final prolongation of about 75%. Indomethacin which dose dependently increased bleeding time in control rats, further increased the already prolonged bleeding time in rats on the MaxEPA diet. Guanethidine prolonged bleeding time strongly in control animals but caused no further increase in those on the MaxEPA diet. Indomethacin also further increased the bleeding time prolonged by guanethidine, just as it did in animals on MaxEPA diet. In contrast to guanethidine, phentolamine prolonged bleeding time only slightly. The ability of platelets from rats on the diet to generate TXB2 was reduced to 59.5% but was more drastically reduced (to 26.7%) from rats pretreated with indomethacin. The results indicate that MaxEPA feeding prolongs bleeding time by a mechanism other than that of indomethacin. We speculate that EPA prolongs bleeding time rather by antagonizing the vasoconstrictor effect of sympathetic transmitters at the site of the injured vessel than by a reduced formation of TXA2. PMID- 2551151 TI - Effects of PAF and PAF antagonists on the shape of venous endothelial cells in vitro. AB - Three platelet-activating factor (PAF) antagonists were tested for their ability to prevent or reduce PAF-induced shape changes of large vein endothelial cells in vitro. BN52021 had a significant protective action at concentrations of 1 microM and 0.1 microM, but at 100 microM had a damaging effect of its own. CV3988 (0.1 microM and 1 microM) and L652,731 (20 microM) did not reduce the responses to PAF, and at higher concentrations (CV3988 10 microM and 100 microM, L652,731 100 microM) both compounds alone caused significant changes of shape. BN52021 (0.1 microM) was also effective against leukotriene (LT) C4, at 1 microM against bradykinin and LTE4, and at 10 microM against LTD4 and the calcium ionophore A23187. BN52021 (10 microM) was ineffective against shape changes induced by histamine, prostaglandin (PG) E2 and lysophosphatidylcholine (LPC). Neither indomethacin (100 microM) nor verapamil (20 microM) altered the response to PAF. Using electron spin resonance (ESR) spectrometry it was shown that the damaging effects of LPC and CV3988 may be due partly to their detergent properties. It is suggested that the mechanism by which PAF alters the shape of large vein endothelial cells is primarily receptor mediated. PMID- 2551152 TI - Orally administered tolfenamic acid inhibits leukotriene synthesis in isolated human peripheral polymorphonuclear leukocytes. AB - Special interest has been focused on the development of dual inhibitors of the cyclo-oxygenase and lipoxygenase pathways of arachidonic acid metabolism. In contrast to other classic NSAIDs, some fenamates in clinically achievable concentrations have been shown to inhibit synthesis of 5-lipoxygenase products in vitro. In the present work, we studied the effect of orally administered tolfenamic acid (600 mg) on Ca ionophore A 23187 -induced leukotriene synthesis in isolated human polymorphonuclear leukocytes. Leukotriene production was reduced in all 14 subjects studied, the mean inhibition of LTB4 synthesis being 16 +/- 3% and that of LTC4 33 +/ 7%. The inhibition correlated positively with serum acid concentrations. We suggest that inhibition of leukotriene synthesis is an additional mechanism of the anti-inflammatory, antimigraine and antidysmenorrhoeic effects of tolfenamic acid, and a possible explanation for its rare gastric and bronchoconstrictive side-effects. PMID- 2551154 TI - Epidemiological studies of piglet diarrhoea in intensively managed Danish sow herds. IV. Pathogenicity of porcine rotavirus. AB - Colostrum-deprived piglets inoculated with rotavirus 24 h after birth developed a profuse diarrhoea that spread to non-inoculated, colostrum-deprived litter mates and, occassionally, to colostrum-fed piglets. Case fatality rates in these 3 categories of piglets were 63.2%, 35.7% and 8.3%, respectively. Surviving piglets recovered in 1-2 weeks, but shedded virus via the faeces for up to 3 weeks p.i. The D-xylose test revealed severe malabsorption, with extremely flat absorption curves for up to 3-4 weeks p.i. Malabsorption was more marked in piglets with a long-lasting faecal virus excretion than in piglets where virus disappeared from the faeces within 10 days p.i. Infected piglets (colostrum-fed and colostrum deprived) had decreased weight gains and were 5 days older at a bodyweight of 25 kg than non-inoculated controls. It is concluded that rotavirus is probably of significance in diarrhoeal syndromes in suckling piglets, alone or in combination with E. coli or other pathogens. PMID- 2551155 TI - Properties of lactase produced by enteropathogenic Escherichia coli from diarrhoeic children. AB - The quantity of lactase produced by enteropathogenic Escherichia coli (EPEC) was significantly higher (P less than 0.01) than that produced by non-EPEC. The enzyme production was induced by lactose but repressed by glucose and galactose. The lactase from EPEC which was partially purified by ammonium sulphate precipitation and gel permeation chromatography had a molecular weight of 56 kD and apparent Km of approximately 2.78 mM for lactose. The lactase exhibited optimum activity at pH 7.0 at 40 degree C and was stimulated by Mg2+, Mn2+, Na+ and inhibited by Ba2+, Ca+, Cu2+, EDTA, iodo acetic acid (IAA) and Hg2+ and U2+ ions. The higher production of lactase by EPEC may be linked to its pathogenic role in childhood diarrhoea. PMID- 2551153 TI - Neutrophil enzyme activities in carrageenan-induced inflammation in rats. AB - During the course of carrageenan-induced inflammation in rats major changes were observed in the activities of neutrophil granule enzymes. The activities of three enzymes--gelatinase, collagenase and beta-glucuronidase, the markers of three different types of granules, have been measured and compared to those of a control group of animals. Total collagenase and gelatinase activities of control rats were 59.4 +/- 3.6 (mean +/- SD) and 23.0 +/- 2.9 units/mg protein, respectively. Significantly reduced levels of both collagenase (35.6 +/- 2.5 units) (p less than 0.05) and gelatinase (7.1 +/- 0.7 units) (p less than 0.001) were measured in the blood neutrophils of inflamed rats; and the collagenase activity of neutrophils derived from inflamed pleural exudate was also significantly decreased to a level of 19.7 +/- 1.8 units/mg protein (p less than 0.01). However, the gelatinase activity of exudate neutrophils did not differ from that of blood cells of inflamed rats. In contrast, no change was found for the beta-glucuronidase activity in blood neutrophils of control and inflamed rats. These observations support the concept that during the inflammatory response in rats, neutrophils in the circulation may become activated as judged by the extracellular secretion of collagenase and gelatinase. Therefore, neutrophils accumulating in acute inflammatory lesions contain decreased levels of collagenolytic enzymes and the significance of this observation is discussed. PMID- 2551156 TI - Serum immunoglobulin concentration in Nigerian infants. AB - Serum immunoglobulin (Ig) G, M and A were determined at 3-monthly intervals during the first year of life in 35 healthy Nigerian infants. The neonatal IgG values were high, but dropped rapidly by 3 months to about 37% of the neonatal value, and thereafter rose steadily. The neonatal IgM values dropped slightly (10%) by 3 months and then rose steadily to reach a level above the neonatal value after 1 year. IgA was not detected in most of the children during the neonatal period, but where it was detected and was measurable, the values were very low and then rose steadily until the age of 1 year. There was a suggestion that the pattern of immunoglobulin in infancy might be influenced by the level of maternal education. PMID- 2551157 TI - Transient ischaemic attacks: a retrospective study of 43 cases. AB - Forty-three Caucasians with transient ischaemic attacks (TIAs), all age- and sex match controlled, were retrospectively studied. There was no difference in sex distribution. The peak age for TIAs was 55-64. The haematocrit and cholesterol levels of men were significantly elevated (0.01 greater than P greater than 0.001 for haematocrit, and 0.01 greater than P greater than 0.001 for cholesterol). Six patients were controlled hypertensives but in general there was no significant difference in the blood pressures of patients and controls. The risk factors for TIAs and strokes are discussed. PMID- 2551158 TI - Measurement of sialyl-transferase activity in isolated colonic mucosal cells of the rat. AB - An assay for sialyl-transferase activity in isolated rat colonic mucosal cells has been developed. Total and specific activity with asialobovine sub-mandibular gland glycoprotein and endogenous substrates was approximately two-fold higher in the proximal, relative to the distal, colon. These activities were similar when asialo-alpha-acid glycoprotein was used as substrate. Endogenous activity was approximately 10-fold lower than with exogenous substrates in both proximal and distal colonic segments. Analysis of total and specific sialyl-transferase activity up to 7 weeks after jejunoileal bypass (JIB) and sham operation showed a marked increase at the first week, decreasing toward normal by week 7. Similar differences between proximal and distal colon, and with the type of substrate, were found with both groups of operated animals. Histochemical analysis showed small elevation after 1 week for sialo and sulphomucins after the JIB operation only. PMID- 2551159 TI - Intestinal permeability as a measure of small intestinal mucosal integrity: correlation with jejunal biopsy. AB - The permeability of the small intestine was measured and jejunal biopsy performed in 39 children with gastrointestinal disorders. Intestinal permeability was measured using orally administered mannitol and lactulose as probe molecules in an isotonic solution (274 mOs/l), and the results were expressed as the ratio of the urinary excretion of the two sugars over 5 h. Urine samples were analysed for mannitol and lactulose content by high performance liquid chromatography. Children with small intestinal mucosal damage, irrespective of the cause, had a significantly lower (P less than 0.001) mannitol excretion (mean recovery 1.21% of ingested dose) than those with a normal mucosa (mean recovery 5.3%), while lactulose excretion did not differ (P greater than 0.05). The mean value of the lactulose:mannitol urinary excretion ratios was significantly higher (P less than 0.001) in subjects with an abnormal mucosa (0.98) compared to those with a normal mucosa (0.2). Using the mean plus two standard deviations of the normal mucosa group to define the upper limit of normal, all lactulose: mannitol excretion ratios from the abnormal mucosa group were above this limit. The results of this study show that the sugar permeability test is a sensitive, non-invasive screening test for jejunal mucosal damage in children and shows good correlation with jejunal biopsy results. PMID- 2551160 TI - Five years experience of haemodialysis at the Lagos University Teaching Hospital- November 1981 to November 1986. AB - The Dialysis Centre at the Lagos University Teaching Hospital became operational in November 1981 and caters for acute haemodialysis, chronic maintenance haemodialysis and continuous arteriovenous haemofiltration. In the past 5 years, over 600 patients had presented out of whom 245 could be accommodated within the realities of available facilities and patients' financial status. Of the 245 patients, 25 were discharged against medical advice and five were transferred to hospitals abroad but did not survive. There were 117 patients in end-stage renal failure (ESRF), 75 males, 42 females, ratio M:F 1.8:1, age range 13-69 years, mean 37.5. There were 51 males and 47 females in acute renal failure (ARF), ratio 1.1:1, age range 13-76 years, mean age 32.3 (Table 1). All patients in ESRF had moderate to severe hypertension (diastolic pressure of greater than or equal to 120 mmHg or 22.1 kPa) and a creatinine clearance of less than or equal to 5 ml/min and about 75% had established cardiac decompensation. Full pertinent investigations were precluded or contra-indicated in most patients in ESRF because of late presentation. In only 13 patients was renal biopsy performed and the pathohistologies were end stage renal disease (8), chronic glomerulonephritis (4) and glomerulosclerosis (1). In ARF the cause of the renal damage was multifactorial in 66.7%, with sepsis being the direct cause of death in 60.0%. The commonest conditions were septicaemia (61.4%), nephrotoxin (17.2%), trauma (31.3%), septic abortion (33.3%) and toxaemia of pregnancy (29.0%) (Table 2). The dialysis associated complications which were encountered included shunt infection (7%), burst membrane (9%), suspected pyrogen reaction (5.6%) and femoral vein perforation (0.9%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551161 TI - Neuropsychiatric disorders in Nigerians: 1914 consecutive new patients seen in 1 year. AB - The socio-demographic attributes and the different diagnostic categories of patients attending Aro Neuropsychiatric Hospital for the first time over a 1-year period are presented. There was an overall preponderance of males but more females than males suffered from depression. Factors which distinguished patients with anxiety neurosis from those with neurotic and endogenous depression are identified. Two patients who suffered from obsessional neurosis, commonly regarded as rare in the black Africans, are described. Heredodegenerative diseases of the nervous system are rare in the Africans and one patient with hereditary spinocerebellar degeneration is described. Eclampsia was a probable predisposing factor for epilepsy in four women. PMID- 2551162 TI - Detection and determination of salicylic acid impurity in aspirin tablet formulations' by high performance liquid chromatography. AB - Salicylic acid is a major hydrolytic degradation product of aspirin, responsible especially for gastric irritation during oral aspirin administration. This impurity was investigated in 12 different brands of aspirin formulation readily available in our locality. A simple, rapid and sensitive high performance liquid chromatographic method was adopted for this investigation. The mobile phase was methanol/water (20/80, v/v) adjusted to pH 2.5 with phosphoric acid and was run on a 50 mm reversed-phase column monitored at 240 nm. The limit of detection for salicylic acid was 5 ng. Only three of these formulations showed the presence of salicylic acid impurity and all these contained salicylic acid in excess of the USP 1980 limit of 0.3% salicylic acid per tablet. PMID- 2551163 TI - Hodgkin's disease in siblings: a case report. AB - Two male siblings (ages 12 and 16 years) presenting with Hodgkin's disease are reported. They were both diagnosed as stage IVB with identical histological type- lymphocyte depleted. The presence of identical sex, shared environment and the closeness of the time of onset suggested a combination of both environmental and genetic factors in the aetiology of the disease in these siblings. PMID- 2551164 TI - Prevalence of cholelithiasis in Nigerians with sickle cell disease. AB - Gall bladder ultrasonography was performed on 157 fasting patients (mainly adults) with sickle cell disease (133 SS and 24 SC) with a view to establishing the prevalence of cholelithiasis in Nigerian 'sicklers'. There were 82 females and 75 males whose ages ranged from 9 to 60 years (mean 34.5). Gallstones were demonstrated in 38 patients, giving an overall prevalence of 24.2% (22.6% in SS and 33.3% in SC). Their ages ranged from 18 to 56 years (mean 37.0) in HbS + C patients and 10-34 (mean 22.0) in SS patients. All the patients except one were asymptomatic. Autopsy confirmed gallstones in two SS patients who died of unrelated problems. The higher prevalence obtained in this study (24.2%) compared with the prevalence (less than 10%) in earlier studies from Africa could be due to the predominantly adult age group screened and the greater sensitivity of cholecystosonography. PMID- 2551165 TI - Plasma lipid profiles in relation to diabetic control in Nigerians. AB - Plasma lipid profiles--total cholesterol (TC), LDL-cholesterol, HDL-cholesterol, triglycerides and phospholipids--were studied in relation to two parameters of diabetic control (fasting blood sugar (FBS) for short-term control and glycosylated haemoglobin (HBA1C) for long-term control) in 46 diabetic patients (22 insulin-dependent (IDDM) and 24 non-insulin dependent (NIDDM] and 22 non diabetic control subjects. We confirmed the positive correlation between FBS and HBA1C. All diabetic patients had significantly higher triglyceride levels (P less than 0.05) than controls, which were not influenced by degree of glycaemic control. NIDDM patients tended to have higher than normal TC levels (P less than 0.05). In IDDM, TC level was positively correlated with HBA1C (r = 0.37, P less than 0.05), and negative correlations were established between FBS and HDL cholesterol (r = -0.46, P less than 0.02) and the HDL-cholesterol:TC ratio (r = 0.49, P less than 0.01), suggesting an increased atherogenic risk with poorer diabetic control. It is concluded that lipoprotein abnormalities exist in Nigerian diabetics, though not as consistently as in Caucasians. The differences may be due to, among other factors, differences in genetic make-up, diet (typical African diet being rich in plant fibre and poor in cholesterogenic nutrients) and aetiology of the diabetic state (tropical diabetes being highly heterogeneous and now thought to be linked to malnutrition). PMID- 2551166 TI - Immunity in malaria: II. Heterophile and malarial antibodies in acute Plasmodium falciparum infection. AB - The sera of 55 Nigerian children (30 malarious and 25 healthy) were analysed for heterophile antibodies against normal sheep erythrocytes by the passive haemagglutination technique. Fluorescent antibodies to Plasmodium falciparum were quantified by the indirect immunofluorescence method while the three major immunoglobulins (IgG, IgA and IgM) were estimated by the radial immunodiffusion technique. An increasing age gradient was demonstrated in the heterophile antibody titres within the malarious and control groups, but there was no significant difference in the levels of immunoglobulins and malarial antibodies between the two groups. An indication of higher malarial antibody titre was only observed in the malarious group, particularly in late childhood. These results show an increasing level of heterophile antibodies with age. It is concluded that malarial antigens may play a contributory, but not a dominant role in the acquisition of heterophile antibodies. There is also a need to define the exact serum factors (antibody or non-antibody) which are associated with clinical immunity to malaria in Nigerian children. PMID- 2551167 TI - Alpha 2-adrenoceptors and endothelium-derived relaxing factor. AB - The endothelium can release potent vasodilator substances, in particular prostacyclin and endothelium-derived relaxing factor. The triggers for the release of endothelium-derived relaxing factor include increases in levels of shear stress, neurotransmitters, autacoids, platelet products, and hormones. The endothelium-dependent response to catecholamines involves stimulation of alpha 2 adrenoceptors on the endothelial cells. Indeed, in a number of blood vessels, selective alpha 2-adrenergic agonists cause endothelium-dependent relaxations. These are seen most typically in blood vessels with long-term exposure to high flows and high partial pressures of oxygen. In addition to the release of endothelium-derived relaxing factor, alpha 2-adrenergic agonists can stimulate postjunctional (postsynaptic) alpha 2-adrenoceptors on vascular smooth muscles. These receptors, which are more abundant in hypertensive blood vessels, activate the contractile process. However, the alpha 2-adrenergic vasoconstrictors act as partial agonists (with a limited receptor reserve) and hence their vasoconstrictor response is very sensitive to functional antagonists such as endothelium-derived relaxing factor. Thus, the presence of endothelial cells can blunt the vasoconstrictor response to these substances not only because of an augmented release of endothelium-derived relaxing factor but also because the vasoconstriction that they induce is particularly susceptible to the inhibitory effect of the factor. PMID- 2551168 TI - A clinical-pathologic study of mixed mullerian tumors of the uterus over a 16 year period--the Medical College of Georgia experience. AB - From 1972 through 1987, 40 patients at the Medical College of Georgia were assessed with a diagnosis of mixed mullerian tumors which constituted 3.5% of all female genital tract malignancies. The mean patient age was 65.3 years; 60% of the patients had stage I disease. Of those patients with clinical stage I disease, more advanced disease was found at celiotomy in 33%. Retroperitoneal nodes were positive for malignancy in 35% of 20 patients who underwent node sampling. Cell washings were positive in 12%, and 9% had omental metastases. Forty-seven percent had homologous tumors; 53% of tumors were heterologous. Relatively poor prognosis was associated with large tumor volume, vascular invasion, nodal metastases, and disease outside the uterus. The overall survival rate was 32%; 14 of 24 patients with stage I disease are dead of disease. Of patients dead of disease, 92.5% had distant metastases. Adjuvant therapy with a combination of VP-16, cisplatin, and irradiation was beneficial in four high-risk patients. PMID- 2551170 TI - Primary and secondary cutaneous Ki-1+ (CD30+) anaplastic large cell lymphomas. Morphologic, immunohistologic, and clinical-characteristics. AB - Skin biopsies were collected from 15 patients with cutaneous tumors morphologically similar to Ki-1+ anaplastic large cell (ALC) lymphoma of the lymph nodes, including Ki-1+ ALC lymphoma of childhood. The histology and immunophenotype of these cutaneous tumors are reported and follow-up data on the patients are given. The tumorous infiltrates were composed of large, sometimes very bizarre cells with one nucleolus or multiple nucleoli. All tumor cells expressed the lymphoid cell activation antigen Ki-1 (CD30) in conjunction with CD25 and the beta-chain of the T cell receptor. In 11 patients, Ki-1+ cutaneous tumors developed primarily in the skin. In nine patients, these were restricted to the skin in follow-up periods ranging from 3 months to 6 years. Two patients developed lymph node involvement after 2 months and 2 years, indicating the spreading potential of these cutaneous tumors. Morphologic and immunophenotypical identity of the atypical cells found in primary cutaneous ALC lymphoma, in regressing atypical histiocytosis (RAH), and in lymphomatoid papulosis (LyP) of type A, together with the protracted clinical course in all three conditions, suggests that primary cutaneous ALC lymphoma, RAH, and LyP of type A represent clinical variants of the same lymphoma entity. Secondary development of Ki-1+ ALC skin tumors was observed in two patients with cutaneous T cell lymphomas of mycosis fungoides type. These secondary Ki-1+ ALC lymphomas of the skin showed rapid systemic progression similar to the primary lymphonodal Ki-1+ ALC lymphomas. Concomitant or subsequent occurrence of Ki-1+ ALC tumors in cutaneous T cell lymphomas thus may be a bad prognostic sign. PMID- 2551169 TI - Transplacental induction of peripheral nervous tumor in the Syrian golden hamster by N-nitroso-N-ethylurea. A new animal model for von Recklinghausen's neurofibromatosis. AB - Multiple peripheral nervous tumors were induced in 45 of 60 (75.0%) Syrian golden hamsters by transplacental administration of N-ethyl-N-nitro-sourea. Moreover, melanomas, pheochromocytomas, and Wilms' tumors developed in six (10.0%), three (5.0%), and 13 (21.7%) animals, respectively. The histologic, immunohistochemical, and electron microscopic findings of the peripheral nervous tumors were similar to those of human neurofibroma, and their growth pattern and distribution resembled those of human von Recklinghausen's neurofibromatosis (VRNF). The occurrence of melanoma, pheochromocytoma, and proliferative foci of melanin-containing cells in neurofibroma suggests that the targets of ENU in hamsters are the neural crest-derived cells. With its high incidence of Wilms' tumor, the hamster with ENU-induced tumors is considered to be a good animal model for human neurocristopathy, including VRNF. PMID- 2551171 TI - Unique male mammary tumors developing in the inbred soft-furred field rats Millardia meltada. AB - Inbred lines of the soft-furred rat, Millardia meltada, were studied with special reference to spontaneous male mammary tumors. Adult males had the hyperplastic, pigmented inguinal mammary tissues and frequently developed bilateral mammary tumors. The tumors, no longer pigmented, were histologically well-differentiated adenocarcinomas associated with the myoepithelial cells and showed a variety of growth patterns depending on the stage of progression. They were transplantable to male, but not to female or castrated male nude mice. A significant number of androgen as well as estrogen binding sites were demonstrated in the nuclei of the transplanted tumor cells by in situ autoradiography. Neither virus particle nor mouse mammary tumor virus-specific antigen expression was detected in the tumor cells by electron microscopic and immunohistochemical studies. Normal females were generally free of pigment and tumor, although the androgen treatment induced marked pigmentation (deposit of nonfluorescent lipofuscin and hemosiderin) and hyperplasia of the female mammary epithelial cells, without a tumor. Possible relationships between pigmentation and tumor development were considered. PMID- 2551172 TI - Dual lineage of astrocytomas. AB - Experimental observations suggest that type 1 and type 2 astrocytes have distinct lineages distinguished, respectively, by the absence or presence of the A2B5 antigen. To investigate the hypothesis that the strikingly different biological behaviors of low-grade and high-grade astrocytomas might be related to cell lineage, 38 astrocytomas (grades I through IV), were examined for the presence of the A2B5 antigen, glial fibrillary acidic protein (GFAP), and galactocerebroside (GC), a marker for oligodendrocytes. Twenty-nine of the tumors (76%) were composed of moderate or high densities of GFAP+ and unlabeled cells, and were devoid of A2B5+ and GC+ neoplastic cells. Only four tumors had high densities (75% to 100%) of A2B5+ cells, three of which were grade I tumors; two tumors, also Grade I, contained moderate densities (20% to 25%) of GC+ cells that had cytologic features of astrocytoma or oligodendroglioma. The findings suggest that most cerebral astrocytomas lack the A2B5 antigen and thus may represent neoplasia along the type 1 astrocyte lineage. In contrast, A2B5+ lineage among astrocytomas, ie, neoplasia with differentiation toward type 2 astrocytes and GC+ oligodendrocytes, is less common and may be correlated with prognostically favorable histopathologic features. PMID- 2551173 TI - Terminal cisternae of denervated rabbit skeletal muscle: alterations of functional properties of Ca2+ release channels. AB - Terminal cisternae (TC) of skeletal muscle represent the specialized compartment from which Ca2+ is released into the myoplasm after a propagated action potential. In this study we have investigated the morphology, protein composition, and Ca2+ release properties of TC isolated from rabbit gastrocnemius muscle 2 wk after nerve sectioning. Thin-section electron microscopy showed that TC vesicles from denervated muscle were enriched in calsequestrin (CS) and contained a larger fraction of the junctional sarcoplasmic reticulum (SR), as judged by membrane profiles with morphologically intact feet structures. Accordingly, the yield of junctional SR from denervated muscle was twice that of control muscle, and the protein pattern of TC vesicles exhibited an increase in junctional protein components, e.g., CS and the 350-kDa protein. The larger content of the 350-kDa protein, or ryanodine receptor (F.A. Lai, H. Erickson, E. Rousseau, Q.-Y. Liu, and G. Meissner, Nature Lond. 331: 315-319, 1988; T. Imagawa, J. S. Smith, R. Coronado, and K. P. Campbell. J. Biol. Chem. 262: 10636 10643, 1987; L. Hymel, M. Inui, S. Fleischer, and H. Schindler, Proc. Natl. Acad. Sci. USA 85:441-445, 1988) was paralleled by an increased binding site density (Bmax) for ryanodine binding in denervated muscle TC. The effects of ruthenium red, a Ca2+ release blocker, on Ca2+ loading rate and Ca2+-ATPase activity suggested that TC from denervated muscle were less permeable to Ca2+. After active Ca2+ loading, both doxorubicin and caffeine induced Ca2+ release from isolated TC, yet Ca2+ release rates were reduced in denervated muscle TC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551175 TI - Increased ACTH response to corticotropin-releasing factor in cold-adapted rats in vivo. AB - We have recently reported that chronically repeated restraint stress results in improved cold tolerance in rats via an increased activity of nonshivering thermogenesis, a characteristic metabolic change observed during cold adaptation, suggesting the presence of cross-adaptation between cold and stress. It is well established that the hypothalamic-pituitary-adrenal (HPA) axis is activated in various stress responses. In the present study, therefore, we examined whether cold adaptation would alter the adrenocorticotropic hormone (ACTH)-releasing state in vivo using freely moving, conscious rats chronically implanted with intra-atrial cannulas. There was no difference in the basal levels of plasma ACTH between warm control and cold-adapted rats. On the other hand, the ACTH response to the intravenous administration of corticotropin-releasing factor (CRF; 2 micrograms/animal) was significantly elevated in cold-adapted rats. However, the injection of 10 micrograms of CRF, which was considered as a dose to elicit the maximal ACTH response, resulted in similar ACTH release patterns between the two groups. These changes in the responsiveness of ACTH secretion have been observed in rats chronically exposed to stressful conditions. The results demonstrated in the present study, therefore, provide further evidence for our hypothesis that there may exist cross-adaptation between cold and nonthermal stress. PMID- 2551174 TI - Identification of a 185-kDa band 3-related polypeptide in oxyntic cells. AB - Polyclonal antibodies to the purified mouse erythrocyte anion exchange protein (band 3) and to a conserved COOH-terminal peptide of mouse band 3 (alpha-Ct) recognized a single major 185-kDa polypeptide in immunoblots of a membrane fraction prepared from rabbit gastric glands. Competition studies revealed that the epitopes shared between the rabbit gastric 185-kDa antigen and the approximately 100-kDa mouse erythrocyte band 3 protein are restricted to the COOH terminal domain of band 3, which is known to contain the catalytic site for anion exchange activity. Immunofluorescence microscopy was used to demonstrate that this band 3-related polypeptide is associated with the plasma membrane in a subpopulation of gastric gland cells composed exclusively of oxyntic cells, as judged by the coincidence of immunofluorescence with alpha-Ct and with a monoclonal antibody to the gastric H+-K+-ATPase. This alpha-Ct-reactive antigen was further localized to the cytoplasmic face of the basolateral membrane of oxyntic cells, which correlates well with the physiologically determined site of anion exchange activity. These data demonstrate the presence in gastric oxyntic cells of a novel member of the family of proteins related to the erythrocyte anion exchanger. The possibility that the 185-kDa polypeptide is an anion exchanger is discussed. PMID- 2551176 TI - Beta-endorphin and islet hormone release in humans: evidence for interference with cAMP. AB - The present studies were undertaken to characterize further the influence of synthetic human beta-endorphin (0.5 mg/h) on insulin and glucagon responses to intravenous glucose in humans. Infusion of beta-endorphin in 10 normal volunteers caused a clear-cut inhibition of the overall insulin responses to a glucose pulse (0.33 g/kg iv) with values of glucose disappearance rates in the diabetic range [0.89 +/- 0.09 (P less than 0.01) vs. saline 1.82 +/- 0.15%/min]. Glucose-induced glucagon suppression was significantly lower during beta-endorphin, a fact that could have contributed to the reduced glucose utilization rates. The infusion of theophylline (150 mg + 350 mg/h) to increase the intracellular cAMP activity by inhibiting phosphodiesterase completely reversed the inhibitory effect of beta endorphin on glucose-induced insulin secretion. As a consequence, glucose disappearance rates rose to 1.77 +/- 0.18%/min. Theophylline did not influence significantly the glucagon-releasing effect of beta-endorphin as well as the reduced glucagon suppression. An infusion of exogenous calcium (100 mg as iv bolus + 5 mg/min) to raise serum calcium in the hypercalcemic range (15 mg/dl) and lysine acetylsalicylate (72 mg/min) to block the synthesis of endogenous prostaglandin E did not interfere with the inhibiting effect of beta-endorphin on insulin secretion. These data confirm that beta-endorphin stimulates glucagon and inhibits basal and glucose-stimulated insulin secretion and suggest that the opioid influences the intraislet adenylate cyclase activity. PMID- 2551177 TI - Effects of ketamine and fentanyl on lung metabolism in perfused rat lungs. AB - The effects of ketamine and fentanyl on serotonin (5-hydroxytryptamine; 5-HT) metabolism, angiotensin-converting enzyme (ACE), and protein synthesis (PS) were investigated in an isolated lung model. Rat lungs were perfused in situ with a blood-free physiological salt solution. The pulmonary vasculature was exposed to ketamine (0.005-2.1 mM) or fentanyl (1.8-4.5 microM) for up to 2 h. After 1 h, accumulation of 5-[14C]hydroxyindoleacetic acid (5-HIAA) by the lung was monitored as an index of 5-HT metabolism. ACE activity was estimated from hydrolysis of [3H]benzoylphenylalanyl-alanyl-proline, a synthetic substrate for the enzyme. [3H]phenylalanine was added to the perfusate after 1 h, and its incorporation into acid-precipitable lung protein was measured over the subsequent hour. Ketamine inhibited 5-HT uptake in a concentration-related manner. The inhibition was characterized as competitive and reversible. Fentanyl had no effect on lung 5-HIAA accumulation. Neither drug altered ACE activity or protein synthesis over the concentration ranges tested. The results indicate an action by ketamine that inhibits the 5-HT membrane-transport process. The different effects observed by ketamine and fentanyl on this process could contribute to the diverse pharmacological properties of these two drugs. PMID- 2551178 TI - Recovery of 13C in breath from NaH13CO3 infused by gut and vein: effect of feeding. AB - Estimates of substrate oxidation obtained from appearance of 13C or 14C from tracers in breath must be corrected for retention of labeled carbon in the body. We aimed to determine the effect of a defined experimental diet and metabolic status on recovery of infused Na [13C]bicarbonate in breath. Six healthy male subjects consumed an experimental diet for 7 days before receiving a continuous infusion of formula without tracer on day 8 and received either an intragastric (ig) or intravenous (iv) infusion of Na [13C]bicarbonate on day 9 or 11 during a 4-h postabsorptive (PA), 4-h continuously fed period. A trend toward increasing PA breath enrichment during the first 7 diet days approached statistical significance (P = 0.051), whereas breath enrichments measured 3 h postbreakfast were consistently higher than PA values throughout and did not change over the 7 day period. Breath enrichments during a 4-h continuous ig infusion of formula without tracer on day 8 rose 2.0 +/- 0.5 atom percent excess (APE).10(-3) above base line (P less than 0.001, ANOVA). In the tracer studies, breath enrichments were similar for the ig and iv routes of tracer infusion. For the ig infusion the fraction of infused Na [13C]bicarbonate recovered in breath as 13CO2 was 0.74 +/- 0.02 for the PA period and 0.79 +/- 0.02 for the fed period. For the iv infusion the fraction recovered was 0.70 +/- 0.04 for the PA period and 0.82 +/- 0.03 for the fed period. Fractional recoveries were not significantly different for ig and iv routes of administration but were different for PA and fed periods (P less than 0.0001, 2-way ANOVA). The fractional recoveries for the fed period obtained here were similar to the value 0.81 reported in a number of other studies. Recovery of tracer in breath increased linearly with O2 uptake and CO2 production, suggesting that factors affecting respiratory gas exchange may alter recovery. We conclude that the primary factor determining label recovery is the immediate and recent nutritional status of the host. PMID- 2551179 TI - pH regulation in hepatoma cells: roles for Na-H exchange, Cl-HCO3 exchange, and Na-HCO3 cotransport. AB - Regulation of intracellular pH (pHi) was studied in Fu5, a rat hepatoma cell line that maintains a variety of differentiated functions. Microspectrofluorimetry of the pH-sensitive dye 2',7'-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF) was used to measure pHi in 10-15 cells growing on cover glasses that were mounted in a flow-through chamber on the stage of a microscope. In N-2 hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES)-buffered solutions, pHi was 7.14, and intrinsic buffer capacity was inversely related to pHi. Amiloride (0.1 mM) caused pHi to decrease by 0.33 pH units in 4 min. Recovery from an acid load (using either NH4 prepulse technique or Na-free solutions) was completely blocked by amiloride. In HCO3-CO2-buffered solutions, pHi was 7.15, and buffer capacity was relatively insensitive to pHi between pHi of 6.6 and 7.2. Amiloride caused pHi to decrease by only 0.09 units. Recovery from an acid load was Na dependent, occurred in Cl-free solutions, and was totally blocked by the combination of amiloride plus 0.5 mM dihydro-4,4'-diisothiocyanostilbene-2,2' disulfonic acid (H2DIDS); recovery occurred when either amiloride or H2DIDS was removed. Removal of external Cl caused a rapid, H2DIDS-blockable alkalinization that was faster in HCO3-CO2 than in HEPES. The apparent Km for Clout for relaxation of Cl-free alkalinization was 4.5 mM. Rate of HCO3 transport during Cl free treatment increased at alkaline resting pHi. It is concluded that Fu5 cells have two Na-dependent base-loading mechanisms and an acid-loading Cl-HCO3 exchanger. In solutions containing HCO3-CO2, the Na-H exchanger accounts for approximately 40% of recovery from an acid load, and a Na-HCO3 cotransporter accounts for the remainder. Recovery from an alkaline load appears to occur through the activity of the Cl-HCO3 exchanger. PMID- 2551180 TI - Volume-regulatory K+ fluxes in the isolated perfused rat liver: characterization by ion transport inhibitors. AB - Net hepatic release and uptake of K+ were examined in isolated perfused rat livers subjected to a 10-min period of hypotonic stress. Effluent Na+, K+, and Ca2+ activities were monitored throughout. Initiation and termination of hypotonic stress triggered sharp transient (less than 1 min) changes in effluent ion activities that indicated net water movement into and out of the liver, respectively. In addition, hypotonic stress caused a large transient net release of hepatic K+, whereas return to isotonicity triggered a transient net hepatic K+ uptake. The hypotonically induced K+ release was inhibited by 2 mM barium (95%) and by 1 mM quinine (60%). Net K+ influx, on the other hand, was inhibited by 1 mM ouabain (100%) and by 1 mM amiloride (50%). Osmotically induced K+ fluxes were not significantly affected by bicarbonate removal and were only partially inhibited by 0.1 mM 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) or bumetanide. The results suggest that K+ conductance increases during hypotonic stress, whereas return to isotonicity induces a ouabain-sensitive K+ uptake partly because of increased Na+-H+ exchange. These mechanisms probably participate in regulatory volume decrease and regulatory volume increase, respectively. PMID- 2551181 TI - Ursodeoxycholate-induced changes in hepatic Na+-H+ exchange and biliary HCO3- excretion. AB - Ursodeoxycholate (UDC)-induced HCO3- -rich choleresis may be due to activation of sinusoidal Na+-H+ exchange followed by an increase in intracellular pH (pHi) and HCO3- excretion via canalicular Cl- -HCO3- exchange. To test this hypothesis, we studied the effect of UDC and tauroursodeoxycholate (TUDC) on net H+ efflux from perfused rat livers and pHi in isolated hepatocytes in the presence and absence of amiloride. UDC-induced increases in biliary HCO3- concentration and excretion were inhibited by amiloride. However, these increases were temporally associated with an initial decline in H+ efflux and pHi followed by a gradual recovery toward base line. The initial decline in H+ efflux was associated with a rapid uptake of UDC. Amiloride inhibited only the recovery phases of H+ efflux and pHi. TUDC increased amiloride-sensitive H+ efflux without affecting biliary [HCO3-] and decreased pHi in the presence but not in the absence of amiloride. Amiloride decreased TUDC-induced choleresis and HCO3- excretion most likely by decreasing TUDC excretion. TUDC decreased biliary [Cl-] and increased hepatic O2 uptake more than UDC. We conclude that a rapid influx of UDC in the protonated form decreases pHi and net H+ efflux initially. The recovery phase is due to Na+-H+ exchange activated by decreased pHi and possibly by UDC and increased cellular respiration. TUDC indirectly stimulates Na+-H+ exchange most likely by increasing cellular respiration. UDC-induced HCO3- -rich choleresis, which is observed at a time when both net H+ efflux and pHi are less than control values, is unlikely to be due to a direct activation of Na+-H+ exchange. PMID- 2551182 TI - Signal transmission after GLP-1(7-36)amide binding in RINm5F cells. AB - Glucagon-like peptide-1(7-36)amide [GLP-1(7-36)amide], probably representing an important incretin, binds to receptors on RINm5F cells resulting in an adenosine 3',5'-cyclic monophosphate increase. Guanine nucleotides (GTP, GTP-gamma-S, GDP beta-S) decreased the binding of GLP-1(7-36)amide to receptors on RINm5F cell membranes. Further analysis revealed that GTP (10(-4) M) decreased the receptor affinity with an increase of the Kd from 2.5 +/- 0.99 x 10(-10) M to 9.43 +/- 2.16 x 10(-10) M. In cross-linking experiments the amount of labeled peptide linked to receptors was reduced in the presence of GTP (10(-4) M). Further studies investigated the involvement of membrane depolarization or changes in the cytosolic free calcium level in the intracellular signaling of GLP-1(7-36)amide induced insulin secretion. In contrast to fuel and nonfuel secretagogues, GLP-1(7 36)amide did not cause a depolarization of the membrane potential. This was unaffected by various glucose concentrations (0-20 mM) or by previous cell depolarization by D-glyceraldehyde. Similarly, the cytosolic calcium concentration remained unchanged after addition of GLP-1(7-36)amide (10(-12)-10( 8) M). The effect of guanine nucleotides on binding of GLP-1(7-36)amide indicates that the action of the peptide is mediated by the adenylate cyclase system. GLP 1(7-36)amide binding neither changed the membrane potential nor altered the intracellular calcium concentration, making an involvement of the inositol 1,4,5 trisphosphate pathway or an activation of protein kinase C in the postreceptor signaling after GLP-1(7-36)amide binding unlikely. PMID- 2551183 TI - Carbachol-induced down-regulation of high-affinity receptors for vasoactive intestinal peptide. AB - When dispersed acini from guinea pig pancreas are first incubated with carbachol, the subsequent binding of 125I-vasoactive intestinal peptide (VIP) is inhibited during a second incubation. This inhibitory action of carbachol on binding of 125I-VIP depends on time, temperature, and the concentration of carbachol in the first incubation and can be blocked by atropine. First incubating acini with A23187, ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), cholecystokinin octapeptide, bombesin, or 12-O-tetradecanoylphorbol-13 acetate does not alter binding of 125I-VIP. Adding EGTA to the first incubation medium abolishes the effect of carbachol on binding of 125I-VIP. In control acini or acini first incubated with carbachol, approximately half of the bound 125I-VIP can be stripped by acetic acid. 125I-VIP interacts with two distinct classes of receptors on pancreatic acini. One has a high affinity for VIP (Kd, 1 nM); the other has a low affinity for VIP (Kd, 2 microM). First incubating acini with carbachol decreases the number but not the affinity of high-affinity VIP receptors with no change in the number or affinity of low-affinity VIP receptors. Pancreatic acini possess two classes of muscarinic cholinergic receptors: one has a high affinity (Kd, 4 microM) and the other has a low affinity (Kd, 698 microM) for carbachol. The dose-response curve for carbachol-induced inhibition of binding of 125I-VIP and that for occupation of low-affinity muscarinic cholinergic receptors by carbachol are similar.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551184 TI - Radical production during in vivo intestinal ischemia and reperfusion in the cat. AB - Free radical formation was studied with electron spin resonance during 2 h of intestinal ischemia in the cat, at a blood flow less than 5 ml.min-1.100 g-1, followed by 30-min reperfusion. A modification of the spin trapping technique was used to stabilize highly reactive free radicals. The rate of secondary radical formation was 0.32 +/- 0.06 mumol.min-1.100 g intestine-1 before ischemia and increased to a maximum of 0.66 +/- 0.09 mumol.min-1.100 g-1 during the first minutes of reperfusion (mean +/- SE, n = 5). This could be prevented either by maintaining intestinal blood flow at 8-15 ml.min-1.100 g-1, by administering allopurinol before and during ischemia, or by perfusing the intestinal lumen with an O2-saturated buffer solution during ischemia, resulting in maximum rates of radical production during reperfusion of 0.37 +/- 0.04 (n = 6), 0.33 +/- 0.04 (n = 5), and 0.39 +/- 0.13 mumol.min-1.100 g-1 (n = 5), respectively. The results demonstrate that free radicals are produced in the intestine during reperfusion after a period of reduced blood flow below a certain critical level, and that inhibition of xanthine oxidase and prevention of hypoxia will eliminate this radical production. PMID- 2551185 TI - Mitogenic signals for thrombin in mesangial cells: regulation of phospholipase C and PDGF genes. AB - Thrombin is a proteolytic enzyme of diverse biological activities, which is produced during activation of the coagulation pathway. In addition, thrombin is a mitogen for fibroblasts and endothelial cells. Intraglomerular thrombosis and cell proliferation are common pathological features of several glomerular diseases. We studied the effect of thrombin on deoxyribonucleic acid (DNA) synthesis in cultured human mesangial cells and explored mechanisms of signal transduction involved. Bovine and human thrombin caused dose-dependent increases in DNA synthesis, inositol trisphosphate, and cytosolic calcium [(Ca2+)i]. A threefold increase in inositol-3-trisphosphate (IP3) levels was observed as early as 10 s after the addition of thrombin, whereas increases in (Ca2+)i occurred within 5-10 s and declined rapidly. Stimulation of mesangial cells by thrombin resulted in induction of messenger ribonucleic acids (mRNAs) encoding platelet derived growth factor (PDGF) A- and B-chains. This was associated with an enhanced secretion of PDGF-like protein. These data provide mechanisms by which thrombin may regulate mesangial cell function in disease states. PMID- 2551186 TI - Metabolic alterations in proximal tubule suspensions obtained from ischemic kidneys. AB - A viable suspension of proximal tubules that had sustained an in vivo ischemic injury was harvested, and cellular integrity and viability were determined. The histopathological appearance of this preparation has characteristic features of an ischemic injury and ATP levels were comparable to those observed with nuclear magnetic resonance spectroscopy in vivo. Sprague-Dawley rats were subjected to 45 min of bilateral renal artery ischemia and the kidneys were allowed to reperfuse for either 15 min, 2 h, or 24 h before the harvest of the proximal tubule suspension. There was a decrease in base-line oxygen consumption from 34 +/- 0.8 nmol O2.min-1.mg protein-1 to 22 +/- 0.6 at 15 min of reflow. This decline in oxygen consumption persisted during the first 2 h of reflow and returned to control levels by 24 h. Residual respiration in the presence of ouabain was similar at all reflow intervals suggesting that the decrease in basal O2 consumption was related to decreased Na+-K+-ATPase in situ. In contrast, there was no significant difference in Na+-K+-ATPase activity when determined chemically under Vmax conditions in all experimental groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551187 TI - Prostaglandin E2 inhibits Na+-K+-ATPase activity in the inner medullary collecting duct. AB - Prostaglandin E2 (PGE2) is natriuretic and inhibits collecting duct sodium transport by poorly defined mechanisms. To determine the mechanism of this inhibition, we have studied the effect of PGE2 on ouabain-sensitive (transport dependent) oxygen consumption (QO2), ouabain-sensitive 86Rb+ uptake and ouabain sensitive ATPase activity in fresh suspensions of rabbit inner medullary collecting duct cells, as well as Na+-K+-ATPase activity in inner medullary membranes. PGE2 (10(-5) M) reduced total QO2 by 21.6 +/- 2.3% (mean +/- SE) and reduced the ouabain-sensitive component of QO2 in IMCD cells. PGE2 failed to inhibit QO2 in the absence of sodium or in the presence of ouabain and blunted the increase in QO2 in response to amphotericin B. These results suggested that PGE2 inhibited Na+-K+-ATPase activity. Inhibition of pump activity was confirmed by measurements of 86Rb+ uptake: PGE2 (10(-5) M) reduced ouabain-sensitive 86Rb+ uptake by 57% at 10 s without altering equilibrium uptake. Furthermore, PGE2 (10( 6) M) reduced ouabain-sensitive ATPase activity by 46% in permeabilized inner medullary collecting duct cells. PGF2 alpha (10(-5) M) did not significantly alter QO2, 86Rb+ uptake, or Na+-K+-ATPase activity. These results demonstrate that PGE2 inhibits inner medullary collecting duct Na+-K+-ATPase activity and suggest a role for this inhibition in the natriuretic effect of PGE2. PMID- 2551188 TI - Studies of the mitogenic effect of serotonin in rat renal mesangial cells. AB - A vasoactive inflammatory amine, serotonin, stimulates DNA synthesis in rat glomerular mesangial cells in a dose-dependent manner and acts synergistically with either insulin or epidermal growth factor (EGF). The combined effects of 10( 6) M serotonin and these peptide hormones are nearly equal to those induced by 10% fetal bovine serum. Serotonin stimulates the turnover of polyphosphoinositols resulting in a transient rise in intracellular free Ca2+ concentration, as measured either with the photoprotein aequorin, or with fura-2. This is accompanied by a transient increase in 45Ca2+ efflux from prelabeled cells. Serotonin also induces a prompt and sustained threefold increase in Ca2+ influx rate across the plasma membrane and a rapid and sustained twofold increase in cellular 1,2-diacylglycerol content. In addition, there is an increase in the extent of phosphorylation of an acidic 80-kDa protein, a putative substrate for protein kinase C. Activators of protein kinase C (including phorbol 12-myristate 13-acetate or 1,2-dioctanoylglycerol) mimic the mitogenic effect of serotonin. The effect of serotonin on cell proliferation is partially inhibited in a reversible manner by LiCl. Treatment of mesangial cells with insulin plus EGF for 60 min leads to a small but consistent increase in the content of inositol phosphates and 1,2-diacylglycerol. Their effects are additive to those of serotonin. Moreover, insulin and EGF significantly stimulate the phosphorylation of the 80-kDa protein, and potentiate the serotonin-induced phosphorylation of this protein.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551189 TI - Refined membrane preparations mask ischemic fall in myocardial beta-receptor density. AB - Most of the previous studies of ischemic myocardial beta-adrenergic receptors have employed membrane preparations in which the initial pellet from the myocardial homogenate spun at a low speed was discarded. We studied changes in beta-adrenergic receptor density ([125I]-iodocyanopindolol; Bmax) during 30 min of coronary occlusion in surgically anesthetized open-chest rabbits using just such a pellet [homogenized heart spun at 1,000 g (1,000-g pellet)], as well as a second pellet from the supernatant of the first pellet [spun at 40,000 g (40,000 g pellet)]. Bmax fell during acute ischemia in the 1,000-g pellet [46.8 +/- 6.1 vs. 21.6 +/- 2.4 (SE) fmol/mg protein; P less than 0.01; n = 7] but did not change in the 40,000-g pellet [46.8 +/- 6.5 vs. 47.9 +/- 2.6 (SE) fmol/mg protein; P = NS; n = 6]. The 1,000-g pellet contained 70.0 +/- 8.1% of the beta adrenergic receptors measured between the two preparations (P less than 0.05; n = 8) and all of the histamine H2-receptors; therefore, to minimize receptor loss and other potential artifacts, unspun myocardial homogenate was studied. An ischemic decrease in Bmax was still observed [32.9 +/- 2.0 vs. 20.9 +/- 4.1 (SE) fmol/mg protein; P less than 0.05; n = 5]. These results support the use of data from cruder myocardial membrane preparations (e.g., 1,000-g pellet or unspun homogenate), which may be of greater pathophysiological relevance than data derived from a standard more-refined preparation (40,000-g pellet). PMID- 2551190 TI - Mechanism for depression of heart sarcolemmal Ca2+ pump by oxygen free radicals. AB - To understand the involvement of changes in sulfhydryl groups in causing depression of the sarcolemmal Ca2+-pump activities, this study was undertaken to examine the effects of oxygen free radicals on rat heart sarcolemmal sulfhydryl groups, Ca2+-stimulated adenosinetriphosphatase (ATPase), and ATP-dependent Ca2+ accumulation. In addition, the effects of sulfhydryl reagents such as dithiothreitol, cysteine, and N-ethylmaleimide on Ca2+-pump activities were investigated. The inhibition of sarcolemmal Ca2+-pump activities by O2-. (xanthine + xanthine oxidase) and H2O2 was decreased by the addition of dithiothreitol or cysteine in a dose-dependent manner. N-ethylmaleimide also showed inhibitory effects on Ca2+-pump activities both in a dose- and time dependent manner; dithiothreitol and cysteine prevented changes in Ca2+-pump activities because of N-ethylmaleimide. Heart sarcolemmal sulfhydryl groups were depressed by O2-., H2O2, and .OH (H2O2 + Fe2+) both in a dose- and time-dependent manner. Superoxide dismutase, catalase, and D-mannitol showed protective effects on the sulfhydryl group depression by O2-., H2O2, and .OH, respectively. A significant correlation between changes in sarcolemmal Ca2+-stimulated ATPase activity and sarcolemmal sulfhydryl groups was seen. These results indicate that oxygen free radicals may depress the heart sarcolemmal Ca2+-pump activities by modifying the sulfhydryl groups in the sarcolemmal membrane. PMID- 2551191 TI - Effect of transit time on metabolism of a pulmonary endothelial enzyme substrate. AB - Fractional hydrolysis (M) of the synthetic angiotensin-converting enzyme (ACE) substrate [3H]benzoyl-Phe-Ala-Pro (BPAP) on passage through the isolated dog lung lobe was found to be relatively independent of flow rate and transit time (t). The most commonly expressed explanation for this kind of observation is that recruitment of ACE-containing surface area occurs when flow is increased. To test this, as well as other hypotheses that might explain the behavior of this substrate, we compared M obtained after the first pass of a BPAP-containing bolus through isolated rabbit lungs with that obtained after two sequential passes through the lungs. In this way, we could double t with no change in flow or vascular pressure. We found that, when the second pass occurred within a few seconds of the first, M after both the first and second pass was only slightly larger than that after the first pass alone. If the time between passes was increased to a few minutes, M after the second pass was substantially increased. These results are contrary to the recruitment hypothesis and suggest that this substrate may exist in alternative forms that are in slow equilibrium relative to the capillary t. When albumin was present in the perfusate, an albumin-bound fraction appeared to be one such alternative form. However, experiments carried out using protein-free perfusate suggest the possibility that conformational variants of the substrate may also exist. PMID- 2551192 TI - Tension and electrolyte changes with Na+-K+ pump inhibition in rat papillary muscle. AB - Myocardial ischemic injury results in altered membrane integrity, energy depletion, and electrolyte shifts leading to accumulation of intracellular Ca. However, analysis of the direct effects of Ca accumulation is complicated by other concomitant cellular changes produced by ischemia. The purpose of this study was to examine the effects of Ca loading in rat papillary muscles produced by Na+-K+ pump inhibition in oxygenated K+-free buffer. Changes in contractile characteristics, high energy phosphate, and elemental concentrations of subcellular compartments were measured. Electron probe X-ray microanalysis was used to assess elemental concentrations in cryosections. After 3 h of Na+-K+ pump inhibition, resting tension (RT) increased to 164% and developed tension (DT) fell to 16.8% of control values. One hour after return to complete buffer, RT and DT partially recovered but remained significantly different from the 180 to 240 min values for the control muscles. Electron probe X-ray microanalysis showed increases in cytoplasmic and mitochondrial Na and Ca and a decrease in K during Na+-K+ pump inhibition. Mitochondrial Ca was greater than 100-fold greater than Ca in control mitochondria. Morphologically, the majority of cells showed ultrastructural damage. The mean ATP level was 20% of control. After 1 h of recovery, the cells appeared more heterogeneous, and the mean mitochondrial Ca decreased, whereas mean cytoplasmic Ca increased. Further statistical analysis showed a bimodal distribution for Na, Ca, K, Mg, and Cl, which coincided with the morphologically mixed population of cells. This suggests that replacement of extracellular K+ was associated with restored electrolyte gradients in some cells and the persistent or further alteration of electrolytes in others. These results suggest that variable Ca accumulation and associated ATP depletion without the compounding effects of ischemia lead to cell injury similar to reperfusion injury reported in ischemic myocardium. PMID- 2551193 TI - Kinetic analysis of electrogenic 2 Na+-1 H+ antiport in crustacean hepatopancreas. AB - Na+ uptake by short-circuited brush-border membrane vesicles of the hepatopancreatic epithelium from the freshwater prawn Macrobrachium rosenbergii was Cl- independent, amiloride sensitive, and stimulated by a transmembrane proton gradient ([H+]i greater than [H+]o). Na+ influx (3-s uptake) was a sigmoidal function of [Na]o (2.5-150 mM), when pHi = 6.0, pHo = 8.0, and followed the Hill equation for binding cooperativity [maximal Na+ influx (Jm) = 140.6 nmol mg-1s-1; affinity constant (K') = 82.2 mM Na+; Hill coefficient (n) = 2.07]. Influx kinetic analyses at physiological conditions suggested two external cation binding sites shared by Na+ and H+ (proton dissociation constant Pk1 = 5.7; Pk2 = 4.0) and a single internal cation site used only by H+ (Pk = 6.5). Amiloride was a competitive inhibitor of Na+ transport at both external binding sites (Ki1 = 50 microM; Ki2 = 1,520 microM). Electrogenic Na+-H+ exchange by these vesicles was demonstrated using an equilibrium-shift method of analysis and a transmembrane electrical potential difference as the only driving force for transport. In addition, electrogenic net Na+ influx (3-s uptake) was observed in vesicles loaded with 5 mM 22Na at pH 7.0 and exposed to media containing several 22Na or proton concentrations. Results suggest the following exchange model: low [Na]o, (1 Na+ and 1 H+)-1 H+; high [Na+]o, 2 Na+-1 H+. This antiport mechanism may account for two major functional operations of the gastrointestinal tract in these animals: 1) proton secretion against considerable concentration gradients leading to stomach luminal acidification, and 2) Na+ absorption from lumen to cytoplasm potentially making a significant contribution to organismic ion balance. PMID- 2551194 TI - Human selenite metabolism: a kinetic model. AB - A model is developed to describe the kinetics of sodium selenite metabolism in humans, based on plasma, urine, and fecal samples obtained from six subjects over a 4-wk period after a single oral 200-micrograms dose of the enriched stable isotope tracer 74Se. The model describes absorption, distributed along the gastrointestinal tract, and enterohepatic recirculation. The model includes four kinetically distinct plasma components, a subsystem consisting of the liver and pancreas, and a slowly turning-over tissue pool. For the six subjects, the ranges of mean residence times for the four plasma components are, respectively, 0.2-1.1 h, 3-8 h, 9-42 h, and 200-285 h; for the hepatopancreatic subsystem 4-41 days; and for the tissue pool 115-285 days. Approximately 84% of the administered dose was absorbed, and after 12 days approximately 65% remained in the body. The model predicts that after 90 days approximately 35% of this Se would be retained, primarily in the tissues. Separating Se metabolism into several distinct kinetic components is a first step in identifying the efficacious, nutritious, and toxic forms of the element. PMID- 2551195 TI - Prior immobilization stress alters adrenal hormone responses to hemorrhage in rats. AB - The effect of prior immobilization stress (IMO) on the plasma epinephrine (EPI), norepinephrine (NE), adrenocorticotropic hormone (ACTH), and corticosterone (CS) responses to acute hemorrhage was studied in conscious male rats with chronic catheters in tail artery, using two combinations of IMO and hemorrhage. IMO per se led to significant increases of EPI, NE, ACTH, and CS in all animals. Hemorrhage of 25% of estimated blood volume (EBV) performed immediately after 150 min IMO caused exaggerated release of EPI and NE, whereas CS remained unchanged at the level previously elevated by IMO. ACTH response to initial blood loss of 12.5% was diminished in previously immobilized rats. Hemorrhage of 35% EBV after 60 min IMO and a 10-min recovery period also resulted in potentiated increases in EPI and NE, suppressed ACTH secretion, and no further change in stress-elevated CS concentration. No differences between groups were observed in relative mean arterial blood pressure, plasma protein, and lactate responses to 35% hemorrhage. Posthemorrhagic increase of plasma concentration of several tissue enzyme activities was significantly higher in prestressed rats. Furthermore, 24-h mortality rate increased by 49%. In summary, our results indicate that prior IMO potentiated activation of the sympathoadrenomedullary system and suppressed ACTH response to subsequent hemorrhage. Altered neuroendocrine responsiveness and stress-induced prehemorrhagic tissue damage may play roles in the increased susceptibility of the organism to blood loss. PMID- 2551197 TI - The effect of fentanyl on basal and stimulated plasma levels of atrial natriuretic peptide. PMID- 2551196 TI - Determinants of polychlorinated biphenyls (PCBs), polybrominated biphenyls (PBBs), and dichlorodiphenyl trichloroethane (DDT) levels in the sera of young children. AB - Serum samples from 285 4-year-old Michigan children were evaluated for levels of 11 environmental contaminants. Polychlorinated biphenyls (PCBs) were found in half the samples tested; polybrominated biphenyls (PBBs) in 13-21 percent; dichlorodiphenyl trichloroethane (DDT), in more than 70 percent. Nursing (Mothers' milk) was the principal source of these exposures. Congener-specific analysis documented the presence of at least one highly toxic PCB congener, 2,3',4,4',5-pentachlorobiphenyl. The data demonstrate the multigenerational impact of female exposure to persistent organic environmental contaminants. PMID- 2551198 TI - Release of immunoreactive arachidonate metabolites by equine endometrium in vitro. AB - The ability of equine endometrium to release prostaglandin (PG) F, PGE2, and leukotriene (LT) B4 was studied in vitro, using endometrial tissue from diestrous mares. Because of the high cross-reactivity of the PGF antiserum with PGF1 alpha and with PGF2 alpha, results were quoted as total immunoreactive PGF. Significant concentrations of these arachidonate metabolites were released into tissue culture medium between 1 and 24 hours of incubation. Significantly higher concentrations of PGE, but not of PGE2 or LTB4, were released from endometria of mares with chronic endometritis than from genitally normal mares. Prostaglandin F was released only in low concentrations from the endometrium of a mare with pyometra, but concentrations of PGE2 and LTB4 were similar to those of genitally normal mares. PMID- 2551199 TI - Antibody response of pseudorabies virus subunit-vaccinated pigs to viral nucleocapsid proteins following low-dose virus challenge of immunity. AB - The antibody response to pseudorabies virus nucleocapsid proteins (NCP) was evaluated by the western immunoblot analysis before and after challenge of immunity by nasal inoculation of 10(2.3) plaque-forming units of virus in 10 pigs that had been vaccinated with pseudorabies virus envelope glycoproteins. Antibody to 5 NCP with molecular mass of 140, 63, 41, 34, and 23 kD was first detected in vaccinated and nonvaccinated pigs on day 14 after challenge of immunity. Antibody to 2 of the 5 NCP continued to be detected through day 113 in 9 of 10 vaccinated pigs. Beyond day 32, antibody to NCP was not detected in 1 vaccinated pig. The 23 , 34-, and 41-kD proteins were the most immunogenic. Antibody to each of these proteins was first detected on day 14 in 10, 10, and 8 pigs, respectively. Seven, 6, and 8 pigs, respectively, were antibody-positive for these proteins on day 113. The 140- and 63-kD proteins were the least immunogenic. Antibody to these proteins was detected in 8 and 9 pigs, respectively, on day 14, and in 4 and 5 pigs, respectively, on day 113. Chi-square analysis for dependency indicated that the antibody response to the 140- and 63-kD proteins was interdependent. These results suggested that combinations of NCP may be useful as nonvaccine diagnostic antigens. PMID- 2551201 TI - Isolation and trypsin-enhanced propagation of turkey enteric (bluecomb) coronaviruses in a continuous human rectal adenocarcinoma cell line. AB - Turkey enteric coronavirus (TCV) from intestinal contents of diarrheal poults was isolated and serially propagated in HRT-18 cells, an established cell line derived from a human rectal adenocarcinoma. In these cells, TCV induced cytopathic changes, including polykaryocytosis, which depended on trypsin in the medium and incubation at 41 C. Viral antigens could be demonstrated in the cytoplasm by immunofluorescence, and extracellular virus was detected by an ELISA and negative electron microscopy. The cell-free virus had characteristics of TCV: shape, surface projections, buoyant density of 1.18 to 1.20 g/ml in sucrose, and hemagglutination of rat RBC. The one-step growth curve was complete by postinoculation hours 14 to 16, and maximal titers reached 9 to 9.5 log10 TCID50/ml during 5 passages, after which the titer remained stable. Electron microscopic examination of infected cell monolayers revealed budding of typical coronavirus particles through intracytoplasmic membranes and accumulation of complete virus in cytoplasmic vesicles. Late in the infection, aggregated progeny vial particles were detected near the outer surface of infected cells. One-day old turkey poults inoculated orally with tissue culture-adapted TCV isolates developed mild to severe diarrhea. PMID- 2551200 TI - Infection of the middle nasal meatus of calves with Pasteurella haemolytica serotype 1. AB - Eight healthy nonstressed calves were inoculated with Pasteurella haemolytica serotype 1, by instilling a broth culture into the middle nasal meatus of the left nostril. The inoculated left nostrils shed P haemolytica from the ventral nasal meatus at a steady rate for a mean of 7 days, whereas the uninoculated right nostrils of the same calves shed P haemolytica sporadically and in lower concentrations. The duration, frequency, and concentration of P haemolytica shed from the inoculated nostrils was significantly (P less than 0.05) greater than from the nostrils of other healthy calves that had been exposed by instilling the culture into the ventral nasal meatus of both nostrils in a previous study. The concentration of antibodies (IgG, IgA, and IgM) to P haemolytica increased significantly (P less than 0.05) in serum and nasal secretions after exposure. Four weeks after initial P haemolytica exposure, calves were exposed to infectious bovine rhinotracheitis virus and became clinically ill. Four calves were induced to shed P haemolytica from both nostrils by the virus infection; thus, they were harboring the bacterium and were susceptible to active recolonization. Four calves were not induced to shed P haemolytica. The apparent reason was not that they were resistant to active colonization, but that they were no longer harboring the bacterium, because they became active shedders after they were reinfected with P haemolytica. PMID- 2551202 TI - Furosemide and sodium bicarbonate-induced alkalosis in the horse and response to oral KCl or NaCl therapy. AB - Metabolic alkalosis was induced in 10 clinically normal horses by administration of furosemide (1 mg/kg of body weight, IM) followed 4.5 hours later by sodium bicarbonate (NaHCO3; 500 g in 8 L water) via nasogastric tube. Furosemide diuresis resulted in a mean weight loss of 21.1 kg, which was associated with small, but significant, increases in venous blood pH, bicarbonate, and plasma protein concentrations (P less than 0.001), while plasma potassium, chloride, and calcium concentrations declined significantly (P less than 0.001). Oral administration of the hypertonic NaHCO3 solution resulted in clinical evidence of hypovolemia, which was accompanied by a marked increase (P less than 0.001) in plasma protein concentration. Seven of the 10 horses developed signs of neuromuscular excitability, as evidenced by muscle fasciculations, and 5 of the horses developed diaphragmatic flutter. Hypernatremia was transiently induced, but it resolved as the horses were allowed access to water. The alkalosis induced by furosemide and NaHCO3 was profound and persisted for a 24-hour period and was associated with marked hypochloremia and hypokalemia. Partial replacement of the electrolyte deficits and correction of the metabolic alkalosis was attempted, using 1,000 mEq of NaCl or KCl given as an isotonic solution via nasogastric tube. In the KCl-treated group, there was a prompt and significant decline in venous blood pH and bicarbonate concentration (P less than 0.001) accompanied by a significant increase in plasma potassium concentration (P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551203 TI - In vitro testing of a potential retroviral vector for producing transgenic livestock. AB - The amphotropic murine leukemia virus (MLV) has been shown to infect mammalian species other than mice. If this virus infects and expresses genes in cells of livestock species (cattle, sheep, and pigs) it has potential for use as a vector to produce transgenic livestock. Because the gene-injection technique for producing transgenic animals is inherently inefficient, our laboratory was interested in identifying or constructing retroviral vectors capable of infecting livestock embryos. The infectivity of an amphotropic MLV-based vector for ovine, bovine, and porcine cells was tested. Experiments were also conducted to test the ability of the amphotropic MLV promoter, compared with known strong promoters, to express genes in cells from these species. Results indicated that amphotropic MLV infects and expresses genes efficiently in porcine cells and is, therefore, a potential vector for producing transgenic pigs. Infection was not detected in cells from adult bovine and ovine species; however, low levels of infection, with subsequent gene expression, were detected in cells derived from bovine embryos. PMID- 2551205 TI - Resolution of impaired pulmonary function and pulmonary hypertension after phorbol ester administration in rabbits. AB - To assess metabolic functions of the pulmonary circulation during lung injury and subsequent recovery from injury, we measured angiotensin-converting enzyme (ACE) activity by means of benzoyl-phenylalanyl-alanyl-proline (BPAP) hydrolysis and 5 hydroxytryptamine (5-HT) removal in vivo in three groups of anesthetized rabbits. One group was treated with 30 micrograms/kg/day phorbol myristate acetate (PMA) intravenously 10 times over 14 days (PMA group). A second group received the same PMA treatment but was not studied until 14 days after the last treatment (PMA/recovery group). A third group was treated with vehicle alone. At the end of PMA treatment, rabbits had an elevated pulmonary artery pressure and depressed ACE activity, expressed as the ratio Vmax/Km. Decreased Vmax/Km for ACE was due to a significant reduction in apparent Vmax for BPAP (control = 235 +/- 37, PMA = 139 +/- 12 nmol/s). Km was unchanged (control = 25 +/- 4, PMA = 31 +/- 7 microM). Uptake of 5-HT was unaffected by PMA treatment. After 2 wk of recovery (PMA/recovery group), pulmonary hypertension had resolved. In this group, Vmax for BPAP hydrolysis was not significantly different from control (280 +/- 18 nmol/s), but Km was significantly increased (48 +/- 5 microM). We conclude that repeated exposure of rabbits to PMA results in lung injury manifested as depressed pulmonary ACE activity and pulmonary artery hypertension. Although much of these alterations were reversible within 2 wk after discontinuing PMA, an increase in apparent Km of ACE may be a more persistent alteration in vascular endothelial cell dysfunction. PMID- 2551204 TI - Four-times-a-day dosing frequency is better than a twice-a-day regimen in subjects requiring a high-dose inhaled steroid, budesonide, to control moderate to severe asthma. AB - Fifty-three adult asthmatic subjects requiring 800 micrograms or more of inhaled beclomethasone dipropionate were enrolled in a double-blind parallel group study of 6-month duration to compare the efficacy and side effects of inhaled budesonide in doses of 800, 1,200, and 1,600 micrograms given two or four times a day (BID or QID). After a two-week observation period to establish baseline, subjects were given the same dose of budesonide as they would be for beclomethasone dipropionate; however, the frequency regimen (BID or QID) was randomly allocated. Subjects were asked to fill in diary cards describing their asthmatic symptoms, need for medication, and throat symptoms. They were assessed by a physician every 4 wk, at which time spirometry was performed. Throat swabs were done at every other monthly visit. Cortisol and response to cortisol after synthetic ACTH injection were assessed at the beginning and end of the trial. Thirty-six subjects, half in each group, completed the study. Those on the BID regimen had almost twice as many days with nocturnal asthma and cough and almost three times as many days with disability due to asthma. There were also twice as many relapses in the BID regimen as judged by the clinician. These relapses mainly occurred toward the end of the study. The maximal daily swings in peak expiratory flow rates were slightly greater in the BID group, although this difference was not physiologically significant. Spirometry, cortisol secretion, and the response after synthetic ACTH injection were not significantly different in either group from the beginning to the end of the study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551206 TI - [The versatile midface degloving approach. Re-updating the method, apropos of a case]. AB - A recurrent adenoid cystic carcinoma of the hard palate extending to the maxillary sinus was resected through the versatile midface degloving approach. The surgical technique is discussed, and advantages and limits of the procedure are analyzed. The authors recommend this approach for midface tumors which avoids a facial scar and which sounds oncological. PMID- 2551207 TI - Red blood cell scan in cavernous hemangioma of the larynx. AB - Cavernous hemangioma of the larynx is an uncommon, difficult-to-diagnose vascular tumor for which there is no significant imaging literature to date. The possibility of improved diagnosis through RBC scanning might obviate injudicious biopsy and potential hemorrhage within the airway. Utilizing the radionuclide RBC scan, which labels the patient's own RBCs initially with cold pyrophosphate, and subsequently with technetium 99m as pertechnetate, we have identified successfully four patients with cavernous hemangioma of the larynx. All presented with a supraglottic mass involving at least the aryepiglottic fold and arytenoid region unilaterally. This report describes our satisfactory diagnostic imaging experience with the radionuclide RBC scan and suggests both its imaging specificity and its role in the management of this lesion. PMID- 2551208 TI - Hydroxylapatite for laryngotracheal framework reconstruction. AB - Hydroxylapatite plates and rings were employed for reconstruction of the laryngotracheal framework in 12 patients. The cause of the framework defect was surgery for stenosis in nine cases and removal of malignancy in the other three. The trough method associated with a two-stage procedure was tolerated in all 12 cases. Hydroxylapatite plates or rings were used to reconstruct the framework during the second stage. The implant was well taken in all cases, without any infection or rejection. Nine of the 12 cases resulted in excellent airways. PMID- 2551209 TI - [Syringoma of the vulva underneath a seborrheic wart]. PMID- 2551210 TI - [Cutaneous tumors and adnexal cysts in children]. PMID- 2551211 TI - [Syringocystadenomatosis papilliferus: a case with a thoracobrachial site]. PMID- 2551212 TI - Histochemical study of the biphasic cellular pattern of tubular adenoid cystic carcinoma with azophloxine, amidoblack and acid blue. AB - A tubular adenoid cystic carcinoma of the soft palate was studied histochemically using tannic acid-phosphomolybdic acid-Azophloxine (TPAzf), tannic acid phosphomolybdic acid-Amidoblack (TPA) and tannic acid-phosphomolybdic acid-Acid Blue techniques. At the periphery of some tumor nests a few spindle cells were noted stained intensely red, blue-green and dark blue with TPAzf, TPA and TPAcB respectively. The apical portion or the whole cytoplasm of the cuboidal, eosinophilic cells that lined the central lumina of most neoplastic nests showed a similar tinctorial behavior. On the basis of their location and staining properties, the spindle cells were recognized as myoepithelial. On the other hand, the tinctorial behavior of eosinophilic cells was attributed to the dye binding by a conspicuous terminal bar-terminal web system or by tonofibrils accumulated in their cytoplasm as a result of squamous metaplasia. An inverse relation between the presence of myoepithelial cells at the periphery of tumor nests and the increasing thickness of terminal bar-terminal web system or the appearance of numerous tonofibrils in the eosinophilic cells was also noted. PMID- 2551213 TI - Host response to papilloma virus in focal epithelial hyperplasia. AB - The ultrastructural features of the lesions of four FEH patients disclosed changes in the nuclei interpreted as a premature disintegration of the nucleoli and the presence of papilloma-like viruses. Changes in ribosome number and distribution, glycogen content and formation of vesicles within the cytoplasm of FEH keratinocytes were observed and compared with normal epithelia. The observed cell changes were explained as specific host response due to HPV-13. PMID- 2551214 TI - Fluconazole (UK-49,858) treatment of candidiasis in normal and diabetic rats. AB - Fluconazole (UK-49,858), a new oral bistriazole antifungal agent, was compared with amphotericin B in the treatment of established systemic infection with Candida albicans in normal and diabetic rats. In normal rats, oral fluconazole at 10 mg/kg per day for 7 days reduced Candida colony counts in the kidneys and livers as well as amphotericin B did and was nearly as effective as amphotericin B in a 21-day treatment trial. There was no further reduction in Candida colony counts when normal rats were treated with fluconazole at 40 mg/kg twice a day for 7 days. In streptozotocin-induced diabetic rats, fluconazole at 20 mg/kg per day for either 7 or 21 days compared favorably with amphotericin B in efficacy. Results of our study suggest that oral fluconazole may be useful in the treatment of established disseminated candidiasis in normal as well as diabetic hosts. PMID- 2551215 TI - In vitro susceptibilities of yeasts to a new antifungal triazole, SCH 39304: effects of test conditions and relation to in vivo efficacy. AB - We used six candidal strains (two Candida albicans and one each of four other species) to study the effects of test conditions on the activity of SCH 39304 compared with that of fluconazole in broth macro- and microdilution assays. Increasing the inoculum from 10(2) to 10(5) yeasts per ml raised the MICs for all isolates up to greater than 512-fold. In contrast, results with a 50% turbidimetric endpoint (50% inhibitory concentration; IC1/2) varied no more than twofold. Similar effects were seen with fluconazole, and both drugs were found to have an associated delay in onset of action. Acidity was found to increase both MICs and IC1/2s. Other effects were observed among four synthetic media, but a consistent pattern was not identified. Incubation temperatures of 37, 35, and 30 degrees C yielded equivalent results. Broth microdilution IC1/2s against most of 40 isolates of C. albicans were 0.31 microgram/ml +/- fourfold for SCH 39304 and 0.16 microgram/ml +/- twofold for fluconazole. Treatment of experimental candidiasis in rats with SCH 39304 and fluconazole resulted in 50% effective doses of 0.33 and 0.49 mg/kg per day, respectively. In contrast, another C. albicans isolate, previously identified as resistant to other azoles, had IC1/2s of 20 micrograms of SCH 39304 per ml and, in vivo, a 50% effective dose of 2.25 mg/kg per day. We conclude that the in vivo efficacy of SCH 39304 correlates with MIC results when broth macrodilution testing is performed with a small inoculum and with IC1/2 results which are independent of inoculum size. PMID- 2551216 TI - [Combination chemotherapy with bronchial arterial infusion (BAI) and systemic chemotherapy in advanced non-small cell lung cancer]. AB - Five patients of advanced non-small cell lung cancer with atelectasis and/or poor performance status were treated with combination MVP therapy consisting of bronchial arterial infusion (BAI) using cisplatin (50 mg) and simultaneous systemic MVP therapy. The tumor size reduced more than 50% in three patients, and less than 25% in one patient. In another patient the tumor size was not measurable. Four of five patients showed disappearance of atelectasis, and three of four patients showed improvement of performance status. Average time taken was 13 and 17 days, respectively. We expect that combination MVP therapy has beneficial effects not only for relieving local symptoms and improving performance status within shorter period, but also achieving a better prognosis. PMID- 2551217 TI - [Intra-arterial infusion chemotherapy of locally advanced and recurrent breast cancer]. AB - A total of 15 cases of advanced and recurrent breast cancer of more than 3-year duration were subjected to continuous intra-arterial infusion in our department, and its therapeutic effect and the judgement were examined. Using subclavian artery and internal thoracic artery for the primary cases and the latter artery for the recurrent cases for infusion, continuous administration of 5-FU and one shot injection of ADM and MMC were performed twice a week. 99mTc-angiography was effective for the identification of arterio-controlled areas. Under the judgement criteria for therapeutic effects according to the breast cancer handling agreement, CR, PR, NC and PD were noted in 1, 8, 5 and 1 cases, respectively, with a 60% rate of effectiveness. Judgement of effects by 201Tl-chloride scintigraphy showed some correlation with the histological judgement, with its apparent effectiveness. Examination of the remote results revealed 3-year survival rate of 83.3% and 3-year healthy survival rate of 66.7% for the primary cases. Two of the 3 recurrent cases died at 2 and 11 post-infusional months, respectively, but 1 showing CR is still alive 4 years and 10 months, post infusionally. PMID- 2551218 TI - [Intra-arterial infusion chemotherapy in non-resectable pancreatic cancer using angiotensin-II and implantable drug delivery system]. AB - Over the past 6 years, we have treated 25 cases of pancreatic cancer, 6 cases of cholangioma in pancreas-head and 3 cases of cancer in duodenal papilla (2 cases Stage I, 5 cases stage II, 2 cases stage III, 25 cases stage IV). Twelve cases (10 unresectable cases, 1 hepatic metastasis case, 1 recurrent case) were treated with intra-arterial infusion chemotherapy using implantable Drug Delivery System, combined with angiotensin-II to increase the concentration of anti-cancer agents in cancer tissue. Twenty-four cases (70%) died in less than one year, so operation is not effective except for curative resection of cholangioma and duodenal papilla cancer. But exploratory laparotomy or inoperable cases given intermittent transcatheter arterial infusion chemotherapy (5-FU + ADM + MMC + angiotensin-II), showed favorable results (decrease of tumor size and pain in 2 cases; recanalization of obstruction in choledochus of 1 case). Especially trans femoral or left subclavian arterial catheterization proved to be effective therapy for possibly giant or recurrent inoperable pancreatic cancer and hepatic metastasis. Using the drug delivery system, the technical approach to arterial infusion therapy and angiography has been readily undertaken. Quality of life has been improved, and course observation of the patient has been possible by imaging diagnosis and multidisciplinary treatment for advanced pancreatic cancer. PMID- 2551219 TI - [Local immunotherapy of breast cancer: a case of advanced breast cancer improved by combined local injection therapy with OK-432 and rIL-2]. AB - An effective immunotherapy for breast cancer has yet to be established. We have recently experienced an inoperable case of advanced breast cancer due to mental disorder. Combined local injection therapy with OK-432 and rIL-2 was performed in addition to non-surgical multi-disciplinary treatment, with obvious therapeutic effects. The case is a 67-year-old female, who had a 5.0 x 4.7 cm tumor in the area A of the left mammary gland. Aspiration cytologic examination revealed class V breast cancer. Besides systemic chemotherapy, local injection therapy with OK 432 2.0 KE on day 1 and rIL-2 500 U on day 3 was performed. Upon starting local injection therapy, a rapid reduction was noted in the tumor diameter (reduction rate, 75.5%). Tumor makers of CEA, CA 15-3 also showed obvious diminution. Although the dose and interval of administration remain to be further examined, the combined local injection therapy with OK-432 and rIL-2 was suggested to be an effective local immunotherapy for breast cancer. PMID- 2551220 TI - [New approach for unresectable primary liver cancer--intra-arterial combination chemotherapy during and after operation with local injection of ethanol]. AB - From 1976 to 1988, hepatic arterial infusion chemotherapy was performed in 52 patients with unresectable hepato-cellular carcinomas. Forty-nine (94%) out of 52 patients were stage III or stage IV. Thirty-five patients underwent continuous infusion of 5-FU and MMC employing chronometric infusion pumps. Mean survival of this group was 9 months. Six patients survived more than 12 months; the longest survivor died of different causes 40 months after surgery. Another patient lived for 30 months. In 10 patients, hepatic arterial ligation was performed in combination with infusion chemotherapy in the hepatic artery. Mean survival of these patients was 14 months. Four out of the 10 patients survived over 12 months. Recently, in 7 patients whose liver tumors were less than 4 cm in diameter and/or whose tumors were multiple and had liver disfunction, ethanol injection into the tumors during laparotomy was performed in combination with hepatic arterial infusion chemotherapy. In these 7 patients, an injection port system was implanted subcutaneously for intermittent infusion chemotherapy of MMC + 5-FU and ADM + lipiodol. Mean survival in this group was 14 months. One of 7 patients died 21 months after operation, but the others are alive at this writing. Five of these 6 patients have survived more than 12 months. AFP levels decreased in 3 patients after these treatments. In one case, AFP decreased to normal range for 18 months. These results suggested that intraoperative ethanol injection therapy into the tumors combining with hepatic arterial infusion chemotherapy may be of value for unresectable smaller liver carcinomas, since this method is technically easy and can assure injection of ethanol into the tumors during laparotomy. PMID- 2551221 TI - [Usefulness of subcutaneously implanted reservoir for postoperative therapy in hepatocellular carcinoma and liver metastases of colorectal carcinoma]. AB - Hepatectomy has been a treatment of choice for hepatocellular carcinoma and metastatic liver carcinoma. Recurrence in residual liver after hepatectomy is clinically a serious problem. Since 1987, postoperative hepatic arterial infusion chemotherapy using subcutaneously implanted reservoir has been undertaken to improve the prognosis after hepatectomy in hepatocellular carcinoma and liver metastasis of colorectal carcinoma. The indications for reservoir implantation were determined for high-risk cases in hepatocellular carcinoma and all cases in liver metastasis. The tip of a catheter was placed at the root of the common hepatic artery via gastroduodenal artery. Lipiodol-ADM was injected for hepatocellular carcinoma every 2 months and MMC-5-FU was injected for liver metastasis of colorectal carcinoma every one or two weeks. Complications of this procedure in every 2 cases of reservoir infection proved to be catheter obstruction and hepatic artery obstruction. In the process of this treatment, we observed 3 recurrences in residual liver of hepatocellular carcinoma and one case of peritoneal dissemination and 3 recurrences in residual liver of liver metastasis of colorectal carcinoma. All are still alive. PMID- 2551222 TI - [Hepatic arterial infusion chemotherapy of hepatocellular carcinoma]. AB - In 24 cases of unresectable hepatocellular carcinoma, we performed hepatic arterial catheterization and intra-arterial infusion chemotherapy. Adriamycin (ADM), Mitomycin C (MMC), 5-FU and Lipiodol (LPD) were administered an average of 13.5 times over a mean period of 106 days. Except for 5 unevaluable cases, there were 0 CR, 5 PR, 4 MR, 7 NC and 2 PD cases, for an efficiency rate of 27.8%. Complications thought to be due to the catheter included catheter blockade in 1 case (4.3%) and dermal infection of insertion site in 3 cases (13.0%). As for the results of follow-up study, one-year survival rate with this therapy was 47.8%, which compares favorably with a one-year survival rate of 30.0% in 30 cases treated only with TAE. From the above results, hepatic arterial infusion chemotherapy can be repeatedly performed on an outpatient basis, and it is considered to be a useful therapeutic method for treating unresectable hepatocellular carcinoma. PMID- 2551223 TI - [Superselective transcatheter-arterial-chemo-embolization in uterine cervical cancer]. AB - Transcatheter-arterial-chemo-embolization (TACE) was performed on 7 cases of uterine cervical cancer from T1b to T3b. The purpose of this approach is to stop the uterine bleeding caused by malignancy and disrupt the cancer cells under the feeding arteries. TACE by Seldinger's method was undertaken selectively on both uterine arteries with suspension of Lipiodol, Adriamycin and Gelfoam. In each case, the bleeding stopped soon after the operation, and the lesion began to necrose within a few days. Though the primary effects were confirmed, complete remission could not be expected, so radiation therapy was performed. The longest survival among these patients was over 6 years. It should be noted that the other major advantage claimed for TACE is to decrease the dose of radiation. PMID- 2551224 TI - [Enhanced effect of hepatic arterial infused anticancer drugs on rabbit VX-2 carcinoma by means of combination with Lipiodol. Adriamycin versus Lipiodol adriamycin sandwich method and adriamycin suspended in a Lipiodol]. AB - To increase drug delivery to a hepatic malignant tumor, various tissue contents and serum levels of adriamycin (ADR) were evaluated in comparison with different intraarterial injection methods of oily or aqueous solution. VX-2 tumor was implanted in the rabbit liver. ADR was infused through the hepatic artery by the following three methods: Group 1: ADR with physiological saline; Group 2: ADR administered between lipiodol injection; and Group 3: urographin-dissolved ADR suspended in lipiodol. ADR concentrations of the tumor, liver, heart, kidney or bone marrow were compared between these three groups one hour after ADR administration. Tumor uptake and ADR concentration ratio of tumor to liver were higher in Group 3 than in the other two groups. ADR concentration ratio of tumor to liver was higher in group 2 than in group 1. Cardiac and renal uptake in group 3 was less than in group 1 and 2, and so infusion of ADR suspended in a lipiodol may serve to reduce side effects of ADR, especially cardiotoxicity. PMID- 2551225 TI - [Hepatic arterial infusion chemotherapy and hyperthermia with degradable starch microspheres in primary and metastatic liver malignancies]. AB - In non-resectable liver malignancies, concurrent administration of degradable starch microspheres (DSM) and anticancer drugs via hepatic artery has been suggested as a method to increase the concentration of drugs in tumor tissue. DSM also has been known to increase the temperature of tissue when administered at the time of hyperthermia. In the light of these findings we have studied the effect of hepatic arterial infusion of 5-FU and mitomycin C and local hyperthermia in combination with hepatic arterial flow arrest with DSM for the treatment of hepatoma in 11 patients and metastatic liver cancer in 38 patients. Of the 8 patients having hepatoma with increased AFP, all the patients showed a decrease of AFP following the therapy with an average ratio of 65% decrease. Of the 33 patients with hepatic metastasis with increased CEA, 32 patients (96%) showed a decrease of CEA following the therapy (control group with infusion chemotherapy and hyperthermia without DSM: 58%) with an average decrease ratio of 59% (control group: 43%). Of the 26 patients with increased CA 19-9, 22 patients (84%) showed a decrease of CA 19-9 (control group: 75%) with an average decrease ratio of 52% (control group: 29%). This pilot study suggests that the concurrent hepatic arterial infusion of 5-FU, mitomycin C and DSM with local hyperthermia may have the potential to improve selective regional drug effect. PMID- 2551226 TI - [Continuous arterial infusion therapy of recombinant interleukin-2 and cyclophosphamide in hepatoma and liver metastasis]. AB - The continuous arterial infusion of only recombinant interleukin-2 (rIL-2) was used in 9 patients with unresectable liver metastasis of colorectal or biliary cancer. This therapy was undertaken for the activation of tumor infiltrating lymphocytes (TIL), but was not effective. Then we tried continuous arterial infusion of rIL-2 and low-dose cyclophosphamide (CY) for liver metastasis. 3 of the patients who received rIL-2 and CY showed partial remission of disease. This therapy was possibly effective because of the activation of TIL and the suppression of tumor infiltrating suppressor lymphocytes. PMID- 2551227 TI - [Limitation of oily chemoembolization against hepatocellular carcinoma and arterioportal segmental chemoembolization]. AB - Emulsion of adriamycin and lipiodol added with Gelfoam has been used for the transcatheter oily chemoembolization against hepatocellular carcinomas. This therapy resulted in significantly increased cumulative survival as compared with chemoembolization therapy using anti-cancer drugs and Gelfoam. However, it was revealed to have the following limitations: Insufficient efficacy against micrometastases and extracapsular invasion; impossible to perform sufficient treatment for patients with severe hepatic disorders; and there were cases in which lipiodol was hardly retained. In order to overcome the limitations described above, we have performed segmental hepatic chemoembolization simultaneously from the hepatic artery embolization simultaneously from the hepatic artery and portal vein. This method works on the basis of the fact that excess amount of lipiodol emulsion infused from the hepatic artery flows into the portal vein. Gelfoam embolization is also added in this method. Though subjects appropriate for this newly developed method are restricted, all the tumors in the objective segment of the liver can undergo necrosis. The efficacy is comparable to the effects of hepatectomy. In addition, since the other segments of the liver are able to be kept in reserve, this method is able to be applied to patients with severe hepatic disorders. PMID- 2551228 TI - [Segmental arterio-portal chemo-embolization in hepatocellular carcinoma (cement therapy)]. AB - When a large amount of iodized oil (LPD) is selectively administered to targeted segments of the liver, the tumor vessels and sinusoids around the tumor are filled with LPD, subsequently regurgitating excessive LPD via the hepatic arterioportal communication into the portal branches surrounding the tumor. Segmental arterioportal chemoembolization in HCC with Doxorubicin-in-oil emulsion (DOE), which we named cement therapy, was performed in 18 patients with HCC. DOE was administered selectively into one segment division of Couinaud in ten patients, two segments in seven and three segments in one. Computed tomography and ultrasonography following the cement therapy revealed an overall response rate of 39%. The one-year cumulative survival rate was 80%, and that for three years was 33%. Three cases underwent segmental hepatectomy and these specimens revealed total necrosis of the main tumor and whole daughter nodules, followed by various degrees of hepatic parenchymal infarction. This cement therapy realizes the simultaneous chemoembolization via the artery and portal vein, and was considered an effective treatment for extra-capsular infiltration and daughter nodules of HCC nourished by the portal vein blood flow. Limited administration of DOE to the targeted segments enables transarterial oily chemoembolization (TOCE) without any serious side effects. PMID- 2551229 TI - [Intra-arterial and intraportal therapy combined with decollateralization in unresectable cholangiocellular carcinoma--a case report]. AB - A 45-year-old man was referred to our hospital due to obstructive jaundice by intrahepatic cholangiocellular carcinoma. At laparotomy, he was assessed as unresectable because of multiple foci in both lobes. Therefore, two catheters connected to implantable access devices were placed in both the hepatic artery and the portal vein following decollateralization using silicone rubber sheeting, in addition to choledochectomy and choledocho-jejunostomy. After the surgical procedure, he underwent chemoembolization twice using Lipiodol, cisplatin, and Gelfoam, and two intraportal infusions of cisplatin at a dose of 50 mg. Moreover, a total dose of 4 g of 5-FU and 20 mg of MMC was administered through the arterial and portal catheters, respectively. Administration of G-CSF was remarkably effective for severe thrombocytopenia, and leukopenia resulted from this active chemotherapy. He is still alive 7 months after the surgical procedure with the regression of the lesions. PMID- 2551230 TI - [Chemo-embolization therapy of unresectable liver metastases using implantable infusion port]. AB - Since 1987, 14 patients (10 colorectal, 3 gastric and 1 lung cancer) with unresectable liver metastases received intra-arterial infusion chemo-embolization therapy using implantable infusion port. All patients had more than one lesion in bilateral lobe (H2 and H3). Infusion catheters were placed in the proper hepatic artery through the gastroduodenal artery on laparotomy. Infusion ports were implanted in the subcutaneous tissue of the abdominal wall. Various kinds of chemotherapeutic agents such as MMC, ADR, THP-ADR, CDDP and 5-FU were injected with embolization material (DSM or Lipiodol), every 1 to 4 weeks at the outpatient clinic. Among 10 cases of H2 grade metastases, 1 CR and 3 PR (40% clinical response) were obtained. However, all 4 cases of H3 grade were judged PD. All patients except one with H2 grade metastases are still alive, but 3 out of 4 with H3 grade died within 7 to 11 months. Catheter occlusion was observed in 4 cases for 3 to 7 months. Infection around the port occurred in 1 patient. A patient with metastatic liver cancer was treated by intermittent bolus injection with MMC and DSM. Partial response was confirmed by CT and tumor markers. Histological response was demonstrated in the specimen obtained at partial hepatectomy. It is concluded that this treatment is variable to prolong the survival of patients with H2 grade metastatic liver cancer, together with maintenance of the quality of life. PMID- 2551231 TI - [Stagnation of liver blood and effectiveness of intra-arterial chemotherapy in metastatic colorectal cancer]. AB - From the intraarterial infusion tube in the proper hepatic artery, 5-10 mCi of 133Xe (T1/2 = 5.3 days) was injected. The duration of blood stagnation (T1/2) in the tumor site was obtained from the washout curves in the tumor site and the nontumor site as the duration from the peak to the decrease to half value. The results indicated that the stagnation lasted 110.3 +/- 70.0 seconds in the tumor site, and 80.1 +/- 41.0 seconds in the non-tumor site, showing that the blood was more apt to be stagnant in the tumor site than in the non-tumor site. In the foci in which the stagnation in the tumor site (T1/2) was more than 100 seconds, the response was obtained for 6 (75%) out of 8 foci, which was higher than 3 (60%) out of 5 foci having abundant vascularity. In the foci in which the stagnation in the tumor site did not exceed 80 seconds, no response was obtained irrespective of the vascularity. From the above review, the intraarterial infusion method for the metastases of colorectal cancer was effective for the case which required a long time for washout of the anticancer agent at the tumor site. PMID- 2551232 TI - [Two cases of advanced breast cancer with distant metastasis showing long-term survival with multidisciplinary treatment including intra-arterial infusion chemotherapy]. AB - Two cases of advanced breast cancer are reported that have had no recurrence for more than five years after multidisciplinary treatment including intra-arterial infusion chemotherapy. The first case is a 62-year-old housewife who had brain metastases. The primary lesion and distant metastases were completely responsive to irradiation combined with chemoendocrine therapy. The other one is a 38-year old housewife with ovarian metastasis that was revealed after oophorectomy. In this case chemoendocrine therapy was added without irradiation after intra arterial infusion chemotherapy. These cases indicate that intra-arterial infusion chemotherapy can be recommended as a component of multidisciplinary treatment even in advanced breast cancer with distant metastasis. PMID- 2551233 TI - [A case of massive hepatoma which responded to SMANCS/Lipiodol regimen with intra arterial infusion]. AB - Transcatheter arterial chemotherapy (SMANCS/lipiodol) was applied to massive hepatoma, which had a high AFP 213,000 ng/ml, A-P shunt, tumor thrombosis and metastatic lung cancer. After 3 months, the AFP value reduced to 18 ng/ml, massive hepatoma and the A-P shunt disappeared, but AFP-negative nodular hepatoma recurred around initial hepatoma. Each time, we injected SMANCS/lipiodol to the recurring hepatoma. The therapy in the initial stage was not so effective. The portal vein was not observed in the initial stage, but appeared after the second dosage. Metastatic lung cancer was declining in the initial dosage and 23 months later disappeared after the third dosage. The massive hepatoma occupied entirely the rt. lobe of the liver. The patient lived for 4 years, had total admission periods of 190 days and could return to life in society. In this case, we considered that transcatheter arterial chemotherapy (SMANCS/lipiodol) had remarkable effects. PMID- 2551234 TI - [A case of recurrent hepatocellular carcinoma successfully treated by arterial chemoembolization in combination with local hyperthermia]. AB - We report a long-term survival case of recurrent hepatocellular carcinoma (HCC). A 40-year-old woman suffering advanced HCC underwent right hepatic tri segmentectomy. After the surgery, HCC recurred in the remnant liver and lung. An arterial infusion of Lipiodol emulsion of anti-cancer agents and regional hyperthermia therapy were repeatedly performed. Administration of FT and PSK were also started. After this combined therapy, AFP levels decreased and recurrent lesions in the liver as well as pulmonary metastases disappeared on image studies. The patient is doing well 4 and half years later with no evidence of recurrence. PMID- 2551235 TI - [A long-survival case of hepatocellular carcinoma treated by intra-arterial chemotherapy and immunotherapy]. AB - Hepatic resection is generally considered to be superior to any other therapeutic procedures for hepatocellular carcinoma (H.C.C.). However, the resectability of the patients who have HCC. with liver cirrhosis is still low, and surgery is appropriate in only a minority of patients. Although some successful reports of intra-arterial chemotherapy for HCC. have been documented, most of the therapeutic effects are transient and the survival rate is not satisfactory. This report is of a rare case, that of a long-term survivor with HCC treated by intra arterial chemotherapy and immunotherapy. A 66-year-old man, with a 10-year history of liver cirrhosis was admitted to The Center for Adult Diseases, Osaka, after detection of a tumor in the right lobe on US. On admission, serum AFP was within normal range, HBs-Ag was negative, and ICG-R 15 was 20.8%. On hepatic angiogram, a hypervascular tumor (6 cm in size) was recognized in the middle of the right lobe. He was assessed as unresectable because of insufficient reserve capacity, and the catheterization of the hepatic artery for intra-arterial chemotherapy and the injection 35 KE of OK-432 into the tumor were carried out under laparotomy. After the procedure, the patient was treated by intra-arterial infusion of doxorubicin (ADR) at a total dose of 150 mg and 5-FU in total dose of 25 g, with a hypodermic injection of OK-432 at a total dose of 161 KE. Hepatic angiography, carried out one year after the procedure, disclosed no foci in the liver. The duration of complete remission continued more than 5 years. The patient eventually died of intrahepatic recurrence, but he lived for 7 years and 3 months after the catheterization. PMID- 2551237 TI - [Assessment of three-year survival cases with unresectable hepatocellular carcinoma treated by arterial chemotherapy]. AB - The survival rate was studied in 218 patients with unresectable hepatocellular carcinoma (HCC) in whom arterial chemotherapy was performed by one-shot injection of anticancer agents (one-shot therapy) was studied. Tumor factors and clinical features of 6 out of 218 cases with more than 3-year survival, were also studied. The following results were obtained; (1) One-year survival rate of the cases which were compatible with criteria for one-shot therapy in our department was 35.6%, and the three-year rate was 7.0%. On the other hand, one-year survival rate of the cases not compatible with same was 1.5%. The latter survival rate was worse than that for conservative care. (2) The longest survival time was 6 years and 5 months after one-shot therapy. (3) The tumor progression rate was above 40% in 4 out of 6 cases which survived more than 3 years, and tumor thrombus in the portal vein was noted in 3 cases. Clinical stage was I-II. (4) One-shot therapy was performed repeatedly after initial treatment in these 6 cases. In 3 of the 6 cases, TAE or percutaneous ethanol injection therapy was performed for recurrence or progression of HCC during observation. The results suggest that criteria for one-shot therapy and multidisciplinary treatment during observation are necessary. PMID- 2551236 TI - [Analysis of prognostic factors in patients with hepatocellular cancer treated by hepatic arterial infusion chemotherapy]. AB - Analyses of the prognoses of thirty-four patients with hepatocellular cancer, who were treated by hepatic arterial infusion chemotherapy and/or transcatheter arterial embolization (TAE) and/or hyperthermotherapy, were performed by multivariate analysis using Cox's proportional hazard model and generalized Wilcoxon test. In the multivariate analysis on the conditions of patients, nine of fifteen variables were associated with the prognosis of patients who received regional cancer chemotherapy. The variables are: sex, liver cirrhosis, esophageal varices, GOT, GPT, albumin and gamma-globulin. One of three variables was associated with the prognosis in the therapy analysis, and the variable is TAE. Significant differences in survival curves which were estimated by generalized Wilcoxon test were noted in age, portal vein invasion, ascites, GOT and TAE. From these results it is suggested that the conditions of patients with unresectable hepatocellular cancer must be carefully investigated before regional cancer chemotherapy and the good therapy effects is obtained by hepatic arterial infusion chemotherapy combined with transcatheter arterial embolization therapy. PMID- 2551238 TI - [Eight cases of non-resectable hepatocellular carcinoma surviving for more than 3 years after Lipiodol administration]. AB - From December 1982 to February 1986, selective regional cancer chemotherapy using Lipiodol plus anticancer drug (Lipiodolization) was prescribed for 87 patients with non-resectable hepatocellular carcinoma (HCC). Eight out of 87 cases survived for more than 3 years. The mean age of the 8 long survivors was 52.6. In the liver function, 2 cases were in Child A, 5 cases in Child B, and 1 case in Child C. The largest diameter of HCCs was 5.3 cm, and the number of the tumors was one in 6 cases, two in one case, and three in one case. No vascular invasion was detected on hepatic angiography. These findings suggested that long survivors in lipiodolization are not in far advanced HCCs. For these 8 cases, lipiodolization was repeatedly prescribed from two to 6 times. The largest amount of adriamycin was 192 mg/case. The longest survivor has lived for 5 years and 6 months after first lipiodolization. After lipiodolization, one tumor vanished, and three tumors decreased in size. Although seven tumors increased in size after lipiodolization, the tumor doubling time of seven tumors were 64, 265, 313, 317, 350, and 1943 days (average 539 +/- 639 days). It is possible that lipiodolization remarkably inhibited tumor growth. Nevertheless, in 5 out of 7 cases, the daughter lesions increased in size and number, although the main tumors grew slowly. Lipiodolization was less effective for newly arising daughter lesions than main lesions. PMID- 2551240 TI - [Distribution of pulsed intra arterial infusion chemotherapy in hepatic carcinomas]. AB - Evaluation of "Gianturco-Wallace chemotherapy pulser," which was developed to produce a more homogeneous drug distribution of the tumors in intra-arterial infusion chemotherapy, was assessed by comparative study of pulsed and nonpulsed arterial radionuclide infusion using Tc-99m pertechnetate for 18 cases of hepatic carcinomas (11 cases of hepatocellular carcinomas and 7 cases of metastatic hepatic carcinomas). Tc-99m pertechnetate, 740 MBq (20 mCi) diluted with saline (30 mL) was infused with or without pulse through the catheter into the hepatic artery at a rate of 1mL per minute. The intrahepatic dynamic radionuclide distribution was analyzed by the time activity curves of ROIs in the tumor and nontumor areas. Pulsed infusion interrupted laminar flow and produced more homogeneous radionuclide distribution in the liver, and combination of pulsed and nonpulsed infusion also produced better radionuclide distribution in the areas of the tumors. This method using Tc-99m pertechnetate was very useful as a simulation to determine the dynamic drug distribution of the tumor and non-tumor region in intraarterial infusion methods. PMID- 2551239 TI - [Usefulness of measurement of hepatic arterial blood flow during surgery to insert an arterial reservoir in unresectable hepatocellular carcinoma]. AB - In a case of hepatocellular carcinoma, complicated with celiac arterial constriction, for which resection was impossible, we measured the blood flow in the hepatic artery using an ultrasonic, transit-time rheometer during surgery, and consequently inserted a reservoir safely. The measurement of the blood flow in this case demonstrated that the proper hepatic artery was provided with blood from superior mesenteric artery by the gastroduodenal artery and that the blood flow of the common hepatic artery was directed to the root part of the celiac artery. Moreover, because blood flow through the proper hepatic artery was greater while clamping the common hepatic artery than the gastroduodenal artery, cannulation was carried out from the common hepatic artery. Postoperatively, when recombinant interleukin-2(rIL-2) was infused from the reservoir, the tumor disappeared on imaging pictures, and the level of AFP (which was high preoperatively) was reduced, which was thus judged as a complete response. These results indicated the usefulness of measurement of blood flow in surgery of inserting a reservoir in cases with abnormal blood circulatory dynamics. PMID- 2551241 TI - [Clinical study of the drug delivery in metastatic liver cancer using bromodeoxyuridine]. AB - As the model of an anti-cancer agent acting at DNA synthesizing phase, BrdU was infused for metastatic liver cancers into the portal vein and/or the hepatic artery. After administration, the liver tumors were resected, and immunohistochemical staining was performed using anti-BrdU monoclonal antibody. Two of 6 portal group tumors were stained. All 4 arterial and 3 arterio-portal group tumors were stained. Therefore, it was proved that metastatic liver tumors had both arterial and portal blood supply. But areas to which BrdU was delivered were only peripheral, and the inmost area was 2 mm from the tumor surface. After the intra-arterial infusion of BrdU suspended in lipiodol, the delivery of BrdU was highly enhanced and 4 of 6 tumors were stained even to the deepest areas. However, one tumor of which the major part was necrotic and one tumor which produced mutin were not stained at all. It was interesting that after intra arterial administration of BrdU suspended in lipiodol, BrdU was also found in the cytoplasm of normal liver cells. PMID- 2551242 TI - [Studies on drug release from anti-cancer drug suspended Lipiodol]. AB - In case of an arterial infusion chemoembolization therapy for primary or metastatic liver cancer, gradual release of the anti-cancer drug from lipiodol is a very important factor for a higher drug concentration in the tumor and for longer contact. We studied basic points about what kind of drug form has a gradual drug release. We prepared 3 forms of drugs. (1) Powder form: Powder of ADM, MMC and CDDP was suspended in lipiodol with ultrasonic suspender. (2) URO form: ADM and MMC were dissolved with Urografin and mixed with lipiodol. (3) Surfactant form: ADM and MMC were dissolved with water and then mixed with lipiodol using surfactant. We put lipiodol suspension into physiological saline and then stirred water at 100 rpm with the paddle method, measuring drug release from the suspension or emulsion. Powder form had a lowest drug release. In clinical trials, we administered intra-arterially (1) ADM, MMC dissolved with physiological saline water as usually used (physiological saline water form) (2) Powder form; (3) URO from; (1) CDDP solution as usually used was administered; (2) Powder form. Then we studied the changes of serum concentration of ADM, MMC and CDDP. The results indicated that powder form had the lowest drug release. Thus, the water-soluble anti-cancer drugs ADM, MMC and CDDP should be used in powder form. PMID- 2551243 TI - [The study of transcatheter arterial chemo-lipiodol administration in liver metastasis of colorectal cancer]. AB - Transcatheter hepatic arterial chemo-lipiodolization (TAC), using totally implantable reservoir, was performed for the treatment of liver metastasis from colorectal cancers in three cases of H1 (metastasis in one lobe only, n = 10), in 3 cases of H2 (a few scattered metastases in both lobes, n = 7), and in 2 cases of H3 (numerous metastases in both lobes, n = 9). We performed TAC in H1 cases after resection of liver tumor. We found a recurrence on lipiodol CT 2 months later in 1 case of H1. The metastatic tumor responded to TAC in two patients of H2 (1 case completely), but no response was observed in H3 cases. The present study suggested the effectiveness of TAC in cases H1 and H2, but further study is needed for cases of H3. PMID- 2551244 TI - [Prognosis of intra-arterial chemo-embolization in metastatic liver cancer]. AB - We treated 63 patients (pts) suffering from metastatic liver cancer with intra arterial infusion chemotherapy, and analysed 44 of their for survival since the first treatment with regard to the primary foci of cancer and the method of intra arterial therapy. Via the superficial femoral artery, we performed superselective hepatic catheterization by Seldinger's method. Three types of intraarterial therapy were used: Gelfoam embolization with mitomycin-C (MMC) in 12 pts (GS TAE), capillary chemo-embolization with MMC-Lipiodol emulsion in 28 pts (LP-TAI) and "one-shot" slow infusion of MMC or cisplatinum in 4 pts. Fifty-percent survival was 189 days in pts with metastases from colo-rectal cancer (n = 20), 109 days from gastric cancer (n = 9), 100 days from pancreatobiliary cancer (n = 5) and 240 days from breast cancer (n = 7). More than one-year survival was obtained in 13 out of the 40 pts (32.5%). Survival of 12 pts, treated with GS-TAE regimen, was not significantly superior to that of 28 pts with LP-TAI regimen. Hence, we conclude that LP-TAI is the treatment of choice in chemo-embolization for unresectable liver metastases, because it causes less damage to the hepatic arterial beds, and facilitates repeat intraarterial therapy in these pts. PMID- 2551245 TI - [Adjuvant surgery to enhance therapeutic effects of chemo-embolization in advanced hepatocellular carcinoma]. AB - Different kinds of adjuvant surgery were carried out in three patients with advanced hepatocellular carcinoma (HCC) to enhance the therapeutic effects of chemo-embolization (TAE). The first patient, who had multiple foci localized in the right lobe, underwent ligation of the right portal vein in order to eliminate the small intrahepatic spreads and to protect against transportal metastasis to the left lobe. Afterward, TAEs were repeated a total of 5 times. The patient is still alive 24 months after the surgery with partial remission of the lesions. The second patient with severe cirrhosis had received TAE twice. Since one of the tumors, which existed in the hepatic hilus, did not respond to TAE, only the non respondent tumor was removed surgically. This patient remains alive 28 months after the surgery with complete remission of the lesions. In the third patient, repeated TAEs resulted in collateral feeding arteries. Permanent decollateralization using silicone rubber sheeting was carried out following ligation of collaterals. Thus adjuvant surgery to enhance the therapeutic effects of TAE may be a significant palliative treatment for HCC. PMID- 2551246 TI - [Selective anticancer effects of intra-arterial infusion of 3',5'-dioctanoyl-5 fluoro-2'-deoxyuridine (TT82) in hepatocellular carcinoma]. AB - 3'-5'-Dioctanoyl-5-fluoro-2'-deoxyuridine (TT 82), one of the lipophilic prodrugs of 5-fluoro-2'-deoxyuridine mixed with Lipiodol (LPD), was injected into the hepatic artery before hepatic resection in 4 patients with hepatocellular carcinoma. Later, the anti-cancer effects of the drug were evaluated mainly by the examination of resected specimens. The serum alpha-fetoprotein levels of three patients in whom the values were over normal range fell to less than 50% of pretreatment values from 7 to 14 days after the injections. In pathological observation of the specimens, the necrosis rates of main tumors were 100%, 70%, 30% and 0%, respectively. In three patients whose main tumors showed complete or partial necrosis, selective depositions of LPD in the tumors were observed on soft X-ray photographs. The concentrations of TT 82 and its metabolites in the tissues of LPD-depositing areas were markedly higher than those in tissue of the regions without LPD deposits. These findings suggested that TT82-Lipiodol mixture remained selectively in the cancerous tissues and exhibited anticancer effects following injection into the hepatic artery. PMID- 2551247 TI - [Intra-arterial injection of cisplatin suspension in Lipiodol (LPS) in the treatment of hepatocellular carcinoma]. AB - Cisplatin suspension in Lipiodol (LPS) was prepared for the treatment of hepatocellular carcinoma by intra-hepatic arterial injection. In a rabbit liver cancer model, concentrations of cisplatin in tumor were more than 20 times higher than those in a nontumorous part of the liver at 5 min after LPS injection into the hepatic artery. Cisplatin at high concentrations was detected at 7 days after injection. The concentrations in other organs were lower except in the gall bladder. In clinical trials for 71 patients with hepatocellular carcinoma, partial response was observed in 33 cases (46.5%) and minor response in 20 cases (28.2%). The survival rate was 77% at 6 month and 55% at one year. Although fever, nausea, vomiting and epigastralgia were observed as side effects, these were temporary. Acute gastroduodenal mucosal lesions, cholecystitis, pancreatitis, delayed jaundice and hepatic encephalopathy were observed as complications and super selective cannulation was necessary for their prevention. PMID- 2551248 TI - [Chemo-embolization using degradable starch microspheres and regional hyperthermia in unresectable hepatocellular carcinoma]. AB - Antitumor effects and prognosis of the patients with unresectable hepatocellular carcinoma were investigated in the treatment of chemo-embolization using degradable starch microspheres (DSM) and regional hyperthermia. Twenty-six cases were treated with chemo-embolization using DSM alone (Group A) and eighteen cases were treated with combination of chemo-embolization and hyperthermia (Group B). Tumor regression rates over 50% were 42% (11/26) and 56% (10/18) in Group A and Group B, respectively. In the patients with tumors over 7 cm in diameter, no favorable response was obtained in Group A (0/11), but the response rate was increased to 56% (5/9) by the combination of hyperthermia. In the patients with arterio-portal shunt, therapeutic effects were observed in 17% (1/6) and 50% (1/2) of Group A and Group B, respectively. In the patients with portal invasion (Vp2 and Vp3), no favorable response was demonstrated in Group A (0/4), but tumor response was demonstrated in one out of 3 cases in Group B. One- and two-year survival rates were 59% and 53%, respectively, in Group A, and 93% and 45%, respectively, in Group B. The DSM, temporary embolus, is suitable for the combination of hyperthermia because chemo-embolization using DSM can be performed many times. Therefore, these results suggested that chemo-embolization using DSM should be beneficial as combination therapy for unresectable hepatocellular carcinoma. PMID- 2551249 TI - [Carcinogenesis in familial polyposis coli]. AB - The inherited cancer disease familial polyposis coli (FPC) provides an excellent model not only for studying tumor progression in colorectal cancer but also for elucidating molecular mechanism of carcinogenesis in general. This paper reviews recent remarkable progress in the molecular mechanism of tumorigenesis in FPC. Included in this review is information on the localization of the FPC major gene and several types of genetic alterations during the development of tumors. One of these genetic alterations is activation of oncogenes such as mutated ras genes and another is inactivation of tumor suppression genes such as loss of heterozygosity. The mutated FPC gene on chromosome 5 q would be expected to be an early event to initiate the requisite hyperproliferation for adenoma formation. Adenomas will have undergone several gene or chromosome mutations before reaching the fully malignant state. PMID- 2551250 TI - [A randomized phase II study of (2''R)-4'-0-tetrahydropyranyladriamycin and adriamycin in combination with vincristine and ACNU in small cell lung cancer- THP-ADM, VCR, ACNU vs ADM, VCR, ACNU]. AB - Between April 1984 and March 1988, a comparative randomized phase II study was performed to compare the effects of (2''R)-4'-0-Tetrahydropyranyl-adriamycin (THP) and adriamycin in combination with vincristine (VCR) and ACNU in 60 previously untreated and evaluable patients with small cell lung cancer (SCLC). Arm AVA was constituted by adriamycin, VCR and ACNU, and arm TAVA by THP, VCR, ACNU. Of the 30 patients treated with AVA, there were 20 partial responses, 7 with no change and 3 with progressive disease, for an overall response rate of 66.7%. On the other hand, of the 30 patients on TAVA, one complete response and 22 partial responses were observed, for an overall response rate of 76.7% Median survival time of AVA was 10.0 M, that, of TAVA was 9.3 M. But significant differences between the two arms was not found. During induction therapy, leukopenia was the main side effect. Over WHO Grade 3 leukopenia was seen in 53.3% of patients on AVA and 70.0% of those on TAVA. Moderate hair loss (Grade 2) was significantly less frequent with TAVA than AVA. In conclusion, the results indicated that THP is active in SCLC with the same level of adriamycin, and has less toxicity. THP is a suitable drug as a first line combination chemotherapy for SCLC. PMID- 2551252 TI - [Development of a slow release system of anti-cancer drugs retained in calcium hydroxyapatite ceramic]. AB - A new type of anticancer material with sustained release by enclosure of cisplatin (CDDP) into porous calcium hydroxyapatite ceramic (CDDP-CHA) was developed. A slow release of anticancer drug from CDDP-CHA was confirmed in in vitro experiments. When this material was implanted into normal thigh muscle of mouse, a sustained release of CDDP was observed during over 8 weeks after implantation. The diffusion of drug into blood and other organs from the implanted site was very small. CDDP-CHA placed into the implanted tumor of mouse allowed a slow release and high drug level from the material in the tumor. The concentration of the drug in other organs such as liver and kidney was very low compared with that of the tumor. These results suggest that this material has an important role for cancer chemotherapy, particularly in bone tumors for the reasons of calcium hydroxyapatite ceramic carrier having a mechanical strength. PMID- 2551251 TI - [Study on the pharmacokinetics of UFT in patients with hepatocellular carcinoma associated with cirrhosis]. AB - UFT was administered preoperatively in 20 cases of primary hepatocellular carcinoma, and tegafur, 5-fluorouracil (5-FU), uracil, total thymidylate synthase (TS) and free TS in blood and liver tissue were determined. The results were as follows. 1. Tegafur level was significantly high in blood, while the levels of 5 FU and uracil were high in the liver tissue. The 5-FU level in the cancerous area was 0.099 micrograms/g, higher than the effective level regardless of the presence or not of complication with liver cirrhosis. 2. Total TS, FdUMP (computed as total TS-free TS) and TS inhibition rate [(computed as (TS-free TS)/TS x 100 (%)] were significantly higher in the cancerous liver tissue than in the non-cancerous area. In the cases where determination was made simultaneously for the cancerous liver and non-cancerous liver tissues, the FdUMP level and TS inhibition rate in the cancerous liver tissue were high in 14 out of 17 cases (82.4%) and 13 out of 17 cases (76.5%) respectively. Therefore, UFT seems to have selective toxicity. 3. No correlation was found between 5-FU level, total TS, FdUMP in the liver tissue on the one hand and TS inhibition rate on the other. 4. UFT yielded sufficient 5-FU levels in the cancerous area regardless of the presence or not of complication with cirrhosis and therefore is expected to inhibit DNA synthesis selectively in the cancerous tissue. PMID- 2551253 TI - [A case of hepatocellular carcinoma effectively treated by intraarterial infusion of CDDP and other agents]. AB - It is very common for intraarterial infusion therapy of some anticancer agent to be effective against hepatocellular carcinoma. In this case, the patient was a 74 year-old man who suffered from very advanced hepatocellular carcinoma with tumor thrombus of the intrahepatic portal vein and IVC. He was treated with intraarterial infusion of CDDP, Etoposide, 5-FU, through a catheter placed in the proper hepatic artery. CDDP (30 mg/day) and Etoposide (60 mg/day) were given once every 5 days, and then 5-FU(250 mg/day) was infused daily for 26 days. The patient underwent this protocol study twice in 3 months. After the intraarterial infusion, transarterial embolization using CDDP (100 mg) powder added to lipiodol and aluminum stearate as suspension was done a month later. The tumor regression rate was 84% after intraarterial infusion of CDDP, Etoposide and 5-FU. The tumor thrombus in the intrahepatic portal vein and IVC had completely disappeared. We could not find lipiodol accumulated in the tumor after TAE. Thus, we assumed that the remaining tumor was a necrotic scar and that a complete response was obtained in the patient. There were some side effects, such as nausea, vomiting, pancytopenia and gastritis but no severe complication occurred. PMID- 2551254 TI - [NK activity and T cell subsets in percutaneous ethanol injection therapy of liver cancer--effect of lentinan with combined use]. PMID- 2551255 TI - [Effect of cisplatinum (CDDP) and combined chemotherapy with CDDP on peripheral blood lymphocyte natural killer (NK) activity]. PMID- 2551256 TI - [Non-vascular interventional radiology--percutaneous ethanol injection (PEI) in hepatocellular carcinoma smaller than 3 cm in diameter]. AB - Percutaneous ethanol injection (PEI) was carried out for 109 lesions in 85 patients with hepatocellular carcinoma (HCC) smaller than 3 cm in diameter during the period from Dec. 1982 to Mar. 1989. Therapeutic effect of PEI on HCC was evaluated by ultrasound showing that 15 of 68 HCCs disappeared and the remaining 53 HCCs decreased in size with regression rates ranging from 8% to 88% at 6 months after PEI. Survival rates from PEI calculated by the Kaplan-Meier method showed a 1-yr survival of 92%, a 2-yr survival of 81%, a 3-yr survival of 61%, a 4-yr survival of 55%, and a 5-yr survival of 41%. These survival rates were better than those of patients with HCCs smaller than 3 cm in size who had not received anti-cancer treatment (p less than 0.05). PMID- 2551257 TI - Upper limb pyrophosphate tenosynovitis outside the carpal tunnel. AB - Three cases of calcium pyrophosphate dihydrate (CPPD) crystal deposits in tendon sheaths outside the carpal tunnel are reported. Crystals were shown by x ray diffraction analysis in one case and by compensated light microscopy in the other two. Surgical excision of the tendon synovial sheath had to be done in two cases (one case with CPPD crystal deposits). PMID- 2551258 TI - Activation and distribution of rat parotid cAMP-dependent protein kinase following beta-adrenergic receptor stimulation in vitro. AB - The extent of activation of parotid protein kinase A (EC 2.7.1.37) isozymes was determined using dispersed cells and an 8-N3-[32P]-cAMP photoprobe. Cold-trap studies indicated that 40% of type I protein kinase A was activated following maximal beta-adrenergic receptor stimulation, whereas type II activation was less than 20%. Both cytosolic and microsomal type I activation occurred rapidly after stimulation and both remain activated throughout the entire secretory period. The dose-response relationship for the isotypes following beta-adrenergic receptor activation demonstrated a greater extent of type I activation at maximal concentration of agonist. Although protein kinase A may not be the only kinase involved in rat parotid amylase release, these findings add further evidence of a direct regulatory role for this kinase, with type I having potentially a greater role than type II. PMID- 2551259 TI - Rehabilitation for stroke survivors--a review. PMID- 2551260 TI - Prevalence of colon diverticulosis in Europe. PMID- 2551261 TI - Dietary treatment with fibre in large bowel disease. PMID- 2551262 TI - Nutritional prevention of diseases of the large intestine. PMID- 2551263 TI - The role of low-calorie foods in weight reduction. Opening of the discussion. PMID- 2551264 TI - Renal necrosis and the involvement of a single enzyme of the de novo pathway for the biosynthesis of platelet-activating factor in the rat kidney inner medulla. AB - 2-Bromoethylamine hydrobromide (BEA), when administered to rats, induces a highly specific papillary necrosis associated with the inner medulla. PAF levels in the blood were lowered by 50% and of the three enzymes that comprise the de novo route for PAF in the cortex/medulla, only the cholinephosphotransferase activity in the inner medulla microsomes was reduced (33%) by the BEA treatment. Moreover, BEA did not affect phosphatidylcholine synthesis in either the cortex or inner medulla. Our studies indicate that the de novo pathway for PAF synthesis in the renal inner medulla is responsible for the secretion of newly formed PAF into the blood stream and that a single enzyme in the de novo route accounts for the decreased rate of PAF synthesis during the development of renal necrosis. PMID- 2551265 TI - Indirect evidence that protein kinase C plays a critical role in signal transduction of both vasopressin and corticotropin-releasing factor on pituitary cells in culture. AB - The possible role of protein kinase C (PKC) in the cyclic AMP-dependent mechanism of action of corticotropin-releasing factor (CRF) on proopiomelanocortin cells of anterior and intermediate pituitary glands was examined after pretreatment of cells in culture with the PKC inhibitor retinal or the phorbol ester PMA, which depletes cell stores of the kinase. We found that these drugs not only abolished ACTH response to PMA and vasopressin, which both activate PKC, but unexpectably also dampened by 80-90% the stimulatory effect of CRF. Cell treatment with retinal failed to prevent CRF-induced accumulation of cyclic AMP. Retinal and PMA pretreatments of intermediate pituitary cells likewise inhibited alpha-MSH secretion stimulated by CRF. These data provide evidence to suggest that the mechanism of action of CRF on pituitary cells involves both cyclic AMP and PKC messenger systems. PMID- 2551266 TI - Tumor necrosis factor-alpha inhibits follicle-stimulating hormone-induced differentiation in cultured rat granulosa cells. AB - We have investigated the effects of TNF-alpha on FSH-induced LH receptor expression, cAMP and progesterone production in cultured rat granulosa cells. TNF alpha (0.5-100 ng/ml) inhibits the stimulating action of FSH on LH receptor formation in a dose-dependent manner with an IC50 of 1 ng/ml and an almost complete suppression of LH receptor induction for 50-100 ng/ml TNF-alpha. The inhibitory effect of TNF-alpha is not due to variations in cell number or viability but rather to a reduction of the LH receptor content per cell with no change in binding affinity (KD = 0.8 x 10(-10)M). TNF-alpha also inhibits the FSH induced cAMP production but at a lower extent, with a maximum reduction of 60% for 100 ng/ml TNF-alpha. Moreover, TNF-alpha impairs the LH receptor formation induced by forskolin, cholera toxin or 8-Bromo-cAMP, indicating that the cytokine also acts at a step distal to FSH receptor and to cAMP formation. Finally, TNF alpha decreases dramatically the progesterone synthesis that is stimulated by FSH, with a reduction to undetectable levels on and after 10 ng/ml TNF-alpha. These results suggest that TNF-alpha may drastically reduce the capacity of granulosa cells to differentiate upon FSH stimulation and to respond to LH during the physiological ovarian follicular maturation. Such anti-gonadotropic action of TNF-alpha on granulosa cell differentiation may be also relevant to the alteration of ovarian function during physiopathological processes like inflammatory or infection diseases. PMID- 2551267 TI - Evidence for a novel metabolic pathway of (ADP-ribose)n: pyrophosphorolysis of ADP-ribose in HeLa S3 cell nuclei. AB - ADP-ribose liberated from (ADP-ribose)n by the action of (ADP-ribose)n glycohydrolase was converted to ATP and ribose 5-phosphate (ribose 5-P) in the presence of pyrophosphate (PPi) in HeLa S3 cell nuclei. This reaction was reversible and dependent on the simultaneous presence of ADP-ribose, PPi, Mg2+, and nuclei. These results suggest the presence of a novel enzyme in the nuclei, designated as ADP-ribose pyrophosphorylase, which catalyzes the reaction shown in Equation 1. ADP-ribose + PPi in equilibrium ATP + Ribose 5-P (1) This reaction could represent a pathway for the biosynthesis of ATP from (ADP-ribose)n in eukaryotic cell nuclei. PMID- 2551268 TI - Continuous spectrophotometric assay for retroviral proteases of HIV-1 and AMV. AB - Ac-Lys-Ala-Ser-Gln-Asn-Phe(NO2)-Pro-Val-Val-NH2 (peptide I) and Thr-Phe-Gln-Ala Phe(NO2)-Pro-Leu-Arg-Glu-Ala (peptide II) undergo hydrolysis between the p nitrophenylalanyl and prolyl residues catalyzed by the proteases of HIV-1 and AMV, respectively. The specific hydrolyses of peptides I and II are accompanied by a decrease in their uv absorption at 269 nm (delta epsilon = 1000) and an increase at 316 nm (delta epsilon = 600). The use of microspectrophotometric cells allows continuous uv measurements on a volume (60 to 120 microliters) comparable to that required for the HPLC point assay currently used. At the highest substrate concentration possible under the assay conditions, good first order kinetics were observed with both proteases, and the values of Vmax/Km were obtained. PMID- 2551269 TI - ADP and, indirectly, ATP are potent inhibitors of cAMP production in intact isoproterenol-stimulated C6 glioma cells. AB - When added to intact C6 glioma cells in the micromolar range of concentrations, ADP and ATP induce an inhibition of the isoproterenol-elicited cAMP responses. ATP is rapidly hydrolyzed by the ectonucleotidases present on these cells, with an apparent Km of 50 microM and a Vmax of 1.1 nmol/min/10(6) cells. cAMP responses are also inhibited by millimolar concentrations of either ATP in the presence of an ATP-regenerating system to prevent ADP accumulation or AMP-PCP. These observations show that, in C6 glioma cells, ADP is a more potent inhibitor of cAMP production than ATP, the latter acting indirectly, via its rapid hydrolysis to ADP. The additive inhibition of isoproterenol-elicited cAMP responses induced, on one hand, by the treatment of the cells with a phorbol ester and by addition of ADP to the cells, and, on the other hand, by the progressive disappearance of the effects of ADP and ATP when cells are treated with increasing concentrations of Pertussis toxin, demonstrate that ADP and ATP exert their action in C6 glioma cells via a P2 purinoceptor probably negatively coupled to adenylate cyclase and a G regulatory protein. PMID- 2551270 TI - Identification of a dihydropyridine-sensitive calcium channel in chick brain by a monoclonal antibody. AB - A Balb/c mouse was immunized with chick synaptic plasma membranes and monoclonal antibodies were produced by fusion of spleen cells with NS-1 mouse myeloma cells. One antibody, MAC-L1, immunoprecipitated more than 90% of the [3H]PN200-110 labeled calcium channels but only 20% of the omega -conotoxin receptor solubilized from the chick brain membranes. Thus possibly, a certain portion of the omega -conotoxin receptor in the chick brain is a dihydropyridine-sensitive calcium channel. By the specific immunoprecipitation of 125I-labeled proteins, two large polypeptides of 193kDa and 130kDa under reducing conditions were identified as the major components of the calcium channel. PMID- 2551271 TI - Pancreastatin: a novel peptide inhibitor of parietal cell signal transduction. AB - The direct inhibition of secretion by pancreastatin was investigated in rabbit isolated parietal cells. Pancreastatin exerted no influence on basal aminopyrine uptake. Pancreastatin inhibited histamine stimulated aminopyrine uptake through a decrease in intracellular cAMP. Pancreastatin inhibition of histamine stimulated uptake was blocked in the presence of pertussis toxin. Pancreastatin also inhibited the carbachol stimulated increase in aminopyrine accumulation. However, the effects of pancreastatin on carbachol stimulation were not reversed by pertussis toxin. Pancreastatin did not alter the carbachol induced increase in cytosolic free calcium. Thus, pancreastatin appears to inhibit parietal cell signal transduction at multiple points along the second messenger pathways. PMID- 2551272 TI - Multiple defects of the respiratory chain including complex II in a family with myopathy and encephalopathy. AB - We report severe deficiency of complex II of the mitochondrial respiratory chain and low activities of complex I and II in skeletal muscle mitochondria from a young woman with progressive muscle weakness and encephalopathy. Defects of complex II have only very rarely been described and this is the first report of decreased immunoreactive subunits associated with severe deficiency of this enzyme. PMID- 2551273 TI - Evidence for a single active site in the human angiotensin I-converting enzyme from inhibitor binding studies with [3H] RU 44 403: role of chloride. AB - Binding of the potent radiolabelled competitive inhibitor 3H RU 44403 to pure human kidney angiotensin-I converting enzyme was examined in equilibrium and non equilibrium conditions. Equilibrium dialysis experiments indicate that, despite the duplicated structure of the enzyme and the presence of two putative active sites, 3H RU 44403 interacts with a single high affinity (Kd = 0.44 +/- 0.05 x 10 9 M, n = 3) binding site. This suggests that only one of the two putative active sites is functional, and can bind substrates or inhibitors. Sodium chloride plays an essential role in the enzyme-inhibitor interaction. The formation of the complex is only slightly influenced by NaCl, but the kinetic of dissociation is dramatically dependent on NaCl concentration. In a Nacl free medium the complex is unstable and dissociates rapidly. These results are consistent with the hypothesis that chloride ion influences isomerization of the complex toward a more stable form. PMID- 2551274 TI - Differential contribution of C-terminal regions of dermorphin and dermenkephalin to opioid-sites selection and binding potency. AB - Dermorphin and dermenkephalin are D-aminoacid containing peptides generated from processing of the plurifonctional biosynthetic precursor pro-dermorphin. Dermorphin, Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH2 (DRM) and dermenkephalin, Tyr-D-Met Phe-His-Leu-Met-Asp-NH2 (DREK), are among the most selective and potent agonists described respectively for the mu- and delta-opioid receptors. In order to identify determinants of selectivity and high-affinity receptor binding of dermorphin and dermenkephalin, a series of analogs was investigated for their affinity at the mu- and delta-receptors in the brain. The tetrapeptide amino end of both DRM and DREK were found to display high affinity and selectivity towards mu-receptors. Substitution of the C-terminal tripeptide of DREK with that of DRM reversed the receptor selectivity of DREK from delta to mu. Replacement of the C terminal tripeptide of DRM with the C-terminal counterpart of DREK shifted the selectivity of DRM from mu to delta. These data emphasize the critical contribution of the carboxy end of DREK to delta-selectivity. They further suggest that the potent mu-address lying in the N terminus of DREK is overwhelmed by the powerful delta-directing ability of the carboxy end. Unlike DREK, the C terminus of DRM is not involved in opioid receptor sites selection but is important insofar as it serves to stabilize interactions of DRM with the mu receptor binding site. PMID- 2551275 TI - Acid-promoted tautomeric lactonization and oxidation-reduction of pyrroloquinoline quinone (PQQ). AB - Acid-treatment facilitates PQQ detection by electron ionization mass spectroscopy with a molecular ion at M/e 330 and a base ion formed by triple decarboxylation at M/e 198. Other ions found probably arise through acid-catalyzed tautomeric lactonization of PQQ to PQQ-lactone (PQQL) with subsequent oxidation of PQQL and reduction of PQQ. We propose that a carboxyl group, presumably the 9-carboxyl, attacks a double bond in PQQ, reversibly converting the 4,5-orthoquinone into an 4,5-enediol and forming an isomeric lactone, PQQL, of 330 daltons. The masking of carbonyls may explain the low reactivity of PQQ with carbonyl reagents in acid. Acid-promoted tautomeric lactonization with carbonyl-masking is known to occur with fluoresceins, phenolphthalein and other compounds, but has not been recognized before with PQQ. Acid-treated PQQ demonstrates molecular and other ions derived from reduced PQQ (PQQ(2H] or its lactone at M/e 332 with a base ion at M/e 200. There is compelling evidence for a dehydrogenated lactone, PQQ( 2H)L), at M/e 328 with a base ion at M/e 196. We suggest that PQQ, in tautomeric equilibrium with PQQL, oxidizes PQQL to PQQ(-2H)L (328 daltons), with its concurrent reduction to PQQ(2H) (332 daltons). With acidified D2O, PQQ shows deuterated products with ions at M/e values consistent with lactonization and oxidation-reduction. An analytically useful quinoxaline adduct, formed from PQQ and 2,3-diaminonaphthalene (PQQ-DAN) of 452 daltons, also undergoes acid tautomerization-lactonization and oxidation-reduction similar to PQQ showing molecular ions at M/e 450, 452 and 454 and decarboxylation-derived strong (base) ions at M/e 318, 320 and 322.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551276 TI - Angiogenin depresses aortic smooth muscle cell cAMP by a pertussis toxin sensitive mechanism. AB - Angiogenin transiently depresses the cAMP level of rat aortic smooth muscle cells. The dose response is similar to angiogenin activation of the inositol specific phospholipase C in this cell line [Moore, F. & Riordan, J.F. (1989) Biochemistry. Submitted]. The time course showed a maximal depression (28%) in cAMP at 2 min, followed by a return to that of unstimulated cells by 3.5 min. Angiogenin also inhibited isoproterenol stimulated cAMP formation, but the percentage depression in cAMP (9%) was less than that in cells treated with angiogenin alone (28%). In contrast angiogenin enhanced forskolin stimulation of adenylate cyclase, an effect previously linked with agonist activation of protein kinase C. The effect of angiogenin on cellular cAMP was abolished by pre incubation with pertussis toxin. Angiogenin had no effect on cellular cGMP. These results are consistent with activation of adenylate cyclase Gi following exposure of the cells to angiogenin and provide further evidence for interaction between cellular signalling pathways. PMID- 2551277 TI - Direct regulation of smooth muscle contractile elements by second messengers. AB - The effects of adenosine 3',5'-cyclic monophosphate (cAMP), guanosine 3',5' cyclic monophosphate (cGMP) and phorbol 12,13 dibutyrate (PDBu) on the Ca2+ sensitivity of the contractile elements in the rat mesenteric artery were investigated, using a method of permeabilizing smooth muscle with Staphylococcal alpha-toxin. Both cAMP and cGMP relaxed the permeabilized rat mesenteric artery at the intracellular Ca2+ concentrations [( Ca2+]i) held constant with Ca2+ EGTA buffer and Ca2+ ionophore, ionomycin. In addition, forskolin and sodium nitroprusside which activate adenylate and guanylate cyclases, respectively, also induced relaxation at a fixed [Ca2+]i. In contrast PDBu which stimulates protein kinase C caused an increase in force at a constant [Ca2+]i which could be partially reversed by cAMP or cGMP. These results indicate that second messengers exert direct control over smooth muscle Ca2+ sensitivity of the contractile elements, which is of physiologic and pharmacologic importance. PMID- 2551279 TI - Binding of insulin-like growth factor-I by glomerular endothelial and epithelial cells: further evidence for IGF-I action in the renal glomerulus. AB - The renal glomerulus is both a site of action and synthesis of IGF-I. We previously demonstrated the presence of IGF-I receptor and synthesis in glomerular mesangial cells. In this study we investigated the presence of specific IGF-I receptors on mouse glomerular endothelial and epithelial cells in culture. [125I]IGF-I specifically bound to the cell surface of both cell types. Maximum specific binding, 0.141 B/F for endothelial cells and 0.301 B/F for epithelial cells, was obtained at 22 degrees C after 150 min incubation. The estimated Kd values were 2.25 x 10(-9) for endothelial cells and 1.5 x 10(-9) for epithelial cells. Cross-linking studies showed a single band of radioactivity with an estimated mol wt of 145K, consistent with the alpha-subunit of the IGF-I receptor. Radiolabelled IGF-I was not degraded by either cell types. These findings suggest a possible paracrine action of IGF-I in the renal glomerulus. PMID- 2551278 TI - Functional endothelin/sarafotoxin receptors in rat heart myocytes: structure activity relationships and receptor subtypes. AB - Functional receptors for the peptides of the endothelin (ET) and sarafotoxin (SRTX) family were characterized in newborn rat heart myocytes using human and rat endothelins (ET-1 and ET-3, respectively), SRTX-b and SRTX-c. Binding studies in intact cells and homogenates revealed significantly higher affinities of ET-1 and SRTX-b than of ET-3 and SRTX-c towards these receptors. This binding profile of ET/SRTX peptides points to their interaction with the receptor subtype designated E-S alpha. All four peptides induced time- and dose-dependent phosphoinositide hydrolysis with the following rank order of potency: ET-1 greater than SRTX-b greater than SRTX-c greater than ET-3. Thus, ET-3 which possesses an intermediate affinity toward the receptor was the least effective with regard to this response. These results confirm and extend our earlier report that the ET/SRTX peptides interact with a newly characterized receptor(s) associated with phosphoinositide metabolism and Ca2+ mobilization. The initiation of inositol phosphate formation is largely independent of extracellular Ca2+, verapamil and nifedipine, indicating that the ET/SRTX peptides are not agonists for the voltage-dependent Ca2+-channels. PMID- 2551280 TI - Stimulation of protein phosphorylation by atrial natriuretic factor in plasma membranes of bovine adrenal cortical cells. AB - Atrial natriuretic factor (ANF) rapidly enhanced phosphorylation of plasma membrane proteins of bovine adrenal cortical cells. Pretreatment of the membranes with ANF (1 x 10(-8)M to 1 x 10(-7)M) resulted two- to four-fold in an incorporation of 32p-radioactivity from [gamma -32p]ATP as compared to the controls. The guanosine 3', 5' monophosphate (cGMP) which has been considered a second messenger of ANF also enhanced the phosphorylation of several proteins which were stimulated by ANF. However, the phosphorylation of certain proteins was stimulated differentially only by either ANF or cGMP. These results suggest that ANF-induced protein phosphorylation may play a role in transmembrane signalling pathway involving other second messenger(s) in addition to cGMP during the biological action of ANF. PMID- 2551281 TI - The identification of a novel inositol lipid, phosphatidylinositol trisphosphate (PIP3), in rat cerebrum using in vivo techniques. AB - Rats received intraventricular injections of 20 uCi of [3H]-myo-inositol, and were sacrificed 24 hrs later by high-power head-focused microwave fixation. Two inositol lipid extraction methods were compared: The Hauser and Eichberg method yielded higher recovery of inositol lipids, but a lower inositol phosphate content. The Schacht method yielded reduced radiolabel in the lipid fractions, but increased water soluble phosphates. Both methods extracted a novel inositol lipid (PIP3) which contained inositol tetrakisphosphate (IP4) as its polar head group. This was determined by alkaline hydrolysis and analyzed by high performance liquid chromatography with authentic IP4 standard. Furthermore, preliminary studies of the fatty acid composition indicated a similarity with other inositol lipids. The radiolabel ratio of PIP2:PIP3 was 5:1. In summary, we have isolated a novel inositol phospholipid in rat brain, PIP3, the parent compound for inositol tetrakisphosphate (IP4). PMID- 2551282 TI - Direct activation by prostaglandin F2 alpha but not thromboxane A2 of glycogenolysis via an increase in inositol 1,4,5-trisphosphate in rat hepatocytes. AB - In rat liver prostaglandin F2 alpha (PGF2 alpha) and thromboxane A2 (TXA2), released from non-parenchymal cells, have been implicated as mediators of the enhancement of glucose and lactate output from parenchymal cells caused by sympathetic nerve stimulation [Iwai, M. et al. (1988) Eur. J. Biochem. 175, 45 50]. In isolated rat hepatocytes PGF2 alpha, of which 75% were degraded within 10 min, but not the TXA2 analogue U46619 increased inositol 1,4,5-trisphosphate (IP3), glycogen phosphorylase a activity and glucose output like noradrenaline and vasopressin; cyclic AMP remained unaltered. The maximal increase in IP3 was reached within 20 s and in phosphorylase activity as well as glucose release within 1 min. The results indicate that only PGF2 alpha but not TXA2 can play a role as a direct mediator of the sympathetic metabolic nerve actions in rat liver and that hepatocytes contain also stimulatory prostaglandin receptors linked to phospholipase C in addition to the inhibitory receptors linked to adenylate cyclase known thus far. PMID- 2551283 TI - High-efficiency expression of mammalian beta-adrenergic receptors in baculovirus infected insect cells. AB - Infection of a clonal isolate of Spodoptera frugiperda cells (Sf9) with a baculovirus expression vector harboring the cDNA encoding the beta-adrenergic receptor resulted in a high efficiency expression. At 48 hr post-infection, the level of expression of beta-adrenergic receptors was approximately 12 million/cell. Specific activities of crude lysates of infected Sf9 cells were approximately 30 pmol/mg of protein, 5-fold greater than those of membranes of high-expressor Chinese hamster ovary cells stably transfected with an SV-40 expression vector. One liter of infected Sf9 cells expresses 20-40 nmol of receptor. Autoradiography of membranes incubated with the beta-adrenergic antagonist [125I]iodoazidobenzylpindolol, photolyzed, and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed 46,000- (presumably unglycosylated) and 48,000-Mr peptides for Sf9 cells as compared to approximately 65,000-Mr for Chinese hamster ovary cells. The baculovirus Sf9 system provides high-efficiency expression of receptor sufficient to permit physicochemical analyses. PMID- 2551285 TI - Stimulation of phosphoinositide signaling pathway in murine B lymphocytes by a novel guanosine analog, 7-thia-8-oxoguanosine. AB - A novel guanosine analog, 7-thia-8-oxoguanosine, is shown to induce proliferation in nude mouse splenic lymphocytes in vitro as measured by increased DNA synthesis. Determination of the levels of sn-1,2-diacylglycerol and inositol 1,4,5-trisphosphate in the 7-thia-8-oxoguanosine-treated B lymphocyte cultures revealed an enhanced formation of these second messengers leading to the activation and possible de novo synthesis of protein kinase C. The relevance of these results are discussed in regard to the transduction mechanism linking these guanosine analog generated signals with subsequent B lymphocyte activation events. PMID- 2551284 TI - Cyclosporine increases endothelin-1 binding site density in cardiac cell membranes. AB - When used in high concentrations, the immunosuppressive agent cyclosporine A has toxic side effects, including cardiac fibrosis and calcification. Cyclosporine treated mice were used to investigate whether a cyclosporine dose regime which produces cardiac fibrosis and calcification alters the density of the specific binding sites for the endothelial-derived vasoconstrictor polypeptide, endothelin 1. After twenty-one days of treatment the endothelin-1 binding site density in cardiac cell membranes was increased, without any change in selectivity. This increase in endothelin-1 binding site density could contribute to the cytotoxic effects of cyclosporine. PMID- 2551286 TI - Synergistic deleterious effect of micromolar Ca ions and free radicals on respiratory function of heart mitochondria at cytochrome C and its salvage trial. AB - Both Ca2+ and free radicals (FR) are accumulated in temporarily ischemic myocardium and might cause reperfusion injury. Respiratory function measured by polarography of isolated heart mitochondria before or after the in vitro treatment with Ca2+ (1.2 microM) and/or FR showed that Ca2+ or FR per se showed no or weak effect on state 3, but cotreatment of Ca2+ and FR prominently deteriorated state 3, state 4 and RCI. The injury was speculated to occur at cytochrome c itself or its reductase from enzyme assay in the respiratory chain. Furthermore, contrary to the published data, the synergistic action was not mitigated by phospholipase A2 inhibitors (dibucaine, mepacrine), membrane stabilizers (lidocaine, coenzyme Q10), Ca entry blocker (verapamil) or superoxide dismutase, suggesting refractory to therapy. PMID- 2551287 TI - Identification of a major GTP-binding protein in bovine aortic smooth muscle membranes as smg p21, a GTP-binding protein having the same effector domain as ras p21s. AB - At least two GTP-binding proteins (G proteins) with Mr values of about 20,000 were extracted from bovine aortic smooth muscle membranes by sodium cholate. The most abundant G protein (22K G) was purified to near homogeneity by successive column chromatographies of Ultrogel AcA-44, phenyl-Sepharose CL-4B, hydroxyapatite and Mono Q HR5/5. 22K G showed kinetic and physical properties very similar to those of smg p21, a G protein recently isolated from bovine brain and human platelet membranes, having the same effector domain as ras p21s. Moreover, 22K G was recognized specifically by the anti-smg p21 antibody. These results indicate that the major G protein in bovine aortic smooth muscle membranes is smg p21. PMID- 2551288 TI - Action of atrial natriuretic peptide and angiotensin II on the myocardium: studies in isolated rat ventricular cardiomyocytes. AB - Isolated calcium-tolerant rat ventricular cardiomyocytes were used to characterize the effects of atrial natriuretic peptide (ANP), Angiotensin II (AII) and their interaction on the myocardial contraction-/relaxation pattern free of interference from other types of cardiac cells. Binding of 125I-ANP showed a KD of 12 pM and approximately 600 binding sites per cell. At 37 degrees C (rate 140 bpm) ANP decreased the contraction maximum with an EC50 of about 70 pM, maximal decrease was 35%. ANP (10(-7) M) raised cellular cyclic-GMP from 0.76+/-0.12 to 1.32+/-0.13 pmole/10(6) cells (73%, p less than 0.05). Angiotensin II increased contractility by a maximum of 32% at 10(-7) M; the EC50 was 8 x 10( 10) M. AII markedly delayed relaxation (reduction of maximum relaxation velocity from 0.092 to 0.063 mm/s; p less than 0.05). ANP (10(-7) M) increased the effect of AII (10(-8) M) on contractility by 66% without changing relaxation parameters significantly. This unexpected interaction may be relevant in pathological conditions where both AII and ANP are stimulated, such as heart failure or secondary hypertension. PMID- 2551290 TI - Deficiencies in complex I subunits of the respiratory chain in Parkinson's disease. AB - Immunoblotting studies on mitochondria prepared from the striata of patients who died of Parkinson's disease were performed using specific antisera against Complexes I, III and IV. In 4 out of 5 patients with Parkinson's disease, the 30 , 25- and 24-kDa subunits of Complex I were moderately to markedly decreased. No clear difference was noted in immunoblotting studies on subunits of Complexes III and IV between the control and Parkinson's disease. Deficiencies in Complex I subunits seem to be one of the most important clues to elucidate pathogenesis of Parkinson's disease. PMID- 2551289 TI - Regulatory activities of 2 beta-(3-hydroxypropoxy)-1 alpha, 25-dihydroxyvitamin D3, a novel synthetic vitamin D3 derivative, on calcium metabolism. AB - Regulatory activities of 2 beta-(3-hydroxypropoxy)-1 alpha, 25-dihydroxyvitamin D3 [ED-71], a novel synthetic vitamin D3 derivative, on calcium metabolism were investigated. The compound behaved similar to 1 alpha, 25-dihydroxyvitamin D3 [1,25(OH)2D3] in the ex vivo intestinal calcium transport using rat everted gut sac and the in vivo bone mobilization using vitamin D-deficient rats. By means of Raisz's assay method, 45Ca releasing activity of ED-71 was not greater than that of 1,25(OH)2D3. The time course curve of ED-71 in plasma made a mild round shape compared with that of 1,25(OH)2D3 and the former's plasma concentration remained increased longer than the latter's. The therapeutic effect of ED-71 for the animal models with osteoporosis seemed to be better than that of 1,25(OH)2D3. The results suggest that ED-71 may be a promising drug for therapy of osteoporosis. PMID- 2551291 TI - Protein kinase C blockers and neutrophil receptors for leukotriene B4. AB - Three protein kinase C blockers (staurosporin, Cl, and sphinganine) acted temperature- and time-dependently on human neutrophils to lower the affinity and number of high affinity plasmalemma receptors available to leukotriene B4. The drugs did not alter the ligand's binding to isolated plasma membranes or reduce intact cell binding of platelet-activating factor. Thus, protein kinase C may regulate the expression of certain receptors in resting cells and blockers of this enzyme, by interfering with receptor expression, have secondary effects that complicate their use as pharmacological probes. PMID- 2551292 TI - Existence of parathyroid hormone binding sites on murine hemopoietic blast cells. AB - We demonstrated that 125I-labeled human parathyroid hormone (1-34;8,18-Nle,34 Tyr)[[125I]hPTH(1-34)] bound specifically to hemopoietic blast cells supported by granulocyte-macrophage colony-stimulating factor. Half-maximal inhibition of binding was achieved at concentrations of unlabeled hPTH(1-34) of about 5 x 10( 9)M. Insulin and hPTH(39-68) did not compete for PTH binding sites. Specific binding of hPTH(1-34) was detected in neither macrophages nor multinucleated cells (MNC's). Furthermore, treatment of hemopoietic blast cells with hPTH(1-34) stimulated MNC formation, and the range of concentrations (10(-10)-10(-8)M) over which hPTH(1-34) caused these effects was similar to that which inhibited the binding of [125I]hPTH(1-34). These findings suggest the presence of a PTH receptor on osteoclast precursors and the direct effect of PTH on them, resulting in osteoclast-mediated bone resorption. PMID- 2551293 TI - The complete primary structure of calcineurin A, a calmodulin binding protein homologous with protein phosphatases 1 and 2A. AB - A complementary DNA (cDNA) clone encoding the catalytic subunit of calcineurin (calcineurin A) has been isolated from a rat brain cDNA library. The primary structure of the cDNA consists of 2,337 nucleotides including the entire coding region for 521 amino acids, and the calculated molecular mass is 58,643 Da. The calcineurin A is strikingly homologous to protein phosphatases 1 and 2A, approximately 50% of the amino acids over an internal 250-residue region between residues 78 and 329 being identical. Twenty four amino acid-residue region between residues 391 and 414 shows the consensus structural features for a calmodulin-binding domain. These data suggest that the allosteric character of this chimeric enzyme is generated by gene fusion of two separate protein families. PMID- 2551294 TI - Molecular cloning of a full-length cDNA for human alpha-N-acetylgalactosaminidase (alpha-galactosidase B). AB - In the process of molecular cloning of cDNA for proteins associated with a purified human placental sialidase fraction, we discovered one of the proteins with apparent molecular weight of 46 kDa is in reality alpha-N acetylgalactosaminidase. The full length cDNA, pcD-HS1204, codes for 358 amino acids with the first 17 residues representing a putative signal peptide. The predicted amino acid sequence shows striking homology with human alpha galactosidase A and yeast alpha-galactosidase. The substrate specificities as well as the behavior of the 46 kDa protein on hydroxylapatite chromatography confirmed that the 46 kDa protein is in reality alpha-N-acetylgalactosaminidase. PMID- 2551295 TI - Proteolysis of tau by calpain. AB - The calpain-induced proteolysis of tau associated with twice-cycled microtubules or from a total brain heat-stable fraction was studied. Twice-cycled microtubule tau was rapidly hydrolyzed by calpain. In contrast, tau purified from the total brain heat-stable fraction was very resistant to degradation by calpain. These results clearly demonstrate that there are at least 2 populations of tau in the brain based on calpain-sensitivity, a calpain-sensitive form that is associated with microtubules and a calpain-resistant form that may represent another population of tau in the brain. PMID- 2551296 TI - Brown adipose tissue (Na+-K+)-ATPase activity and substrate uptake during the breeding cycle of rats. AB - Brown adipose tissue (Na+-K+)-ATPase activity, in vitro glucose and 2 aminoisobutyric acid uptake, as well as mitochondrial GDP-binding and succinate dehydrogenase activity were determined in order to study the relationship between these parameters and the thermogenic status. Analysis were carried out on control animal, pregnant rats, dams and pups during lactation, GDP-binding, (Na+-K+) ATPase and glucose uptake were found to be decreased in brown adipose tissue from pregnant rats and dams, and increased in pups, 2-aminoisobutyric acid uptake was only increased in pups, but no changes were observed in the other experimental groups tested. GDP-binding and (Na+-K+)-ATPase activity showed a parallelism which suggests that the enzyme is a good index of thermogenic status of the animal. PMID- 2551297 TI - Cytosolic adenylate kinase catalyzes the synthesis of thiamin triphosphate from thiamin diphosphate. AB - An attempt was made to purify a porcine skeletal muscle enzyme catalyzing the formation of thiamin triphosphate (TTP) from thiamin diphosphate (TDP), requiring ATP, Mg2+ and a cofactor (creatine). As the purification proceeded, the reaction requirements for ATP and creatine were lost and then a requirement for ADP was manifested. The activity responsible for TTP synthesis from TDP, ADP, and Mg2+ was found to be copurified with adenylate kinase [EC 2.7.4.3] activity, and was finally purified to a single band on SDS-PAGE. Antiserum obtained against the purified enzyme preparation inhibited both adenylate kinase activity and the TTP synthesizing activity to exactly the same extent. These results indicate that adenylate kinase catalyzes TTP formation from TDP in vitro. PMID- 2551298 TI - Properties of the thiamin triphosphate-synthesizing activity catalyzed by adenylate kinase (isoenzyme 1). AB - Adenylate kinase isozyme 1 (AK1) catalyzes thiamin triphosphate (TTP) formation from thiamin diphosphate (TDP) and ADP. The properties of the TTP-synthesizing activity of purified AK1 from porcine skeletal muscle were studied. The activity was found to require TDP, ADP, and Mg2+, and ATP was only 14.4% as active as ADP. Thiamin monophosphate (TMP) and thiamin were not utilized as substrates. ADP was specific as a phosphate donor; and CDP, UDP, and GDP supported TTP formation at rates less than 1% of that with ADP. Optimal pH and temperature for the TTP synthesizing activity were 10.0 and 37 degrees C, respectively. The activity showed saturation kinetics for both substrates, and the Km values for TDP and ADP were calculated to be 0.83 mM and 43 microM, respectively. The enzyme catalyzed the reverse reaction (TTP + AMP----TDP + ADP) and stoichiometry between TTP and TDP was demonstrated in the forward and reverse reactions. PMID- 2551299 TI - The new biology of drug receptors. PMID- 2551300 TI - Effects of the beta 2-adrenoceptor agonist clenbuterol on tyrosine and tryptophan in plasma and brain of the rat. AB - The beta 2-adrenoceptor agonist, clenbuterol (initially 5 mg/kg), was found to significantly reduce plasma tyrosine and raise brain tryptophan levels (P less than 0.01). By comparison, decreases in plasma tryptophan and increases in brain tyrosine were small and often nonsignificant. Amino acid levels measured in different brain regions revealed that the elevations were similar among the cerebellum, striatum, and cortex. These effects were partially blocked by propanolol but not by atenolol. The ED50 was estimated from dose-response curves to be about 0.05 mg/kg for both the decrease in plasma tyrosine and the increase in brain tryptophan. The effects of low doses of clenbuterol were prevented completely by propranolol. Peripheral organs displayed strikingly different patterns of change in amino acid concentrations. Only the spleen had any accumulation of tryptophan, but that was much less than in brain. In contrast, tyrosine and tryptophan were decreased in heart and unaltered in liver; tyrosine was decreased in lung. The elevation in brain tryptophan levels was attenuated by the beta 2-antagonist, ICI 118,551, but not by the beta 1-antagonist, betaxolol; but the reduction in plasma tyrosine was unaffected by either drug. The serotonin antagonist, methysergide, failed to block the effects of clenbuterol. We conclude that changes in amino acid concentrations produced by clenbuterol are mediated by beta 2-adrenoceptor stimulation. Although the increases in brain tyrosine and tryptophan were similar to increases in the plasma ratios of these amino acids to the sum of the other large neutral amino acids competing for transport into the brain, the disparity between the effects of ICI 118,551 in brain and plasma suggests that clenbuterol may also have a direct action in brain to regulate levels of aromatic amino acids. Since clenbuterol has been purported to have an antidepressant effect and since other antidepressants also increase brain tryptophan, this may be a common feature of antidepressant drug action. PMID- 2551301 TI - Detection of free radicals as intermediates in the methemoglobin formation from oxyhemoglobin induced by hydroxylamine. AB - Four distinct paramagnetic intermediates could be observed in the reaction between oxyhemoglobin and hydroxylamine using ESR spectroscopy. The radical species exhibited different stability properties thus different techniques were required for their detection. Two of them were identified as the hydronitroxide radical (NH2O.) and the hemoglobin-nitric oxide complex (Hb2+-NO). The third one is a low-spin iron-(III)-complex, possibly the methemoglobin-hydroxylamine adduct. A fourth paramagnetic species was detected only in the absence of the iron chelator DETAPAC thus indicating that free iron ions were responsible for the formation of this intermediate. The same species was observed when a Fenton system was used to generate the radicals. This species was identified as being the Fe(NO)2X2 complex described in the literature (X = inorganic anions such as OH- or PO3-(4). The identification of the radical intermediates detected in the hydroxylamine-induced methemoglobin formation contributes to a more detailed understanding of the reaction sequence. PMID- 2551302 TI - Inhibition of serum lactate dehydrogenase activity by disulfiram and diethyldithiocarbamate. PMID- 2551303 TI - Interaction of azithromycin and human phagocytic cells. Uptake of the antibiotic and the effect on the survival of ingested bacteria in phagocytes. AB - 14C-labeled azithromycin, a new macrolide antibiotic, was accumulated by various phagocytic cells isolated from volunteers or patients. The concentration of the antibiotic in monocytes, polymorphonuclear leucocytes (PMNLs), and alveolar macrophages was greater than that in the surrounding medium by a factor of between 200 and 668. Azithromycin penetrated somewhat more rapidly into PMNLs and monocytes than into alveolar macrophages. On the other hand the final concentration in the alveolar macrophages was greater by a factor of about 3 than that in the other two phagocytic cells. Staphylococcus aureus, Legionella pneumophila and Haemophilus influenzae previously taken up by the phagocytes were rapidly inactivated by low (0.031-0.5 micrograms/ml) concentrations of the antibiotic, which in the presence of the cells were subinhibitory. There is thus a clear synergism between azithromycin and the phagocytic cells which leads to increased intracellular killing of the bacteria. PMID- 2551304 TI - Effects of new 4-aminopyridine derivatives on neuromuscular transmission and on smooth muscle contractility. AB - The effects of new 4-aminopyridine (4-AP) derivatives were investigated in the isolated mouse phrenic nerve-hemidiaphragm preparation under single impulse stimulation and tetanic conditions. The basic 4-AP structure was modified in position 3 on the pyridine nucleus by introducing different substituents. Results were compared to those obtained with 4-AP and 3,4-diaminopyridine. The compounds were tested for their antagonistic effect against calcium antagonists and botulinum toxin A. The effect on smooth muscle was investigated on the isolated guinea-pig ileum. Physico-chemical parameters of the test compounds were determined by the partition coefficient and ionization constant. Finally structure-activity relationship analysis revealed that the activity was highly related to lipophilicity and the steric volume. So far 4-AP itself provided the most advantageous molecular structure. PMID- 2551305 TI - Effects of lidocaine alone and in combination with tetraethylammonium on the maximum upstroke velocity in guinea-pig papillary muscles. AB - Effects of 18.5 mumol/l lidocaine on the Vmax in the absence and presence of 10 mmol/l tetraethylammonium chloride (TEA), which specifically prolongs the action potential duration with little effects on Vmax, were studied in guinea-pig papillary muscles in a 10 mmol/l K+ Tyrode solution. The frequency-dependent reduction of Vmax was significantly increased at high frequencies (3 and 4 Hz) in the presence of lidocaine plus TEA. A simulation study was performed under the assumption that lidocaine interacts with open, inactivated and resting sodium channels and that TEA specifically prolongs the inactivated state, and a reasonable fit to the experimental Vmax changes was obtained. There were no significant differences between the effects of lidocaine alone and lidocaine plus TEA on the tonic blocks, time constants and zero-time intercepts of reactivation process of Vmax. The F-test shows that these reactivation processes in all preparations examined are described sufficiently well by monoexponential functions, that is, under the assumption of the linear relationship between available sodium conductance and Vmax. The finding that a nonlinear model does not significantly improve the fit indicates the possibility that the nonlinearity may not be prominent under the present experimental conditions. PMID- 2551306 TI - Possible involvement of the dopaminergic system in the mode of action of the potential antidepressant trazium esilate. AB - Trazium esilate (EGYT-3615) is structurally an as-triazino isoquinolinium salt which showed considerable activity in pharmacological tests characteristic for antidepressants (antagonism of tetrabenazine, potentiation of yohimbine, behavioral despair). The compound exhibited minimal sedative effect. Some findings suggest that it influences the central dopaminergic system. The drug potentiated actions of amphetamine such as stereotypy and hypermotility. It differentially blocked the hypothermic and the stereotypy inducing action of apomorphine. Trazium esilate also inhibited the cataleptic state provoked by bulbocapnine in mice. In higher dose it decreased the plasma prolactin level in rats. The compound potentiated the effect of norepinephrine on isolated vas deferens of the rat. Trazium esilate is a weak displacer on a1-, a2- and D2 receptors, however, it induced a2-receptor desenzitization after repeated treatment. It had no influence on rat brain cortical noradrenaline and striatal dopamine release evoked by high K+ concentration, but it increased the spontaneous dopamine outflow in rat striatum. The compound also elevated the striatal dopamine and DOPAC levels both after acute and chronic treatment. PMID- 2551307 TI - New trends in coronary artery disease: the role of granulocyte activation. AB - Circulating granulocytes play an important role in microvascular perfusion and organ pathology. Oxygen radicals released by aggregated and activated neutrophils may exacerbate the tissue damage caused by ischemia. We studied neutrophil aggregation and oxygen metabolites release in 16 patients suffering from coronary artery disease and in 6 control subjects in aorta and coronary sinus blood samples. The neutrophil aggregation (P less than 0.01) and oxidase activity (P less than 0.01) are higher in coronary sinus than aorta samples only in the patients with respect to controls. While the superoxide generation is decreased in coronary sinus (P less than 0.05). Our aggregation assay supported the potential role of granulocytes in neutrophil-endothelial interactions and tissue damage, and our results about oxidase activity and superoxide release suggested the potential role for neutrophil-derived oxygen radicals in the pathogenesis of vascular injury in vivo. PMID- 2551308 TI - [Pleomorphic giant adenomas of the parotid gland]. AB - A 78-year-old lady is described who was admitted to the hospital with a giant mixed tumour of the left parotid gland that had grown to a weight of 4.5 kg within 24 years; among other things a grotesque scoliosis of the entire spine had been developed due to the misload on the spine over many years. The histological examination revealed a type I finding according to Seifert. In this regard, the literature of the last 140 years was reviewed; reports on 31 patients (20 women = 64.5% and 11 men = 35.5%) with giant mixed tumours of the parotid gland between 1 26.5 kg were published during that period. The age of the first tumour manifestation varied between 20 and 40 years. In 3 cases (= 10%) malignant areas were found within the tumour. It appears that at all times erroneous information and action on the part of the family physician has been one of the main reasons for postponing surgical treatment for too long - in fact, for decades. PMID- 2551309 TI - Possible involvement of Epstein-Barr virus (EBV) in polyclonal B-cell activation in Sjogren's syndrome. AB - The following results suggest that EBV might be involved in the mechanism of polyclonal B-cell activation of Sjogren's syndrome (SS). (1) The levels of serum anti-VCA antibodies of both IgG and IgM class were significantly elevated in SS. (2) Excretion of EBV from the oropharynx was frequently observed in SS. (3) Spontaneously transformed B-cell lines producing a large amount of transforming EBV were established preferentially from SS. (4) An EBV-specific regulatory mechanism was impaired in SS. PMID- 2551310 TI - Autostimulatory factor produced by B-cell lines from patients with Sjogren's syndrome. AB - Sjogren's syndrome (SS) is an autoimmune disease with a predisposition to transform into a B-cell lymphoma. Stable B-cell lines were established (without exogenous stimulation other than fetal bovine serum) from the peripheral blood of three SS patients. These cell lines secreted immunoglobulin (either IgG, IgM or both) and expressed cytoplasmic immunoglobulin. They were positive for the B-cell markers Leu 12 and Leu 16, and also for HLA-DR and the transferrin receptor. The cells lacked CD3 and the IL-2 receptor. Supernatants from these cell lines had autostimulatory activity. When 24-h culture supernatants were added to freshly cultured peripheral blood mononuclear cells, the proliferation index was 2-3 times higher as compared to cells cultured with HB101 medium alone. This autostimulatory activity can be attributed to a B-cell growth factor since these supernatants were also able (1) to support the growth of BD9 cells and (2) to augment the proliferation of PB B cells preactivated with S. aureus Cowan strain I. Furthermore, the supernatants did not contain IL-1, IL-2, or gamma-interferon. Thus, B cells that grow spontaneously from the peripheral blood of SS patients spontaneously produce a B-cell growth factor. This factor could contribute importantly to the autoantibody production, tissue lymphoid infiltration and B cell lymphoma seen in this disease. PMID- 2551311 TI - The response to Epstein-Barr virus infection in Sjogren's syndrome. AB - To assess the response to Epstein-Barr virus (EBV) infection in patients with primary Sjogren's syndrome (SS), the frequency of detection of EBV DNA was studied in salivary gland biopsies and the antibody and idiotypic response to the virus was compared with healthy controls and infectious mononucleosis (IM). Viral DNA, detected by in-situ hybridization, was found in biopsies from two out of 12 patients with SS and six out of 10 controls. IgG, IgA and IgM antibodies to the virus, measured by ELISA using synthetic peptides (early antigen and EBNA-1) and a cloned fusion protein (EBNA-1), were normal in sera from 20 patients with SS, whereas infectious mononucleosis patients showed an increase in IgM antibodies to EBNA-1 and IgG antibodies to early antigen. One similarity between infectious mononucleosis and Sjogren's syndrome was a significant increase in the germline heavy chain idiotype G6 in both diseases, suggesting activation of similar B-cell subsets. It is possible that this is due to EBV, though the low frequency of EBV DNA in biopsies and the normal levels of EBV antibodies in SS does not lend any evidence that the virus itself is the causative agent. PMID- 2551312 TI - A novel EBV-related nucleo-cytoplasmic antigen in a null cell-line (HLN-STL-C) reactive to antibodies in the sera from patients with Sjogren's syndrome. AB - We have established a non-T- and non-B-cell line, HLN-STL-C(STL-C), which harbors the EBV genome, from the lymph node cells of a Japanese ATL patient. This cell line expresses a unique Epstein-Barr virus (EBV)-related nucleo-cytoplasmic (N-C) antigen which is detected by indirect immunofluorescence (IF) with the sera from patients with nasopharyngeal cancer (NPC), infectious mononucleosis (IM) or adult T-cell leukemia (ATL). One of the molecular components of this antigen is proved to be STL-C specific 125 kD molecule by immunoblot analysis (IB). To study the involvement of EBV in Sjogren's syndrome (SS), we examined the reactivity of the N-C antigen with the sera of SS patients by IF and IB. Among 24 cases examined, the sera of 21 cases (87.5%) positively stained the N-C antigen by IF. The staining patterns were divided into two types. Type I, (seven cases) showed positive staining for only N-C antigen, and Type II, (14 cases) was positive for N-C antigen associated with diffuse nuclear staining due to antinuclear antibodies in the SS patient's sera. Only one out of 11 non-Sjogren's patients' sera, which were almost all healthy controls, was positive for N-C antigen in this study. By IB, however, only two out of 15 IF-positive SS patients' sera reacted with STL-C specific 125 kD molecule. These results suggested the presence of heterogenous components in the N-C antigen. Our findings may support the hypothetical conception that EBV plays an etiological role in SS. PMID- 2551313 TI - Contemplation of virally induced changes in salivary glands in Sjogren's syndrome: on the use of in situ hybridization in such studies. PMID- 2551314 TI - Cytomegalovirus infection and disease in renal transplant patients treated with cyclosporin. A prospective study. AB - In this prospective study, the incidence of cytomegalovirus (CMV) infection and CMV disease was determined in 175 renal transplant recipients on cyclosporin and low-dose prednisone. CMV infection occurred in 51.4% of the patients, CMV disease in 13.7%. The major manifestations of CMV disease were fever of unknown origin and leukopenia. In the group with CMV infection, there was an increased occurrence of rejection (60% in infected vs 27% in noninfected patients). In most patients (41/54), the rejection preceded the CMV infection. CMV infection did not lead to a decreased graft survival. There was no close time relationship between the onset of clinical symptoms of CMV disease and the laboratory confirmation of CMV infection. A subgroup of patients at risk for the development of severe CMV disease could not be identified. PMID- 2551315 TI - Cytomegalovirus infection in orthotopic liver transplantation. AB - We retrospectively studied 175 orthotopic liver transplants in 151 patients. Of the 151 patients, 59 (39.1%) were diagnosed as having cytomegalovirus (CMV) infection. The rate of infection in patients treated for rejection was 48.8% as compared to 26.2% in patients without rejection (P less than 0.01). Antirejection therapy was associated with culture-positive cases in 33 out of 43 patients as compared to 9 out of 16 patients who had CMV antibody titer elevations. Patients were treated with gancyclovir if they had simultaneous positive cultures from multiple sites and were seriously ill. Eighteen of the 19 patients thus treated had side effects, one of which was serious (bone marrow hypoplasia). Cultures became negative in 15 out of 17 (88%) of the surviving patients. Patient survival was similar to our overall survival rate of 87%. PMID- 2551316 TI - Binding characteristics reveal partial GABA agonist activities of SR 95531. PMID- 2551317 TI - Vasopressin receptor subtypes: autoradiographic localization of V1 vasopressin binding sites in rat brain and kidney. PMID- 2551319 TI - Interaction of alkali ions with Na,K-ATPase. PMID- 2551318 TI - High-affinity choline transporter in synaptosmal membranes. PMID- 2551320 TI - The systems of photo- and pheromone signals transduction in unicellular eukaryotes. PMID- 2551321 TI - Is a water-soluble form of TTX-sensitive channel possible? PMID- 2551322 TI - Synthesis and characterization of conotoxin IIIa. PMID- 2551323 TI - Effects of chemical modification on Na channel function. PMID- 2551324 TI - Interaction of components of the adenylate cyclase system in the course of hormonal signal transmission. PMID- 2551325 TI - Association-dissociation of purified subunits of GTP-binding proteins measured by fluorescence energy transfer. PMID- 2551327 TI - Molecular modeling study of calcium channel activating and blocking 1,4 dihydropyridine derivatives: a hypothetical molecular receptor model. PMID- 2551326 TI - Structural and functional studies of cyclic GMP phosphodiesterase. PMID- 2551328 TI - Characterization of apamin-binding protein associated with a Ca2+ -activated K+ channel. PMID- 2551329 TI - Slow components of the gating current in the frog node of Ranvier. PMID- 2551330 TI - Alpha 2 adrenoceptors of blood platelets from hypertensive and normotensive rhesus monkeys. PMID- 2551331 TI - Characterization of the binding parameters of 3H-nitrendipine as a Ca2+-channel blocker in the myocardial membrane of the infarction region of mini-pigs. PMID- 2551332 TI - Spontaneous progression of hyperplastic outgrowths of the D1 lineage to mammary tumors: expression of mouse mammary tumor virus and cellular proto-oncogenes. AB - Mammary cancer in mice is characterized by progression through defined stages of preneoplasia, with the most common preneoplastic stage being the hyperplastic alveolar nodule (HAN). We determined the relative levels of RNA expression of various cellular proto-oncogenes and endogenous mouse mammary tumor virus genes in outgrowths and tumors of three sublines of the transplantable D1 HAN preneoplastic outgrowth line. The three sublines differed in relative tumor producing capabilities. Subline D1B produced a high incidence of tumors with short latency periods, whereas sublines D1C and D1D produced low incidences of tumors with long latency periods. No consistent alteration in proto-oncogene expression correlated with relative tumorigenicity, although tumors frequently contained higher levels of one or more proto-oncogene transcripts as compared with preneoplastic tissue. Slightly elevated (2- to 6-fold) levels of different oncogene transcripts were detected in 13 of 17 tumors as compared with outgrowth tissue, including abl (2 tumors), fps (5 tumors), Ha-ras (6 tumors), and Ki-ras (8 tumors). One tumor contained 45 times more Ki-ras-specific RNA than outgrowth tissue because of a comparable amplification of Ki-ras DNA sequences. Elevated levels of Ha-ras occurred more frequently in tumors of a high-incidence subline than in a less-aggressive subline (5/10 vs 1/7), but this difference was not statistically significant. However, consistent changes in MMTV expression accompanied progression from preneoplastic tissues to mammary tumors. All 17 tumors displayed reduced levels of the MMTV-specific long terminal repeat (LTR) transcript (1.6 kb) as compared with HAN tissue; tumors with moderate levels of LTR transcript expressed the 3.8-kb envelope message as well, one not detected in HANs. Expression of the LTR transcript is apparently influenced by factors in addition to the methylation status of endogenous mouse mammary tumor virus genes, which was similar in outgrowths and tumors. As the survey of representative proto oncogenes failed to identify a uniform change between HAN and tumors, it is likely that other genes are involved in tumor progression in the mammary gland. PMID- 2551334 TI - A spin labeling study of pig cardiac tropomyosin. AB - Tropomyosin (TM) extracted from pig cardiac muscle was spin-labeled with 2,2,6,6 tetramethyl-4-(dichlorotriazin)-aminopiperidine-1-oxyl. The ESR spectra of the product (SL-TM) were of a type of weak immobilization. Effects of three means for the denaturation were observed on the above spectra. The ESR spectrum obtained for SL-TM after enzymatic degradation was found to be analogous to that for the label itself in a dilute solution and thereby the quantity of labels bound in SL TM estimated. The Arrhenius plots attained through variable temperature measurement for SL-TM's exhibited two inflexion points (the conformational transition temperatures for TM) around 45 degrees C and 74-75 degrees C, the latter temperature having not been reported in literature so far. However, the enzymatic degradation product from SL-TM behaved quite differently from it in the response to microwave power saturation and temperature variation. PMID- 2551335 TI - Pharmacological study of an irreversible partial agonist of opiate receptors, A alpha-CAO. AB - A-alpha-CAO induces weak analgesia with very short duration in mice and is able to antagonize the analgesic effect of morphine (Mor) up to 3-4 days after a single injection. No tendency of dependence has been observed. It acts as a partial agonist on MVD with Ke value of 9 X 10(-9) mol/L. Its antagonist effect remains after several washes and its agonist effect cannot be reversed by naloxone (Nx), provided the incubation time or the concentration of the agent is sufficient. On isolated GPI, A-alpha-CAO is a pure agonist with IC50 of 5.7 X 10( 10) mol/L; this agonist effect cannot be removed by washing but can be reversed by Nx. On RVD and RbVD, it has antagonist effect against beta-endorphine (beta end) and U50488H, which cannot be washed out easily, and the pA2 are 7.5 and 7.6 respectively. A-alpha-CAO also inhibits the specific binding of 3H-etorphine (3H Etor) to the P2 fraction of the mouse brain membrane with an IC50 of 3.2 X 10(-9) mol/L. The inhibition on the high affinity binding sites of 3H-Etor remains 95% even after 6 washes. PMID- 2551333 TI - Prevalence of nucleoside diphosphate kinase autophosphorylation in human colon carcinoma versus normal colon homogenates. AB - The G-regulatory proteins of adenylate cyclase, tubulin, and the ras oncogene protein product require the production of GTP from ATP in order to exert their effects within the cell. This implies that the activity of nucleoside diphosphate kinase (NDPK) plays a major role in the regulation of cellular events requiring GTP and that the level of activity of this enzyme is critical. This report presents a simple method for trapping a specific isozyme of NDPK in its high energy phosphorylated form (NDPK approximately P) using EDTA and demonstrates that this NDPK approximately P is tenfold higher in malignant colon tumor tissue than in normal colon tissue. This autophosphorylation of the 21,000 and 24,000 Mr subunits of NDPK occurs rapidly at 0 degrees C, will use either [gamma-32P]ATP, [gamma-32P]GTP, or the corresponding 8-azidopurine photoprobes, is intramolecular, displays saturation effects, and is prevented from forming if GTP gamma S is added. Dephosphorylation in the homogenate occurs rapidly upon addition of Mg2+ or any nucleoside-5'-diphosphate. The subunits autophosphorylated in the homogenates are mostly in the soluble phase, and they comigrate with the subunits of pure NDPK from human erythrocytes. Cross-addition of normal and malignant homogenates does not decrease the level of autophosphorylation of NDPK, which indicates that the level of NDPK approximately P may be a quantitative measure of the level of this specific NDPK isozyme form. Assays for NDPK activity show correspondingly elevated levels in the malignant homogenates. Using western blot and photoaffinity labeling techniques, we distinguished the NDPK approximately P subunits from two closely migrating GTP binding proteins. These were identified as the ras gene protein product and a 20,000 Mr protein, which comigrates identically with ADP-ribosylating factor (ARF). The ARF also comigrates in a tight band that is phosphorylated by [gamma 32P]ATP or [gamma-32P]GTP when Mg2+ is present. PMID- 2551337 TI - Cultural differences of the Hispanic patient. PMID- 2551336 TI - Benzodiazepine receptors in isolated adrenal glomerulosa cells. AB - In our experiments the isolated rat adrenal glomerulosa cells displayed peripheral-type benzodiazepine receptors, which could bind to [3H] PK11195 with an apparent equilibrium dissociation constant (KD) of 9.4 +/- 2.8 nmol/L and a maximal binding capacity (Bmax) of 5.6 +/- 1.8 pmol/10(6) cells. The effects of five ligands: PK11195, Ro5-4846, flunitrazepam, diazepam and clonazepam on aldosterone secretion responses of isolated glomerulosa cells to angiotensin II or extracellular potassium ions were observed. The logarithm of EO50 for these ligands as stimulators was well correlated with the logarithm of their Ki value for [3H] PK11195 binding, suggesting that the stimulative effects might be mediated by the benzodiazepine receptor in isolated glomerulosa cells. PMID- 2551338 TI - Possible abuse potential of the NMDA antagonist MK-801. AB - Selective antagonists of the N-methyl-D-aspartate (NMDA) receptor such as MK-801 may have therapeutic potential in the treatment of ischemic brain injury. However, some drugs (e.g. ketamine and phencyclidine) with potent NMDA antagonist properties are also addictive. Intracranial self-stimulation (ICSS) is facilitated by drugs of abuse such as cocaine and amphetamine and thus is useful for screening compounds with potential abuse liability. Low doses of the NMDA antagonist, MK-801 were also found to facilitate ICSS, suggesting that this compound may possess abuse potential. PMID- 2551339 TI - Ontogeny of vasopressin and oxytocin binding sites in the brain of Wistar and Brattleboro rats as demonstrated by lightmicroscopical autoradiography. AB - Vasopressin (VP) and oxytocin (OT) binding sites were localized and quantified in the developing brain of the Wistar, heterozygous (Het) and homozygous (Hom) VP deficient Brattleboro rat using an autoradiographical technique. VP binding sites could be demonstrated from prenatal day 20 onwards in the septum and in the lateral reticular nucleus. Between this and postnatal day 15, VP binding sites appeared in all other brain areas known to contain VP binding sites in adulthood. In the caudate putamen the regional distribution of VP binding changed during development, while in some areas, for instance, the dorsal hippocampus and post cingulate cortex, the concentration of binding sites increased early but decreased with age. Comparison of VP binding between Het and Hom rats showed significant differences in the lateral reticular nucleus during development. Moreover, at postnatal day 15 there was more VP binding in the anterior commissural and suprachiasmatic nucleus and less in the central amygdala, dorsal hippocampus and post cingulate cortex of the Hom rat. This study shows, for the first time, OT binding sites in the developing rat brain. There is a considerable overlap with VP binding in the brain, sometimes with the same developmental pattern, e.g. in the anterior olfactory nucleus and caudate putamen and sometimes with a later appearance, e.g. in the central amygdala and thalamic nuclei. However most areas with VP binding sites did not show OT binding. In some areas only OT binding sites were present, for instance in the islands of Calleja and ventromedial hypothalamus. Similar to some areas with VP binding, OT binding decreased between postnatal day 5 and 15 in the dorsal hippocampus and even completely disappeared in the parietal cortex. The existence of VP binding sites in the Hom rat, together with the only occasional relationship between the previously described ontogeny of VP and OT innervation of the brain and the presently described developmental course of binding sites, indicates that the early expression of binding sites is not initiated by endogenous ligand. However, the setting of the number of VP binding sites has probably been affected by the VP deficiency of the Hom Brattleboro rat. PMID- 2551340 TI - Adenylate cyclase-cyclic AMP-phosphodiesterase system in microdissected brain areas of normotensive and spontaneously hypertensive rats. AB - The topographical distribution of the adenylate-cyclase-cyclic adenosine monophosphate-phosphodiesterase system was investigated in specific brain areas of rats, and compared with spontaneously hypertensive rats. In normotensive animals, brain cyclic adenosine monophosphate levels were quite uniform (between 7.14 and 13.04 pmol/mg protein) with highest concentrations in catecholamine containing cell groups. In contrast, the distribution of basal adenylate cyclase and phosphodiesterase activity is not uniform. The unstimulated adenylate cyclase activity was very low in the striatum and catecholamine-containing cell groups in the medulla oblongata (A1C1-cell groups) and very high in the central gray matter and the cerebellum, where the lowest phosphodiesterase activities were measured. In spontaneously hypertensive rats, altered cyclic adenosine monophosphate levels were found in 17 of 36 brain areas investigated in comparison to those of normotensive rats. Increased concentrations were found in regions which are known to participate in the central regulation of blood pressure (nucleus of the solitary tract, A1C1 catecholaminergic cell groups in the ventrolateral medulla oblongata, locus coeruleus) and in the periaqueductal central gray matter, the hippocampus and the cingulate cortex. Lower levels were measured only in hypothalamic nuclei, especially in the paraventricular and dorsomedial nucleus. No significant differences in basal adenylate cyclase activity were found in spontaneously hypertensive rats compared with Wistar-Kyoto control rats, while phosphodiesterase activity was generally higher in spontaneously hypertensive rats, most significantly in the medulla oblongata. Present data show that characterization of the adenylate cyclase-cyclic adenosine monophosphate phosphodiesterase system helps to localize structural and/or functional differences between the spontaneously hypertensive rat and its normotensive control rat and indicate that more than one functional system is affected in spontaneous hypertension. PMID- 2551341 TI - Diagnostic implication of specific immunoglobulin G patterns of children born to HIV-infected mothers. AB - We analysed HIV-specific immunoglobulin G (IgG) responses to gag and env peptides in infants born to HIV-positive mothers. Questions of interest were whether there are early specific markers for prognosis, and whether any specific IgG is related to the prevention of vertical transmission of infection. Fifty-three children, 0 24 months old and born to HIV-1-infected mothers, were retrospectively divided into two groups based on HIV seroreactivity or non-reactivity at 15 months of age. Their sera were used to find reactivities important in diagnosis and/or prediction of the putative HIV disease. Three important findings emerged. First, a low IgG titer against the very immunodominant penv9 in newborns was found to be associated with rapid progression to AIDS. This difference was clearly reflected in the reactivity to a small peptide representing amino acid (aa) 598-606. The second interesting finding was the putative hypervariable loop on gp120 (especially aa 324-338), reactivity to which was found only in the uninfected group, and was seen in six out of 19 uninfected children under 6 months of age. This specific response was not caused by a generally high total anti-HIV reactivity, and may indicate a role of protective antibodies against vertical transmission. The response to this region in the infected group, on the other hand, was directed to the amino terminal half of the putative loop, in particular peptide 53, aa 304-318. Finally, response to a part of the amino terminal end of P17 was seen in seven out of eight infected children over 6 months of age.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551343 TI - Statistics from the World Health Organization and the Centers for Disease Control. PMID- 2551342 TI - Immunological reactivities of Ghanaian sera with HIV-1, HIV-2, and simian immunodeficiency virus SIVagm. PMID- 2551344 TI - Preoperative imaging in primary hyperparathyroidism. Role of thallium-technetium subtraction scintigraphy. AB - Primary hyperparathyroidism, although often silent clinically, may lead to significant morbidity if it remains untreated. In more than 95% of all cases the cause is a parathyroid adenoma or glandular hyperplasia. Regression of disease follows successful surgical excision of the abnormal parathyroid gland. Recurrent or persistent hyperparathyroidism is most commonly caused by solitary adenomas, which may have an ectopic location. Preoperative localization of lesions may improve postoperative cure rates and decrease morbidity. Thallium T1 201 chloride technetium Tc 99m pertechnetate subtraction scintigraphy was performed on 15 patients with primary hyperparathyroidism. The sensitivity and specificity for detection of abnormal glands were 90% and 95%, respectively. False-positive or false-negative results were minimize by strict adherence to a protocol and by the use of well-defined diagnostic criteria. Because of the superior sensitivity and specificity, this modality should be the primary imaging method of choice for preoperative evaluation of primary hyperparathyroidism. The relative role of other imaging modalities is also discussed. PMID- 2551345 TI - Pathologic quiz case 2. Signet-ring cell adenocarcinoma of the ethmoid sinuses. PMID- 2551346 TI - Investigation of adenylate kinase from Escherichia coli and its interaction with nucleotides by Fourier transform infrared spectroscopy. AB - The secondary structure of adenylate kinase (EC 2.7.4.3) from E. coli was investigated under various conditions using Fourier transform infrared spectroscopy. The overall band contour of the conformation-sensitive amide I mode indicates that in HEPES buffer (pH 7.4) the major structure of the protein is alpha-helical. A more detailed estimate obtained from decomposition of the amide I band into its constituent component bands gives 50% alpha-helix, 26% beta structure, 15% turns and loops, and about 9% nonrepetitive unordered structures. Binding of nucleotide (e.g., ATP) to the donor site decreases the beta-content and shifts the amide I band to higher wavenumbers, whereas binding of nucleotide (e.g., AMP) to the acceptor site does not produce any change in conformation of the protein. These results agree with the protection by ATP and lack of protection by AMP when adenylate kinase is digested with trypsin. The effect of protein denaturing agents and conditions (temperature, high pH, sodium dodecyl sulfate) on changes in the protein conformation as revealed by the conformation sensitive amide I bands is discussed. PMID- 2551347 TI - Evidence for two mechanisms of thrombin-induced platelet activation: one proteolytic, one receptor mediated. AB - The possibility that thrombin-induced platelet reactivity could occur via both a receptor-related and a proteolytic process was examined. Thrombin elicited the formation of considerably more [32P]phosphatidic acid (an index of phospholipase C catalysed phosphoinositide metabolism) than did platelet activating factor, 5 hydroxytryptamine, ADP, and the thromboxane A2 analogue EP171, when these agents were added either alone or in combination. Co-addition of thrombin and EP171 did not evoke significantly more [32P]phosphatide acid than did thrombin alone. The protease inhibitor leupeptin, decreased but did not abolish [32P]phosphatidic acid formation elicited by either thrombin alone or thrombin in combination with EP171. The serine protease, trypsin, stimulated an increase in [32P]phosphatidic acid and this effect was additive with that of EP171. This augmentation by trypsin of EP171-induced [32P]phosphatidic acid formation was inhibited by leupeptin. These results are consistent with the concept that thrombin-induced activation of phospholipase C occurs by two distinct mechanisms: one via proteolysis, which is sensitive to leupeptin, and the other via receptor activation, a process shared by EP171. The individual components of this dual mechanism can be mimicked by the co-addition of a receptor-directed agonist (EP171) and a proteolytic agent (trypsin). PMID- 2551348 TI - In situ hybridization analysis for herpes simplex virus nucleic acids in Alzheimer disease. AB - Histological sections of brain from patients showing evidence of advanced pathology of Alzheimer disease (AD) were examined for the presence of herpes simplex type-1 (HSV-1) nucleic acids by a sensitive in-situ hybridization technique. Samples from neurologically normal patients were examined in parallel. Sensitivity of the assay was verified by the detection of HSV-1 nucleic acids in neurons of trigeminal ganglia taken from cases of AD and normal controls. This indicated that the hybridization reaction was sufficiently sensitive to detect latent HSV-1 infections. Positive hybridization in the brain was only detected in a confirmed case of herpes simplex virus encephalitis. These results appear to confirm previous reports that HSV-1 infection is not directly involved in the pathology associated with AD. PMID- 2551349 TI - Lymphocyte beta-adrenoceptor/effector complex in aging and dementia of the Alzheimer type. AB - We examined density (Bmax), affinity (Kd), and norepinephrine (NE)-stimulated cyclic adenosine monophosphate (cAMP) generation of the beta-adrenergic receptor on lymphocytes in patients with dementia of the Alzheimer type (DAT, n = 13), in normal aged controls (AGED, n = 27), and in young controls (YOUNG, n = 21). Bmax (fmol/mg protein; mean +/- SD) was significantly lower in YOUNG (48.0 +/- 18.7) than in either AGED (64.8 +/- 22.3) or DAT (63.6 +/- 19.5, p less than 0.05). Receptor sensitivity, i.e., the fold increase of stimulated over basal cAMP generation (S/B), was higher in DAT (8.3 +/- 5.2) than in AGED (5.5 +/- 2.3) or YOUNG (5.4 +/- 2.6, p less than 0.07). This difference was due to a significant increase in S/B in female but not male DAT patients compared with AGED (p less than 0.05). There were no differences in levels of circulating NE between DAT and AGED, nor between sexes. Thus, the present data suggest that in this model of the postsynaptic beta-adrenergic receptor/effector complex, receptor sensitivity was increased in women with DAT. PMID- 2551350 TI - Changes of myocardial functions in acute hepatic porphyrias. Role of heme arginate administration. AB - In three patients with attacks of acute intermittent porphyria we found markedly impaired functions of the left ventricle accompanied by ECG changes that returned to normal after administration of heme arginate or cytochrome C administration. In nine patients with other types of porphyria, and in one patient with acute intermittent porphyria not suffering an attack, no significant changes of left ventricle function were observed. The changes seen in patients with an acute attack may be ascribed to acute myocardial hypoxia resulting from defective biosynthesis of heme. We assume that the exogenous supply of heme may improve cellular hypoxia. PMID- 2551351 TI - Familial adenomatous polyposis. AB - Familial adenomatous polyposis is an autosomal dominant disease that includes early development of up to thousands of colorectal adenomas and several extracolonic manifestations. All untreated patients will develop colorectal adenocarcinoma. The treatment of choice is colectomy and ileorectal anastomosis, but restorative proctocolectomy may be considered in selected cases. Polyposis patients treated with ileorectal anastomosis should be followed for life, with regular proctosigmoidoscopy and destruction of new adenomas. Furthermore, regular gastroduodenoscopy should be carried out because of frequent occurrence of premalignant duodenal adenomas. The prognosis is good after prophylactic colectomy in patients without carcinoma. All first degree relatives of affected family members should be examined regularly with proctosigmoidoscopy from the age of ten, and prophylaxis should be organised using a national or regional polyposis register. The recent detection of a specific gene for familial adenomatous polyposis is a long step forward, and several problems may be solved by increasing international cooperation. PMID- 2551352 TI - Sodium pump and calcium channel modulation of Mg-deficiency cardiomyopathy. AB - Hamsters fed a Mg-deficient diet (MD) develop a cardiomyopathy (CM) with foci of myocardial necrosis, calcification, and modest mononuclear and giant cell infiltration. We hypothesize that the lesions are secondary to Ca overload following an increase in myocardial Na due to inhibition of (Na.K)-ATPase and secondary Na-Ca exchange. Nifedipine and digoxin were selected as agents to test this hypothesis. Hamsters were given nifedipine, digoxin, or placebo as sustained release subcutaneous pellets the same day they were started on the MD diet. Animals were killed after 14 days, and MD lesions were quantified in H&E stained slides. Regression analysis showed that nifedipine produced a dose-dependent reduction in lesion abundance (p less than .005) and diameter (p less than .05), while digoxin produced a dose-dependent increase in lesion abundance (p less than .005) and diameter (p less than .005). These results support the hypothesis that MDCM is secondary to Ca overload coupled to inhibition of (Na.K)-ATPase of cardiac myocytes. PMID- 2551353 TI - Demonstration of feline infectious peritonitis virus in conjunctival epithelial cells from cats. A simple and reliable method for clinical veterinary virology screening. AB - A simple technique demonstrating the presence of feline infectious peritonitis virus (FIPV) in cats is described. Smears from the third eyelid (membrana nictitans) were dried on microscope slides, fixed in aetone, and stained using an indirect immunofluorescence method with rabbit-anti-FIPV serum. By this method FIPV-antigen could be demonstrated in the cytoplasma of epithelial cells. The results obtained with this method (M3) were compared with serological and immunohistopathological methods for confirming FIPV-infection in cats. The results obtained by the M3 test were up to 85% in concordance with these other tests. Furthermore, the M3 test was easier and quicker to perform and should be well suited for clinical practice. PMID- 2551354 TI - Interactions between anticancer drugs and other clinically used pharmaceuticals. A review. AB - Drug interactions are increasingly common, since clinical practice is getting more complex with the flood of new drugs. Simultaneously, the increased life expectancy of the population increases the number of individuals likely to receive multiple prescriptions. Cytotoxic drugs generally have a narrow therapeutic index, and are delivered at doses close to toxic levels. Consequently, a slight increase in biological activity caused by an interaction with other concomitantly administered drugs could be deleterious to the patient. Interactions between drugs can sometimes also be used in a positive way, i.e. to increase the therapeutic ratio and overcome drug resistance. Interactions between different cancer treatment modalities have attracted considerable interest. However, much less interest has been devoted to interactions between anticancer drugs and other pharmaceuticals. The purpose of this review is to summarize data about the interactions between anticancer drugs and other clinically used drugs with regard to effects on tumor and toxicity. PMID- 2551355 TI - Treatment of small cell lung cancer. Two-drug versus four-drug chemotherapy and loco-regional irradiation with or without prophylactic cranial irradiation. AB - Fifty-five patients with untreated small cell lung cancer were allocated randomly to receive either a standard 2-drug or a 4-drug chemotherapy regimen. The patients were further randomized to receive or not to receive prophylactic cranial irradiation (PCI) 40 Gy/20 fractions/4 weeks. Each patient also received split-course irradiation against the primary tumour (55 Gy/25 fractions/8 weeks), the mediastinum, and the supraclavicular areas. The standard 2-drug regimen consisted of cyclophosphamide 10 mg/kg i.v. days 1-4 and vincristine 1 mg i.v. days 1 + 4; every 4 weeks. The 4-drug regimen comprised cyclophosphamide 10 mg/kg i.v. days 1-3, vincristine 2 mg i.v. day 1 and 1 mg i.v. day 5, methotrexate 30 mg i.v. days 3 and 5, CCNU 80 mg/m2 i.v. day 2; every 7 weeks. The total treatment time for both protocols was 9 to 12 months. Objective response after 2 cycles of chemotherapy was seen in 46% of patients with the 2-drug regimen and in 56% with the 4-drug regimen. Local radiotherapy increased the response rates to 58% and 90% respectively. The median survival time was 12 months with the 2-drug regimen and 14 months with the 4-drug regimen. The 2-year and 3-year survival rates were 11% and 0% in the 2-drug group and 19% and 15% in the 4-drug group respectively. Toxicity was more severe in the 4-drug group with 4 deaths due to myelosuppression. Altogether, 25 patients received PCI. This did not in any subgroup increase median survival significantly but a reduction of relapses in the central nervous system was seen. Median survival was 13 months with versus 10 months without PCI; 2-year survival rates were 15% and 6% respectively. Morbidity due to PCI did not occur. Although no statistically significant survival advantage could be documented, there was obviously a higher rate of complete responses with multidrug therapy, and longer median duration of remission, median survival and maximal survival. PMID- 2551356 TI - Cyclic AMP does not alter taurine accumulation by rat renal brush border membrane vesicles. AB - Secondary hyperparathyroidism has been attributed to be responsible for the generalized aminoaciduria and phosphaturia of vitamin D deficiency. Since PTH acts in the kidney to generate cAMP, we explored the possibility that its synthetic analog, dbcAMP, would alter the renal transport of taurine (an amino acid lost in the urine in vitamin D deficiency) and Pi. Exposure of renal BBMV prepared from normal and vitamin D-calcium-deficient rats to dbcAMP at concentrations ranging between 10(-4) and 10(-7) M did not alter taurine uptake by these vesicles. Higher dbcAMP concentrations blunted uptake, but these concentrations reduced intravesicular volume, thus representing an artifact of osmolarity. Preincubation of BBMV with dbcAMP for times between 0 and 60 min at 0 or 25 degrees C also did not alter taurine accumulation. Hypotonic lysis of BBMV, allowing entry of the cyclic nucleotide, followed by isotonic resealing did not influence taurine uptake. The addition of potassium fluoride (to inhibit phosphodiesterase activity) and ATP (as an energy source) did not alter taurine accumulation at 60 sec. The uptake of Pi, which is influenced by PTH, was decreased by 25% following exposure to dbcAMP on the internal surface of the vesicle. These data indicate that the taurinuria observed in vitamin D deficiency is unlikely to be related to a PTH-induced increase in intracellular cAMP, unlike the changes in Pi transport, which is sensitive to cyclic nucleotides. PMID- 2551357 TI - [A case of mitochondrial encephalomyopathy with cardiomyopathy due to decreased complex I and IV activities]. AB - We reported a girl with mitochondrial encephalomyopathy, who had various neuromuscular symptoms including dilated cardiomyopathy, generalized convulsions, myoclonus, muscular weakness and growth retardation. Lactate levels in the serum and CSF were elevated. Muscle biopsy showed scattered ragged-red fibers, and complex I (NADH-CoQ reductase) and complex IV (cytochrome c oxidase) were markedly reduced. Although she was treated with coenzyme Q, DL-carnitine and sodium succinate, she died of progressive congestive heart failure at 9 10/12 years of age. PMID- 2551358 TI - Partial protection of oncogene, anti-sense oligodeoxynucleotides against serum nuclease degradation using terminal methylphosphonate groups. AB - Under certain circumstances sequence-specific inhibition of gene expression may be achieved in intact cells using exogenous anti-sense oligodeoxynucleotides. The efficacy of this approach to investigating gene function is limited in part by the rapid serum nuclease mediated degradation of oligodeoxynucleotides in culture media. In order to determine the relative contributions of 3'-exonuclease, 5' exonuclease and endonuclease activity in fetal calf serum to oligodeoxynucleotide destruction, we have tested chimeric N-ras anti-sense sequence molecules protected against exonuclease attack with terminal methylphosphonate diester linkages. An 18-mer with two methylphosphonate diester linkages at the 3' terminus, a 20-mer with two methylphosphonate diester groups at both ends, and the 16-mer 3'-methylphosphonate monoester components of their respective piperidine hydrolysates were totally resistant to venom phosphodiesterase, whereas the 16-mer 3'-hydroxyl components of the hydrolysates were rapidly degraded. Both the chimeric oligodeoxynucleotides and 3'-methylphosphonate monoesters were considerably more stable than normal 3'-hydroxyl oligodeoxynucleotides at 37 degrees C in McCoy's 5A medium containing 15% heat inactivated fetal calf serum. Typically 20-30% of the former (initial concentration 10-100 microM) remained intact at 20 h as compared to the latter which were 88-100% degraded in 4 h and undetectable at 20 h. We conclude that a 3'-phosphodiesterase activity is a predominant nuclease responsible for oligodeoxynucleotide degradation by fetal calf serum, and that for cell culture studies, significant protection of oligodeoxynucleotides may be achieved by incorporating 3'-terminal methylphosphonate diester or even monoester end groups. PMID- 2551359 TI - Maintenance chemotherapy in limited small cell lung cancer: a randomised controlled clinical trial. AB - In a prospective randomised study 68 patients with limited small cell bronchogenic carcinoma were assigned to induction treatment with combined alternating non-cross-resistant chemotherapy plus split course radiotherapy without (NM) or with (M) subsequent maintenance therapy. Induction chemotherapy consisted of cisplatinum and VP16213q. 3 weeks followed by cyclophosphamide, vincristine and methotrexate (CVM)q. 4 weeks. Three courses of this 7-week chemotherapy programme were given. Radiotherapy to the primary lesion of 25 Gy in 13 fractions was given after each of the first and second courses of chemotherapy. Those in complete remission following the induction phase received prophylactic cranial irradiation. Those assigned to maintenance received a further six cycles of CVM after induction. The overall survival of patients randomised to maintenance therapy was significantly inferior to that of those randomised to no maintenance therapy (median survival NM 19.2 vs M 14.1 months, P = 0.05 log rank). Among patients achieving a complete remission of disease on induction therapy those receiving maintenance also showed a trend towards inferior survival (median survival NM 26.8 vs 18.0 months, P = 0.06 log rank). Deaths in each group of patients were predominantly due to tumour progression. The results do not support the use of maintenance chemotherapy after the use of intensive combined therapy induction programmes in the management of limited small cell bronchogenic carcinoma. PMID- 2551360 TI - Human primary brain tumour metabolism in vivo: a phosphorus magnetic resonance spectroscopy study. AB - Magnetic resonance spectroscopy was used to study intracellular pH and compounds which contain phosphorus in normal human brain and primary brain tumours non invasively. In normal subjects (n = 7) intracellular pH (pHi) of the brain was 7.03 +/- 0.02 (mean +/- s.e.m.). The pHi did not vary between superficial (2 cm, majority grey matter) and deep brain (5 cm, majority white matter). The relative concentrations of phosphocreatine (PCr) and phosphomonoesters (PME) to ATP were also constant with depth. The relative concentration of phosphodiesters (PDE) increased from superficial to deep in normal brain. The astrocytomas (n = 7, grade II-IV) were significantly more alkaline (pHi = 7.08 +/- 0.03), and contained more PCr and PME, with respect to ATP, than normal brain at similar depth. The meningiomas (n = 4) were also more alkaline (pHi = 7.19 +/- 0.02) with a raised PME level but reduced PCr. The reduction in meningioma PCr may be due to the significant necrosis (greater than 20%) seen in the surgical biopsies. No significant necrosis was seen in the gliomas. Previous in vitro studies suggest that increased PME may be due to accumulation of phosphoethanolamine (PE), a phospholipid precursor. These results suggest that human primary brain tumours characteristically are more alkaline with increased PME than normal brain. PMID- 2551361 TI - Preparation and characterization of a [14C]cellulose suitable for human metabolic studies. AB - In six normal subjects administered 5 microCi of an oral dose of a commercially available 14C-labelled cellulose, significant amounts of 14CO2 were detected in expired air within 30 min, suggesting that other 14C-labelled non-cellulosic material was present. Chemical and microscopical examination confirmed that starch was the principal contaminant. The commercial preparation was purified using amyloglucosidase (EC 3.2.1.3) digestion following gelatinization of the starch by autoclaving. Subsequent administration of the purified cellulose to a further six normal subjects decreased the expired air 14CO2 during the subsequent 10 h from 13.0 (SD 4.0) to 4.1 (SD 1.9)%. Administration of the purified product to a further group of four normal subjects, before and after a regimen of increased dietary fibre, showed a cumulative increase in expired 14CO2 over 24 h from 7.9 (SD 1.1) to 12.1 (SD 2.6)% on fibre. In six ileostomy subjects the cumulative excretion of 14CO2 was greatly decreased compared with normal controls (3.0 (SD 1.14) and 10.5 (SD 3.9)% respectively). In constipated subjects expired 14CO2 continued beyond 48 h, in contrast to normal subjects where expired 14CO2 at this time was negligible. PMID- 2551362 TI - Intestinal zinc transfer by everted gut sacs from rats given diets containing different amounts and types of dietary fibre. AB - Two experiments were carried out in which rats were offered diets containing different amounts and types of dietary fibre, i.e. commercial stock diet and three semi-purified diets containing no fibre, 200 g wheat bran or 200 g pectin/kg. Dietary inclusion of fibre, and especially pectin, stimulated large bowel fermentation, as indicated by caecal hypertrophy and reduced caecal pH. After 3 weeks, mucosal:serosal zinc transfer and Zn accumulation by tissue were measured using the everted-gut-sac technique. In Expt 2, incubations were carried out in the presence and absence of 0.25 mM-ouabain to assess the importance of transfer by Na+,K+-ATPase-dependent mechanisms, and some observations on glucose transport were also made. Ouabain reduced rates of transfer of both Zn and glucose and also tissue Zn accumulation. There were no significant differences in rates of Zn transfer by everted sacs from duodenal, ileal and colonic sites, but accumulation of Zn by tissue was a more important fate than transfer across the serosal surface, and accumulation by duodenal tissue was approximately twice as great as by other tissues. Mucosal:serosal transfer of glucose by ileal tissue was much more sensitive to ouabain than was Zn transfer. Previous diet appeared to alter the capacity of the intestinal tissue to transfer Zn, with the highest rates of transfer being by colonic tissue from pectin-fed rats. PMID- 2551363 TI - The availability of zinc in endosperm, whole grain and bran-enriched wheat crispbreads fed to rats on a Zn-deficient diet. AB - The hypothesis that factors such as dietary fibre and phytate in wheat bran limit the availability of Zn was tested in growing rats fed on low-Zn diets with different wheat crispbreads as the major source of Zn. Six groups of six weanling male rats each were fed on 5 parts semi-synthetic Zn-deficient diet and 1 part wheat-endosperm crispbread for 1 week. At the beginning of the second week, the crispbread in the diet of five groups was exchanged for crispbread made using one of the following wheat flours: (1) whole grain, (2) bran-enriched whole grain, (3) endosperm with Zn added to the whole-grain level, (4) endosperm with Zn added to the bran-enriched level, (5) whole grain with Zn added to the bran-enriched level. These diets were given ad lib. together with deionized water for 2.5 weeks. The relative absorption of Zn was lowest from the three non-supplemented diets (75-82%). All the added Zn was absorbed. As appetite, body-weight increase, Zn absorption, Zn retention and the Zn concentrations in serum and bone differed only slightly among groups fed on diets with similar Zn concentrations, it is concluded that factors such as dietary fibre or phytate in wheat bran limit the availability of Zn in wheat crispbreads very little when all the Zn is needed for growth and development in rats. PMID- 2551364 TI - Mutagenesis of the cysteines in the metalloregulatory protein MerR indicates that a metal-bridged dimer activates transcription. AB - Bacterial resistance to mercury(II) compounds is controlled by the metalloregulatory MerR protein, a transcriptional repressor and a mercuric ion dependent activator of the mer operon. Site-directed mutagenesis of all four cysteine residues in the Tn501 MerR protein has led to the specific replacement of C82, C115, and C117 with alanine and of C126 with serine. Mutation of C82 and C126 abolishes transcriptional activation in vivo while mutation of C115 and C117 leads to a slight increase and dramatic decrease in transcriptional activation, respectively. All four mutants are competent, to varying degrees, to repress mer transcription. Characterization of the four purified mutant proteins in vitro demonstrates that only the C126S MerR mutant is most notably deficient in stoichiometric Hg(II) binding. All four mutant proteins possess similar DNA binding properties, and the C82 mutant is most affected in the ability to form stable dimers. Given an observed stoichiometry of one Hg(II) per MerR dimer, it is likely that the transcriptionally activating MerR species is a metal-bridged dimer. It is most likely that one C126 per subunit provides high-avidity bidentate ligation to Hg(II), but it remains possible that C82 may be a secondary Hg(II) ligand (e.g., in a tetracoordinate thiol ligation array). PMID- 2551365 TI - G protein-effector coupling: interactions of recombinant inhibitory gamma subunit with transducin and phosphodiesterase. AB - A bacterial expression vector for the inhibitory gamma subunit of retinal rod phosphodiesterase has been constructed by inserting a mouse gamma cDNA into pUC19. Escherichia coli 222 transformed with this plasmid produces a 12-kDa recombinant protein consisting of 18 additional amino acids attached to the amino terminus of gamma. The fusion protein, designated beta-gal-gamma, has been refolded into an active form in formic acid and partially purified by gel filtration chromatography. Despite a large extended sequence at the amino terminus, beta-gal-gamma is able to inhibit the activity of trypsin-activated phosphodiesterase, bind tightly to the catalytic alpha beta subunits, and interact with the alpha subunit of transducin in a nucleotide-dependent manner. The availability of large quantities of active bacterial gamma, together with the ability to change its primary structure by site-directed mutagenesis, promises to provide considerable new information on the interaction between transducin and phosphodiesterase, as well as insights into the molecular mechanism of G protein effector coupling. PMID- 2551366 TI - Effect of antineoplastic agents on the DNA cleavage/religation reaction of eukaryotic topoisomerase II: inhibition of DNA religation by etoposide. AB - Beyond its essential physiological functions, topoisomerase II is the primary cellular target for a number of clinically relevant antineoplastic drugs. Although the chemotherapeutic efficacies of these drugs correlate with their abilities to stabilize the covalent topoisomerase II-DNA cleavage complex, their molecular mechanism of action has yet to be described. In order to characterize the drug-induced stabilization of this enzyme-DNA complex, the effect of etoposide on the DNA cleavage/religation reaction of Drosophila melanogaster topoisomerase II was studied. Under the conditions employed, etoposide increased levels of enzyme-mediated double-stranded DNA cleavage 5-6-fold and single stranded cleavage approximately 4-fold. Maximal stimulation was observed at 80 100 microM etoposide with 50% of the maximal effect at approximately 15 microM drug. By employing a topoisomerase II mediated DNA religation assay [Osheroff, N. & Zechiedrich, E.L. (1987) Biochemistry 26, 4303-4309], etoposide was found to stabilize the enzyme-DNA cleavage complex (at least in part) by inhibiting the enzyme's ability to religate cleaved DNA. Moreover, in order for the drug to affect religation, it has to be present at the time of DNA cleavage. PMID- 2551367 TI - Double-stranded DNA cleavage/religation reaction of eukaryotic topoisomerase II: evidence for a nicked DNA intermediate. AB - The DNA cleavage reaction of eukaryotic topoisomerase II produces nicked DNA along with linear nucleic acid products. Therefore, relationships between the enzyme's DNA nicking and double-stranded cleavage reactions were determined. This was accomplished by altering the pH at which assays were performed. At pH 5.0 Drosophila melanogaster topoisomerase II generated predominantly (greater than 90%) single-stranded breaks in duplex DNA. With increasing pH, less single stranded and more double-stranded cleavage was observed, regardless of the buffer or the divalent cation employed. As has been shown for double-stranded DNA cleavage, topoisomerase II was covalently bound to nicked DNA products, and enzyme-mediated single-stranded cleavage was salt reversible. Moreover, sites of single-stranded DNA breaks were identical with those mapped for double-stranded breaks. To further characterize the enzyme's cleavage mechanism, electron microscopy studies were performed. These experiments revealed that separate polypeptide chains were complexed with both ends of linear DNA molecules generated during cleavage reactions. Finally, by use of a novel religation assay [Osheroff, N., & Zechiedrich, E. L. (1987) Biochemistry 26, 4303-4309], it was shown that nicked DNA is an obligatory kinetic intermediate in the topoisomerase II mediated reunion of double-stranded breaks. Under the conditions employed, the apparent first-order rate constant for the religation of the first break was approximately 6-fold faster than that for the religation of the second break. The above results indicate that topoisomerase II carries out double-stranded DNA cleavage/religation by making two sequential single-stranded breaks in the nucleic acid backbone, each of which is mediated by a separate subunit of the homodimeric enzyme. PMID- 2551368 TI - Strand specificity of the topoisomerase II mediated double-stranded DNA cleavage reaction. AB - The strand specificity of topoisomerase II mediated DNA cleavage was analyzed at the nucleotide level by characterizing the enzyme's interaction with a strong DNA recognition site. This site was isolated from the promoter region of the extrachromosomal rRNA genes of Tetrahymena thermophila and was recognized by type II topoisomerases from a variety of phylogenetically diverse eukaryotic organisms, including Drosophila, Tetrahymena, and calf thymus. When incubated with this site, topoisomerase II was found to introduce single-stranded breaks (i.e., nicks) in addition to double-stranded breaks in the nucleic acid backbone. Although the nucleotide position of cleavage on both the noncoding and coding strands of the rDNA remained unchanged, the relative ratios of single- and double stranded DNA breaks could be varied by altering reaction conditions. Under all conditions which promoted topoisomerase II mediated DNA nicking, the enzyme displayed a 3-10-fold specificity for cleavage at the noncoding strand of its recognition site. To determine whether this specificity of topoisomerase II was due to a faster forward rate of cleavage of the noncoding strand or a slower rate of its religation, a DNA religation assay was performed. Results indicated that both the noncoding and coding strands were religated by the enzyme at approximately the same rate. Therefore, the DNA strand preference of topoisomerase II appears to be embodied in the enzyme's forward cleavage reaction. PMID- 2551369 TI - Quinoprotein D-glucose dehydrogenase of the Acinetobacter calcoaceticus respiratory chain: membrane-bound and soluble forms are different molecular species. AB - Acinetobacter calcoaceticus is known to contain soluble and membrane-bound quinoprotein D-glucose dehydrogenases, while other oxidative bacteria contain the membrane-bound enzyme exclusively. The two forms of glucose dehydrogenase were believed to be the same enzyme or interconvertible forms. Previously, Matsushita et al. [(1988) FEMS Microbiol. Lett 55, 53-58] showed that the two enzymes are different with respect to enzymatic and immunological properties, as well as molecular weight. In the present study, we purified both enzymes and compared their kinetics, reactivity with ubiquinone homologues, and immunological properties in detail. The purified membrane-bound enzyme had a molecular weight of 83,000, while the soluble form was 55,000. The purified enzymes exhibited totally different enzymatic properties, particularly with respect to reactivity toward ubiquinone homologues. The soluble enzyme reacted with short-chain homologues only, whereas the membrane-bound enzyme reacted with long-chain homologues including ubiquinone 9, the native ubiquinone of the A. calcoaceticus. Furthermore, the two enzymes were distinguished immunochemically; the membrane bound enzyme did not cross-react with antibody raised against the soluble enzyme, nor did the soluble enzyme cross-react with antibody against the membrane-bound enzyme. Thus, each glucose dehydrogenase is a molecularly distinct entity, and the membrane-bound enzyme only is coupled to the respiratory chain via ubiquinone. PMID- 2551370 TI - Redox protein electron-transfer mechanisms: electrostatic interactions as a determinant of reaction site in c-type cytochromes. AB - The effect of ionic strength on the rate constant for electron transfer has been used to determine the magnitude and charge sign of the net electrostatic potential which exists in close proximity to the sites of electron transfer on various c-type cytochromes. The negatively charged ferricyanide ion preferentially reacts at the positively charged exposed heme edge region on the front side of horse cytochrome c and Paracoccus cytochrome c2. In contrast, at low ionic strength, the positively charged cobalt phenanthroline ion interacts with the negatively charged back side of cytochrome c2, and at high ionic strength at a positively charged site on the front side of the cytochrome. With horse cytochrome c, over the ionic strength range studied, cobalt phenanthroline reacts only at a positively charged site which is probably not at the heme edge. These inorganic oxidants do not react at the relatively uncharged exposed heme edge sites on Azotobacter cytochrome c5 and Pseudomonas cytochrome c-551, but rather at a negatively charged site which is away from the heme edge. The results demonstrate that at least two electron-transferring sites on a single cytochrome can be functional, depending on the redox reactant used and the ionic strength. Electrostatic interactions between charge distributions on the cytochrome surface and the other reactant, or interactions involving uncharged regions on the protein(s), are critical in determining the preferred sites of electron transfer and reaction rate constants. When unfavorable electrostatic effects occur at a site near the redox center, less optimal sites at a greater distance can become kinetically important. PMID- 2551371 TI - Application of fluorescence photobleaching recovery to assess complex formation between the two envelope proteins of Sendai virus in membranes of fused human erythrocytes. AB - Fusion of human erythrocytes by Sendai virions is accompanied by lateral mobilization of the viral envelope proteins (F, the fusion protein, and HN, the hemagglutinin/neuraminidase protein) in the target cell membrane; the dynamic parameters characterizing the lateral diffusion of F and HN in the fused cell membrane are identical [Henis, Y. I., & Gutman, O. (1987) Biochemistry 26, 812 819; Aroeti, B., & Henis, Y. I. (1988) Biochemistry 27, 5654-5661]. This identity raised the possibility that F and HN diffuse together in the cell membrane in mutual heterocomplexes. In order to investigate the possible formation of F-HN complexes in the target cell membrane, which could be important for the fusion process mediated by the viral envelope proteins, we combined fluorescence photobleaching recovery (FPR) measurements of the lateral mobility of the viral glycoproteins with antibody-mediated cross-linking of F or HN. After fusion, one viral glycoprotein type was immobilized by cross-linking with highly specific bivalent polyclonal IgG. The other glycoprotein type was labeled with fluorescence monovalent Fab' fragments that do not induce cross-linking, and its mobility was measured by FPR. Neither the mobile fraction nor the lateral diffusion coefficient of the Fab'-labeled viral glycoproteins was affected by immobilization of the second viral envelope protein, demonstrating that F and HN diffuse independently in the target cell membrane and are not associated in mutual complexes. PMID- 2551372 TI - Functional reconstitution of the glycine receptor. AB - The functional reconstitution of the chloride channel coupled glycine receptor is described. Glycine receptors were purified from the cholate extract of rat spinal cord membranes by affinity chromatography and incorporated into phospholipid vesicles by the addition of phosphatidylcholine and removal of detergent by gel filtration. The reconstituted vesicles showed the same polypeptide composition as the purified receptor (proteins of Mr 48,000 and 58,000). The pharmacological characteristics of the glycine receptor were also preserved in the proteoliposomes, as demonstrated by the displacement of [3H]strychnine binding by several glycinergic ligands and by photoaffinity labeling experiments. In order to observe functional responses (i.e., specific agonist-induced anion translocation), we have developed an assay based on the fluorescence quenching of an anion-sensitive entrapped probe, SPQ [6-methoxy-N-(3-sulfopropyl)quinolinium]. Reconstituted vesicles were loaded with the fluorescent probe during a freeze thaw-sonication cycle in the presence of added liposomes containing cholesterol. In such a reconstituted system, glycine receptor agonists are able to increase the rate of anion influx into the vesicles. The action of agonists is blocked by the simultaneous presence of strychnine or other glycine antagonists. Our results show that the purified 48,000- and 58,000-dalton polypeptides reconstituted into phospholipid vesicles can bind ligands and promote specific ion translocation in a way similar to the glycine receptor in its native environment. PMID- 2551373 TI - Resonance Raman studies of CuA-modified cytochrome c oxidase. AB - Modification of the CuA site in mammalian cytochrome c oxidase has been used to elucidate the functional role of this center in the catalytic cycle of the enzyme. Both heat treatment in detergents and chemical modification by p (hydroxymercuri)benzoate (pHMB) convert CuA to a lower potential type II center and effectively remove the site from the electron-transfer pathway during turnover. In this study, resonance Raman spectroscopy has been employed to investigate the effects of these CuA modifications on the heme active sites. The Raman data indicate some environmental perturbation of the heme a3 chromophore in the modified derivatives. Only pHMB modification and SB-12 heat treatment produced significant effects in the Raman spectra of the fully reduced enzyme. These perturbations are much less evident in the spectra obtained within 10 ns of CO photolysis from the fully reduced species of the modified enzymes. Transient Raman studies further indicate that the half-time for CO religation in the modified enzymes is quite similar to that of the native protein. PMID- 2551374 TI - Characterization of multiple forms of the Ah receptor: comparison of species and tissues. AB - Biochemical and toxic responses to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) appear to be mediated via the Ah receptor, a gene-regulatory protein that, like steroid hormone receptors, undergoes a ligand-dependent acquisition of affinity for nuclei and DNA. Since responses to TCDD are highly species- and tissue specific, we compared DNA-binding properties of Ah receptor from several tissues of rat, C57BL/6 mouse, hamster, and guinea pig, using DNA-Sepharose chromatography. Hepatic cytosol from all species contained TCDD.receptor complexes that eluted at approximately 0.15 (peak 1) and approximately 0.33 M NaCl (peak 2). The relative proportions of these forms as well as of TCDD receptor that did not bind to DNA (i.e., was present in flowthrough fractions) varied among species. In each case, the yield of the higher affinity form (peak 2) increased with time or temperature of incubation. Cytosol from lung, thymus, kidney, and testis contained the same two forms; peak 2 was the major DNA-binding form only in thymus. In KCl extracts of hepatic nuclei from animals treated with [3H]TCDD, only the higher affinity form (peak 2) was found. Peak 1 isolated from cytosol by DNA-Sepharose and incubated with hepatic cytosol from D2 mouse (which contains no detectable receptor) transformed into peak 2, suggesting that these two forms are different conformations of the same protein. Sucrose density gradient and gel filtration analyses of peaks 1 and 2 isolated from DNA-Sepharose indicated that (i) the untransformed form (peak 1) was smaller than the unoccupied and the transformed forms, (ii) 0.4 M KCl in the density gradients had little effect on these isolated forms, and (iii) nuclear receptor sedimented like peak 2. On the basis of these results, we hypothesize that the Ah receptor exists in several forms: When occupied, it has no affinity for DNA. Ligand binding initially yields a smaller form with low DNA affinity (i.e., peak 1), as well as, in some cases, a form with no DNA affinity (flowthrough fractions); further incubation in the presence of cytosolic factor(s) induces a change conferring higher DNA affinity and faster sedimentation (i.e., peak 2). The latter form is likely the transcriptionally active form in vivo. Species and tissue differences in this scheme are quantitative rather than qualitative. PMID- 2551375 TI - A developmentally regulated gene of trypanosomes encodes a homologue of rat protein-disulfide isomerase and phosphoinositol-phospholipase C. AB - We have isolated and characterized a developmentally regulated gene in Trypanosoma brucei, arbitrarily termed BS2. BS2 mRNA is substantially more abundant in bloodstream-form trypanosomes than in procyclic culture forms. Its nucleotide sequence reveals a single contiguous open-reading frame of 497 codons and is predicted to encode a protein of approximately 55.5 kilodaltons. A search of the NBRF protein data base revealed that within the predicted amino acid sequence are two of the evolutionarily conserved redox sites typified by thioredoxin of bacteria. Of this family of proteins, the recently sequenced rat genes encoding protein-disulfide isomerase (PDI) and form I phosphoinositide specific phospholipase C (PIPLC) showed homology extending over the length of all three proteins (i.e., between BS2, PDI, and PIPLC). Although this homology includes the acidic C-terminus characteristic of proteins localized to the lumen of the endoplasmic reticulum, the BS2 product is predicted to possess multiple sites for N-linked glycosylation while PDI and PIPLC have none. Possible roles of the BS2 gene product in trypanosome physiology are discussed. PMID- 2551376 TI - Effect of ethylene glycol on the interaction of different myosin subfragment 1.nucleotide complexes with actin. AB - To elucidate the structure of the cross-bridge intermediates in the actomyosin ATPase cycle, several laboratories have added both ethylene glycol and AMP-PNP to muscle fibers. These studies suggested that ethylene glycol shifts the structure of myosin.AMP-PNP toward the weak-binding conformation, i.e., toward the structure of myosin.ATP. Since only the weak-binding conformation of myosin subfragment 1 (S-1) binds with no apparent cooperativity to the troponin tropomyosin-actin complex (regulated actin), we used this as a probe to examine the conformation of various S-1.nucleotide complexes in ethylene glycol. Our results show that ethylene glycol markedly weakens the binding strength of S-1, S 1.ADP, and S-1.AMP-PNP to actin but has almost no effect on the binding strength of S-1.ATP. As in muscle fibers, at 40% ethylene glycol, the binding strength of S-1.AMP-PNP to actin becomes very similar to the binding strength of S-1.ATP. In the presence of troponin-tropomyosin, the binding of S-1.AMP-PNP to actin shows no apparent cooperativity in 40% ethylene glycol. Therefore, our results confirm that ethylene glycol shifts the structure of the myosin.AMP-PNP toward the weak binding conformation. However, our results also suggest that ethylene glycol has a direct effect on the regulated actin complex. This is shown by the fact that ethylene glycol markedly increases the cooperative binding of S-1.ADP to regulated actin both in the presence and in the absence of Ca2+.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551377 TI - Binding of mithramycin to DNA in the presence of second drugs. AB - Comparative DNA equilibrium binding studies with mithramycin (MTR) and ethidium bromide in the presence and in the absence of second drugs were investigated by spectral titrations. Unusual curvatures (in contrast to those due to neighbor exclusion or anticooperativity) are found in the Scatchard plots of MTR-DNA titrations in the presence of netropsin, a minor-groove binder. Parallel studies with ethidium bromide indicate that although the presence of netropsin significantly reduces the binding ability of ethidium, no unusually curved Scatchard plots are obtained. The unusual curvature exhibited by the Scatchard plots of MTR titrations in the presence of netropsin indicates that the binding of netropsin greatly affects the MTR binding to DNA and can be simulated by an explicit incorporation of the second drug-DNA interaction in the binding formalism. Since netropsin is a minor-groove binder, its interference with the binding of MTR is in accord with the notion that MTR also binds at this groove. The observation of negligible effects on the DNA binding ability of MTR in the presence of either a major-groove or a phosphate group binder lends further support to this conclusion. Consistent with its guanine specificity, studies with synthetic polynucleotides suggest that MTR exhibits negligible affinity for poly(dA-dT).poly(dA-dT) or poly(dA).poly(dT).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551378 TI - Cytochrome c at charged interfaces. 1. Conformational and redox equilibria at the electrode/electrolyte interface probed by surface-enhanced resonance Raman spectroscopy. AB - The structure and the electron-transfer properties of cytochrome c (cyt c) absorbed on a silver electrode were analyzed by surface-enhanced resonance Raman spectroscopy. It was found that the absorbed cyt c exists in various conformational states depending on the electrode potential. In state I the native structure of the heme protein is fully preserved and the redox potential (+0.02 V vs saturated calomel electrode) is close to the value for cyt c in solution. In state II the heme iron exists in a mixture of five-coordinated high-spin and six coordinated low-spin configurations. It had been shown that these configurations form a thermal equilibrium [Hildebrandt, P., & Stockburger, M. (1986) J. Phys. Chem. 90,6017]. It is demonstrated that these equilibria strongly depend on the charge distribution within the electrical double layer of the silver electrode/electrolyte interface, indicating that the changes in the coordination shell are induced by electrostatic interactions. The structural alterations in state II are apparently restricted to the heme crevice, which assumes an open conformation compared to the close structure in state I. This leads to a strong decrease of the redox potentials, which were determined to be -0.31 and -0.41 V for the five-coordinated high-spin and six-coordinated low-spin configurations, respectively. On the other hand, gross distortions of the protein structure can be excluded since the reversible proton-induced conformational change of cyt c as found in solution at low pH also takes place in state II of the absorbed cyt c. The linkage of cyt c molecules to the surface is mediated by charged amino acid groups, and it depends on the potential which groups are thermodynamically favored to form such a molecular binding site. The conformational states I and II, which are in potential-dependent equilibrium, therefore refer to two different molecular binding sites. At potentials below zero charge (less than approximately -0.6 V) a rapid denaturation of the absorbed cyt c is noted, which is reflected by drastic and irreversible changes in the surface-enhanced resonance Raman spectrum. Our results are discussed on the background of previous electrochemical studies of cyt c at electrodes. PMID- 2551379 TI - Cytochrome c at charged interfaces. 2. Complexes with negatively charged macromolecular systems studied by resonance Raman spectroscopy. AB - We have analyzed the structure of cytochrome c (cyt c) bound in a variety of complexes in which negatively charged molecular groups interact with the positively charged binding domain around the heme crevice of cyt c. Using resonance Raman spectroscopy, we could demonstrate that these interactions induce the same conformational changes as they were observed in the surface-enhanced resonance Raman experiments of cyt c adsorbed on the Ag electrode [Hildebrandt & Stockburger (1989) Biochemistry (preceding paper in this issue)]. When cyt c is bound to (As4W40O140)27-, state II is stabilized, whereas in complexes with phosvitin and cytochrome b5 state I is formed. The complexes with phospholipid vesicles and inverted micelles reveal a mixture of both states. It is suggested that these systems as well as cyt c adsorbed on the Ag electrode may be regarded as model systems for the physiological complexes of cyt c with cytochrome oxidase and cytochrome reductase. On the basis of our findings it is proposed that the biological electron-transfer reactions are controlled by electric field induced conformational transitions of cyt c upon complex formation with its physiological redox partners. PMID- 2551380 TI - Competitive binding of ATP and the fluorescent substrate analogue 2',3'-O-(2,4,6 trinitrophenylcyclohexadienylidine) adenosine 5'-triphosphate to the gastric H+,K+-ATPase: evidence for two classes of nucleotide sites. AB - ATP and the fluorescent substrate analogue TNP-ATP bind competitively to the gastric H,-K-ATPase. Substrate and product completely reverse the fluorescence enhancement caused by TNP-ATP binding to the enzyme. The fluorophore is displaced monophasically from apoenzyme. However, ATP displaces TNP-ATP from the Mg2+ quenched state in two steps of equal amplitude. The midpoints of the titrations differ by more than 2 orders of magnitude. The estimated substrate constants are in reasonable agreement with published Michaelis constants. TNP-ATP is not a substrate for the H,K-ATPase. The fluorophore prevents phosphorylation by ATP and competitively inhibits the K+-stimulated pNPPase and ATPase activities of the enzyme. Ki is approximately the same for both hydrolytic activities and consistent with the Kd of TNP-ATP measured directly. Km for pNPP is 1.48 +/- 0.15 mM. Two Michaelis constants are required to fit the ATPase data: Km1 = 0.10 +/- 0.01 mM and Km2 = 0.26 +/- 0.05 mM. PMID- 2551381 TI - Variation of DNA sequence specificity of DNA-oligopeptide binding ligands related to netropsin: imidazole-containing lexitropsins. AB - CD binding studies of nonintercalative oligopeptides related to netropsin, named lexitropsins, have been carried out with synthetic duplex DNAs and natural DNA. While netropsin possesses a high dA.dT sequence specificity, these ligands show a progressive lowering of the ability to bind to dA.dT basepairs in DNA and a dramatic reduction of the sequence specificity seen at high salt concentration due to a replacement of pyrrole moieties by imidazoles. This variation in DNA sequence specificity of lexitropsins is mirrored in corresponding large differences in the template inactivation of poly(dA-dT).poly(dA-dT) in the RNA polymerase reaction by these drugs. The presence of imidazole permits binding of the oligopeptide to dG.dC pairs, which is most effective for the triimidazole peptide. Results at increasing salt concentration reveal, however, that a tight binding to pure dG.dC sequences does not occur. A proper sequence containing dG.dC and dA.dT pairs is supposed to be required for a higher specificity. The CD data accord well with previously reported melting studies and are in favor of recent theoretical results suggesting that the diminished AT preference may be due to an increase in the complexation energy with the dG.dC pairs. PMID- 2551382 TI - Study by NMR of the mode of action of monensin on Streptococcus faecalis de energized and energized cells. AB - Streptococcus faecalis was used as a bacterial model for studying the mode of action of monensin by NMR investigations. Experiments were carried out in two states, characterized by several complementary methods: (i) the resting (de energized) cell which was considered as an inert biological membrane, on which cationic transport induced by the ionophore alone can be investigated; (ii) the active (energized) cell where the ionophore-sensitive response of the living organism, particularly the cation pumps and the glycolysis, is probed. Studies of resting cells were performed, with changing external ionic concentrations, in the presence of monensin, which is preferentially a sodium carrier. Internal and external Na+ and H+ were followed by corresponding 23Na and 31P (inorganic phosphate) NMR resonances, K+ fluxes were measured by atomic absorption. It was shown that the induced cationic movements were linked to the existing ionic gradients for K+ and Na+. 31P and 13C NMR spectra for the intermediary metabolites detected in active cells showed that glycolysis is dramatically modified in the presence of monensin. PMID- 2551383 TI - Regulation of oxytocin-induced phosphoinositide breakdown in adipocytes by adenosine, isoproterenol and insulin. AB - In rat adipocytes, the breakdown of phosphoinositides labelled by a 3 h incubation with [3H]inositol resulted in the accumulation of labelled inositol mono-, bis- and trisphosphates in the presence of oxytocin, vasotocin or vasopressin. Oxytocin at a concentration of 1 nM markedly increased phosphoinositide breakdown. Incubation of adipocytes both during the 3 h labelling and the 10 min breakdown period in a low adenosine medium (presence of adenosine deaminase) or high adenosine medium (presence of 0.1 microM N6 (phenylisopropyl)adenosine) (PIA) did not affect basal or ligand-stimulated phosphoinositide breakdown. The addition of 1 microM PIA only during the measurement of phosphoinositide breakdown variably stimulated basal breakdown but significantly potentiated that due to oxytocin. Isoproterenol similarly had little effect on basal but inhibited oxytocin stimulation of phosphoinositide breakdown. Insulin did not affect basal or ligand-stimulated phosphoinositide breakdown in the low or high adenosine medium. However, in adipocytes incubated in the absence of added adenosine deaminase or PIA, insulin stimulated basal accumulation of inositol phosphates by about 20% and inhibited that due to oxytocin by about 20%. There was no significant effect of insulin on the stimulation by vasopressin or vasotocin of phosphoinositide breakdown. These results indicate that, in adipocytes, phosphoinositide breakdown stimulated by oxytocin is enhanced by adenosine, inhibited by isoproterenol and, under some conditions is inhibited by insulin. PMID- 2551384 TI - The effects of thyrotropin-releasing hormone and potassium depolarization on phosphoinositide metabolism and cytoplasmic calcium in bovine pituitary cells. AB - Addition of thyrotropin-releasing hormone (TRH) (10 nM to 10 microM) to bovine anterior pituitary cells labelled with [3H]inositol decreased the radioactivity in inositol-containing lipids and increased it in inositol phosphates. TRH also increased the cytoplasmic calcium concentration biphasically. At TRH concentrations below 10 nM, the increase was sustained and sensitive to inhibitors of calcium influx through voltage-gated channels, whereas concentrations over 10 nM elicited in addition a rapid transient increase in calcium, which was relatively insensitive to such inhibition. Incubation of the cells in medium containing 25 mM KCl increased the cytoplasmic calcium concentration by stimulating influx through voltage-gated channels, and markedly enhanced the initial transient increase of calcium seen at TRH concentrations above 10 nM. It did not affect the generation of InsP3 and it also enhanced the calcium response to ionomycin. It is suggested that stimulation of calcium entry through voltage-gated channels can increase the amount of calcium available for mobilisation by TRH. PMID- 2551386 TI - Triphasic reduction of bH and the absence of equilibration at the i-site of bc1 complex. AB - The concept of a stabilized semiquinone radical forming the basis of a two electron gate has long been familiar in the context of the quinone-reducing site in photosynthetic systems and has already been suggested to play a role at the i site of the bc1-type complexes. It is here pointed out that this concept is sufficient to explain the so-called triphasic reduction kinetics of cytochrome bH, in which cytochrome bH goes partially reduced, is reoxidized and then goes fully reduced. The rate constants for binding and unbinding of quinone, quinol and semiquinone at the i-site are discussed, and a kinetic model featuring slow release of the i-site semiquinone is shown to display many features of the kinetics of electron transfer at the i-site. PMID- 2551385 TI - Thiobacillus ferrooxidans cytochrome c-552: purification and some of its molecular features. AB - Soluble cytochrome c-552 was purified from Thiobacillus ferrooxidans to an electrophoretically homogeneous state. The cytochrome showed absorption peaks at 276, 411 and 523 nm in the oxidized form and peaks at 315, 417, 523 and 552 nm in the reduced form. The molecular weight of the cytochrome was estimated to be 13,800 on the basis of the amino acid composition and heme content, and 14,000 from SDS-polyacrylamide gel electrophoresis analysis. Its midpoint redox potential at pH 7.0 was determined to be +0.36 V. The N-terminal amino acid sequence of the cytochrome was determined as follows: A-G-G-A-G-G-P-A-P-Y-R-I-S-? D-?-M-V-?-S-G-M-P-G-. Ferrocytochrome c-552 was oxidized by the membrane fraction of T. ferrooxidans, and the oxidation rate was more rapid at pH 3.0 than at pH 6.5. Ferricytochrome c-552 was reduced by Fe(II)-cytochrome c oxidoreductase with Fe2+ at pH 3.5, while horse ferricytochrome c was not reduced by the enzyme under the same reaction conditions. PMID- 2551388 TI - Calcium-induced localization of calcium-activated neutral proteinase on plasma membranes. AB - The location of calcium-activated neutral proteinase (CANP) was determined in human erythrocytes by crosslinking CANP to co-localizing proteins using a photolabeling bifunctional reagent, 4,4'-dithiobisphenylazide (DTBPA). The crosslinked products were selectively isolated by immunoprecipitation with a polyclonal anti-CANP antibody and analyzed by SDS-polyacrylamide gel electrophoresis after cleavage of the crosslinkage. In the calcium-free incubation medium the main proteins crosslinked with CANP were cytosolic proteins such as hemoglobin. In the presence of calcium ions, on the other hand, membrane skeletal proteins such as spectrin, band 4.1, 4.2 and 6 proteins as well as band 3 were crosslinked with CANP. Addition of calcium ionophore further increased the amount of crosslinked membrane proteins. These results suggest that in the absence of calcium ions CANP exists diffusely in the cytoplasm and is crosslinked with cytoplasmic hemoglobin nonspecifically while in the presence of calcium ions CANP associated with membrane where it is crosslinked specifically with the lining proteins. Thus it is demonstrated biochemically that the localization of CANP is dynamic depending on the presence of calcium ions. PMID- 2551387 TI - Monomeric bacteriochlorophyll is required for the triplet energy transfer between the primary donor and the carotenoid in photosynthetic bacterial reaction centers. AB - Reaction centers from the carotenoidless mutant Rb. sphaeroides R26 were treated with sodium borohydride which is known to remove one of the accessory monomeric bacteriochlorophylls (BB). Subsequently, the carotenoid, spheroidene, was incorporated into the modified reaction centers. It is demonstrated by optical absorption and circular dichroism experiments that spheroidene, reconstituted into the sodium borohydride-treated Rb. sphaeroides R26 reaction centers, is bound in a single site, in the same environment and with the same structure as spheroidene reconstituted into untreated (native) Rb. sphaeroides R26 reaction centers. Transient optical and electron spin resonance spectroscopic data indicate that unless the accessory BB is present, the primary donor-to-carotenoid triplet energy transfer reaction is inhibited. These observations provide direct evidence for the involvement of the accessory BB in the triplet energy transfer pathway. PMID- 2551389 TI - Identification of Na+/K+-ATPase inhibitors in bovine plasma as fatty acids and hydrocarbons. AB - A preparative purification of endogenous inhibitors of the Na+/K+-ATPase has been carried out from bovine blood. Dried plasma was deproteinized, hexane-extracted and desalted, followed by further purification through a series of reverse-phase HPLC fractionations. Fractions active in inhibiting Na+/K+-ATPase activity and displacing ouabain were collected and purified further. By comparison with ouabain, the final extract was found to have a steeper concentration-effect curve in the inhibition of Na+/K+-ATPase. In displacement of [3H]ouabain, the extract had again a steeper concentration-effect curve than does ouabain, and in addition it enhanced ouabain binding at high dilutions. These properties are indicative of nonspecific interactions with the Na+/K+-ATPase. The active fraction was identified by TLC, HPLC, NMR, GLC and GC-MS, to be a mixture of three unesterified fatty acids, mainly oleic acid (72% of the total) and three saturated hydrocarbons. The assignment of structures was corroborated by comparison with authentic samples. PMID- 2551390 TI - Studies of the action of symmetrical and mixed disulphide thiol-modifying reagents on the esterase activity of cytoplasmic aldehyde dehydrogenase. AB - The effect of various thiol-modifying reagents on the esterase activity of sheep liver cytoplasmic aldehyde dehydrogenase is reported here. Both symmetrical reagents (disulfiram, 2,2'- and 4,4'-dithiodipyridines) and unsymmetrical reagents (methyl diethylthiocarbamyl disulphide, methyl 2- and 4-pyridyl disulphides) were investigated. The results suggest that all the modifiers react to varying extents with a pair of enzymic thiol groups ('A' and 'B'), and that the more specifically group 'A' is modified, the more the enzyme is inactivated. This supports the idea that group 'A' may be the essential nucleophile in the reaction catalysed by aldehyde dehydrogenase. Modification of group 'B' may or may not reduce the esterase activity depending on the nature of the label introduced. The results of the present experiments and of previous similar experiments concerning the dehydrogenase activity of the enzyme are consistent with the proposal that a common active site is responsible for both esterase and dehydrogenase activities. PMID- 2551391 TI - Isolation and characterization of biologically active murine interleukin-1 alpha derived from expression of a synthetic gene in Escherichia coli. AB - A murine interleukin-1 alpha (mIL-1 alpha) gene coding for amino acids 115 to 270 of the precursor protein (Lomedico, P.T., Gubler, U., Hellmann, C.P., Dukovich, M., Giri, J.G., Pan, Y.E., Collier, K., Semionow, R., Chua, A.O. and Mizel, S.B. (1984) Nature 312, 458-462) was chemically synthesized and expressed in Escherichia coli. mIL-1 alpha, in the form of insoluble inclusion bodies, accounted for approx. 30% of total cellular protein produced by the recombinant strain. A simple isolation protocol was developed in which inclusion body material was first solubilized in 3 M guanidine hydrochloride, and the mIL-1 alpha was then simultaneously purified and allowed to fold to its active conformation by dialysis against distilled water. This procedure yielded pure, biologically active mIL-1 alpha with 41% recovery of the mIL-1 alpha present in the guanidine hydrochloride extract. The purified preparation had the expected amino acid composition, a molar absorptivity of 28,200 M-1.cm-1 and a pI of 5.2. No methionyl-mIL-1 alpha was detected by N-terminal sequence analysis, and the endotoxin level was less than 10 pg per micrograms of mIL-1 alpha. The specific biological activity was 3.10(7) units/mg in a co-mitogenic thymocyte proliferation assay. In addition to full-length mIL-1 alpha, the preparation contained N-terminally truncated mIL-1 alpha species (mainly des-4 and des-6 amino acid forms). The truncated species were isolated and found to have the same biological activity as the complete polypeptide. Thus, the active fragment of mIL 1 alpha appears to consist of a proteinase-sensitive N-terminal region which is not essential for activity, and a proteinase-resistant core which harbors the essential determinants of its cytokine function. PMID- 2551392 TI - Modeling the olfactory bulb and its neural oscillatory processings. AB - The olfactory bulb of mammals aids in the discrimination of odors. A mathematical model based on the bulbar anatomy and electrophysiology is described. Simulations of the highly non-linear model produce a 35-60 Hz modulated activity which is coherent across the bulb. The decision states (for the odor information) in this system can be thought of as stable cycles, rather than point stable states typical of simpler neuro-computing models. Analysis shows that a group of coupled non-linear oscillators are responsible for the oscillatory activities. The output oscillation pattern of the bulb is determined by the odor input. The model provides a framework in which to understand the transform between odor input and the bulbar output to olfactory cortex. There is significant correspondence between the model behavior and observed electrophysiology. PMID- 2551393 TI - [NADH- and NADPH-dependent formation of superoxide radicals in liver nuclei]. AB - A method for the quantitation of the superoxide radical generation rate (V) in murine liver nuclei by the oxidation of 1-hydroxy-2,2,6,6-tetramethyl-4-oxo piperidine O2-. radicals with the formation of a stable nitroxyl radical recorded by the EPR method, has been developed. It was shown that NADP- and NADPH dependent superoxide radical generation is suppressed by superoxide dismutase (approximately by 90%). The Km values for NADH and NADPH are 1.5 x 10(-6) and 4.4 x 10(-7) M, respectively; the maximal rate (0.2 nmol.min-1.mg protein-1) is equal for both substrates. Cyanide (greater than 2 mM) causes a practically complete inhibition of the O2-. generation by both substrates. It is suggested that there exists a single readily autooxidized site of O2-. generation by both substrates for NADH- and NADPH-dependent site of the electron transport chain in nuclear membranes. PMID- 2551394 TI - [The role of the substrate structure in the function of Na,K-ATPase]. AB - The mechanism of Na,K-ATPase function is reviewed. The peculiarities of hydrolysis of various substrates are described. The experimental results testify to the effect of substrate structure on the E2----E1 transition, rate of Na+ transport, K-dependent phosphatase activation and the quaternary structure of Na,K-ATPase. A conclusion is drawn that the proton-acceptor properties of the substrate play a role in the regulation of ion transport by Na,K-ATPase. PMID- 2551395 TI - [Biochemical features of platelet alpha2-adrenergic receptors and their connection with an increase in concentration of intracellular Ca2+]. AB - Epinephrine (E) and norepinephrine (NE) alone did not increase free intracellular Ca2+ ([Ca2+]i) in human platelets loaded with Quin-2 or Fura-2; however, they did potentiate the effects of vasopressin (VP), serotonin (S) and platelet activating factor (PAF). The synergism in [Ca2+]i increase was also obtained in the presence of VP together with PAF, S with PAF as well as VP with S. The effect of E or NE was blocked by yohimbine and phentolamine. Prazosin was less effective, while propranolol had no effect at all. Clonidine did not potentiate the effects of VP, S or PAF on [Ca2+]i; however, it did block the potentiation induced by E or NE. E potentiated the VP-induced 45Ca2+ uptake as well as VP-stimulated inositol 1,4,5 trisphosphate (IP3) formation. E alone did not change significantly the level of IP3 in platelets, nor did it influence the cyclic AMP level. The experimental results suggest that both Ca2+ influx and polyphosphoinositide breakdown underlie the mechanism of potentiation. PMID- 2551396 TI - [Introduction of regulatory subunits of cAMP-dependent protein kinase type II in ZTZ cells with the use of erythrocyte ghosts]. AB - The dynamics of distribution of the regulatory subunit of cAMP-dependent protein kinase II following protein injection into 3T3 cells was studied. The cAMP binding component of protein kinase was injected into the cells, using erythrocyte ghosts. The conditions for protein encapsulation into erythrocyte ghosts were elaborated. The optimal detergent concentrations, incubation time and conditions of vesicular closure following protein injection were selected. The above method provides for a high (50-55%) yield of the erythrocyte ghost encapsulated protein with a minimum loss of enzymatic activity. Fusion of erythrocyte ghosts containing the labeled protein with 3T3 cells was carried out. Using the cytoradiography technique, the dynamics of distribution of the radiolabeled regulatory subunit within the cell was analyzed. It was demonstrated that after the regulatory subunit has reached the cytoplasm, the protein is translocated into the nucleus and is pooled there is the vicinity of the nucleoli. PMID- 2551397 TI - A.E. Bennett award paper. Experimental approaches to human stress research: assessment of neurobiological mechanisms of stress in volunteers and psychiatric patients. AB - This article presents a series of experiments that involves the development of three novel strategies for human stress research and the utilization of these strategies to examine neurobehavioral processes of stress in healthy volunteers, schizophrenia, and affective illness. The first strategy involved intravenous 2 deoxy-D-glucose (2DG) administration, a glucoprivic stressor. We found that glucoprivic stress results in dissociation of hypothalamus-pituitary-adrenal (HPA), adrenomedullary, and sympathoneural activity. In addition, glucoprivic stress in neuroleptic-treated schizophrenic patients caused heightened dopamine activity, as reflected by increased plasma homovanillic acid (HVA) levels and decreased adaptive responses as assessed by decreased food consumption following 2DG administration. These data suggest that neuroleptics do not prevent stress related increases in dopamine activity and that schizophrenia may be associated with abnormalities in the stress response. The second strategy assessed effects of uncontrollable and identical amounts of controllable stress in volunteers and depressed patients. In volunteers, it was found that uncontrollable in comparison to controllable stress results in specific behavioral and neuroendocrine alterations. Moreover, uncontrollable stress exposure in depressed patients in comparison to volunteers produced greater alterations in behavioral ratings and plasma cortisol levels and that the uncontrollable stress related increases in helplessness ratings and cortisol levels were significantly correlated. These data suggest that depressed patients may have increased sensitivity to uncontrollable stress and that there may be an important interrelationship between the cognitive deficits of depression and the heightened HPA axis activity observed in these patients. Lastly, we used a naturalistic strategy to examine mechanisms relating childhood parental loss and the development of adult affective illness and found that among subjects with early parental loss histories, those who developed adult psychiatric illness had increased resting plasma levels of cortisol and beta-endorphin (ir) as compared with subjects with early loss and no adult history of psychiatric illness. Moreover, increased HPA activity in adulthood was significantly related to poor childhood adjustment to parental loss. The implications of the results of these studies are discussed. PMID- 2551398 TI - Biological correlates of lactate sensitivity in panic disorder. AB - In this pilot study, 9/9 patients with panic disorder experienced a lactate induced panic attack as compared with 0/9 controls. Baseline measurements were significant for higher anxiety self-ratings, higher heart and respiratory rates, elevated potassium, and lower lymphocyte 3H-dihydroalprenolol (DHA) binding in the patient group. Spielberger State anxiety scores correlated with baseline mean heart rate, and Spielberger Trait anxiety scores with lymphocyte DHA binding. The lactate infusion was not found to differentially affect any physiological or biochemical measures in the two groups, though heart rate surges occurred in most patients. Intravenous propranolol reduced the panic to a negligible degree, whereas intravenous diazepam was quite effective. Neurobiological implications are discussed, and the contradictory biological findings in the lactate literature are reviewed. PMID- 2551399 TI - Changes in platelet 3H-imipramine binding with chronic imipramine treatment are not state-dependent. AB - One month of imipramine treatment increased both the Kd and Bmax of platelet 3H imipramine binding in 11 endogenous unipolar depressed patients. Continued treatment (13 weeks) of 5 patients subsequently lowered the Bmax values of 2 patients who had initially shown the largest increases, so that binding was no longer significantly elevated after 13 weeks. The observed changes in Kd but not in Bmax, could be explained by the carryover of tightly bound drug to the binding assay, although neither of the measures were correlated with plasma imipramine levels. Posttreatment Bmax (4 weeks) values were inversely related to plasma cortisol levels, although a weak but positive correlation was found before treatment. No significant change was found in plasma cortisol with treatment. Clinical responses were not related to cortisol or Bmax changes, although optimal improvement was associated with extreme values (high and low) of pretreatment Bmax. The present results, obtained with imipramine, and similar results obtained after nortriptyline and electroconvulsive shock by others, suggest that at least some antidepressants may induce transient changes in the Bmax of platelet binding that are independent of affective state. PMID- 2551400 TI - Seasonal rhythm of platelet 3H-imipramine binding in normal controls. AB - The densities of platelet 3H-imipramine sites were determined by repetitive measures of 11 normal controls over the course of 1 year. A significant seasonal variation was found, with a circannual peak on February 17 and a nadir on August 18. The estimated amplitude of this rhythm was +/- 599.54 fmol/mg, which fluctuated about a yearly mean of 2647.5 fmol/mg. The present results underscore the importance of including seasonally matched controls in the evaluation of potential patient differences in platelet binding. PMID- 2551401 TI - Erythrocyte membrane Na-K ATPase activity in affective disorder. PMID- 2551402 TI - The National Institute of Mental Health: prospects and promises. PMID- 2551403 TI - Pavlovian conditioning of pain regulation: insights from pharmacological conditioning with morphine and naloxone. PMID- 2551404 TI - Integer-spin electron paramagnetic resonance of iron proteins. AB - A quantitative interpretation is presented for EPR spectra from integer-spin metal centers having large zero-field splittings. Integer-spin, or non-Kramers, centers are common in metalloproteins and many give EPR signals, but a quantitative understanding has been lacking until now. Heterogeneity of the metal's local environment will result in a significant spread in zero-field splittings and in broadened EPR signals. Using the spin Hamiltonian Hs = S.D.S + beta S.g.B and some simple assumptions about the nature of the zero-field parameter distributions, a lineshape model was devised which allows accurate simulation of single crystal and frozen solution spectra. The model was tested on single crystals of magnetically dilute ferrous fluosilicate. Data and analyses from proteins and active-site models are presented with the microwave field B1 either parallel or perpendicular to B. Quantitative agreement of observed and predicted signal intensities is found for the two B1 orientations. Methods of spin quantitation are given and are shown to predict an unknown concentration relative to a standard with known concentration. The fact that the standard may be either a non-Kramers or a Kramers center is further proof of the model's validity. The magnitude of the splitting in zero magnetic field is of critical importance; it affects not only the chance of signal observation, but also the quantitation accuracy. Experiments taken at microwave frequencies of 9 and 35 GHz demonstrate the need for high-frequency data as only a fraction of the molecules give signals at 9 GHz. PMID- 2551405 TI - Microsecond rotational motion of spin-labeled myosin heads during isometric muscle contraction. Saturation transfer electron paramagnetic resonance. AB - We have used saturation transfer electron paramagnetic resonance (ST-EPR) to detect the microsecond rotational motions of spin-labeled myosin heads in bundles of skinned muscle fibers, under conditions of rigor, relaxation, and isometric contraction. Experiments were performed on fiber bundles perfused continuously with an ATP-regenerating system. Conditions were identical to those we have used in previous studies of myosin head orientation, except that the fibers were perpendicular to the magnetic field, making the spectra primarily sensitive to rotational motion rather than to the orientational distribution. In rigor, the high intensity of the ST-EPR signal indicates the absence of microsecond rotational motion, showing that heads are all rigidly bound to actin. However, in both relaxation and contraction, considerable microsecond rotational motion is observed, implying that the previously reported orientational disorder under these conditions is dynamic, not static, on the microsecond time scale. The behavior in relaxation is essentially the same as that observed when myosin heads are detached from actin in the absence of ATP (Barnett and Thomas, 1984), corresponding to an effective rotational correlation time of approximately 10 microseconds. Slightly less mobility is observed during contraction. One possible interpretation is that in contraction all heads have the same mobility, corresponding to a correlation time of approximately 25 microseconds. Alternatively, more than one motional population may be present. For example, assuming that the spectrum in contraction is a linear combination of those in relaxation (mobile) and rigor (immobile), we obtained a good fit with a mole fraction of 78-88% of the heads in the mobile state. These results are consistent with previous STEPR studies on contracting myofibrils(Thomas et al., 1980). Thus most myosin heads undergo microsecond rotational motions most of the time during isometric contraction, at least in the probed region of the myosin head.These motions could arise primarily from the free rotations of heads detached from actin. However, if most of these heads are attached to actin during contraction, as suggested by stiffness measurements, this result provides support for the hypothesis that sub-millisecond rotational motions of actin-attached myosin heads play an important role in force generation. PMID- 2551406 TI - Reconstruction of the probe angular distribution from a series of electron spin resonance spectra of tilted oriented samples. AB - Model-independent methods for the reconstruction of the nitroxide spin probe angular distribution of labeled oriented biological assemblies from electron spin resonance (ESR) spectra were investigated. We found that accurate probe angular distribution information could be obtained from the simultaneous consideration of a series of ESR spectra originating from a sample at differing tilt angles relative to the Zeeman magnetic field. Using simulated tilt series data sets, we developed a consistent criteria for judging the reliability of the simulated fit to the data as a function of the free spectral parameters and thereby have increased the significance of the model-independent reconstruction of the probe angular distribution derived from the fit. We have also enhanced the angular resolution measurable with the model-independent methodology by increasing the rank of the order parameters that we can reliably deduce from a spectrum. This enhancement allows us to accurately deduce higher resolution features of the spin probe distribution. Finally we investigated the usefulness of fitting the tilt series data in multiple data sets such that tilt series data from many identical sample preparations are fitted simultaneously. This method proved to be useful in rapidly reducing a large amount of data by eliminating any redundant computations in the application of the enhanced model-independent analysis to identical sets of tilt series data. We applied the methodology developed here to ESR spectra from probe labeled muscle fibers to study the orientation of myosin cross-bridges in fibers. This application is described in the accompanying paper. PMID- 2551407 TI - Myosin cross-bridge orientation in rigor and in the presence of nucleotide studied by electron spin resonance. AB - The tilt series electron spin resonance (ESR) spectrum from muscle fibers decorated with spin labeled myosin subfragment 1 (S1) was measured from fibers in rigor and in the presence of MgADP. ESR spectra were measured at low amplitude modulation of the static magnetic field to insure that a minimum of spectral lineshape distortion occurs. Ten tilt series ESR data sets were fitted simultaneously by the model-independent methodology described in the accompanying paper (Burghardt, T. P., and A. R. French, 1989. Biophys. J. 56:525-534). By this method the average and standard error in the mean of order parameters for the probe angular distribution were calculated for the two states of the fiber investigated. The average order parameters were used to reconstruct the probe angular distribution in two dimensions, one angular dimension corresponding to a polar angle measured relative to the fiber axis, and the other a torsional angular degree of freedom of the probe. We find that the probe angular distributions for the rigor and MgADP states of the fiber differ such that the rigor distribution is broader and shifted relative to the distribution in the presence of MgADP. The shape of the rigor distribution suggests the presence of two probe orientations, one similar to that in the presence of MgADP, and another at a different orientation. The shape of the distribution in the presence of MgADP suggests that the binding of the nucleotide to the rigor cross-bridge shifts the spin population into a more homogeneous one by causing a cross-bridge rotation. PMID- 2551408 TI - Ionic diffusion in voltage-clamped isolated cardiac myocytes. Implications for Na,K-pump studies. AB - The whole-cell voltage-clamp technique employing electrolyte-filled micro-pipette suction electrodes is widely used to investigate questions requiring an electrophysiological approach. With this technique, the ionic composition of the cytosol is assumed to be strongly influenced (as result of diffusion) by the ionic composition of the solution contained in the electrode. If this assumption is valid for isolated cardiac myocytes, the technique would be particularly powerful for studying the dependence of their Na,K-pump on the intracellular [Na+]. However, the relationship between the concentrations of ions in the solution filling the electrode and those in the cytosol has not been established. The relationship was investigated to determine in particular whether the [Na+] at the intracellular cation ligand binding sites for the Na-pump ([ Na+]ps) can be set and clamped by [Na+] in the pipette electrode ([ Na+]pip). If [Na+]pip can set and clamp [Na+]ps, this would provide a means for defining the dependence of the Na,K-pump on intracellular [Na+]. The relationship between [Na+]pip and [Na+]ps was analyzed using two approaches. First, a mathematical model of three dimensional ionic diffusion within a whole-cell patch-clamped myocyte was developed and the effects of experimental parameters on mean [Na+]ps were investigated. When typical experimental values were simulated, the time course to achieve steady state mean [Na+]ps was found to be most sensitive to variations in electrode pore size, cell length and the Na+ pumping rate, but at steady state, mean [Na+]ps varies from [Na+]pip by 5% or less depending on pump rate. Second, to provide experimental support for the validity of the simulations, isolated ventricular myocytes were voltage-clamped and the reversal potential for the Na current was determined in order to estimate steady state intracellular [Na+]. The results of the mathematical and experimental analyses suggest that steady state [Na+]ps can be regulated by the [Na+] in suction pipette electrodes. These findings, while also having a broader significance, indicate for isolated cardiac myocytes that whole-cell suction micro-electrodes can provide a means to assess the dependence of the Na,K-pump on [Na+]ps. PMID- 2551410 TI - Human lymphocyte and myocardial beta-adrenoceptors: up and down regulation. AB - Radioreceptor assays have extended the knowledge of beta-adrenoceptor regulation. To evaluate the usefulness of this method in cardiovascular pharmacology, this study was designed to determine: (1) the correlation between beta-adrenoceptor density in human lymphocytes and myocardial cell membranes. This was achieved by using simultaneously harvested left auricles and whole blood samples in 10 patients scheduled to undergo cardiothoracic surgery. The correlation was found to be linear (r = 0.65; P less than 0.05; N = 10); (2) the changes in lymphocyte receptor density and affinity in heart failure. Lymphocytes were harvested from 6 healthy volunteers, 8 patients with moderate heart failure (NYHA class I or II) and 8 patients with severe heart failure (NYHA class III or IV). Mean densities observed were 75.6 +/- 11, 46.3 +/- 18 and 26.4 +/- 5.9 fmol/mg protein, respectively, and dissociation constants were 62.8 +/- 16; 67.8 +/- 14, and 46 +/ 26 pM. The number of receptors fell significantly from one heart failure class to that immediately above it (P less than 0.01; N = 22); (3) the beta adrenoceptor regulatory properties of a class I antiarrhythmic drug, propafenone, the chemical structure of which is similar to that of the beta-blocking drug propranolol. Five patients needing antiarrhythmic treatment were given 10 d of oral propafenone treatment (450-900 mg/day). Mean adrenoceptor densities (Bmax) were 22.7 +/- 9 and 43.7 +/- 9 fmol/mg protein, and dissociation constants (KD) values were 24.7 +/- 20 and 27 +/- 18 pM respectively, before and after 10 d of chronic treatment. The number of receptors increased significantly after 10 d of treatment (P less than 0.01; N = 5).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551409 TI - Turnover of epidermal growth factor binding sites in mouse mammary epithelial cells. AB - An in vitro method of studying epidermal growth factor (EGF) receptors in mouse mammary epithelial cells in serum-free collagen gel culture has been developed. Binding of EGF averaged 108 +/- 19 fmol/mg DNA in cells isolated from freshly dissociated virgin mammary tissue. Initial binding values were maintained in cells cultured in the presence of 0.1 ng/ml EGF, but decreased in either 0 ng/ml or 10 ng/ml EGF. Addition of either chloroquine (100 microM) or ammonium chloride (10 mM) to the culture medium increased receptor levels 10-fold. Cycloheximide (0.1 microgram/ml), ouabain (3 mM), and actinomycin D (5 x 10(-2) micrograms/ml) each decreased receptor levels, in some cases by as much as 80%. Both methylamine (10 mM) and dinitrophenol (0.1 mM) had no significant effect. These studies suggest that the net level of EGF receptors in these target cells is the result of an equilibrium between synthesis and degradation. The difference between the effects of the compounds tested on either receptor degradation or synthesis in comparison to cell growth, may be indicative that receptor degradation is not linked to cell proliferation. PMID- 2551411 TI - Short-term effects of omega-3 fatty acids on exercise stress test parameters, angina and lipoproteins. AB - The short-term effects of omega-3 fatty acids on exercise test parameters in patients with known coronary artery disease are poorly defined. Therefore, we administered a preparation containing 5.4 g of eicosapentaenoic acid and 3.6 g docosahexaenoic acid to 8 patients in a random order, double-blind, crossover design. The patients were given either fish oil or a vegetable oil control for 6 wk and then crossed over to the other treatment for a second 6-wk period, after a 2-wk wash-out interval. Exercise test parameters, angina frequency, and lipoprotein composition were measured at baseline and at the end of each trial phase. No significant changes were found in resting or peak exercise heart rate or double product, exercise duration or ischemic electrocardiographic changes at peak exercise. Angina frequency and sublingual nitroglycerin usage did not change during either the fish oil or the control oil phase. Plasma triglyceride levels decreased significantly with fish oil treatment and HDL cholesterol levels rose slightly. Total and LDL cholesterol levels were unchanged. We conclude that relatively high dose, short-term administration of omega-3 fatty acids does not alter exercise test parameters. PMID- 2551412 TI - [Dependence of the degree of cardiac stress damages on the change in the endogenous beta-endorphin level during preliminary adaptation]. AB - It was found that simulation of stress of the white rats increases the myocardial uptake of Tc-99m-pyrophosphate (Tc-pyp). The before-adapted group of animals had a sufficiently lower content of Cc-PYP in myocardium, than the stress-control group. At that time, the before-adaptation led to a low rise in the plasma beta endorphin and to a significant rise in its quantity. The authors discuss the role of endogenous beta-endorphin in the mechanisms of the protecting effect of adaptation. PMID- 2551413 TI - [Effect of opiate receptor agonists on the course of hemorrhagic shock in rats]. AB - The experiments have been performed on 93 male rats, weighing 200-250 g. In acute blood loss various arterial pressure (AP) changes have been demonstrated--the marked hypertension is being changed by gradual AP increase. The injection of m receptors' agonist DAGO prevents systolic and diastolic AP increase, agonist DADL prevents diastolic AP increase in acute momentary blood loss. In gradual blood loss DAGO (more than DADL) slows down both the decrease and the subsequent AP increase in rats. DAGO is determined to decrease, and DADL--to increase the minute blood volume. The mechanisms of opioids' action and their significance in pathogenesis of hemodynamic disturbances in shock are being discussed. PMID- 2551414 TI - [Effect of the Ca ionophore A-23187 on the plasmatic and mitochondrial potentials of the brain synaptosomes in rats: fluorescence measurements]. AB - The applicability of the potential-sensitive dye diS-C3-(5) for the study of A23187 + Ca2+ induced plasma membrane hyperpolarization was tested in rat brain synaptosomes. An appropriate dye synaptosome ratio was chosen for the fluorescence titration dye in Ca-free Krebs-Ringer solution. The fluorescence intensity of the probe was increased upon the addition of Ca2+ (1 microM) to the synaptosomes in the presence of A23187 (1 microM). The effect of Ca2+ + A23187 persisted in a Na+-free medium or when Na+ channels were inhibited by tetrodotoxin as well as in high K+-depolarized synaptosomes (75 microM KCl). In the presence of oligomycin or a protonophore (1 microM) the effect of Ca2+ + A23187 was suppressed. This suggests that the A23187-induced fluorescence increase is due to a depolarization of intrasynaptosomal mitochondria. Therefore, the use of the dye diS-C3-(5) for the study of Ca-induced hyperpolarization does not seem to be feasible unless a quantitative model of changes in fluorescence related to the plasma and mitochondrial membrane potentials is elaborated. PMID- 2551415 TI - [The generation of active microbicidal forms of oxygen by leukocytes transiting the vascular bed of the lungs]. AB - In experiments on dogs using the chemiluminescent method and nitroblue-tetrazole reaction the authors found out that leucocytes while passing through the pulmonary vessels bed, in contrast to the spleen increase the generation of active microbicidal forms of oxygen. Due to this fact we suppose that the lungs may take part in the formation of free radical status and phagocytic antiinfectious defence of the organism. PMID- 2551416 TI - [Anticonvulsive properties of peptide ACTH4-7 pro-gly-pro detected in amygdaloid kindling and audiogenic epilepsy in rats]. AB - Effects of the ACTH4-7 pro-gly-pro, calcium valproate ("Germed", DDR) and nembutal on kindling preparation and audiogenic epilepsy were investigated. Development of after-discharges in response to repeated amygdaloid electrical stimulation was assessed in normal rats and in rats susceptible to audiogenic epilepsy (KM line of rats). ACTH4-7 pro-gly-pro had an anticonvulsant profile. ACTH4-7 pro-gly-pro decreased seizure threshold in the audiogenic epilepsy test, but did not prevent the motor convulsions. PMID- 2551417 TI - gamma-Aminobutyric acid-A receptor-mediated suppression of 5-hydroxytryptamine induced guinea-pig basilar artery smooth muscle contractility. AB - The mechanism of gamma-aminobutyric acid (GABA)-induced suppression of 5 hydroxytryptamine (5HT)-induced contractility of cerebral blood vessels was studied in single smooth muscle cells isolated from the guinea-pig basilar artery. GABA reduced 5HT-induced contraction of single smooth muscle cells, and the effect of GABA was mimicked by muscimol, but not baclofen. The response of muscimol was antagonized by bicuculline, thereby indicating that GABAA receptors exist on the smooth muscle of the basilar artery. Since GABA did not change the contraction induced by the addition of Ca2+ to the Ca2+-free medium in the presence of high K+, it is unlikely that GABA inhibits the influx of extracellular Ca2+. The caffeine-induced contraction in the Ca2+-free medium was reduced by GABA, and the effect of GABA was not obtained by treatment with furosemide and in the Cl- -free medium. These results indicate that GABA acts on the GABAA receptor located on smooth muscle cells and reduces the contractility of the basilar artery by suppression of the mobilization of intracellular Ca2+. PMID- 2551418 TI - Methionine-enkephalin stimulates in vitro proliferation of human peripheral lymphocytes via delta-opioid receptors. AB - The influence of methionine-enkephalin (Met-Enk) on in vitro proliferation of human peripheral lymphocytes was investigated. Met-Enk, in the concentration range 10(-12) to 10(-4) M enhanced the proliferative response to suboptimal concentrations of concanavalin A of human peripheral lymphocytes and to cells in the absence of mitogen. The response to Met-Enk in the absence of mitogen was not influenced by the presence of fetal calf serum: similar levels of enhancement were seen in cultures supplemented with 10% autologous serum. Enhancement of proliferation was blocked in a concentration-dependent manner by both naloxone and the delta specific antagonist ICI-174864. The sensitivity to the antagonistic influence of ICI-174864 suggests strongly that the stimulatory influence of Met Enk on human lymphocyte proliferation in the absence of mitogen is mediated via the delta-opioid receptor. PMID- 2551419 TI - Characterization of virus infected cell cultures by pyrolysis/direct chemical ionization mass spectrometry. AB - The supernatants of Vero cell cultures after infection with a herpes simplex virus or a poliomyelitis virus as well as a blank were analysed by pyrolysis/direct chemical ionization mass spectrometry (Py/DCI MS). Informative pyrogrammes were obtained and used for characterization of viral proteins by applying pattern recognition methods. Differentiation of viral proteins was evaluated by analysing 'blind' samples. Herpes viruses could be classified correctly but the observed differences between the blank and the polio virus supernatants were too small for reliable classification of the polio viruses. Purification of the samples seems to be a prerequisite for further studies. The potential value of Py/DCI MS as a rapid non-invasive diagnostic method for viral meningoencephalitis is stressed. PMID- 2551420 TI - Surgical considerations and planning for acetabular and femoral deficiencies in revision hip replacement. AB - Cementless revision total hip arthroplasty provides a method for augmenting and restoring the remaining bone stock in a failed total hip arthroplasty. Careful preoperative planning, precise intraoperative technique and an adequate prosthetic inventory are required. Classification systems for bone deficiencies aid in planning grafting and prosthetic needs and in reviewing postoperative results. New cementless technology should provide even more satisfying results. PMID- 2551421 TI - The use of bone grafts for support and fixation in revision total hip arthroplasty. AB - An increasing number of orthopaedic surgical cases involve revision total hip replacement, not only first time revisions, but often second, third, or more. With each revision, the number of good results tends to decline. The authors present a technique that makes use of bone grafts to support and strengthen the fixation of hip prostheses in total joint replacement surgery. PMID- 2551422 TI - Osteonecrosis of both medial tibial condyles following short-term steroid therapy. A case report. AB - The authors present an unusual case of steroid-induced osteonecrosis in both medial tibial condyles and in no other joint or skeletal site. PMID- 2551423 TI - Nocardia asteroides infection of an Austin-Moore hemiarthroplasty in a nonimmunocompromised host. A case report. AB - A case of Nocardia asteroides infection of a hip prosthesis in a nonimmunocompromised patient is presented. The infection developed soon after the operation, and did not respond to empiric treatment by multiple antimicrobial drugs. Reoperation and removal of the prosthesis was necessary. The authors found no previous cases of nocardiosis complicating arthroplasty reported in the literature. PMID- 2551424 TI - Management of an infected total knee arthroplasty. AB - Infection following total knee arthroplasty can be one of the most challenging problems in orthopaedic surgery. This article discusses the pertinent clinical factors to be considered and the treatment options in the management of patients with infection following total knee replacement. PMID- 2551425 TI - Avoiding knee skin sloughs. AB - To avoid skin sloughs about the knee following knee arthroplasty, the principles that need to be emphasized are the blood supply in the area so that appropriate incisions can be designed; careful operative technique to avoid hematomas; appropriate use of drains and dressings; and postoperative care that monitors increased range-of-motion as it affects the wound. Should secondary coverage be needed, the options include secondary closure, skin graft, muscle or myocutaneous flaps, and on rare occasions, a free tissue transfer. In this way, the plastic surgeon can assist the orthopaedist by providing the appropriate soft tissue coverage in difficult knee surgical situations. PMID- 2551426 TI - Metal-backed patellar component failure and metallic synovitis in total knee arthroplasty. A case report. AB - A case of a dissociation of a metal-backed patellar component with a resultant metallic synovitis is presented. The mechanism of failure is discussed and an analysis of patellar component design is reviewed. PMID- 2551427 TI - A critical review. Heterotopic ossification in total hip replacement. AB - Heterotopic ossification (HO) status post total hip arthroplasty is a relatively common phenomenon with clinical significance in approximately 5% of all cases. Risk factors appear to include males with osteoarthritis, particularly with marked osteophyte formation, and those with ankylosing spondylitis or diffuse idiopathic spinal hyperostosis. Previous hip surgery, or previous ectopic bone in the same or contralateral hip are definite predisposing factors. Although meticulous surgical technique is critical in any operation, the suggestions that carelessness in dissection or tissue handling, or inadequate hemostasis or debridement of devitalized tissues or of bony debris can cause HO are unproved. Similarly, there is no solid evidence that the surgical approach, prosthesis type, use of trochanteric osteotomy, or the presence of cement influence the incidence of HO. Whether postoperative complications such as infection, dislocation, or hematoma are causally related is speculative; and the role of alkaline phosphatase in predicting those at risk remains controversial. Despite the number of studies designed to elucidate risk factors, critical analysis suggests that this question remains largely unanswered and that there is a need for well-designed, prospective, controlled studies to determine which hip arthroplasty patients are at risk. Treatment of established HO depends upon recognizing the "maturity" of the ectopic bone, which can best be determined by serial scans but is approximately one year postop. Excision followed by prompt initiation of radiotherapy or of one of several reported nonsteroidal anti inflammatory drug protocols will produce successful results in a majority of cases. Prophylaxis depends upon recognizing those at significant risk and initiating the appropriate protocol within the first few postoperative days. PMID- 2551428 TI - Calcinosis circumscripta: indications for surgery. AB - Five patients with calcinosis circumscripta were treated, four of them surgically. The indications for surgery included painful lumps, recurrent infection of the calcified masses, secreting ulcers, functional disorders, severe esthetic problems, and pressure on nerves. One patient showed permanent improvement; four gained temporary relief. PMID- 2551429 TI - Comparison of sensory latencies of the median nerve at the carpal tunnel among juveniles and adults. AB - Five hundred and ninety-three juveniles and adults were studied to determine if there was an age-related trend in the occurrence of sensory nerve conduction latencies of the median nerve at the carpal tunnel. Latencies were found to increase with increasing age in both the dominant and nondominant hands of the subjects. A significant and positive correlation was found between the latencies of both hands, indicating that an increased value in one hand is likely to be associated with an increased value in the other. Among the adults, the latencies were found to be consistently greater for the dominant hand. PMID- 2551430 TI - Immunohistochemical localization of type II collagen in cartilage-forming tumors. AB - The presence and distribution of type II collagen was studied in 36 cartilage and cartilage-related tumors, including five osteosarcomas and one chordoma. A monoclonal antibody prepared from chicken type II collagen was used with paraffin sections, employing the ABC (avidin biotinylated horseradish peroxidase complex) peroxidase technique. Fetal cartilage and fracture callus were used as control materials. Type II collagen was present in the matrix of all the cartilage tumors. The reaction was strongest in areas of well-differentiated cartilage and weakest in the poorly differentiated tissue of high-grade chondrosarcomas. Areas of mineralization or ossification, and areas of eosinophilic, fibrous, or degenerated cartilage gave a negative reaction. PMID- 2551431 TI - Early renal artery occlusion after enalapril in atheromatous renal artery stenosis. PMID- 2551432 TI - Treatment of post-herpetic neuralgia. PMID- 2551433 TI - Fructose 2,6-bisphosphate and trehalose metabolism in Saccharomyces cerevisiae. AB - 1. A regulatory mutant of Saccharomyces (fdp) unable to activate fructose 1,6 bisphosphatase presented a normal response to the glucose and fructose signals as measured by trehalase activation, indicating that the inability of the strain to grow on these sugars is caused by a defect located beyond membrane interactions. 2. In vivo experiments with a mutant strain bearing a phosphoglucoisomerase gene (pgil-delta) deletion showed that activation of trehalase and deactivation of the tehalose-6-phosphate synthase complex occurred to the same extent whether glucose or fructose was used as signal. 3. These results suggest that fructose-2,6 bisphosphate is not involved in the interconversion of forms of the enzymes of trehalose metabolism. Furthermore, when fructose-2,6-bisphosphate was assayed on trehalose synthesizing activity using cell-free extracts and partially purified preparations of the complex, no effect was observed. 4. We conclude that regulation by cAMP fulfills the requirements for control of trehalose levels in Saccharomyces. PMID- 2551434 TI - Hemorrhagic cystitis associated with urinary excretion of adenovirus type 11 following allogeneic bone marrow transplantation. AB - We studied a total of 50 recipients who had received allogeneic bone marrow transplantation (BMT) and evaluated both the presence of hemorrhagic cystitis (HC) and the urinary excretion of adenovirus. Twelve recipients developed HC and eight of these 12 patients excreted adenovirus type 11 at the onset of cystitis. Urine for virus isolation was attempted 30, 60 and 100 days after BMT. Among 137 specimens examined, eight were positive for adenovirus type 11. Of these eight samples, six were collected during HC; while in the 129 samples which were negative for adenovirus, only three specimens was collected during HC. Female patients, seropositivity for the antibody to adenovirus prior to BMT and acute graft-versus-host disease (grade 2-4) showed a significant impact on the risk of adenovirus HC. It may be said that adenovirus type 11 is one of the causative agents of HC in BMT recipients. PMID- 2551435 TI - Cytomegalovirus seroconversion in patients receiving intensive induction therapy prior to allogeneic bone marrow transplantation. AB - Cytomegalovirus (CMV) seropositivity in recipients of an allogeneic bone marrow transplant (BMT) is a major risk factor for development of post-transplant CMV infection. CMV serology was assessed in 98 patients during intensive chemotherapy for haematological malignancy prior to allogeneic BMT. Thirty-seven patients eventually received a BMT; the remaining 61 patients were treated with chemotherapy alone. The proportion of seropositive patients in the BMT group increased from 36% to 48% between presentation and transplantation. This represents an increase in recipient seropositivity of 33% as a direct result of pre-transplant therapy. Mean time to seroconversion was 186 days. Seropositivity in patients receiving chemotherapy only increased from 43% to 56% during treatment and follow-up. The most likely source of the CMV acquired by these patients is CMV-infected blood products. We suggest that, wherever possible, CMV negative blood products should be used exclusively from presentation to support all patients receiving chemotherapy in whom BMT is a therapeutic option. PMID- 2551436 TI - Pretransplant herpes virus serology and chronic graft-versus-host disease. AB - The influence of pretransplant herpes virus antibodies in patients and donors in the subsequent development of chronic graft-versus-host disease (GVHD) was analysed in 150 consecutive HLA identical bone marrow recipients. The Cox regression bivariate analysis showed that (i) pretransplant seropositivity for cytomegalovirus (CMV) in the patients and the donors, (ii) donor seropositivity for herpes simplex virus and Epstein-Barr virus, (iii) high donor and patient age, (iv) a previous grade II-IV acute GVHD, (v) patients receiving unirradiated donor buffy coat cells post-transplant, (vi) overall CMV infection, (vii) high donor herpes virus load (positive serology for 3-4 herpes viruses versus 0-2), and (viii) high recipient herpes virus load prior to BMT were all associated with a high incidence of chronic GVHD. In Cox regression multivariate analysis, high pretransplant donor herpes virus load (p less than 0.001) and a previous grade II IV acute GVHD (p = 0.02) were the strongest predictors of chronic GVHD. Thus, herpes virus immune cells in the donated marrow may play a role in the pathophysiology of chronic GVHD. PMID- 2551437 TI - Successful treatment of CMV retinitis with ganciclovir after allogeneic marrow transplantation. AB - Cytomegalovirus (CMV) infection of the retina is a well recognized complication in patients with the acquired immune deficiency syndrome but is rarely seen after bone marrow transplantation (BMT). Among a variety of drugs ganciclovir so far appears to be the most effective therapy for CMV retinitis, but in previous studies relapses occurred in all patients in whom ganciclovir was interrupted. We report the clinical findings in a 22-year-old BMT recipient who developed bilateral exudative CMV retinitis 64 days after BMT despite prophylactic treatment with high-titer CMV-immunoglobulins and transfusions of CMV-negative blood products and donor bone marrow. During a 12 day course of treatment with 7.5 mg/kg/day of ganciclovir the CMV retinitis improved and viruria ceased on day 4 of therapy. In contrast to the previous reports, CMV retinitis in this patient continued to improve even after ganciclovir was stopped and eventually complete healing of all intraretinal lesions as well as total reconstitution of the visual acuity was achieved. He is now free of disease and without relapse of CMV retinitis more than 1 year after transplantation. PMID- 2551438 TI - Gastric damage following local intra-arterial administration of reactive oxygen metabolites in the rat. AB - 1. The effects of reactive oxygen metabolites on the rat gastric mucosa following close-arterial infusion into the left gastric artery have been determined by macroscopic and histological assessment. 2. Local intra-arterial infusion of hydrogen peroxide (0.6-1.3 mumol kg-1 min-1) induced mucosal injury, characterised by areas of pronounced disruption and haemorrhage, which was prevented by concurrent intravenous administration of catalase. 3. Local infusion of the superoxide generating system xanthine-oxidase and hypoxanthine likewise induced extensive haemorrhagic damage and necrosis of the mucosa. Prolonged incubation of this mixture (10 min) before administration, significantly reduced the degree of injury, indicating the lability of the products so formed. 4. The gastric mucosal injury induced by the superoxide generating system was inhibited by concurrent local infusion of superoxide dismutase (96 u kg-1 min-1), as was the associated increase in mucosal permeability to radiolabelled albumin. 5. Administration of catalase did not inhibit the gastric mucosal damage induced by infusion of xanthine oxidase-hypoxanthine, yet augmented the protective effects of a low dose of superoxide dismutase (46 u kg-1 min-1 i.a.). 6. These findings directly confirm that reactive oxygen metabolites can induce extensive gastric mucosal injury, supporting their role in the pathogenesis of gastric damage following ischaemia and hypotensive shock. PMID- 2551439 TI - Non classical, multiple-site interaction of [3H]-prazosin with the alpha 1 adrenoceptor of intact BC3H1 cells. AB - 1. In intact BC3H1 cells the EC50 of noradrenaline (NA) for the inositol phosphate response measured at 37 degrees C (EC50 = 193 nM) was much lower than its apparent dissociation constant (Ki37 degrees C = 83.211 microM) determined at this temperature by [3H]-prazosin binding. 2. After pretreatment of the cells with NA at 37 degrees C for 45 min, the time used in binding assays at this temperature, this difference between EC50 and Ki37 degrees C did not decrease significantly. An agonist-induced reduction in alpha 1-adrenoceptor affinity can therefore not explain the very high Ki37 degrees C value. 3. NA pretreatment at 37 degrees C decreased the number of [3H]-prazosin binding sites (assessed by whole cell binding at 2 degrees C) by only 49%; not by 100%, the value expected if agonist-induced receptor internalization were the origin of the very low Ki37 degrees C. 4. The EC50 of NA for the inositol phosphate response in the presence of 156 pM [3H]-prazosin was 1.841 microM but the IC50 of NA for the inhibition of [3H]-prazosin binding (126 pM) was 316 microM. As there is no alpha 1 adrenoceptor reserve in these cells we propose that at 37 degrees C [3H]-prazosin interacts, not only with the catecholamine recognition site (site 1) of the receptor, but also reacts weakly with another site from which it cannot be directly displaced by catecholamine-like substances (site 2). PMID- 2551440 TI - Superoxide anions enhance platelet adhesion and aggregation. AB - 1. Superoxide dismutase (SOD, 60 u ml-1) or ferricytochrome c (70 microM) significantly inhibited thrombin-stimulated platelet adhesion to gelatin-coated plastic, whereas catalase (1000 u ml-1) or mannitol (1 mM) had no effect. 2. The platelet aggregation induced by low concentrations of thrombin (causing less than 45% maximal change in light transmission) was inhibited by SOD. Catalase or mannitol had no effect on platelet aggregation. 3. Pyrogallol (an O2- generator) enhanced both platelet adhesion to gelatin-coated plastic and platelet aggregation induced by thrombin; this enhancement was neutralized by SOD. 4. These results indicate that O2- increase both platelet adhesion and aggregation, whereas other free radicals such as hydrogen peroxide or hydroxyl radicals are not involved. PMID- 2551442 TI - Analysis of the relaxant effects of AH 21-132 in guinea-pig isolated trachealis. AB - 1. Experiments have been performed with the dual intent of analysing the mechanism by which AH 21-132 relaxes airways smooth muscle and determining whether the effects of this compound can be distinguished from those of theophylline. 2. AH 21-132 (0.25-8 microM) and theophylline (1-1000 microM) each caused concentration-dependent suppression of the spontaneous tone of guinea-pig isolated trachealis. The maximal effect of AH 21-132 was equivalent to that of theophylline. No evidence was obtained that the tissue became sensitized or desensitized to the action of AH 21-132. 3. Propranolol (1 microM) profoundly antagonized the tracheal relaxant action of isoprenaline but not that of AH 21 132. 4. In indomethacin (2.8 microM)-treated tissues, tone was induced by K+-rich (120 mM) Krebs solution, acetylcholine (ACh, 1 mM) or histamine (200 microM). Log concentration-relaxation curves for AH 21-132, isoprenaline and theophylline were all moved to the right in the presence of the spasmogens, the smallest rightward shift being induced by histamine and the greatest by ACh. While maximal effects of AH 21-132 and theophylline were unaffected by the spasmogens, that of isoprenaline was reduced by KCl and ACh. 5. In tissues treated with indomethacin (2.8 microM), AH 21-132 (0.1-100 microM) inhibited the spasmogenic effects of potassium chloride (KCl), ACh and histamine in a concentration-dependent manner. The inhibition was characterized by rightward shifts in the spasmogen concentration-effect curves with depression of their maxima. 6. In tissues treated with both indomethacin (2.8 microM) and ACh (1 mM), the removal of tracheal epithelium caused a small but significant leftward shift in the log concentration-relaxation curve for AH 21-132 but did not alter that for theophylline. 7. In tissues treated with indomethacin (2.8 microM) and maintained at 12 degrees C, theophylline (0.1-3.2 mM) caused concentration-dependent spasm. This effect was not shared by AH 21-132. 8. AH 21-132 (0.1-1000 microM) more potently inhibited the activity of cyclic AMP-dependent than of cyclic GMP dependent phosphodiesterase derived from homogenates of guinea-pig trachealis. Theophylline, too, inhibited these enzymes but was less potent in each case than AH 21-132 and did not exhibit selectivity for the cyclic AMP-dependent enzyme. 9. It is concluded that AH 21-132 exerts a non-specific (i.e. effective no matter what agent is used to support tone) relaxant effect on the trachealis muscle which does not involve the activation of beta l-adrenoceptors. The profile of the relaxant action of AH 21-132 more closely resembles that of theophylline than that of isoprenaline. However, AH 21-132 can be differentiated from theophylline in that: (a) its relaxant potency is increased by epithelial removal; (b) it does not cause tracheal spasm; (c) it exhibits selectivity as an inhibitor of cyclic AMP-dependent as opposed to cyclic GMP-dependent phosphodiesterase. It is possible that the relaxant effects of AH 21-132 are related to its ability to inhibit cyclic nucleotide phosphodiesterases. PMID- 2551441 TI - Angiotensin neuromodulation of adrenergic and purinergic co-transmission in the guinea-pig vas deferens. AB - 1. The effects of angiotensin II (AII) and angiotensin III (AIII) on the isolated vas deferens of the guinea-pig were studied via three parameters: the overflow of adenosine 5'-triphosphate (ATP) and tritiated noradrenaline (NA), the mechanical response to field stimulation and the mechanical response to exogenous NA and alpha, beta-methylene ATP (alpha, beta-mATP). 2. At 2 Hz, AII enhanced the overflow of ATP and NA, whereas at 20 Hz, AII enhanced the overflow of NA but was without significant effect on ATP overflow. AIII, at 2 Hz, inhibited the overflow of ATP, but enhanced NA overflow, whereas at 20 Hz ATP overflow was unaffected, but NA overflow was still enhanced. 3. At 2 Hz, AII enhanced both phases of the response to field stimulation and at 20 Hz the overall response. AIII at 2 Hz enhanced the adrenergic response, but was without effect on the purinergic response to field stimulation. At 20 Hz, AIII was without effect on the overall response. 4. AII enhanced responses to exogenous NA and alpha, beta-mATP, whereas AIII was without effect. 5. These results provide evidence that both ATP and NA release are capable of being modulated by angiotensins. Furthermore, modulation of ATP release is frequency-dependent, whereas [3H]-NA release is not. These results raise questions about the mechanisms of storage and release of the sympathetic co-transmitters NA and ATP; they also show that angiotensin receptors in the guinea-pig vas deferens are not a homogeneous population. PMID- 2551443 TI - Inhibitory effects of AH 21-132 in guinea-pig isolated ileum and taenia caeci. AB - 1. AH 21-132 is being investigated as a potential chemotherapeutic agent for bronchial asthma. The present experiments were designed to determine whether AH 21-132 shares the activity of theophylline as an antagonist at adenosine A1 receptors and to assess its potency as a relaxant in intestinal smooth muscle. 2. In the transmurally-stimulated guinea-pig ileum, theophylline (1 mM), but not AH 21-132 (1 and 10 microM), antagonized twitch depression induced by adenosine. Higher concentrations (100 microM and 1 mM) of AH 21-132 themselves had a depressant effect. Neither theophylline (1 mM) nor AH 21-132 (1 and 10 microM) antagonized twitch depression induced by noradrenaline. 3. AH 21-132 (100 microM and 1 mM) depressed maximum contractions of ileum induced by both acetylcholine (ACh) and histamine. 4. In ileum treated with hyoscine (1 microM), AH 21-132 (greater than 10 microM) caused a concentration-dependent depression of the log concentration-effect curve for potassium chloride. 5. Simultaneous extracellular electrophysiological and mechanical recording from taenia caeci showed that AH 21 132 (100 microM-1 mM) inhibited spontaneous tension waves and their associated bursts of electrical spike activity. 6. Intracellular electrophysiological recording from taenia caeci showed that the mechano-inhibitory effect of 1 mM AH 21-132 was accompanied by abolition of spontaneous spike activity. Following spike abolition, the membrane potential assumed a value very close to that observed during periods of electrical quiescence prior to drug exposure. 7. AH 21 132 inhibited the activity of cyclic AMP-dependent and cyclic GMP-dependent phosphodiesterases derived from homogenates of ileal smooth muscle. The effective concentration ranges were 0.1-1OOO microM and 1-1000 microM, respectively. Theophylline, too, inhibited these enzymes but in each case was less potent than AH 21-132. 8. It is concluded that AH 21-132 is devoid of antagonist activity at adenosine Al receptors which modulate ACh release from intramural cholinergic nerves in the ileum. At concentrations greater than IO microM, AH 21-132 has a relaxant effect on intestinal smooth muscle characterized by suppression of spontaneous action potentials but by minor change in resting membrane potential. AH 21-132 previously has been reported to depress the spontaneous tone of trachealis muscle with an EC50 value of less than lO microM and the present experiments therefore show that this agent is much less potent in inhibiting intestinal muscle. This potency difference cannot be attributed to a tissuerelated difference in the potency of AH 21-132 as an inhibitor of cyclic AMP- or cyclic GMPdependent phosphodiesterases. PMID- 2551444 TI - Inhibition of platelet-activating factor- and zymosan-activated serum-induced chemotaxis of human neutrophils by nedocromil sodium, BN 52021 and sodium cromoglycate. AB - 1. Inflammatory cells such as eosinophils and neutrophils are thought to contribute actively to the pathogenesis of asthma since they infiltrate into the lung tissue. These cells are mobilized by lipid-like and protein-like chemotactic factors. As illustrative examples of both groups, platelet-activating-factor (Paf) and zymosan-activated-serum (ZAS) were used in this study. The inhibitory effects of nedocromil sodium, the Paf antagonist BN 52021 and sodium cromoglycate on Paf- and ZAS-induced neutrophil chemotaxis were evaluated. 2. All tested drugs inhibited Paf-induced neutrophil chemotaxis with approximately the same potency (IC50 approximately 1 nM). 3. Nedocromil sodium and sodium cromoglycate were equally potent in inhibiting ZAS-induced neutrophil chemotaxis (IC50 = 0.1-1 microM), whereas BN 52021 was considerably less potent (IC30 = 10 microM). 4. To find out whether the drugs tested could inhibit early events in cell activation, their capacity to inhibit Paf- and ZAS-induced cytosolic free Ca2+-mobilization was investigated. BN 52021, at a concentration of 100 microM, completely inhibited Paf-induced Ca2+-mobilization and inhibited ZAS-induced Ca2+ mobilization by about 50%. Nedocromil sodium and sodium cromoglycate were ineffective. PMID- 2551445 TI - 3-Aminopropylphosphinic acid--a potent, selective GABAB receptor agonist in the guinea-pig ileum and rat anococcygeus muscle. AB - 1. 3-Aminopropylphosphinic acid, a gamma-aminobutyric acid (GABA) analogue, was tested for activity on guinea-pig isolated ileum and rat isolated anococcygeus muscle preparations. The effects of 3-aminopropylphosphinic acid were compared with those of GABA and baclofen. 2. In the electrically stimulated ileum, 3 aminopropylphosphinic acid, like GABA and baclofen, caused a concentration dependent inhibition of the cholinergic twitch contraction, the IC50 value being 1.84 +/- 0.23 microM (n = 12). Unlike GABA, but like baclofen, 3 aminopropylphosphinic acid did not produce an initial contraction. 3. The inhibitory effects of 3-aminopropylphosphinic acid and baclofen in the guinea-pig ileum were not significantly antagonized by bicuculline (10 microM), phentolamine plus propranolol (both 1 microM), yohimbine (1 microM), naloxone (1 microM), impromidine (1 microM) or 8-phenyltheophylline (10 microM). The inhibitory effects of 3-aminopropylphosphinic acid, but not of baclofen, were however antagonized by phaclofen (500 microM). In addition the effects of 3 aminopropylphosphinic acid were abolished by baclofen desensitization in the guinea-pig ileum. 4. 3-Aminopropylphosphinic acid, GABA and baclofen reduced the twitch contraction evoked by electrical field stimulation in the rat anococcygeus muscle. The IC50 for 3-aminopropylphosphinic acid inhibition of the anococcygeus contraction was 0.89 +/- 0.15 microM (n = 8). 5. It is concluded that 3 aminopropylphosphinic acid is a potent, selective GABAB agonist, being seven times more potent than baclofen in the guinea-pig ileum and five times more potent than baclofen in the rat anococcygues muscle preparations. PMID- 2551447 TI - Perinatal lead exposure impairs opioid but not non-opioid stress-induced antinociception in developing rats. AB - 1. The development of opioid systems has been shown to be sensitive to perinatal exposure to lead. We have studied the effects of such exposure on opioid and non opioid mediated stress-induced antinociception in developing rats. 2. Lead was administered in the maternal drinking water from conception to postnatal day 14 at 300 and 1000 p.p.m. Twenty and 25 day old rats were subjected to swimming stress and antinociception measured using the tail immersion test. 3. A 3 min swim-stress induced an opioid-mediated antinociceptive response in 20 day old rats which was attenuated by 300 p.p.m. lead and by 1000 p.p.m. lead treatment in a dose-related manner. A 10 min swim-stress induced a non-opioid mediated antinociceptive response in 25 day old rats which was not antagonised by 300 or 1000 p.p.m. lead. 4. Naloxone antagonised the residual antinociception observed in 20 day old animals treated with 300 p.p.m. lead and had no effect on antinociception in control or lead-treated 25 day old rats. 5. Using a lead exposure model considered to represent subclinical lead toxicity in man, it was shown that perinatal lead exposure disrupts opioid but not non-opioid mediated stress antinociception. PMID- 2551446 TI - The pharmacological properties of the peptide, endothelin. AB - 1. The effect of endothelin (ET-1) has been studied on isolated vascular and non vascular preparations, using both functional and competition radioligand binding techniques. The effects of endothelin on blood pressure were studied in both anaesthetised, chemically denervated normotensive and spontaneously hypertensive rats (SHR). 2. Endothelin elicited contractile responses in the rat thoracic aorta, perfused mesenteric bed, rabbit mesenteric artery and portal vein. The maximal responses in the rat aorta were enhanced by removal of the endothelium, and were reduced in the presence of either a cyclo-oxygenase inhibitor (indomethacin) or a thromboxane receptor antagonist (SQ 29,548). In terms of potency, the most sensitive preparation was the rat endothelium-denuded aorta and rat perfused mesenteric bed (-log EC50 values = 8.2 +/- 0.07 and 8.2 +/- 0.12, mean +/- s.e.mean, n = 4, respectively). In the perfused mesenteric bed of the rat the maximum response to endothelin (219 +/- 12 mmHg, n = 4) was greater than that to either phenylephrine (maximal response = 67 +/- 9 mmHg; n = 4) or KCl (maximal response = 110 +/- 6 mmHg, n = 4). 3. Endothelin elicited contractile responses of the guinea-pig isolated ileum, oesophageal muscularis mucosae and uterus. Responses were also observed in the rat fundic strip and paced left atria. The guinea-pig urinary bladder, trachea, rat vas deferens and anococcygeus exhibited little or no response to endothelin at the concentrations studied (1 x 10(-12)-3.2 x 10(-8) M). Of the above preparations, the ileum and oesophageal muscularis mucosae were the most sensitive to endothelin (-log EC50 = 8.5 +/- 0.11 and 8.4 +/- 0.06, n = 6, respectively), exhibiting potencies similar to those observed in the endothelium-denuded aorta of the rat. 4. In competition radioligand binding studies, endothelin did not displace either [3H]-PN 210-100 or [125I]-(-)-omega-conotoxin GVIA from binding sites in membranes from rat cerebral cortex and, skeletal muscle or from guinea-pig cerebral cortex and hippocampus, respectively. This indicates a lack of direct interaction of endothelin at the dihydropyridine binding site and the N-type calcium channel, respectively. However, in functional studies, contractile responses to endothelin (1 x 10(-8) M) in the endothelium-denuded aorta of the rat were potently reversed by nifedipine, verapamil, and prenylamine (-log IC50 values = 8.0 +/- 0.13, 7.2 +/- 0.09 and 6.6 +/- 0.08, n = 4-8, respectively). In addition, the responses to endothelin were virtually abolished in the presence of Krebs physiological salt solution containing no calcium but with 1 x 10-M EDTA added. Preequilibration with either (-)-w-conotoxin (1 x 10-6M) or tetrodotoxin (1 x 10-6M) did not affect responses to endothelin. 5. In chemically denervated rats, endothelin (1pmolkg-'-10nmolkg- , i.v.) exhibited pressor responses, which were unaffected by a 3 h pretreatment with indomethacin. In the SHR, the effects on blood pressure were not significantly different from those observed in normotensive animals at any of the doses studied. A transient (duration < 30 s) depressor response was also observed in all groups studied at a dose of 0.1-1 nmol kg-1 i.v. 6. In conclusion, endothelin is a potent contractile agonist in both vascular and non-vascular muscle. It appears to elicit responses partly via the entry of extracellular calcium (by a mechanism distinct from that of other calcium facilitators) and partly by release of endoperoxides. PMID- 2551449 TI - Back pain in women with chronic pelvic inflammatory disease and inflammatory sacroiliac disease. PMID- 2551448 TI - Comparison of the sodium currents in normal Purkinje fibres and Purkinje fibres surviving infarction--a pharmacological study. AB - 1. Purkinje fibres surviving infarction showed a lower maximum upstroke velocity (Vmax) and a longer action potential duration when compared to normal Purkinje fibers. A reduction in the fast sodium current and an increase in the sodium 'window' current may be responsible for the observed alterations in Vmax and action potential duration respectively. 2. Since voltage clamp studies were not feasible, a pharmacological approach was used. The responses to tetrodotoxin (TTX) and lignocaine in normal Purkinje fibres and Purkinje fibres surviving infarction were used to examine the sodium currents in these fibres. 3. Vmax, an indirect measure of the fast sodium current, was more sensitive to lignocaine in Purkinje fibres surviving infarction than in normal Purkinje fibres. The reduction in Vmax by lignocaine was more prominent at the shorter stimulation cycle length. Significant reduction of Vmax was observed with the higher concentration of TTX and no differential effect on Vmax between normal Purkinje fibres and Purkinje fibres surviving infarction was detected. 4. Reduction of action potential duration in the presence of TTX or lignocaine was used as a measure of the sodium 'window' current. A greater reduction of action potential duration by TTX and lignocaine was observed in normal Purkinje fibres than in Purkinje fibres surviving infarction. 5. The results suggested that the fast sodium current in Purkinje fibres surviving infarction is more sensitive to pharmacological agents with local anaesthetic properties and the prolonged action potential duration in these Purkinje fibres cannot be due to an increase in the sodium 'window' current. The results are compatible with an enhanced effect of antiarrhythmic drugs on Vmax and conduction in the ischaemic heart. PMID- 2551450 TI - ATP-sensitive chemoreceptors: antagonism by other nucleotides and the potential implications of ectonucleotidase activity. AB - As measured by extracellular single-cell recording, the responses to adenosine triphosphate (ATP) by ATP-sensitive chemoreceptors (ATP cells) on the olfactory organ of the spiny lobster are markedly suppressed by adenosine diphosphate (ADP), adenosine monophosphate (AMP) and to a lesser extent, adenosine, when each is presented in binary mixture with ATP. In the presence of ADP, the dose response function for ATP exhibits an apparent parallel displacement to the right suggesting that this antagonism may occur via competition at the ATP receptor. Structure-activity relationships reveal that the structural requirements for antagonism by diphosphate analogs of ADP bear little relationship to the requirements for the agonistic activity of corresponding triphosphate analogs. Under Mg2+-free conditions, the desensitization of ATP cells tends to be delayed resulting in enhanced responses to ATP. Desensitization does not appear to be related to the generation of the antagonist, ADP, from ATP via ecto-ATPase activity. The results of this study suggest that the responses of ATP cells to the ATP contained in natural stimulus (odor) mixtures can be tempered by the suppressive interactions of other nucleotides in the mixtures. Furthermore, these interactions may be mitigated and/or intensified by the actions of sensillar ectonucleotidases. PMID- 2551451 TI - Schild plot analysis of glycine and kynurenic acid at the N-methyl-D-aspartate excitatory amino acid receptor. AB - Glycine enhanced the N-methyl-D-aspartate (NMDA)-stimulated sodium flux in rat hippocampal slides in a concentration-dependent manner. The potentiation by glycine was apparently not competitive to NMDA with a maximal effect of about 50% enhancement at approximately 150 microM glycine. Glycine also reversed the kynurenic acid inhibition of NMDA-stimulated sodium flux, increasing flux two fold. In the absence of glycine, the kynurenic acid inhibition of NMDA-stimulated sodium flux appeared not competitive to NMDA, with a Schild plot slope of 0.6 +/- 0.1, significantly less than 1 (P less than 0.05). Addition of 100 microM glycine gave a Schild plot slope of 1.0 +/- 0.3, classically defining competitive inhibition. However, the pKb of 4.5 obtained from the x intercept gives an inaccurate estimate of the affinity of kynurenic acid for the NMDA recognition site since glycine is present. Addition of 400 microM glycine resulted in a Schild plot slope of 1.9 +/- 0.2. By varying the concentrations of glycine, we have apparently reproduced a portion of the theoretical set of curves for various ratios of affinities of the antagonist for the agonist recognition site and a second site. These results support a model where glycine modulates NMDA receptor function and where kynurenic acid acts both as a competitive antagonist to NMDA and at a second site from which it is displaced by glycine. PMID- 2551452 TI - Neurotoxicity of beta-N-methylamino-L-alanine (BMAA) and beta-N-oxalylamino-L alanine (BOAA) on cultured cortical neurons. AB - Recent studies have implicated the ingestion of the structurally related plant excitotoxins, beta-N-methylamino-L-alanine (BMAA), and beta-N-oxalylamino-L alanine (BOAA), in the pathogenesis of two human motor system diseases, the amyotrophic lateral sclerosis-Parkinsonism-dementia complex of Guam (Guam ALS PD), and lathyrism, respectively. We have investigated the toxicity of these amino acids on cultured mouse cortical neurons in the presence of physiological concentrations of bicarbonate (a required toxic cofactor for BMAA neurotoxicity). A 24 h exposure to 10 microM - 3 mM BMAA, or to 300 nM - 100 microM BOAA, induced, concentration-dependent neuronal degeneration without glial damage; the neurotoxic EC50 for BMAA was about 1 mM, and the EC50 for BOAA was about 20 microM. At high concentrations, both compounds destroyed essentially the entire neuronal population. Neurotoxicity also depended on exposure duration, with reduced injury at an exposure time of 1 h, and increased injury at an exposure time of 3 days. Despite the fact that ingestion of BMAA and BOAA both lead to motor system damage, previous studies have suggested that the two excitotoxins act primarily on different glutamate receptor subtypes: BMAA on N-methyl-D aspartate (NMDA) receptors, and BOAA on non-NMDA receptors. Consistent with these studies, the neurotoxicity of high concentrations of BMAA was substantially attenuated by 1 mM D-amino-5-phosphonovalerate (D-APV), whereas BOAA neurotoxicity was less sensitive to D-APV but was attenuated by 2 mM kynurenate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551453 TI - gamma-Aminobutyric acid inhibits the release of alpha-melanocyte-stimulating hormone from rat hypothalamic slices. AB - The effect of gamma-aminobutyric acid (GABA) on release of alpha-melanocyte stimulating hormone (alpha-MSH) from hypothalamic neurons was investigated in vitro using the perifusion technique. Rat hypothalamic slices were continuously superfused with Krebs-Ringer medium and the release of alpha-MSH in the effluent perifusate was monitored by means of a sensitive and specific radioimmunoassay method. Infusion of 50 mM K+ for 15 min induced a transient increase of alpha-MSH release (5- to 8-fold above the spontaneous level). Infusion of the same dose of K+ for 75 min caused a brief discharge of alpha-MSH during the first 30 min followed by sustained release of the neuropeptide. The effect of GABA was investigated 27 min after the onset of KCl infusion. Application of GABA (5 x 10( 5) M) resulted in a significant and reversible inhibition of K+-induced alpha-MSH release. The GABAA agonist, muscimol (10(-4) M), produced a prolonged inhibition of K+-evoked alpha-MSH release, while the GABAB agonist, baclofen (10(-4) M), was devoid of effect on hypothalamic alpha-MSH release. Bicuculline (10(-4) M), a specific GABAA antagonist, had no effect when added alone to the medium but totally reversed the inhibitory effect of GABA on K+-induced alpha-MSH release. Taken together, these data suggest that exogenous GABA exerts an inhibitory control on alpha-MSH neurons. Our data also show that the effect of GABA on alpha MSH release by hypothalamic neurons is mediated through GABAA-type receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551454 TI - Participation of opiate receptors located in the nucleus tractus solitarii in the hypotension induced by alpha-methyldopa. AB - The local administration of the opiate receptor antagonist, naltrexone, into the nucleus tractus solitarii (NTS) inhibited the hypotension induced by systemically injected alpha-methyldopa in conscious rats. In addition, the local injection into the NTS of a beta-endorphin antiserum but not of antisera against [Met5]enkephalin and dynorphin A(1-13) prevented the alpha-methyldopa-induced hypotension. These results suggest a role of opiate receptors in the NTS, or in a closely located medullary site, in the centrally mediated hypotension induced by alpha-methyldopa. PMID- 2551455 TI - Alpha-chloralose opens the chloride channel of frog isolated sensory neurons. AB - The effect of alpha-chloralose on the sensory neurons isolated enzymatically and mechanically from frog dorsal root ganglia was studied using a suction-pipette technique. The threshold concentration of alpha-chloralose was around 3 x 10(-5) M and the current produced by alpha-chloralose saturated at the concentration of 3 x 10(-3) M or more. The dose-response curve for alpha-chloralose provided a Ka value of 6 x 10(-4) M and a Hill coefficient of 1.8. The reversal potential of the response elicited by alpha-chloralose was close to the equilibrium potential for Cl- (ECl), indicating that the current was carried through Cl- channels. The current-voltage relationship indicated that there was little voltage dependence in the alpha-chloralose-induced response. The analysis of the variance of the alpha-chloralose-induced Cl- current fluctuations showed two types of the receptor-ionophore complexes with different channel conductances. PMID- 2551456 TI - The effect of the excitatory amino acid receptor antagonist dizocilipine maleate (MK-801) on hemispheric cerebral blood flow and metabolism in dogs: modification by prior complete cerebral ischemia. AB - The effect of the N-methyl-D-aspartate (NMDA) receptor antagonist dizociplipine maleate (MK-801) on cerebral blood flow (CBF), cerebral metabolic rate for oxygen (CMRO2), intracranial pressure and systemic variables was examined in 6 normal dogs (Group I). In 6 additional dogs (Group II), the effects of a prior 11 min episode of complete cerebral ischemia on the response to dizocilipine was studied. CBF was measured with a sagittal sinus outflow technique and CMRO2 was calculated as the product of CBF and the arterial to sagittal sinus O2 content difference. Dizocilipine was administered as a 150 micrograms/kg i.v. bolus followed by a 75 micrograms.kg-1.h-1 infusion for 90 min. Plasma dizocilipine levels were greater than 25 ng/ml for the duration of the infusion. The CSF levels were approximately half the plasma levels. Five minutes after initiation of dizocilipine treatment, Group I dogs experienced a 63% increase in heart rate (P less than 0.01) and an 8% decrease in the mean arterial blood pressure (P less than 0.05). Over the same time interval. CBF increased by 85% (P less than 0.01) and intracranial pressure nearly doubled (P less than 0.05). In addition, dizocilipine treatment in all Group I animals resulted in EEG quasiperiodic bursts of delta-waves and polyspikes on a background of beta-activity. With the exception of the intracranial pressure, the above changes in systemic and cerebral variables persisted for the duration of the drug infusion. Intracranial pressure was no longer significantly elevated after 80 min of drug infusion. Hemispheric CMRO2 was unchanged by dizocilipine in Group I dogs. There was a decrease in the cortical glucose level at the end of the study, but no significant change in phosphocreatine, ATP, lactate, or energy charge when compared with 6 laboratory normals. An identical dose of dizocilipine administered after an 11 min episode of complete cerebral ischemia resulted in no significant changes in either cerebral or systemic variables. The absence of systemic effects in Group II dogs suggests that dizocilipine administration in normal dogs results in a centrally mediated activation of the peripheral sympathetic nervous system. The uncoupling of CBF and CMRO2 observed following dizocilipine treatment is similar to that reported for two other known NMDA antagonists, ketamine and phencyclidine. If administration of dizocilipine results in improved histopathological and neurological outcome following an episode of complete cerebral ischemia, this improvement is unrelated to changes in postischemic CBF or hemispheric CMRO2. PMID- 2551457 TI - Suppressive effects of intranigral injection of muscimol in three models of generalized non-convulsive epilepsy induced by chemical agents. AB - The involvement of intranigral gamma-aminobutyric acid (GABA) receptors in the control of generalized non-convulsive epilepsy was investigated in the rat in 3 models of petit mal epilepsy induced by systemic administration of gamma butyrolactone, pentylenetetrazol and 4,5,6,7-tetrahydroisoxazolo [5,4-c]pyridin 3 ol (THIP). Bilateral intranigral injection of muscimol (2 ng/0.2 microliters/side), a GABAA receptor agonist, significantly reduced the duration of EEG-recorded spike-and-wave discharges induced by gamma-butyrolactone (100 and 200 mg/kg i.p.), pentylenetetrazol (20 mg/kg i.p.) and THIP (7.5 mg/kg i.p.). This treatment had no effect on the electroencephalographic discharges observed after injection of THIP (10 mg/kg i.p.). Bilateral injection of muscimol (2 and 4 ng/side) into the substantia nigra did not modify the latency of onset nor the duration of clonic seizures induced by pentylenetetrazol at the dose of 40 mg/kg i.p. Bipolar depth electrode recording indicated that intranigral injection of muscimol did not alter nigral electroencephalographic activity. Autoradiography following intranigral injection of [3H]muscimol indicated a diffusion not exceeding 400 microns from the injection site. These results confirm that activation of GABA receptors in the substantia nigra suppresses the occurrence of spike-and-wave discharges in animal models of generalized non-convulsive epilepsy. PMID- 2551458 TI - Epileptiform activity in vitro can produce long-term synaptic failure and persistent neuronal depolarization. AB - A large, extracellular negative DC shift, termed epileptic depolarization, could be elicited during zero magnesium-induced epileptic activity in the rat hippocampal slice. In 10 mM glucose medium, epileptic depolarization was elicited by high-frequency synaptic stimulation. During epileptic depolarization synaptic responses were abolished, but recovered in 10.4 +/- 2.1 min. In low glucose (2 mM) medium, epileptic depolarization either occurred spontaneously or could be elicited by high frequency synaptic stimulation, and no recovery of synaptic responses was observed for at least 30 min. This long-term synaptic failure was blocked by the competitive NMDA antagonists, 3-[+/-)-2-carboxypiperazin-4-yl) propyl-1-phosphonate (CPP, 100 microM) and D-2-amino-7-phosphonoheptanoate (D AP7, 100 microM) when added at the peak of epileptic depolarization, but not 5 min afterwards. Intracellular analysis showed that this extracellular DC shift was correlated with a membrane depolarization which approached 0 mV. With 10 mM glucose medium, the membrane potential returned to resting level in 6.3 +/- 1.9 min. In 2 mM glucose medium, neurons remained depolarized and no recovery was observed. This persistent depolarization could account for the loss of synaptic function recorded extracellularly. Application of 100 microM CPP blocked persistent depolarization and allowed for the recovery of the membrane potential. Epileptic depolarization was also observed during picrotoxin-induced epileptic activity. Both anoxic depolarization during experimental ischemia and epileptic depolarization can trigger long-term synaptic failure and persistent depolarization. Epileptic depolarization and anoxic depolarization may be triggers which can lead to neuronal failure in diseases associated with neuronal degeneration. PMID- 2551459 TI - Effects of low- and high-intensity exercise on plasma and cerebrospinal fluid levels of ir-beta-endorphin, ACTH, cortisol, norepinephrine and glucose in the conscious dog. AB - This study was designed to assess effects of exercise on plasma and cerebrospinal fluid (CSF) levels of immunoreactive (ir) beta-endorphin, ACTH, cortisol, norepinephrine, and glucose in the conscious dog. Dogs were exercised on a treadmill at low or high intensity (4.2 miles/h and a 6% or 20% incline) for 90 min, and were allowed to recover for 90 additional min. Neither intensity of exercise changed plasma glucose levels, but dose-related changes in glucose kinetics did occur. CSF glucose declined in both groups. During low intensity exercise, plasma levels of ir-beta-endorphin, ACTH, and cortisol increased with duration of exercise. During high intensity exercise, ACTH, ir-beta-endorphin and cortisol increased faster, and the integrated plasma response of these hormones was greater. Thus, peripheral release of ir-beta-endorphin, ACTH, and cortisol during exercise is dose-related with respect to time and intensity. CSF ir-beta endorphin and ACTH both increased during low- but not high-intensity exercise. CSF cortisol rose markedly in both exercise groups. During high-intensity exercise there was a 50% increase in CSF norepinephrine, indicating that exercise induces alterations in central noradrenergic turnover. We conclude that exercise is a physiologic regulator of both peripheral and central neuroendocrine systems. PMID- 2551460 TI - Time- and frequency-dependent effects of potassium channel blockers on large and medium diameter optic tract axons. AB - Compound action potential recording techniques were used to investigate the time- and frequency-dependent effects of 4-aminopyridine (4-AP) and tetraethylammonium (TEA) on large diameter, fast conducting (t1) and medium diameter, middle conducting (t2) optic tract axons in anesthetized hooded rats. Single-pulse studies show 4-AP causes a rapid decrease in t1 and t2 response amplitude with larger decreases and longer lasting effects in t2 axons. In both axons, 4-AP leads to waveform broadening which is accounted for by increases in fall time since rise time and conduction velocity are unaffected by 4-AP. Strength-duration curves reveal 4-AP increases rheobase and decreases chronaxie in both axons with larger increases occurring in t1. T1, but not t2, axons also display some TEA sensitivity. The absolute and relative refractory periods, determined with paired pulse recovery functions, are increased by 4-AP to a greater degree in t1 than t2 axons. These axons, however, display equal sensitivity to TEA. In contrast, 4-AP and TEA decrease frequency following in both axons with larger effects observed in t2. Based on our data, and that of others, we speculate that t1 axons exhibit 4-AP and TEA sensitivity at nodal/paranodal regions and not at internodal regions, while t2 axons exhibit 4-AP sensitivity at nodal/paranodal and internodal regions and TEA sensitivity only at internodal regions of the axolemma. The possible relevance of these findings to the distribution of 4-AP- and TEA-sensitive potassium channels on t1 and t2 axons and the coding of visual spatial and temporal information remains to be determined. PMID- 2551461 TI - Neuropeptide Y-induced effects on hypothalamic corticotropin-releasing factor content and release are dependent on noradrenergic/adrenergic neurotransmission. AB - Neuropeptide Y (NPY) administration increases both hypothalamic corticotropin releasing factor-like immunoreactivity (CRF-ir) and plasma adrenocorticotropin (ACTH). The dependence of these effects on noradrenaline and adrenaline was investigated by selectively depleting these neurotransmitters with 6 hydroxydopamine (6-OHDA) prior to administration of NPY. This combined treatment decreased hypothalamic CRF-ir (P less than 0.025), an effect isolated to the median eminence (P less than 0.025), whereas plasma ACTH increased greatly compared to 6-OHDA treatment alone (P less than 0.0005). In order to further investigate the potential mechanism of this NPY effect, the alpha 2-adrenergic agonist clonidine was administered to normal rats. This treatment increased plasma ACTH (P less than 0.005) and decreased hypothalamic CRF-ir (P less than 0.025), an effect localized to the median eminence (P less than 0.01). The results from both of these treatments are consistent with increased release of hypothalamic CRF. These data imply that the NPY-induced effects are dependent on normal noradrenergic/adrenergic neurotransmission. Depletion of these neurotransmitters allowed NPY to profoundly stimulate CRF release with no evidence for alteration in synthesis, a result common to alpha 2 stimulation. PMID- 2551462 TI - Differential effects of chronic treatment with mianserin and protryptiline on rat brain cortical alpha 1-adrenoceptors. AB - Chronic administration of mianserin induced an increase in the density of [3H] prazosin binding sites (23%) in membranes and in the maximal noradrenaline stimulation of phosphoinositide breakdown (81%) in slices from rat brain cortex. In contrast, a similar treatment with protryptiline did not induce any significant changes. These findings suggest that the effects of antidepressant drugs on rat brain cortical alpha 1-adrenoceptors may depend on the characteristics of the drug used. PMID- 2551463 TI - Transient forebrain ischemia produces multiple deficits in dopamine D1 transmission in the lateral neostriatum of the rat. AB - Striatal dopamine D1 transmission was studied in rats 7 days after transient (30 min) forebrain ischemia using the 4-vessel occlusion model. The striatal distribution of dopamine D1 ([3H]SCH 23390 binding sites) and D2 ([3H]sulpiride binding sites) receptors as well as the distribution of adenylate cyclase ( [3H]forskolin binding sites) and of the intracytoplasmic dopamine and cAMP regulated phosphoprotein DARPP-32 related to D1 transmission were analyzed. While the distribution of D2 receptors was unaffected 7 days after the ischemic insult, all the other markers showed a patchy disappearance in the dorsolateral part of the neostriatum. These findings underline the existence of selective multiple deficits in D1 transmission after transient forebrain ischemia in rat striatum. PMID- 2551465 TI - Projections from visual cortical areas of the superior temporal sulcus to the lateral terminal nucleus of the accessory optic system in macaque monkeys. AB - Tritiated amino acids were injected into the striate area and in single visual areas of the superior temporal sulcus (STS) of 7 cynomolgus monkeys, in order to trace visual cortical projections to the nuclei of the accessory optic system (AOS). Injections in STS separately involved the areas MT and MST, and resulted in labels within the lateral terminal nucleus of the AOS. In no case were labels found within the AOS nuclei in the brains injected in the striate area, or within the contralateral AOS. It seems likely that the areas MT and MST contribute signals--selectively related to visual motion processing--to the AOS, which is probably involved in the neuronal pathway subserving the optokinetic reflex. PMID- 2551464 TI - Facilitation of ACTH secretion by morphine is mediated by activation of CRF releasing neurons and sympathetic neuronal pathways. AB - Exogenously applied opioid agonists have a stimulatory effect on adrenocorticotropic hormone (ACTH) secretion. The present experiments were designed to examine the mechanisms involved in the stimulatory effect of the mu receptor agonist morphine on ACTH release in chronically cannulated, freely moving, non-stressed rats. Morphine (7.5 mg/kg, i.v.) treatment was followed by a significant increase in plasma levels of ACTH. Pretreatment with the peripheral ganglionic blocker chlorisondamine (3 mg/kg, i.p.) attenuated the response to morphine. The morphine stimulatory effect was also partially inhibited if the rats were pretreated with a specific antiserum to corticotropin-releasing factor (CRF). In rats given both CRF antiserum and chlorisondamine, the plasma ACTH levels remained unchanged after morphine application. These findings indicate that morphine stimulates the release of ACTH by activating both CRF-secretion and peripheral sympathetic neuronal pathways. PMID- 2551466 TI - Loss of NGF receptor immunoreactivity in basal forebrain neurons of aged rats: correlation with spatial memory impairment. AB - Nerve growth factor (NGF) has recently been implicated as a trophic agent in the survival and maintenance of basal forebrain cholinergic neurons. To test the hypothesis that NGF may play a role in the age-related decline of cerebral cholinergic function and loss of cognitive ability, we investigated the possible correlation between the loss of basal forebrain neurons that stain for NGF receptor, and impairment of spatial reference memory performance in aged rats. Our results suggest that NGF receptor-positive basal forebrain neurons undergo marked cell atrophy and loss of neuropil staining in aged rats exhibiting impaired spatial learning and memory performance. Conversely, numerous, densely immunoreactive perikarya and a profuse neuritic plexus within the basal forebrain nuclei was consistently observed in behaviorally intact rats. Overall, the mean number of NGF receptor-positive basal forebrain neurons both in the nucleus of the diagonal band and nucleus basalis correlated with retention of the spatial task (r = 0.84 and r = 0.67, respectively; P less than 0.01). Our results support the view that progressive failure of retrograde trophic support due to the age related loss of NGF receptors may promote degenerative changes in basal forebrain cholinergic neurons, and contribute to deterioration of cognitive ability in senescence. PMID- 2551467 TI - Further studies on the identification of microglia in mixed brain cell cultures. AB - The investigation of brain microglia in primary cultures of dissociated cerebral cortical cells has been continued here utilizing a histochemical procedure for thiamine pyrophosphatase, which selectively stains microglia in whole tissue. This procedure also selectively stained a subpopulation of cells in the cortical cultures. The stained cells were small and exhibited quite variable morphology; from these features, a corresponding cell population could be identified in viable cultures. The stained cells, which were distinct from previously identified macrophage forms, were also distinguished from similar appearing cell types--small neurons and oligodendrocytes. Based on their staining properties, morphology, and distinction from other cell types, the identified cells are proposed as equivalents of ramified microglia. PMID- 2551468 TI - Site-selective cyclic AMP analogs as new biological tools in growth control, differentiation, and proto-oncogene regulation. AB - The physiologic role of cyclic adenosine monophosphate (cAMP) in the growth control of a spectrum of human cancer lines, including leukemic lines, and v-rasH oncogene-transformed NIH/3T3 cells is demonstrated by the use of site-selective cAMP analogs. These cAMP analogs, which can select either of the two known cAMP binding sites of the cAMP receptor protein, induce potent growth inhibition, phenotypic change, and differentiation (leukemic cells) of cancer cells at micromolar concentrations with no sign of cytotoxicity. The growth inhibition parallels selective modulation of cAMP-dependent protein kinase isozymes, type I versus type II, and suppression of cellular proto-oncogene expression. Site selective cAMP analogs thus provide new biological tools for investigating cell proliferation and differentiation and also for the improved management of human cancers. PMID- 2551469 TI - Effects of adding nedocromil sodium (Tilade) to the routine therapy of patients with bronchial asthma. AB - In a 12-week double-blind, group comparative trial, preceded by a 2-week baseline period, 38 asthmatic subjects of mixed aetiology and varying severity received either 4 mg nedocromil sodium by metered dose inhaler twice a day or a matching placebo preparation, in addition to their existing maintenance therapy of inhaled corticosteroids plus inhaled bronchodilators. Asthma severity and lung function were assessed at 4-weekly clinic visits, and symptomatology (morning tightness, daytime asthma, cough, night-time asthma), morning, afternoon and evening PEFR, and the use of inhaled bronchodilators were recorded on daily diary cards. Treatment with nedocromil sodium led to significant (P less than 0.05) improvements in clinic assessment of FEV1 and PEFR both before and after an inhaled bronchodilator from at least the eighth week onwards. Mid-study FVC was also significantly (P less than 0.05) improved. Daily PEFR increased throughout the study in the nedocromil sodium-treated subjects and the diurnal variation was reduced. Daily symptom severity was also reduced and these improvements occurred despite the similar or slightly reduced use of inhaled bronchodilators. However, none of these improvements in diary card parameters reached statistical significance. By the final week of the study subjects treated with nedocromil sodium predominantly had a mild form of asthma or no symptoms at all, and both patients and clinicians reported the effectiveness of nedocromil sodium; the subjects but not the clinicians finding it significantly more effective (P less than 0.05) than placebo. Nedocromil sodium was well tolerated although one patient was withdrawn owing to a persistent sore throat after 7 weeks of treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551470 TI - Nedocromil sodium in bronchial antigen challenge. PMID- 2551471 TI - [Small cell lung cancer: survival analysis of 96 cases]. AB - Existing records of 96 patients with small cell lung cancer (SCLC) at TSGH from 1983 to 1987 were analyzed to determine the relationship of survival rate and prognostic factors including age, sex stage, symptomatic history, liver metastasis, bone metastasis, albumin level, hemoglobin value, and choice of treatment. For the SCLC patients the median survival time (MST) is only 7.2 month, the 1-year survival rate is 25.1%, and the 2-year survival rate is 5.1%. The prognosis for patients with liver or bone metastasis was poor. Sixteen prognostic variables were evaluated in a Cox proportional hazard regression model analysis to assess their prognostic value. It is suggested that improving survival rate for patients with SCLS can be obtained by using selective combined therapy. PMID- 2551472 TI - Respiratory paralysis as a presenting symptom in Japanese encephalitis--a case report. AB - In the literature there are rare reports on Japanese encephalitis (JE) mimicking poliomyelitis or presenting as respiratory paralysis. A case of JE presenting as respiratory paralysis is described. The 22-year-old male was well until five days earlier, when he experienced the acute onset of headache and fever. Four days after onset he developed dysphagia and respiratory difficulty, and a tracheotomy was performed. The following day he lapsed into semicoma and the respiration had to be maintained by an artificial ventilator. He regained consciousness by the 10th day after onset. He became able to comprehend questions and express himself by gestures. But the respiration was still paralytic, and assisted ventilation via a tracheotomy had to be continued. The case was clinically diagnosed as encephalitis. Bulbar poliomyelitis was initially considered due to the clinical features of aseptic meningitis, respiratory paralysis, pharyngeal paralysis with pooling secretions in the pharynx, tachycardia and elevated blood pressure. But poliovirus culture obtained from stool and appropriate serological studies were negative. The diagnosis of encephalitis due to JE virus was confirmed by hemagglutination inhibition test (acute phase HI titer 1:320; convalescent phase HI titer 1.1280). PMID- 2551473 TI - Combined occurrence of chyloperitoneum and chylothorax after surgery and chemotherapy for Wilms' tumor. AB - Chyloperitoneum is an extremely rare complication of abdominal surgery in children and a combined occurrence of chylothorax and chyloperitoneum after abdominal surgery has never been reported in children. Chylous ascites usually occurs as a result of operative trauma to the thoracic duct, cisterna chyli, or its tributaries. About one third of all patients with chylous ascites after retroperitoneal lymph node dissection also develop secondary chylothorax. Diaphragmatic defects have been shown to be responsible for the occurrence of chylothorax secondary to chyloperitoneum. Congenital diaphragmatic weakness may result in evagination of the peritoneum causing diaphragmatic blebs, the rupture of which results in the movement of the peritoneal fluid into the pleural cavity. In the authors' patient, the rent in the diaphragm that occurred during surgery was probably responsible for the chylothorax. The role of chemotherapy, if any, in the pathophysiology of this complication is unknown. Total parenteral nutrition (TPN) is a simple and effective treatment for postoperative chylous effusions. Surgical treatments such as abdominal exploration for the repair of leaking lymphatics and peritoneovenous shunt should be reserved for patients who fail TPN. PMID- 2551474 TI - Treatment of high-risk gestational trophoblastic disease with chemotherapy combinations containing cisplatin and etoposide. AB - The authors have treated 22 patients with high-risk gestational trophoblastic disease (GTD) by cisplatin-etoposide-containing combinations. Sixteen patients were treated with dactinomycin, platinum, and etoposide combination (APE regimen) and six patients had platinum and etoposide combination (PE regimen). Fourteen patients were treated for resistant or relapsing GTD after first-line therapy, and eight patients initially. All 22 patients were high risk according to the World Health Organization prognostic score values. Sustained complete remission was achieved in 19 patients (86%). All eight patients who received treatment as initial therapy were cured (100%) whereas only 11 patients were cured among the 14 patients who failed prior chemotherapy (78%). Hematologic and renal toxicities were limited and no treatment-related deaths occurred in this group of patients. Cisplatin and etoposide could be more widely used in chemotherapeutic combinations for high-risk gestational trophoblastic disease. PMID- 2551476 TI - A T-cell line with an unusual phenotype. AB - A rapidly proliferating T-cell line, HCD8, was derived from the peripheral blood lymphocytes of an apparently healthy individual during the course of a T-cell cloning experiment. This T-cell line expressed a very unusual phenotype: CD1+, CD2-, CD3+ (cytoplasmic), CD4-, CD5+, CD7+, CD8-, interleukin-2 receptor (IL-2 R) (p55)-, and T-cell antigen receptor (TCAR) alpha beta-. Assays for reverse transcriptase activity and for human T-lymphotropic retroviral sequences in the cellular DNA were negative, indicating that the cells were not transformed by human T-lymphotropic virus (HTLV)-I, HTLV-II, or human immunodeficiency virus (HIV)-I. Culturing the cells in the differentiation inducing agent 12-O tetradecanoyl phorbol 13-acetate induced an increased expression of CD3 but no other significant changes in T-cell markers. A small population of CD4-negative and CD8-negative T-lymphocytes exist in human peripheral blood and they exhibit natural killer (NK) and antibody-dependent cell-mediated cytotoxic (ADCC) activity. However, the authors' cell line failed to demonstrate such cytotoxic function. The TCAR gene rearrangement studies showed that both T gamma genes were rearranged while the T beta genes were in the germ line configuration and the T delta genes were deleted. HCD8 strongly expressed the antigens Leu M1 and Ki-1, markers detected only rarely on normal unstimulated human T-cells, but quite consistently found on Reed-Sternberg cells and cells of some large pleomorphic T cell lymphomas. HCD8 may be used to study the control of Leu M1 and Ki-1 expression in T-cells and it may provide some insight into the cellular origin of the above-mentioned lymphomas. PMID- 2551475 TI - Immunocompetence in lung cancer. Relationship to extent of tumor burden and histologic type. AB - In vitro assays of immunocompetence were done in 60 patients with differing extents of tumor load and various histologic types of lung cancer and were compared to values obtained for 60 normal controls. Profound alterations in monoclonal antibody-defined T-lymphocytes and circulating B-cells were seen. All patients showed impaired blastogenic response to the mitogens used with the exception of a normal response to pokeweed mitogen (PWM) in patients with localized disease. Increase levels of serum IgG were seen in patients with localized disease whereas high levels of IgA was seen in patients with more advanced disease. Distant metastases were associated with low IgM levels. All patients studied regardless of stage and histologic type had elevated levels of circulating immunocomplexes. These findings indicate gross immunologic abnormalities in these patients. PMID- 2551477 TI - Prognostic value of histologic grade nuclear components of Scarff-Bloom Richardson (SBR). An improved score modification based on a multivariate analysis of 1262 invasive ductal breast carcinomas. AB - We did a multivariate analysis of 1262 patients with operable, invasive ductal breast carcinoma to assess the prognostic value of the Scarff-Bloom-Richardson (SBR) histologic grading system. Nodal metastasis and SBR were the two most important factors for metastasis-free survival (MFS), P = 10-9 and P = 10-5, respectively, for total study time. In patients who were node negative, the SBR and International Union Against Cancer (UICC) stages were the most important for MFS (P = 4 X 10-4 and P = 0.03). In order to try to improve the SBR prognostic value, we first studied the three components of the SBR separately: ductoglandular differentiation proved the least predictive and nuclear pleomorphism and mitotic index the most predictive. A rearrangement of the two nuclear scores alone produced higher risk values and better risk separation of patient subpopulations than SBR, and eliminated the SBR from the multivariate model. This rearrangement, modified SBR (MSBR), defined five new risk subgroups with statistically different risk ratios for MFS (P = 3 X 10-8). SBR grade II (55% of patients) was separated into three MSBR groups significantly different according to MFS (P = 0.008). In the patients who were node negative, MSBR replaced the SBR and was the most important factor for prediction of relapse of MFS (P less than 0.00001). The MSBR is more accurate and predictive than the standard SBR grade and is particularly useful when the nodal status of the patient is negative or unknown. PMID- 2551478 TI - Neuroepithelial small cell carcinoma of the vagina. AB - We studied two patients with primary vaginal small cell carcinomas. Their tumors show histologic and ultrastructural neuroepithelial elements similar to those reported in the literature in the cervix and the lung. The clinical behavior of the tumors was aggressive in both patients. Ultrastructural study of small cell genital tract neoplasms is indicated to help classify this disease. Adjunctive therapy may be required for neoplasms of neuroepithelial origin. PMID- 2551479 TI - Peripheral primitive neuroectodermal tumor (peripheral neuroepithelioma) in children. A review of the St. Jude experience and controversies in diagnosis and management. AB - All patients diagnosed with primitive neuroectodermal tumor (PNET) and extraosseous Ewing's sarcoma in one institution between 1962 and 1987 were reviewed. Of the 26 cases studied, 16 had been diagnosed originally as PNETs, seven tumors were rediagnosed as PNET or EOE by histologic review, and three tumors had an original diagnosis of extraosseous Ewing's sarcoma. To determine whether these diagnoses determine a group of tumors with unique biologic behavior and identifiable pathologic characteristics, clinical and treatment response data were compiled, and electron microscopic and immunohistochemical studies were done for those patients with adequate samples. With combined modality therapy, this group achieved a substantially shorter disease control interval than patients with disseminated osseous Ewing's sarcoma or disseminated neuroblastoma--10.8 months versus 17 months and 16 months, respectively. The pattern of relapse and distant spread also differed among these tumor types. Immunohistochemical studies (for example, neuron-specific enolase and beta 2 microglobulin) were helpful in confirming the diagnosis but were not definitive in themselves. Tentative diagnostic criteria are proposed for use in studies designed to provide further information on the nature and treatment of PNET. Some of the controversies regarding diagnosis are discussed. The authors propose a uniform approach to treatment of extraosseous Ewing's sarcoma and PNET in order to try to clarify their relation. PMID- 2551480 TI - An ultrastructural study of benign and malignant breast epithelial cells. A search for tonofilaments. AB - 310 breast lesions were examined by electron microscopy. Tonofilaments were demonstrated in excretory epithelial cells of infiltrating ductal carcinoma (29%), infiltrating lobular carcinomas (27%), fibrocystic disease (18%) and in the normal breast (12%). Tonofilaments were also present in uncommon breast carcinomas, such as mucinous, papillary and medullary carcinomas. PMID- 2551481 TI - An ultrastructural evaluation of cell heterogeneity in invasive ductal carcinomas of the human breast. I. An in vivo study. AB - The present ultrastructural study has been undertaken in order to contribute to the problem of morphological heterogeneity in the ductal infiltrating carcinoma of the human breast. In spite of the well known topographical variability of scirrhous breast cancers, the comparative analysis of 12 primary tumours has brought to light some basic phenotypical expressions of the neoplastic cell population. The major observation is the occurrence of two main categories of cells, which are interpretable as the transformed counterparts of the dark and light lumenal cells of the normal mammary epithelium. The phenotypical identity of the two categories has been assessed by in vitro cultivation (parallel paper). Many aberrant morphological variants, attributable to the two cell types, were observed at the tumour-stroma interface. We have therefore suggested that the high level of morphological heterogeneity may, at least in part, be the result of stromal influences on the gene expression of the transformed cells. PMID- 2551482 TI - An ultrastructural evaluation of cell heterogeneity in invasive ductal carcinomas of the human breast. II. An in vitro study. AB - The ultrastructural characterization of the present continuous cell line, derived from a primary ductal infiltrating carcinoma (d.i.c.) of the human breast, has brought to light a remarkable morphological similarity with the original neoplastic cell population. A major parallelism is the permanent presence in culture of two categories of cells, exhibiting a strong isomorphism with the in vivo counterparts. The presence of well defined ultrastructural features, as duct like structures, microvillous projections, junctional complexes and intracytoplasmic crypts, is a further confirmation of the breast cancer origin of this line. The significance and perspective of these findings are discussed. PMID- 2551483 TI - The expression frequency of common fragile sites and genetic susceptibility to lung cancers. AB - The chromosomal aberration rate (including gap and break) and expression frequency of common fragile sites were examined in peripheral blood lymphocytes cultured with TC199 medium from 96 patients with lung cancers, 40 of their first degree relatives, and 45 normal control subjects. Both the chromosomal aberration rates and expression frequencies of common fragile sites observed in patients and their relatives were significantly higher than those in normal control subjects. About 60% of chromosomal aberrations were derived from the expression of common fragile sites either in the patients and their relatives or in the controls. The expression of fra(3)(p14) was most frequently observed, and the mean frequencies of its expression in patients and their relatives were significantly higher than in control subjects. It is suggested that common fragile sites might be unstable factors of the human genome, and their expression might be affected by some genetic factors, and they might play an important role in the genetic susceptibility to lung cancers. The significantly high expression of fra(3)(p14) in patients and their relatives may be related to the generation of the breakpoint at band 3p14 found in lung cancers. PMID- 2551484 TI - A human adult Wilms' tumor. Histologic, ultrastructural, and cytogenetic analysis. AB - The cytogenetic, histologic, and electron microscopic studies of an adult patient with Wilms' tumor are presented. Wilms' tumor (nephroblastoma) is a common renal tumor of childhood but is extremely rare in people over 15 years old. The histologic analysis of the patient's tumor, including both light and electron microscopic analysis, indicated that this tumor satisfies the histologic criteria for an adult Wilms' tumor, namely, blastemic cells that are immature renal parenchymal cells, embryonic tubular structures, and a scanty stromal component consisting of loosely arranged spindle cells. The tumor showed several ultrastructural features characteristic of adult Wilms' tumor, namely, markedly elongated mitochondria, autophagic vacuoles, and intracytoplasmic filaments. Karyotypic analysis was performed on the patient's peripheral leukocytes and tumor cells. The leukocytes showed no significant increase in gaps and breaks, and the patient appears to have a normal male karyotype. Some interesting chromosomal anomalies were observed in the cultured tumor cells: at least one chromosome 13, both chromosomes 22, and the X chromosome are missing, three markers are present, and there is a possible deletion of 12p. PMID- 2551485 TI - Detection of chromosome aberrations in interphase tumor nuclei by nonradioactive in situ hybridization. AB - In a blind study, chromosome aberrations in tumor cells were analyzed by conventional cytogenetic techniques (G banding) and nonradioactive in situ hybridization with chromosome-specific probes. The material was obtained directly from patients with hematologic diseases and from colon tumor derived cell lines. The cytogenetic data obtained with G banding were in accord with those obtained by in situ hybridization to metaphase chromosomes. Most importantly, in situ hybridization to interphase nuclei gave reliable results and even allowed detection of cell subpopulations that were not detected by analyzing metaphase chromosomes. Furthermore, in retrospect, even structural aberrations could be detected in interphase nuclei; abnormal cells with either an i(1q) or a translocation der(1)t(1;7) could be identified. Our results show that the application of in situ hybridization in combination with routine cytogenetic techniques offers significant advantages for cytogenetic analysis of solid tumors and hematologic malignancies. PMID- 2551486 TI - Preferential sites for viral integration on mammalian genome. AB - Chromosomal localization of human papillomavirus (HPV) 16 and 18 on human cervical carcinomas and epithelial cell lines obtained after HPV transfection has uncovered a nonrandom association of viral integration and specific genome sites. Fragile sites appear to be preferential targets for viral integration because of their structural and functional characteristics through which chromosomal anomalies, alterations in protooncogene activity, and gene amplification can occur. Individually or in association, such changes lead to the acquisition of an unlimited cell growth potential but not tumorigenicity. Genetic instability and uncontrolled cell division resulting from HPV integration increase the cell's susceptibility to other exogenous carcinogenic factors that may complete the process of neoplastic development. PMID- 2551487 TI - Patients with different lung cancers show normal expression of fra(3)(p14.2) in aphidicolin-treated lymphocyte cultures. AB - Among common fragile sites, fra(3)(p14.2) is the most expressed either spontaneously or after treatment with aphidicolin (APC) in lymphocyte cultures. Because recurrent chromosomal abnormalities involving the short arm of chromosome 3 in tumor tissue are present in various malignancies, including lung cancer, the induction of fra(3)(p14.2) elicited by APC was investigated with the aim of detecting possible interindividual polymorphism in its expression that might be relevant to predisposition toward cancer-related events. Thirty-four patients affected with various lung cancers (14 squamous cell carcinomas, 13 adenocarcinomas, and seven small cell carcinomas) and 14 controls (patients undergoing routine routine follow-up after coronary by-pass) were included in this study. The frequency of fra(3)(p14.2) expression was not significantly different among the patients grouped either by disease or by sex and age. It was estimated that fra(3)(p14.2) accounts for about 20% of total breakage in APC treated lymphocyte cultures from the general population. PMID- 2551488 TI - Lack of drug-induced DNA cross-links in chlorambucil-resistant Chinese hamster ovary cells. AB - Chlorambucil (CLB) is an alkylating agent commonly used in the treatment of several neoplastic disorders. The mechanisms underlying resistance to this drug are not fully defined. We used the DNA alkaline elution technique to study cross link formation in the wild type (K1) and a CLB-resistant (ChlR) Chinese hamster ovary cell line. [14C]CLB was used to measure drug uptake. The CLB-resistant cells were found to have negligible DNA cross-link formation compared to K1 cells at all time points tested. There was a correlation between the resistance to CLB and the decreased ability of resistant cells to form DNA cross-links. Results of drug uptake experiments excluded altered CLB accumulation as the basis for these findings. Assays of O6-alkylguanine transferase and topoisomerase. II provide evidence against a role of these enzymes in CLB resistance. These studies suggest that the mechanism of CLB cytotoxicity involves the formation of DNA cross-links. Reduced cross-link formation may confer resistance to CLB. PMID- 2551489 TI - Temperature dependence of adriamycin-induced DNA damage in L1210 cells. AB - We report alkaline elution experiments that reveal the temperature dependence of DNA lesions, both single-strand breaks and DNA-protein cross-links, in L1210 cells exposed to Adriamycin. DNA damage, which at 37 degrees C is equivalent to several hundred rads of ionizing radiation exposure, diminishes as the temperature of drug exposure is lowered. At all temperatures below about 15 degrees C no DNA damage is detectable in L1210 cells exposed to Adriamycin, even at relatively high doses. The low temperature inactivity is not due to a redistribution of intracellular drug since at both 37 and 0 degrees C there is a high concentration of Adriamycin in both nuclear and cytoplasmic locations. The temperature profile for DNA damage parallels the profile for cytotoxicity, i.e., at low temperature, the drug is completely inactive as a cytotoxic agent (P. Lane, P. Vichi, D. L. Bain, and T. R. Tritton, Cancer Res., 47:4038-4042, 1987). Thus, DNA breaks and cell kill appear to be correlated with one another. However, when we examined DNA lesions in nuclei isolated from L1210 cells we found that the low temperature inability to sustain Adriamycin-induced single-strand breaks or DNA-protein cross-links was absent. In nuclei, then, the drug can provoke DNA damage at low temperature, while in whole cells it cannot. Topoisomerase II, an enzyme implicated in catalyzing DNA lesions in cells exposed to intercalating agents, retains its catalytic activity both to unknot P4 DNA at 0 degrees C, and to be induced by drug to alter the release of pBR322 supercoils, so a low temperature inactivation of this enzyme cannot explain the results. We propose that intact L1210 cells have a regulatory factor which controls DNA damage, possibly through topoisomerase II, but which is lost when nuclei are isolated. PMID- 2551490 TI - Effect of dietary cellulose on cell proliferation and progression of 1,2 dimethylhydrazine-induced colon carcinogenesis in rats. AB - The effects of different levels of dietary cellulose on colonic crypt mitotic activity and colon carcinogenesis were studied in 190 male Sprague-Dawley rats. Rats were divided into groups and fed a basal fiber-free diet supplemented with either 0, 5, or 15% pure cellulose (w/w), for periods of 10 weeks (initiation stage) or 32 weeks (promotional stage). Half of the rats in each group were given weekly s.c. injections of 9.5 mg 1,2-dimethylhydrazine (the base) (DMH) for 8 weeks. Some of the rats were killed at 10 weeks while most were killed 22 weeks later. In some groups the dietary cellulose level was changed to a different level at 10 weeks. Food intake and body weight data showed that the rats within each experiment were isocalorically fed. There was a direct correlation between crypt height and the percentage of cellulose in the diet. Addition of 5 or 15% dietary cellulose during the initiation stage of carcinogenesis resulted in a significant increase in crypt height. Increasing dietary cellulose after the initiation stage (0 to 5% and 5 to 15%) or maintaining a high dietary cellulose level throughout both the initiation and promotional stages (15%) resulted in a significant increase in crypt height. A DMH-induced increase in mitotic activity that was observed during the initiation stage was no longer evident after the 22 week promotional stage. The significant DMH-induced increases in proliferative zone height and crypt height that were initially observed during the initiation stage were also observed after the 22-week promotional stage. These data indicate that the initial DMH-induced increases observed in proliferative zone height and crypt height are irreversible. Addition of 5 or 15% cellulose was found to suppress DMH-enhanced mitotic activity in the crypts of the descending colon during the initiation stage of carcinogenesis. This finding was correlated with a significantly lower incidence of adenocarcinomas in rats maintained on 5 or 15% cellulose throughout both the initiation and promotional stages. PMID- 2551491 TI - Abolition of mevinolin-induced growth inhibition in human fibroblasts following transformation by simian virus 40. AB - The basal level of the gene expression and the activity of 3-hydroxy-3 methylglutaryl coenzyme A (HMG CoA) reductase was higher in SV40-transformed human fibroblasts (90-VA VI) than in normal ones (HDF). In both these cell types mevinolin (25 microM) caused an 85-90% depression of HMG CoA reductase activity and of the incorporation of [3H]acetate into sterols. In HDF this was coupled to an efficient block of cell growth, whereas the growth of 90-VA VI was only slightly reduced by mevinolin. In HDF, mevinolin (25 microM) also abolished essentially all dilichol synthesis, as measured by incorporation of [3H]acetate. In contrast, dolichol synthesis remained unaltered, or was increased, in mevinolin-treated 90-VA VI. We suggest that these different responses of dolichol synthesis may depend on different substrate affinities of the rate-limiting enzyme in the dolichol pathway. However, if 90-VA VI was treated with 25 hydroxycholesterol (25-OH), an alternative inhibitor of HMG CoA reductase, the cellular growth as well as dolichol synthesis was significantly decreased. Since the inhibitory effect of 25OH on HMG CoA reductase activity did not exceed that of mevinolin, it seems that 25-OH, besides HMG CoA reductase, inhibits steps distal to HMG CoA reductase. This notion was further supported by the finding that addition of mevalonate did not prevent the 25-OH-induced growth inhibition. However, if dolichol was added along with 25-OH, the block was partially prevented, indicating that a critical level of de novo synthesis of dolichol for cellular growth. PMID- 2551492 TI - Reduced O6-methylguanine repair in fibroblast cultures from patients with lung cancer. AB - The activity of O6-methylguanine-DNA methyltransferase was determined in fibroblast cultures from 45 patients with lung cancer, 39 patients with cutaneous malignant melanoma, and 29 healthy controls. This enzyme is a critical parameter for the capacity to repair O6-methylguanine (O6-mGua) adducts in DNA, and a decreased activity might therefore be responsible for an enhanced susceptibility to cancer. The assay was performed with 8 x 10(6) fibroblasts which were homogenized and incubated with a known amount of O6-mGua containing DNA. The remaining substrate was determined fluorimetrically after high performance liquid chromatographic separation. O6-mGua repair was significantly reduced in lung cancer patients [6.64 +/- 4.32 (SD) pmol O6-methylguanine repaired/8 x 10(6) cells] as compared to healthy controls [10.35 +/- 5.42, P less than 0.0022] or patients with cutaneous malignant melanoma [10.83 +/- 6.66]. The lowest mean values were detected in a subgroup of 16 lung cancer patients with a tumor manifestation below 46 years of age (5.06 +/- 3.89). Fibroblasts from 4 patients with lung cancer had no detectable repair. We conclude that a reduced capacity to remove O6-mGua adducts may represent a further mechanism of individually enhanced lung cancer risk. PMID- 2551494 TI - Chromosomal changes in human primary testicular nonseminomatous germ cell tumors. AB - A cytogenetic analysis of 14 primary testicular nonseminomatous germ cell tumors has been carried out after short term tissue culture. The modal chromosome numbers ranged from 53 to 113, in agreement with flow cytometric determination of the DNA content of the tumors. At least one copy of an i(12p) was present in 12 tumors. Two tumors, however, lacked that marker. Some chromosomes are apparently overrepresented, whereas others are underrepresented, although some differences between seminomas and nonseminomas were noticed. PMID- 2551493 TI - Human IgG3 monoclonal antibody directed to an unbranched repeating type 2 chain (Gal beta 1----4GlcNAc beta 1----3Gal beta 1----4GlcNAc beta 1----3Gal beta 1--- R) which is highly expressed in colonic and hepatocellular carcinoma. AB - Previously established human monoclonal antibodies (MAbs) directed to carbohydrate antigens are essentially all IgM class, and show relatively low affinity and low reactivity at 37 degrees C. We report here the establishment of a human IgG3 MAb displaying high affinity antigen-binding activity at 37 degrees C and efficiently activating cellular cytotoxicity directed to human tumor cell lines expressing the polylactosamine antigen. The IgG3 MAb (MH21-134) reacted with the repeated unbranched polylactosamine structure Gal beta 1----4GlcNAc beta 1----3Gal beta 1----4GlcNAc beta 1----3Gal beta 1----R, i.e., nLc6, nLc8, etc., but did not react with sialyl 2----3 or 2----6 substituted derivatives at the terminal Gal. This specificity differs from that of several anti-i antibodies, or human anti-i-like MAbs which react with sialyl 2----3 substituted structures. Directly biotinylated MH21-134 antibody was used in immunohistochemical staining of 154 formalin-fixed, paraffin-embedded tissue sections to study distribution of the antigen. High incidence of positive staining was found in colon cancer (11/17; 65%) and hepatocellular carcinoma (8/12; 67%), followed by large cell and squamous cell carcinoma of lung cancer (10/13; 59%, and 14/26; 54%, respectively). TLC immunostaining of glycolipid extracts from a variety of tumor tissues showed the presence of nLc6 and/or nLc8 in over 50% of cases. The antigens nLc6 and nLc8 were found to be absent from normal colonic epithelia, kidney, and pancreas. Only a weak band corresponding to nLc8 and one corresponding to nLc6 were found in liver and spleen, although all these normal tissues, including gastrointestinal epithelia, lung, liver, spleen, erythrocytes, and lymphocytes, were essentially negative on immunohistology. However, the antigen was found to be highly expressed in myelocytes and weakly in bronchial glands of lung and pancreatic duct epithelia. Nevertheless, expression of unsubstituted, unbranched polylactosamine antigen could be an important basis for induction of humoral immune response against certain types of human cancer, despite its limited expression in normal cells. PMID- 2551495 TI - Regional distribution of human papillomavirus DNA and other risk factors for invasive cervical cancer in Panama. AB - A population-based national cancer registry has documented strikingly different regional incidence rates of cervical cancer in the Republic of Panama. Such regional differences in disease rates could represent regional differences in the occurrence of risk factors, in particular, human genital papillomaviruses (HPV). This study enrolled newly diagnosed invasive cancer patients in the Republic of Panama over an 18-mo period. Behavioral risk factors were measured by interviewing cases and matched controls. In addition, DNA extracted from biopsies of the cancers was tested for HPV sequences. Early age at first coitus, multiple pregnancies, and nonparticipation in Pap smear screening programs were significant risk factors for cervical cancer in this population. These factors and low levels of education occurred more frequently among women residing in regions with higher cancer rates than women residing in the region with lower cancer rates. HPV DNA was detected most frequently (70%) among cases from the region with the lowest cancer rate (30 of 100,000) and least frequent (54%) among cases where the cancer rate was the highest (51 of 100,000). The observations suggest that risk factors other than HPV contribute to the differences in cervical cancer rates among women residing in various regions of Panama. PMID- 2551496 TI - Decreased DNA interstrand cross-linking and cytotoxicity induced in human brain tumor cells by 1,3-bis(2-chloroethyl)-1-nitrosourea after in vitro reaction with glutathione. AB - Although both direct and glutathione S-transferase (GST)-catalyzed interactions between many electrophiles and GSH generally result in inactivation of the former, there are several reports of compounds whose electrophilic, alkylating, and cytotoxic activities are potentiated by GSH. This study investigates the effects of direct in vitro interaction between GSH and BCNU at physiological pH (7.2) and temperature (37 degrees C) and how this affects the cytotoxic and DNA cross-linking activity of 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) in target human malignant brain tumor cells. The kinetics and dose-response relationship of this interaction were determined by measuring residual GSH and residual BCNU cytotoxicity in aGSH/BCNU mixture over a 45-min period and at varying BCNU concentrations. The results demonstrate that reaction of BCNU with four times its molar concentration of GSH for 45 min significantly inactivates BCNU, as expressed by a 32% decrease in induction of cellular DNA cross-linking, a 21% increase in DNA synthesis, and a 15% increase in clonogenic survival of human brain tumor cells compared to incubates of BCNU alone. Equine liver (EL)-GST increased the inactivation of BCNU only slightly (insignificant at p = 0.05). These results suggest that, in contrast to agents such as the alkyl-N-nitro-N' nitrosoguanidines which become more potent alkylators after reacting with GSH, the 2-chloroethylnitrosoureas (CENUs) undergo inactivation by GSH. We propose that such interactions between GSH and the CENUs may constitute an important aspect of CENU metabolism and provide a potential means by which brain tumor cells can circumvent CENU toxicity and exhibit resistance to this class of agents. PMID- 2551497 TI - DNA topoisomerase II-mediated interaction of doxorubicin and daunorubicin congeners with DNA. AB - Three groups of doxorubicin and daunorubicin analogues, differing by their substituents on the chromophore and sugar moieties, were used in this study. The 3'-N-unsubstituted (Group 1), 3'-N-acyl (Group 2), and 3'-N-alkyl (Group 3) analogues were tested for: (a) in vivo antitumor activity and in vitro cytotoxicity; (b) cellular or tissue uptake and metabolic conversion; (c) strength of DNA intercalation; and (d) interaction with DNA topoisomerase II (topo-II). Compounds of Group 1 were cytotoxic, were strongly intercalative, and, except for those with C-14 side chain substitution, induced the formation of topo II-DNA cleavable complexes. As shown previously, esterolysis of C-14-acyl substituents was required to yield a metabolite which can interact with topo-II in the purified system. The C-14-substituted compounds of Group 2 and their C-14 unsubstituted metabolites were cytotoxic. These drugs were weak intercalators, and the C-14-unsubstituted cogeners induced cleavable complex formation in the purified system, but with reduced potency relative to doxorubicin. The type of the 3'-N-position substituent determined whether Group 3 analogues were cytotoxic and strong intercalators, or less active and nonintercalating. Although C-14 unsubstituted intercalators of Group 3 did not form cleavable complexes in the purified system, they were cytotoxic. The study shows that DNA intercalation is required but not sufficient for the activity by topo-II-targeted anthracyclines. In addition to the planar chromophore which is involved in intercalation, two other domains of the anthracycline molecule are important for the interaction with topo-II: (a) substitution of the C-14 position totally inhibits drug activity in the purified system, but enhances cytotoxicity by aiding drug uptake and presumably acting on other cellular targets; and (b) substitutions on the 3' N position of the sugar ring can, depending on the nature of the substituent, inhibit intercalation and/or topo-II-targeting activity. These findings may provide guidance for the synthesis and development of new active analogues. PMID- 2551498 TI - Examination of the role of the urokinase receptor in human colon cancer mediated laminin degradation. AB - The relevance of urokinase receptors to urokinase-mediated laminin degradation was investigated in cultured colon cancer. Six colon cancer cell lines degraded laminin in a plasminogen-dependent manner. The ability of the individual cell lines to cleave the glycoprotein correlated well (r2 = 0.9242) with the amount of urokinase recovered from the cell surface by a mild acid treatment. A radioreceptor assay indicated that colon cancer cells most active in degrading the laminin, possessed the largest number of urokinase receptors. Moreover, acid treatment which depletes the receptors of endogenous plasminogen activator augmented the specific binding of radioactive urokinase to the colon cancer cells by 12-200%. A cell line (HCT 116) which displayed 1.1 x 10(5) receptors/cell the majority of which were occupied with endogenous urokinase was selected and the effects of a urokinase receptor antagonist on laminin degradation determined. The peptide antagonist reduced laminin turnover by 60-80%. Morphological observations were consistent with these findings. Plasminogen-treated HCT 116 cells retracted from the culture dish and many cells were observed in the culture medium. This effect could be largely reversed by simultaneous treatment with the peptide antagonist. A poor correlation was found between laminin degradation and soluble urokinase (r2 = 0.1074). These data strongly argue for a role of the urokinase receptor in facilitating the action of the plasminogen activator in colon cancer. PMID- 2551500 TI - [Activation and inhibition of adrenergic effects on the coronary vessels of patients with different forms of angina pectoris]. AB - Experimental data in animals indicate that coronary vasoconstriction occurs following blockade of the beta-adrenergic receptors or alpha-receptors activation. The vasomotor effects of these maneuvers in man are unclear. Therefore we investigated whether and to which extent alpha-stimulation (cold pressor test: CPT) and beta-blockade (propranolol) cause coronary vasoconstriction; whether this effect involves the resistance arterioles as well as the large epicardial branches, and, within these, whether the normal and stenotic tracts are involved. Patterns in patients with effort angina were compared with those in patients with Prinzmetal angina. We studied 19 cases with classic and 15 cases with Prinzmetal angina. The systemic, pulmonary and coronary hemodynamics (pressure, flow and resistance) and the vasomotor pattern of normal and stenotic epicardial branches (quantitative angiography) were evaluated in the baseline condition, during CPT, after propranolol (5 mg iv) and during CPT repeated after propranolol. We observed that: changes of the coronary flow due to beta-blockade and to CPT are related to the variations of the myocardial oxygen consumption, induced by the inhibition and activation of adrenergic receptors and not to the concomitant vasomotor reaction of the stenotic vascular tract; beta blockade does not affect homogeneously the lumen of the stenotic lesions in effort angina and invariably increases the lumen in the Prinzmetal form; influences of CPT, in the absence as well as in the presence of beta-receptor blockade, on the lumen diameter of both normal vessels and stenotic lesions are minimal in either form of angina. PMID- 2551499 TI - Development of a transgenic mouse system for the analysis of stages in liver carcinogenesis using tissue-specific expression of SV40 large T-antigen controlled by regulatory elements of the human alpha-1-antitrypsin gene. AB - alpha-1-Antitrypsin (AAT) is the major antiprotease in human plasma; it is synthesized primarily in hepatocytes and to a lesser extent in several nonhepatic tissues. Under the control of regulatory elements of the human AAT gene, expression of SV40-large tumor antigen (T-ag) in transgenic mice occurred in the liver, stomach, pancreas, and kidney. Among seven founder transgenic animals, six developed liver carcinoma, four showed gastric neoplasia, and one developed pancreatic carcinoma. In three animals the kidneys showed glomerular or tubular epithelial hyperplasia but no malignancy. A stable transgenic line, 1812, was established. Members of this line reproducibly develop liver tumors by 10 weeks of age but do not exhibit any phenotypic changes in other tissues. Histological changes leading to liver tumor formation occurred with predictable kinetics and could be classified into four distinct stages: (a) embryonal/fetal stage, no recognizable histological changes; (b) newborn to 2 weeks of age, hyperplastic hepatocytes with reduced amounts of cytoplasm but no nuclear alterations; (c) between 3 and 8 weeks of age, diffuse liver cell dysplasia without observable tumor nodules; and (d) 8 weeks of age and thereafter, hepatocellular carcinomas in a background of liver dysplasia. Embryonic and newborn liver tissue showed uniform, high level expression of T-ag in the majority of hepatocytes by immunohistochemistry, whereas the dysplastic and tumoral stages were characterized by considerable variation in both the intensity of T-ag staining and the proportion of T-ag-positive cells. Immunoprecipitation analyses showed that T-ag was complexed with cellular protein p53 in all tumor samples. This study showed that SV40 T-ag expression in the liver resulted in cellular hyperplasia and dysplasia; additional event(s) apparently were required for progression to neoplasia. Those cooperating events occurred with predictable kinetics. This transgenic mouse system displays several similarities with human liver disease and provides a practical model for the study of separate steps in hepatocarcinogenesis. PMID- 2551501 TI - [Acute respiratory disease caused by coronaviruses]. AB - Serological evidence of coronavirus aetiology was found in 11 patients out of 218 persons examined (5%) in the period of 1986-1987, and in 23 cases out of 311 patients (7.4%) in the period of 1987-1988. Antibody titres with a minimum of four-time elevated values in paired sera using the ELISA method were considered conclusive. Coronaviruses 229E and OC43 were employed. During the first period, more persons were infected by coronavirus 229E, while in the second period, by coronavirus OC43. The disease was equally spread over all the months of the years. For example, in the 1987-1988 period, most infections occurred in February, when out of 60 examined patients, 9 were coronavirus positive (15%). PMID- 2551502 TI - [Occurrence of symptomatic liver porphyria in workers at risk for exposure to free SiO2 during subway construction]. AB - Symptomatic hepatic porphyria was followed up in workers exposed to the risk of free SiO2. This disease was found in 12 out of 998 workers engaged in the construction of the underground and exposed to SiO2 (group I), and in 12 at-risk persons out of 440 patients from different workplaces who were hospitalized for SiO2 exposure (group II). The average age of the workers of group I was 40.6 +/- 7.7, that of the group II 54.9 +/- 13.3. In both these high-risk groups, porphyria was found to be significantly more frequent than in the control group not exposed to free SiO2. The authors discuss the long-lasting effects of free SiO2 on the occurrence of symptomatic hepatic porphyria. PMID- 2551503 TI - Uptake and utilization of human polymorphonuclear leukocyte granule myeloperoxidase by mouse peritoneal macrophages. AB - Functional myeloperoxidase contained in granules of polymorphonuclear neutrophil leukocytes or in fixed whole cells can be endocytosed by mouse peritoneal macrophages. Acquired myeloperoxidase was distributed in what we considered to be the secondary lysosomal system and, following a phagocytic stimulation, was delivered to newly formed phagosomes containing the targets. PMID- 2551504 TI - Characterization of the follicles in the intermediate lobe of the pituitary gland of the Mongolian gerbil (Meriones unguiculatus). AB - The Mongolian gerbil (Meriones unguiculatus) contains abundant follicles throughout the intermediate lobe (IL) of the pituitary gland in the adult animal. The mode of follicle formation, the nature of the follicle building cells and the distribution of follicles were investigated in semithin sections of the gerbil IL. The sections were stained conventionally, or immunohistochemically with antibodies directed against alpha-melanocyte stimulating hormone (alpha-MSH). Follicular cells were constantly alpha-MSH-negative, and resembled the marginal cells lining the hypophyseal cleft with regard to their cytological and immunohistochemical properties. Moreover, follicular cells appeared to be derived from strands of marginal cells that regularly invaginated deep into the IL. Both follicular and marginal cells often made up cellular clusters. This process coincided with follicle formation and the generation or transport of the colloidal content found inside follicles and the hypophyseal cleft. Although the non-secretory cells of the IL obviously constituted one major source of pituitary colloid in the gerbil, alpha-MSH-positive secretory cells, which occasionally were found to be discharged into the cleft cavity, might contribute to the colloidal contents. PMID- 2551505 TI - The bovine papillomavirus E5 transforming protein can stimulate the transforming activity of EGF and CSF-1 receptors. AB - The bovine papillomavirus E5 transforming gene encodes a 44 amino acid protein product that is localized to cytoplasmic membranes, including the plasma membrane. We now report that E5 can cooperate with human EGF receptors and with human CSF-1 receptors to induce cellular transformation of NIH 3T3 cells. Cooperation occurred in the absence of receptor stimulation by ligand, and it was further augmented by treatment with ligand. Cooperation was not seen between E5 and either c-fes or c-src. The cooperation between E5 and high levels of EGF receptors was associated with inhibition of receptor degradation and persistence of activated receptors on the cell surface. We conclude that E5 may enhance the receptor activity via inhibition of receptor down-modulation. PMID- 2551508 TI - Identification of Alu transposition in human lung carcinoma cells: a correction. PMID- 2551506 TI - Lysophosphatidate-induced cell proliferation: identification and dissection of signaling pathways mediated by G proteins. AB - Lysophosphatidate (LPA), the simplest natural phospholipid, is highly mitogenic for quiescent fibroblasts. LPA-induced cell proliferation is not dependent on other mitogens and is blocked by pertussis toxin. LPA initiates at least three separate signaling cascades: activation of a pertussis toxin-insensitive G protein mediating phosphoinositide hydrolysis with subsequent Ca2+ mobilization and stimulation of protein kinase C; release of arachidonic acid in a GTP dependent manner, but independent of prior phosphoinositide hydrolysis; and activation of a pertussis toxin-sensitive Gi protein mediating inhibition of adenylate cyclase. The peptide bradykinin mimics LPA in inducing the first two responses but fails to activate Gi and to stimulate DNA synthesis. Our data suggest that the mitogenic action of LPA occurs through Gi or a related pertussis toxin substrate and that the phosphoinositide hydrolysis-protein kinase C pathway is neither required nor sufficient, by itself, for mitogenesis. The results further suggest that LPA or LPA-like phospholipids may have a novel role in G protein-mediated signal transduction. PMID- 2551507 TI - Phosphorylation of middle T by pp60c-src: a switch for binding of phosphatidylinositol 3-kinase and optimal tumorigenesis. AB - Substitution of phenylalanine for tyrosine 315 of the polyoma virus middle T (mT) protein lowers the incidence and limits the spectrum of tumors induced following inoculation of the virus into newborn mice. This substitution removes the major site of phosphorylation by pp60c-src without altering the ability of mT to associate with or to activate pp60c-src. The mutant mT fails to show binding of a phosphatidylinositol 3-kinase (Ptdlns 3-kinase) activity that is normally present in wild-type mT complexes. Furthermore, an anti-peptide antiserum that specifically recognizes mT lacking phosphate at tyrosine 315 precipitates binary (mT-pp60c-src) but not ternary (mT-pp60c-src-Ptdlns 3-kinase) complexes from wild type infected cell extracts. Reprecipitation with either anti-pp60c-src or anti mT serum brings down ternary complexes containing mT phosphorylated on tyrosine 315. Phosphorylation of mT by pp60c-src in vivo is therefore a critical event for binding of Ptdlns 3-kinase and for expression of the full tumorigenic potential of the virus. PMID- 2551509 TI - Defective signal transduction in CD4-CD8- T cells of lpr mice. AB - Mice homozygous for the lpr gene develop a lymphoproliferative disorder due to expansion of a subset of CD4-CD8- T cells. Triggering of the T-cell receptor in these lpr T cells does not lead to translocation of protein kinase C or phosphorylation of CD3, interleukin-2 production, or proliferation, whereas a combination of phorbol ester and calcium ionophore does. Stimulation with concanavalin A or anti-CD3 induces phosphoinositide hydrolysis. The rise in inositol bisphosphate, inositol triphosphate, and inositol tetrakisphosphate, identified by HPLC, is similar in +/+ and lpr T cells. The concentration of cytoplasmic free calcium ([Ca2+]i), however, under basal and stimulated conditions is significantly lower in lpr T cells. The lower basal [Ca2+]i may explain why induction of proliferation with phorbol ester and calcium ionophore requires a higher concentration of ionophore in these cells than in normal T cells. The lower [Ca2+]i obtained on stimulation may contribute to the activation defect of CD4-CD8- lpr T cells. PMID- 2551510 TI - Partial purification and characterization of the soluble glutathione transferase isoenzymes from cultured Hep G2 cells. AB - Hep G2 cells, an established cell line derived from a human hepatoma, were mass cultured in a cell factory for the isolation of glutathione transferase isoenzymes. These were enriched by affinity chromatography and separated in an anionic and a cationic fraction. They were partially characterized by different kinetic and inhibition parameters. Three different subunits were observed. The results were compared with human liver data. It is concluded that Hep G2 cells can be considered as a valuable alternative tool for in vitro research of human liver phenomena, especially when toxicological interactions are investigated. PMID- 2551511 TI - Proliferating, transformed keratinocytes cultured under low Ca2+ conditions exhibit high-affinity epidermal growth factor receptors. AB - In a previously published report (Exp. Cell. Res. 161:421 (1985] we have demonstrated that cultured normal and transformed keratinocytes exhibit two classes of EGF binding sites after growth under normal Ca2+ conditions but only low-affinity binding sites after growth under low Ca2+ conditions. Here we demonstrate the presence of high-affinity binding sites in transformed keratinocytes grown under low Ca2+ conditions, using a specific monoclonal anti EGF-receptor antibody. PMID- 2551512 TI - [A nosocomial epidemic caused by adenovirus type 3 at a clinic for children and adolescents]. AB - The authors describe an epidemic of acute viral respiratory diseases with a nosocomial character at a clinic for children and adolescents, caused by adenovirus type 3. Of 80 exposed children 26 fell ill (attack rate 32.5%). The disease took a clinical course typical for adenovirus type 3. The authors discuss the problem of respiratory infections with a viral aetiology at paediatric departments and their possible prevention. PMID- 2551513 TI - [A minor epidemic of pertussis cough in Brno]. AB - In the course of six months the authors investigated 32 children aged 1-32 months with pertussoid cough. This number included one child where pertussis was confirmed in a 25-month-old properly vaccinated child and parapertussis in an 8 month-old incompletely vaccinated infant. In the remaining children by cultivation or serological examination a different aetiology was found (RS virus, adenovirus, parainfluenza 2, M. pneumoniae, H. influenzae, Branhamella catarrhalis). PMID- 2551514 TI - [A medium available in Czechoslovakia for the culture of Entamoeba histolytica and Giardia intestinalis]. AB - From nutritive media produced in the CSSR a modification of Diamond's medium TYI S-33 was prepared which ensures the long-term axenic cultivation of Entamoeba histolytica and Giardia intestinalis. Details of the cultivation technique are also presented. PMID- 2551515 TI - [Epidemic keratoconjunctivitis. Laboratory study of patients infected with adenovirus type 8]. AB - The authors tried to detect, using different methods, the causal agent of the disease in eight patients with epidemic keratoconjunctivitis. From the conjunctival sacs of one ambulatory patient and six of seven patients hospitalized on account of other eye diseases they isolated 11 strains of adenovirus type 8 on cell cultures from human embryonic lungs. Using direct and indirect ELISA, they detected adenovirus antigen in four of seven examined smears from the conjunctival sac without previous cultivation. In smears from the nasopharynx they did not detect adenoviruses by either of the two methods. Antibodies were assessed by micromodifications of the virus neutralizing test, by the complement fixation reaction and ELISA. A significant rise of antibodies against adenovirus type 8 and adenovirus hexone resp. was found in different six of eight examined pairs of sera. The combination of these three serological methods made it possible to confirm adenovirus infection in all eight patients. PMID- 2551516 TI - [Biochemical requirements in tocolysis]. AB - By a specific bond of 3H-dihydroalprenolol the number (Bmax) and characteristic (Kd) of sympathetic beta-receptors in cell membranes of the myometrium and lymphocytes of peripheral blood was assessed. The authors examined six tissue specimens taken during planned termination of pregnancy during the 38th-40th week by primary Caesarean section parallel with a peripheral blood sample. In healthy women even at the end of pregnancy the myometrium contains ample amounts of specific beta-receptors. As compared with the myometrium, the lymphocytes of the peripheral blood contain beta-adrenergic receptors of the same type as the myometrium but in much smaller amounts. So far it was not possible to show a correlation between the number of sympathetic beta-receptors in the membranes of the myometrium and of lymphocytes in the peripheral blood. PMID- 2551517 TI - [Late results in the replacement of auditory ossicles with silastic prostheses]. AB - Silastic prostheses of the middle ear proved useful in the reconstruction of the chain of auditory ossicles. Following an observation period of 6 months to 6 years they led in type PORP in a group of 49 subjects to early functional positive results in 78% and late positive results in 71%. The authors tested type TORP in 56 patients with a 57% success as regards early and late results. They used pistons only in obliterated fossae of the fenestra ovalis or in case of thick immobile plates. Despite this they achieved excellent and good early and late auditory gains in 58%. Of 105 prostheses type PORP and TORP administered during the entire observation period rejection occurred only in four subjects, i. e. in 3.8%. They inserted a strip of fascia between the cap of the prosthesis and tympanic membrane. PMID- 2551518 TI - [The pathophysiologic basis for the incorporation of 99mTc-pyrophosphate into myocardial infarct]. AB - The investigation studied the relation of 99mTc-pyrophosphate incorporation (99mTc-PYP) into experimental, 48-hours old myocardial infarction in dogs to tissue vascular supply and to the extent of necrotic tissue. The experimental myocardial infarction was induced in five animals during an operation by the ligation RIVA. The myocardial blood supply was measured in tissue samples, taken from transverse sections of the infarction by means of 86Rb captation. The extent of myocytolysis was measured by depletion of tissue creatine kinase (CK). In the subendocardial layer of the infarction it became obvious that 99mTc-PYP incorporation was proportional to the decrease of tissue blood supply. No relation between the extent of necrosis and the incorporation of the radioactive chemical was demonstrated. The accumulation of 99mTc-PYP requires the necrosis to be present, but its extent apparently does not influence the intensity of incorporation. In the subepicardial infarction layer there was neither a relation of the radioactive chemical cumulation to the blood flow, nor to the extent of the necrosis proved. PMID- 2551519 TI - [Treatment of 100 patients with small cell lung cancer]. AB - 100 patients with small cell lung cancer were treated in Peking Union Medical College Hospital from 1963 through 1987. The five year survival rate in patients received combined treatment (resection + chemotherapy+radiotherapy, resection+chemotherapy or resection+radiotherapy,) was 18.52% (5/27). It was better than patients received nonsurgical treatment (chemotherapy,radiotherapy or chemotherapy+radiotherapy, 58 patients). And also it was better than patients received surgical resection only (11 patients). This advantage was especially marked in patients with lesion of IIIa stage. The five year survival rate in this group was 26.31%(5/19). It was suggested that combined method (resection+chemotherapy+radiotherapy) is the first choice for the treatment of small cell lung cancer. PMID- 2551520 TI - pH-adjustment of 2-chloroprocaine quickens the onset of epidural anaesthesia. AB - The effect of pH-adjustment of three per cent 2-chloroprocaine (2-CP, Nesacaine MPF) on the onset, duration, and spread of epidural analgesia and anaesthesia was studied in patients undergoing lower extremity surgery. Forty ASA physical status I and II patients were randomized to two groups. In a double-blinded fashion, patients in both groups received an epidural injection of 15 ml of local anaesthetic (LA) solution via a Tuohy needle at the L3-4 interspace. Local anaesthesia for Group I was prepared by adding 3 mEq NaHCO3 to 27 ml three per cent 2-CP and for Group II was prepared by adding 3 ml 0.9 per cent NaCl to 27 ml three per cent 2-CP. Both solutions contained epinephrine (1:200,000). The pH of commercially prepared Nesacaine MPF was 3.19 +/- 0.02. The pH of the solutions used for Group I and Group II patients were 7.32 +/- 0.01 and 3.27 +/- 0.02, respectively. Times to analgesia and anaesthesia at the L2 dermatome were significantly decreased in Group I patients by 2.5 and 6.6 minutes, respectively. Likewise, pH-adjustment accelerated the attainment of maximum level of block by 2.8 min. No statistical differences were found between groups in the maximum level of epidural block, or in time to 2-segment regression. No precipitation of LA was observed in pH-adjusted solutions of 2-CP after 24 hours. We recommend the use of pH-adjusted three per cent 2-CP (Nesacaine MPF) to accelerate the onset of epidural block. PMID- 2551521 TI - Combination chemotherapy with cisplatin and etoposide associated with radiotherapy in the treatment of small-cell lung cancer. AB - A total of 52 consecutive, previously untreated patients with small-cell lung cancer (SCLC) were scheduled to receive six cycles of a combination of etoposide (75 mg/m2 per day) and cisplatin (20 mg/m2 per day), each cycle given over 5 consecutive days. In all, 28 patients had extensive disease (ED) and 24, limited disease (LD). After three cycles of chemotherapy, all responding patients were given chest radiotherapy (RT) (45 Gy in two split courses and 30 Gy in LD and ED, respectively); only patients with LD who achieved complete remission (CR) after three cycles of chemotherapy were given prophylactic brain irradiation (30 Gy). In the 51 evaluable patients, the overall response rate was 90%, with a 31% CR and a 59% partial remission (PR) rate. In LD and ED patients, 57% and 11% CR rates and 30% and 82% PR rates were noted, respectively. Myelosuppression was the most frequently observed toxicity. The median duration of response was 12 months in LD (range, 3-41 + months) and 7 months (range, 2-12 months) in ED; the median survival was 15 months in LD and 9.3 months in ED, respectively. In all 30% of LD patients are alive and well at a minimal follow-up of 18 months. This trial confirms the activity of the cisplatin-etoposide combination in SCLC. PMID- 2551522 TI - Arachidonic acid potentiates superoxide anion radical production by murine peritoneal macrophages stimulated with tumor promoters. AB - The role of arachidonic acid (AA) metabolism in the stimulation of oxygen radical production by murine peritoneal macrophages treated with tumor promoters was assessed. In vivo administration of the phospholipase A2 inhibitor dibromacetophenone, the anti-inflammatory steroid fluocinolone acetonide or the lipoxygenase inhibitor nordihydroguiaretic acid just prior to i.p. injection of phorbol-12-myristate-13-acetate (PMA, 100 ng) into unmanipulated CD-1 female mice resulted in a dose-dependent decrease in the number of peritoneal exudate cells (PEC) producing superoxide anion radical (O2) as assessed by the reduction of nitroblue tetrazolium, i.e. the formation of formazan-positive PEC. The cycloxygenase inhibitor indomethacin had no effect on the number of formazan positive PEC caused by PMA treatment. The ability of PMA, phorbol-12,13 dibutyrate mezerein, phorbol-12,13-diacetate and 4-O-Me-PMA to stimulate the production of oxygen radicals by murine peritoneal macrophages correlated with their ability to stimulate the release of [3H]AA equivalents from the macrophages. The calcium ionophore A23187 which stimulated significant [3H]AA equivalent release did not stimulate superoxide anion radical production by the macrophages. PMA administered i.p. to SENCAR mice increased the number of formazan-positive PEC 4-to 5-fold compared with similarly treated C57BL/6 mice. PMA also stimulated the release of twice the amount of [3H]AA equivalents from peritoneal macrophages from SENCAR mice compared with that released by macrophages from C57BL/6 mice. The addition of low concentrations of AA (1-10 microM) in vitro to casein-elicited murine peritoneal macrophages treated with low concentrations of PMA (1 ng/ml) resulted in a 2-fold potentiation of the amount of superoxide anion radical produced compared with PMA treatment alone as assessed by the reduction of cytochrome c. These results demonstrate that AA and/or a lipoxygenase product can potentiate the production of oxygen radicals by murine peritoneal macrophages treated with tumor promoters. PMID- 2551523 TI - Specific high frequency rearrangements induced by MNNG in SV40-infected human keratinocytes. AB - In order to study carcinogen-induced changes in integrated viral sequences clonal sublines of SV40-transformed human keratinocytes were exposed to N-methyl-N' nitro-N-nitrosoguanidine (MNNG) at sublethal concentrations ranging up to 10 micrograms/ml for a period of 15 min and then examined by Southern blot hybridization using full-length SV40 DNA as a probe. Of the clonal sublines tested, one (AG34) was found to exhibit certain consistent, dose-dependent changes at 4 days post-treatment: (i) loss of at least two EcoRI fragments of approximately 4.4 and 3 kb, with the concomitant appearance of two bands migrating between 1.8 and 2.3 kb; and (ii) loss of a 1.5-kb fragment and the appearance of 2, 3.8 and 5 kb fragments in KpnI digests. Minor variable changes in restriction patterns were seen at 24 h post-treatment but consistent and pronounced effects were observed only at 4 days post-treatment. The altered restriction fragment patterns indicate that MNNG caused highly specific rearrangements in some subset of the DNA sequences associated with the integrated SV40 sequences in human epithelial cells. This differs from what has been found for SV40 sequences in other cell lines where amplification has been reported. These results suggest that (i) the site of SV40 integration may determine the response of integrated SV40 segments after carcinogen treatment, and (ii) carcinogen treatment can result in the induction of a common genetic event throughout the entire population of exposed cells. Genomic libraries created in a lambda phage vector have been used to isolate BamHI fragments containing SV40 sequences. Isolates have been found which exhibit EcoRI and KpnI restriction patterns consistent with the polymorphisms displayed in Southern blots. PMID- 2551524 TI - Protection against cadmium toxicity and enzyme inhibition by dithiothreitol. AB - In the present in vivo studies the alterations in cation transporting enzymes of the brain, kidney and liver tissues were assessed at intervals between 0 to 48 h after a single, acute (10 mg kg-1, i.p.) dose of cadmium (Cd). The inhibition of Na+-K+-ATPase during the first 24 h does not parallel the changes in K+-PNPPase suggesting differential effects on phosphorylation and dephosphorylation steps of the overall ATPase reaction. Between 30 min to 2 h the inhibition in enzyme activity was steep (27 per cent in brain, 54 per cent in liver) followed by a rapid reversal between 2-6 h. This critical period may correspond to the time of induction of metallothionein. This enzyme reversal was followed by a significant decrease in Na+-K+ ATPase (40-68 per cent) and K+-PNPPase (44-60 per cent) between 24 to 48 h. A similar pattern was observed in Ca2+-ATPase in all the three tissues. A 33 per cent mortality was observed in rats after 48 h of cadmium challenge. Administration of dithiothreitol (DTT, 20 mg kg-1, i.p.) to CdCl2 pretreated rats at 24 h resulted in mortality reduced from 33 per cent to 0 and reversal in the inhibition of Na+-K+-ATPase in brain and kidney and Ca2+-ATPase in brain. Since protection of brain and kidney enzymes by DTT paralleled its protection against Cd toxicity, their inhibition by Cd may, in part, constitute the biochemical basis of Cd toxicity. PMID- 2551525 TI - Role of intracellular Na+ in Ca2+ overload and depressed recovery of ventricular function of reperfused ischemic rat hearts. Possible involvement of H+-Na+ and Na+-Ca2+ exchange. AB - The roles of H+-Na+ and Na+-Ca2+ exchange in the depression of ventricular function were studied in the reperfused isolated ischemic rat heart. Zero-flow global ischemia was induced for either 15 or 30 minutes and was followed by 30 minutes of aerobic reperfusion. Intracellular Na+ (Na+i) and 45Ca2+ uptake were measured during ischemia and reperfusion. Accumulation of Na+i was modified by prior glycogen depletion and by treatment with amiloride, a H+-Na+ exchange inhibitor, or monensin, a Na+ ionophore. Na+i rose continuously during ischemia and rapidly during the first two minutes of reperfusion. The larger inhibitory effect of amiloride and preischemic glycogen depletion was on Na+i accumulation during reperfusion; this finding suggests that the uptake occurs by H+-Na+ exchange. Reduction of Na+i accumulation by glycogen depletion was associated with less lactate and, presumably, H+ production and accumulation during ischemia. The rapid increase in Na+i during early reperfusion may reflect the readjustment of the low intracellular pH resulting from ischemia. The level of Na+i at the end of ischemia and especially after two minutes of reperfusion were linearly correlated with 45Ca2+ uptake and depression of ventricular function during subsequent reperfusion. This highly significant correlation between Na+i and 45Ca2+ uptake when Na+i was varied by several independent procedures, including monensin, strongly suggests that reperfusion 45Ca2+ uptake occurs at least in part by Na+-Ca2+ exchange. The rate of 45Ca2+ uptake during reperfusion was linearly and highly significantly correlated with elevation of diastolic pressure, reduced developed pressure, and decreased recovery of ventricular function. The data strongly support a mechanism of ischemic cell damage that involves excessive production and accumulation of H+ during ischemia that exchanges for extracellular Na+ during ischemia and rapidly during the first few minutes of reperfusion. Increased Na+i then causes excessive 45Ca2+ uptake and depressed recovery of cellular functions with continued reperfusion. Increased levels of Na+i may be a major event that couples a decreased intracellular pH during ischemia to excessive 45Ca2+ uptake and depressed recovery of cellular function with reperfusion. PMID- 2551526 TI - Enhanced sensitivity of heart cells to adenosine and up-regulation of receptor number after treatment of guinea pigs with theophylline. AB - Experiments were carried out in hearts from guinea pigs that were fed either the adenosine receptor antagonist theophylline (0.6 mg/ml) or no drug. The A1 adenosine receptor radioligand [125I]aminobenzyladenosine bound to a single affinity class of receptors in heart cell membranes from control animals with Bmax and KD of 18.3 +/- 1.0 fmol/mg protein and 3.7 +/- 0.6 nM, respectively (n = 8). Heart cell membranes from animals fed theophylline for 2, 7, and 14 days bound the radioligand with about the same affinity, but the number of binding sites was significantly increased (p less than 0.01) to 30.6 +/- 1.7 (n = 3), 30.0 +/- 0.8 (n = 3), and 27.3 +/- 2.9 (n = 4), respectively. Nearly identical results were obtained with membranes prepared from enzymatically dispersed ventricular myocytes. Fourteen days of theophylline treatment also produced a small increase (12%, p less than 0.01) in the number of binding sites in membranes derived from cerebral cortexes. Isolated ventricular myocytes prepared from animals fed no drug or theophylline for 7 days were used to determine the effect of adenosine on 20 nM isoproterenol-stimulated calcium current (ICa) measured by the whole-cell patch-clamp technique. Adenosine reduced isoproterenol stimulated ICa without affecting the activation or inactivation kinetics of the current; ICa density was reduced less by 5 microM adenosine in cells from control (25 +/- 3 to 21 +/- 3 microA/microF) than in cells from theophylline-fed animals (26 +/- 5 to 17 +/- 2 microA/microF). Although a high concentration (0.5 mM) of adenosine abolished isoproterenol-stimulated ICa in cells from control or theophylline-fed animals, the IC50 for adenosine was sixfold less in cells derived from theophylline-fed animals than in cells from control animals (4.6 +/- 0.6 microM and 28.3 +/- 1.4 microM, respectively, p less than 0.01). In contrast, the increase in ICa in response to isoproterenol alone and the potency of acetylcholine to antagonize this effect of isoproterenol were the same in both groups of cells. A maximally effective concentration of R phenylisopropyladenosine (0.1 mM) inhibited isoproterenol-stimulated cyclic AMP accumulation less in cardiomyocytes from control than from theophylline-fed animals (28.7 +/- 1.8% vs. 42.0 +/- 4.2%, p less than 0.05). In summary, exposure of the myocardium to theophylline increases the number of adenosine receptors and the effects of receptor occupancy by agonists. These findings imply that the endogenous concentration of adenosine is high enough in the normoxic guinea pig heart to chronically maintain adenosine receptors in a partially down-regulated state. PMID- 2551527 TI - F(ab')2 fragments of anti-Mo1 (904) monoclonal antibodies do not prevent myocardial stunning. AB - To determine if inhibition of leukocyte adhesion and aggregation could improve postischemic ventricular dysfunction ("stunning"), a monoclonal antibody (904) that binds to the adhesion-promoting Mo1 glycoprotein on the cell surface of leukocytes was administered intravenously (0.5 mg/kg) to open-chest dogs before a 15-minute coronary occlusion. Ultrasonic crystals placed in ischemic and control myocardium were used to measure systolic wall thickening during a 15-minute occlusion of the left anterior descending artery and for 3 hours after reperfusion. Myocardial blood flow was measured with tracer-labeled microspheres before occlusion, after 10 minutes of occlusion, 3 minutes of reperfusion, and at 1 and 3 hours after reperfusion. Six animals receiving anti-Mo1 antibody had antibody excess demonstrated with immunofluorescence techniques at 5 minutes and 3 hours of reperfusion; this finding indicated saturation of binding sites. Five animals served as controls and received an antibody (murine immunoglobulin G) that does not influence neutrophils. The two groups did not differ hemodynamically during ischemia and reperfusion. Risk areas and myocardial blood flow were also not significantly different between the two groups. The main parameter used to define regional myocardial stunning, percentage systolic wall thickening in the ischemic/reperfused area, did not differ significantly between the two groups. Specimens from nonischemic myocardium were compared with ischemic specimens for myeloperoxidase content. There were no significant differences within or between groups. These data indicate that the anti-Mo1 monoclonal antibody (904) is not effective in improving the profound myocardial dysfunction that persists for 3 hours of reperfusion after 15 minutes of ischemia. PMID- 2551528 TI - Ca2+ uptake by cardiac sarcoplasmic reticulum from patients with idiopathic dilated cardiomyopathy. AB - We measured Ca2+ uptake by sarcoplasmic reticulum prepared from left ventricular myocardium obtained from six nonfailing human hearts and nine excised hearts from patients with class IV idiopathic dilated cardiomyopathy. Ca2+ uptake had a Vmax of 593 +/- 82 nmol/mg-min, a K0.5 of 0.68 +/- 0.07 microM, and an nHill of 1.7 +/ 0.1 in the nonfailing hearts. The corresponding values in the excised failing hearts were 593 +/- 36 nmol/mg-min, 0.63 +/- 0.03 microM, and 1.6 +/- 0.1. The beta-receptor density in crude sarcolemma prepared from left ventricular myocardium was 110.0 +/- 15.3 fmol/mg in the unmatched donors and 52.1 +/- 4.5 fmol/mg in the excised failing hearts. These results suggest that abnormal Ca2+ handling in idiopathic dilated cardiomyopathy in humans is not the result of any intrinsic alteration of Ca2+ uptake by sarcoplasmic reticulum. PMID- 2551529 TI - One hour of myocardial ischemia decreases the activity of the stimulatory guanine nucleotide regulatory protein Gs. AB - The effect of 1 hour of myocardial ischemia on the function of the stimulatory guanine-nucleotide-binding protein Gs was examined. This study follows our recent finding that myocardial ischemia increases the density of beta-adrenoreceptors in a conscious canine model while having the opposite effect on the activity of adenylate cyclase. Coronary artery occlusion was induced in five conscious dogs and verified by measurement of blood flow using the Doppler and microsphere techniques. Alterations in the level and function of Gs were examined in sarcolemmal membranes prepared from ischemic and nonischemic regions of the left ventricle. After 1 hour of coronary artery occlusion, the functional activity of sarcolemmal Gs, as determined by reconstitution with cyc- membranes, decreased by 27 +/- 7% in the ischemic zone. Cholera toxin labeling performed in parallel with the reconstitution studies demonstrated a similar decrease of 28 +/- 7%. This was associated with decreases in basal activity and decreases in adenylate cyclase activity stimulated by GTP, GTP plus isoproterenol, sodium fluoride, and forskolin. Thus, a defect distal to the beta-adrenoreceptor occurs in the transduction of adrenergic signals to the heart as a consequence of 1 hour of ischemia. PMID- 2551530 TI - Inhibition of sarcolemmal carbon-centered free radical formation by propranolol. AB - The mechanism of propranolol-inhibited sarcolemmal membrane lipid peroxidation was investigated by electron spin resonance spin-trapping technique using 5,5 dimethyl-1-pyrroline-N-oxide (DMPO) and 2-methyl-2-nitrosopropane (MNP). Highly purified canine myocytic sarcolemma were peroxidized by a superoxide-driven (from dihydroxyfumarate) and Fe3+-catalyzed free radical-generating system. Hydroxyl radicals (.OH), identified by electron spin resonance signals as DMPO-OH adducts, were generated in the aqueous phase. Propranolol up to 500 microM did not effectively reduce the intensity of the DMPO-OH adducts. When the sarcolemma were incubated with MNP before the addition of free radicals, MNP adducts characteristic of carbon-centered radicals were produced. Pretreatment of the membranes with propranolol (3-100 microM) decreased the intensity of the MNP adducts in a log concentration-dependent manner; the EC50 is about 7 microM. D- and L-propranolol were found equally effective. When protein-depleted sarcolemmal lipids were similarly incubated with MNP and the free radical system, identical MNP adducts were observed; this finding suggests that the adducts were lipid derived products arising from lipid peroxidation. Furthermore, their formation was also inhibited by propranolol pretreatment. Since propranolol is not an effective scavenger of oxygen radicals in the aqueous phase, the data suggest that the antiperoxidative effect of propranolol is due to its lipophilic interaction with the membrane and thus subsequent interruption of the free radical chain reactions. PMID- 2551531 TI - Thyrotropin-releasing hormone given intrathecally to the rat increases arterial pressure and heart rate. AB - In view of evidence implicating thyrotropin-releasing hormone (TRH) as a chemical mediator of synaptic transmission onto spinal sympathetic neurons, this peptide was administered intrathecally, in a dose of 6.5 nmol, at the T9 and T2 spinal levels in the anesthetized rat. At the lower thoracic level TRH increased arterial pressure and heart rate; these effects peaked at 4-7 minutes and decayed over the next 15-20 minutes. At the upper thoracic level the pressor and cardioacceleratory responses were roughly similar in time course but were smaller in magnitude. Hexamethonium (10 mg/kg i.v.) was tested on the responses from the lower thoracic level; both pressor and cardioacceleratory responses persisted after hexamethonium pretreatment. In addition, intravenous administration of 6.5 nmol of TRH failed to alter arterial pressure or heart rate, suggesting that the effects produced by the intrathecal administration of TRH were due to an action of the peptide in the spinal cord. The results also indicate that the pressor effect and the increase in heart rate may be mediated in the sympathetic ganglia at least partly via nonnicotinic transmission. Our results provide physiological support for the possibility that TRH is a chemical mediator of synaptic transmission onto sympathetic preganglionic neurons. This study indicates that the functional sympathetic pathways utilizing TRH as a chemical mediator include those regulating arterial pressure and heart rate. PMID- 2551533 TI - Absence of effects of cyclosporine on myocardial lymphocyte subsets in Coxsackievirus B3 myocarditis in the aviremic stage. AB - To test the therapeutic efficacy of immunosuppression with cyclosporine upon the aviremic stage of coxsackievirus B3 (CB3) myocarditis, 2-week-old BALB/c mice were inoculated with 3 x 10(2) plaque-forming units of CB3, and the effects of cyclosporine on peripheral, splenic, and myocardial lymphocyte subsets were investigated. Cyclosporine, 25 mg/kg/day, was administered subcutaneously daily on days 10-31 (experiment 1) and days 30-51 (experiment 2). Treated groups were compared with infected controls for each experiment. In experiment 1, the survival rate of the cyclosporine-treated group was low (17/25 vs. 24/25, p less than 0.05). The severity of myocardial lesions and the distribution of lymphocyte subsets in myocardium and spleen on days 15-18 did not differ between treated and control groups; on the other hand, the percentages of peripheral Thy 1.2+ (pan T) and L3T4+ (activated helper T) cells on days 15-18 were decreased in the treated group, and those of B, Lyt 1+ (helper/inducer T), and Lyt 2+ (suppressor/cytotoxic T) subsets did not differ significantly. Notably, myocardial interleukin-2 receptor (IL-2R) positive cells, through which cyclosporine is considered to act, were scarce in both groups. In experiment 2, survival rates of two groups did not differ (treated, 32/34; untreated, 34/34; p = NS). The severity of myocardial lesions and the distribution of splenic lymphocyte subsets on days 35-38 also were not different between two groups. The percentages of peripheral lymphocyte subsets (Thy 1.2+ and L3T4+) were decreased in the treated group; those of B, Lyt 1+, and Lyt 2+ subsets did not differ significantly. In experiments 1 and 2, the thymus/body weight ratio in the treated groups was smaller than in the untreated group, but the spleen/body weight ratio in the treated group did not differ from the untreated group; histologically, medullary cellular depletion was evident in the thymus, not in the spleen, of the treated groups. We conclude that cyclosporine failed to change the distribution of lymphocyte subsets in the spleen as well as in the myocardium in CB3 myocarditis although it had effects on the peripheral blood and thymus, which may account for the higher mortality in the treated groups. The absence of beneficial effects of cyclosporine upon the CB3-infected myocardium may be related to the paucity of cyclosporine-sensitive cells (IL-2R, L3T4, and Lyt 2 positive cells) in the myocardium. PMID- 2551532 TI - Rabbit polymorphonuclear neutrophils elicit endothelium-dependent contraction in vascular smooth muscle. AB - The present studies were designed to investigate the interaction between activated polymorphonuclear neutrophils (PMNs) and endothelial regulation of vascular smooth muscle function. Rabbit peritoneal PMNs (4 x 10(3)-4 x 10(5) cells/ml) added to muscle bath chambers containing phenylephrine-precontracted rabbit isolated aortic rings produced no effect on vascular tone. However, when PMNs were activated with the chemotactic peptide, f-met-leu-phe (0.1 microM), PMNs produced concentration-dependent vascular contraction, which was dependent on the presence of the endothelium. Aortic rings denuded of endothelium were unaffected by activated PMNs. Superoxide dismutase (100 units/ml) treatment of tissues blocked completely PMN-induced vascular contraction, whereas mannitol (20 mM) had no significant effect on PMN-induced vascular contraction. Pyrogallol (a generator of superoxide anion) produced a response that was similar to that observed with activated PMNs. Superoxide anion production was measured separately, and the time of peak rate of superoxide anion production corresponded to the time of the maximal vascular contractile responses. Activated PMNs added to vascular tissues undergoing endothelium-dependent relaxation mediated by either acetylcholine or A23187 produced a reversal of vascular relaxation. Furthermore, activated PMNs did not have any effect on endothelium-independent vascular relaxation produced by either isoproterenol or nitroglycerin. The present investigation reveals that activated PMNs can release superoxide anion and produce endothelium-dependent contraction. The endothelium-dependent contraction may be the result of superoxide anion inactivation of endothelium derived relaxing factor. PMID- 2551534 TI - Changes in phosphoinositide turnover in isolated guinea pig hearts stimulated with isoproterenol. AB - The incorporation of 32Pi into phospholamban, troponin I, phosphatidylinositols, and inositol trisphosphates was studied in Langendorff-perfused guinea pig hearts stimulated with isoproterenol. Hearts were perfused with Krebs-Henseleit buffer containing [32P]Pk and freeze-clamped at different times during the positive inotropic response. Exposure of the hearts to 0.1 microM isoproterenol for up to 1 minute was associated with significant (up to threefold) increases in phospholamban and troponin I phosphorylation, but there was no significant increase in 32P incorporation into phospholipids. However, longer exposure (2 minutes or more) to isoproterenol was associated with increases in the degree of 32P labeling of phosphatidylinositols and phosphatidic acid. Examination of 32P labeling of inositol trisphosphates in the same hearts revealed that the radioactivity associated with these compounds decreased with time. The decreases were significant at times of exposure of 2 minutes or longer to beta-adrenergic stimulation. The tissue levels of the inositol 1,4,5-trisphosphate isoform were also measured in hearts perfused with isoproterenol for 3 minutes, and they were found to be significantly lower compared with values obtained in control hearts. The effects of isoproterenol on 32P incorporation into phospholipids and proteins were observed in the presence of prazosin, and they were completely abolished by the beta-receptor blocker propranolol. Examination of the phosphoinositide specific phospholipase C activity in the perfused hearts revealed that isoproterenol stimulation was associated with a decrease in the membrane associated enzymatic activity at physiological calcium concentrations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551535 TI - Opiate receptor inhibition improves the blunted baroreflex function in conscious dogs with right-sided congestive heart failure. AB - The endogenous opiate system is activated in congestive heart failure. because endogenous opioids are known to depress the baroreflex function, we conducted studies to determine whether the increased endogenous opioids play a role in causing the reduced baroreflex function that occurs in heart failure. Right-sided congestive heart failure was produced in 16 dogs by tricuspid avulsion and progressive pulmonary artery constriction. Seven sham-operated dogs were included for comparison. Baroreflex function was measured in the conscious dogs after pretreatment with either normal saline or an opiate-receptor antagonist by bolus administration of phenylephrine. The slope of the regression line relating systolic blood pressure to cardiac cycle (R-R) interval was taken as an index of baroreflex sensitivity. Plasma beta-endorphin was elevated in the dogs with heart failure (15.3 +/- 2.5 pmol/l) compared with the sham-operated dogs (4.2 +/- 0.4 pmol/l, p less than 0.001). The dogs with heart failure also exhibited a reduced baroreflex sensitivity (3.84 +/- 0.19 msec/mm Hg) after saline pretreatment when compared with the sham-operated dogs (10.86 +/- 1.20 msec/mm Hg, p less than 0.001). Administration of naloxone hydrochloride increased the baroreflex sensitivity of dogs with heart failure to 5.16 +/- 0.26 msec/mm Hg (p less than 0.01) but produced no significant effects in sham-operated dogs (11.36 +/- 1.42 msec/mm Hg). To further study the site of action for the effect of naloxone, we measured baroreflex sensitivity in the dogs with heart failure after pretreatment with naloxonazine, a selective mu-receptor antagonist, with ICI 154,129, a selective delta-receptor antagonist, or with naloxone methobromide, a quaternary analogue of naloxone that does not penetrate the blood-brain barrier.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551536 TI - The enigma of primary Raynaud's disease. PMID- 2551537 TI - Treatment of acute venous thromboembolism with low molecular weight heparin (Fragmin). Results of a double-blind randomized study. AB - We performed a prospective, randomized, double-blind trial in 194 unselected patients to determine the safety and efficacy of low molecular weight heparin (Fragmin) compared with standard heparin as the initial treatment of acute venous thromboembolism. Ninety-eight patients received continuous intravenous heparin, and 96 patients received Fragmin for 5-10 days. Doses were adjusted to maintain anti-Xa levels between 0.3 and 0.6 unit/ml for patients with a high risk for a bleeding complication and between 0.4 and 0.9 unit/ml for patients with a low risk for bleeding. Treatment was stopped when a therapeutic level of anticoagulation (International Normalized Ratio greater than 3.5) was reached with coumarins. Thirteen patients in the heparin group and 10 patients in the Fragmin group had a major bleeding complication. The incidence of major and minor bleeding complications combined decreased from 48.9% to 38.5% (95% confidence interval for the difference, -3.5% to +24.2%), corresponding with a relative bleeding risk reduction of 21.2%. There were no significant differences in efficacy as defined by new high-probability defects on repeat ventilation perfusion scintigraphy of the lung in 80 patients: six of 46 patients in the heparin group and 3 of 34 patients in the Fragmin group had new defects (95% confidence interval for the difference, -9.4% to +17.8%). We conclude that low molecular weight heparin (Fragmin) given in adjusted, continuous, and intravenous doses is safe and effective as initial treatment of acute venous thromboembolism compared with heparin. There is a trend in risk reduction for bleeding in favor of low molecular weight heparin, a trend, however, that is smaller than expected compared with animal studies. PMID- 2551539 TI - Developmental changes in synaptic properties in hippocampus of neonatal rats. AB - The properties of synaptic responses in area CA1 of hippocampus were analyzed in slices prepared from 7-9 and 12-15 day old neonate rats. As expected from earlier work, only slices of two-week-old animals showed a consistent degree of long-term potentiation (LTP) in response to patterned high frequency stimulation. Several other synaptic properties were found to change during this developmental period. Inhibitory responses were absent in 7-9 day old but not in 12-15 day old neonates. Paired-pulse facilitation and the calcium sensitivity of postsynaptic responses were considerably reduced in 7-9 as compared to 12-15 day old rats. However, phorbol esters and 4-aminopyridine treatment still produced a strong facilitation of field potentials. The N-methyl-D-aspartate (NMDA) component of responses to single pulse stimulation in low magnesium medium was found to be larger in slices of 7-9 than 12-15 day old or adult animals. At the two time periods examined, trains of high frequency stimulation applied in the presence of regular magnesium elicited an NMDA dependent response. It is concluded that the differences in synaptic properties observed between 7-9 and 12-15 day old neonates may not account for the absence of LTP in the younger animals. PMID- 2551538 TI - Acute electrophysiologic effects of sodium administration in dogs treated with O desmethyl encainide. AB - Conduction slowing is the major in vivo effect of sodium channel blocking drugs. Although this action may promote arrhythmia suppression, apparently paradoxical arrhythmia aggravation does occur. The latter outcome is most frequently seen during treatment with the class IC agents such as encainide or flecainide, which are potent depressors of conduction even at usual plasma concentrations and heart rates. Anecdotal reports in patients with such drug toxicity have suggested a beneficial effect of sodium lactate or NaHCO3 administration. The purpose of this study, therefore, was to examine the changes induced by sodium loading on the electrophysiologic properties of the canine ventricle pretreated with a class IC drug. Thirty dogs received loading and maintenance infusions of O-desmethyl encainide (ODE), an encainide metabolite that as a sodium channel blocker is approximately 10 times more potent than the parent drug. Interventions were administered during the maintenance phase when stable plasma ODE concentrations of 448 +/- 68 (SEM) ng/ml were present, and QRS was prolonged from 62 +/- 1 to 89 +/- 2 msec, and HV was prolonged from 28 +/- 1 to 50 +/- 1 msec. NaHCO3 (5 meq/kg during 1 minute) shortened QRS from 92 +/- 6 to 76 +/- 3 msec and shortened HV from 44 +/- 3 to 37 +/- 3 msec within 10 minutes (both p less than 0.01). NaHCO3 also significantly prolonged endocardial monophasic action potential duration from 231 +/- 22 to 272 +/- 33 msec and decreased serum [K+] from 3.8 +/- 0.2 to 3.0 +/- 0.2 meq/l, but it did not alter plasma ODE concentration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551540 TI - Immunoradiometric assay for alpha gamma- and gamma gamma-enolase (neuron-specific enolase), with use of monoclonal antibodies and magnetizable polymer particles. AB - Monoclonal antibodies were raised against neuron-specific enolase, gamma gamma enolase, and used in an immunoradiometric assay (IRMA), with mono-disperse magnetizable particles as the solid phase. The assay's sensitivity was 0.4 microgram/L and the interassay coefficient of variation was less than 5% in the working range from 0.4 to 170 micrograms/L. Compared with our radioimmunoassay based on polyclonal antibodies, the incubation time is shorter, and precision and sensitivity are improved. The IRMA also improved detection of neuron-specific enolase in sera from patients with lung cancer without a concomitant change in measured enolase in the reference population. The better sensitivity of the IRMA results from its ability to measure alpha gamma- and gamma gamma-enolase with equal response. Ninety percent of the small-cell lung carcinoma patients (36 of 40) had increased values before treatment, compared with 7% of non-small-cell lung carcinoma patients (8 of 114). PMID- 2551541 TI - Oxalate is overestimated in alkaline urines collected during administration of bicarbonate with no specimen pH adjustment. AB - We compared measurements of daily urine oxalate excretion in urines collected at the prevailing urine pH with measurements of urine oxalate excretion in urines collected into 20 mL of 6 mol/L HCl. We studied eight healthy adults fed constant diets. Urines were collected during control conditions and, in each subject, during the administration of NaCl, KCl, NaHCO3, or KHCO3, 90 mmol/day. Daily urine oxalate excretion calculated for collections made in acid averaged 271 (SD 79) mumol/day and did not vary with any of the salt supplements. When urines were collected at ambient urine pH (average 5.94, SD 0.23) during control conditions, and during the administration of NaCl or KCl, urine oxalate excretion averaged 263 (SD 88) mumol/day, a value not different from that for collections in acid. However, when urine was collected with no pH adjustment during NaHCO3 or KHCO3 administration (average pH 6.90, SD 0.14), apparent urine oxalate excretion averaged 398 (SD 132) mumol/day, significantly (P less than 0.025) exceeding the mean observed when urines were collected in acid. Moreover, the percentage increase in apparent oxalate excretion increased with urinary pH. These observations reinforce recommendations that urine specimens for measurement of oxalate be collected in acid to avoid the increase in apparent oxalate content that occurs during collection of alkaline urines. This increase presumably results from the well-known in vitro nonenzymatic conversion of ascorbate to oxalate. PMID- 2551542 TI - Interaction between pyridine nucleotide coenzymes and heme proteins as a possible source of error in assay of activities of coenzyme-linked enzyme. AB - The ultraviolet absorbance spectra of pyridine nucleotide coenzymes change in the presence of heme-containing proteins. The positions of each of the two main absorbance peaks of NADH are shifted progressively towards shorter wavelengths in the presence of increasing concentrations of hemoglobin, and the third peak, at 220 nm, disappears altogether. Similar changes are seen in the spectra of NAD+ and NADPH, and similar effects on these spectra are produced by myoglobin and cytochrome c, but not by comparable concentrations of albumin. The spectral shifts are generally accompanied by a decreased peak height. This finding may help explain problems reported by previous workers in the measurement of the activity of enzymes such as transketolase or lactate dehydrogenase in erythrocyte hemolysates. Errors may be considerable if allowance is not made for this effect, especially if the concentration of heme protein in the spectrophotometer cuvette much exceeds 1 g/L. The interaction appears to indicate some form of bonding, occurring generally between pyridine nucleotide coenzymes and the heme group in proteins. We relate the findings to measurement of activities of pyridine nucleotide-linked enzymes in erythrocyte lysates and in plasma containing myoglobin after muscle breakdown. PMID- 2551543 TI - Biochemical contribution to diagnosis and study of a new case of D-glyceric acidemia/aciduria. AB - During organic acid screening by gas chromatography/mass spectrometry, we detected a large peak corresponding to glyceric acid in a patient's urine sample. The D(+) configuration was demonstrated by a polarimetric method and by enzymatic stereospecificity of D-glycerate dehydrogenase (EC 1.1.1.29). We biochemically investigated this fifth reported case of D-glyceric acidemia. In our patient, loading tests with L-serine and fructose led to an increase of D-glyceric acid in both plasma and urine. Determination of other metabolites involved in D-glycerate metabolism revealed no abnormality in any sample examined. After comparing all our results with those of the preceding observations described in the literature, we suggest a possible enzymatic defect located on one of the metabolic pathways shared by fructose and L-serine, possibly at the level of hepatic D-glycerate kinase (EC 2.7.1.31). Nevertheless, a primary defect of L-serine catabolism cannot be entirely excluded. PMID- 2551545 TI - Yohimbine and aldosterone responsiveness to angiotensin II or corticotrophin in normal subjects. AB - In normal man the sympathetic nervous system exerts an inhibitory influence on aldosterone responsiveness to angiotensin II. The possible role of alpha-2 adrenoceptors was assessed by studying the changes of plasma aldosterone during an angiotensin II infusion at the dose of 1, 2, 5 and 10 ng/kg. min or after corticotrophin infusion, 0.25 mg, in 8 normal subjects before and after treatment with the selective alpha-2 adrenoceptor antagonist, yohimbine, at a maximal dosage of 60 mg daily. Yohimbine did not modify blood pressure, body weight, the supine levels of angiotensin II, renin and aldosterone, the pressor response to angiotensin II and the correlation relating plasma aldosterone to plasma angiotensin II obtained during infusion studies. These findings suggest that the inhibitory influence of the sympathetic nervous system on aldosterone responsiveness to angiotensin II. PMID- 2551544 TI - Endogenous inhibition of red blood cell Na,K-ATPase in essential and pregnancy induced hypertension. AB - Digoxin-like inhibitors of Na,K-ATPase have been implicated in the pathophysiology of essential (EH) and pregnancy-induced hypertension (PIH). A technique that enhances dissociation of digoxin from red blood cells (RBC) was used to displace endogenous digoxin-like substances from RBCs. RBC membranes were preincubated in Na and ATP (Release) or Na,K,Mg and ATP (Retention) prior to measuring ATPase activity. Groups studied were: 39 men with EH and 34 controls plus 10 women with PIH and 17 normotensive controls. All displayed similar increases in Na,K-ATPase activity (24.0 +/- 7.9%) following Release. Plasma digoxin immunoreactivity (DI) was measured in pregnant women, m = 0.25 +/- 0.07 ng/ml. No DI was detected in nonpregnant women, but RBCs from these women demonstrated the same increase in Na,K-ATPase activity after Release. The 24% increase in activity achieved by Na and ATP preincubation can be reversed by adding K and Mg to the Release suspension. However, after RBC-bound digoxin is displaced by Release preincubation, addition of K and Mg cannot promote renewed binding and pump inhibition. Thus, the observed endogenous inhibition is not due to displacement of a digoxin-like substance but probably is related to alteration of the enzyme-membrane interaction. Furthermore, even though pregnant women demonstrate DI, an inhibitory substance with digoxin-like binding could not be recognized using this technique. PMID- 2551546 TI - Current strategies for atherosclerosis and lowering cholesterol. AB - Atherosclerosis is a multifactorial disorder in which nutritional factors are closely related to a number of the manifestations of the disease. Sounder nutritional principles might therefore prevent atherosclerosis itself or its major complications. Optimising nutrients would diminish monocyte adhesion to endothelium, suppress platelet aggregation, prevent oxidative modification of lipoproteins, minimise activation of coagulant factors and reduce the flux of cholesterol and of atherogenic lipoproteins through plasma. Optimising the mix of dietary fatty acids should have the highest priority. Emphasis should be placed on those fatty acids, such as those derived from fish, which modify several cardiovascular risk factors, including plasma lipids, blood pressure and coagulation factors. Reduction of risk factors is also achieved but to a lesser extent by reducing total fat intake, reducing dietary cholesterol, optimising the mix of starches and fibre and increasing plant foods generally. A developing strategy is to identify genetic predisposition to environmentally induced hyperlipidaemia, since most forms of hyperlipidaemia reflect interactions between genetic and dietary factors. PMID- 2551548 TI - Effects of atrial natriuretic factor on cyclic GMP content in the rat aortic smooth muscle: studies on the role of membrane Na+,K+-ATPase. AB - 1. These studies were conducted to determine whether preservation of the functional integrity of the membrane, Na+,K+-stimulated ATPase is essential for the atrial natriuretic factor (r-ANF-8-33) to enhance guanosine 3',5' monophosphate (cGMP) content in the rat aortic smooth muscle. In freshly dissected rat aortic tissues, levels of cGMP were estimated using radioimmunoassay. 2. ANF (0.1 mumol/L in Krebs-Henseleit media) produced significant elevation in cGMP levels in the aortic smooth muscle when compared with that incubated in the control media, whereas suppression of Na+-pump with ouabain (1.0 mmol/L) and/or K+-free media did not produce any significant changes in the basal cGMP level; these two experimental manoeuvres did not prevent enhancement of cGMP by ANF. 3. Incubation of the tissues in the media containing ouabain plus vasoconstrictor concentrations of norepinephrine (0.3 mumol/L) also did not alter basal cGMP levels and did not prevent the ability of ANF to elevate cGMP. 4. These studies demonstrate that the antagonism by ouabain, of vasorelaxant effects of ANF (as reported in the literature) are not due to the prevention of the ability of ANF to enhance cGMP levels in the arterial smooth muscle. It is proposed that such an antagonism may be related to the actions of ouabain and ANF on diverse, and perhaps independent, mechanisms which affect Ca2+ fluxes across the cell membrane. PMID- 2551547 TI - Diet, alcohol and hypertension. AB - Obesity, diet and alcohol consumption constitute major environmental determinations of blood pressure elevation. The long term setting of blood pressure in response to these factors will be determined by genetic susceptibility, and interactions with effects of physical fitness and smoking. Dietary changes which independently influence both atherosclerosis and hypertension are likely to be of greatest value in helping to control morbidity and mortality from hypertensive cardiovascular disease. Recommendations should focus on diets low in total and saturated fat intake and high in fruit and vegetables, containing potassium and fibre, coupled with weight control, alcohol moderation to less than two drinks per day in drinkers and regular physical exercise. Sodium restriction will help lower blood pressure in older hypertensives in particular. The role of dietary calcium or fish oils in blood pressure regulation is still uncertain. Dietary and related recommendations on smoking and exercise should be 'first line' treatment in mild hypertensives, and complimentary to therapy in all patients requiring drugs. PMID- 2551549 TI - Plasma vasopressin and atrial natriuretic factor during haemorrhage: influence of cardiac and aortic receptors. AB - 1. The changes in plasma concentrations of immunoreactive vasopressin (iVP) and atrial natriuretic factor (iANF) in response to haemorrhage (10-30% blood volume) were measured in 10 anaesthetized rabbits before and after cardiac receptor denervation (vagal nerve section). Carotid sinus pressure was maintained constant (60 mmHg) to eliminate any changing input from carotid baroreceptors. 2. Haemorrhage increased iVP before and after vagal nerve section indicating that withdrawal of input from aortic baroreceptors may have contributed to the increase in iVP. 3. Section of the vagus nerves attenuated the iVP response to haemorrhage. 4. There was no correlation between release of iVP and iANF. 5. Haemorrhage decreased iAF before and after vagal nerve section. Section of the vagus nerves increased iANF. Plasma iANF was highly correlated with atrial pressure and mean arterial pressure suggesting iANF release was secondary to changes in cardiac haemodynamics. PMID- 2551550 TI - Progesterone antagonizes development but not maintenance of ACTH-induced hypertension in sheep. AB - 1. This study investigated the effect of progesterone, which, under certain circumstances, can antagonize both the mineralocorticoid and glucocorticoid activities of steroid hormones, on the development and maintenance of adrenocorticotrophic hormone (ACTH)-induced hypertension in conscious sheep. 2. Progesterone (500 mg/day) alone, for 5 days, had no effect on blood pressure, but increased urinary Na excretion by 38 +/- 10 mmol/day (P less than 0.05) during the first 24 h. 3. Infusion of ACTH (5 micrograms/kg per day), alone, for 3 days, increased arterial pressure by 21 +/- 2 mmHg (P less than 0.001) associated with hypernatraemia, hypokalaemia, urinary Na retention, and increased fasting plasma glucose concentration. 4. Progesterone (500 mg/day) concurrently with ACTH blocked the rise in mean arterial pressure and the mineralocorticoid (urinary Na retention) but not the glucocorticoid (increase in plasma glucose concentration) effects associated with ACTH administration. 5. Progesterone (500 and 1000 mg/day) failed to reverse the hypertension and hypokalaemia in sheep pretreated for 3 days with ACTH. 6. Thus, progesterone blocked the onset but did not affect established ACTH hypertension. The mechanism by which progesterone blocked the development of ACTH hypertension appears to be related to the ability of progesterone to block the essential mineralocorticoid component of the adrenocortical steroids involved in the development of ACTH hypertension. PMID- 2551551 TI - Reactivity of anti-neutrophil cytoplasmic autoantibodies with HL-60 cells. AB - Anti-neutrophil cytoplasmic autoantibodies (ANCA) react with antigens in the cytoplasm of neutrophils and monocytes, and are found in the blood of patients with necrotizing vasculitis and pauci-immune necrotizing and crescentic glomerulonephritis. The standard techniques for ANCA analysis use human polymorphonuclear leukocytes (PMN) as a source for antigen. A comparison was made between ANCA reactivity with human PMN and HL-60 cells. Fifty-five ANCA-positive and 17 ANCA-negative sera were reacted with HL-60 cells in an indirect immunofluorescence microscopy assay to assess the ability of HL-60 cells to distinguish between positive and negative reactivity and between the two major types of ANCA. The HL-60 cell method agreed with a PMN method in all but one instance. In an enzyme immunoassay, HL-60 primary granules could be used to detect ANCA. Evidence is also presented that the perinuclear staining of HL-60 cells and neutrophils by ANCA with anti-myeloperoxidase specificity is caused by the nucleophilic properties of myeloperoxidase. Reactivity with HL-60 cells further elucidates the antigen specificity of both types of ANCA and indicates that HL-60 cells are a suitable tissue culture cell line for investigating the pathobiology of ANCA. PMID- 2551552 TI - Antigen-specific suppressor T lymphocytes in man. AB - The cellular signals that lead to activation of suppressor T cells (Ts) as opposed to cytotoxic T cells (CTL) are unknown. This review describes an in vitro suppressor-induction system developed by us to characterize interactions among various T cells leading to the development of antigen-specific suppression. In this system, antigen-specific CD4+ inducer T cells are first activated with antigen-presenting cells (APC). Antigen-primed CD4+ inducer blasts are then cultured with fresh autologous CD8+ T cells in the absence of the priming antigen. CD8+ T cells isolated from this culture suppress the proliferative response of autologous CD4+ T cells to the priming antigen only. The activated CD8+ Ts lyse neither APC nor antigen-primed CD4+ inducer T cells and can be distinguished from their CD8+CD28+ CTL counterpart by their lack of expression of the CD28 molecule. Furthermore, the ability to induce CD8+ Ts is restricted to antigen-primed CD4+CD29+CD45R-p80+ (Leu8+) T cells. Antibody-mediated inhibition experiments suggest the involvement of CD3/TCR and class I MHC molecules on the surface of CD4+ inducer T cells and the CD2, CD3/TCR, CD8, and CD11a/CD18 molecules on the surface of CD8+ Ts during both the induction and the effector phase of Ts function. Furthermore, compatibility at the class I MHC genes between CD8+ Ts and CD4+ antigen-reactive T cells is required for effective suppression of CD4+ T cells. Together, these results suggest that human antigen-specific CD8+ Ts employ the TCR complex to recognize TCR and class I MHC molecules on the surface of autologous CD4+ inducer T cells during the induction and effector phases of Ts function, and the apparent antigen specificity of suppression reflects specificity for antigen receptors on CD4+ antigen-reactive T cells. This may be a common mechanism by which antigen-specific suppression is accomplished. PMID- 2551554 TI - A pedigree of cervical stenosis, brachydactyly, syndactyly, and hyperopia. AB - Cervical myelopathy due to developmental cervical canal stenosis occurred in a 13 year-old boy. The patient's father and aunt also had an abnormally small cervical canal, although both were asymptomatic. The patient and his family had many congenital anomalies including hereditary brachydactyly, syndactyly, and hyperopia. The association of these anomalies seems not to have been previously reported in the literature. PMID- 2551553 TI - Connexin32, a gap junction protein, is a persistent oogenetic product through preimplantation development of the mouse. AB - Gap junctions appear de novo during compaction in the eight-cell stage of mouse development. This is a critical event in the life of the embryo, because gap junctional intercellular communication is an essential requirement for maintaining compaction and, hence, for development of the blastocyst. Recently, a family of genes encoding gap junction proteins (connexins) has been identified and cloned, and we have taken advantage of the availability of antibodies and cDNA probes to investigate the expression of these genes in early development. We found that a protein with antigenic and size similarity to the "liver" gap junction protein, connexin32, is present throughout preimplantation development from the zygote through the late morula. Connexin32 mRNA, however, could not be detected in any preimplantation stage. This, and the presence of connexin32 in zygotes before activation of embryonic transcription, leads us to conclude that this protein is inherited as an oogenetic product that persists well beyond the transition from the oogenetic to embryonic program of gene expression. Furthermore, we found that mRNA for another gap junction protein, connexin43, is fairly abundant in preimplantation embryos. We conclude that it is more likely connexin43, and not connexin32, that is used to assemble new connexons as the level of intercellular coupling increases after compaction. PMID- 2551555 TI - [Paralytic ileus and atonic bladder in a case of mitochondrial encephalomyopathy- electrophysiological, chemical and pathological study with evidence of the peripheral nerve involvement]. AB - A 41-year-old female of mitochondrial myopathy characterized by recurrent paralytic ileus and atonic bladder with the evidence of peripheral nerve involvement was described. This patient was admitted to our hospital because of the episode of paralytic ileus and atonic bladder at the age of 40 and 41 (1987). She had noticed sporadic headache from 1967, constipation from 1977, tinnitus and hearing disturbance from 1984. One month after her second admission in 1987, her symptoms of paralytic ileus and atonic bladder gradually disappeared. She was then transferred to the department of neurology for the evaluation of underlining neurological disorders. Neurological examination revealed dementia, oro-lingual dyskinesia, and proximal muscular weakness. However, none of the following signs or symptoms were observed; Ophthalmoplegia, blepharoptosis, retinitis pigmentosa, myoclonus, cerebellar ataxia, sensory disturbance, and orthostatic hypotension. Deep tendon reflexes were normal. Planter responses were flexor. Pyruvate and lactate were elevated in both serum and cerebrospinal fluid. Brain CT scan displayed moderate cerebral atrophy and basal ganglia calcifications. EMG was normal except for the external anal sphincter muscles which showed a denervation pattern. Motor nerve conduction velocity was normal in the right median and the right peroneal nerves. Sensory nerve conduction velocity was also normal in the right median and the right sural nerves. However, the amplitude of sensory potential was low in both these nerves. Atonic type of neurogenic bladder was noted on cystometry. There was a lack of voiding desire. The number of active sweat glands iontophoretically stimulated by pilocarpine was reduced. The most prominent feature of the muscle biopsy (the left biceps brachii) was myopathic changes with ragged-red fibers.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551556 TI - Uptake of Tc-99m MDP by adrenal metastases from undifferentiated large-cell carcinoma of the lung. PMID- 2551557 TI - Hepatoma: Tc-99m DISIDA uptake in primary and metastatic lesions. PMID- 2551558 TI - Tc-99m MDP uptake in a case of hepatoblastoma. PMID- 2551559 TI - A new method for isolation of human lymphocyte subsets reveals differential regulation of beta-adrenergic receptors by terbutaline treatment. AB - We adapted a technique for isolation of mononuclear leukocyte (MNL) subsets with immunomagnetic beads to the study of beta-adrenergic receptors. Mixed MNL cells were sequentially incubated with monoclonal antibodies specific for certain MNL subsets. Sheep antimouse antibodies coupled to magnetic beads were then added, and the desired MNL subset was pulled out with a magnet. This method yielded subsets with high purity and did not alter beta-receptor density or function. Healthy volunteers were treated for 7 days with the beta 2-selective agonist terbutaline (5 mg t.i.d.). Terbutaline treatment decreased beta-receptor number and isoproterenol-stimulated cyclic adenosine monophosphate (cAMP) generation in natural killer cells, helper T cells, and suppressor/cytotoxic T cells but not in B cells. The decrease was greatest in suppressor/cytotoxic T cells and least in helper T cells. Thus beta-adrenergic receptor regulation by agonists appears to differ among MNL subsets. PMID- 2551560 TI - Comparison of intravenous dilevalol with placebo in moderate hypertension. AB - Dilevalol, an agent that combines nonselective beta-blocking and beta 2-mediated vasodilating properties, was compared with placebo in 16 subjects with moderate hypertension in a double-blind crossover study. Dilevalol or a placebo was administered intravenously in bolus injections of 25, 50, and 50 mg at 15-minute intervals. Fifteen minutes after a cumulative dose of 125 mg, the blood pressure was lowered by 11/9 mm Hg, compared with 2/1 mm Hg after placebo (p less than 0.01 between groups for systolic and diastolic blood pressure), an effect that persisted for at least 105 minutes. Standing systolic blood pressure was also lowered in dilevalol-treated patients without orthostatic symptoms. No significant effects on heart rate were noted. Fifteen minutes after the last dose of dilevalol, plasma norepinephrine levels increased from a baseline of 200 +/- 24 to 495 +/- 44 pg/ml (p less than 0.01), compared with a nonsignificant rise from 262 +/- 21 to 306 +/- 28 pg/ml with placebo vehicle. Dilevalol also increased alpha-human atrial natriuretic factor by 5.4 pg/ml, compared with 0.5 pg/ml after placebo (p less than 0.01 between groups). Plasma renin activity and plasma epinephrine, aldosterone, and cyclic guanosine monophosphate levels were unchanged by dilevalol. There were no significant adverse effects with dilevalol administration. Compared with placebo, dilevalol given intravenously appears to be safe and effective antihypertensive treatment. PMID- 2551561 TI - Radiological appearances of cytomegalovirus infections. AB - The chest and abdominal radiographs of 35 patients with cytomegalovirus (CMV) infection were reviewed. The radiological appearances of cytomegalovirus pneumonia varied widely with no pattern sufficiently characteristic to allow early differentiation of CMV from other infective agents. Spontaneous pneumothorax and pneumomediastinum were seen frequently in association with advanced CMV pneumonia. Abdominal abnormalities included bowel wall thickening, dilated bowel loops and pneumatosis intestinalis. PMID- 2551562 TI - Lipiodol enhanced CT scanning in assessment of hepatocellular carcinoma. AB - We studied 20 patients with computed tomography (CT) following the intra-arterial injection of Lipiodol. Comparison with conventional CT, angiography and plain abdominal radiographs revealed additional satellite tumours in four patients. Technical difficulties occurred in selective hepatic arterial catheterisation and in satisfactory delivery of Lipiodol to all segments of the liver in a third of our patients. Plain abdominal radiographs following Lipiodol injection are inadequate in our experience in assessing tumour mass and distribution; we failed to demonstrate retention of Lipiodol by tumour tissue for longer than a few weeks in any case. The pattern of Lipiodol uptake within the tumour is not related to histology but to the vascular supply and size of the tumour. Our findings do not accord with the Japanese experience and this small study suggests that Lipiodol enhanced CT should be reserved for patients with small tumours in non-cirrhotic livers who are being considered for resection of their tumour. Such patients are likely to be encountered rarely in UK practice. PMID- 2551564 TI - [Rhinopharyngeal angiofibroma. Report of a case and review of the literature]. AB - The authors review the literature concerning rhinopharyngeal angiofibroma, discussing the etiology, diagnosis, medical and surgical management. They also report a case treated with anterior suprahyoid pharyngotomy according to Bocca. PMID- 2551563 TI - Lipiodol retention at the site of liver biopsy; a false positive result in the investigation of hepatocellular carcinoma. AB - A case is described in which Lipiodol, selectively injected into the hepatic artery as part of the investigation of a patient with hepatocellular carcinoma, was retained at the site of a previous liver biopsy leading to the false positive diagnosis of a daughter nodule. This has not been previously described. PMID- 2551566 TI - Genital herpes infections. AB - Genital herpes infections can be diagnosed by clinical criteria, but in most cases laboratory methods are superior. This article reviews the current methods available for detection of herpes simplex virus and for documenting infection by serology. The applicability and limitations of these methods are discussed. PMID- 2551565 TI - [Comparison of the efficacy of captopril and enalapril in single doses in the treatment of arterial hypertension. Evaluation by means of non-invasive ambulatory monitoring]. AB - The study was designed to compare the efficacy of captopril and enalapril, both orally active inhibitors of angiotensin converting enzyme, in the treatment of primary hypertension when administered in a single daily dose. After placebo washout for two weeks, 20 hypertensive patients (I-II class, according to WHO), were admitted to active treatment, in a randomized sequence, with captopril (50 mg) and enalapril (20 mg) once a day in the morning (8 a.m.). Supine and erect blood pressure and heart rate were measured weekly, 24 hours after drug administration by using a mercury standard sphygmomanometer. In all patients ambulatory noninvasive blood pressure monitoring was performed after 4 weeks of treatment. The data confirmed the efficacy of both drugs in lowering blood pressure. However, while the antihypertensive effect of enalapril was prolonged throughout 24 hours, captopril was effective only for about 22 hours, a period longer than previously suggested on the basis of serum ACE inhibition, but not sufficient to cover the whole day. Therefore, if captopril therapy has to be used in a single daily dose an attempt should be made using an increased dosage or by employing the drug in some retarded pharmaceutical form. The need to prolong the antihypertensive effect of captopril to 24 hours is based on the clinical experience according to which the smaller the number of tablets to be taken the better the compliance. This is particularly true for cases of asymptomatic hypertension which nevertheless require lifelong therapy. PMID- 2551567 TI - Genital human papillomavirus infections. AB - Genital human papillomavirus (HPV) infection is an increasingly common sexually transmitted disease that has been associated with carcinoma of the cervix. Genital warts are easily diagnosed by clinical criteria; however, the more prevalent subclinical infections generally require laboratory diagnosis. Satisfactory methods for culture of HPV do not exist, and cytologic and histopathologic methods are the most widely available diagnostic techniques. Detection of HPV DNA is a more sensitive method, but its role in the clinical laboratory remains to be defined. PMID- 2551568 TI - Cytopathology of sexually transmitted disease. AB - Although other methods exist for routine identification of most sexually transmitted diseases, the pathologist can often recognize the etiologic agents or their cytologic effects on Papanicolaou (Pap) smear. Useful clinical information requires both the ability to recognize these effects and knowledge of their limitations. Pap smear changes characteristic of Trichomonas vaginalis, Chlamydia trachomatis, herpes simplex virus, Gardnerella vaginalis, Candida sp., and human papillomavirus infections are described and their diagnostic accuracy evaluated. PMID- 2551569 TI - Evaluation of a new herpes simplex virus typing reagent for tissue culture confirmation. AB - Monoclonal antibody typing reagents for herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) from California Integrated Diagnostics, Inc. (Berkeley, CA) were compared to two other commercially available HSV monoclonal antibody typing reagents. Of the 105 specimens tested, of which 81 were positive for HSV, there was 100% agreement with all three typing reagents. PMID- 2551570 TI - Correlation of urine pH with the detection of cytomegalovirus by the shell vial technique. AB - The influence of the pH of urine on the detection of cytomegalovirus (CMV) by the shell vial assay was evaluated. The pH values of 295 urine specimens ranged from 4.7 to 8.5 (mean 6.3) and were not significantly different in culture-positive versus culture-negative samples. None of the urine specimens appeared to be toxic for the cells used in the shell vial assay. We recommend inoculation of urine specimens into shell vials without adjustment of pH. PMID- 2551571 TI - Several methods of detecting varicella-zoster virus (VZV) and measuring VZV immune status. PMID- 2551572 TI - Synthesis of latent collagenase and collagenase inhibitor by bovine aortic medial explants and cultured medial smooth muscle cells. AB - Bovine aortic medial tissue and medial smooth muscle cells were demonstrated for the first time to synthesize a latent collagenase together with collagenase inhibitor in culture. Molecular weights of the latent collagenase and its inhibitor derived from aortic medial tissue explant were estimated to be about 52 K by gel filtration and 26.5 K by electrophoresis, respectively. Activated aortic collagenases cleaved type I collagen in solution into 3/4 (alpha A) and 1/4 (alpha B) length cleavage fragments and were inhibited by EDTA, o-phenanthroline, dithiothreitol, bovine serum, and highly purified dental pulp and aortic collagenase inhibitors. The aortic inhibitors showed inhibitory activity against all the animal collagenases tested, except for bacterial collagenase. Double immunodiffusion analysis using a monospecific antiserum prepared against dental pulp inhibitor showed that the aortic inhibitors are immunologically identical to the pulp inhibitor. Using the same antiserum, we found immunoreactive collagenase inhibitor protein to be localized along the collagen fibers between elastic membranes in aortic medial tissue. PMID- 2551573 TI - Isolation of a herpes simplex virus type 2 that is retinovirulent in mice. AB - The virulence of a herpes simplex virus type 2 (HSV-2) isolated from the urine of a patient (SL) with acquired immunodeficiency syndrome (AIDS) and bilateral acute retinal necrosis (ARN), was investigated in mice. The ratio of plaque forming units (PFU) in fibroblasts to the 50% lethal dose (LD50) of HSV-2(SL) in mice was 10 fold more than the PFU to LD50 ratio of a neurovirulent HSV-2, strain 186. Further, HSV-2(SL) caused retinitis with and without lethal encephalitis in mice inoculated intracranially (i.c.). In contrast, mice inoculated with HSV-2(186) died of encephalitis without ocular disease. HSV-2(SL) was isolated from eye and/or brain tissue 1 to 15 days post i.c. inoculation. Ocular disease progressed from an initial mild chorioretinitis on day 8 to total retinal necrosis with panuveitis by day 11 in mice given 10 PFU of HSV-2(SL) i.c. HSV antigen was detected initially in the cells of the optic nerve and spread into the ganglial cells of the nerve fiber layer, the neurosensory cells of the inner nuclear layer, and the cells of the retinal pigment epithelium (RPE) between days 8 and 10. Thus, this study supports the concept that HSV neurovirulence varies between strains and presents a HSV-2 neurotransmission animal model of ARN. PMID- 2551574 TI - (Na+ + K+)-ATPase and opsin in retinas of RCS dystrophic rats: time course study. AB - Retinas of Royal College of Surgeons (RCS) dystrophic rats were investigated immunocytochemically for the distribution of the membrane-bound enzyme (Na+ + K+) ATPase and the photo-pigment opsin prior to and during the retinal disease process. Retinas of 11 day-old dystrophic and control Long Evans rats showed (Na+ + K+)-ATPase immunostain most dense in the outer nuclear layer (ONL) and inner plexiform layer (IPL). Also, in these retinas, immunostaining for opsin was dense along rod inner segments (RIS) and on plasmalemma of ONL cell bodies and several cell bodies in the inner nuclear layer (INL). In retinas of control rats at 30 days and later, less dense (Na+ + K+)-ATPase immunostain was detected in the ONL than at 11 days and opsin-immunostained ROS were also detected. However, (Na+ + K+)-ATPase immunostained RIS were shorter in retinas of 30 day-old dystrophic than in retinas of age-matched control rats and an opsin-immunostained debris zone was also observed in dystrophic retinas. In retinas of 60 day-old dystrophic rats, the opsin-immunostained debris zone was more prominent than at 30 days, while the few ONL cell bodies immunostained for both proteins. Also, the outer IPL of 60 day-old dystrophic rat retinas immunostained more densely for (Na+ + K+)-ATPase than the inner IPL. In 120 day-old dystrophic rat retinas, (Na+ + K+) ATPase immunostain was detected in the INL and IPL, while opsin staining was demonstrated only in the debris zone. This opsin-immunostained debris disappeared in a central-to-peripheral gradient. (Na+ + K+)-ATPase immunostain was still present in the INL and IPL in retinas and the optic nerve of one year-old RCS dystrophic rats; however, opsin was restricted to a few surviving cell bodies in the peripheral retina. PMID- 2551575 TI - Cytomegalovirus replication in cultured human retinal pigment epithelial cells. AB - Retinal pigment epithelium (RPE) was isolated from the globe of a donor positive for human immunodeficiency virus (HIV) who had cytomegalovirus (CMV) retinitis secondary to acquired immunodeficiency syndrome (AIDS). In culture, the cells exhibited normal epithelioid morphology by phase contrast microscopy. After two weeks the cells developed cytomegaly and dense intranuclear and cytoplasmic inclusions and, eventually, died. Transmission electron microscopy (EM) demonstrated intranuclear nonenveloped virus particles 80-120 nm in diameter consistent with a herpes type infection. Immunofluorescence staining demonstrated the presence of CMV antigens. Conditioned medium from the infected cells caused infection in RPE cells isolated from normal donors. Hybridization assay demonstrated the presence of CMV DNA and indicated that the time course of the infection was similar, but not identical to infection in MRC-5 and HEL cells. We conclude that cultured human RPE is a permissive host for CMV. PMID- 2551576 TI - The relationship of corneal Langerhans cells to herpes simplex antigens during dendritic keratitis. AB - The corneal migration and topographic distribution of Langerhans cells (LC) in relation to herpes simplex virus antigens was studied during the course of dendritic keratitis in inbred mice. Corneal epithelial sheets from infected mice at selected time points were "double stained" for Ia-positive Langerhans cells and HSV antigens, using a sequential avidin biotin immunoperoxidase and glucose oxidase technique. The amount of HSV antigen was maximum at day 2 paralleling the clinical time course, with most corneal epithelium HSV antigen negative by day 8. LC were seen in peripheral corneas by day 2 and in paracentral and central cornea by day 8, with peak numbers detected between days 8 and 11 post-infection. Although HSV antigens and LC were simultaneously detected within corneal epithelium, LC were not observed in anatomic juxtaposition to HSV antigens, even after reinoculation of infected corneas with HSV on day 14 following the primary infection. These data suggest that local factors in the corneal epithelium other than HSV antigens per se may be chemotactic for LC during the course of dendritic keratitis. PMID- 2551577 TI - Testicular cancer. AB - Although testicular cancer is a relatively rare tumor, it is the most common cancer among men aged 15 to 35 years. In the United States in 1989 approximately 5,500 men will be diagnosed with testicular cancer. Up until 2 decades ago, testicular cancer was the most common cause of a cancer death in this age group. The advances in diagnosis and treatment that will be described in this monograph represent one of the major achievements in the treatment of solid tumors. Testicular cancer is now one of the most curable of all cancers; the 5-year relative survival rate is in excess of 90%. In the U.S. fewer than 500 men will die from this disease in 1989. The primary goals in the treatment of testicular cancer as the 1990s approach will be to further decrease the mortality from this disease and to decrease the morbidity of treatment that has led to this dramatic improvement in survival. PMID- 2551578 TI - The mutagenic and immortalizing potential of polyoma virus large T antigen. PMID- 2551579 TI - Structure, activity, and regulation of the bovine papillomavirus E5 gene and its transforming protein product. PMID- 2551580 TI - Functional and sequence similarities between HPV16 E7 and adenovirus E1A. PMID- 2551581 TI - Two independently transforming functions of human papillomavirus 8. PMID- 2551582 TI - Human papillomavirus early gene products and maintenance of the transformed state of cervical cancer cells in vitro. PMID- 2551583 TI - A genetic analysis of the zinc finger of SV40 large T antigen. PMID- 2551584 TI - Epstein-Barr virus DNA replication. PMID- 2551585 TI - Characterization of the BNLF-1 oncogene of Epstein-Barr virus. PMID- 2551586 TI - Genes for the synthesis of deoxythymidylate monophosphate in T-cell lymphoma inducing herpesviruses of nonhuman primates. PMID- 2551587 TI - Analysis of hepatitis B virus gene functions in tissue culture and in vivo. PMID- 2551588 TI - Transactivation by hepatitis B virus may contribute to hepatocarcinogenesis. PMID- 2551589 TI - Effect of amino acid phosphorylation on the DNA binding properties of large T antigen. PMID- 2551590 TI - SV40 T antigen catalyzed duplex DNA unwinding. PMID- 2551591 TI - Control of transcription in vitro from simian virus 40 promoters by proteins from viral minichromosomes. PMID- 2551592 TI - Dissection of the T antigen/mouse p53 complex and its inhibitory effects on viral origin-directed DNA replication in vivo and in vitro. PMID- 2551593 TI - Interactions between SV40 and cellular oncogenes in the transformation of primary rat cells. PMID- 2551594 TI - Metabolic stabilization of p53 in SV40-transformed cells correlates with expression of the transformed phenotype but is independent from complex formation with SV40 large T antigen. PMID- 2551595 TI - SV40 large T antigen induces a protein kinase responsible for phosphorylation of the cellular protein p53. PMID- 2551596 TI - Evidence for threshold effects in transformation of pancreatic beta cells by SV40 T antigen in transgenic mice. PMID- 2551597 TI - Structure and function of SV 40 large T antigen: communication between functional domains. PMID- 2551598 TI - Diffuse pulmonary infiltrates following lymphography. PMID- 2551599 TI - Face-mask CPAP and sodium bicarbonate infusion in acute, severe asthma and metabolic acidosis. AB - A patient with a history of previous mechanical ventilation for asthma was hospitalized with acute, severe asthma; metabolic acidosis; and hyperlactatemia. In addition to standard therapy, she was successfully treated with face-mask CPAP and IV sodium bicarbonate infusion. This therapy may help prevent mechanical ventilation in some patients with acute, severe asthma and lactic acidosis. PMID- 2551600 TI - Newer agents against methicillin and/or gentamicin-resistant and -susceptible staphylococci. AB - The in vitro activities of the newer agents Deptomycin, Difloxacin, A-56620 and Ciprofloxacin were compared with those of vancomycin, rifampin and gentamicin against methicillin-susceptible and -resistant Staphylococcus aureus and coagulase-negative staphylococci. All isolates were susceptible to vancomycin, rifampin, deptomycin and the quinolones tested. Newer agents had activity superior to that of vancomycin, whereas the activity of the newer agents and rifampin was comparable. Gentamicin resistance was frequent, however, the resistant staphylococci were susceptible to other antibiotics tested. PMID- 2551601 TI - Computed tomography of parotid masses. AB - Fifty-six patients with parotid masses, benign tumors in 22, malignant tumors 26 and other diseases 8, were examined by CT scan, which can differentiate intrinsic from extrinsic parotid lesions, especially deep lobe from parapharyngeal tumors and typical benign tumors from malignant ones, but may fail to differentiate aggressive benign tumors from low-grade malignant ones. CT sialography was done to detect the relation between the trunk of the facial nerves and tumor according to the band of normal parotid tissue between the tumor and posterior border of the mandibular ramus, and incremental dynamic CT with intravenous bolus injection of contrast medium to detect the relation between the tumor and carotid sheath. We conclude that the images of CT sialography are much better than those of plain CT and intravenous enhancement CT. PMID- 2551603 TI - Serological analysis and biochemical characterization of monoclonal antibodies defining antigens of human hepatocellular carcinoma. AB - A panel of seven murine monoclonal antibodies reactive with human hepatocellular carcinoma (HCC) cell line, SK- HEP-1, resulted in the definition of four distinct antigen systems, designated HB4, HB5, HB1 and HJ2. HB4 antigen was found to be expressed specifically on HCC cell lines and fresh HCC specimens but not on normal liver. Immunoprecipitation tests suggest that the HB4 epitope may be a heat-stable carbohydrate determinant on a high molecular mass molecule. HB5 antigen was found to have less-restricted expression on a panel of normal adult tissues and on melanoma, astrocytoma, sarcoma, neuroblastoma and epithelial cancer cell lines. In fetal and adult liver, HB5 antigen localized to bile canaliculi and ducts. Under reducing conditions, three mAbs detected a Mr 140,000 glycoprotein using lysates of [125-I], [3-H]-glucosamine and [35-S]-methionine labeled SK-HEP-1 cells. Under non-reducing conditions an additional component of greater than Mr 200,000 was also detected. HB1 antigen was found on almost all monolayer cell lines and not on most cultured suspension cells. This antigen was also detected on cultured HCC cells inoculated into nu/nu mice. Immunoprecipitation experiments revealed that the HB1 antigen is a bimolecular complex with an Mr 170,000 alpha chain and Mr 130,000 beta chain under non reducing conditions, and three subunits of Mr 140,000, Mr 30,000 and Mr 130,000 under reducing conditions. Two antibodies reacted with epitopes on the alpha chain. HJ2 antigenic determinant is a heat-stable component which could not be immunoprecipitated. This most widely expressed antigen was found in secreted form in many of the cells and tissues examined. These antibodies introduce new antigens which may serve as useful markers for the diagnosis, classification and investigation of HCC and other liver diseases. PMID- 2551602 TI - Pre- and post-operative hypothalamic-pituitary-thyroidal axis function in patients with prolactinoma, growth hormone tumour and ACTH tumour. AB - Pre- and postoperative hypothalamic-pituitary-thyroid axis function was studied in 38 patients with pituitary adenomas (PRL, GH and ACTH tumours), of whom 35 were surgically confirmed and three diagnosed by clinical signs, CT scanning and hormone assessments. About ten days after operation, the same study was repeated in 10 patients with prolactinoma and 7 with growth hormone (GH) tumour. The preoperative abnormal serum TSH response to TRH was found in 8/20 patients with prolactinoma, 9/16 with GH tumour, and 2/2 with Cushing's disease due to ACTH microadenoma. The incidence of abnormal TSH response to TRH was not significantly increased in patients with larger adenoma in either PRL or GH tumour group. In 8 cases of prolactinoma, metoclopramide (MCP, 10 mg, P.O.) test was also performed and there was a significant positive correlation between TSH responses to TRH and to MCP. Serum TT3 in the GH tumour group was within normal ranges, but significantly higher than that of the normal and prolactinoma groups. After operation, TT3 was significantly decreased as compared with that before operation and there were marked changes in TSH response to TRH. In conclusion, there were some abnormalities in TSH control in patients with non-TSH pituitary tumour, and in serum TT3 control in patients with GH tumour. The surgical treatment of pituitary adenoma can lead to transient decrease in TSH reserve and serum TT3 level probably resulting from both stress and/or destruction of thyro-trophs by the operation. PMID- 2551604 TI - [Prognostically relevant factors in ductal pancreatic cancer]. AB - Prognosis of patients with pancreatic carcinoma is bad. Less than 1% of all patients and even after pancreas resection only 10% survive more than 5 years. To evaluate the factors which influence the prognosis we analyzed retrospectively all patients who were operated on a pancreas cancer (n = 484) from January 1, 1978 to December 31, 1987. Age, sex, perineural invasion and lymphangiosis carcinomatosa had no prognostic influence. The duration of symptoms, degree of cellular differentiation, tumor size, preoperative level of the tumor associated antigen CA-19-9, nodal involvement and stage were statistically significant for prognosis. R1 and R2-resections had no better prognosis than other palliative procedures. PMID- 2551605 TI - [Resection of the superior vena cava in tumor occlusion]. AB - Report about three cases of resection of the superior vena cava for tumor stenosis or occlusion. In one patient no reconstruction was performed while in the other two interposition of an armored PTFE-prosthesis between right brachio cephalic vein and vena cava superior was carried out. In agreement with other authors it is concluded that best results can be obtained by reconstruction with a PTFE-prosthesis or autologous vein graft. PMID- 2551606 TI - Role of carbohydrates in receptor-mediated fertilization in mammals. AB - The mouse sperm receptor, called ZP3, is a glycoprotein (83,000 Mr) that consists of a 44,000 Mr polypeptide chain (402 amino acids), three or four N-linked oligosaccharides, and an undetermined number of O-linked oligosaccharides. There are more than 10(9) copies of ZP3 present throughout the mouse egg extracellular coat, or zona pellucida. As a prelude to fertilization, each acrosome-intact sperm binds in a relatively species-specific manner to tens-of-thousands of copies of ZP3 at the surface of the zona pellucida. Binding to ZP3 induces sperm to undergo the acrosome reaction (membrane fusion) and, consequently, enables them to penetrate through the zona pellucida and to reach, and then fuse with, egg plasma membrane (fertilization). Purified ZP3, as well as a specific class of ZP3-derived O-linked oligosaccharides (3900 Mr), exhibit sperm receptor activity in vitro. The oligosaccharides, which represent a relatively low percentage of total ZP3 O-linked oligosaccharides, account for the glycoprotein's sperm receptor activity in vitro (i.e., recognition and binding). Furthermore, either enzymic removal or modification of certain sugars that constitute these oligosaccharides results in destruction of sperm receptor activity. These and other findings strongly suggest that during mammalian fertilization carbohydrates play a fundamental role in species-specific sperm-egg interactions. PMID- 2551607 TI - Multifunctional glycoprotein receptors for insulin and the insulin-like growth factors. AB - Insulin and the insulin-like growth factors (IGF) I and II are structurally related peptides that elicit a large number of similar biological effects in target cells. Three well-characterized receptor complexes bind one or more of these peptides with high affinity. Two of these receptors, denoted as type I, are ligand-activated tyrosine kinases with similar heterotetrameric alpha 2 beta 2 subunit structures which bind insulin or IGF-I, respectively, with highest affinity. Ligand-stimulated tyrosine autophosphorylation of these receptors further activates their intrinsic tyrosine kinase activities both in vitro and in intact cells. Rapid signal transduction follows such receptor autophosphorylation and tyrosine kinase activation, leading to increased serine phosphorylation of many cellular proteins and decreased serine phosphorylation of several others. Experiments in our laboratory have identified three distinct insulin-activated serine kinase activities in cell-free extracts that appear to account for the insulin-stimulated serine phosphorylation of the insulin receptor itself, ATP citrate lyase, and acetyl CoA carboxylase, respectively. A third receptor in this group binds IGF-I and II, lacks kinase activity and is denoted as type II IGF receptor. Amino acid sequences of this receptor deduced from isolated rat cDNA clones show a high degree of homology with those of the bovine cation-independent mannose 6-phosphate (Man-6-P) receptor. We demonstrated that these receptors are indeed identical. The IGF-II/Man-6-P receptor rapidly recycles between the cell surface membrane and intracellular membrane compartments, providing for the rapid uptake of both IGF-II and mannose 6-phosphate-linked lysosomal enzymes. Insulin action markedly increases the proportion of receptors in the plasma membrane and the uptake of bound ligands. We also observe that large amounts of the extracellular domain of the IGF-II/Man-6-P receptor are released into the serum of fetal, neonatal and adult rats. The biological role of this receptor in IGF-II function is yet to be determined. PMID- 2551608 TI - Multiple subfamilies of carbohydrate recognition domains in animal lectins. AB - Calcium ion-dependent carbohydrate recognition domains (CRDs) are found in a range of proteins including receptors for serum glycoproteins and proteoglycans of the extracellular matrix. These C-type CRDs have homologous amino acid sequences characterized by the presence of certain invariant residues. Analysis of the genes for five of the proteins reveals that in each case the CRD-coding sequence is separated from the rest of the gene by an intron. The genes fall into two groups: those in which the coding sequence for the CRD is interrupted by two introns, and those in which the coding sequence is contained in a single exon. The sequences of domains in each category are consistent with the suggestion that the different gene structures reflect early evolutionary divergence of two subfamilies of C-type CRDs in animal lectins. However, carbohydrate-binding specificity does not directly parallel the evolutionary categorization. Comparison of the primary structures of CRDs in each subfamily which have related binding specificities may help to identify residues involved in ligating carbohydrates. This type of analysis is being extended by the use of bacterial expression systems to investigate in greater detail the binding characteristics of the CRDs. PMID- 2551609 TI - Glycoprotein oligosaccharides as recognition structures. AB - A series of observations--the pronounced changes in the expression and distribution of oligosaccharide antigens during embryonic development, cell differentiation and oncogenesis, the prominence of these changing structures (oncodevelopmental antigens) on the receptor for epidermal growth factor, and the stimulation of receptor autophosphorylation following their perturbation with antibodies--has suggested that the oligosaccharides of growth factor receptors and complementary lectins may be intimately involved in molecular recognition events in growth and differentiation processes. For elucidating oligosaccharide recognition by diverse cellular and secreted proteins and microbial adhesins, a new technique has been developed which involves the overlay of immobilized oligosaccharide probes (neoglycolipids) derived from glycoproteins and other sources. New insights have been gained into carbohydrate recognition by several mammalian lectins, and a novel receptor system has been discovered in Escherichia coli isolated from patients with urinary tract infections. This new technique seems ideal for elucidating oligosaccharide recognition in diverse biological settings, and for 'quality control' of the sugar chains of recombinant glycoproteins engineered for the purpose of administration to man. PMID- 2551610 TI - Binding modes of mammalian hepatic Gal/GalNAc receptors. AB - Mammalian Gal/GalNAc receptors show dramatic preference for three-branched oligosaccharide structures over two- or one-branched counterparts. The spatial arrangement of the Gal residues is extremely important for optimal binding. The three terminal Gal residues in the best triantennary ligand are about 15-30 A apart, and therefore the sugar-combining sites on the receptor may also have the same geometry. The results obtained with synthetic ligands containing Gal or GalNAc are in agreement with this concept. Photoaffinity labelling and GalNAc lactoperoxidase catalysed iodolabelling of the receptors revealed that the minor subunits (52 and 60 kDa) were more readily labelled than the major subunit (43 kDa). The stoichiometry of binding was determined with synthetic ligands containing GalNAc. The results indicated that each subunit may have two sugar combining sites. A model for subunit assembly is proposed on the basis of these results and the finding that coexpression of all subunits is necessary for the binding and processing of asialo-orosomucoid in transfected cells, whereas Gal polylysine can be bound and processed by the cells expressing only the major subunit. PMID- 2551611 TI - [Effect of cyclic nucleotides (cAMP) on experimental HSV-I keratitis]. AB - The authors report an experimental study on the effect of cAMP on HSV-I keratitis. Rabbit corneas developed typical dendritic keratitis on the 3rd day after inoculation of HSV-I. On the 9th day, the cAMP level in plasma significantly decreased from 174.9 +/- 20.2 to 52.1 +/- 18.2 pmol/ml, while the cGMP level increased. The cAMP level in aqueous humour also decreased, with an increase in cGMP level increased. The cAMP level in aqueous humour also decreased, with an increase in cGMP level. cAMP was used subconjunctivally for treatment of HSV-I keratitis. 48 hours after injection, the cAMP level in aqueous humour markedly rose from 11.1 +/- 2.0 to 35.6 +/- 12.9 pmol/ml and 6 days later to 65.0 +/- 30.9 pmol/ml, with concurrent decrease in cGMP level, and the ratio of cAMP/cGMP increased from 1.6 +/- 0.6 to 15.3 +/- 4.6. In the group of treatment by acyclovir, the level of cAMP in aqueous humour increased by 9.5 times after 3 days of treatment, with no remarkable effect on the cGMP level. These results showed that cAMP was equally effective as ACV in treatment of experimental HSV 1 keratitis. The antiviral action of cAMP was discussed and the pharmacological effect of cAMP approached. PMID- 2551612 TI - Effect of OK-432 on large-bowel carcinogenesis in rats. AB - To examine the carcinogenetic and growth inhibitory effects of OK-432, large bowel carcinoma was induced experimentally in rats by intrarectal injection of N methyl-N-nitrosourea (MNU), and OK-432 was administered intradermally. Rats were sacrificed after six months and the large intestine was cut into serial sections. Histopathologic examination and analysis of the infiltrating mononuclear cells, using monoclonal antibodies, were performed. The average rate of carcinogenesis per rat was 15.7 +/- 8.5 in the MNU alone group (n = 10) and 8.3 +/- 3.5 in the MNU and OK-432 group (n = 6). The invasion was deeper than the muscularis propria in 16 out of 157 lesions (10.2 percent) in the MNU alone group and in one out of 50 lesions in the MNU + OK-432 group (2.0 percent) (P less than 0.05). When time of appearance of atypical glands or carcinomas were compared in the MNU alone and MNU + OK-432 group, carcinogenesis was found to be delayed in the MNU + OK-432 group. In the investigation of infiltrating mononuclear cells using monoclonal antibodies, there were increases in helper T cells in both the MNU alone and MNU + OK-432 groups, but there was little difference between the two groups. The results of this study suggest that the suppression of experimental carcinogenesis in the large bowel by the concomitant administration of OK-432 with MNU, may be due to the enhanced activation or prolonged activated state of immunocompetent cells, which appear via antigen recognition, by OK-432. PMID- 2551613 TI - Familial adenomatous polyposis in association with thyroiditis. Report of two cases. AB - The authors report two patients with familial adenomatous polyposis and thyroiditis. One patient was discovered at autopsy to have in addition, a follicular carcinoma of the thyroid and focal nodular hyperplasia of the liver. The other patient had a sister with familial adenomatous polyposis and a papillary carcinoma of the thyroid. The association between familial adenomatous polyposis and thyroiditis has not been previously reported. PMID- 2551615 TI - Proinsulin levels and insulinomas. PMID- 2551614 TI - Reappraisal of surgical treatment of small hepatocellular carcinomas in cirrhosis: clinicopathological study of resection or transplantation. AB - Thirty-two patients with hepatocellular carcinoma (HCC) occurring in individuals with cirrhosis had a potentially curative surgical procedure. Twenty-two had segmental hepatic resections (HR), and 10 underwent orthotopic liver transplantation (OLTx). The diagnosis of hepatic malignancy was established in each case preoperatively, and each case was studied intraoperatively by means of sonography. Postoperatively each surgical specimen was examined pathologically with attention to the possibility of intrahepatic tumor spread. Twenty-three of the 32 patients had single small HCC lesion (less than 5 cm diameter) identified preoperatively. Sixteen of these underwent HR and seven underwent OLTx. Multiple additional neoplastic lesions were found in 19% of the 16 HR cases and in 14% of those undergoing OLTx when the resection specimens were examined pathologically. Vascular invasion was present in 43% of the OLTx patients and in 25% of the HR patients. Subtotal hepatic resection for small HCC occurring in cirrhosis has produced few long-term survivals. Both pre- and intraoperative sonography have been shown to underestimate the extent and distribution of these tumors. Based upon this experience that (1) vascular spread occurs often in HCC and (2) a high risk of postoperative hepatic failure can be expected after HR in cirrhotic individuals, OLTx is the most rational surgical procedure for such cases as it has the potential to cure. PMID- 2551616 TI - Aspiration cytology of neuroendocrine tumors below the diaphragm. AB - In a series of 630 transabdominal fine-needle aspirates, 24 neuroendocrine tumors (NETs) presenting below the diaphragm were encountered. Of these, 10 were classic small-cell undifferentiated carcinomas (SCUCs). Recognition of SCUC in abdominal aspirates has major directive value, pointing to bronchogenic origin, possibly unsuspected, in most cases (eight of 10 in this series). The other 14 NETs were 10 pancreatic islet-cell tumors and four intestinal carcinoids, both diagnosed more frequently in metastatic than in primary sites (10 of 14 in the liver in this series). The cytologic presentation of these non-SCUC neoplasms has received little literature attention, and it is described and illustrated here, with discussion of cytologic pitfalls and differential diagnoses. Diagnostic specificity was 100%, and sensitivity was also 100%, exclusive of two sampling errors. The purpose of this article is to establish a broader view of the range of morphological features of the extended neuroendocrine group of tumors in aspiration cytology. PMID- 2551617 TI - Aspiration cytology of Wilms' tumor: correlation of cytologic and histologic features. AB - We examined the cytomorphologic features of fine-needle aspiration biopsy (FNAB) specimens from 23 Wilms' tumor patients. The findings were correlated with histopathologic patterns from these tumors. The study revealed a close resemblance between the cytologic and histopathologic appearance of various cellular elements in Wilms' tumors. The major cellular patterns seen in Wilms' tumor include blastemal cells, blastemal cells with epithelial differentiation, blastemal cells with tubular differentiation, and stromal elements. It is hoped that recognition of these cellular components in aspiration smears will be helpful in establishing an FNAB diagnosis of Wilms' tumor. PMID- 2551618 TI - Human papovavirus in a routine urine specimen of a four-year-old boy. AB - Cellular changes produced by viruses can be readily identified using light microscopy and Papanicolaou stain of a fixed specimen. These findings can then be confirmed by viral culture and/or electron microscopy studies. Human polyomavirus, common in transplant recipients or otherwise immunocompromised patients, is one virus that can be identified using these methods. The following is a case study of a 4-yr-old boy with no known immune impairment who exhibited human papovavirus (polyomavirus) on a routine urine examination. The diagnosis was confirmed by electron microscopy. PMID- 2551619 TI - What are the various methods for HPV detection? PMID- 2551620 TI - Detection of human papillomavirus antigen on urinary sediments by means of immunoperoxidase staining. PMID- 2551621 TI - [A comparative analysis of the effect of the synthetic SKD, vasopressin and asparaginic acid on the electrically sensitive sodium and potassium ion channels of neuronal somatic membrane]. PMID- 2551623 TI - [The significance of the kallikrein-kinin system in the pathogenesis of essential hypertension]. AB - The levels of plasma kininase II (ACE), a high molecular weight-kininogen (HMW), alpha 2-mega-globular protein (alpha 2M), prekallikrein (PK), alpha 1-antitrypsin (alpha 1 AT) were determined in essential hypertension and control groups. The results showed that there is significant increase of ACE and PK activity and HMW content in patients with essential hypertension than in control group (P less than 0.001). No changes in alpha 2M and alpha 1AT were found. These results suggest that human plasma kallikrein system was inhibited in patients with essential hypertension. PMID- 2551622 TI - Changes in lymphocyte beta-adrenoceptor density after dilevalol oral treatment. AB - Dilevalol (R,R'-isomer of labetalol) is an antihypertensive agent combining non specific beta blockade with peripheral vasodilatation due to beta 2-receptor agonism. The aim of this study was to determine the effects of chronic dilevalol administration on lymphocyte beta 2-adrenoceptor density. The investigation was conducted as a double-blind, placebo-controlled comparison. Ten days of chronic dilevalol (400 mg) or placebo treatment was administered to 12 healthy normotensive volunteers. Clinical and biochemical parameters: heart rate (HR) systolic and diastolic blood pressure (SBP, DBP), electrocardiogram, norepinephrine (NE), epinephrine (E), MHPG (3-methoxy-4-hydroxyphenylethylene glycol; one of the major brain metabolites of NE) were analysed on the first day (D1) before (H0) and 3 h after oral treatment (H3) and on the tenth day (D10). Clinical results showed no significant changes in HR, DBP, SBP in the two groups (placebo, n = 6; dilevalol, n = 6). NE increased 3 h after the first oral dilevalol intake (p less than 0.05). This increase is greater than that due to the circadian variation observed in the placebo group. Acute dilevalol treatment seems to increase the plasma circulating NE level, which returns to normal values after 10 days of chronic treatment. Binding assays were performed before and after 10 days of treatment. In the placebo group, no change in beta-adrenoceptor density was observed (36.6 +/- 8.3 versus 38.3 +/- 9.4 femtomol/mg of protein). Lymphocyte beta-adrenoceptor density (Bmax) significantly decreased after 10 days of dilevalol treatment without any changes in affinity (KD). Results were 40.3 +/ 11.6 (D1) and 30 +/- 7.6 (D10) (p less than 0.05). It was concluded that dilevalol down-regulated lymphocyte beta 2-adrenoceptor density, suggesting that beta 2 agonism of dilevalol is predominant. PMID- 2551624 TI - [Liver calcifications in metastasizing vipoma]. AB - A 53-year-old woman had suffered from watery diarrhoea and substitution-requiring hypokalaemia for 18 months. Ultrasonography and abdominal x-ray revealed multiple calcifications in the liver. Retrograde cholangiopancreaticography, performed because an endocrine-secreting pancreatic tumour was suspected, demonstrated occlusion of the major pancreatic duct and calcification in the body of the pancreas. Gastrointestinal hormone levels were elevated (vasoactive intestinal polypeptide 950 pg/ml, pancreatic polypeptide 1000 pmol/l and neurotensin 86 pmol/l) in the blood, and immunohistochemical results on cells of an adenocarcinoma obtained by needle biopsy confirmed a metastasizing VIPoma. Administration of the somatostatin analogue SMS 201-995 achieved marked improvement. PMID- 2551626 TI - Integrins and cell adhesion molecules: neuronal receptors that regulate axon growth on extracellular matrices and cell surfaces. PMID- 2551625 TI - Receptor-mediated active adhesion to the substratum is required for neurite outgrowth. PMID- 2551627 TI - [Normalization of the phenotype of transformed Djungarian hamster cells during selection for colchicine resistance]. AB - The range of the Djungarian hamster cell lines selected for colchicine resistance in high doses (from 7 to 200 micrograms/ml) was studied. These cell lines are characterized by the different levels of drug-resistance (1000- to 16000-fold). A positive correlation is found between the reversion rate of malignant phenotype and the cellular drug-resistance level. Tumorigenicity and anchorage independence decrease with an increase of the drug-resistance level. The actin-containing bundles of microfilaments in more resistant cells are more organized. The 22 kD protein content increases with the drug-resistance level. The mechanisms of malignancy reversion are discussed. PMID- 2551628 TI - Characterization of tumor necrosis factor-alpha receptors in human and rat thyroid cells and regulation of the receptors by thyrotropin. AB - Administration of recombinant human tumor necrosis factor-alpha (TNF) to rats and mice produces a model of nonthyroid illness in which there is impairment of hypothalamic-pituitary thyroid function, including reduced serum concentrations of T4 and T3, reduced thyroid radioiodine uptake, and reduced response to TSH. In this study, we tested the binding and effects of TNF on FRTL-5 cells and on four human thyroid carcinoma cell lines. The TSH-stimulated [125I]iodide uptake by FRTL-5 cells was inhibited by TNF in a dose-dependent manner. The four human thyroid carcinoma cell lines (NPA, MRO, ARO, WRO) have TSH receptors but did not respond to TSH in regard to iodide uptake and thymidine incorporation. Both human thyroid carcinoma cells and FRTL-5 cells contain specific receptors for TNF. Scatchard analysis showed that the receptor numbers and dissociation constants in human thyroid carcinoma cells and FRTL-5 cells were, respectively; 2.4 x 10(4), 5.4 nM (WRO); 8 x 10(3), 3.4 nM (MRO); 4 x 10(3), 1 nM (ARO); 7 x 10(3), 1 nM (NPA); 3 x 10(3), 1 nM (FRTL-5), and 9 x 10(3), 1 nM (FRTL-5 cells treated with TSH). The results indicate that TNF affects thyroid cell function through binding to the TNF receptor and that the number of TNF receptors is regulated by TSH. PMID- 2551629 TI - Direct modulation by androgens of the response of human bone cells (SaOS-2) to human parathyroid hormone (PTH) and PTH-related protein. AB - We have reported previously that 17 beta-estradiol (E2) inhibits selectively the cAMP response to human (h) PTH and PTH-related protein (hPTHrP), but not to vasoactive intestinal peptide, in human osteoblast-like cells (SaOS-2). We have now extended these studies to investigate the actions of androgens on hPTH stimulated accumulation of cAMP, and on the roles of new protein synthesis and pertussis toxin (PTox) substrates in the actions of steroid hormones on SaOS-2 cells. Pretreatment with testosterone (T) or 5 alpha-dihydrotestosterone (5 alpha DHT) for 4-12 h at concentrations of 10(-12) to 10(-8) M inhibited significantly the cAMP response to hPTH by up to 50-70% of control. Like E2, the actions of T and 5 alpha-DHT were selective for hPTH or hPTHrP; there was no inhibition of the stimulatory action of vasoactive intestinal peptide. Two related steroids, 5 beta DHT and 17 alpha-epitestosterone, did not inhibit the action of hPTH. Pretreatment of cells with cycloheximide, under conditions which inhibited protein synthesis by greater than 90%, reduced the cAMP response to hPTH but did not block the further inhibitory actions of E2, T, or 5 alpha-DHT. Pretreamtent of cells with PTox (100 ng/ml) for 24 h, enhanced the accumulation of cAMP stimulated by hPTH consistent with an action of PTox on Gi; however, the inhibitory actions of E2, T, and 5 alpha-DHT on PTH-stimulated cAMP accumulation were not attenuated by PTox. We conclude that androgens, as well as estrogens, act directly on human bone cells to modulate selectively an early effect of hPTH. The inhibitory actions of these steroid hormones do not appear to depend on new protein synthesis and may not involve a functionally active PTox substrate, presumably Gi. PMID- 2551631 TI - Heterogeneity between brain and pituitary corticotropin-releasing factor receptors is due to differential glycosylation. AB - Chemical affinity cross-linking studies have identified brain and pituitary CRF receptors with similar pharmacological characteristics but different mol wts (anterior pituitary, 75,000; brain, 58,000). In order to determine whether the heterogeneous nature of CRF receptors was inherent in the protein, we examined the glycoprotein nature of both types of CRF receptors using lectin affinity chromatography and treatments with exo- and endoglycosidases. CRF receptors in both the cerebral cortex and anterior pituitary adsorbed to and specifically eluted from Concanavalin-A- and wheat germ agglutinin-immobilized lectin affinity columns, indicating that both forms of the receptor are glycoproteins containing complex and high-mannose carbohydrate moieties. Cerebral cortical CRF receptors were sensitive to both neuraminidase and alpha-mannosidase treatment while pituitary CRF receptors were only affected by neuraminidase treatment, suggesting that CRF receptors in brain and pituitary differed slightly in the nature of their glycosylation units. After treatment of cerebral cortical or anterior pituitary CRF receptors with the endoglycosidase, N-glycanase, the mol wts were markedly decreased; the mol wt of the anterior pituitary CRF receptor was decreased from 75,000 to approximately 40,000-45,000 while in a corresponding manner, the cortical receptor was decreased from 58,000 to approximately 40,000 45,000. Limited proteolysis after deglycosylation with N-glycanase using the proteinases Staphylococcus aureus V8 (S. aureus V8) or papain, generated virtually identical peptide fragments from anterior pituitary- or cerebral cortex labeled CRF receptor proteins. In summary, these data support the hypothesis that the ligand binding subunit of the CRF receptor in both brain and pituitary resides on a polypeptide of 40,000-45,000 and appears to be identical in both tissues. Differences observed in the mobility of the two proteins were found to be due to differences in the posttranslational modification of the proteins in the two tissues. PMID- 2551630 TI - Fentanyl stimulates prolactin release through mu-opiate receptors, but not the serotonergic system. AB - Both serotonin (5-HT) and opiates exert a stimulatory effect on PRL secretion. Some evidence suggests that the action of opiates may be elicited through serotonergic neurons. We tested this hypothesis in the present study by evaluating the effect of perturbation of the serotonergic system on PRL secretion induced by fentanyl, a potent morphine-like analgesic. Female Sprague-Dawley rats ovariectomized for 3 weeks and given polyestradiol phosphate (0.1 mg/rat) for 1 week were used in the study. Fentanyl, at a dose of 20 micrograms/rat, induced significant PRL secretion that peaked at 10 min and lasted for more than 30 min. Pretreatment with naloxone (0.5 mg/kg BW, ip) did not block the acute phase of PRL secretion, but significantly lowered the PRL level at 30 min. Fentanyl at a smaller dose (5 micrograms/rat, iv) still induced significant PRL release 10, but not 30, min after injection. This effect was significantly blocked by pretreatment with the same dose of naloxone. On the other hand, whereas animals pretreated with ketanserin or LY53857 (both at a dose of 5 mg/kg BW, ip), two specific 5-HT2 receptor antagonists, had no effect on fentanyl-induced PRL secretion, the same treatment significantly blocked 5-HT-induced PRL secretion. Likewise, pretreatment with p-chlorophenylalanine (250 mg/kg BW, ip), a 5-HT synthesis inhibitor, for 2 days had no effect on the action of fentanyl, while 5 HT-induced PRL secretion was significantly augmented. We conclude that fentanyl acts through mu-opiate receptors to stimulate PRL secretion in a process that does not involve the serotonergic system. PMID- 2551632 TI - Kinetic actions and interactions of arginine vasopressin, angiotensin-II, and oxytocin on adrenocorticotropin secretion by rat anterior pituitary cells in the microperifusion system. AB - We have examined the actions and interactions of arginine vasopressin (AVP), angiotensin-II (AII), and oxytocin (OT) on the ACTH secretory response of dispersed rat anterior pituitary cells in a microperifusion system. There was a dose-dependent ACTH secretory response to a 3-min perifusion of AII which reached its maximum 10 sec after the cells were exposed to AII and fell rapidly to baseline within 2 min, despite continued infusion of AII. This brief spike type of pattern is similar to that produced by AVP, but different from the sustained plateau response induced by CRF. The threshold stimulating concentration of AII was about 10(-9) M; the maximally stimulating concentration was not defined, but was 10(-6) M or more. The initial ACTH response to OT was similar, but fell to a plateau 2 min after the cells were exposed to OT and remained constant until perifusion with OT was stopped, after which it fell rapidly to baseline. The threshold stimulating concentration of OT was 10(-8) M; the maximally stimulating concentration was not defined, but was 10(-6) M or more. The ACTH secretory response to 10(-8) M AII was greatly diminished when cells were exposed to 10(-6) AVP or 10(-6) M OT before AII infusion. However, prior exposure to AII had no effect on the magnitude of the ACTH secretory response to either AVP or OT. The effects of simultaneous perifusion of AII and AVP and of AII and OT were additive. When AVP and OT were perifused sequentially, the ACTH secretory response to the peptide that was infused second was completely abolished. Furthermore, the combination of AVP and OT stimulated no greater response than either agent alone. When cells were perifused with the combination of 10(-7) M OT and 10(-7)- to 10(-5)-M concentrations of two potent AVP V1 receptor antagonists, [1-(beta-mercapto-beta,beta-cyclopentamethylenepropionic acid),2-(O methyl)tyrosine]-Arg8-vasopressin and [1-deaminopenicillamine-2-(O methyl)tyrosine]-Arg8-vasopressin, both phases of the response to OT were progressively and almost completely inhibited. The initial spike phase was inhibited at lower antagonist concentrations than the sustained plateau phase.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2551634 TI - T- and L-calcium channels in steroid-producing chicken granulosa cells in primary culture. AB - Steroidogenesis and other functions in granulosa cells are calcium dependent. Using the patch-clamp technique to record single ion channel activity, we have identified for the first time two kinds of calcium channels through which the divalent ion may enter chicken granulosa cells. The cells were maintained in primary culture whose basal and hormone-stimulated progesterone production was evaluated at different times in culture and at different temperatures. A small channel, with a conductance of 4-5 picosiemens (pS), had short openings and inactivated rapidly. A large channel had a conductance of 20-30 pS, a high activation threshold, and slow inactivation kinetics. The dihydropyridine compound Bay K-8644, a L-calcium channel agonist, significantly increased the activity of the large channel, and nifedipine, a dihydropyridine calcium channel blocker, inhibited it completely. Both types of channels were seen in functionally competent signal-responsive granulosa cells cultured for up to 48 h. Whether these channels are involved in steroidogenesis, protein production, and/or secretion remains to be established. PMID- 2551633 TI - The role of angiotensin-II in progressive diabetic glomerulopathy in the rat. AB - The potential pathogenic role of angiotensin-II (AII) in early progressive diabetic and renal ablation-induced glomerulosclerosis was explored and compared in the Sprague-Dawley (SD) rat and the mongrel dog. Male SD rats were divided into control and streptozotocin-treated (65 mg/kg, iv) groups. Unilateral surgical nephrectomy (Nx) was performed in half of each group. Enalapril (E; 50 mg/liter in the drinking water) was administered to half of each subgroup. Enalapril (high E; 250 mg/liter) was given to another 13 streptozotocin rats. All diabetic rats were treated with sc NPH insulin (4 U/day), and blood glucose was 520 +/- 124 mg/dl (mean +/- SD). Microalbuminuria was measured by RIA in 24-h urine collections; wet kidney weights were compared. [125I]AII binding assays were performed on isolated glomeruli. In control rats the high affinity binding dissociation constant (Kd) was 0.59 +/- 0.15 nM (n = 26; mean +/- SD) and receptor number (Ro) was 732 +/- 195 fmol/mg glomerular protein. At 3 weeks, the diabetic glomerular AII receptor Kd was 0.38 +/- 0.07 nM (n = 6; P less than 0.02 vs. control) and Ro was 784 +/- 97 fmol/mg protein (P = NS vs. control); diabetic high E Kd was 0.39 +/- 0.06 nM (n = 6; P less than 0.02 vs. control), and Ro was 873 +/- 105 fmol/mg protein (P = NS vs. diabetes without E). By 10 weeks, a Kd of 0.49 +/- 0.12 nM (n = 32; P less than 0.01 vs. control) and a Ro of 780 +/- 174 fmol/mg protein (P = NS vs. control) were observed when all of the diabetic group data were pooled. Neither Nx nor low or high dose E altered Ro. This is evidence that intraglomerular AII levels are normal or reduced after diabetes, Nx, or both. In the diabetic group, low dose E partially prevented, and high E abolished, Nx-enhanced microalbuminuria and renal hypertrophy. In nine pancreatectomized insulin-treated mongrel dogs over a 12- to 24-month period, despite moderately poor glucose control (300 +/- 75 mg/dl) and combined unilateral Nx in five dogs (12 months), elevated microalbuminuria was not observed. [125I]AII binding to isolated normal and diabetic dog glomeruli revealed the Kd to be of low affinity (1.86 +/- 0.28 to 13.80 +/- 1.88 nM), identifying the presence of type B receptors. Hence, the SD rat and mongrel dog differ in susceptibility to glomerular AII receptor type and progressive diabetic glomerulopathy.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2551635 TI - Pharmacological characterization of the voltage-dependent calcium channel of pancreatic B-cell. AB - Pharmacological characteristics of the voltage-dependent calcium channel (VDCC) of the pancreatic B-cell were studied using omega-conotoxin (omega CgTX) and dihydropyridine (DHP) calcium channel blockers. High glucose and potassium (K+) depolarization were employed as the stimulant of insulin release. omega CgTX (greater than 50 nM), a blocker of neural, but not muscular, Ca2+ channels, partially blocked (27%) the second, but not the first, phase of glucose-induced insulin release without a significant effect on K+ depolarization-induced insulin release. The DHP Ca2+ channel blocker nifedipine inhibited both phases of glucose induced insulin release (ED50 = 200 nM) and completely abolished both phases of response at 10 microM. In contrast, the DHP Ca2+ channel blocker only partially suppressed (75% at 10 microM) K+ depolarization-induced insulin release with an ED50 of 100 nM. We conclude that pancreatic B-cell possesses at least two classes of VDCCs; one is DHP sensitive, and the other DHP insensitive. Partial suppression of the second phase of glucose-induced insulin release by a high concentration of omega CgTX may be due to its toxic effect on the secretory machinery other than VDCC. PMID- 2551636 TI - Effects of dexamethasone and vitamin D3 on cartilage differentiation in a clonal chondrogenic cell population. AB - We have investigated the regulation of chondroblast/chondrocyte differentiation using a unique clonal cell population, designated RCJ 3.1C5 (C5), which differentiates into discrete three-dimensional cartilage nodules when grown in the presence of 15% fetal calf serum. Histologically, the nodules resembled hyaline cartilage; they contained large rounded chondrocytes surrounded by a refractile matrix which stained intensely with Alcian blue, exhibited metachromasia after Toluidine blue staining, and stained with an antibody against type II collagen. The cartilage nodules that formed did not mineralize, despite the presence of organic phosphate in the culture medium. The synthetic glucocorticoid dexamethasone (DEX) increased the number of cartilage nodules formed in a dose-dependent manner (ED50, approximately 10(-9) M), with a maximal stimulatory dose of 10(-8) M. DEX had no effect on the population doubling time and saturation density. The effects of DEX on the number of cartilage nodules were similar whether it was added from the beginning of the culture period (starting during exponential growth) or at confluence. In contrast, 1,25 dihydroxyvitamin D3 [1,25-(OH)2D3] inhibited cartilage nodule formation in a dose dependent manner (IC50, approximately 5 x 10(-10) M), with maximum inhibition at 10(-7) M. In addition, 1,25-(OH)2D3 decreased cell proliferation and saturation density. Equimolar doses of the vitamin D3 metabolites 24,25-dihydroxyvitamin D3 and 25-hydroxyvitamin D3 had no effect. C5 cells treated with 1,25-(OH)2D3 in the absence of DEX during the exponential growth phase exhibited a reduced capacity to form cartilage nodules upon subsequent exposure to DEX. At confluence, before cartilage nodules had formed, C5 cells responded to PTH and prostaglandin-E2 with increases in intracellular cAMP of about 10- and 95-fold respectively. After cartilage nodules were present, prostaglandin-E2 responsiveness decreased to about 25-fold, whereas there was no significant change in PTH responsiveness. DEX decreased the population alkaline phosphatase levels at all times measured, whereas 1,25-(OH)2D3 had a biphasic effect: an increase at 5 days in culture, followed by a decrease at later times in culture. These data indicate that the clonal cell line RCJ 3.1C5 is a useful model system in which to investigate cartilage differentiation. PMID- 2551637 TI - Insulin and insulin-like growth factor-I stimulate the 3 beta-hydroxysteroid dehydrogenase activity of human placental cytotrophoblasts. AB - The placenta is the primary source of progesterone during pregnancy. Because pregnant diabetic women are reported to have higher serum progesterone levels than nondiabetic pregnant women, we studied the roles of insulin and insulin-like growth factor-I (IGF-I) in the regulation of human cytotrophoblastic 3 beta hydroxysteroid dehydrogenase (3 beta HSD) activity. Incubation of cytotrophoblasts with insulin or IGF-I for 24 h significantly increased the ability of these cells to convert pregnenolone to progesterone by 75.8 +/- 16.5% (+/- SE) and 65.4 +/- 12.7%, respectively. Treatment with either insulin or IGF-I did not alter cytotrophoblastic production of 20 alpha-hydroxypregn-4-en-3-one (the primary metabolite of progesterone), thus demonstrating that these peptides increased progesterone synthesis (i.e. 3 beta HSD activity) rather than decreased progesterone catabolism. Insulin and IGF-I stimulated 3 beta HSD activity at concentrations as low as 50 and 10 ng/ml, respectively. Insulin- and IGF-I stimulated 3 beta HSD activities were completely inhibited by concurrent treatment with either actinomycin-D or cycloheximide, suggesting that new mRNA and protein synthesis are required for these peptides to exert their effects. Blockade of the IGF-I receptor of cytotrophoblasts with alpha IR-3, a monoclonal anti-IGF-I receptor antibody, prevented the stimulation of 3 beta HSD activity by IGF-I, but did not alter insulin's stimulatory effect. Thus, the two hormones can each stimulate 3 beta HSD activity via activation of their respective receptors. These studies indicate that insulin and IGF-I can regulate human cytotrophoblastic 3 beta HSD activity in vitro. Since pregnant diabetic women manifest peripheral hyperinsulinemia, and IGF-I levels in fetal cord sera from diabetic pregnancies may be elevated, these observations may help explain the elevated serum progesterone levels associated with pregnancy in the diabetic patient. PMID- 2551638 TI - Regulation of Leydig cell function in primary culture by inhibin and activin. AB - Inhibin and activin are gonadal glycoproteins that selectively inhibit and stimulate FSH release, respectively. Previously we have reported that transforming growth factor-beta inhibited hCG-stimulated testosterone formation in mature Leydig cells. In the present study we evaluated the effects of other members of the transforming growth factor-beta family, inhibin and activin, on Leydig cell function. We found that activin (0.1-10 ng/ml) had no effect on basal testosterone formation, but inhibited hCG-stimulated testosterone formation in a dose-dependent manner. Activin (10 ng/ml) inhibited hCG-stimulated testosterone formation by 42%. Activin also inhibited hCG-stimulated cAMP formation. In the presence of activin (5 ng/ml), forskolin (10 microM)- and 8-bromo-cAMP (0.1 mM) induced testosterone formation were reduced about one third. Conversions of pregnenolone and progesterone to testosterone were also blocked by activin. Interestingly, [125I]hCG binding to Leydig cells and forskolin-induced cAMP formation were not affected by the addition of activin. In contrast to activin, inhibin (0.1-10 ng/ml) had no effect on hCG-induced testosterone formation at any concentration used. However, the inhibitory effects of activin on Leydig cell function were reversed by the concomitant addition of inhibin. Our results suggest that activin inhibits testosterone formation by the Leydig cells derived from normal mature rats. Multiple steps of the steroidogenic pathway are affected by testosterone. Inhibin alone has no effect, but reverses the inhibitory action of activin. PMID- 2551639 TI - Adenosine stimulates adenosine 3',5'-monophosphate and guanosine 3',5' monophosphate accumulation in rat pinealocytes: evidence for a role for adenosine in pineal neurotransmission. AB - Adenosine produces a concentration-dependent increase in pinealocyte cAMP (EC50, approximately 0.3 nM) and cGMP accumulation (EC50, approximately 0.7 nM). Maximal increases in both nucleotides are evident 10 min after treatment; 1 h later values return to pretreatment levels. Concentration-dependent effects on cAMP are also observed with N6-(L-2-phenylisopropyl)adenosine (EC50, approximately 0.75 nM), 5'-N-ethylcarboxy aminoadenosine (EC50, approximately 0.75 nM), and 2 chloroadenosine (EC50, approximately 2.0 nM); the EC50 values for stimulation of cGMP with these agents are higher by a factor of 2-10. In the case of 5'-N ethylcarboxy amidoadenosine, the concentration-response curve is biphasic, with a significant effect evident within the range of 1-100 pM. The stimulatory nature of this response and the relative potency of the agonists tested are consistent with the involvement of an A2-like adenosine receptor. Comparison of adenosine and the selective beta-adrenergic agonist isoproterenol indicated that their maximal EC50 values were generally similar. Studies with antagonists revealed that both 8-(p-sulfophenyl)theophylline (1 microM) and the xanthine amine congener (8-[4-[[[(2-aminoethyl)carbonyl]methyl]oxy]phenyl]1,3- dipropylxanthine (1 microM) inhibited the effects of adenosine (1 nM to 1 microM), but xanthine amine congener was more potent; the latter was markedly effective at 0.1 nM, whereas 8-(p-sulfophenyl)theophylline was nearly ineffective at this concentration. It was also determined that pineal cells generate extracellular adenosine from extracellular ATP. ATP is thought to be released along with catecholamines during neurotransmission. Hence, these studies support the view that adenosine could participate in the transsynaptic regulation of pineal function. PMID- 2551640 TI - The human liver growth hormone receptor. AB - Human livers, obtained from donors at the time of transplant, were homogenized in 0.25 M sucrose and fractionated by differential centrifugation. The specific binding of [125I] human (h) GH to total particulate fractions from 18 livers varied from 0.4-5.1% of the total radioactivity/100 micrograms protein. Binding affinity was 2.0 +/- 0.3 X 10(9) M-1, and binding capacity ranged from 14-53 fmol/mg protein. A different proportion of receptors occupied by endogenous hGH did not explain the large variation in binding. Binding sites were specific for hGH. Dissociation of the hormone-receptor complex was extremely slow. No specific binding of [125I]hPRL was observed. Specific binding of insulin was found in fractions from all livers and varied less than hGH binding. Cross-linking of [125I]hGH to plasma membrane and microsome receptors yielded two major autoradiographic bands corresponding to an estimated mol wt of 103,000 for the receptor, with a possible subunit of 54,000. Human liver primary fractions were characterized. The binding of hGH and insulin displayed a nucleo-microsomal distribution pattern in the primary fractions; 54.2% and 27.9% of the hGH-binding activity were found in the microsomes and the nuclear fraction, respectively, whereas insulin binds equally to nuclear and microsomal elements. Our findings suggest that hGH-binding sites are present in the plasma membrane and also in one or more intracellular compartments, whereas a high proportion of insulin receptors is associated with the plasma membrane. PMID- 2551641 TI - The posterior pituitary regulates prolactin, but not adrenocorticotropin or gonadotropin, secretion in the sheep. AB - The aim of this study was to determine the role of the posterior pituitary gland in the control of PRL, LH, FSH, and ACTH secretion in sheep. Posterior pituitary function was removed in ovariectomized ewes by electrical lesioning of the hypothalamo-neurohypophysial tract immediately posterior to the stalk-median eminence (LESION); controls were subjected to sham surgery (SHAM). LESION caused a 2-fold increase in plasma PRL concentrations on days 1-3 after surgery. Thereafter, concentrations gradually declined until they were similar to those in SHAM ewes. There was no change in plasma concentrations of LH, FSH, or ACTH after LESION. Plasma PRL responses to insulin in SHAM ewes were completely abolished, and the plasma PRL response to chlorpromazine was reduced to almost half by LESION. In contrast, audiovisual stress (barking dog) and serotonin challenge caused an immediate release of PRL in both LESION and SHAM ewes, with the amplitude of the responses indistinguishable between groups. LESION had no effect on the plasma ACTH responses to audiovisual stress, insulin, or serotonin. We conclude that the posterior pituitary gland is involved in the regulation of PRL under some circumstances, but not of LH, FSH, or ACTH secretion in the sheep. Accordingly, changes in PRL release after hypothalamopituitary disconnection in this species may reflect a loss of posterior lobe function rather than the removal of hypothalamic inputs. In addition, the PRL response to insulin is dependent on a functional posterior pituitary gland, whereas responses to audiovisual stress and serotonin appear to rely on inputs to the pituitary gland via the median eminence and the long hypothalamo-hypophysial portal blood vessels. PMID- 2551642 TI - Non-homologous sequences of parathyroid hormone and the parathyroid hormone related peptide bind to a common receptor on ROS 17/2.8 cells. AB - We and others have recently shown that amino terminal sequences of parathyroid hormone (PTH) and parathyroid hormone related peptide (PTHrP), which share a 62% homology within the first 13 residues, bind to the same receptor on ROS 17/2.8 cells. The remaining PTHrP sequence is markedly different from PTH, suggesting that receptor binding may be dependent on the first 13 amino acids of either peptide. However, since the amino acid residues 14-34 have previously been recognized as an important binding domain for PTH, conformational similarity within this portion's secondary structure of both peptides could contribute to their capacity to bind to the same receptor. To test this hypothesis, we synthesized [Tyr36,Cys38]PTHrP-(14-38) and [Tyr34]bPTH(14-34)NH2, and studied binding of both peptides to the common PTH/PTHrP receptor on ROS 17/2.8 cells. Radioiodinated, HPLC-purified [Nle8,18, Tyr34]bPTH(1-34)NH2 (NlePTH) and [Tyr36]PTHrP-(1-36)NH2 were used to functionally define receptor binding requirements. [Tyr36,Cys38]PTHrP(14-38) and [Tyr34]bPTH(14-34)NH2 competed with 125I-NlePTH for binding sites on ROS 17/2.8 cells with apparent Kds of 10 microM and 50 microM respectively. Both peptides also competed with 125I-[Tyr36]PTHrP(1 36)NH2 with apparent Kds of 30 microM and 10 microM respectively. In the same assay system, NlePTH and [Tyr36,Cys38]PTHrP(1-38)inhibited binding of either radioiodinated ligand with apparent Kds of 0.3 and 1.0 nM. These studies indicate that although [Tyr34]bPTH(14-34)NH2 and [Tyr36,Cys38]PTHrP(14-38) share virtually no sequence homology, their secondary structures must be sufficiently similar to permit binding to a common PTH/PTHrP receptor. PMID- 2551643 TI - Vascular type I aldosterone binding sites are physiological mineralocorticoid receptors. AB - In vitro, Type I receptors have high and equivalent affinity for aldosterone, corticosterone and cortisol: in vivo, physiological mineralocorticoid target tissues (kidney, colon, parotid) are highly aldosterone-selective, in contrast with hippocampus and heart. In the present study we show that the mesenteric vascular arcade is similarly highly aldosterone-selective in vivo, and in vitro shows considerable levels of 11 beta OH steroid dehydrogenase activity, previously postulated as the mechanism whereby glucocorticoids are excluded from physiological mineralocorticoid receptors. PMID- 2551644 TI - Y-1 adrenocortical tumor cells contain atrial natriuretic peptide receptors which regulate cyclic nucleotide metabolism and steroidogenesis. AB - The effects of atrial natriuretic peptide (ANP) on adrenocortical fasciculata cells were examined in the ACTH-responsive Y-1 mouse adrenocortical tumor cell line. Y-1 cell membranes rapidly bound [125I]ANP, with equilibrium binding (22 C) reached within 45 min. Binding of [125I]ANP was inhibited in a concentration dependent manner by unlabeled ANP and atriopeptin-I (IC50, approximately 1.2 X 10(-9) and 1.6 X 10(-8) M, respectively), but not by C- or N-terminal-deleted ANP fragments, ACTH, or arginine vasopressin (up to 10(-6) M). Scatchard analysis revealed a single class of high affinity binding sites with a Kd of 1.6 X 10(-10) M and a binding capacity of 560 fmol/mg protein. Photo-affinity labeling demonstrated the specific binding of ANP to two protein entities of 130 and 63 kDa. ANP stimulated both cGMP synthesis and secretion from Y-1 cells (EC50, approximately 3.5 X 10(-9) M). Release of the nucleotide was inhibited by probenecid (IC50, approximately 5 X 10(-5) M). The atrial peptide partially inhibited ACTH-stimulated cAMP formation (IC50, approximately 10(-8) M) and partially antagonized basal and ACTH-stimulated steroidogenesis. The data demonstrate the presence in Y-1 cells of specific and saturable ANP receptors, activation of which leads to changes in cyclic nucleotide metabolism and inhibition of steroidogenesis. PMID- 2551645 TI - Recombinant mouse placental lactogen-I binds to lactogen receptors in mouse liver and ovary: partial characterization of the ovarian receptor. AB - The binding of recombinant mouse placental lactogen-I (mPL-Ir) to liver and ovarian membranes was investigated in virgin and pregnant mice. Competitive binding assays demonstrated that mPL-Ir, mouse placental lactogen-II (mPL-II), and mouse PRL (mPRL) bind to the same receptors in ovarian membranes. The relative abilities of the three hormones to displace [125I]iodo-mPL-Ir from the ovarian lactogen receptors was mPL-II greater than mPL-Ir much greater than mPRL. Scatchard analysis of mPL-Ir binding to ovarian membranes from day 10 pregnant mice showed a Ka of 2.0 x 10(9) M-1 and a binding capacity of 3.2 x 10(-14) mol/mg membrane protein. The specific binding of [125I]iodo-mPL-Ir to ovarian membrane preparations was significantly higher on day 17 than on day 10 of gestation. Dissociation of endogenous hormones with 4 M MgCl2 increased the binding of [125I]iodo-mPL-Ir to ovarian membranes but not to liver membranes. Affinity cross-linking of [125I]iodo-mPL-Ir to liver and ovarian membranes resulted in the specific labeling of proteins with receptor mol wt (Mr) of 44K and 40K (nonreduced) and 50K and 42K (reduced), as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The lactogen receptors from liver and ovary appeared structurally homologous, producing fragments with similar Mr when treated with proteolytic enzymes and undergoing similar reductions in Mr when treated with glycolytic enzymes. The ability of mPLs to bind specifically and with high affinity to receptors in mouse ovarian membranes suggests that these hormones may regulate ovarian function during gestation. PMID- 2551646 TI - Development of receptors for insulin and insulin-like growth factor-I in head and brain of chick embryos: autoradiographic localization. AB - In whole brain of chick embryos insulin receptors are highest at the end of embryonic development, while insulin-like growth factor-I (IGF-I) receptors dominate in the early stages. These studies provided evidence for developmental regulation of both types of receptors, but they did not provide information on possible differences between brain regions at each developmental stage or within one region at different embryonic ages. We have now localized the specific binding of [125I]insulin and [125I]IGF-I in sections of head and brain using autoradiography and computer-assisted densitometric analysis. Embryos have been studied from the latter part of organogenesis (days 6 and 12) through late development (day 18, i.e. 3 days before hatching), and the binding patterns have been compared with those in the adult brain. At all ages the binding of both ligands was to discrete anatomical regions. Interestingly, while in late embryos and adult brain the patterns of [125I]insulin and [125I] IGF-I binding were quite distinct, in young embryos both ligands showed very similar localization of binding. In young embryos the retina and lateral wall of the growing encephalic vesicles had the highest binding of both [125I]insulin and [125I]IGF-I. In older embryos, as in the adult brain, insulin binding was high in the paleostriatum augmentatum and molecular layer of the cerebellum, while IGF-I binding was prominent in the hippocampus and neostriatum. The mapping of receptors in a vertebrate embryo model from early prenatal development until adulthood predicts great overlap in any possible function of insulin and IGF-I in brain development, while it anticipates differential localized actions of the peptides in the mature brain. PMID- 2551647 TI - Mechanism of insulin resistance induced by sustained levels of cytosolic free calcium in rat adipocytes. AB - We have recently provided evidence that elevated levels of cytosolic free Ca2+ ([Ca2+]i) decreased insulin-stimulated glucose uptake in isolated rat adipocytes. To investigate the mechanism of Ca2+ action, we examined the effects of elevated levels of [Ca2+]i on insulin binding, autophosphorylation, and tyrosine kinase activity (TKA) of insulin receptors as well as basal and insulin-stimulated cellular distribution of glucose transporters. The latter was assessed by cytochalasin-B binding to plasma membrane and cytosolic fractions. Elevated concentrations of [Ca2+]i were maintained by incubating adipocytes with a depolarizing concentration of K+ (40 mM). Basal nonstimulated glucose uptake was not altered by increased levels of [Ca2+]i. Adipocytes with higher [Ca2+]i (220 +/- 15 nM) showed 30% reduction in insulin-stimulated 2-deoxyglucose uptake compared with control cells ([Ca2+]i, 140 +/- 18 nM). Moreover, adipocytes with higher levels of [Ca2+]i demonstrated an approximately 10% reduction in autophosphorylation and TKA of insulin receptors without a change in insulin binding. Both basal and insulin-stimulated distributions of glucose transporters were unaffected by sustained levels of [Ca2+]i. The effects of elevated [Ca2+]i were not mimicked by protein kinase-C activation. These observations suggest that 1) elevated or sustained levels of [Ca2+]i impair insulin-stimulated glucose uptake; and 2) Ca2+-induced impairment appears to reside at the postbinding steps of insulin action and probably interferes with the TKA of insulin receptors and the intrinsic activity of glucose transporters. PMID- 2551649 TI - Facilitative actions of the protein kinase-C effector system on hormonally stimulated adenosine 3',5'-monophosphate production by swine luteal cells. AB - The exact nature of the interaction(s) between cAMP and calcium-sensitive phospholipid-dependent protein kinase-C effector pathways is not well understood in many tissues, including the ovary. In the present work we have evaluated the ability of protein kinase-C to modulate receptor-and nonreceptor-mediated cAMP generation in acute suspension cultures of swine luteal cells. Cells were exposed to LH (1 micrograms/ml), forskolin (100 microM), cholera toxin (1 microgram/ml), pertussis toxin (100 ng/ml), and/or phorbol ester [12-O-tetradecanoylphorbol-13 acetate (TPA)] for 0-90 min. TPA had no effect on basal cAMP accumulation, but increased (P less than 0.05) LH-, forskolin-, and cholera toxin-activated cAMP formation, with maximal facilitation at 30, 45, and 60 min, respectively. This facilitative effect was robust, as it could be demonstrated in both the presence and absence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (0.5 mM). TPA increased dose-dependent LH (0.1-1 microgram/ml)-, forskolin (3-300 microM-, and cholera toxin (0.3-10 microgram/ml)-stimulated cAMP accumulation. TPA induced a dose-dependent (0.3-30 ng/ml) increase in cAMP accumulation when incubated with the half-maximally effective (ED50) and maximally effective doses of LH (0.8 and 1 microgram/ml, respectively), forskolin (10 and 300 microM), and cholera toxin (0.2 and 3 micrograms/ml). TPA had an ED50 for this functional activation of 6.1 (67% confidence interval, 4.4-9.7) nM. The stimulatory effect of TPA could be mimicked by two synthetic diacylglycerols, 1,2-Dioctanoylglycerol and 1-oleoyl-2-acetylglycerol, but not by inactive phorbol esters. In addition, TPA augmented the stimulatory effect of pertussis toxin when combined with maximally effective doses of LH, forskolin, and cholera toxin. The stimulatory action of TPA on cAMP production was limited to endogenous cellular adenylyl cyclase. Bacterially derived adenylyl cyclase toxin isolated from Bordetella pertussis resulted in a dose-dependent increase in cAMP formation over 60 min, which was not facilitated by phorbol ester. We conclude that stimulatory coupling exists between the calcium-dependent protein kinase-C and cAMP-generating systems in swine luteal cells. This stimulatory coupling is enacted in part at the levels of both the guanine binding and the catalytic subunits of adenylyl cyclase. PMID- 2551648 TI - Localization and structural characterization of insulin-like growth factor receptors in mammalian retina. AB - Insulin-like growth factors (IGFs) are peptide mitogens, structurally related to insulin, whose biological actions in the CNS are incompletely known. The retina is largely uncharacterized with respect to IGF receptors. We, therefore, studied IGF receptors in bovine and murine retinal tissues by immunohistochemistry, autoradiographic localization, and affinity labeling. Notable IGF-II receptor immunoreactivity was found in retinal pigment epithelium (RPE), with intermediate levels in choroid, low levels in the inner and outer plexiform layers and outer nuclear layer, and very low levels in other regions. Autoradiographic localization using [125I]IGF-II confirmed the IGF-II receptor immunohistochemistry. Autoradiographic localization using [125I]IGF-I labeled the nuclear layers and the photoreceptor region. Affinity labeling disclosed differences in the apparent mol wt of IGF-I and IGF-II receptors from bovine eye tissues and those from liver and brain. IGF-I receptor alpha-subunits (the IGF binding subunit) migrated at: liver, 139,000; brain, 125,000; RPE, 125,000 and 135,000 (two sizes); and retina, 125,000 and 135,000. IGF-II receptors migrated at: liver, 245,000; brain, 235,000; RPE, 240,000; and retina, 230,000. We conclude that mammalian retina contains both IGF-I and -II receptors, which differ from those found in other tissues and have a characteristic spatial distribution within the retina. PMID- 2551650 TI - Effect of diabetes on hormone-stimulated and basal hepatocyte calcium metabolism. AB - The effects of diabetes on basal calcium metabolism and the response to endocrine stimulation were studied in hepatocytes from acute and long term diabetic rats. Hepatocyte calcium sequestration and turnover were increased in both acute and chronic diabetes. Cytosolic free calcium (Cai2+) was significantly increased in the chronic diabetics, but the rise in Cai2+ evoked by epinephrine, angiotensin, vasopressin, and glucagon was depressed. The blunted stimulation of phosphorylase alpha activity in the diabetics was influenced by a 50-60% decrease in total cell activity of glycogen phosphorylase and the decreased rise in cytosolic free calcium. Insulin replacement corrected both basal and stimulated changes in the acute diabetes model. Depressed [3H]inositol trisphosphate formation in response to epinephrine or vasopressin and increased intracellular organelle calcium buffering were observed in hepatocytes from diabetic animals; both may effect the diminished rise in Cai2+. Several possible causes for the depressed rise in Cai2+ after stimulation in chronic diabetic animals were eliminated: 1) the number and affinity of alpha 1-adrenergic receptors for epinephrine were normal; 2) the initial rise in calcium influx evoked by epinephrine or vasopressin was not depressed; and 3) the ability of inositol trisphosphate to release calcium from intracellular organelles was not changed. The results suggest that the diabetic changes in calcium-mediated endocrine regulation of hepatic carbohydrate metabolism contribute to the general pathology of the disease. PMID- 2551651 TI - The effect of a 5 alpha-reductase inhibitor on androgen physiology in the immature male rat. AB - To provide insight into the role of 5 alpha-dihydrotestosterone (DHT) in postnatal androgen physiology, we administered the 5 alpha-reductase inhibitor finasteride to male rats from birth through the onset of puberty. In 4-week-old control rats serum testosterone levels averaged 0.21 ng/ml, and DHT levels averaged 0.64 ng/ml. By 7 weeks of age, testosterone levels increased more than 7 fold to 1.57 ng/ml, while the circulating DHT level declined to 0.26 ng/ml. In both the 4- and 7-week-old inhibitor-treated animals, circulating DHT levels were 25-50% of control values, and circulating testosterone levels were higher than control values. In 7-week-old inhibitor-treated rats, the weights of prostate, penis, seminal vesicles, and epididymal tissues were only 30-50% those of the controls. However, DHT formation is apparently not critical for postnatal development of the preputial glands or the androgen-dependent perineal muscles, since the weights of these tissues were not affected by treatment with inhibitor. Treatment with the 5 alpha-reductase inhibitor had no apparent effect on testicular histology or daily sperm production despite the fact that testicular DHT content was lower (70%) and testosterone content was higher (250%) than those in controls. We conclude that DHT formation is important for the normal postnatal growth of the prostate, seminal vesicles, epididymis, and penis and may be important for normal feedback control of testosterone production in rats, but that its formation is not critical for the onset of spermatogenesis or the development of the preputial glands or the androgen-dependent perineal muscles. PMID- 2551652 TI - The muscarinic cholinergic agonist arecoline stimulates the rat hypothalamic pituitary-adrenal axis through a centrally-mediated corticotropin-releasing hormone-dependent mechanism. AB - Several lines of experimental evidence suggest that acetylcholine and other cholinergic agonists are excitatory to the hypothalamic-pituitary-adrenal (HPA) axis. To examine the site on the HPA axis that is stimulated by cholinergic agents, we evaluated the in vivo and in vitro effects of the muscarinic cholinergic agonist arecoline in intact and pituitary stalk-transected rats as well as on isolated rat hypothalami, dispersed anterior pituicytes, and adrenocortical cells in culture. Arecoline, injected iv to catheterized, freely moving male Sprague-Dawley rats, stimulated plasma ACTH and corticosterone release in a dose-dependent fashion. The muscarinic cholinergic antagonist atropine significantly blunted the ACTH response to arecoline. Pituitary stalk transection led to diminished plasma ACTH and corticosterone responses to arecoline. Similarly, previous administration of anti-CRH serum significantly blunted these responses. These findings suggest that arecoline stimulates the HPA axis centrally, mainly via secretion of CRH. This hypothesis was confirmed by the dose-dependent ability of arecoline to cause hypothalamic CRH secretion in vitro, an effect antagonized by atropine, and its failure to elicit ACTH and corticosterone secretion by dispersed anterior pituicytes and adrenocortical cells in culture, respectively. These data suggest that the muscarinic cholinergic agonist arecoline stimulates the HPA axis in the rat and that this effect is mediated mainly by the release of endogenous CRH. Arecoline, therefore, appears to be a compound suitable to selectively evaluate the responsiveness of the central component of the HPA axis. PMID- 2551653 TI - Alterations in the hepatic insulin receptor kinase in genetic and acquired obesity in rats. AB - Obesity is associated with insulin resistance and type II diabetes mellitus. In the present study, we have characterized hepatic insulin receptor function in two animal models of obesity: the Zucker fatty rat (ZFR), a model of genetic obesity with severe hyperinsulinemia, and the Sprague-Dawley rat with dietary obesity, a model of acquired obesity. Zucker fatty rats were also treated with streptozotocin (STZ) in an effort to examine the effects of relative insulin deficiency and hyperglycemia in the setting of obesity. Using wheat germ agglutinin-purified insulin receptor extracted from liver, no significant difference in insulin binding was identified in either model of obesity. beta Subunit autophosphorylation was significantly decreased in both obese models relative to that in controls (72% in the obese ZFR and 49% in the overfed Sprague Dawley model). Kinase activity, as measured by phosphorylation of the 1142-1153 synthetic peptide, was also decreased in both models of obesity by 22% and 64%, respectively. In the Zucker rat, STZ treatment led to an 80% increase in receptor concentration and a further 70% increase in beta-subunit autophosphorylation per receptor, whereas tyrosine kinase activity toward substrate was not altered. Since kinase activity is closely linked to autophosphorylation, we determined the fraction of autophosphorylated (activated) receptors vs. non-phosphorylated (inactive) receptors by using antiphosphotyrosine antibody to precipitate receptors bound with [125I]insulin. There was no significant difference in the percentage of activated insulin receptors in the dietary obese, ZFR, or STZ treated Zucker rat vs. that in the controls. In all models, the percentage of activated receptors ranged from 32-46% of the total receptor pool. These data suggest that in genetic and acquired obesity, autophosphorylation of the beta subunit is reduced and is a limiting factor in insulin receptor activation. A similar fraction of all receptors appears to undergo some level of autophosphorylation; however, full autophosphorylation and, thus, activation of the receptor do not occur, and this results in a decrease in kinase activity. This block in autophosphorylation may account for significant reductions in insulin receptor kinase function in obesity. PMID- 2551654 TI - Localization, characterization, and quantification of insulin-like growth factor I-binding sites in the ewe ovary. AB - To assess a potential role of insulin-like growth factor-I (IGF-I) in the ewe ovary, the presence of IGF-I receptors and IGF-I-binding proteins was studied by binding assays performed on granulosa cell suspensions, in follicular fluid, and on ovarian sections. On the ovarian sections, labeling was quantified after autoradiography by microphotometry. Competition studies with IGF-I and insulin allowed us to estimate the relative proportions of binding proteins and type I receptors in the different compartments of the ewe ovary. Our results clearly show that saturable, specific, and high affinity IGF-I receptors are present on the ovine granulosa cells. At equilibrium for both granulosa cell suspensions and frozen sections, the Kd value was close to 2 nM. IGF-I binding proteins were also present in follicular fluid and stroma, thecal, and granulosa cells. At equilibrium for follicular fluid, the Kd value was 0.91 +/- 0.27 nM (mean +/- SE). Moreover, on frozen sections, it was shown that atresia of small follicles (less than 2 mm) was accompanied by a decrease in the number of IGF-I receptors and an increase in the number of IGF-I-binding proteins on granulosa cells. By contrast, this phenomenon was not observed in large follicles. These data indicate that granulosa cells of ewe ovary possess type I receptors, and IGF-I binding proteins may modulate IGF-I action in the process of follicular growth and atresia. PMID- 2551655 TI - Effects of atrial natriuretic factor on cyclic nucleotides, bone resorption, collagen and deoxyribonucleic acid synthesis, and prostaglandin E2 production in fetal rat bone cultures. AB - We examined the effects of synthetic human atrial natriuretic factor (human ANF 99-126) on adenylate cyclase activity, cAMP and cyclic GMP (cGMP) levels, bone resorption, collagen and DNA synthesis, and prostaglandin E2 (PGE2) production in fetal rat bone organ cultures. ANF (100 nM) inhibited PTH- and PGE2-stimulated cAMP production but had no effect on basal cAMP production in 21-day fetal rat calvaria. ANF increased cGMP levels, and this was not affected by PTH. ANF (10 nM) partially inhibited bone resorption stimulated by PGE2 but had no effect on control or PTH-stimulated resorption in 19-day fetal rat long bones. ANF had no effect on collagen and DNA synthesis or PGE2 production and did not alter responses to PTH or PGE2 in the fetal rat calvaria. Thus, ANF has no major direct effect on bone resorption or formation, but it is possible that ANF modulates the local regulatory function of PGE2 in bone. PMID- 2551656 TI - Growth and secretory responses of enriched populations of corticotropes. AB - The purpose of this study was to learn whether enriched populations of corticotropes could be grown without the other pituitary cell types. Corticotropes populations were enriched to 80-90% by counterflow centrifugation in an elutriator with the Sanderson chamber. After initial separation into small, medium, and large fractions, the cells were stimulated for 3 h with 0.5 nM corticotropin-releasing hormone (CRH) and then re-eluted to remove the enlarged corticotropes. More ACTH (6- to 10-fold) was released in media with 10% fetal bovine serum (FBS) than was released in media with no serum. The effects of FBS could not be mimicked by 0.3% BSA. Corticotropes grew in serum-containing media as long as they were plated at a density of at least 2500 cells per well. The corticotropes expanded in size and assumed two major morphological subtypes. Both stored ACTH and beta-endorphin. One subtype was flattened and pleomorphic. The other subtype was stellate with multiple processes. Cell counts showed a 2.5- to 3.8-fold increase in the number of labeled corticotropes during the first 21 days of culture. Then the numbers of cells declined rapidly. Basal secretion of ACTH rose 1.6-fold during the first week, plateaued after 14 days and then declined to less than 30% of first week levels. CRH stimulation produced dose-dependent increases in media ACTH. In 7 day cultures, both basal and stimulated levels of ACTH were similar to those in 7 day cultures of mixed pituitary cells (containing equivalent numbers of corticotropes). Stimulatory effects of CRH were evident for up to 42 days of culture. Arginine vasopressin enhanced CRH-mediated secretion in most cultures in the first week. Pretreatment with glucocorticoids (100 nM corticosterone) for 15 h blocked CRH-mediated secretion in all cultures. The studies showed that corticotropes do not need the other pituitary cell types for basic plating and basal and CRH-mediated secretory responses. Further tests of specific growth factors are needed to learn whether they will maintain function for longer periods. PMID- 2551657 TI - Thyrotropin-releasing factor-induced adrenocorticotropin secretion is mediated by corticotropin-releasing factor. AB - TSH-releasing factor (TRF), administered into the lateral cerebroventricle of adult male rats, elevated plasma concentrations of ACTH, epinephrine, and norepinephrine. TRF given iv was devoid of these activities. The CRF receptor antagonist, alpha-helical CRF9-41 (CRF9-41) given iv suppressed the TRF-induced increase in ACTH, but did not alter TRF-induced changes in plasma catecholamines. Intravenous administration of CRF antiserum totally blocked TRF-induced elevation of plasma ACTH concentrations. CRF receptor antagonists administered icv attenuated CRF-induced, but not TRF-induced elevation of plasma concentrations of ACTH, epinephrine, and norepinephrine. It is concluded from these results that TRF acts within the central nervous system to stimulate ACTH release through a CRF-dependent mechanism. PMID- 2551658 TI - Calcium currents in rat myoballs and their inhibition by insulin. AB - Two Ca2+ currents were found in myoballs prepared from primary culture of hindlimb muscles from rat embryos. One whole cell current, not described previously, was early, fast, and transient. It depended on the presence of Na+ in the bathing solution and was blocked by tetrodotoxin. Despite this behavior suggesting that it might be via a Na+ channel, its reversal potential exceeded 80 mV compared to 46 mV for the Na+ equilibrium potential. It was increased by increased Ca2+ concentration in the bathing solution and eliminated by Co2+ and by diltiazem. The other Ca2+ current resembled the slow inward Ca2+ current ICa(si), described in other cell types. Insulin decreased both Ca2+ currents; even at 64 pM insulin reduced ICa(si). When outward K+ currents were prevented, the myoball action potential was altered greatly, owing to the unopposed effect of ICa(si). Its duration was on the order of seconds. Insulin, in a concentration dependent manner, beginning at 60 pM, reduced the duration of this action potential more effectively than nimodipine. This is the most sensitive response to insulin observed in skeletal muscle. PMID- 2551659 TI - Oxytocin production and oxytocin messenger ribonucleic acid levels in bovine granulosa cells are regulated by insulin and insulin-like growth factor-I: dependence on developmental status of the ovarian follicle. AB - Oxytocin is a major peptide product of the ruminant corpus luteum, and the release of oxytocin from serum-free cultures of bovine granulosa cells is stimulated by insulin and insulin-like growth factor-I (IGF-I). Here we have assessed the effects of insulin and IGF-I on oxytocin gene expression in bovine granulosa cells and the dependence of these effects on the developmental status of the cells. When cells from individual follicles were cultured, the estradiol concentration of the follicular fluid was highly correlated with insulin stimulated oxytocin release. Subsequently, cells were pooled from follicles selected on the basis of estradiol content, and two subsets of cells were distinguished. The first contained highly differentiated cells, as judged by the high estradiol (HE-cells) concentration of the follicular fluid (greater than 40 ng/ml), high levels of LH receptors, and high hCG-stimulated cAMP accumulation. The second subset contained cells from follicles with low estradiol (less than 1 ng/ml; LE-cells) which have fewer LH receptors and low hCG-stimulated cAMP accumulation. Oxytocin production was increased more than 50-fold by insulin (EC50, 230 +/- 57 ng/ml) and IGF-I (EC50, greater than 10 ng/ml), but only in the HE-cells. Oxytocin mRNA was also greatly increased by insulin and IGF-I in the HE cells only. In contrast, insulin and IGF-I stimulated progesterone release from both HE- and LE-cells. Since oxytocin production is a characteristic of bovine luteal cells, our results support possible roles for IGF-I and insulin in regulation of luteinization or luteal activity. The data suggest that effects of insulin and IGF-I on oxytocin production reflect their effects on oxytocin gene transcription, and that granulosa cells must be appropriately primed (presumably by the in vivo hormonal environment) before they are able to produce oxytocin in response to these polypeptides. PMID- 2551660 TI - Protein kinase-C-induced down-regulation of calcitonin receptors and calcitonin activated adenylate cyclase in T47D and BEN cells. AB - T47D human breast cancer cells and BEN human lung cancer cells were preincubated with the tumor-promoting phorbol ester phorbol 12-myristate 13-acetate (PMA). In both cell lines there was a decrease in the binding of 125I-labeled salmon calcitonin ([125I]sCT) which was dependent on the dose and time of exposure to PMA. The effect on binding comprised at least two components: the apparent affinity for binding of [125I]sCT was decreased by PMA, and the rate of internalization of bound [125I]sCT was increased more than 2-fold in the presence of PMA. By using dinitrophenol to inhibit cellular metabolic energy and, therefore, receptor internalization, the PMA effects on receptor affinity were dissociated from those on endocytosis. The effects on binding were reflected in a decreased stimulation by sCT of adenylate cyclase activity. This was specific for the calcitonin receptor system, since PMA had no effect on prostaglandin-E2 stimulated adenylate cyclase in the T47D cell. Protein kinase-C (PKC) was implicated in the inhibitory effects of PMA on both binding and adenylate cyclase activation, since inhibition was reduced by simultaneous incubation with the PKC inhibitors H7 and H8. These results suggest that PKC is capable of mediating down regulation of the CT receptor, and this is most likely by phosphorylation of the receptor itself or an associated protein. PMID- 2551661 TI - Serotonin agonists cause parallel activation of the sympathoadrenomedullary system and the hypothalamo-pituitary-adrenocortical axis in conscious rats. AB - The effects of three potent serotonin (5-HT) agonists with different structures and 5-HT receptor binding profiles on sympathoadrenomedullary and hypothalamo pituitary-adrenocortical axis functions were assessed in conscious, freely moving male Sprague-Dawley rats. The 5-HT1A agonist, 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT), the 5-HT1C agonist, m-chlorophenylpiperazine (m CPP), and the 5-HT2/5-HT1C agonist, 1-(2,5-dimethoxy-4-iodophenyl)2-amino-propane (DOI), all produced marked dose-dependent increases in plasma epinephrine and ACTH concentrations. Both epinephrine and ACTH responses peaked at 10 min and showed strong positive correlations across all drugs and doses studied. Corticosterone increases showed a saturable response pattern and were close to the maximum level with a relatively small (approximately 2.5-fold) increase in plasma ACTH concentrations. Norepinephrine levels showed small dose-dependent increases after 8-OH-DPAT and m-CPP and decreases after DOI treatment. These results suggest that both the sympathoadrenomedullary system and the hypothalamo pituitary-adrenocortical axis can be activated via 5-HT1 and 5-HT2 receptors and that these two systems may have common or similar regulatory mechanisms triggered by these stimuli. PMID- 2551662 TI - Melatonin signal transduction in hamster brain: inhibition of adenylyl cyclase by a pertussis toxin-sensitive G protein. AB - Melatonin signal transduction was examined in median eminence/pars tuberalis (ME/PT) explants from Djungarian hamsters. High affinity melatonin receptors in hamster ME/PT were first quantified by in vitro autoradiography using the potent melatonin agonist 125I-labeled melatonin ([125I]MEL). Scatchard analysis of [125I]MEL binding in ME/PT revealed high affinity receptors [dissociation constant (Kd) = 2.75 X 10(-11) M]. [125I]MEL binding was markedly reduced by guanine nucleotides; treatment with the nonhydrolyzable GTP analog guanosine 5'-O (3-thiotriphosphate) caused a 10-fold decrease in receptor affinity. Melatonin (10 nM) significantly inhibited forskolin-stimulated cAMP accumulation in ME/PT, but not in pituitary or pineal glands. In ME/PT explants, melatonin and 6 chloromelatonin inhibited forskolin-stimulated cAMP accumulation in a dose dependent manner with similar potency (significant inhibition for each at concentrations greater than or equal to 100 pM). Serotonin significantly inhibited forskolin-stimulated cAMP levels only at doses greater than or equal to 100 microM. Inhibition of [125I]MEL binding in ME/PT by these three indolamines paralleled that determined for inhibition of forskolin-stimulated cAMP accumulation. Pertussis toxin treatment (1 microgram/ml) blocked the ability of melatonin (10 nM) to inhibit forskolin-stimulated cAMP accumulation and significantly reduced [125I]MEL binding. Pertussis toxin ADP-ribosylated the alpha-subunits of at least two guanine nucleotide-binding proteins in ME/PT explants with molecular weights of approximately 40 K. Melatonin did not increase phosphodiesterase activity in ME/PT explants. The results strongly suggest that a signal transduction pathway for melatonin in mammals involves inhibition of adenylyl cyclase by a pertussis toxin-sensitive guanine nucleotide-binding protein. PMID- 2551663 TI - Laparoscopic observation of hepatic lobe atrophy. AB - From the standpoint of diagnostic laparoscopy, the frequency and etiological aspect of atrophy of a lobe of the liver was studied. The frequency of hepatic lobe atrophy was 5.3% among 1,208 laparoscopy cases at our department. Lobe atrophy of the liver can be seen not only in congenital anomaly, atrophic cirrhosis and malformation, as described in the OMED database of digestive endoscopy, but also in some other kinds of liver diseases including chronic hepatitis, idiopathic portal hypertension, primary biliary cirrhosis, drug induced liver injury, scarred liver, autoimmune hepatitis and also in malignancies of other visceral organs. The disorders most frequently associated with hepatic lobe atrophy were idiopathic portal hypertension, and scarred liver, primary biliary cirrhosis, etc. PMID- 2551664 TI - Metabolism of phenol and hydroquinone to reactive products by macrophage peroxidase or purified prostaglandin H synthase. AB - Macrophages, an important cell-type of the bone marrow stroma, are possible targets of benzene toxicity because they contain relatively large amounts of prostaglandin H synthase (PHS), which is capable of metabolizing phenolic compounds to reactive species. PHS also catalyzes the production of prostaglandins, negative regulators of myelopoiesis. Studies indicate that the phenolic metabolites of benzene are oxidized in bone marrow to reactive products via peroxidases. With respect to macrophages, PHS peroxidase is implicated, as in vivo benzene-induced myelotoxicity is prevented by low doses of nonsteroidal anti inflammatory agents, drugs that inhibit PHS. Incubations of either 14C-phenol or 14C-hydroquinone with a lysate of macrophages collected from mouse peritoneum (greater than 95% macrophages), resulted in an irreversible binding to protein that was dependent upon H2O2, incubation time, and concentration of radiolabel. Production of protein-bound metabolites from phenol or hydroquinone was inhibited by the peroxidase inhibitor aminotriazole. Protein binding from 14C-phenol also was inhibited by 8 microM hydroquinone, whereas binding from 14C-hydroquinone was stimulated by 5 mM phenol. The nucleophile cysteine inhibited protein binding of both phenol and hydroquinone and increased the formation of radiolabeled water soluble metabolites. Similar to the macrophage lysate, purified PHS also catalyzed the conversion of phenol to metabolites that bound to protein and DNA; this activation was both H2O2- and arachidonic acid-dependent. These results indicate a role for macrophage peroxidase, possibly PHS peroxidase, in the conversion of phenol and hydroquinone to reactive metabolites and suggest that the macrophage should be considered when assessing the hematopoietic toxicity of benzene. PMID- 2551665 TI - Peroxidase-dependent metabolism of benzene's phenolic metabolites and its potential role in benzene toxicity and carcinogenicity. AB - The metabolism of two of benzene's phenolic metabolites, phenol and hydroquinone, by peroxidase enzymes has been studied in detail. Studies employing horseradish peroxidase and human myeloperoxidase have shown that in the presence of hydrogen peroxide phenol is converted to 4,4'-diphenoquinone and other covalent binding metabolites, whereas hydroquinone is converted solely to 1,4-benzoquinone. Surprisingly, phenol stimulates the latter conversion rather than inhibiting it, an effect that may play a role in the in vivo myelotoxicity of benzene. Indeed, repeated coadministration of phenol and hydroquinone to B6C3F1 mice results in a dramatic and significant decrease in bone marrow cellularity similar to that observed following benzene exposure. A mechanism of benzene-induced myelotoxicity is therefore proposed in which the accumulation and interaction of phenol and hydroquinone in the bone marrow and the peroxidase-dependent formation of 1,4 benzoquinone are important components. This mechanism may also be responsible, at least in part, for benzene's genotoxic effects, as 1,4-benzoquinone has been shown to damage DNA and is shown here to induce multiple micronuclei in human lymphocytes. Secondary activation of benzene's phenol metabolites in the bone marrow may therefore play an important role in benzene's myelotoxic and carcinogenic effects. PMID- 2551666 TI - Multifunctional receptor model for dioxin and related compound toxic action: possible thyroid hormone-responsive effector-linked site. AB - Molecular/theoretical modeling studies have revealed that thyroid hormones and toxic chlorinated aromatic hydrocarbons of environmental significance (for which dioxin or TCDD is the prototype) have similar structural properties that could be important in molecular recognition in biochemical systems. These molecular properties include a somewhat rigid, sterically accessible and polarizable aromatic ring and size-limited, hydrophobic lateral substituents, usually contained in opposite adjoining rings of a diphenyl compound. These molecular properties define the primary binding groups thought to be important in molecular recognition of both types of structures in biochemical systems. Similar molecular reactivities are supported by the demonstration of effective specific binding of thyroid hormones and chlorinated aromatic hydrocarbons with four different proteins, enzymes, or receptor preparations that are known or suspected to be involved in the expression of thyroid hormone activity. These binding interactions represent both aromatic-aromatic (stacking) and molecular cleft-type recognition processes. A multiple protein or multifunctional receptor-ligand binding mechanism model is proposed as a way of visualizing the details and possible role of both the stacking and cleft type molecular recognition factors in the expression of biological activity. The model suggests a means by which hormone-responsive effector-linked sites (possible protein-protein-DNA complexes) can maintain highly structurally specific control of hormone action. Finally, the model also provides a theoretical basis for the design and conduct of further biological experimentation on the molecular mechanism(s) of action of toxic chlorinated aromatic hydrocarbons and thyroid hormones. PMID- 2551667 TI - Mechanisms of free radical chemistry and biochemistry of benzene. AB - omicron-Tyrosine (omicron-Tyr) was used as a specific biomarker for .OH radicals generated in biosystems. Specificity of omicron-Tyr as an .OH biomarker was based on previous studies in systems exposed to ionizing radiations. Fresh muscle tissue incubated with benzene for 1 hr at 38 degrees C exhibits formation of omicron-Tyr as seen in the cases of ethanol- and carbon tetrachloride-exposed systems. Gas chromatography/mass spectrometry selective ion monitoring measurements of omicron-Tyr yields in chicken breast muscle incubated with water or benzene indicate levels of less than 0.1 ppm and 3.0 +/- 0.5 ppm of omicron Tyr, respectively. Formation of .OH is presumed to originate via a Haber-Weiss reaction of H2O2 with Fe(II) preceded by the formation of .O2- and H2O2 from distorted mitochondria. PMID- 2551669 TI - Expression of normal and translocated c-myc alleles in Burkitt's lymphoma cells: evidence for different regulation. AB - In Burkitt's lymphoma (BL) cells the normal c-myc allele is usually silent or expressed at very low levels. Here we demonstrate that the normal c-myc allele can be induced in BL cells by 12-O-tetradecanoylphorbol-13-acetate (TPA). TPA did activate the normal c-myc alleles in Raji(P207), BL36, P3HR1, Jijoye and LY91 cells, but not in Raji(DE88), BL41, BL67, LY47 and KK124 cells. C-myc RNA derived from the normal allele appeared 6 h after treatment with TPA and showed the characteristic preferential usage of the second promoter. This induction could not be inhibited by cycloheximide. Despite the differences in c-myc induction in Raji(P207) and Raji(DE88) cells, c-fos and the early Epstein-Barr virus gene DR were induced to a similar extent and with similar kinetics by TPA. Nuclear run-on experiments suggest that the normal c-myc allele in Raji cells is activated at least in part by releasing a block to RNA elongation at the end of c-myc exon 1. Expression of the translocated c-myc alleles was also affected by TPA; however, only if cycloheximide was simultaneously present. TPA plus cycloheximide induced a rapid decrease of c-myc RNA derived from the translocated allele within 6 h, whereas cycloheximide alone led to abolition of c-myc RNA after 16-24 h. This rapid decline of c-myc RNA was observed in Raji and BL41 cells, but not in three cell lines with variant t(2;8) and t(8;22) translocations. PMID- 2551668 TI - Sequence analysis of alpha 1(VI) and alpha 2(VI) chains of human type VI collagen reveals internal triplication of globular domains similar to the A domains of von Willebrand factor and two alpha 2(VI) chain variants that differ in the carboxy terminus. AB - Amino acid sequences of human collagen alpha 1(VI) and alpha 2(VI) chains were completed by cDNA sequencing and Edman degradation demonstrating that the mature polypeptides contain 1009 and 998 amino acid residues respectively. In addition, they contain small signal peptide sequences. Both chains show 31% identity in the N-terminal (approximately 235 residues) and C-terminal (approximately 430 residues) globular domains which are connected by a triple helical segment (335 336 residues). Internal alignment of the globular sequences indicates a repetitive 200-residue structure (15-23% identity) occurring three times (N1, C1, C2) in each chain. These repeating subdomains are connected to each other and to the triple helix by short (15-30 residues) cysteine-rich segments. The globular domains possess several N-glycosylation sites but no cell-binding RGD sequences, which are exclusively found in the triple helical segment. Sequencing of alpha 2(VI) cDNA clones revealed two variant chains with a distinct C2 subdomain and 3' non-coding region. The repetitive segments C1, C2 and, to a lesser extent, N1 show significant identity (15-18%) to the collagen-binding A domains of von Willebrand factor (vWF) and they are also similar to some integrin receptors, complement components and a cartilage matrix protein. Since the globular domains of collagen VI come into close contact with triple helical segments during the formation of tissue microfibrils it suggests that the globular domains bind to collagenous structures in a manner similar to the binding of vWF to collagen I. PMID- 2551670 TI - Post-transcriptional mechanisms of deregulation of MYC following conversion of a human B cell line by Epstein-Barr virus. AB - By utilizing an Epstein-Barr virus (EBV)-negative Burkitt's lymphoma line (BJAB) and several EBV-positive sub-lines derived from it by in vitro infection, it has been shown previously that the presence of the EBV genome in this B cell line is reversibly associated with deregulation of MYC expression. Specifically, the decline in the level of MYC transcripts observed in the EBV-negative BJAB line as cells approach stationary phase of growth is abrogated in the presence of the virus. In the studies described herein, the mechanism of deregulation of MYC in EBV-converted BJAB cells was examined. It was shown that the presence of EBV in BJAB cells was not associated with changes in the rate of transcription from exons I, II or III of MYC as cells approached stationary phase of growth. In contrast, the stability of MYC mRNA was altered in EBV-positive BJAB cells. Specifically, the half-life of MYC mRNA increased from less than 36 to greater than 70 min in EBV-positive BJAB lines as cells progressed from early to late exponential phase of growth. This alteration in stability of MYC transcripts was reversibly associated with the presence of the virus, since an EBV-negative revertant BJAB line did not display an increase in stability of MYC transcripts in late exponential phase of growth. In addition, progression from early to late exponential phase of growth in two EBV-immortalized lymphoblastoid lines derived from normal human B cells was also associated with prolongation of the half-life of MYC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551671 TI - Insulin-like growth factor 2 and short-range stimulatory loops in control of human placental growth. AB - Substructures of the first-trimester human placenta (within 3 months post conception) display 'pseudo-malignant' properties. We show here, by in situ hybridization, that the insulin-like growth factor 2 (IGF-2) gene expression is particularly active in the cytotrophoblasts, which dominate these structures. Because the majority of placental IGF-2 mRNA is polysomal in extracts of first trimester placenta, the spatial pattern of IGF-2 transcripts generally also defines the pattern of IGF-2 production. In primary trophoblast cultures, rendered quiescent by serum starvation. IGF-2 performs as a human embryonic growth factor by activating cell cycle entry/progression. Although both type 1 and 2 IGF receptor mRNAs can be found co-distributed with IGF-2 mRNA during placental development (supporting an autocrine role for IGF-2), these occasional patterns are confined to cytotrophoblasts with low proliferative potential. The reciprocity in ligand and receptor expression patterns are discussed in terms of rate-limiting steps in the involvement of IGF-2 in the proliferative phenotype of the early human placenta. PMID- 2551672 TI - The unique insert of cellular and viral fms protein tyrosine kinase domains is dispensable for enzymatic and transforming activities. AB - The receptors for colony stimulating factor-1 (CSF-1), platelet derived growth factor and the c-kit protein tyrosine kinase (PTK) contain within their catalytic domains a stretch of 60-100 residues, largely unrelated in sequence, with no counterpart in other PTKs. Of the 64 amino acids within this kinase insert, 58 were deleted from the mouse CSF-1 receptor by oligonucleotide-directed mutagenesis. The mutant CSF-1 receptor was not markedly affected in its kinase activity, post-translational processing or its ability to induce autocrine transformation of NIH 3T3 mouse fibroblasts. Similarly, retention of kinase and transforming activities were observed following deletion of part or all of the kinase insert from the v-fms oncoprotein. The c- and v-fms kinase inserts were probed using monoclonal and polyclonal antibodies and were found to be highly antigenic. Two monoclonal antibodies raised to the v-fms cytoplasmic domain both recognized epitopes within the insert, and bound enzymatically active v-fms glycoproteins. These results indicate that the fms kinase insert is located on the surface of the protein and folds separately from the rest of the catalytic domain, but is not required for the biological activity of fms PTKs ectopically expressed in mouse fibroblasts. The insert may therefore play a specific function in cells such as monocytes and trophoblasts that normally express the CSF-1 receptor. PMID- 2551673 TI - Overproduction of the regulatory subunit of the cAMP-dependent protein kinase blocks the differentiation of Dictyostelium discoideum. AB - During the aggregation of Dictyostelium discoideum extracellular cAMP is known to act as a chemotractant and as an inducer of cellular differentiation. However, its intracellular role as a second messenger remains obscure. We have constructed a fusion gene consisting of the cDNA encoding the regulatory subunit (R) of the cAMP-dependent protein kinase fused to the promoter and N-terminal-proximal sequences of a Dictyostelium actin gene. Stable transformants, containing multiple copies of this gene, overproduce the R subunit which accumulates prematurely relative to the endogenous protein. These transformants fail to aggregate. Detailed analysis has shown that they are blocked at interphase, the period prior to aggregation, and that they are severely defective in most responses to cAMP including the induction of gene expression. Our observations suggest that intracellular cAMP acts, presumably by activation of the catalytic subunit of the cAMP-dependent protein kinase, to facilitate early development. PMID- 2551674 TI - Identification and characterization of genes and mutants for an N-terminal acetyltransferase from yeast. AB - A gene from Saccharomyces cerevisiae has been mapped, cloned, sequenced and shown to encode a catalytic subunit of an N-terminal acetyltransferase. Regions of this gene, NAT1, and the chloramphenicol acetyltransferase genes of bacteria have limited but significant homology. A nat1 null mutant is viable but exhibits a variety of phenotypes, including reduced acetyltransferase activity, derepression of a silent mating type locus (HML) and failure to enter G0. All these phenotypes are identical to those of a previously characterized mutant, ard1. NAT1 and ARD1 are distinct genes that encode proteins with no obvious similarity. Concomitant overexpression of both NAT1 and ARD1 in yeast causes a 20-fold increase in acetyltransferase activity in vitro, whereas overexpression of either NAT1 or ARD1 alone does not raise activity over basal levels. A functional iso-1 cytochrome c protein, which is N-terminally acetylated in a NAT1 strain, is not acetylated in an isogenic nat1 mutant. At least 20 other yeast proteins, including histone H2B, are not N-terminally acetylated in either nat1 or ard1 mutants. These results suggest that NAT1 and ARD1 proteins function together to catalyze the N-terminal acetylation of a subset of yeast proteins. PMID- 2551675 TI - Mutational analysis of IS10's outside end. AB - We present the genetic analysis of a large number of mutations in the outside end of insertion sequence IS10. (i) The terminal inverted repeat sequence is probably the primary site of transposase binding. Mutations in this region fall into phenotypic classes which correspond to their map locations, suggesting that this region may consist of several distinct functional segments. Similarities between the organization of IS10's inverted repeat and those of other transposable elements are discussed. (ii) Base pairs 23-42 include a consensus binding sequence for one of the IS10 transposition host factors, IHF. The phenotypes of mutations in this region suggest that IHF is the major host factor for outside end transposition activity in vivo and that base pairs throughout this region are important for the IHF interaction. (iii) Mutations in bp 43-61 do not affect outside-end transposition activity but do affect, in expected ways, previously identified determinants involved in expression and regulation of transposase. (iv) Some mutations in bp 23-42 also affect transposase expression; the possibility that IHF negatively regulates transcription initiation is discussed. PMID- 2551676 TI - Expression of MyoD1 coincides with terminal differentiation in determined but inducible muscle cells. AB - We have examined the expression of MyoD1, a potential determination factor of myogenic cells, in permissive and inducible C2 myoblasts. These two types of myoblasts exhibit distinct requirements to undergo terminal differentiation. Unlike permissive cells, inducible cells fail to differentiate in the presence of growth medium plus fetal calf serum and require insulin to undergo terminal differentiation. We show that while expression of MyoD1 is constitutive in permissive cells, no trace of MyoD1 transcripts is found in inducible cells at the myoblast stage. In these cells, however, expression of MyoD1 accompanies differentiation. This indicates that MyoD1 may not be required for the maintenance of the myoblast phenotype, and could act as an effector of terminal differentiation in already determined muscle cells. Our results provide new evidence that permissive and inducible cells represent two distinct stages of the progression of determined muscle cells toward terminal differentiation. PMID- 2551677 TI - A membrane component of the endoplasmic reticulum that may be essential for protein translocation. AB - We have purified a glycosylated, membrane-spanning protein of relative molecular mass approximately 34,000 (Mr approximately 34 K) from canine microsomes that appears to be essential for protein translocation across the endoplasmic reticulum (ER) as shown by the inhibitory action of antibodies directed against it and of monovalent Fab-fragments produced from them. The ER membrane contains at least as many molecules of the 34 K membrane protein as bound ribosomes. The protein can be detected immunologically in tissues of various organisms, indicating an universal function. PMID- 2551678 TI - Fibroblast growth factor phosphorylation and receptors in rod outer segments. AB - Acidic and basic fibroblast growth factors (aFGF and bFGF) have been isolated and purified from rod outer segments (ROS). aFGF is tightly bound to ROS membranes and can be specifically released by ATP. We show that this mechanism is dependent on the phosphorylation of aFGF itself. Phorbol 12-myristate 13-acetate (PMA) enhances this phenomenon independently of rhodopsin phosphorylation. This demonstrates that aFGF release from ROS membranes is dependent on its phosphorylation by endogenous kinase C. In addition specific binding sites for exogenous FGFs have been identified on ROS and disc membranes. A single high affinity site with a Kd of 40 pM was present in intact ROS while an additional low affinity site with a Kd of 300-600 pM was present in leaky ROS or in disc membranes. Light or ATP modified neither these Kd nor the apparent number of sites. The presence of specific receptors for FGFs and the kinase C dependent release of endogenous membrane bound aFGF suggest an autocrine mechanism which may be involved in photoreceptor cell biology. PMID- 2551680 TI - The interference of truncated with normal potassium channel subunits leads to abnormal behaviour in transgenic Drosophila melanogaster. AB - The Shaker locus of Drosophila melanogaster encodes a family of A-type potassium channel subunits. Shaker mutants behave as antimorphs in gene dosage tests. This behaviour is due to the production of truncated A-channel subunits. We propose that they interfere with the function of their normal counterpart by forming multimeric A-channel structures. This hypothesis was tested by constructing transgenic flies carrying a heat-inducible gene encoding a truncated A-type potassium channel subunit together with a normal wild type doses of A-type potassium channel subunits. The altered subunit leads at larval, pupal or adult stages to the transformation of wild type into Shaker flies. The transformed flies exhibited a heat-inducible abnormal leg shaking behaviour and a heat inducible facilitated neurotransmitter release at larval neuromuscular junctions. By the overexpression of an aberrant A-channel subunit the normal behaviour of transgenic D. melanogaster can be altered in a predictable way. PMID- 2551679 TI - Cyclin is a component of the sea urchin egg M-phase specific histone H1 kinase. AB - A so-called 'growth-associated' or 'M-phase specific' histone H1 kinase (H1K) has been described in a wide variety of eukaryotic cell types; p34cdc2 has previously been shown to be a catalytic subunit of this protein kinase. In fertilized sea urchin eggs the activity of H1K oscillates during the cell division cycle and there is a striking temporal correlation between H1K activation and the accumulation of a phosphorylated form of cyclin. H1K activity declines in parallel with proteolytic cyclin destruction of the end of the first cell cycle. By virtue of the high affinity of the fission yeast p13suc1 for the p34cdc2 protein, H1K strongly binds to p13-Sepharose beads. Cyclin, p34cdc2 and H1K co purify on this affinity reagent as well as through several conventional chromatographic procedures. Anticyclin antibodies immunoprecipitate the M-phase specific H1K in crude extracts or in purified fractions. Sea urchin eggs appear to contain much less cyclin than p34cdc2, suggesting that p34cdc2 may interact with other proteins. These results demonstrate that cyclin and p34cdc2 are major components of the M-phase specific H1K. PMID- 2551681 TI - Diversity and novel pharmacological properties of Ca2+ channels in Drosophila brain membranes. AB - Binding studies as well as affinity labelling and immunoblot techniques were used to identify and characterize the receptors for Ca2+ channel blockers in Drosophila brain membranes. Despite structural analogies with mammalian receptors, Drosophila binding sites for phenylalkylamines and 1,4 dihydropyridines, unlike those described in skeletal and cardiac muscle, were found to be located on separate Ca2+ channels. Single-channel bilayer recordings from reconstituted membranes revealed the presence of eight distinct cobalt sensitive Ba2+-conducting channels in Drosophila brain membrane preparations. In good agreement with binding studies, the most frequently observed Ca2+ channel type (Ba2+ conductance of 13 pS) was extremely sensitive to phenylalkylamines but not affected by micromolar concentrations of 1,4-dihydropyridines. Distinct 1,4 dihydropyridine-sensitive and phenylalkylamine-insensitive channels were also identified. They had unitary Ba2+ conductances of 21 and 31 pS. A detailed analysis of drug action showed that both 1,4-dihydropyridines and phenylalkylamines first increased channel open state probability before fully blocking channel activity. Other types of channels have been identified with unitary Ba2+ conductances of 9, 41, 53, 64 and 81 pS. They were insensitive to the previously described organic Ca2+ channel blockers. The Drosophila system seems to be a unique model to analyse the properties of several different types of Ca2+ channels and particularly those of channel types that are uniquely blocked by phenylalkylamines or uniquely blocked by 1,4-dihydropyridines. PMID- 2551682 TI - The Escherichia coli regulatory protein OxyR discriminates between methylated and unmethylated states of the phage Mu mom promoter. AB - Expression of the phage Mu mom gene is transcriptionally regulated by DNA methylation. Three GATC sites upstream of the mom promoter have to be methylated by the Escherichia coli deoxyadenosine methylase (Dam) to allow initiation of transcription. An E. coli dam strain was mutagenized with Tn5 in an attempt to isolate mutants which allow mom gene expression. Three independent Tn5 mutants were isolated, each mapped to a gene at 89.6 min which we designate momR. The wildtype gene was cloned and sequenced, it encodes a protein of 305 amino acids. The protein belongs to a group of related bacterial activators recently identified as the LysR family (Henikoff et al., 1988). MomR protein was overproduced and purified. Expression of momR is autoregulated; MomR binds to a 43 bp region upstream of its coding sequence. In the mom promoter MomR protects a 43 bp region containing the three GATC sites. Specific binding to these sequences was observed only with unmethylated DNA. Fortuitously, we learned that MomR is identical to OxyR, a regulatory protein responding to oxidative stress. We discuss the implications of this control for Mu development. PMID- 2551685 TI - Serological analysis and characterization of calf thymus ribonuclease H IIb. AB - Ribonuclease H IIb, which seems to play a physiological role during transcription, was purified from calf thymus tissue. A polyclonal antibody, raised against the most purified ribonuclease H IIb fraction, recognizes in crude extracts almost exclusively a 52-kDa protein band. By immunoaffinity chromatography and immunoprecipitation experiments, we are able to deplete enzyme extracts from the crossreacting 52-kDa protein band and from ribonuclease H IIb activity. Enzyme activity is eluted from the immunoaffinity matrix in association with a 52-kDa protein under denaturing conditions. Immunoaffinity chromatography enables us also to calculate a purification factor of around 20,000 from the crude extract. The native molecular mass for the enzyme of around 45 kDa, as determined by gel filtration, suggests that calf thymus ribonuclease H IIb is most probably monomeric. The enzyme possesses an isoelectric point of 7.0. It requires Mg2+ ions for activity, is inhibited by N-ethylmaleimide, and exhibits a pH optimum of 9.0-9.5. The enzyme releases oligoribonucleotides with 3'-OH and 5' phosphate ends, probably in an exonucleolytical manner. The third largest subunit of yeast RNA polymerase A (I) displays ribonuclease H activity [Huet et al. (1976) Nature 261, 431-433]. We discuss our findings in the light of a possible association of ribonuclease H IIb and RNA polymerase A (I) in higher eukaryotes. PMID- 2551684 TI - Evidence of a ter specific binding protein essential for the termination reaction of DNA replication in Escherichia coli. AB - Activity binding specifically to the 22 bp of the DNA replication terminus (ter) sequence on plasmid R6K and the Escherichia coli genome was detected in the crude extract of E. coli cells. This activity was inactivated by heat or by protease but not by RNase treatments. Overproduction of the ter binding activity was observed when the extract was prepared from the cell carrying a plasmid with a chromosomal-derived 5.0 kb EcoRI fragment, on which one of the four terC sites, terC2, was also located. By mutagenesis of the 5.0 kb fragment on the plasmid with transposon Tn3 and subsequent replacement of the corresponding chromosomal region with the resulting mutant alleles, we isolated tau- mutants completely defective in ter binding activity. These mutants simultaneously lost the activity to block the progress of the DNA replication fork at any ter site, on the genome or the plasmid. It would thus appear that the ter binding protein plays an essential role in the termination reaction, at the ter sites. PMID- 2551683 TI - Molecular characterization of two proteins involved in the excision of the conjugative transposon Tn1545: homologies with other site-specific recombinases. AB - Excision is probably the initial and rate-limiting step of the movements of conjugative transposons of Gram-positive bacteria such as Tn916 and Tn1545. We have shown, by molecular cloning and DNA sequencing, that a 2058 bp Sau3A right junction fragment of transposon Tn1545 specifies two gene products that are involved in the excision of the element. The DNA sequence of these genes, designated orf1 and orf2, has been determined and the corresponding proteins, ORF1 and ORF2, have been identified in a bacterial cell-free coupled transcription-translation system. These proteins are freely diffusible since they are able to trans-complement in vivo a deletion derivative of Tn1545 defective for excision. Using an in vivo complementation assay, we have demonstrated that ORF2 alone is able to catalyse excision and that ORF1 strongly stimulates the activity of ORF2. We also found that ORF1 and ORF2 display local homology with, respectively, proteins Xis and Int from lamboid phages, which suggests that these excision systems have a common origin. Based on the functional properties of the integrase of bacteriophage lambda, on the analysis of the nucleotide sequence of the junction fragments and of the target before insertion and after excision, a model is proposed for ORF2-catalysed excision of Tn1545 and related conjugative transposons. PMID- 2551687 TI - Presence of fucosylated triantennary, tetraantennary and pentaantennary glycans in transferrin synthesized by the human hepatocarcinoma cell line Hep G2. AB - Transferrin synthesized by a human hepatocellular carcinoma cell line Hep G2 (called Hep G2 transferrin) was purified from culture media by immunoaffinity chromatography on a rabbit anti-(human serotransferrin) IgG column. The eluted transferrin was then resolved into five peaks on a cation-exchange column using the fast protein liquid chromatography system. The major fraction, named Hep G2 transferrin fraction C, having a molecular mass of 82.5 kDa was found to be homogeneous in polyacrylamide gel electrophoresis and in concanavalin-A-affinity crossed immunoelectrophoresis. A comparative analysis of the molar carbohydrate composition of normal human serotransferrin and of Hep G2 transferrin fraction C shows an increase in the latter in the number of galactose and N acetylglucosamine residues and in the presence of fucose, which is absent in normal transferrin. By a combination of methylation analysis and NMR spectroscopy, the primary structure of the oligosaccharide alditols released from Hep G2 transferrin fraction C by reductive alkaline cleavage has been established as triantennary, tetraantennary and pentaantennary N-acetyllactosaminic structures with fucose residues (alpha 1-3)-linked to peripheral N acetylglucosamine residues. These results indicate that the increase in the number of antennae in transferrin glycans synthesized by the hepatocarcinoma cell line is much more pronounced than in liver diseases such as alcoholic cirrhosis and that, in addition, the malignant transformation of human liver induces the presence of fucose. PMID- 2551686 TI - Synthesis of a gene for the protein kinase domain of the epidermal growth factor receptor and its expression in Escherichia coli. AB - A gene encoding the protein kinase domain of the epidermal growth factor receptor has been chemically synthesised, cloned and expressed in Escherichia coli. The 942-base-pair gene was constructed by enzymatic ligation of 56 oligonucleotides and cloned into an expression vector downstream of the E. coli trp promoter. Production of active gene product was confirmed by means of a protein kinase assay, demonstrating that the enzymatic activity of the protein kinase domain of the epidermal growth factor receptor is retained after expression in E. coli. PMID- 2551688 TI - Guanylate kinase from Saccharomyces cerevisiae. Isolation and characterization, crystallization and preliminary X-ray analysis, amino acid sequence and comparison with adenylate kinases. AB - This paper describes a large-scale purification of guanylate kinase (ATP + GMP in equilibrium ADP + GDP) from Saccharomyces cerevisiae, the crystallization of the enzyme and preliminary X-ray investigations. Furthermore the complete amino acid sequence of the enzyme has been determined and was compared to adenylate kinase sequences. 1. Guanylate kinase was purified in five steps to homogeneity: crude extract, ion-exchange chromatography, affinity chromatography and gel filtration twice. 2. The enzyme was crystallized to single octahedral bipyramids with sizes up to 500 x 200 x 150 microns 3. Preliminary X-ray results are given. 3. The final sequence shows 186 amino acids (Mr = 20,548), containing one cysteine and one tryptophan. It was determined from peptides of five cleavages of the whole protein. Three cleavages were used for determination of the whole polypeptide chain. From the other two, only some peptides were used to secure overlaps and the cysteine position. The N-terminal blocking group was identified by 1H-NMR spectroscopy. 4. Since guanylate kinase shows the mononucleotide binding pattern GXXGXGK, it was compared to other proteins containing this pattern. But no further homology signal could be detected. A comparison with adenylate kinases revealed significant similarity in another chain segment. This led to the conclusion that guanylate kinase is at least partially homologous to the adenylate kinases. PMID- 2551689 TI - Synthesis of different nucleoside 5'-diphospho-sulfoquinovoses and their use for studies on sulfolipid biosynthesis in chloroplasts. AB - 6-Sulfo-alpha-D-quinovopyranosyl phosphate was reacted with different nucleoside monophosphate morpholidates to form ADP-, CDP-, GDP- and UDP-sulfoquinovose. Analytical and preparative HPLC of these nucleotides was performed on reversed phase columns using volatile buffer systems as eluant. The isolated compounds were characterized by NMR spectroscopy (except the CDP derivative) and used for an investigation of sulfolipid biosynthesis by chloroplasts. For this purpose intact spinach chloroplasts were biosynthetically preloaded with radioactive diacylglycerol to provide a sulfoquinovosyl acceptor. When sulfosugar nucleotides were added to such prelabelled intact organelles, the background levels of sulfolipid biosynthesis did not rise. On the other hand, after osmotic shock of prelabelled chloroplasts sulfolipid labelling was significantly increased by the addition of UDP- or GDP-sulfoquinovose. The same stimulation was observed with isolated envelope membranes, and UDP-sulfoquinovose proved to be twice as active as the GDP derivative. From these results it was concluded that the final step in sulfolipid biosynthesis is catalyzed by a UDP-sulfoquinovose: 1,2-diacylglycerol 3-O-alpha-D-sulfoquinovosyltransferase. This chloroplast enzyme cannot use exogenously supplied sulfosugar nucleotides, which as membrane-impermeable compounds are expected to be formed in vivo within chloroplasts. PMID- 2551690 TI - Characterization of a mitogen-activated, Ca2+-sensitive microtubule-associated protein-2 kinase. AB - We have previously found that treatment of quiescent mammalian fibroblast cells with several mitogenic factors activates in common a Ca2+-sensitive serine/threonine-specific protein kinase activity toward microtubule-associated protein 2 (MAP2) [Hoshi, M., Nishida, E. and Sakai, H. (1988) J. Biol. Chem. 263, 5396-5401]. Here, we characterized the mitogen-activated MAP2 kinase activity in rat 3Y1 cells. The activated kinase activity was detected in the cytosolic fraction but not in the membrane fraction. The inhibitory effect of Ca2+ on the kinase activity was reversible. Kinetic analyses revealed that the apparent Km values of the kinase activity for MAP2 and ATP were 1.6 microM and 30 microM, respectively. Free Ca2+ at 4 microM decreased apparent Vmax values for MAP2 and ATP without changing the apparent Km values. The MAP2 kinase had an apparent molecular mass of about 40 kDa as determined by gel filtration and by sucrose density gradient centrifugation. Myelin basic protein as well as MAP2 could serve as good substrates for this kinase, but 40S ribosomal protein S6, casein, histone, phosphorylase b, protamine, tubulin, actin and tau could not. These properties of the enzyme indicate that the Ca2+-sensitive MAP2 kinase may be a previously unidentified enzyme. Down-regulation of protein kinase C by prolonged phorbol ester treatment abolished the MAP2 kinase activation by phorbol ester, but did not prevent the MAP2 kinase activation by epidermal growth factor (EGF) or fresh serum. This suggests that the Ca2+-sensitive MAP2 kinase could be activated through protein-kinase-C-dependent and -independent pathways. Activation of the MAP2 kinase occurred shortly after the addition of EGF or phorbol ester even in the presence of protein synthesis inhibitors (cycloheximide, puromycin and emetin). Moreover, treatment of the EGF- or phorbol ester-activated MAP2 kinase with acid phosphatase inactivated the kinase activity. Thus, the MAP2 kinase may be activated through phosphorylation. PMID- 2551692 TI - Hormonal background of the hypertension and fluid derangements associated with adrenocorticotrophic hormone treatment of infants. AB - We studied the hormonal background of the fluid derangements and arterial hypertension associated with adrenocorticotrophic hormone (ACTH) treatment for infantile spasms in ten infants aged 5-22 months. They received a 6 week course of (carboxymethyl-cellulose-)ACTH: 80 IU at 0800 hours daily in weeks 1-3, then tapering, and termination at the end of week 6. The infants showed large, variable increases in 24 h urine cortisol during treatment. The mean plasma cortisol concentration (24 h after ACTH injection) was not significantly increased, but was correlated with the relative dose of ACTH. The mean plasma aldosterone concentration decreased. No significant change occurred in plasma renin activity (PRA), or in the concentrations of renin substrate (RS) or arginine vasopressin (AVP). Seven infants developed arterial hypertension, which was severe in three. This severe hypertension was associated with the highest relative ACTH doses and the highest plasma RS and cortisol concentrations. In the group as a whole, systolic blood pressure correlated with plasma RS and cortisol concentrations, but not with the other parameters. At the end of treatment urine and plasma cortisol dropped below the pretreatment levels and stayed low for greater than 2 weeks. There was a sharp peak in PRA and plasma aldosterone concentration, and a decrease in plasma RS. Plasma AVP levels dropped markedly. The mean body weight increased sharply and urine flow decreased. Mean plasma electrolyte levels remained unaltered. The danger at termination of ACTH treatment appears to be associated with a sudden transition from hypercortisolism to hypocortisolism, activation of the renin-angiotensin-aldosterone axis, and suppression of AVP secretion. PMID- 2551693 TI - Rickets with alopecia-remission following a course of 1-alpha-hydroxy vitamin D3 therapy. AB - A child is described with rickets and alopecia who did not respond to high doses of vitamin D3 but who responded to a small dose of 1-alpha-hydroxyvitamin D3. Treatment was continued for 2 years and then stopped. She has not shown any signs of relapse 1 year after stopping treatment. Her alopecia, however, has remained unchanged. One year after stopping treatment, her serum 25-hydroxycholecalciferol and parathormone levels were within normal limits but serum 1,25 dihydroxycholecalciferol was elevated. PMID- 2551691 TI - Abdominal, retroperitoneal and sacrococcygeal tumours of the newborn and the very young infant. Report from the Kiel Paediatric Tumour Registry. AB - We examined 226 abdominal, retroperitoneal and sacrococcygeal tumours in newborns and infants aged 6 months or less. Most frequent were neuroblastomas (n = 83) followed by germ cell tumours (n = 76), 37 of which were immature and 32 were mature teratomas. Fully malignant germ cell tumours and malignant germ cell tumour components were much rarer in this age group (9.2%) than in older children (58.1%). The majority of germ cell tumours were localized in the sacrococcygeal region (72.4%). Next in frequency were tumours of the kidney (n = 54), including 28 congenital mesoblastic nephromas. In contrast to children over 6 months of age, kidney tumours were frequently low grade malignant with a favourable prognosis. Among liver tumours there were 19 infantile haemangioendotheliomas and 9 hepatoblastomas. By contrast, at ages 7-24 months hepatoblastomas (n = 28) were much more frequent than haemangioendotheliomas (n = 2). Tumours in the newborn and very young infant differ from those in older children in many respects: (1) the relative frequency; (2) localization; (3) distribution of histological types; (4) degree of differentiation, often associated with a tendency toward maturation but occasionally also with increasing malignancy; and (5) prognosis, which is comparatively favourable. PMID- 2551694 TI - Peripheral neuropathy in the hypereosinophilic syndrome: a case report. AB - We observed a patient with the hypereosinophilic syndrome that showed as a prominent clinical feature peripheral nerve dysfunction. The neuropathy evolved over 4 months and affected sensory and motor functions. Nerve conduction studies and EMG were compatible with axonal neuropathy. Nerve and muscle biopsies revealed severe axonal degeneration with neurogenic atrophy of muscle. Morphometry of peroneal nerve showed marked axonal loss, more prominent in large myelinated fibers. There was no evidence of vasculitis process. Neuropathy is produced by eosinophil-released substances exerting a neurotoxic effect through direct altered vascular endothelial permeability and local mast cell histamine release. PMID- 2551695 TI - Nifedipine and alpha-adrenoceptor function in human finger skin vessels. AB - Experiments were designed to study the effect of the calcium entry blocker nifedipine on contractions evoked by alpha-adrenoceptor activation in human finger skin vessels and to show whether nifedipine could prevent the vasoconstriction induced by local cooling. Vasoconstriction of finger skin vessels was evoked with phenylephrine (a preferential alpha 1-adrenoceptor agonist), B-HT933 (a preferential alpha 2-adrenoceptor agonist) and by local cooling. The alpha-adrenoceptor agonists were administered into the skin by iotophoresis and local cooling was produced with a Peltier element. Nifedipine significantly attenuated the vasoconstrictor responses both to alpha 1- and alpha 2-adrenoceptor stimulation, the effect on alpha 2-mediated responses being more pronounced. It had no effect on the vasoconstriction induced by local cooling of finger skin. PMID- 2551697 TI - Cyclic AMP can enhance mouse B cell activation by regulating progression into the late G1/S phase. AB - This study demonstrates that several substances which raise intracellular adenosine 3',5'-cyclic monophosphate (cAMP) levels in different ways were able to enhance both RNA and DNA synthesis in mouse purified B cells co-stimulated by the protein kinase C activator phorbol 12-myristate 13-acetate and the Ca2+ ionophore ionomycin, while earlier activation events were not modified. These included early changes in cell size and chromatin decondensing demonstrated by light scatter properties at narrow and 90 degrees angles, increase in Ia expression and loss of surface IgD. We concluded that cAMP can up-regulate mouse B cell activation by controlling progression into the late G1 (G1B)/S phase, but not transition from G0 to early G1 (G1A). Furthermore, because cAMP could synergize with ionomycin but not with phorbol 12-myristate 13-acetate to induce RNA and DNA synthesis, we proposed that the cAMP effects in this model may be related to the protein kinase C pathway. PMID- 2551698 TI - Are the catecholestrogens involved in estrogen-induced striatal dopamine receptor supersensitivity? AB - The ability of tamoxifen, an antiestrogen agent, to antagonise the striatal dopamine receptor hyperactivity induced in rats by chronic treatment with 17 beta estradiol and 2-hydroxyestradiol or with two receptor blockers (haloperidol and sulpiride) was compared. It was found that tamoxifen antagonised both the increase in [3H]spiperone binding sites and the stereotyped behaviour induced by apomorphine in animals treated with the two steroids but had no effect in animals receiving the two dopamine blockers. These results run counter to the view that introduction of a catechol group in a steroid molecule is of decisive importance in the induction of striatal dopamine receptor hypersensitivity. PMID- 2551696 TI - Inhibition of erythrocyte aldehyde dehydrogenase activity and elimination kinetics of diethyldithiocarbamic acid methyl ester and its monothio analogue after administration of single and repeated doses of disulfiram to man. AB - The elimination kinetics of the methyl esters of diethyldithiocarbamic acid (Me DDC) and its monothio analogue (Me-DTC) has been compared in ten alcoholic patients after a single oral dose of 400 mg disulfiram (Antabuse). Erythrocyte aldehyde dehydrogenase (ALDH) activity was continuously followed in the subjects until complete inactivation. The relation between individual steady-state concentrations of Me-DTC in plasma, blood acetaldehyde concentration and the dose of disulfiram sufficient to give the disulfiram alcohol reaction was investigated in 12 healthy volunteers treated with increasing doses of disulfiram and then challenged with ethanol. The mean peak plasma concentration of Me-DDC occurred at 1.8 h and its plasma half-life was 6.3 h. The corresponding values for Me-DTC were 3.3 h and 11.2 h, respectively. This suggests oxidative formation of Me-DTC from Me-DDC. In alcoholics with a rapid decrease in erythrocyte ALDH activity (in less than 5 days), the mean peak plasma concentration of Me-DTC was 278 nmol.l-1, whereas the peak concentration was below the detection limit in subjects with a prolonged inactivation time (7-20 days). The healthy volunteers were divided into three groups of four subjects, with clinically valid disulfiram alcohol reactions at 100, 200, and 300 mg doses of disulfiram, respectively. The mean plasma concentrations of Me-DTC at steady-state were proportional to the disulfiram doses given, compared within groups at different doses, and between groups at equal and different optimal doses of the drug. The concentrations of Me-DTC were significantly increased, as compared above, whereas the blood concentrations of acetaldehyde were not significantly increased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551699 TI - Regional variations in binding capacities at mu-, delta- and kappa-opioid sites in membrane suspensions from rabbit brain. AB - The highest maximum binding capacities at the mu-sites of rabbit brain are in the striatum, with intermediate levels in the diencephalon, mesencephalon, cerebellum and cortex and low levels in the pons-medulla and hippocampus. For the delta-site the highest maximum binding capacity is also in the striatum; there are almost equally low levels in the other brain regions. At the kappa-sites the maximum binding capacities are highest in the diencephalon; there are intermediate levels in the cortex and striatum, and low levels in the mesencephalon, cerebellum, hippocampus and pons-medulla. The KD values lack reproducibility; there are no regional variations at the kappa-site as estimated with [3H](-)-bremazocine, but the possibility cannot be excluded that there are regional variations in the KD values for [3H][D-Ala2,MePhe4,Gly-ol5]enkephalin at the mu-site or for [3H][D Ala2,D-Leu5]enkephalin at the delta-site. It may be important to use saturation analysis in future investigations of the distributions of the binding sites. PMID- 2551700 TI - Action of calcitonin gene-related peptide on rat aortic smooth muscle. AB - The effects of rat calcitonin gene-related peptide (CGRP) on superfused rat aortic smooth muscle cells in cell culture were investigated. Exposure of the cells for 10 min to CGRP (10(-7) M), with or without pretreatment with pertussis toxin, stimulated the release of cyclic AMP but not of prostacyclin, as judged by radioimmunoassay of its stable metabolite, 6-keto-PGF1 alpha. Pretreatment of the cells with pertussis toxin did not alter the response to CGRP. The direct action of CGRP on smooth muscle cells and on the CGRP-induced formation of cyclic AMP did not appear to depend on the production of prostacyclin in these vascular smooth muscle cells. PMID- 2551701 TI - Cross-linking of human 125I-beta-endorphin to a mu-delta opioid receptor complex in rat striatum. PMID- 2551702 TI - Comparison of lung virus titers in susceptible and resistant inbred mouse strains against Sendai virus infection. AB - Susceptibility of inbred mouse strains to Sendai virus (Mol strain) infection was studied. Although some mouse strains showed age differences in susceptibility between 3-to 4-week-old and 7-to 8-week-old mice, such age differences in susceptibility were not observed in susceptible DBA/2N and resistant BALB/cA mice. In 7-to 8-week-old mice, remarkable strain differences were observed in mortality and intensity of the lung lesions, but not in lung virus titers and serum antibody, between resistant BALB/cA and susceptible DBA/2N mice. PMID- 2551703 TI - Quantitation of the frequency of immortalization of normal human diploid fibroblasts by SV40 large T-antigen. AB - The mechanism by which SV40 large T-antigen immortalizes human lung fibroblasts is not yet understood, and the frequency with which immortalization occurs is unknown. Here we report detailed studies of the kinetics of immortalization. Approximately 20-50% of individual T-antigen transfected clones of IMR-90 human lung fibroblasts are able to immortalize. The failure of some clones to immortalize is consistent with the hypothesis that in some cells the aneuploidy induced by T-antigen produces extra copies of the chromosome containing a factor that must be inactivated before immortalization occurs. Within immortalization competent clones, the frequency of immortalization is about 3 X 10(-7). This frequency is consistent with a mutational mechanism for the inactivation of the factor that causes crisis in T-antigen expressing cells. PMID- 2551704 TI - Replicative senescence of human skin fibroblasts correlates with a loss of regulation and overexpression of collagenase activity. AB - The atrophy of extracellular matrix is a common event during the aging of connective tissues. In this study, we tested the hypothesis that the altered ability of senescent cells to be modulated by serum growth factors correlated with a loss of regulation of collagenase synthesis. We examined the levels of immunoreactive procollagenase and collagenase inhibitor (the tissue inhibitor of metalloproteinases, TIMP) associated with young and senescent fibroblasts cultured in vitro. Young fibroblasts cultured in low (0.5%) concentrations of fetal bovine serum respond to increased (10%) serum by increasing levels of procollagenase and TIMP beginning 4.0 h after serum stimulation. In contrast, senescent fibroblasts constitutively produce relatively high levels of procollagenase even when cultured in low levels of serum and do not respond to serum stimulation by increasing procollagenase synthesis. In addition, senescent fibroblasts constitutively express a relatively small amount of TIMP which is not induced upon serum stimulation. This altered expression of collagenase and TIMP appears unique to the senescent phenotype and not merely a result of growth inhibition, since young cells growth arrested by density-dependent growth inhibition displayed a temporal pattern of procollagenase and TIMP expression upon serum stimulation similar to that of subconfluent young cultures. An assay of net collagenase activity revealed a greater than 20-fold elevation of activity in trypsin-activated extracts from senescent versus young fibroblasts when cultured in a low concentration of fetal bovine serum. These results demonstrate for the first time a direct correlation between alterations in the molecular pathways regulating connective tissue homeostasis and those of replicative senescence. The increased collagenolytic activity of senescent compared to young fibroblasts cultured in the presence of a low serum concentration suggests that aging fibroblasts may become increasingly fibroclastic causing many of the age associated alterations in dermal collagen observed during aging in vivo. PMID- 2551705 TI - Forskolin inhibits the Gs-stimulated adenylate cyclase in rat ascites hepatoma AH66F cells. AB - Forskolin increased intracellular cyclic AMP and augmented cyclic AMP formation by prostaglandin E1 (PGE1) in normal rat hepatocytes and ascites hepatoma AH66 cells. However, in AH66F cells which were derived from the AH66 cell line, the diterpene only slightly increased the cyclic AMP level, and dose-dependently inhibited the accumulation caused by PGE1. Forskolin dose-dependently activated adenylate cyclase in these membranes, and the magnitude of activation by forskolin was largest in the following order: hepatocytes, AH66 cells, and AH66F cells. This difference may be based on the number of forskolin-binding sites. The binding affinity of forskolin for each cell membrane was similar. The number and affinity of forskolin-binding sites in these cells were not influenced by 5' guanylylimidodiphosphate [Gpp(NH)p]. In hepatocytes and AH66 cells, forskolin and other adenylate cyclase activators such as PGE1, GTP, Gpp(NH)p, F-, and Mn2+ synergistically increased the enzyme activity. In AH66F cells, the forskolin stimulated activity was hardly influenced by the GTP analog, and forskolin diminished the activities induced by the GTP analog in a manner similar to that of diterpene alone. Forskolin (10 microM) also significantly inhibited the activities induced by PGE1, GTP, and F-. The effect of forskolin with Mn2+ was additive in AH66F cells. The data suggest that forskolin promotes the interaction between the stimulatory guanine nucleotide-binding protein and the catalytic unit in the membrane of normal hepatocytes and AH66 cells, but it interferes with the coupling in AH66F cells. PMID- 2551706 TI - Preclustered receptor arrangement is a prerequisite for galactose-specific clearance of large particulate ligands in rat liver. AB - We had hypothesized that preclustered arrangement of galactose-specific receptor activity on rat liver macrophages enables these cells to internalize multivalent, particulate ligands in contrast to the clearance of molecules mediated by statistically distributed receptors on hepatocytes. We now took advantage of the nonclustered receptor distribution in newborn rat liver macrophages to study the in vivo clearance of particulate ligands. Gold particles 5, 17, and 50 nm in diameter (Au5, Au17, Au50), coated with lactosylated bovine serum albumin (LacBSA), were injected into the vena cava and livers were perfusion fixed after allowing for binding and uptake for 3 min. In sinusoidal cells from rats 15 days old LacBSA-Au5 and LacBSA-Au17 were taken up by endothelial cells and all sizes by liver macrophages. In newborn rat liver no LacBSA-Au50 or LacBSA-Au17 was retained in liver macrophages. Uptake of LacBSA-Au5 by sinusoidal cells was significant. LacBSA-Au17 was taken up in significant amounts by endothelial cells of newborn rats which correlates to the findings that galactose-specific binding sites on endothelial cells were found to localize as clusters over coated pits irrespective of age. These results demonstrate the crucial role of clustered receptors in binding and uptake of larger particulate ligands via this lectin like binding activity. PMID- 2551707 TI - Interactive effects of Na and K in killing by human natural killer cells. AB - Contact-mediated lysis by human natural killer cells is inhibited by a number of drugs that block the predominant K channel. In this study we have further examined the role of the K channel and the interactions between passive K and Na transport in killing. Low external Na-inhibited killing and inhibition were not due to reduce inward current through the Na channels in the target cell. A role for the Na/H antiport is suggested since amiloride inhibited killing in a dose dependent manner that was competitive with external Na. Depolarizing the killer cell with elevated external K did not inhibit killing. On the contrary, high K0 reduced the inhibition caused by low Na0 and by the K-channel blockers quinidine, verapamil, and retinoic acid. Hyperpolarizing the killer cell with low K0 or valinomycin inhibited killing. Valinomycin, which should prevent the depolarization caused by K-channel block, did not reverse the effect of the blockers quinidine, verapamil, and 4-aminopyridine. Hence, the primary role of the K channels during killing is not maintain the negative membrane potential. On the contrary, depolarization may promote killing under conditions where killing is submaximal. PMID- 2551708 TI - Isolation of rod outer segments on Percoll gradients: effect of specific protease inhibition. AB - Rat rod outer segments (ROSs) were isolated by vortexing retinas and separating the detached components on performed Percoll gradients. A lighter band of 20 x 10(6) unsealed ROSs per ten retinas, and a heavier band of 60 x 10(6) sealed ROSs per ten retinas were obtained from each 12 ml gradient. The yield of sealed ROSs (but not unsealed ROSs) was increased up to twofold in the presence of the specific cysteine protease inhibitor, Ep-475. Aprotinin, pepstatin, PMSF, TPCK and EGTA plus EDTA had no effect. These results indicate that during isolation, ROSs are vulnerable to damage by cysteine protease activity either from damaged retinal cells or from within. PMID- 2551709 TI - Unitary characteristics of presumptive cholinergic tegmental neurons during the sleep-waking cycle in freely moving cats. AB - A total of 260 neurons were recorded in the rostral pontine tegmentum of freely moving cats during the sleep-waking cycle. Of these, 207 neurons (80%) were located in the dorsal pontine tegmentum containing monoaminergic and choline acetyltransferase (ChAT)-immunoreactive, or cholinergic neurons. In addition to presumably monoaminergic PS-off cells (n = 51) showing a cessation of discharge during paradoxical sleep (PS) and presumably cholinergic PGO-on cells (n = 40) exhibiting a burst of discharge just prior to and during ponto-geniculo-occipital (PGO) waves, we observed tonic (n = 108) and phasic (n = 61) neurons exhibiting, respectively, tonic and phasic patterns of discharge during wakefulness and/or paradoxical sleep. Of 87 tonic cells histologically localized in the dorsal pontine tegmentum rich in cholinergic neurons, 46 cells (53%) were identified as giving rise to ascending projections either to the intralaminar thalamic complex (n = 26) or to the ventrolateral posterior hypothalamus (n = 13) or to both (n = 9). Two types of tonic neurons were distinguished: 1) tonic type I neurons (n = 28), showing a tonic pattern and high rates of discharge during both waking and paradoxical sleep as compared with slow wave sleep; and 2) tonic type II neurons (n = 20), exhibiting a tonic pattern of discharge highly specific to the periods of paradoxical sleep. Tonic type I neurons were further divided into two subclasses on the basis of discharge rates during waking: a) rapid (Type I-R; n = 17); and b) slow (Type I-S; n = 11) units with a discharge frequency of more than 12 spikes/s or less than 5 spikes/s, respectively. Like monoaminergic PS-off and cholinergic PGO-on cells, both tonic type II and type I-S cells were characterized by a long spike duration (median: 3.3 and 3.5 ms), as well as by a slow conduction velocity (median = 1.8 and 1.7 m/s). In the light of these data, we discuss the possible cholinergic nature and functional significance of these ascending tonic neurons in the generation of neocortical electroencephalographic desynchronization occurring during waking and paradoxical sleep. PMID- 2551710 TI - Firing properties of respiratory rhythm generating neurons in the absence of synaptic transmission in rat medulla in vitro. AB - It has previously been demonstrated that Pre-I neurons, localized in the rostral ventrolateral medulla, are important in the generation of the primary respiratory rhythm in brainstem-spinal cord preparations from newborn rats. To investigate whether or not Pre-I neurons have endogenous pacemaker properties, we examined Pre-I neuron activity before and after chemical synaptic transmission was blocked by incubation in a low Ca2+ (0.2 mM), high Mg2+ (5 mM) solution (referred to here as low Ca). After incubation for about 30 min in low Ca, 28 (52%, type-1) out of 54 neurons tested in 27 preparations retained apparent rhythmic (phasic) activity after complete disappearance of C4 inspiratory activity. Sixteen neurons (30%, type-2) fired tonically and 10 (18%, type-3) were silent. We examined the effects of synaptic blockade on 14 inspiratory neurons in the RVL. The firing of all 14 neurons in 9 preparations disappeared concomitantly with the disappearance of C4 activity in low Ca. When the pH of the low Ca solution was lowered with a decrease in NaHCO3 concentration from 7.4 to 7.1, the firing rate of the Pre-I neurons (type-1) increased from 12 to 18/min. In conclusion, the generator of respiratory rhythm in the newborn rat is probably a neuronal network with chemical synapses that functions mainly through the endogenous Pre-I pacemaker cells. Intrinsic chemoreception in the rhythm generator is probably important in frequency control of respiratory rhythm. PMID- 2551711 TI - Interactions between natural and electrically evoked saccades. I. Differences between sites carrying retinal error and motor error signals in monkey superior colliculus. AB - Fixed-vector saccades evoked by electrical stimulation may result from the elicitation of a retinal error signal directing the eyes toward a goal, or from the elicitation of a motor error signal determining the vector itself. Theoretically, the two mechanisms can be differentiated by delivering the stimulation while the eyes are already in motion (colliding saccade paradigm), thereby changing the eye position from which the evoked saccade starts. Only in the first case is the trajectory of the evoked saccade expected to be modified to compensate for part of the ongoing eye movement. An attempt was made to distinguish retinal vs. motor error mechanisms by applying the colliding saccade paradigm of stimulation to 29 sites throughout the superior colliculus (SC) of two trained monkeys. Compensatory evoked saccades, as predicted by the retinal error hypothesis, were obtained consistently in the superficial layers and at deeper sites where visual unit responses could be recorded. Conversely, in deep layers where only presaccadic activity was found, evoked saccades either were not affected by collision or summed their vectors with that of the ongoing movement. These last observations are both consistent with the hypothesis that the signal produced from deep sites was an initial motor error. A second observation was incidentally made: when stimulation was applied to the most superficial SC region, it definitively erased the goal of the ongoing saccade, and the latter did not resume its interrupted course. The colliding saccade paradigm may be useful in clarifying the role of structures involved in oculomotor function. PMID- 2551712 TI - Interactions between natural and electrically evoked saccades. II. At what time is eye position sampled as a reference for the localization of a target? AB - Electrical microstimulation was applied at brain sites (thalamic internal medullary lamina complex and superficial layers of superior colliculus) of alert, trained monkeys to evoke fixed-vector saccades. When the stimulation was timed to occur during or after an eye movement, the evoked saccade had a modified trajectory, compensating for, at least, the last portion of the ongoing eye movement. The hypothesis proposed to explain this compensatory effect (Schlag-Rey et al. 1989) is that the electrical stimulation produces a saccade by generating a signal, equivalent to a retinal error, specifying the saccade goal at a fixed location with respect to some eye position (called reference eye position). If the eyes are moving at the time of stimulation, the reference eye position lies somewhere along the trajectory of the ongoing movement. In the present study, we tried to determine this reference eye position, and deduce from it the instant at which the goal was specified. A significant timing difference was observed between thalamic and collicular stimulations. The goal appeared to be referred to an eye position existing at stimulation onset in superior colliculus (SC), and 35 65 ms before stimulation onset in central thalamus. In the latter case, the results suggest that the evoked saccade was aimed at the spatial location that the brain computed by summing a retinal error signal (evoked by stimulation) with the eye position at the time such a signal would have been elicited by a real target. In contrast, the collicular results suggest that the evoked saccade was directed to the retinal location specified by the retinal error signal. The findings imply that if the eyes are not steady while the target position is calculated, signals conveyed in the superficial layers of SC (in contrast to the thalamus) cannot direct the eyes correctly to a visual target. PMID- 2551713 TI - Somatosensory areas in the telencephalon of the pigeon. I. Response characteristics. AB - Two somatosensory regions in the pigeon's telencephalon were investigated electrophysiologically with recordings of field potentials as well as single- and multi-unit responses which were evoked by electrical stimulation of all four extremities or by feather movements produced with airpuffs or by hand. The outline of both areas, was studied in detail with the use of grid-like recordings of single or multi-units. One somatosensory area is located rostrally in the hyperstriatum accessorium (HA), rostral to the visual "Wulst". A caudal area comprises the medial aspects of two different cell layers: the neostriatum intermedium (NI) and adjacent neostriatum caudale (NC) as well as the overlying hyperstriatum ventrale (HV). The two areas differ considerably in their response characteristics. Field potentials of the NI/NC-HV area were more complex than those of the HA area and their shapes and latencies varied mainly in dependence of the recording site (NI, NC, HV). Multi-unit responses showed strong excitation and short latencies in NI/NC and weak excitation and longer latencies in HV. Both responses and latencies were uniform in the HA area and latencies generally longer than in NI/NC but shorter than in HV. The HA area processes somatosensory information more specifically. Its neurons have relatively small receptive fields which seem to be arranged in a somatotopic order in such a way that rostral parts of the body are represented superficially and caudal parts in deeper layers. In contrast, the NI/NC-HV area was found to be largely multimodal, receiving also auditory and visual information. Neurons in this region have large somatic receptive fields, often including one and sometimes even both sides of the body surface. A somatotopic arrangement could not be recognized. The whole body surface was representated in both areas, but there was a dominance of wing and back receptive fields in the NI/NC-HV area and leg and neck receptive fields in the HA area. PMID- 2551714 TI - Propagation velocity of epileptiform activity in the hippocampus. AB - The propagation of epileptiform burst activity was investigated in the CA1 area of the in-vitro hippocampal slice preparation of the guinea pig. This activity was provoked by 0.1 mM 4-aminopyridine in the bathing medium and was recorded in the pyramidal layer with an array of eight electrodes. The delay between the first population spike of a burst recorded with different electrodes was calculated using the cross-correlation function. The propagation velocity was estimated from the delays and the electrode intervals. It was found that the velocity of spontaneous and evoked epileptiform bursts varies between 0.15 and 5 m/s and is not confined to the range of conduction velocities of the fibre systems in CA1 (0.3-0.55 and 1.0-1.8 m/s). Different velocities can be present in different parts of the CA1 area and the initiation of spontaneous bursts is not confined to the CA2-3 areas, but can also occur in CA1. Burst activity also propagated in a low calcium-high magnesium medium. Different mechanisms of propagation are discussed and it is argued that the propagation velocity due to ephaptic interaction may vary largely. It is concluded that epileptiform activity can be propagated not only by synaptic connections at or near the pyramidal layer, but also by way of electrical field effects of population spikes. PMID- 2551715 TI - Anaesthetic depression of excitatory synaptic transmission in neocortex. AB - A decrease in synaptic excitation as well as an enhancement of neuronal inhibition in the central nervous system have been suggested as possible mechanisms of anaesthesia which we have investigated with intraneuronal recording techniques in neocortex. The effects of a volatile agent--isoflurane and a steroid preparation--Althesin, on excitatory and inhibitory postsynaptic potentials (EPSPs and IPSPs) evoked by epicortical electrical stimulation were investigated in in vitro slice preparations of anterior cingulate and sensorimotor cortices of guinea pig. Applications of isoflurane (0.5-2.5 minimum alveolar concentration or MAC) and Althesin (10-200 microM) induced a dose dependent, reversible depression of EPSPs with EC50's of 1 MAC and approximately 50 microM respectively. In order to eliminate the possibilities of a shunting effect on EPSPs by the conductances involved in the concomitant IPSPs, a GABAA antagonist (bicuculline) was applied together with the anaesthetics. With this IPSP blockade, both anaesthetics depressed the EPSPs and were effective in reducing the epileptiform activities evoked by bicuculline during the subpial stimulation. The IPSPs also were evoked during the blockade of K-conductances by internal Cs-applications in order that the effects of anaesthetics could be studied exclusively on the Cl-mediated GABAergic IPSPs. Both isoflurane (0.5-2.5 MAC) and Althesin (10-100 microM) depressed the IPSPs in a dose dependent manner. These investigations demonstrate that applications of isoflurane and Althesin depressed the excitabilities of neocortical neurons by interfering with synaptic excitation, possibly at pre- and postsynaptic sites, rather than by potentiating neuronal inhibition. PMID- 2551716 TI - Longer term progesterone treatment induces changes of GABAA receptor levels in forebrain sites in the female hamster: quantitative autoradiography study. AB - Quantitative receptor autoradiography was applied to evaluate the effects of one and three injections of 1 mg progesterone (P) on 3H muscimol binding levels in the different forebrain areas of the female hamster. The overall effect of P resulted in substantial increases in 3H muscimol binding in brain areas containing gonadal steroid receptors: medial preoptic area and ventromedial hypothalamic nucleus as well as in bed nucleus stria terminalis and subiculum. Similarly, the caudate putamen, a region where gonadal steroid receptors are not abundant, also showed substantial increases of 3H muscimol binding receptor levels. Moreover, female hamsters treated with P for 3 days presented altered 3H muscimol binding levels in the amygdala and thalamic nucleus that were, in some cases, not produced by one dose of P. P treatment also decreased GABAA binding in two areas of the thalamus. These results are consistent with the proposal that P may alter GABAergic inhibitory activity via changes in the levels of GABAA receptors in certain forebrain areas in the female hamster, changes which may be linked to the mediation of anxiolytic effects and to the inhibition of aggressive behavior. These data also suggest that P treatment increases the binding of high affinity GABA receptors in some forebrain sites and may be responsible for maintenance of the anxiolytic effects. PMID- 2551717 TI - Calcineurin in the postnatal striatum of the rat: an immunohistochemical study. AB - This report concerns the postnatal expression of calcineurin (CaN) in rat striatum. For this purpose an immunohistochemical procedure was used. In neonatal striatum CaN immunoreactivity was found in discrete patches composed of many immature cells. With development there was an increase in staining intensity and in the proportion of positively reacting cells. CaN expression and localization in the striatum of 22-25 day old rats were similar to those of adult animals. PMID- 2551718 TI - Extracellular potassium activity and axonal conduction in spinal cord of the myelin-deficient mutant rat. AB - We recorded somatosensory evoked potentials SEPs), extracellular K+ ionic activity ([K+]e), and K+ clearance rates in the spinal cords of 14 myelin deficient mutant rats and 16 normal male littermates at 16-41 days after birth. Tested under pentobarbital anesthesia (25 mg/kg ip) and hypothermic conditions (32-34 degrees C), myelin-deficient rats had longer cortical SEP latencies (67 +/ 20 ms) compared to those in normal siblings (48 +/- 15 ms; P less than 0.05). Mean baseline [K+]e levels were 2.6 +/- 0.5 mM in myelin-deficient rats and 2.6 +/- 0.8 mM in normal siblings. Clearance times of KCl solutions injected into the spinal cord were biphasic and exponential. The mean initial and secondary exponential half-times were 1.0 +/- 0.5 and 2.7 +/- 1.7 min for myelin-deficient rats and 0.8 +/- 0.4 and 3.8 +/- 3.2 min for normal siblings. Repetitive sciatic nerve stimulation (2-20 Hz, 2- to 6-s trains) produced 1-3 mM transient [K+]e rises in thoracic and lumbar cords of myelin-deficient rats. The [K+]e rises were largest in the dorsal spinal cord at 200-500 microns depth. The normal siblings had smaller or no stimulus-induced [K+]e rises. In myelin-deficient rats, injection of 1 mM 4-aminopyridine (4-AP) solution into the thoracic spinal cord completely suppressed the stimulus-induced [K+]e and markedly increased spinal and cortical SEP amplitudes for several hours. In the normal siblings, the 4-AP injections transiently blocked spinal conduction for 20-30 min but thereafter enhanced cortical SEP amplitudes for 2-3 h. We conclude that sciatic nerve stimulation produces spinal cord [K+]e rises in myelin-deficient rat larger than those in the normal siblings, that the [K+]e transients represent increased K+ release rather than impaired K+ clearance, and that the K+ ions come from 4-AP blockable sources. PMID- 2551719 TI - Studies on the potential neurotoxic and convulsant effects of increased blood levels of quinolinic acid in rats with altered blood-brain barrier permeability. AB - Intravenous injection of 450 mg/kg quinolinic acid (Quin), an endogenous kynurenine metabolite with excitotoxic properties, induced only minor electroencephalographic (EEG) modifications and no neurotoxicity in rats with a mature blood-brain barrier (BBB). BBB permeability was altered in rats by focal unilateral irradiation of the cortex (7 mm in diameter and 5 mm in depth) with protons (60 Gy, 9 Gy/min). Three days after irradiation, Evans blue dye staining showed BBB breakdown in the dorsal hippocampus of the irradiated hemisphere. No neurotoxic or convulsant effects were observed as a consequence of the radiation itself. When BBB-lesioned rats were challenged with 225 mg/kg Quin iv, epileptiform activity was observed on EEG analysis. Tonic-clonic seizures were induced by 225-450 mg/kg Quin. Light microscopic analysis showed a dose-related excitotoxic type of lesion restricted to the hippocampus ipsilateral to the irradiated side. Neuro-degeneration was prevented by local injection of 120 nmol D(-)2-amino-7-phosphonoheptanoic acid, a selective N-methyl-D-aspartate receptor antagonist. No lesions or EEG or behavioral modifications occurred after 450 mg/kg nicotinic acid, an inactive analog of Quin. The potential neurotoxic and convulsant effects of increased blood levels of Quin under conditions of altered BBB permeability are discussed. PMID- 2551720 TI - Thermal properties of red blood cells infected by malaria parasites. AB - Three membrane thermotropic transitions at 8, 20, and 40 degrees C have been detected in human red blood cells (RBC) by using spin-labeled stearic acids. Red blood cells infected in vitro by Plasmodium falciparum showed the disappearance of the 8 degrees C transition and a lowering of the 40 degrees C transition to 32 degrees C. The disappearance of the 8 degrees C transition was observed in synchronized cultures of P. falciparum trophozoites as well as in mouse RBC infected in vivo by an asynchronous population of P. berghei. Furthermore, erythrocytes infected by P. falciparum showed an increase in the phosphorylation of protein 4.1. This protein was shown previously to be involved in the 8 degrees C transition, (T. Forte, T. L. Leto, M. Minetti, and V. T. Marchesi, Biochemistry 24, 7876-7880 (1985). Our results suggest that the malaria parasite invasion produces a disorganization of the protein 4.1-membrane interaction. PMID- 2551721 TI - Molecular pathogenesis of type 2 poliovirus in mice. AB - Lansing strain of poliovirus type 2 (PV-2) produces a fatal paralytic disease in mice after intracerebral inoculation. To identify virus-containing nerve cells in poliovirus-infected mice, we developed a technique of in situ hybridization for viral RNA with a poliovirus-specific riboprobe. Large numbers of genomes were found in motoneurons and their processes, as well as in ghosts of neurons with or without inflammatory cells. This indicates that the death of motoneurons is due to a direct effect of viral replication. The presence of viral RNA in neuronal processes suggests an axonal dissemination of poliovirus within the central nervous system (CNS). Detection of viral genomes in small cells located in anterior and in posterior horns shows that motoneurons are not the only cells susceptible to poliovirus in the CNS. To see if PV-2/Lansing still exits in nature, PV-2 isolated from the CNS and from the gut of two human cases of paralytic poliomyelitis were characterized by determining epitope maps and phenotypic markers and by sequencing regions of the genome coding for antigenic sites and the 5' non-coding region. Comparison of virus isolated from the intestine and from the CNS of the same subject revealed a high degree of homology, suggesting that virus isolated from the gut is representative of virus in the neuronal lesions. The isolate from a case had more homology to PV 2/Lansing, whereas the other isolate showed more homology to PV-2/Sabin. These results show that, despite intensive vaccination with oral polio vaccine for 25 years, wild PV-2/Lansing-like viruses, pathogenic for mice, still circulate in nature ad induce human paralytic cases. PMID- 2551722 TI - Immunological status to Epstein-Barr virus and cytomegalovirus in patients with genital condylomata. AB - Serological patterns against Epstein-Barr virus (EBV) and Cytomegalovirus (CMV) were determined in patients with genital condylomata (GC). The Ig G antibody values to EB-induced virus capsidic antigens (VCA), early antigens (EA) and Ig M to VCA were significantly higher in the study group than in the controls. Moreover, the concomitant presence of EBV-Ig G anti-VCA greater than or equal to 1/320, EBV-Ig G anti-EA greater than or equal to 1/20 and EBV-Ig M anti VCA greater than or equal to 1/20 was observed in 13 serum samples of genital condylomata patients, while, in only 2 serum samples of the healthy controls, the same serological pattern was present. The distribution of antibody values to CMV induced LA, EA and IEA showed a significantly increased prevalence in the study group in comparison with the controls: the concomitant presence of antibody with a titre greater than or equal to 1/320 for CMV-LA, greater than or equal to 1/20 for CMV-EA and greater than or equal to 1/20 for CMV-IEA was observed in 15 serum samples of GC patients and in only 3 serum samples of the control group. Our results suggest that the active or recent EBV and CMV infections we observed in genital condylomata patients may be a consequence of impaired immunity in these patients, but it does not exclude a possible role of EBV and CMV in perpetuating human papilloma virus-induced cell proliferation. PMID- 2551723 TI - Coxsackievirus B3 myocarditis in C3H/HeJ mice: description of an inbred model and the effect of exercise on virulence. AB - The effect of forced exercise on the development of coxsackievirus B3 myocarditis in inbred C3H/HeJ mice was studied. Four groups of mice (30 per group) were formed: infected-exercised (Group I); infected-unexercised (Group II); uninfected exercised (Group III); and uninfected-unexercised (Group IV). Infected mice were inoculated intraperitoneally with 1.0 x 10(2.1) TCID50 coxsackievirus B3. Exercised animals were swum daily for 60 minutes on days 1-9. Myocardial viral titers were acutely elevated on day 3 of infection and were augmented significantly by exercise on days 6 and 9. Exercise increased the overall mortality from 0-10% to 20-40%; significantly increased heart: body weight ratios on days 6, 9 and 13; and increased the extent of myocardial fiber necrosis. We have reproduced the acceleration of CB3 myocarditis by exercise in the inbred C3H model. PMID- 2551724 TI - [Participation of opioids and opioid receptors in antinociception at various levels of the nervous system]. AB - At the cerebral level, studies employing several experimental approaches point to an essential role of beta-endorphin in analgesia, induced by electrical stimulation of the periaqueductal grey of the midbrain. Tolerance and cross tolerance studies suggest that mu-opioid receptors mediate this effect. The significance of sigma- and, in particular, kappa-opioid receptors in cerebral pain modulation is less well documented. At the spinal level, nociception is relayed in the dorsal horn, where opioid peptides as well as all types of opioid receptors are abundant. mu-opioid receptor-mediated anti-nociceptive processes appear to be most important in this region, but sigma-opioid receptors may also be involved. In addition, a role of kappa-opioid receptors can be demonstrated under certain conditions. Recent experiments indicate that opioids may also modulate nociception in the periphery, in particular in inflamed tissue. PMID- 2551725 TI - Energy transduction at the catalytic site of enzymes: hydrolysis of phosphoester bonds and synthesis of pyrophosphate by alkaline phosphatase. AB - Alkaline phosphatase from mouse intestinal epithelial cells catalyzes the synthesis of pyrophosphate from Pi during hydrolysis of either glucose 6 phosphate, ATP, ADP, inorganic pyrophosphate or p-nitrophenylphosphate. The rate of pyrophosphate synthesis is increased by MgCl2 and by decreasing the pH of the medium from 8.5 to 6.0. The data presented indicate that at the catalytic site of alkaline phosphatase the energies of hydrolysis of the phosphoserine residue and of pyrophosphate are different from those measured in aqueous solutions. PMID- 2551726 TI - Presence of fructokinase in pancreatic islets. AB - Homogenates of rat pancreatic islets that had been heated for 5 min at 70 degrees C to inactive hexokinases, catalyzed the ATP-dependent phosphorylation of D fructose. This reaction was dependent on the presence of K+ and was inhibited by D-tagatose although not by D-glucose or D-glucose 6-phosphate. The phosphorylation product was identified as fructose 1-phosphate through its conversion to a bisphosphate ester by Clostridium difficile fructose 1-phosphate kinase. These findings allowed the conclusion that fructokinase (ketohexokinase) was responsible for this process. Similar results were observed with tumoral insulin-producing cells (RINm5F line). Fructokinase may account for a large share of fructose phosphorylation in intact islets, particularly in the presence of D glucose. PMID- 2551727 TI - Inhibition by islet-activating protein, pertussis toxin, of retinoic acid-induced differentiation of human leukemic (HL-60) cells. AB - Human promyelocytic leukemic (HL-60) cells were induced to differentiate into neutrophil- or macrophage-like cells by incubation of the cells with retinoic acid, dibutyryl cyclic AMP (Bt2cAMP) or phorbol 12-myristate 13-acetate (PMA). Differentiation was determined by an increase in the percentage of morphologically mature cells. The retinoic acid-induced differentiation of HL-60 cells was, but the Bt2cAMP- or PMA-induced one was not, inhibited by prior exposure of the cells to islet-activating protein (IAP), pertussis toxin. The IAP induced inhibition was correlated with the toxin-catalyzed ADP-ribosylation of a membrane GTP-binding protein with a molecular mass of 40 kDa. Thus, the IAP substrate GTP-binding protein appears to be involved in the retinoic acid-induced differentiation of HL-60 cells. PMID- 2551728 TI - Effect of the different dimeric forms of the platelet-derived growth factor on cellular responses in mouse Swiss 3T3 fibroblasts. AB - PDGF consists of two polypeptide chains, A and B, and all three possible dimers have been isolated from different sources. Human PDGF, essentially AB, porcine PDGF (BB) and recombinant PDGF-AA were tested on Swiss 3T3 fibroblasts for their ability to stimulate mitogenesis, phosphoinositide turnover and tyrosine phosphorylation of the PDGF receptor. When used in saturating amounts, the three isoforms were equally active in inducing mitogenesis. However, PDGF-AA was less active than AB and BB to induce the phosphorylation of the receptor and the turnover of phosphoinositides (30% and 50%, respectively). These findings suggest that, in Swiss 3T3 fibroblasts, PDGF receptors of the alpha-type are present in a slightly lower amount than beta-type. In addition, the two types of receptor appear to have similar physiological functions. PMID- 2551729 TI - The N-terminal domain I of human lactotransferrin binds specifically to phytohemagglutinin-stimulated peripheral blood human lymphocyte receptors. AB - Human lactotransferrin receptors have been recently characterized on mitogen stimulated human lymphocytes [(1989) Eur. J. Biochem. 179, 481-487]. In order to define the lactotransferrin recognition site by these receptors, the binding to lymphocytes of several tryptic fragments, isolated from human lactotransferrin by mild tryptic hydrolysis [(1984) Biochim. Biophys. Acta 787, 90-96], has been investigated. The 30 kDa N-tryptic fragment (residues 4-281) and the re associated N,C-tryptic complex bind to lactotansferrin lymphocyte receptor with a dissociation constant of 44 nM and 39 nM, respectively, similar to the value obtained for the native lactotransferrin (Kd = 46 nM). However, neither the N terminal domain II (residues 91-257) nor the 50 kDa C-tryptic fragment (residues 282-703) are recognized. These results suggest that the binding site of human lactotransferrin by the lymphocyte receptor is located in the N-terminal lobe and more precisely in the N-terminal domain I (residues 4-90 and/or 258-281). PMID- 2551730 TI - Molecular cloning and sequence analysis of cDNA encoding rat adrenal cytochrome P 450(11)beta. AB - A cDNA clone encoding cytochrome P-450(11)beta of rat adrenal has been cloned and sequenced using a bovine P-450(11)beta cDNA insert (pcP-450(11 beta)-2; (1987) J. Biochem. 102, 559-568) as a probe. The nucleotide sequence contains an open reading frame sufficient to encode the entire amino acid sequence of a P 450(11)beta precursor protein consisting of 499 amino acids including an extension peptide of 24 amino acids at the NH2-terminus. The cDNA contains 1247 nucleotides at the 3'-noncoding region including 51 nucleotides of poly A, but lacks the 5'-noncoding region. The deduced amino acid sequence shows 61% similarity to that of bovine P-450(11)beta. Putative binding sites for heme and steroid are highly conserved among steroidogenic P-450s of known structure. PMID- 2551731 TI - Electron-transfer restoration by vitamin K3 in a complex III-deficient mutant of S. cerevisiae and sequence of the corresponding cytochrome b mutation. AB - The yeast box-mutant W7 exhibits deficiencies in cytochrome b and in nuclear coded complex III subunits, a phenotype observed previously in a patient with mitochondrial myopathy. DNA sequence analysis of mutant W7 revealed a single base transition in the cytochrome b gene; the mutated residue Gly 131 is perfectly conserved in all known cytochromes b and belongs to the Qo domain. Mutant W7 provides a model system for evaluating the action of therapeutic agents, such as vitamin K3 which restored NADH-oxidase activity in the mutant as well as in the antimycin-inhibited wild type. However, with the mutant, a greater quantity of menadione was necessary due to a decrease in other complex activities, and a much lower electron-flow fraction passed through cytochrome oxidase. PMID- 2551732 TI - Isolation from human cerebrospinal fluid of a new insulin-like growth factor binding protein with a selective affinity for IGF-II. AB - In biological fluids IGF-I and IGF-II are bound to specific, high-affinity binding protein (BPs). Two human BPs have been isolated, one from serum, which is GH-dependent, the other from amniotic fluid (AF BP), and their cDNAs have recently been cloned. We report here the isolation of another, new species from cerebrospinal fluid (CSF) where this BP predominates. The protein was purified to homogeneity by a four-step procedure: gel filtration, chromatofocusing, hydrophobic-interaction chromatography and reverse-phase chromatography. Thereafter, SDS-polyacrylamide gel electrophoresis gave an Mr of 34,000 (non reduced), chromatofocusing gave an isoelectric point of 5.0m and its affinity for IGF-II (3 x 10(10) M-1) was 10 times that for IGF-I. The N-terminal amino acid sequence of the first 15 residues determined in a BP preparation from the CSF of children was Leu-Ala-Pro-Gly-(/)-Gly-Gln-Gly-Val-Gln-Ala-Gly-Ala-Pro-Gly. A similar sequence was found for adult CSF, apart from residues 12 and 13 (-Leu-Leu ). These are highly analogous with the sequences starting from residue 69 of the GH-dependent BP, and from residue 61 of the AF BP. The new BP isolated is therefore related to, but distinct from, the other human BPs. PMID- 2551733 TI - Analysis of free sulfhydryl groups and disulfide bonds in Na+,K+-ATPase. AB - The content of free SH groups and disulfide bonds in the purified pig kidney Na+,K+-ATPase was determined by ammetric titration with silver nitrate. In the native enzyme, most of the free SH groups are masked due to their location in the polypeptide chain regions poorly accessible to SH reagents. Denaturation with 5% SDS and 8 M urea makes these regions accessible thus revealing 22 free SH groups/mol of the protein. After complete blocking of free SH groups with silver ions, 8 SH groups/mol of the protein are being released upon sulfitolysis which indicates the presence of four disulfide bonds in the enzyme. At least one disulfide bridge is located in the alpha-subunit whereas the beta-subunit contains three disulfide bonds. PMID- 2551735 TI - Human gelatinase/type IV procollagenase is a regular plasma component. AB - Gelatin zymograms revealed in human plasma a constant 66 kDa proteolytically active polypeptide. In most plasma samples other major proteolytic activities were seen at Mr 92,000, 130,000 and 225,000. All four proteases were Ca2+ dependent metalloproteases and bound quantitatively to gelatin-Sepharose. Immunoblotting results indicated that the 66 kDa protease was the human fibroblast gelatinase/type IV procollagenase and that the other three proteases were macrophage/granulocyte-derived gelatinase components. The 66 kDa protease did not bind to conA- nor lentil lectin-Sepharose allowing its separation from the 92, 130 and 225 kDa proteases. During the isolation procedure the plasma gelatinase/type IV procollagenase tended to form a proteolytically active spontaneous disulfide-bonded dimer and a 62 kDa component that could also be obtained by digestion with trypsin. The same polypeptide changes occurred also in stored preparations of the corresponding protease isolated from fibroblast culture medium while the freshly purified protein contained only the 66 kDa proform. PMID- 2551734 TI - Isolation of dermenkephalin from amphibian skin, a high-affinity delta-selective opioid heptapeptide containing a D-amino acid residue. AB - The predicted amino acid sequence of the biosynthetic precursor of dermorphin, a highly potent and nearly specific mu-opioid peptide from amphibian skin, contains four repeats of the dermorphin progenitor sequence and one single copy of a different heptapeptide sequence. We have developed a specific enzyme immunoassay and used synthetic peptides to detect and purify the new predicted heptapeptide (2.4 micrograms/g dry skin) from the skin of the Phyllomedusa sauvagei frog from which dermorphin was originally isolated. The identity of the novel pro dermorphin related peptide, Tyr-D-Met-Phe-His-Leu-Met-Asp-NH2, was established by co-chromatography with synthetic peptides on reverse-phase HPLC, immunological analysis, gas-phase sequencing, mass spectrometry and by pharmacological assays. Opioid-binding assays in vitro demonstrated that both the natural and synthetic heptapeptides displayed exceptionally high selectivity and affinity towards the delta-opioid receptors. Because of its origin and its delta-opioid (enkephalin) activity and specificity, this novel D-amino acid containing peptide is named dermenkephalin. PMID- 2551736 TI - Effect of subtransition in 1,2-dipalmitoylphosphatidylcholine model membrane on the spin probe motion. AB - The effect of subgel----gel phase transition (subtransition) on conventional electron spin resonance spectra of cholestane, fatty acid and alkylammonium-type spin probes has been studied in aqueous 1,2-dipalmitoyl-sn-phosphatidylcholine dispersions. The cooperative onset of the cholestane spin probe rotation about its long axis, with an effective correlation time of 2-3 ns, has been detected at a temperature coinciding with the calorimetric substransition, indicating onset of the host lipid rotational motion. The lipid rotation results in dissolution of the spin probe clusters in the host lipid. In the gel phase, the lateral distribution of impurity molecules is more isotropic than in the subgel phase. PMID- 2551737 TI - Purification of a DNA methyltransferase from Bacillus natto B3364. AB - An S-adenosyl-L-methionine:DNA-methyltransferase, termed M.BnaI, was purified from Bacillus natto B3364 strain by successive column chromatography. The molecular weight determined by gel filtration was 37 kDa for M.BnaI. Analysis of methyltransferase by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed correspondence of the M.BnaI activity with one protein band at a molecular weight of 35 kDa. Sequencing of pUC19 DNA methylated with M.BnaI showed the cytosine-5 methylation in the BnaI recognition sequence GGAT decreases CC at the position indicated by the arrow. PMID- 2551738 TI - Voltage-dependent transient calcium currents in freshly dissociated capillary endothelial cells. AB - Dissociated capillary endothelial cells display a voltage-dependent Ca current activating around the resting potential. The initial transient component of the current corresponds to a Ca channel of the T type. Some cells also display a plateau component corresponding to a distinct dihydropyridine-sensitive Ca channel. Depolarization induced by high external K+ elicits an increase in cytoplasmic Ca concentration. Confluent cells have been found to express the same Ca permeabilities. PMID- 2551739 TI - Thyroid hormone effect on rat heart mitochondrial proteins and affinity labeling with N-bromoacetyl-3,3',5-triiodo-L-thyronine. Lack of direct effect on the adenine nucleotide translocase. AB - N-bromoacetyl-3,3',5-tri[3'-125I]iodo-L-thyronine was used to label intact heart mitochondria from eu, hypo- and hyperthyroid rats in order to identify proteins involved in T3-regulated mitochondrial processes. The results show strong labeling, competed for by T3 and other analogues, of two proteins with a molecular mass of 48,000 and 49,200 Da. No labeling is seen of the adenine nucleotide translocase, a likely target, neither at 0 degree C, at room temperature, nor after preincubation with the substrates or specific inhibitors. No difference in labeling intensity or distribution is seen in mitochondria from eu-, hypo- or hyperthyroid rats, and the abundance of the adenine nucleotide translocase is unchanged, but five other proteins show differential abundance. PMID- 2551740 TI - Calcium channels in undifferentiated PC12 rat pheochromocytoma cells. AB - Undifferentiated rat pheochromocytoma PC12 cells were voltage clamped using the whole cell technique. After blockade of outward currents, calcium currents were elicited from -40 and -100 mV. A subpopulation of cells displayed only one current component activated at -10 mV and slowly decaying. In other cells this current coexisted with a component activated around -40 mV and decaying with a faster time constant. We conclude that undifferentiated PC12 cells can express two types of calcium channels, L (long-lasting) and N (neuronal)-type channels. PMID- 2551741 TI - Purification of 25-hydroxyvitamin D3 24-hydroxylase from rat kidney mitochondria. AB - 25-Hydroxyvitamin D3 24-hydroxylase was purified to an electrophoretically homogeneous state (Mr = 53,000) from kidney mitochondria of female rats treated with vitamin D3. PMID- 2551742 TI - Different effects of sphingosine, R59022 and anionic amphiphiles on two diacylglycerol kinase isozymes purified from porcine thymus cytosol. AB - Porcine thymus cytosol contains two immunologically distinct forms of diacylglycerol kinase (DGK) [Yamada, K. and Kanoh, H. (1988) Biochem. J. 255, 601 608]. These 2 DGK species, having apparent molecular masses of 80 and 150 kDa, were purified from the thymus cytosol. Upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the 150-kDa DGK gave 2 polypeptide bands of 50 and 75 kDa, whereas the 80-kDa DGK yielded a single protein band. The 80-kDa DGK was markedly activated by 10-20 microM sphingosine as well as by the known anionic activators such as phosphatidylserine and deoxycholate. In contrast, the 150-kDa DGK was fully active in the absence of the anionic activators and was strongly inhibited by sphingosine (IC50, 20 microM). The putative DGK inhibitor R59022 inhibited the 80-kDa DGK (IC50, 10 microM), but had little effect on the 150-kDa form. It is therefore clear that in the thymus cytosol there are at least 2 DGK isozymes operating under different control mechanisms. PMID- 2551743 TI - Relationship of symptom duration and survival in patients with colorectal carcinoma. AB - An early diagnosis in asymptomatic patients and a prompt treatment lead to an improved survival rate in patients with carcinoma of the colon. Patients with a short symptomatic history of colon cancer do not have a better prognosis than patients with a long history. Between 1978 and 1984 a consecutive series of 571 patients with colorectal cancer were admitted to the First Department of Surgery of the University of Rome. All patients were classified into five groups according to the duration of specific intestinal symptoms. In Group 1 (51 cases) asymptomatic patients were included, or patients with no specific symptoms such as asthenia, anemia, occult fecal blood. In Group 2 there were 129 patients with intestinal symptoms of less than 3 months' duration before treatment. In Group 3 there were 192 patients with symptoms of between 4 and 6 months' duration; 151 patients with symptoms of between 6 and 12 months were included in Group 4, and finally 48 patients who presented with symptoms of more than 1 year were included in Group 5. No relationship was noted between tumor site and duration of symptoms. Similarly, no relationship was noted between the duration of intestinal symptoms and stage and tumor differentiation. On the other hand, asymptomatic patients showed a higher incidence of T1N0M0 stage tumor and a lower percentage of undifferentiated neoplasms. The resectability rate was 79% and it was significantly related to the absence of intestinal symptoms. Follow-up data were available in 454 patients (80%). The overall survival rate was 52.4%. In Group 1 through Group 5 the 5-year survival rate was: 83.7%, 50%, 50%, 46.3%, 46.9%. The results of our study indicate that patients admitted in asymptomatic phase presented less-advanced stage tumors and, thus, best survival rate. On the other hand, from our data the duration of intestinal symptoms is not related to the stage and prognosis of tumors. PMID- 2551744 TI - [Isolation and fractionation in percoll gradient of guinea pig adrenal cortex cells and their functional characteristics]. AB - Suspension of isolated adrenal cells from the guinea pig adrenals was prepared using two modifications of collagenase dispersion. The cells separated by percoll density gradient gave 4 bands. The fraction of adrenocorticocytes has buoyant density of 1.03-1.05 g/ml. The lower bands comprise blood cells with density of 1.07-1.10 g/ml. The aldosterone content in the adrenocorticocyte fraction is 1200 1300 pg/10(5) cells. Intensity of labelling of cellular proteins increases only in the adrenocorticocyte fraction in response to a rise in K+ concentration in incubation medium. The Scatchard plot method was used to characterize binding of 125I-ACTH to dispersed adrenocortical cells. Binding association constant (Ka) and binding capacity (Bmax) are 1.3 X 10(8) M-1 and 2.0 X 10(-9) mol/l, respectively. A relation between functional activity of adrenocortical cells, their ultrastructural features and density is discussed. PMID- 2551745 TI - [Increased plasma and urinary Na+-K+ ATPase inhibitory activity and elevated blood pressure in metoclopramide-treated rats]. AB - Our recent studies demonstrated that dopaminergic receptor may play an important role in controlling blood pressure and renal sodium handling. This study was designed to investigate whether endogenous Na+-K+ ATPase inhibitors are related to changes in blood pressure and sodium balance, caused by the administration of metoclopramide (MC), a dopamine (D2) receptor antagonist, and sodium loading. Young male Wistar rats (250 approximately 300 g) were raised under three different conditions, control (control group), 1% NaCl loading (NaCl group) and 1% NaCl plus 1.5 mg/kg/day of metoclopramide (MC group) for 7 days. Systolic blood pressure (SBP), urinary and plasma Na+-K+ ATPase inhibitory activity (ATPI), urinary sodium, potassium and catecholamine excretions were measured. SBP in the MC group elevated from 116 +/- 2 mmHg to 134 +/- 4 mmHg, but there were no changes in SBP in the control and NaCl groups. Urine volume and urinary sodium excretion significantly increased in not only the NaCl group but also in the MC group. Plasma ATPI in both the NaCl and MC groups was higher than in the control group (p less than 0.001), and the level of ATPI in the MC group was significantly higher than in the NaCl group (p less than 0.05). Urinary ATPI in the control group was not detected after 7 days, but that in the NaCl and MC groups was clearly increased. The excretions of urinary catecholamines remained unchanged in all groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551746 TI - [Effect of cyproterone acetate on aromatase activity in cultured human genital skin fibroblasts: intracellular control of aromatase activity]. AB - Cyproterone acetate(CA), a well-known competitive antiandrogen, has been used for the treatment of precocious puberty, prostatic adenocarcinoma, hirsutism and hypersexuality. However, there have been some reports of troublesome gynecomastia developing during the use of this drug. It was, therefore, of interest to investigate the effect of CA on peripheral aromatization, since it is the major source of circulating estrogens in men. Our recent studies of aromatase activity in human skin fibroblasts demonstrated that the skin is an important site of extraglandular aromatase activity in men and suggested that these cells might provide a valuable new system in which to study the enzyme. Estrogen formation was assayed by the [3H]H2O technique, after 3h incubation of the cells with androstenedione. The initial experiment was designed to test the effect of CA (10(-8) to 10(-5) M) on baseline aromatase activity during a 12h preincubation in the presence of fetal bovine serum (FBS). Baseline aromatase activity was not affected by the presence of CA, whereas medroxyprogesterone acetate, a similar synthetic progestogen, induced a 2-fold stimulation of aromatase activity at a concentration of 10(-5) M. In cells preincubated with dexamethasone (DEX) in the presence of FBS, aromatase activity was stimulated markedly. When the cells were preincubated in the medium containing FBS with DEX (2.5 X 10(-7) M) in the presence of CA (10(-7) to 10(-4) M), DEX-stimulated levels of aromatase activity were inhibited by CA in a dose-dependent fashion. A competitive binding assay using [3H]DEX, showed that CA was able to compete with DEX for glucocorticoid receptor and the relative binding affinity of CA was approximately 50 times less than DEX. This suggested that the inhibitory effect of CA was due to competition with DEX for receptor binding. Aromatase activity was also stimulated by (Bu)2cAMP (1mM) in the absence of FBS. The stimulatory effect of (Bu)2 cAMP was maximal after 12-24h of preincubation, and this level was maintained for 60h. Similar to the DEX stimulation, stimulation of aromatase activity by (Bu)2cAMP required both RNA and protein synthesis, since the stimulatory effect of (Bu)2cAMP was abolished by co-preincubation with cycloheximide or actinomycin D. When CA was present during either the 12h preincubation or assay incubation, no difference was found in the (Bu)2cAMP-stimulated levels of aromatase activity. On the other hand, the non-aromatizable androgen dihydrotestosterone (DHT) (10(-8) to 10(-6) M) inhibited the stimulation of aromatase activity by (Bu)2cAMP in a dose-dependent fashion.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2551747 TI - Leukocytes of rainbow trout (Oncorhynchus mykiss) pronephros: cell types producing superoxide anion. AB - Fish pronephric and blood leukocytes yield a chemiluminescent response (CL) when stimulated appropriately. This response reflects the production of highly reactive oxygen derivatives which contribute to oxygen-dependent killing of targets such as pathogens and parasites. From suspensions of pronephric cells of the Rainbow trout, Oncorhynchus mykiss, we have obtained populations enriched for CL-positive cells. Four bands of cells were obtained using continuous gradients generated with 60% Percoll. The leukocytes of band II showed a very strong PMA induced CL response, the magnitude of which was several times higher than that observed with equivalent numbers of cells form unseparated pronephric cell suspensions. Cells present in other bands were not significantly chemiluminescent. Flow cytometric analysis showed that band II contained large granular cells and small granular cells. Cytochemical analysis showed that this subpopulation was greatly enriched with neutrophils. Many band II cells adhere to glass whereas few band III cells do so. The glass-adherent cells loose their CL potential after a few days in vitro, whereas the nonadherent cells retain their CL responsiveness for at least a week in vitro. PMID- 2551749 TI - Enzymatic collagen degradation in the pregnant guinea pig cervix during physiological maturation of the cervix and after local application of prostaglandins. AB - The role of enzymatic collagen degradation in prostaglandin-induced and physiological cervical ripening was studied in guinea pigs. The cervices were removed from (a) 8 non-pregnant guinea pigs, (b) 8 animals at day 45 of pregnancy, (c) 14 pregnant animals of comparable gestational age which had either an intracervical application of 0.2 ml 5% tylose or 10 micrograms sulprostone gel, and (d) 8 guinea pigs at day 63 to 65 of pregnancy. Collagenase activity was assayed in a highly specific and sensitive system using native collagen type I as substrate. Protease activity was measured by the method of Green and Shaw. Collagen fragments were identified by SDS-polyacrylamide electrophoresis (SDS PAGE) of acetic-soluble fractions. Collagenase and protease activities were found in all extracts from the different groups. However, there were no differences in enzymatic activities between the non-pregnant, early-pregnant and late-pregnant cervical specimens. Prostaglandin pre-treatment of the cervix led to no significant increase in either collagenase or protease activity as compared to the control groups. The absence of typical collagen degradation products in the SDS-PAGE suggested that no significant collagen breakdown had taken place. In contrast to previously published literature, we conclude that enzymatic collagen degradation is unlikely to be a key factor in prostaglandin-induced and physiological cervical ripening. PMID- 2551748 TI - Identification and production of monoclonal antibodies against a discriminating protein molecule between B and D variants of encephalomyocarditis virus. AB - The protein profiles of two EMC virus variants B and D in the infected L929 fibroblast lysates were examined using one- and two-dimensional gel electrophoresis. Both variants yielded a protein molecule of similar molecular weight but differing in its isoelectric point (pI). The B variant lysate yielded a molecule with pI congruent to 7.5 whereas the same molecule from the D lysate focused at pI congruent to 5.2. A monoclonal antibody (MCA) produced against pI congruent to 5.2 fraction of the D variant successfully detected viral antigens in the D variant infected fibroblasts with only background cross reactivity with the B variant infected cells. This MCA also detected D viral antigen(s) in organ sections obtained from D but not from B variant infected mice. This study therefore suggests a clear shift in the pI of a 23 kD protein molecule serving as a possible discriminating antigen between the B and D variants of EMC virus. PMID- 2551750 TI - Structural analogies between different redox enzymes inserted into biological membranes. PMID- 2551751 TI - Altered expression and distribution of the cytoskeletal-associated p35 protein in NIH 3T3 cells transformed with the Harvey sarcoma virus v-ras oncogene. AB - 1. Cytoskeletal events associated with retroviral oncogene (v-ras)-mediated transformation were studied in NIH 3T3 fibroblasts and their v-ras-transfected counterparts (3T3/H-1 cells). 2. Abnormal microfilament networks seen in 3T3/H-1 cells reflected significant decreases (approximately 90%) in two cytoskeletal associated proteins (tropomyosin-1, p35). Neither actin content nor actin mRNA levels were altered, however, v-ras transfectants. 3. p35 mRNA activity in both NIH 3T3 and 3T3/H-1 cells was similar although differential compartmentalization of p35 to the detergent-resistant cytoskeletal fraction was evident only in normal fibroblasts. 4. Proper cytoskeletal organization may be a factor in the regulation of p35 mRNA translation in situ or influence the stability of p35 independent of translational rate. PMID- 2551752 TI - Binding characteristics of high density lipoprotein subclasses to porcine liver, adrenal and skeletal muscle plasma membranes. AB - 1. We compared binding characteristics of 125I-labeled high density lipoprotein (HDL) subclasses to porcine liver, adrenal and skeletal muscle plasma membranes. 2. HDL subclasses were discriminated by their buoyant densities (HDL2 and HDL3) or by their apolipoprotein (apo) content (Lp-AI (particles containing apoA-I but no apoA-II) and LpA-I/A-II (particles containing both apoA-I and apoA-II)). 3. HDL2 and HDL3 showed saturable binding to the three types of membrane preparations. 4. No differences were found in the Kds within one HDL subclass. 5. Kds and maximal binding of HDL2 were lower than these of HDL3. Unlabeled HDL2 and HDL3, but not LDL, effectively displaced 125I-HDL2 and 125I-HDL3. 6. Binding of HDL was independent of the concentration of NaCl and did not require calcium. 7. These results suggest a process mediated by a single specific receptor in porcine liver, adrenal and skeletal muscle plasma membranes. 8. We also studied binding characteristics of HDL subclasses Lp-AI and LpA-I/A-II to porcine liver membranes. LpA-I showed the highest Kd and maximal binding. 9. All types of HDL subclasses studied (i.e. HDL2, HDL3, LpA-I and LpA-I/A-II) effectively competed for binding of both Lp-AI and LpA-I/A-II, suggesting that the HDL subclasses studied bind to the same receptor by their apoA-I moiety. PMID- 2551753 TI - Adenylate kinase of Mycobacterium marinum. Amino acid composition, secondary structure and other aspects. AB - 1. Adenylate kinase purified from Mycobacterium marinum shares homology with the enzyme from mammalian muscle and from other bacteria with respect to amino acid composition, total residues and Mr. 2. However, it differs from others in not containing a free sulfhydryl or disulfide group. 3. It is much more sensitive to inhibition by adenosine (5')pentaphospho(5')adenosine than is the enzyme from other sources. 4. The enzyme contains 24% alpha-helix, 30% beta-structure and 46% random coil. PMID- 2551754 TI - Ostrich fructose 1,6-bisphosphatase: distribution of activity and purification of the liver isoenzyme. AB - 1. Fructose 1,6-bisphosphatase was assayed in crude extracts of physiologically important organs and tissues in the ostrich. 2. Highest activity was found in liver and lowest in brain tissue. 3. No activity was detected in the heart, gizzard or adrenals. 4. The enzyme was purified in homogeneous, apparently undegraded form from liver utilizing Blue dextran-Sepharose affinity chromatography. 5. The enzyme is similar to mammalian fructose 1,6-bisphosphatase in many respects including its indispensability of Mg2+ for catalytic activity. 6. Relative molecular weight of the native enzyme and its subunit is about 150,000 and 35,000 respectively. 7. The amino acid composition of ostrich liver fructose 1,6-bisphosphatase is distinctly different from that of the chicken muscle enzyme, but compares favourably with the composition of the rabbit liver enzyme. 8. The purified enzyme is devoid of tryptophan. PMID- 2551755 TI - Steroidogenesis activator polypeptide (SAP) in the guinea pig adrenal cortex. AB - Steroidogenesis activator polypeptide (SAP), a cytosolic stimulator of cholesterol side-chain cleavage (cholesterol SCC) previously characterized in the rat, was isolated from guinea pig adrenal cortex. This factor exhibited behavior on reverse-phase high-performance liquid chromatography (HPLC) that was indistinguishable from authentic SAP and crossreacted fully in a SAP radioimmunoassay. In dexamethasone-suppressed guinea pigs neither the concentrations of immunoreactive adrenal SAP nor the levels of cholesterol SCC activity were significantly different between the outer zones (zonae glomerulosa and fasciculata) and the inner zone (zona reticularis). However, at 10 min after treatment of dexamethasone-suppressed animals with ACTH1-24, the outer zone content of SAP was increased 42-fold over unstimulated controls whereas inner zone SAP was elevated only 4-fold. At the same time, cholesterol SCC activity was increased 2-fold in the outer zones but unchanged in the inner zone. In addition to SAP itself, a crossreacting 82 kDa protein (p82)--similar to the putative SAP precursor identified in the rat--was detected on two-dimensional immunoblots of guinea pig whole adrenal homogenate. There were no significant differences in the protein concentrations of p82 or of cytochrome P-450scc between zones, either with or without ACTH treatment. We conclude that the widely reported contrast in corticosteroidogenic potential between the zona fasciculata and the zona reticularis of the guinea pig may reflect a differential capacity to generate SAP, and thus activate cholesterol SCC, in response to ACTH. PMID- 2551756 TI - Catechol estrogen formation in MCF-7 cell culture and effects of bromoestrogen inhibitors. AB - Agonist and antagonist activities have been reported for several catechol estrogens given exogenously. Since the metabolic clearance rate for catechol estrogens in the body is very rapid, catechol estrogens produced at other tissues will have minimal effect on breast tissue. Information of the extent of catechol estrogen formation within cells is critical in assessing the overall importance of these estrogen metabolites. Investigations of the conversion of estrogens to catechol estrogens were performed in the MCF-7 human mammary carcinoma cell culture system. Reverse-phase high-performance liquid chromatography (HPLC) analysis demonstrated that very little metabolism of estradiol occurs after 48 h, with only small amounts of estrone, 2-hydroxyestradiol, 2-hydroxyestrone, and estriol being observed. The total amount of 2-hydroxyestrogen products formed from 1 microM estradiol was 406.2 pmol (SD = 60.9) per 3 x 10(7) cells in 48 h. Similar results were obtained using the simpler radiometric assay for estrogen 2 hydroxylase, which measures the release of 3H2O from [2(-3)H]estradiol. The effects of inhibitors of estrogen 2-hydroxylase were also examined in MCF-7 cells. 2-Bromoestradiol, 4-bromoestradiol, and 2,4-dibromoestradiol effectively block estrogen 2-hydroxylase in a dose-dependent manner in MCF-7 cultures, with ED50 of approximately 1 microM for each inhibitor. Furthermore, these bromoestrogens bind poorly to estrogen receptors in MCF-7 cells and do not alter cell growth. Thus, in MCF-7 mammary cell cultures, metabolism of estradiol occurs to only a minor degree, and it is unlikely that the levels of catechol estrogens would reach physiologically relevant concentrations in the intact breast cancer cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551757 TI - Desensitization to follicle-stimulating hormone in cumulus cells is coincident with hormone induction of oocyte maturation in the rat follicle. AB - The objective of the present studies was to assess whether hormone induction of oocyte maturation in isolated intact follicles may be linked to desensitization of follicle-stimulating hormone (FSH) in the oocyte-cumulus complex (OCC). Incubation of follicles with chorionic gonadotropin (hCG), FSH or epidermal growth factor (EGF) produced a marked inhibition of FSH-dependent cyclic AMP accumulation in OCC with a time-course coincident with the onset of germinal vesicle breakdown (GVBD). These effects were evident within 3 h for both hCG and FSH, but with EGF a reduced response to FSH was seen within 1 h of treatment followed by an increase in GVBD. In contrast, no inhibition of cyclic AMP accumulation was seen in response to cholera toxin, forskolin or LH in OCC derived from follicles incubated with hCG for 3 h. The time-course for induction of oocyte maturation by incubation of the intact follicle with hCG was also coincident with production of prostaglandin (PG) F2 alpha, an indirect marker of cyclooxygenase induction. No effect on metabolic coupling between the oocyte and cumulus cells was seen until 9 h after hCG treatment. Retinoic acid caused a marked decrease in metabolic coupling between the oocyte and cumulus cells but inhibited oocyte maturation both in denuded oocytes and OCC. Since FSH desensitization in OCC, the resumption of meiosis, and production of arachidonic acid-derived products were coincident, it is suggested that abrogation of FSH action in cumulus cells by the ovulatory surge of gonadotropins may initiate oocyte maturation. PMID- 2551758 TI - Stimulation by atrial natriuretic factor of cyclic GMP production in cultured anterior and intermediate pituitary tissues: evidence for a major contribution of proliferating nonendocrine cells. AB - Primary cultures of anterior and intermediate pituitary tissues were monitored immunocytochemically for the presence of endocrine and nonendocrine cells and simultaneously tested for their ability to produce cyclic GMP in response to atrial natriuretic factor (ANF). Cells cultured for 3 days and 6 days, in which nonendocrine (vimentin-positive) cells were found to rapidly overgrow the endocrine cells, showed a dramatic elevation in cyclic GMP production stimulated by ANF, with maximum stimulation 300-700% that seen in 1-day cultured cells. Also, ANF-induced accumulation of cyclic GMP in an enriched population of vimentin-positive cells appeared to closely match that triggered in a 3-day culture of anterior pituitary cells, emphasizing the major role played by nonendocrine cells and their ability to synthesize cyclic GMP. In contrast, in the homogeneous population of tumor corticotrophs AtT-20, there was a close relationship between cyclic GMP formation and cell density. It thus appears that contamination of primary cultures of anterior and intermediate pituitary tissues by proliferating nonendocrine cells (mainly fibroblasts), in which ANF-induced accumulation of cyclic GMP may be confused with that of the very secretory cells, leads to overestimation and masking of guanylate cyclase activity of endocrine cells. PMID- 2551759 TI - Rhythmic oscillations of cytosolic free calcium in rat C-cells. AB - The relationship between stimulation of single C-cells (rMTC-6-23 cell line) with extracellular calcium, glucagon or 8-bromo-cAMP and fluctuations of intracellular free calcium concentration was studied. After pretreatment of rMTC cells with either 1 microM glucagon (30-60 min) or 1 mM 8-bromo-cAMP (5 min) [Ca2+]i started to oscillate when extracellular calcium was raised to 3 mM. These fluctuations in [Ca2+]i could be stopped by chelating the external calcium with EGTA or by adding calcium channel blockers. The voltage-dependent calcium channels in the plasma membrane seem to play a major role in maintaining the oscillations of [Ca2+]i. PMID- 2551760 TI - Mechanism of IGF-I-stimulated glucose transport in human adipocytes. Demonstration of specific IGF-I receptors not involved in stimulation of glucose transport. AB - We demonstrate the presence of specific insulinlike growth factor I (IGF-I) receptors in human adipocytes. Competition studies with 125I-labeled IGF-I and unlabeled IGF-I, IGF-II, and insulin showed the specificity of 125I-IGF-I binding to the IGF-I receptors in adipocytes, membranes, and partially purified detergent solubilized extracts. The monoclonal antibody to the IGF-I receptor (alpha-IR3) inhibits 125I-IGF-I binding and immunoprecipitates the IGF-I receptor. In addition, the alpha-subunit of IGF-I receptor is approximately 10,000 Mr larger than the alpha-subunit of insulin receptor, and IGF-I stimulates phosphorylation of the beta-subunit of the IGF-I receptor. IGF-I stimulates basal glucose transport in human adipocytes, but the concentrations of IGF-I required for half maximal and maximal stimulation of glucose transport are 800- and 1000-fold greater than that of insulin. The possibility of IGF-I stimulating glucose transport by interacting predominantly with insulin receptors is suggested by data showing that 1) IGF-I competes with insulin-binding sites, 2) there is a lack of an additive effect with IGF-I and insulin in stimulating glucose transport, 3) alpha-IR3, which specifically inhibits IGF-I binding, does not inhibit IGF-I or insulin-stimulated glucose transport, 4) insulin-receptor antibody MA-10 inhibits IGF-I and insulin-stimulated glucose transport, and 5) IGF-I stimulates insulin-receptor autophosphorylation, although its effect is markedly decreased compared with insulin. In summary, human adipocytes possess specific IGF-I receptors. However, IGF-I stimulates glucose transport predominantly by interacting with the insulin receptor. PMID- 2551761 TI - Prevalence of neuropathy in newly diagnosed NIDDM and nondiabetic control subjects. AB - We evaluated the prevalence of peripheral neuropathy by clinical and electrophysiological criteria and the prevalence of autonomic parasympathetic nerve dysfunction by heart-rate variation during deep breathing (expiration-to inspiration ratio [E:1]) in 132 newly diagnosed non-insulin-dependent diabetic (NIDDM) subjects aged 45-64 yr and 142 randomly selected nondiabetic control subjects. The relationship of nerve dysfunction to the degree of hyperglycemia and insulin-secretion capacity were also investigated. Single and scattered symptoms and signs of peripheral neuropathy were found in both diabetic and control subjects. Symptomatic polyneuropathy was found in 1.5% of diabetic subjects but none of the control subjects. Polyneuropathy defined by clinical signs was found in 2.3% of the diabetic subjects and 1.4% of the control subjects. No subjects with both symptoms and signs were seen. Nerve conduction velocities (NCVs) were significantly slower in diabetic than control subjects. Polyneuropathy according to electrophysiological criteria was found in 15.2% of diabetic subjects but was not found in any control subjects. Electromyographic abnormalities were more common in diabetic than control women, but not significant differences were found in men. The resting heart rate was higher in diabetic than control women, but no significant difference was found in men. The mean E:I was significantly lower in diabetic men and women than control men and women. An abnormally low E:I was found in 9.2% of the diabetic men, 3.3% of the control men, 3.3% of the diabetic women, and none of the control women. NCV parameters, but not E:I, were inversely correlated with fasting blood glucose and glycosylated hemoglobin levels. A positive correlation between NCV and fasting and postglucose serum insulin levels was found in both groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551762 TI - A common mechanism for activation of the Na+/H+ exchanger by different types of stimuli. AB - The mechanism of activation of Na+/H+ exchanger by various stimuli was studied in the human epidermoid carcinoma cell line A431 and in peripheral blood mononuclear cells (PBM). Intracellular pH (pHi) was measured by using the fluorescent dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein. Stimulation of A431 cells by epidermal growth factor (EGF), bradykinin (BK), phorbol-12-myristate 13-acetate (PMA), and osmotic shrinkage resulted in exchanger activation. In PBM, activation of Na+/H+ exchanger was induced by concanavalin A (Con A) and phytohemagglutinin (PHA), as well as PMA and osmotic shrinkage. Inhibition of protein kinase C inhibited only PMA-stimulated exchanger activation in both cell types. When osmotic shrinkage was applied after exposure of the cells to any agonist, augmentation of exchanger activation by osmotic stress was observed. These findings suggest that various stimuli activate Na+/H+ exchanger through different mechanisms. Kinetic analysis demonstrated that activation of the exchanger by any type of stimulus resulted in modification of the apparent affinities for intracellular H+ (H+i) and intracellular Na+ (Na+i) in opposite directions. While there is an increased apparent affinity for H+i, the apparent affinity for Na+i decreases. This finding suggests that in A431 cells this phenomenon serves as a common mechanism for activation of Na+/H+ exchanger by different stimuli. PMID- 2551763 TI - Central neurotensin inhibits gastric acid secretion: an adrenergic mechanism in rats. AB - Although parenteral neurotensin (NT) inhibits stimulated gastric acid secretion, published reports on the effect of centrally administered NT on gastric acid secretion are conflicting. This study provides evidence suggesting that, in chronic gastric fistula rats, intracerebroventricularly administered NT (15-60 micrograms) significantly reduces both basal and pentagastrin-, 2-deoxy-D-glucose , and carbachol-but not histamine-stimulated gastric acid secretion. Using radioimmunoassay, the concentration of plasma immunoreactive NT increased from 30 to 200 pg/ml at 30 and 60 min, respectively after a single intracerebroventricular (i.c.v.) administration of NT at a dose of 60 micrograms. These serum NT concentrations can be reproduced by a constant NT i.v. infusion at 2 micrograms/kg.h. This parenteral infusion dose does not inhibit acid secretion as does i.c.v. NT. Pretreatment with the i.c.v. dopamine-2 receptor antagonists haloperidol or domperidone totally abolishes the inhibitory effect of i.c.v. NT on pentagastrin-stimulated gastric acid secretion. In contrast, pretreatment with the specific dopamine-1 receptor antagonist SCH 23900 or the specific dopamine-2 receptor antagonist sulpiride does not affect i.c.v. NT-induced inhibition of pentagastrin-stimulated gastric acid secretion. Pretreatment (intracerebroventricularly) with the alpha-adrenergic antagonist phentolamine blocks the antisecretory effect of i.c.v. NT. Administration of 3.0 micrograms NT per side directly into nucleus accumbens (NACB), using a stereotaxic technique, significantly reduces basal gastric acid secretion. This effect of central NT is blocked by pretreatment with intra-NACB haloperidol (0.5 microgram per side). These findings suggest that NT acts centrally to inhibit gastric acid secretion, an effect that may occur within NACB and be mediated by central nervous system alpha-adrenergic receptor activation. PMID- 2551764 TI - Comparative study of Clostridium difficile toxin A and cholera toxin in rabbit ileum. AB - The purpose of this study was to compare the effects of Clostridium difficile toxin A and cholera toxin on fluid secretion, intestinal permeability, and arachidonate metabolites in rabbit ileum. Injection of 25 micrograms of either purified toxin into 10-cm ileal loops caused significant increases in fluid secretion and intestinal permeability to mannitol as well as release of prostaglandin E2 into the lumen. Toxin A, but not cholera toxin, caused a severe inflammatory reaction of the lamina propria and necrosis of enterocytes as well as increased release of leukotriene B4. The toxin A-mediated increases in prostaglandin E2 and leukotriene B4 could be blocked by prior instillation of 10 mg of 5-aminosalicylic acid into ileal loops. 5-Aminosalicylic acid also significantly diminished the expected increase in mannitol permeability after both toxins, but had no significant inhibitory effect on fluid secretion or, in the case of toxin A, intestinal inflammation. Our results indicate that C. difficile and cholera enterotoxins differ substantially in their effects on the rabbit intestine. Clostridium difficile toxin A, an inflammatory toxin, produces a striking infiltration of the lamina propria with neutrophils that is associated with increased release of leukotriene B4. In contrast, cholera toxin does not cause inflammation or leukotriene B4 release. Increased release of prostaglandin E2 occurs after exposure to both toxins and appears to be correlated with increased intestinal permeability. PMID- 2551765 TI - Ursodeoxycholic acid in primary biliary cirrhosis: results of a controlled double blind trial. AB - We studied the effect of ursodeoxycholic acid on 18 women and 2 men with primary biliary cirrhosis, mainly stages I and II. After a 3-mo observation period, patients were randomized to a 9-mo treatment period with ursodeoxycholic acid, 10 mg/kg.day, or placebo. Two patients on placebo left the study. In all patients on ursodeoxycholic acid, mean values of serum glutamate dehydrogenase, aspartate and alanine aminotransferases, alkaline phosphatase, and gamma-glutamyl transpeptidase fell significantly by 48%-79% after 18-24 wk; 7 of 10 showed a mean decrease of 35% in immunoglobulin M after 24 wk. Prothrombin time, serum bilirubin, albumin, the antipyrin breath test, and plasma disappearance of indocyanine green were normal initially and did not change. Total serum bile acid concentrations increased; ursodeoxycholic acid became the predominant bile acid. No significant improvement occurred in the placebo group. Hepatic histology improved in 6 patients of the ursodeoxycholic acid group but deteriorated in 4 patients receiving placebo. In studies with erythrocyte membranes, changes in electron spin resonance revealed that ursodeoxycholic acid was less toxic than chenodeoxycholic or deoxycholic acid, and coaddition of ursodeoxycholic acid prevented their toxic effect. PMID- 2551766 TI - Aflatoxin B1 formamidopyrimidine adducts in human hepatocarcinogenesis: a preliminary report. AB - The presence of aflatoxin-B1-formamidopyrimidine, a persistent aflatoxin deoxyribonucleic acid (DNA) adduct, was investigated in vivo by immunohistochemical analysis in 14 paired hepatocellular carcinoma and nontumorous human liver tissue sections using a monoclonal anti-aflatoxin-B1 formamidopyrimidine antibody. Nuclear and cytoplasmic accumulations of adducts were found in 4 of 14 nontumorous specimens but in none of 14 tumorous tissues and in none of three normal control livers. In vitro adduct formation and cellular DNA was investigated with a modified DNA immunoblot assay. These studies revealed (a) no significant difference in the amount of adduct formed by DNA samples with or without integrated hepatitis B virus DNA, (b) no difference in the amount of adduct formed with DNA from either tumorous or nontumorous tissues from a given individual, and (c) remarkable and reproducible differences in the capacity of DNA from different individuals to form in vitro adducts. Our DNA immunoblot assay will facilitate further studies on the molecular role of aflatoxin-B1-form-amidopyrimidine adducts in human hepatocarcinogenesis. PMID- 2551767 TI - High plasma pancreastatinlike immunoreactivity in a patient with malignant insulinoma. AB - High levels of pancreastatinlike immunoreactivity were detected in the plasma (2.9 pmol/ml, greater than 200-fold the normal level), pancreas (2.9 nmol/g wet wt, greater than 450-fold the normal level), and liver (1.6 nmol/g wet wt) of a patient with pancreatic insulinoma with metastasis to the liver by a sensitive and specific radioimmunoassay for human pancreastatin. Antiserum was produced against the C-terminal fragment of human pancreastatin-(24-52), which was synthesized according to the sequence of human chromogranin A corresponding to that of pancreastatin. With the antiserum, intense immunocytochemical staining was detected in the tumors. Sephadex G-50 gel filtration showed that the tumors and plasma contained two molecular forms of pancreastatinlike immunoreactivity--a molecular form coeluted with synthetic human pancreastatin-52 and a larger molecular form (Mr approximately 12,000-15,000). The smaller form eluted in the same position as synthetic human pancreastatin-52 on reverse-phase high performance liquid chromatography. PMID- 2551769 TI - Subcutaneous emphysema as a complication of endoscopic sphincterotomy of the ampulla of Vater. PMID- 2551768 TI - Synergism between vasoactive intestinal polypeptide and carbachol-mediated responses in a human mucus-secreting colonic epithelial cell line. PMID- 2551770 TI - Comparative study of the symbiotic plasmid DNA in free living bacteria and bacteroids of Rhizobium leguminosarum. AB - The symbiotic plasmid (pSym) DNA present in bacteroids of strain RCR1001 of Rhizobium leguminosarum biovar viceae has been compared qualitatively and quantitatively with that present in free living bacteria by hybridization experiments with appropriate probes. A decrease in the relative amount of pSym DNA was observed in bacteroids as compared to bacteria. No rearrangements of the symbiotically expressed pSym borne genes were detected in bacteroids. PMID- 2551771 TI - The effect of exogenously-supplied nucleosides and nucleotides and the involvement of adenosine 3':5'-cyclic monophosphate (cyclic AMP) in the yeast mycelium transition of Ceratocystis (= Ophiostoma) ulmi. AB - The yeast-mycelium transition of Ceratocystis (= Ophiostoma) ulmi (NRRL 6404) was induced by exogenously-supplied nucleosides and nucleotides in defined liquid media. During the yeast-mycelium transition, intracellular adenosine 3':5'-cyclic monophosphate (cAMP) levels increased and maximum levels coincided with maximum germination. This, coupled with findings that the cAMP phosphodiesterase inhibitors, theophylline and caffeine, also induced germination and elevated levels of intracellular cAMP, indicated the involvement of cAMP in the regulation of the yeast-mycelium transition. PMID- 2551772 TI - Calmodulin in heterocystous cyanobacteria: biochemical and immunological evidence. AB - The occurrence activity and localization of calmodulin in three heterocystous cyanobacteria of the genus Anabaena were studied. Boiled crude extracts caused a Ca2+-dependent stimulation of NAD kinase. Such a stimulation was blocked by EGTA and chlorpromazine, SDS-PAGE and Western blot analysis using antiserum against eukaryotic spinach calmodulin, revealed a polypeptide of about 17 kDa. Immunogold localization of calmodulin gave a dense gold label in both vegetative cells and heterocysts. The label was mainly confined to the centroplasm in vegetative cells, while it was evenly distributed in the cytoplasm of mature heterocysts. PMID- 2551773 TI - High yield synthesis of the bovine leukemia virus (BLV) p24 major internal protein in Saccharomyces cerevisiae. AB - Bovine leukemia virus (BLV) p24 gene was expressed in Saccharomyces cerevisiae under the control of the PHO5 (encoding repressible acid phosphatase, rAPase) promoter. Yeast cells were transformed by a yeast-E. coli shuttle vector carrying the PHO5 promoter, the p24 gene and the CYC1 transcription terminator. After low inorganic phosphate (Pi) induction of the PHO5 promoter, p24 accumulated in the producing cells up to a concentration representing 10% of total soluble proteins. The expression level of p24 gene was not increased by insertion of the positive regulatory gene PHO4 on the p24 expression vector. The p24 produced in this system and incubated in crude yeast extract showed a remarkably high resistance to proteolytic degradation, a feature that presumably correlates with the compact globular conformation of the protein combined to the stabilizing effect of the N terminal residue. PMID- 2551774 TI - Biochemical and immunological characterization of the bovine leukemia virus (BLV) envelope glycoprotein (gp51) produced in Saccharomyces cerevisiae. AB - The nucleotide sequence coding for bovine leukemia virus (BLV) envelope glycoprotein gp51 was inserted into a yeast-Escherichia coli shuttle vector carrying the promoter and secretion signal sequence of PHO5 (the yeast gene coding for repressible acid phosphatase) and the CYC1 transcriptional terminator. Yeast cells transformed by this construction synthesized gp51 after PHO5 induction by inorganic phosphate deprivation. The yeast-expressed gp51 was partially glycosylated into heterodisperse protein molecules ranging from 40 to 48 kDa. No gp51 was excreted in the culture medium. The amount of protein accumulated in yeast cells was estimated to reach 0.06% of soluble proteins. This modest level of expression seemed to be due to the toxicity of gp51 to the yeast cell. The yeast-expressed gp51 products were used in enzyme-linked immunosorbent assays for the detection of antibodies in sera from BLV-infected animals; they were also screened for the presence of well-defined biological epitopes. In both studies poor reactivity was observed. Rabbits immunized with the recombinant gp51 showed high antibody titers to native BLV gp51. However, these antibodies did not neutralize BLV in vitro. PMID- 2551775 TI - Cloning and expression of the major inner capsid protein of SA-11 simian rotavirus in Escherichia coli. AB - The major inner capsid protein (VP6) of SA-11 simian rotavirus has been expressed in Escherichia coli using a cloned cDNA derived from SA-11 double-stranded RNA segment 6. The cloned gene was fused to the N-terminal coding sequence of lacZ resulting in the synthesis of a 44-kDa protein. Several smaller polypeptides were also observed, resulting predominantly from transcription and translation within the gene 6 coding sequence. The recombinant VP6 proved to be antigenic by immunoblot analysis using polyclonal serum against SA-11 rotavirus and by Western blot analysis using monospecific serum derived from purified viral VP6. PMID- 2551776 TI - Purification of enzymatically active poliovirus proteinase 3C produced in Escherichia coli. AB - A viral protein 3C of the poliovirus (PV) Sabin 2 strain, a possible core region of the viral proteinase, was expressed in Escherichia coli using a recombinant DNA technology. The protein was recovered as a soluble protein from the insoluble protein fraction of the bacterial lysate, and was purified by a simple procedure with column chromatography. The viral capsid precursor P1 (1ABCD) of the PV Sabin 3 strain, which had been similarly produced in E. coli, was mixed with the purified or crude recombinant 3C. Immunoblotting assay with monoclonal antibodies specific to capsid proteins 1C (VP3) and 1D (VP1) of the PV Sabin 3 strain revealed that the in vitro reaction products contained 1C (VP3), 1D (VP1) and 1ABC (VP0-VP3). The data indicated that processing of the polyprotein P1 by the recombinant 3C proceeded properly in vitro, although an undigested product, 1ABC, is always detected in the reaction mixture. The results strongly suggest that, in addition to a protein 3CD, the 3C protein itself is also catalytically active in the processing of the viral capsid precursor polyprotein P1. PMID- 2551777 TI - Members of the RTVL-H family of human endogenous retrovirus-like elements are expressed in placenta. AB - A cDNA clone homologous to the RTVL-H family of human retrovirus-like elements was isolated from a human placenta cDNA library. The nucleotide sequence of the 1084-bp cDNA revealed an open reading frame (ORF) that may encode a 146 amino acid protein with significant homology to retroviral proteases. Downstream from the putative protease ORF a 3' long terminal repeat (LTR) containing U3 and R regions was found. The cDNA sequence ends in a poly(A) tail appropriately positioned downstream from a polyadenylation signal in the LTR. Northern-blot analysis showed that several distinct RTVL-H homologous transcripts are present in human placenta. We also show that repetitive RTVL-H homologous sequences are present in the genomes of both gorilla and African green monkey. PMID- 2551778 TI - Expression vector system based on the chicken beta-actin promoter directs efficient production of interleukin-5. AB - We examined the promoter activity of the 1.3-kb chicken beta-actin gene sequence located between the 5' flanking region and the proximal region of the second exon. This promoter region showed higher promoter activity than the simian virus 40 (SV40) early promoter or the Rous sarcoma virus (RSV) long terminal repeat (LTR) as assayed by transient lacZ gene expression in mouse L cells. Furthermore, replacement of the 3' splice sequence in this promoter by that derived from the rabbit beta-globin gene resulted in a approximately 2.5-fold enhancement in the synthesis of beta-galactosidase (beta Gal). Introduction of the SV40 origin of DNA replication (ori) into the vector carrying this hybrid promoter, which we designate the AG promoter, markedly enhanced the production of beta Gal in an SV40 T antigen-producing cell, BMT10. We have constructed a useful vector containing the strong AG promoter, several unique restriction sites, a SV40 polyadenylation signal and the SV40 ori for transient expression of cDNA in BMT10 or COS cells. We demonstrate the use of this vector for efficient production of interleukin-5 in BMT10 cells. PMID- 2551779 TI - Expression of human HPRT mRNA in brains of mice infected with a recombinant herpes simplex virus-1 vector. AB - Complete deficiency of the purine salvage enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) results in a devastating neurological disease, the Lesch-Nyhan syndrome. This disorder has been identified as a candidate for initial attempts at somatic cell gene therapy. We have previously reported the construction of a recombinant herpes simplex virus type 1 (HSV-1) vector containing human hprt cDNA sequences under the regulatory control of the viral thymidine kinase gene (tk) [Palella et al., Mol. Cell. Biol. 8 (1988) 457-460]. Infection of HPRT- cultured rat neuronal cells with these vectors resulted in transient expression of human hprt. In this paper, we report the expression of human hprt mRNA transcripts in the brains of mice infected in vivo with this vector by direct intracranial inoculation. Human hprt transcripts were distinguished from endogenous mouse transcripts by RNase A mapping using riboprobes transcribed from human hprt cDNA. These initial studies demonstrate the transfer and transcription of a human gene in brain cells by direct in vivo infection with recombinant HSV-1 vectors. PMID- 2551780 TI - Cloning and expression of mouse-brain calmodulin as an activator of Bordetella pertussis adenylate cyclase in Escherichia coli. AB - Cloning of higher eukaryotic genes has seldom been performed by complementation of a defective prokaryotic function. This is especially true in the case of functions that are normally absent from the prokaryotic host. We demonstrate here that it is possible to identify by complementation the cDNA from mouse brain, which encodes calmodulin (CaM) synthesis, in spite of the fact that the recipient strain, Escherichia coli, does not normally harbour a CaM function. A three component cloning procedure in which a gene product requiring CaM for activity, adenylate cyclase from the pathogen Bordetella pertussis, was used to screen a cDNA library for cAMP synthesis in E. coli. The nucleotide sequence of the corresponding cDNA is also reported. PMID- 2551781 TI - A cassette with seven unique restriction sites, including octanucleotide sequences: extension of multiple-cloning-site plasmids. AB - A 27-bp synthetic DNA cassette was constructed which contains the restriction sites of the two rare-cutter enzymes NotI and SfiI and, in an overlapping arrangement, those of five enzymes with 6-bp recognition sequences: ApaI, BalI, NdeI, SacII, XmaIII. The protruding termini of the fragment allow its insertion into any EcoRI-cut DNA creating a new EcoRI site at one side of the cassette only. This fragment was integrated into the pUC18-like multiple-cloning-site (MCS) plasmids pTZ18R and pTZ19R, producing a set of vectors which carry seven additional unique restriction sites (giving a total of 17) within their MCS. They still provide the capabilities of simple recombinant selection by blue/white coloured colonies, creation of single-stranded DNA in the presence of a helper phage, and in vitro transcription of cloned DNA using T7 RNA-polymerase. Plasmids with two copies of the DNA cassette inserted into their MCS were also constructed. Beside the advantages they provide in some cloning procedures, these latter plasmids, which carry a tandem repeat, are valuable sources of related 27 bp fragments, with features similar to the original but with different cloning termini. PMID- 2551782 TI - New derivatives of transposon Tn5 suitable for mobilization of replicons, generation of operon fusions and induction of genes in gram-negative bacteria. AB - Three types of new variants of the broad-host-range transposon Tn5 are described. (i) Tn5-mob derivatives with the new selective resistance (R) markers GmR, SpR and TcR facilitate the efficient mobilization of replicons within a wide range of Gram-negative bacteria. (ii) Promoter probe transposons carry the promoterless reporter genes lacZ, nptII, or luc, and NmR, GmR or TcR as selective markers. These transposons can be used to generate transcriptional fusions upon insertion, thus facilitating accurate determinations of gene expression. (iii) Tn5-P-out derivatives carry the npt- or tac-promoter reading out from the transposon, and TcR, NmR or GmR genes. These variants allow the constitutive expression of downstream genes. The new Tn5 variants are available on mobilizable Escherichia coli vectors suitable as suicidal carriers for transposon mutagenesis of non-E. coli recipients and some on a phage lambda mutant to be used for transposon mutagenesis in E. coli. PMID- 2551783 TI - The tubulin gene cluster of Trypanosoma brucei starts with an intact beta-gene and ends with a truncated beta-gene interrupted by a retrotransposon-like sequence. AB - In Trypanosoma brucei the genes coding for alpha- and beta-tubulin are arranged in a single cluster comprising about ten alternating alpha and beta genes. The cluster starts at its 5' end with an intact beta gene. In this area, a 1.1-kb segment upstream of the mini-exon addition site has been sequenced. Several potential transcription signals were seen. The 3' end of the cluster terminates in the middle of a beta gene, making it an inactive truncated pseudogene. The adjoining 1.7-kb sequence shows a high degree of similarity with a retrotransposon-like (RTnL) element previously characterized elsewhere in the trypanosome genome. This is the first report of an RTnL element interrupting a protein-coding gene of T. brucei. PMID- 2551784 TI - Efficient cloning of a mutant adenylate-kinase-encoding gene from Escherichia coli. AB - An optimized system has been developed for the transfer of a mutant gene from the Escherichia coli chromosome to a plasmid carrying the wild type (wt) allele. The wt allele was first cloned into a low-copy-number, self-transmissible plasmid with a single EcoRI, HindIII, and BamHI site. The plasmid was then transferred to a mutant strain that had been previously transformed with a high-copy-number plasmid carrying the recA+ gene to allow efficient homologous recombination. A 15% frequency of homogenotization was obtained during cloning of an adk gene that encodes a temperature-sensitive adenylate kinase (AK). The mutant AK had decreased mobility on sodium dodecyl sulfate-polyacrylamide gels compared with the wt enzyme. This was due to a point mutation that changed leucine-107 in the wt enzyme to glutamine-107 in the mutant enzyme as determined by nucleotide sequencing. PMID- 2551785 TI - Nucleotide sequence and analysis of the Vibrio alginolyticus sucrase gene (scrB). AB - The nucleotide sequence of a 2.119-kb DNA fragment containing the Vibrio alginolyticus sucrase gene (scrB) was determined. The complete sequence (484 aa residues) of the sucrase was deduced and homology was detected between the sucrase enzymes from V. alginolyticus and the Gram-positive bacteria Bacillus subtilis and Streptococcus mutans. In Escherichia coli cells the cloned V. alginolyticus sucrase is translocated to the periplasm. Transposon phoA mutagenesis experiments strongly suggested that V. alginolyticus sucrase in E. coli is not exported across the cytoplasmic membrane by means of a typical signal sequence. PMID- 2551786 TI - Detection and molecular cloning of highly repeated DNA in the sea cucumber sperm. AB - A highly reiterated sequence in the sperm DNA of the echinoderm Holothuria tubulosa has been isolated by digestion with EcoRI, and cloned in the phagemid Bluescript. The monomeric unit has a repeat length of 391 bp and is arranged in tandem. The uncloned genomic monomer as well as two independent cloned fragments have been sequenced. The repeated element constitutes about 1.8% of total Holothuria DNA which corresponds to a repetition frequency of about 1.4 x 10(5) copies per haploid complement. The repetitive sequence has a high A + T content (66.8%) characterized by scattered tracts of A and T residues with no apparent internal sub-repeats, although several inverted and direct repeats are present. Heterogeneity between monomers derived from individual clones is low, whereas sequence similarity to known repetitive elements appears to be negligible. PMID- 2551787 TI - Alpha- and beta-adrenergic receptors of noradrenergic innervation modulate the lower limits of cerebral and cerebellar blood flow autoregulation in spontaneously hypertensive rats. AB - The effects of alpha- and beta-adrenergic receptors on the lower limits of cerebral (CBF) and cerebellar blood flow (CeBF) autoregulation were examined in spontaneously hypertensive rats. CBF and CeBF were measured during hemorrhagic hypotension in rats treated with adrenergic blockers, phenoxybenzamine (PBZ) or propranolol (PPL), using a hydrogen clearance method. The lower limits for CBF beyond which blood flow was decreased steeply were 72% of resting values in the control, 44 in the PBZ and 80 in the PPL group. Similar tendency was also observed for CeBF. In the control group, the reducing rates of mean arterial pressure (MAP) to decrease CBF by 15 and 30% of the resting values were 33% (58 mm Hg) of the resting MAP and 46% (80 mm Hg), respectively, and those to reduce CeBF to the same extent were 35% (61 mm Hg) and 52% (92 mm Hg), respectively. In the PBZ group, the respective reducing rates of MAP were 52% (71 mm Hg) and 65% (88 mm Hg), respectively, for CBF and 50% (68 mm Hg) and 65% (88 mm Hg), respectively, for CeBF. In contrast, in the PPL group, those rates were 22% (39 mm Hg) and 32% (56 mm Hg), respectively, for CBF, and 23% (40 mm Hg) and 30% (52 mm Hg), respectively, for CeBF, being significantly smaller than those in the control and the PBZ groups (p less than 0.01, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551788 TI - [Hygienic evaluation of hydrophobic turgid Perlite and the materials of powder composition used in water supply systems]. AB - Sanitary, hygienic, toxicologic, pathomorphologic and genetic studies revealed that hydrophobic pearlites g A-1 and g A-2 and the material from powder composition of the brand of zh g p1, 5 d 2.5 had no unfavourable effect on physicochemical water characteristics, warm-blooded animals and poorly organized organisms. Hydrophobic pearlites were recommended for application in water reservoirs for the removal of petroleum films and the material from powder composition of the above brand for manufacturing cocks, sleeves, showers and other devices for water supply systems used for economic and drinking purposes. PMID- 2551789 TI - [Behavior of enteroviruses in the soil]. PMID- 2551790 TI - [Photosensitive paper as an indicator of the virucidal effect of bactericidal lamps]. PMID- 2551791 TI - Spin trapping evidence for radical generation by isolated hearts and cultured heart cells. AB - The spin trap 5,5-dimethyl-1-pyrroline-1-oxide (DMPO) has been applied to monitor the generation of free oxy-radicals in samples derived from isolated hearts and heart cells. .OH was trapped in the effluent of isolated hearts in the early phase of reperfusion following an ischemia time of only 10 min. Radical detection was possible even when the cardioactive DMPO was added to the effluent after draining off the heart, demonstrating that the short-lived .OH was generated by components released from the affected heart. These results support the hypothesis that radicals are of relevance for reperfusion injury. By omitting antioxidants commonly used for incubation media of cultured cells, it was possible for the first time to demonstrate the formation of .OH in the incubation solution of cardiac cells. PMID- 2551792 TI - A study on spin-labelled oligonucleotide synthesis and its electron spin resonance behavior in solution. AB - An oligonucleotide spin-labelled with 4-amino-2,2,6,6-tetramethylpiperidine-N oxyl (4-amino-TEMPO) at the internucleotide bond (d-Tp(L)TpTpTpT) prepared by oxidation of the pentanucleotide containing the H-phosphonate diester (d Tp(H)TpTpTpT) in the presence of 4-amino-TEMPO, was separated and identified by high-performance, reverse-phase liquid chromatography combined with detection by electron spin resonance spectroscopy. This spin-labelled oligonucleotide produced a triplet with the slightly broadened M1 = -1 ESR component, while a triplet with almost equal intensities was obtained from the spin-label. The M1 = -1 component from the labelled oligonucleotide was further broadened in the presence of poly(A) which forms a complementary double strand with this molecule. PMID- 2551793 TI - Alzheimer's disease: a pathogenic role for aluminosilicate-induced phagocytic free radicals. AB - The occurrence of aluminosilicate deposits within the cerebral plaques in Alzheimer's senile dementia sufferers has prompted further consideration of the possible role of such materials in the aetiology and pathogenesis of the disease. We have monitored the ability of various natural and synthetic model aluminosilicate particulates of differing morphological and chemical composition to stimulate the generation of phagocyte-derived free radical reactive oxygen metabolites (ROM) using an in vitro chemiluminescent technique on purified human blood-derived polymorphonuclear leukocytes (PMN). The results indicate that an enhanced chemiluminescent response is produced by calcium-bearing fibriform particulates. It is proposed that an analogous in vivo particle-induced and phagocyte-mediated oxidative stress could provide a potential pathogenic mechanism in the development of Alzheimer's disease. PMID- 2551794 TI - Reaction of hydroxyl radicals with the ferrous and ferric iron chelates of diethylenetriamine-N,N,N',N",N"- pentaacetate. AB - Reactions of the hydroxyl radical with ferrous and ferric diethylenetriamine N,N,N',N",N"-pentaacetate (DTPA) complexes in neutral aqueous solution were studied using a 60Co-gamma radiolysis source for radical production. Hydroxyl radicals oxidize the ferrous chelate of DTPA to the ferric chelate, as determined by changes in the optical absorbance. In the presence of equimolar concentrations of ferric and ferrous chelates, HO. is "trapped" by Fe(III)DTPA2- and yields of Fe(III)DTPA2- are substantially reduced. Both tert-butyl alcohol and methanol react efficiently with hydroxyl radicals. tert-Butyl alcohol produces unreactive radicals, and is thus an effective scavenger. However, the reducing hydroxymethyl radical produced upon reaction of HO. with methanol effectively reduces Fe(III)DTPA2- present in the system, so that solutions initially containing the more absorbing Fe(III)DTPA2- are bleached. These results with alcoholic hydroxyl radical scavengers illustrate possible complications from scavengers such as methanol which produce reactive products which may themselves stimulate further reaction. PMID- 2551795 TI - Ovarian malignant luteinized thecoma--an unusual tumor in an adolescent. AB - Malignant luteinized thecoma is an extremely rare ovarian tumor. The fifth reported case of this tumor presenting in a 13-year-old epileptic child on anticonvulsant therapy with a rapidly enlarging pelvic-abdominal mass is discussed. At laparotomy, bilateral ovarian involvement, massive ascites, and omental and bowel serosal metastases were present. The pathological diagnosis was difficult with initial diagnoses of ovarian edema, endodermal sinus tumor, and ovarian fibrosarcoma. The final diagnosis of malignant luteinized thecoma was made by recognizing the presence of lutein cells with foamy vacuolated cytoplasm and intracytoplasmic staining for oil red O. Electron microscopy also demonstrated vacuolated cells with multiple fat globules. After surgical debulking the patient was treated with combination chemotherapy with complete response. The similarities of this case to three previously reported bilateral thecomas presenting in children on anticonvulsant therapy is discussed. PMID- 2551796 TI - Gamma-aminobutyric acid, its related enzymes and receptor-binding sites in the human ovary and fallopian tube. AB - The concentration of gamma-aminobutyric acid (GABA), the activities of related enzymes, i.e. glutamate decarboxylase and GABA transaminase, as well as the level of specific GABA binding sites were determined in ovaries and fallopian tubes obtained surgically from 31 women. None of the biochemical parameters examined showed a correlation with the age and hormonal background (serum estradiol and progesterone levels) of the patients. The respective ovarian and tubal values did not differ significantly in groups operated on because of uterine myoma and carcinoma. In organs from pregnant women, however, most GABAergic markers altered significantly. These findings indicate some gestation-related role for the ovarian and tubal GABA systems in humans. PMID- 2551797 TI - Congenital non-spherocytic haemolytic anaemias. AB - Congenital non-spherocytic haemolytic anaemias are a heterogeneous group of disorders caused by an ineffective normal-type enzyme or, more commonly, by a structurally abnormal and ineffective mutant enzyme. The first part of this paper deals with the laboratory diagnosis of these diseases, which is based on a three step approach: demonstration of the haemolytic nature of the disease, demonstration that the cause of haemolysis is intracorpuscular and demonstration of the presence of an absolute or relative enzyme deficiency. Moreover, the possible causes of false negative results in standard enzyme assays are briefly commented. The second part focuses on the main clinical, laboratory and biochemical features of the most common enzyme deficiencies of both the Embden Meyerhof pathway and purine and pyrimidine metabolism, with particular mention of the cases identified in Italy. PMID- 2551798 TI - Treatment of HIV-associated thrombocytopenia. PMID- 2551799 TI - Tubuloreticular inclusions and viral particles in a case of hairy cell leukemia. PMID- 2551800 TI - [Cough associated with captopril and enalapril]. AB - Persistent dry cough is not a known adverse reaction of captopril and enalapril. We present 5 patients with persistent dry cough severe enough to warrant withdrawal of the drug, which resulted in rapid and complete recovery. Challenge with the drugs induced recurrence of cough. The pathogenesis of the reaction is unknown, but possible mediators include bradykinin and prostaglandins. PMID- 2551801 TI - [General pharmacological studies on 5,8,11,14,17-eicosapentaenoic acid ethyl ester (EPA-E)]. AB - EPA-E, even at 3,000 mg/kg, p.o., did not affect the general behaviors, spontaneous locomotor activities, pentobarbital hypnosis and body temperature; and it did not elicit anticonvulsant, analgesic and muscle relaxant actions. It had no influence on spontaneous EEG activities, even at 3,000 mg/kg, i.d. EPA-E at concentrations up to 10(-4) M, did not affect the tonus or agonist-induced contraction of the isolated ileum, trachea, fundus and vas deferens. EPA-E had no influence on the spontaneous movement of isolated ileum or uterus. EPA-E did not affect the nictitating membrane contraction and intestinal propulsive motility, and it did not damage gastric mucosa nor elicit antiulcer action. EPA-E at 1,000 mg/kg were without effect on gastric secretory volume (SV), total acidity (TA) and pepsin activities (PA). However, EPA-E at 3,000 mg/kg significantly decreased SV and TA without significantly decreasing PA. EPA-E caused no changes in the respiration, blood pressure, heart rate and ECG at the doses up to 3,000 mg/kg; and it did not affect the heart rate and contractile force on the isolated atria at concentrations up to 10(-4) M. The intracutaneous injection of 2.0% EPA-E produced neither anesthetic nor irritative action. EPA-E did not elicit hemolytic action at 10(-4) M. EPA-E, even at 3,000 mg/kg, did not affect the neuro-muscular transmission, urine volume, urinary excretion of electrolytes and carrageenin edema. These results suggested that EPA-E has no noticeable effects on the central nervous, autonomic nervous, respiratory and cardiovascular systems and so on. PMID- 2551802 TI - [Structure and function of gamma-aminobutyric acid (GABA) receptor: current state and prospectives]. AB - The gamma-aminobutyric acid (GABA) receptor has been classified into two receptor subtypes (GABAA and GABAB receptors) based on their pharmacological properties. The GABAA receptor in the central nervous system (CNS) has been found to be coupled structurally as well as functionally with the benzodiazepine receptor and Cl- channel. Purified GABAA receptor from bovine brain consisted of both alpha and beta subunits. The complementary DNAs encoding the GABAA receptor alpha and beta subunits have been cloned; and from their elucidated nucleotide sequences, the amino acid sequences of the subunits were deduced. The structure of both subunits, having four putative membrane domains, has been found to be similar to other ligand-gated receptors such as the nicotinic acetylcholine receptor alpha subunit and glycine receptor 48K subunit. Therefore, it has been suggested that these ligand-gated receptors comprise a superfamily. In addition, the presence of similarities in the nucleotide and deduced amino acid sequences of human brain GABAA receptor with those of bovine brain has been noted. On the other hand, the GABAB receptor, which is insensitive to bicuculline but sensitive to baclofen, has been found to be pharmacologically distinct from the GABAA receptor. The GABAB receptor in the brain has been found to be coupled with GTP-binding protein and generates the inhibitory transmission coupled with various intracellular effector systems such as adenylate cyclase and phosphoinositides turnover. The exact structure and function of the GABAB receptor in the CNS, however, remain to be clarified in future studies. PMID- 2551803 TI - Phospholipase C-mediated intestinal mucosal damage is ameliorated by quinacrine. AB - Phospholipase C from Clostridium perfringens, when injected into a closed loop of the rat small intestine in vivo, caused an increase in the activity of intraluminal N-acetyl-beta-glucosaminidase and mucosal permeability to sodium fluorescein, indicating damage to the mucosa. Phospholipase C also caused an influx of granulocytes (neutrophils) into the mucosa, as shown by the myeloperoxidase activity--a granulocyte neutrophil marker, and increased localized lipid peroxidation. Pretreatment of animals with quinacrine, a known inhibitor of phospholipase A2, prevented the increases in the luminal N-acetyl beta-glucosaminidase activity, mucosal permeability, malondialdehyde and myeloperoxidase activity after deposition of phospholipase C in the gut lumen. It is concluded that phospholipase C might impair the function of the mucosal barrier and increase the permeability of the gut to undesirable molecules and pathogens. Part of its action may be mediated via phospholipase A2 activation since pretreatment with quinacrine afforded protection. PMID- 2551804 TI - Effects of dietary retinyl acetate on the promotion of hepatic enzyme-altered foci by polybrominated biphenyls in initiated rats. AB - Vitamin A inhibits the development of some chemically-induced tumours. Since polybrominated biphenyls (PBBs) are hepatic tumour promoters and they affect vitamin A homeostasis in rats, we put forward the hypothesis that dietary levels of vitamin A would influence tumour promotion by PBBs. In the study described here, female Sprague-Dawley rats were initiated on day 1 by ip administration of diethylnitrosamine. On day 7 after initiation, the rats were fed a vitamin A deficient basal diet that was supplemented with either 2000 IU (low-vitamin A) or 200,000 IU (high-vitamin A) retinyl acetate/kg feed. From day 30 after initiation until the end of the study the following PBBs were added to the diets: Firemaster BP-6 (10 ppm), 2,4,5,2',4',5'-hexabromobiphenyl (10 ppm) or 3,4,5,3',4',5' hexabromobiphenyl (1 ppm). The control animals received low- or high-vitamin A diets containing no PBBs. On day 180, the rats were necropsied, sections of various tissues were stained for histopathological examination and an evaluation of hepatic enzyme-altered foci was performed. Numbers of gamma-glutamyl transpeptidase-positive foci/cm3 liver and the mean volumes of these foci were lower in the high-vitamin A groups than those in the corresponding low-vitamin A groups, but these differences were not significant. The percentage of the liver volume occupied by foci was significantly greater in the low-vitamin A with 345 HBB group than in the corresponding high-vitamin A group. Thus, high dietary levels of vitamin A had some inhibitory effect on the promotion of hepatic altered foci by 345-HBB in initiated rats. PMID- 2551805 TI - [Modification of blood coagulation by antihypertensive therapy? Effect of enalapril and hydrochlorothiazide on blood coagulation parameters in patients with essential hypertension]. AB - An increased tendency of the blood to clot in patients with arterial hypertension is not desirable, since cardiovascular and cerebrovascular sequelae may be aggravated. Recently, activation of coagulation under ACE inhibition with captopril has been described. In an open, randomized study, we compared the influence of enalapril on various coagulation parameters with that of hydrochlorothiazide. In contrast to the latter, we detected no unfavorable influence of enalapril on the coagulation factors investigated. A transient increase in fibrin monomeres was observed in one patient each on enalapril and hydrochlorothiazide. PMID- 2551806 TI - [Differential diagnosis of epilepsies in early childhood]. AB - The last twenty years have seen substantial advances in knowledge concerning the diagnosis as well as the long-term evolution of epilepsies in infants and children. There is, however, no general agreement among epileptologists on the framework of epileptic syndroms in infancy and childhood. The main problems are the lack of a uniform terminology and the fact that there is little unanimity concerning definitions and what may be included under individual syndromic rubrics. In consequence the nosological limits between the various syndromes described are not distinct and they very often represent a heterogenous group of disorders and not a single entity. The present paper reviews the literature recently published and discusses diagnostic difficulties and nosological problems. In addition the attempt has been made to elaborate a "multi dimensional" classification of syndromes, based on clinical (using the terminology of the international classification) and electroencephalographic criteria, but also taking into consideration aetiological and evolution data. Results show that it was possible to differentiate within a cohort of 77 infants aged 0-24 months (suffering from various types of epileptic disorders) several distinct and homogenous nosological entities and syndromes. As it is well known today, that the type of epilepsy which occurs in a child as well as the prognosis of the disease represent a confluence of age, heredity and structural brain abnormality, it seems apparent that adequate classification systems have to take into consideration the complexity of various factors "triggering" the onset and influencing the course of the disease, even when a number of cases remains outside resp. "borderline" such a syndromic classification. PMID- 2551807 TI - Posthospital convalescence and return to work. PMID- 2551808 TI - Column perifusion system for steroidogenesis in isolated rat adrenal cells. AB - A perifusion system using a plastic column into which isolated rat adrenal cells had been installed was attempted. After ACTH or cAMP was administered to the column, the corticosterone concentration in the eluate was determined. ACTH in 10(-13) and 10(-12) M did not promote corticosterone production, whereas 10(-11) and 10(-10) M showed a dose dependent production of corticosterone. By iterative infusion of 10(-11) or 10(-9) M of ACTH, very clear responses to restimulation of ACTH were noted. Following the administrations of 10(-3) or 10(-2) M of dibutyryl adenosine 3',5'-cyclic monophosphate (dbcAMP), the production of corticosterone increased dose-dependently. These results suggest that this perifusion system is effective for examining the effects of ACTH or cAMP on steroidogenesis of cells. PMID- 2551809 TI - Stimulation of the chlorinating activity of human myeloperoxidase by thyroid hormones and analogues. AB - Thyroxine and triiodothyronine concentrations of 50 nM or lower can stimulate the chlorinating activity of the myeloperoxidase-H2O2-Cl- antimicrobial system in vitro. The initial rates of the chlorinating reaction with monochlorodimedone were similar for both thyroid hormones. Maximum stimulation occurred around pH 6 and a linear relationship exists between stimulation and thyroxine concentrations up to at least 1 microM. Of the various thyroxine analogues tested, stimulation was in the order: T4 (or T3) greater than triiodothyropropionic acid greater than 3,5-T2. Diiodotyrosine did not have any significant stimulatory effect. The oxidised product of the phenolic ring of T4, presumably a hydroxyquinone, may act as an additional electron carrier and thereby facilitates redox reactions. PMID- 2551810 TI - Normoreninemic hypoaldosteronism in a case of isolated ACTH deficiency. AB - This paper documents the rare and hitherto unreported association between isolated ACTH deficiency and normoreninemic hypoaldosteronism in a 63-year-old woman. Baseline plasma aldosterone and 18-hydroxycorticosterone were extremely low. Both steroids did not respond to exogenous angiotensin II infusion, whereas they were increased in parallel to ACTH stimulation. Thus, acquired dysfunction or congenital dysgenesis of the zona glomerulosa was suspected. The upright posture-furosemide test showed a subnormal but definite plasma aldosterone response coupled with a normal increase in plasma renin activity, indicating that there may be a yet unidentified mechanism(s) underlying the postural increase of aldosterone. PMID- 2551811 TI - GABAergic drugs and lordosis behavior in the female rat. AB - Agents modifying GABAergic neurotransmission were administered to ovariectomized rats treated with different doses of estradiol benzoate (EB) + progesterone (P) or with EB alone. Hormone treatments were designed to induce an intermediate level of receptivity in order to be able to observe both stimulatory and inhibitory effects on lordosis behavior. Both the GABAA receptor agonist THIP and the GABAB receptor agonist baclofen inhibited lordosis behavior at doses from 20 and 5 mg/kg, respectively. The GABA transaminase inhibitor gamma-acetylen GABA (GAG) and the GABA agonist 3-aminopropanesulfonic acid had no effects, even when high doses were administered. The GABAA receptor antagonist bicuculline had no effect by itself nor did it block the effects of THIP. It is therefore suggested that the GABAA receptor is of slight importance in the control of lordosis behavior. No evidence could be found supporting the hypothesis that an interaction between P and GABA is important for hormone-induced receptivity. It does not appear likely that motor disturbances are responsible for the inhibitory effects of baclofen and THIP. The exact mechanism by which these drugs inhibit lordosis behavior is not clear at present. PMID- 2551812 TI - [In vitro assessment of candidacidal activity of human leukocyte myeloperoxidase preparation]. AB - Myeloperoxidase (MPO), present in the azurophilic granules of human polymorphonuclear neutrophils, is important in the oxygen-dependent microbicidal activity of neutrophils. The purpose of our study was to investigate the therapeutic potency of the MPO preparations. This paper is to present our primary work on MPO isolation and its microbicidal activity assay. White blood cells, isolated freshly from normal donors, were lysed with cetyltrimehylammonium bromide to liberate myeloperoxidase. The enzyme preparation was partially purified by 50% (NH4)2SO4 precipitation followed by 65% (NH4)2SO4 precipitation. A hundred million leukocytes yielded 1.03 mg protein of the MPO preparation with the RZ of 0.31. The specific activity of the MPO preparation was about 29.25 u/mg. When 0.672 units of the MPO preparation were incubated with about 10(7) clinical isolates of Candida albicans in the presence of 0.2 mmol/L H2O2 and 0.14 mol/L NaCl. It was detected that 95.58 +/- 0.64% of the organisms were killed in the methylene blue staining system. PMID- 2551813 TI - Uterine herpes virus infection with multifocal necrotizing endometritis. AB - A case of herpes simplex virus (HSV) infection was diagnosed by biopsy of the cervix and endometrium in a 28-year-old woman with abnormal uterine bleeding. The cervical biopsy demonstrated surface ulceration and underlying patchy necrosis of endocervical clefts and stroma. The endometrium was late secretory, with striking patchy necrosis of gland epithelium and stromal cells. Both sites contained occasional epithelial and stromal cells with nuclear inclusions consistent with HSV infection. Viral culture further confirmed the presence of HSV. Immunohistochemistry demonstrated the presence of HSV antigens in the tissue, and ultrastructural study of the endometrium revealed viral particles within epithelial and stromal cells. The results suggest endometrial involvement via an ascending infection from the cervix. Recognition of this unusual pattern of endometrial inflammation may facilitate diagnosis of additional cases. PMID- 2551814 TI - Infection-associated vascular lesions in acquired immunodeficiency syndrome patients. AB - Several reports have recently appeared in the literature describing "unique" non neoplastic vascular lesions in patients with the acquired immunodeficiency syndrome (AIDS). These lesions may be mistaken clinically and histologically for Kaposi's sarcoma. The terms epithelioid angiomatosis, epithelioid or histiocytoid hemangioma, and pyogenic granuloma have all been used to describe a similar entity in which cat scratch disease bacillus (CSDB) was subsequently identified. Lesions closely resembling this entity occur in patients with bartonellosis. We report a case of a cutaneous vascular lesion on the hand of an AIDS patient in which cytomegalovirus (CMV) and organisms consistent with CSDB were both found. Simultaneous infections with CMV and CSDB have not been previously described. The presence of these organisms in and around endothelial cells may provide the common stimulus for the formation of these reactive vascular proliferations. PMID- 2551815 TI - HPV DNA detection in cervical swabs. PMID- 2551816 TI - Determination of deletion sizes in the MHC-linked complement C4 and steroid 21 hydroxylase genes by pulsed-field gel electrophoresis. AB - In man, the genes encoding the complement component C4 (C4A, C4B) of the immune system and the steroid 21-hydroxylase enzyme (CYP21A, CYP21B) of adrenal steroid biosynthesis are located in the major histocompatibility complex (MHC). Frequent gene deletions and duplications have been described in the C4 and CYP21 genes, particularly in patients with autoimmune diseases and congenital adrenal hyperplasia. Here we report the determination of deletion sizes in 11 chromosomes with six different deletions. The deletions spanned the C4A+CYP21A, C4B+CYP21A, and C4B+CYP21B gene pairs as determined by standard Southern blot analysis. The deletion size fell within the range of 30-38 kb in all the chromosomes, as determined by pulsed-field gel electrophoresis. Because the deletion sizes in most other gene clusters are more heterogeneous, the results suggest the involvement of a specific mechanism in the generation of C4+CYP21 deletions. PMID- 2551817 TI - Interaction of different strains of Entamoeba histolytica with target cells: characterization of electrophysiological and morphological features. AB - Two strains of Entamoeba histolytica with pathogenic zymodemes (SFL3, HK9), one strain with non-pathogenic zymodeme ("Bru") and one non-pathogenic Entamoeba sp. strain ("cold strain"), were investigated with respect to their interaction with target cells. Three test systems were used: 1) direct microscopical observation and qualitative as well as quantitative evaluation of contact and binding events with MDCK cells as targets, 2) kinetics of cytotoxic activity as measured by means of chromium release from 51Cr-labelled K562 cells, and 3) electrophysiological observations with freshly prepared mouse liver cells. We observed that the non-pathogenic cold strain interacted only shortly with target cells (statistical events, interaction type "I"), but did not induce morphological changes, chromium release or depolarization of targets. Non pathogenic and avirulent strain "Bru" showed, apart from type "I"-binding, the ability to establish tight (type "II") and long-lasting contact (type "III") with targets, but again without cytotoxic effects. The pathogenic but avirulent strain HK9 tightly interacted (type "II") and sometimes long-lasting with target cells, but morphological changes and chromium release were of a moderate degree during the first 20 min, and depolarization was only a rare event. In contrast, strain SFL3 produced tight and long-lasting contacts (type "III" binding), leading to cell death in 83% (type "IV" interaction) within 20 min, substantial chromium release within 10 min and rapid depolarization ("electric collapse") of target cells. PMID- 2551818 TI - Adjuvant activity of respirable iron ore dust. AB - The influence of the respirable fraction of an iron ore dust on immunity has been studied. The iron ore dust enhanced the specific immune responses to both sheep erythrocytes and a bacterial lipopolysaccharide. The IgG class of antibody was enhanced significantly and its pattern of enhancement suggested that the iron ore dust was functioning as an adjuvant. In studies to test this possibility further, the pattern of antibody development over both the primary and the secondary immune response to sheep erythrocytes closely matched that of Freund's incomplete adjuvant and low levels of silica. The splenocytes, from animals that had received iron ore dust for various lengths of time, exhibited enhanced polyclonal mitogenic activity for phytohaemagglutinin, in vitro. These properties were seen when the dust had been inhaled or injected into the pleural cavity and persisted for over 1 year following the injection or inhalation of the dust. Attempts to show the induction of interleukin-1 by the macrophages from mice implanted intraperitoneally with the iron ore dust were unsuccessful. PMID- 2551819 TI - Susceptibility to a mouse acquired immunodeficiency syndrome is influenced by the H-2. AB - The development of a mouse acquired immunodeficiency syndrome (MAIDS) induced following LP-BM5 MuLV infection depends on host genetic factors. Susceptible mice, such as C57BL/6J mice, develop a profound impairment of lymphoproliferative response to mitogens and hyperplasia of lymphoid organs and succumb to infection within 6 months. These changes do not occur in resistant mice, such as A/J mice. Resistance to MAIDS is a dominant trait since (C57BL/6J x A/J)F1 hybrid mice did not develop any immune dysfunctions following infection. Genetic regulation of the trait of resistance/susceptibility to MAIDS was determined in AXB/BXA recombinant inbred (RI) mouse strains (derived from resistant A/J and susceptible C57BL/6J progenitors). Two different criteria were used to determine their resistance or susceptibility to developing MAIDS: the gross pathologic evaluation of lymphoid organs at 13-15 weeks of infection, and survival. RI mouse strains segregated into two non-overlapping groups. The first group did not develop any significant pathology, and these mouse strains were considered as resistant to MAIDS. The second group showed the virus-induced pathological changes as well as an immunological dysfunction as seen in C57BL/6J progenitor mice, and these strains were thus considered as susceptible to MAIDS. This bimodal strain distribution pattern of resistance/susceptibility to MAIDS among the RI strains suggests that this phenotype is controlled by a single gene. Linkage analysis with other allelic markers showed a strong association between resistance/susceptibility to MAIDS and the H-2 complex. Possession of the H-2b haplotype derived from C57BL/6J mice was associated with susceptibility to MAIDS, while the H-2a haplotype conferred resistance to the disease. This finding was confirmed by demonstrating that H-2a congenics on the susceptible C57BL/10 background were as resistant to MAIDS as A/J mice which donated the H-2a locus. Gene(s) within the H-2 complex thus represent the major regulatory mechanism of resistance/susceptibility to MAIDS. PMID- 2551820 TI - Effect of converting enzyme inhibition on renin synthesis and secretion in mice. AB - We have investigated the relative importance of renal renin stores and de novo synthesis during stimulation of renin secretion and the role of transcription and posttranscriptional factors in providing increased synthesis of renin. When enalapril was administered to previously untreated mice, plasma renin concentration increased 40-fold within 1.5 hours, and remained at a high level for the 8 days of the experiment. Renal renin decreased by 82% after 24 hours and thereafter increased to levels higher than controls. Calculations of renin turnover, based on data for the rate of metabolism of renin in plasma, indicated that most of the renin released in the first 24 hours could be accounted for by the decrease in renal renin stores, indicating that de novo synthesis played only a minor role. After 24 hours, however, when both plasma renin concentration and renal renin increased, the calculated rate of renin synthesis increased to nearly 40 times the rate in controls. When enalapril was administered to mice that had been depleted of plasma and renal renin by chronic sodium loading, plasma renin concentration increased markedly within 1.5 hours, but to only half the level achieved in the previously untreated mice. No decrease in renal renin occurred, suggesting that the renal renin remaining after chronic sodium loading was not available for release. Renal renin messenger RNA increased 4.5-fold after 6 hours, and after 8 days had increased to 5.0 times the level at day 0. The increase in calculated rate of renin synthesis was maximal between 5 and 8 days, when it was 54 times greater than at day 0. During enalapril treatment, there were marked increases in the granulation of the juxtaglomerular cells and in the amount of rough endoplasmic reticulum and Golgi apparatus they contained. These results suggest that posttranscriptional factors play a major role in determining the rate of renin synthesis. PMID- 2551821 TI - Angiotensin-induced hypertension in the rat. Sympathetic nerve activity and prostaglandins. AB - To elucidate mechanisms of angiotensin II (Ang II)-related hypertension, we infused angiotensin (76 ng/min s.c.) into rats with minipumps for 10-14 days. Control rats received sham pumps. We measured blood pressure by tail-cuff, and the excretion of aldosterone and prostaglandins (PG) (PGE2, prostacyclin derivative 6kPGF1 alpha, and thromboxane [Tx] derivative TxB2). Angiotensin II increased blood pressure by 20 mm Hg by day 2 and by 90 mm Hg by day 10. Aldosterone excretion increased from 10 to 70 ng/day in Ang II rats by day 7. Urine PGE2 did not increase in angiotensin rats; however, both 6kPGF1 alpha and TxB2 excretion increased with angiotensin. Control rats had no changes in any of these parameters. A sympathetic component was tested in a separate group of angiotensin rats that received phenoxybenzamine (300 micrograms/kg/day) during angiotensin infusion; their increase in blood pressure of 40 mm Hg at 10 days was less than in those rats with angiotensin alone but more than in control rats. Phenoxybenzamine did not influence the angiotensin-induced increases in excretion of 6kPGF1 alpha or TxB2. Additional groups of conscious angiotensin and control rats were equipped with splanchnic nerve electrodes on day 14 for recording of sympathetic nerve activity. Angiotensin rats had greater basal sympathetic nerve activity than the control rats. Incremental methoxamine injections demonstrated altered baroreceptor reflex function in rats receiving angiotensin. We conclude that increased blood pressure with chronic angiotensin infusion is accompanied by increased production of aldosterone and increased sympathetic tone. The latter may be modulated by PG. PMID- 2551822 TI - Atrial natriuretic factor in NaCl-sensitive and NaCl-resistant spontaneously hypertensive rats. AB - Our previous studies demonstrated that chronic dietary NaCl supplementation is associated with significant increases in plasma atrial natriuretic factor in Wistar-Kyoto (WKY) rats but not in NaCl-sensitive spontaneously hypertensive rats (SHR-S). The current study tested the hypotheses that 1) acute volume-induced atrial natriuretic factor release is impaired in SHR-S compared with control NaCl resistant SHR (SHR-R) and WKY rats maintained on basal (1%) NaCl diets; 2) dietary NaCl supplementation (8% NaCl for 2 weeks) alters acute volume-dependent atrial natriuretic factor release in these strains; and 3) replacement of the deficiency in circulating atrial natriuretic factor seen in NaCl-supplemented SHR S can reverse the NaCl-sensitive component of hypertension. SHR-S and control SHR R and WKY rats were placed on 1% or 8% NaCl diets at age 7 weeks; 2 weeks later, right atrial pressure and plasma atrial natriuretic factor were measured in conscious rats before and after acute volume expansion (7, 20, and 60 ml/kg, 5% dextrose, for 1 minute). The slopes of the right atrial pressure x plasma atrial natriuretic factor linear regression for the SHR-S fed both 1% and 8% NaCl were significantly shallower (p less than 0.01) than those of 1% NaCl-fed SHR-R or WKY rats. Dietary NaCl supplementation did not alter right atrial pressure in any strain and blunted acute volume-induced atrial natriuretic factor release in WKY rats, but not in SHR-S or SHR-R, suggesting the dietary NaCl-induced elevation in plasma atrial natriuretic factor levels in WKY rats may be related to impaired clearance, as well as enhanced release, of the peptide. The plasma levels of exogenous atrial natriuretic factor required to abolish the NaCl-induced pressor effect in SHR-S were 12-fold greater than endogenous plasma atrial natriuretic factor levels in 8% NaCl-fed WKY rats, suggesting that impairment of atrial natriuretic factor release does not play a major role in the pathogenesis of NaCl sensitive hypertension in SHR-S. PMID- 2551823 TI - Calcium channel alterations in genetic hypertension. AB - We proposed earlier that voltage-dependent calcium (Ca2+) current is altered in single azygos venous cells from Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR). In this study, the effects of different intracellular concentrations of ethylene glycol-bis-N,N,N',N',-tetraacetic acid (EGTA) on Ca2+ currents were investigated. Vascular muscle cells from SHR and WKY rats were equilibrated with pipette solution containing 0.1 mM or 10 mM EGTA. Increasing the EGTA concentration from 0.1 to 10 mM in SHR vascular cells significantly enhanced the peak amplitude of the longer lasting (L) current from 87 +/- 12 pA to 152 +/- 8 pA, while the transient (T) current amplitude was not significantly different (52 +/- 7 pA and 36 +/- 7 pA, respectively). In WKY rat vascular muscle cells, the amplitudes of the T and L currents were not significantly different with the same comparison of intracellular EGTA concentrations. These observations suggest that relatively low intracellular Ca2+ concentrations can more strongly modulate Ca2+ current through the L channel in SHR than WKY rat vascular muscle cells. PMID- 2551824 TI - Tetrandrine and transmembrane signal transduction: effect on phosphoinositide metabolism, calcium flux and protein kinase C translocation in human lymphocytes. AB - Time-course and dose-dependent studies showed consistent suppression of phosphoinositide turnover in Con A-stimulated human lymphocytes in the presence of the plant alkaloid, tetrandrine. Significant inhibition of Con A-stimulated calcium flux was also observed. Furthermore, protein kinase C activity was also significantly inhibited by tetrandrine irrespective of whether Con A or phorbol myristate acetate was the stimulant. These results suggest that the immunosuppressive properties of tetrandrine are in part mediated by the capacity of tetrandrine to interfere with transmembrane signalling. PMID- 2551825 TI - Actions of the protease inhibitor phenylmethylsulfonyl fluoride on neutrophil granule enzyme secretion and superoxide production induced by fMet-Leu-Phe and phorbol 12-myristate-13-acetate. AB - The protease inhibitor, phenylmethylsulfonyl fluoride inhibits granule enzyme release and, above 1 mM, superoxide production from rabbit peritoneal neutrophils induced by the chemotactic peptide, fMet-Leu-Phe. At concentrations below 1 mM, it enhances superoxide production. Superoxide generation stimulated by phorbol 12 myristate-13-acetate is increased by phenylmethylsulfonyl fluoride at all concentrations studied. Phenylmethylsulfonyl fluoride has no effect on the rise in intracellular calcium or the depolarization induced by fMet-Leu-Phe but does decrease the extent of repolarization and abolishes hyperpolarization. It depresses actin polymerization and abolishes cytoplasmic alkalinization caused by fMet-Leu-Phe. The increased phosphorylation induced by phorbol 12-myristate-13 acetate in four of the five proteins studied was not affected by phenylmethylsulfonyl fluoride, but the increased phosphorylation of the fifth, a 21-kD protein was enhanced. We conclude that phenylmethylsulfonyl fluoride acts on inhibitory and enhancing processes or steps induced by fMet-Leu-Phe which are subsequent to or independent of calcium mobilization and protein kinase C activity. PMID- 2551826 TI - Neural cell adhesion molecule (NCAM) is the antigen recognized by monoclonal antibodies of similar specificity in small-cell lung carcinoma and neuroblastoma. AB - We describe reagents from 2 workshops which had been identified as recognizing the same or very similar antigens based on their tissue reactivity. Examination of their tissue specificity led us to the conclusion that this was similar to the expression of the neural cell adhesion molecule (NCAM). We also describe the use of a transfection-based assay to show that these reagents do recognize NCAM. 3T3 cells were transfected with a full-length clone of human NCAM. Indirect immunofluorescence studies showed binding of all related antibodies to the transfectants, but not to the control 3T3 cells. In addition, biochemical analysis using certain antibodies in the cluster confirm that they detect NCAM in the transfectants. Our study shows the benefits of using workshops to compare monoclonal antibodies and a molecular approach to define the antigens recognized by such reagents. PMID- 2551827 TI - In vitro infection of cell lines with HTLV-I and SIVmac results in altered intracellular free calcium concentration and increased membrane polarization. AB - This study was designed to determine whether retrovirus infection alters basal concentrations of intracellular free calcium, the magnitude of intracellular free calcium change upon receptor-mediated cell-stimulation, and cell membrane polarization. Two retrovirus-infected cell lines were utilized: HTLV-I-infected MT-2 cells and SIV-infected H-9 cells. Uninfected H-9 cells were used as controls. The results show that the retrovirally-infected cells had an increased basal concentration of intracellular free calcium when compared with the control cells and also a reduced magnitude of intracellular free calcium response to receptor-mediated cell stimulants. The retrovirally-infected cell membranes were also hyperpolarized when compared with the control cells. PMID- 2551829 TI - Effects of DBcAMP on exercise capacity in patients with and without chronic heart failure. AB - The acute effects of vasodilation on exercise capacity in cardiac patients with (group 1) and without (group 2) congestive heart failure were studied using dibutyryl cyclic AMP (DBcAMP). Exercise was performed on an upright bicycle ergometer using a graded protocol. DBcAMP increased cardiac output and decreased pulmonary capillary wedge pressure and systemic vascular resistance both at rest and during maximal exercise in these two groups. However, before and after DBcAMP neither exercise duration (371 +/- 52 seconds vs. 388 +/- 44, NS, in group 1, 645 +/- 148 vs. 635 +/- 143 seconds, NS, in group 2, respectively) nor maximal oxygen consumption (12.8 +/- 2.3 ml/kg/min vs. 13.1 +/- 1.6, NS, in group 1, 20.3 +/- 1.4 vs. 20.1 +/- 1.5, NS, in group 2, respectively) was improved. In both groups the arteriovenous oxygen differences were lower at rest and during exercise performed while on DBcAMP than in the control state. In group 2 patients excess vasoconstriction mediated by abnormally increased neurohormonal activities or edema were absent. Failure of the vasodilator to increase exercise capacity is probably due to nonspecific vasodilation and maldistribution of increased cardiac output, and not to tight vasoconstriction or narrowed arteriolar lumen. PMID- 2551828 TI - Pacemaker activity is modulated by tissue levels of cyclic adenosine 3',5' monophosphate in human atrial fibers. AB - We studied the role of tissue cyclic AMP levels in the chronotropic effects of theophylline on automatic human atrial fibers obtained from the hearts of 17 patients undergoing corrective open-heart surgery. Atrial fibers were perfused with Tyrode solution and transmembrane action potentials were recorded with a conventional microelectrode technique. In normal Tyrode solution, theophylline (0.1-1 mM) often decreased the late diastolic slope and the spontaneous rate. In the presence of 0.3-1 microM epinephrine, however, theophylline dose-dependently increased the diastolic slope, the rate of spontaneous discharges and the force of contraction. The increase in tissue level of cyclic AMP (+288 +/- 69%) induced by 0.3 mM theophylline in the presence of epinephrine was much greater than the increase (+73 +/- 19%) in the absence of epinephrine. It is concluded that pacemaker activity in human atrial fibers is modulated by tissue levels of cyclic AMP and theophylline may induce atrial tachycardia through an increase in the diastolic slope and the rate of discharges of automatic atrial fibers. PMID- 2551830 TI - Relapse and recovery in drug abuse: research and practice. AB - Relapse and recovery are two related areas which are central to drug abuse treatment research and practice. The view of drug addiction and dependence as a chronic, relapsing disease is supported by both research and clinical experience. Recent research shows that many drug abusers have relatively short "addiction careers" and suggests that a better understanding of relapse (and its prevention) may assist treatment providers in more effectively moving clients toward recovery. PMID- 2551831 TI - Site-directed mutagenesis at amino terminus of recombinant ricin A chain. AB - Successful immunotoxin therapy may depend upon reduction of the size of the components in order to decrease antigenicity and rate of clearance. In initial attempts to modify the A chain of ricin by deletion analyses within a prokaryotic expression system, coding sequences were modified by the insertion of unique restriction endonuclease sites and by the removal of 22 codons near the 5' terminus. The work presented here examines the expression, solubility, and activity of these mutant proteins and demonstrates that while amino acid residues may be altered in this region, the deletion of residues 19 through 40 yields an insoluble and inactive toxin molecule. PMID- 2551832 TI - Circular dichroism of bovine serum albumin in divalent salt solutions. AB - The circular dichroism of bovine serum albumin in divalent salt solutions was investigated. The salts selected were magnesium chloride and calcium chloride. Their effects on the secondary and tertiary structures of the protein were compared with that of lithium chloride. It is well known that the elements Ca, Mg, and Li have many properties in common. The results show the similarity of their ions in the capacity of deforming protein, in spite of their characteristic pharmacological functions. PMID- 2551833 TI - Opioid receptor selectivity of peptide models of beta-endorphin. AB - Two peptides, designed to contain structural models of the proposed hydrophilic linker domain (residues 6-12) and amphiphilic alpha-helical domain (residues 13 29) in beta-endorphin, have been tested for their abilities to mimic the opioid receptor selectivity profile of the natural hormone. In competitive binding assays employing guinea-pig brain membranes, both peptides displayed a much higher affinity for mu- and delta-opioid receptors than for kappa opioid receptors. Relative to beta-endorphin, the peptide models were 2-3 times more potent in the mu and kappa receptor binding assays, and about equipotent in the delta receptor binding assay. In guinea-pig ileum assays, one peptide was equipotent to beta-endorphin and the other was twice as potent. Like beta endorphin, their actions on this tissue were highly sensitive to naloxone antagonism, indicating that they were mediated by mu receptors and not kappa receptors. In view of the design of the two peptide models, and their minimal homology to the natural hormone, these results provide additional evidence in support to our proposal for the functional conformation of beta-endorphin. PMID- 2551834 TI - [A 50-year-old patient with precordial pressure sensation and increased dyspnea in a unilateral hyperlucent lung]. PMID- 2551835 TI - 45Ca uptake by retinal pigment epithelial cells. AB - Uptake of 45Ca was studied in isolated frog retinal pigment epithelial cells. 45Ca accumulation was found to be a saturable, temperature-dependent event. Kinetic analysis of this accumulation revealed two transport systems with apparent km of 2.0 and 0.3 mM. We found the presence of a Na-Ca exchanger mechanism that releases Ca2 under depolarized conditions. Light induced an increase of 45Ca uptake due to activation of the Na-K-ATPase and consequent decrease of extracellular potassium concentration. PMID- 2551837 TI - Initial characterization of lens beta-adrenergic receptors. AB - Crude plasma membranes were prepared from chick lens epithelial and fiber cells and assayed for specific binding of the beta-adrenergic antagonist dihydroalprenolol. Both membranes specifically bound the ligand, with the epithelial membranes exhibiting a greater number of higher affinity sites. This is the first demonstration of lens beta-adrenergic receptors using this type of assay and the first direct demonstration of beta-adrenergic receptors in lens fiber cells. PMID- 2551836 TI - Cyclic nucleotide modulation of herpes simplex virus latency and reactivation. AB - Clinical observations and experimental studies suggested that the relative proportions of ganglionic neuronal intracellular cyclic adenosine monophosphate (c-AMP) and cyclic guanosine monophosphate (c-GMP) concentrations may influence the state or activity of herpes simplex viral DNA in its relationship with the host cell DNA. We studied the effects of putative modulators of intracellular cyclic nucleotide levels on herpes simplex virus (HSV) reactivation from latency in murine trigeminal ganglion cells. We also investigated the effects of these same mediators on the c-GMP and/or c-AMP concentrations in HSV-latently infected trigeminal ganglion cells and in acyclovir-suppressed, HSV-infected neuroblastoma cells. Cholera toxin and theophylline increased c-AMP levels (2-fold and 5-fold at 1 min and 30 sec, respectively for cholera toxin and 2-fold and 1.5-fold at 1 min and 30 sec for theophylline) and enhanced the rapidity of HSV reactivation from latency (P less than 0.005). Exogenous dibutyryl c-AMP also stimulated viral reactivation (P less than 0.005). Carbamylcholine increased c-GMP levels (7-fold and 6-fold at 15 sec and 30 sec, respectively), produced no significant change in c-AMP levels, and delayed HSV reactivation from latency (P less than 0.005). None of these mediators had a demonstrable effect on HSV replication. PMID- 2551838 TI - Localization of angiotensin converting enzyme in the ciliary epithelium of the rat eye. AB - Angiotensin converting enzyme (ACE, E.C. 3.14.5.1) was localized in the rat eye by immunocytochemical staining using anti-rat lung ACE monoclonal antibody, and by autoradiography using the potent ACE inhibitor [3H]captopril. Highest levels of ACE immunoreactivity and [3H]captopril binding were found in the ciliary epithelium (Bmax = 1050 fmol [3H]captopril bound/mg protein) while lower levels were present in the sclera (Bmax = 500 fmol/mg protein). Scattered regions of the choroidal epithelium were weakly immunocytochemically stained by the anti-ACE antibody. No other major sites of labeling of ACE were demonstrated. In the ciliary epithelium, ACE was confined to cells bordering on the posterior chamber of the eye, similar to its presence at the ventricular surface of the choroid plexus of the brain. These findings suggest that ACE may metabolize a peptide involved in the regulation of aqueous humor production. PMID- 2551839 TI - Activation of beta-adrenergic receptors causes stimulation of cyclic AMP, inhibition of inositol trisphosphate, and relaxation of bovine iris sphincter smooth muscle. Biochemical and functional interactions between the cyclic AMP and calcium signalling systems. AB - We have investigated the interactions between the cAMP and inositol 1,4,5 trisphosphate (IP3)-Ca2+ signalling systems in the bovine iris sphincter by measuring the effects of beta-adrenergic and cholinergic muscarinic agonists and antagonists on cAMP formation, IP3 accumulation and muscle contraction relaxation. (1) Addition of 5 microM isoproterenol (ISO) or forskolin (5 microM) consistently produced stimulation of cAMP (540%), inhibition of IP3 (34%) and complete relaxation of the muscle. The ISO effects were dose-dependent, with EC50 values for cAMP formation, IP3 inhibition and muscle relaxation of 2.8 X 10(-7) M, 3.4 X 10(-7) M and 0.45 X 10(-7) M, respectively. (2) Timolol, a beta adrenergic antagonist, inhibited the ISO effects in a dose-dependent manner, with IC50 values for cAMP formation, IP3 accumulation and muscle relaxation of 1.8 X 10(-5) M, 3.2 X 10(-5) M and 2.2 X 10(-5) M, respectively. (3) The effects of ISO (0.5 microM) were time-dependent, and they clearly indicate a temporal relationship between the agonist-induced stimulation of cAMP, inhibition of IP3, and relaxation of the muscle. Within 15 sec following the addition of ISO, there was a marked increase in the level of cAMP, a decrease in IP3, and this was accompanied by an equally rapid relaxation of the muscle. (4) Addition of carbachol (CCh) to iris sphincter pretreated with ISO decreased cAMP formation and reversed muscle relaxation to complete contraction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551840 TI - Glucose-6-phosphatase activity in the retina of the awake rat. AB - In the 2-deoxyglucose technique, the rate of glucose utilization in small areas of the central nervous system is measured using a standard operational equation that assumes negligible glucose-6-phosphatase activity. Hoping to apply this technique in the mammalian retina, we sought to identify the extent of this enzyme's activity in vivo. [2-3H] glucose but not [U-14C] glucose loses its label during metabolism and returns to the glucose precursor pool in the presence of glucose-6-phosphatase. Accordingly, a decline of 3H/14C in the retinal glucose pool with time indicates glucose-6-phosphatase activity. We injected a mixture of [2-3H] glucose and [U-14C] glucose into the internal carotid artery of 10 awake rats via a previously inserted catheter. Plasma samples were collected and the eyes enucleated at timed intervals. The eyes were immediately frozen, freeze dried and dissected to obtain retina. Radiolabeled glucose was separated using ion exchange and paper chromatography prior to scintillation counting. The 3H/14C ratio was found to decline at a statistically significant rate of about 2.5% per minute, indicating glucose-6-phosphatase activity. However, an estimate of the turnover of retinal glucose suggests that glucose-6-phosphatase dephosphorylates a minimal percentage of the glucose entering the glycolytic pathway, allowing application of the standard operational equation to the mammalian retina in vivo. PMID- 2551841 TI - Phase II trial of spirogermanium in advanced non-small cell lung cancer. An Illinois Cancer Council Study. AB - A phase II trial of spirogermanium was conducted in advanced previously untreated non-small cell lung cancer patients. The drug was given by intravenous infusion 3 times per week for 2 weeks, twice per week for the next 2 weeks, and then weekly. Starting dose was 125 mg/m2, and dose escalation of 25 mg/m2 per week was required in the absence of toxicity to a maximum dose of 200 mg/m2 per infusion. Fifteen eligible patients were treated, and no objective responses were seen. Primary toxicity was neurologic and reversible after withdrawal of the drug. We conclude that spirogermanium is not active against non-small cell lung cancer in the dosage used in this study. PMID- 2551843 TI - Deeper 'Go' phase in rat fibroblasts affects the lag time required for T antigen induced DNA synthesis, but not the lag time required for SV40 T antigen expression. AB - In density-arrested rat 3YltsG125 cells, the time required for entry into S phase after serum stimulation is prolonged with increase in duration of the arrest. When these cells were infected with simian virus 40, the time required for expression of T antigen did not change, but the time required for entry into S phase was prolonged with increase in duration of the arrest. The extent of prolongation of the interval between T antigen expression and entry into S phase correlated well with the extent of the prolongation detected with serum stimulation. These observations suggest that: (1) expression of T antigen was not affected by the level of cellular preparedness for entry into S phase, i.e., it did not depend on cellular functions directly involved in the preparedness, and (2) T antigen induced cellular DNA synthesis, at least in part, by a mechanism similar to that of serum-induced DNA synthesis. PMID- 2551842 TI - Evidence for attachment of fatty acid to varicella-zoster virus glycoproteins and effect of cerulenin on the maturation of varicella-zoster virus glycoproteins. AB - Varicella-zoster virus (VZV) contains four major glycoproteins designated gpI, gpII, gpIII, and gpIV. In the present study the presence and role of fatty acids in these glycoproteins were examined. The incorporation of fatty acid into the glycoproteins that span the membranes of VZV was studied. For this, virus infected human embryo fibroblast cells were labeled with [3H]-palmitic acid, an extract of the cells was treated with monoclonal antibodies to VZV glycoproteins, and the precipitate was analyzed by SDS-PAGE. The electrophoretic pattern showed that gpI was heavily labeled and that gpII and gpIV were lightly labeled. Cerulenin, an antibiotic that inhibits de novo fatty acid biosynthesis, was used to examine the effects of fatty acids on replication of VZV and posttranslation processing of glycoproteins. Treatment of the cells with cerulenin significantly inhibited viral growth, but did not affect protein synthesis in the cells. Examination of processing of viral glycoproteins in these cerulenin-treated cells revealed the accumulation of precursor proteins and a decrease in the amount of mature glycoproteins. These data suggest that fatty acid acylation of VZV glycoproteins is necessary for formation of complete infectious virions. PMID- 2551844 TI - Radiochemical synthesis of [18F]fluororaclopride. AB - The radiochemical synthesis of [18F]Fluororaclopride (S-3,5-dichloro-6-methoxy-N (1-(2-[18F]fluoroethyl)-2-pyrrolidinylmet hyl) salicylamide) is accomplished via a two step synthesis. [18F]Fluoroethyltriflate is prepared by [18F]fluoride displacement on the bis triflate of ethylene glycol. [18F]Fluoroethyl triflate is then allowed to alkylate a secondary amine precursor to yield [18F]fluororaclopride. Purification of the radiopharmaceutical involves use of a short silica BONDELUT column and subsequent reverse-phase HPLC. The final product is obtained with a radiochemical yield of approximately 15% (corrected for decay) in a synthesis time of approximately 50 min. Fluororaclopride displays a 3- to 4 fold lower affinity for the D2 receptor than does raclopride. PMID- 2551845 TI - Alpha activity measurement of cigarettes in Kuwait. AB - A semiconductor surface barrier detector with an active area of 450 mm2 and depletion layer thickness of 300 microns has been used to measure the alpha activity of cigarette tobacco. The performance parameters of the counting system, the calibration and the results of alpha-activity in some cigarettes are presented. PMID- 2551846 TI - Synthesis of radioiodinated N-succinimidyl iodobenzoate: optimization for use in antibody labelling. AB - N-succinimidyl-3-(tri-n-butylstannyl)benzoate (m-BuATE), N-succinimidyl-3-(tri methylstannyl)benzoate (m-MeATE) and N-succinimidyl-4-(tri-n butylstannyl)benzoate (p-BuATE) were synthesized and radioiodinated using either N-chlorosuccinimide (NCS) or t-butylhydroperoxide (TBHP) as the oxidant. Radiohalogenation of m-MeATE proceeded more rapidly than m-BuATE. NCS was the more efficient oxidant at reaction times less than 15 min; use of both TBHP and NCS resulted in nearly quantitative yields after 15 min when m-MeATE was used. Using NCS, achieving optimal antibody coupling and specific binding required purification of the active ester by HPLC; in contrast, with TBHP, only Sep-Pak purification was needed. PMID- 2551847 TI - Paraffin scintillator for radioassay of solid support samples. AB - A new paraffin scintillator used for solid support sample counting has been proposed, and its composition and various characteristics are described. The solid support sample treated with this scintillator can be easily handled because of rigid sample conditions. This technique provides great advantages such as the elimination of a large volume of scintillator and little radioactive waste material by using an economical polyethylene bag instead of the conventional counting vial. PMID- 2551848 TI - Determination of the lysosomal role in tumor accumulation of 67Ga by dual-tracer studies. AB - The lysosomal role in tumor accumulation of 67Ga was determined by dual-tracer (67Ga and 46Sc) studies. It became clear that 67Ga essentially did not accumulate in the tumor lysosome, and that the lysosome did not play a major role in tumor accumulation of 67Ga. In addition, it was revealed that tumor lysosome was hardly disrupted at all in some phases of fractionation procedures. PMID- 2551849 TI - Indirect spectrophotometric determination of BIDA, DISIDA, DTPA and MDP in labelled compounds. AB - N-(4-(n-butyl)-acetanilide)iminodiacetic acid (BIDA), N-(2,6 diisopropylacetanilide)iminodiacetic acid (DISIDA), diethylenetriaminepentaacetic acid (DTPA) and methylene diphosphonic acid (MDP) are used in labelling kits. The contents of BIDA, DISIDA or MDP of the 99mTc-labelled compounds can be determined (indirectly) spectrophotometrically with copper, eriochrome cyaanine R (ECC) and dodecylethyldimethylammonium bromide (DEDA) in a sodium barbital buffered system at pH 8.5. The calibration curves obey Beer's Law from 0 to 40 micrograms/25 mL for BIDA and DISIDA, 0 to 60 micrograms/25 mL for DTPA and 0 to 100 micrograms/10 mL for MDP. PMID- 2551851 TI - The synthesis of 4-S-cysteinyl-[U-14C]phenol, an experimental antimelanoma agent. AB - 4-S-Cysteinylphenol (4-CP) has been shown to exert selective toxicity to melanocytes, causing growth inhibition of experimental malignant melanoma. 4-S Cysteinyl-[U-14C]phenol (4-[U-14C]CP) has now been synthesized by reaction of [U 14C]phenol with excess L-cystine in HBr. After crystallization of unreacted cystine from the reaction mixture, 4-[U-14C]CP was recovered by HPLC in 57.6% radiochemical yield with a specific activity of 26.2 MBq mg-1 (15.2 mCi mmol-1). Radiochemical purity was greater than 99%. 2-S-Cysteinyl-[U-14C]phenol (2-[U 14C]CP), a side product with no antimelanoma activity, was recovered in 6% radiochemical yield. PMID- 2551850 TI - Physico-chemical study of the radiopharmaceuticals 99mTc-DMSA, 99mTc-EDTA and 99mTc-DTPA interaction with plasmatic proteins. AB - This report studies the binding rate of the radiopharmaceuticals 99mTc-DTPA, 99mTc-EDTA and 99mTc-DMSA to plasmatic proteins. The proteins bind to the tested radiopharmaceuticals in the following sequence: 99mTc-DTPA less than 99mTc DMSA(C1) less than 99mTc-EDTA less than 99mTc-DMSA(C2) where C1 and C2 represent two different Tc-DMSA complexes. The thermodynamic study suggests a quantitative relationship of radiopharmaceutical:protein = 1:1 and an almost nonexistent influence of the temperature, which means that the interacting forces in this process are relatively weak. PMID- 2551852 TI - Rapid production and trapping of [18F]fluorotrimethylsilane, and its use in nucleophilic fluorine-18 labeling without an aqueous evaporation step. AB - A flow-through system is described for rapid purification of [18F]fluoride for nucleophilic substitution reactions. It depends on generation of [18F]fluorotrimethylsilane from aqueous 18F solutions and subsequent collection of the gas and its hydrolysis by base in near anhydrous acetonitrile. Potassium t butoxide plus Kryptofix 2.2.2 was found to be a more effective base than K2CO3 plus Kryptofix or than tetraethylammonium hydroxide. The procedure takes 2-3 min and is thus faster than controlled removal of water from [18F]fluoride by evaporation. The trapped and hydrolyzed 18F was used in several nucleophilic substitution reactions. The flow-through system is also convenient for recovery of fluoride from impure and otherwise unreactive solutions, including residues from large scale 18F syntheses, and can thus lead to more efficient use of 18F where production of the radionuclide is limited. PMID- 2551853 TI - A modified distillation technique for the recovery of tritium from blood. PMID- 2551854 TI - Technetium-99m spiperone dithiocarbamate: a potential radiopharmaceutical for dopamine receptor imaging with SPECT. AB - Spiperone dithiocarbamate (SPDC) was prepared by reacting spiperone with carbon disulfide followed by sodium hydroxide. SPDC was labelled with 99mTc by reduction of pertechnetate with formamidine sulfinic acid or sodium dithionite at alkaline pH, resulting in approximately 40% incorporation of 99mTc. The lipophilic complex was conveniently isolated at high specific activity and high radiochemical purity by extraction into dichloromethane, which was then evaporated and the residue was redissolved in a 1:3 mixture of ethanol and saline containing 0.1 mg/mL gentisic acid. Biodistribution studies following i.p. injection in rats showed low uptake of radioactivity in the brain, but striatum/cortex and striatum/cerebellum ratios were reduced by pretreatment with haloperidol. This agent may allow imaging of dopamine D-2 receptors using single-photon emission computed tomography (SPECT). PMID- 2551855 TI - 11C-labelling of dimethylphenethylamine in two different positions and biodistribution studies. AB - Dimethylphenethylamine (DMPA), a substrate for the B-form of the monoamine oxidase enzyme (MAO, EC 1.4.3.4), was labelled with 11C in two different positions; in the methyl group (M-DMPA) and in the phenethyl group (P-DMPA). M DMPA was prepared by N-alkylation of methylphenethylamine with [11C]methyl iodide and P-DMPA was prepared by N-alkylation of dimethylamine with [1-11C]phenethyl iodide. The radiochemical yields were 30-35% (M-DMPA) and 10% (P-DMPA), based on [11C]carbon dioxide, with overall synthesis times of 30-35 min (M-DMPA) and 50 min (P-DMPA). The compounds were isolated by semi-preparative HPLC and the radiochemical purity was in both cases greater than 99%. The biodistributions of 11C-labelled M-DMPA [correction of M-DPMA] and P-DMPA were studied in rat brain by dissection and in the brain of a Rhesus monkey by positron emission tomography (PET). PMID- 2551856 TI - Chemical form of tumor-tropic 99mTc-DL-homocysteine. AB - Analyses of the chemical forms of 99mTc-complexes provide important information for the development of a new tumor-tropic 99mTc-labeled radiopharmaceutical. We attempted to determine the chemical form of 99mTc-DL-homocysteine (99mTc-Hcy) which was previously reported to be tumor-tropic. By analyzing the functional residues of Hcy in the 99mTc-Hcy molecule, it was estimated that the sulfhydryl and amino residues participated in the chelate formation. Gel filtration analysis of 99mTc-Hcy indicated that its molecular size was bigger than that of 99mTc penicillamine monomer. The analysis also indicated that 99mTc-Hcy complex seemed to be a relatively small oligomer. Although an uncertainty remains on the valency of Tc in 99mTc-Hcy molecule and the accurate molecular size of this complex, its putative chemical form is described. PMID- 2551857 TI - Solid scintillation counting: a new technique for measuring radiolabeled compounds. AB - This report describes the theory and practice of anew solid scintillator technique for measurement of radiolabeled compounds useful in bioresearch. Solid scintillation counting is expected to replace liquid scintillation counting in certain applications involving non-volatile radiolabeled substrates. PMID- 2551858 TI - Advancements of radiation induced degradation of pollutants in drinking and waste water. AB - The radiation induced decomposition of some water pollutants such as chlorinated methanes, ethanes and olefines has been studied under various conditions. In addition to product analysis, spectroscopic and kinetic characteristics of CH3, C2H5 and their peroxides were also reinvestigated by pulse radiolysis techniques. Based on the experimental results, probable reaction mechanisms for the decomposition of some pollutant systems are presented. At the same time the present state of the research in this field is given. PMID- 2551859 TI - Soil sampling and 137Cs analysis of the Chernobyl fallout in Greece. AB - A total of 1242 samples of soil, collected over Greece, during the period May November 1986, were counted and analysed for 137Cs from Chernobyl fallout. The counting was performed using a NaI detector on-line to a microcomputer, moreover, 252 of the samples were also analysed using Ge detectors, for inter-comparison and also for the assessment of other long-lived isotopes in the fallout. The results show that 137Cs fallout from Chernobyl presents a remarkable geographical variability. The evaluated ground activity due to 137Cs deposition ranges between 0.01 and 137 kBq/m2. PMID- 2551860 TI - The binding of 99mTc(Sn)-MDP complexes to human serum albumin and other blood proteins determined with gel chromatography and ultrafiltration. AB - The binding of 99mTc(Sn)-MDP to human serum albumin and other blood proteins was investigated by gel chromatography and ultrafiltration. During gel chromatography dissociation of the 99mTc(Sn)-MDP-protein complex occurs: thus, it is not a suitable technique for the determination of protein binding. The values found with ultrafiltration have to be corrected for non-ultrafiltrable TcO2.nH2O. From the corrected values it can be concluded that binding of 99mTc(Sn)-MDP to blood proteins does not play a role in the biodistribution. PMID- 2551861 TI - Eaton-lambert syndrome associated with pituitary apoplexy--a case report. AB - A case of Eaton-Lambert syndrome associated with pituitary apoplexy is presented. It is thought to be the first reported case. The clinical features, electrophysiological findings and effects and side effects of therapeutic drugs are noted. Endocrinological dysfunctions are discussed in relation to pituitary disorder. PMID- 2551862 TI - Ontogeny of hormone-secreting cells of the rat pituitary gland: an immunocytochemical study on dissociated cells. AB - An immunocytochemical study was undertaken in foetal, prepubertal and mature rats to determine the time of differentiation of various types of adenohypophyseal cells during development. Freshly dissociated pituitary cells from foetal (18-21 days postconception), neonatal (from birth up to 30 days) and adult rats (more than 8 weeks) were characterized using immunocytochemical methods. All types of hormone-producing cells were present at day 18 postconception, although only 20% of the cells were immunolabelled. Adrenocorticotropin (ACTH)-secreting cells accounted for the highest number of hormone-positive cells. Growth hormone secreting cells increased remarkably from day 18 postconception onwards. Prolactin-secreting cells were not seen in the foetal adenohypophysis and did not start to increase until 10 days after birth, whereas by that time the number of ACTH, thyrotropin, follicle-stimulating and luteinizing hormone-secreting cells had stopped increasing. By day 30 after birth, 80-95% of the cells were immunoreactive. PMID- 2551863 TI - False-positive reactions associated with anti-mouse activity in serotests for feline leukemia virus antigen. AB - False-positive reactions were observed in commercial test kits designed to detect FeLV infections in cats. The false-positive reactions were associated with heterophilic antibody in cat serum directed primarily against mouse immunoglobulins. Of 2,830 sera tested, 579 were considered FeLV positive or equivocal via ELISA; of these, 10 were found that did not have FeLV antigen but did have anti-mouse antibody. With 95% confidence, 0.14% to 0.57% of cats would have such false-positive reactions in ELISA, because one in 283 (0.35%) cats had anti-mouse antibodies. PMID- 2551865 TI - Nonangiogenic and nonlymphomatous sarcomas of the canine spleen: 57 cases (1975 1987). AB - The case records of and histopathologic findings in 57 dogs with nonangiogenic and nonlymphomatous splenic sarcomas were reviewed. Splenic neoplasms in these dogs included leiomyosarcoma, fibrosarcoma, undifferentiated sarcoma, liposarcoma, osteosarcoma, chondrosarcoma, myxosarcoma, rhabdomyosarcoma, and fibrous histiocytoma. The clinical signs associated with splenic sarcoma included anorexia or decreased appetite, abdominal distention, polydipsia, lethargy, vomiting, weight loss, and weakness. An abdominal mass was detected in 86% of the dogs by use of abdominal palpation (63%), and/or abdominal radiography (74%). The diagnosis was based on histopathologic findings in the spleen. Abdominal exploratory surgery was performed on 43 of the 57 dogs. Twenty-seven dogs were treated by splenectomy, and 16 were euthanatized at the time of surgery because of widespread metastatic lesions. Of the 14 dogs on which surgery was not performed, 11 were euthanatized on the basis of results of preoperative diagnostic tests, and the remaining 3 dogs had splenic neoplasms that were incidental findings at necropsy. Of the 27 surgically treated dogs, 5 died in the immediate postoperative period, 12 died or were euthanatized within 1 year after splenectomy, and only 5 dogs survived greater than or equal to 1 year. Three dogs were lost to follow-up evaluation, and 2 were still alive 6 and 7 months after surgery. The median survival time of the 22 dogs for which survival was known was 2.5 months. The median survival time for 11 dogs with no obvious metastasis at the time of splenectomy was 9 months.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551864 TI - Sensitivity and specificity of blood test kits for feline leukemia virus antigen. AB - Enzyme-linked immunosorbent assay (ELISA) test kits have been used widely to diagnose FeLV infection. Several companies have licensed such kits, but because these test kits are designed somewhat differently, a demonstration of their relative merits was needed. Differences in the sensitivity and specificity of 7 commercial test kits were determined by testing sera that induce false-negative (limiting dilution of FeLV group specific antigen, p27), and false-positive (cat anti-mouse antibody) test results. Among the panels of sera used in this study, significant differences in test kit sensitivity were not observed, but differences in specificity were identified. An interim report of these studies, supplied to the manufacturers, prompted changes in some of the kits, resulting in improved specificities. Generally, FeLV test kits are now more efficient in accurately detecting p27 in blood, serum, or plasma of cats. Nevertheless, some kits still lack specificity attributable to reagents supplied by the manufacturer. PMID- 2551866 TI - Interpreting feline leukemia test results. AB - Routine prevaccinal screening for FeLV has inherent statistical limitations owing to the magnification of false-positive test errors by the low prevalence of FeLV viremia in the general cat population. Positive ELISA test results obtained in a screening program should be interpreted with caution, because a high proportion- approximately 72%--of such are likely to be false-positive results. On the other hand, routine screening is an excellent method for ruling out FeLV viremia, because a false-negative result is likely to be obtained in only 1/1,000 tests. PMID- 2551867 TI - Systemic autoimmune disease and concurrent nematode infection in a dog. AB - Systemic lupus erythematosus was diagnosed in a dog with concurrent nematode infection. The clinical signs of disease were unusually severe and included multiple neurologic deficits, polyarthritis, and weight loss. The dog was thrombocytopenic, and serotest results included positive lupus erythematosus test, positive rheumatoid factor test, positive antinuclear antibody test, hypergammaglobulinemia, and high platelet-associated IgG concentration. After treatment of hookworm, whipworm, and heartworm infections concurrently with corticosteroid and empiric treatment, the dog's condition improved. However, 10 days later, cyclophosphamide administration was necessary for continued immunosuppression. The dog was euthanatized because of progressive deterioration and development of canine coronavirus diarrhea. Serotest data generated from the dog's serum obtained at the time of referral suggested that autoantibodies and circulating immune complexes may have included IgE isotypes. PMID- 2551868 TI - Rearranged Epstein-Barr virus genomes and clonal origin in nasopharyngeal carcinoma. AB - Epstein-Barr virus (EBV) is known to be associated with two malignant diseases, nasopharyngeal carcinoma (NPC) and endemic Burkitt's lymphoma. In this study, the genomes of EBV in biopsy specimens from 4 NPC patients in Japan were analyzed using Southern blot hybridization. The NPC tissues of all examined cases contained rearranged EBV genomes whose BamHI H fragments were larger than those of prototype EBV genomes. One of them had a BamHI fragment containing contiguous sequences of BamHI Y and H. A single-sized EBV DNA terminus was observed in these NPC tissues, implying the evolution of the carcinoma from a single EBV-infected cell. PMID- 2551870 TI - Voluntary feed intake, acid-base balance and partitioning of urinary nitrogen in lambs fed corn silage with added sodium bicarbonate or sodium sesquicarbonate. AB - An experiment with growing lambs was designed to test the hypothesis that alterations in blood acid-base status would influence intake of corn silage. Six wethers (29 kg) were fed a diet of corn silage (36% DM, 8% CP) supplemented with 1.25% urea and .2% sulfur. At feeding time, sodium bicarbonate (NaHCO3) and sodium sesquicarbonate (NaSC) were added to the silage at levels of 0, 2% or 4% of diet DM. The treatments were arranged as a 2 x 3 factorial, and the study was conducted as a 6 x 4 incomplete latin square with four 17-d periods. Voluntary intake of OM was not different (P greater than .05) between NaHCO3 (1,008 g/d) and NaSC (1,041 g/d). There was no significant interaction between type of buffer (NaHCO3 or NaSC) and level of buffer on any of the variables measured. The progressive increase in buffer load did not alter feed intake (P greater than .05), although there was a quadratic response (P less than .05) in urine pH and a linear increase (P less than .01) in blood HCO3- 2 h after feeding. There was no evidence that lambs fed corn silage experienced metabolic acid stress. Urinary excretion of ammonia and urea were indicative of changes, although not pronounced, in ammoniuria and ureapoiesis in response to bicarbonate loading. This study implies that corn silage imposes no "acid stress" on lambs and, consequently, that there is no nutritional benefit in adding buffers to corn silage for sheep. PMID- 2551871 TI - A review of the use of intrinsically 14C and rare earth-labeled neutral detergent fiber to estimate particle digestion and passage. AB - Methodology that allows simultaneous measurement of dynamic events affecting NDF digestion in passage from the rumen should improve our understanding of factors influencing intake and digestion. Ideally, particle flow is measured with a marker indelibly attached to or intrinsically part of the feed. If flow measurements are to reflect physiological conditions, marked and unmarked feed must be digested and passed with identical fractional rates. Application of 14C labeled plant fiber to the study of ruminal dynamics has been slow because of expense and difficulty in producing 14C-labeled plant material. Recently, alfalfa was intrinsically labeled with 14C under field conditions to produce plant material similar in composition to unlabeled material. Carbon-14 specific activity was similar in all particle sizes, and in indigestible and digestible NDF. Greater concentrations of ytterbium (Yb) were associated with smaller vs larger particles. Larger differences in turnover rates among animals than differences attributable to treatments force comparisons of markers to be made within animal or with in vitro systems. The uncertainty about how extrinsic markers respond under various environments resulting from interaction of feed properties and gut function, and the high error inherent in measuring dynamic systems, raise serious questions on the interpretability of results. Advantages of 14C-labeled NDF over other markers include simultaneous measurements of particle breakdown, digestion and passage rates as well as the potential to study microbial attachment, VFA, CO2 and CH4 production and rate of incorporation of labeled metabolites into tissues. PMID- 2551872 TI - In-vitro susceptibility of Campylobacter pylori to quinolones. PMID- 2551869 TI - A case of leiomyosarcoma associated with humoral hypercalcemia of malignancy: demonstration of biological and immunological activities of parathyroid hormone related protein in the tumor extract. AB - Hypercalcemia occurred in a patient with leiomyosarcoma when multiple lung metastases developed. Despite normal plasma parathyroid hormone (PTH) levels and low 1,25-dihydroxyvitamin D, this hypercalcemic patient had a marked hypercalciuria and phosphaturia associated with an increased excretion of nephrogenous cyclic AMP (NcAMP). Administration of cisplatin ameliorated both the hypercalcemia and hypercalciuria without any reduction in tumor size of NcAMP excretion. Terminally, acute pancreatitis occurred producing a profound hypocalcemia. In the extract of tumor tissue obtained post mortem, bioactivity stimulating the generation of cyclic AMP in osteogenic cells was demonstrated along with the immunoreactive PTH-related protein (PTH-rP). the first report of a solid non-epithelial malignancy producing PTH-rP and associated with humoral hypercalcemia of malignancy. The hypercalcemia in this case caused acute pancreatitis, which led to a profound hypocalcemia. PMID- 2551873 TI - Contrasting effects of inflammatory stimuli on neutrophil and monocyte adherence to endothelial cells. AB - Leukocyte adherence to endothelial cells (EC) is an important early event in inflammatory responses, which are often characterized by a predominance of either neutrophils (PMN) or monocytes. However, there is little information concerning the molecular events important in leukocyte adherence to EC. Intracellular activation of protein kinase C and the calcium-second messenger system leads to the stimulation of a number of important functions in PMN and monocytes. We compared the effects of members of these pathways on human PMN and monocyte adherence to cultured bovine aortic EC. We observed that phorbol myristate acetate, phorbol, 12,13-dibutyrate, L-alpha-1-oleoyl-2-acetoyl-sn-3-glycerol, and ionomycin each induced significant dose-dependent increases in PMN adherence to EC monolayers. In contrast, similar concentrations of each of these agents induced significant decreases in EC adherence of monocytes enriched by countercurrent centrifugal elutriation. Separate experiments determined that the differences in PMN and monocyte adherence to EC were not related to differences in oxidant production because 1) phorbol myristate acetate and L-alpha-1-oleoyl-2 acetoyl-sn-3-glycerol caused similar marked increases in both PMN and monocyte superoxide anion and hydrogen peroxide production and 2) ionomycin, which had opposing effects on PMN and monocyte adherence, had no effect on PMN and monocyte superoxide anion or hydrogen peroxide release. We conclude that activators of protein kinase C and the Ca-second messenger pathway have opposite effects on PMN and monocyte adherence to EC and that these effects are mediated by O2 radical independent mechanisms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551874 TI - Effects of angiotensin blockade on the splanchnic circulation in normotensive humans. AB - The effects of angiotensin-converting enzyme inhibition (ACE-I) by enalapril on splanchnic (n = 10) and central hemodynamics (n = 9) were examined in moderately salt-depleted healthy volunteers, at rest and during 15-20 min of lower body negative pressure (LBNP), reducing mean arterial pressure by 10 mmHg. During LBNP before ACE-I, both splanchnic and total peripheral vascular resistances increased. During ACE-I, splanchnic and total peripheral vascular resistances decreased. After enalapril administration, splanchnic vascular resistance did not increase during LBNP. Total peripheral vascular resistance still increased but not to the same extent as during LBNP before ACE-I. The increases in heart rate and plasma norepinephrine during LBNP were attenuated after ACE-I compared with LBNP before ACE-I. The effectiveness of the ACE-I was clearly demonstrated by unchanged and low plasma angiotensin II levels during ACE-I. We conclude that, in normal sodium-depleted humans, acute ACE-I decreases splanchnic vascular resistance at rest and abolishes splanchnic vasoconstriction during LBNP. Furthermore, it may interfere with autonomic nervous system control of the circulation. PMID- 2551875 TI - Essential fatty acid-deficient rats are resistant to oleic acid-induced pulmonary injury. AB - Because leukotrienes and prostaglandins are inflammatory mediators derived from arachidonic acid, their potential role in oleic acid-induced lung injury was evaluated in control and in essential fatty acid-deficient (EFAD) rats depleted of arachidonic acid substrate. In control rats, oleic acid (0.06 ml/kg iv) increased the pulmonary permeability index (measured by scintigraphy) from -10 +/ 13 x 10(-6) s-1 to 217 +/- 20 x 10(-6) s-1 and 118 +/- 13 x 10(-6) s-1 at 5 and 50 min (P less than 0.05), respectively. It also caused arterial hypoxemia at 30 min (P less than 0.05). Compared with saline controls, oleic acid increased bronchoalveolar lavage fluid levels of immunoreactive (i) LTC4/D4, iLTB4, (P less than 0.01), and 6-ketoprostaglandin F1 alpha (6-keto-PGF1 alpha) (P less than 0.05). In EFAD rats, oleic acid failed to significantly increase the lung permeability index at 5 and 50 min. In contrast to control rats, oleic acid failed to cause hypoxemia in the EFAD rats. Bronchoalveolar lavage levels of iLTB4 and i6-keto-PGF1 alpha after oleic acid in EFAD rats were lower compared with oleic acid controls, whereas iLTC4/D4 in the oleic acid EFAD group was not decreased. Treatment with intraperitoneal ethyl arachidonate (400 mg over 2 wk) reversed the resistance of EFAD rats such that the pulmonary edema (P less than 0.05) was evident after oleic acid. This latter group also manifested a significant (P less than 0.05) rise in the bronchoalveolar lavage levels of iLTB4 and i6-keto-PGF1 alpha. These results suggest that arachidonic acid metabolites contribute to oleic acid-induced pulmonary permeability. PMID- 2551876 TI - Effect of dimethylthiourea on the neutrophil myeloperoxidase pathway. AB - The sulfur-centered compound dimethylthiourea (DMTU) affords antioxidant protection in animal models of acute lung injury, an effect that has been attributed to its OH. scavenging properties. Although DMTU can also react with H2O2 in certain experimental systems, the effect of DMTU on the neutrophil myeloperoxidase (MPO) pathway has not been studied. DMTU (1-10 mM) completely blocked stable oxidants and hypochlorous acid formation by phorbol myristate acetate- and zymosan-stimulated neutrophils. DMTU also provided complete inhibition when incubated with cell-free supernatants after the formation of the MPO products. DMTU prevented the oxidative inactivation of alpha 1-antitrypsin by neutrophil-stable oxidants. Evidence that DMTU was oxidized by the MPO products was obtained by titration of oxidized DMTU with reduced glutathione. Surprisingly, supernatants from cells incubated with DMTU (10 mM) consumed two- to threefold higher amounts of reduced glutathione than supernatants from cells incubated with taurine (15 mM). Metabolic studies with stimulated neutrophils and experiments with the MPO enzyme system in a cell-free system suggested that DMTU acts by scavenging the products of the MPO pathway rather than by blocking H2O2 production in the intact cell. These findings demonstrate that DMTU blocks the neutrophil MPO pathway in addition to its known ability to scavenge other reactive O2 species. The capacity of DMTU to scavenge MPO products may explain some of its protective effects in acute lung injury. PMID- 2551877 TI - Repeated antigen challenge induces airway hyperresponsiveness with tissue eosinophilia in guinea pigs. AB - To test the hypothesis that the development of airway hyperresponsiveness (AHR) lasting greater than or equal to 3 days after the last antigenic exposure required repeated mediator release, we compared dose-response changes in lung resistance (RL) to acetylcholine (ACh) in animals sensitized with 1% ovalbumin (OA), 4% Bordatella pertussis aerosol and subsequently challenged with 0.5% OA aerosol twice weekly for 4-6 wk vs. animals receiving saline aerosol instead of OA. Despite antihistamine pretreatment, each OA challenge produced cyanosis and inspiratory indrawing. Blood gas analysis in six guinea pigs revealed an immediate fall in arterial PO2 (PaO2) from 104.3 +/- 4.9 to 35.4 +/- 2.2 Torr after a 1-min exposure to aerosolized OA. ACh dose-response measurements of RL 3 days after the last OA challenge demonstrated a leftward shift and an increased magnitude of response. These differences were less marked at 7 days, and by 14 days after the last OA challenge, ACh dose-response curves were not different from those of control guinea pigs. Sensitization without repeated antigen challenge did not cause hyperresponsiveness. Morphometric analysis showed significantly increased numbers of eosinophils in the epithelium of airways in hyperresponsive guinea pigs, without neutrophil infiltration or alterations in epithelium and airway wall areas. We conclude that repeated antigenic challenge, but not sensitization alone, causes prolonged AHR in guinea pigs, which is associated with tissue eosinophilia. PMID- 2551878 TI - Exercise training increases coronary transport reserve in miniature swine. AB - Female yucatan miniature swine were trained on a treadmill (ET) or were cage confined (C) for 16-22 wk. The ET pigs had increased exercise tolerance, heart weight-to-body weight ratio, and skeletal muscle oxidative capacity. After anesthesia the left anterior descending coronary artery was cannulated and pump perfused with blood while aortic, central venous, and coronary perfusion pressures, electrocardiogram, heart rate, and coronary blood flow were monitored. Capillary permeability-surface area product (PS) for EDTA was determined with the single-injection indicator-diffusion method by use of an organ model based on the Sangren-Sheppard equations for capillary transport. Coronary blood flow (CBF) and PS were compared before and during maximal adenosine vasodilation with coronary perfusion pressures at 120 mmHg. Results indicate that there were no differences in base-line CBF or PS between C and ET groups. alpha-Receptor blockade with phentolamine and/or prazosin, before adenosine vasodilation, produced increases in PS in C pigs but had little effect in ET pigs. During maximal vasodilation with adenosine, ET pigs had greater CBF (447 +/- 24 vs. 366 +/- 27 ml.min-1.100 g 1) and greater PS (83 +/- 9 vs. 55 +/- 7 ml.min-1.100 g-1) than the C group. It is concluded that ET induces an increased coronary transport capacity in miniature swine that includes a 22% increase in blood flow capacity and a 51% increase in capillary exchange capacity. PMID- 2551879 TI - Calcitonin gene-related peptide vasodilation of human pulmonary vessels. AB - Human calcitonin gene-related peptide (CGRP) is localized to sensory neurons in pulmonary vessels and is a potent vasodilator. We have characterized the effects of CGRP in human pulmonary vessels and localized the receptors for this peptide by autoradiography. Fresh human lung tissue was obtained from eight patients undergoing surgery and small (200-400 microns ID) pulmonary arteries and veins were dissected free of surrounding connective and pulmonary tissue. Pairs of vessels were studied and in one of each pair the endothelium was left intact and from the other of each pair the endothelium was removed by gentle abrasion. For functional studies arteries (n = 9) and veins (n = 9) were suspended in an organ bath, precontracted with 1 microM prostaglandin F2 alpha. CGRP (10 pM to 10 microM) was added in a cumulative manner. CGRP caused a dose-dependent relaxation of endothelium intact human pulmonary arteries and veins with log EC50 values of 8.01 +/- 0.35 and -8.70 +/- 0.40, respectively (not significant). Removal of the endothelium did not diminish the vasodilator potency of CGRP in either vessel. For autoradiographic studies, cryostat sections of the small human pulmonary vessels with or without endothelium were used. 125I-CGRP densely labeled CGRP receptors on vascular smooth muscle and endothelial removal did not have any effect on grain density. We concluded that CGRP is a potent vasodilator of human pulmonary arteries and veins that is not dependent on an intact endothelium. These functional studies correlate with the distribution of CGRP receptors as localized by autoradiography. PMID- 2551880 TI - Effects of clonidine and dihydralazine on atrial natriuretic factor and cGMP in humans. AB - The effects of a 1-wk treatment with clonidine (75 micrograms/day twice a day) and dihydralazine (25 mg/day twice a day) on base-line levels of plasma atrial natriuretic factor (ANF) and plasma and urinary guanosine 3',5'-cyclic monophosphate (cGMP) and their changes by acute saline infusion (2 liters) in eight normal subjects were evaluated. Basal ANF was decreased to 65% in the clonidine group compared with both the control and dihydralazine groups. Volume loading increased plasma ANF levels by 30-40% of base-line values in the control and the dihydralazine groups and by 15% in the clonidine group. Basal plasma and urinary cGMP levels were raised by 30 and 90% in the dihydralazine group compared with both other groups. Volume loading increased plasma cGMP levels by 40% in the control and clonidine-treated groups and by 25% in the dihydralazine-treated group. It is concluded that ANF may contribute to hemodynamic effects of clonidine but not to those of dihydralazine. Dihydralazine increases plasma and urinary cGMP, supposedly by direct activation of the soluble guanylate cyclase. PMID- 2551881 TI - Culture of pancreatic islet cells and islet hormone producing cell lines "morphological and functional integrity in culture". PMID- 2551883 TI - Hepatocellular carcinoma with skeletal metastasis: management with intraarterial radioactive iodine. AB - Two patients with skeletal metastasis from hepatocellular carcinoma (HCC) treated with internal radiation are presented. An oily contrast medium injected into the hepatic artery accumulates in the hepatic tumor tissue and remains there for long periods: therefore, an oily contrast tagged with radioactive iodine could intensify therapeutic effect on tumors. I-131 was tagged onto Lipiodol, an iodized oil, and injected into the arteries supplying the tumors of skeletal metastasis and hepatocellular carcinoma delivering high internal radiation to both primary and secondary lesions. At 6-month follow-up period, tumor has decreased in size and patients are alive and pain free. PMID- 2551882 TI - Primary rabbit kidney proximal tubule cell cultures maintain differentiated functions when cultured in a hormonally defined serum-free medium. AB - A primary rabbit kidney epithelial cell culture system has been developed which retains differentiated functions of the renal proximal tubule. In addition, the cells have a distinctive metabolism and spectrum of hormone responses. The primary cells were observed to retain in vitro a Na+-dependent sugar transport system (distinctive of the proximal segment of the nephron) and a Na+-dependent phosphate transport system. Both of these transport processes are localized on the apical membrane of proximal tubule cells in vivo. In addition, probenicid sensitive p-aminohippurate (PAH) uptake was observed in basolateral membranes of the primary tubule cells, and the PAH uptake by these vesicles occurred at a rate that was very similar to that observed with membranes derived from the original tissue. Several other characteristics of the primary cells were examined, including hormone-sensitive cyclic AMP production and phosphoenolpyruvate carboxykinase (PEPCK) activity. Like the cells in vivo, the primary proximal tubule cells were observed to produce significant cyclic AMP in response to parathyroid hormone, but not in response to arginine vasopressin or salmon calcitonin. Significant PEPCK activity was observed in the particulate fraction derived from a homogenate of primary rabbit kidney proximal tubule cells. PMID- 2551884 TI - A facile and reversible method to decrease the copy number of the ColE1-related cloning vectors commonly used in Escherichia coli. AB - We report a technique which uses the cointegrate intermediate of transposon Tn1000 transposition as a means to lower the copy number of ColE1-type plasmids. The transposition of Tn1000 from one replicon to another is considered a two-step process. In the first step, the transposon-encoded TnpA protein mediates fusion of the two replicons to produce a cointegrate. In the second step, the cointegrate is resolved by site-specific recombination between the two transposon copies to yield the final transposition products: the target replicon with an integrated transposon plus the regenerated donor replicon. Using in vitro techniques, the DNA sequence of the Tn1000 transposon was altered so that cointegrate formation occurs but resolution by the site-specific recombination pathway is blocked. When this transposon was resident on an F factor-derived plasmid, a cointegrate was formed between a multicopy ColE1-type target plasmid and the conjugative F plasmid. Conjugational transfer of this cointegrate into a polA strain resulted in a stable cointegrate in which replication from the ColE1 plasmid origin was inhibited and replication proceeded only from the single-copy F factor replication origin. We assayed isogenic strains which harbored plasmids encoding chloramphenicol acetyltransferase to measure the copy number of such F factor-ColE1-type cointegrate plasmids and found that the copy number was decreased to the level of single-copy chromosomal elements. This method was used to study the effect of copy number on the expression of the fabA gene (which encodes the key fatty acid-biosynthetic enzyme beta-hydroxydecanoylthioester dehydrase) by the regulatory protein encoded by the fadR gene. PMID- 2551885 TI - Characterization of conjugal transfer functions of Agrobacterium tumefaciens Ti plasmid pTiC58. AB - Physical characterization of 13 transposon Tn5 insertions within the agrocinopine independent, transfer-constitutive Ti plasmid pTiC58Trac identified three separate loci essential for conjugation of this nopaline/agrocinopine A + B-type Ti plasmid. Complementation analysis with relevant subcloned DNAs indicated that the three physically separated blocks of conjugal genes constitute distinct complementation groups. Two independent Tn5 insertions within the wild-type, agrocinopine-dependent, repressed pTiC58 plasmid resulted in constitutive expression of conjugal transfer. These two insertions were physically indistinguishable and could not be complemented in trans. However, the Trac phenotype resulted when the Tn5-mutated fragment cointegrated into the wild-type Ti plasmid. While the spontaneous Trac mutant Ti plasmids were also derepressed for agrocinopine catabolism, those generated by Tn5 insertions remained inducible, indicating that this apparent cis-acting site is different from that affected in the spontaneous mutants. No chromosomal Tn5 insertion mutations were obtained that affected conjugal transfer. An octopine-type Ti plasmid, resident in different Agrobacterium tumefaciens chvB mutants, transferred at normal frequencies, demonstrating that this virulence locus affecting plant cell binding is not required for Ti plasmid conjugation. None of our conjugal mutants limited tumor development on Kalanchoe diagremontiana. Known lesions in pTiC58 vir loci had no effect on conjugal transfer of this Ti plasmid. These results show that pTiC58 Ti plasmid conjugal transfer occurs by functions independent of those required for transfer of DNA to plant cells. PMID- 2551886 TI - Cloning and characterization of a cluster of linked Bacillus subtilis late competence mutations. AB - We characterized a segment of chromosomal DNA from Bacillus subtilis that was required for the development of genetic competence. The chromosomal DNA was cloned from a group of genetically linked and phenotypically similar Tn917lac insertion mutants deficient in competence. This cluster of mutations defined the comG locus. Chromosomal DNA flanking each of the six insertions was cloned. Restriction maps of the cloned plasmids revealed that their chromosomal inserts consisted of overlapping fragments. These data, together with Southern blots of chromosomal DNA from the comG mutants, showed that the six Tn917lac comG insertions occurred in the following order: comG12, comG39, comG412, comG107, comG56, and comG210. Expression of the comG Tn917lac insertions was from a promoter located upstream from the first insertion, comG12. This was determined genetically and by low-resolution S1 nuclease mapping of the 3' terminus. The comG region spanned about 5 kilobase pairs, based on low-resolution S1 nuclease mapping of the transcription terminator and Northern blotting. The comG12 mutation had a partial epistatic effect on the expression of one other com locus, comE, but none of the other comG mutations affected expression of this or any other com gene tested. Based on these conclusions, and on its size and phenotype, the comG locus must be organized as a polycistronic operon that is subject to competence-specific regulation. PMID- 2551887 TI - Aromatic aminotransferase activity and indoleacetic acid production in Rhizobium meliloti. AB - Bacterial indoleacetic acid (IAA) production, which has been proposed to play a role in the Rhizobium-legume symbiosis, is a poorly understood process. Previous data have suggested that IAA biosynthesis in Rhizobium meliloti can occur through an indolepyruvate intermediate derived from tryptophan by an aminotransferase activity. To further examine this biosynthetic pathway, the aromatic aminotransferase (AAT) activity of Rhizobium meliloti 102F34 (F34) was characterized. At least four proteins were detected on nondenaturing gels of F34 protein extracts that exhibited AAT activity. All four of these AATs were constitutively produced and utilized the aromatic amino acids tryptophan, phenylalanine, and tyrosine as amino substrates. Two AATs were also capable of using aspartate. Plasmids from an F34 gene bank were identified that coded for the synthesis of at least three of these proteins, and the respective gene sequences were localized by transposon mutagenesis. Selected transposon insertions were recombined into the F34 genome to produce strains defective in two of these proteins (AAT1 and AAT2). Characterization of the mutants revealed that neither was essential for the biosynthesis of IAA in the absence of exogenous tryptophan, but that both contributed to IAA biosynthesis when high levels of exogenous tryptophan were present. AAT1 and AAT2 were also not required for the production of a minimal level of aromatic amino acids, but both were able to scavenge nitrogen from the aromatic amino acids during nitrogen deprivation. Neither AAT1 nor AAT2 was essential for symbiosis with alfalfa. PMID- 2551888 TI - Activation of a cryptic pathway for threonine metabolism via specific IS3 mediated alteration of promoter structure in Escherichia coli. AB - The tdh operon of Escherichia coli consists of two genes whose products catalyze sequential steps in the formation of glycine and acetyl coenzyme A from threonine. The operation of the tdh pathway can potentially confer at least two capabilities on the cell: the first is to provide a biosynthetic source of glycine, serine, or both that is an alternative to the conventional (triose phosphate) pathway; the second is to enable cells to utilize threonine as the sole carbon source. The latter capability is referred to as the Tuc+ phenotype. In wild-type E. coli, the tdh operon is expressed at levels that are too low to bestow the Tuc+ phenotype, even in leucine-supplemented media, where the operon is induced eightfold. In eight Tuc+ mutants, the expression of the tdh operon was elevated 100-fold relative to the uninduced wild-type operon. The physical state of the DNA at the tdh locus in these Tuc+ strains was analyzed by Southern blotting and by DNA sequencing. In eight independent isolates the mobile genetic element IS3 was found to lie within the tdh promoter region in identical orientations. In six cases that were examined by DNA sequencing, IS3 occupied identical sites between the -10 and -35 elements of the tdh promoter. The transcription start points for the wild-type tdh promoter and one IS3-activated tdh promoter were identical. In effect, the repeatedly observed transposition event juxtaposed an IS3-borne -35 region and the tdh-specific -10 region, generating a hybrid promoter whose utilization led to elevated, constitutive expression of the tdh operon. This is the first case of promoter activation by IS3 where the site of transcription initiation is unaltered. PMID- 2551889 TI - Use of phoA fusions to study the topology of the Escherichia coli inner membrane protein leader peptidase. AB - A topology of the Escherichia coli leader peptidase has been previously proposed on the basis of proteolytic studies. Here, a collection of alkaline phosphatase fusions to leader peptidase is described. Fusions to the periplasmic domain of this protein exhibit high alkaline phosphatase activity, while fusions to the cytoplasmic domain exhibit low activity. Elements within the cytoplasmic domain are necessary to stably anchor alkaline phosphatase in the cytoplasm. The amino terminal hydrophobic segment of leader peptidase acts as a weak export signal for alkaline phosphatase. However, when this segment is preceded by four lysines, it acts as a highly efficient export signal. The coherence of in vitro studies with alkaline phosphatase fusion analysis of the topology of leader peptidase further indicates the utility of this genetic approach to membrane protein structure and insertion. PMID- 2551890 TI - Identification and sequence analysis of the Rhizobium meliloti dctA gene encoding the C4-dicarboxylate carrier. AB - Transposon Tn5-induced C4-dicarboxylate transport mutants of Rhizobium meliloti 2011 which could be complemented by cosmid pRmSC121 were subdivided into two classes. Class I mutants (RMS37 and RMS938) were defective in symbiotic C4 dicarboxylate transport and in nitrogen fixation. They were mutated in the structural gene dctA, which codes for the C4-dicarboxylate carrier. Class II mutants (RMS11, RMS16, RMS17, RMS24, and RMS31) expressed reduced activity in symbiotic C4-dicarboxylate transport and in nitrogen fixation. These mutants were mutated in regulatory dct genes which do not play an essential role in the symbiotic state. Thin sections of alfalfa nodules induced by the wild type and class I and class II mutants were analyzed by light microscopy. Class mutants induced typical Fix- nodules, showing a large senescent zone, whereas nodules induced by class II mutants only differed in an enhanced content of starch granules compared with wild-type nodules. Class I mutants could be complemented by a 2.1-kilobase SalI-HindIII subfragment of cosmid pRmSC121. DNA sequencing of this fragment resulted in the identification of an open reading frame, which was designated dctA because Tn5 insertion sites of the class I mutants mapped within this coding region. The dctA gene was preceded by a nif consensus promoter and an upstream NifA-binding element. Upstream of the dctA promoter, the 5' end of the R. meliloti dctB gene could be localized. The amino acid sequence of the N terminal part of the R. meliloti DctB protein shared 49% homology with the corresponding part of the R. leguminosarum DctB protein. The DctA protein consisted of 441 or 453 amino acids due to two possible ATG start codons, with calculated molecular masses of 46.1 and 47.6 kilodaltons, respectively. The hydrophobicity plot suggests that DctA is a membrane protein with several membrane passages. The amino acid sequences of the R. meliloti and the R. leguminosarum DctA proteins were highly conserved (82%). PMID- 2551892 TI - Tn5 mutagenesis of Anabaena sp. strain PCC 7120: isolation of a new mutant unable to grow without combined nitrogen. AB - Methylation by Ava methylases in Escherichia coli increases the efficiency to transfer of Tn5 in pBR322bla:: Tn5 from E. coli to Anabaena sp. strain PCC 7120 by conjugation. Following conjugation, Tn5 but not pBR322 sequences were found at many different positions in the Anabaena chromosome. This procedure was used to mutagenize, tag, and clone a previously unrecognized gene required for nitrogen fixation in this Anabaena sp. PMID- 2551891 TI - Isolation and characterization of mutants with deletions in dnaQ, the gene for the editing subunit of DNA polymerase III in Salmonella typhimurium. AB - dnaQ (mutD) encodes the editing exonuclease subunit (epsilon) of DNA polymerase III. Previously described mutations in dnaQ include dominant and recessive mutator alleles as well as leaky temperature-sensitive alleles. We describe the properties of strains bearing null mutations (deletion-substitution alleles) of this gene. Null mutants exhibited a growth defect as well as elevated spontaneous mutation. As a consequence of the poor growth of dnaQ mutants and their high mutation rate, these strains were replaced within single colonies by derivatives carrying an extragenic suppressor mutation that compensated the growth defect but apparently not the mutator effect. Sixteen independently derived suppressors mapped in the vicinity of dnaE, the gene for the polymerization subunit (alpha) of DNA polymerase III, and one suppressor that was sequenced encoded an altered alpha polypeptide. Partially purified DNA polymerase III containing this altered alpha subunit was active in polymerization assays. In addition to their dependence on a suppressor mutation affecting alpha, dnaQ mutants strictly required DNA polymerase I for viability. We argue from these data that in the absence of epsilon, DNA replication falters unless secondary mechanisms, including genetically coded alteration in the intrinsic replication capacity of alpha and increased use of DNA polymerase I, come into play. Thus, epsilon plays a role in DNA replication distinct from its known role in controlling spontaneous mutation frequency. PMID- 2551893 TI - Purification and characterization of a growth factor from guinea pig harderian gland. AB - A polypeptide growth factor, Harderian gland-derived growth factor (HGDGF), has been purified approximately 43,000-fold from guinea pig Harderian gland by column chromatography on TSK gel DEAE-5PW, blue-Sepharose CL-6B, and Superose 12. The yield was approximately 10%. The Superose 12 fraction was further purified by Aquapore BU-300 reversed-phase chromatography to apparent homogeneity. HGDGF was eluted from TSK gel DEAE-5PW at 0.20-0.35 M NaCl, with a linear gradient of 0.15 0.80 M NaCl and at 2.20 M NaCl from blue-Sepharose CL-6B. The activity of HGDGF toward human embryonic cells (TIG-3) was quantitated, [3H]thymidine incorporation for 48 h being stimulated in a linear and dose-dependent manner. Purified HGDGF has a molecular weight of approximately 13,000, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and molecular sieve column chromatography. HGDGF is labile to treatment with SH reagents or acetic acid. Both trypsin digestion and boiling decrease the activity of HGDGF. Its pI is 5.1. HGDGF stimulates the multiplication of TIG-3 cells but has no effect on human endothelial cells K2T1 or A2T2 which require fibroblast growth factor for growth. HGDGF appears to differ from other growth factors, suggesting that it is a previously undescribed growth factor. PMID- 2551894 TI - Synthesis of peptides as cloned ubiquitin extensions. AB - Oligonucleotides encoding four peptides ranging in length from 10 to 21 amino acids were cloned between the Afl2 and KpnI sites in the ubiquitin expression vector, pNMHUb. Escherichia coli AR13, a strain that contains a temperature sensitive lambda repressor, was transformed by the plasmids, and upon shift to 42 degrees C, the cells produced large amounts of ubiquitin extended at its carboxyl terminus by each of the four peptides. Following a simple three-step purification of the ubiquitin fusion proteins, the peptides were cleaved from ubiquitin using a ubiquitin-alpha-protein hydrolase isolated from rabbit reticulocytes. The released ubiquitin molecules were shown to be competent in conjugation assays, and amino acid analyses of the purified peptides revealed that full length products were obtained in all cases. Since peptide yields varied from 2 to 4 mg/liter of culture medium, ubiquitin extensions provide an attractive alternative to solid phase synthesis for the production of peptides. PMID- 2551895 TI - Molecular determinants of receptor affinity and selectivity of the natural delta opioid agonist, dermenkephalin. AB - Processing of the polyprotein precursor pro-dermorphin generates two distantly related D-amino acid-containing peptides, dermorphin and dermenkephalin, which are among the most selective high affinity agonists described, respectively, for the mu- and delta-opioid receptors. Dermenkephalin, Tyr-D-Met-Phe-His-Leu-Met-Asp NH2, is a linear, potentially flexible peptide devoid of structural homology with either enkephalins, endorphins, or dynorphins and, as such, represents a useful tool for identifying determinants of high affinity and selective binding of opioids to the delta-receptor. A series of selected dermenkephalin analogs and homologs was investigated for affinity at the mu- and delta-sites in the brain. Whereas dermenkephalin has high affinity and specificity for the delta-opioid receptors, its tetrapeptide amino end, dermenkephalin-[1-4]-NH2 binds almost exclusively at the mu-receptors. Dermorphin, Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH2, is only marginally more selective for the u-sites than is dermenkephalin-[1-4] NH2. Using dermorphin-dermenkephalin peptide hybrids and C-terminal deletion analogs of dermenkephalin, we showed the critical role that the C-terminal residues Met6 and Asp7 play in specifying correct addressing of dermenkephalin toward delta-receptors. The potent mu-deteminant located within the amino end of dermenkephalin is over-whelmed by the powerful delta-directing ability of the carboxy end. The negatively charged side chain of Asp7 makes a significant contribution to the delta-addressing ability of the C-terminal region, a finding consistent with Schwyzer's membrane selection model (Schwyzer, R. (1986) Biochemistry 25, 6335-6342). The Leu residue in position 5 and D-configuration about the alpha-carbon of Met2 were found to be of crucial importance for high affinity binding to delta-receptors. Whereas the Met residue in position 6 in dermenkephalin could safely be oxidized or replaced with D-Met, oxidation of Met2 led to deleterious effects, this analog being 1/100 as potent as dermenkephalin at delta-sites. Overall, the data collected demonstrate that highest levels of selectivity and affinity for the delta-opioid receptors can be achieved with small-sized, potentially flexible, linear peptides and further support the model according to which, in addition to optimum accommodation at the receptor, selection for delta-receptors is reduced by the effective positive charge of the molecule. Dermenkephalin may provide a starting point for the design of agonists and antagonists with nearly total specificity for the delta-sites. Such pharmacological agents could be used to explore the ill-defined physiological role and behavioral actions conveyed by delta-opioid receptors. PMID- 2551896 TI - Lateral mobility of both envelope proteins (F and HN) of Sendai virus in the cell membrane is essential for cell-cell fusion. AB - Fluorescence photobleaching recovery was employed to study the effects of specific immobilization of Sendai virus envelope glycoproteins (F, the fusion protein, and HN, the hemagglutinin-neuraminidase) on the virally mediated fusion of human erythrocytes. Lateral immobilization of varying fractions of F and/or HN (after virus adsorption and hemagglutination, but before fusion) was achieved by cross-linking them with succinyl concanavalin A (inhibiting both F and HN) or with specific rabbit IgG directed against either F or HN. Alternatively, agglutinated cells were treated with low concentrations of the above proteins (inducing only minor inhibition of either mobility or fusion), and immobilization of F and/or HN was induced by cross-linking with a secondary antibody; this protocol ensured a minimal contribution of direct binding to the viral proteins to the inhibition of fusion. Our results demonstrate that lateral immobilization of either F or HN results in a strong inhibition of cell-cell fusion and a much weaker inhibition of virus-cell fusion. The level of cell-cell fusion was directly correlated with the level of laterally mobile viral glycoproteins in the cell membrane (either F or HN). We conclude that lateral mobility of both F and HN in the red cell membrane is essential for cell-cell fusion and that not only F but also HN has a role in this fusion event. The possible reasons for the different dependence of cell-cell and virus-cell fusion on viral glycoprotein mobility are discussed. PMID- 2551897 TI - A bisubstrate analog inhibitor for alpha(1----2)-fucosyltransferase. AB - Porcine submaxillary beta-galactoside alpha(1----2)-fucosyltransferase is known to transfer a fucosyl residue from guanosine 5'-diphosphofucose (GDP-fucose) to the 2-OH group of beta-D-galactopyranosides with inversion of configuration at the fucopyranosyl anomeric carbon. A bisubstrate analog (1) of the postulated transition-state for this reaction, which has O-2 of phenyl beta-D galactopyranoside attached to the terminal phosphorous of GDP through a flexible ethylene bridge, has been chemically synthesized and evaluated as an inhibitor of this enzyme. Compound 1 was found to be a competitive inhibitor with respect to both GDP-fucose and phenyl beta-D-galactopyranoside for both the membrane-bound and soluble forms of the fucosyltransferase. It was also a competitive inhibitor with respect to the alternate acceptor beta DGal(1----3)beta DGlcNAcO(CH2)8 COOMe. The Ki values were in the range 2.3-16 microM. Compound 1 is the first example of a bisubstrate analog inhibitor for a glycosyltransferase which binds to both the acceptor and donor recognition sites of the enzyme. The potential of a bisubstrate analog strategy for the production of specific glycosyltransferase inhibitors is discussed. PMID- 2551899 TI - Role of tryptophan 26 in the NAD glycohydrolase reaction of the S-1 subunit of pertussis toxin. AB - The gene encoding a catalytically active deletion peptide, the C180 peptide, of the S-1 subunit of pertussis toxin was engineered to facilitate mutagenesis at the Trp-26 (wild-type) coding sequence. A synthetic double-stranded oligonucleotide was inserted into the C180 gene such that all possible codons would be introduced into position 26. Seven individual mutants of the C180 peptide which possessed amino acid substitutions at residue 26 (collectively termed C180W26n peptides) were purified from periplasmic extracts of Escherichia coli. Each C180W26n peptide was present as a single major peptide that had an apparent molecular mass of between 20.9 and 24.5 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and each showed similar immunoreactivity relative to the C180 peptide. The C180W26n peptides demonstrated marked reduction of both ADP-ribosyltransferase and NAD glycohydrolase activities at 25 nM and 10 microM NAD, respectively. Kinetic analysis of the two most active mutants, C180W26F and C180W26Y, revealed that the major perturbation of NAD glycohydrolase activity was due to an increase (approximately 20-fold) in the Km for NAD between these mutants and the C180 peptide. PMID- 2551898 TI - SV40-transformed human lung fibroblasts secrete a 92-kDa type IV collagenase which is identical to that secreted by normal human macrophages. AB - We have reported that SV40-transformed human lung fibroblasts secrete a 92-kDa metalloprotease which is not detectable in the parental cell line IMR-90. We now present the complete structure of this enzyme along with the evidence that it is identical to the 92-kDa metalloprotease secreted by normal human alveolar macrophages, phorbol ester-differentiated monocytic leukemia U937 cells, fibrosarcoma HT1080 cells, and cultured human keratinocytes. A similar, perhaps identical, enzyme can be released by polymorphonuclear cells. The preproenzyme is synthesized as a polypeptide of predicted Mr 78,426 containing a 19 amino-acid long signal peptide and secreted as a single 92,000 glycosylated proenzyme. The purified proenzyme complexes noncovalently with the tissue inhibitor of metalloproteases (TIMP) and can be activated by organomercurials. Activation with phenylmercuric chloride results in removal of 73 amino acids from the NH2 terminus of the proenzyme, yielding an active form capable of digesting native types IV and V collagen. The in vitro substrate specificity of the enzyme using these substrates was indistinguishable from that of the 72-kDa type IV collagenase. The 92-kDa type IV collagenase consists of five domains; the amino terminal and zinc-binding domains shared by all members of the secreted metalloprotease gene family, the collagen-binding fibronectin-like domain also present in the 72-kDa type IV collagenase, a carboxyl-terminal hemopexin-like domain shared by all known enzymes of this family with the exception of PUMP-1, and a unique 54-amino-acid-long proline-rich domain homologous to the alpha 2 chain of type V collagen. PMID- 2551900 TI - Calmodulin regulates fodrin susceptibility to cleavage by calcium-dependent protease I. AB - The intracellular calcium-dependent proteolysis of fodrin has been postulated to be central to the regulation of plasticity of the cortical cytoskeleton of many eukaryotic cells. The close proximity of the sites of calmodulin (CaM) binding and calcium-dependent protease I (CDP-I) cleavage in mammalian alpha-fodrin suggested that their action may be linked. In hypotonic and isotonic buffers, CDP I proteolysis of the beta subunit of fodrin was absolutely dependent upon the presence of active CaM. The stimulation by CaM was inhibited by CaM antagonists. The rate of CDP-I proteolysis of both subunits was enhanced by CaM, while the rate of fodrin proteolysis with other proteases was not influenced by CaM. The increase in the susceptibility of fodrin to CDP-I proteolysis was half-maximal at 80 nM CaM, and maximal at 200 nM CaM. The unusual and differential susceptibility of alpha- and beta-fodrin to proteolysis by CDP-I in the absence of CaM was exploited to investigate the quaternary structure of fodrin in which only the alpha subunit was cleaved. Cleavage of the alpha subunit alone did not destroy the tetrameric form of the molecule, whereas CDP-I cleavage of both subunits rendered the molecule incapable of reforming tetramers. These results provide structural and functional evidence that CaM and CDP-I act synergistically in the regulated proteolysis of fodrin. PMID- 2551901 TI - An ATP-driven Cl- pump in the brain. AB - EDTA-treated microsomes prepared from rat brain mainly consisted of sealed membrane vesicles 200-500 nm in diameter and were rich in both Cl- -ATPase and Na+,K+-ATPase activities. Such Cl- -ATPase-rich membrane vesicles accumulated Cl- in an ATP-dependent and osmotically reactive manner in the presence of 1 nM ouabain. The Cl- uptake was maximally stimulated by ATP with a Km value of 1.5 mM; GTP, ITP, and UTP partially stimulated Cl- uptake, but CTP, beta, gamma methylene ATP, ADP, and AMP did not. The ATP-dependent Cl- uptake was accelerated by an increase in the medium Cl- concentration with a Km value of 7.4 mM. Such stimulation of Cl- uptake by ATP was dependent on the pH of the medium, with an optimal pH of 7.4, and also on the temperature of the medium, with an optimal range of 37-42 degrees C. Ethacrynic acid dose dependently inhibited the ATP dependent Cl- uptake with a concentration for half-maximal inhibition at 57 microM. N-ethylmaleimide (0.1 mM) completely inhibited and sodium vanadate (1 mM) partially inhibited the ATP-dependent Cl- uptake. The membrane vesicles did not accumulate H+ in the Cl- uptake assay medium. The ATP-dependent Cl- uptake profile agreed with that of Cl- -ATPase activity reported previously (Inagaki, C., Tanaka, T., Hara, M., and Ishiko, J. (1985) Biochem. Pharmacol. 34, 1705 1712), and this strongly supports the idea that Cl- -ATPase in the brain actively transports Cl-. PMID- 2551902 TI - Identification of major autolytic cleavage sites in the regulatory subunit of vascular calpain II. A comparison of partial amino-terminal sequences to deduced sequence from complementary DNA. AB - Purified calpain II from vascular smooth muscle is a heterodimer consisting of catalytic (Mr = 76,000) and regulatory (Mr = 30,000) subunits. In the presence of Ca2+, the regulatory subunit undergoes stepwise autolysis resulting in enzyme activation. By slowing autoproteolysis, we identified major autolytic intermediates of the regulatory subunit. Gas-phase sequencing of the regulatory subunit and its autolytic fragments revealed that the NH2-terminus of the Mr = 30,000 form was blocked, whereas each fragment yielded a unique amino acid sequence, suggesting that autolysis proceeds in an NH2- to COOH-terminal direction. By comparison of actual amino acid sequences of autolytic cleavage intermediates to the full sequence deduced from cDNA, we have identified the major autolytic cleavage sites. Three different peptide bonds were cleaved, with neutral amino acids predominating on both sides of the peptide bond hydrolyzed. Importantly, leucine or isoleucine was identified in the second position upstream from the cleavage site in all three autolytic sequences. The presence of an upstream leucine residue in the autolytic cleavage sequence is reminiscent of the structure of potent microbial and synthetic peptide inhibitors of calpain. PMID- 2551903 TI - Purification and characterization of two related but distinct metalloproteinase inhibitors secreted by bovine aortic endothelial cells. AB - Two metalloproteinase inhibitors were purified from serum-free medium conditioned by bovine aortic endothelial cells. One of these inhibitors, with a molecular weight of 30,000-34,000 (reduced) is identified as tissue inhibitor of metalloproteinases; the second inhibitor has a molecular weight of 27,500 (reduced) and 20,400 (unreduced), is not recognized by an antiserum against bovine tissue inhibitor of metalloproteinases, appears unglycosylated, and has 51% identity with tissue inhibitor of metalloproteinases by NH2-terminal amino acid sequence analysis. This inhibitor has antiproteinase activities similar to those of tissue inhibitor of metalloproteinases, with inhibition of classical collagenase, type IV collagenase, and gelatinases but not trypsin, plasmin, or bacterial collagenase. Other properties shared with tissue inhibitor of metalloproteinases include trypsin sensitivity, acid and heat resistance, and inactivation by reduction-alkylation. The presence of these inhibitors in endothelial cells suggests that they may play important roles in protecting the integrity of the vascular basement membrane. PMID- 2551904 TI - Effect of hormones and development on the expression of the rat 1,25 dihydroxyvitamin D3 receptor gene. Comparison with calbindin gene expression. AB - We have used specific cDNAs to the rat vitamin D receptor (VDR) and to the mammalian vitamin D-dependent calcium-binding proteins (calbindin-D9k in intestine and calbindin-D28k in kidney) in order to obtain a better understanding of the regulation of the VDR gene and its relationship to calbindin gene expression. Hormonal regulation and development expression of the rat VDR gene were characterized by both Northern and slot blot analyses. Administration of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3; 25 ng/day for 7 days) to vitamin D deficient rats resulted in an increase in calbindin mRNA in intestine and kidney but no change in VDR mRNA in these tissues. Vitamin D-deficient rats responded to dexamethasone treatment (100 micrograms/100 g of body weight/day for 4 days) with a 2.5-fold increase in intestinal VDR mRNA which was accompanied by a 4-fold decrease in intestinal calbindin-D9k mRNA. Developmental studies indicated a pronounced increase in renal VDR mRNA and calbindin-D28k mRNA between birth and 1 week of age. In the intestine, an induction of VDR and calbindin-D9k gene expression was observed at a later time, during the 3rd postnatal week (the period of increased duodenal active transport of calcium). Taken collectively, our data indicate that in the adult rat, target tissue response to hormone is not modified by a corresponding alteration in new receptor synthesis. However, developmental studies indicate that the induction of 1,25(OH)2D3 receptor mRNA is correlated with the induction of calbindin gene expression. Our results also demonstrate that glucocorticoid administration can result in an alteration in intestinal calbindin and VDR gene expression. PMID- 2551905 TI - The structure of tumor necrosis factor-alpha at 2.6 A resolution. Implications for receptor binding. AB - The three-dimensional structure of tumor necrosis factor (TNF-alpha), a protein hormone secreted by macrophages, has been determined at 2.6 A resolution by x-ray crystallography. Phases were determined by multiple isomorphous replacement using data collected from five heavy atom derivatives. The multiple isomorphous replacement phases were further improved by real space symmetry averaging, exploiting the noncrystallographic 3-fold symmetry of the TNF-alpha trimer. An atomic model corresponding to the known amino acid sequence of TNF-alpha was readily built into the electron density map calculated with these improved phases. The 17,350-dalton monomer forms an elongated, antiparallel beta-pleated sheet sandwich with a "jelly-roll" topology. Three monomers associate intimately about a 3-fold axis of symmetry to form a compact bell-shaped trimer. Examination of the model and comparison to known protein structures reveals striking structural homology to several viral coat proteins, particularly satellite tobacco necrosis virus. Locations of residues conserved between TNF-alpha and lymphotoxin (TNF-beta, a related cytokine known to bind to the same receptors as TNF-alpha) suggest that lymphotoxin, like TNF-alpha, binds to the receptor as a trimer and that the general site of interaction with the receptor is at the "base" of the trimer. PMID- 2551906 TI - Assay sensitivity and differentiation of monoclonal antibody specificity in ELISA with different coating buffers. AB - Buffers of different pH and ionic strength were employed as coating buffers for antigen adsorption to microtitre plates. Their efficiency for coating plates with rinderpest virus (RPV) and foot-and-mouth disease virus (FMDV) antigens was studied by ELISA with polyclonal and monoclonal antibody preparations. While the adsorption and detection of RPV antigen with polyclonal antiserum was highly dependent on the ionic strength and pH of coating buffer, adsorption of antigenically active FMDV antigen was relatively unaffected by the buffering conditions. Both antigens were adsorbed optimally in 0.01 M phosphate buffer, pH 8.0. When monoclonal antibodies were used to detect antigen, there was a greater degree of dependence on the coating buffer than that found with polyclonal antisera. Moreover, when they were used to detect antigen adsorbed under several buffering conditions, monoclonal antibodies showed a variety of preferred buffers. The usefulness of this differential reactivity in distinguishing epitope specificity is demonstrated. PMID- 2551907 TI - Control of endotoxemia in burn patients by use of polymyxin B. AB - A group of patients with severe burns were entered into two sequential prospective randomized trials for reduction of endotoxemia by the use of intravenous polymyxin B. The first group underwent polymyxin administration during the first week after burn injury in a bell-shaped dosage form constructed to resemble the level of endotoxemia as previously documented. This group showed a statistically highly significant reduction in endotoxin levels and a suggestive, but not statistically significant, reduction in wound infection and mortality in the treated group compared with controls. The second group of patients underwent treatment with perioperative polymyxin B given in conjunction with an excisional procedure of the burn wound. In this group, polymyxin B also accomplished a reduction in endotoxemia from preoperative to postoperative cases, but there was no significant reduction in clinical complication rate or mortality. In the dosages used, polymyxin B is nontoxic and promises to be a useful part of the surgeon's armamentarium in dealing with severe complications of gram-negative sepsis. PMID- 2551908 TI - Na+,K+-adenosine triphosphatase polarity in retinal photoreceptors: a role for cytoskeletal attachments. AB - We have used isolated embryonic photoreceptor cells as a model system with which to examine the mechanisms responsible for the development and maintenance of asymmetric Na+,K+-ATPase (ATPase) distribution. Photoreceptor precursors, which appear round and process free at culture onset, develop structural and molecular properties similar to those of photoreceptor cells in vivo. ATPase, recognized by an anti-ATPase antibody, is distributed over the entire surface of round photoreceptor precursors. As the cells develop, ATPase becomes progressively concentrated in the inner segment (where it is found in cells of the intact retina). This phenomenon occurs in cells developing in the absence of intercellular contacts. The development of ATPase polarity correlates with a decrease in the fraction of ATPase molecules that are mobile in the membrane (as determined by fluorescence photobleaching recovery), as well as with an increase in the fraction of ATPase that remains associated with the cells after detergent extraction. The magnitudes of the mobile ATPase fractions agree well with those of the detergent-extractable fractions in both the immature and developed photoreceptors. The distribution of alpha spectrin and ATPase-immunoreactive materials appeared qualitatively similar, and quantitative image analysis showed similar gradients of spectrin and Na+,K+-ATPase immunofluorescence along the long axis of elongated photoreceptors. Moreover, detergent extractability of alpha spectrin and the ATPase showed similar modifications in response to changes in pH or KCl concentration. ATPase detergent-extractable and mobile fractions were not changed in cultures treated with cytoskeletal inhibitors such as nocodazole. These data are consistent with a role for an asymmetrically distributed, spectrin containing subcortical cytoskeleton in the preferential accumulation of Na+,K+ ATPase in the photoreceptor inner segment. PMID- 2551909 TI - Wheat germ agglutinin induces mating reactions in Chlamydomonas eugametos by cross-linking agglutinin-associated glycoproteins in the flagellar membrane. AB - Species-specific binding between the flagellar surfaces of mating types plus and minus (mt+ and mt-) gametes of Chlamydomonas eugametos is mediated by mating type specific agglutinins. Their interaction triggers several mating responses that are necessary for cell fusion, such as flagellar twitching, flagellar tip activation, redistribution of agglutinin molecules to the flagellar tip (tipping), and mating structure activation. Earlier, we reported that a monoclonal antibody (mAb 66.3) can induce mating reactions by cross-linking the agglutinins (Homan, W. L., A. Musgrave, H. de Nobel, R. Wagter, A. H. J. Kolk, D. de Wit, and H. van den Ende. 1988. J. Cell Biol. 107:177-189). Here we report that the lectin wheat germ agglutinin (WGA), which does not bind to the agglutinins, can also invoke all these mating reactions. We show, by immunofluorescence studies using anti-WGA and an agglutinin-specific monoclonal antibody (mAb 66.3), that WGA induces the redistribution of agglutinin to the flagellar tips of mt- gametes. Vice versa, when agglutinin tipping is induced by mAb 66.3, the WGA-binding glycoproteins are also tipped. Under the same conditions, the major flagellar glycoproteins are not redistributed, indicating that membrane transport is limited to a few components. We conclude that each agglutinin is associated with a WGA-binding glycoprotein. When cells lacking agglutinin or cells possessing inactive agglutinins are treated with WGA, mating responses are again elicited. The data suggest that clustering of agglutinin containing complexes results in the production of intracellular signals, such as cAMP, and the coupling of the complex to a force generating system. In nature, the complexes are clustered via the agglutinins, but artificially they can be clustered by lectins or antibodies directed against other proteins in the complex. PMID- 2551911 TI - The mitogenic signaling pathway but not the plasminogen activator-inducing pathway of basic fibroblast growth factor is mediated through protein kinase C in fetal bovine aortic endothelial cells. AB - Basic fibroblast growth factor (bFGF) induces cell proliferation and plasminogen activator (PA) activity in transformed fetal bovine aortic endothelial (FBAE) GM 7373 cells. A similar response is observed after treatment with 12-O tetradecanoylphorbol-13-acetate (TPA). In these cells, bFGF and TPA cause activation of protein kinase C (PKC), as demonstrated by the induction of the phosphorylation of an 87-kD PKC substrate in intact cells and by the increase in membrane-associated PKC activity. Activation of PKC by bFGF or TPA is inhibited in cells made PKC-deficient by pretreatment with high concentrations of TPA. The mitogenic activity of bFGF or of TPA is completely inhibited in PKC-deficient cells or in naive cells treated with the PKC inhibitor H-7. However, these cells proliferate in response to serum, epidermal growth factor, and dibutyryl cyclic AMP. Similar results are obtained in normal FBAE AG 7680 cells. These data indicate that activation of PKC is responsible for the mitogenic activity of bFGF in FBAE cells. On the contrary, the PA-inducing activity of bFGF is unaffected by down-regulation of PKC or by treatment with the PKC inhibitor H-7 in both transformed GM 7373 and normal AG 7680 cells. bFGF induces a rapid 45Ca influx in naive and in PKC-deprived GM 7373 cells. In these cells, addition of EGTA to the incubation medium prevents both the 45Ca influx and the increase in PA activity induced by bFGF, without affecting its mitogenic activity. Even though the involvement of PKC in the increase of cell-associated PA activity induced by bFGF can not be completely dismissed, the present results suggest a role of calcium entry in the modulation of the PA-inducing activity of bFGF. PMID- 2551910 TI - Regulated secretion of a serine protease that activates an extracellular matrix degrading metalloprotease during fertilization in Chlamydomonas. AB - During fertilization in the biflagellated alga, Chlamydomonas reinhardtii, gametes of opposite mating types adhere to each other via agglutinin molecules located on their flagellar surfaces, generating a sexual signal that induces several cellular responses including cell wall release. This cell contact generated signal is mediated by cAMP and release of the wall, which is devoid of cellulose and contains several hydroxyproline-rich glycoproteins, is due to the activation of a metalloprotease, lysin. Although we originally assumed that lysin would be stored intracellularly in a compartment structurally separate from its substrate, recently we showed that lysin is stored in the periplasm as an inactive, higher relative molecular mass precursor, prolysin (Buchanan, M.J., S. H. Imam, W. A. Eskue, and W. J. Snell. 1989. J. Cell Biol. 108:199-207). Here we show that conversion of prolysin to lysin is due to a cellular, nonperiplasmic enzyme that has the properties of a serine protease. Release of this serine protease into the periplasm is induced by incubation of gametes in dibutyryl cAMP. This may be one of the few examples of regulated secretion of a protease in a eucaryotic microorganism and a novel example of regulated secretion in a plant system. PMID- 2551912 TI - Genetically engineered (chimeric) antibodies. AB - The diversity, specificity, and biologic activity of antibodies make them potentially ideal reagents in the clinic and the laboratory, but conventional antibodies do not always fill the bill. Naturally occurring antibodies are heterogeneous, and human hybridoma antibodies are hard to produce. Genetic engineering techniques offer antibodies with just the properties desired. PMID- 2551913 TI - Managing patients after cardiac transplantation. AB - To care for heart transplant recipients is to walk an endless tightrope, teetering between too little immunosuppression, and consequent rejection episodes, and too much immunosuppression, with its correlated infection and neoplasia risks. Moreover, as long-term survival has become a reality, accelerated coronary vascular disease poses a major threat to the transplanted organ. PMID- 2551914 TI - Reversed-phase liquid chromatographic determination of 2',3'-dideoxycytidine in human blood samples. PMID- 2551915 TI - Adrenocorticotropin stimulation of aldosterone: prolonged continuous versus pulsatile infusion. AB - Continuous iv administration of ACTH leads to a sustained stimulation of cortisol but a transient stimulation of aldosterone followed by a decline to prestimulation levels by 72 h. Since CRH and ACTH are released in a pulsatile pattern in man, this study sought to investigate whether pulsatile administration of alpha-cosyntropin-(1-24) would lead to the maintenance of aldosterone stimulation over time. Eight normal male subjects on a 10-meq sodium, 100-meq potassium diet received both a continuous and a pulsatile (0.33 U ACTH/pulse over 15 min, pulsed every 2 h) infusion of cosyntropin (4 U/24 h) for 48 h (n = 4) or 72 h (n = 4). Aldosterone and cortisol were sampled every 6 h, and PRA and angiotensin-II every 24 h. Continuous infusion led to a stimulation of aldosterone followed by a progressive decline to preinfusion levels by 72 h [preinfusion 29 +/- 5 ng/dL (810 +/- 139 pmol/L); 72 h, 38 +/- 10 ng/dL (1054 +/- 277 pmol/L); P = 0.40]. Pulsatile infusion led to a stimulation of aldosterone which was maintained up to 72 h [preinfusion 33 +/- 7 ng/dL (915 +/- 194 pmol/L); 72 h, 85 +/- 13 ng/dL (2358 +/- 361 pmol/L); P less than 0.05]. Regression analysis of aldosterone (y) over time (x) from the peak level at 18 h for the continuous infusion showed a significant negative relation (r = 0.63; P = 0.001), indicating a progressive decline in aldosterone. However, for the pulsatile infusion, there was no relation (r = 0.02; P = 0.85), indicating maintenance of aldosterone levels. There were no significant differences in sodium, potassium, PRA, angiotensin-II, or cortisol between infusions to explain these differences in aldosterone levels. Therefore, pulsatile infusion of cosyntropin maintains aldosterone secretion over time. PMID- 2551916 TI - Delirium and neuromuscular symptoms in an elderly man with isolated corticotroph deficiency syndrome completely reversed with glucocorticoid replacement. AB - A 68-yr-old man had developed intractable vomiting soon after recovering from a flu-like illness. The use of Compazine as an antiemetic produced classic dystonic manifestations which resolved rapidly after discontinuation and treatment with Artane. However, he later developed a variety of neurobehavioral disturbances which led to his admission to the hospital. Extensive diagnostic procedures failed to identify any gastrointestinal or neurological causes. His condition unceasingly worsened until hypocortisolemia was serendipitously discovered, and all of his symptoms disappeared rapidly and completely with glucocorticoid replacement. Over the course of hospitalization, other than a single episode of orthostatic hypotension, the patient did not manifest any signs of adrenal insufficiency or endocrinopathy. Although detectable, his plasma ACTH level was markedly low in the presence of hypocortisolemia. His adrenal function was subnormal in the cortisol response to ACTH stimulation. His renin-angiotensin aldosterone system and catecholamine levels were normal. He had normal pituitary responses to GnRH, TRH, and insulin, with rises in plasma levels of LH, FSH, TSH, PRL, and GH, but no stimulation of ACTH. Repeated CRH tests revealed no stimulation of ACTH and cortisol. No circulating anti-ACTH, antiadrenal, or antipituitary antibody was detected. We conclude that this elderly patient had a rare syndrome of selective corticotroph dysfunction which resulted in secondary adrenal failure and exacerbated his mental and neuromuscular abnormalities. To our knowledge, these symptoms, which clearly relate to hypocortisolism, have not been previously reported. PMID- 2551917 TI - Insulin-like growth factor secretion by human B-lymphocytes: a comparison of cells from normal and pygmy subjects. AB - Freshly isolated human B-lymphocytes from eight subjects and Epstein-Barr virus transformed human B-lymphocytes from seven subjects were examined for their capacity to secrete insulin-like growth factor-I (IGF-I) and IGF-II and for their capacity to respond to human GH. Similar studies were conducted with Epstein-Barr virus-transformed lymphocytes collected from six African pygmies. When transformed B-lymphocytes from normal stature subjects were cultured for 3 weeks in RPMI-1640 medium (6 x 10(3) cells/75-cm2 flask at seeding), significant amounts of IGF-I, but no IGF-II, were produced. GH (150 ng/mL) significantly increased for control cells the amount of IGF-I produced at each sampling interval compared to that by unstimulated cultures (P less than 0.05 at 1 week; P = 0.005 at 3 weeks). At 3 weeks, cell counts of cultures compared were 4.13 +/- 0.39 X 10(6)/mL for unstimulated cells and 4.23 +/- 0.87 X 10(6)/mL for GH stimulated cells. IGF-I production at this time interval by unstimulated cells was 2.8 +/- 2.3 ng/mL, and that by GH-stimulated cells was 12.3 +/- 2.5 ng/mL (P = 0.005). Cell multiplication rates of control cultures were increased in 1 week by GH stimulation [GH stimulated, [16.7 +/- 22.0 X 10(4) cells, unstimulated, 5.73 +/- 4.1 X 10(4) cells; (mean +/- SD); n = 14; P less than 0.01]. Similar results occurred with GH studied at a lower concentration of 10 ng/mL for 3 weeks. Freshly isolated B-lymphocytes did not secrete IGF-I and II after 5 days of culture with GH. Cultures established from cells derived from pygmies produced significantly less IGF-I (4.24 +/- 2.62 ng/mL) when stimulated with 150 ng/mL GH than cultures of cells from normal stature subjects (12.3 +/- 2.5 ng/mL; 0.005 less than P less than 0.01). The cultures compared had a similar cell density. A similar significant difference in IGF-I secretion occurred between cultures of pygmy and control cells stimulated with 10 ng/mL GH. These data are consistent with previous in vivo studies in which pygmies failed to increase IGF-I and exhibit metabolic responses to exogenous GH. PMID- 2551918 TI - Calmodulin quantification and immunolocalization in developing embryonic orofacial tissue. AB - Temporally and quantitatively coordinated synthesis of cyclic adenosine monophosphate appears to be critical for normal development of the mammalian secondary palate. Calmodulin has been implicated as being involved in mediating the activity of a number of fundamental calcium-regulated intracellular enzyme systems including phosphodiesterases, adenylate cyclase, and a variety of kinases, all of which may regulate or be regulated by intracellular cAMP. Calmodulin levels were thus quantified, and endogenous calmodulin was immunolocalized in developing palatal tissue in vivo and in embryonic palatal mesenchymal cells in vitro. Endogenous palatal calmodulin levels, determined by radioimmunoassay, showed little variation during the period of murine palatal ontogenesis and averaged 0.23 ng/micrograms protein. Murine palate mesenchymal cells in monolayer, either in logarithmic growth or at confluency, contained similar levels of calmodulin. In palate mesenchymal cells in primary culture, specific anti-calmodulin staining was confined to the cell cytoplasm and was concentrated in the perinuclear region. Since immunostaining for calmodulin appeared to be associated with discrete cytoplasmic filaments, distribution of actin and tubulin were investigated. Immunostaining for tubulin in these cells was also localized to the perinuclear region, while immunolocalization of actin showed staining patterns, reflective of stress fibers, which were quite different from those seen for calmodulin. Immunostaining was seen in vivo in all regions of the palatal epithelium with superficial peridermal cells staining most intensely. Specific immunostaining was also evident in palatal mesenchyme, where a pericellular distribution was seen. Staining patterns were similar throughout the period of palatal ontogeny. In addition, a sharply defined localization of calmodulin to cartilagenous extracellular matrix was noted. This study provides a useful initial approach toward understanding the role calmodulin may play in embryonic orofacial development. PMID- 2551919 TI - Bradykinin potentiates LH-induced follicular rupture in the rat ovary perfused in vitro. AB - The possible role of bradykinin as a modulator of LH-induced ovulation was investigated using a model of the in-vitro perfused rat ovary. Ovaries from immature rats, primed with pregnant mare's serum gonadotrophin (PMSG; 20 IU), were perfused in vitro for 20 h, starting on the morning of induced proestrus. Stimulation in vitro with luteinizing hormone (LH; 0.1 microgram/ml) resulted in 3.4 +/- 1.2 ovulations per treated ovary, whereas no ovulations occurred in the unstimulated group. Bradykinin (5 microM) added to the perfusion system at 0, 2.5, 5, 7.5 and 10 h gave two ovulations in one ovulating ovary out of five ovaries perfused. When LH was combined with bradykinin, added to concentrations of 1 microM and 5 microM at the above-mentioned five time points, the numbers of ovulations were 12.2 +/- 2.7 and 15.6 +/- 3.7 per treated ovary, respectively. Bradykinin (5 microM), administered as a single dose concomitantly with LH, resulted in no further increase in the ovulation rate (3.6 +/- 1.6). Bradykinin did not affect cyclic AMP or steroid release from unstimulated or LH-stimulated ovaries. These data indicate a role of bradykinin in the ovulatory process of the rat, potentiating LH-induced ovulations. PMID- 2551920 TI - In-vivo transperitoneal fertilization. AB - We present the technique of in-vivo transperitoneal fertilization (IVTPF) as a first approach to infertility treatment in couples with male subfertility or unexplained factors. The technique is statistically less successful but also less invasive than either gamete intra-Fallopian transfer (GIFT) or in-vitro fertilization - embryo transfer (IVFET) and offers considerable advantages over intrauterine insemination (IUI). The IVTPF technique involves transperitoneal transfer of processed spermatozoa within the pouch of Douglas after induction of ovulation. We report our 4-year experience with IVTPF which includes 136 treatment cycles in 89 couples. Eight pregnancies were achieved in 89 patients (9%) and 136 treatment cycles (7%). Fifty-one patients (57%) received IVTPF for only one treatment cycle; seven of the eight IVTPF pregnancies occurred in this group. An ectopic pregnancy resulted in one of the eight IVTPF pregnancies (13%). The functional quality of the sperm in those couples who achieved pregnancy was statistically superior to those couples who did not conceive. However, pregnancy was also obtained in case of severe oligozoospermia. Based on our experience, we feel IVTPF to be a very reasonable first approach in patients with no pelvic pathology and with infertility secondary to male factors or unexplained causes. PMID- 2551921 TI - Production of human monoclonal anti-basement membrane zone (BMZ) antibodies from a patient with bullous pemphigoid (BP) by Epstein-Barr virus transformation. Analyses of the heterogeneity of anti-BMZ antibodies in BP sera using them. AB - We established three lymphoblastoid cell lines from a bullous pemphigoid (BP) patient's peripheral blood by means of EBV transformation, which produced human monoclonal anti-basement membrane zone (BMZ) IgG antibodies. A blocking immunofluorescence test using these MAbs, designated 5A, 5E, and 10D, revealed that 5A and 5E recognized the same or a closely associated epitope, but the epitope for 10D was completely different, 18 of 30 BP sera blocked the reactivity of 10D MAb and 17 sera blocked 5E, while 9 sera did not block the staining of either antibody. Immunoblot analysis demonstrated that both 5A and 5E MAbs reacted exclusively with a protein band of approximately 230 kD in normal human epidermal extracts. However, 10D did not show any protein band. 22 of 30 BP sera strongly reacted with the same 230-kD protein, while none of control sera showed such reactivity. These results clearly demonstrated the heterogeneity of anti-BMZ antibodies in terms of epitopes. These MAbs should be useful in future investigations concerning not only the immunopathology but also the biochemial and molecular analyses of the BP antigen. PMID- 2551922 TI - Lipocytes from normal rat liver release a neutral metalloproteinase that degrades basement membrane (type IV) collagen. AB - We report a proteinase that degrades basement-membrane (type IV) collagen and is produced by the liver. Its cellular source is lipocytes (fat-storing or Ito cells). Lipocytes were isolated from normal rat liver and established in primary culture. The cells synthesize and secrete a neutral proteinase, which by gelatin substrate gel electrophoresis and gel filtration chromatography, has a molecular mass of 65,000 D. The enzyme is secreted in latent form and is activated by p aminophenylmercuric acetate but not by trypsin. Enzyme activity in the presence of EDTA is restored selectively by zinc and is unaffected by serine-protease inhibitors. In assays with radiolabeled soluble substrates, it degrades native type IV (basement membrane) collagen but not interstitial collagen types I or V and exhibits no activity against laminin or casein. At temperatures causing partial denaturation of soluble collagen in vitro, it rapidly degrades types I and V. Thus, it is both a type IV collagenase and gelatinase. The enzyme may play a role in initiating breakdown of the subendothelial matrix in the Disse space as well as augmenting the effects of collagenases that attack native interstitial collagen. PMID- 2551923 TI - Immune response to B19 parvovirus and an antibody defect in persistent viral infection. AB - B19 parvovirus has been shown to persist in some immunocompromised patients, and treatment with specific antibodies can lead to decreased quantities of circulating virus and hematologic improvement. A defective immune response to B19 parvovirus in these patients was shown by comparison of results using a capture RIA and immunoblotting. In normal individuals, examination of paired sera showed that the dominant humoral immune response during early convalescence was to the virus major capsid protein (58 kD) and during late convalescence to the minor capsid species (83 kD). In patients with persistent parvovirus infection, variable titers against intact particles were detected by RIA, but the sera from these patients had minimal or no IgG to capsid proteins determined by Western analysis. Competition experiments suggested that this discrepancy was not explicable on the basis of immune complex formation alone and that these patients may have a qualitative abnormality in antibody binding to virus. In neutralization experiments, in which erythroid colony formation in vitro was used as an assay of parvovirus activity, sera from patients with poor reactivity on immunoblotting were also inadequate in inhibiting viral infectivity. A cellular response to purified B19 parvovirus could not be demonstrated using proliferation assays and PBMC from individuals with serologic evidence of exposure to virus. These results suggest that production of neutralizing antibody to capsid protein plays a major role in limiting parvovirus infection in man. PMID- 2551925 TI - Decreased adrenergic neuronal uptake activity in experimental right heart failure. A chamber-specific contributor to beta-adrenoceptor downregulation. AB - The reduction of myocardial beta-adrenoceptor density in congestive heart failure has been thought to be caused by agonist-induced homologous desensitization. However, recent evidence suggests that excessive adrenergic stimulation may not produce myocardial beta-receptor downregulation unless there is an additional defect in the local norepinephrine (NE) uptake mechanism. To investigate the association between beta-adrenoceptor regulation and NE uptake activity, we carried out studies in 30 dogs with right heart failure (RHF) produced by tricuspid avulsion and progressive pulmonary artery constriction and 23 sham operated control dogs. We determined NE uptake activity by measuring accumulation of [3H]NE in tissue slices, NE uptake-1 carrier density by [3H]mazindol binding and beta-adrenoceptor density by [3H]dihydroalprenolol binding. Compared with sham-operated dogs, RHF dogs showed a 26% decrease in beta-adrenoceptor density, a 51% reduction in NE uptake activity, and a 57% decrease in NE uptake-1 carrier density in their right ventricles. In addition, right ventricle beta-receptor density correlated significantly with NE uptake activity and NE uptake-1 carrier density. In contrast, neither NE uptake activity nor beta-receptor density in the left ventricle and renal cortex was affected by RHF. Thus, the failing myocardium is associated with an organ- and chamber-specific subnormal neuronal NE uptake. This chamber-specific loss of NE uptake-1 carrier could effectively reduce local NE clearance, and represent a local factor that predisposes the failing ventricle to beta-adrenoceptor downregulation. PMID- 2551926 TI - Ischemia-induced loss of epithelial polarity. Role of the tight junction. AB - In proximal tubular cells ischemia is known to result in the redistribution of apical and basolateral domain-specific lipids and proteins into the alternate surface membrane domain. Since tight junctions are required for the maintenance of surface membrane polarity, the effect of ischemia on tight junction functional integrity was investigated. In vivo microperfusion of early loops of proximal tubules with ruthenium red (0.2%) in glutaraldehyde (2%) was used to gain selective access to and outline the apical surface membrane. Under control situations ruthenium red penetrated less than 10% of the tight junctions. After 5, 15, and 30 min of ischemia, however, there was a successive stepwise increase in tight junction penetration by ruthenium red to 29, 50, and 62%, respectively. This was associated with the rapid duration-dependent redistribution of basolateral membrane domain-specific lipids and NaK-ATPase into the apical membrane domain. Taken together, these data indicate that during ischemia proximal tubule tight junctions open, which in turn leads to the lateral intramembranous diffusion of membrane components into the alternate surface membrane domain. PMID- 2551924 TI - Molecular mimicry and myasthenia gravis. An autoantigenic site of the acetylcholine receptor alpha-subunit that has biologic activity and reacts immunochemically with herpes simplex virus. AB - The large majority of patients with the autoimmune disease myasthenia gravis characteristically have detectable antibodies against the acetylcholine receptor (AChR). We used synthetic peptides to identify antibodies in sera of myasthenia gravis patients reactive with the human acetylcholine receptor (HuAChR) alpha subunit, residues 160-167. Affinity purification of these antibodies, using the HuAChR alpha-subunit 157-170 peptide immobilized on thiopropyl-Sepharose, yielded IgG antibodies that bound to the native AChR and inhibited the binding of alpha bungarotoxin to the receptor. The HuAChR alpha-subunit 160-167 peptide demonstrated specific immunological cross-reactivity with a shared homologous domain on herpes simplex virus glycoprotein D, residues 286-293, by both binding and inhibition studies. Thus, HuAChR alpha-subunit, residues 160-167, elicits antibodies in myasthenic patients that binds to the native AChR protein and is capable of eliciting a biologic effect. Immunologic cross-reactivity of this "self" epitope with herpes simplex virus suggest that this virus may be associated with the initiation of some cases of myasthenia. PMID- 2551927 TI - Erythropoietin receptors in polycythemia vera. AB - The role of erythropoietin (EP) in polycythemia vera (PV) is controversial, with some experiments suggesting that erythroid progenitors in PV are exquisitely sensitive to EP and EP dependent, and others suggesting that PV progenitors are EP independent. We have examined the characteristics of the EP receptor (EP-R) on erythroid colony-forming cells (ECFC) from patients with PV. In contrast to normal ECFC, which have two classes of EP-R, with 20% showing high affinity (Kd = 0.13 nM; range, 0.04-0.20 nM) and the remainder lower affinity (Kd = 0.37 nM; range, 0.28-0.57 nM), PV ECFC show a single class of 851 low affinity EP-R with Kd = 0.72 nM (range, 0.36-0.85 nM). ECFC from patients with secondary (EP driven) polycythemia or anemia show two classes of EP-R (Kd = 0.18 and 1.10 nM, respectively). Attempts to remove tightly bound EP from putative high affinity EP R in PV did not reveal any higher affinity receptors. Determination of molecular size by crosslinking showed two proteins of 90 and 100 kD similar to those seen with normal EP-R. These studies indicate the PV ECFC have EP-R that are structurally similar to normal EP-R but lack the higher binding affinity for EP. PMID- 2551928 TI - Sonographic detection of a nonpalpable regressed germ-cell tumor in an atrophic testis. PMID- 2551929 TI - Oestrogen receptors in mucinous carcinoma of the breast: an immunohistological study using paraffin wax sections. AB - An immunohistological method (Shintaku-Said method) for the demonstration of oestrogen receptors in routinely processed paraffin wax embedded tissue was applied to 19 cases of mucinous carcinoma of the breast. Seventeen (89%) tumours showed variable degrees of positivity and two were negative. In eight cases the receptors were also assayed biochemically using a dextran-coated charcoal method, and the results of the two methods showed good correlation. No difference in the distribution of positive and negative cases was noted between pure and mixed mucinous tumours, and in the latter group the pattern of staining of the mucinous elements was similar to that seen in the solid elements. It is concluded that the major advantage of this method is its ability to offer for study the distribution of the receptors in individual cells and specific histological structures. The results also indicate that most mucinous carcinomas of the breast are oestrogen receptor positive, irrespective of whether they are pure or mixed type. PMID- 2551930 TI - Expression of laminin in benign and malignant sclerosing lesions of extrahepatic bile ducts. AB - The distribution of laminin, a basement membrane glycoprotein, was studied by an immunohistochemical technique in six samples of benign sclerosing lesions of extrahepatic bile ducts and in 11 sclerosing cholangiocarcinomas. The expression of laminin showed that benign glandular structures were surrounded by a mainly intact basement membrane. In sclerosing cholangiocarcinomas laminin was irregularly distributed, and in large areas totally absent. The findings suggest that deposition of basement membrane material is defective. Sclerosing cholangitis may be extremely difficult to distinguish from well differentiated sclerosing cholangiocarcinoma. This sample was small, but the diseases studied were relatively rare. Staining for laminin might be of additional use to surgical pathologists in the differential diagnosis of benign sclerosing lesions and sclerosing cholangiocarcinomas. PMID- 2551931 TI - Detection of human papilloma viruses in paraffin wax sections with biotinylated synthetic oligonucleotide probes and immunogold staining. AB - Human papilloma virus was detected by in situ hybridisation in routinely processed paraffin wax sections using a synthetically produced oligonucleotide probe, end-labelled with biotin, and amplified with anti-biotin-immunogold silver staining (anti-biotin-IGSS). This system proved more sensitive than amplification with streptavidin-biotinylated alkaline phosphatase for detecting human papilloma virus type 16 in cervical tissues. The method was successfully combined with antigen staining for papilloma virus common antigens in skin and genital warts. This simple and quick method, using non-radioactively labelled synthetic probes, may be useful for the detection of other viruses in stored material and may be suitable for other double staining procedures. PMID- 2551932 TI - Dopaminergic activity of the antimuscarinic antiparkinsonian agents. AB - The antimuscarinic antiparkinsonian agents are widely used in psychiatric practice to reduce the extrapyramidal motor symptoms caused by the neuroleptic antipsychotic medications. Although the antimuscarinic antiparkinsonian agents are effective in reducing extrapyramidal symptoms, their use in conjunction with neuroleptic treatment of psychosis has been reported to antagonize the therapeutic effects of the neuroleptic; there are also several reports of the antimuscarinic antiparkinsonian agents variously causing psychotic syndromes, mood elevating and stimulant effects, stereotypy, dyskinesia, behavioral agitation, and drug dependence in both psychiatric and normal populations. These drug-related phenomena are generally attributed to the antimuscarinic properties of these agents. A large body of data, however, has shown that the antimuscarinic antiparkinsonian agents also function as potent, indirect dopamine-agonists. Benztropine, the most widely prescribed of these medications, is one of the most potent known inhibitors of presynaptic dopamine reuptake. These antiparkinsonian agents also have potent agonist activity at the noradrenergic synapse, as well as minor activity at the serotonergic synapse. This paper reviews neuropharmacologic evidence suggesting that significant neurophysiologic effects can result from the dopaminergic--and possibly noradrenergic--activity of the antimuscarinic antiparkinsonian agents, similar in some cases to those observed with amphetamine. Greater attention to these properties may aid in interpretation of clinical and research observations involving these so-called "antimuscarinic" agents. PMID- 2551933 TI - Autoradiographic distribution of thyrotropin-releasing hormone receptors in the African lungfish Protopterus annectens. AB - We used quantitative autoradiography to examine the distribution of thyrotropin releasing hormone (TRH) receptors in the central nervous system (CNS) of the African lungfish Protopterus annectens. We found that the distribution of TRH receptors throughout the CNS of the lungfish was heterogeneous with the highest concentrations (500-800 fmol/mg protein) in the olfactory bulb and telencephalon, moderately high concentrations (200-500 fmol/mg protein) in the diencephalon, and moderate (50-200 fmol/mg protein) to low (less than 50 fmol/mg protein) concentrations in the brainstem and spinal cord. Except for the motor nuclei of the cranial nerves and spinal cord, TRH receptors were concentrated in the acellular regions. In the telencephalon and diencephalon, the receptor density was inversely related to cellular density. These results provide a neuroanatomic and neuropharmacologic basis for further investigations of TRH in the African lungfish. PMID- 2551934 TI - Central acoustic tract in an echolocating bat: an extralemniscal auditory pathway to the thalamus. AB - To determine the sources and targets of auditory pathways that bypass the inferior colliculus in the mustache bat, we injected WGA-HRP in the medial geniculate body and related auditory nuclei of the thalamus as well as in the lower brainstem. We used electrophysiological methods to verify that the injection electrode was in an area responsive to sound. The only thalamic injections that produced retrograde transport to cells in auditory nuclei caudal to the inferior colliculus were those that included the suprageniculate nucleus. These injections labeled a group of large multipolar cells lying between the ventral nucleus of the lateral lemniscus and the superior olivary complex. Neurons in this cell group have also been shown to project to the deep layers of the superior colliculus in the mustache bat. The pathway revealed by these studies is almost identical to the "central acoustic tract" in which fibers course medial to the lateral lemniscus and bypass the inferior colliculus to reach the deep superior colliculus and the suprageniculate nucleus. PMID- 2551935 TI - Metabolic anatomy of brain: a comparison of regional capillary density, glucose metabolism, and enzyme activities. AB - Regional variations in capillary density, glucose utilization rate, and activities of the glycolytic enzyme lactate dehydrogenase and the mitochondrial enzyme cytochrome oxidase were compared in the rat brain. The distributions of capillaries and enzymes were studied by means of histochemical staining techniques, and glucose metabolism was measured by means of [14C]2-deoxyglucose autoradiography. Analysis of 18 gray and five white matter regions revealed a positive correlation between capillary density and glucose utilization rate. A negative correlation was found between capillary density and lactate dehydrogenase among gray matter structures. Analysis of capillaries and enzymes was also performed within laminated histological fields: hippocampus, olfactory bulb, and olfactory cortex. In general, this revealed reciprocal patterns of staining for lactate dehydrogenase and cytochrome oxidase. Capillary density paralleled cytochrome oxidase activity. The zones of intense staining for lactate dehydrogenase and cytochrome oxidase corresponded to the synaptic terminal fields of different input pathways. These findings demonstrate distinct distributions of a glycolytic and an oxidative enzyme within the brain which are at least partly associated with pathway specificity. PMID- 2551936 TI - Lesions of the basal forebrain alter stimulus-evoked metabolic activity in mouse somatosensory cortex. AB - The role that acetylcholine plays in processing sensory stimuli is beginning to be characterized; however, morphological correlates of cholinergic effects on activity patterns in sensory cortex are not available. To study this problem, unilateral neurotoxic lesions that depleted the necortex of acetylcholine were made in the basal forebrains of mice. The aim of these experiments was to study the effect of cholinergic depletion on stimulus-evoked activity in the barrel field of the mouse somatosensory cortex. One month following the lesion, 2 deoxyglucose (2DG) experiments were conducted on the lesioned and on normal mice while the animal received bilateral stimulation to the C3 whisker. The tissue was processed for acetylcholinesterase and cytochrome oxidase histochemistry and 2DG autoradiography. Evaluation of the column-like 2DG label evoked in the somatosensory cortex revealed that the activity on the lesioned side was significantly reduced in dimension and intensity from that in the normal hemisphere. On the normal side, the activated barrels averaged 641 microns in tangential width, were 76.5% above background in density, and extended from lamina I-V. On the lesioned side, the activated barrels were 485 microns in tangential width, 65.4% above background in density, and extended from lamina II V. In other cortical regions, outside the stimulus-evoked barrel field, 2DG activity values were similar on the normal and lesioned side. Additionally, both the pattern and intensity of the cytochrome oxidase staining within the barrel field displayed no differences in either hemisphere. These studies suggest that acetylcholine plays a significant role in the processing of somatosensory information. Neurotoxic lesions that diminish cortical cholinergic innervation cause a reduction of stimulus-evoked activity levels, while underlying metabolic activity is either not affected or recovers over time. PMID- 2551937 TI - Increased peripheral lymphocytes, lymphoid hepatitis and anaemia in African vervet monkeys seropositive to retroviruses. AB - Wild-caught African Vervet monkeys are commonly infected by Simian T-lymphotropic virus I (STLV1) and Simian immunodeficiency virus (SIV), yet the natural histories of these infections are largely unknown. Seropositivity was associated with increased total, T and atypical lymphocytes. In seropositive females there was mild, normocytic, normochromic anaemia. Lymphoid hepatitis was present in seven seropositive cases. African Vervets used in biomedical research, vaccine production and organ transplantation research are often infected by exogenous retroviruses which can be oncogenic and immunosuppressive in captive monkeys. Elimination of these infections may be possible by breeding Vervets in captivity. PMID- 2551938 TI - Histological changes in lungs of calves exposed to an aerosol of Pasteurella haemolytica. AB - An experiment was designed to study the interaction of Pasteurella haemolytica with an attenuated bovine herpesvirus 1 in calves. Low titre of the virus culture used for aerosol exposure failed to produce measurable interaction. However, the experiment provided the first opportunity to study the light-microscopic changes in lungs of calves (n = 3) to a low-dose exposure (5-min aerosol) of P. haemolytica A1 from a fresh 5-h log-phase culture. The histopathological study was confined to tissue exposed to only P. haemolytica. A limited macroscopic pneumonia was produced in ventral parts of cranial lobes. Four days after exposure, a typical reaction featured four zones. Zone 1a at the centre with acute inflammatory processes and necrosis of phagocytic cells was surrounded by a broad band of compacted, largely necrotic macrophages and polymorphonuclear leucocytes (PMNL) in alveoli of zone 1b. Necrosis was confined to zone 1. Zone 2a frequently occupied the remainder of the lobule with irregular distribution of congestion, oedema with a fibrinous component, and infiltration by numerous PMNL, macrophages and other mononuclear inflammatory cells. The narrow zone 2b was located between zones 1b and 2a and had oedema with a fibrinous component, numerous fibrocytes, few inflammatory cells and empty capillaries. It is suggested that zone 2 served to isolate zone 1 by surrounding it with nonfunctional tissue. The pathogenicity of P. haemolytica is discussed for uncompromised lungs and lungs compromised by virulent BHV1 infection. PMID- 2551939 TI - Detection of reactivating pseudorabies virus in tissue by immunoperoxidase technique. AB - Reactivation of pseudorabies virus (PRV) was induced in pigs by prednisolone treatment. The virus was re-isolated from nasal secretions of four, brain cortex of two and mandibular lymph node of three out of 12 pigs, respectively. The characteristic lesion of recurrently infected pigs was focal necrosis in the brain cortex and mandibular lymph node, accompanied by reactivating virus particles in the degenerating cells. Coincident with the pathological lesions, PRV antigen was detected in two of the brain cortex and six of the mandibular lymph node specimens by immunoperoxidase staining. These results suggest that the immunoperoxidase technique was more sensitive than virus isolation for demonstrating the reactivity of PRV in recurrently infected pigs. PMID- 2551940 TI - A case of tubular apocrine adenoma with syringocystadenoma papilliferum. AB - We report a new case of a Japanese man who developed a tubular apocrine adenoma in association with syringocystadenoma papilliferum. The differences in the histopathological and immunohistochemical findings in these 2 tumors are described. This is the first immunohistochemical study of tubular apocrine adenoma. PMID- 2551941 TI - Effects of sodium bicarbonate and sodium sesquicarbonate on animal performance, ruminal metabolism, and systemic acid-base status. AB - Six rumen-fistulated lactating Holstein cows were arranged in a replicated 3 x 3 latin square design with 3-wk periods and offered diets containing concentrate and corn silage in a 60:40 ratio (DM basis). Treatments were: 1) basal diet, 2) basal diet with 1% NaHCO3, and 3) basal diet with 1% sodium sesquicarbonate. There were no differences among treatments in milk production, milk protein, or 3.5% FCM, but sodium sesquicarbonate increased milk fat percentage (3.89, 3.94, 4.06%) compared with that of the control. Rumen pH was higher for cows fed buffered diets than for control cows. Urine pH was higher for cows fed NaHCO3 diet than for those fed sodium sesquicarbonate and control diets. No differences were detected among treatment means for molar percentage of isobutyrate, isovalerate, or total VFA, Dietary sesquicarbonate addition increased molar percentage of acetate, decreased propionate, and resulted in a higher acetate:propionate ratio compared with the cows fed NaHCO3. However, molar percentage of butyrate and valerate decreased in cows fed sodium sesquicarbonate when compared with those fed the control diet. No differences among treatment means were detected for blood pH, pCO2, or HCO3. PMID- 2551942 TI - Prolyl hydroxylase and procollagen III peptide levels in the sera from patients with collagen diseases and psoriasis. AB - Prolyl hydroxylase (PH) and procollagen III peptide (PIIIP) levels in 43 serum samples from various collagen diseases including 19 progressive systemic sclerosis (PSS) cases were simultaneously assayed and compared to those of 22 age matched psoriasis patients and 20 healthy controls in order to examine whether they can serve as useful parameters of fibrotic changes, especially in the skin. Serum PH levels were determined with enzyme immunoassay and serum PIIIP levels with radioimmunoassay. The average values of PH were 109% (n = 3, (p less than 0.001; generalized morphea, GM), 104% (n = 1; overlap syndrome, OS), 57% (n = 6; mixed connective tissue disease, MCTD), 48% (p less than 0.05; PSS), 33% (n = 5; dermatomyositis, DM), 28% (psoriasis), and 9% (n = 9; systemic lupus erythematosus, SLE), higher than the control levels, while those of PIIIP were 82% (OS), 57% (MCTD), 42% (p less than 0.01; PSS), 11% (GM), and 5% (DM) higher and 22% (psoriasis) and 28% (SLE), lower than the controls. These results indicate that both values were higher in the diseases with skin fibrosis and lower in those without it, suggesting that both assays generally reflect the state of actual progression of fibrosis at the point of determination. These two values had considerable correlation; however, GM, psoriasis and SLE patients had relatively higher values of PH than of PIIIP. This result suggests that the PH assay may also reflect the state of connective tissue activation; therefore the PIIIP assay may be more specific for pathological fibrotic process.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551943 TI - Cisplatin combination chemotherapy in squamous cell carcinoma and adenoid cystic carcinoma of the skin. AB - Seven patients with skin cancers, six with squamous cell carcinoma (SCC) and one with adenoid cystic carcinoma, were treated with cisplatin in combination with vindesine or adriamycin. Partial response was observed in three patients with squamous cell carcinomas: two cases with metastatic lung lesions and one with a primary skin lesion and lymph node metastasis. Two of the responding SCC had been resistant to previous chemotherapy, including peplomycin and mitomycin C. Multiple metastatic lesions of adenoid cystic carcinoma of the skin completely regressed after two courses of the combination chemotherapy with cisplatin and adriamycin. This report showed that cisplatin combination chemotherapy may be useful for the treatment of cutaneous squamous cell carcinoma, which is resistant to peplomycin, and adenoid cystic carcinoma of the skin. PMID- 2551944 TI - Paget's disease of the male breast--report of a case and histopathologic study. AB - We report a 61-year-old male with mammary Paget's disease. Physical examination revealed a slightly exudative erythema at the areola and a reddish, enlarged left nipple. No tumor or left axillary lymph nodes was palpable. He underwent a left modified radical mastectomy. Histologically, there was an intraductal carcinoma in the upper portion of the mammary ducts. The axillary lymph nodes that were examined were free of metastasis. Paget cells had neither estrogen nor progesterone receptors. We speculated that the histogenesis of Paget cells involved carcinoma cells that invaded the epidermis of the nipple. PMID- 2551945 TI - [Models for rotating spin labels and probes in proteins and membranes]. AB - A method of comparative analysis of ESR spectra has been proposed. It allows to distinguish between two models, slow reorientation of spin labels, and rapid rotation of the cone. Comparison of experimental data for a number of biological objects the theoretical predictions has shown that the rotation of nitroxyl fragment of spin labels can be described by the model of slow anisotropic rotation with correlation time 10(-6) less than or equal to tau less than or equal to 10(-8) s in conditions where the rotation of macromolecules is "frozen". PMID- 2551947 TI - Rate dependence of action potential duration and refractoriness in canine ventricular endocardium differs from that of epicardium: role of the transient outward current. AB - Previous studies have provided evidence for an important contribution of the transient outward current to the electrical activity of canine ventricular epicardium, but not endocardium. The present study examines the characteristics of action potential duration and refractoriness in these two tissue types. The time and rate dependence of changes in action potential duration and refractoriness observed in epicardium were significantly more accentuated than in endocardium. The restitution of action potential duration in epicardium paralleled the restitution of phase 1 amplitude of the action potential in this tissue. The correlation between phase 1 amplitude and action potential duration recorded from a large number of epicardial and endocardial preparations was significant under both steady state and restitution conditions. 4-Aminopyridine, a transient outward current blocker, decreased the time dependence of phase 1 amplitude and concomitantly decreased the time dependence of action potential duration in epicardium. 4-Aminopyridine abbreviated the action potential duration of epicardium at slow stimulation rates but had little effect or prolonged it at fast rates or after premature stimulation. (The availability of a transient outward current is relatively small after premature stimulation.) The data support the hypothesis that the prominent presence of a transient outward current in epicardium, but not endocardium, contributes to the differences in the time and rate dependence of action potential duration and refractoriness in the two tissue types. The results also demonstrate the effect of an outward current to prolong the action potential and the effect of an outward current blocker to abbreviate the action potential.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551946 TI - [3H]yohimbine binding to platelet alpha 2-adrenoceptors in depression. AB - [3H]Yohimbine binding to platelet alpha 2-adrenoceptors was studied in depressed patients and healthy volunteers. Where possible platelet binding measurement was repeated in depressed patients following treatment. Bmax of [3H]yohimbine binding did not differ significantly between depressed patients and control subjects and did not change with treatment in depressed patients. KD was significantly lower in female depressed patients, particularly in those who were post-menopausal. Multivariate analysis showed significant effects on KD of depression, season of testing and assay protein concentration. PMID- 2551948 TI - Intravenous immune globulin in patients with acquired immune deficiency syndrome. AB - Infection with the human immunodeficiency virus results in a progressive immune deficiency involving many components of the immune system. The major target cells for infection are CD4 antigen-bearing cells, which include predominantly, but not exclusively, the helper T cell subset and the monocyte-macrophage cell system. Hypergammaglobulinemia is a common and early feature of human immunodeficiency viral infection but is often associated with an inability to mount specific antibody responses to neoantigens and decreased in vitro B cell differentiation responses. On the basis of our data and reports of others, a major component of B cell hyperactivity is related to the direct activity of the human immunodeficiency virus and its proteins. We and others have documented that B cells of these patients lose the capacity to respond to both T cell-dependent and T cell-independent stimuli in vitro. Thus the human immunodeficiency virus can profoundly influence B cells, causing both "stimulation" (vis-a-vis hypergammaglobulinemia) and "suppression" (poor specific antibody responses). However, the memory cells are probably not suppressed, and their dysfunction may be directly related to the extent of helper cell dysfunction. This explanation lends itself well to the clinical observation that children (who have not developed memory to recall antigens) with human immunodeficiency viral infection have more profound defects in specific antibody responses than do adults (who have an existing pool of memory cells). Thus the main rationale for intravenous immune globulin in acquired immune deficiency syndrome is to provide antibody replacement.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551949 TI - Intravenous immune globulin prophylaxis in recipients of a marrow transplant. AB - Bone marrow transplantation is a worldwide activity involving more than 250 transplant centers in more than 40 countries. The ultimate success of the procedure depends in large measure on supportive care of the transplant recipient. A major component of this supportive care is the prevention of opportunistic infections. This article reviews the nature of immunoglobulin class and subclass deficiency after marrow transplantation and the role of intravenous immune globulin in preventing infections and associated complications. PMID- 2551950 TI - Effects of growth hormone-releasing hormone (GHRH) on densely granulated somatotroph adenomas and sparsely granulated somatotroph adenomas in vitro: a morphological and functional investigation. AB - The effects of growth hormone-releasing hormone (GHRH) were studied on densely granulated somatotroph adenoma cells and sparsely granulated somatotroph adenoma cells in culture by measuring release of growth hormone (GH) as well as ultrastructural morphometrical parameters and comparing them with those of control adenoma cells. Both types of adenoma cells cultured with GHRH showed similar increases of GH release into culture media and exhibited similar increases in cytoplasmic volume densities (CVD) of endoplasmic reticulum and Golgi apparatus and decreases in CVD of secretory granules and secretory granule diameter. These results indicate that (1) both types of somatotroph adenoma cells react similarly to GHRH stimulation, despite their morphologic differences, and (2) GHRH stimulates GH synthesis as well as GH release by somatotroph adenoma cells. PMID- 2551951 TI - Effects of the antileukemic drug L-asparaginase on sex hormone-binding globulin: studies in vivo and in vitro. AB - L-asparaginase, an antineoplastic drug used in the treatment of acute lymphoblastic leukemia (ALL), has been previously shown to inhibit the hepatic synthesis of thyroxine-binding globulin (TBG). In two children treated by this drug for ALL, a dramatic decrease in serum sex hormone-binding globulin (SHBG) concentrations was also observed. Serum SHBG levels were still below normal 10 days after L-asparaginase withdrawal. To ascertain whether this reduction was due to the inhibition of SHBG synthesis, SHBG was measured by an immunoradiometric assay (IRMA) in the medium from human hepatoblastoma-derived cells, Hep G2 cells, grown in the absence or presence of graded amounts of the drug from 0.1 nM to 0.1 mM. The results showed a dose-dependent inhibition of SHBG synthesis, with a 50% reduction of SHBG in the medium, assayed by IRMA, using 250 nM L-asparaginase. Furthermore, a time-dependent inhibition was observed using a fixed concentration of the drug (50 nM) added for variable time intervals (1-4 days). These data suggest that the changes observed in vivo are likely due to the inhibitory effect exerted by the drug on SHBG synthesis. This action is not specific, but is part of a general effect at the hepatic level. PMID- 2551952 TI - Childhood onset inclusion body myositis mimicking limb-girdle muscular dystrophy. AB - Inclusion body myositis was initially recognized in patients with "steroid resistant polymyositis" and subsequently in patients with other immune-mediated disorders. The finding of inclusion body myositis in a patient diagnosed for 30 years as having limb-girdle muscular dystrophy suggests yet another patient pool that may harbor this entity. PMID- 2551953 TI - Modulation of the responses of fetal sheep to adrenergic stimulation by adrenal demedullation and chemical sympathectomy. AB - The responses to sympathetic stimulation of fetal sheep adrenal-demedullated or sympathectomised by infusion of guanethidine sulphate have been studied. Sympathetic responses in such denervated or sympathectomised fetuses was studied by intravenous infusion of adrenaline or noradrenaline at about 0.4 micrograms/min per kg. This infusion increased plasma concentration 100-200 fold and there was no significant difference between the control fetuses and those in the vasrious treatment groups. Catecholamine infusions at these rates normally have little effect upon fetal blood gas and pH values, but in adrenal demedullated fetuses adrenaline infusion drepressed fetal arterial PO2 by 4-6 mmHg (P less than 0.05). The heart rate and blood pressure responses to catecholamine infusion in sympathectomised fetuses was, as expected, much increased. Similar observations were made on adrenal-demedullated fetuses, an unexpected finding, and this is taken to illustrate loss of the adrenal medulla is associated with enhanced responsiveness to adrenergic stimulation in peripheral tissues. The majority of the endocrine and metabolic responses, as reflected in fetal plasma concentrations of ACTH, cortisol, insulin, glucose, lactate and fatty acids, to catecholamine infusion were similarly much enhanced by adrenal-demedullation and chemical sympathectomy. Of particular note was a substantial increase in the responsiveness of the fetal adrenal, as reflected in plasma cortisol, to stimulation by ACTH, a change that usually induces labour, but not so in the present sheep. The results on increased sensitivity in adrenal demedullated fetuses are discussed in relation to likely tissue mechanisms mediating the changes. PMID- 2551954 TI - Clinical epidemiology of chronic constipation. AB - Using health statistics from the United States and England and Wales, we review the epidemiology of constipation and possible etiologies of this disorder as suggested by its epidemiologic distribution. The analysis revealed that constipation is one of the most common chronic digestive disorders in the United States, affecting one of every 50 people. The occurrence of constipation increased with advancing age, showing an exponential increase in prevalence after the age of 65. The age distribution of constipation was similar in both sexes, but overall constipation was three times more common in women than in men. Constipation more frequently affected nonwhites than whites, and people from families with low income or less formal education. The characteristic epidemiologic pattern of constipation suggests the influence of environmental factors. Insufficient dietary fiber is widely believed to be a major cause of constipation, yet it is difficult to devise a mechanism by which dietary fiber alone could produce the marked differences observed between gender, race, and socioeconomic status. Recent evidence suggests that disruption of neural regulation of colonic motility plays an important role in the development of chronic constipation. This loss of neural regulation may result from mechanical damage to the pelvic nerves due to childbirth or pelvic surgery, exposure to environmental toxins (e.g., organochlorine insecticides or heavy metals), or possibly exposure to an infectious agent. Other environmental factors that may play a role in the pathogenesis of chronic constipation have not yet been elucidated. Consequently, studies examining the epidemiology of chronic constipation are important for providing insight into potential environmental risk factors relevant to the etiology of this disorder. PMID- 2551955 TI - Gastric linitis plastica with metastases to the colon: a mimic of Crohn's disease. AB - A 60-year-old woman had progressive lower abdominal pain, nonbloody diarrhea, and weight loss, followed by severe epigastric pain and dysphagia. Radiographic evaluation of the colon showed segmental strictures which were interpreted as Crohn's disease. Medical treatment was not helpful. Neither gross endoscopic appearance nor multiple biopsies of the esophagus, stomach, and colon were diagnostic. Finally, laparotomy with full-thickness biopsies of the stomach and colon revealed linitis plastica. The clinician should be alert to colonic metastases from gastric linitis plastica, for it can produce focal or segmental strictures, mimicking more common colonic diseases such as Crohn's disease. A full-thickness biopsy is often necessary for a firm diagnosis. We review the literature on this occurrence, highlighting the clinical and radiologic spectrum, as well as the organ systems most often affected when gastric linitis plastica metastasizes. PMID- 2551956 TI - Small hepatocellular carcinoma presenting as intrabiliary pedunculated polyp and obstructive jaundice. AB - A rare autopsy case of small hepatocellular carcinoma presenting as an intrabiliary pedunculated mass is reported. The patient, 60-year-old man, presented with obstructive jaundice and died 1 month later. At autopsy, an intrabiliary pedunculated tumor measuring 3.5 x 2.0 x 2.0 cm was found in the hepatic, right hepatic, and right posterior segmental bile ducts. The intraductal tumor was attached to the segmental bile ductal wall, where a tumor measuring 2.0 x 0.9 cm was found in the parenchyma. The parenchymal tumor was continuous with the intrabiliary mass. Histologically, the tumor was a hepatocellular carcinoma showing acidophilic granular cytoplasm with occasional hyaline globules and was structurally arranged in a vague trabecular pattern. Immunohistochemically, tumor cells were positive for alpha-fetoprotein and alpha 1-antitrypsin, but was negative for keratin. We present a brief review of the literature of such icteric hepatocellular carcinomas. PMID- 2551957 TI - Fine needle aspiration with Menghini needle biopsy. PMID- 2551958 TI - Renin messenger RNA localization in congenital mesoblastic nephroma using in situ hybridization. AB - Renin messenger (m) RNA distribution was studied in congenital mesoblastic nephroma, a usually benign renal tumour of early infancy which may be associated with excess renin production and hypertension. Using in situ hybridization with synthetic radiolabelled oligonucleotide probes combined with immunohistochemical studies, renin expression was found in areas of tumours containing recognizable cortical structures including glomeruli and tubules. Renin mRNA was also detected in vessels and larger vascular spaces within the tumour not associated with cortical structures. Cells in the tumour vessel walls and sinusoids which expressed renin also stained positively for vascular smooth muscle-specific alpha actin. PMID- 2551959 TI - Adrenaline infusion in man increases muscle sympathetic nerve activity and noradrenaline overflow to plasma. AB - This study was conducted to determine if muscle sympathetic nerve activity (MSA) and/or the neuronal release of noradrenaline per impulse are modulated by adrenaline in the physiological plasma concentration range. We gave step-wise infusions of adrenaline (0.05-0.6 nmol/kg per min) to 10 healthy young men and measured: intra-arterial blood pressure; heart rate; central venous pressure (CVP); efferent MSA (microneurography in the peroneal nerve); arterial (brachial artery) and femoral venous plasma concentrations of noradrenaline, and the spillover of noradrenaline to arterial and venous plasma (radiotracer infusion). The infusion of adrenaline caused a fall in diastolic blood pressure and tachycardia, and was associated with increases in MSA and noradrenaline spillover. These observations suggest that part of the adrenaline-induced increase in transmitter release is due to enhanced nerve impulse activity, but such a conclusion rests on the absence of diffusion limitations from the site of noradrenaline infusion into the blood stream. After termination of adrenaline infusion the tachycardia and elevated plasma noradrenaline levels persisted, but these changes were probably due mainly to a profound increase in nerve activity. Concurrently, there was a reduction in CVP which may have triggered the increase in efferent sympathetic nerve activity. Infusions of adrenaline did not influence the clearance of noradrenaline from arterial plasma, but the fractional extraction over the leg was moderately reduced, indicating that more arterial noradrenaline is recovered in venous plasma during adrenaline infusion. The present data suggest that the reasons for the adrenaline-induced increase in noradrenaline release are complex, but they are consistent with the hypothesis that stress levels of adrenaline enhance sympathetic nerve activity, and that circulating adrenaline may modulate both haemodynamic and neural responses to stress. PMID- 2551960 TI - Properties and heterogeneity of human fetal pre-B cells transformed by EBV. AB - A series of 75 EBV-transformed pre-B and B-cell lines from fetal bone marrow at 14 to 18 wk of gestation was cloned for phenotypic, functional and molecular genetic studies. B-cell type volume regulation in response to hypotonic stress, low level CD9 (BA2), and, perhaps biased by our use of EBV, functional EBV receptors with expression of CD21 (B2) determinants characterized the most immature cells detected. These "B-progenitors" contained EBV genomes, maintained Ig H and L chain (as well as TCR) constant region genes in germ-line configuration and did not express other B, T, or myeloid lineage-associated surface markers including CD20 and MHC class II determinants. Such cells may represent B progenitor cells preceding classical pre-B-lymphocytes in pathways of B cell differentiation. Reminiscent of Abelson virus-induced transformation of immature murine B cells, EBV transformability together with the above properties may be the earliest markers of B lineage commitment in man. The expression of MHC class II Ag, CD20, C mu-H and then L chain rearrangements and expression followed in less immature pre-B lymphocytes and permitted a classification of lines into discrete subgroups of increasing maturity. The physical organization of the H chain locus in a given line was a stable characteristic. However, fetal pre-B cell lines showed considerable intraclonal heterogeneity with respect to H chain gene expression and that of differentiation markers such as CD20. Subcloning experiments indicated that this heterogeneity reflected clonally stable expression patterns distributed among subclones in an all-or-none fashion. The induction, by IL-6, of L chain expression in some but not all of these clones was linked to the presence of C mu transcription products, consistent with a possible regulatory role of mu protein in L chain rearrangement and expression. Although pre-B cell differentiation likely follows inherent programming, external signals seem able to hasten development along prescribed, hierarchical differentiation pathways. PMID- 2551961 TI - Modulation of colony-stimulating factor-1 receptors on macrophages by tumor necrosis factor. AB - The effect of murine rTNF-alpha on the binding of human 125I-rCSF-1 to murine thioglycolate-elicited peritoneal exudate macrophages (PEM) was investigated. At 4 degrees C, 125I-CSF-1 binding to PEM was inhibited by preincubation with human rCSF-1, but not by other cytokines. When PEM were incubated with various cytokines at 37 degrees C, murine rTNF-alpha caused greater than 90% decrease in 125I-CSF-1 binding. This decrease was time, temperature and TNF dose dependent, and was not affected by preincubation with cycloheximide. The reduction in CSF-1 binding activity was reversed by prolonged incubation at 37 degrees C even in the presence of TNF. However, PEM preincubated with TNF subsequently washing free of residual TNF resulted in a rapid recovery of CSF-1 binding. This recovery of CSF 1-binding activity required protein synthesis. Binding studies suggested that the decrease in 125I-CSF-1 binding was most likely caused by a reduction in the number of CSF-1 receptors. In addition, preincubation with TNF at 37 degrees C inhibited 125I-CSF-1 binding on mononuclear phagocytes, including the macrophage cell line J774, bone marrow-derived macrophages, and nonelicited macrophages from three different strains of mice. In contrast, 125I-murine rTNF-alpha binding to PEM was not inhibited by preincubation with CSF-1 at 4 degrees C or 37 degrees C. These data suggest that TNF may play a role in the modulation of receptor expression on blood cells, and may point to a role for this pleiotropic cytokine in the regulation of hemopoiesis. PMID- 2551962 TI - Staphylococcal enterotoxin B and toxic shock syndrome toxin-1 bind to distinct sites on HLA-DR and HLA-DQ molecules. AB - Staphylococcal enterotoxins (SE) activate human T cells in vitro. This requires the presence of Ia+ accessory cells but does not require processing of the toxin by the accessory cell. We and others have recently demonstrated direct binding of SE to human MHC class II molecules. In this study, we have compared in detail the ability of class II molecules to bind two SE, toxic shock syndrome toxin-1 (TSST 1) and SEB. Scatchard analysis of equilibrium binding data indicate that SEB binds to Ia+ human cell lines with a 10-fold lower affinity than TSST-1. Likewise, SEB precipitates HLA-DR alpha- and beta-chains from detergent lysates of Ia+ cells less efficiently than TSST-1. The binding of TSST-1 and SEB to transfected L cells expressing HLA-DR and HLA-DQ gene products was differentially inhibited by anti-HLA-DR mAb. There was virtually no cross-inhibition of TSST-1 and SEB in competitive binding assays. We conclude that TSST-1 and SEB bind to two MHC class II sites which can be distinguished by their relative accessibility to blocking by anti-class II mAb and heterologous toxin. PMID- 2551963 TI - Bactericidal defect of neutrophils in a guinea pig model of thermal injury is related to elevation of intracellular cyclic-3',5'-adenosine monophosphate. AB - PG of the E series inhibit major effector functions of polymorphonuclear leukocytes (PMN) by elevating intracellular cAMP. The present study investigated the involvement of this mechanism in the bactericidal defect of PMN induced by thermal injury in a guinea pig model. Peripheral and peritoneal exudate PMN harvested from thermally injured guinea pigs at 1 or 2 days postburn had decreased bactericidal activity against Pseudomonas aeruginosa and a marked increase in cAMP content. Production of PGE1 by these cells in the absence of exogenous PMN activators was also increased. Treatment of PMN in vitro or in vivo with nonsteroidal anti-inflammatory drugs (indomethacin, ibuprofen, and piroxicam) restored bactericidal activity to normal and concomitantly reduced cAMP content and PGE1 production. A concomitant reduction in cAMP content and PGE1 production was also observed as bactericidal activity of PMN returned to normal under natural conditions during 4 to 7 days postburn. The enhancement of PMN bactericidal activity mediated by NSAID was fully counteracted by purified PGE1, theophylline, and by cAMP itself. These results suggest that the bactericidal defect of PMN induced by thermal injury is related to elevation of cAMP and that PGE1 plays a significant role in this phenomenon. PMID- 2551964 TI - Studies of protein kinase C in the rat basophilic leukemia (RBL-2H3) cell reveal that antigen-induced signals are not mimicked by the actions of phorbol myristate acetate and Ca2+ ionophore. AB - Exogenous activators of protein kinase C such as PMA in combination with a Ca2+ ionophore (A23187), cause secretion in rat basophilic (RBL-2H3) cells,but they do so through stimulatory signals that are not the same as those generated by Ag or oligomers of IgE. On the one hand, the synergy between PMA and A23187 and the suppression of Ag-mediated signals (hydrolysis of inositol phospholipids and rise in concentration of cytosolic Ca2+) by PMA were totally dependent on protein kinase C. The loss of synergistic and inhibitory actions of PMA, for example, correlated with the loss of protein kinase C (as determined by immunoblotting techniques) when cells were continuously exposed to PMA. Furthermore, the permeabilization of RBL-2H3 cells resulted in the loss of both protein kinase C and the inhibitory action of PMA, but both were retained if cells were exposed to PMA before permeabilization Ag-induced secretion, on the other hand, was not as dependent on the presence of protein kinase C. The potent inhibitor of this enzyme, staurosporine, which blocked completely the secretory response to the combination of PMA and A23187, did not inhibit Ag-induced secretion except at concentrations (greater than 10 nM) that inhibited Ag-stimulated hydrolysis of inositol phospholipids as well. Also RBL-2H3 cells still showed some secretory response (approximately 25% of normal) to Ag when cells were depleted (greater than 98%) of protein kinase C by prolonged treatment with PMA. Previous studies have indicated that the secretory response to PMA and A23187 is much lower than that elicited by Ag when the concentrations of stimulants were matched to give the same increase in concentrations of cytosolic Ca2+. PMID- 2551965 TI - Endogenous receptor-bound urokinase mediates tissue invasion of human monocytes. AB - Macrophages have a marked capacity to invade tissue in the course of cellular immune reactions that is thought to be based on the action of urokinase (u-PA). u PA is an ubiquitous serine protease that converts the zymogen plasminogen into the active protease plasmin. u-PA binds to specific receptors on the macrophage thereby enabling the cell to degrade interstitial tissue in the microenvironment. Two cytokines produced in the course of cellular immune reactions, IFN-gamma and TNF-alpha, increase the number of u-PA receptors on human cultured monocytes from 14,000 to 64,000 and 30,000 receptors/cell, respectively. We used an amnion invasion assay to investigate whether activated human monocytes exhibit an enhanced capacity to invade interstitial tissue in correlation to the increased numbers of u-PA receptors. We show in this study that IFN-gamma, which increases the number of endogenously occupied and saturable u-PA receptors, causes a threefold increase of monocyte invasion into amnion tissue in comparison to control cells. The anti-u-PA mAb MPW5UK, which blocks the activity of u-PA, inhibits monocyte invasiveness significantly. In contrast, TNF-alpha, which increases only the number of saturable u-PA receptors on monocytes, does not enhance their invasiveness. This finding suggests that only endogenously occupied u-PA receptors are instrumental in monocyte invasiveness. This conclusion is further supported by the findings that: 1) saturation of monocytes with u-PA does not further increase their invasiveness and that 2) plasminogen-activator inhibitor-2, a specific inhibitor of u-PA associated with endogenously occupied, but not of u-PA bound to saturable receptors, inhibits monocyte invasiveness completely. PMID- 2551967 TI - Specific detection of Haemophilus influenzae type b lipooligosaccharide by a polymyxin B monoclonal antibody assay. AB - A monoclonal antibody (MAb)-based enzyme immunoassay was developed for detection of Haemophilus influenzae type b (Hib) lipooligosaccharides (LOS). The high affinity of polymyxin B for lipid A was used to bind the Hib LOS to microtiter wells. The immobilized LOS was detected with MAbs directed against the oligosaccharide component of Hib endotoxin. Hib LOS concentrations were measured in in vitro samples and in cerebrospinal fluid (CSF) sample obtained from rabbits with experimental Hib meningitis. The sensitivity of the assay was 1 ng LOS/ml sample and the results obtained with this assay correlated significantly with those obtained with the standard Limulus amebocyte lysate assay. This new assay provides a method for specific detection of Hib LOS in CSF samples and in aqueous laboratory fluids. This general methodology should also be useful for experimental research involving specific LPS/LOS molecules. PMID- 2551966 TI - Transient activation of the NADPH oxidase through Fc gamma RI. Oxidase deactivation precedes internalization of cross-linked receptors. AB - It is well known that Fc gamma R mediate the rapid release of agents of inflammation and, in addition, play an important role in the uptake of stimulatory antibody complexes. Activation of the FcR for human IgG1 (Fc gamma RI) on human monocytic cells triggers a transient activation of the NADPH oxidase. In this study, we tested the possibility that transience of the NADPH oxidase activation might have been the result of rapid internalization of cross linked Fc gamma RI. Stimulatory receptor moieties were formed by cross-linking Fc gamma RI with receptor-specific mAb that are known to trigger superoxide anion release. The formation of the stimulatory receptor units was determined by quantitating the rate of superoxide anion production through its reduction of cytochrome c. This rate has been found to correlate with the rate of binding of cross-linking antibody and, therefore, the rate of formation of the stimulatory moieties (receptor aggregates). Internalization of cross-linked Fc gamma RI was measured by quantitation of cell-associated FITC-labeled Fc gamma RI-specific mAb resistant to acid elution. We found that cross-linking antibody bound to Fc gamma RI continued to be taken up by the cells well after cessation of oxidase activity. The constant rate of uptake and the differential effect of temperature on these two functions suggested that they are separately regulated. Quantitation of cross-linked receptors that were inactive, i.e., no longer stimulating superoxide anion production, indicated that 50% of internalizable, and therefore cross-linked, Fc gamma RI remained on the surface after oxidase activity had ceased. This evidence of cessation of oxidase activity before the endocytic uptake of mAb/R stimulatory units indicates that the activated state of surface cross-linked Fc gamma RI is of brief duration and that occupation of the receptors by cross-linking-ligand does not sustain the activated state of the receptor. Thus, Fc gamma RI-mediated oxidase activation is temporally limited to the formation of the stimulatory receptor moiety. PMID- 2551968 TI - Comparison of three different activation methods for coupling antibodies to magnetisable cellulose particles. AB - The periodate and 1,1'-carbonyldiimidazole activation methods were compared with the cyanogen bromide procedure for coupling antibodies to magnetisable cellulose/iron oxide solid-phase particles. Fluoroimmunoassays for quinine, primaquine, normetanephrine and cannabinoids were employed to assess the binding properties of such coupled solid phases. The cyanogen bromide and 1,1' carbonyldiimidazole methods gave similar products in most cases, while the specific binding capacity of periodate coupled particles was between two and five times lower. Nevertheless, comparable standard curves could be obtained with solid phase coupled by each method. The periodate and 1,1'-carbonyldiimidazole methods are acceptable alternatives, notably for laboratories lacking the facility to handle the toxic cyanogen bromide. PMID- 2551970 TI - Southern blot analysis of clonal rearrangements of T-cell receptor gene in plaque lesion of mycosis fungoides. AB - T-cell populations of 22 plaque lesions from seven mycosis fungoides patients were studied for clonal rearrangement of the beta chain of the T-cell receptor (T beta) gene. All plaque lesions employed in this study showed clinically similar appearance. Histologically, all the biopsy specimens showed epidermotropism and the dermal infiltration of mononuclear cells including atypical cells. Histochemically, the majority of the infiltrated cells had surface markers of helper T cells. DNA extracted from skin lesions, peripheral lymphocytes, and lymph nodes revealed that the monoclonal expansion of T cells was different among patients and lesions. DNA extracted from the two skin lesions of case 1 revealed a clonal expansion of T cells. The rearranged bands persisted for about 1 year. In contrast, all lesions from cases 3-7 showed no rearranged band. Interestingly, three lesions from case 2 showed mixed type results, i.e., the monoclonal expansion of T cells was detected in one lesion but not in the other two lesions. Time course study of case 2 revealed that the same rearranged band became detectable in all three skin lesions and a lymph node about 1 year later. These results suggest that in plaque lesions of mycosis fungoides, there are various stages of detectable monoclonality of infiltrating cells, although the clinical appearance of the plaques was similar, and that the variety of monoclonality may reflect the long clinical course of this disease. PMID- 2551971 TI - Free radical reduction mechanisms in mouse epidermis skin homogenates. AB - Scavenging mechanisms for persistent free radicals were investigated using nitroxide-type radicals as model compounds. The free radical reducing activity of a) isolated thioredoxin reductase, a flavin containing oxidoreductase, b) skin homogenates, and c) the epidermis of hairless mice was studied by electron spin resonance spectroscopy. In all three systems, reduction rates of different classes of nitroxide free radicals exhibited the following order: oxazolidinoxy greater than piperidinoxy greater than dihydropyrroloxy. The main reductant for piperidinoxy radicals in mouse skin homogenate is ascorbic acid. Other reducing activities were stimulated by NAD(P)H and could be inhibited by N-ethyl maleimide, suggesting involvement of thiol-dependent processes. Mammalian thioredoxin, a competitive inhibitor of nitroxide reduction by thioredoxin reductase, significantly stimulates nitroxide scavenging in skin homogenate. Thioredoxin reductase did not significantly participate in nitroxide reduction in skin homogenates. At the surface of mouse epidermis a cationic dihydropyrroloxy nitroxide, which was stable in the presence of mammalian thioredoxin reductase was readily reduced. The epidermal reduction was inhibited by zinc, N-ethyl maleimide, and by heat (70 degrees C, 5 min). At least for mouse epidermis, reduction of a variety of nitroxides is a complex phenomenon involving enzymatic and nonenzymatic mechanisms and cannot be used as a specific assay for an enzyme, e.g., thioredoxin reductase. The study indicates the epidermis contains an effective antioxidant system that scavenges ascorbate-sensitive piperidinoxy nitroxides as well as more reducing radicals exemplified by dihydropyrroloxy nitroxides. PMID- 2551969 TI - [Virilizing tumors of the adrenal glands. Apropos of 3 cases]. AB - Three cases of virilizing tumors of the adrenal gland are reported. Two were malignant and are briefly summarized, and the other was benign. A review of the literature concerning benign virilizing tumors of the adrenal is included. PMID- 2551972 TI - Changes of electron spin resonance membrane fluidity in hexadecane-induced hyperproliferative epidermis. AB - To study some of the biochemical and physical states of membranes associated with hyperproliferation, the effect of topical hexadecane on membrane fluidity in guinea pig epidermis was investigated by electron spin resonance using a 5 doxylstearic acid spin labeling agent. Guinea pig epidermal cells were separated into three regions of keratinocytes by Percoll density gradient centrifugation. Membrane fluidity and Na+, K+-ATPase activity were higher in hyperproliferating epidermal cells than in control. The free cholesterol content and the molar ratio of free cholesterol to phospholipid were found to decrease significantly. Also elevated levels of palmitic acid, stearic acid and omega-3 unsaturated fatty acid derived from phospholipid were observed. Normal differentiation of epidermis was found to be accompanied by a decrease in membrane fluidity, whereas a relatively high membrane fluidity was maintained in the hexadecane-induced hyperproliferation. PMID- 2551973 TI - Latent and replicating forms of Epstein-Barr virus DNA in lymphomas and lymphoproliferative diseases. AB - Epstein-Barr virus (EBV) is associated with lymphomas and lymphoproliferative diseases that occur mainly in immunocompromised patients, but the role EBV plays in their pathogenesis is unclear. The evidence linking EBV etiologically to these disorders includes the presence of EBV DNA and nuclear antigens in the lesions and serologic evidence that some patients with these lesions are experiencing primary or reactivated EBV infections. These syndromes may represent proliferation of cells latently infected with EBV, but the possibility of viral replication has not been rigorously studied. DNA extracted from biopsies of 35 lymphoproliferative diseases was probed with regions of the EBV genome capable of distinguishing circular, episomal DNA found in latency from linear, replicating EBV DNA. All samples contained restriction fragments characteristic of fused termini, indicative of circular, latent genomes. Thirteen samples contained additional restriction fragments diagnostic of linear EBV DNA. Therefore, replicating EBV DNA is found in approximately 40% of EBV-associated lymphoproliferative disorders. PMID- 2551974 TI - Genital human papillomavirus infection in Panama City prostitutes. AB - Little is known of the natural history of genital human papillomavirus (HPV) infections in women from high-risk populations. Samples were collected from 183 Panama City prostitutes and assessed for HPV (filter in situ DNA hybridization) and for sexually transmitted agents. The cohort was followed for 8 mo; 51% of subjects completed four monthly return visits and 16% were sampled eight times. The proportion of women found infected with HPV increased significantly with increasing numbers of consecutive samples tested; 38 (21%) of 183 women were positive after one visit and 46 (82%) of 56 who completed six visits were infected. The pattern of viral detection over time was not random, which implied that most prostitutes were persistently infected with genital HPVs and that either scattered foci of infection or periodic reactivation of latent virus occurred. Our findings suggest that multiple sampling is necessary to accurately estimate HPV infection rates and to define whether patterns of DNA expression are present. PMID- 2551975 TI - An outbreak of gastroenteritis associated with acute rotaviral infection in schoolchildren. AB - In April 1988 a large outbreak of group C rotavirus infection associated with acute gastroenteritis occurred among schoolchildren and their teachers simultaneously at seven elementary schools in Fukui city, Japan. Of 3,102, 675 (21.8%) became ill. Clinical symptoms were mild, predominantly abdominal pain and vomiting, with diarrhea reported in only 27.6%. The outbreak subsided within 2 d. No pathogenic bacteria were found in fecal specimens; the virus particles detected by electron microscopy were morphologically indistinguishable from conventional infantile rotavirus. Immune electron microscopy showed that these virions formed large aggregates with convalescent serum and with the reference serum specific to group C rotavirus. Polyacrylamide gel electrophoresis showed similar RNA patterns for virus from this outbreak and typical group C rotavirus. PMID- 2551977 TI - A postviral chronic fatigue syndrome: a round table. PMID- 2551976 TI - Ineffectiveness of recombinant interferon-beta serine nasal drops for prophylaxis of natural colds. AB - Two randomized, double-blind, placebo-controlled trials during early autumn of 1986 and 1987 evaluated the efficacy and tolerance of recombinant interferon-beta serine (rIFN-beta ser) nasal drops for prevention of natural rhinovirus colds. In 1986, 9 X 10(6) units of rIFN-beta ser (139 subjects) or placebo (157) were administered once daily except Sundays for 4 w. Rhinovirus colds occurred in 2.8% of rIFN-beta ser recipients and 6.0% of placebo recipients during the treatment period (52% reduction, P = .3). In 1987, 24 X 10(6) units of rIFN-beta ser (186) or placebo (197) were given daily for 25 consecutive days. Rhinovirus colds developed in 6.3% of rIFN-beta ser recipients and 5.3% of placebo recipients. In each study, illness frequency and number of days with subjective colds did not differ between the groups. Recipients of nasal drops of rIFN-beta ser at either dosage did not differ in tolerance from placebo recipients. The lack of both prophylactic efficacy and nasal toxicity are in contrast to prior observations with nasal sprays of rIFN-alpha 2b. PMID- 2551978 TI - Re-implantation of homologous, cultivated osteoblast-like cells for improvement of bone regeneration. An animal study. AB - The regeneration of osseous tissues after implantation of rat osteoblast-like cells cultivated in vitro was investigated. A cell line of rat osteoblast-like cells originating from explants of four inbred rats (Lewis) was established. The osteoblast-like cells were cultivated in vitro and re-implanted after 3 passages in cell culture into artificial femur defects of the same inbred Lewis rats. The cultivated osteoblast-like cells were embedded in bovine bone gelatine and implanted together with hydroxylapatite granulate (12 animals), pyrolized calf bone (12 animals) or without any additional material (12 animals). These bone replacement materials were used as a carrier for the osteoblast-like cells. After 10, 20, 40 and 80 days animals were killed and the treated femora were taken for histological examination and determination of bone growth rate. It was demonstrated that the cultivated osteoblast-like cells in this experiment had no measurable effect on quantitative bone growth, but caused a significant difference in the histological bone regeneration pattern. PMID- 2551979 TI - [Geographic localization of three variants of herpes simplex virus type 1 in Japan]. AB - We previously reported that a variant of herpes simplex virus type 1 (HSV-1) designated variant type A was isolated at a much higher frequency in Western Japan than in Eastern Japan. We examined the localization of two other variant types designated B and C. The proportion of the type B and type C variants among 553 fresh HSV-1 isolates from 10 areas in Japan were 5.4% and 1.3%, respectively. The isolation frequency of type B variant in Eastern Japan was 8.3%. It was as high as 20.5% in Iwate area. The frequency in Western Japan was only 3.0%. Type C variant showed no specific geographic localization. These results were discussed in terms of characteristic mode of HSV infection and the history of the Japanese nation. PMID- 2551981 TI - [A biochemical analysis of experimental chemonucleolysis]. AB - We have studied the mechanical properties of enzyme-injected canine intervertebral discs by quasi-static compression tests, dynamic vibration tests and histology at 2 weeks (short-period group) and within 6 months (long-period group) after enzyme injection. The findings of each group were compared with those of normal discs as a control. The results were as follows; The quasi-static tests: In the short-period group, normalized stiffness of the discs was reduced significantly and the normalized creep rate was increased. The intradiscal pressures produced by external loading in short-period discs were almost half those of the control. In the long-period group such values tended to recover partially. The dynamic tests: In the short-period group, both elasticity and viscosity were reduced, while in the long-period group elasticity recovered but viscosity did not. These findings suggest that the shock-absorbing ability of enzyme-injected discs does not recover. PMID- 2551980 TI - [The assessment of the safety of vacuum extraction deliveries under routine epidural block]. AB - To assess the safety of vacuum extraction (V.E.) deliveries under continuous lumbar epidural block (E.B.) with Bupivacain, the modified Krebs score in CTG, Apgar score, cord arterial blood acid-base balance, hypoxanthine, CPK, CPK-BB, Neuron specific enolase and c-AMP were examined. A total of 74 full term oxytocin induced labors were divided into three groups: A) 21 cases by V.E. under E.B., B) 34 cases under E.B. only, and C) 19 cases without V.E. or E.B. There was no difference in age, gestational weeks in the three groups. However, the incidence of primiparas was highest in group A. Though CTG showed a temporary low Krebs score in group A within 30 minutes after the initiation of E.B., it was found that there was no significant difference between the three groups 30 minutes before parturition. The apgar score, cord arterial blood pH, PO2 and B.E. were also found to be similar in all three groups. Among various kinds of substances in the cord blood, both CPK-BB and c-AMP showed a striking rise in group A, compared to groups B and C. Therefore, the use of V.E. under E.B. might be hazardous to the well-being of the newborn infant. PMID- 2551982 TI - [A case of hepatocellular carcinoma probably producing renin]. PMID- 2551983 TI - [Ki-1 lymphoma histologically mimicking Hodgkin's disease]. PMID- 2551984 TI - Heparin and low molecular weight heparin: is anti-factor Xa activity important? PMID- 2551985 TI - Inhibition of platelet-dependent thrombosis by low molecular weight heparin (CY222): comparison with standard heparin. AB - We have compared the relative antithrombotic and antihemostatic effects of the very low molecular weight heparin CY222 with standard unfractionated heparin (SH) in a baboon model of platelet-dependent thrombosis. Thrombus formation was induced by placement of a thrombogenic device in an exteriorized femoral arteriovenous shunt under conditions of intermediate-shear blood flow. The device consisted of a collagen-coated cannular segment positioned proximal to two regions of expanded diameter exhibiting disturbed flow and stasis. Thrombus formation was measured in real time by indium 111-labeled platelet imaging. The collagen-coated surface accumulated thrombi composed largely of platelets, and the regions of disturbed flow were morphologically rich in fibrin and red cells. SH and CY222 were administered by continuous infusion for 1 hour. Although both heparin preparations abolished thrombus formation in the low-shear fibrin-rich regions at plasma levels less than 0.5 anti-Xa U/ml, platelet deposition onto the collagen surface was not reduced by either SH or CY222 at that dosage. These findings were consistent with previously observed therapeutic benefits of this level of anti-Xa activity in venous, but not arterial, thrombosis. Platelet deposition on the collagen was reduced in a dose-dependent manner by both SH and CY222 administered at doses between 1 and 5 anti-Xa U/ml. It is important to note that although heparin preparations produced profound and equivalent antithrombotic effects for platelet-dependent thrombus formation at comparable levels of anti-Xa activity, SH prolonged both the coagulation time and the bleeding time substantially more than did CY222.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551986 TI - Isolation and characterization of lipoproteins produced by human hepatoma-derived cell lines other than HepG2. AB - A total of six established human hepatoma-derived cell lines, including Hep3B, NPLC/PRF/5 (NPLC), Tong/HCC, Hep 10, huH1, and huH2, were screened for their ability to accumulate significant quantities of lipoproteins in serum-free medium. Only two cell lines, Hep3B and NPLC, secreted quantitatively significant amounts of lipoproteins. In a 24-h period the accumulated mass of apolipoproteins (apo) A-I, A-II, B, and E and albumin for Hep3B cells was 1.96, 1.01, 1.96, 1.90, and 53.2 micrograms/mg cell protein per 24 h, respectively. NPLC cells secreted no detectable albumin but the 24-h accumulated mass for apolipoproteins A-I, A II, B, and E was 0.45, 0.05, 0.32, and 0.68 micrograms/mg cell protein per 24 h, respectively. Twenty four-hour serum-free medium of Hep3B cells contained lipoproteins corresponding to the three major density classes of plasma; percent protein distribution among the lipoprotein classes was 4%, 41%, and 56% for very low density lipoprotein ("VLDL"), low density lipoprotein ("LDL"), and high density lipoprotein ("HDL"), respectively. NPLC was unusual since most of the lipoprotein mass was in the d 1.063-1.235 g/ml range. Hep3B "LDL", compared with plasma LDL, contained elevated triglyceride, phospholipid, and free cholesterol. Nondenaturing gradient gel electrophoresis revealed that Hep3B "LDL" possessed a major component at 25.5 nm and a minor one at 18.3 nm. Immunoblots showed that the former contained only apoB while the latter possessed only apoE. Like plasma VLDL, Hep3B "VLDL" particles (30.5 nm diameter) isolated from serum-free medium contained apoB, apoC, and apoE. "HDL" harvested from Hep3B and NPLC medium were enriched in phospholipid and free cholesterol and poor cholesteryl ester which is similar to the composition of HepG2 "HDL." "HDL" from Hep3B and NPLC culture medium on gradient gel electrophoresis had peaks at 7.5, 10, and 11.9 nm which were comparable to major components found in HepG2 cell medium. Hep3B cells, in addition, possessed a particle that banded at 8.2 nm which appeared to be an apoA II without apoA-I particle by Western blot analysis. The cell line also produced a subpopulation of larger-sized "HDL" not found in HepG2 medium. NPLC "HDL" had a distinct peak at 8.3 nm which by Western blot was an apoE-only particle. Electron microscopy revealed that "HDL" harvested from Hep3B and NPLC medium consisted of discoidal and small, spherical particles like those of HepG2. The "HDL" apolipoprotein content of each cell line was distinct from that of HepG2. ApoA-II at 35% of apolipoprotein distinguishes Hep3B "HDL" from HepG2, which contains only 10%.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2551987 TI - Active components of excitability in K+-depolarized ventricular muscle. AB - The steady-state and dynamic characteristics of excitability were assessed in isolated guinea pig papillary muscles depolarized with elevated [K+]o to resting potentials near -60 mV. Transmembrane potentials were recorded from fibers during application of low-amplitude current pulses used to analyze net changes in active membrane components of excitability in terms of elicited local responses and measure threshold current (Ith). Generated local responses were blocked entirely by tetrodotoxin and lidocaine, which increased steady-state Ith by more than 200%. In the absence of Na+ channel-blocking agents, local responses showed marked but characteristic attenuation in a time- and voltage-dependent manner by preceding subthreshold depolarizations, which concomitantly reduced excitability. However, local responses and excitability were also modulated by small changes in [Ca2+]o (+/- 0.7 mmol) and reduced by exposure to slow channel blockers and to Cs+. Thus these data suggest that while the Na+ channel is the primary active component of excitability in partially depolarized ventricular muscle, Ca2+ mediated and Cs+ -sensitive conductances may also participate, although to a lesser extent. These findings may help explain the frequency-dependence of excitability and conduction under conditions of ischemia in the intact heart. PMID- 2551988 TI - The interaction between indomethacin and captopril or enalapril in healthy volunteers. AB - In this study eight healthy volunteers were involved in a randomized, cross-over trial in which they were treated with either 25 mg of captopril b.i.d. or 20 mg of enalapril o.i.d. alone or in combination with 50 mg of indomethacin b.i.d. in order to detect a difference between both converting enzyme inhibitors when interacting with indomethacin. Before and after each 4-d treatment period, blood pressure (determined by random zero sphygmomanometer), body weight, plasma renin activity, angiotensin converting enzyme, plasma potassium, serum creatinine and the 24-h urinary excretion of 6-keto-prostaglandin F1 alpha were measured. Indomethacin attenuated the decrease of supine diastolic blood pressure during treatment with captopril, but not with enalapril. However, the initial decrease of blood pressure on captopril was greater than on enalapril. Both converting enzyme inhibitors had no effect on the urinary excretion of 6-keto-prostaglandin F1 alpha, while indomethacin reduced it. The results suggest a difference between captopril and enalapril in interaction with indomethacin. PMID- 2551989 TI - Plasma concentrations of calcitonin gene-related peptide increase during haemodialysis: relation to blood pressure. AB - The authors measured the plasma levels of calcitonin gene-related peptide (CGRP), the most potent vasodilatator known, during sequential ultrafiltration and haemodialysis in 12 patients using a radio-immunoassay method. Mean plasma levels of CGRP were 70.3 +/- 16.5 (mean +/- SE) pmol l-1 at the start of treatment, it increased to 85.3 +/- 17.6 pmol l-1 during ultrafiltration and to 114.5 +/- 25.3 pmol l-1 during dialysis. Systolic blood pressure decreased during haemodialysis. Plasma levels of CGRP were negatively correlated to systolic blood pressure before and at the end of dialysis, and changes in plasma levels of CGRP were strongly correlated to changes in systolic blood pressure. The increase in CGRP levels was not correlated to the fluid removal, toxin removal or changes in osmolality. The increase in plasma levels of CGRP observed during dialysis may be an important cause of dialysis induced vasodilatation and fall in blood pressure. PMID- 2551991 TI - Concentration of zinc and zinc-copper superoxide dismutase activity in red blood cells in normals and children with cancer. AB - Zinc concentrations and Cu, Zn superoxide dismutase activity in erythrocytes were investigated in 138 healthy children and in 35 children with cancer. The mean zinc concentration in the erythrocytes of healthy children was found to be age dependent. In the youngest group (children up to 1 year of age) the zinc concentration in erythrocytes is 18.8 +/- 3.4 micrograms/g Hb (5.89 +/- 1.23 mg/l packed cells), which is significantly lower than in other age groups. A strong logarithmic correlation (r = 0.327, p less than 0.0001 and r = 0.436, p less than 0.00001) was found between age and zinc concentration in erythrocytes, expressed as microgram/g Hb and as mg/l packed red cells, respectively. Cancer children were divided into two groups (neuro- and nephroblastoma). In the group of children with neuroblastoma no statistical differences in zinc concentration or enzyme activity were observed. In the patients with nephroblastoma, significantly higher zinc concentrations (p less than 0.05) were observed in erythrocytes. The changes of zinc concentration are accompanied by significant (p less than 0.02) decreases of enzyme activity. In this group of cancer children, statistically significant differences were observed in the zinc concentrations in erythrocytes (microgram/g Hb) between the second and the third stages of the disease. No correlation was observed between the concentration of zinc and enzyme activity in healthy children or in cancer children. PMID- 2551990 TI - Endothelial receptor-mediated binding of glucose-modified albumin is associated with increased monolayer permeability and modulation of cell surface coagulant properties. AB - Advanced glycosylation end products (AGE) of proteins accumulate in the vasculature with diabetes and aging, and are thought to be associated with vascular complications. This led us to examine the interaction of AGE-BSA as a prototype of this class of nonenzymatically glycosylated proteins subjected to further processing, with endothelium. Incubation of 125I-AGE-BSA with cultured bovine endothelium resulted in time-dependent, saturable binding that was half maximal at a concentration of approximately 100 nM. Although unlabeled normal BSA was not a competitor, unlabeled AGE-BSA was an effective competitor of 125I-AGE BSA-endothelial cell interaction. In addition, AGE modification of two alternative proteins, hemoglobin and ribonuclease, rendered them inhibitors of 125I-AGE-BSA binding to endothelium, although the native, unmodified forms of these proteins were not. At 37 degrees C, binding of 125I-AGE-BSA or gold-labeled AGE-BSA was followed by internalization and subsequent segregation either to a lysosomal compartment or to the endothelial-derived matrix after transcytosis. Exposure of endothelium to AGE-BSA led to perturbation of two important endothelial cell homeostatic properties, coagulant and barrier function. AGE-BSA downregulated the anticoagulant endothelial cofactor thrombomodulin, and induced synthesis and cell surface expression of the procoagulant cofactor tissue factor over the same range of concentrations that resulted in occupancy of cell surface AGE-BSA binding sites. In addition, AGE-BSA increased endothelial permeability, resulting in accelerated passage of an inert macromolecular tracer, [3H]inulin, across the monolayer. These results indicate that AGE derivatives of proteins, potentially important constituents of pathologic vascular tissue, bind to specific sites on the endothelial cell surface and modulate central endothelial cell functions. The interaction of AGE-modified proteins with endothelium may play an important role in the early stages of increased vascular permeability, as well as vessel wall-related abnormalities of the coagulation system, characteristic of diabetes and aging. PMID- 2551992 TI - Peritoneal tuberculosis, large ovarian thecoma, and an elevated serum CA 125 level mimicking ovarian cancer. AB - Laparotomy revealed peritoneal tuberculosis in a 32-year-old woman with pulmonary tuberculosis, ascites, a large pelvic mass, and an elevated serum CA 125 level. Postoperatively, after further treatment with antituberculosis agents, the CA 125 returned to normal and the patient has been well for 22 months. PMID- 2551993 TI - Free alpha subunit of human chorionic gonadotropin in women with non trophoblastic tumors. AB - Patterns of secretion of free alpha subunit in 242 women with non-trophoblastic tumors were studied. In 4 patients molecular forms of alpha subunit were investigated. In normal menstruating women, day to day fluctuations of alpha subunit (36 to 4871 ng/day) with a peak at midcycle were commonly observed. This makes measurement of the alpha subunit as a tumor marker difficult to interpret in this group of women. On the other hand, the secretion of alpha subunit in postmenopausal women was relatively stable (from 50 to 450 ng/day). Thus the incidence of elevated alpha subunit secretion (greater than 400 ng/day) in postmenopausal patients with non-trophoblastic tumors was as follows; cervical cancer (29.9%), ovarian cancer (50%), corpus cancer (37.5%), vulvar cancer (33.3%), lung cancer (53.3%), gastrointestinal tract cancer (80%). The elevation of the alpha subunit was unrelated to the histological type of cancer. Discordant secretion of hCG and its subunits were noted. The incidence of elevated hCG secretion (greater than 100 ng/day) for the same group of postmenopausal patients with non-trophoblastic tumors was 44.8% for cervical cancer, 100% for ovarian cancer, 62.5% for corpus cancer, 33.3% for vulvar cancer, 26.7% for lung cancer and 40% for gastrointestinal tract cancer. The incidence of isolated alpha subunit secretion in cervical cancer stages III and IV, lung cancer and gastrointestinal tract cancer (40-43%) was higher than that in other cancers (0 27%). For cervical cancer, the more advanced disease was associated with higher alpha subunit levels. The molecular forms of alpha subunit in 4 patients with non trophoblastic tumors were studied by gel HPLC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2551994 TI - Natural killer cell activity in hepatocellular carcinoma patients. AB - Twenty-four hepatocellular carcinoma (HCC) patients, 35 asymptomatic hepatitis B surface antigen (HBsAg) carriers and 30 normal healthy individuals, were included in this study to evaluate the natural killer (NK) cell population and NK cell activity, particularly before and after surgery for HCC patients. The NK cell population was stained with fluorescent labelled monoclonal antibody (leu 11b) and counted under fluorescent microscope. The NK cell activity was evaluated by Cr51 release assay and K562 cells line serving as target cells. The result indicated that there is no significant difference in NK cell population in HCC patients as compared with either the normal control or asymptomatic HBsAg carriers. (p greater than 0.05) However, significantly depressed NK cell activity was observed in asymptomatic HBsAg carriers and HCC patients as compared with that of the normal control. The NK cell activity of HCC patients was significantly depressed further after surgical removal of the tumor on the 1st postoperative week. Though the NK cell activity showed signs of recovery on the 2nd and the 4th postoperative week, it still did not return to the preoperative level even on the 4th postoperative week. PMID- 2551995 TI - Late onset hemorrhagic cystitis after allogeneic bone marrow transplantation. AB - Three patients with acute myeloblastic leukemia, thalassemia major and aplastic anemia experienced hemorrhagic cystitis on the 23rd, 35th and 36th day respectively after allogeneic bone marrow transplantation. The conditioning regimens before transplantation comprised cyclophosphamide with or without busulfan and total lymphoid irradiation. The hematuria lasted from 5 to 45 days and then subsided after treatment. Multiple factors including the toxicity of chemotherapeutic agents, viral infection and graft-versus-host reactions may contribute to late onset hemorrhagic cystitis after allogeneic bone marrow transplantation. Adequate treatment to minimize urothelial damage from chemotherapy, screening for viral infection and controlling graft-versus-host disease are mandatory in decreasing the complication of late-onset hemorrhagic cystitis after bone marrow transplantation. PMID- 2551996 TI - Lymphocyte transformation in patients with HBsAg-positive chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. AB - To evaluate cellular immunity, mitogen-induced lymphocyte transformation was assessed in 146 patients with chronic hepatitis B virus (HBV) infection, including 63 patients with chronic hepatitis (CH), 34 with liver cirrhosis (LC), and 49 patients combined with liver cirrhosis and hepatocellular carcinoma (HCC). The PHA-induced transformation indices (TI) were lower in patients with chronic HBV infection, especially in the LC and the HCC groups. The Con A-induced TI were also diminished in patients with chronic HBV infection. The mitogen-induced TI in patients with CH showed no significant difference in regards to different histological patterns, HBeAg/anti-HBe statuses, sex groups, age distributions, and SGPT levels. However, the mitogen-induced TI were substantially higher in CH patients with elevated serum AFP levels, especially in the group with AFP levels over 100 ng/ml. From these data, we conclude that: (1) impaired PHA-induced TI in patients with LC may partly explain the high risk factor of LC in the occurrence of HCC; (2) the reduction in TI induced by optimal concentration of Con A reflects the defective suppressor cell activity in patients with chronic HBV infection, even in patients with HCC; (3) no significant difference among the control group and patients with chronic HBV infection in PWM-induced TI implies that the response of B cells may be normal in patients with chronic HBV infection; (4) elevated mitogen-induced TI in CH patients with high AFP levels indicate that the activation of lymphocyte may contribute to the severity of inflammation and tissue damage. PMID- 2551997 TI - Malignant fibrous histiocytoma of the head and neck: report of 5 cases. AB - Malignant fibrous histiocytoma is a rare tumor in the head and neck region. This paper describes 5 cases, one was primary, and the remaining 4 cases were supposed to be secondary to previous radiotherapy for malignant diseases in the head and neck region. Three cases were in the maxillary sinus and two at the tongue base. All patients received surgical treatment with or without postoperative irradiation. Two patients survived more than 3 years after surgical intervention. PMID- 2551998 TI - Sodium channel gating in clonal pituitary cells. The inactivation step is not voltage dependent. AB - We have determined the time course of Na channel inactivation in clonal pituitary (GH3) cells by comparing records before and after the enzymatic removal of inactivation. The cells were subjected to whole-cell patch clamp, with papain included in the internal medium. Inactivation was slowly removed over the course of 10 min, making it possible to obtain control records before the enzyme acted. Papain caused a large (4-100x) increase in current magnitude for small depolarizations (near -40 mV), and a much smaller increase for large ones (approximately 1.5x at +40 mV). For technical reasons it was sometimes convenient to study outward INa recorded with no Na+ outside. The instantaneous I-V (IIV) curve in this condition was nonlinear before papain, and more nearly linear afterwards. The gNa-V curve after papain, obtained by dividing the INa-V curve by the IIV curve, was left-shifted by at least 20 mV and steepened. A spontaneous 5 10 mV left shift occurred in the absence of papain. The rate of the inactivation step was found to vary only slightly from -100 mV to +60 mV, based on the following evidence. (a) Before papain, inactivation rate saturated with voltage and was constant from +20 to +60 mV. (b) We activated the channels with a brief pulse, and studied the time course of the current on changing the voltage to a second, usually more negative level (Na+ present internally and externally). The time course of inactivation at each voltage was obtained by comparing control traces with those after inactivation was removed. When the 5-10-mV spontaneous shift was taken into account, inactivation rate changed by less than 10% from 100 to +60 mV. The data are considered in terms of existing models of the Na channel. PMID- 2552000 TI - Static and dynamic properties of synaptic transmission at the cyto-neural junction of frog labyrinth posterior canal. AB - The properties of synaptic transmission have been studied at the cyto-neural junction of the frog labyrinth posterior canal by examining excitatory postsynaptic potential (EPSP) activity recorded intraaxonally from the afferent nerve after abolishing spike firing by tetrodotoxin. The waveform, amplitude, and rate of occurrence of the EPSPs have been evaluated by means of a procedure of fluctuation analysis devised to continuously monitor these parameters, at rest as well as during stimulation of the semicircular canal by sinusoidal rotation at 0.1 Hz, with peak accelerations ranging from 8 to 87 deg.s-2. Responses to excitatory and inhibitory accelerations were quantified in terms of maximum and minimum EPSP rates, respectively, as well as total numbers of EPSPs occurring during the excitatory and inhibitory half cycles. Excitatory responses were systematically larger than inhibitory ones (asymmetry). Excitatory responses were linearly related either to peak acceleration or to its logarithm, and the same occurred for inhibitory responses. In all units examined, the asymmetry of the response yielded nonlinear two-sided input-output intensity functions. Silencing of EPSPs during inhibition (rectification) was never observed. Comparison of activity during the first cycle of rotation with the average response over several cycles indicated that variable degrees of adaptation (up to 48%) characterize the excitatory response, whereas no consistent adaptation was observed in the inhibitory response. All fibers appeared to give responses nearly in phase with angular velocity, at 0.1 Hz, although the peak rates generally anticipated by a few degrees the peak angular velocity. From the data presented it appears that asymmetry, adaptation, and at least part of the phase lead in afferent nerve response are of presynaptic origin, whereas rectification and possible further phase lead arise at the encoder. To confirm these conclusions a simultaneous though limited study of spike firing and EPSP activity has been attempted in a few fibers. PMID- 2551999 TI - Kinetic diversity of Na+ channel bursts in frog skeletal muscle. AB - Individual Na+ channels of dissociated frog skeletal muscle cells at 10 degrees C fail to inactivate in 0.02% of depolarizing pulses, thus producing bursts of openings lasting hundreds of milliseconds. We present here a kinetic analysis of 87 such bursts that were recorded in multi-channel patches at four pulse potentials. We used standard dwell-time histograms as well as fluctuation analysis to analyze the gating kinetics of the bursting channels. Since each burst contained only 75-150 openings, detailed characterization of the kinetics from single bursts was not possible. Nevertheless, at this low kinetic resolution, the open and closed times could be well fitted by single exponentials (or Lorentzians for the power spectra). The best estimates of both the open and closed time constants produced by either technique were much more broadly dispersed then expected from experimental or analytical variability, with values varying by as much as an order of magnitude. Furthermore, the values of the open and closed time constants were not significantly correlated with one another from burst to burst. The bursts thus expressed diverse kinetic behaviors, all of which appear to be manifestations of a single type of Na+ channel. Although the opening and closing rates were dispersed, their average values were close to those of alpha m and 2 beta m derived from fits to the early transient Na+ currents over the same voltage range. We propose a model in which the channel has both primary states (e.g., open, closed, and inactivated), as well as "modes" that are associated with independent alterations in the rate constants for transition between each of these primary states. PMID- 2552001 TI - Glutaraldehyde fixation of the cAMP-dependent Na+/H+ exchanger in trout red cells. AB - It has been shown that the addition of a beta-adrenergic catecholamine to a trout red blood cell suspension induces a 60-100-fold increase of sodium permeability resulting from the activation of a cAMP-dependent Na+/H+ antiport. Subsequent addition of propranolol almost instantaneously reduces the intracellular cAMP concentration, and thus the Na permeability, to their basal values (Mahe et al., 1985). If glutaraldehyde (0.06-0.1%) is added when the Na+/H+ exchanger is activated after hormonal stimulation, addition of propranolol no longer inhibits Na permeability: once activated and fixed by glutaraldehyde, the cAMP dependence disappears. Glutaraldehyde alone causes a rapid decrease in the cellular cAMP concentration. In its fixed state the antiporter is fully amiloride sensitive. The switching on of the Na+/H+ exchange by cAMP is rapidly (2 min) followed by acute but progressive desensitization of the exchanger (Garcia-Romeu et al., 1988). The desensitization depends on the concentration of external sodium, being maximal at a normal Na concentration (145 mM) and nonexistent at a low Na concentration (20 mM). If glutaraldehyde is added after activation in nondesensitizing conditions (20 mM Na), transfer to a Na-rich medium induces only a very slight desensitization: thus the fixative can "freeze" the exchanger in the nondesensitizing conformation. NO3- inhibits the activity of the cAMP dependent Na+/H+ antiporter of the trout red blood cell (Borgese et al., 1986). If glutaraldehyde is added when the cells are activated by cAMP in a chloride containing medium, the activity of the exchanger is no longer inhibited when Cl- is replaced by NO3-. Conversely, after fixation in NO3- medium replacement of NO3 by Cl- has very little stimulatory effect. This indicates that the anion dependence is not a specific requirement for the exchange process but that the anion environment is critical for the switching on of the Na+/H+ exchanger and for the maintenance of its activated configuration. PMID- 2552003 TI - Identification of virus-specific polypeptides by monoclonal antibodies against serotype 2 Marek's disease virus. AB - A total of 41 antibody-secreting hybridoma cells against the HPRS24 strain of Marek's disease virus (MDV) type 2 (MDV2) have been isolated. Of these monoclonal antibodies (MAbs), 24 were found by immunofluorescence tests to react specifically with MDV2-infected cells, but not MDV type 1 (MDV1)- or herpesvirus of turkeys (HVT)-infected cells, while eight reacted with MDV1- or MDV2-infected cells and nine with MDV1-, MDV2- or HVT-infected cells. By using these MAbs, seven classes of MDV type-specific or cross-reactive polypeptides were characterized by immunoprecipitation followed by SDS-PAGE. Among them, a 28K/32K glycoprotein differed from the previously identified gA and gB. The 28K/32K glycoprotein was found on the surface of MDV2-infected cells and in the cytoplasm by an immunofluorescence test with MAbs. In addition, a cross-reactive polypeptide of 25K/29K was also detected in MDV1-infected cells with MAbs reactive with the 28K/32K glycoprotein of MDV2. PMID- 2552002 TI - Localization of human papillomavirus type 16 DNA using the polymerase chain reaction in the cervix uteri of women with cervical intraepithelial neoplasia. AB - The localization of human papillomavirus type 16 (HPV-16) DNA throughout the cervix uteri of women with cervical intraepithelial neoplasia (CIN) was studied by utilizing the polymerase chain reaction technique directly on histologically defined sections of paraffin-embedded cervical tissue obtained by conizations. HPV-16 DNA was detected only in the sections that contained CIN lesions and/or koilocytes. No HPV-16 DNA was detected in sections that contained only normal epithelium. This is in accordance with HPV-16 playing a role in the development of CIN lesions. PMID- 2552004 TI - Immunoblot analysis of the antibody response to murine cytomegalovirus in genetically resistant and susceptible mice. AB - The murine of human cytomegalovirus infection was employed to analyse the kinetics of antibody production to murine cytomegalovirus (MCMV) structural and immediate early (IE) polypeptides following MCMV infection of genetically resistant and susceptible strains of mice. A total of 22 structural and six non structural. IE proteins were identified. Analysis of immunoblots by densitometry identified four patterns of antibody reactivity to MCMV structural polypeptides during primary and secondary antibody responses over a period of 5 weeks post infection (p.i.). Firstly, antibodies were strongly reactive with an 83K protein soon after infection, with levels which decreased with time: antibodies to a second group of viral proteins were also recognized soon after infection, but consistent levels of reactivity were maintained. Viral proteins that were recognized beyond day 14 p.i. or following a second MCMV infection formed the third group; the fourth consisted of viral proteins that were detected at variable times p.i. by antisera different mouse strains. The kinetics and intensity of the antibody response to individual viral proteins were influenced by virus dose, time p.i. and by a second MCMV infection. In addition, the genetic constitution of the host influenced the antibody response to MCMV proteins both quantitatively and qualitatively. In particular, sera from mice possessing C57BL but not BALB/c genes detected a 56K Mr viral during seroconversion. Antibody reactivity to this protein was shown to segregate among sera from CXB mice, with a strain distribution pattern which indicated a linkage with the b locus on chromosome 4. Finally, expression of the 56K protein was detected in B10. BR but not BALB/c embryo fibroblasts in vitro, with expression being a dominant trait in (BALB/c x B10.BR) F1 embryo fibroblasts. Thus, host genes may influence the expression of this structural viral protein. PMID- 2552006 TI - Vesicular stomatitis virus RNA replication: a role for the NS protein. AB - Synthesis of the vesicular stomatitis virus nucleocapsid (N) protein is required for viral RNA replication. The observation that the N protein forms a rapidly sedimenting species in the absence of other viral proteins and the description of complexes of N protein with NS protein led to the proposal that NS protein binds to N protein to prevent it from self-associating. We tested this model by analysing the physical properties of N protein synthesized alone in an in vitro replication system as compared to N protein synthesized in the presence of the NS protein. These findings were correlated with the ability of the N protein, synthesized under both conditions, to support replication. N protein synthesized at low concentrations in the absence of other viral proteins sedimented at 4S on glycerol gradients and was capable of supporting RNA replication. In contrast, synthesis of increasing concentrations of N protein resulted in formation of a rapidly sedimenting species of N protein which had the physical properties of a protein-protein aggregate and which failed to support RNA replication. Co synthesis of the NS protein with N protein both prevented the concentration dependent aggregation of N and restored the ability of high concentrations of N protein to support RNA replication. PMID- 2552005 TI - Analysis of reassortment and superinfection during mixed infection of Vero cells with bluetongue virus serotypes 10 and 17. AB - The reassortment of genome segments during mixed infection of Vero cells with bluetongue virus (BTV) serotypes 10 and 17 was investigated, using non-selective conditions for analysis of the progeny of mixed infections. Reassortment was found to be an early event in the BTV replication cycle, indicating that progeny BTV genomes undergo a single round of reassortment. Non-random segregation of individual genome segments was observed in crosses at equal multiplicity of infection, and was confirmed in crosses performed at unequal multiplicity. Asynchronous infections showed that superinfection exclusion resulted in the failure of the superinfecting virus to contribute genome segments to reassortants if the second virus followed the first by more than 4 h. The significance of these results for the evolution and epidemiology of BTV is discussed. PMID- 2552007 TI - Altered ATP function of a vesicular stomatitis virus mutant detected by kinetic analysis of the transcriptase using phosphorylated ribavirin. AB - We have studied the effect of phosphorylated ribavirin on the vesicular stomatitis virus (VSV) in vitro polymerase reaction by analysis of kinetic data obtained by varying the concentration of nucleoside triphosphates. The wild-type VSV had previously shown a competitive inhibition with the four natural nucleoside triphosphates with the use of ribavirin diphosphate (RDP) or ribavirin triphosphate (RTP). In contrast, when RDP (or RTP) was added to a transcription assay system using the polR1 mutant of VSV, a non-competitive or mixed type of inhibition was observed when the concentration of ATP was varied. Our results indicate that polR1 has an altered ATP function in addition to the previously described phenotypic characteristics of this mutant, which include synthesis of readthrough products of the leader/nucleocapsid (N) gene junction and a decreased ATP requirement for transcription. We have also studied CsCl-purified in vitro transcription products by primer-extending leader or N mRNA transcripts and found that the ratio of leader/N mRNA for VSV polR1 (1.3:1) was lower than values obtained previously for wild-type (3.7:1). PMID- 2552009 TI - Nucleotide sequence of high-passage hepatitis A virus strain HM175: comparison with wild-type and cell culture-adapted strains. AB - The nucleotide sequence of cDNA from a high-passage, cell culture-adapted variant of hepatitis A virus strain HM175 was compared with the previously determined sequences of wild-type virus and two other cell culture-adapted variants. A total of 42 nucleotide changes were detected when the sequence was compared with wild type virus. Five of these changes were common to all cell culture-adapted strains and a further two changes were shared by the strains that had experienced the greatest number of cell culture passages. The mutations were distributed throughout the genome coding for amino acid substitutions in regions 2B, 2C and 3D with silent changes in 1C and the 5' non-coding region. The possible relevance of these mutations to cell culture adaptation and attenuation is discussed. PMID- 2552008 TI - Sequences in the 5' non-coding region of human rhinovirus 14 RNA that affect in vitro translation. AB - A subgenomic cDNA clone from human rhinovirus 14 (HRV-14), comprising the 5' non coding region and the first 1182 nucleotides of the coding sequence, has been inserted into a vector under the control of the T7 promoter, and RNA was transcribed. Deletions in the 5' non-coding sequence modulated viral polyprotein synthesis significantly in a reticulocyte lysate system. Removal of the first 491 nucleotides had little effect, but deletion of a further 55 nucleotides (491 to 546) significantly increased the efficiency of the translation process. Further deletion to nucleotide 621 almost abolished translation, suggesting an essential role for the 546 to 621 nucleotide sequence. The efficiency of the translation process can also be influenced by the addition of ribosomal salt wash prepared from uninfected HeLa cells. PMID- 2552010 TI - Kunjin virus isolates of Australia are genetically homogeneous. AB - The genomes of 22 isolates of Kunjin virus (KUN) from Australia were characterized and compared using RNase T1 oligonucleotide fingerprinting. The results show that all isolates belonged to one topotype, the distribution of which covered the entire Australian continent. This finding is similar to that of Murray Valley encephalitis virus, but in contrast to the results reported for some other flaviviruses such as Saint Louis encephalitis virus. PMID- 2552011 TI - Metal complexes of antiinflammatory drugs. Part VII: Salsalate complex of copper(II). AB - The preparation and properties of the Cu(II) complex Cu(SAS)2.H2O are reported for the antiinflammatory drug Salsalate (SAS). The diffuse reflectance spectra and magnetic moments are consistent with a dinuclear structure as found for [Cu(aspirinate)2(H2O)]2. The Cu(II) complex exhibits an increased superoxide dismutase activity compared with the parent drug molecule in the nitroblue tetrazolium assay. PMID- 2552012 TI - Free radical intermediates formed during the oxidation of cyanide by horseradish peroxidase/H2O2 as detected with nitroso spin traps. AB - Aqueous solutions of cyanide react with hydrogen peroxide/horseradish peroxidase and form the cyanyl radical, which can be trapped by 2-methyl-2-nitrosopropane (t nitrosobutane, tNB) at pH 9.8. At lower pH a variety of radical adducts are formed; at higher pH, the main product was the spin adduct of the formamide radical with tNB. The use of deuterated tNB and 15N-labeled potassium cyanide allowed the observation of the very small nitrogen coupling of this radical adduct. Experiments using 3,5-dibromo-4-nitrosobenzenesulfonic acid (DBNBS) as the spin trap yielded only the formamide radical adduct, which was identified by an independent synthesis starting from formamide. Both hydrogen splittings of its amino group could be resolved using deuterated DBNBS as the spin trap. PMID- 2552013 TI - Kinetics of heme octapeptide (microperoxidase-8; MP-8) formation studied by high pressure liquid chromatography (HPLC) monitoring of the peptic and tryptic hydrolysis of horse heart cytochrome-c. AB - The kinetics of the sequential peptic and tryptic hydrolysis of cytochrome-c to give the heme-peptides microperoxidase-11 (MP-11) and -8 (MP-8), respectively, has been investigated by high performance liquid chromatography (HPLC), and we demonstrate that MP-8 can be prepared from cytochrome-c to the point of lyophilization within 4 hr. PMID- 2552014 TI - Cholinergic synaptic vesicles contain a V-type and a P-type ATPase. AB - Fifty to eighty-five percent of the ATPase activity in different preparations of cholinergic synaptic vesicles isolated from Torpedo electric organ was half inhibited by 7 microM vanadate. This activity is due to a recently purified phosphointermediate, or P-type, ATPase, Acetylcholine (ACh) active transport by the vesicles was stimulated about 35% by vanadate, demonstrating that the P-type enzyme is not the proton pump responsible for ACh active transport. Nearly all of the vesicle ATPase activity was inhibited by N-ethylmaleimide. The P-type ATPase could be protected from N-ethylmaleimide inactivation by vanadate, and subsequently reactivated by complexation of vanadate with deferoxamine. The inactivation-protection pattern suggests the presence of a vanadate-insensitive, N-ethylmaleimide-sensitive ATPase consistent with a vacuolar, or V-type, activity expected to drive ACh active transport. ACh active transport was half-inhibited by 5 microM N-ethylmaleimide, even in the presence of vanadate. The presence of a V-type ATPase was confirmed by Western blots using antisera raised against three separate subunits of chromaffin granule vacuolar ATPase I. Both ATPase activities, the P-type polypeptides, and the 38-kilodalton polypeptide of the V type ATPase precisely copurify with the synaptic vesicles. Solubilization of synaptic vesicles in octaethyleneglycol dodecyl ether detergent results in several-fold stimulation of the P-type activity and inactivation of the V-type activity, thus explaining why the V-type activity was not detected previously during purification of the P-type ATPase. It is concluded that cholinergic vesicles contain a P-type ATPase of unknown function and a V-type ATPase which is the proton pump. PMID- 2552015 TI - Postmortem degradation alters fluorographic labeling patterns and affinities of benzodiazepine binding proteins. AB - To investigate the effect of endogenous proteolysis on the molecular weights of the benzodiazepine binding proteins, brains of trout, chicken, and rat were removed immediately after death and stored at room temperature for various periods of time before they were frozen. Photoaffinity labeling of membranes with [3H]flunitrazepam, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography, revealed proteolytic fragments of 47K in trout, chicken, and rat. The proteolysis set in rapidly after death. Seemingly in parallel with the degradation observed fluorographically, the affinity for [3H]flunitrazepam increased without systematic changes in receptor density. The degradation pattern was not identical to that of the photolabeled trypsinized benzodiazepine binding proteins. The endogenous proteolytic fragments were deglycosylated in two steps. In conclusion, proteolytic effects must be taken into account when interpreting labeling patterns and binding parameters. PMID- 2552016 TI - Solubilization and characterization of sigma-receptors from guinea pig brain membranes. AB - The sigma-receptor, a distinct binding site in brain tissue that may mediate some of the psychotomimetic properties of benzomorphan opiates and phencyclidine, has been solubilized using the ionic detergent sodium cholate. Binding assays were performed with the solubilized receptor using vacuum filtration over polyethyleneimine-treated glass fiber filters. The pharmacological specificity of the solubilized binding site for sigma-receptor ligands is nearly identical to the membrane-bound form of the receptor, with the order of potencies for displacement of the selective sigma-ligand [3H]di-o-tolylguanidine ([3H]DTG) closely correlated. The stereoselectivity for (+)-benzomorphan opiate enantiomers was retained by the solubilized receptor. The soluble receptor retained high affinity for binding of [3H]DTG (KD = 28 +/- 0.5 nM) and (+)-[3H]3-(3 hydroxyphenyl)-N-(1-propyl)piperidine [(+)-[3H]3-PPP] (KD = 36 +/- 2 nM). Photoaffinity labeling of the solubilized receptor by [3H]p-azido-DTG, a sigma selective photoaffinity label, resulted in labeling of a 29-kilodalton polypeptide identical in size to that labeled in intact membranes. Estimation of the Stokes radius of the [3H]DTG binding site was obtained by Sepharose CL-6B chromatography in the presence of 20 mM cholate and calculated to be 8.7 nm. This value was identical to the molecular size found for the binding sites of the sigma-selective ligands (+)-[3H]3-PPP and (+)-[3H]SKF-10,047, supporting the hypothesis that all three ligands bind to the same macromolecular complex. PMID- 2552017 TI - Presynaptic kappa-opioid receptors on noradrenergic nerve terminals couple to G proteins and interact with the alpha 2-adrenoceptors. AB - Stimulation-induced noradrenaline (NA) release in rabbit hippocampus is inhibited by activation of presynaptic alpha 2-adrenoceptors and kappa-opioid receptors. The purpose of the present study was to investigate (a) an interference between the alpha 2- and kappa-mechanisms, and (b) a coupling of the opioid receptors to pertussis toxin (PT)-sensitive guanine nucleotide-binding proteins (G proteins), as has been previously shown for the alpha 2-receptors. [3H]NA release from hippocampal slices was evoked by electrical field stimulation (360 pulses/3 Hz). Inhibition of stimulation-evoked NA release by the preferential kappa-receptor agonist ethylketocyclazocine (EKC) was increased in the presence of the alpha 2 adrenoceptor antagonist yohimbine (0.1 or 1.0 microM). When autoinhibition was completely removed, EKC (1 microM) almost abolished transmitter release. Pretreatment of hippocampal tissue with either PT (8 micrograms/ml; 18 h) or N ethylmaleimide (NEM) (30 microM; 30 min), which has been shown to alkylate PT substrates, diminished the EKC-produced inhibition of NA release. The kappa mechanism was still impaired by these compounds when the alpha 2-receptors were blocked with yohimbine. An effect of NEM on the active site of the kappa-receptor seems to be unlikely, because NEM diminished the EKC-induced inhibition of release irrespective of whether or not the opioid receptor was occupied by EKC during exposure to NEM. The present results suggest an interference of both alpha 2- and kappa-opioid receptor-coupled signal transduction possibly through competition for a common pool of G proteins. PMID- 2552018 TI - Regulation of the gamma-aminobutyric acidA receptor by gamma-aminobutyric acid levels within the postsynaptic cell. AB - Synaptosomes and synaptoneurosomes were prepared from rat cerebral cortex. Comparison of the amino acid levels in the two types of organelles and of the effects of gabaculine thereon indicated that the neurosome portion of synaptoneurosomes constituted the major influencing component of the organelles. Administration to rats of inhibitors of gamma-aminobutyric acid (GABA) degradation, such as gabaculine and L-cycloserine, resulted in elevated GABA levels in synaptoneurosomes and a decrease in muscimol-stimulated Cl- up-take by the organelles. Addition of gabaculine directly to the incubation medium for the uptake assay had no effect on the Cl- transport. In contrast, administration to rats of isonicotinic acid hydrazide, an inhibitor of GABA synthesis, decreased the GABA level in synaptoneurosomes and increased the muscimol-stimulated Cl- uptake by the organelles. Although the evidence is not unequivocal, it does support the concept of GABA released from nerve endings being taken up by the postsynaptic cell, from where it exerts a regulatory influence on the functioning of the GABA receptor/ion channel complex. PMID- 2552019 TI - The affinity of bone to hydroxyapatite and alumina in experimentally induced osteoporosis. AB - The authors studied how bone reacts to hydroxyapatite (HAP) and alumina (Al2O3) in osteoporosis experimentally induced in a rat animal model by resection of the sciatic nerve. Sixteen and 32 weeks after resection of the sciatic nerve, implants of HAP and Al2O3 were inserted into rat tibias. The reaction of bone to HAP was compared with its reaction to Al2O3, which is bioinert, as a control. The results were analyzed using the affinity index (the length of bone directly opposed to the implant without an intervening fibrous membrane/total length of the bone-implant interface x 100%). In the rats left for 16 weeks after resection of the sciatic nerve the index values 24 weeks after insertion of HAP and Al2O3 were 96.9 +/- 2.4% (N = 8) and 75.2 +/- 22.2% (N = 8), respectively. The index value of HAP was 97.2 +/- 2.6% (N = 12) and of Al2O3 was 86.9 +/- 15.4% (N = 12) at 24 weeks after insertion, when the rats were first left for 32 weeks, after sciatic nerve resection. Bone was demonstrated to have a superior affinity to HAP, compared with Al2O3 (P less than .05). PMID- 2552020 TI - Soluble class I antigens in serum and CSF of patients with varicella-zoster virus meningitis. AB - Soluble class I antigens (sHLA) are secreted by lymphocytes upon activation in vitro. The intrathecal synthesis (ITS) of these molecules has been studied in patients with the varicella-zoster virus (VZV) meningitis. In this paper we describe a sHLA index IH = (CSF sHLA/serum sHLA): (CSF albumin/serum albumin) which is expected to increase only when sHLA is synthesised within the central nervous system (CNS). The IH is elevated in the first week of meningitis, when antibody synthesis is still low, and decreases thereafter. We think IH is an index of early lymphocyte activation within the CNS. The relation of these findings with previous in vitro studies is also discussed. PMID- 2552021 TI - Cerebral venous thrombosis and dual primary infection with human immunodeficiency virus and cytomegalovirus. PMID- 2552022 TI - Delayed somatosensory evoked potentials in pernicious anaemia with intact peripheral nerves. PMID- 2552023 TI - Corticospinal potentials after transcranial stimulation in humans. AB - The descending volley evoked in humans by transcranial electrical stimulation of the scalp was recorded with epidural and spinal electrodes. It consisted of an early wave, which increased in amplitude and decreased in latency when the strength of the stimulus was increased. The mean conduction velocity of the early wave was 66, SD 2.5 m/s. At high stimulus intensity this wave was followed by later and smaller waves, which travel at the same speed as the initial potential. The recovery cycle of the descending volley was studied by delivering paired cortical stimuli at time intervals ranging from 0.5 to 10 ms. The early wave evoked by the test stimulus recovered to about 50% at a 1 ms interval and to 100% at a 3.5 ms interval. The later waves could not be tested at short time intervals but with time intervals longer than 3.5 ms they recovered to 100%. It is suggested that the initial and later waves after scalp stimulation are equivalent to the D and I waves seen in animal experiments. PMID- 2552025 TI - Monoclonal antibody to a human salivary gland adenocarcinoma cell line: augmentation of antibody-dependent cell-mediated cytotoxicity activity by streptococcal preparation OK-432 in human salivary gland adenocarcinoma-bearing nude mice given the antibody. AB - An IgG2a mouse monoclonal antibody (MoAb) to the human salivary gland adenocarcinoma cell line HSG, 5B/10, has been generated in our laboratory. In the current study, the antitumor effects mediated by MoAb 5B/10 in human salivary gland adenocarcinoma (HSG)-bearing nude mice or the antibody-dependent cell mediated cytotoxicity (ADCC) against HSG cells using human peripheral blood mononuclear cells (PBMC) as effector cells were examined. In addition, effects of the streptococcal preparation OK-432 on the growth of HSG tumors or on the MoAb 5B/10-mediated cellular cytotoxicity were studied. MoAb 5B/10 mediated an ADCC reaction against HSG cells that were insensitive to NK cells but not to the reaction of antibody and complement-mediated cytotoxicity. The coexistence of MoAb 5B/10 and OK-432 caused marked augmentation of cytotoxic effects. Treatment of OK-432-stimulated PBMC with antiasialo Gm1 antiserum plus complement but not with silica particles resulted in a significant decrease of cytotoxic effects as compared with relevant controls. the nude mice inoculated intraperitoneally with HSG cells and treated with MoAb 5B/10, OK-432, or (especially) a combination of the two had significantly prolonged survival times as compared with untreated controls. Moreover, spleen cells from the tumor-bearing mice treated with OK-432 alone or a combination of MoAb 5B/10 and OK-432 were found to carry high levels of effector cell activity in the MoAb 5B/10-mediated cytotoxicity assay using HSG cells as targets. PMID- 2552024 TI - Combined HIV-CMV encephalitis presenting with brainstem signs. AB - Two cases of combined HIV-CMV encephalitis are described. One presented with a sixth nerve palsy and a tetraparesis, the other with an internuclear ophthalmoplegia. Pathologically brain stem involvement was predominantly due to CMV. PMID- 2552026 TI - Cytokine augmentation of human immunodeficiency virus type 1 (HIV-1) gp120 specific cellular cytotoxicity. AB - Currently available anti-human immunodeficiency virus type 1 (HIV-1) agents such as azidothymidine can prevent de novo virus infection in vitro but lack significant activity against chronically infected cells. Our laboratory has recently described glycoprotein (gp)120-specific cell mediated cytotoxicity (CMC) present in HIV-1-seropositive individuals that is capable of destroying virally infected cells. As a means of potentially eliminating persistent reservoirs of HIV-1, we examined the ability of various cytokines to augment preexisting gp120 specific CMC activity of peripheral blood mononuclear cells obtained from early disease patients. We found that interferon-gamma alone had no effect on gp120 cellular reactivity; however, the combination of interferon-gamma plus IL-2 produced enhancement beyond that of IL-2 alone. PMID- 2552027 TI - Herpes simplex virus (HSV) DNA in microglial nodular brainstem encephalitis. AB - Thirty-four brains with microglial nodular brain stem encephalitis were retrospectively investigated for herpes simplex virus (HSV) and cytomegalovirus (CMV) by in situ hybridization (ISH) with biotinylated cDNA probes, and by immunocytochemistry with polyclonal and monoclonal antibodies on formalin fixed paraffin embedded serial tissue sections. In 16 cases (47%), HSV DNA was found by ISH in the nuclei of neurons in microglial nodules or in the adjacent parenchyma of the brainstem, and more rarely at various cerebellar and telencephalic sites. None of the 34 cases was labeled for CMV DNA and none revealed HSV or CMV antigens. Ten control brains without microglial nodules were not labeled. This study suggests an HSV etiology for many cases with microglial nodular brainstem encephalitis. PMID- 2552028 TI - Rationale for surgery as the first step in the multimodality treatment of small cell lung cancer (SCLC). AB - The role of surgery has to be reconsidered in combination with chemo/radiotherapy for patients with SCLC at early stages, determined by the pTNM-staging system. This combined multimodality treatment can be seen as a model-like example for the most effective progress gained during the last 20 years. Surgery followed by chemotherapy is used in ongoing cooperative studies. In other trials, chemotherapy is followed by surgery for responding patients. It seems reasonable to expect that after appropriate observation times the comparison of these ongoing trials would most probably lead to the conclusion that for stage I, II surgery should be the first step followed by chemo- and radiotherapy, whereas for stage III debulking chemotherapy should be given to distinguish responding patients, who may thereafter receive adjuvant surgery. PMID- 2552030 TI - In vitro and in vivo efficacy of YTR-830H and piperacillin combinations versus beta-lactamase-producing bacteria. AB - YTR-830H, a beta-lactamase inhibitor, is a non-amino penicillanic sulfone. In vitro synergistic activity with piperacillin was determined for 226 beta lactamase producing clinical cultures. Combination of piperacillin: YTR in ratios of 2:1, 4:1, and 8:1 were highly effective vs Escherichia coli, Proteus, Providencia, Morganella, Staphylococcus, and Bacteroides. Minimum inhibitory concentrations (MICs) of piperacillin were reduced from the resistant to susceptible range. The higher ratios were less effective vs Enterobacter, Serratia, and Citrobacter. YTR-830H was not antagonistic with piperacillin. Combinations of 2:1, 4:1, and 8:1 increased the therapeutic effectiveness of piperacillin 8 - to 36 - fold against acute lethal infections produced in mice with piperacillin-resistant Escherichia coli, Klebsiella pneumoniae, Morganella morganii, and Staphylococcus aureus. PMID- 2552029 TI - In vitro activity of YTR 830. AB - YTR 830, now known as tazobactam, is a new penicillanic acid sulfone beta lactamase inhibitor. The in vitro activity of YTR 830 combined with various penicillins was determined and compared to that of clavulanate and sulbactam combined with the same agents. Combined with ampicillin or amoxicillin, all three inhibitors were active against beta-lactamase producing strains of Staphylococcus aureus, Haemophilus influenzae, Klebsiella, Citrobacter diversus, and all anaerobes except for Bacteroides fragilis homology group II. YTR 830 was also effective against Escherichia coli and indole-positive Proteus. The inhibitors had no effect against Enterobacter or Serratia. Overall, the activity of YTR 830 was comparable to that of clavulanate, and superior to that of sulbactam. PMID- 2552031 TI - The effectiveness of combination chemotherapy with cisplatinum and etoposide in the treatment of advanced non-small cell lung cancer. AB - Twenty-one patients with histologically proven advanced or disseminated non-small cell lung cancer were treated with cisplatinum 80 mg/m2 i.v. day 1 and etoposide (VP16) 80 mg/m2 i.v. day 1- greater than 3.15 patients were evaluable for response. One patient (6.6%) achieved a complete response, 4 (26.7%) a partial response and 6 (40.0%) a stabilization of disease. Four patients (26.7%) progressed. An improvement in performance status was obtained in more than 50% of cases. Responders had a mean survival of 345 + days, while non-responders 191.7 days. The treatment was generally well tolerated. In our opinion this combination regimen offers good palliation for patients affected by advanced and/or metastatic non-small cell lung cancer. PMID- 2552033 TI - Ca2+ and Na+ currents in developing skeletal myoblasts are expressed in a sequential program: reversible suppression by transforming growth factor beta-1, an inhibitor of the myogenic pathway. AB - We have analyzed the biophysical and developmental properties of Ca2+ and Na+ currents in C2 muscle cells, whose morphological and biochemical phenotype closely resembles differentiated skeletal muscle. Both fused and unfused C2 myocytes possessed: (1) membrane capacitance consistent with the presence of complex sarcotubular invaginations, (2) tetrodotoxin-sensitive Na+ channels, and (3) "fast" and "slow" Ca2+ channels that inactivated at holding potentials of -40 and -20 mV, respectively. Thus, the passive electrical properties, Na+ currents, and Ca2+ currents expressed in C2 cells each differed from those found in the nonfusing muscle cell line, BC3H1, and corresponded more precisely to characteristic findings observed in skeletal muscle fibers. In further contrast to BC3H1 cells, C2 muscle also expressed "transient" Ca2+ channels similar to those reported in embryonic or neonatal skeletal muscle, which were detected within 12-24 hr of mitogen withdrawal, up to 60 hr before appearance of "fast" and "slow" currents. Na+ channels also were induced 12-24 hr after mitogen withdrawal. Unlike the "fast" and "slow" Ca2+ currents, which were maximally expressed at 8-14 d of serum withdrawal, "transient" Ca2+ channels became down regulated upon prolonged differentiation (as found in postnatal skeletal muscle in vivo) and were no longer expressed at 14 d. Despite their divergent kinetic and developmental properties, all components of Ca2+ and Na+ current in C2 myocytes were suppressed reversibly in the presence of transforming growth factor beta-1, a purified growth factor that inhibits the myogenic phenotype. The results indicate that fusion is not essential for skeletal myoblasts to produce developmentally regulated voltage-gated channels that resemble those of intact muscle and demonstrate that the formation of diverse Ca2+ and Na+ channels can be mediated by a single peptide that affects the myogenic pathway. PMID- 2552032 TI - Sigma receptors regulate contractions of the guinea pig ileum longitudinal muscle/myenteric plexus preparation elicited by both electrical stimulation and exogenous serotonin. AB - Sigma receptors are specific, highly localized binding sites in limbic and sensorimotor structures of the brain that interact with many psychotropic drugs. These agents include the psychotomimetic benzomorphan opiates, the psychotomimetic drug phencyclidine and its analogs, as well as numerous typical and atypical antipsychotics such as haloperidol, chlorpromazine, and the novel drugs BMY 14802 and rimcazole. So far, no physiological function has been assigned to these binding sites. We have synthesized a number of novel sigma receptor-active drugs derived from the selective sigma ligand N,N'-di(o tolyl)guanidine (DTG). DTG and its congeners were found to inhibit contractions of the guinea pig ileal longitudinal muscle/myenteric plexus (LMMP) preparation evoked by electrical stimulation. In addition, the sigma ligands noncompetitively antagonized contractions of the LMMP preparation evoked by serotonin (5-HT). The 5-HT-evoked contractions were found to be largely due to 5-HT's activation of 5 HT3 receptors to release ACh. The activity of DTG congeners in inhibiting electrically or 5-HT-evoked contractions of the LMMP highly correlated with their potency to inhibit binding of both 3H-DTG and (+)3H-3-PPP [3(3-OH-phenyl)-N-(1 propyl)piperidine] to sigma receptors in guinea pig brain homogenates. Two DTG congeners that did not bind to sigma receptors also showed no activity in the bioassay. Many other (but not all) sigma receptor ligands showed a high correlation between their potency to inhibit electrically evoked contractions of the LMMP and their sigma receptor binding affinity. The benzomorphans (+)SKF 10,047 and (+)cyclazocine potentiated electrically evoked contractions of the LMMP. Sigma ligands also inhibited the contractions of the LMMP in the presence of the opiate antagonist naloxone and in preparations in which opioid receptors had been inactivated by treatment with the irreversible opiate antagonist beta chlornaltrexamine. Control experiments suggested that the sigma ligands act via a neuronal mechanism to inhibit ACh release evoked by electrical stimulation or by stimulation with 5-HT. These results suggest that there are functional sigma receptors on cholinergic nerve terminals or within the myenteric plexus and that these receptors can inhibit stimulated ACh release through an opioid receptor independent mechanism. However, sigma receptor activation in the ileum has the same effect on ACh release as activation of naloxone-sensitive opioid receptors. The LMMP may be an in vitro bioassay system for characterizing the mechanism of action of sigma receptors and for determining the biological efficacy of drugs known to bind to sigma receptors in radioligand binding assays. PMID- 2552034 TI - Expression of growth-associated protein B-50 (GAP43) in dorsal root ganglia and sciatic nerve during regenerative sprouting. AB - Recently it has been shown that B-50 is identical to the neuron-specific, growth associated protein GAP43. The present study reports on the fate of B-50/GAP43 mRNA and B-50/GAP43 protein, determined by radioimmunoassay, in a rat model of peripheral nerve regeneration (sciatic nerve crush) over a period of 37 and 312 d, respectively. Moreover, the effects of repeated subcutaneous injection of the neurotrophic peptide Org.2766 (an ACTH4-9 analog) and of a conditioning lesion on B-50/GAP43 protein levels in the regenerating nerve and dorsal root ganglia (DRG) were investigated. Both treatments enhanced the functional recovery as evidenced by a foot-flick withdrawal test. Immunocytochemical analysis using antineurofilament antibodies revealed a peptide-induced increase in the number of outgrowing sprouts in the sciatic nerve. Both the peptide and the conditioning lesion amplified the crush lesion-induced increase in B-50 protein content in the nerve as determined by radioimmunoassay. B-50 protein levels seem to correlate proportionally with the number of sprouts. In the DRG of the crushed sciatic nerve, the time course of B-50 expression was studied. B-50 mRNA was quantified from Northern blots. A linear increase up to 10 times the basal level of B-50 mRNA was observed 2 d postsurgery, followed by a gradual decline to normal levels at day 37. The first significant rise in B-50 mRNA level became apparent between 8 and 16 hr after placement of the crush lesion. The first significant rise in B 50 protein level occurred 40 hr after the crush lesion, reaching a plateau of 3 times the basal level between day 6 and 20. B-50 protein levels in DRG cell bodies remained elevated up to 60 d after crush, a period much longer than that observed for B-50 mRNA. Thus, during a later phase of peripheral axonal regeneration, the presence of B-50 appears to be prolonged, probably by an increase in half-life and not so much by enhanced transcription. Treatment with Org.2766 did not affect the B-50/GAP43 levels in DRG cell bodies during the first 6 d following crush. Conditioning lesion resulted in a DRG B-50/GAP43 protein amount at the same level as in rats 14 d after the test lesion. B-50/GAP43 levels in DRG are probably influenced by the rapid axonal transport of the protein, as has been reported by others. PMID- 2552035 TI - In situ protein phosphorylation in hippocampal tissue slices. AB - We have studied the subcellular distribution of phosphoproteins in intact hippocampal slices and examined factors that regulate their phosphorylation and dephosphorylation in situ. The presence of Ca2+ in slice equilibration and prelabeling buffers and high-K+-induced depolarization markedly increased 32Pi incorporation into endogenous proteins. Ca2+-stimulatory effects were significantly reduced by Ca2+-channel blockers and the calmodulin antagonist W 13. Certain proteins were dephosphorylated in situ, and their dephosphorylation was dependent on both Ca2+ and depolarization. A number of proteins phosphorylated in situ was similar to those previously characterized in synaptic fractions phosphorylated in vitro. Many phosphoproteins were identified on the basis of molecular weight, isoelectric point, immunoreactivity, and phosphopeptide mapping; these included the 87 kDa substrate of protein kinase C, synapsin I, the 50 and 60 kDa subunits of Ca2+/calmodulin-dependent protein kinase II (CKII), tubulin, B-50, the alpha-subunit of pyruvate dehydrogenase and myelin basic proteins. CKII phosphorylation in situ appeared similar but not identical to its in vitro counterpart. Phosphopeptide mapping analysis of in situ labeled substrate proteins indicated that cAMP-, Ca2+/calmodulin-, and Ca2+/phospholipid-dependent protein kinases were all active in slice preparations under basal conditions. Increased 32Pi labeling of hippocampal proteins following tissue depolarization appeared to be associated with increased activity of endogenous protein kinases since depolarization did not result in 32Pi-labeling of any new phosphoproteins. PMID- 2552037 TI - ARPP-21, a cyclic AMP-regulated phosphoprotein enriched in dopamine-innervated brain regions. II. Molecular cloning and nucleotide sequence. AB - A cDNA clone for the mRNA of bovine ARPP-21 (cAMP-regulated phosphoprotein, Mr = 21,000 as determined by SDS-PAGE) was isolated from a modified Okayama-Berg plasmid library. Transformed Escherichia coli colonies were screened by in situ colony hybridization with 2 different oligonucleotide probes derived from the amino acid sequence of the bovine protein. Sequence analysis of the longest cDNA clone, pTKAI [2407 nucleotides plus a poly(A) tail], revealed a 267-nucleotide long coding region in agreement with the bovine ARPP-21 amino acid sequence (Williams et al., 1989). Southern blot analysis of total bovine genomic DNA raised the possibility that there may be 2 genes coding for ARPP-21. Northern blot analysis of total cellular RNA from bovine caudate nucleus and other brain regions demonstrated the existence of 2 major mRNA species, 2.5 and 1.0 kb in length, probably derived from use of alternate polyadenylation sites. There was a differential expression of these 2 mRNAs within the brain. Both ARPP-21 mRNAs were most abundant in the caudate nucleus, where the concentration of the protein is highly enriched. PMID- 2552036 TI - ARPP-21, a cyclic AMP-regulated phosphoprotein enriched in dopamine-innervated brain regions. I. Amino acid sequence of ARPP-21B from bovine caudate nucleus. AB - ARPP-21 (cAMP-regulated phosphoprotein, Mr = 21,000 as determined by SDS-PAGE) is a major cytosolic substrate for cAMP-stimulated protein phosphorylation in dopamine-innervated regions of the rat CNS. It has recently been purified to homogeneity from bovine caudate nucleus and characterized (Hemmings and Greengard, 1989). ARPP-21 is isolated as 2 isoforms, ARPP-21A and ARPP-21B. The amino acid sequence of purified bovine ARPP-21B has now been determined by gas phase sequencing. The S-14C-carboxymethylated protein was subjected to enzymatic cleavage with trypsin, chymotrypsin, subtilisin, and endoproteinase Lys-C. The resulting peptides were purified by high-performance liquid chromatography, and selected peptides were subjected to amino acid analysis and/or amino acid sequencing by automated Edman degradation. ARPP-21B consists of a single NH2 terminal blocked polypeptide chain of 88 residues, with a calculated molecular mass of 9561 Da, including an NH2-terminal acetyl group inferred by deblocking with an acylaminopeptidase. This molecular mass is significantly lower than earlier estimates based on SDS-PAGE or hydrodynamic measurements. The seryl residue phosphorylated by cAMP-dependent protein kinase (Hemmings et al., 1989) is located at position 55. The molecule contains 1 cysteinyl residue, at position 71, and contains no methionyl, tyrosyl, phenylalanyl, tryptophanyl, or histidinyl residues. Determination of the primary structure of ARPP-21, one of several phosphoproteins localized to dopaminoceptive neurons in the basal ganglia, provides a framework for further investigations into the molecular mechanisms involved in dopaminergic neurotransmission. PMID- 2552038 TI - Localization and development of nerve growth factor-sensitive rat basal forebrain neurons and their afferent projections to hippocampus and neocortex. AB - In order to understand further the role of NGF in the development of NGF sensitive basal forebrain neurons and their afferent connections to the hippocampus and neocortex, we have used monoclonal antibody 192 IgG to detect and localize NGF receptors immunocytochemically in the developing rat brain. NGF receptor immunoreactivity (NGF-RI) is first visible at embryonic day 13 (E 13) in the ventrolateral telencephalic wall and follows a caudal-to-rostral gradient in its initial appearance. NGF-RI neuronal number and neuropil staining undergo substantial increases before birth, and extensive dendritic growth and increases in perikaryal size continue during the first 3 weeks of postnatal life. This growth and cellular differentiation, however, is followed in the fourth postnatal week and later by an apparent decrease in dendritic arborization and 50% shrinkage in the size of perikarya. Initial NGF-RI fiber outgrowth from immature basal forebrain neurons directed toward appropriate target fields is observed as early as E 15. The formation of a laminar pattern by septal axons in the hippocampal terminal fields and invasion of NB afferents into the cortex occur postnatally over a protracted time. In the hippocampus, NGF-RI is initially diffusely distributed, and wide bands of immature granule and pyramidal cells are almost devoid of immunoreactive fibers; however, with maturity, septal axon terminals become concentrated in narrow zones closely associated with the cellular layers. In the neocortex, early-arriving basal forebrain afferents accumulate in the intermediate zone underneath the darkly immunoreactive subplate before they enter The cortex. Dense subplate and transiently present, radially aligned fiber staining completely disappear in later postnatal week and are gradually replaced by specific axonal and terminal staining associated with NB afferents. The expression of NGF receptor in the subplate zone at the time afferents arrive and its subsequent disappearance with the specific terminal formation suggest that NGF receptor and concomitant accumulation of NGF in the subplate may act as a temporary target for the early-arriving basal forebrain afferents; ingrowing afferents may then be guided by radially oriented NGF-RI fibers to proper synaptic sites. PMID- 2552039 TI - Synaptic plasticity and learning: selective impairment of learning rats and blockade of long-term potentiation in vivo by the N-methyl-D-aspartate receptor antagonist AP5. AB - This paper reports a series of 5 experiments concerned with a possible role for N methyl-D-aspartate (NMDA) receptors in certain types of learning. The results show that chronic intraventricular infusion of the NMDA receptor antagonist D,L-2 amino-5-phosphonopentanoic acid (D,L-AP5) caused an impairment of spatial but not of visual discrimination learning in rats. Such selectivity of the learning impairment occurred despite widespread distribution of the drug throughout the CNS. AP5 sometimes caused a disturbance of sensorimotor function during learning, but one experiment addressing whether this disturbance could be responsible for the spatial learning impairment established that it was statistically independent. Another experiment showed that AP5 did not affect the retention of previously acquired spatial information. These behavioral effects were all obtained with a concentration of AP5 that, in a final study, was found to be sufficient to block hippocampal long-term potentiation (LTP) in vivo without affecting normal synaptic transmission. Taken together, these observations (1) implicate NMDA receptors in certain types of learning, and (2) extend recent work showing that saturation of LTP causes an anterograde spatial amnesia (McNaughton et al., 1986). A preliminary report of parts of this work has been published (Morris et al., 1986a). PMID- 2552040 TI - Compartments and the topography of leg afferent projections in Drosophila. AB - The legs of Drosophila are covered with mechanosensory bristles, innervated by sensory neurons that project to the CNS in a very orderly manner. We examined this afferent projection by staining the sensory neurons associated with identified bristles in wild-type, engrailed and scute flies. We observe that anterior neurons project to an anterior region of the ventral neuropil, while posterior neurons project to a more posterior region. We rule out that this difference depends on the compartment of origin of the receptors. Our results also argue against explanations based on other factors that might correlate to anterior/posterior position: peripheral organization of the leg nerve, competitive interactions, or differences in times of birth. We suggest that position itself is the primary determinant of this projection. PMID- 2552041 TI - Development of excitable membrane properties in mammalian sympathetic neurons. AB - Using the whole-cell patch-clamp recording technique, resting membrane potentials (RPs), action potential (AP) waveforms, and the properties of voltage-activated inward Na+ (lNa) and Ca2+ (lCa) currents and outward K+ (lA,lK) currents were examined in embryonic and neonatal rat superior cervical ganglion (SCG) neurons as a function of time during development in vivo and in vitro. The passive and active membrane properties of neonatal SCG cells examined less than or equal to 24 hr after isolation were similar to those described previously for adult SCG neurons and for neonatal SCG cells maintained for several weeks in culture. Since recordings were obtained within hours of cell dissociations, it is assumed that the results reflect the membrane properties of neonatal SCG neurons in vivo at the time of isolation. When neonatal cells were examined as a function of time (up to approximately 2-3 weeks) in vitro, neither RPs nor AP waveforms varied measurably. Although absolute (inward and outward) current amplitudes increased in cells maintained in vitro, in parallel with increases in cell size, no changes in the time- or voltage-dependent properties of the currents were observed. Similar results were obtained for cells isolated on or after embryonic day 18.5 (greater than or equal to E 18.5). The membrane properties of E 14.5-16.5 SCG cells examined less than or equal to 24 hr after isolation, in contrast, were significantly different: mean lCa density was higher, and APs were broader than in greater than or equal to E 18.5 cells, and, in addition, lA was absent in these cells. When E 14.5-16.5 cells were examined after approximately 1 week in vitro, lCa densities and AP waveforms were indistinguishable from those in greater than or equal to E 18.5 SCG neurons, and lA was present. These studies reveal that rat SCG neurons are electrophysiologically mature early in development. Even lA, which seems to be the last voltage-gated current to develop, appears well before birth. As the appearance of lA correlates with decreased membrane excitability and AP shortening, it seems likely that the development of lA either reflects or regulates a marked change in the overall input and output properties of developing sympathetic neurons. PMID- 2552042 TI - Identification of a cell-surface protein involved in PC12 cell-substratum adhesion and neurite outgrowth on laminin and collagen. AB - On substrata coated with laminin or native collagen (Types I/III), PC12 cells employ an active adhesion mechanism (i.e., one inhibited at low temperature, by azide or in the absence of divalent cations) to attach and extend neurites; on substrata coated with wheat germ agglutinin (WGA) or polylysine, by contrast, PC12 cells attach via a passive mechanism and fail to extend neurites (Turner et al., 1987). This paper reports the isolation of 2 monoclonal antibodies (3A3 and 1B1) that promote retraction of neurites extended on laminin and collagen. In studies of initial cell attachment, 3A3 inhibited active attachment to laminin or collagen but not passive attachment to WGA or polylysine, whereas 1B1 inhibited both active and passive attachment. The more potent of the antibodies, 3A3, precipitates 2 radioactive protein bands (of approximately 185 and 125 kDa) from 1% Nonidet P-40 extracts of metabolically labeled PC12 cells. The properties of these proteins suggest that the antigen recognized by 3A3 is a member of the integrin family of matrix receptors. The other monoclonal antibody, 1B1, reacts with many PC12 proteins, including both bands precipitated by 3A3. The available data strongly suggest that an integrin with specificity for both laminin and collagen mediates PC12 adhesion to the substratum at both the cell body and the neurite growth cone. PMID- 2552043 TI - Second messengers involved in the mechanism of action of bradykinin in sensory neurons in culture. AB - Application of bradykinin to neonatal rat dorsal root ganglion neurons caused a depolarization associated with an inward current and an increase in membrane conductance that was probably due to the opening of sodium channels. No hyperpolarization or outward current was detected. In addition, bradykinin increased the rate of 45Ca uptake into the neurons by a mechanism that was blocked by the dihydropyridine calcium channel antagonist nifedipine. Direct activation of protein kinase C (PKC) with phorbol esters mimicked the ability of bradykinin to depolarize the neurons and to increase the rate of 45Ca uptake. Down-regulation of PKC by prolonged treatment with phorbol esters and treatment of the cells with staurosporine, which inhibits PKC, blocked both bradykinin- and phorbol ester-induced 45Ca influx, and substantially reduced the proportion of cells that gave electrophysiological responses to either agent. Bradykinin also activated polyphosphoinositidase C in the dorsal root ganglion neurons, elevating levels of inositol(1,4,5)-trisphosphate and 1,2-diacylglycerol, an endogenous activator of PKC. It is suggested, therefore, that PKC may mediate some of the effects of bradykinin in sensory neurons. PMID- 2552044 TI - Supratentorial anaplastic gliomas in adults. The prognostic importance of extent of resection and prior low-grade glioma. AB - A retrospective analysis is presented of factors affecting the length of survival of 285 consecutive adults with newly diagnosed biopsy-proven supratentorial anaplastic glioma (188 cases of glioblastoma multiforme, 76 of anaplastic astrocytoma, 11 of anaplastic mixed glioma, and 10 of anaplastic oligodendroglioma) treated at a regional cancer center from July, 1982, through December, 1987. The approach to initial therapy included maximum feasible resection and radiotherapy. The median survival time for all patients was 35 weeks. Multivariate analysis demonstrated that age, duration of symptoms, preirradiation performance status, tumor histology, accessibility to resection, extent of resection, radiotherapy, and prior low-grade glioma were significant independent variables influencing survival. The prognostic importance of age, duration of symptoms, performance status, and tumor histology are already recognized, but three "new" findings are reported. First, patients with anaplastic oligodendroglioma had the longest median survival time (278 weeks). Second, corrected for accessibility and all other variables, patients with gross total resection lived longer than those with partial resection, and patients with any degree of resection lived longer than those who underwent only a biopsy procedure. Third, patients with anaplastic glioma in whom there was a prior history of low-grade glioma lived significantly longer after the diagnosis of anaplastic glioma than did patients in whom the anaplastic glioma apparently arose de novo. PMID- 2552045 TI - Repeat transsphenoidal surgery for Cushing's disease. AB - Transsphenoidal resection of adrenocorticotrophic hormone (ACTH)-producing pituitary adenomas has the potential of curing most patients with Cushing's disease. However, transsphenoidal exploration of the pituitary is not always curative, and patients who have remission of hypercortisolism following surgery occasionally develop a recurrence. Whether repeat pituitary surgery should be performed for recurrent or persistent Cushing's disease has not been evaluated previously. To determine the efficacy of transsphenoidal surgery in recurrent or persistent Cushing's disease, we performed transsphenoidal surgery in 31 patients (22 women and nine men) who had previously undergone a transsphenoidal operation and two female patients who had had previous pituitary irradiation only. In 24 (73%) of the 33 patients, remission of hypercortisolism was achieved by surgery. Although preoperative computerized tomography (CT) scanning identified an adenoma in only three of the 33 patients, in 20 patients a discrete adenoma was identified at pituitary exploration. The incidence of hypercortisolism was greatest if an adenoma was identified at surgery and the patient received selective adenomectomy (19, or 95% of 20 patients), if there was evidence at surgery or by preoperative CT scanning that the previous surgical exposure of the pituitary was incomplete (seven, or 78% of nine patients), if an adenoma was seen on preoperative CT scanning (three of three patients), or if the patient had had prior pituitary irradiation without surgery (two of two patients). In contrast, only five (42%) of 12 patients who received subtotal or total hypophysectomy had remission of hypercortisolim. Surgically induced hypopituitarism occurred in six (50%) of these 12 patients, but in only one (5%) of the 20 patients who underwent selective adenomectomy. Three (13%) of the 24 patients who were in remission from hypercortisolims following repeat surgery developed recurrent hypercortisolism 10 to 47 months postoperatively. Repeat transsphenoidal exploration of the pituitary and treatment limited to selective adenomectomy should be considered in patients with hypercortisolism despite previous pituitary treatment. If an adenoma is identified during surgery, the chance of remission of Cushing's disease is high and the risk of hypopituitarism is low; however, if no adenoma can be found and partial or complete hypophysectomy is performed, remission of hypercortisolism is less likely and the risk of hypopituitarism is about 50%. PMID- 2552046 TI - Characterization of insulin-like growth factor I and epidermal growth factor receptors in meningioma. AB - Receptors for insulin-like growth factor I (IGF-I) and epidermal growth factor (EGF) were localized and characterized in eight samples of human meningioma (four fibrous, two meningothelial, and two angioblastic types), using quantitative autoradiographic techniques. Effects of both growth factors on deoxyribonucleic acid (DNA) synthesis in the cultured meningioma cells were examined. High numbers of specific binding sites for both IGF-I and EGF were homogeneously present in tissue sections derived from fibrous and meningothelial types of meningiomas, whereas binding sites for these growth factors were not detectable in adjacent leptomeninges. While relatively large numbers of IGF-I binding sites were located in the wall of the intratumoral vasculature, the number of binding sites in the stromal component was lower in angioblastic-type meningiomas, including a low number of EGF binding sites detected only in the stromal portion. Scatchard analysis revealed the presence of a single class of high-affinity binding sites for both IGF-I and EGF in the meningiomas examined (dissociation constant (Kd) = 0.6 to 2.9 nM, and the maximum number of binding sites (Bmax) = 16 to 80 fmol/mg for IGF-I; and Kd = 0.6 to 4.0 nM, Bmax = 3 to 39 fmol/mg for EGF). Both growth factors increased the synthesis of DNA, in a dose-dependent manner, as measured by 3H-thymidine incorporation. The combination of IGF-I and EGF synergistically stimulated the synthesis of DNA, and the effects seen with 10% fetal bovine serum could be reproduced at a concentration of 10(-10) M. These observations can be interpreted to mean that both IGF-I and EGF may be involved in the growth modulation of meningiomas, possibly through paracrine or autocrine mechanisms. PMID- 2552047 TI - Attenuation of decompressive hypoperfusion and cerebral edema by superoxide dismutase. AB - This study tested the hypothesis that ischemia-reperfusion injury initiated by the superoxide anion radical is a major component of postdecompression hypoperfusion and cerebral edema, and could be attenuated by superoxide dismutase (SOD). A supratentorial extradural balloon was placed in 20 fasting, lightly anesthetized, mechanically ventilated dogs and inflated in 0.5-ml increments (0.07 ml/sec) at 15-minute intervals. The end-point of balloon expansion was the onset of an isoelectric electroencephalogram, near-arrest of hemispheric cerebral blood flow (CBF) (measured by H2 clearance), and the appearance of a suprainfratentorial intracranial pressure gradient, which was held for 15 minutes. The in vivo development of brain edema was detected by measuring brain elastic response (BER) extradurally, and was correlated with postmortem measurement of brain water content (gravimetry); blood-brain barrier integrity was tested by Evans blue dye given after the insult. After decompression, the dogs were randomly assigned to one of four treatment groups: Group I received hyperventilation (PaCO2 28 +/- 1 mm Hg, mean +/- standard deviation); Group II received furosemide (2.4 mg/kg) and pentobarbital (10 mg/kg) every 8 hours; Group III received 20% mannitol in a 1.4-gm/kg bolus plus furosemide, 0.5 mg/kg; and Group IV received SOD, 15,000 U/kg every 15 minutes for 3 hours. At 4 hours of decompression Group IV had significantly greater recovery in local CBF and BER than Groups I, II, and III (p less than 0.05). The 24-hour survival rate was 20% for Group I, 60% for Group II, 80% for Group III, and 100% for Group IV. The survival rate appeared to correlate with a variable degree of postmortem intraparenchymal hemorrhages, blood-brain barrier disruption, and moderate to severe brain edema for Groups I, II, and III. In contrast, Group IV had the least brain edema (p less than 0.05) and Evans blue dye extravasation (p less than 0.05) and the fewest intraparenchymal hemorrhages. These data support the hypothesis that, under the experimental conditions described here, the superoxide anion plays a major role in the pathophysiology of postdecompression ischemic edema. PMID- 2552048 TI - Radioimmunoimaging of lung vessels: an approach using indium-111-labeled monoclonal antibody to angiotensin-converting enzyme. AB - A murine monoclonal antibody against human angiotensin-converting enzyme was radiolabeled with 111In via diethylenetriaminepentaacetic acid without substantial loss of antigen-binding capacity. This monoclonal antibody designated 9B9 cross-reacted with rat and monkey angiotensin-converting enzyme. Indium-111 labeled 9B9 selectively accumulated 10-20 times greater in the lung than in blood or other organs following intravenous administration in rats. Kinetics of lung accumulation and blood clearance were studied for 111In-9B9-antibody and compared to that of 125I-labeled 9B9 in rat. Highly specific accumulation of 111In-9B9 antibody in the lung of Macaca Rhesus monkeys after intravenous injection was monitored by gamma-imaging. Images of 111In-labeled antibody 9B9 biodistribution in monkey lung noticeably differ from the images of biodistribution of 99mTc labeled albumin microspheres. This difference may provide information concerning the state of the endothelium of lung capillaries, which is different from the blood flow characteristics determined with routine microsphere technique. PMID- 2552049 TI - Focusing on learning. PMID- 2552050 TI - The response of higher education to the shortage of nursing school applicants. AB - This survey revealed that the majority of nursing schools responding, both public and private, required a 2.5 GPA for admission and that this standard had not been lowered in response to the nursing shortage. In general, responses indicated a belief that those students admitted with a GPA of less than 2.5 were not as likely to be successful in the program. Although some research in higher education suggests that more stringent requirements tend to improve the status of the college and therefore improve the quality of the applicants, the results of this survey did not support this. However, those schools that had increased their admission standards had done so within the past five years and, therefore, the effects of these changes may not yet be realized. The number, variety, and level of difficulty of prerequisite courses had remained constant and, in some cases, increased. Lowering the number and level of difficulty of prerequisite courses was not being done even with a decrease in the applicant pool. Required nursing courses were being offered on a more frequent basis by some schools to attract more students by increasing flexibility within the program and to increase the number of times during the year that students can be admitted. More time and money were being spent to improve the image of nursing and to recruit qualified students into the profession. Efforts were also being made to make it feasible for associate degree nurses to return for their bachelors degree in nursing.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552051 TI - Effect of a career decision-making course on the career planning of nursing students. AB - This study assessed whether participation in a structured career decision-making course would enhance the career planning of 42 final semester associate degree nursing students. Levels of career planning were measured by the Career Planning Scale (CPS) (Super, Thompson, Lindman, Jordaan, & Myers, 1982), and the Nursing Career Development Inventory (NCDI) (Savickas, 1984). Subjects were randomly assigned to either an experimental or control group. The experimental group received a four-week career decision-making course, which included self assessment of values, interests, and skills, and information on occupational opportunities in nursing. An ANOVA on the change scores from pretest to post-test revealed significance between the groups on the CPS (p = .002), and on the NCDI (p less than .00001). The experimental group progressed much more in career planning. All subjects were found to have lower levels of career planning than the average college student at pretest. Intercorrelational data and multiple regression analyses indicated that individuals who had worked in a nursing related field had higher levels of career planning. Overall, the course emerged as being effective in enhancing the career planning of nursing students. PMID- 2552052 TI - Lesbian phobia among BSN educators: a survey. AB - This exploratory study identified the attitudes of BSN educators in a midwestern state towards lesbians. A 50% (N100) return rate of mailed questionnaires elicited responses to 48 statements about lesbians and lesbianism on a modified Likert scale. The questionnaire included general demographic data and responses to statements seeking information about topics of lesbianism or lesbian issues. Specific areas addressed included moral and ethical responses, knowledge statements, sexuality or sex related statements, normalness of lesbians and lesbian behavior, and social contact with lesbians. Notable findings revealed that BSN educators were not personally fearful of sexual advances from lesbians; however, they did indicate concern about lesbians molesting and caring for children as well as lesbians in the role of educators. More than half believed that lesbianism is not a natural expression of human sexuality. PMID- 2552053 TI - The development of a master's degree program based on perceived future practice needs. AB - The need for an accessible master's degree program was determined through informal communication between practicing nurses and nursing educators. The purpose of the descriptive study was to project a master's degree program based on perceived future nursing practice needs. Forty-seven nurse administrators and staff nurses responded to a mailed survey (61.8%). The respondents were asked to make predictions regarding the probability of occurrence of 18 previously Delphi generated events by the year 2000. Respondents also provided judgments regarding the degree of agreement/disagreement that various content items would be critical to survival in practice and best learned in a master's degree program. Most respondents agreed that general education content items were both critical to survival and best learned by master's education. Less strong agreement that nursing content items were critical to survival and best learned in a master's degree program was an unexpected finding. A multidisciplinary master's program for all health-care professionals was proposed. PMID- 2552054 TI - Comparison of expected and evidenced baccalaureate degree competencies. AB - The defining of beginning competencies for the baccalaureate degree in nursing education has been an issue over the past decade. Many researchers have attempted to investigate the performance of graduates of the basic nursing preparation programs. At present, there is little that links education to practice. The purposes of this study were to validate whether the competencies of graduates of baccalaureate degree nursing programs were as expected by nursing service administrators, and whether the expected competencies were evidenced in the work setting. A two-part questionnaire was administered to 142 directors of nursing in health-care settings in Mississippi. The research project requested an evaluation of the baccalaureate competency statement from two perspectives. On a five-point Likert scale, administrators were asked to evaluate if the competency should be expected of a baccalaureate graduate. The administrators then evaluated to what extent the competency statement was evidenced. Analysis of the data was done using descriptive statistics. A mean for each competency was calculated for expected and evidenced responses. A difference of 30% between the expected and evidenced responses was deemed to be significant. Conclusions were drawn and recommendations for further study made. PMID- 2552055 TI - Nursing summer camp: a recruitment experience for high school students. PMID- 2552056 TI - Healthworks: a leadership experience. PMID- 2552058 TI - Grantsmanship as a core component of nursing doctoral education. PMID- 2552057 TI - Inter-university collaboration in the development of a distance education course in community health nursing. AB - Collaboration between faculty members from two university nursing programs has successfully produced an introductory community health nursing course for RN-BScN distance education students. The course has integrated theoretical and clinical components organized into a coherent whole via a detailed course manual. The course is designed to be flexible so that individual units of the course manual can be reorganized as needed to fit the curriculum requirements of the co authoring university. The process of collaboration was both frustrating and satisfying. The frustrations included unavoidable difficulties as well as problems that, now that they have been identified, could be avoided in future. The greatest satisfaction, of course, was that of actually achieving a worthwhile goal despite the obstacles that presented themselves during the course development process. PMID- 2552059 TI - Effects of neutral detergent fiber from blackgram (Phaseolus mungo) in rats and rabbits. AB - The hypocholesterolemic action of neutral detergent fiber (NDF) from blackgram (Phaseolus mungo) in rats may be due to hemicellulose, since removal of hemicellulose resulted in the loss of cholesterol-lowering action. Blackgram NDF showed different binding affinities for different bile acids, with maximum binding observed with chenodeoxycholic acid, and minimum binding with deoxycholic acid. Fecal excretion of inorganic cations (Mn++, Mg++, Cu++, Fe , Zn++, Na+ and K+) was greater in rats fed blackgram NDF than in rats fed a fiber-free diet. Absorption of glucose and cholesterol was lower in rabbits fed blackgram NDF than in rabbits fed a fiber-free diet. In vitro studies revealed significant binding of inorganic cations and bile acids by blackgram NDF. Cholic acid may be adsorbed on the surface of blackgram NDF. PMID- 2552060 TI - Impact of prospective payment and discharge location on the outcome of hip fracture. AB - OBJECTIVE: To determine the impact of prospective payment by diagnosis-related groups (DRGs) on length of stay in the hospital, ambulatory status, and level of post-hospital care needed for patients hospitalized with hip fracture. DESIGN: Retrospective chart review of a consecutive series of cases before and after the reference date of implementation of the prospective payment system (PPS). SETTING: Academic, tertiary-care hospital. PATIENTS/PARTICIPANTS: 181 patients 69 years of age or older admitted with International Classification of Diseases (ICD) or DRG codes for hip fracture. RESULTS: Length of stay was shorter by 1.37 days in the post-PPS era (p = 0.05). Poorer discharge ambulation was found in the post-PPS group (p = 0.089). At one year, differences in ambulation and nursing home residence were found to be related not to the implementation of PPS, but rather to the nursing home to which the patient was discharged. Patients discharged to a facility with active physical rehabilitation were less likely to remain institutionalized (p = 0.0025) than those in "ordinary" nursing homes and ambulated more independently (p = 0.05). CONCLUSIONS: The PPS did not have a significant long-term impact on hip fracture outcome. Post-hospital care may be of crucial importance to the future quality of life of hip fracture patients. PMID- 2552062 TI - Inhibition of tooth eruption through calcium-phosphate ceramic granules in the rat. AB - This experiment investigated the reason for disturbances of tooth eruption through hydroxylapatite (HA) granules. HA granules of compact and porous structure were implanted in the pathway of erupting teeth in 14 five-day-old rats. The results demonstrate that the use of this material can result in tooth retention and malformed tooth crowns. Tooth crown malformation can be traced to the formation of ceramo-dentinous complexes which promote bacterial invasion of the pulp cavity, resulting in arrest of the tooth eruption process. PMID- 2552061 TI - Sequential analysis of monomorphic and polymorphic major histocompatibility complex antigen expression in human heart allograft biopsy specimens. AB - Changes in major histocompatibility complex (MHC) antigen expression after heart transplantation were investigated in 233 cardiac allograft biopsy specimens of 33 patients by means of immunohistologic examination. The altered tissue expression was related to histopathologic and clinical diagnoses. A panel of monoclonal antibodies directed to monomorphic determinants was used for the analysis of MHC antigens, class I (human leukocyte antigens [HLA]-A, B, C, and beta 2 microglobulin) and class II (HLA-DR, HLA-DP, HLA-DQ). Donor and recipient MHC antigen expression (HLA-A and HLA-B) was studied by use of monoclonal antibodies directed to polymorphic epitopes. It was found in 57 of 78 rejection episodes that the induction of class I MHC antigens on the normally negative myocyte membranes was related to the rejection process. Usually the induction was focally associated with lymphocytic infiltrates but in severe rejection was generalized on all myocyte membranes. After effective rejection treatment the class I induction was reversed. Class II (HLA-DR) MHC antigens were induced on most vessel endothelia. During rejection MHC antigens HLA-DP and HLA-DQ also were coexpressed on the endothelia of a few vessels. Donor HLA-A and HLA-B antigens were expressed by endothelial and interstitial cells in comparable density but only in low amounts on myocyte membranes. Recipient interstitial cells infiltrated around vessels with time after transplantation. Most interstitial cells between myofibrils, however, remained those of the donor type until 1 year after transplantation. These results show that cardiac allografts undergo remarkable changes in the expression of MHC antigens during clinical complications after transplantation. Furthermore, the changes in alloantigen composition may influence the clinical course. PMID- 2552064 TI - Optical probes of intradiskal processes in rod photoreceptors. II: Light scattering study of ATP-dependent light reactions. AB - Rod outer segment (ROS) disks, either stacked or freely floating, respond to flash illumination to yield a specific, ATP-dependent, light-scattering signal AL. In broken ROS AL signals occur only when AD signals have preceded them. The degree to which the preceding AD signal has been completed determines the amplitude of the following AL signal. However, in freshly detached ROS from dark adapted frogs Al signals with maximal size can be obtained without pre-incubation with exogenous ATP. The energized state, which is restored in broken ROS with the help of ATP, appears to prevail in the living retina and must therefore be considered to be "physiological". AL signals require structurally intact disks. Neither peripheral ROS proteins nor connecting filaments between adjacent disks are necessary. Their structural origin is the same as that of the preceding AD signal, i.e. osmotic disk swelling. AL signals consist of a single slow kinetic component (half-life 10 s at room temperature) and multiphase fast kinetic component (70 ms). The slow phase corresponds to a light-stimulated resumption of ATPase activity (this has been dealt with in a previous paper) whereas the fast component reflects an immediate response of the energized disk to the metarhodopsin I to metarhodopsin II transition. The latter effect is the subject of this paper. A variety of experiments, using different ATPase inhibitors, ionophores and membrane-permeable salts, have been carried out; they are all consistent with notion that AL originates in the disk interior and probes the existence of a proton electrochemical potential difference delta mu (H+) across the disk membrane. A model is presented which can explain all given properties of AL satisfactorily. According to this model the photolysis of rhodopsin causes a proton release in the disk lumen. This, in turn, results in osmotic swelling of the disks, provided that the internal buffer sites have been (at least partially) titrated with protons prior to the flash. Such conditions, i.e. a low internal pH, are provided by the proton transport across the disk membrane, which presumably takes place during the course of the preceding AD signal. PMID- 2552063 TI - A new, simple and accurate method for evaluating masticatory ability. AB - A new method has been devised to measure masticatory ability. Pigment-coated granules were prepared, and 730 mg (730-740 granules) were sealed in a rubber capsule. This capsule was used as a test material and was chewed by the subject under standardized conditions. The masticatory ability, defined as the joules of work performed, was calculated based on the concentration of pigment leaked from the crushed granules during the process of mastication. The procedure is rapid, simple and reliable, and can be used for a large number of clinical samples and studies. The measurement error, defined as the coefficient of variation, was 2.1%. The intra-individual variation of measurements of twenty volunteers ranged from 3.6 to 16.9%. A series of tests in each subject was necessary to obtain a reliable value of masticatory ability. PMID- 2552065 TI - Generation of free radicals by photoexcitation of pheophorbide alpha, haematoporphyrin and protoporphyrin. AB - An investigation of the production of radical species by photoexcitation pheophorbide alpha, haematoporphyrin and protoporphyrin was performed. In an aqueous solution containing different amounts of ethanol, the superoxide radical was detected by the spin trapping technique. In addition, secondary radicals were observed. The generation of oxygen radicals was found to dominate in solutions with a low ethanol content. PMID- 2552066 TI - Family history of convulsions and use of pertussis vaccine. AB - To evaluate the risk of neurologic events after vaccination with diphtheria tetanus-pertussis (DTP) vaccine, we used data from the Centers for Disease Control Monitoring System for Adverse Events Following Immunization to compare the family history of convulsions in persons reporting neurologic events with that in persons reporting nonneurologic events; these events have an onset within 3 days of immunization with DTP vaccine, given either alone or with oral poliovirus vaccine. Persons reporting neurologic events were 6.4 times more likely to report a prior personal history of convulsions than those reporting nonneurologic events (95% confidence interval 4.7 to 8.8), and were 2.4 times more likely to report a history of convulsions in first-degree family members, that is, siblings or parents (95% confidence interval 1.7 to 3.4). Similar risks were noted for subgroup analyses controlling for type of event (febrile vs nonfebrile convulsion), age at immunization, source of report, number of previous doses of DTP vaccine, and day of onset. Because the Centers for Disease Control monitoring system receives reports on a nonrandom sample of all adverse events after immunization, selection bias could not be ruled out. On the basis of these data, we conclude that children with a family history of seizures are at increased risk of neurologic events, primarily febrile convulsions, after DTP vaccination. However, this increase in risk may reflect a nonspecific familial tendency for convulsions rather than a specific vaccine effect. Considering the rare occurrence of neurologic events after DTP vaccination, the generally benign outcome of febrile convulsions (which make up the majority of these neurologic events), and the possible increased risk of pertussis in the general population if the estimated 5% to 7% of persons with a first-degree family history of convulsions were exempted from pertussis vaccination, we further conclude that a history of convulsions in siblings or parents should not be a contraindication to pertussis vaccination. Special care in the prevention of postvaccination fever may be warranted in children with a family history of seizures. PMID- 2552067 TI - Effect of various graft materials with tetracycline in localized juvenile periodontitis. AB - Ten patients with bilateral, posterior osseous defects associated with localized juvenile periodontitis (LJP) completed the study. Following the initial therapy, osseous defects were surgically debrided and grafted with a 4:1 volume ratio combination of either Synthograft/tetracycline (b-TCP/TTC), Periograf/tetracycline (HA/TTC) or freeze-dried bone allograft/tetracycline (FDBA/TTC). Graft materials were selected randomly for each half mouth following defect debridement, with a different material used on the opposite side for that patient. Immediately following each surgery, patients were placed on doxycycline 100 mg/day for 10 days. Direct re-entry evaluation of 51 osseous defects demonstrated no significant differences among the graft materials regarding hard tissue or soft tissue changes, except for greater percent defect fill for HA/TTC compared to b-TCP/TTC. Significant decreases in defect depth and pocket depth were achieved with each graft material. No adverse reactions to the use of any of the graft materials in combination with local and systemic tetracycline were found. The results indicate all three graft materials used in conjunction with TTC are acceptable and beneficial for the treatment and repair of osseous defects associated with localized juvenile periodontitis. PMID- 2552068 TI - HIV-1 peptides induce a proliferative response in lymphocytes from infected persons. AB - In a comprehensive search for T-cell epitopes of HIV-1, several new regions were discovered. The analysis was performed with lymphocytes of HIV-1-infected persons in various stages of the infection. Peptides covering the entire group antigen (gag), and transmembrane (gp41) regions, and one-half of envelope (gp120) regions of HTLV-IIIB were studied. Both common and patient-unique responses were identified. Twelve common gag T-cell sites were discovered, as well as patient unique activating peptides. The gag peptides elicited the most frequent cell responses and the responses remained in late stages of disease. Only 1 of the 12 T-cell activating gag peptides was reactive with specific anti-HIV IgG. Eighteen common env-representing peptides evoked T-cell responses. They could be grouped into four previously undescribed regions of gp120 and two known sites, the hypervariable stretch and part of the CD4 binding region. Cell responses to env peptides were common in early stages of disease and tended to decrease in ARC and AIDS. The gp41-representing peptides evoked a cellular response to a region close to the N-terminus of the hydrophobic transmembrane region. In addition to the T cell activating peptides, peptides representing p15 and p19 as well as previously recognized regions of gp120 and gp41 appeared to be potent B-cell epitopes. PMID- 2552069 TI - Prevalence of HTLV-I compared to HIV-1 and HIV-2 antibodies in different groups in the Ivory Coast (West Africa) AB - Different Ivorian population groups were comparatively tested for antibodies to HTLV-1, HIV-1, and HIV-2. They included 1,334 healthy individuals, 176 unselected medical patients in regional hospitals, 149 female prostitutes, and 65 males with a sexually transmitted disease (STD) collected in 1987 from four regions (east, north, west, and central). The HTLV-1 prevalence averaged 1 to 2.7% in the different regions, without significant increase in sexually overexposed groups. Furthermore, in Dabou Protestant Hospital, 60 km west of Abidjan, 414 blood donors, 109 asymptomatic HIV-positive individuals, and 181 AIDS clinical cases were also tested. While a significant increase in the prevalence of HTLV-1 antibodies was observed only in AIDS patients. HIV-1 and HIV-2 prevalence increased, as expected, from a low rate (2.4%) in the general population to an intermediate rate (13-15%) in blood donors and unselected hospitalized patients and to a high rate (35%) in prostitutes and STD male patients. PMID- 2552070 TI - The antinociception produced by intrathecal morphine, calcium, A23187, U50,488H, [D-Ala2, N-Me-Phe4, Gly-ol]enkephalin and [D-Pen2, D-Pen5]enkephalin after intrathecal administration of calcitonin gene-related peptide in mice. AB - Calcitonin gene-related peptide (CGRP), which has been shown to modulate calcium in both the brain and in peripheral tissues, has not previously been shown to modulate calcium in the spinal cord. This study shows that the effects of CGRP given intrathecally (i.t.) appear to result from calcium modulation. Evidence for this hypothesis is the parallel shift to the right in the dose-effect curves of both i.t. calcium and i.t. A23187 (a calcium ionophore) by i.t. CGRP, and the enhancement by ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid of the ability of CGRP to antagonize morphine-induced antinociception. The CGRP induced modulation of spinal calcium appears to occur in opiate-sensitive pathways as well as nonopioid pathways, as evidenced by parallel shifts to the right in the dose-response curves of i.c.v. and i.t. morphine in either the tail flick or the hot-plate test and nonparallel shifts in the dose-response curve of s.c. morphine in the tail-flick test. CGRP attenuates the antinociceptive effect of the delta receptor-specific ligand [D-Pen2, D-Pen5]enkephalin (i.t.), but enhances the antinociceptive effect of the kappa ligand, U50, 488H. CGRP does not appear to interact directly with the opiate receptor because it does not mimic the activity of naloxone i.t. However, CGRP-induced alterations of calcium in opiate-sensitive spinal pathways appear to produce subtle modulation of opiate antinociception without direct opiate receptor interaction. PMID- 2552071 TI - Variation in sensitivity of six cat and six rat arteries to norepinephrine can be related to differences in agonist affinity and receptor reserve. AB - The sensitivity of contraction to norepinephrine and alpha-1-adrenoceptor affinity and reserve were measured in six rat and six cat arteries. These were the thoracic and abdominal aorta, superior mesenteric, renal, and femoral arteries of both species, and rat tail and cat splenic arteries. Sensitivity to norepinephrine differed by more than a factor of 30 in the rat and 20 in the cat. Rank order of sensitivities were in general similar in the two species. In rat and cat there was a significant correlation between sensitivity to norepinephrine and alpha-1 adrenoceptor affinity and also between sensitivity and receptor reserve, expressed as -antilog (pD2 - pKA). In the rat the contribution of affinity to these differences in sensitivity was greater than that of receptor reserve. In the cat arteries, receptor reserve is the more important factor. These results support the "variable receptor affinity hypothesis." This proposes that the affinity of a receptor can vary, and this may be due to differences in receptor structure, local membrane microenvironment, and extent of influence of intracellular mechanisms. The hypothesis proposes that variation in affinity can have a significant impact on tissue sensitivity. PMID- 2552072 TI - Chronic ethanol treatment selectively increases the binding of inverse agonists for benzodiazepine binding sites in cultured spinal cord neurons. AB - The effect of chronic ethanol treatment, and its withdrawal on the binding of ligands to the benzodiazepine binding sites of gamma-aminobutyric acid (GABA) receptor complex, was investigated in C57BL/6J mice spinal cord cultured neurons. Chronic ethanol (50 mM) treatment increased the specific binding of inverse agonists of the benzodiazepine binding sites, without affecting the binding of agonist or antagonist to this site. Thus, chronic ethanol exposure of the neurons increased the binding of [3H] Ro 15-4513 [ethyl-8-azido-5,6-dihydro-5-methyl-6 oxo-4H-imidazo [1,5-alpha][1,4]benzodiazepine-3-carboxylate] and methyl-1-beta carboline-3-carboxylate [( 3H]beta-CCM), but not the binding of [3H]flunitrazepam or [3H]Ro 15-1788 [ethyl-8-fluro-5-6-dihydro-5-methyl-6-OxO-4H-imidazo[1,5 alpha][1, 4] benzodiazepine-3-carboxylate]. This increase was due to an increase in the number of binding sites for Ro 15-4513 and beta-CCM, and not due to a change in receptor affinity. The increase was observed as early as after a 12-hr exposure of the neurons with ethanol, and remained elevated at 24-hr withdrawal, returning to control values at 48-hr withdrawal. These results further strengthen the notion that Ro 15-4513 and related inverse agonists binding site on the GABA benzodiazepine receptor complex may be involved in ethanol-s behavioral, biochemical and pharmacological effects which are mediated via GABAA receptor system. The significance of the enhanced binding sites for Ro 15-4513 and beta CCM in the actions of ethanol, tolerance and withdrawal is discussed. PMID- 2552073 TI - 1,3-Di(2-tolyl)guanidine blocks nicotinic response in guinea pig myenteric neurons. AB - Ditolylguanidine (DTG) is a ligand which binds with high affinity to neuronal sigma receptors. Activation of sigma receptors inhibits the release of acetylcholine (ACh) from guinea pig ileum myenteric plexus preparations. A study was therefore undertaken to investigate the action of sigma receptor ligands on single neurons. Nicotinic responses to locally applied ACh onto single neurons of the guinea pig ileum myenteric plexus were studied using intracellular recording techniques. DTG and (+)-SKF10047 (N-allylnormetazocine) produced a concentration dependent suppression of the depolarization of enteric neurons evoked by ionophoresis of ACh. The EC50 values for DTG and (+)-SKF10047 were 4.7 and 3.8 microM, respectively, and were similar to that for hexamethonium (3.2 microM). The inhibition of the ACh-depolarization was not mediated at sigma receptors because (-)SKF10047 and Bridge-DPG (2-imino-1,3H-dibenzo[d,f]-[1,3]-diazepine), which are inactive at sigma receptors, were as potent as DTG and (+)-SKF10047. DTG and hexamethonium (each at 1 microM) were more effective blockers of ACh induced inward currents at a holding potential of -100 mV than at -40 mV. This voltage dependence is consistent with a channel blocking mechanism. DTG (10 microM) did not affect the depolarization (mediated by 5-HT3 receptors) induced by pressure application of 5-HT onto single neurons. DTG and Bridge-DPG inhibited contractures of the longitudinal muscle-myenteric plexus preparation elicited by dimethylphenylpiperazinium noncompetitively (EC50 values were 8.0 and 12.3 microM, respectively) whereas DTG but not Bridge-DPG inhibited 5-HT-induced contractions of the longitudinal muscle-myenteric plexus noncompetitively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552074 TI - Role of low Km cyclic AMP phosphodiesterase inhibition in tracheal relaxation and bronchodilation in the guinea pig. AB - This study evaluated the relationship between inhibition of the rolipram sensitive and the CI-930-sensitive low Km cyclic AMP-specific phosphodiesterase (PDE) isozymes (PDE IIIRO and PDE IIIc, respectively) and bronchomotor tone in the guinea pig. Rolipram and CI-930 exhibited biphasic concentration-response relationships for relaxation of carbachol-, histamine- and leukotriene D4 contracted trachea. However, each agent produced a monophasic (sigmoidal) concentration-response curve when tested in the presence of a fixed concentration (3 microM) of the other. The same relationships were observed for inhibition of tracheal peak III PDE isolated via diethylaminoethyl-cellulose chromatography. Whereas CI-930 was approximately equipotent inhibiting PDE IIIc and relaxing rolipram-pretreated trachea, rolipram was substantially more potent (EC50 = 0.02 microM) in relaxing CI-930-pretreated trachea than in inhibiting CI-930 pretreated PDE III (PDE IIIRO, IC50 = 2.6 microM). Among a series of PDE inhibitors, there was a highly significant correlation (r = 0.89, P less than .01) between PDE IIIc inhibition (i.e., PDE III in the presence of rolipram) and rolipram-pretreated tracheal relaxation, but not between PDE IIIRO inhibition and CI-930-pretreated tracheal relaxation (r = 0.23). Nine of the PDE inhibitors used in this study have been reported to displace rolipram from a high-affinity binding site in rat brain. A highly significant correlation between relaxation of CI-930-pretreated trachea and displacement of rolipram binding by these agents was observed (r = 0.97, P less than .0001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552075 TI - Effect of chronic administration of U-50,488H on tolerance to its pharmacological actions and on multiple opioid receptors in rat brain regions and spinal cord. AB - The effects of chronic administration of U-50,488H (trans-3,4-dichloro-N-[2-(1 pyrrolidinyl)cyclohexyl]benzeacetamide ), a selective kappa opioid agonist, on the development of tolerance to its analgesic and hypothermic effects and on mu, delta and kappa opioid receptors in brain regions and spinal cord of male Sprague Dawley rats were determined. Rats were injected i.p. twice daily with 25 mg/kg of U-50,488H for 4 days. The development of tolerance to the analgesic and hypothermic effects of U-50,488H was almost complete after 4 days of treatment. [3H]Tyr-D-Ala-Gly-MePhe-Gly-ol (DAMGO), [3H]Tyr-D-Ser-Gly-Phe-Leu-Thr (DSTLE) and [3H]ethylketocyclazocine (EK) were used as ligands for mu, delta and kappa opioid receptors, respectively. The binding of [3H]DAMGO to membranes prepared from various brain regions (pons + medulla, midbrain, hypothalamus, corpus striatum and cortex) and spinal cord was unaffected by chronic administration of U 50,488H. The binding of [3H]DSTLE in U-50,488H-treated rats was decreased in spinal cord and increased in corpus striatum. The binding of [3H]EK to membranes prepared from pons + medulla, midbrain, cortex and spinal cord was decreased whereas it was increased in the corpus striatum. The changes in the binding of [3H]DSTLE and [3H]EK after chronic treatment with U-50,488H were due to changes in the maximum binding values and not in the Kd values. The results indicate that, in the rat, chronic administration of U-50,488H results in the development of tolerance to its analgesic and hypothermic effects and down-regulation of kappa and delta opioid receptors in the spinal cord and an up-regulation in the corpus striatum.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552076 TI - Role of renal nerves in excretory responses to administration of kappa agonists in conscious spontaneously hypertensive rats. AB - The present study examined whether the renal sympathetic nerves contribute to the renal excretory responses produced by kappa opioid receptor agonist administration in conscious spontaneously hypertensive rats (SHR). Intravenous infusion of the kappa opioid receptor agonists, ketocyclazocine (KC) and U 50488H, produced increases in urine flow rate. KC and U-50488H infusion also resulted in a marked and sustained antinatriuresis which was promptly reversed by low-dose naloxone (50 micrograms/kg i.v.), thus suggesting an opioid receptor mediated action of both agonists. Although these kappa agonists did not produce changes in glomerular filtration rate or renal plasma flow, efferent renal sympathetic nerve activity increased with the same time course as the antinatriuretic response. To investigate whether the decrease in urinary sodium excretion was mediated via the increase in efferent renal sympathetic nerve activity, experiments were repeated in SHR with prior bilateral renal denervation. These studies demonstrated that similar renal excretory responses (diuresis and a naloxone reversible antiinatriuresis occurred during infusion of KC and U-50488H in renal denervated as were seen in intact SHR. These studies indicate that the renal excretory responses to the kappa opioid agonists KC and U 50488H are not mediated through changes in renal hemodynamics or via a pathway requiring intact renal innervation. Because an antinatriuretic response was observed in renal denervated SHR, this suggests that kappa opioid receptor agonists may influence the renal tubular reabsorption of sodium by additional naloxone-sensitive mechanisms independent of intact renal innervation. PMID- 2552077 TI - Spinal analgesic actions of kappa receptor agonists, U-50488H and spiradoline (U 62066). AB - Administered i.p. to mice, the selective kappa receptor agonists U-50488H and spiradoline (U-62066) were more potent on the tail-flick than on the hot-plate analgesic assay. Both were more potent after i.s. rather than i.c. administration, a result consistent with earlier demonstrations that tail-flick analgesia is generally dependent upon spinal mechanisms. Intraspinal U-50488H was not effective in elevating rat tail-flick latencies. Both drugs increased the thresholds for cat spinal cord nociceptive neurons to respond to a noxious heat stimulus. However, maximal responses of spinal cord neurons to nociceptive stimuli were not altered. It is concluded that although spinal cord sites may be critical to kappa receptor analgesic mechanisms, the effects are quite distinct from spinal cord effects observed previously with classical narcotic analgesics. PMID- 2552078 TI - Nonlinear relationship between benzodiazepine receptor occupancy and glucose metabolic response in the conscious mouse brain in vivo. AB - To evaluate the relationship between the pharmacological effect of benzodiazepine (BZP) and BZP receptor (BZP-R) binding in the conscious mouse brain, a response of the local cerebral metabolic rate of glucose utilization (GU) to clonazepam (CNZ) was measured as an index for the pharmacological effect. Two glucose analogs (3-O-[3H]methylglucose and 2-[14C]deoxyglucose) method, originally presented by A. Gjedde was used for determination of GU. In the cerebral cortex, GU decreased to 70 to 80% at 60 min after i.v. administration of CNZ (0.005-1.0 mg/kg), but CNZ did not change the lumped constant, and this effect was diminished completely by the administration of a BZP antagonist, Ro-15-1788 (5 mg/kg). The maximum effect of CNZ on GU (about 30% decrease) was found at 0.1 mg/kg of CNZ, but increasing the dose to 1 mg/kg had very little additional effect. In vivo BZP-R occupancy was measured using [3H]-Ro-15-1788. Receptor occupancy increased from less than 10% at a dose of 0.005 mg/kg up to essentially 100% at doses of 1 mg/kg or greater. ID50 in dose-response curve of the receptor occupancy for CNZ and ED50 in that of decrease in GU were 0.3 and 0.007 mg/kg, respectively . A nonlinear and hyperbolic relationship was observed between the receptor occupancy and the response for the glucose metabolic rate, indicating that BZP exert the maximum glucose metabolic change at a low fractional receptor occupancy (30-40%). PMID- 2552079 TI - Regulation of the mesocorticolimbic dopamine system by glutamic acid receptor subtypes. AB - Glutamic acid and excitatory amino acids specific for the glutamate receptor subtypes were microinjected into the A10 region of the rat. Glutamate produced an increase in motor behavior that was antagonized by pretreatment with the dopamine D2 receptor antagonist, haloperidol. This motor stimulant effect was produced by kainate, but not by N-methyl-D-aspartate (NMDA) or quisqualic acid. By using in vivo dialysis it was found that dopamine release in the nucleus accumbens and locomotor activity were enhanced by glutamate injection into the A10 region. Whereas glutamate was found to increase the postmortem concentration of dopamine metabolites in the medial prefrontal cortex, nucleus accumbens and A10 region, NMDA selectively increased dopamine metabolism in the prefrontal cortex, and kainate produced increases in the nucleus accumbens and A10 region. When glutamate and the NMDA receptor antagonist, 3-[(+/-)-2-carboxypiperazine-4 yl)propyl-1-phosphonic acid (CPP) were coadministered, CPP selectively abolished the effect of glutamate on medial prefrontal cortical dopamine metabolites. A physiological role for the NMDA receptor modulation of A10 dopamine neurons was shown by intra-A10 pretreatment with CPP antagonism of mild footshock-induced increase in dopamine metabolites in the prefrontal cortex. These data argue that glutamate is a regulatory transmitter of A10 dopamine neurons, and that the NMDA receptor subtype modulates neurons projecting to the prefrontal cortex whereas the kainate subtype modulates mesoaccumbens neurons. PMID- 2552080 TI - Calcium-sensitive and insensitive transient outward current in rabbit ventricular myocytes. AB - 1. A suction pipette whole-cell voltage-clamp technique was used to record membrane currents and potentials of isolated ventricular myocytes from rabbit hearts. 2. Transient outward current (Ito) was activated by voltage steps positive to -20 mV, increasing in amplitude with further depolarization to reach a maximum around +70 mV. The current attained its peak within 10 ms and then it inactivated for 100-200 ms. 3. A large portion of Ito still remained after the calcium current (ICa) was blocked when depolarizing pulses were applied at a frequency of 0.1 Hz or less. Therefore, this current component is referred to as calcium-insensitive Ito or It. 4. It showed voltage- and time-dependent inactivation similar to that observed in Purkinje fibres and other cardiac preparations. 5. The reversal potential of It depended on external K+ concentration, [K+]o, with a slope of 32 mV per 10-fold change in the presence of a normal [Na+]o (143 mM), while the slope was 48 mV per 10-fold change in low [Na+]o (1.0 mM). 6. It was completely inhibited by 2-4 mM-4-aminopyridine. Ito in the presence of ICa was also partially blocked by 4-aminopyridine and the remainder was abolished by 5 mM-caffeine. 7. The calcium-insensitive and caffeine sensitive Ito differed in their decay rates as well as in their recovery time courses. The former was predominantly available at a slow pulsing rate, while the latter increased its amplitude with high-frequency depolarization. 8. The caffeine-sensitive Ito was inhibited by a blockade of ICa, by replacing Ca2+ with Sr2+, by external application of ryanodine and by internal application of EGTA. This indicates that the current is calcium-sensitive and is dependent on increased myoplasmic Ca2+ through Ca2+ influx via the sarcolemma and Ca2+ release from the sarcoplasmic reticulum. The current is therefore designated as IK, Ca. 9. The physiological functions of IK, Ca and It are indicated by their contribution to ventricular repolarization at fast and slow heart rates, respectively. PMID- 2552082 TI - Characterization of three types of potassium current in cultured neurones of rat supraoptic nucleus area. AB - 1. Whole-cell, voltage-clamp recordings were obtained from neurones of the supraoptic area of neonatal rats in dissociated cell culture. Recordings were made from neurones having the same morphology as those which were vasopressin or oxytocin immunoreactive. 2. Three types of voltage-activated K+ current were identified on the basis of their kinetics, voltage sensitivities, Ca2+ dependence and pharmacology. The currents corresponded to the delayed rectifier current (IK), the A-current (IA), and the Ca2+-dependent current (IK(Ca] described in other neurones. 3. IK had a threshold of -40 mV, a sigmoidal time course of activation, and was sustained during voltage steps lasting less than 300 ms. The underlying conductance was voltage dependent reaching a maximum at +30 mV (mean maximum conductance 4.09 nS). The activation time constant was also voltage dependent declining exponentially from 4.5 ms at -30 mV to 1.8 ms at +50 mV. 4. IA was transient, and was activated from holding potentials negative to -70 mV; the maximum conductance (mean 5.9 nS) underlying the current was obtained at +10 mV. The activation and inactivation time constants were voltage dependent: the activation time constant declined exponentially between -40 mV (2.2 ms) and +40 mV (0.65 ms). 5. IK and IA were attenuated by the K+ channel blockers tetraethylammonium (TEA) and 4-aminopyridine (4-AP). TEA blocked the conductance underlying IK but appeared to alter the kinetics of IA. In contrast, 4-AP blocked the conductance underlying IA and, to a lesser extent, IK. 6. IK and IA were activated independently of external Ca2+ and the voltage activation of Ca2+ channels since these currents were recorded in the presence of Co2+, a Ca2+ channel blocker. 7. IK(Ca) was recorded only when Ca2+ (2 mM) was present in the external medium. From a holding potential of -30 mV, IK(Ca) had a threshold of 20 mV, was maximal at about +20 mV and declined at more positive potentials. This current was sustained during voltage steps lasting 100 ms and was abolished by addition of Co2+ (2 mM) to the medium. 8. The possible roles of the three K+ currents in regulating the characteristic firing behaviour of supraoptic neurones previously recorded in vivo and in vitro are discussed. PMID- 2552081 TI - Potassium channels involved in the transduction mechanism of dopamine D2 receptors in rat lactotrophs. AB - 1. Radioactive rubidium (86Rb+) efflux was used to measure potassium (K+) permeability in a study designed to asses both the presence and the sensitivity to ions and drugs of the K+ channels in the plasma membrane of rat lactotrophs. 2. Rb+ efflux from Rb+-pre-loaded lactotrophs into nominally calcium-free solution containing 5 mM-K+ was linear from 1 to 60 s, with a calculated rate of about 0.1%/s. Raising K+ concentrations to depolarize the cells stimulated the Rb+ efflux (0.2%/s), which was already significant after 1 s of exposure of the cell to 100 mM-K+. This component of Rb+ efflux has been designated component V (sensitive to voltage and Ca2+ independent). 3. Addition of Ca2+ to 5 mM-K+ solution had no effect on resting Rb+ efflux (0.1%/s), but did further stimulate Rb+ efflux into K+-rich solutions. This component, which has been designated component C, was completely inhibited by 0.5 mM-cadmium. These data fit the view that the increase in intracellular Ca2+ concentration during depolarization opens certain (Ca2+-activated) K+ channels. 4. K+ efflux was differently affected by K+ channel blockers. Tetraethylammonium (TEA) inhibited both V and C components while 4-aminopyridine (4-AP) inhibited the component V without modifying the C component of Rb+ efflux. 5. Dopamine appears to affect both types of Rb+ efflux components. Dopamine increased the efflux of Rb+ in a nominally Ca2+-free medium containing 5 mM-K+ (component V). This effect was statistically significant 15 s after exposure of the cells to 10 nM-dopamine. Increasing the concentrations of K+ to gradually depolarize the cells enhanced the rate of increase of Rb+ efflux induced by dopamine, being evident in the initial 2-5 s of incubations. Dopamine also increased Rb+ efflux in a 5 mM-K+ solution containing 1 mM-Ca2+ (component C). This effect was rapid (2-5 s) and inhibited by 0.5 mM-cadmium. The combined action of dopamine on both component C and V caused the cells to be less sensitive to depolarizing concentrations of K+. The increase in Rb+ efflux and the enhancement of prolactin release induced by high concentrations of K+ were, indeed, prevented by exposure of the cells to 10 nM-dopamine. 6. The effects of dopamine on either component V or component C were pharmacologically characterized as D2 receptor mediated, being mimicked by selective D2 receptor agonists (quinpirole and RU 24213) and stereospecifically blocked by the D2 receptor antagonist sulpiride.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2552083 TI - The inactivating K+ current in GH3 pituitary cells and its modification by chemical reagents. AB - 1. Whole-cell and single-channel recording techniques were applied to the study of the permeability and gating of inactivating K+ channels from clonal pituitary cells. 2. The cation selectivity sequence (measured from reversal potentials) for the channels underlying the inactivating K+ current was Tl+ greater than K+ greater than Rb+ greater than NH4+. The conductance sequence (determined from current amplitudes) was K+ = Tl+ greater than Rb+ greater than NH4+. 3. The inactivating current (IK(i] which was blocked by 4-aminopyridine (4-AP), activated at voltages more positive than -40 mV and half-inactivated at that voltage. Inactivation proceeded as the sum of two exponentials with mean time constants of 21 and 82 ms. Deactivation followed a single-exponential time course. 4. Recovery from inactivation was slow, voltage dependent and multi exponential, taking more than 50 s near the cell's resting potential. 5. The magnitudes of outward current and of slope conductance increased as the concentration of external K+ was increased. 6. On-cell and outside-out membrane patches revealed minicurrents with gating and pharmacological properties identical to whole-cell currents. Single channels with inactivating characteristics, while rarely observed, had an average slope conductance of 6-8 pS. 7. Intracellular application of the disulphonic stilbene derivative, SITS, and the protein-modifying reagent, N-bromoacetamide (NBA), at concentrations of 0.2-1 mM for several tens of minutes dramatically slowed the decay (inactivation) of K+ currents and caused coincident increases in the magnitude of outward IK(i). 8. Extracellular application of NBA at much lower concentrations (1-100 microM) and much shorter exposure times (1-30 s) also slowed inactivation. This effect was reversible for brief applications at low doses, but became irreversible after longer exposures. 9. Both internal and external NBA shifted the steady-state inactivation-voltage relation by +10 mV and reduced inactivation at voltages more positive than 0 mV. 10. The efficacy of external NBA was independent of holding potential between -80 and 0 mV. 11. Potassium minicurrents and single channels recorded from on-cell membrane patches were not affected by application of NBA to the extrapatch membrane. In contrast, NBA reversibly slowed the decay, increased the magnitude of minicurrents and prolonged the open times of single K+ channels recorded from outside-out patches. The single-channel conductance was unchanged by NBA.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2552084 TI - Transient calcium current of retinal bipolar cells of the mouse. AB - 1. Isolated bipolar cells were obtained by enzymic (papain) dissociation of the adult mouse retina. The membrane voltage was clamped and the membrane currents were measured by the whole-cell version of the patch-clamp technique. Isolated bipolar cells and horizontal cells of the goldfish retina were also studied for comparison. 2. Hyperpolarization from the holding voltage, Vh, of -46 mV evoked a slowly activating, Cs+-sensitive, inward current (probably an h-current), and depolarization evoked a TEA- and Cs+-sensitive outward current (probably a combination of K+ currents). 3. Depolarization from a more negative Vh (e.g. -96 mV) evoked a transient inward current that had maximal amplitude between -40 and 20 mV. This current was identified as a Ca2+ current (ICa): its amplitude was increased with elevated [Ca2+]o and was decreased with reduced [Ca2+]o, and it was blocked by 4 mM-Co2+, but not by 5 microM-TTX. 4. Both the perikaryon and the axon terminal generated ICa with similar properties. 5. The plot of Ca2+ conductance (gCa) against membrane voltage (activation curve) was sigmoidal: in 10 mM [Ca2+]o, gCa increased for membrane voltages more positive than -65 mV, was half-maximal at about -25 mV, and reached saturation at about +30 mV. The plot of inactivation of gCa against membrane voltage was also sigmoidal: with 1 s conditioning depolarization in 10 mM [Ca2+]o, gCa decreased for membrane voltages more positive than -80 mV, was half-maximal at about -50 mV, and was fully suppressed for voltages greater than -30 mV. 6. ICa in the mouse bipolar cells was insensitive to 50 microM-Cd2+, 10 microM-nifedipine and 10 microM-Bay K 8644. In contrast, the calcium currents of bipolar and horizontal cells of the goldfish retina were markedly suppressed by 50 microM-Cd2+ and 10 microM-nifedipine, and were augmented several fold by 10 microM-Bay K 8644. The calcium currents of goldfish bipolar and horizontal cells were sustained, and were activated in a more positive range of potentials than the ICa of mouse bipolar cells. 7. The voltage range at which the ICa of mouse bipolar cells is activated includes the presumed range of membrane potentials spanned during light-evoked responses; thus, this current may participate in synaptic transmission. The transient character of ICa may also help to shape transient responses of ganglion cells. PMID- 2552085 TI - [Diagnosis of glucagonoma. Value of scanning, echography and arteriography. Apropos of 2 cases and a review of the literature]. AB - The authors report two cases of glucagonoma, a rare endocrine tumor of the pancreas, and describe the data currently found in literature. Glucagonoma is a single and usually large tumor, which develops in the alpha cells of the islets of Langerhans and evolves slowly. The combination of characteristic skin lesions, diabetes and weight loss should lead to searching for hyperglucagonemia and for the pancreatic tumor. The diagnosis is usually made rather late, average evolution is five years before diagnosis when it is detected. Imaging, in particular ultrasound and computed tomography (CT), proves to be necessary for the positive diagnosis of glucagonoma as it localizes the pancreatic mass and plays a role in local assessment, thus providing guidance for surgery. The role of imaging is also fundamental for the detection of metastases, which are the only sign of malignancy as no criterion of benignity is found for this tumor. PMID- 2552086 TI - Staff for general practice--short and long term options. PMID- 2552088 TI - Undergraduate departments of general practice: substance or symbolic shadow? PMID- 2552087 TI - International travel medicine. PMID- 2552089 TI - Symphysis-fundus measurements in screening for small-for-dates infants: a community based study in Gloucestershire. AB - Symphysis-fundus charts were introduced into 50 general practitioner antenatal clinics and the hospital antenatal clinics of two obstetricians in Gloucestershire in February 1985 for a 12-month period. Of the 1139 charts analysed, the sensitivity of one or more low fundal height measurements in predicting birthweight below the tenth centile for gestational age was 51% with a specificity of 88%. In the 319 charts with four or more measurements after 26 weeks gestation a sensitivity of 65% was recorded in predicting birthweight below the tenth centile, rising to 91% in the prediction of birthweight below the fifth centile; the specificity was correspondingly 81% and 80%. The sensitivity of the test varied inversely with maternal body mass index. The mean absolute difference in pairs of observations between general practitioners, midwives and an observer was 1.5 cm. Measurement of symphysis-fundus distance is not a precise diagnostic tool but it does provide an improvement on abdominal palpation in the prediction of small-for-dates infants. The findings of this study support the use of serial symphysis-fundus measurements in community antenatal clinics. Referral for ultrasound investigation is recommended when the measurement is low. PMID- 2552090 TI - Streptokinase used in general practice. AB - Over a six-month study in general practice 43 patients were identified whose presenting symptom was chest pain thought to be cardiac in origin. The median time from the onset of pain to the general practitioner attending was 60 minutes. On the basis of history, examination and initial electrocardiogram these patients were assessed as unlikely or likely to be infarcting. Of this latter group 15 fulfilled the inclusion criteria for intravenous streptokinase, four commencing treatment at home and 11 on admission to the local general practitioner medical ward. Each received 1.5 mega units over 60 minutes. The median time from the onset of pain to the start of therapy was 120 minutes. Of the 28 patients clinically suspected of having sustained a myocardial infarct 24 proved positive- an over-diagnosis rate of 14%. No major problems were encountered following streptokinase. PMID- 2552091 TI - Contact with general practitioners and differences in health status among people aged over 85 years. AB - A survey of the health and social service needs of all people aged 85 years and over was commissioned in 1986 by members of City and Hackney health authority and Hackney social services department. In 1987, 662 people who lived at home were traced from family practitioner committee records and interviewed. Eighty six per cent of respondents had consulted their general practitioner in the 12 months before the interview. Analyses showed that those who had not consulted within the 12 month period reported fewer physical and psychological symptoms, consumed less prescribed medication and reported better emotional well-being than those who had consulted more recently. However, the survey detected a considerable amount of unreported morbidity among both recent consulters and non-recent consulters. This finding questions the suggestion that the problem of under-consultation among elderly people is exaggerated but recommends that in studies of case finding attention should be focused on unreported morbidity rather than infrequent consultation. PMID- 2552092 TI - The family doctor and children with special educational needs. AB - Twenty three of the 1548 children registered with a group practice were found to have special educational needs. Only half of these children were known by the health visitor attached to the practice to have such needs. Ten of the 17 children attending school had no reference to their special needs in their case notes. Irregular attendance and poor immunization records indicated a lack of anticipatory care within the practice. Children with a physical handicap, in particular, attended their family doctor much less frequently than normal children or those with a mental disability. Improved care in recent years coincided with the establishment of developmental surveillance within the practice. PMID- 2552093 TI - Use of a contraindications checklist by practice nurses performing immunizations at a well child clinic. AB - At the inception of a general practice well child clinic, a checklist card was introduced into the clinic notes to summarize specific and relative contraindications to immunizations. This card was used by the practice nurses as they ran the immunization procedures during the clinic. A failure on the checklist led to a consultation with the clinic doctor who decided whether to proceed with the immunization. Of 155 immunizations given during the six-month period, only 23 (15%) failed the checklist and required the child to be assessed by the clinic doctor. Of these, nine (39%) were for simple upper respiratory tract infection. All the children were deemed fit to receive immunization. Only one child was found to have a specific contraindication to pertussis. The checklist cards allowed the smooth operation of the immunization procedures by practice nurses who were able to check comprehensively whether there were any contraindications and whether immunizations were being inappropriately refused. PMID- 2552094 TI - Modifying the behaviour of doctors and their receptionists in recurrent stressful situations. AB - Doctors and receptionists in a group practice completed a questionnaire about the image of the practice and reactions to recurrent stressful situations. The results were shared among the participants in a way that preserved anonymity. A year later a further questionnaire was completed which showed that respondents perceived that beneficial changes had taken place. The changes were seen as being mainly due to an increase in mutual understanding between the doctors and their receptionists. PMID- 2552095 TI - What influences doctors' prescribing? Sore throats revisited. AB - An audit of two practices in 1987 revealed a wide range of antibiotic prescribing for acute sore throat among the general practitioners. The data were presented at a postgraduate meeting and recommendations were made for a practice policy on antibiotic prescribing. The results of studies that looked at the objectives of treatment were included at that meeting. This paper presents a re-evaluation of the same doctors' antibiotic prescribing one-year later. Changes had occurred in the range and costs of drugs chosen, but individual doctors' prescribing rates remained broadly similar, in other words it was easier to influence what, but not whether, a doctor prescribes for this clinical condition. The existence of a prescribing 'threshold' within the individual doctor is supported. PMID- 2552096 TI - Communication between psychiatrists and general practitioners: what style of letters do psychiatrists prefer? AB - This study investigated the style of referral letter that psychiatrists would like to receive from general practitioners. Ninety psychiatrists in Edinburgh were asked to answer a brief questionnaire about their preferences and select one of six sample letters presented to them. The most popular letter was one page in length and contained two or three headings. PMID- 2552097 TI - Consultation rates in English general practice. AB - Methods of estimating the annual consulting rate per patient are reviewed. Methodological problems include the definition of consultations as opposed to problems encountered, the definition of population at risk, the reliability of data about home visits and the limitations of extrapolating data collected over a short period. Estimates of consultation rate are usually obtained from surveys which have other primary objectives. The annual consultation rate in 1981, excluding telephone contacts, was estimated at 3.5 consultations per patient. In spite of its limited sample size, the general household survey provides a reliable estimate of the national consulting rate. There is, however, a need to validate it against a survey covering a longer period in which consultation rates are measured and not just estimated from memory. The total workload of the 'average' doctor changed little between 1970 and 1981 in spite of reducing list size. Home visits accounted for approximately 15% of all consultations in 1981 and this value has been consistent over the period 1980-83. PMID- 2552098 TI - Out of the fire and into the frying pan: thoughts in anticipation of a new contract. AB - In anticipation of the government's review of the National Health Service an inner city doctor considers some of the issues raised by a contract which will challenge the general practitioner to manage a budget for both primary and secondary care and will require standards of care to be audited. There will be difficult clinical, management and ethical problems ahead, but these are capable of being solved. Such a contract holds out the prospect of enhanced health care for patients and a more fulfilled professional life for general practitioners. PMID- 2552099 TI - Oral contraceptives and intestinal ischaemia. PMID- 2552100 TI - Survey of analgesic use. PMID- 2552101 TI - The referral system and restrictive practices. PMID- 2552102 TI - Sterilizing instruments. PMID- 2552103 TI - Online information. PMID- 2552104 TI - AIDS and chastity. PMID- 2552107 TI - Mobility of injecting drug users. PMID- 2552106 TI - Heterosexual transmission of HIV. PMID- 2552105 TI - Children's immunization records. PMID- 2552108 TI - Vertical transmission of HIV infection. PMID- 2552109 TI - The participation of cyclic adenosine monophosphate (cAMP) in the regulation of meiotic maturation of oocytes in the laboratory mouse. PMID- 2552110 TI - Papillomavirus and vulvovaginal neoplasia. AB - Cumulative evidence strongly implicates human papillomavirus (HPV) in the genesis of squamous neoplasia of the lower female genital tract, including the vulva. The association of HPV with neoplasms at that site includes the relationship of specific HPV types with neoplasms and evidence that those HPV DNA types can transform epithelial cells in vitro. The capacity for in vitro transformation has been isolated to specific regions of the HPV genome. That may be unique in cancer associated viruses. Nevertheless, epidemiologic evidence points to additional factors, including immunologic, habitual and environmental, that may play an important role in the development of lower genital tract carcinomas. In particular, the marked differences in mean age and other variables between women with vulvar precancers and invasive cancer suggest that the evolution of invasive cancer involves more than HPV infection alone. Hence, the prevention of invasive vulvar cancer in older age groups will require an understanding of the unique host factors that render a small group of women susceptible to the disease in the face of an epidemic of HPV infection in the population at large. PMID- 2552111 TI - Hypothesis: possible role for the melatonin receptor in vitiligo: discussion paper. AB - A new unifying hypothesis for the aetiology of vitiligo is proposed, in which we postulate that the final destruction of melanocytes in vitiligo results from a cascade of reactions initiated by a disregulation of melanogenesis, caused by activation of the melatonin receptor. These events result in the high and uncontrolled production of free radicals and toxic products of melanogenesis which sequentially damage or destroy melanocytes and keratinocytes, provoke an autoimmune response against exposed intracellular or altered cell surface antigens, and increase the propensity of melanocytes to undergo malignant transformation. PMID- 2552112 TI - The glucagonoma syndrome. PMID- 2552113 TI - Dexamethasone-suppressible hyperaldosteronism: a large new kindred. AB - Dexamethasone-suppressible hyperaldosteronism (DSH) is a rare familial variety of primary aldosteronism in which the biochemical features of mineralocorticoid excess are corrected by treatment with glucocorticoids. We report a large new kindred with this syndrome, the second such family described in the British Isles and the first in Ireland. The family has a dramatic history of premature cardiovascular death. We have documented DSH in four members and found evidence of aldosterone excess in two others. The kindred is unique in that we have documented DSH in distant relatives (fourth cousins). All patients described are well controlled on potassium-sparing diuretics. Our report underlines the importance of a detailed family history in the assessment of hypertension due to primary aldosteronism. PMID- 2552115 TI - Novel benzodiazepine receptor partial agonists: oxadiazolylimidazobenzodiazepines. AB - The synthesis and biochemical evaluation of a series of oxadiazole derivatives of imidazobenzodiazepines related to the benzodiazepine antagonist Ro 15-1788 (2a) are reported. Although the oxadiazole ring is seen as an isosteric replacement for the ester linkage, significant differences in structure-activity trends were observed. Specifically, oxadiazoles 9-12 invariably had increased receptor efficacy (as witnessed by measurements of the GABA shift) relative to the corresponding ester. Additionally, and in direct contrast to the classical agonists such as diazepam, affinity for the benzodiazepine receptor was enhanced by a 7- rather than 8-halo substituent. The results are discussed in terms of a six-point receptor-binding model originally based on the X-ray structure of 2a. For comparison, the crystal structures of two representative oxadiazole derivatives, 10h and 12o, having a 6-oxo and 6-phenyl group, respectively, were determined and the data incorporated into a modified binding model to account for the greater efficacy of these compounds. It is concluded that the antagonist behavior of 2a relies upon the hydrogen-bond-acceptor properties of the ester carbonyl oxygen whereas for the oxadiazole series this site is localized at the imidazole nitrogen. PMID- 2552114 TI - Excitatory amino acid agonists. Enzymic resolution, X-ray structure, and enantioselective activities of (R)- and (S)-bromohomoibotenic acid. AB - The enantiomers of alpha-amino-4-bromo-3-hydroxy-5-isoxazolepropionic acid (4 bromohomoibotenic acid, Br-HIBO, 1) a selective and potent agonist at one class of the central (S)-glutamic acid receptors, were prepared with an enantiomeric excess higher than 98.8% via stereoselective enzymic hydrolysis of (RS)-alpha (acetylamino)-4-bromo-3-methoxy-5-isoxazolepropionic acid (4) using immobilized aminoacylase. The absolute configuration of the enantiomers of Br-HIBO was established by X-ray crystallographic analysis, which confirmed the expected preference of the enzyme for the S form of the substrate 4. (S)- and (RS)-Br-HIBO were potent neuroexcitants on cat spinal neurones in vivo, while (R)-Br-HIBO was a very weak excitant. Correspondingly, the S enantiomer of Br-HIBO (IC50 = 0.34 microM) was considerably more potent than the R form (IC50 = 32 microM) as an inhibitor of [3H]-(RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid ([ 3H]AMPA) binding to rat brain synaptic membranes in vitro. In contrast, (S)- and (R)-Br-HIBO were approximately equipotent (IC50 values of 0.22 and 0.15 microM, respectively) as inhibitors of [3H]-(S)-glutamic acid binding in the presence of CaCl2. The enantiomers of Br-HIBO showed no significant affinity for those binding sites on rat brain membranes which are labeled by [3H]kainic acid or [3H]-(R)-aspartic acid. PMID- 2552116 TI - Active reduced-size hexapeptide analogues of luteinizing hormone-releasing hormone. AB - A series of reduced-size hexapeptide analogues of LH-RH were synthesized that contain the residues corresponding to amino acid positions 4-9 and are linked to various carboxylic acids in place of residue 3. These compounds were tested in vitro in the rat pituitary receptor binding and LH release assays. A wide range of binding affinities was obtained up to and exceeding that of LH-RH. Both agonists and antagonists were obtained. From the SAR studies, it appears that a very precise size, length, and shape of the substituent at position 3 is required to achieve agonist activity, whereas the structural requirements for antagonist activity appear to be much less stringent. Depending on the nature of the substituent at positions 6 and 4, the biological response switches from antagonist to agonist or vice versa. The results suggest that conformational changes at position 6 or 4 feed back to the substituent at position 3, which induces the change from agonist to antagonist. The most potent compounds in the series were tested in vivo and found to be active. PMID- 2552117 TI - Tyrphostins I: synthesis and biological activity of protein tyrosine kinase inhibitors. AB - A novel class of low molecular weight protein tyrosine kinase inhibitors is described. These compounds constitute a systematic series of molecules with a progressive increase in affinity toward the substrate site of the EGF receptor kinase domain. These competitive inhibitors also effectively block the EGF dependent autophosphorylation of the receptor. The potent EGF receptor kinase blockers examined were found to competitively inhibit the homologous insulin receptor kinase at 10(2)-10(3) higher inhibitor concentrations in spite of the significant homology between these protein tyrosine kinases. These results demonstrate the ability to synthesize selective tyrosine kinase inhibitors. The most potent EGF receptor kinase inhibitors also inhibit the EGF-dependent proliferation of A431/clone 15 cells with little or no effect on EGF independent cell growth. These results demonstrate the potential use of protein tyrosine kinase inhibitors as selective antiproliferative agents for proliferative diseases caused by the hyperactivity of protein tyrosine kinases. We have suggested the name "tyrphostins" for this class of antiproliferative compounds which act as protein tyrosine kinase blockers. PMID- 2552118 TI - Novel linked antiviral and antitumor agents related to netropsin and distamycin: synthesis and biological evaluation. AB - A group of oligopeptides have been synthesized which are structurally related to the natural antiviral antitumor antibiotics netropsin and distamycin bearing two such moieties linked by polymethylene bridges. Cytostatic activity against both human and murine tumor cell lines and their in vitro activity against a range of viruses are reported. Enhanced antiviral activity was obtained against vaccinia virus. As a result of the introduction of the polymethylene linkers [( CH2)n, with n = 1, 2, and 6-8], both the antitumor and antivaccinia virus activity was markedly enhanced, relative to that of the parent compounds netropsin and distamycin. The biological activity of these agents is discussed both in terms of their structural differences and in relation to their minor groove binding to duplex DNA. PMID- 2552120 TI - Horizontal and vertical transmission of Japanese encephalitis virus by Aedes japonicus (Diptera: Culicidae). AB - Aedes japonicus (Theobald) was evaluated for competence as a vector of Japanese encephalitis virus and for its ability to transmit the virus vertically to its F1 larvae. Ae. japonicus supported the growth of the virus at 20 and 28 degrees C after feeding on a virus-blood mixture of 10(6.2) plaque forming unit (PFU)/ml. This species was able to transmit the virus to suckling mice after feeding on a virus-blood meal (10(6.2) PFU/ml) or a viremic chick (10(3.7) PFU/ml). Vertical transmission of the virus in Ae. japonicus was demonstrated with a minimum infection rate of 0.7%. PMID- 2552119 TI - Dihydropyrimidines: novel calcium antagonists with potent and long-lasting vasodilative and antihypertensive activity. AB - The novel calcium antagonists 3-N-substituted-3,4-dihydropyrimidines 1 and 9 and 3-N-substituted-dihydro-pyrimidin-2(1H)-ones 8 were regioselectively synthesized in good yields. Compounds 1 [especially 1s [R1 = (CH2)2N(benzyl)(2 naphthylmethyl), R2 = i-Pr, X = 0-NO2] and 1t [R1 = (CH2)2N(benzyl)(3,4 dichlorobenzyl), R2 = i-Pr, X = 0-NO2]] exhibited not only more potent and longer lasting vasodilative action but also a hypotensive activity with slow onset as compared with dihydropyridines. Moreover, some dihydropyrimidines [1q [R1 = (CH2)2N(benzyl)(3-phenylpropyl), R2 = CH2(cyclopropyl), X = 0-NO2], 1s, and 1t] were weaker in blocking atrioventricular conduction in anesthetized open-chest dogs and less toxic than the dihydropyridines. PMID- 2552121 TI - Larval diet and the vector competence of Culex annulirostris (Diptera: Culicidae) for Murray Valley encephalitis virus. AB - Culex annulirostris Skuse larvae were reared on two different amounts of powdered dog chow and yeast, low intake (2.8 micrograms/larva) and high intake (8.4 micrograms/larva), to produce adults with mean wing lengths of 3.25 and 3.7 mm, respectively. The resulting adults were fed different dosages of Murray Valley encephalitis virus and after 10 d extrinsic incubation at 28 degrees C, the infection rate, transmission rate, salivary gland, and body titers were determined. There were no significant differences in any of these parameters. PMID- 2552122 TI - Separation and properties of the haemolysins and extracellular enzymes of Listeria monocytogenes and L. ivanovii. AB - Desalted ammonium-sulphate (0-65%) precipitates from the cell-free supernates of 16-24-h cultures of Listeria monocytogenes Boldy and L. ivanovii (previously L. monocytogenes) Type 5 were eluted through Sephadex G-200. The enzyme activities gave rise to two main peaks. The first peak (approximate mol. wt of protein 150,000) contained only phosphatase activity (assayed by hydrolysis of 4 nitrophenylphosphate at pH 5.0 and 7.0). The second peak (approximate mol. wts of proteins 40,000-60,000) contained the haemolysin activity and the following hydrolytic activities (assay substrates are given in parentheses): phospholipase C (phosphatidyl choline and 4-nitrophenyl-phosphoryl-choline); phosphodiesterase (bis-4-nitrophenyl-phosphate); acid phosphatase (4-nitrophenylphosphatase); and esterases and lipases (4-nitrophenyl acetate, naphthyl-acetate and -oleate, triacetin and triolein). DEAE-Sephadex chromatography of appropriate fractions from the Sephadex G-200 purification step separated the first peak into two phosphatases and resolved the second peak into its constituent activities. Polyacrylamide gel electrophoresis showed that the individual fractions from the DEAE-Sephadex step consisted of mixtures of protein. The effects of pH and potential activators and inhibitors on the active proteins purified by DEAE Sephadex chromatography were examined. PMID- 2552123 TI - Volume-activated Na/H exchange activity in fetal and adult pig red cells: inhibition by cyclic AMP. AB - Hyposmotic swelling of pig red cells leads to a selective increase in K permeability, whereas hyperosmotic cell shrinkage augments the Na permeability. In this regard, the ouabain-resistant (OR) Na flux of red cells of newborn and adult pigs is characterized in detail. A reduction in cell volume by approximately 18% leads to an increase in the OR Na efflux of fetal and adult cells by 15- and fourfold, respectively. The OR Na influx in both cell types is equally influenced by cell shrinkage. Depletion of cellular K does not influence the volume-activated OR Na efflux. Nor does OR Na influx require external K. Both OR Na efflux and influx activated by shrinkage are inhibited by the diuretics furosemide and amiloride. The rank order of decreasing anion sensitivity for diuretic-sensitive Na efflux was acetate greater than chloride greater than gluconate greater than nitrate. Cell shrinkage induced by the addition of hypertonic salts results in an acidification of the unbuffered and CO2-free media, provided that both Na and DIDS are present. The acidification process can be reversed by either of the diuretic agents. These findings suggest that the shrinkage-activated OR Na flux is primarily mediated by a Na/H exchanger rather than by a Na/K/Cl cotransporter. Once loaded with either cAMP or cGMP, cell swelling can no longer activate the Na/H exchanger. The Na/H exchanger activity is detectable in the fetal cells of normal volume but quiescent in adult cells, indicating that the exchanger undergoes a developmental change during the transition from the fetal to adult stage. PMID- 2552124 TI - Effect of limited trypsin digestion on the renal Na+-H+ exchanger and its regulation by cAMP-dependent protein kinase. AB - The Na+-H+ exchanger from solubilized rabbit renal brush border membranes is inhibited by cAMP-dependent protein kinase (PKA) mediated protein phosphorylation. To characterize this inhibitory response and its sensitivity to limited proteolysis, the activity of the transporter was assayed after reconstitution of the proteins into artificial lipid vesicles. Limited trypsin digestion increased the basal rate of proton gradient-stimulated, amiloride inhibitable sodium uptake in reconstituted proteoliposomes and blocked the inhibitory response to PKA-mediated protein phosphorylation. To determine if the inhibitory response to PKA-mediated protein phosphorylation could be restored to the trypsin-treated solubilized proteins, nontrypsinized solubilized brush border membrane proteins were separated by column chromatography. The addition of small molecular weight polypeptides, fractionated on Superose-12 FPLC (Ve = 0.7), to trypsinized solubilized brush border membrane proteins restored the inhibitory response to PKA-mediated protein phosphorylation. Similarly, the addition of the 0.1 M NaCl fraction from an anion exchange column, Mono Q-FPLC, also restored the inhibitory response to PKA. Both protein fractions contained a common 42-43 kDa protein which was preferentially phosphorylated by PKA. These results indicate that limited trypsin digestion dissociates the activity of the renal Na+-H+ exchanger from its regulation by PKA. It is suggested that trypsin cleaves an inhibitory component of the transporter and that this component is the site of PKA-mediated regulation. Phosphoprotein analysis of fractions that restored PKA regulation raises the possibility that a polypeptide of 42-43 kDa is involved in the inhibition of the renal Na+-H+ exchanger by PKA-mediated protein phosphorylation. PMID- 2552125 TI - Ca2+ and cAMP activate K+ channels in the basolateral membrane of crypt cells isolated from rabbit distal colon. AB - Using patch-clamp techniques, we have studied Ca2+-activated K+ channels in the basolateral membrane of freshly isolated epithelial cells from rabbit distal colon. Epithelial cell clusters were obtained from distal colon by gentle mechanical disruption of isolated crypts. Gigaohm seals were obtained on the basolateral surface of the cell clusters. At the resting potential (approximately -45 mV), with NaCl Ringer's bathing the cell, the predominant channels had a conductance of 131 +/- 25 pS. Channel activity depended on voltage as depolarization of the membrane increased the open probability. In excised inside out patches, channels were found to be selective for K+ over Na+. Channel activity correlated directly with bath Ca2+ concentration in the excised patches. Channel currents were blocked by 5 mM TEA+ and 1 mM Ba2+. In cell-attached patches, after addition of the Ca2+ ionophore A23187, which increases intracellular Ca2+, open probability was markedly increased. Channel activity was also regulated by cAMP as addition of 1 mM dibutyryl-cAMP in the bath solution in cell-attached patches increased channel open probability over 20-fold. Channels that had been activated by cAMP were further activated by Ca2+. We conclude that the basolateral membrane of epithelial cells from descending colon contains a class of potassium channels, which are regulated by intracellular Ca2+ and cAMP. PMID- 2552126 TI - Activation kinetics of single high-threshold calcium channels in the membrane of sensory neurons from mouse embryos. AB - Activation kinetics of single high-threshold inactivating (HTI or N-type) calcium channels of cultured dorsal root ganglion cells from mouse embryos was studied using a patch-clamp method. Calcium channels displayed bursting activity. The open-time histogram was single exponential with an almost potential-independent mean open time tau op = 1.2 msec. The closed-time histogram was multicomponent; at least three of the components were associated with the activation process. The "fast" exponential component with the potential-independent time constant tau fcl = 0.9 msec included all intraburst gaps, while two "slower" ones with potential dependent time constants tau scl and tau vscl described shut times between bursts and between clusters of bursts. The burst length histogram was biexponential. The "fast" component with a relatively potential-independent time constant tau fbur = 0.6 msec described short, isolated channel openings while the "slow" component characterized real bursts with a potential-dependent mean life time. The waiting time histogram could be fitted by a difference of two exponentials with time constants being the same as tau scl and tau vscl. The data obtained were described in the frame of a 4-state sequential model of calcium channel activation, in which the first two stages are formally attributed to potential dependent transmembrane transfer of two charged gating particles accompanying the channel transitions between three closed states, and the third one to fast conformational changes in channel protein leading to the opening of the channel. The rate constants for all transitions were defined. The validity of the proposed model for both low-threshold inactivating (LTI or T-type) and high-threshold noninactivating (HTN or L-type) calcium channels is discussed. PMID- 2552128 TI - Formation of intermolecular and intramolecular hydrogen bonds in histidine binding protein J of Salmonella typhimurium upon binding L-histidine. A proton nuclear magnetic resonance study. AB - Histidine-binding protein J of Salmonella typhimurium has been chosen as a model system for a proton nuclear magnetic resonance spectroscopic investigation of binding protein-ligand interaction. This interaction is involved in the recognition step of the osmotic shock-sensitive active transport systems. When J protein binds L-histidine, four new, low-field, exchangeable proton resonances appear in the region +7 to +12 parts per million downfield from the water proton resonance (or +11.7 to +16.7 parts per million downfield from the methyl proton resonance of 2,2-dimethyl-2-silapentane-5-sulfonate). Due to their chemical shift range and other properties, they indicate the formation of both intra- and intermolecular hydrogen bonds. Experiments with 15N-labeled compounds confirm this conclusion. The specificity of the hydrogen-bond formation is demonstrated by observing the effects of substrate analogs, temperature, pH, and mutations on the exchangeable proton resonances. Proton-proton nuclear Overhauser effect measurements suggest that two of these exchangeable proton resonances (at +7.2 and +10.6 parts per million from H2O) are most likely from intramolecular hydrogen-bonded protons, while the other two (at +7.1 and +9.5 parts per million from H2O) are intermolecular hydrogen bonds. Our finding of L-histidine-induced hydrogen-bond formation in histidine-binding protein J in the solution state is an excellent demonstration of the production of specific conformational changes in a periplasmic binding protein upon binding of ligand. PMID- 2552127 TI - Conformational transitions and change translocation by the Na,K pump: comparison of optical and electrical transients elicited by ATP-concentration jumps. AB - The electrogenic properties of the Na,K-ATPase were studied by correlating transient electrical events in the pump molecule with conformational transitions elicited by an ATP-concentration jump. Flat membrane fragments containing a high density (approximately 8000 microm(-2)) of oriented Na,K-ATPase molecules were bound to a planar lipid bilayer acting as a capacitive electrode. ATP was released in the medium from a photolabile inactive ATP derivative ("caged" ATP) by a 40-microsec light flash. Electrical signals resulting from transient charge movements in the protein under single-turnover conditions were recorded in the external measuring circuit. In parallel experiments carried out under virtually identical conditions, the fluorescence of membrane fragments containing Na,K ATPase with covalently-bound 5-iodoacetamido-fluorescein (5-IAF) was monitored after the ATP-concentration jump. When the medium contained Na+, but no K+, the fluorescence of the 5-IAF-labeled protein decreases monotonously after release of ATP. In the experiments with membrane fragments bound to a planar bilayer, a transient pump current was observed which exhibited virtually the same time behavior as the fluorescence decay. This indicates that optical and electrical transients are governed by the same rate-limiting reaction step. Experiments with chymotrypsin-modified Na,K-ATPase suggest that both the fluorescence change as well as the charge movement are associated with the deocclusion of Na+ and release to the extracellular side. In experiments with Na+-free K+ media, a large inverse fluorescence change is observed after the ATP-concentration jump, but no charge translocation can be detected. This indicates that deocclusion of K+ is an electrically silent process. PMID- 2552130 TI - Phorbolester inhibits alpha 1-adrenoceptor mediated phosphoinositide breakdown in cardiomyocytes. AB - The regulation of and the intracellular events following alpha 1-adrenergic receptor stimulation of myocardium are not completely understood. The alpha 1 adrenergic stimulation of phosphoinositide breakdown was examined in a culture of neonatal rat ventricular myocytes and the influence of a protein kinase C activator, phorbol 12-myristate 13-acetate, on this process was studied. Inositolphosphate accumulation was stimulated by phenylephrine (EC50 5 microM) in the presence of 10 mM LiCl. The increase was antagonized by prazosin (10(-6) M) but not by propranolol (10(-6) M). The rate of inositolphosphate accumulation after prolonged alpha 1-adrenoceptor stimulation decreased without clear evidence of depletion of the membrane phosphatidylinositolbisphosphate pool. Phorbol ester treatment (IC50 10(-8) M) led to a dose-dependent inactivation of alpha 1 adrenoceptor stimulated phosphoinositide breakdown. These findings provide evidence that protein kinase C plays a role in the regulation of alpha 1 adrenoceptor sensitivity. PMID- 2552129 TI - Distribution and function of human ventricular beta adrenergic receptors in congestive heart failure. AB - This study examined the characteristics and distribution of sarcolemmal and light vesicular beta-adrenergic receptors (BAR) in left ventricular myocardium from 15 adults (aged 17 to 58 years) without left ventricular dysfunction or coronary artery disease and 29 patients (aged 14 to 53 years) with end-stage congestive heart failure (CHF). Sarcolemmal and intracellular fractions were prepared by 40,000 x g and 108,000 x g centrifugation, respectively. Agonist and antagonist binding properties were assessed by nonlinear computer modelling of isoproterenol 125I-pindolol (IPIN) displacement curves. Adenylate cyclase activity was also examined. Distribution of intracellular and sarcolemmal BAR was similar in normal and failing left ventricular myocardium, with intracellular BAR comprising 4.5 +/ 2.2% of total BAR in normal human heart and 5.7 +/- 5.1% of total BAR in CHF patients. For sarcolemmal BAR, antagonist affinity was similar for normal and CHF patients (KD IPIN in normals, 21.7 +/- 2.6 pM; KD IPIN in CHF, 20 +/- 2.3 pM). Agonist affinity was somewhat higher in CHF patients (KD isoproterenol in normals, 33 +/- 4.9 nM; KD isoproterenol in CHF, 6.2 +/- 1.5 nM). Sarcolemmal BAR number was reduced in CHF from 21.4 +/- 2.9 to 16.4 +/- 1.3 fmol/mg protein (P less than 0.04). Cyclic AMP production (pmol/mg protein/min above basal) was less in CHF after Gpp(NH)p stimulation (normals, 82 +/- 20; CHF, 27 +/- 9; P less than 0.01) and after stimulation with Gpp(NH)p + isoproterenol (normals, 129 +/- 25; CHF, 56 +/- 13; P less than 0.02). Stimulation with manganese + forskolin resulted in similar levels of cyclic AMP production in normals and in CHF patients. We conclude that: (a) sarcolemmal BAR number is reduced in CHF, but BAR are not redistributed intracellularly and (b) beta-adrenergic transmembrane signalling in CHF is also impaired at the level of the guanine nucleotide regulatory proteins. PMID- 2552132 TI - Genetics of N-methyl-N-nitrosourea induction of thymic lymphomas in AKR/J mice: assignment of a susceptibility gene to mouse chromosome 7. AB - Treatment of young AKR/J mice with N-methyl-N-nitrosourea (MNU) results in the induction of a high incidence of thymic lymphomas occurring between 4 and 6 months of age. The tumor incidence is higher and the latency period is shorter than that observed in other MNU-treated mouse strains. Analysis of tumor incidence in crosses of AKR/J with C57L/J mice indicates that several genes influence the incidence and latency of MNU-induced thymic lymphomas. One of these genes appears to be tightly linked to the albino locus of chromosome 7. PMID- 2552131 TI - Effect of sodium thiosulfate on the pharmacokinetics and toxicity of cisplatin. AB - Concurrent administration of sodium thiosulfate (STS) can protect against the nephrotoxic effects of even very-high-dose cisplatin (CDDP) (i.e., 270 mg/m2 given intraperitoneally). The effect of STS on the pharmacology and toxicity of CDDP was investigated in patients receiving at each treatment 90 mg of CDDP/m2 intraperitoneally, with STS given concurrently on alternate cycles by the intravenous route. The patients received a total of 38 courses of therapy, 21 without STS and 17 with STS. STS reduced the total exposure to diethyldithiocarbamate-reactive CDDP for the peritoneal cavity and plasma by 36% and 25%, respectively. When given alone, CDDP caused a statistically significant acute reduction in creatinine clearance levels; this reduction was less evident when STS was given. We conclude that, whereas STS does reduce systemic exposure, the magnitude of this effect was not sufficient to account for the ability of STS to protect against high-dose CDDP. PMID- 2552133 TI - Ultrastructural changes in rat Clara cells induced by bis(4-amino-3 methylcyclohexyl)methane. AB - Repeated oral administration of bis(4-amino-3-methylcyclohexyl)methane to rats induced unusual ultrastructural changes in Clara cells of the bronchiolar epithelium, involving marked accumulation of electron-dense inclusion bodies with a lamellar structure in the cytoplasm. The appearance of the inclusion bodies was speculated to result from accumulation of complexes formed between the agent and phospholipids within lysosomes, a condition known as drug-induced lipidosis. This finding appeared noteworthy, since it might reflect high potential for metabolism of xenobiotic compounds in Clara cells. PMID- 2552134 TI - A novel function of the herpes simplex virus type 1 Fc receptor: participation in bipolar bridging of antiviral immunoglobulin G. AB - We describe a novel function of the Fc receptor of herpes simplex virus type 1 (HSV-1), its ability to participate in antibody bipolar bridging. This refers to the binding of a single immunoglobulin G (IgG) molecule by its Fab end to its antigenic target and by its Fc end to an Fc receptor (FcR). We demonstrate that various immune IgG antibodies, including polyclonal rabbit antibodies to HSV-1 glycoproteins gC1 and gD1 and monoclonal human antibody to gD1 blocked rosetting of IgG-coated erythrocytes at IgG concentrations 100- to 2,000-fold lower than required for rosette inhibition with nonimmune IgG. Steric hindrance did not account for the observed differences between immune and nonimmune IgG since rabbit anti-gC1 F(ab')2 fragments did not block rosetting. Murine anti-gC1 or anti-gD1 IgG, a species of IgG incapable of binding by its Fc end to the HSV-1 FcR, also did not block rosetting. When cells were infected with a gC1-deficient mutant, anti-gC1 IgG inhibited rosetting to the same extent as nonimmune IgG. This indicates that binding by the Fab end of the IgG molecule was required for maximum inhibition of rosetting. Bipolar bridging was shown to occur even when small concentrations of immune IgG were present in physiologic concentrations of nonimmune IgG. The biologic relevance of antibody bipolar bridging was evaluated by comparing antibody- and complement-dependent virus neutralization of an FcR negative mutant and its parent HSV-1 strain. By engaging the Fc end of antiviral IgG, the parent strain resisted neutralization mediated by the classical complement pathway. These observations provide insight into the role of the HSV-1 FcR in pathogenesis and may help explain the function of FcR detected on other microorganisms. PMID- 2552135 TI - Persistent infection of rabbits with bovine leukemia virus associated with development of immune dysfunction. AB - Bovine leukemia virus (BLV) infection of rabbits provides a safe and relatively inexpensive in vivo mammalian system for the study of the mechanisms controlling expression of a unique group of lymphotropic retroviruses. This group of viruses, which includes C-type human T-lymphotropic virus types I and II and lentiviruslike human immunodeficiency virus type 1, possesses genes coding for "trans-activating" products. Rabbits experimentally inoculated with BLV became persistently infected, as demonstrated by a number of tests. All BLV-inoculated rabbits developed persistent serum antibody to BLV. Furthermore, all BLV inoculated rabbits had peripheral blood mononuclear cells which, when stimulated, expressed the virus, as demonstrated by viral induction of syncytium formation in a BLV-susceptible fibroblast line. The presence of BLV in circulating cells was confirmed by using peripheral blood mononuclear cells from randomly selected BLV inoculated rabbits, which showed the presence of viral reverse transcriptase activity, BLV transcriptional activity, or BLV proviral DNA. Additional tests showed that infected lymphocytes maintained in culture with recombinant human interleukin-2 formed multinucleated giant cells and produced virus when incubated in cytokine-containing medium. BLV-infected rabbits also showed alterations in several parameters associated with immunity, beginning 6 months after inoculation. Thirty-eight percent of infected rabbits developed abnormally low T cell responses, as measured by phytolectin stimulation, and T-cell responses cycled between normal and abnormally low over a period of 20 to 24 months. Forty four percent of rabbits infected for longer than 12 months suffered from recurrent conjunctivitis and rhinitis. By 24 months postinoculation, 28% of infected rabbits were dead or were killed because of poor clinical condition. PMID- 2552136 TI - Transforming activity of a 16-amino-acid segment of the bovine papillomavirus E5 protein linked to random sequences of hydrophobic amino acids. AB - The 44-amino-acid E5 protein of bovine papillomavirus type 1 is the smallest transforming protein yet described. Previous results from our laboratory indicate that a hydrophobic core and specific carboxyl-terminal amino acids are required for the E5 protein to exert its transforming function. In this study, additional substitution mutations were generated in the E5 gene to determine the minimal amino acid sequence requirements for focus formation in mouse C127 cells. In most cases examined, substitution of the hydrophobic middle third of the E5 protein with unrelated hydrophobic sequences severely inhibited transforming activity. However, we have identified one hydrophobic amino acid sequence apparently unrelated to the wild-type one that can replace the middle third of the wild-type E5 protein without affecting the ability of the protein to stably transform cells or interact with cell membranes. Furthermore, a mutant E5 protein in which only the carboxyl-terminal 16 amino acids of the protein have been derived from E5 sequences retains transforming activity. Since several residues in the carboxyl terminal portion of the E5 protein can be freely substituted with different amino acids (B. H. Horwitz, A. L. Burkhardt, R. Schlegel, and D. DiMaio, Mol. Cell. Biol. 8:4071-4078, 1988), the results reported here imply that much of the specific information necessary for cell transformation can be supplied by a subset of the carboxyl-terminal 16 amino acids of this protein. PMID- 2552137 TI - Respective roles of pyrimidine dimer and pyrimidine (6-4) pyrimidone photoproducts in UV mutagenesis of simian virus 40 DNA in mammalian cells. AB - UV light induces DNA lesions which are mutagenic in mammalian cells. We used simian virus 40 tsB201 (unable to produce viral capsid at the restrictive temperature of 41 degrees C because of a point mutation in the VP1 gene) to analyze the mutagenic potency of the two major UV-induced lesions, pyrimidine dimers (Py-Py) and pyrimidine (6-4) pyrimidones [Py(6-4)Py], which are formed on the same nucleotide sites. The mutagenesis criterion was the reversion toward a wild-type growth phenotype. After UV irradiation (mainly at 254 nm), part of the DNA was treated with the photoreactivating enzyme of Escherichia coli, which monomerizes Py-Py but does not modify the Py(6-4)Py photoproduct. Higher survival and lower mutation frequency rates for the photoreactivated DNA indicated that the two lesions were lethal and mutagenic. The VP1 gene of some mutants was entirely sequenced. The mutation spectra showed that the two lesions did not induce the same mutation hot spots, although some sites were common to both. The induced mutation hot spots were not only correlated with lesion hot spots but seemed partially directed by local DNA structures. PMID- 2552138 TI - Passive immunity modulates genetic reassortment between rotaviruses in mixedly infected mice. AB - Genetic reassortment between simian rotavirus SA11 and rhesus rotavirus (RRV) occurs with high frequency following mixed infection of nonimmune suckling mice (J. L. Gombold and R. F. Ramig, J. Virol. 57:110-116, 1986). We examined the effects of passively acquired homotypic or heterotypic immunity on reassortment in vivo. Passively immune suckling mice obtained from dams immune to either serotype 3 simian rotavirus (SA11) or serotype 6 bovine rotavirus (NCDV) were infected orally with either SA11 or RRV or a mixture of SA11 and RRV (both serotype 3 viruses). At various times postinfection, signs of disease were noted and the intestines of individual mice were removed and homogenized for titration of infectious virus and isolation of progeny plaques. Electrophoresis of genomic RNA was used to identify reassortants among the viral progeny isolated from infected animals. No reassortants (less than 0.45%) were detected among 224 clones examined from mixedly infected, homotypically immune mice. Twenty-nine reassortants (10.66%) were identified among 272 progeny clones from mixedly infected, heterotypically immune mice. Thus, reassortment was reduced more than 50-fold by homotypic immunity and approximately threefold by heterotypic immunity compared with prior data obtained from mixed infections of nonimmune mice. In addition, reassortment between SA11 and RRV in nonimmune mice was shown to be dependent on the virus dose. Taken together, these results suggest that immune responses may modulate the frequency of reassortment by reducing the effective multiplicity of infection (by neutralization or other immune mechanisms), thereby preventing efficient mixed infection of enterocytes. PMID- 2552139 TI - Receptor activity of rotavirus nonstructural glycoprotein NS28. AB - Rotavirus morphogenesis involves the budding of subviral particles through the rough endoplasmic reticulum (RER) membrane of infected cells. During this process, particles acquire the outer capsid proteins and a transient envelope. Previous immunocytochemical and biochemical studies have suggested that a rotavirus nonstructural glycoprotein, NS28, encoded by genome segment 10, is a transmembrane RER protein and that about 10,000 Mr of its carboxy terminus is exposed on the cytoplasmic side of the RER. We have used in vitro binding experiments to examine whether NS28 serves as a receptor that binds subviral particles and mediates the budding process. Specific binding was observed between purified simian rotavirus SA11 single-shelled particles and RER membranes from SA11-infected monkey kidney cells and from SA11 gene 10 baculovirus recombinant infected insect cells. Membranes from insect cells synthesizing VP1, VP4, NS53, VP6, VP7, or NS26 did not possess binding activity. Comparison of the binding of single-shelled particles to microsomes from infected monkey kidney cells and from insect cells indicated that a membrane-associated component(s) from SA11-infected monkey kidney cells interfered with binding. Direct evidence showing the interaction of NS28 and its nonglycosylated 20,000-Mr precursor expressed in rabbit reticulocyte lysates and single-shelled particles was obtained by cosedimentation of preformed receptor-ligand complexes through sucrose gradients. The domain on NS28 responsible for binding also was characterized. Reduced binding of single-shelled particles to membranes was seen with membranes treated with (i) a monoclonal antibody previously shown to interact with the C terminus of NS28, (ii) proteases known to cleave the C terminus of NS28, and (iii) the Enzymobead reagent. VP6 on single-shelled particles was suggested to interact with NS28 because (i) a monoclonal antibody to the subgroup I epitope on VP6 reduced particle binding, (ii) a purified polyclonal antiserum raised against recombinant baculovirus-produced VP6 reduced ligand binding, and (iii) a monoclonal antibody to a conserved epitope on VP6 augmented ligand binding. These experimental data provide support for the hypothesized receptor role of NS28 before the budding stage of rotavirus morphogenesis. PMID- 2552140 TI - Multiple isoelectric forms of poliovirus RNA-dependent RNA polymerase: evidence for phosphorylation. AB - Poliovirus-specific RNA-dependent RNA polymerase (3Dpol) was purified to apparent homogeneity. A single polypeptide of an apparent molecular weight of 63,000 catalyzes the synthesis of dimeric and monomeric RNA products in response to the poliovirion RNA template. Analysis of purified 3Dpol by two-dimensional electrophoresis showed multiple forms of 3Dpol, suggesting posttranslational modification of the protein in virus-infected cells. The two major forms of 3Dpol appear to have approximate pI values of 7.1 and 7.4. Incubation of purified 3Dpol with calf intestinal phosphatase resulted in almost complete disappearance of the pI 7.1 form and a concomitant increase in the intensity of the pI 7.4 form of 3Dpol. Addition of 32P-labeled Pi during infection of HeLa cells with poliovirus resulted in specific labeling of 3Dpol and 3CD, a viral protein which contains the entire 3Dpol sequence. Both 3Dpol and 3CD appear to be phosphorylated at serine residues. Ribosomal salt washes prepared from both mock- and poliovirus infected cells contain phosphatases capable of dephosphorylating quantitatively the phosphorylated form (pI 7.1) of 3Dpol. PMID- 2552141 TI - Beta and gamma interferons act synergistically to produce an antiviral state in cells resistant to both interferons individually. AB - We showed previously that the mouse fibroblastoid cell line Ltk-aprt- is resistant to the antiviral effects of beta interferon. This lack of response reflects a partial sensitivity to the interferon that is accompanied by a failure to activate expression of several interferon-regulated genes, although certain other genes respond in a normal manner. We show here that Ltk-aprt- cells were also unable to establish an antiviral state and to activate expression of 2,5 oligo(A) synthetase when treated with gamma interferon. Strikingly, however, treatment with a combination of beta interferon and gamma interferon provided complete protection against viral replication. Although the cells were completely insensitive to up to 250 U of the interferons per ml added singly, essentially complete protection from viral cytopathic effects was achieved when as little as 10 U of each of the interferons per ml were combined. Expression of 2,5-oligo(A) synthetase was also sensitive to this synergistic effect. Activation of an antiviral state could also be achieved by sequential treatment, first with gamma interferon and then with beta interferon. Partial protection against viral replication could be achieved by pretreatment with gamma interferon for as little as 1 h before incubation with beta interferon and could be blocked by the addition of specific antibodies or by cycloheximide, indicating that gamma interferon induces the synthesis of a protein which can act synergistically with a signal produced by the beta-interferon receptor. We suggest that Ltk-aprt- cells suffer from defects in one or more components of the gene activation pathways for both type I and type II interferons. Nonetheless, gamma interferon is able to activate the expression of a gene encoding a protein required for signal transduction. This protein acts synergistically with a transient signal produced in response to beta interferon, thereby activating the expression of a further group of genes. PMID- 2552142 TI - Herpes simplex virus type 1 ICP0 plays a critical role in the de novo synthesis of infectious virus following transfection of viral DNA. AB - As a first step in identifying the functions and intramolecular functional domains of herpes simplex virus type 1 infected cell protein 0 (ICP0) in productive infection and latency, a series of mutant plasmids specifying varying amounts of the ICP0 primary amino acid sequence were constructed. In transient expression assays with mutant and wild-type plasmids, the N-terminal half of the ICP0 molecule was found to be sufficient to transactivate a variety of viral promoters. Although promoters representing the immediate-early, early, and late kinetic classes were transactivated by wild-type ICP0, individual promoters responded to mutant forms of ICP0 in a manner consistent with the possibility that ICP0 transactivates different promoters by different mechanisms. Unlike infection with virus particles, which contain the 65-kilodalton transcriptional transactiovator, the initiation of viral replication after transfection of cells with purified viral DNA requires de novo protein synthesis. In order to assess the role of ICP0 in the de novo synthesis of infectious virus, Vero cells were transfected with purified DNA of wild-type virus or an ICP0 null mutant and the production of infectious virus was monitored. In cells transfected with mutant DNA, virus production was delayed by 2 days and the level of virus was reduced by several orders of magnitude relative to Vero cells transfected with wild-type viral DNA, suggesting an important role for ICP0 in the de novo synthesis of infectious particles. In cotransfection experiments with infectious DNA of the ICP0 null mutant and a plasmid specifying wild-type ICP0 titers of infectious virus were significantly enhanced relative to transfection with mutant DNA alone, confirming the role of ICP0 in de novo synthesis. These findings are consistent with the proposed role of ICP0 in reactivation of herpes simplex virus from latency (D. A. Leib, D. M. Coen, C. L. Bogard, K. A. Hicks, D. R. Yager, D. M. Knipe, K. L. Tyler, and P. A. Schaffer, J. Virol. 63:759-768, 1989), a process also thought to require de novo protein synthesis. The complementing activities of ICP0 mutant plasmids for ICP0 null mutant DNA in cotransfection assays correlated well with their transactivating activities for viral promoters in transient assays, indicating that the transactivating function of ICP0 is a critical factor in the de novo synthesis of infectious particles.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2552144 TI - Identification and characterization of an Epstein-Barr virus early antigen that is encoded by the NotI repeats. AB - The Epstein-Barr virus (EBV) genome is characterized by two regions carrying partially homologous clusters of short tandem repeats (NotI and PstI repeats) flanked by 1,044 and 1,045 base pairs with almost perfect homology (DL and DR, left and right duplications, respectively). Both repetitive regions are transcribed into poly(A)+ mRNA after induction of the productive EBV cycle with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate and contain open reading frames. To identify the potential protein encoded by the NotI repeat open reading frame (BHLF1), two repeat units of EBV strain M-ABA were expressed using the tryptophan-regulated Escherichia coli expression vector pATH11. Rabbit antisera generated against the resulting fusion protein reacted specifically with a protein varying in molecular size between 70,000 and 90,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, found after 12-O-tetradecanoyl phorbol-13-acetate or n-butyrate induction in various cell lines harboring EBV. In immunofluorescence tests with the BHLF1-specific antiserum, an immunofluorescence with EA-D specificity could be observed. In addition, the BHLF1 protein is exhibiting polyanion-binding activity with a maximum for single stranded DNA. Furthermore, the fusion protein is recognized by a number of human EBV-positive sera. PMID- 2552143 TI - The regions important for the activator and repressor functions of herpes simplex virus type 1 alpha protein ICP27 map to the C-terminal half of the molecule. AB - The herpes simplex virus type 1 (HSV-1) alpha or immediate-early proteins ICP4 (IE175), ICP0 (IE110), and ICP27 (IE63) are trans-acting proteins which affect HSV-1 gene expression. We previously showed that ICP27 in combination with ICP4 and ICP0 could act as a repressor or an activator in transfection assays, depending on the target gene (R. E. Sekulovich, K. Leary, and R. M. Sandri Goldin, J. Virol. 62:4510-4522, 1988). To investigate the regions of the ICP27 protein which specify these functions, we constructed a series of in-frame insertion and deletion mutants in the ICP27 gene. These mutants were analyzed in transient expression assays for the ability to repress or to activate two different target genes. The target plasmids used consisted of the promoter regions from the HSV-1 beta or early gene which encodes thymidine kinase and from the beta-gamma or leaky late gene. VP5, which encodes the major capsid protein, each fused to the chloramphenicol acetyltransferase gene. Our previous studies showed that induction of pTK-CAT expression by ICP4 and ICP0 was repressed by ICP27, whereas the stimulation of pVP5-CAT expression seen with ICP4 and ICP0 was significantly increased when ICP27 was also added. In this study, a series of transfection assays was performed with each of the ICP27 mutant plasmids in combination with plasmids containing the ICP4 and ICP0 genes with each target. The results of these experiments showed that mutants containing insertions or deletions in the region from amino acids 262 to 406 in the carboxy-terminal half of the protein were unable to stimulate expression of pVP5-CAT but were able to repress induction of pTK-CAT activity by ICP4 and ICP0. Mutants in the carboxy terminal 78 amino acids lost both activities; that is, these mutants did not show repression of pTK-CAT activity or stimulation of pVP5-CAT activity, whereas mutants in the hydrophilic amino-terminal half of ICP27 were able to perform both functions. These results show that the carboxy-terminal half of ICP27 is important for the activation and repression functions. Furthermore, the carboxy terminal 62 amino acids are required for the repressor activity, because mutants with this region intact were able to repress. Analysis of the DNA sequence showed that there are a number of cysteine and histidine residues encoded by this region which have some similarity to zinc finger metal-binding regions found in other eucaryotic regulatory proteins. These results suggest that the structural integrity of this region is important for the function of ICP27. PMID- 2552145 TI - Evidence for interference, coinfections, and intertypic virus enhancement of infection by ovine-caprine lentiviruses. AB - The ovine-caprine lentiviruses share nucleotide homology and serological properties in their gag-pol genes and gene products but constitute two distinct biological groups represented by ovine visna virus of Icelandic origin and by caprine arthritis-encephalitis and ovine progressive pneumonia viruses of U.S. origin. Two members of each group, visna 1514 and its antigenic variant LV1-1 in the first group and CAEV/CO and S93, a field isolate virus from a local arthritic sheep, in the second group, were examined in the present study in competitive binding studies in fibroblast and macrophage cell cultures. The cultures were preinoculated with each of the four viruses and then reinoculated with either 1514 virus or CAEV/CO, labeled with [35S]methionine. Both 1514 and CAEV/CO caused homologous interference. LV1-1 and S93 viruses shared the interference patterns of 1514 and CAEV/CO, respectively. 1514 and LV1-1 did not interfere with binding of CAEV/CO. Similarly, CAEV/CO and S93 did not interfere with binding of 1514. Remarkably, certain combinations, such as S93 plus 1514, resulted in enhanced binding of the second virus. Other experiments showed that the enhancement in binding extended to enhancement in replication of the second virus. These latter data suggested that individual cells supported replication of both viruses. Further testing of this phenomenon showed that goats could be doubly infected with two noninterfering viruses, 1514 and CAEV/CO. The ability of noninterfering related lentiviruses to infect the same cell and also the same host animal may be important in the natural history of these viruses in providing ideal conditions for the development of new recombinant viruses. PMID- 2552146 TI - Inhibition of rRNA synthesis by poliovirus: specific inactivation of transcription factors. AB - Synthesis of rRNA by RNA polymerase I is almost completely inhibited soon after infection of human cells with poliovirus. We show that extracts prepared from poliovirus-infected HeLa cells are severely inhibited in their ability to transcribe from a human rDNA promoter compared with extracts from mock-infected cells. Two lines of evidence presented here suggest that a specific transcriptional activity required for rDNA transcription in vitro is impaired in virus-infected cells. First, fractionation of individual transcriptional components by phosphocellulose chromatography and subsequent reconstitution experiments showed that the specific transcriptional activity of fraction C (0.8 M KCl eluate) from virus-infected cells was reduced three- to fourfold relative to that isolated from mock-infected cells. The activities of other transcription factors needed for in vitro transcription from the rDNA promoter were unaffected. Second, fraction C derived from mock-infected cells specifically restored transcription in extracts prepared from virus-infected cells. Fraction C contained both a nonspecific RNA polymerase I elongation activity and a specific factor activity which was needed for accurate transcription initiation. It is the specific transcriptional activity and not the nonspecific chain elongation activity of fraction C that is affected in cells infected with poliovirus. PMID- 2552147 TI - Use of Ar+ plasma etching to localize structural proteins in the capsid of herpes simplex virus type 1. AB - Partially cored herpes simplex virus type 1 (HSV-1) capsids (B capsids) were eroded in a low-energy (0.5-keV) Ar+ ion plasma under conditions in which the outermost structural proteins were expected to be degraded before more internal ones. After various periods of etching, the proteins remaining intact were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and determined quantitatively by densitometric scanning of the stained gels. The results showed that the major capsid polypeptide (VP5) and two other capsid proteins, VP19 and VP23, were degraded rapidly beginning as soon as capsids were exposed to the ion plasma. In contrast, significant lags were observed for erosion of VP21, VP22a, and VP24, suggesting that these proteins were available to accelerated ions only after other, more external structures had been damaged or eroded away. The results suggest that VP5, VP19, and VP23 are exposed on the surface of the capsid, while VP21, VP22a, and VP24 are found inside the capsid cavity. The experiments are consistent with the view that VP5 constitutes the major structural component of the hexavalent capsomers. It is proposed that VP19 and VP23 may form other surface structures such as the pentavalent capsomers, the capsid floor, or the intercapsomeric fibers. PMID- 2552149 TI - Degradation of cellular proteins during poliovirus infection: studies by two dimensional gel electrophoresis. AB - Picornaviruses encode for their own proteinases, which are responsible for the proteolytic processing of the polyprotein encoded in the viral genome to produce the mature viral polypeptides. The two poliovirus proteinases, known as proteins 2A and 3C, use the poliovirus-encoded polyprotein as a substrate. The possibility that these poliovirus proteinases also degrade cellular proteins remains largely unexplored. High-resolution two-dimensional gel electrophoresis indicates that a few cellular proteins disappear after poliovirus infection. Thus, at least nine acidic and five basic cellular proteins, ranging in Mr from 120 to 30 kilodaltons, are clearly degraded during poliovirus infection of HeLa cells. The degradation of these cellular polypeptides is very specific because it does not occur upon infection of HeLa cells with encephalomyocarditis virus or Semliki Forest virus. Moreover, inhibitors of poliovirus replication, such as cycloheximide or 3-methylquercetin, block the disappearance of these polypeptides. These results suggest that the input virions are not responsible for this degradation and that active poliovirus replication is required for the proteolysis to occur. Analysis of the time course of the disappearance of these polypeptides indicates that it does not occur during the first 2 h of infection, clearly suggesting that this phenomenon is not linked to the poliovirus-induced shutoff of host protein synthesis. This conclusion is strengthened by the finding that 3-methylquercetin blocks proteolysis without preventing shutoff of host translation. PMID- 2552148 TI - A host cell protein binds to a highly conserved sequence element (pac-2) within the cytomegalovirus a sequence. AB - The human cytomegalovirus (CMV) a sequence has significant homology to two regions, pac-1 and pac-2, within the a sequence of herpes simplex virus type 1 (HSV-1). Both regions have been shown to be important cis-acting signals in HSV-1 genome maturation. We have demonstrated that a small fragment from within the CMV a sequence, containing the pac-1 and pac-2 motifs, carries all of the signals necessary for generation of genomic termini and for inversion. These observations indicated that the function of these highly conserved sequence motifs was similar in CMV and HSV-1. We have identified and partially purified a host cell protein with affinity for the sequence 5'-GGCGGCGGCGCATAAAA-3' within CMV pac-2. This partially purified protein has an apparent molecular weight of 89,000 under denaturing conditions and could be renatured after sodium dodecyl sulfate polyacrylamide gel electrophoresis, suggesting that the capacity to bind DNA was the property of a single polypeptide chain. This activity was found in a wide variety of human cell lines, including those that are permissive as well as those that are nonpermissive for CMV growth, but not in cell lines from monkey, mouse, or drosophila origins. Our work implicates a host cell protein in a sequence function. PMID- 2552150 TI - Antigenic properties and cellular localization of herpes simplex virus glycoprotein H synthesized in a mammalian cell expression system. AB - Herpes simplex virus type 1 glycoprotein H (HSV-1 gH) was synthesized in an inducible mammalian cell expression system, and its properties were examined. The gH coding sequence, together with the stable 5' untranslated leader sequence from xenopus beta-globin, was placed under control of the strong promoter from the human cytomegalovirus major immediate-early gene in an amplifiable plasmid which contains the simian virus 40 (SV40) virus origin for replication (ori). This expression vector was transfected into ts COS cells constitutively expressing a temperature-sensitive SV40 T antigen which allows utilization of the SV40 ori at permissive temperatures. The results of transient expression assays at the permissive temperature showed that HSV-1 gH could be synthesized in greater amounts than those produced by a high-multiplicity virus infection. The proteins produced were detected in Western blots (immunoblots) with a HSV-1 gH-specific polyclonal serum raised against a TrpE-gH fusion protein. The transfected gH had an apparent molecular weight of approximately 105,000, intermediate in size to those of the precursor (100,000) and fully processed forms (110,000) of HSV-1 gH from infections. Antigenicity was investigated by reactions with three virus neutralizing monoclonal antibodies specific for conformational epitopes on gH. Only one of these monoclonal antibodies could immunoprecipitate the synthesized gH. However, equal recognition of the transfected gH was achieved by superinfection with virus. In addition, detectable amounts of gH were not expressed on the cell surface unless the cells were superinfected with virus. Studies with a temperature-sensitive mutant, ts1201, defective in encapsidation showed that the changes in antigenic structure and cell surface expression caused by superinfection with virus were not due simply to incorporation of gH into virions. These results suggest that gH requires additional virus gene products for cell surface localization and formation of an antigenic structure important for its function in mediating infectivity. PMID- 2552151 TI - Selective spread of herpes simplex virus in the central nervous system after ocular inoculation. AB - The spread of herpes simplex virus (HSV) was studied in the mouse central nervous system (CNS) after ocular inoculation. Sites of active viral replication in the CNS were identified by autoradiographic localization of neuronal uptake of tritiated thymidine. Labeled neurons were first noted in the CNS at 4 days postinoculation in the Edinger-Westphal nucleus, ipsilateral spinal trigeminal nucleus, pars caudalis, pars interpolaris, and ipsilateral dorsal horn of the rostral cervical spinal cord. By 5 days postinoculation, additional sites of labeling included the seventh nerve nucleus, nucleus locus coeruleus, and the nuclei raphe magnus and raphe pallidus. None of these sites are contiguous to nuclei infected at 4 days, but all are synaptically related to these nuclei. By 7 days postinoculation, no new foci of labeled cells were noted in the brain stem, but labeled neurons were noted in the amygdala, hippocampus, and somatosensory cortex. Neurons in both the amygdala and hippocampus receive axonal projections from the locus coeruleus. On the basis of these findings, we conclude that the spread of HSV in the CNS after intracameral inoculation is not diffuse but is restricted to a small number of noncontiguous foci in the brain stem and cortex which become infected in a sequential fashion. Since these regions are synaptically related, the principal route of the spread of HSV in the CNS after ocular infection appears to be along axons, presumably via axonal transport rather than by local spread. PMID- 2552154 TI - Protection of mice from herpes simplex virus-induced retinitis by in vitro activated immune cells. AB - A form of acute retinal necrosis occurred in the contralateral eyes of susceptible mice 1 week after each received a uniocular injection of live herpes simplex virus type 1 (HSV-1) in the anterior chamber. Although these mice did not develop systemic delayed hypersensitivity to virus antigens, their sera contained virus-specific antibodies at the time contralateral retinitis occurred. These findings suggest that systemic immunity might not be able to protect against contralateral retinitis. To explore this possibility further, we examined lymph nodes and spleens of intraocularly infected mice to determine whether their lymphoid tissues contained primed HSV-1-specific cytotoxic T cells. Virus specific cytotoxic T cells were readily identified in these mice. We wondered why successful immune priming did not confer protection against HSV-1 retinitis. We examined this issue by evaluating the capacity of in vitro-generated, HSV-1 specific effector T cells to prevent retinitis by infusing these cells by various routes and at various times into mice that received an intracameral injection of HSV-1. The results revealed that virus-specific effector cells could prevent contralateral retinitis if injected intravenously or into the anterior chamber of the contralateral eye at the same time that virus was injected into one eye. However, the effector cells failed to prevent retinitis if they were injected into the same eye that received HSV-1 or if their intravenous administration was delayed until 24 h after the HSV-1 injection into the eye. We concluded that immune T cells can protect against contralateral retinal necrosis caused by uniocular injection of HSV-1 into the anterior chamber but only if they are administered during the first 24 h after virus infection. We propose that a retinitis-inducing process is set in motion during this early time interval postinfection. Once the process has been initiated and established, it is no longer susceptible to immune intervention. It would appear that mice that are susceptible to contralateral retinitis fail to mobilize a protective response quickly enough to ward off the establishment of the retinitis-inducing process and its disastrous eventuality. PMID- 2552153 TI - A mutation in the short 5'-proximal open reading frame on Rous sarcoma virus RNA alters virus production. AB - The 5'-proximal open reading frame on Rous sarcoma virus RNA encodes a seven amino-acid peptide and is conserved in all avian sarcoma-leukosis retroviruses. Ribosome-binding site analysis in intact chick cells showed that the 5'-proximal AUG codon is a strong site for initiation of translation in vivo. Removal of the 5'-proximal AUG codon by site-specific mutagenesis resulted in a virus with a reduced ability either to replicate or to transform a population of chicken embryo fibroblasts. These results establish a procedure for determining sites of ribosome binding and initiation of translation on mRNAs in intact eucaryotic cells and strongly suggest that the 5'-proximal open reading frame (or its AUG codon) on Rous sarcoma virus RNA has an important role in regulating viral activity. PMID- 2552152 TI - Linker insertion mutants of simian virus 40 large T antigen that show trans dominant interference with wild-type large T antigen map to multiple sites within the T-antigen gene. AB - Linker insertion mutants affecting the simian virus 40 (SV40) large tumor (T) antigen were constructed by inserting a 12-base-pair oligonucleotide linker into restriction endonuclease cleavage sites located within the early region of SV40. One mutant, with the insertion at amino acid 5, was viable in CV-1p and BSC-1 cells, indicating that sequences very close to the amino terminus of large T could be altered without affecting the lytic infection cycle of SV40. All other mutants affecting large T were not viable. In complementation assays between the linker insertion mutants and either a late-gene mutant, dlBC865, or a host range/helper function (hr/hf) mutant, dlA2475, delayed complementation was seen with the 6 of the 10 nonviable mutants. Of these 10 mutants, 5 formed plaques 3 to 4 days later than in control complementations, while complementation by one of the mutants, inA2827, with an insertion at amino acid 520, was delayed more than 1 week. Most mutants which showed delayed complementation replicated less well in Cos-1 cells than did a control mutant, dlA1209, which produced no T antigen. The replication of inA2827(aa520) was reduced by more than 90%. Similar interference with viral DNA replication was seen when CV-1, HeLa, or 293 cells were cotransfected with an origin-defective plasmid encoding wild-type large T antigen and with inA2827(aa520). Only one of the mutant T antigens, inA2807(aa303), was unstable. These results indicate that some of the mutant T antigens interfered with functions of wild-type T required for viral DNA replication. However, not all of the mutants which showed delayed complementation also showed interference with viral DNA replication. This indicates that mutant large T antigens may interfere trans dominantly with multiple activities of wild-type large T antigen. PMID- 2552155 TI - Requirement of the single base insertion at the 3' end of the env-related gene of Friend spleen focus-forming virus for pathogenic activity and its effect on localization of the glycoprotein product (gp55). AB - In order to obtain evidence for the essential role of the single base insertion occurring at the 3' end of the env-related gene of Friend spleen focus-forming virus (SFFV) encoding the leukemogenic glycoprotein (gp55) a mutant SFFV genome was constructed in which the segment of the gp55 gene of the polycythemia inducing strain of SFFV containing the single base insertion and the 6-base-pair duplication was replaced by the corresponding sequence of the Friend murine leukemia virus env gene. The mutant SFFV-Friend murine leukemia virus complex did not induce symptoms of the erythroproliferative disease in adult DBA/2 mice. During passage through newborn DBA/2 mice, the mutant virus complex invariably gave rise to weakly pathogenic variant SFFVs. All of the variant SFFVs induced in adult DBA/2 mice a transient mild splenomegaly associated with normal or slightly low hematocrit value, and they produced gp55 with a molecular weight similar to that of gp55 of the wild-type SFFV. For the two isolates of variant SFFV, the 3' portion of the viral DNA intermediate containing the 3' portion of the gp55 gene was molecularly cloned. Nucleotide sequences of these biologically active cloned DNAs were determined and showed that the variant SFFV genomes arose from the mutant SFFV genome by regaining the single base insertion, indicating that the single base insertion is essential for the biological activity of gp55. Evidence is presented indicating that the single base insertion which causes a loss of the cytoplasmic domain of the env-related protein is not related to the localization of the further-glycosylated form of gp55 in the plasma membrane but is involved with the release of gp55 from cells. PMID- 2552156 TI - The herpes simplex virus virion host shutoff function. AB - The virion host shutoff (vhs) function of herpes simplex virus (HSV) limits the expression of genes in the infected cells by destabilizing both host and viral mRNAs. vhs function mutants have been isolated which are defective in their ability to degrade host mRNA. Furthermore, the half-life of viral mRNAs is significantly longer in cells infected with the vhs-1 mutant virus than in cells infected with the wild-type (wt) virus. Recent data have shown that the vhs-1 mutation resides within the open reading frame UL41. We have analyzed the shutoff of host protein synthesis in cells infected with a mixture of the wt HSV-1 (KOS) and the vhs-1 mutant virus. The results of these experiments revealed that (i) the wt virus shutoff activity requires a threshold level of input virions per cell and (ii) the mutant vhs-1 virus protein can irreversibly block the wt virus shutoff activity. These results are consistent with a stoichiometric model in which the wt vhs protein interacts with a cellular factor which controls the half life of cell mRNA. This wt virus interaction results in the destabilization of both host and viral mRNAs. In contrast, the mutant vhs function interacts with the cellular factor irreversibly, resulting in the increased half-life of both host and viral mRNAs. PMID- 2552157 TI - Derivation and characterization of an efficiently myocarditic reovirus variant. AB - A reovirus variant, 8B, was isolated from a neonatal mouse which had been inoculated with a mixture of two reovirus strains: type 1 Lang (T1L) and type 3 Dearing (T3D) (E. A. Wenske, S.J. Chanock, L. Krata, and B. N. Fields, J. Virol. 56:613-616, 1985). 8B is a reassortant containing eight gene segments derived from the T1L parent and two gene segments derived from the T3D parent. Upon infection of neonatal mice, 8B produced a generalized infection characteristic of many reoviruses, but it also efficiently induced numerous macroscopic external cardiac lesions, unlike either of its parents. Microscopic examination of hearts from infected mice revealed myocarditis with necrotic myocytes and both polymorphonuclear and mononuclear cellular infiltration. Electron microscopy revealed viral arrays in necrotic myocytes and dystrophic calcification accompanying late lesions. Determination of viral titers in hearts from T1L-, T3D , or 8B-infected mice indicated that growth was not the primary determinant of myocardial necrosis. Results from inoculations of athymic mice demonstrated that T cells were not a requirement for the 8B-induced myocarditis. Finally, 8B was more cytopathic than either of the parent viruses in cultured mouse L cells. Together, the data suggest that 8B-induced myocardial necrosis is due to a direct effect of reovirus on myocytes. Reovirus thus provides a useful model for the study of viral myocarditis. PMID- 2552158 TI - The reovirus M1 gene, encoding a viral core protein, is associated with the myocarditic phenotype of a reovirus variant. AB - Reoviruses contain a genome composed of 10 double-stranded RNA gene segments. A reovirus reassortant, 8B, derived from type 1 Lang (T1L) and type 3 Dearing (T3D), displayed a phenotype unlike that of either of its parents in that it efficiently induced numerous macroscopic external cardiac lesions in neonatal mice (B. Sherry, F. J. Schoen, E. Wenske, and B. N. Fields, J. Virol. 63:4840 4849, 1989). A panel of T1L/T3D reassortants and a panel of reassortants derived from 8B were used to determine whether novel T1L/T3D gene associations in 8B were responsible for its myocarditic phenotype. The results eliminated the possibility that any T1L/T3D gene combination found in 8B, from 2 genes to all 10 genes, was the explanation for its induction of cardiac lesions. This suggested that a mutation(s) in an 8B gene(s) might be responsible for induction of the myocarditis. Statistical analysis of experiments with 31 reassortants derived from 8B revealed a highly significant association (P = 0.002) of the 8B M1 gene with induction of cardiac lesions. The reovirus M1 gene encodes a viral core protein of unknown function, although evidence suggests a potential role in core structure and/or viral RNA synthesis. This represents the first report of the association of a viral gene with induction of myocarditis. PMID- 2552159 TI - Effects of mutations in poliovirus 3Dpol on RNA polymerase activity and on polyprotein cleavage. AB - A series of short insertion mutations was introduced into the poliovirus gene for 3Dpol at a number of different locations. When substituted for wild-type sequences in a full-length, infectious cDNA and tested for infectivity, all 3D mutants were nonviable. The mutant cDNAs were introduced into a bacterial plasmid designed to direct the expression of poliovirus 3CD, a viral protein composed of contiguous protease and RNA polymerase sequences. Bacteria transformed with these plasmids all expressed similar amounts of 3CD, and all mutant proteins cleaved themselves to generate wild-type 3Cpro and mutant 3Dpol polypeptides with approximately the same efficiency as wild-type 3CD. The released mutant 3Dpol proteins were all defective in RNA-dependent RNA polymerase activity in vitro. Uncleaved 3CD is a protease required for processing the viral capsid protein precursor, P1. In an in vitro assay of P1 cleavage activity, some of the mutant 3CD proteins expressed in Escherichia coli showed normal activity, while others were clearly inactive. Thus, alterations in the sequence and/or folding of different regions of the 3D protein have differential effects on its various activities. PMID- 2552160 TI - Spatial and temporal distribution of bovine herpesvirus 1 transcripts. AB - Northern (RNA) blot analysis was used to determine the spatial and temporal distribution of bovine herpesvirus 1 (BHV-1) transcripts. Total RNA was isolated from Madin-Darby bovine kidney cells which had been infected with BHV-1.2b strain K22 or BHV-1.1 strain Jura in the presence or absence of metabolic inhibitors. Cloned restriction fragments representing the entire genome of strain K22 were labeled with 32P and hybridized to immobilized RNA. A total of 54 BHV-1 transcripts were found, ranging in size from 0.4 to larger than 8 kilobases (kb). The inverted repeat regions and an adjacent segment of the unique large part of the BHV-1 genome encoded three major immediate-early (IE) transcripts and one minor IE transcript enriched after cycloheximide treatment of infected cells. Late transcripts were identified by drastically reduced abundance after cytosine arabinoside (araC) treatment. Twelve late transcripts were encoded mainly by the unique long genome region, with a cluster of four transcripts located on HindIII fragment K (map units 0.677 to 0.733). The 21 transcripts unaffected by araC treatment were defined as early; they showed dispersed locations over the whole genome, with a cluster on the unique short sequence. The 17 remaining transcripts could not be classified unambiguously as early or late by these techniques. The IE transcript with a size of 4.2 kb exhibited homology with the single IE gene of pseudorabies virus, and the IE transcript with a size of 2.9 kb was encoded in part by the genome region known to be transcriptionally active during latency. PMID- 2552161 TI - Polyomavirus genome and polyomavirus enhancer-driven gene expression during myogenesis. AB - The mRNAs for myogenic functions are coordinately transcribed with polyomavirus (Py) early mRNA during in vitro differentiation of mouse C2 myoblast cells. Sequence analysis shows that the A domain of the Py enhancer includes an E1A-like consensus sequence that is also found in the 5' upstream region of two genes expressed during myoblast differentiation: alpha-actin and myosin light chain. Therefore, the coordinate expression of such genes with Py early mRNA may be activated by a common cellular regulatory factor. In the present work, we report that C2 cells surviving Py infection are unable to differentiate and do not express alpha-actin and myosin light-chain mRNAs. Hybrids between such Py resistant myoblast cells and the parental cells exhibited dominance of the permissibility to Py growth and of the expression of myogenic mRNAs. In C2 cells transiently transfected with a chimeric plasmid (pSVPy12CAT) harboring the bacterial chloramphenicol acetyltransferase (CAT) gene driven by the Py enhancer promoter region, the CAT gene was expressed irrespective of their stage of differentiation. Moreover, undifferentiated stably transfected cells expressing the CAT gene restricted viral growth. Py-resistant C2 myoblasts transiently transfected with pSVPy12CAT also expressed the CAT gene driven by the Py enhancer. This contradictory finding is similar to results previously obtained by other investigators with cloned genes specific for myogenic functions, and it may be explained by a structural difference between the pSVPy12CAT and the Py genomic organizations in which the viral enhancer operates. PMID- 2552163 TI - Human papillomavirus type 27: detection of a novel human papillomavirus in common warts of a renal transplant recipient. AB - The cloning and partial characterization of the genome of human papillomavirus type 27 (HPV-27) is described. Hybridization analyses reveal that this is a new HPV type, with the strongest homology to HPV-2. PMID- 2552162 TI - Heterogeneity of the human papillomavirus group. PMID- 2552165 TI - Human papillomavirus type 29 (HPV-29), an HPV type cross-hybridizing with HPV-2 and with HPV-3-related types. AB - The cloning and partial characterization of human papillomavirus (HPV) type 29 is presented. By hybridization analyses, this virus appears to be related to HPV types associated with common warts and HPV types associated with flat warts. PMID- 2552164 TI - Human papillomavirus type 28 (HPV-28), an HPV-3-related type associated with skin warts. AB - The cloning and partial characterization of the genome of human papillomavirus type 28 (HPV-28) is presented. The virus is a distinct type and by hybridization analyses is most closely related to HPV-3 and HPV-10. PMID- 2552166 TI - Human papillomavirus type 48. AB - The cloning and partial characterization of the genome of human papillomavirus (HPV) type 48 is presented. Hybridization and short DNA sequence analyses permitted the alignment of the genome to the HPV genetic map. PMID- 2552167 TI - Human papillomavirus type 49, a type isolated from flat warts of renal transplant patients. AB - The cloning and characterization of the genome of human papillomavirus type 49 (HPV-49) is described. The viral DNA, which is most closely related to the DNAs of HPVs seen in patients with epidermodysplasia verruciformis, was aligned to the HPV-5 genome by electron microscopic analysis of heteroduplexes between the cloned viral DNAs. PMID- 2552168 TI - Human papillomavirus (HPV) type 50, a type associated with epidermodysplasia verruciformis (EV) and only weakly related to other EV-specific HPVs. AB - The cloning and partial characterization of the genome of human papillomavirus type 50 is presented. Alignment of the genomic map with that of human papillomavirus type 5, with which it is only weakly related, was permitted by partial DNA sequence analysis. PMID- 2552169 TI - Human papillomavirus type 53. AB - The cloning and partial characterization of the genome of human papillomavirus type 53 is presented. The virus is a distinct type and is most closely related to human papillomavirus type 30. PMID- 2552170 TI - In vitro mutagenesis of the herpes simplex virus type 1 DNA polymerase gene results in altered drug sensitivity of the enzyme. AB - A mutation (asparagine 815 to serine 815) was introduced into the herpes simplex virus type 1 (HSV-1) DNA polymerase (pol). The HSV-1 pol enzyme in lysates of Saccharomyces cerevisiae cells expressing the mutant protein showed increased resistance to acyclovir triphosphate and increased sensitivity to phosphonoacetate but was not substantially altered with respect to sensitivity to phosphonoformate or aphidicolin. These results directly demonstrate that both resistance to acyclovir triphosphate and sensitivity to phosphonoacetate can be conferred by this mutation in the absence of other viral factors and that the yeast expression system can be used for structure-function studies on HSV-1 pol. PMID- 2552171 TI - Analysis of regulatory elements of the equine infectious anemia virus and caprine arthritis-encephalitis virus long terminal repeats. AB - We analyzed the equine infectious anemia virus (EIAV) long terminal repeat (LTR) for sequences that influence its promoter activity and ability to be trans activated by the EIAV tat gene product. A series of LTR deletion mutants and recombinants between LTR and simian virus 40 (SV40) regulatory sequences were used for these studies. We were able to identify the EIAV promoter region and showed that sequences within the U3 region significantly inhibited LTR-directed transcription. However, when placed in a heterologous context (SV40 promoter) these U3 sequences functioned as an enhancer. trans-activation of the EIAV LTR was found to depend upon sequences downstream of the transcription initiation site and also within U3. Deletion mutagenesis experiments showed that the major downstream element was present in a 46-nucleotide stretch (+4 to +50). An SV40 promoter construct containing these sequences could be trans-activated in cells expressing the EIAV tat gene product. For comparative purposes we also examined the LTR of another animal lentivirus, caprine arthritis-encephalitis virus (CAEV), for positive and negative transcriptional regulatory elements and demonstrated the presence of an enhancer within its U3 sequence. There is evidence that trans-activation of the CAEV LTR requires U3 sequences. When the EIAV U3 region was replaced by the CAEV U3 sequence, the promoter activity of the EIAV LTR was markedly elevated, but the responsiveness to the EIAV trans activator could not be restored. PMID- 2552172 TI - Characterization of a simian hepatitis A virus (HAV): antigenic and genetic comparison with human HAV. AB - PA21, a strain of hepatitis A virus (HAV) recovered from a naturally infected captive owl monkey, is indistinguishable from human HAV in polyclonal radioimmunoassays and cross-neutralization studies. However, cDNA-RNA hybridization has suggested a significant difference at the genomic level between PA21 and a reference human virus, HM175. Further characterization of this unique HAV was undertaken in an effort to determine the extent of genetic divergence from human HAV and its relation to the conserved antigenic structure of the virus. The close similarity between PA21 and HM175 antigens was confirmed with an extended panel of 18 neutralizing murine monoclonal antibodies: a reproducible difference in binding to the two viruses was detected with only one antibody (B5 B3). The nucleotide sequence of the P1 region of the PA21 genome had only 83.2% identity with HM175 virus, a difference approximately twice as great as that found between any two human strains. Most nucleotide changes were in third base positions, and the amino acid sequences of the capsid proteins were largely conserved. Amino acid replacements were clustered in the carboxy terminus of VP1 and the amino-terminal regions of VP2 and VP1. These data indicate that PA21 virus represents a unique genotype of HAV and suggest the existence of an ecologically isolated niche for HAV among feral owl monkeys. PMID- 2552173 TI - Relative rates of RNA synthesis across the genome of Epstein-Barr virus are highest near oriP and oriLyt. AB - The rates of Epstein-Barr virus transcription were measured in isolated nuclei from marmoset and human lymphoblasts transformed in vitro. In B95-8, a marmoset B lymphoid cell line, the most frequently transcribed viral genes are the EBERs (small nuclear RNAs) and BHLF-1 (encoding a lytic-phase gene product). The EBERs and BHLF-1 genes are separated by nearly 50 kilobase pairs on the Epstein-Barr virus genome and lie adjacent to (less than 300 base pairs from) oriP and oriLyt, respectively. oriP and oriLyt are putative origins of viral DNA replication, and each is associated with a transcriptional enhancer element. Among the human B lymphoblastoid cell lines tested, only the transcription of EBERs predominates. PMID- 2552174 TI - Differential DNA binding of nuclear proteins to a long terminal repeat region of the MCF13 and Akv murine leukemia viruses. AB - Long terminal repeat (LTR) sequences of murine leukemia viruses (MLVs) have been demonstrated to be mainly responsible for the pathogenic differences in these retroviruses. A region of the LTR which is downstream of the enhancer elements has been shown to contribute both to enhancer activity as well as to disease specificity of MLVs. We have identified protein-DNA complexes generated by this region of a lymphomagenic MLV (MCF13) and one which is nonpathogenic (Akv). One protein-DNA complex we have observed for this region is unique to MCF13 DNA sequences. Detection of protein involved in this unique MCF13 complex in different cell lines revealed that it was ubiquitous. PMID- 2552175 TI - Nucleotide sequence of a neurovirulent variant of the type 2 oral poliovirus vaccine. AB - Infectious cDNAs of the Sabin type 2 poliovirus vaccine virus and a vaccine derived neurovirulent type 2 strain (P2/117) have been cloned in Escherichia coli. Nucleotide sequence analysis revealed that P2/117 differs from the vaccine strain by just 23 point mutations. Three occur in the 5' noncoding region. The remainder result in a total of 5 coding changes located in VP1, VP4, 2B, and 3D. The likely role of these mutations in the evolution to neurovirulence is discussed. PMID- 2552177 TI - Mutational analysis of bovine papillomavirus type 1 E5 peptide domains involved in induction of cellular DNA synthesis. AB - Early gene E5 of bovine papillomavirus type 1 encodes a 44-amino-acid protein whose expression can transform immortalized mouse cell lines. We have previously reported that a chemically synthesized E5 peptide functions to induce cellular DNA synthesis upon microinjection into growth-arrested mouse cells. We further defined the two E5 domains essential for the full DNA synthesis induction activity by the analysis of E5 deletion and amino acid substitution mutant peptides. The first domain is the C-terminal 13-amino-acid core which is sufficient to activate DNA synthesis at high peptide concentration and contains two essential, highly conserved cysteine residues. The second domain is the 7 amino-acid hydrophobic sequence contiguous to the core domain which is sufficient to confer a 1,000-fold higher molar specific activity to the E5 peptide. A random hydrophobic sequence, but not charged amino acids, fulfills the function of the second domain. PMID- 2552176 TI - A common site for immortalizing proviral integrations in Friend erythroleukemia: molecular cloning and characterization. AB - By using a tagged derivative of Friend spleen focus-forming virus, we previously obtained evidence that proviral integration(s) in the host genome can cause erythroblast immortality by abrogating the commitment of the cell to differentiate (C. Spiro, B. Gliniak, and D. Kabat, J. Virol. 62:4129-4135, 1988). Exploiting the fact that each leukemia was a single clone that contained one tagged provirus, we have now molecularly cloned and characterized one common genomic site for immortalizing proviral integrations. PMID- 2552178 TI - Duck hepatitis B virus can tolerate insertion, deletion, and partial frameshift mutation in the distal pre-S region. AB - In-frame and frameshift mutations were introduced into the pre-S region (1,212 base pairs) of duck hepatitis B virus. The in-frame mutants retained the inserted 12 nucleotides, while the frameshift mutants either reverted to wild type or exhibited a 10-nucleotide compensatory deletion downstream of the original mutation site. Thus, although duck hepatitis B virus has a compact and highly economical genome organization, it can replicate despite alterations of up to 9 amino acid codons in the pre-S and P open reading frames. PMID- 2552179 TI - Expression of the int-1 and int-2 loci in endogenous mouse mammary tumor virus induced mammary tumorigenesis in the C3Hf mouse. AB - The int-1 locus appears to be involved in over 80% of C3H exogenous mouse mammary tumor virus (MMTV)-induced mouse mammary tumors, and the int-2 locus appears to be involved in approximately 10% of these tumors. Analysis of 46 C3Hf mammary tumors resulting from endogenous, rather than exogenous, MMTV infection revealed that only 41% expressed int-1 RNA, while 2% expressed int-2 RNA. Our results suggest that in addition to the int-1 and int-2 loci, other loci may be involved in endogenous-MMTV-induced mammary tumors of the C3Hf mouse. PMID- 2552180 TI - Latent infections in spinal ganglia with thymidine kinase-deficient herpes simplex virus. AB - A herpes simplex virus type 1 variant [C239(TK-)] harboring a deletion in the thymidine kinase (TK) gene was assessed for capacity to establish latent infections. Outbred Swiss Webster mice were inoculated on both hind footpads, and numbers of neurons expressing latency-associated transcript and amounts of viral DNA in latently infected lumbosacral spinal ganglia were scored. C239(TK-) established levels of latent infection that were only slightly lower than those found for either a TK rescued variant of this agent or the parent wild-type KOS. However, in contrast to the TK+ viruses, C239(TK-) could not be reactivated when spinal ganglia were cultured in vitro. The results presented show that expression of the viral TK gene plays no major role in establishment of the latent state but that it functions during reactivation of latent virus from explanted ganglia maintained in vitro. PMID- 2552181 TI - cis-acting sequences that control the level of viral DNA synthesis in the polyomavirus late region. AB - A deletion in the polyomavirus late region results in a drastic reduction of viral replication, as shown after transfection of viral DNA into 3T6 cells. This mutation is cis acting, since cotransfection with wild-type DNA did not restore the normal phenotype. Viral DNA synthesis returned to normal levels only after reintroduction of the authentic sequences in either orientation. The data presented here suggest that these sequences are involved in the binding of a factor(s) that controls the level of viral replication. PMID- 2552183 TI - Wheat bran in the selective therapy of absorptive hypercalciuria: a study performed on 18 lithiasic patients. AB - The binding properties of raw vegetable fiber towards bivalent cations suggested the prescription of brain as a dietary supplement to limit intestinal calcium absorption in hypercalciuric patients. A group of 18 patients with a specific diagnosis of absorptive hypercalciuria received a dietary supplement of 14 gm. wheat bran at the 2 principal meals for 90 days. A complete assessment of mineral metabolism was performed after 45 and 90 days. Mean basal calciuria was 357 mg. per 24 hours and a significant decrease was noted after 45 days (245 mg. per 24 hours) and 90 days (240 mg. per 24 hours), with a p value of less than 0.01. Urinary oxalate did not vary significantly (0.34 to 0.38 to 0.31 mMol. per 24 hours) and neither did phosphate levels (1,020 to 900 to 893 mg. per 24 hours). A slight and pathologically insignificant decrease was noted in serum iron and urinary magnesium; this fact could be considered a side effect owing to the nonselective binding properties of fiber. Therefore, the positive results achieved confirm the effective action of wheat bran in the treatment of correctly diagnosed absorptive hypercalciuria. PMID- 2552182 TI - Effects of alterations in the leader sequence of Rous sarcoma virus RNA on initiation of translation. AB - The 372-nucleotide leader sequence of Rous sarcoma virus RNA contains three conserved short open reading frames and other sequences responsible for a variety of life cycle functions. We have investigated several aspects of the leader RNA which may influence the translation of the major coding regions to which the leader is juxtaposed. We found that small perturbations of the leader length do not affect the binding and scanning of ribosomal subunits by more than about 10%, that the length and/or structure of the RSV RNA leader is near optimal for translation of the major coding regions of the viral RNA, that inclusion or deletion of open reading frames influences downstream initiation in a manner that is not strictly additive, and that reinitiation of translation at the gag gene is very efficient. PMID- 2552184 TI - The relationship of deoxyribonucleic acid content to conventional prognostic factors in Wilms tumor. AB - Deoxyribonucleic acid ploidy as determined by flow cytometric analysis was compared to histological findings and tumor stage for accuracy of prediction of patient survival in a series of Wilms tumors. While anaplastic tumors were more likely aneuploid or tetraploid than diploid, no statistically significant relationship was found between deoxyribonucleic acid ploidy and patient survival in our series. Tumor histology and stage as defined by the National Wilms Tumor Study were more accurate predictors of patient survival than deoxyribonucleic acid ploidy. PMID- 2552185 TI - Innovative ways to fight cancer dominate 1989 AMA Houston science news conference. PMID- 2552186 TI - Anti-HIV substances for rape victims. PMID- 2552187 TI - From the National Institutes of Health. PMID- 2552188 TI - [Analyses of pathogenic factors in bacteria--bacterial toxins: clostridial toxins, e.g. C. tetani neurotoxins and C. perfringens type A enterotoxin]. PMID- 2552189 TI - Calcium-activated neutral protease inhibitor (E-64c) and reperfusion for experimental myocardial infarction. AB - We examined the efficacy of the combination of coronary reperfusion and calcium activated neutral protease (CANP) inhibitor (E-64c) for the treatment of acute myocardial infarction in dogs. In 34 dogs, the left anterior descending artery (LAD) was occluded and reperfused after 1 hour (Groups A and B). In the remaining 49 dogs, the LAD was ligated (Groups C and D). E-64c (100 mg/kg, Groups A and C) or vehicle (Groups B and D) was injected intravenously before and after the coronary occlusion or ligation. After 24 hours the hearts were removed. The proportion of the infarct size in the LAD perfusing area (risk zone) in Group A was 47.3 +/- 9.7%, significantly lower than in Group C (54.8 +/- 8.2%, p less than 0.05) or Group D (58.7 +/- 10.0%, p less than 0.01). There was a significant difference between Group B (52.9 +/- 8.6%) and Group D as well (p less than 0.05). The effects of reperfusion (p = 0.0016) and E-64c (p = 0.0226) per se on infarct size were significant, but the combination of reperfusion and E-64c was not additive. The decrease in CPK activity in the risk zone was significantly lower in the reperfused group (p = 0.0001). The mCANP activity was higher in the border zone and lower in the infarct zone. The trend in the mu CANP activity was similar to that of mCANP. Thus, treatment with a CANP inhibitor in the early phase of acute myocardial infarction may be marginally beneficial in combination with reperfusion. PMID- 2552190 TI - Positive inotropic and negative chronotropic effects of OPC-8490, a newly developed cardiotonic, in isolated, blood-perfused canine heart preparations. AB - When a new cardiotonic, OPC-8490 (3, 4-dihydro-6-[4-(4-oxo-4-phenylbutyl)-1 piperazinylcarbonyl]- 2(1H)- quinolinone citrate), was administered into the jugular vein of a support dog, at doses of 0.1, 0.3 and 1 mumol/kg, decreases in heart rate and arterial blood pressure were dose-dependently induced in intact support dogs. One and a half min after administration, positive inotropic and slight negative chronotropic responses were observed in isolated right atria perfused with arterial blood of support dogs. Administration of OPC-8490 into the sinus node artery of the isolated atrium induced positive inotropic and biphasic chronotropic effects, an initial brief positive (Ph1) followed by a long-lasting negative (Ph2) chronotropic effect in a dose-related manner. OPC-8490 at 1000 nmol caused a triphasic, Ph1 followed by Ph2 and slight positive (Ph3) chronotropic effects and an inotropic effect. In the left ventricular muscle preparation driven electrically at 2 Hz, 10-3000 nmol of OPC-8490 increased contractile force in a dose related manner. OPC-8490-induced responses were not significantly modified by propranolol or atropine. When isoproterenol (0.04 nmol/min) increased the basal sinus rate and contractile force of isolated atria, the Ph1 was suppressed and the Ph3 became clear, although the negative phase (Ph2) was not changed. The positive inotropic effect was not significantly changed. When intramural vagal nerve stimulation decreased sinus rate and contractile force, the positive inotropic and negative (Ph2) chronotropic effects were depressed. Verapamil significantly depressed the positive inotropic but not the chronotropic responses to OPC-8490. The positive inotropic effect of OPC-8490 was depressed by pinacidil but not changed by ouabain, although the chronotropic responses to OPC-8490 were not changed. These results suggest that cardiac responses to OPC-8490 involve several mechanisms including cyclic AMP dependent, Ca channel-dependent, potassium current-inhibitory and/or other mechanisms in the dog heart. PMID- 2552191 TI - Distribution and metabolism of norepinephrine, cyclic AMP and cyclic GMP in the atrioventricular conducting tissue of the bovine heart. AB - The distribution of norepinephrine (NE), cyclic AMP (cAMP) and cyclic GMP (cGMP) and the activities of related enzymes in the atrioventricular (A-V) conducting tissue of the bovine heart were examined. The concentration of NE in the atrium was about twice that in the ventricle. In the A-V conducting tissue, the concentration of NE was highest in the atrioventricular node (AVN) and lowest in the false tendon (FT), with intermediate levels in the bundle of His (HIS) and the right and left bundle branches (RLBB). The activity of monoamine oxidase (MAO) in the atrium was about 2.2 times that in the ventricle. In the A-V conducting tissue, the activity of MAO was highest in the HIS and lowest in the FT. The activity of catechol-o-methyltransferase (COMT) in the atrium and ventricle was similar, and that in the HIS was slightly, but not significantly, higher than that in other regions of the A-V conducting tissue. The concentration of cAMP in the ventricle was about twice that in the atrium. In the A-V conducting tissue, the concentration of cAMP was higher in the AVN and FT than in the HIS and RLBB. The distribution of adenylate cyclase (AC) was similar to that of NE. The phosphodiesterase (PDE) activity in the atrium and ventricle was similar. No significant difference was found in the level of PDE activity in different regions of the A-V conducting tissue. The concentration of cGMP was slightly, but not significantly, higher in the A-V conducting tissue than in the atrium or ventricle. In the A-V conducting tissue, the concentration of cGMP was highest in the FT and the concentrations in the HIS, RLBB and AVN were similar. These findings suggest that in the A-V conduction tissue, the regions that have the higher spontaneous pacemaker rates have higher NE content and AC activity, that is sensitivity to NE. Furthermore, the sensitivity for muscarinic cholinergic stimulation is higher in the conducting tissue (especially in the FT) than in the atrium and ventricle. PMID- 2552192 TI - [Hodgkin's disease diagnosed from the numerous Reed-Sternberg cells in the bone marrow]. AB - A 46-year-old female was admitted to our hospital for fever and weight loss in September, 1986. Physical examinations were unremarkable. CBC revealed moderate anemia and leukopenia with abnormal lymphocytes. Examination of the bone marrow (BM) disclosed peroxidase negative blasts and multinucleated or multilobulated giant cells positive for CD30 (Ki-1) antigen. Chest X-ray was negative. CT scan and echography of the abdomen showed minimal enlargement of retroperitoneal lymph nodes (LN). Lymphangiography revealed mild enlargement of the LN without filling defects. Gallium scan was negative. Hence a diagnosis of Hodgkin's disease (HD) stage IVB was made. She was treated with MOPP therapy with modification and obtained a complete remission. BM involvement of HD occurs mostly in advanced stages. We assumed that this is a rare case of HD in which bone marrow metastasis occurred in a very early stage of the disease or that of bone marrow primary. PMID- 2552193 TI - [Impaired superoxide production by neutrophils in myelodysplastic syndrome: correlation with bone marrow karyotype]. AB - Superoxide (O2-) production by neutrophils was examined in 21 patients with myelodysplastic syndrome (MDS), including 13 with primary MDS (p-MDS) and 8 with therapy-related MDS (t-MDS). O2-production of MDS patients was significantly less than that of healthy controls (2.81 +/- 3.51 vs 6.19 +/- 2.41 nmol/min/10(6) PNMs, p less than 0.005). Although no relationship between pathogenesis (primary or therapy related), FAB type and levels of O2- production was observed, O2- production of 11 patients with abnormal bone marrow karyotypes was significantly reduced than that of 10 other patients with normal karyotypes. Furthermore, five t-MDS patients with monosomy for all or part of chromosome no. 7 (-7 or 7q-) showed the lowest level of O2- production, which was significantly different from the level in the patients with other abnormal karyotypes (0.61 +/- 0.29 vs 1.80 +/- 0.31, p less than 0.001). These results suggest that the long arm of chromosome no. 7 may in part play an important role for O2- production by neutrophils. PMID- 2552194 TI - [Neurofibroma-associated left peroneal nerve palsy in a patient with acute lymphoblastic leukemia]. AB - Peroneal nerve palsy developed in a patient with T cell-type acute lymphoblastic leukemia (ALL) is reported. In the fifth month after starting of chemotherapy against ALL, the patient, a 7-year-old girl, developed drop foot on the left. Three possibilities were considered as its pathogenesis; (1) VCR neuropathy, (2) neurotoxicity of intrathecal MTX, (3) leukemic invasion to the spinal canal. However, there was no evidence of leukemic invasion in any lumbar taps, and no improvement was obtained by cessation of VCR and intrathecal MTX. Examination by CT scan revealed tumors in the intervertebral (L5-S2) region, which was diagnosed to be neurofibromas by biopsy. The tumors compressed the left peroneal nerve and neurotoxicity of antineoplastic agents for ALL could be the cause of her drop foot. PMID- 2552195 TI - [DS-bronchial arteriography; demonstration of metastasized lymphnodes of lung cancer--with special reference to the primary effect in BAI]. AB - We analysed the 44 DS-Bronchial arteriograms (DSBAG) of lung cancer with an attention to the demonstration of metastasized mediastinal lymphnodes. The stain of lymphnode is well demonstrated in the pericarinal region, such as station #7, #R4 along the course of bronchial arteries. The swollen nodes with stains on DSBAG show the good reduction rates (37.5%) after BAI that is analogous to the primary lesion (38.9%), in contrast to the nodes without stains (24.5%). DSBAG with excellent contrast resolution contributes to the BAI as verifying the infused area. PMID- 2552196 TI - [A histological examination of liver fibrosis and tissue prolyl hydroxylase in various liver diseases]. AB - Prolyl hydroxylase (PH) is an enzyme acting in early stage of collagen synthesis. We have emphasized the significance of the measurement of serum PH (SIRPH) in relation to liver fibrosis mainly in patients with alcoholic liver disease (ALD). In this study, we determined the localization and positivity of PH by tissue PH stain method (Avidin Biotin Complex method) to clarify the differences in fibrosis between ALD (25 cases) and non-alcoholic liver diseases (non-ALD, 47 cases). Tissue PH was found to be positive in liver cells around Glisson's sheath in early stage of fibrosis, and then in liver cells left within septa and also in mesenchymal cells in the sinusoidal wall as fibrosis progressed. Although there were basically no marked differences between ALD and non-ALD, ALD tended to show stronger tissue PH positivity for a degree of fibrosis, PH positivity in parenchymal cells was especially remarkable around pericellular fibrosis in ALD. These results clearly reflected the important role of liver parenchymal cells in progression of fibrosis. PMID- 2552197 TI - [Imaging diagnosis of ruptured site in hepatocellular carcinoma]. AB - Rupture causes frequently the patients with HCC to death. Then it is important to investigate ruptured site of HCC in order to stop bleeding effectively and safely by emergent TAE without causing liver failure. We evaluate the usefulness of 3 major imaging method, such as US, CT and angiography in our 18 cases of HCC. In ultrasonography, ruptured site was shown as the high echoic area localized around the tumor in 4/15 cases (26.7%). In computed tomography of rapid scanning, it was shown as the intraperitoneal high density area around the tumor in 3/18 cases (16.7%). And lipiodol CT showed the characteristic leakage in the abdominal cavity. In angiography, it was shown as the extravasation of contrast material only in 2/13 cases (15.4%). In the results, it was possible to diagnose the ruptured site in 8/18 cases (44.4%) by using these procedures. Diagnosis of ruptured site will make it possible to reduce the frequency of hepatic failure caused by emergent TAE. PMID- 2552198 TI - [A case of early gastric carcinoma with disseminated carcinomatosis of bone marrow]. PMID- 2552199 TI - [New color imaging of [99mTc]-pyrophosphate and [201Tl]-chloride dual isotope single photon emission computed tomography in acute myocarditis]. AB - [99mTc]-pyrophosphate (PYP) and [201Tl]-chloride dual isotope single photon emission computed tomography (SPECT) is now available to detect the site and extent of acute myocardial infarction. In inflammatory myocardial disease, [99mTc]PYP makes hot image on damaged area. We performed dual isotope SPECT of [99mTc]PYP and [201Tl]Cl in two patients with acute myocarditis and severe rhythm disturbance to evaluate the severity of inflammation. Myocardial damage was estimated by [201Tl] perfusion coloring blue and myocardial inflammation was estimated by [99mTc]PYP uptake coloring red. The overlap display of both images made it clear to detect spatial extent of myocardial inflammation. Using this technique, we expect to estimate the severity of myocarditis and to make a decision of therapeutic plan. PMID- 2552200 TI - Influence of sulfhydryl agents on cytoskeleton in cultured human trabecular cells. AB - The effects of three sulfhydryl agents on the cytoskeletal systems of cultured human trabecular cells were examined by the immunofluorescence method. Incubation with iodoacetamide or N-ethyl maleimide (10(-6) M, 10(-5) M, and 10(-4) M) caused a marked alteration on the actin and microtubule organization, but had little effect on the structural integrity of the vimentin intermediate filaments. Incubation with p-chloromercuribenzene sulfonate (10(-6) M, 10(-5) M, and 10(-4) M) caused no remarkable changes in the three major cytoskeletal systems. These findings suggest that the trabecular cell cytoskeleton plays an important role in increasing the aqueous outflow facility after treatment with the sulfhydryl agents, iodoacetamide and N-ethyl maleimide. PMID- 2552202 TI - An investigation on the inactivation process of sodium currents in single frog ventricular cells. AB - The characteristics of sodium currents (INa) in single frog ventricular cells were studied with the oil gap method. This method improves time- and space control of the membrane potential under the voltage clamp, thereby making possible accurate analysis of fast events of INa. In this preparation the threshold of INa was about -60 mV and the reversal potential was 58 mV, which is close to the value calculated by the Nernst equation for sodium ions. Because the instantaneous current-voltage (I-V) relationship is linear, the ease of permeation of sodium ions through Na+ channels is well expressed by the chord conductance. The falling phase of INa and the time course of recovery from inactivation follow a time course of single exponential function. The time constants for on- and off-processes of inactivation at the same membrane potential are very close to each other, indicating only a single state of inactivation. Though almost all properties of INa were well described by Hodgkin Huxley's model, a clear delay of onset of inactivation was demonstrated by two pulse experiment. In this report the modified kinetics scheme was proposed which can account for both a delay of onset of inactivation development and a falling phase of INa that follows a single exponential time course. PMID- 2552201 TI - Characterization of peptide inducing cataractogenesis in lens of hereditary cataractous rat (ICR/f RAT). AB - A material with inhibitory action to Na+/K+ ATPase was found in the lens of the ICR/f rat, a recessive hereditary cataractous rat. The material also induced lens opacification in vitro. From the results of amino acid analysis and by secondary ion mass spectroscopy, it was suggested that the material might contain approximately equimolar amounts of four amino acids, ie, aspartic acid, serine, glutamic acid and glycine, and that the molecular weight was 444. These facts suggested that this material with Na+/K+ ATPase inhibitory action might be a peptide. However, there is not yet any corroborating evidence to show whether this peptide is only a single material or not. The peptide significantly increased with aging in the lens of the ICR/f rat until approximately 90 days, when cataract became manifest, but its content decreased thereafter. This study suggests that one of the causes of cataractogenesis in the ICR/f rat might be this peptide, which is transformed in the lens with aging, and also that the peptide might accelerate lens opacification after cataractogenesis. PMID- 2552203 TI - Dual regulatory mechanisms of proton transport in rat papillary collecting duct cells in culture. AB - The regulation of proton transport and cytosolic pH was studied in rat papillary collecting duct (PCD) cells in culture using a pH-sensitive fluorescence probe, 2,7-bis-carboxyethyl-5,6-carboxyfluorescein (BCECF). Data were obtained from confluent monolayers grown on glass coverslips and dipped in a HCO3- -free medium, pH 7.40. The resting intracellular pH (pHi) was 7.16 +/- 0.03 (n = 20). When PCD cells had been acidified by pretreatment with NH4Cl, pHi immediately recovered toward the resting value. Two mechanisms participated in this recovery: a Na+-dependent mechanism which could be inhibited by amiloride (indicative of Na+-H+ exchanger) and a Na+-independent process (a proton ATPase). The pHi recovery from acid loading was inhibited by amiloride to about 55% of the control recovery (half-maximal effect at 100 microM). The rate of pHi recovery after the readdition of Na+ to a sodium-free medium exhibited saturation kinetics (half maximal rate at 28 mM). Dicyclohexylcarbodiimide (DCCD), an inhibitor of a plasma membrane proton ATPase, and the depletion of cellular ATP induced by 2 mM potassium cyanide (KCN) also partially inhibited the rate of pHi recovery after cell acidification with a NH4Cl load. When PCD cells were treated with 1 mM DCCD, amiloride almost completely inhibited pHi recovery. Amiloride and the removal of external Na+ had induced a gradual fall in pHi to a new resting value and rapidly recovered when Na+ was added. We conclude that PCD cells grown in culture have at least two proton transport mechanisms: a Na+-H+ exchanger and a plasma membrane proton ATPase. The kinetics of these processes can be reliably assessed by the pH sensitive fluorescent probe, BCECF. Both the Na+-H+ exchanger and the plasma membrane proton ATPase may contribute to urinary acidification. PMID- 2552204 TI - Time dependence of Li+ action on acetylcholinesterase activity in correlation with spontaneous quantal release of acetylcholine in rat diaphragm. AB - The mean miniature end-plate potential (m.e.p.p.) frequency and the acetylcholinesterase (AChE) activity have been examined in the rat diaphragm, in relation to the time of Li+ action. Substitution of LiCl for NaCl caused increase in m.e.p.p. frequency, the final level of which depended on Li+ concentration in the bath solution. Incubation of diaphragm homogenate with NaCl caused initially a slight increase of AChE activity, reaching a plateau after 20 min. Substitution of LiCl for NaCl produced an increase in AChE activity to a maximum, followed by a decrease to a minimum. The activity consequently returned to a steady level. During complete substitution of LiCl for NaCl, the maximum value of AChE activity coincided with the maximum m.e.p.p. frequency (Fmax) induced by Li+; the minimum value correlated with the precise instant that the frequency reached a plateau. The addition of prostigmine caused: a) blockade of AChE activity, and b) prostigmine concentration-dependent decrease of Fmax and/or disappearance of m.e.p.p.s from neuromuscular junctions treated with Li+. These results are discussed in terms of correlation between changes of AChE activity induced by Li+ and variations of spontaneous quantal release of the transmitter from nerve terminals of the rat diaphragm treated with Li+. PMID- 2552205 TI - Beta-adrenergic receptor adaptation after an acute exercise in rat myocardium. AB - An acute dynamic exercise provokes the translocation of beta-adrenergic receptors (beta-AR) from light vesicle fractions to sarcolemmal membranes in rat myocardium. However, 15 min after an acute exercise the density of beta-AR in both fractions returned to the pre-exercise level. The mean maximal activity of adenylate cyclase in response to isoproterenol roughly paralleled the redistribution of beta-AR. The dose-response curves, however, were substantially shifted to the right with increase in EC50 for isoproterenol stimulation of adenylate cyclase. Thus, the sensitivity of sarcolemmal beta-AR was found to be blunted 15 min afterwards. PMID- 2552206 TI - Depression of voltage-dependent Ca2+ current by noradrenaline in the guinea-pig vas deferens in tetraethylammonium medium. AB - The actions of noradrenaline (NA) on the smooth muscle of the guinea-pig vas deferens in tetraethylammonium (TEA) medium have been studied under constant current and voltage-clamp conditions. The marked changes observed during exposure to NA are 1) slowness in the rising phase of the action potential, delay in the falling phase and reduction in the amplitude, and 2) depression of the voltage dependent Ca2+ current with a negative shift of its reversal potential as well as with a reduction in its conductance. These findings might suggest that NA application stimulates the voltage-independent Ca2+ machinery, but suppresses the voltage-dependent Ca2+ machinery through membrane. PMID- 2552207 TI - Histopathological study on a case of idiocy--morphological mechanism of idiocy appearance. AB - A male, aged 42, had idiocy without any somatic or neurological signs. In laboratory examination, no abnormal finding could be observed. Brain weight was 1,330 g. and no abnormal finding was observed macroscopically. Histopathologically, gross changes were not observed, however, many nerve cells contained a lot of lipofuscin granules in the cytoplasms for the patient's age. Some astrocytes in the cerebral cortex contained homogenous amorphous inclusion bodies in their cytoplasms. From these findings, it was speculated that the change of astrocyte may play an important role in causing mental deficiency. PMID- 2552208 TI - Further postmortem examination of a case of familial ataxia with cerebrospinal fluid abnormality: an electron microscopic study of the intracytoplasmic eosinophilic inclusion bodies in the central nervous system. AB - As reported previously, the peculiar intracytoplasmic eosinophilic inclusion bodies (IEIBs) extensively appeared in the autopsied brain tissue from a 49-year old man having familial ataxia with cerebrospinal fluid abnormality, and histochemically showed abundant proteins, but few lipids and carbohydrates. Ultrastructurally, many membrane-bound vacuoles derived from the distended cisterns of rough-surfaced endoplasmic reticulum (RER) appeared in the neurons. They were filled with fine granular, less dense materials. The IEIBs, shown as a homogeneous dense core, were found in some of the vacuoles. Similar vacuoles also appeared in astrocytes, oligodendrocytes, vascular pericytes, ependymal and choroidal epithelial cells. It is suggested that the vacuoles result from the accumulation of metabolic products in the distended RER cisterns of the cells in the central nervous system, presumably representing a genetically determined functional abnormality of the RER in protein synthesis and/or transport. PMID- 2552210 TI - [A case report of small cell lung cancer with extrahepatic biliary obstruction and intramedullary spinal cord metastasis]. AB - A 62-year-old man was admitted to our hospital because of high serum amylase and neck swelling. His chest radiography revealed a mass lesion in the right upper lung. Open lung biopsy specimen established the pathological diagnosis as intermediate type small cell lung cancer. In his clinical course, obstructive jaundice recurred several times with response to combination chemotherapy. The findings of abdominal echogram, computed tomography and endoscopic retrograde cholangiopancreatography suggested that the jaundice was caused by extrahepatic biliary obstruction with parapancreatic lymph node metastasis. He died of spinal bleeding which spread from the medulla oblongata to the thoracic spinal cord. Bleeding was caused by intramedullary spinal cord metastasis. Both extrahepatic biliary obstruction and intramedullary spinal cord metastasis are rare and their symptoms were more prominent than those of the pulmonary lesion. PMID- 2552209 TI - [Measurement of carcinoembryonic antigen, squamous cell carcinoma related antigen and neuron-specific enolase in the bronchoalveolar lavage fluid in patients with peripheral lung cancer]. AB - Carcinoembryonic antigen (CEA), squamous cell carcinoma related antigen (SCC) and neuron-specific enolase (NSE) in bronchoalveolar lavage fluid were measured in 30 patients with peripheral lung cancer, 11 patients with benign lung disease and 19 healthy controls. The mean levels and positive rates of lavaged fluid CEA were 128.0 +/- 16.9 ng/mg and 33.3% in patients with lung cancer, 68.1 +/- 25.9 ng/mg and 9.1% in patients with benign lung disease, and 68.3 +/- 11.6 ng/mg and 5.2% in healthy controls, respectively. The mean levels and positive rates of lavaged fluid CEA in patients with lung cancer were significantly higher than those in patients with benign lung disease (p less than 0.05) and those in healthy controls (p less than 0.05). The mean levels of lavaged fluid SCC and NSE showed no significant difference between cases of lung cancer and benign lung disease or healthy controls. No lavaged tumor marker level in patients with lung cancer showed any close correlation with histologic types and serum levels. In conclusion, measurement of lavaged fluid CEA was considered to be useful in the differential diagnosis of peripheral lung cancer. PMID- 2552211 TI - [Carbohydrate antigens in pleural effusion from patients with lung cancer]. AB - Sialosylated Lewisa (S-Lea) is an active antigen determinant of CA19-9 which is well known as a tumor maker. Sialosylated Lewisx (S-Lex) is a positional isomer of S-Lea. The positive percent of S-Lex and CA19-9 in pleural effusion from the patients with primary lung cancer was 49.1% and 40.0%, respectively. However, most S-Lea positive effusions showed a positive level of S-Lex. Therefore, S-Lex is considered to be more useful as a tumor marker than CA19-9. The studies on gel chromatography of the effusion obtained from a patients showed high levels of both antigens using sephacryl S-1000 revealed that the molecular weights of S-Lex and CA19-9 were more than 2 X 10(6) and the elution profile of S-Lex coincided with that of CA19-9 fractionated effusions using the monoclonal antibodies to the antigens showed the same pattern of elution profiles of these carbohydrates. From these results, we concluded that the carriers of S-Lex and CA19-9 in pleural effusion from lung cancer might be the same molecules with a high molecular weight more than 2 X 10(6). PMID- 2552212 TI - [An immunoradiometric assay of the serum TPA from patients with primary lung cancer and a comparison with the conventional double antibody method]. AB - The serum level of tissue polypeptide antigen (TPA) has been measured by a new immunoradiometric assay (IRMA) method that has been recently developed using a specific antibody, and compared with the results obtained by the conventional double antibody method, using a polyclonal antibody. Both methods gave essentially the same results, i.e. the serum TPA level in patients with primary lung cancer was higher when compared with non-malignant pulmonary diseases and healthy individuals (p less than 0.01). However the positive rate and average values of TPA with patients of non-malignant diseases were found to be lower by the new IRMA method than by the conventional method. This may be due to the improvement in specificity and accuracy of this newer method of assay, as they also statistically decrease within the cut off level (110 U/L) estimated in non malignant diseases and healthy individuals. Therefore, it has been concluded that the IRMA method is more suitable than the conventional method for diagnosis of primary lung cancer. PMID- 2552213 TI - [An autopsy case of pulmonary asbestosis with synchronous multiprimary lung cancers, early gastric cancer and atypical mycobacteriosis]. AB - An autopsy case of pulmonary asbestosis with synchronous multiprimary lung cancers, gastric cancer and atypical mycobacteriosis of a 64-year-old asbestos textile worker was reported. Chest X-ray film on the first admission showed a coin lesion on the left upper lung field, and on the second admission showed a progressively enlarging shadows in the left hilum. Chemotherapy showed almost no effect and the patient died of respiratory failure. Autopsy revealed moderate asbestosis, with triple lung cancers (squamous cell carcinoma, adenocarcinoma and small cell carcinoma) in the left upper lobe, gastric cancer (signet-ring cell carcinoma) and atypical mycobacteriosis. So far as we know, no case with such complex lesions has ever been reported. PMID- 2552214 TI - [Mediastinal dissection of hepatocellular carcinoma with bilateral hilar and mediastinal lymph node metastasis]. AB - A 54 year-old man, who had been operated for hepatocellular carcinoma 43 months before, complained of frequent cough. He had widening of mediastinal shadow on chest X-ray film and high level of alpha-fetoprotein (197 ng/ml). Under the diagnosis of metastasis to mediastinum of hepatocellular carcinoma, lymph node dissection was performed and disclosed large metastatic tumors in subcarinal, right tracheobronchial and left hilar lymph nodes. After operation, frequent cough disappeared and alpha-fetoprotein level fell to 38 ng/ml. Metastasis to the mediastinum without lung metastasis of hepatocellular carcinoma was recognized only 2.6% in non-operated cases in annual of pathological autopsy cases in Japan 1980 and 4 case reports were found in literature with large metastatic mass shadow in mediastinum of hepatocellular carcinoma. PMID- 2552215 TI - [Influence of dietary animal protein on renal stone disease]. AB - The effect of dietary protein load on the incidence of nephrolithiasis was studied in rats and men. Three groups of adult male Wister rats were fed with a standard protein diet, a high protein diet, or a low protein diet for 4 weeks. In the high protein group, calcium excretion was significantly increased and citrate excretion was remarkably decreased. This group also exhibited low grade metabolic acidosis due to catabolism of excess amino acids, and increases in urinary cyclic AMP excretion and bone resorption. These findings indicate that protein-induced hypercalciuria is due to low grade metabolic acidosis, which directly affects renal handling of calcium. Long-term calcium loss in the urine may lead to negative calcium balance and hyperfunction of the parathyroid gland may induce bone resorption. The influence of 40 g animal protein load on urinary risk factors of calcium stone formation was investigated in 23 healthy males and 26 patients with nephrolithiasis. All subjects were given control diets each day containing 60 g protein for a week and during the next week each received an additional 40 g animal protein. In the controls, added dietary protein resulted in decreased urinary citrate and increased urinary uric acid, with no change in urinary calcium or cyclic AMP excretion. In contrast, the patients showed increased urinary calcium and cyclic AMP as well as decreased urinary citrate. Further examination of the patients revealed that the significant increases of calcium and cyclic AMP excretion occurred only in hypercalciuric patients, who seemed to be classified into renal hypercalciuria.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552216 TI - [Evaluation of renal scarring in children with primary VUR by 99mTc-DMSA renoscintigraphy]. AB - Renal scarring in 271 kidneys of 172 children with primary vesicoureteral reflux (VUR) was evaluated by 99mTc-DMSA renoscintigraphy. 58% of refluxing kidneys were with renal scar by the initial DMSA renoscintigraphy. Only 52% of these kidneys showed good correlation between the findings on IVP and DMSA renoscintigram. Of the 144 refluxing kidneys with normal IVP, 41% had renal scarring on DMSA renoscintigram. DMSA renoscintigram revealed widespread renal scarring in 28% of kidneys with only calyceal clubbing and in 60% of those with segmental cortical thinning on IVP. It is realized that IVP was an in-sensitive method to evaluate renal scarring of refluxing kidneys and such kidneys with segmental renal scar on IVP accompanies more widespread scar on DMSA renoscintigram. These cases were allocated to 2 age groups, younger than 3 years and older than 4 years. In the former group less than 10% of kidneys with low grade VUR and about 40% with high grade UVR had widespread renal scarring. On the contrary, in the latter group severe renal scar was recognized in more than 20% of kidneys with low grade VUR and in about 60% with high grade UVR. PMID- 2552217 TI - A systematic program designed to provide trauma patient follow-up to emergency nurses. PMID- 2552219 TI - [Effects of hemosorption on the level of myoglobin, ACTH and cortisol in the blood of patients with ischemia of the lower limbs]. PMID- 2552218 TI - [Role of biochemical and radiological study methods in the evaluation of prognosis in acute ischemia of the limbs]. AB - Examined were 47 patients with acute ischemia of the extremities: 29--with favourable outcome, and 18-with the development of gangrene. Scintigraphy of the extremity with 99mTc-Sn-pyrophosphate was performed; urinary excretion of epinephrine and norepinephrine, DOPA and dopamine, and vanillylmandelic acid as well was studied; the content of ACTH, cortisol and cyclic adenosine monophosphate in the blood was defined. The use of radionuclide and biochemical methods of investigation permits to assess the degree of severity of the ischemic injury to the tissues of the extremities. PMID- 2552221 TI - [Phrenic nerve paralysis after heart operations]. PMID- 2552220 TI - [One-step breast reconstruction]. AB - The mental trauma induced by removal of the mammary gland for cancer may be overcome by reconstruction of the breast. One-step reconstruction applied on 4 patients showed considerable advantages over a postponed reconstruction: technically it is easier to perform; a single stay of the patient in hospital makes it cheaper. It is no obstacle to perform pre- and postoperative radio- and chemotherapy. According to available data in the literature, implanted prosthesis or transposed flap do not veil a local relapse. PMID- 2552223 TI - [Experiences with a "rapid test" in detecting antibodies to cytomegalovirus]. AB - Transmission of cytomegalovirus (CMV) to CMV negative transplant or blood recipients should be avoided in organ transplantations, since CMV infection of immunocompromised patients can result in severe illness. The infectivity of organ and blood donors is judged on the basis of CMV antibody, considering CMV IgG positives as potentially infectious. Several sensitive test systems are available for testing organ and blood donors for CMV IgG antibody which should be carried out prior to each transplantation. A series of sera were tested comparatively for CMV IgG antibody by latex agglutination test, ELISA and immunofluorescence. It was found that the rapid and easily performed latex agglutination test was much less sensitive than were the other techniques employed. As a consequence, positive latex agglutination test results are acceptable, only, while negative results should be controlled by more sensitive methods. PMID- 2552222 TI - The adrenergic system in lymphocytes from children with cystic fibrosis. AB - Several in vivo and in vitro studies have suggested that children suffering from cystic fibrosis (CF) might have a general defect of beta-adrenoceptors on the cell surface which might account for an unbalanced secretory process. In order to investigate if this view holds true, we determined the beta-adrenoceptor density and affinity on lymphocytes by means of radioligand studies using 125-iodo-cyano pindolol (125-ICYP) in 20 children with CF. Cyclic AMP (cAMP) response was also investigated after specific beta-adrenoceptor stimulation with isoprenaline (IPN) and after direct stimulation of the adenylate cyclase with forskolin in lymphocytes. Children with CF and controls have identical numbers and affinities of beta-adrenoceptors on lymphocytes. The cyclic AMP response was identical in CF and in age-matched control children regardless whether adenylate cyclase was stimulated directly or via beta-adrenoceptors. In conclusion, the data support the view that no general adrenoceptor or adenylate cyclase defect exists in CF. As several studies have found abnormal reactions to adrenergic stimuli in CF patients, we presume that there is a defect beyond the level of adrenergic receptors and cAMP which remains to be identified. PMID- 2552224 TI - Distribution of merosin, a laminin-related tissue-specific basement membrane protein, in human Schwann cell neoplasms. AB - The expression of merosin and laminin was studied in human schwannomas, plexiform neurofibromas, and malignant schwannomas immunohistochemically using monoclonal antibodies. Merosin is a unique novel tissue-specific basement membrane protein found in basement membranes of trophoblast, striated muscle, and Schwann cells. Merosin is related to laminin, another basement membrane protein with which it shows a homologous C terminal domain. In schwannomas, merosin was only found in areas where tumor cells were in contact with stromal or vascular tissue. Laminin, however, was present in all tumor cell basement membranes. In plexiform neurofibromas large amounts of both merosin and laminin were seen in Schwann cell basement membranes. Very little of either protein was found in malignant schwannomas. Thus merosin is present almost exclusively in highly differentiated Schwann cell neoplasms, and its distribution is more restricted than that of laminin. The expression of merosin in plexiform neurofibromas and in the schwannoma cells juxtaposed to the mesenchymal cells suggests that this protein is induced epigenetically in well-differentiated cells in contact with connective tissue or vascular components. PMID- 2552225 TI - Comparison of arachidonic acid metabolism by pulmonary intravascular and alveolar macrophages exposed to particulate and soluble stimuli. AB - Pulmonary intravascular macrophages, as prominent components of the pulmonary mononuclear phagocyte system, could be significant mediators of lung inflammation. We have shown that intravascular and alveolar macrophages metabolize exogenous arachidonic acid to its inflammatory metabolites via the lipoxygenase and cyclooxygenase pathways after exposure to the calcium ionophore A23187. In this study, we compare the metabolism of endogenous arachidonic acid by porcine intravascular and alveolar macrophages after exposure to soluble and particulate stimuli. Since intravascular and alveolar macrophages are exposed to various stimuli in vivo, it is essential to know the range of inflammatory mediators that these cells can generate. Alveolar macrophages attached to plastic and exposed to the various stimuli produced prostaglandin F2 alpha, 12 hydroxyheptade-catrienoic acid (HHT), hydroxyeicosatetraenoic acids (HETE), and leukotriene B4. In contrast, adherent and stimulated intravascular macrophages produced several cyclooxygenase products and lipoxygenase products including 5 HETE, 12-HETE, and leukotriene B4. Both macrophages released large amounts of arachidonic acid upon exposure to each stimulant. Intravascular macrophages that were adherent to plastic or were stimulated with glass, asbestos, or A23187 released significantly (p less than 0.05) more metabolized arachidonic acid than similarly treated alveolar macrophages. The major cyclooxygenase metabolite released by alveolar macrophages was prostaglandin 2 alpha, whereas HHT was the primary metabolite of intravascular macrophages. The major lipoxygenase metabolite released by both macrophage types was 5-HETE, but intravascular macrophages also released substantial amounts of 12-HETE and leukotriene B4. In both macrophage preparations, lipoxygenase products composed most released metabolites. After exposure to iron, asbestos, and A23187 intravascular macrophages released significantly more (p less than 0.05) lipoxygenase metabolites than alveolar macrophages. However, in alveolar macrophages, chrysotile asbestos induced greater activity by the cyclooxygenase pathway than by the lipoxygenase pathway. Both asbestos and iron spheres induced release of arachidonic acid and its metabolites, but the most potent stimulants in both macrophage preparations were A23187, zymosan, and lipopolysaccharide. We conclude that stimulated intravascular macrophages use both cyclooxygenase and lipoxygenase pathways to metabolize endogenous arachidonic acid, that these macrophages are metabolically more active than alveolar macrophages, and that both macrophage types are induced to metabolize arachidonic acid by various particulate and soluble stimuli. Furthermore, we have shown that intravascular macrophages predominantly utilize the lipoxygenase rather than cyclooxygenase pathways to metabolize endogenous arachidonic acid. PMID- 2552226 TI - A comparison of biotin- and 35S-based in situ hybridization methodologies for detection of human papillomavirus DNA. AB - In situ hybridization is commonly used for the detection of human papillomavirus (HPV) DNA in genital tract lesions. Systems based on biotin complexes are quicker and easier to use than 35S-based systems, although reportedly less sensitive. We compared three in situ hybridization systems for HPV DNA detection: two biotin [PathoGene DNA probe assay [Enzo Diagnostics] and Viratype in situ HPV probes and HPV tissue hybridization kit [Life Technologies Inc. (LTI)] and one 35S based. By using serial sections from 80 female genital tract lesions with the histologic features of an HPV infection, sequences homologous to HPV DNA were detected in 59 cases (74%) with the LTI system and 25 cases (31%) with the Enzo system. The Enzo system uses a streptavidinbiotinylated horseradish peroxidase complex and 2% 3 amino-9-ethylcarbazole as the chromogen. The LTI system uses a streptavidin alkaline phosphatase conjugate in which the chromogen is 5-bromo-4-chloro-3 indolylphosphate in the presence of nitroblue tetrazolium. Replacing the Enzo detection system with the LTI detection system increased the sensitivity of the Enzo kit. The LTI biotin system was equally sensitive when compared against 35S labeled HPV probes. The sensitivity with the biotin probes, reported to be less than the 35S probes in a previous study (Lab Invest 58:354, 1988), was increased if the current LTI detection system replaced the detection system used in that study. It is concluded that biotin-labeled DNA probes can be equally sensitive to 35S-labeled DNA probes for the detection of sequences homologous to HPV DNA. The enhanced sensitivity for the biotin system is primarily due to improved detection of the probe/target complex. PMID- 2552227 TI - The role of prophylactic cranial irradiation in regionally advanced non-small cell lung cancer. A Southwest Oncology Group Study. AB - Lung cancer is the most common malignant disease in the United States. Only the few tumors detected very early are curable, but there has been some progress in the management of more advanced non-small cell lung cancer, particularly in regionally inoperable disease. Prevention of central nervous system relapse is an important issue in this group of patients because brain metastases ultimately develop in 20% to 25% of them. Seventy-three patients with regionally advanced non-small cell lung cancer were entered into a Phase II trial of neutron chest radiotherapy sandwiched between four cycles of chemotherapy including cisplatin, vinblastine, and mitomycin C. Prophylactic cranial irradiation was administered concurrently with chest radiotherapy (3000 cGy in 10 fractions in 15 patients; 3600 cGy in 18 fractions in the remaining 50 patients). Patients underwent computed tomographic scan of the brain before treatment and every 3 months after treatment. The initial overall response rate was 79%, but 65 of the 73 patients have subsequently died of recurrent disease. Median follow-up is 9 months for all 73 patients and 26 months for eight long-term survivors. No patient who completed the prophylactic cranial irradiation program had clinical or radiologic brain metastases. Toxic reactions to prophylactic cranial irradiation included reversible alopecia in all patients, progressive dementia in one patient, and possible optic neuritis in one patient. Both of these patients received 300 cGy per fraction of irradiation. The use of prophylactic cranial irradiation has been controversial, but its safety and efficacy in this trial supports its application in a group of patients at high risk for central nervous system relapse. Further evaluation of prophylactic cranial irradiation in clinical trials for regionally advanced non-small cell lung cancer is warranted. PMID- 2552228 TI - Induction of aplastic anemia by intra-bone marrow inoculation of a molecularly cloned feline retrovirus. AB - Intra-bone marrow inoculation of cells infected with molecularly cloned feline retrovirus (FeLV-C-Sarma [FSC]) associated with aplastic anemia was examined to test the hypothesis that cell-to-cell transmission of virus might facilitate marrow cell infection and anemogenesis, a possibility suggested by in-vitro co culture experiments. IBM inoculation of either FSC-infected feline marrow cells or fibroblasts of weanling cats bypassed age-related restriction of FSC replication, initiated viremia, caused irreversible depletion of erythroid burst forming units, and induced rapid fatal aplastic anemia. A second significant finding observed with FSC infection was pronounced systemic lymphoid depletion. The direct bone marrow inoculation system described facilitates experimental study of retrovirus-target cell interactions involved in erythroid aplasia. PMID- 2552229 TI - Inhibition of ciliary activity by phorbol esters in rabbit tracheal epithelial cells. AB - To study the effect of protein kinase C activation on respiratory ciliary activity, we measured ciliary beat frequency (CBF) by a photoelectric technique in response to phorbol esters and cell-permeable diglyceride in cultured tracheal epithelial cells from rabbits. Phorbol 12-myristate 13-acetate (PMA) resulted in a concentration- and time-dependent inhibition of CBF (half maximum inhibitory concentration (IC50) = 3 x 10(-10) M) with the maximal decrease being 21.0 +/- 1.4% (mean +/- SE, p less than 0.001) observed at 10(-6) M. L-alpha dioctanoylglycerol (DiC8), another known activator of protein kinase C, likewise reduced CBF in a dose-dependent fashion. In contrast, phorbol 12,13-didecanoate, a non-tumor-promoting phorbol ester that does not stimulate protein kinase C, produced no significant changes in CBF. The decrease in CBF induced by PMA was not affected by blockade of arachidonic acid metabolism with indomethacin and nordihydroguaiaretic acid, but was antagonized by pretreatment with H-7, a specific inhibitor of protein kinase C (p less than 0.01). Maximal ciliary inhibition with either PMA or DiC8 was not accompanied by a decrease in intracellular concentration of cyclic AMP. These results indicate that activation of protein kinase C has a significant depressive effect on ciliary activity, and hence the airway mucociliary transport function, presumably through a regulatory pathway that is not dependent on cyclic AMP or arachidonic acid metabolites. PMID- 2552231 TI - Caloric restriction perturbs the pituitary-ovarian axis and inhibits mouse mammary tumor virus production in a high-spontaneous-mammary-tumor-incidence mouse strain (C3H/SHN). AB - Dietary restriction (DR) retards aging and extends maximum life span. It is also known to decrease the incidence of hormone-dependent tumors. In the present investigation, we focussed primarily on the influence of DR on the pituitary ovarian axis, and subsequently on gene expression of the mouse mammary tumor virus. F1 females from the mating of SHN female and C3H male mice were used in this study, since these hybrids display a very high incidence of mouse mammary tumors. The mice weaned at 3 weeks were raised on either a calorically-restricted diet (DR: 50 kcal/week; N = 5) or on a control diet (C: 95 kcal/week N = 5) for 5 weeks. Three C57BL/6J Jcl ad libitum-fed female mice, 8 weeks of age, were used as reference animals since this strain has a very low incidence of mammary tumors. The mean cellular contents of prolactin (PRL) and growth hormone (GH) in the pituitary, as determined by immunohistochemistry, were found to be reduced in mice raised on the DR diet. The decrease in the mean cellular content of PRL (50% of the mean control value) was accompanied by a decrease in the number of lactotrophs (17% of the mean value of control diet mice). However, the decrease in cellular content of GH (53% of the mean control value) was not accompanied by a decrease in the number of somatotrophs (no. of somatotrophs in DR = 103% of mean control value). Histologically, ovaries from DR mice showed many growing and atretic follicles, with few corpora lutea. In contrast, both control-diet and reference (C57BL/6J Jcl) mice showed two or three corpora lutea per ovarian section. In accordance with this finding, DR mice had not established stable estrus cycles by 8 weeks of age, in contrast to both control-diet and reference mice. Since caloric restriction has been shown to decrease mammary tumor virus (MMTV) gene expression, MMTV production was investigated by electron microscopy to confirm the validity of our experimental conditions. In DR or reference C57BL/6J Jcl mice, MMTV particles were rarely found in the mammary gland samples, but were always found in samples from control mice. The development of mammary glands, as indicated by the number of villi or the development of the rough endoplasmic reticulum, was delayed in DR mice. Thus, it was concluded that caloric restriction decreases the number of lactrotrophs, inhibits ovulation and delays mammary gland development. This immature status is considered to be due to perturbations in the pituitary-ovarian axis by caloric restriction. PMID- 2552230 TI - Superoxide anion radical production in different animal species. AB - The general objective of this study was to examine the relationship between oxygen free radicals and the aging process. The rate of superoxide anion radical (O2.-) generation was measured in liver sub-mitochondrial particles from mouse, rat, rabbit, pig and cow, and in flight muscle sub-mitochondrial particles from the housefly. The rate of O2.- generation was determined as superoxide dismutase inhibitable reduction of ferricytochrome c in the presence of antimycin A and KCN. O2.- generation was inversely related to maximum species life span potential (MLSP) (r = -0.92). A 24-fold difference in the rate of O2.- production was observed between the cow and the fly while a 6-fold difference existed among the mammals. The results are interpreted to indicate that under identical conditions, mitochondria from organisms with low MLSP have a relatively greater propensity to generate O2.-. This may be suggestive of innate differences in the molecular organization of the inner mitochondrial membrane among different species. PMID- 2552232 TI - [Delayed polyneuropathy induced by organophosphorus compounds: a great unknown]. PMID- 2552233 TI - [Application of electron microscopy in the rapid diagnosis of congenital cytomegalovirus infections]. PMID- 2552234 TI - [Acute idiopathic pleuropericarditis associated with transitory hypoparathyroidism]. PMID- 2552235 TI - [Myositis with inclusion bodies: a little-known variety of idiopathic inflammatory myopathy]. AB - The authors report two male patients with inclusion body myositis (IBM) and review the features of this condition in the literature. This is an uncommon type of idiopathic myositis which involves males more often than females; it usually develops in elderly patients; its course is very slow; it is usually associated with distal weakness; neurophysiological studies show a mixed "myogenic" and "neurogenic" pattern; and it is usually unresponsive to corticosteroids. The diagnosis is basically made on the basis of the histological features, mainly consisting of vacuoles surrounded by a basophilic haze in histochemical stains of frozen tissue and, particularly, by the presence of characteristic microfilaments in ultrastructural studies. PMID- 2552236 TI - [Myositis with inclusion bodies]. PMID- 2552237 TI - [Immunologically disturbed energy and calcium homeostasis as a possible pathogenesis of dilated cardiomyopathy]. PMID- 2552238 TI - Differences in the ability of cells to fuse are mediated by strains of Epstein Barr virus. AB - Epstein-Barr virus (EBV) preparations from both NPC-KT cells (NPC-EBV) and P3HR-1 cells (HR-1-EBV) can induce cell fusion between EBV receptor (EBVR)-positive Raji cells and EBVR-negative cells, but other strains of EBV cannot induce cell fusion. The effect of these two EBV isolates on ability of cells to fuse has been studied to determine if there are differences in the biological properties of the different EBV isolates, particularly the isolates obtained from nasopharyngeal carcinoma such as NPC-EBV. The frequency of cell fusion between NPC-EBV superinfected Raji cells and EBVR-negative epithelial cells (Ad-AH) was increased more than 30-fold in the presence of medium containing 1% dimethylsulfoxide (DMSO). However, the frequency of cell fusion between HR-1-EBV-superinfected Raji cells and Ad-AH cells was unaffected under the same conditions. The data show that differences in the ability of cells to fuse are induced by variants of EBV in response to DMSO. These differences may be important in elucidating the different biological properties of EBV isolates and might have implications for the pathophysiology of EBV-associated illness. PMID- 2552239 TI - Malignant transformation of a nasopharyngeal angiofibroma. AB - The malignant transformation of a nasopharyngeal angiofibroma is a rare occurrence. This report describes the development of a fibrosarcoma in a patient over a 20-year period after four surgical procedures and two courses of radiation therapy to control a nasopharyngeal angiofibroma. A review of the literature revealed similar descriptions of this problem. Radiation likely plays a major role in causing the malignant change. An initial complete surgical excision can prevent recurrence of a nasopharyngeal angiofibroma as well as sarcomatous transformation. With the availability of surgical procedures that can completely and safely resect almost all nasopharyngeal angiofibromas, radiation therapy can be avoided. PMID- 2552240 TI - Endothelin stimulates release of atrial natriuretic peptides in vitro and in vivo. AB - The effect of endothelin (END) on the release of atrial natriuretic peptides (ANP) was studied in isolated rat atria and in conscious rats. END stimulates the ANP release in vitro in a dose-dependent manner. An increase in ANP plasma levels and cyclic GMP plasma levels was also observed in conscious rats after injection of END. When a monoclonal antibody directed against ANP was injected together with END the increase in cyclic GMP was completely blocked. From this study it is concluded that END is a potent secretagogue for ANP both in vitro and in vivo. PMID- 2552241 TI - A high affinity, highly selective ligand for the delta opioid receptor: [3H]-[D Pen2, pCl-Phe4, d-Pen5]enkephalin. AB - Binding characteristics of a new, conformationally constrained, halogenated enkephalin analogue, [3H]-[D-penicillamine2, pCl-Phe4, D penicillamine5]enkephalin ([3H]pCl-DPDPE), were determined using homogenized rat brain tissue. Saturation binding studies at 25 degrees C determined a dissociation constant (Kd) of 328 +/- 27.pM and a receptor density (Bmax) of 87.2 +/- 4.2 fmol/mg protein. Kinetic studies demonstrated biphasic association for [3H]pCl-DPDPE, with association rate constants of 5.05 x 10(8) +/- 2.5 x 10(8) and 0.147 +/- 10(8) +/- 0.014 x 10(8) M-1 min-1. Dissociation was monophasic with a dissociation rate constant of 2.96 x 10(-3) +/- 0.25 x 10(-3) min-1. The average Kd values determined by these kinetic studies were 8.4 +/- 2.7 pM and 201 +/- 4 pM. Competitive inhibition studies demonstrated that [3H]pCl-DPDPE has excellent selectively for the delta opioid receptor. [3H]pCl-DPDPE binding was inhibited by low concentrations of ligands selective for delta opioid receptor relative to the concentrations required by ligands selective for mu and kappa sites. These data show that [3H]pCl-DPDPE is a highly selective, high affinity ligand which should be useful in characterizing the delta opioid receptor. PMID- 2552242 TI - Photoaffinity labeling of [3H]flunitrazepam- and [3H]Ro15-4513-bound pellets in rat cerebral cortex and cerebellum. AB - Irreversible incorporation of [3H]flunitrazepam and [3H]Ro15-4513 into GABA/benzodiazepine receptor subunits was studied by UV irradiation using ligand bound membrane pellets from rat cerebral cortical and cerebellar synaptic membranes. Specific incorporation for [3H]flunitrazepam was greater in the pellet than in the suspension. The incorporation was identical for [3H]Ro15-4513 in both pellet and suspension. With the ligand-bound pellets, 50% of the available binding sites were photolabeled by both ligands in cortex and cerebellum. SDS polyacrylamide gel electrophoresis and fluorography of [3H]flunitrazepam photo labeled receptor revealed the same number of major sites in both brain regions. In contrast, [3H]Ro15-4513 appears to label fewer sites in cortex and cerebellum. Photoaffinity labeling with [3H]flunitrazepam in ligand-bound membrane pellet provides a more selective and reliable method for studying the subunit structure of GABA/benzodiazepine receptor complex. PMID- 2552243 TI - Inhibition of polyphosphoinositide turnover in rat cerebral cortex by clonidine. AB - In the rat brain, a number of receptors are linked to phospholipase C which catalyzes the hydrolysis of membrane inositol phospholipids; stimulation of alpha 1-adrenergic receptors, for example, increases polyphosphoinositide turnover, but stimulation of alpha 2-receptors does not. The hydrolysis of inositol phospholipids in rat cortical slices was investigated using a direct assay involving prelabeling these lipids with 3H-inositol and then measuring the formation of 3H-inositol phosphates in the presence of lithium ions. As expected, clonidine, an alpha 2-agonist, did not stimulate the formation of 3H-inositol phosphates; however, clonidine antagonized the ability of noradrenaline to stimulate 3H-inositol phosphate formation. This effect was not blocked by antagonists of alpha 2, 5HT2, H2, or muscarinic receptors. Clonidine did not affect carbachol-stimulated 3H-inositol phosphate formation. PMID- 2552244 TI - Immunoneutralization of corticotropin-releasing hormone prevents the diurnal surge of ACTH. AB - To determine whether CRH is required for the evening rise in plasma ACTH, rats were injected at 0800 hr with CRH antiserum (anti-CRH) or normal rabbit serum (NRS). Blood samples were taken through venous catheters at 0800 hr before treatment and at 1300, 1700, and 2100 hr. Plasma was assayed for immunoreactive ACTH and corticosterone. There was no significant difference in pretreatment values between the two groups. Immunoneutralization of CRH abolished the rise in plasma ACTH seen at 1700 hr in the NRS group but had little effect on earlier levels. The diurnal elevation in plasma corticosterone continued after anti-CRH treatment, but peak levels occurred earlier. Plasma ACTH and corticosterone were significantly correlated at the time of the diurnal surge, but not at 0800 hr or 1300 hr in the NRS controls or at any time point in the anti-CRH group. These results suggest that CRH is required for the diurnal surge of plasma ACTH. They also confirm previous observations by others that the adrenal cortex does not require active CRH or a diurnal surge of ACTH in order to exhibit a significant diurnal increase in secretion of corticosterone, and that factors other than CRH may be relatively more active than CRH in regulation of ACTH secretion during the time of circadian inactivity. PMID- 2552245 TI - Effects of synthetic and naturally occurring flavonoids on Na+,K+-ATPase: aspects of the structure-activity relationship and action mechanism. AB - A comparative study was made of the effects of 15 synthetic and naturally occurring flavonoids on the hydrolytic activity of Na+, K+-adenosine triphosphatase (ATPase). Twelve of the flavonoids examined were mono-hydroxy or mono-methoxy derivatives. All inhibited Na+, K+-ATPase from dog kidney cortex when present at concentrations from 40-1000 microM. Flavones possessing cyclohexyl instead of the phenyl group (i.e., 2-cyclohexyl-benzopyran-4-one derivatives), were the most potent with IC50 at 257-320 microM. Structure activity relationships were observed among the following mono-substituted flavones as: i) 2-cyclohexyl-benzopyran-4-one much greater than 2-phenyl benzopyran-4-one; ii) 2-cyclohexyl-7-hydroxybenzopyran-4-one greater than 2 cyclohexyl-6-hydroxybenzopyran-4-one greater than 2-cyclohexyl-5 hydroxybenzopyran-4-one. Some flavonoids showing potent inhibitory activity were also examined for ouabain-displacement activity on human erythrocytes. Hardly any of the flavonoids were able to block [3H]ouabain binding to erythrocytes. These results suggest that the mechanism by which flavonoid block Na+, K+-ATPase is not related to the cardiac glycoside-specific binding site(s) of this enzyme. PMID- 2552246 TI - Melanin concentrating hormone. I. Influence of nerves and hormones on the control of trout melanophores. AB - When melanophores on trout scales are cultured in vitro they show a transitory melanin concentration, which can be prevented by addition to the medium of the alpha adrenergic blocker, phentolamine. This indicates the release of endogenous nor-epinephrine from local nerve terminals. This initial phase of melanin aggregation is followed by redispersion and then by a second, more gradual melanin concentration over several days, which is not antagonized by phentolamine. A final melanophore index of between 2-2.5 is attained which may be the resting state of trout melanophores. Using short-term cultured melanophores which have passed the phase of endogenous nor-epinephrine release, it is shown that exogenous nor-epinephrine will interact synergistically with the melanin concentrating hormone to achieve full melanin concentration. Evidence is discussed for believing that in the trout, such synergy is necessary to achieve maximum pallor in vivo. PMID- 2552247 TI - Melanin concentrating hormone. V. Isolation and characterization of alpha melanocyte-stimulating hormone from frog pituitary glands. AB - The structure of alpha-melanocyte-stimulating hormone (alpha-MSH) has been determined in the pars intermedia of the frog Rana ridibunda. Pulse-chase labeling of frog neurointermediate lobes with selective amino acids revealed that the composition of frog alpha-MSH is similar to that of alpha-MSH from all mammalian species yet studied. Tryptic mapping of nexly synthetized alpha-MSH generated two fragments with the following amino acid composition: (T1) Trp, Pro, Lys, Gly, Val and (T2) Tyr, Arg, Phe, His, Ser, Glu. Concurrently, alpha-MSH was purified from 100 neurointermediate lobes to apparent homogeneity by reverse phase HPLC. The sequence of the peptide determined by automated Edman degradation was Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Lys-Pro-Val. The structure of frog alpha-MSH is thus identical to mammalian des-N alpha-acetyl alpha-MSH and differs from the sequence of toad (Xenopus laevis) alpha-MSH only by the first residue (Ser instead of Ala). These results confirm that the sequence of alpha-MSH has been highly preserved during evolution. PMID- 2552248 TI - Presence and release of calcitonin gene-related peptide in rat stomach. AB - Immunoreactive (IR)-calcitonin gene-related peptide (CGRP) was identified throughout the entire stomach of rats, being most highly concentrated in the pyloric region, and the concentrations in muscular layers being higher than those in mucosal layers. In addition, IR-CGRP was also present in the venous effluent from isolated perfused rat stomach, and its release was stimulated by dibutyryl cyclic AMP or theophylline but not by glucagon. Gel chromatography as well as HPLC of both tissue extracts and gastric perfusate showed three identical major peaks of IR-CGRP, one of which coeluted with synthetic CGRP. These results suggest that CGRP in the stomach plays a role in the regulation of gastric function. PMID- 2552249 TI - The high cost and low efficacy of weekly viral cultures for pregnant women with recurrent genital herpes: a reappraisal. AB - The use of weekly cultures to prevent neonatal infection among infants of pregnant women who have histories of genital herpes has been controversial since a decision analysis study in 1983 suggested that this strategy was not cost effective and would avert relatively few cases of neonatal herpes simplex virus infection. Using more recent and better data, the authors reanalyzed this approach to reducing neonatal herpes infection. The reanalysis revealed that a national screening program would prevent only 1.8 cases of neonatal herpes in the United States annually, at a cost of more than 37 million dollars per case averted. The program would cost nearly 7 million dollars per quality-adjusted life year gained when only infant deaths are taken into consideration. When maternal deaths from excess cesarean deliveries are taken into account, over 44 million dollars would be spent for every quality-adjusted life-year gained. On the basis of the strategy's limited benefits and low cost-effectiveness, the authors support the American College of Obstetrics and Gynecology's position of abandoning the recommendation for weekly prenatal herpes cultures. PMID- 2552250 TI - Mediastinal staging in non-small-cell lung cancer. AB - Decision analysis was used to study the approach to staging mediastinal involvement in patients with non-small-cell lung cancer (NSCLC). Various diagnostic strategies for mediastinal disease staging using computed tomography (CT), mediastinoscopy, and bronchoscopy with transbronchial needle aspiration (TBNA), either individually or in series, were compared and found to result in similar patient life expectancies. Two strategies, one using bronchoscopy and TBNA alone and the other using it in combination with CT, were consistently least expensive across a wide range of prior probabilities, test characteristics, and charges. The authors conclude that strategies for staging mediastinal involvement in NSCLC that rely on bronchoscopy and TBNA are preferable because they are least expensive. PMID- 2552251 TI - Comparative functional study of Dexon and nylon microsutures in rat uterine anastomoses. AB - Forty-two female Wistar rats were used in a long-term study designed to evaluate the functional sequelae of employing 10-0 Dexon "S" and nylon microsutures in uterine anastomoses. Functional criteria used were the number of fetuses and live births, during and after pregnancy, respectively. Results indicate that the rats from both control and experimental groups were likely to have identical litter sizes for the first 3 deliveries (P greater than 0.01). An unexplained observation was the finding of a larger litter size for the experimental group rats at the fourth delivery as compared to its control counterpart (P less than 0.01). Also explicit is the propensity for the right uterine horn to bear a larger number of fetuses than its counterpart (P less than 0.01). Finally, neither suture was associated with a larger number of fetuses (P greater than 0.01). It was concluded that 1) uterine anastomosis did not prove detrimental to the functional outcome, 2) none of the sutures proved functionally superior, and 3) nylon was more likely to be associated with foreign body and inflammatory responses. PMID- 2552252 TI - Concerted evolution of light satellite DNA in genus Mus implies amplification and homogenization of large blocks of repeats. AB - Light satellite DNA components present in species belonging to the genus Mus and to related murids were studied using the Southern blot technique. The results show species variations in both the amount and periodic structure of the repeating units, which suggests that families of related higher-order repeats developed in a common ancestor and were then amplified and/or deleted to different extents during the subsequent evolutionary period. Although the patterns generated by a series of type B enzymes (restriction enzymes that possess sites in a limited number of segments making up the total satellite DNA) in the species closely related to the M. musculus complex were very similar, sequence analysis of cloned unit repeats in two of these species (M. musculus domesticus and M. spretoides) showed near fixation of species-diagnostic variant nucleotides. This suggests that the important amplification and homogenization events that occurred after the divergence of M. spretus must have involved large blocks of sequences. PMID- 2552253 TI - Restriction-map variation with the yellow-achaete-scute region in five populations of Drosophila melanogaster. AB - It has been proposed that the degree of recombination for a genomic region will affect the level of both nucleotide heterozygosity and the density of transposable elements. Both features of genomic diversity have been examined in a number of recent reports for regions undergoing relatively normal levels of recombination in Drosophila melanogaster. In this study the genomic variation associated with yellow-achaete-scute loci located at the tip of the X chromosome is examined by six-cutter restriction mapping. In this region, as usual for regions adjacent to telomeres, crossing-over is dramatically reduced, and published studies of visible mutants indicate extremely little restriction-map variation. Eight six-cutter restriction endonucleases were used to locate sequence variation in 14- and 16.5-kb regions in 109 lines sampled from North America, Africa, and Europe. The overall level of heterozygosity is estimated as 0.29%. Nine large insertions, all presumed to be transposable elements, were observed. Base-pair heterozygosity appears to be reduced compared with regions having normal levels of recombination. The estimated heterozygosity is much higher than reported in earlier studies of restriction-map variation among visible mutations in the complex. The incidence of large insertions is not elevated compared with that in other regions of the genome. This suggests that asymmetric synapsis and exchange is not an important mechanism for the elimination of transposable elements. PMID- 2552254 TI - Satellite DNA and higher-primate phylogeny. PMID- 2552255 TI - Characterization of Rhizobium phaseoli Sym plasmid regions involved in nodule morphogenesis and host-range specificity. AB - Two nodulation regions from the symbiotic plasmid (pSym) of Rhizobium phaseoli CE 3 were identified. The two regions were contained in overlapping cosmids pSM927 and pSM991. These cosmids, in a R. phaseoli pSym-cured strain background, induced ineffective nodules on Phaseolus vulgaris roots. Transconjugants of Rhizobium meliloti harbouring pSM991 induced nodule-like structures on bean roots, suggesting that this cosmid contains host-range determinants. Analysis of deletions and insertional mutations in the sequences of pSM991 indicated that the genes responsible for the induction and development of nodules in P. vulgaris are organized in two regions 20 kb apart. One region, located in a 6.8 kb EcoRI fragment, includes the common nodABC genes. The other region, located in a 3.5 kb EcoRI fragment, contains information required for host-range determination. PMID- 2552256 TI - The nifA gene product from Rhizobium leguminosarum biovar trifolii lacks the N terminal domain found in other NifA proteins. AB - The nifA gene has been identified between the fixX and nifB genes in the clover microsymbiont Rhizobium leguminosarum biovar trifolii (R.I. bv. trifolii) strain ANU843. Expression of the nifA gene is induced in the symbiotic state and site directed mutagenesis experiments indicate that nifA expression is essential for symbiotic nitrogen fixation. Interestingly, the predicted R.I. bv. trifolii NifA protein lacks an N-terminal domain that is present in the homologous proteins from R.I. bv. viciae, Rhizobium meliloti, Bradyrhizobium japonicum, Klebsiella pneumoniae and all other documented NifA proteins. This indicates that this N terminal domain is not essential for NifA function in R.I. bv. trifolii. PMID- 2552257 TI - A Mycoplasma genetic element resembling prokaryotic insertion sequences. AB - Nucleotide sequence analysis of two Mycoplasma hyopneumoniae-derived copies of a repetitive genetic element revealed structural similarities to typical prokaryotic insertion sequences. This is the first such sequence identified in the class Mollicutes. The element spans approximately 1550bp, with 28bp inverted terminal repeats. Two open reading frames occur within the sequence, one potentially encoding a protein with a size-variant alpha-helical domain containing heptameric leucine periodicity. Hybridization data with several strains from each of two mycoplasma species showed that the repetitive sequence is variably distributed within the M. hyopneumoniae and Mycoplasma hyorhinis chromosomes and indicated that in some cases the repeated sequence is contained within a larger genetic element which may be the result of phage or plasmid insertion. PMID- 2552258 TI - Specificity of insertion of IS91, an insertion sequence present in alpha haemolysin plasmids of Escherichia coli. AB - We have determined the DNA sequences of eight different insertions of IS91 in a specifically engineered recipient plasmid of known DNA sequence (pSU300). The sequences at the termini of IS91 are 5'-CGAGTAGG...CCTATCGAT. IS91 inserts specifically 5' to either one of the tetranucleotides 5'-GAAC or 5'-CAAG, and always in the same relative orientation with respect to the sequence of the target. Except in one special case, no duplications of the recipient DNA were produced at the site of insertion. PMID- 2552259 TI - Analysis of the chromosomal location of two copies of a Bordetella pertussis insertion sequence. AB - IS481v1 and IS481v2 are two copies of a Bordetella pertussis insertion sequence element. We have shown that IS481v1 is located within 3 kbp of the start of the adenylate cyclase gene whilst IS481v2 is immediately adjacent to the end of the agglutinogen 2 gene and provides the stop codon for that gene. In addition, IS481v1 and IS481v2 were present at these two specific sites in nine strains of B. pertussis, including two Phase IV strains which expressed neither adenylate cyclase nor agglutinogen 2 and three Phase I strains which did not express agglutinogen 2. The loss of expression in these strains is not the result of DNA rearrangements at the sites of IS481v1 or IS481v2. PMID- 2552260 TI - Characterization of amber mutations in bacteriophage Mu transposase: a functional analysis of the protein. AB - We have characterized a series of amber mutations in the A gene of bacteriophage Mu encoding the phage transposase. We tested different activities of these mutant proteins either in a sup0 strain or in different sup bacteria. In conjunction with the results described in the accompanying paper by Betermier et al. (1989) we find that the C-terminus of the protein is not absolutely essential for global transposase function, but is essential for phage growth. Specific binding to Mu ends is defined by a more central domain. Our results also reinforce the previous findings (Betermier et al., 1987) that more than one protein may be specified by the A gene. PMID- 2552261 TI - Functional domains of bacteriophage Mu transposase: properties of C-terminal deletions. AB - We have generated a series of 3' deletions of a cloned copy of the bacteriophage Mu transposase (A) gene. The corresponding truncated proteins, expressed under the control of the lambda PI promoter, were analysed in vivo for their capacity to complement a super-infecting MuAam phage, both for lytic growth and lysogeny, and for their effect on growth of wild-type Mu following infection or induction of a lysogen. Using crude cell extracts, we have also examined binding properties of these proteins to the ends of Mu. The results allow us to further define regions of the protein important in replicative transposition, establishment of lysogeny and DNA binding. PMID- 2552262 TI - Molecular cloning and expression of a locus (mdoA) implicated in the biosynthesis of membrane-derived oligosaccharides in Escherichia coli. AB - Mutants of Escherichia coli defective in the mdoA locus are blocked at an early stage in the biosynthesis of membrane-derived oligosaccharides. The mdoA locus has now been cloned into multicopy plasmids. A 5 kb DNA fragment is necessary to complement mdoA mutations. Cells harbouring the mdoA+ plasmid produced three to four times more MDO than wild-type cells. MDO overproduction did not affect the degree of MDO substitution with sn-1-phosphoglycerol residues. The biosynthesis of MDO remained under osmotic control in overproducing strains. PMID- 2552263 TI - Cloning a genomic region required for a high-affinity iron-uptake system in Rhizobium meliloti 1021. AB - A collection of transposon-induced mutants of Rhizobium meliloti 1021 defective in siderophore-mediated iron assimilation were obtained and classified as biosynthetic, transport or regulatory. Several of the mutations were cloned and the adjacent sequences were used to acquire complementing DNA from the wild type. A single genomic region of about 35kb complemented all of the mutants deficient in production of the siderophore. PMID- 2552265 TI - Osmotic regulation of porin expression: a role for DNA supercoiling. AB - The OmpC and OmpF porins are major outer membrane proteins of Escherichia coli and Salmonella typhimurium. Their expression is affected by many environmental factors and by mutations in a variety of independent genes. The pair of regulatory proteins, OmpR and EnvZ, are required for normal porin expression. Despite intensive investigation, the mechanisms by which porin expression is regulated remain unclear. Mutations which alter supercoiling, as well as inhibitors of DNA gyrase, show that porin expression is extremely and specifically sensitive to the level of DNA supercoiling. Our data lead us to suggest that environmentally induced changes in DNA supercoiling may play a role in determining the level of porin expression. These findings have implications for current models of porin regulation. PMID- 2552264 TI - Functional organization and nucleotide sequence of virulence Region-2 on the large virulence plasmid in Shigella flexneri 2a. AB - The 7 kb virulence Region-2 of the large (virulence) plasmid in Shigella flexneri 2a encodes several proteins required for invasion of intestinal epithelial cells. Insertion and deletion mutagenesis, DNA subcloning and SDS-polyacrylamide gel electrophoresis of proteins synthesized in minicells demonstrated five genes in this region. They encode 24, 18, 62 (IpaB), 41 (IpaC) and 37 (IpaD)-kiloDalton (kD) proteins. Complementation of Tn5-induced mutations in Region-2 with the above plasmid constructs indicated that Region-2 consists of two operons and that the three Ipa proteins are essential for the virulence phenotype. The transcriptional organization determined by Northern blotting, S1 nuclease protection and the effect of Tn5 insertions on expression of the Ipa proteins revealed that Region-2 has three promoters that transcribe RNAs of 4.0, 4.5 and 7.5 kb. The 4.0 kb RNA was the transcript for the operon encoding the 24, 18 kD, IpaB and C proteins and the 4.5 kb RNA for the ipaD gene. In addition, the full length RNA of 7.5 kb which covers Region-2 supplemented full expression of the Ipa proteins. The 7663 nucleotides of Region-2 were determined to confirm the five open reading frames encoding 23,655, 17,755, 62,168, 41,077 and 36,660 Dalton proteins, respectively, and their regulatory sequences. PMID- 2552267 TI - Free radicals in stouts and ales. PMID- 2552266 TI - Unprocessed insulin proreceptor in cultured fibroblasts from a patient with extreme insulin resistance. AB - Insulin receptors and IGF-I receptors in cultured fibroblasts were investigated in a patient with extreme insulin resistance due to unprocessed insulin receptors. Insulin binding to cultured fibroblast monolayers and partially purified insulin receptors was extremely decreased to 27% and 18% of control value, respectively. Affinity cross-linking study revealed that molecular weight of the insulin receptor was 210 kDa and that it could not be dissociated to alpha and beta-subunit with dithiothreitol treatment. Because IGF-I binding to the fibroblasts from the patient was normal and alpha-subunit of IGF-I receptor was 135 KDa, the defect was specific to the insulin receptor. Autophosphorylation of the 210 kDa unprocessed insulin proreceptor was stimulated by insulin in a dose dependent manner. In the fibroblasts from the patient, insulin-stimulated alpha aminoisobutyric acid uptake was fivefold shifted to the right in the dose response curve (ED50 20 ng/mL for the patient v 3.5 ng/mL for the control subjects), but the maximally stimulated uptake was normal. With 0.025% trypsin treatment, insulin binding and alpha-aminoisobutyric acid uptake were normalized. These results suggested that (1) abnormal processing of insulin proreceptor also occurred in the cultured fibroblasts, (2) the postreceptor steps of insulin action were totally intact, and (3) IGF-I receptors were normally processed in this patient. PMID- 2552268 TI - Metalloproteinases are not involved in the phagocytosis of collagen fibrils by fibroblasts. AB - The effect of various metalloproteinase-inhibiting compounds on collagen phagocytosis by fibroblasts was studied in cultured periosteal tissue. Evidence is presented indicating that neither anti-collagenase nor anti-stromelysin interfere with the uptake of collagen fibrils from the extracellular space and their intracellular digestion. Similar results were obtained with tissue inhibitor of metalloproteinases (TIMP). In the presence of the proteinase inhibitor leupeptin, a compound which strongly inhibits the intracellular degradation of phagocytosed collagen, a time-dependent increase in the amount of internalized collagen was found. This increase proved to be similar in explants treated as well as in those not treated with the metalloproteinase-inhibiting compounds. It is concluded that enzymes, such as collagenase and stromelysin, do not play a crucial role in the phagocytosis and intracellular digestion of collagen fibrils by fibroblasts. If these enzymes are involved it must be prior to these events. Based on the morphometric data the intralysosomal degradation time of collagen was calculated to be about 30 minutes. A comparison with findings in the literature on collagen metabolism in the periodontal ligament of the rat molar suggests that all collagen degraded may pass through the phagolysome pathway during physiological turnover and remodelling. PMID- 2552269 TI - Human skin and post-burn scar hyaluronan: demonstration of the association with collagen and other proteins. AB - Hyaluronan (HA) extracted from tissues has been demonstrated to have an enhancing effect on the process of wound healing; the question arises whether this effect is due to the HA or to associated collagen and other proteins. In this study, HA has been extracted from human skin and scar tissue under dissociative conditions and isolated by DEAE-cellulose chromatography followed by CsCl gradient and Sepharose CL-6B chromatography. This highly purified HA was found to contain between 4 and 28% protein, with collagen constituting 5% of the total protein. A functional association between HA and a collagen protein complex is proposed. PMID- 2552270 TI - Cortical benzodiazepine receptor binding in a rabbit model of hepatic encephalopathy: the effect of Triton X-100 on receptor solubilization. AB - Increased benzodiazepine (BZ) receptor density has been reported in brains of rabbits with hepatic encephalopathy (HE) due to galactosamine (GalN)-induced fulminant hepatic failure (FHF). These data were generated using detergent-Triton X-100-treated neural membranes. While performing further studies it was noted that the increase in BZ receptor density was not demonstrable when Triton X-100 preparation was not employed. Accordingly the binding of [3H] flunitrazepam, a BZ ligand, to neural membranes from cortices of normal rabbits and rabbits with HE due to (GalN)-induced FHF was studied with and without detergent preparation. Scatchard plot analysis of the binding data indicated that when no detergent was employed, the apparent affinity and density of BZ receptors were similar for control membranes and membranes from animals in HE. BZ receptors from animals in HE were shown to be more resistant to solubilization by Triton than control membranes. These findings (a) afford a potential explanation for the apparent increase in density of BZ receptors in this model when Triton treatment of neural membranes is utilized and (b) suggest that recent evidence for increased GABAergic tone in the syndrome of HE is not dependent on an increased density of BZ receptors. PMID- 2552272 TI - Receptor-mediated Ca++ entry in blood vessels. AB - The importance of receptor-mediated Ca++ entry (RMCa++E) relative to Ca++ release and potential-dependent Ca++ entry (PDCa++E) in agonist-induced responses in rabbit aorta and renal artery was quantitatively delineated by utilizing a solution without added Ca++ containing low ethyleneglycol bis(beta aminoethylether)-N,N'-tetraacetic acid (EGTA) plus D600 to inhibit PDCa++E. Adding an approximate ED80 concentration of norepinephrine (NE; 3 x 10(-7) M), histamine (Hist; 3 x 10(-6) M), or serotonin (5HT; 3 x 10(-6) M) to this solution results in a transient increase in developed force that is attributed to release of a limited cellular pool of Ca++. (NE greater than Hist much greater than 5HT). When the concentrations are approximately equipotent (NE, 3 x 10(-7) M; Hist, 3 x 10(-5) M; 5HT, 10(-5) M) the Ca++ release component increases for Hist and 5HT such that NE = Hist greater than 5HT. Subsequent addition of Ca++ results in an increase in developed force that is sustained and represents RMCa++E. In aorta, RMCa++E can account for 91% of the total NE-induced developed force; for an equipotent concentration of Hist, 71%; and for an equipotent concentration of 5HT, only 37%. This capacity for stimulating RMCa++E is inversely related to the sensitivity of these agonists to the PDCa++E blocker, D600 (5HT much greater than Hist greater than NE). Chronic Mg++ potentiates control responses to NE in normal Ca++, but depresses the sensitivity to Ca++ in the RMCa++E concentration response relationship. The sustained response associated with RMCa++E is only minimally relaxed or inhibited by Mg++ (acute) and is completely inhibited or slowly and completely relaxed by La . In renal artery, a similar approximate ED80 concentration of NE (3 x 10(-6) M) results in a NE-induced transient response attributed to Ca++ release that is 60% less than that seen in aorta, whereas the RMCa++E component in renal artery accounts for 78% of the total response (only 10% less than in aorta). Thus, it appears that there are pharmacologically distinct Ca++ channels in some blood vessels that are differentially activated in a selective and potential-independent manner as a result of specific agonist receptor interactions. PMID- 2552273 TI - Effects of inorganic and organic promoters and inhibitors of calcium influx on stretch-induced myogenic tone of vascular tissues. AB - The static or dynamic stretch of helical strips or sudden increase of the transmural pressure produced contraction in cerebral and coronary arteries isolated from dogs and rabbits. The simultaneous recording of Ca transients and contractile activity showed that fluorescence intensity of fura-2 loaded cerebral artery increased during stretch activation. The contractile activation due to stretch was augmented by potentiators of Ca2+ influx, while it was attenuated by inhibitors of Ca2+ influx. The results suggest that contractile activation by mechanical stretch seems to provide a useful alternative technique to electrical and pharmacological stimuli for use in studies on contraction of vascular tissues and the effects of drugs and ions such as Ca antagonists and divalent cations which affect Ca2+ movement. PMID- 2552274 TI - The effects of prenatal exposure to delta-9-tetrahydrocannabinol on the rest activity cycle of the preweanling rat. AB - Either 15 or 50 mg/kg of delta-9-tetrahydrocannabinol (delta-9-THC) was administered from Day 2 through Day 22 of gestation. Pair-fed (0 mg/kg) and nontreated groups served as controls and all treated and control litters were fostered at birth to untreated dams. To examine whether delta-9-THC produces effects on the rest-activity cycle of the offspring, groups of 3 littermates from each of the treated and control groups were tested for an 8 hr observation period on electronic activity monitors at 17, 22 and 30 days of age. Whereas all treated and control groups showed age-dependent changes in activity, neither activity level nor the rest-activity pattern were affected by delta-9-THC. These findings are discussed in relation to neurobehavioral studies of activity changes following prenatal exposure to cannabis. PMID- 2552271 TI - Human herpesviruses: a consideration of the latent state. PMID- 2552276 TI - [Malignant fibrous histiocytoma of the gums. Anatomo-clinical study of a case]. AB - A case of malignant fibrous histiocytoma occurring in the gingival tissue on the site of the mandibular right canine is reported. The histological, histochemical, immunohistochemical and ultrastructural features are discussed and compared with the recent literature on malignant fibrous histiocytomas. PMID- 2552275 TI - Formamidine pesticides and alpha 2-adrenoceptors: studies with amitraz and chlordimeform in rats and development of a radioreceptor binding assay. AB - The interaction of the formamidine pesticides chlordimeform (CDM) and amitraz (AMZ) with rat brain alpha2-adrenoceptors was investigated. Both compounds inhibited the binding of 3H-clonidine and 3H-yohimbine in vitro with IC50 values of 62-68 microM (CDM) and 95-110 nM (AMZ). In vivo administration of AMZ and CDM caused a dose-dependent inhibition of 3H-clonidine binding in rat forebrain. The inhibition was short-lasting (24 hr) following CDM administration, while after AMZ recovery of 3H-clonidine binding occurred only after 72 hr. Good correlations were found between inhibition of brain 3H-clonidine binding by the formamidines and "plasma equivalents" of these compounds and/or their biologically active metabolites, as measured by a new radioreceptor assay. These results suggest that 1) formamidines can interact in vivo with brain alpha 2-adrenoceptors when administered at doses previously shown to cause toxic effects on the central nervous system: and 2) this effect is reversible, both in vivo and in vitro, and appears to be linked to the presence of the formamidines and/or their active metabolites at the receptor sites. PMID- 2552277 TI - Perinatal death, the family, and the role of the health professional. AB - In order to substantially assist a grieving family at the time of perinatal death, the health care team should have sufficient knowledge of the grief reaction, including the duration, maternal/paternal differences, and perinatal psychodynamics. Immediate support includes bereavement counseling. Predelivery care should be provided, if appropriate. The health care team should be prepared for common questions and know about local cultural traditions. Follow-up care after discharge, including anticipatory guidance, is important. The issues of repeat pregnancies and holiday behavior should be addressed. Siblings must be taken into account. The health care team should be able to detect unresolved grief and identify at-risk parents. A hospital protocol should be in place. It should be kept in mind that parents remember the emotional care given to them rather than the technical or medical care provided. PMID- 2552278 TI - The role of the nurse in assisting with eye examinations on premature infants. PMID- 2552279 TI - Acute leukemia after adjuvant chemotherapy for breast cancer. AB - Alkalating chemotherapeutic agents are frequently implicated in the development of acute non-lymphocytic leukemia. A case report illustrates the danger of administering a prolonged course of adjuvant chemotherapy with Melphalan following mastectomy for breast cancer. PMID- 2552280 TI - [Hydroxyproline in breast cancer]. AB - The content of hydroxyproline was determined in cancerous and noncancerous tissues obtained from 40 patients with primary breast cancer, and investigated from a clinicopathologic viewpoint. The level of hydroxyproline in cancerous tissue was higher than that in normal mammary gland, but was lower than that of the tissue of mastopathy. In relation to the histologic type, the level was the highest in scirrhous carcinoma, followed by papillotubular and solid tubular carcinoma. With regard to lymph node metastasis, the hydroxyproline level was significantly higher in positive than in negative metastatic cases. The corresponding level tended to be higher in patients at stages II-IV than in those at stage I. There was no obvious relationship to tumor size. However, a significant, negative correlation was found between the hydroxyproline level and patient age. Thus, the content of hydroxyproline was found to be closely related to age and clinicopathologic factors such as the histologic type and degree of lymph node metastasis. PMID- 2552281 TI - [The role of suture material on healing of vascular anastomosis]. AB - The effect of suture material on healing of vascular anastomosis was examined. Four types of vascular grafts, i.e., autogenous vein, preserved human umbilical cord vein, expanded polytetrafluoroethylene and double velour knitted Dacron, were implanted into the abdominal aorta of 78 adult mongrel dogs using two kinds of absorbable sutures (multifilament polyglycolic acid: PGA and monofilament polydioxanone: PDS) and a nonabsorbable suture (polypropylene: PP). The macroscopic findings and the histologic examinations showed that hyalinoid degeneration and calcification resulting from tissue ischemia due to tight and long lasting suture loops interfered with tissue healing at the PP-anastomotic site. On the other hand, fairly good healing of the anastomoses was observed with absorbable sutures because of the reduction of ischemia. The absorbable suture anastomoses could tolerate systemic blood pressure within one month after implantation, and there was no anastomotic disruption at 1000 mmHg pressure in the bursting test 12 months after grafting. From Dec., 1984, 55 anastomoses in 34 bypass-operations employing autogenous vein grafts were performed using PGA and PDS in 28 cases clinically. There were no anastomotic complications. In conclusion, the synthetic absorbable suture material, especially in the form of monofilament, seems to be most suitable for suturing or anastomosing autogenous small vessels at present. PMID- 2552282 TI - [The role of splenectomy in patients with hepatocellular carcinoma and hypersplenism as an aid to hepatectomy]. AB - To evaluate the clinical value of splenectomy for hepatic resection, a total of 20 patients with hepatocellular carcinoma and hypersplenism were examined focusing on a change of total serum bilirubin values after surgery. Both hepatectomy and splenectomy were simultaneously performed in 12 patients, and in 8 patients as a staged operation. Postoperatively, a significant depression of bilirubin values was observed in a group with the preoperative values between 1.0mg/dl and 2.0mg/ml. Three factors (bilirubin, albumin and prothrombin time) in clinical stage were improved just after splenectomy with a statistical significance (p less than 0.05) in a group received staged operation. In 7 out of 8 patients, clinical stages were getting better as one or two stages prior to the hepatectomies. Therefore, we recommend the addition of splenectomy to hepatectomy in the patients whose hyperbilirubinemia are assumed to be correlated with coexisting hypersplenism. PMID- 2552284 TI - [Immunohistochemical studies on laminin in gastric cancer: relationship to histological type and metastasis: preliminary report]. PMID- 2552283 TI - [Effect of sequential MTX/5-FU therapy for a case of disseminated intravascular coagulation syndrome associated with recurrence of gastric cancer--a case report]. AB - Although a 39-year-old male received the curative operation of total gastrectomy for advanced scirrhous carcinoma of the stomach, recurrence of cancer was occurred soon after the surgery, accompanied by hemorrhagic diathesis from DIC. The abdominal CT scan examination revealed the rapid enlargement in the size of the several lymphnodes around the abdominal aorta, and the blood chemistry tests showed marked increase of the serum CEA value. The sequential chemotherapy with intermediate dose of MTX and 5-FU in conjunction with OK-432 was started to treat the case. This chemotherapy was carried out once a week for 5 times and consequently DIC was led to the perfect remission. Furthermore, CEA level decreased within normal range, and the size of the enlarged lymphnodes at paraaortic area diminished remarkably. Although he complained of nausea and loss of appetite during the treatment, no severe adverse effects such as granulocytopenia, diarrhea, or loss of hair were observed. The successful result in this patient suggests that sequential therapy of intermediate dose of MTX and 5-FU with administration of OK-432 may be effective in the treatment of advanced scirrhous carcinoma of the stomach. PMID- 2552285 TI - Synthesis and processing of kinetoplast DNA minicircles in Trypanosoma equiperdum. AB - Kinetoplast DNA, the mitochondrial DNA in trypanosomes, is a giant network containing topologically interlocked minicircles. Replication occurs on free minicircles that have been detached from the network. In this paper, we report studies on the synthesis and processing of the minicircle L and H strands. Analysis of free minicircles from Trypanosoma equiperdum by two-dimensional agarose gel electrophoresis indicated that elongating L strands are present on theta structures. Hybridization studies indicated that L-strand elongation is continuous and unidirectional, starting near nucleotide 805 and proceeding around the entire minicircle. The theta structures segregate into monomeric progeny minicircles, and those with a newly synthesized L strand have a 8-nucleotide gap between nucleotides 805 and 814 (J. M. Ntambi, T. A. Shapiro, K. A. Ryan, and P. T. Englund, J. Biol. Chem. 261:11890-11895, 1986). These molecules are reattached to the network, where repair of the gap takes place. Of the molecules labeled during a 10-min pulse with [3H]thymidine, gap filling occurred on half within about 15 min and on virtually all by 60 min; however, there was no detectable covalent closure of the newly synthesized L strand by 60 min. PMID- 2552286 TI - Identification of two nuclear factor-binding domains on the chicken cardiac actin promoter: implications for regulation of the gene. AB - The cis-acting regions that appear to be involved in negative regulation of the chicken alpha-cardiac actin promoter both in vivo and in vitro have been identified. A nuclear factor(s) binding to the proximal region mapped over the TATA element between nucleotides -50 and -25. In the distal region, binding spanned nucleotides -136 to -112, a region that included a second CArG box (CArG2) 5' to the more familiar CCAAT-box (CArG1) consensus sequence. Nuclear factors binding to these different domains were found in both muscle and nonmuscle preparations but were detectable at considerably lower levels in tissues expressing the alpha-cardiac actin gene. In contrast, concentrations of the beta-actin CCAAT-box binding activity were similar in all extracts tested. The role of these factor-binding domains on the activity of the cardiac actin promoter in vivo and in vitro and the prevalence of the binding factors in nonmuscle extracts are consistent with the idea that these binding domains and their associated factors are involved in the tissue-restricted expression of cardiac actin through both positive and negative regulatory mechanisms. In the absence of negative regulatory factors, these same binding domains act synergistically, via other factors, to activate the cardiac actin promoter during myogenesis. PMID- 2552287 TI - The DAL7 promoter consists of multiple elements that cooperatively mediate regulation of the gene's expression. AB - Expression of the allantoin system genes in Saccharomyces cerevisiae is induced by allophanate or its analog, oxalurate. This work provides evidence for the involvement of distinct types of cis-acting elements in the induction process. The first element was found to have the properties of an upstream activation sequence (UAS). This element was localized to a 16-base-pair (bp) DNA fragment containing a short 5-bp sequence that occurred repeatedly in the upstream region of DAL7. When present in two or more copies, the 16-bp fragment supported high level beta-galactosidase production in a CYC1-lacZ expression vector; there was, however, no response to the allantoin pathway inducer. The second element had the properties of a negatively acting element or upstream repression sequence (URS). This element was localized to a 16-bp DNA fragment containing an 8-bp sequence that was repeated four times in the upstream region of DAL7. A fragment containing the 8-bp repeated sequence placed adjacent to the UAS-containing fragment mediated inhibition of the ability of the UAS to support lacZ expression regardless of whether inducer was present. A third element, designated an upstream induction sequence (UIS), was required for response to inducer. The UIS was localized to a small DNA fragment containing an approximately 10-bp sequence that was repeated twice in the upstream region of DAL7. When a fragment containing the 10-bp repeated sequence was placed adjacent to these UAS and URS elements, the construction (UIS-UAS-URS) supported normal oxalurate-mediated induction of beta-galactosidase synthesis. These data are consistent with the suggestion that multiple, cis-acting elements participate in the induction process. PMID- 2552288 TI - Pancreatic beta-cell-type-specific expression of the rat insulin II gene is controlled by positive and negative cellular transcriptional elements. AB - The insulin gene is expressed almost exclusively in pancreatic beta-cells. The DNA sequences that control cell-specific expression are located upstream of the transcription initiation site. To identify the cis-acting transcriptional control regions within the rat insulin II gene that are responsible for this tissue specific expression pattern, we constructed a series of 5'-flanking deletion mutants and analyzed their expression in vivo in transfected insulin-producing and -nonproducing cell lines. Pancreatic beta-cell-specific expression was shown to be controlled by enhancer sequences lying between nucleotides -342 and -91 relative to the transcription start site. The rat insulin II enhancer appears to be a chimera, composed of a number of distinct cis-acting DNA elements. Both positive and negative transcriptional regulatory elements appear to be responsible for this cell-type-specific expression. We have shown that expression from one element within the enhancer, which is found between nucleotides -100 and -91, is regulated by both positive- and negative-acting cellular transcription factors. Expression from chimeras containing only the enhancer element sequences from -100 to -91 were active only in insulin-producing cells, indicating that the positive-acting factor(s) required for this activity may be active only in beta cells. In contrast to the enhancer region, the rat insulin II gene promoter did not appear to require cell-specific transcription factors. Promoter mutants with 5'-flanking sequences extending to nucleotides -90 and -73 were constitutively active in both insulin-producing and -nonproducing cells. These results suggest that rat insulin II gene transcription in pancreatic beta-cells is imparted by a combination of both negative- and positive-acting cellular factors interacting with the gene enhancer. PMID- 2552289 TI - Depletion of topoisomerase II in isolated nuclei during a glucose-regulated stress response. AB - Conditions, such as anoxia or glucose starvation, which induce the glucose regulated set of stress proteins also lead to resistance to adriamycin (J. Shen, C. Hughes, C. Chao, J. Cai, C. Bartels, T. Gessner, and J. Subjeck, Proc. Natl. Acad. Sci. USA 84:3278-3282, 1987) and etoposide. We report here that chronic anoxia, glucose starvation, 2-deoxyglucose, the calcium ionophore A23187, glucosamine, ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), and tunicamycin (all specific inducers of the glucose regulated system) lead to a rapid and selective depletion of topoisomerase II from isolated nuclei of Chinese hamster ovary cells. This effect precedes a decline in tritiated thymidine incorporation and a redistribution of cells from S into G1/G0. The depletion of the enzyme is not accompanied by a decline in mRNA levels. We have also examined the mutant Chinese hamster K12 cell line which is temperature sensitive for expression of glucose-regulated proteins. When nuclei were isolated from K12 cells incubated at the nonpermissive temperature, a loss of topoisomerase II was again observed in congruence with the expression of stress proteins and cellular resistance to etoposide. These changes were not obtained in parental Wg1A cells incubated at the same temperature. These studies indicate that topoisomerase II is highly sensitive to glucose-regulated stresses and that its depletion from the nucleus, with the associated changes in cell cycle parameters, may represent general characteristics of the glucose-regulated state. Since anoxia and glucose starvation can occur during tumor development, this pathway for expression of drug resistance may have clinical ramifications. PMID- 2552290 TI - Elements involved in S-adenosylmethionine-mediated regulation of the Saccharomyces cerevisiae MET25 gene. AB - In Saccharomyces cerevisiae, the MET25 gene encodes O-acetylhomoserine sulfhydrylase. Synthesis of this enzyme is repressed by the presence of S adenosylmethionine (AdoMet) in the growth medium. We identified cis elements required for MET25 expression by analyzing small deletions in the MET25 promoter region. The results revealed a regulatory region, acting as an upstream activation site, that activated transcription of MET25 in the absence of methionine or AdoMet. We found that, for the most part, repression of MET25 expression was due to a lack of activation at this site, reinforced by an independent repression mechanism. The activation region contained a repeated dyad sequence that is also found in the promoter regions of other unlinked but coordinately regulated genes (MET3, MET2, and SAM2). We show that the presence of the two dyads is necessary for maximal gene expression. Moreover, we demonstrate that in addition to this transcriptional regulation, a posttranscriptional regulation, probably targeted at the 5' region of mRNA, is involved in MET25 expression. PMID- 2552291 TI - Failure to induce a DNA repair gene, RAD54, in Saccharomyces cerevisiae does not affect DNA repair or recombination phenotypes. AB - The Saccharomyces cerevisiae RAD54 gene is transcriptionally regulated by a broad spectrum of DNA-damaging agents. Induction of RAD54 by DNA-damaging agents is under positive control. Sequences responsible for DNA damage induction (the DRS element) lie within a 29-base-pair region from -99 to -70 from the most proximal transcription start site. This inducible promoter element is functionally separable from a poly(dA-dT) region immediately downstream which is required for constitutive expression. Deletions which eliminate induction of RAD54 transcription by DNA damage but do not affect constitutive expression have no effect on growth or survival of noninducible strains relative to wild-type strains in the presence of DNA-damaging agents. The DRS element is also not required for homothallic mating type switching, transcriptional induction of RAD54 during meiosis, meiotic recombination, or spontaneous or X-ray-induced mitotic recombination. We find no phenotype for a lack of induction of RAD54 message via the damage-inducible DRS, which raises significant questions about the physiology of DNA damage induction in S. cerevisiae. PMID- 2552292 TI - Overproduction of yeast viruslike particles by strains deficient in a mitochondrial nuclease. AB - Saccharomyces cerevisiae strains are often host to several types of cytoplasmic double-stranded RNA (dsRNA) genomes, some of which are encapsidated by the L-A dsRNA product, an 86,000-dalton coat protein. Here we present the finding that nuclear recessive mutations in the NUC1 gene, which encodes the major nonspecific nuclease of yeast mitochondria, resulted in at least a 10-fold increase in amounts of the L-A dsRNA and its encoded coat protein. The effect of nuc1 mutations on L-A abundance was completely suppressed in strains that also hosted the killer-toxin-encoding M dsRNA. Both NUC1 and nuc1 strains containing the L-A genome exhibited an increase in coat protein abundance and a concomitant increase in L-A dsRNA when the cells were grown on a nonfermentable carbon source rather than on glucose, an effect independent of the increase in coat protein due to nuc1 mutations or to the absence of M. The increase in L-A expression in nuc1 strains was similar to that observed in strains with mutations in the nuclear gene encoding the most abundant outer mitochondrial membrane protein, porin. nuc1 mutations did not affect the level of porin in the mitochondrial outer membrane. Since the effect of mutations in nuc1 was to alter the copy number of the L-A coat protein genome rather than to change the level of the M toxin genome (as do mak and ski mutations), these mutations define a new class of nuclear genes affecting yeast dsRNA abundance. PMID- 2552293 TI - A 125-base-pair CEN6 DNA fragment is sufficient for complete meiotic and mitotic centromere functions in Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae centromeres contain a conserved region ranging from 111 to 119 base pairs (bp) in length, which is characterized by the three conserved DNA elements CDEI, CDEII, and CDEIII. We isolated a 125-bp CEN6 DNA fragment (named ML CEN6) containing only these conserved elements and assayed it completely separated from its chromosomal context on circular minichromosomes and on a large linear chromosome fragment. The results show that this 125-bp CEN6 DNA fragment is by itself sufficient for complete mitotic and meiotic centromere functions. PMID- 2552294 TI - U5 small nuclear ribonucleoprotein: RNA structure analysis and ATP-dependent interaction with U4/U6. AB - To understand how the U5 small nuclear ribonucleoprotein (snRNP) interacts with other spliceosome components, its structure and binding to the U4/U6 snRNP were analyzed. The interaction of the U5 snRNP with the U4/U6 snRNP was studied by separating the snRNPs in HeLa cell nuclear extracts on glycerol gradients. A complex running at 25S and containing U4, U5, and U6 but not U1 or U2 snRNAs was identified. In contrast to results with native gel electrophoresis to separate snRNPs, this U4/U5/U6 snRNP complex requires ATP to assemble from the individual snRNPs. The structure of the U5 RNA within the U5 snRNP and the U4/5/6 snRNP complexes was then compared. Oligonucleotide-targeted RNase H digestion identified one RNA sequence in the U5 snRNP capable of base pairing to other nucleic acid sequences. Chemical modification experiments identified this sequence as well as two other U5 RNA sequences as accessible to modification within the U5 RNP. One of these regions is a large loop in the U5 RNA secondary structure whose sequence is conserved from Saccharomyces cerevisiae to humans. Interestingly, no differences in modification of free U5 snRNP as compared to U5 in the U4/U5/U6 snRNP complex were observed, suggesting that recognition of specific RNA sequences in the U5 snRNP is not required for U4/U5/U6 snRNP assembly. PMID- 2552296 TI - Reconstitution of the vitamin D-responsive osteocalcin transcription unit in Saccharomyces cerevisiae. AB - The human osteocalcin gene is regulated in mammalian osteoblasts by 1,25(OH)2D3 dependent and -independent mechanisms. The sequences responsible for this activity have been mapped to within the -1339 region of the gene. We show here that this enhancer region functions analogously in Saccharomyces cerevisiae cells engineered to produce active 1,25(OH)2D3 receptor. When fused to the proximal promoter elements of the yeast iso-1-cytochrome c gene, the enhancer demonstrated substantial promoter activity. This activity was elevated further by 1,25(OH)2D3 when the reporter constructs were assayed in cells containing the 1,25(OH)2D3 receptor. This system affords a model for 1,25(OH)2D3 action and represents a simple assay system that will enable definition of the important cis-acting regulatory sequences within the osteocalcin gene and identification of their cognate transcription factors. PMID- 2552295 TI - Isolation and characterization of glucocorticoid- and cyclic AMP-induced genes in T lymphocytes. AB - Glucocorticoids and cyclic AMP exert dramatic effects on the proliferation and viability of murine T lymphocytes through unknown mechanisms. To identify gene products which might be involved in glucocorticoid-induced responses in lymphoid cells, we constructed a lambda cDNA library prepared from murine thymoma WEHI-7TG cells treated for 5 h with glucocorticoids and forskolin. The library was screened with a subtracted cDNA probe enriched for sequences induced by the two drugs, and cDNA clones representing 11 different inducible genes were isolated. The pattern of expression in BALB/c mouse tissues was examined for each cDNA clone. We have identified two clones that hybridized to mRNAs detected exclusively in the thymus. Other clones were identified that demonstrated tissue specific gene expression in heart, brain, brain and thymus, or lymphoid tissue (spleen and thymus). The kinetics of induction by dexamethasone and forskolin were examined for each gene. The majority of the cDNA clones hybridized to mRNAs that were regulated by glucocorticoids and forskolin, two were regulated only by glucocorticoids, and three hybridized to mRNAs that required both drugs for induction. Inhibition of protein synthesis by cycloheximide resulted in the induction of all mRNAs that were inducible by glucocorticoids. Preliminary sequence analysis of four of the 11 cDNAs suggests that two cDNAs represent previously undescribed genes while two others correspond to the mouse VL30 retrovirus-like element and the mouse homolog of chondroitin sulfate proteoglycan core protein. PMID- 2552297 TI - The retinoic acid receptors alpha and beta are expressed in the human promyelocytic leukemia cell line HL-60. AB - The human promyelocytic leukemia cell line HL-60 can be induced to differentiate into granulocytes upon exposure to retinoids. Previously we have shown that extracts of undifferentiated HL-60 cells possess a specific retinoid-binding activity (RSBP-1) corresponding to an approximate 95 kilodalton (kDa) protein as determined by size-exclusion chromatography. We now extend these observations to reveal a second approximate 95 kDa retinoic acid-binding component (RSBP-2), which is separable from RSBP-1 using anion exchange chromatography. We further show that the chromatographic properties of RSBP-1 and RSBP-2 are identical to those found for the retinoid-binding activities present in extracts of HeLa cells transfected with the human retinoic acid receptor (RAR) expression vectors RAR beta phi and RAR-alpha phi, respectively. Moreover, an antiserum preparation directed against RAR-beta selectively immunoprecipitated both the retinoid binding activity in extracts of HeLa cells transfected with RAR-beta phi and that corresponding to RSBP-1 in HL-60 cell extracts. Similarly, an antiserum preparation directed against RAR-alpha immunoprecipitated the retinoid-binding activity in extracts from RAR-alpha phi transfected HeLa cell as well as that corresponding to RSBP-2 in HL-60 cell extracts. Using these antisera, Western blot analyses of extracts from HL-60 cells, and from HeLa cells transfected with either RAR-alpha phi or RAR-beta phi, confirmed that RSBP-2 and RSBP-1 are identical to RAR-alpha and RAR-beta, respectively. However, RAR-alpha, RAR-beta, RSBP-1, and RSBP-2 appeared as an approximate 51 kDa species in sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis in contrast with an apparent approximate 95 k mol wt as estimated from size-exclusion chromatography in the presence of 0.6 M KCl.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552298 TI - Cyclic AMP (cAMP) effects on chorionic gonadotropin gene transcription and mRNA stability: labile proteins mediate basal expression whereas stable proteins mediate cAMP stimulation. AB - Cyclic AMP stimulates a marked accumulation of CG alpha and CG beta mRNAs that reflects, in part, increased rates of gene transcription. We find that a major component of cAMP stimulation of alpha and CG beta mRNAs is independent of new protein synthesis. After treatment of JEG-3 choriocarcinoma cells with cycloheximide, basal levels of alpha and CG beta mRNAs decreased over 12 h to 27% and 13% of control values, respectively. However, cycloheximide treatment did not affect the degree of cAMP-stimulation of alpha and CG beta mRNA levels which increased 20- and 26-fold, respectively. Similarly, cycloheximide did not block cAMP-stimulated transcription of the alpha and CG beta genes. The effect of cAMP treatment on alpha and CG beta mRNA stability was assessed by decay after removal of cAMP, pulse-chase analyses, and decay after inhibition of RNA synthesis by actinomycin D. The half-lives of alpha and CG beta mRNAs determined by decay rates after removal of cAMP were 6.0 h and 7.2 h, respectively. Consistent with these measurements of mRNA stability, alpha and CG beta mRNA half-lives determined by pulse-chase analyses were 8.8 h and 8.6 h, respectively. Cyclic AMP treatment increased the half-lives of alpha and CG beta mRNAs 1.8- and 3.4-fold, respectively. Thus, the effects of cAMP on alpha and CG beta gene expression are predominantly transcriptional, but cAMP also increases mRNA levels via a posttranscriptional mechanism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552299 TI - A novel antibody against a synthetic NH2-terminal myristoyl glycyl src peptide can detect pp60v-src, the transforming protein of Rous sarcoma virus. AB - Novel antibodies were raised against a synthetic NH2-terminal myristoyl(Myr-) tetrapeptide(N-Myr-Gly-Ser-Ser-Lys) which is characteristic of an NH2-terminal portion of pp60v-src, the transforming protein of Rous sarcoma virus. Antisera raised against N-Myr-Gly-Ser-Ser-Lys-haemocyanin reacted with 125I-albumin conjugates with N-Myr-Gly-Ser-Ser-Lys. The immunoreaction was competed for by haemocyanin as well as albumin conjugated with this N-Myr-peptide, while underivatized proteins or an NH2-terminal octapeptide (Gly-Ser-Ser-Lys-Ser-Lys Pro-Lys) had no effect. N-Myr-Gly-Ser-Ser-Lys-(125I)tyramine was also recognized by the antibody. The reaction was competed for by N-Myr-Gly-Ser-Ser-Lys, but not by Gly-Ser-Ser-Lys-Ser-Lys-Pro-Lys. These results suggest a high affinity of the antibody for an N-Myr-peptide moiety. The major (3H)myristate-labelled protein of an apparent molecular weight of 60,000 was detected from chick embryo fibroblasts transformed by Rous sarcoma virus (tsNY68). This protein was demonstrated to possess N-Myr-Gly-Ser-Ser-Lys by the immunoprecipitation and HPLC analyses. Furthermore, the entire circumference of the transformed cells was stained by the antibody upon an immunofluorescent microscopic observation. Thus, these results taken together indicate that the haptenic antibody raised against the myristoyl peptide is useful to detect pp60v-src protein. PMID- 2552300 TI - Rotavirus gene detection with biotinylated single-stranded RNA probes. AB - Biotinylated single-stranded RNA probes from two of the eleven genome segments of the simian rotavirus SA11 were synthesized from cloned DNA and used in dot-blot and Northern-blot hybridization assays. Different types of membranes and conditions to prepare and use synthetic non-radioactive transcript probes were evaluated to obtain optimal test results. Nytran membranes showed the highest sensitivity and lowest backgrounds for hybridization with biotinylated RNA probes. When a gene 6 single-stranded biotinylated probe was used in a dot-blot format, test sensitivity was 0.1 ng for detection of homologous RNA and 0.4-1.5 micrograms for detection of RNA from heterologous rotavirus strains. When used in Northern blots, detection with this gene 6 probe required 1 ng of total SA11RNA or 50 ng of heterologous RNA to be applied to the gels for transfer. Simultaneous hybridization with probes from two different genes on one membrane showed a detection level similar to that seen with single probes alone. The advantages of using biotinylated single-stranded RNA probes to detect or characterize the genes of viruses with double-stranded RNA genomes are shown. PMID- 2552301 TI - Identification of rotavirus genogroups by RNA-RNA hybridization. AB - The genetic relatedness of various human rotavirus strains was examined by RNA RNA hybridization in which 32P-labelled single stranded RNAs produced by in vitro transcription from viral RNAs were used as probes. Denatured genomic double stranded RNAs were hybridized to the probes under highly stringent conditions and the resulting hybrids were fractionated by polyacrylamide gel electrophoresis. Based on the hybridization patterns obtained with probes made from prototype strains Wa (subgroup II, long RNA electropherotype), DS-1 (subgroup I, short RNA electropherotype) and AU-1 (subgroup I, long RNA electropherotype), we have observed that human rotaviruses fall into three distinct gene groups which we have termed 'genogroups'. Identification of genogroups among rotavirus isolates will prove to be a valuable asset for the analysis of naturally occurring reassortants, to trace interspecies transmission of animal rotaviruses to man or vice versa and to identify rotaviruses from environmental sources with regard to their original host species. Furthermore, such an approach will contribute to our understanding of the evolution of rotavirus genes. PMID- 2552302 TI - Use of alkaline northern blot hybridization for the identification of genetic relatedness of the fourth gene of rotaviruses. AB - A recently developed alkaline northern blot hybridization assay (Li, J. K. K., Parker, B. & Kowalik, T., Analytical Biochemistry 163, 210-18, 1987) was used to assess the genetic relatedness of the fourth gene of human rotavirus strains recovered from children with diarrhea and from asymptomatic neonates. Genomic double stranded (ds) RNAs of the rotavirus strains were separated by polyacrylamide gel electrophoresis and were blotted to nylon membranes (Gene Screen Plus or Zeta Probe membranes). The blotted RNAs were then probed with 32P labelled single-stranded (ss) RNA probes prepared by in vitro transcription from single-shelled particles of the different strains. When analysed under a condition of high stringency (52 degrees C, 2.5 x SSC, 50% formamide) that allowed up to 21% of nucleotide mismatch, a high degree of the fourth gene homology was observed among strains recovered from asymptomatic neonates (asymptomatic rotaviruses) or among strains recovered from infants and children with diarrhea (symptomatic rotaviruses), while the homology of the fourth gene between the asymptomatic and symptomatic strains was considerably lower. It is of particular interest that the fourth gene of the AU-1 and AU228 strains recovered from children with diarrhea failed to hybridize to the corresponding gene of either asymptomatic or symptomatic rotavirus strains but showed a high degree of homology with the fourth gene of a feline rotavirus recovered from an apparently healthy cat. These data indicate that a new group of the fourth gene is present among symptomatic rotaviruses and that the fourth gene of this group is genetically related to the corresponding gene of a feline rotavirus. PMID- 2552303 TI - A rapid DNA probe test for detecting human papilloma virus types 6/11 and 16 in biopsy specimens. AB - A simple, rapid and sensitive non-radioactive in situ hybridization assay for human papillomavirus (HPV) has been developed and used to detect HPV 6/11 and HPV16 DNA in ano-genital biopsy specimens. A comparative study to determine the sensitivity and the specificity of this assay relative to Southern blot hybridization assays was performed using one hundred biopsy specimens. The sensitivity of the in situ hybridization assay was 88% for HPV type 6/11 and 89% for HPV type 16. The specificity of the test was 99% for both virus types. In addition to its high sensitivity and specificity, this in situ hybridization assay for HPV uses a non-radioactive detection system. The assay is faster and easier to perform than the Southern blot method. Also, in situ hybridization assays permit the simultaneous evaluation of the histology, as well as the DNA content of biopsy tissues because they do not result in the destruction of tissue or cell morphology. This prototype HPV DNA assay was developed using DNA probes for HPV 6/11 and HPV 16 DNA in anogenital biopsy specimens. However, the protocol developed in these studies can easily be extended to include the use of probes for detecting other HPV types in a variety of tissues. PMID- 2552304 TI - Polyethylene glycol increases specificity of hybridization typing of HPV specimens. AB - Different hybridization conditions for the typing of human papillomaviruses (HPV) in clinical biopsy specimens were tested. The stringency of the hybridization reaction was varied by altering the formamide concentration in the solution. Two polymers, dextran sulphate and polyethylene glycol (PEG), were compared as accelerators of the hybridization reaction. The PEG-containing hybridization solution was found to be suitable for typing clinical HPV specimens. Single stranded RNA probes proved to be more specific than DNA probes in typing clinical specimens. Specimens that were positive both for HPV6 and HPV11 in spot hybridization were confirmed by Southern hybridization. In total, 467 biopsy specimens from genital, anal, oral, aural and nasal lesions were examined for HPV6, HPV11, HPV16 and HPV18 DNA by spot hybridization. Approximately one-third of the specimens were positive for these types. PMID- 2552306 TI - [Ultrastructural analysis of amyloid deposits in pleomorphic adenoma of the salivary glands by topo-optical studies]. AB - It is well established that the stroma of some tumours contains amyloid. Authors have studied in pleomorphic tumours the occurrence of amyloid. Out of 40 salivary gland tumours 5 contained amyloid in their stroma between tumour-cell cords. Amyloid of salivary gland tumours had a resistant structure. The possible pathomechanisms of amyloid deposition are discussed. PMID- 2552305 TI - Production of inositol-phospholipid-derived second messengers in a rat natural killer cell line exposed to susceptible tumor targets. AB - It is currently believed that natural killer (NK) cells kill bound target cells by exocytosis of cytotoxic granules via a calcium-dependent process. After confirming that NK-mediated killing was indeed dependent upon extracellular calcium, we investigated the production of inositol-phospholipid-derived second messengers in a rat NK cell line, RNK, upon exposure to susceptible target cells. These messengers, inositol trisphosphate (IP3) and diacylglycerol (DAG), are associated with calcium-dependent secretory processes in a number of cell types. When RNK cells were exposed to susceptible YAC-1 tumor targets significant amounts of both IP3 and DAG were produced. The levels of the membrane phospholipid parent molecules of these second messengers declined in similarly stimulated RNK cells over a comparable time period. Using three different target cell lines, it was found that the levels of DAG that RNK produced in response to the different targets followed the same rank order as their susceptibility to RNK mediated lysis. These data suggest that IP3 and DAG are produced in NK cells in response to tumor target cells, and these second messengers may have a functional role in NK-mediated killing. PMID- 2552307 TI - Studying DNA mutations in human cells with the use of an integrated HSV thymidine kinase target gene. AB - A shuttle vector carrying the origin of SV40 replication, the thymidine kinase (tk) gene of herpes simplex virus and the E. coli xanthine guanine phosphoribosyl transferase (gpt) gene has been introduced into human TK- cells. A transformed cell line containing only one stably integrated copy of the shuttle vector was used to study mutations in the introduced tk gene at the molecular level. Without selection for gpt expression, spontaneous TK- mutants arose at a frequency of approximately 10(-4)/generation, and were caused by deletion of plasmid sequences. However, when selection for expression of the gpt gene was applied, the background level of mutations at the tk gene was below 4.10(-6). From this cell line, TK- mutants were obtained after treatment with N-ethyl-N-nitrosourea (ENU). COS fusion appeared to be an efficient method for rescue and amplification of the integrated shuttle vector from the human chromosome. After further amplification and analysis in E. coli, rescued tk genes were easily identified and were shown to be physically unaltered by the rescue procedure. In contrast to rescued tk genes from TK+ cells, those obtained from the ENU-induced TK- mutants were unable to complement thymidine kinase-negative E. coli cells. Two such tk mutations were mapped in E. coli by marker rescue analysis. A GC----AT transition was the cause of both mutations. We show here that plasmid rescue by COS fusion is a reliable system for studying gene mutations in human cells, since no sequence changes occurred in rescued DNA except for the 2 ENU-induced sequence changes. PMID- 2552308 TI - The light-dependent cytotoxicity of benzo[a]pyrene: effect on human erythrocytes, Escherichia coli cells, and Haemophilus influenzae transforming DNA. AB - In vitro, the photodynamic compound benzo[a]pyrene (BAP) generates singlet oxygen efficiently when irradiated in organic solvents. It also photogenerates superoxide anion radical in water and can act as a photoreducing agent in the absence of oxygen. In vivo, the hemolysis of human erythrocytes, the inactivation of Escherichia coli cells representing a series of strains differing in excision repair and catalase proficiency, and the inactivation of Haemophilus influenzae transforming DNA activity were used to characterize the phototoxicity of BAP in the presence of near-UV light (290-400 nm). The results are consistent with BAP behaving as a photosensitizer that generates both superoxide and singlet oxygen, and that damages chiefly membranes. DNA does not seem to be a major target in the phototoxic reactions investigated. PMID- 2552309 TI - Induction of cytotoxicity and mutagenesis is facilitated by fatty acid hydroperoxidase activity in Chinese hamster lung fibroblasts (V79 cells). AB - The metabolic activation of benzo[a]pyrene and 7,8-dihydroxy-7,8 dihydrobenzo[a]pyrene was studied in V79 Chinese hamster fibroblasts after supplementations with arachidonic acid or treatments with linoleic acid hydroperoxide. The extent of metabolic activation was estimated using cytotoxicity and mutagenesis as endpoints. Pretreatment of cells with arachidonic acid for 24 h resulted in significant elevations in the content of this fatty acid in cell phospholipids and increased prostaglandin synthesis. Arachidonic acid and linoleic acid hydroperoxide facilitated 7,8-dihydroxy-7,8 dihydrobenzo[a]pyrene cytotoxicity and mutagenesis, and to a lesser extent increased the cytotoxicity and mutagenicity of benzo[a]pyrene. No other compounds tested were mutagenic under these conditions, however, linoleic acid hydroperoxide markedly increased their cytotoxicity. Arachidonic acid-facilitated toxicity and mutagenesis was inhibited by indomethacin, whereas no inhibition was seen when linoleic acid hydroperoxide was used. Nordihyroquairaretic acid abolished the cytotoxicity and mutagenesis facilitated by arachidonic acid and linoleic acid hydroperoxide. Our findings demonstrate that induction of cytotoxicity and mutagenesis following treatment of V79 cells with carcinogens may be limited by low levels of arachidonic acid in these cells. A peroxidatic mechanism is proposed, with limited substrate specificity, for the metabolic activation of chemicals in V79 cells. PMID- 2552310 TI - Evaluation of the mutagenicity and the tumor-promoting activity of parasite extracts: Schistosoma japonicum and Clonorchis sinensis. AB - In relation to the observed association of carcinogenesis with parasitic infections, the mutagenicity of extracts of Schistosoma japonicum and Clonorchis sinensis was examined. In the bacterial mutagenicity tests using the Ames Salmonella typhimurium strains TA98, TA100, TA97 and TA102, and Escherichia coli WP2 and WP2 uvrA pKM101 Schistosoma soluble egg antigen and a homogenate of adult Schistosoma worms showed no positive responses either in the presence or in the absence of S9 mix. Likewise, adult worm extracts of Clonorchis showed no mutagenicity. The Schistosoma soluble egg antigen showed a weak but significant activity for the induction of Epstein-Barr virus expression in viral genome carrying human lymphoblastoid cells in culture. This phenomenon suggests that the soluble egg antigen possesses tumor-promoting activity. PMID- 2552311 TI - Restriction enzyme mapping of ribosomal DNA can distinguish between fasciolid (liver fluke) species. AB - Recognition sites for nine different restriction endonucleases were mapped on rDNA genes of fasciolid species. Southern blots of digested DNA from individual worms were probed sequentially with three different probes derived from rDNA of Schistosoma mansoni and known to span between them the entire rDNA repeat unit in that species. Eighteen recognition sites were mapped for Fasciola hepatica, and seventeen for Fasciola gigantica and Fascioloides magna. Each fasciolid species had no more than two unique recognition sites, the remainder being common to one or both of the other two species. No intraspecific variation in restriction sites was noted in F. hepatica (individuals from 11 samples studied; hosts were sheep, cattle and laboratory animals; geographical origins. Australia, New Zealand, Mexico, U.K., Hungary and Spain), or in F. gigantica (two samples; Indonesia and Malaysia). Only one sample of F. magna was available. One specimen of Fasciola sp. from Japan (specific identity regarded in the literature as uncertain) yielded a restriction map identical to that of F. gigantica. Almost all recognition sites occurred in or near the putative rRNA coding regions. The non transcribed spacer region had few or no cut sites despite the fact that this region is up to about one half of the entire repeat unit in length. Length heterogeneity was noted in the non-transcribed spacer, even within individual worms. PMID- 2552312 TI - Identification and preliminary characterization of cuticular surface proteins of Haemonchus contortus. AB - Cuticular surface antigens of the XL3 and L4 stages of Haemonchus contortus have been studied by surface labeling and immunological techniques. Live worms were labeled with 125I and extracted with sodium dodecyl sulfate (SDS) followed by SDS + 2-mercaptoethanol. The SDS-soluble surface proteins of XL3s and L4s were found to consist of relatively few major species. The pattern of labeled polypeptides was distinctive for each developmental stage. These proteins are refractory to digestion by bacterial collagenase. Several of the proteins are glycosylated. Further extraction of labeled worms with SDS + 2-mercaptoethanol solubilized additional labeled proteins that appeared to be primarily collagens. Rabbit antisera prepared against native XL3 and L4-cuticles reacted strongly with the surfaces of live worms in immunofluorescence assays. In contrast, antisera prepared against SDS-extracted cuticles reacted weakly or not at all with live worms in similar experiments. Rabbit antisera prepared against adult cuticles failed to react with live XL3s or L4s. These studies suggest that the major surface antigens of XL3s and L4s are solubilized by SDS and that there are different antigens present on the cuticular surfaces of XL3s, L4s and adults. Stage-specificity in cuticular surface proteins may contribute to the successful parasitic lifestyle of this nematode. PMID- 2552314 TI - Asbestos-associated diseases in a cohort of cigarette-filter workers. AB - To estimate the effects on health of occupational exposure to crocidolite, a highly toxic form of asbestos, we studied a cohort of 33 men who worked in 1953 in a Massachusetts factory that manufactured cigarette filters containing crocidolite fibers from 1951 to 1957. Twenty-eight of the men have died, as compared with 8.3 deaths expected. This increased mortality was attributable to asbestos-associated diseases. Fifteen deaths were caused by cancer, as compared with 1.8 expected (relative risk, 8.2; 95 percent confidence interval, 4.6 to 13.4), including eight from lung cancer, five from malignant mesothelioma, and two from other types of cancer. There were seven deaths from nonmalignant respiratory disease, as compared with 0.5 expected (relative risk, 14.7; 95 percent confidence interval, 5.9 to 30.3), of which five were due primarily to asbestosis. In contrast, the mortality rates from cardiovascular diseases and all other causes were not increased. Four of the five living workers have pulmonary asbestosis; three of them have recently diagnosed cancers, including two additional lung cancers. We conclude that the extremely high morbidity and mortality in these workers were caused by intense exposure to crocidolite asbestos fibers. PMID- 2552313 TI - Expression of Epstein-Barr virus transformation-associated genes in tissues of patients with EBV lymphoproliferative disease. AB - Epstein-Barr virus (EBV) has been associated with serious or fatal lymphoproliferative disease in immunocompromised patients. EBV nuclear protein 2 and latent membrane protein are characteristically expressed in B lymphocytes proliferating in vitro in response to growth transformation by EBV. These two proteins are thought to be effectors of lymphocyte growth since they increase the expression of B-lymphocyte activation (CD23) and cell-adhesion (LFA 3 and ICAM 1) molecules in vitro. Using monoclonal antibody-immune microscopy, we have demonstrated that these two EBV proteins and their associated B-lymphocyte activation or adhesion molecules are expressed in the infiltrating B lymphocytes in immunocompromised patients with EBV lymphoproliferative disease. These monoclonal antibodies should be useful in the early diagnosis of EBV lymphoproliferative disease and in distinguishing it from other B-lymphocyte cancers associated with EBV, such as Burkitt's lymphoma. The finding of EBV nuclear protein 2 and latent membrane protein and their associated activation or adhesion molecules provides a further pathophysiologic link between EBV and the proliferation of B lymphocytes in immunocompromised patients. PMID- 2552315 TI - Acyclovir to prevent cytomegalovirus infection in renal-graft recipients. PMID- 2552316 TI - Cytomegalovirus and child day care. Evidence for an increased infection rate among day-care workers. AB - To determine whether day-care workers acquire cytomegalovirus infection from the children they care for, we studied 610 women employed at 34 day-care centers over two years. Forty-one percent of the caretakers were seropositive for cytomegalovirus. After adjustment for the effects of race, marital status, and age on seropositivity, the women who cared for children younger than two years of age had a significantly higher seropositivity rate (46 percent) than the women who cared for children older than two years of age (35 percent) (relative risk, 1.29; 95 percent confidence interval, 1.05 to 1.57; P less than 0.02). Of 202 initially seronegative caretakers (observed for an average of 305 days per woman), 19 seroconverted, for an annual seroconversion rate of 11 percent. This rate was significantly higher than the 2 percent annual rate of seroconversion among 229 seronegative women (11 of whom seroconverted) in a comparison group of female hospital employees observed for an average of 781 days per woman (relative risk, 5.0; 95 percent confidence interval, 2.4 to 10.5; P less than 0.001). At three day-care centers in which the children were also studied, seven of the nine women shed isolates of cytomegalovirus in their saliva or urine that had EcoRI and BamHI DNA-digestion patterns identical to the DNA patterns of isolates shed by one or more children in their care. We conclude that workers in day-care centers may acquire cytomegalovirus infection from the children in their care and that this risk is significantly greater among those who care for children less than two years of age. PMID- 2552318 TI - [My patient was crippled and neglected]. PMID- 2552317 TI - Development of a chromosomal DNA probe for the laboratory diagnosis of aspergillosis. AB - Chromosomal DNA was extracted from clinical isolates of Aspergillus fumigatus of human and animal origin using the protoplast lysate method. The probe was developed by the nick translation of the chromosomal DNA genome fraction with p32 as the radiolabel. Hybridization of the probe with endonuclease-cleaved DNA of the same species resulted in a pattern of recognition sites specific for the species. The latter was not seen in other species encountered in clinical specimens. Trials were carried out on sputum experimentally inoculated with the fungus where crude DNA was directly extracted, treated with the endonuclease and hybridized with the probe. The efficacy of the probe was as good with the crude as the purified DNA. The specificity of the probe was determined by testing it against single and mixed DNA populations extracted from different species of several fungal and bacterial genera isolated from and/or known to occur in clinical specimens of respiratory infection origin. The sensitivity of the probe was assessed by detecting a DNA concentration in the specimen equivalent to 3 C.F.U. PMID- 2552319 TI - India: restriction venture. PMID- 2552321 TI - Ubiquitin in a togavirus. PMID- 2552320 TI - Expression of two myogenic regulatory factors myogenin and MyoD1 during mouse embryogenesis. AB - MyoD1 and myogenin are muscle-specific proteins which can convert non-myogenic cells in culture to differentiated muscle fibres, implicating them in myogenic determination. The pattern of expression of MyoD1 and myogenin during the early stages of muscle formation in the mouse embryo in vivo and in limb-bud explants cultured in vitro, indicates that they may have different functions in different types of muscle during development. PMID- 2552323 TI - Moving proteins. PMID- 2552322 TI - Phosphorylation of large tumour antigen by cdc2 stimulates SV40 DNA replication. AB - Simian virus 40 large tumour antigen (T) is a replication origin binding protein required for viral DNA synthesis. Unphosphorylated T antigen is deficient in promoting DNA replication in vitro but can be activated by phosphorylation at residue threonine 124 by the cdc2 protein kinase. This observation demonstrates that T is regulated by phosphorylation and provides a model for cdc2 function in the control of DNA replication. PMID- 2552324 TI - Identification of a novel retinoic acid receptor in regenerative tissues of the newt. AB - In urodele amphibians, the progenitor cells that regenerate amputated limbs (known as the blastema) normally replace only the missing structures. After systemic delivery of retinoic acid (RA), more proximal structures are also formed, indicating that RA can control position specification in the proximal distal axis of the regenerating limb. According to dose and experimental context, retinoids can also re-specify the anteroposterior axis of the limb, induce deletions of skeletal elements, or block re-growth completely. To study the molecular basis of these morphogenetic effects, we screened complementary DNA libraries of newt regenerative tissues (limbs and tails) for hormone nuclear receptors activated by RA. Two functional retinoic acid receptors (RARs) were identified, one of which is the newt homologue of the human alpha-receptor (RAR alpha). The second receptor, called RAR delta, is novel. Sequence analysis suggests that the composite newt RAR previously reported is chimaeric, consisting of 5'RAR-beta-like and 3' RAR delta clones. We conclude that multiple RARs are expressed during limb regeneration in amphibians and suggest that receptor heterogeneity may underlie the different effects of retinoids on limb morphogenesis. PMID- 2552325 TI - RNA editing in wheat mitochondria results in the conservation of protein sequences. AB - RNA editing is a process that results in the production of a messenger RNA with nucleotide sequences that differ from those of the template DNA, and provides another mechanism for modulating gene expression. The phenomenon was initially described in the mitochondria of protozoa. Here we report that RNA editing is also required for the correct expression of plant mitochondrial genes. It has previously been proposed that in plant mitochondria there is a departure from the universal genetic code, with CGG specifying tryptophan instead of arginine. This was because CGG codons are often found in plant mitochondrial genes at positions corresponding to those encoding conserved tryptophans in other organisms. We have now found, however, wheat mitochondrial gene sequences containing C residues that are edited to U residues in the corresponding mRNA sequences. In this way, CGG codons can be changed to UGG codons in the mRNA so that tryptophan may be encoded according to the universal genetic code. Furthermore, for each codon modification resulting from a C----U conversion that we studied, we found a corresponding change in the amino acid that was encoded. RNA editing in wheat mitochondria can thus maintain genetic information at the RNA level and as a result contribute to the conservation of mitochondrial protein sequences among plants. PMID- 2552326 TI - RNA editing in plant mitochondria. AB - A basic principle of molecular biology is that the primary sequence of RNA faithfully reflects the primary sequence of the DNA from which it is transcribed. This concept has been challenged recently by the discovery of RNA editing, broadly defined as any process that changes the nucleotide sequence of an RNA molecule from that of the DNA template encoding it. Examples of RNA editing (see ref. 2 for review) include the insertion and deletion of uridine residues in mitochondrial messenger RNAs in kinetoplastid protozoa, the conversion of a cytidine to uridine in mammalian apolipoprotein-B mRNA, and the appearance of two non-templated guanosine residues in a paramyxovirus transcript. In these cases, RNA editing either re-tailors a non-functional transcript, producing a translatable mRNA, or modifies an already functional mRNA so that it generates a protein of altered amino-acid sequence. Here we report an editing phenomenon that involves the conversion of cytidine to uridine at multiple positions in the mRNA for subunit II of cytochrome c oxidase in wheat mitochondria. Such RNA editing provides an explanation for apparent coding anomalies in plant mitochondria. PMID- 2552327 TI - T cell depletion in transgenic mice carrying a mutant gene for TCR-beta. AB - Classical T lymphocytes recognize foreign antigens in the context of self major histocompatibility complex (MHC) molecules by means of the T-cell receptor (TCR)alpha beta heterodimer. The genes for TCR beta-chains, like immunoglobulin genes, are subject to allelic exclusion. The introduction of a functional TCR beta gene into the germline of mice prevents rearrangement of endogenous TCR-beta genes. Here we report that the introduction of a non-functional TCR-beta genes. Here we report that the introduction of a non-functional TCR-beta gene with a deletion of the major part of the variable region (delta V-TCR-beta), also inhibits endogenous TCR-beta gene rearrangement. This inhibition is mediated via the encoded protein because impairment of endogenous TCR-beta gene rearrangement is not found if a frameshift mutation is introduced into the DJ region of the delta V-TCR-beta transgene. The delta V-TCR-beta transgene can lead to two phenotypes, in which lymphoid development is perturbed. Phenotype A is characterized by a severe impairment of both T and B cell development as reflected by the complete absence of certain lymphoid organs. In phenotype B, lymphoid organs are macroscopically normal, but T cell differentiation is impeded. Virtually all thymocytes lack membrane expression of TCR-alpha beta, but nevertheless carry the CD4 and CD8 antigens (CD4+CD8+ phenotype); they do not, however, mature further. The defect in mice of phenotype B but not of phenotype A can be corrected by the introduction of a functional TCR-beta gene. PMID- 2552328 TI - Lack of effect of opioid peptides, morphine and naloxone on superoxide formation in human neutrophils and HL-60 leukemic cells. AB - There are controversial reports in the literature concerning the effects of opioids on superoxide (O2-) formation in phagocytes, these agents being either inhibitory or stimulatory. We re-examined this issue and compared the effects of the chemotactic peptide, N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMet-Leu Phe), phorbol myristate acetate (PMA), ATP, platelet activating factor (PAF), cytochalasin B (CB) and prostaglandin E1 (PGE1) with those of various opioids on O2- formation in human neutrophils and HL-60 leukemic cells under defined experimental conditions. In the presence of CB, fMet-Leu-Phe and PAF concentration-dependently activated O2- formation in neutrophils with EC50 values of 20 nM and 100 nM, respectively. In the absence of CB, fMet-Leu-Phe and PAF were much less effective. PAF synergistically enhanced O2- formation induced by fMet-Leu-Phe. ATP at a concentration of 100 microM and the opioids, methionine enkephalin, beta-endorphin, dynorphin, [D-Ala2, N-Me-Phe4, Gly5-ol]-enkephalin, [D-Ala2-D-Leu5]-enkephalin and morphine at concentrations between 10 pM to 1 microM did not activate O2- formation. ATP but not beta-endorphin potentiated fMet-Leu-Phe-induced O2- formation. O2- formation induced by a maximally stimulatory concentration of PMA (100 ng/ml) was enhanced by fMet-Leu-Phe but was unaffected by methionine enkephalin or PGE1. PMA at a non-stimulatory concentration (2 ng/ml) potentiated the effect of fMet-Leu-Phe but did not induce responsiveness to PAF, ATP or beta-endorphin. PGE1 strongly inhibited fMet-Leu Phe-induced O2- formation, whereas morphine, methionine enkephalin and the opioid antagonist, naloxone, were without effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552330 TI - How selective is GR 43175? Interactions with functional 5-HT1A, 5-HT1B, 5-HT1C and 5-HT1D receptors. AB - GR 43175 (3-[2-dimethylamino]ethyl-N-methyl-1 H-indole-5 methane sulphonamide) is a novel 5-HT1-like receptor-selective agonist which was reported to be active in the treatment of migraine attacks. The effects of the compound were investigated in radioligand binding studies and in functional models for 5-HT1A, 5-HT1B, and 5 HT1D receptors (inhibition of forskolin-stimulated adenylate cyclase activity in calf hippocampus, rat and calf substantia nigra, respectively) and 5-HT1C receptors (stimulation of inositol phosphate production in pig choroid plexus). GR 43175 displayed the following order of affinity for 5-HT recognition sites (pKD values, -log mol/l, in parentheses): 5-HT1D (7.54) greater than 5-HT1B (6.35) greater than 5-HT1A (6.13) much greater than 5-HT1C (4.13) greater than 5 HT2 (3.67). The same order of potency was observed at functional 5-HT1 receptors, at which GR 43175 acted as a full agonist, with the exception of the 5-HT1C receptor, where the compound was a weak antagonist (pEC50 or pKB values, -log mol/l, in parentheses): 5-HT1D (6.28) greater than 5-HT1B (6.03) greater than 5 HT1A (5.57) much greater than 5-HT1C (4.25). The present data show that GR 43175 interacts preferentially as an agonist with 5-HT1B and 5-HT1D receptors. Since 5 HT1B receptors have not yet been identified in human brain, it seems possible that it is the 5-HT1D receptor which is relevant to the reported antimigraine effects of this compound. PMID- 2552329 TI - LY 83583 (6-anilino-5,8-quinolinedione) blocks nitrovasodilator-induced cyclic GMP increases and inhibition of platelet activation. AB - We studied the effects and the mechanism of action of the cyclic GMP-lowering substance 6-anilino-5,8-quinolinedione (LY 83583) on cyclic GMP-mediated inhibition of platelet function. The activation of washed human platelets by thrombin was counteracted by 8-bromo-cyclic GMP and the direct activators of soluble guanylate cyclase, sodium nitroprusside and endothelium-derived relaxant factor (EDRF = nitric oxide). LY 83583 significantly antagonized the inhibitory effect of sodium nitroprusside and EDRF, but not that of 8-bromo-cyclic GMP, on thrombin-induced aggregation, ATP-release, adhesion to native endothelial cells and increase in concentration of free intracellular calcium ions. In accordance, increases in intracellular cyclic GMP by sodium nitroprusside and EDRF were attenuated by LY 83583. The inhibition of cyclic GMP-mediated effects on platelets by LY 83583 could be related to inhibition of platelet soluble guanylate cyclase, as the activation of the purified enzyme from platelets by sodium nitroprusside was directly inhibited by LY 83583. This effect of LY 83583 was attenuated in the presence of superoxide dismutase. Our findings support the hypothesis that sodium nitroprusside and EDRF inhibit platelet activation by stimulation of soluble guanylate cyclase via nitric oxide. Consequently, inhibition of nitric oxide-induced cyclic GMP formation by LY 83583, which may act by intracellular generation of superoxide anions, facilitates platelet activation. PMID- 2552331 TI - Investigations of the roles of dihydropyridine and omega-conotoxin-sensitive calcium channels in mediating depolarisation-evoked endogenous dopamine release from striatal slices. AB - The relative roles of L- and N-type voltage-sensitive calcium channels (VSCC) in mediating endogenous dopamine release have been investigated by examining the effects of the dihydropyridine (DHP) agonist BAY K 8644 and the antagonist PN 200 110, as well as the VSCC-blocking peptide omega-conotoxin GVIA, on depolarisation evoked dopamine release from superfused rat striatal slices. Dopamine release evoked by electrical field stimulation was virtually unaffected by either of the DHP drugs, but release evoked by raising the K+ concentration to 25 mmol/l was significantly increased by BAY K 8644 and reduced stereospecifically by PN 200 110. Quantitative differences between electrically-evoked and K+-evoked dopamine release with respect to their dependence on extracellular calcium concentration were also observed, with electrically-evoked release requiring higher calcium concentrations. The adenylate cyclase activator forskolin itself increased dopamine release, but did not appear to influence the effectiveness of either DHP drug in altering dopamine release. In contrast to the relatively small effects of the DHP drugs, omega-conotoxin produced a major reduction in electrically-evoked dopamine release as well as a substantial decrease in K+-evoked release. Since omega-conotoxin is thought to block both L- and N-type neuronal VSCC whereas DHP drugs affect only L-type VSCC, these findings suggest that electrically-evoked dopamine release is mediated mainly by calcium influx through N-type VSCC, accounting for the reported lack of effect of many organic calcium antagonists on this process. In contrast, K+-evoked dopamine release appears to involve both L- and N-type VSCC, and can occur at lower extracellular calcium concentrations. PMID- 2552332 TI - [Lepra, you have to keep it in mind!]. PMID- 2552333 TI - [Congenital anomalies of the urogenital tract in patients with a Wilm's tumor]. AB - 133 patients in whom a Wilms' tumour had been diagnosed between 1971 and 1986 were evaluated retrospectively for presence of congenital anomalies of the genitourinary tract. These were detected in 51 patients, 38.3%. A comparison was made between the incidence of each detected malformation in the patient group and in the general population. Some anomalies seemed to have a high association with the nephroblastoma; sharing the same basic embryological disorder is suggested as a possible explanation for this association. PMID- 2552334 TI - [Periampullary carcinoma as a second primary tumor following a previously treated adenocarcinoma of the colon]. AB - Three patients are presented who developed periampullary carcinoma after colectomy for adenocarcinoma. The combination of colonic carcinoma and periampullary carcinoma is rare, although wellknown in patients with colonic polyposis or Gardner's syndrome. Perhaps genetic research might help select patients with a high risk of developing multiple carcinomas in the gastrointestinal tract. The five-year survival rate for periampullary carcinoma after subtotal colectomy is higher than for pancreatic carcinoma. PMID- 2552335 TI - [The role of dietary fiber in gastroenterology: fact or fancy?]. PMID- 2552336 TI - [Amebic dysentery; diarrhea with blood and mucus urges to a study of the feces]. PMID- 2552337 TI - [Is extracranial metastatic malignant glioma iatrogenic?]. AB - A case of an extracranial malignant astrocytoma metastasising to the parotid gland is reported. Cases of extracranial metastasis of a glioma are very rare and largely connected to surgical intervention at the tumour. The impact of surgery on tumour dissemination and the possible pathways of dissemination are discussed. PMID- 2552338 TI - [Physiologic role of the sodium pump. Implications for the study of arterial hypertension]. AB - Mammalian cells have the same hydroelectrolytic composition (high K, low Na), highly different from that of their surrounding. Constancy of cellular composition is insured by the balance between ionic leaks and (Na, K)-pump activity. Ionic leaks, specially sodium, are fundamental. They allow cells to perform a majority of their general and special functions (import of aminoacids, glucose, phosphates; export of acids; nerve influx; muscular contraction; glandular secretion; intestinal and renal reabsorption and secretion). (Na, K) pump is essential to life. It is a kind of general motor that creates and maintains ionic concentrations differences whose potential energy is dissipated by leaks to perform cellular functions. Constancy of hydroelectrolytic intracellular composition hides that leak and pump rates, equivalent between them, are extremely variable among cell types (more than 200 times), and can increase 4 times in less than one minute within a cell type with cellular activity. In a cell, (Na, K)-pump rate is far from maximum velocity. This rate is adjusted nearly instantaneously to balance variations in leak rates; it may undergo short term modulation by endo or exocellular factors; it may undergo long term changes through synthesis of new enzyme molecules. Studies on whole cells of the balance between leaks and pump rates is necessary to understand these cells physiology and pathology. Balance between leaks and (Na, K)-pump activity is altered in renal cells from hypertensive rats, spontaneously (SHR) or genetically selected (Milan Hypertensive Strain: MHS). Strikingly, sodium activation of (Na, K)-pump of inner medullary collecting duct cells of MHS rats is greatly blunted compared to controls. PMID- 2552339 TI - Long-term cortisol treatment impairs behavioral and neuroendocrine responses to 5 HT1 agonists in the rat. AB - The effects of chronic cortisol treatment on neuroendocrine and behavioral responses to serotonin1 (5-HT1) receptor agonists were studied in conscious, freely moving rats. Seven-day cortisol treatment (25 mg/kg/day with osmotic minipumps) markedly suppressed basal plasma corticotropin (ACTH) and corticosterone concentrations, indicating a suppression of the hypothalamo pituitary-adrenocortical axis. Cortisol also decreased body weight, food intake, plasma norepinephrine (NE), and epinephrine (E) levels. In the drug challenge studies, we used two 5-HT1 agonists, the 5-HT1B and 5-HT1C agonist, m chlorophenylpiperazine (m-CPP), and the 5-HT1A agonist, 8-hydroxy-2-(di-n propylamino) tetralin (8-OHDPAT), to examine the effect of cortisol on their behavioral and neuroendocrine effects. After 7-day cortisol treatment, plasma prolactin responses to both m-CPP and 8-OHDPAT were significantly decreased. While the plasma NE, E, and food intake responses to m-CPP were also significantly reduced by cortisol treatment, these same responses to 8-OHDPAT were unchanged. The effect of m-CPP on locomotor activity was also decreased. Since only the responses to m-CPP and 8-OHDPAT previously shown to be antagonized by pretreatment with the 5-HT1/5-HT2 antagonist, metergoline, were significantly attenuated after cortisol treatment, these changes may be specific to 5-HT receptors. These data indicate that chronic exposure to high glucocorticoid levels alters 5-HT1 receptor-mediated functions and provides additional evidence relevant to the contribution of glucocorticoid elevation to the symptoms of depression. PMID- 2552340 TI - Modulation of the neonatal pituitary and adrenocortical responses to stress by thyroid hormones in the rat: effects of hypothyroidism and hyperthyroidism. AB - Neonatal rats exhibit a period of diminished pituitary and adrenocortical responses to stress during the first 2 weeks of life. Since thyroid hormones are known to affect brain development, modulation of these responses to stress by alterations in thyroid hormone status have been investigated in hypothyroid (Hypo) and hyperthyroid (Hyper) rat pups. Changes in ACTH and corticosterone (B) levels were measured under basal and stress conditions (3 min exposure to ether vapors) in neonates of various ages (day 5-21). Basal T4 and corticosterone binding globulin (CBG) levels were also measured. Hypo pups were obtained from methimazole-treated mothers and hyperthyroidism was induced by daily subcutaneous injections of L-T4 (100 micrograms/kg BW) from birth on. In Hyper rats, premature onset of ACTH and B responses to stress was observed in 5-day-old rats while significant ACTH and B secretion only appeared by day 10 in vehicle-injected rats. By contrast, ACTH and B responses to stress were delayed in Hypo pups and only occurred by day 21. The lack of ACTH and B responses to stress of 14-day-old Hypo rats could be reversed by one single L-T4 injection (100 micrograms/kg BW) given 24 h, but not 4 h prior to exposure to stress. On day 21, smaller (p less than 0.05) stress-induced ACTH release was observed both in Hypo and Hyper rats compared to intact rats, concomitant with a diminished ACTH secretion following exogenous ovine CRF (10 micrograms/kg BW, i.p.) administration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552341 TI - Seasonal modification of ovine pineal function. 2. Steroidal effects on melatonin and prolactin profiles. AB - Pineal beta-adrenoceptor density and affinity in ewes are modified in a season dependent manner by gonadal steroids and by the sympathetic innervation of the gland. The present study was undertaken to relate the steroidal effects on the receptors to post receptor endocrine events, and to investigate the influence of the sympathetic innervation of the pineal gland on these events. Plasma melatonin and prolactin profiles were determined during anestrus and during the normal breeding season in ewes subjected to sympathetic denervation of the pineal and/or a range of steroid-related treatments. Wherever valid comparisons could be drawn between effects of the treatments on beta-adrenoceptor variables and on circulating hormone levels, similar effects were noted. Further, ganglionectomy influenced hormone profiles similarly to estradiol under all conditions tested. It appears that gonadal steroids (estradiol) and the sympathetic neurotransmitter noradrenaline have opposing actions on prolactin levels, just as they have on pineal beta-adrenoceptor binding affinity. These findings suggest that steroid mediated changes in receptor number and affinity are reflected in post receptor endocrine events. In addition, other factors (e.g. photoperiodic information transmitted via the sympathetic innervation) also play important roles in the regulation of the observed hormonal profiles. A steroid-mediated feedback regulation of pituitary prolactin release, partly direct and partly via pineal melatonin release, is suggested. PMID- 2552342 TI - Yohimbine increases cerebrospinal fluid and plasma norepinephrine but not arginine vasopressin in humans. AB - Previous studies have demonstrated alpha 2-inhibitory regulation of central nervous system (CNS) noradrenergic and arginine vasopressinergic systems. We tested the hypothesis that alpha 2-inhibition of CNS noradrenergic and vasopressinergic systems is tonic in nature by measuring the response of cerebrospinal fluid (CSF) norepinephrine (NE) and arginine vasopressin (AVP) to the alpha 2-antagonist yohimbine in 7 young normal male human subjects. We also evaluated the tonic nature of alpha 2-inhibition of the sympathetic nervous system (SNS) and of AVP release into plasma by measuring the response of plasma NE and plasma AVP to yohimbine. CSF NE was significantly higher following yohimbine as compared to placebo. In contrast CSF AVP did not differ between yohimbine and placebo conditions. Similarly, plasma NE was significantly higher following yohimbine as compared to placebo, while plasma AVP was unchanged. These results support a tonic alpha 2-inhibitory regulatory mechanism for both CNS noradrenergic systems and sympathetic outflow. Such tonic alpha 2-inhibition could not be demonstrated for regulation of AVP levels in CSF or plasma in humans. PMID- 2552343 TI - Regulation of vasopressin receptors and phosphoinositide hydrolysis in the septum of heterozygous and homozygous Brattleboro rats. AB - Accurel polypropylene mini-devices, loaded with arginine vasopressin (AVP) and implanted in the lateral cerebral ventricle were used to centrally treat heterozygous (HE) and homozygous (HO) Brattleboro (BB) rats. After 1 week of treatment, the concentration of AVP receptors in the HO-BB rat septum decreased from 19.4 +/- 2.6 to 12.4 +/- 1.1 fmol/mg protein, but remained unchanged in the HE-BB rat (10.7 +/- 0.8 and 7.0 +/- 1.1 fmol/mg protein). In the HO-BB rat the [3H]-AVP equilibrium dissociation constant (KD) of the septal AVP receptor decreased following AVP treatment (from 4.17 +/- 0.7 to 1.97 +/- 0.3 nM) compared to that of control animals. This decrease in receptor number following AVP treatment was accompanied by a decrease in the postreceptor response to AVP as measured by the AVP-stimulation of [3H]-inositol-1-phosphate (IP1) accumulation (22.0 +/- 6.1%) when compared to untreated animals (54.3 +/- 8.3%). This apparent AVP-induced down-regulation was not due to occupancy of the binding sites by AVP since preincubation of the tissue at 37 degrees C for 60 min (which was able to cause near-complete dissociation of the hormone-receptor complex) did not result in an increased number of binding sites upon reexposure to [3H]-AVP. This study thus provides evidence for the homologous down-regulation and desensitization in terms of [3H]-IP1 accumulation (phosphoinositide hydrolysis) of AVP receptors in the septum of the BB rat. PMID- 2552344 TI - Regulation of high-affinity GABAa receptors in specific brain regions by ovarian hormones. AB - The regulation of 3H-muscimol binding to high-affinity GABAa receptors by estradiol (E) and by progesterone (P) was studied within discrete brain regions using in vitro quantitative autoradiography. Treatment of ovariectomized and adrenalectomized female rats with E resulted in a decrease of muscimol binding only in specific estrogen-sensitive brain regions like the ventromedial nuclei (VMN) of the hypothalamus, the arcuate nucleus (ARC), the medial amygdala and the midbrain central grey (MCG). When administered alone, P had no effect. However, in estrogen-primed females, P increased muscimol binding in both VMN and MCG to levels seen in control animals. Thus, E and P exert opposite effects on the GABAa receptor within these two nuclei. As both hormones facilitate female reproductive behavior as well as the release of luteinizing hormone, present results suggest that E and P affect muscimol binding by different mechanisms. PMID- 2552345 TI - Guanine nucleotides regulate the affinity of melatonin receptors on the ovine pars tuberalis. AB - The effect of guanine nucleotides and related analogues on the binding of 2 [125I]-melatonin to membranes prepared from ovine pars tuberalis was studied. Dose-dependent inhibition of 2-[125I]-melatonin binding was observed, with an order of potency of GTP gamma S much greater than Gpp(NH)p greater than GTP = GDP. GMP, cyclic GMP and ATP had negligible effects. Analysis of saturable binding revealed that GTP gamma S (1 microM) promoted an apparent reduction in receptor density of about 50%, without a concomitant change in receptor affinity. These results are consistent with a melatonin receptor existing in an equilibrium between high- and low-affinity states, with GTP and related analogues able to cause a shift in the equilibrium in favour of the lower-affinity form. The sensitivity of 2-[125I]-melatonin binding to guanine nucleotides implies the presence of a melatonin receptor on the ovine pars tuberalis, the action of which is mediated via a G protein. PMID- 2552346 TI - Myelography in patients with acquired immuno deficiency syndrome. Indications and results. AB - Neurological complications in patients with Acquired Immuno Deficiency Syndrome (AIDS) are frequent and in addition to central nervous system syndromes, involvement of the peripheral nervous system is increasingly seen. We evaluated the indications and results of myelographic examination in six AIDS-patients with signs of peripheral nervous system disease, out of 200 AIDS-patients with neurological complications. Five of these patients had a polyradiculopathy, with proven cytomegalovirus (CMV) infection in four cases. There were two abnormal myelographic examinations with findings of cauda equina nerve root involvement, both in patients with proven CMV-polyradiculopathy. These abnormal findings had no direct therapeutic consequences. Myelography is not essential for establishing the diagnosis, which is based on cerebrospinal fluid (CSF) analysis, but may be indicated to exclude a spinal cord or nerve root compressive lesion. PMID- 2552347 TI - Regulation of endogenous acetylcholine release from mammalian brain slices by opiate receptors: hippocampus, striatum and cerebral cortex of guinea-pig and rat. AB - The effects of opiate agonists on acetylcholine release from hippocampal, striatal and cerebral cortical slices were tested; tissue from rat was compared to that from guinea-pig. The results show that opiate receptors in each of these areas can alter the evoked release of acetylcholine from nerve terminals; however, there are species and tissue differences with respect to the apparent subtype of opiate receptor effective. In the hippocampus and striatum of the two species studied, opiates caused a dose-dependent decrease in evoked acetylcholine release from tissue slices but in the guinea-pig kappa-selective agonists were effective, and mu or delta agonists were not, whereas in the rat, mu-, but not delta- or kappa-selective drugs were effective. Opiates also altered acetylcholine release from the frontal, parietal and occipital cortex of both of these species. In all three regions of the guinea-pig cortex, kappa and delta agonists were active and in the parietal cortex mu agonists were also active; rat cortical slices showed similar results except that delta agonists were not effective. The inhibitory effects of the opiate agonists were effectively antagonized by the non-selective opiate antagonist naloxone and by the calcium channel agonists, BAY K 8644 or YC-170. In addition, the effects of the opiate drugs tested in this study on acetylcholine release were confined to evoked release, that is, spontaneous acetylcholine release was not affected. The results suggest that in guinea-pig and rat brain, opiate receptors regulate acetylcholine release, and that, although the subtypes of opiate receptors involved in this effect are different in the two species and in different tissues from the same species, the effect results from a common mechanism that involves alterations of calcium influx into the nerve terminals during depolarization. PMID- 2552348 TI - A possible pacemaker mechanism in pars compacta neurons of the guinea-pig substantia nigra revealed by various ion channel blocking agents. AB - The membrane properties of pars compacta neurons in the in vitro guinea-pig substantia nigra have been studied in the presence of sodium, calcium and potassium channel blockers. The following properties, which have already been described for dopamine-containing substantia nigra zona compacta neurons were observed: high and low threshold calcium spikes; a calcium-activated potassium mediated transient; inward rectification. Inward rectification was sensitive to caesium ions. An additional property was seen reminiscent of an "A" current, although resistant to 4-aminopyridine. It is suggested that this outward transient is in fact a calcium activated potassium conductance. Under certain conditions calcium-mediated rhythmic depolarizations were observed. It is suggested that at least two of the properties seen (outward rectification and low threshold calcium spike) could interact to provide the basis for a pacemaker mechanism in pars compacta neurons. PMID- 2552349 TI - Autoradiographic localization of binding sites for vasoactive intestinal peptide and angiotensin II on neurons and astrocytes of cultured rat central nervous system. AB - The cellular localization of binding sites for [125I] vasoactive intestinal polypeptide and [3H]angiotensin II was studied in explant cultures of rat spinal cord, brain stem, cerebellum and cortex by means of autoradiography. In spinal cord cultures, interneurons of the dorsal horn and motoneurons of the ventral horn were labelled by [125I]vasoactive intestinal polypeptide and [3H]angiotensin II. In many brain stem cultures, groups of large neurons revealed intense binding of both peptides. In contrast, only few medium-sized cerebellar neurons, probably interneurons, showed binding sites for [125I]vasoactive intestinal polypeptide and [3H]angiotensin II. Furthermore, the intensity of labelling of cerebellar neurons was usually weaker than that of neurons of the brain stem and spinal cord. Many neurons in cultures of neocortex were also labelled by [125I]vasoactive intestinal polypeptide, whereas little binding was found with [3H]angiotensin II. In addition to neurons, binding sites for both peptides were also observed on astrocytes. Labelling of these cells was more intense in spinal cord and brain stem cultures than in cultures of cerebellum and cortex, suggesting that only a certain type or a certain population of astrocytes possesses receptors for vasoactive intestinal polypeptide and angiotensin II, or that glial cells in different parts of the CNS have different physiological and pharmacological properties. PMID- 2552350 TI - Central-type benzodiazepines and the octadecaneuropeptide modulate the effects of GABA on the release of alpha-melanocyte-stimulating hormone from frog neurointermediate lobe in vitro. AB - The involvement of the GABA-benzodiazepine receptor complex in the regulation of melanotropin secretion has been investigated using perfused frog neurointermediate lobes. The GABAA agonist 3-amino-1 propane sulfonic acid mimicked the biphasic effect of GABA on alpha-melanocyte-stimulating hormone secretion: a brief stimulation followed by an inhibition of melanotropin secretion. The GABAA antagonist SR 95531 (10(-4) M) inhibited both stimulation and inhibition of alpha-melanocyte-stimulating hormone release induced by GABA (10(-4) M). Since the inhibitory effect of baclofen (10(-4) M) was partially antagonized by SR 95531 (10(-4) M), it appears that the GABAergic control of alpha-melanocyte-stimulating hormone release is mainly achieved through activation of GABAA receptors. GABA-induced stimulation of alpha-melanocyte stimulating hormone release was inhibited by tetrodotoxin (10(-5) M), an Na+ channel blocker, or nifedipine (10(-5) M), a voltage-dependent Ca2+ -channel blocker, suggesting that Na+ and Ca2+ ions are involved in the stimulatory phase of GABA action. Only central-type benzodiazepine binding site agonists such as clonazepam (10(-4) M) modified alpha-melanocyte-stimulating hormone release. In fact, clonazepam (10(-7) to 10(-5) M) led to a dose-dependent potentiation of both GABA-induced stimulation and inhibition of alpha-melanocyte-stimulating hormone release. This potentiating effect was antagonized by the GABAA antagonist SR 95531 (10(-4) M) or by the central-type benzodiazepine binding site antagonist flumazenil (10(-4) M), whereas picrotoxin (10(-4) M) abolished only the stimulatory phase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552352 TI - Lumbosacral plexopathy secondary to perirectal abscess in a patient with HIV infection. PMID- 2552351 TI - The dorsal columns. PMID- 2552353 TI - The biological fate of 14C-dimercaptosuccinic acid in monkeys and rabbits. AB - The biological fate of 14C-labeled dimercaptosuccinic acid (DMSA) in monkeys and rabbits was determined by measuring the 14C activity in their urine, feces, and expired air (14CO2). Monkeys absorbed less than 20% DMSA from three oral dose levels (0.082, 0.16, and 0.5 mmol/kg) of 14C-DMSA, and the rabbits absorbed 32% DMSA or less from an oral dose of 14C-DMSA (0.5 mmol/kg). Although the bioavailability of DMSA was limited in either species, DMSA was detected in the blood of both species within minutes after oral dosing. In either species, most of the radiolabel from the absorbed 14C-DMSA was detected in the urine within 12 hours. We also developed a sensitive assay for directly measuring levels of DMSA (as free thiols) in blood. Intact DMSA was not detected in the blood of the monkeys or the rabbits more than 200 minutes after oral or intravenous dosing at 0.5 mmol DMSA/kg body weight. However, 14C activity in blood and urine of the monkeys was measurable 72 hours after this dose. Differences between measured 14C concentrations and intact DMSA concentrations in the blood suggest the presence of DMSA metabolites that have longer half-lives than DMSA. Consequently, until the biological activities of these compounds are identified, the pharmacokinetic analysis of DMSA may be incomplete. PMID- 2552354 TI - Hepatitis A: a serological study in the Canadian Armed Forces. AB - Sera were collected from 3958 Canadian Forces personnel, including recruits, personnel posted to bases in Canada and on rotation with United Nations Peacekeeping forces, and the crew of a Canadian destroyer. All sera were tested for the presence of anti-hepatitis A antibodies by a competitive radioimmunoassay. The overall prevalence of anti-hepatitis A antibodies was found to be 23.5% (3.2% to 40.0%). The incidence of viral hepatitis A, as measured by seroconversion in paired sera, was found to be 0.17% (0.01% to 1.42%). The study results support the existence of a direct relationship between increasing age and an increased prevalence of anti-hepatitis A antibodies. PMID- 2552355 TI - [Psammomatous somatostatinoma of Vater's papilla. A case report]. PMID- 2552356 TI - [The multicentricity of minimal breast cancer. The observations and therapeutic implications apropos 30 cases]. PMID- 2552358 TI - [Phyllodes tumors of the breast. Clinico-statistical data from the division of Obstetrics and Gynecology of the S. Giacomo Hospital for the years 1981-1985]. AB - In the light of twenty cases encountered in only five years, the peculiar characteristics of cystosarcoma phyllodes of the breast are described, with an analysis of its morphological, diagnostic, therapeutic and prognostic aspects. PMID- 2552357 TI - [Phyllodes tumor of the breast]. PMID- 2552359 TI - [Role of viral infections in the genesis of dysplasias of the uterine cervix]. AB - A study of 80 cases of dysplasia of the uterine cervix showed the coexistence of HPV infections in 52 cases. This association (65%) shows the important role played by the human papilloma virus in the genesis of cancer of the cervix. PMID- 2552360 TI - FDA expands testing of anti-AIDS drugs. PMID- 2552361 TI - Oral hairy leukoplakia-like lesions in an HIV-negative male: a case report. AB - Oral hairy leukoplakia is generally found only in HIV-seropositive individuals or in those who are at risk of developing this infection; its occurrence is accepted to be a reliable predictor for the development of AIDS in those patients who are not already suffering from this disease. This report describes an 18-year-old male who was proven to be HIV-seronegative, was not in any of the at-risk groups associated with this infection, but who had the typical features of oral hairy leukoplakia, clinically and histologically. In such individuals these lesions should be described as hairy leukoplakia-like lesions. PMID- 2552362 TI - Intracellular localization of 99Tcm-d,l-HMPAO and 201Tl-DDC in rat brain. AB - The intracellular localization and relative distribution of 99Tcm hexamethylpropyleneamine-oxime (99Tcm-d, l-HMPAO) and 201Tl diethyldithiocarbamate (201Tl-DDC), which have been used to assess regional cerebral blood flow (rCBF) in man, were investigated in rat brain. 99Tcm-d, l HMPAO was found attached mainly to cell organelles whilst 201Tl-DDC was localized mainly in free state in the cytosolic fraction. Isolation of neuronal and glial nuclei showed that there is higher uptake of 99Tcm-d, l-HMPAO in neuronal nuclei than in glial nuclei for all the different regions of the rat brain investigated. These data demonstrate a different subcellular localization of the two lipophilic agents studied. In view of their proposed clinical utility, this difference merits further investigation. PMID- 2552363 TI - Technetium-labelled bran to measure solid gastric emptying. PMID- 2552364 TI - 99Tcm-labelled meso-HMPAO and glutathione content of human lung tumours. AB - Levels of reduced glutathione (GSH) are increased in some types of malignant tumours and are known to influence the response to radio- and chemotherapy. In vitro studies suggest a correlation between cellular GSH concentration and retention of the meso form of hexamethyl propyleneamineoxime (HMPAO). This study investigates the relationship between in vivo tissue retention of 99Tcm-labelled HMPAO and GSH concentration in ten patients referred for thoracotomy for possible lung cancer. Retention of 99Tcm-HMPAO was measured using single photon emission computed tomography (SPECT). The tumour and normal lung concentration of reduced glutathione (GSH) was measured in tissue specimens collected peroperatively. Malignancy was confirmed in eight patients. Of seven patients undergoing curative resection for carcinoma, tumour GSH concentration was higher (mean 2.76 mM) than normal lung (mean 1.04 mM). In one neurofibroma, the GSH concentration was 1.80 mM. No correlation was found between 99Tcm meso-HMPAO retention and either the tumour GSH concentration or the tumour:lung GSH ratio. The results from this small series demonstrate that the intracellular GSH concentration of malignant lung tumours is generally higher than that in normal lung but that meso-HMPAO retention could not be used to predict these levels. PMID- 2552365 TI - Interrelationship between insulin, dietary fiber, and exercise in the management of pregnant diabetics. AB - The metabolic changes during pregnancy are characterized as "accelerated starvation" and "facilitated anabolism" which are, in turn, worsened by the presence of diabetes mellitus. The current roles of dietary fiber and exercise as adjunctive therapeutic modalities in the management of diabetic pregnancies are discussed herein. PMID- 2552366 TI - Bacteria and inflammatory cells reduce chorioamniotic membrane integrity and tensile strength. AB - Bacteria and the inflammatory response they engender are implicated in the pathophysiology of premature rupture of fetal membranes and preterm birth. We tested the hypothesis that bacteria and polymorphonuclear neutrophils may have both separate and combined effects in weakening the amniochorionic membrane, and thus predispose to premature rupture of membranes. We examined how three parameters of membrane integrity (bursting tension, work to rupture, and elasticity) were affected by standardized preparations (10(9) cfu/mL) of group B streptococci or Staphylococcus aureus in the presence or absence of purified human neutrophils (32 x 10(6)/mL). In addition, effects of purified human neutrophil elastase were evaluated. Exposure to either group B streptococcus or S aureus decreased membrane strength, elasticity, and work to rupture. Only activated neutrophils had significant effects on membrane strength. Membrane exposure to S aureus plus neutrophils led to an additive weakening of fetal membranes. On the other hand, group B streptococci did not interact with neutrophils to yield a significant further weakening of the membranes. Elastase (150 U/mL) also weakened the membrane. The results were correlated with measures of protease (Azocoll and ninhydrin assays) and peroxidase (dimethoxybenzidine) activity. Our findings support the concept that human neutrophils and their constituent enzymes may act in concert with bacteria and their protease(s) in weakening amniochorion, and may possibly predispose to premature rupture of membranes in some women. These observations require clinical correlations. PMID- 2552367 TI - A comparison of slot blot, southern blot, and in situ hybridization analyses for human papillomavirus DNA in genital tract lesions. AB - Genital tract lesions were analyzed for human papillomavirus (HPV) DNA by in situ hybridization using probes of HPVs 6/11, 16/18, and 31/33/35. All of the HPVs detected in vulvar and perianal condylomata by in situ hybridization were HPV 6/11-related, whereas the majority of HPVs detected in cervical intraepithelial grade I lesions were types 16/18- and 31/33/35-related. None of the lesions with histologic features equivocal for HPV infection had detectable HPV DNA by in situ hybridization, though some did contain HPV DNA sequences as ascertained by filter hybridization analysis. The sensitivity of in situ hybridization was compared with that of filter hybridization (slot blot and/or Southern blot). The correlation was high (28 of 30) for cases that contained HPVs 6/11 or 16, as deduced by filter hybridization, and was much less (ten of 29) for cases that contained HPVs distinct from types in the filter hybridization probe cocktail (HPVs 6/11, 16, 18, 31, 35 and 51). There was a high concordance between the results of Southern blot hybridization and slot blot hybridization analyses, especially with cases that contained HPVs 6/11 and 16. In situ hybridization, slot blot, and Southern blot hybridization analyses are all very effective in detecting the common HPV types (HPVs 6/11 or 16). In situ hybridization is useful in differentiating cervical lesions that contain HPV 6/11 from those that contain HPV 16 or other types with oncogenic potential. However, filter hybridization is superior to in situ hybridization when analyzing cases with histologic findings equivocal for HPV infection or cases that contain HPV types related to, but distinct from, the types included in the probe. PMID- 2552368 TI - Poland-Mobius syndrome: a case report. AB - The Poland-Mobius syndrome is a combination of two rare congenital syndromes with an estimated prevalence of 1:500,000. It is characterized by a nonprogressive bilateral facial paralysis, the inability of the eyes to abduct beyond the midline, orofacial anomalies, limb deficiencies, and an absence or hypoplasia of the pectoral muscles. PMID- 2552369 TI - Herpes simplex virus enhances the 7,12-dimethylbenz[a]anthracene (DMBA)-induced carcinogenesis and amplification and overexpression of c-erb-B-1 proto-oncogene in hamster buccal pouch epithelium. AB - We studied the effects of herpes simplex virus type 1 (HSV-1) inoculation and topical 7,12-dimethylbenz[a]anthracene (DMBA) application, alone or in combination, on the carcinogenesis and on the amplification and expression of various cellular proto-oncogenes in hamster buccal pouch tissue. Topical DMBA treatment produced tumor formation in pouches, but HSV-1 inoculation, alone caused no neoplastic changes. In pouch tissues receiving both DMBA application and HSV-1 inoculation, the development of initial leukoplakia and tumor has hastened and enhanced in comparison with those receiving DMBA alone. Topical DMBA application to pouch tissue induced an amplification and an increase in the expression of cellular erb-B-1 (c-erb-B-1) proto-oncogene in the epithelial tissue, whereas repeated infection with HSV-1 alone did not. Topical DMBA combined with HSV-1 inoculation, however, resulted in greater amplification and expression of c-erb-B-1 proto-oncogene in the pouch epithelial tissue compared to the DMBA alone. These data indicate that HSV-1 inoculation significantly increases the carcinogenic activity of DMBA, in part, by probably enhancing DMBA induced amplification and expression of c-erb-B-1 proto-oncogene in hamster buccal pouch tissue. PMID- 2552370 TI - Malignant fibrous histiocytoma of the mandible. AB - Malignant fibrous histiocytoma of the mandible has appeared frequently enough in the world literature in recent years to assume a legitimate place in the differential diagnosis of neoplastic masses of the lower jaw. This article reports a pertinent case and tabulates and correlates the findings of all cases reported thus far. The report also explores the contribution of immunohistochemistry to proper diagnosis and emphasizes the advantages of a cojoint effort between surgeon and pathologist at the time of initial patient evaluation. The case reported also demonstrates the poor prognostic characteristics of this lesion and the uncertainty as to proper mode of treatment. PMID- 2552371 TI - Benign fibrous histiocytoma of the maxilla. AB - Benign fibrous histiocytomas of bone are unusual neoplasms that often are confused with metaphyseal fibrous defects. Although the two lesions have overlapping microscopic characteristics, they differ in their clinicopathologic presentations. This report describes the clinicopathologic features of the first reported case of benign fibrous histiocytoma involving the maxilla and the fourth description of this tumor in the jawbones. In addition, the concept of fibrohistiocytic lesions of bone is discussed with consideration of their pathologic classification and their clinical, radiographic, and microscopic differential diagnosis. PMID- 2552372 TI - Strategies to reduce hospital readmissions: a review. PMID- 2552374 TI - The c-erbA alpha-encoded thyroid hormone receptor is phosphorylated in its amino terminal domain by casein kinase II. AB - The c-erbA alpha progenitor of the v-erbA oncogene of avian erythroblastosis virus (AEV) encodes a nuclear receptor for the thyroid hormone triiodothyronine (T3) which acts as a ligand-dependent transcription factor. As previously reported (Goldberg et al., EMBO J., 7, 2425-2433), the 46 kd chicken c-erbA alpha encoded T3 receptor (ck-ErbA alpha) is phosphorylated at two major sites. Only one of these sites (Ser28/Ser29) is retained in the v-erbA-encoded P75gag-v-erbA protein. We report here the identification of the second phosphorylation site of ck-ErbA alpha as a single serine residue localized at position 12. We propose that casein kinase II, a protein kinase distributed in the cytosolic and nuclear compartments of a number of different tissues, is responsible for serine 12 phosphorylation on the following grounds. First, serine 12 is part of a sequence containing multiple acidic amino-acids, a feature common to all sites phosphorylated by casein kinase II in physiological substrates. Second, ck-ErbA alpha was found to be phosphorylated by purified casein kinase II in vitro at the same site, as defined by two-dimensional mapping experiments, as that observed in vivo. Third, conversion of serine 12 into an unphosphorylatable alanine residue by site directed mutagenesis abolishes the phosphorylation of ck-ErbA alpha by casein kinase II in vitro. Phosphorylation of serine 12 is likely to play a role in the modulation of ErbA alpha function since both serine 12 and the casein kinase II phosphorylation sequence motif are phylogenetically conserved in all known members of the c-erbA alpha gene family encoding T3 binding proteins. The codon specifying serine 12 in ck-ErbA alpha being precisely the point where recombination between gag and ck-c-erbA alpha occurred to generate v-erbA, our results furthermore suggest that deletion of serine 12 could contribute to the oncogenic activation of v-erbA. PMID- 2552373 TI - Nerve growth factor-induced differentiation in PC-12 cells is blocked by fos oncogene. AB - Rat pheochromocytoma (PC-12) cells respond to nerve growth factor (NGF) by cessation of cell division and by expression of several properties resembling those of differentiated sympathetic neurons. Within minutes after addition, NGF rapidly stimulates transient expression of c-fos. To investigate the possible role of c-fos in NGF-induced differentiation, activated mouse c-fos genes were introduced into PC-12 cells by electroporation. Constitutive expression of fos inhibited NGF-induced differentiation although transfected cells harbored intact NGF receptors. Dibutyryl cyclic AMP (db cAMP) and basic fibroblast growth factor (b-FGF)-induced differentiation were also inhibited. Transcriptional activation of c-fos, c-jun, and ornithine decarboxylase (ODC) by NGF was down-regulated, whereas expression of egr-1 was unaffected in PC-12 fos clones. These results suggest that deregulated expression of fos can interfere with the normal role of NGF in neuronal differentiation. PMID- 2552375 TI - TPCK and quercetin act synergistically with vanadate to increase protein-tyrosine phosphorylation in avian cells. AB - We have found four compounds that act synergistically with the phosphotyrosine phosphatase inhibitor sodium orthovanadate (Na3VO4) to greatly increase the extent of protein-tyrosine phosphorylation in both uninfected chick embryo fibroblasts (CEF) and their Rous sarcoma virus-transformed counterparts (RSV CEF). These four inhibitors fall into two categories: the chymotrypsin-specific protease inhibitors, tosyl-phenylalanine-chloromethyl ketone (TPCK) and N carbobenzoxy-1-phenylalanine-chloromethyl ketone (ZPCK); and certain bioflavonoids which can competitively inhibit ATP-binding, quercetin and phloretin. PMID- 2552377 TI - Genetic heterogeneity within Echinococcus granulosus: isolates from different hosts and geographical areas characterized with DNA probes. AB - A segment of the ribosomal RNA gene of Schistosoma mansoni and a DNA fragment specific to Echinococcus granulosus, cloned in plasmids, have been used as DNA probes to assess the extent of genetic variability within E. granulosus and some distinct strains have been identified. The DNA analysis, involving restriction endonuclease digestion and Southern blot hybridization with the probes, did not demonstrate any significant genetic variation within the U.K. horse/dog or sheep/dog strains but confirmed the distinctiveness of the two strains shown in previous studies. The sheep/dog strain was shown to be cosmopolitan in its distribution and fertile bovine material originating from the United Kingdom, Kenya, Spain and India conformed to this strain by DNA hybridization. In contrast, cattle isolates from Holland produced markedly different DNA hybridization banding profiles indicating that cattle can harbour more than one strain of E. granulosus. Similarly, it was shown that goats can harbour two different strains of E. granulosus, the sheep/dog strain and a form which infects camels. The strain of E. granulosus infecting equines in Spain and Ireland is genetically identical to that infecting horses in the United Kingdom. There is also a different strain infecting pigs in Poland and Yugoslavia. This pig/dog strain appears to be very similar genetically to the forms of E. granulosus which use camels and goats as intermediate hosts and is similar, though not identical, to the variant infecting Dutch cattle. It has been shown that E. granulosus material, fixed for a prolonged period in ethanol, or lyophilized, is amenable to DNA analysis and that it is possible to characterize the DNA of a single adult worm. PMID- 2552376 TI - The use of magnetic resonance imaging with high-resolution CT in the evaluation of facial paralysis. AB - Magnetic resonance imaging (MRI) has been widely used in the evaluation of suspected acoustic neuroma, but has not received the same attention with respect to facial paralysis. High-resolution computed tomography (HRCT) has been the radiologic test of choice to evaluate the facial nerve. The necessary HRCT projections, slices, and enhancement techniques to visualize each segment have been outlined. We have developed a radiologic protocol that uses MRI in conjunction with HRCT, applying the strengths of each to evaluate the facial nerve. We have evaluated 15 patients and have found that MRI is the better study to evaluate the brain stem/cerebellopontine angle segment of the facial nerve and better evaluates bone-soft tissue interfaces. HRCT is better in the evaluation of the intratemporal segment of the facial nerve and the assessment of the anatomic perspectives of a lesion within the temporal bone. The results are discussed and case reports illustrate the efficacy of this approach. PMID- 2552378 TI - [Bactericidal effect of daptomycin (LY 146032) compared with vancomycine and teicoplanin against gram-positive bacteria]. AB - Daptomycin (LY 146032) is a semi-synthetic antibiotic whose structure consists of a cyclic peptide with a lipophilic chain. It inhibits synthesis of the bacterial wall differently than vancomycin and teicoplanin. The bactericidal activity of daptomycin was compared with that of vancomycin and of teicoplanin with regard to 54 Gram positive strains of bacteria: (S. aureus meti S and meti R; S. epidermidis meti S and meti R; E. faecalis, E. faecium, and Corynebacterium group JK). Vancomycin and teicoplanin have the same slow and time-dependent bactericidal activity. The activity of daptomycin is different: rapid and concentration-dependent, similar to that of the aminoglycosides. In the case of the Corynebacteria, however, the activity of daptomycin is comparable to that of vancomycin and of teicoplanin slow and time-dependent. PMID- 2552379 TI - [Bacteriostatic and bactericidal study of rifabutine and clofazimine in combination against Mycobacterium avium-intracellulare and Mycobacterium xenopi]. AB - The authors have tested the sensibility to rifabutine and clofazimine of seventeen strains of M. avium and eleven strains of M. xenopi. They compared the ability of these and other antimicrobial agents ciprofloxacin, amikacin and erythromycin alone or in combinations to inhibit the growth of strains of the M. avium complex and of M. xenopi. The purpose of this investigation was also to examine the bactericidal action of these antimicrobial agents and to determine the ability of combinations to kill strains of M. avium and M. xenopi. PMID- 2552380 TI - [Value of fluconazole in the treatment of systemic yeast infection]. AB - 20 patients (18 men, 2 women), 10 of whom were HIV +, were given Fluconazole (F) for either systemic candidiasis (13 cases), histoplasmosis (1), or cryptococcosis (6). The localization of the Candida infections (12 C. albicans, 1 C. tropicalis), were: septicemic (2), urinary (7), bronchial (2), esophageal (5), uveal (1), soft tissue (2), and 1 undetermined localization but a positive serology (1). On day (d) 1, Candidiasis patients were given an initial dose of 400 mg (for septicemia) or 200 mg (other localizations) of FIV or PO, then 200 or 100 mg per d. The length of treatment lasted from 28 to 70 d. Evolution was favorable in all the patients. 4 relapses occurred after the end of treatment: at 10 d, a septicemic candidiasis (C. tropicalis) in 1 patient who had prosthetic endocarditis; and at 1 month, digestive candidiasis in 3 HIV + patients. For the patient, infected by Histoplasma capsulatum, despite a clinical improvement, urine were still positive at day 75. The patients with cryptococcosis (5 meningitidis in the AIDS patients) and renal (1) (kidney transplant) were given on the average 400 mg a d, IV or PO (mean length 8 weeks). Only 5 patients were evaluable. For 2 of the meningitis patients with other localizations, standard treatment was instituted due to the persistence of positive cultures. For the 2 other patients, the cerebrospinal fluid (1) and the urine (1) were sterilized by the 3d week. But they relapsed 1 month after the treatment stopped. For the 18 patients evaluable, clinical and biological tolerance was good except for 1 patient with transaminases rise for which fluconazole was probably the cause. PMID- 2552381 TI - [In vitro effect of itraconazole against various species of Candida]. AB - The authors compared the in vitro antifungal activity of itraconazole to that of miconazole against various species of Candida. The MIC of 88 strains of Candida (C. albicans, C. guilliermondii, C. parapsilosis, C. glabrata) was determined by the agar dilution method using casitone medium at pH 5.5-5.6 for miconazole and pH 7.2-7.4 for itraconazole. C. albicans and C. glabrata were found to be less sensitive in vitro to itraconazole than to miconazole; the contrary was observed for C. parapsilosis. The sensitivity of C. guilliermondii to these antifungal agents was similar. The MIC 50, MIC 90 and G.MIC values for itraconazole with respect to 42 strains of C. albicans was 0.10, 1.4 and 0.138 micrograms ml-1, respectively; the MIC 90 and G.MIC values for miconazole were 0.5 and 0.021 micrograms ml-1, respectively. The effect of itraconazole on the ultrastructure of C. albicans yeast cells and spheroplasts was studied by scanning electron microscopy. This triazole molecule modified the cell wall of C. albicans, caused cell stretching and provoked defective separation between mother and daughter cells. Itraconazole altered the cytoplasmic membrane of the spheroplasts causing them to have a "spongy" appearance. Yeast cells treated with itraconazole appeared to liberate their spheroplast with difficulty. PMID- 2552382 TI - [Features of the modulation of afferent reactions by agonists and antagonists of opiate receptors and competitive stimulation of the skin under conditions of changing barometric pressure]. AB - Afferent body reactions were studied under conditions of changes of atmospheric pressure in electrostimulation of skin zones and use of various agonists and antagonists of opiate receptors. The results suggest maintenance of a sufficiently high therapeutic efficacy of transcutaneous electrostimulation at levels of up to 5.5 km above sea level and emphasize the leading role of endogenous ligands of opiate receptors in realization of the analgesic effects of competitive skin stimulation. The results of the study also provide evidence of a high analgesic activity of dalargin, a synthetic analogue of leu-enkephalin, which persisted at a height of up to 7.5 km above sea level. Increasing the resistance of the experimental animals to acute high-altitude hypoxia, this agent facilitates the development of analgesic reactions in electrostimulation of the skin. PMID- 2552383 TI - [Accumulation of 99m-technetium-pyrophosphate and the level of cyclic nucleotides in the myocardium during its adaptation to stress damage]. AB - It was shown in experiments on albino rats that their adaptation by a series of "trainings" or a course of treatment with Rhodida rosea extract produced a marked cardioprotective effect during emotional-pain stress. Adaptation of rats to stress factors induced simultaneously regular changes of the cyclic nucleotide content in the myocardium, thus contributing to reduction of the adrenoreactivity of the heart and the degree of the stress damage to the myocardium. PMID- 2552384 TI - Physiologic and metabolic aspects of anticonvulsants. AB - A great number of anticonvulsants are available for treating these different types of epilepsy. Therapeutic drug monitoring has been favored as the method for controlling drug concentrations in the plasma and preventing untoward effects. When these anticonvulsants are prescribed to treat epilepsy in children, careful monitoring is most important because drug metabolism varies depending on maturation and development of body functions. Molecular approaches are also important to elucidate the effectiveness of the drugs for treatment of different seizure disorders and should contribute to a better understanding of body functions. PMID- 2552385 TI - Pediatric uses of opioids. AB - It is evident that opioids are underused in infants and children, mainly owing to the erroneous belief that long-term adverse effects may result from effective use, coupled with the difficulties infants and children have in verbalizing their feelings of pain. Sufficient data exist to date on the safe use of opioids in the pediatric age group to allow pediatricians to incorporate them in a variety of protocols. Moreover, unlike most drugs in clinical use, opioids have a specific, safe antidote that can reverse their toxic effects promptly and effectively. PMID- 2552387 TI - IGF binding proteins in growth-retarded children with chronic renal failure. AB - Changes in the normal hormonal regulation of linear growth which lead to growth failure in children with chronic renal failure (CRF) are not well understood. Previous studies indicate that serum levels of growth hormone and IGF-I and II are normal or elevated in this population; and that serum levels of poorly defined inhibitors of IGF action are increased. Using a recently developed RIA for the 25-kD IGF-binding protein (IGF-BP25), we report significant elevations of this protein in children with CRF when compared to age- and sex-matched controls. IGF-BP25 levels correlate positively with IGF-binding activity in both populations, indicating that the RIA reflects levels of bioactive protein. Although the variation of serum IGF-BP25 with chronologic age and IGF levels are preserved in CRF, the quantitative interrelationships are disrupted. Levels of the 53-kD IGF-binding protein, an IGF-binding protein derived from the high mol wt IGF complex, were also found to be elevated in the CRF population and, unlike in the control population, did not vary with age or IGF levels. IGF-BP25 has been shown to inhibit IGF mitogenic action in vitro. Our finding of elevated levels of IGF-BP25 in children with CRF suggests that this protein may play a role in the growth retardation of pediatric chronic renal failure. The significance of the elevated IGF-BP53 levels in CRF remains uncertain. PMID- 2552386 TI - Colorectal adenocarcinoma in childhood and adolescent. Report of 11 cases and review of the literature. AB - Eleven new cases of colorectal carcinoma in children (0-15 years of age) are added to the 212 cases already described in the literature. Mucinous adenocarcinoma was the most frequent histological type. Acute abdominal obstruction was the most frequent presentation. A radiologic diagnosis of malignancy was made in only six of eleven cases. The interval between onset of clinical symptoms and diagnosis ranged from two days to one year. Duration of symptoms, pathologic findings, stage, and prognosis differed markedly as compared with adult colorectal carcinomas. In spite of its rarity, colorectal malignancies may occur in children. PMID- 2552389 TI - Varicella-zoster virus infections: chronic disease in the immunocompromised host: evidence for persistent excretion of virus. PMID- 2552388 TI - Hemodynamic and neuroendocrine adaptations of the preterm lamb left ventricle to acutely increased afterload. AB - The birth process is associated with dramatic alterations of left ventricular (LV) volume loading, pressure loading, and contractile state. The preterm LV has considerable volume loading reserve. We have assessed neuroendocrine and related hemodynamic responses of the preterm lamb LV at 0.8 gestation during acute pressure loading within the first 2-4 h after birth. We measured plasma catecholamines and hemodynamic and cineangiocardiographic parameters of LV pump performance and contractility at basal levels and during rapid LV pressure loading by partial balloon obstruction of the ascending aorta before and after propranolol. A relatively high level of propranolol (3 mg/kg) was required to produce beta-adrenoceptor blockade associated with reduction of heart rate and blood pressure, but after atrial pacing there was no detectable difference of basal LV pump performance or contractility at comparable heart rate, preload, and afterload. The LV pump performance was maintained and plasma catecholamines and LV contractility were increased when aortic systolic pressure was augmented 60% over baseline. The increased contractile state at greater afterload was minimally blunted by propranolol. Thus the preterm LV is relatively hypercontractile soon after birth and is capable of an integrated augmentation of pump performance and contractile state during pressure loading. These findings are relevant to the maintenance of adequate LV performance and successful adaptation to the acute alterations of afterload associated with the transitional circulation at birth. PMID- 2552390 TI - Fatal Coxsackie B1 virus infection in neonates. PMID- 2552391 TI - Difficulty in determining varicella-zoster immune status in pregnant women. PMID- 2552392 TI - Prematurity and risk of herpes simplex infection. PMID- 2552393 TI - Internationally adopted children and cytomegalovirus. PMID- 2552394 TI - [Post-transfusion cytomegalovirus infection in premature infants weighing less than 1,500 g]. AB - The incidence of cytomegalovirus (CMV) infection among 107 low birth weight transfused infants (birth weight less than or equal to 1,500 g) admitted to an intensive care nursery over an 18 month period was evaluated. The diagnosis of CMV infection was based on specific serologic tests (presence of IgM, increased IgG by ELISA technic) and identification of the virus in the urine. During the first 8 months, the infants received untested blood and CMV disease occurred in 8 infants out of 44 (18.2%). During the following 10 months, all transfusions performed in 63 infants were supposed to be CMV negative. However, 32 infants received untested blood due to emergency, and 5 of them developed a CMV infection (15.6%). Finally, only 31 infants received CMV negative blood without any case of CMV infection. These data clearly demonstrate that, considering the severity of the CMV disease in the premature infants, transfusions should be performed with CMV negative blood products. PMID- 2552395 TI - Mechanism of active K+ secretion by flounder urinary bladder. AB - We investigated the mechanism of active K+ transport by the urinary bladder of the winter flounder by measuring transepithelial properties in Ussing Chambers and by determining the cellular electrical potential profile using conventional microelectrodes. In the absence of transmural electrochemical potential gradients isolated bladders can exhibit a serosa-to-mucosa short circuit which is due entirely to net K+ secretion. The properties of transcellular K+ movement can be adequately described by a model which provides for active K+ uptake across the basolateral membrane via an electrogenic Na/K ATPase and K+ exit from the cell across the apical membrane down an electrochemical potential gradient via K+ channels which are blocked by mucosal barium. The conductance of the apical membranes of the transporting cells appears to be due almost solely to K+ while that of the basolateral membrane may be due largely to C1-. PMID- 2552396 TI - AlF-4 inhibits the accumulation of Ca in the endoplasmic reticulum in intact myometrial strips, but not in the rabbit ear artery. AB - AlF-4, known to be a potent modulator of G-proteins, also inhibits purified "P" type cation-transport ATPases (Missiaen et al. 1988). The aim of the present work is to find out whether AlF-4 also inhibits these ATPases in intact cells. We therefore have studied the effect of AlF-4 on the force development and on 45Ca2+ -fluxes of muscle strips from rat myometrium and rabbit ear artery. 1 mM-NaF plus 10 microM-AlCl3 induces in both tissues a tonic contraction that is completely blocked by 0.5 mM-deferoxamine. The contractile response in myometrium exceeds that of the ear artery. These contractions are independent of an activation of G proteins but their amplitude depends on [Ca2+]o. Application of AlF-4 during the loading of the endoplasmic reticulum of myometrium with Ca2+ dramatically reduces the amount of stored Ca2+ as estimated from its release induced by 1 mM-carbachol during incubation in Ca-free solution. This effect could be due to a predominant inhibitory effect of AlF-4 on the (Ca2+ + Mg2+)-ATPase of the endoplasmic reticulum. Such effect could not be established in rabbit ear artery. The increase of the fractional loss of 45Ca2+ induced by 10 microM-norepinephrine in rabbit ear artery is not inhibited by AlF-4. It is concluded that the inhibition of the endoplasmic-reticulum (Ca2+ + Mg2+)-ATPase in intact myometrial cells warns us against explaining all effects of AlF-4 on intact cells only by an activation of G-proteins.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552398 TI - Production of AMP and adenosine in the interstitial fluid compartment of the isolated perfused normoxic guinea pig heart. AB - The pathway of production of AMP and adenosine in the myocardial interstitial fluid compartment was studied in the isolated perfused normoxic guinea pig heart by collecting the transmyocardial effluent (t.m.e.) with the method of De Deckere and Ten Hoor (1977). Besides adenosine and inosine, AMP was found in t.m.e. Infusion of alpha,beta-methylene adenosine 5'-diphosphate (AOPCP), a specific inhibitor of the ecto 5'-nucleotidase, resulted in increases in t.m.e. AMP and inosine and a decrease in adenosine. Infusion of acetate producing a nearly twofold increase in myocardial AMP content did not increase the t.m.e. AMP even in the presence of AOPCP. In preparations made from 6-OH dopamine treated animals, the t.m.e. adenosine and inosine were reduced and AOPCP produced smaller increases in AMP and inosine, indicating that most if not all of the t.m.e. AMP originated from the sympathetic nerve terminals. Infusions of beta,gamma imidoadenosine and beta,gamma-methylene adenosine 5'-triphosphate (AMPPNP and AMPPCP), non-hydrolysable analogs of ATP, resulted in dose-dependent increases in the t.m.e. AMP, which were much augmented in the presence of AOPCP. AMPPNP produced similar effects in 6-OH dopamine-treated preparations. As AMPPNP and AMPPCP are good substrates of ATP pyrophosphohydrolase, these findings indicate the presence of ATP pyrophosphohydrolase in the myocardial interstitial space. PMID- 2552397 TI - Quantal stores of excitatory transmitter in nerve-muscle synapses of crayfish evaluated from high-frequency asynchronous quantal release induced by veratridine or high concentrations of potassium. AB - At single voltage-clamped opener muscle fibres of crayfish claw, 10-100 mumol/l veratridine increased within a few seconds the rate of asynchronous quantal release, n, of excitatory transmitter from n less than 1 quantum/s to n congruent to 10,000 quanta/s. Thereafter n declined exponentially either with a single, tau(2) congruent to 50 s, or with two time constants tau(1) congruent to 19 s, tau(2) congruent to 50 s. In total (t----infinity), about 0.3 million quanta were released by veratridine in a single short fibre of about 1 mm length. These values were estimated by means of the noise analysis technique and they agreed with equivalent parameters of release when 100 mmol/l K+ were used as release stimulus. Strong quantal release could be elicited only once in a single muscle by veratridine. Furthermore, the effect of veratridine on quantal release could be completely prevented by pretreatment with tetrodotoxin. In another nerve muscle preparation of crayfish, the abdominal superficial extensor muscle, up to 3 million excitatory quanta could be released by veratridine in a single fibre. In the latter muscle veratridine-induced asynchronous quantal release was strongly dependent on the extracellular concentration of Ca2+ whereas in the claw opener dependence of quantal release on extracellular Ca2+ was negligible. PMID- 2552399 TI - Calcium channel current and its sensitivity to (+) isradipine in cultured pregnant rat myometrial cells. An electrophysiological and a binding study. AB - Action of (+) isradipine (PN 200-110), a dihydropyridine derivative, was investigated on the Ca channel current in cultured cells obtained from the longitudinal layer of the pregnant rat myometrium (18-19 days of gestation). Under our experimental conditions, the inward current was attributed to L-type inward current since: (i) equimolar replacement of Ba for Ca induced an increase in the peak current and a decrease in inactivation rate; (ii) residual inward currents were recorded at the end of the pulse; (iii) membrane potential for mid inactivation was about -40 mV; (iv) the voltage dependencies of the peak current elicited from holding potentials of -40 mV and -80 mV were similar. The inward current could be reduced with nanomolar concentrations of (+) isradipine when cells were depolarized by pulses to positive potentials. This was characterized by a pronounced initial blockade, but by no increased in blockade when pulses were repeatedly applied at a frequency of 0.05 Hz. Using the double pulse procedure we confirmed that (+) isradipine did not bind to the open-state of the Ca channels. Voltage-dependence of (+) isradipine blockade was assessed by determining the steady-state availability of the Ca channels. From the shift of the inactivation curve in the presence of (+) isradipine we calculated a (K)I value of 130 pM. Scatchard analysis of the specific binding of (+)[3H] isradipine resulted in a linear plot, thereby indicating specific binding to a single class of sites with a dissociation constant Kd of about 100 pM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552400 TI - Roentgenologic appearance of a thorotrast-induced small cholangiocarcinoma in a case of thorotrastosis. An autopsy case of massive gastrointestinal bleeding of esophageal varices. AB - A small cholangiocarcinoma was detected at autopsy in a patient with thorotrastosis who died from the rupture of esophageal varices at the esophagogastric junction. Prior to the advent of recent diagnostic imaging technique, a correct antemortem diagnosis could only be obtained from tumor markers. However, the tendency for the opacity of the liver to decrease slowly with time and develop uneven trabeculation suggests that small tumors may be difficult to detect against such a non-homogeneous background. PMID- 2552401 TI - [Experimental studies of intrahepatic portal vein embolization and embolic materials]. AB - The author has developed the intrahepatic portal vein embolization for the treatment of liver cancers. The purposes of this new method are 1) extension of indications for surgery by causing compensatory hypertrophy of non-embolized lobe, 2) prevention of dissemination of the tumor cells via the portal vein, 3) causing complete ischemic necrosis of the tumor together with arterial embolization, and 4) blockade of centripetal extension of tumor thrombus. The feasibility and safety of this method were studied experimentally. Three kings of materials were prepared for embolization of the portal vein; a Lipiodol-thrombin mixture (Lp-T), a Lipiodol-fibrin adhesive mixture (Lp-F), and a mixture of Lipiodol with isobutyl-2-cyanoacrylate (Lp-IBC). The portal vein was embolized in 31 dogs, 6 with Lp-T, 14 with Lp-F, and 11 with Lp-IBC. Lp-F was used 30 to 90 seconds after preparation, which had been found to be best in an in vitro study. Lp-T and Lp-IBC could be used at any time after preparation. Embolization was done safely and reliably, except in two cases of Lp-F, by use of a balloon catheter for Lp-T or Lp-F and a coaxial catheter for Lp-IBC. Follow-up portography showed recanalization in one week in the dogs embolized with Lp-T. The obstruction was maintained for two to four weeks in the dogs embolized with Lp-F, and for four weeks in all dogs embolized with Lp-IBC. Damage in the liver was slight both macroscopically and histologically. Changes in liver function and elevation of the pressure of the portal vein were transient. The author concluded that the intrahepatic portal vein embolization was both feasible and safe when the materials tested were used, and could be an effective method for liver cancers. In clinical cases, Lp-T would be suitable for short-term occlusion, Lp-F for a moderate term, and Lp-IBC for long-term. The material should be selected with regard to the purpose. PMID- 2552402 TI - [Field localization and verification system for proton beam radiotherapy in deep seated tumors]. AB - It has been recognized that, for precise localization and verification of radiation fields, a marking drawn on the skin surface is not entirely reliable because its position relative to bony structures or internal organs is not stationary. This is especially true when treating deep-seated tumors located below the clavicle. In proton beam radiotherapy at the University of Tsukuba, we ristrict the margins around the target volume. Therefore, special care in field localization and verification is needed, and we make it a rule to take X-ray pictures on each treatment day. To accomplish this, we have developed a localize and-verify system by using fluoroscopic techniques and combining it with a real time digital image processing device. This allows for the formation of a digital image of the proton field and storage it in the optic disc. By using this system as well as verification films, filed placement errors were measured in 10 patients with deep-seated tumors. Out of a total of 190 localizations, 22% exhibited a field placement error of more than 5 mm, where patient positioning and field localization were done by using skin drawings aligned with laser beams. This result strongly suggests the necessity for daily localization and verification of radiation fields, as is being done by us, for precision in the execution of proton beam radiation therapy. PMID- 2552403 TI - [New technique of intra-arterial catheterization via the branch of left axillary artery for continuous infusion chemotherapy]. AB - A new technique of intra-arterial catheterization via the branch of left axillary artery is described. This provides relatively swift and safe insertion and long term placement of an infusion catheter through the left thoracoacrominal artery without exposure of the left axillary artery, that was required by conventional methods. With our technique, the tip of the catheter tends to enter the distal axillary artery because of oblique angulation of the trunk of the thoracoacrominal artery. Therefore, the tip of the catheter in the axillary artery must be deflected using a deflecting guidewire, so that it will advance into the subclavian artery and then down to the descending aorta. The deflection guidewire can also be of help when the catheter tip migrates into either celiac or SMA. The selective catheterization has been successful in all twenty seven patients without preliminary selective left subclavian arteriography. PMID- 2552404 TI - [Clinical studies of bronchial arterial infusion of CDDP for the treatment of lung cancer--research on the platinum concentration in the tumor and pulmonary parenchyma]. AB - To evaluate the effect of CDDP on bronchial arterial infusion (CDDP-BAI) for non small cell lung cancer platinum (Pt.) was quantified both in the plasma and the lung including parenchyma, primary tumor and lymph nodes. These data were analyzed in comparison with those of patients treated with systemic chemotherapy of CDDP (CDDP-SC). Concentration of Pt. in the plasma was measured in 8 patients treated with 100-190 mg of CDDP-BAI and 3 with 125-140 mg of CDDP-SC. In consequence, rapid decrease in value of free Pt. was noted just after the administration of CDDP with reduction by 50% resulting in 42.1 minutes after CDDP BAI and 35.0 minutes after CDDP-SC, on average. These suggested that CDDP-BAI was effective not only in the lesion infused but also in the others possibly disseminated. Surgical specimens of the lung were removed to measure concentration of Pt. 10-48 days after CDDP-BAI and 20-62 days after CDDP-SC. It measured 1.33 micrograms/g after BAI and 0.46 microgram/g after CDDP-SC in the tumor tissue on the average respectively, and 0.73 microgram/g after BAI and 0.47 microgram/g after SC in the normal lung parenchyma. On the other hand the Pt. concentration in hilar nodes measured higher in value than the tumor, when given by BAI. However, CDDP-SC almost equally caused Pt. concentration lower in value in the tumor, lymph nodes and normal lung parenchyma. As to disappearance of Pt. from the tumor during the preoperative period, there was no difference between two methods of administering CDDP. Based on these data, CDDP-BAI provided higher concentration of Pt. in the tumor, and it appeared correlative to high density of staining on angiograms. In referring to Shimosato's Classification, histological effects of CDDP-BAI varied in grade not only in each tumor but also in the area within a tumor. The present histological-chemical analysis showed a significant relationship between histological effects and Pt. concentration, especially on CDDP-BAI. PMID- 2552405 TI - RADH, a gene of Saccharomyces cerevisiae encoding a putative DNA helicase involved in DNA repair. Characteristics of radH mutants and sequence of the gene. AB - A new type of radiation-sensitive mutant of S. cerevisiae is described. The recessive radH mutation sensitizes to the lethal effect of UV radiations haploids in the G1 but not in the G2 mitotic phase. Homozygous diploids are as sensitive as G1 haploids. The UV-induced mutagenesis is depressed, while the induction of gene conversion is increased. The mutation is believed to channel the repair of lesions engaged in the mutagenic pathway into a recombination process, successful if the events involve sister-chromatids but lethal if they involve homologous chromosomes. The sequence of the RADH gene reveals that it may code for a DNA helicase, with a Mr of 134 kDa. All the consensus domains of known DNA helicases are present. Besides these consensus regions, strong homologies with the Rep and UvrD helicases of E. coli were found. The RadH putative helicase appears to belong to the set of proteins involved in the error-prone repair mechanism, at least for UV-induced lesions, and could act in coordination with the Rev3 error prone DNA polymerase. PMID- 2552406 TI - The effect of E. coli host strain on the consensus sequence of regions of the human L1 transposon. AB - We have used highly methylation tolerant host strains to clone hyper- and hypo methylated genomic elements from different regions of the same family of long interspersed repetitive elements from human DNA, specifically the 1.8 kilobase (kb) and 1.2kb KpnI fragments from members of the L1 family of transposable elements in which respectively some 18% and 2.7% of cytosines are methylated in vivo in human spleen DNA. The consensus of the DNA sequences of the ends of 13 clones from the hypomethylated region of human L1 agreed exactly with the consensus derived previously from clones made using conventional host strains. However the sequences of 18 of our clones from the 5' end of the hypermethylated region differed significantly from the sequences of clones made using conventional hosts (P less than 0.0001). The 5' region of the 1.8kb L1 region is a CpG island which, in human somatic tissue, appears to be maintained in a highly methylated state, including methylation at sites other than CpG dinucleotides. The consensus sequence of this region also has features suggestive of a previously unrecognized open reading frame. PMID- 2552407 TI - DNA duplex with the potential to change handedness after every half a turn. AB - Polymorphic forms of the DNA duplex with long stretches of structural monotony are known. Several alternating purine-pyrimidine sequences have been shown to adopt left-handed Z-conformation. We report a DNA sequence d(CGCGCGATCGAT)n exhibiting alternating right-handed B and left-handed Z helical conformation after every half a turn. Further, this unusual conformation with change in handedness after every six base pairs was induced at physiological superhelical density. PMID- 2552408 TI - Transcriptional trans-repression by the c-myb proto-oncogene product. AB - We report that the c-myb protein binds to another site, MBS-II, in the SV40 enhancer with low affinity. In co-transfection experiments with a c-myb expression plasmid, tandem repeats of the sequence containing the MBS-II site induced c-myb-dependent transcriptional repression. Results of mutational analyses of the sequence around the MBS-II site suggested that the c-myb protein represses transcription by competing with another trans-activator. These results indicate that c-myb protein can regulate transcription not only positively but also negatively. PMID- 2552409 TI - Expression of largest RNA segment and synthesis of VP1 protein of bluetongue virus in insect cells by recombinant baculovirus: association of VP1 protein with RNA polymerase activity. AB - The bluetongue virus core particles have been shown to contain an RNA-directed RNA polymerase (1). To identify the protein responsible for the virion RNA polymerase activity, the complete 3.9 Kb DNA clone representing the largest RNA segment 1 (L1) of bluetongue virus (BTV-10) was placed under control of the polyhedrin promoter of Autographa californica nuclear polyhedrosis virus (AcNPV). The derived recombinant virus was used to infect Spodoptera frugiperda cells. As demonstrated by stained polyacrylamide gel electrophoresis and by the use of bluetongue virus antibody, infected insect cells synthesized the largest protein of BTV-10 (VP1, 150 k Da). Antibody raised in rabbit to recombinant VP1 protein recognized bluetongue virus VP1 protein. The recombinant virus infected cell lysate had significantly inducible levels of RNA polymerase enzymatic activity as determined by a poly (U)-oligo (A) polymerase assay. The availability of enzymatically active bluetongue virus RNA polymerase provides a system in which we can precisely delineate the role this protein plays in the regulation of bluetongue replication. PMID- 2552410 TI - Nucleotide sequence of the cytochrome oxidase subunit I gene from rice mitochondria. PMID- 2552412 TI - The characterization of the EBV alkaline deoxyribonuclease cloned and expressed in E. coli. AB - Studies of nucleic acid homology suggest the BGLF5 open reading frame of Epstein Barr virus (EBV) encodes an alkaline deoxyribonuclease (DNase) sharing some homology with that of herpes simplex virus. We report here the expression of the BGLF5 open reading frame in E. coli and the expression of high levels of a novel alkaline DNase activity in induced cells. This alkaline DNase has been purified to apparent homogeneity as a single protein species. This is the first report of the expression of a herpesvirus coded DNase in a prokaryotic system and of the purification of the EBV DNase to demonstrable purity. It has the biochemical characteristics of a typical herpesvirus alkaline exonuclease showing a high pH optimum, an absolute requirement for Mg2+ for activity and sensitivity to high salt concentrations and polyamines. The enzyme activity was neutralized by sera from patients with nasopharyngeal carcinoma and was reactive with these sera in Western blot analysis. Thus the prokaryotic expression system described here provides an economical and efficient source of the EBV DNase for biochemical and seroepidemiological analysis. PMID- 2552411 TI - Vaccinia virus encodes a thymidylate kinase gene: sequence and transcriptional mapping. AB - The nucleotide sequence and deduced amino acid sequence of a vaccinia virus gene from the SalI F fragment are shown. The predicted polypeptide shares 42% amino acid identity over a 200 amino acid region with Saccharomyces cerevisiae thymidylate kinase (TmpK) and has low homology with herpes simplex virus deoxypyrimidine kinase. Northern blotting and S1 nuclease protection showed that the TmpK gene is transcribed early during infection and mapped the mRNA 5' end to immediately upstream of the second inframe ATG codon of the open reading frame (ORF). The encoded polypeptide is predicted to be 204 amino acids long (23.2 kD) and is almost colinear with yeast TmpK. Vaccinia virus possesses genes for TK and TmpK, separated by 57 kilobases of DNA, which are co-ordinately expressed and the encoded enzymes perform sequential steps in the same biochemical pathway. PMID- 2552413 TI - Mapping initiation sites of DNA replication in vivo using polymerase chain reaction amplification of nascent strand segments. AB - We describe a sensitive method for mapping replication initiation sites near regions of sequenced genomic DNA in vivo. It is based on selective amplification of sets of segments in purified nascent DNA strands and subsequent determination of the lengths of these strands required to include each member of the set. We demonstrate the ability of this method to accurately map a well-defined origin, that of replicating SV40 DNA. Pulse-labeled DNA from infected CV-1 cells was size fractionated on an alkaline sucrose gradient and newly-synthesized strands purified by immunoprecipitation using anti-BrdU antibodies. Three pairs of synthetic oligonucleotide primers were used to amplify three SV40 segments, using the polymerase chain reaction (PCR), at known distances from the origin. Lengths of the nascent DNA strands that allow amplification were determined by hybridization to probes homologous to the amplified segments and used to calculate position of the origin. Experiments with a mix of SV40 and human HeLa cell DNA demonstrate the applicability of the method to mapping origins present at the level of single-copy genomic sequences in mammalian cells. PMID- 2552416 TI - Deoxynucleotide-containing oligoribonucleotide duplexes: stability and susceptibility to RNase V1 and RNase H. AB - Oligoribonucleotide duplexes containing one to four 2'-deoxynucleotide residues were used as substrates for ribonuclease V1 and RNase H. Either deoxyadenosine and/or deoxythymidine were incorporated into the duplex, 5'GGCCGGAUCCGCGC3' 5'GCGCGGAUCCGGCC3' by substitution of the appropriate deoxynucleoside triphosphate into a transcription reaction with T7 RNA polymerase. The melting temperature, Tm, of the duplex (1.8 microM in strands in 50 mM NaCl) containing only ribonucleotides was 79.9 degrees C. Substitution of deoxyadenosine in both strands of the duplex lowered the Tm by 2.4 degrees C. Substitution of deoxythymidine had no measurable effect on the Tm. Comparison of RNase V1 digestion patterns of fully ribonucleotide and deoxy-substituted duplexes suggest that any distortion is localized to the site of the substitution. An oligoribonucleotide containing two deoxy residues directs specific cleavage of RNA by E. coli RNase H. Structural requirements for cleavage are proposed for RNase V1 and RNase H. PMID- 2552417 TI - Nucleotide sequence of infectious bursal disease virus genome segment A delineates two major open reading frames. PMID- 2552414 TI - Hydroxyl radical footprints reveal novel structural features around the NF I binding site in adenovirus DNA. AB - We have identified a number of as yet unknown structural abnormalities of the NF I-DNA binding site within the inverted terminal repetition of adenovirus DNA by probing it with a hydroxyl radical footprinting technique. NF I binding alters the accessibility of the deoxyribose moieties to hydroxyl radicals both at the 3' and at the 5' side of the recognition sequence 5'-TGG(N)6GCCAA-3'. A smooth bend at the 5' side of the binding sequence is already present in naked linear DNA and it is further enhanced by protein binding. This could be demonstrated not only by hydroxyl radical footprinting but also by studying the temperature dependent mobility during gel electrophoresis of DNA fragments carrying the NF I binding site at circularly permutated positions. We propose that the bent conformation at this site is responsible for facilitating protein/DNA interactions. PMID- 2552415 TI - Analysis of insertion mutants reveals two new genes in the pNRC100 gas vesicle gene cluster of Halobacterium halobium. AB - The archaebacterium, Halobacterium halobium, achieves buoyancy through synthesis of intracellular gas-filled vesicles. The plasmid-encoded gene (gvpA) specifying the major structural gas vesicle protein has previously been cloned and sequenced allowing the analysis of high-frequency mutations to the vesicle negative phenotype. Among eighteen gas vesicle mutants analyzed, four were observed to contain insertion elements 0.2 to 2 kb upstream of the structural gene. To explain the phenotype of these mutants, the upstream area was analyzed by DNA sequencing and transcriptional mapping. This analysis showed the presence of two open reading frames, gvpD and gvpE, which are of opposite transcriptional orientation to gvpA (gene order gvpA-D-E). gvpD begins 201 nucleotides from the gvpA structural gene and is 1608 nucleotides long while gvpE begins two nucleotides from the 3'-end of gvpD and is 573 nucleotides long. Primer extension analysis showed the occurrence of divergent promoters in the gvpA-gvpD intergenic region with the transcription start sites separated by 109 nucleotides. The sites of three insertion sequences in gas vesicle mutants mapped within gvpE while the fourth insertion site mapped near the N-terminal coding region of gvpD. Homology between the gvpDE gene region and a chromosomal site in a H. halobium NRC-1 derivative and in several other Halobacterium strains was identified by Southern hybridization. PMID- 2552418 TI - Complete nucleotide sequence of the human myeloperoxidase gene. PMID- 2552419 TI - Sequence of the murine myeloperoxidase (MPO) gene. PMID- 2552420 TI - Nucleotide sequence of the simian rotavirus SA11 genome segment 3. PMID- 2552421 TI - Nucleocapsid protein gene of Junin arenavirus (cDNA sequence). PMID- 2552423 TI - Experimental and clinical experience with hydroxyapatite-coated hip implants. AB - Published experimental studies have founded the principles of implant fixation using hydroxyapatite coatings. Clinical results are given on the first 100 consecutive cases of hydroxyapatite-coated total hip replacement performed since 1986. The Harris hip score rating at 1 year and longer is 98 and the specific radiologic characteristics are discussed. PMID- 2552422 TI - Complete sequence of the NS1 gene (S6 RNA) of US bluetongue virus serotype 17. PMID- 2552424 TI - Advances in nondietary management of children with atopic dermatitis. AB - This paper discusses recent advances in therapy of atopic dermatitis (AD), excluding those that include dietary management. Some of these therapies are anecdotal, experimental, or somewhat controversial. It is important to emphasize that physicians should not try what is new without first having given standard therapy a long and reasonable chance to succeed. This is important because AD does not last forever, and in many patients, mild disease heals spontaneously. PMID- 2552425 TI - Relationship between receptor occupancy at 37 degrees C and the anticonvulsant effect of flunitrazepam in rats. AB - In this investigation an attempt was made to evaluate quantitatively the relationship between benzodiazepine receptor occupancy and the anticonvulsant effect of flunitrazepam in rats. A graded measure of anticonvulsant effect was obtained on the basis of an elevation of pentylenetetrazol (PTZ) threshold concentrations. The concentration-anticonvulsant effect relationship could be described by the Emax model with an EC50 in cerebrospinal fluid of 2.9 +/- 0.8 micrograms/liter and an Emax of 227 +/- 22 mg/liter PTZ (mean +/- SE). In vitro receptor occupancy was determined in a crude brain homogenate at 0 and 37 degrees C, which yielded KD values of 2.2 +/- 0.2 and 26 +/- 2 micrograms/liter, respectively. The results obtained in both experiments were combined by focusing on free flunitrazepam concentrations. This strategy resulted in a nonlinear relationship between receptor occupancy and anticonvulsant effect of flunitrazepam, with 90% of the maximum response achieved at a degree of receptor occupancy of approximately 50% at 37 degrees C. PMID- 2552426 TI - A trial to improve the analysis of boron in biological materials. AB - For the spectrophotometric determination of boron in biological materials, two decomposition methods were used. One is the alkali fusion method, and the other is the acid decomposition method using a uniseal decomposition vessel. A long range calibration curve for the former method was obtained by using an ultrapure sulfuric acid. The alkali fusion method was applied to samples into which paraboronophenylalanine (BPA) or its hydrochloric salt (BPA.HCl) had been injected. By the acid decomposition method using a uniseal vessel, 50 mg dried liver was digested by a mixed solution of 1 ml conc. sulfuric acid and 6 ml 30% hydrogen peroxide. A problem exists, however, in that the hydrogen peroxide remaining in the decomposition solution must be eliminated without loss of boron. PMID- 2552427 TI - Prenatal diagnosis of fetal glioblastoma multiforme. AB - Intracerebral tumours of the fetus are very rare conditions, most often presenting clinically as polyhydramnios and hydrocephalus. These conditions can be diagnosed with ultrasound and clearly differentiated from hydrocephalus and other intracranial lesions. The following report is of a case of an intracerebral tumour (glioblastoma multiforme) diagnosed at 33 weeks in utero using ultrasound. The prognosis for this condition is universally poor. PMID- 2552428 TI - [Metastatic glioblastoma with transverse myelitis symptoms. Histologic and immunohistochemical analyses]. PMID- 2552429 TI - [An unusual precancerous lesion of the gallbladder]. PMID- 2552430 TI - [Long-term treatment of Cushing's disease using ketoconazole. Possibility of therapeutic escape]. AB - Five women suffering from Cushing's disease were treated with ketoconazole 800 mg per day for 2 to 28 months (mean 12.4 months). Four of them had full clinical and biochemical regression. However, after 8 months of therapy the disease failed to respond in three of these four women. Increasing the ketoconazole dosage up to 1,200 mg per day was ineffective in two patients. Such an escape phenomenon, not described until now, will restrict the use of ketoconazole in the treatment of Cushing's disease, although the drug is easy to administer and well tolerated globally and by the liver in most cases. PMID- 2552431 TI - [Papillomatosis of the biliary tract]. AB - Three new cases of papillomatosis of the biliary tract are reported. A review of the literature yielded 33 cases published since 1894, which enabled the authors to collect information on the pathological, clinical and biological characteristics of biliary papillomatosis. The contribution of imaging methods to the diagnosis and treatment is developed. The course of the disease in the published cases shows that it is severe, irrespective of the type of surgical operation performed: recurrences and initially malignant lesions are frequent, apparently benign lesions may become malignant, the death rate is high (65 per cent) and cures confirmed by a sufficiently long follow-up are rare. The indications of surgery are reviewed. Liver transplantation, which is the only way to avoid recurrence and degeneration, is discussed. PMID- 2552432 TI - [The staging of bronchial cancer]. AB - The variety of individual opinions encountered in the evaluation of lung cancer is due to the multiplicity of investigations. The intrathoracic extent is basically assessed by bronchial fibroscopy with tiered biopsies and computerized tomography. The specificity of computerized tomography in the evaluation of lymph node involvement never exceeds 75 per cent, and although this figure is higher as regards mediastinal or direct chest wall involvement, it never reaches 100 per cent. The information provided by magnetic resonance imaging is not better. Metastatic extension is evaluated by abdominal ultrasonography and computerized tomography of the brain and of the upper abdomen. Systematic radionuclide bone scanning is debatable and some other examinations must be reserved to certain histological types; this is the case with bone marrow biopsy (completed, if necessary, by monoclonal antibodies) or magnetic resonance imaging of bones in small cell carcinomas. The levels of some markers are well correlated with tumoral dissemination. PMID- 2552433 TI - Moods, misattributions and mania: an interaction of biological and psychological factors in the pathogenesis of mania. AB - Circadian rhythm dysfunction has recently been suggested to have a causal role in major depressive disorders. Against this background, experiments on circadian rhythms are outlined that yield a state of sustained sleepless activity. Such states can be brought about it seems by manipulation of the external zeitgebers to which rhythms are synchronized rather than by any alteration of the circadian clock. This can be expected to yield a disorganized rhythmic state rather than any discrete phase shifting or desynchronization of rhythms. This state it is suggested should lead to a mild dysphoria, psychomotor activation and a subtle disordering of thought form. It is proposed that these changes lead to the typical clinical picture of mania when distorted cognitively by mechanisms similar to those found in depression. There are a number of implications of this hypothesis. Firstly, mania should commonly be precipitated by similar psychosocial factors to those which precipitate depression. Secondly, similar neuroendocrine findings should be found in both depression and mania. Thirdly, similar agents should be effective in the treatment of mania and depression. Fourthly, cognitive therapy may play a significant part in the management of acute episodes of mania and reduce liability to chronicity. PMID- 2552434 TI - Rehepatectomy for treating primary liver cancer. AB - From January 1960 to June 1987, 41 cases underwent reoperation for relapse of liver cancer. Hepatectomy was performed twice in 39 cases, three and four times each in one case. The 1,3 and 5-year survival rates after the first operation were 100%, 69.58% and 54.68% respectively, while those after the second operation were 75.0%, 34.62% and 34.62% respectively. The preliminary results of rehepatectomy are thus encouraging. The indicators for reintervention, types of operation, prophylactic measures against tumor recurrence and metastases, and evaluation of rehapatectomy are discussed. PMID- 2552435 TI - Molecular dynamics effects on protein electrostatics. AB - Electrostatic calculations have been carried out on a number of structural conformers of tuna cytochrome c. Conformers were generated using molecular dynamics simulations with a range of solvent simulating, macroscopic dielectric formalisms, and one solvent model that explicitly included solvent water molecules. Structures generated using the lowest dielectric models were relatively tight, with side chains collapsed on the surface, while those from the higher dielectric models had more internal and external fluidity, with surface side chains exploring a fuller range of conformational space. The average structure generated with the explicitly solvated model corresponded most closely with the crystal structure. Individual pK values, overall titration curves, and electrostatic potential surfaces were calculated for average structures and structures along each simulation. Differences between structural conformers within each simulation give rise to substantial changes in calculated local electrostatic interactions, resulting in pK value fluctuations for individual sites in the protein that vary by 0.3-2.0 pK units from the calculated time average. These variations are due to the thermal side chain reorientations that produce fluctuations in charge site separations. Properties like overall titration curves and pH dependent stability are not as sensitive to side chain fluctuations within a simulation, but there are substantial effects between simulations due to marked differences in average side chain behavior. These findings underscore the importance of proper dielectric formalism in molecular dynamics simulations when used to generate alternate solution structures from a crystal structure, and suggest that conformers significantly removed from the average structure have altered electrostatic properties that may prove important in episodic protein properties such as catalysis. PMID- 2552436 TI - UmuC mutagenesis protein of Escherichia coli: purification and interaction with UmuD and UmuD'. AB - The introduction of a replication-inhibiting lesion into the DNA of Escherichia coli produces a marked elevation in mutation rate. The mutation pathway is a component of the induced, multigene SOS response. SOS mutagenesis is a tightly regulated process dependent on two RecA-mediated proteolytic events: cleavage of the LexA repressor to induce the UmuC and UmuD mutagenesis proteins, and cleavage of UmuD to UmuD' to activate the mutation pathway. To investigate the protein protein interactions responsible for SOS mutagenesis, we have studied the interaction of UmuC, UmuD, and UmuD'. To probe intracellular interaction, we have used immunoprecipitation techniques with antibodies against UmuC or UmuD and UmuD'. We have found that antibody to UmuC precipitates UmuD' from cell extracts, and antibody to UmuD and UmuD' precipitates UmuC. Thus we conclude that UmuC probably associates tightly with UmuD' in cells. For biochemical studies, we have purified the UmuC and UmuD' proteins to use with the previously purified UmuD. UmuC associates strongly with an affinity column of UmuD and UmuD', eluting only under strongly dissociating conditions (2 M urea or 1.5 M KSCN). UmuC also associates efficiently with UmuD or UmuD' in solution, as judged by velocity sedimentation in a glycerol gradient. The likely stoichiometry is one UmuC with a dimeric UmuD or UmuD'. From these experiments and previous work, we infer that SOS mutagenesis depends on the action of the UmuC-UmuD' complex and probably RecA to rescue a stalled DNA polymerase III holoenzyme at the DNA lesion. PMID- 2552437 TI - In vitro tyrosine phosphorylation studies on RAS proteins and calmodulin suggest that polylysine-like basic peptides or domains may be involved in interactions between insulin receptor kinase and its substrate. AB - We have investigated the in vitro tyrosine phosphorylation of the HRAS and KRAS proteins by human placental insulin receptor kinase. Purified HRAS proteins are not phosphorylated by purified insulin receptor kinase. Since the tyrosine phosphorylation of calmodulin by the insulin receptor kinase in vitro requires cofactors such as protamine and poly(L-lysine), we examined the possibility that poly(L-lysine) may also potentiate the interaction between RAS proteins and the insulin receptor. We found that purified HRAS proteins are indeed phosphorylated by purified insulin receptor kinase in the presence of poly(L-lysine). In contrast, the KRAS protein, which carries an extremely basic domain (residues 172 182, Lys-Asp-Glu-Lys6-Ser-Arg), is phosphorylated by the receptor kinase without the addition of basic proteins. We then determined whether the KRAS basic domain peptide plays a role similar to that of poly(L-lysine) and found that both the HRAS protein and calmodulin are phosphorylated by the receptor kinase in the presence of the KRAS basic domain peptide. Further examination of the role of poly(L-lysine) in potentiating tyrosine phosphorylation of the HRAS protein and calmodulin by purified insulin receptor kinase indicates that poly(L-lysine) affects the conformation of these protein substrates as well as that of the receptor kinase domain. These studies suggest that polylysine-like basic proteins or domains are required to establish the interaction between insulin receptor kinase and its substrate. PMID- 2552438 TI - Expression of rat liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase and its kinase domain in Escherichia coli. AB - The rat liver bifunctional enzyme, 6-phosphofructo-2-kinase/fructose-2,6 bisphosphatase (ATP:D-fructose-6-phosphate 2-phosphotransferase/D-fructose-2,6 bisphosphate 2-phosphohydrolase, EC 2.7.1.105/EC 3.1.3.46) and its separate kinase domain were expressed in Escherichia coli by using an expression system based on bacteriophage T7 RNA polymerase. The bifunctional enzyme (470 residues per subunit) was efficiently expressed as a protein that starts with the initiator methionine residue and ends at the carboxyl-terminal tyrosine residue. The expressed protein was purified to homogeneity by anion exchange and Blue Sepharose chromatography and had kinetic and physical properties similar to the purified rat liver enzyme, including its behavior as a dimer during gel filtration, activation of the kinase by phosphate and inhibition by alpha glycerol phosphate, and mediation of the bisphosphatase reaction by a phosphoenzyme intermediate. The expressed 6-phosphofructo-2-kinase also started with the initiator methionine but ended at residue 257. The partially purified kinase domain was catalytically active, had reduced affinities for ATP and fructose 6-phosphate compared with the kinase of the bifunctional enzyme, and had no fructose-2,6-bisphosphatase activity. The kinase domain also behaved as an oligomeric protein during gel filtration. The expression of an active kinase domain and the previous demonstration of an actively expressed bisphosphatase domain provide strong support for the hypothesis that the hepatic enzyme consists of two independent catalytic domains encoded by a fused gene. PMID- 2552439 TI - Antagonists with negative intrinsic activity at delta opioid receptors coupled to GTP-binding proteins. AB - According to classical models of drug-receptor interactions, competitive antagonists share with agonists the ability to bind to a common site on the receptor molecule. However, they are different from agonists, as they cannot trigger the "stimulus" that leads to biological responses--i.e., they lack intrinsic activity. For those receptors whose signals are transduced to effector systems by GTP-binding regulatory proteins (G proteins), a mechanistic equivalent of such a stimulus is an increased ability of agonist-bound receptor to accelerate nucleotide exchange and thus GTPase activity on the G-protein molecule. Here we show that for a member of this family of receptors (delta opioid receptors in membranes of NG108-15 neuroblastoma-glioma cells), two types of competitive antagonists can be distinguished. One type has no intrinsic activity, since it neither stimulates nor inhibits the GTPase activity of G proteins and its apparent affinity for the receptor is not altered by pertussis toxin-mediated uncoupling of receptor and G protein. The second type, however, can inhibit GTPase and thus exhibits negative intrinsic activity; its affinity for receptors is increased following uncoupling from G proteins. The existence of antagonists with negative intrinsic activity may be a general feature of several classes of neurotransmitters or hormone receptors and calls for a reevaluation of biological effects produced by competitive antagonists. PMID- 2552441 TI - gamma-Aminobutyric acid uptake by a bacterial system with neurotransmitter binding characteristics. AB - gamma-Aminobutyric acid (GABA), an amino acid, has been found in every class of living organisms. In higher organisms, GABA is a neurotransmitter and binds with high affinity and specificity to GABA receptors on neurons in a sodium independent reaction that is saturable. The role of GABA in organisms lacking nervous tissue is not known. This report describes, in a strain of Pseudomonas fluorescens, a GABA uptake system with binding characteristics like those of the GABA (type A) brain receptor. The binding was saturable and specific for GABA, was sodium-independent, was of high affinity (Km = 65 nM), and was inhibited competitively by muscimol, a potent GABA analogue. The bacterial GABA system included a homogeneous binding site, and no cooperative interaction was found between sites. To our knowledge, such a system for GABA, or other neurotransmitters, in a bacterium has not been reported. PMID- 2552440 TI - An RNA polymerase II transcription factor has an associated DNA-dependent ATPase (dATPase) activity strongly stimulated by the TATA region of promoters. AB - A transcription factor required for synthesis of accurately initiated run-off transcripts by RNA polymerase II has been purified and shown to have an associated DNA-dependent ATPase (dATPase) activity that is strongly stimulated by the TATA region of promoters. This transcription factor, designated delta, was purified more than 3000-fold from extracts of crude rat liver nuclei and has a native molecular mass of approximately 230 kDa. DNA-dependent ATPase (dATPase) and transcription activities copurify when delta is analyzed by hydrophobic interaction and ion-exchange HPLC, arguing that transcription factor delta possesses an ATPase (dATPase) activity. ATPase (dATPase) is specific for adenine nucleotides; ATP and dATP, but not CTP, UTP, or GTP, are hydrolyzed. ATPase (dATPase) is stimulated by both double-stranded and single-stranded DNAs, including pUC18, ssM13, and poly(dT); however, DNA fragments containing the TATA region of either the adenovirus 2 major late or mouse interleukin 3 promoters stimulate ATPase as much as 10-fold more effectively than DNA fragments containing nonpromoter sequences. These data suggest the intriguing possibility that delta plays a critical role in the ATP (dATP)-dependent activation of run off transcription through a direct interaction with the TATA region of promoters. PMID- 2552442 TI - Prolyl 4-hydroxylase: molecular cloning and the primary structure of the alpha subunit from chicken embryo. AB - Prolyl 4-hydroxylase (EC 1.14.11.2) is a key enzyme required for the posttranslational hydroxylation of proline residues in collagen. The enzyme is a tetramer composed of two pairs of nonidentical subunits (alpha 2 beta 2). The beta subunit is protein disulfide-isomerase, a ubiquitous enzyme found in the endoplasmic reticulum of many cell types. We report here the amino acid sequence of the alpha subunit. One cDNA clone (alpha 1) was isolated from a chicken embryo cDNA expression library in lambda gt11 by screening with anti-alpha-subunit polyclonal immunoglobulins. This alpha 1 cDNA contains an open reading frame of 1401 base pairs. A comparison of the translation of the nucleotide sequence with protein sequences obtained from the purified chicken alpha-subunit polypeptide verified that alpha 1 cDNA encoded the alpha subunit. Polymerase chain reactions were used to extend the sequence of alpha 1 cDNA toward the 5' end of alpha subunit mRNA. The mature alpha subunit is composed of 516 amino acids with a calculated molecular mass of 59,373 Da. The compiled amino acid sequence contains two potential glycosylation sites, an observation that agrees with a previous demonstration that the alpha subunit contains two N-linked oligosaccharide chains. Blot hybridization analysis of total chicken embryo RNA detected an mRNA of 3.5 kilobases, a size that closely resembles the size of the cloned cDNA. Since the expression of the alpha subunit is confined to cell types that synthesize and secrete collagens, the regulation of the synthesis of the alpha subunit may play a central role in determining the expression of prolyl 4 hydroxylase activity. PMID- 2552443 TI - Modulation of the stability of a gene-regulatory protein dimer by DNA and cAMP. AB - We describe an experimental approach to the measurement of protein subunit exchange in which biotinylated subunits mediate attachment of 35S-labeled subunits to a streptavidin column as a result of the exchange process. Application of the method to Escherichia coli catabolite activator protein (CAP) revealed that in the absence of cAMP, the dimerization equilibrium constant is 3 x 10(10) M-1, with a dimer lifetime of 300 min. Exchange of CAP subunits is accelerated at least 1000-fold by the presence of nonspecific DNA, under low ionic strength conditions. Catalysis of exchange also occurs at physiological ionic conditions. In contrast, physiological concentrations of cAMP stabilize CAP with respect to subunit exchange in either the presence or the absence of DNA. We discuss the functional implications of monomerization of gene-regulatory proteins resulting from kinetic and thermodynamic lability of their dimers. PMID- 2552444 TI - Activity of synthetic peptides from the Tat protein of human immunodeficiency virus type 1. AB - To determine which of the 86 amino acids in the Tat protein of human immunodeficiency virus type 1 (HIV-1) are important for transactivation, peptides from Tat were synthesized and their activity was measured in cells containing a chloramphenicol acetyltransferase reporter gene under control of the HIV long terminal repeat promoter. Although the Tat sequence contains arginine- and cysteine-rich stretches that are difficult to synthesize, it was possible to prepare pure peptides in good yield by using fluoren-9-ylmethoxycarbonyl (Fmoc) chemistry. A peptide containing residues 1-58 had 5-10% the activity of full length Tat. Deleting 4 amino acids from the N terminus of this peptide further reduced activity, while peptides with more extensive N-terminal deletions and peptides missing the basic region at the C terminus had no detectable activity. A peptide previously reported to transactivate, Tat-(37-62), was completely inactive in our assays. Inactive peptides were also tested as possible inhibitors of transactivation. Tat-(21-38), which contains the cysteine-rich region and can form heterodimers with intact Tat in vitro, showed inhibition at high peptide concentrations. However, this effect was not specific for Tat or for the HIV promoter, since the peptide also inhibited expression from the simian virus 40 early promoter. PMID- 2552445 TI - Z-DNA-forming sequences are spontaneous deletion hot spots. AB - Z-DNA-forming sequences are shown to elicit a biological response in Escherichia coli. Plasmids containing sequences capable of adopting the Z conformation (GC and CA/GT) are shown to be hot spots for spontaneous deletions. All the deletions involve an even number of base pairs. The distribution of the deletion events shows that the process ends when the Z-DNA-forming sequence has been reduced to a size no longer capable of adopting the Z conformation at natural superhelical density. PMID- 2552446 TI - Provirus insertion in Tpl-1, an Ets-1-related oncogene, is associated with tumor progression in Moloney murine leukemia virus-induced rat thymic lymphomas. AB - T-cell lymphomas induced in rats by Moloney murine leukemia virus acquire increasing numbers of proviruses in their genome during tumor progression in vivo and passage of tumor cells in vitro. To determine whether the proviruses progressively acquired during tumor progression play a causal role in this process, we cloned one of them from a cell line derived from the primary tumor 2772. A probe from the cellular DNA flanking the provirus was used to analyze 79 DNA samples from primary tumor tissues of 28 tumor-bearing rats and 80 DNA samples from 30 independent tumor cell lines. This analysis revealed a rearrangement in this region in the primary tumor derived from the thymus of one animal but not in a clone of the same tumor segregating in the spleen. Of the cell line DNA samples, three carried a provirus in this region. Two of these integration events had occurred independently in two clonally related sublines derived from tumor 2772, and they were followed by rapid selection in culture. On the basis of these findings this locus was named Tpl-1 (tumor progression locus 1). The Tpl-1 locus was mapped to rat chromosome 8 and to mouse chromosome 9 at a genetic distance of 1.2 +/- 0.9 centimorgans from the Ets-1 protooncogene. Although the genetic distance between Tpl-1 and Ets-1 indicates that they are different genes, analysis of Tpl-1 cDNA clones revealed that the two are closely related. PMID- 2552447 TI - Molecular cloning of a cDNA encoding a human macrophage migration inhibitory factor. AB - A cDNA encoding a human macrophage migration inhibitory factor (MIF) was isolated, through functional expression cloning in COS-1 cells, from a cDNA library prepared from a lectin-stimulated T-cell hybridoma, T-CEMB. The 115-amino acid polypeptide encoded by the MIF cDNA (p7-1) was effectively released from the transfected COS-1 cells and yielded readily detectable MIF activity in the culture supernatant despite the apparent lack of a classical protein secretory sequence. Insertional mutational analysis and elution of MIF activity from polyacrylamide gel slices demonstrated that the Mr 12,000 protein with MIF activity released by the COS-1 cells is encoded by p7-1. The p7-1 cDNA hybridized with a 700-base mRNA expressed by Con-A-stimulated lymphocytes but not unstimulated lymphocytes. The availability of the MIF cDNA clone and recombinant MIF will facilitate the analysis of the role of this lymphokine in cell-mediated immunity, immunoregulation, and inflammation. PMID- 2552448 TI - Persistent poliovirus infection of human neuroblastoma cells. AB - Two human neuroblastoma cell lines were persistently infected with poliovirus strains of all three serotypes. In persistently infected IMR-32 cells, which were studied in greatest detail, viral antigens were present in most cells, and over a 9-month period virions were found in the medium at high titers. Persistently infected cells were resistant to superinfection by Sabin 1, 2, and 3 poliovirus but sensitive to coxsackievirus B3. The viruses recovered from persistently infected cells were studied for conservation of epitopes, host cell specificity, and temperature resistance phenotype. The antigenic site 1 carried by the major capsid protein VP1 was modified on the persistent viruses of all three serotypes. This was confirmed for one virus by sequencing the corresponding genomic region in which two mutations were detected. The titers of persistent viruses were 1-3 log10 units higher on IMR-32 cells than on nonneuronal HEp-2 cells, while parental viruses had similar titers on both lines. When thermosensitive viruses were used to initiate the infection, the persistent viruses were found to be thermoresistant at 39 degrees C. Together the results indicate that the persistent infection correlated with the selection of highly mutated viral strains. Poliovirus-infected neuroblastoma cell lines thus constitute an in vitro model of chronic viral infections, which are increasingly implicated in human neural diseases. PMID- 2552450 TI - The brown adipocyte beta-adrenoceptor. PMID- 2552449 TI - Herpes simplex virus latent RNA (LAT) is not required for latent infection in the mouse. AB - During latent infection by herpes simplex virus (HSV), an abundant latency associated transcript (LAT) that is antisense to a predominant viral alpha gene (ICP0) is found localized in the nucleus of sensory neurons. We disrupted both copies of the LAT gene in the HSV-1 genome by insertion of the Escherichia coli lacZ gene under LAT promoter control. The resulting recombinant virus, RH142, does not express any detectable LAT in either latently or productively infected cells, although beta-galactosidase expression is readily detectable in sensory neurons of latently infected mice. Expression was first detectable 3 days postinoculation and continued at approximately the same level for the entire experimental period (56 days). beta-Galactosidase expression was not detectable at any time during RH142 replication in Vero cells. Thus, the kinetics of expression and cell-type specificity of the LAT gene are distinct from other HSV 1 genes that are expressed during productive growth. When latently infected trigeminal ganglia were explanted, RH142 reactivated from latency with the kinetics and an efficiency indistinguishable from the parental wild-type virus. These studies argue against any possible antisense regulatory mechanism of LAT in the regulation of viral gene expression or any role of LAT-encoded protein during the establishment or maintenance of latency in the mouse. PMID- 2552451 TI - Mammalian tissue culture growth, viral replication, and cultivation using serum replacement factor. AB - Variability, cost, and availability of fetal bovine serum are in question; thus, we examined whether using a serum replacement factor in multiple mammalian tissue cultures could support not only cell growth, but also viral replication, expression, and retention of phenotypic markers. By using a serum replacement in defined media, we demonstrated multipassage cell growth of several different cell lines and viral cultivation and replication equivalent to fetal bovine serum. Moreover, by supplementing media with a serum replacement factor we observed at least a 28% financial savings. PMID- 2552452 TI - Autonomic receptors and cyclic nucleotides of rat parotid gland following simultaneous electrical stimulation of its parasympathetic and sympathetic nerves. AB - [3H]dihydroalprenolol and [3H]quinuclidinylbenzilate binding of membranes of rat parotid gland were generally unchanged after 10, 15, 30, or 60 min of simultaneous electrical stimulation of the parasympathetic and sympathetic nerves to the gland, although stimulation of either nerve separately caused nerve specific changes in both. Concentrations of cyclic nucleotides of the gland were, however, increased significantly from levels of the unstimulated parotid gland. Cyclic GMP showed a 10-fold increase after 10 min of stimulation, whereas only a 2-fold increase in cyclic AMP was found at this time. The increases were maintained, albeit at reduced levels, at 15 and 30 min also but by 60 min both were not different from levels of the unstimulated gland. The increases induced by separate stimulation of each nerve were greater but nerve specific, and the changes induced with simultaneous stimulation tended to reflect a reigning influence of one nerve on the other. PMID- 2552453 TI - Protection of rats against pseudorabies virus infection by gamma-interferon. AB - Administration of recombinant rat gamma-interferon to rats conferred complete protection against an otherwise lethal intraperitoneal pseudorabies virus (PRV) infection. The primary target cell of the virus has been identified as the serosal cell of the peritoneum. Histologic examination showed that after infection of the underlying adventitia, the virus replicates in the myenteric and submucosal plexuses of the gastrointestinal tract; this is followed by centripetal spread to the autonomous and central nervous system. In recombinant rat gamma-interferon-treated rats, viral antigen was absent in the primary target cells and was not detected in any other organ. In interferon-treated cultures of peritoneal fibroblasts, which represent another primary target cell population in vivo, complete inhibition of PRV replication was observed. The peritoneal macrophage is not susceptible to PRV, as was shown by coculture and immunocytochemical studies. Peritoneal cells from gamma-interferon-treated rats showed enhanced major histocompatibility class II antigen expression and extrinsic antiviral activity in PRV-susceptible rat embryo fibroblasts. The results presented in this study indicate that protection by the lymphokine is likely to be based on direct inhibition of viral replication in serosal cells. PMID- 2552454 TI - An analogue of thyrotropin-releasing hormone, DN1417, decreases naloxone binding in the rat brain. AB - We explored whether thyrotropin-releasing hormone may affect opioid receptors in the rat brain. Adult male rats were intraperitoneally injected twice a day with varying doses of DN1417, a potent analogue of thyrotropin-releasing hormone, for 2 days, and opioid receptors of the brain (hypothalamus, striatum, hippocampus, midbrain, ponsmedulla, and cortex) homogenates were determined using [3H]naloxone. Intraperitoneal injection of DN1417 in a dose of 0.3 mg/100 g body wt resulted in a significant reduction in naloxone binding of the striatum as compared with the saline-injected group, whereas naloxone binding of other brain regions was not affected by DN1417. DN1417 produced a dose-dependent decrease in naloxone binding of the striatum. The affinity constant of naloxone binding was similar between the saline- and DN1417-injected groups. In vitro addition of DN1417 did not interfere with the brain naloxone binding. The distribution of 3H labeled DN1417 injected peritoneally did not differ among the brain regions. The present data imply that the opioid antagonistic action of thyrotropin-releasing hormone may be due, at least in part, to the significant decrease in the striatal opioid binding in the rat brain. PMID- 2552455 TI - Effect of dietary carbohydrates on insulin and glucagon receptors in a new model of noninsulin-dependent diabetes-SHR/N-corpulent rat. AB - A new congenic strain of rat, the SHR/N-corpulent, provides a good model for noninsulin-dependent diabetes and was used in the present study. Corpulent rats as compared to their lean littermates are obese, hyperlipidemic, and severely hyperinsulinemic, and show an age-dependent loss of glucose tolerance. Mild fasting hyperglycemia is seen only in corpulent rats fed sucrose. Since dietary sucrose is more lipogenic than starch and since insulin and glucagon are involved in lipid and carbohydrate metabolism, we studied the effect of the type of dietary carbohydrate on insulin and glucagon levels and their receptors in lean and corpulent SHR/N rats. A significant phenotypic effect was observed (corpulent greater than lean) on plasma levels of triglyceride, cholesterol, and insulin. Dietary sucrose increased these parameters in corpulent rats but not in lean rats. Insulin and glucagon binding to liver plasma membranes was lower in corpulent rats than in lean; decreases were due to fewer receptors without a significant change in affinity. Thus, in corpulent rats, in addition to hyperinsulinemia, fewer glucagon receptors and their failure to be regulated by plasma glucagon levels appear to contribute to the hyperlipidemia. Furthermore, the hyperglycemia observed in sucrose-fed corpulent rats may be due to extreme resistance to insulin despite lower plasma glucagon and fewer glucagon receptors. PMID- 2552456 TI - Administration of ethylnitrosourea to neonate hamsters increases growth and frequency of SV40-induced fibrosarcomas. AB - The in vivo interaction between the chemical carcinogen ethylnitrosourea (ENU) and the oncogenic simian virus 40 (SV40) was studied. Inbred newborn Syrian golden hamsters were injected subcutaneously with SV40 (5 x 10(6) plaque-forming units), ENU (0.5% solution, 125 or 25 mg/kg body wt), or equal mixtures of the two. Animals that received SV40 and ENU developed more tumors (100% vs 52%) within a shorter latent period (10 weeks vs 18 weeks) than animals that received SV40 alone. Animals given SV40 and ENU showed increased mortality and increased metastatic tumors (54.2% vs 30.8%) compared with those given SV40 alone. The SV40 and ENU group also exhibited multiple (greater than 10 nodules) pulmonary metastases (33.3% vs 7.7%) and metastases in multiple organs (12.5% vs 0%) compared with animals injected with SV40 alone. No difference in primary tumor size, histology, and SV40 T-antigen content was detected between SV40- and SV40/ENU-induced tumors. Four weeks after SV40 or SV40 plus ENU treatment, animals were challenged intradermally with 2.7 x 10(6) SV40-transformed hamster cells. Five weeks after challenge, 89.5% of the animals treated with SV40 and ENU and 45.4% of animals treated with SV40 developed tumors at the challenge site. Newborn animals given SV40 and ENU developed larger tumors at the challenge site (P less than 0.002) than newborns treated with SV40 alone. Thus, administration of ENU to hamsters during the neonatal stage of development produced a long lasting systemic effect that enhanced tumor development by transplanted SV40 transformed hamster cells. PMID- 2552458 TI - Radioreceptor assay for leukotriene B4 and its use for the determination of leukotriene B4 production by clotting human blood. PMID- 2552457 TI - Herpes simplex virus-specific antibodies present in tears during herpes keratitis. AB - We examined the specificity and levels of antibodies present in rabbit tears after induced infection of the rabbit cornea. Two strains of herpes simplex virus 1 (HSV) with different patterns of ocular disease were used: RE which produces stromal disease, and F which produces epithelial disease. We found that (i) IgG, IgA, and IgM antibodies were produced, (ii) the number of specific HSV antigens recognized by these antibodies was no significantly different, and (iii) postinfection (PI) timing and concentration of antibodies varied according to the disease pattern of the virus strain. The animals infected with strain F produced high levels of IgG antibodies early PI which remained constant, while IgA and IgM antibodies also increased early PI but declined after Day 16 PI. Animals infected with strain RE showed low levels of IgA and IgM antibodies which remained low. IgG antibodies increased early PI but declined at Day 16 PI. These differences in times of appearance and in amounts of antibodies in tears may be related to the clinical course of the disease. It has been shown that stromal disease has an immunopathologic basis. Inflammation, cellular infiltration of lymphocytes, and plasma cells are seen in the stroma of RE-infected animals, but these are not present in the stroma of F-infected animals. Infectious virus was not isolated from corneal explants taken from animals during the quiescent stage of the disease. The difference in pathogenicity cannot be explained in terms of specificity of tear antibodies. Even though the disease patterns were different, the number and types of HSV polypeptides recognized by both sets of tears was similar. Consequently, we believe that the immunopathology seen in the stromal disease may be due to the anatomical site of HSV antigens, rather than to differences in specificity of tear antibodies. PMID- 2552460 TI - Stimulation of arachidonate metabolism by endotoxin and the terminal complement complex C5b-8. PMID- 2552459 TI - Receptor-mediated changes in vessel tone of fresh and aged bovine cerebral arteries in vitro. PMID- 2552461 TI - In vitro synthesis of eicosanoids and cAMP by isolated hog papillary collecting duct cells. PMID- 2552462 TI - ACE-inhibitor influence on rat glomerular prostaglandin-synthesis. PMID- 2552463 TI - Different pathways initiating arachidonic acid (AA) release/thromboxane (TxA2) formation in human platelets--studies with fluoride and phorbol ester. PMID- 2552465 TI - The combination of interleukin 1 plus tumor necrosis factor causes greater generation of LTB4, thromboxanes and aggregation on human macrophages than these compounds alone. PMID- 2552464 TI - The increase in coronary flow induced by converting enzyme inhibitors is prostacyclin independent. PMID- 2552466 TI - Effect of molsidomine and its metabolites on polymorphonuclear cell function and arachidonic acid metabolism. PMID- 2552467 TI - R68070, a combined thromboxane/endoperoxide receptor antagonist (TRA) and thromboxane synthase inhibitor (TSI), prolongs the bleeding time more than aspirin in man. PMID- 2552468 TI - Eicosanoid aspects of atherosclerosis. PMID- 2552469 TI - Regulation of arachidonate release, prostaglandin synthesis, and sphincter constriction in the mammalian iris-ciliary body. AB - In the anterior segment of the eye, phosphoinositides of the iris-ciliary body are the major source of AA for PG biosynthesis. In the past several years, we have demonstrated that these phospholipids are highly enriched in AA and have an extremely high metabolic turnover. We have also discovered that their metabolism by phospholipase C, which leads to the formation of IP3 and DG and the liberation of AA, is controlled by the following Ca2+-mobilizing receptors: alpha 1 adrenergic, M3- or M4-muscarinic cholinergic, substance P, and PGs. The release of IP3, DG, and AA in the iris was also demonstrated under in vivo conditions. Furthermore, it was demonstrated that this release is associated with denervation supersensitivity and subsensitivity of the iris. Two pathways have been demonstrated in the iris through which AA can be released directly from phosphoinositides: (a) Phosphoinositides can be hydrolyzed by phospholipase C, followed by hydrolysis of DG via lipases to liberate AA, and (b) AA can be released directly from phosphoinositides via phospholipase A2. Although the evidence for a link between Ca2+-mobilizing receptors and phospholipase C, via G proteins, has been well established, the precise link between these receptors and phospholipase A2 is still unclear. Our studies indicated that PGs may be involved in regulation of contraction and relaxation of the smooth muscles of the iris by increasing IP3 accumulation and consequently Ca2+ mobilization and by elevating the level of cAMP which in turn facilitates muscle relaxation. In addition, evidence of a link between the two pathways through the Ca2+ signaling system has been suggested. In the iris, PAF was found to liberate AA from phosphoinositides through the phospholipase A2, but not the phospholipase C pathway, thus emphasizing the role of this pathway in PG synthesis in the eye. These findings demonstrate that AA release and, consequently, PG synthesis in the iris of the eye are exquisitely regulated. In some species, such as bovine, cat and dog, PGs were found to act as full Ca2+ mobilizing agonists. It is possible that PGs function to maintain muscle tone in the resting iris smooth muscle cells, in addition to their involvement in various Ca2+-dependent processes. Our studies indicate that PGs may be involved in regulation of contraction and relaxation of the smooth muscles of the iris by increasing IP3 accumulation and consequently Ca2+ mobilization and by elevating the level of cAMP which in turn facilitates muscle relaxation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2552470 TI - Novel cytochrome P450-dependent arachidonic acid metabolites and their ocular effects. AB - Our studies, as reviewed here, clearly show that in the cornea, and possibly in other ocular tissues, arachidonic acid derivatives other than the classical PGs or the more recently discovered leukotrienes can be formed, and that at least some of the AA metabolites produced via the cytochrome P450 system have potent biological activities. In fact, two of these metabolites, 12(R)-HETE (compound C) and 12-hydroxyeicosatrienoic acid (compound D; Fig. 1), appear to be much more potent than classical PGs with respect to their effect on active ion transport systems, vasodilation, or the breakdown of the blood-aqueous barrier. The fact that 12(R)-HETE was found to be a potent inhibitor of Na+-K+-ATPase activity without appreciable vascular effects or effects on barrier permeability, while compound D was found to have a potent effect on the last two parameters without any influence on the first, suggests that products of the cytochrome P450 system are capable of inducing or modulating highly specific functions. PMID- 2552471 TI - Actions of a phorbol ester, 12-O-tetradecanoylphorbol-13-acetate, on mechanical responses of vascular smooth muscle cells. PMID- 2552472 TI - The use of 31P-NMR to study smooth muscle. AB - The use of 31P NMR to study smooth muscle is hindered by low metabolite concentrations and low tissue mass. These disadvantages can be overcome due to low rates of energy utilization and the temporal stability of smooth muscle. Smooth muscle free Mg++, ADP, and the creatine analogue beta-guanidinopropionate can be studied in ways distinctive from 31P NMR of striated muscle. The pharmacological effects of naturally occurring agents such as insulin, glucose, or norepinephrine, and of NMR markers such as phenylphosphonate or 3 (trimethylsilyl)-1-propane-sulfonate on both vascular smooth muscle and perfused smooth muscles can be measured. Given sufficient collection time, 31P NMR is as useful in assessing the behavior of smooth muscle as it is of other muscle types. PMID- 2552473 TI - The role of smooth muscle tropomyosin in actomyosin system. PMID- 2552474 TI - Two types of Ca channels in mammalian intestinal smooth muscle. PMID- 2552475 TI - A calcium independent on-off switch for cardiac force generators. AB - Developed force and ATPase activity of actomyosin in cardiac muscle are regulated not only by the concentration of cytosolic calcium, but also by the state of the contractile proteins. In this study, it has been shown that cardiac actomyosin ATPase, even in the presence of adequate Ca, can exist in an inactive state. Micromolar cyclic AMP activates the ATPase, inducing substantial enzymatic activity. Both active and inactive forms of myosin can co-exist in the same cells. Mammalian hearts appear to contain a physiological mechanism for altering the response of actomyosin to optimal concentrations of Ca. PMID- 2552476 TI - Cross-bridge kinetics derived from isotonic segment dynamics in V1 and V3 containing rat cardiac muscles: effects of adrenaline and dibutyryl cyclic AMP. PMID- 2552477 TI - Central- and peripheral-type benzodiazepine receptors: similar regulation by stress and GABA receptor agonists. AB - Central- and peripheral-type benzodiazepine (BD) receptors were labelled either by 3H-flunitrazepam or 3H-Ro 5-4864 in vitro after stress and in vivo administration of GABAA and GABAB agonists. A significant increase in the density of cerebral cortex and kidney BD binding sites was observed in rats after forced swimming stress. Similar changes in both type of BD receptors were also followed when naive (stressed) and handling-habituated (unstressed) rats were used. Stress in both models was unable to change the affinity of BD receptors in cerebral cortex, but significantly lowered it in kidneys. Acute treatment of rats with muscimol (1.5 mg/kg) or (-)baclofen (5 mg/kg) resulted in marked increase in the affinity of BD binding not only in cerebral cortex but also in kidneys. After ( )baclofen treatment the number of BD binding sites was lowered in the structures studied. In a separate study mice selected according to their behavioral response to (-)baclofen (1 mg/kg) were studied. Two weeks after the selection it appeared that baclofen responders were behaviorally more "anxious" than baclofen nonresponders. The number of BD binding sites was reduced in cerebral cortex, cerebellum, heart and kidneys in baclofen responders as compared to baclofen nonresponders. In several cases the changes in peripheral BD binding sites were even more pronounced than those in central ones. The data presented here evidence that peripheral- and central-type BD receptors are regulated similarly by GABA and some models of stress. The physiological mechanisms involved in similar regulation of central- and peripheral-type BD receptors are yet unknown. PMID- 2552478 TI - Absence of effect of angiotensin II on adenylate cyclase in rat proximal renal tubular basolateral membranes. AB - 3H-Angiotensin II binding was studied in rat renal proximal tubular basolateral membranes. The characteristics of binding were found to be consistent with a receptor for angiotensin II at this site. Effects of angiotensin II on cAMP production were studied in the basolateral membranes. Angiotensin II had no effect on the time course of basal or parathyroid hormone stimulated cAMP production in basolateral membranes. Angiotensin II did not exhibit a concentration-dependent effect on cAMP production in basolateral membranes, either alone or in the presence of parathyroid hormone. The concentration response curves of cAMP production in response to parathyroid hormone in basolateral membranes were similar in the presence or in the absence of angiotensin II. We conclude that adenylate cyclase production is not linked to the receptor for angiotensin II in rat renal proximal tubular basolateral membranes. PMID- 2552479 TI - An equivalent thermal neutron fluence detector for boron neutron capture therapy. PMID- 2552480 TI - Distal penetration of proton beams: the effects of air gaps between compensating bolus and patient. PMID- 2552481 TI - In vivo analysis of nitroxide radicals injected into small animals by L-band ESR technique. PMID- 2552482 TI - [A multi-stage treatment concept for alcoholics: utilization and results of treatment]. AB - A multistage treatment model for alcoholics that is situated close to their domicile is presented in respect of organisational structure and basic therapeutic principles. Patients are compared with each other in their various stages of treatment (detoxication, counselling, therapy, aftercare) in respect of sociodemographic, alcohol-specific and psychological variables; these variables are examined with regard to their prognostic relevance for the transition to the next stage of their treatment schedule and for the success of the treatment. The resulting consequences for the treatment of alcoholics are pointed out and discussed. PMID- 2552483 TI - Separable effects of factors on activation functions in discrete and continuous models: d' and evoked potentials. AB - This article derives tests for partial-output models of information processing. A process in a partial-output model sends output, while executing, to its successors. In a stage model a process sends output to its successors only when it stops. Both models can be analyzed by manipulating factors, each affecting a different process. In discrete serial models, such factors have additive effects on reaction time (Sternberg, 1969) and sometimes on log percent correct (Schweickert, 1985). In partial-output models, such factors produce a simple pattern, the rectangle condition, in performance curves. Conditions for a partial output representation are based on decomposable structures (Krantz et al., 1971). Tests of discrete and partial-output models are simultaneously satisfied only if performance is a linear function of time. Evoked potentials (Coles et al., 1985) and d' (Dosher, 1981) illustrate results. PMID- 2552485 TI - Lack of effects of 5HT3 receptor antagonists in the social interaction and elevated plus-maze tests of anxiety in the rat. AB - The effects of three 5HT3 receptor antagonists: BRL 43964 (0.1 and 1 mg/kg, oral), GR 38032F (0.1 and 1 mg/kg, oral), and zacopride (0.01, 0.1 and 1 mg/kg, IP) were examined in low light test conditions of the social interaction test. None of the three 5HT3 receptor antagonists had a significant effect on social interaction. In contrast, in two experiments chlordiazepoxide (7.5 mg/kg) significantly increased social interaction and this effect was greatest in the unfamiliar test condition. In a third experiment, the effects of GR 38032F (0.1 and 1 mg/kg, oral) and zacopride (0.01, 0.1 and 1 mg/kg, oral) were investigated in the high light test conditions of the social interaction test; neither compound had a significant effect. In the elevated plus-maze, chlordiazepoxide (7.5 mg/kg oral or IP) significantly increased both the per cent number of entries made onto open arms and the per cent of time spent on the open arms, indicating an anxiolytic action. Zacopride (0.01, 0.1 and 1 mg/kg, oral or IP) had no significant effect in this test. The effect of the baseline rate of responding in the social interaction test on the effects of 5-HT3 antagonists is discussed. The results from the present experiment and those from other animal tests of anxiety caution against the conclusion that 5HT3 receptor antagonists are anxiolytic. PMID- 2552484 TI - Detection of benzodiazepine receptor occupancy in the human brain by positron emission tomography. AB - Benzodiazepine receptor occupancy in the brain following oral administration of clonazepam (CZP) with a dose of 30 micrograms/kg in six healthy young men and a further dose of 50 micrograms/kg in one of the subjects was estimated by carbon 11 labeled Ro15-1788 and positron emission tomography (PET). The effects of CZP on the latency of auditory event-related potentials (P300) were also studied. Overall brain 11C uptake was depressed and the % inhibition of 11C uptake in the gray matter of the brain at 30 min after [11C]Ro15-1788 injection was 15.3-23.5% (mean, n = 6) following 30 micrograms/kg CZP when compared with that in the control experiment without any previous treatment. The 11C uptake in the cerebral cortex in the subject who received both doses decreased in a dose-related manner after 30 micrograms/kg and 50 micrograms/kg CZP. The P300 latency was prolonged significantly by 30 micrograms/kg CZP [31.6 +/- 16.3 ms (mean +/- SD, n = 6), P less than 0.05]. The P300 latency in the same subject was prolonged in a dose related manner by 30 micrograms/kg and 50 micrograms/kg CZP. The technique using [11C]Ro15-1788 and PET permits comparison of the pharmacological effects with the percentage of receptor sites which benzodiazepines occupy in the human brain. P300 also seems to be useful to investigate the pharmacological effects of benzodiazepines. PMID- 2552486 TI - Diazepam and delta-9-THC: contrasting effects on the discrimination of speech sounds in nonhuman primates. AB - The adult male baboons were trained on a psychophysical procedure to discriminate five synthetic, steady-state vowel sounds (/a/, /ae/, /e/, /U/, and /c/) from one another. A pulsed train of one vowel comprised the reference stimulus during a session. Animals were trained to press a lever and release the lever only when this reference vowel sound changed to one of the comparison vowels. All animals learned the vowel discriminations rapidly and, once learned, performed the discriminations at the 95-100% correct level. The IM administration of diazepam (0.32, 1.0, 3.2, and 10.0 mg/kg) produced dose-dependent decrements in vowel discriminability. The diazepam-induced decrements in vowel discriminability were correlated with the degree of spectral frequency differences found among the different vowels, with lower vowel discriminability scores found for those vowels with smaller spectral differences from the reference vowel. In contrast, oral administration of delta-9-THC (0.32, 1.0, 3.2, and 5.6 mg/kg) produced no decrements in vowel discriminability. PMID- 2552487 TI - Effects of beta-adrenergic activity on T-wave amplitude. AB - This study addresses the hypothesis that electrocardiographic T-wave amplitude is influenced by beta-adrenergic stimulation of the heart. Beta-adrenergic activity was manipulated both pharmacologically and through behavioral challenge. Under resting conditions, 12 healthy men underwent infusion of placebo and then the beta-agonist, isoproterenol, and the beta-blocker, propranolol, in a counterbalanced, crossover design. During infusion of placebo, subjects also underwent two behavioral challenges, a structured interview and mental arithmetic. Analysis of the resting data indicated that propranolol produced a significant increase in T-wave amplitude, and isoproterenol produced significant T-wave amplitude attenuation. As previously reported, drug effects were also in evidence for heart rate. Behaviorally-induced reduction of T-wave amplitude was observed for mental arithmetic but not structured interview, which again paralleled heart rate data. Both pharmacological and behavioral data reported in this study support the hypothesis that the T-wave is significantly affected by beta-sympathetic influence on the heart. However, a nonspecific effect of heart rate change on T-wave amplitude would also account for these results. The findings are discussed in terms of their implications for the utility of T-wave amplitude in psychophysiological research. PMID- 2552489 TI - Erythema infectiosum in a primary school: investigation of an outbreak in Bury. AB - Between January and April 1987, 97 of 302 (32%) children attending a primary school in Bury, Lancashire, were affected by an exanthematous illness. The duration and extent of the episode caused considerable local interest and anxiety. Examination of 6 serum specimens obtained during the subsequent investigation demonstrated evidence of recent Human Parvovirus B19 infection in 3 children and 1 teacher. No evidence of recent rubella virus infection was found. The investigation of the episode is discussed in the context of problems which may occur in the management of similar outbreaks. PMID- 2552488 TI - Premenstrual failure of alpha-adrenergic stimulation on hypothalamus-pituitary responses in menstrual migraine. AB - In nine women suffering from menstrual migraine (MM), and in six healthy asymptomatic volunteers, plasma beta-endorphin (beta-EP), growth hormone (GH), norepinephrine (NE), and 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG) concentrations were measured in response to clonidine (0.075 mg, i.v.) stimulation. In MM patients clonidine testing was performed in both the early and the late luteal phases of the menstrual cycle. Premenstrual symptoms were prospectively evaluated in the actual cycle using the Moos Menstrual Distress Questionnaire. beta-EP (after gel chromatography) and GH were measured using radioimmunoassay. NE and MHPG were evaluated by HPLC using electrochemical detection. In both phases of the menstrual cycle clonidine significantly reduced NE and MHPG levels in MM patients and controls in a similar way. In MM patients beta-EP and GH plasma levels were stimulated by clonidine only in the early luteal phase, whereas they remained unchanged when they were stimulated in the premenstrual period. In controls the response of both hormones was not affected by the menstrual cycle. The lack of hormonal response to clonidine in MM may suggest a postsynaptic alpha 2-adrenoreceptor hyposensitivity during the premenstrual period. This demonstrates a transient vulnerability of the neuroendocrine/neurovegetative systems, and could thus be a factor facilitating the precipitation of both behavioral changes and migraine attacks. PMID- 2552490 TI - Eat for health: a nutrition and cancer control supermarket intervention. AB - The growing evidence linking dietary patterns to the incidence and prevention of chronic disease has prompted a number of prominent health and scientific agencies to publish dietary guidelines for the public. Some dietary guidelines address specific diseases, such as cancer or heart disease; others focus on overall health promotion. This situation has created a demand for nutrition education and information programs for the public. Increasingly, supermarkets are seen as potential sites for effective consumer education. Eat for Health is a joint research study by the National Cancer Institute (NCI) and Giant Food Inc., a regional supermarket chain in the Washington-Baltimore area. The study's goal was to test the feasibility of supermarkets as a site for consumer nutrition education. Eat for Health's educational focus was diet and cancer control issues in the context of dietary patterns that promote health. Particular attention was paid to reduction of fat intake and increases in consumption of dietary fiber from grains, vegetables, and fruits. Analysis of program results is currently underway; data should be available in early 1990. PMID- 2552491 TI - Plasma enteroglucagon, gastrin and peptide YY in conventional and germ-free rats refed with a fibre-free or fibre-supplemented diet. AB - Germ-free rats and conventional rats were starved and then refed with either an elemental diet (Flexical), or Flexical plus 30% kaolin, or Flexical plus 30% of a fibre mixture. Plasma levels of enteroglucagon, gastrin and peptide YY (PYY) were all significantly affected by diet. Enteroglucagon and especially PYY were significantly increased by the addition of fermentable fibre to the diet, but only in the conventional, not in the germ-free rats. Gastrin was not affected by the addition of fermentable fibre, but was increased by kaolin. Enteroglucagon and PYY were, however, both very much elevated in the germ-free animals, in which there is no proliferative response to fibre. Enteroglucagon and PYY levels were similar to those usually associated with extreme hyperproliferative states, indicating that it is unlikely that these hormones are involved in the proliferative response of the gastrointestinal tract to dietary fibre, and casting doubt on their role in other responses. PMID- 2552492 TI - The rat distal ileum has a reduced absorptive and secretory capacity compared with proximal ileum--is it to facilitate its chemosensing function? AB - Compared with the proximal ileum, the distal ileum in rat has a greatly reduced capacity for electrogenic ion secretion induced by secretagogues and for electrogenic, sodium-linked glucose transfer assessed in vitro. Similarly, in vivo, there is a great reduction in distal ileal basal fluid absorption and secretagogue-activated fluid secretion compared with proximal ileal values. This reduced absorptive and secretory capacity of the distal ileum would allow minimal changes in the concentrations of the luminal contents, facilitating its putative role as final chemosensor of the luminal contents of the small intestine. PMID- 2552493 TI - Role of chloride in hypertonicity-induced contractures of rat soleus muscle. AB - The role of chloride in contractures induced in rat soleus muscle by mannitol-, sucrose- or urea-containing hypertonic solutions was investigated. Replacement of chloride with nitrate, sulphate or perchlorate reduced the amplitude of mannitol- and sucrose-induced contractures, and the first phase of contractures induced by urea. Addition of 9-anthracene carboxylic acid had similar effects. The presence of diazepam increased the amplitude of 'hypertonic' contractures. The results show that chloride is important in the development of hypertonic contractures. PMID- 2552494 TI - The distribution of beta-adrenoceptors in human cervix. AB - The distribution of beta-adrenoceptors in sections of human cervix taken at the proliferative phase of the menstrual cycle was studied using light-microscopic autoradiography. The radioligand [125I]iodocyanopindolol (125ICYP) was used to identify specific binding sites. Moderate density of labelling by 125ICYP was seen over smooth muscle and blood vessels. The most intense labelling, however, was seen over glands and surface columnar epithelium. The association of beta adrenoceptors with glands and surface columnar epithelium suggests a possible adrenergic regulation of secretory function in the cervix. PMID- 2552496 TI - Radiation-induced pulmonary endothelial dysfunction and hydroxyproline accumulation in four strains of mice. AB - C57BL mice exposed to 14 Gy of whole-thorax irradiation develop significant histologic lung fibrosis within 52 weeks, whereas CBA and C3H mice do not exhibit substantial fibrosis during this time. The purpose of the present study was to determine whether this strain-dependent difference in radiation histopathology is associated with genetic differences in pulmonary endothelial metabolic activity or in endothelial radioresponsiveness. C57BL/6J, C57BL/10J, CBA/J, and C3H/HeJ mice were sacrificed 12 weeks after exposure to 0 or 14 Gy of 300-kV X rays to the whole thorax. Lung angiotensin converting enzyme (ACE) activity and plasminogen activator (PLA) activity were measured as indices of pulmonary endothelial function; and lung hydroxyproline (HP) content served as an index of pulmonary fibrosis. Lung ACE and PLA activities in sham-irradiated C57BL/6J and CB57BL/10J mice were only half as high as those in sham-irradiated CBA/J and C3H/HeJ mice. Exposure to 14 Gy of X rays produced a slight but nonsignificant reduction in lung ACE and PLA activity in the C57BL strains, and a significant reduction in the CBA/J and C3H/HeJ mice. Even after 14 Gy, however, lung ACE and PLA activities in CBA/J and C3H/HeJ mice were higher than those in sham irradiated C57BL/6J and C57BL/10J mice. Lung HP content in all four strains increased significantly after irradiation, but this increase was accompanied by an increase in lung wet weight. As a result, HP concentration (per milligram wet weight) remained constant or increased slightly in both C57BL strains and actually decreased in the CBA/J and C3H/HeJ mice. These data demonstrate significant genetic differences in both intrinsic pulmonary endothelial enzyme activity and endothelial radioresponsiveness among the four strains of mice. Specifically, strains prone to radiation-induced pulmonary fibrosis (C57BL/6J, C57BL/10J) exhibit only half as much lung ACE and PLA activity as do strains resistant to fibrosis (CBA and C3H). PMID- 2552495 TI - Microwave induced stimulation of 32Pi incorporation into phosphoinositides of rat brain synaptosomes. AB - Exposure of synaptosomes to microwave radiation at a power density of 10 mW/sq cm or more produced stimulation of the 32Pi-incorporation into phosphoinositides. The extent of 32Pi incorporation was found to be much more pronounced in phosphatidylinositol-4-phosphate (PIP), and phosphatidylinositol-4,5-bisphosphate (PIP2) as compared to phosphatidylinositol (PI) and phosphatidic acid (PA). Other lipids were also found to incorporate 32Pi but no significant changes in their labeling were seen after exposure to microwave radiation. Inclusion of 10 mM lithium in the medium reduced the basal labeling of PIP2, PIP and PI and increased PA labeling. Li+ also inhibited the microwave stimulated PIP2, PIP and PI labeling but had no effect on PA labeling. Calcium ionophore, A23187, inhibited the basal and microwave stimulated 32Pi labeling of PIP and PIP2, stimulated basal labeling of PA and PI and had no effect on microwave stimulated PA and PI labeling. Calcium chelator, EGTA, on the other hand, had no effect on basal labeling of PA and PI, stimulated basal PIP and PIP2 labeling but did not alter microwave stimulated labeling of these lipids. Exposure of synaptosomes to microwave radiation did not alter the chemical concentration of phosphoinositides indicating that the turnover of these lipids was altered. These results suggest that low frequency microwave radiation alter the metabolism of inositol phospholipids by enhancing their turnover and thus may affect the transmembrane signalling in the nerve endings. PMID- 2552497 TI - Radical "en Bloc" irradiation of inoperable non-small cell lung cancer in a special position of patient. PMID- 2552498 TI - [Choledocho-gastric fistula. A rare complication of a cholangiocarcinoma]. AB - A case of choledocho-gastric fistula resulting from a cholangio-carcinoma is reported. The final diagnosis was obtained by ERCP, whereas CT, barium examination, and gastroscopy initially did not allow a conclusive diagnosis. The only clinical finding was epigastric pain. Choledocho-gastric fistula is extremely rare. PMID- 2552499 TI - Proton spectroscopic imaging (Dixon method) of the liver: clinical utility. AB - The contribution of proton spectroscopic (PS) imaging to magnetic resonance (MR) imaging of the liver was assessed at 0.5 T in 55 patients with known or suspected hepatic malignancy. PS images were compared subjectively with T1- and T2-weighted spin-echo (SE) images for hepatic lesion detection and conspicuity. For hepatic metastases (n = 27), PS images were equal to T1-weighted images in lesion detection in 17 patients but showed fewer lesions in five patients and false negative results in two. When compared with T2-weighted images, PS images depicted more lesions in six patients, an equal number of lesions in 18, and fewer lesions in two. Hepatomas (n = 8) were detected with each sequence in all patients. Hepatomas were often more conspicuous on PS images than on T2-weighted images; they were of equal conspicuity on PS and T1-weighted images in most cases. Whereas fatty infiltration (n = 16) appeared on PS images as areas of low signal intensity similar to that of paraspinal muscle, it produced no detectable abnormality on either T1- or T2-weighted images. PS imaging is inferior to T1 weighted SE imaging in the detection of hepatic metastases. The major role of PS imaging at intermediate field strength is to differentiate focal fatty infiltration from hepatic metastases. PMID- 2552500 TI - [Molecular mechanisms of establishment and disruption of Epstein-Barr virus latency]. PMID- 2552501 TI - [Regulation of cell cycle by intracellular factors]. PMID- 2552502 TI - [Regulation of initiation of chromosomal replication in bacteria; role of DnaA protein and the regulation of its expression in Bacillus subtilis]. PMID- 2552503 TI - [DNA replication in mammalian cells]. PMID- 2552504 TI - Metastatic liver tumor with tumor thrombi of the portal vein. AB - Tumor thrombi of the portal vein are an important prognostic indicator for primary hepatocellular carcinoma (hepatoma), and their presence is a major criterion in differentiating intrahepatic tumors. We report four cases of metastatic liver tumors with tumor thrombi, one a gastric tumor and the others colorectal cancers, two of which were difficult to differentiate from hepatoma. The angiographic features of hepatoma are reviewed and differentiation between the two is discussed. Angiography showed the vessels of metastatic liver tumors to be of fine to mixed types. Chaotic vessels and pooling are characteristic of hepatomas, and dense tumor stains are observed only in hepatomas. The mean survival period after angiography was 3.8 months, and the prognosis was not favorable in patients having "thread-and-streak" sign on angiography. PMID- 2552505 TI - Presentation of axillary lymphadenopathy without detectable breast primary (T0 N1b breast cancer): experience at Institut Curie. AB - Between 1960 and 1985, 31 patients presented to Institut Curie with isolated axillary lymphadenopathy, of probable metastatic origin from the breast, but without clinical or radiological evidence of a breast tumor and no other primary tumor. The mean age was 54.6 years (range 39-79 years). Histological diagnosis was obtained by axillary surgery (22 cases), drill biopsy (6 cases), and cytology (3 cases). All slides were reviewed for the present study. Treatment consisted of axillary surgery followed by radiotherapy in 22 patients, radiotherapy followed by axillary surgery in 6 patients, radiotherapy followed by modified radical mastectomy in one patient, and radiotherapy alone in 2 patients. Systemic adjuvant treatment was given to 11/31 patients. The median follow-up was 9 years (range 2-26 years). Eight recurrences have appeared. Four patients recurred in the breast only (mean time to relapse: 112 months, range 63-162 months). The four other patients recurred both in breast and/or axilla (mean time to relapse: 23 months, range 7-46 months). Nine patients have developed distant metastases, of whom three also had locoregional recurrence. Among the 11 patients who had had systemic treatment, 5/11 had recurrence or metastases. The overall 5 and 10 year actuarial survival rates were 76 and 71%, respectively. The metastasis-free 5 and 10 year actuarial survival rates were 73 and 71%, respectively. Axillary metastases without clinical or radiological evidence of a primary breast tumor represents a discrete clinical entity, the prognosis of which appears to be better than that of clinical invasive breast cancer with associated lymph node involvement.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552506 TI - Sequential combination of 5-fluorouracil, cis-platinum and irradiation in unresectable non-small cell lung cancer. AB - Twenty patients with unresectable non-small cell lung carcinoma, 15 stage III and 5 stage IV (supraclavicular lymphadenopathy) were treated with a combination of three courses of chemotherapy and hypofractionated irradiation followed after 3 weeks by split-course radiotherapy. Each course was repeated every 3 weeks with the following sequence. Cis-platin (CDDP) (20 mg/m2) was given in a 20-min infusion, followed by a 2-h infusion of 5-fluorouracil (5-FU) (400 mg/m2) on days 1, 2, 5 and 6. Radiation with a dose of 3 Gy on the target volume was given on days 3 and 4, after a 2-h infusion of 5-FU (400 mg/m2). Split course of irradiation consisted of 16 Gy in 5 fractions repeated after 3 weeks interval. The objective response rate was 75%. Median follow-up was 24 months, the median survival was 14 months. The 1-year survival was 53% and the 2-year survival was 16%. PMID- 2552507 TI - Production of transgenic merino sheep by microinjection of ovine metallothionein ovine growth hormone fusion genes. AB - Seven transgenic Merino sheep have been produced by the technique of pronuclear microinjection. Two different Sheep Metallothionein-1a-Sheep Growth Hormone fusion genes were used. Four of the transgenic sheep, all of which contained the gene MTsGH5, did not express the transgene. The remaining three sheep carrying the second fusion gene, MTsGH9, expressed the gene at high levels in a variety of tissues and had elevated blood levels of sheep growth hormone. PMID- 2552508 TI - The effect of N-formyl-methionyl-leucyl-phenyl-alanine on cholinergic neurotransmission and its modulation by enkephalinase in rabbit airway smooth muscle. AB - N-formyl-methionyl-leucyl-phenylalanine (FMLP), a synthetic analogue of bacterial chemotactic peptide, may play a role in airway hyperresponsiveness, and is cleaved by neutral endopeptidase-24.11 (enkephalinase). To determine the effect of FMLP on parasympathetic contraction of airway smooth muscle and its modulation by endogenous enkephalinase, we studied isolated rabbit tracheal ring segments under isometric conditions in vitro. FMLP did not cause muscle contraction, but it potentiated the contractile response to electrical field stimulation (EFS) in a dose-dependent fashion, with the maximal increase from the baseline response being 59.8 +/- 6.2% (mean +/- S.E.M., P less than 0.001), an effect that was abolished by t-Boc-Phe-Leu-Phe-Leu-Phe, partially inhibited by pyrilamine, but not by phentolamine or [D-Pro2,D-Trp7,9]substance P. In contrast, the contractile response to administered acetylcholine was not affected by FMLP. Pretreatment of tissues with thiorphan, an enkephalinase inhibitor, further potentiated the effect of FMLP on the EFS-induced contraction. These results suggest that FMLP facilitates cholinergic neurotransmission in rabbit airway smooth muscle probably by increasing acetylcholine release, and that this effect may be modulated by enkephalinase in the airway. PMID- 2552509 TI - Pharmacological characterization of the novel helodermin/VIP receptor present in human SUP-T1 lymphoma cell membranes. AB - [Acetyl-His1]VIP stimulated adenylate cyclase with higher potency than VIP in membranes from human SUP-T1 lymphoblasts and was used as an efficient radioiodinated ligand with low non-specific binding to evaluate the relationship between receptor occupancy and adenylate cyclase activation and the possible interference of peptide T (an epitope derived from HIV envelope protein gp120). Various peptides inhibited [125I-acetyl-His1]VIP binding and activated the enzyme, their order of potency being: helodermin greater than [acetyl-His1]VIP greater than VIP = PHI = [Phe1]VIP greater than [D-Phe2]VIP = [D-Ala4]VIP = [D Phe4]PHI greater than or equal to [D-Phe4]VIP greater than [D-His1]VIP giving further support for the existence of a novel subtype of helodermin/VIP receptors. [D-Ala1]peptide T and VIP-(10-28) did not recognize the binding site and did not inhibit, even at high concentration, VIP - or VIP analogue - stimulated adenylate cyclase activities. PMID- 2552510 TI - [Retrospective analysis of 31 cases of soft tissue tumors. Comparison of MR and surgical and anatomo-pathological findings]. AB - Twenty-five patients (16 with malignant soft-tissue tumors and 9 with suspected recurrence), previously studied with conventional radiography and Computed Tomography (CT), were examined with Magnetic Resonance (MR) imaging. MR findings were surgically and anatomo-histopathologically confirmed. Six more patients with benign lesions were examined for differential diagnosis. MR capabilities are here evaluated to detect and eventually characterize the lesion, and to assess its exact location. No false positives were found, and there was only one false negative, in a case of recurrent Malignant Fibrous Histiocytoma (MFH). In another case of MFH the infiltration of the adjacent vessels and nerves was not imaged. After a careful review of all the examinations, MR imaging appears to be able to provide much information for surgical planning about tumor extent and margins, thanks to its contrast resolution between tumor and surrounding anatomical structures. Therefore, MR imaging can be reasonably employed as the examination of choice when a soft-tissue tumor is clinically suspected, because of the multi planar images it provides with and of its differential imaging parameters. PMID- 2552511 TI - [Cystic mucinous adenocarcinoma of the pancreas in a young patient. Description of a case]. PMID- 2552512 TI - Recent advances on Epstein-Barr virus infectious mononucleosis. AB - Infectious mononucleosis (IM) is an acute, self-limited lymphoproliferative disorder caused by EBV. The classic features consist of fever, malaise, easy fatigability, pharyngotonsillitis, cervical lymphadenopathy, splenomegaly, subclinical hepatitis, and atypical lymphocytosis. Symptomatic IM occurs in older children and adolescents, whereas the subclinical IM is the rule in toddlers and young children. In general the prognosis is good, even in the more prolonged and serious cases. The laboratory diagnosis relies on the demonstration of heterophil antibody in the serum. Since there is no effective therapy, management is directed toward relief of symptoms and treatment of complications. No effective ways of prevention are at hand. PMID- 2552513 TI - [4 years' evaluation of orthopedics and traumatology at an African Regional Hospital]. AB - The authors have made an evaluation of the results obtained in orthopaedic surgery and traumatology after a four year practice at the Korhogo hospital (Cote d'Ivoire). They analyse the peculiar conditions bond to the pathology, climate, environment, technical level. The incidence of post operative sepsis was 2.8 per cent, comparable to european statistics. Bed sores were exceptional, the general rate of nonunions was 2.9 per cent. The functional results were hardly analysable. However they seemed to be rather satisfactory despite the lack of post operative rehabilitation. It is concluded that an orthopaedic and traumatologic practice can be valuably conducted in Africa even far from the main centers, provided a rigorous organisation. PMID- 2552514 TI - [Pleuromediastinal lines in infants, children and adolescents--a contribution to knowledge of the normal x-ray anatomy of the mediastinal borders]. AB - The roentgen anatomy and incidence of the most frequent pleuromediastinal lines and stripes (PML) on the sagittal chest film during the first 18 years of life are analysed. It is only on involution of the thymus that the PMLs of the superior mediastinum (anterior contact line, right paratracheal stripe, arteriovenous left subclavian lines, "aortic nipple", supra-azygous posterior pleural line) are modelled out. Between the 2nd and 6th year of life they attain their normal appearance typical for adult life. The PMLs of the posterior inferior mediastinum (paraspinal lines, infra-azygous posterior pleural line) are most frequently observed between the 6th and 18th year of life. Absence of a PML as a single finding is not pathological. Displacement, widening or opacification of a PML, specially if they affect only a segment of the PML, may be indicators of a pathological process within the mediastinum itself or in the adjacent lung or pleura. PMID- 2552515 TI - [The diagnosis of pulmonary sequestration--the value of computed tomography]. AB - The complex of lung sequestration is a rare but well known entity: A segment of lung tissue located almost always in the lung bases is primarily not connected to the bronchial tree and is supplied by a systemic artery. It is very important to demonstrate these vessels to avoid accidental injuries in surgery that may cause severe bleeding. For planning surgery it is important to delineate the extent of the lesion, since it may be larger than can be seen on x-ray film and there may be secondary infectious changes of the surrounding lung tissue. CT has proved very effective in this regard. PMID- 2552516 TI - [Digital image intensifier radiography--one year's experience with a Polytron system]. AB - Since January 1988, digital image intensifier radiography has been used in the Clinic in Mannheim for DSA examinations and also in place of conventional screen/film examinations. Measurements have shown that compared with 100 mm and film/screen formats, digital radiography has poorer spatial resolution, but improved contrast resolution. The most common use of digital radiography was for examinations of the gastrointestinal tract. Using the demonstration of the mucosal fine relief pattern as a criterion of image quality, digital image intensifier radiography was able to achieve this satisfactorily. Comparison with film/screen examinations showed no loss of diagnostic information. Advantages of image intensifier radiography are reduced radiation dose, the possibility of postprocessing and economy. On the basis of 399 examinations, digital image intensifier radiography is now firmly established as part of the daily routine of the Mannheim Clinic. PMID- 2552517 TI - [The heart ventricle wall in digital subtraction angiocardiography]. AB - Digital subtraction angiography, using a time interval difference mode in relation to a suitable mask, provides a means of showing the myocardium in every phase of the cardiac cycle. The endo- and peri-cardial contours and the thickness of the myocardium in both ventricles can be demonstrated accurately. Artifacts are usually due to displacements of the heart or to tachycardia. PMID- 2552518 TI - [Quantitative computed tomographic flow measurements of aortocoronary venous bypass grafts. II. Correlation with angiographic measurements]. AB - In 26 patients with 35 aortocoronary venous bypass grafts (ACVB) quantitative invasive angiographic parameters were compared with flow parameters of contrast medium, assessed by computed tomography. Out of seven different CT-parameters, the maximum decrease of concentration of contrast medium (40 +/- 5 HE/sec; mean +/- SEM) showed a correlation with the angiographic parameter of contrast medium quantitative flow rate (7.2 +/- 0.5 cm/sec) and this correlation (r = 0.536/SEE = 23.17) is significant (p less than 0.001). We conclude that for the single patient assessment of quantitative flow in ACVB by CT with a 3-second repetition rate could be performed with limited accuracy. PMID- 2552519 TI - [Percutaneous drainage of abscesses after stomach surgery]. AB - Fifteen abscesses following various types of gastric surgery in 14 patients were treated by three different methods: 1. Multiple puncture under sonographic control, with complete drainage of the abscess. 2. The introduction of a catheter. 3. In the presence of a fistula, introduction of a catheter into the abscess through the fistula. Amongst the 14 patients, one had to undergo further surgery because of intestinal bleeding. In the other patients, there was complete healing without any complications or further surgery. PMID- 2552520 TI - [Primary lymph node metastases in malignant melanoma studied via high-resolution real-time sonography--its value and indications]. AB - To detect regional lymph node metastases 217 patients with melanoma were examined both sonographically and clinically. Metastases were found in 15 patients by means of palpation, and in 26 patients via ultrasound. Sonography was evidently superior and enabled differentiation between metastatic changes and inflammatory enlargements of lymph nodes. Because of its excellent accuracy ultrasound examination should be performed both in preoperative tumour staging and in posttherapeutic monitoring. PMID- 2552521 TI - [The value of sonography in the therapeutic supervision of induratio penis plastica (Peyronie's disease)]. AB - The article reports on 18 sonographically examined patients with clinically verified Peyronie's disease (induratio penis plastica) who underwent a basic sonographical classification according to the system of Kelami. Four weeds after the initial sonography a control study was carried out in 17 patients. 10 of these received a treatment by local instillation with Ontosein, whereas seven patients remained without treatment. In one patient an operation (Nesbit) had been performed in the meantime. 10 of the 17 examined patients presented a change of the initial findings. Seven patients out of the number of the patients who had received treatment presented a change, whereas three patients showed no change of their basic findings. The untreated group of seven patients demonstrated in three cases a change of the sonographic pattern, and four cases were unchanged. The most significant change of the sonographic appearance (treated or untreated) was an increased echogenicity of the corpora cavernosa. In some cases no plaques could be found, in one cases a new plaque developed. In two cases the initial hypoechogenic rings sign disappeared. Sonography, however, cannot prove the success of a specific therapeutic regimen, because 50% of all patients recover without treatment. PMID- 2552522 TI - [Nephrocalcinosis in childhood. Sonographic findings and differential diagnosis]. AB - Ultrasonography is the method of choice for the detection of medullary or cortical nephrocalcinosis in infancy and childhood. Compared with abdominal radiographs and computed tomography even smallest calcifications are detected more accurately. Using furosemide, ACTH, steroids or high doses of vitamin D, early forms of medullary nephrocalcinosis associated with a faint hyperechogenic rim at the margins of the renal pyramids can be diagnosed by ultrasound. Idiopathic hypercalciuria, Bartter's syndrome and renal tubular acidosis cause medullary nephrocalcinosis, whereas cortical nephrocalcinosis is the result of renal vein thrombosis; primary hyperoxaluria is associated with cortico-medullary calcifications. Due to the localisation of the nephrocalcinosis and the age distribution of the diseases, sonography merely enables us to narrow down differential diagnosis. PMID- 2552523 TI - [T2*-weighted MR images of gynecologic tumors with the FLASH sequence: the initial experiences at 1.5 T]. AB - To speed up the time taken for MR examinations, 20 patients with malignant gynaecological lesions (carcinoma of the cervix, uterus or ovary) and two operative specimen (carcinoma of the cervix) were examined by T2*-weighted flash sequences; the results were compared with T2-weighted spin echo sequences. In ten out of 20 in vivo examinations, the flash method produced a similar contrast range between tumour and normal tissues, cystic lesions being particularly well seen in gradient echo sequences. In the remaining patients, and in the surgical specimen, demarcation of the tumour was significantly poorer on the flash sequences. There was also loss of anatomic detail and an increased susceptibility to artifacts. The flash sequence as used here can therefore not be regarded as of the same diagnostic value as T2-weighted spin echo sequences in the diagnosis of gynaecological malignancies. PMID- 2552524 TI - [The progression of untreated idiopathic scoliosis in the x-ray image]. AB - The natural history of scoliosis, or lateral curvature of the spine, was followed up in 135 patients (111 girls, 24 boys) for a total average period of 52.4 months. We observed patients with a curvature of between 5 degrees and 30 degrees none of whom had been treated specifically as orthotics or with electrical stimulation or by surgery. Two groups of patients with progressive curvature were differentiated: 1) in 62.2% of the patients the curvature progressed by more than 5 degrees during the entire observation period; 2) in 36% of the patients we found an increase in curvature by more than 5 degrees within one year. Another result of our study was that idiopathic scoliosis is particularly dangerous in young patients with a "0" Risser sign. Thoracic curvatures and double major curvatures were more liable to progress than lumbar and thoracolumbar curvatures. Even a small angle of curvature in young patients must be taken seriously. PMID- 2552525 TI - [Vertebral body lesions as an interference factor in quantitative computed tomography of the lumbar spine. An analysis of 1116 standardized performed measurements]. AB - The type, frequency and location of lumbar vertebral lesions that may interfere with quantitative computed tomographic (QCT) estimation of bone mineral content were analysed in 1166 cases. All examinations were carried out with a standard protocol on the second, third and fourth vertebral body. A total of 130 lesions were identified, the majority of which included degenerative sclerosis of L4 and compression fractures of L2. Evaluation of 28 QCTs with compression fracture in one vertebral body showed that an accurate BMC value was obtained with an interendplate distance of 13 mm or more. PMID- 2552526 TI - [Experimental research on the quantitative computed tomographic prediction of the compressive strength of the thoracolumbar vertebrae]. AB - Testing 98 motion segments we investigated the possibility of a prediction of the compressive strength of thoracolumbar vertebrae by QCT. The ultimate compressive strength can be predicted from the density of the trabecular bone and from the size of the endplates--both determined by QCT--with an error of 1 kN. The increase of compressive strength in craniocaudal direction is calculated at approximately 0.3 kN per anatomical level. This variation is due to the increase of the endplate areas. PMID- 2552527 TI - [The relation of computed tomography-determined mineral content and the fracture behavior of healthy and metastatic vertebral bodies]. AB - The mineral content of 42 normal and 19 vertebrae with metastases obtained at postmortem was determined by a dual energy method and this was related to their ability to fracture. In the normal vertebrae, this relationship was expressed in the form of a logarithmic function. Amongst vertebrae with osteolytic lesions, liability to fracture increased more rapidly than mineral loss. Vertebrae with osteoblastic metastases showed increased mineral concentration with an increased liability to fracture as compared with normal vertebrae. PMID- 2552528 TI - [Lesions of the long head of the biceps--their pathogenesis and demonstration by imaging procedures (sonography, x-ray, arthrography and computed tomography)]. AB - Because of unusual anatomy and function the long head of the biceps brachii (LHB) is often subject to pathologic changes. On reviewing 354 sonographies of the shoulder (7.5 MHz), it was found that 61 (= 17%) abnormal findings of the LHB were reported such as degenerative changes accompanying impingement stadium II and III (atrophy, hypertrophy, effusion), intracapsular ruptures, acute isolated tenosynovitis and bony changes of the sulcus (bony spurs, shallow and dysplastic sulcus with subluxation of the LHB). Each abnormal finding was confirmed by x-ray and arthrography (some via CT) and compared with sonographic report. There seems to be a strikingly high percentage of rotator cuff tears connected with lesions of the LHB and the reduced filling of the sheath of the LHB, if combined lesions were apparent elsewhere in the shoulder. We consider sonography to be the method of choice in the assessment of LHB injuries; in case of verified lesion of the LHB, other pathologic conditions elsewhere in the shoulder are likely. PMID- 2552529 TI - [Fibrous dysplasia: its appearance in the magnetic resonance tomogram]. AB - Eleven patients with proven fibrous dysplasia of the skeleton (FD) were examined using magnetic resonance imaging (MRI). Almost constantly T1-weighted hypointense, T2-weighted hyperintense (cystic) as well as T1- and T2-weighted hypointense (fibrous) areas within the lesion could be demonstrated, the latter of which always exhibited either moderate or marked enhancement following intravenous application of Gd-DTPA, resulting in a typical appearance. Furthermore, T1- and T2-weighted hyperintense areas within the lesion were seen in many patients due to focal hemorrhage. No linear time correlation between the clinical symptom pain and focal areas of hemorrhage as shown by MRI, could be established. PMID- 2552530 TI - [The importance of magnetic resonance tomography in the diagnosis of cerebral infections]. AB - Amongst 1.345 MR examinations of the skull 49 patients were suspected of a cerebral infection. With a knowledge of the clinical situation, the abnormal findings were classified according to their localization, number, extent and distribution. The final diagnoses included meningitis, meningoencephalitis, and abscesses of varying etiology. CT was carried out in 29 patients; it was found that MRI was more sensitive and able to show the lesion at an earlier stage (sensitivity 90% compared with 66%). Observations of proton density and relaxation time combined with morphological criteria and clinical history reduced differential diagnoses. On the other hand, the changes due to intracranial infections lead to an inflammatory reaction that may be similar to the findings in degenerative or even tumorous cerebral lesions. This explains why twelve patients with abnormal MRI findings were erroneously diagnosed as having cerebral infections. PMID- 2552531 TI - [Subacute cerebral infarct: native and contrast medium-enhanced MRT]. AB - Twenty-five patients with clinical and CT findings indicating supratentorial cerebral infarct were prospectively studied with MR. MR examinations were performed between the 3rd and 150th day after stroke. T2-weighted axial images were compared with T1-weighted sequences after i.v. administration of Gd-DTPA. T2 weighted images demonstrated parenchymal lesions in all 25 patients. Postcontrast T1-weighted images, however, displayed pathologic contrast enhancement in only 19 areas of infarction. In 5 of the 25 patients, the use of Gd-DTPA provided additional diagnostic information by demonstrating gyral contrast enhancement as a pattern typical for ischemia. In 2 of the 25 patients, pathologic contrast enhancement was observed in an area that was isointense to the surrounding brain tissue on the T2-weighted images. In one patient, Gd-DTPA improved differentiation between areas of subacute and chronic infarction. At present, in the patient with suspected cerebral infarction, MR imaging after i.v. administration of Gd-DTPA appears recommended when clinical findings and CT are equivocal. PMID- 2552532 TI - [How certain is the diagnosis of intracerebral tumor bleeding in magnetic resonance tomography?]. AB - Three patients with intracerebral malignant tumours were examined with high-field strength MRI and CT to determine what differences might exist between a haemorrhagic tumour and a genuine intracerebral haemorrhage. Recent literature hint on characteristic signal abnormalities in comparison to pure intracerebral haematoma. However, two of our three patients showed no significant variation, in particular no delay in the evolution of methaemoglobin and no irregular haemosiderin deposition. Up to now the MRI does not seem to be the method of choice to evaluate the aetiology of intracerebral bleeding. PMID- 2552533 TI - [Early B-scan sonographic recognition of extracranial carotid atherosclerosis in patients at risk and its significance from the preventive medicine viewpoint]. AB - High-resolution 10 Mhz sonography was performed on 27 hypertensive patients, 24 diabetics, 39 hypertensive diabetics and 23 normal people in order to demonstrate asymptomatic arteriosclerotic lesions of the extra-cranial carotid artery. Compared with normal controls, diabetics and hypertensives showed increased vessel wall thickness, but only patients with hypertension showed a higher frequency of plaques and relevant carotid stenoses. The left carotid artery is involved more often. On the basis of these results, it appears that the use of B scan sonography as a preventive measure should be limited to the high-risk hypertensive population. PMID- 2552534 TI - [How strongly does the radiation burden of diagnostic x-ray studies depend on the imaging technic?]. AB - The radiation burden of an individual patient caused by a radiological examination depends strongly on the technical parameters, such as kV and mAs. As an inquiry among 150 swiss physicians showed, rather different irradiation techniques are used for the same examination. Depending on these irradiation techniques, the doses may vary by almost a factor of ten. These large variations in dose indicate that in some clinics or hospitals the radiographic techniques and the film processing are at fault. This fact has to be accounted for by future efforts of quality assurance in diagnostic radiology. PMID- 2552535 TI - [Esophageal perforation as a rare complication in the use of the Linton-Nachlas balloon catheter]. PMID- 2552536 TI - Benign scapular defects. PMID- 2552537 TI - [Spondylodiscitis in Bechterew's disease (Andersson lesion)]. PMID- 2552538 TI - [CT in echinococcosis of the lumbar spine and paravertebral structures]. PMID- 2552539 TI - Primary tracheal and bronchial adenoid cystic carcinoma (cylindroma). PMID- 2552540 TI - [Recurrence of a cutaneous, highly malignant non-Hodgkin's lymphoma with endobronchial manifestation]. PMID- 2552541 TI - MRI of transverse sinus thrombosis. PMID- 2552542 TI - [Percutaneous implantation of 2 caval filters in a double abnormality of the inferior vena cava]. PMID- 2552543 TI - [Eutopic parturition: psychoprophylaxis or extradural analgesia. Influence on the endocrine response]. AB - Prolactin, ACTH, cortisol and HGH levels have been studied on 30 pregnant women in three different periods: during the labour, at the delivery and 24 hours later. They were divided into 3 groups depending on the analgesia: I) no analgesia (n = 10); II) psychoprophylaxis (n = 10), and III) extradural analgesia (n = 10). Prolactin levels increased during delivery and 24 hours later. A significant increase of ACTH levels (p less than 0.01) was observed during the delivery in the 3 groups even though they were under hasal values 24 hours later. Cortisol increased 38% (p less than 0.01) and 52% (p less than 0.02) in II and III groups, respectively during the delivery. No difference was found with HGH. Our results suggest that endocrine response modified by labour and delivery doesn't change with different analgesia techniques. PMID- 2552545 TI - [Bronchiolo-alveolar carcinomas]. PMID- 2552544 TI - [Hepatocarcinoma with spontaneous atypical course]. AB - A case is presented of hepatocarcinoma of spontaneous long survival (close to five years), and its rarity is commented after a bibliographic review. The therapeutic importance of this finding for similar cases is emphasized. PMID- 2552546 TI - [Bronchiolo-alveolar carcinoma. Anatomo-pathological and evolutional study of a series of 52 operated cases]. AB - The postoperative outcome of bronchiolo-alveolar epithelioma (EBA) is unpredictable. We question whether a study of the anatomo-pathological structures would enable us to detect prognostic indicators. The clinical characteristics, histopathology and outcome of 52 cases of EBA were studied. 31 tumours were detected in a systematic fashion; 50 patients had excision of the tumour and in 39 cases there was no invasion of the lymphatics. 10 were of the multicentric variety and 42 were of the nodular variety and 9 of these were the centre of an inflammatory lympho-plasmocytic reaction. 20 cases revealed mucinous differentiation and 32 were non-mucinous. In the latter cases nucleo-cytoplasmic anomalies were only slightly increased or even absent. Blood vessel invasion was present in 12 cases and metastases to the air spaces in 20. The overall survival was 83% in the first year, 65% in the second year, 42% at five years and 26.5% at 10 years. The nodular lesions were compatible with a significantly better survival than the diffuse forms. Other characteristics such as whether the tumour was mucinous or not, inflammatory, showed nuclear anomalies, blood vessel invasion and airborne metastases did not seem to affect survival. PMID- 2552547 TI - [Rifabutine in the treatment of mycobacterial infections resistant to rifampicin. Preliminary results. Group for the Study and Treatment of Resistant Mycobacterial Infections (GETIM)]. AB - Three treatment protocols using rifabutine for mycobacterial infections resistant to rifampicin were prepared by a study group (GETIM) and were accepted by the ethical committee concerned. A prospective study has been carried out since April 1986. Thirty-five cases of tuberculosis with bacilli resistant to rifampicin received daily treatment with 5 to 7 mg/kg of rifabutine combined with several other drugs which were still active in vitro. Sixteen cases of M. xenopi infection occurred in individuals without apparent immune deficiency and they were treated with a daily combination of 5 to 7 mg/kg of rifabutine, 20 mg/kg of ethambutol, 3 to 5 mg/kg of isoniazid and 400 mg of ofloxacin (or 800 mg of pefloxacin). Twenty-one cases of M. avium-intracellulare infection, also in patients without any evident immune deficiency, and fifty-nine cases in patients suffering from the acquired immunodeficiency syndrome (AIDS), were treated with a similar combination in which the fluoroquinolone was replaced with 100 mg of clofazimine. During the first three months of treatment there were few major problems of toxicity or acceptability in the different combinations of drugs with the exception of three cases of leukopenia with thrombocytopenia. The proportion of negative cultures on the third month was 8 out of 24 (33%) for the cases of pulmonary tuberculosis and 10 out of 13 (77%) for the cases of M. xenopi infection, and 6 out of 11 (55%) and 9 out of 13 (69%), respectively, for infections by M. avium-intracellulare in subjects without immune deficiency and in subjects suffering from AIDS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552548 TI - [Value of repeated thoracic scanning in the surveillance of operated bronchial cancers]. AB - The authors report three cases of endothoracic tumour recurrence in patients who have been operated on for primary lung cancer revealed by CT scanning, carried out systematically in the respiratory unit to a pre-established protocol. There were no other clinical, fibre-optic, or radiological signs of these recurrences before their discovery using CT scanning. The authors discuss the value of thoracic scanning in the systematic follow up of operated cases of bronchial cancer. PMID- 2552549 TI - Direct evidence for the oxidation of palmitic acid by host-grown Mycobacterium leprae. AB - Oxidation of palmitic acid by whole-cell suspensions of Mycobacterium leprae free from host tissues was investigated using manometric techniques. After a lag period of about 6-8 h, M. leprae suspensions catalysed an active oxidation of palmitic acid, and the oxidative process (oxygen uptake) was quite sensitive to rotenone, atabrine, amytal, antimycin A and cyanide. The spectrophotometric observations indicated that the M. leprae cytochrome system, under anaerobic conditions, was reduced in the presence of palmitic acid which was completely oxidized by oxygen. These data provide direct evidence that M. leprae cells are capable of oxidizing palmitic acid, and that oxidation is mediated by the electron transport system using oxygen as the terminal electron acceptor. PMID- 2552550 TI - O2 effect on composition of chick embryonic heart and brain. AB - Heart ventricles from chick embryos incubated in 60% O2 (hyperoxia) on the 16th through the 18th days of incubation were 21% heavier than those from control embryos maintained in 21% O2 (normoxia). Heart ventricles from embyros incubated in 15% O2 (hypoxia) were 8% lighter than controls. Changes in ventricular weight were accompanied by proportional changes in protein content (21% more in hyperoxic ventricles; 8% less in hypoxic ventricles). Ventricular tissue DNA content showed a significant increase in hyperoxia. Tissue protein/DNA ratios were significantly higher in hyperoxia and lower in hypoxia. These data suggest that increased O2 availability led to hypertrophy of chick embryo ventricular cells and an increase in the level of DNA synthesis. Cytochrome oxidase activity per mg DNA was 15-25% higher in hyperoxic ventricles than in hypoxic ventricles. This result is consistent with our previous findings that alterations in O2 availability affect the O2 consumption rate of the chick emryo in ovo, and it provides direct evidence that a phenomenon repeatedly observed in vitro is of importance in vivo. In contrast to the heart, O2 availability did not affect the wet weight, protein or DNA contents, or cytochrome oxidase activity of the chick embryo brain. PMID- 2552551 TI - [Behcet's disease]. PMID- 2552553 TI - [Non-small cell bronchial cancer: experience in Liege based on a retrospective study extending over 14 years]. PMID- 2552552 TI - [Treatment of deep venous thrombosis. Comparative study of a low molecular weight heparin fragment (Fragmin) by the subcutaneous route and standard heparin by the continuous intravenous route. A multicenter study]. AB - This open, randomised multicenter trial compares the efficacy and safety of Fragmin administered subcutaneously twice daily with standard heparin administered by continuous infusion in the treatment of deep vein thrombosis (DVT). The initial dose of Fragmin is 100 U anti-Xa/kg/12 h and the further doses are adjusted according to the anti-Xa activity between 0.5 and 0.8 U/ml, 3 hours after the morning injection. The initial dose of standard heparin is 240 UI/kg/12 h. The dose adjustments are based on the daily results of APTT (1.5 - 3 times the control). Treatments efficacy are appreciated when comparing the venography performed before and after 10 days of treatment. The safety is evaluated on clinical parameters and iterative biological tests. Sixty-six patients have been included in this study. Efficacy of the two treatments is equivalent with a phlebographic improvement in respectively 79.3 p. 100 (Heparin Group) and 71.0 p. 100 (Fragmin Group) of the cases and an aggravation in 3.4 p. 100 and 6.4 p. 100 (NS) respectively. The frequency of dosage adjustments is lower and the stability of biological tests is better in the Fragmin group. In conclusion, the administration of Fragmin twice daily by subcutaneous route seems to be equivalent at least to standard heparin continuous infusion in the treatment of recent DVT. The better convenience and safety of Fragmin have to be verified on a larger panel of patients. PMID- 2552555 TI - [Possibilities and limitations of electromyography in peripheral nerve lesions]. AB - Besides the clinical examination, electromyography is the most important method in the diagnosis of peripheral nerve lesions. Together with the clinical findings it provides rather accurate information concerning the localisation, the severity and the type of a nerve lesion. However, one has to be aware of the limitations and the pitfalls of the method in order to prevent diagnostic errors. Besides a more general description of the diagnostic possibilities, the findings in traumatic nerve lesions and in entrapment neuropathies are discussed in more detail. PMID- 2552554 TI - [Diagnostic aspects of myasthenia gravis]. AB - Myasthenia gravis is described on the basis of both the physiological mechanisms of neuromuscular transmission and the current knowledge of the pathogenesis of the disease. The different laboratory tests in use are considered with regard to their diagnostic significance, in addition to the clinical features of the disease. Differential diagnosis has to include myasthenic syndromes of different origin, periodic paralysis, diseases of the peripheral motor neuron as well as certain myopathies. PMID- 2552556 TI - [Focal mononeuropathies of the lower limbs]. AB - The various clinical pictures of lower limb's mononeuropathies are described. An essential step in the diagnosis of pressure and entrapment neuropathies is to obtain an accurate description of the patient's symptoms and to ascertain whether the sensory and motor symptoms and signs correspond to the distribution of a single peripheral nerve. EMG is a useful adjunct to the physical examination and nerve conduction studies. Computer tomogram scanning may reveal bone, joint or soft tissues abnormalities responsible for the neuropathy. The attitude and the prognosis depend on the physical and functional state of damaged nerve trunks, mainly based on the classification of neuropraxia, axonotmesis and neurotmesis. PMID- 2552557 TI - [AIDS in cats harmless for people]. PMID- 2552558 TI - [Ulnar nerve lesions at the elbow--clinical aspects and surgical treatment using medial subperiosteal epicondylectomy]. AB - Ulnar neuropathy is a well recognized clinical entity caused by a variety of pathological lesions around the elbow and the sulcus olecrani. Numbness and paraesthesia in the little finger and medial half of the fourth digit are the most common complaints. Weakness of the small muscles of the hand innervated by the ulnar nerve, and a positive Tinel's sign are other features. Generally the diagnosis is confirmed by an electro-physiological study. We have described the technique of epicondylectomy and discussed the reasons why we believe it to be preferable to anterior transposition. PMID- 2552559 TI - [Palliative neurosurgical treatment of chronic pain following peripheral nerve lesions]. AB - Pathogenesis of pain after traumatic or iatrogenic lesions to peripheral nerves as well as local and conservative therapeutic possibilities are briefly reviewed. If pain subsides or in the case of relapse with establishment of a chronic pain state the therapy of choice consists in implanting a programmable neuro stimulator with the electrodes placed near the dorsal sensory roots in the cervical epidural space for the upper extremities or along the posterior columns of the medulla in the thoracic epidural space for the legs. With a success rate for long term pain control of approximately 80% this reversible method which is well tolerated by the nervous system should always be considered for deafferentation-pain (neurogenic pain). PMID- 2552560 TI - [Peripheral neuropathies in human immunodeficiency virus infection. A prospective study of 56 patients]. AB - Neurologic disorders are common in patients infected with HIV1. Judging from reported cases, the central nervous system would be more frequently involved than the peripheral nervous system (PNS). The purpose of the present prospective study was to ascertain the incidence and the nature of PNS disorders in patients infected with HIV1. Fifty-six patients [asymptomatic carriers: 5; persistent generalized lymphadenopathy (PGL): 13; AIDS related complex (ARC): 9 and acquired immunodeficiency syndrome (AIDS): 29] were submitted to clinical and laboratory investigations and 52 underwent electromyogram and nerve conduction velocity tests. CSF was examined in 29 and nerve biopsies in 28. Our findings showed that a PNS impairment was present in 50/56 patients i.e. 89 p. 100. It was mainly a moderate sensory polyneuropathy, more often at the subclinical (29/50: 58 p. 100) than at the clinical (21/50: 42 p. 100) level. It concerned asymptomatic carriers (2/5) as well as PGL (11/13), ARC (8/9) and AIDS (29/29) patients. These data show that peripheral nerves are a target for HIV1. PMID- 2552561 TI - [Familial myopathy with "cytoplasmic body" (or "spheroid") type inclusions, disclosed by respiratory insufficiency]. AB - Three patients, 72, 45 and 18 years old belonging to 3 successive generations presented with respiratory insufficiency. The clinical onset was at about 40 years of age in the two older patients and at 18 years in the youngest one. Serum enzymes of muscle origin were within normal range as well as leucocyte maltase activity. The muscle biopsy in all three patients showed numerous cytoplasmic (spheroid) bodies within type I muscle fibers. Familial cytoplasmic body myopathy with familial incidence has rarely been reported and still more rarely has been revealed by respiratory insufficiency in adult life. PMID- 2552562 TI - [Natural history of bronchiolo-alveolar cancer. Apropos of 38 cases]. AB - Thirty-eight cases of bronchioloalveolar carcinoma were selected on anatomico pathological criteria and analysed both radioclinically and histologically. At histology, this type of carcinoma is characterized chiefly by the growth of malignant cells along the alveolar walls. Cytology, which determines the nature of these cells (pneumocyte II or Clara cell), is not specific but remains important as regards phylogenesis. The course of bronchioloalveolar carcinoma gives a better understanding of its mode of propagation. Actuarial curves show that all patients with a diffuse carcinoma die very rapidly, whereas the survival rate of those who have localized lesions is 60% at 5 years. Moreover, the time required for dissemination to take place is long in assessable cases. This means that the condition should be diagnosed at an early stage on the presence of crepitant rales, on their air bronchogram at CT and on transparietal needle biopsy under CT. The chances are that early excision will result in cure. PMID- 2552563 TI - [Multifocal tuberculosis of bone. Apropos of an exceptional case]. AB - Multifocal tuberculosis of bones (MTB) is exceptional in Europe. To the few cases found in the literature the authors add another case well documented by computerized tomography and nuclear magnetic resonance and remarkable for the number of bone lesions and their coexistence with extra-skeletal lesions. The patient was a 28-year old man native of the Ivory Coast in whom the imaging techniques demonstrated no less than 19 different bone lesions plus an abscess of the iliopsoas muscle and a prevertebral pus collection. The diagnosis of MTB was confirmed by the finding of alcohol- and acid-fast bacilli at needle aspiration of the bone lesions and by the presence of folliculo-caseous Ziehl-stained granuloma on bronchial biopsies. Fourteen months after treatment with specific 4 drug therapy, the outcome is favourable. This case is exceptional by the diffusion of bone lesions and by their association with bronchial lesions due to lymph node fistulization. Modern imaging techniques (CT, NMR), clearly demonstrated the bone lesions and their extent. PMID- 2552564 TI - [X-ray diagnostic strategies in liver and pancreatic tumors]. AB - The following publication describes the diagnostic approach in different benign and malignant space-occupying lesions of the liver and pancreas. An essential prerequisite is that all imaging methods are actually available and can be made use of. While taking the cost factor into consideration, flow diagrams are worked out for haemangiomas, focal nodular hyperplasia, hepatocellular carcinoma and liver metastases, as well as for carcinoma of the pancreas and for insulinoma. PMID- 2552565 TI - [Computed tomography of pancreatic carcinoma]. AB - The ductal adenocarcinoma is the most frequent tumour of the pancreas. So far, imaging methods did not enable early diagnosis of the tumour, since in most cases it cannot be detected before it has grown to a size of 2-3 cm diameter. CT findings of the carcinoma of the pancreas as well as problems connected with the diagnosis and with staging of the tumour are presented in a concise review. PMID- 2552566 TI - [Diagnostic strategy in liver and pancreatic tumors: imaging diagnosis using MRT]. AB - Because of distinct motion artifacts MR examinations of the upper abdomen are not performed routinely. The development of rapid imaging methods (gradient echoes) makes it possible to obtain an MR image in a few seconds, hence breath-holding is possible. Consequently, images can be obtained without respiratory artifacts. Experiences with 73 patients with liver lesions are reported, using gradient echoes. The MR investigations included T2 weighted images, T1 weighted without and with contrast agent (Gd-DTPA). MR of the liver can differentiate between metastasis and haemangiomas and can help in difficult cases before resection of liver metastases. Development and improvements of artifact-reducing methods, shortening of examination times and development of oral contrast agents and further intravenous contrast agents will extend the range of MR examinations of the upper abdomen. PMID- 2552567 TI - [Malignant fibrous histiocytoma of the maxillary sinus. Clinical and therapeutic aspects]. AB - Malignant fibrous histiocytomas, or MFH, have been individualized only since the last two decades. These sarcomas which usually develop from soft tissues are no longer considered rare, except in some of their facial sinus localizations. Two chapters will be particularly developed: the paramount significance of anatomopathological investigations owing to the difficulty in establishing the diagnosis, since MFHs may be mistaken for benign processes; the treatment which is, generally speaking, the same as for bone sarcomas. PMID- 2552568 TI - Characteristics of neutral proteases present in inflamed human gingiva. AB - The existing forms of neutral proteases present in inflamed human gingiva were examined. Neutral 2 M K Cl extracts of inflamed human gingival tissue were fractionated by gel filtration on Sephacryl S-200 and the fractions were assayed for collagenase, trypsin-, chymotrypsin-, and elastase-like proteases. Apparent molecular weights of 80-85 kDa were obtained for trypsin-, chymotrypsin-, and elastase-like proteases, and 70-75 kDa for latent collagenase. Further fractionation of high molecular weight proteases on Con A-Sepharose revealed that, unlike collagenase, chymotrypsin- and elastase-like proteases, the trypsin like protease was bound by the affinity column. Native human placental type IV (basement membrane) collagen was degraded by chymotrypsin-like and elastase-like proteases but not by the trypsin-like protease. This degradation was inhibited by phenylmethyl sulfonyl fluoride and EDTA. The serine proteases also degraded efficiently denatured type I collagen. No correlation of the activities of trypsin-like protease and the other proteolytic enzymes was found in extracts of 18 individual gingival specimens. Significant correlation, however, was noted between collagenase and gelatinase. The gingival culture studies showed that, while the highest activity of the trypsin-, chymotrypsin-, and elastase-like enzymes were measured in medium during first days of the culture, collagenase and gelatinase activities increased up to the fourth day of culture and stayed high until the end of the culture. These results suggest that the neutral proteases that may participate in the periodontal tissue destruction are produced by different cell types of gingiva. PMID- 2552569 TI - Stimulatory effect of short-chain fatty acids on epithelial cell proliferation of isolated and denervated jejunal segment of the rat. AB - Involvement of the mesenteric nerve in the efferent transmission of the trophic effect of short-chain fatty acids was tested in rats. A short segment of jejunum was translocated under the skin with or without its mesenteric connection maintained. A mixture of acetic, propionic, and n-butyric acids (100, 20, and 60 mM, respectively) or 0.9% NaCl (control) was injected into the caecum via an ileostomy twice a day for 14 days. Trophic effects of short-chain fatty acids were observed in jejunal and caecal segments and in the translocated jejunal segments both with and without mesenteric connection. The results suggest that short-chain fatty acids given into the hindgut lumen can indirectly stimulate the epithelial cell proliferation of a distant intestinal segment without luminal continuation to the site of administration, and this trophic effect does not require efferent transmission by nerves in the mesentery. Blood-born mediation is likely. PMID- 2552570 TI - Testicular GnRH-receptors and direct effects of a GnRH-agonist on human testicular steroidogenesis. AB - GnRH-like substances have been isolated from interstitial fluid from the rat testis and have been found to exert direct, mainly inhibitory effects on steroidogenesis. In rat testis, specific GnRH receptors have been shown, but so far no GnRH-receptors have been isolated from the human testis. In this study testicular tissue from nine elderly men was incubated with either 3H-pregnenolone or 3H-progesterone and the steroid metabolic patterns were analyzed. In parallel incubations a GnRH-agonist was added to the incubate in concentrations of 10(-6) M or 10(-7) M. Furthermore, receptor studies were performed in tissue specimens from four of these untreated patients as well as in tissue specimens from five GnRH-agonist treated patients. No consistent effects were elicited by the addition of GnRH-agonist on the steroid metabolic patterns in vitro after three hours incubation. In the receptor studies the GnRH-agonist was bound to the testicular tissue, although the binding sites were of low affinity and high capacity, indicating a less specific kind of binding than classical receptor binding. PMID- 2552571 TI - Leukotriene b4 production by peripheral blood neutrophils in rheumatoid arthritis. AB - Leukotriene B4 (LTB4) is an activator of white blood cells (WBC) and it has been suggested that its inhibition may be useful in rheumatoid arthritis (RA). Its production by peripheral WBC has not yet been investigated. We measured LTB4 production in 105 patients with RA and compared it with 59 matched controls. C reactive protein (CRP) and ESR were measured in 90 patients and correlated with LTB4 values. Ten millilitres of blood were drawn. Separation was undertaken to obtain polymorphonuclear leukocytes (PMN) which were stimulated with calcium ionophore, and the supernatant was frozen for radioimmunoassay of LTB4. Results show that RA patients produce significantly higher levels of LTB4. It has been suggested that blockage of the cyclo-oxygenase enzyme by non-steroid anti inflammatory drugs (NSAID) leads to increased production of LT via the lipoxygenase enzyme. Twenty-one patients not taking NSAID were compared with 84 on therapy. There was no significant difference. A linear regression was used to obtain Pearson's correlation coefficients. With LTB4 and CRP, r = 0.3 (p less than 0.003). With LTB4 and ESR, r = 0.25 (p less than 0.02). Low but significant correlations with CRP and ESR were obtained. PMID- 2552572 TI - [Selective vitamin B 12 malabsorption in a 19-year-old patient]. AB - Selective malabsorption of vitamin B12, or Imerslund-Grasbeck disease, is a rare congenital condition. The normal ileal uptake of intrinsic-factor-bound vitamin B12 is deficient. Inheritance is autosomal recessive. Megaloblastic anemia generally appears in infancy and is usually accompanied by mild proteinuria. The case is presented of a 19-year-old woman with megaloblastic anemia due to selective vitamin B12 malabsorption. As far as we know this is the only patient in whom the disease appeared so late. Of special interest are also the marginally low Schilling tests of the patient's parents. Further conditions which may lead to vitamin B12 deficiency are also discussed. A hypothesis is put forward concerning the late appearance of the disease in this particular case. Pathophysiologic, nephrologic and genetic aspects of selective vitamin B12 malabsorption are briefly reviewed. PMID- 2552573 TI - [PTH-like tumor peptide and malignant hypercalcemia]. AB - Certain patients with malignant hypercalcemia display enhanced bone resorption and changes in renal tubular handling of calcium and phosphate similar to those encountered in primary hyperparathyroidism. A tumoral protein probably responsible for this syndrome has recently been described. Produced by a gene other than that of parathyroid hormone, it shows an aminoterminal homology with the physiological hormone. In some respect both proteins exert similar effects. This parathyroid hormone-analogue may also be involved in physiological regulatory functions. PMID- 2552574 TI - [Cannabis: a harmless drug?]. AB - Summarizing the literature of the past 25 years, we present a survey of the pharmacological effects of cannabis, which includes the pharmacological particularities of cannabis as a composition of substances with tetra-hydro cannabinol as main component, the experimentally proved effects on animal cells, the experimentally and clinically proved damages to the central nervous system, respiratory tract, immune system and procreative system. The survey results in a warning: brain damage and dependency can be caused by cannabis and the resulting social consequences of an increasing consumption should not be underestimated. PMID- 2552576 TI - [Comment on the editorial "Fischtran" by P.W. Straub, Schweiz. med. Wschr. 1989; 119:963-964]. PMID- 2552575 TI - [Angioedema caused by enalapril (Reniten)]. AB - Two cases of enalapril(Reniten)-induced angioedema are described. In both patients the time lag between the first manifestation of angio-edema and diagnosis was more than one year, during which several bouts of edema occurred. One patient developed life-threatening swelling of the tongue and the larynx followed by asystole and apnea. The second patient had recurrent edema of the tongue and dyspnea. In general, enalapril-induced edema is not thought to be based on immunological mechanisms. However, in both patients we found elevated titres of antinuclear antibodies, which were reversible upon cessation of enalapril medication. The possible pathomechanisms are discussed. PMID- 2552577 TI - [Radioimmunoscintigraphy with a 99mTc-labeled F(ab')2 fragment of a monoclonal antibody (HMW-MAA 225.28S) in 71 patients with malignant melanoma]. AB - In 71 patients with malignant melanoma 85 radioimmunoscintigraphies (RIS) with 99mTc-radiolabeled F(ab')2 fragments of a murine monoclonal antibody have been performed. The antibody is specific for an epitope of the HMW-MAA glycoprotein complex which is present in 90% of melanoma tissue samples. Sensitivity for RIS was 79% (73 metastatic sites out of a total of 92). 20 previously unknown lesions were found and there were also 2 false positives. Conventional staging procedures alone revealed 78% of the metastatic sites (72 of 92). No side effects were observed. RIS was especially useful for detection of lymph node metastases not found by conventional methods and is therefore considered complementary to conventional preoperative staging procedures. PMID- 2552578 TI - [Ptosis--clinical differential diagnosis]. AB - When examining a patient with ptosis the first question to be answered is whether the condition is a simple constitutional variant or a symptom heralding disease. Local diseases of the eyelid may lead to pseudoptosis. The three major disease categories to be considered are: myopathies preferentially affecting the eye muscles; impaired neuromuscular impulse transmission at the motor endplate (myasthenia gravis, botulism); and lesions of the somatomotor (third cranial nerve) and visceromotor (sympathetic) nerve fibers innervating the eyelid muscles. The correct diagnosis may be suspected in the light of a detailed patient history and associated signs on general and neurological examination. Additional investigations may be needed to confirm the diagnosis. PMID- 2552579 TI - [The effectiveness of foot-and-mouth disease vaccines in Switzerland. I. Screening tests and herd immunity]. AB - Protection of the Swiss national cattle herd against foot-and mouth disease is attempted by annual vaccination with inactivated trivalent (O, A, C) vaccines. With the serotype A5 as an example, this paper demonstrates the procedure of potency testing. Serological data obtained with two vaccines in primovaccinated feeder bulls showed that neutralizing antibodies developed within 7 to 14 days post vaccination. There was no statistically significant difference in the anti serotype antibody titers induced by each of the vaccines; differences were seen between anti-O serotype and anti-A serotype antibody titers, regardless of which vaccine was used. Epidemiological analyses with about 3000 cattle demonstrated that single-vaccinated, and to a lesser degree twice-vaccinated, animals often had short lived immunity; that is, the antibody titers waned within a few months. In contrast, the majority of thrice and multiple vaccinated animals maintained relatively high antibody titers throughout the twelve month period of observation. PMID- 2552580 TI - [The effectiveness of foot-and-mouth disease vaccines in Switzerland. II. Stability problems]. AB - One of the vaccines that were used in 1988 to immunize the Swiss national cattle population against foot-and mouth disease (FMD) was apparently not stable. Data, provided by the manufacturer, indicated a high initial antigenic content for serotype O. Protection experiments at the end of the vaccination campaign, however, indicated a substantial loss of serotype O antigen in the vaccine. Serological data, obtained during the campaign indicated that only 12% of the primovaccinated animals and 63% of previously vaccinated animals received an amount of FMD viral antigen sufficient to induce protective immunity. The primovaccinated animals were revaccinated in fall 1988 with a new batch provided by the same manufacturer. The new vaccine induced high titers of neutralizing antibodies in primo- and an anamnestic response in revaccinated cattle. PMID- 2552581 TI - Lectins as cell recognition molecules. AB - Lectins on cell surfaces mediate cell-cell interactions by combining with complementary carbohydrates on apposing cells. They play a key role in the control of various normal and pathological processes in living organisms. PMID- 2552582 TI - Beta-adrenergic receptor kinase: primary structure delineates a multigene family. AB - The beta-adrenergic receptor kinase (beta-ARK), which specifically phosphorylates only the agonist-occupied form of the beta-adrenergic and closely related receptors, appears to be important in mediating rapid agonist-specific (homologous) desensitization. The structure of this enzyme was elucidated by isolating clones from a bovine brain complementary DNA library through the use of oligonucleotide probes derived from partial amino acid sequence. The beta-ARK cDNA codes for a protein of 689 amino acids (79.7 kilodaltons) with a protein kinase catalytic domain that bears greatest sequence similarity to protein kinase C and the cyclic adenosine monophosphate (cyclic AMP)--dependent protein kinase. When this clone was inserted into a mammalian expression vector and transfected into COS-7 cells, a protein that specifically phosphorylated the agonist-occupied form of the beta 2-adrenergic receptor and phosphorylated, much more weakly, the light-bleached form of rhodopsin was expressed. RNA blot analysis revealed a messenger RNA of four kilobases with highest amounts in brain and spleen. Genomic DNA blot analysis also suggests that beta-ARK may be the first sequenced member of a multigene family of receptor kinases. PMID- 2552583 TI - Rare secondary carcinoma from colon to testis. Review of literature and report of a new case. AB - A 65 year old man who underwent a right hemicolectomy for mucus secreting adenocarcinoma of the caecum in 1986 developed a secondary localisation in the right testis one year later. A review of the international literature shows only 14 previously described cases of spread of a colonic tumour into the testis. The Authors discuss the possible mechanisms of this form of dissemination. PMID- 2552584 TI - [Distal radius fracture. Fracture stabilization with biodegradable osteosynthesis pins (Biofix). Experimental studies and initial clinical experiences]. AB - K-wire stabilization and casting of unstable or intraarticular fractures of the distal radius has proven to be an effective method of treatment since many years. However the need of implant removal after fracture healing is a disadvantage of this method. For this reason we investigated on the use of biodegradable implants in a fracture model of the distal radius. The biomechanical testing with Polydioxanon-(PDS)-pins (Ethipin) showed too small primary stability for the use in vivo. Osteosyntheses with three Polyglycolacid-(PGA)-rods Biofix compared with three K-wires 1.6 mm on the other hand showed 82% initial stability. From May to September 1988 typical distal radius fractures in 11 patients have been stabilized with Biofix-rods. The result of primary reduction could be obtained in all cases. These first experiences suggested, that stabilization of distal radius fractures with Biofix-rods might alternatively be performed instead of K-wire pinning. PMID- 2552585 TI - Gestational trophoblastic diseases. PMID- 2552586 TI - Krukenberg tumors: CT features and growth characteristics. AB - We reviewed the computerized tomographic features of pathologically proven metastases to the ovary in 12 patients. Serial CT scans were available in nine of the 12 patients--before removal of the ovaries in five cases (showing typical growth characteristics of these metastases) and afterward in six (showing common patterns of tumor progression). Primary neoplasms metastasizing to the ovary included adenocarcinoma of the colon (seven), stomach (two), appendix (one), and endometrium (one), and carcinoid tumor (one). On CT, metastases to the ovary were large lobulated or oval masses with cystic and solid components. Nine were bilateral and three were unilateral. Three patterns of ovarian enlargement were seen: macrocystic (six), microcystic (three), and predominantly solid enlargement with necrosis (three). Other associated CT findings included carcinomatosis, hydronephrosis, ascites, liver metastases, and lymphadenopathy. The primary tumor in patients without a prior history of malignancy was identifiable retrospectively on CT in four of the five cases. PMID- 2552587 TI - Adenoid cystic carcinoma metastasizing before detection of the primary lesion. AB - Although adenoid cystic carcinomas are occasionally manifested in atypical ways, metastatic disease preceding detection of the primary tumor has not been previously reported. We have described a patient in whom multiple pulmonary metastatic nodules were found one year before identification of primary adenoid cystic carcinoma of the maxilla. This case illustrates the need to include adenoid cystic carcinoma in the differential diagnosis of patients with metastatic disease and an unknown primary lesion. The use of special stains and electron microscopy can be helpful in confirming a diagnosis of adenoid cystic carcinoma. PMID- 2552588 TI - Trousseau's syndrome caused by malignant degeneration of a choledochal cyst. AB - The patient described in this report represents the first reported case of Trousseau's syndrome caused by a malignancy arising in a choledochal cyst. The clinical features were classic, with thrombophlebitis migrans, resistance to anticoagulation, and an occult, mucin-producing adenocarcinoma. The increased recognition of the malignant potential of choledochal cysts should encourage clinicians to consider these unusual anomalies in patients with jaundice or other abnormalities of hepatic function. When possible, the cyst should by excised to minimize the risk of malignant degeneration. PMID- 2552589 TI - [Chemoradiotherapy of disseminated small-cell carcinoma of the lung]. PMID- 2552590 TI - [Quality tumor after-care--dilemma between desire and reality]. PMID- 2552591 TI - Association between human papillomavirus and carcinoma of the oesophagus in South African blacks. A histochemical and immunohistochemical study. AB - Twenty specimens from patients who had undergone oesophagectomy for invasive squamous carcinoma of the oesophagus were examined for morphological evidence of human papillomavirus infection; it was found in 13 specimens. Nineteen specimens showed focal epithelial hyperplasia of the non-neoplastic mucosa. The material was also submitted to immunoperoxidase and modified Feulgen staining to detect viral antigen. Positive Feulgen staining was detected in the superficial layers of the squamous mucosa in 15 specimens, while immunoperoxidase was entirely negative. This demonstrates a possible association between human papillomavirus and oesophageal carcinoma and that the modified Feulgen method may be more sensitive than immunoperoxidase for the detection of viral antigen. Electron microscopy and molecular hybridisation would have to be used for confirmation. PMID- 2552592 TI - Clinical relevance of immunophenotypic and immunogenotypic analysis in acute non lymphoblastic leukaemia. AB - Molecular biology is playing an increasing role in defining pathology today, and has clinical relevance in the routine work-up of patients with malignant diseases, especially those of the blood and lymphatic system. The majority of acute non-lymphoblastic leukaemias (ANLL) express specific myeloid differentiation markers and are terminal deoxynucleotidyl transferase (Tdt) negative. Rarely, some cases are Tdt+ and express the T-cell marker CD7. Although uncommon in Tdt-ANLL, there appears to be a significant incidence of T-cell receptor beta chain (TCR beta) and/or immunoglobulin heavy chain (IgH) gene rearrangements in Tdt+ ANLLs. We have investigated the immunogenotype of 39 patients with ANLL, including 28 from whom immunophenotypic data were available. Thirty-seven of 39 cases had germline IgH and TCR beta genes. Two cases, one with a myeloid/CD7+ CD2+ immunophenotype, had non-germline IgH gene arrangements detectable on Hind III digests only. The possibility of Hind III polymorphisms, or of true somatic gene rearrangement of the IgH gene in these patients, is discussed. Two additional cases had a Tdt+ CD7+ immunophenotype with germline IgH and TCR beta chain genes. Our results confirm the infrequent occurrence of IgH and TCR beta gene rearrangements in ANLL. Expression of Tdt and/or CD7, often in association with IgH gene rearrangements, would appear to identify a subgroup of ANLL patients that respond poorly to standard ANLL therapy and have a poorer prognosis. The diagnostic and prognostic importance of multiparametric analysis in ANLL is emphasised. PMID- 2552593 TI - Fatty acid composition of some South African fresh-water fish. AB - Because of the antithrombotic and anti-inflammatory properties of the n-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), which are found in large quantities in marine fish, the fatty acid composition of the flesh of 18 different species of fresh-water fish found in South Africa was analysed by capillary gas chromatography. In general all the fish studied had low percentages of EPA and DHA and fairly high percentages of arachidonic acid and linoleic acid when compared with some marine fish. The saturated fats constituted 33% of total fatty acids and the mono-enes averaged 35%. The fish studied are therefore not as good a source of n-3 fatty acids as marine fish. PMID- 2552594 TI - CPPD crystal deposition disease. PMID- 2552595 TI - Multiple malignant paragangliomas. A case report. AB - The extra-adrenal paraganglia may be divided into branchiomeric (parasympathetic) and paraxial or para-aortic (sympathetic) networks. A patient is described with two synchronous branchiomeric paragangliomas--a carotid body tumour (glossopharyngeal) and a cardiac chemodectoma (vagus). Both tumours were locally invasive and therefore, by definition, malignant. PMID- 2552596 TI - Latent simian foamy virus. PMID- 2552599 TI - Gastric linitis plastica is not a surgical disease. AB - In a 10-year period, 26 patients with gastric linitis plastica were identified at our institution. The 26 fell into two groups: In group I (nonresection group) seven underwent laparotomy and biopsy, three laparotomy, biopsy, and jejunostomy, and three patients were not explored; in group II (resection group) eight patients underwent total gastrectomy, two subtotal (85%) gastrectomy, one total gastrectomy with thoracic esophagectomy, one total gastrectomy with segmental resection of the tranverse colon, and one total gastrectomy with subtotal pancreatectomy and splenectomy. There was one postoperative death in each group. In group II morbidity consisted of an anastomotic leak in one, pancreatic fistula in two, and pancreatitis in one patient. Mean survival of group I patients was 6.6 months. The mean survival of group II patients was 7.2 months. Patients who underwent total gastrectomy in the presence of peritoneal or liver metastasis lived 4 months. Those who underwent total gastrectomy in the presence of pancreatic involvement lived 4 months. Patients who underwent total gastrectomy for disease limited to the stomach and regional lymph nodes lived 13.6 months. Total gastrectomy for linitis plastica should be performed in those patients who have disease limited to the stomach or regional lymph nodes. Other patients should be offered alternative forms of treatment, including chemotherapy and radiation therapy. PMID- 2552597 TI - [Radiotherapy of glomus jugulare and tympanicum tumors]. AB - Glomus jugulare tumors are difficult to manage therapeutically due to their localisation. Operation may be successful in small tumors but can be hazardous in larger lesions mainly because of bleeding and palsy of cranial nerves. In these cases there should be used radiation therapy under the condition that it is planned by use of computed tomography. Moreover reproducibility of radiation treatment set-up is vital. In this way tumor regression may be achieved. Four own illustrative cases are demonstrated. According to the stage of disease a modified treatment strategy is presented which integrates surgical procedures, angiographic embolization and radiotherapy. PMID- 2552598 TI - Retroviruses and their play-pals. AB - A 28-year-old man with a previous history of Neissena infection presented with diminished vision, disc swelling, and panuveitis. Serologic tests revealed positive titers for both HIV and syphilis. Current epidemiology and treatment of such cases are discussed. PMID- 2552601 TI - Unilateral pulmonary oedema following the removal of a giant pleural tumour. AB - The occurrence of re-expansion pulmonary oedema immediately after excision of a pleural histiocytoma is described. PMID- 2552600 TI - Nedocromil sodium in adults with asthma dependent on inhaled corticosteroids: a double blind, placebo controlled study. AB - Eighty nine adults with asthma who were receiving inhaled corticosteroid and bronchodilator treatment took part in a double blind, randomised, placebo controlled trial of nedocromil sodium, 4 mg four times daily by inhalation. During a run in period of two to four weeks corticosteroid treatment was reduced when possible to produce a comparable level of symptoms across the trial population. The test treatment was then taken for four weeks, with the severity of asthma recorded daily by patients and assessed at two weekly hospital visits. There was an improvement in symptoms in the patients taking nedocromil sodium by comparison with those having the placebo, the differences being significant for diary card PEF readings, asthma symptom scores, and bronchodilator usage at night. The mean difference between the two groups was 18 l/min for PEF, 0.42 for daytime asthma score, and 1.73 puffs in 24 hours for bronchodilator usage. These results suggest that asthmatic patients who require inhaled steroids show better control of their asthma with the addition of nedocromil sodium than of placebo over a four week period after reduction of the dosage of their inhaled steroids. PMID- 2552602 TI - Protein C inhibitor: structure and function. PMID- 2552603 TI - Unfractionated heparin and CY 216: pharmacokinetics and bioavailabilities of the antifactor Xa and IIa effects after intravenous and subcutaneous injection in the rabbit. AB - This report compares the pharmacokinetics and the bioavailabilities of the antifactor Xa and of the antifactor IIa activities generated by intravenous (IV) and subcutaneous (SC) injections of increasing doses of unfractionated heparin (UH) and of a low molecular weight heparin (CY 216). Rabbits were injected with 500, 1,000, 2,500, and 5,000 antifactor Xa u/kg of both heparins and their biological activities were followed at various time intervals. After IV injection the clearance of the antifactor Xa activities was independent of the dose and the clearance of UH was significantly higher than that of CY 216; after SC injection the bioavailability estimated from the antifactor Xa effect was consistently over 100% for CY 216 while that of UH increased from 27% at the lowest dose to 93% at the highest dose. The pharmacokinetic parameters estimated by the antifactor IIa activity of UH were superimposable to those calculated with the antifactor Xa activity. For CY 216 no direct comparison between the two activities was made since the dose injected expressed in antifactor IIa units was 3.4 times lower. UH and CY 216 were therefore injected intravenously to other animals at equivalent and increasing doses expressed in antifactor IIa units (50-5,000 u/kg). The pharmacokinetic parameters calculated from the curves of the antifactor IIa activities were basically identical except at the two lower doses (50 and 100 u/kg) for which UH was cleared faster than CY 216.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552604 TI - Biological activity and safety of the subcutaneous administration of high doses of low molecular weight heparin for 8 days in human volunteers. AB - This study reports on the biological activity and safety of high dose low molecular weight (LMW) heparin therapy administered by two subcutaneous (s.c.) injections daily for 8 days in healthy human volunteers. Group 1 received 2 x 30 aPTT units LMW heparin/kg bodyweight, and group 2 received 2 x 50 aPTT units/kg per day. In group 1, activated partial thromboplastin time (aPTT) and thrombin clotting time (TCT) were uniformly prolonged by 3-5 sec 4 hrs after s.c. administration of heparin. Heptest coagulation time values were prolonged consistently as well by 57 sec on day 1 to 68 sec on day 8. Factor Xa inhibition measured by the S 2222 chromogenic substrate method continuously increased from 0.16 units/ml on day 1 to 0.28 units/ml on day 8. In group 2 prolongation of a aPTT and TCT values increased from 6 sec on day 1 to 15 sec on day 8 and of Heptest time from 70 sec on day 1 to 110 sec on day 8. S 2222 method showed factor Xa inhibitory activity which increased from 0.5 units/ml on day 1 to 0.75 units/ml on day 8. The clinical tolerance of the treatment was good. No changes in clinical chemistry parameters were detected, except for a reversible increase of serum transaminases. The coagulation studies demonstrate accumulation of LMW heparin when high doses are given twice daily. The half life of LMW heparin of factor Xa inhibition increases with increasing doses. Heptest coagulation values were prolonged to 4-6 times the normal values during administration of heparin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552605 TI - Binding of tissue plasminogen activator to cultured human fibroblasts. AB - The binding of 125I-labeled, one-chain tissue plasminogen activator (t-PA) by WI 38 cultured human lung fibroblasts was investigated. Binding of t-PA to WI-38 monolayers was specific, saturable and temperature dependent. One and two-chain t PAs were comparable in their ability to compete with 125I-labeled, one-chain t-PA for binding to fibroblasts, while no inhibition of binding was observed with a 500-fold molar excess of urokinase. Studies with various compounds suggest that neither the catalytic site, the fibrin binding site, nor the carbohydrate moieties on t-PA are involved in its binding to WI-38 cells. At higher temperatures, the amount of cell-bound 125I-t-PA that was removed by either incubation in binding buffer containing an excess of unlabeled t-PA, or by brief treatment with acidic buffer, was small (approximately 20%) suggesting that much of the t-PA is internalized. Electrophoretic analysis of extracts prepared from cells that had been incubated with 125I-t-PA revealed the presence of a major band of 70,000 Mr, which corresponds to intact t-PA. Our results suggest that WI 38 fibroblasts are capable of binding and internalizing t-PA, and that these processes involve a receptor site specific for t-PA. PMID- 2552606 TI - Characterization of N,N'-bis(3-picolyl)-4-methoxy-isophtalamide (picotamide) as a dual thromboxane synthase inhibitor/thromboxane A2 receptor antagonist in human platelets. AB - Picotamide (G137 or N,N'-bis[3-picolyl]-4-methoxy-isophtalamide), a drug which has shown platelet inhibitory effects in vitro and ex vivo, was investigated for its mechanism of action on human platelets in vitro. This compound suppresses the aggregation of human platelets induced by arachidonic acid (IC50: 1.8 x 10(-5) M), low-dose collagen (IC50: 3.5 x 10(-4) M), U46619 (IC50: 1 1.4 x 10(-4) M) and by authentic TxA2 (IC50: 1 x 10(-4) M), without affecting the aggregation induced by A23187 or primary aggregation by ADP. Picotamide inhibits dose-dependently TxA2 synthesis by platelets (IC50: 1.5 x 10(-4) M) and enhances the formation of PGE2. Picotamide-treated platelets also favour the formation of PGI2 by aspirinated endothelial cells; in addition, the drug appears to exert a direct stimulatory effect on PGI2-synthesis, at least at high concentrations. Finally, in platelet-rich plasma stimulated with arachidonic acid, picotamide increases intraplatelet cAMP while no effects on cAMP are detected in unstimulated platelets. In conclusion, picotamide is a dual thromboxane-synthase inhibitor/thromboxane-receptor antagonist in human platelets and introduces a new class of agents potentially useful in antithrombotic therapy. PMID- 2552607 TI - [Severe course of a primary BVD infection in a BVD-seronegative dairy herd]. AB - A severe course of a primary BVD infection in a seronegative dairy herd is reported. Approximately one third of the animals was very ill and showed severe diarrhoea, with temperatures varying from 39 degrees C to 41 degrees C. At the time of the outbreak, twenty-four cows had been pregnant for more than three months. Four animals aborted their calves, two mummies were born and two cows were found not to be pregnant at drying off during the next three months following the outbreak. In addition many of the newborn calves were weak and showed relatively low birth weights. The BVD virus was probably introduced into the herd by a purchased calf, which had pneumonia when it arrived on the farm. PMID- 2552609 TI - [Recurrent hepatitis A]. AB - A 14 year old Turkish girl was diagnosed to have hepatitis A according to symptoms and positive serology. She had a clinical and biochemical relapse eight weeks after the first manifestation of the disease. Relapsing hepatitis A is very rare but it does not alter the usual favourable outcome. PMID- 2552608 TI - [Selected data from childhood epilepsies. ACTH treatment and ketogenic diet: a critical evaluation]. AB - The authors have evaluated critically the results of ACTH and ketogenic diet in 122 cases of 'malignant' childhood epilepsies, especially West-syndrome (WS) and Lennox-Gastaut-syndrome (LGS). In agreement with data from the literature about 10% of the idiopathic forms recovered with or without ACTH. In the other cases ACTH caused a transient amelioration of the EEG and/or the spasms; a high-dosage regimen of ACTH was not significantly better than a low-dosage regimen. Fatal complications only occurred in the high-dosage ACTH group. Ketogenic diet in 10 children with LGS seems to have an influence on the LGS-specific seizures but not on the other signs and symptoms of the syndrome. The diet is difficult to maintain and is not free of side-effects. PMID- 2552610 TI - Two cytotoxic human-human hybridoma antibodies to HLA: TrAH10 (anti A3.1) and TrAG2 (anti B7, Bw42). AB - Two cytotoxic human-human hybridoma IgM antibodies to HLA were generated by EBV transformation of PBMC from multiparous women and fusion of EBV transformed cells with the human fusion partners KR4 or KR12. Both mAbs required the sensitive immunomagnetic cytotoxicity method to display killing of freshly prepared PBMC. One mAb (TrAH10) was specific for HLA-A3. Strikingly, TrAH10 reacted much more strongly with lymphoblastoid cell lines of HLA-A3.1 than of the rare variant HLA A3.2, previously detected by cytotoxic T cells. Thus, in the microcytotoxicity test, the titer of concentrated TrAH10 was approximately 2000 times higher for A3.1 as compared to A3.2, and a clear difference was also observed in radioimmunoassay. Since the two HLA-A3 variants differ by only two amino acids at positions 152 and 156 of the alpha 2-domain's alpha-helix, the epitopes defined by the mAb TrAH10 and HLA-A3.1 specific cytotoxic T cells must be closely related. The observations with TrAH10 suggest that the HLA polymorphism detected by human mAbs may turn out to be as extensive as the T-cell defined HLA polymorphism. The other mAb (TrAG2) bound B7 and Bw42 with equal strength, and in addition bound weakly to some cells that were Bw22 or B39. Magnetic polymerbeads coated with affinity purified human mAbs TrAH10 or TrAG2 formed rosettes with EBV transformed cells carrying relevant HLA antigens; however, rosette formation with freshly isolated PBMC was very weak and unsuitable as a typing assay. PMID- 2552611 TI - A cytotoxic human-human hybridoma antibody (TrC7) specific for HLA-A29. AB - TrC7 is a cytotoxic IgM human-human hybridoma anti-HLA Ab. Its reaction pattern with a panel of 48 HLA-typed EBV-transformed cell lines and PBMC from 348 HLA typed individuals correlated precisely with expression of HLA-A29. TrC7 did not react with HLA-A30, -31, -32 or -w33, which, like A29, are splits of Aw19. PMID- 2552612 TI - Cytomegalovirus infection in low-birth-weight infants with acute respiratory tract disease. AB - To determine a participation of cytomegalovirus (CMV) infection in acute respiratory tract disease (ARTD) of low-birth-weight (LBW) infants, specific antibodies against CMV antigens, IgG antibodies against early antigens of CMV (IgG EA) and IgM antibodies against membrane antigens of CMV (IgG MA) were analyzed. The frequency of IgG EA in patients with ARTD was higher than that in controls (46% vs. 32%), and the geometrical mean titer (GMT) of IgG EA in the patients was also higher than that in controls (50.2 vs. 20.1). Five of 15 ARTD patients had IgM MA, and the frequency was significantly higher than that of controls (33% vs. 1.3%, p less than 0.01). Eleven of 15 LBW patients with ARTD had a history of blood transfusions during the neonatal period, and 5 of them had significant IgM MA indicating active CMV infection. All 4 LBW patients without blood transfusion were negative for IgM MA. These results suggest a close relationship of CMV infection to ARTD of LBW infants, but it remains for further studies whether blood transfusion is a primary source of CMV infection in LBW infants. PMID- 2552613 TI - Blockade of only spinal alpha 1 adrenoceptors is insufficient to attenuate DDT induced alterations in motor function. AB - Male Fischer 344N rats were chronically implanted with an intrathecal cannula and gavaged with p,p'-DDT (1,1,1-trichloro-2,2-bis[p-chlorophenyl]ethane; 30 or 45 mg/kg) or corn oil. Seven hours later, subjects were intrathecally infused with vehicle, 15, 30, 60, or 120 micrograms of prazosin (an alpha 1-adrenergic antagonist). Spectral analysis of bodily movements was performed 7.5, 8, and 10 hr after DDT administration. In control rats, 15 micrograms of prazosin reduced the spectral profiles of spontaneous movements. A 30-micrograms dose produced motor impairments, without significantly changing the spectral profiles. Tremulous movements induced by DDT were unaffected by 15 or 30 micrograms, whereas 60 or 120 micrograms of intrathecal prazosin significantly reduced the spectral profiles of rats pretreated with 45 mg/kg of DDT. Other subjects were administered vehicle or DDT (45 mg/kg), intrathecally infused with 15 or 60 micrograms of prazosin (7 hr), and sacrificed (7.5 hr). Noncannulated rats were gavaged with 60 mg/kg of DDT, injected subcutaneously (sc) with 0.5 mg/kg of prazosin (5.5 hr), and sacrificed (8 hr). Cortical and spinal tissues were used in ex vivo binding assay utilizing [3H]prazosin. Fifteen or sixty micrograms of intrathecal prazosin occupied similar percentages of spinal [3H]prazosin binding sites, but produced a dose-related increase in cortical prazosin equivalents. Sixty micrograms of intrathecal or 0.5 mg/kg of sc prazosin resulted in similar concentrations of cortical prazosin equivalents. Together, these data indicate that while intrathecal prazosin will attenuate DDT-induced motor dysfunction, this effect requires blockade of alpha 1 adrenoceptors in regions other than solely the spinal cord. PMID- 2552614 TI - Specific inhibition of FSH-stimulated cAMP accumulation by delta 9 tetrahydrocannabinol in cultures of rat Sertoli cells. AB - delta 9-Tetrahydrocannabinol (THC), the major psychoactive component in marihuana, is a reproductive toxicant in both man and animals. THC acts at both the level of the pituitary-hypothalamic axis and the testis, specifically the Leydig cell; an effect on the Sertoli cell has not been shown. Since THC inhibits cAMP accumulation in several cell types, we have examined the effect of THC on Sertoli cell function using altered cAMP accumulation as a marker of toxicity. THC reduced the FSH-induced accumulation of cAMP at concentrations which were neither cytotoxic nor affected cellular ATP levels. This inhibition was evident after 3 hr and did not affect the dose of FSH which gave half-maximal stimulation, suggesting that THC does not compete with FSH for binding to its receptor. The ability of THC to inhibit cAMP accumulation was not affected by incubation in the presence of phosphodiesterase inhibitors, making it unlikely that it acts via stimulation of phosphodiesterase activity. This THC-induced inhibition of Sertoli cell cAMP is specific for FSH; it does not affect the ability of forskolin, cholera toxin, isoproterenol, or prostaglandin E1 to stimulate Sertoli cell cAMP. Furthermore, inhibition occurs in the presence of pertussis toxin, suggesting that this effect of THC is independent of the inhibitory adenylate cyclase pathway. Inhibition of Sertoli cell cAMP also occurs with other cannabinoids which are present in marihuana, but which are not psychoactive. These data indicate that a part of the testicular toxicity of THC may be due to a specific alteration of the hormonal control of Sertoli cell function via an inhibition of FSH-stimulated cAMP accumulation. PMID- 2552615 TI - Isolated proximal tubular cells from rat kidney as an in vitro model for studies on nephrotoxicity. I. An improved method for preparation of proximal tubular cells and their functional characterization by alpha-methylglucose uptake. AB - Rat renal proximal tubular cells were isolated by successive EGTA and collagenase perfusions and purified by filtration and isopycnic centrifugation. The method is rapid and provides a much higher fraction of proximal tubular cells (90-95%) than comparable methods. The yield of viable (97 +/- 3%) cells is high (30 X 10(6) cells/g kidney). The intracellular ATP was 16 +/- 2 nmol/mg protein and remained essentially constant for at least 3 hr. The cells were characterized for transport of organic ions and glucose. Glucose transport was studied by alpha [14C]methylglucose uptake; apparent Km and Vmax values were 3.4 +/- 0.5 mM and 4.1 +/- 0.9 nmol/min.mg protein, respectively. This transport could almost be completely inhibited by phloridzin, indicating that the uptake is mediated by the brush border glucose carrier. PMID- 2552617 TI - Bronchoalveolar leukocyte response in experimental silicosis: modulation by a soluble aluminum compound. AB - The biological properties of quartz have been related to its surface reactivity. We have addressed the role of particle surface reactivity in mediating the biological activity of quartz in mixed dusts, by treating the quartz with aluminum lactate. Intratracheal instillation of untreated quartz in rats caused a rapid, sustained alveolitis and bronchoalveolar leukocytes obtained from these animals had enhanced activity in degrading fibronectin, but reduced ability to mount a respiratory burst. Quartz pretreated with aluminum elicited a markedly reduced inflammatory response; the reduced activity of the treated quartz was also reflected in the attenuated change in the key functional parameters, oxidant production and proteolysis of fibronectin. Late intratracheal dosing with aluminum after the quartz-induced alveolitis was well established reduced the inflammatory response and abrogated the effect of quartz on the respiratory burst, but did not alter fibronectin degradation by the leukocytes. Aluminum did not affect the inflammatory response to Corynebacterium parvum and thus the effect was on the quartz particles and not on the inflammatory leukocytes. These findings have implications for the likely pulmonary responses to mixed dusts containing quartz and aluminum silicate clays. PMID- 2552616 TI - Susceptibility to 1,3-butadiene-induced leukemogenesis correlates with endogenous ecotropic retroviral background in the mouse. AB - Previous studies have revealed marked differences in the pattern of carcinogenesis between rats and mice exposed to 1,3-butadiene (BD) that do not appear to be readily explained on the basis of pharmacokinetics or metabolism. Chronic exposure of B6C3F1 mice to BD produces a high incidence of thymic lymphoma (TL) that is not observed in rats. The potential of the endogenous ecotropic retroviral background to influence susceptibility to BD leukemogenesis was examined by comparing the incidence of TL between B6C3F1 and NIH swiss mice. Proviral ecotropic sequences are truncated in the NIH Swiss mouse, and the virus is not expressed. Chronic exposure to BD (1250 ppm) for up to 1 year resulted in a fourfold difference in the incidence of TL between B6C3F1 (57%) and NIH Swiss (14%) mice. These results provide presumptive evidence for retrovirus involvement since NIH Swiss mice lack ecotropic viruses and appear to be relatively resistant to induction of lymphoma by BD. PMID- 2552618 TI - Correlation between the lipophilicity of substituted phenols and their inhibition of the Na+/K+-ATPase of Chinese hamster ovary cells. AB - The Na+/K+-ATPase of Chinese Hamster Ovary (CHO) cells, a plasma membrane bound protein was used as a test system to evaluate the toxicity of several phenol derivatives on membranes. Taking only 2 physico-chemical parameters into consideration, viz., the logarithm of the octanol/water partition coefficient as an indicator for the lipophilicity and the sigma-Hammett constant as a measure for the polarity of the phenol substitutes, it was possible to predict the toxicity with high significance. A multivariate regression analysis calculated a correlation coefficient of 0.99. The results confirm studies performed in our laboratory on cytotoxicity and on functional membrane proteins of fungal and mammalian cells [1,2], suggesting a common mechanism of toxicity by the action of hydrophobic xenobiotics on biomembranes. Taking into account the different sensitivities of the test systems, Quantitative Structure-Activity Relationship (QSAR) analyses could help to explain the basic toxicity of several classes of environmental chemicals. PMID- 2552619 TI - Recognition of D-homoannulation by proton and carbon NMR spectra. AB - One-and two dimensional proton and carbon NMR spectra of the D-homoannulated rearrangement product of triamcinolone (9 alpha-fluoro-11 beta,16 alpha,17 alpha, 21-tetrahydroxy-pregna-1,4-diene-3,20-dione) establish its structure as that of 9 alpha-fluoro-11 beta,16 alpha,17 alpha-trihydroxy-17 beta-hydroxy-methyl-D homoandrosta-1,4-diene-3,17 alpha-dione. These methods accord ready recognition of D-homoannulation of C21-17-hydroxy-20-ketosteroids. PMID- 2552620 TI - Membrane fluidity alterations in a cytochrome P-450-deficient mutant of Candida albicans. AB - A cytochrome P450-deficient mutant of the pathogenic fungus, Candida albicans, which accumulates exclusively 14 alpha-methylsterols in place of the normal end product sterol, ergosterol, was examined for alterations in membrane fluidity by electron paramagnetic resonance. The results using four nitroxyl spin labels indicated that exponential phase cultures of the mutant strain, D10, had a uniformly more rigid membrane than similarly grown wild type. Since D10 shows a sterol spectrum similar to that of wild type cells treated with imidazole and triazole antifungal agents, many of the physiological effects reported as the result of azole application may be the result of alterations in membrane fluidity. PMID- 2552621 TI - Human leukemia and retroviruses. PMID- 2552622 TI - Cell-activation responses to cytomegalovirus infection relationship to the phasing of CMV replication and to the induction of cellular damage. PMID- 2552623 TI - Association of bluetongue virus with the cytoskeleton. PMID- 2552624 TI - Morphogenesis of flaviviruses. PMID- 2552625 TI - Cytomegalovirus infections in seropositive patients after transfusion. The effect of red cell storage and volume. AB - Antibody responses to cytomegalovirus (CMV) after red cell (RBC) transfusion were studied in 84 seropositive surgery patients and 82 seropositive oncology patients. The surgery patients were randomized to receive RBCs stored either 3 to 8 or 20 to 42 days after donation. Of 38 patients receiving RBCs stored 8 days or less, 3 developed a rise in titer (4-fold increase) of IgG antibody to CMV 8 to 12 weeks after transfusion. This rate of response (8%) did not differ significantly (p = 0.23) from that (16%) in the 46 patients receiving RBCs stored 20 to 42 days. Seropositive oncology patients were randomized to receive RBCs from seronegative or random donors. Five (19%) of 27 oncology patients receiving seronegative RBCs and 13 (23%) of 55 patients receiving random RBCs (mean, 2 seropositive RBC units/patient) developed a rise in titer of antibody to CMV. No CMV morbidity occurred in either patient group. For both patient groups, a rise in titer of antibody to CMV was associated with the number of transfused RBC units. These results confirm that CMV-seronegative RBCs are unnecessary for infrequently transfused seropositive patients. They also suggest that multiple transfusions of stored RBCs are as immunosuppressive as multiple transfusions of RBCs used within a few days after donation. PMID- 2552626 TI - Alternative splicing or cloning artefact? PMID- 2552627 TI - Neopterin excretion after liver transplantation and its value in differential diagnosis of complications. AB - The aim of this study was to investigate patterns and clinical utility of a neopterin marker in liver allograft recipients. Urinary neopterin levels were studied in 59 transplant patients daily until discharge. During the early postoperative period (days +1 to +20) neopterin excretion exhibited a liver specific pattern that clearly differed from that seen in renal and bone marrow transplant recipients. Neopterin concentrations increased and reached peak values on day +7. The height of this early peak varied in context with complications such as infection or rejection, and was correlated with an unfavorable prognosis. In contrast to the early phase, no liver-specific pattern was observed after day 20. As seen in all other transplant patient populations, values normalized in patients with an uncomplicated course but again increased during infection or rejection episodes. Neopterin levels were particularly high during CMV infection, and their increments were directly related to the severity of CMV disease. The neopterin marker, although not specific, enables discrimination between patients with a complicated and uncomplicated clinical course. High neopterin values early after transplantation are associated with unfavorable prognosis and correlate with an increased risk of developing CMV disease. PMID- 2552628 TI - Successful treatment of invasive fungal infection with fluconazole in organ transplant recipients. PMID- 2552629 TI - Development of foot-and-mouth disease virus strain characterisation--a review. PMID- 2552630 TI - Prevalence of antibodies in goats in north-eastern Brazil to selected viral and bacterial agents. PMID- 2552631 TI - The unusual spectrum of neuroendocrine lung neoplasms. AB - Neoplasms of the lungs showing neuroendocrine differentiation are classified histologically into the following groups: (1) carcinoid, (2) atypical carcinoid (well-differentiated neuroendocrine carcinoma and malignant carcinoid, (3) small cell neuroendocrine carcinoma (small cell undifferentiated carcinoma and oat cell carcinoma), and (4) large cell neuroendocrine carcinoma (atypical endocrine tumor of the lung and intermediate neuroendocrine carcinoma). Nine examples of neuroendocrine lung carcinomas are discussed that have unusual histologic features that make it difficult to assign them to one of the above groups, have unusual immunohistochemical features, have unusual ultrastructural features, or exhibit a biologic behavior different from what one would have predicted from their morphologic appearance. The findings in these nine cases suggest that the present classification of neuroendocrine lung neoplasms may be too precise and that these neoplasms, like other nonneuroendocrine pulmonary tumors, exhibit a wider morphologic and biologic spectrum than previously appreciated. PMID- 2552632 TI - Myofibroblastoma of the meninges. PMID- 2552633 TI - Mechanism of tolerance to renal ischemia in harbor seal: role of membranes. AB - Many diving animals are capable of engaging in long breath-hold dives. Investigations were undertaken to compare the active tubular transport of organic ions between kidney slices from harbor seals (Phoca vitulina) and rats (control) under anoxia or low pH or both. Para-aminohippurate (PAH) and tetraethylammonium (TEA) were used as a representative organic anion and cation, respectively. The slice-to-medium concentration ratios (S:M) of PAH (congruent to 2.5) in the harbor seal was not altered (unlike in the rat) after 60 min of incubation in a nitrogen atmosphere but was reduced to congruent to 1.0 in the presence of 10(-4) M 2,4-dinitrophenol. Subsequent studies also demonstrated that PAH uptake in the harbor seal was not influenced by pH of the incubation medium over a range of 6.0 to 7.4. In contrast, PAH uptake was very sensitive to the medium pH in the rat, decreasing significantly with reduction in pH. Although the TEA uptake decreased in anoxia in both species, it decreased considerably less in the harbor seal than in the rat (40 vs. 70%). However, TEA uptake decreased significantly at low pH in both species. The magnitudes of increase in intracellular Na and of decrease in intracellular K in anoxia were consistently less in the harbor seal than in the rat, especially at low pH. On the basis of these and other findings reported in the literature, cellular mechanisms for the remarkable ischemic tolerance of the seal kidney are discussed. PMID- 2552634 TI - Effects of the pH and the urine infected by Escherichia coli and Proteus mirabilis on chromic catgut, polyglycolic acid and polyglactin 910: study in vitro. AB - In order to study the effects of the pH and the urine infected by Escherichia coli and Proteus mirabilis on chromic catgut, polyglycolic acid (PGA) and polyglactin 910 (P910), we divided the experiment into three steps. In the first step, the behavior of suture material immersed in sterile urine, urine infected by E. coli and urine infected by P. mirabilis and in culture environment infected by P. mirabilis was studied. The physical features were observed continuously up to the 6th day. In the second step, every element of the urea-splitting reaction was isolatedly studied , without the presence of bacterial agents. And in the last step, that reaction was mimetized in sterile environments and in environments with acid and alkaline pH. While the chromic catgut was kept integral in all the environments, the PGA and the P910 dissolved in urine infected by Proteus, which was caused by the ammonia resulting from the urea splitting reaction. This dissolution was also observed in sterile environment (mimetization of the urea-splitting reaction by urease, with no Proteus). The destruction of the sutures was not influenced by the pH variance. PMID- 2552635 TI - [The spontaneous course of bronchial cancer. A study of survival time]. AB - In order to assess the success of therapy in cases of bronchial carcinoma, the clinical course of 38 untreated patients was studied retrospectively. We were unable to demonstrate a significant difference in the survival rate between patients who underwent radical therapy and those who were not treated at all. PMID- 2552636 TI - [Artificial synthetic bone tissue]. PMID- 2552637 TI - [Cell-mediated immunity in calves immunized against or infected with the bovine rhinotracheitis virus]. AB - Simple and expeditious methods--leucocyte adherence inhibition LAI and leucocyte migration inhibition LMI--are described in the present paper; these methods enable to investigate cell-mediated immunity of animals. The two tests, along with serological and virological methods, were used to study changes in the immunity of calves after experimental infection by IBRV and after immunization by inactivated oil vaccine against IBRV. The results have indicated that the changes in cell-mediated immunity after experimental infection and vaccination of calves assumed courses independent of the changes in humoral reaction. Total cell mediated immunity reaction started earlier in experimentally infected calves than in vaccinated ones. In some vaccinated calves a modified course of cellular immunity reactions was observed, characterized also by elimination of IBR virus on the nasal mucous membrane after challenge infection. The LAI and LMI test can be recommended for immunological monitoring in clinical laboratories and in research. PMID- 2552638 TI - [The effect of natural zeolite on the excretion and distribution of radiocesium in rats]. AB - Observation was made of the influence of natural zeolite (clinoptilolite) supplement in food on 134Cs excretion and distribution after oral internal contamination of laboratory brown rats. After diet administration with 2.5, 5, and 10% zeolite supplement the 134Cs elimination in droppings increased and the radionuclide deposition in liver, kidneys and femoral musculature decreased. The zeolite decontamination effects were observed with preventive administration, as well as with sorbent administration from the 24th hour after a single internal contamination. PMID- 2552640 TI - Outbreak of virulent infectious bursal disease in East Anglia. PMID- 2552639 TI - Development of a neutralising antibody response to an inoculated cytopathic strain of bovine virus diarrhoea virus. AB - Fourteen clinically healthy cattle that were persistently infected with non cytopathic bovine virus diarrhoea virus (BVDV) and three BVDV-free cattle were inoculated with one of three cytopathic BVDV strains. Mucosal disease developed in 12 of the viraemic cattle, resulting in a moribund condition 17 to 99 days after inoculation. Two of the viraemic cattle remained clinically healthy until the end of the experiment, 14 months after inoculation. The BVDV-free cattle did not develop clinical signs after inoculation. From each cow with mucosal disease a noncytopathic and a cytopathic BVDV strain were isolated from tissue specimens collected post mortem. All the cattle developed moderate to high levels of neutralising antibodies against the cytopathic BVDV strain with which they were inoculated. The antibodies from 10 of the 12 cattle with mucosal disease did not react with the cytopathic BVDV strains isolated post mortem, and antibodies from none of them reacted with the non-cytopathic BVDV isolates. Antibody responses to the inoculated BVDV strains developed earlier in the viraemic cattle than in the BVDV-free cattle. PMID- 2552641 TI - Acute infectious bursal disease in poultry. PMID- 2552642 TI - Misleading labelling. PMID- 2552643 TI - Vaccination and eradication programme against Aujeszky's disease in Sweden, based on a gI ELISA test. AB - A killed gI-negative vaccine combined with a gI enzyme-linked immunosorbent assay (ELISA) test was used for the first time in Sweden in an attempt to eradicate Aujeszky's disease from a weaner pig producing herd. The herd had experienced three severe outbreaks of the disease during a 10 year period and at the start of the programme 96 per cent of the herd's 104 breeding animals were seropositive to the Aujeszky's virus. In addition, there was serological evidence of active virus circulation among younger animals. During the programme, all breeding animals were vaccinated every sixth month and replacement animals were tested free of disease and vaccinated before entry into the herd. When the originally seropositive animals had been rotated out of the herd, all breeding animals and a sample of weaner pigs were tested twice at six weeks' interval. No seroconversions to gI had taken place and the herd was declared Aujeszky's disease-free, 22 months after the start of the programme. PMID- 2552644 TI - Nephroblastoma in a calf. PMID- 2552645 TI - Characterization of porcine and some ruminant pestiviruses by cross neutralization. AB - Serologic relationships between 11 pestivirus strains that originated from pigs and five that originated from cattle or sheep were studied by cross neutralization. Experiments were performed with pig and sheep sera raised against the strains. The results were analysed by a computerized taxonomic procedure. The 16 viruses were classified into four distinct serologic groups. All hog cholera virus (HCV) strains were classified in one group; the other three groups consisted of strains that can infect pigs, but that are identified as bovine viral diarrhoea virus (BVDV) or border disease virus (BDV), or showed a closer relationship to BVDV and BDV than to HCV. PMID- 2552646 TI - Investigations of bluetongue and other arboviruses in the blood and semen of naturally infected bulls. AB - Small groups of bulls were exposed to natural infection with arboviruses. The bulls were bled and ejaculated regularly and the blood and semen were processed for virus isolation. Over a 5-year observation period, virus isolation and serology indicated that the 29 exposed bulls had experienced 79 viraemic episodes with the viruses of the bluetongue, epizootic haemorrhagic disease, Palyam and Simbu serogroups and an incompletely characterised rhabdovirus. In no instance was there unequivocal evidence of bluetongue virus contamination of semen, despite 18 infections in the study period. PMID- 2552647 TI - Oligonucleotide fingerprint analysis of the genomes of two variants of bluetongue virus serotype 2 isolated in the U.S.A. AB - The genome segments of two electrophoretically distinct variants of bluetongue virus (BTV) Serotype 2 (Ona A and Ona B) from the U.S.A. were analyzed by double dimension gel electrophoresis of RNase T1 produced oligonucleotides. Segments 1, 4, 5, 6, 7 and 10 were examined individually after separation by SDS-PAGE; and Segments 2 and 3, and 8 and 9, which were difficult to resolve, were fingerprinted as pairs. The Ona A and Ona B strains appeared to be closely related since corresponding segments were comparable, sharing 53-89% of the large oligonucleotides counted. Since the strains with the Ona A electropherotype preceded Ona B infection in Florida, U.S.A. and since Ona A was indistinguishable from the early African isolate of Serotype 2, Ona B was thought to be a variant of an Ona A strain. These data tend to support the hypothesis that Ona B could have evolved from Ona A as the result of point mutations or genetic drift. PMID- 2552648 TI - [Roentgenologic characteristics of small bronchoalveolar cancer of the lung under conditions of image processing using linear filtration (roentgenomorphologic comparison)]. AB - X-ray and morphological signs were compared in 50 patients with histologically verified bronchioloalveolar cancer (BAC). Particular attention was drawn to a study of an x-ray picture after computer processing of a tomographic image using a special technique. The most characteristic x-ray signs of BAC were subpleural localization, the presence of a fragmented scar in a tumor, and ill defined outlines. The peculiarities of an x-ray picture of small-size BAC were determined by the morphological structure of a tumor, a degree of its differentiation that could be detected in most cases only after image processing by the linear filtration technique. PMID- 2552649 TI - [Bronchoalveolar cancer (clinical, x-ray and morphologic characterization)]. AB - The authors presented a clinicoroentgenological picture of bronchioloalveolar cancer based on the analysis of 154 cases; 4 types of disease were defined: peripheral tumors, pseudopneumonic type tumors, multiple nodular formations, and mixed type tumors. Common features of this disease and symptomatology of each separate type were described. The most prominent features of bronchioloalveolar cancer are the absence of bronchial tree involvement and frequent presence of typical cavities with well defined outlines and "lattice" structure. The characteristic features of peripheral bronchioloalveolar cancer are slow growth rates, subpleural localization, inhomogeneous structure, and irregular outlines. The mixed type (far advanced tumors) is characterized by the combination of lobular, spheroidal and microfocal changes. Early recognition of definite types of bronchioloalveolar cancer ensures timely surgical treatment and prevents disease progression to disseminated bilateral lesions. PMID- 2552650 TI - Evaluation of immunity to feline infectious peritonitis in cats with cutaneous viral-induced delayed hypersensitivity. AB - Delayed-type hypersensitivity (DTH)-like reactions to feline infectious peritonitis (FIP) virus (FIPV) were induced in the skin of nine cats that were asymptomatic after a previous challenge-exposure with FIPV. Four of the nine previously challenge-exposed cats were negative for virus-neutralizing antibodies against FIPV at the time of intradermal (ID) testing for DTH. Two other cats tested for DTH when acutely ill with clinical FIP did not have cutaneous DTH responses to FIPV. Gross skin reactions to FIPV injected ID were observed in six of nine asymptomatic cats (67%) at postintradermal inoculation hours (PIH) 24, 48, and/or 72. The reactions consisted of focal, 1-5-mm to 2.5-cm diameter indurated or semi-firm, nonerythematous, slightly raised nodules. Microscopically, DTH-like reactions were observed in biopsies taken from the FIPV inoculated skin of asymptomatic cats at PIH 24 to 72. The lesions consisted of perivascular and diffuse dermal infiltrations by macrophages, lymphocytes, and polymorphonuclear leukocytes (PMN). The dermal infiltrates, which were maximal at PIH 48 or 72, were predominantly mixed inflammatory cells (five of nine cats) or PMN (four of nine cats) at PIH 24, but later were predominantly mononuclear cells (six of nine cats) or mixed inflammatory cells (two of nine cats) at PIH 72. Five of nine cats (56%) with positive DTH skin responses had increased survival times after lethal ID challenge-exposure with FIPV compared to mean survival times in FIPV-naive, non-immune control cats that were DTH-negative when ID challenge exposed. Four of nine DTH-positive cats (44%) resisted an ID challenge-exposure dose of FIPV that was fatal in both control cats, and two of the four remaining DTH-positive cats survived a third challenge-exposure with highly lethal doses of FIPV given intraperitoneally. Four of the six DTH-positive cats (67%) that died after re-challenge and were necropsied had lesions of noneffusive FIP, suggesting that cellular immunity may also be involved in the pathogenesis of noneffusive disease, whereas both control cats and both DTH-negative cats with clinical disease succumbed to effusive FIP. Seemingly, DTH responses to FIPV can be associated with an increased level of resistance to disease; however, this state of immunity is variable and apparently can be lost with time in some cats. PMID- 2552651 TI - Cell mediated immune responses in ponies following infection with equine influenza virus (H3N8): the influence of induction culture conditions on the properties of cytotoxic effector cells. AB - Cytotoxic cell precursors and/or cytotoxic memory cells were demonstrated in the peripheral blood of ponies after aerosol infection with influenza A/equine/Newmarket/79 (H3N8). In order to reveal their cytotoxic potential, peripheral blood mononuclear cells required a secondary antigenic stimulation. In vitro induced cytotoxic cells showed activity against influenza infected target cells in a 3-4 h 51Cr-release assay. The reactivity of cytotoxic cells was markedly influenced by the conditions of the secondary induction culture. If high concentrations of exogenous crude equine IL-2 were used, virus infected target cells were susceptible to lysis by autologous or allogeneic effector cells. However, if IL-2 concentration was reduced, cytotoxic cells were generated which showed features consistent with cytotoxic T cells in that target-cell killing was genetically restricted. PMID- 2552653 TI - Ultrastructural observations on luminal structures of pleomorphic adenoma of parotid and submandibular salivary glands of man. AB - Luminal structures found in salivary pleomorphic adenomas consisted of lumina surrounded by epithelial cells that varied from being packed together to being widely separated except at the luminal margin. Communication between lumina and the surrounding stroma was occasionally seen. Secretory material and cellular debris were seen in lumina, invaginations of the luminal surfaces of periluminal cells, associated vesicles, and vacuoles. Secretory granules, lysosomes and lipofuscin were seen in periluminal cells. Secretory material and debris from necrotic periluminal cells appear to accumulate in lumina, and to be endocytosed and degraded lysosomally by periluminal cells. The finding of communications between lumina and the surrounding stroma suggests that the stromalization of the epithelium includes the luminal structures. The present investigation supports the hypothesis that many of the cellular features of the pleomorphic adenoma relate to the microenvironment. PMID- 2552652 TI - Nucleolar grading of breast cancer. Comparative studies on frequency and localization of nucleoli and histology, stage, hormonal receptor status and lectin histochemistry. AB - The localization and number of nucleoli of tumour nuclei were examined after conventional histological typing of breast carcinomas (WHO classification), staging, biochemical receptor status, and lectin binding site histochemistry. With increasing histological atypia and grade of malignancy, the frequency of nucleoli increased significantly and their location shifted from the center to the nuclear periphery. These changes correlated with an increase in indices of mitosis and cell loss. The nucleolar size correlated with the increase in the grade of malignancy. Hormone receptor and lectin binding site positive carcinomas were characterized by a low nucleolar frequency and by small nucleoli in central position, whereas carcinomas with high grades of malignancy and negative hormone receptor and lectin binding site status showed large, often multiple nucleoli in eccentrical position at a high frequency. The studies have demonstrated that nucleolar status is an easily practicable histological/cytological method for use in the assessment of prognosis of carcinomas of the breast. PMID- 2552654 TI - Genetic and phenotypic studies on Ross River virus variants of enhanced virulence selected during mouse passage. AB - We have passaged Ross River virus (RRV) in mice to generate variants with increased mouse virulence and attempted to relate changes in virulence to genome sequence changes. RRV NBO (zero passage in mice) is a plaque-purified clone of the mouse-avirulent strain RRV NB5092, and is of low virulence for day-old mice. During RRV NBO replication in infant mice, its virulence for day-old mice increased markedly with time. By 7 days postinfection the LD50 value of harvested virus (passage level one) was congruent to 10(4)-fold less than that of the parental virus. No further decrease in LD50 followed 10 serial passages in infant mice. However, 10th passage level virus showed increased clinical effects in week old mice by comparison with virus from passage levels one and two. The growth kinetics of RRV variants in mice suggested that the rate and extent of RRV replication in the brain tissue determined the enhanced mouse virulence of serially passaged virus. Seven out of eight independently passaged, 10th passage level variants had changes in the E2 gene leading to one or two amino acid substitutions. The changes were at residues 212, 232, 234, 251, 341, 27 and 172, and 72 and 134 in these variants; all changes except two were nonconservative. Residues 212, 234, and 251 form part of a neutralization determinant in RRV. Changes in epitope b2 (which includes amino acids 246, 248, and 251) alter the kinetics of RRV entry into cells (P. Kerr, R. C. Weir, and L. Dalgarno, unpublished data). First and second passage level virus of enhanced virulence was unchanged in E2 or E1 gene sequences from RRV NBO. However, 1st, 2nd, and 10th passage level virus induced higher levels of virus-specific RNA synthesis than did RRV NBO in cultured BHK cells. We propose a model for the mechanism of virulence enhancement on passaging RRV NBO in mice. PMID- 2552655 TI - Reversion of the attenuated and temperature-sensitive phenotypes of the Sabin type 3 strain of poliovirus in vaccinees. AB - Isolates of type 3 poliovirus from vaccine-recipients were characterized in terms of virulence, sensitivity of growth to high temperatures, and differences in genome structure from the Sabin type 3 vaccine strain. These included point mutations in the region of the genome coding for the structural proteins and in the 5' noncoding region, and the presence of type 1 or type 2 poliovirus genomic sequences resulting from intertypic recombination. Isolates from healthy vaccinees resembled those from vaccine-associated cases of poliomyelitis in all of these properties. Suppression of the temperature-sensitive phenotype was strictly correlated with reversion to virulence in nonrecombinant type 3 strains. Recombinant isolates were more attenuated than expected, even when they had lost all mutations known to attenuate the type 3 vaccine strain. PMID- 2552656 TI - Identification of the major genetic determinant for neurovirulence of tick-borne orbiviruses. AB - Three members of the Great Island antigenic subgroup (Kemerovo serogroup) of tick borne obiviruses produced fatal infections following intracerebral inculation of 2-day-old mice. The average survival times and PFU/LD50 ratios of mice inoculated with Wexford (WEX) virus were significantly greater than those of either Nugget (NUG) or Great Island (GI) virus. Reassortant viruses were isolated following dual infections of cell cultures with a spontaneous temperature-sensitive (ts) mutant of WEX virus, and either NUG wild-type virus or a ts mutant of GI virus. The neurovirulence for mice and derivation of the genomic RNA segments were determined for the reassortants. Analysis of this data revealed that the pathogenic phenotype of the reassortant viruses depended on the parental origin of genomic segment 4. The major genetic determinant of serotype specificity was not shown to influence neurovirulence in these investigations. PMID- 2552657 TI - Herpes simplex virus neurovirulence and productive infection of neural cells is associated with a function which maps between 0.82 and 0.832 map units on the HSV genome. AB - A herpes simplex virus (HSV) intertypic recombinant (RE6) has been shown to be completely and specifically non-neurovirulent in mice. Direct intracranial inoculation of 10(8) PFU of RE6 does not result in a lethal encephalitis. Neurovirulent recombinant viruses were generated by cotransfection of RE6 DNA with DNA fragments cloned from the pathogenic HSV-1 strain 17 syn+. It was found that a 1.6-kb fragment mapping between 0.82-0.832 m.u. could restore the neurovirulent phenotype. Recombinants which incorporated at least part of this fragment were at least 100,000-fold more neurovirulent than RE6. The recombinants displayed a greatly enhanced capacity to replicate in mouse brain in vivo, but did not display enhanced replication over that of RE6 in cultured mouse cells at 38.5 degrees. Immunohistochemical analysis of infected mouse brain tissue revealed that the permissive host cell range of the recombinants was dramatically altered from that of RE6. While antigen positive cells were extremely rare in mouse brain tissue infected with RE6, the neurovirulent recombinants produced antigens in many cell types including neurons. Thus, wild-type HSV-1 sequences mapping between 0.82-0.832 m.u. can donate a highly neurovirulent phenotype to RE6. PMID- 2552658 TI - Characterization of cDNAs of spliced HPV-11 E2 mRNA and other HPV mRNAs recovered via retrovirus-mediated gene transfer. AB - Human papillomaviruses (HPVs) are associated with hyperproliferations of cutaneous or mucosal epithelium. These viruses cannot be propagated in any cell culture system. Because cloning cDNA copies of HPV mRNAs recovered from human lesions has met with only very limited success, the characterization of HPV mRNAs has been problematic. Using the Moloney murine leukemia virus vector system (C.L. Cepko, B.E. Roberts, and R.C. Mulligan, 1984, Cell 37, 1053-1062), we have recovered cDNAs of spliced E2 mRNAs of human papillomavirus type 11 and additional mRNAs of type 11 and type 18 and determined the utilization of open reading frames (ORFs) in the DNA sequences. The recovery of cDNA copies of messages with splice sites identical to those previously described strongly suggests that the newly characterized splice donors and acceptors are also authentic. The HPV-11 E2 cDNA contains the intact E6 and E7 ORFs and the beginning of the E1 ORF in the first exon, which is then spliced from nt 847 to the second exon at nt 2622, 100 nucleotides upstream from the initiation codon for the E2 ORF. The initiation codon in the E1 ORF is followed by four additional in-frame AUG triplets and an in-frame termination codon positioned 30 nucleotides upstream from the initiation codon for the E2 protein. The authenticity of this putative E2 cDNA was shown by its ability to provide enhancer transactivating activity in chloramphenicol acetyltransferase (CAT) assays in several cell lines. A mutation in the genomic DNA at this splice acceptor site eliminates its activity, demonstrating that the splice is essential for the expression of the E2 protein. We conclude that the translation of the HPV-11 E2 protein requires internal initiation. PMID- 2552659 TI - Characterization of rare human papillomavirus type 11 mRNAs coding for regulatory and structural proteins, using the polymerase chain reaction. AB - Certain human papillomavirus (HPV) types cause warts, dysplasias, and carcinomas of the ano-genital and oral mucosa. Because of the inability to propagate HPVs in cultured cells, the paucity of viral mRNAs in human lesions, and the complexity of alternatively spliced transcripts derived from different promoters, it has not been possible to ascertain the exact structures of the majority of the mRNA species and the proteins encoded. We have adapted the recently developed polymerase chain reaction to amplify cDNAs of rare, type 11 HPV mRNAs isolated from a productively infected human foreskin xenograft in an athymic mouse. The oligonucleotide primers were designed to flank each of the mRNA splice sites previously mapped by electron microscopic analysis of heteroduplexes formed between cloned HPV-11 DNA and viral mRNAs isolated from genital warts. The splice junctions were determined by direct sequencing of the PCR-amplified cDNA products or after the cDNA was cloned into a plasmid vector. We provide the first direct evidence for the existence of rare mRNAs with the potential to encode regulatory proteins that have been hypothesized to exist for HPVs. Depending on the lengths of the upstream exons, the translation frame used and the possibility of internal reinitiation during translation, one pair of mRNAs with the same splice junction could encode the viral DNA copy number modulating protein E1-M, the enhancer repression protein E2-C, or both. A second pair of mRNAs, also with identical splice junctions, encode the enhancer-regulating protein E2; the longer of the two could also encode, in its 5' exon, either or both of the E6 and E7 proteins. Finally, we demonstrate that the doubly spliced late message for the major virion capsid protein L1 also contains the entire coding region for the early E1 E4 protein in the first two exons, with the initiation codon for the L1 protein located precisely at the splice acceptor of the third exon. The potential of this late mRNA to encode both the E1 E4 protein and the capsid protein could contribute to the preponderance of the E4 protein in the lesion. PMID- 2552660 TI - Vaccinia virus gene D12L encodes the small subunit of the viral mRNA capping enzyme. AB - Vaccinia virus gene D12L, which lies between nucleotides 14,350 and 13,487 in the HindIII D fragment, is transcribed at early times in infection and is capable of encoding a protein 287 amino acids in length with a predicted molecular mass of 33,331. A polyclonal antiserum was raised in rabbits to a fusion protein containing 279 amino acids of the D12L protein, and this serum was used to investigate both the time of synthesis and the function of the D12L protein. A combination of Western blot analysis and immunoprecipitation from pulse-labeled and pulse-chased cell extracts demonstrated that the synthesis of a 31-kDa protein begins early in infection, that it reaches a plateau by about 4 hr, and that it is stable in the infected cell. The D12L protein was localized by Western blot analysis of detergent-solubilized virions to the sodium deoxycholate soluble fraction which suggested that it may be a virion core-associated enzyme. Due to the similarity in apparent molecular weight between the D12L protein and the small subunit of the vaccinia mRNA capping complex the anti-D12L antiserum was employed in Western blot analysis of fractions generated during the purification of the virion mRNA capping enzyme. The 31-kDa D12L protein copurified with the virus capping enzyme through chromatography on heparin-agarose and phosphocellulose and also cosedimented with the capping enzyme through a glycerol density gradient. In addition, the anti-D12L antiserum coprecipitated the large subunit of the capping enzyme, confirming that gene D12L encodes the small subunit of the viral mRNA capping enzyme. An insertion mutation which destroys the gene D12L coding sequence was constructed in a plasmid containing a portion of both genes D11L and D12L and this plasmid was used to rescue a ts mutation, in a single step, in the adjacent gene D11L. Southern blot analysis of the re-plaque purified virus permitted the identification of the mutant virus only when the mutant was propagated in the presence of wild-type helper virus. We concluded from these data that gene D12L is essential for virus propagation in tissue culture. PMID- 2552661 TI - Localization of conserved and variable antigenic domains of equine infectious anemia virus envelope glycoproteins using recombinant env-encoded protein fragments produced in Escherichia coli. AB - Previous characterizations of equine infectious anemia virus (EIAV) glycoprotein variation by DNA sequence analysis and epitope mapping using monoclonal antibodies (MAbs) have revealed the presence of conserved and variable regions within the EIAV env gene. To extend these studies, fragments of the EIAV envelope proteins gp90 and gp45 were expressed in Escherichia coli and used in Western blot analysis with a diverse panel of equine immune sera to identify antigenic segments. All sera from EIAV-infected animals reacted with the carboxyl terminal portion of gp90 and the amino terminal portion of gp45, indicating the highly conserved and immunodominant nature of these regions. Other gp90 segments, both from conserved and variable env sequences, displayed variable reactivities with the panel of equine sera. A panel of MAbs was also used in Western blot assays with the recombinant protein fragments for physical localization of previously identified MAb epitopes. The binding sites of two neutralizing MAbs were localized to a highly variable region of gp90, while nonneutralizing epitopes were localized to conserved and variable regions of the envelope glycoproteins. These results, in addition to localizing important antigenic sites on EIAV glycoproteins, indicate that previously defined conserved and variable env nucleotide sequences indeed encode protein sequences constituting conserved and variable immunogens during persistent infection by EIAV. PMID- 2552663 TI - The 3'terminus of lactate dehydrogenase-elevating virus genome RNA does not contain togavirus or flavivirus conserved sequences. AB - Lactate dehydrogenase-elevating virus (LDV) is currently considered to be an unclassified togavirus. The 3' terminus of the genome RNA of the C-strain of LDV was cloned and sequenced. A synthetic DNA oligomer complementary to the 3' portion of this cloned sequence was then used to prime dideoxy sequencing from the LDV-C genome RNA as well as from the genome RNAs of three additional LDV isolates. A high degree of sequence conservation was observed in the 3' terminal region among the four LDV isolates analyzed. Comparison of the LDV 3' sequence with those of the alpha togaviruses, rubella virus, and the flaviviruses showed that the LDV genome does not contain conserved 3' sequences characteristic of these viruses. PMID- 2552662 TI - Characterization of rotavirus replication intermediates: a model for the assembly of single-shelled particles. AB - The segmented double-stranded (ds)RNA genome of the rotaviruses is replicated asymmetrically with viral mRNA serving as the template for minus-strand RNA synthesis. To identify intermediate structures in rotavirus replication, subviral particles (SVPs) purified from the cytoplasm of simian rotavirus SA11-infected cells were assayed for RNA polymerase activity in a cell-free system that supports viral RNA replication. Intact SVPs containing newly made RNA were resolved by electrophoresis under nondenaturing conditions on 0.6% agarose gels (50 mM Tris-glycine, pH 8.8). This gel system was found to separate without disrupting SA11 single- and double-shelled virions and virion-derived core particles. SVPs from the cell-free system that contained newly made dsRNA migrated in the agarose gels at positions between virion-derived cores and intermediate of single- and double-shelled virions. SVPs containing newly made dsRNA were eluted from the gel and analyzed for protein content by electrophoresis on polyacrylamide gels. The results showed that three distinct types of replication intermediates (RIs) were present in SA11-infected cells. The smallest intermediate (precore RI, 45 nm, 220 S) contained the structural proteins VP1, VP3, and VP9 and the nonstructural proteins NS53, NS35, and NS34. A second intermediate (core RI, 60 nm, 310 S) contained the core proteins VP1, VP2, and VP3 and the proteins VP9, NS35 and NS34. The largest RI (single-shelled RI, 75 nm, 420 S) contained the inner shell proteins VP1, VP2, VP3, and VP6 and the proteins VP9, NS35 and NS34. Analysis of the formation and turnover of RIs in infected cells pulse-labeled with 35S-amino acids supports a hypothesis that rotavirus single-shelled particles are assembled in vivo by the sequential addition of VP2 and VP6 to precore RIs consisting of VP1, VP3, VP9, NS35, and NS34. PMID- 2552664 TI - Mutations in 3'-untranslated region of avian sarcoma virus mutant LA46 genome confer the cis-acting temperature-sensitive replication defect. AB - Avian sarcoma virus (ASV) PRCII mutant LA46 carried a unique cis-acting temperature sensitive (ts) defect affecting virus replication. The 3' untranslated region of the LA46 genome contained multiple base substitutions in the direct repeat region and a long insertion. Although these sequences were not present in PRCII, most of these changes were found in various other ASVs and the long insertion shared extensive homology with the so called E region in certain ASV strains. By constructing chimeric retroviruses, the ts defect was mapped to the altered 3'-noncoding region. However, site-specific mutagenization targeted towards a four-nucleotide substitution unique to LA46 failed to implicate these mutations in the ts phenotype. PMID- 2552665 TI - Identification of an enhancer-type sequence that is responsive to Z and R trans activators of Epstein-Barr virus. AB - We studied the regulatory region of the open reading frame BHRF1 of Epstein-Barr virus (EBV) DNA that was responsive to two trans-activators (Z and R) encoded by the BZLF1 and BRLF1, respectively. The 200-bp sequence, nucleotide numbers 53,617 to 53,817 on the EBV map, was sufficient for conferring responsiveness to Z and R. This 200-bp sequence also enhanced expression of the chloramphenicol acetyltransferase (CAT) gene from the simian virus 40 promoter in response to both Z and R, even when inserted downstream of the cat gene. The results indicate that the Z and R response sequence upstream of the BHRF1 has the properties of an enhancer. PMID- 2552666 TI - A physiological correlate of the Pulfrich effect in cortical neurons of the cat. AB - When a swinging pendulum is viewed with a light-attenuating filter before one eye, the pendulum bob is perceived to move in an elliptical path in depth. It is believed that the filter causes this illusion, the Pulfrich effect, by delaying processing of the image in the filtered eye relative to that of the unfiltered eye. We sought a physiological correlate of this effect by studying binocular integration in cortical neurons of cats while they viewed moving stimuli. Special attention was focused on single unit disparity tuning because it is widely believed that depth perception is related to the responses of disparity selective neurons in visual cortex. We found that placing a filter before one of the cat's eyes produced a temporal delay in the cortical response. The temporal delay was always associated with a shift in the neuron's spatial disparity tuning. The observed temporal delays and disparity shifts are comparable with the magnitude of the Pulfrich effect in humans. PMID- 2552667 TI - [Fiber in the diabetic diet]. AB - An increased dietary fibre intake can improve in diabetic patients the compensation of diabetes. The effect of fibre depends on its composition, mode of administration, daily intake and the carbohydrate content of the diet. The reduction of the blood sugar level depends also on the patient's body weight and the degree of compensation of diabetes. PMID- 2552668 TI - [Hematomas of peripheral nerves during traction trauma]. PMID- 2552669 TI - [The artificial pancreas in the diagnosis and intraoperative localization of insulinoma]. PMID- 2552670 TI - [Functional changes in the pituitary-adrenal system during the action of electroacupuncture on patients with spinal osteochondrosis]. AB - A session of low-frequency electroacupuncture (EAP) of the nerve points of the cervicobrachial or lumbosacral regions was performed in 61 patients with vertebral osteochondrosis. An EAP therapeutic effect, analgetic one in particular, was found to depend on cortisol and aldosterone blood levels: reduced levels of the hormones related to EAP procedure appeared associated with a positive therapeutic response. PMID- 2552671 TI - [Mixed metaplastic cancer of the breast]. AB - Basic criteria for making morphologic diagnosis were identified as a result of a study of 4 cases of mixed metaplastic cancer of the breast: segments of the chondroid and osteoid tissue intermittent with structures of infiltrative ductal cancer. Tumor presented as acini-like structures and areas consisting of polymorphous cells with necrosis and calcification. Osteogenesis occurred in some areas. Distant metastases were observed in two cases at months 160 and 127 after treatment. These patients died at months 169 and 134 after treatment as a result of tumor progression. It is recommended that similar treatment be given for mixed metaplastic cancer and infiltrative ductal cancer of the breast. PMID- 2552672 TI - [Diagnosis and follow-up of treatment in early stomach cancer]. AB - Early stomach cancer was identified in 38 patients by application of different diagnostic programs. Follow-up of patients with precancerous gastric pathology proved the most effective in early detection of cancer. The five-year survival rate in radically operated patients with early gastric cancer was 86.5%. PMID- 2552673 TI - [Contents of ACTH, free and bound forms of corticosteroids in blood of patients with stomach cancer]. AB - Levels of ACTH and those of free, protein-binding and total 11 hydroxycorticosteroids were measured in blood plasma in 19 cases with stomach cancer aged 42-68 years. Patients with unaltered pituitary-adrenal cortex links developed non-adrenal hypercorticoidism resulting from decrease in hormone binding to blood proteins. PMID- 2552674 TI - [Ultrasonic examination in tumors of the abdominal cavity and retroperitoneal space in children]. AB - Thirty-two 4 month--5 year old children suffering liver tumors, renal cysts, Wilms' tumor and dermoid cysts of the ovary underwent ultrasonic examination. The data obtained were compared to those of histologic examination of autopsy and resected material. PMID- 2552675 TI - [Pancreatic necrosis after surgical interventions in lung cancer]. PMID- 2552676 TI - [Anti-atherosclerotic properties of eicosapentaenoic acid: clinico-experimental study]. AB - Administration of eicosapentaenic acid (EPA) preparations to patients with type IIb hyperlipoproteinemia brought about a 22-36-percent decrease in blood serum of total cholesterol, triglycerides, beta-lipoproteins, and total lipids. At the same time the administration of EPA preparations was followed by the acquisition of the antiatherosclerotic properties by the patients and healthy donors serum. This could be judged from the reduction of intracellular cholesterol accumulation and intensity of the proliferation of the cells of human aorta intima, inoculated into the culture of the study serum. The data obtained suggest that the antiatherosclerotic properties of the serum are determined to a certain degree by the correlation in it between EPA and arachidonic acid. PMID- 2552677 TI - [Effect of linoleic acid in the diet on arterial pressure, ATPase activity and cyclic nucleotide level in the heart of rats with spontaneous hypertension]. AB - A study was made of the effect of exogenous linoleic acid introduced with feed in the amount of 0.1 cal. % (group I), 9 cal. % (group II) and 16 cal. % (group III) on arterial pressure, Na, K- and Ca-ATPase activity, and on the content of cyclic nucleotides in normotensive Wistar Kyoto rats (NR) and in spontaneously hypertensive Okamoto Aoki rats (SHR). It is established that both deficiency (group I) and excess (group III) amount of linoleic acid in the diet brought about different material changes in the activity of ATPases. Nevertheless, arterial pressure rose in these animals' groups as compared to control (group II). At the same time the degree of arterial pressure rise in NR was higher than in SHR (in percent with respect to the control rats). The content of cAMP and cGMP was appreciably reduced in the animals of the 3 groups (in NR and SHR). PMID- 2552679 TI - [A new type of bread enriches with dietary fibers]. PMID- 2552678 TI - [Anabolic effects of sodium selenite, vitamin E and retabolil in experimental hypotrophy induced by a pesticide]. AB - Forty Chinchilla rabbits of both sexes were examined for changes in some parameters of protein, lipid and trace elements metabolism (total protein, protein fractions, urea, residual nitrogen in blood serum, lipids, total cholesterol, free cholesterol, diglycerides, phospholipids, triglycerides, free fatty acids and the trace elements selenium, iron, zinc and so forth in the liver) during the animals' poisoning with the defoliant magnesium chlorate. A study was made of the effect on these changes of the administration for 3 weeks of sodium selenite (15 micrograms/kg), vitamin E (25 mg/kg) and retabolil (2 mg/kg once a week). It has been established that the combined administration of these agents removes and prevents the changes in protein, lipid and trace elements (hypoproteinemia, dysproteinemia and impairment of the nitrous balance) and lipid metabolism because of the animals' poisoning with magnesium chlorate. PMID- 2552680 TI - [Vitamin D3 in the combined treatment of patient with hypoparathyroidism]. AB - Forty-four patients suffering of hypoparathyroidism were receiving in the complex treatment vitamin D3. Blood calcium and phosphorus, parathyroid hormone, urinary excretion of calcium were determined. The primary clinical effect was manifest on the 3-7 day of treatment; the effect achieved its maximum in most patients by the 2-3 week and in some cases by the 3-6 months of treatment. PMID- 2552681 TI - [Modern achievements in the study of hepatitis A virus and in the development of a vaccine against this infection]. PMID- 2552682 TI - [The role of papilloma viruses in the induction of primary human tumors]. PMID- 2552683 TI - [Epizootic hepatitis A among African green monkeys kept in a vivarium]. AB - The results of observations on the pattern of spread of hepatitis A virus and immune response to it in African green monkeys (Cercopithecus aethiops) kept in the animal house are presented. The infection in the monkeys was found to be characterized by all virological, serological, and biochemical parameters inherent in hepatitis A virus. The results indicated that hepatitis A in monkeys may run both asymptomatic and clinically manifest course, and the spread of infection in the animal house sequentially involves most seronegative animals into the epidemic process. PMID- 2552684 TI - [Antiviral activity of derivatives of aldehydophenols, aldehydo- naphthols and sugar derivatives]. AB - The antiviral activity of aldehydrophenol, aldehydonaphthol derivatives and sugar derivatives was studied. The compounds in doses of 150-500 micrograms/ml were shown to reduce the viral infectious activity by 1.5-3.3 lg TCD50/ml. The degree of virus activity inhibition by the synthesized compounds indicates the relationship of the chemical structure with the antiviral effect of these compounds. PMID- 2552685 TI - [Subunit components of transmissive gastroenteritis virus of swine: preparing purified substances and assessing antigenic properties]. AB - Two different methods were used to prepare subunit components of transmissible gastroenteritis (TGE) virus. The ultrastructure of the peplomer glycoprotein E2 of TGE virus was studied. The presence of detergent NP-40 was shown to prevent the formation of this glycoprotein aggregates. In dialysed preparations, the peplomers are aggregated to form multimer structures, the so-called "rosettes". The orientation of the peplomers in the "rosettes" and in the whole virion was identical. The antigenic activity of viral subunits was studied in guinea pigs. Preparations of isolated peplomers were shown to induce enhanced production of virus-neutralizing antibodies whose mean titre exceeded that in the group of animals immunized with whole virion-based preparations. PMID- 2552687 TI - [Change in the structure of a lipid bilayer during inclusion of influenza virus hemagglutinin in it]. PMID- 2552686 TI - [Macrophage disappearance reaction in mice during an infection caused by Coxsackie B3 virus]. AB - Intraperitoneal inoculation of 2 1/2-3-month-old male BALB/c mice with Coxsackie B3 virus in a dose of 10(7.2) TCD50 was demonstrated to be accompanied by inhibition of the starch-induced exudation of leukocytes into the peritoneal cavity. A decrease in the number of macrophages was observed in 3-day exudates induced at 4 and 21 days but not at 0, 7, and 14 days of infection. The macrophage disappearance reaction showed that the intraperitoneal inoculation of the challenge virus dose (10(7.2) TCD50) was not accompanied at 4-5 hours by changes in the absolute number of macrophages in the pre-existing exudates, but an increase in the absolute number of lymphocytes was observed in the exudates induced at 0, 4, 7, and 14 days of infection, and of polymorphonuclear leukocytes in exudates induced at all the intervals studied. It is assumed that these changes, depending on the period of infection, may be determined by immunological and non-immunological mechanisms. PMID- 2552688 TI - [An outbreak of hepatitis A in imported rhesus monkeys]. PMID- 2552689 TI - [The significance of Ixodes ticks in the southern Far East in the circulation of Powassan virus]. PMID- 2552690 TI - [The effect of 3',5'-cyclic adenylic acid (cAMP) on the growth of bacteria]. AB - The effect of intracellular cAMP level on the growth of bacteria was studied with E. coli AS 1.797, Corynebacterium pekinense AS 1.299 and Bacillus megaterium AS 1.217. The experimental results show that the growth of E. coli AS 1.797 was decreased with increasing cAMP level in cell. With glucose as the sole carbon source, intracellular cAMP is low, and growth of E. coli AS 1.797 was inhibited by exogenous cAMP, but the analog metabolite of cAMP-5'-AMP had no effect on growth. When E. coli AS 1.797 was grown in media containing lactose, maltose and glycerin respectively, intracellular cAMP level is high, and exogenous cAMP did not inhibited the cell growth. When Corynebacterium pekinense AS 1.299 took glucose as carbon source the growth of this bacterium was also inhibited by exogenous cAMP, but the inhibition of cAMP was not specific, its role can be exchanged by 5'-AMP. B. megaterium AS 1.217 cultivated in different carbon source (include glucose) do not contain cAMP in cell, and its growth was not inhibited by exogenous cAMP. 5'-AMP had also no effect on the growth of this bacterium. Thus, the conclusion seems that the cAMP is not essential for the growth of bacteria, and it is a regulator that act a negative regulation in cell growth, but this regulator is of no effect for the growth of B. megaterium AS 1.217. PMID- 2552691 TI - [A 9-week-old infant with congenital cytomegalic inclusion disease and therapy with ganciclovir]. PMID- 2552692 TI - [Short scientific report. Vitamin C lessens superoxide anion(O2)-induced bronchial constriction]. AB - An in vivo model of acute lung damage provoked by free oxygen radicals in guinea pigs was introduced. Bronchial and pulmonary response was induced by tracheal instillation of Xanthine-Oxidase (XO)-Xanthine solution. The response was measured as a decrease of tidal volume during unchanged artificially ventilation. It was found that ascorbic acid pretreatment was effective to diminish the XO induced reaction. PMID- 2552693 TI - [Adequate measurement of artificial mineral fibers and evaluation of the biological effect]. AB - Epidemiological and clinical medical examinations of exposed workers to man-made mineral fibers (MMF) to fix threshold limit values are presupposed by adequate measurements of MMF-dust. As yet are used technics which are unsatisfying. In consideration of the biological conditions in the breathing system a draft to measure MMF dust is introduced and proved. This draft based on the use of two step-gravimetrical measurements together with membranous filter devices and evaluation by an electronic microscope. PMID- 2552694 TI - [The polymerase chain reaction as a sensitive genetic technics method of HIV and HPV diagnosis]. PMID- 2552695 TI - [Infections with human pathogenetic papillomaviruses]. AB - So far, more than 57 types of HPV have been characterized. In contrast, we only know a small number of clinically definable types of warts. Since the individual forms of HPV each have their own oncogenic potential, we should like to get more information on the specific virus-type at least in epidermodysplasia verruciformis and genital lesions. In epidermodysplasia verruciformis, malignant transformation is usually linked to HPV 5, 8, or 14, while HPV 16 and 18 are found in about 70% of pre-cancerous and cancerous genital tumors. PMID- 2552696 TI - [Hemodynamic and medicamentous modification of stimulated atrial natriuretic factor secretion]. AB - The secretion of atrial natriuretic factor (ANF) and its adaptation to pharmacologic and hemodynamic interventions were investigated in 36 patients with sinus rhythm. To provoke standardized secretion of ANF all patients underwent two periods of rapid right ventricular pacing for 4 min with a 15 min interval. Immediately after the first pacing eight patients received 5 mg verapamil, 10 patients 5 mg atenolol and 10 patients 4 mg molsidomine intravenously. Eight patients remained untreated and served as controls. The amounts of atrial pressure increments due to pacing were identical (70% over basal pressure) in all patients. After molsidomine, but not after the other drugs, basal right atrial pressure was lowered. In controls the secretion of ANF due to the second stimulation was significantly (2.5-fold) larger than the secretion induced by the first stimulation. In patients receiving verapamil the secretion response after the second pacing was blunted. Atenolol did not affect the release of ANF. After molsidomine the upward regulation of the ANF secretion rate - seen in controls - was abolished. Thus, the myoendocrine cells are capable for a fast upward regulation of their ANF secretion rate after repeated stimuli. Verapamil directly blocks stimulated ANF secretion, whereas beta-blockade shows no effect. Molsidomine seems to impair enhanced ANF release by lowering basal atrial pressure. PMID- 2552697 TI - [The mechanism of the reinforcing effect of opiates]. PMID- 2552698 TI - [Effect of the local administration of 5,7-DHT and 6-OHDA into the neocortex on the learning and exploratory behavior of rats in an open field]. AB - On Wistar rats characteristics were studied of investigating behaviour in the open field, of learning of conditioned food-reinforced reaction and also of BA and their metabolites content in various brain structures under local intracerebral injections of specific neurotoxins; 6-hydroxydopamine (6-OHDA) and 5,7-dihydroxytryptamine (5,7-DHT), abolishing correspondingly catecholaminergic and serotoninergic terminals. Bilateral injection of 6-OHDA in the neocortex led to a weakening of rats investigating activity in the open field and to an increase of the time of fulfillment of the forming of conditioned food-reinforced reaction. Administration of 5,7-DHT was accompanied by an increase of the investigating behaviour in the open field and a reduction of the duration of the forming of conditioned reaction. Administration of 6-OHDA to the neocortex caused a lowering of catecholamines level in the frontal area of the neocortex and the hippocampus. Analogous administration of 5,7-DHT elicited simultaneously with a deep level lowering of 5-HT and its metabolite in these structures, a change of catecholamines content which testifies to a lesser specificity of the neurotoxin 5,7-DHT in comparison with 6-OHDA. Structures lesion of serotoninergic and catecholaminergic systems of the frontal cortex and the hippocampus brought about by a local administration of 6-OHDA and 5,7-DHT in the neocortex was accompanied by differently directed changes in animals behaviour. PMID- 2552699 TI - Cystosarcoma phylloides in an adolescent female. AB - Cystosarcoma phylloides is an exceedingly rare breast lesion in adolescent females. The tumour contains both stromal and epithelial elements. Benign, malignant and intermediate forms have been described. Complete simple excision is adequate for therapy in all variants. An additional case treated by wide local excision is presented and treatment is discussed. PMID- 2552701 TI - [Rare pancreatic tumors from the viewpoint of our own patient sample]. AB - Four less common tumours of the pancreas are being presented with histological findings (malignant haemangiopericytoma, small cell type anaplastic carcinoma, microcystic cystadenoma, dysontogenetic cyst) from our own patient data. The discussion includes general considerations about preoperative staging and grading as well as the differential diagnosis of the kind of tumours. PMID- 2552700 TI - [Tetrahydrocannabinols in hair of hashish smokers]. AB - The study investigates the presence of tetrahydrocannabinols in the head hair and the pubic and axillary hair. The hair samples were obtained from hashish smokers. The concentrations were determined by radioimmunoassay and reflect total tetrahydrocannabinols and metabolites. The values found ranged from 0.4 ng/mg hair up to 3.8 ng/mg hair. The presence of the drug in the hair samples was also demonstrated by GC/MS. PMID- 2552702 TI - [Does colposcopic and/or cytologic detection of HPV infection without signs of dysplasia require treatment, for example by cryotherapy, laser or trichloroacetic acid (Masviya et al.)?]. PMID- 2552703 TI - The effect of dietary crude fibre content on the frequency of post partum agalactia in the sow. AB - Agalactia (MMA) occurs more or less in all sow herds. In an experiment comprising 14 pairs of full sibs, a low energy sow diet (L = 9.9 MJ ME/kg) was compared with a normal diet (N = 11.5 MJ ME/kg). The incidence of agalactia in the sows was lower in group L than in group N. The weight loss during lactation was higher for L sows, mainly due to feed refusals. The content of dry matter and fat in the milk was somewhat lower in group L. Piglet performance was not affected by the crude fibre content of the sow diet. PMID- 2552704 TI - Reptilian viruses: isolation of parvovirus-like particles from corn snake Elapha guttata (Colubridae). AB - Parvovirus-like particles of about 30 nm in diameter were isolated from the corn snake (Elapha guttata) showing pneumonia. The DNA containing virus replicated in IgH2 cells at 30 degrees C inducing nuclear inclusion bodies. PMID- 2552705 TI - A comparison of three different regimens of infectious bursal disease vaccination in chickens. AB - Five days old progeny chicks from breeders which have received primary and booster doses of live infectious bursal disease vaccine, demonstrated precipitating antibodies unlike progenies from single dose vaccinated breeders. All chicks from the two different groups of breeders were however seronegative at 22 days of age, despite vaccination at 7 or 14 days of age. Post vaccination seroconversion was first noticeable at 32 days in the group of chicks vaccinated at 7 days. Post challenge mortalities was significantly lower (P less than 0.05) and organ lesions very significantly minimized (P less than 0.01) in 7 day old vaccinated group than in 14 or 28 days old vaccinated chicks. These results showed that early (7 days) IBD Vaccination was superior to vaccination at 14 or 28 days, in terms of antibody response and protectivity against mortalities and bursal lesions. PMID- 2552706 TI - Lymphosarcoma development in sheep experimentally infected with bovine leukaemia virus. AB - Twelve sheep were experimentally infected with a phytohemagglutinin (PHA) treated short term culture of lymphocytes from a cow naturally infected with BLV at the PL stage. Five of 12 (42%) BLV infected sheep had histologically confirmed lymphosarcoma 10-16 months after infection. The PBL's were increased to leukemic levels 3-21 weeks before death due to lymphoblastic leukemia. Lymphocyte proliferation and appearance of immature lymphocytes and lymphoblastic cells in the blood were a characteristic feature of tumour development following inoculation with an Australian strain of BLV. In contrast to a number of previous studies the peripheral lymph nodes of all infected sheep were clinically normal throughout the experimental period but at death gross tumours were evident in the mesentric lymph nodes and the heart in all cases. All the other lymph nodes, liver, spleen, kidney and lung were histologically infiltrated with lymphoid tumour cells. Gross tumours were present in the abomasum (1 out of 5) in the urinary tract (2 out of 5) and in the uterus (1 out of 2). The majority of the tumour cells isolated from the various tissues were centroblastic demonstrating that the malignant leukemia in experimentally infected sheep was of a multicentric centroblastic type. The central nervous system was not involved in any case. PMID- 2552707 TI - Crane hepatitis herpesviruses. AB - Comparative studies were performed on crane herpesviruses obtained from two different enzootics in Austria and France. The examined viruses appear to be identical in their physico-chemical properties, morphology and biological reactions in ovo. The crane viruses are tentatively classified as beta herpesviruses. Crane herpesvirus antisera produced in rabbits reacted in cross neutralization tests with each other and with a herpesvirus obtained from a bobwhite quail. No reactivity was observed with other avian herpesviruses and with human herpes simplex virus type 1 and 2. PMID- 2552708 TI - Polyomavirus infection in budgerigars (Melopsittacus undulatus): clinical and aetiological studies. AB - In order to get insight into the aetiology of French Moult (FM) and Budgerigar Fledgling Disease (BFD), and to determine relationships between the two diseases, 298 budgerigars from 49 different breeding flocks were examined. From all birds with BFD and from several birds with FM, viruses were isolated which produced characteristic cytopathic changes in chicken embryo fibroblasts. They were insensitive to chloroform treatment, and their replication was inhibited in the presence of 5-iododeoxyuridine. One of these isolates, from a bird exhibiting clinical signs of BFD, was determined by biochemical and biophysical methods to be a polyomavirus (BFDV). Nestling budgerigars 3 to 10 days of age, were inoculated with this BFDV isolate. They developed an acute systemic disease with high mortality rates, similar to naturally occurring infections. In this regard, BFDV differs markedly from all the other polyomaviruses known until now which only cause clinically asymptomatic, persistent infections in their natural hosts. Most of the birds examined with clinical signs of BFD or FM exhibited low titers of antibodies neutralizing the BFDV isolate, whereas in clinically healthy birds from flocks that never had any problems with FM or BFD, no antibodies against BFDV could be detected. On account of the results of our experiments described here we suspect that FM might be a milder, more protracted form of a BFDV infection. PMID- 2552709 TI - DNA restriction analysis of equine adenovirus serotype I. AB - The three equine adenovirus strains isolated in different locations showed a similar cleavage pattern with HindIII and the DNA homology among the strains was confirmed by Southern blot hybridization. The three strains revealed differences in cleavage patterns with BamHI, EcoRI and PstI, suggesting the presence of DNA polymorphisms among equine adenoviruses. PMID- 2552710 TI - Isolation of a poxvirus from a sparrow (Passer domesticus). AB - From liver, lung, and kidney of a dead sparrow with signs of conjunctivitis a cytopathic agent was isolated in chicken embryo cell culture which was identified as a poxvirus by electron microscopy. From its growth characteristics in avian and mammalian cell culture it was concluded to belong to the genus avipox virus. PMID- 2552711 TI - [Isolation of hemolytic cultures of Bordetella pertussis intended for the production of acellular pertussis preparations and the characterization of their biological properties]. AB - The heterogeneity of the population of B. pertussis laboratory strains with respect to the capacity of individual clones to lyse erythrocytes has been established. The complete or partial reduction of the antigens under study in nonhemolytic bacteria has been shown. The use of hemolytic cultures for the preparation of acellular pertussis vaccine makes it possible to increase the content of B. pertussis toxin in these vaccines. PMID- 2552712 TI - [Tenth anniversary of smallpox eradication: Results of epidemiological and virological surveillance and studies in the post-eradication period]. AB - In accordance with recommendations of the Global Commission on the certification of smallpox eradication for the 10-year period after the eradication of this infection, all suspected cases of smallpox have been thoroughly checked up, and in none of them the diagnosis of smallpox has been confirmed. The study of monkeypox in humans has revealed that this zoonosis is spread over a wider area than supposed earlier and covers 7 countries of Equatorial Africa, occurring most frequently in Zaire. In about 70% of cases of monkeypox in human the disease is contracted from animals serving as natural virus carriers and in about one-third of such cases, from humans having monkeypox. The infectivity of humans with monkeypox for persons having close contacts with them is somewhat lower (12.3%) than in smallpox when this characteristic varies from 37% to 88%. Monkeypox in humans may take an asymptomatic course. Some species of tropical squirrels serve as natural virus carriers. These investigations have also resulted in essential corrections being made in understanding the ecology of cowpox virus, another orthopoxvirus pathogenic for man. At least 5 species of rodents have proved to be of interest as natural carriers of cowpox virus. PMID- 2552714 TI - [Herpetic encephalitis: the clinico-virological aspect of its diagnosis]. AB - Eight patients with herpetic encephalitis (HE) and one patient with herpetic meningitis were investigated. The clinical and laboratory investigations (CR, EEG, and others) are described. In all the cases the diagnosis was corroborated with serological and virological tests unraveling the herpes simplex virus. A high-sensitive diagnose the atypical HE cases with subacute and chronic course that is important for correct treatment. PMID- 2552713 TI - [Nosospecific features of the dynamics of immunological parameters in young children with acute respiratory virus infections of varying etiology]. AB - The study of acute viral respiratory infections has shown that in the course of these infections fluctuations in the content of antigen-nonspecific immunocytes and immunoglobulins occur, depending on the etiology of the disease. This dependence should be taken into consideration in the evaluation of the immune status of young children with acute viral respiratory infections. PMID- 2552716 TI - [Development of endoplasmic reticulum and its enzymic marker in cultured mouse spinal neurons]. AB - In an attempt to elucidate the relationship between synapse formation and cell development, the morphology and cytochemistry of the endoplasmic reticulum and its enzymic marker, glucose-6-phosphatase (G-6-Pase), in cultured mouse spinal neurons were investigated ultrastructurally. It was found that in the early period of the development, neurons were characterized by scarceness of organelles; only a few of granular or agranular endoplasmic reticulum and mitochondria were seen. The endoplasmic reticulum and nuclear envelope were packed specifically with G-6-Pase resection product but the product was weak. After a period of culture, most of the neurons had well-developed endoplasmic reticulum, Golgi apparatus, mitochondria and microtubules, etc. The Golgi apparatus was relatively large, having some cisternae associated with vesicles. Either concave of convex face of the saccules was labeled by thiamine pyrophosphatase (TPPase) specifically. GERL, labeled by cytidine monophosphatase (CMPase), was also seen close to the inner or outer face of some Golgi apparatus. The endoplasmic reticulum at this stage was distributed throughout the cytoplasm, including that in dendrites; its enzyme marker (G-6-Pase) localized consistently within the lumen of all endoplasmic reticulum, nuclear space and subsurface cisternae, and frequently in the concave saccules of the Golgi apparatus. After a long-term culture, some neurons became "aged". The endoplasmic reticulum cisternae enlarged and G-6-Pase reaction reduced. Along with the neuronal development, especially maturation of the endoplasmic reticulum and its enzymic marker, synapse formation was begun at the neuropile area. The axo-dendritic synapses always occurred between the axonal terminals and dendrites where the endoplasmic reticulum had showed positive G-6-Pase reactions. Considering the fact, it suggests that the appearance and change of these specific enzymes may be related to the maturation of the neurons in vitro, and also related to the synapse formation between neurons. PMID- 2552715 TI - [The biological effect of sodium butyrate (NaBT) on SGC-7901 cells]. AB - Changes of (Na+-K+)-ATPase activity, cAMP and fibronectin (FN) content and cell surface microvilli were studied cytochemically, immunocytochemically and scanning electron microscopically on human stomach Glandular carcinoma (SGC-7901) cells treated with NaBT(2.5 mM). It was found that NaBT not only inhibited cell growth but also remarkably decreased the activity of cell surface (Na+-K+)-ATPase of SGC 7901 cells. Note worthy was that, in comparison with the untreated tumor cells, the increase of the intensity of intracellular cAMP and FN immunofluorescence in NaBT-treated tumor cells was striking. Moreover, in contrast to untreated tumor cells, the cell surface of NaBT-treated tumor cells showed more smooth and fewer microvilli under SEM. That NaBT may induce differentiation of SGC-7901 cells through inhibition of (Na+-K+)-ATPase activity and modulation of cellular cAMP and FN content was discussed. PMID- 2552717 TI - Converting enzyme inhibitors in the treatment of elderly hypertensives. AB - Angiotensin-converting enzyme inhibitors are gaining acceptance as safe and effective agents for treatment of hypertension. Addressed in this review of the available literature are the questions whether they are effective in lowering blood pressure in the elderly, whether their effects are age-related, and what effects, if any, do they have on morbidity and mortality in geriatric hypertension. PMID- 2552718 TI - The effects of enalapril on hypertension and quality of life. A large multicenter study in Belgium. AB - Belgian general practitioners studied the effect of enalapril over a period of 6 months on hypertension and quality of life in 4988 patients, whose hypertensive condition remained uncontrolled by previous treatments and/or in whom this previous drug regimen was not tolerated. Blood pressure was reduced from an average of 170/101 mm Hg at the onset of the trial to 145/85.5 mm Hg after 24 weeks. The treatment with enalapril was well tolerated and tolerance improved remarkably in the patients, whose previous treatment had caused intolerance. Quality of life, determined by means of the Nottingham Health Profile questionnaire, was favourably influenced in the patients participating in this study. PMID- 2552719 TI - Non small cell lung carcinoma (NSCLC) at stage 3. A fourteen years retrospective study of 449 patients. Indications of surgery in a multidisciplinary management. AB - Through a fourteen years retrospective study of all the patients treated at the State University of Liege, we have focused on the survival of 449 patients with, at least, a clinical stage 3 NSCLC disease (NORMS UICC). The whole study was divided in two periods (1972-1978, period 1, and 1979-1985, period 2) because, everything remaining nearly equal as far as the clinical material was concerned, period 2 was characterized by a different therapeutic attitude. Since 1979, the NSCLC stage 3's surgical indications have been widely extended. We have performed surgical resections in patients with NSCLC N2 disease because of positive homolateral mediastinal lymph nodes or with T3 disease. This led the NSCLC stage 3 resection rate to move from 26% in period 1 to 47% in period 2. We report here the effects of such a management on the short and long term survival and the stage 3 NSCLC global prognosis. We noted a modest significant increase in the NSCLC stage 3 global prognosis with a five year survival of 4% and 6% (period 1 and 2 respectively; P = 0.03). The 172 resected patients' outcome did not change (five year survival 11% (P1) and 12% (P2]. Although patients were not randomized, our results suggest that adjuvant therapy should not be added whenever the patient underwent a complete resection. On the other hand, it seems very useful indeed whenever the resection was partial. NSCLC patient at stage 3 who underwent a complete resection achieved a 37% five years survival. PMID- 2552720 TI - Complications of surgery in the treatment of lung cancer: their relationship with the extent of resection and preoperative respiratory function tests. AB - We reviewed our experience with the surgical treatment of lung cancer, in order to investigate if the extent of the pulmonary resection and preoperative respiratory function tests correlate with the frequency of postoperative complications. The records of 292 patients who underwent lobectomy and of 64 who had pneumonectomy were analyzed. Postoperative complications developed in 148 patients. The incidence of bronchopleural fistula and cardiac rhythm disturbances was significantly higher after pneumonectomy than after lobectomy. A significant relationship was found between the preoperative ventilatory function tests and the postoperative frequency of intrapleural air spaces or atelectasis. The 30 day operative mortality was significantly related to the extent of the resection; postoperative deaths due to cardiorespiratory insufficiency were also associated to the severity of the obstructive pulmonary disease. PMID- 2552721 TI - The effect of eicosapentaenoic acid in the treatment of atopic dermatitis. A clinical study. PMID- 2552722 TI - Immunobiochemical aspects of atopic dermatitis. PMID- 2552723 TI - Leukotrienes in atopic eczema. AB - Involved skin in atopic eczema contains elevated levels of the 5-lipoxygenase metabolite of arachidonic acid, leukotriene B4. In addition, leukocytes of atopic eczema patients produce increased amounts of eicosanoids upon immunological challenge. These facts and the biological effects of eicosanoids suggest their involvement in the pathogenesis of cutaneous inflammation in atopic eczema, and may provide a new target for pharmacological treatment of this disease. PMID- 2552724 TI - Effects of protein kinase C activation on cyclic AMP and testosterone production of rat Leydig cells in vitro. AB - The role of protein kinase C in modulation of the endocrine function of rat Leydig cells was studied. Percoll-purified rat Leydig cells were stimulated with hCG, forskolin, cholera toxin, pertussis toxin and 8-bromo-cAMP in the presence and absence of two activators of protein kinase C, 12-0-tetradecanoylphorbol 13 acetate (TPA) or 1-oleoyl-2-acetyl-sn-glycerol (OAG). The two activators had no effect on basal cAMP, but decreased hCG-stimulated, and increased cholera toxin- and forskolin-stimulated cAMP production. Cells pre-incubated with pertussis toxin showed enhanced rate of cAMP production in response to forskolin, but were no more responsive to TPA and OAG stimulation. These findings suggest that protein kinase C activation may on one hand inhibit the LH-receptor and Gs protein coupling and on the other hand inhibit the Gi-protein mediated suppression of adenylyl cyclase activity. TPA and OAG effects on testosterone production were measured in the absence and presence of 8-bromo-cAMP stimulation. TPA enhanced basal testosterone production, but this effect was shifted to inhibition when steroidogenesis was stimulated by 8-bromo-cAMP. The OAG effect on testosterone production was inhibitory throughout the dose-response curve of 8 bromo-cAMP. The basal stimulation of testosterone production by TPA was probably due to a marginal increase of cAMP caused by inhibition of the Gi-protein, since a similar effect was observed by pertussis toxin, and thereafter TPA was without effect on testosterone. The inhibition of stimulated testosterone production by TPA and OAG indicates that protein kinase C activity also affects steroidogenesis at a step(s) beyound cAMP formation. PMID- 2552725 TI - Study on clinical and endocrine characteristics of dexamethasone-suppressible hyperaldosteronism compared with those in primary aldosteronism owing to aldosterone-producing adenoma. AB - The clinical and endocrine characteristics of 12 Japanese patients with dexamethasone-suppressible hyperaldosteronism were compared with those in 49 Japanese patients with primary aldosteronism due to aldosteronoma. The results were as follows: 1. Most of the laboratory data in the two groups were almost the same. 2. The grade of vascular damage in both uncontrolled (3) and well controlled (9) patients with dexamethasone-suppressible hyperaldosteronism did not correlate with blood pressure response. 3. The responsiveness of plasma aldosterone to exogenous ACTH in 6 patients with dexamethasone-suppressible hyperaldosteronism was not different from that in 9 patients with aldosteronoma. Even in 3 well-controlled patients in the former group, the plasma aldosterone response was as low as in all the 3 patients with small aldosteronomas. 4. In 4 patients with small aldosteronomas, plasma aldosterone was continuously suppressed with daily dexamethasone to the same degree as in dexamethasone suppressible hyperaldosteronism. 5. The blood pressure, however, did not improve even in the patients with small aldosteronomas. The possible indistinguishable mechanism in dexamethasone-suppressible hyperaldosteronism and primary aldosteronism with small adenomas and the role of unknown hypertensinogenic steroid(s) other than aldosterone in inducing hypertension in dexamethasone suppressible hyperaldosteronism are discussed. PMID- 2552726 TI - Studies on the subcellular mechanisms mediating the negative estradiol effect on GnRH-induced LH-release by rat pituitary cells in culture. AB - Cultured pituitary cells from adult female rats were treated for 4 or 24 h in the absence or presence of E2 (10(-9) mol/l) with increasing concentrations of the transcription inhibitor actinomycin-D or the translation inhibitor puromycin. During the last 4 h of incubation, LH release was stimulated with 5 X 10(-10) mol/l GnRH. The positive E2 effect observed after 24 h treatment with the steroid was clearly abolished by actinomycin-D at concentrations greater than or equal to 10(-10) mol/l and by puromycin at concentrations greater than or equal to 10(-5) mol/l. These findings indicate that the positive E2 effect on GnRH-induced LH release is fully dependent on intact mRNA and protein synthesis. The negative E2 effect observed after 4 h treatment with the steroid was not affected by actinomycin-D and abolished by puromycin only at concentrations greater than or equal to 10(-4) mol/l. Similar results were obtained, when cells had been treated for 4 h with actinomycin-D or puromycin before the 4 h E2 treatment started. Thus, the negative E2 effect seems to be independent of mRNA synthesis and dependent on protein synthesis to a lesser extent than the positive E2 effect. In an attempt to identify positively the subcellular mechanism via which E2 exerts its negative effect, several steps in the GnRH stimulus secretion coupling mechanism were checked whether or not they are modulated by E2. The negative effects of E2 (10(-9) mol/l) on LH release induced by GnRH (10(-10), 10(-9) mol/l) and by the activators of voltage dependent Ca2+ channels K+ (64 mmol/l) or veratridine (3.3 X 10(-5) mol/l) were comparable to those of the calcium antagonist verapamil (10(-6) mol/l). These findings supported the speculation that E2 might act on the Ca2+ channels. The LH release induced by the Ca2+ ionophores A 23 187 (10(-4) mol/l) or ionomycin (6.6 X 10(-5) mol/l), however, was also significantly reduced by 10(-9) mol/l E2, indicating that the steroid modulated a mechanism secondary to the increase of intracellular Ca2+. Also GnRH (10(-9), 10(-8) mol/l) induced accumulation of [3H]inositol phosphates was not influenced by E2 (10(-9) mol/l) treatment, though the steroid exerted a significant negative effect on the LH release by these cells, indicating that phosphatidylinositol-4,5-biphosphate breakdown is not the point of attack for the estrogen.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2552727 TI - Maturational changes in steroidogenic enzyme activities metabolizing testosterone and dihydrotestosterone in two populations of testicular interstitial cells. AB - The present study examined changes in steroidogenic enzyme activities which metabolize testosterone or dihydrotestosterone between days 21-73 of maturation in Band 2 and Band 3 cells isolated by centrifugation of rat testicular interstitial cells on metrizamide density gradients. 5 alpha-reductase and 17 beta-hydroxysteroid dehydrogenase activities increased progressively in Band 2 and Band 3 cells between days 21-35 of maturation, then both enzyme activities declined to reach low levels in adult Band 2 and Band 3 cells. The significantly higher activities of both enzymes in Band 3, which contains a greater concentration of Leydig cells at each age, suggest their localization in Leydig cells. 5 alpha-androstane-3 alpha- and 3 beta-hydroxysteroid dehydrogenase activities increased in both Band 2 and Band 3 cells between days 21-50 of maturation and remained elevated; however, dihydrotestosterone was metabolized primarily to 5 alpha-androstane-3 beta,17 beta-diol in Band 2 cells, while 5 alpha-androstane-3 beta,17 beta-diol was the major metabolite of dihydrotestosterone in Band 3 cells. These studies suggest that testosterone accumulation during sexual maturation can be influenced by changing patterns of 5 alpha-reductase and 17 beta-hydroxysteroid dehydrogenase activities which metabolize testosterone, and of 5 alpha-androstane-3 alpha- and 3 beta hydroxysteroid dehydrogenase activities which metabolize dihydrotestosterone in both Band 2 and Band 3 cells. PMID- 2552728 TI - Impaired ACTH and beta-endorphin response to sauna-induced hyperthermia in heroin addicts. AB - To evaluate the responses of circulating beta-endorphin, met-enkephalin and ACTH to sauna-induced hyperthermia, 8 male heroin addicts recently admitted to a therapeutic community and 8 age-matched normal subjects were examined. Compared with control subjects, heroin addicts showed 1. A decrease of the basal levels of beta-endorphin; 2. Absence of the normal increase of beta-endorphin and ACTH after sauna; 3. A lower increase in systolic blood pressure. It is concluded that an impairment of the adaptive response to stress may be present in heroin addicts, even after a relatively short drug-free period (14 days). PMID- 2552729 TI - Thalassemic patients are at high risk for transfusion-transmitted cytomegalovirus infections. AB - The prevalence of total as well as IgM antibodies against cytomegalovirus (CMV) was determined by enzyme immunoassay in a group of 287 multitransfused thalassemia major patients aged 5-39 years and in another group of 1,220 healthy controls. A significantly higher prevalence of CMV antibodies was observed in thalassemic patients of all age-groups compared with controls. The prevalence among splenectomized thalassemia patients was higher than among nonsplenectomized thalassemics. It is concluded concluded that patients with thalassemia, especially if splenectomized, are at high risk for transfusion-transmitted CMV infections. The high prevalence of CMV infections might be responsible, at least in part, for the immunological disturbances and the susceptibility to other infections observed in thalassemic patients. On the basis of these results, it is suggested that safe blood should be provided for anti-CMV-negative thalassemics, with priority to anti-HIV-positives and those who are to receive bone marrow transplantation. PMID- 2552731 TI - Personality traits and neurohumoral stress response in healthy young sportsmen. PMID- 2552730 TI - Pyrimidine 5'-nucleotidase acquired deficiency in beta-thalassemia: involvement of enzyme-SH groups in the inactivation process. AB - Pyrimidine 5'-nucleotidase (P5'N) partial deficiency has been described in several hematological disorders and also in the beta-thalassemic trait. To check if the P5'N deficiency in thalassemia was acquired we used thalassemic red cells (from either homo- or heterozygous subjects), whose P5'N activity was lower than in control cells. After separation on a density gradient, activity in lighter cells was similar to controls and less than 50% in denser cells. The most probable mechanism for this faster inactivation involves enzyme -SH groups modification by oxidation and reaction with monofunctional aldehydes produced by membrane lipid peroxidation. In vitro challenge of thiol enzymes as pyruvate kinase (PK), adenylate kinase (AK) and P5'N with increasing concentrations of GSSG, hexanal (HEX) and 4-hydroxynonenal (HNE), showed that HNE is the most powerful among the enzyme inhibitors tested and that P5'N activity is a more sensitive index of -SH groups damage, when compared to PK and AK. PMID- 2552732 TI - Effects of opioid receptor antagonists on morphine-induced stupor in rabbits. PMID- 2552733 TI - Endocrine patterns before and after examination stress in males and females. AB - Plasma levels of cortisol, adrenocorticotropin (ACTH), and growth hormone (GH), were determined by radioimmunoassay in medical students just before (Exp.1) plasma values of cortisol were higher than their control values in both sexes, statistically significantly, however, only in males. The difference of cortisol behavior between the sexes was not significant. The males' ACTH values were lower than their control levels, but there was no such difference in the females. Plasma levels and differences of growth hormone varied greatly in both sexes. After the examination (Exp.2) plasma values of cortisol were unaffected compared with their control values in both sexes. The ACTH values were lower than the control values in both sexes, significantly though only in the males. The GH values were higher than control values in the males but not in the females. The results are in line with previous observations suggesting that psychological stress is associated with slightly different hormonal effects in males and females. PMID- 2552734 TI - High cardiovascular reactivity to psychological challenge associated alternatively with high density of beta-adrenergic receptors or with high resting level of catecholamines. PMID- 2552735 TI - Adverse effects on the brain in connection with isoflurane-induced hypotensive anaesthesia. AB - As a marker of brain cell injury, adenylate kinase (AK) was measured in cerebrospinal fluid (CSF) in 10 patients given anaesthesia with isoflurane induced hypotension for corrective surgery of dentofacial deformities. Nine out of 10 patients displayed a marked increase in CSF-AK postoperatively compared with preoperative values. The postoperative mean value displayed a 400% increase compared to the corresponding preoperative value. This difference was statistically significant (P = 0.001). The rise in CSF-AK was most probably the result of an enhanced efflux of AK into CSF subsequent to a presumed hypoxic injury to brain cells. PMID- 2552736 TI - Host-supplied connective tissue as a guide for proliferating tumor cells in human tumor xenografts. AB - The morphologic alterations of 7 human tumors of different origin and various histologic types, heterotransplanted to athymic nude mice, were investigated in the present study. Constant patterns of histologic and ultrastructural changes were observed in all 7 tumors. Following the initial dying of most inoculated tumor cells, host-supplied inflammatory cells invaded the xenografts and phagocytosed necrotic tumor cells. Fibroblasts which vivaciously produced collagenous material invaded the xenografts and built up solid strands of connective tissue which tightly contacted surviving tumor cells. These stands were used as guide-rails for ingrowing blood vessels. Immediately after their immigration, the first mitoses of tumor cells occurred in close proximity to capillary-conducting strands of connective tissue resulting in a revival of tumor cell proliferation near to the fibrous cords and a spreading of newly formed tumor cells along the strands of connective tissue. These results point to the host-supplied connective tissue as playing an important role for tumor proliferation and local tumor expansion. PMID- 2552737 TI - Endocytosis in the rat retinal pigment epithelium. AB - Endocytosis in the retinal pigment epithelium (RPE) of rats was studied using horseradish peroxidase, microperoxidase and ferritin tracers. Tracer uptake was mediated by coated pits and coated vesicles. Coated pits formed at two discrete regions at the RPE plasma membrane: that portion of basal membrane directly opposing Bruch's membrane, and at the bases of the apical lamellae and villi. Two populations of coated vesicles were identified and distinguished by size, location and function. Large coated vesicles (91.8 +/- 14.7 nm in diameter) were located near the cell surface and incorporated tracer. Small coated vesicles (64.5 +/- 15.7 nm diameter) located more deeply within the cell were not tracer labeled, and were often fused with the endoplasmic reticulum or the Golgi apparatus. Observations of the endocytic pathway in rat RPE cells are presented. Tracer was also found in organelles of the lysosomal system, e.g. the multivesicular body, but was not identified in the smooth endoplasmic reticulum or Golgi apparatus. PMID- 2552738 TI - Dense-cored vesicles and unusual lamellar bodies in type III gustatory cells in taste buds of rabbit foliate papillae. AB - Some type III cells in taste buds of rabbit foliate papillae have greatly increased numbers of dense-cored vesicles. Such cells also contain unusual lamellar bodies that resemble those in alveolar type II cells; they consist of alternating dense and light layers with a periodicity of about 4.4 nm. The precise relationship between the vesicles and the lamellar bodies is unknown. PMID- 2552739 TI - Increased incidence of thymoma in Chinese myasthenia gravis: possible relationship with Epstein-Barr virus. AB - Thymectomy was carried out for treatment of myasthenia gravis in 27 unselected Chinese patients and thymoma was found in 13 of them. This 48% incidence of thymomas is two to three times greater than in Japanese and European patients, respectively. The reason for the higher incidence of thymomas observed in Chinese patients may be related to the presence of the Epstein-Barr virus genome in thymoma. Furthermore, all of the thymomas in our patients were lymphoepithelial and histologically resemble nasopharyngeal carcinoma and undifferentiated carcinoma of the salivary gland. Both these tumours are closely linked to the Epstein-Barr virus and in Hong Kong, nasopharyngeal carcinoma is the third commonest cause of death from malignancy. We recommend early thymectomy for patients with myasthenia gravis particularly in geographical areas where there is a high incidence of nasopharyngeal carcinoma and undifferentiated carcinoma of the salivary gland. PMID- 2552740 TI - Prenatal Rh-immune prophylaxis with 300 micrograms immune globulin anti-D in the 28th week of pregnancy. AB - As immune globulin anti-D given in the immediate post partum period fails to prevent the development of anti-D antibodies in about 1.5-2% of women at risk, probably as a result of feto-maternal bleeding during pregnancy, 300 micrograms of immune globulin anti-D was administered to 609 Rh-negative women in the 28th gestational week. Three hundred and forty-six had Rh-positive babies, and were given additionally 200 micrograms anti-D post partum. Of these, 291 had an antibody screen test done 10 months after delivery. No anti-D antibodies were found. The test results of the study group were compared with those of a control group of 354 women who did not receive prenatal immune globulin, but otherwise the same examinations and treatment. In this group, 322 had a screen test performed 10 months post partum or in their next pregnancy, when 1.8% had anti-D antibodies. The difference in immunization incidence between the groups was significant (p less than 0.05). There was no difference between the groups regarding the number of women with fetal erythrocytes in serum after delivery, but the number of fetal erythrocytes found was significantly lower in the group receiving prenatal prophylaxis (p less than 0.001). No adverse effects were found in the infants exposed to anti-D prenatally. PMID- 2552741 TI - Twenty years' experience of treating gestational trophoblastic neoplasia in Sweden. AB - Experience gathered during the last 20 years in the treatment of gestational trophoblastic neoplasia (GTN) is presented. Fifty-eight cases were treated at the Karolinska Hospital during the period of 1966-86. This number accounts for approximately 50% of the cases in Sweden during this period of time. Thirty-four patients developed their malignancy after a molar pregnancy. Thirty-three patients were nonmetastatic and of the 25 metastatic cases, 6 patients were staged to a high-risk group. Chemotherapy was given to all patients except one who had a primary choriocarcinoma in an ectopic pregnancy. Surgery was performed in elective cases or when chemotherapy failed. Surgical procedures were also performed in a few emergency situations. Since 1978 all patients were started on methotrexate--folinic acid ad modum Goldstein. When this medication failed, either the CHAMOMA (vincristine, methotrexate--folinic acid, actinomycin D, melphalan, adriamycin) or later the EMA/CA (etoposide, methotrexate--folinic acid, actinomycin D, cyclophosphamide, adriamycin) regimen ad modum Bagshawe was given. An overall remission rate of 96.5% was achieved. The minimum period of follow-up was 2 years. The 2 patients who died (in 1966 and 1982) were referred to our department in a late stage. Five patients had a late recurrence and all but one were successfully treated. PMID- 2552742 TI - Corneal topography and elasticity in normal and keratoconic eyes. A methodological study concerning the pathogenesis of keratoconus. AB - One eye of 27 patients with keratoconus and 37 normals were examined by photokeratoscopy and topographic pachometry to obtain informations about the corneal shape and thickness profile. The progressive alteration of corneal topography in keratoconus may be based on elastic deformation. Through a distention, predominantly of the central area, increasing large differences between central and peripheral thickness appear. Simultaneously the area of the corneal surface increases into a more elliptical corneal shape. In keratoconus relative to normals, the study demonstrates, an alteration of, as well as a correlation between, the corneal shape and thickness profile in accordance with the elastic hypothesis. Based on corneal topography determination, intraocular pressure measurements and a derived elastic theory the corneal tissue mass (M), tissue strength (Young's Modulus in steady state Y(s) , and the ocular rigidity (E) were determined. Y(s) and E was found to be significantly decreased, whereas M seems to be equal in keratoconus compared with normals. Histologically cornea consists of an amorphous matrix in which the collagen fibers are embedded. E seems to reflect the immediate elastic response of the collagen fibers (Y(i], which, however, may be relaxed in steady state. Y(s) seems to reflect the elastic response of the matrix, which consists of matrix compression and sliding of collagen fibers. Thus the biomechanical alteration of cornea in keratoconus may be introduced by increased sliding fo collagen fibers due to reduced attachment to Bowman's layer and altered synthesis of the matrix substance. PMID- 2552743 TI - Improvement of modern treatment and outcome in childhood epilepsy in Asia. AB - Epilepsy and epileptic syndromes are one of the major pediatric neurological diseases in Taiwan, R.O.C. In 1984 we investigated 38 elementary schools in Taichung city, in the middle-west part of Taiwan. Among 57,944 school-aged children, 388 had suffered from at least two episodes of afebrile seizures occurring separately over a two-week period. The period prevalence rate was 0.67%. In our clinic, once seizures are diagnosed, the seizure types are then classified as well as possible, in order to develop a rational approach to treatment. We commonly use carbamazepine for partial seizures and some generalized seizures, low dose ACTH (10 I.U.) for infantile spasms, valproic acid for absence seizures and myoclonic seizures, clonazepam for atonic-akinetic and myoclonic seizures, and phenobarbital for young children with generalized seizures. In the last three years, we have used a ketogenic diet for akinetic atonic seizures and Lennox-Gastaut syndrome; we have also used temporal lobectomy for complex partial seizures since 1981. Owing to the marked increase in the medical understanding of epileptic seizures and syndromes, and models of treatment, there is no question that children with seizure disorders are better off today than they were ten years ago. PMID- 2552744 TI - Increased in vitro leukotriene B4 production by stimulated polymorphonuclear cells in Kawasaki disease. PMID- 2552745 TI - [Effect of clonidine on the schedule-controlled performance action of two benzodiazepine receptor agonists]. AB - Effect of clonidine, a presynaptic adrenoceptor agonist, on the schedule controlled performance action of two benzodiazepine receptor agonists was assessed in rats in a fixed-ratio (FR6) procedure. Diazepam (0.03 approximately 1.0 mg/kg), an agonist of both type I and II benzodiazepine receptors, decreased the FR performance in a dose-dependent manner. Clonidine (10 micrograms/kg) significantly potentiated the action of diazepam when the dose of diazepam was increased to 0.1 mg/kg. However, clonidine did not show any potentiating effect on the mild depressive action of CL 218,872, a special agonist of type I benzodiazepine receptor until its dose was increased to 10 mg/kg. The Potentiating effect of clonidine on the depressive action of diazepam was significantly antagonized by yohimbine. Bicuculline (3.0 mg/kg) significantly antagonized the depressive action of diazepam (1.0 mg/kg). Our results suggest that the depressive action of benzodiazepines on the function of central noradrenaline system may be one of the benzodiazepine sedative mechanisms, which may be related to the type II benzodiazepine receptors. PMID- 2552746 TI - In situ effects of histaminergic agents on adrenergic neurotransmission in rat mesenteric microcirculation. AB - Experiments were performed to study in situ the effects of histamine and some H1- or H2-agonists and antagonists on the contractions of the rat mesenteric methaarterioles (RMMA) induced by 0.6 nM noradrenaline (NA) applied locally or by electrical stimulation (ES). All drugs tested (cimetidine--300 nM, the others - 30 nM) were also applied locally 30 s before the contractile stimulus. NA and ES strongly contracted the blood vessels. Of all drugs studied only diphenhydramine (DPH) evoked a slight contraction of RMMA. Histamine markedly inhibited the contractile responses to NA and ES. Similar effects were exerted by 2-(2 aminoethyl)thiazole (2-AET) its inhibitory action on the ES-induced contractions being weaker than that of histamine. Dimaprit (DMP) only tended to inhibit the RMMA contractions. DPH abolished the NA-induced contractions while cimetidine (CIM) did not change the effects of NA and ES. The results suggest that H1 receptors are mainly involved in the modifying action of histaminergic drugs on the adrenergic neurotransmission in RMMA. PMID- 2552747 TI - Spontaneous EEG paroxysmal activity, behavioural and endocrine changes in male rats subjected to different regimens of locomotion. AB - The changes in the excitability of the GNS of male rats were investigated following the influence of long-term (6 months) regimens of locomotion: running exercise and relative immobilization. The excitability was evaluated by the changes in EEG paroxysmal activity and in some behavioural states (alertness, exploration and quiet wakefulness). The quantitative alterations of the features mentioned were correlated with the serum levels of several hormones (LH, testosterone, ACTH, non-specific glucocorticoids) in terms to provide a complete assessment of the functional state of the CNS. The inference was drawn, that long term exercise elicits adaptive changes resulting in an increase of the activation threshold of the CNS. Long-term restriction of the locomotion leads to an increase of the sensitivity of the nervous system, i.e. the activation threshold of the CNS was found to be lowered. PMID- 2552749 TI - Lactate transport in isolated mouse muscles studied with a tracer technique- kinetics, stereospecificity, pH dependency and maximal capacity. AB - Lactate transport across the sarcolemma of isolated mouse muscles was studied with a 14C tracer technique. The cellular tracer uptake could be inhibited by unlabelled L-lactate (and pyruvate) and to a lesser extent by D-lactase. The stereospecific fraction had a Km of 3.5 mM, and made up 50% of the total transport. The tracer uptake was unaffected by 0.05 mM DIDS and 0.2 mM amiloride, but was inhibited by cinnamate (Ki = 8 mM) and PCMBS (Ki = 0.8 mM). With high concentrations of the latter inhibitor compounds or with high concentrations of unlabelled L-lactate, the tracer uptake was inhibited 80%, which indicates that the main part of the transport involves facilitated diffusion. The remaining fraction (20%) was non-saturable, reduced at high pH, and could not be inhibited; it is probably mediated by diffusion of undissociated lactic acid. Lactate transport was pH-dependent, which is consistent with a lactate-H+ symport. The maximal transport capacity, as calculated from the pH changes measured with pH sensitive micro-electrodes while the lactate gradient was 30 mM, was 11.8 mmol kg 1 min-1 (pH 6.2). PMID- 2552750 TI - A model for the fast 4-aminopyridine effects on amphibian myelinated nerve fibres. A study based on voltage-clamp experiments. AB - The effect of 4-aminopyridine (4-AP) on the potassium currents in the node of Ranvier in myelinated nerve fibres was investigated with the voltage-clamp technique. The potential and time dependence of the currents in solutions with a high potassium concentration (114.5 mM) were studied. The block of the tail current was found to be less than that of the corresponding current during a potential step. Also, the time-course of the tail current was modified. In order to explain these findings, kinetic models of the 4-AP action were constructed and analysed numerically. A simple four-state model, in which 4-AP interacts only with open channels and where the binding is diphasically potential dependent, was found to account for the effects. PMID- 2552748 TI - Vasoactive intestinal peptide (VIP) may reduce the removal rate of cytosolic Ca2+ after transient elevations in clonal rat lactotrophs. AB - The prolactin-producing rat anterior pituitary GH4C1 cells possess Ca2+-activated K channels which are activated by physiological elevations of the cytosolic Ca2+ concentration even at membrane potentials more negative than the normal level of about -50 mV. Whole-cell current recordings showed a marked outward tail current following depolarizing voltage steps to 0 mV from a holding potential close to the normal membrane potential. The half-time of this tail current was about 1.3 s after a 4-s depolarization step. The GH4C1 cells also possess voltage-activated Ca channels, and we conclude that this tail current is a Ca2+-activated K+ current for the following reasons: (1) The reversal potential for the tail current was close to the K+ equilibrium potential for a range of transmembrane K+ gradients. (2) The tail current was blocked by a Ca2+ antagonist, and the voltage dependence of this current closely mirrored the voltage dependence of the isolated Ca2+ current. The time-course of the decline of the tail current thus reflects the removal rate of the Ca2+ entering the cytosol through voltage dependent Ca channels during the depolarizing voltage step. VIP stimulates prolactin secretion from GH4C1 cells, and this peptide prolonged the half-time of the tail current by about 47% in 63% of the cells. This indicates that VIP may prolong the transient cytosolic Ca2+ elevations following the action potentials in these cells. Such a mechanism might be an important factor for the control of the cytosolic Ca2+ level, and hence hormone secretion. PMID- 2552751 TI - Sympathetic and noradrenaline effects on C-fibre transmission: single-unit analysis. AB - Single afferent unmyelinated fibres were dissected from the otherwise intact sural nerve in anesthetized rabbits. The sympathetic trunk could be stimulated via electrodes implanted through the abdomen. The response in single C fibres was elicited by electrical stimulation in the cutaneous innervation area of the fibre. Sympathetic stimulation (8 Hz, 1 ms pulses, 5 mA for 60 s) increased the latency in all tested C fibres (2.0% +/- 0.8%, mean +/- SD, n = 17). In 48% of the units the amplitude of the action potential decreased (26.4% +/- 12.3%) during sympathetic stimulation. Infusion of noradrenaline (5 micrograms min-1) increased (7.7% +/- 4.1%) the latency in all units and increased (36.9% +/- 29.8%) the amplitude of 25% of the units. The effects of sympathetic stimulation and noradrenaline infusion were blocked by pre-treatment with phentolamine (3 mg kg-1 i.v.). The results suggest that catecholamines change the membrane properties of unmyelinated fibres. PMID- 2552753 TI - Examination of HeLa cell contamination of human cell lines derived from primary hepatomas using glucose-6-phosphate dehydrogenase and lactate dehydrogenase isozymes. AB - Isozyme patterns of glucose-6-phosphate dehydrogenase (G6PD) and lactate dehydrogenase (LDH) in human cell lines derived from primary hepatomas were compared with those in HeLa cells. Some cell lines derived from primary hepatomas having type B G6PD showed one or two isozymes of LDH. On the other hand, HeLa cells having type A G6PD showed four LDH isozymes. These findings suggest that not only G6PD, but also LDH may be useful for the detection of HeLa cell contamination of a culture in some cases. PMID- 2552752 TI - Hepatitis B virus markers in patients with schistosomiasis, liver cirrhosis and hepatocellular carcinoma in Khartoum, Sudan. AB - Markers of hepatitis A and B virus were tested in 88 adult Sudanese subjects in Khartoum, Sudan. The subjects consisted of 25 control hospitalized patients, 21 volunteer blood donors, 23 patients with hepatosplenic schistosomiasis, 13 patients with liver cirrhosis and 6 patients with hepatocellular carcinoma (HCC). Antibody to hepatitis A virus was detected in 96% of the total. Hepatitis B surface antigen (HBsAg) was positive in 4, 24, 22, 31, and 67% of the subject groups, respectively. Antibody against hepatitis B core antigen (HBcAb) of undiluted serum was positive in 60, 57, 65, 77 and 83%, and there was no difference in incidence among the groups. It was positive in 200X diluted serum in 4, 24, 17, 23 and 60%. HBsAg and HBcAb (200X) were detected more often in HCC patients than in the control subjects (p less than 0.01). Hepatitis B virus is an important factor in the etiology of HCC in the Sudan. PMID- 2552754 TI - Near infrared emission of singlet oxygen generated in the dark. AB - Singlet oxygen generation is reported from (1) enzymatic reaction and (2) electron transfer reactions of the superoxide anion measured directly with an ultrasensitive near-IR emission spectrophotometer by monitoring the O2(1 delta g) ---O2 (3 sigma g-) transition at 1268 nm. Near-IR emission spectra from the myeloperoxidase and lactoperoxidase enzymatic systems show only emission of singlet oxygen at 1268 nm. The lipoxygenase/Na-linoleate enzymatic reaction exhibits two emissions, 1268 nm and 1288 nm. The latter emission is identified as originating from a peroxy radical. Spectral and kinetic data giving evidence of singlet oxygen generation is obtained from the reaction of potassium superoxide solubilized by 18-crown-6-ether in acetonitrile with a series of organometallic coordination compounds. PMID- 2552755 TI - Localization of the luminol-dependent chemiluminescence reaction in human granulocytes. AB - The granulocyte luminol-dependent chemiluminescence (CL) reaction is linked to the enzyme myeloperoxidase reacting with products of the respiratory burst activation. The results presented in this paper, show that the light generated in granulocytes originate both from intracellular and extracellular reactions; however, depending on the stimulus used the one or the other will dominate the activity measured. Furthermore, lysosomal fusion is proposed to be required for the intracellular CL reaction. PMID- 2552756 TI - Phagocytosis induction of chemiluminescence and chemoattractant increased superoxide anion release from activated human alveolar macrophages in asthma. AB - Human alveolar macrophages (AM) were demonstrated to generate reactive toxic derivatives of oxygen in many pulmonary disorders. These cells are involved in local inflammation which characterizes bronchial asthma. In the present work, we studied the ability of stimulated macrophages from healthy volunteers, and asthmatic patients to generate oxygen species in vitro. AM obtained by bronchoalveolar lavage were purified by adherence. The production of oxygen species was measured by luminol-enhanced chemiluminescence (CL) after challenge with opsonized zymosan. The maximal values were significantly (p less than 0.03 and p less than 0.01) higher in AM from asthmatics than in AM from healthy subjects. A significant correlation (p less than 0.01) was observed between maximal value of CL and the severity of asthma as assessed by the clinical score. But, no difference was observed between AM from asthmatics in a stable state and healthy subjects. On the other hand, assays for superoxide anion generation emphasized the activation state of these macrophages stimulated by formyl peptides. PMID- 2552757 TI - A chemiluminescent method for the measurement of pregnanetriol-3 alpha glucuronide in human diluted urine. AB - Pregnanetriol-3 alpha-glucuronide (PTG) is the majority urinary metabolite of 17 hydroxyprogesterone (17OHP) and it typically increases in the commonest form of congenital adrenal hyperplasia (CAH), due to 21 hydroxylase deficiency. We developed a simple chemiluminescent immunoassay for the direct measurement of PTG in diluted urine in order to avoid the preliminary hydrolysis and extraction steps that are usually employed in gas-liquid chromatographic methods. The immunogenic complex PTG-bovine-serum-albumin was used to induce the formation of specific antibodies in New Zealand rabbits. In addition, PTG was conjugated to aminoethylethylisoluminol and the resulting tracer was characterized by mass spectrometry and used to monitor the immunological reaction. The characteristics of the antibody were determined with regard to specificity and sensitivity. The precision of the assay method was also established. PTG excretion was studied before and after the ACTH stimulation test (1 mg synthetic ACTH i.m.) in 11 normal women and in one subject affected by CAH due to 21-hydroxylase deficiency. PTG levels well correlated with 17OHP plasma concentrations both under basal and stimulated conditions, in normal women as well as in the patient affected by CAH. PMID- 2552758 TI - Malaria, Epstein-Barr virus, and the genesis of lymphomas. PMID- 2552759 TI - Correction of a chest wall deformity utilizing latissimus dorsi with a turnover procedure. AB - Infraclavicular hollowing and the abnormal anterior axillary fold are often seen in patients with Poland's syndrome and related conditions. The operative procedure described in this article corrects these deformities. The procedure uses the latissimus dorsi island flap. The inserted muscle is moved anteriorly and sutured between the deltoid and the biceps. Then the muscle flap is twisted between its origin and where it is inserted. The muscle fibers act like a pivot and the thickness of the flap is increased, which enables us to achieve a round anterior axillary fold. The muscle flap fills the infraclavicular region and softens the deep hollow area. PMID- 2552760 TI - Modulation of IL-1-induced collagenase production in articular chondrocytes by pertussis toxin. AB - The effects of pertussis and cholera toxins on interleukin-1 (IL-1) stimulated collagenase production from rabbit articular chondrocytes were investigated. Cholera toxin (50-1000 ng/ml) had no significant effect on IL-1-stimulated collagenase production. Pertussis toxin gave a 50-100% inhibition of collagenase activity induced by submaximal IL-1 concentrations. However, pertussis toxin had little effect on collagenase activity induced by maximal concentrations of IL-1. Optimal inhibitory effects were observed with 5-10 ng/ml of toxin. PMID- 2552761 TI - Topical anti-inflammatory activity of DuP 654, a 2-substituted 1-naphthol. AB - Recent work suggests that one of the common biochemical characteristics of skin inflammatory diseases such as psoriasis is altered arachidonic acid metabolism with elevated levels of prostaglandins and leukotrienes. DuP 654, a 2-substituted 1-naphthol, is an exceptionally potent inhibitor of 5-lipoxygenase. DuP 654 was tested in various models of skin inflammation and was found to be potent at inhibiting edema induced by the topical application of arachidonic acid, tetradecanoyl phorbol acetate or the calcium ionophore A23187. DuP 654 was also effective in a murine model of contact sensitivity. DuP 654 was effective at reducing the numbers of infiltrating polymorphonuclear leukocytes in AA and TPA induced edema. These data, taken together, suggest that DuP 654 may be effective in treating human skin diseases. PMID- 2552762 TI - Leukotriene (LT) receptor antagonists. Heterocycle-linked tetrazoles and carboxylic acids. LY203647. AB - LY171883, 1-[2-hydroxy-3-propyl-4-[4-(1H-tetrazol-5-yl)butoxy]phenyl] ethanone, is an orally active antagonist of LTD4- and LTE4-induced responses in a variety of test systems. We prepared a new series of LT antagonists based on a proposed model of LY171883 binding to the LTE4 receptor in which the n-propyl and tetrazole moieties of LY171883 occupy those parts of the receptor to which the C1 C5 chain and the cysteinyl carboxyl of LTE4 bind, respectively. The new compounds have an acidic function corresponding to the glycine carboxyl of LTD4 linked through a heterocyclic group which is proposed to bind to the LTD4 receptor where the cysteinyl glycine amide bond of LTD4 binds. LY203647, 1-[2-hydroxy-3-propyl-4 [4-[2-[4-(1H-tetrazol-5-yl)butyl]-2H- tetrazol-5-yl]butoxy]phenyl] ethanone, showed good LTD4 antagonist activity with a suitable pharmacologic and toxicologic profile and has been chosen for clinical evaluation. PMID- 2552763 TI - In vitro inhibition of arachidonic acid metabolism by two novel retinoid analogs. AB - Ro 23-6457, (all-E)-3,7-dimethyl-9-[2-(trifluoromethyl)-6-(nonyloxy)phenyl]-2, 4,6,8- nonatetraenoic acid, and Ro 23-2895, (all-E)-9-[2-(nonyloxy)phenyl]-3,7 dimethyl-2,4,6,8-nonatetraen oic acid, are two novel retinoid analogs which exhibit antiinflammatory activity in both the developing and the established rat adjuvant arthritis models [8]. Here we investigated the effect of these two compounds on the production of arachidonic acid (AA) metabolites in two in vitro test systems [i.e., Ca2+ ionophore A23187 (I)-stimulated resident rat peritoneal macrophages (MO) and cytokine-stimulated human dermal fibroblasts (HDF)]. Both compounds, Ro 23-6457 and Ro 23-2895, significantly inhibited the release of 14C AA metabolites and the production of LTB4, PGE2, and 6-keto-PGF1 alpha in I stimulated MO, at concentrations of 1-33 microM. Both compounds also inhibited the production of PGE2 in HDF stimulated by either rhuIL-1 alpha or huTNF alpha at concentrations of 1 x 10(-5) to 1 x 10(-7) M. Ro 23-2895 was also a potent inhibitor of IL-1-induced collagenase production in rheumatoid synovial cells (IC50 approximately 1 to 2.5 x 10(-8) M). The inhibitory profile of these novel compounds in these cell systems is therefore similar to that of other known antiinflammatory retinoids (e.g., all-trans- and 13-cis-retinoic acid). Inhibitory effects such as those described here might in part contribute to the antiinflammatory activity of these compounds in vivo. PMID- 2552764 TI - The pharmacology of arachidonic acid-induced rat PMN leukocyte infiltration. AB - Arachidonic Acid (AA) injected into a hindpaw of Lewis rats produces high levels of tissue myeloperoxidase (MPO), a biochemical marker for PMN leukocytes. Treatment with a corticosteroid (prednisolone) or dual 5-LO/CO inhibitors of AA metabolism (phenidone, SKF 86002) produced dose-related inhibition of AA-induced elevations in paw tissue MPO levels. In contrast, administration of high pharmacologic doses of selective cyclooxygenase inhibitors (indomethacin, ibuprofen, naproxen), anti-histamine/serotonin agents (cyproheptadine, chlorpheniramine) or an anti-arthritic gold compound (auranofin) produced only slight or moderate effects. Thus, AA-induced hindpaw inflammation is a useful method for determining pharmacologic effects of 5-LO/CO inhibitors on PMN leukocyte infiltration in vivo. PMID- 2552765 TI - Ultrastructural analysis of leukocytes recruited into the guinea pig lung by LTB4 aerosol. AB - Two groups of naive guinea pigs were exposed to different LTB4 aerosol solutions: LTB4/saline and LTB4/BSA. Control animals were exposed to saline alone and saline/BSA. The LTB4/BSA aerosol was the most effective in recruiting leukocytes, with the maximal numbers of cells in the lung tissue at 6 hrs. Light microscopy/TEM revealed marked differences in eosinophil morphology in saline/BSA and LTB4/BSA aerosol treated animals. Occasional eosinophils in the submucosa of controls, had few pseudopodia, few cytoplasmic vesicles, and intact granules. In LTB4 treated animals, there were numerous eosinophils in the bronchial mucosa and submucosa with numerous pseudopodia, numerous cytoplasmic vesicles, and light crystalline cores. PMID- 2552766 TI - Comparison of LTB4- and C5a-stimulated chemotaxis of isolated human neutrophils: difference revealed by cell migration in thick filters using the multiwell cap procedure. AB - Under comparable conditions (90 min incubation in 2% albumin buffer) using 3 micron pore cellulose nitrate filters and the multiwell cap procedure (Evans et al., Bioscience Reports 6:1041, 1986), C5a was more potent than LTB4 as a chemoattractant (EC'50 s = 0.5 and 4 nM) and caused 5 times as many cells to completely traverse the filter. During the 90 min incubation, no cells traversed the filter in the absence of chemoattractant. The cap assay was modified to study migration of cells within the filter. During the 30 min incubation, EC50 values for C5a and LTB4 were comparable (1.2 and 1.6 nM) in causing cells to enter the filters but C5a was superior in causing the cells to move as measured by the mean distance of migration (EC'50 s = 0.25 and 1.8 nM). Our studies support the view that LTB4 acts primarily to cause cell adhesion (and penetration) of the endothelium and that C5a plays a major role in cell migration (McMillan and Foster, Agents and Actions 24: 114, 1988). PMID- 2552767 TI - Granulocyte-macrophage colony-stimulating factor (GM-CSF) and opsonization synergistically enhance leukotriene B4 (LTB4) synthesis induced by phagocytosis in human neutrophils. AB - Normal human blood neutrophils were studied for their capacity to synthesize leukotriene B4 (LTB4) and its omega-oxidized metabolites after phagocytosis of zymosan. Phagocytosis of serum-opsonized particles led to a higher release of LTs than did unopsonized zymosan. The most striking effect of phagocytosis was observed when neutrophils were primed with granulocyte-macrophage colony stimulating factor (GM-CSF): opsonization and GM-CSF synergistically increased LTB4 synthesis by neutrophils. PMID- 2552768 TI - The in vivo antiinflammatory effects of (E)-2,6-bis(1,1-dimethyl-ethyl)-4-[2-(5 methyl-1H-pyrazol-3-yl)ethenyl ] phenol (PD 127443) a novel dual inhibitor of 5 lipoxygenase and cyclooxygenase. PMID- 2552769 TI - The effect of leukotriene-B4 receptor antagonist, SC-41930, on acetic acid induced colonic inflammation. AB - SC-41930, 7-[3-(4-acetyl-3-methoxy-2-propylphenoxy)-propoxy]-3,4-dihydro-8-p ropyl- 2H-1-benzopyran-2-carboxylic acid, is a potent in vitro leukotriene-B4 (LTB4) receptor antagonist. LTB4 levels are elevated in colonic tissue of inflammatory bowel disease (IBD) patients which may account for the high degree of neutrophil (PMN) infiltration. The guinea pig acetic acid-induced colonic inflammation model has characteristics of IBD including PMN infiltration, edema, ulceration and necrosis. The model was used to evaluate the effect of SC-41930. SC-41930 was given orally, 30 min before and after intrarectal administration of 3% acetic acid. The PMN marker enzyme, myeloperoxidase, was measured along with histological evaluation to assess inflammation. Both parameters showed significantly less inflammation in SC-41930 treated animals with an oral ED50 of 20 mg/kg. These study results with an LTB4 receptor antagonist indicate a role for LTB4 in colonic inflammation and that an LTB4 receptor antagonist may be beneficial for treatment of IBD. PMID- 2552770 TI - Rheumatoid arthritis synovial fluid phospholipase A2 activating protein (PLAP) stimulates human neutrophil degranulation and superoxide ion production. AB - Rheumatoid arthritis is characterized by excessive eicosanoid production, and phospholipase enzymes are the rate limiting step in eicosanoid synthesis. We have shown previously that cells from patients with rheumatoid arthritis express enhanced phospholipase A2 enzyme activities. Recently, we have isolated a phospholipase A2 activating protein termed PLAP from rheumatoid synovial fluid. This novel human protein shares biochemical and antigenic similarities with melittin, a bee venom phospholipase activating protein. Because melittin has been shown to induce neutrophil degranulation and superoxide formation, and because exuberent release of lysosomal enzymes and superoxide have been implicated in the pathogenesis of rheumatoid arthritis, we examined the role of PLAP on inducing these neutrophil functions. PMID- 2552771 TI - Chondrocyte activation by interleukin-1: synergism with fibroblast growth factor and phorbol myristate acetate. AB - Exposure to synovial factors or purified interleukin-1 (IL-1) induces the production of prostaglandin E2 (PGE2) and the neutral proteinases (NP) collagenase, gelatinase and stromelysin by lapine articular chondrocytes. Having frequently found our partially purified synovial preparations to elicit this process of chondrocyte activation more strongly than recombinant IL-1, Phadke's report of synergism between IL-1 and fibroblast growth factor (FGF) intrigued us. In our hands, basic FGF (1 ng/ml-1 micrograms/ml) did not activate chondrocytes but, in a dose-dependent manner, enhanced the production of PGE2 and NP by chondrocytes exposed to IL-1 alpha or IL-1 beta (1-10 U/ml). Further examination determined that the basic FGF was a better synergist than acidic FGF. In view of reports that FGF activates protein kinase C, we tested whether phorbol myristate acetate (PMA) could substitute for FGF as a synergist. Not only did it do so, but PMA alone (0.1 ng/ml-100 ng/ml), unlike FGF, provoked the production of PGE2 by chondrocytes. The Ca2+ ionophore A23187 could not substitute for FGF in enhancing induction of the NP. Using a cDNA probe, we confirmed that the synergistic effects of both FGF and PMA upon IL-1 mediated collagenase induction, were associated with an increased abundance of collagenase mRNA. PMID- 2552772 TI - Cycloheximide inhibits the induction of collagenase mRNA in chondrocytes exposed to synovial factors or recombinant interleukin-1. AB - Synoviocytes secrete "chondrocyte activating factors" (CAF) which, like recombinant interleukin-1 (IL-1), induce the synthesis of collagenase by cultures of articular chondrocytes. Enzyme synthesis is preceded by the appearance of collagenase mRNA some 3-5 hours after exposure of the chondrocytes to CAF or IL 1. Cycloheximide (CX) inhibits the appearance of this message in a dose-dependent manner. At a concentration of 5 micrograms/ml inhibition by CX is completely reversible, with superinduction being observed in certain experiments. Identification of the newly synthesised proteins which are required for collagenase mRNA induction would greatly advance our understanding of collagenase gene expression. PMID- 2552773 TI - Effect of tumor necrosis factor on granule release and LTB4 production in adherent human polymorphonuclear leukocytes. AB - Tumor necrosis factor (rTNF) has previously been shown to induce PMN chemotaxis, stimulate PMN adhesion to vascular endothelium and stimulate hydrogen peroxide secretion from PMNs adhered to biological surfaces. We investigated the activity of both rTNF alpha and rTNF beta on adherent and suspension cultures of human PMNs. rTNF alpha selectively stimulated the release of the specific granule in a dose dependent manner. Exocytosis of the specific granule was measured with an enzyme-immunoassay for lactoferrin and a radioassay for vitamin B12-binding protein. Adherent PMNs released up to 60% of the total lactoferrin content of the cells with no increase in myeloperoxidase (MPO) secretion when stimulated with 0.1-10 nM rTNF alpha. The PMNs in suspension cultures also selectively released the specific granule, although total release was reduced suggesting that adherence of PMNs increased their ability to respond to physiological stimuli. When PMNs in suspension cultures or adherent cells were stimulated with rTNF alpha, no LTB4 production was detectable, yet the cells retained the ability to synthesize LTB4 when stimulated with calcium ionophore A23187. Neither rTNF alpha or rTNF beta stimulated the release of the azurophilic granule, measured by the secretion of MPO and neutrophil elastase activity. These results suggest that a function of rTNF alpha and rTNF beta on PMNs is the release of the contents within the specific granule. PMID- 2552774 TI - Enhancement of human neutrophil leukotriene synthesis by human granulocyte macrophage colony-stimulating factor. PMID- 2552775 TI - Development and application of a membrane receptor assay for leukotriene B4. AB - 5(S),12(R)-dihydroxy-6-cis-8,10 trans-14-cis-eicosatetraenoic acid (LTB4) is a potent inflammatory mediator generated by human cells. A receptor assay using membranes from cultured HL-60 cells has been developed to quantitate LTB4 with a range of sensitivity from 10 pg to 5 ng. The initial metabolite of LTB4, 20-OH LTB4, has a cross reactivity of 28% while other lipoxygenase products do not significantly compete. This assay has been used to study ionomyocin-induced formation of LTB4 by human neutrophils. The use of membranes from HL-60 cells for the measurement of LTB4 provides a sensitive and highly selective alternative to radioimmunoassay for the determination of the levels of this important eicosanoid in biological fluids and should be useful in the development of antagonists of the LTB4 receptor. PMID- 2552776 TI - Hypochlorous acid (HOCl) activation of neutrophil collagenase requires cathepsin G. AB - In an effort to understand the mechanism of collagenase activation in inflammation, human peripheral neutrophils were isolated and incubated with the tumor promoter, phorbol myristate acetate (PMA), which induces the neutrophils to degranulate and secrete proteinases. Neutrophil media were then treated with HOCl with or without various proteinase inhibitors then collagenase activity was measured. Added HOCl was able to activate latent collagenase. However, a serine proteinase, cathepsin G, was found to be necessary for collagenase activation to occur by HOCl. The results indicate that cathepsin G is a key mediator in neutrophil collagenase activation and that HOCl under certain conditions leads to the activation of cathepsin G or the stimulation of cathepsin G's ability to activate neutrophil collagenase. PMID- 2552777 TI - [Intraocular fluid antibody quotients following inoculation with two different antigens in experimental herpes simplex virus retinochoroiditis]. AB - Viral retinochoroiditis was induced experimentally by inoculation of herpes simplex virus into the vitreous body in rabbits. The animals were sensitized systemically using 2 pathogens as antigens (herpes simplex virus and toxoplasma gondii) 3 weeks prior to intraocular inoculation. Serum and intraocular fluid were collected 2 weeks after inoculation. The intraocular fluid antibody titers and quotients for these 2 pathogens were then measured to determine the effects of serum antibodies, which are thought to enter the eyes as a result of destruction of the blood-ocular barrier. Experimental criteria for antibody quotients were also determined. Antibody quotients for the etiological virus (herpes simplex virus) ranged from 2 to 20, with an average of 9.7. Those for toxoplasma gondii, the antibody of which is thought to enter the eye from the blood, were all less than 5 with average of 1.9. From these results, it would seem that when the antibody quotient of a pathogen in more than 6 in the intraocular fluid, it is likely to be an etiological organism, while the possibility of infection is very low when the quotient is under 2. More precise studies are required to identify an etiological pathogen when the quotient is 2 6. PMID- 2552778 TI - [Urachal carcinoma producing carcinoembryonic antigen: a case report]. AB - We report a case of urachal carcinoma with elevated serum carcinoembryonic antigen (CEA) level in a 36-year-old man. En bloc resection was performed. The production of CEA was proved histologically. The level of serum CEA returned to the normal value after operation. Seventeen postoperative months the level of serum CEA was noted to be elevated again, and 2 months later symptoms appeared and tumor recurrence was revealed on computer tomographic scan. Radiotherapy and chemotherapy were done without any noticeable response. He died of tumor progression. In our case, serum CEA gave useful information as a tumor marker. It showed specificity for the tumor and helped evaluation of tumor resection as well as detection of tumor recurrence. PMID- 2552779 TI - [Clinical investigation of 7 cases of urachal carcinoma]. AB - Seven cases of urachal carcinoma experience in Yokohama City University in these 10 years are presented herein. The high incidence of local recurrence of this disease and the results of the treatments in the previously reported cases were reviewed. From these experiences, extensive radical surgery including total cystectomy, en bloc removal of entire urachus, pelvic lymphadenectomy, and adjuvant chemotherapy and or radiation are recommended for the treatment of urachal carcinoma. PMID- 2552780 TI - [A case report of adult Wilms' tumor]. AB - A 49-year-old women who presented with right upper abdominal mass and back pain had a nephroblastoma, Wilms' tumor, in the right kidney. After radical nephrectomy the patient was treated with chemotherapy in the form of actinomycin D and vincristine. Despite the intensive chemotherapy, the patient died of respiratory failure by multiple metastasis at chest wall and lung. The pathology and treatment of adult Wilms' tumor were discussed. PMID- 2552781 TI - [A case of signet ring cell carcinoma of the urinary bladder]. AB - A 67-year-old man was admitted for complaint of gross hematuria on April 9, 1986. Cytoscopic examination was revealed broad-base tumor, its size was thumb's head, at the right lateral wall of the bladder. Ultrasonography and computed tomographic scan of the bladder demonstrated no evidence of invasion to adjacent organs. The biopsy showed signet-ring cell carcinoma. Radiography of the digestive organs showed no abnormality. A partial cystectomy was performed. Histologically the tumor was limited up to submucosa, that is pT1b. The localization of carcinoembryonic antigen (CEA) on tissue of our case, using the peroxidase-antiperoxidase (PAP) method, was evidenced. PMID- 2552782 TI - Edward B.D. Neuhauser memorial lecture. Reflections of men and machines from red goggles and spin wobbles. PMID- 2552783 TI - Malignant primary germ cell tumors of the mediastinum: CT features. PMID- 2552784 TI - OPC-88117 suppresses early and delayed afterdepolarizations and arrhythmias induced by cesium, 4-aminopyridine and digitalis in canine Purkinje fibers and in the canine heart in situ. AB - The effects of OPC-88117, a new investigational antiarrhythmic drug, on early and delayed afterdepolarizations (EAD and DAD, respectively) were assessed in vitro in canine Purkinje fibers and in vivo in the canine right ventricle. OPC-88117 had similar electrophysiologic properties to class I antiarrhythmic agents in that it decreased Vmax. OPC-88117 decreased the amplitude and prolonged the coupling interval of DAD induced by acetylstrophanthidin. Likewise, OPC-88117 suppressed EAD induced in vitro by 4-aminopyridine. In vivo, cesium-induced EAD, ventricular arrhythmia, and atrioventricular block were suppressed by OPC-88117. In summary, OPC-88117 suppressed DAD and EAD in vitro and inhibited EAD and triggered activity in the in situ canine heart. PMID- 2552785 TI - Assessment of respiratory exposures during gilsonite mining and milling operations. AB - An industrial hygiene study of the entire United States gilsonite industry was done by the National Institute for Occupational Safety and Health (NIOSH) to evaluate the potential for occupational health problems resulting from exposures to gilsonite and its constituents. Gilsonite is a solidified hydrocarbon substance mined only in northeastern Utah to Colorado. Industrial hygiene samples were collected at four gilsonite mining companies including nine mines and three mills. Gilsonite workers had no measurable exposures to polynuclear aromatic hydrocarbon (PNA) compounds, asbestos fibers, or hydrogen sulfide gas. Several organic gases/vapors and metals were detected in the airborne samples; but, none exceeded the current exposure standards/health criteria of the Mine Safety and Health Administration (MSHA), the American Conference of Governmental Industrial Hygienists (ACGIH), or NIOSH. Gilsonite workers in some job categories were exposed to high levels of dust, exceeding ACGIH nuisance dust recommendations. These dusts, comprised largely of aliphatic hydrocarbons, had a large aerodynamic size distribution with average mass median aerodynamic diameters (MMAD) above 30 microns. PMID- 2552786 TI - Occupational exposure and analysis of microcrystalline cristobalite in mullite operations. AB - Three analytical methods currently used for crystalline free silica determination in occupational exposure samples were evaluated for their applicability to ceramic materials containing synthetic mullite. X-ray powder diffraction is the only method that can be used with sufficient precision and potentially adequate accuracy for occupational exposure monitoring. Personal respirable dust exposure samples were collected in two foundries. The results of exposure evaluations in Plant A showed overexposure to the dust, particularly in the shakeout area. The cristobalite concentration in the respirable dust exceeded that in the original material for this particular area. This may be related to the preheating of molds to more than 1100 degrees C for the pouring of stainless steel castings, causing conversion of some of the colloidal silica binder to cristobalite, and related to high vibration fracture of the material during removal of castings from molds. In Plant B, quartz was sometimes present along with cristobalite in personal samples. Respirable dust levels exceeded the Occupational Safety and Health Administration's (OSHA) permissible exposure limit (PEL) and the National Institute for Occupational Safety and Health's (NIOSH) recommended time-weighted average (TWA) for crystalline free silica, indicating a need for better engineering controls to reduce dust levels. The inadequacy of reference standards currently available for cristobalite analysis in these types of materials is cited. The need for more toxicological research is emphasized. PMID- 2552787 TI - Localization of a "postreceptor" defect in human dilated cardiomyopathy. PMID- 2552788 TI - Nutritional factors in diabetics with and without retinopathy. AB - We used 3-d food-record-keeping techniques to examine nutritional factors in diabetic patients with and without retinopathy. Patients without retinopathy had significantly higher daily intakes of total carbohydrate, water-soluble dietary fibers, insoluble dietary fibers, and glucose than did patients with retinopathy. Also, patients without retinopathy took a significantly lower proportion of their total daily calories as protein. PMID- 2552789 TI - Malignant salivary gland tumors of the paranasal sinuses and nasal cavity. The UCLA experience. AB - Between 1962 and 1985, 35 patients with malignant salivary gland tumors of the paranasal sinuses and nasal cavity were treated with curative intent at UCLA. They were staged according to the American Joint Committee (TNM) classification for squamous cell carcinoma of the paranasal sinuses: 27 of 35 (77%) patients presented with T3-4 disease. Adenoid cystic carcinoma was the diagnosis in 24 (68%) patients, while adenocarcinoma accounted for half of the cases involving the ethmoid sinuses and nasal cavity. Eleven patients were treated with surgery alone, 13 with combined surgery and radiation, and 11 with radiation therapy alone. All patients were followed from 40 to 216 months. Analysis by treatment modality revealed a local control of 18% (2 of 11) in the surgery alone group, 62% (8 of 13) in the combined group, and 9% (1 of 11) in the radiation alone group. The group undergoing combined treatment achieved the highest local control rate despite having a higher proportion of patients with advanced stage and residual disease at the surgical margins. In patients with positive surgical margins, the tumor recurred locally in 4 of 6 (67%) unirradiated patients, compared with 3 of 10 (30%) of those undergoing postoperative irradiation. Patients with adenoid cystic carcinoma had poorer results, with only 25% remaining relapse-free, compared with 45% of patients with adenocarcinoma. The 5 , 10-, 15-, and 20-year actuarial survival for all cases was 73%, 60%, 32%, and 20%, respectively. We conclude that post-operative radiation improves the results of treatment in all patients with malignant salivary gland tumors in this location, compared with the results of surgery alone. PMID- 2552790 TI - Breast carcinoma kinetics. Argyrophilic nucleolar organizer region counts correlate with Ki67 scores. AB - This study assessed the value of argyrophilic nucleolar organizer region (AgNOR) staining as a potential technique for the estimation of cell kinetics in conventional histology sections, in benign and malignant breast lesions. Using a silver staining technique and immunohistochemistry, the authors correlated the numbers of argyrophilic nucleolar organizer regions (AgNORs) and Ki67 scores in 70 breast carcinomas and 27 benign breast lesions. Epithelial cells in fibrocystic disease and fibroadenomas contained a mean of 2.65-6.8 small uniform AgNORs per cell, whereas malignant cells contained 4.6-26.9 frequently highly irregular AgNORs. In benign tissue, Ki67 scores ranged from 0 to 4%; in malignant tumors, Ki67 scores ranged from 3.0 to 98%. The correlation between AgNOR counts and Ki67 scores was highly significant (P less than 0.001). The authors concluded that AgNOR counts performed on routine formalin-fixed paraffin sections furnish significant kinetic information. Furthermore, the difference in AgNOR counts between benign and malignant tumors is such that they may be of diagnostic value. PMID- 2552791 TI - A monoclonal antibody against nerve growth factor receptor. Immunohistochemical analysis of normal and neoplastic human tissue. AB - The expression of human nerve growth factor (NGF) receptor in tumors and normal tissue was investigated with the use of a monoclonal antibody recently developed against that protein. This antibody, NGFR5, reacted strongly with 100% of 25 nerve sheath tumors. Eight of nine pheochromocytomas and three of three paragangliomas also had positive results, but the immunoreactivity was restricted to the sustentacular cell population. Within cells of melanocytic lineage, there was no immunostaining of melanocytes in normal epidermis, whereas 13 of 14 benign nevi had positive results, primarily involving spindled nevocytic structures within the dermis. NGF receptor was scarcely expressed in human melanoma; 9 of 19 melanomas had positive results, but immunoreactivity was generally restricted to rare cells within the larger tumor cell population. Among nonneurogenic mesenchymal tumors, results were generally negative: 0 of 5 chondrosarcomas, 0 of 6 malignant fibrous histiocytomas, 0 of 3 meningiomas, and 1 of 8 leiomyosarcomas were immunoreactive. Carcinomas were variable in immunoreactivity: 12 of 16 squamous cell carcinomas had positive results, whereas adenocarcinomas demonstrated focal, basal epithelial immunoreactivity and neuroendocrine tumors generally had negative results. Among normal tissues, in addition to expected neural immunostaining, NGFR 5 reacted positively with several nonneural cell types, including lymphoidal follicular dendritic cells, myoepithelial cells, vascular adventitia, and basal epithelium of oral mucosa and hair follicles. Antibodies to NGF receptor may play a role in the identification of benign and malignant soft tissue lesions. PMID- 2552792 TI - Lymphokine-activated killer (LAK) cell activity in tumor-infiltrating lymphocytes from non-small cell lung cancer. AB - Tumor-infiltrating lymphocytes (TILs) are often seen in non-small cell lung cancers (NSCCs). Their functional role in the pathogenesis of lung cancer is unknown. The authors studied TILs in 27 patients with NSCC and determined the following: (1) the immunologic phenotype as defined by monoclonal antibodies against various surface markers, (2) activation state as indicated by interleukin 2 (IL-2) receptor expression and the kinetics of proliferation response to IL-2, and (3) the ability to develop lymphokine-activated killer (LAK) type cytotoxicity against both natural killer (NK)-resistant tumor cell targets (M14) and fresh autologous tumor cells. The authors' results show TILs from NSCCs to be a heterogeneous population composed of T-cells, B-cells, monocytes, and NK cells in frequencies similar to those found in peripheral blood lymphocytes (PBLs). TILs demonstrated increased IL-2 receptor expression and a more rapid proliferative response to IL-2 than PBLs, implying activation of TILs by the tumor milieu. Finally, TILs generated cytotoxicity against NK-sensitive (K562) and NK-resistant (M14) cell line targets consistently after in vitro treatment with IL-2 but were less consistent in their ability to lyse fresh autologous tumor cells and less effective than PBL LAK cells in lysing all targets. Comparison with LAK cells generated from normal volunteers suggests that decreased killing of autologous tumor cells only partially results from an inherent resistance to lysis by fresh NSCC targets. It appears, therefore, that tumor cells taken from NSCCs are not readily killed by the immune cells that infiltrate the tumor stroma and that this failure does not result from nonspecific immune deficiency in TILs. PMID- 2552793 TI - Antibodies to an Epstein-Barr virus nuclear antigen synthetic peptide in infectious mononucleosis. Report of two cases. AB - The Epstein-Barr virus nuclear antigen (EBNA-1) contains a region of repeating glycine and alanine amino acids. It has been shown that this region contains a major epitope of EBNA-1. With well-characterized sequential sera from two cases of acute infectious mononucleosis, a specific IgM response was detected to the EBNA-1 synthetic peptide by enzyme-linked immunosorbent assay (ELISA). Conversely, an IgG response was observed in the convalescent phase of the illness with a progressive decline of the IgM antibodies. This response was observed with heterophil-positive and heterophil-negative EBV/IM. The peptide-specific serologic response was confirmed by immunoblotting, the serial serum samples on extracts of EBV transformed B-cells. There was excellent correlation between the antipeptide ELISA and blotting techniques. PMID- 2552794 TI - Breast carcinoma with positive results for melanoma marker (HMB-45). HMB-45 immunoreactivity in normal and neoplastic breast. AB - HMB-45 (melanocytic cell-specific monoclonal antibody) immunoreactivity has been detected by the immunoperoxidase technique in the cytoplasm of 2 of 100 breast carcinomas. In addition, normal breast lobules and ducts were occasionally found to have positive results in 6 of 100 cases. These findings indicate the need to exercise caution in the interpretation of HMB-45 immunoreactivity for immunohistochemical characterization of neoplasms. PMID- 2552795 TI - Signet ring cell sinus histiocytosis. A previously unrecognized histologic condition mimicking metastatic adenocarcinoma in lymph nodes. AB - The axillary lymph nodes in a radical mastectomy specimen from a 70-year-old insulin-dependent diabetic patient contained significantly vacuolated sinus histiocytes. The histologic picture closely resembled metastatic signet ring cell adenocarcinoma. The signet ring histiocytes did not stain with the mucicarmine or periodic acid-schiff stains or any of the immunohistochemical epithelial markers. The differential diagnosis and the possible origin of these vacuolated histiocytes are discussed. PMID- 2552796 TI - Bilateral adrenal hemorrhage as a complication of intravenous ACTH infusion in two patients with inflammatory bowel disease. AB - We report two patients with inflammatory bowel disease in whom intravenous corticotropin therapy was complicated by bilateral adrenal hemorrhage. In one, the initial presentation was acute and unilateral, simulating colonic perforation and/or abscess. The diagnosis was made only at exploratory laparotomy. In the second patient, the presentation was subacute; multiple episodes of adrenal hemorrhage occurred subsequent to the course of corticotropin, and ultimately, hypoadrenalinism developed. Pertinent clinical and computed tomographic findings are reviewed. PMID- 2552797 TI - Erosion of Marlex band and silastic ring into the stomach after gastroplasty: endoscopic recognition and management. AB - We report three patients who developed erosion of a Marlex band or a silastic ring that was used to reinforce the stoma after vertical stapled gastroplasty. The patients were symptomatic and the diagnosis was made by endoscopy. After endoscopic removal of the foreign body, symptoms resolved in two patients. To our knowledge, this is the first report of successful removal, by endoscopy, of an eroding mesh or a ring. PMID- 2552798 TI - Carcinoma of the appendix presenting as biliary colic. PMID- 2552799 TI - Serpin receptor 1: a hepatic receptor that mediates the clearance of antithrombin III-proteinase complexes. AB - Antithrombin III (ATIII) clearance from blood occurs by redistribution into the extravascular compartment and by binding to the endothelial surface. When, however, ATIII reacts with a proteinase such as alpha-thrombin, the complex is rapidly cleared from the circulation (half-life is approximately five minutes) by a receptor present on hepatocytes. This receptor binds a number of other serine proteinase inhibitors that are members of the class designated as the "serpins." ATIII, alpha 1-proteinase inhibitor, heparin cofactor II, and alpha 1 antichymotrypsin proteinase complexes bind to the same hepatic receptor, now designated as serpin receptor 1. Proteinase complexes with alpha 2-antiplasmin, another member of the serpin class, do not bind to serpin receptor 1. Recent studies suggest that the specificity of the receptor for serpins may reside in the so-called D helix (nomenclature based on the structure of alpha 1-proteinase inhibitor). The presence of ATIII on the surface of endothelial cells offers a unique mechanism for regulating proteinases formed during coagulation. Since this ATIII is probably associated with heparin-like substances and exists in a high affinity state, the inhibitor rapidly binds proteinases such as alpha-thrombin. Once the complex forms, its affinity for heparinoids is decreased compared with ATIII, allowing the complex to dissociate from the endothelial surface for rapid clearance by the liver. PMID- 2552800 TI - Normalization of LDL receptor function by lymphocytes of patients with heterozygous familial hypercholesterolemia after treatment with plasma cholesterol lowering agents. AB - Low density lipoprotein (LDL)-dependent growth of mitogen-activated lymphocytes, inhibited in their capacity to synthesize cholesterol endogenously, can be used as an assay of functional receptors for LDL. Using this technique, abnormalities can be detected in circulating lymphocytes obtained from patients with familial hypercholesterolemia (FH). Functional lymphocyte LDL receptor activity was decreased in patients with heterozygous FH. Following treatment with the specific inhibitor of cholesterol synthesis, lovastatin, alone or in combination with a bile acid-binding resin, there was increased expression of functional lymphocyte LDL receptors in five of nine patients. Plasma LDL cholesterol levels decreased in all nine patients. Three other patients who were only studied while receiving therapy also manifested increased expression of functional lymphocyte LDL receptors. The degree of improvement in plasma LDL cholesterol did not predict the effect on lymphocyte LDL receptor function. Longitudinal studies indicated that an increase in functional LDL receptor activity could be observed with 4 weeks of therapy and persisted for at least 18 months on continuous treatment. These results provide direct evidence that therapy with lovastatin and a bile acid-binding resin can lead to increased expression of functional LDL receptors by lymphocytes in the majority (eight of 12) of patients with heterozygous FH. PMID- 2552801 TI - Clinical disorders of renal tubular phosphate transport. PMID- 2552802 TI - Urinastatin (Kunitz-type proteinase inhibitor) reducing cisplatin nephrotoxicity. AB - The authors investigated the reductive effects of a Kunitz-type proteinase inhibitor, urinastatin, on the nephrotoxicity seen in lung cancer patients treated with cisplatin by measuring N-acetyl-beta-D-glucosaminidase (NAG) activity and beta 2-microglobulin (BMG) content in 24 hour urine, creatinine clearance, blood urea nitrogen (BUN), serum creatinine, uric acid, and BMG as factors of nephrotoxicity. In control patients treated with anticancer drugs containing cisplatin but no supplemental urinastatin, the 24 hour urine NAG and BMG levels increased more than three-fold over the pretreatment levels, 3 days after anticancer therapy, respectively. Creatinine clearance significantly decreased and levels of BUN, serum uric acid, and BMG in control patients significantly increased over the corresponding pretreatment levels, 3 days after anticancer therapy. However, supplemental urinastatin reduced abnormalities in levels of all these factors 3 days after therapy. These results suggest that supplemental urinastatin protects from cisplatin-induced nephrotoxicity, especially proximal tubular damage. PMID- 2552803 TI - AIDS peripheral neuropathy. AB - A young male IV drug abuser with multiple AIDS risk factors and positive HIV serology presented with acute onset of painful urinary retention as a result of a parasympathetic mononeuropathy. Because no other explanation could be found for his illness, despite a careful search for the most likely etiologic agents, the authors propose that his symptom complex may have resulted from infection with the AIDS virus. A review of the current literature relevant to the peripheral neuropathy associated with AIDS is presented. PMID- 2552804 TI - Pulmonary beta-adrenergic receptors and response do not mature precociously in growth-retarded rabbit fetuses. AB - Growth-retarded infants have a reduced risk of pulmonary morbidity. We used a naturally occurring model of growth retardation in rabbits to test the hypothesis that reduced risk might be related to precocious maturation of the alveolar beta adrenergic response system in runted neonates. We confirmed that the weights of the fetuses were significantly different, depending on uterine position, as predicted by this model. However, we found no evidence of either increased beta adrenergic receptor concentration or cyclic adenosine monophosphate generation in the smaller fetuses. These results indicate that reduced fetal size in this model of growth retardation does not result in accelerated maturation of alveolar beta adrenergic responses in neonates. PMID- 2552805 TI - The renin-angiotensin system during pregnancy in chronically instrumented, conscious rats. AB - Whether the renin-angiotensin system is activated during rat gestation is controversial. Therefore we serially assessed plasma renin activity in unrestrained, chronically instrumented conscious rats during pregnancy and the postpartum period. Plasma renin activity was 3.26 +/- 0.30, 2.80 +/- 0.31, and 2.70 +/- 0.26 ng.ml-1.hr-1 on gestational days 6, 12, and 20, respectively. When the same rats were studied after delivery, plasma renin activity was 1.87 +/- 0.29, 1.81 +/- 0.09, and 2.31 +/- 0.35 ng.ml-1.hr-1 on postpartum days 3, 6, and 11, respectively. Levels measured during pregnancy were significantly greater than in the postpartum period (p less than 0.05 or less than 0.01). We then evaluated potential functional consequences of the renin-angiotensin system in gravid rats. Near term, renal hemodynamics fall from the peak levels of midgestation; we tested whether angiotensin II mediates this apparent vasoconstriction. Captopril (1.5 mg/kg, 1.5 mg.kg-1.hr-1) was acutely administered to lower circulating angiotensin II. The drug produced an 80% inhibition of angiotensin I pressor response, a tenfold elevation in plasma renin activity, but caused the same degree of mild renal vasodilation in rats whether they were virgin or pregnant. We also tested whether prior occupancy of receptors by endogenous hormone or receptor downregulation mediates the attenuated pressor response to angiotensin II observed during late pregnancy. Acute administration of captopril failed to augment refractory pressor responsiveness. Chronic treatment with enalaprilat (2.0 mg.kg-1.day-1 by osmotic minipump) also did not restore pressor responsiveness. But, in our hands, chronic administration of enalaprilat most likely failed to lower plasma angiotensin II. In summary, we suggest that during rat gestation (1) the renin-angiotensin system is activated, (2) angiotensin II does not mediate the apparent renal vasoconstriction observed near term, (3) prior receptor occupancy by endogenous hormone is not responsible for the attenuated pressor response to angiotensin II, and (4) long-term treatment with enalaprilat can produce hypotension without reducing plasma concentrations of angiotensin II. PMID- 2552806 TI - Gestational trophoblastic disease and use of oral contraceptives. PMID- 2552807 TI - Localization of gonadotropin binding sites in human ovarian neoplasms. AB - The binding of human luteinizing hormone and human follicle-stimulating hormone to ovarian tumor biopsy specimens from 29 patients was analyzed. The binding sites for human luteinizing hormone were demonstrated in one tumor of epithelial origin (mucinous cystadenoma) and in one of sex cord-stromal origin (theca cell tumor). The binding sites for human follicle-stimulating hormone were found in three tumors of epithelial origin (serous cystadenoma and mucinous cystadenoma) and in two of sex cord-stromal origin (theca cell tumor and theca-granulosa cell tumor). The surface-binding autoradiographic study revealed that the binding sites for gonadotropins were localized in the stromal tissue. The results suggest that gonadotropic hormones may play a role in the growth and differentiation of a certain type of human ovarian neoplasms. PMID- 2552808 TI - Differences between white and Chinese populations in human leukocyte antigen sharing and gestational trophoblastic tumors. AB - The prevalence of gestational trophoblastic tumors varies widely among different populations: it is lowest in whites (3 to 6/100,000) and highest in Chinese (68 to 202/100,000). This observation suggests that the origin of the disease is different in the two populations. To test this hypothesis, we examined couples in whom the woman developed a gestational trophoblastic tumor in a white population (Pittsburgh) and a Chinese population (Taiwan) for sharing of human leukocyte A, B, DR, and DQ antigens, which we consider markers for sharing of major histocompatibility complex-linked recessive genes affecting both embryogenesis and carcinogenesis. No human leukocyte antigen sharing occurred between partners in Pittsburgh, but there was significant human leukocyte antigen sharing in Taiwan. The latter couples shared human leukocyte antigen B (p less than 0.04) and human leukocyte antigen DQ (p less than 0.007) and shared three or more human leukocyte A, B, DR, and DQ antigens (p less than 0.02) significantly more frequently than did normal couples. However, there was no increased sharing of any specific human leukocyte antigen allele. These findings support the hypothesis that gestational trophoblastic tumors occur on a sporadic basis in whites and on a genetic basis in Chinese. PMID- 2552809 TI - The "normal" brain. "Abnormal" ubiquitinilated deposits highlight an age-related protein change. AB - Known morphologic changes that characterize "normal" brain senescence are insufficient to explain the widespread, age-related decline of psychomotor functions. We report that the heavily ubiquitinilated deposits can be consistently detected by immunohistochemistry in the normal senescent brain. Immunostaining of hippocampal sections from aged brains with an anti-ubiquitin antibody was unrelated to neurofibrillary degeneration or senile plaque formation. In contrast, ubiquitin deposits were not detectable in brain sections from neurologically and neuropathologically normal young individuals who had died of nonneurological causes. This finding shows an unrecognized protein change in the normal aged brain. PMID- 2552810 TI - Quantitative immunohistologic assessment of lymphocyte populations in the pulmonary inflammatory response to intratracheal silica. AB - Immunogold-silver staining was used to identify T lymphocytes, T lymphocyte subsets, and B lymphocytes in lung tissue from mice injected intratracheally with silica, titanium dioxide, or saline alone. Morphometric quantitation revealed a marked influx of T lymphocytes in the silica-treated animals during the first 3 weeks after injection. The relative numerical density of these cells remained elevated when compared with saline-treated controls throughout the 12 weeks of the experiment. Cells expressing the CD4 and CD8 antigens were both increased in number, with the former accounting for approximately two-thirds of the T lymphocytes. An increased number of B lymphocytes was also apparent from 6 weeks after treatment with silica. The T lymphocyte response preceded the development of significant pulmonary fibrosis by several weeks. No lymphocyte response was observed in the lungs of mice injected with nonfibrogenic titanium dioxide. These observations are consistent with the hypothesis that lymphokines secreted by T lymphocytes play a role in the pathogenesis of silicotic inflammatory lesions and their progression to fibrosis. PMID- 2552811 TI - Ceruloplasmin reduces the adhesion and scavenges superoxide during the interaction of activated polymorphonuclear leukocytes with endothelial cells. AB - The plasma protein, ceruloplasmin, has been implicated as an anti-inflammatory agent, although this property has not been demonstrated unequivocally in vivo. The role of this protein in an in vitro system of cultured endothelial cells and polymorphonuclear leukocytes (PMNs) was investigated. One of the initial steps in an inflammatory response is increased adhesion between PMNs and the endothelial lining of the blood vessels. The results showed that ceruloplasmin interferes with this process and reduces the number of phorbol myristate acetate-activated leukocytes that adhere to endothelium. Preincubation of either the activated PMNs or the endothelium with ceruloplasmin did not produce the same results, suggesting that the continuous presence of ceruloplasmin is required. During attachment PMNs become activated and release a variety of substances, including toxic oxygen species such as superoxide and hydrogen peroxide. In the in vitro system used in this study no injury occurred to the endothelial cells, as measured by 51Cr release, when activated PMNs were added with ceruloplasmin. The data show that ceruloplasmin reduced, in a dose dependent manner, the levels of superoxide produced by the activated PMNs, further supporting ceruloplasmin's previously reported role as a scavenger of superoxide. Ceruloplasmin also reduced the levels of superoxide when activated PMNs were in contact with endothelial cells. Although ceruloplasmin interfered with the copper-dependent scavenger enzyme, superoxide dismutase (SOD), in a cell-free system, ceruloplasmin had no effect on SOD in intact endothelial cells. These results suggest that ceruloplasmin may act as an anti-inflammatory agent by reducing the number of PMNs attaching to endothelium and by acting as an extracellular scavenger of superoxide. PMID- 2552812 TI - Cell deletion by apoptosis during regression of renal hyperplasia. AB - Regression of renal hyperplasia after withdrawal of the mitogenic stimulus induced by a single injection of lead nitrate was studied in male Wistar rats. Lead nitrate administration (10 mumol/100 g body weight) resulted in a ninefold increase in the incorporation of labeled thymidine into renal DNA and in an enhancement in the mitotic index; these changes were accompanied by an increase in the organ weight and DNA content that reached a maximum at 2 days. Regression of the renal hyperplasia was observed as early as 3 days after treatment and was completed within 2 weeks. Although lytic necrosis was not responsible for cell loss, the elimination of the excess renal cells took the form of apoptosis. This distinctive mode of cell death, which has been implicated in the involution of hyperplasia in other tissues and organs, was characterized by the occurrence of intracellular and extracellular membrane-bounded eosinophilic globules that often contained nuclear fragments. It affected mainly cells of the proximal tubules, and it was not detected once the kidney had regressed to its original mass. These results support the hypothesis that apoptosis is involved in the regulation of organ size. PMID- 2552813 TI - Primary polyoma virus-induced murine thymic epithelial tumors. A tumor model of thymus physiology. AB - Thymic tumors were induced in C3'/Bittner mice by neonatal inoculation with polyoma virus. The objective of this study was to identify the phenotypes of the cells within the tumors and to attempt to determine the origin of the neoplastic cell population(s). At the ultrastructural level, the neoplastic cells resembled normal thymic epithelium with tonofilaments and desmosomes. Immunoperoxidase staining demonstrated the presence of cytokeratin, Iak, -beta 2-microglobulin, asialo-GM1, the thymic cortical epithelial marker ER-TR4, and the medullary epithelial marker ER-TR5. Islands of normal cortical thymocytes supported by residual normal cortical epithelium and acid phosphatase-positive cortical macrophages were interspersed in the tumors. Residual islands of normal medullary architecture with nonspecific esterase-positive IDCs were rarely identified in tumors. Most lymphocytes in the tumors were normal immature cortical thymocytes with the phenotype Tdt+, PNA+, Thy 1.2bright, Ly-1dull, H-2Kkdull, ThB+, J11d+, and Lyt-2+L3T4+. Lymphocytes in the tumors were steroid-sensitive like normal thymocytes. The proportions of Lyt-2+L3T4- and Lyt-2-L3T4+ cells were generally larger in the tumors than in normal thymus and reflected the higher frequency of lymphocytes in the tumors capable of proliferating in vitro in response to Con A plus IL-2. The data were consistent with the hypothesis that the neoplasia originates from thymic epithelium that is interspersed with normal, developing thymic lymphocytes. PMID- 2552814 TI - Human papillomavirus detection in paraffin-embedded cervical carcinomas and metastases of the carcinomas by the polymerase chain reaction. AB - The polymerase chain reaction (PCR) is used for human papillomavirus (HPV) detection in paraffin-embedded tissue. The specificity of the reaction is unaffected by the method of fixation used before embedding into paraffin. Five HPV 16, 18, 31, and 33 DNA in situ hybridization (DISH)-negative cervical carcinomas were subjected to the PCR. In two patients, HPV 16 DNA could be detected in the cervical squamous cell carcinomas and also in their lymph node metastases. One patient with an adeno-carcinoma of the cervix was found positive for HPV-18. A lymph node of this patient was HPV 18 positive as well. In the tumors of the remaining two patients, no HPV 16, 18, or 33 DNA was detected by the PCR. Both negative patients had cervical squamous cell carcinomas. One had a bladder metastasis, whereas the other had a lymph node metastasis and an additional distant metastasis in the lung. HPV DNA positivity in cervical carcinomas correlated with HPV prevalence in the metastases. This relationship can be of use for diagnostic purposes in the pathologic analysis of metastases. PMID- 2552815 TI - Auditory acuity in adrenocorticoid insufficiency. AB - Studies in man have proposed that individuals with adrenocorticoid insufficiency show a heightened auditory detection threshold. This hyperacuity was shown to be specific for deficits in carbohydrate-active steroids, with threshold levels improved at least 13 dB between 1,000 Hz and 4,000 Hz. These reversible changes in auditory detection have been related to specific influences of glucocorticoids on the metabolism of neural tissue. The present study was designed to examine the hypothesis that glucocorticoid deficiencies cause decreased electrocochleographically measured auditory thresholds with interference in the central conduction of these impulses. A rat model was employed in an attempt to demonstrate the mechanism for a state of increased neural excitability in the absence of carbohydrate-active steroid influence. Electrocochleographic and auditory brain-stem evoked response threshold and latency determinations were made in eight experimental rats. Adrenalectomies were then performed in all animals with postoperative electrocochleographic and auditory brain stem responses being made at 3, 7, and 21 days. Threshold recordings for all experimental animals showed no significant changes following the cessation of glucocorticoid production. Latency measurements revealed impaired central neural transmission of the first and second order neurons. The possible mechanisms for these findings are discussed. PMID- 2552816 TI - Insulin and IGF I receptor-mediated Na+ transport in toad urinary bladders. AB - We compared the concentration dependence of insulin- and insulin-like growth factor I (IGF I)-stimulated Na+ transport with ligand-receptor affinities in the urinary bladder of the toad Bufo marinus. Threshold, half-maximal, and maximal natriferic concentrations of both peptides were approximately 0.1, 1, and 10 nM, respectively. Amiloride, but not ethyl isopropyl amiloride, (10(-5) M), abolished Na+ transport. Maximal responses to either peptide rendered the tissue insensitive to challenge with the other. Separate insulin and IGF I receptors were identified by equilibrium binding and polyacrylamide gel electrophoresis of cross-linked ligand-receptor complexes. For both peptides, half-maximal binding occurred at 3-10 nM; crossover binding to the other receptor occurred with 10- and 100-fold lower affinity. Thus, in this model "high-resistance" renal epithelium, 1) ligand binding to specific insulin and IGF I receptors stimulates transcellular Na+ flux, 2) the natriferic effects of insulin and IGF I apparently depend on activation of apical Na+ channels rather than Na+-H+ antiporters, and 3) the natriferic pathways activated by insulin and IGF I appear to converge subsequent to ligand-receptor binding but before the final transport ("effector") step(s). PMID- 2552817 TI - Parathyroid hormone-induced translocation of Na-H antiporters in rat proximal tubules. AB - Parathyroid hormone (PTH) is believed to inhibit bicarbonate reabsorption by inhibiting Na-H antiport activity in proximal tubular brush-border membranes. The sequence of events triggered by PTH was investigated in a crude preparation of proximal tubules obtained by mechanical disruption and filtration through nylon mesh filters. Tubule samples were subjected to analytical subcellular fractionation after 2-, 5-, and 30-min treatments with 1 IU/ml PTH. These PTH treatment intervals caused 54, 63, and 68% decreases in the Na-H antiport activity of a population of brush-border membrane vesicles that was resolved from a PTH-unresponsive brush-border population by density-gradient centrifugation. The rapid loss of Na-H antiport activity from the responsive population was accompanied by a transient increase in the Na-H antiport activity of a region of the density gradient, designated density window III, which was shown to contain two distinct membrane populations; these populations were both enriched in acid phosphatase activity, and one of them was also an important locus of galactosyltransferase activity. The increase in the Na-H antiport activity of window III accounted for 52% of the activity lost from the PTH-responsive population after 2 min, and for 43% of the activity lost after 5 min, but it was completely abolished after 25 more minutes in the presence of PTH. These observations suggest that PTH triggers a rapid translocation of Na-H antiporters from the microvillus membrane to a distinct membrane domain, where they are subsequently inactivated. PMID- 2552818 TI - D 600 block of L-type Ca2+ channel in vascular smooth muscle cells: comparison with permanently charged derivative, D 890. AB - It has been reported that D 600 blocks the high-threshold Ca2+ channel (L-type) from the outside in isolated vascular and ileal smooth muscle cells of the rabbit (Y. Ohya, K. Terada, K. Kitamura, and H. Kuriyama. Pfluegers Arch. 408: 80-82, 1987). We have reinvestigated this hypothesis by comparing the effects of external and internal applications of D 600 and the permanently charged quaternary derivative D 890 on the whole cell Ca2+ current (Ica) recorded in vascular smooth muscle cells isolated from the rabbit portal vein. At low frequencies of stimulation (0.05 Hz), externally applied D 600 inhibited Ica in a dose-dependent fashion, with a complete block occurring at 10(-4) M. D 600 was approximately 1,000 times more potent than D 890 for causing inhibition of Ica using this protocol. During a train of stimulations at 0.5 Hz, D 600 (10(-6) M) produced a minor additional frequency-dependent block of Ica, as shown in other preparations. During superfusion with D 890 (10(-4) M), a similar protocol produced little if any decline in the amplitude of Ica. No evidence of block could be detected during intracellular dialysis of D 600 (10(-4) M). At the same concentration, intracellular application of D 890 produced a slow block of Ica. To test whether D 600 could be effectively dialysed using a patch micropipette, similar experiments were performed in cardiac ventricular myocytes. In this preparation, intracellular dialysis of D 600 induced a rapid inhibition of the Ica.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552819 TI - Enhanced erythropoietin secretion in hepatoblastoma cells in response to hypoxia. AB - Erythropoietin (Ep) levels in spent culture media of a Hep G2 human hepatoblastoma cell line were measured by radioimmunoassay (RIA), fetal mouse liver erythroid colony formation (FMLC), and the exhypoxic polycythemic mouse assay (EHPCMA). The Hep G2 cells at high density produced approximately 700 mU/ml Ep when measured with the RIA. On the other hand, the Ep levels when assayed in EHPCMA and FMLC were 50 and 2,600 mU/ml, respectively. The bioactivity in FMLC was completely neutralized by an antibody to purified human recombinant Ep, indicating that the erythropoietic activity in the Hep G2 spent culture medium was immunologically equivalent to Ep. Ep levels in the medium from low-density Hep G2 cells in 5% O2 and 1% O2 were 2.5- and 4-fold greater, respectively, than that of 20% O2. In contrast, hyperoxia (40% O2) significantly inhibited Ep production. A significant increase in Ep secretion was also observed when the cells were incubated with cobaltous chloride (2 X 10(-6) -2.5 X 10(-4) M). Tunicamycin (0.5 micrograms/ml), which inhibits N-linked glycosylation, significantly reduced the enhancement of Ep secretion induced by hypoxia (1% O2) without affecting cell growth. Forskolin and cholera toxin, each of which increased the levels of cyclic AMP in the Hep G2 cells by 40-fold, produced a significant (P less than 0.05) further increase in Ep secretion in the presence of hypoxia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552820 TI - Malignant hyperthermia: slow sodium current in cultured human muscle cells. AB - Voltage-activated ion currents were measured in cultured skeletal muscle myoballs. Cultures were generated from biopsies from patients referred for diagnosis of susceptibility to malignant hyperthermia (MH); diagnosis of susceptibility (MH+) or nonsusceptibility (MH-) was made on the basis of in vitro halothane-induced contracture of a separate piece of biopsy. Measurements of ion currents were made at room temperature in the absence of anesthetic agents, using tight-seal whole-cell recording. Fast transient Na+ currents and delayed outward K+ currents were similar in magnitude and kinetics in cells from MH+ and MH- patients. An additional slowly inactivating inward current component was commonly observed in cells from MH+ patients. This current was blockable by tetrodotoxin and was carried by Na+ but not by Ba2+. The component was less frequently observed and was of a lower magnitude in cells from MH- patients. The increased magnitude of the slow inward current observed in cultured muscle cells from MH+ patients may be a manifestation of the lesion that causes MH. PMID- 2552821 TI - Downregulation of beta-adrenergic receptor-mediated function during sodium restriction in humans. AB - Mononuclear leukocyte (MNL) beta 2-adrenergic receptor (beta 2-AR) binding and its linked adenylate cyclase sensitivity to isoproterenol were measured in nine healthy humans prior to and after 7 days of dietary sodium restriction to determine whether chronic physiological increases in plasma norepinephrine (NE) are associated with the downregulation of beta-AR-mediated function. Sodium restriction resulted in an increase in the plasma NE concentration (P less than 0.02) and decreases in MNL beta 2-AR density (P less than 0.001), affinity for antagonist (P less than 0.001), and adenylate cyclase sensitivity to isoproterenol (ANOVA, P less than 0.01). To determine whether this downregulation of MNL beta 2-AR-mediated function is related to the increased plasma NE concentration or to increased extravascular NE release, NE kinetics was assessed using compartmental analysis in each subject prior to and after sodium restriction. Sodium restriction caused a decrease in the plasma NE metabolic clearance rate (P less than 0.005) and in the volume of distribution of NE in the intravascular compartment (P less than 0.005), whereas the extravascular NE release rate was unchanged. Our data suggest that the downregulation of MNL beta 2-AR-mediated function in humans during dietary sodium restriction is a response to the increase in plasma NE. PMID- 2552822 TI - Demonstration and modification of intervillous pH profiles in rat small intestine in vitro. AB - The intervillous pH profiles along the crypt villus axis in different regions of the rat small intestine were measured in vitro by using pH-sensitive liquid ion exchanger microelectrodes. A characteristic pH profile was observed in the duodenum and jejunum. A region of low pH was detected in the upper parts of the villi (pH 6.65 +/- 0.06 to 6.85 +/- 0.07), whereas pH at the villus base was always higher. In the ileum no gradient was observed (pH 7.26 +/- 0.05 to 7.31 +/ 0.05). Preincubation of the tissue in situ with 10 mM theophylline for 1 h caused an increase in the villus base pH in the jejunum (pH 7.24 +/- 0.04) and ileum (7.44 +/- 0.04) followed by a subsequent increase of the pH in the upper part of the villi. These results indicate that the low pH in the upper intervillous space may be related to H+ secretion occurring from the mature enterocytes, whereas the crypt cells may secrete a rather neutral or slightly alkaline fluid. Alkaline secretion from the crypts may be increased by theophylline, which changes the levels of cyclic nucleotides in the mucosa. PMID- 2552823 TI - Angiotensin receptors and angiotensin I-converting enzyme in rat intestine. AB - The purpose of this study was to map the distribution of angiotensin II (ANG II) receptors and ANG I-converting enzyme (ACE) in rat intestine. ANG II binding sites were visualized by in vitro autoradiography using iodinated [Sar1, Ile8]ANG II. The distribution of ACE was mapped using an iodinated derivative of lisinopril. Male Sprague-Dawley rats were killed and the interior of the whole intestine washed with ice-cold saline. Segments of duodenum, jejunum, ileum, and colon were quickly frozen in a mixture of isopentane and dry ice. Twenty-micron frozen sections were thaw-mounted onto gelatin-coated slides, incubated with either ligand, and exposed to X-ray film. After exposure and subsequent development, the films were quantitated by computerized densitometry. ANG II receptors were most dense in the colon, followed by the ileum, duodenum, and jejunum. Within each segment of intestine, specific ANG II binding sites were localized exclusively to the muscularis. In contrast, ACE was present in both the mucosa and the muscularis. The colocalization of ANG II receptors and ACE may suggest a role for locally generated ANG II in the control of intestinal function. The luminal orientation of ACE in the mucosa of the small intestine may suggest that at this site ACE serves primarily to hydrolyze dietary peptides. PMID- 2552824 TI - Gastric H+-K+-ATPase in situ: evidence for compartmentalization. AB - Gastric glands, isolated from rabbit, were permeabilized with digitonin to permit measurement of H+-K+-adenosinetriphosphatase (ATPase) activity and proton transport in situ. Measurement of proton gradient formation using acridine orange fluorescence showed two phases of ATP-driven proton accumulation; one phase occurs spontaneously in KCl medium and one phase requires the K+ ionophore valinomycin. Valinomycin was found to increase H+-K+-ATPase activity, indicating that the second phase is because of increased proton transport rather than a decrease in proton leak rate. The acid-activated, irreversible inhibitor, omeprazole, was used to selectively eliminate the H+-K+-ATPase molecules associated with the spontaneous component of proton transport. After omeprazole treatment a residual, valinomycin-dependent component of proton transport could be demonstrated. These results are interpreted as evidence for two compartments of H+-K+-ATPase, separated by a barrier that prevents K+ diffusion and pH equilibration. The two compartments may be separated also on the basis of anion selectivity. The spontaneously active compartment was found to be functional with various anions, including sulfate and isethionate, whereas the valinomycin dependent component is highly selective for chloride. The proportion of H+-K+ ATPase that exists in each compartment was quantitated by measuring the fraction of total ATPase activity that could be inhibited by omeprazole in the absence and presence of valinomycin. For glands that were preconditioned with cimetidine, approximately 30% of the inhibitable enzyme was found associated with the spontaneous compartment, and this fraction increased to approximately 70% with histamine preconditioning.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552825 TI - Vasoactive intestinal peptide and its receptors in fetuses with cystic fibrosis. AB - Fetuses were investigated to establish whether vasoactive intestinal peptide (VIP) and its receptors are involved in the basic biochemical defect causing cystic fibrosis (CF). The intestine was used as a target for the disease and the liver as control. The immunoreactive and biologically active VIP contents of the colon and lower part of the small intestine were 1.5-2.5 times higher in CF fetuses than in controls. In control and CF intestinal mucosa, there was no change in the Scatchard parameters of the 125I-labeled VIP binding sites (Kd = 4.7-6.1 X 10(-11) M; Bmax = 268-280 fmol/mg protein for the high-affinity sites), in the two molecular components constituting the cross-linked 125I-VIP binding (Mr = 66,000 and 30,000), or in the pharmacological properties and functional characteristics of the VIP receptors activating the G proteins-adenylate cyclase system (Ka = 0.7 X 10(-9) M VIP). Similar results were obtained in liver. These findings suggest that neither VIP nor its receptors are involved in CF intestine. The possible involvement of other effectors related to the VIP pathway in CF intestine, including the release of VIP and adenosine 3',5'-cyclic monophosphate signal-transduction cascade, are presented. PMID- 2552826 TI - Altered smooth muscle contraction and sodium pump activity in the inflamed rat intestine. AB - We examined changes in membrane function underlying the increased contractility of jejunal longitudinal muscle to carbachol in rats infected 6 days previously with Trichinella spiralis. Muscarinic receptor characteristics were examined in particulate fractions using [N-methyl-3H]scopolamine (NMS). There was a significant reduction in the total number of binding sites on muscle from infected rats, but the affinity for NMS was unchanged. Similarly, in competition studies, the binding of carbachol to high or low affinity sites was not significantly different in tissue from control or infected rats. However, we observed an 89% suppression of the activity of K+ -stimulated ouabain-sensitive p nitrophenylphosphatase (pNPPase), an enzyme marker for the Na+ -K+ pump, in plasma membranes from infected compared with control rats. Similar results were obtained in 86Rb uptake studies. In contractility studies, evidence for the electrogenicity of the Na+ -K+ pump was obtained by demonstrating that pump activation by K+, Rb+, or Cs+ was associated with tissue relaxation with a rank order of potency that was identical to that for stimulation of pNPPase activity by these ions. Conversely, pump inhibition by vanadate increased tone and abolished phasic contractions in muscle from control or infected rats. This was accompanied by an increased response to carbachol in muscle from control but not infected rats. In addition, pump inhibition by removing extracellular K increased tone in control tissue but decreased tone in muscle from T. spiralis-infected rats, presumably because of preexisting pump suppression. These results are consistent with the hypothesis that suppression of electrogenic Na-pump activity contributes to the increased contractility of jejunal muscle in rats infected with T. spiralis. PMID- 2552827 TI - Na+-H+ antiporter of rat colonic basolateral membrane vesicles. AB - The present experiments were conducted, using acridine orange and 22Na uptake techniques, to demonstrate the presence of an electroneutral Na+-H+ exchange process in rat colonic basolateral membrane vesicles. Results consistent with the existence of a distinct Na+-H+ antiporter in these vesicles include the following: 1) an outwardly directed Na+ gradient stimulated proton influx (Na+in, 100 mM; pHin 7.5/pHout 7.5) and an inwardly directed sodium gradient (Na+out, 5 50 mM; pHin 6.0/pHout 7.5) stimulated proton efflux; 2) sodium-stimulated proton influx was minimally decreased (approximately 10-25%) under voltage clamp conditions (addition of valinomycin in the presence of K+ on both sides of vesicles), indicating that Na+ for H+ exchange in these vesicles could not be explained solely on the basis of a membrane potential; 3) an outwardly directed proton gradient (pHin 5.5/pHout 7.5) stimulated 22Na uptake into these vesicles and a threefold "over-shoot" was observed; 4) 22Na uptake and sodium-stimulated proton efflux were saturable with a Km for Na+ of 5.8 +/- 0.9 and 7.0 +/- 0.3 mM, respectively; 5) amiloride (1 mM) significantly inhibited both sodium-stimulated proton efflux (approximately 69%) and 22Na uptake (approximately 89%), but other transport inhibitors (acetazolamide, 4-acetamido-4'isothiocyanostilbene-2,2' disulfonic acid, and bumetanide) had no effect on 22Na uptake; 6) N methylglucamine+ (a nonpermeant cation) did not affect pH gradient-stimulated 22Na uptake, whereas Li+ inhibited Na+ uptake; 7) an inwardly directed Li+ gradient stimulated proton efflux and the Km for Li+ was 12.5 +/- 1.5 mM. These findings establish the existence of an electroneutral Na+-H+ exchange mechanism in rat colonic basolateral membrane vesicles. PMID- 2552828 TI - Sublethal oxidant injury inhibits signal transduction in rat type II pneumocytes. AB - The production of reduced forms of O2 in oxidant injury can lead to lipid peroxidation, which would pose a threat to cell membrane integrity and therefore to signal transduction pathways located in the membrane. We studied the effects of oxidant injury with t-butyl hydroperoxide (tBOOH) on signal transduction in rat type II pneumocytes in culture. Exposure of 1 x 10(6) type II pneumocytes to tBOOH concentrations less than or equal to 100 microM did not cause significant release of lactate dehydrogenase and was therefore termed sublethal. Exposure to sublethal concentrations of tBOOH caused a dose-dependent inhibition of surfactant secretion stimulated both by terbutaline (10(-4)M) and by extracellular ATP (10(-5)M). Adenosine 3',5'-cyclic monophosphate production in response both to terbutaline and to extracellular ATP was also inhibited by exposure to 100 microM tBOOH. In addition, exposure to 100 microM tBOOH inhibited inositol phosphate formation in response to extracellular ATP. We conclude that sublethal oxidant injury inhibits not only receptor-mediated agonist stimulation of surfactant secretion but also receptor-mediated agonist stimulation of second messenger formation in type II pneumocytes in culture. Such inhibition may be important in the pathogenesis of both the adult and neonatal forms of respiratory distress syndrome, in which alveolar surfactant deficiency may be exacerbated by oxidant injury. PMID- 2552829 TI - Histamine and inositol phosphate accumulation in endothelium: cAMP and a G protein. AB - Histamine increases microvascular permeability through a calcium-dependent process, and histamine occupancy of the H1-receptor increases calcium in cultured endothelial cells. Agents that increase adenosine 3',5'-cyclic monophosphate (cAMP) in endothelial cells prevent the in vivo increase in microvascular permeability that follows histamine exposure. In the current experiments, histamine occupancy of the H1-receptor increased the flux of albumin across monolayers of cultured human umbilical vein endothelial cells (HUVEC). This was prevented by pretreating the cells with theophylline, forskolin, and 8-bromo-cAMP (BrcAMP), which also decreased the flux of albumin across control monolayers. Exposing the cells to histamine increased inositol phosphate accumulation in the cells, and this was prevented by the H1-antagonist pyrilamine but not by theophylline, forskolin, and BrcAMP. Exposing the cells to histamine increased intracellular calcium measured with fura-2. The increase in cell calcium was prevented by pyrilamine but not by pretreatment with theophylline, forskolin, and BrcAMP. When endogenous cell GTP was depleted by permeabilizing the membranes of the endothelial cells with Staphylococcus aureus alpha-toxin, histamine stimulated inositol phosphate accumulation was enhanced with addition of GTP but not with addition of GDP to the buffer. Addition of GTP alone to the buffer did not increase inositol phosphate accumulation in alpha-toxin-treated cells. Histamine stimulates inositol phosphate accumulation in HUVEC via a G protein. Inhibition of the edemagenic effects of histamine by cAMP does not occur by interrupting this signal transduction pathway between the binding of histamine to its receptor and the increase in intracellular calcium. PMID- 2552830 TI - Selective expression of an amiloride-inhibitable Na+ conductance from mRNA of respiratory epithelium in Xenopus laevis oocytes. AB - Poly(A)+RNA was prepared from primary cultures of human nasal polyp epithelia and from native bovine tracheal epithelia. Six to fifty nanograms mRNA were injected into prophase-arrested immature Xenopus laevis oocytes. One to four days later the oocytes were probed with electrophysiological techniques for induction of novel ion conductances. Oocytes injected with mRNA had lower membrane potentials (Vm) and resistances (Rm) than controls. By use of step changes in extracellular Na+ concentration and applying amiloride, a Na+ conductance could be identified in mRNA-injected oocytes, which was inhibited by submicromolar concentrations of amiloride with the same kinetics (Ki = 1.3 X 10(-7) mol/l) as in the original tissue. After complete inhibition of this conductance by 10(-5) mol/l amiloride, Vm and Rm approached the respective values of controls. The data indicate that oocytes express functional epithelial Na+ channels but apparently no other epithelial ion conductances from injected mRNA of respiratory epithelium. PMID- 2552831 TI - Atrial natriuretic factor binding sites in experimental congestive heart failure. AB - A quantitative in vitro autoradiographic study was performed on the aorta, renal glomeruli, and adrenal cortex of cardiomyopathic hamsters in various stages of heart failure and correlated, in some instances, with in vivo autoradiography. The results indicate virtually no correlation between the degree of congestive heart failure and the density of 125I-labeled atrial natriuretic factor [(Ser99, Tyr126)ANF] binding sites (Bmax) in the tissues examined. Whereas the Bmax was increased in the thoracic aorta in moderate and severe heart failure, there were no significant changes in the zona glomerulosa. The renal glomeruli Bmax was lower in mild and moderate heart failure compared with control and severe heart failure. The proportion of ANF B- and C-receptors was also evaluated in sections of the aorta, adrenal, and kidney of control and cardiomyopathic hamsters with severe heart failure. (Arg102, Cys121)ANF [des-(Gln113, Ser114, Gly115, Leu116, Gly117) NH2] (C-ANF) at 10(-6) M displaced approximately 505 of (Ser99, Tyr126)125I-ANF bound in the aorta and renal glomeruli and approximately 20% in the adrenal zona glomerulosa in both series of animals. These results suggest that ANF may exert a buffering effect on the vasoconstriction of heart failure and to a certain extent may inhibit aldosterone secretion. The impairment of renal sodium excretion does not appear to be related to glomerular ANF binding sites at any stage of the disease. PMID- 2552832 TI - Divergent effects of KCl-induced depolarization on afferent and efferent arterioles. AB - We have previously proposed that renal microvessels exhibit a unique regional heterogeneity. Studies with calcium channel agonists and antagonists suggest that potential-dependent calcium channels may play a more prominent role in the activation of the afferent arteriole than the efferent arteriole. Because KCl induced depolarization elicits vasoconstriction exclusively by the activation of potential-dependent calcium channels, we tested this postulate directly by ascertaining the vasoconstrictor effects of KCl and countervailing effects of a calcium channel blocker on the afferent and efferent arteriole of isolated perfused hydronephrotic kidneys. Increasing media potassium concentration from 5 to 30 mM resulted in a marked renal vasoconstriction decreasing renal perfusate flow by 61 +/- 4%. An examination of the microvascular response to KCl revealed a predominant response of the afferent arteriole. Thus afferent arteriolar diameter decreased by 38 +/- 6% (i.e., from 20.7 +/- 1.5 to 13.0 +/- 1.8 microns, P less than 0.005), whereas efferent arteriolar diameter decreased by only 12 +/- 4% (i.e., from 15.8 +/- 1.6 to 13.8 +/- 1.4 microns, P = 0.05). Nifedipine completely returned afferent arteriolar diameter to control levels with a mean effective dose of 41 +/- 2 nM. These findings indicate that the afferent arteriole is more responsive to depolarization-induced vasoconstrictor stimuli than is the efferent arteriole and suggest a greater prevalence of potential dependent calcium channels in this vessel. PMID- 2552833 TI - Direct Na+-K+ pump stimulation by K+ in cortical collecting tubules: a mechanism for early renal K+ adaptation. AB - To evaluate the mechanism of increased Na+-K+ pump turnover rate that characterizes the early cortical collecting tubule (CCT) response to K+ loading [Y. Fujii, S. K. Mujais, and A. I. Katz. Am. J. Physiol. 256 (Renal Fluid Electrolyte Physiol. 25): F279-F284, 1989.], we measured ouabain-sensitive 86Rb+ uptake in microdissected rat CCT exposed acutely to elevated ambient K+ in vivo and in vitro. Tubules preincubated in 10 mM K+ had higher 86Rb+ uptake than when preincubated in 5 mM K+ (25.9 +/- 1.2 vs. 18.9 +/- 0.7 pmol.mm-1.min-1, P less than 0.001). KCl infusion (5 mumol.100 g-1.min-1 x 60 min) increased 86Rb+ uptake from 19.2 +/- 1.0 to 31.2 +/- 1.4 pmol.mm-1.min-1, P less than 0.001; the increment was preserved in tubules subsequently treated with monensin or nystatin in vitro, suggesting that pump stimulation was not mediated by increased cell Na+. This conclusion was confirmed in separate experiments in which the effect of K+ on 86Rb+ uptake was not altered by concurrent preincubation with amiloride. Studies with CCT from isolated perfused kidneys and from adrenalectomized animals revealed that stimulation of 86Rb+ uptake by a K+ load occurs rapidly (less than or equal to 5 min) and is independent of hormonal factors. Increased external K+ produces a rapid rise in K+-transporting capacity (turnover rate) of the Na+-K+ pump in CCT. This phenomenon probably represents a direct effect on K+ on the pump and is an important component of the early renal response to increased K+ secretory load. PMID- 2552834 TI - Adaptation of rabbit renal cortical Na+-H+ exchange activity in chronic hypocapnia. AB - We have examined the activity and kinetic characteristics of the Na+-H+ exchanger in renal cortical brush-border membrane vesicles (BBMV) prepared from rabbits adapted to chronic hypocapnia in order to address whether this transporter might contribute to the suppressed proximal bicarbonate reabsorption characteristic of this disorder. Chronic hypocapnia was induced by exposing animals to 9% O2 for a 5-day period. In comparison with paired, contemporaneous controls, an average delta PaCO2 of 13 mmHg and an average delta [HCO3-] of 7.3 meq/l were obtained. Chronic hypocapnia led to a significant suppression of the 22Na+ uptake by BBMV; at the 3-s mark, a 30% suppression was observed (chronic hypocapnia, 4.05 +/- 0.43 nmol/mg protein; control, 5.72 +/- 0.39 nmol/mg protein) (P less than 0.01). A significant decrease in the Vmax of the antiporter was noted (chronic hypocapnia, 622.7 +/- 86.8 nmol.mg protein-1.min-1; control 857.5 +/- 64.8 nmol.mg protein-1.min-1) (P less than 0.01), whereas the Km for sodium remained unaltered. The specificity of this adaptation was supported by showing that Na+ dependent uptake of D-[3H]glucose by BBMV was not significantly different between chronic hypocapnia and control. Chronic normocapnic hypoxemia left Na+-H+ exchange activity undisturbed. We conclude that the observed change in the BBMV Na+-H+ antiporter might be responsible, at least in part, for the suppressed renal bicarbonate reabsorption characteristic of chronic hypocapnia and that a consequence of the hypocapnic state itself mediates this adaptation. PMID- 2552835 TI - Expression and regulation of ANP receptor subtypes in rat renal glomeruli and papillae. AB - The expression and regulation of atrial natriuretic peptide (ANP) receptor subtypes were examined in rat renal glomeruli and papillae. In glomeruli, approximately two-thirds of ANP binding sites represented guanylate cyclase uncoupled ANP clearance receptors (ANPc) with a molecular mass of 64 kDa under reducing conditions. The remainder of glomerular ANP binding sites represented guanylate cyclase-coupled ANP receptors (ANPGC) with a molecular mass of 130 kDa. In rat papillae, only the 130-kDa ANPGC was expressed. In rats adapted to a low salt diet, saline loading or acute ANP infusion resulted in a decrease in ANPC density, a difference that was not detected when glomeruli were first acidwashed to remove endogenous ANP, indicating that apparent regulation of ANPC reflected prior occupancy by endogenous ANP. Densities of glomerular ANPC and ANPGC were similar in spontaneously hypertensive rats (SHR) compared with those of the Wistar-Kyoto (WKY) controls. However, elimination of prior receptor occupancy revealed a significantly greater expression of glomerular ANPC in SHR compared with WKY rats, without significant differences in the density of the glomerular or papillary ANPGC subtype. The failure of the ANPGC subtype to be regulated may account for our previously reported findings that dietary salt intake does not affect glomerular ANP-stimulated guanosine-3',5'-cyclic monophosphate accumulation despite apparent regulation of ANP receptor density. Whether the increased expression of the ANPC subtype in SHR represents a primary defect or results from induction of ANP clearance receptor expression remains to be determined. PMID- 2552836 TI - Decrease in ureagenesis by partial hepatectomy does not influence acid-base balance. AB - It has been suggested that urea synthesis participates directly in body pH homeostasis by removal of bicarbonate. To elucidate this hypothesis sodium bicarbonate or sodium chloride was infused (11.5 mumol/min) for 90 min into control rats and into rats that had undergone an 85% hepatectomy immediately before starting the infusion. Urea synthesis rate was 2.6 +/- 0.3 mumol/min (mean +/- SE) in controls, and was significantly (P less than 0.01) reduced to 1.0 +/- 0.2 mumol/min in partially hepatectomized rats. At the start of bicarbonate infusion, pH was 7.38 and 7.34 in control and partially hepatectomized rats, respectively, and at the end of infusion, pH was 7.56 and 7.51. Standard bicarbonate at start of bicarbonate infusion was 21.9 and 21.3 mM in controls and partially hepatectomized, respectively, and it increased to 32.7 and 29.9 mM at end of infusion. In saline-infused rats a slight decrease of approximately 0.05 pH units was observed during the experiment, but again no difference emerged between control and partially hepatectomized rats. It is concluded that a major role of the liver in the regulation of acid-base balance is unlikely. PMID- 2552837 TI - Adenosine receptor coupling to adenylate cyclase of rat ventricular myocyte membranes. AB - The effects of adenosine analogues on beta-adrenergic receptor and receptor independent elicited increases in adenylate cyclase activity were investigated using membranes obtained from primary cultures of adult rat ventricular myocytes. Phenylisopropyladenosine, an A1-receptor agonist, at concentrations of 0.1, 1.0, and 10 microM, maximally inhibited isoproterenol-stimulated adenylate cyclase activity by 35, 55, and 41%, respectively. The inhibition by phenylisopropyladenosine was antagonized by 10 microM theophylline. One micromolar phenylisopropyladenosine was much less effective at attenuating forskolin-stimulated activity, such that the maximum inhibition was 26%. Phenylisopropyladenosine had no effect on adenylate cyclase stimulation by 5' guanylylimidodiphosphate. Phenylaminoadenosine, an A2 agonist, at 10 microM or greater stimulated adenylate cyclase activity. This effect was not significantly inhibited by theophylline or 0.1 microM 1,3-dipropyl-8-cyclopentylxanthine (DPCPX), which antagonized phenylisopropyladenosine inhibition of isoproterenol stimulated adenylate cyclase activity. Additionally, N-ethylcarboxamidoadenosine, a nonselective adenosine-receptor agonist, had no effect on adenylate cyclase activity in the absence of DPCPX but stimulated adenylate cyclase activity in the presence of DPCPX. These results indicate that both A1 and A2 receptors exist on the ventricular myocyte sarcolemma. More importantly, the findings suggest that adenosine inhibition of catecholamine-stimulated adenylate cyclase activity is primarily due to an alteration in beta-adrenergic receptor-mediated transduction and perhaps in part by a direct inhibition of the catalytic component. PMID- 2552838 TI - Alpha-adrenergic effects on relative refractory period in Purkinje system of intact canine left ventricles. AB - To investigate the electrophysiological effects of alpha-agonists, we studied 23 chloralose-anesthetized sinoaortic and vagotomized dogs, measuring epicardial and endocardial effective refractory period and relative refractory period of the Purkinje system during graded infusions of norepinephrine and phenylephrine. In group 1, epicardial and endocardial effective refractory periods shortened equivalently with norepinephrine. These effects were blocked with metoprolol. In group 2, epicardial and endocardial refractory periods did not prolong with phenylephrine despite addition of metoprolol. In group 3, phenylephrine, after the addition of metoprolol, prolonged the relative refractory period of Purkinje only at the highest phenylephrine dose of 50 micrograms.kg-1.min-1. In group 4, these latter effects were not produced by raising pressure with descending aortic occlusion. However, the effects of phenylephrine were blocked with prazosin (1.0 mg/kg). Taken together, these data demonstrate that alpha-adrenergic influences prolong Purkinje but not muscle refractory period in the intact canine ventricle. The high dose of phenylephrine and resulting hypertension suggest that this may be a pharmacological and not a physiological response. PMID- 2552839 TI - Phorbol amplification of serotonin-induced arterial contractions is endothelium dependent. AB - This study determined the effect of phorbol pretreatment on serotonin [5 hydroxytryptamine (5-HT)]-induced contractions of rabbit abdominal aorta. Rings of aorta pretreated with 12-O-tetradecanoylphorbol 13-acetate (TPA, 10(-7) M) contracted more strongly (P less than 0.01) than rings pretreated with the vehicle dimethyl sulfoxide (0.0007%), but only at low 5-HT concentrations (10(-8) to 10(-7) M). A 10-min exposure to TPA (10(-7) M) increased (P less than 0.01) 5 HT contractions (10(-7) M) from 23.7 +/- 2.6 to 42.1 +/- 4.5%. This amplification required an intact endothelium and the use of a biologically active phorbol. Pretreatment with TPA (10(-7) M) reduced (P less than 0.01) acetylcholine-induced relaxations from 56.6 +/- 6.6 to 20.0 +/- 13.8%. Changes in guanosine 3',5' cyclic monophosphate-mediated relaxation are not involved in this decrease, since relaxations to sodium nitroprusside (10(-9) to 10(-8) M) were not altered by TPA pretreatment. These results suggest that activation of protein kinase C in endothelial cells may produce arterial supersensitivity to 5-HT by decreasing the release of endothelium-derived relaxing factor(s) and/or by increasing the release of endothelium-derived contracting factor(s). PMID- 2552840 TI - Similar responsiveness of smooth muscle of the canine basilar artery to EDRF and nitric oxide. AB - Experiments were designed to compare the reactivity of canine basilar arteries to endothelium-derived relaxing factor (EDRF) and nitric oxide. Preparations with endothelium activated by bradykinin were used to study relaxations induced with EDRF, whereas the inhibitory effect of nitric oxide was studied in preparations without endothelium. All experiments were performed in the presence of indomethacin. EDRF- and nitric oxide-induced relaxations were significantly augmented in the presence of superoxide dismutase plus catalase but were reduced in the presence of methylene blue, LY 83583, and hemoglobin. M & B 22984 did not affect relaxations to either EDRF or nitric oxide. These results indicate that in the canine basilar artery EDRF is not an oxygen-derived free radical. The similar responsiveness of the basilar artery to EDRF and nitric oxide is consistent with the proposal that in the canine basilar artery nitric oxide is the factor. PMID- 2552841 TI - Attenuated coronary relaxation after reperfusion: effects of superoxide dismutase and TxA2 inhibitor U 63557A. AB - Previous studies demonstrate endothelium-dependent leukotriene D4 (LTD4)-induced relaxation of canine coronary arterial rings in vitro. We now show that coronary occlusion followed by reperfusion attenuates (P less than 0.01) the relaxation of canine coronary artery rings in response to LTD4 as well as acetylcholine (ACh), suggesting loss of endothelium-dependent coronary reactivity (n = 6 dogs). Since superoxide anions have been shown to cause breakdown of endothelium-derived relaxing factor (EDRF), we wondered whether treatment of dogs with superoxide anion scavenger superoxide dismutase (SOD) would modulate the effects of LTD4 and ACh on reperfused coronary artery rings. Indeed, treatment of dogs (n = 5) with SOD before coronary reperfusion resulted in preservation of LTD4- and ACh-induced relaxation of coronary rings. Treatment of another five dogs with selective thromboxane-synthetase blocker U 63557A before coronary reperfusion also resulted in preservation of coronary ring relaxation in response to LTD4 and ACh. To determine the mechanism of U 63557A-induced preservation of coronary reactivity, canine neutrophil superoxide anion generation in the presence of U 63557A was measured. Although U 63557A had no effect on superoxide anion generation in neutrophils alone, it markedly (P less than 0.02) inhibited superoxide anion generation in neutrophils in the presence of platelets, most likely via shunting of accumulated cyclic endoperoxide in platelets toward formation of prostacyclin, which inhibits neutrophil superoxide anion production. Thus SOD and U 63557A protect against loss of endothelium-mediated vascular relaxation by LTD4 and ACh after coronary occlusion and reperfusion. PMID- 2552842 TI - Dithiothreitol restores contractile function to oxidant-injured cardiac muscle. AB - Reperfusion injury in ischemic myocardium is caused partially by polymorphonuclear leukocyte oxygen free radicals, the most toxic of which may be hypochlorous acid (HOCl). This study shows that dithiothreitol (DTT), a disulfide reducing agent, can restore contractile function to cardiac muscles that had been exposed to physiological levels of HOCl. Isometrically contracting isolated rat papillary muscles which were exposed to HOCl (300 microM) showed a rapid and essentially complete loss of developed force, an increase in resting force, and a sharp decline in myocyte protein sulfhydryls (PSH). The addition of DTT (1 mM) after 40 min resulted in a significant (40%) restoration of contractile function. Earlier addition of DTT effected a more complete functional recovery. The DTT induced recovery was accompanied by a matching increase in cellular PSH levels, suggesting that HOCl injury may be caused primarily by the oxidation of cysteine residues. These data suggest that DTT may prove to be useful in reversing oxidant injury in tissues exposed to oxygen free radicals. PMID- 2552843 TI - ACTH-(4-10) through gamma-MSH: evidence for a new class of central autonomic nervous system-regulating peptides. AB - Examination of the cardiovascular effects produced by peripheral administration of peptide sequences derived from adrenocorticotropic hormone (ACTH) led to the discovery of the pressor, cardioaccelerator, and natriuretic actions of intravenous (iv) ACTH-(4-10). Based on pharmacological studies in rats with alpha and beta-adrenergic receptor antagonists, the cardiovascular effects of this peptide appeared to be mediated by the release of catecholamines. A peptide sequence analogous to ACTH-(4-10), gamma-melanocyte-stimulating hormone (gamma MSH), possesses greater than 100-fold more cardiovascular activity and 1,000-fold more natriuretic activity than ACTH-(4-10). The pressor effect of iv gamma-MSH peptides appears to be dependent on the maintenance of preganglionic sympathetic drive, with no significant contribution of circulating vasopressin or angiotensin II. However, the presence of central vasopressinergic, and perhaps angiotensinergic, pathways appears to be crucial for expression of the full pressor effect of circulating gamma-MSH. Further evidence for the potential importance of the central nervous system (CNS) in these cardiovascular effects was obtained from central lesion experiments and a comparison of intracarotid vs. intrajugular infusions. Structure-activity studies suggested that the cardiovascular effects of ACTH-(4-10) or gamma-MSH are dependent on an Arg hydrophobic amino acid sequence, located at or near their COOH-terminal. A similar requirement for biological activity is found in molluscan cardioexcitatory peptides, and the molluscan peptides have cardiovascular effects in rats, which resemble ACTH-(4-10) or gamma-MSH. This suggests that peptides of the gamma-MSH family are the pharmacological analogues, and perhaps the physiological homologues, of a cardioexcitatory family of peptides found in molluscs and birds. Elevated circulating levels of peptides derived from the NH2 terminal of pro-opiomelanocortin (POMC) have been found in psychological stress, cardiovascular distress, and hemorrhage. Increases in central sympathetic drive are common to all of these states. gamma-MSH peptides have been localized to POMC neurons in the arcuate nucleus and nucleus commissuralis of the rat. Projections from the latter nucleus innervate hindbrain vasomotor centers. Intraventricular administration of gamma-MSH produces prolonged elevation of mean arterial pressure. gamma-MSH peptides may provide a link between humoral and neurogenic mechanisms in cardiovascular regulation and could potentially be important neurotransmitters for central control of the cardiovascular system. PMID- 2552844 TI - An explanation for decreased ketogenesis in the liver of the obese Zucker rat. AB - These studies were undertaken to further characterize and explain the differences in hepatic fatty acid metabolism between lean and obese Zucker rats. It was shown that the rate of palmitate or octanoate oxidation and the inhibition of palmitate oxidation by malonyl CoA in mitochondria isolated from lean and obese Zucker rats were similar. Cytochrome oxidase activity was similar in lean and obese rat livers. It was found that the addition of cytosol from the obese rat liver inhibited palmitate oxidation by 20-30% in mitochondria isolated from lean or obese rat livers and thus reproduced the conditions observed in the intact cell. Increased concentrations of metabolites such as malonyl CoA and glycerophosphate in the liver of the obese rat are likely contributors to this inhibitory effect. These results are extrapolated to the intact cell and suggest that decreased hepatic fatty acid oxidation in the obese rat can be accounted for by cytosolic influences on the mitochondria. The decreased rate of fatty acid oxidation observed in the intact hepatocyte or perfused liver cannot be explained by a defect in the capacity of mitochondria to oxidize substrate or by a decrease in mitochondrial number in the obese rat liver. PMID- 2552845 TI - Plasma level, urinary excretion, and metabolic production of cGMP during gestation in rats. AB - We postulated that guanosine 3',5'-cyclic monophosphate (cGMP), a cellular mediator of vascular smooth muscle relaxation, might mediate maternal renal and cardiovascular hemodynamic adaptation to pregnancy. Because extracellular levels of cGMP most likely reflect intracellular production, we began our investigation of this hypothesis by measuring the plasma concentration, urinary excretion, and metabolic clearance rates of cGMP during pregnancy in rats. Plasma cGMP was significantly elevated during mid- and late pregnancy, whereas urinary excretion of cGMP was increased throughout pregnancy. The fractional excretion of cGMP by the kidneys was 0.90 +/- 0.15 in the nonpregnant condition. In contrast, plasma levels of adenosine 3',5'-cyclic monophosphate were unchanged during pregnancy, and its urinary excretion rose slightly, reaching significance only on gestational day 20. There was also a significant rise in urinary excretion of cGMP throughout pseudopregnancy. The metabolic clearance rate of cGMP measured in chronically instrumented rats before, during, and after pregnancy was not significantly altered during gestation. The elevated plasma level of cGMP during gestation in rats, in the face of an unchanged metabolic clearance, reflects augmented tissue(s) production of cGMP, although enhanced cellular efflux may contribute. Because cGMP is a second messenger for several vasodilatory hormones, our data are consistent with the hypothesis that vascular production of cGMP may increase during pregnancy and thereby contribute to maternal renal and cardiovascular vasodilation. (Most investigators have not observed increment of plasma atrial natriuretic peptide in rat gestation; therefore this hormone is an unlikely first messenger for the elevated extracellular levels of cGMP that we have observed. Finally, pseudopregnant rats also showed enhanced urinary excretion of cGMP, which suggests that the proliferative activity that accompanies fetoplacental maturation, as well as hormones elaborated by the fetoplacental unit, is not necessary for the rise in urinary excretion of cGMP observed during pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552846 TI - mu-Opioid peptide modulation of cardiovascular and sympathoadrenal responses to stress. AB - The effects of mu- and delta-opioid receptor activation on sympathoadrenal and cardiovascular responses to stress were examined in conscious rats. The mu selective agonist [D-Ala2,N-Me-Phe4,Gly5-ol]enkephalin (DAGO) or the delta selective agonist [D-Pen2,D-Pen5]enkephalin (DPDPE) was injected into a lateral cerebral ventricle, then rats were stressed by restraint. Plasma catecholamines were measured, and arterial blood pressure and heart rate were recorded continuously. Restraint stress evoked increases in plasma catecholamines and heart rate in saline-pretreated rats. Both DAGO and DPDPE increased basal plasma levels of catecholamines and blood pressure, and DAGO, 5 nmol, produced bradycardia. DAGO, 5 nmol, but not DPDPE, potentiated the plasma catecholamine responses to restraint. However, the presence of DAGO or DPDPE during restraint resulted in decreases in heart rate and blood pressure. The effects of DAGO and DPDPE on plasma catecholamines, heart rate, and blood pressure were blocked by a mu-selective dose of naloxone but were not reversed by the delta-selective antagonist ICI 174864. These results indicate that mu-receptor stimulation during restraint stress facilitates sympathoadrenal and parasympathetic outflow and results in vasodilatation of some peripheral vascular beds. PMID- 2552847 TI - Electrogenic sodium-proton exchange in membrane vesicles from crab (Carcinus maenas) gill. AB - Plasma membrane vesicles from Na+-absorbing posterior gills of the green crab Carcinus maenas were shown to contain a Na+-H+ antiporter sensitive to amiloride (Ki = 280 mumol/l), using a fluorometric assay based on acridine orange. Lithium was found to support Li+-H+ exchange, giving Michaelis-Menten kinetics (K0.5 = 20 mmol/l) and a Hill coefficient of 1.32 +/- 0.06, indicating a stoichiometry of 1 Li+:1 H+. The stoichiometry of Na+-H+ exchange, however, appeared to be 2 Na+:1 H+. The dependence of exchange rate on Na+ concentration followed a sigmoid relationship (K0.5 = 34 mmol/l), producing a Hill coefficient of 1.99 +/- 0.18. Measurements made with the potential-sensitive dye 3,3' dipropylthiadicarboxocyanine iodide directly demonstrated electrogenicity of Na+ H+ exchange. Na+-loaded vesicles showed greater potential change upon addition to Na+-free medium than upon addition to Na+-containing medium, and the developed potential was reversed by the addition of external Na+. Amiloride reduced the rate of membrane polarization and abolished the reversal by external Na+. The crustacean Na+-H+ antiporter therefore bears similarities to the vertebrate antiporter but is uniquely electrogenic. PMID- 2552848 TI - The pathology of Cronkhite-Canada polyps. A comparison to juvenile polyposis. AB - We studied Cronkhite-Canada (CC) polyps from nine patients, and compared them to gastric and colonic juvenile and gastric hyperplastic polyps. The CC polyp is characterized by its broad sessile base, expanded edematous lamina propria, and cystic glands. Similar features are found in the lesions of juvenile polyposis and gastric hyperplastic polyps. The only reliable distinction between CC and colonic juvenile polyposis was the pedunculated growth of the latter; however, this feature did not hold for gastric lesions. Unlike CC polyps, juvenile polyps sometimes have areas of dysplasia, but this is not typical. Therefore the diagnosis of CC polps, especially when located in the stomach, requires the presence of the ectodermal changes characteristic of this syndrome. PMID- 2552849 TI - Ultrastructure of experimental intestinal invasive amebiasis. AB - The morphological features of early intestinal ulcerations induced in rodents with axenic cultures of Entamoeba histolytica were studied by scanning and transmission electron microscopy. Amebas did not attach to the luminal surface of the mucosa except at interglandular regions, where parasites penetrated apparently through pseudopodial movement. Once in the lamina propria, trophozoites multiplied and destroyed mucosal components. Damage extended laterally through the mucosa, but progression to deeper layers of the intestinal wall was prevented by the muscularis mucosae, which acted as a partial barrier. This was eventually breached at focal points where amebas invaded the submucosa. Electron microscopy clearly showed the lysis of abundant polymorphonuclear neutrophil leukocytes (PMNs) by the amebas at the periphery of intestinal ulcerations as well as the lack of bacteria at these sites. This study demonstrates that recruitment and destruction of inflammatory cells following intestinal amebic invasion may take place in the absence of bacterial multiplication. The observations provide histological support to the hypothesis that lysis of PMNs by trophozoites participates in the genesis of amebic intestinal lesions. PMID- 2552850 TI - Visceral leishmaniasis unresponsive to pentostam caused by Leishmania tropica in Kenya. AB - We report the characterization of 6 Leishmania tropica isolates from 2 patients with visceral leishmaniasis who were unresponsive to treatment with sodium stibogluconate. The Leishmania isolates, MHOM/KE/81/NLB-029A, -029XIB, and 029XIC and MHOM/KE/81/NLB-030I, -030B, and -030XXA, all from splenic aspirates, were characterized by cellulose acetate electrophoresis using 11 enzymes: malate dehydrogenase, malic enzyme, phosphogluconate dehydrogenase, glucose-6-phosphate dehydrogenase, superoxide dismutase, glutamate-oxaloacetate transaminase, adenylate kinase, nucleoside hydrolase, mannose phosphate isomerase, glucose phosphate isomerase, and phosphoglucomutase. Isozyme migration patterns were indistinguishable from those of 2 WHO reference strains of Leishmania tropica (MHOM/SU/60/LRC-L39, NLB-305 and MHOM/IQ/OO/LRC-L36, NLB-067). These are the first reported cases of visceral leishmaniasis (kala-azar) caused by L. tropica in Africa; these cases were refractory to sodium stibogluconate. PMID- 2552851 TI - An in vitro micromethod for drug sensitivity testing of Leishmania. AB - We describe an in vitro microtest which is quantitative, rapid, and readily applicable to parasites isolated from all major forms of human leishmaniasis. It is based on drug-mediated inhibition of promastigote catabolism of a battery of simple 14C-substrates to 14CO2. Each test requires less than 1 microCi. The test is conducted in a serum-free, chemically defined medium containing 120 micrograms protein/ml, minimizing the possible interference of drug: serum protein interaction. Prior adaptation is not necessary to cultivate "difficult-to-grow" species. Leishmania sensitivity to pentavalent antimonials is detectable at micrograms levels below concentrations achievable in patients' sera. PMID- 2552852 TI - Amebiasis now. PMID- 2552853 TI - Oncologic and functional considerations of total glossectomy. AB - The efficacy of total glossectomy for advanced carcinoma of the tongue remains controversial. A retrospective chart review was undertaken to evaluate the oncologic and functional results in 17 consecutive patients who underwent this procedure. There were two patients with stage III disease, eight with stage IV disease, and seven with recurrent disease. The larynx was preserved in seven patients. One patient required a secondary laryngectomy. All patients were reconstructed immediately, 11 with a pectoralis major myocutaneous flap and 6 with free-tissue transfer. The operative mortality was 6 percent; the morbidity was 59 percent. At last follow-up, 53 percent of the patients were alive without disease, with a mean disease-free survival period of 36 months. Ninety-three percent of the patients regained swallowing and independent oral alimentation; 80 percent of those with laryngeal preservation regained intelligible speech. We have concluded that total glossectomy should be considered as a primary modality for advanced carcinoma of the tongue and not solely reserved for salvage in hopeless situations. With or without laryngectomy, excellent survival and functional results can be obtained. PMID- 2552854 TI - Microvascular approach to function and appearance of large orbital maxillary defects. AB - Forty-three patients with major three-dimensional orbitomaxillary defects underwent 48 free-tissue transfers for defects arising from resection of a neoplasm or trauma. Defects were complex, involving the malar skin, mandible, lateral nasal wall, orbit, palate, and brain. A three-dimensional approach attempting to recreate the midface by folding the flaps was the usual method, with subsequent revisions if necessary. Ten patients had simultaneous free bone grafts and 6 had vascularized bone grafts. There were three flap failures and four perioperative deaths. Excluding the 4 deaths, the results were assessed as excellent in 25 patients, good in 12, and poor in 2. Free-tissue transfer is a reliable method of providing enough tissue at one initial operation to satisfy complex three-dimensional defects of the orbitomaxillary region. Followed by minor revision, it provides a means of restoring function and appearance without multiple staged procedures with a high risk of failure. PMID- 2552855 TI - An overview of clinical trials of low molecular weight heparin fractions. PMID- 2552856 TI - [The role of surgery and low molecular weight heparin in the treatment of cerebral venous thrombosis. Apropos of 2 cases]. AB - For one case of thrombosis of sinus sagittalis superior joint haemorrhagic cerebral softening necessitates surgical draining of haemorrhagic focus grown to a true intracranial haematoma; for the another case direct low molecular weight heparin treatment obtained a mere evolution. Authors are of the opinion that early low molecular weight heparin treatment is able to avoid massive haemorrhage. PMID- 2552857 TI - [Clinical tolerance of CY 216 (Fraxiparin) in the prevention of thromboembolic accidents after neurosurgery]. AB - Clinical tolerance of CY 216 was assessed on 97 patients in two neurosurgical departments (Hopital Lariboisiere and Hopital Beaujon, Paris). All the 97 patients were operated on by different cranial approaches, but always including bone flap. No thrombo-embolic event could be detected in this series. Post operative intracranial hemorrhage was observed in 19 cases on the basis of CT scan performed on the first and seventh post-operative day. Among these 19 patients, 8 presented an increase of the hemorrhagic aspect between the 2 CT scan. However, clinical state of these patients improved and no reoperation was necessary to treat these hemorrhages. From this study, we consider that CY 216 is an effective mean of thrombo-embolism prevention with little hemorrhagic side effect. Therefore, it can be recommended in the aftermath of intracranial surgical procedures. PMID- 2552858 TI - [Current methods of birth control]. PMID- 2552859 TI - [Dynamic gamma-camera scintigraphy as a method for evaluating the function of the ovarian parenchyma]. AB - A dynamic gamma-camera scintigraphy is proposed as a combined method, providing morphological and functional picture for the presence of eventual pathological changes in the region of ovaries. After detailed description of the method results from the examination of 45 women are presented: 35 suffered from diseases, involving adnexal region and 10--are controls. It is established that normal ovaries are visualized medially from the large iliac vessels as clear, ovally formed homogeneous cumulation of radionuclide. No such cumulation is observed after lack of ovarian parenchyma or after not well made perfusion. The method has serious advantages, when anomalies in the development of ovaries are search for as well as in diseases, changing form and size of the ovary (deformation, lack of homogeneity, ect.). A method is proposed for its advantage that it is not invasive, the examination is performed easily and is safe for patients. PMID- 2552860 TI - [Dynamic gamma-camera excretory scintigraphy in the diagnosis of a rare anomaly in the development of the internal genitalia (the Slotnik-Goldfarb syndrome with uterus unicornis)]. AB - The combination of a missing ovary and uterine tube on the one side (the syndrome of Slotnik-Goldfarb) with ipsilateral absence of the uterine horn is a rare anomaly, undescribed in our obstetric-gynecological literature. The difficulties in the diagnosis of this anomaly are connected with the necessity to use invasive methods for establishment of respective changes. The method was used in a woman, suffering from sterility, in connection with examining the possibilities of dynamic gamma-camera scintigraphy to discover ovarian parenchyma. Hysterosalpingography proved the presence of a hypoplastic uterine horn on the right side, an uterine tube on the same side (changed and nonpatent). Dysovulatory syndrome was established in this woman clinically. The performed scintigraphy confirmed the presence of ovarian parenchyma on the right side as well as a hypoplastic uterine horn and uterine tube and complete lack of accumulation of radionuclide on the left side, which suggested absence of ovarian parenchyma in the sidee in which the uterine horn and uterine tube were undeveloped as well. It is evaluated that the scintigraphic technique, described by the authors, could be used for establishment of such rare anomalies by noninvasive way. PMID- 2552861 TI - Human neutrophils are not severely injured in conditions mimicking social drinking. AB - In an attempt to assess the effect of alcohol per se on human polymorphonuclear neutrophils (PMN), irrespective of other physiopathological parameters, we examined neutrophil function in healthy volunteers who had taken a single large dose of whisky. Before and at different times after ingestion, several PMN properties were simultaneously tested including random migration, in vitro chemotaxis, adherence, aggregation, cytochrome C reduction, phagocytosis, bacterial killing, intracellular cAMP and cGMP contents, myeloperoxidase, and neutrophil alkaline phosphatase scores. Only phagocytosis of Staphylococcus aureus was significantly depressed after alcohol ingestion. Adherence was inhibited only in some individuals when their respective blood alcohol levels were the highest. Both alterations were moderate and reversible. These data point out the limited effect of occasional alcohol consumption on the different facets of neutrophil behavior. These findings suggest that factors other than alcohol itself could be concerned in the marked PMN dysfunction well established in chronic alcoholism. PMID- 2552862 TI - Pathological changes in peripheral nerves in experimental fetal alcohol syndrome. AB - The peripheral nerve pathology is described in experimental fetal alcohol syndrome. The marked retraction and shrinkage of axons with vacuolar spaces in the periaxonal area are the striking features. The direct toxicity of ethanol or its metabolites on the nerve fibers is considered to be the cause for pathological changes. Additional biochemical factors, pyridoxal phosphate dependent enzyme-depletion in the genesis of nerve damage, is proposed. PMID- 2552863 TI - Prenatal exposure to ethanol alters the ontogeny of the beta-endorphin response to stress. AB - To determine whether prenatal exposure to ethanol alters the response of the beta endorphin (beta-EP) system to stress, the effect of two types of stressful stimuli, ether and cold, was examined in the offspring of rats which during pregnancy were: (a) fed with an ethanol-containing diet; (b) pair-fed with an isocaloric sucrose diet; and (c) fed ad libitum with standard lab chow (basic control group). The effect of stress on the content of beta-EP in the serum, pituitary gland and hypothalamus, as well as on the serum corticosterone and hypothalamic corticotropin-releasing factor (CRF) content was examined. Pups prenatally exposed to ethanol had significantly higher serum beta-EP levels on Day 1 and higher serum corticosterone levels on Days 1-3 when compared to their pair-fed or basic controls. On all days tested pituitary beta-EP content was lower in the offspring of the ethanol-treated rats than in the control groups. There was no difference in the total hypothalamic beta-EP content between the three treatment groups; however, during the first 10 days of life a higher concentration (ng/mg protein) of beta-EP was observed in the hypothalami of the ethanol and the pair-fed group when compared to the basic control pups. Hypothalamic CRF levels, though significantly lower in the pups exposed to ethanol in utero than in the control groups on Day 3, increased significantly in the ethanol group between Days 14 and 22, while no significant change was observed during this period in either of the control groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552864 TI - Immunohistochemical study on the development of the adenohypophysial cells in the lizard Gallotia galloti. AB - Immunohistochemical methods have been used to study the embryonic and postnatal development of the hormone-producing cells in the adenohypophysis of the lizard Gallotia galloti. In this species, Rathke's pouch is formed between stages 30 to 32 of the embryonic development, although the first sign of immunoreactivity to antisera against adenohypophysial hormones occurs in stage 33 in the pars distalis anlage. These cells derive from the dorsal face of Rathke's pouch and are immunoreactive to anti-ACTH serum. The cytodifferentiation of ACTH and MSH cells occurs in the pars intermedia in stage 34. The TSH cells appear at stage 35 and the gonadotrope cells at stage 37. These cells derive from both the dorsal and ventral face of the Rathke's pouch. The LTH cells are revealed at stage 39 and are only originated from the dorsal face. The STH cells, which come from the dorsal as well as ventral face, are the last secretory cells differentiated just before hatching. During postnatal development an increase and also a redistribution of the immunoreactive cells occur until acquiring the adult distribution. PMID- 2552865 TI - Changes in epithelial secretory cells and potentiation of neurogenic inflammation in the trachea of rats with respiratory tract infections. AB - In rats respiratory tract infections due to Sendai virus and coronavirus usually are transient, but they can have long-lasting consequences when accompanied by Mycoplasma pulmonis infections. Morphological alterations in the tracheal epithelium and a potentiation of the inflammatory response evoked by sensory nerve stimulation ("neurogenic inflammation") are evident nine weeks after the infections begin, but the extent to which these changes are present at earlier times is not known. In the present study we characterized these abnormalities in the epithelium and determined the extent to which they are present 3 and 6 weeks after the infections begin. We also determined the magnitude of the potentiation of neurogenic inflammation at these times, whether the potentiation can be reversed by glucocorticoids, and whether a proliferation of blood vessels contributes to the abnormally large amount of plasma extravasation associated with this potentiation. To this end, we studied Long-Evans rats that acquired these viral and mycoplasmal infections from other rats. We found that the tracheal epithelium of the infected rats had ten times as many Alcian blue-PAS positive mucous cells as did that of pathogen-free rats; but it contained none of the serous cells typical of pathogen-free rats, so the total number of secretory cells was not increased. In addition, the epithelium of the infected rats had three times the number of ciliated cells and had only a third of the number of globule leukocytes. In response to an injection of capsaicin (150 micrograms/kg i.v.), the tracheas of the infected rats developed an abnormally large amount of extravasation of two tracers, Evans blue dye and Monastral blue pigment, and had an abnormally large number of Monastral blue-labeled venules, particularly in regions of mucosa overlying the cartilaginous rings. This abnormally large amount of extravasation was blocked by dexamethasone (1 mg/day i.p. for 5 days). We conclude that M. pulmonis infections, exacerbated at the outset by viral infections, result within three weeks in the transformation of epithelial serous cells into mucous cells, the proliferation of ciliated cells, and the depletion of globule leukocytes. They also cause a proliferation of mediator-sensitive blood vessels in the airway mucosa, which is likely to contribute to the potentiation of neurogenic inflammation that accompanies these infections. PMID- 2552866 TI - Role of spinal adenosine receptors in modulating the hyperesthesia produced by spinal glycine receptor antagonism. AB - The intrathecal administration of strychnine in rats yields a prominent touch evoked allodynia. The effects of an intrathecally administered A1 adenosine agonist: N6-(L-2-phenylisopropyl)-adenosine (LPIA) or an A2 adenosine agonist: 5' (N-ethyl carboxamido)-adenosine (NECA), on this touch-evoked hyperesthesia were examined. Over the range of 0.3-1.0 nmol these agents produced a dose-dependent inhibition of the strychnine-evoked hyperesthesia. This inhibition was reversed following intraperitoneal injection of caffeine, an adenosine receptor antagonist. No statistical differences between LPIA and NECA were recorded. The powerful effect of adenosine analogues on strychnine hyperesthesia occur at doses that have only a mild analgesic effect on the thermally evoked hot-plate response. This effect is in contrast to opioids, which have been reported to be only minimally effective against strychnine-evoked hyperesthesia. The characteristics of the strychnine hyperesthesia appear to mimic the clinical phenomenon observed in patients suffering from sensory dysesthesia following nerve injury and suggest a possible role for the adenosine receptor in certain pain states. PMID- 2552867 TI - Postoperative isolated dysfunction of the long thoracic nerve: a rare entity of uncertain etiology. AB - A "winged" scapula is a rare, poorly understood, and potentially disabling curiosity following anesthesia and surgery. It is produced by dysfunction of the long thoracic nerve and consequent paralysis of the serratus anterior muscle. A survey of senior anesthesiologists indicated a consistent lack of familiarity with the entity. This article presents six cases of postoperative long thoracic nerve palsy. In a literature review of 111 instances of long thoracic nerve palsy, 51 were trauma-related, 47 were either idiopathic or of debatable origin, and 13 appeared following a surgical or obstetrical procedure. Unprovable etiologic contentions were frequent. Considerations of the etiologies of postoperative long thoracic nerve palsies must include a coincidental infectious neuropathy ("neuralgic amyotrophy") as a valid alternative to the assertion that a preventable injury occurred during anesthesia. PMID- 2552868 TI - Dexamethasone isonicotinate inhibits dual and late asthmatic reactions but not the increase of airway responsiveness induced by toluene diisocyanate in sensitized subjects. AB - To determine whether treatment with aerosolized dexamethasone isonicotinate inhibits asthmatic reactions and the associated increase in airway responsiveness induced by toluene diisocyanate (TDI), we studied six sensitized subjects with previously demonstrated dual or late asthmatic reaction after inhalation challenge with TDI. Dexamethasone isonicotinate (four puffs bid for seven days, ie, 0.5 mg bid for seven days; last four puffs 30 minutes before TDI) was administered for seven days before the inhalation challenge with TDI (0.010 to 0.015 ppm for 10 to 30 minutes) to each subject, according to a single-blind study design. When the subjects received no treatment, FEV1 markedly decreased and airway responsiveness increased after exposure to TDI. By contrast, when the subjects were treated with dexamethasone-isonicotinate, FEV1 decreased significantly less, but airway responsiveness still significantly increased after exposure to TDI. These results suggest that aerosolized dexamethasone isonicotinate may be used in the prophylaxis of TDI-induced late asthmatic reactions. PMID- 2552869 TI - Encephalopathy as a presentation of pediatric AIDS: case report. AB - Fifty to ninety percent of pediatric AIDS cases are complicated by neurologic dysfunction. We present a case of a 5-year-old black female with AIDS encephalopathy and Mycobacterium avium intracellulare. Her initial presentation was that of neuroencephalopathy with loss of developmental milestones, pyramidal tract signs, and subsequent evidence of cortical atrophy. Her initial CT scan at the time of frank encephalopathy was normal, whereas 18 months into the clinical course of her encephalopathy, her CT scan of the head demonstrated typical ventricular dilatation and severe cortical atrophy consonant with her clinical picture. She subsequently developed Mycobacterium avium intracellulare documented by gastric aspirate culture and other opportunistic infections including Candida esophagitis. Her neuroencephalopathy plateaued with continued evidence of immune dysfunction and mycobacterium by gastric aspirate, despite triple antibiotic therapy with INH, streptomycin, Pyrazinamide with later addition of Rifampin and final substitution of the investigational congener Rifabutin. AIDS encephalopathy and Mycobacterium intracellulare are discussed in terms of their prognosis and therapy, particularly in view of new reports of the application of AZT and immunoglobulin therapy. PMID- 2552870 TI - Severe metabolic acidosis after acute naproxen sodium ingestion. AB - Naproxen, a nonsteroidal anti-inflammatory drug (NSAID), is a propionic acid derivative that possesses analgesic and antipyretic properties through inhibition of prostaglandin synthesis. The propionic acids have been considered the least toxic of the NSAIDs, and one, ibuprofen, is currently available as an over-the counter medication. Though acidosis has been reported with ibuprofen, no such occurrence has been reported for naproxen sodium. We report the case of a 15-year old girl who presented with severe metabolic acidosis and seizures that rapidly followed naproxen sodium ingestion. Serum bicarbonate levels returned to normal 12 hours after admission and correlated with the known pharmacokinetics of naproxen. The pharmacokinetics of naproxen and treatment of its overdose, and possible mechanisms of metabolic acidosis are reviewed. PMID- 2552871 TI - Latent infection and subsequent reactivation of pseudorabies virus in swine exposed to pseudorabies virus while nursing immune dams. AB - The ability of pseudorabies virus (PRV) to infect and establish latency in pigs with passively acquired (maternal) antibody for PRV was tested by exposing such pigs to the virus and subsequently attempting to reactivate latent virus by administering large doses of dexamethasone. Pigs of each of 4 litters that had nursed gilts with relatively high (512, gilts 1 and 2), moderate (32, gilt 3), and no (less than 2, gilt 4) serum titers of virus-neutralizing (VN) antibodies for PRV were allotted to 3 treatment groups (A, B, C) when they were 2 weeks old. Group-A pigs were separated from littermates and dam and thereafter kept in isolation; group-B pigs were experimentally exposed oronasally to PRV and 1 hour later returned to their dam; group-C pigs were kept with their dam and potentially exposed to PRV by contact with littermates of group B. Sera obtained from pigs at selected intervals until they were 17 weeks old were tested for VN activity and for precipitating activity for radiolabeled viral proteins. All group-A pigs remained clinically normal throughout the experiment. Depending on the initial amount of passively acquired antibody, little or no serum VN or precipitating activity remained by the time these pigs were 17 weeks old. Group-B and -C pigs, with relatively high amounts of passively acquired antibody when exposed to PRV, also remained clinically normal. However, most became latently infected as subsequently evidenced by either dexamethasone-induced or noninduced virus reactivation. Noninduced reactivation may have been initiated by weaning the pigs when they were about 8 weeks old.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552872 TI - Synergistic antiviral activities of acyclovir and recombinant human leukocyte (alpha) interferon on feline herpesvirus replication. AB - The antiviral activities of 9-(2-hydroxyethoxymethyl)guanine (acyclovir; ACV) either alone or combined with recombinant human leukocyte (alpha) A/D interferon (rHuIFN-alpha) against feline herpesvirus type 1 (FHV-1) were evaluated in feline embryo cell cultures, using an infectivity-inhibition assay. In ACV-treated cultures, the 50% inhibitory dose (ID50) was approximately 10 to 20 micrograms of ACV/ml. Maximal inhibition of FHV-1 infectivity (range, 3.4 to 4.2 log10 TCID50) was observed when high test doses of ACV (125 or 250 micrograms/ml) were given 1 to 6 hours after infection. Although mild inhibition (range, 0.3 to 1.6 log10 TCID50) of virus was observed at lower drug doses (10 to 62.5 micrograms/ml), FHV 1 was relatively resistant to ACV and required higher minimal inhibitory doses than those reported for other herpesviruses. However, when ACV was combined with 10 or 100 U of rHuIFN-alpha/ml, synergistic antiviral effects were associated with ACV dosage of 10 to 62.5 micrograms/ml. Antiviral activities resulting from use of the combined drugs permitted nearly eightfold reduction in the dose of ACV required to achieve maximal inhibition of FHV-1. Significant (P less than 0.01) synergistic interactions with ACV resulted when the rHuIFN-alpha was given before or after infection; at the lower doses of ACV, however, rHuIFN-alpha pretreatment was more effective. Although dosages of either greater than or equal to 62.5 micrograms of ACV/ml or 100 U of rHuIFN-alpha/ml were cytosuppressive in control cell cultures, additive anticellular effects were not observed at synergistic combinations of ACV and 10 U of rHuIFN-alpha/ml. PMID- 2552873 TI - Molecular cloning of serogroup- and serotype-specific genome segments from bluetongue virus serotype 11. AB - Genome segments 2, 6, 8, and 9 of bluetongue virus (BTV) serotype 11, coding for P2, NS1, NS2, and P6, respectively, were cloned into pUC 8. Sizes of segment-2 and segment-6 clones indicated partial copies (55% and 80% of full length, respectively), whereas segment 8 and 9 clones represented full-length copies. Northern blot hybridizations of the clones to the 5 United States BTV prototypic serotypes (2, 10, 11, 13, and 17) revealed segment-2 clone to be serotype specific to BTV-11, whereas segment 6, 8, and 9 clones were able to detect all serotypes to varying degrees. All clones failed to detect the related orbivirus, epizootic hemorrhagic disease virus. PMID- 2552874 TI - Transmission of bovine leukemia virus by Tabanus fuscicostatus. AB - Bovine leukemia virus (BLV) was transmitted by horse flies, Tabanus fuscicostatus, from a cow with a lymphocyte count of 31,500/mm3 to goats and dairy calves. As few as 10 and 20 flies transmitted BLV to goats and calves respectively, but the minimal number of flies required to transmit the infection was not established. Groups of 150 and 100 T fuscicostatus transmitted BLV to beef calves from a cow with a lymphocyte count of 14,600/mm3. These results support a role for horse flies in the horizontal transmission of BLV. PMID- 2552875 TI - Cross-protective immunity between equine encephalomyelitis viruses in equids. AB - Eighteen equids were inoculated with eastern equine encephalomyelitis (EEE) and 18 equids with western equine encephalomyelitis (WEE) viruses to produce EEE virus- and WEE virus-immunized equids. Twelve surviving EEE virus-seropositive equids, 15 surviving WEE virus-seropositive equids, and 10 nonimmunized, seronegative equids (controls) were subsequently inoculated with an equine pathogenic (epizootic) strain of Venezuelan equine encephalomyelitis (VEE) virus to determine cross-protective immunity. Challenge infection produced 90% mortality in control (nonimmunized) equids, and 40% mortality in WEE virus seropositive equids; all EEE virus-seropositive equids survived. Postchallenge exposure VEE viremia levels in EEE virus- or WEE virus-seropositive equids were lower than those in the 10 nonimmunized VEE virus-inoculated control equids. Plaque-neutralizing antibody responses to VEE virus in the EEE virus- and WEE virus-seropositive equids were similar in time of onset and titer to the antibody responses of nonimmunized equids. Neutralizing antibody to the third equine encephalomyelitis virus (either EEE virus or WEE virus) was detectable in 19 of 27 equids after inoculation with the challenge virus, VEE. Demonstration of cross protective immunity between EEE or WEE virus and VEE virus in equids confirmed field observations made during the VEE epizootic in Texas in 1971. PMID- 2552876 TI - Prevalence of antibodies to alcelaphine herpesvirus-1 and nucleic acid hybridization analysis of viruses isolated from captive exotic ruminants. AB - A serologic survey was conducted to determine the prevalence of antibodies to alcelaphine herpesvirus-1 (AHV-1) in captive exotic ruminants within the United States. Forty-six percent of the members of the subfamily Alcelaphinae (wildebeest, topi, hartebeest) in the family Bovidae had virus-neutralizing antibody to AHV-1. Other subfamilies of Bovidae with high prevalence of virus neutralizing antibodies to AHV-1 included Hippotraginae (oryx and addax) and Caprinae (sheep and goats), with prevalence of 45% and 29%, respectively. Herpesviruses that have been isolated from captive exotic ruminant species, including healthy animals and those with clinical malignant catarrhal fever at the Oklahoma City Zoo and the San Diego Zoo/Wild Animal Park, were analyzed by DNA restriction enzyme analysis and blot hybridization. Variation has been detected among the genomes of several malignant catarrhal fever virus isolates obtained from various exotic species of ruminants, using the DNA restriction enzymes BamHI and HindIII. The DNA of these virus isolates is distinct from that of bovine herpesviruses 1, 2, and 4, as demonstrated by restriction enzyme analysis and nucleic acid hybridization. On the basis of restriction enzyme analysis and nucleic acid hybridization data, the DNA from each of the putative alcelaphine herpesvirus isolates examined, except for the topi virus isolate, had a high degree of DNA sequence similarity with the original AHV-1 isolate, WC-11, from a blue wildebeest. PMID- 2552877 TI - Differentiation of avian adenovirus type-II strains by restriction endonuclease fingerprinting. AB - Three serologically indistinguishable viruses from the avian adenovirus type-II splenomegaly virus of chickens, marble spleen disease virus of pheasants, and hemorrhagic enteritis virus of turkeys, were analyzed by restriction endonuclease fingerprinting. The DNA from these viruses were examined with 6 restriction endonucleases (Bgl II, EcoRI, HindIII, Hha I, Xho I, and BamHI). Markedly different DNA cleavage patterns were found in these virus isolates with all the 5 enzymes, except with BamHI, suggesting genetic differences between isolates of adenovirus type II. Restriction endonuclease analyses were found to provide a method for distinguishing genetically different, and yet serologically similar, strains of avian adenovirus type II. PMID- 2552878 TI - Influence of temperature and age on the recovery of pseudorabies virus from houseflies (Musca domestica). AB - In laboratory experiments, pseudorabies virus was readily recovered from within the body of houseflies (Musca domestica L) that had ingested the virus. Age of the fly and ambient temperature affected the rate of virus inactivation within the houseflies. Virus half-life in 3-day-old flies was 6.36 hours vs 2.81 hours in flies 8 or 13 days old. Half-life in 5-day-old flies was 12.92 hours at 10 C, 5.95 hours at 20 C, and 2.69 hours at 30 C. Virus half-life in dead flies was 9.06 hours at 10 C, 4.28 hours at 20 C, and 1.71 hours at 30 C. The data did not provide any evidence of virus replication in either living or dead flies. PMID- 2552879 TI - Modulation of the cellular immune responses to T-cell-dependent and T cell independent antigens in lambs with induced bovine viral diarrhea virus infection. AB - Functional interaction between lymphoid cells and lymphotropic viruses is particularly evident for bovine viral diarrhea virus (BVDV) in cattle and its closely related virus, the border disease virus (BVDV) in sheep. The most important aspect of acute or chronic phases of BVDV or BDV infection was the host's increased susceptibility to secondary bacterial or viral infection. To study the ability of BVDV to alter the development of the cellular immune responses to concomitant inoculation with T cell-dependent and T cell-independent antigens, lambs were inoculated twice with rabbit RBC and Escherichia coli lipopolysacharide (LPS) and then were infected with a cytopathic strain of BVDV at postinoculation day 3. Leukopenia characterized by lymphopenia developed after BVDV infection. Increased [3H]thymidine incorporation was observed in resting or lectin-stimulated blood mononuclear cells in the first weeks after inoculation in BVDV-infected lambs, but was followed by decreased [3H]thymidine incorporation after the second inoculation for up to 8 weeks after initial inoculation. In contrast, transient decrease of blastogenic responses, associated with toxic effect of LPS, was detected in inoculated noninfected lambs, but was followed by stimulation of cellular immune responses. Inoculated noninfected lambs had good in vitro cellular immune response to rabbit RBC and LPS antigens, whereas lymphocytes from BVDV-infected lambs could not mount lasting cellular immune responses to antigens or BVDV. Results suggest that BVDV infection in lambs modulates the ability of lymphocytes to respond to lectins or antigenic stimuli according to the time after infection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552880 TI - Fiber digestion and voluntary intake in horses after adaptation to extensive large-colon resection. AB - Each of 3 digestion trials (3 forage diets) was performed on 2 groups of horses 6 to 12 months after sham operation (group 1; n = 3) or large-colon resection (group 2; n = 5). Diets were alfalfa pellets, alfalfa hay, and grass hay. Feed and fecal analyses were performed to determine apparent digestion of dry matter, organic matter, and crude protein and true digestion of dry matter, organic matter, crude protein, total plant cell wall, hemicellulose, cellulose, and lignin. Additional fecal and metabolic variables determined were percentage of fecal water, total fecal water, metabolic organic matter, metabolic crude protein, and metabolic nitrogen. Large-colon resection decreased the digestion of plant cell wall because of decreased digestion of cellulose in alfalfa pellet and grass hay diets, but not in alfalfa hay diet. Insufficient digestible energy and/or protein from grass hay was obtained by horses with colon resection, and significant (P less than 0.05) weight loss was observed. Voluntary intake was significantly (P less than 0.05) increased by horses with colon resection. Of the diets studied, alfalfa hay was the most appropriate forage diet, compared with average grass hay and alfalfa pellet diet, for horses after extensive large-colon resection. Additionally, horses with colon resection may have higher levels of feed intake than do horses without colon resection. PMID- 2552881 TI - Providing mental health care services to Americans. PMID- 2552882 TI - GTP gamma S increases thrombin-mediated inositol trisphosphate accumulation in permeabilized human endothelial cells. AB - Ca2+-mobilizing agonists stimulate phospholipase C-mediated phosphatidylinositol 4,5-bisphosphate hydrolysis and inositol trisphosphate (IP3) formation in pulmonary as well as in peripheral vascular endothelial cells (EC). In general, it is believed that receptor-phospholipase C interactions involve a guanine nucleotide regulatory (G) protein. This interaction can be inhibited by Bordetella pertussis toxin in certain cells. Here we report that pertussis toxin catalyzes the [32P]ADP ribosylation of a Mr = 41,000 protein in human umbilical vein EC. However, prior EC treatment with pertussis toxin (250 ng/ml for 20 h) does not inhibit thrombin-induced Ca2+ flux or IP3 formation, despite markedly attenuating the radiolabeling of the Mr = 41,000 protein (less than 5% control). Treatment of digitonin-permeabilized human umbilical vein EC with GTP gamma S, a stable GTP analog, or AIF4-, but not with GDP beta S, stimulates IP3 accumulation. However, GDP beta S inhibits GTP gamma S-induced IP3 accumulation. Although thrombin alone is not very effective in elevating IP3 levels in permeabilized EC, thrombin and GTP gamma S act in a synergistic fashion to increase IP3 accumulation. Overall, these observations are interpreted to indicate that a pertussis toxin-insensitive G protein is a key intermediate in the signaling pathway linking thrombin receptors to phospholipase C in human umbilical vein EC. PMID- 2552883 TI - Minimal inhibitory concentrations of rifabutin, ciprofloxacin, and ofloxacin against Mycobacterium tuberculosis isolated before treatment of patients in Taiwan. AB - Minimal inhibitory concentrations (MICs) of rifabutin, ciprofloxacin, and ofloxacin were determined for "wild" Mycobacterium tuberculosis strains, susceptible to all antituberculosis drugs in the conventional test, isolated from newly diagnosed Taiwanese patients who had never had prior treatment for tuberculosis. These MICs were within the limits previously reported for strains isolated in the United States. The range of MICs of rifabutin for Taiwanese strains was 0.015 to 0.125 micrograms/ml; ciprofloxacin, 0.25 to 2.0 micrograms/ml; ofloxacin, 0.5 to 2.0 micrograms/ml. On the basis of an evaluation of the highest broth-determined MICs found in this and in a previous study, we suggest that the following MICs, when determined in 7H12 broth radiometrically, should be used as breakpoints to classify the strain as "susceptible": for rifabutin, 0.125 micrograms/ml or less; for ofloxacin and ciprofloxacin, 2.0 micrograms/ml or less. PMID- 2552884 TI - [Gastroenteritis caused by enteric adenovirus serotypes 40 and 41. Preliminary study]. AB - We present the preliminary results of a prospective study of childhood gastroenteritis associated with enteric adenovirus (AE-40 and AE-41). In a period of five months we have studied 499 children with enteritis; 9 (1.8%) were positive for enteric adenoviruses (latex agglutination). The mean age of the children was 16.2 months (extreme ages 10 days and 5 years). The mean duration of diarrhoea was 2.8 days with 4-5 times daily. Respiratory symptoms were present in 44.4% of the patients and one patient was infected simultaneously with Campylobacter jejuni. All clinical syndromes were self-limiting and were treated with diet and rehydration. PMID- 2552885 TI - Hypersensitivity pneumonitis and toluene diisocyanate. PMID- 2552886 TI - Measles prophylaxis for international travel. PMID- 2552887 TI - The Cushing syndrome induced by medroxyprogesterone acetate. PMID- 2552888 TI - SV40 T-antigen as a dual oncogene: structure and function of the plasma membrane associated population. AB - SV40 T-antigen (T-ag) is localized in both the nucleus (nT-ag) and plasma membrane (pmT-ag) of cells and provides multiple functions necessary for cell transformation. The pmT-ag population is structurally very similar to the nT-ag. Transport to the cell surface is by an unknown mechanism that does not involve the secretory pathway. The disposition of T-ag in the membrane exposes both the amino and the carboxyl terminus on the exterior of the cell. Nuclear-transport defective mutants of T-ag can transform established cells in culture, but not primary cells, suggesting that non-nuclear forms of T-ag may mediate some transformation-related process(es). A non-cytolytic protein extraction technique utilizing 1-butanol solubilized from SV40-transformed cells a multimeric complex composed of pmT-ag and at least five cellular proteins ranging in size from 35,000 (35K) to 60K M. Both amino- and carboxylterminal T-ag-specific monoclonal antibodies co-precipitated T-ag and the 35-60K Mr proteins, but antibodies against the internal portion of T-ag precipitated only uncomplexed T-ag. The growth state of the cells markedly influenced the expression of the T-ag containing surface complexes; more complexes were recovered from actively dividing cells than from confluent cell cultures, and suspension cells yielded more complexes than cells on a substratum. The complex exhibited a highly dynamic association with the cell membrane, as demonstrated by pulse-chase analysis. The characteristics of growth-dependent expression and rapid turnover rate suggest a functional role for the membrane complex. The identities of the cellular proteins in the complex with pmT-ag are unknown, although one member (56K) is recognized by p53-specific monoclonal antibodies. PMID- 2552889 TI - Alteration of the program of terminal differentiation caused by oncogenes in the hemopoietic progenitor cell line 32D C13 (G). PMID- 2552890 TI - Shope fibroma: a model system to study tumorigenesis by poxviruses. PMID- 2552891 TI - Host-mediated induction of tumor heterogeneity. PMID- 2552892 TI - Genetics of endogenous murine leukemia viruses. AB - Inbred strains of mice contain in the genome 40-60 endogenous proviruses related to murine leukemia virus. To assess the genetic and pathogenic consequences of these to the host, we have developed a strategy to distinguish among the three different host-range subgroups--xenotropic, polytropic and modified polytropic- by using oligonucleotide probes specific for a polymorphic region in env. Each of these proteins detects a relatively small number of bands in a Southern blot, thus permitting us to enumerate all individual proviruses of this group. Using this approach, we have determined the distribution of different proviruses among inbred and recombinant inbred (RI) strains congenic or coisogenic for specific mutants. Using the RI results, we have been able to place over 100 proviruses on the mouse genetic map. A number of these are closely linked to well-characterized mutations, and we have been able to establish that at least one mutation, hr (hairless), was caused by a proviral insertion. If the other close linkages also prove to reflect causality, we estimate that up to 5% of recessive mutations in the mouse might be caused by insertion of proviruses of this group. Using a similar probe strategy, we have followed the evolution of murine leukemia viruses during spontaneous leukemogenesis in AKR mice. We have found that the final leukemogenic (MCF) virus is a recombinant of three different endogenous parents; an ecotropic virus, a polytropic virus that directs the gp70 region of env, and a xenotropic virus (identified as the inducible element Bxv-1) that directs the LTR. In addition to the recombinations, all such viruses also have a reduplication of the enhancer region of the LTR, compared to the endogenous parent. MCF viruses are created by these three genetic changes, which occur in a reproducible fashion and appear in the thymus between 10 and 14 weeks of age. PMID- 2552893 TI - Pathogenesis of human immunodeficiency virus infection. PMID- 2552894 TI - Review of neuroendocrine carcinomas of the larynx. AB - Neuroendocrine carcinomas of the larynx are uncommon tumors of considerable scientific interest and clinical importance. They include typical carcinoid tumors, atypical carcinoid tumors, and small cell neuroendocrine carcinomas. This paper considers these neoplasms from a personal experience and reviews the relevant medical literature. About 200 cases of neuroendocrine carcinomas of the larynx have been reported. The diagnosis is based on light microscopy and is confirmed by ultrastructural evidence of neurosecretory granules. Histochemical and immunocytochemical investigations may support it. Paraneoplastic syndromes associated with laryngeal neuroendocrine carcinomas have been reported occasionally. The histogenesis, treatment, and prognosis of these lesions also are discussed. PMID- 2552895 TI - An evaluation of the preoperative diagnosis and management of cystosarcoma phyllodes. AB - Cystosarcoma phyllodes is a rare tumour of the breast which is notoriously difficult to diagnose accurately preoperatively. In this review we report the clinical, imaging and histopathological features of 20 patients who have presented in our Centre over a 10-year period. PMID- 2552896 TI - Nucleic acid hybridization and the polymerase chain reaction in biology and medicine, with special reference to the detection of human papillomaviruses. AB - Nucleic acid hybridization, and more recently, gene amplification by the polymerase chain reaction, have contributed immensely to the development of molecular genetics. These powerful tools have been greeted and employed with great enthusiasm, in view of their wide-ranging applications in biological research and clinical diagnosis. Here we review the basic principles and some applications of these techniques, particularly in infectious diseases and oncogenesis. In order to illustrate their use, data on specific methods as applied to the detection of human papillomaviruses is presented. Gene probe and DNA amplification technology will undoubtedly continue to provide critical molecular pathways in the biomedical research endeavour. PMID- 2552897 TI - The calcium endocrine system. AB - The role of calcium as a modulator of a large number of physiological processes is well known. It is not surprising that the two principally in the understanding of its subcellular physiology and molecular biology. There is new knowledge on hepatocellular uptake of vitamin D. The subsequent hepatic conversion of vitamin D to 25 hydroxy vitamin D may indeed be under the feedback control of the active vitamin D, 1,25 dihydroxyvitamin D. Vitamin D now also appears to have many new and diverse functions some of which already have clinical application. PMID- 2552898 TI - The genomes of the human herpesviruses: contents, relationships, and evolution. PMID- 2552899 TI - Transcription and reverse transcription of retrotransposons. PMID- 2552900 TI - Nucleotide sequence of the aacC2 gene, a gentamicin resistance determinant involved in a hospital epidemic of multiply resistant members of the family Enterobacteriaceae. AB - A gentamicin resistance determinant of a conjugative plasmid from Enterobacter cloacae was cloned on a 3.2-kilobase fragment in the PstI site of pBR322. Substrate profiles for eight aminoglycosides at three concentrations showed that the resistance was due to aminoglycoside-(3)-N-acetyltransferase isoenzyme II. Insertion mapping by the gamma-delta transposon revealed that the size of the gene was approximately 1 kilobase. Nucleotide sequencing of the aacC2 gene identified an open reading frame of 858 base pairs, preceded by a promoter and a ribosome-binding site. From these data the molecular mass of the protein was calculated to be 30.6 kilodaltons. A comparison of the nucleotide sequence of the aacC2 gene with those published for the aacC3 and aacC4 genes showed complete homology of the aacC2 gene and the presumed aacC3 gene. An internal restriction fragment of the gene used as a probe in colony hybridization demonstrated the presence of the aacC2 gene in 86% of 86 multiply resistant isolates of the family Enterobacteriaceae obtained during an 18-month hospital epidemic. This corroborates our earlier data on the enzyme identification by the susceptibility profiling method. PMID- 2552901 TI - In vitro postantibiotic effect of daptomycin (LY146032) against Enterococcus faecalis and methicillin-susceptible and methicillin-resistant Staphylococcus aureus strains. AB - The suppression of bacterial growth that persists after brief exposure to antimicrobial agents has been termed the postantibiotic effect (PAE). This pharmacodynamic interaction varies for each microorganism-antimicrobial agent combination. Daptomycin (LY146032) is a new lipopeptide antibiotic with activity against gram-positive organisms. We studied the in vitro bactericidal activities and PAEs of the following drugs: daptomycin compared with penicillin G and vancomycin, without and with gentamicin against Enterococcus faecalis strains; daptomycin compared with nafcillin and vancomycin against methicillin-susceptible Staphylococcus aureus strains; and daptomycin compared with vancomycin against methicillin-resistant S. aureus strains. Daptomycin, alone and when used in combination with gentamicin, exhibited greater bactericidal activity and in general produced a longer PAE than standard effective regimens against the organism strains studied. PMID- 2552902 TI - Pharmacokinetics of rifabutin. AB - We investigated the pharmacokinetics of rifabutin in 15 male patients as part of a phase I trial of the treatment of early symptomatic human immunodeficiency virus infection. Six or more patients were studied at each of four different oral dosage levels: 300, 600, 900, and 1,200 mg/day. Twelve studies were also conducted with tracer doses of intravenous radiolabeled [14C]rifabutin. Blood and urine samples were collected for at least 72 h after the first (day 1) and last (day 28) doses of rifabutin and analyzed by high-pressure liquid chromatography. The plasma concentration data were best described by a two-compartment open model with a terminal half-life of 36 h. Rifabutin was rapidly absorbed, reaching a peak concentration about 2 to 3 h after an oral dose. Peak and trough concentrations for the 1,200-mg dose were 907 and 194 ng/ml, respectively. Total body clearance was 10 to 18 liters/h. Oral bioavailability was 12 to 20%. The drug was moderately bound to plasma proteins with a free fraction of 29 +/- 2% (mean +/- standard deviation). About 10% of an administered intravenous dose of rifabutin is excreted into the urine unchanged. Renal clearance was 1.5 +/- 0.2 liters/h. The volume of distribution was large (8 to 9 liters/kg), suggesting extensive distribution into the tissues. The area under the curve for the last dose was smaller than that of the first dose, suggesting possible induction of drug-metabolizing enzymes. PMID- 2552903 TI - Efficacy of SCH39304 in murine cryptococcosis. AB - Cryptococcal meningitis is increasing in frequency, in large part because of the advent of acquired immune deficiency syndrome. Using the murine cryptococcosis model, a new oral triazole, SCH39304, has been compared with two drugs in clinical use, fluconazole and amphotericin B. BALB/c mice (nu/nu and nu/+) were challenged intracerebrally or intranasally. Oral treatment was given daily with SCH39304 at doses of 1 to 60 mg/kg of body weight or fluconazole at doses of 1 or 5 mg/kg of body weight. Amphotericin B was given intraperitoneally three times weekly, at doses of 3 or 6 mg/kg. After intracerebral challenge, SCH39304 prolonged survival in doses as low as 1 mg/kg, a dose at which fluconazole was ineffective. At equal doses, SCH39304 consistently increased survival more than did fluconazole but not longer than did amphotericin B. SCH39304 significantly lowered colony counts in brains more than did fluconazole but no more than did amphotericin B. SCH39304 was also superior to fluconazole after intranasal challenge at equal doses. SCH39304 appears to be superior to fluconazole in mice when the drugs are given at equal doses. Clinical trials are warranted. PMID- 2552904 TI - Comparative in vitro activities of piperacillin-tazobactam and ticarcillin clavulanate. AB - The in vitro activities of ticarcillin, piperacillin, clavulanic acid, tazobactam, ticarcillin-clavulanate, and piperacillin-tazobactam against 819 bacterial isolates were compared. The two beta-lactamase inhibitors, clavulanic acid and tazobactam, had little useful antibacterial activity but enhanced the activities of the penicillins against beta-lactamase-producing strains of Haemophilus influenzae, Branhamella catarrhalis, and methicillin-susceptible Staphylococcus aureus; all strains were susceptible to both combinations. Both enzyme inhibitors also enhanced the activities of the penicillins against most strains of Escherichia coli, Klebsiella spp., Citrobacter diversus, Proteus spp., Providencia spp., and Bacteroides spp. and against occasional strains of Citrobacter freundii, Enterobacter spp., and Serratia marcescens. Clavulanic acid frequently enhanced the activity of ticarcillin against Xanthomonas maltophilia, and tazobactam frequently enhanced the activity of piperacillin against Morganella morganii. Enhancement was observed primarily with strains relatively resistant to the penicillins. In general, clavulanic acid was more effective than tazobactam in enhancing penicillin activity against Klebsiella spp., C. diversus, X. maltophilia, and Bacteroides spp., whereas tazobactam was more effective against Escherichia coli and Proteeae. There was little or no enhancement of activity against Enterococcus faecalis, Aeromonas hydrophila, Pseudomonas aeruginosa, Pseudomonas cepacia, or Acinetobacter anitratus. Clavulanic acid occasionally antagonized the activity of ticarcillin against ticarcillin susceptible members of the family Enterobacteriaceae, but those strains were still considered susceptible to the combination. Tazobactam never antagonized the activity of piperacillin. In a direct comparison of the activities of ticarcillin clavulanate and piperacillin-tazobactam, the two were equally active against H. influenzae, B. catarrhalis, and S. aureus; the latter was more active against E. faecalis. For relatively susceptible strains of members of the family Enterobacteriaceae, neither combination was predictably more active than the other, but relatively resistant strains were generally more susceptible to piperacillin-tazobactam. Piperacillin-tazobactam was more active than ticarcillin clavulanate against A. hydrophila, P. aeruginosa, and P. cepacia, similar in activity against A. anitratus, and less active against X. maltophilia and Bacteroides spp. PMID- 2552905 TI - Influence of daptomycin on staphylococcal abscesses and experimental tobramycin nephrotoxicity. AB - The antibacterial efficacies of daptomycin and vancomycin were compared in male Fischer rats with subcutaneous abscesses caused by either methicillin-susceptible Staphylococcus aureus (MSSA) or methicillin-resistant S. aureus (MRSA). The influence of daptomycin on tobramycin nephrotoxicity was also assessed. MSSA or MRSA abscesses were treated with subcutaneous daptomycin (10 mg/kg every 12 h), vancomycin (125 mg/kg every 12 h), or diluent (every 12 h) for 5 to 10 days. Rats in both antibiotic treatment groups had lower abscess bacterial counts than did controls at days 5 and 10 (P less than 0.0025). The daptomycin treatment groups had lower abscess bacterial counts than did the vancomycin treatment groups for MSSA at day 5 (P less than 0.0025) and day 10 (P less than 0.025) and for MRSA at day 10 (P less than 0.0025). Nephrotoxicity treatment groups included animals treated for 3, 7, 10, 14, and 17 days with subcutaneous diluent (every 12 h), daptomycin (20 mg/kg every 12 h), tobramycin (40 mg/kg every 12 h), and the combination of daptomycin and tobramycin. Compared with controls, animals treated with daptomycin alone exhibited no detectable nephrotoxicity. Rats given tobramycin alone developed functional and histopathologic abnormalities from days 7 through 17. Animals treated with daptomycin and tobramycin for 14 days had a lower mean concentration of creatinine in serum (P less than 0.005), higher mean creatinine clearance values (P less than 0.05), and less cortical tubular cell regeneration (P less than 0.05) than did rats treated with tobramycin alone. In rats with staphylococcal subcutaneous abscesses, daptomycin was superior to vancomycin in treating both MSSA and MRSA. Daptomycin alone caused no detectable renal injury, and in rats given daptomycin combined with tombramycin, there was less histologic and functional renal injury than in animals given tobramycin alone. PMID- 2552906 TI - Broad-spectrum antiviral activities of neplanocin A, 3-deazaneplanocin A, and their 5'-nor derivatives. AB - The neplanocin A analogs, 3-deazaneplanocin A, 9-(trans-2',trans-3' dihydroxycyclopent-4'-enyl)adenine (DHCA), and 9-(trans-2',trans-3' dihydroxycyclopent-4'-enyl)-3-deazaadenine (DHCDA), all potent inhibitors of S adenosylhomocysteine (AdoHcy) hydrolase, were studied for their broad-spectrum antiviral potential. 3-Deazaneplanocin A, DHCA, and DHCDA proved specifically effective against vesicular stomatitis virus, vaccinia virus, parainfluenza virus, reovirus, and rotavirus. Their selectivity was greater than that of neplanocin A, particularly against vesicular stomatitis virus and rotavirus. As could be expected from adenosine analogs that are directly targeted at AdoHcy hydrolase, 3-deazaneplanocin A, DHCA, and DHCDA were fully active in adenosine kinase-deficient cells, implying that their activity did not depend on phosphorylation by adenosine kinase. None of the AdoHcy hydrolase inhibitors showed selective activity against human immunodeficiency virus (type 1). 3 Deazaneplanocin A at a dose of 0.5 mg/kg per day conferred marked protection against a lethal infection of newborn mice with vesicular stomatitis virus. PMID- 2552907 TI - Mobility of gentamicin resistance genes from staphylococci isolated in the United States: identification of Tn4031, a gentamicin resistance transposon from Staphylococcus epidermidis. AB - Homologous genes encoding resistance to gentamicin, tobramycin, and kanamycin through the bifunctional acetylating [AAC(6')] and phosphorylating [APH(2")] aminoglycoside-modifying enzyme were identified in staphylococci isolated from patients in the United States. The mobility of gentamicin resistance (Gmr) genes found on a prototype conjugative plasmid (pGO1) was compared with that of genes cloned from chromosomal sites. Plasmid-encoded Gmr genes and flanking sequences were introduced onto a temperature-sensitive plasmid (pRN3208) from pGO1 by homologous recombination between insertion sequence-like elements present on both replicons. Growth of Staphylococcus aureus strains containing the temperature sensitive recombinant (pGO161) at the nonpermissive temperature for plasmid replication (42 degrees C) revealed no translocation of Gmr from its plasmid location. A transposon (Tn551) resident on the same replicon did translocate. Chromosomal Gmr determinants were cloned, together with the gene for trimethoprim resistance (dfrA), from three geographically distinct S. epidermidis isolates; two were subcloned onto temperature-sensitive Escherichia coli-S. aureus shuttle plasmids as 7.2-kilobase BglII fragments. Growth of both recombination-deficient and-proficient S. aureus strains containing the cloned genes at 42 degrees C allowed detection of transposition of Gmr sequences and identification of insertion into random chromosomal sites. We have designated this 5-kilobase transposon from S. epidermidis as Tn4031. PMID- 2552908 TI - Cloning and hybridization analysis of ermP, a macrolide-lincosamide-streptogramin B resistance determinant from Clostridium perfringens. AB - The erythromycin resistance determinant from Clostridium perfringens CP592 was cloned and shown to be expressed in Escherichia coli. The resultant plasmid, pJIR122 (7.9 kilobase pairs [kb]), was unstable since in both recA+ and recA E. coli hosts spontaneous deletion of 2.7 kb, including the erythromycin resistance determinant, was observed. Subcloning, as well as deletion analysis with BAL 31, localized the erythromycin resistance gene (ermP) to within a 1.0-kb region of pJIR122. A 0.5-kb fragment internal to ermP was 32P labeled and used as an ermP specific probe in DNA hybridization experiments which used target DNA prepared from representatives of each of the known erm classes and also from erythromycin resistant isolates of a variety of clostridial species. Hybridizing sequences were detected in DNA from several Clostridium difficile isolates and a Clostridium paraputrificum strain; however, ermP was not widespread in erythromycin-resistant C. perfringens isolates. The ermP determinant hybridized to, and shared significant restriction identity with, the ermB class gene from the streptococcal plasmid pAM beta 1. No hybridization was detected with the other six hybridization classes of erm determinants. PMID- 2552909 TI - Continuous infusion of high-dose acyclovir for serious herpesvirus infections. AB - Thirteen patients with herpesvirus infections who were unresponsive to at least 72 h of intermittent acyclovir administration received high-dose continuous infusion. Steady-state concentrations were maintained at between 20 and 98 mumol/liter. Of 12 patients who had continuous infusion for greater than 5 days, 7 (58%) resolved their infections, as determined by clinical and virologic parameters, suggesting that continuous infusion may succeed in some patients who do not respond to conventional therapy. PMID- 2552910 TI - In vitro activities of daptomycin and other antimicrobial agents against vancomycin-resistant gram-positive bacteria. AB - A comparative evaluation of daptomycin and eight other antimicrobial agents was performed by the agar dilution technique with 56 strains of vancomycin-resistant gram-positive bacteria, including Leuconostoc, Lactobacillus, and Pediococcus spp. Erythromycin, deptomycin, clindamycin, and gentamicin exhibited the greatest activities, whereas penicillin, ampicillin, and cefotaxime showed moderate activities. The organisms were all highly resistant to vancomycin and cefoxitin. PMID- 2552911 TI - In vitro studies of Chlamydia trachomatis susceptibility and resistance to rifampin and rifabutin. AB - Although rapid emergence of antibiotic-resistant mutants of Chlamydia trachomatis occurs when the organism is grown in subinhibitory concentrations of rifampin, no such mutants could be demonstrated when the organism was propagated under the same conditions in subinhibitory levels of the related drug rifabutin. PMID- 2552912 TI - Herpes simplex virus ribonucleotide reductase mutants are hypersensitive to acyclovir. AB - Two mutants defective in herpes simplex virus-encoded ribonucleotide reductase activity exhibited the novel phenotype of hypersensitivity to acyclovir, aphidicolin, and to a lesser extent, phosphonoacetic acid. These results have implications for acyclovir resistance and the development of drugs that potentiate acyclovir action by inhibition of viral ribonucleotide reductase. PMID- 2552913 TI - Effect of isoflavans and isoflavenes on the infection of Frp/3 cells by hepatitis A virus. AB - The effect of 6,4'-dichloroflavan and of isoflavan and isoflavene derivatives on hepatitis A virus (HAV) infection in a monkey cell line (Frp/3 cells) was studied. These compounds were not virucidal and had no measurable effect on the adsorption of virus to the cells at 0 degrees C, whereas they exerted an inhibitory effect on viral antigen synthesis when incubated with the infected cells during HAV multiplication. Among the substances tested, 6,4'-dichloroflavan and 6,4'-dichloroisoflavan showed the highest activity. These compounds are postulated to interact with an early stage (penetration and/or uncoating) of HAV infection. PMID- 2552914 TI - Oral chemotherapy of fatal B virus (herpesvirus simiae) infection. AB - Acyclovir and ganciclovir, which were only about 10-fold less effective against B virus than herpes simplex virus type 1 in VERO cells, were tested in vivo in B virus-infected rabbits. Untreated control rabbits became paralysed from 8 days and died from 10 days. Oral acyclovir at a dose rate of 500 mg/kg/day for 21 days prevented death; acyclovir prevented disease at 700 mg/kg/day. In B virus infected humans such a high dose of acyclovir could not be given by mouth. Nevertheless, high dose oral acyclovir is suggested for immediate prophylaxis when monkey handlers have been exposed to potentially fatal B virus infection. Should signs or symptoms of disease occur then high dose intravenous acyclovir has been recommended. Since ganciclovir was found to be more effective than acyclovir, intravenous ganciclovir might be preferred for the treatment of established infection. PMID- 2552915 TI - Inhibition of herpes simplex virus infection by tannins and related compounds. AB - Several chemically defined plant extracts were investigated for their antiviral action on herpes simplex virus (HSV-1, HSV-2)-infected African green monkey kidney cells and human adenocarcinoma cells, using a plaque formation assay. Among them, the monomeric hydrolyzable tannins, oligomeric ellagitannins and condensed tannins, having galloyl groups or hexahydroxydiphenoyl groups, had the most potent anti-HSV activity. Their 50% effective doses (0.03-0.1 microgram/ml) were by two-three orders of magnitude lower than their 50% cytotoxic doses (greater than 10 micrograms/ml). On the other hand, gallic acid, neutral polysaccharides, chemically modified (N,N-dimethylaminoethyl-, carboxymethyl-, and sulfated-) glucans, sialic acid-rich glycoproteins, and uronic acid-rich pine cone polysaccharide showed little or no activity. Using radiolabeled virus particles, we demonstrated that the anti-HSV effect of the tannins is due to inhibition of virus adsorption to the cells. PMID- 2552916 TI - Effects of inhibitors of the renin-angiotensin system on water intake after insulin administration. AB - Male wistar rats drank in a dose-related manner, in response to 1 to 40 U/kg of i.p. insulin. Maximum intakes took place during the first 30 min after i.p. insulin administration, coinciding with the period of maximal drop of blood glucose. Plasma renin activity (PRA) in rats treated with i.p. insulin was higher than in basal conditions or after saline injection. Nephrectomy and adrenalectomy did not abolish insulin-induced drinking. A low dose of captopril (0.1 mg/kg s.c.) did not modify insulin-induced drinking, but a higher dose (10 mg/kg s.c.), or enalapril (0.5 mg/microgram s.c.), significantly increased insulin-induced drinking. Enhancement of insulin-induced drinking by s.c. captopril was not secondary to an increased diuresis. Captopril (50 micrograms i.c.v.) significantly reduced the cumulative water intake after i.p. insulin (20 U/kg i.p.) plus s.c. captopril (10 mg/kg). The blockade of central receptors for angiotensin II with sarile-AII (5 micrograms) significantly diminished insulin induced drinking. It appears that the peripheral renin angiotensin system is not necessary for insulin-induced drinking but central angiotensin II plays an important role. PMID- 2552917 TI - Preparation, characterization, and application of a novel immobilized carboxypeptidase B. AB - Pig pancreas carboxypeptidase B has been immobilized by covalent attachment to a polyacrylamide-type bead support possessing carboxylic functional groups activated by water-soluble carbodiimide. The optimum conditions of immobilization were determined. The activation of the support and the coupling reaction were performed in 0.1 M sodium citrate/sodium phosphate buffer (pH 4.5) using a support-carbodiimide-enzyme weight ratio 4:8:1 at 0-4 degrees C. Under such conditions, the highest activity achieved was 6700 U/g solid. The catalytic properties and stability of immobilized carboxypeptidase B were studied and compared with the corresponding properties of the soluble enzyme. The specific activity of the immobilized enzyme calculated on bound protein basis was about 70% of that of soluble enzyme. The optimum pH for the catalytic activity of the immobilized carboxypeptidase B was practically identical with that of soluble enzyme (pH 7.6-7.7). The apparent optimum temperature of the immobilized carboxypeptidase B was about 7 degrees C higher than that of the soluble enzyme. With hippuryl-L-arginine as substrate, Kmapp value of the immobilized enzyme was tenfold higher than the Km value of the soluble enzyme. The conformational stability of the enzyme was markedly enhanced by the strongly hydrophylic microenvironment in a wide temperature and pH range. The immobilized carboxypeptidase B was used for stepwise digestion of cytochrome C. PMID- 2552918 TI - Vero cell assay for rapid detection of Clostridium perfringens enterotoxin. AB - A rapid assay which measured the biological activity of Clostridium perfringens enterotoxin was developed. The method involved the rapid killing of Vero cells by enterotoxin produced by C. perfringens grown in Duncan and Strong sporulation medium. Serial dilutions of toxin were added to Vero cells either in suspension or grown as monolayers in wells of a 96-well cell tissue culture cluster plate. Vital staining of Vero cells with neutral red, followed by extraction of the dye, allowed toxin levels to be determined either visually or by optical density measurements with a micro-ELISA M580 computer program. The toxin produced was confirmed as different from the Vero toxin of Escherichia coli and the alpha and theta toxins of C. perfringens. PMID- 2552919 TI - Biological reductive dechlorination of tetrachloroethylene and trichloroethylene to ethylene under methanogenic conditions. AB - A biological process for remediation of groundwater contaminated with tetrachloroethylene (PCE) and trichloroethylene (TCE) can only be applied if the transformation products are environmentally acceptable. Studies with enrichment cultures of PCE- and TCE-degrading microorganisms provide evidence that, under methanogenic conditions, mixed cultures are able to completely dechlorinate PCE and TCE to ethylene, a product which is environmentally acceptable. Radiotracer studies with [14C]PCE indicated that [14C]ethylene was the terminal product; significant conversion to 14CO2 or 14CH4 was not observed. The rate-limiting step in the pathway appeared to be conversion of vinyl chloride to ethylene. To sustain reductive dechlorination of PCE and TCE, it was necessary to supply an electron donor; methanol was the most effective, although hydrogen, formate, acetate, and glucose also served. Studies with the inhibitor 2 bromoethanesulfonate suggested that methanogens played a key role in the observed biotransformations of PCE and TCE. PMID- 2552920 TI - Mechanisms of microbial movement in subsurface materials. AB - The biological factors important in the penetration of Escherichia coli through anaerobic, nutrient-saturated, Ottawa sand-packed cores were studied under static conditions. In cores saturated with galactose-peptone medium, motile strains of E. coli penetrated four times faster than mutants defective only in flagellar synthesis. Motile, nonchemotactic mutants penetrated the cores faster than did the chemotactic parental strain. This, plus the fact that a chemotactic galactose mutant penetrated cores saturated with peptone medium at the same rate with or without a galactose gradient, indicates that chemotaxis may not be required for bacterial penetration through unconsolidated porous media. The effect of gas production on bacterial penetration was studied by using motile and nonmotile E. coli strains together with their respective isogenic non-gas-producing mutants. No differences were observed between the penetration rates of the two motile strains through cores saturated with peptone medium with or without galactose. However, penetration of both nonmotile strains was detected only with galactose. The nonmotile, gas-producing strain penetrated cores saturated with galactose peptone medium five to six times faster than did the nonmotile, non-gas-producing mutant, which indicates that gas production is an important mechanism for the movement of nonmotile bacteria through unconsolidated porous media. For motile strains, the penetration rate decreased with increasing galactose concentrations in the core and with decreasing inoculum sizes. Also, motile strains with the faster growth rates had faster penetration rates. These results imply that, for motile bacteria, the penetration rate is regulated by the in situ bacterial growth rate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552921 TI - Biosynthesis of 130-kilodalton mosquito larvicide in the cyanobacterium Agmenellum quadruplicatum PR-6. AB - The 130-kilodalton mosquito larvicidal gene, cloned from Bacillus thuringiensis var. israelensis, was introduced into the cyanobacterium Agmenellum quadruplicatum PR-6 by plasmid transformation. Transformed cells synthesized 130 kilodalton delta-endotoxin protein and showed mosquito larvicidal activity. Results demonstrate a potential use of a cyanobacterium for biological control of mosquitoes. PMID- 2552922 TI - Role of the membrane-associated folate binding protein (folate receptor) in methotrexate transport by human KB cells. AB - The uptake of methotrexate by KB cells was observed to be dependent on time, temperature, and concentration of extracellular methotrexate. The Kd for methotrexate surface binding to KB cells was approximately 200 nM. Following exposure of KB cells to trace quantities of [3H]methotrexate for periods ranging from 6 min to 24 h, the cellular methotrexate was progressively formed into methotrexate polyglutamates and was bound to dihydrofolate reductase as well as to a particulate folate binding protein. To further study the mechanism of methotrexate uptake in KB cells, the N-hydroxysuccinimide ester of methotrexate was used to covalently label the surface of KB cells and to inhibit transport of methotrexate. The N-hydroxysuccinimide ester of methotrexate was bound to a species of protein with an apparent molecular weight of 160,000 in 1% (v/v) Triton X-100 that bound folic acid and was specifically precipitated by antiserum raised against the previously purified high-affinity folate binding protein (the folate receptor) from human KB cells. In addition, trypsin was utilized to remove surface-accessible covalently bound methotrexate. The amount of covalently bound methotrexate that could be released by trypsin initially decreased on incubation at 37 degrees C, suggesting that the methotrexate and binding protein were internalized. However, with time, trypsin could again release the covalently bound methotrexate, suggesting that the binding protein cycles from the external cell surface to the inside of the cell and out again. PMID- 2552923 TI - Partial purification and characterization of CTP:cholinephosphate cytidylyltransferase from castor bean endosperm. AB - CTP: cholinephosphate cytidylyltransferase (EC 2.7.7.15) has been purified approximately 600-fold from postgermination endosperm of castor bean. The enzyme was solubilized with n-octyl beta-D-glucopyranoside and then subjected to ion exchange and gel filtration chromatography. The Km's of the purified enzymatic activity were 0.37 and 1.1 mM for CTP and choline phosphate, respectively. Magnesium was required for activity. The purified cytidylyltransferase activity was inhibited by both phosphate and ATP. The extent of ATP inhibition was dependent on preincubation time, temperature, and Mg2+ and Ca2+ concentrations. The possible regulation of cytidylyltransferase in castor bean endosperm by protein phosphorylation is discussed. PMID- 2552924 TI - Inhibition by linoleic acid hydroperoxide of alveolar macrophage superoxide production: effects upon mitochondrial and plasma membrane potentials. AB - Linoleic acid hydroperoxide (LOOH) is a naturally occurring product of lipid peroxidation. Incubation of rat alveolar macrophages with LOOH produced alterations of membrane properties and function at concentrations of LOOH as low as 0.1 microM. These included phorbol myristate acetate (PMA)-stimulated superoxide production, mitochondrial membrane potential, and plasma membrane potentials. These effects were clearly separated from gross loss of structural integrity as measured by lactate dehydrogenase release, in terms of both time of incubation and concentration of LOOH. PMA-stimulated superoxide production measured 15 min after addition of 10 microM LOOH was inhibited approximately 50%; however, addition of this concentration of the hydroperoxide after PMA stimulation was without effect. Superoxide production was also measured in a cell free system produced by incubation of alveolar macrophages with sodium dodecyl sulfate. Prior incubation of alveolar macrophages with LOOH, H2O2, or t-butyl hydroperoxide, under conditions that significantly inhibited superoxide production by the intact cells, did not produce inhibition of the NADPH-dependent superoxide generating system in the cell-free preparation. These results suggest that the effect of LOOH was upon signal transduction involved in the stimulation of superoxide production rather than on the NADPH oxidase itself. Measurements of membrane potential changes were made using the lipophilic ions, 3,3' dipentyloxacarbocyanine (DiOC5(3] and bis(3-phenyl-5-oxoisoxazol-4 yl)pentamethineoxonol (oxonol V). On the basis of their charge, DiOC5(3) fluorescence primarily reports mitochondrial potential and oxonol V absorbance reports plasma membrane potential. With 10 microM LOOH, depolarization of the plasma and mitochondrial membranes appeared to occur within seconds. As prior depolarization depresses superoxide production, these hydroperoxide-induced changes in membrane potential may be responsible for decreased PMA-stimulated superoxide production. PMID- 2552925 TI - Interaction of amiodarone and its analogs with calmodulin. AB - Benzofurans have important actions on the electrical properties of myocardium; the biochemical basis of those actions is not known. Crystallographic examination of these compounds has revealed that benzofurans share structural homologies with the traditional calmodulin antagonists N-(6-aminohexyl)-5-chloro-1-naphthalene and trifluoperazine. In the present study, the ability of amiodarone, desethylamiodarone, and benziodarone to displace the fluorescent ligand 8-anilino 1-naphthalene sulfonic acid (ANS) from calmodulin, to modulate the fluorescence emission of dansylcalmodulin, and to inhibit the activation by calmodulin of bovine brain cyclic nucleotide phosphodiesterase and human erythrocyte membrane Ca2+-ATPase were investigated at concentrations ranging from 10(-8) to 10(-6) M. These benzofurans displaced ANS from calmodulin with nearly equal efficiency upon forming a 1:1 complex with that protein. Each of these compounds also produced a decreased fluorescence emission of dansylcalmodulin, but with relative efficiencies being desethylamiodarone greater than amiodarone greater than benziodarone. Amiodarone and desethylamiodarone inhibited calmodulin-stimulable phosphodiesterase activity with similar potencies. Amiodarone and benziodarone inhibited calmodulin-stimulable Ca2+-ATPase activity equally, but desethylamiodarone had no effect. The observed differential effects of the amiodarone analogs suggest that calmodulin may possess multiple benzofuran binding sites that are recognized by specific targets and ligands of this Ca2+ binding protein and that the cellular action of amiodarone and its analogs may reflect calmodulin antagonism. PMID- 2552926 TI - Chondrocyte activation by interleukin-1: analysis of the synergistic properties of fibroblast growth factor and phorbol myristate acetate. AB - Following activation, monolayers of lapine articular chondrocytes secreted into their culture media large amounts of prostaglandin E2 (PGE2) and the neutral metalloproteinases collagenase and gelatinase. Partially purified preparations of synovial "chondrocyte activating factors" (CAF), which contain interleukin-1 (IL 1), generally proved stronger activators of chondrocytes than recombinant, human, IL-1 alpha (rHIL-1 alpha) or IL-1 beta (rHIL-1 beta). The presence of synergistic cytokines within the synovial material provides one possible explanation of this discrepancy. As first reported by K. Phadke (1987, Biochem. Biophys. Res. Commun. 142, 448-453) fibroblast growth factor (FGF) synergized with rHIL-1 in promoting the synthesis of neutral metalloproteinases. In our hands FGF alone did not induce neutral metalloproteinases and increased PGE2 synthesis only modestly. However, at doses from 1 ng/ml to 1 microgram/ml, FGF progressively enhanced the synthesis of PGE2, collagenase, and gelatinase by chondrocytes responding to rHIL 1. Acidic and basic FGF synergized equally well with both rHIL-1 alpha and rHIL-1 beta. Phorbol myristate acetate (PMA), but not the Ca2+-ionophore A23187, could substitute for FGF as a synergist. PMA alone was a poor inducer of collagenase or gelatinase but, unlike FGF, it greatly enhanced the synthesis of PGE2 by chondrocytes. Dot-blot analyses with a cDNA probe to collagenase mRNA confirmed that partially purified synovial CAF induced collagenase mRNA more effectively than rHIL-1, with rHIL-1 alpha being superior to rHIL-1 beta in this regard. The synergistic effects of both FGF and PMA upon IL-1-mediated collagenase induction were associated with increased abundance of collagenase mRNA. PMID- 2552927 TI - Studies on avian erythrocyte metabolism. XVII. Kinetics and transport properties of myo-inositol in chicken reticulocytes. AB - The uptake of myo-inositol was determined in a reticulocyte-enriched fraction prepared from chicken blood and compared with uptake in mature erythrocytes. While reticulocytes accumulated inositol at levels more than threefold that of the plasma concentration, erythrocyte levels were only slightly higher than that of the plasma concentration. The rate of uptake in reticulocytes was approximately 66 mumol/ml rbc/h compared to 5 mumol/ml rbc/h in mature erythrocytes when measured at an inositol medium concentration of 250 microM. The kinetic analysis of inositol influx by reticulocytes reveals a two component system: saturable and nonsaturable. The saturable component, which has a Km for inositol of approximately 222 microM, is Na-dependent. This Na-dependent saturable component, which presumably reflects active transport of inositol, accounts for 30-35% of the transport process. The saturable component is completely inhibited by amiloride but to a lesser extent by ouabain and bumetanide. Moreover, in the course of reticulocyte maturation, the saturable component is lost concomitantly with the completion of the synthesis of myo inositol pentakisphosphate and the drastic decrease in the membrane permeability to inositol. In addition, phloretin and cytochalasin B, which bind to hexose carriers and inhibit hexose sugar transport, also inhibited inositol transport. The uptake of inositol was not affected by excesses of 3-O-methylglucose (100 mM) or by physiological concentrations of D-glucose. Thus, the transport mechanism of myo-inositol appears distinct from that of D-glucose. PMID- 2552928 TI - Oxidation of the substituted catechols dihydroxyphenylalanine methyl ester and trihydroxyphenylalanine by lactoperoxidase and its compounds. AB - The reactions of native lactoperoxidase and its compound II with two substituted catechols have been investigated by ESR spin stabilization and spin trapping and by rapid scan and conventional spectrophotometric techniques. The catechols are Dopa methyl ester (dihydroxyphenylalanine methyl ester) and 6-hydroxy-Dopa (trihydroxyphenylalanine). o-Semiquinone radicals are formed in the anaerobic reaction of Dopa methyl ester with hydrogen peroxide catalyzed by native lactoperoxidase. The comparable anaerobic reaction of 6-hydroxy-Dopa appears to produce hydroxyl radicals in an unusual reaction. Compound II is reduced back to native lactoperoxidase by both catechols. The reaction between Dopa methyl ester and compound II undergoes an oscillation. The results on the overall lactoperoxidase cycle indicate two successive one-electron reductions of the peroxidase intermediates back to the native enzyme. The resulting free radical formation of o- and p-semiquinones and subsequent formation of stable quinones and Dopachromes is dependent upon the stereochemical arrangement of the catechol hydroxyl groups. PMID- 2552929 TI - Characterization of the Ah receptor for 2,3,7,8-tetrachlorodibenzo-p-dioxin: use of chemical crosslinking and a monoclonal antibody directed against a 59-kDa protein associated with steroid receptors. AB - The Ah receptor regulates induction of cytochrome P450IA1 (aryl hydrocarbon hydroxylase) by "3-methylcholanthrene-type" compounds and mediates the toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin and related halogenated aromatic hydrocarbons. Hepatic Ah receptor from untreated rodents is localized in the cytosol and has an apparent molecular mass of 250 to 300 kDa. This large form can be dissociated into a smaller ligand-binding subunit upon exposure to high ionic strength. The Ah receptor displays many structural similarities to the receptors for steroid hormones. Two non-ligand-binding proteins have been identified to be associated with the cytosolic forms of the steroid hormone receptors. The first is a 90-kDa heat shock protein (hsp 90); the second is a 59-kDa protein (p59) of unknown function. The cytosolic Ah receptor ligand-binding subunit previously has been shown to be associated with hsp 90. In the present study, we used a monoclonal antibody, KN 382/EC1, generated against the 59-kDa protein which is associated with rabbit steroid receptors to determine if p59 also is a component of the large cytosolic Ah receptor complex. Cytosolic forms of rabbit progesterone receptor, glucocorticoid receptor, and Ah receptor were analyzed by velocity sedimentation on sucrose gradients under low-ionic-strength conditions and in the presence of molybdate. Progesterone receptor from rabbit uterine cytosol and glucocorticoid receptor from rabbit liver each had a sedimentation coefficient of approximately 9 S. In the presence of KN 382/EC1 antibody the progesterone receptor and the glucocorticoid receptor both underwent a shift in sedimentation to a value of approximately 11 S. The increase in sedimentation velocity is an indication that the receptor-protein complexes are interacting with the antibody. Under low-ionic-strength conditions the Ah receptors from rabbit uterine cytosol and liver cytosol had a sedimentation coefficient of approximately 9 S. However, in contrast to the steroid receptors, the Ah receptor showed no change in its sedimentation properties in either tissue in the presence of KN 382/EC1, indicating that the antibody is not interacting with the Ah receptor. Multimeric Ah receptor complexes that were chemically crosslinked still did not show any interaction with KN 382/EC1. These data indicate that the 59-kDa protein either is not associated with the Ah receptor or is present in an altered form which the antibody cannot recognize. PMID- 2552930 TI - [Principle and therapeutic effect of lipophilic anticancer agent [SMANCS/lipiodol]: selective targeting with oily contrast medium]. AB - Lipiodol, an oily contrast medium, is utilized to deliver the anticancer agent SMANCS to the target tumor in which the tumor selective delivery of 2,500 fold more than plasma was confirmed with prolonged retention in the tumor tissue. This unique tumor targeting is accomplished by the arterial injection of the oily formulation of the drug. The method utilizes unique vascular properties of tumor tissue. SMANCS is a derivative of neocarzinostatin conjugated with copolymer of styrene and maleic acid. It has much propronounced lipophilicity, stability against various harsh environments and exerts a potent cytotoxicity. Therapeutic effect of the drug to unresectable primary hepatoma is much better than the conventional method. For Child A category patients with intrahepatic metastasis in no more than three area, a 3 yr survival rate is more than 87%. When the Child's A and B are combined with no distant metastasis, 1-, 2- and 3-year survival rates are 87%, 50%, and 35%, respectively. The side effect of this treatment [SMANCS/Lipiodol, i.p.] is minimal; transitory low grade fever is the commonest one (40-50% of cases) which can be controlled by a routine protocol. No liver or marrow toxicity was observed. Procedural limitations for the lung cancer etc. are discussed. PMID- 2552931 TI - [Basic study of chemo-embolization of the liver using hydroxyapatite granules]. AB - The purpose of this study was to elucidate the availability of hydroxyapatite (HAp) granules as a chemoembolic agent in chemo embolization therapy. A mixture of adriamycin (ADM) and an embolic agent (HAp, Lipiodol) was injected via hepatic artery in normal Wistar rats. Then the concentration of ADM in the liver serum transaminase level were measured serially. The remaining ADM in the liver was higher in groups with HAp granules than the others. The serum transaminase, however, were lower in the HAp groups. There are some advantages of HAp using as a chemo embolic agent. (1) HAp is a physiological biomaterial and seem to be safe for human. (2) HAp granules injected into the liver are easily detectable by X ray and ultrasonography. (3) HAp granules have a large surface area and this characteristic is suitable for a carrier of drugs. It is concluded that HAp granules have some necessary prerequisites for a chemo embolic agent and the application to clinical practice may be expected. PMID- 2552932 TI - [Effect of the portal vein injection of cisplatin and lipiodol on rat liver]. AB - The purpose of this study was to elucidate the efficacy and safety of cisplatin powder, suspended in lipiodol, as a means for the treatment of liver cancer by injection into the portal vein. The effects of lipiodol (0.1 ml. 0.2 ml. 0.3 ml. 0.4 ml) alone, as well as that of cisplatin powder (1.0 mg) suspended in lipiodol or saline (0.1 ml), on the liver, was investigated by administration of them into the portal vein in 100 normal Wistar rats. The results revealed that the hepatic cells were moderately affected by a small amount of lipiodol alone, but a large amount of lipiodol caused multiple anemic infarctions in the peripheral area of the liver. And, also the injection of cisplatin powder suspended in lipiodol, compared to that suspended in saline, resulted in higher concentration of platinum in the liver. Histologically severe degenerative changes were also perceived in the liver, but these changes were reversible. These results suggest that cisplatin powder suspended in lipiodol is to be used for the treatment of liver cancer. PMID- 2552933 TI - [Circulating hematopoietic stem cells in patients with small cell lung cancer]. PMID- 2552934 TI - [Slow releasing effect of adriamycin-lipiodol-emulsion made with chocola-A]. PMID- 2552935 TI - Alteration in the density, morphology, and biological properties of eosinophils produced by bullous pemphigoid blister fluid. AB - Bullous pemphigoid blister fluid (BP-BF) was examined for its effects on the density, morphology, and biological properties of eosinophils. Normodense eosinophils (NEo) were prepared from guinea pig peritoneal exudates by Nycodenz density gradient centrifugation. After culturing with BP-BF, NEo were converted into hypodense eosinophils (HEo) in a time-dependent manner. HEo were morphologically different from NEo in that HEo had spheroidal granules each with a lytic crystalloid core and a significantly increased cell volume. These HEo showed an enhanced antibody- and/or complement-dependent helminthotoxic activity to Schistosoma mansoni larvae, amplified chemiluminescence response to opsonized zymosan, and augmented expression of both FcR+ and CR+. These results suggest that BP-BF contains an activity that may not only induce an eosinophil hypodensity as a consequence of increasing cell volume, but simultaneously enhance an eosinophil cytotoxic potential through augmenting cell-surface receptors and receptor-linked oxidative metabolism. In addition, observed tissue accumulation of this activity suggests that eosinophils may be regulated by their phenotypic change in the skin lesions of bullous pemphigoid and be involved in blister formation. PMID- 2552936 TI - Detection of herpes simplex virus DNA in a cutaneous squamous cell carcinoma by in situ hybridization. AB - A case of a cutaneous squamous cell carcinoma (SCC) occurring at the site of a long-standing recurrent HSV infection is described. No deficit of the cell mediated immunity was recorded. HSV2 was isolated in several viral cultures. HSV DNA was visualized in the SCC by in situ hybridization with biotinylated probes in paraffin-embedded tissue. The samples were negative for HPV2, 5, 16, and 18 probes. A causal relationship between HSV infection and cutaneous SCC is hypothesized. PMID- 2552937 TI - [Precancerous conditions and early stages of vulvar cancer]. AB - Precancerous lesions and early stages of vulvar cancer are being increasingly observed in young women. Diagnosis is possible by means of clinical examination, colposcopy, cytology, and specific biopsy. From a prognostic and therapeutic point of view it is important to differentiate between preneoplastic and early invasive squamous cell carcinoma and to remove the lesion completely. In the case of patients with vulvar stage A carcinoma, an organ-conserving surgical procedure is justified, since a metastatic invasion of the regional lymph nodes would seem unlikely. In the case of patients with carcinoma beyond this definition an individualized tumor treatment is undertaken taking account of defined morphological prognostic criteria. PMID- 2552938 TI - [Determining indications for gynecologic supplementary treatment procedures]. PMID- 2552939 TI - [Effect of culture duration and osmolarity of the medium on nuclear surface in vitro]. PMID- 2552940 TI - [Assessment of non-palpable mammographic changes of the breast]. PMID- 2552941 TI - Still's disease associated with adenovirus infection and defect in adenovirus directed natural killing. AB - Low natural killer (NK) activity towards adenovirus infected fibroblasts was detected in the peripheral blood of a child with Still's disease and was not normalised by the addition of interferon alfa or interleukin 2. NK cytotoxicity directed at K 562 target cells or polio infected fibroblasts was normal. This specific NK deficiency might have contributed to the development of the child's Still's disease. PMID- 2552942 TI - [Estimation of the incidence of human trypanosomiasis due to Trypanosoma brucei gambiense using epidemiological parameters essentially linked to the vector: case of a focus in the Ivory Coast]. AB - The entomo-epidemiological studies carried out from 1980 to 1985 in the human trypanosomiasis focus at Bouafle (Ivory Coast) were designed to estimate the main epidemiological parameters related to G. palpalis palpalis, the sleeping sickness vector in the zone considered. This paper defines the mathematical relationships which can exist between these parameters leading to the production of a simple model enabling estimates to be made of the incidence of human trypanosomiasis caused by T.b. gambiense in this focus. The model predicted a mean annual incidence of 1.50% in the Bouafle focus. In the zones with high pig densities which were characterised by high densities of tsetse flies with weak anthropophily, the model predicted an incidence of 1.22%. However, a disease incidence of 1.76% was estimated for zones where domestic pigs were absent or at low density and characterised by low densities of tsetse flies with strong anthropophily. The author also emphasizes the relationships between mathematical models and realities in the field and the usefulness of these models to the field worker in planning control campaigns. PMID- 2552943 TI - Comparative analysis of the costs and benefits of alternative disease control strategies: vector control versus human case finding and treatment. AB - Recent estimates of the costs of detection and treatment and vector control for human trypanosomiasis are summarized and the main factors influencing their levels discussed. In order to proceed to an economic analysis a benefit unit is defined as equivalent to one year's infection avoided due to the control strategy for one person. Different monetary values could be assigned to this unit and it has the advantage of being easily integrated into epidemiological models, being represented by the difference in prevalence with and without control. A simple spreadsheet-based economic model was developed to analyse the relative economic performance of the two control strategies in terms of the cost per benefit unit obtained. The results demonstrate the sensitivity of the economic performance of each strategy to the epidemiological assumptions made, especially with respect to the existence of an animal reservoir and the evolution of the disease situation in the absence of control. They point to the need to integrate economic and epidemiological models in order to evaluate control options. PMID- 2552944 TI - Acquired immune deficiency syndrome (AIDS). Indications for abdominal surgery, pathology, and outcome. AB - Thirty-six major abdominal operations were performed on 35 Acquired Immune Deficiency Syndrome (AIDS) patients (33 men, two women). Twenty-two elective operations were indicated for diagnosis of abdominal or retroperitoneal mass (6), incomplete bowel obstruction (5), intra-abdominal infection (4), biliary symptoms (3), thrombocytopenia (3), and toxic megacolon (1). Fourteen emergency operations were for perforated viscus or peritonitis (11), massive gastrointestinal bleeding (2), and cecal volvulus (1). In 5 of 22 (23%) elective operations AIDS was unknown to the treating physicians until diagnosed by the surgical pathology; in contrast, all 14 emergency operations were in patients who had a known diagnosis of AIDS. The operative findings were related to AIDS in 34 of 36 (94%) operations. Cytomegalovirus was the most common pathogen, isolated or identified microscopically in 11 patients (eight emergency and three elective operations). Mycobacterial infections presented as retroperitoneal adenopathy or splenic abscess in six patients. Non-Hodgkins lymphoma was the most common malignancy found, presenting as an abdominal mass (4), bowel obstruction (3), or with gastrointestinal bleeding (2). Kaposi's sarcoma was diagnosed at laparotomy in four patients. The 1-month operative mortality rate for elective operation was 9% (2 of 22) and 46% (6 of 13) in emergencies. Postoperative complications included 1 reoperation for sepsis caused by inadequately resected CMV colitis; 1 pancreatic fistula; 1 wound dehiscence, and 2 minor wound infections. PMID- 2552945 TI - The clinical management of a normal contralateral breast in patients with lobular breast cancer. AB - Ninety-nine patients with the diagnosis of lobular carcinoma (LC) treated between 1970 and 1981 were reviewed. Thirteen patients had a contralateral mastectomy for duct cancer (DC) before the diagnosis of LC. Ten of the remaining 86 patients (11%) had simultaneous bilateral cancers detected by either physical examination or mammography, none by blind biopsy. Three of the surviving 38 patients (7.8%) developed a contralateral cancer an average of 143 months after operation. In comparison 167 patients with DC treated during the same period of time had a 1.8% incidence of synchronous cancer but the same incidence of subsequent cancer (7%). Lobular carcinoma in situ was not a reliable marker for predicting the presence of cancers in the contralateral breast. The diagnosis of LC is not an indication for either biopsy or removal of a normal contralateral breast. PMID- 2552946 TI - [Clinical anatomo-pathologic and surgical considerations in type III central polysyndactylia of the hand (systemization and classification of lesions)]. AB - Central polysyndactyly type III of the hands is a very rare and complicated malformation, since it shows a great number of anatomic and functional lesions. Seven persons belonging to an ethnic original group living in Trapani, in Western Sicily, have been the topic of a very deep and careful study; four of them have been operated on both sides. Our clinical research has allowed us to build up a Pedigree of the elements suffering from the malformation (showing autosomic dominant character) dating back to the beginning of the nineteenth century. The pedigree includes one hundred and fifteen persons of whom seventy healthy and forty five afflicted with central polysyndactyly type III. Numerous operative techniques have been used to face the anatomic, functional and aesthetic difficulties we have found. PMID- 2552947 TI - [Activation of the interferon system enzymes by recombinant alpha2-interferon in patients with chronic hepatitis B]. AB - Treatment of patients suffering from chronic hepatitis B with recombinant leukocytic interferon (reaferon) increased the levels of circulating interferon and activated interferon-dependent enzymes such as 2-5A-synthetase and histone kinase. Activation of the enzymes was observed for 1 to 2 weeks. After that period it was maintained at the required levels with intramuscular administration of 1-3 million units of reaferon 2 or 3 times a week. In parallel with increasing of the levels of the interferon system enzymes there was observed a decrease in the level of aminotransferase. The reaction of the viral antigens to the treatment with reaferon was not the same: HBe antigen and antibodies to HBe antigen disappeared, the content of HBs antigen and antibodies to delta interferon did not change. PMID- 2552948 TI - Fifth disease. A brief review of infections in childhood, in adulthood, and pregnancy. PMID- 2552949 TI - Itraconazole therapy for cryptococcal meningitis and cryptococcosis. AB - We studied the efficacy of itraconazole, a new oral triazole, in 33 patients (32 were immunocompromised) with cryptococcosis. Diagnoses included cryptococcal meningitis (24 patients), cryptococcemia (19 patients), cryptococcuria (4 patients), osteomyelitis (1 patient), pulmonary cryptococcosis (1 patient), and soft-tissue cryptococcosis (2 patients). Twenty-six patients had the acquired immunodeficiency syndrome, and 4 were transplant recipients. Therapy (200 mg two times per day) was monitored by clinical response, culture, and cryptococcal antigen testing. Cryptococcemia was abolished in 10 (100%) of 10 assessable patients; clinical abnormalities also cleared. Thirteen (65%) of 20 assessable patients with cryptococcal meningitis had complete responses (clinical resolution and negative cultures), 5 (25%) had partial responses, and therapy failed in 2 (10%). Ten (71%) of 14 patients with the acquired immunodeficiency syndrome who had meningitis and were treated with itraconazole as their sole therapy had complete responses, 3 (21%) had partial responses, and therapy failed in 1 (7%). Partial responses or failures were all associated with the failure of previous therapy, severe disease, low serum itraconazole concentrations, or a resistant organism. Noncompliance was associated with relapse (4 patients). Meningitis recrudesced in 3 (20%) of 15 patients who responded to therapy. All 4 patients with pulmonary cryptococcosis, soft-tissue cryptococcosis, or osteomyelitis responded to therapy (100%). Cryptococcuria was abolished in 3 (60%) of 5 assessable patients. The median survival of the 20 patients with the acquired immunodeficiency syndrome who had meningitis exceeded 10.5 months at this writing. Overall results compare favorably with amphotericin B therapy with or without flucytosine. Forty of 44 isolates of Cryptococcus neoformans were susceptible in vitro to itraconazole (minimum inhibitory concentration less than or equal to 3.13 mg/L), 3 were borderline (minimum inhibitory concentration, 6.25 mg/L), and 1 was resistant (minimum inhibitory concentration, 12.5 mg/L). As itraconazole does not penetrate cerebrospinal fluid, the meningitis results are noteworthy and suggest that meningeal and parenchymal penetration is critical. Itraconazole is promising for the treatment of cryptococcosis in patients with and without the acquired immunodeficiency syndrome. PMID- 2552950 TI - [Brain tumors associated with hemorrhage from tumors as their first manifestation]. AB - Four cases of brain tumors associated with hemorrhage from tumors as their first manifestation were reported. These were malignant astrocytoma in two cases, brain metastasis of hepatocellular carcinoma in one case and skull metastasis of hepatocellular carcinoma in one case. Clinical symptoms and sings were generalized convulsion, sudden onset of headache, vomiting and hemiparesis. It was difficult to confirm the diagnosis of brain tumors in such cases. Therefore it is important to perform follow-up plain and enhanced CT repeatedly and it is also important to make adequate biopsy of hematoma wall and surrounding tissue during operation. PMID- 2552951 TI - Usefulness of galactography for minimal noninvasive ductal carcinoma of the breast. AB - Two cases of noninvasive ductal carcinoma detected by galactography are reported with reference to our diagnostic methods of a patient with nipple discharge. Abnormal nipple discharge with no demonstrable breast lump is rare but an important clinical sign, since it is sometimes produced by malignant lesions. Non contrast mammography and cytologic examination is of limited diagnostic value for abnormal nipple discharge. Galactography is necessary for the detection of ductal carcinoma in early stage. The most important factor in improving the survival statistics for breast carcinoma is early detection. PMID- 2552952 TI - Schizotaxia revisited. AB - A conjectured neural integrative defect (schizotaxia), due to a dominant schizogene completely penetrant for a parametric aberration in synaptic signal selectivity (hypokrisia), gives rise under ordinary social learning regimens to schizotypy, a personality showing ambivalence, aversive drift, dereism, autism, and cognitive slippage. Given unfavorable polygenic potentiators (eg, introversion, hypohedonia, and anxiety) and adverse life experiences (eg, childhood trauma or adult misfortune), around 10% develop schizophrenia. That schizophrenia is basically a neurologic disorder does not contradict whatever is known about its psychodynamics, nor preclude efficacy for psychotherapy or other psychosocial interventions. Research should concentrate on soft neurology and psychophysiology as indicators, being closer in the causal chain to the schizogene than psychometric, social, or high-level cognitive processes. Taxometric statistics are appropriate to testing a major locus model not simplistically formulated. PMID- 2552953 TI - Correlation of in situ hybridization with histology and viral culture in patients with acquired immunodeficiency syndrome with cytomegalovirus colitis. AB - Endoscopic colonic biopsy specimens from 34 patients with acquired immunodeficiency syndrome and six patients without acquired immunodeficiency syndrome (3 were human immunodeficiency virus-seropositive and 3 were human immunodeficiency virus-seronegative) were examined by in situ hybridization for evidence of cytomegalovirus colitis and the results were compared with histologic examinations and viral cultures. In situ hybridization was positive in 22 of 25 patients with acquired immunodeficiency syndrome with histologic evidence of cytomegalovirus colitis. By our interpretation, 15 patients without cytomegalovirus colitis histologically all had negative hybridization studies. No correlation was found between in situ hybridization and viral culture results. In situ hybridization is a useful confirmatory test when the histologic changes are suspicious for cytomegalovirus but not considered diagnostic; it will only rarely demonstrate staining in a case considered negative histologically. PMID- 2552954 TI - Anal cancer. Microscopic condyloma and tissue demonstration of human papillomavirus capsid antigen and viral DNA. AB - Sixteen cases of squamous cell carcinoma of the anus, including 4 incidentally discovered in situ lesions, and 3 anal condylomas, were examined for the presence of human papillomavirus (HPV). All in situ tumors and 6 of the invasive tumors were associated with histologic changes typical of condyloma, despite the absence of clinical anogenital warts. Immunohistochemical studies for viral capsid antigen gave positive reactions in two anal warts and in the condylomatous area associated with one invasive tumor. In situ hybridization was accomplished using isotopic DNA probes for HPV 6/11, 16, 18, and 31. Human papillomavirus 6/11 was expressed in the corresponding capsid-positive regions in the two warts and the wart-associated invasive carcinoma. Both HPV 6/11 and HPV 16 were associated with one carcinoma in situ, and HPV 16 was also found within two invasive anal carcinomas, one of which was associated with an extensive vulvar cancer. While these observations do not resolve the "passenger" or direct oncogenic role for HPV in anal carcinoma, the circumstantial evidence is that the oncogenic influence is similar to that accepted for female genital tract cancer. PMID- 2552955 TI - Chronic cavitary respiratory papillomatosis. AB - A 19-year-old white man with multiple recurrences of respiratory papillomatosis was admitted for recurrent left lower lobe pneumonia and lung abscesses. He was found to have a single large laryngeal papilloma, widespread bronchial papillomatosis, and large cavitary lesions of the left lower lobe. A lobectomy was performed. The smooth-walled, squamous-lined cavities contained large numbers of papillomas, which were strongly positive for human papillomavirus type 11 by in situ DNA hybridization. Findings of evaluation of the patient's humoral and cell-mediated immunity were within normal limits. Cavitation appears to have resulted from bronchial obstruction, postobstructive pneumonia, and liquefactive necrosis. We speculate that squamous metaplasia allowed the continued proliferation of papillomavirus within the cavities. PMID- 2552956 TI - Etiologic studies on the association of goat pox with peste des petits ruminants (PPR) disease in Nigeria. PMID- 2552957 TI - Release of mycoplasmavirus L1 upon transfection of Acholeplasma laidlawii with homologous and heterologous viral DNA. AB - This communication reports on the release of Mycoplasmavirus L1 after infection of Acholeplasma laidlawii with purified L3 virus. Release also occurred after transfection with certain restriction fragments from MV-L3 and MV-L1 genomes. Since circular molecules are efficiently taken up in polyethylene glycol-mediated transfection, inducing fragments were applied cloned in E. coli plasmids. Release was also observed after electroporation of cells incubated with MV-L1 replicative intermediate DNA and linear MV-L3 DNA isolated from virus particles, respectively. Released MV-L1 viruses were identified after virus plaque formation on indicator lawns according to plaque morphology and hybridization with labeled viral DNA probes as well as by DNA restriction analysis. Uninfected and untransfected cells from six laboratory strains of A. laidlawii (including a MV L1 resistant one) were examined for the presence of MV-L1 DNA. They all bear MV L1 DNA integrated in their genomes. PMID- 2552958 TI - Induction by sodium butyrate of cytomegalovirus replication in human endothelial cells. AB - Cultured endothelial cells are shown to be induced in regard to permissiveness to human cytomegalovirus by temporary treatment postinfection with sodium butyrate (1-2 mM). Drug-treated cells are demonstrated to exhibit expression of immediate early and early viral antigens, synthesis of viral DNA and viral structural glycoprotein B. Progeny virus could be visualized by electron microscopy. PMID- 2552959 TI - Two different strains of an alphaherpesvirus can establish latency in the same tissue of the host animal: evidence from bovine herpesvirus 1. AB - We inoculated plaque-purified bovine herpesvirus type 1 (BHV 1), strain K22, subtype BHV 1.2b, intravenously into susceptible cattle. Five months later, we reactivated latent virus with dexamethasone and then super-infected the same cattle intranasally and intravaginally with a different plaque-purified BHV 1, strain Cooper, subtype BHV 1.1. After a second dexamethasone treatment four months later, reactivated viruses were isolated and examined with restriction endonucleases. We showed that both virus subtypes were reactivated, proving that 2 different strains of an alphaherpesvirus can establish latency in the same tissue in the host animal. PMID- 2552960 TI - The role of lipoproteins in EBV early antigen induction in Raji cells. AB - Synthesis of the Epstein-Barr virus (EBV) associated early antigen (EA) can be induced by a variety of agents in Raji cells, a latently EBV-infected Burkitt lymphoma line. We investigated the role of lipoproteins in this EA induction system. Cell growth was not affected by lipoprotein-deficiency, but EA induction by most combinations of the inducers TPA (tetradecanoyl-phorbol-acetate), IdU (iododeoxyuridine), n-BA (n-butyric acid), anti-IgM and EA inducing factor (EIF), was greatly reduced. Only the inducer combination TPA/n-BA was completely independent of the presence of lipoproteins, indicating a different induction pathway. Removing the lipid moieties of the culturing serum did not result in reduced EA induction. Thus, the lowered EA inducibility in lipoprotein-deficiency is due to the absence of the protein moiety (apolipoprotein). Addition of HDL or VLDL partially reconstituted the original EA inducibility, whereas LDL had no effect. Lipoproteins were particularly important during the first 4 hours of induction, the phase where inducers may act on cell membrane structures (e.g., receptors). But lipoproteins were also required throughout the incubation period, even in a late and inducer independent phase. PMID- 2552961 TI - Rhinovirus detection using probes from the 5' and 3' end of the genome. AB - This study investigated the abilities of cDNA probes from the 5' and 3' ends of the genome of human rhinoviruses (HRV-) 14, 9, and 1B to detect RNA from 59 rhinovirus serotypes. The results show that probes from the 5' end of the genomes of HRV-14, 9, and 1 B detected a large number of serotypes but the detection rate was variable and depended on the degree of homology with the particular probe. In contrast, all the 3' end probes were specific for the homologous virus. However, a long HRV-9 probe detected a large number of serotypes. It was concluded that such cDNA probes would not detect all serotypes with equal efficiency. Synthetic oligonucleotides corresponding to short but highly conserved regions in the 5' non coding region may overcome this problem. PMID- 2552962 TI - Immunological and molecular comparison of simian cytomegaloviruses isolated from African green monkey (Cercopithecus aethiops) and Japanese macaque (Macaca fuscata). AB - Simian cytomegaloviruses (CMV) isolated from African green monkeys and Japanese macaques were compared serologically and genetically. Twenty-three out of 31 sera from Japanese, Taiwan and rhesus macaques were positive for neutralizing antibodies to simian CMVs isolated from both African green monkeys and a Japanese macaque. The neutralizing antibody titers of those sera were slightly higher to the isolates of African green monkeys origin than to that of Japanese macaque origin. Studies with antisera prepared in guinea pigs revealed a partial antigenic cross-reaction between Japanese macaque CMV (JaM-CMV) and African green monkey CMV (AGM-CMV): the antiserum to JaM-CMV neutralized both homologous JaM CMV and heterologous AGM-CMV, but the antisera to AGM-CMV neutralized only homologous AGM-CMV. DNAs of both AGM-CMV and JaM-CMV were compared by restriction endonuclease cleavage analysis. The individual simian CMV isolates showed unique restriction patterns when cleaved with EcoRI, BamHI, or HindIII. In addition, DNA of JaM-CMV was distinguishable from that of AGM-CMV when cleaved with EcoRI or BamHI. PMID- 2552963 TI - Wall effects in sucrose density gradient centrifugation of viruses. AB - The depletion of sedimenting particles to be expected from wall effects in the course of sucrose gradient centrifugation, was computed for various types of swing-out rotors on the assumption that particles hitting the tube wall are lost from the gradient. This assumption was roughly verified for heat-denatured, but not for native poliovirus. Tween 20 reduced the depletion, and prevented tight sticking of heat-denatured virus particles to tube walls. PMID- 2552964 TI - Comparison of protective effects of serum antibody on respiratory and systemic infection of Sendai virus in mice. AB - The protective effects of the passive administration of convalescent serum from mice infected with Sendai virus were evaluated in mice challenged intranasally with wild-type and a pantropic variant (F1-R) of Sendai virus. Adoptive transfer of the serum efficiently prevented F1-R from infecting the systemic organs, but it failed to protect the mice from infections of the respiratory tracts by either virus. Virus replication in nasal turbinates was not diminished while infection in the lung was suppressed sufficiently for the infected mice to survive the infection. These findings suggest that serum antibody is less effective for the protection against viral infections on the surface of the respiratory tract, but it is effective for inhibition of spread of the virus into the systemic organs. PMID- 2552965 TI - [The role of free radical inhibitors of lipid peroxidation in protecting the myocardium from ischemic damage]. AB - The natural antioxidant beta-carotene which, unlike phenol antioxidants such as dibunol and SPN-6, is capable of exhibiting antioxidative properties under low partial oxygen pressure (ischemia), has been found to increase the activity of antioxidative enzymes in the intact and infarct myocardium and to greatly exert a more antinecrotic action when given orally in a dose of 20 mg/kg in models of rat coronary-occlusion myocardial infarction than the phenol antioxidants mentioned above. Intravenous administration of copper-containing enzymes utilizing O2 superoxide dismutase (SOD), 4 mg/kg, or ceruloplasmin, 50 mg/kg, as with a highly disperse copper powder promoting a substantial increase in antioxidative enzyme activity in the rat myocardium has been demonstrated to reduce the zone of myocardial ischemic lesion in rats and to largely enhance postoperative survival rates in the animals. Three hours following intravenous SOD, an electron microscopic examination of rat ischemic myocardium showed a considerable fall in the structural and functional damages to cardiomyocytes in the periischemic area. The findings suggest that free radical processes make a contribution to ischemic cardiomyocyte lesion and open the way for pharmacological therapy of postischemic abnormalities with enzymatic and non-enzymatic preparations of antioxidants. PMID- 2552966 TI - [Mucinous carcinoma of the breast (clinico-morphological characteristics)]. AB - The main criteria for the morphological diagnosis of the "pure" and mixed mucinous carcinoma are established on the basis of studying 83 mucinous carcinomas of the mammary gland. The necessity to differentiate is pointed out between the "pure" from of this carcinoma and the mixed one as the former has a much more favourable prognosis as compared to the latter which, in its turn, has a more favourable prognosis than an infiltrative ductal carcinoma. Total 10-year survival of patients with "pure" and mixed variants of the mucinous carcinoma is 90.46 +/- 7.27 and 58.29 +/- 11.1, respectively. The radical mammary gland resection is recommended in patients with a "pure" variant of mucinous carcinoma at 1 and 11 A stages of the disease. PMID- 2552967 TI - [Electron histochemical characteristics of laser necrosis]. AB - Linear or dot-shaped lesions were inflicted on rat liver with Nd:YAG laser, and fine structural alterations of hepatocytes were studied in the specimens processed for an endoplasmic reticulum (ER) marker glucose-6-phosphatase (GP). 5 7 s after irradiation a severe cell damage and GP inhibition occurred near the lesions, with less injured cells located laterally. 24 hr later the zone of the necrosis increased markedly. An autolytic decomposition in the newly formed necrotic area was much more pronounced as compared to the area of the initial necrosis. Phagocytic resorption of the intensively irradiated tissue was retarded this explaining some clinical observations on the long-term healing after Nd:YAG laser surgery. Based on our observations the so-called contact regimen of the irradiation is recommended due to the small size of the initial necrosis produced with this method. The various patterns of cell injuries including some changes in ER and enzyme GP as its marker are described in detail. PMID- 2552968 TI - MK-801 prevents hippocampal neurodegeneration in neonatal hypoxic-ischemic rats. AB - In cerebral asphyxia, enhanced postsynaptic stimulation of N-methyl-D-aspartate (NMDA) receptor by excessive glutamate may mediate neuronal injury and death. The neuroprotective potential of the novel, potent NMDA receptor antagonist MK-801 was assessed by evaluating hippocampal behavioral and histologic outcomes in an experimental rat model of neonatal hypoxia/ischemia. Seven-day-old rats with and without MK-801 pretreatment were subjected to unilateral carotid ligation followed by 2 hours of hypoxia. At age 30 days, spontaneous alternation behavior was measured using a conventional wooden T maze. Hypoxic-ischemic animals pretreated with saline demonstrated a significant impairment in spontaneous alternation behavior compared with that of normal control rats and the hypoxic ischemic rats pretreated with MK-801. Hippocampal neuronal damage in the CA1 and CA3 regions was prevented in animals pretreated with MK-801 vs saline-treated controls. Therefore, while saline-treated rats with hippocampal lesions showed defective memory and hippocampal neuronal destruction, pretreatment with MK-801 protected rats. Thus, MK-801 appears to protect hippocampal neurons from hypoxia/ischemia and may be potentially beneficial in preventing neonatal asphyxial brain damage. PMID- 2552969 TI - Focal cerebral infarctions associated with perivascular tumor infiltrates in carcinomatous leptomeningeal metastases. AB - Diffuse carcinomatous leptomeningeal metastases "carcinomatous meningitis") have the usual clinical course involving multifocal nerve root deficits and a variable diffuse encephalopathy. In contrast, we describe a patient with carcinomatous leptomeningeal metastases who presented with clinical signs of meningitis and focal cerebral infarction. Over an 8-month period, multiple cerebral infarctions and cranial neuropathies developed. Postmortem examination of the patient's brain revealed diffuse leptomeningeal infiltration by a signet-ring adenocarcinoma. The extensive involvement of the subarachnoid space with tumor was associated with dense neoplastic infiltration of the Virchow-Robin spaces. These perivascular tumor infiltrates were accompanied by multifocal mural invasion and, less frequently, by intravascular tumor cells obliterating the lumen. Focal hemorrhagic infarcts in the cerebral cortex corresponded to areas of microscopic vasculopathy. This case provides evidence that tumor-associated vasculopathy with resultant ischemia plays a role in the pathogenesis of focal cerebral infarctions in carcinomatous leptomeningeal metastases. PMID- 2552970 TI - Ketamine and retinal ischemia. PMID- 2552971 TI - Fibrous histiocytoma. An uncommon eyelid lesion. PMID- 2552972 TI - The human nasopharynx as a reservoir for Epstein-Barr virus. AB - Nineteen human nasopharyngeal tissue specimens biopsied from patients with suspected nasopharyngeal carcinoma (NPC) and 15 adenoid tissue specimens obtained by surgery were successfully grown in culture without bacterial and fungal contamination. Fibroblastic growth developed in all of the cultures. In some cases, epithelial growth was also seen among the fibroblastic growth. The cells became round and floated in the culture fluid between 19th and 57th day of culture in 6 out of 10 NPC tissue cultures, 3 out of 9 tissue cultures of apparently normal nasopharyngeal mucosa of patients referred for exclusion of NPC, and 4 out of 15 adenoid tissue cultures. The floating cells consisted of B lymphoblastoid cells and were positive Epstein-Barr virus (EBV)-associated nuclear antigen (EBNA). The data suggest that a small number of B-lymphocytes in the human nasopharynx serve as a reservoir for EBV. PMID- 2552973 TI - Anaplastic small cell carcinoma of the larynx. AB - Anaplastic small cell carcinoma of the larynx is an uncommon neuroendocrine tumor. We report a case of this neoplasm of a 53-year-old male. The patient showed persistent hyponatremia accompanied with continuous loss of sodium in the urine, which resulted from inappropriate secretion of antidiuretic hormone (ADH). It is postulated that hyponatremia in this case was due to production of ADH by the tumor cells. PMID- 2552974 TI - Gestational trophoblastic disease in New Zealand, 1980-1986. AB - During the period 1980 to 1986, inclusive, 350 cases of gestational trophoblastic disease were recorded within New Zealand; of these cases nearly 70% were reported to a Register established to obtain epidemiological information. Clinical information obtained with the notifications revealed no difference in incidence of gestational trophoblastic disease among the 3 main ethnic groups which make up the New Zealand population; the uterine fundus was recorded as being large for dates in only 26%; the most common clinical presentation was as a threatened abortion; 7% of the cases were diagnosed at either routine ultrasound examination or at termination of pregnancy, there having been no suspicions prior to that procedure. A review of histological material obtained following notification suggested that the histological diagnosis of trophoblastic disease could not be substantiated in 14.7% of cases. During the period under review, therefore, there were 299 cases of trophoblastic disease (all but 2 of which were hydatidiform mole) and 447,667 pregnancies giving an incidence of 1 case of trophoblastic disease per 1,497 pregnancies. PMID- 2552975 TI - Use of alpha-2-interferon nasal spray for prophylactic reduction of rhinovirus induced infection in mission-critical groups. PMID- 2552976 TI - Post-training systemic and intra-amygdala administration of the GABA-B agonist baclofen impairs retention. AB - The effects of the GABA-B receptor agonist baclofen on memory storage were studied in two series of experiments. In the first series, CD-1 mice were trained in two aversively motivated tasks: a one-trial inhibitory avoidance task and a classical conditioning task (conditional emotional response). Immediate post training ip administration of (+/-)baclofen (10 and 30 mg/kg) impaired retention of animals in both tasks. The effect was time-dependent: Retention was not affected by baclofen administered 120 min after training. In the second series of experiments, which used Sprague-Dawley rats, post-training intra-amygdala administration of baclofen impaired retention of an inhibitory avoidance response. These results support the view that the GABAergic system is involved in the modulation of memory storage and that the amygdaloid complex may be a critical site for effects of drugs affecting the GABAergic system. PMID- 2552977 TI - Effects of ketamine on tunnel maze and water maze performance in the rat. AB - The NMDA receptor, which has been implicated in memory formation, is noncompetitively blocked by ketamine. The present study examines the effect of ketamine (0, 3, 6, 12, and 25 mg/kg body wt; ip) on tunnel maze and water maze performance in Wistar rats. In the hexagonal tunnel maze (HTM) high doses of ketamine (12 and 25 mg/kg) decreased locomotor activity. Moreover, ketamine induced perimeter walking (6, 12, and 25 mg/kg) and attenuated exploratory efficiency (25 mg/kg). When the HTM was converted into a modified six-arm radial maze, ketamine impaired short-term but not long-term memory. In the Morris water maze, rats injected with ketamine (12 and 25 mg/kg) acquired a spatial navigation task more slowly than controls. When the escape platform was removed, the drug treated rats did not preferentially search for it in the area where the platform had been during the acquisition phase. However, when the escape platform was visible, no differences in the performance of ketamine-treated and control rats could be found. In summary, ketamine seems to attenuate some but not all forms of learning in the tunnel maze and it impairs the acquisition of a spatial navigation task. PMID- 2552978 TI - ACTH 4-9 analog can retard spatial alternation learning in brain damaged and normal rats. AB - Thirty adult male hooded rats (Long-Evans strain) were assigned randomly to one of three lesion groups (n = 10) and prepared with medial frontal, posterior parietal, or sham neocortical injuries. Following a recovery interval of 10-12 days, access to water was limited to 30 min per day and the rats were shaped to traverse a T-maze for a reward of sweetened water. After a pretraining criterion was attained, osmotic minipumps (Alzet 2002) were installed subcutaneously. The minipumps delivered chronically for the next 14 to 15 days either 0 or 1.2 micrograms of ACTH 4-9 dissolved in bacteriostatic saline per day while the rats were trained on a reinforced spatial alternation task. Analysis of the number of errors made to a criterion of at least 80% correct alternations in two consecutive training sessions, or a ceiling of 62 errors (attained by two rats with parietal lesions), revealed that learning was impaired in the rats with parietal injuries. Contrary to our hypothesis, animals receiving ACTH 4-9 committed more errors than their counterparts receiving only saline. PMID- 2552979 TI - Models for C1. Tools or toys? The real biological challenge. AB - C1 modelling, based on structural and functional data, does not yet bring the different laboratories to a consensus on C1 activation, activity and associated controls. The heart of C1 beats in its subcomponent C1r2, which, from its domain structure and its twinning with subcomponent C1s, represents the challenge for the knowledge of C1. The 8-shaped model proposed for the C1r2-C1s2 association, with a head-to-tail interaction between the C1r catalytic domains, appears as the hub of an active world in the bosom of C1q. More detail is now required on protein-protein interactions inside C1 to refine the available models or to propose alternatives. Precise data on the interactions of C1 proteins with activators, substrates or control proteins are also likely to bring pertinent help in proposing future models for C1. PMID- 2552980 TI - Models for the C1 complex determined by physical techniques. AB - The C1 complex is an association of C1q and C1r2C1s2. Neutron scattering and ultracentrifugation provide a valuable means of understanding the solution structure of the subcomponents and their complex, and these can be supplemented by protein structure prediction techniques. C1q is constructed from six globular heads connected by collagen-like arms. The solution data for C1q show that the arms are of length 14.5 nm and not 11.5 nm as proposed from electron microscopy, the average arm-axis angle is 40 degrees, and that the structure is flexible in solution at the junction of the six arms. The sequences of C1r and C1s show that each is constructed from six protein domains. C1r and C1s are elongated macromolecules of lengths 18-20 nm. Their solution properties are best described as the lengthwise arrangement of a protease domain of diameter 4 nm, two "short consensus repeat" domains, each of length 4 nm, and an N-terminal globular entity of length 6 nm containing the first three protein domains. Solution data on the C1r2 dimer is interpreted as an X-shaped association of the two C1r monomers as proposed from electron microscopy. Six criteria are enumerated for constructing models of C1 from these two structures, and four distinct models for the C1 complex are reviewed. While further evidence is required to make this choice unequivocal, the W-model is favoured. This places each monomer of C1r and C1s on four adjacent arms of C1q, and offers the most reasonable explanation of the known properties of the C1 complex. PMID- 2552981 TI - Interaction of fluid phase C1/C1q and macrophage membrane-associated C1q with gram-negative bacteria. AB - Many gram-negative bacteria are killed after treatment with normal non-immune sera and directly bind and activate C1 in the absence of antibodies. For the immediate killing of such serum-sensitive bacteria, like R-forms of Salmonella strains, all serum complement components are essential. When purified serum C1 to C9 are used, further activation of the cascade requires an additional serum factor. This glycoprotein differs from antibody and mediates the attachment of C4b to the bacterial cell surface. The antibody-independent interaction with C1 occurs via C1q, which binds to LPS. In addition outer membrane proteins bind C1q and C1. The association of these porins with LPS may potentiate the antibody independent C1q and C1 binding to serum-sensitive bacteria. Porins can contribute to complement activation mainly through the classical pathway. LPS and porins from bacterial cell walls are also involved in the binding of gram-negative bacteria to macrophages. This antibody-independent attachment and ingestion of gram-negative bacteria is mediated by endogenous macrophage-membrane associated C1q. PMID- 2552982 TI - Structure and function of C1r and C1s: current concepts. AB - C1r and C1s, the constituent proteins of C1s-C1r-C1r-C1s, the Ca2+ -dependent catalytic unit of C1, are homologous serine proteinases that share a common activation pattern and have similar structural organizations at the monomeric level. In both cases, activation occurs through cleavage of a single Arg-Ile bond, which converts the single-chain proenzymes into active proteinases comprising two chains linked by a single disulphide bridge. Both NH2-terminal A chains are sub-divided into five structural units (I-V) including a single copy of an Epidermal Growth Factor-like segment (II) and two different pairs of internal repeats (I/III and IV/V). Regions I and III have no equivalent in other proteins, whereas regions IV and V are homologous to short consensus repeats found, in particular, in complement proteins C2, B, H, C4b-binding protein and CR1. The COOH-terminal B chains are homologous to the catalytic chains of serine proteinases, but lack the "histidine-loop", a disulphide bridge common to all other known mammalian serine proteinases. Overall sequence comparison of C1r and C1s reveals 40% amino acid identity and conservation of all cysteine residues. In contrast, C1r and C1s widely differ from each other by their glycosylation patterns: both proteins contain Asn-linked carbohydrates, but four glycosylation sites are present on C1r, and only two on C1s.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2552983 TI - The structures of human C1r and C1s and their relationship to other serine proteases. AB - The recent sequencing of the C1 subcomponents has allowed comparison with other molecules of homologous primary structure. Where tertiary structures are available for at least one member of the family it is possible to make further progress by modelling the amino acid sequence of the complement protein into the three-dimensional coordinates of the directly determined structure, thereby obtaining an approximation of the structure of the complement protein. Molecular modelling allows structure-function relationships to be explored and suggests further experiments that may be amenable to techniques such as site-directed mutagenesis. PMID- 2552984 TI - Biosynthesis of C1r and C1s subcomponents. AB - Biosynthesis of C1r and C1s subcomponents has been studied using monocytes and macrophages, hepatocytes and hepatoma cell lines or fibroblasts. Both proteins have been detected in supernatants and cell lysates as proenzymic monocatenar molecules. C1r and C1s were secreted by stimulated monocytes and by Hep G2 cells, according to a 1:1 stoichiometry. Monocyte C1s secretion was enhanced by lymphokines, such as alpha- or gamma-interferon or by placental soluble factors. Expression of both proteins was coordinately modulated by a newly purified 14 kDa lymphokine at a pretranslational level. Data from in vitro RNA translation are discussed. PMID- 2552985 TI - [Semi-malignant giant cell synovioma as an incidental finding in open dislocated fracture of the upper ankle joint]. AB - A rare statement is demonstrated with reference to a case concerning a sixteen years old girl who caught an open luxation fracture of her upper ankle joint by a sudden fall from a horse on August 8, 1988. During the operative treatment a spontaneously loosing soft tissue tumor of plumlike size could be obtained. The histological analysis proved a semimalignant giant cell-synovialom. PMID- 2552986 TI - [The seasonal toxigenicity of different cultured plants for clostridia in relation to so-called wildlife mortality]. AB - In the juice of plants which could be eaten by hares different amounts of toxins (haemolysin, lecithinase) could be found after the partly addition of a C. perfringens field strain and subsequent anaerobic incubation. Sterile filtrates showed a very pronounced toxigenicity. The presented results proof in tendency that oilseed-rape (00-rape seed), wheat, and barley as green plants can contribute in clostridial toxicosis in hares, whereas grass and beets are involved only partially, and clover is practically completely atoxigenic. PMID- 2552987 TI - Polyamines and neomycin inhibit the purified plasma-membrane Ca2+ pump by interacting with associated polyphosphoinositides. AB - We investigated the effect of spermine, spermidine, putrescine and neomycin on the activity of the plasma-membrane Ca2+ pump and on its stimulation by negatively charged phospholipids and calmodulin. Millimolar concentrations of spermine and to a lesser extent of spermidine decreased the ATPase activity in the presence of phosphatidylinositol 4,5-bisphosphate (PIP2), without affecting the stimulation by phosphatidylinositol 4-phosphate (PIP). Sub-millimolar concentrations of neomycin inhibited the stimulation of the ATPase by PIP and by PIP2. Neomycin was more effective at the higher concentrations of PIP and PIP2. We discuss that these findings are compatible with the hypothesis that PIP and PIP2 bind to the ATPase and that several of these molecules have to be available to stimulate the ATPase. PMID- 2552988 TI - Inhibition of inositol 1,4,5-trisphosphate 3-kinase by heparin: basal and Ca2+/calmodulin-stimulated activity. PMID- 2552989 TI - Effect of propan-2-ol on enzymic and structural properties of elongation factor G. AB - Elongation factor G (EF-G) can support a GTPase activity in vitro even in the absence of ribosomes when propan-2-ol is present [GTPasep; De Vendittis, Masullo & Bocchini (1986) J. Biol. Chem. 261, 4445-4450]. In the present work the GTPasep activity of EF-G was further studied by investigating (i) the effect of ionic environment on GTPasep and (ii) the influence of propan-2-ol on the molecular structure of EF-G as determined by fluorescence and c.d. measurements. In the presence of 1-300 mM univalent cations (M+) alone, no detectable GTPasep activity was measured; however, in the presence of 1 mM-Mg2+ a considerable stimulation was observed at 40 mM-Li+ or 75 mM-NH4+. Among bivalent cations (M2+), 1 mM-Sr2+, 2-5 mM-Ca2+ and 1 mM-Ba2+ were the most effective, but, in the presence of 75 mM NH4+, Mg2+ and Mn2+ became the most efficient, whereas the stimulation by other M2+ species was considerably decreased. C.d. measurements showed that the alcohol increased the mean molar residue ellipticity of EF-G at 285 nm, but not at 220 nm. As estimated from fluorescence measurements, in the presence of 20% (v/v) propan-2-ol the value of the dissociation constant of the complex formed between EF-G and 8-anilino-1-naphthalene-sulphonate decreased from 8 to 5 microM; similarly, the number of binding sites on EF-G for the fluorescent probe decreased from 13 to 6. Finally, the alcohol enhanced the quenching of the intrinsic fluorescence of EF-G caused by either acrylamide or KI. The data support the hypothesis that propan-2-ol induces moderate conformational changes of EF-G that make the catalytic centre accessible to the substrate even in the absence of ribosomes. Kinetics of GTPasep studied at different temperatures did not reveal additional structural changes of EF-G occurring with time or temperature. PMID- 2552990 TI - The rate of the AMP/adenosine substrate cycle in concanavalin-A-stimulated rat lymphocytes. AB - The effect of adenosine on the metabolism of prelabelled adenine nucleotides was investigated in concanavalin-A-stimulated rat lymphocytes. Adenosine in the presence of the adenosine deaminase inhibitor, deoxycoformycin, caused a 2-fold increase in the ATP concentration. This effect was, in part, countereacted by an increased rate of adenine nucleotide catabolism, which could be explained by a stimulation of AMP deaminase (EC 3.5.4.6). At the same time a continuous rate of labelled adenosine production was found, which was not affected by the increased ATP concentration and which could only be detected by the trapping effect of a high concentration of added unlabelled adenosine. It is concluded that the rate of the substrate cycle between AMP and adenosine is low (1.9 +/- 0.2 nmol/h per 10(7) cells) in comparison to the rate of AMP deamination. PMID- 2552991 TI - Activation of electropermeabilized neutrophils by adenosine 5'-[gamma thio]triphosphate (ATP[S]). Role of phosphatases in stimulus-response coupling. AB - Electrically permeabilized human neutrophils were used to study the mechanism of activation of the NADPH oxidase by chemotactic factors. The respiratory burst elicited by formyl-methionyl-leucyl-phenylalanine (fMLP) was strictly dependent on the addition of ATP. The response was also supported by adenosine 5'-[gamma thio]triphosphate (ATP[S]), but not by the non-hydrolysable analogue (p[NH]ppA). In the presence of ATP, displacement of fMLP from its receptor by antagonist peptides resulted in the abrupt termination of the O2-consumption burst. In contrast, the response persisted after displacement of fMLP when ATP[S] was present. This finding is consistent with the formation of biologically active thiophosphoproteins which are resistant to cleavage by cellular phosphatases. Accordingly, lower concentrations of ATP[S], as compared with ATP, were required to support the fMLP response. The data indicate that protein phosphatases control the extent and duration of the response in cells stimulated with chemoattractants. Unlike ATP, sub-millimolar concentrations of ATP[S] elicited a spontaneous respiratory burst in the absence of fMLP or other stimuli. This effect was inhibited by p[NH]ppA and was not observed in intact (non permeabilized) cells, indicating interaction of ATP[S] with an intracellular adenine-nucleotide-binding site, possibly a protein kinase. These results suggest that protein kinases are active in neutrophils in the absence of exogenous stimuli, but that accumulation of the essential phosphoprotein(s) is normally prevented by the ongoing vigorous phosphatase activity. It is conceivable that control of the respiratory burst is exerted by inhibition of phosphatase activity, instead of or in addition to the more commonly postulated activation of protein kinases. PMID- 2552992 TI - The spin trapping of pyrimidine nucleotide free radicals in a Fenton system. AB - The reaction of the hydroxyl radical, generated by a Fenton system, with pyrimidine deoxyribonucleotides was investigated by using the e.s.r. technique of spin trapping. The spin trap t-nitrosobutane was employed to trap secondary radicals formed by the reaction of the hydroxyl radical with these nucleotides. The results presented here show that hydroxyl-radical attack on thymidine, 2 deoxycytidine 5-monophosphate and 2-deoxyuridine 5-monophosphate produced nucleotide-derived free radicals. The results indicate that .OH radical attack occurs predominantly at the carbon-carbon double bond of the pyrimidine base. The e.s.r. studies showed a good correlation with previous results obtained by authors who used x- or gamma-ray irradiation to generate the hydroxyl radical. A thiobarbituric acid assay was also used to monitor the damage produced to the nucleotides by the Fenton system. These results showed qualitative agreement with the spin-trapping studies. PMID- 2552993 TI - Antibodies to junctional sarcoplasmic reticulum proteins: probes for the Ca2+ release channel. AB - The junctional face membrane plays a key role in excitation-contraction coupling in skeletal muscle. A protein of 350 kDa, tentatively identified as a component of the junctional feet, connects transverse tubules to terminal cisternae of sarcoplasmic reticulum [Kawamoto, Brunschwig, Kim & Caswell (1986) J. Cell Biol. 103, 1405-1414]. The membrane topology and protein composition of sarcoplasmic reticulum Ca2+-release channels of rabbit skeletal muscle were investigated using an immunological approach, with anti-(junctional face membrane) and anti-(350 kDa protein) polyclonal antibodies. Upon preincubation of the terminal cisternae with anti-(junctional face membrane) antibodies, Ca2+-ATPase and Ca2+-loading activities were not affected, whereas anti-(350 kDa protein) antibodies stimulated Ca2+-ATPase activity by 25% and inhibited Ca2+-loading activity by 50% (at an antibody/terminal cisternae protein ratio of 1:1). Specific photolabelling of terminal cisternae proteins with [14C]doxorubicin was prevented by both anti (junctional face membrane) and anti-(350 kDa protein) antibodies. Stimulation of Ca2+ release by doxorubicin was prevented by both anti-(junctional face membrane) and anti-(350 kDa protein) antibodies. Half-maximal inhibition was obtained at an antibody/terminal cisternae protein ratio of 1:1. Kinetic measurements of Ca2+ release indicated that anti-(350 kDa protein) antibodies prevented Ca2+-induced Ca2+ release, whereas the ATP-stimulation and the inhibition by Mg2+ were not affected. These results suggest that: (i) Ca2+- and doxorubicin-induced Ca2+ release is mediated by Ca2+ channels which are selectively localized in the junctional face membrane; (ii) the 350 kDa protein is a component of the Ca2+ release channel in native terminal cisternae vesicles; and (iii) the Ca2+ activating site of the channel is separate from other allosteric sites. PMID- 2552994 TI - The phorbol ester TPA stimulates the expression of functional beta-adrenoceptors in human T lymphoblasts Molt 3. AB - In crude membranes from human T lymphoblasts Molt 3 cultured under standard conditions, the adenylate cyclase system was stimulated by GTP, its beta gamma imido analogue (p[NH]ppG,) NaF and forskolin, but not by isoprenaline, prostaglandin E1 and vasoactive intestinal peptide. TPA (tumour-promoting agent phorbol ester) added at low concentration (3.2 nM) to the culture medium induced a marked increase in functional beta 2-adrenoceptors. Competition curves of [125I]cyanopindolol with the antagonist ICI 118.551 and four beta-adrenergic agonists indicated that the emergence of functional beta 2-adrenoceptors corresponded to one class of binding sites, shifting from a high-affinity state for agonists to a low-affinity state in the presence of p[NH]ppG. This expression of beta 2-adrenoceptors after a 4 h lag period depended on newly formed mRNA and protein synthesis as judged by the inhibitory effects of actinomycin D and cycloheximide. Further effects of TPA included alterations of the stimulatory G protein Gs and/or the catalytic unit of adenylate cyclase. PMID- 2552995 TI - Short-term regulation of glycolysis by vasoactive intestinal peptide in epithelial cells isolated from rat small intestine. AB - In epithelial cells isolated from rat small intestine, we have studied the influence of vasoactive intestinal peptide (VIP), a neurotransmitter which markedly increases enterocyte cyclic AMP, and of two cyclic AMP analogues (8 bromo cyclic AMP and N6,2'-O-dibutyryl cyclic AMP) on the rate of glycolysis, fructose 2,6-bisphosphate concentration and 6-phosphofructo-2-kinase activity, as well as on the rate of 3-O-methyl-D-[14C]glucose uptake. Our results show that, without affecting the rate of 3-O-methyl-D-[14C]glucose accumulation, VIP and cyclic AMP analogues were able to inhibit glucose consumption and L-lactate formation by isolated rat enterocytes. These effects occurred parallel to a significant decrease in the cellular concentration of fructose 2,6-bisphosphate and to a partial inactivation of 6-phosphofructo-2-kinase. These findings support the hypothesis that VIP inhibits glycolysis in rat enterocytes through a cyclic AMP-dependent mechanism. PMID- 2552996 TI - Glucagon, cyclic AMP and adrenaline stimulate the degradation of low-density lipoprotein by cultured rat hepatocytes. AB - Rat hepatocytes were preincubated for 16 h with hormones or drugs and then for a further 8 h with 125I-human low-density lipoprotein (LDL). Glucagon (via cyclic AMP) and adrenaline (via cyclic AMP and alpha-effects) increased the binding of 125I-LDL to the LDL receptor, and the degradation of LDL to [125I]iodotyrosine. The effects on degradation were antagonized by dexamethasone, and the action of cyclic AMP on binding and degradation was inhibited by actinomycin D. The results are discussed in relation to the control of lipoprotein metabolism in diabetes. PMID- 2552997 TI - Complexing of the CD-3 subunit by a monoclonal antibody activates a microtubule associated protein 2 (MAP-2) serine kinase in Jurkat cells. AB - Treatment of Jurkat T-cells with anti-CD-3 monoclonal antibodies resulted in the rapid and transient activation of a serine kinase which utilized the microtubule associated protein, MAP-2, as a substrate in vitro. The kinase was also activated on treatment of Jurkat cells with phytohaemagglutinin, but with a different time course. The activation of the MAP-2 kinase by anti-CD-3 antibodies was dose dependent, with maximal activity observed at concentrations of greater than 500 ng/ml. Normal human E-rosette-positive T-cells also exhibited induction of MAP-2 kinase activity during anti-CD-3 treatment. The enzyme was optimally active in the presence of 2 mM-Mn2+; lower levels of activity were observed with Mg2+, even at concentrations up to 20 mM. The kinase was partially purified by passage over DE-52 Sephacel with the activity eluting as a single peak at 0.25 M-NaCl. The molecular mass was estimated to be 45 kDa by gel filtration. The activation of the MAP-2 kinase was probably due to phosphorylation of this enzyme as treatment with alkaline phosphatase diminished its activity. These data demonstrate that the stimulation of T-cells through the CD-3 complex results in the activation of a novel serine kinase which may be critically involved in signal transduction in these cells. PMID- 2552998 TI - Temporal aspects of O-glycosylation of glycoprotein C from herpes simplex virus type-1. AB - Herpes simplex virus type-1 glycoprotein C (gC1) contains several O-linked oligosaccharides clustered near N-linked chains, and Pronase digestion produces glycopeptides carrying both oligosaccharide types. We have taken advantage of this fact to investigate the temporal relationship between the initiation of O linked chains and the processing of N-linked oligosaccharides. gC1 was isolated from herpes-simplex-virus-infected BHK (baby-hamster kidney) cells after short labelling periods with [3H]glucosamine, and the labelled Pronase-cleaved glycopeptides fractionated on concanavalin A-Sepharose. N [3H]Acetylgalactosamine, mostly convertible into free N [3H]acetylgalactosaminitol on mild alkaline-borohydride treatment, was found in glycopeptides with an affinity to concanavalin A-Sepharose corresponding to that of glycopeptides carrying Man8GlcNAc2 or larger N-linked chains. Since there is evidence that the processing of N-linked chains up to Man8GlcNAc2 involves enzymes located in the rough endoplasmic reticulum, current results strongly suggest that gC1 acquires O-linked N-acetylgalactosamine before the glycoprotein routing to the Golgi apparatus. The addition of the second sugar to the nascent O linked chain appeared to occur after a relatively long lag time. PMID- 2552999 TI - Stimulation of intracellular topoisomerase I activity by vasopressin and thrombin. Differential regulation by pertussis toxin. AB - Incubation of cultured rat aortic smooth muscle cells (A-10, ATCC CRL 1476) with [8-arginine]vasopressin (AVP) or thrombin increased the amount of DNA strand breakage induced by camptothecin, an inhibitor of topoisomerase I (DNA topoisomerase; EC 5.99.1.2) and transiently stimulated the extractable activity of this enzyme. Both topoisomerase-related responses were prevented by treatment of the cells with AVP or thrombin plus the appropriate receptor antagonist. The increase in strand breakage mediated by AVP and thrombin depended on the concentration of hormone. Neither AVP nor thrombin had any effect on strand breaks obtained with the epipodophyllotoxin VM-26, an inhibitor of topoisomerase II [DNA topoisomerase (ATP-hydrolysing); EC 5.99.1.3]. Pretreatment of the cells with pertussis toxin partially inhibited thrombin-mediated increases in camptothecin-induced strand breakage whereas AVP-mediated increases were unaffected. These results are consistent with the notion that AVP and thrombin induce a transient increase in intracellular topoisomerase I activity via interactions with their respective cell surface receptors and that the effects of the activation of these receptors are mediated by different G-proteins. PMID- 2553000 TI - Receptor-ligand interaction: a new method for determining binding parameters without a priori assumptions on non-specific binding. AB - Analysis of receptor-ligand binding characteristics can be greatly hampered by the presence of non-specific binding, defined as low-affinity binding to non receptor domains which is not saturable within the range of ligand concentrations used. Conventional binding analyses, e.g. according to the methods described by Scatchard or Klotz, relate the amount of specific receptor-ligand binding to the concentration of free ligand, and therefore require assumptions on the amount of non-specific binding. In this paper a method is described for determining the parameters of specific receptor-ligand interaction which does not require any assumption or separate determination of the amount of non-specific binding. If the concentration of labelled free ligand is constant, a plot of Fu/(B0*-B*) versus Fu yields a linear relationship, in the case of a single receptor class, in which Fu is the concentration of unlabelled free ligand, B0* is the total amount of labelled bound ligand in the absence of unlabelled ligand and B* is the total amount of labelled bound ligand in the presence of an unlabelled ligand concentration Fu; all of these data are readily obtained from binding studies. This linear relationship holds irrespective of the amount of non-specific binding, and the values for receptor density, ligand dissociation constant and a constant for non-specific binding can be readily obtained from it. If the concentration of labelled free ligand is not a constant for all data points, data are first converted according to a straightforward normalization procedure to permit the use of this relationship. The presence of multiple receptor classes with dissociation constants in the range of the ligand concentrations used results in a negative deviation from this linearity, and therefore the presence of multiple receptor classes can be discriminated unequivocally from non-specific binding. Both theoretical and practical advantages of the present method are described. The method, which will be referred to as the linear subtraction method, is illustrated using the binding of tumour promoters and polypeptide growth factors to their specific cellular receptors. PMID- 2553001 TI - Interactions between lithium, inositol and mono-oleoylglycerol in the regulation of insulin secretion from isolated perifused rat islets. AB - In response to stimulation by 20 mM-glucose, 15 mM-4-methyl-2-oxopentanoate or 10 mM-glyceraldehyde, isolated perifused rat islets respond with brisk biphasic insulin-secretory responses. The inclusion of 10 mM-LiCl significantly decreased second-phase insulin release in response to all agonists. Inositol, at a concentration (10 mM) which has no effect on secretion in the presence of 2.75 mM glucose, restored significantly glucose-, 4-methyl-2-oxopentanoate- or glyceraldehyde-induced second-phase release from Li+-treated islets. The addition of the diacylglycerol kinase inhibitor mono-oleoylglycerol, at a concentration (25 microM) which has no stimulatory effect on insulin secretion in the presence of 2.75 mM-glucose, significantly amplified both the first- and second-phase insulin responses to 20 mM-glucose. This amplifying effect of mono-oleoylglycerol was readily reversible and dependent on Ca2+ influx into the beta-cell. Li+ decreased the amplified insulin response to 20 mM-glucose plus mono oleoylglycerol. Inositol restored release under this condition. These findings suggest that Li+ inhibits release by sequestering inositol into biosynthetically ineffective inositol phosphates. By limiting phosphoinositide resynthesis, the continued hydrolysis of phosphoinositides is diminished. Our results with mono oleoylglycerol suggest further that diacylglycerol content may play a critically important role in the regulation of both the first and second phases of insulin secretion. PMID- 2553002 TI - Effects of vanadate on protein kinases in rat hepatocytes. AB - In rat hepatocytes, vanadate modifies neither the intracellular concentration of cyclic AMP nor the --cyclic AMP/+cyclic AMP activity ratio for cyclic AMP dependent protein kinase. Vanadate can, however, counteract the increase in cyclic AMP and the increase in the --cyclic AMP/+cyclic AMP activity ratio of cyclic AMP-dependent protein kinase induced by glucagon. On the other hand, vanadate treatment of hepatocytes can produce a time- and concentration-dependent increase in cyclic AMP- and Ca2+-independent casein kinase activity. Maximal activation at the optimal time with 5 mM-vanadate was about 70% over control. A clear relationship was observed between the activation of casein kinase and the inactivation of glycogen synthase after vanadate treatment. These results suggest that casein kinase activity may be involved in vanadate actions in rat hepatocytes. PMID- 2553003 TI - Studies on the electron-transfer mechanism of the human neutrophil NADPH oxidase. AB - A superoxide-generating NADPH oxidase was solubilized from phorbol 12-myristate 13-acetate-activated human neutrophils with a mixture of sodium deoxycholate (0.125%, w/v) and Lubrol-PX (0.125%, v/v). The solubilized preparation contained FAD (577 pmol/mg of protein) and cytochrome b-245 (479 pmol/mg of protein) and produced 11.61 mol of O2-./s per mol of cytochrome b (340 nmol of O2-./min per mg of protein). On addition of NADPH, the cytochrome b-245 was reduced by 7.9% and the FAD by 38% in the aerobic steady state; NADH addition caused little steady state reduction of cytochrome b and FAD. In this preparation, and several others, the measured rate of O2-. production correlated with the turnover of cytochrome b calculated from the extent of cytochrome b-245 reduction under aerobic conditions. Addition of diphenyleneiodonium abolished the reduction of both the FAD and cytochrome b-245 components and inhibited O2-. production. The haem ligand imidazole inhibited O2-. generation and cytochrome b reduction while permitting FAD reduction. These results support the suggestion that the human neutrophil NADPH oxidase has the electron-transport sequence: NADPH----FAD--- cytochrome b-245----O2. PMID- 2553004 TI - Ionic-strength-dependence of the oxidation of native and pyridoxal 5'-phosphate modified cytochromes c by cytochrome c oxidase. AB - The ionic-strength-dependences of the rate constants (log k plotted versus square root of 1) for oxidation of native and pyridoxal 5'-phosphate-modified cytochromes c by three different preparations of cytochrome c oxidase have complex non-linear character, which may be explained on the basis of present knowledge of the structure of the oxidase and the monomer-dimer equilibrium of the enzyme. The wave-type curve (with a minimum and a maximum) for oxidation of native cytochrome c by purified cytochrome c oxidase depleted of phospholipids may reflect consecutively inhibition of oxidase monomers (initial descending part), competition between this inhibition and dimer formation, resulting in increased activity (second part with positive slope), and finally inhibition of oxidase dimers (last descending part of the curve). The dependence of oxidation of native cytochrome c by cytochrome c oxidase reconstituted into phospholipid vesicles is a curve with a maximum, without the initial descending part described above. This may reflect the lack of pure monomers in the vesicles, where equilibrium is shifted to dimers even at low ionic strength. Subunit-III-depleted cytochrome c oxidase does not exhibit the maximum seen with the other two enzyme preparations. This may mean that removal of subunit III hinders dimer formation. The charge interactions of each of the cytochromes c (native or modified) with the three cytochrome c oxidase preparations are similar, as judged by the similar slopes of the linear dependences at I values above the optimal one. This shows that subunit III and the phospholipid membrane do not seem to be involved in the specific charge interaction of cytochrome c oxidase with cytochrome c. PMID- 2553005 TI - Protein phosphatase composition in the smooth muscle of guinea-pig ileum studied with okadaic acid and inhibitor 2. AB - Using okadaic acid, a potent inhibitor of type 2A and type 1 protein phosphatases, and inhibitor 2, an intrinsic inhibitory factor of type 1 phosphatase, we characterized the phosphorylated myosin light-chain (PMLC) phosphatase activity in the smooth-muscle extracts of guinea-pig ileum. In the intact fibres the control activity was 254 +/- 13 nmol of Pi/min per g wet wt. (n = 15) against 32P-labelled PMLC (4 microM) from chicken gizzard. The following phosphatase fractions were identified: an inhibitor-2-sensitive (type 1) fraction (fractional activity = 35%), a Mg2+-dependent and okadaic acid-insensitive (type 2C) fraction (17%), and two type 2A-like fractions that had different susceptibility to okadaic acid. The type 2A-like fraction with lower affinity to okadaic acid accounted for 30% of the control activity. After the cell membrane was permeabilized by Triton X-100, more than 60% of this fraction remained and accounted for about 90% of the total activity, whereas the other fractions were nearly abolished. The type 2A-like fraction may be bound to some intracellular structure such as contractile proteins. PMID- 2553006 TI - Differential mechanisms of inositol phosphate generation at the receptors for bombesin and platelet-derived growth factor. AB - We investigated the mechanism(s) whereby activation of a growth-factor receptor typically endowed with tyrosine kinase activity, such as the platelet-derived growth factor (PDGF) receptor, triggers phosphoinositide hydrolysis. In Swiss 3T3 cells permeabilized with streptolysin O, an analogue of GTP, guanosine 5'-[gamma thio]triphosphate, was found to potentiate the coupling of the bombesin receptor to phospholipase C. In contrast, the activation of the enzyme by PDGF occurred in a GTP-independent manner. Moreover, the inactive analogue of GTP, guanosine 5' [beta-thio]diphosphate, significantly inhibited the bombesin-induced InsP3 generation, whereas it did not decrease the same effect when stimulated by PDGF. PMID- 2553007 TI - [The ectophosphatase activity of cultured endothelial cells of calf aorta and the effect of drugs on ecto-ATPase]. AB - Cultivated endothelial cells of calf aorta (line BKEz-7) possess an effective ectophosphatase system (enzyme activities: ATPase 38.0 +/- 10.2; ADPase 9.2 +/- 4.2; 5'-nucleotidase 4.1 +/- 2.6 fmol/cell.min). Drugs with central depressive activity such as promazine, chlorpromazine, and meprobamate inhibit the activity of the ecto-ATPase. A possible connection between the inhibitory activity on the ecto-ATPase and their central depressive effects is discussed. PMID- 2553008 TI - Renal effects of vanadate in rats. AB - In conscious rats, administration of vanadate is followed by a distinct and statistically significant diuretic effect. The increase in renal excretion of sodium dominates in comparison with other electrolytes. After i.p. administration vanadate is effective for 30 to 60 min. In anesthetized rats an excessive saline load and i.v. administration of vanadate do not prolong the renal effectiveness of vanadate. A long lasting diuretic effect is measurable during a constant infusion of vanadate. p-Aminohippurate transport in renal cortical slices can be inhibited by adding vanadate to the incubation medium. Vanadate administered in vivo in diuretically effective doses influences the p-aminohippurate transport in consectively prepared renal cortical slices. A 50% inhibition of Na+/K+-ATPase can be provoked by i.p. administration of 0.6 mg vanadate/100 g b.m. whereas the maximal increase in renal excretion of sodium is measurable following 0.5 mg vanadate/100 g b.m. PMID- 2553009 TI - Leukotriene A4 hydrolase activity of human airway epithelial cells. AB - Human tracheal epithelial cells were incubated with LTA4 and metabolic products were identified in extracted supernatants by high pressure liquid chromatography, ultraviolet spectroscopy, and gas chromatography-mass spectrometry. In the presence of epithelial cells, LTA4 was converted to LTB4, but not to LTC4 or LTD4. Maximum LTB4 was released at an LTA4 concentration of 3 microM and had occurred by 30 min. LTB4 release was increased in the presence of albumin, but was not affected by extracellular calcium or A23187. This LTA4 hydrolase activity had a slower time course and could not be clearly inactivated by repeated exposure to substrate as is the case for previously described LTA4 hydrolase enzymes. This hydrolase appears to have novel biochemical characteristics. PMID- 2553010 TI - Competitive peptide antagonists of ANF-induced cyclic guanosine monophosphate production. AB - Atrial natriuretic factor (isoleucine ANF 101-126), cleaved ANF (isoleucine ANF 101-105/106-126) and des (Gln 18, Ser 19, Gly 20, Leu 21, Gly 22) ANF 4-23-NH2 (C ANF 4-23) stimulated cyclic guanosine monophosphate production (cGMP) by rat aortic vascular smooth muscle cells (VSMC) in culture. Cleaved ANF and ANF C4-23 also antagonised or diminished the response to ANF 101-126. Agonist and antagonist actions of both peptides were dose-related. In contrast, prepro ANF (104-123), an ANF precursor fragment, exhibited no agonist or antagonist effect on cGMP production. PMID- 2553011 TI - Thrombin's effects on osteoblastic cells. I. Cytosolic calcium and phosphoinositides. AB - Thrombin, a blood coagulation factor, has been shown to be a very effective in vitro bone resorbing agent whose mechanism of action on osteoblastic cells remains to be elucidated. In the present study, the effects of highly purified human thrombin on Saos-2 and G292 cells, two human osteoblast-like osteosarcoma cell lines, were investigated. Thrombin (0.6-16 U/ml) caused a significant, dose dependent increase in osteoblastic cell proliferation. Thrombin also elicited a dose-dependent increase in cytosolic calcium concentration in both Saos-2 and G292 cells (maximal increases were 38% and 200% over baseline, respectively). Addition of thrombin to the osteoblast-like cells resulted in significant time- and dose-dependent changes in phosphoinositide levels: the percentage of inositol monophosphate levels were decreased, whereas the percentage of inositol bisphosphate, inositol trisphosphate and inositol tetrakisphosphate levels were increased. The relative magnitude of the changes in phosphoinositide levels was similar to the changes in cytosolic calcium concentration. These results suggest that thrombin's mechanism of action on bone cells may involve increases in cytosolic calcium levels and in phosphoinositide metabolism. PMID- 2553012 TI - Intracellular copper transport in cultured hepatoma cells. AB - The distribution of copper in lysates prepared anaerobically from copper resistant hepatoma cells radiolabeled with 67Cu was examined in pulse-chase experiments. Initially, the majority of the radioactivity (greater than 85%) coeluted with copper-metallothionein. As the chase time increased there was a gradual loss of 67Cu from metallothionein, with a concomitant increase in the level of 67Cu-labeled glutathione. There was also an increase in 67Cu incorporation into superoxide dismutase. These results suggest that the chelation of copper by metallothionein from a copper-glutathione complex (Freedman, J. H., Ciriolo, M. R., and Peisach, J. (1989) J. Biol. Chem. 264, 5598-5605) is a reversible process. Further, they demonstrate that the copper bound to metallothionein is not permanently sequestered, but can be incorporated into other copper proteins. PMID- 2553013 TI - Solubilization and identification of human placental endothelin receptor. AB - Endothelin-1 (ET-1) receptor was identified on the membranes from human placenta and 66% of original binding activity in the membranes was solubilized with 0.75% (w/v) CHAPS. Binding studies of the solubilized membranes using 125I-ET-1 indicated the presence of a single class of high-affinity binding sites with an apparent Kd of 760 pM and a Bmax of 1.8 pmol/mg of protein. The binding was inhibited by addition of unlabeled ET-1 and ET-3 in dose dependent manner. The Ki values of solubilized membranes were 84 pM for ET-1 and 250 pM for ET-3, whereas particulate membranes had weaker affinities (Ki = 410 pM for ET-1, 2500 pM for ET 3). Calcium channel blockers such as nicardipine, verapamil and diltiazem did not affect the binding of 125I-ET-1. Affinity labeling of the particulate and solubilized membranes with CHAPS revealed a specific binding protein with a Mr of 32,000. PMID- 2553014 TI - High level expression of p55, a thyroid hormone binding protein which is homologous to protein disulfide isomerase in a retroviral vector. AB - To develop an efficient system for a high level expression of a human cellular thyroid hormone binding protein (p55) in eukaryotic cells, a full-length p55 cDNA was inserted into a Harvey murine sarcoma virus-derived vector (pHTBr) and transfected into mouse NIH 3T3 cells. The expressed p55 has a molecular weight of 55,000 and is recognized by the human specific anti-p55 monoclonal antibody. Similar to the endogenous p55, the expressed p55 is localized on endoplasmic reticulum and nuclear envelope. Moreover, p55 was specifically labeled by N bromoacetyl-3,3',5-triiodo-L-thyronine. Thus, the expressed p55 is structurally indistinguishable from the endogeneous p55. pHTBr was packaged into a virus with the aide of an amphotropic virus. Infection by pHTBr-containing virus yielded a 2 11 fold higher expression than the endogeneous p55 in NIH3T3, rat GH3, human HepG2 cells and a mouse monoclonal antibody secreting hybridoma. PMID- 2553015 TI - Proteins associated with mRNA in cells infected with herpes simplex virus. AB - The structure of messenger ribonucleoprotein (mRNP) complexes in herpes simplex virus type 1 (HSV-1) infected cells was analyzed by examining the proteins that could be crosslinked to polyadenylated mRNAs by irradiation of intact cells with ultraviolet light. The profiles of crosslinked proteins were qualitatively similar for mRNPs from mock infected and infected cells. However, infection with wild type HSV-1 caused a decrease in the abundance of a major 52 kda protein and an increase in a 49 kda protein. These changes were observed at early times after infection. They occurred following infection with wild type HSV-1 under conditions that blocked viral gene expression, but not following infection with the virion host shutoff mutant vhs 1. PMID- 2553016 TI - Developmental changes in the activities of phospholipase c, 3-kinase, and 5 phosphatase in rat brain. AB - The specific activities of phospholipase C, 3-kinase, and 5-phosphatase were measured in brain homogenates from rats at different developmental stages. The activities of 3-kinase and 5-phosphatase increased by 14-fold and 2-fold, respectively, during development from fetus to adult, while PLC activity remained constant. These results suggest that the metabolism of inositol phosphates varies widely during development. In young brain stimulated by an agonist, it is predictable that Ins(1,4,5)P3 lasts longer and its average concentration is higher than in adult brain. The opposite is true for both the lifetime and concentration of Ins(1,3,4,5)P4. These developmental changes will invariably affect the property of Ca2+ oscillation and the effective time during which cells respond to the Ca2+-mobilizing agonists. PMID- 2553017 TI - Tissue- and cell type-specific expression of mRNAs for four types of inositol phospholipid-specific phospholipase C. AB - The mRNA levels for four types of inositol phospholipid-specific phospholipase C (PLC) in various tissues and cell cultures have been studied by Northern analysis using cDNA probes for PLC isozyme I, II, and III [Sue, P.-G., Ryu, S.H., Moon, K.H., Sue, H.W., and Rhee, S.G. (1988) Proc. Natl. Acad. Sci. USA 85, 5419-5423 and Cell 54, 161-169], and the recently identified isozyme IV. All four types are ubiquitously expressed in rat tissues, but the levels of the mRNAs vary among tissues and cell lines. PLC-I mRNA levels are extremely high in brain and rat C6 glioma cells with lower levels in other tissues tested. PLC-II and -III have a more widespread distribution, with relatively high levels in brain, lung, spleen, thymus, and testis in the case of PLC-II, and in skeletal muscle, spleen, and testis for PLC-III. PLC-II and -III mRNAs were also detected in all cell lines examined except human promyelocytic HL60 cells. PLC-IV mRNA levels are extraordinarily high in spleen and HL60 cells. These results indicate that rat C6 glioma cells, together with most rat tissues, contain all four PLC isozymes. Other cultured cell types examined also contain two or three PLC isozymes except for HL60 cells, which contain only PLC-IV. The concomitant expression of PLC isozymes in cultured cells suggests a diverse function for PLC isozymes in single cells. PMID- 2553018 TI - Human red blood cells as bioreactors for the release of 2',3'-dideoxycytidine, an inhibitor of HIV infectivity. AB - 2',3'-Dideoxycytidine (ddCyd) is one of the most potent antiviral nucleosides for killing the human immunodeficiency virus (HIV). ddCyd is currently used in the treatment of severe HIV infections but due to its rapid clearance it must be administered to patients every 4 h reaching concentrations that are toxic. We have synthesized 2',3'-dideoxycytidine-5'-phosphate (ddCMP) as a prodrug, encapsulated it in human erythrocytes and found that it is dephosphorylated by endogenous pyrimidine nucleotidases and subsequently released by the cells as ddCyd. Encapsulated ddCMP does not affect erythrocyte metabolism and was not deaminated by cytidine deaminase. The dephosphorylation reaction has an apparent Km of 6mM, an optimum pH of 6.8 and is not inhibited by ATP or 2,3 bisphosphoglycerate. The efflux of ddCyd from the erythrocyte is a linear function of ddCyd concentration and relatively insensitive to nucleoside transporter inhibitors suggesting that ddCyd permeates the erythrocyte membrane predominantly by nonfacilitated diffusion. Thus, ddCMP-loaded erythrocytes might be used as endogenous bioreactors for ddCyd delivery in the treatment of HIV infection. PMID- 2553019 TI - Suppression of insulin-stimulated glucose transport in L6 myocytes by calcitonin gene-related peptide. AB - The binding of calcitonin gene-related peptide (CGRP) to L6 myocytes, the coupling of this receptor to adenylyl cyclase and the resultant effects on insulin-stimulated 2-deoxyglucose uptake were examined. L6 cells express specific binding sites for CGRP. Binding of human [125I]CGRP was inhibited by rat CGRP with an IC50 of approximately 10(-9) M. Synthetic human calcitonin at concentrations up to 10(-6) M had no effect on the binding of CGRP, suggesting that L6 cells express CGRP receptors, rather than calcitonin receptors which are also capable of binding CGRP. The CGRP receptor appeared to be coupled to adenylyl cyclase. Concentrations of CGRP greater than 3 x 10(-9) M increased the cellular content of cAMP. At 3 x 10(-8) M, CGRP increased cAMP 500-fold. CGRP at 10(-10) M and above suppressed the stimulation of 2-deoxyglucose uptake by insulin. Acute incubation of L6 cells with insulin stimulated 2-deoxyglucose uptake 1.6-fold, which was inhibited up to 70% by CGRP. Our results demonstrate that the specific binding of CGRP to L6 cells causes large increase in the cellular content of cAMP - and inhibition of insulin-stimulated 2-deoxyglucose uptake, but the differences in the dose-response curves suggest that the suppression of insulin action by CGRP cannot be solely explained by the increase in cAMP. PMID- 2553020 TI - Novel peptide inhibitor (SPAI) of Na+, K+-ATPase from porcine intestine. AB - Three unique inhibitors (SPAI-1, -+2, and -3) were first purified from porcine duodenal extract based on the Na+, K+-ATPase inhibitory activity. These peptide inhibitors had four disulfide bridges in common. The sequencing results of their S-carboxymethyl derivatives, lysilendopeptidase fragments, and chymotryptic peptides disclosed their entire primary structures. Both SPAI-2 and -3 consisted of 61 amino acids, respectively, and had almost the same sequences except for two amino acid substitutions, while SPAI-1 was found to lack the N-terminal twelve amino acid sequence of SPAI-2. The kinetics study revealed that SPAIs inhibited Na+, K+-ATPase by the competitive mode against Na+ and were uncompetitive with K+. PMID- 2553021 TI - Expression of cellular retinoic acid binding protein II (chick-CRABP II) in the chick embryo. AB - We previously demonstrated the presence of cellular retinoic acid binding protein II, chick-CRABP II, in chick embryos. In the present study, we investigated the distribution of chick-CRABP II in 14-day chick embryos by means of immunoblot analysis. Chick-CRABP II was expressed in skin, muscle, bone with tendon of the embryos, but not expressed in the nervous system. In adult chick tissues, chick CRABP II was not detected on immunoblotting; Chick-CRABP II in adults amounts to less than 10 ng/mg soluble protein. These observations suggest that chick- CRABP II is an embryonic protein involved in the development of specific tissues, such as bone, muscle and skin. PMID- 2553022 TI - Characterization of renal glomerular endothelin receptors in the rat. AB - Specific receptors for Endothelin (Et) have been identified in the glomerulus of rat kidney. Human 125I-ET binds to a single population of high affinity receptors in glomerular membranes from normal rats with a mean equilibrium dissociation constant of 200 pM. The binding was time and temperature-dependent, saturable and reversible. The Et-receptor complex was not affected by either vasopressin, atrial natriuretic factor or angiotension II. PMID- 2553023 TI - In vitro phosphorylation of the tumor suppressor gene RB protein by mitosis specific histone H1 kinase. AB - The major components of the mitosis-specific histone H1 kinase are CDC2 kinase and cyclin and the consensus amino acid sequence for phosphorylation by this enzyme has been proposed. We have noted the presence of such sequences in six sites of the tumor suppressor gene RB protein and determined whether or not RB protein is in fact phosphorylated by this kinase. Highly purified enzyme was used for this purpose. HeLa cell extracts immunoprecipitated with anti-RB antiserum as well as RB proteins expressed in E. coli cells were shown to be phosphorylated by this kinase in vitro. Synthetic peptides for the six expected sites were also phosphorylated. These results suggest the possibility that the function of RB protein is regulated by CDC2 kinase. PMID- 2553024 TI - The 5'region of the rat phosphoenolpyruvate carboxykinase gene confers pH sensitivity to chimeric genes expressed in renal and liver cell lines capable of expressing PEPCK. AB - The 5' flanking regions of the rat phosphoenolpyruvate carboxykinase gene were used to form chimeric gene constructs with the human growth hormone gene. These constructs were transfected into several renal and one liver cell line and the production of growth hormone (HGH) measured by immunoassay. Cyclic-AMP and glucocorticoid responsiveness of HGH production was observed in all cell lines. In two lines, the rat NRK52E renal epithelial line and the rat H4IIE hepatoma cell line, both capable of expressing PEPCK, lowering extracellular pH increased HGH production several fold. Comparison of hormone and pH effect on cells transfected with a thymidine kinase promoter-HGH chimera indicated that the PEPCK 5' flanking region effects were specific. Thus, part of the pH responsiveness of the PEPCK gene in vivo may be attributed to properties of the 5' flanking regions. PMID- 2553025 TI - Suppression of human synovial cell proliferation by dihomo-gamma-linolenic acid. AB - Prostaglandin E1 (PGE1) and oils enriched in its precursor fatty acids suppress inflammation and joint tissue injury in several animal models. Since synovial cell proliferation is a hallmark of rheumatoid arthritis, we studied the effect of dihomo-gamma-linolenic acid (DGLA), an immediate precursor of PGE1, on the growth of human adherent synovial cells (ASC) in tissue culture. When stimulated by appropriate concentrations of recombinant interleukin-1 beta (rIL-1 beta), ASC proliferate and produce PGE. DGLA-enriched medium suppressed both baseline and rIL-1 beta-stimulated ASC growth fivefold, compared with medium supplemented with arachidonic acid. Indomethacin reduced the effect of the DGLA. Synovial cells incorporated the DGLA, and rIL-1 beta-stimulated cells that were incubated with DGLA exhibited a 14-fold increase in PGE1 (to 25.2 +/- 6.0 ng/ml, mean +/- SD) and a 70% decrease in PGE2 (to 25.2 +/- 4.2 ng/ml) compared with cells in control medium. At equivalent concentrations (5 x 10(-7) M), PGE1 increased the level of cellular cAMP to a greater extent than did PGE2 (16.8 +/- 2.0 pmoles versus 4.3 +/- 1.9 pmoles, mean +/- SEM). Exogenous PGE1 was also a more effective inhibitor of cell growth. Similarly, cAMP concentrations in cells exposed to DGLA for 6 hours were greater than concentrations in arachidonic acid-enriched cultures (17.8 +/- 3.3 pmoles versus 2.1 +/- 2.0 pmoles). These observations suggest that DGLA can restrain ASC growth, an effect which may be due to its capacity to increase PGE1 production and subsequent cellular cAMP concentration. PMID- 2553026 TI - Human cytomegalovirus nucleic acids in tissues from congenitally infected infants. AB - In situ nucleic acid hybridization methods were used to study autopsy tissues from 2 infants with clinical or pathologic evidence of congenital cytomegalovirus infections: a premature infant with multiple congenital malformations and inclusion bodies in several organs and an infant with culture-proved symptomatic congenital cytomegalovirus infection. Cytomegalovirus nucleic acids were detected in lung, kidney, adrenal, thyroid, skin, pancreas, and anterior pituitary tissues. In the second infant, hybridization studies of neural tissues failed to reveal cytomegalovirus nucleic acids despite neuropathologic abnormalities consisting of extensive necrotizing ependymitis and periventricular cavitation. PMID- 2553027 TI - Pyruvate carboxylase deficiency: acute exacerbation after ACTH treatment of infantile spasms. AB - Pyruvate carboxylase deficiency results in congenital lactic acidosis. We report the significant finding in a child with infantile spasms controlled with adrenocorticotrophin hormone (ACTH) but who then developed severe lactic acidosis; pyruvate carboxylase deficiency was subsequently diagnosed. Blood lactate, pyruvate, and alanine levels were elevated, as well as cerebrospinal fluid alanine. Plasma alanine concentration was doubled by ACTH therapy. Fibroblasts contained extremely low pyruvate carboxylase activity. The patient died at 12 weeks of age after recurrent episodes of profound acidosis. At autopsy, the brain manifested cystic degeneration and demyelination. Pyruvate carboxylase deficiency is associated with neonatal onset of acidosis, delayed development, seizures, hypotonia, recurrent profound acidosis, and early death. The dramatic rise in plasma alanine content coincident with ACTH therapy suggest that ACTH played a role in precipitating the catastrophic metabolic acidosis. PMID- 2553028 TI - Beagle puppy model of perinatal asphyxia: blockade of excitatory neurotransmitters. AB - The N-methyl-D-aspartate receptor antagonist MK801 has been reported to prevent neuronal change in models of ischemia in adult animal systems. We studied the hypothesis that blockade of the N-methyl-D-aspartate receptor would prevent the depression of cerebral high-energy phosphates found in perinatal asphyxia without producing alterations in cerebral blood flow, and thus prevent neuropathologic damage. Newborn beagle puppies were anesthetized, tracheotomized, ventilated, and randomized to asphyxial insult (I = discontinuation of ventilatory support for 5 min) or no insult (NI) and drug treatment with MK801 (10 mg/kg intravenously) or an equal volume of saline (S). Puppies received MK801 or saline 15 min prior to I/NI. In S/I pups during insult, blood flow increased to brainstem structures but decreased elsewhere. MK801 had no effect on cerebral blood flow in either control or insulted puppies. 1H NMR studies demonstrated no effect of the MK801 on NI brains. Phosphocreatine levels were 1.7 +/- 0.1, 0.6 +/- 0.1, and 0.9 +/- 0.1 mmole/kg (mean: +/- S.D.) for the S/NI, S/I, and MK801/I pups, respectively. Cerebral lactate was 1.3 +/- 0.2, 3.0 +/- 0.7, and 2.0 +/- 0.4, respectively. The pH fell 0.8 units in the S/I puppies, compared to 0.4 units in the MK801/I puppies. We conclude that pretreatment with the N-methyl-D-aspartate receptor antagonist MK801 in part protects the developing brain against severe metabolic insult. PMID- 2553029 TI - Intracranial tumors and West syndrome. AB - Two infants, 6 and 7 months of age, are reported with both West syndrome and cerebral tumors. In both patients, initial neurologic examinations were normal and tumor diagnoses were determined through routine imaging studies. Initial response to adrenocorticotrophic hormone treatment did not differ from that observed in patients with cryptogenic West syndrome. Early treatment of West syndrome and resection of tumor, when possible, may offer successful treatment of patients with both benign brain tumors and West syndrome. PMID- 2553030 TI - [Viral origin of postnatally acquired inner ear damage]. AB - Experimental and clinical data exist which point at a possible role of herpes virus infections in postnatally acquired inner ear disturbances. The presence of viral DNA in the inner ear and vestibular organ of experimentally infected mice and pigs suggests that not only primary infections but also the reactivation of latent herpes viruses can cause severe inner ear damage. Serological examinations are presented and therapeutic considerations discussed. PMID- 2553031 TI - [Cytomegalovirus infection and inner ear deafness]. AB - Bilateral sensorineural hearing loss of patients is correlated with elevated antibody titres against cytomegaly virus in serum in children as well as in adults. Unilateral deafness correlates with mumps infection. This is shown in a German population as well as in a Mediterranean group of patients. There is no evidence between antibody titer of Coxsackie virus, adenovirus and herpes virus or toxoplasma gondii interaction and sensorineural hearing loss neither unilaterally nor bilaterally. PMID- 2553032 TI - Non-adrenergic non-cholinergic nervous control of airways. AB - Traditionally, the tone of bronchial smooth muscle is mediated through the balance of two autonomic nervous systems. The existence of a third non-adrenergic non-cholinergic (NANC) nervous system in the lung has been demonstrated in animals. In contrast to high density parasympathetic innervation of smooth muscle, no evidence of adrenergic nerves was found in human airways. Electrical field stimulation of tracheal strips in guinea-pig produced an initial contraction followed by a relaxation. The contraction was blocked by atropine but the relaxation was not inhibited by a beta-adrenergic blocking agent. Immunohistochemical studies have shown that vasoactive intestinal peptide (VIP) like immunoreactive nerves are present in the central nervous system and in the peripheral neuronal system of mammals including the human respiratory tract. Autoradiographic and immunocytochemical studies have demonstrated a high density of VIP receptors in airway epithelium, submucosal glands, pulmonary vessels and smooth muscle, especially in large airways. Functional studies in humans have confirmed the regulating role of VIP principally in the large airway. PMID- 2553033 TI - Nedocromil sodium (Tilade): a review of preclinical studies. AB - Nedocromil sodium (Tilade) is a novel pyranoquinoline dicarboxylic acid, developed for the treatment of reversible obstructive airway disease (ROAD). The drug exhibits specific anti-inflammatory properties when administered topically to the airways. These properties are illustrated by the activity of the drug in a number of in vitro and in vivo models. Thus, in vitro, nedocromil sodium inhibits activation of, and mediator release from, a wide range of inflammatory cells (neutrophils, eosinophils, macrophages/monocytes, mast cells and platelets) in both animals and man, initiated by either specific or non-specific stimuli. In vivo, the drug is effective in models of anaphylactic bronchospasm, increased vascular permeability, cellular influx, late reaction and bronchial hyperreactivity. Although as yet the mechanism of action has not been fully clarified, nedocromil sodium (Tilade) has demonstrated unequivocal efficacy in the treatment of ROAD in controlled therapeutic studies of up to twelve weeks in duration. PMID- 2553034 TI - Nedocromil sodium: a review of clinical studies. AB - Nedocromil sodium is a new compound for the long-term treatment of bronchial asthma. Its effect has been studied in prevention of bronchospasm induced by a variety of stimuli showing a broader spectrum of activity than SCG. Its clinical efficacy has been demonstrated in a number of trials involving patients regardless of asthma type. PMID- 2553035 TI - Reproducibility and comparison of quantitative DNA histogram features obtained with a scanning microdensitometer and a flow cytometer in breast cancers. AB - A pilot study compared DNA histogram features of ten invasive ductal breast cancers measured by three different techniques: in cytologic and histologic samples using a scanning densitometer and in suspensions of single nuclei using a flow cytometer. The reproducibility of measurements pertaining to quantitative ploidy factors was better for flow cytometry (FCM) than for static cytometry and was worst of all using histologic samples. The use of external reference cells for the precise determination of the diploid value in measurements of paraffin embedded single nuclei proved to be pointless. Of all static histogram descriptors, the modal value was the most reproducible; however, this feature could not distinguish between diploid and near-diploid tumors. In contrast, using the descriptors of (1) diploid peak width, (2) coefficient of variation of the integrated optical density values and (3) percentage of cells exceeding 2.5c from histograms derived from static measurements, it was possible to distinguish between tumors that were characterized as diploid and near-diploid by FCM (chi square P = .01). This is the more important because subjective analysis of the static histograms of these tumors failed to identify a cell population deviating from the diploid value in spite of the fact that the FCM DNA index (DI) varied from 1.12 to 1.23. Thus, quantitative histogram descriptors, as evaluated in this study, may add objective information to the measurement of DNA ploidy when using static DNA cytometry, especially in the case of near-diploid tumors. Duplicate FCM assessments of the DI showed the highest correlation of all features investigated (r = .99). PMID- 2553036 TI - Study of inhibition of platelet aggregation of 1,8-naphthyridine derivatives. AB - The antiplatelet activity of some 1,8-naphthyridine derivatives was studied. The compounds displayed different abilities in blocking the effects of ADP, collagen and arachidonic acid. Four of them showed an in vitro remarkable activity similar to that of dipyridamole and papaverine. PMID- 2553037 TI - Dipyrazolo[5,4-b:3',4'-d]pyridines. Synthesis, inhibition of benzodiazepine receptor binding and structure-activity relationships. AB - Some 2,6,8-trisubstituted dipyrazolo[5,4-b:3',4'-d]pyridin-3-ones related to the CGS series were synthesized and tested for their ability to displace [3H]flunitrazepam binding from bovine brain membranes. However the affinity for the benzodiazepine receptor of the tested compounds was lower than that of the CGS series, although it was comparable to that of chlordiazepoxide. It follows that some structure-activity relationships on the tested compounds can be drawn. PMID- 2553038 TI - 1,2,3-triazolo[4,5-f]quinolines. II. Preparation and antimicrobial evaluation of 6-ethyl-6,9-dihydro-1(2)(3)-R-1(2) (3)H-triazolo [4,5-f]quinolin-9-one-8 carboxylic acids as anti-infectives of the urinary tract (1). AB - Some 6-ethyl-1(2)(3)-R-1(2)(3)H-triazolo[4,5-f]quinolin-9-one-8-carboxy lic acids were prepared as novel analogues of oxolinic acid in order to evaluate the effect on antibacterial activity of the isosteric replacement of the dioxolic moiety with the triazole ring substituted in position 1 or 2. In vitro tests showed a good and selective activity against Escherichia coli (MIC 12.5 micrograms/ml) of compound (XVI). PMID- 2553039 TI - Recovery of solid-associated virus: evaluation of different procedures. AB - Three different extraction-concentration methods for virological analysis of sludge, marine and river sediments are studied. Enterovirus present in the concentrates were enumerated using the plaque technique with BGM cells. The results show that the indirect extraction technique isolated virus from the largest number of marine sediment samples. The direct extraction method is the best to recover virus from river sediments. Among the sludge samples, the number of positive samples did not vary much with the different techniques. It appears that there is no one universal method for virus concentration applicable to all categories of sediments and sludge. PMID- 2553040 TI - Suppressive effects of the bacterial immunostimulant OK-432 on the incidence of spontaneous thymic lymphoma in AKR mice. AB - The mean survival age of female AKR/J mice was significantly prolonged, the enlargement of thymus was markedly suppressed, and the proliferation of ecotropic and recombinant murine leukemia viruses was inhibited when 2-month-old female AKR/J mice were injected intraperitoneally with attenuated Streptococcus pyogenes, strain Su (OK-432) twice weekly for 8 weeks. However, these effects of OK-432 in 2-month-old female AKR/J mice were not seen in 5-month-old female AKR/J mice. The difference in the effectiveness of OK-432 in these animals probably depends on the difference in the degree of the proliferation of ecotropic and recombinant murine leukemia viruses in the thymus which consequently lead to thymic lymphoma. PMID- 2553041 TI - Nosocomial and endemic infections with a genome type of adenovirus type 5. AB - A unique genome type of adenovirus 5 was isolated from 37 specimens of 25 patients. DNA restriction analysis was performed with seven endonucleases. Part of the patients were treated at a bone marrow transplantation unit in Munich. Nosocomial spread, rather than activation of endogenous adenovirus infection, was likely for many of them. In addition, sporadic cases suggested an endemicity of this genome type in Bavaria. PMID- 2553042 TI - Opportunistic infections with coronavirus-like particles in patients infected with the human immunodeficiency virus? AB - From 35 patients infected with Human Immunodeficiency Virus (HIV) and belonging to various risk groups, 60 stool specimens were examined for the presence of Coronavirus-like particles (CVLP) using electron microscopy. CVLP were detected in 5 (8%) stool samples from 5 different patients (14%). Only one of the patients had diarrhoea. The five patients with CVLP-positive stools were all at advanced stages of HIV infection. The remarkable discrepancy between our data and another study, reporting a rate of 50% CVLP-positive HIV patients, most of them persistently shedding the virus, is discussed. PMID- 2553043 TI - Posttetanic potentiation in decentralized and nondecentralized superior cervical ganglia of the cat. AB - The present investigation was aimed to answer the following elementary, though important question concerning the sympathetic ganglion: Do the decentralized preganglionic terminals retain their full capacity to develop posttetanic potentiation (PTP) before substantial Wallerian degeneration takes place? Experiments were performed on the cat superior cervical ganglion in situ, and they followed a factorial design. The factors were: tetanization (supramaximal pulses, 0.2 ms, 24 Hz, 30 s), acute decentralization, and moderate hexamethonium blockade (5 mg/kg). Two levels were dealt with, namely, the indicated maneuvers were either performed or not performed. PTP was was measured in the S2 wave and the following variables were studied: decay constant, area under the curve and delay to summit occurrence. The analysis of variance showed that decentralization did not affect the development of PTP. Therefore, the nondegenerated terminals are fully capable of sustaining PTP, without the aid of the preganglionic cell bodies. PMID- 2553044 TI - Effects of chronic antidepressant and benzodiazepine treatment on corticotropin releasing-factor receptors in rat brain and pituitary. AB - We examined the effects of chronic treatment with antidepressants (imipramine or desipramine) or benzodiazepines (diazepam, alprazolam, or adinazolam) on modulation of corticotropin-releasing-factor (CRF) receptors in discrete areas of rat brain and in anterior pituitary. As previously reported, we found that chronic antidepressant treatment downregulated 5-HT2 serotonin and beta adrenergic receptors in cerebral cortex. Although there was a trend toward increased CRF binding in brain stem, striatum, cerebellum, hypothalamus, and frontal cerebral cortex following antidepressant treatment, the changes were only statistically significant in brain stem in imipramine-treated rats. In addition, no significant changes were seen in CRF binding in other brain regions including parietal/temporal cerebral cortex, olfactory bulb, hippocampus, and anterior pituitary. Following chronic benzodiazepine treatment CRF receptor binding was significantly decreased in the frontal cerebral cortex and hippocampus; although there was a trend for CRF receptors to be decreased in other brain areas and increased in anterior pituitary, the changes were not statistically significant. PMID- 2553045 TI - Cytomegalovirus infection after organ transplantation: an update with special emphasis on renal transplantation. AB - Cytomegalovirus infections are still the most important infectious complications after organ transplantation. Besides historical notes this review will deal with new aspects concerning the epidemiology of the CMV, diagnostic modalities of CMV infection, the delicate counterbalance between the immune system and the CMV, as well as the symptomatology of this infection. Furthermore, aspects like prophylaxis and new, promising therapeutic regimes for treatment of infection will be dealt with. Although this update is applicable for all types of solid organ transplantation, emphasis will be on renal transplantation. PMID- 2553047 TI - Epidermal growth factor: the receptor and its function. AB - Epidermal growth factor (EGF) is a small polypeptide hormone with mitogenic properties in vivo and in vitro. EGF elicits biologic responses by binding to a cell surface receptor which is a transmembrane glycoprotein containing a cytoplasmic protein tyrosine kinase. EGF responses are mediated by ligand binding and activation of this intrinsic protein kinase. The receptor can be phosphorylated by other protein kinases, and this may regulate receptor function. Stimulation of the receptor tyrosine kinase activity by ligand binding must regulate the activity of an as yet undefined molecule(s) responsible for transmitting a mitogenic signal to the nucleus. PMID- 2553046 TI - Leukocyte cell surface enzymology: CD45 (LCA, T200) is a protein tyrosine phosphatase. AB - During 1987, striking advances were made in defining the receptors and ligands for cell-to-cell adhesion interactions involving leukocytes. In 1988, two major leukocyte differentiation antigens, CD10 (cALLA) and CD45 (LCA, T200), were shown to be enzymes while two other markers, CD4 and CD8, were found to be associated with an enzyme. In this article, Ed Clark and Jeff Ledbetter discuss recent findings in the emerging area of leukocyte cell surface enzymology with emphasis on CD45, a membrane-associated protein tyrosine phosphatase (PTPase)2,3. PMID- 2553048 TI - Purification and characterization of a Mn2+/phospholipid-dependent protein phosphatase from pig brain membranes. AB - A Mn2+/phospholipid-dependent protein phosphatase has been identified and characterized from brain membranes. The phosphatase contains three subunits with molecular weights of 64,000, 54,000, and 35,000 in a 1:1:1 molar ratio. On gel filtration, the enzyme has an apparent molecular weight of approximately 180,000. The phosphatase was active on many substrates, including p-nitrophenyl phosphate, phosphotyrosine, phosphothreonine, phosphorylase a, myelin basic protein, histones, type 1 phosphatase inhibitor-2, microtubule tau protein, and synapsin I. To dephosphorylate phosphoproteins, the phosphatase was dependent on such acidic phospholipids as phosphatidylinositol and phosphatidylserine but not on neutral phospholipids such as phosphatidylcholine and phosphatidylethanolamine. The phospholipid-mediated activation of the phosphatase was time and dose dependent and could be reversed by Triton X-100 or gel filtration. Kinetic study further indicates that phospholipid was able to increase the Vmax of the phosphatase but had no effect on the Km value for substrates, suggesting a direct interaction of phospholipids with the phosphatase. Conversely, in order to dephosphorylate phosphoamino acids such as phosphotyrosine and phosphothreonine, this phosphatase was entirely dependent on Mn2+. Phospholipids had no effect on the dephosphorylation of phosphoamino acids, whereas Mn2+ had no effect on the dephosphorylation of phosphoproteins. It is concluded that this Mn2+/phospholipid dependent membrane phosphatase has two distinct activation mechanisms. The enzyme requires Mn2+ to dephosphorylate micromolecules, whereas acidic phospholipids are needed to dephosphorylate macromolecules. This suggests that Mn2+ and phospholipids may play a role in regulating the substrate specificity of this multisubstrate membrane phosphatase. PMID- 2553049 TI - Studies on adenosine triphosphate transphosphorylases. XVIII. Synthesis and preparation of peptides and peptide fragments of rabbit muscle ATP-AMP transphosphorylase (adenylate kinase) and their nucleotide-binding properties. AB - Two peptide fragments, derived from the head and tail of rabbit muscle myokinase, were found to possess remarkable and specific ligand-binding properties (Hamada et al., 1979). By initiating systematic syntheses and measurements of equilibrium substrate-binding properties of these two sets of peptides, or portions thereof, which encompass the binding sites for (a) the magnesium complexes of the nucleotide substrates (MgATP2- and MgADP-) and (b) the uncomplexed nucleotide substrates (ADP3- and AMP2-) of rabbit muscle myokinase, some of the requirements for binding of the substrates to ATP-AMP transphosphorylase are being deduced and chemically outlined. One requirement for tight nucleotide binding appears to be a minimum peptide length of 15-25 residues. In addition, Lys-172 and/or Lys-194 may be involved in the binding of epsilon AMP. The syntheses are described as a set of peptides corresponding to residues 31-45, 20-45, 5-45, and 1-45, and a set of peptides corresponding to residues 178-192, 178-194, and 172-194 of rabbit muscle adenylate kinase. The ligand-binding properties of the first set of synthetic peptides to the fluorescent ligands: epsilon MgATP/epsilon ATP and epsilon MgADP/epsilon ADP are quantitatively presented in terms of their intrinsic dissociation constants (K'd) and values of N (maximal number of moles bound per mole of peptide); and compared with the peptide fragment MT-I (1-44) obtained from rabbit muscle myokinase (Kuby et al., 1984) and with the native enzyme (Hamada et al., 1979). In addition, the values of N and K'd are given for the second set of synthetic peptides to the fluorescent ligands epsilon AMP and epsilon ADP as well as for the peptide fragments MT-XII(172-194) and CB-VI(126 194) (Kuby et al., 1984) and, in turn, compared with the native enzyme. A few miscellaneous dissociation constants which had been derived kinetically are also given for comparison (e.g., the Ki for epsilon AMP and the value of KMg epsilon ATP obtained for the native enzyme) (Hamada and Kuby, 1978), and the K'd measured for Cr3+ ATP [corrected] and the synthetic peptide I1-45 (Fry et al., 1985b). PMID- 2553050 TI - A human endonuclease incises ultraviolet-irradiated DNA at cytosines and oxidized DNA at thymines. AB - Both ultraviolet irradiation and oxidation of DNA produce a variety of pyrimidine base damages. A human endonuclease recognizes such altered bases on these DNA substrates. This human endonuclease incises ultraviolet-irradiated DNA exclusively at sites of photochemically modified cytosines. The precise sites of incision by the human enzyme were determined by DNA sequencing. Chemically oxidized DNA was incised exclusively at thymine loci. The degree of enzymic cleavage at cytosine photoproducts was identical at each site. However, the extent of incision at selected oxidized thymine residues varied within the DNA sequence. These results indicate that the distribution of thymine oxidative modifications is influenced by the neighboring DNA bases. PMID- 2553051 TI - The distribution of beta-adrenergic receptors in guinea pig lungs and their changes in experimental allergic asthma. AB - In this paper, the distribution of pulmonary beta-adrenergic receptors (beta receptors) in normal and experimental allergic asthmatic guinea pigs was determined using autoradiographic method. The results showed that the distribution of beta-receptors in the lung sections was widespread. There was a significant decrease of beta-receptor density in several tissue structures of asthmatic guinea pig lungs compared with the controls, a 23.73% decrease in beta receptor binding sites in bronchiolar smooth muscles and a 18.65% decrease in bronchial smooth muscles; a 23.53%, a 14.23% and a 17.16% decrease in bronchiolar epithelium, in bronchial epithelium and in alveolar epithelium respectively. The results indicated that the beta-receptor decrease in smooth muscles in experimental asthmatic guinea pig lungs might be one of the important factors which made the tension of smooth muscles increase. The relationship between the beta-receptor decrease of other sites and bronchial asthma needs to be studied further. PMID- 2553052 TI - Effects of sodium salicylate on the febrile response and increased levels of cyclic AMP in cerebrospinal fluid during endogenous pyrogen-induced fever in rabbits. AB - To further evaluate the causality between endogenous pyrogen (EP)-induced fever and cyclic adenosine-3',5'-monophosphate (cyclic AMP) level, the effects of sodium salicylate (SS) on the febrile response and increased levels of cyclic AMP in both cerebrospinal fluid (c.s.f.) and plasma during EP-induced fever in rabbits were observed. The results suggest that cyclic AMP is probably involved in the central mediation of EP-induced fever and that increased concentration of cyclic AMP in c.s.f. associated with EP-induced fever is not the result of temperature elevation but appears to be caused by the increased synthesis in the central nervous system. In addition it is confirmed that blood is impossibly a contributory source of increased cyclic AMP in c.s.f. during EP fever, and that SS may act subsequent to the increase in cyclic AMP. PMID- 2553053 TI - Characteristics of the primary reaction of hypocrellin A photosensitization. AB - The primary reaction of photosensitization of Hypocrellin A (HA) has been studied by using techniques of ESR, spin-trapping and spin-counteraction. The experiments show that HA is able to generate not only 1O2, but also O-2, .OH and HA- which are observed for the first time. The conversion of generating active oxygen into generating nonoxygen free radical is confirmed as well. Based on the characteristics of the primary reaction which generates these active substances (transient products), it is proposed that the photosensitized damage for the biological system by HA is probably related to not only 1O2 itself, but also a multiple effect from 1O2 as well as O-2, .OH and HA- free radical. PMID- 2553054 TI - [Therapeutic study of the effect of selective decontamination on microbial overgrowth syndrome of the small intestine]. AB - The microbial overgrowth syndrome of the small bowel (MOS) is characterized by clinically found symptoms of increased metabolic activities of microorganisms existing in a great number in the intestinal juice of these patients. We treated 15 patients who suffered from MOS following a modified scheme of selective intestinal decontamination (SID) with oral doses of BerlocombinR (trimethoprim/sulfamerazin) and polymyxin for three weeks. Comparative investigations of duodenal juices before and after treatment showed unchanged high germ-counts (total aerobes and total anaerobes up to 10(7) germs/ml juice). Only the increase of Clostridium (up to 10(6)/ml) was statistically significant which has to be disapproved. In most of the patients there was no subjective improvement. SID in the examined manner is not able to cure a microbial overgrowth in the upper gastrointestinal tract or to reduce facultatively pathogenic microorganisms. PMID- 2553055 TI - [Seroconversion with trivalent Sabin vaccine in children less than four years of age]. AB - A group of 58 children under 4 years old, were vaccinated with trivalent oral poliovirus vaccine Sabin (OPV), in two sanitary areas of Chiapas, Mexico. The complement of procedures for the maintenance of the cold chain, and the changes in the potency of the vaccine prior and after the vaccination were evaluated. Nurses got a blood sample of 3cc and other one, four weeks after vaccination in order to evaluate the seroconversion rate attributable to Sabin vaccine, which was analyzed through neutralization serologic test. Moreover the investigation team got a sample of feces one day prior to vaccination and other one every week in the following three weeks to vaccination date in order to isolate the poliovirus in the feces of the children. The high pre-vaccination seropositivity suggests that in these areas exist a wide circulation of poliovirus, which probably were vaccine viruses and in less proportion wild viruses. The seroconversion rates were low for the three types of poliovirus, nevertheless 83% of the children had at least one of the three possibilities of seroconversion: unique, double or triple. Children with pre-vaccine seropositive probably had a secondary immune response. PMID- 2553056 TI - Deficits in conditioned heart rate and taste aversion in area postrema-lesioned rats. AB - Previous studies have shown that area postrema (AP) lesions cause deficits in conditioned taste aversion in the rat. They also lead to chronically lowered heart rate which can be reversed by the animals' increased appetite for and ingestion of hypertonic saline. Although not previously examined in conditioned taste aversion, changes in autonomic nervous system activity as reflected in heart rate may be an important aspect of conditioning. The present study investigated the effects of AP lesions on heart rate conditioned responses (CRs) and unconditioned responses (UCRs). Two groups of AP lesioned and sham-operated rats, one that did and one that did not drink saline solution to raise heart rate, were studied. Both LiCl and scopolamine, which have opposite effects on heart rate, were the unconditioned stimulus (UCS) agents in two separate studies. In intact rats, LiCl-mediated conditioned taste aversion was associated with decreased conditioned stimulus (CS) intake and decreased heart rate Both effects were blunted by AP lesions, although all rats displayed heart rate UCRs to LiCl. The AP rats that drank saline behaved like intact rats exhibiting both a conditioned taste aversion and conditioned heart rate responses to the CS. Although CS intake decreased, no heart rate CRs developed with scopolamine. Scopolamine-mediated conditioned taste aversion was attenuated in both saline and non-saline drinking AP-lesioned groups. Thus, when conditioned taste aversion was associated with heart rate CRs, the AP lesion-induced deficit was counteracted by saline ingestion. Conversely, when there were no heart rate CRs, conditioned taste aversion was disrupted by the lesion regardless of saline ingestion. PMID- 2553057 TI - Effects of dose, interdose interval, and drug-signal parameters on morphine analgesic tolerance: implications for current theories of tolerance. AB - Some unique predictions of a dual-process priming model of morphine analgesic tolerance were tested. Two experiments in which morphine injections during a tolerance acquisition phase were accompanied by nociceptive testing on a hot plate, confirmed the predictions that tolerance acquisition, retention, and environment-specificity would be augmented under signaled drug conditions when low doses or long interdose intervals (IDIs) were used but not when high doses or short IDIs were used. The implications for current alternative theories of morphine tolerance are discussed. PMID- 2553058 TI - Tolerance to the antinociceptive effect of morphine in spinally transected rats. AB - The effect of a spinal transection on morphine-induced tolerance in rats was examined with the tail withdrawal reflex (tail flick; TF), elicited by noxious thermal stimulation. Intact rats became tolerant to sc morphine injections (3.0 mg/kg) if they were tested on the TF after each injection. Morphine administration alone did not produce tolerance; tail flick tests alone did, though not always to a significant extent. However, when morphine only, TF tests only, or both were administered prior to transection (acute spinal rats), all groups were tolerant when tested 1 day after spinalization. When the same treatments were administered to rats 3 weeks after spinal transection (chronic spinal rats), neither morphine nor TF tests alone produced tolerance. Chronic spinal rats became tolerant only if they were tested after each injection. These results suggest, first that tolerance develops at the spinal cord as a result of either chronic opiate exposure or performance of the nociceptive response but that, intact rats, tolerance is inhibited or suppressed by a supraspinal action of morphine. Second, the fact that chronic spinal rats did not become tolerant to morphine or TF tests alone suggests either that such tolerance is mediated by descending input or that spinal transection produces intrinsic changes in the spinal cord that preclude the development of tolerance induced only by opiate or behavioral stimulation. PMID- 2553059 TI - Ontogeny of noradrenergic effects on ultrasonic vocalizations in rat pups. AB - The ontogeny of noradrenergic effects and the interaction of opioid and noradrenergic systems on vocalizations in rat pups from Day 10 to Day 18 were evaluated. Day 10 pups given clonidine (0.05 or 0.5 mg/kg) ip showed a sustained high level of calling throughout a 25-min isolation period that was reversed with yohimbine (0.1 mg/kg). Day 15 pups showed identical profiles with a lower baseline rate. Day 17 pups' calls were differentially affected according to dose; Day 18 pups reduced vocalizing with clonidine. In addition, it was found that at all ages when clonidine increased calling during isolation, the pups vocalized in the nest as well. Naltrexone, an opioid antagonist, lost its effectiveness to increase vocalizations after Day 15 unless it was given subsequent to clonidine. These results suggest that pups' vocalizations are differentially affected by noradrenergic and opioid stimulation or inhibition with developmental changes. PMID- 2553060 TI - Opioid action on response level to a danger stimulus in the crab (Chasmagnathus granulatus). AB - When a passing shadow is presented to the crab (Chasmagnathus granulatus), an escape response is elicited that habituates after repeated stimulation. Results of previous papers suggest that this habituation might be mediated by endogenous opiates, entailing the postulate that opiates may inhibit the response to a danger stimulus. This contention is tested herein. Two trials (T1, T2) of shadow stimulation were given 30 min apart, and the response activity was recorded. In Experiment 1 a range of morphine doses (0, 25, 50, 75, and 100 micrograms/g) were injected into crabs immediately after T1. Analysis of the M response values showed a morphine dose-dependent reduction in the crab's reactivity to the danger stimulus in T2. In Experiment 2, groups of crabs were injected with 1 of 4 solutions; saline, 70 micrograms/g morphine, 70 micrograms/g morphine plus 1.6 micrograms/g naloxone. Results suggest that morphine acts through an opiate receptor to inhibit the crab's response level to a danger stimulus. Three possible mechanisms of the morphine effect are discussed, and the most viable one suggests a central site of action. PMID- 2553061 TI - Cervical mucus enzymes as markers of the woman's fertile period. PMID- 2553062 TI - Progressive multifocal leukoencephalopathy. PMID- 2553063 TI - Vaccination. PMID- 2553064 TI - Immunodeficiency associated with bone marrow transplantation. PMID- 2553065 TI - Anti-receptor antibodies. PMID- 2553066 TI - Graft-versus-host disease and viral infections. PMID- 2553067 TI - The transmission of disease to allograft recipients via donor organs. PMID- 2553068 TI - Autoradiographic evidence for the localization of high affinity neurotensin binding sites on dopaminergic nerve terminals in the nigrostriatal and mesolimbic pathways in rat brain. AB - Monoiodo[125I]Tyr3-neurotensin binding in coronal and sagittal brain sections from rats that had received a unilateral 6-hydroxydopamine lesion of the median forebrain bundle, revealed a complete loss of neurotensin binding (NTH)-sites in the caudate nucleus of the lesioned side. Moreover, in the lesioned side, 83% of the NTH-sites disappeared also from the nucleus accumbens and 65% from the olfactory tubercle. NTH-sites subsisted partly in the Islands of Calleja and no significant alteration was observed in the rhinal sulcus and in the cingulate cortex. We conclude that, in the terminal area of the nigrostriatal pathway, all functional NTH-sites are located on presynaptic dopaminergic nerve endings. In the terminal areas of the mesolimbic pathway, the vast majority of NTH-sites are also located on dopaminergic nerve endings. Only a few NTH-sites may possibly be located on non-dopaminergic nerve endings in the olfactory tubercle. NTH-sites can therefore be considered as specific markers of the dopaminergic nerve terminals in both the nigrostriatal and mesolimbic pathways. PMID- 2553070 TI - Disturbances of transmembranous sodium transport systems induced by ethanol in human erythrocytes. An approach to the pressor effect of alcohol. AB - Several epidemiological and clinical studies have established a clear association between alcohol consumption and hypertension. The mechanism of the pressor effect of ethanol is not well understood. We studied the in vitro effects of increasing amounts of ethanol on different Na+ transport systems from human erythrocytes. Ethanol at concentrations higher than 32 mmol/L stimulated ouabain-sensitive Na+ efflux, the Na+ efflux depending on the Na+:Li+ countertransport and Na+ leak. At the same concentrations, ethanol inhibited bumetanide-sensitive Na+ efflux. We conclude that, with the exception of Na+-K+ pump, alcohol-induced alterations and those observed in erythrocytes from essential hypertensives may overlap. Therefore, alcohol consumption could potentiate those genetic abnormalities of Na+ transport and contribute to the pathogenesis of essential hypertension. PMID- 2553071 TI - Effect of prolonged infusion of ANF in normotensive and hypertensive monkeys. AB - It is now recognized that bolus and short-term infusions of atrial natriuretic factor (ANF) into different species lead to a slight and transient decrease of blood pressure, while prolonged infusions cause a significant blood pressure reduction in hypertensive but not in normotensive rats. The present study was designed to evaluate the effects of prolonged ANF infusions on blood pressure and humoral parameters in normotensive and hypertensive African green monkeys (Cercopithecus aethiops). Human-ANF infusions (100 ng/kg.hr) in conscious, normotensive vervets for a period of 48 hours evoked highly significant decreases of blood pressure (from 124/65 to 104/53 mm Hg), plasma renin activity, aldosterone, and hematocrit. This fall in blood pressure was not accompanied by an increase of plasma cGMP levels at the end of the infusion. Forty-eight hours after the infusion was terminated, the decrease in blood pressure was still significant (97/46 mm Hg), as was the drop in aldosterone. In hypertensive monkeys, systolic blood pressure declined from 175 +/- 8 to 130 +/- 8 mm Hg, while diastolic pressure fell from 117 +/- 10 to 88 +/- 4 mm Hg. These data demonstrate that the chronic infusion of ANF in both normotensive and hypertensive vervets has more profound effects than does acute bolus administration, effects that persist for a prolonged period of time after discontinuation of the infusion. PMID- 2553072 TI - Peripheral nerve lesions in traumatic brain-injured patients. PMID- 2553069 TI - Insulin and insulin-like growth factor receptors in the nervous system. AB - Insulin and the insulin-like growth factors (I and II) are homologous peptides essential to normal metabolism as well as growth. These peptide hormones are present in the brain, and, based on biosynthetic labeling studies as well as evidence for local gene expression, they are synthesized by nervous tissue as well as being taken up by the brain from the peripheral circulation. Furthermore, the presence of insulin and IGF receptors in the brain, on both neuronal and glial cells, also suggests a role for these peptides in the nervous system. Thus, these ligands affect brain electrical activity, either as neurotransmitters or as neuromodulators, altering the release and re-uptake of other neurotransmitters. The insulin and IGF-I and -II receptors found in the brain exhibit a lower molecular weight than corresponding receptors on peripheral tissues, primarily caused by alterations in glycosylation. Despite these alterations, both brain insulin and IGF-I receptors exhibit tyrosine kinase activity in cell-free systems, as do their peripheral counterparts. Brain insulin and IGF-I receptors are developmentally regulated, with the highest levels appearing in fetal or perinatal life. However, the altered glycosylation of brain receptors does not appear until late in fetal development. The receptors are widely distributed in the brain, but especially enriched in the circumventricular organs, choroid plexus, hypothalamus, cerebellum, and olfactory bulb. These studies on the insulin and IGF receptor in brain, add strong support to the suggestion that insulin and IGFs are important neuroactive substances, regulating growth, development, and metabolism in the brain. PMID- 2553073 TI - CGS-19755 and MK-801 selectively prevent rat striatal cholinergic and gabaergic neuronal degeneration induced by N-methyl-D-aspartate and ibotenate in vivo. AB - The in vivo efficacies and potencies of various excitatory amino acid agonists in inducing cholinergic neuronal degeneration were compared following unilateral injections into the rat striatum. Kainic acid (KA), alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionic acid (AMPA), ibotenic acid (IBO), and N-methyl-D aspartic acid (NMDA) all produced dose-related decreases in choline acetyltransferase (ChAT) activity. The relative order of potency was KA greater than AMPA greater than IBO greater than NMDA. Quisqualic acid (QUIS) was about as potent as NMDA, but the maximal decrease in ChAT activity was less (36%). N acetylaspartyl-L-glutamate (NAAG) did not significantly decrease ChAT activity when up to 1,000 nmoles was injected. Approximate equitoxic doses of agonists were then used to examine the ability of i.p. administered CGS-19755 and MK-801 to prevent in vivo excitatory amino acid-induced cholinergic and GABAergic neuronal degeneration. NMDA-induced decreases in ChAT and glutamic acid decarboxylase (GAD) activities were prevented by CGS-19755 (10-40 mg/kg) and MK 801 (1-10 mg/kg). CGS-19755 (40 mg/kg) and MK-801 (10 mg/kg) did not prevent loss of ChAT or GAD induced by KA or AMPA, but did prevent the degenerative effects of IBO. This study shows that CGS-19755 and MK-801, two NMDA receptor antagonists that act by different mechanisms, are completely selective following systemic administration. Moreover, the in vivo excitotoxic effects of IBO are mediated at NMDA receptor sites that are blocked by these compounds. PMID- 2553074 TI - Treatment of oral cavity vascular malformations using the neodymium:YAG laser. AB - Although rare, hemangiomas and lymphangiomas of the oral cavity are difficult management problems. Surgical resection is still commonly thought of by many when considering treatment. Several articles in 1986 reported good results using the neodymium:yttrium-aluminum-garnet laser for these lesions. Using this laser, we have treated six patients with hemangiomas or lymphangiomas of the oral cavity over the past 4 years. Although most patients have required several treatments, the response has been excellent overall. This report confirms the results of previous studies. PMID- 2553075 TI - Snake venom components and their cross-reactivity: a review. AB - Snake venoms are complex mixtures of organic and inorganic compounds, many of which display biological activity. It has been demonstrated that antisera raised against whole venom or a single purified venom protein from one species of snake will react with proteins in the venom of other species. This cross-reactivity between species may have applications in determining snake phylogeny, but recent studies on the variation of venom components within a species make these evolutionary conclusions questionable. PMID- 2553076 TI - Cytochrome c methylation. AB - In this review, protein methylation is outlined in general terms, highlighting the major amino acids that are methylated and some of the proteins in which they are found. The majority of the review examines the methylation of cytochrome c at Lys-77 of lower eukaryotes as a possible model for methylation studies. Early work involving the purification and characterization of the methyltransferase responsible for this methylation indicated cytochrome c was methylated posttranslationally, yet prior to import into the mitochondria. Methylation in vitro occurred only at the in vivo methylation site and only on cytochrome c. Later studies using in vitro translated apocytochrome c revealed that methylated, as compared with unmethylated, apocytochrome c was imported preferentially into yeast, but not rat liver, mitochondria. Efforts to discover the reasons for this preference have shown that methylation of apocytochrome c dramatically lowers its isoelectric point (against a predicted increase) and decrease its Stokes radius. A possible mechanism for these differences involving the disruption of hydrogen bonds is presented here with space-filling models. Finally, the in vivo significance of this modification is also discussed. PMID- 2553077 TI - [Study of a comparative statistical analytical model for nucleotide sequences in the genome of viruses of the Papovaviridae family]. AB - In the present study a model for comparative analysis of nucleotide sequences has been developed, in order to evaluate statistical features of nucleotide distribution in DNA strands without any genetic relationship. Every DNA strand has been considered as a finite Markov chain; a matrix, whose elements represent the number of couplings between a nucleotide and the following one in 5'-3' direction, has been used for every DNA strand, and the statistical relationship has been detected by using Kendall's test. The genomes of Polyomavirus (strain A2) and DPV have been analysed by the proposed model; a substantial likeness between the behaviour of nucleotide distribution on all four DNA strands analysed has been shown; the strongest likeness concerned the complementary strands of Polyomavirus as well as the homologous sense strands of both viruses. PMID- 2553078 TI - Relationship between progesterone receptor, axillary node status and productive fibrosis in ductal infiltrating carcinoma of the breast. AB - The presence of progesterone receptor (PR), productive fibrosis (PF), axillary nodal status and tumor size are important prognostic variables in breast cancer. In this study we have analyzed the relationship between these four parameters in 78 ductal infiltrating carcinomas of the breast. No relationship was found between PF and the presence of lymph node metastatic disease: however, in the tumors with positive nodes, a limited metastatic diffusion (1-3 lymph nodes) was significantly associated with PF (p less than 0.05). Our study failed to demonstrate any relation between estrogen receptor (ER) and both presence and extension of nodal involvement; in contrast, the tumors with limited metastatic diffusion (1-3 lymph nodes) had a significantly higher prevalence of PR positivity (p less than 0.05). Furthermore, PF was strongly associated with PR (p less than 0.001) and less well with ER (p less than 0.05). The simultaneous presence of PR and marked PF was strongly correlated with limited metastatic involvement of the axilla (p less than 0.007). Tumor size was correlated with the number of positive nodes (p less than 0.001), but not with PR or PF. The results demonstrate that PR status, PF and tumor size are related to limited metastatic diffusion of the axilla: furthermore PF and PR have been shown to be strongly related and we have demonstrated that, at least in ductal infiltrating carcinoma, the simultaneous presence of them identifies a subset of tumors with low metastatic capacity independently of tumor size. PMID- 2553079 TI - [Benign infantile mitochondrial myopathy due to reversible cytochrome c oxidase deficiency: histological and biochemical analysis]. AB - We reported a 3-week-old girl with profound generalized weakness, requiring assisted ventilation. There was no consanguinity or family history of any neuromuscular disorders. Serum levels of GOT, GPT and CK were slightly elevated. Although the clinical manifestations mimicked those of Werdning-Hoffmann disease, a lack of the respiratory muscle involvement and the presence of high serum lactate levels suggested an underlying metabolic disorder. During the observation over the months, she gradually improved clinically and assisted ventilation was discontinued at 19 months. Muscle biopsies were performed at 4 and 19 months of age. The first biopsy showed an excessive mitochondrial aggregation and numerous ragged-red fibers. The second biopsy revealed rare ragged-red fibers. However there were definite neurogenic changes including type grouping and angulated fibers. Cytochrome c oxidase (CCO) stain was positive in less than 5% of fibers in the first biopsy but in all fibers in the second biopsy. Biochemical analysis using muscle specimens showed a decreased CCO activity; 18.5% of the control level in the first biopsy and 53.3% in the second biopsy. Immunocytochemistry showed the presence of immunologically reactive enzyme protein in both biopsies. Clinical manifestations of our patient were almost identical to those of two cases reported by DiMauro, et al (1983). The enzyme defect in this case was reversible in contrast to that in the fatal infantile form of CCO deficiency. Histochemical and biochemical bases for these differences were discussed. PMID- 2553080 TI - [Adenoid cystic carcinoma presenting as cavernous sinus syndrome: a clinico pathological study of two cases]. AB - Adenoid cystic carcinoma (ACC), usually arising in the major and minor salivary glands, is well known to otolaryngologists, but is rarely encountered by neurologists or neurosurgeons. We report two ACC patients who presented initially with cavernous sinus syndrome and in whom CT did not demonstrate apparent abnormalities. Case 1 is a 48-year-old man who first developed right cavernous sinus syndrome. The patient came to our hospital four years later. At this time, the only abnormality found on CT was the clouded sphenoid sinus on the left, which was interpreted as sinusitis. Two years later, the repeat CT revealed the enhancing lesions in the area of bilateral cavernous sinuses with bony destruction. The nasolaryngological exploration of the left sphenoid sinus made the diagnosis of ACC. Case 2 is a 43-year-old man who developed unilateral cavernous sinus syndrome over three months. No radiological abnormality was apparent to the neurologist and neurosurgeons, however, the otolaryngologist detected a nasopharyngeal mass diagnosed as ACC. Histological examinations of the biopsy specimen from both cases revealed mixed cribriform, tubular, and solid patterns characteristic of ACC. Electron microscopic study in Case 1 demonstrated microvilli-containing epithelial tumor cells forming true glandular lumens. In these cystic spaces there were cellular debris and crystalloids consisting of hexagonally-arranged tubules with a diameter of 25 nm. Although thick basement membrane was found in the basal portion of the tumor cells facing the connective tissue, there was no so-called pseudocysts lined by replicated basal lamina. It is possible that the specimen examined electron microscopically is limited to a particular epithelial region.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553081 TI - [Two types of relaxation responses mediated by cyclic GMP in cerebral arteries]. AB - It has been reported that endothelium-derived relaxing factor (EDRF) possesses chemical and pharmacological properties that are indistinguishable from those of nitric oxide (NO). Moreover, NO is the active chemical species responsible for endothelium-independent vasodilation produced by nitrogen oxide-containing substances including glyceryl trinitrate (GTN). Both EDRF and GTN activate soluble guanylate cyclase and consequently increase cyclic GMP level in various artery preparations. However, there have been few reports regarding cyclic GMP accumulation induced by EDRF or GTN in canine cerebral arteries. Therefore, it was investigated whether EDRF and GTN cause vasodilation through the common pathway mediated by cyclic GMP in the canine basilar artery. The relaxation responses induced by EDRF or GTN were studied in the canine basilar artery by an isometric tension-recording method. EDRF was induced by calcium ionophore A 23187. A 23187 did not relax the vascular tissue in the absence of the endothelial cells. On the other hand, GTN did induce relaxation in either the presence or absence of endothelial cells. FeSO4 at 3 X 10(-5) M reversed A23187 induced relaxation, but not GTN-induced relaxation (N = 10). Since Fe2+ is able to catalyse the formation of O2- in oxygenated phosphate buffer, these findings suggest that Fe2+ antagonizes EDRF by inactivating it via the generation of O2-. By the addition of 10(-5) M methylene blue, both A 23187- and GTN-induced relaxations were reversed (N = 8). Moreover, pretreatment with 10(-5) M methylene blue augmented contractile responses to 3 X 10(-6) M prostaglandin F2 alpha (N = 5).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553082 TI - Establishment of an osteoinductive murine osteosarcoma clonal cell line showing osteoblastic phenotypic traits. AB - A clonal cell line, BFO-6, was established in culture from a Dunn osteosarcoma cell line (BFO) and characterized on the basis of bone-inducing activity, alkaline phosphatase activity, PTH sensitive cAMP production and tumorigenicity. Lyophilized pellets of devitalized cells, when implanted into the back muscles of allogenic host mice, consistently elicited heterotopic bone formation at 3 weeks postimplantation. BFO-6 cells were found to be rich in alkaline phosphatase activity and showed a significant response to h-PTH stimulation. The doubling time in the logarithmic phase was 17.2 h and the cells revealed an acrocentric karyotype. Subcutaneous transplantation of cells (1 x 10(7) cells) into a C3H mouse resulted in the production of a tumor with histological features of osteosarcoma. This tumor also retained bone-inducing activity and high alkaline phosphatase activity. PMID- 2553083 TI - The glyoxylate cycle in rat epiphyseal cartilage: the effect of vitamin-D3 on the activity of the enzymes isocitrate lyase and malate synthase. AB - The effect of vitamin-D deficiency and subsequent vitamin-D replacement on the metabolism of rat epiphyseal growth plate cartilage was studied. Biochemical analyses showed the presence of the two unique glyoxylate cycle enzymes isocitrate lyase and malate synthase in cartilage. The activity of these enzymes was markedly increased after treatment with the vitamin. Additionally, rat cartilage showed the capacity to oxidize fatty acid in the presence of cyanide. This cyanide-insensitive fatty acid oxidation is characteristic of peroxisomal B oxidation rather than mitochondrial B-oxidation. Vitamin-D treatment also increased fatty acid oxidation. Lastly, incubation of rat cartilage in the presence of a fatty acid substrate such as palmitate, resulted in a higher tissue glycogen content. Tissue glycogen was further elevated by vitamin-D. Such data indicate the presence of glyoxylate cycle enzymes in a vertebrate tissue and raise the possibility that mammalian cartilage has the capacity to convert lipid to carbohydrate. PMID- 2553084 TI - Effects of pentobarbitone, ketamine and lignocaine on synaptic transmission in the rat olfactory cortex in vitro. AB - The effects of pentobarbitone, ketamine and lignocaine on synaptic transmission in the rat olfactory cortex were studied. All agents depressed the presynaptic spike, the excitatory postsynaptic potential (EPSP) and the population spike evoked by stimulation of the lateral olfactory tract in a concentration-dependent manner. However, each anaesthetic had different effects on the processes of excitatory synaptic transmission. Pentobarbitone depressed primarily the EPSP, ketamine primarily the excitability of the presynaptic fibres and lignocaine the excitability of the presynaptic fibres; lignocaine also reduced the release of excitatory transmitters. None of the three agents influenced the passive membrane properties or the excitability of olfactory pyramidal cells. These data indicate that these agents block synaptic transmission in the olfactory cortex by different mechanisms. PMID- 2553086 TI - Small cell lung cancer cell lines secrete predominantly ACTH precursor peptides not ACTH. AB - A panel of 18 well characterised human small cell lung cancer (SCLC) cell lines was assessed for the production of adrenocorticotrophin (ACTH) and its precursor peptides, pro-opiomelanocortin (POMC) and pro-ACTH. These precursor peptides were measured directly using a novel two-site immunoradiometric assay (IRMA) based on monoclonal antibodies, in conjunction with a similar IRMA for ACTH 1-39. Significant concentrations of ACTH precursors were secreted by 10 of the 18 cell lines (56%). The low levels of ACTH immunoreactivity detected in seven cell lines could be accounted for by the known cross-reactivity of precursors in the ACTH IRMA. This suggests there is little, if any, processing of ACTH precursors to ACTH. Cell pellet extracts contained undetectable or low levels of ACTH precursors and ACTH, indicating that these peptides are not stored intracellularly. During the growth of the SCLC cells in vitro ACTH precursors accumulated progressively in the culture medium. Thus the combination of a direct assay for the ACTH precursors and the panel of SCLC cell lines provides a valuable in vitro model for the expression of POMC in human tumours. PMID- 2553085 TI - Alterations in the production rate and the metabolism of oestrone and oestrone sulphate in breast cancer patients treated with aminoglutethimide. AB - Plasma level, plasma clearance, production rate and interconversions of oestrone and oestrone sulphate were measured in six breast cancer patients receiving aminoglutethimide therapy. Three additional patients had the production rate of oestrone sulphate investigated. Plasma oestrone and oestrone sulphate levels were reduced by a mean of 46% (P less than 0.05) and 71% (P less than 0.005) respectively. These alterations were due to a combined action of aminoglutethimide inhibiting oestrogen production but also increasing oestrogen metabolism. While oestrone and oestrone sulphate production rate was reduced by a mean of 31% (P less than 0.05) and 41% (P less than 0.005) respectively, the plasma clearance rate of oestrone was found to be increased by a mean of 30% (P less than 0.05), and the plasma clearance rate of oestrone sulphate was increased by a mean of 112% during aminoglutethimide treatment. The fraction of oestrone sulphate converted into plasma oestrone was reduced by 52% (P less than 0.05), the transfer of circulating oestrone into sulphate was non-significantly reduced by a mean of 16%. The findings in this investigation show that aminoglutethimide treatment influences oestrogen disposition by mechanisms unrelated to aromatase inhibition. The possibility that such effects might be partly responsible for the mechanism of action of aminoglutethimide in advanced breast cancer should be considered. PMID- 2553087 TI - Loss of chromosome 11p alleles in cultured cells derived from Wilms' tumours. AB - Cell cultures have been produced from five Wilms' tumours. All cultures had a finite lifespan and a pattern of antigen expression which indicated that the cells were derived from the differentiated components of the tumours. No cells showed any of the expected characteristics of the putative Wilms' tumour stem cell. Nevertheless, in both cases where the original tumours showed a loss of heterozygosity at chromosome 11p alleles, the cultured cells also demonstrated a loss of heterozygosity. Thus these cell cultures definitely originated from Wilms' tumour tissue. The results demonstrate that cell cultures can be produced from the differentiated tissues present in Wilms' tumours and that these non immortal cells show no 'transformed' phenotype, even though they possess the genetic changes present in the original tumour. PMID- 2553089 TI - Chemotherapy of non-small cell lung cancer. PMID- 2553088 TI - Bone marrow metastases in small cell lung cancer: detection with magnetic resonance imaging and monoclonal antibodies. AB - The detection of bone marrow involvement might be of prognostic value and may influence therapeutic decisions in small cell lung cancer. By unilateral bone marrow aspiration and biopsy, evidence of bone marrow metastases is seen in 15 30% of patients with this disease. Since magnetic resonance imaging of the lower body and immunostaining with monoclonal antibodies have recently been shown to be very sensitive detection methods, we investigated the value of these two techniques in detecting bone marrow involvement in 35 consecutive patients with small cell lung cancer. The results were compared to those obtained with conventional cytohistological analysis. In all cases when cytology and/or bone marrow biopsy were positive, monoclonal antibodies immunostaining and magnetic resonance imaging also detected malignant cells. Furthermore, evidence of bone marrow involvement was shown with magnetic resonance imaging and/or immunostaining in 10 of 26 cases (38%) where routine procedures were unable to detect malignant cells. In one of these 26 patients, magnetic resonance imaging and immunostaining provided the only evidence of metastatic disease. These data suggest that the rate of bone marrow metastases is underestimated by routine procedures. Further investigation is needed to determine whether or not these new non-invasive methods have prognostic value or affect therapeutic choices in small cell lung carcinoma. PMID- 2553090 TI - Carboplatin, ifosfamide and etoposide with mid-course vincristine and thoracic radiotherapy for 'limited' stage small cell carcinoma of the bronchus. AB - Forty-two patients with small cell lung cancer were treated with a combination of carboplatin, ifosfamide and etoposide. Vincristine was given on day 14 of each course, the courses being repeated every 28 days for a maximum of six. Thoracic radiotherapy was given 4 weeks after the last course of chemotherapy but no prophylactic cranial radiotherapy was administered. Thirty patients had clinically limited state disease, the remaining patients having contralateral neck lymphadenopathy and/or pleural effusions. Elevated enzyme levels (alkaline phosphatase, LDH, ALT, GGT) were noted in 69% of patients. Twenty-four patients (57%) achieved a complete response (CR) when assessed one month after the end of treatment. A further 21% of patients had a partial response (PR). Median duration of CR was 14 months and of PR 8 months. Cerebral metastases were the sole site of relapse in 13% of the CR patients. Myelosuppression was severe with a median nadir of neutropenia of 0.2 x 10(9) cells 1-1. However, 74% of the patient group received all six courses of chemotherapy and only 16 courses (7%) were delayed because of toxicity. There were three deaths associated with treatment-related neutropenia. The median survival of the total group was 14 months, with an actuarial 2 year survival of 37% and a minimum follow-up of 18 months. [A recent analysis, March 1989, demonstrated a 33%, 2 year actual survival.] PMID- 2553091 TI - Radiosensitisation and radioprotection by BSO and WR-2721: the role of oxygenation. AB - Endogenous and exogenous thiols are thought to influence cellular radiosensitivity directly by radical scavenging and/or hydrogen donation processes, and indirectly, by regulating the amount of oxygen (or other electron affinic radiosensitiser) able to reach the radiosensitive targets of the cell. The relative importance of these two mechanisms was evaluated in multicell spheroids treated with two agents currently undergoing clinical testing, the thiophosphate WR-2721 and the glutathione synthesis inhibitor BSO. Fluorescence activated cell sorting techniques were used to recover cells selectively from different depths (different oxygenation status) within the spheroids. The radiosensitivity of cell populations recovered from different regions suggested that both agents acted primarily by affecting the oxygenation status of the spheroid. Similarly, the binding of a fluorescent marker for hypoxic cells, the nitrofuran AF-2, was markedly enhanced by WR-2721 addition, and decreased by BSO induced thiol depletion. We conclude that the major radiobiological consequence of thiol manipulation in multicell systems is to increase or decrease the availability of oxygen. PMID- 2553092 TI - The role of reductive and oxidative metabolism in the toxicity of mitoxantrone, adriamycin and menadione in human liver derived Hep G2 hepatoma cells. AB - The cytotoxic properties of quinones, such as menadione, are mediated through one electron reduction to yield semi-quinone radicals which can subsequently enter redox cycles with molecular oxygen leading to the formation of reactive oxygen radicals. In this study the role of reduction and oxidation in the toxicity of mitoxantrone was studied and its toxicity compared with that of adriamycin and menadione. The acute toxicity of mitoxantrone was not mediated through one electron reduction, since inhibition of the enzymes glutathione reductase and catalase, responsible for protecting the cells against oxidative damage, did not affect its toxicity. Adriamycin was the most potent inhibitor of protein and RNA synthesis of the three quinones. Menadione, at concentrations up to 25 microM, did not inhibit either protein or RNA synthesis unless dicoumarol, an inhibitor of DT-diaphorase, was also present. The two-electron reduction of menadione by DT diaphorase is therefore a protective mechanism in the cell. This enzyme also protected against the toxicity of high concentrations (100 microM) of mitoxantrone. The inhibitory effect of mitoxantrone, but not of menadione or adriamycin, on cell growth was prevented by inhibiting the activity of cytochrome P450-dependent mixed function oxidase (MFO) system using metyrapone. This suggests that mitoxantrone is oxidised to a toxic intermediate by the MFO system. PMID- 2553093 TI - No significant correlation between specific antibodies to mouse mammary tumour virus and human cancer. AB - To study the possible involvement of mouse mammary tumour virus (MMTV) related agent in human cancer we analysed 300 samples of human sera for the presence of antibodies to MMTV structural proteins. All sera were tested by immunoblotting to achieve high specificity. Out of 300 sera, 22 reacted with transframe protein p30, 16 with the ribonucleoprotein p14, six with the envelope glycoprotein gp52 and three with the major core protein p27. Reactivities to p30 and p14 were observed in sera from cancer patients and healthy controls; reactivities to p27 and gp52 predominated in sera of cancer patients. Sera frequently reacted with a 42 kDa protein which is a cellular contaminant of the virus. PMID- 2553094 TI - The prognostic significance of nuclear DNA content in invasive breast cancer--a study with long-term follow-up. AB - The nuclear DNA content of 351 breast carcinomas was determined by flow cytometry from paraffin-embedded tissue to assess the prognostic significance of DNA ploidy, the DNA index (DI) and the S-phase fraction (SPF). The minimum follow-up of the patients was 22 years, and they were all from a defined urban population. DNA ploidy correlated with histological type and grade, mitotic count and nuclear pleomorphism (P less than 0.0001), and also with axillary nodal status (P = 0.0005), tumour necrosis (P = 0.001), primary tumour size (P = 0.03), menopausal status (P = 0.004) and the presence of distant metastases at the time of the diagnosis (P = 0.04). Survival corrected for intercurrent deaths of the patients with a diploid tumour was better than that of the patients with a non-diploid tumour (P = 0.0001, 48% vs 28% at 25 years). SPF had prognostic significance in both axillary node positive and negative patients, but ploidy and DI only in the node negative group, and their significance was greater in post-menopausal than in premenopausal patients. Axillary nodal status, primary tumour size, histological grade and the type of tumour margin circumscription were the most important independent prognostic factors in Cox's multivariate analysis, and SPF had independent prognostic value, whereas ploidy and DI did not. It is concluded that DNA ploidy, DI and SPF have long-term prognostic significance in breast cancer. PMID- 2553095 TI - Rapid diagnosis in varicella and herpes zoster: re-evaluation of direct smear (Tzanck test) and electron microscopy including colloidal gold immuno-electron microscopy in comparison with virus isolation. AB - The Tzanck test and electron microscopy with the technique of colloidal gold labelling in varicella-zoster virus (VZV) infections were compared with virus isolation in 54 patients with clinically suspected varicella or herpes zoster infection. The Tzanck test and direct electron microscopy can determine whether or not an eruption is herpetic but cannot distinguish between herpes simplex virus (HSV) and VZV infection. However, colloidal gold immuno-electron microscopy, using monoclonal antibodies against HSV and anti-VZV IgG, can distinguish between these two herpes viruses. This achieves the same specificity as virus isolation followed by virus neutralization or virus typing using immunofluorescence techniques. The Tzanck test was positive in 91%, virus isolation, under optimal conditions of sampling and transportation, in 80%, direct electron microscopy (negative staining) in 80%, and colloidal gold immuno electron microscopy after a virus concentration procedure in 95% of the cases. The colloidal gold technique offers a rapid diagnosis in patients with suspected VZV infection. PMID- 2553096 TI - Transient erythroblastopenia of childhood due to human parvovirus B19 infection. PMID- 2553097 TI - Virilizing nodular ovarian stromal hyperthecosis, diabetes mellitus and insulin resistance in a postmenopausal woman. Case report. PMID- 2553098 TI - Effect of inhibitors of prostaglandin synthesis on uterine prostaglandin E receptor binding. PMID- 2553099 TI - Interactions of high-density lipoprotein 3 with brain capillary endothelial cells. AB - High-density lipoprotein 3 (HDL3) binds to capillary endothelial cells when their lumen surfaces are exposed to 125I-HDL3 by post-mortem perfusion of whole brain. Kinetic studies of binding of HDL3 to isolated membranes show that HDL3 binds only to endothelial membranes with high affinity (Kd = 7 micrograms/ml). Trypsin treatment of membranes abolishes HDL3 binding. High-affinity binding sites for HDL3 were recovered when endothelial cells from bovine brain capillaries were maintained in culture (Kd = 13 micrograms/ml HDL3 protein). The characteristics of the binding were preserved up to the 6th passage. Competition experiments using isolated luminal membranes or cultured endothelial cells indicate that only HDL3 and not LDL or methylated LDL, are able to compete binding of 125I-HDL3. Furthermore, the inhibition of 125I-HDL3 binding by lipoprotein A-I and lipoprotein A-I:A-II strongly suggests that apolipoprotein A-I is implicated in the formation of HDL3-receptor complexes. The binding is increased by loading cells with free cholesterol or LDL cholesterol. In addition, surface-bound 125I HDL3 remains sensitive to mild trypsin treatment after subsequent incubation of BBCE at 37 degrees C. HDL3 bound to the cell surface is not endocytosed, but rather rapidly released into the medium after binding (t1/2 = 5 min). PMID- 2553100 TI - Interactions of thiophosphatidic acid with enzymes which metabolize phosphatidic acid. Inhibition of phosphatidic acid phosphatase and utilization by CDP diacylglycerol synthase. AB - Thiophosphatidic acid (1,2-diacyl-sn-glycero-3-phosphorothioate; thioPA) was chemically synthesized from egg phosphatidylcholine-derived 1,2-diacylglycerol and PSCl3 and tested for its effects on enzymes which utilize phosphatidic acid (PA) in phospholipid biosynthesis. The compound was not a substrate for rat liver cytosolic PA phosphatase and strongly inhibited this enzyme activity. ThioPA was also a potent inhibitor of purified membrane-associated PA phosphatase from Saccharomyces cerevisiae in a competitive manner and exhibited an apparent Ki = 60 microM. In contrast, purified CDPdiacylglycerol synthase (PA:CTP cytidylyltransferase) from this organism was able to convert thioPA to CDP diacylglycerol. The apparent Vmax for thioPA was 7-fold lower than that for PA, whereas the apparent Km for thioPA (70 microM) was 4-fold lower than that for PA. Calculation of the specificity constant (Vmax/Km) demonstrated that PA was the preferred substrate. These properties of thioPA indicate that this substance may prove useful in studies of phospholipid metabolism and function. PMID- 2553101 TI - Down-regulation of histone H3 and H4 gene transcription in differentiated L6 myotubes. AB - The core histone mRNA levels in terminally differentiated L6 myotubes decrease to less than 5% of the amount present in proliferating myoblasts in parallel with the cessation of DNA synthesis (Bird, RC., Jacobs, F.A., Stein, G., Stein, J. and Sells, B.H. (1985) Biochim. Biophys. Acta 824, 209-217). The role of gene transcription in the down-shift of histone mRNA levels was assessed using a cell free system. The level of transcription from the differentiation-independent adenovirus major late promoter was directly related to the RNA polymerase II activity of myoblast and myotube nuclear extracts. In addition, both extracts actively transcribed the histone H4 gene template containing only the 5 proximal promoter region (-210 bp). In contrast, inclusion of the distal-proximal promoter region (-410 to -210 bp) in the template resulted in a 60% decrease in transcription by the myotube extract. A similar down-shift in transcription of the histone H3 gene template (containing 900 bp 5 of the initiation site) by myotube nuclear extracts was also observed. The decrease in histone mRNA levels in myotubes may therefore be controlled in part by a transcriptional mechanism involving a negative regulatory factor. PMID- 2553102 TI - Enhancement of c-erbA proto-oncogene expression by glucocorticoid hormones in S49.1 lymphoma cells. AB - The modifications of the mRNA levels of the c-myc and c-erbA proto-oncogenes during the dexamethasone-induced decrease of S49.1 cell proliferation have been studied. The levels of c-myc mRNA decreased significantly between 3 and 18 h after dexamethasone (1 microM) treatment. In contrast, a significant increase in the levels of a 2.6 kb c-erbA mRNA was observed between 6 and 18 h after hormone treatment. Cycloheximide treatment of S49.1 cells increased the levels of c-erbA RNA and overcome the enhancing effect of dexamethasone on the expression of this proto-oncogene, suggesting that ongoing protein synthesis is necessary to elicit this hormone effect. The associated decrease of cell proliferation and changes in c-myc and c-erbA mRNA levels after dexamethasone treatment suggest that such oncogenes might be involved in the dexamethasone-mediated control of lymphoid cell growth. PMID- 2553103 TI - Inhibition of the liver cell receptor-activated Ca2+ inflow system by metal ion inhibitors of voltage-operated Ca2+ channels but not by other inhibitors of Ca2+ inflow. AB - The properties of the receptor-activated Ca2+ inflow system in the liver cell plasma membrane were compared with those of voltage-operated Ca2+ channels and receptor-operated Ca2+ channels present in other cell types by testing the susceptibility of the Ca2+ inflow system to inhibition by other metal ions and known inhibitors of Ca2+ movement across membranes. Co2+ inhibited Ca2+ inflow through the receptor-activated Ca2+ inflow system, as assessed by measurement of (a) the activation by extracellular Ca2+ (Cao2+) of glycogen phosphorylase in the presence of vasopressin and (b) 45Ca2+ exchange in the presence of the hormone. The concentration of Co2+ which gave half-maximal inhibition was 280 microM. The inhibition by Co2+ was reversed by high Cao2+. Co2+ did not inhibit basal Ca2+ inflow as measured by 45Ca2+ exchange in the absence of vasopressin. Zn2+, Cd2+, Ni2+ and Mn2+ each inhibited Ca2+ inflow through the receptor-activated Ca2+ inflow system. The concentrations of these ions which gave half-maximal inhibition were 10, 50, 220 and 400 microM, respectively. Little inhibition of receptor-activated Ca2+ inflow was observed in the presence of Sr2+ or Ba2+. However, substantial amounts of 90Sr2+ were taken up by hepatocytes. Rates of 90Sr2+ uptake increased from 0.5-8 nmol per min per mg wet wt. when the extracellular concentration of Sr2+ was varied from 0.25 to 2.5 mM. Sr2+ uptake was inhibited 50% by Cao2+ with half-maximal inhibition at 100 microM Cao2+, but was not inhibited by verapamil and was not stimulated by vasopressin. The movement of Ca2+ through the receptor-activated Ca2+ inflow system was not inhibited by high concentrations of each of a number of inhibitors of voltage operated and receptor-operated Ca2+ channels and intracellular Ca2+ movement. It is concluded that while the susceptibility to inhibition by metal ions of the receptor-activated Ca2+ inflow system in the liver cell plasma membrane is similar to that of voltage-operated Ca2+ channels, there are significant differences between the liver cell receptor-activated Ca2+ inflow system and both voltage-operated Ca2+ channels and some other receptor-operated Ca2+ channels with respect to inhibition by organic compounds. PMID- 2553104 TI - Fatty acid uptake by human hepatoma cell lines represents a carrier-mediated uptake process. AB - Cellular influx kinetics of a representative long chain fatty acid, [3H]oleate, were examined in monolayer cultures of three different human hepatoma cell lines (Hep G2; PLC/PRF 5; Mz-Hep-1). The cultures were incubated with 173 microM [3H]oleate in the presence of various concentrations of albumin which served to modulate the unbound oleate concentration in the medium. For all [3H]oleate albumin complexes incubated, it was shown that cellular uptake of [3H]oleate over the initial 30 s incubation period was maximal, linear and independent of intracellular fatty acid metabolism, representing cellular influx. With increasing unbound oleate concentrations in the medium cellular influx by all three cell lines revealed similar saturation kinetics with Km values of 112.6 +/- 14.5 nM and Vmax values of 7.19 +/- 0.32 nmol.min-1 per mg cell protein. When these hepatoma cell lines were pretreated with the IgG fraction of a monospecific antibody to the rat liver membrane fatty acid binding protein (MFABP), initial uptake of [3H]oleate was selectively inhibited compared to controls pretreated with the IgG fraction of the preimmune serum. Furthermore, immunoblot analysis with the monospecific antibody to the rat MFABP revealed reactivity with a single 40 kDa protein in the homogenates of all three cell lines. These data suggest that uptake of fatty acids by human hepatoma cells may be mediated by a specific membrane fatty acid binding protein. PMID- 2553105 TI - Carcinoma autoantigens T and Tn and their cleavage products interact with Gal/GalNAc-specific receptors on rat Kupffer cells and hepatocytes. AB - We studied interactions of isolated Thomsen-Friedenreich (T)- and Tn-specific glycoproteins with the Gal/GalNAc-specific receptors on rat Kupffer cells and compared them to those with rat hepatocytes. Immunoreactive T and Tn are specific pancarcinoma epitopes. Electron microscopy of gold-labelled T and Tn antigens revealed their specific binding to Kupffer cells, followed by their uptake via the coated pit/vesicle pathway of receptor-mediated endocytosis. Preincubation of Kupffer cells with GalNAc and GalNAc-BSA, but not GlcNAc or GlcNAc-BSA specifically inhibited binding of the T and Tn glycoproteins. Desialylated, isologous erythrocytes (T RBC) are known to bind to the Gal/GalNAc receptors of rat Kupffer cells and hepatocytes. This attachment was specifically inhibited by T and Tn in a concentration-dependent manner: 50% T RBC-Kupffer cell contacts were inhibited at 8.5.10(-6) mM T and 8.5.10(-5) mM Tn antigen concentrations, respectively. The corresponding figures for hepatocytes were 6.10(-6) mM T and 1.2.10(-6) mM Tn antigen. Amino-terminal cleavage products of the T glycoprotein, possessing clusters terminating in non-reducing Gal/GalNAc, inhibited T RBC binding to Kupffer cells and hepatocytes usually at 10(-2) to 10(-5) mM concentrations, whereas GalNAc, galactose and galactose glycosides inhibited at millimolar concentrations. Galactose-unrelated carbohydrates were inactive at concentrations greater than or equal to 50 mM. PMID- 2553106 TI - Metabolic effects of forskolin in chick chondrocytes. AB - The effects of forskolin on parameters of energy metabolism and proteoglycan synthesis have been investigated in chick embryo sternal chondrocyte cultures. After 8 h exposure to 100 microM forskolin, ATP levels and oxygen consumption were unaltered. Protein synthesis was unaffected up to 50 microM forskolin and protein degradation was unaffected by forskolin up to 100 microM. In contrast, incorporation of the proteoglycan precursors, 35SO4 and [3H]glucosamine, was more sensitive to forskolin. Inhibition was linear with dose between 10 and 100 microM, reaching 70% at 100 microM. Incorporation of 35SO4 into glycosaminoglycan chains initiated on an artificial beta-xyloside acceptor was inhibited in the same manner. cAMP accumulation was maximal at 10 microM forskolin, a concentration which did not alter proteoglycan synthesis. We conclude that a major, acute effect of forskolin in these short-term experiments is inhibition of proteoglycan synthesis in a cAMP-independent manner. PMID- 2553107 TI - Purification and partial characterization of protein phosphatases from rat thymus. AB - Protein phosphatases assayed with phosphorylase alpha are present in the soluble and particulate fractions of rat thymocytes. Phosphorylase phosphatase activity in the cytosol fraction was resolved by heparin-Sepharose chromatography into type-1 and type-2A enzymes. Similarities between thymocyte and muscle or liver protein phosphatase-1 included preferential dephosphorylation of the beta subunit of phosphorylase kinase, inhibition by inhibitor-2 and retention by heparin Sepharose. Similarities between thymocyte and muscle or liver protein phosphatase 2A included specificity for the alpha subunit of phosphorylase kinase, insensitivity to the action of inhibitor-2, lack of retention by heparin Sepharose and stimulation by polycationic macromolecules such as polybrene, protamine and histone H1. Protein phosphatase-1 from the cytosol fraction of thymocytes had an apparent molecular mass of 120 kDa as determined by gel filtration. The phosphatase-2A separated from the cytosol of thymocytes may correspond to phosphatase-2A0, since it was completely inactive (latent) in the absence of polycation and had activity only in the presence of polycations. The apparent molecular mass of phosphatase-2A0 from thymocytes was 240 kDa as determined by gel filtration. The catalytic subunit of thymocyte type-1 protein phosphatase was purified with heparin-Sepharose chromatography followed by gel filtration and fast protein liquid chromatography on Mono Q column. The purified type-1 catalytic subunit exhibited a specific activity of 8.2 U/mg and consisted of a single protein of 35 kDa as judged by SDS-gel electrophoresis. The catalytic subunit of type-2A phosphatase from thymocytes appearing in the heparin-Sepharose flow-through fraction was further purified on protamine-Sepharose, followed by gel filtration. The specific activity of the type-2A catalytic subunit was 2.1 U/mg and consisted of a major protein of 34.5 kDa, as revealed by SDS-gel electrophoresis. PMID- 2553108 TI - Characterization of subclones of Madin-Darby canine kidney renal epithelial cell line. AB - Heterogeneity in Madin-Darby canine kidney (MDCK) epithelial cells has been reported, however, its details have not been well described. In the present study, we show that subclones obtained from a MDCK cell line could be divided into two morphologically and biochemically distinct cell types with different hormonal responsiveness. Clones of the first type, motile clones, which had extended and flattened cytoplasm, were devoid of carbonic anhydrase activity. Clones of the second type, nonmotile clones, formed colonies of cuboidal cells and showed carbonic anhydrase activity. Motile clones synthesized cAMP in response to arginine vasopressin, prostaglandin E1, and isoproterenol but not glucagon. In contrast, nonmotile clones responded to all of these hormones. These findings suggest MDCK cells have multiple cellular origins. The motile clones have characteristics similar to the principal cells of the collecting system, whereas the nonmotile clones may be derived from the thick ascending limb or the intercalated cell. Our studies also demonstrate a significant influence of culture condition on MDCK cellular behavior (carbonic anhydrase activity, Na+/K+ ATPase activity and vasopressin responsiveness). Therefore, physiologic and biochemical experiments with MDCK cells must be interpreted with reservations about cellular heterogeneity as well as differences induced by culture conditions. PMID- 2553109 TI - Evidence for an alteration in the microsomal Ca2+ release mechanism in senescent rat parotid acinar cells. AB - Previously, we have shown that Ca2+ mobilization following an alpha 1-adrenergic receptor stimulus is reduced in parotid acinar cells from senescent rats as a result of an altered ability of inositol 1,4,5-trisphosphate (IP3) to induce Ca2+ release from a non-mitochondrial, intracellular Ca2+ store (Ishikawa, Y., et al. Biochim. Biophys. Acta 968, 203-210). We have used this model to examine the IP3 induced Ca2+ release mechanism in these cells. 45Ca2+ efflux, after exposure to ( ) epinephrine, from cells of young adult (3-6 months) rats was approx. 2-fold that observed from cells from older animals (approx. 24 months) either in the presence or absence of extracellular Ca2+. Similarly, cytosolic Ca2+ levels were greater in cells of young adult rats under these same incubation conditions. However, microsomal membrane preparations, from both age groups displayed similar IP3 binding sites (Kd approximately 90 nM, Bmax approximately 850 fmol/mg protein) and ATP-dependent Ca2+ transport ability (approx. 8 nmol/mg protein.min 1). These data suggest that there is an alteration in the IP3-induced Ca2+ release mechanism in microsomal membranes of parotid glands from senescent rats which may account for the decreased Ca2+ release seen after agonist stimulation of this tissue. PMID- 2553110 TI - Studies on the mechanism by which prolactin regulates protein, RNA, and DNA synthesis in Nb2 node lymphoma cells. AB - Specific aspects of the prolactin stimulation of RNA, DNA and protein synthesis in the Nb2 node lymphoma cell line were determined. In time sequence studies the onset of the prolactin stimulation of the incorporation of radiolabeled precursors into these macromolecules was found to be 0.5-1 h for [3H]uridine incorporation into RNA, 1-2 h for [3H]leucine incorporation into protein, and 4-8 h for [3H]thymidine incorporation into DNA. The total DNA content of the cell cultures was increased by 12-18 hours after addition of prolactin. Amiloride, an inhibitor of the plasma-membrane-bound Na+/H+ antiporter, was found to inhibit the mitogenic effects of prolactin. Amiloride was also found to inhibit the prolactin stimulation of DNA, RNA and protein synthesis, thus suggesting that the initial regulation of the Na+/H+ antiporter may initiate these responses as well as the mitogenic effect of prolactin. In contrast, H-7, a drug which inhibits protein kinase C, had no effect on the magnitude of the prolactin stimulation of DNA, RNA or protein synthesis at a drug concentration (100 muM) that abolished the mitogenic effect of prolactin. The early effects of prolactin on RNA, DNA and protein synthesis would therefore appear not to involve an activation of protein kinase C. PMID- 2553111 TI - Slip and leak in mitochondrial oxidative phosphorylation. AB - During oxidative phosphorylation by mammalian mitochondria part of the free energy stored in reduced substrates is dissipated and energy is released as heat. Here I review the mechanisms and the physiological significance of this phenomenon. PMID- 2553112 TI - ENDOR studies of the intermediate electron acceptor radical anion I-. in Photosystem II reaction centers. AB - The EPR and ENDOR characteristics of the intermediate electron acceptor radical anion I-. in Photosystem II (PS II) are shown to be identical in membrane particles and in the D1D2 cytochrome b-559 complex (Nanba, O. and Satoh, K. (1987) Proc. Natl. Acad. Sci. USA 84, 109-112). These findings provide further evidence that the D1D2 complex is the reaction center of PS II and show that the pheophytin binding site is intact. A hydrogen bond between I-. and the protein (GLU D1-130) is postulated on the basis of D2O exchange experiments. The ENDOR data of I-. and of the pheophytin a radical anion in different organic solvents are compared and the observed differences are related to structural changes of the molecule on the basis of molecular orbital calculations (RHF-INDO/SP). The importance of the orientation of the vinyl group (attached to ring I) on electron transfer is discussed. PMID- 2553113 TI - Hyperosmotic relaxation lysis of chromaffin granules is caused by interactions between the granular membrane and intragranular vesicles. AB - Bovine chromaffin granules undergo irreversible structural changes during osmotic shrinkage in hypertonic sucrose and salt solutions, such that, on reexposure to isoosmotic conditions they do not regain their original morphology, but undergo lysis ('hyperosmotic relaxation lysis'). Irreversible alterations of granules were induced by hypertonic incubations lasting for as little as 1 min. Fluorescence and EPR membrane labelling experiments showed that hypertonicity did not induce membrane loss for instance by inwardly or outwardly directed pinching off of membrane material. The mean sizes of chromaffin granules as a function of increasing and subsequently decreasing osmotic pressure were measured by photon correlation spectroscopy; there was no significant difference in sizes of hyperosmotically pretreated granules as compared with controls. Freeze-fracture electron micrographs showed the formation of 'twins' and 'triplets' under hypertonic conditions. They also revealed intragranular vesicles of 50-200 nm in diameter in both hypertonically and isotonically suspended granules. 'Twin' and 'triplet' granules were formed by the attachment of intragranular vesicles to the granule membranes. We suggest that hyperosmotic relaxation lysis is caused by the fact that this adhesion partly prevents the granule membrane from reexpanding, thus, leading to its rupture. PMID- 2553114 TI - ESR study on synthetic glyceroglycolipid liposomal membranes. AB - We previously reported that glyceroglycolipid liposomes without cholesterol activated mouse peritoneal macrophages in vivo and in vitro, whereas glyceroglycolipid liposomes containing equimolar cholesterol did not. In order to characterize the properties of the glyceroglycolipid membranes, ESR spectroscopic studies were carried out with an acyl spin-labeled galactosyl ceramide (SL-GC) or a headgroup spin-labeled phospholipid (SL-6-DPPA) in 1,2-dipalmitoyl[beta cellobiosyl-(1'---3)]glycerol (Cel-DAG) liposomal membranes. The ESR spectrum of the SL-GC in the Cel-DAG liposomes at 37 degrees C was a single broad line, indicating that the SL-GC molecules were excluded almost completely from Cel-DAG domains and formed clusters in the membranes. The spectrum of SL-6-DPPA in the Cel-DAG liposomes at 37 degrees C showed broad resonance lines with the central peak being the highest, while that at 60 degrees gave narrow lines with the low field peak being the highest. This observation and rotational correlation time analysis showed that the molecular motions of spin-label moiety of the SL-6-DPPA were extremely restricted at 37 degrees C but not above Tc. These results suggest that below Tc the Cel-DAG molecules are packed tightly and restricted in motion in the membrane. Incorporation of cholesterol into the Cel-DAG liposomal membranes gave (1) the spectra of the SL-GC triplet, and (2) the spectra of the SL-6-DPPA narrow resonance with the low-field peak being the highest. These results suggest that cholesterol disturbs the rigid-packed structure of the Cel DAG membrane and increases the molecular motions of the Cel-DAG. The DSC analysis of Cel-DAG with and without cholesterol agreed well to the results of the ESR technique. Thus we assume that peritoneal macrophages recognize the rigid-packed carbohydrate residues which are restricted in motion on the Cel-DAG membranes. PMID- 2553115 TI - Use-dependent block of sodium channels in frog myelinated nerve by tetrodotoxin and saxitoxin at negative holding potentials. AB - Na+ currents were measured in myelinated frog nerve fibres in the presence of nanomolar concentrations of tetrodotoxin (TTX) or saxitoxin (STX) in the extracellular solution. The Na+ currents declined during a train of depolarizing pulses if the fibre was held at hyperpolarizing potentials between the pulses. At a pulse frequency of 0.8 Hz, the peak Na+ currents were reduced to 70 or 60% of the initial value in 9.3 nM TTX and 3.5 nM STX solutions, respectively. A decline of Na+ currents was also observed in two-pulse experiments. The peak Na+ current during a second test pulse did not depend on the duration (0.2 to 12 ms) of the first pulse. It decreased with increasing interval between the pulses, reached a minimum and increased again. The results are interpreted with a use-dependent blockage of Na+ channels by TTX or STX at negative holding potentials. The effects were described quantitatively, assuming a fast affinity increase of toxin receptors at Na+ channels triggered by Na+ activation followed by slow toxin binding to channels and relaxation of the receptor affinity. PMID- 2553116 TI - Binding of tetrodotoxin and saxitoxin to Na+ channels at different holding potentials: fluctuation measurements in frog myelinated nerve. AB - The number of available Na+ channels in nodes of frog nerve fibres was determined from nonstationary Na+ current fluctuations recorded during a train of depolarizing test pulses. Mean numbers in Ringers's solution were 90,000 at a hyperpolarizing holding potential VH = -40 mV, 50,000 at the resting potential (VH = 0 mV) and 30,000 at a depolarizing holding potential VH = 30 mV. Addition of the cationic channel blockers tetrodotoxin (TTX) or saxitoxin (STX) to Ringer reduced the channel number by a factor which was independent of the holding potential. The reduction factor was 4 for 9.3 nM TTX and 3 for 3.5 nM STX. Thus, in the state of repetitive stimulation, TTX or STX blockage of Na+ channels is hardly affected by the membrane potential. Taking into account use-dependent TTX and STX effects [1], it is concluded that binding of both toxins exhibits a weak voltage dependence with toxin affinities decreasing at more negative holding potentials. The results suggest that binding of TTX and STX occurs at an external superficial receptor near the Na+ channel and that the toxin affinity of the receptor may be modulated by fast Na+ activation and slow inactivation gating processes. PMID- 2553117 TI - Use of spin labels to determine the percentage of interdigitated lipid in complexes with polymyxin B and polymyxin B nonapeptide. AB - Long chain spin labels with the nitroxide group located near the terminal methyl of the chain were used to determine the percentage interdigitated lipid in complexes of polymyxin B (PMB) and polymyxin B nonapeptide (PMBN) with the acidic lipids dipalmitoylphosphatidylglycerol (DPPG) and dipalmitoylphosphatidic acid (DPPA) at varying mole ratios of drug to lipid and at different pH values. These spin labels are more motionally restricted in the interdigitated than in the non interdigitated gel phase bilayer. This allows determination of the percentage interdigitated lipid by resolution of the spectrum into motionally restricted and more mobile components. At nonsaturating concentrations of PMB, significantly more DPPG than that which can be maximally PMB-bound, becomes interdigitated. As the temperature approaches the gel to liquid crystalline phase transition temperature, the bilayer becomes progressively non-interdigitated. The ESR spectrum indicates that PMB also causes interdigitation of DPPA. However, in contrast to DPPG, the amount of DPPA which is interdigitated at pH 6, is less than the amount which is expected to be PMB-bound. This is attributed to the ability of DPPA to participate in lateral interlipid hydrogen bonding interactions. Such lateral interactions would be abolished in the interdigitated bilayer and thus they are expected to inhibit its formation. At pH 9, where the interlipid interactions of DPPA are weakened, PMB induces even more lipid than that which is PMB-bound to become interdigitated. Indeed, the percentage interdigitated lipid is even greater than found for DPPG. This may be partly a result of the greater negative charge of DPPA at this pH. A greater repulsive negative charge is expected to favor interdigitation. PMBN is less effective than PMB at inducing interdigitation of DPPG and causes little or no interdigitation of DPPA at pH 6, even at saturating concentrations. PMBN also does not lower the phase transition temperature of DPPA at pH 6 as much as PMB. At pH 9, the effect of PMBN on DPPA is more similar to the effect of PMB. However, even for DPPG, and DPPA at pH 9, PMBN does not maintain interdigitation of the lipids at higher temperatures as effectively as PMB. PMBN's smaller perturbing effect and greatly decreased ability to cause interdigitation of DPPA at pH values below 9 may be related to a decreased ability to cause lateral separation of the lipid molecules, which is necessary in order to weaken the interlipid interactions.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2553118 TI - Interdigitation of phosphatidylcholine and phosphatidylethanolamine mixed with complexes of acidic lipids and polymyxin B or polymyxin B nonapeptide. AB - A fatty acid spin label, 16-doxyl-stearic acid, was used to determine the percent interdigitated lipid in mixtures of a neutral phospholipid and an acidic phospholipid. Interdigitation of the acidic lipid was induced with polymyxin B (PMB) at a mole ratio of PMB to acidic lipid of 1:5. This compound does not bind significantly to neutral lipids or induce interdigitation of the neutral lipids by themselves. The neutral lipids used were dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), or dipalmitoylphosphatidylethanolamine (DPPE), and the acidic lipids were dipalmitoylphosphatidylglycerol (DPPG) or dipalmitoylphosphatidic acid (DPPA). The percent interdigitated lipid was determined from the percent of the spin label which is motionally restricted, assuming that the spin label is homogeneously distributed in the lipid. Assuming further that 100% of the acidic lipid is interdigitated at this saturating concentration of PMB, the percentage of the neutral lipid which can become interdigitated along with it was calculated. The results indicate that about 20 mole % DPPC can be incorporated into and become interdigitated in the interdigitated bilayer of PMB/DPPG at 4 degrees C. As the temperature approaches the phase transition temperature, the lipid becomes progressively less interdigitated; this occurs to a greater degree for the mixtures than for the single acidic lipid. Thus the presence of DPPC promotes transformation of the acidic lipid to a non-interdigitated bilayer at higher temperatures. At the temperature of the lipid phase transition little or none of the lipid in the mixture is interdigitated. Thus the lipid phase transition detected by calorimetry is not that of the interdigitated bilayer. The shorter chain length DMPC can be incorporated to a greater extent than DPPC, 30 50 mol%, in the interdigitated bilayer of PMB-DPPG. This may be a result of reduced exposure of the terminal methyl groups of the shorter myristoyl chains at the polar/apolar interface of the interdigitated bilayer. Less than 29% of the total lipid was interdigitated in a DPPC/DPPA/PMB 1:1:0.2 mixture indicating that none of the DPPC in this mixture becomes interdigitated. This is attributed to the lateral interlipid hydrogen bonding interactions of DPPA which inhibits formation of an interdigitated bilayer. DPPE was found to be incorporated into the interdigitated bilayer of PMB-DPPG to a similar extent as DPPC if the amount of PMB added is sufficient to bind to only the DPPG in the mixture. Differential scanning calorimetry showed that the remaining non-interdigitated DPPE-enriched mixture phase separates into its own domain.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2553119 TI - Thallium binding to native and radiation-inactivated Na+/K+-ATPase. AB - The number of high-affinity K+-binding sites on purified Na+/K+-ATPase from pig kidney outer medulla has been assessed by measurement of equilibrium binding of thallous thallium, Tl+, under conditions (low ionic strength, absence of Na+ and Tris+) where the enzyme is in the E2-form. Na+/K+-ATPase has two identical Tl+ sites per ADP site, and the dissociation constant varies between 2 and 9 microM. These values are identical to those for Tl+ occlusion found previously by us, indicating that all high-affinity binding leads to occlusion. The specific binding was obtained after subtraction of a separately characterized unspecific adsorption of Tl+ to the enzyme preparations. Radiation inactivation leads to formation of modified peptides having two Tl+-binding sites with positive cooperativity, the second site-dissociation constant approximating that for the native sites. The radiation inactivation size (RIS) for total, specific Tl+ binding is 71 kDa, and the RIS for Tl+ binding with original affinity is approx. 190 kDa, equal to that of Na+/K+-ATPase activity and to that for Tl+ occlusion with native affinity. This latter RIS value confirms our recent theory that in situ the two catalytic peptides of Na+/K+-ATPase are closely associated. The 71 kDa value obtained for total Tl+ sites is equal to that for total binding of ATP and ADP and it is clearly smaller than the molecular mass of one catalytic subunit (112 kDa). The Tl+-binding experiments reported thus supports the notion that radiation inactivation of Na+/K+-ATPase is a stepwise rather than an all or none process. PMID- 2553120 TI - The ontogeny of rat gastric H+/K+-ATPase. AB - The ontogeny of rat H+/K+-ATPase was studied between foetal day 18 and neonatal day 18, using a specific monoclonal antibody (95-111 mAb). The H+/K+-ATPase content of gastric subcellular membranes was assayed and the ATPase subunits were characterized by Western blot. The epithelium density in parietal cells was measured by immunohistochemistry. H+/K+-ATPase was present in the 18-day-old foetuses and parietal cells were detected on foetal day 19. The H+/K+-ATPase concentration remained stable from foetal day 18 to neonatal day 1, while the parietal cell density increased 2.5-fold. The H+/K+-ATPase concentration increased by 2.5-fold on day 6, then remained constant up to day 18. The parietal cell density remained unchanged during this period, suggesting that the concentration increase on day 6 was due to an increase in parietal cell ATPase content. The 95-111 mAb recognized a 95 kDa single band on foetal day 18 and a doublet at all the other stages of development. Previous studies had demonstrated that acid secretion drops critically at day 12 post partum in the rat and that H+/K+-ATPase activity is lost. The present study demonstrates that the H+/K+ ATPase is, however, present on day 12. PMID- 2553121 TI - Identification, partial purification and characterization of high-molecular weight gelatin-degrading metalloproteinases produced by a rat mammary carcinoma cell line. AB - BC1 rat mammary carcinoma cells were found to secrete a unique profile of metalloproteinases, distinguished by two gelatin-degrading metalloproteinases of Mr greater than 220.10(3) and Mr much greater than 220.10(3). These enzymes were each partially purified by gel-filtration chromatography, and inhibitor studies showed them to be metalloproteinases. Under conditions where denatured collagen types I, II, and V were completely degraded, native collagen types I, II, IV and V, fibronectin, fibrinogen, C1q, casein, and denatured transferrin were not degraded significantly by these enzymes. The relationship of these enzymes to other extracellular matrix-degrading metalloproteinases and their possible significance in tumour invasion and metastasis is discussed. PMID- 2553122 TI - Chicken skeletal muscle has three Ca2+-dependent proteinases. AB - Chicken breast muscle has three Ca2+-dependent proteinases, two requiring millimolar Ca2+ (m-calpain and high m-calpain) and one requiring micromolar Ca2+ (mu-calpain). High m-calpain co-purifies with mu-calpain through successive DEAE cellulose (steep gradient), phenyl-Sepharose, octylamine agarose, and Sephacryl S 300 columns, but elutes after mu-calpain when using a shallow KCl gradient to elute a DEAE-cellulose column. The mu- and m-calpains have 80 and 28 kDa polypeptides and are analogous to the mu- and m-calpains that have been purified from bovine, porcine and rabbit skeletal muscle. High m-calpain, which seems to be a new Ca2+-dependent proteinase, is still heterogeneous after the DEAE cellulose column eluted with a shallow KCl gradient. Additional purification through two successive HPLC-DEAE columns and one HPLC-SW-4000 gel permeation column produces a fraction having six major polypeptides and 6-8 minor polypeptides on SDS-PAGE. A 74-76 kDa polypeptide in this fraction reacts in Western blots with monospecific, polyclonal anti-calpain antibodies that react with both the 80 kDa and the 28 kDa polypeptides of mu- or m-calpain. High m calpain also is related to mu- and m-calpain in that it causes the same limited digestion of skeletal muscle myofibrils, has a similar pH optimum near pH 7.9 8.4, requires Ca2+ for activity, and reacts with the calpain inhibitor, calpastatin, and a variety of serine and cysteine proteinase inhibitors in a manner identical to mu- and m-calpain. High m-calpain differs from mu- and m calpain in its elution off DEAE-cellulose columns and its requirement of 3800 microM Ca2+ for one-half maximal activity compared with 5.35 microM Ca2+ for mu calpain and 420 microM Ca2+ for m-calpain. The physiological significance of high m-calpain in unclear. The presence of mu-calpain in chicken breast muscle suggests that all skeletal muscles contain both mu- and m-calpain, although the relative proportions of these two proteinases may vary in different species. PMID- 2553123 TI - A naturally occurring molecular form of human plasma cholinesterase is an albumin conjugate. AB - Human plasma cholinesterase (acylcholine acylhydrolase, EC 3.1.1.8) consists of four main molecular forms designated as C1, C2, C3 and C4 according to their electrophoretic mobility on gels. The major component, C4, is the tetrameric form; C1 and C3 are the monomeric and dimeric forms, respectively. The C2 form, which has an apparent free electrophoretic mobility higher than that of the three size isomers, and, moreover, a higher isoelectric point, was found to be a covalent conjugate between the cholinesterase monomer and serum albumin. This result is supported by the following arguments: the non-catalytic subunit of C2 was found to be a carbohydrate-free protein of apparent molecular mass 65 kDa that could not be labelled by diisopropylfluorophosphonate in the labelling conditions of esterases. It possesses a high affinity for a long-chain aliphatic ligand (a substituted octadecylamine) and for Cibacron blue F3 GA, and could be adsorbed on an immunoadsorbent for albumin. The two subunits of C2 are disulfide bridge linked; the active center of the cholinesterase subunit is partly masked by the albumin molecule. The conjugation reaction very likely occurs in the hepatic cell and not in plasma. PMID- 2553124 TI - The use of folding patterns in a multisolution approach of the all-beta protein three-dimensional structure. A beta-roll structure is predicted in the retroviral glycoprotein. AB - An automatic macromolecular modelling package of unknown protein structures was developed using the intimate correlation which appears between the observed X-ray structures and their associated predicted folding patterns. The method can be considered as a generalization of both the combinatorial [1] and the template identification [2,3] approaches which were proposed some years ago, and provide a fast way of selecting 'structural motifs' to build new proteins. As an illustration, the tertiary fold of the all-beta-domain of the retroviral outermembrane glycoprotein is proposed. PMID- 2553125 TI - Glycolytic enzyme interactions with tubulin and microtubules. AB - Interactions of the glycolytic enzymes glucose-6-phosphate isomerase, aldolase, glyceraldehyde-3-phosphate dehydrogenase, triose-phosphate isomerase, enolase, phosphoglycerate mutase, phosphoglycerate kinase, pyruvate kinase, lactate dehydrogenase type-M, and lactate dehydrogenase type-H with tubulin and microtubules were studied. Lactate dehydrogenase type-M, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase, and aldolase demonstrated the greatest amount of co-pelleting with microtubules. The presence of 7% poly(ethylene glycol) increased co-pelleting of the latter four enzymes and two other enzymes, glucose-6-phosphate isomerase, and phosphoglycerate kinase with microtubules. Interactions also were characterized by fluorescence anisotropy. Since the KD values of glyceraldehyde-3-phosphate dehydrogenase, pyruvate kinase and lactate dehydrogenase for tubulin and microtubules were all found to be between 1 and 4 microM, which is in the range of enzyme concentration in cells, these enzymes are probably bound to microtubules in vivo. These observations indicate that interactions of cytosolic proteins, such as the glycolytic enzymes, with cytoskeletal components, such as microtubules, may play a structural role in the formation of the microtrabecular lattice. PMID- 2553126 TI - The relationship between the glucose oxidase subunit structure and its thermostability. AB - The thermostability of glucose oxidase (beta-D-glucose: oxygen 1-oxidoreductase, EC 1.1.3.4) at 60 degrees C has been studied as a function of its concentration in various media (pure water and pure deuterium oxide). In deuterium oxide, glucose oxidase is more stable than in water, and two kinds of stabilizing effect have been observed: the medium-organization effect and the enzyme-concentration effect. This effect has been related to the glucose oxidase subunit structure. This enzyme contains four forms of subunit: monomer, dimer, trimer, and tetramer, which are all composed of the identical monomer. The monomers of glucose oxidase subunits are linked by the non-covalent bond. Only dimer and trimer possess the enzymatic activity. During glucose oxidase denaturing, monomers assemble into dimer, trimer, or tetramer. This redistribution behavior depends on the enzyme concentration and the nature of the medium. PMID- 2553127 TI - [Adsorption of fibrinogen on silicon oxide with controlled hydrophobic/hydrophilic properties]. AB - The amount of fibrinogen irreversibly adsorbed on silicon dioxide does not exceed 3.6 pmol/cm2 and depends on the protein concentration, solution pH and surface hydrophobic/hydrophilic properties. Electrostatic interactions determine the fibrinogen adsorption rate. Partial denaturation of fibrinogen takes place in its adsorption form diluted solutions with the pH value lower than the protein isoelectric point. PMID- 2553128 TI - [Inactivation of alpha VEE virus using a high intensity laser with UV-radiation]. AB - Inactivation of VEE virus with laser UV impulses of nano- and picosecond duration was investigated. It has been shown that in both cases there is a decrease of the inactivation cross-section with the rise of irradiation intensity. It points to the fact that the major lethal photoproduct in VEE is formed by a single-quantum mechanism. PMID- 2553129 TI - [Regularity of changes in magnetic characteristics of Saccharomyces cerevisiae yeast cells at various stages of culture growth]. AB - Correlation between cell cycle of the synchronous yeast culture and ESR signal intensity at g 2.2 and 77 K was studied. It was shown that the maximal intensity of ESR signal was reached 10-15 min before the beginning of intensive cell division. The ESR signal with g 2.2 (77 K) is caused by the spin-glass like structure. The "freezing" temperature of these spin-glasses was measured. PMID- 2553130 TI - The yeast ATP synthase subunit 4: structure and function. AB - The structure of ATP synthase subunit 4 was determined by using the oligonucleotide probe procedure. This subunit is the fourth polypeptide of the complex when classifying subunits in order of decreasing molecular mass. Its relative molecular mass is 25 kDa. The ATP4 gene was isolated and sequenced. The nucleotide sequence predicts that subunit 4 is probably derived from a precursor protein 244 amino acids long. Mature subunit 4 contains 209 amino acid residues and the predicted molecular mass is 23250 kDa. Subunit 4 shows homology with the b-subunit of Escherichia coli ATP synthase and the b-subunit of beef heart mitochondrial ATP synthase. By using homologous transformation, a mutant lacking wild subunit 4 was constructed. This mutant is devoid of oxidative phosphorylation and F1 is loosely bound to the membrane. Our data are in favor of a structural relationship between subunit 4 and the mitochondrially-translated subunit 6 during biogenesis of F0. PMID- 2553131 TI - Metabolic changes in undifferentiated and differentiated human colon adenocarcinoma cells studied by multinuclear magnetic resonance spectroscopy. AB - Aspects of energetic and intermediary metabolism were studied in a colon adenocarcinoma cell line (HT29) by multinuclear magnetic resonance spectroscopy. Experiments were carried out on the HT29-D4 clone, which was isolated by limit dilution techniques. This clone, usually undifferentiated (D4-UD), can be maintained in a differentiated state (D4-D) in a glucose-free medium. Metabolic data were obtained by NMR analysis of perchloric acid extracts from D4-UD and D4 D cells. Phosphorus-31 and proton NMR spectra showed the presence of a large amount of choline and phosphorylcholine in the differentiated state (400% and 200%, respectively, of the levels found in D4-UD cells). Other differences appeared in the content of phosphocreatine (absent in D4-D cells) and myoinositol (absent in D4-UD cells). Carbon-13 spectra were recorded from perchloric acid extracts of cells incubated with [1-13C]-labeled glucose or [2-13C]-labeled acetate. The data indicated that both types of cells metabolize glucose through the glycolytic pathway to give lactate, but only D4-D cells were able to store glucose as glycogen at a very high level. A mathematical analysis of fluxes through the tricarboxylic acid (TCA) cycle was developed on the basis of models derived from previous 14C tracer studies. The model was based on the steady-state labeling of glutamate carbons by the 13C isotope and gave the fraction of labeled acetyl-Coa entering the TCA cycle, and the activity y of anaplerotic reactions relative to the flux through the citrate synthetase reaction. The data indicated that y greater than 0.3 in all cases. Only 15% and 30% of labeled acetyl CoA entered the TCA cycle in D4-UD and D4-D cells, respectively, under labeled glucose incubation: these values were significantly different upon labeled acetate feeding, reaching 55% for D4-UD cells and 85% for D4-D cells. The main result of this study is that the process of differentiation of HT29 cells is correlated with a large increase in the activity of oxidative metabolism. PMID- 2553132 TI - [The effect of bilirubin and serum albumin on hydrolase activity in the synaptosomal membrane]. PMID- 2553133 TI - [Calcium transport in brain synaptosomes during depolarization. The role of potential-dependent channels and Na+/Ca2+ metabolism]. AB - The contribution of Ca2+ channels and Na+/Ca2+ exchange to Ca2+ uptake in rat brain synaptosomes upon long- (t greater than or equal to 30 s) and short-term (t less than 30 s) depolarization by high K+ was studied by measuring the 45Ca content and free Ca2+ concentration (from Quin-2 fluorescence). At 37 degrees C, the system responsible for the K+-stimulated uptake of 45Ca (t greater than or equal to 30 s) and the Na+/Ca+ exchanger are characterized by a similar concentration dependence of external Ca2+ (Ca0(2+] and K0+ as well as by an equal sensitivity to verapamil (Ki = approximately 20-40 microM) and La2+ (Ki = approximately 50 microM). These data and the results from predepolarization suggest that the 45Ca entry into synaptosomes at t greater than or equal to 30 s is due to the activation of Na+/Ca+ exchange caused by its electrogenic component, while the insignificant contribution of Ca2+ channels can be accounted for by their inactivation. At low temperatures (2-4 degrees C) which decelerate the inactivation, the initial phase of 45Ca uptake is fully provided for by Ca2+ channels, showing a lower (as compared to the exchanger) affinity for Ca0(2+) (K0.5 greater than 1 mM)m a greater sensitivity to La3+ (Ki = approximately 0.2 0.3 microM) and verapamil (Ki = approximately 2-3 microM); these channels are fully inactivated by predepolarization with K0+, ouabain and batrachotoxin. The Ca2+ channels can be related to T-type channels, since they are not blocked by nicardipine and niphedipine. PMID- 2553134 TI - [Protein composition of DNA-matrix complexes with "strong" and "weak" types of bonds, isolated from Ehrlich ascites carcinoma cells]. AB - A preparative procedure for obtaining tightly and loosely bound DNA-matrix complexes was developed. To this end, the DNA-matrix complexes were separated from DNA excess by restrictase digestion and solubilized in 0.1% sodium dodecyl sulfate + 8 M urea. To obtain tightly bound complexes, the lysate was transferred to a solution containing 2 M LiCl and 4 mM urea and gel filtered in the same medium. Tightly bound complexes were eluted in the same peak with DNA. Loosely bound complexes were obtained by gel filtration in 0.1% sodium dodecyl sulfate + 8 M urea after preliminary digestion of the original complexes with DNAase I which selectively destroys the tight bonds. Within the composition of loosely bound DNA-matrix complexes, the polypeptides with Mr of 180, 65-75, 58 and 47-50 kDa were identified, whereas tightly bound complexes were shown to contain polypeptides with molecular masses of 180, 65-75, 63, 61, 58 and 52-53 kDa. PMID- 2553135 TI - [Interconnection between activity and conformational mobility of alpha chymotrypsin in reverse micelle systems]. AB - A comparative study of the catalytic activity of alpha-chymotrypsin and the spin label rotation frequency in the alpha-chymotrypsin active center of reverse micellar systems solvated by H2O-organic mixtures was carried out. It was found that the decrease in the label rotation frequency resulting from the substitution of water in the micellar inner cavity by glycerol, 2.3-butanediol and dimethylsulfoxide (up to 95%) caused a marked increase (20-fold in the case of 2.3-butanediol) of the enzyme catalytic activity. The experimental results are quantitatively interpreted in terms of a simple kinetic scheme postulating the existence of the enzyme in two interconvertible forms differing in the conformational (intramolecular) mobility, i.e., the resting one predominantly existing in aqueous solution, and the tense one whose proportion rises with an increase in the concentration of the water-miscible organic solvent in the reverse micellar system. The value of kcat (2.4 s-1) for the tense form of the enzyme exceeded by more than 25 times the catalytic activity of chymotrypsin in aqueous solution (0.09 s-1) for the resting form. PMID- 2553136 TI - [The characteristics of [3H]muscimol binding to the synaptic membranes of the cerebral cortex in the progeny of rats subjected to deprivation of the paradoxical sleep phase during pregnancy]. AB - White rats were subjected to REM-sleep deprivation during 14-20 days of pregnancy. [3H]muscimol binding level of neocortex synaptosomal membranes was significantly higher in their offsprings as compared with the control ones. Possible ways of GABA-ergic system malformations are discussed. PMID- 2553137 TI - A study into the nature and organ source of digoxin-like immunoreactive substance(s) in the perinatal period. AB - Digoxin-like immunoreactive substance(s) (DLIS) was isolated from sera and autopsy-derived tissue obtained from premature and full-term neonates. The highest tissue level of DLIS was in the small bowel followed by the adrenal, gallbladder and liver. Of the fluids examined, meconium had the highest level of DLIS. Preparative high performance liquid chromatography fractionation of cord blood generated at least six different fractions which not only contained DLIS material but also inhibited canine kidney Na+/K+-ATPase activity. Recovery/inhibition studies indicated that 72% of the canine kidney Na+/K+-ATPase inhibition within one fraction could be accounted for on the basis of progesterone content of the fraction. PMID- 2553138 TI - Reduced levels of uridine diphosphate glucose pyrophosphorylase activity in fetal and neonatal human and guinea pig liver. AB - Uridine diphosphate glucose pyrophosphorylase (UDPGPP) is the first enzyme in the bilirubin conjugation pathway. A study aiming to screen for red blood cell UDPGPP deficiency in newborns with hyperbilirubinemia was carried out. No individuals with severe UDPGPP deficiency were found, however, levels of UDPGPP in premature and at term newborns were lower than in adults. These findings led to the study of UDPGPP in human fetal, neonatal and adult liver, using guinea pig tissues as a parallel control. UDPGPP activities in fetal and neonatal samples were also significantly lower than in adult ones in both species. Therefore, it is postulated that the reduced levels of UDPGPP in fetal and neonatal liver could be a factor which contributes to the pathogenesis of the physiologic jaundice in human newborns. PMID- 2553139 TI - Accumulation of diacylgylcerol in platelet phosphoinositide turnover in schizophrenia: a biological marker of good prognosis? AB - Phosphoinositide (PI) turnover was examined by measuring phosphatidic acid (PA) and diacylglycerol (DG) production following thrombin stimulation in platelet membranes from 20 schizophrenic patients, 10 patients with other mental disorders, and 9 normal controls. In 6 of 13 acute schizophrenic patients, DG was not transformed into phosphatidic acid (PA), but accumulated in the platelets instead. The abnormal findings persisted for at least 2 months, but then reversed over a long period. Three years later, the patients with abnormal PI turnover had a significantly better outcome than other acute schizophrenic patients. PMID- 2553140 TI - Preliminary evidence of benzodiazepine subsensitivity in panic disorder. PMID- 2553141 TI - Generation of reactive oxygen species, lipid peroxidation, and human sperm function. AB - Recent studies have demonstrated that human spermatozoa are capable of generating reactive oxygen species and that this activity is significantly accelerated in cases of defective sperm function. In view of the pivotal role played by lipid peroxidation in mediating free radical damage to cells, we have examined the relationships between reactive oxygen species production, lipid peroxidation, and the functional competence of human spermatozoa. Using malondialdehyde production in the presence of ferrous ion promoter as an index of lipid peroxidation, we have shown that lipid peroxidation is significantly accelerated in populations of defective spermatozoa exhibiting high levels of reactive oxygen species production or in normal cells stimulated to produce oxygen radicals by the ionophore, A23187. The functional consequences of lipid peroxidation included a dose-dependent reduction in the ability of human spermatozoa to exhibit sperm oocyte-fusion, which could be reversed by the inclusion of a chain-breaking antioxidant, alpha-tocopherol. Low levels of lipid peroxidation also had a slight enhancing effect on the generation of reactive oxygen species in response to ionophore, without influencing the steady-state activity. At higher levels of lipid peroxidation, both the basal level of reactive oxygen species production and the response to A23187 were significantly diminished. In contrast, lipid peroxidation had a highly significant, enhancing effect on the ability of human spermatozoa to bind to both homologous and heterologous zonae pellucidae via mechanisms that could again be reversed by alpha-tocopherol. These results are consistent with a causative role for lipid peroxidation in the etiology of defective sperm function and also suggest a possible physiological role for the reactive oxygen species generated by human spermatozoa in mediating sperm-zona interaction. PMID- 2553142 TI - Evidence that cyclic adenosine 3',5'-monophosphate-dependent protein kinase activation causes pig ovarian granulosa cell differentiation, including increases in two type II subclasses of this kinase. AB - Agents that elevated intracellular cyclic adenosine 3',5'-monophosphate (cAMP) caused a 3- to 10-fold increase in the luteinizing hormone (LH) receptor level and in progesterone biosynthesis in primary cultures of pig ovarian granulosa cells. Associated with these effects was a 2- to 4-fold increase in the total activity of the catalytic subunit of cAMP-dependent protein kinase in the tissue. From quantitation by [3H]cAMP binding and changes in the specific labeling with the photoaffinity analog [32P]-8-azido-cAMP, these agents were found to cause a concomitant 5- to 15-fold increase in two isoforms of the type II R-subunit (Mr = 54,000 and 56,000) of the protein kinase. Since the two intrasubunit cAMP binding sites of the protein kinase have been found to be positively cooperative, the addition of a combination of an analog selective for site 1 and an analog selective for site 2 causes synergistic increases in protein kinase activation in vitro and synergistic increases in intact cell responses if mediated by the cAMP dependent protein kinase. In the present study, the addition of such a combination of site 1- and site 2-selective analogs to granulosa cells caused a synergistic increase in LH receptor induction and progesterone production. For both responses, synergism did not occur when two analogs selective for the same site were combined. The results indicated that these responses are mediated by either of the two major isozyme types of cAMP-dependent protein kinase. PMID- 2553143 TI - Effects of polymyxin B on the sarcoplasmic reticulum membrane. AB - Polymyxin B, a cyclic peptide antibiotic, inhibits Ca2+-ATPase, p-nitrophenyl phosphatase and phosphorylase kinase activities associated with rabbit skeletal muscle sarcoplasmic reticulum membranes; 50% inhibition is induced by 100 microM, 130 microM and 550 microM of polymyxin respectively. The fluorescence intensity of fluorescein isothiocyanate-labeled Ca2+-ATPase, decreases in the presence of polymyxin (50% of the total decrease at 70 microM polymyxin). On the other hand, the polypeptide inhibits calmodulin-dependent endogenous phosphorylation of 60 kDa, 20 kDa and 14 kDa membrane proteins, while an increase of calmodulin dependent phosphorylation is observed in 132 kDa and 86 kDa proteins. PMID- 2553144 TI - Membrane viscosity of lymphocytes and influence of phytohemagglutinin. AB - The membrane viscosity of peripheral blood lymphocytes (PBLs) of equine, bovine and canine was measured by the use of time-resolved fluorescence depolarization technique with 1, 6-diphenyl-1,3,5-hexatriene (DPH). The viscosity values were 0.55, 0.59 and 0.50 poise for equine, bovine and canine PBLs, respectively. These values were compared with steady-state anisotropies and order parameters measured from electron spin resonance (ESR) of 5-doxyl stearic acid. Both values were increased with increase of viscosity. The fluid property of the membranes stimulated with phytohemagglutinin-P (PHA) was measured with steady-state fluorescence anisotropy and ESR. Little change of membrane fluidity was recognized with both methods during the stimulation with PHA. It appears that PHA activation process for these lymphocytes does not included large increase of the membrane fluidity which significantly accelerate the diffusion velocity of receptors in the plasma membrane. PMID- 2553145 TI - [Acid hydrolase activity and cyclic nucleotide contents in the rat heart during myocardial ischemia and postischemic reperfusion]. AB - The acid phosphatase and cathepsin D activities and cAMP and cGMP levels in isolated perfused rat heart were investigated during various periods of ischaemic myocardial injury and postischaemic reperfusion. The effect of phosphodiesterase inhibitor--caffeine was also studied. Free acid hydrolases activities and cyclic nucleotide content were increased under 40 and 60 min ischemia and 20 min postischaemic reperfusion. Addition of 50 microM caffeine to perfusion solution after 30 min of ischaemia resulted in increase of cAMP level, cAMP/cGMP ratio, lysosomal bound activities of acid hydrolase and decrease of free acid hydrolase activities. The obtained results suggested that defect in cAMP synthesis might be present in lysosomal membranes labilization in cardiomyocytes injured during ischaemic conditions. Addition of such agents, as caffeine, which increased heart cAMP level, may be effective in lysosomal membranes stabilization under reversible heart ischaemia and reperfusion. PMID- 2553146 TI - [Post-stress correlation of the functional activity of macrophages by tuftsin and its derivatives]. AB - The effect of 15 day programmed neurotization on the functional activity of the peritoneal macrophages has been studied. The NBT-test and the adhesion measurements were used. The experimental neurosis resulted in the decrease of the macrophage functional activity and in leukopenia. Tuftsin did not restore the stress induced depression of macrophage activity but led to additional rise of the adrenal glands weight and to pronounced granulocyte-monocytosis. Pentapeptide analog of tuftsin gave the additional inhibition of NBT-activity of the macrophage. Heptapeptide analog favoured the restoration of the macrophage activity after neurotization and stimulated lymphopoiesis. PMID- 2553147 TI - [Mechanism of the inhibition of proliferation of hapten-modified human and mouse leukemic cells by hapten-specific T-suppressor factor]. AB - T-suppressor-inducer and T-suppressor-effector hybridoma supernatants both regulating the DTH reaction intensity triggered by trinitrophenyl-sulphonic acid (TNPSA) or antigen-dependent proliferation of mice lymph node cells to trinitrophenylated bovine serum albumin, possess and ability to inhibit proliferation of TNPSA-modified human and mice leukaemic cells. This effect is specific, and is caused from antigen binding. This effect is specific, and is caused from antigen binding, idiotype positive T-suppressor factors and is based on a cyclic adenosine monophosphate increase of the concentration in target cells. PMID- 2553148 TI - [Role of sex dimorphism in the pharmacological action of etimizol]. AB - It has been shown that in female rats the level of ACTH, corticosterone in the blood, relative mass of adrenals, maintenance of T3, T4 in the serum and liver was significantly higher, but the activity of liver enzyme microsomes system was lower than in males; no sex differences were observed in myocardial creatine phosphokinase system. The influence of the etimizol on the female rats significantly speed up amidopyrine N-demethylation and biotransformation of hexobarbital. In males these systems react less on etimizol, but it reduces the speed of amidopyrine N-demethylation. PMID- 2553149 TI - [Retinoic acid modification of cell culture used for reproduction of enteropathogenic viruses]. AB - The 0.001-0.005% retinoic acid injection into the growth cultural medium of prime and continue cell cultures 12-24h before inoculation considerably raised the cell sensitivity to animal entero- and coronaviruses. The entero- and coronaviruses concentrations in cultural medium increased by 10(1.58) and 10(1.68)TCID 50/1.0 respectively. The optimized parameters of the cell culture processing for the enteropathogenic viruses reproduction improvement are proposed. PMID- 2553150 TI - [Sigma receptors of the loach embryo regulate ornithine decarboxylase activity]. AB - When loach embryos (Miagurnus fossilis) were incubated with opioid substance, sigma-receptors ligand SKF 10.047, activity of the key enzyme of polyamine synthesis - ornithine decarboxylase (ODC), increased. A correlation between teratogenic and ODC-stimulated action of SKF 10.047 was stage-specific and depend on opioid concentration. Possible mechanism of embryonic cell aggregation under SKF 10.047 action and polyamines role in that process is discussed. PMID- 2553151 TI - [Receptor of prolactin and its effect on binding of corticotropin in human and guinea pig adrenocortical cells]. AB - Specific binding of prolactin (PRL) by human and guinea pig isolated adrenocortical cells is saturable and reach equilibrium during 60 min. Characteristics of PRL binding by adrenocortical microsomes were determined. A single receptor species related to the high affinity and low binding capacity receptor class was revealed. The PRL-receptor affinities in adrenal cortex and liver are similar (approximately 10(9) M-1). Human adrenals have a higher affinity than that in guinea pigs. PRL increased corticotropin binding by adrenocortical cells of both species. Corticotropin binding rise may be due to the increase in the number of available receptors of low affinity and high capacity. PMID- 2553152 TI - [Biochemical and functional changes during immunization of rats with angiotensin II]. AB - Angiotensin-converting enzyme (ACE) activity in serum and some brain areas, level of angiotensin I in the blood and drinking behaviour during immunization of rats against conjugate of angiotensin II with bovine serum albumin (BSA) were studied. The results show that an increase in antibodies against angiotensin II was correlated with elevated ACE activity in serum. There was a distinct tendency towards elevated level of angiotensin I in the blood. After a 6 month's immunization ACE-activity was reduced twofold to threefold in midbrain and hypothalamus-thalamus. During immunization water-uptake was increased by 40-45%. PMID- 2553153 TI - [Inhibition by local anesthetics and barbiturates of the active transport of H+ in the synaptic vesicle membranes of the brain]. AB - The effects of local anesthetics and barbiturates on the ATP-dependent H+ transport in synaptic vesicle membranes from rat brain were studied using a fluorescent probe, acridine orange. Local anesthetics depressed the active H+ transport with the following order of potencies: tetracaine trimecaine lidocaine procaine. Respective IC50 values were 0.07, 0.28, 0.46 and 0.60 mM. The local anesthetics also disrupted the endogenous pH gradient seen in the absence of ATP. Barbiturates inhibited the active H+ transport showing IC50 values in the range of 2-5 mM except for benzobarbital and barbital characterized by IC50 values of 0.5 and 20 mM, respectively. The order of potencies was benzobarbital hexobarbital amobarbital pentobarbital phenobarbital barbital. The endogenous pH gradient was not affected by the barbiturates. The results show that local anesthetics disrupt the H+ transport by acting as permeable weak bases (uncouplers) whereas barbiturates are likely to block and anion channel which maintains electroneutrality of the H+ transport in the membrane of synaptic vesicles. PMID- 2553154 TI - [Dependence of the effects of tripeptide MIF and lithium chloride on the degree of supersensitivity of dopamine receptors of the brain]. AB - The desensitizing effects of MIF and lithium in respect to supersensitive striatal DA receptors in rats with unilateral lesion of the nigro-striatal pathway by 6-hydroxydopamine were revealed. Two groups of 6-hydroxydopamine denervated rats were selected by their qualitative responsiveness to apomorphine induced rotational behavior. It was found, that MIF and lithium (subchronic administration) did not modify behavioral supersensitivity in the highly sensitive group which showed two-peak rotational pattern in response to 0.05 mg/kg apomorphine, which was converted into a single-peak rotational pattern by haloperidol. On the contrary, administration of MIF and lithium for 21 days inhibited the apomorphine-induced rotations to 54 and 65% respectively in the less supersensitive group, which showed a single peak rotational pattern to apomorphine. Moreover, haloperidol showed the high antagonistic potency in these animals. These results suggest, that MIF and lithium might not exert desensitizing effects in the presence of high degree of supersensitivity of the striatal DA receptors, which is probably involved, for example, in the phenomenon of persistent tardive dyskinesia. PMID- 2553155 TI - [Effects of opioid receptor ligands on DNA synthesis and histamine contents in the gastric mucosa and blood of white rats]. AB - Using autoradiographic study with 3H-thymidine and spectrofluorometric method the authors studied the influence of opioid receptors' ligands on the DNA synthesis in the stomach epithelium histamine content and the blood. Leu-encephalin, B endorphin, dalargin, napoxone were administered intraperitoneally to male rats. The dose of injection was 0.1 ml per 100 g body weight. It was observed that B endorphin and dalargin 1.5-1.6 fold increased the quantity of DNA-synthesised nuclei in the epithelium of fundal stomach section. Leu-encephalin and dalargin increased the histamine concentration in the stomach, at the same time dalargin caused a rapid decrease of histamine concentration in the blood. Naloxone also decreased histamine concentration in the stomach. The obtained results are being discussed in connection with dalargin therapy of ulcerous diseases. PMID- 2553156 TI - [Effector modeling of the action of ligands of GABA-receptor complex. Functional interaction of the hypothetic alcohol receptor with other subunits of the complex]. AB - The types of the interaction of the pharmacological effects of ethanol and barbiturate antagonists--picrotoxin, bemegride and corasol--were determined. The effect of ethanol was determined as competitive--for the convulsant effects of bicuculline, and non-competitive--for the effects of thiosemicarbazide. The indices of the anticonvulsant effects of n-aliphatic alcohols were compared. It is suggested that n-aliphatic alcohols alter the functional status of the supramolecular GABA-receptor channel ensemble. The pharmacological properties and the elements of the structural similarity of picrotoxin and n-propanol (the presumptive ligand of the GABA-receptor channel ensemble) are discussed. PMID- 2553157 TI - [Cyclic analogs of ACTH fragments in self-stimulation and grooming behavior in rabbits]. AB - In present study new cyclic fragments of ACTH EHFRWGKPVG--NH2 and KHFRWG--NH2 were investigated in organization of self-stimulation and grooming behaviour in rabbits. Intracerebroventricular injections of EHFRWGPVG--NH2 in doses of 0.1-2.5 g evoked significant increases of self-stimulation and in doses of 4-5 g suppressed self-stimulation in rabbits. The effect of other fragments KHFRWG--NH2 on self-stimulation was not statistically significant. Both fragments induced excessive grooming behaviour in rabbits. The effects of these fragments persisted of 48-72 hours. PMID- 2553158 TI - Inhibition of chemotaxis Ng-monomethyl-L-arginine: a role for cyclic GMP. AB - The metabolism of L-arginine to nitric oxide (NO) has been shown to be important for the effector functions of many cell types, including polymorphonuclear (PMN) leukocytes. Its effect appears to be mediated at least in part by NO stimulation of soluble guanylate cyclase. We evaluated the role of this pathway in two PMN effector functions: cell movement and microbial killing, using the competitive inhibitor of L-arginine conversion to NO, NG-monomethyl-L-arginine (NMA). We also evaluated the effect of additional L-arginine and dibutyryl cyclic guanosine monophosphate (cGMP) on any NMA-associated changes. Human peripheral blood neutrophils were used and the cells were incubated with and without NMA. Chemotaxis was evaluated using a 48-well micro-Boyden chamber. Microbial killing was evaluated using S aureus strains D2C and 502A. These studies demonstrated that chemotaxis to formyl-methionyl-leucyl-phenylalanine was markedly inhibited in NMA-treated cells. This inhibition could be overcome if L-arginine or dibutyryl cGMP were added with the NMA. In contrast, microbial killing of S aureus was unaffected by NMA. These observations support the hypothesis that the L-arginine metabolism to NO and its effect on the cGMP level may be important for the dynamic changes required for neutrophil chemotaxis. PMID- 2553159 TI - Molecular heterogeneity in acute leukemia lineage switch. AB - Six cases of acute leukemia that underwent lineage switch from acute lymphocytic leukemia to acute myelogenous leukemia are reported. The mean age of the patients was 24 years, time to conversion was 36 months, and survival after conversion was only 3 months. Of the three cases which showed abnormal metaphases at both diagnosis and conversion, two (cases 2, 5) showed related cytogenetic abnormalities, and the third showed (case 3) independent chromosomal changes. Molecular analysis for immunoglobulin heavy chain and T-cell receptor beta chain genes showed that five of the six cases had rearrangement of at least one of these lymphoid associated genes at conversion to acute myelogenous leukemia. The single case (case 3) in which there were no lymphoid gene rearrangements at conversion was also the only case in which independent karyotypic abnormalities at diagnosis and conversion were demonstrated. Our findings suggest that lineage switch can represent either relapse of the original clone with heterogeneity at the molecular level or the emergence of a second new leukemic clone without molecular heterogeneity. PMID- 2553160 TI - Human myeloperoxidase gene expression in acute leukemia. AB - To evaluate the relationship between myeloperoxidase (MPO) gene expression and specific lineages of hematopoietic differentiation, we analyzed expression of the MPO gene in a variety of normal and leukemic human cells. As a first step, we synthesized several oligonucleotide probes and isolated cDNA clones for the MPO gene. MPO expression was detected in most acute myeloid leukemias, and the level of expression correlated well with cytochemical enzymatic activity. No MPO message was detected in pure lymphoid leukemias, whereas very low basal levels of MPO transcripts were found in normal bone marrows. In four cases of acute undifferentiated leukemias cytochemically negative for MPO, significant levels of MPO transcripts were detected, suggesting a myeloid origin for these cases. These results indicate that MPO gene expression may serve as an additional marker for subclassification of acute leukemias and may be used to identify leukemic cells arrested at an early stage of the myeloid differentiation pathway. PMID- 2553161 TI - Analysis of molecular events in leukemic cells arrested at an early stage of T cell differentiation. AB - We analyzed the rearrangement and expression of T-cell receptor (TCR) genes, including the recently identified TCR delta gene, in 21 patients with T-lineage leukemia/lymphoma. Among 8 patients with CD3-, CD4-, and CD8- (group I), 2 patients showed germline configuration of the TCR delta, gamma, beta, and alpha genes and 1 patient demonstrated only TCR delta gene rearrangement. All nine patients with CD3-, CD4+, and/or CD8+ (group II) showed concomitant rearrangements of the TCR delta, gamma, and beta genes. TCR alpha gene rearrangement was also observed in two patients. Three of four patients with CD3+ (group III) showed rearrangement of the TCR alpha gene with deletion of both alleles or of a single allele of the TCR delta gene. With Northern blot analysis, full-length transcripts of the TCR delta gene were detected in 3 of 15 examined patients. All were restricted to group I or group II. In contrast, full-length transcripts of TCR beta and alpha were observed mainly in samples from groups II and III. Based on these findings, rearrangement of the TCR delta gene may be the earliest event in T-cell differentiation, preceding rearrangements of the other TCR genes. PMID- 2553162 TI - Recombinant human granulocyte colony-stimulating factor as an activator of human granulocytes: potentiation of responses triggered by receptor-mediated agonists and stimulation of C3bi receptor expression and adherence. AB - Recombinant human granulocyte colony-stimulating factor (rhG-CSF) enhanced superoxide release and membrane depolarization in parallel in human granulocytes stimulated by the receptor-mediated agonists, N-formyl-methionyl-leucyl phenylalanine and wheat germ agglutinin, but not by the Ca2+ ionophore ionomycin and phorbol myristate acetate, which bypass the receptors to stimulate the cells. The optimal effect was obtained by pretreatment of cells with 25 to 50 ng/mL (1.3 to 2.6 nmol/L) rhG-CSF for 10 minutes at 37 degrees C. rhG-CSF produced by bacteria and mammalian cells had identical biological effects on a molar basis. rhG-CSF neither affected stimulus-induced increase in cytoplasmic free Ca2+ nor changed the number and affinity of N-formyl-methionyl-leucyl-phenylalanine receptors. The priming effect of rhG-CSF was temperature dependent and did not require new protein synthesis. rhG-CSF increased the expression of C3bi receptors on human granulocytes and enhanced granulocyte adherence to nylon fiber. The optimal effect was obtained by pretreatment of cells with 25 to 50 ng/mL rhG-CSF for 30 minutes at 37 degrees C. rhG-CSF had no effect on human monocytes. These findings demonstrate that rhG-CSF can selectively stimulate mature granulocyte functions. PMID- 2553163 TI - Recombinant human interleukin-3 and granulocyte-macrophage colony-stimulating factor show common biological effects and binding characteristics on human monocytes. AB - Two human hemopoietic growth factors involved in monocytopoiesis, interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) were studied for their ability to stimulate blood monocytes and to bind to the monocyte membrane. Both cytokines maintained monocyte/macrophage numbers during long-term culture and increased cell size as compared with controls. Effects on cell numbers were present at low cytokine concentrations (6 to 20 pmol/L), whereas enhanced 3H-thymidine incorporation was observed only at higher concentrations (greater than or equal to 60 pmol/L). Autoradiographic studies showed only 1% to 3% of stimulated monocytes with nuclear grains. These results suggest that the primary mechanism for IL-3 and GM-CSF-induced maintenance of monocyte/macrophage numbers in humans is through an effect on cell survival. Surface receptors for both IL-3 and GM-CSF were studied by using 125I-labeled recombinant human (rh) cytokines and performing Scatchard analyses. Both cytokines showed curvilinear Scatchard plots, and computer analyses favored a two-site binding model. High affinity binding data for 125I rhIL-3 (Kd 7.7 to 38.2 pmol/L; receptor number/cell 95 to 580) and for 125I rhGM-CSF (Kd 4.7 to 38.9 pmol/L; receptor number/cell 8 to 67) show similar binding affinities for the two cytokines but a lower receptor number/cell for 125I rhGM-CSF. Low-affinity binding characteristics for 125I rhIL-3 (Kd 513 to 939 pmol/L; receptor number/cell 179 to 5,274) and for 125I rhGM-CSF (Kd 576 to 1,120 pmol/L; receptor number/cell 130 to 657) show a similar pattern for the two cytokines. Specificity of 125I rhIL-3 and 125I rhGM-CSF binding to monocytes was established by the ability of the homologous cytokine to inhibit binding and the inability of a range of other cytokines to compete at 100-fold excess molar concentration. It is important, however, that binding of 125I rhIL-3 was partially inhibited by rhGM-CSF and that rhIL-3 partially inhibited binding of 125I rhGM-CSF to the monocyte membrane under conditions shown to prevent receptor internalization. The degree of inhibition varied between 25% and 80% in different experiments, and quantitative inhibition experiments showed that 1,000-fold excess concentrations of competitor failed to inhibit binding of the heterologous ligand completely. These results demonstrate that human IL-3 and GM-CSF have similar effects on growth and survival of human monocytes in vitro and suggest that these and other common biological effects may be mediated either through a common receptor or through distinct receptors associated on the monocyte membrane. PMID- 2553164 TI - A role for protein kinase C in the membrane fusion necessary for platelet granule secretion. AB - The addition of 1-oleoyl-2-acetylglycerol (OAG), or phorbol-12-myristate-13 acetate (PMA) to platelets induced the phosphorylation of a 47,000 dalton protein (47 Kd), fusion of granule membranes with membranes of the surface connected canalicular system, the formation of large vesicles and the secretion of serotonin. 1-(5-isoquinolinesulfonyl)-2-methyl-piperazine (H7), and sphingosine, inhibitors of protein kinase C, significantly inhibited the ultrastructural changes and the phosphorylation of 47 Kd. N-(2-guanidinoethyl)-5 isoquinolinesulfonamide (HA1004), structurally similar to H7, but a weaker inhibitor of protein kinase C, did not attenuate these responses to OAG or to PMA. H7, but not HA1004, also markedly inhibited secretion induced by the synergistic combination of OAG and the calcium ionophore A23187. Amiloride and 5 (N,N dimethyl)-amiloride, inhibitors of the Na+/H+ transporter, did not inhibit the ultrastructural response and the protein phosphorylation induced by OAG, or the secretion induced by the combination of A23187 and OAG. The results link the activation of protein kinase C by diglycerides to the labilization and fusion of granule membranes important for secretion. PMID- 2553165 TI - Increased density of ecto 5' nucleotidase antigen on leukemic T cells from patients with cutaneous T-cell lymphoma and adult T-cell leukemia/lymphoma. AB - Malignant CD4+ T cells in adult T-cell leukemia/lymphoma (ATL) and cutaneous T cell lymphoma (CTCL) express a number of cell surface molecules that are upregulated on normal T cells activated by foreign antigen. In this report we describe an interesting exception to the parallel phenotypic features of activated T cells and malignant CD4+ T cells. A monoclonal antibody (MoAb; termed 27.2) that was raised to HTLV-1+, CD4+25+ leukemic T cells stained weakly 25% of peripheral T cells, including approximately 50% of CD8+ T cells and 20% of CD4+ T cells. Flow cytometry analysis indicated that the surface density of the 27.2 antigen was unchanged or diminished when normal T cells were activated by antigen. However, 3/4 Sezary cases and 4/8 cases of ATL had relatively high densities of the 27.2 antigen. Immunoprecipitation and sodium dodecylsulfate polyacrylamide gel electrophoresis of the NP-40-solubilized membranes of surface iodinated ATL cells indicated that MoAb 27.2 reacted with a 75 Kd molecule. The size and distribution of the 27.2 antigen on T cell subsets suggested that it might be the enzyme ecto-5' nucleotidase (NT), a phosphatidylinositol-linked enzyme that catalyzes dephosphorylation of monophosphate nucleotides to their respective nucleosides. This was confirmed by demonstrating that lymphocyte ecto 5'NT activity was blocked partially and inhibited completely by preincubating cells with MoAb 27.2 for 1 hour at 4 degrees C and 24 hours at 37 degrees C, respectively. When used with a second MoAb (27.1) to a novel T cell activation antigen found on all CTCL and ATL leukemias examined, 27.2 was found to discriminate between normal and leukemic T cells in two patients with ATL. These studies suggest that ecto-5'NT has diagnostic value in T cell malignancies and may be aberrantly expressed in some cases of ATL and CTCL. PMID- 2553166 TI - Priming of human neutrophils with N-formyl-methionyl-leucyl-phenylalanine by a calcium-independent, pertussis toxin-insensitive pathway. AB - Resting neutrophils may be "primed" to augmented effector function, eg, superoxide (O2-) production in the respiratory burst, upon a second stimulation with a variety of soluble agonists including formylated methionyl-leucyl phenylalanine (FMLP) and phorbol myristate acetate (PMA). At priming concentrations of FMLP (5 x 10(-9) mol/L) that did not initiate O2- generation, two metabolic activities were noted: (1) approximately a threefold increase in the baseline intracellular calcium (Ca++i) level, that was not dependent on extracellular Ca++, and (2) a rapid rise in intracellular pH that was blocked by 5-(N,N-dimethyl) amiloride (DA), that had no effect on the Ca++i response to priming. Furthermore, there were no significant increases in inositol metabolites in cells primed and stimulated with FMLP compared with cells receiving the stimulating dose of FMLP alone and pretreatment with pertussis toxin (PT) (before the addition of the priming -5 x 10(-9) mol/L dose of FMLP), whereas abolishing the response to FMLP during the second stage of stimulation, had (1) no effect on FMLP-primed cells subsequently stimulated with PMA, and (2) only partially ablated the rise in Ca++i initiated with FMLP. That FMLP priming involved distinctive processes to those of the well characterized FMLP-coupled Ca++ dependent activation cascade was shown by the full priming effect attained in a Ca++-free buffer, which did not sustain an O2- response to a second-stage FMLP stimulation, but sustained a primed response to PMA. These data demonstrate that FMLP primes human neutrophils by a Ca++-independent and PT-insensitive pathway, offering a functional model for studying heterogeneous FMLP receptor-coupled reactions. PMID- 2553167 TI - Enhancement of erythrocyte superoxide dismutase activity: effects on cellular oxidant defense. AB - To delineate further the role of superoxide dismutase (SOD) in red blood cell (RBC) oxidant defense, normal human erythrocytes were osmotically lysed and resealed in the presence of varying concentrations of exogenous SOD. This resulted in a dose-dependent increase in SOD activity in the resealed erythrocytes while maintaining nearly normal RBC hemoglobin concentration (less than 10% decrease from the control value), cell volume, and cellular deformability. Surprisingly, a five- or ninefold increase in SOD activity yielded no additional protection against superoxide-generating drugs (phenazine methosulfate or menadione sodium bisulfite). No significant differences were observed between the control and SOD-loaded RBCs in O2-driven methemoglobin formation or generation of thiobarbituric acid-reactive substances. In contrast, RBCs with elevated SOD activity pretreated with sodium azide (to block catalase activity) or 1-chloro-2,4-dinitrobenzene (to deplete reduced glutathione, GSH) showed significantly enhanced methemoglobin generation in response to superoxide generating drugs. No differential response was noted between the control, control resealed, and SOD-loaded RBCs to oxidants other than superoxide. Based on our results and other data, we conclude that elevated SOD activity may imbalance cellular oxidant defense, resulting in enhanced oxidation due to the accelerated generation of H2O2, the product of O2- dismutation. This effect is significantly exacerbated under conditions in which H2O2 catabolism is altered. PMID- 2553168 TI - Cadmium inhibits brain calmodulin: in vitro and in vivo studies. PMID- 2553169 TI - Therapeutic efficacy of dimercaptosuccinic acid and thiamine/ascorbic acid on lead intoxication in rats. PMID- 2553170 TI - Isolation of glycoprotein D from herpes simplex virus type 1 by gel filtration high performance liquid chromatography. AB - Rabbit kidney (RK-13) and human jejunum and ileum (I-407) cells infected with herpes simplex virus type 1, strain F, were radiolabelled with [14C]glucosamine or [35S]methionine for 24 h. The cells were extracted with 1% Triton X-100 and the extracts were separated by gel filtration high performance liquid chromatography. Monoclonal antibody immunoprecipitation of the fractions collected from the column revealed a monomeric glycoprotein D (gD) of 52 - 56,000 molecular weight from RK-13 cells and two monomeric forms of gD, 54,000 and 58,000 molecular weight, from I-407 cells. Densitometry scanning of the autoradiograms from SDS-PAGE showed gD from the RK-13 host cells to be 98.7% pure with the [35S]methionine label and 97.0% pure with the [14C]glucosamine. On the other hand, gD from the I-407 host cells was only 78.6% with the [35S]methionine label and 96% pure with the [14C]glucosamine. This method could provide a means for the isolation of native gD for structural and immunological studies. PMID- 2553171 TI - Rapid structural analysis of the in vitro and in vivo metabolism of SK & F 95448 by the combined use of thermospray liquid chromatography/mass spectrometry and liquid chromatography/tandem mass spectrometry. AB - The thermospray liquid chromatography/mass spectrometry (LC/MS) interface has had a major impact on the direct analysis of the metabolic fate of xenobiotics in complex biological media. This paper outlines the rapidity and power of the LC/MS approach, and shows how detailed structural information can be obtained without recourse to individual compound isolation. This provides a great saving in time and effort. The additional specificity of liquid chromatography/tandem mass spectrometry is highlighted in identifying the sites of metabolic transformation. The ability to handle biological samples with little or no clean-up using wide high-performance liquid chromatographic gradients is a key feature of the success of this methodology. PMID- 2553172 TI - Targeting of proteins for degradation. PMID- 2553173 TI - Mumps, measles, and rubella vaccination and encephalitis. PMID- 2553174 TI - Subcellular fractionation of epithelial cells from toad urinary bladder. 2. Isopycnic centrifugation and effect of density perturbants. AB - Cytoplasmic granules obtained from toad urinary bladder epithelial cells were brought to buoyancy in a linear sucrose gradient. The gradient was loaded either with untreated cytoplasmic granules, or with granules treated with Na pyrophosphate (PPi), with digitonin, or with PPi and digitonin in succession. The following enzymes were assayed in the gradient subfractions: oligomycin insensitive Mg++-ATPase, alkaline phosphodiesterase I, alkaline phosphatase, acid N-acetyl-beta-glucosaminidase, cytochrome oxidase, nucleoside diphosphatase (substrate, ADP), aminopeptidase (substrate, leucyl-beta-naphthylamide), and mannosyltransferase (acceptor, dolichylphosphate). Comparison of the density distributions of enzymes in untreated and treated preparations led to the characterization of 4 distinct subcellular entities. In agreement with the properties of mitochondria from other cell types, cytochrome oxidase buoys at 1.18 within a narrow density range and its behavior is not significantly altered by PPi or digitonin. Under all conditions, acid N-acetyl-beta-glucosaminidase is recovered over a broad density range in the lower part of the gradient and appears as a qualified lysosomal marker. Mg++-ATPase, alkaline phosphodiesterase I, and alkaline phosphatase belong to a group with the distinguishing features of a low equilibrium density in native cytoplasmic granules and a marked shift (+0.03 density units) after digitonin treatment. Such properties are typical of the plasma membranes. Part of the aminopeptidase activity probably also belongs to plasma membrane-derived elements. Minor differences between alkaline phosphatase and the other 2 members of that group make it possible that their distribution domains in the membrane do not overlap or coincide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553175 TI - (Na,K)-pump: cellular role and regulation in nonexcitable cells. AB - The (Na,K)-pump develops and maintains ionic gradients that are of fundamental importance for proper function of most animal cells. These gradients are utilized in the form of ionic leak pathways by a number of special and general cell processes (e.g., nerve conduction, nutrient transport, pH regulation). As the sodium gradient in particular energizes many vital cell processes, alterations in cell activity will often be manifest as changes in sodium entry. The (Na,K)-pump rate varies accordingly, in order to maintain balance between Na entry and exit thereby maintaining the potential energy of the cell. Acute changes in sodium influx are balanced by increases in activity of existing pump units, with only a small change in intracellular sodium concentration. This is possible because intracellular is normally poised on the steep limb of the concentration versus activity curve for the (Na,K)-pump, at a point well below maximal activity, allowing large increases in (Na,K)-pump rate with only small changes in sodium concentration. If the increase in sodium influx is prolonged, it appears that the cell responds by synthesizing new pumps, allowing intracellular sodium concentration to return to its original values. Though increases in (Na,K)-pump activity must be accompanied by increases in potassium leak rates, in the experiments we have presented, there does not appear to be direct functional coupling between (Na,K)-pump and the K leak pathways. In these situations the matching of active influx and passive efflux of K short-term appears to occur by mechanisms not directly related to (Na,K)-pump activation. PMID- 2553177 TI - Synaptosomal apyrase in the hypothalamus of adult rats. AB - 1. The synaptosomal fraction isolated from hypothalamus of adult rats on a sucrose density gradient hydrolyzes the labile phosphates from ATP and ADP, thereby satisfying the general definition of apyrase activity. 2. The parallel behavior of ATPase and ADPase activities under different reaction conditions suggests the presence of a "true" apyrase enzyme. The optimum conditions for the reaction are the same for both nucleotides: pH 8.0, 0.6 mM nucleotide and 1.5 mM cation. At temperatures between 10 and 40 degrees C, both activities increase with no change in the ATP/ADP hydrolysis ratio. Thermal inactivation or inhibition of the enzyme activity by iodoacetamide, p-hydroxymercuribenzoate or 2 mercaptoethanol affected the hydrolysis of both substrates in a similar manner. 3. Adenylate kinase and pyrophosphatase activities were not detected in the preparation. 4. The enzyme is located on the outer surface of the synaptosomal membrane: intact and lysed synaptosomes have similar activity and the supernatant obtained by centrifugation of intact synaptosomal preparations does not hydrolyze ATP or ADP. PMID- 2553176 TI - Water flow in the toad urinary bladder in response to vasopressin: role of potassium. AB - In agreement with previous reports, we found that absence of K+ from the serosal bath of the toad urinary bladder substantially impairs vasopressin and cAMP stimulated water flow. The decreased response to vasopressin appears unrelated to prostaglandin production since inhibition of endogenous prostaglandins by pretreatment with naproxen 10(-5) M failed to prevent the effect seen with K+ free Ringer's. The resistance to vasopressin does not appear to be directly related to epithelial K+ concentrations, in that maneuvers leading to decreased intracellular K+ failed to produce a similar effect. A more likely explanation appears to be that K+-free Ringer's induces an increased cytosolic Ca++ which, in turn, decreases the hydrosmotic effects of vasopressin. Several lines of evidence argue in favor of such an explanation: (a) Increased cytosolic Ca++ had been found in other tissues with low extracellular K+; (b) The resistance to vasopressin decreases with decreased serosal Ca++; (c) The effects of K+-free Ringer's are not additive in situations believed to have increased epithelial Ca++, i.e. replacement of serosal Na+ with choline; (d) The effects of K+-free serosal bathing medium could be both prevented and/or reversed if already established by increasing serosal bath, and presumably intracellular, pH, which is believed to decrease intracellular Ca++. PMID- 2553178 TI - The effect of short-wavelength ultraviolet light on antigens, lectin receptors and the ultrastructure of Crithidia fasciculata. AB - Crithidia fasciculata is an important trypanosomatid parasite commonly affecting insects and is used extensively as a model for the study of the biochemistry, ultrastructure and organization of the kDNA network of trypanosomatids. The present study describes the evolution of UV-induced morphological changes detectable by transmission electron microscopy in Crithidia fasciculata. Although only rare and minor changes in kinetoplast DNA were demonstrable 7 h after UV irradiation, alterations of this organelle were present in almost all flagellates observed 24 h and 48 h after irradiation. Other cell structures were apparently undamaged. Ultrastructural changes in kDNA did not correspond to changes in antigenicity of protein bands in western blotting against serum from Chagas' disease patients or in the presence of 3 different lectin receptors on the surface of the parasite. PMID- 2553179 TI - Experimental vaccination of rabbits with non-pathogenic strains of Aujeszky's disease virus. AB - 1. The Bartha K and NIA-4 strains of Aujeszky's disease virus (ADV) were non pathogenic for rabbits vaccinated once or twice by nasal instillation or intramuscular injection. Neutralizing antibodies were detected in 68% of the rabbits two weeks after primary vaccination and in all rabbits at challenge. 2. Challenge doses of virulent ADV greater than 10(5.0) median tissue culture infective doses (TCID50) resulted in the death of most vaccinated and all unvaccinated rabbits with typical signs of Aujeszky's disease within 4 days. ADV was recovered from brain and lung suspensions of vaccinated and unvaccinated rabbits who had died as a result of the challenge. 3. When the challenge dose was reduced to approximately 10(3.0) TCID50, rabbits vaccinated twice survived while all unvaccinated controls died within 3 days. PMID- 2553180 TI - [Clinical and biological aspects of intensive chemotherapy with autologous bone marrow grafts in small-cell bronchial cancer]. AB - Small cell lung cancer (SCLC) is a frequent and aggressive tumor characterized by its major sensitivity to chemotherapy since 80% of the patients respond dramatically to the treatment. Nevertheless, the majority of them eventually die from their cancer. We have addressed the effect of late intensification with autologous bone marrow transplantation on SCLC through a randomized clinical trial. This study included 101 patients, 45 of whom could be randomized to either conventional post-induction therapy or late intensification. Of the 32 patients with limited disease, patients who did not receive intensification relapsed on a median of 10 weeks compared with 35 weeks for patients who received intensification (p = 0.001). An important observation was that there was no patient disease free at 1 year in the group that did not receive intensification whereas there were patients who were free of disease up to 271 weeks in the group receiving late intensification. To improve upon these results we have developed a panel of anti-lung cancer monoclonal antibodies that we use for the detection of occult cancerous disease in the bone marrow and for the in vitro elimination of clonogenic tumor cells contaminating the marrow graft. In addition we are monitoring the immune defect induced by ABMT and its compensation by an in vitro recombinant human IL-2 perfusion. Finally we have devised a new induction and intensification chemotherapy regimens with different drugs in order to prevent multidrug resistance. These new tools are currently under investigation in the clinical setting. PMID- 2553181 TI - GABA receptors: are cellular differences reflected in function? AB - The putative involvement of GABAA and GABAB receptors in various behavioral and physiological effects is summarized in Table III. A division of function among the two types of GABA receptors appears to exist. GABAA receptors mediate feeding, cardiovascular regulation, anxiolytic effects, and anticonvulsive activity. GABAB receptors, on the other hand, are involved in analgesia, cardiovascular regulation, and depression. Although there is some overlap and shared functions among the receptor types, it is evident that GABAA and GABAB receptors have different behavioral and physiological profiles. Feeding, anticonvulsive activity and anxiety, for example, primarily involve GABAA receptors. Analgesia and depression, on the other hand, are GABAB effects. In those cases where GABAA and GABAB receptors mediate similar functions (e.g. cardiovascular regulation), they do so by affecting different transmitter systems and cellular mechanisms. It is proposed, therefore, that GABAA and GABAB receptors differ not only at the cellular level, but that they also have different functions in the mammalian central nervous system. The association of different subtypes of a receptor with different functions and mechanisms of action is not unique to the GABA system. D1 and D2 receptors in the dopamine system, for example, also exhibit some separation of function as do the mu, delta and kappa types of opiate receptors. Different subtypes of neurotransmitter receptors, therefore, appear to be a general organizing principle used by the brain to transduce chemical signals into different functional responses. A better understanding of the exact processes through which cellular signals are transformed into functional responses is a goal of future research. PMID- 2553182 TI - Calcium, neuronal hyperexcitability and ischemic injury. AB - Due to tight regulatory controls, a 10,000-fold concentration gradient exists between intracellular and extracellular free Ca2+ concentrations. With appropriate stimulus Ca2+ will rapidly flow into neurons through various types of membrane channels including voltage-dependent and receptor-operated channels. Intracellular Ca2+ concentrations are then quickly restored primarily through Ca2+-ATPase, Na+/Ca2+ exchange, and endoplasmic reticulum sequestration. It is well-known that Ca2+ is essential for neurotransmitter release. More recent investigations indicate that Ca2+ influx is essential for neuronal excitability independent from synaptic function. In fact, abnormal Ca2+ metabolism may play a dominant role in both the initiation and propagation of seizure discharge. Accordingly, Ca2+ channel blockers may represent a new therapeutic modality to treat epilepsy. Analyzed in this article are the major mechanisms by which neurons control Ca2+ fluxes and the evidence supporting the role of Ca2+ in seizure phenomena. Thereafter, an integrative theory for the role of calcium in neuronal hyperexcitability and ischemic cell death is constructed. PMID- 2553183 TI - Restricted usefulness of tetraethylammonium and 4-aminopyridine for the characterization of receptor-operated K+-channels. AB - 1. Recently, we suggested that the D2-dopamine receptor involved in the inhibition of evoked [3H]-acetylcholine release from rat striatum is coupled to K+-channels. 2. In the present study, an attempt was made to elucidate further the role of these K+-channels, using the K+-channel blocking agents tetraethylammonium and 4-aminopyridine. With a superfusion method, the effects of both drugs on the D2-dopamine receptor-mediated inhibition of the electrically evoked release of [3H]-acetylcholine from rat striatal tissue slices was investigated. 3. Both tetraethylammonium (30 mM) and 4-aminopyridine (0.1 mM) significantly stimulated the electrically evoked release of [3H]-acetylcholine and completely abolished the effect of the selective D2-receptor agonist LY 171555 (1 microM) on evoked acetylcholine release. In addition, tetraethylammonium (0.03-30 mM) and 4-aminopyridine (0.003-1 mM) strongly increased the basal (non-evoked) release of radioactivity in a concentration dependent manner. The results suggest that the effect of the drugs on the basal release of radioactivity and on the electrically evoked release of acetylcholine cannot exclusively be explained by their action on K+-channels. 4. Furthermore, with the use of a receptor binding assay, data were obtained on the affinity of tetraethylammonium and 4-aminopyridine for D2-receptors and various other neurotransmitter recognition sites. At concentrations in which both drugs are known to block K+-channels, they were found to inhibit the specific binding of selective radioligands to their respective recognition sites. 5. It is concluded that due to their 'side-effects', both tetraethylammonium and 4-aminopyridine are of only limited value in the investigation of the alleged interaction between neurotransmitter receptors and K+-channels. PMID- 2553184 TI - Intracellular platelet-activating factor regulates eicosanoid generation in guinea-pig resident peritoneal macrophages. AB - 1. The role of intracellular platelet-activating factor (Paf) in arachidonic acid (AA) mobilization from guinea-pig peritoneal macrophages has been investigated by use of the potent and selective Paf receptor antagonists, WEB 2086 and CV 6209. 2. Adherent macrophages contained cell-associated Paf which was increased by exposure to formyl-methionyl-leucyl-phenylalanine (fMLP), endotoxin and the ionophore, A23187. However, only endotoxin and A23187 caused release of detectable amounts of Paf into the extracellular medium. 3. Exogenous Paf and each of the above stimuli mobilized previously incorporated [14C]-AA and increased the generation of prostacyclin (PGI2) in resident macrophages. 4. WEB 2086 (10-100 microM) and CV 6209 (0.1-10 microM) reduced both basal and stimulated PGI2 generation and WEB 2086 inhibited the mobilization of [14C]-AA. In addition, WEB 2086 (10 microM) inhibited fMLP-and Paf-induced superoxide anion generation. Responses to A23187 were not inhibited by either antagonist. 5. Activation of macrophages by fMLP caused a short burst of intracellular Paf generation but none was detected in the supernatants. The time-course of PGI2 synthesis followed closely that of Paf. 6. These data suggest that intracellular Paf generation is important for subsequent AA mobilization and may have a wider role in signal transduction processes. PMID- 2553185 TI - Different time courses of the blockade of sodium current by lignocaine and SUN 1165 in single myocytes isolated from guinea-pig atrium. AB - 1. The time course of the blockade of sodium currents (INa) by the antiarrhythmic agents, lignocaine and SUN 1165, was studied in single myocytes isolated enzymatically from guinea-pig atrium, by a new concentration-jump termed as a 'concentration-clamp' technique. This technique combines an intracellular perfusion and a rapid exchange of external solution surrounding the voltage clamped myocyte within 2 ms. 2. Lignocaine (3.7 x 10(-5) M to 3.7 x 10(-4) M) inhibited the peak amplitude of INa in a concentration-dependent fashion. It took 2 to 5s to reach apparent steady-state inhibition at the concentrations used. Complete recovery from the inhibition also took 2 to 5s after washing out the agent. In contrast, the inhibitory effect of SUN 1165 (1 x 10(-5) M) on the peak INa gradually progressed and reached a steady-state level about 2 min after the start of drug-application. The recovery required more than 10 min after washing out of the agent. 3. In cardiomyocytes treated with scorpion toxin (5 micrograms ml-1), the inactivation of INa was greatly inhibited, resulting in the relatively persistent Na inward current (persistent INa) during the depolarizing pulse. Lignocaine (1.1 x 10(-4) M) applied during the depolarizing pulse, reduced in a single-exponential fashion the amplitude of the persistent INa in milliseconds. On the other hand, lignocaine applied several tens of milliseconds before the depolarizing pulse induced only a small reduction of the peak amplitude of the first persistent INa. When SUN 1165 (1 x 10- M) was applied during the depolarizing pulse, there was only a small instantaneous reduction of the amplitude of the persistent I'N, although it did inhibit time-dependently, the peak IN.. Both agents accelerated the decay phase of the persistent IN in a time dependent manner. 4. These results suggest that lignocaine and SUN 1165 may preferentially interact with the openstate of the sodium channel rather than with the rested one, although SUN 1165 does so much more slowly. PMID- 2553186 TI - Electrophysiological effects of OPC-88117, a new antiarrhythmic agent on papillary muscles and single ventricular myocytes isolated from guinea-pig hearts. AB - 1. The effects of OPC-88117, a new antiarrhythmic agent, on transmembrane action potentials were examined in right ventricular papillary muscles and in single ventricular myocytes isolated from guinea-pig hearts. 2. In papillary muscles, OPC-88117 above 3 x 10(-6) M caused a dose-dependent prolongation of action potential duration (APD). 3. OPC-88117 above 3 x 10(-5) M caused a significant decrease in the maximum upstroke velocity (Vmax) of the action potential without affecting the resting membrane potential. The inhibition of Vmax was enhanced at higher stimulation frequencies. 4. In the presence of OPC-88117, trains of stimuli at rates greater than or equal to 1.0 Hz led to a use-dependent inhibition of Vmax with rapid onset. The time constant for the recovery of Vmax from the use-dependent block was 456 ms. 5. The curves relating membrane potential and Vmax were shifted by OPC-88117 to the direction of more negative potentials (9 mV at 10(-4) M). 6. In single ventricular myocytes treated with OPC 88117 (1-3 x 10(-4) M), the Vmax of test action potentials preceded by conditioning clamp pulses to 0 mV was decreased progressively as the clamp pulse duration was prolonged. 7. These findings suggest that the primary electrophysiological effect of OPC-88117 on the cardiac muscle cell is prolongation of APD (Class III action) and that at high concentrations, it may also possess a lignocaine-like sodium channel inhibitory effect (Class I action). PMID- 2553187 TI - Effects of OPC-88117, a new antiarrhythmic agent, on the electrophysiological properties of rabbit isolated hearts. AB - 1. Effects of a new antiarrhythmic agent, OPC-88117, on the conduction properties and excitability of Langendorff-perfused rabbit hearts were compared with those of lignocaine. 2. OPC-88117 above 10(-5) M caused a significant prolongation of atrio-His bundle conduction time (A-H interval) as well as His bundle-ventricular conduction time (H-V interval). Lignocaine above 10(-5) M caused a similar prolongation of H-V interval with a minimum change of A-H interval. The conduction time from His bundle stimulus to the left ventricle (St-LV) at a low frequency (1 Hz) was also prolonged by these drugs around 10(-4) M. 3. OPC-88117 above 3 x 10(-6) M prolonged the effective and functional refractory periods of His bundle (ERPHB, FRPHB) as well as the effective refractory period of left ventricular muscle (ERPVM) in a dose-dependent manner. ERPHB, FRPHB and ERPVM were shortened by low concentrations of lignocaine (3 x 10(-6)-10(-5) M), but were prolonged by high concentrations (3 x 10(-5)-10(-4) M). 4. Lignocaine (3 x 10(-6)-10(-4) M) induced an upward displacement of the His bundle refractory curve, indicating a greater intraventricular conduction delay for premature excitation. In experiments with OPC-88117, such an upward displacement was observed only at 10(-4) M. 5. These results suggest that the primary electrophysiological effect of OPC-88117 is a lengthening of ventricular refractoriness, and that at high concentrations it may also exert lignocaine-like inhibition of ventricular conduction. PMID- 2553188 TI - Selectivity of bradykinin analogues for receptors mediating contraction and relaxation of the rat duodenum. AB - 1. Bradykinin produces a biphasic response in the rat duodenum that consists of a relaxation (pD2 = 8.44) followed by a contraction (pD2 = 6.91). 2. The B1 agonist des-Arg9-bradykinin produced a contraction (pD2 = 7.16) but no relaxation. Des Arg9-[Leu8]-bradykinin, which is a B1 antagonist in other systems produced contraction (pD2 = 7.65) in the rat duodenum. 3. Four bradykinin analogues that are preferential B2 agonists in other tissues had a biphasic effect with pD2 values in the range 7.22-8.68 for relaxation and 6.26-6.91 for contraction. 4. [Thi5,8,D-Phe7]-bradykinin, which is a B2 antagonist in most other systems produced relaxation in the rat duodenum, with a pD2 of 7.49. 5. It is concluded that the contractile component of the response to bradykinin in rat duodenum may be mediated by a subtype of the B1 receptor and the relaxant component by a receptor of the B2 subtype. PMID- 2553189 TI - The pharmacological evaluation of LY 170680, a novel leukotriene D4 and E4 antagonist in the guinea-pig. AB - 1. This paper describes the evaluation of LY170680 (5-less than 3-[2(R) (carboxyethylthio)-(S)-hydroxypentadeca 3(E)5(Z)-dienyl]phenyl greater than 1H tetrazole, as an antagonist of the cysteinyl leukotrienes C4, D4, E4, in a variety of in vitro and in vivo models. 2. In vitro, LY170680 was a potent, selective, and competitive antagonist of LTD4, and LTE4. It produced a concentration-dependent rightward displacement of the concentration-response curves elicited by either LTD4 or LTE4 on both the guinea-pig ileal and tracheal preparation. The pA2 values for LY170680 were estimated to be 8.1 +/- 0.2 (n = 8) and 8.1 +/- 0.1 (n = 6) on trachea, and 8.7 +/- 0.1 (n = 12) and 9.0 +/- 0.3 (n = 6) on ileum for both LTD4 and LTE4 respectively. The slopes of the Schild plots in these studies were all close to unity. 3. LY170680 was shown to be a modest antagonist of the LTC4-induced responses on guinea-pig ileum (pA2 7.0 +/- 0.2 n = 5), but had no discernable effects against contractions induced by histamine, prostaglandin E2 (PGE2), PGF2 alpha or acetylcholine. The compound also reduced the LTC4-induced responses on trachea, but in a non competitive manner. 4. Intravenous LY170680 reduced in a dose-dependent manner (ED50 3.8 mg kg-1, 60 min pretreatment) the fall in compliance in the anaesthetized guinea-pig, and the rise in total pulmonary resistance (TPR) (ED50 2.0 mg kg-1, 30 min pretreatment) in the artificially ventilated guinea-pig, produced by intraveous LTD4. 5. LY170680 (5mgkg-1, i.v.) was also effective in reducing the increase in TPR to intravenous antigen in sensitized animals pretreated with mepyramine, indomethacin and propranolol. 6. The compound was only moderately effective (ED5o 40mgkg-1 p.o.) in preventing the rise in TPR induced by intravenous LTD4, when given orally. 7. Inhaled LY170680 was particularly effective in preventing the increase in TPR produced by an exposure to aerosolised LTD4. An estimated 1-2 pg of LY170680 delivered to the airways by nebuliser 1 hour beforehand, produced a 6 fold lateral displacement to the right of the dose-response curve to LTD4. 8. Studies in conscious animals indicated that inhaled LY170680 produced a dose dependent reduction in the increased volume of gas, trapped in the lung following exposure to aerosolised LTD4. Duration studies also indicated that an estimated inhaled dose of 10 g LY170680 significantly reduced the LTD4-induced' increase in trapped lung gas volume for at least 4 h. Inhaled LY170680 also reduced LTC4 induced increase in gas trapping in a manner similar to LTD4. However, histamine induced gas trapping was unaffected. PMID- 2553190 TI - Inotropic response to DPI 201-106 in the failing human heart. AB - 1. The present study was designed to characterize the positive inotropic response to DPI 201-106 in isolated papillary muscle strips obtained from heart failure patients undergoing surgery. 2. The positive inotropic responses to isoprenaline and milrinone and cardiac beta-adrenoceptor density were also determined. 3. DPI 201-106 increased the force of contraction in papillary muscle strips from patients with moderate (NYHA II-III) and severe (NYHA IV) heart failure, in a concentration-dependent manner. This positive inotropic effect was more pronounced in tissues from NYHA IV patients. Furthermore, these responses were greater than those produced by milrinone or isoprenaline. The positive inotropic effects of isoprenaline and milrinone were reduced in NYHA IV compared to NYHA II III. Consistently, there was also a smaller density of beta-adrenoceptors in myocardium from NYHA IV than in NYHA II-III. The positive inotropic effect of Ca2+ was similar in tissues from both groups of patients. 4. The positive inotropic effect of DPI 201-106 was not antagonized by adenosine or carbachol, whereas both compounds reduced the positive inotropic effect of isoprenaline. 5. DPI 201-106 did not increase the Ca2+ -sensitivity of chemically skinned ventricular fibres, whereas a significant increase of the Ca2+ -sensitivity was obtained with trifluoperazine. 6. It is concluded that DPI 201-106 produces significant positive inotropic effects in tissue excised from the failing human heart. The lack of inhibition by adenosine and carbachol might contribute to its greater effectiveness in NYHA IV than NYHA II-III and indicates that its mechanism of action is cyclic AMP-independent. A sensitization of the contractile proteins to Ca2+ does not appear to be important for the positive inotropic action of DPI 201-106 in the failing human heart. PMID- 2553192 TI - 5-HT1D binding sites in porcine brain can be sub-divided by GR43175. AB - We have examined the binding of 5-carboxamidotryptamine (5-CT) and GR43175 (3-(2 dimethylamino)ethyl-N-methyl-1H-indole-5-methane sulphonamide) to 5-HT1D sites labelled with [3H]-5-hydroxytryptamine [( 3H]-5-HT) in neonatal porcine caudate membranes. In competition studies, 5-CT produced shallow inhibition curves (Ki 138 nM, slope 0.31), indicating binding site heterogeneity, while GR43175 interacted with a single population of binding sites (Ki 251 nM, slope 0.98), producing a maximum of only 52% inhibition of [3H]-5-HT binding compared to 100% for 5-HT or 5-CT. In the presence of excess GR43175 (10 microM), 5-CT produced a monophasic inhibition curve with a Ki value of 800 nM for the remaining sites (slope 0.89). These preliminary data suggest that under the conditions employed, GR43175, and to a lesser extent 5-CT, may discriminate between two sub populations of 5-HT1D binding sites in porcine brain. PMID- 2553191 TI - The modulation by chlormethiazole of the GABAA-receptor complex in rat brain. AB - 1. The interactions of chlormethiazole with gamma-aminobutyric acid (GABA) synthesis and release, and with ligand binding to sites associated with the GABAA receptor complex and the GABAB-receptor have been studied in the rat. The GABAA receptor was studied using [3H]-muscimol, [3H]-flunitrazepam was used to label the benzodiazepine modulatory site, and [35S]-butyl-bicyclophosphorothionate ([35S]-TBPS) to label the chloride channel. 2. Chlormethiazole had no effect on GABA synthesis in the cortex, hippocampus and striatum or on GABA release from cortical slices in vitro. Chlormethiazole did not displace [3H]-baclofen binding to the GABAB-receptor. 3. Chlormethiazole (IC50 = 140 microM) and pentobarbitone (IC50 = 95 microM) both inhibited [35S]-TBPS binding by increasing the rate of [35S]-TBPS dissociation. In addition, chlormethiazole caused an apparent decrease in the affinity of [35S]-TBPS binding. 4. Chlormethiazole enhanced the binding of [3H]-muscimol but had no effect on [3H]-flunitrazepam binding. In contrast, the sedative barbiturate pentobarbitone enhanced both [3H]-muscimol and [3H] flunitrazepam binding. 5. It is concluded that the sedative and anticonvulsant effects of chlormethiazole are probably mediated through an action at the GABAA receptor. However, chlormethiazole does not interact with the GABAA-receptor complex in an identical manner to the sedative barbiturate pentobarbitone. PMID- 2553193 TI - Evidence for tonic activation of prejunctional beta-adrenoceptors in guinea-pig pulmonary arteries by adrenaline derived from the adrenal medulla. AB - 1. The effects of (+/-)-carteolol 10(-8) M to 10(-6) M, a non-selective beta antagonist, applied cumulatively, on stimulation-evoked 3H-release at 1 Hz were studied in pulmonary arteries isolated from guinea-pigs. The guinea-pigs were subjected to either bilateral adrenalectomy, adrenalectomy followed by injections of deoxycorticosterone acetate (DOCA) and hydrocortisone, bilateral adrenodemedullation or a sham operation, and then loaded in vitro with [3H] noradrenaline. 2. Carteolol inhibited 3H-output in arteries from sham-operated animals in a concentration-dependent manner. This inhibitory effect was not found in pulmonary arteries from animals subjected to adrenalectomy or adrenodemedullation. However, DOCA and hydrocortisone pretreatment, did not prevent the disappearance of the carteolol-induced inhibition of 3H-release. 3. Adrenalectomy and adrenodemedullation depleted or markedly reduced the endogenous contents of adrenaline in pulmonary arteries without altering the levels of dopamine and noradrenaline. 4. It is concluded that adrenaline, mainly derived from the adrenal medulla, acts as an endogenous agonist for tonically functioning prejunctional beta-adrenoceptors in guinea-pig pulmonary arteries, probably by being taken up and co-released with noradrenaline. PMID- 2553194 TI - Ventricular fibrillation is reduced in hypothyroid rats with enhanced myocardial alpha-adrenoceptor responsiveness. AB - 1. The severity of ventricular arrhythmias induced by coronary artery occlusion and reperfusion has been examined in control rats and animals made hypothyroid by pretreatment with 6-propylthiouracil (PTU). The maximal driving frequency and sensitivity of isolated left atria and papillary muscles to isoprenaline and to phenylephrine in the presence of propranolol, were also examined in tissues from control and hypothyroid animals. 2. Pretreatment with PTU resulted in a potentiation of responses to the alpha-adrenoceptor agonist phenylephrine in both left atria and papillary muscles, while responses to isoprenaline were depressed in left atria but unaltered in papillary muscles from hypothyroid animals. 3. In rats subject to coronary artery occlusion, PTU pretreatment reduced the incidence of ventricular fibrillation during acute myocardial ischaemia and abolished reperfusion-induced ventricular fibrillation. Mortality during myocardial ischaemia and reperfusion was also abolished. Diastolic blood pressure was similar in hypothyroid and control animals, but there was a small reduction in systolic blood pressure and a marked decrease in heart rate in PTU pretreated animals. 4. These results demonstrate that PTU-induced hypothyroidism represents a condition where cardiac alpha-adrenoceptor-mediated responses are enhanced but the severity of ischaemia- and reperfusion-induced arrhythmias is reduced. PMID- 2553195 TI - Inhibitory effect of pyrophosphate, citrate, magnesium and chondroitin sulphate in calcium oxalate urolithiasis. AB - The inhibitory capacity of pyrophosphate, citrate, magnesium and chondroitin sulphate was investigated, using the urine of 21 calcium oxalate stone-forming patients without metabolic alterations. The inhibitory effect of these substances was assessed by a combination of nephelometry (light scattering) and optical microscopy. The results showed that citrate and magnesium had an inhibitory effect in a significant number of cases. Pyrophosphate and chondroitin sulphate had a less marked effect. The main urinary lithogenic biochemical parameters of the patients were also studied to see if there was a relationship between them and the inhibitory capacity of the compounds. PMID- 2553196 TI - Factors influencing local recurrence after excision and radiotherapy for primary breast cancer. AB - Between November 1979 and December 1986, 263 patients were treated for primary breast cancer by local excision and radiotherapy at the City Hospital, Nottingham. Local recurrence within the treated breast occurred in 56 patients (21 per cent), in 18 (6.8 per cent) of whom it was gross and uncontrollable. An analysis of clinicopathological features shows patient age, nodal status, tumour size, presence of definitive vascular invasion, adjacent ductal carcinoma in situ and grade to be predictive of local recurrence. A Cox's multivariate analysis of these factors shows the first four to be independently significant. The factors can be combined as a prognostic index which allows identification of patients at high risk of local recurrence. On the basis of these findings we have altered our selection policy for patients suitable for breast conservation. PMID- 2553197 TI - Changing patterns in gastric adenocarcinoma. AB - A retrospective study was performed on gastric cancer patients admitted to the Cancer Institute Hospital, Tokyo, during the periods 1961-65 (n = 1181) and 1980 84 (n = 1473). The aim of the study was to see whether changes had occurred in gastric cancer patients over the years with respect to histology and tumour localization and, if so, how they compared with reports from the West. The proportion of proximally localized primary tumour increased from 17 to 27 per cent (P = 0.008), and signet-ring cell carcinoma increased from 2 to 22 per cent (P less than 0.001). These changes were not relative with regard to the declining incidence of intestinal tumours, but absolute. Patients with proximally localized tumours were generally in a more advanced stage of disease than those with distally localized tumours, and thus had poorer survival rates. However, after stratifying into stages, the difference in survival disappeared. The survival rate of patients with signet-ring cell carcinoma was not significantly different from that of patients with other gastric tumours (P greater than 0.05). The discrepancy between the survival rates of Japanese and western gastric cancer patients is attributed by some authors to tumour-related factors. However, we believe that the similar trends found with regard to tumour localization and histology point towards comparable tumour behaviour in the two different geographical areas. PMID- 2553198 TI - Differential modulation of Ca2+-activated K+ channels in ovine pituitary gonadotrophs by GnRH, Ca2+ and cyclic AMP. AB - Patch-clamp techniques were employed to examine the effects of cAMP in relation to gonadotrophin-releasing hormone (GnRH) action on Ca2+-activated K+ channels in pituitary gonadotrophs derived from ovine pars tuberalis. GnRH applied extracellularly increased channel openings in cell-attached patches similar to calcium ionophore (A23187), while raising intracellular cAMP concentration with dibutyryl cAMP or forskolin decreased the number of functional channels (Nf) and the open state probability (Po). Both cAMP and the catalytic subunit of cAMP dependent protein kinase produced similar results when applied to the cytoplasmic membrane face of inside-out patches, and the effect of cAMP was abolished by the protein kinase inhibitor. Our results suggest that decreased permeability through these channels modulated by cAMP through a phosphorylation-dependent route can modulate luteinizing hormone release. PMID- 2553199 TI - Paraventricular nucleus injections of noradrenaline: cardiovascular effects in conscious Long-Evans and Brattleboro rats. AB - The cardiovascular effects of noradrenaline injected into the hypothalamic paraventricular nucleus (PVN) were investigated in conscious Long-Evans (control) rats and homozygous vasopressin (AVP)-deficient Brattleboro rats. Unilateral microinjection of noradrenaline (3-30 nmol) into the PVN produced dose-dependent increases in systolic and diastolic blood pressure of Long-Evans rats, and a concomitant decrease in heart rate. Only the highest dose of noradrenaline tested (30 nmol) caused a significant pressor response in Brattleboro rats (9 +/- 4/9 +/ 4 mm Hg, systolic/diastolic, n = 7) which was significantly smaller than the response produced by the same dose of noradrenaline in Long-Evans rats (32 +/- 8/27 +/- 6 mm Hg, n = 7). Intravenous pretreatment of Long-Evans rats with the V1 receptor antagonist, d(CH2)5Tyr[Et]DAVP, almost completely abolished the pressor effect of noradrenaline (10 nmol) without significantly attenuating the bradycardia. The alpha 2-adrenoceptor antagonist, idazoxan (4 nmol), injected into the PVN abolished the pressor response produced by noradrenaline (10 nmol) in Long-Evans rats but had no significant effect on the bradycardia. Pretreatment with the alpha 1-adrenoceptor antagonist, prazosin (0.7 nmol), significantly attenuated both the pressor and bradycardic effects of noradrenaline in Long Evans rats. These results suggest that the pressor response produced by microinjection of noradrenaline into the hypothalamic PVN of conscious Long-Evans rats is mediated largely through stimulation of alpha 2-adrenoceptors and is dependent, in part, on release of AVP into the circulation. A component of the bradycardia seen with this intervention may be mediated through stimulation of alpha 1-adrenoceptors in the PVN. PMID- 2553200 TI - Localization of calcitonin gene-related peptide and its receptors in a striated muscle. AB - The distribution of calcitonin gene-related peptide (CGRP) and its binding sites in the bulbocavernosus, a striated muscle, are reported. We used immunohistochemistry and [125I]CGRP autoradiography. The pattern of [125I]CGRP binding was restricted to a discrete band that coincides with the distribution of end-plates in this muscle as determined by CGRP immunohistochemistry and acetylcholinesterase staining. CGRP has been shown to increase the level of acetylcholine receptor (AChR) alpha-subunit mRNA. The role of CGRP as the endogenous factor by which motoneurons regulate the expression of junctional AChR is discussed. PMID- 2553201 TI - Modulation of the voltage-dependent sodium channel by agents affecting G proteins: a study in Xenopus oocytes injected with brain RNA. AB - The effects of agents known to affect G-proteins on voltage-dependent, tetrodotoxin-sensitive Na+ channels were studied in Xenopus oocytes injected with rat brain RNA, using two-electrode voltage-clamp technique. The non-hydrolysable analogue of GTP, GTP-gamma-S, known to activate G-proteins, inhibited the Na+ current (INa). The decrease in the amplitude of INa was not accompanied by changes in activation or inactivation characteristics of the channel. The non hydrolysable analogue of GDP, GDP-beta-S, had no effect on INa. The responses to gamma-aminobutyric acid and kainate in the same oocytes were also attenuated by GTP-gamma-S. Pertussis toxin, which inactivates some G-proteins by catalyzing their ADP-ribosylation, enhanced INa, but did not prevent the inhibition of INa by GTP-gamma-S. We conclude that the Na+ channel, and possibly the GABA and kainate receptors and/or channels, are coupled to a G-protein. The activation of the G-protein modulates the channels either directly, or via activation of biochemical cascade possibly involving production of second messengers and channel phosphorylation. PMID- 2553202 TI - Biochemical evidence for enhanced sensitivity to N-methyl-D-aspartate in the hippocampal formation of kindled rats. AB - The inhibitory effect of N-methyl-D-aspartate (NMDA) upon carbachol-stimulated phosphoinositide (PI) hydrolysis was studied in transverse hippocampal slices prepared from control and amygdaloid kindled rats. Kindling significantly increased the inhibitory effect of NMDA (10 microM) in slices prepared from animals 24 h after the last class 5 kindled seizure, resulting in a steepening of the dose-response curve for NMDA. The enhanced sensitivity to NMDA was long lasting in that it was also present in slices prepared from animals sacrificed 28 35 days after the last class 5 seizure. The increased sensitivity to NMDA was selective in that inhibition of carbachol-stimulated PI hydrolysis by kainic acid or phorbol-12,13-diacetate was not different in control and kindled animals. Neither NMDA, kainic acid, phorbol ester nor carbachol alone had any significantly different effects in slices from kindled versus control animals. These data demonstrate a selective and enhanced sensitivity of the kindled hippocampus to NMDA. This enhanced sensitivity to the principal class of excitatory neurotransmitter may be one mechanism underlying the development and maintenance of kindled epilepsy. PMID- 2553203 TI - Effects of NMDA antagonists, MK-801 and CPP, upon local cerebral glucose use. AB - The effects upon cerebral glucose utilisation of (+)-5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine (MK-801, a non-competitive N-methyl-D aspartate (NMDA, receptor antagonist) and 3-(2-carboxypiperazin-4-yl)-propyl-1 phosphonic acid (CPP, a competitive NMDA receptor antagonist) were examined in conscious, lightly restrained rats. Cerebral glucose utilisation was assessed quantitatively in 74 brain regions with [14C]2-deoxyglucose autoradiography. The intravenous (i.v.) administration of MK-801 (0.05-5 mg/kg) induced heterogeneous patterns of altered cerebral glucose utilisation with statistically significant increases being observed in 21 brain areas and statistically significant decrease in 8 brain regions. Pronounced dose-related increases in glucose use were observed after MK-801 in the subicular complex, hippocampus molecular layer, dentate gyrus, limbic system (posterior cingulate cortex; mamillary body; anteroventral thalamic nucleus), olfactory areas and substantia nigra (pars reticulata). Glucose use in the neocortex and inferior colliculus was particularly sensitive to reduction by MK-801 administration. The pattern of altered glucose use after administration of CPP (3-30 mg/kg, i.v.) differed markedly from that observed after MK-801 treatment. Statistically significant increases in glucose use after CPP were noted in 11 brain areas and statistically significant decreases in 5 of the regions examined. Regions in which increases were noted after CPP included hippocampus molecular layer, olfactory areas, cochlear nucleus, vestibular nucleus, cerebellar nucleus, superior olives and substantia nigra (pars reticulata). These data indicate that widespread, anatomically organised alterations in cerebral function are associated with the administration of NMDA receptor antagonists despite the minor role normally ascribed to these receptors in conventional fast synaptic transmission. The distinct patterns of response to competitive and non-competitive antagonists may be a reflection of the differential responses of the two modes of receptor blockade to increased glutaminergic transmission. PMID- 2553204 TI - [3H]imipramine binding in discrete brain areas is affected by castration in male rats. AB - In adult male rats, castration induces a progressive decrease in the number of [3H]imipramine binding sites in the cerebral cortex and hypothalamus, and a progressive increase in the hippocampus. Testosterone completely prevents this effect of castration, but has no effect on the characteristics of brain imipramine binding sites in intact, non-castrated animals. These data suggest that threshold levels of testosterone are necessary for the maintenance of a normal number of imipramine binding sites in the rat brain, but that these binding sites are not modified by excess testosterone. PMID- 2553205 TI - Inhibition of histamine release from rat hypothalamic slices by omega-conotoxin GVIA, but not by nilvadipine, a dihydropyridine derivative. AB - Histamine release in response to 40 mM high K+-stimulation from the rat hypothalamic slice preparations perifused in vitro was significantly inhibited by 1.0 nM-1.0 microM omega-conotoxin GVIA, a peptide modulator of N- and L-type voltage-sensitive calcium channels, but not by similar concentrations of nilvadipine, a dihydropyridine derivative of L-type calcium channel antagonist. These results indicate that the voltage-sensitive calcium channel controlling histamine release from hypothalamic slices is omega-conotoxin-sensitive but dihydropyridine-insensitive. PMID- 2553206 TI - Beta-adrenergic receptors in the vasopressin-containing neurons in the paraventricular and supraoptic nucleus of the rat. AB - Immunocytochemical staining of alternate consecutive sections revealed that neurons with vasopressin-like immunoreactivity in the paraventricular and supraoptic nuclei had beta-adrenergic receptor-like immunoreactivity in the rats. PMID- 2553207 TI - Distribution of [3H]MK801 binding in the normal aged human hippocampus. AB - The distribution of NMDA receptors in the normal human hippocampus has been investigated using the non-competitive channel blocking agent, MK801. The pattern of specific [3H]MK801 binding was broadly similar to that previously described for agonist binding although differences included equal densities in h1 and h2 regions where binding was mainly confined to the pyramidal layer and stratum radiatum. PMID- 2553208 TI - Changes of salt intake and of (Na+K)-ATPase activity in brain after high dose treatment with deoxycorticosterone. AB - Mineralocorticoids (MC) have a dual effect on salt intake: in adrenalectomized rats, they reduce previously elevated salt intake; and in intact rats a high MC dose increases salt intake. We have studied the activity of (Na+K)-ATPase and [3H]ouabain binding in rats treated with deoxycorticosterone (DOC) in doses that elicited a salt appetite. Brains were removed from control and treated animals, and 20 different areas were punched out from brain slices cut every 300 microns. DOC treatment significantly reduced (Na+K)-ATPase activity in the lateral hypothalamic area, anterior amygdaloid and lateral amygdaloid nuclei, while increasing it in the periventricular gray matter; changes in other regions were not significant. Binding of [3H]ouabain was not modified by DOC treatment. In parallel experiments, we determined MC receptors in adrenalectomized rats. Binding of [3H]aldosterone was preferentially found in hippocampus, followed by lateral septum, anterior, posterior and lateral amygdaloid areas, with lower levels in other regions. However, there was no correlation between [3H]aldosterone binding and (Na+K)-ATPase activity in brain punches from either control or DOC-treated rats. Further experiments are needed to ascertain if (Na+K)-ATPase changes in discrete areas of the brain containing moderate levels of mineralocorticoid receptors, are related to the behavioral effects of DOC. PMID- 2553209 TI - Activation of N-methyl-D-aspartate (NMDA) receptors augments repolarizing responses in lamprey spinal neurons. AB - Current- and voltage-clamp techniques were used to analyze the mechanisms underlying the repolarization during N-methyl-D-aspartate (NMDA)-induced, tetrodotoxin-resistant pacemaker-like oscillations in lamprey spinal neurons. Long-lasting depolarizing current pulses (15-40 mV, 50-400 ms, tetrodotoxin and tetraethylammonium present) were followed by hyperpolarizing afterpotentials even when NMDA receptors were blocked, but they were markedly enhanced by application of N-methyl-D,L-aspartate (NM(DL)A). The afterpotentials were depressed by replacing Ca2+ with Ba2+. During voltage-clamp NM(DL)A enhanced a Ba2+-sensitive outward tail current following voltage steps of 15-40 mV. The outward current remained after injection of Cl-, as did the NMDA-induced membrane potential oscillations observed under current-clamp. These results suggest that the repolarization during NMDA-induced oscillations is due to Ca2+ entry both via NMDA-gated channels and conventional voltage-gated Ca2+ channels, leading to an activation of Ca2+-dependent K+ channels. The afterhyperpolarization following single action potentials, which is also due to Ca2+-dependent K+ channels, was not significantly altered by NMDA receptor activation, suggesting a different location of the Ca2+ entry during the two conditions in relation to the location of the activated Ca2+-dependent K+ channels. PMID- 2553210 TI - Preferential loss of GABAergic neurons in hypoxia-exposed neocortex slab cultures is attenuated by the NMDA receptor blocker D-2-amino-7-phosphonoheptanoate. AB - The preferential loss of GABAergic neurons in neocortex slab cultures after exposure to hypoxia was reinvestigated and confirmed using an improved morphometric analysis based on stereological principles. This preferential cell loss could be attenuated by treating the hypoxic slab cultures with the N-methyl D-aspartate (NMDA) receptor blocker D-2-amino-7-phosphonoheptanoate (D-APH). PMID- 2553211 TI - Degeneration of neurons in the thalamic reticular nucleus following transient ischemia due to raised intracranial pressure: excitotoxic degeneration mediated via non-NMDA receptors? AB - Transient global ischemia was produced in rats by cisternal fluid infusion, producing a negative cerebral perfusion pressure by elevating the intracranial pressure (ICP) 25-50 mm Hg above mean arterial pressure (MAP). Animals were allowed to survive for 2-7 days following a transient ischemic episode of 5-30 min. The brains were examined for signs of ischemic degeneration in Nissl-stained sections and adjacent sections reacted with antisera against glial fibrillary acidic protein (GFAP) or aspartate aminotransferase (AAT). Neurons in the thalamic reticular nucleus (RT), a pure population of gamma-aminobutyric acid (GABA)ergic neurons which project their axons to thalamic relay nuclei, were found to have the lowest threshold for degeneration in this model, consistently undergoing degeneration under conditions which completely spared the hippocampal CA1 from degeneration. Whereas it took up to 30 min of complete ischemia to produce degeneration of CA1 neurons when ICP was raised using room temperature infusion fluids, 15 min of ischemia under these conditions was sufficient to produce extensive degeneration of neurons in the entire ventral 3/4 of the RT. Prolonged (greater than 25 min) episodes of partial ischemia (ICP less than or equal to MAP) were also sufficient to produce massive degeneration of RT neurons. The lesion in the RT was most clearly evident in sections reacted with antisera to GFAP, labeling intensely reactive protoplasmic astrocytes within the regions of the RT where neuronal degeneration had occurred. Neuronal loss and accompanying proliferation of microglial cells were evident in Nissl-stained sections but the extent of the neuronal loss was most clearly obvious in sections reacted with an antisera to AAT, an enzyme present in detectable quantities in GABAergic neurons. Pretreatment with the non-competitive NMDA antagonist MK-801 at doses sufficient to completely prevent massive degeneration of the hippocampal CA1 failed to prevent the degeneration of RT neurons, suggesting that if RT degeneration involves an excitotoxic process it acts through non-NMDA receptors. PMID- 2553212 TI - Dramatic increase in nigral t-[35S]butylbicyclophosphorothionate binding sites elicited by the degeneration of the striato-nigral GABAergic pathway: reversal by diazepam. AB - In rats, the degeneration of the striato-nigral GABAergic pathway caused by the intrastriatal injection of kainic acid induced a marked decrease (65%) of GABA content and glutamic acid decarboxylase (GAD) activity and a dramatic increase (225%) in the binding of t-[35S]butylbicyclophosphorothionate [( 35S]TBPS) to a membrane preparation from the substantia nigra homolateral to the injected striatum. The increase in [35S]TBPS binding in the denervated substantia nigra was exclusively due to an increased density of binding sites (Bmax) with no change in the dissociation constant (kd). The enhancement in [35S]TBPS binding was almost completely reversed by the intraperitoneal administration of diazepam (3 mg/kg) to kainic acid-lesioned rats. Moreover, diazepam produced a significant decrease (30%) in the density of [35S]TBPS binding sites also in the sham operated side. In contrast the 'in vitro' addition of the GABAA receptor antagonist bicuculline (1 microM) to the membrane preparation from the denervated substantia nigra further increased [35S]TBPS binding. These findings suggest the view that the increase of nigral [35S]TBPS binding is directly related to the inhibition in the function of nigral GABAergic synapses following the loss of the striato-nigral GABAergic pathway. Our results indicate that [35S]TBPS binding to brain structure is a potential tool to reveal alteration in the function of GABAA receptor complex elicited by physiological, pharmacological and pathological conditions. PMID- 2553213 TI - Relationship of calcium and adenylate cyclase messenger systems in rat brain synaptosomes. AB - The effects of cyclic AMP on the rise in cytosolic free calcium concentration, [Ca2+]i, after stimulation with 15 mM K+ in rat brain synaptosomes were investigated. The fluorescent chelating agent Quin-2 was employed to monitor alterations of K+-evoked [Ca2+]i. Under normoxic conditions, clonidine (1, 10 microM), an alpha 2-adrenoceptor agonist, decreased the 15 mM K+-evoked [Ca2+]i. Although yohimbine (1, 10 microM), an alpha 2-adrenoceptor antagonist, had little or no effect on K+-evoked [Ca2+]i, the inhibitory effects of clonidine were blocked by yohimbine. 8-Bromo cyclic AMP, a cyclic AMP analogue, (50-500 microM), increased K+-evoked [Ca2+]i in a dose-dependent manner. The addition of cyclic AMP analogues subsequent to clonidine treatment reversed the clonidine-induced suppression of K+-evoked [Ca2+]i. On the other hand, under hypoxic conditions, K+ evoked [Ca2+]i was reduced by about 50-60%. 8-Bromo cyclic AMP and the adenylate cyclase activators, yohimbine (1-10 microM) and isoproterenol, a beta adrenoceptor agonist, (0.1-10 microM), transiently reversed the reduction of the K+-evoked [Ca2+]i caused by hypoxia. These results indicate that the activation of alpha 2-adrenoceptor produces a rapid, sustained decrease in [Ca2+]i which may be due to a decrease in the levels of intracellular cyclic AMP. In addition, the increase in cellular levels of cyclic AMP reversed the reduction of the Ca2+ response to high K+ stimulation caused by hypoxia. If this is so, there is the possibility that increased cyclic AMP might improve the hypoxic damage. PMID- 2553214 TI - Ouabain (or salt solution lacking potassium) mimics the effects of dark pulses on the circadian pacemaker in cultured chick pineal cells. AB - Chick pineal cells in static culture display a persistent photosensitive circadian rhythm of melatonin production and release. Pulses of white light or darkness, in otherwise constant red light, induce phase shifts in subsequent cycles whose magnitude and direction depend on the phase at which the pulse is given. Such 'phase-dependent phase shifts' are mediated by effects on the underlying pacemaker. Here, we describe the effects of ouabain, a specific inhibitor of Na,K-ATPase, and of salt solutions lacking potassium (SS-K), which also inhibit the pump, on the melatonin rhythm. Pulses of ouabain, or of SS-K, induced phase advances and phase delays that were phase and concentration dependent. The relationship between time of treatment and effect on the subsequent phase of the rhythm (the phase-response curve) for these treatments was essentially the same as that for pulses of darkness. PMID- 2553215 TI - Atrial natriuretic polypeptide attenuates central angiotensin II-induced catecholamine and ACTH secretion. AB - The interaction between angiotensin II (AII) and synthetic atrial natriuretic polypeptide (ANP) on the sympathetic nervous system and ACTH secretion was examined in unanesthetized, freely moving rats. Centrally administered AII (100 ng/2 microliters) caused hypertension with elevation of the plasma epinephrine level. Central administration of ANP (3 micrograms/3 microliters, 10 micrograms/3 microliters) attenuated central AII-induced pressor response and plasma epinephrine elevation. Furthermore, central AII stimulated ACTH secretion, and ANP reduced the ACTH secretion induced by AII. These results show that ANP attenuates the central AII-induced pressor response at least partially by suppressing sympathetic nervous activity, and ANP regulates ACTH secretion at the hypothalamic level. PMID- 2553216 TI - Localization studies of gamma-hydroxybutyrate receptors in rat striatum and hippocampus. AB - Quantitative autoradiography using [3H] gamma-hydroxybutyrate was used in combination with anatomic and neurotoxic lesions to localize the gamma hydroxybutyrate (GHB) receptors in the striatum and hippocampus of rat brain. 6 Hydroxydopamine (6-OHDA) lesions of the nigro-striatal pathway failed to reduce [3H] gamma-hydroxybutyrate binding in the striatum. In contrast, kainic acid (KA) lesions of the caudate-putamen (CPu) resulted in about 45% loss of binding. For hippocampus, lesions of septo-hippocampal pathway did not modify receptor density but intrahippocampal kainic acid injection largely attenuated (50%) [3H] GHB binding. These results demonstrate that gamma-hydroxybutyrate receptors in the CPu and dorsal hippocampus are principally located on intrinsic neurons which may participate in the functional expression of the role gamma-hydroxybutyrate has in these structures. PMID- 2553217 TI - MK-801 protects against seizures induced by the cholinesterase inhibitor soman. AB - MK-801, a novel anticonvulsant which is a potent N-methyl-d-aspartate antagonist, attenuated or blocked seizures and convulsions induced by the irreversible organophosphorus anticholinesterase inhibitor soman. Guinea pigs chronically instrumented for electrocorticogram recording were given a low dose (1 mg/kg) or a high dose (5 mg/kg) of MK-801 or saline vehicle 30 min prior to receiving 2 x LD50 of soman. All animals were treated with atropine methylnitrate and pralidoxime chloride 30 sec after soman injection. All saline control subjects exhibited severe seizures and convulsions shortly after receiving soman. Low dose MK-801 greatly attenuated and high dose MK-801 completely blocked seizure activity. Animals treated with MK-801 recovered faster and had a much greater probability of survival for 48 hr after soman exposure than did controls. This is the first demonstration of the involvement of the excitatory amino acid neurotransmitter system in seizures and convulsions induced by a cholinesterase inhibitor. PMID- 2553218 TI - Interaction of ethanol and stress with the GABA/BZ receptor in LS and SS mice. AB - The interaction of stress and ethanol with the GABA/BZ receptor system was evaluated in LS and SS mice. The effects of two separate in vivo treatments, a 2.5 g/kg injection of ethanol or a behavioral stressor, on GABA-enhanced [3H]-FNZ binding were nearly identical in both lines of mice. A 2.5 g/kg ethanol- or stress-pretreatment resulted in increased enhancement in SS cortex, but not LS. In cerebellum, treatment effects were demonstrated in both SS and LS mice. Intraperitoneal injections of increasing doses of ethanol produced biphasic stimulation of GABA-enhanced [3H]-FNZ binding in LS brain regions, but not SS. Adrenalectomies performed one week prior to ethanol administration produced a loss of ethanol enhancement in cerebellum of both lines. However, in cortex, removal of the adrenals had no effect. The in vitro addition of 30 mM ethanol to brain preparations incubated at 37 degrees C from stressed and unstressed animals resulted in greater enhancement of binding in cortex, but not cerebellum of stressed mice. Differences in the degree of enhancement between the lines of mice were lost if the animals were stressed prior to sacrifice or if membrane preparations were incubated at 4 degrees C. The results of this study suggest that the interaction between ethanol and stress is mediated by the GABAergic system, but responses vary dependent on brain region, dose of ethanol, and degree of ethanol sensitivity. PMID- 2553219 TI - [Cat retroviruses and human retroviruses: elements of comparison]. AB - Following emotional head-lines of certain articles in the press, making believe that the cat could be susceptible to the AIDS virus, the authors present elements of comparison between principal feline retroviruses (the feline leucosis virus and the feline immunodeficiency virus) and the two human immunodeficiency viruses (HIV). The feline leucosis virus in differentiated from the human and the feline immunodeficiency viruses by its virological, pathological and epidemiological characteristics. Being close to the AIDS virus in the taxonomy of retroviruses, the feline immunodeficiency virus (FIV) presents a number of similarities with the HIV. Therefore, the FIV could give rise to interests in its use as a model in the study of AIDS. Whatever the factors of resemblances may be, there are no elements of present knowledge in favor of an inter-species contamination (cat man); on the contrary, these viruses demonstrate a marked species specificity. PMID- 2553220 TI - A continuous line of chicken embryo cells derived from a chondrocyte culture infected with RSV. AB - A continuous line of chicken embryo cells was derived from a culture of chondrocytes infected with Rous sarcoma virus (RSV). These cells, designated as NA101, have reduced serum requirements and are able to grow in a semisolid medium. NA101 cells show the same phenotype as freshly RSV infected chondrocyte cultures, i.e. synthesis of fibronectin and type I collagen, elongated bipolar shape and absence of tumorigenicity following grafting onto the chorioallantoic membrane of embryonated duck eggs; thus they appear to be distinct from transformed fibroblasts. Neither NA101 cells nor freshly infected chondrocyte cultures synthesize type II or type X collagen, which are the differentiation markers of normal chicken chondrocytes in culture. PMID- 2553221 TI - [Human papillomavirus infection of the cervix uteri in married women]. PMID- 2553222 TI - [Endonasal removal of pituitary growth hormone-adenoma: report of 56 cases]. AB - Fifty-six cases of pituitary GH-adenoma removed by endonasal method are reported. In 50 cases, plasma hGH decreased postoperatively. According to biological evaluation, 18 patients were cured, 21 were improved and 17 presented no change. Postoperative high hGH (greater than or equal to 5 ng/ml) indicated that there were some residual functioning tumor cells in the Sella. Such cases must be followed up regularly in order to assess when and what treatment should be given. Postoperative radiation usually raises the curability of acromegaly. Endonasal method is simple, effective and less injurious to the neighboring structure of the nose. PMID- 2553223 TI - Biochemical characterization of a virus-induced osteosarcoma-like osseous lesion in vitro. AB - Chondroprogenitor cells present in the apical and lateral parts of the mandibular condyle from neonatal mice differentiate towards the osteoblastic lineage and form bone within 7 days in culture. Infection of condylar explants with the FBR osteosarcoma virus (FBR MSV) results in the transformation of cells in the progenitor zone, previously identified as the target for the virus, and the formation of a transplantable osteosarcoma-like lesion. Morphological and biochemical changes in this system were investigated in the course of tumor development. Virus infection was followed by a significant increase in cell density and 3H-thymidine incorporation within the progenitor zone at the early stage of culture. In later stages, cell density and 3H-thymidine incorporation were lower than in control tissue. The 3H-thymidine labeling index gave similar results in infected and control tissues until day 7. Then, a significantly higher labeling index was found in the progenitor zone of infected condyles. At this stage, the proliferative effect of the virus even affected the cartilagenous core of the tissue. Quantitative alkaline phosphatase activity increased between day 3 and day 7 and was particularly high in the zone of infected cells. In addition, infected tissues consistently revealed a higher uptake of 45Ca, and deposition of the radioisotope along irregularly formed bone trabecules in the transformed tissue. The results suggest that there is an enhancement of tissue maturation following infection with the FBR osteosarcoma virus. Although biochemical investigations of whole condyles showed few differences in the total values of alkaline phosphatase activity, 3H-thymidine incorporation, DNA content, and 45Ca uptake, the histochemical assays revealed clear differences in the distributional pattern of these parameters within infected and control condyles. PMID- 2553224 TI - Neurotransmitter regulation of cytosolic calcium in osteoblast-like bone cells. AB - The nervous system may play a role in regulation of bone metabolism. The effects of norepinephrine(NE), vasoactive intestinal peptide(VIP), and ATP on cytosolic Ca2+ were assessed in a rat osteoblast-like osteosarcoma cell line (UMR-106) responsive to PTH. All three transmitters transiently increased Ca2+, with ATP much greater than PTH greater than NE = VIP, and then caused sustained increases in Ca2+. The ATP-induced transient resulted from mobilization of intracellular Ca2+ store, while NE and VIP-induced transients also involved influx of Ca2+. Later sustained increases by all agonists were dependent upon extracellular Ca2+. Release of intracellular Ca2+ by ATP was associated with a marked increase in IP3 but without a significant change in cAMP. NE, VIP, and ATP, through regulation of Ca2+ metabolism, may be involved in various osteoporotic conditions. PMID- 2553225 TI - Evidence for in vivo upregulation of 1,25(OH)2 vitamin D3 receptor in human monocytes. AB - Mammalian cells increase net expression of 1,25(OH) 2D3 receptors after exposure to physiological concentrations of 1,25(OH) 2D3 in vitro. We examined specific binding of 1,25(OH) 2D3 by human monocytes before and after daily administration of 1.5-2 micrograms 1,25(OH) 2D3 p.o. for 3 days in 5 healthy normal D-replete probands. Median specific binding (Nmax) at baseline was 793 molecules/cell and 2052 or 2828 at 24h and 72h of 1,25(OH) 2D3 treatment respectively. The results suggest (a) upregulation of 1,25(OH) 2D3 receptors occurs in man and (b) monocyte preparations can be used to assess receptor regulation in vivo. PMID- 2553227 TI - [Changes in cAMP and cGMP in the liver and heart in early stages of burn injury in rats]. PMID- 2553228 TI - Combined modality treatment for non-small cell lung cancer. Lung Cancer Study Group. AB - A total of 265 patients with non-small cell lung cancer at stages 1 and 2 by preoperative clinical staging were divided into 4 stages after exploratory thoracotomy. Patients at each stage were further divided into subgroups to receive different adjuvant therapies. The cumulative 5-year survival rate showed to be 44.4% for stage 1, 35.5% for stage 2, 7.4% for stage 3 and 0% for stage 4. Histologically, the rate was 28.8% for squamous cell carcinoma, 23.4% for adenocarcinoma, and 29.8% for large cell carcinoma and carcinoma poorly differentiated. For patients at stage 1, it made no significant difference whether they underwent surgical resection only or combined with postoperative intrapleural BCG injection. For patients at stage 2, there was no significant difference between surgical resection only and resection combined with postoperative radiotherapy. For patients at stage 3, who underwent surgical resection combined with chemotherapy and/or radiotherapy, it seemed that combined chemotherapy and radiotherapy presented better results than radiotherapy or chemotherapy alone. PMID- 2553226 TI - Establishment from mouse growth cartilage of clonal cell lines with responsiveness to parathyroid hormone, alkaline phosphatase activity, and ability to produce an endothelial cell growth inhibitor. AB - Three clonal cell lines with differences in responsiveness to parathyroid hormone (PTH), alkaline phosphatase activity, and ability to produce an endothelial cell growth inhibitor(s) during more than 3 years, more than 58 passages, in culture were established from growth cartilage (GC) of mouse ribs. In sparse cultures the three clonal cell lines, MGC/T1.4, MGC/T1.17, and MGC/T1.18, all showed fibroblast-like morphology. However, as they became confluent, MGC/T1.4 cells became polygonal and then multilayered. MGC/T1.18 cells also became polygonal, but showed contact inhibition. MGC/T1.17 cells remained fibroblastic in confluent cultures and formed nodules when cultured for more than 7 days after they became confluent. These nodules calcified in the presence of beta-glycerophosphate. Glycosaminoglycan (GAG) synthesis in the parent uncloned line, MGC/T1 cells, at early passages was about 50-75% of that of primary cultures of mouse GC cells. The GAG syntheses in the three clonal lines were much lower than that of primary cultures of GC cells. Moreover, the sizes of proteoglycan monomers synthesized by these cells were not the same as that of cartilage-specific proteoglycan. The three clonal lines mainly synthesized type I collagen. PTH increased the intracellular cyclic AMP level in MGC/T1, MGC/T1.4, T1.17, and T1.18 cells: their maximal levels, observed after 2 minutes, were, respectively, about 160, 150, 70, and 200 times that of controls. The activity of alkaline phosphatase in MGC/T1.17 cells was higher than that in primary cultures of mouse GC cells, whereas those in MGC/T1 and T1.4 cells were comparable with that of GC cells, and that in MGC/T1.18 was lower. The three clonal lines, and especially MGC/T1.4, secreted a heat-stable, nondializable growth inhibitor(s) of endothelial cells into the culture medium. Because of their different properties, these cell lines should be useful for studies on endochondral ossification, the actions of PTH on skeletal cells, and anti-angiogenesis factors. PMID- 2553229 TI - Inclusion body myositis associated with progressive dysphagia: treatment with cricopharyngeal myotomy. AB - A 68-year-old man known to have inclusion body myositis underwent a cricopharyngeal myotomy in an attempt to improve his progressive dysphagia. Morphological studies from tissues obtained during this procedure showed the diagnostic features typical of this chronic inflammatory myopathy. To our knowledge this is the first pathological demonstration of inclusion body myositis involving the pharyngeal skeletal musculature. PMID- 2553230 TI - Nuclear inclusions in oculopharyngeal muscular dystrophy in Quebec. AB - Seven French-Canadian cases of clearcut oculopharyngeal muscular dystrophy (OPMD) had their muscle studied for the presence of intranuclear inclusions, and they were all positive. Inclusions of both "mature" and "immature" types were seen in our material. The presence of such intranuclear structures should be added to the criteria of the clinical picture and the family history for diagnosis of a case and inclusion of a family in further genetic studies. Reverse genetic studies of large families and biochemical studies of these intranuclear structures may help to understand the pathogenesis of this common disease in Quebec. PMID- 2553231 TI - Ultrastructural studies of the cells forming amyloid fibers in classical plaques. AB - Three-dimensional reconstruction and ultrastructural studies of classical plaques from the cortex of patients with Alzheimer's disease showed that microglial cells of the plaques are the amyloid-forming cells. The amyloid star of the single plaque represents the product of five or six microglial cells covering about 80% of the amyloid star surface. The amyloid fibers appear to be formed within altered cisterns of the endoplasmic reticulum. Distended cisterns form channels filled with amyloid fibers. Numerous vesicles derived from the Golgi apparatus appear to be attached to or fused with the amyloid-filled channels. Reconstruction of the amyloid star and the microglia cell pole that forms the amyloid star reveals three different zones of distribution of cytoplasmic organelles and amyloid deposits. The peripheral zone comprises channels filled with loosely packed amyloid fibers arranged in a parallel manner. The transient zone consists of a mixture of fusing amyloid channels and products of disintegration of cytoplasmic pockets, dense bodies and fragments of cellular membranes. The core of the amyloid star is composed of condensed, densely packed amyloid fibers that are free of cellular debris. Formation of the three zones supports the idea that the microglia/macrophages are not phagocytes but instead are the cells manufacturing the amyloid fibers. PMID- 2553232 TI - The relationship between Alzheimer's disease, Parkinson's disease and motor neuron disease. AB - We argue against the dominant status assigned to conventional microscopy in the categorization of disorders such as Alzheimer's disease, Parkinson's disease and ALS. As an example we criticize the emphasis that has been placed on correlating the presence of Lewy bodies with the diagnosis of Parkinson's disease. In essence, we submit that Parkinson's disease can exist without Lewy bodies, and Lewy bodies can exist without Parkinson's disease. Nevertheless, we consider that the newer techniques available to histology have led to an important concept that constitutes a shared feature for Alzheimer's disease. Parkinson's disease and ALS; they are all characterized by the deposition of cytoskeletal debris in tissue, so they may perhaps be collectively termed the "Cytoskeletal Disorders". PMID- 2553233 TI - Proportion of new AIDS case reports attributable to the 1987 CDC revised surveillance case definition: Canada-United States differences. PMID- 2553234 TI - Adjuvant intraarterial chemotherapy with nimustine in the management of World Health Organization Grade IV gliomas of the brain. Experience at the Department of Neurosurgery of Dusseldorf University. AB - A protocol with postoperative adjuvant intraarterial (ia) chemotherapy using nimustine (ACNU) was followed at the University of Dusseldorf (Federal Republic of Germany) after histologic confirmation of malignant glioma of the brain. The first iaACNU procedure was performed within 10 days of surgery, and immediately before postoperative irradiation. After radiotherapy further iaACNU courses were given. The study population consisted of 35 primarily treated malignant gliomas World Health Organization (WHO) Grade IV, i.e., glioblastomas (NIV). Of these tumors 11 showed early progressive disease (PD) (NIV primPD), whereas 24 had stable or responsive disease (NIV SD&RD). Seven patients with recurrent malignant gliomas WHO Grade IV (NIV recurrent) were also treated plus six patients with malignant gliomas WHO Grade III (NIII). Experience with 50 treated patients resulted from this study. The median survival time (MST) for the group NIV was 14.2 months; 41.0% of patients were long-term survivors. The MST for the group NIV primPD was 7.8 months; 9.1% of patients were long-term survivors. For the group NIV SD&RD a MST of 18.3 months and 66.7% of long-term survivors were determined. Patients with recurrent tumors on this treatment protocol had an additional MST of 6.1 months. The total MST after first surgery was 16.2 months. The incidence of complications was low. Systemic complications were infrequent or rare. Retinal disturbances were seen in two patients; leukoencephalopathy was not seen in our patients. In 10% of patients cerebral complications were found, which were irreversible in 2% of patients. Complications could not be assigned unequivocally to the nitrosourea. These results compare favorably with what is reported in the literature, demonstrate the potential value of iaACNU in the treatment of malignant gliomas WHO Grade IV, and should encourage the initiation of a randomized clinical trial of adjuvant iaACNU in the therapy of these tumors. The authors propose iaACNU as a valuable approach in the management of primarily diagnosed malignant gliomas WHO Grade IV, but cannot recommend the delay of iaACNU until the recurrence of a brain tumor. PMID- 2553235 TI - Weekly doxorubicin versus doxorubicin every 3 weeks in cyclophosphamide, doxorubicin, and cisplatin chemotherapy for non-small cell lung cancer. AB - A prospective randomized study was done to determine the effect of different doxorubicin (Adriamycin [ADR], Adria Laboratories, Columbus, OH) administration (schedules every week versus every 3 weeks) on the productivity of a cyclophosphamide, ADR, cisplatin (CAP) chemotherapy regimen for patients with non small cell lung cancer (NSCLC). Electrocardiograms, multigated cardiac scans, echocardiograms, and endomyocardial biopsies were done serially for cardiac monitoring. Of 102 patients, 47 ahd inoperable limited disease (LD), 47 had extensive disease (ED), and eight had no evidence of disease. In the last group chemotherapy was given adjuvantly. Fifty-one patients were entered into each treatment arm. The groups were formed according to extent of disease and were comparable in terms of patient characteristics. In these groups, the overall response rates using both schedules in LD patients were similar: in patients without chest irradiation previously, there was a response of 35% with ADR weekly, and 31% with ADR triweekly; in LD patients with chest irradiation previously, the response was 20% with ADR weekly, and 25% with ADR triweekly; and in ED patients, 16% with ADR weekly, and 11% with ADR triweekly. There was no significant difference in survival between the two treatment groups. However, results for all responders suggested a longer duration of response with weekly than with triweekly ADR (complete plus partial response: 35.8 versus 11.4 weeks, P = 0.06; minor response: 34 versus 11.5 weeks, P = 0.003, respectively). Results also suggested that weekly ADR was less cardiotoxic than triweekly ADR: 29% of patients in the former group had no changes or only minor changes in endomyocardial biopsy results, whereas all patients in the latter group had at least grade 0.5 changes at a similar dosage. The median doses of weekly ADR were higher at the same endomyocardial biopsy-defined toxicity levels. No correlation was found between toxic effects defined by endomyocardial biopsy results and those defined by noninvasive monitoring techniques, although the number of patients assessed was small. Weekly ADR produced less granulocytopenia and a lower incidence of fever (6% versus 16%, P less than 0.001) than did triweekly ADR. Alopecia, nausea, vomiting, and diarrhea were significantly less for weekly ADR than triweekly Adr (P less than 0.0005, less than 0.0005, and less than 0.005, respectively). These data suggest that weekly ADR can achieve the same therapeutic results as the standard triweekly regimen with less cardiotoxicity, myelotoxicity, alopecia, diarrhea, nausea, and vomiting in patients with NSCLC. PMID- 2553236 TI - Intraarterial infusion chemotherapy for metastatic liver tumors using multiple anti-cancer agents suspended in a lipid contrast medium. AB - An ethiodized oil, Lipiodol (Lipiodol Ultra-Fluid, Laboratoire Guerbet, Paris, France), when injected into the hepatic artery, is selectively retained by liver tumors. Thus, anti-cancer agents suspended in Lipiodol can be delivered specifically to liver tumors. Before clinical trials of this new drug delivery system were begun, the movements of drugs suspended in Lipiodol were examined in vitro. It was found that 5-fluorouracil, doxorubicin, and mitomycin C were continuously released from oil phase to water phase, and when these three drugs were collectively suspended in Lipiodol, each was released independently. During clinical investigations, 42 patients with unresectable metastatic liver tumors were treated with intraarterial infusions of anti-cancer drugs in Lipiodol. As a result, the administration of a combination of anti-cancer agents suspended in Lipiodol was shown to be effective in controlling metastatic liver tumors. It was concluded that such a method could be applied to a variety of metastatic liver tumors with different drug sensitivities. PMID- 2553237 TI - Complete remission of refractory small cell carcinoma of the pancreas with cisplatin and etoposide. AB - A case of unusually large widespread metastatic small cell carcinoma of the pancreas is presented. The patient remains in complete remission 50 months after the diagnosis and initiation of combination chemotherapy using a schedule consisting of etoposide (150 mg/m2) intravenously (IV) on days 1 to 3 and cisplatin (100 mg) IV on day 2. Initial chemotherapy with streptozotocin, 5 fluorouracil, and methotrexate, doxorubicin, cyclophosphamide, and lomustine (MACC) had been unsuccessful. PMID- 2553238 TI - High-dose cytosine arabinoside in relapsed and refractory non-Hodgkin's lymphoma. Limited role as a single agent. AB - Cytosine arabinoside (ara-C) is an S-phase active antineoplastic agent used in the treatment of several hematologic malignancies. We did a retrospective analysis on 48 patients with relapsed or refractory non-Hodgkin's lymphoma (NHL) who were treated with high-dose ara-C between 1982 and 1987. All patients received between 9 and 24 g/m2/course of treatment with 90% receiving 12g/m2/treatment. Fifteen patients (25%) achieved a complete or partial remission. Of these, five (10%) were complete responders. Long-term disease-free survival was maintained only in those responders who were consolidated with autologous bone marrow transplantation. Toxicity was primarily myelosuppression. Seventy-five percent of patients developed fever and neutropenia with an equivalent percent having severe thrombocytopenia sufficient to require platelet transfusion. Five patients (10%) had significant neurotoxicity. There were seven treatment-related deaths (15%). Patients in whom tumors contained a small cell component responded better than patients with purely large cell or undifferentiated lymphomas (50% versus 15%, P = 0.024). We conclude that high dose ara-C given as 3g/m2 every 12 hours for four doses has a limited role when used as a single agent in the treatment of relapsed or refractory non-Hodgkin's lymphoma. However, there are subsets of patients who may respond favorably to this therapy. PMID- 2553239 TI - Human papillomaviruses and the pathogenesis of cervical neoplasia. A study by in situ hybridization. AB - In a previous topographic study of cervical conization specimens, condylomatous changes were commonly present in direct contact with intraepithelial neoplasia and were always located distal (ectocervical side) to the neoplasia. Viral DNA was detected by in situ hybridization using 35S-labeled nick translated DNA probes in 50 of 70 cases (71%) which had adequate lesions: HPV-16 in 30, HPV-18 in ten, HPV-31 in six, and multiple types in four cases. HPV-6/11 was detected only once, in a multiple infection. As a rule, a positive cervix contained a single virus type, and the same virus type was found in condylomatous and neoplastic areas. The results suggest that the neoplastic process is initiated in the area of condyloma toward the endocervix and, once established, extends proximally toward the cervical canal. Capsid antigen was detected in 19 cases, indicating that a proportion of the high-grade lesions is potentially infectious. PMID- 2553240 TI - Mixed germ cell-sex cord stroma tumor of the testis. A report with ultrastructural findings. AB - The case of a 41-year-old man with testicular mixed germ cell-sex cord stroma tumor is reported. The patient noticed a gradual, painless, right-sided testicular enlargement 4 years before being treated by radical orchiectomy. The testis contained a centrally located tumor. There was no evidence of metastases. An histologic examination and an ultrastructural study showed that the tumor was composed of germ cells and sex cord derivatives; the typical features of mixed germ cell-sex cord stroma tumor were present. The patient is well and disease free 2 years after the operation. Currently, none of the testicular tumors of this type has been associated with metastases or was overgrown by malignant germ cell tumors, and radical orchiectomy resulted in complete cure. The literature regarding this entity is reviewed, the differential diagnosis is discussed, and the importance of making the correct diagnosis is emphasized. PMID- 2553242 TI - Clinical experience in 20 hepatic resections for peripheral cholangiocarcinoma. AB - During the 10-year period from 1978 to 1987, hepatic resections were performed on 20 patients with peripheral cholangiocarcinoma (PCC). Nine of these patients were men and 11 were women (mean age, 48.5 years). Among them, 80% had intrahepatic stones with recurrent cholangitis. The 20 patients were subdivided into the following three groups: Group I (12 patients with surgery for PCC); Group II (4 patients with surgery for chronic cholangitis [but the final pathologic diagnosis confirmed PCC]); and Group III (4 patients with surgery for space-occupying liver lesions). No early postoperative mortality was noticed. The few complications that occurred were related to surgery for hepatolithiasis. Postoperative wound infection was the most common complication. The overall mean survival time was 20.5 months. Four patients survived for more than 3 years; one was even alive for more than 5 years after surgery. PMID- 2553241 TI - Predicting the rate and extent of locoregional failure after breast conservation therapy for early breast cancer. AB - Five hundred thirty-six cases of early breast cancer (T0 to T2, N0 to N1) treated with breast conservation therapy were monitored for a median interval of 64 months. Fifty-five locoregional failures occurred after a median interval of 40 months; the actuarial failure rate was 9% at 5 years and 19% at 10 years. Factors predicting locoregional failure were sought. Young patients had a higher risk of failure than older patients, although their actuarial survival rates were not different. Locoregional failure was defined as advanced if tumor involved skin or was fixed to the chest wall, the diameter was greater than 5 cm, or unresectable nodes were present. There was a significantly higher incidence of advanced recurrence with higher initial tumor stage. Overall, the 5-year survival rate after treatment of locoregional recurrence was 63%. Advanced locoregional failure, however, resulted in a median survival time of 37 months. Further study is required to explain the increased failure rate in younger women. Advanced locoregional failure rarely occurred after treatment of Stage 0 or Stage I tumors, supporting the selection of patients with early disease for breast conservation therapy. PMID- 2553244 TI - Malignant fibrous histiocytoma (giant cell type) of the pancreas. A distinctive variant of osteoclast-type giant cell tumor of the pancreas. AB - Malignant giant cell tumors of the pancreas are rare neoplasms which have been generally thought to represent epithelial malignancies of either acinar or ductal epithelium. The authors have studied a tumor of the pancreas that was characterized histologically by a proliferation of benign-appearing osteoclast type giant cells in association with atypical, often bizarre mononuclear cells. Immunohistochemical studies demonstrated negative staining of the tumor cells with epithelial markers, including low-molecular weight keratins, carcinoembryonic antigen and epithelial membrane antigen, and positive staining with vimentin antibodies, supporting a fibroblastic line of differentiation. Electron microscopic examination also showed absence of ultrastructural features of epithelial differentiation such as microvilli, intercellular junctions, or desmosomes. The authors believe the current case represents a true sarcoma of the pancreas, currently best classified as a malignant fibrous histiocytoma, giant cell type. This tumor should be distinguished from the epithelial type of osteoclastic giant cell tumor of the pancreas. PMID- 2553243 TI - A cell kinetic study of pulmonary adenocarcinoma by an immunoperoxidase procedure after bromodeoxyuridine labeling. AB - By in vitro labeling with bromodeoxyuridine (BrdU) and immunohistochemical staining with anti-BrdU monoclonal antibody, the tumor cell kinetics were investigated in 52 resected pulmonary adenocarcinomas. The values of the BrdU labeling index (LI) varied widely among cases of adenocarcinoma in comparison with four cases of small cell carcinoma and ten cases of squamous cell carcinoma examined as controls. The LI was below 3.0 in 60% of the adenocarcinomas. When the LI was analyzed according to the cytologic subtypes of adenocarcinoma, the values were low for goblet cell type and type II alveolar cell type, whereas the values for the bronchial surface epithelial cell type varied widely (average, 5.2) and those for the Clara cell type were intermediate (average, 1.8). A comparison of the LI with TNM classification, histologic differentiation, degree of nuclear atypia, and mitotic cell count showed a correlation between the LI and each of these prognostic factors. Thus, the possibility of the BrdU LI being a useful new prognostic factor of pulmonary adenocarcinomas was suggested. PMID- 2553245 TI - Maternal transmission of hepatitis B virus in childhood hepatocellular carcinoma. AB - Fifty-one children, aged between 3 and 16 years, were diagnosed to have hepatocellular carcinoma (HCC) in the last 15 years. Serum hepatitis B surface antigen (HBsAg) was positive in all the latter 34 HCC children checked by radioimmunoassay, and was positive in five of the 17 earlier HCC children studied by double immunodiffusion method. Serum HBsAg was studied in 31 mothers of the latter 33 HCC children, including two pairs of affected siblings and was positive in 29 (94%). The positive rate was much higher than that (50%) of the mothers of control HBsAg carrier children (P less than 0.001). The serum HBsAg positive rate was also higher in the siblings of HCC children than that of the control group. On the contrary, the serum HBsAg positive rate was not different between the fathers of HCC children and that of the control group. Our observation demonstrated that transmission of hepatitis B virus from the mothers during the perinatal period or early childhood is the most important mode of hepatitis B virus infection in HCC children in Taiwan. PMID- 2553246 TI - CT manifestations of a ruptured hepatic tumor after transcatheter arterial embolization. AB - There are few reports of the radiologic diagnosis of ruptured hepatic tumors. In a patient with right upper abdominal pain and impending shock, angiography demonstrated a hypervascular hepatic tumor, and CT imaged an extrahepatic mass suggestive of a hematoma. Following transcatheter arterial embolization with Lipiodol Ultrafluide and gelatin-sponge, multiple contiguous CT sections revealed numerous lipiodol droplets adjacent to a lipiodol-containing hepatic tumor, clearly outside the liver. These findings were indicative of a ruptured hepatic tumor. After embolization, the patient's condition improved and he was discharged. PMID- 2553247 TI - Influence of vitamin B2 on formation of chromium(V), alkali-labile sites, and lethality of sodium chromate(VI) in Chinese hamster V-79 cells. AB - The effect of vitamin B2 on the cellular reduction and cytotoxicity of chromate(VI) was studied using Chinese hamster V-79 cells. Electron spin resonance studies showed that incubation of cells with Na2CrO4 resulted in the formation of both chromium(V) and chromium(III) complex and that cellular pretreatment with riboflavin (Vitamin B2) for 24 h prior to exposure increased the level of chromium(V) complex, but the level of chromium(III) remained unchanged. Analysis of flavin derivatives revealed that pretreatment with vitamin B2 increased free riboflavin without altering flavin adenine dinucleotide and flavin mononucleotide. In addition, the level of the flavoenzyme glutathione reductase, which is capable of reducing chromate, was unaffected by pretreatment with vitamin B2. However, treatment of cells with vitamin B2 and Na2CrO4 augmented the inhibition of glutathione reductase attributable to Na2CrO4 alone. Using a colony-forming assay, pretreatment with vitamin B2 resulted in a decrease of cytotoxicity after exposure to the lethal concentration of chromate (15 microM) but did not affect the cytotoxicity at sublethal concentration of this metal (5-7.5 microM). Alkaline elution studies demonstrated that Na2CrO4 induced alkali-labile sites in the DNA of cells in a concentration-dependent manner (5-15 microM) and pretreatment with vitamin B2 resulted in an increase of these DNA lesions at all concentrations of Na2CrO4. The results, showing that vitamin B2 enhances chromate-induced alkali-labile lesions and chromium inhibition of glutathione reductase, might be due to an increase of chromium(V) species, possibly through its ability to directly reduce chromium(VI). The results also suggest that the extent of DNA lesions induced by chromate may not correlate directly with the cytotoxic effects of this metal. PMID- 2553248 TI - Altered formation of DNA in human cells treated with inhibitors of DNA topoisomerase II (etoposide and teniposide). AB - We have investigated the importance of DNA topoisomerase II for the formation of mammalian DNA replication intermediates. Treatment with the DNA topoisomerase II inhibitor etoposide (teniposide) prevents the formation of large intermediates, such as 10-kilobase DNA, but allows the formation of small intermediates, i.e., Okazaki fragments. In untreated cells, there is a distinct stage in which the 10 kilobase DNA intermediates are joined before the appearance of mature chromatin. We find that pretreatment with etoposide (teniposide) prevents the appearance of this stage. When the protocol is reversed and the cells contain labeled 10 kilobase DNA before exposure to the drugs, one can detect the stage. PMID- 2553249 TI - Potentiation of interferon induction of class I major histocompatibility complex antigen expression by human tumor necrosis factor in small cell lung cancer cell lines. AB - The response of class I major histocompatibility complex antigen expression to in vitro administration of interferon and tumor necrosis factor alpha (TNF-alpha) was measured using class I major histocompatibility complex-deficient small cell lung cancer cell lines. Significant induction also was observed using gamma interferon (IFN-gamma) alone, whereas TNF-alpha alone yielded only modest induction. Classic small cell lung cancer cell lines NCI-H146 and NCI-H209 best demonstrated synergistic HLA and beta 2-microglobulin antigen induction with IFN gamma and TNF-alpha with the following dose schedule: 3-6 days of TNF-alpha (200 units/ml) followed by 48 h of IFN-gamma (100 IU/ml). Induction was quantitated using an 125I-Protein A radioimmunoassay. Synergistic induction of the HLA and beta 2-microglobulin surface antigens on NCI-H146 was also possible with alpha interferon and TNF-alpha but required a higher concentration of the interferon, i.e., 3-6 days of TNF-alpha (200 units/ml) followed by 48 h of alpha interferon (1000 units/ml). Small cell lung cancer cell line NCI-H146 was further studied for expression of major histocompatibility complex messenger RNA using the optimal doses and sequence of addition of IFN-gamma and TNF-alpha as indicated above. A significant induction with IFN-gamma alone and synergistic induction with both IFN-gamma and TNF-alpha was quantitated for both HLA-A2 and beta 2 microglobulin transcripts using Northern blot analysis. Incubation with relatively low subcytotoxic doses of IFN-gamma and TNF-alpha also resulted in a marked synergistic decrease in c-myc message. PMID- 2553250 TI - Growth inhibition of human breast cancer cells in vitro with an antibody against the type I somatomedin receptor. AB - Insulin and insulin-like growth factors (IGFs) stimulate the growth of human breast cancer cells in vitro. The type I somatomedin receptor (SR), expressed in these cells, may mediate the mitogenic effects of these peptides. We have examined the effect of type I SR blockade on human breast cancer growth with a monoclonal antibody (alpha-IR3) that blocks the receptor binding domain. alpha IR3 inhibited binding of 125I-IGF-I in all breast cancer cell lines tested. Binding affinity of alpha-IR3 was 2 to 5 times higher than that of IGF-I in MDA 231 (Kd 2.1 nM) and MCF-7 cells (Kd 0.6 nM), respectively. In the presence of 10% calf serum, the antibody inhibited anchorage-independent growth of six of seven breast cancer cell lines. This inhibition was reversible with excess IGF-I. In serum-free medium, alpha-IR3 blocked IGF-I-stimulated DNA synthesis in four of four breast cancer cell lines (MCF-7, ZR75-1, MDA-231, and HS578T). However, the antibody did not inhibit basal growth of any of the breast cancer cell lines in serum-free conditions. In three estrogen receptor-positive, estrogen-responsive breast cancer cell lines (MCF-7, ZR75-1, and T47D), type I SR blockade with alpha IR3 failed to block estrogen-stimulated DNA synthesis or cell proliferation, indicating that secreted IGF activity is not the sole mediator of the growth effects of estrogen. In conclusion, antibody-mediated type I SR blockade does not inhibit basal growth of breast cancer cells under serum-free conditions, arguing against a critical autocrine role of endogenously secreted IGF activity in vitro. However, type I SR blockade inhibits breast cancer cell growth in the presence of serum, suggesting that serum IGFs might be critical endocrine or paracrine regulators of human breast cancer. PMID- 2553251 TI - Altered adrenergic response and specificity of the receptors in rat ascites hepatoma AH130. AB - Adenylate cyclase activation through adrenergic receptors in rat ascites hepatoma (AH) 130 cells in response to adrenergic drugs was studied, and receptor binding and displacement were compared with those of normal rat hepatocytes. Epinephrine (Epi) and norepinephrine (NE) activated AH130 adenylate cyclase about half as much as isoproterenol (IPN) but equaled IPN after treatment with the alpha antagonist phentolamine or islet-activating protein (IAP). The three catecholamines in hepatocytes were similar regardless of phentolamine or IAP. These catecholamines activated adenylate cyclase in order of IPN greater than NE greater than Epi in AH130 cells but IPN greater than Epi greater than NE in hepatocytes. We then used the alpha 1-selective ligand [3H]prazosin, the alpha 2 selective ligand [3H]clonidine, and the beta-ligand [125I]iodocyanopindolol [( 125I]ICYP), and found that AH130 cells had few prazosin-binding sites, about eight times as many clonidine-binding sites with high affinity, and many more ICYP-binding sites than in hepatocytes. The dissociation constant (Ki) of the beta 1-selective drug metoprolol by Hofstee plots for AH130 cells was lower than that for hepatocytes. The inhibition of specific ICYP binding by the beta 2 selective agonist salbutamol for AH130 cells gave only one Ki value which was much higher than both high and low Ki values of the drug for hepatocytes. These findings indicate that the alpha- and beta-adrenergic receptors in hepatocytes are predominantly alpha 1-type and beta 2-type, but that those in AH130 cells are predominantly alpha 2-type and beta 1-type, and the low adrenergic response of AH130 cells is due to the dominant appearance of alpha 2-adrenergic receptors, linked with the inhibitory guanine-nucleotide binding regulatory protein, instead of alpha 1-adrenergic receptors, and beta 1-adrenergic receptors with low affinity for the hormone. PMID- 2553252 TI - Production of and responsiveness to transforming growth factor-beta in normal and oncogene-transformed human mammary epithelial cells. AB - Transforming growth factors-beta (TGF beta) are a family of closely related, ubiquitously expressed growth factors with the common properties of induction of growth inhibition and expression of differentiation-related markers in epithelial cells. We investigated the role of TGF beta 1 in growth regulation of normal human mammary epithelial cells and in benzo(a)pyrene immortalized sublines further transformed by oncogenes in retroviral vectors. The normal cells were markedly growth inhibited by TGF beta 1, produced TGF beta in a latent form, and expressed TGF beta receptors. In the immortalized cells, both TGF beta-induced growth inhibition and TGF beta receptor binding were reduced. With the single oncogenes v-Ha-ras, v-mos, and SV40 T, growth sensitivity to TGF beta 1 increased, but TGF beta production or TGF beta receptor expression was not altered. Transformation to full malignancy by both SV40 T and v-Ha-ras led to escape from growth inhibition by TGF beta under anchorage-independent, but not anchorage-dependent, conditions without affecting TGF beta production or receptor characteristics. Thus, modulation of TGF beta growth responsiveness in these normal and oncogene transformed human mammary epithelial cells apparently occurs at a level distal to TGF beta receptor binding and is not solely correlated to expression of transforming oncogenes. Further, modulation of TGF beta production is not an indicator of malignant transformation in this system. PMID- 2553253 TI - Expression of human DNA topoisomerase I in yeast cells lacking yeast DNA topoisomerase I: restoration of sensitivity of the cells to the antitumor drug camptothecin. AB - Yeast Saccharomyces cerevisiae strains that are permeable to the antitumor alkaloid camptothecin are killed by the drug if they express DNA topoisomerase I, the cellular target of the drug (J. Nitiss and J.C. Wang, Proc. Natl. Acad. Sci. USA, 85: 7501-7505, 1988). We show that in a yeast strain permeable to camptothecin but lacking DNA topoisomerase I, sensitivity to the drug was restored upon expression of human DNA topoisomerase I from a plasmid-borne human complementary DNA clone. When the human enzyme was expressed from a galactose inducible, glucose-repressible yeast promoter, PGAL1, sensitivity to camptothecin was observed in the presence of galactose but not in the presence of glucose. Expression of human DNA topoisomerase I in Escherichia coli was also demonstrated by the complementation of a conditional lethal E. coli DNA topoisomerase I mutant. These systems can be used in the study of human DNA topoisomerase I camptothecin interactions and in the screening of additional therapeutics targeting the enzyme. PMID- 2553254 TI - Differential requirement of DNA replication for the cytotoxicity of DNA topoisomerase I and II inhibitors in Chinese hamster DC3F cells. AB - The cytotoxicity of topoisomerase inhibitors is thought to result from the induction of enzyme-mediated DNA breaks. The fact that these breaks reverse rapidly in cells programmed to die, led us to investigate further the cytotoxic mechanisms of topoisomerase I (camptothecin) and topoisomerase II inhibitors (VP 16 and amsacrine) in Chinese Hamster lung fibroblasts (DC3F). Exposures (30 min) to camptothecin produced limited cell killing with approximately 20% of the cells naturally resistant. This resistance was overcome by increasing the drug exposure time. Inhibition of DNA synthesis by 5-min pretreatments with aphidicolin or hydroxyurea abolished the cytotoxicity of camptothecin without changing the level of camptothecin-induced DNA breaks. A good correlation was found between the degree of DNA synthesis inhibition by aphidicolin and the reduction of camptothecin cytotoxicity. In similar experiments performed with topoisomerase II inhibitors, aphidicolin prevented only partially against VP-16- and amsacrine induced cytotoxicities, yet had no effect upon drug-induced DNA breaks. These results indicate that the production of topoisomerase-mediated DNA breaks by antitumor drugs is not sufficient for cell killing. Instead, an interference of moving DNA replication forks with drug-stabilized topoisomerase-DNA complexes is critical for cell death. The cytotoxicity of camptothecin seemed to be completely related to this process, while that of topoisomerase II inhibitors seemed to involve additional mechanisms in DC3F cells. PMID- 2553256 TI - Evaluation of the cocarcinogenic effect of wounding in Rous sarcoma virus tumorigenesis. AB - Chickens given injections of Rous sarcoma virus form sarcomas at the site of inoculation (primary tumor) and at the site of experimentally introduced wounds (wound tumor). This latter finding provides a model system to study systematically the mechanisms underlying the cocarcinogenic effects of wounding. Our experiments show the following. (a) Chickens inoculated with a Rous sarcoma virus-derived, replication-defective virus construct fail to elaborate wound tumors in spite of aggressively growing primary tumors. We thus rule out metastasis as a mechanism and conclude that infectious virus is required for wound tumor formation; (b) using bromodeoxyuridine incorporation and immunofluorescence on frozen sections we demonstrate proliferation in the unwounded wing in cell types which are normally targets for Rous sarcoma virus infection and transformation and conclude that proliferation per se is not sufficient to induce wound tumors; (c) using immunohistochemistry for the viral protein p19gag we show that wounding induces virus expression in fibroblasts of newly forming granulation tissue 2 days after injury. We also demonstrate expression of viral mRNA in wound tumors by in situ hybridization with a v-src probe. We discuss the possibility of activation of integrated, silent virus or the preferential infection of a special target cell population as a result of wounding as well as the potential role of wound factors in transformation. PMID- 2553257 TI - Clinical management of familial adenomatous polyposis. AB - Familial adenomatous polyposis is a generalized growth disorder. It manifests itself in a catastrophic way with the inevitable development of colorectal cancer if left untreated. The aim of clinical management should be to detect this disease at its earliest possible stage by treatment of the family as a unit and identification of those at risk with appropriate screening. Early surgical intervention with, most commonly, colectomy with ileorectal anastomosis or, in more advanced cases, with colectomy, rectal mucosectomy and ileoanal pouch procedure is appropriate. Following colorectal cancer the major risks of death from this disease include upper gastrointestinal cancer, desmoid tumour and a number of other malignancies throughout the body. The exact magnitude of the risk of malignant degeneration of these extracolonic manifestations is as yet uncertain. Surveillance is required particularly for upper gastrointestinal adenomas, as there is a significant risk for the development of duodenal carcinoma and it is obvious that prophylactic colectomy alone does not cure this disease predictably. The role of a familial polyposis registry in managing these patients is important not only in maintaining compliance with surveillance and therefore early detection of the disease but also in educating the family members and gaining long-term follow-up data on these cases to more accurately define the risk of death from extracolonic malignancy. PMID- 2553255 TI - Yttrium-90 and iodine-131 radioimmunoglobulin therapy of an experimental human hepatoma. AB - Therapeutic trials were performed on the HepG2 human hepatoblastoma implanted s.c. in the athymic nude mouse. Animals were treated with polyclonal and monoclonal antiferritin and control antibodies labeled with either iodine-131 (131I) or yttrium-90 (90Y). Administration of 400 muCi of 131I-labeled polyclonal antiferritin or 300 muCi of 90Y-labeled polyclonal antiferritin significantly increased survival (P less than 0.001). There were no tumor cures with radiolabeled polyclonal antibody therapy. Animals treated with 200 or 300 muCi of 131I-labeled monoclonal antiferritin (QCI054) did not show increased survival compared to controls. Although 400 muCi of 131I-labeled QCI significantly prolonged survival, treatment resulted in no long-term survivors. Monoclonal antiferritin labeled with 90Y significantly prolonged survival of animals (P less than 0.001) at doses of 100, 200, or 300 muCi compared with untreated controls. Fifty % of the animals treated with 200 muCi and 75% of the animals treated with 300 muCi showed no evidence of disease at 140 days following treatment. Four hundred muCi of 90Y-labeled QCI proved toxic to the animals. Increased survival was accompanied by a decrease in tumor mitotic rate and increase in cellular polymorphism as determined by pathological examination. The radiation dose absorbed in the tumor correlated directly with tumor response following treatment. The absorbed dose in tumors for complete decay of the isotope ranged from 165 and 330 cGy at the periphery and center of small tumors for an administered activity of 200 muCi of 131I-labeled polyclonal antiferritin, to 7,573 and 12,400 cGy for 300 muCi of 90Y-labeled monoclonal antiferritin QCI. PMID- 2553258 TI - Chronic diazepam treatment induces an increase in peripheral benzodiazepine binding sites. AB - The effect of chronic (21 days) diazepam treatment (0.5 mg/kg, intraperitoneally) on [3H]PK 11195 binding to the heart and cerebral cortex was studied. A significant increase in the density of peripheral benzodiazepine binding sites in the heart (18%) and cerebral cortex (19%) was observed. Following five days of withdrawal, the maximal binding capacity of these sites returned to normal control values. No alteration in the equilibrium dissociation constant was observed during the treatment and withdrawal period. PMID- 2553259 TI - Effect of selected antiarrhythmic drugs on the superoxide anion production by polymorphonuclear neutrophils in vitro. AB - The oxygen radicals produced by polymorphonuclear neutrophils (PMN) during ischemia and reperfusion play an important role in the development of arrhythmias. We investigated the influence of selected antiarrhythmic drugs on PMN superoxide anion production. Amiodarone, verapamil and propranolol significantly decreased the stimulated superoxide anion production, which may be an additional mechanism of their antiarrhythmic activity. The fast sodium channel inhibitors lidocaine, procainamide and mexiletine affected neither spontaneous nor stimulated superoxide anion production. PMID- 2553260 TI - The effects of enalapril on blood pressure, renal hemodynamics, and renal function. AB - Since the introduction of angiotensin converting enzyme (ACE) inhibitors into clinical use, much information has been accumulated in animal models and man regarding their effects on renal function in different disease states. Enalapril, the first nonsulfhydryl ACE inhibitor approved for general use in the United States, has demonstrated efficacy and safety in controlling blood pressure in patients with essential hypertension, renal parenchymal disease, renovascular hypertension, and diabetes with hypertension. Enalapril also appears capable of attenuating the progressive nature of renal disease in experimental models of chronic renal failure and diabetic nephropathy, perhaps through lowering intraglomerular pressures. The excellent blood pressure-lowering effects of ACE inhibitors, coupled with their potential to ameliorate renal hemodynamic abnormalities, make these compounds attractive for use in these clinical states. PMID- 2553261 TI - Analysis of human neutrophil granule protein composition in chronic myeloid leukaemia by immuno-electron microscopy. AB - In this study immuno-electron microscopy was used to assay, semi-quantitatively, the granule contents of elastase, lactoferrin, lysozyme and myeloperoxidase in human peripheral blood neutrophils from 13 chronic myeloid leukaemia patients in the chronic phase of the disease and from normal non-smoking donors. The fixation conditions that adequately preserved the antibody binding capacities of these antigens and reasonably preserved the ultrastructure of the neutrophils were selected by light-microscopic immunoperoxidase cytochemistry on cytospin smears. Immunogold cytochemistry on LR White resin sections localised elastase and myeloperoxidase to the primary granules, lactoferrin to the secondary granules and lysozyme to both types of granule. When applicable, peroxidase cytochemistry was combined with immunogold staining making it easier to distinguish the primary from the secondary granules. A comparison of the immunolabelling density values obtained for the leukaemic and normal states revealed no significant abnormalities in the immunoreactivity patterns for any of these neutrophil granule antigens in the leukaemic patients. All 13 patients gave normal immunostaining reactivities for these neutrophil granule proteins. Consequently the distribution patterns of these proteins, as shown in this study, cannot be used as indices in distinguishing chronic myeloid leukaemic neutrophils from normal neutrophils. PMID- 2553262 TI - Distribution of 5'-nucleotidase in the renal interstitium of the rat. AB - The hydrolysis of 5'-AMP by 5'-nucleotidase is the main source of adenosine. In various tissues adenosine is a local mediator adjusting the organ work to the available energy. In the kidney it regulates renal hemodynamics, glomerular filtration rate and renin release via specific receptors of the arteriolar walls. By immunocytochemistry we identified interstitial and tubular sites of 5' nucleotidase in the rat kidney. In the interstitium the enzyme was detected only in the cortical labyrinth, the compartment that comprises all arteriolar vessels besides other putative targets of adenosine. The 5'-nucleotidase-positive cells of the interstitium were identified as fibroblasts. The fibroblasts are in close contact with the tubules as well as with the vessels. Thus, any 5'-AMP released by the tubules into the interstitial space would be converted to adenosine in the direct vicinity of its assumed targets. Adenosine produced by tubular cells would hardly have access to its known targets, since 5'-nucleotidase is restricted to the luminal cell surface. Pathological events affecting the fibroblasts might influence renal function by modifying the interstitial adenosine production. PMID- 2553263 TI - Stereological and functional investigations on isolated adrenocortical cells: zona fasciculata/reticularis cells of chronically ACTH-treated rats. AB - The morphology and function of isolated inner (zona fasciculata/reticularis) adrenocortical cells of rats pretreated with ACTH for 3, 6, 9 or 12 days were investigated. ACTH treatment induced a notable time-dependent enhancement in the steroidogenic capacity (corticosterone production) and growth of inner cells. The volumes of cells, mitochondrial compartment, membrane space [the cellular space occupied by smooth endoplasmic reticulum (SER) membranes] and lipid-droplet compartment, as well as the surface area of mitochondrial cristae and SER tubules, were increased in relation to the duration of ACTH pretreatment, and showed a highly significant positive linear correlation with both basal and stimulated corticosterone production. The acute exposure of isolated cells to ACTH provoked a striking lipid-droplet depletion, the extent of which was linearly and positively correlated with stimulated corticosterone secretion. The hypertrophy of the mitochondrial compartment and SER are interpreted as the morphological counterpart of the enhanced steroidogenic capacity of inner adrenocortical cells, inasmuch as the enzymes of steroid synthesis are located in these two organelles, and it is well known that chronic ACTH exposure stimulates the de novo synthesis of many of them in vivo. The rise in the number of lipid droplets, in which cholesterol is stored, is interpreted as being due to the fact that, under chronic ACTH treatment, the processes leading to cholesterol accumulation in adrenocortical cells (exogenous uptake and endogenous synthesis) exceed those of its utilization in basal steroid secretion. Cholesterol accumulated in lipid droplets as a reserve material may be rapidly utilized after acute ACTH exposure to meet the needs of the enhanced steroidogenic capacity of adrenocortical cells. PMID- 2553264 TI - Effects of corticotrophin-releasing hormone on corticotrophs in anterior pituitary gland allografts in hypophysectomized, orchidectomized hamsters. AB - We investigated the effects of corticotrophin-releasing hormone (CRH) on the percentage of anterior pituitary gland (APG) cells which are corticotrophs as well as the size and shape of corticotrophs. Pituitary glands were removed from 7 week-old male hamsters and placed beneath the renal capsules of hamsters that had been hypophysectomized and orchidectomized 3 weeks previously. Beginning 6 days after each host had received a single allograft, each was injected subcutaneously twice daily with 4 micrograms CRH or vehicle for 16 days. Six hosts in each group were decapitated 16 h after the last injection. Sections of anterior pituitary tissue were stained for ACTH and with hematoxylin. The percentage of corticotrophs among APG cells was greater in allografts exposed to exogenous CRH (approximately 20%) than in allografts exposed to vehicle (approximately 15%). Exposure to exogenous CRH increased the cross-sectional area of corticotroph cells in allografts to values greater than those measured for corticotrophs in allografts exposed to vehicle, without altering the shape of cells. Results of subsequent studies suggested that hamsters with allografts injected with vehicle do not release ACTH and that exogenous CRH causes an abrupt release of ACTH from allografts. These results indicate that CRH releases ACTH from ectopic corticotrophs and that administration of CRH can increase corticotroph size and the percentage of APG cells that are corticotrophs. PMID- 2553265 TI - Signal transmission by the insulin-like growth factors. PMID- 2553266 TI - HIV-1 Tat protein increases transcriptional initiation and stabilizes elongation. AB - We studied regulation of human immunodeficiency virus-1 (HIV-1) transcription by Tat and, for comparative purposes, by the adenovirus E1A protein. These two trans activators exerted different effects. Two classes of HIV-1-promoted cytoplasmic RNA were detected, one class corresponding to full-length transcripts and the other to transcripts ending 55 and 59 nucleotides from the transcription start. Tat increased the level of the full-length class only, whereas E1A increased the levels of both classes of RNA. We also measured the effects of Tat and E1A on RNA synthesis rates. Without trans-activators, HIV-1-directed transcription was relatively weak and exhibited a marked polarity. Both Tat and E1A dramatically increased promoter-proximal transcription, while only Tat suppressed transcriptional polarity. Mutations in the TAR element did not influence basal transcription rates or the response to E1A, but eliminated trans-activation by Tat. We propose that Tat acts through TAR to increase initiation complex formation on the HIV-1 promoter and to stabilize complexes during elongation. PMID- 2553267 TI - The ski oncogene induces muscle differentiation in quail embryo cells. AB - Quail embryo cells (QECs) are primary cultures of fibroblastoid cells that become myogenic after infection with avian retroviruses expressing the ski oncogene (SKVs). ski also stimulates proliferation of QECs and induces morphological transformation and anchorage-independent growth. Paradoxically, ski-transformed clones picked from soft agar are capable of muscle differentiation. ski-induced differentiation is essentially indistinguishable from that of uninfected myoblasts in culture with regard to muscle-specific gene expression, commitment, and inhibition by growth factors or other oncogenes. However, ski-induced myoblasts have less stringent requirements for growth and differentiation. Uninfected QECs cannot differentiate and do not express an early marker for the myogenic lineage. Clonal analysis indicates that at least 40% of QECs are converted by ski to differentiating myoblasts. The data suggest that ski induces either the capacity for differentiation in an "incompetent" muscle precursor or the determination of nonmyogenic cells to the myogenic lineage. PMID- 2553268 TI - Drosophila P element transposase recognizes internal P element DNA sequences. AB - Drosophila P transposable elements encode an 87 kd trans-acting protein, transposase, that is required to catalyze P element transposition and excision. We show here that purified transposase is a site-specific DNA binding protein. P element transposase does not interact with the terminal 31 bp inverted repeats but instead interacts specifically with an internal 10 bp consensus sequence present at both the 5' and 3' ends of P element DNA. These binding sites lie within sequences known to be important for transposition in vivo. Transposase also displays an unusually high nonspecific affinity for DNA. The transposase binding site at the 5' and overlaps sequences we show to be essential for transcription from the P element promoter in vitro, which raises the possibility that either transposase or the related 66 kd P element protein may affect P element transcription. From these and other observations, we suggest that the P element transposition reaction probably requires the binding of additional Drosophila protein factors to the terminal DNA sequences. PMID- 2553269 TI - Intramolecular transposition by Tn10. AB - Transposon Tn10 promotes the formation of a circular product containing only transposon sequences. We show that these circles result from an intramolecular transposition reaction in which all of the strand cleavage and ligation events have occurred but newly created transposon/target junctions have not undergone repair. The unligated strand termini at these junctions are those expected according to a simple model in which the target DNA is cleaved by a pair of staggered nicks 9 bp apart, transposon sequences are separated from flanking donor DNA by cleavage at the terminal nucleotides on both strands (at both ends) of the element, and 3' transposon strand ends are ligated to 5' target strand ends. The stability of the unligated junctions suggests that they are protected from cellular processing by transposase and/or host proteins. We propose that the nonreplicative nature of Tn10 transposition is determined by the efficiency with which the nontransferred transposon strand is separated from flanking donor DNA and by the nature of the protein-DNA complexes present at the strand transfer junctions. PMID- 2553270 TI - A specific class of IS10 transposase mutants are blocked for target site interactions and promote formation of an excised transposon fragment. AB - We report the identification and characterization of a class of IS10 transposase mutants that carry out only some of the steps required for transposition. These mutants were identified among transposition-defective mutants as a specific subclass that retains the wild-type ability to induce SOS functions in the presence of transposon ends. Mutants of this class successfully promote excision of the element from its donor site, but do not promote transfer of the transposon sequences to a target site. SOS induction presumably results from the degradation of the donor site. Uniquely among transposition-defective mutants, SOS+ Tnsp- mutants promote the formation of a new product, the excised transposon fragment (ETF), which consists of the transposon excised from the original donor molecule by double-strand breaks at the transposon ends. SOS+ Tnsp- mutants identified thus far define two patches of amino acids that might correspond to regions of different function. A single additional mutation maps within a region that is highly conserved among IS element transposases. The existence of SOS+ Tnsp- mutants and the structure of the ETF provide strong support for the previously proposed nonreplicative model of Tn10/IS10 transposition. PMID- 2553271 TI - Neural and developmental actions of lithium: a unifying hypothesis. PMID- 2553272 TI - Self-catalyzed linkage of poliovirus terminal protein VPg to poliovirus RNA. AB - The poliovirus terminal protein, VPg, was covalently linked to poliovirus RNA in a reaction that required synthetic VPg, Mg2+, and a replication intermediate synthesized in vitro. The VPg linkage reaction did not require the viral polymerase, host factor, or ribonucleoside triphosphates and was specific for template-linked minus-strand RNA synthesized on poliovirion RNA. The covalent nature of the bond between VPg and the RNA was demonstrated by the isolation of VPg-pUp from VPg-linked RNA. A model is proposed in which the tyrosine residue in VPg forms a phosphodiester bond with the 5'UMP in minus-strand RNA in a self catalyzed transesterification reaction. It appears that either the RNA, VPg, or a combination of both forms the catalytic center for this reaction. PMID- 2553273 TI - Transcriptional activator nuclear factor I stimulates the replication of SV40 minichromosomes in vivo and in vitro. AB - SV40 DNA replication in vivo is greatly stimulated by cis-acting transcriptional elements. We studied a model viral chromosome containing a single binding site for the cellular transcriptional activator, nuclear factor I (NF-I/CTF), located adjacent to the replication origin. The presence of the NF-I recognition site increased replication efficiency over 20-fold in vivo. Purified NF-I had little effect on the replication efficiency in the standard SV40 cell-free system when the template was introduced as naked DNA. However, NF-I specifically prevented the repression of DNA replication that occurred when the template was preassembled into chromatin. Our data support a model in which the binding of a transcriptional activator perturbs the local distribution of nucleosomes, thereby increasing the accessibility of the origin region. PMID- 2553274 TI - T cell receptor gamma gene status of human alpha/beta+ and gamma/delta+ T cell clones: absence of V9JP rearrangements in alpha/beta+ clones is not a result of a lack of rearrangements involving more 5' J gamma segments. AB - T cell receptor (TCR) gamma gene rearrangements were examined in panels of human T cell clones expressing TCR alpha/beta or gamma/delta heterodimers. Over half of the alpha/beta+ clones had both chromosomes rearranged to C gamma 2 but this was the case for only 20% of the gamma/delta+ clones. While more than half of the gamma/delta+ clones showed a V9JP rearrangement, this configuration was absent from all 49 alpha/beta+ clones analysed. However, this was not a result of all rearrangements being to the more 3' J gamma genes as 11 alpha/beta+ clones had rearrangement(s) to JP1, the most 5' J gamma gene segment. Both alpha/beta+ and gamma/delta+ clones showed a similar pattern of V gamma gene usage in rearrangements to J gamma 1 or J gamma 2 with a lower proportion of the more 3' genes being rearranged to J gamma 2 than for the more 5' genes. Several alpha/beta+ and several gamma/delta+ clones had noncoordinate patterns of rearrangement involving both C gamma 1 and C gamma 2. Eleven out of fourteen CD8+ clones tested had both chromosomes rearranged to C gamma 2 whereas all clones derived from CD4-8- cells and having unconventional phenotypes (CD4-8- or CD4+8+) had at least one C gamma 1 rearrangement. Twelve out of twenty-seven CD4+ clones also had this pattern, suggesting that CD4-8+ clones had a tendency to utilize more 3' J gamma gene segments than CD4+ clones. There was some evidence for interdonor variation in the proportions of TCR gamma rearrangements to C gamma 1 or C gamma 2 in alpha/beta+ clones as well as gamma/delta+ clones. The results illustrate the unique nature of the V9JP rearrangement in gamma/delta+ clones and the possible use of a sequential mechanism of TCR gamma gene rearrangements during T cell differentiation is discussed. PMID- 2553275 TI - Phenotypic and functional differences in NK and LAK cells in the peripheral blood of sooty mangabeys and rhesus macaques. AB - Greater than 75% of the sooty mangabey monkeys at the Yerkes Regional Primate Research Center are naturally infected with SIV without any apparent clinical symptomology. On the other hand, experimental infection of rhesus macaques with SIV results in a clinical syndrome similar to human AIDS. These differences with regard to SIV infection prompted us to examine the natural immunosurveillance system of peripheral blood mononuclear cells (PBMC) from SIV-infected and uninfected monkeys of these two species. Phenotypic and functional studies of precursor and effector NK and LAK cells in the PBMC from these two species were carried out using monoclonal reagents, flow microfluorometry (FMF), and the standard in vitro 51Cr release assay against prototype K562 (NK sensitive) and RAJI (NK resistant, LAK susceptible) target cell lines. Data indicate that both NK and LAK cell activities in the PBMC of sooty mangabeys were significantly (P less than 0.01) greater than those in rhesus macaques. The predominant NK effector cells and LAK cell precursors were shown to be Leu 19-CD8+ in the PBMC of sooty mangabeys and Leu19+ CD8- in the PBMC of rhesus macaques as determined by panning depletion techniques and FMF analysis. On the other hand, the predominant LAK effector cells were found to be dual marked Leu 19+ CD8+ in rhesus macaques and Leu 19- CD8+ in sooty mangabeys. These qualitative and quantitative differences were not due to SIV infection of these two species since PBMC from both SIV-seropositive and virus-positive and SIV-sero-negative and virus-negative monkeys gave similar results. Moreover, of importance is the finding that the functional NK and LAK precursor cells are CD8+ and CD8- in sooty mangabeys and rhesus macaques, respectively. These data may have implications for the natural SIV/SMM virus-positive asymptomatic state of sooty mangabeys and may provide useful tools for tracing the ontogeny and lineage derivation of NK and LAK cells. PMID- 2553276 TI - Beta-adrenergic stimulation of phagocytosis in the unicellular eukaryote Paramecium aurelia. AB - Bete-adrenergic agonists isoproterenol and norepinephrine enhanced phagocytosis in Paramecium. Stimulation was stereospecific, dose-dependent and inhibited by the beta-agonists propranolol and alprenolol. Phorbol ester and forskolin potentiated the stimulatory effect of catecholamines on Paramecium phagocytosis. The dansyl analogue of propranolol (DAPN) was used for fluorescent visualization of the beta-adrenergic receptor sites in Paramecium which have been found to be localized at the cell membrane and within the membrane of the nascent digestive vacuoles. The appearance of the characteristic fluorescent pattern has been blocked by 1-propranolol. PMID- 2553277 TI - Functional rescue of temperature-sensitive defects in the cell-cycle mutants of rat 3Y1 fibroblasts by the small t-antigen deletion mutant of simian virus 40. AB - We examined the effects of large T antigen of simian virus 40 (SV40) on the proliferation phenotypes of temperature-sensitive (ts) mutants of rat 3Y1 fibroblasts, which cease proliferating in the G1 phase of the cell cycle at a restrictive temperature (39.8 degrees C). Four ts mutants, each representing independent complementation groups, were transformed with the dl-884 mutant of SV40 which lacks the unique coding region for small t antigen. In the case of two ts mutants, their transformed derivatives did not cease proliferation at 39.8 degrees C. In the other two mutants, the transformed cells continued to enter the S phase but the cells became detached from the dishes thereafter, at 39.8 degrees C. The proliferation phenotypes of the dl-884-transformed cells at 39.8 degrees C were quite similar with those of the same mutants transformed with the wild-type SV40. These results indicate that large T antigen alone is sufficient to overcome the inhibition of cellular entry into S phase caused by four different ts defects and determines the proliferation phenotypes of the cells after entering the S phase at a restrictive temperature, and that small t antigen does not alter the cellular phenotypes determined by large T antigen. PMID- 2553278 TI - Early events related with the behaviour of Trypanosoma cruzi within an endocytic vacuole in mouse peritoneal macrophages. AB - Transmission electron microscopy was used to analyse the process of interaction of Trypanosoma cruzi with resident and activated mouse peritoneal macrophages. Initially, the parasites are located within a membrane-bounded endocytic vacuole. Lysosomes from the host cell fuse and discharge their content into the parasite containing vacuole, as visualized by localization of horseradish peroxidase and acid phosphatase activity. Acridine orange was used to label secondary lysosomes in order to quantify the process of lysosome-phagosome fusion by fluorescence microscopy. The fusion index was higher for amastigote than for epimastigote and trypomastigote forms. Images were obtained showing that a few hours after ingestion of trypomastigote forms by the macrophages there is progressive disruption of the membrane lining the vacuole, until its complete disappearance. PMID- 2553280 TI - [A preliminary study on the relation of peripheral blood lymphocyte beta adrenoceptors and airway reactivity in asthmatics]. AB - The beta-adrenoceptors on intact peripheral blood lymphocytes were studied in 21 allergic asthmatics. 11 TDI asthmatics and 17 normal subjects with radioligand binding assay technique. The airway responsiveness to methacholine provocation test was determined. The results indicated that beta-adrenoceptor Bmax values in allergic and TDI asthmatics were reduced but the affinity between receptor and ligand was increased there was a positive correlation between the values of Bmax and FEV1%, V25/Pr in both asthmatic groups, a positive correlation between the values of Bmax and PC20 in allergic asthmatics. The beta-adrenoceptor responsiveness was remarkably reduced in allergic asthmatics. PMID- 2553279 TI - Effects of hydroxyurea on the phagocytosis of microspheres in V79 cells. AB - Phagocytosis of fluorescent microspheres in V79 cells arrested in the S phase by hydroxyurea (HU) was investigated by flow cytometry. The phagocytic activity increased with the exposure time of HU. No dependency on the HU concentration was observed in the concentration used. Cell size (projection area) and membrane fluidity, two factors effected by HU treatment, were examined in relation to phagocytic activity, and were found to be significantly increased. The elevation in phagocytic activity could be interpreted in terms of the alteration in the two physiological factors. PMID- 2553281 TI - [Occupational toluene diisocyanate (TDI) asthma--inhalation challenge test and TDI-HSA (human serum albumin) skin test and detection of specific IgE]. AB - Using a self-made bag inhalation challenge device. We diagnosed 12 TDI asthmatics, who were from 18 symptomatic workers. Of the 12 cases there were three types of airway reaction to TDI: three showed an immediate response; six, a late response; three, a dual response. After inhalation challenge, FEV1 PEFR, V50, and V25 descended obviously, suggesting that airway response to TDI in TDI asthmatics might occur in either larger or small airway. As a result of methacholine challenge nonspecific bronchial reactivity in 12 TDI asthmatics was markedly increased. After inhalation of TDI, the further descending of PC20 FEV1 showed that TDI could increase airway reaction to methacholine. Patients received skin test with TDI-HSA conjugate. There were positive response in 11 of the 12 TDI asthmatics and in 11 of the 62 TDI workers who had no symptoms after exposure to TDI, and no positive response in 18 common asthma patients. Therefore, TDI-HSA skin test can be used to a assist diagnosis of TDI asthma and a differential diagnosis from ordinary asthma. Specific IgE antibody levels showed no difference between TDI asthma and normal control group before TDI challenge. A marked increase in TDI asthma occurred after TDI challenge. PMID- 2553282 TI - Daily modifications of 3H-naloxone binding sites in the rat brain: a quantitative autoradiographic study. AB - The endogenous opioid peptides, the opiate receptors and several related behaviours, like opioid-mediated analgesia, show daily variations in different animal species including rats. The attempt to correlate the daily rhythm of opiate receptors in the central nervous system (CNS) to opiate related rhythmic phenomena requires an experimental approach with a high anatomical resolution, as the opioid distribution is very heterogeneous. In this paper we present the study of daily variations of 3H-naloxone binding sites in the different regions of the adult male rat brain, performed by means of quantitative autoradiography. Five rats are sacrificed at each investigated time of the day (0200, 0600, 1000, 1400, 1800 and 2200). The ligant is 3H-naloxone (4 nM), the quantification is performed by means of densitometric procedures (image analyzer Tesak VDC 501, computer Digital PDP 11, 3H-microscale). The statistical analysis is performed according to the single Cosinor method and the one-way analysis of variance followed by the multiple range test of Duncan. We analysed 33 different regions of the rat CNS, and the daily variations of opiate receptors are regionally selective. A circadian rhythm is found in the anterior cingulate cortex, hippocampal cortex, periventricular, medial, ventral, reticular and posterior nuclei of the thalamus, rhomboid, gelatinosus and rheuniens nuclei, lateral hypothalamus, locus coeruleus, grey substance of the pons, reticular formation of medulla oblongata, inferior olivary complex, medial part of the nucleus of the solitary tract and nucleus of the spinal tract of the trigeminal nerve. An ultradian rhythm is found in the medial and lateral preoptic areas, in the medial hypothalamus, in the medial and in the lateral nuclei of habenula. No significant variations during 24 hr according to the Cosinor analysis are found in the dorsal and lateral cerebral cortex, striatum, globus pallidus, bed nucleus of the stria terminalis, septal nuclei, lateral nucleus of the thalamus, cochlear nuclei, nucleus of the solitary tract, lateral and caudal parts, dorsal motor nucleus of the vagal nerve, XII and IX nerve nuclei. The amplitude of the daily variations observed ranges from 10 to 40%. Our results demonstrate the high anatomical selectivity of the daily modifications of 3H-naloxone binding sites in the rat CNS. They also indicate that quantitative autoradiography is a suitable and sensitive technique for these studies. PMID- 2553283 TI - Phosphatase-inhibitory activity and activation of murine macrophages by new 5' nucleotidase inhibitors, NPF-86IA, NPF-86IB, NPF-86IIA and NPF-86IIB. AB - New 5'-nucleotidase-inhibitory polyphenols named NPF-86IA, NPF-86IB, NPF-86IIA and NPF-86IIB were isolated from the seeds of Areca catechu L. The ability of the inhibitors to precipitate gelatin was investigated by microturbidimetry. These inhibitors produced weak turbidity. As 5'-nucleotidase is a kind of phosphatase, we examined the effects of these inhibitors on alkaline and acidic phosphatases. While they showed moderate inhibitory effects on the activity of acidic phosphatases, they did not have any significant effect on the activity of alkaline phosphatase. Therefore, they showed a higher inhibitory effect on the 5' nucleotidase than the other phosphatases, Murine macrophages were directly stimulated by the 5'-nucleotidase inhibitors. PMID- 2553284 TI - Enzymatic and chemical stability of 2',3'-dideoxy-2',3'-didehydropyrimidine nucleosides: potential anti-acquired immunodeficiency syndrome agents. AB - The enzymatic and chemical stability of three 2',3'-dideoxy-2',3' didehydropyrimidine nucleosides has been studied. Chemical degradtion of the analogues was measured in the pH range of 1.0-9.0. 2',3'-Dideoxy-2',3' didehydrocytidine (DDCN) degraded rapidly under acidic conditions, but the chemical stability was greater under basic conditions. The chemical degradation of 2',3'-dideoxy-2',3'-didehydrouridine (DDUN) and 2',3'-dideoxy-2',3' didehydrothymidine (DDTN) was not pH dependent and was faster than that of cytarabine. Enzymatic degradation of DDCN, DDUN and DDTN was not observed in human plasma, though cytarabine was degraded enzymatically under the same conditions. DDCN was also not degraded in the presence of mouse kidney cytidine deaminase. PMID- 2553285 TI - [Studies on mechanism of carcinogenesis in cell culture. III. Butyric acid induced phenotypic reversion of malignant transformed Syrian hamster lung cells (BHLB4)]. AB - The MNNG transformed Syrian hamster lung fibroblast cell line (BHLB4) established in our laboratory was treated with 2.0 mmol/L butyric acid. We obtained a butyric acid-dependent partially reversed cell line (ButB4) similar to normal cells in morphology, membrane properties, proliferation rate, colony-forming ability on solid agar culture, etc. However, when transplanted into nude mice, this cell line's tumorigenicity remained intact. Detection of cAMP levels in the cytoplasm showed much higher levels in ButB4 cells than in transformed cells. Butyric acid activity is evidently associated with changes of cAMP. In addition to phenotypical analysis, the karyotypes of ButB4 and BHLB4 were analyzed in detail. It was found that the disomy rate of C15 was significantly increased in ButB4 cells (73%) as compared with BHLB4 cells (33%). These results show that butyric acid not only induces phenotypic reversion in BHLB4 cells directly, but can also selectively kill C15 monosomy cell clones, allowing rapid growth of C15 disomy cell clones. Butyric acid may also influence maintenance of the reversion phenotype. PMID- 2553286 TI - [Studies on the physical state of human papillomavirus 16 DNA in biopsies of patients with cervical carcinoma]. AB - Five biopsies from cervical cancer containing human papillomavirus (HPV) 16 DNA sequences confirmed by dot blot analysis were used to study the physical state of HPV 16 DNA in cervical cancer. Southern blots prepared after: 1) sample DNA was cleaved with restriction enzymes which do not digest HPV 16;2) sample DNA was cleaved with restriction enzymes (pstI) capable of digesting HPV 16 DNA; 3) sample DNA was not digested with restriction enzymes; and 4) two-dimensional agarose gel electrophoresis was performed on sample DNA digested with enzymes which do not cleave HPV 16 DNA. It was shown that HPV 16 DNA was integrated into the cellular genome. PMID- 2553287 TI - [Cryptosporidium and candida in pediatric diarrhea in Abidjan]. AB - In a study involving 104 children hospitalized with diarrhoea, 9% were infected with oocyst Cryptosporidium spp. add 56% with such yeast-fungus as Candida (C. Candida 38%). The manifestations noted in cryptosporidiosis infected children are acute diarrhea, vomiting and hyperthermia. One subject out of five who were tested for antibody to HIV appeared to be antibody positive. The patients immunity from the disease was not checked. A mycological test must be systematically carried out in case of children diarrheal outbreak. PMID- 2553288 TI - [Epidemiology of toxoplasmosis in Ivory Coast]. AB - The purpose of this study was to determine the prevalence of toxoplasmosis in samples of healthy populations in Ivory Coast. Various factors have been studied: age, sex, ethnic group, occupation, food, contacts with animals, climate. The samples were obtained in four places which were different by geographical situation and climate: Abidjan (urban site), Bonoua (littoral site), Bin-Houye (forest site), Odienne (predesert site). The samples of sera or blood were deposited on filter-papers and tested by latex agglutination (for IgG) and ISAGA (for IgM). Results were analysed by statistical methods. More than 2,000 samples have been studied, showing a prevalence higher in humid tropical zones (55.6% to 70%) than in predesert zone (37.2%). The first contact of the child with the parasite was more often before reaching 10 years but the risk for women was important until the age of 20. The other studied factors didn't appear to have any effect. PMID- 2553289 TI - Inhibition by lipoxygenase inhibitors of 7-bromomethylbenz[a]anthracene-caused epidermal ornithine decarboxylase induction and skin tumor promotion in mice. AB - 7-Bromomethylbenz[a]anthracene (BrMBA) has been shown to have a tumor-promoting action in mouse skin without an initial direct interaction with protein kinase C, which is believed to be a receptor for phorbol ester tumor promoters such as 12-O tetradecanoylphorbol-13-acetate (TPA). An application of BrMBA to mouse dorsal skin caused epidermal ornithine decarboxylase (ODC) induction in a dose-dependent manner with a peak of activity at 12 h after the application. A single topical application of BrMBA failed to induce mouse ear edema formation, i.e. inflammation. However, repeated applications of BrMBA, i.e. twice a week for 3-4 times, caused a significant edema. Unlike TPA, BrMBA failed to stimulate the superoxide anion generation of rabbit peritoneal polmorphonuclear leukocytes. Lipoxygenase inhibitors such as 3,4,2',4'-tetrahydroxychalcone, nordihydroguaiaretic acid, quercetin and 2,3,5-trimethyl-6-(12-hydroxy-5,10 dodecadiynyl)-1,4-benzoquinone (AA861) effectively inhibited BrMBA-caused epidermal ODC induction and ear edema formation. In addition, BrMBA-caused skin tumor promotion was also potently inhibited by 3,4,2'4'-tetrahydroxychalcone and quercetin. These results indicate that a mechanism susceptible to lipoxygenase inhibitors plays a role not only in the TPA-caused but also in the BrMBA-caused epidermal ODC induction, skin inflammation and tumor promotion. It seems unlikely that superoxide anion generation is involved in the mechanism of BrMBA-caused skin tumor promotion. PMID- 2553290 TI - Epoxidation of 7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene via a hydroperoxide dependent mechanism catalyzed by lipoxygenases. AB - The lipoxygenase catalyzed epoxidation of 7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene (BP-7,8-diol) was examined. Epoxidation of the BP-7,8-diol was catalyzed by 5- and 15-lipoxygenase in the presence of either arachidonic acid, gamma-linolenic acid, or 15-hydroperoxyeicosatetraenoic acid (15-HPETE). The anti-9,10-epoxy-7,8 dihydroxy-7,8-dihydrobenzo[a]pyrene isomer was formed in greater quantities than the syn isomer, indicative of peroxyl radical mediated epoxidation. Epoxidation was dependent on time, enzyme and fatty acid concentration. There was no difference in the time course of epoxidation with either arachidonic acid or 15 HPETE, although the initial rate of oxygen consumption was approximately 55-fold greater with arachidonic acid. The lipoxygenase inhibitor and anti-oxidant nordihydroguaiaretic acid inhibited epoxidation in a dose-dependent manner in incubations initiated with either arachidonic acid or 15-HPETE. The anti-oxidant butylated hydroxyanisole also inhibited the epoxidation. Incubations conducted under anaerobic conditions with 15-lipoxygenase and either arachidonic acid or 15 HPETE significantly decreased epoxidation. This suggests that the oxygen inserted into BP-7,8-diol is derived from the atmosphere. The epoxidizing peroxyl radicals could not be detected but their precursors, carbon-centered radicals, were detected by using the ESR spin trapping technique in incubations of 15 lipoxygenase with 15-HPETE. This radical, formed by reduction and rearrangement of the hydroperoxide, may trap oxygen to form a peroxyl radical. We propose that the epoxidizing species is a peroxyl radical derived from 15-HPETE rather than from arachidonic acid. This proposal is based on the similar amounts of epoxidation, but dissimilar amount of oxygen consumed with both fatty acids. Since lipoxygenases are widely distributed in vivo, especially in areas where tumors arise such as the pulmonary epithelium, peroxyl radical formation by these enzymes may have an important role in chemical carcinogenesis. PMID- 2553291 TI - Characterization of A1 adenosine receptors in atrial and ventricular myocardium from diseased human hearts. AB - The purpose of the present study was to characterize adenosine receptors in human atrial and ventricular myocardium. In isolated electrically driven preparations, adenosine produced "direct" negative inotropic effects in atrial myocardium (AT). In ventricular myocardium (VE), it only had negative inotropic properties when force of contraction had been stimulated with isoprenaline ("indirect" effect), but it has no inotropic effect alone. The adenosine receptor antagonist 8 phenyltheophylline antagonized the "direct" and "indirect" effects; these findings indicated that both effects were mediated by adenosine receptors. In cardiac membranes from human AT and VE, adenosine receptors were characterized with [3H]-8-cyclopentyl-1,3-dipropylxanthine (DPCPX) binding. The effects of agonists R-(-)-N6-phenylisopropyladenosine (R-PIA), S-(+)-N6 phenylisopropyladenosine (S-PIA), and 5'-(N-ethylcarboxamido) adenosine (NECA) and the effects of guanine nucleotides [Gpp(NH)p] were studied also. The antagonist affinities as judged from the apparent affinity, Kd, of [3H]DPCPX were similar in AT (2.2 nmol/l; 95% confidence limits, 1.4-3.7) and VE (1.8 nmol/l; 95% confidence limits, 1.0-3.0). The number of adenosine receptors was 1.7 times greater in AT (26.9 +/- 2.33 fmol/mg protein; n = 5) than in VE (16.2 +/- 2.3 fmol/mg protein; n = 5). High and low affinity states of adenosine receptors evaluated with the influence of Gpp(NH)p on agonist competition with R-PIA were similar in AT or VE. The rank orders of potency for agonists (R-PIA greater than S-PIA greater than NECA) and antagonists (DPCPX greater than 8-phenyltheophylline greater than theophylline) were characteristic for the A1 receptor subtype. It is concluded that A1 adenosine receptors exist in the human myocardium. Since binding properties were similar in AT and VE, the same A1 adenosine receptor probably couples to different effectors in a similar guanine nucleotide-dependent way. [3H]DPCPX is the first radiolabeled antagonist ligand that allows detection of A1 adenosine receptors and their coupling in the human myocardium. PMID- 2553292 TI - Blockade of cardiac sodium channels by lidocaine. Single-channel analysis. AB - The mechanism of interaction of lidocaine with cardiac sodium channels during use dependent block is not well defined. We examined the blockade of single cardiac sodium channels by lidocaine and its hydrophobic derivative RAD-242 in rabbit ventricular myocytes. Experiments were performed in cell-attached and inside-out patches. Use-dependent block was assessed with trains of ten 200-msec pulses with interpulse intervals of 500 msec and test potentials of -60 to -40 mV. Single channel kinetics sometimes showed time-dependent change in the absence of drug. During exposure to 80 microM lidocaine, use-dependent block during the trains was associated with a decrease in the average number of openings per step. At -60 mV, mean open time was not significantly changed (control, 1.4 +/- 0.6 msec; lidocaine, 1.2 +/- 0.3 msec, p greater than 0.05). Greater block developed during trains of 200-msec pulses compared with trains of 20-msec pulses at the same interpulse interval at test potentials during which openings were uncommon later than 20 msec (-50 and -40 mV). Prolonged bursts of channels showing slow-gating kinetics were observed both in control and the presence of 80 microM lidocaine. However, lidocaine may decrease the late sodium current by altering the kinetics of slow gating. The hydrophobic lidocaine derivative RAD-242, which has a 10-fold greater lipid solubility than lidocaine, decreased the peak averaged current during pulse train stimulation by 60% without a change in the mean open time. Our results suggest that the major effect of lidocaine during use-dependent block involves the interaction with a nonconducting state of the sodium channel followed by a failure to open during subsequent depolarization. PMID- 2553293 TI - Protection by superoxide dismutase from myocardial dysfunction and attenuation of vasodilator reserve after coronary occlusion and reperfusion in dog. AB - Previous studies indicate impairment of coronary arterial ring relaxation and loss of coronary vasodilator reserve after coronary artery occlusion and reperfusion. These changes are mediated in part through loss of endothelium derived relaxing factor (EDRF) and/or myocardial neutrophil accumulation. To examine if superoxide dismutase (SOD), a scavenger of superoxide radicals, would modify the diminished coronary vasodilator reserve after temporary coronary occlusion in the intact animal, open-chest mongrel dogs were subjected to 1 hour of circumflex (Cx) coronary artery occlusion followed by 1 hour of reperfusion and treated with saline or SOD. Before Cx occlusion, coronary blood flow increased, and vascular resistance decreased (both p less than 0.01) in response to EDRF-dependent vasodilator acetylcholine as well as EDRF-independent vasodilator nitroglycerin. After Cx reperfusion, resting Cx coronary blood flow and vascular resistance were similar to the preocclusion values. In the saline treated animals, there was evidence of myocardial dysfunction, which was measured by segmental shortening (-6 +/- 2% vs. 10 +/- 2%). Furthermore, increase in Cx coronary blood flow and reduction in vascular resistance in response to both vasodilators were significantly (p less than 0.01) impaired; these occurrences suggested loss of coronary vasodilator reserve. Myocardial histology showed extensive capillary plugging by neutrophils in the Cx-supplied myocardium. Myocardial myeloperoxidase activity, an index of neutrophil infiltration, was also increased in the Cx compared with the left anterior descending coronary artery region (p less than 0.02). Treatment of dogs with SOD, started at the end of Cx occlusion and continued during reperfusion, exerted significant (p less than 0.01) protective effect against reperfusion-induced attenuation of coronary vasodilator reserve in response to both acetylcholine and nitroglycerin. Loss of myocardial function (segmental shortening 5 +/- 1% vs. 10 +/- 1%) was less than in the saline-treated animals (p less than 0.01). Cx region-myocardial neutrophil accumulation and myeloperoxidase activity were also less (p less than 0.02) in the SOD-treated than in the saline-treated dogs. These observations suggest that coronary reperfusion impairs coronary vasodilator reserve in intact dogs. This impairment can be modified by treatment of animals with SOD before reperfusion. Capillary plugging by neutrophils may contribute to the altered coronary vasodilator reserve observed in the immediate postreperfusion period, and SOD modifies this reperfusion-induced impairment. PMID- 2553294 TI - Stereoselective block of cardiac sodium channels by RAC109 in single guinea pig ventricular myocytes. AB - The effects of the optical stereoisomers of the local anesthetic RAC109 (RAC109-I and RAC109-II) on sodium current in isolated guinea pig ventricular myocytes were investigated by use of the whole-cell variation of the patch-clamp technique. RAC109-I and RAC109-II produced similar levels of tonic block, but RAC109-I produced a significantly larger use-dependent block on repetitive pulsing to potentials positive to -60 mV. Definition of the time courses of block development at -20 mV and recovery at -140 and -160 mV indicated that RAC109-I had a higher affinity for activated and inactivated channels and dissociated more slowly at hyperpolarized potentials compared with RAC109-II. Removal of fast inactivation by alpha-chymotrypsin intensified tonic block but did not reduce use dependent block by RAC109-I; this finding suggests that channel inactivation is not necessary for use-dependent block. The guarded-receptor model was used to calculate apparent rate constants of drug binding and unbinding. According to the model, RAC109-I and RAC109-II have significantly different unbinding rate constants for channels when they exist predominantly in rested, activated, or inactivated states, as well as significantly different binding rate constants when channels are activated. However, the apparent rates of drug binding to closed (rested and inactivated) channels are not significantly different for the two isomers; this finding indicates that drug binding to closed channels is not markedly stereospecific, in contrast to unbinding. The effects of RAC109 stereoisomers on cardiac sodium channels were also qualitatively similar to those previously reported in nerve; these findings suggest that the binding sites for local anesthetics in both tissue types have a similar structural topography. PMID- 2553295 TI - Comments on "Nonlinear Relation between Vmax and INa in Canine Cardiac Purkinje cells. PMID- 2553296 TI - Deleterious effects of oxygen radicals in ischemia/reperfusion. Resolved and unresolved issues. AB - Oxygen free radicals are known to be generated during periods of ischemia followed by reperfusion. There is still some controversy, however, concerning the use of electron paramagnetic resonance spectroscopy to accurately detect and identify the free radical species that are formed. There is no doubt that oxygen radicals are deleterious to the myocardium; free radicals cause left ventricular dysfunction and structural damage to myocytes and endothelial cells in both in vitro and in vivo preparations. Potential sources of these cytotoxic oxygen species include the xanthine oxidase pathway, activated neutrophils, mitochondria, and arachidonate metabolism, yet the crucial source of free radicals in the setting of ischemia and reperfusion is unresolved. There is little doubt that oxygen radicals play a role in the phenomenon of stunned myocardium induced by brief periods of ischemia followed by reperfusion; numerous studies have consistently observed that pretreatment with free radical scavengers and antioxidants enhances contractile function of stunned, postischemic tissue. Whether oxygen free radical scavengers administered only during reperfusion enhance recovery of stunned myocardium in models of brief ischemia remains to be determined. In models of prolonged ischemia (2 hours) followed by reperfusion, we have not observed a beneficial effect of scavengers on stunned myocardium. The issue of whether oxygen free radical scavengers are capable of reducing so-called irreversible or lethal reperfusion injury remains, in our opinion, unresolved. Although some studies have observed that agents such as superoxide dismutase and catalase reduce infarct size in ischemia and reperfusion models, many others have reported negative results. Additional studies will be needed to resolve this ongoing controversy. Oxygen free radicals may also contribute to reperfusion induced arrhythmias in rodent heart preparations; however, less data are available in other animal models. The concept of reperfusion injury should not be considered a deterrent to reperfusion for the treatment of acute myocardial infarcts in the clinical setting. Thrombolytic therapy reduces myocardial infarct size, enhances recovery of left ventricular function, and improves survival. Whether incremental beneficial effects on these parameters will be obtained when oxygen radical-scavenging agents are used as adjuvant therapy to thrombolysis in patients remains to be determined. PMID- 2553297 TI - End-stage dilated cardiomyopathy. Persistence of enterovirus RNA in myocardium at cardiac transplantation and lack of immune response. AB - Tissue from the explanted hearts of 21 patients with idiopathic dilated cardiomyopathy or 19 patients with other specific heart muscle diseases were investigated for presence of enterovirus-specific RNA with an enterovirus group specific cDNA probe. This was complementary to coxsackievirus B2 RNA sequences between nucleotide numbers 6,550 and 7,400, which are highly conserved between enteroviruses. Hearts from six patients with dilated cardiomyopathy and one patient with ischemic heart disease were found to contain virus RNA. Serology revealed that only one patient (dilated cardiomyopathy group) was positive for coxsackievirus B-specific IgM but negative for virus RNA in the myocardium. Quantitation of leukocytes and T-lymphocytes in the myocardium and expression of major histocompatibility locus antigens revealed no significant differences associated with persistence of virus RNA. These data demonstrate that enterovirus RNA persists in myocardium of a significant proportion of patients with end-stage dilated cardiomyopathy in the absence of a continuing cell-mediated or humoral immune response. PMID- 2553298 TI - Increased adenosine concentration in blood from ischemic myocardium by AICA riboside. Effects on flow, granulocytes, and injury. AB - Morbidity and mortality from acute coronary artery occlusion may be reduced if local myocardial adenosine concentration is augmented because 1) coronary collateral blood flow during ischemia increases with adenosine infusion, and 2) granulocytes that accumulate in the microcirculation during ischemia are, to a large extent, inhibited by adenosine from generating superoxide anion free radicals, from adhering to vascular endothelium, and from damaging endothelial cells in culture. Using a cultured lymphoblast model system, we found that 5 amino-4-imidazole carboxamide (AICA) riboside enhanced adenosine accumulation during ATP catabolism. Therefore, AICA riboside pretreatment was used in canine myocardium to selectively increase adenosine concentration in the ischemic area during 1 hour of ischemia. At 5 minutes of ischemia, endocardial flow to ischemic myocardium in saline-treated and AICA riboside-treated dogs was 0.06 +/- 0.03 and 0.34 +/- 0.11 ml/min/g, respectively (p less than 0.01); flow to nonischemic myocardium was not affected. Ventricular tachycardia and premature ventricular depolarizations were significantly attenuated in the AICA riboside-treated dogs. Blood pressure and heart rate were not affected by AICA riboside. In venous blood from ischemic tissue, adenosine increased from undetectable levels (less than 0.01 microM) to 0.22 +/- 0.08 microM in saline and 1.79 +/- 0.06 microM in AICA riboside-treated dogs, respectively (p less than 0.001). Coronary vein inosine concentrations were greater in saline than in AICA riboside-treated dogs. In separate in vitro studies, AICA riboside did not alter the removal rate of adenosine from canine blood. Indium-labeled granulocyte accumulation was significantly less in ischemic myocardium in AICA riboside-treated compared with saline-treated dogs. In addition, adenosine, but not AICA riboside, inhibited in vitro canine granulocyte superoxide production. We conclude that AICA riboside given before myocardial ischemia augments adenosine concentration, decreases arrhythmias, decreases granulocyte accumulation, and improves collateral flow to ischemic myocardium. One of the beneficial mechanisms could be an increased production of adenosine rather than inosine from ATP catabolism that causes vasodilation and inhibition of granulocytes. We propose a new hypothesis regarding regulation of the inflammatory reaction to ischemia in the microcirculation. Adenosine, in addition to its vasodilator action, is an anti injury autacoid that links ATP catabolism to inhibition of granulocyte adherence, microvascular obstruction, and superoxide anion formation. PMID- 2553299 TI - Action potential duration alternans in dog Purkinje and ventricular muscle fibers. Further evidence in support of two different mechanisms. AB - An abrupt shortening of cycle length causes action potential duration (APD) alternation in both canine Purkinje (P) and ventricular (V) muscle fibers. Our recent study suggested that APD alternans is determined by the process controlling APD during electrical restitution in P but not in V fibers. In the latter, alternans was attributed to changes in the availability of intracellular calcium [Ca2+]i. We examined this hypothesis further with the following pharmacologic probes known to alter restitution or action of [Ca2+]i: tetradotoxin (0.5-3.0 microM), lidocaine HCl (2.0-12.0 micrograms/ml), sotalol (10 microM), nicorandil (10-20 microM), 4-amino-pyridine (0.5 microM), ryanodine (10 microM), caffeine (2 mM), and ARL 115 BS (100 microM). Alternans in P fibers persisted under all studied conditions but varied in magnitude depending on the time constant and amplitude of restitution. In V fibers, the magnitude of alternans did not correlate with APD changes during restitution, and APD alternans was associated with the alternans of action potential shape and alternans of developed tension. Alternans in V was suppressed by caffeine at 2.0 mM [Ca2+]o when tension was increased and by ryanodine at 1.0 mM [Ca2+]o when tension was decreased. Alternans in V was not altered by changes in [Ca2+]o within the range of 1.0-4.0 mM; by ARL 115 BS, a compound that increases myofibrillar sensitivity to calcium; or by any other pharmacologic probes. We concluded that in P fibers, APD alternans was determined by the factors controlling APD in the absence of alternans; V fibers posses an independent mechanism of alternans linked to alternans of tension and controlled by [Ca2+]i; in V fibers, alternans could be suppressed by both positive and negative inotropic interventions; and calcium released from sarcoplasmic reticulum plays an important role in the V alternans. PMID- 2553300 TI - Singlet oxygen-induced arrhythmias. Dose- and light-response studies for photoactivation of rose bengal in the rat heart. AB - In a study of aerobically perfused rat hearts, the in situ photoactivation (530 590 nm) of rose bengal (a process that leads to the production of singlet oxygen and superoxide) has been shown to lead to the rapid development of electrocardiographic abnormalities and arrhythmias. With rose bengal concentrations of 1,000, 500, 250, 100, and 50 nmol/l (n = 6/group), photoactivation (3,600 lx) led to electrocardiographic changes (inversion of the T wave, Q-T prolongation, or both) after 3.8 +/- 0.9, 4.5 +/- 0.7, 11.8 +/- 2.1, 24.8 +/- 3.9, and 65.3 +/- 6.0 seconds), respectively; ventricular premature beats occurred in 100% of hearts after 0.5 +/- 0.2, 1.1 +/- 0.3, 2.2 +/- 0.7, 4.4 +/- 0.8, and 6.6 +/- 1.2 minutes, respectively. Ventricular tachycardia occurred in 83%, 83%, 83%, 67%, and 50% of hearts after 2.1 +/- 0.2, 2.1 +/- 0.4, 2.8 +/- 0.7, 5.7 +/- 2.0, and 11.2 +/- 1.9 minutes, respectively, and complete atrioventricular block in 100%, 100%, 100%, 100%, and 67% of hearts after 3.8 +/- 0.7, 6.5 +/- 1.0, 5.5 +/- 0.9, 13.8 +/- 1.0, and 14.1 +/- 0.9 minutes, respectively. With a fixed concentration (250 nmol/l) of rose bengal, similar light-response relations were observed. Photoactivation of rose bengal had no effect on heart rate but caused a transient (0-4 minutes) vasodilation followed by a progressive vasoconstriction. In further studies in which rose bengal was washed out for 10 minutes before photoactivation, several arrhythmias still developed, indicating that rose bengal binds strongly to tissue and acts as a cellular level rather than in the vascular compartment. To assess the reversibility of rose bengal-induced effects, hearts (n = 6/group) were perfused with rose bengal (250 nmol/l) for 1, 2, 4, 6, and 20 minutes followed by perfusion in the dark for 19, 18, 16, 14, and 0 minutes, respectively. During dark perfusion, the incidence of arrhythmias declined and any decrease in coronary flow was reversed. However, analysis of contents of adenosine triphosphate, creatine phosphate, lactate, and creatine kinase leakage indicated the occurrence of severe injury that did not abate on termination of photoactivation. Finally, although many arrhythmias developed before the onset of vasoconstriction, the reduction in flow with consequent ischemia was shown to exacerbate vulnerability to arrhythmias. In conclusion, short-lived reactive oxygen intermediates such as singlet oxygen and superoxide, which are produced during the photoactivation of rose bengal, can cause rapid and major damage to the heart and its function. PMID- 2553301 TI - Adequate preservation of right ventricular function after coronary sinus cardioplegia. A clinical study. AB - Whether retrograde coronary sinus cardioplegia adequately preserves right ventricular (RV) function is still a point of concern. Using technetium Tc 99m labeled red blood cells, we assessed global and segmental RV function by first pass and gated blood-pool radionuclide angiocardiography before and within 24 hours after aortic valve replacement in 14 consecutive patients (age, 58 +/- 5 years; mean +/- SEM). Coronary sinus cardioplegia was given in a multidose fashion at a flow rate of 50-70 ml/min through a balloon-tipped catheter, with the inflated balloon kept seated around the intra-atrial rim of the coronary sinus orifice. Additional myocardial protection was provided by systemic (25 degrees C) and topical hypothermia. Postoperatively, none of the patients had clinical or hemodynamic patterns suggestive of RV dysfunction. The postoperative global RV ejection fraction (0.49 +/- 0.03) was similar to the preoperative value (0.49 +/- 0.01). Analysis of segmental wall motion did not reveal postoperative abnormalities of new onset in any of the three anatomically defined RV regions (free wall, apex, and septum). Similarly, RV end-diastolic and end-systolic volume indexes (ml/m2) were not significantly affected by coronary sinus cardioplegia, being 71.6 +/- 5.8 and 36.1 +/- 3.5 before, and 67.4 +/- 3.8 and 34.5 +/- 2.3 after aortic valve replacement, respectively. We conclude that retrograde coronary sinus cardioplegia does not cause a detectable impairment of RV function if the balloon catheter does not obstruct the terminal tributaries of the coronary sinus and, hence, does not impede delivery of cardioplegia to right sided cardiac structures. PMID- 2553302 TI - Serum lipids and cholesterol metabolism during guar gum, plantago ovata and high fibre treatments. AB - Short-term viscous dietary fibres, plantago ovata and guar gum preparations, decreased serum cholesterol, mainly LDL cholesterol, as compared to low fibre or nonviscous high fibre periods, through enhancing cholesterol elimination as fecal bile acids. These changes were associated with significant increases in serum levels of cholesterol precursors, both in methylsterols and demethylated precursor sterols, while that of cholestanol (saturated cholesterol derivative) was decreased. In addition, cholesterol precursor contents were positively related, cholestanol negatively, to fecal cholesterol elimination both on and off viscous fibres. These findings indicate increased cholesterol synthesis, also seen by sterol balance data. As compared to low fibre diet, nonviscous high fibre diet conserved bile acids and decreased cholesterol absorption. Thus, it had no effect on cholesterol synthesis as indicated by fecal total steroids or serum levels of cholesterol precursors. PMID- 2553303 TI - Increased furosemide-sensitive sodium influx into erythrocytes in non-insulin dependent diabetes mellitus associated with hypertension. AB - Our previous study demonstrated increased sodium (Na+) influx into ouabain treated erythrocytes in diabetic patients with hypertension. The present study was designed to estimate the effect of furosemide on the Na+ influx in non insulin-dependent diabetes mellitus (NIDDM) with hypertension. Both Na+ influx in the absence of furosemide (total Na+ influx; TNaI) and furosemide-sensitive Na+ influx (FSNaI) were increased in patients with essential hypertension (group 1, n = 22) than those in non-diabetic, normotensive control subjects without family history of hypertension (control group, n = 26, P less than 0.001 and P less than 0.001, respectively). TNaI and FSNaI in diabetic patients without hypertension (group 2, n = 28) were also higher than those in the control group (P less than 0.02 and P less than 0.001, respectively), but no significant differences were found in TNaI or FSNaI between groups 1 and 2. TNaI and FSNaI in diabetic patients with hypertension (group 3, n = 19) were elevated than those in control group (P less than 0.001 and P less than 0.001, respectively), in group 1 (P less than 0.05 and P less than 0.02, respectively) and in group 2 (P less than 0.001 and P less than 0.001, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553304 TI - Partial purification and characterization of human hepatoma-derived growth factor. AB - A human hepatoma cell line, HuH-7, which was established from a hepatoma tissue removed at surgical operation from an adult Japanese male, grows autonomously in a serum-free chemically defined medium. A human hepatoma-derived growth factor with potent growth-promoting activity, was partially purified from the conditioned medium of HuH-7 cells, using ion-exchange, heparin affinity chromatography and gel filtration. The partially purified growth factor (HuHGF) is acid- and heat-labile, and sensitive to reduction and trypsin digestion. HuHGF, which is of relatively high molecular weight (about 64 kDa) examined by gel filtration, stimulates not only the DNA synthesis of Swiss mouse 3T3 fibroblasts, but also the growth of HuH-7 cells in serum-free chemically defined medium, suggesting that it is produced by and acts on HuH-7 cells as an autocrine growth factor. This putative growth factor may play an important role in the autonomous growth of hepatoma and may lead to useful diagnostic or therapeutic approaches to human hepatoma. PMID- 2553305 TI - Increased acylphosphatase levels in erythrocytes from hyperthyroid patients. AB - Acylphosphatase activity and content were measured in erythrocytes from hyperthyroid patients and healthy controls. In addition, the soluble enzymes glucose-6-phosphate dehydrogenase, hexokinase, and the membrane bound (Na+ + K+) ATPase and Ca2+-ATPase were assayed. Our results confirmed previous studies indicating a decrease of (Na+ + K+)-ATPase and an increase of Ca2+-ATPase activity in hyperthyroid erythrocytes. While glucose-6-phosphate dehydrogenase was not significantly changed, hexokinase and acylphosphatase activities were significantly higher in the hyperthyroid group. Both activities and content of acylphosphatase returned to normal levels in erythrocytes from treated patients, when they were euthyroid. These findings suggest that an excess of thyroid hormones may stimulate acylphosphatase biosynthesis in erythroid cells and indicate a potential clinical usefulness of this enzyme in hyperthyroidism. PMID- 2553306 TI - Body fluid variations and endogenous digitalis-like compounds during chronic NaCl loading in Wistar rats. AB - Circulating digitalis-like compounds have been proposed to be raised in volume expanded hypertension and to participate in Na+ homeostasis. We have investigated the temporal relationships between the activity of these circulating digitalis like compounds, blood pressure and body fluid volume variations during a chronic NaCl load in the Wistar rat. Characteristics of salt-loaded rats were compared to those of weight-matched controls. At one week, when extracellular fluid volume (ECFV) was elevated, the capacity of plasma extracts to inhibit the Na+K+ATPase activity begun to rise. At two weeks, ECFV remained elevated, and plasma volume, blood pressure and the activity of plasma digitalis-like compounds increased. After 13 weeks, the continuous rise in plasma digitalis-like activity and in blood pressure was accompanied by the return of body fluid volumes towards control values. These changes in plasma digitalis-like activity, body fluid volumes, and systolic blood pressure during a high NaCl diet are compatible with the proposed role of circulating digitalis-like compounds as natriuretic and hypertensive factors. PMID- 2553307 TI - Evidence for non-lysosomal storage of N-acetylneuraminic acid (sialic acid) in sialuria fibroblasts. AB - The results of the investigations reported here indicate that patients affected with the infantile sialic acid storage disorder (ISSD) and the original French sialuria patient suffer from distinct and fundamentally different disorders. While phase microscopy and immunochemical studies demonstrated abnormal storage within intracellular inclusions in ISSD cells, no morphological evidence of storage within any subcellular organelles was found in the sialuria cells. Moreover, comparative subcellular fractionation studies on gradients of colloidal silica showed the excess sialic acid in ISSD cells to be located within the light (buoyant) lysosomal fraction, while the excessive, free sialic acid in the sialuria cells was found in the cytoplasmic fraction with no increased storage within the lysosomal fractions. It is concluded that the sialic acid abnormalities in ISSD and the French type of sialuria are the result of very different biochemical and genetically unrelated abnormalities. PMID- 2553308 TI - Variation in lymphoproliferative responses during recrudescent orofacial herpes simplex virus infections. AB - Sixty-five patients with recrudescent orofacial herpes simplex virus (HSV) infections all had circulating HSV-specific antibody measured by ELISA and cell mediated immunity (CMI) to HSV by in vitro lymphoproliferation. Thirteen control subjects with no history of HSV were negative for both tests. Thirty-three patients, repeatedly investigated during 6 to 38 months, had between 1 and 8 recrudescences each. Lymphoproliferative responses to HSV were low during recrudescence, rose to a peak a few weeks later and then declined to a positive background level. However, ELISA titres and lymphoproliferative responses to concanavalin A were high throughout, and peripheral blood mononuclear cell (PBMC) subset numbers measured by fluorescent flow cytometry remained within normal limits. During HSV lesions, depressed lymphoproliferation to HSV was abrogated by removal of CD8+ T cells from PBMC either by using a panning technique (nine patients) or by cell sorting (three patients). Reconstitution of the CD8-depleted population suppressed the lymphoproliferative response to HSV. Depletion of CD8+ T cells did not affect lymphoproliferation to HSV outwith recrudescence (four patients), nor lymphoproliferative responses to another antigen (PPD; five patients) during recrudescence. Thus, reduced lymphoproliferation to HSV during recrudescence may be due to HSV-specific CD8+ suppressor T lymphocyte function, rather than lack of HSV-responsive lymphocytes. This may result in depression of normal CMI responses to the virus during an asymptomatic recurrence allowing recrudescent lesions to develop. PMID- 2553310 TI - Centrally mediated increased sympathetic activity is not important in the genesis of ACTH-induced hypertension in sheep. AB - 1. Adrenocorticotrophin (ACTH) administration to sheep produces a rapid adrenally dependent hypertension which is maximal after 3 days and associated with increased cardiac output (CO) and heart rate (HR), while calculated total peripheral resistance remains unchanged. 2. This study investigated the proposal that a centrally mediated increase in sympathetic activity is important in the development of ACTH-induced hypertension. 3. Concomitant intravenous infusions of either clonidine (60 micrograms/kg per day) or methyldopa (60 mg/kg per day) with ACTH (5 micrograms/kg per day) failed to inhibit the increase in mean arterial pressure (MAP) observed with ACTH. 4. In a separate experiment clonidine abolished the increase in CO and HR but not the pressor response associated with ACTH administration. 5. These results do not support a role for centrally mediated increase in sympathetic activity in the genesis of ACTH-induced hypertension. PMID- 2553309 TI - Antigen-stimulated human interferon-gamma generation: role of accessory cells and their expressed or secreted products. AB - In response to cytomegalovirus (CMV) and Toxoplasma gondii antigens, T4+ cells from seropositive donors produce interferon-gamma (IFN-gamma) by different mechanisms; one (T. gondii) dependent upon and the other (CMV) largely independent of interleukin-2 (IL-2) and its receptor. To determine whether IFN gamma-generating mechanisms unrelated to IL-2 also differ, we examined the requirement for accessory cells and their expressed or secreted products. In response to both specific antigens, IFN-gamma secretion was strictly dependent upon the presence of accessory cells (monocytes), and was largely inhibited by monoclonal antibodies to class II (HLA-DR and -DQ) but not class I MHC antigens. Both CMV and T. gondii antigens stimulated monocytes to release interleukin-1 (IL 1), and IFN-gamma production in response to both antigens was abolished by pretreatment with anti-IL-1 antibody. In contrast, the secretion of tumour necrosis factor (TNF) was not stimulated by either antigen, and IFN-gamma production was not diminished by antisera directed at TNF-alpha or TNF-beta. We conclude that CMV and T. gondii antigen-induced IFN-gamma production requires a similar accessory cell mechanism, and that soluble antigen-stimulated IFN-gamma secretion by human T4+ cells is dependent on monocytes, expression of class II MHC antigens, and the presence of IL-1. PMID- 2553311 TI - Production of interleukin 1-like factor from human peripheral blood monocytes and polymorphonuclear leukocytes by superoxide anion: the role of interleukin 1 and reactive oxygen species in inflamed sites. AB - In the present study, we investigated the possibility that oxidative stress to human peripheral blood monocytes and polymorphonuclear leukocytes (PMNs) can induce interleukin 1 (IL-1)-like activity. Oxidative stress that we used was superoxide anion (O2-) and hydrogen peroxide (H2O2). O2-, but not H2O2, could induce an IL-1-like factor(s) from monocytes and PMNs. IL-1-like activity from monocytes and PMNs induced by O2- was due to de novo synthesis because no IL-1 like activity was found in culture supernatants and in the lysate of unstimulated cells. We next examined the effects of radical scavengers on production of an IL 1-like factor(s). Generation of IL-1-like activity from monocytes was amplified by preincubation with catalase (H2O2 scavenger), although it was suppressed by preincubation with either superoxide dismutase (O2- scavenger) or vitamin E (antioxidant analogs). These results suggest that production of an IL-1-like factor(s) from monocytes and PMNs was due to O2- stimulation. Our data that production of an IL-1-like factor(s) from inflammatory cells by stimulation with O2- imply a model of the up-regulation mechanism of inflammation mediated by enhanced IL-1-like factor production stimulated with reactive oxygen species. PMID- 2553312 TI - [Evaluation of digital vibration threshold for asymptomatic ulnar neuropathy in patients with carpal tunnel syndrome]. AB - Quantitative assessment of vibration threshold at 125 Hz of index and little finger tips together with measurements of median and ulnar sensory nerve conduction velocity (SNCV) was performed for 25 limbs of 15 patients with carpal tunnel syndrome (CTS), 25 limbs of 13 disease controls with neurological symptoms similar to CTS and the nearly same number of limbs of normal controls. There was no significant difference in ulnar SNCV among 3 groups. However, ulnar SNCV at wrist was significantly reduced in patients with CTS as compared with normal value. In 17 limbs with CTS, vibration threshold of index finger tips was significantly increased, and 8 of those limbs were found to have concomitant increase of vibration threshold of little finger tips. Most of patients with increase of vibration threshold of both index and little finger tips were those who used to expose to repetitive mechanical stimuli at the wrist in their occupation. It was concluded that digital vibration threshold is simple to measure yet sensitive enough to detect asymptomatic ulnar neuropathy in CTS. PMID- 2553313 TI - [A case of MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes) with progressive cytochrome c oxidase deficiency]. AB - We report a 9 year-old boy with MELAS. High dosed oral thiamine administration and high fat diet induced remarkable neurological and biochemical improvement. His mother had episodic headaches and hemiplegia, probably MELAS. He complained muscle weakness and repeated episodes of vomiting started from 2 years of age. High levels of serum lactate and pyruvate were recognized, but with no metabolic acidosis. He developed generalized muscle weakness, growth retardation, generalized convulsions and stroke-like episodes at 5 years old. Optic nerve atrophy and mental retardation gradually appeared. A muscle biopsy at 5 years old revealed numerous ragged-red fibers with excess accumulation of lipid droplets and glycogen particles. Scattered fibers had no cytochrome c oxidase (CCO) activity representing focal CCO deficiency. An electron microscopy showed markedly increased number of giant mitochondria filled with markedly proliferated complicated cristae. Pyruvate dehydrogenase complex level in the fibroblasts was within normal ranges. Serum carnitine level was normal. With oral administration of thiamine hydrochloride (1000 mg) and high fat diet (60-70%), muscle weakness improved, and lactate and pyruvate levels in the serum reduced to normal ranges, whereas the mental deterioration, muscle atrophy, pes cavus progressed very slowly. He died from cardiac and renal failures at 9 years old. Autopsied muscles showed a marked decrease in cytochrome c oxidase activity (biochemically 12.8% of the normal level), and almost all muscle fibers had no cytochrome c oxidase activity histochemically. The progression of the MELAS was probably in parallel with the decrease in CCO activity. PMID- 2553314 TI - Serial dacryoscintigraphy before and after treatment with pseudoephedrine. AB - Radionuclide dacryoscintigraphy is an accurate physiologic method for evaluating the nasolacrimal drainage apparatus. The following case report illustrates the role of this procedure in the initial workup of epiphora, as well as its value in assessing response to therapy with pseudoephedrine. PMID- 2553315 TI - Analogy between tumor uptake of technetium(V)-99m dimercaptosuccinic acid (DMSA) and technetium-99m-MDP. AB - Sixty patients with a variety of malignant tumors were examined with Tc-99m(V) dimercaptosuccinic acid (DMSA) prepared by modification of a commercially available DMSA kit. Significant uptake of Tc-99m(V)-DMSA was observed in a number of tumors, offering additional clinically useful information. In the majority of cases in this study, however, the benefit of the Tc-99m(V)-DMSA image was limited because of low sensitivity. The most striking observation was the similarity between the tumor concentration of Tc-99m(V)-DMSA and the Tc-99m-MDP uptake in the tumor on the regular bone image. Therefore, patients with Tc-99m-MDP uptake in nonosseous tumor sites on the bone scan may be suitable candidates for tumor imaging with Tc-99m(V)-DMSA. PMID- 2553316 TI - Thallium-201 single photon emission computed tomograms of double cancers: lung and breast. AB - Thallium-201 SPECT is reported in a rare case with simultaneously developing double cancers of the lung and the breast. The TI-201 SPECT was initiated to evaluate mediastinal involvement from the lung cancer, and an abnormal tracer accumulation was unexpectedly found in the breast in addition to the pulmonary lesion. PMID- 2553317 TI - PYP Maalox localization in liver and spleen. PMID- 2553318 TI - Catecholaminergic and muscarinergic receptors in chronic experimental uremia. AB - In chronic uremic rats the density of alpha 1- and alpha 2-adrenoceptors in the cerebral cortex was significantly increased while beta-adrenoceptors and muscarinergic receptors in heart and brain as well as cardiac alpha 1 adrenoceptors were unchanged. Only the binding of the alpha 2-selective ligand 3H clonidine was inhibited by uremic serum. In isolated left ventricular muscle strips dose-effect curves of (-)-noradrenaline and (+/-)-dobutamine were not significantly different in uremic and control animals whereas the negative inotropic action of the muscarinergic agonist carbachol was attenuated. PMID- 2553320 TI - Correction of the deficient alveolar ridge. AB - The success of a denture is dependent on a good foundation; therefore, construction of the prosthesis begins with extraction of the teeth. The teeth should be maintained as long as possible if they can be restored. When the teeth are extracted, bone resorption usually starts, owing to systemic and local factors. Good communication between the prosthodontist and the oral and maxillofacial surgeon must exist during the diagnostic procedure, must be maintained through the various stages of treatment, and must prevail through the follow-up care of the patient. Each clinician must be aware of both the objectives and the possible limitations of the treatment the other will provide, to ensure optimal care for the edentulous patient. The development of the osteointegrated titanium implant system opens an exciting new avenue with respect to total rehabilitation of the edentulous patient. This is the future, and in our opinion, the preferred technique for mandibular reconstruction. PMID- 2553319 TI - Increased hexokinase/glucose-6-phosphatase ratio in the diabetic kidney as index of glucose overutilization. AB - In mice with streptozotocin-induced diabetes of 3 days' duration, the hexokinase/glucose-6-phosphatase (HK/G6Pase) ratio in the kidney was enhanced by 52% (mean +/- SEM: 0.40 +/- 0.04 vs. 0.26 +/- 0.03; p less than 0.02) compared to control mice as a result of a 25% increase of HK (16.68 +/- 0.93 vs. 13.31 +/- 1.04 nmol/min/mg protein; p = 0.05) and a 17% decrease of G6Pase (42.51 +/- 2.75 vs. 51.25 +/- 1.89; p less than 0.05). In contrast, as expected, the corresponding ratio (HK + glucokinase/G6Pase) was strikingly reduced in the liver. In 9-day diabetic mice, the kidney enzyme changes were much smaller; however, in a chronic disease such as diabetes, even minimal deviations from the normal may lead to significant metabolic changes with time. The enhanced HK/G6Pase ratio in the diabetic kidney suggests an increase in glucose utilization. This may contribute to the increased synthesis of glycogen, glycoproteins (including basement membrane) and RNA (via provision of ribose phosphate) occurring in the diabetic kidney and supports the view that the kidney (as opposed to other tissues) shows an 'anabolic response' to diabetes. PMID- 2553321 TI - Could bovine serum albumin be the initiating antigen ultimately responsible for the development of insulin dependent diabetes mellitus? AB - Western blotting of either a cloned rat beta-islet tumour cell extract or isolated BB rat islets with rat anti-bovine serum albumin antiserum revealed a cross-reacting protein (Mr = 69,000). A protein of similar molecular size was observed by fluorography in proteins immunoprecipitated from islet cells labelled with (35S)-methionine using anti-bovine serum albumin antiserum. In comparing the primary structure of the beta subunits of the proteins Ia, DQ and DR a region of homology with bovine serum albumin became evident. Analysis of the amino-acid homology in relation to the DR/DQ allotypes found in the human population gave a strong correlation between the combined DR and DQ homology score with bovine serum albumin and the incidence of insulin dependent diabetes mellitus. PMID- 2553322 TI - Glucagon storage, release and degradation by tumoral islet cells (RINm5F line). AB - Tumoral islet cells of the RINm5F line contain and release glucagon. Over 90 min incubation, the secreted hormone represents about 50% of the final cell content (1.4 +/- 03 fg/cell). L-arginine augments modestly glucagon release, the relative magnitude of such an effect being inversely related to the concentration of D glucose. The immunoreactive material secreted by the cells consists of both true glucagon (59-64%) and larger peptides. The RINm5F cells also release a proteolytic factor able to degrade exogenous glucagon or insulin. These results reveal that glucagon producing tumoral cells display several abnormal features in their secretory behaviour. PMID- 2553323 TI - Signal transduction by the platelet-derived growth factor receptor involves association of the receptor with cytoplasmic molecules. PMID- 2553324 TI - Inhibitory role of Ca2+ in the control of renin secretion: a study using superfused dispersed rat renal cortical cells. AB - 1. The role of Ca2+ in the control of renin release was investigated using a collagenase-dispersed rat kidney cortex cell preparation. 2. Superfusion with a series of low [Ca2+] buffers in either ascending or descending order of concentration increased renin release. Exposure to 0.06 mmol/l Ca2+ increased release by 120% (P less than 0.001) when presented as the first buffer in ascending order of concentration and by 79% (P less than 0.001) when presented as the fourth and last in a series of descending order. 3. The Ca2+ entry blocking drug diltiazem in a range of concentrations increased renin release and at 10(-5) mol/l diltiazem the mean stimulation was 35% (P less than 0.01). 4. 8-(N.N Diethylamino)octyl-3,4,5-trimethoxybenzoate (TMB-8) reduces the release of Ca2+ from intracellular stores and, studied over a range of concentrations, this compound increased renin release. At 10(-5) mol/l TMB-8 the mean increase was 44% (P less than 0.001). 5. None of these experimental manipulations, low [Ca2+], diltiazem or TMB-8, had any effect on the release of adenosine 3':5'-cyclic monophosphate into the cell superfusate, indicating that a decrease in intracellular [Ca2+] increases renin release by a mechanism which is independent of changes in adenosine 3':5'-cyclic monophosphate production. 6. Effects of low [Ca2+], diltiazem and TMB-8 on renin secretion were all shown to be reversible when superfusion with control buffer was resumed. PMID- 2553325 TI - Carbidopa does not affect the renal response to atrial natriuretic factor in man. AB - 1. The dependence of atrial natriuretic factor on renal dopamine for its renal effects in man was examined in 10 healthy volunteers using the dopa decarboxylase inhibitor carbidopa. 2. Each volunteer attended on two occasions, and received an infusion of atrial natriuretic factor (4 pmol min-1 kg-1) for 60 min after pretreatment with either placebo or carbidopa orally. These were administered in random, double-blind fashion. 3. A similar increase in plasma atrial natriuretic factor concentration was seen after atrial natriuretic factor infusion on both visits. 4. Infusion of atrial natriuretic factor produced a small unsustained rise in urinary dopamine excretion. This increase in urinary dopamine excretion was blocked by carbidopa with no effect on the natriuresis. 5. Urinary guanosine 3':5'-cyclic monophosphate excretion increased in response to the atrial natriuretic factor infusion whether placebo or carbidopa was given. Guanosine 3':5'-cyclic monophosphate, but not dopamine, may be a mediator of the renal response to atrial natriuretic factor in man. PMID- 2553326 TI - Metabolic changes in human liver associated with preoperative intravenous nutrition. AB - 1. This study examined the influence of pre-operative intravenous nutrition upon carbohydrate stores, glucose metabolism and protein synthesis in the liver of patients undergoing laparotomy. 2. Thirty patients with gastrointestinal cancer and weight loss (greater than 5 kg in 3 months) were randomized to receive a hospital diet only or a hospital diet plus intravenous nutrition (0.18 g of N + 125 kJ day-1 kg-1) for 3 or 7 days before laparotomy. Patients who had not lost weight received the hospital diet only and formed a control group. 3. Wedge biopsies of liver were obtained at laparotomy and analysed for glycogen concentration, the activity of three key enzymes of glucose metabolism, 6 phosphofructokinase (EC 2.7.1.11), fructose bisphosphatase (EC 3.1.3.11) and hexokinase (EC 2.7.1.1), and the capacity for protein synthesis. 4. Compared with controls and the hospital diet group, both phosphofructokinase and fructose bisphosphatase activity were reduced in patients who received intravenous nutrition, suggesting the utilization of glucose for glycogen synthesis with a reduction in the glycolytic flux. Consistent with these changes, patients who received intravenous nutrition had a significantly higher glycogen concentration compared with the control and hospital diet groups. 5. Maximal rates of protein synthesis were achieved after only 3 days of intravenous nutrition. 6. The provision of intravenous nutrition was associated with changes in hepatic metabolism suggestive of repletion of energy stores and a higher capacity for protein synthesis. PMID- 2553327 TI - Comparison between endogenous digoxin-like immunoreactivity and 86Rb uptake by erythrocytes in extracts of human plasma. AB - 1. To investigate endogenous cardiac glycoside-like compounds in plasma and their ability to inhibit the sodium pump, digoxin-like immunoreactivity [digoxin-like immunoreactive substance(s), DLIS] and 86Rb uptake by erythrocytes were measured in plasma extracts from normal adults, hypertensive adults and neonates. 2. DLIS levels in neonate plasma extracts were significantly higher than those found for normotensive or hypertensive adults. No difference was observed between normotensive and hypertensive subjects. DLIS was significantly increased when boiled plasma was extracted. 3. Extracts of boiled neonate and adult plasma inhibited 86Rb uptake. Instead, when boiling was omitted, no detectable inhibition was found in extracts of plasma from normotensive or hypertensive adult subjects. When present, the inhibition resulted from a depression of the ouabain-sensitive (sodium-pump-mediated) component, and, for the boiled neonate plasma only, also of the ouabain-resistant component. When the data from the different groups were pooled, a statistically significant inverse relationship between DLIS and erythrocyte 86Rb uptake was observed. Furthermore, in a subgroup of samples in which determinations were made before and after boiling in the same samples, an inverse correlation was found between changes in DLIS and changes in ouabain-sensitive (but not ouabain-resistant) 86Rb uptake. 4. Plasma extracts incubated with albumin at a physiological concentration significantly decreased (by approximately 20%) the inhibition of 86Rb uptake observed. 5. These findings support the existence of one or more endogenous compounds which both bind to antidigoxin antibodies and inhibit transmembrane cation transport. Part of this inhibition may, however, not involve the sodium pump. Furthermore, this chemically unidentified substance(s) may be bound to plasma proteins which partly reduce its action in vivo. PMID- 2553328 TI - Alterations in cerebrospinal fluid angiotensin II by sodium intake in patients with essential hypertension. AB - 1. Angiotensin (ANG) levels were measured in the cerebrospinal fluid of 15 patients with essential hypertension on a high sodium diet for 1 week and on a low sodium diet for a further week. ANGs were determined using a system of extraction by Sep-Pak cartridges followed by h.p.l.c. combined with radioimmunoassay. 2. Sodium depletion resulted in increases of ANG II in the cerebrospinal fluid from 1.16 +/- 0.38 (SEM) to 1.83 +/- 0.43 fmol/ml (P less than 0.01) and of ANG III from 0.65 +/- 0.11 to 0.86 +/- 0.15 fmol/ml (P less than 0.01). 3. The ANG II level in the cerebrospinal fluid was found to be unchanged and recovery of added ANG II was approximately 90%, even after incubation for 3 h, on both diets. Thus, it is unlikely that ANG II is produced or degraded in the cerebrospinal fluid in vitro. 4. There was no significant correlation between the cerebrospinal fluid and the plasma ANG II concentration on the low sodium diet. 5. These results suggest that the cerebrospinal fluid ANG II level increases with sodium depletion, and that the effect of the level of ANG II on the activity of the angiotensin-forming system in the central nervous system may be assessed by determination of ANG II in the cerebrospinal fluid in patients with essential hypertension. PMID- 2553329 TI - A comparative study of the enzymatic and toxic properties of venoms of the Asian lance-headed pit viper (Genus Trimeresurus). AB - 1. The lethalities, anticoagulant effects, hermorrhagic, thrombin-like enzyme, hyaluronidase, protease, arginine ester hydrolase, 5'-nucleotidase, L-amino acid oxidase, alkaline phosphomonoesterase, phosphodiesterase and phospholipase A activities of twenty-three samples of venoms from twelve species of Asian lance headed pit vipers (genus Trimeresurus) were examined. 2. The results indicate that notwithstanding individual variations in venom properties, the differences in biological properties of the Trimeresurus venoms can be used for the differentiation of venoms from different species of Trimeresurus. 3. The results also suggest that differences in the biological properties of snake venoms are useful parameters in the classification of snake species. 4. Our results indicate that venoms from the species T. okinavensis exhibited biological properties markedly different from other Trimeresurus venoms examined. This observation supports the recently proposed reclassification of T. okinavensis as a member of the genus Ovophis, rather than the genus Trimeresurus. PMID- 2553330 TI - Presence and properties of cAMP phosphodiesterase from promastigote forms of Leishmania tropica and Leishmania donovani. AB - 1. This paper reports the isolation and the partial purification of cAMP phosphodiesterase (EC 3.1.4.17) from the promastigote form of Leishmania tropica and a preliminary result from Leishmania donovani. 2. The activity of the fraction obtained from column chromatography was measured. 3. The effects of pH, temperature, time of incubation and various compounds on its activity in vitro were obtained. 4. Two peaks (I and II) exhibiting cyclic nucleotide phosphodiesterase activity were obtained. 5. Both activities were found to require the addition of Mg2+ ions for full effect. 6. The apparent Km values for the first and second peaks were 1.43 x 10(-3) M and 4.1 x 10(-3) M respectively. L. donovani shows only one peak of activity. PMID- 2553331 TI - Biochemical studies in marine species--I. NADP(+)-dependent isocitrate dehydrogenase from turbot liver (Scophthalmus maximus L.). AB - 1. An NADP+-dependent isocitrate dehydrogenase was extracted from turbot liver. The enzyme required divalent cations (Mg2+ or Mn2+) for its activity but was inhibited by high salt concentrations. 2. The enzyme had an optimum activity in the pH range between 7.5 and 9.0. The enzymic activity increased with temperature (in the range of 5 to 68 degrees C) with an Ea of 23.5 kJ/mol and a Q10 of 1.34. 3. The apparent Km values for the substrates were 6.5 microM for NADP+, 56 microM for Mg2+, 87 microM for Mn2+ and 4.2 and 73.5 microM for the complexes Mg isocitrate and Mn-isocitrate, respectively. The physiological significance of these results is discussed. 4. The enzyme was inhibited by citrate and adenine nucleotides. The degree of inhibition depended on the relation between the concentrations and that of magnesium. A possible regulating mechanism is proposed. PMID- 2553333 TI - Proteoglycan synthesis in vitamin D-deficient cartilage: recovery from vitamin D deficiency. AB - Vitamin D appears to be required for mineralization of skeletal elements. There is also evidence that cartilage proteoglycans may be involved in the regulation of mineralization. Previous studies have shown an alteration in the structure of the proteoglycans of the epiphyseal growth cartilage as a result of the decrease in serum calcium related to deficiency of dietary vitamin D. Vitamin D deficiency also induces a thickening of the epiphyseal growth plate presumably because of the inhibition of maturation of the growth plate chondrocytes. In order to compare the effect on proteoglycan structure with that on growth plate morphology, the proteoglycans of healing epiphyseal cartilage were characterized. The results indicate that, consistent with previous data, in vitamin D-deficient hatching chicks, the proteoglycans of the growth cartilage, but not of the articular cartilage, are smaller in monomer size with slightly smaller chondroitin sulfate chains whose sulfation pattern is unaltered. Sternal cartilage proteoglycans are unaffected. During recovery from vitamin D deficiency, the proteoglycans isolated from the growth cartilage are still not completely normal one day after supplementation with vitamin D, but are indistinguishable from normal by four days. In addition, the results conflict with those of a previous study in which only growth cartilage of hatchling chicks, not sternal or articular cartilage, was reported to synthesize large proteoglycans. Instead, all of these cartilages in the normal chicken have been found in this study to produce large proteoglycans of a size typical for mammalian cartilage and embryonic chick cartilage. PMID- 2553332 TI - Comparative subcellular distribution of apyrase from animal and plant sources. Characterization of microsomal apyrase. AB - 1. Apyrase (ATP: diphosphohydrolase) has been found in the microsomal fraction of rat salivary gland, mammary gland and uterus. 2. This enzyme, already described in plant tissue, is mainly present as a soluble polypeptide in tubers of Solanum tuberosum. 3. A fraction of this enzyme is associated with the microsomal fraction with a higher specific activity than the soluble one, for either ATP or ADP as substrate. 4. Apyrase bound to microsomes from rat and potato tissues was characterized in its substrate specificity and effect of inhibitors. 5. The Km values for ATP and ADP, optimum pH and metal ion requirement were determined. 6. A characteristic common to the microsomal and soluble apyrases is the stimulatory effect of a potato activator protein of soluble plant apyrase. 7. The microsomal bound apyrase from rat and potato tissues were solubilized and subjected to size exclusion chromatography. 8. The mammary gland and salivary gland apyrases eluted as molecular aggregates, in contrast to the uterus and potato enzyme. PMID- 2553334 TI - Effect of warm ischemic time on kidney transplants. PMID- 2553335 TI - A placebo-controlled, blind comparison of nedocromil sodium and beclomethasone dipropionate in bronchial asthma. AB - A multi-centre, randomized, blind comparative group study was carried out in 202 adult patients, who had suffered from asthma for at least 2 years, to assess the effectiveness and tolerability of maintenance treatment with either 4 mg nedocromil sodium 4-times daily, 0.1 mg beclomethasone dipropionate 4-times daily or 2 puffs of placebo 4-times daily, given by inhalation. Lung function (FEV1 and sRaw) measurements were made at the beginning and end of a 2-week baseline period and then after 3 and 6 weeks of treatment: assessment were also made of asthma severity. Patients recorded daily on diary cards details of morning and evening PEFR, usage of inhaled bronchodilators, severity of dyspnoea, cough and morning tightness. The results showed that, compared with placebo, both nedocromil sodium and beclomethasone dipropionate-treated patients showed an improvement in FEV1 and a reduction in sRaw values: PEFR increased slightly in all three groups. There was an improvement in asthma severity, diminished rate of dyspnoea and cough, and reduced usage of inhaled bronchodilators in patients receiving active treatment but not in those on placebo. Overall assessment of treatment efficacy by both investigators and patients showed that opinions were significantly in favour of active treatment over placebo. Treatment was well tolerated and no serious side-effects were reported. It was concluded that at the dosages used nedocromil sodium was comparable with and equivalent to inhaled beclomethasone dipropionate in nearly all of the parameters assessed, and both drugs were superior to placebo in the maintenance treatment of asthma in adult patients. PMID- 2553336 TI - Molecular mimicry: parasite evasion and host defense. PMID- 2553337 TI - Celiac disease: adenovirus and alpha gliadin. PMID- 2553338 TI - Studies of HIV infection and the development of Epstein-Barr virus-related B cell lymphomas following transfer of human lymphocytes to mice with severe combined immunodeficiency. AB - Mice with severe combined immunodeficiency (C.B-17 scid, hereafter SCID) accept xenografts of adult human peripheral blood leukocytes (PBL). The transplanted human PBL expand in number and survive for at least thirteen months and have been shown to reconstitute human immune function at both the T and B cell levels. Human immunoglobulin production is restored, and secondary antibody responses to antigens such as tetanus toxoid can be induced. All SCID mice reconstituted with 50 x 10(6) or more PBL from donors with evidence of exposure to Epstein-Barr virus (EBV) have developed human B cell lymphomas at 8-16 weeks after PBL engraftment, whereas mice reconstituted with PBL from EBV-seronegative donors fail to develop tumors. These tumors involve both lymphatic and non-lymphatic organs, and histologically they resemble large cell or immunoblastic lymphomas. The tumors are associated with high levels of human immunoglobulin secretion and serum electrophoresis reveals oligoclonal immunoglobulin banding patterns. Analysis of tumor DNA shows the presence of EBV genomes and oligoclonal patterns of immunoglobulin JH gene rearrangement. Taken together, these observations suggest an EBV-related proliferation of B lymphocytes leading to the rapid appearance of oligoclonal B cell malignancies following transfer of B lymphocytes from "normal" donors to SCID mice. SCID mice reconstituted with PBL from EBV seronegative donors have been infected with the LAV-1 strain of human immunodeficiency virus (HIV-1). Virus has been recovered from most infected animals by co-culture of mouse tissue with human T lymphoblasts. Some mice with high virus titers have developed an acute wasting syndrome and depletion of human T cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553339 TI - Telomeric associations in a lymphoblastoid cell line from a patient with B-cell follicular lymphoma. AB - We have established a new Epstein-Barr virus transformed cell line from a patient with B-cell follicular lymphoma. Telomeric fusions were observed in several subclones, with the nonrandom involvement of chromosomes 1, 5, 12, and 17. Centromeric staining with immunofluorescent anti-kinetochore antibodies was positive in both centromeres of the fused chromosomes, suggesting they were both active. Unlike previously reported cases, we were unable to demonstrate telomeric fusions directly in cells from the patient's blood. However, the finding of identical immunoglobulin gene rearrangements in DNA from the patient's blood and cell line suggested that they originated from the same malignant B-cell clone. PMID- 2553340 TI - Experience with 7,604 bone scintigraphies at time of operation for breast cancer 1977-1987. AB - From 1977 to 1987, 7,604 out of a total of 20,483 Danish breast cancer patients had a bone scintigraphy in order to exclude or localise osseous metastases. The scintigrams were performed and interpreted at 14 local hospitals. In 1979, standardised guidelines for interpretation were agreed upon. Until 1983, the scintigrams were described centrally, based on the local reports. From 1979, the frequency of positive bone scintigrams suggesting bone metastases stabilised at about 5% of the patients compared with 12 and 20% in 1977-78. The local take-over of description did not change the frequency. The frequency was also stable from 1979 when evaluated in age groups. With increasing age the frequency of positive scintigrams increased. The frequency of positive scintigrams was significantly lower in patients entering a protocol than in those not entering a protocol. The conclusion is that on a nation-wide basis it is possible to establish a stable, uniform interpretation of bone scintigrams after a two-year introduction period. PMID- 2553341 TI - Pulmonary disease in patients with human immunodeficiency virus infection. AB - Pulmonary disease is the most important cause of morbidity and mortality in patients infected with human immunodeficiency virus (HIV). All parts of the hospital system are expected to be involved in the diagnosis and treatment of HIV infected patients in the coming years. Many different processes cause pulmonary disease alone or in combination. Bilateral interstitial infiltrates are the most frequent chest x-ray abnormality and are most frequently caused by infection with Pneumocystis carinii. Cytomegalovirus, Mycobacterium tuberculosis, nonspecific interstitial pneumonitis and pulmonary Kaposi's sarcoma are the most important parts of the differential diagnosis. An aggressive approach to the diagnosis of pulmonary disease in this patient population is indicated in order to provide optimal care and assess new therapies. PMID- 2553342 TI - DNA flow cytometric studies of 66 human lung tumors analyzed before treatment. Prognostic implications. AB - To investigate the prognostic implications of DNA flow cytometry in human lung tumors, we analyzed specimens from patients with neoplastic and non-neoplastic lung disease. Most non-neoplastic and normal (taken at the resection border) lung samples yielded a single cell population with diploid DNA content (only two normal lung specimens from two cancer patients had aneuploid DNA content). At least one aneuploid cell subpopulation was seen in 91 percent of NSCLC and 50 percent on SCLC. To show intratumor heterogeneity, multiple-site sampling was done whenever possible in both primary tumor and metastatic sites, revealing a high incidence of multiclonality (50 percent). Although diploid tumors were rare, they associated with a higher survival rate than aneuploid monoclonal and multiclonal tumors with hypoploid and/or hypertetraploid clones, which had the lowest survival. Cellular DNA content analysis in patients with lung tumors may be useful in prognostic evaluation. PMID- 2553343 TI - Preoperative examination to detect distant metastasis is not advocated for asymptomatic patients with stages 1 and 2 non-small cell lung cancer. Preoperative examination for lung cancer. AB - The purpose of the present study was to determine whether all patients with non small cell lung cancer, clinically defined as stage 1 or 2, should have examinations to determine the presence of distant metastatic (M1) disease. The survival rates of patients who underwent the examinations and those who did not were compared. In all groups, the examinations had in no way positively affected survival. The 33 patients in whom distant recurrence had occurred within 12 months of curative operation, were then evaluated to establish the relationship between the recurrent site and the preoperative examination: negative scans on the examination failed to predict the low incidence of early distant recurrence. Those data suggest that the routine use of radionuclide or CT scans is of no benefit to asymptomatic patients with local, early disease. PMID- 2553344 TI - Lung collagenase inhibitors and spontaneous and latent collagenase activity in idiopathic pulmonary fibrosis and hypersensitivity pneumonitis. AB - In order to analyze the mechanisms involved in the decreased collagenolytic activity previously observed in interstitial lung fibrosis, we studied the inhibitory collagenase activity and the latent activable collagenase in lung samples from five patients with IPF, six with HP, and three control subjects. Our results showed that in both diseases, the inhibitor levels were significantly higher than in control subjects. Findings suggest that in IPF low amounts of collagenase plus excessive enzyme-inhibitors may be operating to decrease collagen catabolism. In contrast, HP lungs seem to contain adequate amounts of the enzyme but higher levels of inhibitors play a role in the abnormal degradation observed in some patients. PMID- 2553345 TI - Airway hyperresponsiveness. Mechanisms in experimental models. PMID- 2553347 TI - Hypercalcemia in atypical bronchial carcinoid tumors. AB - Atypical carcinoid tumors of the bronchial tree are uncommon. Their tendency to metastasize is well recognized, characteristically producing osteoblastic bone deposits without disturbance of calcium homeostasis. We report two patients who presented with hypercalcemia and osteolytic bone metastases following surgical removal of atypical bronchial carcinoid tumors. In one of the patients, chemotherapy induced remission and controlled the hypercalcemia. PMID- 2553346 TI - Phrenic involvement in Charcot-Marie-Tooth disease. A pathologic documentation. AB - A 69-year-old woman diagnosed as having Charcot-Marie-Tooth disease (CMT) for 30 years' duration died of respiratory failure secondary to diaphragmatic weakness. At autopsy, identical neuropathic changes were found in phrenic and other affected somatic peripheral nerves. We conclude that the phrenic nerve may be involved by the neuropathy of at least some forms of CMT. PMID- 2553348 TI - [Experiences with the endorectal ileal pull-through operation with lateral ileal reservoir following colectomy in ulcerative colitis and colonic polyps]. PMID- 2553349 TI - [The direct ileum pouch-anal anastomosis. Clinical and functional continence results]. PMID- 2553350 TI - [Longitudinal myotomy as an alternative to conventional pouch procedures following proctocolectomy]. PMID- 2553351 TI - [A study on cytomegalovirus infection in 199 pregnant women and their infants]. AB - Cytomegalovirus (CMV) infection is common in human, and exerts harmful effects during pregnancy upon fetuses. In order to get a clear knowledge of CMV infection rate in pregnant women and their infants and of the relationship between maternal infection and congenital infection in our city, we tried to detect the CMV specific IgG and IgM in 199 paired serum samples of mother and infant. The results showed that 183 maternal serum samples and 179 umbilical cord serum samples were CMV-IgG positive, a positive rate of 92% and 90% respectively. While 6 maternal serum samples and 7 umbilical cord serum sample were CMV-IgM positive, a positive rate of 3.0% and 3.5% respectively. The close correlation of CMV-IgM levels between mothers and their babies indicates that the infant may acquire congenital infection from his CMV-IgM positive mother. PMID- 2553352 TI - [Carrier status of herpes viridae in pregnant women]. AB - 282 cervical swab specimens and serum samples collected from pregnant women were used for virological studies in order to investigate the carrier status in herpes viridae and antibody. Seven isolates were obtained with a positive rate of 2.48%. According to the characteristics of cytopathology effects on the diploid cells (2BS), pathogenicity in the hosts and electron micrographic findings, these 7 isolates belonged to herpes viridae. The results of neutralization test, enzyme linked immunosorbent assay blocking test and indirect immunofluorescence tests against HCMV-specific serum and standard HCMV AD-169, HSV-ISm44, HSV-IIG indicated that all these 7 viruses were HCMV. No other viral members of herpes viridae were detected in this study. PMID- 2553353 TI - [Evaluation of antipyrine clearance in chronic liver diseases]. AB - Antipyrine (AP) clearance was determined in 23 cases with liver cirrhosis (LC), 12 with chronic active hepatitis (CAH), 12 with hepatocellular carcinoma (mcHCC), 20 with non-hepatic diseases and 70 healthy controls. ICG Clearance was performed simultaneously in 9 cases of them. The results showed that AP clearance was significantly decreased in patients with LC and moderately decreased in CAH and HCC, its diagnostic sensitivity in LC was significantly higher than that of GPT. The significant positive correlation between the AP and ICG clearance was noted and AP clearance also well correlated with serum albumin level and prothrombin time. It is suggested that AP clearance may be used as a quantitative test to determine the reserve capacity of liver and as a substitutive test for ICG clearance. PMID- 2553355 TI - [Small cell lung cancer with Lambert-Eaton myasthenic syndrome]. AB - Four of 69 cases of small cell lung cancer (SCLC) showed evidence of Lambert Eaton myasthenic Syndrome (LEMS) were studied neurologically and neurophysiologically in four years. The LEMS appearance were preceded that of SCLC in 3 cases for two years at most. Repeated stimulation of ulnar nerve examination showed diminished amplitude of initial response (0.2-0.9 mv); Amplitude of the response at 3 c/s stimulation for 3 sec was diminished 20-63%. (control 3% decreases-13% increases) but that at 20 c/s stimulation for 10 sec increased 200-800% increases. (control: 20% decreases-56% increases). These findings were important for diagnosis of LEMS. The abnormalities of neurotransmission seems to be due to inadequate release of acetyl choline from nerve terminals at abnormal active zone of Ca++ channels. PMID- 2553354 TI - Identification and regulation of the eukaryotic hyaluronate synthase. AB - Hyaluronate synthesis is required for fibroblast detachment in mitosis and migration. It is regulated by the activity of the synthase which is localized at the inner side of plasma membranes. The synthase was identified as a 50 kDa protein by immunological cross-reaction with the streptococcal enzyme and by affinity labelling. Transformation of fibroblasts by Rous sarcoma virus activated the synthase by enhanced transcription and phosphorylation. The synthase was a natural target of pp60v-src kinase. PMID- 2553356 TI - [Uremic neuropathy]. AB - Two autopsy cases of uremic neuropathy were reported. The chief clinical features were progressive peripheral sensory impairment, especially of the deep sensation, and the distal weakness of the four extremities. Histopathological findings revealed not only marked segmental demyelination and axonal degeneration of the peripheral nerves, but also severe damage of the skeletal muscles, including the necrosis of the myofibers, streaming of the Z-band, aggregation of glycogen, and the presence of membranous bodies under the electron microscope. It indicated that both the peripheral nerves and the skeletal muscles were involved in some metabolic diseases. PMID- 2553357 TI - [Factors influencing long-term prognosis after pancreatoduodenectomy in cancer of the pancreas head]. AB - Sixty-four cases of pancreatoduodenectomy were performed for ductal adenocarcinoma of the head of pancreas. According to the size of primary tumor, the cases were divided into 3 groups: less than 2 cm, 2.1 to 4.0 cm, and more than 4 cm. The corresponding one-year survival was 66.7%, 33.7%, and 25.0% (P less than 0.05), respectively. The one-year survival of cases with tumor confined to pancreas or only involving intrapancreatic common bile duct or duodenal wall in the ampullary region (100%) was higher than that of cases with microscopic and macroscopic local infiltration (33.3%, 54.5%) and that of cases with large or middle extrapancreatic vessels involved (11.1%) (P less than 0.001). The rate of lymph node metastases of all cases was 69%. The one-year survival of cases with primary lymph node metastases was lower than that of cases without it (32.4% v. 62.5%, P less than 0.05). Based on the above mentioned result, a new staging method is suggested. PMID- 2553358 TI - [Surgical management of tracheal tumors]. AB - Eighteen operations on 16 patients with tracheal tumors were reported. Of these cases, 11 had primary tracheal tumors and 6 had secondary ones. The tumors were malignant in 10, low malignancy in 3, and benign in the remaining 3 cases. The diagnostic procedure, operative technique, and management of postoperative complications are discussed. PMID- 2553359 TI - Serum beta 2-microglobulin before and after renal and heart allotransplantation and in infection. AB - beta 2-microglobulin (beta 2-m) serum levels were significantly elevated in patients waiting for kidney transplantation. The levels decreased after successful transplantation, even when they were never as low as in a control group of healthy blood donors. A higher amount of serum (s-) beta 2-m was observed also in heart transplant recipients. A significant reincrease in s-beta 2-m occurred during rejection of renal allograft and during CMV and EBV infection. The levels of s-beta 2-m were higher in patients infected with CMV than in those infected with EBV. ABBREVIATIONS: beta 2-m = beta 2-microglobulin, s-beta 2-m = serum beta 2-microglobulin, CMV = cytomegalovirus, EBV = Epstein Barr virus. PMID- 2553360 TI - [Cholesterol content and malignant proliferation of cancer cells]. AB - The relationship between cholesterol content and proliferation of cancer cells was studied. When the cholesterol content of cancer cells increased. cAMP content of the cells also increased; the agglutinability of the cells decreased; the percentage of diploid cells increased greatly (from 54% to 79%) and the tetraploid cells reduced rapidly (from 43% to 15%). It was shown that the malignant proliferation of cancer cells decreased obviously. The cholesterol was not absorbed markedly by the V79 cell line and embryo lung cells as compared with the cancer cells when cholesterol content in the medium was increased. There were no apparent changes in the percentages of diploid cells or tetraploid cells. PMID- 2553361 TI - [Correlation of human papilloma virus (HPV) and herpes simplex virus-2 (HSV-2)in the viral etiology of carcinoma of uterine cervix]. AB - HPV DNA and HSV-2 antigens were detected in a series of cervical lesions by DNA hybridization and PAP stain techniques in order to investigate the possible role of HPV and HSV-2 in the viral etiology of cervical carcinoma. The correlation of HPV DNA sequence and HSV-2 antigens was analyzed. The results showed that the positive rates of HPV 16, 18 and 11 in cervicitis, dysplasia and cervical cancer were 13%; 0, 13%; 17%, 17%, 17%; 43%, 8%, 4%; and HSV-2 antigens were found in 41% of cervicitis, 50% of cervical dysplasia and 63% of cervical carcinoma. Comparing the results of HPV hybridization with that of HSV-2 antigens, the authors found that both HPV DNA and HSV-2 antigens could be detected in some cases. The results indicate that the viral etiology of cervical carcinoma may be multifactorial and HSV-2 and HPV may be synergistic in the development of cervical carcinoma. PMID- 2553362 TI - [Distribution and ultrastructural localization of carcino-embryonic antigen (CEA) in signet-ring cells of gastric cancer]. AB - The distribution and ultrastructural localization of CEA in signet-ring cells of 15 gastric cancer specimens were observed by PAP and immunoelectron microscopic methods. The mechanism of abnormal distribution of CEA in the signet-ring cell and its biological significance are discussed. The results showed that the CEA positive rate in signet-ring cells was 100% with the polarity lost in distribution. Under the light microscope, the CEA stain patterns were of two types-cytoplasmic and membranous types. The former was predominant. Under the electron microscope, most of the CEA was distributed on the cell membrane and cytoplasm. CEA was found in intracellular membranous structure of the cancer cells, especially in protein synthesis and transport organellae (RER, Golgi Complex etc). The synthesis of CEA in cancer cells increased, yet its elimination was somewhat hampered. The result was that the RER became extended and were full of CEA (+) material. In the free signet-ring cell, there was a small and short contact plane. The tight junction was severed as the cell junction was reduced. The antigenic determinant of CEA was glycoprotein. The abnormal distribution of CEA in signet-ring cells might be the morphologic reflection of the glycosylation of surface glycoprotein of tumor cells. These abnormal changes would lead to mal functions and biologic misbehavior in the cells. It may even lead to disturbances in connection and recognition, loss of contact inhibition and decrease in the adhesion between tumor cells and therefore may easily give rise to infiltration and metastasis. PMID- 2553363 TI - [Relationship between cyclic nucleotide phosphodiesterases (cPDE) and some patho biologic behaviors of stomach cancer--I. Histochemical studies of CPDE in stomach cancer tissues]. AB - Distributions and activities of both cyclic adenosine 3',5'-monophosphate phosphodiesterase and cyclic guanosine 3',5'-monophosphate phosphodiesterase (cAMP-PDE and cGMP-PDE) in 43 human stomach cancer tissues were studied histochemically. The relationship between cyclic nucleotide phosphodiesterases and some patho-biologic behaviors of stomach cancer was preliminarily discussed, and so was the application of these two enzymes in clinical practice of gastric cancers. The Results showed that the two cyclic nucleotide phosphodiesterases were closely correlated with some patho-biologic behaviors of stomach cancer. The authors considered that the histochemical investigation of cyclic nucleotide phosphodiesterases in stomach cancer tissues can be useful in determining the malignant degree and estimating the prognosis of stomach cancer objectively. PMID- 2553365 TI - [Malignant tumor with peripheral neuropathy as presenting symptom--clinical analysis of 52 cases]. AB - Fifty-two cases of malignant tumors with peripheral neuropathy as presenting symptom are reported. The duration of the peripheral neuropathy varied from 2 to 11 months before the malignant tumors were confirmed. The initial peripheral neuropathy included cranial nerve, sciatic nerve, intercostal nerve, brachial plexus, multiple peripheral nerves, thoracic outlet syndrome, polyneuritis, peripheral neuritis, autonomic nerve and Horner's syndrome. Most of these malignant tumors were lung cancer, nasopharyngeal cancer and hepatic carcinoma in order of incidence. Pathogenesis, clinical features, diagnosis and treatment of these tumors are discussed with review of the literature. The author emphasizes that if there was peripheral neuropathy of unknown cause, examination has to be done in detail and repeatedly for early diagnosis and better result. PMID- 2553364 TI - [Combined UFTM for 140 patients with advanced gastric cancer]. AB - 140 patients with advanced gastric cancer confirmed by pathology were treated by UFT (Uracil, FT-207) and mitomycin C (MMC) from Sept. 1985 to June 1987. All the patients received UFT #3 T. i. d. to a total dose of 30 g. Mitomycin C, 8-20 mg i. v. Q. wk was given to a total dose of 48-60 mg. Of the 140 patients, 65 had cancer of cardia, 36 cancer of gastric body, and 39 cancer of gastric antrum. There were 125 males and 15 females. The ages ranged from 30 to 80 years. In this series, CR was 10.0% and PR 44.3% with total remission rate of 54.3% (76/140). Thirty-four patients receiving UFT alone as control had a response rate of 26.5%. The response rate was higher in patients treated with UFTM than those with UFT alone. The median remission was 4 months. That main side effects were leukopenia and thrombocytopenia. The results showed that combination chemotherapy (UFTM) was valid in treating advanced gastric cancer. PMID- 2553366 TI - [Hepatic artery embolization for primary hepatic carcinoma]. AB - Twenty patients with primary hepatic carcinoma (PHC) treated by hepatic arterial embolization in our department from Dec. 1986 to Mar. 1987 are reported. There were 15 males and 5 females. The ages ranged from 34 to 75 years with an average of 50.7. Preoperative diagnosis and localization of the tumor were done by AFP, B us, CT and angiography (right lobe 15 cases, left lobe 1 case, both lobes 4 cases). Celiac and superior mesenteric angiography was carried out by femoral artery approach and then highly selective hepatic catheterization was utilized for hepatic arterial embolization. Antitumor agent (5-Fu, adriamycin), iophendylate and foamy gel sponge were used for peripheral and proximal embolization. Manifestations were improved in most of the patients after embolization, such as relief of abdominal pain, improvement of appetite, decrease of tumor size. Total necrosis of the tumor was found in 2 patients who underwent surgery 1 month after embolization. The side effects of the posthepatic embolization such as, nausea, vomiting, abdominal pain and fever could be relieved by symptomatic treatment. No severe complications, such as gangrene of the gall bladder, hepatic failure, liver abscess, intestinal necrosis or pulmonary embolization were found except 3 patients who died of renal failure after the procedure. The liver dys-function returned to normal within 2 weeks. Hepatic arterial embolization provides an alternative treatment for the patients with PHC who has compensated liver function without severe systemic diseases, especially renal endocrine problems and severe portal hypertension. They should have patent portal system as proved by angiography. The authors considered that this therapeutic embolization with hepatic chemotherapy infusion is safe and effective in the management of PHC. It may increase the resectability and provide palliative means for the advanced and terminal cases. PMID- 2553367 TI - Regulation of biological functions by insulin and insulin-like growth factor I via their own distinct receptors. PMID- 2553368 TI - [Papilloma viruses in neoplasms of the female sex organs]. PMID- 2553369 TI - [The mechanism of membranotropic action of 1-ethoxysilatrane]. PMID- 2553370 TI - [The effect of amidinomercaptic acids on prolyl endopeptidase and carboxycathepsin activity (angiotensin converting enzyme)]. PMID- 2553371 TI - [Topoisomerase I cleavage site in the regulatory zone of transcriptionally active minichromosomes of the SV40 virus located adjacent to a region, hypersensitive to DNAase I and segments recognizing topoisomerase II]. PMID- 2553372 TI - [Analysis of signal transmission in interneuronal synapses based on the reflection principle]. PMID- 2553373 TI - [Neural regulation of endocrine gland functions upon aging]. PMID- 2553374 TI - [Noradrenaline regulates the degree of methylation of DNA in the brain]. PMID- 2553375 TI - [The population of cyclic GMP-specific phosphodiesterases in bovine retina rod outer segments differing in affinity to photoreceptor membranes]. PMID- 2553376 TI - [Glutamate receptors in the human brain visualized using monoclonal antibodies]. PMID- 2553377 TI - [Synthesis and secretion of a hybrid polypeptide containing bull adrenocorticotrophic hormone, by Escherichia coli cells]. PMID- 2553378 TI - [Accumulation of cyclic AMP in frog urinary bladder tissue determines the gradual character of increase in osmotic permeability stimulated by vasopressin]. PMID- 2553379 TI - Regulatory mechanism of gene expression of methylcholanthrene-inducible cytochrome P-450. PMID- 2553380 TI - [Determining the prognosis in bronchial carcinoma by intraoperative pleural lavage]. AB - Pleural lavage with 300 ml physiological saline was undertaken at thoracotomy (before and after resection) in a prospective study of 59 patients (49 males and 10 females; average age 59.7 years, range 41-76 years) with first manifestations of a bronchial carcinoma. The aim was to find out whether, even at an early stage of the tumour, there are tumour deposits in the pleural cavity. The lavage fluid was centrifuged and the sediment then embedded in paraffin and prepared for cytological examination. Tumour cells were demonstrated before resection in 27 patients (45.8%), in 23 of them also after the resection. Among 38 patients, previously classified as being in stage I, the lavage material was positive in 15 (39.5%); tumour cells were demonstrated in 5 of 18 patients in stage pT1. The cumulative two-year survival rate of non-small-cell bronchial carcinoma in stage I (n = 32) was 39.9% if the lavage was positive, 97.4% if negative (P less than 0.01). The results indicate that a positive finding on pleural lavage corresponds to a pT4 stage with a poor life expectancy. PMID- 2553381 TI - [Disseminated polyneuropathy after coma]. AB - An acute polyneuropathy developed during intensive-care treatment of three young men who had sustained severe multiple traumas and of one woman with bacterial meningoencephalitis. In the first case there was a predominantly distal paraparesis of the legs; in the second, flaccid neurological deficits occurred particularly in the areas of the left ulnar, peroneal and tibial nerves combined with paralysis of cranial nerves IX, X and XII. A symmetrical sensorimotor polyneuropathy with tetraplegia and cranial nerve deficits occurred in the third case, and an asymmetrical sensorimotor polyneuropathy in the female patient. In all four patients electrophysiological tests demonstrated the pattern of neurogenic damage. All patients had respiratory failure, malnutrition, septic fevers and acute renal failure requiring haemodialysis for two to five weeks. It is assumed that the polyneuropathy was of multifactorial genesis. PMID- 2553382 TI - [Bronchial carcinoid. A clinical study of 37 patients]. AB - To establish whether bronchoplastic procedures designed to minimize loss of lung tissue are justifiable for the treatment of bronchial carcinoid tumours, data were analysed from 37 patients (17 men, 20 women, average age 51 [22-70] years) who had undergone surgery for typical (n = 30) or atypical (n = 7) bronchial carcinoids. Conventional tumour resections had been performed in 29 cases and bronchoplastic operations in eight. After an average observation period of 54 months one patient who had undergone lobectomy for a bronchial carcinoid had died of recurrent tumour, and one other patient who had been treated by pneumonectomy for an atypical carcinoid had developed distant metastases. All the other patients were free from tumour at that time. This indicates that patients treated by bronchoplastic procedures do not have any higher incidence of recurrences or any lower chance of survival than those treated by lobectomy or pneumonectomy. A bronchoplastic operation should therefore be the treatment of first choice, provided that the adjacent lung tissue has not been destroyed by retention pneumonia and that lymph node dissection does not reveal any involvement. PMID- 2553383 TI - [Malignant somatostatinoma of the papilla major and minor]. AB - In a 50-year-old man with extrahepatic cholestasis and iron-deficiency anaemia, local operative excision of a 4 cm submucous tumour of the papilla of Vater revealed on histological and immunochemical examination a somatostatinoma. Subsequent duodenopancreatectomy further demonstrated in the surgical specimen a lymph node metastasis and a 5 mm somatostatinoma in the papilla minor. The patient has now been free of symptoms for 12 months on substitution treatment with pancreatic enzymes. PMID- 2553384 TI - Axolotl pronephric duct cell migration is sensitive to phosphatidylinositol specific phospholipase C. AB - On the basis of its distribution pattern in embryos of the axolotl (Ambystoma mexicanum), we recently identified alkaline phosphatase as a molecule potentially involved in guiding the migration of the pronephric duct. Alkaline phosphatase is a cell surface protein anchored to cell membranes via a covalent linkage to a phosphatidylinositol glycan (PI-G). The enzyme phosphatidylinositol-specific phospholipase C (PIPLC) specifically releases from cell surfaces molecules anchored by the PI-G linkage. In order to test the possibility that a PI-G anchored protein is involved in directing pronephric duct cell migration, PIPLC was applied to axolotl embryos. The enzyme was introduced into embryos through the use of a novel slow-release bead material, hydrolysed polyacrylamide. PIPLC blocked pronephric duct cell migration without interfering with somite fissure formation, a concurrent, neighbouring morphogenetic cell rearrangement which occurs with little if any alkaline phosphatase present. In addition, alkaline phosphatase activity was markedly diminished in the vicinity of the implanted beads. These observations suggest that at least one protein anchored to the cell membrane by a PI-G linkage, possibly alkaline phosphatase, is involved in guiding or promoting pronephric duct cell migration. PMID- 2553385 TI - Mechanism of erythropoietin action on the erythroid progenitor cells induced from murine erythroleukemia cells (TSA8). AB - Erythropoietin is a well-known erythroid differentiation and growth factor, but the mechanism of its action is not well understood. In this work, we have examined its mechanism of action on the erythropoietin-responsive murine erythroleukemia cells (TSA8). TSA8 cells become responsive to erythropoietin after induction with DMSO. Stimulatory effects on erythropoietin response are observed with the addition of compounds affecting the cAMP level such as forskolin, phosphodiesterase inhibitor and cholera toxin only in the presence of erythropoietin. cAMP analogues themselves show no stimulatory effect on TSA8 cells, nor does erythropoietin increase cAMP level in the cells. Thus, it is suggested that cAMP does not act as a direct second messenger for signal transduction through erythropoietin receptors, but as a stimulator of the erythropoietin receptor pathway and/or as a second messenger in combination with the receptor pathway. The mechanism for acquisition of responsiveness to growth and differentiation factors of progenitor cells is discussed. PMID- 2553386 TI - Cyclic-AMP-induced elevation of intracellular pH precedes, but does not mediate, the induction of prespore differentiation in Dictyostelium discoideum. AB - Prespore gene expression in Dictyostelium is induced by the interaction of cAMP with cell surface cAMP receptors. We investigated whether intracellular pH (pHi) changes mediate the induction of prespore gene expression by cAMP. It was found that cAMP induces a 0.15 unit increase in pHi within 45 min after stimulation. The cAMP-induced pHi increase precedes the induction of prespore gene expression, measured by in vitro transcription, by about 15-30 min. Cyclic-AMP-induced pHi changes can be bypassed or clamped by addition of, respectively, the weak base methylamine, which increases pHi, or the weak acid 5,5-dimethyl-2,4 oxazolidinedione (DMO), which decreases pHi. Bypass of the cAMP-induced increase of pHi with methylamine does not induce the expression of prespore genes, while inhibition of the pHi increase with DMO does not inhibit the induction of prespore gene expression. Cyclic-AMP-induced prespore protein synthesis and the proportion of prespore cells in multicellular aggregates are also not affected by bypassing or inhibiting the cAMP-induced pHi increase. These results show that although a morphogen-induced pHi increase precedes the induction of prespore gene expression, this increase does not mediate the effects of the extracellular cAMP signal on the transcription or translation of prespore genes in Dictyostelium discoideum. PMID- 2553387 TI - Developmental profile of kappa, mu and delta opioid receptors in the rat and guinea pig cerebellum. AB - The phenotypic expression of the three opioid receptors kappa, mu and delta was determined in the rat and guinea pig cerebellum during ontogeny. In both species, the increase in cerebellum weight was accompanied with a gradual increase in the total number of opioid receptors. At the same time, however, the receptor concentration (femtomoles per milligram protein) fluctuated, and finally declined by 3.3- and 2.0-fold in the adult rat and guinea pig cerebella, respectively. In the guinea pig, kappa receptors constituted about 80% of the receptors at the earliest developmental stage tested, day 20 of the embryo, but then there was a sharp decrease in this value, resulting from an increase in both mu and delta receptors. This decrease was, however, transient, and the percentage of the kappa receptors increased again to reach 77% of the receptors in the adult cerebella. A similar transient increase in the percentage of mu and delta receptors was observed during the development of the rat cerebellum, although the receptor density in this species was 1/4-1/8 that of the guinea pig. Cultures of aggregating guinea pig cerebellar cells were used to further study the factors regulating the expression of the three opioid receptors in this brain region. The possibility that the development in vivo of cells expressing different opioid receptors is associated with the ontogeny of the endogenous opioid peptides is discussed. PMID- 2553388 TI - [Problems in the management of drug-resistant herpesvirus infections]. PMID- 2553389 TI - [Endothelin]. PMID- 2553390 TI - [Sulfonylurea receptors]. PMID- 2553391 TI - [Study of the effect of sclareol glycol diterpene on the 3',5'-AMP level]. AB - Sclareol glycol (SG) is a semisynthetic diterpene of the labdanic group. SG is tetranorlabdaniol bivalent alcohol 13, 14, 15, 16-tetranorlabdan-8 alpha, 12 diol, obtained from the plant Salvia sclarea L., grown in Bulgaria. The effects of SG were studied on the level of 3',5'-AMP in:mono-layer tissue cultures of the anterior hypophysis of the rat; in hypophysial and brain tissue during in vitro incubation; in liver microsomes of the rat during in vitro incubation and in liver perfusate of the rat. It was established that SG induced an increase in accumulation of 3,5'-AMP in the tissues from the anterior hypophyses of mature and newborn rats, in the sections of brain cortex and cerebellum of mature rats. Similar changes were found by the diterpene forskolin, studied comparatively. SG induced a reduction of accumulation of 3',5'-AMP in liver microsomes and diminished the release of 3',5'-AMP in a liver perfusate of the rat. The increased accumulation of 3',5'-AMP due to SG could be explained by postreceptor stimulation of the enzyme adenylate cyclase, binding directly to its catalytic subunit. The reduced accumulation of 3',5'-AMP in liver microsomes and its diminished release in liver perfusate could be explained either by activation of the inhibitory adenylate cyclase in hepatocytes or by desensitisation of 3',5' AMP-generating system. PMID- 2553392 TI - [Study of the effect of sclareol glycol diterpene on the release of adenohypophysial hormones prolactin, somatotropin and adenocorticotrophic hormone]. AB - The effects of a semisynthetic diterpene sclareol glycol (SG) on the release of adenohypophysial hormones prolactin, somatotrophic hormone (STH) and adrenocorticotrophic hormone (ACTH) were studied in the plasma of the rat. The effects of SG were investigated also parallel with diterpene forskolin (F) on the release of prolactin and STH in tissue cultures of adenohypophysial cells of the rat. The experiments established that SG increased in a low dose, but in higher dose lowered the concentration of plasma prolactin and STH. These effects were found even after 15 minutes after its intraperitoneal administration. SG induced mainly an increase of plasma concentration of ACTH. In tissue cultures of adenohypophysial cells SG caused lowering of prolactin concentration and an increase of STH; under these conditions F caused only an increase in the concentration of these hormones. The results show that SG influences secretion of prolactin, STH and ACTH in various ways in comparison with F. This might be due to th their various chemical structure and to their basic mechanisms of action (mainly by 3',5'-AMP for P; besides 3, 5'-AMP substantial participation of DA ergic and GABA-ergic mediation for SG). PMID- 2553393 TI - The distribution of amosite asbestos fibers in the lungs of workers with mesothelioma or carcinoma. AB - We have previously shown that there are differences in the sizes of fibers of amosite asbestos in different parts of the lung in workers with relatively high asbestos exposure and malignant pleural mesothelioma. To determine whether this distribution pattern is specific to cases of mesothelioma, we compared the fiber distribution in the lungs of 20 cases of mesothelioma and 10 cases of carcinoma of the lung. The two test groups were statistically identical in terms of age, and exposure period, and overall both groups had very similar mean fiber concentrations and mean fiber sizes. When individual sampling sites within the lung were considered, neither group showed preferential fiber concentration in any area. However, there were definite differences in the intrapulmonary fiber size distribution both within and between the two groups: Cases of mesothelioma showed accumulation of lung fibers in the peripheral upper lobe with shorter central upper lobe fibers. The lung cancer cases demonstrated a reverse pattern, with shorter fibers in the peripheral compared to central upper lobe, but accumulations of long fibers in the peripheral lower lobe. Fiber surfaces and masses showed similar differences among sample sites. We conclude that (1) there is no evidence for fiber concentration variations in different portions of the lung; (2) there is strong evidence for variations in fiber sizes in different portions of the lung, and these differences are most clearly related to fiber length, surface area, and mass; (3) contrary to data from experimental animals, there are no clear gravitational effects on fiber distribution in humans; and (4) there are reproducible differences in intrapulmonary fiber size distribution between mesothelioma and lung cancer cases. These differences may be a manifestation of individual handling of mineral particles because of structural variations in individual lungs. PMID- 2553394 TI - Isolation of similar rolipram-inhibitable cyclic-AMP-specific phosphodiesterases from rat brain and heart. AB - A cyclic AMP phosphodiesterase form of rat brain cytosol was purified by means of affinity chromatography on an immobilized analog of the specific inhibitor rolipram, followed by an exclusion chromatography step. The resulting preparation presented two protein bands in polyacrylamide gel electrophoresis, both with phosphodiesterase activity. Kinetics of cyclic AMP hydrolysis by the purified enzyme proved of the Michaelis type, with a Km of 3 microM, while hydrolysis of cyclic GMP displayed anomalous negatively cooperative kinetics. At micromolar concentrations, this enzyme from hydrolyzed highly specifically cyclic AMP (50 fold faster than cyclic GMP). Cyclic GMP proved a poor competitor of cyclic AMP hydrolysis (Ki 1.04 mM). The neurotropic compound, rolipram, strongly inhibited the enzyme, in a competitive manner (Ki 0.9 microM). This enzyme displayed a molecular mass of around 44 kDa as determined by exclusion chromatography, but two molecular masses of 42 kDa and 89 kDa were observable by electrophoresis on a polyacrylamide gradient gel, compatible with an equilibrium between dimeric and monomeric forms. Isoelectric focusing of the preparation gave rise to two activity peaks of pI 4.8 and 6.7, with identical properties, probably representing two charge isomers of the same protein. An enzyme prepared from rat heart cytosol by the same techniques as for brain phosphodiesterase isolation shared numerous characteristics with the enzyme of cerebral origin, suggesting identity of the rolipram-sensitive form between the two tissues. Since the rolipram-sensitive form detected in crude brain preparations markedly differs from the above-described isolated enzyme, both by its molecular mass in exclusion chromatography and by its pI, it is suggested that an alteration of the native protein, due to dissociation of putative subunits, occurs during the purification procedure. PMID- 2553395 TI - Two types of cytochrome cd1 in the aerobic photosynthetic bacterium, Erythrobacter sp. OCh 114. AB - Components I and II of cytochrome cd1 which had different spectral features were purified from the aerobic photosynthetic bacterium, Erythrobacter sp. strain OCh 114. Component I showed an absorption maxima at 700 and 406 nm in the oxidized form, and at 621, 552.5, 548 and 416 nm in the reduced form. Component II showed an absorption maxima at 635 and 410 nm in the oxidized form and at 628, 552.5, 548 and 417 nm in the reduced form. The relative molecular mass, Mr, of both cytochromes was determined to be 135,000 with two identical subunits. Components I and II showed pI values of 7.6 and 6.8, respectively. The redox potential of hemes ranged from +234 mV to +242 mV, except for the heme d1 of component I (Em7 = +134 mV). Components I and II showed both cytochrome c oxidase and nitrite reductase activities. Cytochrome c oxidase activity was strongly inhibited by a low concentration of nitrite and cyanide. Erythrobacter cytochromes c-551 and c 552 were utilized as electron donors for the cytochrome c oxidase reaction. The high affinity of cytochrome c-552 to component II (Km = 1.27 microM) suggested a physiological significance for this cytochrome. Erythrobacter cytochromes cd1 are unique in their presence in cells grown under aerobic conditions as compared to other bacterial cytochromes cd1 which are formed only under denitrifying conditions. PMID- 2553396 TI - Marker enzymes in rat liver vesicles involved in transcellular transport. AB - In order to label the vesicles involved in transcellular transfer (transcytosis) through hepatocytes, polymeric IgA (pIgA) was conjugated to horseradish peroxidase (HRP) and injected into rats. The endosomes containing this ligand at 10 or 20 min after injection were isolated by the diaminobenzidine-induced density-shift procedure and their content in various marker enzymes was measured. The endosomes carrying pIgA-HRP 10 min after injection contained only traces of 5'-nucleotidase and low amounts of alkaline phosphodiesterase I. The estimated marker enzyme content is similar to that observed for the particles containing galactosylated bovine serum albumin conjugated to HRP, a ligand degraded in lysosomes. However, 20 min after injection, the transcytotic endosomes showed a marked enrichment in 5'-nucleotidase and especially in alkaline phosphodiesterase I. The results confirm the heterogeneity of rat liver endosomes and substantiate the concept of distinct endosomal compartments. PMID- 2553397 TI - Site-directed mutagenesis of the spinach acyl carrier protein-I prosthetic group attachment site. AB - Site-directed mutagenesis was used to change the phosphopantetheine attachment site (Ser38) of spinach acyl carrier protein I (ACP-I) from a serine to a threonine or cysteine residue. 1. Although the native ACP-I is fully phosphopantethenylated when expressed in Escherichia coli, the TH-ACP-I and CY ACP-I mutants were found to be completely devoid of the phosphopantetheine group. Therefore, the E. coli holoACP synthase requires serine for in vivo phosphopantetheine addition to spinach ACP-I. 2. Spinach holoACP synthase was completely inactive in vitro with either the TH-ACP-I or CY-ACP-I mutants. In addition, TH-ACP-I and CY-ACP-I were strong inhibitors of spinach holoACP synthase. 3. The mutant ACPs were weak or ineffective as inhibitors of spinach fatty acid synthesis and spinach oleoyl-ACP hydrolase. 4. Compared to holoACP-I, the mutant apoACP-I analogs had: (a) altered mobility in SDS and native gel electrophoresis, (b) altered binding to anti-(spinach ACP-I) antibodies and (c) altered isoelectric points. The combined physical, immunological and enzyme inhibition data indicate that attachment of the phosphopantheine prosthetic group alters ACP conformation. PMID- 2553398 TI - The free radical of pyruvate formate-lyase. Characterization by EPR spectroscopy and involvement in catalysis as studied with the substrate-analogue hypophosphite. AB - The first-derivative EPR spectrum of the active form of Escherichia coli pyruvate formate-lyase shows an asymmetric doublet with partially resolved hyperfine splittings (g = 2.0037). Isotope substitution studies demonstrated couplings of a carbon-centered unpaired electron to a solvent-exchangeable proton (a = 1.5 mT) and to further hydrogen nuclei (a = 0.36 and 0.57 mT). By selective incorporation of unlabelled tyrosine into 2H-labelled enzyme protein, a tyrosyl radical structure has been ruled out. Circumstantial evidence indicates that the organic free radical, which also displays an ultraviolet absorption signal at 365 nm, is located on a standard amino acid residue of the polypeptide chain. EPR signal quantification found a stoichiometry of 1 spin per active site. The formate analogue hypophosphite has been characterized as a specific kcat inhibitor of pyruvate formate-lyase which destroys the enzyme radical. Protein-linked 1 hydroxyethylphosphonate was previously described as the dead-end product after reaction of the analogue with the intermediary acetyl-enzyme form of the catalytic cycle [W. Plaga et al. (1988) Eur. J. Biochem. 178, 445-450]. EPR spectroscopy of this system has now identified the corresponding alpha-phosphoryl radical as a reaction intermediate [g = 2.0032; a(P) = 2.72 mT, a(3H) = 1.96 mT]; it showed a half-life of about 20 min at 0 degrees C. This finding proves that the enzyme radical is a hydrogen-atom-transferring coenzymic element. PMID- 2553399 TI - Improved purification and biochemical properties of phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis. AB - Monophosphatidylinositol inositol phosphohydrolase (phosphatidylinositol-specific phospholipase C. PtdIns-PLC. EC 3.1.4.10) has been purified from a Bacillus thuringiensis culture supernatant and from the cellular fraction of a recombinant Escherichia coli clone containing the PtdIns-PLC gene from B. thuringiensis. The two-step purification procedure involved ion-exchange chromatography on DEAE Sepharose followed by separation on a Mono-Q/FPLC-column with yields of 32% and 50%, respectively. The molecular mass was determined to be 34 kDa by SDS/PAGE. The isoelectric point of the enzyme was 5.15. The amino-terminal sequences were shown to be identical for the enzymes purified from both organisms. PtdIns-PLC was inhibited by divalent cations using mixed micelles of Triton X-100 and pure phosphatidylinositol. PtdIns-PLC activity was detectable on polyacrylamide gels by activity staining on phosphatidylinostiol-containing agarose. PMID- 2553400 TI - The role of subunit 4, a nuclear-encoded protein of the F0 sector of yeast mitochondrial ATP synthase, in the assembly of the whole complex. AB - The yeast nuclear gene ATP4, encoding the ATP synthase subunit 4, was disrupted by insertion into the middle of it the selective marker URA3. Transformation of the Saccharomyces cerevisiae strain D273-10B/A/U produced a mutant unable to grow on glycerol medium. The ATP4 gene is unique since subunit 4 was not present in mutant mitochondria; the hypothetical truncated subunit 4 was never detected. ATPase was rendered oligomycin-insensitive and the F1 sector of this mutant appeared loosely bound to the membrane. Analysis of mitochondrially translated hydrophobic subunits of F0 revealed that subunits 8 and 9 were present, unlike subunit 6. This indicated a structural relationship between subunits 4 and 6 during biogenesis of F0. It therefore appears that subunit 4 (also called subunit b in beef heart and Escherichia coli ATP synthases) plays at least a structural role in the assembly of the whole complex. Disruption of the ATP4 gene also had a dramatic effect on the assembly of other mitochondrial complexes. Thus, the cytochrome oxidase activity of the mutant strain was about five times lower than that of the wild type. In addition, a high percentage of spontaneous rho- mutants was detected. PMID- 2553401 TI - Characterization and N-terminal amino acid sequence of multiple ferredoxins in kidney and adrenal mitochondria. AB - Two separate ferredoxins that differ in molecular mass by about 1.5 kDa were isolated from both pig kidney and bovine adrenal mitochondria. The proteins had different biochemical and immunological properties and appeared to be distinct gene products. The smaller ferredoxin from pig kidney (renodoxin, Mr approximately equal to 13,500) was very similar to bovine adrenodoxin (Mr = 14,048). Both proteins had nearly identical N-terminal amino acid sequences and electron-transfer activities. However, renodoxin and adrenodoxin expressed distinct antigenic determinants, although they were immunologically cross reactive. The larger kidney (approximately equal to 15-kDa) and adrenal (approximately equal to 15.3-kDa) ferredoxins were biochemically similar to each other but they had lower specific activities and their N-terminal sequences were different when compared to renodoxin and adrenodoxin. Each of the four ferredoxins had a visible absorption spectrum characteristic of a [2Fe-2S] chromophore. But in addition, the larger ferredoxins displayed a prominent A276 peak due to their higher tyrosine content and the presence of tryptophan, which is absent in adrenodoxin and renodoxin. Consequently, the larger ferredoxins were termed Trp-ferredoxin. Using antibody to pig kidney Trp-ferredoxin, the larger adrenal and kidney ferredoxins were found to be very similar immunologically; however, the Trp-containing and adrenodoxin-type ferredoxins did not cross-react in immunoblot analysis. Nevertheless, it was shown from competition ELISA and activity-inhibition analysis that the two ferredoxin types had limited common antigenic determinants. Trp-ferredoxin was the major iron-sulfur protein in kidney whereas adrenodoxin was the dominant molecular form in adrenal gland. PMID- 2553402 TI - Reconstitution of a light-stimulated adenylate cyclase from retina and Neurospora crassa preparations. Characterization of the heterologous systems using normal and degenerative retinas. AB - Adenylate cyclase catalytic subunits from Neurospora crassa membranes may interact with regulatory factors from membranes of bovine retinal rod outer segments (pretreated with N-ethylmaleimide), reconstituting a heterologous system which, in the presence of light, is catalytically active in assay mixtures containing MgATP. Maximal activation was observed at 550 nm. Transducin-depleted retinal membranes were not capable of reconstituting the heterologous light stimulated adenylate cyclase system. Addition of a transducin preparation to depleted membranes restored the reconstitution capacity of these membranes. A similar heterologous adenylate cyclase system was reconstituted with Neurospora and mouse retinal whole membranes (pretreated with N-ethylmaleimide). Membranes from mice suffering photoreceptor degeneration (rd homozygotes) did not reconstitute an heterologous adenylate cyclase system. PMID- 2553403 TI - Synthesis and characterization of mu-conotoxin IIIa. AB - mu-Conotoxin IIIa, a voltage-dependent sodium channel neurotoxin, has been synthesised using solid-phase peptide synthesis employing 9 fluorenylmethoxycarbonyl chemistry. After cleavage from the resin, the peptide was isolated by reverse-phase HPLC and then the six acetamidomethyl groups were removed by treatment with mercuric acetate. The reduced product so formed was purified by reverse-phase HPLC. Protocols were developed to optimize the oxidation of the cysteine residues to form disulphide bonds. Protocols employed using air oxidation together with 2-mercaptoethanol were the most effective. As complete oxidation was never obtained the oxidised peptide was purified by reverse-phase HPLC. The activity of our products was monitored using [3H]saxitoxin binding to eel membranes. The oxidised product was able to completely block [3H]saxitoxin binding in a competitive manner. Lineweaver-Burke analysis of [3H]saxitoxin binding gave a Ki of 1.5 nM, IC50 was determined as 26.6 nM. It was also shown that the pure synthetic mu-conotoxin IIIa had the same retention time on reversephase HPLC as the natural conotoxin IIIa. Thus an active toxin has been synthesised that can be used to probe sodium channels. PMID- 2553404 TI - Mobility of secondary structure units of horse-muscle acylphosphatase. Relation to antigenicity. AB - The antigenic properties of acylphosphatase are compared with its various sequential characteristics (hydrophobicity, chemical shift of the main-chain 1H NMR resonances, numbers and intensities of the nuclear Overhauser enhancements, hydrogen-deuterium exchange and sequential arrangement of the secondary structure units). The discussion is based on the complete sequential assignment of the 1H NMR spectrum and the knowledge of the three-dimensional fold of the protein obtained by NMR spectroscopy from distance geometry calculations. Regions with very different degrees of mobility can be distinguished. It is found that all major antigenic sites are located in the most mobile surface loops. PMID- 2553406 TI - Human cardiac beta-adrenoceptor subtypes: function, regulation and clinical significance. Based on a symposium of the 10th congress of the European Society of Cardiology. August 28-September 1, 1988, Vienna, Austria. PMID- 2553405 TI - Ultrasound compared with computed tomography and pancreatic arteriography in the detection of endocrine tumours of the pancreas. AB - We have evaluated ultrasound, computed tomography and arteriographic findings in 15 patients with 17 endocrine pancreatic tumours having a mean diameter of 2.3 cm (range 1-7 cm). All patients under-went computed tomography, and all but one ultrasound and arteriography. Ultrasound was the initial investigation in 11 patients, and identified 10 of the 16 tumours present in 14 patients. Two tumours were found at ultrasound reexamination after having been identified by other radiological methods. Computed tomography revealed 8 out of 17 tumours, while arteriography identified 8 out of 16 tumours. Computed tomography was the initial investigation in 4 patients, and identified one tumour. In only 4 patients were tumours not detected by any of the imaging methods. The sensitivities of ultrasound, computed tomography and arteriography in the detection of pancreatic tumours were 62.5% (95% confidence interval 50.4-74.6%), 47.1% (95% confidence interval 35.0-59.2%), and 50.0% (95% confidence interval 37.5-62.5%), respectively. Ultrasound was thus more accurate than computed tomography or arteriography in detecting endocrine pancreatic tumours, and should be the initial radiological investigation. PMID- 2553407 TI - Coexistence and localization of beta 1- and beta 2-adrenoceptors in the human heart. AB - Quantitative autoradiography was used to determine the location and density of beta 1- and beta 2-adrenoceptors in the right atrium (RA), left ventricular free wall (LVFW), right ventricular free wall (RVFW), interventricular septum (IVS), right atrium from an area near the atrioventricular node (RAAV) and cardiac nerves (N) taken from a patient with end-stage cardiac failure. The densities of beta-adrenoceptors detected by the non-selective beta-adrenoceptor antagonist radioligand (-)-[125I] cyanopindolol (50pM) were 4.93 (N), 10.6 (RVFW), 12.2 (RA), 12.4 (IVS), 15.8 (LVFW) and 18.7 fmol (mg protein)-1 (RAAV). The proportion of beta 2-adrenoceptors ranged from 19.5% (RAAV) to 95% (N). RA taken from patients with ischaemic heart disease had a higher density of beta-adrenoceptors (29.3 fmol (mg protein)-1). The results suggest that both beta 1- and beta 2 adrenoceptors are down-regulated in patients with end-stage cardiac failure. Positive inotropic responses were established to (-)-isoprenaline, RO363 (beta 1 selective), procaterol (beta 2-selective) and dopexamine in the absence or presence of the antagonist CGP 20712A (beta 1-selective) or ICI 118,551 (beta 2 selective) in electrically driven human right atrial appendage strips. RO363 and procaterol were nearly full agonists in this preparation and produced their responses through activation of beta 1- or beta 2-adrenoceptors, while dopexamine was a partial agonist which produced its inotropic responses through activation of both receptor subtypes. These studies demonstrate the presence and location of beta 1- and beta 2-adrenoceptors in the human heart. PMID- 2553408 TI - Beta-adrenoceptor regulation in the human heart: can it be monitored in circulating lymphocytes? AB - In heart failure a decrease in cardiac beta-adrenoceptors presumably due to endogenous down-regulation by the elevated catecholamines is a general phenomenon. Thus, attempts have been made to assess beta-adrenoceptor function in patients with chronic heart failure in order to monitor the functional state of cardiac beta-adrenoceptors. The model most widely used is that of circulating lymphocytes that contain a homogeneous population of beta 2-adrenoceptors coupled to the adenylate cyclase/cyclic AMP system. The biochemical and pharmacological properties of beta 2-adrenoceptors present in lymphocytes are quite comparable to those of beta 2-adrenoceptors in the human heart, but clearly different from those of human cardiac beta 1-adrenoceptors. Furthermore, beta-adrenoceptor agonists and antagonists regulate lymphocyte beta 2- and cardiac beta 1- and beta 2-adrenoceptors in a subtype-selective fashion: while non-selective agonists (independent of exogenously applied or endogenously elevated) and antagonists affect both cardiac beta 1- and beta 2- as well as lymphocyte beta 2 adrenoceptors, beta 1-selective agonists and antagonists influence only cardiac beta 1-, but not cardiac and lymphocyte beta 2-adrenoceptors. Finally, direct comparison of lymphocyte and cardiac beta-adrenoceptor densities revealed that changes in lymphocyte beta 2-adrenoceptors are significantly correlated with changes in cardiac beta 2-adrenoceptors, but not related to changes in cardiac beta 1-adrenoceptors. Since beta 1-adrenoceptors predominate in all parts of the human heart, the use of lymphocyte beta 2-adrenoceptors as a tool for predicting the status of cardiac beta-adrenoceptors is, therefore, quite limited. PMID- 2553409 TI - In vivo studies on the function of cardiac beta-adrenoceptors in man. AB - The original Lands classification considered cardiac beta-adrenoceptors to be predominantly beta 1 in type and to respond to both noradrenaline and adrenaline. Radioligand binding studies subsequently identified substantial numbers of beta 2 adrenoceptors in cardiac tissue. Studies in man employing intensive exercise, isoprenaline testing and a variety of selective and non-selective beta adrenoceptor antagonists would now suggest that under certain circumstances these receptors may be functionally active. Severe exercise, as a sympathetic stimulus, is associated with high circulating noradrenaline concentrations, and appears to produce tachycardia predominantly by cardiac beta 1-adrenoceptor activity. In contrast, isoprenaline testing increases heart rate by a number of mechanisms: it stimulates beta 1-adrenoceptor activity in the sinoatrial node; it may result in reflex vagal withdrawal; and it appears to stimulate cardiac beta 2-adrenoceptors directly. In addition, isoprenaline also facilitates noradrenaline release by presynaptic beta 2-adrenoceptors. Thus the effects of isoprenaline would appear to mimic stress in man with the subsequent release of adrenaline from the adrenal medulla and its complex interaction directly and indirectly with both cardiac beta 1 and beta 2-adrenoceptors. These differing circumstances may also influence the relative efficacy of beta 1-selective and non-selective beta-adrenoceptor agonists and antagonists. PMID- 2553410 TI - Drug- and disease-induced changes of human cardiac beta 1- and beta 2 adrenoceptors. AB - Cardiac beta-adrenoceptor density and subtype distribution has been determined in different kinds of heart failure. A decrease in cardiac beta-adrenoceptor function appears to be a general phenomenon in all kinds of heart failure. However, cardiac beta 1- and beta 2-adrenoceptors seem to be differentially affected in different kinds of heart failure: while in end-stage idiopathic dilated cardiomyopathy the diminished cardiac beta-adrenoceptor function is due to a selective loss in beta 1-adrenoceptors, in mitral valve disease, tetralogy of Fallot and end-stage ischaemic cardiomyopathy it is characterized by a concomitant reduction in beta 1- and beta 2-adrenoceptors. Chronic treatment of heart failure patients with beta-adrenoceptor antagonists leads to an up regulation of cardiac beta-adrenoceptors, but in a subtype-selective fashion: beta 1-selective antagonists increase only cardiac beta 1-adrenoceptors, whereas non-selective antagonists increase both beta 1- and beta 2-adrenoceptors. Such a (subtype-selective) 'recovery' of cardiac beta-adrenoceptors may be one reason for the beneficial effects of low-dose beta-adrenoceptor antagonist treatment in patients with severe heart failure. PMID- 2553412 TI - Cardiodepressive effects of antiarrhythmic drugs. AB - Negative inotropic effects of antiarrhythmic drugs are ultimately mediated by alterations of intracellular Ca++. With class I antiarrhythmic drugs a strong correlation between the Na+ inward current and the Ca++ influx exists whereby Ca++ influx is changed by an exponent, n, of approximately 3. Generally, the longer the duration of Na+ -channel blockade the more pronounced is negative inotropy. Antiarrhythmic drugs of class I may be subdivided into those with a short recovery time of Na+ channels (lidocaine-type), those with an intermediate recovery time, such as quinidine and newer antiarrhythmics such as flecainide and propafenone, and those with a recovery time greatly exceeding 1 s, such as disopyramide and prajmaline. The degree of cardiodepression is modulated by peripheral circulatory effects such as vasoconstriction and vasodilation. In order to differentiate between general cardiodepressive effects and true negative inotropy, sophisticated methods, for example, volume-pressure relationship curves, have to be applied to determine the degree of pre- and afterload alterations, also accounting for cardiodepressive effects. The net effect of cardiodepression is due to several factors: true negative inotropy, additional influences on the peripheral circulation, the functional state of the myocardium, and the beneficial or detrimental effects on the prevailing arrhythmia. Based on these theoretical considerations, combined drug therapy of arrhythmias has to be installed under the aspect of cardiodepressive side-effects, especially in an already impaired heart. PMID- 2553411 TI - Pharmacology of positive inotropic phosphodiesterase III inhibitors. AB - Cardiac phosphodiesterase III (PDE) inhibitors derived from pyridinone, imidazolone, pyridazinone and related structures form a new class of positive inotropic vasodilator agents (e.g. milrinone) that are beneficial in the treatment of acute and chronic heart failure. These agents inhibit the intracellular hydrolysis of cyclic AMP, thereby promoting cyclic AMP-catalysed phosphorylation of sarcolemmal calcium channels and activating the calcium pump. Drugs such as milrinone have a wider therapeutic index than the cardiac glycosides. They also have vasodilator and lusitropic actions and are devoid of the central stimulant actions that narrow the therapeutic index of theophylline and other methylxanthines. Receptor down-regulation, which curtails the inotropic efficacy of beta-adrenoceptor agonists, does not compromise the efficacy of PDE inhibitors. The effectiveness of these new agents is, however, dependent upon some degree of basal adenylate cyclase activity. Individual PDE inhibitors differ in terms of both chronotropic and extracardiac properties. The reasons for this are not yet fully understood. PMID- 2553413 TI - Ovarian malignant mixed mesodermal tumor response to cis diamminedichloroplatinum. AB - A patient with primary, bulky, Stage III, malignant mixed mesodermal tumor (MMMT) of the ovary achieved a partial response to postoperative cis Diamminedichloroplatinum (DDP). These rare tumors have previously been relatively resistant to chemotherapeutic agents. The use of DDP resulted in significant palliation in our patient and should be strongly considered in the postoperative therapy of patients with MMMT of the ovary. PMID- 2553414 TI - Clinico-pathological profile of endometrial carcinoma in young women (under 40 years of age). AB - Twenty primary endometrial carcinomas in 40 year-old or younger women were collected from a series of 845 carcinomas and newly re-evaluated. The typical histological profile was well-differentiated adenocarcinoma (30%) or adenoacanthoma (35%); however, a case of adenosquamous carcinoma and an exceptional malignant mixed mesodermal tumor were also observed. No tendency for myometrial infiltration (M0) was displayed by 11 (55%) of the cases, while 6 were M1 and 3 were M2 infiltrating neoplasias. Adenomatous hyperplasia was associated with cancer in 53% of the 15 assessable cases. Histopathological diagnostic problems were elicited in 5 cases only. Typical Stein-Leventhal syndrome occurred in 7 patients (35%), while polycystic ovaries were observed in 15 cases (75%). Usually, the patients were obese (40%) and nulligravida (65%). All but one case were stage I disease. Prognosis seems to be excellent following surgery. PMID- 2553415 TI - "False positive" immunoscintigraphic diagnosis of liver abscesses in a patient with a necrotic liver tumour. AB - The following report describes the scintigraphic findings in a patient who underwent immunoscintigraphy with anti granulocyte Mab because of septic fever. Focal tracer accumulation in the liver corresponding with hypoechoic and hypodense areas documented by sonography and computed tomography was believed to represent liver abscesses, yet surgery and autopsy revealed a large necrotic cholangiocellular liver carcinoma. The possible mechanisms, which had led to the false positive immunoscintigraphic image are discussed and we conclude that a necrotic neoplasm has to be a major differential diagnosis in any case of positive liver uptake in an anti granulocyte scan. PMID- 2553416 TI - Scintigraphic rib abnormality mimicking metastasis in a breast cancer patient. AB - A breast cancer patient revealed an abnormal bone scan in the ribs caused by an unusual form of indirect trauma (bras). The recognition of the phenomenon was important in order to avoid the misinterpretation of the scintigram and consequently, to precede an inadequate therapeutic decision. PMID- 2553418 TI - Analysis of hemodynamics and blood gas in relation to blood ketone body ratio in partially hepatectomized patients. AB - A total of 205 studies of hemodynamics and blood gas analyses were performed in relation to hepatic mitochondrial redox state in 23 partially hepatectomized patients. When the blood ketone body ratio (the ratio of acetoacetate to beta hydroxybutyrate in arterial blood) decreased, that is when the hepatic mitochondrial redox state was reduced, a hyperdynamic state with increased cardiac index, decreased systemic and pulmonary vascular resistance together with decreased oxygen consumption was observed in spite of the increased oxygen availability. Inhibited oxidative metabolism in the Krebs cycle may have resulted in the decreased oxygen consumption and decreased vascular tone. This study supports the concept that the hepatic metabolic derangements due to reduced hepatic mitochondrial redox state may be a factor in the decreased oxygen consumption and hyperdynamic state. PMID- 2553417 TI - Benzodiazepine receptors increase in post-mortem brain of chronic schizophrenics. AB - [3H]-Flunitrazepam (FNT) binding was measured in the post-mortem brains of 13 chronic schizophrenics and 10 controls whose mean ages and death-to-freezing intervals were the same in each group. The specific binding of [3H]-FNT to the medial frontal cortex, orbitofrontal cortex, orbital cortex, medial and inferior temporal gyri, superior temporal gyrus, cornu Ammonis 1-3 and putamen was significantly higher in schizophrenics than in controls. Specific binding to the eye movement area (frontal eye field), motor cortex, lateral occipitotemporal gyrus, dentate gyrus of the hippocampus and secondary and tertiary visual cortex did not differ in the two groups. Type 1 benzodiazepine (BZ) binding sites in the superior temporal gyrus of schizophrenics, determined from the displacement of [3H]-FNT binding using a triazolopyridazine, CL 218,872 (200 nM), were significantly higher than in the control group. The increase in type 2 BZ binding sites was not significant. Antipsychotic or benzodiazepine medication did not appear to affect the results. There were significant correlations between specific [3H]-FNT binding and concentration of GABA (positive) and of glutamic acid (negative), specific [3H]-kainic acid binding (positive), activity of tyrosine hydroxylase (positive), and substance P-like immunoreactivity (positive) in many areas of the brain. The Bmax of [3H]-spiperone binding in the putamen was also correlated positively with specific [3H]-FNT binding. These data suggest that dysfunction of BZ receptors may be involved in the pathogenesis and some symptoms of chronic schizophrenia. PMID- 2553419 TI - Phase II study of pirarubicin in advanced non-small cell lung cancer. PMID- 2553420 TI - Phase II trial of buserelin in hepatocellular carcinoma. PMID- 2553421 TI - Immunoprophylaxis of cytomegalovirus infections in transplanted patients. AB - Cytomegalovirus causes severe infections in immunosuppressed patients and anti viral treatments remain unsatisfactory. In an attempt to prevent life-threatening CMV infections, immunoprophylaxis using hyperimmune immunoglobulins has been studied but led to conflicting results in the literature. The role of passive immunization in allogeneic marrow and renal transplant patients is discussed. PMID- 2553422 TI - Lipolysis of polyenoic fatty acid esters of human chylomicrons by lipoprotein lipase. AB - Human chylomicrons, obtained from chylous pleural fluid of a patient with mesothelioma, were incubated with bovine milk lipoprotein lipase. No difference in the release of different C16-C18 fatty acids as unesterified fatty acids, or in their appearance in diacylglycerols, was observed. The rate of lipolysis of arachidonic (20:4), and eicosapentaenoic (20:5) acid ester bonds of chylomicron triacylglycerols was slower, and a larger proportion of these fatty acids accumulated in mono- and diacylglycerols. The hydrolysis pattern for docosahexaenoic acid (22:6) was intermediary between that of the C16-C18 fatty acids and that of 20:4 and 20:5. The data suggest that the presence of a double bond at position 5 in the eicosanoid precursors may be important for the rate of lipolysis. The relative resistance of the 20:4- and 20:5- esters of chylomicrons to lipoprotein lipase may be important for the tissue distribution of these fatty acids. PMID- 2553423 TI - Effects of a bolus dose of atrial natriuretic factor in young and elderly volunteers. AB - We assessed the haemodynamic and renal effects as well as the effects on plasma cGMP levels of a small i.v. dose (33 micrograms) of human atrial natriuretic factor (99-126; hANF) in two age groups of healthy volunteers. Binding properties of platelet ANF receptors were also measured. The elderly (four males, eight females, mean age 52.3 years) showed increased haemodynamic (decrease in blood pressure) and renal responses (diuresis, natriuresis, calciuresis) as well as greater increases in plasma cGMP levels and urinary cGMP excretion than the young subjects (four males, 12 females, mean age 26 years). Binding capacities and affinities of platelet ANF receptors were identical in both groups. These data indicate that the sensitivity to ANF increases with age and that this increased sensitivity is reflected in the reactivity of plasma cGMP levels but not in the properties of platelet ANF receptors. The data may be important for the therapeutic use of ANF, for the understanding of the physiological regulation of ANF action and may underline the necessity of using age-matched control subjects for clinical studies on the possible therapeutic effectiveness of ANF. PMID- 2553424 TI - Intravenous atrial natriuretic peptide does not affect water and ion transport in the human small intestine. AB - Atrial natriuretic peptide (ANP) increases renal sodium and water excretion in several species including man. In rats ANP was also found to influence water and electrolyte transport in the small intestine. In the present study we investigated whether ANP can alter transport in the jejunum and ileum of healthy volunteers using a triple-lumen perfusion technique. The small intestine was perfused under steady-state conditions with a plasma-like electrolyte solution using polyethylene glycol as a nonabsorbable volume marker. After an initial control period with intravenous saline infusion ANP was administered intravenously at a dose of 400 pmol kg-1 h-1. This dose led to a significant (P less than 0.05) increase in the plasma levels of ANP (up to 22-fold) and cGMP (up to 15-fold), and of urine volume. Intestinal water and electrolyte transport were, however, not affected by ANP. Our results suggest that circulating ANP does not play a role in the regulation of mucosal water and ion transport in the human small intestine. PMID- 2553426 TI - Is the sumatriptan (GR 43175)-induced endothelium-dependent relaxation of pig coronary arteries mediated by 5-HT1D receptors? PMID- 2553425 TI - Renal responsiveness to synthetic human parathyroid hormone 1-38 in healthy subjects. AB - Renal responsiveness to synthetic human parathyroid hormone (HPTH) 1-38 was examined in seven healthy volunteers. Each subject received three different doses (35 micrograms, 17.5 micrograms, 8.75 micrograms) as bolus intravenous injections. Rapid, transient, dose-dependent decreases in diastolic blood pressure and increases in heart rate were observed, as reported previously in animal studies, suggesting an acute vasodilatory effect of the hormone. The extracellular cyclic AMP responses were brisk and dose-dependent in the group as a whole as well as in the individual subjects. In contrast, the phosphaturic responses, though statistically significant with all three doses, were of a lesser magnitude and showed an individual variability. Nearly maximal decreases in TmP/GFR were found even after the lowest dose with minimal increases in cyclic AMP in some subjects. The injections of HPTH 1-38 also induced a dose-dependent natriuresis which was accompanied by an increase in excreted calcium. It seems that under these experimental conditions the sodium-dependent renal calcium transport cannot be compensated by the effects of the hormone on the distal tubule. HPTH 1-38 can be safely and predictably used for the assessment of renal responsiveness to PTH in routine clinical practice. PMID- 2553427 TI - The C-terminal hexapeptide, endothelin-(16-21), discriminates between different endothelin receptors. PMID- 2553429 TI - Effects of Hoe 065, a compound structurally related to inhibitors of angiotensin converting enzyme, on acetylcholine metabolism in rat brain. AB - Hoe 065, a compound structurally related to inhibitors of angiotensin converting enzyme, caused a fall in the content of acetylcholine (ACh) in different brain areas of the rat following i.p. administration in the range 0.03-30 mg/kg. This effect occurred 0.5 h after a single injection and lasted for at least 6 h. Simultaneous administration of the choline uptake inhibitor hemicholinium-3 (HC 3) with Hoe 065 potentiated the decrease in ACh content induced by HC-3. In the same dose range Hoe 065 acutely enhanced the activity of the enzyme choline acetyltransferase as well as the capacity of the high-affinity choline uptake system which is considered as the rate-limiting step in the synthesis of ACh. Cholinesterase activity in vivo was not altered by the compound. Hoe 065 produced a concurrent elevation of brain cyclic GMP content. Taken together, these results suggest that Hoe 065 acutely increases cholinergic activity within its physiological range, probably by means of an enhanced release of ACh. PMID- 2553428 TI - Diethyldithiocarbamate and disulfiram inhibit MPP+ and dopamine uptake by striatal synaptosomes. AB - Diethyldithiocarbamate (DDC) was found to inhibit the uptake of both dopamine and 1-methyl-4-phenyl-pyridinium ion (MPP+, the putative toxic metabolite of the neurotoxicant MPTP) by striatal synaptosomes. Disulfiram, the corresponding disulfide of DDC, was effective at concentrations 1,000 times lower (10(-6) vs. 10(-3) M). Disulfiram, but not DDC, reacted very efficiently with synaptosomal protein thiols; both DDC- and disulfiram-induced uptake inhibition could be reversed by the thiol-reducing agent dithiothreitol. Two other thiol-reactive compounds, N-ethylmaleimide (NEM) and p-hydroxymercuribenzoate (PMB), also impaired the uptake of MPP+ by striatal synaptosomes. PMB, which does not cross membranes, was even more potent than the lipophilic NEM in blocking MPP+ uptake. These results suggest that (1) the effect of DDC may be mediated by disulfiram, (2) the uptake of MPP+ and dopamine by striatal synaptosomes is dependent on the redox state of protein thiols, and (3) these protein thiols are located at the outer surface of synaptosomal membranes. PMID- 2553430 TI - Biphasic effect of a kappa-opioid receptor agonist on plasma oxytocin levels in rats. AB - The effect of the kappa-opioid receptor agonist, bremazocine, on plasma oxytocin levels in rats was measured by a sensitive radioimmunoassay. Initially, a decrease in plasma oxytocin levels was seen 30 min after injection. This was in accordance with the bremazocine inhibition of oxytocin release after submaximal electrical stimulation seen in isolated neurointermediate lobes. The initial decrease in plasma oxytocin reversed, and 4 h after injection of bremazocine a 20 fold increase in the oxytocin level was seen. The rise in plasma oxytocin was paralleled by a rise in plasma sodium. The biphasic time course of the plasma oxytocin response can be explained by a combination of an inhibition of oxytocin release from the neurohypophysis and an increased water excretion leading to an elevation in plasma sodium, which may be responsible for the late rise in plasma oxytocin. Down-regulation of the opioid receptors may also contribute to the delayed rise in plasma oxytocin. PMID- 2553431 TI - Growth hormone and insulin-like growth factor I stimulate Leydig cell steroidogenesis. AB - Leydig cells from 40 days old rats were incubated with or without human growth hormone (hGH) or insulin-like growth factor I (IGF-I) in the presence or absence of human chorionic gonadotropin (hCG), and testosterone and cyclic AMP (cAMP) levels in the medium were measured. Neither hGH nor IGF-I increased testosterone production in the absence of hCG in concentrations up to 1000 and 100 ng/ml, respectively. However, both peptides increased hCG-induced testosterone production in a dose-dependent manner. The maximal stimulatory concentrations of hGH and IGF-I were 100 and 50 ng/ml, respectively. Human GH did not further enhance the IGF-I-stimulated steroidogenesis. The hGH-augmented steroidogenesis was inhibited by anti-hGH IgG and anti-IGF-I IgG. hGH also enhanced hCG stimulated cAMP production time dependently, suggesting that the stimulatory effect of hGH on steroidogenesis was due to an increased cAMP production. These data suggest that the effect of hGH might be mediated by locally produced IGF-I, which may act as a modulator on gonadal development in the presence of gonadotropin. PMID- 2553432 TI - GABAB receptors are up-regulated by chronic treatment with lithium or carbamazepine. GABA hypothesis of affective disorders? AB - The effects of lithium and carbamazepine on GABAA and GABAB receptors were examined. The binding of [3H]muscimol and [3H](-)-baclofen to synaptic membranes from rat brain was used to label GABAA and GABAB receptors, respectively. Neither the [3H]muscimol nor the [3H](-)-baclofen binding site was displaced by lithium or carbamazepine even at a concentration of 100 microM. A single treatment with either of these drugs did not induce any change in [3H]muscimol and [3H](-) baclofen binding sites in the frontal cortex and hippocampus. [3H](-)-Baclofen binding sites were up-regulated in the hippocampus but not in the frontal cortex following chronic treatment with lithium or carbamazepine. These results suggest that one common mechanism of action of lithium and carbamazepine is mediated by GABAB receptors and that GABA is involved in the pathophysiology of affective disorders. PMID- 2553433 TI - Comparison of the behavioral and biochemical effects of the NMDA receptor antagonists, MK-801 and phencyclidine. AB - The behavioral and biochemical effects of the noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist, MK-801 [+)-5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine maleate) were compared with those of phencyclidine (PCP). In the dose range used in this study, MK-801 (0.125-0.5 mg/kg i.p.) produced ataxia and other behavioral responses which were similar to PCP (5-10 mg/kg i.p.). However, turning and backpedalling induced by MK-801 were not dose-dependent and less intense at the dose producing approximately the same level of ataxia as PCP. Neurochemically, MK-801 (0.5 mg/kg i.p.) increased dopamine turnover in the cortex and striatum, but had no effect on 5-HT systems. It was also 3.4 times less potent in inhibiting 5-HT uptake than PCP. These results suggest that the behavioral responses induced by MK-801 involve primarily the PCP receptor and the dopamine system, and that the differences from PCP reflect a reduced effect on the 5-HT neuronal system. PMID- 2553434 TI - Cyclic nucleotides augment the phasic action of tetraethylammonium on guinea-pig trachealis. AB - It is known that after exposure to (3 mM) tetraethylammonium (TEA), guinea-pig isolated trachealis generates tonic spasm followed by phasic tension changes. We found that 8-Br-cAMP or 8-Br-cGMP (10(-6)-5 x 10(-4) M) suppressed tonic spasm secondary to TEA and induced rapid mechanical oscillations. NaN3, forskolin, aminophylline, and isoprenaline had effects similar to those of the cyclic nucleotides. Tetrodotoxin, atropine, diphenhydramine, and cimetidine were without effect on the rapid oscillations. KCl (16 mM) stimulated the oscillations in tissue treated with TEA or TEA plus 8-Br-cGMP. Diltiazem abolished all rhythmic activity in TEA plus 8-Br-cGMP-treated tissues. We conclude: (A) The rapid oscillations are a result of increased cytoplasmic calcium flux with K+ affecting the contractile arm by increasing Ca2+ entry and cyclic nucleotides affecting the relaxant arm by reducing cytosolic free Ca2+. (B) The cyclic nucleotide effect is direct. (C) Cyclic nucleotides may enhance the rate of contraction and relaxation in the presence of some forms of phasic activity. PMID- 2553435 TI - Magnitude of opioid dependence after continuous intrathecal infusion of mu- and delta-selective opioids in the rat. AB - The continuous intrathecal infusion of morphine (2, 6, 20 nmol/h), sufentanil (0.06, 0.2, 0.6 nmol/h), [D-Ala2,MePhe4, Gly-ol5]enkephalin (DAMGO) (0.1, 0.3, 1.0 nmol/h) or [D-Ala2,D-Leu5]enkephalin (DADLE) (2, 6, 20 nmol/h) in unanesthetized rats produces a dose-dependent increase in hot plate latency 1 day after pump implant followed by a gradual return to baseline values by days 3-4, i.e. tolerance. Rats assessed for opioid dependence after 7 days of intrathecal (i.t.) infusion of opioids show a withdrawal syndrome most readily noted by withdrawal body shakes (WBS) after injection of the opioid antagonist, naloxone (1 mg/kg i.p.). The number of WBS was proportional to the infusion dose of opioid agonist. Although each tolerance-producing agent was infused in one of three log spaced (low, medium, high) doses, selected to have approximately equal antinociceptive activity across agents, the agents varied in the apparent degree of dependence. Thus, at the highest infusion dose, the average number of WBS observed was greatest for DADLE (32.8), morphine (30.2) and sufentanil (25.0) while animals treated with DAMGO displayed a significantly less degree of opioid dependence (8.7). PMID- 2553436 TI - Opioid receptor affinities of the blood-derived tetrapeptides hemorphin and cytochrophin. AB - Hemorphin-4 and cytochrophin-4 displayed affinities for mu- and delta-opioid receptors that were in the same range as those observed for the structurally related beta-casomorphins. However, they showed markedly higher affinities at kappa-opioid binding sites when compared to the beta-casomorphins. These blood derived peptides could be involved in blood pressure regulation. PMID- 2553437 TI - First use of a beta-carboline as photoaffinity label for the benzodiazepine receptor. AB - Photolabelling of benzodiazepine receptors isolated from rat cortex with a new beta-carboline-type photoaffinity label, ethyl 6-azido-beta-carboline-3 carboxylase, at 254 nm produced a 42% decrease in the maximal number of propyl beta-carboline-3-carboxylate binding sites but practically no decrease in the number of flunitrazepam binding sites. Moreover, the binding affinity of ethyl beta-carboline-3-carboxylase was diminished 11-fold by photolabelling while that of diazepam was diminished less than 2-fold. These results provide additional evidence that beta-carbolines and benzodiazepines bind to discrete sites on the benzodiazepine receptor. PMID- 2553438 TI - Comparative neuroanatomical distribution of the kappa and mu opioid receptors in guinea pig brain sections. AB - The opiate family of receptors is believed to consist of the mu, delta, kappa and sigma subtypes. Autoradiographic studies have been conducted to define the distribution of the mu and delta receptors. However, due to the lack of a selective ligand definitive studies have not been reported for the kappa receptor. Recently such a ligand was discovered and characterized. Results are presented in which the distribution of kappa receptors in guinea pig brain slices were determined using autoradiographic techniques and its distribution compared to that of a typical mu ligand. The results demonstrate numerous differences between the two opioid systems. The kappa receptors predominate in the prefrontal cortex, the nucleus accumbens, and deep cortex; whereas the mu receptors have a more intense, broader and different distribution. PMID- 2553439 TI - Cross-desensitization of endothelin- and sarafotoxin-induced phosphoinositide turnover in neurons. PMID- 2553440 TI - Trans-ACPD, a selective agonist of the phosphoinositide-coupled excitatory amino acid receptor. PMID- 2553441 TI - Memantine is a potent blocker of N-methyl-D-aspartate (NMDA) receptor channels. PMID- 2553442 TI - Pharmacological activities of optically pure enantiomers of the kappa opioid agonist, U50,488, and its cis diastereomer: evidence for three kappa receptor subtypes. AB - De Costa et al. (FEBS Lett. 223, 335; 1987) recently described the synthesis of optically pure enantiomers of (+/-)-trans-3,4-dichloro-N-methyl-N-[2-(1 pyrrolidinyl)-cyclohexyl] benzeneacetamide (U50,488). In the present study we examined the in vitro opioid receptor selectivity of (-)-(1S,2S)-U50,488, (+) (1R,2R)-U50,488 and (+/-)-cis-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl) cyclohexyl] benzeneacetamide (the cis diasteromers of U50,488), as well as their pharmacological activities in rhesus monkeys. Using [3H]5 alpha,7 alpha,8 beta-( )-N-methyl-N-[7-(1-pyrrolidinyl)-1-oxaspiro (4,5)dec-8-yl]-phenyl benzeneacetamide ([3H]U69,593) to label kappa binding sites of guinea pig membranes, the apparent dissociation constants of the enantiomers of U50,488 were 0.89 and 299 nM, for the (S,S) and (R,R) enantiomers, respectively. The (-)-cis and (+)-cis diastereomers had apparent Kds of 167 and 2715 nM, respectively. Binding surface analysis of the interaction of (-)-(1S,2S)-U50,488 with kappa binding sites labeled by [3H]bremazocine resolved two binding sites at which (-) (1S,2S)-U50,488 had Kds of 30 and 10,485 nM, respectively. The (+/-)-cis, (-)-cis and (+)-cis diastereomers of U50,488 (1 microM) did not inhibit [3H]bremazocine binding. Rhesus monkeys were trained to discriminate ethylketocyclazocine (EKC) and saline. All compounds tested substituted completely for EKC. The order of potency was (-)-(1S,2S)-U50,488 greater than (+/-)-U50,488 greater than (+/-)-cis diastereomer of U50,488 greater than (+)-(1R,2R)-U50,488.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553443 TI - High NaCl diets increase alpha 2-adrenoceptors in renal cortex and medulla of NaCl-sensitive spontaneously hypertensive rats. AB - Diets high in NaCl simultaneously elevate renal alpha 2-adrenoceptor binding and exacerbate hypertension in young NaCl-sensitive spontaneously hypertensive rats (SHR-S). The present study tests the hypothesis that in SHR-S on a high NaCl diet, an upregulation of renal alpha 2-adrenoceptors is present in densely innervated areas of the kidney, and this precedes the increase in blood pressure. Seven week old SHR-S fed on a high (8%) compared to basal (1%) NaCl diet for 2 weeks displayed significantly exacerbated hypertension and elevated renal alpha 2 adrenoceptor binding in both cortex and medulla. In contrast one week on the high NaCl diet did not alter renal alpha 2-adrenoceptor number or blood pressure in SHR-S. Autoradiographic experiments demonstrated that the NaCl-induced upregulation of alpha 2-adrenoceptors occurs in all areas of the renal cortex and medulla. None of these differences were observed in NaCl-resistant, Wistar-Kyoto rats (WKY). Further, the high NaCl diet did not alter renal alpha 1-adrenoceptor binding in SHR-S or WKY. Together with previous findings, these data suggest that the NaCl-induced upregulation of renal alpha 2-adrenoceptors is not specific to densely innervated regions of the kidney. PMID- 2553444 TI - Evidence for a common site of action of lidocaine and carbamazepine in voltage dependent sodium channels. AB - The finding that the development of lidocaine-kindled seizures is blocked by carbamazepine suggests an interaction of carbamazepine with local anesthetic mechanisms. To study the site of interaction, the effects of lidocaine, carbamazepine and another anticonvulsant drug, phenytoin on scorpion venom enhanced specific binding of [3H]batrachotoxinin A 20-alpha-benzoate to the sodium channel gating complex were examined in vitro in a rat brain hippocampus preparation. Lidocaine shifted the concentration inhibition curve of carbamazepine to the right and vice versa. Carbamazepine shifted the concentration inhibition curve of phenytoin to the right and vice versa. The experimentally determined apparent dissociation constants were in a good agreement with the dissociation constants calculated for a one-site model, suggesting that the interaction occurs because lidocaine shares a common binding site with carbamazepine and phenytoin in the voltage-dependent sodium channels. PMID- 2553445 TI - Stimulation by phaclofen of inositol 1,4,5-triphosphate production in cultured neurons from chick tectum. AB - The influence of gamma-aminobutyric acid (GABA) receptor agonists and antagonists on the intracellular concentration of inositol 1,4,5-triphosphate (IP3) was examined in neuronal cultures of embryonic chick tectum. GABA and selective agonists of its receptors were inactive, whereas phaclofen, a GABA-B receptor antagonist, increased IP3 levels in a concentration-dependent manner. This effect occurred at phaclofen concentrations much lower than those required to affect GABA-B receptors. It is concluded that phaclofen, in addition to its known effect at GABA-B receptors, is also active at an as-yet undefined site. PMID- 2553446 TI - A possible mechanism of action of tetramethylpyrazine on vascular smooth muscle in rat aorta. AB - The vasodilatation of isolated rat aorta by tetramethylpyrazine (TMP) was studied by examining its effect on phenylephrine-induced contraction. We found no difference between the effects on intact and on endothelium-denuded preparations. The effect of TMP was similar to that of theophylline because propranolol did not block the vasodilatation. Also, there was a summation effect when the pyrazine was combined with theophylline. Furthermore, like that due to theophylline, the vasodilatation was accompanied by an increase in cyclic AMP. The pyrazine, as do other dilators, affected differently the two separate phases of the contractile response elicited with either phenylephrine or high potassium. The drug predominantly suppressed the phasic responses but both the phasic and tonic phases could be inhibited significantly if the concentration of the pyrazine was high enough. The present results suggest that intracellular accumulation of cyclic AMP and blockade of the release of calcium from internal stores may be important elements of the mechanism by which TMP reduces the development of tension in rat aortic smooth muscle. PMID- 2553447 TI - Functional coupling of GABAA receptors and benzodiazepine recognition site subtypes in the spinal cord of the rat. AB - The interaction between GABAA receptors and benzodiazepine (BZD) recognition site subtypes in the spinal cord of the rat was investigated. Computer analysis of displacement curves for [3H]flunitrazepam [( 3H]FNT) binding by 2-oxo-quazepam (2OXOQ) indicated the presence of two subtypes of BZD recognition sites in this region. Type I sites accounted for approximately 25% of the total number of BZD recognition sites, the remainder being Type II sites. A similar proportion of Type I and Type II sites was obtained by Scatchard analysis of the saturation curves for [3H]FNT, [3H]2OXOQ and [3H]ethyl-beta-carboline-3-carboxylate [( 3H]beta CCE) binding. The in vitro addition of GABA (10(-8)-10(-4) M) to spinal cord membrane preparations produced an increase in the binding of [3H]FNT and [3H]2OXOQ. The maximal enhancement produced by GABA was 50 and 82% above control values for [3H]FNT and [3H]2OXOQ, respectively. In contrast, GABA stimulated both [3H]FNT and [3H]2OXOQ binding in the cerebellum to a similar extent. We also evaluated the effects of different ligands for BZD recognition sites on the binding of [3H]GABA to spinal cord membranes, as compared with brain areas containing a higher proportion ( greater than 30%) of Type I sites. Diazepam, quazepam and the beta-carboline, ZK 93423, enhanced the specific binding of [3H]GABA in a concentration-dependent manner (10(-7)-10(-5) M) in the cerebral cortex and hippocampus but not in the spinal cord and cerebellum. These results indicate that there is a regional variation in the interaction between GABA and BZD recognition sites in the central nervous system. PMID- 2553448 TI - Chronic phenobarbital administration affects GABA and benzodiazepine receptors in the brain and periphery. AB - Chronic phenobarbital administration for 20 days to mice resulted in significant increases in the density of peripheral benzodiazepine binding sites in the heart, kidney and cerebellum, but did not affect the density of these sites in the olfactory bulb. No alteration in the affinity (KD) of peripheral benzodiazepine binding sites for their ligand [3H]PK 11195 was observed in any of the organs examined. Phenobarbital treatment did not affect the maximal binding capacity or the affinity of the central benzodiazepine receptors for [3H]flunitrazepam in the cerebral cortex, hippocampus and olfactory bulb. A significant reduction in [3H]muscimol binding in the hippocampus was obtained following chronic phenobarbital treatment. Such an alteration was not detected in the cerebral cortex. The KD values remained unaltered in both tissues. The modulatory effect of phenobarbital on peripheral benzodiazepine binding sites could be related to alterations in the functions of these organs or to the neurochemical effects of the drug. PMID- 2553449 TI - Noradrenaline and thyroid function regulate (Na+,K+)-adenosine triphosphatase independently in vivo. AB - We investigated interactions between noradrenaline and thyroid hormone status in the regulation of (Na+,K+)-ATPase in vivo. Treatment with the beta-adrenoceptor antagonist propranolol or with the neurotoxin 6-hydroxydopamine did not prevent the increases in heart (Na+,K+)-ATPase associated with triiodothyronine treatment. Administration of methimazole did not prevent the increase in (Na+,K+) ATPase indices in cerebral cortex and heart associated with subacute noradrenergic stimulation by yohimbine. There was no evidence for synergistic effects between thyroid hormone administration and noradrenergic stimulation by yohimbine. Thyroid hormone, unlike noradrenaline, mainly increased (Na+,K+) ATPase activity with low affinity for ouabain. These results show that noradrenaline and thyroid hormone regulate (Na+,K+)-ATPase by largely independent mechanisms, and may regulate different populations of enzyme molecules. PMID- 2553450 TI - Modulation of noradrenaline release in slices of rat kidney cortex through alpha 1- and alpha 2-adrenoceptors. AB - Slices of rat kidney cortex were incubated in [3H]noradrenaline, then placed in a flow cell and subjected to electrical field stimulation. At a stimulation frequency of 5 Hz, both the alpha 2-adrenoceptor antagonist idazoxan (0.1 microM) and the alpha 1-adrenoceptor antagonist prazosin (0.1 microM) significantly enhanced the stimulation-induced (S-I) outflow of radioactivity from the slice. However, neither clonidine (0.1 microM) nor methoxamine (10 microM), alpha 2- and alpha 1-agonists respectively, affected the S-I outflow of radioactivity at this stimulation frequency. At a lower stimulation frequency (1 Hz), the S-I outflow of radioactivity was not affected by idazoxan or prazosin, but was inhibited by both clonidine and methoxamine. The effect of clonidine was prevented by idazoxan (0.1 microM), but not by prazosin (0.1 microM). The effect of methoxamine was abolished by prazosin (0.1 microM), but not by idazoxan (0.1 microM). The inhibitory effect of methoxamine was not prevented by the prostaglandin synthesis inhibitor indomethacin (10 microM) or the adenosine receptor antagonist 8 phenyltheophylline (1 microM) and thus was not mediated by either prostaglandins or adenosine. The results suggest that both prejunctional alpha 1- and alpha 2 adrenoceptors are directly involved in modulation of noradrenaline release from the renal sympathetic nerves of the rat. PMID- 2553451 TI - [3H]WEB 2086 labels platelet activating factor receptors in guinea pig and human lung. AB - The radiolabelled platelet activating factor (PAF) receptor antagonist, [3H]WEB 2086, bound to specific sites on membrane fractions from homogenised guinea pig and human lungs. The sites on guinea pig and human membranes bound [3H]WEB 2086 with dissociation constants (KD) of 16.8 and 22.6 nM and binding capacities (Bmax) of 203 and 157 fmol/mg protein, respectively. In both species, binding was displaced competitively by PAF, suggesting that the sites labelled by [3H]WEB 2086 are PAF receptors. PMID- 2553452 TI - Susceptibility of four species of small rodents to encephalomyocarditis (EMC) virus infection. AB - The D variant of encephalomyocarditis virus (10(1)-10(5) PFU/head) was intraperitoneally inoculated into 4 species of small rodents, rats, mice, Syrian hamsters, and Mongolian gerbils, and the susceptibility of these animals to EMC virus was examined virologically and histopathologically 3 days after infection. Viral replication was detected in the brain (mice), in the heart (mice and gerbils), and in the pancreas (mice, hamsters, and gerbils). No viral replication was detected in rats. Histopathological changes were seen in the brain (mice and hamsters), in the heart (mice and gerbils), and in the pancreas (mice, hamsters, and gerbils). No histopathological changes were seen in rats. The present results suggest that it may be quite possible to produce EMC virus-induced diabetes mellitus not only in mice but also in hamsters and gerbils. PMID- 2553453 TI - Staurosporine-induced neurite outgrowth in PC12h cells. AB - Treatment of PC12h cells with staurosporine (100 nM), a potent inhibitor of protein kinases, promoted rapid outgrowth of neurites. The mechanism of neurite formation elicited by staurosporine is different from that elicited by nerve growth factor or by dibutyryl cyclic AMP, based on the independence from transcription or from activation of cyclic AMP-dependent protein kinase, respectively. Comparative experiments showed that of these three neurite promoting agents, staurosporine was the most effective in eliciting neurite initiation. PMID- 2553454 TI - Actin cytoskeleton of extraembryonic endoderm and teratocarcinoma-derived endoderm cells. AB - Distinct F-actin- and myosin-containing stress fibers were observed in situ in many endoderm cells of parietal yolk sacs from 11-day mouse embryos. In visceral endoderm (VE) such fibers were not seen, and F-actin was concentrated in the cell periphery. Correspondingly, in electron microscopy ventral cell membrane associated bundles of microfilaments were revealed in the periphery of parietal endoderm (PE) cells but not in VE cells. Both PE and VE cells formed stress fibers in primary cultures. Undifferentiated F9 embryonal carcinoma cells formed only short actin spikes and fibrils irrespective of growth substratum. In PE-like derivatives of F9 cells, on the other hand, distribution of F-actin was markedly affected by the growth substratum: They formed distinct stress fibers when plated on fibronectin but did not when plated on gelatin. Similarly, in teratocarcinoma derived PE cells (PYS-2) adhesion to fibronectin induced the formation of distinct bundles of F-actin and plaques of vinculin. The results suggest that the susceptibility of teratocarcinoma cell actin cytoskeleton to the influence of molecular composition of surrounding matrix is developmentally regulated. On the other hand, the reason for the presence of stress fibers in PE cells and for their absence in VE cells is unclear. PMID- 2553455 TI - Production of insulin-like growth factor binding proteins by small-cell lung cancer cell lines. AB - Conditioned serum-free media (CM) from small-cell lung cancer (SCLC) cell lines were examined for the presence of insulin-like growth-factor-binding proteins (IGF-BP). 6/9 SCLC cell lines secreted binding proteins with high affinity for IGFs. When [125I]IGF-I or [125I]IGF-II was incubated with the CMs, complexes of tracer with proteins could be demonstrated by gel filtration, by precipitation with polyethylenglycol, and after adsorption of unbound tracer with activated charcoal. Analysis of binding data according to the method of Scatchard resulted in linear plots for IGF-I and IGF-II. The dissociation constants were determined to be 0.106 nM for IGF-I and 0.209 nM for IGF-II binding. Cross-linking of [125I]IGF-I or [125I]IGF-II to the CMs followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under nonreducing conditions revealed the presence of IGF-BPs with molecular masses in the range 24-32 kDa. The binding was competitively inhibited by addition of cold IGF-I and IGF-II but not by insulin. Northern blot hybridization with an IGF-BP cDNA probe encoding a low-molecular-weight IGF-BP from a human placenta cDNA library and Western blot analysis with a corresponding polyclonal antibody showed no expression of this gene. These data demonstrate that SCLC cell lines release IGF-BPs in culture supernatants, which differ from IGF-BPs detected in liver and placenta. These IGF BPs might be important mediators in the autocrine/paracrine growth regulation of IGFs in SCLC. PMID- 2553456 TI - Human teratocarcinoma cells express functional insulin-like growth factor I receptors. AB - Using iodinated insulin-like growth factors (IGFs) we have detected receptors for IGF-I at the cell surface of the clonally derived human embryonal carcinoma cell line Tera 2 clone 13. Affinity crosslinking of IGFs to Tera 2 clone 13-derived membrane preparations revealed the presence of proteins with features of both type-I and type-II IGF receptors. Treatment of Tera 2 clone 13 cells with retinoic acid to induce differentiation results in an increased number of cell surface receptors, apparently without altering the ratio of type-I and type-II receptors. In addition, Tera 2 clone 13 IGF-I receptors catalyze (auto)phosphorylation at tyrosine upon IGF-I and insulin binding. These findings suggest that type-I IGF receptors might be involved in mediating the effects of IGFs and insulin upon the proliferation of Tera 2 clone 13 cells. PMID- 2553457 TI - Uptake of transcobalamin II-bound cobalamin by HL-60 cells: effects of differentiation induction. AB - Binding and uptake of transcobalamin II-bound cobalamin by HL-60 promyelocytic leukemia cells proceed through receptor-mediated endocytosis. The affinity constant of the receptor for transcobalamin II-cobalamin was found to be 6.1 liter/nmol and the maximal rate of uptake 12 pmol/10(9) cells/h. This uptake is mediated by about 3000 receptor sites per cell. Evidence is presented that the receptor recirculates from the cell surface to the lysosomes and vice versa. Upon differentiation induction of the cells by either DMSO in granulocytic direction or by 1,25-dihydroxy-vitamin D3 in monocytic direction a rapid decline in cellular uptake and cell surface binding of the protein-bound vitamin ensues. In particular the internalization of the complex decreases faster than all other observed signs of the ongoing differentiation process, such as reduction in the OKT9-reactive transferrin receptor, increase in lineage-specific surface markers, and decrease in [3H]thymidine incorporation and actual cell proliferation. The transcobalamin II receptor on the cell surface appears to be a proliferation associated membrane component in human leukemic cells. PMID- 2553458 TI - Rat mesangial cell-matrix interactions in culture. AB - The glomerular mesangium contains fibronectin (FN), laminin, and collagen IV, but it remains unclear whether these matrix proteins affect mesangial cellular functions. The present experiments were designed to test whether cell-matrix interactions could affect some functions of mesangial cells. Cultured rat mesangial cells synthesized a cellular form of FN that was both secreted and incorporated into an extensive, fibrillar pericellular matrix. This FN matrix was increased in high-density cultures and was more developed in human mesangial cells. Rat mesangial cells in vitro displayed a marked capacity to incorporate exogenous FN into a pericellular matrix, demonstrating that accumulations of FN in the mesangial matrix could result from endogenous and/or exogenous sources. Rat mesangial cells also expressed RGD-sensitive integrin receptors for FN, laminin, and collagens I and IV that promoted cell adhesion and that directed differential changes in morphology. Indirect evidence suggested the existence of other mesangial binding sites for extracellular matrix proteins. FN and collagen IV also stimulated modest increases in [3H]thymidine uptake and cell number by quiescent cells. Taken together, these results suggest that cultured mesangial cells present a model system for studying the regulation of cell-matrix interactions in the mesangium. PMID- 2553459 TI - Reversible extinction of insulin gene expression in insulinoma x fibroblast somatic cell hybrids. AB - To investigate for the presence of regulator(s) repressing the expression of insulin gene in cells other than pancreatic beta cells, rat insulinoma (RIN) cells secreting insulin were hybridized with fibroblasts from various species. In RIN x L mouse fibroblast hybrids, which maintained most of the parental chromosomes, no insulin transcripts were detected. In three RIN x Indian muntjac fibroblast hybrids and one RIN x human fibroblast hybrid which had lost DNA from the fibroblast parent through subculture, expression of the insulin gene was first extinguished and then reexpressed. This suggests that negative regulator(s), present in fibroblasts, can turn off insulin gene expression in RIN x fibroblast hybrids as long as the gene(s) contributed by the fibroblast are maintained. PMID- 2553460 TI - Interactions of human lymphoblasts with targeted vesicles containing Sendai virus envelope proteins. AB - We have studied the internalization of targeted fusogenic liposome content to leukemic T cells (CEM) in vitro. We describe a method for the covalent coupling of T101 antibody to the surface of liposomes and the incorporation of fusogenic viral protein into the liposome membrane. Hygromycin B, an impermeant inhibitor of protein synthesis, was encapsulated in the targeted fusogenic liposomes and delivered directly to the cytoplasm of leukemic T cells by fusion between the two membranes. The cytotoxic effect was measured by [3H]thymidine incorporation. We show that CEM are rapidly and specifically killed by the drug encapsulated in the targeted fusogenic liposomes. This effect is due to the binding of the liposome by means of the antibody and then to the fusion of the liposome with the targeted cell membrane, mediated by F protein. PMID- 2553461 TI - Mitogen-stimulated phosphorylation of nuclear proteins in Swiss 3T3 cells: evidence for a protein kinase C requirement. AB - When Swiss 3T3 fibroblasts are treated with a combination of IGF-I2 and bombesin at mitogenic concentrations, in vivo phosphorylation of some nuclear proteins occurs within 45-90 min. Among these proteins, histone H1 and a 0.75 M PCA soluble polypeptide with an apparent Mr of 21,000, as revealed by electrophoretic analysis, are phosphorylated in vitro by protein kinase C in isolated nuclei purified from 3T3 cells treated for 90 min with IGF-I and bombesin. Since these phosphorylative events follow the earlier changes, recently demonstrated, in nuclear polyphosphoinositide metabolism induced by the same mitogen combination, it seems possible that these two phenomena are related to each other and trigger the synthetic machinery responsible for replicating DNA. PMID- 2553462 TI - Identification of a high-affinity Ca2+ pump associated with endocytotic vesicles in Dictyostelium discoideum. AB - In the cellular slime mold Dictyostelium discoideum, changes in free cytosolic Ca2+ are thought to regulate certain processes during cell aggregation and differentiation. To understand the mechanisms controlling free Ca2+ levels in this organism, we previously isolated and characterized an ATP/Mg2+-dependent, high-affinity Ca2+ pump which appeared to be a component of "inside-out" plasma membrane vesicles [J. L. Milne and M. B. Coukell (1988) Biochem. J. 249. 223 230]. In this report, we demonstrate that a high-affinity Ca2+ pump, with properties virtually identical to the isolated pump, can be detected in filipin- or digitonin-permeabilized cells of Dictyostelium. Moreover, Ca2+-pumping vesicles, which migrate on Percoll/KCl gradients like the vesicles identified earlier, can be isolated from the permeabilized cells. Results of additional experiments suggest that this intracellular Ca2+ transporter is associated with a high-capacity non-IP3-releasable Ca2+ store which is generated by endocytosis. A possible role for this store in maintaining Ca2+ homeostasis in Dictyostelium is discussed. PMID- 2553463 TI - Increase in c-fos and c-myc mRNA levels in untransformed and SV40-transformed 3Y1 fibroblasts after addition of serum: its relationship to the control of initiation of S phase. AB - When rat 3Y1 fibroblasts were exposed to serum after 7.5 h of S, G2, and M phases in the absence of serum, the c-fos and c-myc mRNA levels markedly increased. This marked increase was also observed when density-arrested cells were stimulated with fresh serum to initiate proliferation. Increase in the c-fos and c-myc mRNA levels was not observed in cells that had traversed 7.5 h in these phases in the presence of serum. Cells passing through S, G2, and M phases in the absence of serum delayed entry into the next S phase approximately 8 h compared to control cells incubated in the presence of serum. Also, when density-arrested cells were stimulated with serum for 5 h, then deprived of serum for 8 h, and then incubated in serum again, the c-fos and c-myc mRNA levels increased. In this last case, the total excess time of serum exposure required to enter S phase was only 2 h, indicating that cells had not returned to the initial density-arrested state during the serum deprivation period. The increase in c-fos and c-myc mRNA levels following addition of serum after incubation in the absence of serum was also observed in SV40-transformed 3Y1 cells. The entry of SV40-transformed cells into S phase was not markedly affected by the absence of serum. These results can be explained by assuming that there is a process leading to the initiation of S phase that is operating or accumulating continuously in all cell cycle phases. In 3Y1 cells the expression of the c-fos and c-myc genes is required at any cell cycle phase, and the increase in c-fos and c-myc mRNA levels in response to changes in serum concentration simply reflects the possible overexpression due to the delay of a hypothesized negative feedback regulation. In SV40-transformed 3Y1 cells, the process leading to the initiation of S phase operates normally in response to growth factors, and the SV40 large T antigen supplements or enhances the process in the absence of the growth factors. PMID- 2553464 TI - Concurrent outbreaks of influenza and parvovirus B19 in a boys' boarding school. AB - In the spring term of 1985 there was a protracted outbreak of upper respiratory tract febrile illness consistent with a clinical diagnosis of influenza in a boys' boarding school, which lasted from 23 January to 29 March. Although influenza virus infection was confirmed in 89% of cases in the first half of the term, 53% of the cases which occurred in the second half of the term had no evidence of infection with influenza virus. Between 5 February and 31 March 28 boys presented with skin rashes consistent with a clinical diagnosis of erythema infectiosum; 68% of these were associated with parvovirus B19. Investigation of the cases of clinical influenza with no identified respiratory pathogen revealed a 58% infection rate with B19. B19 DNA was identified in either throat swabs or acute stage bloods of nine pupils with influenza-like symptoms. Cohort studies revealed that 44% of pupils aged 15-16 years were immune before the outbreak compared with only 17% of pupils aged 11-12 years. Infection in the younger group was common and was associated with influenza-like illnesses as well as rashes. Forty-eight per cent of those who did not report any symptoms were also infected with B19. PMID- 2553465 TI - Investigation of an outbreak of hepatitis A simplified by salivary antibody testing. AB - In March 1988 a general practitioner notified two cases of hepatitis A in a private boarding school. Epidemiological investigation, including testing for salivary antibodies revealed a further five cases and established immunity to, and recent infection with, hepatitis A virus (HAV). The pattern of the outbreak was described. A number of practices which would encourage cross-infection were corrected. Normal human immunoglobulin was given to contacts. Repeat salivary testing 10 weeks later revealed that two more boys had become reactive for anti HAV, though at a low titre. These may have been serological responses to HAV infection modified by the passive immunization. PMID- 2553466 TI - Hepatitis A antibody in blood donors in North East Thames region: implications to prevention policies. AB - A total of 1786 blood donors were screened for the presence of anti-hepatitis A antibody (anti HAV). 64.5% of the donors were found to be positive. The prevalence of the antibody was found to be age-related, 55% at 18 years and 75% at 65 years. No relationship was noted between the presence of antibody, foreign travel or a specific destination. Assay of antibody levels in selected seropositive individuals gave a mean level of 5.0 IU/ml. The prevalence of infection in this selected population is important in the context of passive immunization with normal human immunoglobulin and for defining a policy of immunization with hepatitis A vaccines, which are currently undergoing clinical trials. PMID- 2553467 TI - Toward a dietary prevention of cancer. AB - Clearly, there is a need for further elucidation of the role in carcinogenesis of dietary factors. On the epidemiologic level, studies of the last few years indicate the need to investigate further the potential role of fats as related to all the major sites of cancer, to distinguish the effects of various fractions of fiber as well as of fibers deriving from fruits, vegetables and grains, and to examine more closely carotenoids as opposed to retinoids to explore further the proposition that although carotenoids may reduce risk for a number of cancers, retinoids may play no part. Future studies must also reflect more carefully the human experience regarding diet. Because humans do not ingest foods singly, but rather as groups, patterns of intake of groups of foods and nutrients may provide insight into types of diet that affect risk. In nutritional science, there is an increasing appreciation of the complexity of interaction of nutrients both in absorption and in metabolism; future epidemiologic studies of biologic materials as well as of nutrient interactions may be beneficial. Interindividual differences in response to diet which may identify subgroups who are more or less susceptible to particular dietary factors should also be examined. Since dietary exposures affect virtually everyone, the public health implications of even small differences in risk are important. For example, a well substantiated relative risk of 1.2 for a nutrient associated with breast cancer is not trivial. Speculation as to the proportion of total cancer attributable to diet is so tenuous as to be almost frivolous.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553468 TI - Corticosteroid action on choroid plexus: reduction in Na+-K+-ATPase activity, choline transport capacity, and rate of CSF formation. AB - Glucocorticoids have a well-known clinical effect on brain edema and intracranial hypertension, but the mechanism of action is still poorly understood. In the present report the effect of beta-methasone on choroid plexus transport and CSF formation was studied. Following 5 days of daily treatment with betamethasone the CSF production rate in rabbits was reduced by 43% as measured by ventriculo cisternal perfusion with radioactive inulin. Accordingly, the transport capacity in the choroid plexus, measured in terms of choline uptake and accumulation in vitro, and the activity of Na+--K+-ATPase decreased in both rabbit (in the lateral ventricles by 31 and 31%, respectively) and rat (by 16 and 24%, respectively). Thus, the demonstrated influence of glucocorticoids on these functions of the choroid plexus seem to be important components in their therapeutic effect on intracranial hypertension. PMID- 2553469 TI - Modification of evoked hippocampal dentate inhibition by diazepam and three antagonists in urethane-anesthetized rats. AB - Urethane-anesthetized rats with perforant pathway stimulating electrodes and recording electrodes placed in the hippocampal dentate gyrus were exposed to increasing doses of either the benzodiazepine agonist diazepam or an antagonist (PK-11195, CGS-8216, and RO15-1788). Analysis of the monosynaptic evoked field potentials indicated that none of the four compounds altered the threshold for eliciting the excitatory postsynaptic potential (EPSP). Reductions in field population spike (PS) amplitudes were seen after exposure to RO15-1788, CGS-8216, and diazepam, but not PK-11195. Using a paired-pulse paradigm, diazepam significantly increased early gamma-aminobutyric acid (GA-BAA)-mediated recurrent inhibition. The antagonist RO15-1788, but not CGS-8216 or PK-11195, also significantly increased early GABAA-mediated inhibition. The increase in GABAA mediated inhibition after diazepam was reversed by the subsequent administration of the central antagonists RO15-1788 or CGS-8216, but not the peripheral antagonist PK-11195. Pretreatment with CGS-8216 or RO15-1788 prevented diazepam induced inhibition. These data support the important modulatory role of the central benzodiazepine receptor in early GABAA-mediated inhibition at this synapse. They also suggest that basal granule cell excitability is not importantly modulated by this benzodiazepine receptor. PMID- 2553470 TI - Anticonvulsant actions of MK-801 on the lithium-pilocarpine model of status epilepticus in rats. AB - MK-801, a noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist, was tested for anticonvulsant effects in rats using two seizure models, coadministration of lithium and pilocarpine and administration of a high dose of pilocarpine alone. Three major results are reported. First, pretreatment with MK 801 produced an effective and dose-dependent anticonvulsant action with the lithium-pilocarpine model but not with rats treated with pilocarpine alone, suggesting that different biochemical mechanisms control seizures in these two models. Second, the anticonvulsant effect of MK-801 in the lithium-pilocarpine model only occurred after initial periods of seizure activity. This observation is suggested to be an in vivo demonstration of the conclusion derived from in vitro experiments that MK-801 binding requires agonist-induced opening of the channel sites of the NMDA receptor. Third, although it is relatively easy to block seizures induced by lithium and pilocarpine by administration of anticonvulsants prior to pilocarpine, it is more difficult to terminate ongoing status epilepticus and block the lethality of the seizures. Administration of MK 801 30 or 60 min after pilocarpine, i.e., during status epilepticus, gradually reduced electrical and behavioral seizure activity and greatly enhanced the survival rate. These results suggest that activation of NMDA receptors plays an important role in status epilepticus and brain damage in the lithium-pilocarpine model. This was further supported by results showing that nonconvulsive doses of NMDA and pilocarpine were synergistic, resulting in status epilepticus and subsequent mortality. PMID- 2553472 TI - Depressor effects of muscarinic and non-muscarinic mediation induced by lateral hypothalamic stimulation in the cat. AB - Transient sympathetically-mediated depressor effects were induced by stimulation of a small locus in the lateral hypothalamic peri-fornical region, medial to the fields of Forel. The ganglionic blocking agent, atropine methyl nitrate (ATMN), was used to show that muscarinic as well as non-muscarinic sympathetic ganglion receptor neurotransmission was involved. Evidence is presented that stimulation of this LH site co-activates a number of mechanisms and that depending on which of these are activated, the ganglionic blocking agent ATMN may either block, reverse or potentiate the depressor effect. PMID- 2553471 TI - Central melatonin receptors: implications for a mode of action. AB - The influence of melatonin on circadian and photoperiodic functions in numerous species is well documented. It is known that the effect of melatonin on circadian rhythmicity is mediated via the suprachiasmatic nucleus (SCN), the biological clock of the brain. It is not known however where the photoperiodic effects of melatonin are mediated. Evidence from brain lesioning and melatonin implant studies point to a site in or near the medial hypothalamus. In contrast to these studies, melatonin receptors have been reported in widespread areas of the brain, the pituitary and in peripheral tissues. The characteristics of the reported melatonin receptors vary widely between studies and consequently no definitive description of a physiologically relevant melatonin receptor has received universal recognition. This review marshals recent evidence for the localization and characterization of the melatonin receptor and discusses these findings in the context of the known effects of the hormone in different species. PMID- 2553473 TI - Expression of A-type PDGF receptor in cytoplasm of tumor cell lines synthesizing PDGF. AB - Two tumor cell lines, WM 266-4 melanoma and SW 707 colorectal carcinoma, both of which synthesize platelet-derived growth factor (PDGF) but do not express cell surface PDGF receptor, were tested for the presence of intracytoplasmic PDGF receptor. Using a novel method of intracytoplasmic (newly synthesized), receptor precipitation by the growth factor, we identified a 125,000 molecular weight protein crosslinked with 125I-labeled PDGF in a 140,000 molecular weight complex. In vitro translation of the corresponding mRNA revealed a 125,000 molecular weight product which presumably represents the A-type PDGF receptor. Exogenous PDGF was found to activate synthesis of RNA and cell proliferation. PMID- 2553474 TI - Development of a murine model to study the pathogenesis of rotavirus infection. AB - A murine model to study enteritis induced by bovine (BRV) and murine rotavirus (MRV) has been developed. The course of infection was determined by clinical symptoms of diarrhea and virus isolation as well as histopathological, immunohistochemical, and electron microscopic methods. Both isolates were able to replicate and produce clinical symptoms in neonatal mice. Rotavirus-free neonates were orally inoculated with MRV or BRV and observed over a 192-hr postinoculation (HPI) period. Following infection with 10(4) PFU of virus, diarrhea and maximal intestinal dysfunction, as measured by xylose absorption, did not occur until beyond 20 hr postinfection even though maximal virus production occurred at 10-15 HPI. Immunohistochemically and by electron microscopy we were able to demonstrate viral antigen and virus particles in the enterocytes of villous tips at 5-8 HPI. The appearance of diarrheal symptoms was dependent on the virus dose and the type of virus isolate inoculated. The disease could be induced with doses as low as 1 x 10(2) PFU/mouse of BRV and 1 x 10(1) PFU/mouse of MRV. On the basis of these results, MRV was found to be more virulent than BRV in this model. The model should prove useful for studies designed to assess rotavirus virulence genes and for vaccine protection studies. This work emphasizes the need for early sample collection for critical evaluation of any vaccine or antiviral agent using this model. PMID- 2553475 TI - Tumour markers in the clinical management of patients with small cell lung cancer. PMID- 2553476 TI - Combined measurements of neuron specific enolase and bombesin/gastrin releasing peptide in lung cancer. AB - Pretreatment serum neuron specific enolase (NSE) and plasma bombesin/gastrin releasing peptide (BN/GRP) were measured in 92 lung cancer patients and 17 controls. The mean level of NSE (p less than 0.001) and BN/GRP (p less than 0.05) was significantly raised in patients with small cell lung cancer (SCLC, n = 62) compared to non-SCLC (n = 30) and controls. The mean concentration of NSE in extensive SCLC was significantly greater (p less than 0.005) than in limited stage but with a substantial overlap of values. Forty-seven out of 62 SCLC patients had at least one of the two markers raised (sensitivity 76%, specificity 83%), 44 had raised NSE (sensitivity 71%, specificity 89%) but only 24 had BN/GRP raised (sensitivity 42%, specificity 91%). At restaging, 16 of 19 patients with SCLC responsive to chemotherapy showed a significant fall of NSE; on the other hand, BN/GRP fell significantly in only 3 patients, remaining unchanged in the majority of responding patients. In conclusion, the combined determination of NSE and BN/GRP in SCLC, at diagnosis and during the follow-up, was not found to be superior to NSE determination alone. PMID- 2553477 TI - [Effect of several m-cholinoblockaders on the functioning of sodium potential dependent channels of neuroblastoma cells]. AB - Effects of two central antimuscarinic drugs, namely N-methyl-4-piperidinol benzylate and tropine phenylcyclopentylglycolate on Na currents in internally dialyzed mouse neuroblastoma cells were studied using the suction-pipette voltage clamp technique. Like amine local anesthetics, the both compounds produce two phenomenologically different types of sodium current inhibition: a steady block without conditioning and a cumulative (use-dependent) block which develops during repetitive membrane pulsing. The ability of these m-cholinoblockers to modulate voltage-dependent ionic permeability of the neuronal membranes is supposed to be one of possible components in the mechanism of their both main and accessory (non synaptic) effects. PMID- 2553478 TI - Functional characteristics of calcitonin gene-related peptide receptors in human Ewing's sarcoma WE-68 cells. AB - Calcitonin gene-related peptide (CGRP) receptor activity was studied in WE-68 human Ewing's sarcoma cells. 125I-human CGRP bound in a time-dependent, reversible and saturable manner. Scatchard plots were compatible with the presence of a homogenous population of CGRP receptors with high affinity (Kd = 15 pM, and Bmax = 1.9 fmol/mg protein). The potency order of unlabeled peptides in the presence of radioligand, was: human CGRP-II greater than human CGRP = chick CGRP greater than rat CGRP = rat [Tyr0]CGRP greater than human [Tyr0] CGRP much greater than salmon calcitonin (CT) greater than rat [Tyr0]CGRP-(28-37). Each peptide except CT and [Tyr0]CGRP-(28-37) stimulated cyclic AMP generation in a concentration-dependent manner, and the relative potencies paralleled their relative ability in inhibiting 125I-human CGRP binding. We conclude that WE-68 Ewing's sarcoma cells express genuine CGRP receptors which upon activation lead to stimulation of cyclic AMP formation PMID- 2553479 TI - EPR characterization of soluble fragments of succinate dehydrogenase from mutant strains of Bacillus subtilis. AB - Succinate dehydrogenase is a membrane-bound metallo-flavo-enzyme containing a bi- (S-1), a tri- (S-3) and a tetranuclear (S-2) iron-sulfur cluster. The catalytic portion of the enzyme contains two distinct subunits designated Fp and Ip. Using concentrated extracts from mutant strains of Bacillus subtilis it was demonstrated, by using low temperature EPR, that cluster S-2 can be assembled in a soluble succinate dehydrogenase. In a mutant with a truncated Ip subunit which lacks 7 of the 11 conserved cysteine residues, cluster S-1 lacked the spin relaxation properties attributable to an adjacent cluster S-2. These data are consistent with a model where one or more cysteine residues from the middle set of 4 conserved cysteines in the Ip subunit are ligands to the tetranuclear cluster. PMID- 2553480 TI - Calcium mobilization and Na+/H+ antiport activation by endothelin in human skin fibroblasts. AB - Endothelin (ET-1) has been shown to exert vasoconstrictor activity in vivo and mobilize Ca2+ in vascular smooth muscle cells in culture. In this paper we show that the human skin fibroblast exhibits specific receptors to ET-1 and that activation of these receptors results in increased intracellular Ca2+ (Ca2+i) and accelerated Na+/H+ antiport activity. ET-1 raised Ca2+i in a dose-response manner; the peak Ca2+i rise was from basal levels of 112.2 +/- 21.9 to 299.2 +/- 49.7 nM at 300 nM ET-1. This rise was attenuated by removal of extracellular Ca2+i0. Although ET-1 did not alter basal intracellular pH, it enhanced Na+/H+ antiport activity of acidified cells. Fibroblasts demonstrated 156 +/- 18 (mean +/- SE) ET-1 receptors per unit cell and an equilibrium dissociation constant of 203.4 +/- 35.6 pM. Inasmuch as ET-1 plays a role in the metabolism of cells such as the undifferentiated fibroblast, an important action of this peptide may be to act as a growth factor. PMID- 2553481 TI - Retinoic acid regulation of the expression of retinoic acid receptors in wild type and mutant embryonal carcinoma cells. AB - The embryonal carcinoma cell line P19 undergoes terminal differentiation upon induction by retinoic acid (RA). The effects of RA on the retinoic acid receptor (RAR) mRNA level were investigated in P19 cells and an RA non-responsive mutant RAC65. RA induced a rapid accumulation of RAR alpha within 2 h and this was followed by an increase in the RAR beta mRNA. RAC65 cells differed from P19 cells in that the accumulation of RAR alpha mRNA was delayed and that an aberrant pattern of RAR alpha transcript was observed. PMID- 2553482 TI - Advances in Na+,K+-ATPase studies: from protein to gene and back to protein. AB - Complete primary structures of both subunits of Na+,K+-ATPase from various sources have been established by a combination of the methods for molecular cloning and protein chemistry. The gene family homologous to the alpha-subunit cDNA of animal Na+,K+-ATPases has been found in the human genome. Some genes of this family encode the known isoforms (alpha I and alpha II) of the Na+,K+-ATPase catalytic subunit. The proteins coded by other genes can be either new isoforms of the Na+,K+-ATPase catalytic subunit or other ion-transporting ATPases. Expression of the genes of this family proceeds in a tissue-specific manner and changes during the postnatal development and neoplastic transformation. The complete exon-intron structure of one of the genes of this family has been established. This gene codes for the form of the catalytic subunit, the existence of which has been unknown. Apparently, all the genes of the discovered family have a similar intron-exon structure. There is certain correlation between the gene structure and the proposed domain arrangement of the alpha-subunit. The results obtained have become the basis for the experiments which prove the existence of the earlier unknown alpha III isoform of the Na+,K+-ATPase catalytic subunit and have made possible the study of its function. PMID- 2553483 TI - Prolonged ischemia differently affects phospholipase C acting against phosphatidylinositol and phosphatidylinositol 4,5-bisphosphate in brain subsynaptosomal fraction. AB - The effect of 10 min ischemia on the activity of phospholipase C acting against [3H]inositol-phosphatidylinositol (PI) and [3H]inositol-phosphatidylinositol 4,5 bisphosphate (PIP2) in the brain subsynaptosomal fractions was investigated. In the presence of endogenous CaCl2, specific activity of phospholipase C acting on phosphatidylinositol was as follows: synaptic cytosol (SC) greater than synaptic vesicles (SV) greater than synaptic plasma membrane SPM). Brain ischemia activated phospholipase C acting on PI by about 60% and 40% in SV and SPM, respectively. The enzyme of synaptic cytosol was not affected by ischemic insult. Phospholipase C acting against PIP2 in the presence of endogenous calcium expressed the specific activity in the following order: SV greater than SPM greater than SC. After 10 min of brain ischemia, activity of phospholipase C acting on PIP2 was significantly suppressed in all subsynaptosomal fractions by about 50-60%. These results indicate that prolonged ischemia produced activation exclusively of phospholipase C acting against phosphatidylinositol. PMID- 2553484 TI - High resolution 1H NMR study of the solution structure of the S4 segment of the sodium channel protein. AB - A peptide (S4) of the rat brain sodium channel has been synthesized, studied by high-resolution NMR and its secondary structure modelled by distance geometry and restrained molecular dynamics techniques. PMID- 2553485 TI - Addition of ATP increases the apparent molecular mass of the multicatalytic proteinase, ingensin. AB - A high-molecular mass ATP-dependent proteinase was shown to be identical to a multicatalytic proteinase, ingensin [(1988) Eur. J. Biochem. 177, 261-266]. The molecular mass of this proteinase increased in crude extracts of the rat liver and porcine brain, but not in the purified sample, only when the proteinase was extracted with ATP. The higher-molecular form of ingensin may be the intact one, because the concentration of ATP in vivo never decreases below 1 mM. This form of the proteinase is latent and it requires a high concentration of detergent for activation. On chromatography, it was found that the high-molecular form corresponds to the previously reported minor isoenzyme of ingensin [(1986) Biochim. Biophys. Acta 882, 297-304], ingensin A, or possibly to the ATP/ubiquitin-dependent 26S protease [(1987) J. Biol. Chem. 262, 8303-8313], and the low-molecular form to major ingensin B or the ATP/ubiquitin-independent 20 S protease. PMID- 2553486 TI - Influence of surface charge on the incorporation and orientation of cytochrome c oxidase in liposomes. AB - Cytochrome c-oxidase is usually oriented 80-90% right-side-out when reconstituted with asolectin by the cholate dialysis method. Transformation of positively charged lysine groups at the matrix domain into negatively charged groups with succinic anhydride results in random orientation. A random orientation is also found after reconstitution in phosphatidylcholine, which can be changed into predominant right-side-out orientation by addition of cardiolipin. It is concluded that electrostatic interaction between positively charged groups of cytochrome c-oxidase with negative groups of phospholipids determines the asymmetric orientation of the enzyme in liposomes. PMID- 2553487 TI - A 1H NMR study of bovine cytochrome oxidase. Paramagnetically shifted resonances of haem a. AB - Dimeric and monomeric forms of mitochondrial cytochrome oxidase (EC 1.9.3.1) have been examined using 1H NMR spectroscopy. Paramagnetically shifted resonances were detected in spectra of the monomeric protein. Studies of this protein in a number of oxidation and ligation states have assigned these resonances to ferrihaem a. The temperature and pH dependence of this new probe of haem a environment is reported. PMID- 2553488 TI - The O2.- generating oxidase activation of bovine neutrophils. Evidence for synergism of multiple cytosolic factors in a cell-free system. AB - Activation of the O2.- generating oxidase in neutrophils can be achieved with a cell-free oxidase-activating system, which consists of a high speed supernatant (cytosol), a particulate fraction enriched in plasma membrane, GTP-gamma-S, arachidonic acid and Mg ions. Cytosolic proteins from bovine neutrophils were fractionated by chromatography on Mono Q and Mono S columns into two fractions, neither of which was able to activate efficiently the membrane-bound oxidase. However, when combined and added to the cell-free system under optimized conditions, they acted synergistically, activating the oxidase to virtually the same extent as crude cytosol. This synergism demonstrates that more than one cytosolic factor is required for oxidase activation, and can be used to trace the cytosolic factors during the course of their purification. PMID- 2553489 TI - N-formyl-methionyl-leucyl-phenylalanine induced accumulation of inositol phosphates indicates the presence of oligopeptide chemoattractant receptors on circulating human lymphocytes. AB - Using [3H]-inositol-labeled human lymphocytes, formation of inositolphosphates was found as a specific response to the chemotactic formylpeptide, N-formyl methionyl-leucyl-phenylalanine (fMLP), in a fashion similar to the effects of fMLP in human granulocytes. Inositol phosphate formation could be significantly augmented by lithium ions. The results suggest that fMLP-mediated hydrolysis of phosphoinositides is involved in its chemotactic effects on human lymphocytes. PMID- 2553490 TI - Molecular cloning of the Na,K-ATPase alpha-subunit in developing brine shrimp and sequence comparison with higher organisms. AB - We report here the molecular cloning, nucleotide sequence, and predicted amino acid sequence of an alpha-subunit of the developmentally useful model, Artemia. The amino acid sequence shows divergence from that of mammals, birds, Torpedo, and Drosophila. However, regions in the putative ATP binding and transmembrane domains show absolute or high levels of conservation. Major differences occur in the amino-terminal domain and several other hypervariable regions. These differences are consistent with the suggestion that the brine shrimp is a 'fast clock' organism which diverged from the precursors of vertebrates 0.5-1 billion years ago. PMID- 2553491 TI - Granulocyte/macrophage colony-stimulating factor primes human neutrophils for increased diacylglycerol generation in response to chemoattractant. AB - Pretreatment of human neutrophils with granulocyte macrophage-colony stimulating factor (GM-CSF) augments several biological responses to chemoattractants (e.g. the respiratory burst, degranulation, and chemotaxis). However, little is known regarding the intracellular effects of priming with GM-CSF. In the present study, we have investigated the effects of GM-CSF on the generation of diacylglycerol (DAG), a proposed mediator of neutrophil responses. GM-CSF alone produced only a small increase in cellular DAG mass, which was most apparent after 30 min. GM-CSF pretreatment (60 min), however, caused a striking augmentation in DAG generation in response to the chemoattractant formyl-methionyl-leucyl-phenylalanine (fMLP), compared with neutrophils preincubated without GM-CSF. The augmentation in DAG generation correlated with an enhancement by GM-CSF of superoxide generation in response to fMLP. The data suggest that GM-CSF may exert some of its biological effects by enhancing DAG generation in response to a second agonist. PMID- 2553492 TI - Immunoaffinity isolation of Na,K-ATPase alpha 3 isoform from pig kidney. AB - The Na,K-ATPase alpha 3 isoform of the catalytic subunit has been isolated from pig kidney microsomes. The procedure employs immunoaffinity chromatography on Sepharose 4B covalently coupled with monospecific antibodies a-II against the synthetic peptide including the putative alpha 3 N-terminus. The structural analysis provides unambiguous proof that the isolated protein corresponds to the third transcript for the alpha 3 isoform. The N-terminal amino acid sequence determined. Met-Gly-Asp-Lys-Lys-Asp-Asp, shows that unlike the alpha 1 and alpha 2 proteins, the mature Na,K-ATPase isoform lacks post-translational proteolytic processing. PMID- 2553493 TI - Cyclic nucleotides and GTP analogues stimulate light-induced phosphorylation of octopus rhodopsin. AB - Light-induced phosphorylation of octopus rhodopsin in microvillar membrane was shown to be stimulated by cyclic nucleotides in contrast to vertebrate rhodopsin kinase. Non-hydrolyzable GTP analogues, GTP lambda S and GppNHp, greatly enhanced the light-induced phosphorylation of octopus rhodopsin, but the non-hydrolyzable GDP analogue, GDP beta S, was not effective. These results suggest that rhodopsin A-kinase is involved in regulating the interaction between rhodopsin and G protein in octopus photoreceptors. PMID- 2553494 TI - In vitro interaction of mutagenic chromium (VI) with red blood cells. AB - The interaction of mutagenic Cr(VI) with red blood cells has been studied by ESR spectroscopy. Signals of two Cr(V) species are observed almost immediately after contacting red cells with chromate(VI) aqueous solution at pH 7.4. The signal at go = 1.985, which decays within one hour, is attributed to a Cr(V) complex formed by glutathione due its reducing and chelating ability. The other signal at go = 1.979, which is distinctly more persistent, may indicate that some immobilization of the formed Cr(V) ions takes place on the macromolecular cell components, e.g. glycoproteins. PMID- 2553495 TI - In vivo activity of the most proximal promoter of the human aldolase A gene and analysis of transcriptional control elements. AB - The genomic region upstream from exon F (exon IV) of the human aldolase A gene has been studied for its ability to direct the transcription of a reporter gene in vivo. Transfection experiments in human hepatoma cells (Hep 3B) followed by CAT assay, and S1 mapping analysis, demonstrated that: (i) this region is able to drive CAT gene transcription; (ii) all the transcriptional control elements of this promoter are downstream from nucleotide -384 of the longer ubiquitous RNA start site and the sequences between -384 and -262 play a crucial role in transcriptional efficiency; (iii) initiation starting points for two mRNAs exist 61 bp apart. Gel retardation and footprinting assays demonstrated the presence of DNA-protein complexes mainly in the region between -384 and -262 and such ubiquitous binding factors as Sp1 and AP-1. PMID- 2553496 TI - Spatial and temporal expression pattern of retinoic acid receptor genes during mouse bone development. AB - Spatial and temporal expression pattern of retinoic acid receptor (RAR) genes was investigated in mouse finger bones during development by an in situ hybridization method with riboprobes synthesized from a human cDNA of the RAR-alpha. We found that the RAR genes are expressed intensively and specifically in calcifying fronts of the mouse finger bones, whereas the expression pattern is rather uniform in the limb buds and cartilage matrices of the embryonic fingers. Our findings are consistent with the fact that vitamin A is essential for normal mammalian bone development. PMID- 2553497 TI - A nursing challenge: cytomegalovirus in the transplant recipient. AB - Primary or reactivated CMV infection can be severe and life-threatening in the immunosuppressed transplant recipient. Nurses must be knowledgeable of the complex relationship between the infection and the compromised host state. By recognizing that all transplant recipients are susceptible to CMV infection, the nurse can prepare to care for these patients in the most effective manner. PMID- 2553498 TI - [The role of the brain monoaminergic systems in the effects of tuftsin and its analog on animal emotional behavior]. AB - The effects of tuftsin and its analogue TP-1 (300 micrograms/kg) on the animal adaptive behaviour in emotionally different situations, the level of biogenic amines and their metabolites 10, 30 min., 2 and 6 hrs after injection of these substances were studies. The TP-1 induced an increase in the memory traces stability better than tuftsin in emotionally negative situation. A single injection of the peptides induced changes in NA, DA and their metabolites contents for 30 min. to 2 hrs. In contrast to tuftsin, the TP-1 activated the metabolism of 5-HT in the hypothalamus and caudal brainstem without changing the NA turnover. The data obtained suggests that a stronger psychotropic activity of the TP-1 seems to be caused by its obvious effect on biogenic amines and their metabolites level in the brain. PMID- 2553499 TI - [Increase in the contractile force of the skeletal muscles during the reflex activation of the sympathetic nervous system]. AB - In anesthetized cats, the muscles of hindlimb if contracting in steady-state regimen (one brief tetanic contraction every 5 sec) under direct stimulation with trains of electrical pulses increase their contraction force following a sudden decrease of arterial blood pressure (down to 60 mm Hg) or asphyxia. This reflex reaction reaches its maximum (the increase in the force by 38.2 +/- 6.6%) if the muscle contracts with 25% of the maximal force. The sympathetic nerve influence is transduced through beta adrenoreceptors. PMID- 2553500 TI - [Insulin regulation of 2 kinetically different types of calcium current]. AB - At least two kinetically different kinds of calcium current are shown to exist in the frog atrial cells. The current with faster activation kinetics is usually depressed by insulin. Insulin also increases the amplitude of the slower calcium current. Pretreatment of atrial cells with cycloheximide does not change the effect of insulin on the fast calcium current but dramatically facilitates the insulin-induced activation of the slower calcium current. PMID- 2553501 TI - [Quantitative evaluation of the function of the presynaptic apparatus of the rat in cold adaptation]. AB - Binomial parameters of transmitter secretion were calculated on the basis of synaptic potential analysis for the diaphragm and extensor digitorum longus muscles of the control and cold-adapted rats. The quantum content, amplitude of EPPs and the amount of available transmitter increased in these muscles of cold adapted rats. This may be due to the influences of neurohumoral factors on the presynapse or, on the other hand, to changes in metabolism of the nerve terminal. Differences in presynaptic characteristics of the muscles between the control and the cold-adapted rats indicate the high reliability of the neuro-muscular junction functioning during slow rhythmic stimulation in cold ambient condition and the identical processes in the nerve terminal in the two groups of muscles. PMID- 2553502 TI - [Effect of the hypophyseo-adrenal cortical system on the vasopressinergic system]. AB - The effects of stress, ACTH and hydrocortisone treatment on the blood arginine vasopressin were investigated in rats. The plasma basal level of vasopressin decreased in response to administration of hydrocortisone (300 mg/100 g, i.p.). ACTH (0.1 U/100 g., i.p.) had no effects in the basal level of plasma vasopressin in adrenalectomized rats. Hydrocortisone administration resulted in an increase of plasma vasopressin in response to immobilization stress. The stress resulted in an initial decrease of plasma vasopressin followed by a vasopressin concentration increasing up to basal level. PMID- 2553503 TI - [Correlation between the levels of catecholamines (noradrenaline, adrenaline) and adrenal steroids (DHA-S, cortisol) in maternal and fetal blood during pregnancy and labor]. AB - It is known that both catecholamines (CA) and cortisol (F) levels elevate during labor. To determine the correlation between adrenal steroids and medullary function, maternal blood was collected during pregnancy, first stage of labor (MVI) and at delivery (MVII). Umbilical arterial and venous blood (UA, UV) was also obtained at delivery. Further, ACTH or dexamethasone (Dx) was given during the first stage of labor, and maternal blood was collected before and 30 minutes after the administration. Plasma levels of CA[noradrenaline (NA), adrenaline (Ad)] were extracted by trihydroxyindole method and were measured by HPLC. DHA-S and F levels were determined by specific RIA. Results are as follows: 1. No apparent change was observed in maternal NA and Ad levels throughout pregnancy. DHA-S levels were high in first trimester and decreased as pregnancy advanced, while F levels showed an increase trend as pregnancy progressed. 2. All hormone levels in maternal blood increased remarkably during labor. A significant negative correlation between F and Ad levels at delivery was noted. When F levels were elevated by ACTH administration, Ad levels decreased. Ad levels elevated when F levels were suppressed by Dx administration. 3. NA, Ad and DHA-S levels in cord blood were higher than those in MVII. Levels of F in maternal blood were higher than those in cord blood. A significant correlation of F in MVII and UA was observed. These results indicate that the suppressive effect of F may be involved in the mechanism of Ad secretion, though the secretion of Ad increased with F in the course of labor. The response of fetal adrenal to the stress of labor may be different from that of maternal adrenal since a significant correlation was not noticed between the levels of Ad and cortisol in cord blood as was found in maternal blood. PMID- 2553504 TI - [Effect of thyroid hormone on the beta-receptor of cardiac myocytes isolated from rat ventricle]. AB - INTRODUCTION Most of the symptoms in hyperthyroidism, such as an augmented metabolism, tachyarrhythmias and increased myocardial contractility, resemble those of increased sympathetic beta-adrenergic receptor are a major concern, but the exact mechanisms of the effects remain to be clarified. In the present study, the direct effects of triiodothyronine(T3) on the spontaneous beating and beta receptor in relation to protein synthesis of the cultured ventricular myocytes were investigated. METHODS The cardiac myocytes were prepared from neonatal rat ventricle by enzymatic digestion and incubated at 37 degrees C in the atmosphere of 5% CO2 in air. The culture medium used was DME containing 5% fetal bovine serum and was renewed daily. The spontaneous beating frequencies of the myocytes were measured by the use of optical-video system. The cultured myocytes showed spontaneous beating dependent on Ca2+ concentration about 48 hours after the isolation. Although the beating was irregular and unstable at the beginning, it became regular and steady on the 3rd culture day. The experiments measuring spontaneous beating frequency were performed using the cells of this stage. In these experiments, the effects of T3 on the spontaneous beating frequency were studied after a 15-20 minute incubation with various concentrations of T3 (1 x 10(-10)-1 x 10(-7)M). The spontaneous beating was enhanced by isoproterenol and suppressed by Ca antagonists. The cardiac beta-receptors were identified using the hydrophilic radioligand 3H-CGP-12177 which labels specifically cell surface bound beta-receptors. The effects of T3 on protein synthesis in cultured myocytes were studied by measuring 3H-leucine uptake. RESULTS I. The effects of T3 on the spontaneous beating The effects of T3 on the spontaneous beating were investigated in two different ways. Firstly, the direct effect of T3 was studied by incubating the cells in the medium containing variable concentration of T3. T3 increased the beating frequency dose-dependently, and the maximal accelerating action was observed at the concentration of 10(-8)M. T3 at this concentration increased the spontaneous beating frequency from 131 +/- 25 min to 157 +/- 35 min after the treatment. Higher concentration of T3 induced arrhythmias and seemed to be toxic. Therefore, in the following experiments the concentration of 10(-8)M was used. The cardiac myocytes were incubated in either T3 containing or T3 free medium for 10 days. The incubation medium was renewed every day, and the change in the beating frequency was monitored continuously.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2553505 TI - [Insulin-like growth factor (IGF) receptor in human fetal erythrocytes and fetal rat liver]. AB - In order to clarify the potential role of IGF-I in fetal growth, the dynamics of IGF receptor were investigated in comparison with insulin receptor in human and rat fetuses. In humans, the serum levels of IGF-I measured by IGF-I radioimmunoassay after acid-ethanol extraction were significantly lower in fetuses than in adult controls. Conversely, serum insulin levels in fetuses were not indistinguishable from those in adults. On the other hand, specific binding of 125I-IGF-I to human fetal erythrocytes was 2.3%, which was significantly higher than that of adult women (1.6%). Insulin binding to the erythrocytes was also higher in human fetuses than in adult controls (cord: 3.8%, adult: 3.0%). By Scatchard analysis, the changes in the specific binding of IGF-I and insulin were mainly due to the changes in the binding capacity rather than those in the binding affinity. Additionally, IGF receptor on human fetal erythrocytes was recognized as type I IGF receptor, because the binding of 125I-labelled IGF-I and 125I-labelled IGF-II could be displaced by the addition of cold IGF-I, IGF-II and insulin. In rats, serum levels of IGF-I were also much lower in fetuses than in adult controls. Specific binding of 125I-IGF-I to liver microsomal membranes was 34.3% (per 400 micrograms protein) in fetal rats (D20), which was significantly higher than that of adult rats (3.2%). Scatchard analysis indicated that these changes in 125I-IGF-I binding was chiefly due to the changes in binding capacity. On the other hand, serum insulin levels increased with progress of pregnancy, and specific binding of insulin to microsomal membranes of fetal liver increased on D21 of gestation, mainly due to the increase in binding affinity rather than binding capacity. The bindings of 125I-labelled IGF-I and 125I-labelled IGF-II to fetal liver microsomal membranes were clearly displaced by cold IGF-I and IGF-II but not by excess of cold insulin, indicating that the receptor was type 2 IGF receptor. These results suggest that IGF-I possesses much more receptor in fetal tissues than in adult tissues despite its lower concentration in fetal sera. It is speculated that IGF-I may play an important role in fetal growth and development. PMID- 2553506 TI - [Role of a guanine nucleotide regulatory protein in exocrine pancreatic secretion -effects of cholera toxin and pertussis toxin on cholecystokinin action]. AB - We have recently shown that F- can mimic the actions of cholecystokinin (CCK) on amylase release, Ca2+ mobilization and inositol phosphate generation in pancreatic acinar cells. We have concluded, therefore, that pancreatic CCK receptors may be coupled to the activation of polyphosphoinositide hydrolysis by a guanine nucleotide regulatory protein (N protein), which seems to be sensitive to F-. In the present study, in order to further characterize this N protein coupled to pancreatic CCK receptors, we have examined the effects of bacterial toxins, pertussis toxin (PT) and cholera toxin (CT) on both CCK- and NaF-induced cellular responses in isolated rat pancreatic acini. Neither PT or CT pretreatment of acini affected both CCK- and NaF-stimulated increases in intracellular Ca2+ concentration monitored by quin2. Furthermore, pretreatments of acini with PT and CT didn't alter the effects of CCK on inositol phosphate generation in acini. Similarly, NaF-induced inositol phosphate generation was not changed by these toxin treatments. However, pretreatment procedures employed in this study were considered to catalyze complete ADP-ribosylation of alpha-subunit of the stimulatory (Ns) and inhibitory (Ni) N protein. These results, therefore, strongly suggest that a N protein coupling pancreatic CCK receptors to the breakdown of polyphosphoinositide may be distinct from Ns or Ni like protein. PMID- 2553507 TI - [Clinical studies using a highly sensitive radioimmunoassay for mid-region and carboxy terminus of parathyroid hormone in normal, hypo- and hypercalcemic states]. AB - Parathyroid hormone radioimmunoassay (RIA), specific for mid-region of the PTH molecule, has been proven to be extremely useful for the differential diagnosis of abnormal calcium metabolism. Recently, we developed a highly sensitive RIA for PTH, consisting of PTH antiserum (CH9), 125I labelled Tyr42 hPTH (43-68) and synthetic hPTH (1-84) as standard. This RIA cross-reacted with mid-region and carboxyl terminals of PTH. The within-assay and between-assay coefficients of variation were less than 4.6% and less than 8.6%, respectively. The limit of detection was 50pg/ml. The levels of serum calcium, serum phosphate, serum creatinine, Tmpo4/GFR and creatinine clearance (Ccr) in normal healthy volunteers aged 20 to 50 years remained almost constant and showed 9.24 +/- 0.34mg/dl (mean +/- SD, n = 242), 3.34 +/- 0.38mg/dl (n = 242), 0.870 +/- 0.121mg/dl (n = 242), 3.20 +/- 0.54mg/dl GF (n = 189) and 103 +/- 17ml/min (n = 137), respectively. All healthy volunteers (n = 326) had measurements of PTH in the blood. From 20 to 50 years, normal PTH mean was 374 +/- 97pg/ml (+/- SD, n = 237) and ranged from 180 568pg/ml, and from 60 to 80 years it was 471 +/- 133pg/ml (n = 34) and ranged from 205-737pg/ml. Since we found that PTH was markedly elevated above normal when Ccr was below 40ml/min, and PTH was very significantly correlated with the reciprocal of Ccr (r = 0.8996, P less than 0.001) using a multivariate analysis, all of the patients whose Ccr was higher than 40ml/min were selected and examined in the following studies. Serum PTH values completely separated patients with surgically proven primary hyperparathyroidism (1 degree HPT) from malignant associated hypercalcemia (MAH), and patients with idiopathic hypoparathyroidism (IHP) from pseudohypoparathyroidism (PHP), both of which were diagnosed by Ellsworth-Howard test. PTH values in all of the patients with 1 degree HPT (n = 23) were above normal, but those with MAH (n = 6) were below the normal or lower normal range. PTH values in patients with PHP (n = 7) showed above normal, while those with IHP (n = 5) were below the normal range. PTH was normalized in post operative status in all patients after parathyroidectomy (n = 6). These results indicate that this PTH RIA is extremely useful for the differential diagnosis in diseases with calcium abnormalities. PMID- 2553508 TI - Hydroxylapatite-coated dental implants. Biologic rationale and surgical technique. AB - Implant prosthodontics now offers our patients more benefits than conventional prosthetics. Considerations of bone biology, occlusal principles, occlusal materials, implant design, implant biomaterials, patient health profile, patient bone density and quality, site classification, manufacture quality and ethics, and operator efficiency all go into the prognosis and affect our final product. Emerging biomaterials help the clinician, in certain areas, to achieve a more predictable result. The time-tested principle--such as unloaded healing, atraumatic gentle surgical placement, and machined components--are carried along into the new systems that are being developed for clinical use. The use of the HA coated cylinder has proved to be of extreme value in these past 5 years. It is another step closer to the ideal implant system. PMID- 2553509 TI - [Lowering of the glycemic threshold in pituitary and adrenal response to hyperglycemia in insulinoma]. PMID- 2553510 TI - Pregnancy in a patient with the Klippel-Trenaunay-Weber syndrome: a case report. AB - A rare case of pregnancy in a patient with the syndrome of Klippel-Trenaunay Weber is described. The arterio-venous anomalies in this patient originally occurred in her right leg, but in the course of her first pregnancy she also developed circumscript angiomatosis at the left and right side of the uterus. Her pregnancy was uneventful. However, because of prominent vascular changes in the cervix and lower uterine segment, there appeared to be a cephalo-pelvic discongruency. For this reason a Cesarean section was performed at term. Postoperatively, during heparinization, she had signs of abdominal bleeding, which could be controlled conservatively. There were no signs of intravascular coagulation or cardiac decompensation. A review of the scarce literature on diffuse uterine angiomatosis and angiomatosis occurring as a result of the syndrome of Klippel-Trenaunay-Weber is given. The clinical course and the risks of a pregnancy with this condition are discussed. It is concluded that the angiomatosis based on the syndrome of Klippel-Trenaunay-Weber is less hazardous than a diffuse angiomatosis of the uterus without this disease, the main risk being diffuse intravascular coagulation at or after delivery. PMID- 2553511 TI - Comparison of the effects of magnesium hydroxide and a bulk laxative on lipids, carbohydrates, vitamins A and E, and minerals in geriatric hospital patients in the treatment of constipation. AB - In a crossover study the effects of magnesium hydroxide on serum lipids, carbohydrates, vitamins A and E, uric acid and whole blood minerals were compared with those of a bulk laxative containing plantago rind and sorbitol in 64 constipated, elderly long-stay patients, 55 of whom were receiving diuretics. Hypomagnesaemia occurred in 11 (17%) patients after bulk laxative and in two (2%) patients after magnesium hydroxide treatment. There was a slight reduction in low values of high-density lipoprotein cholesterol and high values of triglycerides after magnesium hydroxide treatment. There were no significant differences in plasma lipids, whole blood minerals or vitamins A and E using either laxative. Negative correlations were found between the increase in serum concentrations of magnesium and glycosylated haemoglobin A1 (P less than 0.02) and the serum level of uric acid (P less than 0.01). These results suggest that the long-term effects of magnesium hydroxide and bulk laxative on the absorption of nutrients may not be significantly different. Magnesium hydroxide, however, may have beneficial effects on lipid disorders, impaired glucose tolerance and hyperuricaemia in magnesium deficiency due to diuretics and thus may be a favourable laxative for use in bedridden geriatric patients receiving diuretics. PMID- 2553512 TI - Transient expression of a Ca2+-activated Cl- current during development of quail sensory neurons. AB - The expression of a calcium-activated chloride current (ICl(Ca)) was studied during the development of the sensory neurons of quail trigeminal ganglia. This current is expressed in 20% of the neurons by the 5th day of embryonic development; it can be found in nearly all neurons by the 7th day and subsequently disappears in half of them. Similar results were obtained with dorsal root ganglion neurons. The disappearance of ICl(Ca) in part of the sensory neurons during development is not due to a selective death of the neurons possessing this current and our results suggest that it is mediated by an interaction of the sensory neurons with their target tissue. PMID- 2553513 TI - Fetal male masculinization in control and prenatally stressed rats. AB - The present experiments were designed to test the hypothesis that males in utero masculinize the development of other males. This effect was examined during fetal development in males from control and prenatally stressed rats. A code identified the number of cervical-flanking males between the target male and the cervical end of the uterus. The male parameters morphology (anogenital distance) and body, adrenal, and testis weights were recorded on the eighteenth and twentieth gestational days and categorized by the cervical-flanking male classification. At Day 18, control fetuses with two cervical-flanking males in utero displayed significantly greater anogenital distance values than did males with no cervical flanking male. At Day 20, control fetuses with two cervical-flanking males had testicular weights significantly greater than those of fetuses with one or zero cervical positioned male. Prenatal stress markedly impaired male fetal development at gestational Days 18 and 20 while suppressing the cervical-flanking male effect. These results confirm and extend previous data that indicate: (a) prenatal stress disrupts normal fetal development, resulting in long-term changes; and (b) androgens via a cervical-flanking blood-flow mechanism influence littermate morphology and sexual development during the prenatal period. Our findings also demonstrate that a general masculinizing effect could not be made across the measured male parameters since the effect of males positioned at the cervical-flanking region in utero appears to be dependent upon maternal sources of variance. Finally, androgens prenatally have an apparent positive interaction with somatic growth. PMID- 2553514 TI - Role of oxygen-derived free radicals in the mechanism of chronic gastric ulceration in the rat: implications for cytoprotection. AB - Oxygen-derived free radicals are cytotoxic and mediate tissue damage. Allopurinol prevents the formation of superoxide radicals and dimethyl sulphoxide (DMSO) scavenges the hydroxyl ones. Intraperitoneal reserpine (5 mg/kg every day for 5 days) produced chronic gastric ulceration in all rats after 4 weeks. Animals gavaged with 1 ml/day of each of 1% allopurinol and 1% DMSO for 4 weeks developed ulceration in 60% of cases; however this ulceration developed in only 20% of animals similarly gavaged with 2% solutions. Rats gavaged with 1 ml/day of each of 5% allopurinol and 5% DMSO for 4 weeks were completely protected against the reserpine-induced chronic gastric ulceration. This protection was not associated with any significant effect on the H+ output of the rat stomach. The results suggest that in the rat, oxygen-derived free radicals are responsible for the development of chronic gastric ulceration and that removing these radicals protects against the said ulceration without influencing acid secretion, i.e. cytoprotection. PMID- 2553515 TI - Mechanism of glucose-induced (Na+, K+)-ATPase inhibition in aortic wall of rabbits. AB - Hyperglycaemia decreases (Na+, K+)-ATPase activity in specific tissues by a mechanism whose effects are prevented by aldose reductase inhibitors and by raising plasma myo-inositol. This mechanism was activated and studied in vitro in normal rabbit aortic intima-media. Raising medium glucose to 10 mmol/l for 60 min inhibited a major component of (Na+, K+)-ATPase-mediated 86Rb+/K+ uptake normally operative in resting aortic intima-media in medium containing normal plasma levels of glucose (5 mmol/l) and myo-inositol (70 mumol/l); 20 or 30 mmol/l glucose had no greater effect. This effect occurred under conditions in which the aortic intima-media's normal myo-inositol content is not detectably decreased. The inhibition was prevented by sorbinil (10 mumol/l) and by raising medium myo inositol from 70 to 500 mumol/l, which had no effect on (Na+, K+)-ATPase activity when the medium glucose remained at 5 mmol/l. Raising medium glucose selectively inhibited a component of (Na+, K+)-ATPase activity that requires medium myo inositol, because it is maintained by a regulatory system through rapid basal phosphatidylinositol turnover in a discrete pool, which is replenished by a fraction of basal de novo phosphatidylinositol synthesis that is selectively dependent on myo-inositol uptake. Medium myo-inositol at a normal plasma level became inadequate to maintain this fraction of basal de novo phosphatidylinositol synthesis [( 1,3-14C]glycerol incorporation) when the medium glucose was raised. When sorbinil was added raising medium glucose did not alter the ability of 70 mumol/l medium myoinositol to maintain the (Na+, K+)-ATPase activity that requires medium myo-inositol.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553516 TI - Transfected rat islet tumour cells express mouse major histocompatibility complex class I antigens functionally. Applicable as "pseudo-syngeneic" targets in the multiple low-dose streptozotocin diabetes model. AB - Rat insulinoma cells clone 5AH-B, were transfected by electroporation with the gene encoding the mouse major histocompatibility complex class I antigen H-2Kb, whereupon stable transfectants were selected and analysed. Data from flow cytometric analyses using three different H-2Kb specific monoclonal antibodies and functional assays using H-2Kb specific alloimmune cytotoxic T cells revealed that the encoded H-2 antigen was expressed in a functional manner. Similar experiments employing the monoclonal antibody OX-18, which recognizes rat major histocompatibility class I molecules, and xenoimmune cytotoxic T cells specific for the endogenously expressed RT1g antigen showed that functional expression of the RT1g antigen was maintained. However, a down-regulation of the expression was observed in H-2Kb positive transfectants, whereas normal expression was retained in Kb negative transfectants. The function of the native promoters of both the endogenous and the transfected class I genes was found to be preserved in the transfectants as assessed by the response to stimulation with interferon-tau. The present study was unable to confirm the reports of RIN specific lysis by T cells from multiple low dose streptozotocin diabetic mice. Even in the presence of the syngeneic restriction element no lysis was observed. We conclude, that rat insulinoma cells clone 5AH-B, are able to integrate a foreign class I antigen gene and express the encoded product functionally. The data also suggest the possibility of creating major histocompatibility antigen positive rat insulinoma cells which are RT1g negative. Such transfectants will be of great potential value for the dissection of cell mediated B cell destructive processes. PMID- 2553517 TI - Ca++ antagonists distinguish different requirements for cAMP-mediated gene expression in the cellular slime mold, Dictyostelium discoideum. AB - Cyclic AMP is essential for the accumulation of many prespore mRNAs and can advance the time of appearance of mRNAs specifically enriched in prestalk cells. Additionally, when late-developing cells are washed free of cAMP, a number of growth phase mRNAs reaccumulate. This reaccumulation can be suppressed by cAMP. These effects of cAMP are all mediated through the cell surface cAMP receptor and can occur under conditions where the receptor-associated adenylate cyclase is inactive, indicating that the initial intracellular transduction event necessary for expression of these mRNAs does not depend upon cAMP synthesis. The dihydropyridine derivatives, nifedipine and nitrendipine, are highly specific Ca++ channel blockers. They are shown here to prevent the influx of Ca++ from the external medium that occurs in response to cAMP binding to the cell surface receptor during development. These two compounds as well as another Ca++ antagonist, 8-N,N-diethylamino)octyl-3,4,5-trimethoxy-benzoate (TMB-8) and a calmodulin inhibitor, N-(6-amino-hexyl)-5-chloro-1-naphthalene sulfonamide (W7), all specifically decrease cAMP-mediated prespore mRNA accumulation in a dose dependent manner. They also prevent cAMP from suppressing the expression of the growth phase genes. The growth phase mRNAs reaccumulate in cAMP-treated cells in the presence of increasing concentrations of these drugs. By contrast, cAMP induction of the pre-stalk-enriched mRNA is not as significantly affected by these agents. These results raise the possibility that the cell surface cAMP receptor can couple to different signal transduction systems and thereby induce or suppress the expression of different sets of cAMP-regulated genes during development. PMID- 2553518 TI - Conditions that elevate intracellular cyclic AMP levels promote spore formation in Dictyostelium. AB - We have been using sporogenous mutants of Dictyostelium discoideum strain V12M2 to study regulation of cell fate during terminal differentiation of spores and stalk cells. Analyses of intracellular cAMP accumulation, cAMP secretion, cAMP binding to cell surface receptors, and chemotactic sensitivity to exogenous cAMP during aggregation showed that all of these functions were identical in V12M2 and HB200, a sporogenous mutant. We used several methods of altering intracellular cAMP levels in HB200 cells to test the hypothesis that intracellular cAMP levels affect cell fate. First, HB200 amoebae were treated with 5 mM caffeine for 4 h during growth, washed, and allowed to develop in the absence of caffeine. Treated cells had normal levels of intracellular cAMP and adenylate cyclase activities at the beginning of differentiation; by 6 h development, they contained two to three times more intracellular cAMP and two times more GTP-dependent adenylate cyclase activity than untreated cells. However, their level of basal Mn++-dependent adenylate cyclase activity was the same as untreated controls. Thus, treatment of growing HB200 amoebae with caffeine for only 4 h leads to hyperinduction of a GTP dependent regulator (or inhibition of a negative regulator) of adenylate cyclase during subsequent differentiation, without induction of basal activity. The fraction of amoebae forming spores increased twofold when HB200 amoebae were treated with caffeine during growth. Spore (but not stalk cell) differentiation by such treated cells was blocked by inhibitors of cAMP accumulation. Second, cells grown on nutrient agar accumulated higher levels of intracellular cAMP and formed more spores in vitro than cells grown in shaken suspension.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553519 TI - Distribution and developmental transition of phosphoglycerate mutase and creatine phosphokinase isozymes in rat muscles of different fiber-type composition. AB - Phosphoglycerate mutase and creatine phosphokinase have in mammals three isozymes (types MM, MB and BB) with similar tissue distribution and developmental transition in muscle cells. To assess whether the phenotype and the developmental switch of these isozymes differ in the diverse types of muscle fibers, the enzymatic activities and the isozyme patterns, analyzed by cellulose acetate electrophoresis, have been determined in rat soleus, extensor digitorum longus and gastrocnemius muscles during postnatal development. Both phosphoglycerate mutase and creatine phosphokinase activity increased in the three muscles, the increase in extensor digitorum longus and gastrocnemius being higher than in soleus. For the two enzymes the increase in activity was due to the progressive increment of the muscle-specific forms. It is concluded that whereas phosphoglycerate mutase and creatine phosphokinase type-B subunits are present at similar levels in both type I and type II muscle fibers, phosphoglycerate mutase and creatine phosphokinase type-M subunits exhibit much higher levels in type II fibers. PMID- 2553520 TI - [Effects of hypoxia on the production of angiotensin converting enzyme in decompensated liver cirrhosis]. AB - The high angiotensin-converting activity, in 13 of the 17 patients afflicted with hepatic cirrhosis, has been put in relation to the hypoxia influence. The insufficient cession of O2 seems to stimulate the enzyme activity, especially for the patients who simultaneously present low values of blood pseudo cholinesterase. The AA think that this behaviour is to be ascribed to a deficiency of the hepatic proteic synthesis of vasoactive substances, such as the angiotensins. PMID- 2553521 TI - Neuromuscular diseases of AIDS. AB - Neuromuscular diseases are common in acquired immune deficiency syndrome (AIDS). Although the clinical incidence of peripheral neuropathy has not been systematically studied, various reports suggest that up to 40% of AIDS patients have clinical symptoms. Biopsy and autopsy studies have shown an inflammatory neuropathy with a variable component of demyelination and axonal loss. Evidence of direct involvement by the human immunodeficiency virus (HIV) is scant. Immunosuppression followed by cytomegalovirus (CMV) infection appears to be a direct cause of polyradiculoneuropathy and perhaps other forms of peripheral neuropathy in AIDS. The clinical incidence of myopathy in AIDS is less clear, and clinically less appreciated than the neuropathy. Scattered reports have identified an inflammatory myopathy that does not appear to be due to direct HIV infection, but could be mediated by another human retrovirus. HIV seropositive patients being treated with antiviral drugs develop a unique set of neuromuscular diseases that must be distinguished from the non-drug-related conditions. PMID- 2553522 TI - Role of peptidergic neurons in ocular herpes simplex infection in mice. AB - Peptide-containing nerve fibers (peptidergic fibers) abundantly innervate the mammalian cornea. We investigated their role in ocular herpes simplex infection in mice by using capsaicin, which causes degeneration and permanent loss of peptidergic neurons in neonates and temporary peptide depletion in adult animals. The corneas of neonatally denervated BALB/c mice were observed for capsaicin induced keratitis at 11-14 wk of age and were then infected bilaterally with herpes simplex virus 1 (HSV-1); trigeminal (TG) ganglia were cocultivated 6 wk later to establish the rate of latent infection. We also applied capsaicin eye drops to adult BALB/c mice that had been infected with HSV-1 6 wk earlier, and measured viral shedding before, and 3 days and 2 months after, administration of capsaicin drops; TG ganglia of these animals were cocultivated at 3 days and 2 months after capsaicin application. Neurotrophic keratitis was found in 81% of neonatally denervated animals; mortality rate due to HSV-1 infection was reduced from 80% in the controls to 24% in the capsaicin-treated group, and recovery of latent virus by cocultivation was reduced by 50%. Viral shedding could not be produced by capsaicin eye drops in adult animals infected with HSV-1. However, recovery of latent virus was significantly reduced in TG ganglia sampled 3 days and 2 months after capsaicin drops were instilled. Our findings suggest 1) that peptidergic fibers play a crucial role in the establishment of trigeminal HSV-1 latency and 2) that reactivation of latently infected ganglia can be inhibited by topical capsaicin. PMID- 2553523 TI - Scirrhous carcinoma of the stomach: a clinical and pathological study of 106 surgical cases. AB - Clinical and pathological characteristics of scirrhous carcinoma of the stomach were studied in 106 cases treated by gastrectomy between 1973 and 1983. The male to female ratio was 0.58. The percentage of scirrhous carcinomas to all gastric carcinomas resected in the same period was three times higher in females than males. The age distribution of the patients suggested that there were two peaks in the forties and sixties in the male, and in the thirties and fifties in the female. The incidence of scirrhous carcinoma in all types of gastric carcinoma was significantly higher in the twenties, thirties and forties compared to the lowest incidence in the seventies. In the female group the primary lesion had a tendency to be adjacent to the fundic gland area and to avoid intestinal metaplasia. In the male the opposite was recognized. Cancer nests with single cells or only several cells were common in this type of carcinoma. These findings suggest that there might be two biologically different scirrhous carcinomas both in the male and the female, the appearance of single carcinoma cells might be favored by female sex hormones and young ages, and not only the original gastric mucosa but also mucosa with intestinal metaplasia could be precursors of single carcinoma cells. PMID- 2553524 TI - Role of superoxide and hydroxyl radicals in rat gastric mucosal injury induced by ethanol. AB - It has been reported that oxygen-derived free radicals play an important role in the pathogenesis of mucosal injury in the small intestine as well as in the stomach. The aims of this study were to test whether ethanol-induced damage in the rat stomach was prevented by the administration of (1) superoxide dismutase (SOD; a scavenger of superoxide radicals), (2) allopurionol (ALP; an inhibitor of xanthine oxidase), (3) dimethyl sulfoxide (DMSO; a scavenger of hydroxyl radicals). SOD significantly decreased the ulcer index from 100 +/- 8.5% (control) to 39.6 +/- 8.2% (P less than 0.001). Ethanol-induced damage was reduced by the administration of ALP by 37.4% (P less than 0.01). DMSO also diminished the ulcer index from 100 +/- 8.5% (control) to 31.6 +/- 5.8% (P less than 0.01). Histochemical studies supported these results. A scanning EM study, however, revealed that surface epithelial cells were not protected by SOD against ethanol-induced damage. These results demonstrated that SOD, ALP and DMSO had the ability to protect gastric mucosa against ethanol-induced injury. Accordingly, oxygen-derived free radicals may be involved in the pathogenesis of ethanol induced gastric mucosal damage. Surface epithelial cells, however, were not protected even by SOD against ethanol-induced injury. PMID- 2553525 TI - Implications of hyperechoic lesions in small hepatocellular carcinoma. AB - Of 34 solitary small hepatocellular carcinomas (HCC) 2 cm in diameter or less, 13 with hyperechoic lesions were observed serially by sonography, and 11 of these were examined histologically. Serial examination showed that hypoechoic areas appeared at the periphery of or within, the hyperechoic tumor, and that these areas expanded more with tumor growth than the hyperechoic areas as if compressing or displacing the existing hyperechoic areas. Histologically, the hyperechoic lesions were composed mostly of well-differentiated cancer cells containing fat droplets, whereas the hypoechoic lesions were composed of cancer cells without fat droplets. In the two tumors that were formed almost completely of cancer cells showing fatty metamorphosis, cancer cells without fat droplets proliferated mainly in the periphery of the tumor. These findings suggest that, in hyperechoic HCC, cancer cells with fat droplets appear in the early stage of HCC, and probably change into concer cells without fat droplets by the time that a certain tumor size is reached, with gradual displacement by the latter type of cell during tumor growth. PMID- 2553526 TI - Effect of arachidonate derivatives on cell viability in isolated rat hepatocytes: cytotoxicity of 15-hydroperoxyeicosatetraenoic acid. PMID- 2553527 TI - Glycogen-enzymes containing bodies in type 2 fibres of tenotomized muscles in the rat. AB - Inclusion bodies containing glycogen-enzymes were found in 30 to 60% of type 2 fibres of tenotomized calf muscles (m. gastrocnemius, m. soleus, m. plantaris) in rats, using histochemical reactions. The bodies appeared within 1 week after the tenotomy and were localized both in the central and the subsarcolemmal regions and rarely extruded into the extracellular space. These aggregates are 3 to 15 microns in length and 2 to 11 microns in diameter. In addition to glycogen, these bodies also contained various enzymes of the glycogen metabolism such as phosphorylase, a branching enzyme, and glucose-6-phosphatase, but showed no NADH reductase, lactate dehydrogenase, or myofibrillar ATP-ase activity. The results indicate that glycogen-enzymes containing bodies are a degenerative phenomenon, which occurs only in type 2 fibres of the tenotomized muscles. PMID- 2553528 TI - [Prevention of thrombosis in gynecology: double-blind comparison of low molecular weight heparin and unfractionated heparin]. AB - The safety and efficacy of low molecular heparin (Sandoz AG, Nurnberg), given 1500 aPTT once daily (and two placebo injections), in preventing postoperative venous thromboembolism, was assessed against sodium heparin at a dose of 5000 IU three times daily, in a unicenter double-blind randomized study. 300 patients, scheduled for major gynecological surgery, were included in this study. Two patients (1.3%) of 150 patients developed deep vein thrombosis in the LMWH-group, the corresponding figure for sodium heparin was 6 patients (4.0%). There was no statistically significant difference between the two groups in respect to the bleeding variables such as postoperative drainage, blood transfusions and haematoma. The antithrombotic effects were assessed with the anti-Xa assay and LMWH-fractions have shown a high antifactor Xa/activated partial thromboplastin time (aPTT) specific activity (2:1) compared to unfractionated heparin (1:1). It is concluded that one single daily injection of LMWH (without dihydergot) provides a convenient safe and effective prophylaxis against thromboembolism in gynecological surgery. PMID- 2553529 TI - The Drosophila zeste locus is nonessential. AB - Diepoxybutane-induced mutations of the Drosophila zeste locus were generated in an effort to obtain a null allele. Of 33 mutations of this X-linked gene isolated, 16 were associated with multilocus deletions of zeste and adjacent complementation groups, while the remainder were defects restricted to zeste undetectable by Southern blot analysis. Two of these multilocus deletions (Df(1)zdeb3 and Df(1)zdeb92) were employed in the synthesis of females completely deleted for zeste. Such "zesteless" flies were produced, though at frequencies lower than Mendelian expectations. zeste-deleted females are fertile, and can give rise to zeste-deleted female progeny. These results demonstrate that the product of the zeste gene is not essential to viability or to female fertility, even if absent both as a maternal contribution and as a product of the zygotic genome. However, the possibility that zeste may influence relative viability cannot be excluded. In spite of previous in vitro indications that the zeste protein may activate transcription of the Ultrabithorax (Ubx) gene, zeste deleted flies are Ubx+ in phenotype. This suggests that the zeste protein normally is either a very weak transcription factor, or that its function can be substituted by that of other regulatory proteins. PMID- 2553530 TI - Limbic encephalitis. A rare presentation of the small-cell lung carcinoma. AB - Two patients with an acute organic brain syndrome and accompanying neurological symptoms are described. Extensive work up showed that both patients suffered from small-cell lung cancer. Cerebral metastases were absent. Following chemotherapy and radiotherapy to the primary tumor one of the two patients showed a complete remission of psychiatric symptoms for one year. A paraneoplastic origin of this syndrome, in the literature known as limbic encephalitis, is postulated. The exact cause of this syndrome is yet unknown. Recent research reveals data indicating an immunological pathogenesis. The major clinical importance of this (neuro)-psychiatric syndrome is that its appearance may serve as a warning sign for an occult malignancy; furthermore, effective treatment of the primary malignancy can reverse the encephalitis. Thus antitumor therapy can result in a prolonged survival and considerably improved quality of life. PMID- 2553531 TI - [Genetic instability of determinants of resistance to chloramphenicol and kanamycin in Streptomyces lividans 66]. AB - The frequency of chloramphenicol-sensitive variants (Cmls) in Streptomyces lividans 66 is very high (0.57%). Correlation between chloramphenicol sensitivity and deamplification of PstI fragment with the length of 4.82 kb (RES1 genetic element) was shown. However, in some Cmls variants there was no RES1 deamplification. It was noted that in the cells of the Cmls variants isolated the levels of kanamycin and neomycin resistance determined by the Kanr determinant in the pSU17 plasmid were different. Expression of Kanr and Neor determinants inserted via pSU17 plasmid into the cells of Cmls variants was studied and three classes of chloramphenicol-sensitive variants were defined. After transformation of pSU17 plasmid into cells of Cmls variants of the class I, expression of Kanr and Neor genes, similar to that in S. lividans 66, was observed. The resistance level in Cmls variants of the class II was intermediate. In the cells of the class III no expression was noted. Cmls strains of classes I and II were unstable and those of the class III with impaired expression of Kanr and Neor genes were formed with high frequency. Cmlr variants formed from Cmls strain of the class III were studied. Two types of Cmlr variants were detected. Variants of the first type were identical to S. lividans 66 by their properties. The frequency of Cmls variants occurring in the cells of the first type was similar to that in S. lividans 66. The second type included pseudo-revertants. They were unstable and generated amplifications of the 5.7 kb fragment with high frequency. PMID- 2553532 TI - [Intracellular distribution of nucleotide sequences, homologous to mobile dispersed genes]. AB - Full-size hybrid RNA-DNA molecules of mdg1, mdg3 and copia have been discovered in cultured cells of Drosophila. The intracellular distribution of these molecules is considered. Hybrid molecules are found both in the nuclei and cytoplasm, the latter comprising the major portion of molecules. The membrane subcellular fraction enriched by poly(A) RNA and poly(A) RNA-DNA molecules of retretransposons is isolated from cytoplasm. The full-size linear double-stranded DNA of mdg1 is found in the same fraction. The data obtained make it possible to suppose that the process of reverse transcription on the mdg RNA is the way of regulation of mdg expression in the cell. PMID- 2553533 TI - [Participation of the uvrD gene in the precise excision of the Tn9 transposon]. AB - HfeTn5-(04,06) and IfeTn9 mutations increase efficiency of precise excision of Tn5, Tn10 and decrease that of Tn9. These mutations have been mapped in uvrD gene. In LFETn9 and UVRE502 mutants, the multicopy plasmid pEM61 carrying the cloned uvrD gene complements LFETn9- phenotype (Low Frequency Excision of Tn9). These results indicate that the uvrD gene product plays different role in excision of transposons with long and short inverted repeats. The mechanism of this effect is discussed. PMID- 2553534 TI - Comparison of the regulation of P elements in M and M' strains of Drosophila melanogaster. AB - M and M' strains of Drosophila melanogaster in the P-M system of hybrid dysgenesis were compared in two series of tests, with the following results. (1) The singed-weak hypermutability regulation test showed that M' strains had lower P excision rates than M strains, suggesting that P-elements repression must occur in M' strains although it is not detectable by gonadal dysgenesis assays. (2) The evolution of mixed P+M and mixed P+M' populations was compared, using a strong P strain. The P+M cultures invariably evolved in a few generations into strong P cultures, while the P+M' cultures evolved into P-type cultures with reduced P factor potentials. However, after 30 generations of culture, both these types of mixed cultures had similar P copy numbers, suggesting that regulation of copy number had occurred in them. PMID- 2553535 TI - Physical and biological consequences of interactions between integration host factor (IHF) and coliphage lambda late p'R promoter and its mutants. AB - The integration host factor (IHF) binds to a site (ihf) that overlaps the -35 region of the phage lambda late rightward promoter (p'R). This interaction represses p'R-promoted transcription, both in vivo and in vitro. In vivo repression was observed when a plasmid carrying both p'R and the galK reporter gene was transfected into IHF+ or IHF- hosts. In vitro repression of transcription by IHF was observed only with linear, but not with supercoiled wild type p'R templates. When binding to ihf, IHF imposes a strong bend on the DNA and protects this site from cleavage by neocarzinostatin, pancreatic DNase I, and hydroxyl radicals, as assessed by footprinting experiments. Both the functional and nonfunctional p'R mutants, in which the upstream part of the -35 region was replaced by an EcoRI linker, show modified behavior toward IHF. Some are more sensitive to IHF-mediated repression, even in the supercoiled form, while others have lost their affinity for IHF. We conclude that IHF binding depends not only on the consensus ihf sequence, but also on a suitable combination of the sequences of both ihf and neighboring regions, together with the DNA conformation, which includes both natural and imposed bends in DNA and the degree of supercoiling. Based on most of the present data, it is difficult to predict the relationship between the ihf sequence and IHF interaction, since two very different sequences (less than 50% homology) show strong IHF binding, whereas very similar sequences (80-87% homology) show a very different behavior. However, the hydroxylradical footprinting data show that three A + T-rich sequences are protected by IHF: the central sequence, which overlaps the -35 region of p'R, and two flanking sequences removed by one helix turn. All three sequences are located on the same face of the helix, and the amino acid side chains of IHF seem to occupy the narrow minor groove. A novel consensus sequence is proposed. PMID- 2553536 TI - Hepatitis B virus surface antigen as a reporter of promoter activity. AB - The coding sequence for the hepatitis B virus surface antigen (HBsAg) was used as a new reporter gene for studies on eukaryotic promoter activity and upstream regulatory sequences. The data observed in transfection assays were comparable to results obtained with conventional chloramphenicol acetyltransferase (CAT) assays, as was demonstrated using various transcriptional regulation sequences. The expression of HBsAg as a reporter protein offered some advantages: (i) In transient expression assays, a time course of promoter activity depending on variable culture conditions could be monitored over a period of time, since the HBsAg was secreted into the culture supernatant. (ii) Evaluation of HBsAg from supernatant aliquots and quantification of the corresponding promoter activities could be performed easily, using the very sensitive and readily available diagnostic HBsAg kits. (iii) In contrast to the conventional CAT assay, the cells remained available for further tests, e.g., Western blot, immunofluorescence or transcript analysis. Characteristics of several Epstein-Barr virus (EBV) promoters, depending on the virus state of EBV-positive B-cells (latency, chemical induction, lytic superinfection, trans-activation), were assayed using the HBsAg reporter system. PMID- 2553537 TI - Neutral selection of transfected mammalian cells using tissue plasminogen activator gene expression. AB - The gene that produces the proteolytic enzyme, tissue plasminogen activator (tPA), was used as a selectable marker for transfection. Cells that were successfully transfected with plasmids containing a gene for tPA (tpa) produce plaques in the caseinolysis assay. Two tpa-containing plasmids were constructed and used to transfect a variety of cell types. One plasmid contained the promoter region of simian virus 40 and the other contained the Moloney murine leukemia virus promoter. No significant difference in transfection frequencies was found for the two plasmids. However, 80% of the cell types tested produced plaques. This system allows for the identification and isolation of transfected cells under conditions that are not lethal to nontransfected cells. PMID- 2553538 TI - Respiratory-competent yeast mitochondrial DNAs generated by deleting intergenic regions. AB - Two respiratory-competent yeast strains having mitochondrial (mt) DNA characterized by single non-overlapping deletions, encompassing intergenic sequences, have been crossed. Diploid daughter clones have been screened by electrophoresis of mtDNA fragments, and a respiratory-competent clone (ER8.75), having a recombinant small mtDNA with both parental deletions, has been detected. ER8.75 mtDNA lacks around 20% of wild-type intergenic sequences, encompassing three ori/rep sequences. This mutant could be helpful in analyzing the organelle genome and, particularly, the function of intergenic sequences. PMID- 2553539 TI - Interplay of novobiocin-resistant and -sensitive DNA gyrase activities in self protection of the novobiocin producer, Streptomyces sphaeroides. AB - The novobiocin (Nb)-producing organism, Streptomyces sphaeroides, possesses two gyrB genes: gyrBS and gyrBR (encoding the DNA gyrase B subunit-the normal target for Nb) whose products differ in their response to the drug. Novobiocin-sensitive gyrase is the predominant form of the enzyme in this strain and is produced constitutively but at variable levels, whereas Nb-resistant gyrase appears when growth takes place in the presence of the drug. The promoter isolated from the Nb resistance determinant responds sharply to changes in DNA topology, being activated when the (negative) superhelical density is reduced and vice versa when the supercoiling of DNA is increased. Thus, resistance to Nb in S. sphaeroides is induced by a reduction in DNA supercoiling due to the action of autogenous drug on the sensitive gyrase. PMID- 2553540 TI - Transposon mutagenesis of baculoviruses: analysis of TFP3 lepidopteran transposon insertions at the FP locus of nuclear polyhedrosis viruses. AB - We report the complete sequences of two representatives of the TFP3 transposable element family of the lepidopteran, Trichoplusia ni. These elements were isolated as insertions mobilized from the Lepidopteran host genome into two closely related nuclear polyhedrosis viruses (NPV) during infection. Both elements inserted within the 500-bp FP locus of the respective viral genomes (map units 36.0 to 37.0), causing a distinctive plaque morphology phenotype and the loss of a 25-kDa viral-specific protein. Both insertions occurred at the identical TTAA target site in the respective genomes, in the same relative orientation, and are flanked by 15-bp imperfect inverted repeats. The inserted elements interrupt the 25K open reading frame (ORF). One of these FP mutants undergoes spontaneous reversion. Sequence analysis at the excision site of a spontaneous revertant demonstrates that the TFP3 elements are capable of precise excision, restoring the expression of the 25-kDa protein. We also compare the sequences of the 25K genes of the Autographa californica and Galleria mellonella viruses (AcMNPV and GmMNPV, respectively). The 25K gene sequences diverge in five areas, resulting in an additional EcoRV and TaqI site within the GmMNPV 25K gene, and extension of the ORF for an additional 2 amino acids at the C-terminus of the predicted GmMNPV 25 kDa protein. The phenomenon of transposon mutagenesis of Baculovirus genomes provides a unique opportunity for analysis of transposon mobility. PMID- 2553541 TI - Multiple high-frequency transpositions of Tn5 in Serratia marcescens 274 from a thermosensitive replication mutant of plasmid R388. AB - We have used the broad-host-range conjugative suicide plasmid vector described by Sasakawa and Yoshikawa [Gene 56 (1987) 283-288] for transposon mutagenesis of Serratia marcescens 274. We report multiple transposition events of Tn5 from this vector. In addition, the unusual pattern of Tn5 transposition might provide an insight into its regulation. PMID- 2553542 TI - Nucleotide sequence analysis of IS256 from the Staphylococcus aureus gentamicin tobramycin-kanamycin-resistance transposon Tn4001. AB - Resistance to the aminoglycosides gentamicin, tobramycin and kanamycin (GmTmKmR) in Australian clinical strains of Staphylococcus aureus is commonly carried on the composite transposon Tn4001. The resistance gene aacA-aphD of Tn4001, which encodes a bifunctional AAC(6')-APH(2") modifying enzyme, is flanked by two 1324 bp inverted repeats, IS256L and IS256R, that are identical in sequence. Analysis of the IS256 sequence revealed structural features characteristic of IS elements including 26-bp imperfect terminal inverted repeats and a single open reading frame with coding capacity for a 45.6 kDa protein. The nucleotide sequence of IS256 described here, together with the sequence of the aacA-aphD gene reported previously [Rouch et al., J. Gen. Microbiol. 133 (1987) 3039-3052], completes the entire sequence of Tn4001, which totals 4566 bp. PMID- 2553543 TI - Antioxidant capacity of diethyldithiocarbamate in a metal independent lipid peroxidative process. AB - 2,2'-Azobis-[2-amidinopropane] initiated lipid peroxidation of egg yolk phosphatydil choline liposomes was measured by oxygen uptake and the emitted visible luminescence. Lipid peroxidation involved a chain process (kinetic chain length = 49 +/- 11) and its rate was independent of added Fe ions. Diethyldithiocarbamate (DDC) behaved as an efficient inhibitor in the microM range, being able to trap 1.05 +/- 0.25 free radicals per added molecule. The efficiency of DDC was also independent of Fe addition to the system. These results indicate that DDC is able to trap the chain carrying free radicals, showing that this compound, besides being a powerful metal chelator, is also an efficient free radical scavenger. It is proposed that the relevant step in this process involves an electron transfer from DDC to the peroxy radical LOO., LOO. + DDC- ----LOO- + DDC. followed by protonation of LOO- and dimerization of the DDC. radical. PMID- 2553544 TI - Electron spin resonance detection of oxygen-centred radicals in murine macrophages stimulated with bacterial endotoxin. AB - The production of oxygen radicals by Bacille-Calmette-Guerin primed mouse macrophages stimulated with bacterial endotoxin has been investigated. Superoxide radicals were spin-trapped in this system with dimethylpyrroline-N-oxide after a lag period of 20-40 minutes. The electron spin resonance signals due to the superoxide radical adduct could be inhibited by superoxide dismutase but not by catalase. PMID- 2553545 TI - Do humans neutrophils form hydroxyl radical? PMID- 2553546 TI - Requirement for lipopolysaccharide for enhanced in vitro superoxide producing competence in macrophages from normal and malaria (Plasmodium chabaudi) infected mouse spleen. AB - Nucleated cells from mouse spleens were suspended in RPMI-1640 medium and plated in polystyrene tissue culture dishes with or without lipoplysaccharide (LPS, endotoxin) and incubated at 37 degrees C for 3 h in a humid incubator under 5% CO2. Adherent spleen cells obtained from uninfected control or Plasmodium chabaudi-infected mice showed negligible superoxide (O2) production in vitro, in response to triggering with phorbol myristate acetate (PMA). However, O2 release increased in a concentration-dependent fashion as LPS was introduced above a threshold of about 10 ng/ml. This LPS dependence is at least 100 times more than that required by mouse peritoneal macrophages and 10(6) times greater than the requirement reported for human monocytes. These findings emphasize the differences among the three mononuclear cells, and also suggest that LPS could prove useful in the isolation and culture of other metabolically competent cells. PMID- 2553547 TI - In vitro scavenger effect of dihydroquinoline type derivatives in different free radical generating systems. AB - The non-toxic and water soluble dihydroquinoline type antioxidants: CH 402 (Na 2,2-dimethyl-1,2-dihydroquinoline-4-yl methane sulphonate) and MTDQ-DA (6,6 methylene bis 2,2-dimethyl-4-methane sulphonic acid: Na-1,2-dihydroquinoline) were studied in various in vitro tests in which oxygen free radicals were generated. Both compounds were shown to scavenge superoxide radical anions O2- produced in aqueous solution by pulse radiolysis with rate constants k (O2- + MTDQ-DA) = 4.10(8) dm3 mol-1 s-1 and k (O2- + CH 402) = 1.5.10(7) dm3 mol-1 s-1. CH 402 and MTDQ-DA reduced the H2O2 produced in the glucose-glucose oxidase reaction, which was detected by the luminol + hemin reaction with a chemiluminometric method. The dihydroquinoline type substrates inhibited the NADPH-induced and Fe3+ - stimulated lipid peroxidation and the ascorbic acid induced non-enzymatic peroxidation pathways in microsomal fractions of rat and mouse liver. PMID- 2553548 TI - The mechanism of cytochrome C reduction by alkyl radicals. Evidence for multiple reaction pathways. AB - The reactions of the hydroxyalkyl radicals .CH2OH and (CH3)2.COH with oxidized cytochrome c are for more complex than previously reported. Analysis of the pulse radiolytic data by kinetic modelling revealed that only about 40% of the alkyl radicals reduce the ferric iron chromophore. Altogether, four different reactions have to be considered for the disappearance of the alkyl radicals, only two of which affect the metal site. The data show that these radicals, similar to the much more reactive hydrated electrons and hydrogen atoms, are capable to react with biological macromolecules in diverse ways. PMID- 2553549 TI - Hydrogen atom formation by ultrasound in D2O solutions of nitrone spin traps. AB - To elucidate the mechanism of ultrasonically induced H atom formation in D2O solutions of nitrone spin traps, alpha-phenyl-N-tert-butylnitrone (PBN) and alpha (4-pyridyl-1-oxide)- N-tert-butylnitrone (POBN) were exposed to 50 kHz ultrasound in the presence of Ar, and the ratio of the ESR signal of the H-spin adduct to that of the D-spin adduct (H/D ratio) was examined. The H/D ratio increased with the concentration of the spin traps. The magnitude of the H/D ratios correlates with the hydrophobicity of the spin traps. The H/D ratios for the more hydrophobic spin trap, PBN, are larger than those for POBN at all concentrations. Xenon, which has a lower thermal conductivity than argon, was employed for creating higher final temperatures of the cavitation bubbles. For the less hydrophobic spin trap POBN the H/D ratio is lower for xenon than for argon. A similar result was found for PBN at lower concentrations. These results show that the H adducts of PBN and POBN are formed from the spin traps or their decomposition products by homolytic scission of C-H bonds due to pyrolysis. PMID- 2553550 TI - Direct detection of peroxyl radicals formed in the reactions of metmyoglobin and methaemoglobin with t-butyl hydroperoxide. AB - The reaction of metmyoglobin and methaemoglobin with t-butyl hydroperoxide has been investigated using stopped-flow e.s.r. spectroscopy. The major species observed immediately after mixing has been identified as the basis of its g value (2.014) and line width as being due to a peroxyl radical. The detection of this species under anaerobic conditions suggests that this species is the t-butyl peroxyl radical rather than a secondary species. The immediate observation of this species on mixing suggests that this species is arising via a reaction involving the intact haem moiety rather than via 'free iron' produced via oxidative damage. Detection of a second, oxygen-dependent, species, which is formed at a slower rate, in the metmyoglobin reaction suggests that the initial peroxyl radical subsequently reacts with the protein to give a protein-derived radical. PMID- 2553551 TI - Free radical scavenging by myocardial fatty acid binding protein. AB - Recent investigations have indicated the presence of a fatty acid binding protein (FABP) in mammalian heart. This protein binds free fatty acids and their esters with high affinity, however, its physiological role remains unknown. Since FABP constitutes a significant amount of cystolic protein, it is likely that it would be a target for free radical attack. To test this hypothesis, FABP was examined for scavenging against free radicals such as the superoxide anion (O2-), hydroxyl radical (OH.) and hypochlorite radical (OCl.) which may be present in an ischemic reperfused heart. Our results suggest that FABP scavenges O2-, OH. and OCl. as indicated by the FABP inhibition of O2- -dependent reduction of cytochrome c, OH. dependent hydroxybenzoic acid formation and OCl.-mediated chemiluminescence response. FABP was found to be a more potent scavenger of these free radicals compared to bovine serum albumin. Furthermore, FABP was more effective in scavenging OH. than O2-, and inhibited OH. mediated lipid peroxidation process. These results indicate that FABP can scavenge free radicals which may be present in an ischemic/reperfused heart and, thus, may play a significant physiological role in the heart during ischemia and reperfusion. PMID- 2553552 TI - Application of the electrochemistry of cytochrome c to the measurement of superoxide radical production. AB - An electrochemical sensor for the measurement of the superoxide anion produced as a result of the respiratory burst of neutrophils is described. The reduction of cytochrome c by the superoxide ion was monitored using a surface-modified gold electrode. The current produced was shown to be superoxide-specific and proportional to the number of neutrophils in both purified neutrophil preparations and plasma. PMID- 2553553 TI - Prostaglandin E2 and leukotriene B4 synthesis by peripheral leucocytes in alcoholics. AB - Alcohol inhibits phospholipase (PL) activity in a number of animal models. We have therefore measured prostaglandin E2 (PGE2) and leukotriene B4 (LTB4), liberated by stimulated peripheral blood mononuclear cells (PBMC) and neutrophils respectively in chronic alcoholics and in control subjects. Peripheral blood mononuclear cells from alcoholics produced less PGE2 (p less than 0.01) and neutrophils produced less LTB4 (p less than 0.025). Reduced PGE2 production by PBMC of alcoholics was corrected by the addition of exogenous arachidonic acid (p less than 0.005) whilst neutrophil LTB4 production remained lower in the alcoholics (p less than 0.01). Percutaneous liver biopsies were undertaken in the 20 alcoholics having abnormal liver function tests. Prostaglandin E2 biosynthesis was lower in PBMC from patients with alcoholic hepatitis than with alcoholic cirrhosis (p less than 0.05). Analysis of PBMC fatty acid composition demonstrated that endogenous arachidonate and linoleate contents were not significantly different in alcoholics and controls. Cells from controls and alcoholics were incubated with 0, 50 and 150 mmol/l ethanol for two hours but there was no alteration in PGE2 or LTB4 biosynthesis. In summary, we found reduced eicosanoid production by peripheral leucocytes in alcoholics, supporting the hypothesis that chronic alcohol consumption either inhibits membrane bound phospholipase activity or enhances, alternatively, catabolism of eicosanoids. This phenomenon is more marked in alcoholic patients with hepatitis than in those with cirrhosis alone. PMID- 2553554 TI - Changes in serum calcium, magnesium, cyclic AMP and monoamine oxidase levels during pregnancy and under prolonged ritodrine treatment for preterm labor. AB - Changes in serum levels of calcium, magnesium, cAMP and monoamine oxidase (MAO) activity throughout pregnancy and during prolonged treatment for preterm labor with ritodrine were studied. cAMP levels in the third trimester of pregnancy were found to be significantly high and postpartum MAO activity was significantly low. In cases of preterm labor, serum levels of calcium, magnesium and MAO activity were significantly low during the initial stage of ritodrine administration, then recovered after more than 1 week of treatment despite the fact that given doses and serum levels of ritodrine remained constant and that the serum level of cAMP was still high. These results suggested that a control mechanism for MAO activity during pregnancy might exist due to an increase in catecholamine and that the beta 2-effect of ritodrine induced a lowering in serum calcium and magnesium levels. However, some homeostatic mechanism to regulate the ions might come into play later on the mechanism of down-regulation of ritodrine. PMID- 2553555 TI - Catecholamines do not induce fibrinolytic activity increase in cultured bovine endothelial cells. AB - This study has investigated the catecholamine involvement in the fibrinolytic modulation of cultured bovine aortic endothelial cells (BAEC). Adrenaline and isoproterenol, at concentrations ranging from 10 to 100 microM, were unable to modulate the fibrinolytic response of these cells. beta-Adrenergic binding studies using 3H-CGP 12177 as radioligand evidenced the presence of about 23,113 +/- 2,065 sites/cell, with a KD of 1.23 +/- 0.29 nM. Isoproterenol stimulated cAMP accumulation at concentrations ranging from 1 to 100 microM, with a maximal accumulation of 30 pmol/10(6) cell. Hence, in our experimental conditions BAEC, although possessing functional beta-adrenergic receptors, were unable to increase any fibrinolytic activity in response to catecholamines. PMID- 2553556 TI - [Effect of ethanol on synaptosomal membrane potential in the rat brain]. AB - The membrane potential of rat brain synaptosomes was depolarized by ethanol at concentrations of over 0.8-1.6%. This ethanol-induced depolarization was augmented by 4-aminopyridine (4-AP) or the gluconate- -medium, but not affected by tetrodotoxin, choline-Cl, NH4Cl, picrotoxin, ethacrynic acid, furosemide or SCN- -medium. The ethanol-induced depolarization was augmented by 4-AP in Cl- - or gluconate- -medium, but not in SCN- -medium. These findings suggest that ethanol depolarizes the synaptosomal membrane potential through potassium and anion-related mechanisms, probably through potassium channels. PMID- 2553557 TI - [5-Hydroxytryptamine receptors]. AB - The existence of two different functional receptors for 5-hydroxytryptamine (5 HT) was first proposed by Gaddum and Picarelli. Aided by the development of radioligand binding techniques, the heterogeneity of 5-HT receptors has become more apparent in the past ten years. There are three main types of 5-HT receptors: 5-HT1, 5-HT2 and 5-HT3. Moreover, 5-HT1 is heterogenous and can be divided into 5-HT1A, 5-HT1B, 5-HT1C and 5-HT1D subtypes. 5-HT1B is probably related to the 5-HT autoreceptor controlling 5-HT release. Multiple 5-HT receptors are differentially distributed throughout the brain, and the agonist receptor interaction is altered by physical parameters and chemicals, suggesting that the receptors may be physiologically relevant. Three 5-HT receptor subtypes, 5-HT1A, 5-HT1C and 5-HT2, have been cloned. All three receptors contain approximately 450 amino acids arrayed as seven transmembrane domains. 5-HT1 and 5 HT1A are coupled to adenylate cyclase positively and negatively, respectively, while 5-HT1C and 5-HT2 are coupled positively to phospholipase C. 5-HT1A is also coupled to the opening of K+ channels in hippocampal pyramidal cells. A number of 5-HT-induced physiological responses have been shown to correlate with the 5-HT receptor subtypes. Based on a number of pharmacological studies, it seems likely that the mode of action of certain psychotropic drugs is closely related to the activity of central 5-HT receptors. PMID- 2553558 TI - Effect of N,N'-bis(alkyldimethyl)-alpha, omega-alkanediammonium dibromides on bacteria of the genus Clostridium. AB - Antibacterial effect of 17 ammonium compounds of the type of N,N' bis(alkyldimethyl)-alpha, omega-alkanediammonium dibromides was tested on anaerobically sporulating bacteria of the genus Clostridium. A sizable antibacterial activity was displayed by five N,N'-bis(alkyldimethyl)-1,6 hexanediammonium dibromides and by four N,N'-bis(decyldimethyl)-alpha, omega alkanediammonium dibromides. These compounds exhibited activity higher than, or comparable with, that of the reference standards Ajatin and Septonex. The maximum antibacterial activity was found in compounds whose alkyl chain contained 9-12 carbon atoms. Compounds with a lower number of carbon atoms in the chain (less than 8) exhibited a low activity. PMID- 2553559 TI - [Viruses of the herpes group]. PMID- 2553560 TI - [Neurologic diseases in herpesvirus infections]. AB - Herpes viruses can give rise to numerous virological infections. Life-threatening situations arise with encephalitis caused by HSV or VZV. Zoster vasculitis is probably more frequent than its diagnosis would suggest, and usually results in paralysis. A particularly troublesome condition is post-herpetic neuralgia that requires a wearisome and often complex treatment. Cerebral nerve involvement and polyradiculitis may be associated with almost all the human pathogenic herpes viruses. In patients with herpetic CNS involvement, early treatment is necessary. PMID- 2553561 TI - [Virus latency in infections with herpesviridae. Herpes simplex and varicella zoster virus: pathogenesis, clinical consequences]. AB - Once the organism has been invaded, all herpesviruses persist. This is accomplished by a change in the acute productive phase of the infection into the latent form. Reactivation of the latent virus ensures renewed transmissibility of the infection. This pathogenetic principle is ensured by an extremely finely tuned virus-host interrelationship which has developed in a unique specific manner for each herpesvirus. Pathogenetic fundamentals are discussed taking herpes simplex and the varicella-zoster virus as examples, and the resulting clinical consequences are shown. PMID- 2553562 TI - [Herpes virus infections in immunosuppressed patients. Herpes simplex and varicella zoster virus: prevention and therapy]. AB - Herpes simplex virus (HSV) and varicella-zoster virus (VZV) giving rise to primary or recurrent infections still threaten immunocompromised patients. However, such prophylactic measures as passive and active immunisation against VZV infection, and antiviral chemoprophylaxis and chemotherapy of HSV- and VZV infections have improved the prognosis of these patients considerably. PMID- 2553563 TI - [The significance of herpesviridae in ophthalmology]. AB - Herpesviruses are the most common causes of infection-related loss of vision or blindness in our part of the world. A better understanding of the underlying pathophysiology, together with major pharmaceutical advances have led to medical and surgical therapy which enables us to preserve or restore useful vision in most patients. PMID- 2553564 TI - [AIDS: small, life-prolonging steps. Antiviral therapy is more efficient--a decrease in side effects]. PMID- 2553565 TI - [How can a female protect herself from HIV infection?]. AB - In view of the changing patterns within the population groups affected with AIDS, the need for preventing transmission of the disease during heterosexual genital intercourse is becoming increasingly important. The problem as to how the woman can protect herself without the help of her partner has so far received little attention. Safer sex is not always practicable. The fact that the woman can be infected by HIV during genital intercourse is not doubted. The use of vaginal foam tablets containing 75 mg nonoxinol 9, which have been used for contraception for many years, seems to be a valuable measure of protection. The virucidal threshold concentration for nonoxinol 9, established in numerous in vitro studies, is far exceeded in practical intravaginal use of the foam tablets. An additional protection of the woman may be afforded by the use of a vaginal diaphragm. PMID- 2553566 TI - The effects of captopril and enalapril on tissue levels of various elements in spontaneously hypertensive rats. AB - The effects of two inhibitors of angiotensin I converting enzyme, captopril and enalapril, on the concentrations of Na, K, Ca, Mg, Fe, S, P, Sr, Mn, Cu and Zn ions in blood, plasma, heart, skeletal muscle, liver and kidney of spontaneously hypertensive rats (SHR) were studied. Captopril and enalapril were given by the intraperitoneal route for 15 days, at 160 mumols/kg/d and 40 mumols/kg/d, respectively. Elements in tissues were determined by inductively coupled plasma emission spectrometry with a JY 48 instrument. The common changes produced with the two drugs were: a decrease of Na in muscle (-10%), a decrease of Ca in plasma and kidney (less than -10%) and a decrease of Mn in liver (-15%). The main effects observed with only one of the two drugs were: an increase of Cu in plasma (+26%) with captopril, and increases of Sr in heart (+56%), muscle (+79%) and liver (+74%) with enalapril. Zinc concentration in tissues was not modified, except for an increase in liver with captopril (+13%) and a decrease in heart with enalapril (-11%). PMID- 2553567 TI - Characteristics of the dopamine receptors involved in the anorectic effects of apomorphine in mice. AB - In food-deprived mice apomorphine injected SC induced a brief (15-30 min) dose dependent (30-150 micrograms/kg) reduction in food intake. This effect occurred in naive mice as well as in mice habituated to a food deprivation procedure. The anorectic effect of apomorphine (150 micrograms/kg SC) was antagonized by sulpiride (ID50 = 8.6 mg/kg) and by haloperidol (ID50 = 66 micrograms/kg) but domperidone was ineffective (250 micrograms/kg). Mice submitted to a semi-chronic (6 d) blockade of dopamine receptors by haloperidol or injected intracerebroventricularly with 125 micrograms 6-hydroxydopamine 21 d before testing failed to develop a hypersensitivity to the anorectic effect of apomorphine (60 micrograms/kg). Although a single apomorphine injection (5 mg/kg) induced tolerance to the hypothermic effect of a second apomorphine injection of 150 micrograms/kg, it did not modify the anorectic effect. Repeated apomorphine injection (5 x 5 mg/kg) resulted in a slight but significant reduction in apomorphine-induced anorexia. A similarly significant reduction was not observed in mice submitted to repeated injections of dexamphetamine (5 x 5 mg/kg). PMID- 2553568 TI - Pharmacokinetics and effects on exercise heart rate of PK 11195 (52028 RP), an antagonist of peripheral benzodiazepine receptors, in healthy volunteers. AB - PK 11195 (or 52028 RP; 1-(2-chlorophenyl)-N-methyl-N-(1-methylpropyl)-3 isoquinoline carboxamide), an antagonist of the peripheral-type benzodiazepine receptors which are coupled to calcium channels, was administered to 10 healthy volunteers in order to study the pharmacokinetics and cardiovascular effects of the drug. PK 11195 was randomly administered intravenously (10 mg) and orally in three single dosages (100, 200 and 400 mg). Placebo was only given orally. Heart rate and blood pressure were recorded at rest and during exercise tests which were performed at 0, 1, 3, 6 and 24 h after dosing on each study day. The results showed that after IV administration, PK 11195 was rapidly distributed in two or three open compartments. Its elimination T 1/2 was short (3.7 +/- 3.0 h) with high interindividual variability. After oral ingestion the pharmacokinetics of PK 11195 were linear over the range of 100-400 mg single oral doses with a stable absolute bioavailability (33%). T 1/2 elimination was prolonged (7-12 h) and the presence of secondary increases in plasma concentration at 8-10 h and 22-24 h after drug absorption may have been related to enterohepatic cycling. No unchanged PK 11195 could be detected in urine. PK 11195 did not significantly modify heart rate and blood pressure at rest or during exercise and was well tolerated by the subjects. These data suggest a high inter-individual variability in PK 11195 disposition with extensive metabolism in normal exercising volunteers. PMID- 2553569 TI - New hope in irresectable hepatoma? AB - Primary hepatocellular carcinoma (HCC) is mostly irresectable and the survival time is short. Treatment can be palliative but hardly improves survival. Some promising future aspects that should be considered include continuous intra hepatic chemotherapy (fraught with difficulties but nevertheless worth pursuing), direct percutaneous injection of alcohol into the tumor tissue under ultrasound guidance, as well as cryotherapy of "occult" tumors. Monoclonal technique may help to target toxic therapy to the tumors whereas monoclonal antibodies raised in humans may possibly obviate the imminent dangers of antibody formation that would be an obstacle. Although all this may still smack a little of "Zukunftsmusik" ("music of the future") that would require considerable perseverance and effort before it could be translated into reality, every avenue must of course be explored. Chemical techniques that give rise to new hopes feature radio-labelled lipids selectively retained in foci of hepatocellular carcinomas; research has already gone into the question of retention within the tumor microvasculature, within the extracellular space, and the uptake by tumor cells. The oily anticancer agent SMANCS has already been combined with ethiodol (Lipiodol), an iodine addition product of the ethyl esters of saturated and unsaturated fatty acids, and selectively infused, resulting in a reduction in tumor size, and the survival time also seems to be improved following the use of Lipiodol combined with SMANCS or with other cytotoxic agents. After an encouraging pilot study, we treated 16 patients suffering from HCC, with a combination of Lipiodol and epirubicin, a cytotoxic agent having an enhanced activity against HCC as compared with its parent drug doxorubicin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553570 TI - Validity of MR imaging for monitoring effects of percutaneous ethanol injection for HCC. AB - Magnetic resonance (MR) images of 44 hepato-cellular carcinoma (HCC) patients who had undergone percutaneous ethanol injection therapy (PEIT) were analyzed during and after the treatment. Using a long TE/TR spin-echo technique, the initial high intensity of the tumor decreased during treatment in all patients. Decrease in tumor intensity corresponded well with decrease of serum alpha-fetoprotein level and histopathological changes of the necrosis. During the long-term follow-up of three patients, the reappearance of a highly intense area was readily detected by MR imaging, and hence the recurrence of the tumor, as determined clinically or histopathologically, was confirmed. The long TE/TR spin-echo technique proves most helpful in monitoring the effectiveness of PEIT for HCC, as well as for its recurrence. PMID- 2553571 TI - Inhibition by a nonmetabolized analogue of L-leucine of protein biosynthesis in tumoral pancreatic islet cells. AB - The effect of L-leucine, its deaminated metabolite 2-ketoisocaproate and its nonmetabolized analogue b(+/-)2-aminobicyclo[2,2,1]-heptane-2-carboxylic acid (BCH) upon protein labelling was examined in tumoral islet cells (RINm5F line) exposed to L-[4-3H]phenylalanine or L-[3-3H]serine. The interpretation of the results, in terms of changes in biosynthetic activity, was obscured by a possible interference of the tested nutrients with the uptake and further metabolism of the tracer tritiated amino acids. Nevertheless, when the cells were preincubated with the nutrient secretagogues and then incubated in the sole presence of L-[3 3H]serine, BCH, but not L-leucine or 2-ketoisocaproate, still inhibited protein labelling, this coinciding with a decrease in the ratio between TCA-precipitable and total radioactivity in the RINm5F cells. The inhibitory action of BCH was antagonized, to a limited extent, by D-glucose. It is proposed that BCH could be used as a tool to interfere with the function and growth of insulinoma cells. PMID- 2553572 TI - Thyroid hormone action on ACTH secretion. AB - Thyroid hormone effects on pituitary ACTH have not been well established. Adult male Sprague-Dawley rats were rendered hypo- and hyperthyroid while undergoing treatment with 6-Propylthiouracil (PTU) and L-Thyroxine (T4). At the time of decapitation, plasma values for T4 (micrograms/100 ml) were 3.9 +/- 0.4 in the control, 17.3 +/- 2.2 in the T4 and less than 2 in the PTU treated group; plasma T3 and TSH confirmed hyper- and hypothyroidism in the T4 and PTU treated groups respectively. Plasma immunoassayable ACTH and corticosterone were significantly increased in hyperthyroid and decreased in the PTU treated animals. Pituitaries were removed and incubated in DMEM. After 3 h incubation, ACTH content and secretion to the medium were significantly lower in the PTU group. As expected, pituitary TSH content and secretion were decreased in the T4 treated animals. These data indicate that thyroid hormones influence pituitary-adrenal function by increasing ACTH secretion and consequently corticosterone production. PMID- 2553573 TI - Immunoreactive corticotropin-releasing hormone, adrenocorticotropin and cortisol in human plasma during pregnancy and delivery and postpartum. AB - The activity of the hypothalamo-pituitary adrenal axis was examined, by measuring the levels of immunoreactive (IR) corticotropin-releasing hormone (CRH), adrenocorticotropin (ACTH) and cortisol (F) in human plasma during normal pregnancy and after delivery with or without complications and during normal postpartum using a specific RIA. The level of IR-CRH in maternal plasma increased progressively during pregnancy, increased further at delivery and declined rapidly to the non-pregnant level on the 1st day postpartum. The level of IR-F in maternal plasma also increased progressively during pregnancy, increased further at delivery, but decreased slowly postpartum, not returning to the non-pregnant level within 5 days. Significant correlations were found between the level of IR CRH and IR-ACTH, IR-CRH and IR-F, and IR-ACTH and IRF in maternal plasma both during pregnancy and after delivery. It is noteworthy that the concentration of IR-CRH in the maternal plasma at delivery was higher in multiple pregnancy than in normal pregnancy, and that the level of IR-CRH in the umbilical cord in uncomplicated cases was much lower than that in the maternal plasma, and was significantly lower than those in the umbilical cord plasma in cases of asphyxia, IUGR or premature delivery. The level of IR-F, not IR-CRH and IR-ACTH, at normal vaginal delivery was significantly higher than that at elective cesarean section. On these results, we investigated the feto-maternal-hypothalamo-pituitary adrenal axis during pregnancy and delivery, in which CRH plays an important role. PMID- 2553574 TI - Tc-99m-(V)-DMSA: the new sensitive and specific radiopharmaceutical for imaging metastases of medullary thyroid carcinomas? AB - In the follow-up of five patients with histologic proven medullary thyroid carcinoma (MTC) and raised serum calcitonin and CEA levels the pentavalent Tc-99m (V)-DMSA and the Tc-99m-MDP bone scan had the highest sensitivity in the localisation of metastases. Both methods are not tumor specific. A false positive Tc-99m-(V)-DMSA uptake in an old osteomyelitis of one vertebra could be demonstrated. The J-123-MIBG and In-111-F(ab2)' antibody scan did not allow to localise one of the above described metastases. In conclusion in the follow-up of patients with MTC and elevated tumor marker concentrations the Tc-99m-(V)-DMSA and the Tc-99m-MDP bone scan should be the second diagnostic procedures after sonography has been performed. PMID- 2553575 TI - Tumor necrosis factor alpha production by peripheral blood mononuclear cells of patients with chronic liver disease. AB - We investigated the production of tumor necrosis factor alpha by peripheral blood mononuclear cells of patients with chronic liver disease and its association with hepatitis activity. Tumor necrosis factor alpha production was measured with an enzyme-linked immunosorbent assay. Tumor necrosis factor alpha production by peripheral blood mononuclear cells stimulated with recombinant gamma-interferon of patients with chronic active hepatitis (5.8 +/- 4.0 units per ml, p less than 0.05) and patients with cirrhosis (4.1 +/- 2.1 units per ml, p less than 0.05) was significantly increased when compared with controls (2.5 +/- 1.6 units per ml). Tumor necrosis factor alpha production by peripheral blood mononuclear cells stimulated with a combination of recombinant gamma-interferon and recombinant interleukin 2 of patients with chronic persistent hepatitis (5.8 +/- 3.8 units per ml, p less than 0.05), patients with chronic active hepatitis (8.9 +/- 3.0 units per ml, p less than 0.001) and patients with cirrhosis (6.7 +/- 3.2 units per ml, p less than 0.05) was significantly increased in comparison with controls (3.3 +/- 1.8 units per ml). Tumor necrosis factor alpha production of patients with chronic active hepatitis was significantly higher than that of patients with chronic persistent hepatitis (p less than 0.05). There was a significant correlation (r = 0.5699, p less than 0.005) between tumor necrosis factor alpha production and histologic activity index in patients with chronic persistent hepatitis or chronic active hepatitis. These findings show that tumor necrosis factor alpha production is increased in chronic liver disease and that the increased tumor necrosis factor alpha production is related to hepatitis activity. PMID- 2553576 TI - Primary intestinal lymphoma of Ki-1 large cell anaplastic type with mesenteric lymph node and spleen involvement in a renal transplant recipient. AB - A laparotomy performed for an occlusive syndrome, disclosed the presence of an ileal lymphoma in a renal graft recipient, 17 years after a first renal transplant. Mesenteric lymph node and spleen involvement were associated. The morphology of the large cells, the type of infiltration of the lymph node simulating metastatic carcinoma or malignant histiocytosis were highly suggestive of a new type of high grade malignancy lymphoma recently described which is called 'large cell anaplastic ML'. The immunolabelling study confirmed this diagnosis, demonstrating the following phenotype CD4, CD25 (TAC), CD30 (Ki-1), OKT 9 (transferrin receptor), EMA and HLA DR positivity. The tumour cells were negative for the other T, B and histiomonocytic markers. This case, of a large cell anaplastic Ki-1 positive lymphoma, expressing CD4, in a renal graft recipient shows that high grade ML in such patients are not all of the B cell type. PMID- 2553577 TI - Paneth cell-rich carcinoma of the stomach. PMID- 2553578 TI - Use of enzyme histochemistry in the diagnosis of pancreatoblastoma. AB - The macroscopic, light microscopic and ultrastructural findings in a case of pancreatoblastoma in a neonate are presented. No evidence for exocrine or endocrine secretion could be found by histochemistry, immunocytochemistry or electron microscopy. However, enzyme histochemical techniques to localize activity of 'glucose-6-phosphatase', acid phosphatase, esterase and esteroprotease supported both a pancreatic origin for the tumour and a biphasic pattern of differentiation. PMID- 2553579 TI - Nephroblastomatosis of combined superficial and intralobar type with blastema predominant Wilms' tumour: report of a case. PMID- 2553580 TI - Human papillomaviruses and anal cancer. PMID- 2553582 TI - Research opportunities at the National Institute of Mental Health. PMID- 2553581 TI - Anal condylomas in men. 1. Histopathological and virological assessment. AB - A series of 128 biopsy specimens from anal condylomas in 73 homosexual or bisexual and 38 heterosexual men (mean (SD) age 31.8 (9.6) years) were subjected to histological assessment and human papillomavirus (HPV) typing by in situ DNA hybridisation with 35S-labelled HPV 6, 11, 16, 18, 31, and 33 probes. Most patients were also tested serologically for antibodies to human immunodeficiency virus (HIV). As evaluated on light microscopy, most (74%, 95/128) of the lesions were exophytic (papillary) acuminate warts, 15% (19) were flat, and 11% (14) were pigmented papulous lesions. No signs of anal intraepithelial neoplasia (AIN) were seen in 70% (90) of the 128 biopsy specimens (NAIN), 27% (35) were classified as showing AIN I, and another 2% (three) as AIN II. AIN was significantly (p less than 0.05) more often associated with papulous lesions, only 43% (6/14) of which showed NAIN compared with 72% (68/98) of acuminate condylomas. The duration of disease was directly related to the presence and severity of AIN in the lesions; thus in 47 lesions that had been present for more than 12 months, NAIN was found in 31 (66%), AIN I in 14 (30%), and AIN II in two (4%). HPV DNA of at least one of the six types tested for was detected in 109/125 (87%) lesions. HPV 6 and HPV 11 were the two most common types, comprising 57% (62) and 37% (40), respectively, of the 109 HPV DNA positive cases. Only seven (6%) biopsy specimens were associated with any of HPV types 16, 18, 31, or 33, which carry a high risk of potential malignant transformation. No association was found between sexual preferences of patients and the incidence of any of the various HPV types. Neither did the distribution of the various HPV types differ between men with antibody to HIV and those without antibody. All the men with antibody to HIV were homosexual or bisexual. On microscopy, 93% (38) of 41 lesions containing HPV 11 and 75% (48/64) of HPV 6 lesions were of the acuminate wart type; in comparison, the remaining 16 HPV 6 lesions were equally either flat or papulous (eight, 13% each). Of the 64 HPV 6 and 41 HPV 11 associated lesions, 73% (47) and 63% (26), respectively, were classified as NAIN. Only two lesions were associated with HPV 16, and both showed mild dysplasia. On the other hand, two HPV 6 induced lesions were associated with AIN II. No differences were found between HPV 6 and HPV 11 in duration of disease; (39%, and 27% respectively, had been present for more than 12 months). The results showed that overt anal wart disease was associated with HPV types 6 and 11 in most cases. Although HPV types considered as being of higher oncogenic potential were detected relatively rarely, the associated AIN in a relatively high proportion (31% 32/105) of HPV 6 or 11 induced lesions indicated that a malignant potential, even for HPV 11 associated anal warts, cannot be excluded. PMID- 2553583 TI - Unequal expression of complement C4A and C4B genes in rheumatoid synovial cells, human monocytoid and hepatoma-derived cell lines. AB - C4A and C4B are closely related homologous complement proteins encoded in the class III region of major histocompatibility complex (MHC). The regulation of their expression is under genetic and hormonal control. In this study we investigated the synovial fluid plasma ratio of C4A and C4B of rheumatoid (RA) and osteoarthritis (OA) patients, and a predominance of the C4B gene expression by the synovial macrophages of RA patients was demonstrated. To clarify the tissue specificity of the expression of C4A and C4B genes, human monocytoid cell line U937 and hepatoma-derived HepG2 cells were studied. The gene expression of C4A and C4B were markedly different in these cells since a relative predominance of C4B mRNA in U937 cells and excess of that of C4A in HepG2 cells were detected. Recombinant interferon-gamma (IFN-gamma) up-regulated the expression of C4A gene in both cells, but had apparently no effect on the C4B gene. Our results demonstrate dissimilar expression patterns for the two human C4 genes, suggesting different tissue specific regulation of human C4A and C4B. PMID- 2553584 TI - Proliferative T-cell response to glycoprotein B of the human herpes viruses: the influence of MHC and sequence of infection on the pattern of cross-reactivity. AB - Oligopeptides of the highly conserved herpes virus glycoprotein B (gB) were expressed from DNA fragments of the EBV gB (BALF4) and HSV-2 gB open reading frames as fusion proteins with the lambda CII protein and beta-galactosidase (GZ), respectively, in Escherichia coli. After immunopurification using anti-gB or anti-GZ affinity columns, the fusion proteins were used in vitro to stimulate human peripheral blood lymphocytes (PBL) or murine lymph node cells that have been primed with EBV, HSV-1, HSV-2, VZV or HCMV (all human herpes viruses) to proliferate. Results obtained in BALB/c mice indicate that different herpes viruses induce different levels of T-cell response to each other and to gB, over a range of type-specific and cross-reactive T-cell epitopes. There is a lack of correlation of immunogenicity and antigenicity in the generation of T-cell responses between some of the viruses. Major T-cell epitopes are located at the C terminal half of the gB molecule. The T-cell response to gB in healthy individuals seropositive for various combinations of the five herpes viruses differed markedly from individual to individual, even when they are seropositive to the same set of herpes viruses. However, two individuals with high proliferative T-cell response to VZV and sharing HLA A2, B7, DR2 and DQw1 are also good responders for cross-reactive gB/fragments and for virus antigen of all the five herpes viruses. Therefore the data obtained demonstrated that the MHC and the immune interaction arising from cross-reactive T-cell response evoked by other herpes viruses may determine the pathogenesis of a herpes virus infection. PMID- 2553585 TI - Different effect of prostaglandin E2 on B-cell activation by two distinct B-cell differentiation factors, B151-TRF1/IL-5 and B151-TRF2: selective inhibition of B151-TRF2-induced antibody response through increases in intracellular cyclic AMP levels. AB - Effects of prostaglandin E2 (PGE2) on murine B-cell activation induced by two distinct B-cell differentiation factors, B151-TRF1/IL-5 and B151-TRF2, were examined. A final differentiation of unprimed B cells into IgM-producing cells induced by B151-TRF2 was markedly inhibited by PGE2 at physiological concentrations (around 10(-8) M), whereas B151-TRF1/IL-5- induced antibody responses of unprimed as well as activated B cells were not affected by PGE2, even at 10(-6) M. B-cell responses induced by B151-TRF2-like factors from autoimmune-prone MRL/lpr mice were also inhibited by PGE2. Biphasic increases in intracellular cyclic AMP (cAMP) levels were induced by culturing B cells with 10( 6) or 10(-8) M PGE2: rapid increases within 8 min and delayed increases around 16 hr. The direct addition of dibutyryl cAMP to cultures of B cells resulted in marked inhibition of antibody responses when stimulated with B151-TRF2 but not with B151-TRF1/IL-5. The B151-TRF2-induced antibody responses were also inhibited by cAMP-elevating reagents such as forskolin, cholera toxin and theophylline. Furthermore, 2'.5'-dideoxyadenosine, which is an inhibitor of adenylate cyclase, prevented the PGE2-mediated cAMP accumulation in unprimed B cells as well as the PGE2-mediated inhibition of B151-TRF2-induced B-cell responses when added at the initiation of culture. These results suggest that PGE2 inhibits B151-TRF2-induced antibody responses through the activation of adenylate cyclase and subsequent accumulation of intracellular cAMP, whereas B151-TRF1/IL-5-responsive B cells are resistant to the inhibitory effect of PGE2 and cAMP. PMID- 2553586 TI - Effect of sodium fluoride on the generation of lipoxygenase products from human polymorphonuclear granulocytes, mononuclear cells and platelets--indication for the involvement of G proteins. AB - Incubation of human polymorphonuclear granulocytes, monocytes and platelets with sodium fluoride (NaF) results in a time- and dose-dependent generation of leukotrienes and 12-HETE, respectively. This release was not influenced by pretreatment with pertussis toxin or cholera toxin. The mediators are detectable after a lag phase of about 5-10 min. Inactivation of LTB4 by the neutrophils via omega-oxidation into 20-hydroxy-LTB4 and 20-carboxy-LTB4 is inhibited by NaF. In combination with other cell stimuli, NaF showed modulatory effects, such as an enhanced formation of the leukotrienes when FMLP, opsonized zymosan, PMA, and arachidonic acid were applied as stimuli. Prestimulation of cells with NaF causes an increased [3H]guanylylimidodiphosphate binding to isolated membrane preparations, indicating an enhanced exchange rate for GDP to GTP. Our data demonstrate that a direct activation of GTP-binding proteins results in the generation of the inflammatory mediators and provides evidence for the involvement of the signal-transduction pathway. PMID- 2553587 TI - Autocrine regulation of 1,25-dihydroxycholecalciferol metabolism in myelomonocytic cells. AB - In this study the effects of vitamin D metabolites on the myelomonocytic precursor cell line U937 have been compared with those of phorbol myristate acetate (PMA). PMA was used as a cell modulating agent in order to avoid effects of binding of the exogenous vitamin D metabolites receptors within the cell, which would interfere with subsequent measurement of these receptors in studies of the vitamin D3 metabolic pathway. Both the active 1,25 DHCC form of vitamin D3 and the inactive 24,25 DHCC metabolite inhibit cell proliferation and induce 24 hydroxylase activity, but not 1 alpha-hydroxylase activity. These effects are dose-dependent and maximum enzyme activity is seen in the adherent cell population, which is induced by these compounds. PMA inhibits proliferation of U937 and increases receptors for 1,25 DHCC in these cells (like the vitamin D metabolites). However, unlike the vitamin D metabolites, PMA induces 1 alpha hydroxylase activity rather than 24-hydroxylase activity. Thus, while PMA and 1,25 DHCC have some similar effects on monocyte precursor cell line differentiation, there is a difference between the effects of the two agents on the vitamin D metabolic pathway. The former promotes synthesis of the active metabolite, and the latter induces an enzyme which renders the metabolite inactive. If these results are considered together, they are consistent with the hypothesis that 1,25 DHCC has an autocrine role within the mononuclear phagocyte system. PMID- 2553588 TI - Foot-and-mouth disease virus-neutralizing antibodies induced in mice by anti idiotypic antibodies. AB - A neutralizing monoclonal antibody (nmAb) to foot-and-mouth disease virus (FMDV) was used as antibody-1 (AB1) to induce anti-idiotypic antibodies (a-IdAb) in rabbits. The rabbit a-IdAb (AB2) were isolated on protein A-Sepharose, followed by cycles of separation on idiotype and isotype affinity columns. The specificity of the AB2 for the paratope of AB1 was determined by direct binding to AB1 in solid-phase radioimmunoassay (SP-RIA), and by competition RIA (C-RIA) with virus for binding to the AB1. The AB2, termed a-2PD11, was utilized to immunize six groups of female Swiss mice at weekly intervals with either one of three formulations, in doses of 50 micrograms or 5 micrograms, given in single subcutaneous (s.c.) spots. Anti-viral antibody (AB3) was first detected by RIA at the fifth week in the 50 micrograms/dose groups, and maximum levels were reached at the sixth week in the 50 and 5 micrograms/dose groups. The AB3 levels were at least three times higher for mice given 50 micrograms doses. In addition, the AB3 were also shown to neutralize FMDV infectivity in tissue culture and in a suckling mouse protection assay. Overall, mice exhibited variable responses to immunization with AB2. In a subsequent trial, mice received multispot s.c. and footpad injections of 50 micrograms of a-2PD11 coupled to keyhole limpet haemocyanin (KLH) on a weekly basis. In these mice, AB3 was detected earlier than in mice immunized with single s.c. injections. These results support the use of a IdAb as potential surrogates of critical determinants for FMD vaccines. PMID- 2553589 TI - Effect of muramyl peptides and tumor necrosis factor on oxidative responses of human blood phagocytes. AB - Adjuvant muramyl dipeptides enhanced the intracellular oxidative burst induced by phorbol myristate acetate in purified human polymorphonuclear (PMN) cells and in monocytes. A stronger priming effect was obtained when muramyl dipeptide was conjugated to a protein carrier. Recombinant human tumor necrosis factor (TNF) did not modify the level of intracellular NBT reduction in PMA-stimulated PMN, although it slightly increased the secretion of superoxide anion. In contrast, TNF enhanced the intracellular oxidative burst of monocytes even at the concentration of 10 pg/ml. In human PMN only, the combination of TNF and muramyl dipeptide induced a higher oxidative response than each stimulant alone. PMID- 2553590 TI - Current status of pharmacological receptors in mammalian central nervous system. PMID- 2553592 TI - Effect of dexamethasone and reserpine on brain synaptosomal phosphorylation in lithium treated rats. AB - Increase in brain synaptosomal protein phosphorylation after lithium treatment was unaffected by prior treatment with dexamethasone, but decreased after reserpine administration indicating the involvement of neurotransmitters in lithium induced increase in synaptosomal phosphorylation. PMID- 2553591 TI - Gonadal toxicity of short term chronic endosulfan exposure to male rats. AB - Endosulfan was studied for its effect on rat testicular toxicity in relation to the enzymes of androgen biosynthesis, viz. 3 beta-hydroxysteroid dehydrogenase (EC 1.1.1.145, 3 beta-HSD) and 17 beta-hydroxysteroid dehydrogenase (EC 1.1.1.64, 17 beta-HSD); cytosolic conjugation enzyme, glutathione-S-transferase (EC 2.5.1.18); and testicular as well as serum testosterone levels at the dose levels of 2.5, 5.0, 7.5 and 10 mg/kg body weight fed orally for 7 and 15 days. Organ and body weights of the treated animals did not change significantly, however, the testicular protein contents were found to be increased appreciably after 7 days treatments. The activity profile of cytosolic conjugation enzyme showed much remained low during 7 days treatment, however, the two steroidogenic enzymes showed much individual variations in response to endosulfan treatments. An overall varied response with respect to testosterone biosynthesis and its secretion to serum was observed suggesting nevertheless, a profound hormonal imbalance caused by this insecticide to male gonads on short term chronic exposures. PMID- 2553593 TI - Histomorphology of Wilms' tumour. AB - Eleven cases of Wilms' tumour received at Banglore Medical College, Bangalore are being studied focussing on the histopathological appearance of these tumours. One case exhibiting neuronal and glial differentiation, which is a rare observation, was noticed and is reported. The clinicopathological features and a brief review of literature are being presented. PMID- 2553594 TI - Association of hepatitis B virus with chronic liver diseases and hepatocellular carcinoma. AB - The idea of hepatitis-cirrhosis-hepatocellular carcinoma sequence in liver was proposed by the workers in tropical Africa, the homeland of hepatocellular carcinoma. The discovery of Australia antigen by Blumberg et al provided the missing link and it was observed by several workers as well as the present group that Hepatitis B surface antigen (HBSAg) and that to Hepatitis B Virus (HBV) is someway related with the incidence of chronic liver disease and hepatocellular carcinoma (HCC). PMID- 2553595 TI - Adenoid cystic carcinoma (ACC) of left sphenoidal sinus (case report). PMID- 2553596 TI - Pre-cancerous lesions of stomach. AB - Total 39 cases of carcinoma stomach were noticed out of 142 malignant tumours of GIT (27.46 percent). Histologically maximum cases were of diffuse type (56.41 percent) followed by intestinal type (35.89 percent) and indolent mucoid carcinoma (7.69 percent) of the stomach. The surrounding epithelium showed lot of changes in the intestinal type of carcinoma stomach. About 78.57 percent showed intestinal metaplasia, 14.28 percent of these cases showed chronic gastric ulcer and severe dysplasia (carcinoma in situ) and another 14.28 percent revealed villous adenoma with carcinoma in situ. In contrast to this, in diffuse variety, only 13.63 percent cases revealed intestinal metaplasia, 27.27 percent showed basal cell hyperplasia, stratification of the epithelium of crypts and diffuse infiltration of mucosa by malignant cells and 4.54 percent showed atrophic gastritis also. In mucoid carcinoma all cases had basal cell hyperplasia and stratification of crypts. Hence these conditions should be taken as premalignant lesions of stomach and should be cured in proper time. PMID- 2553597 TI - Acutely transforming retroviruses as carcinogens in mouse embryos. AB - Spontaneous and induced transmission of oncogenic retroviruses from parents to offspring is well documented in mammals, as exemplified by murine leukaemia viruses. In addition, it is now common knowledge that transgenic mice, carrying viral and cellular transforming genes, can show unusual tumour incidences and can transmit this character to successive generations. The present report deals with the occurrence of tumours in rodents exposed directly in utero to murine sarcoma viruses (MSV). Attention was focused on whether MSV are involved in the etiopathogenetic processes leading to early-life and adult-type neoplasms in mice. According to our data, this is a likely possibility, since, for example, Kirsten MSV (Ki-MSV) induced stage-specific tumours in CD-1 mice, including lung tumours and skin papillomas, following intraembryonal injections on gestation days 8 and 10, respectively. When the injections were given in the second half of pregnancy, however, only capillary angiomas and other vascular malformations of the brain, as well as mesenchymal sarcomas, were induced in the newborn. Harvey MSV (Ha-MSV), a close relative of Ki-MSV, and Moloney MSV (Mo-MSV) were much less dependent on the stage of embryogenesis, and mesenchymal sarcomas were frequently detected in prenatally infected animals. Other MSV viruses, including 3611, J-2 and 4070A, were active carcinogens in a way totally independent of the embryonal stage exposed, and tumours in the progeny ranged from mesenchymal sarcomas to lymphoblastomas and rhabdomyosarcomas, respectively. We next looked at the possibility that spontaneous tumours of CD-1 mice might result from the interaction of certain chemical carcinogens with developmentally expressed components of the Ki-MSV virus. The experiments conducted thus far have provided dubious results. An example is given by experiments with N-ethyl-N-nitrosourea (ENU). This compound is a powerful transplacental carcinogen in CD-1 mice, where it induces lung tumours. It was expected that by exposing developing mice to Ki MSV on day 8 of embryogenesis and to ENU late in pregnancy or in newborns, a great increase in the incidence of lung tumours would have appeared in the offspring. However, this did not happen, and the agents behaved as if different cellular targets were involved. Understanding this difference could be very informative and useful in identifying neoplastic factors in mammalian embryos. PMID- 2553599 TI - Genetic and molecular aspects of human multigenerational carcinogenesis. AB - The notion that cancer is a genetic disease has gained increasing credence from the now numerous studies in which specific alterations of chromosome and of gene structure and activity in a variety of cancers have been identified, and, in particular, with the increasing awareness of the heritability of factors that predispose to the development of early, specific cancers in the offspring of parents carrying these predisposing genes. That the inheritance of a single allele is a major causal factor in certain childhood cancers, e.g., retinoblastoma and Wilms' tumour, has long been known, and the locations of the genes predisposing to these tumours have been mapped and at least one has been isolated. Over the past year, using genetic analysis, a further five different loci have been mapped that are involved in five other inherited cancer predispositions. Moreover, in most of these cases the evidence again suggests that one of the pair of alleles at the locus linked to the predisposition undergoes somatic mutation (loss) within the cells that give rise to each of these tumours. A major factor in the origin of these tumours therefore is a loss of a tumour suppressing role of the loci in question. At the present time, seven such suppressor loci on six different chromosomes have been identified, and in a number of cases evidence has been presented for the occurrence of other specific genetic changes that may be necessary for the emergence of a neoplasm. Specific gene or chromosome loss is, however, not confined to cancers associated with the inheritance of a single gene but is increasingly being observed in various more common sporadic cancers, and these data are reviewed and some new data presented. Studies on sporadic cancers have revealed a wide range of different oncogenes in which mutation or abnormal regulation is a cardinal factor leading to a neoplastic state. The products of many of these oncogenes have homologies to growth factors, growth factor receptors or are signal or DNA binding proteins, and specific mutations have been characterized in a number of e-oncogenes in neoplastic human cells. In contrast to genes which may suppress neoplasia, the c oncogenes act as positive factors, although the presence of a single mutated oncogene is not sufficient on its own to result in neoplastic transformation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2553598 TI - Comparative transplacental carcinogenesis by directly acting and metabolism dependent alkylating agents in rodents and nonhuman primates. AB - Transplacental carcinogenesis by N-ethyl-N-nitrosourea (ENU) was studied in patas (Erythrocebus patas) and rhesus (Macaca mulatta) monkeys. Repeated intravenous injections throughout pregnancy caused gestational choriocarcinoma in female patas monkeys and a variety of non-trophoblastic neoplasms in their offspring. Latent periods for transplacentally induced tumours varied from less than one month to more than ten years. One case of congenital neoplasm was observed. Certain kinds of neoplasms were observed only in transplacentally exposed offspring, and not in monkeys given the same carcinogen during juvenile or adult life. These included a variety of embryonal tumours, especially nephroblastoma, and tumours of the brain, mostly gliomas. Schwannomas of the peripheral nervous system were not observed in patas monkeys given ENU. One embryonal tumour yielded DNA that transformed NIH 3T3 cells in a transfection assay. Similar protocols performed in rhesus monkeys also yielded a variety of tumours in the offspring, including brain tumours; but in this species the most common embryonal tumour was pulmonary blastoma, and no choriocarcinoma was seen in adult females given ENU during pregnancy. Administration of N-nitrosodiethylamine to patas monkeys during pregnancy at first appeared to have had no effect on the offspring, but administration of phenobarbital beginning at four years of age resulted in the rapid appearance of multiple hepatocellular tumours. With regard to potential human risk from prenatal exposure to carcinogens, three conclusions deserve special emphasis: (1) the extreme susceptibility of the fetal primate central nervous system to certain chemical carcinogens, which confirms and reinforces what was previously known from studies in rodents; (2) the prolonged latency of transplacentally initiated epithelial tumours and the importance of subsequent postnatal exposure to promoting agents; and (3) the concurrent risk of transplacental chemical carcinogenesis in offspring and gestational choriocarcinoma in the mother, suggesting that gestational choriocarcinoma with its short latent period may serve as an epidemiologically exploitable marker for human populations in which transplacental carcinogenesis is likely. PMID- 2553600 TI - Expression of beta-adrenergic receptors by astrocytes isolated from adult rat cortex. AB - Astrocytes are the most numerous cell type in the central nervous system. When cultured, these cells express a wide variety of receptors for neurotransmitters. Nonetheless, it has yet to be firmly established that adult astrocytes in situ express receptors for neurotransmitters. In this report the technique of combined receptor autoradiography and immunocytochemistry has been applied to isolated astrocytes from adult animals to examine beta-adrenergic receptor (beta-AR) expression by these cells. It is shown that fibrous and protoplasmic astrocytes isolated from adult rat cerebral cortices specifically bind the beta-AR ligand iodocyanopindolol and that this binding is inhibited by the beta-AR ligands propranolol and isoproterenol. These results indicate that at least two of the major morphological subtypes of adult astrocytes express beta-ARs. PMID- 2553601 TI - Sodium channels in astrocytes of rat optic nerve in situ: immuno-electron microscopic studies. AB - Immuno-electron microscopic localization of sodium channels within astrocyte somata and processes of adult rat optic nerve was demonstrated with polyclonal antibody 7493. In immunoblots of crude glycoproteins from adult rat optic nerve, antisera 7493, which is directed against purified rat brain sodium channels, recognizes a 260 kDa protein. Antisera 7493 intensely immunostains axon membrane at nodes of Ranvier. Associated perinodal astrocyte processes are also stained with antisera 7493. In addition, astrocyte cell bodies and major processes exhibit immunoreactivity with antibody 7493. Immunostaining with antisera 7493 is heterogeneously distributed within astrocyte cytoplasm and also appears to be associated with some regions of astrocyte plasmalemma. Glial filaments are not immunostained with 7493 antisera. Astrocyte processes forming the glial limitans and surrounding blood vessels display reduced immunoreactivity to 7493 compared to longitudinally oriented or perinodal astrocyte processes. However, some focal regions of the glial limitans exhibit robust 7493 immunostaining. Oligodendrocytes do not display 7493 antisera immunoreactivity. Optic nerve sections incubated with preimmune sera or with 7493 antisera that had been previously adsorbed with purified sodium channel protein, exhibited no immunoreactivity. These results demonstrate localization of sodium channels within astrocytes in situ of rat optic nerve and extend previous electrophysiological and pharmacological findings of sodium channels in cultured astrocytes. Possible functional roles of sodium channels within astrocytes are discussed. PMID- 2553603 TI - C1q binding to human vascular smooth muscle cells mediates immune complex deposition and superoxide generation. AB - Evidence was obtained for the binding of C1q to the membrane of cultured vascular smooth muscle cells derived from human umbilical cord veins. C1q was fixed to the cell membrane at 4 degrees C, whereas it was ingested into the cytoplasm, as a cytoplasmic inclusion, when tested at 37 degrees C. The addition of C1q in advance inhibited the subsequent binding of C1q. Neither fibronectin nor laminin was detected on the cell membrane. Aggregated IgG bound to vascular smooth muscle cells in the case of preincubation with C1q at 4 degrees C, whereas aggregated IgG did not bind to the cells in the absence of C1q. The addition of C1q molecules to the cells in suspension enhanced superoxide generation by vascular smooth muscle cells. There was no effect of C1q on superoxide generation by the cells in monolayer. These results suggest that C1q binds on the membrane of vascular smooth muscle cells via its specific receptor that mediates immune complex binding to the cells and superoxide generation. These properties elucidate the mechanisms by which circulating immune complexes deposit in the vascular wall, and subsequent degradation of tissue components surrounding vascular smooth muscle cells occurs through oxidative burst of the cells. PMID- 2553602 TI - In vivo and in vitro HeNe laser effects on phagocyte functions. AB - The goal of this work was to evaluate the effect of helium-neon (HeNe) laser irradiation on immunocompetent cells. We used the in vivo skin window method and in vitro granulocyte function tests. The study of cellular migration showed a marked decrease in vitro and in vivo in a dose-independent manner. Superoxide release was not modified by laser irradiation. The granulocyte's aggregation, when using PHA and PMA, presented a reduction that was statistically very significant, not as a subordinate dose. An increase of the release of ATP was demonstrated only at 4 joules and precedes granulocyte aggregation. When using Ca2+ ionophore A23187 as stimulus, laser irradiation at 1, 2 or 4J did not show any modification of granulocyte aggregation. The monoclonal antibody 60.1, which identifies a membrane antigen fundamental for aggregation and chemotaxis, is expressed in normal amounts on granulocyte membranes both before and after irradiation with a HeNe laser. In fact, Laser irradiation preferentially attacks the area of the cellular centrosome that determines a modification of cellular morphology. The electron microscope and immunofluorescence study with a monoclonal antibody have pointed out a disorganization of the microtubules. The alteration of some of the granulocyte functions is correlated to the damage in the centrioles. The granulocyte mitochondrial system and surface membrane remain intact, and this explains the normal production and release of free radicals. Further experiments are necessary to evaluate the clinical application of lasers in various diseases with immunophagocytic pathogenesis. PMID- 2553604 TI - Decreased opsonic activity for Staphylococcus aureus in neonatal and late gestation maternal sera. AB - Heat-killed Staphylococcus aureus (ATCC strain 25923) and Escherichia coli K-12 were used as target microorganisms for opsonization by serum from neonates or mothers at various stages of pregnancy or postpartum. The level of opsonic activity was evaluated by titrating serum for the ability to coat bacteria for recognition by normal human PMN. Recognition of the organisms was quantitated by measuring the PMN superoxide anion generation response to opsonized organisms. Studies show that opsonic activity for S. aureus was markedly decreased in serum from mothers in their second and third trimesters of pregnancy, in cord blood, and from infants at two weeks of age. Decreased activity was not observed in maternal serum in the first trimester of pregnancy and returned to control levels shortly after delivery. No significant difference was seen in opsonic activity for E. coli in these sera. The reduction in serum opsonic activity in mothers during the second and third trimesters of pregnancy suggests a systemic suppression of maternal immune responses during fetal development. This decreased activity is also observed in neonatal sera. PMID- 2553605 TI - Effect of a leukotriene B4 receptor antagonist on leukotriene B4-induced neutrophil chemotaxis in cavine dermis. AB - Leukotriene B4 (LTB4) is a proinflammatory product of arachidonic acid metabolism that has been implicated as a mediator in a number of inflammatory diseases. When injected intradermally into the cavine. LTB4 elicits a dose-dependent immigration (chemotaxis) of neutrophils (PMNs) into the injection sites as assessed by the presence of a neutrophil marker enzyme myeloperoxidase. SC-41930 (7-[3-(4-acetyl 3-methoxy-2-propylphenoxy)propoxy]-3,4-dihydro-8-p ropyl-2H- 1-benzopyran-2 carboxylic acid), a potent LTB4 receptor antagonist inhibited the chemotactic actions of LTB4 when coadministered into the dermal site and when given intravenously or orally with ED50 values of 200 ng, 0.5 mg/kg, and 0.6 mg/kg respectively. This compound may well have application in disease states, such as inflammatory bowel disease and psoriasis, where LTB4 is implicated as a proinflammatory mediator. PMID- 2553606 TI - Nutritional therapy of irritable bowel syndrome. AB - Nutritional factors relative to IBS include diagnostic and therapeutic considerations. Etiologically, foods do not cause IBS. A small percentage of patients with childhood allergic diatheses, usually in association with atopic dermatitis and asthma, may be intolerant to one or more of wheat, corn, dairy products, coffee, tea, or citrus fruits. Diagnostically, many patients labeled as IBS subjects are in fact intolerant to the ingestion of lactose-containing foods, sorbitol, fructose, or combinations of fructose and sorbitol. A precise dietary history will characterize this group. Taken in its broadest context, IBS involves the entire hollow tract inclusive of esophagus, stomach, small bowel, and colon. The symptomatic presentation relative to the hollow organ involved allows the selection of dietary manipulations that may help to reduce symptoms. Gastroesophageal reflux, a consequence of low LES pressure in some IBS patients, may be treated with the elimination of fatty foods, alcohol, chocolate, and peppermint. Delayed gastric emptying may be helped by the elimination of fatty foods and reduction of soluble fiber. Aberrant small bowel motor function may be ameliorated by reduction of lactose, sorbitol, and fructose and the addition of soluble fiber. Gas syndromes may be improved by reduced intake of beans, cabbage, lentils, legumes, apples, grapes, and raisins. Colonic motor dysfunction may be overcome by the gradual addition of combinations of soluble and insoluble fiber containing foods and supplements. The selective use of activated charcoal and simethicone may be helpful. PMID- 2553607 TI - Pseudomonas elastase acts as a virulence factor in burned hosts by Hageman factor dependent activation of the host kinin cascade. AB - Purified Pseudomonas elastase injected subcutaneously into the skin of an Evans blue dye-injected (intravenously) guinea pig caused dye leakage similar to that observed when histamine or bradykinin was injected in the same animal. The histamine-induced dye leakage was ablated in antihistamine-treated guinea pigs, but elastase- and bradykinin-induced dye leakages were not. Local injections of specific inhibitors of the host Hageman factor-dependent bradykinin-generating pathway given immediately prior to elastase injection reduced dye leakage in a dose-related manner. Elastase-related dye release was enhanced when angiotension converting enzyme inhibitor, a substance which prevents host enzymes from breaking down bradykinin, was injected prior to elastase injection. We conclude that Pseudomonas elastase generates bradykinin in the infected host via a Hageman factor-dependent pathway. PMID- 2553608 TI - Pentoxifylline modulation of plasma membrane functions in human polymorphonuclear leukocytes. AB - Pentoxifylline is known to have major effects on cell membrane function in mammalian cells, including human leukocytes. The protective effects of this agent in animal models of infection and inflammation may be due to alterations in phagocyte (neutrophil and macrophage) function. However, the exact mechanism of action of pentoxifylline is unknown. In this study, we evaluated the effect of the drug on several membrane-associated activities in human polymorphonuclear neutrophils and investigated possible mechanisms for the observed changes in neutrophil function. Pentoxifylline inhibited ingestion of microbial particles (Staphylococcus aureus and zymosan); decreased superoxide generation activated by zymosan, formyl-methionyl-leucyl-phenylalanine, and concanavalin A (but not phorbol myristate acetate); and decreased uptake (transport) of adenosine stimulated by formyl-methionyl-leucyl-phenylalanine and zymosan. In contrast, pentoxifylline actually increased clindamycin uptake in zymosan-stimulated polymorphonuclear neutrophils. However, pentoxifylline had no effect on uptake of adenosine or clindamycin in unstimulated neutrophils. In comparison with known inhibitors of nucleoside transport (nitrobenzylthioinosine and dipyridamole), the results suggested that pentoxifylline does not bind to membrane nucleoside transport receptors. At concentrations which inhibit neutrophil function, pentoxifylline activity is not mediated through external membrane nucleoside regulatory sites. Thus, pentoxifylline affects the activation signal chain at a point beyond the membrane receptors. Whatever its precise mechanism of action, pentoxifylline has a striking modulatory effect on cell membrane-associated responses in stimulated leukocytes and may prove useful for control of injurious inflammatory states. PMID- 2553609 TI - Lectinlike interactions of Fusobacterium nucleatum with human neutrophils. AB - Fusobacterium nucleatum expresses lectinlike adherence factors which mediate binding to a variety of human tissue cells. Adherence is selectively inhibited by galactose, lactose, and N-acetyl-D-galactosamine. In this study, adherence of F. nucleatum to human peripheral blood polymorphonuclear neutrophils (PMNs) was investigated. The results indicated that the fusobacteria adhered to live and metabolically inactivated or fixed PMNs. Adherence of F. nucleatum resulted in activation of PMNs as determined by PMN aggregation, membrane depolarization, increased intracellular free Ca2+, superoxide anion production, and lysozyme release. Transmission electron micrographs showed that F. nucleatum was phagocytized by the PMNs. Microbicidal assays indicated that greater than 98% of F. nucleatum organisms were killed by PMNs within 60 min. Adherence to and activation of PMNs by F. nucleatum were inhibited by N-acetyl-D-galactosamine or lactose greater than galactose, whereas equal concentrations of glucose, N-acetyl D-glucosamine, mannose, and fucose had little or no effect on F. nucleatum-PMN interactions. Pretreatment of the fusobacteria with heat (80 degrees C, 20 min) or proteases inhibited adherence to and activation of PMNs, but superoxide production was also stimulated by heated bacteria. The results indicate that interaction of F. nucleatum with PMNs is lectinlike and is probably mediated by fusobacterial proteins which bind to other human tissue cells. Adherence of F. nucleatum to PMNs in the absence of serum opsonins, such as antibodies and complement, may play an important role in PMN recognition and killing of F. nucleatum in the gingival sulcus and in the subsequent release of PMN factors associated with tissue destruction. PMID- 2553610 TI - Purification and properties of a 75-kilodalton major protein, an immunodominant surface antigen, from the oral anaerobe Bacteroides gingivalis. AB - A 75-kilodalton major protein (75K protein) was purified to homogeneity from the cell lysate fraction and the envelope of Bacteroides gingivalis 381. The 75K protein was originally present in the outer membrane or the outermost part of this organism as a large, stable complex with an apparent molecular weight of about 2,000,000. Heating at 80 degrees C and at higher temperatures in the presence of sodium dodecyl sulfate was needed to completely dissociate it to monomers. Amino acid analysis revealed that the 75K protein had about 50% nonpolar amino acids. Various strains of B. gingivalis but not other bacteria, including oral Bacteroides species tested, contained serologically related 75K proteins when tested in Western blotting (immunoblotting) analysis. The abundance and localization of the 75K protein in this organism suggest that it has the potential to participate in the host-parasite interaction in infection. The 75K protein was, indeed, strongly recognized in patients with adult periodontal diseases. Immunoblotting with sera from patients and with rabbit antisera generated by intravenous inoculations of whole B. gingivalis cells revealed that the 75K protein was an immunodominant antigen on the surface of B. gingivalis. PMID- 2553611 TI - Reassessment of the role of splenic leukocyte oxidative activity and macrophage activation in expression of immunity to malaria. AB - The role of splenic leukocyte oxidative activity and macrophage activation in the development of protective immunity was examined during acute Plasmodium chabaudi adami malaria. Splenic leukocyte oxidative activity was compared in infected BALB/c and P/J mice; the latter are known to suffer from defects in macrophage function. Phorbol myristate acetate-stimulated chemiluminescence and superoxide anion production by splenic leukocytes from infected BALB/c mice were found to be increased dramatically, while the response of splenic leukocytes from infected P/J mice was elevated only minimally. Hydrogen peroxide release was slightly increased in splenic leukocytes from infected BALB/c mice but remained essentially unchanged in those from infected P/J mice. Macrophage function was assessed on the basis of measurements of tumoricidal activity. Splenic macrophages from uninfected BALB/c mice displayed significant tumoricidal activity against L929 target cells. As a result of splenomegaly during infection, tumoricidal activity, when calculated on a per-spleen basis, was increased further in infected BALB/c mice. In contrast, the tumoricidal activity of splenic macrophages from P/J mice was minimal, regardless of infection. Despite these differences, both strains of mice developed malarial infections that resolved within 16 days. Thus, while the production of reactive oxygen radicals by splenic leukocytes and the phenomenon of macrophage activation have traditionally been associated with the resolution of malarial infection, this study failed to establish a correlation between these parameters and the development of protective immunity to blood-stage infection with P. chabaudi adami. PMID- 2553612 TI - Molecular epidemiologic analysis of the type A streptococcal exotoxin (erythrogenic toxin) gene (speA) in clinical Streptococcus pyogenes strains. AB - A molecular epidemiology analysis was performed with over 440 clinical isolates of Streptococcus pyogenes obtained from 11 different countries in order to determine the frequency of occurrence of the type A streptococcal exotoxin (erythrogenic toxin) gene (speA) among group A strains. The colony hybridization technique employing a specific internal fragment of the speA gene was used for initial screening, and all positive results were further confirmed by the Southern hybridization technique. Among over 300 general strains obtained from patients with a variety of diseases, except scarlet fever (such as tonsillitis, impetigo, cellulitis, pyoderma, abscess, rheumatic fever, and glomerulonephritis), 15% were found to contain the speA gene. Among a group of 146 strains obtained from individuals described as having scarlet fever, 45% were shown to contain the speA gene. Further analysis of the data indicated that strains with certain M- or T-type surface antigens showed a higher (such as M and T types 1 and 3/13) or lower (such as M2, M12, T4, T5, and T28) tendency to contain the speA gene. No correlation was found between speA content of a strain and the ability to cause a specific disease, although strains possessing the speA gene were more likely to be associated with scarlet fever and rheumatic fever than with other types of disease. PMID- 2553613 TI - A method for allelic replacement that uses the conjugative transposon Tn916: deletion of the emm6.1 allele in Streptococcus pyogenes JRS4. AB - The emm6.1 allele of Streptococcus pyogenes JRS4 was deleted by using the conjugative transposon Tn916. The aphA-3 gene, conferring resistance to kanamycin, was cloned between the sequences flanking the structural gene for the type 6 M protein (emm6.1) and inserted into the BstXI site of Tn916 to generate the chimeric transposon Tn916-5K3. Because the BstXI site lies in a nonessential region of Tn916, the chimeric transposon could transfer by conjugation from Bacillus subtilis into JRS4. In some of the transconjugants, Tn916-5K3 replaced the emm6.1 locus of JRS4 by homologous recombination between the cloned emm6.1 flanking regions and the resident chromosome. One recombinant studied in detail, JRS75, was kanamycin resistant and tetracycline sensitive and lacked immunologically detectable M6 protein. Furthermore, by Southern blot analysis, the DNA region encompassing the emm6.1 structural gene was found to have been replaced by aphA-3. PMID- 2553614 TI - Purification and characterization of two Listeria ivanovii cytolysins, a sphingomyelinase C and a thiol-activated toxin (ivanolysin O). AB - The strong bizonal hemolysis on blood agar and the positive CAMP reaction with Rhodococcus equi denotes the production of two different cytolytic factors by Listeria ivanovii. One was characterized as a thiol-activated (SH) cytolysin of 61 kilodaltons and was termed ivanolysin O (ILO) since data suggested that it is different from listeriolysin O, the SH-cytolysin produced by Listeria monocytogenes. The other is a 27-kilodalton hemolytic sphingomyelinase C that was found to be the cytolytic factor responsible for the halo of incomplete hemolysis synergistically enhanced by R. equi exosubstances. When thiol-disulfide exchange affinity chromatography and gel filtration were applied to the purification of ILO from concentrated L. ivanovii culture supernatants, the copurification of the two cytolysins was observed. This phenomenon seems to be due to the formation of intermolecular disulfide bonds between ILO and the sphingomyelinase, since the latter was found to contain free SH groups, not essential for the activity. These SH groups could react with the single cysteine residue characteristically present in the SH-cytolysins, forming a dimeric cytolytic complex. The purification of ILO was achieved by a further gel filtration with a reducing agent (dithiothreitol) in the eluent. A method for the purification of the sphingomyelinase based on selective sequestration of ILO from the L. ivanovii concentrated culture supernatant by the SH cytolysin target molecule cholesterol and thiol-disulfide affinity chromatography is described. PMID- 2553616 TI - The tachykinins neurokinin A and physalaemin stimulate murine thymocyte proliferation. AB - The tachykinins constitute a family of neuropeptides that are released from sensory neurons, mediating a variety of responses termed neurogenic inflammation. The present study investigates the possibility that tachykinins are also involved in immune-regulatory mechanisms. The mammalian tachykinins neurokinin A (NKA), neurokinin B, neuropeptide K and substance P, as well as the nonmammalian tachykinin physalaemin (PHY) and eledoisin, were analysed in 10-pM to 1.0 microM concentrations for regulatory influences in several lymphocyte proliferation assays. NKA, and to a lesser extent PHY, but none of the other tachykinins tested, displayed a stimulatory action in murine thymocyte cultures, utilised as an interleukin-1 (IL-1) bioassay. The effect was apparent only at a concentration of 0.1 microM or higher. No further stimulatory effect of the tachykinins could be observed in thymocyte cultures already suboptimally stimulated to proliferation by addition of IL-1. The tachykinins had no effect in direct and co mitogenic T and B lymphocyte proliferation assays with rat spleen cells, in a thymocyte growth peptide assay with mouse thymic lymphoblasts or in an interleukin-2 (IL-2) bioassay with IL-2-dependent rat splenoblasts. Our findings indicate that NKA and PHY can act as immune regulators. The results are relevant for the understanding of the interaction between the nervous and the immune system, and are of particular interest in view of the inflammatory actions of both tachykinins and IL-1. PMID- 2553615 TI - Test atmospheres of diisocyanates with special reference to controlled exposure of humans. AB - An all glass apparatus for the generation of air concentrations of 2,4-toluene diisocyanate (2,4-TDI), 2,6-toluene diisocyanate (2,6-TDI) and 1,6-hexamethylene diisocyanate (HDI) was developed. The generation principle was based on gas-phase permeation with permeation membranes of silicon rubber. In an 8 m3 stainless steel test chamber, low and steady TDI-and HDI atmospheres (1-100 micrograms/m3) could be maintained. The diisocyanate concentrations were determined by an HPLC method, using the 9-(N-methylaminomethyl)-anthracene reagent utilizing UV detection. The sum of diisocyanates and their related amines were determined by sampling in 0.4 M hydrochloric acid solution, and analysis by capillary gas chromatography with thermionic specific detection. Related amines were determined by sampling in ethanol - 0.2% KOH and analysis on GC-TSD. A continuous band-tape monitor was used for the determination of diisocyanates. Losses of diisocyanates in the test chamber were evaluated by measuring the TDI and HDI concentrations at the inlet respectively the outlet of the test chamber. At the outlet of the test chamber, ca 25% of the TDI respectively HDI concentrations were recovered. With a male subject in the test chamber ca 15% of the HDI concentration was recovered. The air flow through the test chamber was ca 10 m3/h. The changes in isomeric composition of airborne TDI, at stopped flow conditions, showed that the decay of the 2,4-isomer was faster than of the 2,6-isomer. No trace of the related amine toluene diamine (TDA) was detected in the test chamber, at TDI concentrations ranging from 20 to 50 micrograms/m3. Sampling losses due to sampling connections were evaluated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553617 TI - Inhibin secretion is influenced by Leydig cells: evidence from studies using the cytotoxin ethane dimethane sulphonate (EDS). AB - Adult male rats given a single intraperitoneal injection of the Leydig cell cytotoxin ethane dimethane sulphonate (EDS) show a significant decrease in testosterone from 7 to 14 days, and elevation of serum FSH and LH levels commencing 7 days after treatment, returning to normal at 28 days for LH and 49 days for FSH. A significant rise in serum inhibin levels was seen at day 14 after EDS treatment with levels returning to normal at day 49. In a second series of experiments, silastic implants of testosterone, either 2.5 cm or 22.5 cm in length, were introduced subcutaneously into adult male rats which were treated with EDS 10 days later. Both doses of testosterone suppressed basal LH levels but did not significantly change FSH levels. The rise in FSH and LH levels seen in normal rats after EDS treatment did not occur in either group of testosterone implanted rats. However, serum inhibin levels rose significantly in both groups after EDS treatment, suggesting that the rise in serum inhibin levels was not due to stimulation arising from the increase in FSH levels after EDS treatment. The data suggest that the rise in serum inhibin levels after EDS treatment is linked to destruction of the Leydig cells through mechanisms that require further investigation. PMID- 2553618 TI - Evaluation of the relative importance of endocrine and paracrine factors in control of the levels of inhibin in testicular interstitial fluid. AB - This study aimed to identify the role of endocrine (FSH, LH, testosterone) or paracrine (Leydig or germ cell) factors in control of the secretion of inhibin into testicular interstitial fluid (IF). This was done by measuring inhibin and testosterone levels in IF, and serum gonadotrophin and testosterone levels in adult rats following the destruction of Leydig cells with ethane dimethane sulphonate (EDS), alone or in combination with testosterone ester (TE) supplementation at various doses initiated at various times after EDS treatment. The effect of germ cell loss (induced by local testicular heating) on its own or in combination with the above treatments was also assessed. Treatment with EDS led to major increases in the levels of inhibin in IF and of FSH and LH in serum whilst testosterone levels in IF and serum fell to undetectable levels. Supplementation with TE (1-25 mg) for 21 days from the time of EDS treatment failed to prevent the initial (+3 days) increase in IF levels of inhibin but thereafter suppressed inhibin to control levels or lower and grossly suppressed FSH and LH levels, irrespective of whether the dose of TE administered did (25 or 5 mg) or did not (1 mg) prevent major seminiferous tubule damage. Partial regeneration of Leydig cells and normalization of testosterone levels occurred in rats 21 days after treatment with EDS alone but this failed to normalize inhibin and gonadotrophin levels. When supplementation with TE (25 mg) was initiated at 3, 6 or 9 days after EDS treatment, IF levels of inhibin were normalized within 3 days and maintained thereafter in parallel with suppression of serum FSH and LH to below control levels. Seminiferous tubule damage induced by local testicular heating (43 degrees C for 30 min) led to increased IF levels of inhibin 3 and 14 days later, in parallel with increased serum levels of FSH (but not LH). Suppression of FSH to subnormal levels in heat-exposed rats by TE treatment (25 mg) restored IF inhibin to control levels or below, a change which still occurred when Leydig cells were destroyed by EDS treatment. It is concluded that secretion of inhibin via the base of the Sertoli cell into testicular IF is controlled primarily by FSH, although local factors may play a minor role. These findings have important implications regarding the possible paracrine role(s) of inhibin in IF during puberty and in the normal adult testis. PMID- 2553619 TI - Crystal induced arthropathy in uremia. PMID- 2553620 TI - Glucosaminylmuramyl dipeptide-induced changes in murine macrophage metabolism. AB - The purpose of the study was to investigate influence of glucosaminyl muramyl dipeptide (GMDP) and its derivatives on adenosine deaminase (ADA, CE 3.5.4.4.) and 5'-nucleotidase (5-N, CE 3.1.3.5) activity in murine macrophages in vitro. The intensity of superoxide radicals (O2-) formation by these cells has been also studied. GMDP incubated with macrophages was found to inhibit substantially the activity of 5-N, without affecting the activity of ADA in these cells. The maximal effect on 5-N activity was noted following 24 h of co-culture and was accompanied by a higher intensity of O2- formation. GMDP added in doses ranging from 0.01 to 1 microgram/ml induced a gradual decrease in 5-N activity, with an increase in activity of the O2- -generating system. The GMDP analog with double dipeptide link GM(DP)2 has demonstrated the same activating effect as GMDP. The presence of dipeptide alanyl-D-isoglutamine in the GMDP structure is necessary for realization of the drug activating effect, as N-acetylglucosaminyl-N acetylmuramyl failed to influence macrophage activity. Neither D-D nor L-L isomers of the drug affect the 5-N activity and O2- formation in macrophages. The mechanism of macrophages activation induced by GMDP may include the inhibition of 5-N activity and the stimulation in production of superoxide radicals. PMID- 2553621 TI - Antitumor activity of enkephalin analogues in inhibiting PYB6 tumor growth in mice and immunological effects of methionine enkephalinamide. AB - Recent evidence has implicated enkephalins as immunomodulators. Several studies have reported the regulation of tumor growth by methionine enkephalin (ME). However, there has been little effort to relate the immunological significance of enkephalins to the development of anticancer drugs. The present study had three aims: first, to compare the antitumor activity of the synthetic peptide, D [Ala2]methionine enkephalinamide (MEA), with endogenous enkephalins on PYB6 fibrosarcoma tumor growth; second, to determine whether tumor growth inhibition was mediated by an opiate receptor; and third, to investigate the effects of MEA on selected immune responses. Female B6C3F1 mice were injected i.p. daily for 7 days with 50-4000 micrograms/kg of ME, MEA, leucine enkephalin (LE) or D [Ala2]leucine enkephalinamide (LEA), beginning 1 day after PYB6 inoculation. ME and MEA, but not LE or LEA, decreased the PYB6 growth rate. The dose of 50 micrograms/kg MEA exerted the maximum inhibition of tumor growth (nearly 72% on day 15 post tumor transplantation). MEA was not directly toxic to PYB6 tumor cells, as evaluated by the measurement of DNA synthesis and cellular ATP levels of PYB6 cells exposed to MEA in vitro. No [3H]-etorphine specific bindings were detected on the cell membrane or sonicates of splenic lymphocytes or PYB6 cells. Therefore, the antitumor activity by MEA is likely mediated by an indirect mechanism. Immunological studies indicated that MEA selectively enhanced the lymphoproliferative response to the T-cell mitogen, concanavalin A, but not to the B-cell mitogen, lipopolysaccharide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553622 TI - Entamoeba histolytica depresses chemiluminescence in stimulated human polymorphonuclear leukocytes. AB - Effect of Entamoeba histolytica proteinase/toxin (Ehp/t) on the luminol-dependent chemiluminescence (CL) in stimulated human polymorphonuclear leukocytes (PMN) was studied. The role of superoxide (SO) and hydroxyl (OH) anions in the Ehp/t associated enhancement/inhibition of CL was also studied using specific scavengers and a biological response modifier, muramyldipeptide (MDP). Ehp/t was isolated from axenic trophozoites of the HM-1:IMSS strain of virulent strain of E. histolytica. Proteinase activity was assayed on a synthetic substrate, Z-arg arg-AFC and cytotoxicity was tested on HeLa cell monolayers. PMN isolated from blood were incubated with Ehp/t prior to stimulation by phorbol myristateacetate (PMA, 2 micrograms/ml), serum-treated zymosan (2.5 mg/ml) and glucan (2 mg/ml). CL was monitored in an LKB (Wallac) Luminometer. Ehp/t was found to depress up to 90% of CL induced by PMA, glucan and zymosan. Such a depression was Ehp/t concentration-dependent. A 150 micrograms/ml concentration of Ehp/t, obtained from a 0.015-1.5 mg/ml concentration range tested at different incubation times and temperatures, was used in most of our experiments. Incubation time and temperature optima were 15 min and 37 degrees C, respectively. Ehp/t partially inhibited the CL associated with SO and OH. MDP, in the presence of Ehp/t, enhanced CL response in human PMN to about 67% with reference to normal CL without inhibitor. PMN were confirmed to play a vital role in amebic tissue invasion mechanisms. PMID- 2553623 TI - Effect of arachidonic acid metabolites on CR1 expression by B-lymphocytes. AB - The effect of arachidonic acid metabolites on the expression of the receptor for the C3b/C4b fragment of complement (CR1) by human B-lymphocytes was investigated. Kinetic experiments to determine CR1 expression over time indicated that the maximal receptor number occurred at 2 h, followed by a return to baseline values. Addition of 10(-4) M puromycin to the cells suggested that the increase was due to the expression of an intracellular pool and not de novo synthesis of new receptor molecules. B-lymphocytes were incubated with arachidonic acid, 15 hydroxyeicosatetraenoic acid, leukotrienes B4 or C4 or prostaglandin E2 (PGE2). The quantity of membrane antigenic binding sites was determined before and after incubation. The lipoxygenase metabolites did not alter CR1 numbers. In contrast, PGE2 significantly decreased (P less than 0.05) the quantity of CR1 expressed. In kinetic experiments, PGE2 blocked the maximal expression of CR1 seen at 2 h, indicating that it prevents the appearance of an intracellular pool of receptor. These results show that CR1 number on B-lymphocytes can be altered by at least one arachidonic acid metabolite. This may offer a partial explanation for the inhibitory effects of PGE2 on B-cell proliferation and immunoglobulin secretion since CR1 is implicated in B-lymphocyte differentiation and specific antibody response. PMID- 2553624 TI - Chemotactic response of rat macrophages is enhanced by two diastereoisomers of LTB4. AB - We demonstrated that rat peritoneal cells synthesize, and release into the culture medium, substances that elicit a chemotactic responsiveness of rat macrophages. These substances were identified by HPLC analysis as two diastereoisomers of LTB4, precisely delta-6-trans-LTB4 and delta-6-epi-trans LTB4. Peritoneal cells release, also, other lipoxygenase metabolites such as LTD4 and LTB4, which were shown not to be active in eliciting chemotaxis of rat macrophages, in agreement with data of other authors. Quantitative assays for the measurement of PMNs or macrophage chemotaxis were carried out in Boyden chambers. Release of leukotrienes by rat peritoneal cells into the culture medium was strongly enhanced upon stimulation with calcium ionophore A23187, an agent known to increase cytoplasmic Ca2+ concentration, thus leading to the activation of Ca2+-dependent phospholipase and a consequent increase in the amount of endogenous free arachidonic acid available for biotransformation. The action of several inhibitors of arachidonate metabolism at concentrations more selective for the lipoxygenase than the cycloxygenase pathway, was also studied. alpha Tocopherol and ASA were shown to be the most selective in inhibiting the synthesis of both the LTB4 diasteroisomers active as enhancers of rat macrophage chemotaxis. PMID- 2553625 TI - Stimulation by a bacterial extract (Broncho-Vaxom) of the metabolic and functional activities of murine macrophages. AB - Peritoneal and bone marrow-derived macrophages of the C57BL/6 and DBA/2 mouse strains were exposed in vitro to increasing concentrations of the bacterial lysate Broncho-Vaxom (BV), in the presence or absence of macrophage-activating factor (MAF)-rich media. Two metabolic pathways and two functional activities of the macrophages were studied. First, oxidative metabolism was found to increase sharply in macrophages incubated with BV, as measured by the catabolism of glucose via the hexose monophosphate shunt pathway, and by the production of the superoxide anion (O2-). Both effects were further increased by co-stimulation of macrophages with MAF. Second, exposure to BV together with MAF (or with recombinant murine interferon-gamma) led to acquisition by macrophages of the capacity to destroy the intracellular parasite Leishmania enriettii; such activated macrophages were also lytic towards P815 mastocytoma indicator target cells. These cytotoxic properties failed to develop in the absence of MAF. The BV dependent increase in metabolic and functional activities was of the same magnitude as that induced by incubation of macrophages with 10 ng/ml of bacterial lipopolysaccharide (LPS). Residual contamination of BV by endotoxin was however much lower. In addition, polymyxin B, a LPS inhibitor, blocked the effect of LPS without significantly affecting macrophage stimulation by BV. These experiments indicate that BV can markedly stimulate macrophage metabolic and functional parameters that are important for host defense against pathogens and tumors. PMID- 2553626 TI - Dibutyryl guanosine cyclic monophosphate causes proliferation of interleukin-2 dependent T-cells in the presence of suboptimal levels of interleukin-2. AB - The effect of dibutyryl guanosine cyclic monophosphate (db-cGMP) on proliferation of an interleukin-2 (IL-2)-dependent T-cell line was investigated. Dibutyryl cGMP added to HT-2 cell cultures without IL-2 or in the presence of an optimal IL-2 concentration (60 U/ml) had no effect on proliferation. The same amount of db cGMP (10(-5)M) added to cultures in the presence of suboptimal amounts of IL-2 (3 6 U/ml) produced significant increases in proliferation. In contrast, db-cAMP caused substantial inhibition of HT-2 cells cultured with IL-2. Similar effects were observed using human T-cell blasts produced after 72 h stimulation with phytohemagglutin. These results suggest that cyclic GMP may play a role in the cellular response of IL-2 responsive cells to IL-2. PMID- 2553627 TI - Studies on the oncogenic potential of Epstein-Barr-virus (EBV)-infected B cells in AIDS-related disorders. AB - Spontaneous lymphoblastoid cell lines (LCLs) were established from the peripheral blood of 10 human immunodeficiency virus (HIV)-seropositive patients in order to investigate whether or not progression of the cells towards a malignant state could be traced. The LCLs studied displayed no differences in their surface phenotype, karyotype, and tumorigenicity in nude mice as compared with a wide panel of control LCLs. Furthermore, no c-myc rearrangement could be detected in any of the LCLs. However, 4 of the 10 LCLs derived from HIV-seropositive patients formed colonies in agar with a cloning efficiency of 0.1-0.9%. This percentage was much lower than that of a control neoplastic B cell line (50%), but consistently higher than that observed for a battery of spontaneous LCLs. The cells of a number of sublines that were derived from the agar colonies expressed new activation markers (CD10 and Bac-1) but did not induce tumors in nude mice or display chromosomal abnormalities. These sublines might comprise cells that have progressed towards a more markedly transformed state. PMID- 2553628 TI - Hilus cell tumor of the ovary. A clinicopathological analysis of 12 Reinke crystal-positive and nine crystal-negative cases. AB - The clinical and pathological features of 12 Reinke crystal-positive and nine probable (crystal-negative) hilus cell tumors of the ovary were reviewed. The patients' ages ranged from 32 to 82 years. The initial manifestation was androgenic in 62% of the cases. The levels of testosterone were high (200-2,400 ng/dl) even though the tumors had a mean diameter of only 2.1 cm. All the tumors were centered in the hilus of the ovary, extending for varying distances into the ovarian stroma. They were composed of polyhedral to rounded cells with generally abundant eosinophilic cytoplasm and often showed a perivascular aggregation of nuclei with avascular pooling of cytoplasm or hyalinization of stroma. Fibrinoid changes in the vessels of the tumors were frequently noted. None of the 10 patients for whom follow-up data of 1-17 years were available died of tumor; virilizing symptoms regressed but did not completely disappear in five of the 10 patients after the removal of the tumor. PMID- 2553629 TI - Human papillomavirus messenger RNA expression in adenocarcinoma in situ of the uterine cervix. AB - Over recent years, the association between human papillomavirus (HPV) and squamous cell carcinoma of the cervix and its precursors has been well established, largely as a result of advances in the techniques of molecular biology. A similar association between HPV and invasive adenocarcinoma of the cervix has also been recently demonstrated; however, little work has been published on the relationship between the precursor lesion, adenocarcinoma in situ (ACIS), and HPV. We have therefore undertaken an in situ hybridization study of 22 cases of known cervical ACIS using probes for HPV messenger RNA expression. Seventeen of these had residual ACIS in the blocks studied, 15 of which (88.6%) expressed HPV messenger RNA. Five cases were positive for HPV 16 and 10 cases for HPV 18. Early invasive adenocarcinoma was present with ACIS in three cases and one case had early invasion plus cervical intraepithelial neoplasia (CIN III). These invasive lesions showed a similar type and degree of HPV expression as the in situ component. One section had only residual CIN III, which was positive for HPV type 18. Four cases had only minor glandular atypias (less than ACIS) in the studied sections, and all four of these were negative for HPV expression. None of the normal endocervical epithelia in any of the sections were positive by this technique. In summary, the finding of HPV messenger RNA expression in nearly 90% of cervical ACIS supports a possible role for these viruses in the pathogenesis of glandular neoplasms of the uterine cervix. PMID- 2553630 TI - A study of nucleolar organizer regions in cervical intraepithelial neoplasia and human papillomavirus infection. AB - A silver colloid technique for nucleolar organizer region-associated proteins (AgNORs) was applied to sections of cervix that comprised cervical intraepithelial neoplasia (CIN) grades I to III, koilocytosis indicative of human papillomavirus infection, squamous metaplasia, and basal cell hyperplasia. The AgNOR count was compared both within and between specimens. Proliferative activity was investigated by flow cytometric analysis. Significant differences in AgNOR numbers were found between normal internal controls and the lesions investigated. A correlation was found between the proliferative index (%S + G2) determined by flow cytometry and the number of AgNORs in cases of CIN. PMID- 2553631 TI - Practical approach to cervical human papillomavirus-related intraepithelial lesions. AB - Members of the Editorial Board who have been active in the study of female genital HPV-related lesions have responded to a series of questions related to the histopathologic diagnosis of such lesions in the uterine cervix, the performance of auxiliary studies, therapeutic choices, and the correlation of cytologic and tissue diagnoses. PMID- 2553632 TI - Conformational analysis of enkephalin analogs containing a disulfide bond. Models for delta- and mu-receptor opioid agonists. AB - Conformational analysis of the cyclic opioids H-Tyr-D-Pen-Gly-Phe-D-Pen-OH (DPDPE) and H-Tyr-D-Cys-Gly-Phe-D-Cys-OH (DCDCE) have been performed using the AMBER program. DPDPE is considerably more selective for delta-receptors than DCDCE. Using the RNGCFM program, a large number of ways were found to close the 14-membered disulfide-containing ring structure. However, intramolecular hydrogen bonds were only possible in gamma-turn and inverse gamma-turn conformations centered on the glycine residue which were associated with opposite chiralities of the disulfide bond. With the cyclic part of the molecules in either a gamma turn or inverse gamma-turn, a systematic conformational analysis was performed on the tyrosine and phenylalanine sidechains. This showed that conformers with the tyrosine and phenylalanine phenyl rings in the vicinity of the disulfide bond were preferred due to attractive van der Waals forces. For DPDPE, however, this was only possible with a positive dihedral angle for the disulfide bond due to the presence of the beta-carbon methyls of Pen2. In contrast, these preferred conformers were possible with both chiralities of the disulfide bond in DCDCE. Conformational entropies and free energies were computed from the translational, rotational, and vibrational energy levels available to each conformer. The conformational entropies were found to vary significantly and to result in a re ordering of the lowest energy minima. Based on these conformational differences in DPDPE and DCDCE and their differing pharmacological selectivities, tentative conformational preferences for delta- and mu-receptor opioid peptides are proposed. PMID- 2553633 TI - Melanotic neuroectodermal tumor of infancy: report of two cases. AB - Melanotic neuroectodermal tumor of childhood is a rare lesion, but its presentation appears to be quite characteristic and a clinical diagnosis may be made with some confidence. Although this is an extremely rapidly growing, aggressive tumor, local excision appears to be adequate for cure. PMID- 2553634 TI - Ipsilateral responses induced by factors present in left and right hemispheres. AB - The extract of left (LVC) and right (RVC) turtle visual cortex locally applied to the surface of turtle visual cortex inhibited the orthodromic evoked potentials (EP; fast negative component N1). The inhibition was partially (70-90%) prevented by opiate antagonist naloxone. LVC-extract proved to be a more potent inhibitor of the left cortex EP, whereas RVC-extract was slightly more effective when applied to the right cortex. The extracts of the left and right hemispheres of rat brain, but not the extract of the whole brain, applied subarachnoidally into the caudal portion of a transected spinal cord at the T7-T9 level, induce postural asymmetry of hind limbs in rats. The extract of the left hemisphere predominantly induce flexion of the left leg, and the extract of the right hemisphere induce flexion of the right leg. The factors of the right brain hemisphere able to induce postural asymmetry (PAFs) have been studied in detail. PAFs are thermostable compounds inactivated by trypsin and papain. Naloxone significantly reduces the number of asymmetries. HPLC analysis of the rat and pig right hemisphere extract has revealed two PAFs which differ from enkephalins, endorphins and dynorphin. The activity of PAFs in rat pituitary is significantly lower than in the brain. These findings suggest the existence of side-specific mechanism for selective neurohormonal regulation of neuronal activity and other processes in the left and right halves of the brain which involve lateralized factors of turtle visual cortex and PAFs. PMID- 2553635 TI - ACTH and its relationship to the cataplectic syndrome. PMID- 2553636 TI - Thymomodulin increases the depressed production of superoxide anion by alveolar macrophages in patients with chronic bronchitis. AB - Thymomodulin is an immunomodulating agent which is derived from calf thymus by partial acid lysis. It promotes T-cell maturation, enhances antibody synthesis and improves the phagocytic response of neutrophils. Clinical trials have revealed the effectiveness of this thymic derivative in the prevention of recurrent respiratory infections (RRI) in children and in adults; 11 patients (8 males and 3 females; age range 18-76 years) with chronic bronchitis dominated by recurrent respiratory infections were studied. They were treated orally for 6 months during the winter season with 120 mg/day of thymomodulin. All the subjects were asked to keep a diary recording the intensity of their symptoms, the number of working days lost (days of illness) and the use of antibiotic and/or mucolytic drugs. At the beginning and at the end of the trial each patient was subjected to a control with a flexible fibreoptic bronchoscope with bronchoalveolar lavage to evaluate the phagocytic response of alveolar macrophages. At the end of therapy a significant improvement of the clinical status, evaluated by the above-mentioned parameters, of the bronchial mucosa aspect and an increase in alveolar macrophage superoxide production was noticed (from 0.1 +/- 0.09 and 0.8 +/- 0.5 nmol to 1.6 +/- 0.8 and 4.1 +/- 2.2 nmol with PMA or zymosan particles respectively; p less than 0.001). During thymomodulin treatment no side-effects were recorded. PMID- 2553637 TI - [Endocrine pancreatic tumor with Cushing syndrome and hypergastrinemia]. PMID- 2553638 TI - A Na/H exchange mechanism in apical membrane vesicles of the retinal pigment epithelium. AB - The retinal pigment epithelium (RPE) interposed between the vascular system of the choroid and the neural retina performs a variety of functions essential for vision. In order to further elucidate the transport functions of the RPE, apical membranes were isolated from the RPE of the dogfish (Squalus acanthias) by differential precipitation with calcium. Na-K-ATPase, an apical marker enzyme in this tissue, was enriched 15-fold in the final membrane fraction. About 50% of the membranes form right-side-out vesicles in which the membrane has retained its in vivo orientation. Sodium uptake into these vesicles as determined by a rapid filtration method was stimulated 37% by the presence of a proton gradient across the membrane (pHi = 6.1, pHo = 8.1). The stimulation was also observed in membrane vesicles "short-circuited" with valinomycin and K. The pH gradient dependent sodium uptake but not the uptake in the absence of a pH gradient was completely inhibited by 5 X 10(-4) M amiloride, and 56% inhibition was found at 10(-5) M amiloride. The uptake of 22Na was also strongly decreased in the presence of nonradioactively labelled sodium and lithium; potassium was without effect. pH gradient dependence, amiloride sensitivity, saturability and cation specificity of the sodium flux indicate the presence of a Na/H exchanger in the apical membrane of the retinal pigment epithelium. The presence of the Na/H exchange process might have important implications for the control of pH in the subretinal space, optimum intracellular pH of the RPE and the triggering of other functions of the RPE. PMID- 2553639 TI - Atrial natriuretic peptides. Effects on intraocular pressure, cGMP, and aqueous flow. AB - We report a significant decrease in intraocular pressure after intravitreal injection of atrial natriuretic peptide (AP). There is also an increase in cGMP in iris-ciliary body preparations after exposure to physiologic concentrations of AP in vitro. We present evidence that after intravitreal AP administration there is a decrease in aqueous humor flow associated with the decrease in intraocular pressure. The changes in aqueous humor flow were estimated by automated fluorophotometry and aqueous humor ascorbate concentrations. PMID- 2553640 TI - The potential malignancy of eccrine spiradenoma. AB - Eccrine spiradenoma is a rare, usually benign tumor, originating from cutaneous sweat glands. Its clinical and histologic diagnosis can be difficult at times, especially when malignant features are present. Proper management requires radical surgical excision. Two cases of spiradenoma are reported, one of which highly malignant with diffuse pulmonary metastases leading to the patient's death in spite of repeated radical surgery. PMID- 2553641 TI - Increased plasma beta-endorphin levels in hereditary angioedema. AB - We measured beta-endorphin (BE) and beta-lipotropin (BLPH) plasma concentrations (by means of an HPLC-RIA coupled method) during attacks as well as during symptom free periods in a group of 28 patients with immunochemical (21) or functional (7) Cl inhibitor deficiency. Thirteen patients suffering from chronic urticaria served as controls. Three orders of considerations prompted us to initiate the present study: the clinical relationship between stress and the onset of acute episodes, the possible effects of repeated stressful situations, as are the attacks themselves, on the patients' neuroendocrine system and the well-known existence of close links between the immune system and endogenous opioids. The results show that plasma BE (and, to a lesser extent, BLPH) is dramatically increased during the attacks. In symptom-free periods many patients show very high BE concentrations, often in the presence of slightly elevated concentrations of BLPH and of ACTH. These observations suggest that patients with hereditary angioneurotic edema show a modified pro-opiomelanocortin-synthesizing cell activity that can result in a massive release of BE from the readily disposable pool present in the pituitary and/or an increase in the turnover of the peptide as evaluated by the BLPH/BE ratio. PMID- 2553642 TI - The effect of intravenous immunoglobulin on the in vitro function of human neutrophils. AB - Three commercially available preparations of human immunoglobulin for intravenous use (IVIgG), namely Gamimune N. Sandoglobulin and Intraglobin F, were tested for their ability to modulate human neutrophil function in vitro. IVIgG consistently stimulated the neutrophil respiratory burst at concentrations of 0.5 to 1 mg/ml, concentrations readily achieved in vivo by moderate-dose therapy. Superoxide (O2 ) release was increased by 3.5-4.5 nmol per 5 x 10(5) cells at these concentrations of IVIgG, and H2O2 production increased in a dose-dependent fashion up to 8 mg/ml IVIgG. Luminol-dependent chemiluminescence (CL) was also directly stimulated by IVIgG. In addition, the effects of both soluble and particulate stimulators (N-formyl-methionyl-leucyl-phenylalanine, phorbol myristate acetate and opsonized zymosan) on the neutrophil respiratory burst were enhanced by IVIgG. In a filter assay of neutrophil migration, using a modified Boyden chamber, no consistent effect on neutrophil locomotion or chemotaxis could be demonstrated. The effect of IVIgG on neutrophil metabolism may contribute to its beneficial therapeutic effect in severe, life-threatening infections. PMID- 2553643 TI - Therapeutic use of anti-Rh (D) immunoglobulin in thrombocytopenia. AB - Anti-Rh (D) immunoglobulin (RhoGAM, Johnson and Johnson, Diagnostics) has been used for idiopathic thrombocytopenic purpura (ITP)1. This report explores its further use in thrombocytopenia of various etiologies. PMID- 2553644 TI - [Disseminated herpes zoster in chronic lymphatic leukemia]. AB - A 73-year-old woman suffering from chronic lymphatic leukemia presented a rare variant of zoster with disseminated eruptions. Ultrastructural negative staining revealed viruses of the herpes group. Herpes simplex was excluded by means of fluorescence-labelled monoclonal antibodies (HSV-1/HSV-2 direct specimen identification/typing test). It is proposed that the conventional classification of zoster into a segmental and a generalized variant should be supplemented. PMID- 2553645 TI - Mutation rates in hybrids between sibling species of Drosophila. AB - It has recently been suggested that sterility or inviability in species hybrids might result from the movement of transposable elements. Because such movement is often detectable by an increased mutation rate, I studied the effect of interspecific hybridization in the Drosophila melanogaster group on the mutation rate of X-linked visibles. This rate did not differ between hybrids and intraspecific controls. This was also true for the germ-line excision rate of a transposable element, although the rate of somatic excision was two to sixfold higher in hybrids than in pure species. Combined with previous work, these results do not support a role for transposable elements in speciation. PMID- 2553647 TI - A genetically engineered AIDS protein. PMID- 2553646 TI - [Surgery of pleomorphic adenoma of the parotid gland]. AB - In literature opinion is divided, whether for treatment of pleomorphic adenoma in the parotid gland partial parotidectomy is adequate or complete parotidectomy is necessary. In a retrospective analysis of 123 complete parotidectomies in pleomorphic adenomas the macroscopic tumor extension was compared with the microscopic findings. Additionally 35 pleomorphic adenomas were examined in histological serial sections. The retrospective analysis showed in 6%, the serial sections in 11% microscopic tumor in the macroscopic tumorfree inner lobe. A clinical follow-up examination of 85 patients operated by complete parotidectomy because pleomorphic adenoma resulted in a relapse-quota of 3.5%. The function of the facial nerve was in most cases normal, in 8.2% a weakness of the mandibular branch was found. These results indicate that the risk to leave tumor can be reduced evidently by a complete parotidectomy. PMID- 2553648 TI - The treatment of advanced juvenile nasopharyngeal angiofibroma. AB - Fifteen patients with juvenile nasopharyngeal angiofibroma (JNA) were treated in the Department of Radiation Oncology, Baylor College of Medicine between 1973 and 1986. All patients underwent radiographic evaluation including CT scanning, selective digital subtraction angiography, tomograms, or MRI. Patients referred for definitive irradiation exhibited extensive tumor involvement. Eleven of 15 patients had middle cranial fossa involvement; cavernous sinus extension was observed in six patients. Ten patients were treated with primary radiation therapy; five patients had surgical resection initially and were referred for radiation therapy upon local recurrence. Follow-up ranges from 1 1/2-13 years. Four of the 5 patients who received 3200 cGy in 200 cGy fractions demonstrated tumor recurrence within 2 years after irradiation. All recurrences were ultimately controlled by either further irradiation and/or resection. No tumor recurrence was encountered among the patients treated at the higher tumor doses (36-46 Gy). No severe complications have been observed. Radiation therapy utilizing carefully tailored fields is an appropriate therapeutic approach to patients with extensive disease or intracranial extension. A total dose of greater than 40 Gy may allow improved local control for advanced lesions. PMID- 2553649 TI - Changes in motor nerve terminals during bupivacaine-induced postsynaptic deprivation. AB - The local anaesthetic agent bupivacaine induces a rapid degeneration of muscle cells when injected in the sternocleidomastoid muscle of the adult rat with no damage to intramuscular nerves. Nevertheless, silver impregnation and electron microscopy reveal a sequence of retraction and sprouting of the motor nerve endings during the period of the bupivacaine-induced muscle cell deprivation by necrosis. These morphological changes seem to be qualitatively similar to the retraction and growth phenomena described as forming part of the remodelling process affecting normal untreated neuromuscular junctions of the adult amphibia and mammals. The results are discussed in the context of the regulatory mechanisms governing the maintenance of motor nerve terminals. PMID- 2553650 TI - IV. Acyl-CoA: 6-APA acyltransferase of Penicillium chrysogenum: studies on substrate specificity using phenylacetyl-CoA variants. AB - Two different penicillins (p- and m-methylbenzylpenicillin) were obtained "in vitro" by direct enzymatic synthesis, using homogeneously pure acyl-CoA: 6 aminopenicillanic acid (6-APA) acyltransferase from Penicillium chrysogenum, 6 APA and p- or m-tolylacetyl-CoA. The Km for these substrates were 6 and 15 mM, respectively, indicating that the affinity of the enzyme for these two molecules is much lower that shown by phenylacetyl-CoA (0.55 mM). Furthermore, acyltransferase does not recognize o-tolylacetyl-CoA as a substrate suggesting that the position of the methyl group on the aromatic moiety may have a very important role in the formation of the enzyme-substrate complex. PMID- 2553651 TI - Nutrition of goats as influenced by thinning and clearing of deciduous woodland in northeastern Brazil. AB - Nutritive composition of diets and forage intake of goats utilizing pastures of cleared, two levels of thinned (25% and 55% canopy cover) and undisturbed (control) tropical woodland were compared on a seasonal basis. Esophageally fistulated goats were used to collect dietary samples for chemical analysis. Total fecal collections were used to estimate organic matter intake (OMI) and digestible energy intake (DEI). Forage availability during the wet season was more than threefold higher (P less than .1) on treated than on control pastures. Although available forage was similar (P greater than .1) for all treatments by the middle of the dry season, available forage on the control pastures was limited mostly to dry tree leaves. Goats selected diets 4% higher (P less than .1) in neutral detergent fiber on the control pastures than on the treated pastures; otherwise, diets selected by the goats on the various pastures were similar in terms of crude protein, lignin and in vitro organic matter digestibility. However, OMI and DEI were 23 and 31% higher (P less than .1), respectively, on the treated pastures than on the control pastures. There were no differences among the cleared and thinned treatments. Excessively wet conditions during the rainy season adversely affected goat performance. High populations of biting insects and wet field conditions reduced foraging time and diet selection. The generally higher levels of available forage on the cleared and thinned pastures did not result in the goats' selecting diets higher in nutritive value on the treated pastures than on the control pastures. Instead, the increased forage availability for the treated pastures allowed the goats to attain higher intake of forage. PMID- 2553652 TI - Effects of dietary protein level and clinoptilolite on the weight gain and liver mineral response of growing lambs to copper supplementation. AB - Growing male Synthetic I (1/2 Finnish Landrace x 1/4 Dorset x 1/4 Rambouillet) lambs were used in two experiments (64 lambs in Exp. 1 and 63 in Exp. 2) to test the hypothesis that dietary CP level (9 or 14% of diet as fed) and(or) clinoptilolite (clino; 0 or 2% of diet) affects growth and tissue mineral concentrations of growing lambs fed supplemental Cu. Lambs were individually fed their respective diets ad libitum and killed after 12 wk (Exp. 1) or 16 wk (Exp. 2) to obtain carcass measurements, organ weights and liver mineral concentrations. In Exp. 1, 20 ppm added Cu (as CuSO4.5H2O) increased mortality and depressed BW gain (P less than .01) and daily feed intake (P less than .05) in the presence or absence of clino and at both levels of CP. Liver Cu concentration was greater (P less than .01) in lambs fed added Cu than in those not fed Cu (408 ppm vs 110 ppm, respectively). Neither CP nor clino affected liver Cu concentration. Clinoptilolite increased daily gain of lambs fed high CP but not low CP (P less than .01). In Exp. 2, clino in the diet had no effect on daily gain or daily feed, but 20 ppm Cu addition depressed daily gain (P less than .01) and gain/feed (P less than .07). Organ weights and levels of trace elements other than Cu in the liver generally were not affected by diet in either experiment. It is concluded that high dietary CP or 2% dietary clino did not protect against toxic signs of Cu when Cu was added to the basal diet (10 ppm Cu) at 10 or 20 ppm. PMID- 2553653 TI - Relation between Aeromonas and faecal coliforms in fresh waters. AB - A possible correlation between the presence of mesophilic aeromonads and the number of faecal coliforms present in three fresh-water habitats subject to differing levels of faecal pollution was investigated. Concentration of Aeromonas spp. between 10(2)-10(9) cfu/100 ml and faecal coliforms of between 9-10(7) cfu/100 ml were found in the waters. In water free from faecal pollution there was no correlation but in polluted waters there was a significant relationship between the numbers of aeromonads, faecal coliform and the concentration of organic matter measured by biological oxygen demand. PMID- 2553654 TI - Itraconazole and fluconazole: new drugs for deep fungal infection. PMID- 2553655 TI - The effect of pentamidine salts on the respiratory burst of human neutrophilic granulocytes. AB - Therapeutic concentrations (0.3-1.5 mg/l) of pentamidine isethionate and pentamidine mesylate, obtained after parenteral administration of either drug, did not affect oxygen consumption in the stimulated neutrophilic granulocyte. At concentrations of 0.7, 1.1 and 1.5 mg/l, superoxide production, hydrogen peroxide production, myeloperoxidase (MPO)-mediated iodination and hexose monophosphate shunt activity were suppressed relative to untreated cells (P less than 0.001 in each case). The depression in each activity was dose-related. There was no significant difference between the drugs with regard to these impairments in neutrophilic granulocyte function. This lowered respiratory burst activity, which would lead to a depression of MPO-dependent and MPO-independent processes in stimulated neutrophilic granulocytes, may be due to drug induced dysfunction of NADPH-oxidase. PMID- 2553656 TI - Susceptibility of anaerobic bacteria to meropenem. AB - The activity of meropenem was determined against 350 strains of anaerobic bacteria by an agar dilution method. It was compared with those of piperacillin, cefoxitin, imipenem, clindamycin, metronidazole and chloramphenicol. Meropenem and imipenem were the most active agents tested. On the basis of these results, meropenem appears to be a promising antimicrobial agent for anaerobic infections and warrants further clinical investigation. PMID- 2553657 TI - Comparative activity of meropenem against Pseudomonas aeruginosa strains with well-characterized resistance mechanisms. AB - Four major mechanisms cause resistance to beta-lactams in Pseudomonas aeruginosa: (i) cell wall impermeability gives broad-spectrum intrinsic resistance to all beta-lactams except imipenem, (i) loss of D-group outer membrane proteins correlates with narrow spectrum imipenem resistance, (iii) plasmid mediated beta lactamases compromise antipseudomonal penicillins, cefoperazone and cefsulodin, and (iv) chromosomal beta-lactamase hyper-production compromises most beta lactams except carbenicillin and imipenem. Meropenem was tested in vitro against P. aeruginosa isolates, mutants and transconjugants with these mechanisms. Meropenem had impaired activity (MIC 1-2 mg/l compared to 0.25 mg/l for sensitive isolates) for organisms with broad-spectrum intrinsic resistance. MICs of meropenem also were elevated (to 1-2 mg/l) for mutants with D2-protein-deficiency associated imipenem resistance. Most plasmids encoding TEM, OXA or PSE beta lactamases did not increase the MIC (0.12 mg/l) of meropenem for P. aeruginosa PU21. Decreased susceptibility (MIC 4 mg/l), however, was observed when plasmids coding the uncommon NPS-1, PSE-2 and OXA-3 enzymes were present in this strain. MICs of meropenem remained identical for chromosomal beta-lactamase-inducible P. aeruginosa strains and their enzyme-derepressed and basal mutants, indicating that the chromosomal beta-lactamase could not protect against the new carbapenem, regardless of its mode of expression. PMID- 2553658 TI - Nutritional requirements of papillomavirus-transformed mouse cells and an uninfected parent line in serum-free culture. AB - A serum-free culture system was used to compare the nutritional requirements of mouse mammary cells transformed by bovine papillomavirus type 1 (ID13 cells) and the uninfected parent line (C127 cells). The serum-free, chemically defined medium used for this study was an MCDB 151-based medium (MCDB 151+S+I), supplemented with epidermal growth factor, transferrin, hydrocortisone, ethanolamine, phosphoethanolamine, retinoic acid, trace metals, and insulin. Proliferation of either cell type in serum-free culture required the addition of 250 micrograms/ml of insulin. ID13 cells have a doubling time of greater than 96 h in MCDB 151+S+I, whereas C127 cells have a doubling time of 60 h. This is in sharp contrast to the growth characteristics of the two cell types in 10% fetal bovine serum, where doubling times for the ID13 and C127 cells are 24 and 30 h, respectively. Culture of the cells in a serum-free medium has therefore revealed that the papillomavirus-transformed cells have more stringent growth requirement than the uninfected parent line. PMID- 2553660 TI - Chemical characterization and bioavailability of phosphorus in water-insoluble fractions of three mono-ammonium phosphate fertilizers. AB - Elemental impurities in mono-ammonium phosphate (MAP) fertilizers, such as Fe, Al, Ca, Mg, and F, result in compounds other than NH4H2PO4. The phosphorus availability of the impurity compounds was determined in 3 commercial MAP fertilizers produced from North Carolina, Florida, and Idaho phosphate rocks. Soluble compounds, including NH4H2PO4, were washed out of the fertilizers, and the water-insoluble fraction was collected. The North Carolina, Florida, and Idaho MAP fertilizers contained 13, 16, and 17% water-insoluble fractions, respectively. Availability of phosphorus in each MAP fertilizer and water insoluble fraction was determined in a greenhouse pot study using sorghum sudangrass (Sorghum bicolor) as the test crop. There were no differences in dry matter weights or phosphorus uptake between reagent grade MAP and each MAP fertilizer. Lower dry matter weights and phosphorus uptake were produced from the water-insoluble North Carolina, Florida, and Idaho fractions (NC greater than FL greater than ID). Chemical analysis and characterization with optical microscopy, infrared spectroscopy, and X-ray diffractometry indicated 77.3, 73.8, and 73.6% NH4H2PO4 in the North Carolina, Florida, and Idaho MAP fertilizers, respectively. MgAl(NH4)2H(PO4)2F2, AlNH4HPO4F2, and FeNH4(HPO4)2 were identified as water insoluble phosphorus compounds. There was a significant negative correlation between the percent phosphorus present as MgAl(NH4)2H(PO4)2F2 and the phosphorus availability. Increased crystallinity of the various water-insoluble compounds also contributed to reduced phosphorus availability. PMID- 2553659 TI - Isolation of fetal mouse motor neurons on discontinuous Percoll density gradients. AB - The spinal cords of fetal NIH:CR mice, gestational age Day 12 to 14, were dissected free of meninges and dorsal root ganglia, chemically dissociated, and layered onto discontinuous Percoll gradients at densities 1.040, 1.050, and 1.060 g/ml. After centrifugation (800 Xg for 15 min at 4 degrees C), three morphologically, biochemically, and immunohistologically distinct cell populations were collected from the gradient interfaces. The first interface, located at a density of 1.040 g/ml, was choline acetyltransferase enriched (0.86 +/- 0.08) compared to the second and third fractions (0.42 +/- 0.01 and 0 pmol acetylcholine synthesized/microgram protein, respectively). When simultaneously cultured with fetal mouse cardiac muscle on a gelatin-polylysine-laminin substrate in serum-free medium, these cells developed the characteristics of motor neurons. PMID- 2553661 TI - Construction, expression, and localization of a CycA::PhoA fusion protein in Rhodobacter sphaeroides and Escherichia coli. AB - We demonstrated the utility of Escherichia coli alkaline phosphatase, encoded by phoA, as a reporter molecule for genetic fusions in Rhodobacter sphaeroides. A portion of the R. sphaeroides cycA gene was fused to phoA, yielding a fusion protein comprising the putative signal sequence and first 10 amino acids of the cytochrome c2 apoprotein joined to the sixth amino acid of alkaline phosphatase. The fusion protein was efficiently transported to the periplasm of R. sphaeroides as determined by enzyme activity, Western immunoblot analysis, and immunogold electron microscopy. We also documented the ability of an R. sphaeroides mutant, RS104, with gross defects in photosynthetic membrane morphology to efficiently recognize and translocate the fusion protein to the periplasmic compartment. The inclusion of 500 base pairs of R. sphaeroides DNA in cis to the cycA structural gene resulted in a 2.5-fold increase in alkaline phosphatase activity in photosynthetically grown cells compared with the activity in aerobically grown cells, demonstrating that the fusion protein is regulated in a manner similar to that of cytochrome c2 regulation. We also constructed two pUC19-based plasmids suitable for the construction of translational fusions to phoA. In these plasmids, translational fusions of phoA to the gene under consideration can be made in all three reading frames, thus facilitating construction and expression of fusion protein systems utilizing phoA. PMID- 2553662 TI - Physical and genetic mapping of the Rhodobacter sphaeroides 2.4.1 genome: genome size, fragment identification, and gene localization. AB - Four restriction endonucleases, AseI (5'-ATTAAT), SpeI (5'-ACTAGT), DraI (5' TTTAAA), and SnaBI (5'-TACGTA), generated DNA fragments of suitable size distributions for mapping the genome of Rhodobacter sphaeroides by transverse alternating field electrophoresis. AseI produced 17 fragments, ranging in size from 3 to 1,105 kilobases (kb), SpeI yielded 16 fragments (12 to 1,645 kb), DraI yielded at least 25 fragments (6 to 800 kb), and SnaBI generated 10 fragments (12 to 1,225 kb). A total genome size of approximately 4,400 +/- 112 kb was determined by summing the fragment lengths in each of the digests generated by using the different restriction endonucleases. The total genomic DNA consisted of chromosomal DNA (3,960 +/- 112 kb) and the five endogenous plasmids (approximately 450 kb total) whose cognate DNA fragments have been unambiguously identified. A number of genes have been physically mapped to the AseI-generated restriction endonuclease fragments of total genomic DNA by Southern hybridization analysis with either homologous or heterologous specific gene probes or, in the case of several auxotrophic and pigment-biosynthetic mutants apparently generated by Tn5, a Tn5-specific probe. Other genes have been mapped by a comparison with wild-type patterns of the electrophoretic banding patterns of the AseI-digested genomic DNA derived from mutants generated by the insertion of either kanamycin or spectinomycin-streptomycin resistance cartridges. The relative orientations, distance, and location of the pufBALMX, puhA, cycA, and pucBA operons have also been determined, as have been the relative orientations between prkB and hemT and between prkA and the fbc operon. PMID- 2553663 TI - Fine-structure genetic map of the maltose transport operon of Salmonella typhimurium. AB - We have constructed a fine-structure genetic map of the maltose transport operon in Salmonella typhimurium. We have isolated mal mutants by using indicator plates, penicillin selection, or a proton suicide technique. Mutants were obtained as spontaneous events or were induced by chemical mutagenesis and transposon insertion. Tn10 and Mu d(lac Ap)1 insertion mutations were used to create deletions. Mutations were also obtained in a gene that is equivalent to lamB in Escherichia coli, which codes for the lambda bacteriophage receptor. The gene products in the mutants were characterized by sodium dodecyl sulfate polyacrylamide-gel electrophoresis and immunoblotting. Our data indicate that the location of this operon on the Salmonella chromosome as well as the gene order and its orientation are the same as those in E. coli. This map will be useful in studying the mechanism of periplasmic transport in S. typhimurium. PMID- 2553664 TI - Export of FepA::PhoA fusion proteins to the outer membrane of Escherichia coli K 12. AB - A library of fepA::phoA gene fusions was generated in order to study the structure and secretion of the Escherichia coli K-12 ferric enterobactin receptor, FepA. All of the fusion proteins contained various lengths of the amino terminal portion of FepA fused in frame to the catalytic portion of bacterial alkaline phosphatase. Localization of FepA::PhoA fusion proteins in the cell envelope was dependent on the number of residues of mature FepA present at the amino terminus. Hybrids containing up to one-third of the amino-terminal portion of FepA fractionated with their periplasm, while those containing longer sequences of mature FepA were exported to the outer membrane. Outer membrane localized fusion proteins expressed FepA sequences on the external face of the outer membrane and alkaline phosphatase moieties in the periplasmic space. From sequence determinations of the fepA::phoA fusion joints, residues within FepA which may be exposed on the periplasmic side of the outer membrane were identified. PMID- 2553665 TI - Characterization of an insertion sequence (IS891) of novel structure from the cyanobacterium Anabaena sp. strain M-131. AB - When recombinant plasmids that were transferred to the cyanobacterium Anabaena sp. strain M-131 were transferred back to Escherichia coli, some of the transformants contained inserts. One of the insertion sequences (ISs) was characterized by sequencing. This 1,351-base-pair IS contained an open reading frame that was capable of encoding a peptide of 310 amino acids and had terminal sequences with distinctive structures, but it lacked terminal inverted repeats and did not duplicate target DNA upon insertion. The element bore no significant sequence homology to any sequence stored in the GenBank data base. Restriction analysis of the genomes of Anabaena sp. strain M-131 and Anabaena sp. strain PCC 7120 showed those strains to be closely related. Sequences homologous to the IS element were also present in the DNA of Anabaena strain PCC 7120, but the copy numbers and chromosomal locations of such sequences differed in the two strains. The largest visualized plasmid was 425 kilobases (kb) in M-131 and 410 kb in PCC 7120; at least the former plasmid contained multiple copies of the element, as did a 115-kb plasmid in M-131. PMID- 2553666 TI - Differential activity of a transposable element in Escherichia coli colonies. AB - In Escherichia coli colonies, patterns of differential gene expression can be visualized by the use of Mu d(lac) fusion elements. Here we report that patterned beta-galactosidase expression in colonies of strain MS1534 resulted from a novel mechanism, spatially localized replication of the Mu dII1681 element causing lacZ transposition to active expression sites. Mu dII1681 replication did not occur constitutively with a fixed probability but was dependent on the growth history of the bacterial population. The bacteria in which Mu dII1681 replication and lacZ transposition had occurred could no longer form colonies. These results lead to several interesting conclusions about cellular differentiation during colony development and the influence of bacterial growth history on gene expression and genetic change. PMID- 2553667 TI - Genetic analysis of the conjugal transfer determinants encoded by the streptococcal broad-host-range plasmid pIP501. AB - pIP501 is a 30.2-kilobase (kb) broad-host-range conjugative streptococcal plasmid which encodes chloramphenicol and erythromycin resistance. A smaller conjugation proficient derivative of pIP501 has been constructed and designated pVA1702. pVA1702 is 25.2 kb, encodes only kanamycin resistance, and confers conjugative ability in an Enterococcus faecalis mating system. The temperature-sensitive transposon delivery vectors pTV1ts and pTV32 carrying Tn917 and Tn917lac, respectively, were used to create insertion mutants of pVA1702 in E. faecalis. These insertions enabled us to localize the determinants conferring conjugation to two separate regions of pVA1702: the 7.5-kb region A and the 8.8-kb region B. A nested set of deletions of the 7.5-kb region was subcloned in Escherichia coli, and the expression of these deletions was examined in an E. coli minicell system. The genes for three conjugal proteins have been located and designated cnjA, cnjB, and cnjC (for conjugation). Two other peptides that are transcribed in a divergent direction from the cnj genes were encoded in the A region, but it is not known whether they are involved in conjugation. These studies indicate that the conjugation genes of pIP501 are arranged in multiple transcriptional units. PMID- 2553668 TI - Purification and some properties of glutathione S-transferase from Escherichia coli B. AB - Glutathione S-transferase was purified approximately 2,300-fold from cell extracts of Escherichia coli B with a 7.5% activity yield. The molecular weight of the enzyme was 45,000, and the enzyme appeared to consist of two homogeneous subunits. The enzyme was almost specific to 1-chloro-2,4-dinitrobenzene (Km, 1.43 mM) and glutathione (Km, 0.33 mM). The optimal pH and optimal temperature for activity were 7.0 and 50 degrees C, respectively, and the enzyme was stable from pH 5 to 11. The activity of the enzyme for 1-chloro-2,4-dinitrobenzene (3,2 mumol/min per mg of protein) was significantly lower than those of the enzymes from mammals, plants, and fungi. PMID- 2553669 TI - Molecular cloning and characterization of comC, a late competence gene of Bacillus subtilis. AB - comC is a Bacillus subtilis gene required for the development of genetic competence. We have cloned a fragment from the B. subtilis chromosome that carries comC and contains all the information required to complement a Tn917lac insertion in comC. Genetic tests further localized comC to a 2.0-kilobase HindIII fragment. Northern (RNA) blotting experiments revealed that an 800-base-pair comC specific transcript appeared at the time of transition from exponential to stationary phase during growth through the competence regimen. The DNA sequence of the comC region revealed two open reading frames (ORFs), transcribed in the same direction. The upstream ORF encoded a protein with apparent sequence similarity to the folC gene of Escherichia coli. Insertion of a chloramphenicol resistance determinant into this ORF and integration of the disrupted construct into the bacterial chromosome by replacement did not result in competence deficiency. The downstream ORF, which contained the Tn917lac insertion that resulted in a lack of competence, is therefore the comC gene. The predicted protein product of comC consisted of 248 amino acid residues and was quite hydrophobic. The comC gene product was not required for the expression of any other com genes tested, and this fact, together with the marked hydrophobicity of ComC, suggests that it may be a component of the DNA-processing apparatus of competent cells. PMID- 2553670 TI - The cytochrome bc1 complex of Rhodobacter sphaeroides can restore cytochrome c2 independent photosynthetic growth to a Rhodobacter capsulatus mutant lacking cytochrome bc1. AB - Plasmids encoding the structural genes for the Rhodobacter capsulatus and Rhodobacter sphaeroides cytochrome (cyt) bc1 complexes were introduced into strains of R. capsulatus lacking the cyt bc1 complex, with and without cyt c2. The R. capsulatus merodiploids contained higher than wild-type levels of cyt bc1 complex, as evidenced by immunological and spectroscopic analyses. On the other hand, the R. sphaeroides-R. capsulatus hybrid merodiploids produced only barely detectable amounts of R. sphaeroides cyt bc1 complex in R. capsulatus. Nonetheless, when they contained cyt c2, they were capable of photosynthetic growth, as judged by the sensitivity of this growth to specific inhibitors of the photochemical reaction center and the cyt bc1 complex, such as atrazine, myxothiazol, and stigmatellin. Interestingly, in the absence of cyt c2, although the R. sphaeroides cyt bc1 complex was able to support the photosynthetic growth of a cyt bc1-less mutant of R. capsulatus in rich medium, it was unable to do so when C4 dicarboxylic acids, such as malate and succinate, were used as the sole carbon source. Even this conditional ability of R. sphaeroides cyt bc1 complex to replace that of R. capsulatus for photosynthetic growth suggests that in the latter species the cyt c2-independent rereduction of the reaction center is not due to a structural property unique to the R. capsulatus cyt bc1 complex. Similarly, the inability of R. sphaeroides to exhibit a similar pathway is not due to some inherent property of its cyt bc1 complex. PMID- 2553671 TI - Constitutive activation of the fucAO operon and silencing of the divergently transcribed fucPIK operon by an IS5 element in Escherichia coli mutants selected for growth on L-1,2-propanediol. AB - L-1,2-Propanediol is an irretrievable end product of L-fucose fermentation by Escherichia coli. Selection for increased aerobic growth rate on propanediol results in the escalation of basal synthesis of the NAD+-linked oxidoreductase encoded by fucO, a member of the fuc regulon for the utilization of L-fucose. In general, when fucO becomes constitutively expressed, two other simultaneous changes occur: the fucA gene encoding fuculose-1-phosphate aldolase becomes constitutively expressed and the fucPIK operon encoding fucose permease, fucose isomerase, and fuculose kinase becomes noninducible. In the present study, we show that fucO and fucA form an operon which is divergently transcribed from the adjacent fucPIK operon. In propanediol-positive and fucose-negative mutants the cis-controlling region shared by the operons fucAO and fucPIK is lengthened by 1.2 kilobases. DNA hybridization identified the insertion element to be IS5. This element, always oriented in the same direction with the left end (the BglII end) proximal to fucA, apparently causes constitutive expression of fucAO and noninducibility of fucPIK. The DNA of the fucAO operon and a part of the adjacent fucP was sequenced. PMID- 2553672 TI - Genes involved in lipopolysaccharide production and symbiosis are clustered on the chromosome of Rhizobium leguminosarum biovar viciae VF39. AB - Four mutants of Rhizobium leguminosarum biovar viciae VF39 altered in lipopolysaccharide (LPS) synthesis were isolated upon random Tn5 mutagenesis. These mutants produced matt colonies on TY medium and showed autoagglutination and loss of motility. On sodium dodecyl sulfate-polyacrylamide gels, they lacked a slow-migrating carbohydrate band, corresponding to the complete LPS (LPSI). All four mutants formed small white nodules on Vicia hirsuta. These nodules were infected but showed no nitrogen-fixing activity and senesced prematurely. Three of the mutants were complemented by a wild-type cosmid to synthesis of normal LPS and induction of nitrogen-fixing nodules. By hybridization and in vivo cloning experiments, the mutations were mapped within different EcoRI fragments which could be localized on the VF39 chromosome. Cross-complementation analyses revealed that the three mutants were affected in different transcriptional units. The results indicate that a cluster of genes necessary for LPSI production and symbiotic efficiency is located within a defined region of 20 kilobases on the R. leguminosarum bv. viciae chromosome. PMID- 2553673 TI - Evidence that the Myxococcus xanthus frz genes are developmentally regulated. AB - The frizzy (frz) mutants of Myxococcus xanthus are unable to form fruiting bodies. Instead of forming discrete mounds, these strains aggregate as filaments which have a circular and tangled appearance. Mutations leading to this phenotype have been mapped to five complementation groups, frzA, frzB, frzCD, frzE, and frzF. All have been found to be involved in the control of directional movement of the bacteria and, except for frzB, to be homologous to the chemotaxis genes of enteric bacteria. In this report we present a study of the regulation of expression of the first four genes of the frz gene cluster (frzA, frzB, frzCD, and frzE) by using Tn5-lac transcriptional fusions as reporters of gene expression. We found that these frz genes are developmentally regulated, with their transcription peaking at about the time of early mound formation (12 to 18 h). Analysis of FrzCD expression by enzyme-linked immunosorbent assay showed a 10 fold greater induction at 15 h of development over the level of vegetative cell expression. Northern blot hybridization analysis suggested that the frz genes were arranged as an operon. To test this hypothesis, double mutants were constructed which contained Tn5-132 either upstream or downstream of the reporter Tn5-lac. The expression of the frz genes in the double mutants was consistent with the hypothesis that the first four genes (frzA, frzB, frzCD, and frzE) are organized as an operon with an internal promoter. Insertion mutations in frzCD lowered gene expression whether they were upstream or downstream of the reporter Tn5-lac, suggesting that the FrzCD protein regulates transcription of the entire operon from a promoter upstream of frzA. Evidence is presented suggesting that FrzE is required for induction of transcription as well. When frz mutations were placed in strains that were unable to aggregate (tag), the frz genes were expressed at an elevated level on fruiting agar; this high level of expression was maintained for several days. These results suggest that the tag gene products interact with the frz functions. PMID- 2553674 TI - Characterization of the cryptic lambdoid prophage DLP12 of Escherichia coli and overlap of the DLP12 integrase gene with the tRNA gene argU. AB - The argU (dnaY) gene of Escherichia coli is located, in clockwise orientation, at 577.5 kilobases (kb) on the chromosome physical map. There was a cryptic prophage spanning the 2 kb immediately downstream of argU that consisted of sequences similar to the phage P22 int gene, a portion of the P22 xis gene, and portions of the exo, P, and ren genes of bacteriophage lambda. This cryptic prophage was designated DLP12, for defective lambdoid prophage at 12 min. Immediately clockwise of DLP12 was the IS3 alpha 4 beta 4 insertion element. The argU and DLP12 int genes overlapped at their 3' ends, and argU contained sequence homologous to a portion of the phage P22 attP site. Additional homologies to lambdoid phages were found in the 25 kb clockwise of argU. These included the cryptic prophage qsr' (P. J. Highton, Y. Chang, W. R. Marcotte, Jr., and C. A. Schnaitman, J. Bacteriol. 162:256-262, 1985), a sequence homologous to a portion of lambda orf-194, and an attR homolog. Inasmuch as the DLP12 att int xis exo P/ren region, the qsr' region, and homologs of orf-194 and attR were arranged in the same order and orientation as the lambdoid prophage counterparts, we propose that the designation DLP12 be applied to all these sequences. This organization of the DLP12 sequences and the presence of the argU/DLP12 int pair in several E. coli strains and closely related species suggest that DLP12 might be an ancestral lambdoid prophage. Moreover, the presence of similar sequences at the junctions of DLP12 segments and their phage counterparts suggests that a common mechanism could have transferred these DLP12 segments to more recent phages. PMID- 2553675 TI - Isolation and characterization of a laccase-derepressed mutant of Neurospora crassa. AB - Laccase from the ascomycete Neurospora crassa is an inducible secretory enzyme. Production of this enzyme is repressed in vegetative cultures but can be induced by treatment with low concentrations of cycloheximide. Isolation and characterization of a derepressed mutant, the lah-1 mutant, that is capable of producing laccase in vegetative cultures without induction by cycloheximide are described. The lah-1 mutation is mapped between nit-2 and leu-3 on linkage group I, and it behaved as a recessive mutation in a forced heterokaryon. No differences were detected biochemically or immunologically between the laccase protein produced by the lah-1 mutant in the absence of cycloheximide and that induced with cycloheximide in the wild-type strain. This suggests that both laccases (66 kilodaltons) are products of the same structural gene. Relative amounts of laccase in the culture filtrate of the lah-1 mutant were much higher than those induced with cycloheximide in the wild-type strain, demonstrating high efficiency of the lah-1 mutant in production and secretion of laccase. The time course of laccase production by the lah-1 mutant revealed that expression of 66 kilodalton laccase was repressed in conidia and derepressed during vegetative mycelial growth. This suggests that a multiple regulatory mechanism is involved in the production and/or maturation of Neurospora laccase. The lah-1 mutant may be useful for identifying genes that regulate expression of the laccase gene in N. crassa. PMID- 2553676 TI - Mutational analysis of the histidine operon promoter of Salmonella typhimurium. AB - We isolated a collection of 67 independent, spontaneous Salmonella typhimurium his operon promoter mutants with decreased his expression. The mutants were isolated by selecting for resistance to the toxic lactose analog o-nitrophenyl beta-D-thiogalactoside in a his-lac fusion strain. The collection included base pair substitutions. small insertions, a deletion, and one large insertion identified as IS30 (IS121), which is resident on the Mu d1 cts(Apr lac) phage used to construct the his-lac fusion. Of the 37 mutations that were sequenced, 14 were unique. Six of the 14 were isolated more than once, with the IS30 insertion occurring 16 times. The mutations were located throughout the his promoter region, with two in the conserved - 35 hexamer sequence, four in the conserved - 10 hexamer sequence (Pribnow box), seven in the spacer between the - 10 and -35 hexamer sequences, and the IS30 insertions just upstream of the -35 hexamer sequence. Four of the five substitution mutations changed a consensus base pair recognized by E sigma 70 RNA polymerase in the -10 or -35 hexamer. Decreased his expression caused by the 14 different his promoter mutations was measured in vivo. Relative to the wild-type promoter, the mutations resulted in as little as a 4-fold decrease to as much as a 357-fold decrease in his expression, with the largest decreases resulting from changes in the most highly conserved features of E sigma 70 promoters. PMID- 2553677 TI - The bvgA gene of Bordetella pertussis encodes a transcriptional activator required for coordinate regulation of several virulence genes. AB - The bvg region of the respiratory pathogen Bordetella pertussis coordinately regulates the expression of several unlinked virulence determinants in response to environmental signals. The DNA sequence of the bvg region contains three genes (bvgA, bvgB, and bvgC). Transcription of a single-copy fusion consisting of the upstream region of a bvg-activated B. pertussis gene (fhaB) attached to the promoterless lac operon in Escherichia coli requires the entire bvgABC region in trans. Activation of the fhaB::lacZYA fusion is sensitive to the same environmental stimuli in E. coli that modulate the expression of bvg-activated genes in B. pertussis. Our data show that overexpression of the bvgA gene from a strong heterologous promoter results in transcriptional activation of the fhaB::lacZYA fusion even in the absence of the bvgB and bvgC products. Activation of fhaB transcription by bvgA overexpression in E. coli is no longer repressed by environmental conditions. The bvgA product has been identified by maxicell analysis as a 23-kilodalton protein. A B. pertussis mutant containing an in-frame deletion in bvgA was constructed. This mutant was nonhemolytic and no longer produced filamentous hemagglutinin and pertussis toxin. The mutation in this strain was complemented by returning the bvgA gene in trans. Transcriptional chloramphenicol acetyltransferase fusions to the fhaB and ptx promoter regions were returned to both the B. pertussis bvgA deletion mutant and its parental wild type strain. Analysis of these strains indicated that the deletion mutant was defective in transcription of both ptx and fhaB. We conclude from these data that bvgA, bvgB, and bvgC comprise an operon encoding the components essential for coordinate regulation and sensory transduction. The BvgA protein is a transcriptional regulatory factor. The bvgB and bvgC products may be important in regulating the activity of BvgA in response to the changing environmental stimuli that B. pertussis encounters during the diseases whooping cough. PMID- 2553678 TI - Analysis of Bordetella pertussis virulence gene regulation by use of transcriptional fusions in Escherichia coli. AB - The virulence regulon of Bordetella pertussis includes a trans-acting regulatory locus, bvg, that is required for expression of several virulence factors. The virulence control system also responds to environmental signals. We have reconstructed a bvg-dependent regulatory system in Escherichia coli by using bacteriophage lambda vectors carrying transcriptional fusions to lacZYA. Single copy lacZYA fusions to the B. pertussis fhaB locus, which encodes the attachment factor filamentous hemagglutinin, were activated nearly 400-fold by pBR322 replicons carrying sequences that included bvg. In contrast, bvg had no effect on the pertussis toxin operon (ptxA-E) promoter in E. coli as measured by ptxA-lacZ expression. Environmental signals that modulate expression of virulence genes in B. pertussis had a pronounced effect on bvg-mediated activation of fhaB-lacZ. MgSO4, nicotinic acid, and low temperature resulted in decreases in beta galactosidase activities of 175-, 115-, and 45-fold respectively. Sensory transduction and transcriptional activation were tightly coupled, and both required an intact bvg locus as determined by 5' and 3' deletions that eliminated both activities. PMID- 2553679 TI - Inhibition of growth of ftsQ, ftsA, and ftsZ mutant cells of Escherichia coli by amplification of a chromosomal region encompassing closely aligned cell division and cell growth genes. AB - Amplification of a 2.6-kilobase chromosomal fragment of the mra region of Escherichia coli encompassing the ftsI(pbpB) gene and an open reading frame upstream with lethal to E. coli strains with mutations of the flanking cell division genes ftsQ, ftsA, and ftsZ. A shortened fragment in which the major portion of ftsI was deleted also had lethal effects on ftsQ and ftsZ mutants. PMID- 2553680 TI - The structurally related exbB and tolQ genes are interchangeable in conferring tonB-dependent colicin, bacteriophage, and albomycin sensitivity. AB - Double exbB tolQ mutants of Escherichia coli were completely resistant to bacteriophages T1 and phi 80, in contrast to strains with exbB or tolQ mutations, which were sensitive. Cells carrying mutations in exbB were partially tolerant to colicins B, D, and M and became fully tolerant by the introduction of tolQ mutations. This suggested involvement of both exbB and tolQ in tonB-dependent uptake. PMID- 2553681 TI - Stimulatory effects of a short chain phosphatidate on superoxide anion production in guinea pig polymorphonuclear leukocytes. AB - Treatment of guinea pig polymorphonuclear leukocytes (PMNL) with a phosphatidate containing short-chain fatty acids, 1,2-didecanoyl-3-sn-phosphatidate (PA10), induced substantial superoxide anion (O2-) production in a dose-dependent manner, whereas phosphatidates prepared from egg lecithin and 1,2-dioleoyl-3-sn phosphatidate (PA18:1) had no such effect. Calcium was not involved in PA10 induced O2- production, since the production was also observed in the case of addition of EGTA prior to PA10 or pretreatment of PMNL with quin-2 and EGTA to eliminate contributions of both extracellular and intracellular calcium. We have reported in previous papers that the phosphorylation of 46K protein(s), which was commonly observed in parallel with an activation of NADPH oxidase in PMNL, was increased by treatment with 10 microM 1-oleoyl-2-acetylglycerol (OAG) with little change in the O2- production (Okamura et al. (1984) Arch. Biochem. Biophys. 228, 270-277; Ohtsuka et al. (1988) Arch. Biochem. Biophys. 260, 226-231). Treatment of PMNL with a combination of PA10, which slightly increased 46K protein phosphorylation, and such a low concentration of OAG induced a marked increase in the O2- production with the increase in 46K protein phosphorylation, which was probably due to OAG action. Thus, it is likely that this protein phosphorylation plays a significant role in the stimulation of the O2- production by phosphatidate in PMNL. PMID- 2553682 TI - Identification and characterization of inhibitory sequences in four repeating domains of the endogenous inhibitor for calcium-dependent protease. AB - We reported previously the cDNA cloning of the endogenous inhibitor for calcium dependent protease (CANP inhibitor, calpastatin) and the expression of its fragments in Escherichia coli. The CANP inhibitor has four internal repeating domains each spanning about 140 amino acid residues. The inhibitory activity arises from these domains which have a well-conserved sequence, TIPPXYR, in their central positions. The inhibitory activities of various fragments expressed in E. coli suggest the involvement of the regions around the well-conserved sequences. In this report, we describe further detailed investigation on the interaction site of the CANP inhibitor with CANP by truncating inhibitor fragments and by using chemically synthesized peptides. The results clearly indicate that the sequence around the well-conserved sequence, TIPPXYR, is an interaction site. A peptide as short as 23 amino acid residues retained inhibitory activity, but a 9 residue peptide corresponding to the conserved sequence, VTIPPKYRE had none. The inhibitory sequence is suggested as LGXKDREXTIPPXYRXLL. The analysis of the competition between an inhibitor peptide and an irreversible inhibitor, E-64 for the reaction with the active site suggests no involvement of the active site cysteine residue of CANP in the inhibitory interaction between CANP and the CANP inhibitor. The high specificity of the CANP inhibitor to CANP arises from its interaction with residues other than the active site cysteine residue, possibly the subsite for substrate-binding of CANP. PMID- 2553683 TI - A fragmin-like protein from plasmodium of Physarum polycephalum that severs F actin and caps the barbed end of F-actin in a Ca2+-sensitive way. AB - Many protein factors regulating actin polymerization can be extracted from plasmodia of Physarum polycephalum in the presence of a high EGTA concentration (30 mM). A protein factor with the molecular weight of 60,000 (60 kDa protein) was especially interesting because of its fragmin-like properties. We purified and characterized this 60 kDa protein in the present study. The purified 60 kDa protein enhanced the initial rate of G-actin polymerization, severed F-actin, and capped the barbed end of F-actin in a Ca2+-dependent way. The threshold concentration for Ca2+ was around 10(-6) M. The flow birefringence measurement showed that the length of F-actin decreased from 2.8 to 1.0 microns depending on the concentration of 60 kDa protein added to F-actin. These properties were identical to those of fragmin (Mr 42,000) isolated from plasmodia (Hasegawa et al. (1980) Biochemistry 19, 2677-2683). However, the molecular weight, the tryptic peptide map, and the cross-reactivities with polyclonal anti-fragmin antibodies were different from those of fragmin. We concluded from these results that 60 kDa protein is a new Ca2+-sensitive F-actin-severing protein. Considering its similarity to fragmin, we termed the 60 kDa protein fragmin 60. PMID- 2553684 TI - Rubredoxin from Clostridium perfringens: complete amino acid sequence and participation in nitrate reduction. AB - The complete primary structure of rubredoxin (Rd) isolated from Clostridium perfringens was sequenced to be: MKKFICDVCGYIYDPAVGDPDNGVEPGTEFKDIPDDWVCPLCGVDKSQFSETEE. The sequence was highly homologous to that of C. pasteurianum Rd but was different at 13 sites out of the total 54 amino acid residues (76% homology). It contained 1 Fe atom, 4 cysteine residues, and no labile sulfur, had a molecular weight of 6,056, and shared the general properties of classical anaerobic Rds. The pI was 4.4. The Rd was reduced with NADH in the presence of a specific NAD(P)H oxidoreductase preparation from the bacterium. The Km value of nitrate reductase for Rd as an electron-donor was 12 microM, a value comparable to that of the 13 microM for ferredoxin (Fd). These results taken together provide additional support for its role as the electron carrier in the nitrate reductase system [Seki, S., Ikeda, A., and Ishimoto, M. (1988) J. Biochem. 103, 583-584]. PMID- 2553686 TI - Simulation of natural enamel caries in vitro with methylcellulose acid gels: effect of addition of calcium and phosphate ions. AB - A modified acid gel technique for producing caries-like lesions in dental enamel has been developed. The decalcifying medium consists of a 10% (w/v) methylcellulose gel acidified with 0.1 M lactic acid-sodium lactate at pH 4.5 and having a hydroxyapatite content of 0.05% (w/v). This medium can produce caries like lesions in vitro at a slow rate, which are indistinguishable from natural enamel caries when examined by polarised light microscopy and contact microradiography. PMID- 2553685 TI - Chloroplast thylakoid proteins associated with sequestered proton-buffering domains. Plastocyanin contributes buffering groups to localized proton domains. AB - Thylakoid membrane proteins are organized so as to shield 30-50 nmol H+ (mg Chl) 1 from freely equilibrating with either the external or the lumen aqueous phases. Amine groups provide binding sites for this metastable buffering array and can be quantitatively measured by acetylation using [3H]acetic anhydride. The principle of the assay is that a metastable acidic domain will have relatively less of the reactive neutral form of the amine compared to the amount present after addition of an uncoupler. The extent of the acetylation reaction is strongly influenced by whether the lumen pH comes to complete equilibrium with the external pH prior to adding the acetic anhydride. Determination of the lumen pH by [14C]methylamine distribution after the standard 3 or 5 min equilibration in pH 8.6 buffer indicated that the lumen may have been 0.2 to 0.3 pH more acidic than the external phase. This effect was taken into account by determining the pH dependence, in the pH 8.2-8.6 range, of acetylation of the membrane proteins studied, and the labeling data were conservatively corrected for this possible contribution. Experiments were carried out to identify the thylakoid proteins that contribute such metastable domain amine groups, using the above conservative correction. Surprisingly, plastocyanin contributes buried amine groups, but cytochrome f did not give evidence for such a contribution, if the conservative correction in the labeling was applied. If the correction was too conservative, cytochrome f may contribute amines to the sequestered domains. The new methodology verified earlier results suggesting that three Tris-releasable photosystem II-associated proteins also contribute significantly to the sequestered amine-buffering array. PMID- 2553687 TI - The hydroxyapatite-binding regions of a rat salivary glycoprotein. AB - The regions of a salivary sulphated glycoprotein which are involved in its attachment to hydroxyapatite (Biogel HTP) have been characterised. The sulphated glycoprotein, a 35S-labelled preparation from mixed palatal and buccal minor gland secretions of the rat was bound onto hydroxyapatite and the resultant glycoprotein-hydroxyapatite complex was sequentially digested with pronase E and alpha-L-fucosidase, a treatment which released 86.8% +/- 1.7% of the radioactivity of the initially bound glycoprotein. The fragments which remained attached to the hydroxyapatite after enzymic digestion were fractionated on Sephadex G-25 and analysed for carbohydrate and amino acid components. A range of amino acids were detected which could reflect both glycosylated and non glycosylated-binding regions. Sialic acid, although considered to be involved in the attachment process was not detected in any of the fragments remaining after enzymic digestion, a finding which provides indirect evidence that the enzymically liberated products do not subsequently re-attach to the hydroxyapatite surface. The notable feature of the fractions with average Mr estimated at 1000 or less is the high proportion of N-acetylhexosamine and N acetylgalactosamine. It is apparent that the hexosamine residues, which normally bear the ester sulphate moieties of sulphated glycoproteins, play an important role in the attachment of sulphated glycoproteins to hydroxyapatite. PMID- 2553688 TI - Isolation and characterization of three non-mucinous human salivary proteins with affinity for hydroxyapatite. AB - The isolation and chemical analysis of three human salivary proteins is reported. Using cation-exchange, FPLC anion-exchange chromatography and gel filtration, three human salivary proteins with affinity for hydroxyapatite were isolated, having molecular weights of 60 kD, 20 kD and 14 kD, respectively. The 60 kD protein was identified as albumin, and the 14 kD protein as a member of the cystatin family. Isoelectric focusing of the 14 kD protein revealed a single band, having an isoelectric point (pl) of 4.7. The third protein, not described yet, is appearing as a 20 kD doublet on SDS-PAGE. Isoelectric focusing resolved the 20 kD protein into four bands having pl's of 4.8, 5.0, 5.2 and 5.4, respectively. All bands were recognized by monoclonal antibodies to the 20 kD protein indicating that these four protein bands share the same epitope. The 20 kD protein is a glycoprotein with a carbohydrate content of 13% and a molar ratio of Fuc: Man: Gal: GlcNAc: NeuAc = 3.4:2.6:2.9:4.0:0.4. All three proteins bind strongly to a hydroxyapatite-based HPHT column at pH 6.0. With increasing pH, the binding diminished, especially of the 14 kD protein. PMID- 2553689 TI - Receptor-mediated transport of heme by hemopexin regulates gene expression in mammalian cells. AB - Hemopexin (HPX) transports heme to liver parenchymal cells, undergoes receptor mediated endocytosis, and recycles intact. Incubation of mouse hepatoma (Hepa) cells with heme-HPX causes a rapid dose- and time-dependent increase in the steady-state level of heme oxygenase (HO) mRNA. A maximum induction of 20-25-fold is achieved within 3 h after incubation with 10 microM heme-HPX. This accumulation of HO mRNA results primarily from increased transcription of the HO gene as judged by in vitro nuclear run-on assays. In addition, receptor-mediated transport of heme into Hepa cells significantly decreases the steady-state level of transferrin receptor (TfR) mRNA. While a 25-30-fold decrease in the amount of TfR mRNA is observed within 3 h of incubation of Hepa cells with 10 microM heme HPX, no significant change in the rate of TfR gene transcription was detected. These regulatory effects of heme-HPX are not restricted to hepatic cells but are also observed in human promyelocytic HL-60 cells. This is the first direct demonstration of receptor-mediated transport of heme by hemopexin regulating gene expression in mammalian cells. PMID- 2553690 TI - The cyclic AMP-dependent protein kinase from bovine cardiac muscle is a homoserine kinase. AB - The catalytic subunit of the cAMP-dependent protein kinase from bovine cardiac muscle phosphorylates homoserine in the synthetic peptide Leu-Arg-Arg-Ala-Hse-Leu Gly. Phosphorylation of the primary alcohol of the homoserine residue was established via NMR spectroscopy. Two-dimensional correlated and nuclear Overhauser effect spectroscopies provided the sequence-specific chemical shift assignments of the substrate peptide and its phosphorylated counterpart. Coupled and decoupled 31P NMR experiments established the presence of phosphate on the homoserine residue. The maximal velocity (6.4 mumol/min.mg) obtained for homoserine-peptide phosphorylation at 12.5 mM Mg2+ compares favorably to the velocities observed for the corresponding serine- (21 mumol/min.mg), threonine- (3.2 mumol/min.mg), and hydroxyproline-peptides (1 mumol/min.mg). However, the Km for homoserine kinase activity is modest (1.3 mM) relative to the Km associated with the phosphorylation of the serine-containing substrate (22 microM). The effect of Mg2+ concentration on the kinetic parameters kcat, Km, and kcat/Km was investigated for both serine- and homoserine-peptides. Both substrates display similar kcat/Km versus [Mg2+] profiles, with the most notable difference that the optimal Mg2+ concentration is higher for the homoserine-containing peptide. In addition, the Km for the serine-peptide was found to be independent of [Mg2+], whereas the Km for the homoserine-peptide was observed to be dependent upon [Mg2+]. These results suggest that the long homoserine side chain may induce an unusually large off rate for the peptide and/or may misalign the hydroxyl moiety in the active site. PMID- 2553691 TI - The potential role of DNA methylation in the response to 2,3,7,8 tetrachlorodibenzo-p-dioxin. AB - The Ah receptor is an intracellular protein that binds the environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin. The liganded receptor interacts with a specific DNA recognition motif located within a dioxin-responsive enhancer upstream of the CYP1A1 gene. Methylation protection and methylation interference studies indicate that the liganded receptor contacts both DNA strands, at 4 guanine residues contained within the recognition motif. These findings imply that the liganded receptor interacts with its cognate enhancer within the major groove of the DNA helix. Cytosine methylation of the recognition motif at CpG dinucleotides diminishes the protein-DNA interaction, as measured by gel retardation. Furthermore, methylation at cytosine inhibits the enhancer function of the DNA. These findings imply that DNA methylation can diminish the response to dioxin by impeding the Ah receptor-enhancer interaction. PMID- 2553692 TI - Generation of Lys-gamma 3-melanotropin from pro-opiomelanocortin 1-77 by a bovine intermediate lobe secretory vesicle membrane-associated aspartic protease and purified pro-opiomelanocortin converting enzyme. AB - The ability of bovine intermediate lobe secretory vesicle membrane-associated enzyme(s) and purified, soluble paired basic residue-specific, pro opiomelanocortin converting enzyme (Loh, Y.P., Parish, D. C., and Tuteja, R. (1985) J. Biol. Chem. 260, 7194-7205) to cleave bovine NH2-terminal pro opiomelanocortin1-77 (N-POMC 1-77) was investigated. Purified pro opiomelanocortin converting enzyme and an enzyme activity associated with the secretory vesicle membrane were shown to cleave bovine N-POMC1-77 to two major products: N-POMC1-49 and Lys-gamma 3-melanotropin (MSH), and one minor product, gamma 3-MSH. These products were identified by their retention times on high performance liquid chromatography, immunological characteristics, and for Lys gamma 3-MSH, amino acid composition. The products generated indicate cleavage preferentially between Arg 49-Lys 50 of bN-POMC1-77 (where b indicates bovine), which is identical to the processing pattern found in the bovine intermediate lobe in situ. The membrane converting activity was shown to be stimulated by 5 mM Ca2+ and has a pH optimum of 4-5 and an inhibitor profile characteristic of an aspartic protease. This suggests that the membrane-associated enzyme involved is very similar or identical to the purified, soluble pro-opiomelanocortin converting enzyme, which has previously been reported to be an acidic, aspartic protease responsible for the initial steps of POMC processing. The results of this study lead to the proposal that the lack of processing of the Arg49-Lys50 site in POMC in the anterior lobe versus the intermediate lobe of the pituitary in vivo may be due to other regulatory mechanisms rather than invoking the existence in the intermediate lobe of another enzyme specific for this site, different from pro-opiomelanocortin converting enzyme. PMID- 2553693 TI - Polyphosphoinositides produced by phosphatidylinositol 3-kinase are poor substrates for phospholipases C from rat liver and bovine brain. AB - The ability of three pure types of bovine brain phospholipase C (PLC) and one pure rat liver PLC to utilize as substrates the recently discovered phosphatidylinositol 3-phosphate (PI-3-P), a putative phosphatidylinositol 3,4 bisphosphate (PI-3,4-P2), and phosphatidylinositol trisphosphate (PIP3) was investigated. PI-3-P, PI-3,4-P2, and PIP3 are the products of phosphorylation of PI, PI-4-P, and PI-4,5-P2, respectively, by phosphoinositide 3-kinase activities that are associated with certain protein-tyrosine kinases. Although these new phospholipids have been found in intact cells, PI-3,4-P2 and PIP3 appear only after stimulation of quiescent cells with growth factors such as platelet-derived growth factor (Auger, K. R., Serunian, L. A., Soltoff, S. P., Libby, P., and Cantley, L. C. (1989) Cell 57, 167-175) and after transformation by certain oncoproteins (L. A. Serunian, K. R. Auger, T. M. Roberts, and L. C. Cantley, manuscript in preparation). Mixtures of [3H]PI-4-P plus [32P]PI-3-P or [3H]PI-4,5 P2 plus [32P]PI-3,4-P2 or PIP3 alone were used as substrates for PLCs in vitro. After incubation with enzyme followed by extraction with chloroform/methanol/HCl, the ratio of 3H/32P in the aqueous layer revealed the selective hydrolysis of PI 4-P and PI-4,5-P2 over PI-3-P and PI-3,4-P2. High performance liquid chromatography analysis of the aqueous layer containing reaction products confirmed that only PI-4-P and PI-4,5-P2, were hydrolyzed to inositol 1,4-P2 and inositol 1,4,5-P3, respectively. These findings suggest that the turnover of PI-3 P, PI-3,4-P2, and PIP3 occurs independently of the turnover of PI-4-P and PI-4,5 P2. PMID- 2553694 TI - Sodium ions as substitutes for protons in the gastric H,K-ATPase. AB - In view of the striking homology among various ion-translocating ATPases including Na,K-ATPase, Ca-ATPase, and H,K-ATPase, and the recent evidence that protons can replace cytoplasmic sodium as well as potassium in the reaction mechanism of the Na,K-ATPase (Polvani, C., and Blostein, R. (1988) J. Biol. Chem. 263, 16757-16763), we studied the role of sodium as a substitute for protons in the H,K-ATPase reaction. Using hog gastric H,K-ATPase-rich inside-out membrane vesicles we observed 22Na+ influx which was stimulated by intravesicular potassium ions (K+i) at pH 8.5 but not at pH 7.1. This sodium influx was observed in medium containing ATP and was inhibited by vanadate and SCH28080, a selective inhibitor of the gastric H,K-ATPase. At least 2-fold accumulation of sodium was observed at pH 8.5. Experiments aimed to determine the sidedness of the alkaline pH requirement for K+i-dependent sodium influx showed that K+i-activated sodium influx depends on pHout and is unaffected by changes in pHin. These results support the conclusion that sodium ions substitute for protons in the H,K-ATPase reaction mechanism and provide evidence for a similarity in ion selectivity and/or binding domains of the Na,K-ATPase and the gastric H,K-ATPase enzymes. PMID- 2553695 TI - A novel ATP-requiring protease from skeletal muscle that hydrolyzes non ubiquitinated proteins. AB - Previously, we isolated an ATP-dependent proteolytic pathway in muscle, liver, and reticulocytes that requires ubiquitin and the enzymes which conjugate ubiquitin to proteins. We report here that skeletal muscle contains another soluble alkaline energy-dependent (but ubiquitin-independent) proteolytic activity. The cleavage of non-ubiquitinated protein substrates by the partially purified protease requires ATP hydrolysis since ATP in the absence of Mg2+, nonhydrolyzable ATP analogs, and pyrophosphate all fail to stimulate proteolysis. Proteolytic activity is also stimulated by UTP, CTP, and GTP, although not as effectively as by ATP (Km(ATP) = 0.027 mM). The enzyme is inactivated by the serine protease inhibitors diisopropyl fluorophosphate and 3,4 dichloroisocoumarin, but not by specific inhibitors of aspartic, thiol, or metalloproteases. It is maximally active at pH 8 and has a molecular weight of approximately 600,000. This new activity differs from the 720-kDa multicatalytic proteinase, but resembles the soluble ATP-dependent proteolytic system that we previously isolated from murine erythroleukemia cells. PMID- 2553696 TI - Effect of Ca2+ on binding of the calpains to calpastatin. AB - Autolyzed mu-calpain, unautolyzed mu-calpain, autolyzed m-calpain, and unautolyzed m-calpain (mu-calpain is the micromolar Ca2+-requiring proteinase, m calpain is the millimolar Ca2+-requiring proteinase) were passed through a calpastatin-affinity column at different free Ca2+ concentrations, and binding of the calpains to calpastatin was compared with proteolytic activity of that calpain at each Ca2+ concentration. Unautolyzed m-calpain, autolyzed m-calpain, and autolyzed mu-calpain required less Ca2+ for half-maximal binding to calpastatin than for half-maximal activity. Unautolyzed mu-calpain, however, required slightly more Ca2+ for half-maximal binding to calpastatin than for half maximal activity. Half-maximal binding of oxidatively inactivated mu- or m calpain to calpastatin required approximately the same Ca2+ concentrations as half-maximal binding of unautolyzed mu- or m-calpain, respectively, to calpastatin. Binding of unautolyzed m-calpain and autolyzed mu-calpain to calpastatin occurred over a wide range of Ca2+ concentrations, and it seems likely that two or more Ca2+-binding sites with different Ca2+-binding constants are involved in binding of the calpains to calpastatin. Proteolytic activity occurs at different Ca2+ concentrations than calpastatin binding, suggesting a second set of Ca2+-binding sites associated with proteolytic activity. Third and fourth sets of Ca2+-binding sites may be involved in autolysis and in binding to phosphatidylinositol or cell membranes; these four Ca2+-dependent properties of the calpains may require the eight potential Ca2+-binding sites that amino acid sequences predict are present in the calpain molecules. PMID- 2553697 TI - Tissue-specific expression of four types of rat calmodulin-dependent protein kinase II mRNAs. AB - cDNA clones for a fifth polypeptide of rat brain calmodulin-dependent protein kinase II were isolated and sequenced. The cDNA sequence encoded a polypeptide, designated delta, consisting of 533 amino acid residues with a molecular weight of 60,080. Comparison of amino acid sequences of this and alpha, beta, beta', and gamma polypeptides of calmodulin-dependent protein kinase II reveals marked homology among them. The mRNAs for delta were expressed in rat brain tissues with different regional specificities. The distribution of alpha, beta/beta', gamma, and delta mRNAs in cerebrum, skeletal muscle, diaphragm, heart, small intestine, uterus, aorta, liver, kidney, lung, and testis were examined by RNA blot hybridization analysis with probes specific for the respective mRNAs. A 3.9 kilobase (kb) RNA species hybridizable with a probe for gamma was found in all the tissues examined, and 4.0-4.2-kb RNA species hybridizable with a probe for delta were found in all the tissues examined except for liver, while a 4.8-kb RNA species hybridizable with a probe for alpha and a 4.2-kb RNA species hybridizable with a probe for beta were present in brain but not in the other tissues. With the alpha probe, however, a 4.1- and 2.6-kb RNA species were both detected in skeletal muscle and diaphragm. With the beta probe, a 4.3-kb RNA in skeletal muscle and diaphragm, 2.9-kb RNA in small intestine, and 4.0-kb RNA in testis were detected. With the delta probe, a 3.5-kb RNA in heart and 1.8-kb RNA in testis were detected. Thus, gamma and delta mRNAs were expressed in various tissues, while alpha and beta/beta' mRNAs were primarily, if not exclusively, expressed in brain. PMID- 2553698 TI - Molecular cloning and DNA sequence analysis of Escherichia coli topB, the gene encoding topoisomerase III. AB - Escherichia coli contains two type 1 topoisomerases, topoisomerase I and III. Although topoisomerase III can be purified as a potent decatenase, its role in DNA metabolism is unclear. In order to address this issue, the gene encoding topoisomerase III from E. coli has been molecularly cloned and its DNA sequence determined. The cloned fragment of DNA contains an open reading frame that can encode a polypeptide of 73.2 kDa. The first 20 amino acids of this open reading frame are identical to those of topoisomerase III as determined by amino-terminal gas-phase microsequencing. Expression of the polypeptide encoded by this open reading frame, using a bacteriophage T7 transient expression system, results in the accumulation of a 74-kDa polypeptide. Soluble extracts prepared from cells overexpressing this gene product show a dramatic increase in topoisomerase activity when compared with control extracts. We propose that this gene be designated topB. PMID- 2553699 TI - Nuclear and nucleolar targeting sequences of c-erb-A, c-myb, N-myc, p53, HSP70, and HIV tat proteins. AB - Protein import into the cell nucleus requires specific binding of nuclear proteins to the nuclear pore complex. Based on amino acid sequence "motifs" of known nuclear targeting signals, we identified peptides within a number of nuclear proteins with likely nuclear targeting potential and tested their function by transfecting into cells fusion genes that produce the cytoplasmic "reporter" protein, pyruvate kinase (PK), joined to the test sequence. Sequences within c-myb (PLLKKIKQ), N-myc (PPQKKIKS), p53 (PQPKKKP), and c-erb-A (SKRVAKRKL) oncoproteins that direct PK hybrids into the nucleus were identified. A peptide (GRKKRRQRRRAP) of the human immunodeficiency virus (HIV) tat protein (Tat), which contains two short basic regions, targets fusion proteins to the nucleolus. The COOH-terminal basic Tat region (QRRRAP) does not target PK hybrid proteins into the nucleus, but mutation of two basic amino acids in this region decreases but does not abolish nucleolar accumulation mediated by the entire Tat nucleolar targeting sequence. Moreover, the c-Myc nuclear targeting sequence fused to the COOH-terminal basic Tat region (PAAKRVKLDQRRRAP) effectively localizes PK hybrids to the nucleus and nucleolus. A similar sequence (FKRKHKKDISQNKRAVRR) in the human heat-shock protein HSP70 also localizes PK to the nucleus and nucleolus. PMID- 2553700 TI - Inhibition of normal rat kidney cell growth by transforming growth factor-beta is mediated by collagen. AB - We have investigated the mechanism of inhibition of the serum-free monolayer growth of normal rat kidney (NRK) cells by transforming growth factor-beta (TGF beta). NRK cells grown on fibronectin-coated dishes exhibited a biphasic response to TGF-beta. Monolayer growth was slightly stimulated by subpicomolar concentrations, while picomolar concentrations of TGF-beta inhibited NRK cell growth in the presence or absence of epidermal growth factor. NRK cells exhibited a similar biphasic growth response to exogenous type I collagen. TGF-beta induced a 3-5-fold increase in the deposition of type I collagen-like proteins into the extracellular matrix of NRK cells during serum-free growth. Type I collagen-like proteins were identified by their sensitivity to degradation by purified bacterial collagenase and by Western blot analysis. The TGF-beta dose-response curves for induction of extracellular matrix-localized collagen and inhibition of NRK cell growth were similar. Finally, the inclusion of a purified bacterial collagenase, which did not degrade TGF-beta or TGF-beta receptors, or alter control NRK growth, prevented exogenous collagen or TGF-beta from inhibiting the serum-free growth of NRK cells. Our results demonstrate that an increase in collagen secretion plays an important role in the inhibition of the growth of NRK cells by TGF-beta. PMID- 2553701 TI - Interactions between tissue-type plasminogen activator and extracellular matrix associated plasminogen activator inhibitor type 1 in the human hepatoma cell line HepG2. AB - Hepatic parenchymal cells contribute to the clearance of circulating tissue-type plasminogen activator (t-PA) in vivo. The hepatocyte extracellular matrix is interposed between the endothelial-lined sinusoids and the parenchymal cell surface and thus may influence t-PA clearance. To test this hypothesis, the well differentiated human hepatoma cell line HepG2 was used to characterize the role of extracellular matrix in t-PA clearance in vitro. Previous studies with these cells demonstrated their capacity for specific catabolism of t-PA in a system modulated by plasminogen activator inhibitor type 1 (PAI-1). In the present study the extracellular matrix growth substratum of HepG2 cells is shown to contain active PAI-1. PAI-1 is distributed in a punctuate pattern throughout the substratum. Components of the substratum confer stability to active PAI-1 for intervals of at least 24 h. Exposing substratum to 125I-t-PA leads rapidly to the formation and release of a sodium dodecyl sulfate-stable 95-kDa 125I-t-PA.PAI-1 complex. In comparison, cell monolayers have the additional capacity for specific binding of the complex. However, PAI-1 is not detected at the surface of HepG2 cells in suspension, suggesting that 125I-t-PA.PAI-1 complexes form in substratum and subsequently bind to cells. Specific binding of performed 125I-t-PA.PAI-1, but not 125I-t-PA, was demonstrated for HepG2 cells in suspension. These results suggest that components of extracellular matrix participate in the clearance of t PA by hepatocytes. PMID- 2553702 TI - Beta 2-microglobulin enhances insulin-like growth factor I receptor levels and synthesis in bone cell cultures. AB - Beta 2-Microglobulin (beta 2m), a component of the major histocompatibility complex in chordates, has growth factor-like activity for cultured rat cells. We presently describe interactions between beta 2m and insulin-like growth factor I (IGF I) in osteoblast-enriched cultures. beta 2m increased DNA synthesis and was synergistic with IGF I. Affinity labeling revealed that beta 2m enhanced IGF I receptor number, and Northern analysis and radioimmunoassay showed that beta 2m increased steady state IGF I transcripts and medium IGF I polypeptide levels. These results indicate that the growth-promoting activity of beta 2m is mediated at least in part by regulating local IGF I binding and synthesis by skeletal cells. PMID- 2553703 TI - Carbachol in the presence of guanosine 5'-O-(3-thiotriphosphate) stimulates the breakdown of exogenous phosphatidylinositol 4,5-bisphosphate, phosphatidylinositol 4-phosphate, and phosphatidylinositol by rat brain membranes. AB - The breakdown of exogenously added [3H]inositol-labeled phosphoinositides by rat brain cortical membranes was stimulated by the muscarinic cholinergic agonist carbachol. The stimulation required the presence of guanine nucleotide. Optimal conditions were similar to those described for guanosine 5'-O-(3 thiotriphosphate) (GTP gamma S) + carbachol stimulation of phosphoinositide breakdown in [3H]inositol-prelabeled brain membranes (Claro, E., Garcia, A., and Picatoste, F. (1989) Biochem J. 261, 29-35). Carbachol stimulated [3H]phosphatidylinositol 4,5-bisphosphate (PIP2) breakdown was inhibited by atropine and guanosine 5'-O-(2-thiobisphosphate). The magnitude of the stimulation of exogenous PIP2 breakdown by carbachol and GTP gamma S (2- to 3 fold) was little affected over a PIP2 concentration range of 0.03-100 microM. Phosphatidylinositol 4-phosphate (PIP) was as good a substrate at all concentrations as PIP2 for carbachol stimulation of phospholipase C activity. There was appreciable phosphomonoesterase degradation of PIP to phosphatidylinositol (PI) over 10 min. There was also some conversion of added PIP to PIP2 in the presence of added ATP. The effect of calcium on PIP breakdown was similar to that on PIP2 breakdown, with an apparent EC50 for Ca2+ stimulation of 0.74 and 0.72 microM, respectively, under basal conditions. The stimulation of PIP2 and PIP breakdown by carbachol in the presence of GTP gamma S was greatest on a percentage basis at the lowest free Ca2+ concentrations. Above 1 microM free Ca2+, the stimulatory effect was lost, whereas 10 microM free Ca2+ gave a maximal stimulation of basal phospholipase C activity. Degradation of added PI was also stimulated by carbachol in the absence of ATP. PI breakdown had an EC50 for Ca2+ stimulation of 1.07 microM. The best stimulation of PI breakdown due to carbachol plus GTP gamma S was seen with 0.3 microM free Ca2+ and 100 microM PI. Maximal activation of PI breakdown was seen at 1 mM deoxycholate as was true for PIP2 and PIP breakdown. There was little effect, even of 30 microM GTP gamma S alone or of carbachol alone, on PI breakdown. Half-maximal activation of the carbachol response required only 0.2 microM GTP gamma S. These results indicate that the phospholipase C enzyme(s) activated by carbachol in the presence of GTP gamma S in rat brain cortical membranes can degrade PIP2, PIP, and PI to inositol phosphates and diacylglycerol. PMID- 2553704 TI - Investigation of the relationship between tyrosyl residues and the adenosine 5' monophosphate binding site of rabbit liver fructose-1,6-biphosphatase as studied by chemical modification and nuclear magnetic resonance spectroscopy. AB - The effects of AMP, fructose 6-phosphate (Fru-6-P), fructose 2,6-bisphosphate (Fru-2,6-P2), and paramagnetic ions on the aromatic region of the proton nuclear magnetic resonance (NMR) spectrum of rabbit liver fructose-1,6-bisphosphatase have been investigated at 300 MHz. Two well resolved peaks in this region of the NMR spectrum are assigned to the protons from the aromatic ring of a tyrosyl residue of the enzyme by chemical modification with tetranitromethane and by nuclear Overhauser effects. Nitration of the tyrosyl residue causes desensitization of the enzyme to AMP inhibition as well as the loss of activity. In the presence of AMP during the modifications, 1 tyrosyl residue could be protected, presumably the one observed by NMR. Binding of AMP, an allosteric inhibitor of the enzyme, to rabbit liver fructose-1,6-bisphosphatase leads to an upfield shift of the tyrosyl proton signals in the NMR spectrum. No chemical shift or line broadening could be detected in the presence of the paramagnetic manganous ion, Fru-2,6-P2, or Fru-6-P. The negative intramolecular nuclear Overhauser effect from the ribose H2' proton to the adenine H8 proton of AMP suggested that AMP binds to the enzyme with an anti conformation about the glycosidic bond. The failure to observe intermolecular nuclear Overhauser effects between the tyrosyl residue and the protons of AMP indicates that the distances between them are greater than 4 A. On the basis of these observations, it is suggested that the AMP-related tyrosyl residue may be close to the AMP binding site, but it is not directly involved in ligand binding. Rather, the protection of this tyrosyl residue by AMP as observed by chemical modification experiments may well be due to a conformational change that results from covalent modification of the enzyme. PMID- 2553705 TI - Multiple signaling pathways of histamine H2 receptors. Identification of an H2 receptor-dependent Ca2+ mobilization pathway in human HL-60 promyelocytic leukemia cells. AB - In order to analyze the complex activities of histamine H2 receptor activation on neutrophils, human HL-60 promyelocytic leukemia cells were differentiated into neutrophils by incubation with dimethyl sufoxide, loaded with the Ca2+-sensitive indicator dyes, indo-1 or fura-2, and the levels of intracellular Ca2+ ([Ca2+]i) measured in a fluorescent-activated cell sorter and fluorimeter, respectively. Histamine increased [Ca2+]i in a dose-dependent manner with a half-maximal concentration (EC50) of approximately 10(-6) to 10(-5) M, which exhibited H2 receptor specificity. Prostaglandin E2 and isoproterenol also induced [Ca2+]i mobilization in HL-60 cells, whereas the cell permeable form of cAMP and forskolin failed to increase [Ca2+]i. Since H2-receptor mediated [Ca2+]i mobilization was not inhibited by reducing the concentration of extracellular Ca2+ nor by the addition of Ca2+ channel antagonists, LaCl3 and nifedipine, [Ca2+]i mobilization is due to the release of Ca2+ from intracellular stores. Furthermore, both 10(-4) M histamine and 10(-6) M fMet-Leu-Phe increased the levels of 1,4,5-inositol trisphosphate. However, histamine-induced mobilization of [Ca2+]i was inhibited by cholera toxin but not by pertussis toxin, whereas the action of fMet-Leu-Phe was inhibited by pertussis toxin but not by cholera toxin. These data suggest that H2 receptors on HL-60 cells are coupled to two different cholera toxin-sensitive G-proteins and activate adenylate cyclase and phospholipase C simultaneously. PMID- 2553706 TI - Spectroscopic characterization of the iron-sulfur cluster in Bacillus subtilis glutamine phosphoribosylpyrophosphate amidotransferase. AB - The properties of the [4Fe-4S] cluster in glutamine phosphoribosylpyrophosphate amidotransferase from Bacillus subtilis have been investigated using low temperature magnetic circular dichroism, electron paramagnetic resonance (EPR), and resonance Raman spectroscopies. The Raman spectra of the native enzyme in the Fe-S stretching region show a [4Fe-4S]2+ cluster that is structurally very similar to those in simple redox proteins. Photochemical reduction mediated by 5 deazaflavin with oxalate as the electron donor resulted in [4Fe-4S]+ clusters with a mixture of ground state spin multiplicities. Magnetic circular dichroism and EPR studies of samples ranging in concentration from 0.15 to 0.4 mM concur in finding S = 3/2 [4Fe-4S]+ clusters with predominantly axial and positive zero field splitting as the dominant species. The EPR studies also revealed minor contributions from S = 1/2 [4Fe-4S]+ centers and an S = 5/2 species. The latter becomes the dominant component in more concentrated samples (approximately 2 mM), and arguments are presented in favor of assignment to S = 5/2 [4Fe-4S]+ clusters rather than adventitiously bound high spin Fe(III) ions. The concentration dependent spin state heterogeneity of the [4Fe-4S]+ cluster in glutamine phosphoribosylpyrophosphate amidotransferase is discussed in light of the magnetic and electronic properties of the [4Fe-4S]+ centers in other enzymes and proteins. PMID- 2553707 TI - An insulin-stimulated ribosomal protein S6 kinase from rabbit liver. AB - In this report we describe an activated form of S6 protein kinase in rabbits treated acutely with insulin. The major insulin-stimulated activity in rabbit liver is increased 2- to 5-fold compared to material from untreated animals based on DEAE-cellulose profiles. The activity observed in DEAE-cellulose fractions can be separated into a major and a minor peak, each having very similar chromatographic behavior. Chromatography on DEAE-cellulose, S-Sepharose, heptyl Sepharose, heparin-agarose, and Mono Q results in greater than 20,000-fold purification of the insulin-stimulated enzyme with a 12% recovery. The stimulated activity has chromatographic properties similar to an S6 protein kinase studied previously in 3T3-L1 cells (Cobb, M. H. (1986) J. Biol. Chem. 261, 12994-12999) and other systems. The enzyme purified from insulin-treated animals contains a major band that migrates in sodium dodecyl sulfate-polyacrylamide gels with Mr congruent to 70,000; this band also appears in the control preparation. Treatment of the insulin-stimulated S6 kinase with the catalytic subunit of phosphatase 2a reduces its activity by 97%. The activity of the inactivated S6 kinase is stimulated nearly 5-fold by a 15-min preincubation with partially purified insulin-stimulated microtubule-associated protein-2 kinase. PMID- 2553708 TI - Dependence of mitochondrial coenzyme A uptake on the membrane electrical gradient. AB - Coenzyme A (CoA) transport was studied in isolated rat heart mitochondria. Uptake of CoA was assayed by determining [3H]CoA associated with mitochondria under various conditions. Various oxidizable substrates including alpha-ketoglutarate, succinate, or malate stimulated CoA uptake. The membrane proton (delta pH) and electrical (delta psi) gradients, which dissipated with time in the absence of substrate, were maintained at their initial levels throughout the incubation in the presence of substrate. Addition of phosphate caused a concentration-dependent decrease of both delta pH and CoA uptake. Nigericin also dissipated the proton gradient and prevented CoA uptake. Valinomycin also prevented CoA uptake into mitochondria. Although the proton gradient was unaffected, the electrical gradient was completely abolished in the presence of valinomycin. Addition of 5 mM phosphate 10 min after the start of incubation prevented further uptake of CoA into mitochondria. A rapid dissipation of the proton gradient upon addition of phosphate was observed. Addition of nigericin or valinomycin 10 min after the start of incubation also resulted in no further uptake of CoA into with mitochondria; valinomycin caused an apparent efflux of CoA from mitochondria. Uptake was found to be sensitive to external pH displaying a pH optimum at pHext 8.0. Although nigericin significantly inhibited CoA uptake over the pHext range of 6.75-8, maximal transport was observed around pHext 8.0-8.25. Valinomycin, on the other hand, abolished transport over the entire pH range. The results suggest that mitochondrial CoA transport is determined by the membrane electrical gradient. The apparent dependence of CoA uptake on an intact membrane pH gradient is probably the result of modulation of CoA transport by matrix pH. PMID- 2553709 TI - Photoaffinity labeling of the nuclear Ah receptor from mouse Hepa 1c1c7 cells using 2,3,7,8-[3H]tetrachlorodibenzo-p-dioxin. AB - The photoinduced formation of the covalently labeled cytosolic and nuclear aryl hydrocarbon (Ah) receptors was studied using 2,3,7,8-[3H]tetrachlorodibenzo-p dioxin (TCDD) as the photoaffinity label. Irradiation of TCDD alone at wavelengths of greater than 300 nm resulted in rapid degradation of this compound (t 1/2 = 8 min). In a separate experiment, the unliganded cytosolic Ah receptor was only slowly inactivated (t 1/2 = 48 min) using the greater than 300 nm light source. Preliminary experiments with rat hepatic cytosol did not result in significant formation of specifically bound [3H]TCDD-protein covalent adducts which could be visualized by autoradiography. Irradiation of [3H]TCDD-nuclear Ah receptor complexes isolated from mouse Hepa 1c1c7 cells for 15 min gave approximately a 40% overall yield of the radiolabeled Ah receptor protein adduct. Denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the [3H]TCDD-nuclear Ah receptor photoadduct gave a single major radiolabeled protein with an apparent molecular size of 91 kDa. The chromatographic properties of the control (dark) and photolabeled nuclear Ah receptor complexes were comparable using Sephacryl S-300 and DNA-Sepharose columns. Velocity sedimentation of both the control (dark) and irradiated nuclear Ah receptor complexes gave specifically bound peaks which sedimented at 6.5 S. However, the trichloroacetic acid precipitable (buffer-reconstituted) [3H]TCDD-nuclear Ah receptor photo-covalent adduct was eluted from the Sephacryl S-300 column in the void volume and did not exhibit a specifically bound peak after velocity sedimentation analysis due to protein aggregate formation. In contrast, the elution profile of the aggregate on a DNA-Sepharose column was similar to that observed for the control (dark) and photolabeled complexes, which were eluted from the column with salt concentrations between 0.24 and 0.28 M. These photolabeling studies show that [3H] TCDD can act as a photoaffinity label for the Ah receptor and can be utilized as photoligand to probe further the structure and function of this protein. PMID- 2553710 TI - Mass changes in inositol tetrakis- and pentakisphosphate isomers induced by chemotactic peptide stimulation in HL-60 cells. AB - Absolute concentrations of inositol phosphate isomers (InsP(s] were quantified in the myeloid cell line HL-60 using the metal-dye detection technique. Stimulation with the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (fMLP) led to distinct alterations in at least seven different inositol phosphate species. Whereas the intracellular concentrations of the tetrakisphosphate isomers (InsP4(s] were found below the micromolar range, inositol 1,3,4,5,6-pentakis- and hexakisphosphate levels were about two orders of magnitude higher (36 and 54 +/- 2 microM (mean +/- S.D.), respectively). The three InsP4(s) showed distinct kinetic pattern upon receptor activation, the transient elevation of inositol 1,3,4,5-tetrakisphosphate being faster both in onset and in redecrease than inositol 1,3,4,6-tetrakisphosphate. Whereas the two latter isomers reached maximally 2.75 and 2.9 +/- 0.2 microM, respectively, 1 min after stimulation, inositol 3,4,5,6-tetrakisphosphate remained elevated (3.5 +/- 0.4 microM) up to 5 min after fMLP. Unexpected changes in highly phosphorylated InsP(s) were observed, notably a rise in inositol 1,3,4,5,6-pentakisphosphate and in inositol hexakisphosphate to 52 +/- 3 and 60 +/- 1 microM, respectively. In terms of mass, the increases in highly phosphorylated inositols are by far highest among all InsP(s). Combining radiotracer method with mass determination it was observed that the specific radioactivity of various InsP(s) was different and changed markedly upon fMLP stimulation, in spite of a prolonged labeling period leading to apparent isotopic steady state. The data presented demonstrate agonist-induced elevations of highly phosphorylated InsP(s) and suggest that inositol 1,4,5 trisphosphate, product of receptor-activated phospholipase C, is metabolized rather via phosphorylation than only by dephosphorylation pathways. PMID- 2553711 TI - The effect of endogenous essential and nonessential fatty acids on the uptake and subsequent agonist-induced release of arachidonate. AB - We have demonstrated that the uptake and agonist-induced release of a pulse of arachidonate are influenced by the size and composition of preexisting endogenous fatty acid pools. EFD-1 cells, an essential fatty acid-deficient mouse fibrosarcoma cell line, were incubated with radiolabeled (14C or 3H] arachidonate, linoleate, eicosapentaenoate (EPA), palmitate, or oleate in concentrations of 0-33 microM for 24 h. After 24 h, the cells were pulsed with 0.67 microM radiolabeled (3H or 14C, opposite first label) arachidonate for 15 min and then stimulated with 10 microM bradykinin for 4 min. Because EFD-1 cells contain no endogenous essential fatty acids, we were able to create essential fatty acid-repleted cells for which the specific activity of the newly constructed endogenous essential fatty acid pool was known. Loading the endogenous pool with the essential fatty acids arachidonate, eicosapentaenoate, or linoleate (15-20 nmol of fatty acid incorporated/10(6) cells) decreased the uptake of a pulse of arachidonate from 200 to 100 pmol/10(6) cells but had no effect on palmitate uptake. The percent of arachidonate incorporated during the pulse which was released upon agonist stimulation increased 2-fold (4-8%) as the endogenous pool of essential fatty acids was increased from 0 to 15-20 nmol/10(6) cells. This 8% release was at least 3-fold greater than the percent release from the various endogenous essential fatty acid pools. In contrast, loading the endogenous pool with the nonessential fatty acids oleate or palmitate to more than 2-3 times their preexisting cellular level had no effect on the uptake of an arachidonate pulse. Like the essential fatty acids, increasing endogenous oleate increased (by 2-fold) the percent release of arachidonate incorporated during the pulse, whereas endogenous palmitate had no effect on subsequent agonist-induced release from this arachidonate pool. These studies show that preexisting pools of essential and nonessential fatty acids exert different effects on the uptake and subsequent releasability of a pulse of arachidonate. PMID- 2553712 TI - The substitution of calcium for magnesium in H+,K+-ATPase catalytic cycle. Evidence for two actions of divalent cations. AB - In order to determine the role of divalent cations in the reaction mechanism of the H+,K+-ATPase, we have substituted calcium for magnesium, which is required by the H+,K+-ATPase for phosphorylation from ATP and from PO4. Calcium was chosen over other divalent cations assayed (barium and manganese) because in the absence of magnesium, calcium activated ATP hydrolysis, generated sufficiently high levels of phosphoenzyme (573 +/- 51 pmol.mg-1) from [gamma-32P]ATP to study dephosphorylation, and inhibited K+-stimulated ATP hydrolysis. The Ca2+-ATPase activity of the H+,K+-ATPase was 40% of the basal Mg2+-ATPase activity. However, the Ca2+,K+-ATPase activity (minus the Ca2+ basal activity) was only 0.7% of the Mg2+,K+-ATPase, indicating that calcium could partially substitute for Mg2+ in activating ATP hydrolysis but not in K+ stimulation of ATP hydrolysis. Approximately 0.1 mM calcium inhibited 50% of the Mg2+-ATPase or Mg2+,K+-ATPase activities. Inhibition of Mg2+,K+-ATPase activity was not competitive with respect to K+. Inhibition by calcium of Mg2+,K+ activity p-nitrophenyl phosphatase activity was competitive with respect to Mg2+ with an apparent Ki of 0.27 mM. Proton transport measured by acridine orange uptake was not detected in the presence of Ca2+ and K+. In the presence of Mg2+ and K+, Ca2+ inhibited proton transport with an apparent affinity similar to the inhibition of the Mg2+, K+-ATPase activity. The site of calcium inhibition was on the exterior of the vesicle. These results suggest that calcium activates basal turnover and inhibits K+ stimulation of the H+,K+-ATPase by binding at a cytosolic divalent cation site. The pseudo-first order rate constant for phosphoenzyme formation from 5 microM [gamma-32P]ATP was at least 22 times slower in the presence of calcium (0.015 s-1) than magnesium (greater than 0.310 s-1). The Ca.EP (phosphoenzyme formed in the presence of Ca2+) formed dephosphorylated four to five times more slowly that the Mg.EP (phosphoenzyme formed in the presence of Mg2+) in the presence of 8 mm trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid (CDTA) or 250 microM ATP. Approximately 10% of the Ca.EP formed was sensitive to a 100 mM KCl chase compared with greater than 85% of the Mg.EP. By comparing the transient kinetics of the phosphoenzyme formed in the presence of magnesium (Mg.EP) and calcium (Ca.EP), we found two actions of divalent cations on dephosphorylation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2553713 TI - cDNA cloning, functional expression, and mRNA tissue distribution of a third organellar Ca2+ pump. AB - We describe the characterization of a rat kidney cDNA that encodes a novel Ca2+ transporting ATPase. The cDNA, termed RK 8-13, was isolated previously using an oligonucleotide hybridization probe corresponding to part of the ATP binding site of the sarcoplasmic reticulum Ca-ATPases (Gunteski-Hamblin, A.-M., Greeb, J., and Shull, G. E. (1988) J. Biol. Chem. 263, 15032-15040). The complete nucleotide sequence of the 4.5-kilobase cDNA has been determined, and the primary structure of the protein has been deduced. The enzyme consists of 999 amino acids, has an Mr of 109,223, and contains all of the conserved domains found in transport ATPases of the E1-E2 class. It exhibits 75-77% amino acid identity with the fast twitch and slow-twitch/cardiac isoforms of the sarcoplasmic reticulum Ca-ATPase, and the hydropathy plots of the three enzymes are virtually identical. High levels of ATP-dependent Ca2+ uptake were demonstrated in microsomes of COS-1 cells that had been transfected with a construct consisting of the entire coding sequence of the cDNA in the expression vector p91023(B). Northern blot analyses of poly(A)+ RNA revealed that the mRNA for this protein is expressed in heart, skeletal muscle, uterus, brain, lung, liver, kidney, testes, small intestine, large intestine, and pancreas. These data show that the enzyme is a Ca2+ transporting ATPase and that its mRNA is expressed in a broad variety of both muscle and non-muscle tissues. PMID- 2553714 TI - A high concentration of SecA allows proton motive force-independent translocation of a model secretory protein into Escherichia coli membrane vesicles. AB - The in vitro translocation of OmpF-Lpp, a model secretory protein, into inverted membrane vesicles of Escherichia coli obligatorily requires the proton motive force (delta mu H+) in the conventional assay system (Yamada, H., Tokuda, H., and Mizushima, S. (1989) J. Biol. Chem. 264, 1723-1728). The translocation, however, took place efficiently, even in the absence of delta mu H+, when the system was supplemented with additional SecA. With the stripped membrane vesicles, which are permeable to protons, or in the absence of NADH, the supplementation of SecA remarkably stimulated the translocation activity. The further addition of NADH did not significantly enhance the translocation activity under the SecA-enriched conditions. OmpF-Lpp thus translocated could be recovered from the vesicular lumen by sonication, indicating that complete translocation occurred in the absence of delta mu H+. It is suggested that delta mu H+ is required for high affinity interaction of SecA with the presumed secretory machinery in the cytoplasmic membrane and that a high concentration of SecA modulates the delta mu H+ requirement. PMID- 2553715 TI - In vitro analysis of the process of translocation of OmpA across the Escherichia coli cytoplasmic membrane. A translocation intermediate accumulates transiently in the absence of the proton motive force. AB - The proton motive force (delta mu H+) plays an important role, although it is not absolutely essential, in the in vitro translocation of secretory proteins, such as OmpA, across the cytoplasmic membrane of Escherichia coli (Yamada, H., Tokuda, H., and Mizushima, S. (1989) J. Biol. Chem. 264, 1723-1728). The transient accumulation in membrane vesicles of a possible translocation intermediate of OmpA was observed in the absence of delta mu H+. The intermediate was detected on a polyacrylamide gel as a proteinase K-resistant band corresponding to a molecular weight of 26,000. The intermediate did not possess the signal peptide. The appearance of this band was inhibited in the absence of ATP or the presence of adenosine 5'-(beta,gamma-imino)triphosphate (AMP-PNP) and enhanced upon the addition of SecA. Upon the addition of NADH that energizes the membrane, the intermediate was converted to the translocated form of OmpA, even in the presence of AMP-PNP. These results suggest different requirements of ATP and delta mu H+ for the early and late stages of the translocation reaction. The SecA requirement for the early stage of the translocation has also been suggested. In addition to this band, two other bands were observed at higher positions on the gel, when the translocation reaction was performed in the absence of delta mu H+. Although these two bands also represented the mature form of OmpA, which was partly protected from the proteinase K treatment by the membrane vesicles, the accumulation was not transient. These bands did not appear when the translocation reaction was performed in the presence of dithiothreitol. Together with other evidence, the above observations suggest that OmpA, which has an intramolecular disulfide bridge, cannot undergo the translocation unless delta mu H+ is imposed. PMID- 2553716 TI - Aldosterone induction of electrogenic sodium transport in the apical membrane vesicles of rat distal colon. AB - Na-H exchange is present in apical membrane vesicles (AMV) isolated from distal colon of normal rats. Because in intact tissue aldosterone both induces amiloride sensitive electrogenic sodium transport and inhibits electroneutral sodium absorption, these studies with AMV were designed to establish the effect of aldosterone on sodium transport. An outward-directed proton gradient stimulated 22Na uptake in AMV isolated from distal colon of normal and dietary sodium depleted (with elevated aldosterone levels) experimental rats. Unlike normal AMV, proton gradient-dependent 22Na uptake in experimental AMV was inhibited when uptake was measured under voltage-clamped conditions. 10 microM amiloride inhibited the initial rate of proton gradient-dependent 22Na uptake in AMV of normal and experimental rats by 30 and 75%, respectively. In contrast, 1 mM amiloride produced comparable inhibition (90 and 80%) of 22Na uptake in normal and experimental AMV. Intravesicular-negative potential stimulated 22Na uptake in experimental but not in normal AMV. This increase was inhibited by 90% by 10 microM amiloride. An analogue of amiloride, 5-(N-ethylisopropyl) amiloride (1 microM), a potent inhibitor of electroneutral Na-H exchange in AMV of normal rat distal colon, did not alter potassium diffusion potential-dependent 22Na uptake. Increasing sodium concentration saturated proton gradient-dependent 22Na uptake in normal AMV. However, in experimental AMV, 22Na uptake stimulated by both proton gradient and potassium diffusion potential did not saturate as a function of increasing sodium concentration. We conclude from these results that an electrically sensitive conductive channel, not electroneutral Na-H exchange, mediates 22Na uptake in AMV isolated from the distal colon of aldosterone rats. PMID- 2553717 TI - Isolation of a receptor for acidic and basic fibroblast growth factor from embryonic chick. AB - A receptor for acidic and basic fibroblast growth factors (aFGF and bFGF, respectively) was isolated from 7-day embryonic chick. Chromatography of solubilized membrane proteins on wheat germ agglutininagarose and aFGF-Sepharose yielded three major polypeptides migrating at 150, 70, and 45 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These polypeptides were eluted from aFGF-Sepharose with either 1.0 M NaCl or 100 micrograms/ml heparin, but were not retained on underivatized Sepharose. Cross-linking of 125I aFGF or 125I-bFGF to either crude membrane preparations or to purified fractions yielded a 165-kDa complex, suggesting the existence of a 150-kDa FGF receptor after subtraction of approximately 15 kDa for 125I-FGF. Addition of excess aFGF or bFGF competed for binding of either 125I-aFGF or 125I-bFGF to FGF receptor preparations. Purified FGF receptor fractions were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to Immobilon membranes, and incubated with 125I-aFGF or 125I-bFGF in order to identify FGF-binding polypeptides. Bound 125I-aFGF and 125I-bFGF were displaced by aFGF and bFGF, but not epidermal growth factor, consistent with the identification of the 150-kDa polypeptide as a receptor for acidic and basic FGF. Treatment of purified FGF receptor fractions with N-glycanase demonstrated that the 150-kDa polypeptide contained approximately 10 kDa of N-linked oligosaccharide. The apparent molecular mass of the 150-kDa polypeptide was unaffected by treatment with heparitinase, indicating that the 150-kDa polypeptide is not a heparan sulfate proteoglycan. Together, these data suggest that the 150-kDa polypeptide is a FGF receptor that may mediate the biological activities of aFGF and bFGF. PMID- 2553718 TI - Affinity purification of the C alpha and C beta isoforms of the catalytic subunit of cAMP-dependent protein kinase. AB - A synthetic peptide of 18 amino acids corresponding to the inhibitory domain of the heat-stable protein kinase inhibitor was synthesized and shown to inhibit both the C alpha and C beta isoforms of the catalytic (C) subunit of cAMP dependent protein kinase. Extracts from cells transfected with expression vectors coding for the C alpha or the C beta isoform of the C subunit required 200 nM protein kinase inhibitor peptide for half-maximal inhibition of kinase activity in extracts from these cells. An affinity column was constructed using this synthetic peptide, and the column was incubated with protein extracts from cells overexpressing C alpha or C beta. Elution of the affinity column with arginine allowed single step isolation of purified C alpha and C beta subunits. The C alpha and C beta proteins were enriched 200-400-fold from cellular extracts by this single step of affinity chromatography. No residual inhibitory peptide activity could be detected in the purified protein. The purified C subunit isoforms were used to demonstrate preferential antibody reactivity with the C alpha isoform by Western blot analysis. Furthermore, preliminary characterization showed both isoforms have similar apparent Km values for ATP (4 microM) and for Kemptide (5.6 microM). These results demonstrate that a combination of affinity chromatography employing peptides derived from the heat-stable protein kinase inhibitor protein and the use of cells overexpressing C subunit related proteins may be an effective means for purification and characterization of the C subunit isoforms. Furthermore, this method of purification may be applicable to other kinases which are known to be specifically inhibited by small peptides. PMID- 2553719 TI - Poly(ADP-ribose)-mediated post-translational modification of chromatin-associated human topoisomerase I. Inhibitory effects on catalytic activity. AB - We have investigated the association of human topoisomerase I with poly(ADP ribosylated) domains of chromatin and the effects of this modification on the enzyme activity. In vitro poly(ADP-ribosylation) assays demonstrated that this enzyme was one of the major acceptors for this chromatin-dependent post translational modification. Western blotting procedures using antibody to topoisomerase I indicated that under extensive poly(ADP-ribosylation) conditions, where a majority of poly(ADP-ribose) acceptor molecules form aggregates, the major population of the topoisomerase I associated with chromatin was apparently non-aggregated. The catalytic activity of the topoisomerase I associated with the poly(ADP-ribosylated) chromatin was 3-5-fold inhibited. Additionally, antibody to poly(ADP-ribose) was used to immunofractionate selectively the modified domains of chromatin. Our data suggests the presence of topoisomerase I, both adjacent and distal to the poly(ADP-ribosylated) sites of chromatin. Unmodified and a significant portion of the modified species of enzyme migrated as approximately 100-kDa proteins. However, the modified form of topoisomerase was noted to be catalytically less active as compared to the enzyme bound to the non-poly(ADP ribosylated) nucleosomes. These results provide evidence, at the cellular level, for the poly(ADP-ribosylation)-mediated regulation of human topoisomerase I and suggest a functional significance for poly(ADP-ribosylation) in topoisomerase related processes (replication, transcription, and recombination) in eukaryotes. PMID- 2553720 TI - Characterization of OmpR binding sequences in the upstream region of the ompF promoter essential for transcriptional activation. AB - In Escherichia coli the expression of the outer membrane porin gene ompF requires the transcriptional activator protein OmpR. Previous DNase I footprinting experiments with purified OmpR localized the OmpR binding site from positions 105 to -60 (relative to the transcriptional start site) in the ompF promoter, and three tandem 10-base pair sequences elements (Fa, Fb, and Fc) within this region were proposed to be important for OmpR recognition. In order to elucidate the roles of the F boxes for transcriptional activation of ompF, various F box deletions and point mutations were constructed and analyzed for their effects on ompF-lacZ expression and OmpR binding. Removal of 102 nucleotides, which included a portion of the OmpR binding region (the Fa box), evidenced the largest decrease in transcriptional activation and significantly reduced OmpR binding. Additional deletion of four more base pairs in this target site (representing half of the Fb box) further reduced ompF expression. OmpR interactions with DNA sequences representing the OmpR binding region were analyzed by DNA mobility shift experiments. A 43-base pair ompF oligonucleotide containing the Fa, Fb, and Fc regions was sufficient for OmpR-dependent DNA binding using either purified OmpR or cell supernatants. The central C residue in each F box was changed to a T and unique patterns of protein-DNA complexes were observed that were different from that of the wild type binding site. The most dramatic effect on OmpR binding was observed when the C to T transversion occurred in the Fb box, and this mutation also reduced the level of ompF-lacZ expression. Our results indicate that the F boxes play important roles in the activation of ompF expression, and we suggest that OmpR may interact cooperatively with these boxes. PMID- 2553721 TI - Identification of the carboxyl-terminal amino acids important for the ADP ribosylation activity of Pseudomonas exotoxin A. AB - The ADP-ribosylation domain of Pseudomonas exotoxin A (PE) has been identified to reside in structural domain III (residues 405-613) and a portion of domain Ib (residues 385-404) of the molecule (Hwang, J., FitzGerald, D. J., Adhya, S., and Pastan, I. (1987) Cell 48, 129-136). To further determine the carboxyl end region essential for ADP-ribosylation activity, we constructed sequential deletions at the carboxyl-terminal of PE. Our results show that a clone with a deletion of the carboxyl-terminal amino acid residues from Arg-609 to Lys-613 and replaced with Arg-Asn retained wild-type PE ADP-ribosylation activity. Deletion of the terminal amino acid residues from Ala-596 to Lys-613 and replaced with Val-Ile-Asn reduced ADP-ribosylation activity by 75%, while deletions of 36 or more amino acids from the carboxyl terminus completely lose their ADP-ribosylation activity. These modified PEs were also examined for their ability to block PE cytotoxicity. Our results shown that modified PEs which lost their ADP-ribosylation activity correspondingly lost their cytotoxicity. Furthermore, extracts containing PE fragments without ADP-ribosylation activity were able to block the cytotoxic activity of intact PE. Our results thus indicate that carboxyl-terminal amino acids in the Ser-595 region are crucial for ADP-ribosylation activity and, consequently, cytotoxicity of PE. The modified PEs which have lost their ADP ribosylation activity may also be a route to new PE vaccines. PMID- 2553722 TI - Purification and characterization of the lipoprotein-associated coagulation inhibitor from human plasma. AB - The lipoprotein-associated coagulation inhibitor (LACI) has been isolated from human plasma using a combination of hydrophobic, ion-exchange, and affinity chromatography. The final purification required was greater than 500,000-fold with a yield of 13%. Plasma LACI, on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, contains major bands at 40 and 46 kDa and minor bands at 55, 65, 75, 90, and approximately 130 kDa. All of the molecular weight forms are recognized by antibodies to LACI's amino and carboxyl termini and are able to inhibit the factor VII(a)-tissue factor complex and factor Xa. Plasma LACI, reduced with beta-mercaptoethanol, migrates on sodium dodecyl-sulfate polyacrylamide gel electrophoresis as a doublet at 42 kDa and has an amino terminal sequence essentially identical to that of HepG2 LACI. The difference in size between reduced plasma LACI (42 kDa) and HepG2 LACI (47 kDa) may be related to differing degrees of N-linked glycosylation. The 46-kDa and larger forms of unreduced plasma LACI are associated with apolipoprotein A-II (apoA-II) in mixed disulfide linkages. Studies using isolated lipoproteins show that low density lipoprotein (LDL) contains primarily the 40-kDa form of LACI, whereas high density lipoprotein (HDL) contains primarily the 46-kDa form of LACI (LACI/apoA II complexes). Gel filtration of a fresh plasma sample showed approximately 50% of plasma LACI to be associated with LDL/very low density lipoprotein, 44% with HDL, and the remaining 6% to not be associated with lipoproteins. PMID- 2553723 TI - Properties of erythrocyte membrane binding and autolytic activation of calcium activated neutral protease. AB - The binding of a calcium-activated neutral protease (CANP) with high calcium sensitivity (muCANP) to erythrocyte membranes and its subsequent autolytic activation on the membranes were analyzed by an immunoblot technique. In the presence of calcium ions, muCANP bound to the erythrocyte membranes as a heterodimer of 79- and 28-kDa subunits and was converted quickly on the membranes to an active form with a 76-kDa large subunit. The active form was then released from the membranes to the soluble fraction. These sequential reactions, however, were not specific to inside-out vesicles, but occurred also, except for some Ca2+ independent binding, on right side-out vesicles. A rapid degradation of some membrane proteins was observed after binding of muCANP to the membranes. The binding of muCANP to erythrocyte membranes was inhibited by substrates and the endogenous CANP inhibitor, which is also a suicide substrate. These results strongly suggest that muCANP binds to membranes by recognition of membrane proteins as substrates and not at a special site for activation. Thus, a possible mechanism for muCANP activation on membranes is that muCANP first binds to substrates on membranes, is activated, and then degrades the substrates to deform the membrane structures. PMID- 2553724 TI - Inhibition of calpain by a synthetic oligopeptide corresponding to an exon of the human calpastatin gene. AB - Calpastatin is a widely distributed endogenous inhibitor protein specifically acting on calpain (Ca2+-dependent cysteine endopeptidase). The inhibitor consists of four inhibitory domains (Domains 1-4) with mutually homologous sequences. NH2 terminal Domain L is non-homologous, and all domains have 120-140 residues each. A human calpastatin genomic DNA clone was isolated using a previously obtained human calpastatin cDNA probe. Sequence analysis has revealed that the clone contains Domain 1 and segments of neighboring domains (Domains L and 2). Each of three highly conserved, restricted regions within Domain 1 was located on separate exons, 1A, 1B, and 1C. Exon 2A, corresponding to the first exon of Domain 2, is homologous to Exon 1A and follows Exon 1D of Domain 1. A 27-residue peptide encoded by Exon 1B, including a 12-residue middle conserved sequence, was chemically synthesized and tested for protease inhibitory activities. The synthetic peptide showed strong inhibition against calpain I (low Ca2+-requiring form), and calpain II (high Ca2+-requiring form), but no inhibition against papain or trypsin. These results indicated that Exon 1B forms a self-sufficient functional subdomain of the calpastatin inhibitory domain. PMID- 2553725 TI - Functional expression of the human HepG2 and rat adipocyte glucose transporters in Xenopus oocytes. Comparison of kinetic parameters. AB - Facilitated glucose transport is a ubiquitous characteristic of animal cells carried out by a family of membrane glycoproteins. Two members of this gene family are the well characterized human erythrocyte protein that has been cloned from the HepG2 cell line and the insulin-sensitive transporter that has been cloned from adipocytes and muscle tissue. In the present study the HepG2 and adipocyte glucose transporters were functionally expressed in Xenopus oocytes after injection of synthetic mRNAs produced by transcription in vitro from cloned cDNAs. Both 2-deoxyglucose uptake and 3-O-methylglucose transport were increased several-fold over basal levels in mRNA-injected oocytes. Increased uptake of 2 deoxyglucose was inhibited completely in the presence of cytochalasin B, and 3-O methylglucose transport was blocked by D-glucose but not by L-glucose. The half saturation constant and turnover number for 3-O-methylglucose transport at 22 degrees C via the HepG2 transporter were estimated to be 21 mM and 2.2 x 10(3) s 1 under equilibrium exchange conditions. The half-saturation constant for 3-O methylglucose transport via the adipocyte transporter under the same conditions was estimated to be 1.8 mM. These data prove the functional identity of the cloned HepG2 and adipocyte cDNAs and indicate that the HepG2 and adipocyte transporters display similar kinetic behavior when expressed in the frog oocyte membrane as compared with their native membrane environments. Thus, the difference in the equilibrium exchange kinetic parameters for glucose transport in the erythrocyte and the adipocyte is a result of the expression of two distinct glucose transporter proteins. PMID- 2553726 TI - Measurement and characterization of postischemic free radical generation in the isolated perfused heart. AB - Electron paramagnetic resonance spectroscopy has been applied to measure radical generation in the postischemic heart; however, there is controversy regarding the methods used and the conclusion as to whether radicals are generated. In order to resolve this controversy, direct and spin trapping measurements of the time course and mechanisms of radical generation were performed in isolated perfused rabbit hearts. In reperfused tissue, 3 prominent radical signals are observed: A, isotropic g = 2.004 suggestive of a semiquinone; B, anisotropic g parallel = 2.033 and g perpendicular = 2.005 suggestive of ROO.; and C, a triplet g = 2.000 and aN = 24 G suggestive of a nitrogen centered radical. B and C, however, are highly labile and disappear at temperatures probably encountered in some previous studies. In normally perfused hearts, A is observed with only small amounts of B and C. During ischemia, B and C increase reaching a maximum after 45 min while A decreases. On reflow with oxygenated perfusate all 3 signals increase. With varying duration of ischemia and reflow, peak signal intensities occurred after 15 s of reflow following 30 min of ischemia. Reperfusion with superoxide dismutase, deferoxamine, or mannitol abolished the reperfusion increase of B. Measurements performed with the spin trap 5,5'-dimethyl-1-pyrroline-N-oxide (DMPO) demonstrated a similar time course of radical generation with prominent DMPO-OH and DMPO-R signals peaking between 10 and 20 s of reflow. Superoxide dismutase and deferoxamine also quenched these signals. Thus, .O2- derived .OH, R., and ROO. radicals are generated in postischemic myocardium. While the experimental techniques used can result in loss of intrinsic radicals and generation of extraneous radicals, with proper care and controls valid measurements of free radicals in biological tissues can be performed. PMID- 2553727 TI - Monoclonal antibodies mimic insulin activation of ribosomal protein S6 kinase without activation of insulin receptor tyrosine kinase. Studies in cells transfected with normal and mutant human insulin receptors. AB - The effects of species-specific monoclonal antibodies to the human insulin receptor on ribosomal protein S6 phosphorylation were studied in rodent cell lines transfected with human insulin receptors. First, Swiss mouse 3T3 fibroblasts expressing normal human insulin receptors (3T3/HIR cells) were studied. Three monoclonal antibodies, MA-5, MA-20, and MA-51, activated S6 kinase in these cells but had no effects in untransfected 3T3 cells. Both insulin and MA 5, the most potent antibody, activated S6 kinase in a similar time- and dose dependent manner. To measure S6 phosphorylation in vivo, 3T3/HIR cells were preincubated with [32P]Pi and treated with insulin and MA-5. Both agents increased S6 phosphorylation, and their tryptic phosphopeptide maps were similar. MA-5 and the other monoclonal antibodies, unlike insulin, failed to stimulate insulin receptor tyrosine kinase activity either in vitro or in vivo. Moreover, unlike insulin, they failed to increase the tyrosine phosphorylation of the endogenous cytoplasmic protein, pp 185. Next, HTC rat hepatoma cells, expressing a human insulin receptor mutant that had three key tyrosine autophosphorylation sites in the beta-subunit changed to phenylalanines (HTC-IR-F3 cells), were studied. In this cell line but not in untransfected HTC cells, monoclonal antibodies activated S6 kinase without stimulating either insulin receptor autophosphorylation or the tyrosine phosphorylation of pp 185. These data indicate, therefore, that monoclonal antibodies can activate S6 kinase and then increase S6 phosphorylation. Moreover, they suggest that activation of receptor tyrosine kinase and subsequent tyrosine phosphorylation of cellular proteins may not be crucial for activation of S6 kinase by the insulin receptor. PMID- 2553728 TI - Human bisphosphoglycerate mutase. Expression in Escherichia coli and use of site directed mutagenesis in the evaluation of the role of the carboxyl-terminal region in the enzymatic mechanism. AB - Bisphosphoglycerate mutase is an erythrocyte-specific enzyme whose main function is to synthesize 2,3-diphosphoglycerate, the allosteric effector of hemoglobin. In addition to its main 2,3-diphosphoglycerate synthase activity, the enzyme displays phosphatase and mutase activities both involving 2,3-diphosphoglycerate in their reaction. The three activities have been demonstrated to be catalysed at a unique active site. To study the structure of such an active site we have developed a recombinant system producing mutants of human bisphosphoglycerate mutase in Escherichia coli, by site-directed mutagenesis. For this purpose the human bisphosphoglycerate mutase cDNA that we had previously cloned has been used to construct a procaryotic high level expression vector bearing the "tac" promoter. Human bisphosphoglycerate mutase produced in E. coli, a species which does not normally synthesize this enzyme, represented 8% of the total soluble bacterial protein and displayed the three catalytic activities (synthase, mutase, and phosphatase) characteristic of the enzyme. Since it has been suggested that the carboxyl-terminal region may be implicated in the catalytic activity of the enzyme, three variants deleted in this part of the protein were produced. Our results indicate that a minimal deletion of 7 amino acid residues in the carboxyl terminal portion of the human bisphosphoglycerate mutase completely abolished the three catalytic activities of the enzyme. In contrast, the effects of the deletion of the last two lysine residues were limited to a 38% reduction in the synthase activity. These results show that the carboxyl-terminal amino acid residues are either directly or indirectly implicated in the three catalytic functions of the human bisphosphoglycerate mutase, and that the two terminal lysine residues are not essential for the major part of the enzymatic mechanism of the enzyme. PMID- 2553729 TI - Ligand binding to cytochrome c peroxidase from Pseudomonas aeruginosa. AB - The high potential heme site of Pseudomonas cytochrome c peroxidase has His and Met as ligands. On reduction, the Fe-met bond becomes photosensitive. Following photolysis, the bond reforms with a half-time of 35 ps. The low potential heme peroxidatic site of the fully reduced enzyme has been shown to bind to a range of ligands. The compounds with carbon monoxide, methyl, ethyl, n-butyl, and t-butyl isonitriles have been investigated by laser flash photolysis. All are photosensitive and show different degrees of geminate recombination of ligand in the picosecond and nanosecond time ranges. Carbon monoxide shows the least effect. The three straight-chain isonitriles show about 50% geminate recombination with half-times of the order of 10 ns. t-Butyl isonitrile shows more and faster recombination. These results imply considerable freedom of movement within the active site for the smaller ligands. PMID- 2553730 TI - Herpes simplex virus type 1 and human DNA polymerase interactions with 2' deoxyguanosine 5'-triphosphate analogues. Kinetics of incorporation into DNA and induction of inhibition. AB - The ability of herpes simplex virus type 1 (HSV-1) DNA polymerase, HeLa polymerase alpha, and HeLa polymerase beta to utilize several dGTP analogues has been investigated using a defined synthetic template primer. The relative efficiencies of the triphosphates of 9-[(2-hydroxyethoxy)methyl]guanine (acyclovir triphosphate, ACVTP), 9-[(1,3-dihydroxy-2-propoxy)methyl] guanine (ganciclovir triphosphate, DHPGTP), and 2',3'-dideoxyguanosine (ddGTP) as substrates for the three polymerases were: HSV-1 polymerase, dGTP greater than ACVTP approximately equal to DHPGTP greater than ddGTP; polymerase alpha, dGTP greater than ACVTP approximately equal to DHPGTP much greater than ddGTP; polymerase beta, ddGTP greater than dGTP much greater than ACVTP approximately equal to DHPGTP. The potent inhibition of HSV-1 polymerase by ACVTP has been shown previously to be due to the formation of a dead-end complex upon binding of the next 2'-deoxynucleoside 5'-triphosphate encoded by the template after incorporation of acyclovir monophosphate into the 3' end of the primer (Reardon, J. E., and Spector, T. (1989) J. Biol. Chem. 264, 7405-7411). This mechanism was shown here to be a general mechanism for inhibition of polymerases by the obligate chain terminators, ACVTP and ddGTP. The ACVTP-induced inhibition was 30 fold more potent with HSV-1 polymerase than with polymerase alpha. This difference may contribute to the antiviral selectivity of this nucleotide analogue. The effect of ganciclovir monophosphate incorporation (a nonobligate chain terminator) on subsequent primer extension was also evaluated. With HSV-1 polymerase and polymerase alpha, although there was a considerable reduction in the efficiency of utilization of the 3'-DHPGMP-terminal primer, contrasting kinetic behavior was observed. With HSV-1 polymerase, insertion of DHPGTP resulted in a significant reduction in Vmax for subsequent nucleotide incorporations. In contrast, with polymerase alpha, a relatively small decrease in Vmax was accompanied by increased Km values for subsequent nucleotide incorporations. PMID- 2553731 TI - Expression and characterization of calmodulin-dependent protein kinase II from cloned cDNAs in Chinese hamster ovary cells. AB - cDNAs containing the entire coding regions of the alpha and beta subunits of calmodulin-dependent protein kinase II (CaM kinase II) were isolated from a rat cerebrum cDNA library, ligated into an expression vector under the control of SV40 early promoter and introduced into Chinese hamster ovary (CHO) cells. To investigate the role of the alpha and beta subunits and their functional domains in CaM kinase II activity, the properties of the kinases expressed in the transfected cells were studied. CaM kinase II activity was detected in the transfected cells when the alpha and beta cDNAs were introduced into CHO cells simultaneously. RNA transfer blot and protein immunoblot analyses demonstrated the expression of the mRNAs and proteins of both alpha and beta subunits in the cloned cells. When alpha or beta cDNA was introduced into CHO cells separately, a significant level of the enzyme activity was also expressed, indicating that the alpha and beta subunits exhibited enzyme activity individually. The apparent Km values for ATP and MAP 2 were almost the same for the alpha subunit, beta subunit, alpha beta complex, and brain CaM kinase II. However, there was a slight difference in the affinity for calmodulin between the expressed proteins. The alpha and beta subunits expressed in the same cells polymerized to form alpha beta complex of a size similar to that of brain CaM kinase II. The alpha subunit also polymerized to form an oligomer, which showed almost the same S value as that of alpha beta complex and brain CaM kinase II. In contrast, the beta subunit did not polymerize. The alpha subunit, beta subunit, alpha beta complex, and brain CaM kinase II were autophosphorylated with [gamma-32P]ATP in the presence of Ca2+ and calmodulin, which resulted in the appearance of Ca2+-independent activity. The Ca2+-independent activity was 60-75% of the total activity as measured in the presence of Ca2+ plus calmodulin. To examine the functional relationship of peptide domains of the subunits of CaM kinase II, deleted cDNAs were introduced into CHO cells and the properties of the expressed proteins were studied. In cells transfected with alpha or beta cDNA from which the association domain was deleted, a significant level of kinase activity was expressed. However, the expressed proteins showed hardly any autophosphorylation and the appearance of Ca2+-independent enzyme activity was very low, indicating that the association domain was essential for the autophosphorylation and for the appearance of the Ca2+-independent activity. PMID- 2553732 TI - Purification and characterization of an insulin-like growth factor II variant from human plasma. AB - An insulin-like growth factor II variant (IGF-II variant) was purified from Cohn fraction IV1 of human plasma by ion exchange, gel filtration, and reversed-phase high pressure liquid chromatography. The amino-terminal sequence of the first 35 amino acid residues showed a replacement of Ser-29 of IGF-II with the tetrapeptide Arg-Leu-Pro-Gly of IGF-II variant. Peptides isolated and sequenced after digestion with endoproteinase Asp-N and endoproteinase Glu-C disclosed no differences with the sequence predicted from an IGF-II variant cDNA clone isolated by Jansen, M., van Shaik, F. M. A., van Tol, H., Van den Brande, J. L., and Sussenbach, J. S. (1985) FEBS Lett., 179, 243-246. The molecular ion of intact IGF-II variant was 7809.4 mass units, as measured by plasma desorption mass spectrometry. This is in close agreement with the molecular ion of 7812.8 mass units calculated from the determined sequence and indicates the entire amino acid sequence had been accounted for. Binding of IGF-II variant to purified insulin-like growth factor I (IGF-I) receptors demonstrated a 2-3-fold lower affinity for this receptor compared with IGF-I or IGF-II. The dissociation constants for IGF-I, IGF-II, and IGF-II variant are 0.23, 0.38, and 0.80 nM, respectively. In a growth assay, the concentration of IGF-II and IGF-II variant required to stimulate the half-maximal growth of MCF-7 cells was 4 and 13 nM, respectively. Finally, the amount of IGF-II variant that can be purified by this method constitutes approximately 25% of the total IGF-II isolated from Cohn fraction IV1 of human plasma. PMID- 2553733 TI - Nitrogen fixation (nif) genes of the cyanobacterium Anabaena species strain PCC 7120. The nifB-fdxN-nifS-nifU operon. AB - A second nitrogen fixation (nif) operon in the cyanobacterium (blue-green alga) Anabaena (Nostoc) sp. strain PCC 7120 has been identified and sequenced. It is located just upstream of the nifHDK operon and consists of four genes in the order nifB, fdxN, nifS, and nifU. The three nif genes were identified on the basis of their similarity with the corresponding genes from other diazotrophs. The fourth gene, fdxN, codes for a bacterial type ferredoxin (Mulligan, M. E., Buikema, W. J., and Haselkorn, R. (1988) J. Bacteriol. 167, 4406-4410). The four genes are probably transcribed as a single operon, but are expressed at a lower level than the nifHDK operon, and only after a developmentally induced DNA rearrangement occurs that excises a 55-kilobase pair element from within the fdxN gene (Golden, J. W., Mulligan, M. E., and Haselkorn, R. (1987) Nature 327, 526 529; Golden, J. W., Carrasco, C. D., Mulligan, M. E., Schneider, G. J., and Haselkorn, R. (1988) J. Bacteriol. 170, 5034-5041). The promoter for the nifB operon was located by primer extension. Comparison of the nifB 5'-flanking sequence with the nifH 5'-flanking sequence did not reveal any consensus base pairs that would define a nif promoter for Anabaena. The operon contains two instances of 7-base pair directly repeated sequences: seven copies of the repeated sequence are found between the nifB and fdxN genes and six copies are found between the nifS and nifU genes. The function of these repeats is unknown. PMID- 2553734 TI - Arsenite inhibits two steps in the ubiquitin-dependent proteolytic pathway. AB - Eukaryotic cells possess a multienzyme ATP-dependent proteolytic pathway in which the small, highly conserved protein ubiquitin (Ub) acts as a cofactor. In this pathway, formation of a covalent Ub-substrate conjugate precedes ATP-dependent degradation of the substrate. Inorganic arsenite (AsO2-) inhibited Ub-dependent protein degradation in a concentration-dependent fashion, both in intact rabbit reticulocytes and in a reticulocyte lysate (fraction II). Concentrations of arsenite causing half-maximal inhibition in fraction II varied with the substrate, ranging from 0.025 mM (bovine alpha-lactalbumin) to 3.3 mM (reduced/carboxymethylated bovine serum albumin). Inhibition was rapidly reversed upon addition of dithiothreitol. Arsenite inhibited the Ub-dependent proteolytic pathway at one or both of two steps, depending on the substrate. 1) Proteins with acidic amino termini must be amino terminally arginylated, in a tRNA-dependent reaction, prior to becoming conjugated to Ub (Ferber, S., and Ciechanover, A. (1987) Nature 326, 808-811). Arsenite inhibited substrate arginylation, and therefore also inhibited Ub conjugation. The inhibited species appeared to be arginyl aminoacyl-tRNA transferase, since arsenite was without strong effect on the rate or extent of arginyl-tRNA formation in fraction II, yet almost completely inhibited arginine transfer from arginyl-tRNA to reduced/carboxymethylated bovine serum albumin. 2) Arsenite also inhibited Ub substrate conjugate turnover, as shown in pulse-chase experiments. For a given substrate, degradative (protease-dependent) and Ub regenerative (isopeptidase dependent) components of conjugate turnover were similarly inhibited by arsenite. The potency of this inhibition varied for different substrates. Monosubstituted trivalent arsenicals such as arsenite typically interact with sites containing vicinal sulfhydryl groups. Inhibition by arsenite of two steps in the Ub dependent proteolytic pathway suggests that the relevant pathway components could possess this kind of structural/catalytic feature. PMID- 2553735 TI - Herpes simplex-1 DNA polymerase. Identification of an intrinsic 5'----3' exonuclease with ribonuclease H activity. AB - The herpes simplex virus-1 DNA polymerase is a heterodimer of Mr 190,000 which consists of the products of the UL30 (Pol) and UL42 genes. The 136-kilodalton Pol gene product contains an intrinsic ribonuclease H activity that specifically degrades RNA.DNA heteroduplexes or duplex DNA substrates in the 5'----3' direction. It can therefore catalyze the excision of the RNA primers that initiate the synthesis of Okazaki fragments at a replication fork during herpes DNA replication. PMID- 2553736 TI - The brown alga Ascophyllum nodosum contains two different vanadium bromoperoxidases. AB - Haloperoxidases have been detected in a variety of organisms, including bacteria, fungi, algae, and mammals. Mammalian haloperoxidases are known to be directly involved in the oxidative destruction of microorganisms. The algal bromoperoxidases are probably involved in the biosynthesis of bromometabolites, most of which show considerable bactericidal activity. From the brown seaweed Ascophyllum nodosum (order, Fucales) two different bromoperoxidases have been isolated, which both contain vanadium as an essential element for enzymic activity. The location of these two enzymes, determined by activity staining of cross-sections of algal parts, was different. Bromoperoxidase I (which has been described before) was located inside the thallus, particularly around the conceptacles, whereas bromoperoxidase II was present at the thallus surface of the alga. The molecular masses of these bromoperoxidases as judged from sodium dodecyl sulfate-gel electrophoresis were 97 and 106 kDa, respectively. Some of the enzymatic properties (pH optimum and Km for bromide) of the two enzymes were slightly different, whereas the amino acid compositions were more or less equal. The isoelectric point of the two proteins was the same, namely 5.0. On sodium dodecyl sulfate-polyacrylamide gels both enzymes could be stained with periodic acid Schiff's reagent, so both are glycoproteins. Since only bromoperoxidase II could be bound to a concanavalin A-Sepharose column, these enzymes contain different carbohydrates. Both enzymes display a considerable thermostability. However, the chemical stability of the two bromoperoxidases differed. Bromoperoxidase II could also be inactivated by dialysis at low pH and reactivation was only possible with the transition metal vanadium and not with other metal ions. The presence of vanadium in this enzyme could be established with atomic absorption spectrophotometry and electron paramagnetic resonance. The EPR signals of both bromoperoxidases, which were observed after reduction with sodium dithionite, were similar: only minor differences were observed in the hyperfine coupling. In immunoblotting experiments these two bromoperoxidases were found to cross-react, so they have common antigenic determinants. PMID- 2553737 TI - Adenylate cyclase toxin from Bordetella pertussis. Identification and purification of the holotoxin molecule. AB - Bordetella pertussis adenylate cyclase (AC) toxin is a calmodulin-activated adenylate cyclase enzyme which has the capacity to enter eukaryotic target cells and catalyze the conversion of endogenous ATP into cyclic AMP. In this work, the AC holotoxin molecule is identified and isolated. It is a single polypeptide of apparent 216 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Monoclonal antibodies which immunoprecipitate AC activity from extracts of wild type B. pertussis (BP338) react with this 216-kDa band on Western blots, and it is absent from a transposon Tn5 mutant (BP348) specifically lacking AC toxin. Isolation of the 216-kDa protein to greater than 85% purity by hydrophobic chromatography, preparative sucrose gradient centrifugation, and affinity chromatography using either calmodulin-Sepharose or monoclonal antibody coupled to Sepharose 4B yields stepwise increases in AC toxin potency, to a maximum of 88.3 mumol of cAMP/mg of target cell protein/mg of toxin. Electroelution of the 216-kDa band following sodium dodecyl sulfate polyacrylamide gel electrophoresis yields a preparation with both AC enzyme and toxin activities. These data indicate that this protein represents the AC holotoxin molecule. PMID- 2553738 TI - Intact DNA polymerase alpha/primase from mouse cells. Purification and structure. AB - A procedure is described for purification of DNA polymerase alpha/primase from cultured mouse lymphoblasts. Approximately 0.5 mg of enzyme, free of detectable contaminants, was obtained from 40 g of cells using seven conventional purification steps. The mouse enzyme contains subunits of 180, 70, 56, and 47 kDa, almost completely intact and similar to the subunit sizes reported for DNA polymerase alpha/primase from Drosophila embryos (Kaguni, L. S., Rossignol, J-M., Conaway, R. C., and Lehman, I. R. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 2221 2225) and Saccharomyces (Plevani, P., Foiani, M., Valsasnini, P., Badaracco, G., Cheriathundam, E., and Chang, L.M.S. (1985) J. Biol. Chem. 260, 7102-7107). A similar structure has also been inferred for mammalian DNA polymerase alpha/primase; however, previous descriptions of the highly purified mammalian enzyme complex have included substantial evidence of proteolysis and/or partial loss of subunits. In particular, the intact 180-kDa subunit has ordinarily been a minor component, and the molecular mass of the complex and total number of subunits have not been established. Results reported here indicate that the native DNA polymerase alpha/primase consists of one each of the four subunit sizes for a total of 353 kDa, based on estimates from denaturing gels, or 344 kDa when sizes deduced from available nucleic acid sequence data are substituted for three of the four subunits. A figure of 313 kDa was calculated from the sedimentation coefficient (8.9 S) and Stokes radius (81.1 A), the values for which also indicate a frictional ratio of 1.80, corresponding to an axial ratio of approximately 16 and suggesting a highly extended structure. PMID- 2553739 TI - Extrarenal nephroblastomas. AB - The cases of extrarenal nephroblastoma published in the world literature have been tabulated and reviewed, and are discussed from a historical point of view. PMID- 2553740 TI - Nucleus-specific and temporally restricted localization of proteins in Tetrahymena macronuclei and micronuclei. AB - Labeled nuclear proteins were microinjected into the cytoplasm of Tetrahymena thermophila. Macronuclear H1, calf thymus H1, and the SV40 large T antigen nuclear localization signal linked to BSA accumulated specifically in macronuclei, even if cells were in micronuclear S phase or were nonreplicating. The way in which histone H4 localized to either the macronucleus or the micronucleus suggested that it accumulates in whichever nucleus is replicating. The inability of the micronucleus to accumulate Tetrahymena H1 or heterologous nuclear proteins, even at a period in the cell cycle when it is accumulating H4, suggests that it has a specialized transport system. These studies demonstrate that although the mechanism for localizing proteins to nuclei is highly conserved among eukaryotes, it can differ between two porecontaining nuclei lying in the same cytoplasm. PMID- 2553741 TI - Transposition of domains between the M2 and HN viral membrane proteins results in polypeptides which can adopt more than one membrane orientation. AB - The influenza A virus M2 polypeptide is a small integral membrane protein that does not contain a cleaved signal sequence, but is unusual in that it assumes the membrane orientation of a class I integral membrane protein with an NH2-terminal ectodomain and a COOH-terminal cytoplasmic tail. To determine the domains of M2 involved in specifying membrane orientation, hybrid genes were constructed and expressed in which regions of the M2 protein were linked to portions of the paramyxovirus HN and SH proteins, two class II integral membrane proteins that adopt the opposite orientation in membranes from M2. A hybrid protein (MgMH) consisting of the M2 NH2-terminal and membrane-spanning domains linked precisely to the HN COOH-terminal ectodomain was found in cells in two forms: integrated into membranes in the M2 topology or completely translocated across the endoplasmic reticulum membrane and ultimately secreted from the cell. The finding of a soluble form suggested that in this hybrid protein the anchor function of the M2 signal/anchor domain can be overridden. A second hybrid which contained the M2 NH2 terminus linked to the HN signal anchor and ectodomain (MgHH) was found in both the M2 and the HN orientation, suggesting that the M2 NH2 terminus was capable of reversing the topology of a class II membrane protein. The exchange of the M2 signal/anchor domain with that of SH resulted in a hybrid protein which assumed only the M2 topology. Thus, all these data suggest that the NH2-terminal 24 residues to M2 are important for directing the unusual membrane topology of the M2 protein. These data are discussed in relationship to the loop model for insertion of proteins into membranes and the role of charged residues as a factor in determining orientation. PMID- 2553742 TI - Accessibility to proteases of the cytoplasmic G protein domain of vesicular stomatitis virus is increased during intracellular transport. AB - G1 and G2 are two forms of the membrane-integrated G protein of vesicular stomatitis virus that migrate differently in gel electrophoresis because G1 is modified by high-mannose and G2 by complex-type oligosaccharide side chains. The cytoplasmic domain in G1 is less exposed to cleavage by several proteases than in G2 molecules. Acylation by palmitic acid as well as inhibition of carbohydrate processing by swainsonine and deoxynojirimycin resulted in the same pattern of proteolytic sensitivity of both glycoproteins as in untreated cells. In contrast, accessibility of the cytoplasmic domain to proteases did not change when the intracellular transport of the G protein was blocked in carbonyl cyanide m chlorophenylhydrazone- or monensin-treated BHK-21 cells, respectively. The results suggest that the increase in accessibility of the cytoplasmic tail of the G protein occurs after the monensin block in the trans-Golgi and might reflect a conformational change of functional significance--i.e., making the cytoplasmic domain of the viral spike protein competent for its interaction with the viral core, inducing thereby the formation of the budding virus particle. PMID- 2553743 TI - Membrane domains of intestinal epithelial cells: distribution of Na+,K+-ATPase and the membrane skeleton in adult rat intestine during fetal development and after epithelial isolation. AB - The organization of the basolateral membrane domain of highly polarized intestinal absorptive cells was studied in adult rat intestinal mucosa, during development of polarity in fetal intestine, and in isolated epithelial sheets. Semi-thin frozen sections of these tissues were stained with a monoclonal antibody (mAb 4C4) directed against Na+,K+-ATPase, and with other reagents to visualize distributions of the membrane skeleton (fodrin), an epithelial cell adhesion molecule (uvomorulin), an apical membrane enzyme (aminopeptidase), and filamentous actin. In intact adult epithelium, Na+,K+-ATPase, membrane-associated fodrin, and uvomorulin were concentrated in the lateral, but not basal, subdomain. In the stratified epithelium of fetal intestine, both fodrin and uvomorulin were localized in areas of cell-cell contact at 16 and 17 d gestation, a stage when Na+,K+-ATPase was not yet expressed. These molecules were excluded from apical domains and from cell surfaces in contact with basal lamina. When Na+,K+-ATPase appeared at 18-19 d, it was codistributed with fodrin. Detachment of epithelial sheets from adult intestinal mucosa did not disrupt intercellular junctions or lateral cell contacts, but cytoplasmic blebs appeared at basal cell surfaces, and a diffuse pool of fodrin and actin accumulated in them. At the same time, Na+,K+-ATPase moved into the basal membrane subdomain, and extensive endocytosis of basolateral membrane, including Na+,K+-ATPase, occurred. Endocytosis of uvomorulin was not detected and no fodrin was associated with endocytic vesicles. Uvomorulin, along with some membrane-associated fodrin and some Na+,K+-ATPase, remained in the lateral membrane as long as intercellular contacts were maintained. Thus, in this polarized epithelium, interaction of lateral cell-cell adhesion molecules as well as basal cell-substrate interactions are required for maintaining the stability of the lateral membrane skeleton and the position of resident membrane proteins concentrated in the lateral membrane domain. PMID- 2553744 TI - Identification of actin nucleation activity and polymerization inhibitor in ameboid cells: their regulation by chemotactic stimulation. AB - Actin polymerization occurs in amebae of Dictyostelium discoideum after chemotactic stimulation (Hall, A. L., A. Schlein, and J. Condeelis. 1988. J. Cell. Biochem. 37:285-299). When cells are lysed with Triton X-100 during stimulation, an actin nucleation activity is detected in lysates by measuring the rate of pyrene-labeled actin polymerization. This stimulated nucleation activity is closely correlated with actin polymerization observed in vivo in its kinetics, developmental regulation, and cytochalasin D sensitivity. Actin polymerization is coordinate with pseudopod extension in synchronized populations of cells and is correlated with the accumulation of F actin in pseudopods. The stimulated actin nucleation activity is present in low-speed pellets from Triton lysates (cytoskeletons) within 3 s of stimulation and is stable compared with the nucleation activity of whole cell lysates. Low-speed supernatants contain a reversible inhibitor of the actin nucleation activity that is itself regulated by chemotactic stimulation. Neither activity requires Ca2+ and both are fully expressed in 10 mM EGTA. Fractions containing the inhibitor do not sever actin filaments but do inhibit actin polymerization that is seeded by fragments of purified F actin. These results indicate that chemotactic stimulation of Dictyostelium discoideum generates both an actin-nucleating activity and an actin polymerization inhibitor, and suggest that the parallel regulation of these two activities leads to the transient phases of actin polymerization observed in vivo. The different compartmentation of these two activities may account for polarized pseudopod extension in gradients of chemoattractant. PMID- 2553745 TI - Differential inhibition of nerve growth factor responses by purine analogues: correlation with inhibition of a nerve growth factor-activated protein kinase. AB - Purine analogues were used in this study to dissect specific steps in the mechanism of action of nerve growth factor (NGF). Protein kinase N (PKN) is an NGF-activated serine protein kinase that is active in the presence of Mn++. The activity of PKN was inhibited in vitro by purine analogues, the most effective of which was 6-thioguanine (apparent Ki = 6 microM). Several different criteria indicated that 6-thioguanine is not a general inhibitor of protein kinases and that it is relatively specific for PKN. For instance, it did not affect protein kinases A or C and was without effect on the overall level and pattern of protein phosphorylation by either intact or broken PC12 cells. Since purine analogues rapidly and effectively enter cells, they were also assessed for their actions on both transcription-dependent and -independent responses of PC12 cells to NGF. NGF promoted neurite regeneration was reversibly suppressed by the analogues and at concentrations very similar to those that inhibit PKN. Comparable concentrations of the analogues also blocked NGF-stimulated induction of ornithine decarboxylase activity. In contrast to its inhibition of neurite regeneration and ornithine decarboxylase induction, 6-thioguanine did not suppress NGF-dependent induction of c-fos mRNA expression. Thus, purine analogues such as 6-thioguanine appear capable of differentially suppressing some, but not other actions of NGF. These findings suggest the presence of multiple pathways in the NGF mechanism and that these can be dissected with purine analogues. Moreover, these data are compatible with a role for protein kinase N in certain of these pathways. PMID- 2553746 TI - In vivo analysis of glial cell phenotypes during a viral demyelinating disease in mice. AB - C57 BL/6N mice injected intracranially with the A59 strain of mouse hepatitis virus exhibit extensive viral replication in glial cells of the spinal cord and develop demyelinating lesions followed by virus clearing and remyelination. To study how different glial cell types are affected by the disease process, we combine three-color immunofluorescence labeling with tritiated thymidine autoradiography on 1-micron frozen sections of spinal cord. We use three different glial cell specific antibodies (a) to 2',3' cyclic-nucleotide 3' phosphohydrolase (CNP) expressed by oligodendrocytes, (b) to glial fibrillary acidic protein (GFAP) expressed by astrocytes, and (c) the O4 antibody which binds to O-2A progenitor cells in the rat. These progenitor cells, which give rise to oligodendrocytes and type 2 astrocytes and react with the O4 antibody in the adult central nervous system, were present but rare in the spinal cord of uninfected mice. In contrast, cells with the O-2A progenitor phenotype (O4 + only) were increased in number at one week post viral inoculation (1 WPI) and were the only immunostained cells labeled at that time by a 2-h in vivo pulse of tritiated thymidine. Both GFAP+ only and GFAP+, O4+ astrocytes were also increased in the spinal cord at 1 WPI. Between two and four WPI, the infected spinal cord was characterized by the loss of (CNP+, O4+) oligodendrocytes within demyelinating lesions and the presence of O-2A progenitor cells and O4+, GFAP+ astrocytes, both of which could be labeled with thymidine. As remyelination proceeded, CNP immunostaining returned to near normal and tritiated thymidine injected previously during the demyelinating phase now appeared in CNP+ oligodendrocytes. Thus O4 positive O-2A progenitor cells proliferate early in the course of the demyelinating disease, while CNP positive oligodendrocytes do not. The timing of events suggests that the O-2A progenitors may give rise to new oligodendrocytes and to type 2 astrocytes, both of which are likely to be instrumental in the remyelination process. PMID- 2553747 TI - States of developmental commitment of a mouse embryonal carcinoma cell line differentiating along a neural pathway. AB - The embryonal carcinoma cell line PCC7-S-AzaR1 (clone 1009) has been shown to differentiate in the presence of all-trans retinoic acid and dibutyryl cAMP into cells of predominantly neural properties (Paulin, D., H. Jakob, F. Jacob, K. Weber, and M. Osborn. 1982. Differentiation. 22:90-99). By analyzing the marker expression of derivatives in further detail, we characterized the two major cell phenotypes as neuron- and fibroblast-like and the two minor ones as astroglia- and endothelial-like. The stability of developmental commitment of clone 1009 was tested by recloning. The isolated subclones exhibited different patterns of chemically induced derivatives, with some of them (denoted N-clones) producing only a single (neuronal) cell type. As shown by long-term cultures in the absence of retinoic acid, the properties of isolated subclones remained essentially stable. In contrast to the clones producing neuron-like and other derivatives upon induced differentiation, the (exclusively neuronal) derivatives of N-clones detached and died within a few days in culture. If maintained in the presence of other neural cell types, however, their survival was dramatically extended indicating a requirement for specific interactions with other cells of the same tissue. The patterns of derivatives obtained from N-clones depended on the chemical nature of the substrate on which they were grown. Thus, when seeded on laminin-coated surfaces before induced differentiation, N-clones developed not only to neuron-like derivatives but rather to the same four derivatives observed with the original cell pool. These and further results suggest a common cell lineage of the identified phenotypes. The isolated subclones of uninduced cells probably represent different states of commitment within the same developmental pathway. Their stability offers the opportunity to analyze the nature of cellular commitment on the cellular, molecular, and genetic levels. This makes the family of clones derived from PCC7-S-AzaR1 (clone 1009) cells an advantageous in vitro model of mammalian brain early ontogenesis. PMID- 2553748 TI - Signal transduction by epidermal growth factor occurs through the subclass of high affinity receptors. AB - Many cell types display two classes of epidermal growth factor receptor (EGFR) as judged from EGF binding studies; i.e., a major class of low affinity EGFR and a minor class of high affinity EGFR. We have studied their respective contribution to the cascade of events elicited by EGF in human A431 carcinoma cells, using anti-EGFR mAb 2E9. This antibody specifically blocks EGF binding to low affinity EGFR, without activating receptors in intact cells, and thus enables us to study the effects of exclusive EGF binding to high affinity EGFR. We show that blocking of low affinity EGFR by mAb 2E9 has almost no effect on the activation of the receptor protein-tyrosine kinase by EGF, suggesting that EGFR kinase activation occurs exclusively through the subclass of high affinity EGFR (5-10%). In addition, we provide evidence that high affinity EGFR exists both in monomeric and dimeric forms, and that cross-phosphorylation of low affinity EGFR by high affinity EGFR may take place in dimers of both receptor types. We demonstrate that the following early cellular response to EGF are also unimpaired in the presence of mAb 2E9: (a) inositol phosphate production, (b) release of Ca2+ from intracellular stores, (c) rise in intracellular pH, (d) phosphorylation of EGF on threonine residue 654, (e) induction of c-fos gene expression, and (f) alteration in cell morphology. As possible nonspecific side effects, we observed that the EGF induced Ca2+ influx and fluid-phase pinocytosis were inhibited in A431 cells in the presence of mAb 2E9. We conclude, therefore, that the activation of the EGFR signal transduction cascade can occur completely through exclusive binding of EGF to the subclass of high affinity EGFR. PMID- 2553750 TI - Expression of Na,K-ATPase alpha subunit isoforms in the human ciliary body and cultured ciliary epithelial cells. AB - We have analyzed the expression of Na,K-ATPase alpha subunit isoforms in the transporting ciliary processes of the human eye and in cultured cells derived from non-pigmented (NPE) and pigmented (PE) ciliary epithelium. Northern hybridization analysis shows that the mRNAs encoding all the three distinct forms of Na,K-ATPase alpha subunit [alpha 1, alpha 2, and alpha 3] are expressed in the human ciliary processes in vivo. Immunohistochemical analysis using antibodies specific for each of the three alpha subunit isoforms confirms that these polypeptides are present in the microsomal fraction from the human ciliary processes. The monoclonal antibody McB2, which is specific to the Na,K-ATPase alpha 2 subunit isoform, has been found to decorate specifically the basolateral membrane domains of NPE cells but not of the PE cells, suggesting its expression in vivo only in the ocular NPE ciliary epithelium. However, cultured cells derived from the NPE and PE layers exhibit a different pattern of expression of mRNA and protein for the Na,K-ATPase alpha subunit isoforms when compared to the tissue. Both the NPE and PE cells express alpha 1 and alpha 3 mRNA and polypeptide, whereas alpha 2 mRNA and polypeptide are undetectable in these cells. The established cell lines derived from the NPE layer express comparable levels of the alpha 1 and alpha 3 isoforms of Na,K-ATPase as detected in the primary culture. However, the established NPE cell lines are also distinguishable from the normal PE cells when analyzed by Western blot analysis with A x 2 antibodies. The results presented here clearly show that the NPE and PE cells in the ciliary body have a distinct expression of Na,K-ATPase alpha subunit isoforms as compared to cultured cells. PMID- 2553749 TI - Transformation of NIH 3T3 cells with basic fibroblast growth factor or the hst/K fgf oncogene causes downregulation of the fibroblast growth factor receptor: reversal of morphological transformation and restoration of receptor number by suramin. AB - When NIH 3T3 cells were transfected with the cDNA for basic fibroblast growth factor (bFGF), most cells displayed a transformed phenotype. Acquisition of a transformed phenotype was correlated with the expression of high levels of bFGF (Quarto et al., 1989). Cells that had been transformed as a result of transfection with bFGF cDNA had a decreased capacity to bind 125I-bFGF to high affinity receptors. NIH 3T3 cells transfected with bFGF cDNA that expressed lower levels of bFGF were not transformed and had a normal number of bFGF receptors. NIH 3T3 cells transfected with the hst/Kfgf oncogene, which encodes a secreted molecule with 45% homology to bFGF, also displayed a transformed phenotype and decreased numbers of bFGF receptors. However, NIH 3T3 cells transfected with the H-ras oncogene were transformed but had a normal number of bFGF receptors. Thus, transformation by bFGF-like molecules resulted in downregulation of bFGF receptors. Receptor number was not affected by cell density for both parental NIH 3T3 cells and transformed cells. In the cells transfected with bFGF cDNA that were not transformed, the receptors could be downregulated in response to exogenous bFGF. Conditioned medium from transformed transfected cells contained sufficient quantities of bFGF to downregulate bFGF receptors on parental NIH 3T3 cells. Thus, the downregulation of bFGF receptors seemed related to the presence of bFGF in an extracytoplasmic compartment. Treatment of the transformed transfected NIH 3T3 cells with suramin, which blocks the interaction of bFGF with its receptor, reversed the morphological transformation and restored receptors almost to normal numbers. These results demonstrate that in these cells bFGF transforms cells by interacting with its receptor and that bFGF and hst/K-fgf may use the same receptor. PMID- 2553751 TI - Evidence for increased alpha MSH receptor binding in the F1 variant of B16 melanoma cells grown in dialyzed fetal calf serum. AB - The specific binding of an alpha MSH analogue (Ac-[Nle4, D-Phe7] alpha MSH4-11 NH2) was enhanced in the presence of 10% dialyzed fetal calf serum (FCS) as compared with 10% FCS (nondialyzed) in the F1 variant of B16 melanoma cells. The replenishment of dialyzed serum with adrenocorticotrophic hormone (ACTH) or insulin had no effect on the increased level of alpha MSH receptor binding in these cells. However, 10 nM alpha MSH or 1 microM ACTH under identical conditions significantly decreased the level of alpha MSH binding. Competitive binding studies involving the alpha MSH analogue revealed that the specificity of the receptor was restricted to the complete molecule of alpha MSH, our analogue, beta MSH and ACTH1-24, ACTH4-10, which contains the amino acid sequence responsible for biological activity, showed a very low affinity for the receptor. Furthermore, we observed an interesting phenomenon unique to dialyzed FCS in that once the cells were grown to confluence and melanin was produced, the cells were no longer viable. However, in McCoy's medium, which is deficient in tyrosine, the cells did not produce melanin and remained viable. PMID- 2553752 TI - Activation of the Na+/H+ exchanger by phorbol ester and osmotic shock is dependent on the degree of neutrophilic maturation. AB - Activation of neutrophils leading to superoxide production is accompanied by cytoplasmic alkalinization, which results from stimulation of the Na+/H+ exchanger. Since the exchanger undergoes permanent alterations during neutrophilic maturation of HL-60 cells (Costa-Casnellie et al.: Journal of Biological Chemistry 263:11851-11855, 1988), we investigated whether its response to external stimuli such as phorbol esters or osmotic shock also was modified during cell maturation. Mature HL-60 cells produce superoxide in response to active phorbol esters, whereas immature HL-60 cells do not. Stimulation of the exchanger by active phorbol esters (phorbol 12-myristate 13-acetate or phorbol 12,13-dibutyrate) was observed in mature neutrophilic HL-60 cells but not in their immature counterparts. Inactive 4-alpha phorbol had no effect in either cell population. Compound H7 inhibited phorbol ester activation by 65%. In mature neutrophilic cells activation of the exchanger by phorbol esters caused two novel changes of its properties: 1) its apparent Km for Na+ transport increased 2-fold; 2) its Vmax increased 2.6-fold. Phorbol esters also caused a shift in pH dependence of activation similar to that induced in other cells. Osmotic shock, a different method known to activate the exchanger of other cells, induced activation in mature neutrophilic cells but not in immature cells. Thus, the response of the exchanger to external stimuli is affected by alterations occurring in association with cell maturation. PMID- 2553753 TI - Heat shock stimulates the release of arachidonic acid and the synthesis of prostaglandins and leukotriene B4 in mammalian cells. AB - Heat shock has a profound influence on the metabolism and behavior of eukaryotic cells. We have examined the effects of heat shock on the release from cells of arachidonic acid and its bioactive eicosanoid metabolites, the prostaglandins and leukotrienes. Heat shock (42-45 degrees) increased the rate of arachidonic acid release from human, rat, murine, and hamster cells. Arachidonate accumulation appeared to be due, at least partially, to stimulation of a phospholipase A2 activity by heat shock and was accompanied by the accumulation of lysophosphatidyl-inositol and lysophosphatidylcholine in membranes. Induction of arachidonate release by heat did not appear to be mediated by an increase in cell Ca++. Stimulation of arachidonate release by heat shock in hamster fibroblasts was quantitatively similar to the receptor-mediated effects of alpha thrombin and bradykinin. The effects of heat shock and alpha thrombin on arachidonate release were inhibited by glucocorticoids. Increased arachidonate release in heat-shocked cells was accompanied by the accelerated accumulation of cyclooxygenase products prostaglandin E2 and prostaglandin F2 alpha and by 5-lipoxygenase metabolite leukotriene B4. Elevated concentrations of arachidonic acid and metabolites may be involved in the cytotoxic effects of hyperthermia, in homeostatic responses to heat shock, and in vascular and inflammatory reactions to stress. PMID- 2553755 TI - Viral infections. AB - This article has concentrated on the etiology, pathogenesis, diagnosis, and treatment of viral infections of the hand. Some of these entities are quite common, but are often misdiagnosed. Herpes simplex appears to be the most common viral infection involving the hand. These and other viral infections can have much graver consequences in immunosuppressed patients. Recurrent episodes are common in herpetic and other viral infections. Health care workers have long been at risk, but improved compliance with prophylactic measures would seem to be decreasing the number of cases in this population. A plea has been made to disregard the term herpetic whitlow because it is an inaccurate description of the lesion and implies that an inappropriate type of treatment is required. Bacterial whitlows or felons require incision and drainage of the deep pulp space. Herpetic infections in this area do not. Less common infections such as cowpox, pseudocowpox (milkers nodules), ORF, and coxsackievirus (HFMD) infection of the hand have been brought to the attention of the reader. The bothersome warts caused by the human papillomavirus have been described and the systemic ramifications of hand-to-hand contact and hand injury causing more serious viral problems has been mentioned. A common thread in the care of the patient with these types of diseases is that an adequate history and physical remain invaluable in arriving at the correct diagnosis. With this correct diagnosis all of us, as physicians, then can easily abide by one of our basic principles Primum non nocere, "Let me help--but first let me do no harm." PMID- 2553754 TI - Cyclic adenosine monophosphate acts synergistically with dexamethasone to inhibit the entrance of cultured adult rat hepatocytes into S-phase: with a note on the use of nucleolar and extranucleolar [3H]-thymidine labelling patterns to determine rapid changes in the rate of onset of DNA replication. AB - Analogs of cyclic adenosine monophosphate (cAMP) (N6benzoyl cAMP and N6monobutyryl cAMP) as well as agents that increased the intracellular level of cAMP (glucagon and isobutylmethylxanthine) inhibited the EGF-stimulated DNA replication of adult rat hepatocytes in primary culture independently of cell density. This inhibition was strongly potentiated by the glucocorticoid dexamethasone. The effect of cAMP (and dexamethasone) was not due to toxicity, because the inhibition was reversible and the cell ultrastructure preserved. cAMP acted by decreasing the rate of transition from G1- to S-phase, the duration of G2- and S-phase of the hepatocyte cell cycle being unaffected. DNA replication started in the extranucleolar compartment of the nucleus and ended in the nucleolar compartment as described earlier for cells grown in the absence of cAMP (O.K. Vintermyr and S.O. Doskeland, J. Cell. Physiol., 1987, 132:12-21). The action of cAMP was very rapid: significant inhibition of the transition was noted 2 hr after the addition of glucagon/IBMX and half-maximal inhibition after 4 hours. The determination of extranucleolarly labelled nuclei in cells pulse labelled with [3H]thymidine allowed precise analysis of rapid changes in the probability of transition from G1- to S-phase. The extranucleolar labelling index could also be determined in cells continuously exposed to [3H]thymidine. PMID- 2553756 TI - [Value of the peroperative association of echography, extemporaneous cytology and immediately available insulin blood levels of the portal vein. Apropos of 2 cases of insulinoma]. AB - The authors report two cases of insulinoma managed by per-operative ultrasound and cytology, together with immediate portal vein insulin levels. They stress the value of these per-operative exploratory techniques which reduce the number of pre-operative invasive examinations while ensuring a greater efficacy in the localization and treatment of these tumors. PMID- 2553757 TI - Sun exposure for the elderly. PMID- 2553758 TI - Inherited susceptibility to colorectal cancer. AB - Colorectal cancer and its potential precursor, adenomatous polyps, tend to aggregate in families, but it is not clear why. Heritability is established for only rare colorectal cancers, some prominently involving polyposis. Preliminary studies suggest, however, that susceptibility to the common colorectal cancer precursors, "sporadic" adenomatous polyps, is also inherited. PMID- 2553759 TI - Optimization of size-exclusion separation of proteins on a Superose column. PMID- 2553760 TI - Analysis of a novel cardiac stimulant, UK-61 260, in human plasma by gas chromatography-mass spectrometry with selected-ion monitoring. PMID- 2553761 TI - Simultaneous determination of propoxyphene and norpropoxyphene in biological samples by gas chromatography using an on-column injection technique with a fused silica capillary column. PMID- 2553762 TI - High-performance liquid chromatographic assay of k-opioid selective benzeneacetamide derivatives (U50488, U69593 and PD117302) in rat plasma and brain. PMID- 2553764 TI - Synthesis of prolyl 4-hydroxylase by cultured chicken corneal cells. AB - Prolyl hydroxylase (EC 1.14.11.2) synthesis was examined in the cultured corneas of 17-day-old chick embryos which were cultured in F-12 medium supplemented with fetal calf serum. The amount of prolyl hydroxylase in cells was measured by immunoelectrophoresis using antibodies directed against this enzyme. The results indicated that the amounts of prolyl hydroxylase depended on cell density. The cells were harvested at the early-log phase and plated at high cell density to mimic the culture condition of cells at the stationary phase (crowding). The amount of 35S-prolyl hydroxylase indicated a two-fold increase in the rate of prolyl hydroxylase synthesis after crowding. By use of 32P-labeled cDNA for the beta-subunit of prolyl hydroxylase a proportionate increase of the mRNA was noted in accordance with the increase in the rate of prolyl hydroxylase synthesis, suggesting that this enzyme synthesis is regulated at the transcriptional level. PMID- 2553763 TI - Application of inclusion chromatography to the determination of in vitro metabolites of estriol. AB - The application of inclusion chromatography with beta-cyclodextrin as a mobile phase additive in high-performance liquid chromatography to the determination of in vitro metabolites of estriol is reported. Compared to conventional methods, the inclusion chromatography gives much more satisfactory separation of isomeric estriol derivatives in a short time. Species difference (rat and guinea pig liver homogenates) is observed in the glucuronidation of estriol. The hydroxylation of estriol with rat liver homogenate occurs preferentially at the 2-position. Methylation of this product with catechol-O-methyl transferase in rat liver gives almost equal amounts of 2- and 3-methyl ethers. In contrast, 4-hydroxyestriol gives 4-methyl ether as a main product. PMID- 2553765 TI - Rotavirus infection of Thai infants. Serotypes and subgroups of cultivated viruses in 1984 and 1985. PMID- 2553766 TI - Direct detection of cytomegalovirus from bronchoalveolar lavage samples by using a rapid in situ DNA hybridization assay. AB - An in situ DNA hybridization assay was compared with centrifugation culture for rapid detection of cytomegalovirus (CMV) from bronchoalveolar lavage (BAL) samples. Eighty BAL samples were inoculated into both centrifugation culture and standard culture. Cytospin preparations of the BAL samples were studied in a 75 min in situ DNA hybridization assay using the PathoGene CMV kit (Enzo Biochem, Inc., New York, N.Y.). Of the 80 samples, 39 (49%) were positive for CMV; 37 of 39 (95%) were positive by centrifugation culture, 34 of 39 (87%) were positive in standard culture, 24 of 39 (62%) were positive by in situ hybridization, and 20 of 39 (56%) were positive by histologic and/or immunofluorescence techniques. The in situ hybridization assay detected 23 of the 37 samples positive in centrifugation culture, for a sensitivity of 62% and a specificity of 98%. We conclude that the in situ hybridization assay is a specific and more rapid test than centrifugation culture and standard culture for diagnosis of CMV pulmonary infection. For the clinical laboratory, however, current hybridization methods are not sufficiently sensitive to replace centrifugation culture for detection of CMV in BAL specimens. PMID- 2553767 TI - Detection of infectious bursal disease viruses by using cloned cDNA probes. AB - A molecular clone representing 445 base pairs at the 3' end of infectious bursal disease virus (IBDV) genome segment B was used in a dot blot hybridization assay to detect viral RNA from cell culture and from chicken bursa and spleen tissue specimens. The cloned nucleotide sequence represents approximately 14% of the virus-encoded polymerase (VP-1) gene. The lower detection limit of radiolabeled probes prepared from this clone was 0.1 ng of IBDV double-stranded RNA. The probe had broad specificity and was used to detect four serotype 1 IBDV strains and one serotype 2 IBDV strain. This probe, however, did not cross-react with nucleic acid extracted from nine unrelated poultry viruses. A rapid procedure for isolation of IBDV genomic RNA from bursa and spleen tissue specimens was developed and used with the dot blot hybridization assay to detect IBDV strains in tissue samples from experimentally infected and commercially reared chickens. PMID- 2553768 TI - Differentiation of primary from nonprimary genital herpesvirus infections by detection of polymeric immunoglobulin A activity. AB - We analyzed antibody responses to herpes simplex virus (HSV) in 18 patients by using sucrose gradient centrifugation and an enzyme-linked immunosorbent assay for antibody determination. We found that the HSV-specific immunoglobulin A (IgA) antibody in early-convalescent-phase serum specimens from all of 8 primary cases consisted solely of polymeric forms, whereas in all 10 nonprimary cases, the major component was monomeric. These results indicate that the ratio of polymeric IgA to monomeric IgA antibody activity may be useful for differentiating primary from nonprimary HSV infections. PMID- 2553769 TI - Serological characterization of bovine rotaviruses isolated from dairy and beef herds in Argentina. AB - Bovine rotaviruses isolated from beef and dairy herds in Argentina were serotyped by the immunoperoxidase focus reduction assay as previously described (G. Gerna, M. Battaglia, G. Milenesi, N. Passarani, E. Percivalle, and E. Cattaneo, Infect. Immun. 43:722-729, 1984). Three strains from beef herds were related to the UK and NCDV bovine rotavirus strains defined as serotype 6 (Y. Hoshino, R. G. Wyatt, H. B. Greenberg, J. Flores, and A. Z. Kapikian, J. Infect. Dis. 149:694-702, 1984). Two other strains from dairy herds were classified as bovine viruses related to the bovine B223 strain reported by Woode and co-workers (G. N. Woode, N. E. Kelso, T. F. Simpson, S. K. Gaul, L. E. Evans, and L. Babiuk, J. Clin. Microbiol. 18:358-364, 1983) in the United States. A serotyping antibody-capture enzyme-linked immunoassay to detect serotype 6 rotavirus using a serotype 6 specific monoclonal antibody was developed and evaluated for strain characterization. Characterization of 72 group A rotavirus-positive fecal samples from beef herds and 43 fecal samples from dairy herds showed a predominance of serotype 6 rotavirus in beef herds but both serotype 6 and non-serotype 6 rotaviruses in dairy herds. Analysis of genomic double-stranded RNA by polyacrylamide gel electrophoresis showed that when outbreaks were caused by one serotype only a single electropherotype was present in all samples. PMID- 2553770 TI - Beta-endorphin alters a viral induced central nervous system disease in normal mice but not in nude mice. AB - A single intracerebroventricular injection of 100 ng of beta-endorphin altered the course of the central nervous system (CNS) infection of a temperature sensitive mutant of vesicular stomatitis virus (VSV), tsG31-KS5. When mice were administered beta-endorphin and then 24 h later infected intracerebrally with tsG31-KS5 VSV, 70% of the animals died within 8 days of infection. In comparison, less than 10% of the animals had died after 21 days when infected with tsG31-KS5 VSV alone. When mice were injected with beta-endorphin and tsG31-KS5 VSV simultaneously, or with beta-endorphin 21 days after infection, the more aggressive clinical disease was not observed. Superficially, the more lethal disease induced by beta-endorphin appeared to be a result of a mild hypothermia caused by the neuropeptide. beta-Endorphin, however, did not influence the disease in nude (nu/nu) mice even though their core temperatures were reduced to an extent similar to that of BALB/c (+/+) mice, implicating the involvement of T lymphocytes in the alteration of the course of infection in normal mice. PMID- 2553771 TI - Characterization of the local and systemic B cell response of normal and athymic nude mice with Sindbis virus encephalitis. AB - During Sindbis virus (SV) encephalitis in mice B cells are an important component of the mononuclear inflammatory response and recovery depends primarily on the development of antiviral antibody. To begin to characterize various parameters of the local B cell response during SV encephalitis we have defined B cell isotype expression in brain sections, splenocytes and peripheral blood mononuclear cells (PBMC) in normal and athymic nude mice using an immunoperoxidase technique. Early (days 3-5) in SV encephalitis brain perivascular B cells are IgM or IgM/IgD bearing lymphocytes, later (days 10-14) most B cells express one of the IgG isotypes or IgA. The pattern of isotype expression seen in the brain during the course of the encephalitis is reflected in the spleen and blood. The data suggest that progressive isotype switching results in an increasingly higher percentage of certain isotypes, especially IgG2a. Isotype switching of most B cells may occur outside of the brain, or may arise in situ from the IgM/IgD-bearing B cells found in the brain throughout the course of encephalitis. In athymic nude mice numbers of B cells in brain were markedly decreased and the cells present were primarily IgM-bearing, although IgG isotypes and IgA did appear late (day 14). The data suggest that T cells are required for recruitment of B cells into the inflammatory response as well as for normal isotype switching and peripheral B cell maturation during SV encephalitis. PMID- 2553772 TI - In vitro synthesis of antibodies to acetylcholine receptor by Epstein-Barr virus stimulated B-lymphocytes derived from patients with myasthenia gravis. AB - Peripheral blood lymphocytes and B-cells were obtained from patients with myasthenia gravis and stimulated in vitro with either pokeweed mitogen or Epstein Barr virus (EBV), respectively. EBV stimulation of B-cells caused a production of antibodies to acetylcholine receptor in 15 of the 25 myasthenia gravis patients: the EBV stimulation of B-cells was more effective in this regard than the pokeweed mitogen stimulation of peripheral blood lymphocytes. The in vitro synthesis of anti-acetylcholine receptor antibodies was found to be positively correlated with both the patients' sera antibody titers and with the disease severity. PMID- 2553773 TI - Human neutrophil gelatinase is a component of specific granules. AB - Previous investigators have proposed that gelatinase, a metalloproteinase found in neutrophils, is stored in a novel secretory compartment distinct from the two major granule populations, azurophilic and specific. To locate this proteinase in human neutrophils we reacted the cells for peroxidase and then applied monospecific polyclonal antibodies to human neutrophil gelatinase to immunolabel ultrathin frozen sections using an immunogold technique. Gelatinase was localized in a population of peroxidase-negative granules. Double-labeling experiments using antibodies against lactoferrin, a marker for specific granules, and gelatinase demonstrated colocalization of the two antigens in 80% of the specific granules. However, some granules immunostained with only the lactoferrin or gelatinase antibody. Similar techniques were used to examine precursor cells from bone marrow. In myelocytes both gelatinase and lactoferrin were present in large developing specific granules; however, some mature specific granules contained only lactoferrin. Thus, it is possible that lactoferrin synthesis begins earlier than gelatinase synthesis and that overlapping synthesis and segregation occurs during the myelocyte stage. These findings suggest that the main storage compartment of gelatinase is within the peroxidase-negative specific granules. PMID- 2553774 TI - Blockade of the type I somatomedin receptor inhibits growth of human breast cancer cells in athymic mice. AB - Insulin and insulin-like growth factors (IGIs) stimulate the growth of human breast cancer cells in vitro. The type I somatomedin receptor (SR) expressed in these cells may mediate the growth effects of these peptides. We have examined the role of this receptor on human breast cancer growth with a monoclonal antibody (alpha-IR-3) that blocks the receptor binding domain and inhibits IGF-I induced growth. alpha-IR-3 inhibited clonal growth in vitro and blocked the mitogenic effect of exogenous IGF-I in both MCF-7 and MDA-231 breast cancer cell lines. Antibody-induced blockade of the type I SR also inhibited the estrogen independent MDA-231 cells growing in vivo in nude mice, but growth of the estrogen-dependent MCF-7 cells was unaffected. IGIs are important growth regulators of MDA-231 breast cancer cells. Blockade of this growth stimulatory pathway may provide a new treatment strategy. PMID- 2553775 TI - Phagocytosis of aged human neutrophils by macrophages is mediated by a novel "charge-sensitive" recognition mechanism. AB - The removal of neutrophils and their histotoxic contents from the inflamed site is a prerequisite for resolution of tissue injury, and a point at which factors critical to the pathogenesis of chronic inflammation may act. Engulfment of intact, senescent neutrophils by macrophages represents an important neutrophil disposal process. In this study the mechanism by which human monocyte-derived macrophages (M phi) recognized and ingested human neutrophils that had been aged in culture was studied using an in vitro phagocytic assay. Inhibition of M phi receptors for Ig Fc and the opsonic complement fragments C3b and iC3b with MAbs to M phi FcR, CR1, CR3, and CR4 had no effect on recognition, and the pattern of inhibition observed when polyanions were included in the medium at 1 mg/ml was different from that reported for the M phi receptor for protein advanced glycosylation end products (AGE), indicating a recognition mechanism different from those proposed for M phi phagocytosis of senescent erythrocytes. Furthermore, although aging neutrophils undergo programmed cell death (or apoptosis), which is directly related to recognition by M phi, the pattern of inhibition observed with monosaccharides was different from that reported to inhibit the binding of apoptotic mouse thymocytes to isologous M phi. By contrast, evidence was obtained for a novel recognition mechanism inhibitable by cationic sugars and amino acids in a charge-dependent fashion, and directly modulated by pH but not affected by inhibitors of the mannose-6-phosphate, sheep erythrocyte, mannosyl-fucosyl, asialoglycoprotein, and scavenger receptors of the macrophage. These observations suggest that hydrogen ions and charged molecules may modulate M phi uptake of senescent neutrophils at inflamed sites, and that recognition itself may involve charged structures on the cells. PMID- 2553776 TI - Synthesis of stress proteins is increased in individuals with homozygous PiZZ alpha 1-antitrypsin deficiency and liver disease. AB - Individuals who are homozygous for the protease inhibitor phenotype Z (PiZ) genetic variant of alpha 1-antitrypsin (alpha 1-AT) have reduced plasma concentrations of alpha 1-AT, and are susceptible to premature development of pulmonary emphysema. A subset of this population develops chronic liver disease. The reduction in plasma concentrations of alpha 1-AT results from a selective defect in secretion as the abnormal PiZ alpha 1-AT protein accumulates within the cell. It has recently been shown in several experimental systems that the heat shock/stress response, a response characterized by the synthesis of a family of highly evolutionarily conserved proteins during thermal or chemical stress, may also be activated by the presence of abnormal proteins within the cell. Therefore, we predicted that the heat shock/stress response would be induced in the absence of thermal or chemical stress in alpha 1-AT-synthesizing cells of PiZZ individuals. In the following study, however, we show that net synthesis of proteins in the heat shock/stress gene family (SP90, SP70, ubiquitin) is increased only in a subset of the population, PiZZ individuals with liver disease. It is not significantly increased in PiZZ individuals with emphysema or in those without apparent tissue injury. Net synthesis of stress proteins is not increased in individuals with another variant of the alpha 1-AT gene (PiS alpha 1 AT) and is not increased in individuals with severe liver disease but a normal alpha 1-AT haplotype (PiM alpha 1-AT). These results demonstrate that the synthesis of stress proteins is increased in a subset of individuals with homozygous PiZZ alpha 1-AT deficiency, those also having liver disease. PMID- 2553777 TI - Effects of leukotriene B4 in the human lung. Recruitment of neutrophils into the alveolar spaces without a change in protein permeability. AB - Leukotriene B4 (LTB4) is a major product of human alveolar macrophages and has potent chemotactic activity for neutrophils (PMN) in vitro. To evaluate the effects of LTB4 in the normal human lung, we instilled LTB4 (5 X 10(-7)M, 10 ml) into a subsegment of the right middle lobe and 0.9% NaCl (10 ml) into a subsegment of the lingula using a fiberoptic bronchoscope in 12 healthy human volunteers. 4 h later, we performed bronchoalveolar lavage of the same subsegments. Compared with the NaCl instillation, LTB4 caused a large increase in lavage total cells (NaCl = 6.8 +/- 1.0 X 10(6) vs. LTB4 = 26.4 +/- 5.0 X 10(6), P less than 0.01), most of which were PMN (NaCl = 12.2 +/- 4.6% vs. LTB4 = 55.7 +/- 6.0%, P less than 0.001). In contrast, there was only a small increase in lavage total protein, and the lavage total protein correlated weakly with lavage total cells and PMN. The production of superoxide anion by the lavage PMN in response to phorbol myristate acetate was similar to that of peripheral blood PMN. The migration of lavage PMN was normal toward the chemotactic peptide FMLP, but reduced toward LTB4 and zymosan-activated human serum. Morphometric analysis using transmission electron microscopy indicated a selective loss of small granules in the lung neutrophils as compared with peripheral blood neutrophils. The data indicate that in the normal human lung, LTB4 can recruit active PMN into the airspaces without causing a significant change in the protein permeability of the epithelial barrier. PMID- 2553778 TI - Marked QRS complex abnormalities and sodium channel blockade by propoxyphene reversed with lidocaine. AB - The opiate analgesic propoxyphene produces cardiac toxicity when taken in overdose. We recently observed a patient with propoxyphene overdose in whom marked QRS widening was reversed by lidocaine. The reversal is apparently paradoxical as both agents block the inward sodium current (INa). We examined possible mechanisms of the reversal by measuring INa in rabbit atrial myocytes during exposure to propoxyphene and the combination of propoxyphene and lidocaine (60 and 80 microM, respectively). Propoxyphene caused use-dependent block of INa during pulse train stimulation. Block recovered slowly with time constants of 20.8 +/- 3.9 s. Block during lidocaine exposure recovered with time constants of 2-3 s. During exposure to the mixture, block recovered as a double exponential. The half time for recovery during exposure to the mixture was 1.6 +/- .9 s compared with a half-time of 14.3 +/- 2.9 s during exposure to propoxyphene alone. During pulse train stimulation, less steady-state block was observed during exposure to the mixture than during exposure to propoxyphene alone when the interval between pulses was greater than 0.95 s. Both drugs compete for a common receptor during the polarizing phase. The more rapid dissociation of lidocaine during the recovery period leads to less block during the mixture than during exposure to propoxyphene alone. The experiments suggest a mechanism for reversal of the cardiac toxicity of drugs which have slow unbinding kinetics. PMID- 2553779 TI - Damage to the bases in DNA induced by stimulated human neutrophils. AB - Leukocyte-induced DNA damage may partially account for the known association between chronic inflammation and malignancy. Since elucidation of the chemical nature of leukocyte-induced DNA damage may enhance our understanding of the mechanisms underlying leukocyte-induced DNA damage and the carcinogenesis associated with inflammation, the present study was undertaken to characterize the chemical modifications that occur in DNA exposed to stimulated human neutrophils. Calf thymus DNA was exposed to phorbol myristate acetate (PMA) stimulated neutrophils in the presence or absence of exogenously added iron ions. DNA samples were subsequently hydrolyzed, derivatized and analyzed by gas chromatography-mass spectrometry with selected-ion monitoring. A variety of base modifications including cytosine glycol, thymine glycol, 4,6-diamino-5 formamidopyrimidine, 8-hydroxyadenine, 2,6-diamino-4-hydroxy-5 formamidopyrimidine, and 8-hydroxyguanine were identified. The yield of these various base products was increased by the addition of iron ions. Specifically, in the presence of physiologic quantities of iron ions, approximately 7 of every 1,000 DNA bases were modified. Addition of the superoxide anion scavenger, superoxide dismutase, the hydrogen peroxide scavenger, catalase, the hydroxyl scavenger, dimethylsulfoxide, or the iron chelator, deferoxamine, to DNA mixtures containing PMA, neutrophils, and iron ions, greatly decreased the yield of the damaged DNA base products. Our results indicate that stimulated human neutrophils can damage each of the four bases in DNA. It is likely that hydroxyl radical, generated via an iron catalyzed Haber-Weiss reaction, mediates neutrophil-induced DNA base damage, since: (a) the chemical structure of neutrophil-induced DNA base damage is consistent with a hydroxyl radical-mediated mechanism, (b) hydroxyl radical generated via ionizing radiation in aqueous solution produces DNA base modifications that are identical to neutrophil-induced DNA base modifications, (c) iron ions increase neutrophil-induced DNA base damage, and (d) iron chelators or scavengers of superoxide anion, hydrogen peroxide or hydroxyl radical decrease neutrophil-induced DNA base damage. PMID- 2553780 TI - Synovial procollagenase activation by human mast cell tryptase dependence upon matrix metalloproteinase 3 activation. AB - Mast cells have been implicated in the pathogenesis of the matrix degradation observed in the cartilaginous and osseous structures of the rheumatoid joint. We previously reported that human mast cell tryptase, a 134-kD granule-associated neutral protease, is present in rheumatoid synovium and can activate collagenase in crude culture medium in vitro. the present study attempts to depict the precise mechanism of this activation. To express full activation of latent collagenase, matrix metalloproteinase 3 (MMP-3) or stromelysin, can be activated by tryptase in a time and dose-dependent manner. Tryptase was not capable of generating active collagenase in the crude media from cultured rheumatoid synoviocytes depleted of proMMP-3 by immunoadsorption. In addition, the function of the tissue inhibitor of metalloproteinases (TIMP) was not altered by tryptase, and SDS-PAGE analysis revealed no degradation of TIMP by tryptase. The tryptase dependent activation of synoviocyte procollagenase thereby appears to be entirely dependent upon its ability to activate proMMP-3. PMID- 2553782 TI - Quantitative study of glutamic acid decarboxylase-immunoreactive neurons and cytochrome oxidase activity in normal and partially deafferented rat hindlimb somatosensory cortex. AB - Somatosensory cortex reorganizes following restricted deafferentation so that deprived neurons acquire new receptive fields. Electrophysiological data suggest that a decrease in inhibition might be one of the mechanisms contributing to these changes. This hypothesis was tested by evaluating quantitatively glutamic acid decarboxylase (GAD) immunoreactivity and cytochrome oxidase (CO) activity in normal and partially deafferented rat hindlimb somatosensory cortex. In normal animals, there were laminar differences in the frequencies of GAD+ cells that correlated with the levels of CO activity. Two weeks after transection of the sciatic nerve, CO levels were reduced in all layers of the hindlimb somatosensory cortex contralateral to the nerve transection whereas the frequencies of GAD+ cells were unchanged except in layer IV where a 16% decrease was observed. This observation is consistent with the hypothesis that the expression of GAD in layer IV is partially controlled by the amount of afferent input. The ability of novel inputs to develop stable patterns of excitation in deafferented somatosensory cortex may depend upon the reduction of GABAergic inhibition which follows deafferentation. PMID- 2553781 TI - Expression of the ErbA-beta class of thyroid hormone receptors is selectively lost in human colon carcinoma. AB - Members of the erbA gene family are involved both in control of differentiation and in neoplasia. V-erbA, a retroviral oncogene, blocks avian erythroid differentiation. V-erbA-related transcripts are physiologically expressed in multiple normal tissues. They encode a family of transcriptional regulatory factors, some of which are thyroid hormone receptors. In man, two genes, erbA alpha and erbA-beta, are transcriptionally active. We examined expression of erbA related transcripts in normal and neoplastic colon. In normal colon mucosa, as well as in a colon polyp and in a colon polyp cell line, three characteristic erbA-related transcripts were consistently found. One transcript of 6 kb was erbA beta related. Two transcripts of 2.7 and 5.2 kb were erbA-alpha related. In eight patients' colon carcinomas expression of the 6-kb erbA-beta transcript was absent or markedly diminished when compared with the same patients' noninvolved mucosa. In contrast, expression of the two erbA-alpha transcripts was the same in both colon carcinoma and noninvolved mucosa. No evidence was found of erbA-beta gene deletion in any of the tumors lacking erbA-beta expression. These data suggest that selective loss of normally present erbA-beta gene expression accompanies malignant transformation of the colonic epithelial cell. PMID- 2553783 TI - Gap junction protein in weakly electric fish (Gymnotide): immunohistochemical localization with emphasis on structures of the electrosensory system. AB - Electrotonic transmission via gap junctions appears to be essential for both the relay and integration of information in nuclear groups involved in the electrolocation and electrocommunication systems of weakly electric fish. An affinity-purified antibody against the 27 kD gap-junctional polypeptide (GJP) from rat liver was used to determine immunohistochemically the distribution of GJP-immunoreactivity (GJP-IR) in electrosensory structures and some other brain regions of the gymnotiform fish, Apteronotus leptorhynchus. At the ultrastructural level, immunolabelling with this antibody was localized, in part, to neuronal and glial gap junctions where it was assumed to recognize a junctional polypeptide. By light microscopy, the vast majority of immunoreactive elements appeared either as fine puncta or as varicosities along fibers that exhibited immunostained intervaricose segments. Diffuse immunoreactivity within cell bodies was rare, being most evident in giant relay neurons and presumptive glial cells within the pacemaker nucleus and in neurons within the posterior raphe nucleus. The distribution of punctate and fibrous GJP-IR was remarkably heterogeneous with respect to density; large areas of the forebrain and most major fiber tracts were nearly devoid of immunoreactivity, whereas concentrations of puncta delineating patches within the inferior lobe of the hypothalamus and the vagal sensory nucleus were so dense as to appear as uniform deposition of immunoperoxidase reaction product at low magnification. Some structures known to be associated with the electrosensory system, including the nucleus electrosensorius and nucleus praeeminentialis, were among the brain regions containing the highest concentrations of immunoreactivity. At the cellular level, expected patterns of GJP-IR were observed in the pacemaker nucleus, torus semicircularis, and electrosensory lateral line lobe. In each of these structures punctate immunoreactivity was seen in apposition to cell bodies or dendrites of neurons known to receive gap junction contacts. In addition, the dendrites of neurons within the prepacemaker nucleus were laden with a striking array of puncta, suggesting that interactions via gap junctions may be a significant feature of these neurons. These immunohistochemical results are consistent with previous electrophysiological and ultrastructural observations pointing to the importance of electrotonic communication in the electrosensory system of weakly electric fish, and suggest that gap junctions may also contribute to neural transmission in central nervous system related to other functions in these teleosts.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2553784 TI - Short-term keloid treatment in vivo with human interferon alfa-2b results in a selective and persistent normalization of keloidal fibroblast collagen, glycosaminoglycan, and collagenase production in vitro. AB - Two intralesional injections of interferon alfa-2b (1.5 million U per injection) into a progressively enlarging keloid resulted in a 41% reduction in its area. Fibroblasts cultured from the keloid before and 9 days after the initial injection were compared with fibroblasts cultured from the patient's normal skin with respect to proliferation and production of connective tissue matrix components and collagenase. There were no significant differences in the in vitro doubling times of keloidal fibroblasts before (p greater than 0.2) and after (p greater than 0.5) treatment with interferon alfa-2b, as well as of normal fibroblasts, in subconfluent cultures. Multiple passages of keloidal fibroblasts before interferon alfa-2b therapy assayed as confluent cultures produced more collagen (171%, 187%, and 204%), more glycosaminoglycans (153% and 141%), and less collagenase (26% and 31%) than the patient's own normal fibroblasts. In contrast, keloidal fibroblasts after interferon alfa-2b therapy persistently produced normal or subnormal amounts of collagen (107%, 73%, and 64%) and glycosaminoglycans (97% and 96%) and normalized levels of collagenase activity (96% and 86%). Normal amounts of fibronectin were produced by keloidal fibroblasts before and after treatment with interferon alfa-2b. PMID- 2553785 TI - Contact sensitivity to nonoxynols as a cause of intolerance to antiseptic preparations. AB - Twelve cases of allergic contact dermatitis caused by antiseptic preparations are presented. The reactions resulted not from the active principles but rather from nonoxynols used in the offending substances as nonionic surface-active agents. Nonoxynols are ethoxylated alkyl phenols or nonylphenylethers that conform in general to the formula C9H19C6H4(OCH2CH2)nOH. They have emulsifying, wetting, foaming, and solubilizing properties and are used in a large number of industrial, household, agricultural, cosmetic, and pharmaceutical products. They also are used as spermicides. There are very few reports in the literature of skin problems caused by nonoxynols. PMID- 2553786 TI - Clinically occult cytomegalovirus present in skin biopsy specimens in immunosuppressed hosts. AB - Three patients with acquired immunosuppression in whom skin biopsy specimens were obtained to aid in the diagnosis of varying cutaneous eruptions are described. Amphophilic, granular, intranuclear inclusions within endothelial cells compatible with the presence of cytomegalovirus were found unexpectedly. Clinically, occult cytomegalovirus in the skin may be more common than generally appreciated and may not be the sole cause of the eruption for which biopsy specimens are obtained. Also, the need to remain alert for polymicrobial cutaneous infections in the immunocompromised host is emphasized. PMID- 2553787 TI - Aquagenic pruritus: treatment with sodium bicarbonate and evidence for a seasonal form. PMID- 2553788 TI - Lymphomatoid papulosis and progression to T cell lymphoma: an immunophenotypic and genotypic analysis. AB - A 37-year-old white man had untreated lymphomatoid papulosis for 12 years before a submandibular T cell immunoblastic lymphoma developed. A genetic abnormality, composed of extra chromosomal material attached to the short arm of chromosome 9, was detected in the lymphoma tissue but not in the skin. The lymphomatoid papulosis skin lesions did not manifest clonal T cell receptor gene rearrangements, but the submandibular lymphoma tissue was clonal and of T cell lineage. The patient's lymphoma responded well to combination chemotherapy, but the lymphomatoid papulosis remains active. PMID- 2553789 TI - Prospective evaluation of thoracic-duct drainage in the treatment of respiratory failure complicating severe acute pancreatitis. AB - Thoracic duct drainage (TDD) may be of value for removing toxic substances released by the inflamed pancreas and which are responsible for lung damage. We have prospectively assessed the efficacy of TDD in improving pulmonary gas exchange in 12 patients with severe acute pancreatitis (SAP) complicated by persistent respiratory failure despite standard conservative treatment including peritoneal dialysis in 8 patients. In group A were 6 patients (mean Ranson score = 7.3) with adult respiratory distress syndrome (ARDS) and in group B were 6 hypoxemic patients (mean Ranson score = 6.6) judged to be at risk of developing ARDS. The duration of TDD ranged from 3 to 10 days and the total amount of drained lymph (L) varied from 770 to 15,600 ml. Immunoreactive trypsin levels were significantly higher in L when compared to blood in both groups. Leukocyte myeloperoxidases in L (normal value less than than 332 +/- 82 ng/ml in plasma) were increased in 5 of 5 group A patients (830 +/- 317 ng/ml) and in 3 of 6 patients in group B (671 +/- 467 ng/ml). After TDD pulmonary gas exchange as measured by median PaO2/FiO2 (mmHg) improved from 148 +/- 60 to 285 +/- 42 in group A and from 192 +/- 37 to 330 +/- 42 in group B (p less than 0.05). All patients were weaned after ventilation for a mean of 8 days in group A and 4 days in group B. All patients survived apart from 1 group B patient who died of sepsis on day 34.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553790 TI - Nutritional management of plasma lipid disorders. AB - Prevention of vascular disease and acute pancreatitis is the goal of hyperlipidemia treatment. The risk of coronary heart disease (CHD) increases with increasing plasma cholesterol levels because low-density lipoprotein (LDL), the major carrier of cholesterol in the plasma, is atherogenic. High-density lipoprotein (HDL), especially the HDL2 subfraction, protects against CHD. Hypertriglyceridemia, although not an independent risk factor for CHD, is generally accompanied by low HDL cholesterol (HDLch), which may predispose to CHD. Reducing plasma LDL and raising HDL levels are thus goals in preventing CHD. Serum LDL levels may be lowered by reducing saturated fat and cholesterol intake; weight loss may decrease LDL but is more effective in lowering plasma triglycerides and raising HDLch. The percent of total calories from polyunsaturated, monounsaturated, and saturated fats should be less than 10%, up to 10-15%, and less than 10%, respectively. High cholesterol intake increases the flux of cholesterol, which may be harmful to arterial walls, but beyond a certain point does not increase plasma cholesterol levels. Some diets change the composition rather than the level of LDL and apoproteins. Weight reduction and maintenance are the most effective dietary measures to lower plasma triglycerides; omega-3 fatty acids (fish oils) have shown promise in reducing triglyceride but not cholesterol levels. Substitution of starch for sugar lowered triglyceride levels toward normal in hypertriglyceridemia patients. Fasting triglyceride levels rise in all individuals fed high-carbohydrate diets, but the high levels persist in hypertriglyceridemia patients. Weight loss, cessation of cigarette smoking, increased physical activity, good control of diabetes, and moderate alcohol use all raise HDLch levels. Vitamin E deficiency causes neurological sequelae in children with severe malabsorption problems due to abetalipoproteinemia or cholestatic liver disease. PMID- 2553791 TI - Recent advances in carbohydrate nutrition and metabolism in diabetes mellitus. AB - The last decade has brought a tremendous turnaround in nutritional recommendations for diabetes management, moving from low-carbohydrate, high-fat diets to high-carbohydrate, high-fiber, low-fat diets. Clinical studies which furthered understanding of carbohydrate metabolism greatly contributed to the change. This paper reviews recent advances in selected aspects of carbohydrate nutrition and metabolism in diabetes mellitus. PMID- 2553792 TI - Nuclear enzymes, fragile sites, and cancer. AB - The finding of a large number of recurrent fragile sites on human chromosomes provides suggestive evidence for the origin of recurrent chromosomal rearrangements that are a common feature in most neoplasias. These fragile sites can be induced by a wide variety of mutagens and carcinogens that are known to act through different molecular mechanisms. They correlate with two-thirds of the recurrent cancer chromosomal breakpoints known thus far and with the location of most mapped oncogenes and growth factors. Because we have now found that a large number of fragile sites can also be induced by agents such as DNase I and benzo[a]pyrene diol-epoxide (BPDE), which are known to induce hypersensitive chromatin sites, it is possible that a class of fragile sites represents regulatory regions of active genes. We have also introduced restriction enzymes into cultured lymphocytes and found that fragile sites are expressed in a pattern similar to that produced by DNase I and BPDE. Because fragile sites have a higher expression with advancing age, cigarette smoking, and deficiency of folic acid and B12 vitamins, they may represent active genomic sites that are vulnerable to physiological and environmental disturbance. PMID- 2553793 TI - Hormonal regulation myometrial adrenergic responses: the receptor and beyond. AB - The contractile response of the uterus is modified by sex steroids. In rabbit uterus, oestrogen promotes alpha-adrenergically-mediated contraction, whilst progesterone treatment results in beta-adrenergic relaxation. Examination of the mechanisms responsible for these changing adrenergic responses with sex steroids reveals multiple sites of regulation. Oestrogen increases alpha 1-receptor concentration and the linkage of the receptor to phospholipase C. In addition to this direct effect to promote contraction, oestrogen also uncouples the beta receptor from adenylate cyclase. Progesterone, conversely, promotes relaxation through beta-receptors by uncoupling alpha 2-receptors from inhibition of adenylate cyclase. Thus sex steroids can regulate specific agonist responses at and beyond the receptor. PMID- 2553794 TI - Role of sympathetic neurons in development of beta-adrenergic control of ornithine decarboxylase activity in peripheral tissues: effects of neonatal 6 hydroxydopamine treatment. AB - Sympathetic neurons are thought to regulate the development of their postsynaptic targets. In the current study, we examined the effects of sympathectomy with 6 hydroxydopamine in neonatal rats on the ontogeny of beta-receptor binding sites and their linkage to both cyclic AMP production and ornithine decarboxylase activity. Cardiac norepinephrine levels and turnover were used to confirm the completeness and permanence of the lesion. The ability of isoproterenol, a beta adrenergic agonist, to stimulate ornithine decarboxylase (a growth-related enzyme) in heart, lung and kidney, was reduced by neonatal sympathectomy; the effect persisted into young adulthood. The effect represented a selective uncoupling of enzyme activity from receptor activation, as receptor binding capabilities were unaffected and the linkage of beta-receptors to cyclic AMP was enhanced. Comparison of the effects of peripheral sympathectomy with those of central lesions (intracisternal 6-hydroxydopamine) confirmed the importance of sympathetic nerve terminals in determining the coupling of the receptors to ornithine decarboxylase. These data suggest that sympathetic neurons program their target organs to specific trophic responses during development. PMID- 2553795 TI - Development of tolerance to ethanol-induced suppression of breathing movements and brain activity in the near-term fetal sheep during short-term maternal administration of ethanol. AB - The effect of short-term maternal ethanol administration on the ethanol-induced suppression of fetal breathing movements, electrocortical (ECoG) activity, and electroocular (EOG) activity was determined in the near-term fetal sheep. Twelve conscious instrumented pregnant ewes (between 125 and 139 days of gestation; term, 147 days) received 1-h intravenous infusion of 1 g ethanol/kg total body weight daily for six days (n = 6) or an equivalent volume of normal saline daily for six days (n = 6). On the seventh day, the ethanol- and saline-pretreated animals were administered 1 g ethanol/kg total body weight. A further six ewes received 1-h intravenous infusion of 1 g ethanol/kg total body weight (n = 3) or an equivalent volume of normal saline (n = 3) daily for thirteen days with both groups receiving 1 g ethanol/kg total body weight on day fourteen. Fetal ECoG and EOG activities, and fetal breathing movements were monitored continuously over the post- operative and experimental periods. Saline infusion had no significant effect on the parameters studied. Fetal breathing movements were suppressed for 8 h after the first ethanol dose, and were not significantly suppressed after fourteen days of once-daily, maternal ethanol administration. Low-voltage ECoG and EOG activities were suppressed for 3 h after the first ethanol dose, and were not significantly suppressed after seven days of repeated ethanol administration. Maternal and fetal blood gases and acid-base balance were not significantly affected by maternal ethanol administration. These data demonstrate that short term maternal administration of ethanol results in the development of tolerance to ethanol in the mature fetus. PMID- 2553796 TI - Vaccines against Japanese encephalitis. PMID- 2553797 TI - Susceptibility status of Culex vishnui to DDT and dieldrin in Deoria, Uttar Pradesh. PMID- 2553798 TI - Seroepidemiological study of Japanese encephalitis in Dimapur, Nagaland. AB - A seroepidemiological study of Japanese encephalitis (JE) in Dimapur, Nagaland was carried out following an outbreak of the disease between July, 1985 and February, 1986. Altogether 50 persons were affected with 30 (60 per cent) deaths. The attack and death rates per 1000 were more in Nagas viz. 0.55 and 0.34 than non-Nagas viz. 0.33 and 0.20 respectively. All ages and both sexes were affected. Of the nine mosquito species encountered Culex vishnui showed the highest density (44.5/MH). Culture of mosquito pool did not yield any viral agent. A total of 311 serum samples comprising 95 humans, 166 animals and 50 birds were tested for the presence of haemagglutination inhibition (HI) antibodies against Chikungunya and three flavirus antigens, viz. JE, WN and DEN-2. The overall flavivirus HI antibody in humans was positive in 26 (27.3 per cent) almost identical to JE antibody prevalence. The per cent positivity of HI antibodies to JE, WN and DEN-2 were 42.2 per cent, 22.2 per cent and 13.3 per cent in the affected and 14.2 per cent, 10 per cent and 6 per cent in the unaffected area. The activity of Alpha (Chikungunya) virus though very low was significantly higher (chi 2 = 5.57) in the affected area. The prevalence of JE antibody was 77.7 per cent in dogs, 52 per cent in cattle, 34 per cent in pigs and 21.1 per cent in goats. Of the five species of birds, flavivirus and JE antibodies were detected in 21.4 per cent pigeons and 22.2 per cent heron egrettes. Neutralisation test established the distinct role of JE virus over other related flavivirus antigens. PMID- 2553799 TI - Differential hormone-binding characteristics of thyroxine-binding globulin in hepatocellular carcinoma and cirrhosis. AB - To determine whether previous observations suggesting reduced ability of thyroxine-binding globulin (TBG) to bind exogenous thyroid hormones in hepatocellular carcinoma (HCC) might form the basis of a useful clinical test, we have investigated a larger series involving 49 patients and 10 healthy subjects. The binding ratio (mol hormone bound/mol TBG at a given hormone concentration) for both T4 and T3 was significantly reduced when compared to both cirrhotic and normal subjects. No overlap in T4 binding occurred between HCC patients and the control groups. Serial measurements on serum samples obtained from five cirrhotic patients who ultimately developed HCC revealed significant reductions in binding for both hormones up to 60 months prior to the clinical diagnosis of HCC. A reduced ability to bind thyroid hormones appears to be a specific feature of serum TBG from HCC patients and is one of the earliest alterations seen among cirrhotic subjects who ultimately develop the disease. PMID- 2553801 TI - Density gradient separation of two populations of lysosomes from rat parotid acinar cells. AB - Exocrine acinar cells possess two cytochemically distinct populations of secondary lysosomes. One population is Golgi associated and has demonstrable acid phosphatase (AcPase) activity, whereas the second is basally located and lacks AcPase activity but has trimetaphosphatase (TMPase) activity. The basal lysosomes are tubular in shape and rapidly label with horseradish peroxidase (HRP) after intravenous injection. In the present study using isolated rat parotid acinar cells, the two lysosomal populations were separated by cell fractionation on Percoll density gradients and were analyzed biochemically and by EM cytochemistry. On 35% Percoll gradients, two peaks of AcPase and beta hexosaminidase, both lysosomal marker enzymes, and succinic dehydrogenase, an enzyme marker for mitochondria, could be resolved. The major peaks of beta hexosaminidase and succinic dehydrogenase and the minor peak of AcPase corresponded with the dense lysosome fraction. The major peak of AcPase and the minor peaks for beta-hexosaminidase and succinic dehydrogenase coincided with the light membrane fraction. Galactosyl transferase (a marker enzyme for Golgi saccules) and 5'-nucleotidase (a plasma membrane marker) were also associated with this fraction. By electron microscopy, the light membrane fraction was seen to contain tubular elements, multivesicular bodies (MVB), Golgi saccules, GERL, immature secretory granules, and some mitochondria. Electron microscopic cytochemical examination showed that these tubular structures were lysosomes. The dense lysosome fraction contained lysosomes positive for both AcPase and TMPase. After continuous incubation of isolated acinar cells with HRP, reaction product was rapidly localized to the light membrane fraction (greater than 2 min), where it was found in vesicles and tubular lysosomes. By 10 min it was present in MVB and tubular lysosomes, but by 60 min no HRP reaction product had appeared in the dense lysosomes. These results demonstrate that the tubular lysosomes are separable from dense lysosomes, typical secondary lysosomes, and are involved in the initial stages of endocytosis. PMID- 2553802 TI - Testosterone immunoreactivity in the seminiferous epithelium of rat testis: effect of treatment with ethane dimethanesulfonate. AB - Androgens drive spermatogenesis by processes that are largely unknown. Direct effects on germ cells and indirect effects mediated via testicular somatic elements are currently under consideration, and specific localization of androgens in seminiferous tubules may provide information as regards this. Adult male rats were injected with ethane dimethanesulfonate (EDS; 75 mg/kg body weight) or vehicle. Testes were fixed and paraffin-embedded for localization of testosterone immunoreactivity 1 and 2 weeks after treatment, using the unlabeled antibody (PAP) technique. Plasma testosterone dropped from a pre-treatment level of 2.3 ng/ml to below 0.2 ng/ml 3 days after EDS injection and remained at low levels until the end of observation, accompanied by a progressive decrease in testicular weight. In the seminiferous tubules of vehicle-injected males, testosterone immunoreactivity was found in nuclei of spermatocytes and spermatids and in nuclei and the cytoplasm of Sertoli cells, and showed typical variations according to the stage of spermatogenesis. One week after EDS treatment, immunoreactivity had disappeared from the seminiferous epithelium. Two weeks after treatment, staining of germ cells was detected in two out of four males. The disappearance and reappearance of immunoreactivity coincided with the time course of EDS effects on rat Leydig cells, and we conclude that it corresponds to androgen specifically localized in fixed, paraffin-embedded tissue. Because staining of germ cell nuclei varied with the stage of spermatogenesis, the technique may detect a physiologically relevant androgen fraction; its location suggests that androgens may also directly affect certain germ cell stages. PMID- 2553800 TI - Immunohistochemical detection of 1,25-dihydroxyvitamin D3 receptors and estrogen receptors by monoclonal antibodies: comparison of four immunoperoxidase methods. AB - We developed an immunohistochemical method for visualization of vitamin D (VDR) and estrogen receptors (ER) in cryostat sections, using monoclonal antibodies (MAb) to the vitamin D receptor and estrogen receptor, respectively. This method is based on an avidin-biotin labeling technique (LAB). To establish a reliable and sensitive method which can be used easily as a routine diagnostic procedure, we systematically compared four different immunoenzymatic methods with respect to their efficiency in detecting vitamin D and estrogen receptors. Compared to the indirect bridged avidin-biotin (IBRAB), the peroxidase- anti-peroxidase (PAP), and the avidin-biotin complex (ABC) methods, the LAB method produced stronger staining intensities and had higher detection efficiency for both vitamin D and estrogen receptors. In addition, the LAB method had a higher spatial resolution compared to the ABC technique in detection of VDR in normal human skin biopsies. In the case of steroid receptors, i.e., nuclear antigens, immunohistochemistry must deal with a relatively low number of antigenic sites per cell, restricted accessibility of the antigens, and slight differences in antigen concentrations among cells. Under these particular conditions, the chemical properties of the conjugates used in the LAB method may explain why it is superior to the other methods. Consequently, the LAB method is recommended for visualization of ER and VDR. PMID- 2553803 TI - Ultrastructural localization of adenylate cyclase in chicken bone cells. AB - We investigated adenylate cyclase distribution in 6-day-and 3-week-old calvariae and in 6-day-old long bone metaphyses from chickens. Reaction product distribution was on the plasma membrane of osteoblasts, pre-osteoblasts, and forming osteocytes which contacted one another. Osteoclasts and mature osteocytes lacked reaction product. Six-day calvariae reacted less intensely than the other two tissues. In controls, reaction product was markedly diminished or eliminated by removal of forskolin or substrates, or by addition of the inhibitor 2',5' dideoxyadenosine. The results indicate the importance of cyclic AMP in osteoblast regulation. Osteocytes and osteoclasts may involve alternate mechanisms as major regulatory systems. PMID- 2553804 TI - Immunohistochemical localization of atrial natriuretic peptide receptor in bovine kidney and lung. AB - Receptors for atrial natriuretic peptide (ANP) were localized in the alveoli and bronchiolar smooth muscle cells of bovine lung and in podocytes of the kidney by immunofluorescence and immunoperoxidase methods. Two specific antisera were raised against the ANP receptor purified from bovine lung plasma membranes: anti Rc 140 and anti-Rc 70. Anti-Rc 140 was raised against the 140 KD native receptor having a homodimeric structure, and anti-Rc 70 was elicited by immunizing a rabbit with the 70 KD reduced subunits. Essentially identical staining patterns were obtained with both antisera. Identification of ANP receptor sites would provide useful information in understanding the pulmonary and renal actions of ANP. PMID- 2553805 TI - Concentration of poliovirus type I from cow milk using an adsorption-elution method. AB - Cow milk was experimentally contaminated with a vaccine strain of poliovirus type I. A concentration procedure was utilized to assay for the virus in a milk sample using adsorption on aluminium sulphate at pH 4.5-5.5, aluminium sulphate concentration being 0.15 g. 1(-1), followed by elution with 0.1 M Na2HPO4 pH 9.5 and subsequent detection in cultured VERO cells (kidney cells from the monkey Cercopithecus aethiops). The yield of the virus ranged from 14 to 58% of the inoculated amount, the mean value being 27%. PMID- 2553806 TI - Cardiovascular, renal and endocrine effects of alpha-human atrial natriuretic peptide in patients with Cushing's syndrome and primary aldosteronism. AB - The pharmacological effects of synthetic alpha-human atrial natriuretic peptide (alpha-hANP) in patients with Cushing's syndrome and primary aldosteronism were compared with those in normal volunteers. An infusion of synthetic alpha-hANP at 0.1 microgram/kg per min for 20 min produced a maximal plasma hANP level of 800 1200 pg/ml in patients with Cushing's syndrome and primary aldosteronism, and in normal subjects. There were significant decreases in the mean blood pressure (-10 to -15 mmHg) in patients with Cushing's syndrome and primary aldosteronism, similar to those in normal subjects. The plasma cyclic 3'5'-guanosine monophosphate (cGMP) concentrations of both groups of patients were increased fivefold over the baseline level following the infusion. Infusion of synthetic alpha-hANP caused a greater increase in the rate of sodium excretion in patients with Cushing's syndrome and primary aldosteronism compared with normal volunteers. The plasma cortisol and aldosterone concentrations did not, however, significantly change during alpha-hANP infusion in either the patients with Cushing's syndrome or those with primary aldosteronism. As synthetic alpha-hANP has a potent hypotensive effect in hypertensive patients with Cushing's syndrome and primary aldosteronism, a significant reduction in blood pressure and natriuresis seems to occur without affecting adrenocortical steroidogenesis. PMID- 2553807 TI - Heterogeneous distribution and transmembrane signaling properties of lymphocyte function-associated antigen (LFA-1) in human lymphocyte subsets. AB - The role of LFA-1, a member of the integrin supergene family, in intercellular adhesion, including lymphocyte-endothelial cell (EC) binding, has been established. We now demonstrate that differences in LFA-1 cell surface density are responsible for the variable adhesion efficiency of lymphocyte subsets to EC. Electrophoretic analysis revealed multiple glycosylated isoforms of both alpha and beta subunits, largely as a result of different degrees of sialylation, with variable expression among different lymphocyte subsets. Neuraminidase digestion before EC adhesion increased the binding efficiency of all lymphocyte subsets, although the relative increase in each subset was proportional to the initial LFA 1 sialic acid content. LFA-1 cross-linking resulted in phosphoinositide hydrolysis and a rise in [Ca2+]i when using anti-alpha but not anti-beta subunit antibodies. These findings indicate that the density of LFA-1 on lymphocyte subsets controls their adhesive properties, and that the LFA-1 alpha subunit has transmembrane signaling properties that may result in activation events after interaction with its natural ligands. PMID- 2553808 TI - Biochemical characterization of the fibronectin binding sites for IgG. AB - Plasma fibronectin (Fn) is a constituent of cryoglobulins and has been shown to interact with immune complexes. In a previous report we demonstrated that Fn specifically bound to IgG immobilized on a solid matrix. To localize and biochemically characterize the sites on the Fn molecule involved in this interaction, Fn was enzymatically cleaved with subtilisin and subjected to IgG affinity chromatography. Three major polypeptide fragments of 16 kDa, 22 kDa, and a triplet of 26- to 29-kDa bound IgG. They were localized to three separate regions of the molecule by Western blot analysis using antisera to specific regions of the Fn molecule, by amino acid sequencing, and by their previously described heparin binding affinities. The 22-kDa fragment interacted with IgG under physiologic conditions and it is localized at the N-terminal of the Fn molecule. The 16-kDa and 26- to 29-kDa fragments bound to IgG under conditions of lower ionic strength; the former commences at residue 588, carboxyl-terminal to the collagen binding region and the latter begins at residue 1597, carboxyl terminal to the cell binding domain. The interaction of Fn with Ig has significant implications in host defense and also in immune complex disease where basement membrane Fn may sequester immune complexes from the circulation. PMID- 2553809 TI - Natural killer cell and granulocyte Fc gamma receptor III (CD16) differ in membrane anchor and signal transduction. AB - CD16 is a low affinity Fc gamma R III expressed on granulocytes, macrophages and large granular lymphocytes, the mediators of antibody-dependent cellular cytotoxicity and NK. The occupancy of CD16 by aggregated IgG on large granular lymphocytes induces expression of activation markers, release of inflammatory mediators and triggering of effector functions such as antibody-dependent cellular cytotoxicity. Recently we and others described that CD16 is anchored to the membrane of granulocytes via a phosphatidylinositol glycan moiety. Here we show that the CD16 molecule expressed on NK cells, cultured monocytes, and lung macrophages is not phosphatidylinositol glycan moiety anchored. It is not released with phosphatidylinositol-specific phospholipase C, and after removal of N-linked carbohydrate is 5 to 7 kDa larger than the granulocyte CD16 molecule, strongly suggesting the presence of transmembrane and cytoplasmic protein domains. Redirected killing of hybridoma targets expressing anti-CD16 surface Ig shows that NK cell CD16 is unable to do so. These findings demonstrate that NK cell and granulocyte CD16 have different membrane anchors and indicate that the type of membrane anchor is an important biologic mechanism for regulating the functional capacity of surface receptors. PMID- 2553810 TI - Migration of neutrophils across endothelial monolayers is stimulated by treatment of the monolayers with interleukin-1 or tumor necrosis factor-alpha. AB - To study the effects of the cytokines IL-1 and TNF-alpha on the transendothelial migration of neutrophils, human umbilical vein endothelial cells (HUVEC) were grown to confluence on connective tissue prepared from human amniotic membrane. Pretreatment of HUVEC-amnion cultures with rIL-1 beta (7.5 ng/ml) or rTNF-alpha (5 ng/ml) for 4 h resulted in rapid migration of from 20 to 50% of subsequently added neutrophils across the endothelial monolayer. In contrast, only 3 +/- 3% of added neutrophils penetrated the HUVEC monolayer in the absence of any stimulus. The number of neutrophils that migrated across cytokine-treated HUVEC was similar to the number that traversed untreated monolayers in response to gradients of FMLP; in addition, it was only 35% less than the number of neutrophils that migrated in response to leukotriene B4. No consistent additive effect was seen when migration was induced by both cytokine pretreatment of the HUVEC and a chemotactic gradient. The number of neutrophils that migrated across IL-1-treated cultures was proportional to the number added over the range of 2.5 x 10(5) to 4 x 10(6) neutrophils. When used at optimal concentrations, IL-1 and TNF-alpha were equally effective in stimulating neutrophil migration; no additive effect was seen when HUVEC were pretreated with optimal doses of both cytokines together. Direct addition of IL-1 or TNF-alpha to a 1-h migration assay had no effect on neutrophil adhesion to or migration across HUVEC, either in the presence or absence of a chemotactic gradient. Stimulation of neutrophil transendothelial migration in this system did not appear to be caused by adsorption of cytokine by the amniotic tissue, nor was it due to contamination of the cytokine preparations by LPS. These results suggest that IL-1 and TNF-alpha, generated at sites of inflammation, may act upon the endothelium to promote emigration of neutrophils from the vasculature. PMID- 2553811 TI - Function and regulation of the neutrophil MEL-14 antigen in vivo: comparison with LFA-1 and MAC-1. AB - The CD11/18 (LFA-1, Mac-1) molecules participate in neutrophil adhesion to cultured endothelium in vitro and are critical for effective neutrophil localization into inflamed tissues in vivo. More recently, the MEL-14 Ag, which was first defined as a lymphocyte homing receptor, has also been implicated in inflammatory neutrophil extravasation. Here we compare the regulation and function of these adhesion molecules on neutrophils during the in vivo inflammatory response. The MEL-14 Ag is expressed at high levels on bone marrow and peripheral blood neutrophils, but is lost on neutrophils isolated from the thioglycollate-inflamed peritoneal cavity. In contrast, Mac-1 is up-regulated on inflammatory neutrophils and little change is seen in the level of LFA-1 expression. In vitro activation of bone marrow neutrophils with PMA or leukotriene B4 results in a dose dependent increase in Mac-1 and decrease in MEL 14 Ag expression within 1 h after treatment, thus reflecting what is found during inflammation in vivo. Neutrophils activated in vitro or in vivo (MEL-14Low, Mac 1Hi) do not home to inflammatory sites in vivo, correlating with the loss of the MEL-14 Ag and the increased Mac-1 expression. Anti-LFA-1, anti-Mac-1, or MEL-14 antibody given i.v. suppress neutrophil accumulation within the inflamed peritoneum (38%, 30%, and 37% of medium control, respectively) without affecting the levels of circulating neutrophils. However, when FITC-labeled cells are precoated with the mAb and injected i.v., only MEL-14 inhibits extravasation into the inflamed peritoneum (25% of medium control). Finally, in ex vivo adhesion assays of neutrophil binding to high endothelial venules in inflamed-lymph node frozen sections MEL-14 inhibits greater than 90%. anti-LFA-1 20 to 30% and anti Mac-1 less than 10% of the binding of bone marrow neutrophils to inflamed-lymph node high endothelial venules. These results confirm that both the MEL-14 antigen and Mac-1/LFA-1 are important in neutrophil localization to inflamed sites in vivo, but suggest that their roles in endothelial cell interactions are distinct. PMID- 2553812 TI - Cytomegalovirus infection stimulates expression of monocyte-associated mediator genes. AB - Monocytes and tissue macrophages play important roles in host defense against virus infections and, in the case of human cytomegalovirus (HCMV) and HIV, may also be the reservoir for latent disease. Because these cells can also rapidly respond to most infections by secretion of inflammatory mediators, we were interested in determining if HCMV infection could have a direct activating effect on macrophage cytokine production. To do this, we primarily investigated the influence of HCMV infection on IL-1 beta-mRNA expression in peripheral blood monocytes and the promyelocytic cell line, ML-3 as well as the inflammatory response genes TNF-alpha, MAD-9, MAD-6, and MAD-2 in the promyelocytic ML-3 cell line. Exposure of ML-3 cells to the virus prior to induction of differentiation had little influence on mediator gene expression. However, induction of the macrophage phenotype by pretreatment of ML-3 cells with the phorbol ester, PMA, followed by HCMV challenge, resulted in a greatly extended period of expression of IL-1 beta, TNF-alpha, MAD-9, and CSF-1 but not MAD-6 and MAD-2. Constitutively expressed genes such as lysozyme and actin were not similarly modulated. Both RNA dot-blot and in situ hybridization studies demonstrated that infection of human peripheral blood monocytes with HCMV leads to sustained expression of IL-1 beta mRNA for up to 96 h, which contrasted markedly with mock-infected or LPS stimulated monocytes. Flow cytometric analysis of the intracellular levels of IL 1 beta protein in ML-3 cells indicated that not only was there more protein produced in infected cells, but that the majority of the cells had responded. Enhanced levels of the intracellular form of IL-1 beta in monocytes was confirmed by Western blot analysis. Cotransfection experiments were performed using IL-1 beta-CAT chimeric plasmids together with plasmids encoding HCMV-immediate-early gene region products. Transactivation of the IL-1 beta gene by region 2 of the immediate-early gene was observed in ML-3 cells that had been induced to differentiate prior to transfection. No stimulation of IL-1 beta promoter activity was observed in ML-3 cells that were undifferentiated prior to transfection. In summary, HCMV infection, although not leading to productive infection, nonetheless may contribute to the pathology of the infection through enhancement of monocyte inflammatory mediator gene expression with subsequent stimulation of protein synthesis. PMID- 2553813 TI - Specificity of peptide binding by the HLA-A2.1 molecule. AB - The HLA-A2 molecule contains a putative peptide binding site that is bounded by two alpha-helices and a beta-pleated sheet floor. Previous studies have demonstrated that the influenza virus matrix peptide M1 55-73 can sensitize target cells for lysis by HLA-A2.1-restricted virus-immune CTL and can induce CTL that can lyse virus-infected target cells. To assess the specificity of peptide binding by the HLA-A2.1 molecule, we examined the ability of seven variant M1 peptides to be recognized by a panel of M1 55-73 peptide-specific HLA-A2.1 restricted CTL lines. The results demonstrate that five out of the seven variant M1 55-73 peptides could be recognized by A2.1-restricted M1 55-73 peptide specific CTL lines. The two variant peptides that were not recognized by any CTL could bind to HLA-A2.1 as indicated by their ability to compete for presentation of the M1 55-73 peptide. In addition, 5 of a panel of 24 unrelated peptides tested could also compete for M1 55-73 presentation by HLA-A2.1. One peptide derived from the sequence of a rotavirus protein could sensitize HLA-A2.1+ targets for lysis by M1 55-73 peptide-specific CTL. We conclude from these studies that: 1) the HLA-A2.1 molecule can bind a broad spectrum of peptides; 2) T cells selected for the ability to recognize one peptide plus a class I molecule can actually recognize an unrelated peptide presented by that same class I molecule; and 3) a stretch of three adjacent hydrophobic amino acids may be an important common feature of peptides that can bind to HLA-A2.1. PMID- 2553814 TI - Synergistic effects of phorbol ester and INF-gamma on the induction of indoleamine 2,3-dioxygenase in THP-1 monocytic leukemia cells. AB - Indoleamine 2,3-dioxygenase (IDO) is a flavin-dependent enzyme which uses superoxide anion as a cosubstrate to catalyze the decyclization of the pyrrole ring of L-tryptophan to form formylkynurenine. This enzyme is induced in some tumor cells after treatment with IFN-gamma. The mechanism of induction of IDO in tumor cells by IFN-gamma was studied in THP-1 human monocytic leukemia cells. Before the addition of IFN-gamma, no IDO could be detected in these cells. Treatment of THP-1 cells with IFN-gamma produced an induction of IDO, with peak activity occurring 72 to 96 h after addition of IFN-gamma. Because phorbol esters are known to induce many enzymes in cells, most likely through the activation of protein kinase C, the effects of PMA on the induction of IDO were determined. PMA potentiated the IFN-gamma-induced elevation of IDO, but by itself, was unable to induce enzyme activity. Maximum induction of IDO in the presence of PMA and IFN gamma was obtained by preexposure of the cells to PMA for 48 h before the addition of IFN-gamma. Maximum induction of IDO after the addition of IFN-gamma occurred 24 to 48 h after addition of the cytokine to the culture medium. However, the induction of IDO does not appear to be potentiated through the activation of protein kinase C, because the addition of the protein kinase C inhibitor H-7 had no effect on the induction of IDO when the cells were exposed to PMA and IFN-gamma. Moreover, diacylglycerol was unable to replace PMA in these studies. Studies with cAMP and cGMP analogs suggest a role for these compounds in the regulation of IDO expression. PMID- 2553815 TI - Generation of biologically active C-reactive protein peptides by a neutral protease on the membrane of phorbol myristate acetate-stimulated neutrophils. AB - The association of human C-reactive protein (CRP) with nonstimulated and PMA stimulated human neutrophils and the concomitant degradation of CRP (monitored by TCA-soluble peptides and SDS-PAGE analysis) has been studied. Maximum association of 125I-labeled CRP with neutrophils and 125I-labeled CRP degradation during association with these cells was achieved by stimulating the neutrophils with PMA at 10 ng/ml; a concentration in which azurophil granule release was not significant. For PMA-stimulated neutrophils, the association of 125I-labeled CRP was 1.8 times higher and PMA-stimulated neutrophil-mediated degradation of the ligand was three times faster than that for nonstimulated cells. The neutrophil associated 125I-labeled CRP in the absence and presence of PMA proved on SDS-PAGE analysis to be approximately 50% degraded. There was a positive correlation between the extent of CRP degradation and the association of 125I-labeled CRP with neutrophils. In addition to generation of neutrophil associated CRP intermediates, small soluble CRP peptides were generated during association of CRP with neutrophils. These peptides inhibited superoxide production from opsonized zymosan-activated neutrophils by approximately 40% at 10 micrograms/ml. 125I-labeled CRP degradation mediated by nonstimulated neutrophils, and neutrophil-conditioned medium (from both non-stimulated and PMA-stimulated cells) was inhibitable by alpha 1-antitrypsin and approximately seven times less at 1 h than that occurring during 125I-labeled CRP-association with PMA-stimulated neutrophils. The degradation of 125I-labeled CRP mediated by PMA-stimulated neutrophils was not fully inhibitable by alpha 1-antitrypsin. The data point to the involvement of a membrane-associated serine protease, which is maximally activated by PMA, in the degradation of 125I-labeled CRP during association with neutrophils. Our results indicate that at an inflammatory site CRP-derived peptides can be produced that inhibit the action of activated neutrophils. PMID- 2553816 TI - Treatment of experimental disseminated Mycobacterium avium complex infection in mice with recombinant IL-2 and tumor necrosis factor. AB - Mycobacterium avium complex (MAC) is the most common bloodstream pathogen isolated from patients with AIDS. We have previously shown that TNF alone or in combination with IL-2 can activate human and murine macrophages in vitro to kill MAC strains isolated from disseminated infections. To determine whether treatment with TNF and IL-2 could effect the course of disseminated MAC infections in a murine model of disseminated MAC infection, we infected C57BL mice with 3 x 10(8) bacteria i.v. and 1 wk later administered: 1) IL-2, 100 micrograms/kg; 2) TNF, 25 micrograms/kg; 3) IL-2, 50 micrograms/kg, and TNF, 12.5 micrograms/kg; and 4) saline. IL-2 was injected i.p. daily with TNF being administered in cycles of 3 out of 4 consecutive days. Fourteen days after starting therapy, blood was cultured and mice were sacrificed for quantitative cultures of liver and spleen homogenates. IL-2, TNF, and IL-2/TNF treated groups showed an 87 +/- 5%, 57 +/- 9%, 88 +/- 6% decrease in bacteremia (p = 0.05 for TNF-treated animals and less than 0.04 for the other two groups, compared with control). The combination IL 2/TNF was the only treatment that showed a trend toward an absolute decrease in the number of bacteria in the blood. Reduction in colony counts of liver and spleen were 77 +/- 4% and 87 +/- 6%, respectively, for treatment with IL-2, 58 +/ 7% and 87 +/- 5% for TNF, and 60 +/- 10% and 82 +/- 6% for IL-2/TNF, respectively. These results suggest that both cytokines may play a role in the control of Mycobacterium avium infection and that the combination of a half-dose of IL-2 and TNF, despite not showing any greater efficacy, can be less toxic than TNF or IL-2 alone and might be useful for the therapy of disseminated infection. PMID- 2553817 TI - Analysis of foot-and-mouth disease virus-neutralizing idiotypes from immune bovine and swine with anti-murine idiotype antibody probes. AB - Rabbit anti-idiotypic antibodies (a-IdAb) induced by foot-and-mouth disease virus (FMDV) neutralizing mAb were used as probes to identify anti-FMDV Id in immune serum from bovine and swine. In a competitive RIA, at least two of the a-IdAb exhibited a dose-dependent capacity to compete with labeled virus for anti-FMDV antibodies from a convalescent bovine serum. These a-IdAb were immobilized on activated Sepharose and used to isolate anti-viral Id from bovine, swine, and murine FMDV immune sera. Both the bovine and swine antibodies recovered from the a-IdAb/Sepharose columns reacted with virus, and to a lesser extent with corresponding mAb-resistant virus variants. The binding of affinity isolated bovine and swine antibodies to virus was specifically inhibited by the homologous a-IdAb, and in addition, both were capable of neutralizing FMDV in suckling mouse protection and plaque reduction neutralization assays. Therefore, by means of a IdAb probes generated against FMDV murine Id, two neutralizing Id were identified in bovine and swine. These results suggest that FMDV-neutralizing epitopes recognized by murine systems play a role in the overall immunity of foot-and mouth disease-susceptible animals. PMID- 2553818 TI - A comparison of methods for the detection of human papillomavirus DNA by in situ hybridization with biotinylated probes on human carcinoma cell lines. Application to wart sections. AB - We compared nine different techniques for the detection of biotinylated DNA-DNA HPV hybrids on HeLa cells with 10-50 copies of HPV 18 DNA per cell. CaSki cells with 600 copies of HPV 16 DNA per cell and tissue sections from frozen or paraffin-embedded biopsy specimens. The cell samples were either cell deposits or cytocentrifuged or cultured slides. In most cases, the samples (cell deposits and tissue sections) were denatured with hybridization mixture prepared under stringent conditions (Tm = -17 degrees C) containing biotinylated DNA probes (cloned HPV types 1, 2, 6, 11, 16 and 18), at 90 degrees C for 10 min. In other cases (cytocentrifuged or cultured cells), the denaturation was performed by HCl hydrolysis and mild heating at 50 degrees C; the probes were denatured separately by heating. All the samples were further incubated overnight at 37 degrees C. For HPV DNA detection, three amplification levels were used on cell deposits. Only the techniques involving a three-step reaction (a rabbit anti-biotin antibody - a biotinylated goat anti-rabbit antibody - a complex of streptavidin-alkaline phosphatase or streptavidin-gold or streptavidin-fluorescein) gave satisfactory results, on both cell lines. With the one step reaction (an avidin-horseradish peroxidase, or streptavidin-alkaline phosphatase or streptavidin-fluorescein complex), no labeling of HeLa cells was observed with any of the HPV probes, including HPV 18. The techniques involving four steps (avidin or streptavidin - anti-avidin goat antibody or anti-streptavidin rabbit antibody - a biotinylated anti-goat (or anti-rabbit) antibody - a complex of avidin-biotin-peroxidase or streptavidin-biotin-alkaline phosphatase or streptavidin-biotin-horseradish peroxidase) resulted in high background on both cell lines. For the reproducible detection of low copy number of HPV DNA (less than 50 copies) such as occur in HeLa cells our data suggested that the three-step technique with the streptavidin alkaline phosphatase complex was the method of choice. The most intense labeling was always obtained with cell deposits and the technique was successfully applied to frozen and paraffin-embedded tissue sections from typical warts. PMID- 2553819 TI - Comparison of a liquid-phase blocking sandwich ELISA and a serum neutralization test to evaluate immunity in potency tests of foot-and-mouth disease vaccines. AB - Sera from cattle vaccinated against either foot-and-mouth disease virus (FMDV) strains A10 Holland, O1 BFS, or C1 Detmold were tested in a serum neutralization test (SNT) and a liquid-phase blocking sandwich ELISA (LBE), and the titers were compared with the results of intradermolingual challenge tests. The LBE test results were significantly more reproducible (P less than 0.005) than the SNT results. The correlation coefficients between SNT and LBE were 0.91 for FMDV strains A10 Holland and O1 BFS, and 0.82 for FMDV strain C1 Detmold (P less than 0.0005). The regression coefficient for strain A10 Holland was 0.80, for strain O1 BFS the value was 0.87, and for strain C1 Detmold it was 0.64. In probit analysis, titers at which 95% of the cattle were protected against challenge with the homologous strain were determined for the SNT and the LBE. In the SNT the 95% protection levels for strains A10 Holland were greater than or equal to 0.84, for O1 BFS greater than or equal to 1.59, and for C1 Detmold greater than or equal to 0.83. In the LBE they were greater than or equal to 1.28, greater than or equal to 1.71, and greater than or equal to 1.74, respectively. Because the SNT and the LBE are highly significantly correlated, and the LBE is more reproducible, the LBE is likely to predict protection more reliably than the SNT. PMID- 2553820 TI - The influence of different adjuvants on the immune response to a synthetic peptide comprising amino acid residues 9-21 of herpes simplex virus type 1 glycoprotein D. AB - The immuno-modulating properties of different adjuvant systems on the murine humoral and cellular immune response to a synthetic peptide comprising amino acid residues 9-21 of glycoprotein D of herpes simplex virus type 1 (HSV-1) were investigated. For immunization, the peptide was conjugated to ovalbumin or bovine serum albumin by glutaraldehyde and the adjuvants used in this study were Freund's complete adjuvant (FCA), aluminium hydroxide, the Ribi adjuvant system (RAS) and two non-ionic block polymer surfactants, viz. L101 and 31R1, in oil in water emulsions. High anti-peptide antibody titers were obtained after immunization with FCA, aluminium hydroxide, RAS and L101. All adjuvants, except RAS, stimulated the induction of delayed type hypersensitivity obtained after immunization with peptide 9-21 coupled to ovalbumin and elicited by injection of purified HSV-1 virions in the footpad. Challenge with a lethal dose of HSV-1 showed that mice immunized with peptide 9-21 coupled to ovalbumin in combination with FCA, RAS and L101, respectively, were significantly protected. Although immunization with peptide 9-21 coupled to ovalbumin combined with aluminium hydroxide stimulated induction of delayed type hypersensitivity, no significant protective immunity against the challenge was generated. PMID- 2553821 TI - Poliovaccine virus in the cerebrospinal fluid after oral polio vaccination. AB - In February-March 1985 an oral poliovirus vaccine campaign was launched in Finland in a population vaccinated earlier with inactivated poliovaccine. During this campaign a strain of poliovirus was isolated from the cerebrospinal fluid (CSF) of a 7-year-old girl 34 days after she had received oral poliovirus vaccine. She had long-lasting headache, vomiting and fever but no paralysis. This case demonstrates that poliovaccine virus can invade the central nervous system even after a complete course of inactivated poliovirus vaccine if the inactivated vaccine has been poorly antigenic against one of the three types of virus. PMID- 2553822 TI - Cell lines from Culicoides variipennis (Diptera: Ceratopogonidae) support replication of bluetongue virus. AB - Cell lines have been developed from 2-day-old embryos of the biting midge, Culicoides variipennis (Diptera: Ceratopogonidae). In North America C. variipennis is the primary insect vector of bluetongue virus (BTV), an orbivirus that causes disease of ruminants. The C. variipennis (CuVa) cells, grown in Schneider's Drosophila medium, consist primarily of a fibroblast-like cell type. CuVa cells are very hardy. They can grow over a wide range of temperature and pH and adapt to growth in minimal essential medium. BTV replicates to high titer (7.5-8.0 log10 50% tissue culture infectious doses/ml) in CuVa cells over a wide range of temperatures (19 degrees to 37 degrees C) without inducing any significant cytopathic effects. The highest BTV titers were obtained in CuVa cells grown at 25 degrees and 32 degrees C. Cells from C. variipennis can be useful for many diverse investigations. PMID- 2553823 TI - Immunohistochemical detection of an immediate early antigen of human cytomegalovirus in normal tissues. AB - As a member of the herpesvirus family, human cytomegalovirus (HCMV) induces a life-long latent infection in most individuals infected with it. This latent infection is subject to periodic reactivations that serve as an important source of disease and death in patients with defective immunologic responses. The specific types of cells that harbor latent HCMV have not yet been identified. To detect latent HCMV and identify the host cells carrying it, tissue sections from nine HCMV-seropositive normal individuals were examined using a murine monoclonal antibody specific for one of the immediate early (IE) antigens of HCMV. Cells expressing the IE antigen were detected in several different tissues from six of the subjects. Tissues found to contain positively stained cells included brain, kidney, spleen, lung, and liver. No positively stained cells were found in any tissue from seven HCMV-seronegative individuals studied by the same procedures. Cells undergoing productive infection with HCMV could not be detected in any of the tissues containing IE antigen-positive cells. It is proposed that these IE antigen-expressing cells represent at least some of the sites of HCMV latency in normal seropositive individuals. PMID- 2553824 TI - Cytomegalovirus hepatitis in liver transplantation: prospective analysis of 93 consecutive orthotopic liver transplantations. AB - Ninety-three consecutive orthotopic liver transplantations in 78 patients were followed prospectively to study the incidence of cytomegalovirus (CMV) hepatitis. CMV hepatitis occurred in 13 (17%). The diagnosis was established by both histology and culture in 5, only by histology in 6, and only by culture in 2. All 13 patients had CMV viruria and 9 had viremia at diagnosis. CMV hepatitis developed in 64% of CMV-seronegative (pretransplantation) patients who received a liver from a CMV-seropositive donor, compared with 3% or 6% of CMV-seropositive patients who received a liver from a CMV-seronegative or CMV-seropositive donor, respectively (P less than .001). CMV hepatitis was not a cause of fulminant or irreversible liver dysfunction in any of the 13 cases. Ganciclovir was administered to 6 of the 13 patients and was associated with clinical and virologic cure in 5. CMV hepatitis was self-limited in patients not treated with ganciclovir (illness less severe). The presence of inclusions within the liver tissue correlated with active disease. PMID- 2553825 TI - Neonatal antibody-dependent cellular cytotoxic antibody levels are associated with the clinical presentation of neonatal herpes simplex virus infection. AB - The role of antiviral antibodies in protection against neonatal herpes simplex virus (HSV) infection remains controversial. The relationship between neonatal and maternal anti-HSV antibodies and disease presentation was analyzed in 47 babies. Of the neonates, 77% had localized and 23% had disseminated HSV infection. Antibody-dependent cellular cytotoxic (ADCC) antibodies were evaluated in comparison with HSV neutralizing antibodies. High maternal (greater than 1:10(4)) or neonatal (greater than 1:10(3)) anti-HSV ADCC antibody levels or high neonatal antiviral neutralizing levels (greater than 1:20) were independently associated with an absence of disseminated HSV infection. Cochran-Mantel-Haenszel analysis demonstrated that ADCC levels were associated with disease status (P less than .02) while controlling for the level of neutralizing antibody. PMID- 2553827 TI - Immune response of patients with exanthema subitum to human herpesvirus type 6 (HHV-6) polypeptides. PMID- 2553826 TI - Rotavirus-specific cytotoxic T lymphocyte response of mice after oral inoculation with candidate rotavirus vaccine strains RRV or WC3. AB - The immunologic basis of homotypic and heterotypic protection by immunization with two candidate rotavirus vaccine strains (simian strain RRV and bovine strain WC3, respectively) was investigated. Mice were orally inoculated with RRV, and 6 d later splenic lymphocytes lysed target cells infected with RRV (serotype 3) but not with human rotavirus serotypes 1 or 2; 4 w after inoculation, cytotoxic T lymphocytes (CTLs) generated in vitro from CTL precursors were also strain specific. Also, the frequency of RRV-specific CTLs from CTLp after RRV immunization was 20-fold greater than that of cross-reactive CTLs. Inoculation of mice with WC3 (serotype 6), on the other hand, produced cross-reactive CTLs from CTLp at 4 w; CTLs lysed target cells infected with WC3 or human serotypes 1, 2, or 3 to the same extent. The frequency of cross-reactive CTLs after WC3 immunization was greater than or equal to 20-fold greater than that of WC3 specific CTLs. Cross-reactive, rotavirus-specific CTLs induced after WC3 immunization may in part explain the immunologic basis of protection against heterotypic challenge. PMID- 2553828 TI - Adenovirus enteritis in SIV-infected rhesus monkeys. PMID- 2553829 TI - Anti-interferon antibodies to interferon-alpha 2b: results of comparative assays and clinical perspective. AB - Previous studies have reported a low (less than 3%) incidence of anti-interferon (IFN) serum neutralizing antibodies following treatment with IFN-alpha 2b. Since this result contrasts with a higher incidence reported with IFN-alpha 2a, the question has been raised whether differences in assay techniques and patient comparability rather than inherent differences in the molecules might account for the reported differences in antibody incidence. In this report two patient groups, 151 hairy cell leukemia (HCL) patients and 101 patients with other malignancies, who have received long-term dosing with IFN-alpha 2b, are reported. The sera of both groups were studied before, during and after treatment by various assay methodologies. Utilizing three assay techniques, a less than 3% overall incidence of serum antibody formation was confirmed in these retested samples. With over 575 samples tested in multiple assays, the radioimmunoassay, as utilized in prior reports, demonstrated 99% agreement with a bioassay. Therefore, prior speculation that the assay technique for IFN-alpha 2b might produce a high false-negative rate was disproven. Additionally, the clinical outcome of these patients also failed to demonstrate a pattern of clinical relapse. In summary, these new analyses confirm the low (less than 3%) incidence of neutralizing antibody development following treatment with IFN-alpha 2b and confirm that no high rates of clinical relapse have developed in patients treated with chronic long-term dosing. Assay methodology does not appear to be a likely explanation for the low incidence of antibody formation reported with IFN-alpha 2. Rather, the unique molecular structure and pharmaceutical formulation of IFN alpha 2b remains the most likely explanation of its minimal antigenicity. PMID- 2553830 TI - Detection and incidence of neutralizing antibodies to interferon-alpha-n1. AB - Sera obtained from 612 patients enrolled in 40 different clinical trials with an interferon-alpha-n1 (IFN-alpha-n1) preparation have been tested for neutralizing activity against this IFN in a cytopathic effect inhibition assay. Positive samples were assayed in triplicate. Of 391 patients treated for human papillomavirus infections, 13 developed neutralizing antibodies (3.3%); two suffering from condylomata acuminata and 11 from juvenile laryngeal papillomatosis (JLP). None has positive titers prior to receiving IFN-alpha-n1 and none appeared to lose the benefits of IFN treatment. Serial specimens from four JLP patients demonstrated that titers rose during IFN treatment and declined postherapy. None of the 221 cancer patients from whom serum samples were available had positive titers. The overall incidence of IFN neutralizing sera from these IFN-alpha-n1 recipients was 2.1%. PMID- 2553831 TI - Epidemiological observation of coxsackieviruses group B in sewage. AB - Different types of coxsackieviruses group B were isolated from the sewage consecutively annually from January 1982 to March 1987: type 3 in 1982, type 4 in 1983, type 2, 4 and 5 in 1984, type 3 in 1985, and type 4 in 1986 were found. These viruses caused large epidemics among children aged 1 to 4 and proximities. Herald waves for the epidemics were observed six times during this period, and the viruses were detected in a brief period of isolation. With respect to the periodicity of these viruses, the cycles for types 3 and 4 were 2 to 3 years, and longer cycles seemed to be exhibited by types 1, 2, and 5. The onset and termination of the epidemics will only be grasped by surveys throughout the year. HEp-2 cells exhibited the best sensitivity to the viruses. PMID- 2553832 TI - [Criteria of curability in the resection therapy for hepatocellular carcinoma]. AB - The present study was aimed at determining a criteria for curability of hepatic resection in the treatment of hepatocellular carcinomas. The 239 patients hepatectomized from 1973 through 1987 were included for analysis. The patients with the tumor thrombi(V) or intrahepatic metastasis(IM) scattered over the unilateral lobe(IM2) showed very poor prognosis in spite of macroscopically complete resections. Consequently, these conditions were categorized into non curative group(NC). The 5-year survival rate of the tumors less than 2 cm was 42%, which was much better than that of 5 cm or more group but no better than that of 2 to 5 cm group. When the prognosis was compared by an extent of resection among the patients with the tumors of less than 5 cm in size and V(-), the patients with lobectomy showed a 5-year survival rate of 84%, much better than 45% of those with segmentectomy or less range. The surgical margin (SM) negative patients showed a better prognosis than the SM positive patients with a slight difference. The tumor backgrounds of the 39 patients who survived three years with disease free condition were as follows: V(-) in all, no intrahepatic metastasis(IM0) in 79%, no IM2, massive resection for all IM1 in whom IM was confined to the area close to the main tumors, a tumor size of 5 cm or less in 82%, SM(-) in 62%. From the findings described above, we defined a complete resection for V(-) and IM0-1 as a curative condition, of which absolute curative resections(AC) corresponded to the patients with the tumors of 5 cm or less and SM(-), and relative curative ones(RC) to the remainders.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553833 TI - [A case with hepatocellular carcinoma with complete remission due to gamma-IFN therapy]. AB - A 65-year-old male was admitted to our hospital with the aim of closer examination of SOL in cirrhotic liver. By abdominal echography, CT and angiography etc., he was diagnosed as LC with multiple HCC, one in S5 about 2.0 cm, another in S1 about 5.0 cm in diameter, and other four small nodules in right lobe. We treated him with recombinant gamma-IFN at dose of 1.6 X 10(7) units/day for 5 consecutive days biweekly. Although until two months the tumours were gradually enlarged in size, then they became smaller, and at five months later after the beginning of gamma-IFN therapy the tumor stains in angiogram disappeared. Then we performed the surgical treatment. Both macro and microscopical findings of tumor specimens in S5, 2. 2 X 2.0 X 1.9 cm in size, showed complete necrotic tissue. Additionally the yellowish small tumor was found on surface of S8, and it was adenomatous hyperplasia histologically. PMID- 2553834 TI - Experimental endotoxemia in pregnancy: in situ glomerular microthrombus formation associated with impaired glomerular adenosine diphosphatase activity. AB - The mechanism of increased sensitivity for endotoxin in pregnancy as reflected by the formation of microthrombi in renal glomeruli is unknown. It has been shown that reduced glomerular diphosphatase (ADPase) activity in the rat kidney greatly increases the intraglomerular thrombotic tendency. We now studied experimental intraglomerular thrombosis ex vivo in association with glomerular ADPase activity in pregnant and nonpregnant control rats after infusion of either endotoxin or saline solution. Each animal (Wistar rat) was equipped with a permanent vena jugularis catheter and received either endotoxin (1.0 micrograms/kg body weight) (n = 6) or saline solution (n = 5) 7 days before being killed; nonpregnant rats were also treated with endotoxin (n = 5) or saline solution (n = 4). On day 21, before the animals were to be killed, they were anesthetized and their left kidneys were perfused with adenosine diphosphate solution (10 micrograms/ml) and platelet-rich plasma (1 x 10(9) cells/ml). Perfused kidneys were processed for light microscopy, electron microscopy, enzyme cytochemistry at the ultrastructural level, and immunohistology. The results showed decreased ADPase activity exclusively in the glomerular basement membrane of kidneys of pregnant rats treated with endotoxin in contrast to the findings in control rats. In addition, exclusively in the group of endotoxin-treated pregnant rats, significantly increased intraglomerular platelet aggregation could be detected after alternate perfusion ex vivo. We suggest that, in the present model, enhanced susceptibility of glomerular ADPase for endotoxin is due to pregnancy associated factors that have yet to be identified. This increased susceptibility may promote in situ formation of intraglomerular microthrombi. PMID- 2553835 TI - Effect of minoxidil on platelet function and the synthesis of prostaglandins in platelets. AB - At the 12.5 micrograms level, minoxidil prevents the irreversible aggregation of platelets by 2 x 10(-6) mol/L adenosine diphosphate (ADP). Levels of minoxidil greater than 12.5 micrograms cause a reversal of primary aggregation by 2 x 10( 6) mol/L ADP. Aggregation of platelets in response to 125 micrograms of arachidonic acid is measurably reduced by 12.5 micrograms of minoxidil and totally suppressed by 30 micrograms. Concurrent with the inhibition of platelet aggregation, increasing concentrations of minoxidil cause a gradual reduction in the synthesis of prostaglandin E2 (PGE2) and thromboxane B2 (TxB2). In the presence of 100 micrograms of minoxidil, PGE2 is reduced from a control value of 87.7 +/- 2.2 pg/ml to 23.9 +/- 3.2 pg/ml. At this level of minoxidil, TxB2 drops from 105 +/- 3.3 ng/ml to 10.5 +/- 2.6 ng/ml. The effect of minoxidil on platelet aggregation is not associated with increased cyclic adenosine monophosphate synthesis. All data support the conclusion that minoxidil functions (in platelet metabolism) primarily as a cyclooxygenase inhibitor. PMID- 2553836 TI - Protein intake and cation transport in the loop of Henle. AB - High-protein intake enhances maximal urinary concentrating ability and suppresses tubuloglomerular feedback activity in a manner that correlates with enhanced salt reabsorption in the loop of Henle. In this article we describe studies designed to localize the site at which protein intake alters loop sodium uptake (JNa) in rats fed diets containing either 6% or 40% protein for approximately 8 to 10 days. In vivo microperfusion demonstrated that luminal bumetanide (10(-5) mol/L) fully reversed the stimulation of JNa by high-protein intake, thus suggesting that high-protein intake stimulates salt transport in the thick ascending limb. In vitro studies supported this possibility, showing that high-protein intake significantly increased sodium-potassium adenosine triphosphatase (NaK ATPase) activity in homogenates of outer renal medulla (68%) and in dissected medullary thick ascending limbs (87%). This effect was partly selective, since high-protein intake did not alter NaK ATPase activity in superficial renal cortex, had a smaller and statistically insignificant effect on NaK ATPase activity in dissected pars rectae, and did not affect magnesium ATPase activity in any tissue. Furthermore, this effect did not appear to require hypertrophy, since high-protein intake for approximately 8 days did not detectably alter the relative amounts of tissue protein and DNA in either medulla or cortex. A last series of studies demonstrated that high-protein intake increased plasma aldosterone levels. We conclude that increased protein intake stimulates salt reabsorption predominantly in the thick ascending limb, an effect that is partly selective; does not appear to require hypertrophy; and may be related to increased plasma aldosterone levels. PMID- 2553837 TI - Lithium chloride stimulates human monocytes to secrete tumor necrosis factor/cachectin. AB - The purpose of this study was to examine the effect of lithium chloride (LiCl) on human monocytes. Patients undergoing lithium therapy have elevated white blood cell counts. Since both tumor necrosis factor alpha (TNF alpha) and interleukin 1 (IL-1), which are secreted by monocytes, can stimulate endothelial cells to produce granulocyte-macrophage colony-stimulating factor (GM-CSF), we determined whether lithium-stimulated monocytes produced TNF alpha and/or IL-1. Normal human monocytes were incubated for 24 h with medium (negative control), lipopolysaccharide (positive control), or LiCl (0.05-50 mM). The supernatants were removed and assayed for IL-1 and TNF alpha secretion using the D10.G4.01 and L929 assays, respectively. Lithium did not stimulate IL-1 secretion but did stimulate TNF alpha secretion (5-10 U/ml of TNF alpha per 2 x 10(5) monocytes). The increased secretion of TNF alpha was associated with a fourfold increase in TNF alpha mRNA. TNF alpha activity in the supernatants was neutralized by a monoclonal antibody against human TNF alpha but not by antibody against human albumin. Other alkali metals such as rubidium and cesium did not stimulate monocytes to secrete TNF alpha. These data indicate that one mechanism by which Li may cause granulocytosis is through a transcriptional enhancement of TNF production and subsequent secretion by monocytes. PMID- 2553838 TI - Studies on the mechanisms of ACTH-induced inhibition of aldosterone biosynthesis in the rat adrenal cortex. AB - In rats, chronic treatment with high doses of ACTH (10-40 micrograms/100 g body weight per day) leads to a marked reduction in aldosterone synthesis by adrenal capsules. The possibility that this inhibition is secondary to a decrease in plasma potassium levels or in renin angiotensin system (RAS) activity has been explored. The effects of chronic ACTH treatment were compared in (I) animals in which the endogenous RAS activity was stimulated by restricting dietary sodium intake, (II) animals in which plasma angiotensin II was increased by infusion from implanted osmotic minipumps and (III) animals which received supplementary potassium and in which hypokalaemia was prevented. In all cases, rates of aldosterone biosynthesis in vitro by adrenal capsules were decreased in ACTH treated animals to an extent similar to those in untreated controls. In addition, ACTH treatment of hypophysectomized rats resulted in a similar inhibition of aldosterone biosynthesis to that found in sham-operated controls. It may be concluded that the ACTH-induced reduction of aldosterone biosynthesis is independent of the secretion of other pituitary hormones, and cannot be simply ascribed to either a reduction in RAS activity or in plasma potassium levels. The results are consistent with the view that the effects of chronic ACTH treatment are mediated by a direct action on the zona glomerulosa cell, which leads to its transformation into a zona fasciculata-like form. PMID- 2553839 TI - Proliferation and differentiation of possible Leydig cell precursors after destruction of the existing Leydig cells with ethane dimethyl sulphonate: the role of LH/human chorionic gonadotrophin. AB - The influence of LH levels on the proliferation and differentiation of possible Leydig cell precursors was investigated in adult rats, after the destruction of the existing Leydig cells with the cytotoxic drug ethane dimethyl sulphonate (EDS). In rats bearing a testosterone implant which prevented the rise in plasma LH levels and kept them within the normal range after the destruction of the Leydig cells, the proliferative activity of possible Leydig cell precursors still increased seven- to eightfold 2 days after EDS administration. Apparently, in this situation, locally produced factors, and not LH, may play a role in the stimulation of proliferation. The proliferative activity of the possible precursor cells could be further stimulated by treating rats with daily injections of human chorionic gonadotrophin (hCG) following EDS administration. It was concluded that the proliferative activity of possible Leydig cell precursors is probably regulated by both paracrine and endocrine factors. Almost no Leydig cells were formed in the rats bearing a testosterone implant during the first 4 weeks after EDS administration. When these rats were treated with hCG, starting 28 days after administration of EDS, a substantial number of Leydig cells was found after 2 days, and these cells also showed 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) and alpha-naphtyl esterase (alpha-NE) activity. When hCG treatment was started at 14 or 21 days after EDS administration, some cells with the nuclear characteristics of Leydig cells were present after 2 days, but no 3 beta-HSD or alpha-NE activity could be detected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553840 TI - Quantification of opioid-binding sites in the ewe hypothalamus. AB - Opioid-binding sites were quantified in the ewe hypothalamus using [3H]diprenorphine ([3H]DIP) as the radioligand. [3H]DIP binding to hypothalamic membrane preparations was stereospecific, saturable with respect to [3H]DIP concentration, and linear with hypothalamic membrane protein content. Scatchard analysis revealed a single class of binding sites. There were no significant differences in binding site concentration or binding affinity in hypothalami from intact ewes during the breeding and non-breeding seasons, or from long-term ovariectomized ewes with and without oestradiol treatment during the breeding season. Thus, whilst ovarian steroid hormones are known to modify LH responses to opioids and their antagonists in the ewe in vivo, they do not appear to do this by modulating the numbers of hypothalamic opioid-binding sites. PMID- 2553841 TI - Characterization of endometrial and myometrial oxytocin receptors in the non pregnant ewe. AB - Oxytocin-binding sites in the endometrium and myometrium of the non-pregnant ewe were characterized. [3H]Oxytocin bound to a single site in both tissues with high affinity; dissociation constants were determined to be 1.96 nmol/l in endometrium and 2.12 nmol/l in myometrium. Oxytocin binding was enhanced by divalent cations with a similar order of potency in both tissues: Co2+ greater than Mn2+ greater than Ni2+ greater than Mg2+ greater than Zn2+ greater than Ca2+. The endometrial and myometrial binding sites showed the same specificity for oxytocin analogues and related peptides, having high affinity for oxytocin, [Arg8]-vasopressin, [Lys8]-vasopressin, and the oxytocin-specific agonists [Gly7]-oxytocin and [Thr4,Gly7]-oxytocin. The results suggest that oxytocin receptors present in the endometrium and myometrium of the ewe are similar both to each other and to classical oxytocin receptors. PMID- 2553842 TI - Changes in serum concentrations of ACTH, corticosterone and in growth following morning or evening injection of female rats with porcine growth hormone. AB - The present study was designed to examine the effects of stress, associated with daily handling and placebo injection, on growth and adrenal activity in female rats. A second objective was to examine the effects of porcine GH (pGH) on growth and on serum concentrations of ACTH and corticosterone. Treatments were administered in the morning (08.00 h) in semi-darkness, or in the evening (20.00 h) towards the end of the light period and were timed to coincide with reported periods of high and low adrenal sensitivity to ACTH. Handling of and s.c. injection of saline into female rats (initial body weight 220 g) each morning for 21 days did not affect growth, food intake or organ weights. In contrast, daily handling and saline injection each evening caused a marked reduction in weight gain (25%), food intake (12%) and liver weight (8%), accompanied by a trend towards an increase in adrenal weight (11%), but no change in serum corticosterone concentrations. In a second experiment, treatment of female rats (initial body weight 180 g) with pGH (5.6 mg/kg) daily at 08.00 h caused a significant improvement in weight gain (17%), but food intake and organ weights were unaltered. Daily injection of rats with pGH at 20.00 h caused a greater relative improvement in weight gain (45%) than did treatment at 08.00 h as the growth impairment caused by the stress of handling was counteracted. The adverse effect of evening injections of saline on food intake was also counteracted by pGH. Changes in liver and adrenal weights, however, were not attenuated by pGH treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553843 TI - Effects of 1,25-dihydroxyvitamin D3 and cortisol on bovine and human parathyroid cells. AB - Incubation of bovine parathyroid cells with 1,25-dihydroxyvitamin D3 (1,25 (OH)2D3) decreased both preproparathyroid mRNA levels and parathyroid hormone (PTH) secretion. There was a fall to 56.6 +/- 13.7% (mean +/- S.E.M.) and 65.1 +/ 9.3% in mRNA levels and PTH secretion respectively at 1 nmol 1,25-(OH)2D3/l, and 41.1 +/- 13.6% and 42.0 +/- 12.1% at 10 nmol 1,25-(OH)2D3/l after 24 h. After 48 h in 0.1 nmol 1,25-(OH)2D3/l, mRNA levels had fallen to 35.3 +/- 12.6% and PTH secretion to 32.1 +/- 5.0%. In human adenomatous cells, however, incubation with 1,25-(OH)2D3 (10 nmol/l) had no effect on either mRNA levels or PTH secretion even after 48 h. This lack of sensitivity of adenomatous cells to 1,25-(OH)2D3 was not due to an absence of receptors (3847 +/- 39 receptors/ng cytosolic protein in adenomatous cells compared with 4068 +/- 371 in bovine cells) or receptors being of low affinity. Cortisol (1 mumol/l) caused a reduction in the number of receptors for 1,25-(OH)2D3 in bovine parathyroid cells of approximately 20% within 24 h of incubation, but no change in affinity. This decrease was accompanied by abolition of the response to 1,25-(OH)2D3 and was reversible, in that withdrawal of cortisol for the final 24 h of incubation was sufficient for the response to return, the number of receptors having returned to control values. These results suggest that only a small percentage of receptors for 1,25 (OH)2D3 in bovine parathyroid cells may be functional at any one time.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553844 TI - Inhibitory effects of amiloride and its analogues on prostaglandin E2-stimulated fluid transport by cultured porcine thyroid cells: evidence for apical membrane Na+ channels. AB - Confluent monolayers of cultured porcine thyroid cells transport fluid from the apical to the basal surface, forming circumscribed zones of detachment (domes) from the culture dish substrate. Fluid transport, as measured by increase in dome height, was stimulated by prostaglandin E2 (PGE2; 1 mumol/l) and inhibited by amiloride (0.1-100 mumol/l). Values of the inhibition constant (Ki) with 95% confidence limits for each of a series of amiloride analogues were: 3',4' dichlorobenzamil (DCB), 0.090 (0.045-0.18) mumol/l; 2',4'-dimethylbenzamil (DMB), 0.14 (0.074-0.27) mumol/l; amiloride, 0.72 (0.33-1.8) mumol/l; 5-(N,N hexamethylene)amiloride (HMA), 17 (5.9-43) mumol/l; 5-(N-ethyl-N isopropyl)amiloride (EIPA), 33 (15-71) mumol/l; and 2-guanidinobenzimidazole, 243 (110-570) mumol/l. Triaminopyrimidine was ineffective at concentrations up to 1 mmol/l. Since DCB and DMB are known to have a higher affinity for Na+/H+ channels, while HMA and EIPA show higher affinity for Na+/H+ antiports, it was concluded that PGE2-stimulated fluid transport involved an apical membrane Na+ channel. PMID- 2553845 TI - High serum erythropoietin levels are normalized during treatment of congestive heart failure with enalapril. AB - Eighteen patients with dilated cardiomyopathy (three female, mean age 57 years), were treated for 48 weeks with enalapril added to digoxin and diuretic therapy for congestive heart failure of New York Heart Association (NYHA) functional class II (three patients). III (eight patients) and IV (seven patients), respectively. Serum levels of erythropoietin (EPO) were raised at the start (37 +/- 12.8 pmol 1(-1); mean +/- SD) and were normalized during enalapril treatment (17.5 +/- 9.9 pmol 1(-1) at 48 weeks; P less than 0.001). Serum EPO correlated at the start with NYHA functional class (r = 0.68; P less than 0.05). Normalization of elevated serum EPO concentrations during treatment with enalapril paralleled clinical and haemodynamic improvement, and probably reflected relief from renal hypoxia. PMID- 2553846 TI - Femoral neuropathy secondary to anticoagulation. AB - On the basis of a case history, the causes, symptoms, investigations, diagnosis and treatment of femoral neuropathy complicating anticoagulation are reviewed. PMID- 2553847 TI - Peripheral neuropathy, coagulopathy and nodular regenerative hyperplasia of the liver in a patient with multiple serologic auto-antibody activities and IgM B cell lymphoma. AB - A patient with IgM-secreting B-cell lymphoma developed demyelinating peripheral neuropathy, nodular regenerative hyperplasia of the liver, coagulopathy, and terminal miliary lymphomatous spreading. Serologic auto-antibody activities directed against myelin associated glycoprotein, prothrombinase complex (i.e. lupus anticoagulant) and smooth muscle cells were detected. It is suggested that there is a link between the IgM gammopathy, the immunological abnormalities and the various clinico-pathological conditions. PMID- 2553848 TI - Analysis of peptide binding patterns in different major histocompatibility complex/T cell receptor complexes using pigeon cytochrome c-specific T cell hybridomas. Evidence that a single peptide binds major histocompatibility complex in different conformations. AB - The interaction of TCR, antigen, and MHC complex has been analyzed using synthetic peptide antigens and a series of single amino acid-substituted analogues. Two similar antigens, mouse cytochrome c (mcyt c) and pigeon cytochrome c (pcyt c), elicit T cell responses in strains of mice bearing MHC class II Ek beta Ek alpha (B10.A), Eb beta Ek alpha [B10.A(5R)], and Es beta Ek alpha [B10.S(9R)]. The immunogenic regions of these antigens are located in the peptide sequence p88-104 for pcyt c and m88-103 for mcyt c. The limited T cell repertoire for these antigens is comprised of four groups of T cell phenotypes that have very few differences in their TCR gene make up. In this paper, we examine the diversity in their fine specificity for each of the antigens, m88-103 and p88-104, complexed with each of the I-Ek haplotypes. Epitopes, i.e., residues that interact with the TCR, and agretopes, i.e., residues in the MHC-binding site, were assigned for the two peptide antigens in the presence of APC bearing E beta kEk alpha, Eb beta Ek alpha, or Eb beta Ek alpha using T cell hybridomas of the phenotypes I, IIIa, and IV. From our results, we conclude that first, the substitution of any residue between 95 and 104 of the cytochrome c peptide changed the antigenic potency of the peptide for at least one of the hybridomas. Second, each T cell type has a different recognition pattern of epitopes and agretopes for a particular antigen-MHC complex, thus, ruling out a static model of T cell recognition, which assigns certain, invariant agretopic residues to the peptide by which it interacts with the MHC molecule independently of the TCR. Third, the same T cell hybridoma responded to the antigens differently when presented on various MHC molecules, implying that overall changes in the MHC groove, as displayed by the three haplotypes, may affect the efficiency in binding the peptide. Fourth, since most of the residues are used as epitopes by at least one of the T cell specificities, the peptide appears to be recognized in a different conformation by each T cell hybridoma phenotype; and, finally, the epitopic and agretopic residues do not segregate, for any one of the T cell specificities, in such a way that suggests they are recognized in a helical conformation. In summary, our results suggest that a single peptide may generate diversity in the T cell response by virtue of its conformational flexibility within the TCR-MHC-antigen complex. PMID- 2553849 TI - Interleukin 6 decreases cell-cell association and increases motility of ductal breast carcinoma cells. AB - Treatment of transformed breast duct epithelial cells with IL-6 produces a unique cellular phenotype characterized by diminished proliferation and increased motility. Human ductal carcinoma cells (T-47D and ZR-75-1 lines) are typically epithelioid in shape and form compact colonies in culture. Time-lapse cinemicrography shows that some untreated cells can transiently become fusiform or stellate in shape and separate from each other within a colony, but they usually rejoin their neighbors. While IL-6 suppresses the proliferation of these carcinoma cells, the IL-6-treated cells generally become stellate or fusiform and show increased motility. These changes persist as long as the cells are exposed to IL-6. This results in the dispersal of cells within colonies. The effects on cell growth, shape, and motility are reversible upon removal of IL-6. IL-6 treated T-47D cells display diminished adherens-type cell junctions, as indicated by markedly decreased vinculin-containing adhesions and intercellular desmosomal attachments. The effects on ZR-75-1 cell shape, colony number, and DNA synthesis are dependent on IL-6 concentration in the range from 0.15 to 15 ng/ml. Higher concentrations are required in T-47D cells for equivalent effects. Anti-IL-6 immune serum blocks IL-6 action. IL-6 represents a well-characterized molecule that regulates both the proliferation and junction-forming ability of breast ductal carcinoma cells. PMID- 2553850 TI - The association between dietary intake and reported history of Candida vulvovaginitis. AB - The association between dietary intake and the history of Candida vulvovaginitis was evaluated in 166 women who had a history of Candida vulvovaginitis in the past 5 years (cases) and in 207 women without such a history (total population), as well as in 74 women with five or more episodes in the past 5 years and 125 women with no history of Candida vulvovaginitis. Women were interviewed about their demographic data, past medical and sexual history, and their history of vaginal or pelvic infections. An extensive dietary history was taken to determine each woman's usual adult dietary intake. Results indicate associations between total caloric intake, carbohydrates, and fiber and a history of Candida vulvovaginitis. The results were not altered by controlling for age, body mass index, smoking, use of oral contraceptives, and sexual activity variables. These results suggest several dietary constituents may influence susceptibility to Candida vulvovaginitis infections. A follow-up prospective study, using culture confirmation of Candida infection, is needed. PMID- 2553852 TI - Acquisition of endogenous ecotropic MuLV can occur before the late one-cell stage in the genital tract of SWR/J-RF/J hybrid females. AB - Endogenous ecotropic MuLV proviral loci are acquired by the progeny of some [SWR/J x (SWR/J x RJ/J)F1] N2 hybrid females obtained by two successive backcrosses of RF/J mice onto the SWR/J background. This results most likely from an infection of early embryos or oocytes by MuLV particles originating from maternal tissues. However, the time and site of infection are not yet known. Using oviductal transfers of embryos at the one-cell stage, we show here that three of 88 N3 embryos from [SWR/J x (SWR/J x RF/J)F1] N2 hybrid females transferred to virus-free foster mothers harbored new proviral integrations, whereas none of 61 SWR/J embryos transferred to [SWR/J x (SWR/J x RF/J)F1] N2 hybrid females had acquired any proviruses. These data support the infection of oocyte and/or early one-cell embryo as the initial event leading to new proviral insertions. PMID- 2553851 TI - Dolichol phosphorylation occurs via a CTP-dependent reaction in Artemia larvae. AB - The phosphorylation of dolichol in larval stages of the brine shrimp, Artemia salina, has been investigated. The dolichol kinase has been assayed in crude microsomes; the enzyme requires CTP as phosphoryl donor and calcium as divalent cation. Activity increases with both incubation time and added microsomal protein. The product of the reaction has been characterized by chromatographic and enzymatic procedures. With gamma-32P CTP as substrate, the apparent Km for CTP is 24 microM. Enzymatic activity is stimulated fivefold by exogenous dolichol. The specific activity of the enzyme increases with the frequency of molting. Dolichol kinase activity was detectable in membranes prepared from dormant Artemia cysts. The low level in dormancy may anticipate the critical role of the enzyme during hatching. PMID- 2553853 TI - Genetic analysis of erythrocyte uridine monophosphate kinase and aminolevulinate dehydrase and its application to paternity testing. AB - Simultaneous determination of uridine monophosphate kinase (UMPK) and aminolevulinate dehydrase (ALAD) has been carried out after discontinuous starch gel electrophoresis in the Galician population (NW Spain), including 129 families with a total of 291 descendents. Formal genetic studies are in agreement with the autosomal codominant way of inheritance for each locus. No evidence of phenotype association between both loci among the offspring is observed. Chance of exclusion for non-fathers is 0.041 13 for UMPK and 0.0702 for ALAD configuring a total exclusion rate of 0.1085 when both systems are evaluated together. PMID- 2553854 TI - Multiple-hormone-producing islet cell carcinoma: report of a case. AB - Although part of pancreatic endocrine tumors may be multihormonal by immunohistochemical study, the clinical manifestations are often related to hypersecretion of only one type of peptide hormone. Only a few cases have shown two or more syndromes simultaneously or transition of one type of syndrome to another with the passage of time. The case reported here is an islet cell carcinoma with liver metastases. The clinical manifestations changed from Zollinger-Ellison syndrome to hypoglycemic syndrome with the secretion of gastrin, insulin and serotonin. The immunohistochemical study confirmed multihormonal production. The elevation of growth hormone with acromegaly was also noted in this case. It remains to be seen if the excessive production of growth hormone was due to growth hormone-releasing hormone secreted by the endocrine pancreatic tumor or to the possible presence of a pituitary microadenoma as a component of the multiple endocrine neoplasia type I syndrome. PMID- 2553855 TI - Interactions of protons with single open L-type calcium channels. pH dependence of proton-induced current fluctuations with Cs+, K+, and Na+ as permeant ions. AB - We studied the pH dependence of the proton-induced current fluctuations that appear in single open L-type Ca channels when monovalent ions are the charge carriers. We used different methods of analysis to obtain kinetic measurements even under conditions where the individual transitions were too fast to be resolved directly as discrete current steps between two conductance levels. The reciprocal of the dwell times at the high conductance level increased linearly with the pipette proton activity, with a slope that was similar for Cs, K, and Na as permeant ions. Contrary to the expectation for a simple model in which the high and low conductances represent the unprotonated and protonated states of the channel, respectively, the dwell times at the low conductance level were also pH dependent and lengthened with increasing proton activity. At all pH values the dwell times at the low conductance level were longest with Cs as permeant ion and shortened in the order Cs greater than K greater than Na. We introduce a more general model of the protonation cycle in which the channel is represented by four states and can be protonated and deprotonated both at the high and low conductance levels. The conductance change is represented by a conformational change of the channel protein. We discuss the validity of this model and its implications for the mechanism by which protons interact with ion permeation through L-type Ca channels. PMID- 2553856 TI - Apical electrogenic NaHCO3 cotransport. A mechanism for HCO3 absorption across the retinal pigment epithelium. AB - Intracellular microelectrode techniques and intracellular pH (pHi) measurements using the fluorescent dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) were employed to characterize an electrogenic bicarbonate transport mechanism at the apical membrane of the frog retinal pigment epithelium (RPE). Reductions in apical concentrations of both [HCO3]o (at constant Pco2 or pHo) or [Na]o caused rapid depolarization of the apical membrane potential (Vap). Both of these voltage responses were inhibited when the concentration of the other ion was reduced or when 1 mM diisothiocyano-2-2 disulfonic acid stilbene (DIDS) was present in the apical bath. Reductions in apical [HCO3]o or [Na]o also produced a rapid acidification of the cell interior that was inhibited by apical DIDS. Elevating pHi at constant Pco2 (and consequently [HCO3]i) by the addition of apical NH4 (20 mM) produced an immediate depolarization of Vap. This response was much smaller when either apical [HCO3]o or [Na]o was reduced or when DIDS was added apically. These results strongly suggest the presence of an electrogenic NaHCO3 cotransporter at the apical membrane. Apical DIDS rapidly depolarized Vap by 2-3 mV and decreased pHi (and [HCO3]i), indicating that the transporter moves NaHCO3 and net negative charge into the cell. The voltage dependence of the transporter was assessed by altering Vap with transepithelial current and then measuring the DIDS-induced change in Vap. Depolarization of Vap increased the magnitude of the DIDS-induced depolarization, whereas hyperpolarization decreased it. Hyperpolarizing Vap beyond -114 mV caused the DIDS-induced voltage change to reverse direction. Based on this reversal potential, we calculate that the stoichiometry of the transporter is 1.6-2.4 (HCO3/Na). PMID- 2553857 TI - Calcium currents in bullfrog sympathetic neurons. II. Inactivation. AB - Calcium currents in bullfrog sympathetic neurons inactivate slowly and partially during depolarizations lasting 0.5-1 s. There is also a slower (minutes) inactivation process with a broad voltage dependence. An irreversible loss of current (rundown) is prominent with low concentrations of intracellular Ca2+ buffers, with either Ca2+ or Ba2+ as the charge carrier. The extent and rate of the more rapid inactivation process are maximal near the voltage at which the peak inward current is generated, suggesting that inactivation might be Ca2+ dependent. However, inactivation occurs with either Ca2+ or Ba2+ as the charge carrier, is not prevented by strong buffering of intracellular Ca2+ with 10 mM BAPTA, and varies little as the peak current is changed 10-fold by changing the divalent ion concentration. That is, rapid inactivation is not explained by simple versions of voltage, Ca2+- or current-dependent inactivation models. A model in which ion binding within the channel allows a slower, rate-limiting inactivation process fits some but not all of the observed features of inactivation. A purely voltage-dependent three-state cyclic model fits the data if microscopic inactivation is favored by hyperpolarization. PMID- 2553860 TI - Different regulation of serotonin receptors following adrenal hormone imbalance in the rat hippocampus and hypothalamus. AB - Adrenal influence on serotonin (5-HT) transmission in the hippocampal and hypothalamic areas was studied in adrenalectomized rats receiving or not corticosterone replacement. After adrenalectomy, the 5-HT presynaptic receptors were desensitized both in hippocampus and hypothalamus: a significant increase in 5-HT 1 and 5-HT 2 receptor binding numbers took place in membranes from the hippocampus, but not in hypothalamus, while no changes in affinity of receptors to radioligands were observed in either brain area. Corticosterone treatment restored the adrenalectomy-impaired 5-HT autoreceptor sensitivity in hippocampus and hypothalamus and 5-HT density receptor sites in the hippocampus. Serotonin autoreceptor down-regulation following adrenalectomy may increase 5-HT release to maintain the constancy of serotonergic transmission in the brain and 5-HT modulated CRH-ACTH release to compensate the plasma corticosteroid drop. Corticosterone seems to display a distinct tonic control on serotonin transmission in both hippocampus and hypothalamus, the diversity being due to the different roles played by the hormone in these brain regions. PMID- 2553858 TI - Interactions of protons with single open L-type calcium channels. Location of protonation site and dependence of proton-induced current fluctuations on concentration and species of permeant ion. AB - We further investigated the rapid fluctuations between two different conductance levels promoted by protons when monovalent ions carry current through single L type Ca channels. We tested for voltage dependence of the proton-induced current fluctuations and for accessibility of the protonation site from both sides of the membrane patch. The results strongly suggest an extracellular location of the protonation site. We also studied the dependence of the kinetics of the fluctuations and of the two conductance levels on the concentration of permeant ion and on external ionic strength. We find that saturation curves of channel conductance vs. [K] are similar for the two conductance levels. This provides evidence that protonation does not appreciably change the surface potential near the entry of the permeation pathway. The proton-induced conduction change must therefore result from an indirect interaction between the protonation site and the ion-conducting pathway. Concentration of permeant ion and ionic strength also affect the kinetics of the current fluctuations, in a manner consistent with our previous hypothesis that channel occupancy destabilizes the low conductance channel conformation. We show that the absence of measurable fluctuations with Li and Ba as charge carriers can be explained by significantly higher affinities of these ions for permeation sites. Low concentrations of Li reduce the Na conductance and abbreviate the lifetimes of the low conductance level seen in the presence of Na. We use whole-cell recordings to extrapolate our findings to the physiological conditions of Ca channel permeation and conclude that in the presence of 1.8 mM Ca no proton-induced fluctuations occur between pH 7.5 and 6.5. Finally, we propose a possible physical interpretation of the formal model of the protonation cycle introduced in the companion paper. PMID- 2553861 TI - Blockage of nerve growth factor action in PC12h cells by staurosporine, a potent protein kinase inhibitor. AB - Staurosporine, which has a structure similar to that of K-252a, a potent protein kinase inhibitor that blocks nerve growth factor (NGF) action in PC12 and PC12h cells, is also known as a potent inhibitor of several protein kinases. This study shows that in PC12h cells staurosporine has a dual action: at lower concentrations than that required by K-252a, it is an inhibitor of NGF induction of neurite formation and of changes in the phosphorylation of specific proteins, whereas at concentrations higher than that required to inhibit NGF-induced neurite outgrowth, it rapidly enhances outgrowth by itself. PMID- 2553859 TI - Nonlinear charge movement in mammalian cardiac ventricular cells. Components from Na and Ca channel gating. AB - Intramembrane charge movement was recorded in rat and rabbit ventricular cells using the whole-cell voltage clamp technique. Na and K currents were eliminated by using tetraethylammonium as the main cation internally and externally, and Ca channel current was blocked by Cd and La. With steps in the range of -110 to -150 used to define linear capacitance, extra charge moves during steps positive to approximately -70 mV. With holding potentials near -100 mV, the extra charge moving outward on depolarization (ON charge) is roughly equal to the extra charge moving inward on repolarization (OFF charge) after 50-100 ms. Both ON and OFF charge saturate above approximately +20 mV; saturating charge movement is approximately 1,100 fC (approximately 11 nC/muF of linear capacitance). When the holding potential is depolarized to -50 mV, ON charge is reduced by approximately 40%, with little change in OFF charge. The reduction of ON charge by holding potential in this range matches inactivation of Na current measured in the same cells, suggesting that this component might arise from Na channel gating. The ON charge remaining at a holding potential of -50 mV has properties expected of Ca channel gating current: it is greatly reduced by application of 10 muM D600 when accompanied by long depolarizations and it is reduced at more positive holding potentials with a voltage dependence similar to that of Ca channel inactivation. However, the D600-sensitive charge movement is much larger than the Ca channel gating current that would be expected if the movement of channel gating charge were always accompanied by complete opening of the channel. PMID- 2553862 TI - Repetitive action potentials in isolated nerve terminals in the presence of 4 aminopyridine: effects on cytosolic free Ca2+ and glutamate release. AB - The mechanisms by which an elevated KCl level and the K+-channel inhibitor 4 aminopyridine induce release of transmitter glutamate from guinea-pig cerebral cortical synaptosomes are contrasted. KCl at 30 mM caused an initial spike in the cytosolic free Ca2+ concentration ([Ca2+]c), followed by a partial recovery to a plateau 112 +/- 13 nM above the polarized control. The Ca2+-dependent release of endogenous glutamate, determined by continuous fluorimetry, was largely complete by 3 min, by which time 1.70 +/- 0.35 nmol/mg was released. [Ca2+]c elevation and glutamate release were both insensitive to tetrodotoxin. KCl-induced elevation in [Ca2+]c could be observed in both low-Na+ medium and in the presence of low concentrations of veratridine. 4-Aminopyridine at 1 mM increased [Ca2+]c by 143 +/- 18 nM to a plateau similar to that following 30 mM KCl. The initial rate of increase in [Ca2+]c following 4-aminopyridine administration was slower than that following 30 mM KCl, and a transient spike was less apparent. Consistent with this, the 4-aminopyridine-induced net uptake of 45Ca2+ is much lower than that following an elevated KCl level. 4-Aminopyridine induced the Ca2+-dependent release of glutamate, although with somewhat slower kinetics than that for KCl. The measured release was 0.81 nmol of glutamate/mg in the first 3 min of 4 aminopyridine action. In contrast to KCl, glutamate release and the increase in [Ca2+]c with 4-aminopyridine were almost entirely blocked by tetrodotoxin, a result indicating repetitive firing of Na+ channels. Basal [Ca2+]c and glutamate release from polarized synaptosomes were also significantly lowered by tetrodotoxin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553863 TI - Increased uric acid in the developing brain and spinal cord following cytomegalovirus infection. AB - Tissue concentrations of uric acid were determined in the spinal cord, cerebellum, caudate-putamen, and cerebral cortex of developing mice following intraventricular inoculation with murine cytomegalovirus (MCMV) on postnatal day 10. Transient signs of neurological impairment were observed in MCMV-infected animals beginning on days 13-16 and continuing until days 19-21. At the onset of neurological impairment, uric acid concentrations in tissues from infected animals were 17-60-fold greater than in control animals. On postnatal day 70, 60 days after inoculation and 40 days after resolution of neurological signs, uric acid levels were still two- to threefold greater in infected animals. Histological examination revealed signs of focal ischemia in the cerebral and cerebellar cortices of MCMV-infected mice only at the onset of neurological impairment, with ischemic cell changes in some pyramidal neurons of the cerebral cortex. These results indicate that uric acid may be a sensitive marker of persistent vascular pathology resulting from cytomegalovirus infection of the developing nervous system. PMID- 2553864 TI - Adrenergic receptors in aging and Alzheimer's disease: increased beta 2-receptors in prefrontal cortex and hippocampus. AB - Loss of pigmented noradrenergic locus ceruleus neurons occurs in Alzheimer's disease (AD) and, to a lesser extent, in aging. We studied beta-adrenergic receptors and their subtypes, beta 1 and beta 2, by the specific binding of 125I pindolol to particulate membrane preparations from prefrontal cortex, hippocampus, putamen, and cerebellum and to sections from frontal cortex by in vitro autoradiography. In prefrontal cortex from controls, numbers of total beta- and beta 2-adrenoceptors did not significantly correlate with age, but number of beta 1-adrenoceptors showed a weak but significant negative correlation. Binding in tissue particulate preparations to total beta-receptors did not reveal significant differences in samples from prefrontal cortex between AD subjects and age-matched controls. However, beta 1-adrenoceptors were decreased and beta 2 adrenoceptors were increased in number by approximately 30-50% in AD subjects. Thus, the relative ratio of beta 1-/beta 2-receptors was decreased in AD. Binding by in vitro receptor autoradiography performed in a subset of samples of frontal cortex also showed beta 2-adrenoceptors, and less consistently total beta- and beta 1-receptors, to be increased significantly in number in cortical laminae II, III, IV, and V of tissue sections from AD subjects. In these subjects, number of locus ceruleus cells and norepinephrine concentrations in putamen and frontal cortex were markedly reduced compared with values in controls. In the hippocampus, total beta- and both beta 2- and beta 1-adrenoceptors were increased in number in AD. In contrast, in the putamen, where beta 1-receptors predominate, total beta- and beta 1-receptors were significantly decreased in number with no consistent change in content of beta 2-receptors in AD. There were no significant changes in the cerebellum. Specific pindolol binding was not affected by interval between death and sampling of tissue at autopsy. Our results indicate selective changes in number of beta-receptors in AD. These changes in the cortex and hippocampus suggest receptor upregulation in response to noradrenergic deafferentation from the locus ceruleus or may simply reflect glial proliferation in AD. PMID- 2553865 TI - Regional variations in alpha 1-adrenergic receptor subtypes in rat brain. AB - alpha 1-Adrenergic receptor subtypes were differentiated by their affinities for the competitive antagonist WB 4101 and their sensitivities to inactivation by chlorethylclonidine (CEC) in eight rat brain regions. WB 4101 showed low Hill coefficients for inhibition of specific 125I-[2-beta-(4 hydroxyphenyl)ethylaminomethyl]tetralone (125IBE) binding in all regions. Nonlinear regression analysis showed that there were two binding sites with different affinities for WB 4101 in each region. The proportions of these sites varied among regions, although the affinity of WB 4101 for each site remained constant. Thalamus and cerebral cortex had the highest proportion of low-affinity sites, whereas hippocampus and pons-medulla had the highest proportion of high affinity sites. Pretreatment with CEC in hypotonic buffer significantly reduced the density of 125IBE binding sites in all brain regions. Cerebral cortex and cerebellum had the highest proportion of CEC-sensitive sites, whereas hippocampus and spinal cord had the highest proportion of CEC-insensitive sites. There was a significant correlation between the proportion of binding sites with a low affinity for WB 4101 and those sensitive to inactivation by CEC. PMID- 2553866 TI - Atrial natriuretic peptide modulates amiloride-sensitive Na+ transport across the blood-brain barrier. AB - We obtained evidence that amiloride specifically potentiates 125I-labeled alpha rat atrial natriuretic peptide (1-28) [atrial natriuretic peptide (ANP)-(99-126); rANP] binding to cerebral capillaries isolated from the rat cerebral cortex. The binding parameters, KD of 173 pM and Bmax of 159 fmol/mg of protein, became 33 pM and 88 fmol/mg of protein, respectively, when 10(-4) M amiloride was added to the incubation medium. When the effect of rANP was investigated on in vitro 22Na+ uptake into isolated cerebral capillaries, 10(-7) M rANP significantly inhibited the uptake in the presence of 1.0 mM ouabain, 1.0 mM furosemide, and 2.0 mM LiCl in the uptake buffer, a finding suggesting a specific inhibitory effect of rANP on amiloride-sensitive Na+ transport. Thus, the possibility that ANPs control amiloride-sensitive Na+ transport at the blood-brain barrier by interacting with specific receptors has to be considered. PMID- 2553867 TI - Does the olfactory system mediate water- and mineral-regulating mechanisms? Evidence of immunoreactive atrial natriuretic factor within olfactory mucosa. AB - The immunoreactivity of atrial natriuretic factor (ANF) was studied in the rat olfactory mucosa (OM). Endogenous immunoreactive ANF (IR-ANF) was purified from OM using Vycor glass beads for extraction and reverse-phase HPLC: two of three IR ANF peaks, identified by retention time, were identical to both the circulating form of ANF (Ser99-Tyr126) and the ANF pro-hormone (Asn1-Tyr126). A radioreceptor assay, employing rat renal glomerular membranes, revealed that endogenous IR-ANF competed with radiolabelled ANF. IR-ANF was localized by immunocytochemistry in secretory cells of Bowman's gland and in some cells of the epithelial layer. The relatively low concentration of IR-ANF in the OM (2.5 ng/mg protein) suggests a local role of ANF in this tissue. This hypothesis is supported by the presence in OM of ANF-binding sites, characterized by a KD of 95 pM and a Bmax of 130 fmol/mg protein. We propose that ANF could be released from the OM and act throughout in a paracrine (if not autocrine) manner on some yet-unidentified targets containing ANF-binding sites. PMID- 2553868 TI - Adenosine formation and release by embryonic chick neurons and glia in cell culture. AB - Adenosine formation and release were studied in 48-h-old cultured ciliary ganglia and confluent peripheral and CNS glial cultures from embryonic chicks. Metabolic poisoning induced by 30 mM 2-deoxyglucose and 2 micrograms/ml oligomycin reduced ATP concentration by 90%. An increase in adenosine accounted for 15-40% of the fall in ATP. Dilazep (3 X 10(-6) M), a nucleoside transport inhibitor, decreased both incorporation of adenosine (an index of nucleoside transport) and release of adenosine by 80-90%. Dilazep trapped the newly formed adenosine intracellularly. A concentration of alpha, beta-methylene ADP that inhibited ecto-5'-nucleotidase by 80-90% did not alter the concentration of adenosine or AMP in neurons, glia, or medium. The results demonstrate that adenosine is formed intracellularly and exported out of the cell via the nucleoside transporter. The participation of ecto-5'-nucleotidase was excluded. PMID- 2553869 TI - Accumulation of cyclic AMP elicited by vasoactive intestinal peptide is potentiated by noradrenaline, histamine, adenosine, baclofen, phorbol esters, and ouabain in mouse cerebral cortical slices: studies on the role of arachidonic acid metabolites and protein kinase C. AB - In mouse cerebral cortical slices, noradrenaline (NA) potentiates cyclic AMP (cAMP) accumulation elicited by vasoactive intestinal peptide (VIP) through alpha 1-adrenergic receptors. This synergism is inhibited by indomethacin, and the prostaglandins E2 and F2 alpha mimic the effect of NA. In the present study, we observed that the synergism between VIP and NA is not inhibited by the protein kinase C inhibitor 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7) or the diacylglycerol-lipase inhibitor RHC 80267, thus further stressing the role of phospholipase A2 activation. Various neuroactive agents that potentiate the stimulatory effect of VIP on cAMP formation were also examined. As with NA, the potentiation by histamine and adenosine is inhibited by indomethacin. In contrast to NA, histamine, and adenosine, the synergistic interaction between phorbol esters and VIP on cAMP formation is abolished by H-7 but not by indomethacin. The potentiation by baclofen, a gamma-aminobutyric acidB receptor agonist, is partially inhibited by the 5-lipoxygenase inhibitor nafazatrom. The synergism between ouabain and VIP is reduced by H-7 but not by indomethacin and nafazatrom. These data indicate that the stimulation of cAMP formation elicited by VIP is under the modulation of various neuroactive agents that trigger diverse intracellular mechanisms to potentiate the effect of the peptide. PMID- 2553870 TI - A kainate receptor linked to nitric oxide synthesis from arginine. AB - In slices of young rat cerebellum, the glutamate analogue kainate induced a large accumulation of cyclic GMP, which was inhibited by non-N-methyl-D-aspartate antagonists. Quisqualate and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate evoked only small cyclic GMP responses and inhibited the effect of kainate. When tested in cerebellar cell suspensions, glutamate was also a potent antagonist of the cyclic GMP response to kainate. Superoxide dismutase enhanced the response in the isolated cells, whereas haemoglobin and methylene blue were inhibitory. The response in slices was Ca2+ dependent, augmented by arginine, and inhibited by L NG-monomethylarginine in a manner that could be reversed by additional arginine. It is concluded that stimulation of kainate receptors leads to activation of the enzyme that synthesizes nitric oxide from arginine and that activation of soluble guanylate cyclase by the released nitric oxide accounts for the cyclic GMP generation. PMID- 2553871 TI - Plant metabolites. Structure and in vitro antiviral activity of quinovic acid glycosides from Uncaria tomentosa and Guettarda platypoda. AB - A reinvestigation of the bark of Uncaria tomentosa afforded, in addition to the major quinovic acid glycosides 1-3, three further glycosides 4-6. The structures were elucidated by spectral and chemical studies. Furthermore, a series of antiviral tests were performed on all these glycosides and on the related glycosides 7-9, previously isolated from Guettarda platypoda. PMID- 2553873 TI - Tissue specificity in cytochrome c oxidase deficient myopathy. AB - Biopsied muscles were treated in 2 ways to demonstrate cytochrome c oxidase (CCO) activity on electron microscopy: (1) one to several muscle fibers were teased off the biopsy in buffer solution after glutaraldehyde fixation, (2) 20-30-microns thick cryostat sections were placed on precooled glass slides and fixed in glutaraldehyde solution at room temperature. After rinsing in buffer, the teased fibers and cryostat sections were stained with cytochrome c oxidase. In both procedures, almost all mitochondria in control muscle fibers stained positively. In CCO deficiency, the enzyme activity differed from tissue to tissue indicating marked tissue specificity. In the fatal infantile form enzyme activity in muscle fibers was absent, but present in fibroblasts, endothelial cells and smooth muscle arterial cells. The enzyme activity in other forms differed from cell to cell, but individual mitochondria in a given cell examined in cross-section showed uniform CCO activity, indicating that there was no intracellular mosaicism of enzyme positive and negative mitochondria. PMID- 2553872 TI - Studies on inhibitors of skin-tumor promotion. Inhibitory effects of triterpenes from Cochlospermum tinctorium on Epstein-Barr virus activation. AB - Arjunolic acid, an oleanene-type triterpene isolated from the rhizome of Cochlospermum tinctorium, its triacetate derivative, and their methyl esters were tested using the short-term in vitro assay on EBV-EA activation in Raji cells induced by 12-O-tetradecanoyl-phorbol-13-acetate (TPA). Their inhibitory effects on skin tumor promotors were found to be greater than those of previously studied natural products. PMID- 2553874 TI - Immunocytochemical profile of neurofibrillary tangles in Down's syndrome patients of different ages. AB - Brains were obtained at autopsy from 24 patients with Down's syndrome, ranging in age from 13 to 71 years. Neurofibrillary tangle containing neurones of the hippocampus were stained using a Palmgren silver method and immunocytochemically (PAP) using antisera to paired helical filament protein, human tau protein and ubiquitin, as primary antibody. Counts of cells stained by each method were compared. In patients under 50 years of age, in whom only a limited number of tangle bearing cells were present, the number of profiles visualized with silver, anti-paired helical filament and anti-tau methods were similar. However, in patients over 50 years of age (and in certain of those under 50), in whom numerous tangles were present, the number of cell profiles visualized with silver and anti-paired helical filament methods were still similar though anti-tau detected fewer positive cells. This was because of the increased presence, in such patients, of extracellular tangles which had "lost" anti-tau immunoreactivity. Such data suggest that although tau protein forms a major antigenic determinant of neurofibrillary tangles in Down's syndrome (as it does in Alzheimer's disease) this protein may only decorate the basic paired helical filament protein skeleton, and is removed by macrophagic activity upon neuronal death. In all patients, anti-ubiquitin revealed fewer tangles than any other method. It is possible that ubiquitin may be present only transiently, within tangles perhaps following initial formation and lasting only as long as the normal protein degradation processes remain viable within the diseased neurone. PMID- 2553876 TI - The extracellular matrix modulates the response of PC12 cells to nerve growth factor: cell aggregation versus neurite outgrowth on 3-dimensional laminin substrata. AB - PC12 cells attach well to plastic culture dishes coated with laminin, collagen, polylysine or a basement membrane extract (2-dimensional substrata) and, in the presence of NGF, extend short neurites within 1-2 days. However, on gels (3 dimensional substrata) reconstituted from a basement membrane extract (RBM), PC12 cells attach extending short processes transiently and within one day, form networks of small aggregates interconnected by process-bearing cells. By 3 days the network collapses into large aggregates that, in media supplemented with NGF, extended a halo of neurites resembling dorsal root ganglia in culture. Time-lapse video recordings indicate that cell motility on RBM gel is accompanied by extensive blebbing as well as extension of processes that attach to and pull together neighbouring cells. These cellular events may contribute to the disruption of the gel underneath aggregates that is apparent when cultures are stained with Coomasie Blue. Ultrastructural studies indicated that aggregates often have zonula adherens-type junctions where cell bodies and processes come in contact. PC12 cells seeded onto gels of laminin alone behave essentially the same as on RBM gels, whereas on collagen gels they behave as on 2-dimensional substrata and extended neurites rather than aggregate. The extent of aggregation increases with greater cell density and is enhanced significantly by NGF. Antisera to NGF reduce the NGF-enhancement of aggregation but do not block aggregation in the absence of NGF. Dibutyryl cAMP or epidermal growth factor, which stimulate process extension and cell division respectively, do not enhance aggregation. However, 3A3, a monoclonal antibody to a laminin/collagen receptor on PC12 cells and antibodies (Fab fragments) to the neural cell adhesion molecule both inhibit cell aggregation. PMID- 2553875 TI - Lymphocytic beta-adrenergic receptors in X-linked muscular dystrophy. AB - Lymphocytic beta-adrenoceptor levels, receptor binding affinity, lymphocytic basal and isoproterenol-stimulated cyclic AMP (cAMP) production and plasma catecholamine levels were studied in 49 patients with neuromuscular diseases and in 10 healthy subjects. Patients with X-linked muscular dystrophy (Duchenne, 13 patients; Becker, 4 patients) showed a significant reduction in lymphocytic beta adrenoceptor densities (35.9 +/- 2.2 fmol/mg protein vs. 49.6 +/- 3.6 fmol/mg, controls; P less than 0.02), whereas the receptor levels for the patients with spinal muscular atrophy (15), polymyositis (10) and for Duchenne carriers (7) did not differ significantly from the corresponding levels for the control subjects. The reduction in the beta-adrenoceptor density was not correlated with the physical disability of the patients. Lymphocytic beta-adrenoceptor affinity (KD) and basal and isoproterenol-stimulated cAMP levels were all comparable to control subjects' values. Catecholamine levels showed mild inconsistent elevations in various patient groups. The results suggest that reduction in cellular beta adrenoceptors is characteristic of X-linked muscular dystrophies. Its relationship to the basic gene defect is unknown. PMID- 2553877 TI - Intrathecal synthesis of virus-specific oligoclonal antibodies in patients with enterovirus infection of the central nervous system. AB - Cerebrospinal fluid (CSF) and serum samples from six patients with enterovirus infections were investigated by isoelectric focusing (IEF) and affinity-mediated immunoblot AMI) for the clonal distribution of entervirus-specific antibodies. In two patients with either acute meningitis or encephalitis and in one patient with a relapse of multiple sclerosis, oligoclonal IgG bands specific for enteroviruses were found predominantly in the CSF, revealing intrathecal synthesis of these antibodies. In three other patients with neurological symptoms probably unrelated to a current enterovirus infection, IEF and AMI disclosed nearly identical patterns of coxsackievirus-B-specific oligoclonal bands in the CSF and serum, indicating diffusion of these antibodies from the serum into the CSF. Although the number of patients in this study is small, the results suggest that intrathecally synthesized enterovirus-specific antibodies may be used as a means of identifying an enterovirus infection of the CNS. PMID- 2553878 TI - Stage IV non-small-cell lung cancer: the guides are perplexed. PMID- 2553880 TI - Clinical characterization of non-small-cell lung cancer tumors showing neuroendocrine differentiation features. AB - In most cases of small-cell lung carcinomas (SCLC) phenotypic features compatible with a neuroendocrine differentiation status can be identified by monoclonal (MOC) antibody-based immunohistological procedures. Similar features can be recognized only in a minority of non-SCLC tumors. During a period of 30 months, all diagnostic non-SCLC biopsies (141 cases) were prospectively analysed for the presence of markers indicative for neuroendocrine differentiation. In 31% of all cases, such a presence could be noticed. Neuroendocrine differentiation (50% to 100% positive-staining tumor cells) was recognized more frequently in adenocarcinoma when compared to large-cell and squamous-cell carcinoma (chi 2 = 9.31, 2 degrees of freedom, P less than 0.01). To investigate whether the clinical behavior of these "neuroendocrine" non-SCLC cases mimics SCLC, a multivariate analysis for prognostic factors was performed. Among other prognostic factors, biopsies containing more than 50% positive-staining tumor cells with the MOC antibody-1 (MOC-1) were recognized as negative prognostic factors. PMID- 2553879 TI - Combination chemotherapy versus single agents followed by combination chemotherapy in stage IV non-small-cell lung cancer: a study of the Eastern Cooperative Oncology Group. AB - During the last decade, the Eastern Cooperative Oncology Group (ECOG) has studied a series of combination chemotherapy regimens in metastatic (stage IV) non-small cell lung cancer (NSCLC). In January 1984, the ECOG activated a randomized study, EST 1583, which concluded the evaluation of combination regimens in phase III trials and initiated the evaluation of single agents exclusively in previously untreated patients. The treatment regimens in EST 1583 consisted of: (1) mitomycin, vinblastine, and cisplatin (MVP); (2) vinblastine and cisplatin (VP); (3) MVP alternating with the regimen cyclophosphamide, doxorubicin, methotrexate, and procarbazine (CAMP); (4) carboplatin followed by the MVP regimen at the time of progression; and (5) iproplatin followed by MVP at the time of progression. From January 1984 to July 1985, 743 patients were entered on this trial and 699 fulfilled the eligibility requirements. The following objective response rates (complete plus partial remissions) were observed: first-line MVP, 20%; VP, 13%; MVP/CAMP, 13%; carboplatin, 9%; iproplatin, 6%; and second-line MVP, 6%. First line MVP produced a significantly higher response rate than the other treatments (P = .03) adjusted for prognostic variables. Using analyses that were adjusted for prognostic covariates, survival for patients treated on a given regimen was compared with survival for all remaining patients. These analyses showed that treatment with carboplatin was associated with longer survival (median survival time, 31.7 weeks; P = .008) while initial treatment with MVP was associated with a trend for shorter survival (median survival time, 22.7 weeks; P = .09). It should be noted that none of these regimens appear to have produced a clinically meaningful prolongation of survival. Similar analyses evaluating time to progression disclosed that carboplatin-treated patients had a significantly longer time to progression (median time to progression, 29 weeks) than all remaining patients (P = .01). Life-threatening and lethal toxicities (toxicity grades 4 and 5) were greater on the combination regimens than on the single agents (P less than .0001). Based on these results, current group-wide ECOG trials in stage IV NSCLC consist of randomized phase II trials evaluating single agents. PMID- 2553881 TI - Anoxia on slow inward currents of immature hippocampal neurons. AB - 1. The effects of brief anoxia (2-4 min) on membrane currents--especially the tetrodotoxin (TTX)-insensitive, Cd2+-sensitive slow inward currents, presumed to be Ca2+ currents--were studied by single-electrode voltage clamp in CA1 and CA3 neurons in submerged hippocampal slices from adult and newborn Wistar rats (PN1 13). 2. In mature neurons, anoxia had no effect on Q-type inward relaxations, but slowly activating C-type outward currents were depressed. The most striking change was the suppression of Ca inward currents (especially the slowly inactivating L-type, by greater than 95%). This effect of anoxia was not sensitive to the N-methyl-D-aspartate (NMDA) receptor blocker, D aminophosphonovalerate. Anoxia also reversibly abolished the NMDA-evoked inward current. 3. In neurons from newborn animals (PN1-6), Q-type inward relaxations and postanoxic outward currents were very small or undetectable. The slow inward (Ca) currents were smaller than in mature cells, but they showed a clearer separation between low-threshold, fast-inactivating and high-threshold, slowly inactivating currents. Both types of current were more resistant to anoxia (mean depression of L-type was by only 53.3 +/- 5.6%, mean +/- SE). 4. In such immature neurons, the NMDA-evoked inward currents were also more resistant to anoxia. 5. By PN7-13, increasing maturation was reflected in 1) larger voltage-dependent inward currents, 2) increasingly evident Q-type relaxations and postanoxic outward currents, and 3) near-complete blockade of inward currents by anoxia (at PN11-13, mean depression of L-type currents was by 98.5 +/- 1.5%). PMID- 2553883 TI - Mathematical calculations in nursing education: whose responsibility is it, anyway? PMID- 2553882 TI - Multifocal glioblastoma with liver metastases in the absence of surgery. Case report. AB - A patient with two intracerebral glioblastomas of differing histology with metastases to the liver in the absence of surgery is reported. The gliomatous nature of the lesions was confirmed by staining for glial fibrillary acidic protein. Histological and immunohistochemical evidence suggests that the metastases arose from the more poorly differentiated of the intracerebral tumors. One of the intracerebral tumors had enhanced expression of the ras p21 oncogene as compared to the other tumors and as compared to nonmalignant brain tissue from this patient. PMID- 2553885 TI - Faculty practice as a requirement for promotion and tenure: receptivity, risk, and threats perceived. AB - The purpose of this study was to determine the receptivity of nurse faculty to the proposal of adding faculty practice as an additional requirement for promotion and tenure and to investigate the perceived risk and threats from introduction of this proposal. Factors that affect the risk faculty perceived are examined. Nurse educators from all faculty ranks in public nursing programs in 15 states in the southern region of the United States served as subjects for this study (N = 280). Findings indicate that nurse faculty are more receptive than resistant to the proposal requiring faculty practice as a requirement for promotion and tenure. A high negative correlation exists between receptivity to the proposal and the risk faculty perceived from implementation of the proposal A measure of the threat to job prerequisites correlated moderately with the risk faculty perceived from the proposal, with specific threats and benefits perceived by faculty identified. Implications for administrators and faculty are discussed. PMID- 2553884 TI - Mathematics anxiety and its effect on drug dose calculation. AB - Eighty learners were randomly selected from 160 first-year nursing students enrolled in an urban community college nursing program in Ontario. They were subsequently divided into control and treatment groups to investigate the effects of different teaching methods on mathematics anxiety and the students' ability to accurately calculate fractional drug doses. The results obtained in this study indicated that there were no statistically significant differences between the control and treatment groups in either mathematics anxiety levels or in arithmetic test performance. These findings counter many of those found in previous investigations. Reasons for these discrepancies are provided along with recommendations for present practice and future research. PMID- 2553886 TI - A comparative study of learning characteristics of RN and generic students. AB - The constructs of locus of control, self-directed learning readiness, and learning style preference are related components of an individual's learning characteristics. If group differences between registered nurse and generic students on these constructs could be documented, then the educational process could be directed toward facilitating educational goal attainment for these two distinct groups of students. Three instruments were used for this study: the Adult Nowicki-Strickland Internal-External Scale (ANS-IE), Guglielmino's Self Directed Learning Readiness Scale (SDLRS), and Rezler's Learning Preference Inventory (LPI). The sample consisted of 175 generic and 170 RN students enrolled in two BSN programs. No significant group differences were found for the three constructs except for the interpersonal mode of learning, a subscale of the LPI. Although no group differences were found, the data showed an ethnic and age effect involving all subjects. Although there are no wide differences in learning characteristics between generic and RN students, the individual variations support the use of teaching strategies that incorporate independent study and self-paced methodologies. PMID- 2553887 TI - Accelerated LPN-RN nursing education: is role socialization lost along the way? AB - In response to a critical shortage of registered nurses, an accelerated transition program for licensed practical nurses (LPNs) was developed and implemented. A study of the program was undertaken to evaluate students' academic achievement and socialization into the RN role. NCLEX-RN scores and the Professional Nursing Questionnaire (PNQ) of Minnick, Yocum and Scherubel were used for evaluation. Experimental program students were compared with students in the second year of conventional two-year nursing programs. Results of role socialization revealed no significant differences between the groups at either the beginning or end of the program. NCLEX-RN scores were not significantly different. The lack of significant results may be attributable to the inability of the PNQ to detect changes over a short time, or to LPNs' knowledge of the RN role prior to program entry. NCLEX score results indicated that comparable academic achievement was possible in the accelerated program. PMID- 2553888 TI - Measuring perceptions of the exemplary nurse. AB - Qualitative and quantitative data identifying and weighting the perceived importance of different attributes of the "exemplary" nurse were collected from nursing respondents at different points in their professional development. Statistically significant differences in the attribute weights reported by fourth year students and graduate nurses were observed. Because their pool was qualitatively different, quantitative comparisons with first-year students were not possible. The results indicate that these methods may be useful in examining the processes whereby the members of different groups (eg, nursing students and faculty) formulate and modify their assumptions about their own and others' professional values. PMID- 2553889 TI - The meaning of health for graduate nursing students. AB - The purpose of this study was to examine the meaning of health for graduate nursing students based on their life experiences. The qualitative descriptive approach of phenomenology was utilized to define the meaning of health for graduate nursing students. The sample consisted of 29 graduate nursing students. From the participants' responses, 68 descriptive expressions were obtained. Based on these expressions and identified elements, a definition of health was proposed that reflected an awareness of a physical as well as a mental state. PMID- 2553890 TI - The role of the nurse educator when the obstetrical nursing student is male. AB - The role of the nurse educator regarding male obstetrical nursing students is to facilitate a more rewarding and successful educational experience for them. The nurse educator can use specific measures to enhance role success and the student's self-concept. The APPA system may be used to accomplish this. The system is a set of strategies that includes avoiding stereotyping, presenting an equitable perspective, providing comparable clinical experiences, and anticipating guidance needed. This will assist the nurse educator in fulfilling role obligations when the obstetrical nursing student is male. PMID- 2553891 TI - Teaching students how to publish in nursing journals: a group approach. AB - A course in writing for publication in every graduate program would create a plethora of innovative projects, replication studies, and reviews of literature for the nursing community. Group participation in the critiquing of manuscripts provides many reviewers and consequently thought-provoking responses for a more clearly written manuscript. The interaction also provides a support system for the writer and allows the student to realize that the manuscript was not only well-written but informative and interesting to others. PMID- 2553892 TI - Strategy for reinforcing drug knowledge and accountability in drug administration. AB - The strategy of including drug-related nursing diagnoses on the nursing care plan can be a helpful tool in fostering drug knowledge and responsibility for nursing care related to drugs. Using the nursing care plan adds meaning to drug cards and other pharmacologic resources. Individualizing drug considerations on the nursing care plan adds another important dimension to nursing practice. Students demonstrated improved proficiency and relevance in applying drug information to client care. PMID- 2553893 TI - Love, beauty, and truth: fundamental nursing values. PMID- 2553894 TI - Community health nursing practice for students in an outreach BSN program in rural North Carolina. PMID- 2553895 TI - A simple apparatus for processing large numbers of specimens for colloidal gold immunoelectron microscopy: application to paired helical filaments of Alzheimer's disease. PMID- 2553896 TI - Differences in response to dilator agents in blood vessels of different types: physiological bases for selectivity. AB - Differences in the activation systems by which contraction is maintained in blood vessels of different types are an important cause of selectivity in the action of dilator agents. The sympathetic nervous system is the main stimulus for contraction in the limb veins whereas intrinsic (or myogenic) tone makes an important contribution to the contraction of resistance vessels. Drugs that inhibit the effect of the sympathetic nervous system are therefore selective dilators of the veins and drugs that prevent membrane depolarization or block calcium channels are strongly selective for resistance vessels. Substances such as the nitrovasodilators that increase intracellular concentrations of cyclic (c)GMP act preferentially on the veins. Those that act by increasing the concentrations of cyclic (c)AMP show little selectivity unless, as with the beta adrenoceptor agonists, they act through receptors whose distribution varies between vessels. Differences in the distribution of receptors may also be important in the action of some other vasoactive substances such as the prostaglandins. PMID- 2553897 TI - Functional aspects of myogenic vascular control. AB - Recent in vivo observations, especially in skeletal muscle, of macro- and microvascular myogenic reactivity and its functional consequences suggest that myogenic regulatory mechanisms contribute directly or indirectly to circulatory homeostasis by exerting the following main functions: (1) A blood pressure induced tonic excitatory function of the vasculature, thereby initiating a pronounced intrinsic myogenic basal tone in the arterial microvessels; (2) a homeostatic resistance function (high total peripheral resistance), mainly exerted by the myogenic tone and serving to maintain normal arterial pressure at rest; (3) functions serving to improve nutritional flow and exchange, exemplified by blood flow recruitment, capillary recruitment, adjustments to the capillary perfusion:diffusion ratio for optimum exchange in the heterogeneous capillary network, and reactive hyperaemia; and (4) protective functions directed against harmful circulatory effects of changes in blood pressure, exemplified by autoregulation of blood flow and autoregulation of capillary hydrostatic pressure during changes in arterial pressure, and by autoregulation of transcapillary filtration during a hydrostatic load on the vascular bed. PMID- 2553898 TI - Receptor-mediated C-kinase activation contributes to alpha-adrenergic tone in rat mesenteric resistance artery. AB - Small branches from the superior mesenteric arteries (100-200 microns outer diameter) freshly dissected from male Wistar-Kyoto (WKY) rats were mounted for tension recordings. Some arterial rings were left in physiological salt solution and used as intact arteries while others were made permeable with alpha-toxin and incubated in cytoplasmic substitution solution. The relationship between ambient [Ca2+] and tension development during various modes of activation was measured in both intact and permeable arterial rings. The effects of ryanodine and 12-O tetradecanoyl phorbol-13-acetate (TPA) were tested. Ryanodine had no effect on the tone developed in response to noradrenaline, but tension was increased when the tissues were bathed in 80 mmol/l K+ and [Ca2+] was raised (10 mmol/l). If it is assumed that ryanodine acts exclusively to enhance the permeability of the sarcoplasmic reticulum in smooth muscle, these data suggest that noradrenaline induced tone is partly due to inhibition of Ca2+ buffering in the sarcoplasmic reticulum. However, this action of noradrenaline is not as pronounced in the resistance arteries as it is in the rabbit aorta. 12-O-tetradecanoyl phorbol-13 acetate had no effect on the noradrenaline-induced contractions of the intact resistance arteries, but caused a large leftward shift in the relationship between tension and extracellular [Ca2+] when high-K+ depolarization was the stimulus. This increased sensitivity to extracellular [Ca2+] could not be explained by stimulation of the Ca2+ influx. Instead, application of TPA to the rings made permeable with alpha-toxin dramatically increased the myofilament sensitivity to Ca2+, as demonstrated by a shift to the left of the tension intracellular-[Ca2+] curve.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553899 TI - Comparative studies of tissue inhibition by angiotensin converting enzyme inhibitors. AB - There is increasing evidence that inhibition of tissue angiotensin converting enzyme (ACE) is important for the pharmacokinetics and pharmacodynamic effects of ACE inhibitors. Radioligand inhibitor binding methods using 125I-351A and either tissue homogenates or in vitro autoradiography have allowed in vitro and ex vivo quantitation of tissue ACE inhibition by a variety of ACE inhibitors. The rank order of potency against plasma as well as lung, kidney, and cardiac homogenates was quinaprilat = benazeprilat greater than perindoprilat greater than lisinopril greater than enalaprilat greater than fosinoprilat. The highest concentration of ACE in the heart was found in the cardiac valves followed by the right and left atria, then the right and left ventricles. Ex vivo studies showed that after oral administration of quinapril, ACE was inhibited dose-dependently in the lung, kidney, aorta and heart for more than 24h. Tissue bioavailability of the inhibitor is also an important determinant of tissue ACE inhibition. Perindopril crossed the blood-brain barrier and inhibited brain ACE at high doses, but after equivalent doses of quinapril no brain ACE inhibition could be demonstrated. These results suggest that it may be possible to design ACE inhibitors to have specific effects on ACE in different tissues. PMID- 2553900 TI - Does the duration of action of angiotensin converting enzyme inhibitors affect their safety and adverse effects? AB - The results of more than 1600 patient years of experience with the new angiotensin converting enzyme (ACE) inhibitor, quinapril, suggest that it is safe for the treatment of hypertension and congestive heart failure. A once-daily regimen of quinapril minimizes adverse effects on renal function compared with a twice-daily regimen, and compared with enalapril. This may be because long-acting agents, or frequent doses of ACE inhibitors, produce a prolonged reduction in the glomerular filtration pressure. PMID- 2553901 TI - Bilateral parotid gland metastasis as the initial presentation of a small cell lung carcinoma. PMID- 2553902 TI - Antral grafting for endosseous implants. PMID- 2553903 TI - [An experimental study of herpes simplex virus infection in the facial nerve]. AB - In the experimental animals, the author succeeded in establishing the pathogenicity of herpes simplex virus (HSV) in the facial nerve. KOS strain of HSV type 1 was inoculated to the tongue or the auricle of the balb/c mouse previously treated by intraperitoneal injection of cyclophosphamide. Between 4 and 12 days after the inoculation, the facial nerves were resected and submitted to histopathological and immunohistochemical investigations. Inflammatory changes were mainly observed in the geniculate ganglion and its proximal portion of the facial nerve irrespective of the site of the inoculation. In the same sites, HSV antigens were proved. The viral geniculate ganglionitis was induced in 21.4% of the animals inoculated to the tongue, 30.0% of those to the unilateral auricle, and 37.5% of those to the bilateral auricles respectively. This experiment demonstrated that HSV inoculated in the site innervated by sensory component of the facial nerve could affect the geniculate ganglion and its neighboring portion. These results presented a substantial support of the viral hypothesis in the etiology of Bell's palsy. PMID- 2553904 TI - Altered synaptic transmission in Drosophila hyperkinetic mutants. AB - Synaptic transmission in Drosophila can be altered by mutations in specific genes. For example, mutations in the Shaker (Sh) gene, which encodes the rapidly inactivating A-type potassium channel, cause repetitive nerve firing and prolonged transmitter release at the neuromuscular junction. Here we show that mutations in the Hyperkinetic (Hk) gene also affect the properties of synaptic transmission at the neuromuscular junction. In particular, we find that whereas single or low frequency nerve stimulation evokes a wild type postsynaptic response, at higher frequencies of nerve stimulation, each stimulus results in repetitive nerve firing and increased postsynaptic response, which is similar to that observed in Sh mutants. Various experiments suggest that this increased postsynaptic response results from prolonged depolarization of the nerve terminal, leading to increased transmitter release at the neuromuscular junction. The similarity in phenotypes between Sh and Hk mutants, along with the observation that Sh is epistatic to Hk in its effects on synaptic transmission, suggest that Hk acts on synaptic transmission by an effect on A-type potassium channels. PMID- 2553905 TI - Osteoclasts are present in the giant cell variant of malignant fibrous histiocytoma. AB - We assessed the osteoclast-like giant cells in a giant cell-rich variant of malignant fibrous histiocytoma (MFH) for characteristics which are specific for osteoclasts, including excavation of bone and binding of osteoclast-specific monoclonal antibodies. Excavations characteristic of osteoclastic activity were found when the bone slices were inspected in the scanning electron microscope after incubation. Two monoclonal antibodies which bind specifically to osteoclasts showed strong membrane reactivity. These results provide strong evidence for the osteoclastic nature of the giant cells in this variant of the MFH. Osteoblasts are known to control the function of the osteoclast, and since there is bone formation present in at least half of the giant cell-rich variant of MFH it may be that it is the cells with properties peculiar to osteoblasts within the tumour which are responsible for the recruitment of the osteoclasts. PMID- 2553906 TI - An observation of DNA ploidy, histological grade, and immunoreactivity for tumour related antigens in primary and metastatic breast carcinoma. AB - A cohort of 36 cases of advanced breast carcinoma were included in the study. DNA ploidy, histological grade, and immunoreactivity for carcino-embryonic antigen (CEA), carcino-embryonic antigen with normal cross-reacting antigen (CEA/NCA), and to NCRC-11 were assessed in primary breast carcinomas and metastases from the ovary and axillary lymph nodes. Changes in these variables between primary tumours and their metastases were found in 44 per cent of cases for DNA ploidy, 44 per cent of cases for histological grade, 17 per cent of cases for immunoreactivity to NCRC-11, 28 per cent of cases for CEA, and 56 per cent of cases for CEA/NCA immunoreactivity, respectively. Alteration of one factor appeared to be independent of changes in other factors. The development of clones with metastatic potential does not appear to be related to consistent changes in any one of the factors examined and suggests that none of these factors would be of prognostic value in advanced breast carcinoma. PMID- 2553907 TI - Haematoporphyrin derivative (Photofrin II) photosensitization of isolated mitochondria: inhibition of ADP/ATP translocator. AB - To gain further insight into the mechanism by which irradiation of mitochondria in the presence of haematoporphyrin derivative (Photofrin II) (PF II) causes impairment of mitochondrial oxidative phosphorylation, the rate of ADP/ATP exchange via the ADP/ATP translocator was measured fluorometrically is isolated rat liver mitochondria. In accord with noncompetitive inhibition, PF II photosensitization decreases the maximum rate of exchange Vmax (20.8 and 9.6 nmol ATP effluxed min-1 x mg protein in the control and after 2 min irradiation, respectively) without changing the ADP affinity for the carrier (Km = 5 microM in both cases). Comparison of the rate of oxygen uptake by mitochondria stimulated by either ADP or by the uncoupler carbonyl cyanide 4 (trifluoromethoxy)phenylhydrazone (FCCP) confirms that the adenine nucleotide carrier is a major target of photodynamic action which causes oxidative phosphorylation impairment. PMID- 2553908 TI - Liver transplantation in children: the initial Toronto experience. AB - The Hospital for Sick Children's initial 2-year experience with pediatric liver transplantation is reviewed. Patients are divided into high- and low-risk groups according to certain criteria. The high-risk group includes patients under 10 kg in weight, those with extrahepatic biliary atresia (EHBA), those with portal vein atresia or thrombosis, and those in hepatic coma. All others were considered low risk. Twenty-nine patients were assessed for transplantation: 18 were transplanted and 6 (21% of total referred) died while on the waiting list. Eighteen patients received 23 transplants. Of the 18 recipients, nine had EHBA, four had fulminant hepatic failure, two had tyrosinemia, one had glycogen storage disease, one had Indian childhood cirrhosis, and one had idiopathic cirrhosis. Seven of the 13 patients in the high-risk group survived (55% survival) with 1 to 23 month follow-up. Survival was significantly higher (80%) in the low-risk group (P less than 0.05). Four patients were retransplanted and two survived. Early deaths occurred from prolonged warm ischemia, recurrent portal vein thrombosis, and brain death in a patient who had been transplanted in hepatic coma. Late deaths occurred from cytomegalovirus (CMV) disease (2 patients), acute rejection (1 patient), and myocardial infarction (1 patient). The incidence of primary nonfunction was 4.3% (1 of 23) and of arterial thrombosis was 13% (3 of 23). Survival in patients transplanted for EHBA (67%) was slightly higher than it was for the rest of the group, although not as good as it was in the low-risk group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553909 TI - Malignant small-cell tumor of the thoracopulmonary region: 'Askin tumor'. AB - Malignant small-cell tumor of the thoracopulmonary region (MSCT) is an uncommon neoplasm in children. We describe five cases diagnosed since 1981 that fulfill the criteria put forth by Askin et al. Surgery was performed for diagnosis or therapy in all patients. Two patients underwent open lung biopsy, only because of tumor extent. The other three had chest wall resections. All patients received radiotherapy and chemotherapy. Three patients presenting initially with extensive disease died at intervals of 2.5 to 7 months after diagnosis. Two patients are alive and disease-free at 16 and 24 months postdiagnosis. All five cases were reviewed for standard histology and differential immunohistochemistry. Electronmicroscopy and tissue cultures were done in 3 of the 5 patients. All five patients were neuron-specific enolase-positive. MSCT is an exclusion diagnosis not always readily made. Clinical course and pathologic expertise may point to the correct diagnosis. PMID- 2553910 TI - Left hepatic trisegmentectomy for interlobar hepatoblastoma located close to the hepatic hilum. AB - A 10-month-old boy was noted to have hepatomegaly, and hepatic angiography showed a huge tumor of the medial segment of the left lobe, located close to the hepatic hilum and involving the anterior segment of the right lobe in contiguity. We considered at first that resection would not be possible, but at operation the anterior segmental branch of the right hepatic artery was successfully ligated and divided, which made left trisegmentectomy possible in this case. Postoperatively, the patient was treated with cisplatin and tetrahydropyranyl Adriamycin he was doing well without any evidence of disease 24 months after the hepatectomy. PMID- 2553911 TI - Hepatoblastoma in siblings. AB - Two infants, a brother and sister, with hepatoblastoma of the right lobe of the liver were treated and are alive and lead normal lives. Only three other familial occurrences of this tumor have so far been documented. PMID- 2553912 TI - Turcot's syndrome: a diagnostic consideration in a child with primary adenocarcinoma of the colon. AB - We report the diagnosis of an adenocarcinoma of the colon in a 12-year-old girl in association with the presence of a small number of adenomatous polyps and a positive family history of a sibling with a central nervous system glioma. These findings implicate Turcot's syndrome as the cause for the development of intestinal and intracranial neoplasms in the two siblings. Since primary adenocarcinoma of the bowel is unusual in children, an underlying predisposing condition should be sought in affected cases. PMID- 2553913 TI - 5-year evaluation of durapatite ceramic alloplastic implants in periodontal osseous defects. AB - Six patients who participated in a clinical study comparing the response of periodontal osseous defects to either grafting with Periograf (durapatite) hydroxylapatite (HA) ceramic or debridement alone (DEBR) were followed under an active maintenance program for 5 years. Mean values for gingival recession and attachment levels remained essentially stable following either treatment over the 5 year period. However, mean probeable pocket depths shifted from being significantly (P less than 0.05) shallower for DEBR sites at 6 months and 1 year to being significantly shallower for HA sites at 5 years. The probing pocket depth change for grafted sites was steady and the change from presurgical values was significantly greater than the change for DEBR areas, which became about 1.5 mm deeper over the 5 year postsurgical period. Intrapatient comparisons showed that recession, attachment gain, and pocket depth decrease were most frequently greater for the Periograf-treated sites. Assessment of the pattern of clinical changes during the 5 year postsurgical period demonstrated that the attachment level of grafted sites improved or stayed the same 86% of the time compared to only 62% stability or improvement in the DEBR only sites. In fact, 38% of the DEBR sites were worse at 5 years than at the time of surgery, a 3 times greater failure rate than that found in the HA-grafted sites. Pocket depth measurements showed that 98% of the Periograf-treated sites were better or the same as presurgically (2% worse), while only 80% of DEBR sites showed positive results and 20% had deeper pockets than at the time of surgery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553914 TI - Effects of isoproterenol on synaptic ribbons in pinealocytes of the rat and C57BL/6J mouse. AB - Synaptic ribbons (SR) in melatonin-deficient pinealocytes of the C57BL/6J mouse were quantitatively compared to SR in pinealocytes of the rat after beta adrenergic receptor activation by isoproterenol. Two populations of SR comprising synaptic spherules (SRsp) and synaptic rods (SRr) were described in both the mouse and the rat, but species differences existed in the ratio of SRr to SRsp. Isoproterenol caused a significant increase in frequency of SR of the rat but had little or no effect on SR populations in the mouse. It is unlikely that beta adrenergic receptors are absent on mouse pinealocytes or were not activated since isoproterenol elevated plasma renin concentrations indicating activation of beta adrenergic receptors. Furthermore the pineal of both species receives heavy sympathetic input. These findings indicate that the role and regulation of pinealocyte SR are complex and are functionally linked to beta-adrenergic receptors as well as other mechanisms related to the production of melatonin. PMID- 2553915 TI - [Differential effect of sodium bicarbonate on disposition of sulfadimethoxine and sulfisoxazole in rabbits]. AB - The orally co-administered sodium bicarbonate significantly enhanced the blood concentration of sulfadimethoxine at the early stage after oral administration to rabbits, by increasing its intestinal absorption. On the other hand, the sodium bicarbonate significantly reduced the blood concentration of sulfisoxazole at the elimination phase after oral administration to rabbits, by increasing its urinary excretion. The fact that sodium bicarbonate exhibits different effects in the disposition of these two sulfonamides is an interesting example to gain a better understanding for the complexity of drug interaction. PMID- 2553916 TI - Egestion of asbestos fibers in Tetrahymena results in early morphological abnormalities. A step in the induction of heterogeneous cell lines? AB - In Tetrahymena populations exposed to crocidolite asbestos fibers, many cells develop morphological abnormalities within 1-2 hours. The abnormalities are mainly large or small protrusions or indentations, or flattened parts of the cell surface and most often located in the posterior part of the cell. They are formed repeatedly in all cells but are also continuously repaired so that the fraction of cells affected represents an equilibrium between these two processes. Their formation is connected with egestion of the large bundles of fibers formed by phagocytosis. Such effects of egestion of fibers do not seem to have been reported previously. Egestion of a bundle of fibers is much slower than for other types of undigestible residues. In contrast to normal exocytosis occurring invariably at the cytoproct, egestion of asbestos often occurs in the posterior part of the cell outside the cytoproct. To my knowledge this is the first reported case of either very slow or extra-cytoproctal egestion in Tetrahymena. Cells with large abnormalities have a greater tendency to develop into "early heterogeneous" cells than the average abnormal cell. Some of these give rise to hereditarily stable heterogeneous cell lines of Tetrahymena. The morphological abnormalities are probably caused by mechanical action of the crocidolite fibers resulting in local damage of the cytoskeletal elements responsible for normal cell shape. The heterogenous cell lines may arise when cellular structures carrying non-genic cytotactically inherited information are modified. The relevance of these ideas to the induction of cancer by asbestos is briefly discussed. PMID- 2553917 TI - Effects of chronic morphine administration on the mu and delta opioid responses in the CA1 region of the rat hippocampus. AB - Extracellular recording of population spike amplitudes in the hippocampal CA1 region were compared in slices from control and chronically morphine-treated rats. Morphine treatment resulted in a decrease in the maximal excitation produced by both mu and delta selective agonists, [N-MePhe3,D-Pro4]-morphiceptin and [D-Pen2,L-Pen5]-enkephalin. The opioid antagonist naloxone did not produce apparent signs of withdrawal in hippocampal slices from morphine-treated rats as shown by a lack of change in the evoked population spike in the presence of 500 nM naloxone. Brain slices from morphine-treated rats that were maintained in the absence of morphine showed signs of tolerance reversal when monitored over an 8 hr period after dissection. If morphine-tolerant slices were maintained in vitro in the presence of 5 microM morphine (the concentration found by high-performance liquid chromatography to be present in the cerebrospinal fluid of morphine treated rats), there was no significant reversal of tolerance. Furchgott analysis of the [N-MePhe3,D-Pro4]-morphiceptin concentration-response curve shifts induced by the irreversible opioid receptor antagonist beta-chlornaltrexamine revealed an apparent 50% spare receptor reserve for the mu selective agonist in slices from drug-naive rats. beta-Chlornaltrexamine (20 nM) treatment and chronic morphine exposure resulted in a similar reduction in the maximal response to [N-MePhe3,D Pro4]morphiceptin. This observation indicates that the development of morphine tolerance resulted in an elimination of the spare opioid receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553918 TI - Neuroendocrine evidence for denervation supersensitivity of serotonin receptors: effects of the 5-HT agonist RU 24969 on corticotropin, corticosterone, prolactin and renin secretion. AB - Serotonergic stimulation can increase the secretion of several hormones through the involvement of different serotonin (5-HT) receptor subtypes. RU 24969, a 5-HT agonist with highest affinity at 5-HT1A and 5-HT1B receptors, increased plasma renin activity (PRA) and plasma renin concentration (PRC) as well as plasma corticosterone and prolactin concentrations in a dose-dependent manner. Inasmuch as 5-HT2 receptors mediate the serotonergic stimulation of renin secretion, we examined the ability of two selective 5-HT2 antagonists, ritanserin and LY53857, to inhibit the neuroendocrine effects of RU 24969. To determine whether the 5-HT receptors which are involved in the stimulation of these hormones are pre- or postsynaptic, RU 24969 was also injected to rats whose brain serotonergic neurons were chemically destroyed by i.c.v. injection of 5,7-dihydroxytryptamine. Both ritanserin and LY53857 blocked the effect of RU 24969 on PRA and PRC, but did not inhibit the RU 24969-induced elevation in plasma corticosterone concentrations. Ritanserin did not inhibit the effect of RU 24969 on prolactin levels, but LY53857 produced a partial inhibition of the RU 24969-induced elevation of prolactin concentrations. In rats with chemical lesions of serotonergic neurons the dose-response curves of RU 24969 for PRA and PRC as well as corticotropin, corticosterone and prolactin shifted to the left, suggesting functional up regulation of postsynaptic 5-HT receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553919 TI - Reinforcing effect of alfentanil is mediated by mu opioid receptors: apparent pA2 analysis. AB - Apparent pA2 analysis was used to determine whether the short-duration opioid agonist, alfentanil, acts at mu receptors in the positive reinforcement of operant behavior in the rhesus monkey. In test sessions a red light signaled the availability of alfentanil injections. If a monkey pressed a response lever 30 times, a specific dose of alfentanil was injected i.v., and the red light was extinguished for 10 min. This cycle could be repeated for up to 130 min, the maximum length of a session. Between successive test sessions at least three maintenance sessions were held; in these sessions injections of 0.32 mg/kg/injection of codeine were made available. The dose of alfentanil was changed from one test session to the next, and dose-dependent changes in rates of responding resulted. Rates reached 3.05 responses/sec at 0.010 mg/kg/injection, the highest dose tested. The opioid antagonist, quadazocine, produced dose dependent, parallel shifts to the right in the alfentanil dose-response curve. In Schild Plot analysis the regression line fit to the antagonism data had a slope of -1.1; the apparent pA2 value for quadazocine was 7.6. This value was close to apparent pA2 values obtained with mu agonists in studies of other behavioral effects of opioids, but distinct from values obtained with kappa agonists in those studies. Thus, it is likely that mu receptors mediate the positive reinforcing effect of alfentanil. PMID- 2553920 TI - Kappa opiate receptor multiplicity: evidence for two U50,488-sensitive kappa 1 subtypes and a novel kappa 3 subtype. AB - Kappa receptor multiplicity is a complex area. We now present evidence from binding studies suggesting the existence of four kappa receptor subtypes. The guinea pig cerebellum contains high levels of U50,488-sensitive, or kappa 1, receptors. Kappa opiates (U50,488, tifluadom, Mr2034, Mr2266 and Win44,441) compete [3H]ethylketocyclazocine binding to kappa 1 receptors with kappa, values under 10 nM and Hill coefficients of approximately one, as does dynorphin A (kappa 1, 0.27 +/- 0.05 nM; Hill coefficient, 0.83 +/- 0.20, n = 4). However, competition studies with dynorphin B yield a Hill coefficient of 0.46 +/- 0.03 (n = 5) and nonlinear regression analysis of the competition curve is best fit by two sites. alpha-Neoendorphin Neoendorphin competition curves (Hill coefficient, 0.46 +/- 0.07; n = 3) also were best fit with two components. Competition studies with both alpha-neoendorphin and dynorphin B together suggest that both compounds label the same site with high affinity. Similar results were obtained using [3H]U69,593. Dynorphin B and alpha-neoendorphin competed binding with Hill coefficients of 0.45 +/- 0.04 (n = 3) and 0.59 +/- 0.09 (n = 3), respectively. These data suggest two subtypes of kappa 1 receptors in the guinea pig cerebellum: kappa 1a and kappa 1b. Classical kappa opiates and dynorphin A have high affinity for both subtypes whereas dynorphin B and alpha-neoendorphin label kappa 1b over 50-fold more potently than kappa 1a sites. [3H]Naloxone benzoylhydrazone [( 3H]NalBzoH) labels a novel, U50,488-insensitive kappa receptor subtype, kappa 3, in membranes from calf striatum, rat and mouse brain. We now have developed a relatively selective assay in calf striatum.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553921 TI - Pharmacological actions of a novel mixed opiate agonist/antagonist: naloxone benzoylhydrazone. AB - NalBzoH (6-desoxy-6-benzoylhydrazido-N-allyl-14-hydroxydihydronomorphin one) is a novel opiate with potent actions at both mu and kappa receptors. Analgesic studies in mice examining increasing doses of NalBzoH with a fixed dose of morphine revealed a biphasic curve. NalBzoH at doses as low as 1 microgram/kg partially antagonized morphine analgesia. Higher NalBzoH doses continued to inhibit morphine analgesia in a dose-dependent manner, with the 1-mg/kg dose antagonizing completely morphine analgesia. As the NalBzoH dose increased beyond 1 mg/kg analgesia returned. NalBzoH also prduced a similar analgesic response when administered alone in mice and also was active in rats. NalBzoH had excellent p.o. activity, with an analgesic potency in mice equivalent to s.c. administration. Naloxone reversed NalBzoH analgesia far less effectively than morphine analgesia. In contrast, Win44,441 antagonized both morphine and NalBzoH analgesia with a similar potency, consistent with a kappa mechanism for NalBzoH analgesia. Repeated administration of NalBzoH resulted in tolerance. There was no analgesic cross-tolerance between NalBzoH and either morphine or the kappa 1 selective agent U50,488H, implying a selective kappa 3 mechanism of analgesia. In addition to blocking morphine analgesia, low doses of NalBzoH also partially reversed the inhibition of gastrointestinal transit in mice produced by morphine, antagonized completely morphine lethality and precipitated withdrawal in morphine dependent mice, confirming its antagonist activity in mu receptors. The duration of NalBzoH's kappa and mu actions differed dramatically. In mice, analgesia typically lasted less than 2 hr whereas the same NalBzoH dose antagonized completely morphine analgesia, a mu action, for 16 hr. Full sensitivity to morphine did not return for 32 hr.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553922 TI - SK&F 104353: selective antagonism of peptidoleukotrine-induced changes in electrolyte transport by rat ileal mucosa in vitro. AB - Effects of the peptidoleukotriene receptor antagonist, SK&F 104353, were examined in vitro using stripped rat ileal mucosa in Ussing chambers. Changes in short circuit current (Isc), transepithelial conductance (G1) and unidirectional and net Na+ and Cl fluxes measured in the absence or presence of SK&F 104353 (1 microM) revealed that serosal addition produced a decrease in net Na+ absorption and serosal-to-mucosal Cl- flux. Serosal addition of leukotriene (LT)D4 (10 muM) or LTE4 (10 microM) elicited transient increases in Isc and sustained decreases in G1 which were antagonized by serosal addition of SK&F 104353 in a concentration-dependent manner. Mucosal addition of SK&F 104353 (1 microM) also reduced the increase in Isc elicited by LTD4 (10 microM). LTD4 (10 microM) decreased unidirectional and net Na+ and Cl- fluxes along with G1 in the steady state. All of these changes were abolished by pretreatment with SK&F 104353 (1 microM). The intestinal secretagogues prostaglandin F2 alpha, histamine, serotonin, substance P and the thromboxane mimic, U46619, produced changes in Isc qualitatively similar to those of LTD4. However, the transient increase in Isc produced by these secretagogues was not antagonized by SK&F 104353 (1 microM). Additionally, lysbradykinin- and prostaglandin E1-induced increases in Isc were not antagonized by SK&F 104353 (1 microM). Stimulation with histamine (10 microM) or pretreatment with LTB4 (5 microM) did not alter the increase in Isc or decrease in Gt produced by the subsequent addition of LTD4 (10 microM). Pretreatment of the tissues with mepyramine (10 microM) also did not reduce the increase in Isc elicited by LTD4 (10 microM), although it inhibited completely the increase in Isc produced by histamine (10 microM).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553923 TI - Pharmacologic experiments on the interaction between crotoxin and the mammalian neuromuscular junction. AB - Crotoxin and its two subunits were tested for their neuromuscular blocking activity on the phrenic nerve-hemidiaphragm preparation. Two types of experimental paradigms were used, the first of which separated the toxin binding step from subsequent events in paralysis and the second of which did not. In both paradigms the toxin produced concentration-dependent blockade of transmission. However, the results with low concentrations were variable, and in some cases complete neuromuscular blockade did not develop. The isolated acidic and basic subunits possessed little toxicity. In experiments designed to characterize binding, the intact toxin displayed the following properties: 1) the apparent half-time for tissue association was about 22 min; 2) binding was not affected by low temperature, the presence or absence of nerve stimulation and the substitution of strontium for calcium; and 3) when binding was allowed to go to completion, reversibility was negligible. Pretreatment of tissues with the isolated subunits of crotoxin did not enhance or inhibit the binding of the parent molecule. Modification of one histidine residue in the isolated basic subunit, followed by reconstitution with unmodified acidic subunit, generated a molecule that possessed only about 10% of the neurotoxicity of the native toxin. The modified toxin could not be used to antagonize binding of the native toxin. Both polyclonal and monoclonal antibodies were generated that neutralized the biologic activity of crotoxin. In experiments that separated the binding step from later events in paralysis, the polyclonal preparation continued to locate and partially neutralize tissue-bound toxin. In experiments that initiated events that follow binding, polyclonal antibodies were progressively less effective with time in neutralizing toxin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553924 TI - Opioid-induced prejunctional inhibition of vasoconstriction in the rabbit ear artery: alpha-2 adrenoceptor activation and external calcium. AB - Inhibition of norepinephrine release by opioid agonists is inversely related to the stimulation train length. The possible interaction between activation of prejunctional alpha-2 adrenergic receptors and the release-inhibiting opioid receptors as well as the effect of changes in Ca++ entry into adrenergic varicosities during repetitive stimulation were investigated by recording vasoconstriction of the rabbit ear artery perfused in vitro. Neither the activation of the alpha-2 adrenoceptor-mediated negative feedback by clonidine nor its inhibition by yohimbine altered the neuroinhibitory potency for dynorphin 1-13 or Met-enkephalin at any stimulus train length. Experimental conditions known to increase the entry of calcium into the varicosities mimicked the effect of the increase in the stimulation train length on the modulation of norepinephrine release by opioids. Increasing the extracellular calcium concentration (from 1.6-5 or 8 mM) diminished the inhibitory effect of opioids and tended to abolish the dependence on stimulation train length. Conversely, lowering the calcium concentration (from 1.6-1 mM) increased the inhibition by opioids and enhanced the dependence on train length. These results do not suggest a direct interaction between activation of opioid receptors and alpha-2 adrenoceptors. Rather, they indicate the role of opioid receptor activation in a primary modulation of Ca++ influx which becomes masked by the high levels of axoplasmic calcium which are achieved during a continued stimulation train. PMID- 2553925 TI - Antagonism to the actions of platelet activating factor by a nonpsychoactive cannabinoid. AB - A recent report from our laboratory gave evidence showing that tetrahydrocannabinol (THC)-7-oic acid has antinociceptive activity in the mouse. We also pointed out that this substance, which is a major metabolite of THC in most species including humans, is probably responsible for the well known analgesic properties of the parent drug. The present report contains findings that suggest THC-7-oic acid may have considerable activity as an antagonist to platelet activating factor, which may also explain the known properties of THC as a bronchodilator, antipyretic and antirheumatic agent. In the mouse ear edema test, THC-7-oic acid appeared to be about as efficacious as phenidone; however, its potency was less than either phenidone or indomethacin. When compared with the parent drug, THC, in the platelet activating factor-induced paw edema assay, it acted more quickly and produced a greater reduction of edema. This is consistent with the possibility that THC must be metabolized to the 7-oic acid for activity to be seen. Its activity in preventing platelet activating factor induced mortality was comparable to naproxen. In vitro studies suggest that THC-7 oic acid can inhibit both cyclooxygenase and 5-lipoxygenase activities in intact cells. PMID- 2553926 TI - A mu-specific opioid peptide agonist increases excitability of pyramidal neurons in untreated and receptor up-regulated hippocampus. AB - The rat hippocampus contains the major types of opioid receptors, delta, mu, and kappa, as determined by autoradiographic and membrane binding analyses. Chronic exposure to excessive amounts of opioid antagonists results in a doubling of the number of binding sites. However, the direct electrophysiological significance of this increased number of opioid receptors in the central nervous system remains uncharacterized. We examined the effects of an opioid peptide with high affinity and high specificity for mu receptors, DAMGO (D-ala2-mePhe4-gly-ol5 enkephalin), under normal conditions and after 1 or 2 weeks of continuous infusion of the opiate antagonist naltrexone. Chronic infusion of naltrexone administered to the whole animal resulted in significant up-regulation (71%) of mu opioid receptors in the rat hippocampus. Slices of the hippocampus were perfused with artificial cerebrospinal fluid while recording population spikes in stratum pyramidale, excitatory postsynaptic potentials in stratum radiatum and while stimulating afferents in the Schaffer collaterals. Superfusion of slices with DAMGO produced a concentration-dependent increase in the amplitude of population spikes. No significant change was observed in the simultaneously recorded excitatory postsynaptic potential slope. This selective increase in population spike amplitude led to a leftward shift (19%) in the derived input-output curve. In addition, DAMGO superfusion produced extra spiking at higher stimulus intensities. Naltrexone reversed the DAMGO-induced increase in excitability, as well as prevented additional spikes. DAMGO superfusion of slices taken from chronically treated rats produced a much greater shift (42%) in the input-output curve than it did in untreated controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553927 TI - Aging does not alter brain muscarinic receptor-mediated phosphoinositide hydrolysis and its inhibition by phorbol esters, tetrodotoxin and receptor desensitization. AB - The effects of aging on muscarinic receptor-mediated phosphoinositide (PI) hydrolysis in the brain were investigated in Fischer-344 rats. Oxotremorine-M stimulated this response to the same magnitude in young and old rats in the cerebral cortex, striatum, hippocampus, thalamus, hypothalamus and cerebellum, although maximal stimulation varied among brain regions within each age group. The PI response was also equally potentiated by elevated K+ or suppressed by tetrodotoxin in both age groups. In addition, a phorbol ester attenuated muscarinic receptor-mediated PI hydrolysis in the cerebral cortex to the same extent in young and aged rats. Moreover, preincubation with oxotremorine-M resulted in a similar down-regulation of cell-surface receptors and desensitization of receptor function regardless of age. Therefore, under our experimental conditions, PI hydrolysis in response to activation of brain muscarinic receptors does not appear to be sensitive to aging-related alterations. PMID- 2553928 TI - Indirect involvement of delta opioid receptors in cholecystokinin octapeptide induced analgesia in mice. AB - Sulfated cholecystokinin octapeptide (CCK-8; Asp-Tyr-SO3H-Met-Gly-Trp-Met-Asp-Phe NH2) produced analgesia in mice when administered i.c.v. and tested in the acetic acid-induced writhing assay. The ED50 was found to be 0.03 nmol/mouse which was about 3, 24 and 714 times more potent than morphine. [D-Pen2,D-Pen5]enkephalin and U50,488H [trans-(+/-)-3,4-dichloro-N-methyl-N-[2-(1-pyrolidinyl)cyclohexyl] benzeneacetamidel], respectively. When administered i.t., CCK-8 produced partial analgesia of up to 22 to 23% at low doses ranging from 15 to 60 ng/mouse and hyperalgesia at doses over 120 ng/mouse. Naloxone, an opioid antagonist, inhibited the analgesia induced by CCK-8 (i.c.v. and i.t.) but potentiated hyperalgesia induced by CCK-8 (i.t.). Apparent pA2 value for CCK-8 (i.c.v.) against naloxone (s.c.) was 5.88 which was significantly different from those for morphine-naloxone and U50,488H-naloxone but was not significantly different from that for [D-Pen2,D-Pen5]enkephalin-naloxone. Studies using highly selective opioid antagonists showed that CCK-8-induced analgesia was significantly antagonized by the delta receptor antagonist, ICI154,129 [(Allyl)2-Tyr-gly-gly psi-(CH2S)-Phe-Leu] but not by beta-funaltrexamine, a highly selective mu receptor antagonist or nor-binaltorphimine, a highly selective kappa receptor antagonist. Opioid receptor binding study using [3H]-[D-Ala2,D-Leu5]enkephalin (+unlabeled [D-Ala2,MePhe4,Gly-ol5]enkephalin) in mouse brain membrane preparations revealed that there were no changes in the maximum binding or Kd of delta opioid binding sites in the presence of CCK-8 (1 microM) in vitro.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553929 TI - Neutrophils delay functional recovery of the post-hypoxic heart of the rabbit. AB - A postischemic contractile dysfunction termed myocardial stunning has been described in vivo and is attributed, in part, to the generation of oxygen-derived free radicals and the presence of neutrophils. An analogous contractile derangement occurs in the posthypoxic heart in vitro. This study determined the role of neutrophils in hypoxia/reoxygenation-induced cardiac dysfunction in the isolated buffer-perfused rabbit heart utilizing a recirculating system with or without neutrophils present. In control hearts perfused with a neutrophil-free buffer, reoxygenation after 20 min of hypoxia was associated with a slow recovery of contractility which returned to prehypoxic values by 30 to 45 min. Although perfusion with buffer-containing neutrophils did not affect the hypoxia-induced decrease in myocardial contractility, the recovery of contractile function during subsequent reoxygenation was significantly diminished (P less than .01 vs. control), remaining depressed by 30 to 35% at 45 min. The myocardial neutrophil content increased approximately 2-fold in response to hypoxia and reoxygenation, as assessed using 51Cr-labeled neutrophils. The deleterious effects of neutrophil perfusion on cardiac function could not be attributed to neutrophil-mediated plugging of coronary vessels or enhanced myocellular damage. These results support the concept that neutrophils contribute to the cardiac dysfunction described in this model. PMID- 2553930 TI - Effects of competitive and noncompetitive N-methyl-D-aspartate (NMDA) antagonists in rats trained to discriminate NMDA from saline. AB - Competitive and noncompetitive N-methyl-D-aspartate (NMDA) antagonists and other central nervous system depressants were assessed for their ability to antagonize the discriminative stimulus effects of NMDA in rats trained under a standard two lever fixed ratio schedule of food reinforcement. The competitive NMDA antagonists, 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonate and NPC 12626 [2 amino-4,5-(1,2-cyclohexyl)-7-phosphonoheptanoate], dose-dependently antagonized NMDA-lever selection at doses that did not affect rates of responding. Conversely, the noncompetitive NMDA antagonists, phencyclidine, MK-801 [(+)-5 methyl-10,11-dihydro-5H-dibenzo(a,d)cyclohepten-5,10-imine maleate] and (+)-N allylnormetazocine, as well as pentobarbital and diazepa, all reduced response rates dose-dependently without antagonism of NMDA-lever responding. In stimulus generalization tests, NPC 12626 and 3-(2-carboxypiperazin-4-yl)propyl-1 phosphonate at doses higher than those required to antagonize NMDA, often elicited NMDA-lever responding. The mechanisms underlying the similarities in the interoceptive stimuli produced by NMDA and its competitive antagonists remain to be determined. These results indicate that although competitive NMDA antagonists antagonize effects of NMDA without concomitant behavioral disruption, noncompetitive NMDA antagonists and central nervous system depressants are behaviorally disruptive at doses that do not antagonize NMDA. The results provide further evidence for differences in the behavioral profiles of competitive and noncompetitive NMDA antagonists. PMID- 2553931 TI - Alpha-2A and alpha-2B adrenergic receptor subtypes: attenuation of cyclic AMP production in cell lines containing only one receptor subtype. AB - At least two subtypes of alpha-2 adrenergic receptors have been identified on the basis of antagonist affinities as determined mainly by radioligand binding assays. The human platelet and the HT29 human colonic adenocarcinoma cell contain alpha-2A adrenergic receptors, whereas the neonatal rat lung and the NG108 neuroblastoma X glioma hybrid cell contain the alpha-2B adrenergic receptor. Using the attenuation of the cyclic AMP accumulation as a functional assay, the affinities of various antagonists for the alpha-2 adrenergic receptor were determined in HT29 and NG108 cell lines. Dose-response curves to 5-bromo-6-(2 imidazoline-2-yl-amino)quinoxaline (UK 14,304) an alpha-2 adrenergic agonist, were generated in the absence and presence of three concentrations of various antagonists. Schild regressions were used to determine pA2 values and then dissociation constants (KB value) were calculated. Whereas phentolamine and yohimbine were equipotent at the receptor in the two cell lines, 2-(2,4-(O methoxyphenyl)-piper-azin-1-yl)ethyl-4,4-dimethyl-1,3-(2 H,4H)- isoquinolindione (ARC-239) and prazosin were 100- and 30-fold more potent in the NG108 cell line than in the HT29 cell. These potency ratios determined from functional experiments are the same as those obtained from radioligand binding experiments. These functional data are consistent with the previous and more extensive binding data, and thus support the existence and definition of alpha-2A and alpha-2B adrenergic receptor subtypes. PMID- 2553932 TI - Amiodarone blocks calcium current in single guinea pig ventricular myocytes. AB - Ca++ current (lca) block by amiodarone and the underlying mechanisms thereof were investigated in guinea pig single ventricular myocytes using the single suction pipette whole cell voltage clamp method. The dose-response curve revealed a 1:1 stoichiometry for binding of amiodarone to its receptor with an apparent dissociation constant of 5.8 microM in the resting state. Amiodarone, 5 microM did not significantly alter the time course of ICa decay, but did shift the steady-state inactivation curve for lca in the hyperpolarizing direction by 9.2 +/- 3.1 mV. Development of block at depolarized potentials was voltage-dependent between -20 and 10 mV with time constants of 112 +/- 33 and 755 +/- 212 msec at 10 mV. In the presence of 0.2 microM amiodarone, recovery from inactivation was fitted by a double exponential most likely indicating rapid recovery of the drug free Ca++ channels and slow recovery of the drug-associated Ca++ channels with time constants of 44 +/- 12 and 108 +/- 403 msec, respectively, at -80 mV. The proportion of the current recovering via the slow phase was 36 +/- 7%. By using this value, we estimated the dissociation constant in the inactivated state to be 0.36 microM. Amiodarone's marked use-dependent block of lca is explicable in terms of its high affinity for, and slow dissociation from, Ca++ channels in the inactivated state. These results suggest that amiodarone blocks lca in both the resting and inactivated states.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553933 TI - Structure-activity profile of calcitonin gene-related peptide in peripheral and brain tissues. Evidence for receptor multiplicity. AB - The radioligand binding characteristics and in vitro biological activities of human calcitonin gene-related peptide (hCGRP) homologs, analogs and fragments have been tested in a variety of preparations in order to determine the structural requirements of the calcitonin gene-related peptide (CGRP) receptor. The ligand selectivity profile observed in this study suggests that [125I] hCGRP binding sites present in rat brain and spleen are similar. The high affinity of the analogs [Cys(ACM)2.7]hCGRP and cyclo2.7 [Asp2, Lys7]hCGRP indicates that the disulfide bridge between positions 2 and 7 is not essential for the binding of the ligand to the CGRP receptor. On the other hand, the lack of affinity of various N-terminal fragments in the binding assays demonstrates the importance of amino acids 9-37 in maintaining appropriate receptor recognition. The existence of CGRP receptor subtypes is supported by data obtained in various in vitro bioassays. For example, the linear analog, [Cys(ACM)2.7]hCGRP, which retained high potency in the rat vas deferens bioassay displayed no agonistic activity in the right and left guinea pig atrial preparations. Additionally, the C-terminal fragment hCGRP 12-37 possessed some antagonist properties in atrial but not vas deferens preparations. Thus, it appears that it is possible to differentiate between CGRP receptor subtypes on the basis of differential agonistic (e.g.[Cys(ACM)2.7]hCGRP) and antagonistic (e.g., hCGRP 12-37) potencies in various isolated tissues. Moreover, the weak but significant antagonistic properties of hCGRP 12-37 may be most useful in order to design more potent CGRP receptor antagonists. PMID- 2553934 TI - Two different human papillomavirus (HPV) types associated with oral mucosal lesions in an HIV-seropositive man. AB - Different types of Human papillomaviruses (HPV) are associated with a variety of oral lesions. So far, HPV types 1, 2, 4, 6, 7, 11, 13, 16, 18, 32 and 57 have been identified in oral lesions. Immunosuppression predisposes oral mucosa to clinical manifestation of different virus infections including HPV. We describe here a 30-year-old HIV-positive and immunosuppressed man, who had suffered from oral lesions for a few months. On clinical examination, a nodular elevation was detected on the lower lip, and white keratotic areas were present on buccal mucosa bilaterally. A biopsy from the lip revealed the presence of acanthosis with a prominent granular cell layer as well as hyperparakeratosis. A biopsy from the buccal lesion showed a comparatively much flatter lesion with merely basal cell hyperplasia associated with hyperparakeratosis. Koilocytosis was a characteristic feature in both biopsies. In Southern blot hybridization, both lesions hybridized with a probe cocktail comprising HPV 6, 11, 16, 18, 31 and 33 DNA under low stringency. Under high stringency, the lip lesion proved to contain HPV 7 DNA, which also confirmed by in situ hybridization. The buccal lesion was weakly positive by Southern blot with HPV 11 and 13 probes hybridized under stringent conditions, but the restriction patterns with Pst I and Bam HI did not fit with those of any of the 57 HPV types known so far. In situ hybridizations with HPV 11 and HPV 13 probes were negative. Cloning of this 'new' HPV type is currently under way. PMID- 2553935 TI - Circular DNA of Entamoeba histolytica encodes ribosomal RNA. AB - The presence of repeated DNA sequences encoding RNA in Entamoeba histolytica has been reported. In the present study we demonstrate by agarose gel electrophoresis. DNase digestion and electron microscopic analysis that these genes are located on extrachromosomal circular DNA molecules with an approximate size of 26 kb. Detection of replication intermediates suggests the episomal nature of these molecules. Amplified, extrachromosomal rRNA genes appear to be a common feature among the lower eukaryotes, occurring more commonly as linear molecules and less commonly as circles. Entamoeba histolytica is 1 of the few organisms studied in which rRNA genes are located predominantly on extrachromosomal circles. PMID- 2553936 TI - Breast cancer as seen in King Fahd University Hospital, Al-Khobar, 1983-1986. AB - Breast cancer is the most common female cancer in Saudi Arabia as reported in several studies based on data from specialized centres. The numbers seen in most general hospitals, however, are low, making it difficult to carry out comprehensive clinicopathological studies of the disease. We present the results of a hospital-based study which confirm the low hospital case incidence. The disease appears to be commoner in a younger age group, with a higher occurrence of medullary carcinoma compared with studies carried out elsewhere. These findings emphasize the need for a national population-based study as the best method for defining the problem of breast cancer in the Kingdom of Saudi Arabia. PMID- 2553937 TI - Carcinoma of breast presenting as gastric cancer: diagnosis by immunoperoxidase staining patterns. PMID- 2553938 TI - Effect of static magnetic field on the induction of chromosome aberrations by 4.9 MeV protons and 23 MeV alpha particles. AB - The effect of static magnetic field applied during in vitro exposure to 4.9 MeV protons and 23 MeV alpha particles on the induction of chromosome aberrations in human peripheral blood lymphocytes was studied. The mean frequency of dicentrics in cells irradiated in magnetic field of a magnitude about one Tesla tended to be higher than that irradiated in the absence of magnetic field for both protons and alpha particles, and the analysis of dose-effect relationships indicated that for protons the magnetic field significantly affected the dose-effect relationship. The intercellular distribution of chromosome aberrations, one of the measures of radiation quality, was not significantly influenced by the magnetic field. PMID- 2553939 TI - General practice or primary health care? PMID- 2553940 TI - Dietary treatment of infant colic: a double-blind study. AB - The possible role of cows milk intolerance in the aetiology of infant colic was evaluated in 19 babies presenting to their health visitor or general practitioner in one town. Over a three week period a standard modified cows milk formula was compared with a soya milk formula on a double blind basis. The duration of colic symptoms was significantly reduced during the week on soya milk (P less than 0.01), with 11 out of 19 babies fulfilling the diagnostic criteria for cows milk intolerance. Four babies whose symptoms failed to improve either spontaneously or with soya milk were given a hydrolysed protein milk with a positive response in two, confirmed by challenge testing. Thus in 13 out of 19 babies (68%) the symptoms of infant colic resolved almost completely with dietary change. PMID- 2553941 TI - Preventive care card for general practice. AB - A new preventive care card for use in general practice has been designed and produced by a working party of general practitioners from the local medical committees of Devon and Cornwall and the Tamar faculty of the Royal College of General Practitioners. The new card, which is described in this paper, is proposed as a replacement for forms FP7A and FP8A in general practice records. PMID- 2553942 TI - Elderly peoples' experiences of discharge from hospital. AB - In a study of patients' perceptions of the transition from hospital to the community, 115 elderly people registered with a central London group practice were interviewed shortly after they arrived home. Many patients received little notice of discharge, a third being told on the day they left the hospital. A third felt they had been discharged too soon and those living alone were significantly less likely to return to a heated home containing basic items of food. Seventy-seven patients, including 80% of those living alone were visited by family, friends or professionals within three days of coming home. Eighty-six per cent of non-professional visitors were women. Several of the elderly couples appeared to be under considerable stress and not all individuals were receiving the help they considered most appropriate to their needs. Many of the problems identified were due to poor communication between practice, hospital and patients. We suggest several measures aimed at improving the quality of that communication, so as to ensure that available resources can be mobilized to support this vulnerable group of people. PMID- 2553943 TI - Routine treatment of cutaneous warts: a questionnaire survey of general practitioners. AB - A postal questionnaire was sent to 185 general practitioners to assess their approach to cutaneous warts and their views on the future development of the routine wart treatment service; 159 (85.9%) replied. A wide range of treatments were offered and most patients were given some treatment. The main reasons respondents gave for referring patients to hospital were failure of wart paints (73.6%) and lack of availability of liquid nitrogen (70.4%). Most general practitioners (74.2%) believed that dermatologists should spend less than 5% of their time treating warts. Many general practitioners (61.6%) wanted a practice based wart clinic offering cryotherapy and 30.8% would like to refer directly to a hospital clinic run by a nurse. A practice clinic was more popular with general practitioners who have a treatment room nurse (P less than 0.01). Most seemed to appreciate the need for training to use liquid nitrogen. We conclude that general practitioners are keen to use cryotherapy and we argue that hospital management should provide the necessary resources for running a community-based service. PMID- 2553944 TI - Extending appointment length--the effect in one practice. AB - The effects of extending appointment length from 7.5 to 10.0 minutes in the evening surgeries of two general practitioners in one general practice were examined. The mean consultation length increased from 7.4 to 8.4 minutes. Most of this change appeared to be due to a small increase in the length of shorter consultations and was true for all age groups. The time that patients had to wait did not decrease and for one doctor actually increased. However, the results suggest that with longer appointments there is more discussion of lifestyle factors and more screening activity. PMID- 2553946 TI - Activity, audit and accountability. PMID- 2553945 TI - Management of chronic (post-viral) fatigue syndrome. AB - Simple rehabilitative strategies are proposed to help patients with the chronic fatigue syndrome. A model is outlined of an acute illness giving way to a chronic fatigue state in which symptoms are perpetuated by a cycle of inactivity, deterioration in exercise tolerance and further symptoms. This is compounded by the depressive illness that is often part of the syndrome. The result is a self perpetuating cycle of exercise avoidance. Effective treatment depends upon an understanding of the interaction between physical and psychological factors. Cognitive behavioural therapy is suggested. Cognitive therapy helps the patient understand how genuine symptoms arise from the frequent combination of physical inactivity and depression, rather than continuing infection, while a behavioural approach enables the treatment of avoidance behaviour and a gradual return to normal physical activity. PMID- 2553947 TI - Timing and purpose of the MRCGP examination. AB - There is increasing disquiet among course organizers about trainees who sit the MRCGP examination before completing vocational training. An alternative arrangement is proposed for trainees to undergo continuous assessment during vocational training with a more clinical end-point assessment than the current MRCGP examination. Career progression for general practitioners would then be available in the form of the MRCGP examination after a number of years in practice and the FRCGP by assessment at a later stage. It is suggested that the government might wish to reward evidence of a commitment to continuing education. PMID- 2553948 TI - Patients' knowledge and attitudes to HIV and AIDS. PMID- 2553949 TI - Knowledge of health issues among Asian and white schoolchildren in Glasgow. PMID- 2553950 TI - Incidence of mumps. PMID- 2553951 TI - Breast self examination. PMID- 2553952 TI - Clinical psychologists in general practice. PMID- 2553953 TI - James Mackenzie lecture 1988. Everyone's business--everyone's responsibility. PMID- 2553954 TI - Control of multiple transducing systems by LH which results in the modulation of adenylate cyclase, protein kinase C, lipoxygenases and cyclooxygenases. PMID- 2553955 TI - Control of the steroidogenic machinery of the human trophoblast by cyclic AMP. AB - Human cytotrophoblasts express adenylate cyclase activity and possess membrane bound regulatory proteins that bind guanine nucleotides (G proteins). Stimulation of the cyclase by forskolin or addition of 8-bromo-cAMP augments progesterone secretion by cultured cytotrophoblasts at least in part, by promoting accumulation of components of the cholesterol side-chain cleavage system. This is the consequence of increased synthesis of the proteins participating in steroidogenesis as a result of the 8-bromo-cAMP-provoked increase in their respective mRNAs. We propose that progesterone synthesis by cytotrophoblasts is up-regulated by cyclic AMP, which acts to increase expression of genes encoding the steroidogenic machinery. Paracrine or autocrine factors may initiate this cascade by stimulating the cytotrophoblast adenylate cyclase. PMID- 2553956 TI - Characteristics of an antigonadotrophic GnRH-like protein in the ovaries of diverse mammals. AB - A GnRH-like protein, detected with a receptor assay for GnRH in rat ovarian membranes, is present in ovarian extracts of the rat, human, cow and ewe at levels much higher than immunoreactive GnRH. This protein has been partly purified and characterized from bovine ovaries. Highest levels of this protein are found in granulosa cells, but substantial activity is present in luteal, thecal and stromal tissue. No detectable GnRH-like activity is found in follicular fluid, or in ovarian venous or jugular plasma. The GnRH-like protein is structurally and immunologically different from GnRH, but it competitively and reversibly inhibits binding of GnRH to high-affinity sites in rat ovarian membranes. The GnRH-like protein markedly inhibits the action of both LH and FSH in cultures of rat luteal and granulosa cells, respectively, to a much greater extent than GnRH. Co-elution of receptor-binding activity and antigonadotrophic activity occurs within and between several chromatography procedures, but the protein has not been completely purified. The presence of this antigonadotrophic GnRH-like protein in the ovaries of diverse species raises the possibility that it may represent a novel paracrine regulator of ovarian function. PMID- 2553957 TI - Adaptations to pregnancy in the interactions between luteal oxytocin and the uterus in ruminants. PMID- 2553958 TI - Mechanism of action of gonadotrophin-releasing hormone on pituitary gonadotrophin secretion. PMID- 2553959 TI - Role of phosphoinositol metabolism and phospholipases C and A2/A1 in signal transduction in isolated rat adrenal cells. AB - Isolated rat adrenal glomerulosa cells were prelabelled with [3H]inositol and stimulated with 25 nM-angiotensin II in the presence of Li+. The resulting inositol monophosphates were separated using h.p.l.c. and 2 major peaks of radioactivity were detected. These showed the same characteristics as inositol 4 phosphate and inositol 1-phosphate and stimulation with angiotensin II increased activity 4-5 fold and 7-8-fold respectively. A minor peak with the characteristics of inositol 1:2 cyclic phosphate increased 1.5-fold after stimulation. No material corresponding to inositol 2-phosphate or inositol 5 phosphate was detected. The results establish the identify of the main inositol phosphate products in angiotensin II-stimulated rat glomerulosa cells. Analysis by h.p.l.c. has been similarly used to assess the inositol phosphates produced after ACTH1-39-stimulation of isolated rat adrenal fasiculata-reticularis cells. A low dose of ACTH1-39 (10(-12) M) stimulated production of small but significant amounts of both glycerophosphoinositol and inositol monophosphate. Using superfused isolated fasiculata-reticularis cells it was also found that ACTH1-39 (10(-9) M and 10(-12) M) rapidly increased efflux of 45Ca+ from 45Ca2+ prelabelled cells. It is concluded that although the results are consistent with a role for phospholipase C in the action of angiotensin II on adrenal glomerulosa cells, in the action of ACTH on adrenal fasciculata-reticularis cells a role for phospholipase A2/A1 is implicated. PMID- 2553960 TI - Landry-Guillain-Barre-Strohl syndrome in pregnancy. A report of two cases. AB - Landry-Guillain-Barre-Strohl syndrome (LGBS) in pregnancy is a rare event previously reported on only 29 times. Both cases reported on here were associated with cytomegalovirus infection. The fetus did not appear to be affected by the paralyzing agent in LGBS, although sequelae of the cytomegalovirus could occur later. Aggressive, long-term care is necessary to minimize maternal morbidity. PMID- 2553961 TI - Squamous atypia on cytology. The influence of age. AB - The optimal management of squamous atypia on cytology has not been determined. Colposcopy has been recommended because of the high false-negative rate of cytology. To evaluate colposcopy, 1,074 consecutive, nonpregnant women presenting with squamous atypia on cytology were examined prospectively with colposcopy and simultaneous repeat cytology. No specific treatment was given prior to evaluation. Biopsy and endocervical curettage were performed when clinically indicated. Two hundred two women (18.8%) had abnormal cervical or vaginal biopsies, 74 had human papillomavirus, and 128 had cervical intraepithelial neoplasia with or without associated papillomavirus infection. Invasive cancer was not detected. Repeat cytology suggested intraepithelial neoplasia in 60 women, 37 (61%) of whom had positive biopsy findings. Such biopsy findings were obtained in 89 of 306 women (29.1%) with repeat squamous atypia on cytology and in 76 of 708 women (10.8%) with repeat negative cytology. When the data were stratified in terms of age, 184/787 (19.6%) abnormal biopsies occurred in women less than or equal to 40 years of age and 18/287 (6.3%) in women greater than or equal to 41. Squamous atypia on cytology should not be ignored. However, women greater than or equal to 41 are at lower risk of having colposcopically detectable abnormalities and can be followed more conservatively. PMID- 2553962 TI - Evaluation of inflammatory atypia. A literature review. AB - The accepted procedure for evaluating women with inflammatory atypia on a Papanicolaou smear has changed over the years. Early studies reflected a general consensus that inflammatory atypia was normal and required no further evaluation. In the early 1970s several authors noted a higher-than-expected incidence of cervical intraepithelial neoplasia (CIN) and even cervical cancer in these patients. Further work in the late 70s and early 80s demonstrated that up to 79% of patients who had both initial and repeat atypical Papanicolaou smears had CIN or worse on biopsy. Those studies were flawed in their methodology. However, recent studies have been more sound methodologically. They clearly showed that 18.79%-56.00% of initially atypical smears are associated with CIN, human papillomavirus or cervical cancer. Furthermore, they demonstrated an alarmingly high number of false-negative repeat smears. Those studies pointed to a need for a more aggressive approach to the evaluation of inflammatory atypia seen on the initial smear. PMID- 2553963 TI - Histogenesis of cervical adenocarcinoma with reference to human papillomavirus-18 as a carcinogen. AB - Human papillomavirus (HPV) DNA is present in adenocarcinoma of the cervix as frequently as in squamous cell carcinoma of the cervix. Recent molecular biologic studies strongly suggested that HPV acts at least as a cocarcinogen in the female genital organs. Thus, a study of the presence of HPV DNA in adenocarcinoma in situ, endocervical glandular dysplasia and microglandular endocervical hyperplasia, three lesions suggested as possible precursor lesions of adenocarcinoma of the cervix, would clarify some aspects of the histogenesis of adenocarcinoma of the cervix. The presence of HPV-6, HPV-16 and HPV-18 in those precursor lesions was studied with highly sensitive in situ DNA hybridization. Fourteen of 21 cases of adenocarcinoma in situ (67%) contained HPV DNA; approximately the same proportion of HPV DNA was seen in invasive adenocarcinoma. Two of 36 cases of endocervical glandular dysplasia contained HPV DNA; both of them were lesions coexisting with adenocarcinoma in situ and thus might have been well-differentiated lesions of adenocarcinoma in situ. HPV DNA was not present in 16 cases of microglandular endocervical hyperplasia. Adenocarcinoma in situ may be the earliest event in HPV infection of the endocervical cells to lead to the development of adenocarcinoma of the cervix. PMID- 2553965 TI - Primary Sjogren's syndrome. Six characters in search of an author. PMID- 2553964 TI - Lower extremity peripheral neuropathy and ischemic ulcers associated with giant cell arteritis. AB - A patient with previously treated, temporal artery biopsy proven, giant cell arteritis (GCA) developed lower extremity ulcers and sensory neuropathy. Sural nerve biopsy showed epineural vessels with focal mild chronic inflammation. The lower extremity skin and nerve abnormalities improved after retreatment with prednisone. The rare manifestations of lower extremity skin and peripheral nervous system involvement associated with GCA are discussed. GCA must be considered in lower extremity ischemic processes of the skin and peripheral nervous system. PMID- 2553966 TI - Jaccoud's-type arthropathy: an association with malignancy. AB - We describe a patient who developed Jaccoud's type arthropathy as an initial manifestation of small cell anaplastic carcinoma of the lung. This is the first time this arthropathy has been reported as a musculoskeletal manifestation of malignancy. PMID- 2553967 TI - Evidence for the involvement of interleukin 1 in human osteoarthritic cartilage degradation: protective effect of NSAID. AB - Our study demonstrates for the first time the presence, in situ, of interleukin 1 (IL-1) in human osteoarthritic (OA) synovium as well as in cartilage. In vitro IL 1 was found to significantly increase the catabolism of proteoglycan macromolecules and to stimulate the metalloprotease activity in human OA cartilage explants. Tiaprofenic acid, a nonsteroidal antiinflammatory drug belonging to the propionic acid class, reduced both the proteoglycan catabolism (26 micrograms/ml, 6.3 +/- 1.4%; 2.6 micrograms/ml, 7.1 +/- 1.4%) and metalloprotease activity (26 micrograms/ml, 1.1 +/- 0.3 units/mg wet weight; 2.6 micrograms/ml, 1.1 +/- 0.2 units/mg wet weight) induced by IL-1 (10.2 +/- 2.3%; 1.7 +/- 0.4 units/mg wet weight). The effect of tiaprofenic acid was reversed by a pharmacological dose of either PGE2 or dibutyryl cAMP. PMID- 2553968 TI - Phospholipase A2 activity in human osteoarthritic cartilage. AB - Phospholipase A2 (PLA2) activity was measured in articular cartilage from normal and osteoarthritic (OA) human femoral heads. Protoglycanase and collagenase activity was determined in the same specimens using radiolabelled human proteoglycan subunit and type II collagen, respectively. Grossly normal and fibrillated OA cartilage samples showed a significant increase in PLA2 activity which was not found in osteophytic cartilage. PLA2 activity was found to be correlated with proteoglycanase but unrelated to collagenase activity. Tiaprofenic acid induced in vitro a concomitant increase in PLA2 and a decrease in proteoglycanase activity. PLA2 which may be activated by cytokines as well as mechanical factors is suggested as a key enzyme in chondrocyte metabolism regulation. Tiaprofenic acid is shown as a potential chondroprotective nonsteroidal anti-inflammatory drug. PMID- 2553969 TI - Bovine visna virus and the origin of HIV. PMID- 2553970 TI - Choanal atresia as a feature of ectrodactyly-ectodermal dysplasia-clefting (EEC) syndrome. AB - We report here a father and daughter with digital abnormalities, nasolacrimal duct obstruction, and variable alopecia. The father had a cleft lip and palate and the daughter had choanal atresia. We propose they both have the EEC syndrome and show the variable expressivity of this disorder. Choanal atresia has not been previously reported in this condition. PMID- 2553971 TI - Serologic and virologic analysis of type D simian retrovirus infection in a colony of Celebes black macaques. PMID- 2553972 TI - Electrogenic properties of the Na,K pump. PMID- 2553973 TI - Effect of mercurial compounds on net water transport and intramembrane particle aggregates in ADH-treated frog urinary bladder. AB - It has been suggested that during the oxytocin-induced hydrosmotic response, water crosses the luminal membrane of urinary bladder epithelium cells through membrane-spanning proteins. Although specific inhibitors of osmotic water transport have not been found, certain sulfhydryl reagents such as mercurial compounds may help to identify the proteins involved in this permeation process. We tested the effects of p-chloromercuribenzene sulfonate (PCMBS) and of fluorescein-mercuric acetate (FMA) on the net water flux, the microtubule and microfilament structures of the frog urinary bladder, and the distribution of intramembrane particle aggregates in the luminal membrane. We observed that: (i) 5 mM PCMBS at pH 5 and 0.5 mM FMA at pH 8 added to the mucosal bath at the maximum of the response to oxytocin partially inhibited the net water flux. Inhibition then increased progressively when the preparation was repeatedly or continuously stimulated, until it reached a maximal inhibition at 120 min. This inhibition was not reversed even when cystein was added in the mucosal bath. PCMBS and FMA effects were also observed when cyclic AMP (3',5' cyclic adenosine monophosphate) was used to increase water permeability, (ii) PCMBS mucosal pretreatment did not modify the basal water flux but potentiated the inhibitory effect of PCMBS or FMA on the hydrosmotic response to oxytocin. (iii) Microtubule and microfilament network, visualized in target cells by immunofluorescence, was not affected by PCMBS. (iv) The maximal PCMBS or FMA inhibition was not associated with a reduction of aggregate surface area in the apical membrane. The persistence of the intramembrane particle aggregates associated with the oxytocin induced hydrosmotic response during the net water flux inhibition by PCMBS, suggests that the PCMBS effect occurs possibly at the level of sulfhydryl groups of the water channel itself. PMID- 2553974 TI - Effect of K+ channels in the apical plasma membrane on epithelial secretion based on secondary active Cl- transport. AB - Models of epithelial salt secretion, involving secondary active transport of Cl- [9], locate the K+ conductance of the plasma membrane exclusively in the basolateral membrane, although there is considerable experimental evidence to show that many secretory epithelia do have a significant apical K+ conductance. We have used an equivalent circuit model to examine the effect of an apical K+ conductance on the composition and flow rate of the fluid secreted by an epithelium in which secretion is driven by the secondary active transport of Cl-. The parameters of the model were chosen to be similar to those measured in the dog tracheal mucosa when stimulated with adrenaline to secrete. We find that placing a K+ conductance in the apical membrane can actually enhance secretion provided the proportion of the total cell K+ conductance in the apical membrane is not greater than about 60%, the enabling effect on secretion being maximal when the proportion is around 10-20%. We also find that even when the entire cell K+ conductance is located in the apical membrane, the secreted fluid remains relatively Na+ rich. Analysis of the sensitivity of model behavior to the choice of values for the parameters shows that the effects of an apical K+ conductance are enhanced by increasing the ratio of the paracellular resistance to the transcellular resistance. PMID- 2553975 TI - Rabbit distal colon epithelium: I. Isolation and characterization of basolateral plasma membrane vesicles from surface and crypt cells. AB - A method has been developed for the simultaneous isolation of basolateral plasma membrane vesicles from surface and crypt cells of rabbit distal colon epithelium by sequential use of differential sedimentation, isopycnic centrifugation and Ficoll 400 barrier centrifugation. The protein yield was high (total 0.81 mg/g mucosa) and surface and crypt cell-derived basolateral membrane fractions have been purified 34- and 9-fold with respect to the homogenate. The pattern of marker enzyme enrichments revealed only minor contamination by subcellular organelles. Latency of ouabain-sensitive (Na+,K+)-ATPase activity prior and after trypsin treatment of membranes indicated a vesicle configuration of sealed right side-out: sealed inside-out: leaky of approximately 2:1:1. The presence of sealed vesicles was also evident from the osmotic sensitivity of the D-[1-14C] mannitol equilibrium space determined with either fraction. Although considerably different in protein profile, surface and crypt basolateral membranes were similar in cholesterol to phospholipid molar ratio and membrane fluidity as determined by steady-state fluorescence polarization. Stopped-flow light scattering experiments revealed a rather low water permeability of the membranes with a permeability coefficient of 6 microns/sec at 35 degrees C, which is one order of magnitude lower than reported for small intestinal plasma membranes. Both membrane fractions have been shown to effectively generate outward uphill potassium ion gradients, a process that is energized by ATP and inhibited by the membrane-permeant cardiac-glycoside digitoxin. These characteristics are consistent with the activity of a (Na+,K+) pump operating in inside-out vesicles. PMID- 2553976 TI - Polypeptide neurotoxins modify gating and apparent single-channel conductance of veratridine-activated sodium channels in planar lipid bilayers. AB - The effects of scorpion and sea anemone polypeptide toxins on partially purified veratridine (VER)-activated Na channels from rat brain were studied at the single channel level in planar lipid bilayers. The probability of the VER-activated channel being open (Po) increased with depolarization; Po was 0.5 at -40 to -50 mV. Saxitoxin (STX) blocked VER-activated channels with an apparent dissociation constant of about 1 nM at -45 mV. The apparent single-channel conductance was approximately 9 pS, similar to that seen in VER-activated Na channels from skeletal muscle transverse tubules. Addition of sea anemone or scorpion polypeptide toxins to VER-activated Na channels resulted in a 19% increase in apparent single-channel conductance and a hyperpolarizing shift in the Po vs. Vm relation such that the channels were more likely to be open at potentials less than 40 mV. These effects of the polypeptide toxins on the single-channel properties of VER-activated Na channels may account for the previously described potentiation of VER action by polypeptide toxins. PMID- 2553977 TI - Degradation of skeletal muscle plasma membrane proteins by calpain. AB - Observations described here provide the first demonstration that calpain (Ca2+ dependent cysteine protease) can degrade proteins of skeletal muscle plasma membranes. Frog muscle plasma membrane vesicles were incubated with calpain preparations and alterations of protein composition were revealed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Calpain II (activated by millimolar concentrations of Ca2+) was isolated from frog skeletal muscle, but the activity of calpain I (activated by micromolar concentrations of Ca2+) was lost during attempts at fractionation. Calpain I obtained from skeletal muscle and erythrocytes of rats was tested instead, and exerted effects similar to those of frog muscle calpain on the membrane proteins. All of the calpain preparations caused striking losses of a major membrane protein of molecular mass of approximately 97 kDa, designated band c, and diminution of a thinner band of approximately 200 kDa. There were concomitant increases in 83- and 77-kDa polypeptides. These effects were absolutely dependent on the presence of free Ca2+, and were completely blocked by calpastatin, a specific inhibitor of calpain action. Frog muscle calpain differed only in being relatively more active at 0 degree C than were the calpains from rat tissues. Experimental observations suggest that calpain acts at the cytoplasmic surface of the plasma membrane. PMID- 2553979 TI - In vivo assessment of the Z-DNA-forming potential of d(CA.GT)n and d(CG.GC)n sequences cloned into SV40 minichromosomes. AB - Alternating repeated d(CA.GT)n and d(CG.GC)n sequences constitute a significant proportion of the simple repeating elements found in eukaryotic genomic DNA. These sequences are known to form left-handed Z-DNA in vitro. In this paper, we have addressed the question of the in vivo determination of the Z-DNA-forming potential of such sequences in eukaryotic chromatin. For this purpose, we have investigated the ability of a d(CA.GT)30 sequence and a d(CG.GC)5 sequence to form left-handed Z-DNA when cloned into simian virus 40 (SV40) minichromosomes at two different positions: the TaqI site, which occurs in the intron of the T antigen gene, and the HpaII site, which is located in the late promoter region within the SV40 control region. Formation of Z-DNA at the inserted repeated sequences was analyzed through the change in DNA linkage associated with the B to Z transition. Our results indicate that regardless of: (1) the site of insertion (either TaqI or HpaII), (2) the precise moment of the viral lytic cycle (from 12 h to 48 h postinfection) and (3) the condition of incorporation of the SV40 recombinants to the host cells (either as minichromosomes or as naked DNA, relaxed or negatively supercoiled), neither the d(CA.GT)30 nor the d(CG.GC)5 sequence are stable in the left-handed Z-DNA conformation in the SV40 minichromosome. The biological relevance of these results is discussed. PMID- 2553980 TI - Regulation of IS1 transposition by the insA gene product. AB - The IS1 element contains two adjacent genes called insA and insB, both required for IS1 transposition and IS1-mediated plasmid cointegration. These two genes are transcribed polycistronically from the promoter in the left terminal inverted repeat of IS1 (insL). We constructed overexpression systems of these genes with the tac promoter, which are regulated by an exogenous inducer, isopropyl-beta-D thiogalactopyranoside (IPTG). Then we have examined, under various conditions of induction with IPTG, how overexpression of these genes affects IS1 transposition, using an assay based on plasmid cointegration. When the insA and insB genes were organized identically to the wild-type IS1 genes and simultaneously expressed using low concentrations of IPTG, activity of a mutant IS1 in cis was restored, but not in trans. Higher IPTG concentrations resulted in lower transposition activity. Expression in trans of insA and insB results in a 50 to 100-fold reduction of the frequency of cointegration mediated by wild-type IS1. Such a reduction is also observed when only the insA gene is overexpressed in trans. Overexpression of either mutant insA or insB does not affect the cointegration event. Tests with the insA-lacZ fusion gene showed that the InsA product inhibits the expression of IS1 genes directed by its own promoter in insL. These results suggest that the InsA product regulates IS1 transposition by inhibiting expression of IS1 transposition genes in addition to acting as part of a transposase complex. PMID- 2553978 TI - Role of kinases in insulin stimulation of glucose transport. PMID- 2553981 TI - Mapping of insertion elements IS1, IS2 and IS3 on the Escherichia coli K-12 chromosome. Role of the insertion elements in formation of Hfrs and F' factors and in rearrangement of bacterial chromosomes. AB - The chromosome of an Escherichia coli K-12 strain W3110 contains seven copies of insertion element IS1, 12 copies of IS2 and six copies of IS3. We determined the approximate locations of six copies of IS1 (named is1A to is1F), ten copies of IS2 (named is2A to is2J), and five copies of IS3 (named is3A to is3E) on the W3110 chromosome by plaque hybridization using the "mini-set" of the lambda phage library that includes 476 clones carrying chromosomal segments that cover the W3110 chromosome almost entirely. Cleavage maps of the W3110 chromosome and cleavage analysis of phage DNAs carrying insertion elements allowed us to assign more precise locations to most of the insertion elements and to determine their orientations. Insertion elements were distributed randomly along the W3110 chromosome in one or other orientation. Several of these were located at the same positions on the chromosome of another E. coli K-12 strain, JE5519, and they were assumed to be the original complement of insertion elements in E. coli K-12 wild type. Locations and orientations of such insertion elements were correlated well with Hfr points of origin and with crossover points for excision of some F' factors derived from several Hfrs. Insertion elements may be involved also in rearrangement of bacterial chromosomes. PMID- 2553982 TI - Preparation of heavy-atom derivatives using site-directed mutagenesis. Introduction of cysteine residues into gamma delta resolvase. AB - The ability to determine protein structures by X-ray crystallography is often thwarted by the difficulty of finding isomorphous heavy-atom derivatives. The crystal structure of the site-specific recombinase, resolvase, has been difficult to determine for this reason. We have overcome this problem by introducing 13 single cysteine substitutions into the resolvase catalytic domain using oligonucleotide mutagenesis. The mutant proteins were screened for their ability to crystallize into the orthorhombic form and bind mercury ions isomorphously. Two mutant proteins provided excellent heavy-atom derivatives. This approach should be of general use and particularly helpful in cases where traditional methods have failed to produce a derivative. PMID- 2553984 TI - Complement genes C1r and C1s feature an intronless serine protease domain closely related to haptoglobin. AB - The exon-intron structure of the human complement C1s gene displays a striking similarity with that of the gene encoding haptoglobin, a peculiar transport protein distantly related to the serine proteases. While the protease regions of the serine zymogens are typically encoded by multiple exons, the protease domains of C1s and of its genetically linked and functionally interacting homolog C1r are encoded as intronless domains, not unlike a region of haptoglobin, which in fact is devoid of proteolytic activity. The close similarity of the C1s gene with haptoglobin includes the precise conservation of exon-intron junctions and it extends to upstream exons encoding the short repeats typical of several complement components, but found also in other functionally unrelated proteins. Additional evidence of the common ancestry of C1r, C1s and haptoglobin is the presence, within the protease domain, of a set of sequence markers that distinguish these three proteins from all known serine proteases. The finding of vertebrate serine protease genes with an uninterrupted protease-encoding exon supports the definition of a novel evolutionary branch of this gene family and rules out the hypothesis that regards this unusual exon as an irrelevant byproduct of the extravagant functional divergence of haptoglobin. PMID- 2553983 TI - Crystal structure of the p-hydroxybenzoate hydroxylase-substrate complex refined at 1.9 A resolution. Analysis of the enzyme-substrate and enzyme-product complexes. AB - Using synchrotron radiation, the X-ray diffraction intensities of crystals of p hydroxy-benzoate hydroxylase, complexed with the substrate p-hydroxybenzoate, were measured to a resolution of 1.9 A. Restrained least-squares refinement alternated with rebuilding in electron density maps yielded an atom model of the enzyme-substrate complex with a crystallographic R-factor of 15.6% for 31,148 reflections between 6.0 and 1.9 A. A total of 330 solvent molecules was located. In the final model, only three residues have deviating phi-psi angle combinations. One of them, the active site residue Arg44, has a well-defined electron density and may be strained to adopt this conformation for efficient catalysis. The mode of binding of FAD is distinctly different for the different components of the coenzyme. The adenine ring is engaged in three water-mediated hydrogen bonds with the protein, while making only one direct hydrogen bond with the enzyme. The pyrophosphate moiety makes five water-mediated versus three direct hydrogen bonds. The ribityl and ribose moieties make only direct hydrogen bonds, in all cases, except one, with side-chain atoms. The isoalloxazine ring also makes only direct hydrogen bonds, but virtually only with main-chain atoms. The conformation of FAD in p-hydroxybenzoate hydroxylase is strikingly similar to that in glutathione reductase, while the riboflavin-binding parts of these two enzymes have no structural similarity at all. The refined 1.9 A structure of the p-hydroxybenzoate hydroxylase-substrate complex was the basis of further refinement of the 2.3 A structure of the enzyme-product complex. The result was a final R-factor of 16.7% for 14,339 reflections between 6.0 and 2.3 A and an improved geometry. Comparison between the complexes indicated only small differences in the active site region, where the product molecule is rotated by 14 degrees compared with the substrate in the enzyme-substrate complex. During the refinements of the enzyme-substrate and enzyme-product complexes, the flavin ring was allowed to bend or twist by imposing planarity restraints on the benzene and pyrimidine ring, but not on the flavin ring as a whole. The observed angle between the benzene ring and the pyrimidine ring was 10 degrees for the enzyme substrate complex and 19 degrees for the enzyme-product complex. Because of the high temperature factors of the flavin ring in the enzyme-product complex, the latter value should be treated with caution. Six out of eight peptide residues near the flavin ring are oriented with their nitrogen atom pointing towards the ring.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2553985 TI - Densely packed beta-structure at the protein-lipid interface of porin is revealed by high-resolution cryo-electron microscopy. AB - Porin is an integral membrane protein that forms channels across the outer membrane of Escherichia coli. Electron microscopic studies of negatively stained two-dimensional porin crystals have shown three stain accumulations per porin trimer, revealing the locations of pores spanning the membrane. In this study, reconstituted porin lattices embedded in glucose were investigated using the low dose technique on a cryo-electron microscope equipped with a helium-cooled superconducting objective lens. The specimen temperature was maintained at 5 K to yield an improved microscopic and specimen stability. Under these conditions, we obtained for the first time electron diffraction patterns from porin lattices to a resolution of 3.2 A and images showing optical diffraction up to a resolution of 4.9 A. Applying correlation averaging techniques to the digitized micrographs, we were able to reconstruct projected images of the porin trimer to a resolution of up to 3.5 A. In the final projection maps, amplitudes from electron diffraction and phases from these images were combined. The predominant feature is a high-density narrow band (about 6 A in thickness) that delineates the outer perimeter of the trimer. Since the molecule consists of almost exclusively beta sheet structure, as revealed by spectroscopic data, we conclude that this band is a cylindrical beta-pleated sheet crossing the membrane nearly perpendicularly to its plane. Another intriguing finding is a low-density area (about 70 A2) situated in the centre of the trimer. PMID- 2553986 TI - Ca-mediated action potentials in ventricular myocardium. Variation by various stimulation patterns and by K-channel blockers. AB - In rabbit papillary muscle, Ca-mediated action potentials (AP's) and maximum rate of rise (Vmax) were measured at 18 mM Ko. At steady state stimulation Vmax, overshoot and diastolic membrane potential were enhanced with increasing cycle length (CL) (from 8.7 to 14.1 V/s, from 25.1 to 34.1 mV, from -34.5 to -49.6 mV, respectively, between the CL of 1 and 60 s). AP-duration was only slightly prolonged (from 109 to 138 ms at 50% repolarization). 4-aminopyridine (5 mM) caused no or minor changes in the magnitude and CL-dependence of Vmax and overshoot. The AP-duration, however, was strongly prolonged due to a slower repolarization of the plateau phase with increasing CL. CsCl (20 mM) caused similar changes. Ca-mediated AP's elicited after a period of rest were found to be qualitatively similar to those which were obtained at increased steady state CL. Vmax of premature Ca-mediated AP's increased exponentially with the interval of stimulation, t, and was accurately described by: Vmax = A(1 - exp - (t T)/tau). The K-channel blockers had no significant effect on the absolute refractory period, T, but increased the time constant, tau, from 123 to 166 ms. The restitution of AP-duration nicely matched this time course. The voltage dependence of membrane responsiveness showed a sigmoid shape, and was accurately described by: Vmax = A(1 + exp(V-Vs)/s)-1. The K-channel blockers increased s from 2.2 to 3.6 mV and shifted Vs from -26.8 to -22.1 mV.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553987 TI - Reduction of calcium channel antagonist binding sites by oxygen free radicals in rat heart. AB - In view of the importance of Ca2+-channels in controlling the entry of Ca2+ into the myocardium, this study was undertaken to examine the effects of oxygen free radicals on the binding of Ca2+-channel antagonists in rat heart by employing [3H]-nitrendipine as a ligand. Isolated heart membranes were incubated with xanthine + xanthine oxidase (a superoxide anion radicals generating system), hydrogen peroxide (an activated species of oxygen), or hydrogen peroxide + Fe2+ (a hydroxyl radicals generating system). The assay of the [3H]-nitrendipine binding activity revealed that the maximal number of binding sites (Bmax) were reduced in a time-dependent manner by superoxide radicals without any changes in the binding constant (Kd); a significant reduction of Bmax was seen after incubating membranes with xanthine + xanthine oxidase for a 10-min-period. Superoxide dismutase showed a protective effect on the superoxide radicals induced reduction in Bmax. Both hydrogen peroxide and hydroxyl radicals also depressed the Bmax for [3H]-nitrendipine binding without any significant change in Kd; catalase and mannitol showed protective effects on hydrogen peroxide or hydroxyl radicals induced depression in Bmax, respectively. These results indicate that oxygen free radicals may reduce the number of Ca2+-channels in the cell membrane and this change may contribute towards decreasing the voltage dependent Ca2+ influx in the cardiac cell. PMID- 2553989 TI - Papillomavirus in anogenital cancer: the dilemma of epidemiologic approaches. PMID- 2553988 TI - Scrambled duplications in the feline leukemia virus gag gene: a putative pattern for molecular evolution. AB - The present study is a detailed computer-assisted analysis of the feline leukemia virus gag gene nucleotide sequence together with its flanking sequences (ST-FeLV GAG) that is compared with the aligned sectors of the Moloney strain of murine leukemia virus (Mo-MuLV GAG) and of three strains of feline sarcoma virus. It shows that perfectly matched repeated oligomers up to 13 nucleotides long are overrepresented and scattered throughout both ST-FeLV GAG and Mo-MuLV GAG, in noncoding and coding sectors, with no stringent correlation to codon usage in ST FeLV gPr80gag. Many repeated oligomers share a core consensus that is intriguingly part of the inverted repeat at the termini of the long terminal repeat. Local scrambled repetitions of nucleotide subsequences have been found; they suggest a model of molecular evolution by slippage-like mechanisms. Thus, viral genomes could be subject to the same evolutionary mechanisms that are now known to be operating extensively in eukaryotic genomes. The data are discussed in light of putative patterns of molecular evolution. PMID- 2553990 TI - Unknotting the complexities of multidrug resistance: the involvement of DNA topoisomerases in drug action and resistance. PMID- 2553991 TI - Antibodies against the human papillomavirus type 16 early proteins in human sera: correlation of anti-E7 reactivity with cervical cancer. AB - By Western blot technique, 519 samples of human sera were tested for the presence of antibodies to the human papillomavirus (HPV) type 16 proteins E4 and E7 that had been expressed in Escherichia coli as fusion proteins. Sera were obtained from patients attending the University hospitals for reasons unrelated to HPV infections (controls), from patients with HPV-associated lesions, as well as from patients suffering from cervical cancer. Within the control population, 18.1% of them had antibodies that reacted with the E4 protein, and 3.9% of them had antibodies that reacted with the E7 protein. No sex-specific difference in the antibody prevalence was observed. The highest proportion of anti-E4 antibody positive individuals (40.7%) was observed in the age group between 11 and 20 years. The frequency of anti-E4-positive sera was threefold higher in patients with HPV-associated genital lesions than that in age-matched controls. Antibodies against the HPV16 E7 protein were found 14 times more frequently in patients with cervical cancer, compared with age- and sex-matched controls (P less than .00001). From these data, we concluded that anti-E4 antibodies may be correlated with virus replication and that anti-E7 antibodies may represent a marker for cervical cancer development. PMID- 2553992 TI - Nonproductive rearrangement of DNA topoisomerase I and II genes: correlation with resistance to topoisomerase inhibitors. AB - Topoisomerase inhibitors comprise an important group of agents that is used in cancer treatment. Because the development of resistance to cancer chemotherapeutic agents represents a major limitation of cancer chemotherapy, we investigated the mechanism of resistance by murine P388 leukemia to camptothecin (topoisomerase I inhibitor) or amsacrine (topoisomerase II inhibitor). The resistant cells contained reduced levels of topoisomerase activity and messenger RNA. The topoisomerase gene of these cells was rearranged (only in one allele) and hypermethylated. These topoisomerase gene alterations probably resulted in reduced transcription and, thus, enzyme production, which was correlated with resistance to the topoisomerase inhibitor. PMID- 2553993 TI - Randomized phase II-III trial of combination beta and gamma interferons and etoposide and cisplatin in inoperable non-small cell cancer of the lung. AB - We observed major responses in two patients with adenocarcinoma of the lung who had received a combination of interferon (IFN)-beta and IFN-gamma, immediately followed by chemotherapy. To verify these observations, we initiated a prospective randomized phase II-III trial of etoposide and cisplatin, with or without IFN-beta and IFN-gamma, in patients with inoperable non-small cell lung cancer. Thirty-seven patients were randomized to receive either two cycles of chemotherapy or 6 weeks of IFN-beta plus IFN-gamma followed by two cycles of chemotherapy. Chemotherapy consisted of 60 mg of cisplatin/m2 on day 1 and 120 mg of etoposide/m2 on days 4, 6, and 8, repeated every 21 days. Patients who were randomized to the IFN plus chemotherapy arm received 200 micrograms of IFN-gamma and 30 x 10(6) U of IFN-beta three times per week for 6 weeks, followed by two cycles of chemotherapy. Three of 18 (17%) eligible patients in the chemotherapy arm and two of 18 (11%) patients in the combination arm had partial responses. All responses occurred while patients were receiving chemotherapy. Median survival was 190 days for the chemotherapy arm and 246 days in the combined modality arm, as estimated from Kaplan-Meier curves (P = .35). We observed no significant difference in subjective toxic effects between the two arms. We observed more hematologic toxicity during chemotherapy on the combined modality arm (P = .02). We conclude that pretreatment with IFN-beta and IFN-gamma does not enhance the efficacy of etoposide and cisplatin in this disease. Although the combined modality arm is relatively well tolerated, it does result in more chemotherapy-associated toxic effects. This study also exemplifies a hybrid phase II-III trial design, which is useful in allowing phase II results to be quickly incorporated into a phase III trial. PMID- 2553994 TI - Glutamate receptor agonists increase the expression of Fos, Fra, and AP-1 DNA binding activity in the mammalian brain. AB - Administration of convulsant doses of Metrazole (pentylenetetrazol) and picrotoxin, as well as maximal electroshock, results in a rapid but transient increase in c-fos mRNA in mouse brain. Elevation of c-fos mRNA is followed by the accumulation and subsequent disappearance of Fos, the protein encoded by c-fos. In addition, immunoblots reveal the induction of two additional proteins that are antigenically related to Fos (Fra, Fos-related antigens). Fos and the various Fra appear and disappear in a staggered manner over an 8 hour period, such that at longer times after stimulation the brain contains no Fos but relatively large amounts of Fra (the latter being designated here by their apparent molecular weights, Fra-46K and Fra-35K). Previous studies have established that Fos, as well as several Fra, contribute to transcription factor AP-1 nucleoprotein complexes along with Jun, the product of the jun proto-oncogene. The appearance in brain of Fos and Fra coincides with a protracted increase in total AP-1 DNA binding activity, indicating that all the Fos-like proteins can participate in AP 1 complexes. Furthermore, the molecular composition of these complexes alters with time after stimulation. The induction of c-fos by Metrazole is blocked or attenuated by known anticonvulsants such as diazepam and valproate as well as the N-methyl-D-aspartate (NMDA) receptor antagonists, 2-amino-5-phosphonovaleric acid (APV) and MK-801. This suggests that fos induction might involve stimulation of a glutamate receptor. This conclusion was strengthened by the observations that two glutamate receptor agonists, kainic acid and NMDA, induced c-fos expression.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2553995 TI - Involvement of alpha-adrenoreceptors in the regulation of omasal cyclic myoelectrical activity in sheep. AB - In five conscious adult ewes at rest, chronically implanted with electrodes in the musculature of the omasal wall, intravenous (i.v.) infusion for 30 min of alpha 1- or alpha 2-adrenergic receptor blockers, prazosin (20 micrograms/kg/min) and yohimbine (30 micrograms/kg/min), respectively, had no significant effects on omasal myoelectrical activity. The i.v. administration for 15 min of alpha 1- or alpha 2-agonists phenylephrine (4 micrograms/kg/min) or naphazoline (2.5 micrograms/kg/min), respectively, increased the frequency and the amplitude of groups of myoelectrical discharges of omasum, as well as the duration of its activity. Pretreatment of animals with prazosin blocked the responses to phenylephrine. Yohimbine prevented the effects of naphazoline dose-dependently. It is suggested that both alpha 1- and alpha 2-adrenoreceptors are involved in regulation of the sheep's omasal notility. This regulation did not seem to be a simple consequence of the changes in the reticular motility. PMID- 2553996 TI - Preliminary pharmacokinetics of 2',3'-dideoxycytidine in neonatal goats. PMID- 2553997 TI - Morphology of mineral deposits on encrusted urinary catheters investigated by scanning electron microscopy. AB - Struvite and hydroxyapatite were precipitated from artificial urine onto the surfaces of catheter materials by the controlled addition of urease. They were precipitated both together and separately (by omitting components of the artificial urine), and with and without the inclusion of albumin (which was intended to mimic the proteinaceous debris found in infected urine). Precipitates were identified by X-ray powder diffraction and the artificially encrusted surfaces examined by scanning electron microscopy. In the presence of protein, hydroxyapatite was precipitated as a poorly crystalline form which aggregated to form a crust. Struvite crystals could be easily identified under the scanning electron microscope by their relatively large size and characteristic appearance. Fifteen encrusted catheters from patients were also examined by scanning electron microscopy, and a further six using X-ray microanalysis. Their appearance was very similar to that of the materials encrusted in vitro. Encrustation involves the formation of hydroxyapatite and the growth of struvite crystals, intimately associated with bacteria. PMID- 2553998 TI - Alpha-adrenoceptor function in isolated penile circumflex veins from potent and impotent men. AB - Alpha-adrenoceptor functions were investigated in isolated human penile circumflex veins from six potent and four impotent men. Contractions elicited by noradrenaline and phenylephrine were inhibited by prazosin, yohimbine, phentolamine and papaverine. No differences were found between vessels from potent and impotent men. The results suggest that alpha-adrenoceptors in penile circumflex veins are of both alpha 1- and alpha 2-type, and that no changes in alpha-adrenoceptor function occur in impotence associated with an increased penile venous outflow. PMID- 2553999 TI - A review of marble spleen disease of ring-necked pheasants. AB - Marble spleen disease (MSD) is a contagious disease of captive-reared ring-necked pheasants (Phasianus colchicus) characterized by variable mortality and high morbidity. The etiologic agent is a type II avian adenovirus closely related to hemorrhagic enteritis virus of turkeys and splenomegaly virus of chickens. Gross lesions of MSD consist of enlargement and mottling of the spleen and pulmonary edema. Microscopic alterations are most pronounced in the spleen and are characterized by reticuloendothelial (RE) cell hyperplasia and lymphoid necrosis. Intranuclear viral inclusions occur within hyperplastic RE cells. Serologic detection of MSD is routinely accomplished using an agar gel precipitin test. Oral vaccination with hemorrhagic enteritis virus or avirulent strains of MSD virus is effective in preventing MSD. There is no specific treatment available for MSD but supportive care, strict sanitation and good management practices are helpful in reducing mortalities and limiting the spread of an epizootic. PMID- 2554000 TI - Serologic survey for infectious canine hepatitis virus in grizzly bears (Ursus arctos) from Alaska, 1973 to 1987. AB - Serum antibody prevalence of infectious canine hepatitis virus was 12% (90 of 725) for grizzly bears (Ursus arctos) from Alaska (USA) during the period 1973 to 1987. Prevalence was highest on Kodiak Island at 29% (37 of 127). Prevalence of exposure at individual collection areas did not change significantly over time. There were no significant sex-specific differences in prevalence. Prevalence was directly related to age, but it was 0% for bears less than 2-yr-old. Young bears which are exposed to the virus may develop clinical disease and die as a result of the infection. This disease may be a factor affecting grizzly bear population dynamics. PMID- 2554001 TI - Prevalence and transmission of pseudorabies virus in an isolated population of feral swine. AB - Six hundred sixty-one feral swine (Sus scrofa) from Ossabaw Island, Georgia (USA) were captured, bled, and their sera tested for pseudorabies virus (PRV) antibody during a 6 yr period. Prevalence of seroconversion in females was somewhat higher than in males (10% versus 7%), but the difference was not statistically significant. Adults had a significantly higher prevalence than juveniles (29% versus 1%). An important finding in this study was that seroconversion occurred primarily in the adult feral swine. PMID- 2554002 TI - Herpes-like viral dermatitis in a beluga whale (Delphinapterus leucas). AB - Approximately 3.5 mo following its capture, a beluga whale (Delphinapterus leucas) developed focal pale gray skin lesions. These lesions persisted for at least 8 mo. A biopsy from one of these sites revealed epithelial intranuclear inclusions. Herpes-like viral particles were seen by transmission electron microscopy. The eventual regression of skin lesions and lack of other clinical signs suggests the virus was only mildly pathogenic in this animal. PMID- 2554003 TI - Influence of beta-blockade on ejection fraction, ANP, and cGMP in patients after myocardial infarction. PMID- 2554004 TI - [Results of conventional radiotherapy in malignant gliomas:]. AB - Results of radiotherapy for 164 supratentorial malignant gliomas were analyzed based on recent histological subclassification into glioblastoma multiforme (GBM) and anaplastic astrocytoma (AA). Patients with AA had a better prognosis than those with GBM. The 5-year survival rate was 1% for GBM and 16% for AA. The size of radiation field did not influence survival significantly, and whole brain radiation appeared of no benefit. Survival time prolonged with an increase of radiation dose between 45 Gy and 72 Gy. The optimal radiotherapy for the disease is discussed. PMID- 2554005 TI - [Three-dimensional analysis of regrowth pattern in recurrent supratentorial glioblastoma multiforme and anaplastic astrocytoma with special reference to prognosis]. AB - To determine effects of surgical treatments on the quality and duration of survival following tumor recurrence and to analyze prognostic factors that may affect the outcome, we have reviewed our experience with reoperations for 43 patients having recurrence with malignant histologic findings among 188 cases of supratentorial glioblastoma multiforme (GM), anaplastic astrocytoma (AA) and low grade astrocytoma (LGA) (27 of 77 patients with GM, and 16 of 42 patients with AA) since introduction of computed tomography (CT) in May, 1976. Using CT and/or more recent magnetic resonance imaging (MRI), the regrowth patterns of tumor recurrence were examined with aspect of local extension, non-contiguous or remote metastatic invasion, and tumor cell dissemination through cerebrospinal fluid. MRI can provide more significant informations on the selection for reoperation in terms of recognition of the developmental features. In most of the patients who underwent reoperations at recurrence, surgically accessible components of the lesion were shown to cause neurological deficits as a result of compression rather than infiltration of functionally important areas. Reoperations thus might be achieved with acceptable mortality and morbidity. In contrast, patients with tumors in inaccessible areas of the central nervous system received additional chemotherapy and/or interferon administration. Indications for re-irradiation of the recurrence is also discussed. PMID- 2554006 TI - [Regrowth pattern of glioblastoma multiforme after radiotherapy]. AB - We investigated the correlation between the technique of radiation therapy used and the regrowth pattern of recurrent glioblastoma multiformes of 48 patients who received initial radiation therapy in our clinic from April, 1974 to March, 1988. Three different techniques have been applied to patients with brain tumors: whole brain irradiation, generous local irradiation (the parallel opposing technique or three field technique, in which the treatment field fully covers the peritumoral low density area on CT images), and restricted local irradiation (the rotation technique, in which the treatment field is restricted to within about 2 cm of the tumor margin on CT images). Radiation dose of the treatment field was over 45 Gy in every case. The regrowth pattern was defined as being one of the followings: inside of the treatment field, outside of the treatment field (this includes the boundary zone between the treatment and the non-treatment field), and remote from the treatment field-which mostly due to the tumor spreading through ventricular systems. In all 7 cases that received whole brain irradiation, tumor recurred inside of the treatment field. Two of these 7 cases showed remote recurrence at the same time. Mean duration time to recurrence was 36.3 weeks. In 27 (90%) of 30 cases that received the generous local irradiation, tumor recurred within the treatment field. Only one showed outside recurrence, and two other cases showed remote recurrence. The mean duration time to recurrence in this group was 32.2 weeks. Of 11 cases that received restricted local irradiation, 3 cases (27%) showed recurrence outside of the treatment field, one showed recurrence in a remote area. The mean duration time to recurrence in this group was 38.3 weeks, but was not significantly longer than that of other two groups. These results indicate that restricted local irradiation sometimes fails to cover the tumor invaded area, and that the results of treatment using generous local irradiation are almost the same as those using whole brain irradiation. This suggests that generous local irradiation is advantageous to patients with brain tumors since the incidence of subacute side effects such as mental deterioration is much lower in local irradiation as compared with whole brain irradiation. PMID- 2554007 TI - [Autopsy cases of glioblastoma multiforme: treatment results of high-dose fractionated radiation therapy and CT scan findings]. AB - Six autopsy cases of glioblastoma multiforme in cerebral hemisphere were examined by large histological preparations. They were treated by surgery and high-dose fractionated radiation therapy (5 Gy twice weekly). Their morphological changes were compared to the last CT and radiation field and total doses. Four out of six cases showed small residual tumor. One case showed extensive necrosis of the tumor and brain. The other case exhibited no tumor tissue at all. Spongy degeneration of the white matter associated with astrocytosis and macrophage infiltration extended sometimes beyond the local irradiation field. These white matter changes were easily occurred in the previous peritumoral edema where tumor cell infiltration was frequently observed. Residual tumor cells consisted of small anaplastic cells, which might be radioresistant and recur. Enhancement effect of CT scan showed tumor tissue and radiation necrosis with vascular proliferation. PMID- 2554008 TI - [A clinicopathological study on colonic carcinomas arising from right side colon]. AB - This paper discusses the clinicopathological characteristics of colonic carcinomas arising from right side colon. Out of 214 cases of resected colonic carcinomas during the past 13 years, 36 cases of a right side colon carcinoma (in the cecum, in the ascending colon, or in the hepatic flexure) were encountered. Several characteristics have been associated with this type of cancer. They are as follows: 1) in half of these cases an abdominal pain but no anal bleeding, as has been noted in carcinomas of the sigmoid or rectum; 2) in histological type, poorly differentiated adenocarcinomas or mucinous carcinomas in 30.5% of these 36 cases, and associated with a far distant nodal involvement (n3, n4) in advanced carcinomas; and 3) although there was no difference in the 5-year survival rate between a right side colon carcinoma or a colon carcinoma at another site, the prognosis for those given a curative resection for a right side colon carcinoma was poor--only 22% due to hepatic metastases, indicating that colonic carcinomas of the right side has a poor prognosis due to the subsequent risk of highly potential malignancies. PMID- 2554009 TI - [Investigation of two cases of a clinically unknown large cell carcinoma of the lung involved with severe stenosis of the bilateral main bronchi]. AB - Reported are two rare cases of a clinically unknown large cell carcinoma of the lung, associated with severe stenosis of the bilateral main bronchi. Case 1: A 61 year-old man was admitted to our hospital because of chief complaints that included a cough, hemosputum, and dyspnea. Based on the bronchoscopic findings that revealed severe stenosis of the bilateral main bronchi and a specimen that was biopsied from tracheal spur, a diagnosis of large cell carcinoma of the lung was made. Although anticancer chemotherapy and irradiation therapy produced a slight improvement in the patient's symptoms, at the 10th hospital week the patient died of suffocation. Case 2: A 77-year woman was admitted to hospital with chief complaints that included a cough and hemosputum. A bronchoscopy revealed severe stenosis of the bilateral main bronchi. After the 4th hospital day the patient complained of increasing dyspnea. At the 9th hospital day the patient died of suffocation. An autopsy results revealed a large cell carcinoma (giant type) of the lung. PMID- 2554010 TI - [A hepatocellular carcinoma revealed by paraplegia caused by a vertebral metastasis]. AB - Described is a 57-year-old male with a complaint of a gait disturbance. On admission to hospital, he was suffering from almost complete paraplegia. Roentgenograms of the spine showed extensive destruction in the body of the second thoracic vertebra. Magnetic resonance computed tomography revealed a vertebral tumor that was compressing the vertebral cord. Thus, a laminectomy of the 1st to 3rd thoracic vertebra was performed to relieve the compression. Histological examination of the tumor showed it to be a metastatic vertebral tumor from a hepatocellular carcinoma (HCC). Subsequently, by abdominal computed tomography and an examination of the AFP serum level the existence of the HCC was confirmed. This is a very rare case of an HCC that was revealed by paraplegia caused by bone metastasis. PMID- 2554011 TI - [Breast carcinoma with features of osteosarcoma--a light and electron microscopic and immunohistochemical study]. AB - Reported is the case history of a 58-year-old woman with a lump in the left breast. The tumor was composed of two nodules, sharing a common portion. Grossly, one was an irregular hard tumor that was grayish in color, and the other was a well circumscribed cystic tumor that showed necrosis and a hemorrhage that had filled it with a reddish soft mass. Light microscopically, combined features of a scirrhous carcinoma and of an osteosarcoma were observed in each nodule, but the common portion was consistent with a metaplastic carcinoma, with cancer cells and sarcoma-like cells closely mingled. An ultrastructural study showed that the sarcoma-like cells were composed of polymorphic cells that resembled osteoblasts, myofibroblasts, osteoclasts, histiocytes, and undifferentiated tumor cells. Immunohistochemically, vimentin and alpha 1-antitrypsin in the sarcoma-like cells were positive, suggesting these cells were of a mesenchymal rather than of an epithelial origin. PMID- 2554013 TI - [Sezary syndrome associated with hepatocellular carcinoma and monoclonal gammopathy--a case report]. AB - We treated 54-year-old Japanese man with a large cell type of Sezary syndrome. He had generalized erythrodermia, superficial lymphadenopathy, atypical lymphocytes in the peripheral blood, anti-HTLV-I antibody negativity and chromosomal abnormality. The patient was a hepatitis B virus carrier, and was complicated with hepatocellular carcinoma and monoclonal gammopathy of IgG, lambda type. Sezary syndrome is a T cell malignancy, the clinical course of which is relatively mild and chronic; accordingly, this case showed no crisis under chemotherapy. However, the patient died due to rapid growth of the hepatoma. Although case reports of Sezary syndrome complicated with other malignancies are very few, the occurrence of malignancies is possible because of decreased immunological function in the patients. In this case, hepatitis B virus might participate in the hepatic oncogenesis under dysfunction of helper/inducer cells. In addition, the complication of monoclonal gammopathy was also interesting from the standpoint of the helper function of Sezary cells. PMID- 2554012 TI - [Germ-line configuration of the immunoglobulin heavy chain gene in a case of B cell precursor acute lymphoblastic leukemia]. AB - A 20-year-old man was admitted to our hospital because of fever and knee joint pain on March 20, 1986. Physical examination revealed generalized lymphadenopathy and hepatomegaly. White blood cell count was 32,800 microliters with 74.4% blast cells. Bone marrow was hypercellular with 93.6% blast cells. Blast cells were weakly positive for acid phosphatase and PAS stainings but were negative for peroxidase, sudan black B and esterase stainings. Cell surface marker analysis of blast cells disclosed that they were positive for anti-HLA-DR, CD19, CD24, CD33 and CD38, but were negative for CD10 and CD20. Cytoplasmic immunoglobulin of blast cells was negative and TdT activity by immunofluorescent method was positive. Chromosomal analysis of bone marrow samples revealed normal karyotype. Therefore, this case was diagnosed as having acute lymphoblastic leukemia (L2) and achieved complete remission with LVP therapy consisting of 1-asparaginase, vincristine and prednisolone. Gene analysis of blast cells disclosed germ-line configuration of both the immunoglobulin heavy chain gene and T cell receptor beta chain gene. We speculated that the phenotype of leukemic cells might precede the genotype in some cases of acute leukemia. PMID- 2554014 TI - [Malignant lymphoma which contained EBV DNA and lacked EBNA2 antibody]. AB - A case of non-Hodgkin malignant lymphoma which contained EBV DNA and EBNA was reported. The serum of this patient showed high titer of VCA, EA, and EBNA1 antibody but lacked EBNA2 antibody. The relation with EBV originated malignancy and defect of EBV specific antibody suggests a process of oncogenicity of this virus. PMID- 2554015 TI - [Progress in methods of studying renal tubular transport. Determination of intracellular ion activities]. PMID- 2554016 TI - [Advances in the biochemical and immunohistochemical study of kidney tubule transport]. PMID- 2554017 TI - [Progress in methods of studying renal tubular transport. Renal epithelial cell culture]. PMID- 2554018 TI - [Transport of chloride, bicarbonate, and proton in the proximal tubules]. PMID- 2554019 TI - [Calcium and phosphate transport by the proximal tubules]. PMID- 2554020 TI - [Regulation of Na, K transport in collecting duct]. PMID- 2554021 TI - [Cl-, HCO3-, and H+ transport in collecting duct]. PMID- 2554022 TI - [Hormone receptors and signal transduction mechanisms in renal tubules]. PMID- 2554023 TI - [Kidney tubular transport disorders. Renal tubular acidosis]. PMID- 2554024 TI - [Kidney tubular transport disorders. Vitamin D resistant rickets]. PMID- 2554026 TI - Histological effects of endocrine therapy for prostatic cancer in relation to clinical course. AB - In order to evaluate histological changes in cases of prostatic cancer following endocrine therapy, 25 extensive specimens were removed from prostates during periods of local control, and were examined with respect to prognosis. Shortly after the commencement of the endocrine therapy, there were noticeable degenerative changes in the cancer cells as well as structural changes such as desquamation of cells and loss of cancer nests. Later than two months from the start of therapy, stromal changes such as fibrosis and scar formation appeared. Coagulation necrosis of tumor tissue, inflammatory cell infiltration and granulomatous reactions were not as prominent. Within the first two months of treatment, the endocrine therapy uniformly affected the cancer tissues to some extent. After then, some showed relapsing viable cells in a part of the tumor, being judged to be no response. The others continued to respond to the therapy. The response was estimated as marked when such therapeutic changes appeared diffusely and profoundly over the tissues removed. Patients with a marked response had a good prognosis, indicating histological evaluation after endocrine therapy to provide a prognostic factor. PMID- 2554025 TI - Antitumor activity of 7-N-(2-((2-(gamma-L-glutamylamino) ethyl) dithio) ethyl) mitomycin C (KW2149) against human tumor xenografts serially transplanted into nude mice. AB - The antitumor activity and toxicity of 7-N-(2-((2-(gamma-L-glutamylamino) ethyl) dithio) ethyl) mitomycin C (KW2149) were evaluated using a human tumor xenograft- nude mouse system, and compared with those of the maternal compound, mitomycin C. The maximum tolerated dose of KW2149 was estimated to be 15 mg/kg by bolus intraperitoneal or intravenous injection, at which a remarkable reduction of spleen weight was observed, suggesting bone marrow suppression by this agent. A bolus injection of KW2149 seemed to be more effective than a divided injection schedule, when a total of 15 mg KW2149/kg was administered to mice bearing breast (MX-1) and colon (Co-4) carcinomas. The antitumor activity of KW2149 was dose dependent, and the difference in antitumor effect according to route of administration was minimal. The antitumor spectrum of KW2149 was essentially identical to that of mitomycin C administered intraperiotoneally as a bolus at a dose of 6 mg/kg. PMID- 2554027 TI - Poor prognoses following left hepatic trisegmentectomies for cancer. AB - Five patients with hepatocellular carcinoma, and one with a gallbladder cancer with liver metastases, underwent left hepatic trisegmentectomy with, in four cases, resection of the left caudate lobe. A bleeding peptic ulcer and an anaphylactic shock due to a drug allergy caused two hospital deaths. A third major complication was a prolonged bile leakage which healed spontaneously. One patient who died in hospital had not undergone a radical resection, and all four patients surviving the procedure eventually died with recurrent local (and sometimes also distant) tumors between 3.5 and 11 months after resection. The results appear to give relatively few indications for left trisegmentectomy in the treatment of these tumors. PMID- 2554028 TI - A case of the watery diarrhea-hypokalemia-achlorhydria syndrome: successful preoperative treatment of watery diarrhea with a somatostatin analogue. AB - A 35-year-old man presenting with severe watery diarrhea was diagnosed as having the watery diarrhea, hypokalemia and achlorhydria (WDHA) syndrome with the elevation of plasma vasoactive intestinal peptide (VIP) level. Imaging diagnostic techniques revealed a hypervascular tumor at the tail of the pancreas as well as a solitary liver metastasis. During the patient's stay in hospital, he developed acute renal failure probably due to persistent dehydration and severe hypokalemia. Although these complications improved with artificial dialyses, severe watery diarrhea continued, which made it difficult to achieve surgical resection of the tumor. A new long-acting and potent somatostatin analogue, SMS 201-995 (Sandoz Ltd, Basel, Switzerland), was tested and was shown to be effective; after a few hours of subcutaneous injection of this agent, the watery diarrhea disappeared, which in turn improved the patient's hypokalemia, hypercalcemia and metabolic acidosis. Three weeks later, distal pancreatectomy with splenectomy and hepatic lobectomy were successfully performed, and the patient resumed his normal life. The somatostatin analogue has been reported to be useful in the long-term treatment of patients with inoperable WDHA syndrome. The present case demonstrated that short-term administration of this agent is also useful for improving the condition of WDHA patients at the preoperative stage. PMID- 2554029 TI - [Angiographic features of malignant fibrous histiocytomas]. AB - The angiographic findings in eight cases with malignant fibrous histiocytomas are described. These lesions were well demarcated and demonstrated fine tumor vessels, homogenous stains with avascular areas presenting central necrosis. But these angiographic findings could not differentiate MFH from synovial sarcoma and fibrosarcoma. PMID- 2554030 TI - [Postoperative compensatory hypertrophy of the liver imaged by liver scintigraphy and CT]. PMID- 2554031 TI - Lymphokine mediated microbicidal activity of peritoneal macrophages from Leishmania donovani infected and drug treated BALB/c mice. AB - The resident peritoneal macrophages from untreated mice develop potent microbicidal activity against amastigotes of Leishmania major and Leishmania donovani after in vitro exposure to lymphokines (LK) from mitogen stimulated spleen cells. However, to the best of our knowledge, the response of L. donovani infected peritoneal macrophages from already infected/treated animals to LK has not been investigated. Therefore in the present study, the effect of LK on infected macrophages from BALB/c mice following specific infection and subsequent treatment with sodium stibogluconate has been investigated. As the infection progressed, a decrease in percent microbicidal activity was noticed. An attempt was also made to treat the animals on different post infection days and reinfect them in vitro. Infection could not be produced in vitro in late treatment groups when the treatment was given on 14 days and 21 days post infection. Whereas, macrophages obtained from animals treated on 7 days post infection (early treatment) could be infected in vitro. However, only 50% of the cells got infection. This infection was eliminated when the cells were exposed to LK for 72 hours. PMID- 2554032 TI - Elevation of laminin and beta-subunit of prolyl 4-hydroxylase in the sera of human subjects with Schistosomiasis mansoni. AB - Serum levels of several markers for liver fibrosis were measured utilizing three groups of human subjects related with schistosomiasis mansoni in northeast Brazil; (1) 20 Schistosoma mansoni egg-positives, who have never been administered with anti-schistosomal drugs, (2) 29 egg-negative inhabitants in the endemic area of schistosomiasis, and (3) 23 egg-negative Japanese immigrants in the non-endemic area. None of these sera were positive for antibody to the surface antigen of human hepatitis B (HBs) and circulating HBs antigen. There was no significant difference in the serum levels of N-terminal peptide of procollagen type-III between the egg-positive subjects and either of the egg negative Brazilian or Japanese immigrants, whereas the mean value of serum laminin significantly increased in the egg-positive subjects. A significantly higher concentration of serum immunoreactive beta-subunit of prolyl 4-hydroxylase (IR beta PH) was also observed in the egg-positive subjects only in comparison with that of the egg-negative Brazilian. Serum laminin and IR beta PH concentrations of the egg-positive subjects did not correlate with the absorbance of enzyme-linked immunosorbent assay (ELISA) which utilized crude antigens isolated from schistosome adults or eggs. No significant difference in these two parameters was observed between two subgroups of the egg-negative Brazilian or Japanese immigrants divided according to the serological data by ELISA. These findings suggest that serum laminin and IR beta PH levels are worth further evaluation for their usefulness as the marker for liver fibrosis in schistosomiasis. PMID- 2554033 TI - The effect of prostacyclin treatment on the changes of cyclic nucleotide levels of rat antral and fundic gastric mucosa in different experimental ulcer models. PMID- 2554034 TI - [An immunological and clinical evaluation of combined TAE-LAK adoptive immunotherapy]. AB - To clarify the immunological effects of transcatheter arterial embolization (TAE) therapy for hepatocellular carcinoma (HCC), various immunological parameters were measured before and after TAE respectively. In most effective group of which AFP levels at first week after TAE therapy had decreased more than 50% compared with those before TAE, the percentage of OKT-4 and IL-2R positive cells in the peripheral blood lymphocytes (PBL) had significantly increased in number. In addition, IL-1, TNF, IL-2 and LAK activity were also enhanced by it. These immunological enhancement after TAE therapy was suggested to be a favourite condition for transferring LAK cells to the patients with HCC. Therefore the combination therapy with TAE and LAK-adoptive immunotherapy was conducted in 12 patients with HCC. Partial response to it was obtained in one case and minor response in three. However, no effectiveness was also found in eight (Progressive Disease: 1 case, No Change: 7 cases). Immunological response after combined TAE LAK adoptive immunotherapy revealed that NK activity and LAK activity were markedly enhanced. Furthermore, the percentage of OKT-11+, IL-2R+ and Leu-7- 11c+ cells in the PBL had increased in number with statistically significant differences and OKT-8+ cells had increased in relative number. In conclusion, this study suggested that this combination therapy might be a well designed immunological and clinical therapy for HCC because it was done under the condition of well enhanced immunological parameters against tumors. PMID- 2554035 TI - The decomposition and aggregation of rat lens protein induced by selenite in vitro and in vivo. AB - A single subcutaneous injection of sodium selenite at the dose of 20 mumol/kg body weight induced bilateral nuclear cataracts in suckling rats. This selenite induced cataract incidence can be increased by pretreating animals with a glutathione synthesis inhibitor. Sodium dodecyl sulfate polyacrylamide electrophoresis of urea-soluble proteins from selenite-induced cataractous lenses showed the appearance of high molecular weight aggregates and decomposed products of lens proteins. These products were found in association with the emergence of a 45 K band. Incubation of water-soluble lens proteins with selenite in vitro produced changes similar to those demonstrated in selenite-induced cataractous lenses. Furthermore, selenite induced the gradual development of opalescence and the oxidation of sulfhydryl in the lens protein solution. Therefore, we presume that the oxidation of lens protein sulfhydryl by selenite is associated with both aggregate formation and the decomposition of lens proteins, and that these changes may provide a partial explanation for the mechanism of selenite cataract. PMID- 2554036 TI - [The quantitative determination of cyanide by FTD-GC]. AB - In recent buildings, new materials containing chemical goods, chemical fibers and the like are in common use. Some of them evolve hydrogen cyanide (HCN) when burning. When a person meets a fire, therefore, it is necessary to measure his blood concentration of HCN as well as that of CO-Hb%. The blood concentration of HCN can be measured by colorimetry such as pyridine-pyrazolone method, gas chromatography, electrode method, or the like. However, these methods require much time in pretreatment and the preparation of the reagent. We have investigated to find a new measurement method free from the above demerits, so that we have found the method by FTD-GC (Flame Thermionic Detecter Gas chromatograph) is convenient and can be effected in a short time in comparison with the aforementioned method. GC-7AG made by Shimadzu Co. is employed as gas chromatography and the headspace method is used in the quantitative determination. Samples are employed as solutions of potassium cyanide (KCN) either in distilled water or in fresh blood. The concentration of KCN is gradient in the range of 0.5-6 micrograms/ml. The quantitative conditions is investigated by a solution in distilled water as standard. Using this method, the concentration of HCN in the blood sample is determined slightly lower than that in the distilled water; however, this method is advantageous in convenience and the saving of time. Therefore, the method is useful in practice for the determination of the concentration of HCN.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554037 TI - [Clinical trial of 111In-antimyosin antibody imaging: (3) Comparison with 99mTc pyrophosphate imaging]. AB - Clinical value of 111In-antimyosin monoclonal antibody F ab (AM) was compared with 99mTc-pyrophosphate (PYP) in 13 patients with myocardial infarction and 3 patients with myocarditis. Following PYP injection, PYP imaging was performed 3 hours later. Immediately after PYP imaging, AM was administrated and AM images were obtained 48 hours later. Abnormal accumulation in the infarcted myocardium was observed in 11 patients (85%) on AM images but only in 3 patients (23%) on PYP images. All patients within 8 days after the onset of infarction showed abnormal uptake on both images. Of 5 patients with 1 to 2 weeks after the onset of infarction, abnormal uptake was observed in all of them on AM images but only in one of them on PYP imaging. Furthermore, of 6 patients with more than 2 weeks after the onset, AM imaging showed abnormal uptake in 4 (67%) but PYP imaging did not show abnormal uptake in any of them. Similarly. Of 3 patients with myocarditis, diffuse uptake in the myocardium ws observed in 2 of them on AM images but none of them showed abnormal uptake on PYP images. We conclude that AM imaging is a useful means for identifying not only acute stages but also subacute stages of myocardial necrosis where PYP imaging did not show any abnormality. PMID- 2554038 TI - [A case of mixed mullerian tumor, usefulness of 67Ga-citrate scintigraphy]. AB - Mixed Mullerian Tumor (M.M.T.) is a rare malignant neoplasm usually arising from uterine body in postmenoposal woman, and imaging diagnosis for the tumor has not been established. A 45-year-old female with pathologically confirmed M.M.T. was evaluated by 67Ga-citrate scintigraphy and CT on the basis of imaging diagnosis. 67Ga-citrate scintigraphy detected correctly not only primary lesions but also metastatic and recurrent-foci. The findings of 67Ga-citrate scintigraphy were confirmed to be correspond to autopsy findings. Generally, detectability of 67Ga citrate scintigraphy for genito-urinary tract neoplasm was not appreciated, but in our case 67Ga-citrate scintigraphy was useful for detection of the primary lesion, metastatic and recurrent foci. The cellular components and grade of cell differentiation of this tumor seemed to be responsible for good detection of the tumor. Thus, 67Ga-citrate scintigraphy was considered to be useful diagnostic imaging modality of M.M.T. PMID- 2554039 TI - Altered responses of plasma 18-hydroxycorticosterone and aldosterone to angiotensin II and adrenocorticotropin in patients with a 18 hydroxycorticosterone-producing tumor. AB - Plasma 18-hydroxycorticosterone (18-OHB) and aldosterone responses to angiotensin II (AII) and ACTH were examined in 2 patients with a 18-OHB-producing tumor (18 OHBPT) versus those in 8 patients with a aldosterone-producing adenoma (APA), 7 patients with low renin essential hypertension (LREH) and 10 normal subjects. Plasma 18-OHB and aldosterone levels and the 18-OHB: aldosterone ratio were high in patients with an APA and normal in patients with LREH. In patients with a 18 OHBPT, plasma 18-OHB and aldosterone levels were high and normal, respectively, resulting in a 2-fold greater 18-OHB: aldosterone ratio than that in patients with an APA. Patients with an APA had a blunted response of plasma 18-OHB and aldosterone to AII and a supranormal response of these corticoids to ACTH. Patients with LREH had a supranormal response of plasma 18-OHB and aldosterone to AII and a normal response of these corticoids to ACTH. In patients with a 18 OHBPT the responses of both plasma 18-OHB and aldosterone to AII and ACTH closely resembled those in patients with an APA but not in patients with LREH. These data suggest that 18-OHBPT may be a variant of aldosteronomas, producing a large amount of 18-OHB and a small amount of aldosterone. PMID- 2554040 TI - Biochemical changes in airways after antigen inhalation. AB - These results suggest that activation of phospholipase A2 after antigen exposure triggers an increase in leukotriene synthesis resulting in inflammatory cell influx into the airway and contraction of bronchial smooth muscle. That is, it was demonstrated that inhaled antigen concerns with IgE-mast cell system, but, on the other hand it might act as non specific stimuli. PMID- 2554041 TI - Effects of chlorpromazine in the nucleus reticularis lateralis on the cat cerebellar potentials. AB - In the experiments reported here, we investigated chlorpromazine (CPZ)-induced (CPZ)-induced enhancement of the cat cerebellar potentials evoked by peripheral nerve stimulation, with regard to the subtypes of adrenoceptor in the nucleus reticularis lateralis (LRN). Electrical stimulus of the locus coeruleus (LC) at high frequencies enhanced cerebellar potentials evoked by peripheral nerve stimulation. Although similar stimulus increased them after pretreatment with an alpha 2-antagonist, yohimbine, these enhancements were not recognized by pretreatment with an alpha 1-antagonist, prazosin. Microinjection of norepinephrine (NE: 10 micrograms) into LRN decreased cerebellar potentials, and conversely, 30 micrograms of NE significantly increased them. Although microinjection of an alpha 2-adrenoceptor agonist, clonidine, into the LRN depressed cerebellar potentials, clonidine-induced decrease was obviously antagonized by pretreatment with CPZ. Furthermore, an alpha 1-adrenoceptor agonist, phenylephrine, into the LRN increased cerebellar potentials. Pretreatment with CPZ hardly changed phenylephrine-induced enhancement. We though that CPZ blocked alpha 2-autoreceptors in adrenergic terminals from the LC rather than alpha 1-adrenoceptors within the LRN. As a result, NE released from the LC terminals may act on alpha 1-adrenoceptors in the LRN and may attenuate activities of the LRN. Therefore, it was clear that previous CPZ-induced enhancement may be due to depression of the descending inhibitory adrenergic system via CPZ-induced blockade of alpha 2-autoreceptors. PMID- 2554042 TI - A tumor-associated antigen in the scirrhous gastric carcinoma cell line MK-01 defined by monoclonal antibody S202. AB - A monoclonal antibody (MAb) S202, with an IgG1 isotype, that reacted strongly with the scirrhous gastric carcinoma cell line MK-01 was established. MAb S202 reacted with the colonic cancer cell line, SW1116, and the pancreatic cancer cell line, PK-1, when tested by indirect immunofluorescence. The S202 reactive antigen was expressed in the majority of acetone-fixed fresh frozen cancer tissues. Eighty to 100 per cent of the paraffin-embedded sections of stomach, colon and pancreatic adenocarcinoma were positive for the S202 antigen, with diffuse cytoplasmic staining, whereas esophageal and breast cancers demonstrated markedly less immunostaining. Supplemented serum-free medium collected from 7 day old tumor cell cultures were assayed for the presence of antibody-defined antigens. Antigens detected by MAb S202 were released by the cell lines SW1116 and PK-1. The binding of MAb S202 to the colonic adenocarcinoma sections was reduced after treatment with sodium periodate which suggests that respective antigenic determinants are of carbohydrate nature without sialic acid residues. PMID- 2554044 TI - [Silicosis and lung cancer]. AB - A clinicopathological analysis was carried out on 50 cases of lung cancer with silicosis diagnosed from April 1975 to March 1988. All patients were males and the age distribution ranged from 47 to 85 years with a mean of 63.5 at diagnosis. They had been exposed to silica in tunneling for 3 to 42 years, with an average of 15.1. Forty eight cases smoked. Histologically, squamous cell carcinoma was the most common with 29 cases, followed by 10 small cell carcinomas, 6 adenocarcinomas, 4 large cell carcinomas and one adenosquamous carcinoma. Thirty seven tumors were located in peripheral regions, mostly upper lobe or S6, while 13 tumors were in large bronchi. As the most common histological types of lung cancer with silicosis were squamous cell carcinoma and small cell carcinoma, some carcinogens might be involved in tumorigenesis. Silica alone is not considered to be a carcinogen, however, silica containing adsorbed polycyclic aromatic hydrocarbons from cigarette smoking or from industrial pyrolysis can act as a carcinogen or promoter. PMID- 2554043 TI - A case of hepatoblastoma occurring in an adult. AB - We present herein a rare case of hepatoblastoma occurring in an adult male. The patient was 22 years old and his laboratory investigations on admission showed a marked elevation of alpha-fetoprotein in the serum. CT scan and other examinations revealed a primary tumor, 6.5 cm in size, in the left hepatic lobe with metastasis in the head of the pancreas. Thus left hepatic lobectomy and pancreaticoduodenectomy were performed, but metastasis to the right hepatic lobe, left lung and abdominal skin were found 2 months later. Despite repeated courses of chemotherapy with adriamycin and cisplatin, the patient died 9 months after his operation. Pathological findings revealed poorly differentiated type hepatoblastoma. A review of the literature revealed only twelve other such cases. PMID- 2554045 TI - [Clinical evaluation of surgical treatment of lung cancer involving the chest wall]. AB - Twenty-four patients with primary lung cancer involving chest wall underwent pulmonary and chest wall resections. All patients were males ranging from 38 to 74 years of age. Pain was the most frequent symptom. Twenty-three lobectomies and one pneumonectomy were performed. Resections of parietal pleura were performed in 4 patients and en block resections of chest wall were performed in 20 patients. After operation, postsurgical stage was pT3N0M0 in 16 patients, pT3N1M0 in 4 patients and pT3N2M0 in four. Operative mortality was 4.2%. Actuarial survival of 23 patients surviving operation was 45.7% at 1 year, 28.7% at 2 year and 17.2% at 3 and 5 year. Three patients who had no lymphnode metastases and no rib invasion survived more than 5 years. We concluded that long term survival can be expected in pN0 patients without invasion to the ribs. PMID- 2554046 TI - [In vitro susceptibility testing of nontuberculous mycobacteria to streptomycin, kanamycin and enviomycin]. AB - In vitro susceptibility testing to streptomycin, kanamycin and enviomycin was carried out in nontuberculous mycobacteria and the results were compared with those of M. tuberculosis strains. Among nontuberculous mycobacteria tested, only M. xenopi strains exhibited similar minimal inhibitory concentration (MIC) values to streptomycin with M. tuberculosis strains. On the other hand, M. xenopi, M. malmoense and M. marinum strains showed the same MIC values to kanamycin with the MIC values for M. tuberculosis strains. The MICs of enviomycin for M. kansasii, M. malmoense, M. szulgai, M. xenopi and M. marinum strains were similar to those for M. tuberculosis strains. Out of 64 strains of M. avium complex, the MIC value similar to that for M. tuberculosis strains was shown in 16% of strains to streptomycin, in 42% of strains to kanamycin, and in 50% of strains to enviomycin. PMID- 2554047 TI - [Membrane-protective effect of finoptin in exertion-induced stenocardia]. AB - Ninety five males of employable age who suffered from effort-associated angina were examined. As the severity of angina increased, the structural disorganization of the lipid bilayer progressed in the erythrocyte membranes, resulting in abnormal functional status of Na+,K+- and Ca2+-ATPases and in accumulation of Ca2+ and Na in the intracellular space, by lowering potassium levels. Finoptin given to patients with angina not only diminished intracellular calcium concentrations, but produced membrane-protective effects by lowering malonic dialdehyde levels in the lipid bilayer of the cellular membranes, by accumulating phospholipids and cholesterol esters, and returning the values of Na+,K+- and Ca2+-ATPase activities to controls. A parallelism was found between the magnitude of the membrane-protective action of finoptin in patients with angina and its antianginal effect. PMID- 2554048 TI - LTB4 in nephrotoxic serum nephritis in rats. AB - We studied leukotriene B4 (LTB4) synthesis in isolated glomeruli of rats with nephrotoxic serum nephritis. This nephritis was induced in male Sprague Dawley rats by injecting one proteinuric dose of nephrotoxic serum (rabbit anti-rat-GBM serum) after prior immunization of the rats with rabbit IgG. Histological and analytical examinations were performed in kidneys perfused until free of blood 6, 12, 24, 48 and 72 hours after induction of the disease. To investigate LTB4 production, glomeruli were isolated and incubated for one hour in the presence of Ca++-ionophore A23187. The supernatants were analyzed for LTB4. The peak comigrating on reverse-phase high performance liquid chromatography (RP-HPLC) with reference LTB4 was isolated. The ethyl ester trimethylsilyl ether derivative of this peak was analyzed by gaschromatography-mass spectrometry (GC/MS). Identical spectra of the glomerular samples and of reference LTB4 in the positive and in the negative ion chemical ionization mode provided unequivocal evidence that the substance released from the nephritic glomeruli was indeed LTB4. Six hours after injection of nephrotoxic serum, glomerular LTB4 release was highest with 5.52 +/- 0.50, then declining to 2.20 +/- 0.10 ng/mg glomerular protein at 12 hours. At 24, 48 and 72 hours no statistically significant difference from control animals was found. No metabolism of LTB4 to 20-hydroxy- or 20-carboxy LTB4 was detected during the incubation period. Albuminuria developed during the first 24 hours after nephrotoxic serum challenge and rose steadily throughout the observation period up to 277 +/- 25 mg/24 hr after 72 hours.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554049 TI - Blunted excretory response to atrial natriuretic peptide in experimental nephrosis. AB - Adriamycin (ADR) nephrosis and a model of unilateral ADR-induced proteinuria were produced in Sprague-Dawley (S.D.) rats to investigate the mechanism of sodium retention by the nephrotic kidney. Plasma volume, as measured by the dilution principle using radioiodinated serum albumin, was significantly higher in nephrotic animals than in control ones (NS: 69.61 +/- 15.02: control: 47.05 +/- 5.32 ml/kg: P less than 0.01). Similarly plasma levels of immunoreactive ANP (iANP) were significantly higher in nephrotic animals compared to controls (NS 104.22 +/- 36.41: control 59.94 +/- 20.88 pg/ml; P less than 0.05). Using the unilateral model we found a markedly reduced diuretic and natriuretic response to the infusion of synthetic rat atrial natriuretic peptide (ANP 1-28) in proteinuric kidney but not in contralateral kidney, despite a comparable increase in glomerular filtration rate. To explain the blunted diuresis and natriuresis in the presence of normal glomerular response to ANP, we investigated the possibility of an abnormality at post-glomerular level by studying ANP receptor density and affinity of the inner stripe of outer medulla and the inner medulla in ADR-and vehicle-treated rats. The inner stripe of outer medulla and the inner medulla receptor density and affinity were not significantly different in ADR rats as compared to animals given the vehicle alone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554050 TI - Regulation of the renal Na+-H+ exchanger by protein phosphorylation. AB - Starting from observations in intact cells and extending to studies in native membranes and solubilized membrane proteins, a significant body of evidence has been accumulated to indicate that some of the short-term regulatory influences on the Na+-H+ exchanger in the apical membrane of the proximal convoluted tubule act via protein phosphorylation mediated by specific protein kinases. Protein phosphorylation mediated by PKA inhibits the Na+-H+ exchanger while that mediated by PKC stimulates activity. The effect of PKA and PKC on the Na+-H+ exchanger in native membranes and in solubilized brush border membrane proteins appears to be consistent with most of the published observations in intact cells. Further studies using solubilized, renal brush border membrane proteins indicated that protein phosphorylation mediated by CaM-kinase II inhibited the activity of the Na+-H+ exchanger. The physiologic significance of this observation in intact cells remains to be determined. It is hoped that the types of experimental approaches outlined in this review will yield additional insights into the structure of the Na+-H+ exchanger and to a clearer understanding of its physiologic regulation. PMID- 2554051 TI - Increased Na+/H+ antiport activity in the renal brush border membrane of SHR. AB - Defect in renal salt excretion may play an important role in the pathogenesis of hypertension. We examined sodium (Na+) uptake by brush border membrane (BBM) vesicles of young (6 week old) spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY) of the same age. SHR had lower urinary Na+ excretion (223.1 +/- 9.3 vs. 266.3 +/- 3.7 microEq/day/100 g, N = 8, P less than 0.01) and higher systolic blood pressure (98.9 +/- 1.2 vs. 82.9 +/- 1.8 mm Hg, N = 8, P less than 0.01) than WKY. BBM vesicle Na+ uptake, measured by rapid filtration technique, was higher in SHR when compared to WKY (1.44 +/- 0.03 vs. 1.01 +/- 0.06 nmol/mg/5 sec, N = 4, P less than 0.01). This increase in Na+ influx was apparent only in the present of an outward-directed proton (H+) gradient and was abolished by 1 mM amiloride. BBM permeability to H+ as assessed by acridine orange quenching was not different between SHR and WKY. Kinetic analyses of the amiloride-sensitive BBM Na+ uptake revealed a higher Vmax (2.13 +/- 0.27 vs. 0.70 +/- 0.30 nmol/mg/5 sec, N = 4, P less than 0.01) and a higher km for Na+ (3.55 +/- 0.32 vs. 1.23 +/- 0.14 mM, N = 4, P less than 0.05) in SHR. These findings thus demonstrate an intrinsic derangement in BBM Na+ transport in young SHR which is characterized by increased Na+/H+ antiport activity. This alteration in antiport activity is not attributable to changes in membrane permeability to H+, and is characterized by higher Vmax and km.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554052 TI - Urokinase synthesis and binding by glomerular epithelial cells in culture. AB - Fibrin deposits are frequently observed in the course of proliferative extracapillary glomerulonephritis and could be related to a defective local fibrinolysis. We studied human glomerular epithelial cells in culture which were found to release mainly a urokinase-type plasminogen activator (u-PA) identified on zymography by its molecular weight (53 kD), its plasminogen activator activity, and its neutralization by specific polyclonal anti-u-PA IgG. Trace amounts of tissue-type plasminogen activator (t-PA) complexed to a plasminogen activator inhibitor type 1 (PAI-1) were identified with specific antibodies. Specific binding sites were found at the surface of glomerular epithelial cells (kD: 2.10(-9) M), partially occupied by secreted u-PA. The spontaneous u-PA activity of the culture medium conditioned by glomerular epithelial cells was very low, suggesting that u-PA was released in its inactive single chain proenzyme form (SC-u-PA). After activation of SC-u-PA by plasmin, u-PA activity of the culture medium was found to increase in a time- and dose-dependent manner when cells were incubated with phorbol myristic acetate (PMA). This effect was inhibited by H7, a protein kinase C inhibitor. Stimulation of u-PA synthesis by PMA was also observed in two different epithelial tubular cell lines. LLC-PK1 and MDCK cells. However, 8 bromo cyclic AMP which increased u-PA release by LLC-PK1 cells was found to inhibit u-PA release by PMA-stimulated glomerular epithelial cells and MDCK cells. By Northern blot analysis we found that PMA induced an increase of u-PA mRNA level in glomerular epithelial cells and that cyclic AMP had an opposite effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554053 TI - Effects of antihypertensive drugs on glomerular morphology. AB - We quantitated the glomerular size and the degree of sclerosis simultaneously in individual glomeruli with the use of three-dimensional histological analysis on serial sections obtained from remnant kidneys with highly heterogeneous glomerular lesions after subtotal nephrectomy (sNPX). Four to six weeks after sNPX (Group I, N = 7), 90% of glomeruli had mild sclerosis (sclerosis index, SI; less than 1.5 on a 0 to 4 scale) with a strong positive correlation between the maximum planar area of glomerulus (PAmax) versus SI. Twelve weeks after sNPX (Group II, N = 6) more than 50% of glomeruli had advanced sclerosis (average SI:1.88), and a significant positive correlation was again found between PAmax and SI in glomeruli with mild to modest sclerosis (SI less than 1.5), whereas these two variables were correlated inversely in glomeruli with advanced sclerosis. Administration of enalapril (50 mg/liter drinking water) or hydralazine (200 mg/liter) + hydrochlorothiazide (50 mg/liter) for 12 weeks (Group III, N = 12) markedly attenuated the sclerosis to comparable degrees (average SI: 0.15 vs. 0.22). The former antihypertensive therapy decreased glomerular capillary hydraulic pressure (PGC) to normal range, whereas the latter triple drug therapy was largely without effect on PGC. Of note, the positive correlation between SI and PAmax remained unaffected by these anti-hypertensive drugs. SI of the glomeruli from both treated groups was expressed as a first order function of PAmax. The correlation coefficient is identical to that found in non-treated Group II remnant glomeruli, so that the degree of sclerosis is mathematically uniquely correlated with the glomerular size, regardless of drug treatment. Thus, within a given remnant kidney, the magnitude of glomerular hypertrophy has a direct correlation with the degree of sclerosis, while the altered glomerular hemodynamic pattern has little modulatory role in determining the magnitude of this hypertrophy. Enalapril and triple drug therapy, at equi depressor doses in regard to systemic blood pressure, had identical potency in sparing glomerular structure. The primary determinant for this antisclerotic potency appears to be related to the drugs' potency to inhibit glomerular growth rather than an effect on the abnormal hemodynamics which develop in the glomerulus. PMID- 2554054 TI - [Debatable issues in the diagnosis of angiotrophic neuropathies of the limbs]. AB - On the basis of their rich personal material and the data in the literature, the authors point out the tendency to relate many types of angiotrophic neuroses to Raynaud's phenomenon disregarding their etiological and pathogenetic factors. Diagnostic errors occur particularly frequently in chronic chilblain of the limbs and its sequelae which are manifested by a characteristic complex of symptoms, successively changing vasomotor reactions in the distal parts of the limbs, the development of vegetative polyneuritis, and in severe cases also by affection of the motor apparatus of the hands. The disease is encountered mostly in workers working in the open air in subnormal temperature of the surroundings. The object of the communication is to focus attention on the groundlessness of the diagnosis of Raynaud's disease which is not a nosological form. PMID- 2554055 TI - Physiology and molecular biology of the renal Na/H antiporter. AB - The renal Na/H antiporter is involved in cell pH regulation and, predominantly in the proximal tubule, also mediates transcellular NaCl and NaHCO3 transport. The transporter is regulated by systemic acid-base factors and hormones such as angiotensin II, PTH, glucocorticoids and alpha2-adrenergics. The human gene of the Na/H antiporter has been cloned and sequenced recently. PMID- 2554057 TI - Influence of viral infections on body weight, survival, and tumor prevalence in Fischer 344/NCr rats on two-year studies. AB - Sendai virus (SV), pneumonia virus of mice (PVM), and rat coronavirus/sialodacryoadenitis virus (RCV/SDAV) were common viral infections of rats in the National Cancer Institute-National Toxicology Program (NCI-NTP) studies from 1977 to 1983. Influence of these viral infections on body weight, survival, and prevalences of spontaneous tumors in the F344/NCr rats of 28 diet control groups at five different laboratories were evaluated. Tumor prevalences evaluated in this investigation included the following: leukemia and tumors of the anterior pituitary, lungs, salivary glands and Harderian glands in both sexes; adrenal pheochromocytomas in male rats; and mammary tumors in female rats. SV and PVM but not RCV/SDAV infections were associated with significant (P less than 0.05) decreases in body weights of male and female rats. Male rat groups with PVM infection had a lower prevalence of leukemia and male rat groups with RCV/SDAV infection had a higher prevalence of anterior pituitary tumors than the corresponding uninfected groups. Female rat groups with SV infection had greater survival and a higher prevalence of lung tumors than groups without SV infection. However, none of the tumor prevalence and survival differences were statistically significant when interlaboratory variability and time-related effects were taken into account. PMID- 2554058 TI - Regulation of superoxide responses of human neutrophils by adenine compounds. Independence of requirement for cytoplasmic granules. AB - Recent evidence suggests that the adenine compounds ATP, adenosine-5'-O-(3 thiotriphosphate) (ATP gamma S), and adenosine have important regulatory effects on O2- responses of human neutrophils stimulated with the chemotactic peptide N formyl-Met-Leu-Phe (fMLP). Because of evidence that receptors on neutrophils may be modified by granule fusion events, we assessed the extent to which these adenine compounds affect fMLP and CR3 (C3bi) receptors on neutrophils and whether cytoplasmic granules are required for the ability of the adenine compounds to modify O2- responses in neutrophils stimulated with fMLP. Incubation of neutrophils with ATP gamma S or adenosine led to a decrease in numbers of fMLP receptors (17 and 9.2%, respectively) but no change in receptor affinity (Kd). Paradoxically, ATP gamma S caused an increase in CR3 receptors (Mo1, CD-11b antigen), suggesting that fMLP and CR3 receptors may be under separate control. The ability of the adenine compounds to modify O2- responses in fMLP-stimulated cells was equivalent in both neutrophils and cytoplasts, suggesting that the regulatory effect of ATP, ATP gamma S, and adenosine do not require the presence of cytoplasmic granules. ATP gamma S caused enhancement of O2- responses of neutrophils to phorbol 12-myristate 13-acetate, raising the possibility that ATP gamma S may be affecting late events in the signal transduction pathway. PMID- 2554059 TI - Expression of insulin-like growth factor-II mRNA in fetal kidney and Wilms' tumor. An in situ hybridization study. AB - The pattern of insulin-like growth factor II (IGF-II) mRNA expression in developing kidney and Wilms' tumor was examined with in situ hybridization. In developing kidney, IGF-II was primarily expressed in blastemal cells and lost with their differentiation. In triphasic Wilms' tumor, a similar relationship was found. But in a monomorphous Wilms', tumor cells with epithelial differentiation expressed IGF-II mRNA. These data suggest that IGF-II may be involved in fetal nephrogenesis, that its expression is inversely coupled to normal epithelial differentiation, and that this differentiation may be aberrantly regulated in Wilms' tumor. PMID- 2554056 TI - Peptide-dependent regulation of epithelial nephron functions. AB - It has become evident that the nephron is an important target organ of many of the regulatory peptides; this brief overview will not attempt to consider the vast amount of work on peptide-dependent kidney functions; instead, it will emphasize recent work directed towards understanding intracellular signal pathways between peptide ligand-receptor interaction and expression of physiological transport responses in renal epithelial cells. The awareness that peptide hormones of differing origin, e.g., intestinal and cardiac, share at least some of the signal steps in nephron cells, has stimulated work on nephron segmental analysis of receptor binding, of second messengers, of membrane G proteins, of protein phosphorylation, and of final membrane transport responses, such as peptide-dependent ion channel regulation. Peptides involved in cell growth and differentiation, e.g., growth factors, appear to act through part of the signal pathway shared by other peptides. The peptides selected for the purpose of this review, then, are those that have been linked, by experimental evidence, to intracellular messenger systems in nephron epithelia. PMID- 2554060 TI - Tumor necrosis factor-alpha-associated uterine endothelial injury in vivo. Influence of dietary fat. AB - The influence of dietary fat on recombinant human tumor necrosis factor-alpha (rHuTNF-alpha)-associated vascular endothelial injury in mice was examined. Histopathologic evaluation showed that greater injury was consistently observed in the uterus characterized by necrosis of endothelial cells lining small vessels in the endometrium and accompanied by multifocal hemorrhage and inflammatory cell accumulation. Mice fed standard animal diet, high corn oil diet, or high menhaden oil diet showed no differences in their susceptibility to the acute injury caused by rHuTNF-alpha injected alone or in combination with recombinant murine interferon-gamma. These results indicate that uterine endothelium may be a more sensitive target for rHuTNF-alpha-mediated endothelial injury and that high menhaden oil diet does not protect against tissue injury associated with the administration of rHuTNF-alpha alone or in combination with recombinant murine interferon-gamma. PMID- 2554061 TI - Immunologic consequences of acute ethanol ingestion in rats. AB - Acute ethanol (EtOH) intoxication is commonly associated with many medical and surgical problems which primarily or secondarily involve infection. Chronic EtOH ingestion has been associated with immune dysfunction and an increased risk of infection; however, the relationship of acute EtOH exposure and immune function has not been clearly defined. To determine if there is a relationship between acute EtOH intoxication and immune function, the effects of a single EtOH ingestion on immune function were studied in a rat model. Acute intoxication was produced by gavage feeding of 3 g/kg of EtOH, and immune function was evaluated by in vivo chemotaxis to a chemotactic peptide, N-formyl-methionyl-leucyl phenylalanine and responsiveness of splenic lymphocytes to B- and T-cell mitogens. Chemotaxis was significantly suppressed at 4 hr (49.0 +/- 5.1% of control) after EtOH ingestion (P = 0.001), but returned to normal by 24 hr and remained at that level. However, acute EtOH ingestion showed no suppressive effects on lymphocyte responsiveness to either concanavalin A or LPS. These results indicate that a single ingestion of EtOH has the potential to transiently suppress chemotactic function of granulocytes but not affect lymphocyte mitogenic responsiveness in rats. This potential may contribute to increased susceptibility to infection in patients after EtOH ingestion. PMID- 2554062 TI - Regulation of ovarian function by catecholestrogens: current concepts. AB - Development of the ovarian follicle(s) destined for ovulation appears to be a process in which antral follicles undergo a recruitment, selection and subsequent dominance phase. Several intraovarian or autocrine/paracrine regulatory mechanisms have been evoked to explain these processes. One of these potential autocrine/paracrine regulators is a catecholestrogen, 2-hydroxy-estradiol (2-OH E2). Evidence implicating 2-OH-E2 as an autocrine/paracrine regulator of follicular function is reviewed. Studies have shown 2-OH-E2 to be present in nanomolar concentrations in fluid of human and equine follicles. In addition, the enzyme responsible for converting estradiol (E2) into 2-OH-E2, estrogen 2 hydroxylase (E-2-H), is abundant in granulosa and thecal cells (but not corpora lutea) of porcine follicles. Moreover, activity of E-2-H increases during follicular development in pigs. In vitro, the actions of 2-OH-E2 have been compared to those of E2, gonadotropins, catecholamines, and androgens. Studies indicated that the maximal stimulatory effects of 2-OH-E2 on progesterone production were comparable to those of E2 and gonadotropins, and greater than androgens or catecholamines. The effect of 2-OH-E2 was found to be significantly additive to each of the other classes of compounds at maximally effective concentrations, suggesting that the mechanism of action of 2-OH-E2 was different. The mode of action of the several stimulators of progesterone biosynthesis were examined additionally with antihormones for E2, catecholamines, and androgens. In each instance, the hormonal antagonists were able to inhibit the action of the predicted class of effector compounds. However, with the exception of the antiandrogen, hydroxyflutamide, no effects of anti-hormones on the action of 2-OH E2 were observed. In the aggregate, these studies suggested that the action of 2 OH-E2 is mediated by a mechanism discrete from those of the other classes of hormones examined to date, and that hydroxyflutamide exhibits both antiandrogen and anticatecholestrogen activity. 2-OH-E2 can also enhance the actions of other trophic hormones, epinephrine, LH and FSH, by enhancing hormone-stimulated cAMP production. This effect on epinephrine action appears to be due to a 2-OH-E2 stimulated increase in the density of beta-adrenergic receptors. Whether 2-OH-E2 stimulates an increase in the number of LH and FSH receptors remains to be determined. The precise locus of the stimulatory effect of 2-OH-E2 alone on steroidogenesis is unclear but preliminary data would suggest that 2-OH-E2 may be stimulating side-chain cleavage enzyme activity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2554063 TI - Anomalous oxidative cleavage of the side chain of 18-oxocortisol and its tetrahydro metabolite. AB - The course of oxidative side chain cleavage of two recently isolated 17 alpha hydroxy C-18 oxygenated naturally-occurring corticosteroids differed in that 18 hydroxycortisol yielded a 17-ketosteroid (Chu M. D., and Ulick S. (1982) J. biol. Chem. 257, 2218-2224) whereas 18-oxocortisol yielded a gamma-etiolactone (Ulick S., Chu M. D. and Land M. (1983) J. biol. Chem. 258, 5498-5502). In an analytic application of the periodic acid oxidative cleavage reaction to 18-oxocortisol, the finding of a second product of the reaction prompted a reinvestigation of its course. Both the delta 4,3-ketone secreted form of the steroid and its tetrahydro urinary metabolite were cleaved predominantly to 17-ketosteroids along with smaller amounts of gamma-etiolactones, whose proof of structure is herein reported. This anomalous course of oxidative cleavage was considered to reflect an equilibrium between C-20 ketone and cyclic-hemiketal forms in which the glycerol-type side chain structure of the later becomes the precursor of a 17 ketosteroid. Because of the similarities between in vitro oxidative side chain cleavage and in vivo corticosteroid metabolism, these findings suggest that C-18 oxygenated steroids may contribute to the 17-ketosteroid fraction of human urine. PMID- 2554064 TI - Expression of hydroxysteroid sulphotransferase is related to estrogen receptor status in human mammary cancer. AB - A positive correlation between the expression of estrogen sulphotransferase (EC 2.8: 2.4) and the estrogen receptor (ER) in human breast cancer tissues was previously demonstrated. We have now established that a similar correlation exists between the expression of hydroxysteroid sulphotransferase (EC 2.8: 2.2) and ER in such tissues. Enzyme activity was present in 93% of the ER + tumor cytosols (mean 59 +/- 44 (SD) pmol dehydroepiandrosterone sulphate formed per mg protein per 2 h (n = 42). Activity was detected in 68% of ER - tumors and this was significantly lower (mean 21 +/- 26 (SD) (n = 19), P less than 0.001) than the former group. Metabolism of estradiol-17 beta (E2) and the adrenal-derived estrogen 5-androstene-3 beta, 17 beta-diol (ADIOL), which is a substrate for hydroxysteroid sulphotransferase but not estrogen sulphotransferase, was studied in four ER + human mammary cancer cell lines (MCF-7, T47-D, MDA-MB-361 and ZR-75 1) and four ER-human mammary cell lines (BT-20, MDA-MB-231, MDA-MB-330 and HBL 100), employing steroid concentrations of 1 nM. At this concentration, formation of ester sulphates was a major route of metabolism in the ER + cell lines; E2 yielding a mean of 6.5 pmol estrogen monosulphates/mg DNA in 16 h and ADIOL yielding a mean of 9.4 pmol C19-5-ene steroid monosulphates/mg DNA in 16 h. In three of the four ER - cell lines, formation of sulphates from E2 occurred at an eight-fold lower rate (mean 0.8 pmol estrogen sulphates/mg DNA in 16 h), whereas MDA-MB-330 cells did not form estrogen sulphates. Only one of the four ER- cell lines (BT-20) sulphurylated ADIOL and this was at a 12-fold lower rate compared to the mean value for the ER + cel lines. Oxidation of E2 and ADIOL occurred in all cell lines and was generally the major route of metabolism in the ER - cells. A significant correlation between formation of estrone and dehydroepiandrosterone occurred for all cell lines (r = 0.98, P less than 0.001) indicating that the same 17 beta-hydroxysteroid dehydrogenase was probably involved. Since ADIOL is estrogenic in a number of systems at the concentration found in the blood of Western women (approximately 2 nM), the coordinated expression of hydroxysteroid sulphotransferase, estrogen sulphotransferase, and ER, supports the concept of a functional relationship between estrogen action via ER and sulphurylation reactions. PMID- 2554065 TI - Colon ornithine decarboxylase activity following standard endoscopy preparation regimens. AB - Ornithine decarboxylase (ODC) catalyzes the formation of putrescine from ornithine, which is the first step in the pathway of mammalian polyamine biosynthesis. Tissue activity levels of ODC have been suggested to be a marker of risk for colorectal cancer in hereditary polyposis and in adenoma formers. We analyzed ODC activity in rectal and sigmoid colon mucosal biopsies obtained at 10 cm and at 30 cm in 40 healthy, colon cancer risk factor-free adults following three endoscopic preparation regimens: 1) no special preparation; 2) two phosphate enemas; and 3) "Colyte" lavage preparation 12 hr previously. Levels of ODC, measured in fresh tissue, were approximately twofold higher for enema preparation vs. no preparation (for log-transformed data: sigmoid, P less than 0.0001; rectum, P = 0.0001) and for enema preparation vs. lavage (sigmoid, P = 0.0002; rectum, P = 0.008). Lavage and no preparation ODC levels were not significantly different. ODC activity levels ranged from 0.00 to 352.96 pmol/mg/hr. PMID- 2554066 TI - IL-1 dependent model of inflammation mediated by neutrophils. AB - Interleukin-1 (IL-1) mediates a number of immunologic and physiologic responses associated with inflammation. A new model to monitor the primary effects of IL-1 and potential inhibitors on inflammation has been developed, which involves unilateral injection of 300 U of highly purified recombinant human IL-1 in mouse ears. Ear thickness of IL-1 injected ears increased 7-10-fold 24 hr posttreatment, concomitant with a corresponding increase in myeloperoxidase activity, suggesting that neutrophil influx contributes to this response. Administration of nonsteroidal antiinflammatory drugs did not influence the IL-1 effect in vivo. Inhibition of phospholipase A2 activity ameliorated the IL-1 stimulated inflammation; treatment with 10 mg/kg dexamethaxone eliminated approximately 80% of increased myeloperoxidase activity compared to control values. This model provides a well-defined in vivo assay with which to quantify the systemic effects of compounds capable of altering the activity of IL-1, and the data suggest that this mechanism may explain the unique efficacy of steroids as antiinflammatories. PMID- 2554067 TI - Evaluation of heart-lung transplant recipients with prospective, serial transbronchial biopsies and pulmonary function studies. AB - The insidious development of obliterative bronchiolitis after heart-lung transplantation is thought to be due to rejection and possibly infection (cytomegalovirus). To evaluate further, we prospectively managed the last 16 consecutive heart-lung transplant recipients with serial transbronchial biopsies with lavage and pulmonary function studies as part of a surveillance protocol or as dictated by clinical presentation. A total of 123 transbronchial biopsies with lavage were performed, 77 for clinical indications (group I) and 46 for routine surveillance (group II). Results of 64 (83.1%) group I biopsies were positive for rejection or infection. Thirty-one of these biopsy specimens showed signs of rejection (29 in group I and two in group II), characterized by a perivascular mononuclear infiltrate, lymphocytic bronchiolitis, and occasionally alveolar septal mononuclear infiltrate. Forty-six serial pulmonary function tests were performed. The forced expiratory volume in 1 second (percent predicted), forced expiratory flow rate between 25% and 75% of the forced vital capacity (percent predicted), and arterial oxygen tension (millimeters of mercury) were significantly reduced from baseline values during rejection episodes: forced expiratory volume in 1 second, 75.7% +/- 20.1% versus 52.7% +/- 18.3% (p less than or equal to 0.05); forced expiratory flow rate between 25% and 75% of the forced vital capacity, 97.6% +/- 30.5% versus 49.8% +/- 22.3% (p less than or equal to 0.05); and arterial oxygen tension, 92.1 +/- 8.8 mm Hg versus 71.4 +/- 18.8 mm Hg (p less than or equal to 0.05). The fall in pulmonary function was reversible with pulse methylprednisolone. Asynchronous heart and lung rejection was documented. Of the 29 episodes of pulmonary rejection, 18 (62%) occurred asynchronously. Ten of the 16 (62%) heart-lung recipients had at least one episode of cardiac rejection. Thirteen of 16 (81%) had at least one episode of lung rejection. Serial transbronchial biopsies with lavage, as dictated by pulmonary function tests and clinical status, have guided early and more specific therapy directed against rejection and infection. With early detection, small airway dysfunction has been reversible. PMID- 2554068 TI - Effects of low-dose marine oils on intimal hyperplasia in autologous vein grafts. AB - The effects of low-dose cod-liver oil on intimal hyperplasia of vein grafts were examined in 45 adult mongrel dogs undergoing peripheral arterial reconstruction. Fifteen animals served as the control group, 15 animals were fed a fish-oil supplement containing 240 mg of eicosapentaenoic acid daily, and a further 15 animals received 480 mg of eicosapentaenoic acid daily. Segments of undistended external jugular vein were anastomosed to bilaterally divided femoral arteries. The grafts were harvested at 6 weeks and intimal thickness was measured with a computerized interactive image analyzing system. Serum cholesterol level, prothrombin time, partial thromboplastin time, bleeding time, and platelet counts were measured before the operation and at 2, 4, and 6 weeks after the operation. Plasma levels of thromboxane B2 and prostaglandin F1 alpha and serum levels of eicosapentaenoic acid were measured before and 4 weeks after the operation. Serum cholesterol level increased similarly and significantly in all animals. Serum levels of eicosapentaenoic acid rose proportionately with the oral ingestion of fish oil but did not affect coagulation parameters. Plasma thromboxane B2 and prostaglandin F1 alpha were not significantly affected by the ingestion of marine oils. Intimal thickness was 39 +/- 5 microns in the control dogs. Ingestion of 240 mg of eicosapentaenoic acid reduced intimal thickness to 24 +/- 3 microns at 6 weeks (p less than 0.01). Increasing the dose by a factor of 2 did not decrease intimal thickness further, the intima being 23 +/- 2 microns (p less than 0.005). Our data indicate that small doses of fish oil will reduce intimal proliferation in autologous vein grafts and that marine oils may exert their beneficial effects on intimal hyperplasia by a mechanism other than their known effects on prostanoid metabolism. PMID- 2554069 TI - Heart-lung transplantation: the initial Arizona experience. AB - Since November 1985, cardiopulmonary transplantation has been performed at the University of Arizona heart transplant program. Seven patients, five women and two men, have undergone heart-lung transplantation. Five patients had primary pulmonary hypertension, and two patients had Eisenmenger's complex. The mean age was 31 years (range, 17 to 43 years). Average follow-up was 15 months (range, 3 to 34 months), with a total of 115 patient-months. There have been no operative or late deaths. Immunosuppression consisted of rabbit antithymocyte globulin, cyclosporine (Cyclosporin A), azathioprine, methylprednisolone, and prednisone. Our first five patients were aggressively diagnosed and treated for rejection by endomyocardial biopsy, with each patient having one or several treatments for acute rejection. These five patients had one or several episodes of severe infection, particularly cytomegalovirus. In our last two patients we omitted routine heart biopsies. Only those rejection episodes diagnosed by chest x-ray films are considered significant. Our last two patients have not been treated for acute rejection and have had no infections. Presently our immunologic surveillance consists only of careful clinical examination and frequent chest x ray films. Any changes in the patient's condition are aggressively investigated, searching for infection or rejection. Two patients have been used as domino donors of their native heart. PMID- 2554070 TI - Pulmonary infiltrates after heart-lung transplantation: evaluation by serial transbronchial biopsies. AB - Since January 1988, prospective serial transbronchial lung biopsies have been performed as a diagnostic procedure to facilitate the care of recipients of heart lung transplants. Eighty-five cardiac and 70 transbronchial lung biopsies have been prospectively performed in 10 patients beginning within the first week of transplantation. Forty-eight percent (34/70) of the transbronchial lung biopsies and 16.5% (14/85) of the heart biopsies were positive for either rejection or infection. Pulmonary rejection was evident by a perivascular lymphocytic infiltrate that cleared with pulse steroid therapy. Pulmonary and cardiac rejection were present synchronously on six occasions and asynchronously on 16 occasions (nine pulmonary and seven cardiac). Four patients had early cytomegalovirus pneumonitis on biopsy specimen and were successfully treated with ganciclovir. Of the 40 clinically indicated biopsies, 29 (72.5%) were positive for rejection or infection and guided subsequent therapy. In summary, transbronchial lung biopsies have provided prompt (within 24 hours) serial diagnostic information that has guided successful treatment of infection (cytomegalovirus, aspergillosis, and Pneumocystis) and rejection. Asynchronous rejection of the heart and lungs has been conclusively demonstrated. With the early detection of rejection and infection, we are optimistic that chronic airway disease in patients with a heart-lung transplant may be reduced. PMID- 2554071 TI - Separation of isozymic forms of type II calcium/calmodulin-dependent protein kinase using cation-exchange chromatography. AB - Three distinct isozymes of a type II calcium/calmodulin-dependent protein kinase (CaM kinase II) from rat forebrain cytosol were separated using S-Sepharose cation-exchange resin. About 90% of the applied kinase activity was recovered in three protein peaks. Each isozymic form of the kinase was purified 200-300 fold by chromatography on S-Sepharose, calmodulin-affinity and gel filtration resins. All 3 forms of CaM kinase II had apparent molecular masses of 650-700 kDa, but contained variable proportions of 50 kDa and 58-60 kDa subunits. The molar ratios of the 50 kDa/58-60 kDa kinase subunits in each holoenzyme were determined by protein staining and [125I]calmodulin-binding studies and were approximately: 6/1, 3/1 and 1/1. The isozyme containing a 3/1 ratio of subunits corresponds to the predominant form of CaM kinase II in forebrain representing 70-80% of the total activity in cytosol, whereas the other forms each represent 5-10% of the total cytosolic activity. The substrate specificities and time courses of substrate phosphorylation for the isozymes were comparable. These studies provide a basis to examine regional, subcellular, and developmental differences in the isozymic forms of CaM kinase II which may subserve different neuronal functions. PMID- 2554072 TI - Expression of retinoic acid receptor-alpha mRNA in human leukemia cells with variable responsiveness to retinoic acid. AB - Retinoic acid receptor (RAR)-alpha mRNA expression was studied in a variety of myeloid leukemia cells with variable responsiveness to the induction of terminal differentiation by retinoic acid (RA). Cells from both the wild-type (wt), RA responsive HL-60 promyelocytic leukemia cell line and a selected greater than or equal to 300-fold RA-resistant subline expressed approximately equal amounts of two RAR-alpha transcripts, 4.0 and 3.1 kb in size. In wt cells, the RAR-alpha did not change during induction of granulocyte differentiation by RA or macrophagic differentiation by 12-0-tetradecanoylphorbol-13-acetate (TPA). Relative to HL-60 cells, other cultured and fresh myeloid leukemia cells expressed 2.5-fold less to equal amounts of the RAR-alpha transcripts. The relative expression in six cases of acute promyelocytic leukemia (APL; two RA-responsive; one, previously treated with 13-cis-RA in vivo, equivocally RA-responsive) and one case of acute myelogenous leukemia (AML) with promyelocytosis (RA unresponsive) was 0.91 +/- 0.14 versus 0.53 +/- 0.14 for eight cases of nonpromyelocytic AML (p congruent to 0.001). Lymphoid leukemia cells expressed 2- to 5-fold less RAR-alpha mRNA. No qualitative variations in the mRNA transcripts were observed, although the 3.1 kb transcript was relatively decreased in three cases. The RAR-alpha gene was not amplified or detectably rearranged in any DNA source, although an apparent EcoRI restriction fragment length polymorphism was observed. It is concluded (a) that the steady-state level of RAR-alpha mRNA is not tightly correlated with natural responsiveness/unresponsiveness or, in some instances, acquired resistance to RA induced differentiation and (b) that further studies are needed to determine if the mean 1.7-fold higher RAR-alpha mRNA level in APL cells could be an essential factor in the RA-responsiveness of APL cells, as primarily regulated at a different molecular level. PMID- 2554073 TI - Enzyme activities in perikaryal and synaptic mitochondrial fractions from rat hippocampus during development. AB - When pharmacological or basic neurochemical systematic characterization of mitochondrial enzymatic systems correlated to energy transduction processes is attempted, studies must be based on subcellular fractions with a high degree of purity from specific brain areas and from individual animals. Distinct populations of mitochondria heterogenous with respect to biochemical enzyme characteristics from rat brain hippocampus are described. Two mitochondrial populations were derived from synaptosomes by lysis and a third consists of free non-synaptic mitochondria. The maximum rate of some cerebral enzyme activities which are part of energy transduction (citrate synthase, malate dehydrogenase; total NADH-cytochrome c reductase, cytochrome oxidase) and amino acid metabolism (glutamate dehydrogenase) were tested on these mitochondrial populations of 8- and 16-week-old rats. A comprehensive analysis of the data suggests that extensive but highly diversified catalytic expressions of the enzymes studied occur in the hippocampus. This is true even when a short period of the rat life span is studied. Hence the varying pattern of evolution of the differing cerebral mitochondria, probably a consequence of different metabolic functions, should be taken into account in any pharmacological study on these systems. PMID- 2554074 TI - Upregulation of adrenergic beta receptor subtypes in the senescent rat heart. AB - Beta receptors in older hearts respond to procedures which cause upregulation (N. Tumer et al., J. Gerontol., 1989, in press). To determine whether a particular beta receptor subtype is responsible for the development of upregulation as a function of age, we studied the ratio of beta 1- to beta 2-adrenergic receptors in the membrane preparations from the ventricles of Fischer-344 hearts at 6 and 24 months of age. The animals were injected with 6-hydroxydopamine hydrobromide (6-OHDA) (2 x 50 mg/kg, i.v.) on days 1 and 8 and they were decapitated on day 15. The depletion of norepinephrine in the heart was about 86% in each age group. [125I]iodopindolol (IPIN) was used as the radioligand at the final concentration of 110 pM. Inhibition of specific IPIN binding was studied by adding ICI 89,406 (beta 1-selective antagonist) and ICI 118,551 (beta 2-selective antagonist) at 25 pM to 40 microM. The relative proportions of the beta receptor subtypes were determined using a competition radioligand selective binding and computer modeling technique. The ventricles contained about 67% beta 1 and 33% beta 2 adrenergic receptors in hearts isolated from 6- and 24-month old rats; the ratio remained the same in sympathectomized animals. These data suggest that both subtypes of cardiac adrenergic receptors participate in the response to chemical denervation by 6-OHDA regardless of age. PMID- 2554075 TI - [Beta-endorphin in acute focal cerebral infarct]. AB - Experimental studies in animal models suggest that the endorphin system may be implicated in the pathogenetic mechanism of cerebral ischemic lesions. Naloxone has been shown to possess a beneficial effect on the neurologic deficit associated with cerebral ischemia in animal experiments, probably because of its endorphin antagonist properties. By contrast, the results of clinical trials are contradictory. Moreover, the true significance of high plasma levels of beta endorphin in patients with acute focal cerebral infarct (AFCI) has not yet been elucidated. We have evaluated 23 patients with established AFCI, in whom plasma levels of beta-endorphin and corticotropin (ACTH) were simultaneously measured during the first 48 hours after the onset of the disease. The results were compared with those from a control group. In a subgroup of 9 cases new measurements were made after 7 days. In the patients with AFCI, significantly lower levels of beta-endorphin and ACTH than in the control group were found. One week later, a moderate nonsignificant increase in the plasma level of beta endorphin was found. The localization and estimated size of the infarct area were not relevant. Probably, the plasma levels of beta-endorphin will need to be considered before naloxone therapy is indicated, and only if it is confirmed that the plasma levels of beta-endorphin reflect changes at the cerebral level, as the pathophysiological role of these opioids in AFCI has not yet been established. PMID- 2554076 TI - [Tumor markers and primary cancer of the liver]. PMID- 2554077 TI - [Massive lower digestive hemorrhage secondary to cytomegalovirus colitis in a patient with acquired immunodeficiency syndrome]. PMID- 2554079 TI - [Epstein-Barr virus infections--clinical characteristics and diagnosis]. PMID- 2554078 TI - Alpha-adrenoceptors: a critical review. PMID- 2554080 TI - [The role of the surgical approach in the treatment of microcellular carcinoma of the lung]. AB - Twenty-nine patients with small-cell lung cancer undergoing surgery during a 5 year period at the Department of Thoracic Surgery, School of Medicine, University of Zagreb were reviewed. The radical surgical procedure was performed in 22 patients (73.9%). The median survival of these patients was 26.5 months. Two-year survival was 36.4% (eight of 22 patients) and in patients after radical surgery, the four-year survival rate was 18.2% (four of 22 patients). Three patients were alive at the time when this paper was written (60, 59, and 57 months after the initial diagnosis). It is concluded that the surgical therapy is an important part in the multi-disciplinary approach to the treatment of small-cell lung cancer with limited stage. PMID- 2554081 TI - Membrane fluidity of guinea pig lymphocytes and the dysfunction of the respiratory airway and lymphocyte beta-adrenergic systems of the guinea pig. AB - The beta-adrenergic receptor responsiveness of isolated guinea pig tracheal spirals can be negatively affected by intraperitoneal administration of the Gram negative bacterium Haemophilus influenzae, four days prior to the experiment. The reduction in tracheal relaxation is accompanied by a decrease in beta-adrenergic receptor binding sites in splenic lymphocyte membranes and by a decrease in the fluidity of these membranes. The H. influenzae-induced dysfunction of both the respiratory airway and lymphocyte beta-adrenergic systems can be mimicked by modulating the amount of linoleic acid in the diet. This linoleic acid induced dysfunction of the beta-adrenergic system is also accompanied by a decrease in the plasma membrane fluidity of the splenic lymphocyte membranes of the guinea pigs. The role for plasma membrane fluidity in asthma is discussed in relation to current concepts for atopy. PMID- 2554082 TI - Differentiation of phencyclidine and sigma receptor types affecting the central inspiratory termination mechanism in cat. AB - The effects of 1) the phencyclidine receptor ligand TCP, 2) sigma receptor ligands (+)3-PPP and DTG, and 3) N-methyl-D-aspartate receptor blockers MK-801 and dextrorphan were determined on a brainstem mechanism which controls the termination of the inspiratory phase of the breathing cycle. Inspiratory bursts were recorded from the phrenic nerve in decerebrate paralyzed cats ventilated by means of a phrenic driven servoventilator. The central mechanism which terminates inspiration was tested by withholding lung inflation, thus suppressing the contribution of the vagal feedback from the lungs to inspiratory termination. TCP increased the duration of test inspiration (tTi) by 17% at 0.03 mg/kg and by 14 fold (from 1.6 to 23 s) at 1 mg/kg. With dextrorphan, tTi was significantly increased at 3 mg/kg. In contrast, (+)3-PPP and DTG did not increase tTi at doses up to 10 mg/kg, although MK-801 (0.03 mg/kg), given after the sigma ligands, increased tTi by 59-90%. It is concluded that phencyclidine but not sigma receptor ligands block the central mechanism which terminates inspiration and that the likely site of action is the NMDA receptor complex. PMID- 2554083 TI - Characterization of gamma-aminobutyric acid and dopamine overflow following acute implantation of a microdialysis probe. AB - The present study characterized the voltage and calcium dependence of gamma aminobutyric acid and dopamine overflow after the acute implantation of a microdialysis probe. Probes were implanted in dorsolateral striatum and globus pallidus. Experiments were performed under light halothane anesthesia. Basal, extracellular levels of GABA were not affected by tetrodotoxin (TTX) and were increased to 140 percent of basal values by calcium free Ringer. Basal, extracellular levels of dopamine were reduced to 14 percent of basal values by the addition of TTX and to 30 percent of basal values by the removal of calcium from the Ringer solution. The results suggest that in this in vivo preparation basal extracellular dopamine is largely of vesicular origin while GABA is not. PMID- 2554084 TI - A hybridoma producing human monoclonal antibody specific for glycoprotein 120 kDa of human immunodeficiency virus (HIV-1. AB - A stable hybridoma producing anti-HIV human monoclonal antibody (HMCA) was generated by fusing CD3-depleted human splenic lymphocytes from an HIV sero positive donor with the mouse myeloma cell line P3x63AgU1. The resultant hybridoma has been secreting IgG1, lambda chain for over nine months at a rate of 2.5 micrograms/10(6)cells/day. The HMCA shows specific reactivity in ELISA using HIV-infected cell lysates. Immunofluorescence tests have indicated that this HMCA binds specifically to the surface of H9 and C3 HIV/HTLVIIIb infected cells, HIV/N1T infected CEM cells and to MoT cells infected with an HIV clinical isolate. Western blotting revealed recognition of glycoproteins 120 and 160 kDa of HIV by the HMCA. Although this HMCA demonstrated no neutralizing activity, the production of an anti-HIV HMCA specific for glycoprotein 120 kDa indicates the possibility that a neutralizing HMCA can be developed as further fusions with lymph nodes and spleens from HIV positive donors are performed. PMID- 2554085 TI - Two-exponential analysis of spin-spin proton relaxation times in MR imaging using surface coils. AB - Proton relaxation time measurements were performed on a standard whole body MR imager operating at 1.5 T using a conventional surface coil of the manufacturer. A combined CP/CPMG multiecho, multislice sequence was used for the T1 and T2 relaxation time measurements. Two repetition times of 2000 ms (30 echoes) and 600 ms (2 echoes) with 180 degrees-pulse intervals of 2 tau = 22 ms were interleaved in this sequence. A two-exponential T2 analysis of each pixel of the spin-echo images was computed in a case of an acoustic neurinoma. The two-exponential images show a "short" component (T2S) due to white and gray matter and a "long" component (T2S) due to the cerebrospinal fluid. In the fatty tissue two components with T2S = 35 +/- 3 ms and T2L = 164 +/- 7 ms were measured. Comparing with Gd-DTPA imaging the relaxation time images show a clear differentiation of vital tumor tissue and cerebrospinal fluid. PMID- 2554086 TI - [Radiodiagnosis of the reparative osteogenesis during the treatment of comminuted fractures of the humerus]. AB - Roentgenoradiological control of a reparative process after diaphysial comminuted shoulder fractures was exercised at all stages of the treatment after G. A. Ilizarov's method. Osteoscanning permitted the assessment of the viability of bone splinters on the first days after trauma and quantitative assessment of the activity of osteogenesis. A study of the optical density of radiograms provided an opportunity for evaluation of mineral content in regenerating bones, their structure only at late stages of treatment when it was rather high. PMID- 2554087 TI - [Effect of smoking on the hypophyseo-adrenal axis]. AB - Plasma concentrations of adrenocorticotrophin (ACTH) and cortisol were measured during insulin-induced hypoglycemia and lysin-8-vasopressin (LVP) test in 60 healthy subjects, non-smokers and habitually smokers of 10 or more cigarettes/24 hours. A marked and statistically significant rise of both hormones was found in non-smoker subjects, whereas smokers showed poor and not significant modifications. These results suggest that continuous chronic inhalation of nicotine may act as a powerful stimulus on the hypothalamo-hypophyseal structures that control the hypothalamic CRF and/or ACTH production and release. Central nervous mechanisms of hormonal regulation may become less sensitive and efficient when an acute rise of ACTH is required, as during stimulating tests. Our investigation confirms that cigarette smoking is heavily responsible of endocrine disorders, in particular of hypophyseal-adrenal axis. PMID- 2554088 TI - [Use of catheterization of the inferior petrosal sinus in the diagnosis of Cushing's disease]. AB - Previous reports have emphasized the value of bilateral and simultaneous catheterization of the inferior petrosal sinus for the measurement of both basal and oCRH (ovine Corticotropin Releasing Hormone) stimulated ACTH levels to determine the site of the microadenoma in the pituitary and for the differential diagnosis of Cushing's disease. This method is mainly employed in those patients whose hormonal studies are ambiguous and whose CT-scans and NMR (Nuclear-Magnetic Resonance) results yield inconclusive or negative findings. Ten patients were studied: 9 were under evaluation for Cushing's disease and one was a "relapse" (4 yrs after transsphenoidal microadenomectomy). In all patients, except two who are awaiting surgery and one who was a "relapse", surgical findings were consistent with the ipsilateral hypersecretion (one had a central microadenoma). These results confirm that bilateral and simultaneous catheterization of the inferior petrosal sinus associated with a oCRH stimulus could be of great help in localizing the site of the adenoma and therefore improve the results of surgery. Moreover, this methodology may be of great value in diagnosing ectopic secretion. PMID- 2554089 TI - The Epstein-Barr virus and nasopharyngeal carcinoma. PMID- 2554090 TI - myc-related proteins and DNA sequences in Trypanosoma brucei. AB - The cAMP content of Trypanosoma brucei increases in parallel with ascending mammalian parasitemia to very high levels just before differentiation of the long slender to the short-stumpy bloodstream form. Because expression of myc oncogenes is required for vertebrate cells to interpret proliferation signals and declines in response to cAMP mediated differentiation, we investigated whether T. brucei also harbored myc-like proteins and genes. Accordingly, we probed lysates of long slenders, short-stumpies and procyclics (insect midgut stage) with antibody to myc proteins and also hybridized myc gene family sequences to procyclic DNA. We found that antibody to myc-family proteins of mammals reacts with 40 kDa and 55 kDa proteins in all three life cycle stages, and that procyclic DNA contains three EcoRI fragments that are homologous to a v-myc probe. One of these fragments also hybridizes to a synthetic 25-mer oligonucleotide deduced from a consensus sequence in the second exon of the myc family and expresses a 3.2 kb mRNA transcript in Northern blots of procyclic RNA. The conservation of myc family homologous across the broad phylogenetic gap between mammals and trypanosomes illustrates ancient evolutionary relationships and raises the possibility of stage-specific expression of myc genes during the life cycle of T. brucei. PMID- 2554091 TI - Exogenous immunosuppression with methylprednisolone does not reactivate a type D retrovirus in rhesus monkeys. AB - To determine whether immunosuppression would result in retrovirus expression in previously infected rhesus monkeys, chronic high dose methyl prednisolone therapy was administered to two groups of animals for 4 weeks. One group was antibody positive for a type D retrovirus, designated Type D/3/wisc. The second group of animals had no known exposure to Type D/3/wisc and was antibody negative to this virus. The monkeys were evaluated for immunosuppression and retrovirus re expression following the corticosteroid therapy. Although this treatment induced a marked cellular immunosuppression in all animals, as measured by in vitro assays, in none of the animals was retrovirus viremia or retrovirus-associated disease detected. PMID- 2554092 TI - Receptors for and bioresponses to 1,25-dihydroxyvitamin D in a human colon carcinoma cell line (HT-29). AB - Human colon carcinoma (HT-29) cells were examined for their capacity to bind and respond to 1,25-dihydroxycholecalciferol [1,25-(OH)2D3]. These cells are known to differentiate and increase their population doubling time when galactose is substituted for glucose in their media. High-affinity and specific binding of 1,25-(OH)2[3H]D3 was observed in extracts of these cells grown in glucose. The binder sedimented in sucrose gradients and eluted from DEAE-cellulose columns in a manner indistinguishable from rabbit intestinal 1,25-(OH)2D3-receptor. Smaller amounts of this binder were seen in HT-29 cells grown in galactose. Both glucose fed and galactose-fed cells exhibited a dose-dependent decrease in growth rate on exposure to 10(-12) to 10(-6) mol/L 1,25-(OH)2D3. Ultrastructural examination of galactose-fed and glucose + 1,25-(OH)2D3-treated cells showed enterocytic differentiation and features that were not distinguishable between these groups. Sucrase activity was higher in galactose-fed cells and did not change with 1,25 (OH)2D3 treatment. However, the lower sucrase activity in glucose-fed cells increased after exposure to 10(-8) mol/L 1,25-(OH)2D3. These results indicate receptor content and bioresponsivity to 1,25-(OH)2D3 in a human enterocytic cell line, suggesting that it will be a useful model for the study of the mechanisms of action of this sterol. PMID- 2554093 TI - Dietary fat-mediated changes in hepatic apoprotein B/E receptor in the guinea pig: effect of polyunsaturated, monounsaturated, and saturated fat. AB - These studies investigated the effects of dietary fat quality on guinea pig plasma total and low-density lipoprotein (LDL) cholesterol and triglyceride levels; LDL composition, density, and binding affinity to hepatic apolipoprotein (apo) B/E receptors; and hepatic apo B/E receptor levels. Animals were fed semi synthetic diets containing 7.5% (weight per weight) fat--either corn oil (CO), olive oil (OL), or lard. Plasma cholesterol levels were significantly lower on the CO diet compared with the OL and lard diets. The isolated LDL had mean peak densities that ranged from 1.071 for LDL from lard-fed animals to 1.075 for LDL from CO- and OL-fed animals. The cholesterol ester to protein ratio of the LDL particle decreased in CO-fed guinea pigs. Binding studies showed that these compositional changes of the LDL had no effect on the binding affinity of the particles to a standardized hepatic membrane preparation. Membrane phospholipid fatty acid compositions were significantly different among the three dietary fat groups. When hepatic membranes were incubated with 125I-labeled LDL, the receptor mediated binding of LDL to membranes from CO-fed animals increased 1.5-fold compared with binding to membranes from OL- and lard-fed guinea pigs. Scatchard plots indicated an increase of 50% in receptor number in membranes of animals fed the CO diet, whereas the affinity of the hepatic apo B/E receptor for LDL (Kd) was virtually identical for all membranes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554094 TI - Interaction of very-low-density lipoprotein isolated from type I (insulin dependent) diabetic subjects with human monocyte-derived macrophages. AB - Very-low-density lipoproteins (VLDL) (density less than 1.006 g/mL) were isolated from type I (insulin-dependent) diabetic patients in good to fair glycemic control and from age-, sex-, and race-matched, nondiabetic, control subjects. VLDL were incubated with human, monocyte-derived macrophages obtained from nondiabetic donors, and the rates of cellular cholesteryl ester synthesis and cholesterol accumulation were determined. VLDL isolated from diabetic patients stimulated significantly more cholesteryl ester synthesis than did VLDL isolated from control subjects (4.04 +/- 1.01 v 1.99 +/- 0.39 nmol 14C-cholesteryl oleate synthesized/mg cell protein/20 h; mean +/- SEM, P less than .05). The stimulation of cholesteryl ester synthesis in macrophages incubated with VLDL isolated from diabetic patients was paralleled by a significant increase in intracellular cholesteryl ester accumulation (P less than .05). The increase in cholesteryl ester synthesis and accumulation in macrophages were mediated by a significant increase in the receptor mediated, high affinity degradation (2.55 +/- 0.23 v 2.12 +/- 0.20 micrograms degraded/mg cell protein/20 h) and accumulation (283 +/- 35 v 242 +/- 33 ng/mg cell protein/20 h) of 125I-VLDL isolated from diabetic patients compared with VLDL from control subjects. To determine if changes in VLDL apoprotein composition were responsible for the observed changes in cellular rates of cholesteryl ester synthesis and accumulation, we also examined the apoprotein composition of the VLDL from both groups. There were no significant differences between the apoproteins B, E, and C content of VLDL from both groups. We also determined the chemical composition of VLDL isolated from both groups of subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554095 TI - The pharmacology of dichloroacetate. AB - Dichloroacetate (DCA) exerts multiple effects on pathways of intermediary metabolism. It stimulates peripheral glucose utilization and inhibits gluconeogeneis, thereby reducing hyperglycemia in animals and humans with diabetes mellitus. It inhibits lipogenesis and cholesterolgenesis, thereby decreasing circulating lipid and lipoprotein levels in short-term studies in patients with acquired or hereditary disorders of lipoprotein metabolism. By stimulating the activity of pyruvate dehydrogenase, DCA facilitates oxidation of lactate and decreases morbidity in acquired and congenital forms of lactic acidosis. The drug improves cardiac output and left ventricular mechanical efficiency under conditions of myocardial ischemia or failure, probably by facilitating myocardial metabolism of carbohydrate and lactate as opposed to fat. DCA may also enhance regional lactate removal and restoration of brain function in experimental states of cerebral ischemia. DCA appears to inhibit its own metabolism, which may influence the duration of its pharmacologic actions and lead to toxicity. DCA can cause a reversible peripheral neuropathy that may be related to thiamine deficiency and may be ameliorated or prevented with thiamine supplementation. Other toxic effects of DCA may be species-specific and reflect marked interspecies variation in pharmacokinetics. Despite its potential toxicity and limited clinical experience, DCA and its derivatives may prove to be useful in probing regulatory aspects of intermediary metabolism and in the acute or chronic treatment of several metabolic disorders. PMID- 2554096 TI - Effect of a hyperlipidic diet on lipid composition, fluidity, and (Na+-K+)ATPase activity of rat erythrocyte membranes. AB - Feeding rats a hyperlipidic diet in which animals were offered daily a variety of high-energy food resulted in a significant increase of serum free fatty acids and a decrease of phospholipids with respect to controls. On the contrary, there were no significant differences in erythrocyte membrane total lipid composition between the two groups. Erythrocyte membranes showed a significant decrease in saturated fatty acid content and a significant increase in (n-6) polyunsaturated fatty acid content; (n-3) polyunsaturated fatty acids significantly decreased. Membrane fluidity, investigated by fluorescence polarization of diphenylhexatriene, significantly increased in the erythrocyte membranes of the experimental group. These results seem compatible with the decreased saturated/unsaturated fatty acid ratio. A significant decrease of (Na+-K+)ATPase activity occurred in erythrocyte membranes of the experimental group rats with respect to the controls. PMID- 2554097 TI - Muscarinic-agonist and guanine nucleotide stimulation of myo-inositol trisphosphate formation in membranes isolated from bovine iris sphincter smooth muscle: effects of short-term cholinergic desensitization. AB - The effect of short-term cholinergic desensitization on muscarinic acetylcholine receptor (mAChR)-mediated activation of phospholipase C was investigated in membranes isolated from the bovine iris sphincter smooth muscle. Membranes prepared from normal or desensitized muscles, prelabeled with either [3H]myo inositol or 32P from [gamma-32P]ATP, were incubated with a hydrolysis-resistant analogue of GTP, GTP gamma S, or GTP gamma S plus carbachol (CCh), and the production of [3H]myo-inositol 1,4,5-trisphosphate (IP3) and the breakdown of polyphosphoinositides were assessed. In normal membranes, GTP (greater than or equal to 1 mM), GTP gamma S (greater than 10 microM) and GTP gamma S (1 microM) plus CCh (10 microM), but not GDP or GDP beta S, increased phosphatidylinositol 4,5-bisphosphate (PIP2) hydrolysis and IP3 production. GTP gamma S increased IP3 accumulation in a time- and dose-dependent manner, and CCh, which had no effect on phospholipase C activity in the absence of GTP gamma S, potentiated the effects of GTP gamma S. The effect of CCh plus GTP gamma S on IP3 production was inhibited by atropine, had an absolute requirement for nM amounts of Ca2+ and was not affected by pertussis toxin. At higher concentrations (greater than 1 microM), Ca2+ alone induced PIP2 hydrolysis. Short-term exposure (less than 60 min) of the muscle to CCh (100 microM) did not affect the total number (Bmax) of mAChRs nor their affinity (KD) for [3H]-N-methylscopolamine. Desensitization did, however, result in: (1) a loss of the CCh-high affinity binding state of the sphincter mAChRs in a manner analogous to that produced by GTP gamma S; (2) a loss of the ability of GTP gamma S to affect CCh binding to the receptors; and (3) an attenuation of the GTP gamma S plus CCh-stimulated PIP2 hydrolysis. In conclusion, the data presented suggest that, in the iris smooth muscle, G proteins are involved in the coupling of mAChRs to phospholipase C and that short term cholinergic desensitization results in (1) the uncoupling of the receptor-G protein complex and (2) the attenuation of mAChR-activation of phospholipase C. PMID- 2554098 TI - Cell fusion induced by herpes simplex is inhibited by hen egg-white lysozyme. AB - The formation of syncytia in cell monolayers infected with a macroplaque strain (MP) of herpes simplex virus was found to be inhibited by hen egg-white lysozyme. Inhibition was roughly proportional to the enzyme concentration. The virus titres in supernatant fluids of lysozyme-treated cultures were also reduced compared with untreated cultures. Control experiments excluded the possibility that lysozyme altered the virus viability and infectivity or impaired cell growth. Since lysozyme is a cationic protein, further experiments were performed in order to discover whether its antisyncytiogenic effect depended on its enzymatic activity or on its positive charge. Inhibition of the MP-induced polycaryocytosis was found to be caused by heat-inactivated lysozyme and three chemically-modified lysozymes with a higher positive charge (one retaining and two lacking enzymatic activity). PMID- 2554099 TI - Hand-to-hand transmission of herpes simplex virus type 1. AB - Droplets of tissue culture fluid containing herpes simplex virus type 1 were placed on the palm of the hand. Each 0.01 ml droplet was taken from a stock virus suspension with a titre of 10(7.5) TCID50/0.1 ml. At 0, 15, 30, 60 and 120 min a droplet was firmly touched with the middle finger of the right hand, after which, attempts were made to recover virus from the finger. At 0 min, when the virus containing droplet was in a liquid state, there was a 100% rate of virus recovery. By 15 min the droplets had dried out, and after touching dried out droplets there was a 40% virus recovery rate, even though experimental procedures demonstrated that infectious virus was present in the dried out droplets at all test times. If the finger was moistened with tap water or saliva, there was a 100% recovery rate of virus after touching dried out droplets, as well as after touching droplets in a liquid state. PMID- 2554100 TI - Ifosfamide and mesna. PMID- 2554101 TI - [Value of diagnostic localization of insulinoma]. AB - The value of diagnostic localization of insulinoma The value of diagnostic localization statements are contradictory. Basing on our own patient material (n = 41) the preoperative localization of an insulinoma was correct with sonography in 57.7%, with computed tomography in 21.4%, with computed tomography with bolus injection of contrast medium in 73.3%, with angiography 63.9% and with percutaneous transhepatic portal vein catheterisation with selective test of hormones (PTP) in 76.9%. Intraoperative 38 of 41 insulinomas were palpable and twelve of 16 insulinomas were seen during intraoperative sonography. Although we palpate more than 90% of all insulinomas we support a preoperative diagnostic localization for easier intraoperative palpation. PMID- 2554102 TI - [Unilateral pulsating tinnitus: paraganglioma of the glomus jugulare with predominantly vascular extension]. PMID- 2554103 TI - Effects of selective oxidation of 1-34 bovine parathyroid hormone on its renal actions in the rabbit. AB - Oxidation of both of the methionine residues (positions 8 and 18) in parathyroid hormone (PTH) eliminates many of its biological effects. The present studies were performed to examine the actions of 1,34 bovine PTH and 1-34 bovine PTH oxidized selectively at Met 8, at Met 18, and at both sites on renal electrolyte handling and on adenylate cyclase (AC) stimulation. In clearance studies in anesthetized rabbits, PTH caused a phosphaturia and an anticalciuria. PTH also stimulated renal proximal tubular AC in vitro and increased renal cortical cAMP content in vivo. PTH oxidized at Met 18 was anticalciuric, but not phosphaturic, stimulated renal AC and increased cortical cAMP content. PTH oxidized at Met 8 also produced an anticalciuria without a phosphaturia, but only weakly stimulated AC and did not alter cortical cAMP content. PTH oxidized at both Met 8 and Met 18 was phosphaturic but not anticalciuric, was a weak agonist for AC and decreased cortical cAMP content. In the isolated perfused rabbit proximal straight tubule, PTH inhibited fluid and phosphate transport, whereas the doubly oxidized peptide was inactive. The data are consistent with the possibility that the effects of PTH on renal tubular phosphorus transport are mediated by more than one mechanism and are, in part, independent of the cAMP messenger system. PMID- 2554105 TI - Sensitivity: the special role in oncology. PMID- 2554104 TI - Plasma ouabain-like activity in essential hypertensive patients and in subjects with primary aldosteronism. AB - Plasma from 37 essential hypertensive patients, from 11 subjects with primary aldosteronism and from 23 normotensive subjects was tested for ouabain-like activity. Despite a very substantial overlap, hypertensive patients (both essential and secondary) showed significantly higher levels of a ouabain-like plasma factor compared to normotensive controls. No substantial differences could be detected, however, between the two forms of hypertension; in particular, no significant changes were observed in the low-PRA subgroup. Our results are hardly compatible with the hypothesis that this substance may be of crucial importance in the development either of essential hypertension or of primary aldosteronism. PMID- 2554107 TI - Multimodality therapy for lung cancer. PMID- 2554106 TI - More than just junk food. PMID- 2554108 TI - [Nourishment of the hepatocellular carcinoma via portal blood flow--from the analysis of the tumor cell kinetics using 5-bromo-2'-deoxyuridine: preliminary report]. PMID- 2554109 TI - Multiple affinity binding states of the sigma receptor: effect of GTP-binding protein-modifying agents. AB - The sigma receptor, which is labeled with (+)-[3H]3-(3-hydroxyphenyl)-N- 1 (propyl)piperidine [(+)-[3H]3-PPP], is a site that binds several psychotomimetic opiate benzomorphans and certain antipsychotics, such as haloperidol. In order to elucidate the mechanisms involved in sigma receptor ligand binding, equilibrium binding analysis and kinetics of association and dissociation of the relatively selective sigma receptor ligand (+)-[3H]3-PPP were determined in rat brain membranes in the absence and presence of 5'-guanylylimidodiphosphate [Gpp(NH)p]. In the absence of Gpp(NH)p, (+)-3-PPP, cyclazocine, pentazocine, and (+)-SKF 10047 bind to high and low affinity sites (KH = 1.3-7.5 nM; KL = 84-500 nM), as determined by computer assisted analysis of the inhibition of (+)-[3H]3-PPP binding by the sigma ligands. The antipsychotics haloperidol and chlorpromazine inhibit (+)-[3H]3-PPP binding in a manner indicating interaction with a single state of the receptor. Gpp(NH)p (0.1 mM) abolished the high affinity binding component of the sigma agonist-like compounds tested but had no effect on the affinities of the antipsychotics for the receptor. Gpp(NH)p decreased the association rate of (+)-[3H]3-PPP binding 5-fold and also converted the biexponential dissociation kinetics of the ligand, observed in the absence of Gpp(NH)p, to a rapid monophasic dissociation process. Pretreatment of membranes with N-ethylmaleimide and pertussis toxin inhibited (+)-[3H]3-PPP binding and abolished the effect of Gpp(NH)p on the sigma ligand binding. These findings indicate of the sigma receptor is capable of existing in two discrete states, having high and low affinity for sigma agonist-like drugs. The regulation of the high affinity binding state by GTP-binding protein-modifying agents suggests its coupling to GTP-binding protein(s). PMID- 2554110 TI - Ca2+-mediated neuronal death in rat brain neuronal cultures by veratridine: protection by flunarizine. AB - Neuronal cell degeneration was studied in vitro in primary rat brain neuronal cultures grown in serum-free, chemically defined, CDM R12 medium, by measuring lactate dehydrogenase (LDH) released in the culture medium. A Ca2+-dependent neuronal cell degeneration was observed after prolonged and transient exposure 30 microM veratridine. The release of LDH occurred gradually and could be completely prevented by 2 mM ethylene glycol bis (beta-aminoethyl ether)-N,N,N',N' tetraacetic acid, 0.1 microM tetrodotoxin, and 1 microM flunarizine. Flunarizine was without effect on neuronal cell loss induced by 1 mM glutamate, 1 mM kainic acid, and 5 mM KCN. The lack of effect on neurotoxicity induced by 1 mM glutamate differentiates flunarizine from N-methyl-D-aspartate antagonists such as MK-801. The latter protected at nanomolar concentrations against glutamate-induced neuronal cell death but had a maximal effect only at 0.1 mM on the veratridine induced released LDH. It is suggested that, besides the excitatory amino acid receptor pathway, prolonged opening of the veratridine-sensitive Na+ channel can be neurotoxic. The latter can be prevented by flunarizine. The role of Na+ channel blockers as therapeutic agents in cerebral ischemia is discussed. PMID- 2554111 TI - Structural requirements for activation of the glycine coagonist site of N-methyl D-aspartate receptors expressed in Xenopus oocytes. AB - Five structural features important for activation of the glycine recognition site on N-methyl-D-aspartate (NMDA) receptors were identified by structure-activity studies of more than 60 glycine analogues in voltage-clamped Xenopus oocytes injected with rat brain mRNA. First, sterically unhindered and ionized carboxyl and amino termini were essential for action at this site. Second, an increase in the interterminal separation by greater than one carbon (e.g., beta-alanine) markedly attenuated activity at this site. Third, activity at the glycine site was stereoselective. The D-isomers of alanine and serine were approximately 20 and 30 times more potent than the L-isomers. Fourth, only small sterically unobtrusive substitutions at the alpha-carbon could be tolerated. alpha-Methyl (D alanine) and alpha-cyclopropyl (1-amino-cyclopropane carboxylic acid) (ACC) substitutions were effective as agonists but most larger aliphatic and aromatic alpha-carbon substitutions were simply inactive. Glycine, D-alanine, and ACC probably have only a two-point attachment to the receptor. Finally the alpha carbon substituent of D-serine is envisioned as binding to a third site on the receptor probably via hydrogen bonding of the omega-terminal hydroxyl group. Thus, serine, an hydroxymethyl substitution of glycine, permitted activation of NMDA receptor-mediated currents, whereas isosteric substitutions incapable of hydrogen bonding (e.g., 2-aminobutyric acid) were inactive. Additionally, the position and size of the hydroxyl-containing group is critical for agonist action; D-threonine, DL-homoserine, and hydroxyphenolic substitutions at the alpha-carbon were all inactive. Halogenated analogs of a size comparable to D serine but capable only of proton acceptance at the omega-terminus (beta-fluoro-D alanine and beta-chloro-D-alanine) possessed agonist action, whereas an analog capable of only proton donation (1,2-diaminopropionic acid) was inactive. Full concentration-response curves were constructed for those analogs displaying greater than 25% of the effect of glycine when tested at 3 microM. With the exception of (R)-(+)cycloserine and beta-fluoro-D-alanine, all compounds were nearly full agonists and had Hill coefficients not significantly different from unity. The order of relative potency of the active analogs was ACC greater than glycine greater than D-serine greater than D-alanine greater than beta-fluoro-D alanine greater than (R)-(+)-cycloserine greater than L-serine greater than L alanine. Molecular modelling of a series of active and inactive analogs with close structural relation to glycine was undertaken. These results were complementary to those data obtained from the electrophysiological investigation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2554112 TI - Effects of polyamines on the binding of [3H]MK-801 to the N-methyl-D-aspartate receptor: pharmacological evidence for the existence of a polyamine recognition site. AB - A heat-stable factor of low molecular weight that increases the binding of [3H]MK 801 to rat brain membranes in the presence of maximally effective concentrations of L-glutamate and glycine was purified from bovine brain by reverse phase and ion-exchange high pressure liquid chromatography. The stimulatory activity was due to the presence of spermidine in the active fractions. Polyamines including spermine and spermidine are found in high concentrations in mammalian tissue. These compounds increase the affinity of N-methyl-D-aspartate (NMDA) receptors for [3H]MK-801 when assays are carried out in the presence of 100 microM L glutamate and 100 microM glycine. At concentrations of 1 to 300 microM, a number of di- and triamines, including NH2(CH2)3NH2, NH2(CH2)3NH(CH2)2NH2, and NH2(CH2)3NH(CH2)3NH2, have partial or full agonist-like activity similar to that of spermidine. Other polyamines, including putrescine, cadaverine, NH2(CH2)2NH(CH2)2NH2, and CH3NH(CH2)3NHCH3, at concentrations of 1 to 100 microM, inhibited the binding of [3H]MK-801 in the presence of spermine, L-glutamate, and glycine but not in the presence of only L-glutamate and glycine. It is concluded that these compounds are selective antagonists of the effects of spermine at the NMDA receptor. These results suggest that there may be a polyamine recognition site on the NMDA receptor complex. PMID- 2554113 TI - Molecular determinants of benzodiazepine receptor affinities and anticonvulsant activities. AB - In vivo convulsant activities profiles and receptor binding studies together with the techniques of theoretical chemistry were used to characterize 15 compounds, from five different chemical families, known to bind to the BDZ receptor. The experimental goals of this study were to determine the affinity of these analogs for this receptor, the effect of gamma-aminobutyric acid on the affinity, and, in a self-consistent manner, the nature of the activity, agonist (anticonvulsant), antagonist, or inverse agonist (proconvulsant, convulsant), elicited by binding to this receptor. To these ends, in vivo studies were made to determine the proconvulsant, convulsant, and anticonvulsant activities and antagonism to anticonvulsant activities of the 15 analogs. Their receptor affinities at 25 degrees were also determined by competitive inhibition of [3H] flunitrazepam and [3H]Ro 15-1788 in the absence and presence of gamma-aminobutyric acid. The goal of the theoretical studies was to identify and calculate molecular properties that modulate these affinities and types of activities and from them to develop a model of receptor recognition and activation that could consistently explain observed behavior and predict new results. Thus, molecular orbital calculations were carried out for all analogs, using semiempirical quantum mechanical methods. In addition to the optimization of structures, a number of electronic properties, such as polarizations, partition coefficients, and proton and electron affinities were computed and examined for their ability to modulate relative affinities and modes of activation of the receptor. From these studies, a model for receptor recognition involving two anchoring hydrogen bond-acceptor sites and for activation involving interaction of the most lipophilic aromatic region of each compound with the receptor was developed, which could systematically account for the three different types of behavior, agonist, antagonist, and inverse agonist, observed for these analogs. Electronic rather than structural properties were found to be the principal modulator of both recognition and activation. A possible mechanism of agonist activation of the receptor involving electron transfer to the agonist, as well as a possible induced conformational change in the receptor, is also suggested by these results. Finally, by complementarity, some steric and electronic characteristics of the receptor binding site could be deduced. PMID- 2554114 TI - Comparison of location and binding for the positively charged 1,4-dihydropyridine calcium channel antagonist amlodipine with uncharged drugs of this class in cardiac membranes. AB - The distinctive pharmacokinetic and pharmacodynamic activity of amlodipine, including long onset and duration of activity as a calcium channel antagonist, may be related to its interactions with membranes. We have used X-ray crystallography and small-angle X-ray scattering to examine and compare the crystal structure of amlodipine and its location in cardiac sarcolemmal lipid bilayers with that of uncharged dihydropyridines (DHPs) such as nimodipine. Crystallographic analysis demonstrated that the DHP ring of amlodipine is considerably more planar than that of nimodipine, that amlodipine has a greater torsion angle between the DHP and aryl rings, and that the protonated amino group extends away from the DHP ring structure. Despite the positive charge of amlodipine at physiological pH, membrane electron density profile structures showed amlodipine to have a time-averaged location near the hydrocarbon core/water interface similar to that observed for several uncharged DHPs. However, unlike uncharged DHPs, this location is consistent with an ionic interaction between the protonated amino function of amlodipine and the negatively charged phospholipid headgroup region, in addition to a hydrophobic interaction with the fatty acyl chain region near the glycerol backbone similar to other DHPs. This location may also provide an appropriate conformation and orientation for amlodipine binding to its receptor site at this depth in the membrane. Finally, we have measured the nonspecific partitioning of amlodipine into native sarcoplasmic reticulum membranes from rabbit skeletal muscle and compared these data with those for the uncharged DHPs. The partition coefficient into light sarcoplasmic reticulum for amlodipine was higher than that observed for most uncharged DHPs and rates of incorporation of amlodipine into membranes were very high, as with other DHPs, whereas the "washout time" of amlodipine from these membranes was longer by over 1 order of magnitude. These data suggest differences in membrane interactions for amlodipine, compared with uncharged DHPs, that may be correlated with its novel pharmacodynamic and pharmacokinetic profile. PMID- 2554115 TI - Altered patterns of agonist-stimulated cAMP accumulation in cells expressing mutant beta 2-adrenergic receptors lacking phosphorylation sites. AB - As with many other receptor-effector systems, the responsiveness of the beta adrenergic receptor (beta AR)/adenylyl cyclase system undergoes desensitization upon agonist exposure. Phosphorylations of the receptor by the cAMP-dependent protein kinase (protein kinase A) and the beta AR kinase appear to play roles in such desensitization phenomena, but the functional significance of the receptor phosphorylation in intact cells has not been previously assessed. In this study, we constructed and expressed in a mammalian fibroblast line the normal (wild type) human beta 2 AR and mutant forms of the receptor that lack the putative phosphorylation sites for these two protein kinases. The two consensus sequences for phosphorylation by protein kinase A were altered by changing serines 261, 262 and 345, 346 to alanines. In another mutant, the 11 serines and threonines at the carboxy terminus of the protein that constitute the putative beta AR kinase phosphorylation sites were changed to alanines or glycines. The mutated receptors did not differ from the wild type in their affinities for agonists or antagonists or in their ability to mediate agonist stimulation of adenylyl cyclase. Moreover, their levels of expression in the cultured cells were the same. When stimulated with the beta AR agonist isoproterenol, cells bearing either the wild type or mutant receptors generated cAMP at essentially identical rates for the first 2 min. Cells bearing wild type receptors then showed a rapid desensitization characterized by a markedly diminished rate of cAMP production after the first few minutes of stimulation. However, cells bearing either of the mutated forms of the receptor showed much less desensitization and continued to generate cAMP at a rate 3-4 times greater than that observed in cells expressing the wild type receptor. In contrast, intact cell cAMP levels stimulated by prostaglandin E1 and forskolin were not different between cells bearing wild type or mutant beta AR. These results suggest an important physiological role for phosphorylation of the beta AR in regulating rapid agonist-induced desensitization in intact cells. PMID- 2554116 TI - Direct inhibition of inositol-1,4,5-trisphosphate-induced Ca2+ release from brain microsomes by K+ channel blockers. AB - Tetraethylammonium and 9-tetraethylammonium have previously been reported to inhibit inositol-1,4,5-trisphosphate (IP3)-induced Ca2+ release from brain microsomes, purportedly by blocking potassium channels [Biochem. J. 258:617-620 (1988)]. The effects of these and other K+ channel blockers have been studied here in greater detail using a spectrophotometric assay for Ca2+ movements into and out of canine brain microsomes. IP3-induced Ca2+ release was inhibited by substitution of K+ in the medium with nominally impermeant cations or by addition of most of the K+ channel blockers tested. Nevertheless, addition of valinomycin to the medium (to provide an alternative pathway for counter-ion K+ movements) failed to alleviate the inhibition of IP3-induced Ca2+ release caused by K+ channel blockers. To determine whether these substances act by inhibition of IP3 binding or by direct interaction with the Ca2+ channel of the internal store that promotes IP3-induced Ca2+ release, their effect on [3H]IP3 binding was investigated. None of the K+ channel blockers tested inhibited [3H]IP3 binding. Nearly all the K+ channel blockers appear to interact directly with a Ca2+ channel of the intracellular stores or perhaps interfere with its coupling to the IP3 receptor. Because of their multiplicity of actions, these substances cannot be presumed to be either selective K+ channel blockers or selective inhibitors of IP3-induced Ca2+ release from internal stores. Three of them were even found to partially inhibit valinomycin-stimulated 86Rb uptake into liposomes. PMID- 2554117 TI - Pharmacologic differentiation between inositol-1,4,5-trisphosphate-induced Ca2+ release and Ca2+- or caffeine-induced Ca2+ release from intracellular membrane systems. AB - Various known Ca2+ channel blockers and intracellular Ca2+ antagonists have been tested for effects of inositol-1,4,5-trisphosphate (IP3)-induced Ca2+ release from isolated canine brain microsomes. In agreement with previous reports, heparin, p-chloromercuribenzoic acid, W-7, cinnarizine, flunarizine, certain local anesthetics, La3+, and Ca2+ inhibit the release of Ca2+ induced by addition of IP3. In addition, we report here pronounced inhibition of IP3-induced Ca2+ release by low levels of Cd2+, by relatively high concentrations of TMB-8, and by phytic acid. In contrast, a number of blockers of other Ca2+ channels (nifedipine, verapamil, dantrolene, dithiothreitol, and ruthenium red) have relatively little or no effect on IP3-induced Ca2+ release from brain microsomes. The relative ineffectiveness of substances that inhibit Ca2+- or caffeine-induced Ca2+ release from skeletal muscle sarcoplasmic reticulum suggests that release of Ca2+ from caffeine- and IP3-sensitive neuronal Ca2+ stores is likely to be mediated by different channels. Further evidence that different channels are involved is presented by way of demonstration of the lack of Ca2+-induced Ca2+ release from these brain microsomes and the lack of effect on sarcoplasmic reticulum caffeine-induced Ca2+ release of certain inhibitors of IP3-induced Ca2+ release used here. Among IP3-induced Ca2+ release blockers, La3+ appeared to be exceptional in its ability to stimulate microsomal Ca2+ uptake sufficiently to attenuate release of Ca2+ induced by IP3. Most blockers of IP3-induced Ca2+ release appear not to function by way of inhibiting K+ counter-ion movements (valinomycin does not reverse the inhibition) but rather by way of direct interaction with the IP3 receptor or the Ca2+ channel that mediates the IP3 induced Ca2+ release. Inhibition of [3H]IP3 binding to the microsomes by phytic acid, heparin, pyrophosphate, p-chloromercuribenzoic acid, and Ca2+ could be demonstrated but not by the other substances tested. PMID- 2554118 TI - Calcium movements during each heart beat. AB - Transsarcolemmal calcium movements are closely related to force generation in the heart. It is important to understand the transport pathways that control these movements of calcium across the sarcolemmal membrane. In the normal, beating heart, sodium-calcium exchange appears to be an important mechanism for the extrusion of calcium from the cell. The kinetics of this exchange are dependent upon the characteristics of the cell action potential. Calcium efflux via sodium calcium exchange may be sufficient to balance calcium entry through calcium channels during the action potential. PMID- 2554119 TI - The control of calcium influx by cytoplasmic calcium in mammalian heart muscle. AB - The role of Ca2+ in the initiation and maintenance of contraction has been extensively studies. Many of these studies have focused on how Ca2+ influx and efflux affect cytoplasmic Ca2+ (Cai) and, therefore, contraction in cardiac muscle. However, it has recently become apparent that Cai itself may play a major role in the control of Ca2+ influx and efflux from cardiac muscle. Here we review current ideas on the mechanisms underlying Ca2+ homeostasis in cardiac muscle, with specific attention to how Cai may control Ca2+ influx, both under normal and pathological conditions. PMID- 2554120 TI - Excitation-contraction coupling in heart muscle. AB - We have investigated the links between electrical excitation and contraction in mammalian heart muscle. Using isolated single cells from adult rat ventricle, a whole-cell voltage-clamp technique and quantitative fluorescence microscopy, we have measured simultaneously calcium current (ICa) and [Ca2+]i (with fura-2). We find that the voltage-dependence of ICa and the [Ca2+]i-transient and the dependence of [Ca2+]i-transient on depolarization-duration cannot both be readily explained by a simple calcium-induced Ca-release ('CICR') mechanism. Additionally, we find that when [Ca2+]i and [Na+]i are at their diastolic levels, activation of the Na-Ca exchange mechanism by depolarization does not measurably trigger the release of Ca2+i. Finally, measuring ICa in adult and neonatal rat heart cells and using the alkaloid ryanodine, we have carried out complementary experiments. These experiments show that there may be an action of ryanodine on ICa that is independent of [Ca2+]i and independent of a direct action of the alkaloid on the calcium channel itself. Along with experiments of others showing that ryanodine binds to the sarcoplasmic reticulum calcium-release channel/spanning protein complex, our data suggests a model to explain our findings. The model links the calcium channels responsible for ICa to the sarcoplasmic reticulum by means of one or more of the spanning protein(s). Information from the calcium channel can be communitated to the sarcoplasmic reticulum by this route and, presumably, information can move in the opposite direction from the sarcoplasmic reticulum to the calcium channel. PMID- 2554121 TI - Voltage dependence of transient and steady-state Na/K pump currents in myocytes. AB - Experiments are reviewed here in which Na/K pump current was determined as strophanthidin-sensitive current in guinea-pig ventricular myocytes, voltage clamped and internally-dialyzed via wide-tipped pipettes. In the presence of 150 mM extracellular [Na], both outward and inward pump current, during forward and reverse Na/K exchange respectively, were strongly voltage dependent. But reduction of external [Na] to 1.5 mM severely attenuated the voltage sensitivity of outward Na/K pump current. Voltage jumps elicited large transient pump currents during forward or reverse Na/K exchange, or when pump activity was restricted to Na translocation steps, but not when pumps were presumably engaged in K/K exchange. These findings indicate that Na translocation, but not K translocation, involves net charge movement through the membrane field, and that both forward and reverse Na/K transport cycles are rate-limited not by that voltage-sensitive step but by a subsequent voltage-insensitive step. PMID- 2554122 TI - Sodium-hydrogen exchange and its role in controlling contractility during acidosis in cardiac muscle. AB - Intracellular pH (pHi) and Na (aina) were recorded in isolated sheep cardiac Purkinje fibres using ion-selective microelectrodes while simultaneously recording twitch tension. A fall of pHi stimulated acid-extrusion via sarcolemmal Na-H exchange but the extrusion was inhibited by reducing extracellular pH (pHo), indicating an inhibitory effect of external H ions upon the exchanger. Intracellular acidosis can reduce contraction by directly reducing myofibrillar Ca2+ sensitivity. The activation of Na-H exchange at low pHi can offset this direct inhibitory effect of H+ ions since exchange-activation elevates aina which then indirectly elevates Ca2+i (via Na-Ca exchange) thus tending to restore tension. This protection of contraction during intracellular acidosis can be removed if extracellular pH is also allowed to fall since, under these conditions, Na-H exchange is inhibited. PMID- 2554123 TI - Protein phosphorylation and compartments of cyclic AMP in the control of cardiac contraction. AB - The roles of cyclic AMP, cyclic AMP-dependent protein kinase and the phosphorylation of specific proteins in the regulation of cardiac contractility are briefly reviewed. Criteria for determining whether changes in cyclic AMP and protein phosphorylation are involved in a physiological response are discussed. Although cyclic AMP-dependent phosphorylation of the voltage-operated Ca2+ channel, phospholamban, troponin-I and C-protein have all been implicated in the response of the heart to inotropic agents which elevate cyclic AMP, none of these phosphorylations satisfy all of the criteria completely. Evidence is presented that there are compartments of cyclic AMP in heart which are coupled to different functional responses. PMID- 2554124 TI - Transient changes in cyclic AMP and in the enzymic activity of protein kinase and phosphorylase during the cardiac cycle in the canine myocardium and the effect of propranolol. AB - Oscillation of cyclic AMP and in the activity ratio of cyclic AMP-dependent protein kinase and of glycogen phosphorylase with the cardiac cycle were demonstrated in the canine heart in situ. For tissue sampling an ECG (R-wave) triggered, automatically working push-freeze-drill apparatus was developed which allows intraventricular cryobiopsies from the left ventricular muscle of anaesthetized open-chest dogs. The nucleotide cyclic AMP oscillated with the cardiac cycle during normal working condition, the higher cyclic AMP level occuring during systole. Cyclic GMP was assayed to be without oscillatory changes during the contraction-relaxation cycle. The rise in the activity ratio of protein kinase was found to coincide with the maximum in the level of cyclic AMP. Propranolol pretreatment prevents the transient in the level of the nucleotide as well as in the activity ratio of the kinase indicating i) a causal relationship between these changes and ii) a neurohumoral, beat-to-beat regulation by catecholamines released from the sympathetic nerve endings within the heart. Contrary the activity ratio of phosphorylase retains its transient changes during the cardiac cycle in the presence of propranolol, indicating a Ca-mediated activation of phosphorlase kinase during the contraction process. PMID- 2554125 TI - Second messengers involved in the muscarinic control of the heart: the role of the phosphoinositide response. AB - Two heart muscarinic responses are compared, the reduction of cyclic AMP concentration and the hydrolysis of phosphoinositides. It is suggested that the former is more important physiologically, since the latter is a slow response requiring much higher agonist concentrations. Inositol trisphosphate released from phosphoinositides is unlikely to produce a positive inotropic effect by releasing calcium from the sarcoplasmic reticulum. However, long term muscarinic effects may involve activation of protein kinase C by diacylglycerol released from phosphoinositides. PMID- 2554126 TI - Sodium-calcium exchange in mammalian heart: current-voltage relation and intracellular calcium concentration. AB - Membrane currents and changes in intracellular calcium ion concentration ([Ca2+]i) have been recorded that can be attributed to the operation of an electrogenic, voltage-dependent sodium-calcium (Na-Ca) exchanger in mammalian heart cells. Single guinea-pig ventricular myocytes under voltage clamp were perfused internally with the fluorescent Ca2+-indicator, fura-2, and changes in [Ca2+]i and membrane current that resulted from Na-Ca exchange were isolated through the use of various organic channel blockers (verapamil, TTX), impermeant ions (Cs+, Ni2+), and inhibitors of sarcoplasmic reticulum (ryanodine). The I-V relation of Na-Ca exchange was obtained from the Ni2+-sensitive current elicited by ramp repolarization from +90 mV to -80 mV. Ramps were sufficiently rapid that little change in [Ca2+]i occurred during the ramp. The (constant) [Ca2+]i during the ramp was varied over the range 100 nM to 1000 nM by varying the amplitude and duration of a pre-pulse to the ramp. The reversal potential of the Ni2+-sensitive ramp current varied linearly with 1n[( Ca2+])i. The I-V relations at different [Ca2+]i over the range -60 mV to +140 mV were in reasonable accord with the predictions of a simple, simultaneous scheme of Na-Ca exchange, on the basis that only [Ca2+]i had changed. The relationship between [Ca2+]i and current at a constant membrane voltage was also in accord with this scheme. We suggest that Ca2+-fluxes through the exchanger during the cardiac action potential can be understood quantitatively by considering the binding of Ca2+ to the exchanger during the [Ca2+]i-transient and the effects of membrane voltage on the exchanger. PMID- 2554128 TI - [Complete remission of type I diabetes mellitus in infancy following cytomegalovirus infection]. AB - A case of total remission of insulin-dependent Diabetes mellitus in an infant after one year of the disease is described. At time of onset an infection with cytomegalovirus was detected. It is suggested that the protective HLA genes DR2, DQw1 of the patient are responsible for the very rarely seen course of the disease. PMID- 2554127 TI - [Molecular genetic detection of polyclonal immune response in autoimmune thyroiditis and inflammatory bowel diseases]. AB - Southern blot hybridization techniques were used to analyze the arrangements of the immunoglobulin and the T cell antigen receptor genes in lymphocytes of patients with Graves disease and Hashimoto's thyroiditis as well as in patients with Crohn's disease, chronic ulcerative colitis, and with other gastrointestinal disease. The results indicate that the immune response of autoimmune thyroid disease and inflammatory bowel disease is of polyclonal origin. PMID- 2554129 TI - Neuronal Na+, K+-ATPase in the peripheral nerve fibers. AB - ATPase activity was localized by means of Wachstein-Meisel's method in rat sciatic nerve fibers. Using controls with ouabain, the presence of alpha + (neuronal) Na+, K+-ATPase was examined. The enzyme occurs in the ATPase reaction of the myelin-forming membranes, axoplasm and Schwann cell cytoplasm. Its presence in the Schwann cell plasma membrane is only admittable. The ATPase activity of the compact myelin and axolemma was exclusively of alpha + type of Na+, K+-ATPase. PMID- 2554130 TI - [New producers of site-specific endonucleases from microorganisms of the Bacillus genus]. AB - 52 strains of Bacillus generum have been tested for production of site-specific endonucleases. The sequence recognized by the enzyme was determined for 23 enzymes, the cleavage site inside the sequence was determined for 5 enzymes. All the enzymes under study were found to be isomers of the known enzymes. The selected strains are peculiar for the high level of site-specific endonucleases content and may be used as producents of the enzymes. PMID- 2554131 TI - [Immunoenzyme analysis of adenovirus hexons. Indication and characteristics of rabbit and mouse antibodies against hexons]. AB - An enzyme-linked immunosorbent assay (ELISA) was developed for analysis of rabbit and mouse IgG antibodies specific to adenoviral hexon. The anti-hexon antibodies were detected by capture with purified hexon coated onto polystyrene microtiter plates and visualizing them by respective anti-IgG horseradish peroxidase conjugates. In the sera from hyperimmunized rabbits and mice as well as in the mouse ascite fluids the ELISA procedure revealed primarily type-specific (epsilon) and genus-specific (alpha) antigenic determinants in hexon but not those of intermediate specificities. PMID- 2554132 TI - [Vaccinia and ectromelia recombinant viruses, causing an infection, characteristic for ectromelia, in mice]. AB - Ten recombinants between the viruses of vaccinia and ectromelia were isolated that cause the ectromelia virus specific lesions in mice. The structure of recombinant viral genomes, the efficiency of viral propagation in mice, the nature of lesions induced by viruses have been studied. Eight of obtained recombinants have a DNA insertion originating from the right end of ectromelia viral genome, nine recombinants have an insertion originating from the left end, seven recombinants possess both insertions. The latter recombinants have more pronounced pathogenicity for mice. Both revealed regions are supposed to define the specific pathogenicity of ectromelia virus for mice. PMID- 2554133 TI - The interaction between X-rays and transposon mobility in Drosophila: hybrid sterility and chromosome loss. AB - Genetic traits associated with P-M hybrid dysgenesis in Drosophila melanogaster were synergistically affected by X-rays. The interaction between damages induced by these two mutator systems was evident when sterility and X/Y chromosome loss were used as endpoints. No interaction was detected in partial chromosome loss, monitored by the loss of BS and y+ markers. The synergism in sterility, measured either as all-or-none or premature sterility (fecundity) was observed when male hybrids derived from different P strains fathers, namely Harwich or II2, were X irradiated and the effects compared relative to similarly treated non-dysgenic hybrids. Brooding of sperm showed that the effects of ionizing radiation were ionizing radiation were dependent upon the stage of spermatogenesis during which cells were irradiated. The highly synergistic effect on sterility was found when either spermatids or spermatocytes, but not mature sperm, were irradiated with 550 rad of X-rays. These findings were consistent with the higher radiosensitivity of spermatocytes and spermatids to genetic damage and with the correlation between the incidence of sterility and aging of dysgenic hybrids. The latter observation was particularly evident in the case of Harwich P strain derived male hybrids whose fertility was greatly reduced due to P element mobility. The synergistic effect of X-rays in these dysgenic hybrids resulted in the virtual abolition of the germ line, increasing the sterility from 50% of the untreated 9-10-day old males, to 85% of the treated males when spermatocytes were irradiated. The synergism observed between transposon mobility and ionizing radiation can best the attributed to an interaction between X-ray-induced and P element-induced chromosome breaks. This interpretation is consistent with the more than additive increase in X or Y chromosome loss in irradiated, Harwich P strain-derived hybrid sons. The induction of these events was 1.164% in dysgenic irradiated males as compared to 0.234% in X-irradiated nondysgenic hybrids and 0.40% in dysgenic untreated males. No synergism was observed in X/Y loss in hybrids derived from II2 P strain fathers where the frequency of the events due to P element mobility alone was only one tenth (0.037%) of that found in Harwich derived hybrids. PMID- 2554135 TI - DNA topoisomerases and models of sister-chromatid exchange. AB - Pommier et al. (1985) suggested that sister-chromatid exchange (SCE) results from exchange of topoisomerase II subunits. "Homologous displacement", an alternative mechanism, is proposed in which strand switching occurs during removal of parental helical turns by topoisomerases. The steps in the SCE model proposed by Ishii and Bender (1980) for SCE occurring at a blocked replication fork could occur by this mechanism and would require the action of both topoisomerases I and II. Homologous displacement involving topoisomerase II alone provides a mechanism for the strand switching required in the models of Kato (1977) and Cleaver (1981) in which SCE occur between replicated double strands. These mechanisms and models are discussed in relation to current knowledge of the locations and functions of topoisomerases during DNA replication. PMID- 2554134 TI - Role of ruvAB genes in UV- and gamma-radiation and chemical mutagenesis in Escherichia coli. AB - Escherichia coli umuC122::Tn 5 was mutagenized with N-methyl-N'-nitro-N nitrosoguanidine to isolate mutations that block the residual gamma-radiation mutagenesis observed in umuC strains. Two of these mutations were shown by transductional mapping and plasmid complementation to map in the ruvA and ruvB genes (i.e., ruvA200 and ruvB201). Whereas ruvA200 was complemented by ruvA+ plasmids, the only other known ruvA mutation, ruvA59::Tn10 required both the ruvA+ and ruvB+ genes to show complementation. The ruvA200, ruvB201, ruvA59::Tn10 and ruvB60::Tn10 mutations all reduced gamma-radiation-induced ochre reversion [argE3(Oc)----Arg+] to about 30% of the wild-type level, and they all reduced UV radiation-induced ochre reversion to about 15% of the wild-type level. The ruvA200 and ruvB201 mutants also showed reduced gamma- and UV-radiation mutagenesis with two other assays [hisG4(Oc)----His+ and Rifs----Rifr]. Streptozotocin mutagenesis (Rifr) was reduced to about half of the wild-type level in ruv strains, but ethyl methanesulfonate mutagenesis was normal. While the umuC strain did not show the oxygen enhancement of gamma-radiation mutagenesis, the ruvA200 strain showed an oxygen effect that was similar to that shown by the wild-type strain. When the ruvA200 mutation was combined with the umuC mutation, gamma-radiation mutagenesis was further reduced to 5% of the wild type level and cells showed a synergistic sensitization to UV- and gamma radiation-induced killing. A mutational spectrum analysis indicates a general depression of both umuC-dependent and umuC-independent gamma-radiation mutagenesis in the ruvA strain, which is in contrast with the site-specific reduction in gamma-radiation mutagenesis that is observed in the umuC mutant. The reduced radiation mutagenesis in the ruvA strain could not be correlated with a reduction in transcription of the recA or umuC genes. PMID- 2554136 TI - Comparison of the nucleotide sequences of a yeast gene family. II. Analysis of spontaneous deletions and insertions. AB - We compared the nucleotide sequences of 3 yeast invertase genes in regions where the homology is better than 90%. In the noncoding region 40 gaps of 1-61 bases were found. This is about half as much as the nucleotide substitutions in the same sequences. We grouped the gaps into 5 categories by their length and the characteristics of their sequences. Group I gaps are about 20 nucleotides long and are flanked by repeated sequence of 6 bases which may trigger the deletion of one of the repeats and the sequence between the repeats. Group II gaps are characterized by a small repeated sequence which is missing in one of the invertase genes. Gaps which occur in sequences exclusively made up of one of the 4 bases are summarized in group III. The 4 gaps in group IV do not show any of these sequence characteristics and they are all just one base long. A 61 nucleotide sequence found in only one of the invertase genes seems to be of complex origin. We conclude that small repeated sequences or monotonous sequences are prone to deletion or insertion mutations. PMID- 2554137 TI - Beta-galactosidase overexpression in SV40-transformed Chinese hamster fibroblasts exposed to mutagens as a result of amplification of transfected bacterial lacZ DNA sequences. AB - Genetic constructions in which the bacterial lacZ gene, encoding the enzyme beta galactosidase, is fused to a viral (SV40) origin of replication have been introduced in an SV40-transformed hamster cell line (C1102). We have studied in detail 3 clones in which beta-galactosidase-specific activity increases after treatment with genotoxic agents. We show that this increase is dependent on the activity of the viral T protein and correlates with an amplification of lac sequences. This system provides a basis for the study of the induction of gene amplification by genotoxic agents in mammalian cells. PMID- 2554138 TI - Thymidine kinase deficient human cells have increased UV sensitivity in their capacity to support herpes simplex virus but normal UV sensitivity for colony formation. AB - A thymidine kinase deficient (tk-) and two thymidine kinase proficient (tk+) human cell lines were compared for UV sensitivity using colony-forming ability as well as their capacity to support the plaque formation of herpes simplex type 1 (HSV-1). The tk- line (143 cells) was a derivative of one of the tk+ lines (R970 5), whereas the other tk+ line (AC4 cells) was a derivative of the 143 cells obtained by transfection with purified sheared HSV-2 DNA encoding the viral tk gene. 143, R970-5 and AC4 cells showed a similar UV sensitivity for colony forming ability. In contrast, the capacity to support HSV-1 plaque formation immediately (within 1 h) after UV-irradiation was reduced to a greater extent in the 143 cells compared to the R970-5 and AC4 cells. Capacity curves for plaque formation of the HSV-1: KOS wild-type (tk+) strain were similar to those for the HSV-1: PTK3B mutant (tk-) strain in the 3 cell strains, indicating that the viral tk gene does not influence the ability of HSV-1 to form plaques in UV-irradiated compared to unirradiated human cells. Cellular capacity for HSV-1 plaque formation was found to recover in both tk- and tk+ cells for cultures infected 24 h after UV-irradiation. These results suggest that repair of UV-damaged DNA takes place to a similar extent in both tk- and tk+ human cells, but the kinetics of repair are initially slower in tk- compared to tk+ human cells. PMID- 2554140 TI - Early employment testing for marijuana: demographic and employee retention patterns. PMID- 2554139 TI - UV-enhanced reactivation of UV-damaged SV40 is due to the restoration of viral early gene function. AB - Mammalian cells respond to UV-radiation by inducing an increased ability to support the survival of UV-damaged virus. We have tested whether the induction of enhanced viral reactivation (ER) reflects heightened UV-resistance of specific viral functions. For this, we examined the extent of ER for SV40 containing UV damage in three functionally distinct regions of the SV40 genome: (i) the viral regulatory region, (ii) the early genes region and (iii) the late genes region. ER corresponding to a dose reduction factor of 43% was observed for damage in the early genes region. No ER was observed for damage in the regulatory or late genes regions. We conclude that ER in SV40 reverses the lethal disruption of an essential function peculiar to the viral early genes region. This function is almost certainly transcription. PMID- 2554141 TI - Calcium signalling: receptor kinships revealed. PMID- 2554142 TI - Primary structure and functional expression of the inositol 1,4,5-trisphosphate binding protein P400. AB - Cloning and expression of functional P400 protein from cerebellar Purkinje neurons shows that this protein is a receptor for inositol 1,4,5-trisphosphate, a second messenger that mediates the release of intracellular calcium. PMID- 2554143 TI - Purified inositol 1,4,5-trisphosphate receptor mediates calcium flux in reconstituted lipid vesicles. AB - Inositol 1,4,5-trisphosphate (Ins(1,4,5)P3), a second messenger molecule involved in actions of neurotransmitters, hormones and growth factors, releases calcium from vesicular non-mitochondrial intracellular stores. An Ins(1,4,5)P3 binding protein, purified from brain membranes, has been shown to be phosphorylated by cyclic-AMP-dependent protein kinase and localized by immunohistochemical techniques to intracellular particles associated with the endoplasmic reticulum. Although the specificity of the Ins(1,4,5)P3 binding protein for inositol phosphates and the high affinity of the protein for Ins(1,4,5)P3 indicate that it is a physiological Ins(1,4,5)P3 receptor mediating calcium release, direct evidence for this has been difficult to obtain. Also, it is unclear whether a single protein mediates both the recognition of Ins(1,4,5)P3 and calcium transport or whether these two functions involve two or more distinct proteins. In the present study we report reconstitution of the purified Ins(1,4,5)P3 binding protein into lipid vesicles. We show that Ins(1,4,5)P3 and other inositol phosphates stimulate calcium flux in the reconstituted vesicles with potencies and specificities that match the calcium releasing actions of Ins(1,4,5)P3. These results indicate that the purified Ins(1,4,5)P3 binding protein is a physiological receptor responsible for calcium release. PMID- 2554144 TI - Yeast replication factor-A functions in the unwinding of the SV40 origin of DNA replication. AB - Cell-free replication systems for simian virus 40 (SV40) DNA are taken to be a model for the replication of eukaryotic chromosomes, because only one viral protein is required to supplement the replication proteins provided by a human cell extract. To prove that these cellular proteins function in chromosomal DNA replication we have begun to identify homologous proteins in an organism that can be genetically manipulated. Here we report the identification of yeast replication factor-A (yRF-A) from Saccharomyces cerevisiae and show that it is functionally and structurally related to a human protein that is required for the initiation and elongation of SV40 DNA replication. Yeast RF-A, a multi-subunit phosphoprotein, is similar to the human protein in its chromatographic behaviour, subunit structure and DNA-binding activity. The yeast protein will fully substitute for the human protein in an early stage of the initiation of SV40 DNA replication. Substitution of yRF-A in the complete SV40 replication system, however, results in reduced DNA replication, presumably due to a requirement for species-specific interactions between yeast RF-A and the DNA polymerase complex. PMID- 2554145 TI - Induction of the Escherichia coli lactose operon selectively increases repair of its transcribed DNA strand. AB - Nucleotide excision repair helps to ameliorate the lethal and mutagenic consequences of DNA damage by removing helix-distorting lesions from cellular genomes. We have previously analysed the removal of ultraviolet-induced cyclobutane pyrimidine dimers from specific DNA sequences in mammalian cells and demonstrated that transcriptionally active genes are preferentially repaired. Additionally, we found that in rodent and human cells only the transcribed strand of the dihydrofolate reductase gene is selectively repaired. Transcription is blocked by pyrimidine dimers in template DNA and the selective removal of these lesions seems to be important for cell survival after irradiation with ultraviolet light. To determine whether this feature of repair is common to prokaryotes and eukaryotes and better to understand its mechanism, we have investigated repair in the two separate DNA strands of the lactose operon of ultraviolet-irradiated Escherichia coli. We find a dramatic difference in the repair of the two strands only when transcription is induced. Most dimers are removed from the transcribed strand of the induced operon within five minutes of irradiation. In the nontranscribed strand, repair is significantly slower and resembles that found in both strands of the uninduced operon. Thus there seems to be a mechanism that couples nucleotide excision repair and transcription. PMID- 2554146 TI - Putative receptor for inositol 1,4,5-trisphosphate similar to ryanodine receptor. AB - Inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) serves as an intracellular second messenger for several neurotransmitters, hormones and growth factors by initiating calcium release from intracellular stores. A cerebellar Ins(1,4,5)P3 receptor has been characterized biochemically and shown by immunocytochemistry to be present in intracellular membranes in Purkinje cells. We show that a previously described Purkinje-cell messenger RNA encodes a protein of relative molecular mass 260,000 (260 K) with the same properties as the cerebellar Ins(1,4,5)P3 receptor. Its sequence is partially homologous to the skeletal muscle ryanodine receptor. By immunocytochemistry and electron microscopy the protein is shown to be present in all parts of the endoplasmic reticulum, including those that extend into axon terminals and dendritic spines. Our results indicate that gated calcium release from intracellular stores in muscle and Purkinje cells uses similar calcium-channel proteins localized in analogous intracellular compartments. This implies that the intracellular calcium stores in the endoplasmic reticulum of neurons extend into presynaptic terminals and dendritic spines where they may play a direct role in regulating the efficacy of neurotransmission. PMID- 2554147 TI - FMRFamide reverses protein phosphorylation produced by 5-HT and cAMP in Aplysia sensory neurons. AB - Neurotransmitter can modulate neuronal activity through a variety of second messengers that act on ion channels and other substrate proteins. The most commonly described effector mechanism for second messengers in neurons depends on protein phosphorylation mediated by one of three sets of kinases: the cyclic AMP dependent protein kinases, the Ca2+-calmodulin-dependent protein kinases, and the Ca2+-phospholipid-dependent protein kinases. In addition, some neurotransmitters and second messengers can also inhibit protein phosphorylation by lowering cAMP levels (either by inhibiting adenylyl cyclase or activating phosphodiesterases). This raises the question: can neurotransmitters also modulate neuronal activity by decreasing protein phosphorylation that is independent of cAMP? Various biochemical experiments show that a decrease in protein phosphorylation can arise through activation of a phosphatase or inhibition of kinases. In none of these cases, however, is the physiological role for the decrease in protein phosphorylation known. Here we report that in Aplysia sensory neurons, the presynaptic inhibitory transmitter FMRFamide decreases the resting levels of protein phosphorylation without altering the level of cAMP. Furthermore, FMRFamide overrides the cAMP-mediated enhancement of transmitter release produced by 5-hydroxytryptamine (5-HT), and concomitantly reverses the cAMP-dependent increase in protein phosphorylation produced by 5-HT. These findings indicate that a receptor-mediated decrease in protein phosphorylation may play an important part in the modulation of neurotransmitter release. PMID- 2554148 TI - Requirement of the Drosophila raf homologue for torso function. AB - In Drosophila the correct formation of the most anterior and posterior regions of the larva, acron and telson is dependent on the maternally expressed terminal class of genes. In their absence, the anterior head skeleton is truncated and all the structures posterior to the abdominal segment seven are not formed. The protein predicted to be encoded by one of these genes, torso (tor), seems to be a transmembrane protein with an extracytoplasmic domain acting as a receptor and a cytoplasmic domain containing tyrosine kinase activity. Here we report that another member of the terminal-genes class, l(1)polehole (l(1)ph), which is also zygotically expressed, is the Drosophila homologue of the v-raf oncogene and encodes a potential serine-and-threonine kinase. We also show that functional l(1)ph gene product is required for the expression of a gain-of-function tor mutant phenotype, indicating that l(1)ph acts downstream of tor. Together, these results support the idea that the induction of terminal development occurs through a signal transduction system, involving the local activation of the tor encoded tyrosine kinase at the anterior and posterior egg poles, resulting in the phosphorylation of the l(1)ph gene product. In turn, downstream target proteins may be phosphorylated, ultimately leading to the regionalized expression of zygotic target genes. Such a process is in agreement with the finding that both tor and l(1)ph messenger RNAs are evenly distributed. PMID- 2554149 TI - Adrenaline-induced enhancement of the blood pressure response to sympathetic nerve stimulation in adrenal demedullated pithed rats. AB - The influence of the prejunctional beta-adrenoceptor-mediated control mechanism of noradrenaline release on the cardiovascular response to preganglionic nerve stimulation (PNS) and the role of adrenaline as endogenous activator of this facilitatory mechanism were studied in pithed rats. The increases in mean arterial blood pressure and heart rate responses to eight PNS (0.8 Hz, 1 ms, 75 V, for 20 s at 12 min intervals) were measured in control, adrenal demedullated and sham-operated pithed rats, respectively. Repeated stimuli of the sympathetic outflow elicited similar blood pressure responses in control and sham-operated rats, whereas the blood pressure responses were attenuated in adrenal demedullated rats. Administration of the non-selective beta-adrenoceptor antagonist propranolol (5 mg x kg-1, i.v.) decreased the blood pressure response in control, but not in adrenal demedullated rats. The increase in heart rate during PNS was abolished in both groups after administration of propranolol. Infusion of adrenaline (0.06 microgram x kg-1 x min-1) increased the circulation levels of adrenaline from 0.4 nmol x l-1 to 4.8 nmol x l-1, but did not modify the blood pressure response to PNS in control rats. In adrenal demedullated rats, however, the blood pressure response to PNS was enhanced during as well as after the infusion of adrenaline. This facilitatory effect of adrenaline on the blood pressure response in adrenal demedullated rats was blocked by pretreatment with propranolol (5 mg x kg-1, i.v.). Taken together the present findings give further support to the hypothesis that neuronal adrenaline of adrenal medullary origin is the endogenous agonist at prejunctional beta-adrenoceptors mediating a facilitation of neuronal noradrenaline release. PMID- 2554151 TI - Effects of 1,3-dipropyl-8-cyclopentylxanthine (DPCPX), a highly selective adenosine receptor antagonist, on force of contraction in guinea-pig atrial and ventricular cardiac preparations. AB - The effects of the A1 adenosine receptor antagonist 1,3-dipropyl-8 cyclopentylxanthine (DPCPX) on force of contraction were examined in isolated electrically driven auricles and papillary muscles from guinea-pigs in the absence and presence of (-)-N6-phenylisopropyladenosine (PIA) and 5'-N ethylcarboxamidadenosine (NECA). In auricles DPCPX (30-1000 mmol/l) alone increased force of contraction. DPCPX produced only a minor inhibition of phosphodiesterase I-III activity. PIA and NECA alone exerted concentration dependent negative inotropic effects and the concentration-response curves for PIA and NECA were shifted competitively to the right by the adenosine receptor antagonist DPCPX with similar potency and efficacy. The pA2-value for the inhibition of the effects of PIA and NECA were 9.1 and 8.8, respectively. In papillary muscles DPCPX alone had no inotropic effect. In the presence of isoprenaline PIA and NECA alone exerted concentration-dependent negative inotropic effects and again DPCPX shifted the concentration-response curves for PIA and NECA competitively to the right with similar potency and efficacy. The pA2-value for the inhibition of the effects of PIA and NECA were 9.3 and 9.0, respectively. It is concluded that DPCPX is a potent competitive A1 adenosine receptor antagonist in guinea-pig atrial and ventricular cardiac preparations. Since PIA and NECA were equally potent the cardiac adenosine receptor may constitute a subtype of A1 adenosine receptors differing from the receptor in other tissues such as fat cells. Furthermore, DPCPX has a positive inotropic effect in atrial tissue which cannot be attributed to the A1 receptor antagonism. PMID- 2554150 TI - Dissociation of phosphoinositide hydrolysis and positive inotropic effect of histamine mediated by H1-receptors in guinea-pig left atria. AB - The present study was undertaken to determine whether the phosphoinositide hydrolysis is responsible for the positive inotropic effect of histamine in guinea-pig left atria. Histamine induced hydrolysis of phosphoinositides and a positive inotropic effect in a concentration-dependent manner. These effects were antagonized by chlorpheniramine (0.1 mumol/l) but not by cimetidine (10 mumol/l). At a concentration of 1 mumol/l histamine produced a dual-component positive inotropic response composed of an initial increasing phase and a second and late developing, greater positive inotropic phase. Histamine (10 mumol/l) caused a gradual increase in the formation of [3H]inositol trisphosphate (IP3) and a significant increase in the [3H]IP3 level was detected 10 min after the stimulation. Thus, the increase in IP3 did not precede the increase in force of contraction. The phospholipase C inhibitors 2-nitro-4-carboxyphenyl-N,N diphenylcarbamate (100 mumol/l) and neomycin (100 mumol/l) significantly reduced the histamine-induced [3H]inositol monophosphate accumulation. However, pretreatment with the phospholipase C inhibitors did not affect the positive inotropic effect of histamine, either in its extent or in its pattern. The phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) (100 nmol/l) and phorbol 12,13-dibutyrate (PDBu) (100 nmol/l) also significantly inhibited the phosphoinositide hydrolysis induced by histamine. The inhibitory effect of the phorbol esters on the phosphoinositide response was completely abolished in the presence of 10 mumol/l 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), a protein kinase C inhibitor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554152 TI - Cardiac Ca current does not run down and is very sensitive to isoprenaline in the nystatin-method of whole cell recording. AB - The conventional L type Ca2+ channel current (ICa.L) was measured in single atrial and ventricular myocytes, with a new whole-cell recording technique using an ionophore, nystatin. The membrane of a cell-attached patch was gradually permeabilized by nystatin (100-200 micrograms/ml), added to the pipette solution, within 2-5 min after formation of a G omega-seal. The electrical activity of the cells was measured through the pipette. ICa.L, measured with the nystatin-whole cell recording technique, did not exhibit the so-called "run-down" phenomenon for up to 90 min. The response of ICa.L to isoprenaline was also well preserved during the measurement. The half maximal concentration for the isoprenaline induced increase of ICa.L was 8.2 x 10(-9) M, which is a much smaller value than that reported previously. Thus, the nystatin-whole cell clamp recording is a useful technique to measure membrane currents of cardiac myocytes with preserving the physiological intracellular milieu. PMID- 2554153 TI - Modulation of noradrenaline release by presynaptic alpha-2 and beta adrenoceptors in rat atria. Effect of pretreatment with clenbuterol. AB - The effect of pretreatment with the beta-2-selective adrenoceptor agonist, (+/-) clenbuterol (0.3 mg/kg, twice daily, 14 days) on prejunctional alpha-2- and beta adrenoceptors was studied in rat atria. When atria from non-pretreated rats had been preincubated with (3H)-noradrenaline, (-)-isoprenaline (0.02 to 4.0 microM) did not affect tritium overflow evoked by stimulation of the cardioaccelerant nerves, but a higher concentration (40 microM) decreased it. Blockade of prejunctional inhibitory alpha-2-adrenoceptors by yohimbine (0.03, 0.3 and 0.8 microM) enhanced the overflow of tritium. In the presence of yohimbine, isoprenaline (1.2 microM) significantly increased stimulation-induced transmitter overflow, suggesting that in rat atria the facilitatory effect of isoprenaline mediated via prejunctional beta-adrenoceptors, is masked by the dominant influence of inhibitory alpha-2-adrenoceptors. (-)-Propranolol (0.1 microM) prevented the isoprenaline-induced increase in atrial rate and the isoprenaline induced enhancement of transmitter release in the presence of yohimbine (0.3 microM), but did not modify by itself the stimulation-induced efflux of tritium, suggesting that neuronally released noradrenaline failed to activate facilitatory prejunctional beta-adrenoceptors. When atria from clenbuterol-pretreated rats had been preincubated with 3H-noradrenaline, the facilitatory effect of yohimbine 0.03 and 0.3 microM was markedly enhanced and, in this case, isoprenaline (1.2 and 12.0 microM) failed to cause its facilitatory effect in the presence of the alpha-2-adrenoceptor antagonist. Propranolol did not modify the facilitatory effect of yohimbine. No changes in the isoprenaline-induced increase in atrial rate were observed in clenbuterol-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554154 TI - The negative inotropic effect of neuropeptide Y on the ventricular cardiomyocyte. AB - The effect of neuropeptide Y (NPY) on cell contractions of ventricular myocytes isolated from the adult rat heart was investigated. Maximum changes in cell length (dL) during stimulated (0.5 Hz) contractions were determined in presence of the phosphodiesterase inhibitor Ro 20-1724 (0.5 mM) and adenosine deaminase (5 U/ml). Under these basal conditions NPY (10(-6) M) reduced dL by 39% of control. Isoproterenol (10(-6) M) increased dL by 105% of control; the EC50 was 2 x 10(-9) M. NPY reduced the increase in dL achieved by isoproterenol in a dose dependent manner. The IC50 value was 1 x 10(-9) M and NPY (10(-6) M) produced complete inhibition. In the absence of the phosphodiesterase inhibitor the IC50 was 4 x 10(-9) M. The EC50 of isoproterenol and IC50 of NPY producing accumulation of cAMP in myocytes (Millar et al. 1988) exceeded the respective values of dL by one order of magnitude. Prior treatment of the myocytes with pertussis toxin abolished the potency of NPY to antagonize the increase in dL by isoproterenol while not interfering with the response to the beta-agonist. These results demonstrate a negative inotropic effect of NPY on the ventricular myocardial cell. Complete abolition of the effect of NPY by pertussis toxin indicate that this effect is mediated by a sarcolemmal receptor for NPY linked to adenylate cyclase via an inhibitory guanine nucleotide binding protein. PMID- 2554156 TI - [Is there a role for radiotherapy in the treatment of patients with pleomorphic adenoma of the parotid gland?]. AB - A group of 79 patients primarily operated (n = 63) or operated for local recurrence after previous surgery (n = 16) and irradiated post-operatively was studied retrospectively. Indications for radiation treatment were: enucleation and spill after pseudo-penetration of the capsule or remnants of tumour after partial or total parotidectomy respectively. During follow-up (4-16 years) only one of the patients, irradiated because of recurrence, had a second recurrence. No major complications or malignant degenerations took place. Partial or total parotidectomy with saving of the facial nerve is the treatment of choice rather than a combination of enucleation with radiotherapy. However, in case of remnants after radical surgery or re-excision for recurrent tumour postoperative irradiation appears to be indicated and effective. PMID- 2554157 TI - [Prolonging of life with cytostatic treatment: is it worthwhile?]. PMID- 2554155 TI - Ventricular and atrial electrophysiological effects of a IC antiarrhythmic drug, cibenzoline, in the innervated dog heart. Role of sodium and calcium channels. AB - The effects of cibenzoline, rightly known as a sodium channel inhibitor (class IC antiarrhythmic drug), were investigated in anaesthetized, closed-chest dogs, on conduction in the contractile fibres, ventricular and atrial, the His-Purkinje system and the atrioventricular node. In ventricular muscle, conduction time was measured between base and apex by two endocavitary electrodes. The other conduction times were obtained from the recording of the His bundle potentials. In addition, effective refractory period was determined by the extrastimulus method in ventricular and atrial muscle and in the atrioventricular node, and sinus rate monitored in the intervals of pacing periods. In the absence of vagal tone, cibenzoline in 4 mg.kg-1 dose prolonged conduction times in the ventricular contractile tissue, His-Purkinje system and atrial contractile tissue to a large extent, but decreasingly from the former to the latter. This prolongation was antagonized by hypernatremia (174 mmol.l-1). In contrast, conduction time in the atrioventricular node, effective refractory periods and sinus rate were very little influenced. In the presence of vagal tone, the prolongation of conduction times in the ventricular contractile tissue. His-Purkinje system and atrial contractile tissue did not differ substantially from previously. It was the same for ventricular effective refractory period. But atrial effective refractory period was then considerably lengthened, while conduction time and effective refractory period in the atrioventricular node were greatly shortened and sinus rate notably accelerated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554159 TI - Alternative approach. Interview by Tim Rice. PMID- 2554158 TI - [The interval carcinoma in periodic breast screening, a new concept]. PMID- 2554160 TI - [Subsynaptic modulation of chemical stimulation transmission by metacerebral neurons in the buccal ganglia of mollusks]. AB - Stimulation of giant metacerebral serotonin-containing cell LMc1 in Helix pomatia was studied for its effect on responses of buccal cells LBc2 and LBc3 to stimulation of buccal nerves. Stimulation of cell LMc1 modulated those responses either increasing or decreasing them. Results of applications of serotonin, dibutyryl-cAMP, methysergide, d-tubocurarine and acetylcholine on cells LBc2 and LBc3 suggest that the modulation of responses of buccal cells LBc2 and LBc3 to buccal nerves stimulation by cell LMc1 was subsynaptic. PMID- 2554161 TI - [Potentiating and blocking action of cyclic adenosine monophosphate and lithium ions on mollusk neuronal responses caused by acetylcholine and gamma-aminobutyric acid]. AB - It was found that cyclic adenosine monophosphate and lithium ions produced blocking effects on acetylcholine-induced currents carried by sodium, potassium and calcium ions in neurons of Helix pomatia. Lithium ions produced biphasic actions on ACh-induced and gamma-aminobutyric acid-induced chloride currents. PMID- 2554162 TI - Selective decontamination of the digestive tract in hematological patients (Czechoslovakia-German Democratic Republic cooperative study. AB - A group of 55 hematological patients treated for the last 2.5 years by the method of selective decontamination was evaluated. Though both institutes (Bad Saarow, Hradec Kralove) worked on the problem in the same conditions (indications for the treatment, characteristics of patients, basic drugs), many differences in details were found. However, the important clinical results were the same: A statistically significant decrease in infections and duration of fever in treated patients. A survey of therapy complications, surveillance of infections and incidence of microbes are presented. The evaluation showed that future research including microbiological and immunological investigation based on a standard protocol will be useful. PMID- 2554163 TI - [Comments on the contribution by G. F. Walter et al. Detection of Epstein-Barr viruses in Rasmussen encephalitis]. PMID- 2554165 TI - Serum angiotensin-converting enzyme as a marker of dialyzer membrane biocompatibility? PMID- 2554164 TI - Effects of dialysis and transplantation on red cell Na pump function in renal failure. AB - Ion pumping by the erythrocyte Na, K-ATPase has been measured using ouabain sensitive 86Rb flux in 11 non-dialysed patients with chronic renal failure (CRF), 13 patients on haemodialysis (HD), 13 patients on peritoneal dialysis (CAPD) and 15 patients with functional transplants (FT). Flux measurements were performed in plasma and simultaneous estimates of specific 3H-ouabain binding were made. The results indicate that, compared to normal controls, Na,K pump flux was reduced by 21% in CRF (p less than 0.01), 30% in HD (p less than 0.01), 15% in CAPD (p less than 0.02), and was normal in FT. Mean specific ouabain binding sites per cell (+/- SEM) were; controls 366 +/- 16; CRF, 290 +/- 16; HD, 344 +/- 17; CAPD, 321 +/- 18; FT, 345 +/- 26. Calculation of mean turnover rate per pump site indicated that patients on HD showed a 30% reduction compared to controls (influx 55 K ions/s versus 79 K ions/s, p less than 0.01). Cross-incubation experiments suggest that the lowered pump flux seen in the CRF and HD groups was due to plasma factors. This work shows that erythrocyte Na,K pump number is reduced in CRF, while patients on maintenance HD have normal pump numbers per erythrocyte but reduced pump turnover. PMID- 2554166 TI - Age-associated alterations in catecholaminergic concentrations, neuronal activity, and alpha 1 receptor densities in female rats. AB - The density of alpha 1-adrenergic receptors and catecholamine concentrations and neuronal activity were assessed in selected estrogen-responsive regions of the hypothalamus and pineal glands. The results of this study demonstrate that the densities of alpha 1-adrenergic receptors and norepinephrine and epinephrine concentrations and activity rate constants are altered in aged rats. The direction and extent of the change depend on the specific brain region and reproductive state of the animals. No widespread decline with increased age was observed in any of the parameters measured. PMID- 2554167 TI - Selective improvement of aged rat short-term spatial memory by 3,4 diaminopyridine. AB - Young (10 month) and old (28 month) Fischer 344 rats were injected (IP) with 3,4 diaminopyridine (3,4-DAP) or saline 10 minutes before training on two tests of spatial memory (the Barnes circular platform and the radial 8-arm maze). This agent has been found to block potassium channels in neurons, thereby increasing calcium influx, prolonging the action potential, and leading to increased transmitter release. The circular platform task assessed the drug's effect on spatial reference memory over 24 hour intertrial intervals, and the radial maze assessed its effect on short-term working memory within a 5 minute test session. 3,4-DAP was found to selectively improve memory performance of the old animals, and, within that age group, only improved performance on the short-term memory task. 3,4-DAP may therefore be effective for only a restricted set of age-related memory problems. PMID- 2554169 TI - Non-NMDA receptor-mediated neuronal injury in Alzheimer's disease? AB - The foregoing review by J. T. Greenamyre and A. B. Young provides an excellent discussion of the alterations in excitatory amino acid transmitter systems that accompany Alzheimer's disease, and the interesting possibility that excitotoxicity may participate in disease pathogenesis. I would add a speculation that non-NMDA receptors may be particularly important in mediating such excitotoxicity. PMID- 2554168 TI - Excitatory amino acids and Alzheimer's disease. AB - Excitatory amino acids (EAA) such as glutamate and aspartate are major transmitters of the cerebral cortex and hippocampus, and EAA mechanisms appear to play a role in learning and memory. Anatomical and biochemical evidence suggests that there is both pre- and postsynaptic disruption of EAA pathways in Alzheimer's disease. Dysfunction of EAA pathways could play a role in the clinical manifestations of Alzheimer's disease, such as memory loss and signs of cortical disconnection. Furthermore, EAA might be involved in the pathogenesis of Alzheimer's disease, by virtue of their neurotoxic (excitotoxic) properties. Circumstantial evidence raises the possibility that the EAA system may partially determine the distribution of pathology in Alzheimer's disease and may be important in producing the neurofibrillary tangles, RNA reductions and dendritic changes which characterize this devastating disorder. In this article, we will review the evidence suggesting a role for EAA in the clinical manifestations and pathogenesis of Alzheimer's disease. PMID- 2554170 TI - Selective neuronal vulnerability and the distribution of N-methyl-D-aspartate (NMDA) receptors. AB - N-Methyl-D-Aspartate (NMDA) receptors are believed to play a critical role in excitotoxic cell death in the CNS. The distribution of NMDA-preferring binding sites is compared here with the patterns of selective neuronal death observed in Alzheimer's disease and following transient ischemia. The distribution of NMDA receptors, by itself, is unable to account for the characteristic patterns of selective neuronal vulnerability observed in conjunction with these types of neuropathology. PMID- 2554171 TI - Excitatory amino acids and Alzheimer's disease: idle thoughts on an exciting subject. AB - The reported loss of cortical glutaminase activity in Alzheimer's disease is another possible indicator of loss of glutamate neurons. A diversity of excitatory amino acids (EAA) and NMDA receptor subtypes might explain selective neuronal losses of neurons in various diseases. Weaknesses in the arguments presented are the multiple actions of THA and that 2-amino-3 (methylamino)propionate is probably not an EAA. PMID- 2554172 TI - Stimulation of phosphoinositide degradation and phosphatidylinositol-4-phosphate phosphorylation by GTP exclusively in plasma membrane of rat brain. AB - The effect of GTP on the hydrolysis of [3H]phosphatidylinositol (PI), [3H]phosphatidylinositol-4-phosphate (PIP) and [3H]phosphatidylinositol-4,5 bisphosphate (PIP2) by phospholipase C of rat brain plasma membrane, microsomes and cytosol was determined. Moreover the regulation of PI and PIP phosphorylation by GTP in brain plasma membrane was investigated. In the presence of EGTA PIP2 was actively degraded, opposite to PI and PIP which require Ca2+ for their hydrolysis. Addition of calcium ions in each case caused stimulation of inositide phosphodiesterase(s). GTP independently of calcium ions activates by about 3 times phospholipase C acting on PIP and PIP2 exclusively in the plasma membrane. PI degradation was unaffected by GTP. In the presence of Ca2+ guanine nucleotides have synergistic stimulatory effect on plasma membrane bound phospholipase C acting on PIP2. PIP kinase of brain plasma membrane was stimulated by GTP by about 20-100% in the presence of exogenous and endogenous substrate respectively. PI kinase was negligible activated by about 20% exclusively in the presence of endogenous substrate. These results indicated that guanine nucleotide modulates the level of second messengers as diacylglycerol and IP3 through the activation of phospholipase C acting on PIP2 exclusively in brain plasma membrane. The stimulation of phospholipase C by GTP may occur directly or through the enhancement of substrate level PIP2 due to stimulation of PIP kinase. PMID- 2554173 TI - Cerebellar GABAA receptor binding and function in vitro in two rat lines developed for high and low alcohol sensitivity. AB - The Bmax of the [3H]muscimol binding in the cerebellum of ethanol-naive alcohol sensitive ANT (Alcohol Non-Tolerant) rats was only about 70% of that in the alcohol-insensitive AT (Alcohol Tolerant) rats. There were no line differences in the muscimol binding to cerebrocortical and hippocampal membranes. In the alcohol sensitive rats, the cerebellar [3H]muscimol binding (5 nM) negatively correlated with the ethanol-induced motor-impairment measured in the tilting plane test. Muscimol stimulated the flux of 36Cl- in cerebellar synaptoneurosomes and non filtered microsacs to the same extent in both rat lines. Ethanol produced only a small, although statistically significant, enhancement of the muscimol-stimulated chloride flux in both rat lines. The present data confirms our earlier finding of a low level of muscimol binding in the cerebellar membranes of alcohol-sensitive rats as compared to alcohol-insensitive rats. Further studies are needed to determine the relationship between the Cl- flux stimulation by muscimol and the differential muscimol binding in the cerebellum of these rat lines, and its importance for alcohol sensitivity. PMID- 2554174 TI - Effect of a noncompetitive antagonist (MK-801) of NMDA receptors on convulsions and brain amino acid level in E1 mice. AB - Anticonvulsant action of MK-801, a novel noncompetitive antagonist of N-methyl-D aspartate (NMDA) receptor, was examined in genetically epileptic E1 mice. Systemic injection of MK-801 (0.1-1.0 mg/kg) potently suppressed generalized tonic-clonic convulsions of in a dose-dependent manner (ED50, 0.17 mg/kg). This anticonvulsant effect of MK-801 appeared at a dose which did not induce any obvious behavioral changes. Following the administration of a fully anticonvulsant dose of MK-801 (1 mg/kg), amino acid analysis revealed a significantly elevated level of glycine in the hippocampus. Levels of other amino acids including glutamate, aspartate, taurine, glutamine, alanine, and gamma aminobutyrate were not changed either in the hippocampus or in the cerebral cortex. This study suggests that NMDA system may play an essential role in seizure-triggering mechanisms in E1 mouse. PMID- 2554175 TI - Glucocorticoid and mineralocorticoid receptor mRNA expression in rat brain. AB - In the rat brain, the binding of corticosterone is mediated through two receptor types, the type I receptor and the type II receptor, which are presumed to be encoded by genes designated as MR and GR, respectively. We have studied the regulation of these receptors by glucocorticoids, utilizing a cytosol receptor binding assay. In addition, we have employed molecular probes for the GR and the MR to measure receptor mRNAs. The level of type II receptor binding is uniform across several brain regions, as is the expression of GR (type II) mRNA. In contrast, type I receptor binding is concentrated in the hippocampus, and the MR (type I) mRNA similarly shows a higher level of expression in hippocampus than in the other brain regions studied. Removal of endogenous glucocorticoids by adrenalectomy (ADX) induces an increase, and corticosterone administration results in a decrease, in the level of type I and type II binding in the hippocampus; however, no significant changes in the MR (type I) or GR (type II) mRNA levels are seen with these treatments. The diurnal variation of serum corticosterone in intact rats is correlated with a circadian regulation of type I receptor binding in the hippocampus, while MR (type I) mRNA expression is unaffected. Thus, the changes in type I and type II receptor binding capacity elicited by differing steroid conditions cannot be attributed to modulation of the steady state levels of MR (type I) or GR (type II) mRNA. PMID- 2554176 TI - Effect of 17 beta-estradiol on phosphoinositide metabolism and prolactin secretion in anterior pituitary cells. AB - The present study was undertaken to investigate the effect of 17 beta-estradiol (E2) administration on in vitro prolactin (PRL) release and intracellular phosphoinositide metabolism. The incorporation of [3H]inositol (Ins) into phosphatidylinositol (PtdIns), phosphatidylinositol-4-phosphate [PtdIns(4)P] and phosphatidylinositol-4,5-bisphophate [PtdIns(4,5)P2], and the generation of inositol phosphate (InsPx) following thyrotropin-releasing hormone (TRH) stimulation were studied in primary cultures of anterior pituitary cells obtained from ovariectomized rats. Administration of polyestradiol phosphate (PEP) to ovariectomized rats produced a significant increase (p less than 0.05) in serum PRL levels. This treatment also enhanced significantly (p less than 0.01) the in vitro release of PRL in a progressive manner during 24, 48 and 72 h of culture of dispersed anterior pituitary cells. The radioisotopic labeling by [3H]Ins of all species of phosphoinositides was progressive throughout 72 h of culture, and a good correlation was observed between intracellular phosphoinositide synthesis and PRL release from these cells. PEP treatment enhanced significantly (p less than 0.05-0.01) [3H]Ins incorporation into PtdIns and PtdIns(4)P after 48 and 72 h of culture, although it did not alter [3H]Ins incorporation into PtdIns(4,5)P2. Furthermore, this treatment caused a small, but significant increase (p less than 0.01) in InsPx generation following TRH stimulation. However, the increased [3H]Ins incorporation into phosphoinositide and InsPx generation that we observed after TRH stimulation was significantly (p less than 0.01) less than the increased amount of in vitro PRL release following PEP treatment. There was no significant correlation between the percentage increases in PRL release and phosphoinositide metabolism following the same treatment. These data suggest that phosphoinositide metabolism is enhanced in the anterior pituitary cells of ovariectomized rats by treatment with PEP, but this system does not appear to be tightly coupled or causally related to the much greater production of PRL release. PMID- 2554177 TI - Single-dose ethanol administration activates the hypothalamic-pituitary-adrenal axis: exploration of the mechanism of action. AB - Activation of the hypothalamic-pituitary-adrenal axis (HPAA) by single-dose ethanol administration, which achieved moderately high blood ethanol levels, was explored in naive rats in order to determine the mechanism of ethanol's activation of the stress axis. Adult male rats received a single dose (3.2 g/kg body weight-1 of a 12% solution of ethanol in physiological saline. The plasma concentration of immunoreactive (ir) adrenocorticotropic hormone (ACTH), beta endorphin (BE) and corticosterone (CS) was determined by radioimmunoassay, whereas, plasma concentrations of epinephrine (E) and norepinephrine (NE) were quantified following reverse-phase liquid chromatographic separation and amperometric detection. Ethanol induced maximal plasma ACTH levels within minutes, which declined toward basal levels by 60 min, whereas, plasma concentration of CS rose rapidly and remained elevated at 60 min. Plasma ACTH and CS levels in saline-treated control animals did not vary significantly at any time point. Consistent with co-release of ACTH from corticotrophs, the plasma concentration of ir-BE increased 5-fold at 15 min and declined towards basal levels at 60 min after-ethanol challenge. Plasma E increased 10- to 20-fold as compared to saline controls or preinjection levels and returned to preinjection levels by 90 min, in a manner similar to ethanol-induced changes in proopiomelanocortin-derived peptides and CS. Removal of the adrenal medulla and thus the source of E prior to ethanol administration, did not attenuate activation of the HPAA. Passive immunoneutralization of arginine vasopressin (AVP), using a high-titer AVP antiserum and a protocol which was found to block ether-induced ACTH secretion by 40% in adult male rats, failed to even partially block ethanol-induced ACTH or CS secretion. The results of this study indicate that neither adrenal medulla-derived E nor AVP are significant regulators or coregulators of corticotroph secretions following a moderately high, single-dose, intragastric administration of ethanol. PMID- 2554178 TI - Direct projections from the central amygdaloid nucleus to the hypothalamic paraventricular nucleus: possible role in stress-induced adrenocorticotropin release. AB - The amygdala, particularly the central amygdaloid nucleus, is important for the expression of adrenocorticotropin and corticosterone responses during stress. The aim of the present study was to determine if the central amygdaloid nucleus directly innervated the hypothalamic paraventricular nucleus. To accomplish this aim, the Phaseolus vulgaris leucoagglutinin lectin anterograde tracing method was used. Injections of the tracer into the medial central amygdaloid nucleus resulted in axonal and terminal labeling within the medial and lateral parvocellular parts of the caudal paraventricular nucleus. A dense patch of labeling was observed within the lateral wing of the lateral part of the parvocellular paraventricular nucleus. Only a few labeled axons were observed within the paraventricular nucleus of animals that had lectin injections localized to the lateral part of the central nucleus. Tracer injections localized to the medial amygdaloid nucleus resulted in axonal and terminal labeling primarily within the anterior parvocellular and periventricular regions of the paraventricular hypothalamic nucleus. Sparse to moderate axonal and terminal labeling was observed within the magnocellular parts of the paraventricular nucleus in animals that had injections of tracer into either the medial central nucleus or the medial nucleus. No labeling was observed within the paraventricular nucleus of animals that had injections of lectin within other amygdaloid nuclei or adjacent regions of the striatum. The results demonstrated a topographically organized projection from the amygdala to the hypothalamic paraventricular nucleus. The central nucleus mainly innervates the caudal lateral and medial parvocellular paraventricular nucleus. The medial nucleus innervates the rostral parvocellular parts of the paraventricular nucleus. These pathways could form the anatomical substrates of amygdaloid modulation of neuroendocrine responses to stressors. PMID- 2554179 TI - Multi-dimensional analysis of behavior in mice treated with the delta opioid agonists DADL (D-Ala2-D-Leu5-enkephalin) and DPLPE (D-Pen2-L-Pen5-enkephalin). AB - The effects of intracerebroventricular injection of the delta-selective opioid peptides, DADL (D-Ala2-D-Leu5-enkephalin) and DPLPE (D-Pen2-L-Pen5-enkephalin), on spontaneous locomotor activity were investigated in mice using multi dimensional behavioral analysis, based upon a capacitance system. The analysers classified the movements into 9 sizes (1/1, 1/2, 1/4, 1/8, 1/16, 1/32, 1/64, 1/128 and 1/256). Specific patterns of behavior were each registered on these sizes of movement. At 1.0 and 3.0 micrograms, DADL produced a significant increase in circling (1/4 size of movements) within 15 min after the start of measurements, while it produced a marked increase in linear locomotion (1/2 size), circling (1/4 size), rearing (1/16 size) and grooming (1/32, 1/64 and 1/128 sizes) within 15-30 min after the start. At 10.0 micrograms, DPLPE decreased linear locomotion (1/1 size) and conversely increased circling behavior (1/4 size) within 15 min after the start, whilst this peptide at 3.0 or 10.0 micrograms, produced a marked increase in linear locomotion (1/2 size), circling (1/4 size) and grooming (1/128 size) within 15-30 min after the start. The behavioral effects induced by DADL (3.0 micrograms) and DPLPE (10.0 micrograms) were completely reversed by naloxone (1.0 and 2.0 mg/kg). These results obtained with DPLPE, a delta-selective peptide and DADL, a less delta-selective peptide, indicate a common pattern of activity which was presumably delta receptor mediated. However, one component (linear locomotion, at times immediately after administration of the peptide) did clearly differ between these two peptide analogues.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554180 TI - A local serotonergic component involved in the spinal antinociceptive action of morphine. AB - Participation of opiate, serotonergic and noradrenergic components in the antinociceptive action of intrathecally administered morphine was evaluated by measuring the ability of subcutaneously administered doses of naloxone, methysergide and phentolamine to alter analgesia. Morphine produced a dose dependent elevation of the tail-flick latency, due exclusively to local spinal actions. For example, 10 nmol of the drug, when administered intrathecally in rats with bilateral lesions of the dorsolateral funiculus, produced an increase in the tail-flick latency, that was similar to that observed in intact animals. Furthermore, morphine was ineffective when administered intracerebroventricularly into the fourth ventricle of intact rats. The spinal antinociceptive action of the opiate was antagonized by naloxone (ID50 = 0.035 mg/kg, s.c.) but was also significantly attenuated by methysergide (ID50 = 4.28 mg/kg, s.c.). Phentolamine was ineffective. Doses of methysergide that were most effective in reversing the spinal action of morphine also produced hyperalgesia when administered alone. On the other hand, when the dorsolateral funiculus was lesioned, the hyperalgesia was no longer observed, yet the antagonist remained effective against morphine. These data suggested that the doses of methysergide needed to antagonize the action of morphine were in the same range as those needed to block the synaptic actions of serotonin (5-HT) released from the tonically-acting, descending pain inhibitory nerves. The results demonstrate that local opiate, as well as serotonergic, mechanisms mediate the antinociceptive action of morphine in the spinal cord. The recruitment of a serotonergic component may be related to an action of opiates within the spinal cord, to cause the release of serotonin from the terminal fields of the spinipetal serotonergic nerves. PMID- 2554181 TI - Alpha-adrenergic influences on neuronal responses to visceral afferent input in the nucleus tractus solitarius. AB - Studies were made of the effects of the alpha 1-adrenoceptor agonist methoxamine on spontaneous and synaptically evoked activity in the solitary tract nucleus in isolated, perfused slices of the brain of the rat and those effects were compared to the effects of the alpha 2-agonist clonidine. Methoxamine had no effect on the spike activity of 7 out of 8 spontaneously firing neurones, with no response to electrical stimulation of the solitary tract. Among neurones that responded to tract stimulation, those which were otherwise silent (N = 38) showed a decrease of responsiveness to tract input during the infusion of methoxamine, whereas those with spontaneous activity (N = 10) showed mostly increases of both firing rate and responsiveness. The effects of activation of alpha 2-receptors on responsiveness to tract stimulation were the opposite of previously demonstrated effects of activation of alpha 2-receptors, and these opposing effects could be demonstrated in the same neurone. These results suggest that the transmission of visceral sensory information within the nucleus tractus solitarius may be controlled by opposing alpha 1- and alpha 2-receptor-mediated neural systems. PMID- 2554182 TI - Antagonism of monosynaptic excitations in the mouse olfactory cortex slice by 6,7 dinitroquinoxaline-2,3-dione. AB - The effects of 6,7-dinitroquinoxaline-2,3-dione (DNQX) have been tested in slices of olfactory cortex of the mouse against responses evoked by N-methyl-D aspartate, kainate and quisqualate and on the surface field potentials evoked on electrical stimulation of the lateral olfactory tract. At a concentration of 5 microM, DNQX competitively antagonized responses evoked by kainate and quisqualate, with only a small reduction in the responses to N-methyl-D aspartate. In contrast, DL-(+-)-2-amino-5-phosphonopentanoic acid (APP, 50 microM) selectively antagonized depolarizations to N-methyl-D-aspartate. The amplitude of the field potential known as the N-wave was reduced by DNQX in a concentration-dependent reversible manner (IC50 = 2.92 +/- 0.33 microM; mean +/- SE mean, n = 4). DL-(+-)-2-Amino-5-phosphonopentanoic acid (50 microM) did not significantly affect this action of DNQX. It is concluded that DNQX inhibits monosynaptic excitations in the olfactory cortex by selectively blocking kainate and/or quisqualate receptors, although it is unclear whether the receptors are located at pre- and/or postsynaptic sites. PMID- 2554183 TI - Methamphetamine-induced neuronal damage: a possible role for free radicals. AB - The hypothesis that methamphetamine-induced neuronal damage is mediated by the production of free radicals was evaluated by pretreating rats with either antioxidants or a superoxide dismutase (SOD) inhibitor. It was found that methamphetamine (dose range 6.25-25.0 mg/kg) caused long-lasting depletions of dopamine and serotonin in the striatum and that pretreatment with the antioxidants, ascorbic acid (10-100 mg/kg), ethanol (1 g/kg), mannitol (2 g/kg), or vitamin E (2 g/kg), attenuated these depletions, whereas pretreatment with the superoxide dismutase inhibitor diethyldithiocarbamate (200-400 mg/kg) exacerbated the depletions. The alteration of this effect by four different antioxidants, as well as an inhibitor of superoxidase dismutase, indicated that oxygen-free radicals may have a role in the methamphetamine-induced neurotoxicity. PMID- 2554185 TI - Gamma-aminobutyric acid and benzodiazepine receptors in cultured cerebellar granule cells: effects of taurine and its lipophilic derivatives. AB - The effects of taurine and some lipophilic derivatives of taurine on binding to GABA and benzodiazepine receptors were studied in intact cerebellar granule cells. The phenylsuccinylimido derivatives of taurine appeared to increase the binding of muscimol in micromolar concentrations, while taurine decreased it slightly. Only minor changes were seen in the basal binding of flunitrazepam, whereas stimulation of the binding by GABA was strongly reduced by piperidino, benzamido and phenyl-succinylimidotaurine with taurine itself again showing only a weak effect. Diphenylhydantoin, which bears structural resemblance to the phenylsuccinylimido group, had a strong effect on the stimulated binding of flunitrazepam and it also slightly reduced the basal level of binding. Thus, it seems possible that the effects of the phenylsuccinylimido derivatives of taurine on the binding of flunitrazepam were due to this chemical structure and not to the taurine-like core of the molecules. The phthalimido derivative of taurine, taltrimide, which has been tested in clinical trials with epileptic patients, did not show any activity in the binding studies. PMID- 2554184 TI - Benzodiazepine receptor autoradiography in corpus striatum of rat after large frontal cortex lesions and chronic treatment with diazepam. AB - It has previously been shown that diazepam impairs behavioural recovery from partial unilateral ablation of the cerebral cortex in rats. The present study confirmed this in rats with large unilateral lesions of the frontal cortex and showed that diazepam (5 mg/kg i.p. daily for 14 days immediately after surgery) prevented recovery from sensory asymmetry even after 120 days. In saline-treated rats greater than 80% recovery had occurred by this time. A study of binding to benzodiazepine receptors, using an in vitro autoradiographic technique, was performed to determine whether the lack of recovery after administration of diazepam was associated with any long-term receptor changes on the damaged side of the brain. Binding of [3H]Ro15-1788 was increased by up to 40% in the caudate putamen on the decorticated side at 14-120 days. This was not significantly altered by treatment with diazepam. Binding of [3H]Ro15-1788 in the nucleus accumbens was not altered by lesion of the frontal cortex alone or after treatment with diazepam. It is concluded that the lack of recovery from sensory asymmetry, produced by diazepam after lesion of the frontal cortex cannot be correlated with any change in binding to benzodiazepine receptors within the corpus striatum. PMID- 2554186 TI - Potentiation of apomorphine-induced stereotyped behaviour by acute treatment with dopamine depleting agents: a potential role for an increased stimulation of D1 dopamine receptors. AB - Treatment of mice with reserpine (10 mg/kg, s.c.) and alpha-methylparatyrosine (400 mg/kg) led to the potentiation of stereotyped behaviour, induced by apomorphine (0.37-1.5 mg/kg, s.c.), i.e. to the appearance of licking and gnawing in addition to climbing and sniffing occurring in control mice. Similar results were obtained by combined treatment with SK&F 38393 (30 mg/kg, s.c.) and RU 24926 (15 mg/kg, i.p.). In mice treated with dopamine depleting agents, SCH 23390 (1.25 20 micrograms/kg, s.c.) and metoclopramide (0.62-20 mg/kg, i.p.) antagonized gnawing induced by 0.75 mg/kg (s.c.) apomorphine, at doses significantly larger than those required for the antagonism of climbing and sniffing. The same treatment with reserpine and alpha-methylparatyrosine produced an increased formation of cyclic AMP, induced by SK&F 38393 (10(-8)-10(-4) M), from homogenates of the striatum of the rat. Potentiation of apomorphine-induced stereotyped behaviour and increased SK&F 38393-induced formation of cyclic AMP had similar time-courses with a maximum 18 hr after treatment. These data suggest that the potentiation of apomorphine-induced stereotyped behaviour produced by acute treatment with dopamine depleting agents is at least partly due to an increased activity of the adenylate cyclase linked to D1 dopamine receptors. Finally, a small dose of amisulpride (a discriminant benzamide derivative) potentiated the stereotyped behaviour induced by the combined treatment with SK&F 38393 and RU 24926 in naive mice and, in a more marked manner, in mice treated with dopamine depleting agents; amisulpride did not produce stereotyped behaviour when combined with SK&F 38393 or RU 24926 administered alone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554187 TI - Changes in multiple opioid receptors in regions of the brain in rats treated chronically with thyroxine. AB - The effect of the administration of thyroxine (T4) (1 mg/kg, s.c.), to male Sprague-Dawley rats on alternate days for 18 days, on the binding of opioid ligands, [3H]Tyr-D-Ala-Gly-MePhe-Gly-ol (DAGO, mu receptors), [3H]Tyr-D-Ser-Gly Phe-Leu-Thr (DSTLE, delta receptors) and [3H]ethylketocyclazocine (EKC, kappa receptors) to membranes of regions of the brain was determined. The chronic administration of thyroxine to rats decreased their rate of gain of body weight, increased colonic temperature and increased systolic blood pressure and heart rate, in comparison to vehicle-injected rats. The administration of thyroxine also increased the serum concentration of triiodothyronine (total T3) and thyroxine, when compared to vehicle-injected rats. The binding of [3H]DAGO to membranes of the striatum of rats treated with thyroxine was greater than in vehicle-treated rats; however, the binding to membranes of pons and medulla, amygdala, hypothalamus, midbrain and cortex in the two treatment groups did not differ. The increased binding of [3H]DAGO in rats treated with thyroxine was due to an increase in the Bmax value. The binding of [3H]DSTLE in the midbrain, hypothalamus, pons and medulla and striatum of rat treated with thyroxine and vehicle-injected rats did not differ but it was significantly less in the amygdala of rats injected with thyroxine than in vehicle-injected rats. The decreased binding in the amygdala was due to changes in the Kd value of [3H]DSTLE.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554188 TI - Effects of cocaine on neuromuscular transmission in the lobster. AB - In vitro electrophysiological techniques were used on an excitatory neuromuscular junction of a walking-limb stretcher muscle of the lobster in order to define the pharmacology of cocaine on a glutamatergic synapse. Cocaine reduced the amplitudes of the excitatory and the miniature junction potentials, as well as the mean quantum content of the muscle fiber. The effect on mean quantum content points to a presynaptic site of action of the drug, and the effect on miniature junctional potential suggests a postsynaptic site of action of the drug. In addition, cocaine shifted the concentration-depolarization curve for glutamate to the right, demonstrating a postsynaptic depression of the response to glutamate. Cocaine exerted no effect on the resting membrane potential or the effective membrane resistance of the muscle fibers. The data indicate that the drug concomitantly exerted pre- and postsynaptic depression of a glutamatergic junction. PMID- 2554189 TI - Long-lasting inhibitory action of caerulein on climbing fiber system in the cerebellum of the rat. AB - Caerulein caused a marked decrease in levels of guanosine 3',5'-cyclic monophosphate (cGMP) in the cerebellum in rats. This effect was observed to be dose-dependent after the intraperitoneal administration of caerulein for doses over 20 micrograms/kg and lasted for about 4 hr in doses of 100 micrograms/kg. However, in vagotomized rats, caerulein failed to alter the level of cGMP in the cerebellum. Caerulein suppressed harmaline-induced increases in cGMP in the cerebellum for more than 30 hr. In contrast, the increases in levels of cGMP in the cerebellum, induced by treatment with methamphetamine, apomorphine and picrotoxin, were not inhibited by pretreatment with caerulein. These results suggest that the peripheral administration of caerulein can inhibit the activity of climbing fibers for a long period of time in the cerebellum of the rat through the stimulation of the abdominal vagus nerves. PMID- 2554190 TI - Opioid systems in the amygdala can serve as substrate for the behavioral effects of the ACTH-(4-9) analog ORG 2766. AB - Rats housed individually for 7 days showed a marked decrease in motor activity when tested under intense light conditions in a novel environment as compared to group-housed rats tested under low light conditions. The ACTH analogue ORG 2766 administered into the amygdala decreased the motor activity of group-housed rats tested under low light conditions and increased the motor activity of 7-days isolated rats tested under intense light conditions (ED50: 1-10 pg). Injection of the peptide into the nucleus accumbens was not effective, suggesting that ORG 2766 affects the integration of sensoric stimuli rather than the specific motor output systems. Pretreatment of the rats with the opiate antagonist naltrexone in the amygdala completely blocked the effect of ORG 2766. A similar blockade of the ORG 2766-induced effect could be induced by pretreatment with endorphin antibody suggesting that the "normalizing" activity of ORG 2766 on environmentally induced behavioral changes is mediated by the release of endogenous opioid peptides. PMID- 2554191 TI - Tolerance develops to the behavioural effects of ACTH-(1-24) during continuous i.c.v. infusion in rats, and is associated with increased hypothalamic levels of beta-endorphin. AB - In rats, the continuous infusion of ACTH-(1-24) into a brain lateral ventricle (0.5 micrograms/h in the volume of 1.11 microliters, for 7 days) caused a significant inhibition of the subsequent behavioural response to the acute intracerebroventricular injection of the same peptide. Tolerance developed to all the most typical signs of the ACTH-induced behavioural syndrome (grooming, stretching, yawning, penile erection, inhibition of food intake), and was associated with a significant increase in the hypothalamic levels of beta endorphin immunoreactivity. PMID- 2554192 TI - Investigations with the specific mu-opiate receptor agonist fentanyl in depressive patients: growth hormone, prolactin, cortisol, noradrenaline and euphoric responses. AB - 15 depressive patients and 15 controls, 9 of them age- and sex-matched, were administered 0.2 mg/70 kg i.v. fentanyl, a specific and highly potent mu-opiate receptor agonist. Growth hormone response was significantly reduced in depressive patients in comparison to controls, whereas prolactin response did not significantly differ between the two groups. Cortisol plasma concentration increased in depressive patients and decreased in controls. The difference between the groups failed to reach statistical significance. Only in patients, but not in controls, fentanyl led to a significant increase in plasma noradrenaline. In contrast, a significant increase in the visual analogue scale for the evaluation of psychotropic drug effects was found only in controls after fentanyl administration. From these preliminary results in connection with other studies we conclude a possible involvement of a disturbed opioid system at least in a subgroup of depressive patients. PMID- 2554193 TI - Further investigation on benzodiazepine binding inhibitory activity in patients with major depression or panic disorder and in healthy volunteers. AB - Preliminary findings regarding the presence of plasmatic benzodiazepine binding inhibitory activity (BBIA) in 14 psychiatric patients prompted us to investigate it further in larger samples of psychiatric patients (n = 44) and in healthy controls (n = 14). The results have shown that BBIA is present in all the subjects included with statistically significant differences between the patients and controls. The highest concentrations were found in the patients with no difference between anxious or depressed patients, and the lowest in the healthy controls. These findings indicate that BBIA might play a role in the pathophysiology of some manifestations of anxiety. PMID- 2554194 TI - Effect of hydrocortisone on synaptic transmission in surviving rat olfactory cortex slices. AB - A study has been made of the effect of bath-applied hydrocortisone (10(-4) M) on excitatory transmission of the lateral olfactory tract (LOT), a superficial pyramidal cell synapse of the rat olfactory cortex slice. Populations EPSP and IPSP were depressed for 6-12 min and were then increased (60 min) by hydrocortisone. Recovery of evoked potentials was prolonged (40-60 min) when slices were washed with normal incubation medium. Habituation of synaptic potential during LOT repeated stimulation deteriorated as a result of hydrocortisone application in perfusion solution. The data obtained were considered to represent functions of adaption of the nervous system to a stressor. PMID- 2554195 TI - Trochanteric sciatic neuropathy. AB - A patient with severe weakness, atrophy, and sensory loss of the right leg had a focal right sciatic neuropathy. The sciatic nerve was enlarged at the level of the lesser trochanter, excessively firm, and multistranded; its stimulation threshold was focally increased. Biopsied fascicles had reduplicated perineurial leaflets, many Renaut bodies, and an abnormal unimodal spectrum of small-diameter fibers. We postulate that the lesion was induced by the combination of an underlying prominent lesser trochanter and sitting on hard benches. PMID- 2554197 TI - Familial adenomatous polyposis. Definition, description and genetics. PMID- 2554196 TI - [Familial polyposis: epidemiology]. PMID- 2554198 TI - [Role of endoscopy in familial polyposis: diagnosis and therapy]. PMID- 2554199 TI - Familial adenomatous polyposis: surgical strategy. PMID- 2554200 TI - Restorative proctocolectomy for familial adenomatous polyposis. PMID- 2554201 TI - Extracolonic manifestations of familial polyposis coli. PMID- 2554202 TI - [The Registry of Familial Colonic Polyposis and its implementation]. PMID- 2554203 TI - [Proctocolectomy and ileo-anal reservoir in the treatment of ulcerous rectocolitis and diffuse colonic polyposis. A report on the Italian registry and on a new model of the 3-loop reservoir]. PMID- 2554204 TI - Phenotypic and functional analysis of circulating phagocytic cells in IgA nephropathy patients. AB - We analysed the possible mechanisms responsible for the functional abnormalities reported on phagocyte cell function in IgA nephropathy patients. The oxidative metabolism by the superoxide anion ferricytochrome C reduction test on monocytes and the chemiluminescence response on neutrophils was examined in 32 patients. Moreover the expression of cell membrane structures involved in immune response and phagocytosis (i.e. Fc receptors and HLA class II antigens) was evaluated cytofluorimetrically using specific monoclonal antibodies. Our own results did not show striking differences in oxidative potential and phenotype of patients' phagocyte cells as compared with healthy controls. However a HLA class II associated structure was found to be quantitatively more expressed in patients' monocytes than in normal controls (P less than 0.005). These findings suggest that the defective phagocyte system function reported in IgA nephropathy patients is not simply due to a loss of cell surface receptors, nor to abnormalities in intracellular metabolic pathways implying oxygen consumption. The increase in expression of some HLA class II structures, often associated with the presence of circulating IgA immune complexes, probably reflects a protracted immunological stimulation. PMID- 2554205 TI - Na+ channel accumulation on axolemma of afferent endings in nerve end neuromas in Apteronotus. AB - In mammals, cut sensory axons trapped in a nerve end neuroma have been shown to develop hyperexcitability, and to become a source of ectopic afferent discharge and abnormal sensation. We have explored cellular mechanisms underlying neuroma electrogenesis. First we confirmed that ectopic neuroma discharge develops in injured afferents in the electrosensory lateral line nerve of the weakly electric fish Apteronotus, as it does in mammals. Then, using previously characterized antibodies that specifically recognize Na+ channel proteins in this species, we obtained light and electron microscopic evidence of abnormally intense immunolabelling of axolemma at the injury site. Accumulation of excess Na+ channels in afferent endings in neuromas could account for their electrical hyperexcitability. PMID- 2554206 TI - Hippocampal synaptic plasticity induced by excitatory amino acids includes changes in sensitivity to the calcium channel blocker, omega-conotoxin. AB - The hippocampus is widely used in investigations of different forms of synaptic plasticity, including long-term potentiation and kindling. Receptors for excitatory amino acids (EAAs) play a prominent role in these phenomena. Recently, is has been demonstrated that exposure of hippocampal slices to EAAs and related agonists produces biphasic effects on excitatory synaptic transmission: initial blockade of synaptic responses is followed by a delayed recovery. The recovered responses demonstrate altered pharmacological properties: they acquire sensitivity to N-methyl-D-aspartate (NMDA) antagonists during L-glutamate (Glu) exposure and lose sensitivity to both NMDA and non-NMDA antagonists under L aspartate (Asp). These changes persist for many hours. It was suggested that this form of hippocampal plasticity may involve transitions between distinct states of synaptic functioning. To explore this possibility, we investigated several properties of synaptic transmission in the initial and EAA-modified states. Here we report that hippocampal postsynaptic potentials (PSPs) evoked under Glu or Asp exposure completely lose sensitivity to omega-conotoxin GVIA (omega-CgTX), a potent, specific, and irreversible blocker of certain types of neuronal calcium channels. After washout of the EAA, sensitivity to the toxin is regained. These results indicate that prolonged EAA exposure induces profound changes in the machinery of synaptic transmission, which include, but are not limited to, changes in calcium channel functioning. PMID- 2554207 TI - Electrophysiological evidence for receptors for vasoactive intestinal peptide and angiotensin II on astrocytes of cultured rat central nervous system. AB - Electrophysiological studies have been made of the actions of vasoactive intestinal peptide (VIP) and angiotensin II (Ang II) on astrocytes in explant cultures of rat brain stem and spinal cord. Addition of VIP to the bathing solution at concentrations of 10(-8) to 10(-6) M caused hyperpolarizations. Ang II at the same concentrations depolarized the majority of astrocytes tested. At 10(-9) M, both peptides had no action or only small effects. In approximately one third of the cells, the depolarizations by Ang II were accompanied by rhythmic oscillations. The Ang II antagonist saralasin reversibly blocked the depolarizations by Ang II. These findings, together with autoradiographic binding studies from our laboratory strongly suggest the existence of VIP and Ang II receptors on astrocytes. PMID- 2554208 TI - Postnatal development of the spinal cord thyrotropin-releasing hormone receptor in male and female rats. AB - The density of the spinal cord thyrotropin-releasing hormone (TRH) receptor in male and female rats was measured during postnatal development. The younger rats of both sexes (11-16 days old) exhibited higher levels of TRH-binding than did adults. A comparison of the properties of receptor binding for young and adult spinal cords showed that the increased binding resulted solely from an increase in maximum binding capacity rather than a change in receptor affinity. Furthermore, no sex-dependent difference in the receptor property was found at any stage of development. In conclusion, these findings demonstrate that the differences in the maturation of TRH receptor itself may not be the cause of known gender-specific differences in the physiologic actions of TRH in the spinal cord of young rats. PMID- 2554209 TI - Microiontophoresis of flurazepam on inspiratory and postinspiratory neurons in the ventrolateral medulla of cats: an intracellular study in vivo. AB - Effects of flurazepam on the periodic inhibitory postsynaptic potentials (IPSPs) and on the action of locally applied gamma-aminobutyric acid (GABA) were studied in bulbar respiratory neurons of decerebrate cats using concentric multibarrelled electrodes for intracellular recording and extracellular iontophoresis. Iontophoresis of flurazepam augmented spontaneous IPSPs and increased the hyperpolarization induced by GABA. Iontophoretic application of bicuculline suppressed the action of flurazepam. The reversal potential for spontaneous IPSPs was similar to that for the GABA-response. Intracellular Cl- injection shifted both the IPSP wave and the GABA response in a depolarizing direction. Flurazepam enhanced these depolarizing responses. These results suggest that GABA mediates the postsynaptic inhibition in bulbar respiratory neurons. PMID- 2554210 TI - Acute excitotoxicity in chick retina caused by the unusual amino acids BOAA and BMAA: effects of MK-801 and kynurenate. AB - beta-N-Oxalylamino-L-alanine (BOAA) and beta-N-methylamino-L-alanine (BMAA) were tested for their ability to produce acute excitotoxicity in in embryonic chick retina. gamma-Aminobutyric acid (GABA) release and histology were monitored in retina treated with various concentrations of BOAA, BMAA, kainate (KA), N-methyl D-aspartate (NMDA), or glutamate. BOAA and BMAA caused retinal lesions similar to those produced by the excitatory amino acids. BOAA was slightly less potent than KA, whereas BMAA had a potency similar to glutamate. BOAA, like KA and NMDA, caused a dose-dependent increase in GABA release. Addition of the NMDA antagonist (+)-MK-801, completely blocked acute excitotoxicity caused by NMDA or BMAA but was ineffective against KA or BOAA. Kynurenate, a nonspecific glutamate receptor antagonist, and DIDS, a Cl- channel blocker, were effective in blocking all agonist-induced toxicity. It is concluded that BOAA and BMAA cause excitotoxic damage in retina; BOAA induces toxicity through a non-NMDA type glutamate receptor and BMAA through the NMDA receptor. PMID- 2554211 TI - Effects of metaphit on phencyclidine and serotonin2 receptors. AB - We have investigated whether metaphit, a derivative of phencyclidine (PCP) which irreversibly binds to a population of PCP receptor sites in rat brain, blocks PCP induced head-twitch response which is produced through serotonin2 (5-HT2) receptors, and also whether metaphit decreases the capacity of 5-HT2 receptors. Metaphit (1 mumol/rat) had decreased the intensity of PCP-induced head-twitch response and had depleted both PCP and 5-HT2 receptors by 24 h after administration, but it failed to block 5-HT agonist 5-methoxy-N,N dimethyltryptamine-induced 5-HT1A receptor-dependent behaviors. These results reconfirmed our hypothesis that PCP and 5-HT2 receptors may have very similar binding sites. PMID- 2554212 TI - Superoxide dismutase treatment reduces [3H]flunitrazepam affinity in cortex and hippocampus of the rat. AB - Superoxide dismutases (SOD) are essential enzymes involved in the cellular defense against oxidative processes occurring with the generation of the superoxide anion. In this work, we have investigated in the rat the effects of a subchronic administration of liposomal SOD from bovine erythrocytes, on benzodiazepine (BZD) 'central-type' receptor sites in cortex and hippocampus. Animals were treated for 15 days with an i.p. injection of liposomal SOD, and binding parameters were determined using [3H]flunitrazepam. BZD receptor affinity was found decreased, while no change was observed in the maximal binding capacity. With regard to previously reported data, our results show that the superoxide anion radical is involved in the modulation and/or the stability of 'central-type' BZD receptor sites. PMID- 2554214 TI - Chronic thyrotropin-releasing hormone decreases the affinity and increases the number of its own receptor in the spinal cord. AB - Effect of chronic thyrotropin-releasing hormone (TRH) treatment on the properties of spinal cord TRH-receptor was evaluated by Scatchard analysis of data on the saturation of specific binding of [methyl-His3-3H]TRH. Continuous subcutaneous administration of TRH (0.51 +/- 0.02 mg/kg/day) via an Alzetmini osmotic pump for ten days led to a substantial increase in both Bmax (control: 20.0 +/- 2.9, treated: 36.1 +/- 2.8 fmol/mg protein, P = 0.0018) and Kd (control: 7.6 +/- 0.8 treated: 11.7 +/- 1.8 nM, P = 0.0349) of the binding sites. These data suggest that chronic TRH treatment may not induce desensitization of physiologic/pharmacologic responses of TRH in the spinal cord. PMID- 2554213 TI - Ubiquitin gene expression in brain and spinal cord in motor neurone disease. AB - A restriction fragment of the coding region of a human ubiquitin gene has been used in Northern analyses of RNA prepared from human motor cortex and anterior horn region of cervical spinal cord. The analyses show that there is a substantial increase (approximately two-fold) in the expression of a polyubiquitin gene in motor cortex and spinal cord from patients with motor neurone disease compared to these tissues from control cases. Polyubiquitin gene expression in other organisms is associated with physical or chemical cell stresses. The data indicate that the primary stresses which result in the generation of ubiquitinated filamentous inclusion bodies in neurones in motor neurone disease also result in increased transcription of a gene coding for a polyprotein of ubiquitin. PMID- 2554215 TI - Effects of either forskolin, the 1,9-dideoxy derivative of forskolin, or 8 bromocyclic AMP on cyclic AMP and melatonin production in the Syrian hamster pineal gland in organ culture. AB - The possible involvement of cyclic AMP in the regulation of melatonin production was investigated in cultured Syrian hamster pineal glands. Addition of forskolin, an adenylate cyclase activator, to the incubation medium caused marked increases in both cyclic AMP and melatonin levels in glands collected in the second half of the dark period. 8-Bromocyclic AMP, an analogue of cyclic AMP, also increased melatonin production. The 1,9-dideoxy derivative of forskolin, which is unable to activate the cyclase, was ineffective in stimulating either cyclic AMP levels or melatonin production. These results support a primary role for cyclic AMP in the nocturnal increase of melatonin production in the Syrian hamster pineal gland. PMID- 2554216 TI - L-type calcium channels in the regulation of neurite outgrowth from rat dorsal root ganglion neurons in culture. AB - Dorsal root ganglion neurons from neonatal rats were grown in culture for 12 h and the extent of neurite outgrowth determined by counting the fraction of neurons with neurites. In the presence of high K+, veratridine or bradykinin the extent of neurite outgrowth was reduced by about 60%. The inhibitory effect of depolarisation was reversible and was abolished by nifedipine. gamma-Aminobutyric acid (GABA), baclofen and 2-chloroadenosine had no effect on neurite outgrowth in control cultures but abolished the inhibitory effect of depolarisation. PMID- 2554217 TI - alpha-Difluoromethylornithine (DFMO) disturbed the sensorimotor functional recovery from a sciatic lesion. AB - The effects of inhibition of polyamine synthesis on the motor and sensory functional neural recovery after a sciatic crush lesion was studied by measuring sensory and motor function of a crushed sciatic nerve after daily exposure to alpha-difluoromethylornithine (DFMO). DFMO increased the time needed for functional motor recovery by 13% in the toe-spread test when the lesion was about 20 mm proximal to the target muscle. DFMO reduced the rate of sensory axonal elongation by 24% in the pinch-test. PMID- 2554218 TI - Transient increase in ligand binding to quisqualate and kainate sites in cerebral cortex of immature rats. AB - The postnatal changes in the specific binding of [3H]kainate and [3H]AMPA (RS alpha-amino-3-hydroxy-5-methyl-4-isoxasolopropionic acid, an agonist of quisqualate receptors) were studied in cerebral cortex of rats, aged 2-360 days. The binding of the two ligands was assayed in whole-tissue homogenates. Similar developmental time courses were found for kainate and AMPA binding, characterized by high perinatal levels, a further increase during the first few days after birth, an early maximum value around the age of one week, and a gradual decrease to adult values which were attained at an age of 3-4 weeks. As revealed by Scatchard analysis, the transient elevation of ligand binding was derived from an increased density of binding sites, which, in the case of AMPA, was accompanied also by an increase in binding affinity. The results indicate that, in the immature cerebral cortex, kainate and quisqualate receptors may play a role other than in synaptic transmission. PMID- 2554219 TI - Differential affinities of TRH analogs at the mammalian spinal cord TRH receptor: implications for therapy in spinal injuries. AB - The equilibrium receptor binding properties and the pharmacological specificity of the spinal receptors for thyrotropin-releasing hormone (TRH) were determined. [3H]MeTRH bound to a single class of high-affinity (dissociation constant, KdS = 5.0; 6.0 and 5.5 nM), saturable (Bmax = 21.5, 32.6 and 130.7 fmol/mg protein) binding sites for TRH in homogenates of the rat, guinea pig and rabbit spinal cord respectively. [3H]MeTRH receptor binding was competitively but differentially inhibited by TRH analogs. The inhibition constants (Kis) in the three spinal cord preparations were: MeTRH (4.2-5.9 nM); TRH (14.2-33.9 nM); RX77368 (113-122 nM); CG3703 (117-142 nM); MK-771 (122-140 nM); CG3509 (10-32 microM); NVal2-TRH (32-56 microM) and TRH free acid (37-73 microM). These data have shown that the parent tripeptide. TRH, and its methylated analog. MeTRH, are the most potent displacers of [3H]MeTRH receptor binding, and that N-terminus modifications (as in CG3703, CG3509). C-terminus modifications (as in RX77368, TRH free acid) and both N- and C-terminus modifications (as in MK-771) of TRH result in markedly reduced affinity for the TRH receptor. Although, CG3703 and CG3509 have been previously found to be almost equally effective in the treatment of spinal cord injury, we have found that CG3703 has a significantly higher (70- to 283-fold) affinity than CG3509 for spinal TRH receptors. Interestingly, although RX77368 and MK-771 appear to have similar TRH receptor affinities to CG3703, the former analogs have shown less beneficial effects in animal models of spinal injury.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554220 TI - The specific NMDA receptor antagonist AP-7 attenuates focal ischemic brain injury. AB - Pharmacologic blockade of excitatory amino acid receptors, especially the N methyl-D-aspartate-preferring subclass of glutamate receptors, has been shown to reduce neuronal damage in models of global cerebral ischemia followed by reperfusion. The pharmacologic blockade at the NMDA receptor attenuates infarct size following permanent focal vascular occlusion in brain. Functional recovery is improved as well. These effects were seen with treatment begun 15 min following the stroke. PMID- 2554221 TI - Hippocampal CA1 pyramidal cells of rats have four voltage-dependent calcium conductances. AB - In isolated rat hippocampal neurons, we observed 4 voltage- and extracellular Ca2+-dependent conductances; i.e. the T-, N- and L-type Ca2+ currents and tetrodotoxin-sensitive transient Ca2+ current. Intracellular perfusion with F- suppressed irreversibly the L-type Ca2+ current and partially the N-type one. omega-Conotoxin inhibited selectively the L-type Ca2+ current. Amiloride reduced strongly the T-type Ca2+ current without affecting the L-type one. Gd3+, nicardipine, phenytoin and octanol had no specific inhibition on the T-, N- and L type Ca2+ currents. Thereby, the pharmacological property of mammalian CNS neurons for Ca2+ channel blockers considerably differs from that in the peripheral and cultured cells. PMID- 2554222 TI - The EPSP-spike (E-S) component of long-term potentiation in the rat hippocampal slice is modulated by GABAergic but not cholinergic mechanisms. AB - Long-term potentiation of synaptic efficacy (LTP) can be shown to consist of two components: a synaptic and an excitatory postsynaptic potential (EPSP)-spike (E S) component. The E-S component is expressed as a leftward shift in the curve relating population spike amplitude as a function of EPSP slope. The participation of cholinergic and GABAergic processes in E-S potentiation was studied in field CA1 of rat hippocampal slices. Atropine, a muscarinic antagonist, did not prevent tetanus-induced E-S potentiation. The cholinergic agonist carbachol and the GABAA antagonist picrotoxin produced a leftward shift in the E-S relation; picrotoxin, but not carbachol, prevented the expression of tetanus-induced E-S potentiation. These observations indicate that an increase in the ratio of evoked excitation to inhibition and/or a reduction in tonic inhibition mediated by the activation of GABAA receptors contribute to E-S potentiation produced by high-frequency stimulation. PMID- 2554223 TI - gamma-Aminobutyric acid elevates cytosolic Ca in bovine chromaffin cells. AB - Measurements of the cytosolic Ca concentration ([Ca]i) with the Ca-sensitive dye, fura-2, showed that in intact, but not in voltage-clamped, bovine chromaffin cells gamma-aminobutyric acid (GABA, 10 microM) elicited a transient increase in [Ca]i. The Ca transient of intact cells was inhibited by bicuculline (20 microM), by removal of extracellular Ca or by treatment with the Ca channel blocker cobalt (2.5 mM), and enhanced by lowering the extracellular Cl. We conclude, that GABA elevates [Ca]i by inducing a GABAA-receptor-linked Cl current which depolarizes the cell membrane sufficiently to activate potential-operated Ca channels and cause Ca entry into the cell. PMID- 2554224 TI - Excitatory amino acid receptors mediate slow synaptic transmission in turtle cerebellum. AB - In the isolated turtle cerebellum intracellular recordings from Purkinje cell dendrites and somata reveal novel slow excitatory synaptic potentials evoked by activation of climbing fiber (CF) or parallel fiber (PF) inputs. Classical fast excitatory synaptic responses to CF and PF stimulation are followed by large, slow excitatory postsynaptic potentials (sEPSPs) which are associated with an increase in conductance and are enhanced by hyperpolarization. Both sEPSPs are blocked by the excitatory amino acid (EAA) antagonist kynurenate, but not by DL-2 amino-5-phosphonovalerate (AP-5). The EAA receptor antagonist L-amino-4 phosphonobutyric acid (L-AP-4) reversibly blocked the PF-sEPSP without affecting the CF-sEPSP. Two novel slow synaptic potentials mediated by excitatory amino acid receptors can therefore be observed in turtle cerebellum which may play an important role in synaptic integration. PMID- 2554225 TI - Determination of GABA receptor-linked Cl- fluxes in rat cerebellar granule cells using a fluorescent probe SPQ. AB - Gamma-Aminobutyric acid (GABA)-induced Cl- fluxes in cultured rat cerebellar granule cells were measured using the chloride-sensitive fluorescent probe SPQ (6 methoxy-N-(3-sulphopropyl)quinolinium) incorporated into the cells. The fluorescence of SPQ is quenched by Cl-ions. GABA and pentobarbitone increased the fluorescence of the probe when the Cl- gradient was directed outward by bathing cells, grown in the presence of GABA, in a low Cl- medium. Picrotoxin and bicuculline inhibited the response to GABA. The results suggest that SPQ is a suitable probe for measuring GABA-induced Cl- fluxes in living cells. PMID- 2554226 TI - The molluscan neuropeptide FMRFamide stimulates the release of [14C]acetylcholine from isolated ileal synaptosomal preparations of guinea-pig. AB - FMRFamide stimulated, in a dose-dependent manner, the efflux of [14C]acetylcholine (ACh) from isolated synaptosomes of guinea-pig ileum preloaded with labelled choline. Participation of the cholinergic mechanism and/or substance P was ruled out by the finding that antagonists failed to affect the FMRFamide-induced release of ACh. The ACh-releasing action of FMRFamide was negated by the deletion of calcium ion from the bathing medium and it was also abolished by tetrodotoxin. The results obtained suggest that FMRFamide possesses the ability to induce the release of ACh from enteric synaptosomes of guinea-pig. PMID- 2554227 TI - Pertussis (whooping cough) toxin and Bordetella pertussis whole-cell antibody levels in a healthy New Zealand population. AB - Enzyme linked immunosorbent assay (ELISA) tests were used to measure IgG antibody levels to Bordetella pertussis whole-cell and to pertussis toxin in a healthy New Zealand population. The percentage of individuals with measurable antibody to B pertussis whole-cell increased from 47.3% in the 5 year olds to 93.0% in the 40 49 age group, from which the percentage with antibody dropped to 88.7% in the 50 65 age group. The percentage of persons with antibody to the toxin increased with age from 17.5% in the 5 year olds to 67.0% in the 40-49 age group, from which the percentage of individuals with antibody dropped to 45.4% in the 50-65 age group. Similarly, the percentage of persons with antibody to both B pertussis whole-cell and pertussis toxin increased with age from 15.5% in the 5 year olds to 63.0% in the 40-49 age group, and then the percentage with antibody to whole-cell and toxin dropped to 45.4% in the 50-65 age group. The percentage of individuals with antibody to pertussis toxin is generally markedly lower than the percentage of persons with antibody to B pertussis whole-cell across the age range. Suggestions are made as to how to improve herd immunity for pertussis. PMID- 2554228 TI - Human papillomavirus genotype as a prognostic indicator in carcinoma of the uterine cervix. AB - The clinical implications of specific human papillomavirus (HPV) types in invasive cervical carcinomas are only beginning to be appreciated. In this series of 100 women with cervical cancers analyzed for the presence of HPVs 6, 11, 16, 18, and 31 by Southern blot hybridization, a more aggressive clinical behavior was demonstrated for tumors containing HPV 18 than for those with HPV 16 or those in which no HPV was identified. Among 69 stage Ib tumors, no significant differences were found in the size of tumor, presence of parametrial involvement, or lymph node metastasis among patients whose tumor contained HPV 16, HPV 18, or no HPV DNA; however, 45% of the women with HPV 18-containing tumors (five of 11) had recurrence, as compared with only 16% of those with HPV 16 (five of 31) during the 20-month mean follow-up period. This tendency for HPV 18-containing tumors to recur was seen with all histologic subtypes of cervical cancers and with all grades of tumor. In addition, patients with HPV 18-containing tumors were more likely to give a history of recent normal Papanicolaou smears than were those whose tumors contained HPV 16. Forty-four percent of women with HPV 18 in their tumors had a history of three class I Papanicolaou smears in the 3 years before the diagnosis of cancer, whereas a similar history was elicited in only 16% of those with HPV 16 in their tumors, suggesting that HPV 18-containing tumors might progress to invasion without a prolonged preinvasive period. PMID- 2554229 TI - Migration of simian virus 40 from the vascular system into the glomerular mesangial region. AB - The behavior of simian virus 40 (SV40) injected into the vascular system was investigated in the rat glomerulus. The kidney was perfused via the abdominal aorta with a serum-free culture medium for 5 min, with PBS solution containing SV40 and then with the same medium for 15 min at 37 degrees C. In the glomeruli, SV40 particles were detected in the lumen of the capillaries, fenestrations of endothelial cells and lamina rara interna of the glomerular basement membrane. They were also found in the mesangial matrix and mesangial cells. Invaginations of their membrane were observed on several surface areas where SV40 particles were localized close to the surface. Similarly, when the particles were injected into the tail vein, they were detected in the lamina rara interna, the mesangial matrix, and in vacuoles of mesangial cells at 2 h after the injection. These results indicate that SV40 particles migrate from the vascular system into the mesangial matrix, and are then endocytosed in vivo by mesangial cells. PMID- 2554230 TI - Primary mucinous adenocarcinoma of the eyelid. AB - Primary mucinous adenocarcinoma of the eyelid is a very rare tumor arising from the eccrine sweat gland. It is locally invasive with a high rate of recurrence following removal. Although uncommon, metastasis to regional lymph nodes does occur, and there have been several reports of more distant metastasis. This tumor is characterized by the secretion of mucin. This mucin usually forms into pools surrounding the clusters of tumor cells. Less commonly, the mucin may collect within the tumor cells and not be secreted into the extracellular spaces. Such a "mucin-poor" form of the tumor involving the eyelid was recently seen and is reported here. The clinical and pathologic characteristics of primary mucinous adenocarcinoma are presented and the modes of treatment are discussed. PMID- 2554231 TI - EBV and HSV infections in a patient who had undergone bone marrow transplantation: oral manifestations and diagnosis by in situ nucleic acid hybridization. AB - The course of infections with herpes simplex virus and Epstein-Barr virus in an immunosuppressed patient who had undergone bone marrow transplantation and had tested seronegative for human immunodeficiency virus is described. The clinical oral manifestations were unusual, as they included hairy leukoplakia-like lesions and extensive mucosal ulceration. Histologic examination disclosed unique features consisting of both lichenoid and viral cytopathic changes. The association of the lesions with both Epstein-Barr virus and herpes simplex virus was confirmed by in situ hybridization histochemistry. The importance of recognition of the symptoms, specific diagnosis by DNA hybridization, and implications for antiviral prophylaxis and therapy are emphasized. PMID- 2554232 TI - Characteristics of Epstein-Barr virus-carrying lymphoblastoid cells derived from human tonsil: EBV production and autoantibody response. AB - Tonsillar and peripheral blood lymphocytes from 20 patients with tonsillitis were examined for production of Epstein-Barr virus (EBV) and antinuclear antibodies within 2 weeks after EBV transformation. All established lymphoblastoid cell lines (LCLs) contained more than 90% EBV nuclear antigen-positive cells. However, no LCLs produced biologically active virus. On the other hand, 10 (50%) out of 20 tonsillar LCLs and 4 (20%) out of 20 peripheral blood LCLs produced autoantibodies. These results showed that there are differences in an EBV-induced lymphocyte autoantibody response between tonsillar lymphocytes and peripheral blood lymphocytes. PMID- 2554233 TI - Transitional papilloma of the nasal cavity and paranasal sinuses. Clinical course, viral etiology and malignant transformation. AB - Transitional papilloma is a rare but not uncommon tumor with an overall incidence varying between 0.5 and 4% of all primary nasal tumors. The probability of recurrence is high, between 20 and 62%, even after adequate therapy. Malignancies are associated with transitional papilloma in 2-13% of cases. A series of 21 patients with transitional papilloma of the nose and/or paranasal sinuses is presented. Human papillomavirus (HPV) structural proteins and DNA types were found in 13 cases (62%) of transitional papillomas studied. None of the 9 nasal polyps and none of the 9 squamous cell carcinomas of paranasal sinuses studied as control material contained HPV DNA. We found 4 recurrent papillomas (19%) during the 2-year observation time. All recurrences were HPV-positive. The incidence of malignant transformation was 14% in this series. Treatment of transitional papilloma is always surgical. In our opinion medial maxillectomy and ethmoidectomy via sublabial rhinotomy is the surgery of choice in the treatment of transitional papilloma. PMID- 2554234 TI - A newly isolated avian sarcoma virus, ASV-1, carries the crk oncogene. AB - Avian sarcoma virus 1 (ASV-1) was isolated from a spontaneous sarcoma of an adult chicken. It is a replication-defective virus that induces fibrosarcomas in young chickens and oncogenic transformation in cultured chick embryo fibroblasts. The ASV-1 genome has been cloned in the lambda gt WES.lambda B vector from closed circular viral DNA extracted from infected cells. The nucleotide sequence of the oncogene insert in the ASV-1 genome has been determined. It is virtually identical to the sequence of the oncogene crk recently discovered in the avian sarcoma virus CT10. ASV-1 and CT10 are two independent retrovirus isolates carrying the crk oncogene. PMID- 2554235 TI - The E2 trans-activating protein of bovine papillomavirus type 1 (BPV1) is serine phosphorylated in vivo. AB - The E2 open reading frame of bovine papillomavirus 1 (BPV1) encodes both positive and negative transcriptional regulatory factors. The full-length E2 gene polypeptide is a strong transcriptional transactivator that acts on enhancers within the papillomavirus long control region (LCR), and two shorter E2 proteins function as transcription repressors. A vaccinia recombinant virus harboring the full length E2 coding sequence of BPV1 directs the synthesis of a 48 kD phosphoprotein with specific DNA binding activity. We show that in BPV1 transformed cells the full-length transactivator is a phosphoprotein, whereas truncated E2 proteins were not detectably phosphorylated. PMID- 2554236 TI - Stress-resistance conferred by high level of bcl-2 alpha protein in human B lymphoblastoid cell. AB - High levels of human bcl-2 protein(s) result in (i) the tumorigenic conversion of mouse NIH3T3 cells, (ii) the better survival of mouse myeloid cells in the absence of the required growth factor and (iii) give a growth advantage to human EBV-lymphoblastoid B cells both in low serum medium and limiting dilutions. The effect of the high levels of bcl-2 protein in EBV-B cells was further investigated. This revealed that high levels of bcl-2 alpha protein made EBV-B cells more resistant to a variety of stresses including the application of heat shock, ethanol, methotrexate and the absence of serum. Stress resistance was not observed in EBV-B cells with elevated level of c-myc protein. The mechanism of stress resistance conferred by the bcl-2 alpha protein is yet to be determined although the resistance does not seem to be the result of an increase in major heat shock proteins, hsp70 and hsp90, nor the arrest of cells in G1/G0 phase. The increased viability was observed in control transfectants but not in bcl-2 transfectants when cells are seeded at higher density in the absence of serum. Thus the improved survival of cells as a result of high levels of the bcl-2 alpha protein is not specific to the absence of growth factor but is found to occur with a variety of stresses. PMID- 2554237 TI - Complex interaction of SV40 large T antigen with the control region on the SV40 DNA. AB - SV40 large T antigen interacts specifically with three different DNA sequences within the SV40 control region namely site I, II and III, in order to regulate the viral DNA replication and the stimulation of late, or repression of early viral transcription. We define three different plasmids, containing either site I alone, site II with SV40 specific flanking sequences or both sites I and II with additional neighbouring sequences which were used to study the interaction of T antigen with each of its binding sites. We found that binding of T antigen to site II reduced binding of T antigen to site I as opposed to the interaction of T antigen with site I alone. These results were supported by our findings that T antigen from an SV40 temperature-sensitive mutant, tsA58, which is heat-sensitive for binding to site II, exhibited the same binding affinity for site I as for the DNA fragment containing both binding sites. Thus, binding of T antigen to binding site II seems to interfere with binding to site I, thereby influencing the regulatory functions of T antigen in the repression of early viral gene transcription. PMID- 2554238 TI - Biosynthesis of the protein encoded by the c-met proto-oncogene. AB - The proto-oncogene c-met encodes a transmembrane protein with structural features of a growth factor receptor. We have previously shown that the c-met protein (c Met) is a heterodimer of two disulphide linked chains of 50 kd (alpha) and 145 kd (beta). In this work we have studied the biosynthesis of the c-met product in a gastric carcinoma cell line (GTL-16) where the c-met gene is amplified and overexpressed. Following metabolic labelling of the cells in the presence of tunicamycin, anti-met antibodies immunoprecipitate a protein of 150 kd. In pulse chase experiments carried out in the absence of tunicamycin, a 170 kd product appears first. Within the next few minutes, this precursor modifies its SDS migration, probably as a consequence of modification(s) of its intra-chain disulphide bonds. After 45 min of chase, this single polypeptide precursor is cleaved to form a 50 kd alpha subunit and a 145 kd beta subunit that are joined by disulphide bonds in an alpha beta complex with an apparent molecular weight of 190 kd. The presence of N-linked oligosaccharides in both the precursor and the mature protein was shown by enzymatic de-glycosylation of the immunoprecipitated proteins. The half-life of the mature protein was calculated to be approximately 5h. The c-met protein has similar structure and biosynthesis in other human cell lines. PMID- 2554239 TI - Proto-oncogene amplification and human breast tumor phenotype. AB - Amplification of c-myc, c-erbB-2, hst and int-2 proto-oncogenes was investigated in two independently collected breast tumor series comprising 292 carcinomas. Differences in the frequencies of amplification could be observed between these two series for c-myc (9.3% vs. 20.8%) and hst/int-2 (21.5% vs. 15.6%) whereas similar values were found for c-erbB-2 (22.5% vs. 20.3%). Statistical correlations between amplification and disease parameters were also dependent on population sampling. Therefore we performed our statistical analysis on the pooled populations and focused on the 219 primary breast carcinomas from patients without therapy prior to surgery. Amplification of c-erbB-2 was strongly correlated to the absence of either estrogen (ER-, P = 0.003) or progesterone (PR , P = 0.004) receptors. An amplified c-myc was significantly associated with PR- (P = 0.005) and was prevalent in high grade tumors. On the contrary, hst/int-2 amplification was correlated to PR+ tumors (P = 0.01) and was more frequent in ER+ and low grade tumors, and was also correlated with lymph node involvement (P = 0.04). Our data suggest that amplification of each of these proto-oncogenes could be representative of a particular subset of breast tumors. Therefore, proto oncogene amplification may be helpful in characterizing new biological subclasses in human breast cancer. PMID- 2554240 TI - Transformation of NIH3T3 cells by A-MuLV proviral DNA: effect of plasmid linearization and carrier DNA on transformation efficiency. AB - We have optimized the conditions for efficient NIH3T3 focus formation by calcium phosphate transfection of proviral Abelson-murine leukemia virus (A-MuLV) plasmid DNA. Linearized pA-MuLV, P120 or P160 strains, when transfected with calf thymus carrier DNA, will produce 40-50 foci/100 ng pA-MuLV without co-transfecting Moloney-murine leukemia virus (Mo-MuLV) plasmid DNA. PMID- 2554241 TI - [Influence of morphine and naloxone on the blast transformation of lymphocytes]. AB - The study on the morphine and naloxone influence on the blastic transformation of lymphocytes were made in vitro. The material was taken from the laryngeal cancer and leukoplakia patients. Morphine lowered the threshold of the blastic transformation especially in laryngeal cancer patients. Naloxone on the other hand exaggerated the transformation. This influence may be caused by the lymphocyte opiate receptors. The knowledge of the interdependence between the nervous and immunologic system may be usefull in understanding of the neoplasms pathology mechanisms and constitute a progress especially in oncologic treatment. PMID- 2554242 TI - Recombinant human granulocyte-macrophage colony-stimulating factor primes neonatal granulocytes for enhanced oxidative metabolism and chemotaxis. AB - Recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) induces proliferation and differentiation of hematopoietic stem cells. Additionally, rhGM-CSF enhances the physiologic responses of adult polymorphonuclear leukocytes (PMN) especially with respect to oxidative metabolism and chemotaxis. Neonatal PMN are deficient in chemotaxis and have been demonstrated to have reduced oxidative responses in times of stress. We evaluated the priming effects of rhGM-CSF (1-100 pmol/L) on cord (neonatal) superoxide production and chemotaxis. Cord and adult PMN were incubated with 100 pmol/L rhGM CSF (Amgen, 4 x 10(7) U/mg) for 0-120 min and stimulated with N-formyl-l methionyl-l-leucyl-phenylalanine. RhGM-CSF enhanced O2- production at all time periods with maximal priming at 60 min (147.97 +/- 11.14% p less than or equal to 0.006) with less, but significant enhancement at 120 min (116.53 +/- 7.92% p less than or equal to 0.05). Maximal adult PMN O2- release occurred at 120 min (190.02 +/- 8.71% p less than or equal to 0.003) and was more pronounced than cord PMN.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554243 TI - Adjunctive treatment with low-dose polymyxin B improves operant response rates and latency in infant rats with Haemophilus influenzae type b meningitis. AB - Polymyxin B given in conjunction with ampicillin protects infant rats against death from overwhelming Haemophilus influenzae type b infection. This study was undertaken to examine whether polymyxin B would mitigate the effects of brain damage caused by meningitis. Six- to 7-d-old Sprague-Dawley rats were infected subcutaneously into the nape with 10(7) cfu Haemophilus influenzae type b strain Eagan. This dose consistently caused bacteremia (1.2 X 10(5) cfu/mL) and meningitis (0.5 X 10(5) cfu/mL) in pilot studies. Twenty-four h after infection, all animals received intraperitoneal treatment consisting in either ampicillin alone (400 mg/kg X 4 q 3 h) repeated 12 h later, n = 15 or combined with polymyxin B (0.1 mg/kg/dose) n = 16. At age 2 mo, they were taken off ad libitum feeding and maintained at 80% of their wt. They were then conditioned to receive a food pellet by pressing a lever (continuous reinforcement). The next day, the time lapse between placement and pressing the lever for the first time was recorded (conditioned operant response or latency). Three wk later, the animals were put in the test chamber again and the time to press the lever (latency), as well as the time required to obtain 100 pellets (rate) were recorded. Animals who received polymyxin B had a significantly shorter reaction time; mean 34 s, SEM +/ 5.7 versus mean 88 s, SEM +/- 26.3, P less than or equal to 0.05 and performed significantly faster in obtaining 100 pellets; mean 925 s, SEM +/- 72.1 versus mean 1283 s, SEM +/- 126.3, p less than or equal to 0.02 (analysis of variance, Scheffe test).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554245 TI - [Clinical aspects of viral meningitis in children]. PMID- 2554244 TI - Simultaneous administration of rhesus rotavirus vaccine and oral poliovirus vaccine: immunogenicity and reactogenicity. AB - Rotavirus vaccine could be administered most efficiently if it were incorporated into routine childhood immunizations and did not interfere with the immune response to the other vaccines, principally oral poliovirus vaccine (OPV). We conducted a placebo-controlled randomized trial giving oral rhesus rotavirus vaccine (RRV) (strain MMU 18006) alone and together with a child's first dose of OPV and diphtheria-tetanus toxoids-pertussis to examine the possible interaction of these vaccines. A total of 102 infants 2 to 3 months of age were randomized into 3 groups to receive (1) RRV with OPV, (2) placebo with OPV and (3) RRV 2 weeks after OPV. All infants were given diphtheria-tetanus toxoids-pertussis. Serum samples were collected at the time of OPV immunization and 3 to 5 weeks later. Three to 5 weeks after OPV immunization 60% of infants had a 4-fold rise in neutralization titer to at least one of the three poliovirus serotypes. The rate of antibody response to poliovirus did not differ by RRV groups but a lower rate was correlated with a shorter interval (3 vs. 5 weeks) between OPV vaccination and antibody measurement. Fifty-six percent of infants had a 4-fold rise of IgA and 62% had a 4-fold rise of neutralizing antibody to RRV; this rise did not differ according to time of OPV immunization. RRV was not associated with side effects and may be safely given with OPV to infants 2 to 3 months of age. PMID- 2554246 TI - Kinetic parameters of serum and lung tissue angiotensin-converting enzyme in patients with lung cancer. AB - Angiotensin-converting enzyme activity and its kinetic parameters were determined in both serum and lung tissue samples obtained from 17 patients with lung cancer and 21 control subjects, regardless of ethical base. The specific activities for cancerous and normal lung tissues were 67.2 +/- 27.3 (mean +/- SD) U/g protein and 28.5 +/- 5.4 U/g protein, respectively (P less than 0.001). Serum enzyme activity, on the other hand, was found to be higher in controls (198 +/- 42 U/L) than in patients (237 +/- 68 U/L) (P less than 0.01). The effects of chloride and zinc ions, pH and temperature on the enzyme activity were investigated as kinetic parameters in both serum and tissue samples. PMID- 2554247 TI - Kinetic properties of single sodium channels modified by fenvalerate in mouse neuroblastoma cells. AB - (1) The kinetic properties of single sodium channels modified by the pyrethroid fenvalerate have been analyzed by patch clamp techniques using the cultured mouse neuroblastoma cells. (2) Fenvalerate drastically prolonged the open time of single sodium channels from the normal value of 5 ms to several hundred milliseconds during a depolarizing pulse. The channels remained open after termination of a depolarizing pulse for as long as several seconds. (3) The channel lifetime varied with the membrane potential, attained a maximum at -70 mV, and decreased with hyperpolarization and depolarization from -70 mV. (4) Prolonged openings of the modified channels allowed a current-voltage curve for a single channel to be plotted by sweeping a ramp pulse. The single channel conductance had a value of 11 pS and was linear over potentials ranging from 0 to -100 mV. (5) Power density spectral analysis of the open channel current noise indicated a single Lorentzian curve with a cut-off frequency at 90 Hz, indicating that the increase in noise during channel opening resulted from a relatively slow kinetic process. (6) The probability of the channel being modified by fenvalerate was independent of the length of time during which the channel was opened. This observation suggests that channel modification had taken place before the channel opened. This study of the prolonged opening at the single channel level provides a new insight into open channel properties and the kinetics of channel modification. PMID- 2554248 TI - Characterization of the calcium channel state transitions induced by the enantiomers of the 1,4-dihydropyridine Sandoz 202 791 in neonatal rat heart cells. A nonmodulated receptor model. AB - The actions of the optical enantiomers of Sandoz 202 791 were studied in barium inward currents recorded from single cultured neonatal rat ventricular heart cells, using the whole-cell configuration of the patch clamp technique. The enantiomers were applied by bath perfusion or rapidly by the technique of concentration jumps during single voltage clamp steps. (1) (-)-202 791 reduced the barium current in response to depolarizations positive to 0 mV. The peak current amplitude in the threshold range (-40 to 0 mV) was either not affected or slightly increased by the substance. (2) The agonist enantiomer (+)-202 791 increased the inward current over the whole voltage range, where the increase in peak inward current amplitude was most prominent in the voltage range from -40 mV to 0 mV. (3) The antagonist enantiomer (10(-6) M) induced a 18.2 +/- 2.1 mV (n = 6) shift of the midpoint of the steady state inactivation curve in the hyperpolarizing direction; in contrast (+)-202 791 at the same concentration did cause only a small but not significant shift of the Ca-channel availability curve (n = 5). (4) Rapid extracellular application of (-)-202 791 (10(-6) M), during the sustained current component at a test potential of 0 mV was followed by a sudden acceleration in barium current decay. The drug-induced barium current block developed with a mean time constant of 214.7 +/- 20.6 ms (n = 5). (5) (+) 202 791 (10(-6) M) rapidly applied during test pulses to 0 and -20 mV caused an increase in barium current with a mono- or biexponential time course. The estimated mean time constant of the drug activated Ba2+ current at 0 mV membrane potential was 617.3 +/- 49.3 ms (n = 4). (6) The interaction of Sandoz 202 791 with the Ca-channels is discussed in terms of a "nonmodulated receptor" model. PMID- 2554249 TI - Stimulation of Cl/HCO3 exchange in rat duodenal brush border membrane vesicles by cAMP. AB - Brush border membrane vesicles have been used to study the regulation of rat duodenal HCO3 secretion. When vesicles were loaded with cAMP and ATP SITS sensitive Cl/HCO3 exchange (unidirectional 36Cl influx in response to an outward facing OH/HCO3 gradient) was stimulated by approximately 25%. In contrast, there was no effect of cAMP upon SITS-insensitive 36Cl uptake. The stimulation of Cl/HCO3 exchange caused by cAMP was abolished in the presence of a specific heat stable cAMP-dependent protein kinase inhibitor. These results suggest that Cl/HCO3 exchange, and hence HCO3 secretion, is regulated by cAMP via phosphorylation of some component of the membrane associated with the transport protein. PMID- 2554250 TI - Segmental heterogeneity of basal and aldosterone-induced electrogenic Na transport in human colon. AB - Recent in vitro studies in human colon have demonstrated marked segmental differences in electrogenic Na transport. In the present study, the Na channel blocker amiloride was used further to characterise basal and aldosterone-induced electrogenic Na transport in isolated human distal and proximal colon. Bathed in NaCl Ringer solution, distal and proximal colon exhibited similar basal electrical properties, but the amiloride-sensitive short-circuit current (Isc) was 200% greater in the distal than in the proximal segment. Bathed in choline-Cl Ringer solution, total Isc decreased by 97% in distal colon and by 88% in proximal colon, indicating that Na dependent transport process(es) account almost entirely for the Isc in both segments. Substituting Na2SO4 for NaCl Ringer solution (i) increased amiloride-sensitive Isc by 56% (p less than 0.01) in distal colon but had no effect on amiloride-sensitive Isc in proximal colon, and (ii) decreased amiloride-insensitive Isc in distal and proximal colon by 52% (p less than 0.05) and 81% (p less than 0.001) respectively. After the addition of nystatin to the apical membrane, the relationship between total Isc and mucosal Na concentration indicated that the activity of the basolateral membrane Na pump was similar in both colonic segments. In a further series of experiments, exposure of distal colon to 1 mumol/l aldosterone for 5 h increased total Isc by 52% (p less than 0.05), which reflected stimulation of its amiloride-sensitive component; in contrast, aldosterone had no effect on proximal colon.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554251 TI - Repair synthesis by human cell extracts in DNA damaged by cis- and trans diamminedichloroplatinum(II). AB - DNA damage was induced in closed circular plasmid DNA by treatment with cis- or trans-diamminedichloroplatinum(II). These plasmids were used as substrates in reactions to give quantitative measurements of DNA repair synthesis mediated by cell free extracts from human lymphoid cell lines. Adducts induced by both drugs stimulated repair synthesis in a dose dependent manner by an ATP-requiring process. Measurements by an isopycnic gradient sedimentation method gave an upper limit for the average patch sizes in this in vitro system of around 140 nucleotides. It was estimated that up to 3% of the drug adducts induce the synthesis of a repair patch. The repair synthesis is due to repair of a small fraction of frequent drug adducts, rather than extensive repair of a rare subclass of lesions. Nonspecific DNA synthesis in undamaged plasmids, caused by exonucleolytic degradation and resynthesis, was reduced by repeated purification of intact circular forms. An extract made from cells belonging to xeroderma pigmentosum complementation group A was deficient in repair synthesis in response to the presence of cis- or trans-diamminedichloroplatinum(II) adducts in DNA. PMID- 2554252 TI - Site-specific ribosomal DNA insertion elements in Anopheles gambiae and A. arabiensis: nucleotide sequence of gene-element boundaries. AB - The nucleotide sequence of the junctions between the 28S ribosomal gene and site specific insertion elements from two sibling mosquito species, Anopheles gambiae and A. arabiensis, is reported. In both species, elements insert at the same point within the 28S gene, but this site is 634 basepairs (bp) 3' of the R1 (Type I) insertion site in Drosophila melanogaster. The two mosquito elements each have poly A tails and a polyadenylation signal, but the extreme 3' and 5' ends show no other similarity to each other or to any other insertion element. In both mosquito species, identical target site duplications of 17 bp are generated. The sequence TNTCCCTNT found in this duplication is also found in the 14 bp target site duplications that flank R1 elements in D. melanogaster. Another sequence in this duplication, GGGATAACT, is very similar to the sequence GGGAGTAACT found in the 24 base sequence required by the Bombyx mori R2 endonuclease. PMID- 2554254 TI - A unique sequence of the Bordetella pertussis toxin operon. PMID- 2554253 TI - Magnesium-dependent supercoiling-induced transition in (dG)n.(dC)n stretches and formation of a new G-structure by (dG)n strand. AB - Plasmids containing (dG)27.(dC)27 inserts (pPG27), (dG)37.(dC)37 inserts (pPG37), and (dG)24C(dG)21.(dC)24G(dC)21 inserts (pPG46C) were constructed for the study of structural transitions within (dG)n.(dC)n stretches. Two-dimensional gel electrophoresis has shown that a Mg2+-dependent supercoiling-induced structural transition takes place at pH 8 in plasmid pPG46C. The transition occurs at 0=0.06 and involves a supercoiling release corresponding to 5 superhelical turns. After denaturation of the restriction fragments containing (dG)n.(dC)n inserts, the strands do not renature completely and (dG)n-containing strand migrates in PAGE much faster than the (dC)n-containing one. Chemical modification experiments with the (dG)n-strand have revealed the periodic nature of the protection of guanines against dimethyl sulfate methylation. The (dG)n strand in the presence of Mg2+ forms complexes with the complementary (dC)n strand, which differ from the native duplex in mobility. We believe these effects to be due to the formation of an intrastrand structure within the (dG)n strand stabilized by G.G interactions (we called it G-structure), which in the presence of Mg2+ forms an interstrand complex. with the (dC)n strand. PMID- 2554255 TI - Nucleotide sequence of a rat heart cDNA encoding the isotype alpha of the catalytic subunit of protein phosphatase 2A. PMID- 2554256 TI - Nucleotide sequence of a rat heart cDNA encoding the isotype beta of the catalytic subunit of protein phosphatase 2A. PMID- 2554257 TI - Nucleotide sequence of a cDNA for bovine cytochrome c oxidase subunit VIIc. PMID- 2554258 TI - Sequence of a cDNA from Chlamydomonas reinhardii encoding a ubiquitin 52 amino acid extension protein. PMID- 2554259 TI - Efficient DNA isolation within a single gel barrier tube. PMID- 2554260 TI - [Comparison of blood flow and benzodiazepine receptor distribution in focal epilepsy: preliminary results of a SPECT study]. AB - 99mTc-HMPAO-SPECT and SPECT with the 123I-labelled benzodiazepine (Bz) receptor ligand Ro 16-0154 were performed in 10 patients suffering from partial epilepsy, without cerebral lesion in MRT or CT.2 h p.i. of Ro 16-0154 the distribution of activity correlated with the known distribution of Bz-receptors in the human brain. Perfusion and receptor-binding were found decreased in 7 patients of each study in the suspicious brain-area. 123I-labelled Ro 16-0154 is suitable for Bz receptor mapping by SPECT. The decrease of Bz-receptor binding in epileptic foci, as described in PET-studies, was also detected by SPECT in 7 of 10 patients. PMID- 2554261 TI - [Effect of treatment with enalapril maleate on the levels of circulating catecholamines, beta endorphins, prostaglandins, and concentration of sodium in erythrocytes in patients with essential hypertension]. AB - An effect of enalapril maleate on the activity of renin-angiotensin-aldosterone system and sympathetic reactivity, erythrocyte prostaglandin and sodium levels as well as blood beta-endorphin was investigated in 28 patients with the essential arterial blood hypertension. It was found that enalapril maleate significantly increased plasma renin activity, decreased plasma norepinephrine and its 24-hour excretion, and decreased erythrocyte beta-endorphin and sodium levels. Blood epinephrine and aldosterone levels and their daily excretion remained unchanged similarly to prostaglandins. The above results suggest that a decrease in sympathetic system activity and intracellular sodium concentration may play a role in the hypotensive action of enalapril maleate related to the inhibition of angiotensin II formation. PMID- 2554263 TI - [Activity of angiotensin converting enzyme (ACE) in cushing's disease]. AB - The activity of converting enzyme was determined in 19 patients with Cushing's disease using Friedland and Silverstein technique. Statistically significant increase in the activity of this enzyme was noted in the majority of the examined patients in comparison with healthy subjects. Therefore, inhibitors of the converting enzyme (Captopril) are justified in the treatment of the arterial blood hypertension in Cushing's disease. PMID- 2554262 TI - [Rate of sodium ion outflow through the membranes of lymphocytes and its concentration in the lymphocytes of patients with hyper- and hypothyroidism]. AB - It was shown that sodium ions outflow rate through the lymphocyte membranes is increased in hyperthyroid patients and decreased in patients with hypothyroidism. Sodium ions outflow rate normalizes following effective treatment of the disease. Changes in the sodium ions outflow rate do not effect their concentration in the lymphocytes. PMID- 2554264 TI - Filaggrin expression in cutaneous and mucosal human papillomavirus induced lesions. AB - A series of 32 human papillomavirus induced lesions derived from epidermis and mucosa was studied for the modulation of filaggrin-profilaggrin (F-PF) expression according to the degree of virus infection as compared to normal skin and mucosa biopsies. This investigation was carried out on frozen sections using indirect immunofluorescence for filaggrin detection and group specific viral antigen and by in situ hybridization with biotinylated probes for viral DNA detection and typing. The 9 cutaneous warts showed an increase of F-PF expression in upper layer cells as compared to normal epidermis, which could be related to the high production of virus (viral antigen and HPV types 1 or 2). The 5 condyloma acuminata displayed also an enhanced expression of these components which was located in several upper layers but virus infection was confirmed in 2 of them with HPV types 6, 11 or 16. The 6 laryngeal papillomas exhibited a granular reactivity pattern for F-PF in suprabasal cell layers with an increase in the upper layers; viral antigen was found in 4 cases and HPV DNA types 6, 11 or 16 were detected in 4 specimens. Conversely among 12 cervical intraepithelial neoplasia, F-PF was expressed only in very superficial layers in few cases, without any correlation with the DNA detection (6, 11 or 16, 18). Taken together these data are suggestive of an intense expression of F-PF in benign lesions which can replicate the virus and a discrete or an absent expression of these components in premalignant or malignant lesions. PMID- 2554265 TI - Hepatoid gastric adenocarcinoma. AB - A case of hepatoid gastric adenocarcinoma is reported. The tumor had the histological and immunohistochemical features of both liver cell carcinoma and conventional intestinal-type adenocarcinoma. We discuss the main clinical and pathological features of this uncommon variety of gastric cancer. PMID- 2554266 TI - Mesoblastic nephroma and non-immunological hydrops fetalis. PMID- 2554267 TI - Influence of selection for egg production and Marek's disease resistance on the incidence of endogenous viral genes in White Leghorns. AB - The influence of selection on the frequencies of endogenous viral (ev) genes related to the avian leukosis virus was studied in two genetically distinct sets of White Leghorn strains. Each set consisted of four strains: an unselected control strain, two strains selected for egg production traits, and a strain selected for Marek's disease (MD) resistance as well as egg production traits. Eight different ev genes were observed in Set I and seven in Set II, four being common to both sets. Selection for egg production traits resulted in significant changes of the frequency of four ev genes in both sets. In Set I, increased frequencies were observed for ev-4, ev-7, and ev-8; a decreased frequency for ev 9 was observed. The ev-9 gene expresses the viral envelope protein, whereas the others are transcriptionally silent, with the possible exception of ev-7. In Set II, increased frequencies were observed for the transcriptionally silent ev-8 and for ev-15, a gene which consists of a solitary long terminal repeat. Decreased frequencies were observed for ev-18, which codes for infectious endogenous virus, and for a second ev gene of unknown phenotype. In the resistance-selected strains the frequencies of the ev genes were intermediate between those of the control strains and the strains selected for egg production traits with the exception of ev-6, which expresses the viral envelope protein, and ev-3, which expresses internal viral proteins as well as the envelope protein.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554268 TI - Order of exposure to high dietary calcium and gray strain infectious bronchitis virus alters renal function and the incidence of urolithiasis. AB - Experiments were designed to determine if Gray strain infectious bronchitis virus (IBV) infection increases the incidence of urolithiasis type kidney damage when the urine is already high in Ca and relatively alkaline due to a high Ca-low available P diet (i.e., layer ration). In addition, experiments were conducted to determine the effects of Gray strain IBV on pullet renal function of 6 and 14-wk old pullets at 2, 5, and 10 days postinoculation (PI). Blood gas parameters were measured to determine the mechanism by which layer ration decreases hydrogen ion concentration [( H+]). Urine flow rate, glomerular filtration rate, electrolyte excretion (Na, K, Ca, P), free water clearance, urine osmolality, urine [H+], and renal plasma flow (para-aminohippuric clearance) were measured to assess renal function. Gray strain IBV increased urine [H+] and decreased renal plasma flow in 6-wk-old pullets, and induced a diuresis in 14-wk-old pullets between 5 and 10 days PI. The layer ration increased Ca excretion and induced a metabolic alkalosis, thus decreasing urine [H+] and causing urolith formation. Feeding layer ration followed by Gray strain IBV infection had an additive effect on the incidence of urolithiasis and gross kidney damage. Gray strain IBV infection 8 wk prior to feeding layer ration did not induce urolithiasis. The results suggest that the additive effect of Gray strain IBV on the incidence of urolithiasis is probably due to tubular damage rather than direct changes in renal function parameters. PMID- 2554269 TI - Effect of dietary sodium zeolite A and excess calcium on growth and tibia calcium and phosphorus concentration in uninfected and Eimeria acervulina-infected chicks. AB - Two identical trials were conducted with 5 to 14-day-old broiler chicks. Sodium zeolite A (NZA, 0 and .75%) and Ca (1.0 and 1.5%) were fed to both uninfected and Eimeria acervulina-infected chicks resulting in a 2 x 2 x 2 factorial arrangement of treatments. Coccidial infection reduced weight gain and gain:feed (P less than .01), and tibia ash, Ca, and Ca:P ratio (P less than .05). Excess dietary Ca reduced (P less than .05) weight gain and tibia ash in uninfected chicks but had no effect (P greater than .10) in coccidiosis-infected chicks (Ca x coccidiosis interaction, P less than .05). Addition of NZA to diets with excess Ca further decreased (P less than .05) weight gain and tibia ash in both uninfected and E. acervulina-infected chicks. Tibia Ca, as a percentage of dry fat-free tibia, was reduced (P less than .05) by the addition of NZA. This effect was not observed when tibia Ca was expressed as a percentage of ash. These results suggest that NZA may exacerbate the adverse effects of excess dietary Ca. PMID- 2554270 TI - Evidence for site-specific absorption of a novel ACE inhibitor. AB - Moexipril [2-[(1-ethoxycarbonyl)-3-phenylpropyl]amino-1-oxopropyl]-6, 7-dimethoxy 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (S,S,S)], an ester prodrug of an ACE inhibitor, was formulated in controlled-release preparations with a range of in vitro release rates, to provide a prolonged input of drug in vivo. However, pharmacokinetic studies with the controlled-release dosage forms in humans produced plasma profiles with the same characteristics and time to peak as an immediate-release capsule. In vitro dissolution data from the controlled-release dosage form, as well as the known characteristics of the polymer used to control drug release from the dosage form, suggest no reason to suspect an abrupt halt to the in vivo release of the drug after 1-2 hr. The lack of sustained blood levels is, therefore, most likely due to failure of the GI tract to absorb the drug beyond some location in the upper small intestine, i.e., site-specific absorption. This theory is supported by a series of computer simulations involving moexipril and the active moiety, moexipril diacid. Possible mechanisms include poor drug permeability, a pH effect whereby the zwitterionic form of the drug is more rapidly absorbed, and esterase cleavage of moexipril to the poorly absorbed moexipril diacid. PMID- 2554271 TI - Effects of diethyldithiocarbamate and tetraethylthiuram disulfide on zinc metabolism in mice. AB - Chronic alcoholics with cirrhosis often develop symptoms of zinc deficiency. Tetraethylthiuram disulfide (TTD) is metabolized to two molecules of diethyldithiocarbamate (DDC). DDC chelates divalent metal ions, including zinc, by forming highly lipophilic neutral bis(dithiocarbamate)-metal complexes. DDC could therefore enhance the intestinal zinc uptake or increase the rate of zinc excretion. Accordingly, treatment of alcoholism with TTD could either aggravate or alleviate zinc deficiency. The present study investigated effects of DDC and TTD on intestinal zinc uptake and on the rate of zinc excretion in mice. When given as very high single oral doses, DDC and TTD increased the intestinal uptake of a single oral dose of zinc. When added to the diet and administered in lower doses, closer to those administered to humans for treatment of alcohol abuse, both compounds were without effect on the rate of excretion of the body's zinc stores. In a long-term experiment, where 65Zn was administered in the drinking water, these doses of TTD and DDC reduced the whole-body retention of 65Zn. No treatment changed the organ distribution of zinc in any of the experiments. In conclusion strong indications emerge from the present study that TTD treatment of alcoholism is more likely to reduce the intestinal zinc absorption than to enhance it as has been suggested by other authors. Thus, the widely used experimental model using single oral exposure to metal and chelator conceivably may give erroneous results, when used to predict effects of prolonged exposures. PMID- 2554272 TI - [Primary hyperparathyroidism: value of "intact parathormone" assay (PTH 1-84)]. AB - In a prospective study of 20 patients with primary hyperparathyroidism the diagnostic value of the intact parathormone assay was compared with that of intermediate fragment measurement and that of nephrogenic cyclic AMP determination. In the basal state, measuring the intact parathormone is the best way of separating hyperparathyroid patients form normal subjects. During calcium infusion tests, there is good concordance between changes in intact parathormone and in nephrogenic cyclic AMP. In addition, the intact parathormone assay enables a thorough study of the relationship between ionized calcaemia and parathyroid secretion to be performed and confirms the existence, in hyperparathyroid patients, of two types of secretory response to a rise in calcaemia. Being more reliable than the measurement of a C-terminal or intermediate fragment and easier to perform than nephrogenic cyclic AMP determination, the intact parathormone assay seems to be particularly suitable for the study of primary hyperparathyroidism. PMID- 2554273 TI - [Value of the early diagnosis of cerebral metastasis in non-small cell lung cancers]. PMID- 2554274 TI - [Digestive involvement in AIDS]. AB - The high frequency of gastrointestinal manifestations in AIDS raises new diagnostic and therapeutic problems. In this review paper the authors discuss the various clinical presentations and the different types of macroscopic or histological lesions, as well as the diagnostic approaches. This is followed by a description of the infectious agents responsible for opportunistic or non opportunistic infections in AIDS patients. The epidemiological, clinical and therapeutic aspects of each species are reported. PMID- 2554275 TI - [The effect of mass chest x-ray on early detection of bronchial cancer]. AB - On 30.6.1983, mass radiographic screening was abandoned despite objections to the effect that it offered the only chance of detecting carcinoma of the lung at an early stage. In a retrospective analysis of our patient material comprising 1.010 patients with bronchial carcinomas seen within the period between 1.1.1980 to 31.12.1986, we investigated the question as to whether these objections might not be justified. Four hundred and thirty-seven patients seen prior to 30.6.1983 were compared with 537 patients examined after this date, on the basis of the following parameters: age - sex - symptomatology - cancer stage - histology - treatment. After stopping mass radiographic screening, an increase in age and symptomatic tumour findings was observed, and thus a shift towards more advanced stages of carcinoma. A relative decrease in the number of squamous cell carcinomas, and a relative increase in the number of adenocarcinomas were observed. All the parameters in the patients identified at mass radiographic screening were comparable with those seen in patients identified incidentally. In comparison with symptomatic patients, those identified by mass radiography screening more frequently presented with earlier tumour stages, and the lesions were more frequently resectable. All in all, over a period of three and one-half years, mass radiographic screening revealed 23.8% of all cancers of the lung, and 36.8% of all resectable lesions. After the abandonment of mass radiographic screening, an increase in more advanced and symptomatic tumours was observed. PMID- 2554276 TI - [Biophysical foundations of the effect of cosmic radiation and accelerator radiation]. AB - This book summaries the results of numerous studies, mainly by Soviet investigators, of the characteristics of radiation effects of high energy cosmic rays and of their simulation by heavy charged particle accelerators, and also of their biological effects. It contains the first systematic presentation of data on specific hadron effects. Facts are given showing the ability of these particles to cause profound local damage in tissues and organs. There is evidence that these particles should be considered one of the main factors limiting the duration of space flights. In the book a generalized kinetic model of radiation induced cellular damage also is presented. This model allows an explanations of the observed changes in dose related survival with increasing LET and dose-rate, based on microdosimetric conceptions. PMID- 2554277 TI - [Degradation products of cell receptors and natural antibodies in patients with various forms of pulmonary tuberculosis]. AB - Degradation products of cell receptors, R-proteins, their products complexes with immunoglobulins, hemoreactants and natural antibodies to sheep erythrocytes, were studied in 251 patients with active tuberculosis of the lungs and in 45 healthy persons. The results were indicative of significant changes in the indices as compared to the controls and association of the changes with the clinical form phase and extent of the specific process. PMID- 2554278 TI - Morphologic prognostic factors of malignant mixed mullerian tumor of the uterus: a clinicopathologic study of 58 cases. AB - Sixty-one patients with malignant mixed mullerian tumor (MMMT) of the uterus were seen at L'Hotel-Dieu de Quebec between 1950 and 1986, and the histopathologic material was available for review in 58 cases. Of the morphologic parameters studied, the histologic grade of both epithelial and mesenchymal components correlated with the initial stage. Patients with stages I and II disease had a significantly better survival rate than those with stages III and IV (P less than 0.006). Twenty-five patients with a minimum follow-up of two years were initially stage I or II. All six patients of this subgroup with an epithelial component of serous carcinoma type were dead of their disease at 2 yr. Although not statistically significant, tumor necrosis, the mode of tumor invasive (pushing and infiltrative), and the ratio of epithelial to mesenchymal components tended to influence the outcome. The mitotic rate, the type of mesenchymal components, and vascular and myometrial invasion were of no prognostic significance. We conclude that the outcome of patients with uterine MMMT is mainly influenced by the initial stage and the type of epithelial component. The influence of tumor necrosis, the ratio of epithelial to mesenchymal component, and the mode of tumor invasion deserve further studies. PMID- 2554279 TI - In situ hybridization analysis of human papillomavirus in anal squamous cell carcinoma. AB - To investigate the association between human papillomavirus (HPV) infection and anal carcinoma, we applied a sensitive in situ hybridization technique to detect HPV messenger RNA (HPV m-RNA) in formalin-fixed, paraffin-embedded sections from 18 patients. Using tritium-labeled probes, HPV m-RNA was detected in 12/18 (67%) patients. HPV 6 was detected in four patients, coexisting with HPV 18 in two cases, and HPV 16 was found in eight patients. In six patients, hybridization failed to demonstrate the presence of HPV. With respect to histology, HPV 6 was detected in 1/4 cases of well differentiated invasive squamous cell carcinoma. Ten of thirteen moderately or poorly differentiated invasive squamous cell carcinomas demonstrated HPV m-RNA (HPV 16, eight cases; HPV 6, one case; HPV 6 and 18, one case). HPV 31 was not detected in any specimens. These results suggest that HPV infection may play an important role in the pathogenesis of anal carcinoma. PMID- 2554280 TI - Pulmonary involvement in infectious mononucleosis: histopathologic features and detection of Epstein-Barr virus-related DNA sequences. AB - Symptomatic pulmonary parenchymal involvement is rare in infectious mononucleosis, and few pathologic descriptions of this lesion exist. We describe an unusual patient who presented with adenopathy and rapidly progressive interstitial lung disease. An open lung biopsy showed a predominantly interstitial infiltrate of mononuclear cells distributed along lymphatic routes. A patchy alveolar exudate was also seen. Biopsy of a paratracheal lymph node showed paracortical hyperplasia typical of infectious mononucleosis. The diagnosis was suspected only after thoracotomy and was established by appropriate serological studies for Epstein-Barr virus. Epstein-Barr virus genomic sequences were also detected within routinely processed lung and lymph node tissues using polymerase chain reaction technology. Infectious mononucleosis should be considered in the differential diagnosis of certain lymphoid lung lesions. PMID- 2554281 TI - Malignant fibrous histiocytoma and malignant melanoma: the role of immunohistochemistry and electron microscopy in the differential diagnosis. AB - Malignant fibrous histiocytoma (MFH) and malignant melanoma (MM) occur together often in the differential diagnosis of poorly differentiated neoplasms. They differ, however, in their biologic behavior and recommended treatment. Investigators have therefore explored a variety of special techniques, including electron microscopy (EM) and immunoperoxidase (IP), to classify these tumors accurately and to separate them from each other. To determine the usefulness of IP and EM in the classification of these tumors, we applied a Fontana-Masson stain and IP probes for vimentin, alpha-1-antitrypsin (alpha-1-A), human melanoma black (HMB)-45, and S-100 protein to twelve MMs and nine MFHs, all of which had available EM studies. Three of twelve MMs were amelanotic. All tumors contained vimentin. Eight of twelve MMs and six of nine MFHs contained alpha-1-A. Ten of twelve MMs and no MFHs contained HMB-45. Eleven of twelve MMs and, surprisingly, two of nine tumors classified by light and EM as MFHs contained S-100 protein. When problems arose with either IP negativity or potentially misleading cross reactivity, careful EM study allowed definitive classification. PMID- 2554282 TI - Gastrointestinal stromal tumors: DNA flow-cytometric study of 58 patients with at least five years of follow-up. AB - Flow-cytometrically measured DNA content of 58 surgically resected gastrointestinal stromal tumors (GIST) was correlated with the lesions' histopathologic features and patients' survival. Aneuploid DNA patterns were found in 43 cases (74%), including 33 (76.7%) histologically malignant and 10 (23.3%) in the indeterminate category. DNA ploidy pattern was significantly correlated with mitotic count (less than 5/10 high power field (HPF) versus greater than or equal to 5/10 HPF, P = 0.04) and histologic grade (malignant versus indeterminate, P = 0.02). DNA content was not related to tumor size, site, or histologic patterns (epitheliod versus spindle). The patients' survival was significantly correlated with DNA content (P less than 0.001), histologic grade (indeterminate versus malignant, P = 0.02), and mitotic rate (less than 5/10 HPF versus greater than or equal to 5/10 HPF, P = 0.01). Multivariate regression analysis showed that DNA ploidy is an independent parameter in predicting the clinical outcome for patients with GIST. PMID- 2554283 TI - Primary malignant giant cell tumor of bone: "dedifferentiated" giant cell tumor. AB - Well documented examples of primary malignant giant cell tumor of bone (giant cell tumor and concurrent sarcoma arising de novo) are exceedingly rare in the literature. We report a case arising in the left ischium of a 44-yr-old man. He had no previous history of radiation therapy or multiple resections. Histologically, the tumor was a typical giant cell tumor of bone juxtaposed to a malignant fibrous histiocytoma (MFH). The juxtaposition of a high grade sarcoma (MFH) and a locally aggressive nonmalignant neoplasm such as giant cell tumor is analogous to several other tumors of bone and soft tissue in which a low grade malignant or locally aggressive tumor can be associated with MFH or fibrosarcoma de novo, namely chondrosarcoma, chordoma, liposarcoma, and well differentiated intraosseous and parosteal osteosarcoma. The presence of a high grade malignant component in each of the aforementioned neoplasms generally portends a more ominous prognosis, although this is not invariably true. Recognition of the phenomenon of "dedifferentiation" (or tumor progression) in some bone tumors and sarcomas is important to ensure appropriate treatment. Distinction from secondary malignant giant cell tumors which are usually radiation induced is also important, since the latter have a much worse prognosis than those with dedifferentiation occurring de novo. PMID- 2554284 TI - v-maf, a viral oncogene that encodes a "leucine zipper" motif. AB - We have molecularly cloned the provirus of the avian musculoaponeurotic fibrosarcoma virus AS42. Nucleotide sequence analysis of a biologically active clone of AS42 showed that this virus encodes a viral oncogene, maf. The deduced amino acid sequence of the v-maf gene product contains a "leucine zipper" motif similar to that found in a number of DNA binding proteins, including the gene products of the fos, jun, and myc oncogenes. However, unlike these oncogenes, the cellular maf gene was not transcriptionally activated by growth stimulation of cultured cells. PMID- 2554285 TI - Avian proto-myc genes promoted by defective or nondefective retroviruses are single-hit transforming genes in primary cells. AB - Lymphomas of certain strains of chickens infected by retroviruses frequently contain recombinant transforming genes in which the promoter of the cellular proto-myc gene is replaced by that of a defective rather than an intact retrovirus. Here we ask whether the resulting hybrid genes are sufficient for tumorigenic transformation like viral myc genes. Further, we ask whether retroviruses must be defective in order to mutate proto-myc to a transforming gene or whether the defectiveness plays a transformation-independent function in tumorigenesis. For this purpose the defective provirus of proviral-proto-myc recombinants from lymphomas were repaired, or intact proviruses were recombined with proto-myc genes in vitro, and then compared to recombinant proto-myc genes with defective proviruses for transforming function in quail embryo fibroblasts. It was found that a single copy of a provirus-proto-myc recombinant gene with an intact provirus is sufficient to transform a quail embryo cell in vitro. Moreover, our analyses showed that multiple internal retroviral deletions [corrected] eliminate or inhibit provirus expression. The effect of these deletions [corrected] was detectable only because the inactive proviruses were linked to the selectable, transforming proto-myc gene marker. It is consistent with our results that proviral defectiveness of recombinant proto-myc genes is necessary in vivo for the clonal growth of a transformed cell into a tumor to escape antiviral immunity. The large discrepancy between the probabilities of provirus insertion and tumorigenesis is suggested to reflect the low probabilities of spontaneous deletion of the provirus and of rare, strain specific defects of tumor-resistance genes of the host. PMID- 2554286 TI - Molecular cloning of human testicular angiotensin-converting enzyme: the testis isozyme is identical to the C-terminal half of endothelial angiotensin-converting enzyme. AB - Angiotensin-converting enzyme (ACE; EC 3.4.15.1) is a zinc-containing dipeptidyl carboxypeptidase widely distributed in mammalian tissues and is thought to play a critical role in blood pressure regulation. Testis contains a unique, androgen dependent ACE isozyme of unknown function. We have determined the cDNA sequence for human testicular ACE; it encodes a protein that is identical, from residue 37 to its C terminus, to the second half or C-terminal domain of the endothelial ACE sequence [Soubrier, F., Alhenc-Gelas, F., Hubert, C., Allegrini, J., John, M., Tregear, G. & Corvol, P. (1988) Proc. Natl. Acad. Sci. USA 85, 9386-9390]. The full-length human testis ACE cDNA was constructed from a composite of cloned cDNAs, obtained by a combination of (i) immunoscreening and hybridization screening of a human testicular cDNA library in lambda gt11 and (ii) hybridization screening of human testis cDNAs constructed with ACE-specific primers and amplified by the polymerase chain reaction. The protein sequence inferred consists of a 732-residue preprotein including a 31-residue signal peptide. The mature polypeptide has a molecular weight of 80,073. The testis enzyme contains the second of the two putative metal-binding sites (His-Glu-Met Gly-His) identified in endothelial ACE. This indicates that the functionally active catalytic site is within the C-terminal domain of the endothelial enzyme, accounting for the previous finding that these two structurally dissimilar isozymes are virtually identical catalytically. Of 22 testis ACE cDNAs cloned and sequenced, 3 have unique 5' regions, consisting of inserted, deleted, or substituted sequences up to 328 base pairs long, which have apparently arisen by alternative pre-mRNA splicing. PMID- 2554287 TI - ATP-dependent incorporation of 20S protease into the 26S complex that degrades proteins conjugated to ubiquitin. AB - Previous studies have indicated that the ATP-dependent 26S protease complex that degrades proteins conjugated to ubiquitin is formed by the assembly of three factors in an ATP-requiring process. We now identify one of the factors as the 20S "multicatalytic" protease, a complex of low molecular weight subunits widely distributed in eukaryotic cells. Comparison of the subunit compositions of purified 20S and 26S complexes indicates that the former is an integral part of the latter. By the use of detergent treatment to activate latent protease activity, we show that the 20S protease becomes incorporated into the 26S complex in the ATP-dependent assembly process. It thus seems that the 20S protease is the "catalytic core" of the 26S complex of the ubiquitin proteolytic pathway. PMID- 2554288 TI - Human carboxyl-terminal variant of alpha-type c-erbA inhibits trans-activation by thyroid hormone receptors without binding thyroid hormone. AB - Multiple thyroid hormone (T3; L-3,3',5-triiodothyronine) receptors related to the viral oncogene v-erbA have been identified in mammalian tissues and have been shown to mediate certain actions of T3. The role of a carboxyl-terminal variant of alpha-type c-erbA (c-erbA alpha-2) is controversial, since the human form has been reported to be a T3 receptor, while the rat form has been shown to not bind T3. In fact, the rat homolog of c-erbA alpha-2 has been reported to be an inhibitor of T3 action. We have compared the properties of human c-erbA alpha-2 with those of its rat homolog and with other forms of c-erbA. Neither form of c erbA alpha-2 binds T3 with high affinity whether synthesized in reticulocyte lysates or by transient expression in mammalian cells. Also, neither form increases the expression of a T3-responsive gene. However, human c-erbA alpha-2 inhibits T3 action mediated by a T3-receptor form of c-erbA to an extent similar to that of rat c-erbA alpha-2. Our data strongly suggest that human c-erbA alpha 2 has a biological function similar to that of its rat homolog. Thus, the modulation of T3 action by an endogenous inhibitor related to T3 receptors is likely a general regulatory mechanism. PMID- 2554289 TI - cAMP regulates P450scc gene expression by a cycloheximide-insensitive mechanism in cultured mouse Leydig MA-10 cells. AB - Mouse MA-10 Leydig tumor cells synthesize and secrete progesterone in response to human chorionic gonadotropin, luteinizing hormone, and cAMP but may not synthesize androgens. Maximal doses of human chorionic gonadotropin, ovine luteinizing hormone, forskolin, or 8-bromoadenosine 3',5'-cyclic monophosphate, stimulated cytochrome P450scc mRNA accumulation 1.5- to 3-fold and progesterone secretion 10- to 100-fold in MA-10 cells. P450scc mRNA increased by 2 hr and was maximal by 8 hr; polymerase run-on experiments showed this was due to increased P450scc gene transcription. MA-10 cells are a hormonally homogeneous population, as all cells expressed P450scc mRNA and responded to cAMP equally. cAMP stimulated accumulation of P450scc mRNA continued in the presence of cycloheximide. Gonadotropins stimulated testicular steroidogenesis by coordinate cAMP-induced increases in P450scc gene transcription, mRNA accumulation, and P450scc activity. We cloned rat P450c17 cDNA and showed it detected no P450c17 mRNA in control or cAMP-stimulated MA-10 cells by RNA transfer blots or RNase protection assays. Similarly, HPLC detected no 17 alpha-hydroxyprogesterone or testosterone synthesis in MA-10 cells. Thus MA-10 cells, unlike untransformed Leydig cells, do not express detectable amounts of P450c17 mRNA or P450c17 activity. PMID- 2554290 TI - Hydroxyl radical "footprinting" of RNA: application to pre-mRNA splicing complexes. AB - We present an adaptation of the hydroxyl radical DNA "footprinting" technique that permits high-resolution mapping of protected regions of RNA. Hydroxyl radical cleaves RNA independently of base sequence and secondary structure of the RNAs examined and allows resolution of protected regions at the single nucleotide level. By using this technique, we show that several regions of the 3' splice site of mRNA precursors are protected during the formation of splicing-specific ribonucleoprotein complexes in an in vitro splicing system. These regions include the 3' intron/exon junction and a portion of the adjacent exon, the polypyrimidine tract, and the site of branch formation. These protections appear to be due to splicing specific complexes since their formation is sensitive to point mutations at crucial residues and requires ATP and incubation. The formation of these protected regions is independent of the presence of a 5' splice site. PMID- 2554291 TI - Another version of the human insulin receptor kinase domain: expression, purification, and characterization. AB - We have overexpressed another insulin receptor kinase molecule, which consists of residues 941-1343 inclusive of the human insulin receptor, by using the baculo virus expression vector pVL941. Unlike the two previous preparations of insulin receptor kinase in this expression system, this molecule contains the complete unmodified sequence of the cytoplasmic domain of the human insulin receptor beta subunit. Our construct allows high-level expression of the recombinant protein in cultured Sf9 cells and in cabbage looper (Trichoplusia ni) larvae. An improved purification procedure yields greater than or equal to 95% pure protein in 55% yield from the cells. The specific activity of this purified protein is 3.5-fold greater than from the previously described baculovirus insulin receptor kinase (that included residues 946-1343 from the proreceptor). Like the latter molecule, the insulin receptor kinase molecule reported here sediments as a monomer, and its autophosphorylation occurs by an intramolecular process. Preliminary data about the spectroscopic features of the cytosolic domain of the human insulin receptor are presented. PMID- 2554292 TI - Increases in Na+,K+-ATPase activity of erythrocytes and skeletal muscle after chronic ethanol consumption: evidence for reduced efficiency of the enzyme. AB - Increased Na+,K+-ATPase activity observed after chronic ethanol consumption has been examined to determine whether the increase is due to changes in the kinetic properties of the enzyme or increases in the amount of enzyme in the membranes examined. In skeletal muscle and erythrocyte ghosts from rat, as well as from humans, increased Na+,K+-ATPase activity in ethanol-consuming individuals was not accompanied by an increase in the number of ouabain binding sites. In studies with intact human erythrocytes, similar ouabain-sensitive 22Na+ and 86Rb+ pumping rates were observed between normal and ethanol-consuming individuals and the Na+ to Rb+ pumping ratio was found to be 1.5 in all cases. However, ouabain-sensitive lactate plus Pi formation was increased in cells from alcoholic individuals. Thus these data suggest that increased enzyme activity may be due to a kinetic alteration of the Na+,K+-ATPase and that the enzyme may be less efficient in coupling ion pumping to ATP hydrolysis than the enzyme in normal cells. PMID- 2554293 TI - Structural changes in bacteriorhodopsin during proton translocation revealed by neutron diffraction. AB - A neutron diffraction study of spectroscopic states for the light-energized proton pump bacteriorhodopsin (BR) is presented. The photocycle states BR-568 and M were generated at temperatures above 4 degrees C and were measured after trapping at--180 degrees C. In the BR-568 to M-state transition, which is known to be a key step in transmembrane proton pumping, reversible structural changes of the protein were detected. These structural alterations occur in the neighborhood of the cyclohexene ring and at the Schiff's base end of the chromophore retinal. They are interpreted as a 1-2 degree tilt of three or four of the transmembrane alpha-helices or as positional changes of four or five amino acids. The structural changes observed are inherent in the transport mechanism of bacteriorhodopsin. PMID- 2554294 TI - Stimulation of beta-adrenergic receptors of S49 lymphoma cells redistributes the alpha subunit of the stimulatory G protein between cytosol and membranes. AB - The stimulatory guanine nucleotide-binding protein (Gs), which links cell-surface receptors to second-messenger effector systems, is assumed to be confined to plasma membranes. In the current studies we tested whether Gs redistributes within cells by treating S49 lymphoma cells with the beta-adrenergic agonist isoproterenol, then separating cytosol and crude membrane fractions (defined as pellet and supernatant, respectively, after centrifugation for 1 hr at 150,000 x g), and assaying fractions for the alpha subunit of Gs (alpha s) using a competitive ELISA and reconstitution techniques. Under basal conditions, a small (10%) pool of alpha s was identified in supernatant fractions of S49 cells. The size of this pool decreased in the first 15 min after agonist treatment of cells. This decrease was blocked by a beta-adrenergic receptor antagonist and did not occur in an S49 variant, UNC, which lacks functional interaction between receptors and Gs. The size of the alpha s pool in supernatant fractions increased to almost 50% of total cellular alpha s during a 1-hr incubation of cells with isoproterenol. Before isoproterenol treatment only the competitive ELISA was sensitive enough to detect cytosolic alpha s, whereas at later time points (greater than or equal to 30 min) the presence of alpha s in the cytosol was confirmed by both immunoblotting and by reconstitution of adenylyl cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] in Gs-deficient membranes derived from cyc-S49 cells. In contrast to membrane alpha s, cytosolic alpha s did not require activation (e.g., by AlF4-) in the reconstitution assay to stimulate adenylyl cyclase. Use of an antibody that selectively recognizes monomeric dissociated alpha s, but not heterotrimeric alpha s, indicated that cytosolic alpha s is monomeric. These data indicate that alpha s is not exclusively localized to the plasma membrane and that agonist treatment redistributes this protein within target cells. PMID- 2554295 TI - Vertebrate and yeast calmodulin, despite significant sequence divergence, are functionally interchangeable. AB - Yeast strains relying solely on vertebrate (Xenopus laevis) calmodulin, expressed under control of a yeast (GAL1) promoter, grew at the same rate as yeast cells containing their endogenous calmodulin. Therefore, the ability to perform essential functions has been conserved between yeast and vertebrate calmodulins, suggesting that calmodulin performs the same (or overlapping) roles in yeast as it does in higher eukaryotes. Successful substitution of vertebrate for yeast calmodulin also suggests that the two proteins can adopt similar conformations in vivo, despite the large number of amino acid differences between them (60 out of 148 residues). Strains overproducing either vertebrate or yeast calmodulin about 100-fold and a strain producing a normal level of yeast calmodulin were essentially indistinguishable in many characteristics, including microtubule distribution, rate of secretion, response to mating pheromone, sporulation, and adaptation to nutrient limitation. Calmodulin overproduction did not confer elevated resistance to a phenothiazine drug, trifluoperazine, thought to be a calmodulin-specific inhibitor. These results have important implications for understanding the role of calmodulin in intracellular calcium signaling. PMID- 2554296 TI - Mouse NIH 3T3 cells expressing human colony-stimulating factor 1 (CSF-1) receptors overgrow in serum-free medium containing human CSF-1 as their only growth factor. AB - Mouse NIH 3T3 cells expressing the human c-fms protooncogene encoding the receptor for colony-stimulating factor 1 (CSF-1) are able to proliferate in serum free medium containing platelet-derived growth factor (PDGF), insulin, transferrin, and albumin as the only exogenous proteins. When PDGF and insulin were replaced by purified human recombinant CSF-1, the cells became spindle shaped and refractile, were no longer contact inhibited, and proliferated to high densities. Thus, transduction of the human c-fms gene into mouse fibroblasts can not only reprogram their growth factor requirements but can also induce ligand dependent features of cell transformation. NIH 3T3 cells stably transformed by the feline v-fms oncogene or by a mutated, oncogenic human c-fms gene were able to proliferate in the absence of exogenous growth factors. A monoclonal antibody that prevents signal transduction by the human CSF-1 receptor inhibited the growth of cells transformed by the activated c-fms oncogene, confirming that CSF 1 receptor function was required to abrogate growth factor requirements and to maintain the transformed state. PMID- 2554297 TI - Spontaneous Kearns-Sayre/chronic external ophthalmoplegia plus syndrome associated with a mitochondrial DNA deletion: a slip-replication model and metabolic therapy. AB - The muscle mitochondria of a patient with Kearns-Sayre/chronic external ophthalmoplegia plus syndrome were found to be completely deficient in respiratory complex I activity and partially deficient in complex IV and V activities. Treatment of the patient with coenzyme Q10 and succinate resulted in clinical improvement of respiratory function, consistent with the respiratory deficiencies. Restriction enzyme analysis of the muscle mtDNA revealed a 4.9 kilobase deletion in 50% of the mtDNA molecules. Polymerase chain reaction analysis demonstrated that the deletion was present in the patient's muscle but not in her lymphocytes or platelets. Furthermore, the deletion was not present in the muscle or platelets of two sisters. Hence, the mutation probably occurred in the patient's somatic cells. Direct sequencing of polymerase chain reaction amplified DNA revealed a 4977-base-pair deletion removing four genes for subunits of complex I, one gene for complex IV, two genes for complex V, and five genes for tRNAs, which paralleled the respiratory enzymes affected in the disease. A 13 base-pair direct repeat was observed upstream from both breakpoints. Relative to the direction of heavy-strand replication, the first repeat was retained and the second repeat was deleted, suggesting a slip-replication mechanism. Sequence analysis of the human mtDNA revealed many direct repeats of 10 base pairs or greater, indicating that this mechanism could account for other reported deletions. We postulate that the prevalence of direct repeats in the mtDNA is a consequence of the guanine-cytosine bias of the heavy and light strands. PMID- 2554298 TI - Transmembrane domain of the envelope gene of a polycythemia-inducing retrovirus determines erythropoietin-independent growth. AB - Both the polycythemia-inducing and the anemia-inducing strains of Friend spleen focus-forming virus can induce acute erythroleukemia in susceptible adult mice. However, only cells infected with the polycythemia-inducing strain become erythropoietin-independent for proliferation and differentiation. The sequences responsible for the altered erythropoietin responsiveness have previously been localized to a 678-base-pair EcoRI-Cla I fragment at the 3' end of the envelope gene. This region is now further analyzed by dividing it into two fragments by using the Fok I restriction site. Two recombinants were made by replacing either the 558-base-pair EcoRI-Fok I or the 113-base-pair Fok I-Cla I env gene fragments from the anemia-inducing strain of spleen focus-forming virus with sequences derived from the polycythemia-inducing strain. Our results indicate that the 113 base-pair Fok I-Cla I fragment, which encodes primarily the transmembrane domain of the envelope protein, determines erythropoietin-independent growth. PMID- 2554299 TI - alar4, a constitutive mutant of the A system for amino acid transport, has increased abundance of the Na+,K+-ATPase and mRNA for alpha 1 subunit of this enzyme. AB - A constitutive mutant, alar4, for the A system of amino acid transport, has increased activity and amount of the A system. This is accompanied by increased sensitivity to ouabain, as measured by efficiency of plating, and increased activity and abundance of the Na+,K+-ATPase that is present in the parental cell line, CHO-K1 (wild type). The latter was shown by increases in (i) ouabain inhibitable 86Rb uptake in intact cells, (ii) ouabain-inhibitable ATPase activity in mixed membrane vesicles, and (iii) number of ouabain-binding sites and by similar Kd values for ouabain binding and K1/2 for ouabain inhibition of Na+,K+ ATPase as compared to the wild type. The increase in abundance of the Na+ pump is associated with a 4-fold increase in abundance of the mRNA for the alpha 1 subunit of the Na+,K+-ATPase. We could not detect mRNA for alpha 2 or alpha 3 or for the beta subunits. The increase in abundance of the A system and Na+,K+ ATPase is associated with a negligible increase in intracellular Na+ concentration. We propose that the increase in the abundance of the A system and the Na+,K+-ATPase is the result of a mutation in regulatory gene R1 that controls the A system and the Na+,K+-ATPase and is not due to a primary effect of a possible initial increase in Na+ concentration. PMID- 2554300 TI - Construction of a recombinant human parvovirus B19: adeno-associated virus 2 (AAV) DNA inverted terminal repeats are functional in an AAV-B19 hybrid virus. AB - To facilitate genetic analysis of the human pathogenic parvovirus B19, we constructed a hybrid B19 viral genome in which the defective B19 inverted terminal repeats were replaced with the full-length inverted terminal repeats from a nonpathogenic human parvovirus, the adeno-associated virus 2 (AAV). The hybrid AAV-B19 genome was rescued from a recombinant plasmid and then the DNA was replicated upon transfection into adenovirus 2-infected human KB cells in the presence of AAV genes coding for proteins required for AAV DNA replication (AAV Rep proteins). In addition, in the presence of AAV genes coding for the viral capsid proteins (AAV-Cap proteins), the rescued/replicated hybrid AAV-B19 genomes were packed into mature AAV progeny virions, which were subsequently released into culture supernatants. The recombinant AAV-B19 progeny virions were infectious for normal human bone marrow cells and strongly suppressed erythropoiesis in vitro. The availability of an infectious recombinant B19 virus should facilitate the mutational analysis of the viral genome, which, in turn, may yield information on individual viral gene functions in B19-induced pathogenesis. The hybrid AAV-B19 genome may also prove to be a useful vector for gene transfer in human bone marrow cells. PMID- 2554301 TI - Inhibition of inactivation of single sodium channels by a site-directed antibody. AB - The effects of site-directed antibodies on single sodium channel currents in excised membrane patches from rat brain neurons have been examined. Of six antibodies directed against different intracellular domains of the sodium channel alpha subunit, only an antibody directed against a highly conserved intracellular segment between homologous transmembrane domains III and IV induced late single channel openings and prolonged single channel open times during depolarizing test pulses, resulting in nearly complete inhibition of sodium channel inactivation. The antibody effect was not observed if the membrane patches were depolarized to inactivate sodium channels before exposure to the antibody, indicating that the intracellular sequence recognized by the antibody is rendered inaccessible by inactivation. The results show that a conformational change involving the intracellular segment between domains III and IV of the alpha subunit of the sodium channel molecule is required for fast sodium channel inactivation and suggest that this segment may be the fast inactivation gate of the sodium channel. PMID- 2554302 TI - Molecular cloning of a putative tetrodotoxin-resistant rat heart Na+ channel isoform. AB - Voltage-gated Na+ channels in mammalian heart differ from those in nerve and skeletal muscle. One major difference is that tetrodotoxin (TTX)-resistant cardiac Na+ channels are blocked by 1-10 microM TTX, whereas TTX-sensitive nerve Na+ channels are blocked by nanomolar TTX concentrations. We constructed a cDNA library from 6-day-old rat hearts, where only low-affinity [3H]saxitoxin receptors, corresponding to TTX-resistant Na+ channels, were detected. We isolated several overlapping cDNA clones encompassing 7542 nucleotides and encoding the entire alpha subunit of a cardiac-specific Na+ channel isoform (designated rat heart I) as well as several rat brain I Na+ channel cDNA clones. The derived amino acid sequence of rat heart I was highly homologous to, but distinct from, previous Na+ channel clones. RNase protection studies showed that the corresponding mRNA species is abundant in newborn and adult rat hearts, but not detectable in brain or innervated skeletal muscle. The same mRNA species appears upon denervation of skeletal muscle, likely accounting for expression of new TTX-resistant Na+ channels. Thus, this cardiac-specific Na+ channel clone appears to encode a distinct TTX-resistant isoform and is another member of the mammalian Na+ channel multigene family, found in newborn heart and denervated skeletal muscles. PMID- 2554303 TI - The mRNA encoding a high-affinity cAMP phosphodiesterase is regulated by hormones and cAMP. AB - To elucidate the mechanisms by which hormones regulate cAMP phosphodiesterases (PDEs), a group of cDNA clones that had been isolated from a rat Sertoli cell library were characterized. These cDNAs are derived from a single gene (ratPDE3). The deduced amino acid sequence of the ratPDE3 cDNA corresponds to a 66,200-Da protein homologous to other testicular PDEs, to the Drosophila melanogaster dunce encoded cAMP PDE, and to bovine and yeast PDEs. Expression of ratPDE3 in eukaryotic and prokaryotic cells leads to the appearance of a cAMP PDE with properties identical to the cAMP PDE purified from Sertoli cells. Although of different size, transcripts corresponding to ratPDE3 were present in all organs studied. In the immature Sertoli cell in culture, the level of mRNA transcripts of ratPDE3 was increased more than 100-fold by follicle-stimulating hormone or N6,O2'-dibutyryladenosine 3',5'-cyclic monophosphate treatment. Stimulation of ratPDE3 mRNA by N6,O2'-dibutyryladenosine 3',5'-cyclic monophosphate was also observed in a C6 glioma cell line. These data demonstrate that cAMP regulates the expression of one of its own degrading enzymes by an intracellular feedback mechanism that involves changes in mRNA levels. PMID- 2554304 TI - Human 72-kilodalton type IV collagenase forms a complex with a tissue inhibitor of metalloproteases designated TIMP-2. AB - Simian virus 40 (SV40)-transformed human lung fibroblasts secrete both 72-kDa type IV collagenase and a closely related 92-kDa type IV collagenase that was not detected in the parental cell line. The 92-kDa type IV procollagenase purified from these cells exists in a noncovalent complex with the tissue inhibitor of metalloproteases, TIMP. Here we report that the 72-kDa type IV procollagenase purified from HRAS-transformed human bronchial epithelial cells, SV40-transformed lung fibroblasts, and normal skin fibroblasts exists in a stable but noncovalent stoichiometric complex with a 24-kDa inhibitor referred to here as "TIMP-2." TIMP 2 is closely related to TIMP, as demonstrated by comparison of the partial amino acid sequence of this protein to that of TIMP, although it does not cross-react with TIMP-specific antibody. The TIMP-2 inhibitor interacts with the 72-kDa type IV collagenase in preference to the 92-kDa type IV collagenase that forms a complex exclusively with TIMP. The 72-kDa type IV collagenase-TIMP-2 complex can be activated with organomercurials to yield a catalytically competent enzyme. Activation occurs concomitantly with autoproteolytic cleavage of the amino terminus of the protein and does not require dissociation of the complex. Both activity and activation of the complex can be completely inhibited by further addition of stoichiometric quantities of purified TIMP-2 or recombinant TIMP. PMID- 2554305 TI - Platelet-derived growth factor (PDGF) binding promotes physical association of PDGF receptor with phospholipase C. AB - Phospholipase C (PLC)-mediated production of inositol trisphosphate and diacylglycerol is stimulated by binding of platelet-derived growth factor (PDGF) to cell-surface receptors. Antibodies recognizing native PDGF receptors through peptide-domain epitopes coprecipitated 4-fold more PLC activity with receptors from PDGF-stimulated than from unstimulated BALB/c 3T3 cells, despite equivalent PDGF receptor recovery. Activity coprecipitated from unstimulated cells was comparable to nonspecific activity recovered with preimmune sera or in the presence of competing peptide immunogen. PLC activity coprecipitating with PDGF receptors represented 60% of anti-phosphotyrosine antibody-recovered activity from PDGF-stimulated cells. Coprecipitation was rapidly induced in cells treated with PDGF at 4 degrees C, reversibly lost with acid dissociation of PDGF from receptors, and recovered with PDGF readdition. PDGF concentrations effecting coprecipitation correlated with stimulation of intact-cell inositol phosphate production. Monoclonal antibodies to PLC gamma (145 kDa) coprecipitated from PDGF stimulated cell lysates (but not from unstimulated cell lysates) tyrosine kinase activity that phosphorylated PDGF receptor and PLC gamma. Stable physical association of PDGF receptors with PLC may participate in coupling ligand binding to increased PLC activity. PMID- 2554306 TI - In vivo synthesis of adenylylated bis(5'-nucleosidyl) tetraphosphates (Ap4N) by Escherichia coli aminoacyl-tRNA synthetases. AB - The role of aminoacyl-tRNA synthetases in the in vivo synthesis of adenylylated bis(5'-nucleosidyl) tetraphosphates (Ap4N) was studied by measuring the concentration of these nucleotides in Escherichia coli cells overproducing lysyl , methionyl- phenylalanyl-, or valyl-tRNA synthetase. Overproduction of each aminoacyl-tRNA synthetase (20- to 80-fold) was accompanied by a significant increase in intracellular Ap4N concentration (3- to 14-fold). As expected, non adenylylated bis(5'-nucleosidyl) tetraphosphate concentration was not changed by synthetase overproduction. It was also verified that overproduction of an inactive methionyl-tRNA synthetase mutant did not modify Ap4N concentration. Ap4N accumulation during heat shock occurred in all strains studied. The increase factor (approximately 50-fold after 1 hr at 48 degrees C) was not changed by overproduction of any of the aminoacyl-tRNA synthetases studied, including that of the heat-inducible form of lysyl-tRNA synthetase from the lysU gene. Together, these results establish that aminoacyl-tRNA synthetases are involved in Ap4N biosynthesis during exponential growth as well as during heat shock. PMID- 2554307 TI - H-2RIIBP, a member of the nuclear hormone receptor superfamily that binds to both the regulatory element of major histocompatibility class I genes and the estrogen response element. AB - Transcription of major histocompatibility complex (MHC) class I genes is regulated by the conserved MHC class I regulatory element (CRE). The CRE has two factor-binding sites, region I and region II, both of which elicit enhancer function. By screening a mouse lambda gt 11 library with the CRE as a probe, we isolated a cDNA clone that encodes a protein capable of binding to region II of the CRE. This protein, H-2RIIBP (H-2 region II binding protein), bound to the native region II sequence, but not to other MHC cis-acting sequences or to mutant region II sequences, similar to the naturally occurring region II factor in mouse cells. The deduced amino acid sequence of H-2RIIBP revealed two putative zinc fingers homologous to the DNA-binding domain of steroid/thyroid hormone receptors. Although sequence similarity in other regions was minimal, H-2RIIBP has apparent modular domains characteristic of the nuclear hormone receptors. Further analyses showed that both H-2RIIBP and the natural region II factor bind to the estrogen response element (ERE) of the vitellogenin A2 gene. The ERE is composed of a palindrome, and half of this palindrome resembles the region II binding site of the MHC CRE. These results indicate that H-2RIIBP (i) is a member of the superfamily of nuclear hormone receptors and (ii) may regulate not only MHC class I genes but also genes containing the ERE and related sequences. Sequences homologous to the H-2RIIBP gene are widely conserved in the animal kingdom. H-2RIIBP mRNA is expressed in many mouse tissues, in agreement with the distribution of the natural region II factor. PMID- 2554308 TI - Cell proteins bind to multiple sites within the 5' untranslated region of poliovirus RNA. AB - The 5' noncoding region of poliovirus RNA contains sequences necessary for translation and replication. These functions are probably carried out by recognition of poliovirus RNA by cellular and/or viral proteins. Using a mobility shift electrophoresis assay and 1,10-phenanthroline/Cu+ footprinting, we demonstrate specific binding of cytoplasmic factors with a sequence from nucleotides 510-629 within the 5' untranslated region (UTR). Complex formation was also observed with a second sequence (nucleotides 97-182) within the 5' UTR. These two regions of the 5' UTR appear to be recognized by distinct cell factors as determined by competition analysis and the effects of ionic strength on complex formation. However, both complexes contain eukaryotic initiation factor 2 alpha, as revealed by their reaction with specific antibody. PMID- 2554309 TI - Independent expression of human alpha or beta platelet-derived growth factor receptor cDNAs in a naive hematopoietic cell leads to functional coupling with mitogenic and chemotactic signaling pathways. AB - Distinct genes encode alpha and beta platelet-derived growth factor (PDGF) receptors that differ in their abilities to be triggered by three dimeric forms of the PDGF molecule. We show that PDGF-receptor mitogenic function can be reconstituted in a naive hematopoietic cell line by introduction of expression vectors for either alpha or beta PDGF receptor cDNAs. Thus, each receptor is independently capable of coupling with mitogenic signal-transduction pathways inherently present in these cells. Activation of either receptor also resulted in chemotaxis, alterations in inositol lipid metabolism, and mobilization of intracellular Ca2+. The magnitude of these functional responses correlated well with the binding properties of the different PDGF isoforms to each receptor. Thus, availability of specific PDGF isoforms and relative expression of each PDGF receptor gene product are major determinants of the spectrum of known PDGF responses. PMID- 2554310 TI - Transcription terminates near the poly(A) site in the CYC1 gene of the yeast Saccharomyces cerevisiae. AB - A 38-base-pair region required for normal CYC1 mRNA 3' end formation in Saccharomyces cerevisiae was shown to be necessary for the termination of transcription in vivo by examining the stability of CEN3 plasmids. CEN3 plasmids were stably maintained during vegetative growth, unless a GAL1 transcript impinged on the CEN3 region. Transcription from the GAL1 promoter was terminated, and plasmid stability was restored by the insertion of a fragment containing the 38-base-pair region of CYC1. In contrast, a similar fragment lacking the 38-base pair region had no such stabilizing effect. Furthermore, CYC1 mRNA transcription terminated in a region less than 100 nucleotides downstream from the normal poly(A) site, thus establishing that CYC1 mRNA 3' end formation does not involve overly extended precursors as are observed in higher eukaryotes. PMID- 2554311 TI - Pertussis holotoxoid formed in vitro with a genetically deactivated S1 subunit. AB - The cytotoxicity of pertussis toxin, a multisubunit exotoxin produced by Bordetella pertussis, is believed to be due to the ADP-ribosyltransferase activity of the S1 subunit. We have previously described the recombinant expression of each of the five individual pertussis toxin subunits in Escherichia coli and the production of an enzymatically deficient form of the S1 subunit by site-directed mutagenesis. We now report the in vitro assembly of holotoxin from native pertussis toxin B oligomer and recombinant S1 subunits, the latter purified and refolded from insoluble inclusion bodies. Holotoxin assembled with recombinant S1 of authentic amino acid sequence was indistinguishable from native pertussis toxin in its electrophoretic migration and ability to elicit a cytopathic response in cultured Chinese hamster ovary cells; in contrast, holotoxin assembled with the genetically deactivated analog of recombinant S1 displayed greatly diminished cytopathicity. These results verify that the in vitro cytopathic effects of pertussis toxin are the result of the enzymatic activity of the S1 subunit and illustrate the potential for constructing complex quaternary protein structures in vitro from insoluble, unfolded polypeptides derived from expression in recombinant systems. PMID- 2554312 TI - "Activated"-RecA protein affinity chromatography of LexA repressor and other SOS regulated proteins. AB - We have developed an affinity column to study the interaction of LexA repressor and other substrates with the activated form of RecA protein. Nucleoprotein complexes of RecA protein, (dT)25-30, and adenosine 5'-[gamma-S]thio-triphosphate were formed in solution and bound to RecA protein-agarose columns. These "activated"-RecA nucleoprotein complexes were retained by strong hydrophobic interactions. Purified LexA protein bound tightly to these activated RecA columns, whereas the LexA protein bound poorly to RecA-agarose alone. Once bound, LexA protein underwent specific proteolysis, and the fragments were released from the complex. The mutant LexA protein, LexA-SA119, which cannot carry out self cleavage or RecA-mediated cleavage in solution, bound efficiently to the activated RecA column but was not cleaved, indicating that these columns can be used to identify residues involved in RecA-LexA binding. As an example of this use, nucleoprotein complexes were prepared using the RecA430 protein. In vivo the recA430 mutation blocks induction of the SOS response. LexA protein was not efficiently retained on the immobilized RecA430 complexes, suggesting that Gly 204 is required for efficient repressor binding. These results show that activated RecA affinity columns can be used to investigate the binding and cleaving properties of mutationally altered RecA and LexA proteins. Additionally, these activated RecA columns have been used to investigate binding interactions of phage lambda repressor, as well as the UmuC protein, which is required for chemical mutagenesis. PMID- 2554313 TI - Isoproterenol stimulates phosphorylation of the insulin-regulatable glucose transporter in rat adipocytes. AB - We have examined the acute effects of insulin and isoproterenol on the phosphorylation state of the insulin-regulatable glucose transporter (IRGT) in rat adipocytes. The IRGT was immunoprecipitated from either detergent-solubilized whole-cell homogenates or subcellular fractions of 32P-labeled fat cells and subjected to sodium dodecyl sulfate/polyacrylamide gel electrophoresis. The 32P labeled IRGT was detected by autoradiography as a species of apparent Mr 46,000. Insulin stimulated translocation of the IRGT from low-density microsomes to the plasma membrane but did not affect phosphorylation of the transporter in either fraction. Isoproterenol inhibited insulin-stimulated glucose transport by 40% but was without effect on the subcellular distribution of the transporter in either the presence or absence of insulin. Isoproterenol stimulated phosphorylation of the IRGT 2-fold. Incubating cells with dibutyryl-cAMP and 8-bromo-cAMP also stimulated phosphorylation 2-fold, and the transporter was phosphorylated in vitro when IRGT-enriched vesicles were incubated with cAMP-dependent protein kinase and [gamma-32P]ATP. These results suggest that isoproterenol stimulates phosphorylation of the IRGT via a cAMP-dependent pathway and that phosphorylation of the transporter may modulate its ability to transport glucose. PMID- 2554314 TI - Direct observations of ligand dynamics in hemoglobin by subpicosecond infrared spectroscopy. AB - The photodissociation of CO from HbCO at ambient temperature is studied by means of a femtosecond IR technique. The bleaching of the FeCO absorption and the appearance of a new IR absorption near that of free CO are both observed at 300 fs after optical excitation. The bleach does not recover on the time scale of a few picoseconds but does recover by approximately 4% within 1 ns, which suggests that a barrier to recombination is formed within a few picoseconds. The CO spectrum does not change significantly between 300 fs and 1 ns, suggesting that the CO quickly finds some locations in the heme pocket that are not more than a few angstroms from the iron. The de-ligated CO appears in its ground vibrational level. There is evidence that 85 +/- 10% of this CO remains in the heme pocket at 1 ns; it probably resides there for 50 ns. The flow of excess vibrational energy from the heme to the solvent was directly observed in the IR experiments. The heme cools within 1-2 ps while thermal disruption of the surrounding solvent structure requires approximately 30 ps. PMID- 2554315 TI - Detection and mapping of spliced RNA from a human hepatoma cell line transfected with the hepatitis B virus genome. AB - HepG2 cells, known to support the replication and virion formation of hepatitis B virus (HBV), were transfected with a cosmid constructed to contain 12 tandem head to-tail repeats of the HBV genome for effective HBV genome expression. We detected previously identified RNAs of 3.3, 2.3, and 2.0 kilobases (kb) that code for core antigen, large surface antigen, and middle/major surface antigen, respectively. We also detected four additional RNAs of 2.1, 1.7, 1.1, and 0.7 kb [the lengths exclude the poly(A) tail]. S1 mapping and nucleotide sequencing data showed that the 2.1-kb RNA is a spliced RNA whose 5' and 3' ends are identical to those of the 3.3-kb RNA. The results suggest that the 2.1-kb RNA codes for an altered core antigen lacking the last amino acid, cysteine, and that expression of the 3.3-kb pregenomic RNA is regulated, at least in part, by splicing. The map positions of the 1.7- and 1.1-kb RNAs suggest that they code for the carboxyl terminal portions of the putative polymerase, whereas the 0.7-kb RNA codes for the X protein. PMID- 2554316 TI - Translocation of proteins across the mitochondrial inner membrane, but not into the outer membrane, requires nucleoside triphosphates in the matrix. AB - Work in several laboratories has established that import of proteins across the mitochondrial inner membrane requires an electrochemical potential across that membrane and cleavage of nucleoside triphosphate. We now show that nucleoside triphosphate must be present inside the inner membrane. In contrast, the potential-independent insertion of porin into the outer membrane requires ATP only outside the inner membrane. PMID- 2554317 TI - AbrB, a regulator of gene expression in Bacillus, interacts with the transcription initiation regions of a sporulation gene and an antibiotic biosynthesis gene. AB - The abrB gene of Bacillus subtilis is believed to encode a repressor that controls the expression of genes involved in starvation-induced processes such as sporulation and the production of antibiotics and degradative enzymes. Two such genes, spoVG, a sporulation gene of B. subtilis, and tycA, which encodes tyrocidine synthetase I of the tyrocidine biosynthetic pathway in Bacillus brevis, are negatively regulated by abrB in B. subtilis. To examine the role of abrB in the repression of gene transcription, the AbrB protein was purified and then tested for its ability to bind to spoVG and tycA promoter DNA. In a gel mobility shift experiment, AbrB was found to bind to a DNA fragment containing the sequence from -95 to +61 of spoVG. AbrB protein exhibited reduced affinity for DNA of two mutant forms of the spoVG promoter that had been shown to be insensitive to abrB-dependent repression in vivo. These studies showed that an upstream A + T-rich sequence from -37 to -95 was required for optimal AbrB binding. AbrB protein was also observed to bind to the tycA gene within a region between the transcription start site and the tycA coding sequence as well as to a region containing the putative tycA promoter. These findings reinforce the hypothesis that AbrB represses gene expression through its direct interaction with the transcription initiation regions of genes under its control. PMID- 2554318 TI - High-frequency T-DNA-mediated gene tagging in plants. AB - An insertion element [transferred DNA (T-DNA)], transferred by soil agrobacteria into the nuclear genome of plants, was used for induction of gene fusions in Arabidopsis thaliana, Nicotiana tabacum, and Nicotiana plumbaginifolia. A promoterless aph(3')II (aminoglycoside phosphotransferase II) reporter gene was linked to the right end of the T-DNA and transformed into plants along with a plasmid replicon and a selectable hygromycin-resistance gene. Transcriptional and translational reporter gene fusions were identified by screening for APH(3')II enzyme activity in diverse tissues of transgenic plants. The frequency of gene fusions, estimated by determination of the copy number of T-DNA insertions, showed that on average 30% of T-DNA inserts induced gene fusions in Arabidopsis and Nicotiana. Gene fusions were rescued from plants by transformation of the T DNA-linked plasmid and flanking plant DNA into Escherichia coli. By dissection of gene fusions and construction of chimeric genes, callus- and root-specific promoters were identified that showed an altered tissue specificity in the presence of a 3'-downstream-located 35S promoter. Transcript mapping of a gene fusion and expression of a non-frame transcriptional fusion of bacterial luciferase luxA and luxB genes demonstrated that dicistronic transcripts are translated in tobacco. PMID- 2554319 TI - Inhibition of Ca2+/calmodulin-dependent protein kinase II by arachidonic acid and its metabolites. AB - A variety of evidence indicates that activation of Ca2+/calmodulin-dependent protein kinase II (CaM-kinase II) in nerve terminals leads to enhanced neurotransmitter release. Arachidonic acid and its 12-lipoxygenase metabolite, 12 hydroperoxyeicosatetraenoic acid (12-HPETE), have been suggested to act as second messengers mediating presynaptic inhibition of neurotransmitter release. In the present study it was found that CaM-kinase II, purified from rat brain cortex, was inhibited both by arachidonic acid (IC50 = 24 microM) and by 12-HPETE (IC50 = 0.7 microM). Neither substance inhibited CaM-kinase I or III, protein kinase C, or the catalytic subunit of cAMP-dependent protein kinase. Specific inhibition of Ca2+/calmodulin-dependent protein phosphorylation by arachidonic acid was also demonstrated in intact synaptic terminals (synaptosomes) isolated from rat forebrain. These results suggest that arachidonate and its metabolites may modulate synaptic function through the inhibition of CaM-kinase II-dependent protein phosphorylation. PMID- 2554320 TI - Subunits of purified calcium channels: a 212-kDa form of alpha 1 and partial amino acid sequence of a phosphorylation site of an independent beta subunit. AB - Antibodies prepared against peptides CP2, CP4, and CP5, which occur within the first 1522 amino acid residues of the alpha 1 subunit of dihydropyridine sensitive skeletal muscle calcium channels, specifically recognized a 175-kDa form of the alpha 1 subunit in immunoblots and immunoprecipitation experiments. In contrast, antibodies prepared against peptide CP1, which represents the C terminal 18 amino acid residues predicted by cloning and sequence analysis of the alpha 1 subunit, recognized a minor, previously undescribed 212-kDa protein, which is the size predicted for the full length of the alpha 1 subunit from cDNA cloning [Tanabe, T., Takeshima, H., Mikami, A., Flockerzi, V., Takahashi, H., Kangawa, K., Kojima, M., Matsuo, H., Hirose, T. & Numa, S. (1987) Nature (London) 328, 313-318]. Both the 175-kDa and 212-kDa forms were phosphorylated by cAMP dependent protein kinase and both were present in isolated transverse tubule membranes. The 175-kDa form may arise from posttranslational proteolytic cleavage of the C terminus of the 212-kDa form of the alpha 1 subunit predicted by cDNA cloning and sequence analysis. Partial amino acid sequencing of the 54-kDa beta subunit of the calcium channel indicated this protein was not derived from the proteolytically cleaved C terminus of the alpha 1 subunit. This analysis identified a threonine residue in the sequence (Lys/Arg)-Arg-Pro-Thr-Pro of the beta subunit that was phosphorylated by cAMP-dependent protein kinase. Phosphorylation of this residue in the beta subunit may play a role in modulation of calcium channel function. Separate functional roles of the 175-kDa form of the alpha 1 subunit in excitation-contraction coupling and of the 212-kDa form in ion conductance are proposed. PMID- 2554321 TI - Specific recognition of the leader region of precursor proteins is required for the activation of translocation ATPase of Escherichia coli. AB - The ATP-hydrolytic activity of SecA protein is stimulated up to 100-fold by the translocation-competent precursor to outer membrane protein A (pro-OmpA) in conjunction with inner-membrane vesicles bearing active SecY [Lill, R., Cunningham, K., Brundage, L., Ito, K., Oliver, D. & Wickner, W. (1989) EMBO J. 8, 961-966]. This reaction is saturable, with Michaelis-Menten kinetics for an enzyme with two substrates, ATP and pro-OmpA, and is defined as translocation ATPase. Another precursor protein, pre-PhoE, is also a substrate for this translocation ATPase. Neither OmpA nor its synthetic leader peptide are effective substrates for translocation ATPase, suggesting that both domains of the complete precursor are necessary for the reaction. The leader peptide is a potent inhibitor and apparently competes with pro-OmpA for necessary binding sites on translocation ATPase. After a brief preincubation, the activity of translocation ATPase becomes resistant to inhibition by leader peptide, suggesting that the leader peptide is recognized at an early step in the protein translocation pathway. Our enzymological studies show that translocation ATPase recognizes and functionally binds the leader region of precursor proteins. PMID- 2554322 TI - Overreplication of the origin region in the dnaB37 mutant of Bacillus subtilis: postinitiation control of chromosomal replication. AB - When the Bacillus subtilis dnaB37 mutant, defective in initiation, is returned to permissive temperature after accumulation of initiation proteins at 45 degrees C, we have shown, by extensive DNA.DNA hybridization analysis, that the origin region is replicated in excess (approximately 2-fold). However, this replication is limited to a region of about 120-175 kilobases on either side of the origin. This has been confirmed by autoradiographic analysis of the overreplicated region. During the second round of synchronized replication at 30 degrees C, replication in fact appears to resume from the stalled forks on either side of the origin. We propose that in B. subtilis, in addition to a first level of control at the origin, a second level of control exists downstream of the origin in order to limit overreplication of the chromosome. These two controls might normally be tightly coupled. We suggest that the second level of control is exerted through the reversible inhibition of replisome movement at specific regions on either side of the origin. PMID- 2554323 TI - Molecular cloning and sequence of cDNA encoding polyoma medium tumor antigen associated 61-kDa protein. AB - Polyoma virus medium tumor antigen forms specific complexes with several cellular proteins; among these is a protein of approximately 61 kDa. With antibodies directed against medium tumor antigen, the 61-kDa protein was purified from human 293 cells that were infected with a hybrid adenovirus and overexpressed medium tumor antigen. The purified 61-kDa protein was partially digested with protease V8, and one of the protease V8 fragments was isolated and partially sequenced. The amino acid sequence information was used to design mixed oligonucleotide probes for screening a cDNA library from human placenta. A clone was isolated that hybridized with two separate probes; the clone contained an insert with an open reading frame for 589 amino acids. By in vitro translation of the transcript from this insert, a protein was generated that had the same size and yielded the same pattern of protease V8 fragments as the original 61-kDa protein. Its amino acid sequence reveals 15 repeats, the majority of which are 39 amino acids long. This protein bears no resemblance to proteins in the data bank that was searched. PMID- 2554324 TI - Detergent disruption of bacterial inner membranes and recovery of protein translocation activity. AB - Isolation of the integral membrane components of protein translocation requires methods for fractionation and functional reconstitution. We treated inner membrane vesicles of Escherichia coli with mixtures of octyl beta-D-glucoside, phospholipids, and an integral membrane carrier protein under conditions that extract most of the membrane proteins into micellar solution. Upon dialysis, proteoliposomes were reconstituted that supported translocation of radiochemically pure [35S]pro-OmpA (the precursor of outer membrane protein A). Translocation into these proteoliposomes required ATP hydrolysis and membrane proteins, indicating that the reaction is that of the inner membrane. The suspension of membranes in detergent was separated into supernatant and pellet fractions by ultracentrifugation. After reconstitution, translocation activity was observed in both fractions, but processing by leader peptidase of translocated pro-OmpA to OmpA was not detectable in the reconstituted pellet fraction. Processing activity was restored by addition of pure leader peptidase as long as this enzyme was added before detergent removal, indicating that the translocation activity is not associated with detergent-resistant membrane vesicles. These results show that protein translocation activity can be recovered from detergent-disrupted membrane vesicles, providing a first step towards the goal of isolating the solubilized components. PMID- 2554325 TI - A family of receptor-linked protein tyrosine phosphatases in humans and Drosophila. AB - To understand the regulation of cell proliferation by tyrosine phosphorylation, characterization of protein tyrosine phosphatases (PTPase; protein-tyrosine phosphate phosphohydrolase, EC 3.1.3.48) is essential. The human genes LCA (leukocyte common antigen) and LAR encode putative receptor-linked PTPases. By using consensus sequence probes, two additional receptor-linked PTPase genes, DLAR and DPTP, were isolated from Drosophila melanogaster. The extracellular segments of both DLAR and DPTP are composed of multiple immunoglobulin-like domains and fibronectin type III-like domains. The cytoplasmic region of DLAR and DPTP, as well as human LCA and LAR, are composed of two tandemly repeated PTPase domains. PTPase activities of immunoprecipitated LCA and LAR were demonstrated by measuring the release of phosphate from a 32P-labeled [Tyr(P)]peptide. Furthermore, the cytoplasmic domains of LCA, LAR, DLAR, and DPTP, expressed in Escherichia coli, have PTPase activity. Site-directed mutagenesis showed that a conserved cysteine residue is essential for PTPase activity. PMID- 2554326 TI - Thermal unwinding of simian virus 40 transcription complex DNA. AB - Two long-standing questions in the control of eukaryotic gene expression have been how the structure of transcribing chromatin compares with that of nontranscribing chromatin and how chromatin structure differs among various eukaryotic organisms. We have addressed aspects of these two questions by characterizing the rotational flexibility of the DNA of the simian virus 40 (SV40) transcription complex. When transcription complex samples are incubated with topoisomerase at 0 degrees C or 37 degrees C, the DNA of the 37 degrees C sample is unwound by 1.8 turns relative to that of the 0 degrees C sample. This amount of unwinding is similar to that observed for bulk, untranscribed SV40 minichromosome DNA, indicating that the chromatin structure of a transcribed gene resembles that of a nontranscribed gene in the degree of constraint that it imposes on its DNA. However, this amount of unwinding differs substantially from the value observed for yeast plasmid chromatin DNA, suggesting that yeast chromatin differs significantly from mammalian chromatin in this fundamental property. PMID- 2554327 TI - Receptor for acidic fibroblast growth factor is related to the tyrosine kinase encoded by the fms-like gene (FLG). AB - We have previously isolated a human gene from an endothelial cell cDNA library encoding a putative tyrosine kinase; we have designated this gene the fms-like gene (FLG). To analyze the gene product(s) of FLG, we have generated rabbit polyclonal antibodies directed against a synthetic peptide from FLG and used it to immunoprecipitate biosynthetically labeled FLG protein from a variety of human cell lines. These antibodies specifically recognized glycoprotein(s) of 100, 120, and 135 kDa with protein cores of 90 and 110 kDa. Acidic fibroblast growth factor (aFGF) stimulated tyrosine kinase activity of FLG in vitro and in living cells, suggesting that FLG encodes the membrane receptor for aFGF. Further supporting evidence came from cross-linking experiments on intact cells with the covalent cross-linking agent disuccinimidyl suberate and 125I-labeled aFGF as a specific probe. The cross-linked 125I-labeled aFGF-aFGF receptor complex was specifically immunoprecipitated with FLG antipeptide antibodies. It appears, therefore, that the receptor(s) for aFGF is related to the FLG gene product. PMID- 2554328 TI - Photolysis of a photolabile precursor of ATP (caged ATP) induces microsecond rotational motions of myosin heads bound to actin. AB - To test the proposal that ATPase activity is coupled to the rotation of muscle cross-bridges (myosin heads attached to actin), we have used saturation-transfer EPR to detect the rotational motion of spin-labeled myosin heads (subfragment 1; S1) bound to actin following the photolysis of caged ATP (a photoactivatable analog of ATP). In order to ensure that most of the heads were bound to actin in the presence of ATP, solutions contained high (200 microns) actin concentrations and were of low (36 mM) ionic strength. Sedimentation measurements indicated that 52 +/- 2% of the spin-labeled heads were attached in the steady state of ATP hydrolysis during EPR measurements. Five millimolar caged ATP was added to the actin-S1 solution in an EPR cell in the dark, with no effect on the intense saturation-transfer EPR signal, implying a rigid actin-S1 complex. A laser pulse produced 1 mM ATP, which decreased the signal rapidly to a brief steady-state level that indicated only slightly less rotational mobility than that of free heads. After correcting for the fraction of free heads, we conclude that the bound heads have an effective rotational correlation time of 1.0 +/- 0.3 microseconds, which is about 100 times shorter (faster) than that in the absence of ATP. To our knowledge, this is the first direct evidence that myosin heads undergo rotational motion when bound to actin during the ATPase cycle. It is likely that similar cross-bridge rotations occur during muscle contraction. PMID- 2554329 TI - MSI1, a negative regulator of the RAS-cAMP pathway in Saccharomyces cerevisiae. AB - We have previously demonstrated that the IRA1-encoded protein inhibits the function of the RAS protein in a fashion antagonistic to the function of the CDC25 protein in the RAS-cAMP pathway in Saccharomyces cerevisiae. In an attempt to identify genes involved in the regulation of this pathway, high-copy-number plasmid suppressors of the heat shock sensitivity of the ira1 mutation were isolated. One such suppressor, MSI1, was found to encode a putative protein of 422 amino acids that shows homology to the beta subunit of the mammalian guanine nucleotide-binding regulatory proteins. Overexpression of the MSI1 gene could suppress the heat shock sensitivity and the defect in sporulation caused by the ira1 and RAS2Val19 mutations but not those of the bcy1 mutation. Furthermore, the high level of intracellular cAMP in ira1 and RAS2Val19 cells was reduced by the MSI1 gene carried on a YEp-based plasmid. These results suggest that the MSI1 protein is a negative regulator of the RAS-mediated induction of cAMP in S. cerevisiae. PMID- 2554330 TI - Stimulation of the expression of osteogenic and chondrogenic phenotypes in vitro by osteogenin. AB - Osteogenin was recently purified and the amino acid sequences of tryptic peptides were determined. Osteogenin in conjunction with insoluble collagenous bone matrix induces cartilage and bone formation in vivo. To understand the mechanism of action of osteogenin, we examined its influence on periosteal cells, osteoblasts, fibroblasts, chondrocytes, and bone marrow stromal cells in vitro. Osteogenin stimulated alkaline phosphatase activity and collagen synthesis in periosteal cells. The cAMP response to parathyroid hormone in periosteal cells was increased by osteogenin. In primary cultures of calvarial osteoblasts, osteogenin stimulated alkaline phosphatase activity, the cAMP response to parathyroid hormone, and the synthesis of collagenous and noncollagenous proteins; however, cell proliferation was not affected. Osteogenin increased the production of sulfated proteoglycans in fetal rat chondroblasts and in rabbit articular chondrocytes. The present experiments demonstrate the significant influence of osteogenin in the stimulation of osteogenic and chondrogenic phenotypes in vitro. PMID- 2554331 TI - Spatial and temporal pattern of expression of the cellular retinoic acid-binding protein and the cellular retinol-binding protein during mouse embryogenesis. AB - Retinol (vitamin A) and retinoic acid are potent teratogens and also represent good candidates for normal morphogens during development. Their actions may be mediated by the cellular retinoic acid-binding protein (CRABP) and the cellular retinol-binding protein (CRBP). As a step towards understanding the possible function for CRABP and CRBP in morphogenesis, we have used in situ hybridization to analyze their expression during mouse development. Both CRABP and CRBP transcripts were detected at embryonic days 9.5-14.5. (i) In the nervous system, CRABP transcripts were found in the mantle layer of the dorsal spinal cord and hindbrain and in the marginal layer of the midbrain, whereas CRBP transcripts were found in the ependymal and mantle layer of the ventral spinal cord and of the forebrain as well as in the spinal nerves and the roof plate of the spinal cord. (ii) In the eye, CRABP is expressed in the retinal layer, and CRBP is expressed in both retinal and pigmented layers. (iii) In the craniofacial region, CRABP transcripts were found in the mesenchyme of the frontonasal mass and mandible, while CRBP transcripts were found in the mesenchyme of the nasolachrymal duct and surrounding the auditory vesicle. Two general conclusions can be made. First, all of the tissues that are known to be teratogenic targets of retinoic acid and retinol also express CRABP and CRBP transcripts. Second, the specific expression of CRABP and CRBP in numerous developing tissues indicates that these proteins may perform specific functions during morphogenesis of a broad variety of embryonic structures. PMID- 2554332 TI - Temporal expression and location of colony-stimulating factor 1 (CSF-1) and its receptor in the female reproductive tract are consistent with CSF-1-regulated placental development. AB - During pregnancy the mouse uterine epithelial synthesis of the mononuclear phagocyte growth factor designated colony-stimulating factor 1 (CSF-1) is regulated by female sex steroids. To study the role of CSF-1 in the pregnant female reproductive tract, the temporal expression and cellular sites of synthesis of CSF-1 and CSF-1 receptor (CSF-1R) mRNA were determined. CSF-1 mRNA, predominantly the 2.3-kilobase (kb) form, was first detected by in situ hybridization in uterine epithelium prior to implantation on day 3 and subsequently increased, reaching a peak at days 14-15. Its expression was restricted to the uterine epithelium at all stages of gestation and was not localized to areas of implantation. CSF-1R mRNA was first detected in maternal decidua at day 6. It was expressed in the decidua basalis during placentation, after which its expression declined. At day 7.5, trophectodermal cells also expressed CSF-1R mRNA; during placentation, it was found also in the diploid trophoblasts. The high level of CSF-1R mRNA expression by trophoblast giant cells was independent of their location around the conceptus. There was a differential distribution of CSF-1R mRNA expression in the mature placenta, with expression in the giant trophoblastic layer greater than spongiotrophoblastic layer greater than labyrinthine layer until term. Yolk sac cells also expressed low levels of CSF-1R mRNA. The coincidence of uterine CSF-1 mRNA expression and CSF-1 synthesis with both placental growth and CSF-1R mRNA expression in decidual cells and trophoblasts strongly implicates CSF-1 in the regulation of placental growth and differentiation. PMID- 2554333 TI - Amplification-control element ACE-3 is important but not essential for autosomal chorion gene amplification. AB - We have further characterized the cis-acting elements that control the amplification of the third chromosomal cluster of chorion genes in Drosophila melanogaster; these include the amplification-control element ACE-3 and four amplification-enhancing regions (AER-a to -d). We have used two types of deletions in the chorion cluster: the first was in vitro generated deletions of the ACE-3 region that were subsequently introduced into the germ line, and the second was deletions induced in vivo within a transposon at a preexisting chromosomal location, thus avoiding the complication of position effects. Some of the lines bearing deletions of either type showed amplification, albeit at drastically reduced levels. These unexpected results indicate that, despite its importance, ACE-3 is not essential for low-level amplification and that cis acting amplification elements are functionally redundant within the autosomal chorion replicon. PMID- 2554334 TI - The unexpected antitermination of gyrA-directed transcripts is enhanced by DNA relaxation. AB - We show that transcription originating at the gyrA promoter of Escherichia coli is less subject to termination at the lambda Toop terminator (22% read-through) than is transcription originating from either the galOP (1% read-through) or topA (3% read-through) control regions. Furthermore, termination of the gyrA transcripts decreases (60% read-through) upon relaxation of the template DNA. We believe that events related to stimulation of transcription at the gyrA promoter by DNA relaxation are responsible for the enhanced terminator read-through. PMID- 2554335 TI - A long interspersed repetitive element--the I factor of Drosophila teissieri--is able to transpose in different Drosophila species. AB - Long interspersed repetitive elements (LINEs) are transposable elements present in many species. In mammals they are difficult to study because most of them are defective and their transposition frequency is low. The I factor of Drosophila melanogaster is a LINE element that is particularly interesting because its transposition occurs at high frequency during I-R hybrid dysgenesis. This phenomenon occurs when males from the class of inducer strains are crossed with females from the class of reactive strains. Inducer strains contain several complete 5.4-kilobase I factors at various sites on the chromosomal arms. Reactive strains are devoid of complete I factors. Many results indicate that active I factors have invaded the D. melanogaster genome recently. To study the evolutionary history of I elements, we have cloned and sequenced a potentially active I factor from Drosophila teissieri. It is flanked by a target-site duplication and terminates at the 3' end by tandem repeats of the sequence TAA. When introduced into the germ line of a reactive strain of D. melanogaster by P element-mediated transformation, it is able to transpose and induces hybrid dysgenesis. This strengthens the hypothesis of a recent reinvasion of the D. melanogaster genome by active I factors giving rise to the inducer strains. They could have originated by horizontal transfer from another species. Such events also could occur for other LINE elements and might explain the spread of new variants in mammalian genomes. Moreover, the results give a further insight into I factor functional organization. PMID- 2554336 TI - DNA fingerprinting by sampled sequencing. AB - We describe a method for characterizing DNA segments that combines limited sequencing with size separation of restriction fragments. As part of a multistep procedure, 5' overhangs of unknown sequence are generated by cleavage with a class IIS restriction enzyme. After labeling of these ends by using dideoxynucleotides tagged with distinctive fluorescent dyes, the restriction fragments are analyzed by polyacrylamide gel electrophoresis and detection of fluorescent emissions using a commercially available DNA sequencer. The nucleotide-specific fluorescent signatures permit determination of the terminal sequence for each labeled end. The set of labeled fragments, characterized by both size and terminal sequence, constitutes a fingerprint that can be used to compare DNA segments for overlap or relatedness. The inclusion of terminal sequence data dramatically increases the information content of the fingerprint, making comparisons more reliable and efficient than those based upon size alone. PMID- 2554337 TI - Definition of a bacterial virulence factor: sialylation of the group B streptococcal capsule. AB - Sialylation of bacterial capsules has been proposed as an important virulence factor for several species of encapsulated pathogens, including group B Streptococcus. We have constructed a transposon mutant strain of type III group B Streptococcus that expresses a capsular polysaccharide differing from the wild type only in that the mutant strain's capsule lacks sialic acid. The mutant polysaccharide is antigenically identical to the capsular polysaccharide of type 14 Streptococcus pneumoniae, as predicted by the structures of the type III group B Streptococcus and S. pneumoniae polysaccharides. Loss of capsular sialic acid was associated with loss of virulence in the mutant strain in a neonatal rat model of lethal group B Streptococcus infection. These studies demonstrate directly that capsular sialic acid is a critical virulence determinant for type III group B Streptococcus and support the general hypothesis that surface sialylation aids pathogenic microorganisms in evading host defenses. PMID- 2554338 TI - Frequency-dependent release of peptide cotransmitters from identified cholinergic motor neurons in Aplysia. AB - We have investigated the release of two peptide cotransmitters from the terminals of a cholinergic motor neuron in Aplysia. Identified motor neuron B15 synthesizes the two small cardioactive peptides (SCP) A and B in addition to acetylcholine. A symmetrical pair of B15 neurons innervate symmetrical buccal muscles, termed I5, which are involved in generating biting movements. The amplitude of I5 contractions is enhanced by the SCPs. Intracellular stimulation of one B15 produces depletion of the SCPs from the stimulated muscle as compared to the unstimulated control muscle. Significant depletion requires either high-frequency stimulation or prolonged bursts at lower frequencies. A second cholinergic motor neuron, B16, also innervates I5 but does not synthesize the SCPs. Stimulation of B16 produced no depletion of the SCPs. Exogenous SCPs potently increase cAMP levels in the muscle. If depletion is a reflection of release, it should be possible to demonstrate an effect of B15 stimulation on muscle cAMP levels. Indeed, stimulation of B15 did elevate cAMP levels in I5. Stimulation of B16 had no effect on cAMP levels. Increases in cAMP were observed only when B15 was stimulated in a manner that would produce significantly facilitated acetylcholine release. This facilitation could be produced by increased stimulation frequency, longer burst durations, or shorter interburst intervals. However, B15 is capable of producing cholinergically mediated contractions with stimulation parameters that would not cause release of the SCPs. Thus, B15 appears to function as a purely cholinergic motor neuron when firing slowly, and as a cholinergic/peptidergic neuron when firing rapidly. PMID- 2554339 TI - Naloxone modulates body and organ growth of rats: dependency on the duration of opioid receptor blockade and stereospecificity. AB - Endogenous opioid systems (i.e., endogenous opioids and opioid receptors) act in the trophic regulation of biological development. Opioid antagonist paradigms have served to elucidate the nature of this relationship. If growth is mediated at the level of the opioid receptor, one would expect that this interaction would be stereospecific. This study shows that daily injections of 20-60 mg/kg (-) naloxone, given chronically throughout the preweaning period, depress body weight when monitored at Day 21. Opioid challenge experiments using nociceptive measures show that these dosages of (-) naloxone invoked an opioid receptor blockade for no more than 10-12 hr/day. A dosage of 100 mg/kg (-) naloxone, which blocked the opioid receptor for 12-16 hr/day, did not alter body weight in comparison to control levels. In subsequent experiments, 40 mg/kg (-) naloxone depressed body weight of 21-day-old rats, and the wet weights of the liver, spleen, thymus, heart, and triceps surae muscle from these animals were subnormal. A dosage of 40 mg/kg (+) naloxone did not alter growth. These results show that opioid action in regard to growth is stereospecific and dependent on the duration of opioid receptor blockade, providing additional evidence that endogenous opioid systems play an important role in developmental events. PMID- 2554340 TI - Voluntary ethanol intake of individually- or pair-housed rats: effect of ACTH or dexamethasone treatment. AB - The effect of ACTH or dexamethasone treatment on ingestion of 10% ethanol, 0.5 M NaCl and water was studied in individually- and pair-housed rats. Crowding or decreasing the amount of space per rat by increasing the number of rats per cage from 1 to 2, together with the associated increase in social interactions caused a large increase in ethanol intake. In pair-housed rats and in rats housed alone, ACTH treatment caused a large increase in Na intake but no change in ethanol intake. In pair-housed rats and in rats housed alone, dexamethasone treatment caused no change in either ethanol or Na intake. Thus, it would appear that the induction or maintenance of a high ethanol intake of rats during crowding, a presumed social stressor, can not be attributed entirely to either an increase in blood ACTH levels with the subsequent increase in glucocorticoid hormones or to a decrease in blood ACTH and natural glucocorticoid hormone levels. However, the possibility that ACTH and/or adrenocorticoid hormones, combined with other physiological or environmental factors, causes stressor-induced ethanol intake cannot be excluded. PMID- 2554341 TI - Effects of kappa-opioid receptor agonists and morphine on food intake and urinary output in food-deprived and nondeprived rats. AB - The effects of kappa-opioid receptor agonists, bremazocine, U-50, 488H and tifluadom and of a mu-opioid receptor agonist, morphine, on food intake and urinary output in food-deprived and nondeprived Sprague-Dawley rats was determined. In food-deprived animals, intraperitoneal administration of bremazocine at 0.1 mg/kg increased food intake but at 1.0 and 10.0 mg/kg doses decreased it. Tifluadom (0.1-10.0 mg/kg) had no effect on food intake. U-50,488H at 1.0 mg/kg increased food intake, whereas 10.0 mg/kg dose decreased the food consumption. In nondeprived rats, the kappa-opioid receptor agonists failed to produce any effect on food consumption. In food-deprived rats, all the three kappa-opioid receptor agonists increased the urinary output at the highest dose (10 mg/kg). In nondeprived rats similar effects as in food-deprived rats were observed except bremazocine increased urinary output at all the doses used. These results with kappa-opioid agonists may be related to either the existence of more than one population of kappa-opioid receptors or their differential actions at the opioid receptor types. PMID- 2554342 TI - Delta 9-tetrahydrocannabinol facilitates striatal dopaminergic transmission. AB - We examined the effects of delta 9-tetrahydrocannabinol (THC) on striatal dopaminergic neurons in rats. THC inhibited the uptake of 3H-dopamine (DA) into striatal synaptosomes. THC facilitated the release of endogenous DA but not dihydroxyphenylacetic acid (DOPAC) from striatal slices. The concentration of DA in the dorsolateral striatum was reduced by THC. We propose that THC may stimulate nigrostriatal dopaminergic neurotransmission mainly by inhibiting uptake of DA and by facilitating release of DA. PMID- 2554343 TI - Brain benzodiazepine receptors in fathead minnows and the behavioral response to alarm pheromone. AB - Fathead minnows (Pimephales promelas) exposed to 1 or 10 mg/l chlordiazepoxide showed normal alarm behavior during the presentation of alarm pheromone. Fish exposed to 20 mg/l drug, however, showed little or no behavioral alarm and did not appear sedated. A food extract stimulus presented after alarm pheromone led to a large foraging response by fish exposed to 20 mg/l chlordiazepoxide. Control fish or fish exposed to 1 to 10 mg/l drug showed less tendency to begin foraging. Exposure to 1, 10, or 20 mg/l chlordiazepoxide for 3 hr did not affect whole brain concentrations of tryptophan, 5-hydroxytryptamine, 5-hydroxyindoleacetic acid, tyrosine, or dopamine. The binding of [3H]diazepam to specific brain receptor sites (KD = 10 nM, estimated Bmax = 3.3 fmol/mg wet weight) could be displaced by chlordiazepoxide (IC50 = 1.6 microM). These results suggest that benzodiazepine receptors in the central nervous system of lower vertebrates may function in ways similar to those in mammals, i.e., in the modulation of behavior in anxiety-like states. PMID- 2554345 TI - DNA topoisomerases as potential targets of antiviral action. PMID- 2554344 TI - Breathhold duration and response to marijuana smoke. AB - Marijuana smokers are frequently observed to hold the smoke in their lungs for prolonged periods (10-15 sec) apparently in the belief that prolonged breathholding intensifies the effects of the drug. The actual influence of breathhold duration on response to marijuana smoke has not been studied. The present study examined the effects of systematic manipulation of breathhold duration on the physiological, cognitive and subjective response to marijuana smoke in a group of eight regular marijuana smokers. Subjects were exposed to each of three breathhold duration conditions (0, 10 and 20 sec) on three occasions, scheduled according to a randomized block design. A controlled smoking procedure was used in which the number of puffs, puff volume and postpuff inhalation volume were held constant. Expired air carbon monoxide levels were measured before and after smoking to monitor smoke intake. Typical marijuana effects (increased heart rate, increased ratings of "high" and impaired memory performance) were observed under each of the breathhold conditions, but there was little evidence that response to marijuana was a function of breathhold duration. PMID- 2554346 TI - Plasma corticosterone response in continuous versus discontinuous chronic heat exposure in rat. AB - Effects of chronic exposure (25 days) to continuous (CHE) or discontinuous (DHE) hot environment (34 degrees C) on growth rate, food intake and cortico-adrenal function were studied in adult male rats. Growth rate and food intake were 40 and 31% less respectively in CHE and 7 and 9% less respectively in DHE than in control (CE) animals. The adrenal response to heat (40 degrees C) was reduced by 54 and 82% in CHE and DHE rats respectively but the plasma corticosterone (B) response to exogenous ACTH and ether stress was not altered by chronic heat. The plasma B rhythm was altered only in CHE rats which exhibited a 40% decrease in amplitude and a delayed nocturnal recession of plasma B values. These results indicate that 1) CHE induced major alterations of behavior specially during the nocturnal period of the L-D cycle, 2) that the altered circadian B rhythm of CHE rats could be linked to changes in feeding behavior and to reduced metabolic activity, 3) and that CHE induced a poorer adaptation than DHE. PMID- 2554347 TI - The effects of adrenalectomy, corticotropin releasing factor and vasopressin on the sympathetic firing rate of nerves to interscapular brown adipose tissue in the Zucker rat. AB - The firing rate of the sympathetic efferent nerves to interscapular brown adipose tissue (IBAT) is lower in the obese rat compared with the lean rat. The present experiments show that adrenalectomy has no effect on nerve firing rate in the lean rat and a small but statistically nonsignificant effect in the obese rat. Injection of corticotropin releasing factor (CRF) into the IIIrd ventricle produced a dose dependent increase in the firing rate of the sympathetic nerves to interscapular brown adipose tissue (IBAT) in both lean and obese rats. The basal (unstimulated) level of firing was lower in the obese rat compared with the lean rat and remained significantly below lean values at each dose. The minimum dose of CRF to see an effect (125 ng) and the dose at which maximum effect on nerve firing rate was observed (500 ng) was similar in both genotypes. Injection of adrenocorticotropic hormone (ACTH) had no effect on nerve firing rate to IBAT. Central administration of vasopressin produced a significant increase in sympathetic firing rate to IBAT in both lean and obese rats. The temperature of IBAT was also significantly increased with vasopressin and the duration of the response was longer compared with CRF, but the minimum dose to see an effect was higher (2.5 micrograms). The response to vasopressin was greater in the obese rat compared with the lean rat but the maximum firing rate did not achieve levels observed in lean rats. Chronic infusion of CRF into the IIIrd ventricle of obese rats resulted in a reduction of food intake and body weight gain but IBAT mitochondrial GDP binding was unaltered by the treatment. These data are consistent with the hypothesis that the defect in the obese Zucker rat may be due to a glucocorticoid inhibition of CRF and/or vasopressin action in the CNS. PMID- 2554348 TI - The influence of maternal separation on plasma concentrations of ACTH, epinephrine, and norepinephrine in guinea pig pups. AB - Preweaning guinea pigs were placed into a test cage either alone or with their mother for 10, 30, or 90 min, or were placed into the cage alone for 24 hr. At 30 and 90 min, pups exhibited higher plasma levels of ACTH if alone than if with the mother. This effect was not apparent at 10 min, and ACTH levels of pups tested alone were no longer greater than baseline levels at 24 hr. These results provide further evidence that brief maternal separation can serve as a potent stimulus for activation of the hypothalamo-pituitary-adrenal system in this species. Plasma epinephrine and norepinephrine concentrations of pups tested alone were not significantly greater than those of pups tested with their mothers. But, pups tested alone for 10 and 30 min did show an elevation of epinephrine and norepinephrine over baseline levels. That is, the combined influence of separation and other aspects of the manipulation (e.g., exposure to novelty) evoked sympathetic responses. In addition, the number of vocalizations emitted by pups during the first 30 min of isolation in the test cage was positively correlated with concentrations of ACTH, epinephrine, and norepinephrine following 24 hr of this procedure, but not following 90 min or at baseline. Thus, the initial behavioral response appears to be predictive of the levels of both hypothalamo-pituitary-adrenal and sympathetic activity 24 hr after separation is initiated. PMID- 2554349 TI - Adrenal and behavioral responses of swine restricted to varying degrees of mobility. AB - Adrenal and behavioral responses in swine restricted to varying degrees of mobility were examined to determine what component may impose chronic stress. The components of restriction considered were the inability to turn around and/or move freely. For 5 to 6 weeks 32 pigs were restrained such that pigs A) could move freely; B) could not turn around or move freely; C) could not turn around but could move back and forth the same distance as pigs in A; or D) could turn around but not move freely. After 4 weeks no significant differences due to degree of restraint were found in the profile of serum cortisol concentrations, neutrophil:lymphocyte ratio and ambulatory abilities; however overall cortisol concentrations were higher in pigs that could turn around but not move freely. Increases in serum cortisol due to ACTH injection were higher in pigs that could turn around but not move freely. No alterations in the manner in which gilts stood up and layed down could be observed. Collectively these data suggest that swine adapt when mobility is restricted to not turning around and not moving freely but cannot when animals can turn around but not move freely. PMID- 2554350 TI - High affinity binding of the calcium channel blocker, (+)-[methyl-3H]PN200 110(3HPN) in rat brain. AB - PN200-110 is a recently introduced 1,4-dihydropyridine which has been demonstrated to be a potent calcium channel blocker. 3HPN has been shown to bind in a specific saturable manner to P2 fractions obtained from brain homogenates from male Sprague-Dawley rats. 3HPN binding was found to be temperature dependent. Specific 3HPN binding was maximal at 25 degrees C; binding decreased at 2 degrees C and 37 degrees C. The KD calculated from Scatchard analysis was 0.0943 +/- 0.0038 nM while the Bmax was found to be 109.1 +/- 2.3 fmol/mg protein. A concentration dependent inhibition of 3HPN binding by various cations was determined and found to be as follows: ZN2+ greater than La3+ greater than Rh3+, Al3+ greater than Co2+, Ni2+, Mn2+ greater than Ca2+, Mg2+ greater than Ba2+ greater than Sr2+. These results provide evidence for the existence of central high affinity dihydropyridine receptor sites in rat brain. PMID- 2554351 TI - Cyclic-AMP in human lung preparations. AB - Cyclic-AMP levels in unaffected and affected cancerous portions of human lung preparations were determined by measuring displaced 3H-cyclic-AMP from the specific binding protein by unlabeled ligand. The levels of cyclic-AMP in unaffected portions of human lungs ranged between 52-116 pmoles/g protein. Significantly higher levels of cyclic-AMP were found in lung samples affected with squamous cell carcinoma, adenocarcinoma and malignant melanoma with the mean values being 251, 290, and 509 pmoles/g protein, respectively. On the other hand, a decrease in the level of cyclic-AMP to 39 pmoles/g protein was observed in portions of lungs affected with granuloma. These results suggest that the level of cyclic-AMP in lung tissue may reflect the malignant or benign nature of the pulmonary disease. PMID- 2554352 TI - Mechanisms of general anesthesia: brain regional responses to baclofen. AB - The GABAB agonist baclofen is reported to produce general anesthesia when administered either centrally into the lateral ventricles of rats or peripherally to mice. Previously we demonstrated that beta-endorphin given intracerebrally produces anesthesia in rats, a response localized to sites in or adjacent to the inferior third and fourth ventricles. In order to compare the anatomical localization of these two anesthetic responses, we administered baclofen into the inferior or superior lateral or third ventricles, the aqueduct, or fourth ventricle in rats. Although 10 micrograms baclofen infusions into several regions caused loss of the righting reflex, in no case did animals exhibit an unconscious state which satisfied strict criteria of anesthesia. Infusions of 20 micrograms into the inferior third and fourth ventricles elicited seizures followed by a postictal depression. Although unresponsive to some stimuli, these animals showed no impairment in the corneal reflex. Since this dose was often lethal, higher doses not tested. Baclofen, given to mice intraperitoneally at doses of 25, 50, or 75 mg/kg, failed to elicit strictly defined anesthesia, although, to varying degrees, animals exhibited analgesia, loss of the righting reflex, and loss of behavioral responses to loud sounds. Animals continued to show motor responses when handled and retained corneal reflexes. Baclofen does not evoke an unconscious anesthetic state when administered centrally or systemically, emphasizing the need for strict criteria to define general anesthesia and to categorize drugs that promote this state. PMID- 2554353 TI - Interactive effects of chronic phencyclidine and delta 9-tetrahydrocannabinol on spermatogenesis in mice. AB - In this study, the combined effects of chronic phencyclidine (PCP) and delta 9 tetrahydrocannabinol on spermatogenesis in mice were examined. Mice were treated with THC (50 mg/kg, PO) and PCP (15 mg/kg, IP) alone or in combination for 16 days and with PCP alone for 35 days. THC had a significant effect on spermatogenesis and decreased the number of all germinal cells. PCP, on the other hand, affected all germinal cells except spermatids after 35 days of treatment. Combination of THC and PCP treatment caused a significant decrease in resting and pachytene spermatocytes. Similarly, combination of these two drugs caused a significant increase in cauda epididymal abnormal sperms. These results suggest that THC and PCP may cause greater disruption in spermatogenesis when they are abused together. PMID- 2554354 TI - Effects of delta-9-tetrahydrocannabinol on in vitro energy substrate metabolism in mouse and rat testis. AB - In this in vitro radiorespirometric study, the effects of delta-9 tetrahydrocannabinol (THC) on glucose and fructose metabolism in the rat and mouse testes were compared. In the rat testis, THC caused a dose-dependent decrease of 15, 18, 20 and 25% in glucose metabolism with 0.1, 0.2 and 0.3 mM concentrations. Similarly, a 39% inhibition in fructose metabolism was observed when rat testes were exposed to 0.3 mM THC. The comparative studies with mouse testis indicated that 0.1, 0.2 and 0.3 mM THC produced a 4, 12 and 30% decrease in glucose utilization where as a much greater decrease of 13, 30 and 41% in fructose utilization was observed with the same doses of THC. On the basis of these observations in these two animal models, it is concluded that THC has the ability to inhibit energy substrates utilization in the rat and mouse testis. This inhibition of cellular energetics may, in part, be responsible for most of the gonadal effects of THC. PMID- 2554355 TI - Maternally-mediated neonatal lithium-cesium interaction in the mouse. AB - The effect of maternal exposure to LiCl, CsCl or both salts in the weaning and developing offspring mice was studied on selected organ weights, hepatic and cardiac dehydrogenase enzymes. The concentration of alkali metal used in maternal drinking fluid during pregnancy and breast-feeding did not produce taste aversion and therefore approximate equal consumption was assured. Maternal exposure to either alkali metal reduced brain and testis weights of the developing offspring mice compared to controls. This suggests a delayed toxic effect on the CNS and endocrine organs. Coadministration of both salts negated this effect. The maternal neonatal Li-mediated increases of weanling spleen weight and the reduction of testis weight of developing offspring mice by Li or Cs were not evident when both alkali metals were given in combination. The combined maternal exposure to both Li and Cs salts also negated the induction of offspring mouse liver alcohol dehydrogenase produced by either alkali metal alone. Likewise, the induction of developing mouse heart lactate dehydrogenase isoenzyme (LDH5) by maternal exposure to LiCl was no more apparent by the combined Li and Cs treatment. These data suggest a Li+-Cs+ interaction in the offspring mouse due to maternal exposure to these alkali metals during pregnancy and breast-feeding periods. The results also suggest that both alkali metals most probably have been delivered to the suckling pups and some of their toxic effect was retarded. PMID- 2554356 TI - Reduced natural killer cell activity in major depression: neuroendocrine implications. AB - Natural killer cell activity (NKCA) was significantly reduced in a group of depressed patients, melancholic subtype, compared to sex- and age-matched controls. Corticotropin and cortisol values were significantly higher in the depressed subjects than in the controls, but no correlation between high hormone levels and low immunological activity was found in the patients. PMID- 2554357 TI - Reduction of immunoreactive ACTH in plasma following intravenous injection of delta sleep-inducing peptide in man. AB - Eleven healthy male volunteers, ages 25-39 years, received a single dose of synthetic delta sleep-inducing peptide (DSIP) (25 nmol/kg BW) or saline intravenously in a randomized cross-over, double-blind study. The concentrations of neuropeptides related to the hypothalamic pituitary-adrenal (HPA) axis and cortisol were examined in serial plasma samples. In addition, cortisol and monoamine metabolites were determined in urine. A significant reduction of ACTH like immunoreactivity (ACTH-LI) in plasma was detected for at least 3 hr after the DSIP injection, compared to the control subjects, in whom a slightly elevated concentration of ACTH-LI occurred. Plasma cortisol levels were unaffected and followed the normal diurnal decline. No differences in urinary cortisol or monoamine metabolite concentrations occurred between the two groups. The results indicate an inhibitory action of DSIP on ACTH secretion in man, as previously suggested by animal experiments. PMID- 2554358 TI - Neuroendocrine evidence of deranged noradrenergic activity in chronic migraine. AB - Migraine is a psychobiological disorder in which a recurrent failure of opioid and adrenergic systems might occur, as plasma and CSF studies suggest. In order to elucidate the relationship between noradrenergic and opioidergic functions, the plasma beta-endorphin (beta-EP) response to clonidine and the cortisol response to dexamethasone were evaluated together in 25 patients suffering from migraine without aura, and with chronic tension headache (MTH). Baseline beta-EP plasma levels and beta-EP response to clonidine were significantly lower in MTH subjects than in controls, suggesting a postsynaptic hypothalamo-pituitary impairment. Forty-four percent of the MTH subjects showed either a lack of suppression of plasma cortisol following dexamethasone administration, or basal cortisol concentrations higher than controls and suppressors, suggesting a disinhibition of the hypothalamopituitary-adrenal (HPA) axis. An inverse correlation was found between pain severity and beta-EP secretion induced by clonidine (delta max), and no relationship was found between beta-EP and mood. These data suggest a failure of central noradrenergic activity, or perhaps an impaired secretion of beta-EP not related to HPA axis hyperactivity or to affective state. PMID- 2554359 TI - Psychoneuroendocrine effects of methadone maintenance. AB - A variety of neuroendocrine and psychiatric dysfunctions have been demonstrated in humans maintained on opiates, but both have not previously been examined in the same population. We performed a series of neuroendocrine challenge tests in men participating in a methadone maintenance clinic and in normal controls. Psychiatric diagnoses were made with DSM-III Criteria, using the Diagnostic Interview Schedule, and subjects also completed the Symptom Checklist. Our results in the methadone group suggest (a) near-maximal stimulation of prolactin secretion, with a blunted prolactin response to insulin hypoglycemia, (b) mild suppression of cortisol levels, but an exaggerated cortisol response to stimulation, (c) a delayed and inhibited insulin response to food ingestion with resulting mild hyperglycemia, (d) low body weight, but elevated calorie ingestion, and (e) inability to concentrate urine when dehydrated, which was partially corrected by administration of arginine vasopressin. Phobic disorder was associated with a lower prolactin response to both inhibitory and stimulatory challenges. Depression did not appear to be related to the increased cortisol response to stimulation. These results suggest several potentially fruitful areas for future investigation, including the prolactin system and anxiety disorders, nutrient ingestion and metabolism, and posterior pituitary function. PMID- 2554360 TI - Pachygyriclike changes: topographic appearance at MR imaging and CT and correlation with neurologic status. AB - Studies of 23 pediatric patients with pachygyriclike changes (PLCs) examined with computed tomography (CT) and magnetic resonance (MR) imaging were reviewed to determine topographic patterns and correlate them with various clinical syndromes and degrees of neurologic impairment. Three types of topographic distributions were identified: unilateral, diffuse, and bilateral nondiffuse (of which eight of 10 showed frontotemporal predominance). PLCs were an isolated finding in seven patients, were associated with various congenital syndromes in nine patients, and were associated with congenital infection in seven patients, six of whom showed marked white matter abnormalities. Although most patients had severe developmental delay, three with nondiffuse PLCs had less severe impairment, permitting less required care. PMID- 2554361 TI - Differentiation of radial scar from scirrhous carcinoma of the breast: mammographic-pathologic correlation. AB - Radial scar, a sclerosing ductal breast lesion characterized by an irregular stellate pattern of epithelial proliferation around a central fibroelastic core, may be confused histologically with scirrhous carcinoma of the breast. Mammographic features used to distinguish these two entities were found unreliable in a retrospective review of 255 consecutive stellate lesions. Of 73 nonpalpable carcinomas, fourteen (19%) had radiographic features of radial scar. Only the presence of microcalcifications in 11 of those patients helped the authors distinguish carcinoma from radial scars. Four of nine biopsy-proved radial scars had a dense central region, simulating the appearance of scirrhous carcinoma. Stellate lesions with radiolucent centers should be considered suggestive of carcinoma, particularly if associated with microcalcifications. PMID- 2554362 TI - Dacryocystography: comparison of water-soluble and oil-based contrast agents. AB - Dacryocystography has been widely used in the assessment of the nasolacrimal duct system, particularly in patients with epiphora. Our study was undertaken to evaluate image quality and level of patient discomfort during examinations with water-soluble contrast agents (iohexol [Omnipaque 240], iopamidol [Isovue 200 and 300], and 52.7% diatrizoate meglumine and 26.9% iodipamide meglumine [Sinografin]) compared with the iodized oil-based contrast agent Lipiodol. Fifty five dacryocystograms were obtained from 41 consecutive patients. The procedure was performed first with a water-soluble contrast agent, then repeated with Lipiodol. A distention technique was used with conventional radiography. Patients were asked to evaluate their level of discomfort (none, mild, moderate, severe). The images were evaluated separately by two radiologists, blinded to which water soluble agent was employed, and the images were graded on a five-point scale. Images obtained with Lipiodol were significantly better than those with other agents (P less than .02), and image quality deteriorated as iodine concentration decreased. Use of Isovue 300 and Sinografin produced significantly more patient discomfort (P less than .03) than the use of other agents. The authors conclude that, in most instances, Lipiodol is the contrast agent of choice with regard to both highest level of patient comfort and greatest conventional radiographic image quality among the agents compared. PMID- 2554363 TI - Effects of repeated trazodone administrations on serotonergic neurotransmission: biochemical studies. AB - 1. Repeated administrations of trazodone as well as imipramine or mianserin (10 mg/kg i.p. twice daily for 3 weeks) attenuated the norepinephrine (NE) stimulation of adenylate cyclase studied in brain minces. Therefore trazodone shares with "tricyclic" (imipramine) and "atypic" (mianserin) antidepressants the capability to modulate the beta-adrenergic function. 2. Daily treatments with imipramine or trazodone enhanced the Vmax of neural uptake of serotonin (5HT) in minces prepared from rat frontal cortex; in contrast mianserin failed to modify the [3H]-5HT uptake. 3. Repeated administrations of imipramine but not of trazodone or mianserin reduced the maximum number of [3H]-imipramine recognition sites which are located on serotonergic axon terminals. 4. Differently, only repeated administration of trazodone decreased Bmax values of [3H]-mianserin binding sites which are located on membranes innervated by serotonergic neurons. Moreover trazodone did not change the number or affinity of 5HT2 receptors either after single or repeated administrations; in contrast even a single administration with mianserin or repeated administrations with imipramine down regulated [3H]-ketanserin specific binding in membranes prepared from the frontal cortex. 5. Our observations therefore suggest that trazodone, imipramine or mianserin exerts similar effects on the adenylate cyclase system, by acting on a interneuronal loop which links serotonergic and noradrenergic transmission function. However, its exact mechanism of action, in part resembling both tricyclic and atypic depressants, requires further examination. PMID- 2554364 TI - Effect of captopril or enalapril on renal prostaglandin E2. AB - Since one of the hypotensive mechanisms of angiotensin-converting enzyme inhibitor (ACEI) has been suggested to be mediated through the renal kinin prostaglandin (PG) axis, the present study was designed to investigate the effect of captopril (C) or enalapril (E) on renal PGE2 excretion or synthesis. Wistar male rats (BW 200-250 g) were given orally captopril at 30 mg/kg/day or enalapril at 10 or 30 mg/kg for one week. Before and after ACEI, blood pressure (tail cuff method) as well as PRA and urinary PGE2 excretion was determined. Renopapillary slices were obtained from some of the rats including controls and incubated to determine PGE2 synthesis. C or E administration resulted in a blood pressure decrease of 21 to 36 mm Hg with an increase in PRA. Urine volume and sodium excretion increased after daily treatment with C or E at 30 mg/kg. Urinary PGE2 excretion increased 1.4-fold in response to C, but not to E. Papillary PGE2 synthesis demonstrated a marked decrease 2 h after in vivo administration of either ACEI compared to controls. However, when C or enalaprilat was added in vitro to renal slices obtained from controls, only C at 10(-5) M showed a significant 2-fold increase in renal PGE2 synthesis. These results suggest that (1) renal PGE2 synthesis may be dependent on circulating angiotensin II. (2) C, but not enalaprilat, has a direct stimulatory effect on renal PGE2 synthesis and (3) renal PGE2 may not be involved very much in the hypotensive effect of ACEI. PMID- 2554365 TI - Peritoneal fluid eicosanoids in chronic pelvic pain. AB - Products of the cyclooxygenase and lipoxygenase pathways of arachidonic acid metabolism were estimated in the cul-de-sac fluid from patients with endometriosis, pelvic adhesions and normal laparoscopic examinations, with and without chronic pelvic pain. No correlation between the symptoms, underlying diagnoses, and the concentrations of eicosanoids were observed. PMID- 2554366 TI - 1-[4-[3-[4-[bis(4-fluorophenyl)hydroxymethyl]-1- piperidinyl]propoxy]-3 methoxyphenyl]ethanone(AHR-5333): a selective human blood neutrophil 5 lipoxygenase inhibitor. AB - In this study we report the in vitro inhibition of leukotriene synthesis in calcium ionophore (A23187)-stimulated, intact human blood neutrophils by AHR 5333. The results showed that AHR-5333 inhibits 5-HETE, LTB4 and LTC4 synthesis with IC50 values of 13.9, 13.7 and 6.9 microM, respectively. Further examination of the effect of AHR-5333 on individual reactions of the 5-lipoxygenase pathway (i.e. conversion of LTA4 to LTB4, LTA4 to LTC4, and arachidonic acid to 5-HETE) showed that this agent was not inhibitory to LTA4 epoxyhydrolase and glutathione S-transferase activity in neutrophil homogenates. However, conversion of arachidonic acid (30 microM) to 5-HETE was half maximally inhibited by 20 microM AHR-5333 in the cell-free system. The inhibition of LTB4 and LTC4 formation in intact neutrophils by AHR-5333 appears to be entirely due to a selective inhibition of 5-lipoxygenase activity and an impaired formation of LTA4, which serves as substrate for LTA4 epoxyhydrolase and glutathione-S-transferase. AHR 5333 did not affect the transformation of exogenous arachidonic acid to thromboxane B2, HHT and 12-HETE in preparations of washed human platelets, indicating that this agent has no effect on platelet prostaglandin H synthase, thromboxane synthase and 12-lipoxygenase activity. The lack of inhibitory activity of AHR-5333 on prostaglandin H synthase activity was confirmed with microsomal preparations of sheep vesicular glands. PMID- 2554367 TI - [In vitro investigations of Candida strains for itraconazole]. AB - Results of studies in vitro of the susceptibility of 102 strains of Candida type (88--Candida albicans, 7--Candida intermedia, 3--Candida parapsilosis, 1--Candida stellatoidea, 3--Torulopsis candida), to itraconazole have been reported. The strains were isolated from lesions of the skin and mucous membranes. The strains tested showed no resistance to this drug. Itraconazole concentrations inhibiting completely the growth of the yeast-like fungi ranged from 0.02 to 35.0 micrograms/ml of a culture medium. Strains of Candida albicans have shown the least susceptibility, most Candida intermedia. Changes in sensitivity (MIC) of some strains of Candida type for itraconazole after 48 and 72-hrs periods of incubation were very little. PMID- 2554368 TI - [Opportunistic viral infections in AIDS]. AB - Authors pointed on etiologic connections between particular clinical syndroms in AIDS and microbial, parasitic and viral agents belonging to different taxonomic groups. Participation of viruses in etiologic, immunology, pathology and importance for chemotherapy pointing on proved and probable relations, were analysed. Valuable observations related with etiologic and opportunistic viral mechanisms of coinfections were mentioned, as well as main diagnostic aspects. References limited up to the most recent papers, according to authors opinion, scientifically and practically most important. PMID- 2554369 TI - [Respiratory viral infections in young children 1985-1988]. AB - In the period from May 1985 to June 1988 the authors (using the immunofluorescence method) examined 848 children aged 0-2 years hospitalized due to infections of respiratory tracts in the II Clinic of Pediatrics of the Pediatrics Department of the II Medical Faculty in the Medical Academy in Warsaw. The most frequent causes of infections in respiratory tracts were virus RS (21.1% of patients) and virus of parainfluenza type 3 (6.5% of patients). An increase in viral respiratory infections took place every year between early autumn and late spring. Virus RS was permanently present in the population examined, though significant increases in the number of children infected by that virus appeared from March to May and from October to January in every year of the examination. An increased incidence of type 3 parainfluenza virus infections usually appeared in September. Among the children examined, the authors also found 25 cases of simultaneous infections by two different viruses. The most frequent combination of infecting viruses were virus RS and virus of parainfluenza type 3. PMID- 2554370 TI - Laxative abuse in eating disorders. PMID- 2554371 TI - Factors influencing outcome of definitive radiotherapy for localized carcinoma of the prostate. AB - Definitive radiation therapy was administered to 577 patients with histologically confirmed carcinoma of the prostate localized to the pelvis. Results of therapy and prognostic factors are evaluated and compared with other reports. All patients were followed for a minimum of 3 years, the median period of observation is 6.5 years. The disease-free survival in stages A2 and B was 78% at 5 years and 60% at 10 years; in stage C, 60% at 5 years and 36% at 10 years. The overall actuarial survival in stage B patients was 76% at 5 years and 58% at 10 years, which is similar to the life expectancy of a comparable cohort of normal males. In stage C, the actuarial survival was 65% at 5 years and 38% at 10 years, approximately 15% below the normal life expectancy for a similar cohort. Histological differentiation of the tumor had a significant impact on survival, poorly differentiated tumors showing 20% lower survival rates, most likely related to a higher incidence of distant metastases. Age was not found to be a significant prognostic factor in patients with stage B tumors; however, in stage C, the 5-year disease-free survival was 43% in patients younger than 60 and 53% in those older than 60 years (p = 0.04). In stage B, patients who had control of the pelvic tumor exhibited a 80% actuarial 5-year survival and 60% at 10 years. This compares with an actuarial survival of 30% at 5 and 10 years when there was evidence of pelvic recurrence alone or combined with distant metastases. In stage C patients with pelvic tumor control, actuarial survival was 81% at 5 years and 50% at 10 years, in comparison with 20% 5-year and 10% 10-year survival when pelvic recurrence or distant metastases or a combination of both occurred. The method of diagnosis [transurethral resection of the prostate (TURP) versus needle biopsy] did not appear to be a major prognostic factor of survival or patterns of failure, except in patients with stage C moderately differentiated tumors. The interpretation of the lymphangiogram (normal, suspicious or abnormal) did not correlate with survival or tumor control. The pathologic status of the lymph nodes established at lymphadenectomy did not appear to relate to survival for patients with stage B disease.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2554372 TI - Autoradiographic localization of vasoactive intestinal polypeptide receptors in the rat mesenteric vascular tree. AB - By the use of combined in vitro radioreceptor binding and autoradiographic techniques, we analyzed the pharmacological properties and the anatomical localization of the vasoactive intestinal polypeptide (VIP) receptor in rat superior mesenteric artery and in medium and small mesenteric artery branches. 125I-VIP was bound by sections of rat superior mesenteric artery in a manner consistent with the labeling of specific VIP receptors, with Kd and Bmax values of 0.23 nM and 0.71 pmol/mg protein respectively. Inhibition of 125I-VIP binding with VIP and related peptides gives the following rank order of potency: VIP greater than peptide histidine methionine greater than secretin. Light microscope autoradiography reveals specific VIP binding sites within the medial layer of superior mesenteric artery and its branches. Medium and small sized vessels are richer in 125I-VIP binding sites than the larger ones. PMID- 2554373 TI - [Mother and child stress parameters during cesarean section with general and peridural anesthesia]. AB - This study compared maternal and fetal stress responses during cesarean section in either general anesthesia (GA) or epidural anesthesia (EA). Ten patients received GA with thiopental induction, intubation, and controlled ventilation with nitrous oxide and oxygen. After delivery, anesthesia was supplemented with fentanyl 0.2-0.3 mg. Ten patients received EA via catheter, using bupivacaine 0.5%, and prilocaine or lidocaine 1%. Maternal mean arterial pressure (MAP), HR, and plasma concentrations of epinephrine and norepinephrine (by HPLC/ECD), ADH, ACTH and cortisol (by RIA) were determined before and after induction, after delivery, at the end of the operation and 30 minutes postoperatively. Fetal catecholamine levels in umbilical artery blood were measured immediately after delivery. In addition, fetal blood gas analyses and Apgar scores were compared. Fetal epinephrine was slightly increased in the EA group (EA 132 pg/ml, GA 52 pg/ml). Norepinephrine was similar in both groups (EA 1.218 pg/ml, GA 1.124 pg/ml). Blood gas analyses and Apgar scores were also comparable. A negative correlation was found between norepinephrine and pH values in fetal umbilical artery blood (P = 0.01). Maternal epinephrine levels were lower under EA and below the normal range (EA 23 pg/ml, GA 77 pg/ml, P = 0.002); levels increased during GA and decreased during EA (P = 0.01). No statistical differences were seen in maternal norepinephrine (EA 206 pg/ml, GA 354 pg/ml). MAP was lower during EA (group levels EA 81 mmHg, GA 95 mmHg, P = 0.0002) and HR was higher during GA (group levels EA 89/min, GA 104/min, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554374 TI - [CT and MRT of vertebral hemangiomas]. AB - A retrospective comparative study of CT and MRI was carried out involving 38 vertebral haemangiomas; this revealed a typical signal pattern on MRI from benign lesions. It consists of a hyper-intense signal from the bone marrow affecting the T1/T2 sequences; this may be focal or involve the entire vertebral body. These characteristic signals were compared with CT images of the spine. The areas of bone that produce the high intensity signals on MRI appear on CT as spongy patterns with hypertrophic trabeculae surrounding mostly areas with negative absorption values. An analysis of the changes in the spongiosa has revealed three clearly defined types. The signals derived from haemangiomas extending beyond the bone have an intensity of normal spongiosa; this corresponds with an absence of fat, as demonstrated by CT. Extra-osseous components have low intensity T1 signals that increase in T2 sequences. PMID- 2554375 TI - [A computed tomographic analysis of vertebral rotation before and after surgical correction of idiopathic scoliosis]. AB - Preoperative and postoperative computer tomographs in 24 patients suffering from idiopathic scolioses who were treated by CD (Cotrel-Dubousset) surgery, showed a mean correction of the rotatory angle in the apex vertebra amounting to 28.5% in relation to the anterior midline of the body, and 23.8% in relation to the sagittal plane. Best results were seen in lumbar curves and in the lumbar curves of double major curves. In thoracic curves only minor corrections were observed. CD instrumentation is the first dorsal procedure that allows a three-dimensional correction of the scoliotic spine. PMID- 2554376 TI - [Ultrasonic anatomy and study technic of the normal carpal tunnel and the distal median nerve]. AB - The wrists of 16 normal volunteers were examined via high-resolution sonography with special reference to the carpal tunnel. In all cases the surfaces of the carpal bones, the flexor retinaculum, the tendons, the vessels, (Aa. radialis and ulnaris) and the nerves (Nn. medianus and ulnaris) were observed. The sonographic characteristics of the median nerve are discussed and the possibilities of identification are demonstrated. It appears that the echogenicity of the nerve depends strongly on the angle of the ultrasound beam. PMID- 2554377 TI - [The diagnosis of salivary gland lipoma]. AB - 8 patients with 9 lipomas of the salivary gland were reviewed retrospectively. The clinical diagnosis, the sonomorphology and the appearance in CT were analysed. By means of palpation none of the lipomas was specifically diagnosed. Usually sonography is the first imaging method in superficial salivary gland tumors. Therefore the knowledge of the sonomorphology is important for indicating CT. Sonographically a lipoma of the salivary gland should be suspected, if an oval shaped, slightly hypoechoic tumor with a typical feathered appearance and good compressibility is visualized. All lipomas of the salivary glands could be delineated sonographically. Due to their negative homogeneous attenuation values lipomas can be diagnosed specifically in CT. This enables the head and neck surgeon to avoid an operation in patients with no or minor complaints. PMID- 2554378 TI - [Determination of the value of conventional and computer-assisted tomography in examining the thoracic organs]. AB - We compared the results of CT and conventional tomography examinations of the thorax of 137 patients with each other and with the final results. In the diagnosis of bronchial carcinomas we found no significant differences between the two methods. In all other investigated items CT proved to be superior to conventional tomography, especially of the evaluation of mediastinum (90% vs. 35%). We conclude that CT is the method of choice in the staging of intrathoracic tumours and especially in the evaluation of hilar and mediastinal masses and there is no need for conventional tomography in these cases. PMID- 2554379 TI - [The value of MRT of the thorax in congenital tracheal stenoses]. AB - MRI of the thorax was performed in 24 children aged between six weeks and five years, in whom a tracheal stenosis had been demonstrated by bronchoscopy. Since bronchoscopy can only demonstrate the interior of the trachea, various imaging methods, such as CT and angiography, were used to demonstrate the topography and cause of the tracheal stenosis. MRI has shown that the most common cause of a stenosis in the central section of the trachea is focal compression by an aberrant brachiocephalic trunk (10 cases). Stenosis of the distal trachea could be due to anomalies of the aortic arch (5 cases), a dilated pulmonary artery (4 cases) or a soft tissue mass (3 cases). In all these patients, MRI was greatly superior to the conventional methods. By using a special technique, MRI made it possible to clarify the cause and localisation of a tracheal stenosis by a non invasive examination. PMID- 2554380 TI - [Intraoperative sonography. Experiences and indications in upper abdominal laparotomy]. AB - Intraoperative ultrasonography should be a mandatory diagnostic procedure in "occult insulinoma" of the pancreas, in partial liver resection for primary or secondary malignomas, and in different findings by preoperative diagnostic and intra-operative situs. PMID- 2554381 TI - [Quantitative studies on the influence of connective tissue and fatty structures on the ultrasonic image of the liver]. AB - Using a computerized ultrasound system, 63 autopsied livers were examined. A quantitative description of the ultrasound image was made by using statistical parameters from pattern recognition algorithms. As a reference a chemical/morphometrical classification into normal, diffuse and regional fatty infiltration and fibrosis/cirrhosis was performed. Diagnostic accuracy varied from 77% to 92%. Ultrasound tissue characterisation showed an influence of fatty and connective tissue structures on image parameters. This supplies evidence for the fact that computerised ultrasound examination yields more information from the ultrasound image than the normal observer. This may be the cause of an increased diagnostic accuracy when using computerized ultrasound systems, especially in the case of fibrosis/cirrhosis. PMID- 2554382 TI - [Computer-assisted ultrasonic B-scan texture analysis of experimental liver tumors]. AB - Primary liver cancer has been induced experimentally in female Wistar rats by diaminoazobenzene intoxication or subcapsular liver implantation of Novikoff cell suspensions. The ultrasonic B-scan image of these malignant tumours are described via statistical texture parameters in respect of image brightness, microtexture and macrotexture. A discrimination of the tumour texture profiles is possible with an accuracy of about 80%. There are correlations between ultrasonic image parameters and histomorphological tissue components but there is no specific link between the histological tumour-type and these B-scan data. PMID- 2554383 TI - [Relaxation time measurements in the differential magnetic resonance tomographic diagnosis of liver tumors]. AB - T1-weighted IR sequences and T2-weighted SE sequences and combined SE/IR sequences were carried out in 62 patients with malignant and 50 patients with benign liver tumours and the T1 and T2 relaxation times were measured. IR sequences proved the most sensitive method for imaging focal liver lesions and providing good contrast between liver tissue and tumour. For lesions above 1.5 cm, quantitative MRI achieved an accuracy better than 95% in distinguishing between haemangiomas and malignant lesions. Routine quantitative measurements of relaxation times were sufficiently reproducible for clinical use. PMID- 2554384 TI - Microangiography and dynamic computed tomography in experimental chronic pancreatitis in the pig. AB - Chronic pancreatitis was induced in 22 piglets by dividing all pancreatic attachments to the duodenum. Ten piglets served as controls. The animals were subjected to dynamic computed tomography (CT) two, four or six weeks after operation. Microangiography was also performed on the pancreas two, four or six weeks after operation, following dynamic computed tomography. Microangiography revealed significant changes in the vasculature in advanced chronic pancreatitis. The arteries and arterioles were fewer than normal, their walls were thickened and their diameters exhibited variation. The capillary network was loose and the capillaries were not as well filled as in healthy animals. Contrast enhancement of the pancreas was, however, similar in all experimental animals to that in the healthy controls. It was concluded that the morphological changes seen in chronic pancreatitis histologically and on microangiography could not adversely affect contrast-enhanced CT because there were no changes in the function of the capillary network. On the basis of these results, dynamic CT should give no more information than CT with intravenous contrast medium in chronic pancreatitis. PMID- 2554385 TI - [Follow-up of portocaval anastomoses using DSA]. AB - In 37 patients with porto-caval shunts, the operation site was examined on 40 occasions by i.a. DSA (25 Warren shunts, four Linton shunts and eight Drapanas shunts). The problem of shunt persistence could be resolved on 39 occasions. Only once was it not possible to evaluate the shunt because of artifacts. Artifacts caused by breathing, peristalsis and low contrast in the portal-venous segment did not prove serious problems in evaluating shunt patency. Intra-arterial DSA provides a comprehensive demonstration of the portal venous vascular territory and permits evaluation of post-operative haemodynamics. PMID- 2554386 TI - [Transpenile venous occlusion in the treatment of erectile impotence]. AB - In 15 of 18 patients, 2 of them with previous surgery, transpenile venoablation was performed for treatment of erectile dysfunction due to cavernous leakage. In 4 this was combined with retrograde venooclusion via the internal iliac vein. In 4 patients normal erectile status could be obtained. In 3 other patients erections became possible with use of intracavernous vasoactive agents. In two only mild amelioration, and in two others no change of erectile dysfunction were obtained. Follow-up studies are yet missing in 3 patients. Complications of the procedure were not observed. PMID- 2554388 TI - [Digital multilayer tomography. A description of the method and initial clinical results]. AB - A set of projection images is acquired during longitudinal tomography with an image intensifier TV system. Reconstruction of tomograms in each desired plane is achieved by shifting and summing up of the digitalized projection images. Digital multilayer and conventional film tomograms mainly of the respiratory tract and skeleton have been compared in 100 patients. Image quality is comparable with both methods. Disadvantage of digital tomography is lower spatial resolution (512 x 512 matrix size); advantages include lower radiation dose, shorter study time, and facilities of digital imaging. PMID- 2554387 TI - [Analysis of dynamic CT curves in kidney transplants with rejection reaction and cyclosporin A poisoning]. AB - The early differential diagnosis between transplant rejection and cyclosporin intoxication is often difficult, although it is important for treatment. We carried out dynamic CT on 48 occasions in patients with normal function, with acute tubular necrosis, rejection and cyclosporin A intoxication. Analysis of the time-density curve of the cortex showed significantly slower enhancement with cyclosporin A intoxication than during rejection. The peak and the arterial curve were also delayed in cyclosporin A intoxication. This evidence of reduced cortical perfusion during cyclosporin A intoxication can be explained by the results of modern pathophysiological studies. In principle, it is possible to distinguish between cyclosporin A intoxication and rejection by means of dynamic CT, but for its routine use the nephrotoxicity of the contrast medium must be taken into account. PMID- 2554389 TI - [Measurement of noise levels in MR tomography at 1.5 Tesla]. AB - The noise levels at the isocentrum of an MR apparatus with a field strength of 1.5 Tesla was measured during various sequences. The rapid switching of the gradients exerts effects on the gradient coils, which result in the development of noise as from a loudspeaker. Measurements were carried out using gradients of 3 and 9.3 mT/m with head and body coils and a variety of sequences. The noise level depended largely on the type of sequence and, under extreme conditions, may attain 96 dB. Because of the short exposure times, we do not consider it necessary to use any form of protection. During the examination of patients with damaged hearing, some protection may be advisable. PMID- 2554390 TI - [The effective dose equivalent: clarification and comments on the new dosage concept]. AB - The concept of the effective dose, based on stochastic radiation effects (genetic damage and cancer), is explained and discussed. Reasons are given why this concept should consider the cancerogenic action of radiation only and not, in addition, the hereditary radiation effects. This view does not contradict our knowledge that the gonads have to be protected especially carefully from the mutagenic action of ionising radiation. PMID- 2554391 TI - Ileocolic intussusception due to an atypical Meckel's diverticulum. PMID- 2554392 TI - [Ileocolic invagination in Peutz-Jeghers syndrome]. PMID- 2554393 TI - [A massively dilated ductus choledochus in choledocholithiasis following cholecystectomy and choledochojejunostomy]. PMID- 2554394 TI - [A pseudotumor of the azygos septum]. PMID- 2554395 TI - [Systemic action of alcohol following sclerosis of hepatic cysts]. PMID- 2554396 TI - [Diagnosis of an angiosarcoma of the mammary glands]. PMID- 2554397 TI - [Multiple nodules in asymptomatic lobular panniculitis]. PMID- 2554398 TI - Osteoblastoma of the first metatarsal bone. PMID- 2554399 TI - [A chordoma located at L 1/2]. PMID- 2554400 TI - [Magnetic resonance tomographic detection of a primitive trigeminal artery]. PMID- 2554401 TI - [A comparison of the efficacy of sonography, computed tomography, ERCP and angiography in the diagnosis of primary bile duct carcinoma]. AB - Sonography, CT, ERCP and angiography were carried out in 22 patients with operatively and histologically confirmed primary carcinomas of the bile duct and the findings were compared with the appearances at operation. ERCP proved to have an accuracy of about 79% and is markedly superior to the other diagnostic methods. Sonography had an accuracy of about 39%, CT 42% and angiography 55%. PMID- 2554402 TI - [Acute intestinal hemorrhage in a pancreatic pseudocyst-colic fistula]. AB - Chronic pancreatitis with a pseudocyst may cause acute massive intestinal bleeding due to simultaneous erosion of the splenic artery and transverse colon. This is a potentially lethal complication, which requires instant diagnosis and treatment. The pathogenesis, diagnosis and treatment of this condition are illustrated by two patients. Exact localisation of the source of bleeding is achieved by selective angiography of the visceral arteries. In shocked and inoperable patients, selective embolisation of the bleeding artery is the treatment of choice. PMID- 2554403 TI - [Noninvasive diagnosis of splenic artery aneurysm]. AB - The findings and problems of differential diagnosis of splenic artery aneurysms are illustrated by three patients. By a combination of Doppler sonography and conventional duplex sonography, it was possible to achieve a correct and non invasive diagnosis. PMID- 2554404 TI - [Aggressive abdominal fibromatosis: the place of diagnostic imaging]. AB - Aggressive fibromatoses are locally invasive, non-metastasising, fibroblastic soft-tissue tumours. On the basis of examinations made in 6 patients with histologically confirmed diagnosis, the reliability of different imaging modalities in preoperative diagnosis and in follow-up is discussed. The inhomogeneous tumour composition was evident in all imaging procedures. Computed tomography offered the highest sensitivity for tumour detection and proved accurate in demonstrating the characteristic radial tumour spreading of mesenteric aggressive fibromatosis; the attenuation values ranged from 18 to 58 HU. in pre-contrast CT scans. In magnetic resonance imaging the tumours mostly produced a low signal in T1- and T2-weighted sequences; the calculated T2 relaxation times ranged from 97 to 186 ms. PMID- 2554405 TI - [Magnetic resonance tomography of children with surgically treated anal and rectal atresia]. AB - MRI of the pelvis was performed in 17 children following surgical correction of anal and rectal atresias and in five children without ano-rectal malformations. A muscle score was used to characterize the muscles of the pelvic floor and their relationship to the rectum. There was close agreement between the MRI muscle score and clinical continence. MRI provided additional information that should improve continence following conservative and surgical treatment. PMID- 2554406 TI - [Duplex sonography and magnetic resonance tomography in the clarification of nephrologic complications following kidney transplantation]. AB - A prospective study compared duplex sonography and magnetic resonance imaging in evaluating renal transplants. One hundred and two duplex sonographic and 24 MR examinations were performed and correlated with clinical course or biopsy. All normal renal allografts, 6 transplants with acute tubular necrosis and 2 cases of cyclosporin toxicity had normal Doppler waveforms, whereas 9 renal transplants with evidence of interstitial rejection by biopsy showed an obliteration or reversal of diastolic flow. MR imaging was less specific in identifying allograft rejection. There were false positive results in normal renal transplants, allografts with acute tubular necrosis and after rejection therapy. With regard to cost, accessibility and specificity, duplex sonography is the method of choice for the evaluation of renal allografts. PMID- 2554407 TI - [Obstetric pelvimetry using digital image intensifier radiography]. AB - Obstetric pelvimetry can be performed by digital image intensifier radiography using very low exposure doses. Comparative measurements show a reduction of the entrance dose to 5% of conventional respectively 15% of high speed film-screen radiography. Phantom measurements have shown an accuracy of +/- 5 mm. In 30 patients the transverse diameter of the pelvic entrance, of the interspinous and of the intertuberous level as well as the sagittal diameter of the pelvic entrance and outlet were measured. Pelvimetry is indicated for the early recognition of certain risks if anamnestic (previous Caesarean section), clinical (eg. external pelvimetry) or fetometric features indicate the evidence of disproportion. As a radiological method the image intensifier radiography appears particularly recommendable in respect of its low exposure dose and the possibility of interactive measurements with subsequent documentation of the diameters and values. PMID- 2554409 TI - [Intra-arterial DSA of the hand arteries in the diagnosis of inflammatory connective tissue diseases]. AB - Seventy-five patients with suspected rheumatic diseases underwent intraarterial DSA of the hand arteries. In 81% of the patients we were able to establish the diagnosis of vasculitis according to angiographic criteria. Neither clinical nor chemical or immunological features allowed the diagnosis of an inflammatory disease of joint and perivascular tissue at this time. However, histological examination and immunofluorescence microscopy confirmed the angiographic findings and showed signs of an immune complex vasculitis. A strong predictor for an abnormal angiogram of the hand arteries is the presence of Sicca's and Raynaud's Syndrome in addition to rheumatoid joint pain. PMID- 2554410 TI - [Diagnosis of deep venous thrombosis of the leg using color-coded duplex sonography]. AB - One hundred and twenty-nine lower limbs in 103 patients with suspected deep leg vein or pelvic vein thrombosis were examined by colour-coded duplex sonography and the results compared with phlebography. The deep veins could be demonstrated from the popliteal to the distal external iliac vein and internal structure and blood-flow could be evaluated. The procedure can diagnose not only occluding thrombus, but also thrombus with surrounding blood flow and post-thrombotic changes. Fresh venous thromboses can be recognised with a sensitivity of 96% and a specificity of 97%. Colour-coded duplex sonography is a simple, rapid and reliable method in all patients with suspected deep vein thrombosis. Our experience suggests that it can replace phlebography for the above-mentioned venous territories and problems. PMID- 2554408 TI - [Current indications for angiography of tumors of the muscular and skeletal systems]. AB - CT and MRI have greatly influenced the indications for angiography as a diagnostic method for tumours of the muscular and skeletal systems. 59 patients with bone and soft tissue tumours were examined by arteriography (DSA) and the indications for arteriography were compared with CT (32 cases) and MRI (15 cases). Particular attention was paid to the presence of vessel encasement. Arteriography is the definitive method for demonstrating vascular involvement. Tomographic methods demonstrate the topographic relationships of the tumour and neuro-vascular structures. Angiography can also be used as an interventional measure pre-operatively or for palliative embolisation. PMID- 2554411 TI - [Rotational angioplasty--recanalization of chronic arterial occlusion using a slowly rotating catheter. Clinical results with 100 patients]. AB - Between December 1986 and January 1989, 100 patients with chronic occlusions of peripheral arteries were treated with the new technique of low speed rotational angioplasty. This uses a relatively thick, flexible and blunt catheter, which is driven by an electric motor (100 to 200 r.p.m.). The success rate in the superficial femoral and popliteal arteries of the initial intervention for occlusions less than 10 cm was 90%, for occlusions of less than 10 cm, it was 80%. Occlusions on which conventional techniques had failed were successfully recanalized in 65%. Recanalisation was also successful in seven out of 12 patients with occluded iliac arteries. There were no perforations or other serious complications. It is concluded that the new method is particularly effective and safe for recanalisation of chronic vascular occlusions and should also be considered for cases which previously could only be treated surgically. PMID- 2554412 TI - [Limits of the diagnostic capability of pulmonary digital subtraction angiography]. AB - Digital subtraction pulmonary angiography was performed in 70 patients suspected of having pulmonary emboli. In 35 patients this diagnosis proved to be correct. The technical quality of all ECG-gated studies was scored and was found to be satisfactory in 31 patients (44%), moderate in 23 (33%) and unsatisfactory in 16 patients (23%). This was mainly due to the lack of patient cooperation. In patients with direct injection into the pulmonary artery (27%) the motion artifacts were less disturbing. We therefore suggest that even when using ECG gated acquisitions, the injection should be performed into the pulmonary artery to overcome technical problems in the diagnosis of pulmonary embolism. PMID- 2554413 TI - The radiology of the right superior intercostal vein. AB - The right superior intercostal vein is visualised on CT examination as a circular opacity laterally at the right aspect of the vertebral body at the T4-T5 level. In venographic examination the RSIV appears to be formed by the confluence of the venous channels, the right second, third and fourth intercostal veins. In superior vena cava obstruction the RSIV is an important collateral pathway: the flow through the vein depends on the site of the obstruction, anterograde if the obstruction is superior or parallel to the level of the vena azygos, and retrograde if the obstruction is below the vena azygos. PMID- 2554414 TI - [Percutaneous removal of a pacemaker electrode]. AB - The large number of pace-makers introduced transvenously has resulted in increasing the number of complications, amongst which infections and dislocations are prominent. It is usually necessary to remove the electrode, which can be attempted percutaneously by using a Dormia basket, a loop or forceps. Amongst the complications of this procedure are tears to the myocardium, with the risk of pericardial tamponade, or tears of the tricuspid valve leading to tricuspid insufficiency. Consequently, thoracic surgical intervention should be available if necessary. Four successful procedures are described. PMID- 2554415 TI - [Malignant chest wall infiltration in MR: comparison with CT and surgical findings]. AB - CT and MRI were performed on 19 patients with pleura related thoracic tumors with respect to the detection of chest-wall invasion. All patients underwent surgery, which confirmed malignant infiltration in 16 cases, while this was excluded in three. CT showed chest-wall invasion in 12/19 patients. MRI demonstrated tumorous involvement in 16 patients. A reliable pattern of chest-wall invasion in MRI were high signal intensity lesions within the chest-wall in the T2-weighted images. However, increased signal intensity of pleural structures was found in inflammatory as well as in malignant lesions. MRI can prove the presence of chest wall invasion when CT is equivocal. PMID- 2554416 TI - [Computed tomographic volumetry of the orbit in endocrine orbitopathy]. AB - The volumes of the four recti muscles and the orbital fat was measured by CT in 40 normal persons and in 60 patients with clinically confirmed Graves' disease. Compared with normal persons, 42 patients (70%) showed an increase in muscle volume and 28 patients (46.7%) an increase in the amount of fat. In nine patients (15%) muscle volume was normal, but the fat was increased. By using volumetric measurements, the amount of fat in the orbits in patients with Graves' disease could be determined. PMID- 2554417 TI - [MRT using gadolinium-DTPA in spinal intradural space-occupying lesions]. AB - The results of contrast enhanced MRI in 36 patients with suspected spinal intradural tumours are described. All intramedullary tumors showed distinctive enhancement and solid tumors could be delineated clearly, even if they were not clearly visible on unenhanced scans. The differentiation between neoplasm and non neoplastic syrinx was markedly improved. The sensitivity of MRI for demonstrating intradural extramedullary tumours was greatly improved by gadolinium DTPA and even small lesions or flat meningeal infiltrates could be visualised. In addition, gadolinium DTPA improved the delineation and localisation of larger lesions, even if they had already been seen on unenhanced images. PMID- 2554418 TI - [SPECT of the brain using 99mTc-HMPAO in patients with cerebrovascular disease: a comparison with CT. A report of 100 cases]. AB - In a retrospective study the sensitivity of SPECT and CT in detecting changes in cerebrovascular disease is compared, based on a group of one hundred patients. The higher sensitivity of SPECT with regard to right-positive results, especially in TIA and PRIND and the markedly lower number of false-negative results are underlined and discussed. The fact that both methods reveal only a slight sensitivity regarding the detection of changes in the basal regions of the brain is also pointed out, with SPECT proving to be the more sensitive of the two methods. PMID- 2554420 TI - [Symptomless spontaneous rupture of a pancreatic pseudocyst into the stomach]. PMID- 2554419 TI - [Erosive hemorrhage of the splenic artery in pancreatic pseudocyst]. PMID- 2554421 TI - Duodenal schwannoma. PMID- 2554422 TI - Sigmoido-rectal invagination secondary to carcinoma: findings on CT. PMID- 2554423 TI - Arteriosclerotic abdominal aortic aneurysm infected with Yersinia enterocolitica. PMID- 2554424 TI - [Unusual forms of retroperitoneal fibrosis (Ormond's disease)]. PMID- 2554425 TI - [A solid space-occupying lesion in the hilar region of the spleen as a rare site of a carcinoma of the pancreatic tail]. PMID- 2554426 TI - [Computed tomographic detection of cardiac luxation following left-sided extensive pneumonectomy]. PMID- 2554427 TI - Bilateral entrapment syndrome in lower extremities. PMID- 2554428 TI - [Intracerebral pneumatocele: CT and MR findings]. PMID- 2554430 TI - [A gluteal mass as the initial manifestation of a calcified hepatocarcinoma]. AB - A patient is reported who had a mass in the gluteal region of three years evolution that turned out to be a metastasis of hepatocarcinoma. The peculiarities of this case are commented, as well as the unusual form of presentation of the hepatocarcinoma. The prolonged clinical evolution and the presence of calcifications in the primitive tumor should be emphasized. PMID- 2554429 TI - [Inflammatory pseudotumor of the liver]. AB - Certain rare, reactive, non-neoplastic processes are defined under the term inflammatory pseudotumor, which is constituted by the proliferation of connective tissue and chronic inflammatory infiltrate of circumscribed location. Its etiopathogenesis ins unknown, although it is thought to have a possible infectious nature. It has been described various organs, but location in the liver is exceptional. We present a new case of inflammatory pseudotumor of hepatic location, related to an "hour-glass" abscess adhered to diaphragm, this being the first case of this relation. We reviewed the literature concerning etiopathogenic, histopathologic, clinical and diagnostic aspects, and our findings are discussed. PMID- 2554431 TI - [Hepato-splenic hemangiosarcoma: presentation of a clinical case]. AB - A 65-year-old farmer who had used arsenic as a plaguicide for many years developed a hepatosplenic hemangiosarcoma with metastasis in the colonic serosa, mesentery and omental. The tumor was complicated with intraabdominal hemorrhage originated by spontaneous intraperitoneal rupture. The echographic and post mortem findings are presented. This is the first case of hepatic hemangiosarcoma reported in Uruguay. PMID- 2554433 TI - The nerve growth factor receptor: biochemical and structural analysis. PMID- 2554432 TI - [Insulinoma. Apropos of a new case]. AB - Insulinomas are rare tumors whose difficult diagnosis is usually delayed with respect to appearance of the first symptoms. The Whipple triad (symptoms of hypoglycemia, low blood glucose and rapid alleviation of symptoms after glucose administration) associated with an increase in plasma immunoreactive insulin (IRI) and low blood glucose levels constitute the diagnostic basis of insulinoma as the cause of organic hypoglycemia. Curative treatment entails surgery and its success will depend on the location of the tumor. Selective arteriography and, more recently, percutaneous transhepatic insulin determination (IRI) in the splenoportal axis have a diagnostic rate of 90%. Nevertheless, none of these topographic diagnostic methods replaces meticulous intraoperative exploration of the pancreas, which in our case was definitive for cure. PMID- 2554434 TI - Regulation of steroid hydroxylase gene expression is multifactorial in nature. AB - In summary, regulation of steroid hydroxylase gene expression is complex and multifactorial, involving cAMP-dependent and -independent mechanisms required for maintenance of optimal steroidogenic capacity, tissue-specific mechanisms which lead to different steroidogenic pathways in different tissues, and developmental mechanisms which lead to fetal imprinting of steroid hydroxylase expression and which probably overlap with both maintenance and tissue-specific mechanisms. Future studies will involve identification of the trans-acting factors associated with each of these aspects of the multifactorial regulation and characterization of the cis-regulatory elements to which they bind. Such studies will inevitably lead to the identification of genes encoding these trans-acting factors and investigation of their regulation. In this way, it will be possible to work outward from the steroid hydroxylase genes toward the cell surface receptors in order to elucidate the series of events which lead to cAMP-dependent and independent regulation of steroid hydroxylase gene expression. PMID- 2554435 TI - Sequelae of SCI after discharge from the initial rehabilitation program. AB - The purpose of this study was to identify sequelae that occurred in persons with spinal cord injuries 11 weeks to 5 years after discharge from two initial rehabilitation programs. The investigator-developed instrument was first administered to a convenience sample of 21 subjects (Adamski, 1983). The modified version of the instrument was administered to a probability sample of 19 subjects in the present study. The results demonstrated that respondents experienced a higher incidence of medical sequelae than has been documented in the literature. These findings have important implications for nursing practice in developing and evaluating educational programs for persons with spinal cord injuries. PMID- 2554436 TI - The effects of boron hypolipidemic agents on LDL and HDL receptor binding and related enzyme activities of rat hepatocytes, aorta cells and human fibroblasts. AB - Two boron-containing hypolipidemic agents, N,N-dimethyl-n-octadecylamine borane and tetrakis-mu-(trimethylamine-boranecarboxylato)bis-(trimethylamine- carboxyborane)-dicopper(II) were shown to reduce significantly serum cholesterol and triglyceride levels after 14 days administration in rats. Serum low density lipoprotein (LDL) cholesterol content was reduced by both agents; however, high density lipoprotein (HDL) cholesterol was elevated by N,N-dimethyl-n octadecylamine borane but reduced by the dicopper (II) complex. The boron agents suppressed LDL binding and internalization in hepatocytes, fibroblasts and aorta cells and elevated HDL binding and degradation in hepatocytes. Enzyme activities were modulated by the boron derivatives which would favor less cholesterol deposition in peripheral tissues and further clearance from the plaque cells. PMID- 2554437 TI - Chrysotile asbestos-induced cytotoxicity and calcium-dependent exocytosis in polymorphonuclear leukocytes. AB - Chrysotile asbestos is cytotoxic for rabbit polymorphonuclear leukocytes (PMNs) which is evident from the release of the cytoplasmic enzyme LDH. In the presence of Ca2+, but not in its absence, a strong lysozyme release occurs. High concentrations of Ca2+ are required for exocytosis. In the presence of the stable guanine nucleotide GTP gamma S exocytosis occurs at lower Ca2+ concentrations. The results suggest that the plasma membrane is permeabilized by asbestos, allowing the influx of extracellular Ca2+ which causes exocytosis. At high Ca2+ concentrations asbestos-induced cytotoxicity is strongly reduced. The cytotoxic LDH release and, to a lesser degree, the exocytotic enzyme release are inhibited by Co2+, Mn2+ and La3+. Cytochalasin B and the glycolytic inhibitors 2 deoxyglucose and iodoacetate inhibit asbestos-induced cytotoxicity, indicating that microfilaments and glycolytic energy are required for the membrane-damaging actions of asbestos. PMID- 2554438 TI - Internalization of glucagon-like peptide-1(7-36)amide in rat insulinoma cells. AB - Glucagon-like peptide-1(7-36)amide [GLP-1(7-36)amide] is supposed to be an important physiologic incretin. Recently, high affinity receptors for GLP-1(7 36)amide have been demonstrated on rat insulinoma-derived RINm5F cells. The present study examined the internalization and degradation of the GLP-1-receptor complex. Internalization of the peptide was time- and temperature-dependent. At 37 degrees C binding and internalization was rapid. At 60 min 35% of 125I-labeled GLP-1(7-36)amide was internalized. Incubation in the presence of increasing concentrations of non-labeled GLP-1(7-36)amide resulted in a decrease of internalization of 125I-labeled peptide indicating that this process is saturable. Incubation in the presence of 0.2 mM chloroquine, an inhibitor of intracellular hormone degradation, resulted in intracellular accumulation of 125I GLP-1(7-36)amide. HPLC-supported analysis of cell content after internalization of 125I-GLP-1(7-36)amide during a 60-min incubation period at 37 degrees C revealed an elution profile showing two maxima of radioactivity: one represented intact labeled GLP-1(7-36)amide, the other an intracellular degradation product of the peptide. Chloroquine caused a 5-fold increase of the peak representing intact 125I-GLP-1(7-36)amide thus demonstrating inhibition of degradation of labelled peptide. Furthermore, a 4-fold increase of the other peak occurred possibly mirroring a delay of release of degradation products by chloroquine. It was excluded that chloroquine is able to interfere with GLP-1(7-36)amide-binding to its receptor. PMID- 2554439 TI - Relationship between pituitary ACTH content and hypothalamic catecholamines in the rat. AB - Hypothalamic concentrations of epinephrine and norepinephrine were determined in rats following 6-hydroxydopamine lesions of the locus coeruleus and subcoeruleus system and following sham-operation. These concentrations were correlated with pituitary ACTH content. While the lesion procedure did not have a major effect on hypothalamic monoamine levels, we were able to demonstrate a strong negative correlation between hypothalamic epinephrine and pituitary ACTH content independent of the experimental condition. Only a weak negative correlation was observed for hypothalamic norepinephrine and pituitary ACTH. Our recent and previous data suggest a tonic and phasic inhibition of ACTH release by hypothalamic monoamines. PMID- 2554441 TI - Re-evaluation of cardiopulmonary resuscitation. PMID- 2554440 TI - Inhibition of fog-induced bronchoconstriction by nedocromil sodium and sodium cromoglycate in intrinsic asthma: a double-blind, placebo-controlled study. AB - The efficacy of pretreatment with two doses of nedocromil sodium (4 and 8 mg) and sodium cromoglycate 12 mg were compared with placebo in inhibiting the bronchoconstriction induced by inhalation of ultrasonically nebulized distilled water ('fog') in 10 subjects with intrinsic asthma. Each fog challenge consisted of three inhalations of 30, 60 and 120 s duration, respectively, given at 4-min intervals, and the bronchoconstrictor response was assessed as the postchallenge percentage fall in FEV1 from baseline. Statistically significant drug effects (p less than 0.05) were observed after 120 s of fog challenge: both nedocromil sodium 4 and 8 mg and sodium cromoglycate were significantly more effective than placebo in inhibiting fog-induced bronchoconstriction; in addition, nedocromil sodium 4 mg proved statistically significantly more effective than sodium cromoglycate. PMID- 2554442 TI - Use of low-dose neostigmine intravenously in the treatment of supraventricular tachycardia: an immediate bradycardic effect. AB - Neostigmine was first used 50 years ago to treat sinus tachycardia and paroxysmal auricular tachycardia. Then there were reports of successful treatment by neostigmine of other forms of supraventricular tachycardias. However, reports of sudden death using neostigmine for reversal of neuromuscular blockade at the end of an operation, which were not properly treated with atropine abandoned the use of neostigmine as an antiarrhythmic drug. Low-dose neostigmine intravenously was used in the treatment of supraventricular tachycardia in three patients described herein. It gave an immediate bradycardic effect in all three patients. The use of a low-dose neostigmine intravenously for an immediate treatment of supraventricular tachycardia is a novel suggestion. It has to be further evaluated and compared to the conventional drugs used like digoxin, verapamil, propranolol or esmolol. PMID- 2554443 TI - Evaluation of cardiac vulnerability and antifibrillatory properties of anti arrhythmic drugs. AB - A method of evaluating the antifibrillatory properties of drugs by their effect on the acceleration of the cardiac rhythm by electric pulses was developed. It permitted measurement of fibrillation thresholds and the maximal driving frequency of stimulation. The heart was accelerated in closed chest dogs, and this increased the fibrillation thresholds after the application of lidocaine (1 mg/kg), quinidine (5 mg/kg), and novocainamide (15 mg/kg body wt.). The development of an original programmed stimulator increased the accuracy of the method by means of establishing the initial and terminal stimulation rates and observing the constant steps of change of pulse intervals. Four methods of causing fibrillation were compared: (1) a single pulse during the vulnerable phase of the cardiac cycle; (2) a train of pulses overlapping the vulnerable phase; (3) sequential R on T pacing; (4) simple acceleration of the cardiac rhythm. In addition to the other methods, the method of accelerating the heart rate differs in that only a small amplitude of stimulating pulses is needed. The present method may be used in the case of an unstable initial cardiac rhythm. PMID- 2554444 TI - The relationship of conjunctival and arterial blood gas oxygen measurements. AB - Conjunctival oxymetry (CjO2) measures peripheral tissue oxygen at the conjunctival level. CjO2 changes can indicate pulmonary or circulatory conditions leading to shock. Literature review does not define 'normal' CjO2/ABG PaO2 ratios. We designed a study to measure these ratios. Twenty-two healthy patients undergoing cardiac catheterization had simultaneous PcjO2 and PaO2 measurements completed. The range of conjunctival oxygen measurements was from 34 to 68 mmHg with a mean of 50.5 mmHg. The PaO2 readings ranged from 65 to 93 mmHg with a mean of 77.1 mmHg. The average PcjO2/PaO2 ratio was 0.656 with a range of 0.47-0.93. Thus the PcjO2 is on average 66% of the arterial blood gas PaO2. This ratio of 0.66 can serve as a base for further clinical studies in which PcjO2 is looked at in patients with pulmonary or circulatory illnesses or injuries. PMID- 2554445 TI - The effects of dihydroergocryptine on the neurological and enzyme disorders induced by cerebral ischaemia in rats. AB - Ergot alkaloids are commonly used as cerebroprotective drugs. Their efficacy has been demonstrated experimentally in animals submitted to acute cerebral anoxia or ischaemia, at dose levels hugely superior to dose levels usually administered in humans. In the present experiments, dihydroergocryptine (DHEC), a constituent of dihydroergotoxine (DHET), was administered at doses closely related to human doses, preventively (in experiments where animals survived only for a short while after ischaemic insult) or curatively, and its efficacy tested through refined neurological and biochemical evaluation of experimental cerebral ischaemia sequelae. DHEC was administered orally (30 micrograms or 150 micrograms/kg body weight (bwt) twice daily) for 3 days, following transient cerebral ischaemia induced by a 60-min carotid occlusion plus sodium nitroprusside (1.1 mg/rat s.c.) injection, or, in a second experiment, prophylactically (60 micrograms or 300 micrograms/kg bwt/day) for 4 days prior to multiple cerebral infarct induced by sodium arachidonate injection into the left internal carotid artery. The neurological sequelae were evaluated by the Irwin visual placing response or by a battery of behavioural tests. Na-K-ATPase enzyme activity in cerebral homogenates was measured; decreases in this enzyme activity are considered to reflect the neuronal membrane consequences of the neurocell energetic metabolism alterations caused by cerebral ischaemia. Low dose oral DHEC treatment prevented the behavioural abnormalities and memory impairment arising after transient cerebral ischaemia and there was a marked trend in improving the behavioural abnormalities observed in animals submitted to massive cerebral infarction, in spite of the model severity. DHEC prevented reduction in cerebral Na-K-ATPase activity after cerebral multiinfarction. These effects of DHEC were observed with doses and administration route close to the usual therapeutic regimen. PMID- 2554446 TI - The biological activity of adrenaline after injection through an intravenous cannula containing alkaline buffer. AB - Since it is known that alkaline pH inactivates adrenaline it has been recommended that this drug not be administered in the same i.v. line as alkaline buffer solutions during cardiopulmonary resuscitation. In order to test the validity of this statement a simulation model of the clinical situation was designed where the biological activity of adrenaline was measured in anesthetized rats after having been mixed with alkaline buffer solution contained in a standard i.v. cannula. The biological activity of adrenaline was measured by comparing the blood pressure response after repeated administration of a test (adrenaline + alkaline buffer) and control (adrenaline + normal saline) solution to a rat which had previously received a ganglion-blocking agent. Two alkaline buffer solutions, sodium bicarbonate and Tris buffer mixture were tested. These resulted in a decrease of the biological effect of adrenaline to 77 +/- 6 and 82 +/- 9% of control values, respectively. If however, adrenaline mixed with Tris buffer mixture was injected into a recipient of phosphate buffer (pH 7.40 and buffer capacity equal to human blood) as much as 94 +/- 17% of its activity remained. The results lead us to suggest that, in the cardiopulmonary resuscitation situation, adrenaline may well be given via a cannula containing alkaline buffer solution without significant interference with its effect. PMID- 2554447 TI - Plasma catecholamines, pH, and blood pressure during cardiac arrest in pigs. AB - This study examined plasma epinephrine (E) and norepinephrine (NE) concentrations, pH, and mean arterial blood pressure (MAP) in a cardiac arrest model. Twenty-three domestic swine (15-26 kg) were anesthetized with ketamine 20 mg/kg, i.m. and alpha-chloralose 25 mg/kg, i.v. and ventilated with a respirator. Catheters were placed in the right ventricle, left ventricle and femoral arteries for MAP recordings and blood pH sampling every 2 min. Catecholamine samples were taken from the femoral artery every 2 min. Cardiac arrest was induced by endocardial stimulation with a Grass S88 stimulator. Five minute post arrest resuscitation was initiated with a mechanical resuscitator. Ten minute post arrest NaHCO3 1 mEq/kg was administered by the peripheral i.v. (P; n = 6), central (CE; n = 5), or intraosseous, via the tibia, (I; n = 6), route. Controls (C; n = 6) did not receive NaHCO3. MAP (mean +/- S.D.) prior to arrest was: C 144 +/- 16, P 139 +/- 11, CE 137 +/- 13 and I 133 +/- 11 mmHg. Five and 25 min post arrest it was: C 21 +/- 5 and 17 +/- 6, P 34 +/- 8 and 23 +/- 7, CE 17 +/- 7 and 14 +/- 10 and I 26 +/- 6 and 11 +/- 3 mmHg, respectively. A 2-way analysis of variance did not reveal any difference in MAP values in the four groups. In all groups the blood pH from the femoral artery demonstrated a respiratory alkalosis that peaked at approximately 7.48 5 min after initiation of mechanical resuscitation. In the groups receiving NaHCO3, it peaked at 7.77 +/- 0.09 CE and 7.65 +/- 0.06 P 2 min post infusion and at 7.71 +/- 0.06 I 8 min post infusion. An analysis of variance revealed that the CE and I routes were significantly different (P less than 0.05) from the P group and that all three groups were different (P less than 0.05) from the C. Plasma E and NE concentrations at 0, 6, 10, 12, 20 and 30 min post arrest in the C group were, respectively: 3 and 10, 94 and 327, 119 and 329, 92 and 234, 33 and 135, and 127 and 62 ng/ml, respectively. All 3 groups receiving NaHCO3 demonstrated similar patterns and were not significantly different from C when compared with a 2-way analysis of variance.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2554448 TI - Initiation of closed-chest cardiopulmonary resuscitation basic life support. A personal history. AB - Modern external (closed-chest) cardiopulmonary resuscitation (CPR) basic life support (BLS) gives everyone anywhere a chance to initiate the reversal of death from airway obstruction, apnea, or pulselessness. The history of modern CPR has its roots around 1900, but lay dormant for half a century, until in the 1950s several fortunate circumstances merged to allow for documentation of Steps A (airway control by head-tilt and jaw-thrust), B (breathing control by mouth-to mouth ventilation), and C (circulation control by closed-chest cardiac massage, i.e. chest compressions) and their combination into BLS Steps A-B-C. BLS is only for borderline emergency oxygenation, i.e. Phase I of the life support chain. Both the non-authoritarian environment of the U.S.A. and several role players with keen interest in resuscitation were needed to enable the systematic research (Steps A and B), a chance rediscovery (Step C), and the integration of BLS with advanced life support (ALS, drugs and defibrillation, transferred from open-chest CPR) and brain-oriented prolonged life support (PLS, intensive care) to result in the development of an effective cardiopulmonary-cerebral resuscitation system. A fertile environment led rapidly to the development of resuscitation delivery systems in hospitals and communities. This paper is a story told by one of the role players. PMID- 2554449 TI - Results of cardiopulmonary resuscitation in a cardiology hospital. AB - The authors analysed a series of 557 consecutive patients who suffered cardiorespiratory arrest at the Dante Pazzanese Institute of Cardiology (DPIC) during a period of 5 years in order to examine factors predicting successful resuscitation and long-term survival. Cardiopulmonary resuscitation (CPR) maneuvers were tried in 536 patients, with the following results: 284 patients (53%) died immediately, another 102 (19%) died within the first 24 h after the cardiac arrest and 150 patients (28%) survived more than 24 h. Among these, 65 (12.1%) died in the first month after cardiac arrest and other 29 (5.4%) died after that period. There were 43 late survivors (8%). Thirteen patients (2.4%) were lost to follow-up. After 9 years, the accumulative life expectancy was 8.7%. Coronary heart disease, cardiomyopathy and valvular heart disease were the most frequent underlying diseases. None of the 49 patients with cyanotic congenital heart disease survived. The heart arrest was mostly caused by heart failure (55.8%) and primary arrhythmia (17.2%) in the whole group, whereas the survivor group showed primary arrhythmia in 81.7% and heart failure in 7.3%. In those patients where the initial mechanism of cardiac arrest was ventricular fibrillation, 33.2% survived more than 1 month, while among those on ventricular asystole, only 3.4% survived more than 1 month. PMID- 2554450 TI - Temporary organ substitution by hemoperfusion through suspension of active donor hepatocytes in a total complex of intensive therapy in patients with acute hepatic insufficiency. AB - The results of intensive therapy and temporary organ substitution by hemoperfusion through a suspension of active hepatocytes in 126 patients suffering from acute hepatic insufficiency (AHI) induced by virus B hepatitis, virus non-A, non-B hepatitis, acute toxic hepatitis, active liver cirrhosis, sepsis leptospirosis long-term subhepatic jaundice are presented in this paper. Hepatic encephalopathia confirmed both clinically and electroencephalographically was registered in all the patients. The patients were subdivided into two groups: a complex of commonly used curative measures according to the intensive therapy for AHI was applied in Group A (67 patients); in Group B (59 patients), alongside with the above measures, temporary organ substitution by hemoperfusion through a suspension of active porky hepatocytes was also performed. The lethality in Group A made up 59% and that in Group B was 37%. PMID- 2554451 TI - Esophageal tracheal combitube (ETC) for emergency intubation: anatomical evaluation of ETC placement by radiography. AB - The esophageal tracheal combitube (ETC) is designed for emergency intubation. The ETC is inserted blindly allowing ventilation after either esophageal or endotracheal placement. A special pharyngeal balloon serves to seal the upper airways. In 10 cardiac arrest patients, emergency intubation with the ETC was performed. Blood gas analyses showed adequate ventilation. Radiography proved correct placement of the proximal and distal balloons in accordance with design theory. Hyperinflation experiments documented expansion of the proximal balloon into the oral cavity rather than towards the epiglottis. PMID- 2554452 TI - Solute back-diffusion raises the gas concentrating efficiency in counter-current flow. AB - We have extended the counter-current model of the rete mirabile of the fish swimbladder to include the effects of inert gas secretion into the swimbladder as well as solute back-diffusion in the rete capillaries. (1) Gas secretion attenuates the inert gas concentrating efficiency of the rete, i.e. its ability to produce high inert gas partial pressures in blood at the swimbladder pole. The maximum attainable gas secretion rate depends on the salting-out effect, i.e. on the ratio of solubility in venous and arterial blood of the rete. (2) Solute back diffusion leads to a significant increase in the concentrating efficiency, and this is due to the salting-out effect produced by the solute when it diffuses back into the arterial capillary blood. This enhancement is particularly prominent when gas and solute permeabilities of the rete vessels are high. (3) Estimates for physiologic parameters in the European eel suggest that lactate back-diffusion may contribute significantly to the gas concentrating efficiency of the rete mirabile. PMID- 2554453 TI - A new in vitro screening system for anticancer drugs for the treatment of non small cell lung cancer. AB - We have evaluated a semiautomated radiometric assay (BACTEC 460 system) for screening of activity of anticancer drugs against human non-small cell lung cancer cell lines. Cells from seven cell lines were exposed to standard antineoplastic agents at four different concentrations using a 1-h incubation. Alpha 2-interferon was tested using a continuous incubation. In vitro drug activity was analyzed as a function of the clinically achievable serum concentration. Our results indicate that two cell lines (CALU-3, SK-MES-1) exhibit in vitro drug sensitivity patterns closest to those observed in clinical studies. These two cell lines might therefore be most useful for screening new anticancer compounds for activity against non-small cell lung cancer. The radiometric assay is a semiautomated system which has advantages over other, more time-consuming screening systems. PMID- 2554454 TI - Infection prevention and control in the era of the AIDS/HIV epidemic. AB - The chronology of important events in the AIDS/HIV epidemic is presented in Table 2. Nurses are the largest group of health care workers and persons who provide direct care and handle sharp objects regularly. As the number of persons infected with HIV increases and as more infected persons become ill enough to require care, the opportunity for an individual nurse to have contact with an infected person will increase. Because many persons with HIV infection also have Kaposi's sarcoma and other malignancies, some oncology nurses may have more contact with HIV-infected individuals than nurses in general. However, it is well established that the major risk for HIV infection is from puncture injuries, and a number of strategies are available and are being developed to make needle and sharps handling safer. Nurses must take personal responsibility for knowing how to reduce their own risks for exposure to HIV and other infectious agents while keeping in mind the need not to increase risks of nosocomial infections in patients. Thus, all nurses need a comprehensive understanding of the purposes of barriers (eg, gloves, gowns, masks, handwashing, room assignment) and when and how to use them correctly. In addition, all nurses who have contact with blood and body fluids should take responsibility for their own immunity to hepatitis B by obtaining hepatitis B vaccination. By using all of these strategies in combination, many infection risks to nurses and patients will be minimized. PMID- 2554455 TI - [Variations in plasma angiotensin-converting enzyme and renin in sarcoidosis]. AB - A comparative study of plasma angiotensin-converting enzyme and renin activity was carried out in 55 patients with sarcoidosis and other pulmonary diseases. The converting enzyme activity was significantly increased in active sarcoidosis, showing a tendency to return to normal values after corticoid treatment, while plasma renin activity remained unaltered both before and after corticotherapy. The obtained results confirm the usefulness of the converting enzyme measurements, as already shown by others. Attention is drawn on the possible involvement of the pulmonary renin-angiotensin system in the local vascular and enzymo-chemical alterations, suggesting the possibility of associating the converting enzyme inhibitors to the corticotherapy of sarcoidosis. PMID- 2554457 TI - Long-term tissue distribution and steady state activity ratios of 232Th and its daughters in rats after intravascular injection of Thorotrast. AB - To estimate the absorbed dose in the critical organs of Thorotrast patients, it is necessary to know not only the distribution and concentration of 232Th but also its daughter nuclides in the body. The present investigation was undertaken in order to clarify the long-term 232Th tissue distribution and steady state activity ratios between subsequent daughters in the critical tissues using about 30 Wister male rats, as a basis for estimating absorbed doses. The tissue distribution of thorium was examined by means of an autoradiography of the whole body and/or the gamma-ray spectrometry at various times during 2 to 24 months following injection. The concentrations of daughter nuclides in tissues were determined by repetitive gamma examination over a period from 1 hr to 35 days after being sacrificed. The data indicate (1) that approximately 90% of injected Thorotrast is retained in the body for a prolonged period, but about 50% of radium and 10% of radon produced from thorium are eliminated from the body, (2) that the mean steady state activity ratios of 224Ra and 212Pb to 228Th for liver are 0.56 and 0.28, and 0.54 and 0.16 for spleen, 0.58 and 0.82 for lungs, respectively, and (3) that the parent 228Th is translocated to the bone. PMID- 2554456 TI - [The involvement of adenosine and adenosine deaminase in experimental myocardial infarct]. AB - By the ligature of the left coronary artery in the rat anesthetized with nembutal (10 mg/100 i.p.) a significant increase of the 5'-nucleotidase activity (Wooton method) was noticed 10 minutes after the left ventricle infarction (from an average value of 1038.5 +/- 187 mU/g tissue to 1537 +/- 225 mU/g fresh tissue). The adenosine desaminase levels spectrophotometrically determined by Denstedt technique, do not appear significantly modified 10 or 30 minutes after the left ventricle infarction. The chromatographically determined adenosine levels, by HPLC technique, decrease from the average value of 11.63 +/- 1.4 micrograms/mg PT to 8.60 +/- 1.0 micrograms/mg PT 30 minutes after infarction. The observed changes are explained by the conditions of hypoxia in the infarcted ventricle which lead to the raise in adenosine levels by activating the 5'-nucleotidase and their depression by a very fast metabolism of the same substance. PMID- 2554458 TI - [Clinical experiences of LAK therapy in patients with hepatocellular carcinoma]. AB - A patient with hepatocellular carcinoma (HCC) was treated with newly established adoptive immunotherapy using LAK cells (LAK therapy) together with transcatheter arterial embolization therapy (TAE). This patient responded well, and the therapeutic efficacy still continues 6 months after the therapy. Since the efficacy of LAK therapy does not last long, it is recommended that LAK therapy should be employed in combination with such therapeutic maneuvers as TAE or anticancer drugs in patients with HCC. PMID- 2554459 TI - [A case from practice (151). Metastasizing, hepatocellular carcinoma with small nodular, probably alcoholic liver cirrhosis]. PMID- 2554460 TI - Fixation and orientation control by the tecto-reticulo-spinal system in the cat whose head is unrestrained. AB - The role of the tecto-reticular and tecto-reticulo-spinal neurons (here called TR(S)Ns) in gaze control is described. TR(S)Ns, located in the deeper layers of the cat superior colliculus (SC), project onto the eye and head premotor circuitry. TR(S)Ns located in the caudal SC had sustained and phasic discharges related to the control of gaze movements. The sustained discharge occurred when the visual axis was positioned at some vector quantity away from a target of interest. Each cell has its preferred vector corresponding to the cell's location on the collicular retinotopic map. This tonic discharge acted as a preamble to the phasic discharge and served to pre-excite the relevant oculomotor circuitry. The phasic discharge preceded gaze shifts whose direction and magnitude matched the preferred vector. The intensity of this discharge was correlated to the acceleration and velocity of the movement. TR(S)Ns situated in the rostral SC were maximally active when the cat fixated a target of interest. These neurons decreased their discharge rate during gaze shifts. Thus, TR(S)Ns provide both fixation and orientation signals to the eye and head premotor circuitry. A scheme is proposed where TR(S)Ns lie within a gaze feedback loop that controls eye and head movements via inputs to long lead burst neurons and omnipause neurons. PMID- 2554461 TI - [Amebiasis. Its epidemiology today]. PMID- 2554462 TI - [Amebiasis. Features of its immunologic behavior]. PMID- 2554463 TI - [Amebiasis. Has its clinical expression changed?]. AB - In Mexico, like in other countries, there is an special interest for amebiasis because it represents a Public health problem; there are about 5 million people that in some time of their lives had tissue invasion by this parasite. 1-2% of mexicanas have intestinal amebiasis and probably there are 850,000 to 1,700,000 carriers of entamoeba cysts. 6% of the general population have circulating antiameba antibodies. In 1988 we studied the amebic hepatic abscess seen in the hospitals of the Instituto Mexicano del Seguro Social in the Valley of Mexico. During that year there were 396 patients with such illness (0.19% of hospital admissions) with a 1.01% mortality. Comparing this data with reports of 1969 there was a reduction in the number of patients and deaths. Also amebiasis has decreased in autopsy studies. However, we have not found recent variations in the clinical behavior of amebiasis. Advances in diagnostic methods and more liberal use of metronidazole have reduced the number of severely ill patients. PMID- 2554464 TI - [Amebiasis. The technology of the 80's. II. Studies of fecal material and serology for the diagnosis of amebiasis: have they helped improve the diagnosis of hepatic abscess?]. PMID- 2554465 TI - [Combatting smoking--the primary measure for preventing pulmonary diseases]. PMID- 2554466 TI - [Rare syndromes in pulmonary pathology]. PMID- 2554467 TI - [Factors in the failure of tuberculosis chemotherapy studied in a Romanian county]. AB - This work proposes to analyse the quality of the failure risks in the case of a short duration treatment applied in operational conditions on the territory of Constanta County as a premise for the application of a method of mathematical analysis on computer in view of predicting the failure cases. There were studied 262 eligible cases out of 290, which represent the total of the patients newly registered with pulmonary tuberculosis in 1982, on the territory of Constanta County and who were kept an eye upon for 3 years after finishing the treatment. There resulted 58 (22.1%) recurrencies, failures or corrected in course intratherapeutic positive reactions, on which were analysed 25 possible failure factors. The most representative have proved to be the organizational factors among which irregular treatment (67.2%) of the failure cases, a defective mobilizing to treatment, technical factors: late negativation of sputum (over 4-5 months from starting the treatment), and among the factors depending on the patients--noncooperation occurred in 60.33% of the failure cases. If the final frequency of failures on the territory of Constanta County does not exceed the average figures recorded here and abroad, the global frequency within the studied time period is not satisfactory and imposes an assessing of the failure factors and measures of improvement. PMID- 2554468 TI - [Pleural puncture biopsy--a diagnostic method in serofibrinous tuberculous pleurisy in children]. AB - The biopsy of the parietal biopsy with the help of a needle has become an essential method of diagnosis in the adult patients with a pleural malady of an unknown etiology and may become a complementary paraclinical method in children. A sure diagnosis in the tubercular serofibrinous pleurisy with children is made by rendering evident the Koch bacillus on the direct test or in culture and by rendering evident the tubercular lesions on fragments of pleural biopsy puncture. The study is supported on the results obtained at the pleural biopsy puncture performed on a number of 6 children. In four cases out of six the histological test of the pleural fragment has rendered evident the presence of tubercular lymphoepithelioid nodulus with central necrosis, thus carrying the argument of certitude. The pleural biopsy puncture allows an early and sure diagnosis in over two-thirds of the pleurisies of a tubercular etiology. PMID- 2554469 TI - Pulmonary mechanics in patients with diffuse interstitial lung disease and normal lung volume. AB - Static expiratory pressure-volume curves obtained in 20 patients with diffuse interstitial lung disease and lung volume within normal ranges (i.e. in patients in an early stage of the disease) showed different patterns of abnormal lung elastic recoil: 1) P-V curves with normal shape but shifted to the right, which suggest an overdistension of a reduced number of functioning alveoli ("lung shrinkage") and 2) flattened P-V curves (i.e. with low static compliance) also shifted to the right, which argue for an abnormal distensibility of most alveoli or a more diffuse spreading of lesions. These patterns suggest a multifactorial genesis of the increased elastic recoil in diffuse interstitial lung disease. PMID- 2554470 TI - Growth of M. tuberculosis H37Rv strain in 7H12 Middlebrook liquid medium in the presence of acridine orange and THA. AB - The influence of acridine orange and THA on the growth of Mycobacterium tuberculosis H37Rv strain in 7H12 Middlebrook liquid medium was studied. As compared to acridine orange, THA also inhibited the growth of the H37Rv Mycobacterium tuberculosis strain but at much higher concentrations. PMID- 2554471 TI - [Characteristics of the evolution of pulmonary tuberculosis in a female patient with Wegener's granulomatosis]. PMID- 2554472 TI - [Characteristics of the onset of pulmonary cancer in children and adolescents]. PMID- 2554473 TI - [The advantages and limits of computed tomography (compared with conventional methods) in the diagnosis of pulmonary adenocarcinoma (a case report)]. AB - The authors analyse the advantages offered by the computerized tomography in comparison with the conventional methods of diagnosis, the study and classification (TNM) of a case of pulmonary adenocarcinoma with multiple metastases: renal, suprarenal, hepatic, peritoneal, bony, lymph node. At the same time there are also pointed out the limits of this new method. PMID- 2554474 TI - [The present situation of osteoarticular tuberculosis in the general endemic context of tuberculosis in the Socialist Republic of Romania]. PMID- 2554475 TI - [Secondary prevention in chronic obstructive bronchopneumopathies]. PMID- 2554476 TI - [The medical and social-occupational aspects of the sequelae of osteoarticular tuberculosis]. PMID- 2554477 TI - [Recent occurrence of human infection by Rocio arbovirus in the Valley of Ribeira region]. AB - The presence of IgM antibodies to Rocio in sera of two children from rural area of Ribeira Valley, Brazil, was detected by MAC-ELISA. This new arbovirus of the Flaviviridae family was responsible for an extensive encephalitis epidemic that occurred in the region in 1975-1977. Since 1980 no human disease caused by this virus has been diagnosed. An improvement on surveillance of Rocio infections and on the researches for virus identification in suspected vectors and reservoirs is necessary. PMID- 2554479 TI - Carcinoembryonic antigen, arginine vasopressin and calcitonin as markers of early small-cell lung cancer relapse. AB - Patients with small-cell lung cancer (SCLC) underwent serial blood monitoring during remission: 66 were tested for carcinoembryonic antigen (CEA) and calcitonin (CTN), including 40 who were concomitantly tested for arginine vasopression (AVP) as well. 83% of the patients had at least one assay elevated prior to induction therapy. By serial monitoring, blood concentrations reflected disease course and reached lowest levels at remission. CEA, AVP and CTN shifted along the course of disease independently of each other; a normal pretreatment titer did not preclude its rise at a later phase, while an initially elevated assay could normalize at remission and stay normal thereafter. The median lead time (LT) to clinically diagnosed relapse, for limited-disease patients, was 229 days for complete and 90 days for partial remitters. Patients with extensive disease had similar LT values. LT to local recurrence was shorter than to distant relapse. Remission AVP titers of up to 6 ng/ml conferred disease-free survival (DFS) longer than that associated with higher titers (median DFS 518 vs. 211 days, respectively; p = 0.045 for curve differences). The relative risk (RR) of relapse associated with pretreatment patient characteristics and with absence or presence of tumor marker normalization at remission was estimated by the Cox proportional hazards model. This analysis revealed that the RR of relapse conferred by pretreatment attributes, e.g. disease extent, was considerably modified by biochemical co-variates at remission, e.g. serum CEA level relative to a 3 ng/ml cutoff point. PMID- 2554478 TI - Iron resistance of hepatic lesions and nephroblastoma in rainbow trout (Salmo gairdneri) exposed to MNNG. AB - Histochemical markers are important for the early detection of chemically initiated neoplasia in experimental animal studies. The marker, iron resistance, was evaluated in the Shasta strain of rainbow trout (Salmo gairdneri). Twenty-one day-old trout embryos were exposed to 100 ppm aqueous N-methyl-N'-nitro-N nitrosoguanidine (MNNG) for 30 min in a static water bath. Fish were fed a semipurified diet, and sampled monthly from the 4th to the 9th month. Two days before sampling, fish were iron-loaded with a single ip dose of 0.30 mg iron dextran/100 g body weight. Livers and kidneys were conventionally processed to paraffin sections for iron, or hematoxylin and eosin (H&E) staining. Normal hepatocytes accumulated iron in pericanalicular locations, but in hepatocytes from carcinogen-altered foci and tumors, iron staining was clearly reduced or absent. Normal renal tubule cells exhibited slight to moderate iron staining, while those from nephroblastoma were iron resistant. These results establish iron resistance as a property of preneoplastic and neoplastic trout hepatocytes and nephroblastoma cells for the first time. Iron resistance may offer a practical histochemical marker in experimental fish models of hepatocellular carcinoma and nephroblastoma. PMID- 2554480 TI - Application of monoclonal antibody KH2 against lactosyl and neolactotetraosyl ceramide to immunohistochemical study of human cancers. AB - Monoclonal antibody KH2 (IgM) reactive with lactosyl ceramide and neolactotetraosyl ceramide was applied for the first time to the staining of human cancerous and noncancerous tissues which were fixed with formalin and embedded in paraffin. Digestive-tract cancers originating from stomach, colon, pancreas, gallbladder and bile duct were clearly stained. The staining intensity was especially strong in the apical portion and the intraluminal content of adenocarcinomas. In noncancerous tissues, it was of interest that gastric intestinal metaplasia and certain fetal tissues were stained. A part of the tubules of the kidney, pancreatic ductal cells, lung alveolar epithelial cells and polymorphonuclear leukocytes were stained in normal adult tissues in our study. Enhancement of the intensity of immunostaining by neuraminidase treatment indicated that most of the antigenic determinants should be sialylated and the antigenicity exposed after neuraminidase treatment in several kinds of cancerous and noncancerous tissues. These data suggest that the monoclonal antibody KH2 may be useful in the immunohistologic diagnosis of formalin-fixed human cancerous tissues. PMID- 2554481 TI - [Treatment of mycoses and new antifungal agents]. AB - The multiplication of iatrogenic factors, nosocomial diseases and acquired immunodeficiency syndrome is responsible for an ever increasing number of deep opportunistic mycoses, namely candidiasis, aspergillosis and cryptococcosis. The advent of a wide variety of rare opportunistic fungi is a fairly recent and worrying phenomenon. To combat these infections, often very serious, our therapeutic armentarium is rather scanty. Moreover, the prescription of the available drugs is limited by their toxicity, their spectrum and their route of administration or by the emergence under treatment of resistant mutants. Beside old products, such as amphotericin B and 5-fluorocytosine, miconazole and, mainly ketoconazole (both azole derivatives) were the first steps towards oral administration and low toxicity. Fluconazole is a triazole antifungal compound with a very original distribution to the meninges and urinary tract; it is mainly used in candidiasis and cryptococcosis. Another triazole compound, itraconazole, presents the particularity of being active against Aspergillus spp., Cryptococcus spp. and some agents of exotic mycoses. These two products are well tolerated and should soon be available for use in deep visceral mycoses. Other azole derivatives are under study. Among compounds issued from new chemical families, terbinafine (allylamine) is particularly active against dermatophytes, and cilofungine (a polypeptide) against fungi. These drugs are in the experimental stage. Research should be pursued aimed at developing, probably in new chemical families, and agent that is fungicidal in vivo. PMID- 2554482 TI - [Human herpesvirus 6 and AIDS]. PMID- 2554483 TI - [Transfusion and transmission of infectious and parasitic diseases excluding AIDS and viral hepatitis]. AB - Many viruses, bacteria or parasites can survive in stored blood for varying lengths of time. Recipients are therefore exposed to a risk which depends on the prevalence of pathogens in blood donor populations, the clinical and laboratory controls performed in blood transfusion centres and the efficiency of the patient's immune system. Beside the HIV and hepatitis viruses, transfusions may transmit the HTL virus in endemic areas or if the blood donor comes from one of these areas (e.g. the French West Indies), the CMV virus (but only in patients with weak immune defences) and some exotic viruses in specific regions. As regards bacterial agents, syphilis is prevented by blood storage at 4 degrees C for 72 hours and brucellosis remains a minor risk, but the very rare endotoxinic shock is severe and lethal in two-thirds of the cases. Infestation by parasites is common in certain areas, but it may occur in France after transfusion from blood donors coming from these areas; malaria transmitted by blood perfusion is a real problem. Drastic procedures of rejection of blood donors at risk, including examination and laboratory screening, must be applied and are effective in preventing these dangers. These procedures are well-known and are compulsory in France. PMID- 2554484 TI - [Heparin and its low molecular weight derivatives]. PMID- 2554485 TI - [Erythroblastopenia in children]. AB - The occurrence of isolated erythroblastopenia is a common problem in paediatrics, and in most cases three diagnoses may be considered: Blackfan-Diamond anaemia (congenital erythroblastopenia), transient erythroblastopenia of childhood and erythroblastopenia consecutive to parvovirus B19 infection. These three diseases have distinctive features: age of onset, association with other laboratory and clinical abnormalities and above all, transient or chronic character are of assistance in making an exact diagnosis. The mechanisms responsible for Blackfan Diamond anaemia and transient erythroblastopenia are imperfectly known. In vitro studies of the properties of erythroblast progenitors suggest intrinsic damage to these cells, whereas in transient erythroblastopenia their differentiation seems to be inhibited, perhaps by an immune mechanism. In both cases, the abnormality has not be identified. The selective tropism of parvovirus B19 towards actively dividing erythroblasts accounts for the acute erythroblastopenia observed in a context of chronic haemolysis. The responsibility of parvovirus B19 in chronic or relapsing erythroblastopenia in immunocompromised patients is a recent discovery. In view of the protean haematological pathology in this context and of the difficult therapeutic problems it creates, this diagnosis must systematically be envisaged, notably in children under chemotherapy. PMID- 2554486 TI - Ginkgo biloba extract prevents mucosal damage associated with small-intestinal ischaemia. AB - We have examined how a Ginkgo biloba extract influences the damaging effects of ischaemia in the small-intestinal mucosa. We used a rat experimental model in which a ligated loop of the distal ileum was subjected to ischaemia and revascularization, and the ensuing mucosal damage assessed by lysosomal enzyme release and intestinal permeability measurements. We also determined the mucosal content of malondialdehyde, a lipid peroxidation product, and the mucosal activity of myeloperoxidase, a neutrophil granulocyte marker. Ischaemia and revascularization alone caused increased mucosal permeability to sodium fluorescein, increased N-acetyl-beta-glucosaminidase release from the mucosa into the lumen, increased malondialdehyde content in the mucosa, and increased myeloperoxidase activity in the mucosa. Intravenous injection of G. biloba extract caused a dose-dependent attenuation of all these effects of ischaemia. It is suggested, therefore, that G. biloba extract may protect the intestinal mucosa against ischaemic damage by reducing neutrophil infiltration and lipid peroxidation. PMID- 2554487 TI - Interaction of rioprostil with the parietal cell prostaglandin E2 receptor. AB - The interaction of rioprostil with the porcine parietal cell prostaglandin (PG) E2 receptor and with uterine 3H-PGE2 binding sites was investigated. The in vitro antisecretory effect was measured by the inhibition of histamine stimulated 14C aminopyrine uptake in isolated pig parietal cells. Rioprostil displaced 3H-PGE2 competitively from the porcine fundic PGE2 receptor and had 1/25 of the affinity (Kd = 8 X 10(-8) mol/l) of the natural ligand; it was, however, equipotent with the synthetic prostacyclin analogue, iloprost. Similar affinity ratios were found in uterine muscle. Rioprostil inhibits parietal cell acid production with an IC50 in the range of its gastric binding dissociation constant. It is concluded that rioprostil exerts its antisecretory effects via a parietal cell E-type receptor. The compound cannot discriminate between fundic and uterine 3H-PGE2 binding sites with as yet unknown functional implications. PMID- 2554488 TI - Enhancement of human immunodeficiency virus (HIV) replication in human monocytes by low titres of anti-HIV antibodies in vitro. AB - Low concentrations of serum obtained from a patient with acquired immunodeficiency syndrome (AIDS) enhanced the replication of human immunodeficiency virus type 1 (HIV-1) in a particular subclone of the CD-4 positive monocytoid cell line U937 clone 2. Cells of this subclone have a high expression of Fc receptors and a considerable degree of Fc-mediated phagocytic activity. IgG purified from the serum was also able to enhance the replication. These results indicate that low concentrations of human anti-HIV antibody may enhance HIV replication on human monocyte-macrophages. Furthermore, two mouse IgG1 monoclonal antibodies against gp120, the envelope glycoprotein of HIV-1, also induced enhancement at low concentrations. The binding of radiolabelled gp120 to the cells was increased at the same low concentrations. Antibodies against envelope glycoproteins may cause enhancement of HIV infection. Both normal and enhanced replication of HIV were completely inhibited by the masking of the binding site of CD4 molecules with F(ab')2 fragments of anti-CD4 antibody. Moreover, CD4-positive, Fc gamma RI-negative K562 cells and mouse macrophages failed to show any infection in the presence of antibody. These results suggest that CD4 molecules on the cell surface are necessary to cause enhancement of infection of HIV on monocyte-macrophages. PMID- 2554489 TI - Anti-idiotypic B-cell lines from a patient with monoclonal gammopathy of undetermined significance. AB - The B-cell repertoire in a patient with benign monoclonal gammopathy of unknown significance was studied using Epstein-Barr virus transformation of peripheral lymphocytes. The presence of anti-idiotypic B cells producing monoclonal antibodies that reacted with idiotypic determinants on the monoclonal immunoglobulin was verified. The two monoclonal anti-idiotypic antibodies studied were of the IgM-kappa type. Such anti-idiotypic antibodies may be part of an idiotypic network regulation of the monoclonal B-cell population. PMID- 2554491 TI - [Neurotoxicity with cytostatic therapy: a review]. AB - Neurotoxicity is of increasing importance in the treatment of cancer patients. Since the dose-limiting hematological toxicity of most cytostatic drugs can now be overcome by modern supportive therapy, high-dose chemotherapy with curative intent can be used more often. Non-hematological toxicities such as neurotoxicity will therefore become dose-limiting with greater frequency, and to avoid disabling neurological side effects of chemotherapy the drug-induced neurological disturbances must be recognized before irreversible damage has occurred. The neurological side effects of the cytostatic drugs are reviewed. PMID- 2554490 TI - Gamma-interferon-mediated down-regulation of electrolyte secretion by intestinal epithelial cells: a local immune mechanism? AB - Active chloride (Cl-) secretion by intestinal crypt enterocytes is the central pathophysiological disturbance in most cases of acute diarrhoea. We examined monolayers of the human intestinal cell line T84 mounted in Ussing chambers to see whether the T-cell lymphokine gamma interferon (IFN-gamma) might affect the Cl- secretory properties of these cells, which morphologically and functionally resemble native crypt enterocytes. Pretreatment of T84 cell layers with IFN-gamma for 24 h (but not for 3 h) markedly decreased the Cl- secretory response to vaso active intestinal polypeptide (VIP) and to cholera toxin and carbachol without appreciably affecting the overall morphology, electrical resistance, or cyclic AMP response of the T84 cell monolayer. The IFN-gamma treatment, however, did induce subtle changes in the T84 cell membrane protein composition which might have affected ion channels regulating Cl- secretion. Our results may indicate a possible novel 'cell-mediated' immune mechanism through which activated gut T cells could modulate the extent of intestinal electrolyte and fluid secretion in, for example, enteric infections. PMID- 2554492 TI - Stone to bone. PMID- 2554493 TI - Synapse formation in the developing brain. PMID- 2554494 TI - p53: a frequent target for genetic abnormalities in lung cancer. AB - Allele loss is a hallmark of chromosome regions harboring recessive oncogenes. Lung cancer frequently demonstrates loss of heterozygosity on 17p. Recent evidence suggests that the p53 gene located on 17p13 has many features of such an antioncogene. The p53 gene was frequently mutated or inactivated in all types of human lung cancer. The genetic abnormalities of p53 include gross changes such as homozygous deletions and abnormally sized messenger RNAs along with a variety of point or small mutations, which map to the p53 open reading frame and change amino acid sequence in a region highly conserved between mouse and man. In addition, very low or absent expression of p53 messenger RNA in lung cancer cell lines compared to normal lung was seen. These findings, coupled with the previous demonstration of 17p allele loss in lung cancer, strongly implicate p53 as an anti-oncogene whose disruption is involved in the pathogenesis of human lung cancer. PMID- 2554495 TI - A liver-specific enhancer in the core promoter region of human hepatitis B virus. AB - An 88-base pair fragment in the core promoter of the human hepatitis B virus (HBV) contains a functional promoter and a strong liver-specific enhancer. This enhancer functions in human hepatoma cells, where it is much more active than the previously described HBV enhancer in stimulating expression of the linked bacterial chloramphenicol acetyltransferase gene expressed from heterologous promoters. Studies of the role of this enhancer-promoter in HBV may help to clarify mechanisms of gene expression in cells infected with HBV and the role of the virus in the pathogenesis of hepatitis and hepatocellular carcinoma. PMID- 2554496 TI - Germ-line transmission of a c-abl mutation produced by targeted gene disruption in ES cells. AB - A substitution mutation has been introduced into the c-abl locus of murine embryonic stem cells by homologous recombination between exogenously added DNA and the endogenous gene, and these cells have been used to generate chimeric mice. It is shown that the c-abl mutation was transmitted to progeny by several male chimeras. This work demonstrates the feasibility of germ-line transmission of a mutation introduced into a nonselectable autosomal gene by homologous recombination. PMID- 2554498 TI - In vitro protamine neutralization profiles of heparins differing in source and molecular weight. PMID- 2554497 TI - Laboratory control of heparin and heparin fragment treatment: an experimental study. PMID- 2554499 TI - Potentiation by heparin fragment CY 222 (Choay) of thrombolysis induced by human tissue-type plasminogen activator. PMID- 2554500 TI - Strategy for the development of the low molecular weight heparin fraction CY 216 in the prevention of postoperative deep vein thromboses in general surgery. AB - The polymorphism in the composition and actions of heparin and the parallel complexity of the thrombotic process have led to the design of a cautious, methodical, and pragmatic program of development of the first low molecular heparin fraction presently available in France (CY 216). The main idea was always to present the objective observation of facts and to reject any theoretical construction. After many stages, the value of this fraction in this indication, which appears as safe as possible but more effective than unfractionated heparin, has become obvious. PMID- 2554501 TI - Clinical potential of low molecular weight heparins. PMID- 2554502 TI - Comparative human pharmacology of low molecular weight heparins. AB - The pharmacodynamics of LMW heparins differ consistently from those of normal heparin. The relative bioavailability is greater than 90% and for some LMW heparins more than 100% after subcutaneous administration, as measured by the anti-Factor Xa activity. This indicates release of endothelial bound glycosaminoglycans with anti-Factor Xa-like activities. Direct determination of the LPL and HTGL activities demonstrate higher capacities of two and a lower capacity of three LMW heparins to release these enzymes from their endothelial glycoprotein receptors into the bloodstream. Thus, all LMW heparins are characterized by improved pharmacodynamics compared with normal heparin, but their anti-Xa to APTT and anti-Xa to anti-IIa ratios and half-lives differ among each other. The neutralization of the anticoagulant effects of LMW heparins by protamine are similar in vitro and in vivo at equigravimetric doses. However, unfractionated heparin and LMW heparins are not completely antagonized by protamine on a whole blood clotting test (thrombelastography). This inhibition of Factor Xa of LMW heparins is counteracted not completely by protamine. This may be due to a release of other glucosaminoglycans by LMW heparins. The present overview demonstrates that a comparative human pharmacology gives useful devices for prophylactic and therapeutic regimen for LMW heparins in patients in addition to the data obtained from animal models. PMID- 2554503 TI - An objective evaluation of the clinical potential of low molecular weight heparins in the prevention of thromboembolism. PMID- 2554504 TI - Fibrinolytic and thrombolytic parameters in patients with deep vein thrombosis treated by low molecular weight and standard heparin. PMID- 2554505 TI - Chemical and biological heterogeneity in low molecular weight heparins: implications for clinical use and standardization. PMID- 2554506 TI - Oxidative stress: a radical view. PMID- 2554507 TI - Postoperative care after liver transplantation. AB - The proliferation of liver transplant services across the country during the next 5 years will be an interesting phenomenon to observe. An enlightened approach to the process require answers to several essential questions, many of which are discussed elsewhere in this issue of Seminars. Regardless of how the future distribution of services sorts out, many physicians will face an increasing likelihood that their services will be requested in the long-term management of liver transplant recipients. In this report, the care of liver transplant patients is discussed, including that involving the immediate post-operative phase, the in-hospital recovery, and the early and long-term care of outpatients. The discussion is limited to the more common problems that these patients experience. Information from available literature is interspersed with our experience to provide a personal approach to the diagnosis and treatment of both complications and normal post-transplant clinical concerns. Part of what the reader should obtain from a general review of a complex subject is an awareness of the amount of information that the review does not contain. We have touched the surface of a large number of topics, but have come up far short of a comprehensive coverage of all aspects of the postoperative care of the liver recipient. It is hoped that we have provided enough information to serve the reader as an initial foundation for building a more complete understanding of the management of liver transplant patients. PMID- 2554508 TI - [Treatment of severe cytomegalovirus infections with DHPG]. PMID- 2554509 TI - [Hormone treatments of advanced breast cancers]. PMID- 2554510 TI - Suboptimal effect of daptomycin in the treatment of bacteremias. AB - We have reported two cases involving bacteremic patients who failed to respond adequately to the investigational agent daptomycin. Despite apparent sensitivity of the organisms, therapy was unsuccessful in both patients using the recommended dosage. The sponsor of daptomycin is currently reevaluating the recommendation and may be revising their dosage guidelines in the future. PMID- 2554511 TI - Small cell carcinoma of the pleura. AB - Extrapulmonary small cell carcinoma is uncommon. Small cell carcinoma of pleural primary origin has not, based on a literature review of the past three decades, been reported. We have reported a case of small cell carcinoma of the pleura, discussed its management and significance, and briefly reviewed the pertinent literature. PMID- 2554512 TI - Severe pancreatitis in an AIDS patient in association with cytomegalovirus infection. AB - We have reported a case of pancreatitis caused by CMV in an AIDS patient. Although CMV has caused clinical disease of other gastrointestinal organs, pancreatic involvement has previously been demonstrated only in autopsy specimens. The etiology of "idiopathic" pancreatitis in AIDS patients should be investigated, as therapeutic intervention may be available for some of these persons. PMID- 2554513 TI - Small cell lung cancer in a patient with neutropenia due to Felty's syndrome. AB - A patient with severe neutropenia due to Felty's syndrome had limited stage small cell lung cancer, resulting in a therapeutic dilemma. Because he had a cellular bone marrow and no history of recurrent infection, we decided to administer full doses of cytotoxic chemotherapy with curative intent. A durable remission was achieved without complication, despite persistent severe peripheral neutropenia. It may be possible to administer aggressive chemotherapeutic regimens to selected patients with Felty's syndrome when there is the potential for cure of malignant disease. PMID- 2554514 TI - Technetium-pertechnetate uptake by ectopic antral mucosa. An experimental study. AB - The ability of ectopically placed fundic mucosa and retained antral mucosa to concentrate technetium has previously been demonstrated. The ability of ectopically placed antral mucosa to concentrate technetium was investigated. The denervated antrum implanted into the colon concentrated technetium in 5 dogs. However, this response was significantly less than that of parietal cell containing mucosa (P less than 0.005) of the stomach. Technetium scanning may be useful in detecting ectopic and retained antral mucosa. PMID- 2554515 TI - National surveillance for neonatal herpes simplex virus infections. AB - To explore the magnitude and descriptive epidemiology of neonatal herpes simplex virus (HSV) infection, the Centers for Disease Control (CDC) conducted an 18 month hospital-based surveillance study in which 184 cases were reported. Only 22% of mothers had a history of genital HSV infection and only 9% had genital lesions at the time of delivery. Cesarean delivery initiated prior to membrane rupture failed to prevent infection in 15 cases. These data confirm previous observations that most mothers of infected neonates have no history of genital HSV and are asymptomatic at delivery. Furthermore this study suggests that intrauterine infection may be an important route of transmission and underscores the limitations of current prevention strategy. PMID- 2554516 TI - Condom use and use-effectiveness in high-risk populations. AB - In vitro evidence strongly suggests that latex condoms provide an impenetrable barrier for the passage of infectious agents, but few have studied the use effectiveness of condoms in preventing sexually transmitted diseases. In 1971, a prospective evaluation of the condom failed to demonstrate efficacy against infections with gonorrhea because users were exposed to infectious secretions before the condom was used. An ongoing multicenter study of female prostitutes in the United States shows that none of 22 women who always use condoms with all male partners has antibody to human immunodeficiency virus (HIV), but differences in HIV antibody status by patterns of condom use are not statistically significant. An educational campaign to promote the proper and consistent use of condoms as prophylactics is needed to stop the spread of gonorrhea, infections with HIV, and other sexually transmitted diseases. PMID- 2554517 TI - Management of massive ovarian tumors. AB - Two patients with massive ovarian tumors, one with a 51 kilogram Stage IC mucinous cystadenocarcinoma and the other with a 34 kilogram mucinous cystadenoma, are presented. Problems associated with resection of massive ovarian tumors, including respiratory failure, intraoperative fluid shifts, adequate exposure, orthostatic hypotension and adynamic intestine, are identified. Guidelines for avoiding these pitfalls by the use of appropriate monitoring, controlled drainage of the cyst and transverse elliptic incision with abdominoplasty are suggested. PMID- 2554518 TI - The use of lasers in the treatment of cholangiocarcinoma. AB - Six of seven patients were treated with external beam radiation, iridium-192 internal radiation and neodymium-YAG laser therapy via a percutaneous transhepatic tract for relief of biliary obstruction. After laser therapy, the transhepatic catheters were removed and patients remained tube-free and non jaundiced for periods ranging from 1 to 20 months. PMID- 2554519 TI - Prospective comparison of preoperative imaging and intraoperative ultrasonography in the detection of liver tumors. AB - A prospective evaluation of the accuracy of preoperative computed tomography (CT), ultrasonography (US), and angiography was performed in 54 patients undergoing resection of hepatic neoplasms. The results were compared with surgical findings and intraoperative ultrasonography (IOUS). A total of 167 lesions was seen by means of IOUS, of which preoperative US enabled detection of 127 (76%). In 48 patients CT allowed detection of 91 of 150 lesions (61%), and in 35 patients angiography showed 56 of 107 lesions (52%). When the detection rate is analyzed according to hepatic segment, the greater overall accuracy of preoperative US may be attributed to a markedly better detection rate in lateral segment of the left lobe of the liver. Lesion size also represented a factor, with preoperative US allowing detection of a greater number of small (less than 2 cm) lesions compared with CT. In patients studied with both CT and US, the combined lesion-detection rate increased to 81% in the right lobe and 76% in the left lobe. Because of this we recommend that preoperative assessment include both CT and US evaluation of the liver. IOUS showed 25% to 35% additional lesions compared with preoperative US and CT. More importantly, 40% of the lesions demonstrated by IOUS were neither visible nor palpable at surgery. We recommend that IOUS be considered in all patients in whom resection of hepatic neoplasm is planned. PMID- 2554520 TI - Biologicals, standards and heparin. PMID- 2554521 TI - A second plasma inhibitor of activated protein C: alpha 1-antitrypsin. AB - Inactivation of activated protein C (APC) in normal human plasma was studied in the absence and presence of heparin. In the absence of heparin APC inactivation followed pseudo-first order kinetics. In the presence of heparin the neutralization of APC was found to be biphasic. Up to 500 nM APC could be readily inactivated in normal plasma, indicating that the concentration of the APC inhibitor must be higher than previously assumed. Plasma deficient in the protein C inhibitor (PCI-I, as described by Suzuki and coworkers) and deficient in beta 2 glycoprotein I still possessed APC neutralizing capacity, presumably through the formation of complexes of APC with another plasma protein as was demonstrated by immunoblotting with anti-protein C antibodies. Together these data made us to conclude that a second inhibitor of APC (PCI-II) must be present in normal human plasma. This second inhibitor should be heparin independent, have a relatively high plasma concentration and form complexes with APC. Subsequently, we purified this PCI-II by isolating APC-PCI-II complexes from plasma deficient of vitamin K dependent proteins, PCI-I and beta 2-glycoprotein-I, to which purified human APC had been added. Purified PCI-II has a molecular weight of 50,000 daltons and aminoacid analysis revealed that PCI-II is identical with alpha 1-antitrypsin (alpha 1-AT). The second order rate constant for the reaction between purified alpha 1-AT and APC was found to be 269 M-1 min-1 in the absence of calcium and 602 M-1 min-1 in the presence of calcium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554522 TI - Antithrombin III and hepatocellular carcinoma. PMID- 2554523 TI - Could thrombus age be a modulator of heparin or L.M.W.H. activities? AB - The main thrombotic diseases are caused by old constituted thrombi. However, experiments to demonstrate the effects of heparin or heparin fragments on tPA release have been on fresh thrombi. This study on thrombi induced 6, 24, 48 or 72 hours before sampling shows variations in the main biological activities of both heparin and heparin fragment (CY222) as the thrombus ages. This effect is particularly observed on tPA release which is statistically reduced (p less than 0.001). Thrombus age seems to be a modulator of heparin and heparin fragment biological activities. PMID- 2554525 TI - Beneficial effect of low molecular weight heparin on the hemodialysis model in dogs. AB - The effect of FR-860, one of low molecular weight heparins, was investigated on the hemodialysis model in dogs for comparison with that of conventional unfractionated heparin (UF-heparin). In a bolus injection model, FR-860 at 12.5 100 U/kg prolonged the dialysis time (the time until the arterial circuit pressure reaching 500 mmHg) in a dose-dependent manner and UF-heparin at 50 U/kg also prolonged it. Additionally, FR-860 (12.5-50 U/kg/hr) and UF-heparin (25 and 50 U/kg/hr) continuously inhibited the rise of the arterial circuit pressure in an infusion model. In both models, the efficacy of FR-860 was more potent than that of UF-heparin. FR-860 and UF-heparin decreased the amount of blood loss remaining in the dialyzer in the infusion model. During hemodialysis, both FR-860 and UF-heparin showed activated plasma anti-F.Xa activity, prolongation of aPTT and such of thrombin time in a dose-dependent manner in those models. However, FR 860 was higher in anti-F.Xa activity and weaker in prolongation of aPTT and thrombin time than UF-heparin. These results suggest that FR-860 is more beneficial in in efficacy and higher in safety against the bleeding risk than UF heparin hemodialysis. PMID- 2554524 TI - Comparative pharmacology of low molecular weight heparins: implications of manufacturing processes on biological effects. AB - With the recent development of numerous low molecular weight heparins (LMWHs), a certain amount of concern has become evident as to the equivalency of each agent. In a comprehensive study, we have taken the seven available LMWHs to directly compare their in vitro and in vivo (subcutaneous) antithrombotic properties in one laboratory setting. Where possible, various batches of one LMWH were evaluated. Our findings were that variations of in vivo activity were observed between the LMWHs studied. Some activities were significantly different from placebo, whereas others were not. Depending on the assay chosen significant differences could also be observed for the in vitro activity. PMID- 2554526 TI - Sheep platelets as a model for human platelets: evidence for specific PAF (platelet activating factor) receptors. AB - In a number of animals, the platelet response to Platelet Activating Factor (PAF) has been shown to differ considerably from that in humans. However, aggregation, release and particularly shape change were quite similar for human and sheep platelets. In this study, aggregation and shape change analysis were used to measure the response of sheep platelets to various synthetic analogs of PAF. Response is greatly reduced with no alkyl group in position 2 of PAF and decreases progressively as the number of carbons of the alkyl gets larger than three. A reduction of activity is also seen as the ether linkage at position 1 of PAF is replaced by an ester linkage. These changes are indicative of a specific membrane receptor for PAF in sheep platelets and confirm the usefulness of sheep platelets as a model for PAF-platelet interaction in humans. PMID- 2554527 TI - [Investigation of Cushing's syndrome. The diagnostic value of the dexamethasone suppression test, the metopirone test and the CRF test]. AB - We describe thirty-one patients with Cushing's syndrome, with the object of evaluating the relative merit of the Dexamethasone suppression test, Metyrapone test and Corticotrophin Releasing Factor (CRF) test in classifying the syndrome. Bilateral adrenocortical hyperplasia (Cushing's disease) was present in sixteen patients. Three had bilateral macrodular hyperplasia of the adrenal cortex, six had adrenocortical adenoma, four had adrenocortical carcinoma, and two patients presented ectopic ACTH-syndrome. The diagnosis was surgically verified in every case. The Metyrapone test was found to give the safest classification in patients with Cushing's syndrome. The Dexamethasone test will diagnose Mb. Cushing reliably when suppression of serum cortisol is present following the large dose of Dexamethasone, but failure to suppress does not exclude the diagnosis. The CRF test is easy to perform and distinguished reliably between Mb. Cushing and other causes of the syndrome in eight out of ten patients in whom it was performed. Outpatient examination including the CRF test and CT-scanning of the pituitary and adrenal glands is advocated as a preliminary step in the classification of biochemically and clinically suspected cases of Cushing's syndrome. PMID- 2554528 TI - [Perinatal mortality in calves with symptoms of weak calf syndrome]. AB - Seventeen calves died about the time of birth on a cattle farm within five months. The clinical findings were in accordance with the clinical symptoms of the weak calf syndrome. The BVD virus was identified in blood taken prior to the colostrum in one calf. PMID- 2554529 TI - [Oxygen radicals: rescue or threat?]. AB - In this review the chemistry and formation of oxygen radicals is surveyed. Various physiological protective mechanisms against oxygen radicals as well as the consequence of too much or too little oxygen radicals in diseases is discussed. In this respect attention is given to hypoxia/ischemia, bronchopulmonary dysplasia, retrolental fibroplasia, thalassaemia, Down's syndrome and chronic granulomatous disease. Finally some new developments in pharmacotherapeutic possibilities which might play a role in prevention or amelioration of free radical pathologies are mentioned. PMID- 2554530 TI - [Electron microscopic detection of herpesviruses in a mass death of Greek tortoises (Testudo hermanni) and four-toed turtles (Agrionemys horsfieldii)]. AB - In a population of about 130 tortoises, consisting mainly of Hermann's tortoises (Testudo hermanni) and Four-toed tortoises (Agrionemys horsfieldii), 50% of the reptiles died within 3 months in autumn 1987. Post mortem analysis of 5 necropsied turtles revealed thick, caseous coatings in the upper digestive tract, hepatomegaly and enteritis. By microbiological examination a lot of ubiquitous bacteria could be isolated. By electron microscopy of concentrated and negatively stained gut specimens herpes virus particles were found in two Hermann's tortoises and one Four-toed tortoise. Isolation experiments in embryonic turtle tissue cultures showed no evidence of a cytopathogenic agent. The results are discussed in respect of their importance for the high mortality observed in the population of mediterranean land turtles. PMID- 2554531 TI - Culture and changes in dynamics of proliferation of pulmonary small cell carcinoma cells. AB - We had developed a method for the selective cultivation of cancer cells in short term which was simple and required minimal technical skills. There have been so far 23 cases of pulmonary small cell carcinoma in which long-term passage of more than one year and establishment of a cell line were successful by the culture method. In particular, from November 1982 to March 1987, there were 29 patients who were diagnosed as having pulmonary small cell carcinoma on preoperative cytodiagnosis and whose cells were primary cultured. In 24 (83%) of these, isolation could be carried out in a short period and stable passage was possible. Long-term passage of more than one year and establishment of a cell line were successful in 21 (72%) of the 29 patients. As a result of primary culture of small cell carcinoma cells, a pattern of a suspended colony was observed in 22 (88%) of 25 cases. As passage advanced, the tendency toward attachment to the basal surface increased. The changes in dynamics of proliferation after long-term passage of more than one year were divided into three types: epithelial like cell type was observed in 8 (32%), the neurite like cell type was observed in 4 (16%) and the suspended colony type was observed in 13 (52%) of the 25 cases. Variant clones composed of relatively large cells appeared as a result of long-term passage or the use of carcinostatics in some pulmonary small cell carcinoma cells. PMID- 2554532 TI - Evidence for a role of tert-butyl hydroxylation in the induction of pneumotoxicity in mice by butylated hydroxytoluene. AB - Previous studies have shown that BHT must be biotransformed, probably to a quinone methide, in order to cause pneumotoxicity in mice. When BHT is incubated with mouse hepatic or pulmonary microsomes, a major metabolite that is formed is the tert-butyl-hydroxylated derivative of BHT (BHT-BuOH). Herein we show that BHT BuOH has a fourfold greater potency than BHT in increasing the lung wt/body wt ratio, decreases lung cytosolic Ca2+-dependent protease activity at 1/10 the dose required for BHT to do this, and causes pulmonary histopathology at 1/20 the dose of BHT. Lung damage occurs earlier and is repaired faster at lower concentrations of BHT-BuOH than of BHT, but the nature of the damage (type I cell death) and regenerative response (type II cell hyperplasia and differentiation) is apparently identical. Neither BHT-BuOH nor BHT cause damage to liver, kidney, or heart as assessed by light microscopy, so they are both specific pulmonary toxicants. We postulate that BHT-BuOH formation is an essential step in the conversion of BHT to the ultimate pneumotoxin, which might be the corresponding quinone methide. PMID- 2554533 TI - A comparison of three nucleoside analogs with anti-retroviral activity on immune and hematopoietic functions in mice: in vitro toxicity to precursor cells and microstromal environment. AB - A number of 2'3'-dideoxynucleosides have been reported to markedly inhibit the in vitro growth of HIV, the causative agent of acquired immunodeficiency syndrome (AIDS). Clinical trials have shown that the continued therapeutic use of these nucleoside derivatives can be associated with adverse side effects. Since these side effects include myelotoxicity, as occurs in many patients treated with zidovudine (AZT; 3'-azido'3-deoxythymidine), and AIDS patients already represent an immunologically compromised population, we examined the immunological effects of three nucleoside inhibitors, including zidovudine, 2'3'-dideoxycytidine, and 2'3'-dideoxyadenosine (DDA) in a mouse model. Additional studies were conducted to further determine and characterize the potential toxic effects associated with these drugs on the hematopoietic system. Of the three dideoxynucleosides examined, only DDA altered immune functions following a 30-day subchronic exposure in mice. This was evidenced by a marked suppression of the antibody plaque-forming cell response and a slight alteration in macrophage function. None of the nucleoside derivatives affected bone marrow function following in vivo exposure, although AZT produced a mild macrocytic anemia in vivo and was myelotoxic when added in vitro to bone marrow cell cultures. In vitro studies indicated that AZT was capable of affecting both proliferating stem cells as well as the stromal cell microenvironment, both of which play a role in hematopoiesis. These data indicate that, although the mice may not develop the identical toxicities associated with nucleoside therapy in humans, certain adverse immunological and hematological effects were readily discerned which could have relevance to humans. PMID- 2554535 TI - A possible cellular mechanism of cisplatin-induced nephrotoxicity. AB - Cisplatin, a relatively new antitumor agent, is associated with renal function impairment. The mechanism of cisplatin-induced nephrotoxicity is unknown. A mouse model was used to examine nephrotoxicity induced by cisplatin. This study demonstrates both morphologically and biochemically that mitochondrial damage may be associated with cisplatin-induced cellular toxicity. The morphological changes are evident after 72 h following a single 10 mg/kg i.p. dose of cisplatin. Biochemical changes also follow the morphological abbreviations. In vitro incubation of cisplatin with cells also shows a decline in Rhodamine 123 fluorescence with time, which is indicative of mitochondrial damage. The present findings suggest the possibility that the nephrotoxic effects of cisplatin may be related to a mitochondrial damage. PMID- 2554534 TI - Factors influencing the induction of DNA single strand breaks in rats by 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD). AB - The ability of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) to induce lipid peroxidation and DNA damage in rat hepatic nuclei was investigated. In addition, the role of iron in hepatic DNA damage was examined. Female Sprague-Dawley rats were treated with a single dose of 25--100 micrograms TCDD/kg orally, and sacrificed 3-14 days after treatment. Liver nuclei were isolated, and DNA single strand breaks (DNA-SSB) and lipid peroxidation were determined. Lipid peroxidation was assessed by measuring the content and production of thiobarbituric acid reactive substances (TBARS) while DNA-SSB were determined by the alkaline elution technique. The results demonstrate that TCDD dose and time dependent increases in hepatic nuclear TBARS content and production occur in conjunction with an increase in DNA-SSB. The administration of the dithiolthione antioxidant oltipraz (30 mg/kg/day for 10 days) resulted in a significant decrease (47%) in the incidence of TCDD-induced DNA-SSB. Clofibrate administration (100 mg/kg/day for 12 days) and pair feeding had no effect on TCDD induced DNA-SSB. The incubation of hepatic microsomes and mitochondria from TCDD treated rats with nuclei from untreated animals for one hr at 37 degrees C resulted in enhanced DNA damage which was abolished by the addition of 0.10 mM desferrioxamine (DFX). Incubation with cytosol had no significant effect. Incubation of 0.10 mM Fe2+ or Fe3+ with isolated hepatic nuclei from untreated rats produced significant increases in DNA-SSB, which were abolished by the addition of 0.10 mM DFX to the incubation medium. TCDD may produce an increased bioavailability of iron which leads to enhanced DNA single strand breaks and lipid peroxidation in hepatic nuclei. PMID- 2554536 TI - Interaction of Clostridium perfringens delta toxin with erythrocyte and liposome membranes and relation with the specific binding to the ganglioside GM2. AB - The specific interaction of the cytolytic Clostridium perfringens delta toxin with membrane GM2 was indicated by: (i) characterization of this glycolipid in the membrane of sheep and goat erythrocytes, which are lysed by the toxin, whereas GM2 was undetectable in insensitive rabbit erythrocytes, (ii) demonstration of 125I-toxin binding to GM2, by autoradiography, following incubation with thin-layer chromatograms containing separated neuroblastoma gangliosides, and (iii) toxin fixation by phospholipid-cholesterol unilamellar vesicles containing either sheep gangliosides or GM2. In order to investigate the intramembrane events leading to membrane disruption following toxin binding, the photoreactive probe 12(4-azido-2-nitrophenoxy)stearoyl 1-14C glucosamine, which inserts into the outer layer and labels integral membrane proteins, was used to establish whether delta toxin penetrates into target cell membrane. No toxin labeling was found, suggesting that toxin action takes place at the membrane surface. This contention is supported by the observation that despite toxin binding, GM2 liposomes did not release entrapped 14C-glucose. Treatment of toxin with carboxypeptidases, but not aminopeptidases, abolished both toxin binding capacity onto erythrocytes and its combination with antitoxin neutralizing antibodies, suggesting that the carboxy terminal end of the toxin is critical for binding to cell membrane. PMID- 2554538 TI - Influence of trichloroethylene treatment on phosphoinositides in rat brain. AB - The effect of oral administration of trichloroethylene, a neurotoxic solvent, on the levels of phosphoinositides in rat brain was studied. Two hours after administration of a single dose of trichloroethylene (1000 mg/kg body wt.), the levels of phosphatidylinositol (PI) and phosphatidylinositol 4,5-biphosphate (PIP2) were reduced by 24 and 17%, respectively, without any significant change in that of phosphatidylinositol-4-phosphate (PIP). Twenty hours after treatment, the levels of PI and PIP2 were increased by 22 and 38%, respectively. Repeated administration of the same dose of trichloroethylene for 1 year markedly reduced the levels of PI (52%), PIP (23%) and PIP2 (45%). These results for the first time suggest the involvement of a phosphoinositide messenger system in trichloroethylene neurotoxicity. PMID- 2554537 TI - Decreased phosphofructokinase activity during the development of triorthocresyl phosphate-induced delayed neuropathy. AB - The effect of triorthocresyl phosphate on selected glycolytic enzymes (hexokinase, phosphofructokinase, glyceraldehyde 3-phosphate dehydrogenase, lactate dehydrogenase) was investigated during the development of organophosphate induced delayed neuropathy. Only phosphofructokinase activity was decreased 15 days after treatment. The effect was dose-dependent and was observed in sciatic nerve while the brain enzyme activity was not affected. PMID- 2554539 TI - Acetone compared to other ketones in modifying the hepatotoxicity of inhaled 1,2 dichlorobenzene in rats and mice. AB - The ability of acetone and 3 other ketone vapours to influence the hepatotoxicity of inhaled 1,2-dichlorobenzene (DCB) was examined in rats and mice. Methylethylketone, methylisobutylketone or cyclohexanone increased liver cytochrome P-450 content and glutathione-S-transferase (GST) activity, but did not affect serum glutamate dehydrogenase (GLDH) activity in rats. Pre-exposure to these ketones enhanced DCB-induced increase in serum GLDH activity (8-63-fold), while the increases in cytochrome P-450 content (33-86%) and GST activity (42 64%) were identical to those resulting from exposure to ketones alone. Each of the 3 levels of exposure to acetone elicited cytochrome P-450 and GST responses comparable with those caused by the other ketones. In spite of that, acetone pre exposure potentiated (4785 ppm), reduced (10670 ppm) or suppressed (14790 ppm) DCB-induced liver toxicity. In mice, the 3 ketones mentioned above interacted with DCB on centrolobular liver glucose-6-phosphatase (G-6-Pase) while acetone pre-exposure elicited an interactive G-6-Pase response in the mediolobular area alone, suggesting topographic change. PMID- 2554540 TI - Increased availability of mercury in rat hepatocytes by complex formation with diethyldithiocarbamate. AB - The availability of different chemical forms of mercury (Hg) was studied in primary cultures of rat hepatocytes incubated with mercuric acetate (HgAc), mercuric diethyldithiocarbamate (Hg(DTC)2) or methyl mercuric chloride (MeHgCl), labelled with 203Hg. The uptake of Hg was linearly related to the concentration in the medium and increased in the order Hg(DTC)2 greater than MeHgCl greater than HgAc when similar concentrations of Hg were used. A maximum concentration of Hg was reached after 4 h incubation with Hg(DTC)2 while incubation with HgAc and MeHgCl resulted in a slow continuous accumulation of Hg for up to 24 h. Hg added as HgAc was bound to proteins in the incubation medium to a greater extent than Hg added as Hg(DTC)2 or MeHgCl. Differences in affinity to the medium, as well as in lipophilicity, may partly explain the observed differential uptake of these Hg compounds. The enzyme alcohol dehydrogenase was inhibited by HgAc and Hg(DTC)2 to a similar extent at comparable cellular concentrations of Hg. On the other hand, glutathione reductase was inhibited to a higher degree by HgAc than by Hg(DTC)2, indicating that Hg(DTC)2 remains at least temporarily in the complexed form and that the enzyme is less susceptible to Hg in this form. Both enzymes were much less susceptible to MeHgCl than to HgAc or Hg(DTC)2. The results from the present study indicate that diethyldithiocarbamate can increase the transport of Hg across the cellular membrane by complex formation with Hg and thereby increase the toxicity of Hg. PMID- 2554541 TI - Novel synthesis of 2-fluoroestradiol from 19-nortestosterone: biomimetic oxidative defluorination to 2-hydroxyestradiol. AB - A new, short synthetic route to 2-fluoroestradiol from 19-nortestosterone is described which gives the target compound in an approximately 25% overall yield. Oxidative defluorination of 2-fluoroestradiol to 2-hydroxyestradiol via treatment with Fremy's salt/iodide ion is reported. This process is regarded as biomimetic with respect to cytochrome P-450-dependent oxidative defluorination. PMID- 2554542 TI - Effect of progestins, estradiol, and coenzymes NAD and NADPH on the interconversion of estradiol and estrone in rabbit uterus in vitro. AB - The biotransformation of estradiol (E2) and estrone (E1) in the uterus of rabbits treated with norgestrel (NG), norethindrone (NET), norethindrone acetate (NETA), progesterone (P4), and E2 either by subcutaneous injection in oil or by intrauterine steroid-releasing silastic implants was carried out under an in vitro short-term incubation system. The studies have shown that E2 stimulates 17 beta-hydroxysteroid dehydrogenase (17 beta-OHSD) much more than P4 as compared to untreated controls. The kinetic studies on E2 metabolism in the presence of added coenzyme NAD showed an initial rapid estrone formation and a gradual reconversion of E1 to E2. The addition of NADPH, ATP, and glucose-6-phosphate facilitates the reconversion of E1 to E2. The interconversion of E2 and estrone in the presence of coenzymes was five- to ten-fold higher in the endometrium than in the myometrium per milligram protein. Both E2 and progestins stimulate the uterine 17 beta-OHSD activity in rabbit uterus. This study further suggested that the hormone-induced metabolism of estradiol and estrone in the rabbit uterus is essentially modulated by the availability of coenzymes. PMID- 2554543 TI - Spectrum of neurologic events following cardiac transplantation. PMID- 2554544 TI - Small GTP-binding proteins. PMID- 2554545 TI - Serum 5'nucleotidase and alkaline phosphatase--highly predictive liver function tests for the diagnosis of graft-versus-host disease in bone marrow transplant recipients. AB - The diagnostic efficacy of five serum liver function tests (aspartate and alanine aminotransferase, alkaline phosphatase, 5' nucleotidase, and bilirubin) was investigated in 95 bone marrow transplant recipients in whom acute graft-vs-host disease was graded by the Seattle criteria. The patient population included a control group of 22 autologous transplant recipients (group I), 33 patients with no GVHD (group II), 21 patients with grades 1 and 2 GVHD (group III), 12 patients with grade 3 GVHD (group IV), and 7 patients with grade 4 GVHD (group V). Student t test analysis of the analytes among the five groups of patients showed that 5' nucleotidase and alkaline phosphatase were the best discriminants among all the possible combinations of group pairs. Peak levels of 5' nucleotidase within each group of patients correlated well with those of alkaline phosphatase in all the allogeneic transplant groups (II-V; r = 0.59), but the correlation of these with bilirubin was less frequent. Also, 5' nucleotidase and alkaline phosphatase showed significant discrimination (P less than 0.05) even between groups I and II, suggesting that they are more sensitive than the Seattle criteria in the diagnosis of GVHD. They also showed the best overall discriminatory ability by one-factor analysis of variance (ANOVA; P = 0.0001 as compared with 0.002, 0.009, and 0.04 for aspartate aminotransferase, alanine aminotransferase, and bilirubin, respectively). Receiver-operating curves of the five analytes again revealed that 5' nucleotidase and alkaline phosphatase were by far the best discriminators among the five groups of patients. Bilirubin was relatively insensitive because it was a good discriminator only between the control group and groups IV and V. The hepatocellular enzymes, alanine and aspartate aminotransferase, correlated well (r = 0.80) but discriminated poorly among the four groups of allogeneic transplant recipients (II-V), suggesting that all four groups had some measure of hepatocellular damage that was independent of the severity of GVHD. PMID- 2554546 TI - The role of CMV immune prophylaxis in patients at risk of primary CMV infection following orthotopic liver transplantation. PMID- 2554547 TI - Enterotoxin induced diarrhoea--an update. AB - The pathogenic personality or the criteria required to be a successful pathogen, of enteric bacteria includes, among others, the ability to produce potent proteins which by different intracellular mechanisms elicit what we overtly see as diarrhoea. Enteropathogens belonging to several genera like Vibrio, Escherichia, Shigella, Salmonella, Campylobacter, Aeromonas and Yersinia include species capable of elaborating strikingly similar exotoxins which seem to share common mechanisms of action involving specific receptor binding, internalization of the toxin followed by interaction with an intracellular target. It is now clear that there are several families of structurally, functionally and immunologically identical bacterial enterotoxins. In this communication, we have reviewed the recent developments on the various families of structurally homologous and antigenically cross reacting enteric toxins. PMID- 2554548 TI - Membranous obstruction of the inferior vena cava and invasion of the inferior vena cava by tumour in hepatocellular carcinoma. AB - A 28 yr old Zulu presented with a painful swelling in the right hypochondrium and severe swelling of the legs of short duration. The serum alpha-fetoprotein concentration was over 2 X 10(5) ng/ml and imaging showed a large hepatic mass lesion. Radionuclide venography revealed no flow through the inferior vena cava but flow through a large collateral vessel. Contrast venography showed the upper portion of the inferior vena cava to be occluded: large collateral vessels arose from the lower vena cava and the iliac veins. The histological features were those of longstanding hepatic venous outflow obstruction with irregular centrizonal and portal fibrosis: severe acute centrizonal congestion was not seen. This combination of findings indicates the presence of both membranous obstruction of the inferior vena cava, a rare developmental abnormality which predisposes to hepatocellular carcinoma formation, and invasion by the tumour of the inferior vena cava via the hepatic veins, an uncommon complication of hepatocellular carcinoma. PMID- 2554549 TI - In vitro immune-receptor gene rearrangements in clonal thymic lymphomas induced by Abelson murine leukemia virus. AB - The Abelson and Moloney murine leukemia virus complex (A-MuLV/M-MuLV) induces rapidly growing thymic lymphomas following direct injection into the thymus of newborn BALB/c and C57BL/6 mice. Southern blot analysis with a v-abl specific probe not only demonstrated that primary tumors are clonal, but also that the pattern of A-MuLV provirus integration is quite stable in primary tumor cells, as well as in their derived cell lines and clones. Most of the cell samples were able to rearrange the immunoglobulin heavy chain genes in culture, whereas in two cases the T cell receptor gamma chain genes also underwent rearrangement. Since the recombination mechanism is operative only in very immature lymphoid cells, these data provide indirect evidence for the lack of differentiation of A-MuLV cell targets in the thymus. PMID- 2554550 TI - Is the Epstein-Barr virus involved in Hodgkin's disease? AB - EBV genomes, in clonal episomal form, were detected in 7 out of 17 cases of Hodgkin's disease (HD) and in a single case of non-Hodgkin's lymphoma which occurred in a patient after therapeutic treatment for HD. The experimental data presented imply that a clonal cell population, harboring the EBV genome, must be present in EBV-positive HD. In light of this finding we are attempting to reconsider the abundant literature on this lymphoproliferative disorder, and suggest a reevaluation of the possibility that EBV could be etiologically involved in HD. PMID- 2554551 TI - Kinetics and regulation of tyrosine phosphorylation of the platelet derived growth factor receptor. AB - Platelet derived growth factor (PDGF) interaction with the cells induces rapid tyrosine phosphorylation of the PDGF receptor in a dose dependent manner. At 37 degrees C phosphorylation of the receptor is followed by its dephosphorylation and internalization. It is observed that the higher the ligand concentration, the more transient is the response, and the observed kinetics are explained by a simple kinetic model. At 4 degrees C the phosphorylated form of the receptor is more stable; however, if PDGF is dissociated from the cell surface-associated ligand-receptor complexes, the receptors are rapidly dephosphorylated, indicating that phosphatases specific for phosphotyrosine groups are very active within the cells. In fact, addition of orthovanadate stabilizes the phosphorylated form of the receptor and helps in recognizing possible physiological substrates of the PDGF receptor kinase. The expression of PDGF receptors on the cell surface has been investigated under different growth conditions: a positive correlation exists between the amount of PDGF receptors and the duplication times of exponentially growing cultures. Moreover, during exponential growth the PDGF receptors are scarcely expressed, and their number increases reaching a maximal value when the population enters the stationary phase. PMID- 2554552 TI - Relationship between CaMBr1 expression and tumor progression in small cell lung carcinomas. AB - In order to study the possible relationship between antigenic phenotype and tumor progression, 63 small cell lung carcinomas (SCLC) biopsies derived from primary or metastatic tumors were tested by immunofluorescence and immunoperoxidase techniques with an anti-carcinoma monoclonal antibody designated MBr1. Primary tumors were found to be less reactive with MBr1 than the local and distant metastatic lesions (57% versus 75% and 89% positivity respectively). A life table analysis on the tested cases indicated an inverse association between the expression of the marker recognized by the MBr1 MAb (CaMBr1) and overall survival (p less than 0.01): patients with MBr1-positive tumors showed a shorter survival time in comparison to patients whose tumors did not express the marker. The same correlation between survival and CaMBr1 expression was found even when only the 31 cases of early stage disease patients were considered (p less than 0.05). Different tumor aggressiveness or resistance to therapy of MBr1-positive tumors could be responsible for the shorter survival time of the patients. PMID- 2554553 TI - [Carcinoma in situ of the testis: clinical significance, diagnosis and therapy]. AB - The term "carcinoma in situ (CIS) of the testis" is used to mean the presence of atypical neoplastic spermatogonia exhibiting distinct morphological and immunohistological characteristics that differentiate them from normal germ cells. The lesion is considered to be the uniform precursor cell of all germ cell tumors, and it can usually be detected many years before manifestation of the tumor by surgical biopsy and subsequent immunohistological staining for placental alkaline phosphatase (PIAP). This paper gives a review of the theoretical and clinical features of CIS. The description is based on data in the literature and on the authors' experience with two recently detected cases of CIS in the contralateral testis of patients with germ cell tumors. In view of the observation of intraepithelial spread inside the seminiferous tubules, the term "testicular intraepithelial neoplasia" (TIN) is suggested instead of carcinoma in situ. PMID- 2554554 TI - [Effectiveness of HBO in the complex treatment of patients with infectious destructive lung diseases]. PMID- 2554555 TI - [Sorption characteristics of natural zeolite (clinoptilolite) in biological material in vitro]. AB - The zeolite (clinoptilolite) sorption of arsenic, cadmium, and lead ions from rumen fluid and abomasum juice was investigated in laboratory conditions. Zeolite was found to sorb 91% of lead and 45% of cadmium from rumen fluid in 24 hours. The sorption effectiveness was even higher from abomasum juice where zeolite sorbed 98% lead in 24 hours. PMID- 2554556 TI - Prevalence of feline leukaemia virus and antibodies to feline immunodeficiency virus in cats in the United Kingdom. AB - A representative sample of the pet cat population of the United Kingdom was surveyed. Blood samples from 1204 sick and 1007 healthy cats of known breed, age and sex were tested for antibodies to feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV). The prevalence of FIV was 19 per cent in sick cats and 6 per cent in healthy cats, and the prevalence of FeLV was 18 per cent in sick cats and 5 per cent in healthy cats; both infections were more common in domestic cats than in pedigree cats. Feline immunodeficiency virus was more prevalent in older cats but FeLV was more prevalent in younger cats. There was no difference between the prevalence of FeLV in male and female cats but male cats were more likely to be infected with FIV than female cats. No interaction was demonstrated between FIV and FeLV infections. Of the cats which were in contact with FIV in households with more than one cat, 21 per cent had seroconverted. The prevalence of FeLV viraemia in cats in contact with FeLV was 14 per cent. The clinical signs associated with FIV were pyrexia, gingivitis/stomatitis and respiratory signs, and with FeLV, pyrexia and anaemia. It was concluded that both viruses were significant causes of disease, and that the cats most likely to be infected with FIV were older, free-roaming male cats and for FeLV, younger, free roaming cats. PMID- 2554558 TI - Induction of virus-producing tumours in athymic nude mice by bovine papillomavirus type 4. AB - Bovine papillomavirus type 4 (BPV-4), the causative agent of alimentary papillomatosis, has been used to infect, in vitro, fragments of palatine mucosa from late term bovine fetuses. These small explants were placed beneath the renal capsule of athymic nude mice where they grew to produce, at first, squamous epithelial cysts containing BPV-4 genomic DNA and, later, papillomas which were morphologically identical to those of cattle and which contained large amounts of replicating virus. The possible utility of this technique in assessing neutralising antibodies in vaccine development is discussed. PMID- 2554557 TI - Clinical and laboratory findings in cats infected with feline immunodeficiency virus. AB - Thirty-two cats referred to the Feline Studies Centre between June 1987 and October 1988, and 14 in-contact cats, were found to be infected with feline immunodeficiency virus. Most of the 46 cats were non-pedigree and free ranging; 27 were male (19 neutered) and 19 were female (18 neutered). Their ages ranged from one to 17 years and the average age was 5.8 years. The most common clinical signs were lethargy, inappetence, weight loss, pyrexia and lymphadenopathy; most cases had multiple abnormalities. Other common signs were gingivitis, diarrhoea, rhinitis and ocular discharge. Eight cats had neoplasia. The commonest haematological abnormalities were anaemia, neutropenia, lymphopenia and monocytosis. Eight cats had lymphocytosis; seven of these were in a single house hold. Several cats had high serum globulin levels and half of those tested had high IgG levels. Seven cats had no detectable antibody to feline immunodeficiency virus even though the virus was cultured from the peripheral blood lymphocytes. During follow-up for up to 60 weeks one cat died and 23 were destroyed on humane grounds. PMID- 2554559 TI - [Macroaggregated calcifications in cancer of the breast]. AB - Breast cancer is characterized by the presence of multiple aggregated microcalcinates. However, the authors managed to detect 6 cases of breast cancer with macroaggregate calcifications, of which 3 were not adequately imaged on mammograms. Morphological investigations in all cases have shown that macroaggregate calcifications that are typical of benign lesions, are also detectable in breast cancer. The absence of an image of calcification on mammograms is probably accountable for by its various density. PMID- 2554560 TI - [Differential diagnosis of breast tumors with limited growth]. AB - The paper is concerned with the results of comparative analysis of the clinical, x-ray and cytological methods of investigation of women with breast tumors in situ (43 cases of cancer and 19 cases of intracanalicular fibroadenoma, the latter being difficult for differential diagnosis). A number of important diagnostic symptoms were identified. Cancer affected more frequently women over 55. It was characterized by the presence of microcalcinates (60 +/- 13%) and inadequacy of tumor sizes on palpation and on radiograms (35 +/- 7%). The diagnostic accuracy of the methods used separately was 44 +/- 8% for the clinical method, 79 +/- 7% for the x-ray method, and 89 +/- 3% for the cytological method. Their combined use in a specialized mammography unit resulted in 98 +/- 2% of diagnostic accuracy. PMID- 2554561 TI - [Informativeness of angiography in trophoblastic tumors of the uterus]. AB - A study was made of the resolving power of general and selective pelvic angiography with regard to specified diagnosis of trophoblastic tumors of the uterus. The informative value of general pelvic angiography was estimated by the results of angiomorphological correlations in 81 operated patients. The significance of selective pelvic angiography performed in 152 women was based on comparison of its results with those of pharmacoangiography supplemented by hysterography in 86 patients. The correctness of diagnosis in 26 of them was confirmed by operative findings. Analysis of the investigations has shown that general pelvic angiography allowed the detection of a tumor, its size and site in the uterus in 48% of the patients only. Selective pelvic angiography combined with pharmacoangiography was shown to be a method extending the diagnostic potentialities of pelvic angiography. In the authors' experience, it permitted obtaining a direct contrast image of a trophoblastic tumor or excluding its presence in the uterus of 85% of the examinees. PMID- 2554562 TI - Negative enrichment of bovine T lymphocytes with monoclonal antibodies and magnetic microspheres. AB - A highly enriched population of bovine T lymphocytes was produced from peripheral blood leukocytes following the depletion of monoclonal antibody-labelled B lymphocytes and monocytes with magnetic microspheres. This negative-enrichment protocol was simple, rapid, and specific. Also, it had a high recovery efficiency and was consistently reproducible. The enriched T lymphocytes proliferated in response to recombinant bovine interleukin 2 and, following the addition of monocytes, to concanavalin A. This methodology made it possible to determine the proliferative responses of peripheral blood lymphocytes utilizing a constant number of T lymphocytes within each assay. In this way, the in vitro T lymphocyte responses were determined independent of changes in the number of responder cells within peripheral blood. PMID- 2554563 TI - Insulin and C-peptide co-localization in the beta granules of normal human pancreas and insulinomas. A quantitative immunocytochemical approach. AB - It has been shown, by using the immunogold technique, that C-peptide and insulin are co-localized in the mature granules of human pancreatic beta cells and insulinomas with typical granules. The mean gold bead densities of both C-peptide and insulin were at least twice as high in the normal pancreas when compared with the insulinomas. The mean granule diameter of the insulinoma cells (D = 0.30 +/- 0.12 micron) was smaller than that of human pancreatic cells (D = 0.45 +/- 0.15 micron). The morphometric data indicate that each of the antigens (C-peptide and insulin) is distributed similarly in the halos and the dense cores of the beta granules. Thus, no topological segregation of these two antigens occurs within the beta granules of either normal human pancreas or insulinomas. PMID- 2554564 TI - Cytophotometric DNA analysis of hepatocellular carcinoma with Mallory bodies. AB - In order to clarify the pathological significance of Mallory body (MB) formation in human hepatocellular carcinoma (HCC), cell nuclear deoxyribonucleic acid (DNA) content was measured microspectrophotometrically in 20 autopsied cases of HCC associated with cirrhosis and bearing many MBs. According to the degree of dispersion, the DNA histogram was classified into type I (diploid pattern), type II (hyperploid pattern) and type III (aneuploid pattern). Non-neoplastic hepatocytes of normal livers and of cirrhotic areas of the 20 HCC cases showed generally a diploid pattern (type I). In contrast, MB-positive HCC cells showed more hyperploidy or aneuploidy (type I: 0%; type II: 35%; and type III: 65%) compared with MB-negative HCC cells (type I: 25%; type II: 50%; and type III: 25%). These data suggest that MB formation in HCC is accompanied by a constant change of DNA content of HCC cells, though the causal relation between them is only speculative. Two separate HCC nodules in the same liver, both of which contained many MB-positive cells, showed the same type of DNA histogram pattern, suggesting the possibility that they were of a monoclonal origin and had spread discontinuously in the liver. PMID- 2554565 TI - Mutagenesis of an AUG codon in the adeno-associated virus rep gene: effects on viral DNA replication. AB - The adeno-associated virus (AAV) rep gene is transcribed from two promoters, p5 and p19, which code for two over-lapping families of rep proteins. The proteins coded by p5 transcripts contain an amino-terminal domain not present in the proteins coded by p19 transcripts. The rep gene is required for AAV DNA replication and also mediates pleiotropic effects in positive and negative regulation of expression of genes driven by either AAV or heterologous promoters. All three functions require rep proteins coded by p5 transcripts. The functions of the rep proteins coded by the p19 transcripts could not be independently discerned since the coding region for these proteins and the p19 promoter are embedded within the coding region of the p5 transcript unit. We describe here an AAV mutant in which the putative AUG initiation codon in the p19 transcripts was altered and which did not express p19-coded rep proteins. This mutant exhibited normal AAV duplex RF DNA replication but was deficient in accumulation of AAV single-stranded progeny DNA and infectious AAV particles. This mutant defines a novel phenotype for a rep gene mutation and suggests a role for the rep proteins in the generation or accumulation of the viral SS DNA. Moreover this mutant distinguishes two different functions of the rep protein(s) in accumulation of the RF molecules and accumulation or processing of the SS DNA molecules, respectively. PMID- 2554566 TI - The SV40 nucleosome-free region is detected throughout the virus life cycle. AB - The structures of SV40 intracellular chromatin complexes and of extracellular virus particles were examined by photolabeling with a radioactive psoralen derivative in order to determine the fate of the exposed origin region during the virus life cycle. We have previously shown that the origin region of intracellular SV40 chromatin is preferentially accessible to psoralen derivatives in vivo, whereas psoralen adducts are uniformly distributed when purified virus particles are photoreacted. We demonstrate here that when virion is photoreacted prior to a freeze-thaw cycle, the exposed regulatory region detected in intracellular nucleoprotein complexes is also found in mature virus particles. In contrast, if the virion is frozen and thawed prior to the photoreaction, the origin is not preferentially exposed to photoaddition. Virus particles that have not been subjected to a freeze-thaw cycle were found to exhibit preferential labeling in the origin region whether they were irradiated intracellularly, in culture medium, or following purification. Banding the virus in CsCl had no significant effect on the relative accessibility of the origin region to psorealen. Our findings indicate that the open regulatory region found on intracellular SV40 chromatin persists throughout the virus life cycle. PMID- 2554567 TI - Biochemical characterization of cell-associated and extracellular products of the Friend spleen focus-forming virus env gene. AB - The mature product of the env gene of Friend spleen focus-forming viruses (F SFFV) is efficiently released from both leukemia cells and infected fibroblasts. Analyses of the kinetics of env protein synthesis and secretion in NRK cells infected with the Lilly-Steeves strain of SFFVp indicated that this product, gp65, was formed rapidly and remained stably associated with cells for up to 4 hr, at which point it was first detected in supernatant medium. By 12 hr after synthesis, greater than 95% of gp65 was found extracellularly. The release of this component was effectively blocked by 10 mM 1-deoxynojirimycin, an inhibitor of oligosaccharide processing, demonstrating a requirement for processing of high mannose precursor oligosaccharides in the secretion of gp65. Similar oligosaccharide substituents were found on cell-associated and extracellular forms of gp65. Enzymatic deglycosylation experiments demonstrated that in addition to the predicted four N-linked oligosaccharides, gp65 contains O-linked carbohydrates which are resistant to the action of peptide N-Glycanase F, but sensitive to neuraminidase and O-Glycanase. These structures may be related to O linked oligosaccharides previously found on the env gene products of murine leukemia viruses. Comparison of the sizes of the deglycosylated forms of cell associated and supernatant gp65 demonstrated that the extracellular molecules are approximately 3 kDa smaller than the cell-associated components. These data suggest the involvement of proteolysis at a C-terminal site in the release of gp65 from the plasma membrane. PMID- 2554568 TI - Vimentin phosphorylation by p37mos protein kinase in vitro and generation of a 50 kDa cleavage product in v-mos-transformed cells. AB - Previous studies have shown that vimentin, an intermediate filament protein, is reduced in amount in cells acutely infected with Moloney mouse sarcoma virus (Mo MuSV). In this report, we provide evidence for specific alteration of vimentin in Mo-MuSV-transformed cells and demonstrate specific phosphorylation of vimentin by the p37mos protein kinase in vitro. Specificity of the phosphorylation reaction was demonstrated by using viral mos proteins encoded by various isolates of Mo MuSV and p37mos produced in yeast. A phosphotransfer domain mutant lacking the ability to autophosphorylate p37mos failed to phosphorylate vimentin. Similarly, vimentin was not phosphorylated by the temperature-sensitive P85gag-mos kinase derived from infected cells maintained at the restrictive temperature. In ts110 MuSV-transformed NRK cells, vimentin was phosphorylated at both the permissive and nonpermissive temperatures for transformation. However, at the permissive temperature, an altered form of vimentin (about 50 kDa) with a more basic isoelectric point and lower apparent molecular weight was detected. This 50-kDa product was highly phosphorylated as compared to the bulk of the normal 55-kDa form of vimentin. On the basis of its mobility in two-dimensional gels, the 50 kDa form of vimentin should lack the carboxy terminus. This type of alteration could conceivably modulate the function of vimentin filaments in the transformed cell. PMID- 2554569 TI - Expression of avian reticuloendotheliosis virus envelope confers host resistance. AB - We constructed two reticuloendotheliosis virus (REV) envelope gene expression plasmids, one containing the REV-A envelope gene, the other the spleen necrosis virus (SNV) envelope gene. Cell lines were generated by transfecting each of the REV envelope plasmids into D17 cells, a canine cell line. The levels of REV envelope glycoprotein in the cell lines were assayed by immunoprecipitating the envelope glycoproteins from lysates of cells that were labeled with [35S]methionine. Virological challenge assays determined the degree of resistance of each of the cell lines to REV-A or SNV infection. The expression of either envelope gene protected the cells from infection by either REV-A or SNV virus. Several cell lines were significantly more resistant to REV infection than the parental D17 cells, and two lines were 25,000-fold more resistant, approaching the resistance of REV-infected D17 cells to reinfection. The resistant cell lines were not able to confer resistance to susceptible cells by cocultivation. The level of resistance was correlated with the uniformity of expression of the REV envelope glycoproteins by the individual cells in a cell line and not with the absolute level of expression by the population of cells. PMID- 2554570 TI - The release of bluetongue virus from infected cells and their superinfection by progeny virus. AB - Immunoelectron microscopy using an anti-VP2 monoclonal antibody complexed to colloidal gold has been used to study the mechanism of bluetongue virus (BTV) release from infected cells. Examination of the BTV-infected cell surface revealed that viruses are released both as enveloped particles, by budding through the plasma membrane, and as nonenveloped particles by "extrusion" through the membrane. Particles being released and those remaining on the cell surface retain an association with the cortical layer of the cytoskeleton. Analyses of virus particles released from infected cells and the intracellular viruses in the cytosol and attached to the cytoskeleton indicate that although the three populations have similar particle to infectivity ratios they differ in their ability to bind gold-labeled anti-VP2 antibody. The fact that released viruses bind less antibody than intracellular viruses suggests that virus release from infected cells may be associated with either a loss of VP2 or a rearrangement of the virus outer coat which obscures a proportion of the reactive epitopes on the virus surface. Electron microscopic observations also indicated that, in addition to virus release, events at the plasma membrane resulted in the uptake of progeny virus by endocytosis. Elevation of intraendosomal/lysosomal pH by lysomotropic bases and an acidic ionophore inhibited BTV replication when added to cells concurrently with the virus. Addition of such agents to infected cells at 4 hr p.i. decreased both the maximum titer of released virus and the rate at which virus antigen was synthesized in infected cells. Addition of anti-BTV antiserum 4 hr p.i. also resulted in a decreased rate of intracellular virus antigen accumulation. These results suggest that superinfection of BTV-infected cells by progeny virions effectively increases the multiplicity of infection and enhances the kinetics of BTV replication. PMID- 2554571 TI - Demonstration of the human herpesvirus 6-induced DNA polymerase and DNase. AB - Infection of HSB-2 cells with human herpesvirus 6 (HHV6) results in an approximately 51-fold increase in the level of DNA polymerase activity and a 4.44 fold increase in the level of DNase activity when compared to mock-infected cells. There was no increase in thymidine kinase, uracil-DNA glycosylase, or deoxyuridine triphosphate nucleotidohydrolase activities in the infected cells. The HHV6-induced DNase and DNA polymerase activities could be distinguished from their normal cellular counterparts on the basis of immunological specificities and in the case of DNA polymerase based upon differences in electrophoretic migration. Serological studies also demonstrated reactivity of the antisera not only for HHV6 but also for Epstein-Barr virus. PMID- 2554572 TI - Murine cytomegalovirus-induced suppression of antigen-specific cytotoxic T lymphocyte maturation. AB - Antigen-specific cytotoxic T lymphocyte (CTL) maturation was inhibited in mice acutely infected with murine cytomegalovirus (MCMV). When immunization with Simian virus 40 (SV40) either preceded or followed infection with MCMV by 1 day, the frequency of SV40-specific CTL precursors among lymph node cells (LNC) was significantly reduced compared to noninfected mice. Replication of the herpesvirus in LNC could not be detected; however, MCMV rendered noninfectious by heat treatment was not suppressive to CTL development. Lymph node cells from nonimmunized, MCMV-infected mice contained a cell(s) present in low frequency which suppressed in vitro maturation of SV40 CTL in immune lymph nodes from mice not infected with MCMV. These suppressor cells did not affect the antigen- and interleukin-2-dependent growth nor cytotoxic activity of a mature, SV40-specific CTL clone. These results indicate that MCMV interferes with immunoregulatory functions required for development of antigen-specific CTL precursors to mature effector CTL. The immunosuppression is mediated at least in part by the MCMV induced suppressor cell(s). PMID- 2554573 TI - A herpes simplex virus ribonucleotide reductase deletion mutant is defective for productive acute and reactivatable latent infections of mice and for replication in mouse cells. AB - Herpes simplex virus encodes a ribonucleotide reductase that is not essential for virus growth in dividing cells at 37 degrees. This enzyme has been proposed as a target for antiviral drugs; its utility in this regard could depend upon its importance in vivo. To test the requirement of viral ribonucleotide reductase in a mammalian host, we tested a mutant virus, lacking most of the gene encoding the ribonucleotide reductase large subunit, in a mouse eye model of pathogenesis and latency where the wild-type virus establishes reactivatable latent infections in trigeminal ganglia following corneal inoculation. The deletion mutant was severely impaired in its ability to replicate acutely in the eye and in the trigeminal ganglion and failed to establish reactivatable latent infections. In contrast, a recombinant virus in which the deleted sequences were restored was competent for both acute and latent infections. The defects of the deletion mutant in the mouse may be related to its severely impaired growth at 38 degrees in mouse cells relative to its growth in Vero cells. These results indicate that ribonucleotide reductase is critical for productive acute and reactivatable latent infections in mice and replication in mouse cells at 38 degrees and suggest that caution be exercised in extrapolating from studies conducted in mice to human infections when judging the utility of this enzyme as a target for antiviral chemotherapy. PMID- 2554574 TI - Duplication of enhancer sequences in human papillomavirus 6 from condylomas of the mamilla. AB - Human papillomavirus (HPV) 6 usually induces tumors of the genital, oral, or laryngeal mucosa. An HPV 6-related DNA of 8.2 kb was detected in an extrachromosomal state in atypically located condylomas of the mamilla and was molecularly cloned. The identity of the cloned HPV DNA and the viral DNA in the biopsy was confirmed by comparative Pstl cleavage analysis, which showed typical HPV 6 DNA fragment patterns except for 0.2-kb larger B fragments. Sequencing revealed an exact 236-bp duplication encompassing nucleotides 7681 to 7896 of HPV 6b. This tandem repeat is just upstream from the putative early promoter and contains a 20-bp insertion at position 7720, which constitutes an enhancer element described by R. F. Rando, W. D. Lancaster, P. Han, and C. Lopez (Virology 155, 545-556, 1986). A Hinfl-Pstl fragment containing the whole duplication was cloned into an enhancer-dependent CAT expression vector and led to three- to sevenfold increased CAT activity when compared with the monomeric sequence in C127 cells and BPV1 transformed C127 cells. This indicates that the duplication within the HPV 6 isolate from the mamilla may influence early gene expression and possibly tissue tropism. PMID- 2554575 TI - Genome sequence of tick-borne encephalitis virus (Western subtype) and comparative analysis of nonstructural proteins with other flaviviruses. AB - The genome sequence of tick-borne encephalitis (TBE) virus (Western subtype vaccine strain Neudoerfl) was determined. This extends the previously published sequence of the structural proteins to the nonstructural protein region and noncoding sequences at the 5'- and 3'-termini. The amino-termini of the individual proteins were assigned by comparison with other flavivirus sequences. Amino acid homology calculations between TBE virus and mosquito-borne flaviviruses were performed for all nonstructural proteins. An evolutionary tree based on protein NS1 is presented that reveals the molecular basis of relationships among flaviviruses. Tick-borne and mosquito-borne flaviviruses share a common hydrophilicity profile and also other features of their primary sequences, such as the presumably functional Gly-Asp-Asp sequence element within protein NS5. Other characteristics, such as the potential N-glycosylation sites of protein NS1 and a potential proteolytic cleavage site within protein NS4B, are conserved within the mosquito-borne group, but differ in the TBE virus sequence. PMID- 2554576 TI - Synthesis of functional bovine leukemia virus (BLV) p34tax protein by recombinant baculoviruses. AB - The bovine leukemia virus (BLV) p34tax (also called tat, p34, XLOR gene product) is a 34-kDa polypeptide encoded in the 3'-terminal region of the virus. This protein is responsible for positive transcriptional trans-activation of promoter elements located within the BLV long-terminal repeat. We introduced the protein coding region of BLV p34tax into the genome of the baculovirus Autographa californica nuclear polyhedrosis virus. After infection of the insect Spodoptera frugiperda (SF9) cell line, this recombinant strain of baculovirus produced approximately 100 to 150 mg of p34tax per 2 X 10(9) cells. This protein, when introduced into mammalian fibroblasts by using a cell-to-cell fusion technique, functionally trans-activated the BLV long-terminal repeat. Analysis of 32P labeled proteins of SF9 cells expressing BLV tax by two-dimensional gel electrophoresis indicated that the BLV p34tax was phosphorylated. PMID- 2554577 TI - Specificity of enzyme-substrate interactions in foot-and-mouth disease virus polyprotein processing. AB - A series of transcripts derived from FMDV cDNA plasmids containing defined regions of the genome were translated in a rabbit reticulocyte lysate system. The products were analysed directly or following incubation with an FMDV-infected cell processing extract. Processing by the L proteinase at the L/1A cleavage site occurred when most of the P1-2A protein was absent. Substitution of sequences upstream of the 2C/3A cleavage site showed that the 3C proteinase was also able to cleave at an entirely novel cleavage site, apparently at K-I amino acid pairs. Cleavage at the 2A/2B site was not only independent of L and 3C proteinases, but was shown to occur when 2A and as few as four 2B N-terminal amino acids were present. Thus, the disparate proteolytic activities responsible for all three primary processing events that give rise to the products L, P1-2A, 2BC, and P3 were highly resistant either to major deletion or substitution of protein sequences adjacent to, or at, the site of cleavage. By contrast, secondary processing in trans was sensitive to changes at remote sites. For example, removal of the C-terminal regions of P1-2A and 2BC precursors impaired their ability to act as substrates for 3C proteinase activity. Processing of P1-2A, particularly of the 1D/2A cleavage site, was enhanced by inclusion of sequences from the 3D region of the genome. PMID- 2554578 TI - Nucleotide sequence of bovine herpesvirus type 1 glycoprotein gIII, a structural model for gIII as a new member of the immunoglobulin superfamily, and implications for the homologous glycoproteins of other herpesviruses. AB - The gene encoding bovine herpesvirus type 1 (BHV-1) glycoprotein gIII was mapped, cloned, and sequenced. The gene is situated between map units 0.122 and 0.135 and encodes a predicted protein of 521 amino acids. The identity of the sequenced gene has been verified previously by expression of immunologically authentic recombinant BHV-1 gIII (Fitzpatrick et al. (1988) J. Virol. 62, 4239-4248). Comparison of the BHV-1 gIII amino acid sequence with the homologous glycoproteins of other alphaherpesviruses revealed significant homology in the carboxy-terminal half of the molecules, including six invariant cysteine residues. A 96 amino acid domain with significant homology to class II major histocompatibility complex (MHC) antigen constant domains was identified in the conserved carboxy-terminal half of BHV-1 gIII. This domain is flanked by two other similarly sized domains which may be related to other immunoglobulin (Ig) superfamily domains. These homologies support a model for the structure of BHV-1 gIII as a new member of the Ig superfamily. Elements of the model may be applicable to the homologous glycoproteins of other herpesviruses and relevant to the immunobiology of herpesvirus infections. PMID- 2554579 TI - Nucleotide sequence and mode of transmission of the wild mouse ecotropic virus, HoMuLV. AB - HoMuLV is an NB-tropic wild mouse leukemia virus (MuLV) with ecotropic host range which induces lymphoma, erythroleukemia, and myelogenous leukemia in NIH Swiss mice. Although HoMuLV uses the same cell surface receptor as other ecotropic MuLVs, hybridization studies suggested that the HoMuLV envelope glycoprotein differs from that of other ecotropic MuLVs. We have now molecularly cloned HoMuLV and sequenced its LTR, gag, and env regions. HoMuLV differs markedly from other MuLVs in the LTR U3 region, in the SU protein of env, and in p12gag. U3 contains a single copy of a sequence analogous to the direct repeat found in other LTRs, and this region includes several previously defined protein binding sites. The predicted amino acid sequence for the coding regions of env and gag reveal that p12 and the SU protein show less than 59 and 65% sequence identity, respectively, with those of other MuLVs. A 0.6-kb segment of the 5' region of the HoMuLV env was used as a hybridization probe to examine inbred and wild mouse genomic DNAs for proviral sequences. HoMuLV env sequences were not present in the germline of any of the inbred strains or wild mice examined including the Mus hortulanus mice which harbored infectious virus. Therefore, HoMuLV represents an evolutionarily related, but distinct, subgroup of ecotropic MuLV which is not genetically transmitted in its natural host. PMID- 2554580 TI - Persistence of varicella-zoster virus DNA in blood mononuclear cells of patients with varicella or zoster. AB - Varicella-zoster virus (VZV) DNA was detectable by in-situ hybridization in blood mononuclear cells (MNCs) of patients with varicella or zoster for 2-56 days after the onset of a rash. VZV DNA was present in many MNCs from one acute varicella patient 2 days after the onset of the rash and was rarely found in MNCs during acute zoster, convalescent zoster, and convalescent varicella. The morphology of MNCs containing VZV was heterogenous, although most viral-DNA-containing MNCs were large monocytoid cells. Serial examination of blood MNCs from one adult with varicella revealed VZV DNA up until 8 weeks, but not 16 weeks, after the appearance of the rash; parallel studies in four zoster patients showed VZV DNA up until 3 weeks, but not later than 7 weeks after the appearance of the rash. These results indicate that MNCs become infected with VZV during the primary encounter with VZV (varicella) and during reactivation (zoster) and that infection continues for weeks after the onset of the skin rash. Furthermore, the detection of VZV DNA in blood MNCs of uncomplicated zoster patients coincides with the period during which these patients experience pain. PMID- 2554581 TI - Molecular cloning of DNA sequences from cervical intraepithelial neoplasia that hybridize to human cytomegalovirus DNA. AB - We have prepared EMBL3 libraries of DNA extracted from the cervix of a patient with cervical intraepithelial neoplasia (CIN) and isolated seven recombinant clones containing sequences that hybridize to human cytomegalovirus (HCMV) DNA. Restriction analysis of one clone with BamHI and SalI endonucleases revealed that the insert DNA showed a high degree of homology to the HCMV Ad169 genome over the region between the HindIII K/E site and the SalI site located within the BamHI P fragment. The HCMV insert in the CIN clone is integrated and flanked by cellular sequences. The major immediate early gene that encodes a polypeptide of approximately 69 kD was found to be conserved in the CIN clone. Transfection of clones encoding the immediate early region of HCMV resulted in cells that were positive in immunofluorescence studies with two monoclonal antibodies directed against the HCMV 69 kD immediate early polypeptide. Infection of human ectocervical cells with HCMV Ad169 revealed that they could express the 69 kD polypeptide encoded by the immediate early gene but could not replicate the virus, whereas HCMV was able to replicate productively in cultured endocervical cells. HCMV has been shown to activate endogenous retroviruses and also to transcriptionally activate the long terminal repeat of human immunodeficiency virus. Activation of virus and cellular genes by HCMV may be a means by which this virus is involved in the multistage process of oncogenesis and/or the activation of latent infections. PMID- 2554582 TI - Nucleotide sequence of the pseudorabies virus immediate early gene, encoding a strong transactivator protein. AB - We report the complete DNA sequence of teh pseudorabies virus (PRV) immediate early (IE) gene and its flanking nucleotide sequences, together comprising 5091 base pairs. An open reading frame starts with an ATG codon in position 263 from the transcription-initiation site and ends with a TGA codon in position 4601, thus encoding a predicted protein of 1446 amino acids (150 kD). The PRV IE protein exhibits significant homology with the functionally related transactivator proteins, ICP4 of herpes simplex virus-1 (HSV-1) and p140 of varicella zoster virus (VZV). The extent of homology varies widely along the three sequences: Two regions of the PRV IE protein extending from amino acids 482 to 659 and 959 to 1350 exhibit 50% to 60% identity with the cognate sequences, whereas the remaining sequence reveals little homology apart from a common polyserine stretch. The base composition of the PRV IE coding region is 80% G + C, compared with 81.5% for HSV-1 and 64.1% for VZV. Yet the PRV IE protein appears to be as closely related to VZV p140 as to HSV-1 ICP4. The regions of strong homology are also apparent in plots predicting secondary structure. PMID- 2554583 TI - A temperature-sensitive defect of enterovirus 70 is located at the uridylylation of the genome-linked protein VPg in vitro. AB - The temperature-sensitive phenotype of enterovirus 70 (EV70) was examined by use of an in-vitro RNA replication system derived from a membrane fraction (crude replication complex, CRC) of EV70-infected HeLa cells. This system was capable of synthesizing the nucleotidyl proteins VPg-pU and VPg-pUpU. Formation of these nucleotidyl proteins was completely abolished when the in-vitro reaction was performed at the nonpermissive temperature for virus replication. Considering our previous observation that the defective stage of the temperature-sensitive growth of EV70 resides in the initiation step of RNA transcription in vivo, it is most likely that the lack of uridylylation of VPg at the restricted temperature in vitro is directly involved in the temperature-sensitive defect of virus growth in vivo. PMID- 2554584 TI - Human topoisomerase 1 messenger RNA is not destabilized by the herpes simplex virus type 2 virion-associated shut-off function. AB - A cDNA for human topoisomerase I (Topo 1) was used to identify a 4.1 kb polyadenylated Topo 1 mRNA in methotrexate-resistant human KB cells that are permissive for herpes simplex virus type 2 (HSV-2) infection. Using these cells, no effect of the HSV-2-associated early shut-off function on levels of Topo-1 mRNA was observed up to 6 hours postinfection, whereas the actin mRNA level was 22% cellular transcripts are susceptible. The level of several host-cell polyadenylated RNAs detected as cDNA clones (class 3 transcripts) were unchanged 8 hours after HSV-2 infection, and other cellular transcripts (class 2) actually accumulated at postinfection. PMID- 2554586 TI - Serological probes for some foot-and-mouth disease virus nonstructural proteins. AB - Foot-and-mouth disease virus (FMDV) O1 Kaufbeuren-specific cDNA fragments were subcloned into the E. coli expression vector pRIT.2T. Fusion proteins thus produced in bacteria were purified by affinity chromatography and inoculated into rabbits. Three sera thus obtained were found to be monospecific for FMDV proteins 3A, 3C, and 3D, respectively. Two others were prevalently directed against protein 2C, but in addition, either to protein 2B or to protein 3A. Five out of six mature nonstructural virus proteins can therefore be separately investigated in FMDV-infected cells, either by indirect immunofluorescence or by radioimmunoprecipitation. Immunofluorescence shows all investigated proteins to be located exclusively in the cytoplasm. One of them, protein 2C, transiently forms aggregates at the periphery of cells. Radioimmunoprecipitation confirmed current knowledge on maturation of FMDV proteins. It was further used to characterize postinfectional sera with regard to FMDV-specific antibodies. Cattle and guinea pig were found to have responded differently to FMDV nonstructural antigens. Furthermore, antigenicity of yet to be described FMDV polypeptides was observed in the guinea pig. PMID- 2554585 TI - Structural analysis of human papillomavirus type 6c isolates from condyloma acuminatum and juvenile-onset and adult-onset laryngeal papillomata. AB - The human papillomavirus type 6c (HPV-6c) genome was molecularly cloned from biopsy specimens of a juvenile-onset and an adult-onset respiratory-tract papillomata and a condyloma acuminatum of the cervix. To determine if the genital tract isolate and respiratory-tract isolates contain divergent sequences that may account for a difference in tissue trophism or for a difference in the age of onset of the disease, fine-structure mapping, heteroduplex analysis by electron microscopy, and nucleotide sequencing were used to examine the sequence relationship among these HPV-6c isolates. No differences were found in the digestion patterns with 23 restriction enzymes. Heteroduplex analysis among the three genomes demonstrated that they were colinear without apparent deletions or rearrangements and had greater than 90% sequence identity. In heteroduplex analyses with a different subtype (HPV-6e) that was molecularly cloned from a genital wart, the genomes were colinear with greater than 90% sequence identity over 90% of their length. The most divergent region had 75-80% sequence identity and was localized to the part of the genome containing the E5a and E5b open reading frames (ORFs). Comparison of the sequence of 1430 nucleotides in this region for two of the HPV-6c isolates did not identify any differences between them. Comparison with the published sequences of HPV-6b identified deletions/insertions and base changes with approximately 75% sequence identity, and comparison with HPV-11 identified only six base changes. Conservation of sequences in the E4-E5 region and similarity in the restriction enzyme maps demonstrated that HPV-6c and HPV-11 are independent isolates of the same HPV-6 subtype. PMID- 2554587 TI - Tumorigenic conversion of NIH 3T3 cells by transfection with Shope fibroma virus DNA. AB - The properties of NIH 3T3 cells transfected with Shope fibroma virus (SFV) DNA were investigated. Six focus-derived cell lines were established that display the following properties: a) They all contained SFV DNA sequences at early passages, b) five of them also expressed SFV RNA at early passages and induced tumors in nude mice, and c) four lines were anchorage independent. Transfection with cloned DNA fragments containing terminal sequences did not induce foci, except with fragment C, which contains the growth-factor gene. However, these transfected cells failed to cause tumors, suggesting that the growth factor alone may not be responsible for tumorigenesis. PMID- 2554588 TI - The influence of the herpes simplex virus-1 DNA template environment on the regulation of gene expression. AB - To determine the role of the HSV-1 genome structure and environment on the regulation of gene expression, we constructed recombinant viruses containing a heterologous gene inserted into either the immediate early ICPO or late glycoprotein C (gC) genes of HSV-1. The heterologous gene consisted of the SV40 early promoter (without enhancer sequences) linked to the coding sequences for the bacterial chloramphenicol acetyl transferase (CAT). The expression of CAT was examined in Vero cells infected with either virus (named ICP0-CAT and Sph 6). For both recombinants, expression of CAT was not dependent upon prior viral protein synthesis. The kinetics of expression of CAT-specific mRNA resembled that of the HSV-1 genes into which CAT was inserted. Primer extension analysis revealed that the SV40 promoter is recognized and used when placed in cis in two different HSV 1 genome locations, and Northern hybridization experiments confirmed that the heterologous gene was expressed in the absence of prior viral protein synthesis. Therefore, this gene was not regulated as strictly as an HSV-1 gene, but was influenced by the environment into which it was placed, presumably by factors that are present when the normal viral gene is on. PMID- 2554589 TI - [Analysis of herpes simplex virus DNA using restriction endonucleases]. AB - DNA of herpes simplex virus was analysed by restriction endonucleases EcoRI, BgII and HindIII. By this method it is possible to differentiate type 1 and type 2 of viruses. DNA of herpes simplex virus was prepared according to the method of Kinnunen and his collaborators which is described in detail. PMID- 2554590 TI - [Lipid peroxidation processes and the dynamics of shifts in the cholesterol level in plasma and erythrocytes under the influence of low-frequency acoustic oscillations]. AB - Distinct alterations in lipid peroxidation, in content of alpha-tocopherol and cholesterol fractions were observed in rat blood plasma and erythrocyte membranes after long-term low-frequency acoustic oscillations (105 decibels, diurnally, within 3 months). Under these conditions activities of superoxide dismutase and erythrocyte Na+, K+-ATPase were also altered. The rate and direction of the enzymatic activity alterations correlated with the duration of the acoustic oscillation exposure and the patterns studied. Enrichment of blood plasma and erythrocyte membranes with alpha-tocopherol within 2-4 weeks of the acoustic oscillation is considered as an important phenomenon. PMID- 2554591 TI - [Lipid peroxidation and ATPase activity in synaptosomal and mitochondrial fractions of the brain in hypoxia]. AB - Acute hypoxia was accompanied by intensification of lipid peroxidation in synaptosomal and mitochondrial rat brain fractions as well as by inhibition of Na+, K+- and Mg2+-ATPases. Preadministration of antioxidants vitamin E and ionol into animals prevented distinctly the increase in lipid peroxidation rate, while the ATPases activity was unaltered. Inhibition of the ATPases may be considered as a result of impairing effect of the lipid peroxidation products, which appears to be of importance for structure-functional deteriorations of the nervous system in hypoxia. PMID- 2554592 TI - [Cyclic AMP and cyclic GMP level in cells of spontaneously aborted fetuses with trisomic and triploid karyotypes]. AB - Contents of intracellular cAMP and cGMP were studied in cultivated fibroblasts from 4 spontaneous abortuses with trisomy 7 and trisomy 9 as well as in 4 triploid cell strains. Six diploid cell strains from medical abortuses were used as controls. Distinct increase in cAMP content both calculated per I mg of cell protein or per a cell was found in trisomic strains, whereas the cAMP content increase in triploid strains was detected only per a cell. The data obtained suggest that functions of membrane complexes were impaired in the cells with anomalous karyotype. PMID- 2554593 TI - [The role of cyclic nucleotides in reparative bone formation]. AB - Distinct alterations in content of cyclic nucleotides were observed during treatment of 133 patients with fractures of long tubular bones. The following effects were found after initial increase in cAMP content: increase in tissue permeability, aseptic inflammation, inhibition of cell mitosis, acceleration of cell enzymes synthesis, of protein biosynthesis and of the rate of microcirculation in the traumatized region. Elevation of cGMP concentration led to initiation of bone marrow proliferation, to augmented formation of organic matrix and its mineralization. Monitoring of cyclic nucleotides concentration enabled to evaluate the reparative osteogenesis activity. PMID- 2554594 TI - [Endoscopic evaluation of the therapeutic results in patients with small cell lung cancer]. AB - The analysis of the data of 352 fiber bronchoscopies performed in 142 cases of small-cell lung cancer before, in the course and after chemotherapy or chemoradiation treatment makes a case for complex application of X-ray and endoscopic procedures since the data on diagnosis and response to treatment obtained under the separate use of either procedure agreed in 67.7% and 80.3% of cases only. PMID- 2554596 TI - [Use of restriction enzyme analysis in parasitology]. AB - Application of analysis of nucleic acid using restriction enzymes in parasitology is discussed. The main advantage of the new technique is the possibility of direct and detailed studies at the level of DNA. At present, the "genetic probe" becomes more and more commonly applied to identification of both parasites and their transmitters. It appears that the technique of restriction analysis is of great significance for solving taxonomic problems in parasitology. PMID- 2554595 TI - [Assessment of an analysis of the connections of nutrition with risk factors for ischemic heart disease in a one-time epidemiologic study]. AB - Nutrition of 908 males aged 20-54 years, living in Tallinn, was studied in simultaneous population investigation by the method of one-day questioning. It was found that subjects with excessive body mass (EBM) and coronary heart disease (CHD), who did not keep to a diet, reduced their ration calorific value, consumption of saturated fatty acids, sugar, and increased consumption of polyunsaturated fatty acids and food fibres. No relations were detected between nutrition and CHD risk factors in the majority of subjects during the simultaneous population investigation. It has been suggested that subjects with EBM and CHD changed their nutrition habits due to the distribution of information among the population on the important role of nutrition in the development of EBM and atherosclerosis. PMID- 2554597 TI - Surgical approach to early gastric cancer with lymph node metastasis. AB - The clinicopathology of our series of patients (n = 486) with early gastric cancer was reviewed with special reference to metastasis to the regional lymph nodes. The incidence of lymph node metastasis was 15.8% (19/120) in patients with the protruded type of carcinoma and 11.7% (42/360) in patients with the depressed type of carcinoma. It was especially high in carcinomas of the IIa + IIc type of the former group. The incidence of lymph node involvement was higher in the group with larger tumors (greater than 30 mm) than in the group with smaller tumors (less than or equal to 30 mm). Metastatic lesions were detected in as many as 25.0% (16/64) of the cases with large tumors (greater than or equal to 50 mm); however, of the 46 cases with small tumors (less than 10 mm), 1 case had metastases to the nodes of group 1. There were 4 cases of m-cancer with metastasis to group 1 lymph nodes. Skip metastasis to group 2 lymph nodes was discovered in 4 cases. From these results, we feel that the standard operation for early gastric cancer is R2-gastrectomy including the complete removal of group 1 and 2 lymph nodes. R1-gastrectomy or local resection is thought to be sufficient for m-cancer with a lesion smaller than 10 mm in maximum diameter. PMID- 2554598 TI - Restrictive versus liberal blood transfusion policy for hepatectomies in cirrhotic patients. AB - To evaluate the worth of intra- and postoperative blood transfusion in cirrhotic patients undergoing resection for hepatocellular carcinoma, we compared 13 patients receiving transfusions and 14 matched contemporary patients who did not receive blood. Preoperative hematological and biochemical parameters, the type and extent of liver resection, and the mean blood loss (862 and 870 ml) were similar in the 2 groups. The total volume of intra- and postoperative blood transfused ranged from 400 to 1,800 ml (mean, 1,223 ml) in the patients receiving transfusions. During various postoperative time intervals, the mean values of hematocrit, hemoglobin, serum total bilirubin, and lactic dehydrogenase activity were significantly higher in the patients who were transfused compared to those who were not. Mean serum transaminase activities were similar in the 2 groups at the same times. The mean hematocrit values decreased from 36.8% preoperatively to a postoperative minimum of 27.0% in the transfused group, and from 39.9% to 26.1% in the nontransfused group. Our experience and theoretical reasons have led us to withhold blood transfusion until the hematocrit values fall below 30% during hepatectomy and below 20% in the postoperative period (or unless circulatory instability cannot be corrected otherwise). Fresh frozen plasma is preferred for volume substitution and, if blood has to be given, only up to 60-70% of estimated losses should be replaced by fresh blood. PMID- 2554600 TI - [Micromethod for the detection of antibodies for the serologic diagnosis of infectious mononucleosis]. PMID- 2554599 TI - Association of Epstein-Barr virus with an angioimmunoblastic lymphadenopathy-like lymphoproliferative syndrome. AB - A patient presented with fever, night sweats, generalized lymphadenopathy, cold agglutinin hemolytic anemia, and thrombocytopenia. Lymph node biopsy revealed a lymphoplasmacytic infiltrate sparing the cortical sinuses but associated with arborizing blood vessels. Epstein-Barr virus (EBV) DNA was demonstrated in tissue from an involved lymph node. These data suggest an association of EBV with a lymphoproliferative process that resembles angioimmunoblastic lymphadenopathy with dysproteinemia. PMID- 2554601 TI - Enzymatic histochemistry of the human umbilical vessels. AB - The enzymatic activities of human umbilical vessels from normal term pregnancies were studied by histochemical methods. The results obtained suggest that oxidative phosphorylation is of little importance, whereas pentose cycle and anaerobic glycolysis are probably the most important sources of energy in the umbilical vessels. The presence of leucine aminopeptidase activity in the venous endothelium probably represents a defensive mechanism inactivating oxytocin. PMID- 2554602 TI - Effect of N alpha-acetylmuramyl-L-alanyl-D-isoglutaminyl-N epsilon-stearoyl- L lysine on resistance to herpes simplex virus type-1 infection in cyclophosphamide treated mice. AB - The restoration of resistance by N alpha-acetylmuramyl-L-alanyl-D-isoglutaminyl-N epsilon-stearoyl-L-lysine [MDP-Lys(L18)] on herpes simplex virus (HSV) type-1 infection was examined in cyclophosphamide (CY)-treated mice. MDP-Lys(L18) was shown to restore the resistance to HSV infection in CY-treated mice when it was injected either subcutaneously, intravenously, or intraperitoneally before infection. Treatment with MDP-Lys(L18) in CY-treated mice restored impaired activity for inhibiting HSV growth in the liver. PMID- 2554604 TI - Nice work for everybody except the patient. PMID- 2554603 TI - Studies on the development of a medium with peptone and casein hydrolysate for the production of foot-and-mouth disease vaccine in BHK-21 cells. AB - Studies were undertaken to develop a cheaper medium with indigenous sources of peptone and casein hydrolysate for continuous culture of BHK-21 (suspension) cells and production of foot-and-mouth disease (FMD) vaccine. Eleven batches of experimental media were prepared using different indigenous sources of casein hydrolysate and peptone. These batches of media were tested for the growth of Razi BHK-21 cells in suspension and compared with the growth in control Eagle's medium. Out of eleven batches only four batches of the media produced cell growth equivalent to that with the Eagle's medium. Cells passaged in these batches of the experimental media supported virus growth and titres were comparable with those in Eagle's medium. Experimental batches of vaccine against FMD virus type A5 were prepared on a pilot scale as well as on a fermenter scale. The protection indices of these vaccine in guinea-pigs were satisfactory and induced 100% protection in cattle. One source each of peptone and casein hydrolysate was found suitable for the experimental medium. The medium developed is able to replace 10 out of 13 amino acids and also tryptose phosphate broth in Eagle's medium and is expected to reduce the cost of medium by 33% compared with Eagle's medium. PMID- 2554605 TI - Antigen presentation by B cells and macrophages of cytochrome c and its antigenic fragment when conjugated to the surface of liposomes. AB - An in vitro antigen presentation system was used to study how antigens coupled to the surface of phospholipid vesicles (liposomes) are presented to antigen specific T cells. Liposome-bound pigeon cytochrome c (PCC) was 30-40-fold more potent than free PCC when peritoneal macrophages were the presenting cell. B cells presented surface-bound PCC, albeit less efficiently than unmodified PCC. Surface-bound peptide epitope was presented by both cell types, but not as efficiently as unmodified peptide. With the T cell epitope, antigen processing was not required since glutaraldehyde fixed cells could present surface-bound peptide. PMID- 2554606 TI - Systemic and mucosal cell mediated immune responses against herpes simplex virus in vaccinated guinea-pigs detected by lymphocyte proliferation and monoclonal antibodies against macrophages and T cells. AB - Peripheral blood lymphocytes from guinea-pigs immunized with a herpes simplex virus (HSV) sub-unit vaccine (Skinner vaccine) were shown to produce an HSV antigen-specific lymphocyte proliferative response post vaccination. This response was associated with a reduction, compared with mock vaccinated and unvaccinated animals, in the clinical disease scores post intravaginal challenge. This reduction was not associated with the presence of macrophage or T cell infiltration in the vaginal mucosa postchallenge. In contrast mock vaccinated and unvaccinated guinea-pigs did not produce an HSV antigen-specific response post vaccination but did demonstrate HSV antigen-specific lymphocyte proliferative responses post intra-vaginal challenge. The mock and unvaccinated animals had higher clinical disease scores postchallenge compared with sub-unit vaccinated animals and this was associated with the presence of cellular infiltrates in the vaginal mucosa. These results are discussed in relation to the immunopathology of the disease. PMID- 2554607 TI - Protection and serum antibody responses in guinea-pigs and mice immunized with HSV-1 antigen preparations obtained using different detergents. AB - Guinea-pigs immunized with a zwitterionic detergent-extracted HSV-1 antigen preparation responded with EIA and NT serum antibody titres that were significantly greater than those elicited by a non-ionic detergent-extracted antigen preparation inoculated using a similar dosage schedule. Following intravaginal challenge of the guinea-pigs with HSV-2 (strain SH/B), there was no statistically significant difference in the protection afforded to these animals by the two antigen preparations, although the results indicated the zwitterionic detergent-extracted HSV-1 antigen preparation to be slightly superior in this respect. In mice, the zwitterionic detergent-extracted HSV-1 antigen preparation elicited an EIA antibody response and partial protection against homologous virus challenge. The relevance of these animal models to determination of immunogenicity and efficacy of HSV vaccines prepared for use in man, and the reasons for the differences in immunogenicity of these HSV-1 antigen preparations in guinea-pigs, are discussed. PMID- 2554608 TI - Immune response to immunostimulatory complexes (ISCOMs) prepared from human immunodeficiency virus type 1 (HIV-1) or the HIV-1 external envelope glycoprotein (gp120). AB - In mice, immunostimulatory complexes (ISCOMs) prepared from HIV-1 B external envelope glycoprotein (gp120) induced 10-fold higher antibody titres than gp120 emulsified in depot adjuvant, as measured by enzyme-linked immunosorbent assay (ELISA). Rhesus monkeys immunized with gp120 ISCOMs produced precipitating and virus neutralizing antibody titres equivalent to those seen in HIV-infected chimpanzees and humans. After multiple immunizations with HIV-1 B gp120 ISCOMs, a rhesus monkey developed a neutralizing response to the HIV-1 isolates RF and MN, but not to the CC isolate. Antisera from ISCOM-immunized rhesus monkeys recognized gp120 on the membranes of HIV-1 B-infected H9 cells, indicating the preservation of epitope structure in the ISCOMs matrix. PMID- 2554609 TI - The effect of ammonium chloride on the multiplication of herpes simplex virus type 1 in Vero cells. AB - The multiplication of herpes simplex virus type 1 (HSV-1) in Vero cells is inhibited by ammonium chloride. The formation of infectious virus was inhibited immediately after the addition of the agent into the culture fluid and was restored by removal of the agent. Although neither viral DNA replication nor nucleocapsid formation were affected by the addition of ammonium chloride at 4 h postinfection, the agent markedly inhibited the formation of enveloped particles and completely the formation of infectious progeny virus. These results indicate that one of the effects of ammonium chloride on the multiplication of HSV-1 is the inhibition of envelopment of viral nucleocapsids. In addition, the envelopment of HSV-1 nucleocapsids was inhibited immediately after the addition of monensin into the culture fluid. These findings suggest the importance of acidic pH of an intracellular compartment in the envelopment of HSV-1. PMID- 2554610 TI - Interferon-mediated inhibition of retroviral infection: use of a defective retrovirus carrying a drug-resistance gene. AB - We here report the results of an investigation of the effect of interferon on the establishment of new infections by a retrovirus. For this study, we used an infectious but replication-incompetent retrovirus carrying a drug-resistance gene and assayed for infectivity by measuring drug-resistant colony formation. Mouse interferon-beta inhibited retroviral infection of mouse CG1 cells in a dose dependent manner. However, a higher dose of interferon was needed for eliciting the antiretroviral effects than for action against vesicular stomatitis virus. The degree of antiretroviral effect was comparable over at least a 100-fold range of multiplicity of infection and the effect was most pronounced when the cells were continuously treated with interferon before infections and during infection and drug-selection. PMID- 2554611 TI - Acyclovir resistance in herpes simplex virus type 1: biochemical and functional studies on the thymidine kinase of the highly resistant R100 strain. AB - The biochemical and functional properties of the thymidine kinase (TK) of the herpes simplex virus type 1 mutant R100, that is highly resistant to 9-(2 hydroxyethoxymethyl)guanine (acyclovir), are reported in comparison with the properties of its parental strain, wt. The mutant induced the production of a TK activity that accounted for only 10% of the wt one. This feature was not apparently related to a defective expression of the TK gene but it was rather connected to some functional characteristics of R100 enzyme. Although affinities of this enzyme for ATP and thymidine were unchanged, apparent Vmax values for thymidine were much reduced. In addition, affinities for antiviral analogues acyclovir, 9-(1,3-dihydroxymethyl)guanine (DHPG), 5-(2-bromovinyl)2'-deoxyuridine (BVdU), and 5-iodo-2'deoxycytidine (IdCyd) were drastically diminished (between 50-fold and more than 100-fold). This mutation therefore seems to affect the active site of the enzyme which is involved in the catalytic conversion of thymidine and in the binding of the analogues. The above features of HSV-1 R100 seem quite distinct from those of previously described HSV-1 resistant mutants. PMID- 2554612 TI - Analysis of parvovirus infections using strand-specific hybridization probes. AB - The autonomous parvoviruses cause a broad spectrum of acute and chronic infections of animals and man. The discrimination of sites of viral replication from sites of viral sequestration is an important goal in elucidating the pathogenesis of these diseases. It is possible to employ strand-specific RNA hybridization probes in such analyses because a 'plus' sense probe will react with single stranded virion DNA and duplex replicative form DNA, but a 'minus' sense probe will react preferentially with obligate replicative intermediates (duplex replicative form DNA and mRNA). Strand-specific RNA hybridization probes were developed for the Aleutian mink disease parvovirus (ADV) and were used to study acute and chronic infections of mink. Such probes were capable of differentiating replicative intermediates (duplex replicative form DNA and mRNA) from single-stranded virion DNA in Southern blot analysis and in strand-specific in situ hybridization. ADV infection of seronegative newborn mink kits causes an acute, cytopathic infection of type II alveolar cells. Replication in these cells is highly permissive and is characterized by high levels of replicative intermediates and virion DNA. A fatal respiratory distress syndrome and hyaline membrane formation result from impaired surfactant production by the infected type II cells. On the other hand, ADV infection of adult mink is associated with a persistent infection and a disorder of the immune regulation. The target cells for viral replication in adult mink are confined to the lymphoid system and the bone marrow. Replication in these cells, which are probably lymphocytes, is restricted, and characterized by greatly reduced levels of replicative intermediates and virion DNA. It, therefore, seems that disease in the infected adult mink results from a restricted infection by ADV. Large amounts of virion DNA can also be demonstrated in locations where replication cannot be detected and apparently represents sequestration of virion particles by elements of the reticuloendothelial system. Thus, replication and sequestration can, in fact, be distinguished by the strand-specific in situ hybridization. These studies indicate that strand-specific in situ hybridization is a potentially valuable method for studying the pathogenesis of parvovirus infections. PMID- 2554613 TI - Episomal HPV 16 DNA isolated from a cervical carcinoma presents a partial duplication of the early region. AB - An invasive cervical carcinoma was found to harbor an episomal variant of human papillomavirus (HPV) type 16 DNA, with a size of about 10.1 kb. A genomic library of the tumor was constructed in bacteriophage lambda and a recombinant phage clone was isolated by screening with HPV 16 probe. Analysis by restriction mapping and Southern hybridization showed that the isolate contained a 2.2 kb duplication of the early region, which included part of E6, all E7 and part of E1 open reading frames. Possible consequences of this duplication for oncogenesis are discussed. PMID- 2554614 TI - Inhibition of murine coronavirus RNA synthesis by hydroxyguanidine derivatives. AB - A series of hydroxyguanidine derivatives, which are substituted salicylaldehyde Schiff-bases of 1-amino-3- hydroxyguanidine tosylate, were tested for the inhibition of RNA synthesis of mouse hepatitis virus (MHV). It was shown that these compounds could selectively inhibit virus-specific RNA synthesis. Every aspect of viral RNA synthesis, including synthesis of negative-stranded RNA, subgenomic mRNA transcription and genomic RNA replication, was inhibited to roughly the same extent. These compounds are the first known inhibitors of coronaviral RNA synthesis and should prove useful for understanding the mechanism of viral RNA synthesis. PMID- 2554615 TI - Regulatory sequences of SV40 variants isolated from patients with progressive multifocal leukoencephalopathy. AB - Variants of the simian polyomavirus SV40 have been associated in humans with a small number of cases of progressive multifocal leukoencephalopathy (PML). Genomic regulatory regions of two independent isolates, SVPML-1 and -2 [L.P. Weiner et al., N. Engl. J. Med. 286, 385 (1972)], were analyzed to determine if they had acquired any sequences that are present in the genome of the human polyomavirus JC (JCV), the primary causative agent of PML. As compared to SV40 DNA, 83% of SVPML-1 and -2 cloned DNAs contained an apparent deletion of about 30 base pairs in the restriction fragment containing the regulatory region (type alpha); 17% had a deletion of about 60 base pairs (type beta). The regulatory sequences were identical in all four clones representative of SVPML-1 and -2 type alpha grown in human and monkey cells. Thus the primary genomes of two isolates of SV40 from PML are identical in the region that is highly heterogeneous in JCV. The SVPML-alpha-DNAs have a net deletion of 30 base pairs and rearrangements within the transcriptional enhancer. The transcriptional promoter and origin of DNA replication is unaltered from that of SV40. Therefore the human neurotropism of SVPML appears to be a consequence of important rearrangements of SV40 sequences rather than acquisition of JCV-like sequences. PMID- 2554617 TI - [Hydroxyapatite use in dental practice. 1. Use of HA in periodontology]. PMID- 2554616 TI - [The role of GABA-A and GABA-B receptors in the development of amnesia]. AB - The paper deals with analysis of the influence of blockade of separate components of benzodiazepine-GABA-ionophore complex on the recovery of memory trace amnesia under GABA-A and GABA-B receptors activation in the experiments with conditioned reaction of passive avoidance of mice. Activation of GABA-A receptors did not change the behavioural amnesia manifestation at all terms of testing. Activation of GABA-B receptors before learning and amnestic influence caused spontaneous recovery of avoidance reaction. Blockade of chloride channel by picrotoxin and of benzodiazepine receptor by flumazepil restored the reproduction of the memory trace disturbed against the background of GABA-B receptors activation. Systemic flumazepil administration contributed to the memory trace reproduction against the background of GABA-A receptors activation by muscimol in the dose of 0.5 mg/kg. In conditions of amnesia development against the background of muscimol in the dose of 1 mg/kg the blockade of any component of benzodiazepine-GABA ionophore complex was not effective. The obtained data testify that activation of GABA-A and GABA-B receptors changes the amnesia development and correction of amnesia memory trace by the blockade of separate components of benzodiazepine GABA-ionophore complex. PMID- 2554618 TI - [Cool diamond burrs in test conditions]. AB - In almost 500 differentiated measurements of temperature and grinding performance of diamond grinding tools with and without cooling grooves--in conjunction with both a turbine and a high-speed angle piece--under practical conditions, it is demonstrated how many-sided this complex of issues is; it is further demonstrated that it is not possible to generally conclude that all so-called CD grinding tools have better cooling capabilities and/or that they are better suited to prevent excessive heat generation in comparison with grinding tools without cooling grooves. This is equally applicable to grinding performance. PMID- 2554619 TI - [Therapy possibilities in threatened failure of implants]. AB - Parodontological and mucogingival treatment are very successful in case of a prognosed failure of implants. If however they are in vain, the bony defect can be filled up with hydroxylapatit (HA). You must apply HA with great care well aware that there are hardly any long term success described. Future will prove the osseointegration and indulgence of HA in the bone. PMID- 2554620 TI - [Surgery of malignant hepatoma of the cirrhotic liver]. AB - The surgical approach to malignant hepatoma in cirrhotic liver has continued to be an unresolved problem. Early diagnosis, differentiated indication, and surgical techniques need further improvement, before adjuvant therapy can be discussed. All surgeons are called upon to verify experience of the past and to integrate previous findings into a study that should be helpful in overseeing larger numbers of cases. PMID- 2554621 TI - [Somatomedins--insulin-like growth factors]. AB - Growth factors act after specific binding with cell membrane receptors, i.e. these factors mediate mitogenic signals. Insulin-like growth factors (IGF) (syn.: somatomedins) as well as insulin have the same biological activity caused by structural homology. But under normal physiological conditions neither IGF I nor IGF II appear to be involved in the regulation of glucose homeostasis, in contrary to insulin IGFs have mostly mitogenic features. On the other side it is possible that under pathophysiological conditions hypoglycemic effects are caused by an increase of free IGF in the circulation. Insulin acts as a regulating factor in the GF-expression. IGF-I and IGF-II are different peptides especially regarding to their biological role. The synthesis of IGF-I secretion in the liver is dependent on growth hormone (GH). GH is secreted by the pituitary under the influence of the growth-hormone releasing factor (GHF) and is inhibited by somatostatin. In response to GH the liver secretes somatomedin which exerts negative feed back effects on the pituitary and stimulates somatostatin release. The IGF-synthesis is dependent on the human placental lactogen (HPL), i.e. IGF-II is mainly responsible for fetal development the estimation of the production of IGFs by the fetus supports investigations about fetal somatomedins to clear fetal growth retardations and diabetic macrosomia respectively. PMID- 2554622 TI - A bovine herpesvirus (BHV-4) as passenger virus in ethmoidal tumours in Indian cattle. AB - A herpesvirus was isolated from tumours of the ethmoidal mucosa in two of three head of cattle in the State of Kerala, India. The virus designated M40 was cytopathic for a variety of cultured bovine and porcine cells and it did not kill suckling mice or chicken embryos. Sera from tumour-bearing cattle and goats reacted with the M40 virus. Immunofluorescence tests with FITC-conjugated IgG from a bovine monospecific antiserum to bovine herpesvirus 4 (BHV-4) stained the M40 virus specific antigen in infected cells. Experimental infection of goats with the M40 virus did not result in development of tumours. This virus is therefore considered to represent a "passenger" virus. A great similarity was found between restriction patterns of DNAs extracted from M40 virus and the strain 66-P-347, a reference strain of the BHV-4 group. PMID- 2554623 TI - Vaccination of pigs against Aujeszky's disease by the intradermal route using live attenuated and inactivated virus vaccines. AB - A study was undertaken of the protection induced by inactivated and live Aujeszky's disease virus vaccines. The vaccines were administered using a special device which, without the use of a needle, delivered the preparation intradermally. The trials were performed on 88 pigs which were vaccinated at the beginning of the fattening period both in experimental conditions and in pig herds. All the pigs were challenged at the end of the fattening period in isolation units. The results obtained were compared with those obtained using the same vaccines injected intramuscularly. It was shown that vaccination via the intradermal route induced good protection in the vaccinated animals and was similar to that conferred by live virus vaccine injected intramuscularly. The results, with the inactivated virus vaccine, were not so good when it was injected via the intradermal route. Studies with intradermal vaccination showed no local lesion or very small nodules strictly localized to the dermis. The results also confirmed that the effects of challenge exposure depended on the health status of animals prior to infection and show the necessity to use a synthetic value (delta G) to interpret the data and mainly to compare the results objectively. In fattening pigs this vaccination procedure is attractive because (i) less animal constraint is needed than would be for intramuscular injections, (ii) injection can be checked by the presence of a visible papula at the site of inoculation and, (iii) pigs can be vaccinated in the ham while they are feeding. Injection without a needle also contributes to avoiding bacterial contamination under practical farm conditions of vaccination. PMID- 2554624 TI - [The prevalence of antibodies to influenza virus and respiratory coronavirus in fattening pigs in Spain]. AB - The presence of antibodies to two influenza viruses of the type A (H1N1 and H3N2) and to a porcine respiratory coronavirus was investigated in a study lasting a year. 735 blood serum samples were collected from 79 closed pig fattening farms in the province Segovia (Spain). Hemagglutination inhibition was used with influenza viruses. The percentage of positive results was 78.5% and 62.5% respectively for the serotypes H1N1 and H3N2. A clear reduction in the spread of antibodies was observed in the autumn. The ELISA technique was used with the porcine respiratory coronavirus. As antigen we used the antigenically related transmissible porcine gastroenteritis virus. Using this technique 87% of the sera were positive. Some of these sera with representative ELISA values were confirmed by means of serum neutralisation and radioimmune precipitation of the viral proteins. The incidence of these antibodies remained unchanged over the whole year of the investigation. PMID- 2554625 TI - [The relation of seasonal manifestations of herpes zoster and the seasonal periodicity of normal humoral immunity]. AB - 29 practically healthy persons and 97 herpetic ganglionitis patients were examined at different periods during 1986-1987. The seasonal changes of humoral immunity both in clinically healthy persons and in herpetic ganglionitis patients were established. The conclusion was made that clinical manifestations of herpetic ganglionitis are linked with a decrease in the level of IgG ensuring natural resistance to chickenpox virus. PMID- 2554626 TI - [Delayed hypersensitivity to the antigens of the herpes simplex virus and herpetic resistance induced in mice by the administration of xenogeneic lymphocytic ultrafiltrates]. AB - Experiments on mice have shown that ultrafiltrates, prepared from lymphocytes obtained from the spleen of horses immunized with herpes vaccine and from human tonsils, contain herpes-specific transfer factor inducing delayed hypersensitivity. The antiherpetic resistance of mice has been found to achieve its maximum if herpes simplex antigens are introduced after the injection of the preparation of transfer factor. PMID- 2554627 TI - [Computed tomographic diagnosis of paraganglioma of the base of the skull]. AB - On the basis of examination of 30 patients with the diagnosis of paragangliomas of the base of the skull the following main computed tomographic signs of this group of tumors were revealed: destruction of bone structures of the base of the posterior and middle cranial fossae, erosion and widening of the jugular foramen; diminished pneumatization of the air cells of the mastoid process; extracranial simultaneous with intracranial spreading of the tumor. A classification of paragangliomas is suggested. PMID- 2554628 TI - [Cyclic adenosine monophosphate in the cerebrospinal fluid of patients with brain trauma and diseases]. AB - The authors studied adenosine 3,5-cyclic monophosphate (cAMP) in the cerebrospinal fluid (csf) of patients with craniocerebral trauma (CCT), ischemic stroke, and brain tumors. A statistically significant increase of cAMP in csf of patients with moderate and severe CCT was revealed. In unfavourable outcomes of severe CCT the increase of cAMP continued to the time of death. The authors suggest using this index as a diagnostic criterion of the depth of the metabolic disorders in the brain tissue, as a prognostic criterion, and for purposes of differential diagnosis. The possible mechanism of lipid peroxidation activation in CCT is discussed. PMID- 2554629 TI - [Serum level of neuron-specific enolase: value as tumor marker of microcellular bronchial cancer]. AB - This study analyses retrospectively the plasma level of neuron-specific enolase (NSE) performed in 236 patients presenting for the diagnosis of a bronchopulmonary disease. Taking a cut off on 15 micrograms/L, 61 (64%) of the 95 patients sharing a small cell lung cancer (SCLC) (45% of those with a limited disease and 90% with an extensive disease) but 8 (13%) of 60 with a non small cell lung cancer patients and 0% of those with a benign bronchopulmonary disease showed an abnormal value of NSE. Therefore in this group of patients, the diagnosis of SCLC could be assessed with a sensitivity of 64% and a specificity of 94% in patients with an NSE plasma level above 15 micrograms/L. In addition, the interest of sequential dosages of NSE for the monitoring of SCLC patients is also stressed. PMID- 2554630 TI - Effect of desferrioxamine B on hemolysis in glucose-6-phosphate dehydrogenase deficiency. AB - Twenty-four infants and children suffering from glucose-6-phosphate dehydrogenase (G6PD) deficiency during hemolytic crisis were included in this study. Their ages ranged between 3 and 36 months with a median of 10 months. 22 were males and 2 were females. Fourteen out of them received a single bolus dose of desferrioxamine B 500 mg intravenously followed by packed red cell transfusion, while the remaining 10 cases were only transfused. Sequential estimation of hemoglobin level, reticulocytic count and hemoglobinuria was done before treatment, 3, 24, 48 and 72 h thereafter. The hemoglobin level was higher in the desferrioxamine B group. The degree of increase was statistically significant at 48 and 72 h (p less than 0.01). Hemoglobinuria stopped in 78.5% in the first group and only in 30% of the second group at 72 h. It was concluded that desferrioxamine B is helpful in shortening the course of hemolytic crisis in G6PD deficient patients. It could be used as an adjuvant to packed red cell transfusion. PMID- 2554631 TI - Investigations on beta-glucuronidase-positive erythrocytic inclusions. AB - Novel erythrocytic inclusion bodies, characterized by strong beta-glucuronidase activity, are described. Their physicochemical properties differ substantially from those of known inclusion bodies and they are mainly observed in patients with liver damage. Since these inclusions occur almost exclusively in erythrocytes, the intraerythrocytic beta-glucuronidase must originate from an extraerythrocytic source, perhaps entering erythrocytes via a receptor-mediated endocytotic pathway. PMID- 2554632 TI - Infection of hematopoietic and stromal cells in human continuous bone marrow cultures by a retroviral vector containing the neomycin resistance gene. AB - Stability and expression of the bacterial neomycin resistance gene (neor) transferred to human continuous marrow cultures by a retroviral vector [pZIP NeoSV(X)] was evaluated over 4 weeks. Following infection of long-term human marrow cultures with pZIP-NeoSV(X), 10-15% of the stromal cells demonstrated high replating efficiency in a dose of the neomycin analogue G418 that was toxic to stromal cells from uninfected cultures. In contrast, G418 resistance was detected in less than or equal to 1% of GM-CFUc and CFU-GEMM derived from the same virus infected compared to control cultures. Infection of human CFU-GEMM enriched 100 X by monoclonal antibody selection with pZIP-NeoSV(X) did not increase the percentage of neor progenitors. Marrow cells from cultures infected with pZIP NeoSV(X) and a replication competent amphotropic virus transferred the vector and G418 resistance to HeLa cells at a frequency of 1/10(5) for nonadherent and 1/10(4) for adherent cells. Two established human hematopoietic (HL60 and K562) and one stromal cell line (KM101) stably expressed the neor gene. Thus, a higher efficiency of infection and expression of a gene transferred by pZIP-NeoSV(X) to permanent human hematopoietic tumor cell lines and fresh marrow stromal cells contrasts with a lower level of expression in fresh CSF-dependent human hematopoietic stem cells. PMID- 2554633 TI - Post-irradiation lesions of the caudal roots. AB - The article reports on 3 patients suffering from muscular atrophy after radiotherapy of the para-aortal lymph nodes for malignant testicular tumor without any sensory, bladder, or bowel disturbances. By neurophysiological examination, a lesion of the lumbal plexus and the peripheral nerves of the lower extremities were excluded. On EMG-examination there were no giant motor unit potentials, as they can be found in anterior horn cell lesions. Though there were no sensory deficits, a distinct prolongation of latencies and reduction of amplitudes could be found for lumbar dermatomal somatosensory evoked potentials (SSEP) and those after stimulation of some peripheral nerves of the lower extremities. PMID- 2554634 TI - Polyneuropathy: an unusual extraintestinal manifestation of Crohn's disease. AB - Among the extraintestinal complications of Crohn's disease, neuropathy seems to be rare, and is only exceptionally reported in the literature. The authors present the report of a patient followed over a 12-year period who developed polyneuropathy with a parallel course to Crohn's disease. There was no evidence of any other underlying condition. On one occasion, the patient developed a cutaneous vasculitis with fibrinoid deposits in the vessels. The search for circulating immune complexes was positive. The authors demonstrated increased intestinal permeability using the 51chromium-labeled ethylenediaminetetra-acetate (51Cr-EDTA) test. Ultrastructural study of a biopsy of the superficial peroneal nerve disclosed a marked denervation process. Therapeutic plasmapheresis were performed, improving the neurologic symptoms. Vasculitis with circulating immune complexes is suggested as a possible mechanism in view of the occurrence of cutaneous vasculitis, increased intestinal permeability, and plasma exchange efficiency. PMID- 2554635 TI - Sporadic motor neuron disease with Lewy body-like hyaline inclusions. AB - Lewy body-like hyaline inclusions were immunocytochemically and electron microscopically investigated in a patient with sporadic motor neuron disease. The hyaline inclusions were chiefly observed within the perikarya of both normal looking and chromatolytic anterior horn cells in the lumbar spinal cord, but some were detected in the axons and dendrites. Usually, a single inclusion was found in the perikaryon, but in rare cases two or more were observed. Immunocytochemically, these inclusions were intensely immunostained with anti ubiquitin antibody. Ultrastructurally, the hyaline inclusions were chiefly composed of randomly arranged linear structures associated with ribosome-like granules, varying from compactly arranged linear densities to more loosely packed ones. They contained scattered vesicles of various sizes and occasionally a focal accumulation of randomly arranged 10-nm neurofilaments or 13-25-nm filamentous structures. PMID- 2554636 TI - Ultrastructure of capillary walls in human brain tumors. AB - Changes in capillary walls between human glial, non-glial and metastatic brain tumors were studied with conventional ultrathin section and freeze-fracture replica techniques. The following results were obtained. (1) In glial tumors, ultrathin section studies showed cell junctions of the capillaries were either short or elongate. Moreover, endothelial hyperplasia, surface infolding of endothelial cells, irregularity of the basal lamina and a large extravascular space were observed. Freeze-fracture replicas of capillary endothelium showed tight junctions as two to seven strands. In addition, pinocytotic vesicles had increased markedly and were an average of 25 per microns 2. Both ultrathin and freeze fracture studies showed that, in contrast to malignant gliomas, there were only slight changes in benign astrocytomas. (2) In non-glial tumors, ultrathin sections showed surface infoldings, increased vesicles, many fenestrations of endothelial cells, irregularity of basal lamina and enlarged perivascular space. Freeze-fracture replicas of vascular endothelium, showed that the average number of pinocytotic vesicles and fenestrations were 25 and 22 per microns 2, respectively. Moreover, the tight junction was composed of one or two strands which appeared to be a discontinuous array of particles. (3) In metastatic brain tumors, ultrathin studies showed capillary endothelia were proliferated, had marked infolding, and showed an increased number of pinocytotic vesicles and many fenestrations. Moreover, short and elongate intercellular junctions were presented but no open junction was detected. Finally the basal lamina lost its three-layered appearance and was irregular in width. Freeze-fracture replicas showed pinocytotic vesicles had increased and were 24 per microns 2 on average in four cases, but fenestrations and tight junctions could not be detected. The most fundamental feature of vessels in these three different kinds of tumors was whether they were fenestrated or not. Glial tumors were non-fenestrated, whereas non-glial and metastatic tumors were fenestrated. PMID- 2554637 TI - Festschrift in honour of Claes Dohlman. PMID- 2554638 TI - Surface cell morphology of the anesthetic human cornea: a color specular microscopic study. AB - The technique of color specular microscopy has been utilized to study the surface cell morphology of the human cornea in 6 subjects with profound corneal anesthesia from diverse causes. In an analysis of cell area, perimeter, and shape, anesthetic corneas showed a statistically significant increase in cell size and perimeter. In addition, there was a loss of cellular outlines and overall increase in rose bengal staining of cells. We believe that these cellular microscopic changes reflect profound changes in epithelial cell turnover related to a decrease in neurohumoral transmitters known to be necessary for maintenance of normal corneal morphology and function. PMID- 2554639 TI - Keratopathy of the rabbit cornea following complete eyelid closure. AB - The corneal response to a complete tarsorrhaphy was studied in 46 rabbits for time periods up to 21 days. During the first 7 days of complete eyelid closure corneal thickness increased up to 18.4%, glycogen decreased 32.2%, and lactate rose 27.2%. A steep increase in corneal thickness to 89.8% appeared after 14 days, followed by decreasing values at 21 days, which was concomitant with the formation of extensive vascularized pannus. These results confirm previous findings that the partial pressure of O2 under closed eye conditions is substantially below physiologic requirements, affects endothelial pump capability, and results in major corneal swelling. When silicone contact lenses were fitted immediately before eyelid closure, corneal swelling 2 days after lid closure did not differ from that in eyes without lenses. PMID- 2554640 TI - Johan Lorenz Odhelius and cataract extraction in Sweden. AB - In 1775, Johan Lorenz Odhelius presented a lecture to the Royal Academy of Science at Stockholm describing his preferred operation for cataract. Odhelius generally endorsed the technique described by Daviel (1753) 22 years earlier, but included additional observations and modifications based on his own experience. This address was published in Swedish (Odhelius 1775) in the same year and is one of the earliest ophthalmological monographs by a Swedish author. Odhelius' publication is now translated into English and put into its historical perspective. PMID- 2554642 TI - Extraneuronal herpetic latency: animal and human corneal studies. AB - HSV DNA has been previously detected by both in situ and dot blot hybridization in neuronal tissues latently infected with herpes simplex virus (HSV), but not in extraneuronal tissues. The present study, using dot blot hybridization with a cloned full-length HSV DNA probe and subtractive hybridization assays for detecting HSV RNA, reveals both the presence and activity of the HSV genome in 100% of HSV latently infected rabbit corneas tested. Studies on human herpetic corneas taken at keratoplasty using slot blot hybridization with a cloned full length HSV DNA probe demonstrated positive binding (hybridization) to the probe in 50% of samples tested but no binding to normal human control DNA. These studies confirm earlier, less sensitive virus recovery assays and implicate the cornea as an extraneuronal site of HSV latency and reactivation. PMID- 2554641 TI - The molecular basis of neurotrophic keratitis. AB - Endogenous proliferation of corneal epithelial cells is regulated by a bidirectional control process characterized by an adrenergic, cAMP-dependent 'off', and a cholinergic, muscarinic cGMP-dependent 'on' response. The adrenergic receptor(s) are located in the plasma membrane (microsomal fraction), whereas the novel feature of the system is a cholinergic receptor specific for acetylcholine (ACH) located in the nuclear membrane. Exogenous substances which raise intracellular cAMP levels such as isoproterenol or PGE1, shut off epithelial mitosis: and, carbamylcholine or ACH raise intranuclear cGMP levels and increase mitosis by specific, regulatory stimulation of RNA-polymerase II activity. We believe that this regulatory system explains the transitory mitotic suppression induced by superficial corneal wounding (interruption of adrenergic fibres, chalone-effect); and the marked, permanent depression of epithelial mitosis associated with decreased intracellular ACH levels which are produced by total corneal denervation, and which results in neurotrophic keratitis. PMID- 2554643 TI - Evidence for the potential influence of cyclic nucleotides on maintenance of or reactivation from latency of herpes simplex virus in trigeminal ganglionic neurons. AB - Herpes simplex virus infections are epidemic throughout developed countries, and recurrent herpes simplex keratitis is the most common cause of corneal blindness in these countries, NIH (1973). No available antiviral agent is capable of eradicating the state of viral ganglionic latency, and hence no effective treatment currently exists for prevention of viral re-activation from latency, with resultant recurrent infectious viral clinical manifestations. Putative triggers of re-activation include stress, sunburn, menses, trauma, and fever. These 'triggers' seem to share at least one common characteristic: the potential ability to influence intracellular cyclic nucleotide levels through the action of such first order messengers as catacolamines (stress, trauma) and/or arachadonic acid metabolites (sunburn, fever, trauma, and menses). We exploited an in vitro model of HSV ganglionic latency, and developed a model of in vitro organ culture ganglionic viral reactivation from latency. We then examined the effect of a variety of agents on this model. We found that agents which have been shown to elevate cyclic AMP levels consistently produce increased viral shedding (compared to control, spontaneous reactivation rate) in our model of viral reactivation from latency. In contrast, agents which have been shown to depress c-AMP levels and/or to elevate c-GMP levels inhibit viral reactivation from latency in this in vitro model. We conclude that intracellular cyclic nucleotide levels may influence events which control herpes simplex genome transcription. PMID- 2554644 TI - Ocular anaphylaxis induced in the rat by topical application of compound 48/80. Dose response and time course study. AB - In the present study we sought to develop a model of ocular anaphylaxis based on the topical application of compound 48/80 to the surface of the rat eye. Doses ranging from 50 to 1000 micrograms were found to produce graded edema of the conjunctiva and swelling of the lid. On histologic examination, 50 microns compound 48/80 produced no changes distinguishable from those in PBS-treated controls, 150 microns produced mild alterations, and 250, 500, and 1000 micrograms compound 48/80 produced a marked increase in degranulated mast cells and a mild influx of neutrophils. The time course of the response to 250 micrograms and 1000 micrograms of compound 48/80 was evaluated over a 72-h period. Both doses elicited epithelial damage. A mild reduction in the number of mast cell was seen at 6 h in rats receiving 250 or 1000 micrograms. The reduction persisted to 72 h in rats receiving 1000 micrograms. The number of neutrophils was increased at 1 and 6 h in eyes treated with 250 micrograms and at 1, 6, and 24 h in eyes treated with 1000 micrograms compound 48/80. The clinical and histologic changes induced by application of 250 micrograms compound 48/80 resemble those seen in patients with allergic conjunctivitis suggesting that a model of ocular anaphylaxis based on the topical application compound 48/80 will be clinically relevant and experimentally practical. PMID- 2554646 TI - Development of the gas-permeable impression-moulded scleral contact lens. A preliminary report. AB - Although superceded by corneal hard and soft contact lenses for the correction of simple refractive error, the scleral lens still has numerous medical applications in modern ophthalmological practice. However, a major problem with scleral lenses has been the limited wearing time due to corneal occlusion and consequent hypoxia. We present two alternative methods for the production of moulded gas permeable scleral contact lenses. PMID- 2554645 TI - Time course of human conjunctival mast cell degranulation in response to compound 48/80. AB - Compound 48/80, a non-immunogenic mast cell degranulatory agent, is known to produce the signs and symptoms of ocular allergy. Maximal mast cell degranulation of human conjunctiva occurred within the first hour after stimulation by a single topical dose of compound 48/80 (20 microliters, 7.5 mg/ml). The average percentage of fully degranulated mast cells in treated specimens (n = 9) was 31% (range 5-60%) versus 6% (range 0-20%) in control specimens (n = 5). Exact correlates of representative granulated, partially degranulated, and fully degranulated mast cells were determined by light and transmission electron microscopy. PMID- 2554647 TI - The optical quality of the peripheral cornea. AB - The cornea can be considered to be an organ divided into a central and peripheral zone. Although the central cornea is responsible for producing a sharp retinal image, our experiments suggest that areas of isolated peripheral cornea can produce retinal images compatible to 20/30 visual acuity. PMID- 2554648 TI - Long-term survival of large diameter penetrating keratoplasties for keratoconus and pellucid marginal degeneration. PMID- 2554649 TI - Lectin receptors of normal and dystrophic corneas. AB - In recent years, our laboratory has used lectins to detect carbohydrate-related abnormalities in dystrophic corneas and species-specific differences in the carbohydrates of normal corneas. These studies have: 1) demonstrated that abnormal glycoconjugates are present in corneas with macular and lattice dystrophy, 2) provided additional means of differentiating corneas with macular, lattice and granular dystrophy, 3) demonstrated that distinct species-specific differences are present in the glycoconjugates of normal corneas and 4) provided guidelines for the selection of an animal model for the study of corneal glycoconjugates. PMID- 2554650 TI - Keratoconus associated with Terrien's marginal degeneration. A clinical and ultrastructural study. AB - A 59-year-old man presented with Terrien's marginal degeneration associated with keratoconus of his left eye. Penetrating keratoplasty of the left eye permitted an evaluation under optical microscopy, as well as transmission and scanning electron microscopy. An anatomical-pathological evaluation confirmed the co existence of a central lesion due to keratoconus and a peripheral lesion due to Terrien's marginal degeneration. An absence of the knowledge of the etiology of either affliction does not allow one to define whether there exists one or two active degenerative processes. PMID- 2554651 TI - Late corneal opacities in the syndrome of interstitial keratitis and vestibulo auditory symptoms. AB - Progressive clouding of the cornea may be a delayed complication of the interstitial keratitis-vestibuloauditory syndrome. Two illustrative cases are presented. The first case illustrates progressive opacification about aberrant vessels in the deep stroma presenting a characteristic dendritiform pattern. The second case shows that the opacification is due, in this case at least, to formation of connective tissue with inclusion of lipid crystals and fat along with the blood vessels. Descemet's membrane is also thickened several fold. PMID- 2554652 TI - Congenital corneal keloid. AB - The clinical course and pathological findings in an infant with a congenital corneal opacity, aniridia, and cataract of the right eye and anophthalmia of the left orbit are reviewed. Although the lesion was thought to represent a dermoid pre-operatively, the keratoplasty specimen revealed disorganized and vascularized connective tissue, with thickened, keratinized epithelium. Bowman's and Descemet's membranes as well as the endothelium were absent. These histopathologic findings were felt to represent a congenital corneal keloid. The co-existence of aniridia in the involved eye and anophthalmia of the contralateral socket are hypothesized to be evidence that a primary ocular developmental disorder, rather than a reparative process, resulted in the corneal alteration. The difficulties of penetrating keratoplasty in lesions of this sort are examined. PMID- 2554653 TI - The effect of corneal grafting on vision in bilateral amblyopia. AB - Two children with bilateral congenital corneal opacities (sclerocornea) received unilateral corneal transplants at the ages of 4 1/2 and 16 years, respectively. Both developed reading vision and reduced nystagmus excursions. PMID- 2554654 TI - The effect of standardized keratoplasty technique on IOL power calculation for the triple procedure. AB - Twenty-one consecutive triple procedures (keratoplasty, cataract extraction, lens implantation) performed by one surgeon using identical suturing technique, donor size, and donor/recipient size disparity were analyzed for visual outcome and refractive error. Ninety-five percent of all grafts were clear with an average follow-up of 11.8 months. Of patients with good preoperative visual potential, 84% achieved 20/40 or better visual acuity, and the majority of these patients obtained 20/40 acuity within 6 months of surgery. Sixty-three percent of eyes with 20/40 or better acuity had refractions within +/- 2 diopters of the predicted post-operative refraction. The most recent 14 eyes in this series had IOL power calculations performed utilizing the SRK regression formula and 43.00 K's (surgeon's average post-keratoplasty keratometry values). Within this group, 79% achieved 20/40 or better vision. Eighty-two percent of these eyes had refractions within +/- 2 diopters, and 100% were within +/- 3 diopters of the predicted value. These findings demonstrate that a single surgeon using standardized keratoplasty can achieve good refractive results in the triple procedure. PMID- 2554655 TI - Refractive surgery: through the looking glass. AB - Historically, surgical procedures designed to change the refractive power of the cornea have been controversial at their inception, gaining acceptance only with time and study. From intraocular lenses to corneal remodelling with the excimer laser, and numerous procedures in between, including radial keratotomy, epikeratophakia, incisional keratotomy for astigmatism, and hydrogel and polysulfone implants, ophthalmologists have balanced the risks and benefits for their patients to produce the maximum benefit at the least cost. The present status and future potential of each of these are assessed as they appear today. There is no question that progress will continue, and that our patients will be the beneficiaries of our ongoing interest in improving older procedures and developing new techniques for the correction of refractive errors. PMID- 2554656 TI - Epikeratoplasty for adult and pediatric aphakia, myopia, and keratoconus: the Massachusetts Eye and Ear Infirmary experience. AB - Between February, 1985 and February, 1987, 49 eyes of 47 patients underwent epikeratoplasty for one of the following indications: adult aphakia (21 eyes; 20 patients), pediatric aphakia (10 eyes; 10 patients), myopia (7 eyes; 6 patients), and keratoconus (11 eyes; 11 patients). Follow-up at least 12 months (average = 18 months) is available for all 49 eyes. Of 49 eyes, 44 (90%) were anatomically successful with an intact, clear lenticule at least 12 months post-operatively, with no significant difference between the four groups (adult aphakia = 90%, pediatric aphakia = 90%, myopia = 86%, and keratoconus = 91%). The functional success rate, however, was 91% for keratoconus, 81% for adult aphakia, 80% for pediatric aphakia, and only 43% for myopia. We conclude that epikeratoplasty can be an effective tool in the treatment of carefully selected patients with adult or pediatric aphakia and keratoconus, but has only limited applicability in the management of myopia. PMID- 2554657 TI - Surgical procedures in the treatment of most severe eye burns. Revival of the artificial epithelium. AB - Report of some clinical observations in 16 patients with severe and most severe burns of 19 eyes. Major problems were delayed regeneration of the epithelium, extensive ulceration of the sclera near the limbus, and subsequent corneal ulceration. The therapy with anti-inflammatory drugs, macromolecular hyaluronic acid, fibronectin, epidermal growth factor, free transplants of oral mucosa and conjunctiva was sometimes helpful, but could not induce stable epithelial regeneration. The major problem remained the inability of the epithelium to regenerate on the cornea. Therefore, an artificial epithelium was applied in many cases. When healthy conjunctiva touched closely the artificial epithelium at the limbus, it remained tightly attached to the corneal stroma, preserved it's structure and even prevented corneal vascularisation for many months. However, when corneo-scleral ulceration was present, the artificial epithelium failed to protect the corneal stroma from secondary ulceration and sloughed off. Therefore, new surgical procedures, a conjunctiva-Tenon flap and a Tenon plasty were introduced to cover and to cure corneoscleral ulceration. These operations formed a new limbus, attached tightly to the rim of the artificial epithelium and prevented secondary ulceration underneath. Then under the protection of the artificial epithelium and the Tenon plasty the eye could be treated topically with corticosteroids for a long time. When the inflammatory response to the burn was healed, the natural epithelium recovered and the eye became ready for successful keratoplasty. PMID- 2554658 TI - The pathogenesis of corneal epithelial defects. PMID- 2554659 TI - Distribution of collagenase and cell types in sterile ulceration of human corneal grafts. AB - Extensive clinical and experimental evidence has suggested a role for corneal epithelium, keratocytes, and acute inflammatory cells in sterile corneal ulceration, although the precise role of these cell types in stromal matrix degradation has not been elucidated. We studied two corneal buttons and two whole corneas from 4 patients with corneal grafts and sterile stromal ulceration. Each cornea was divided into several sections and examined morphologically, while adjacent sections were placed in tissue culture and assayed for collagenase activity against type I collagen. In each case, collagenase activity was present in tissue from ulcerating and nonulcerating areas and, in most regions assayed, exceeded collagenase activity in normal control peripheral donor corneas. Collagenase levels were not always greatest in areas of most advanced ulceration. Although polymorphonuclear leukocytes were extensively present histologically in two cases, one case had a mixed inflammatory cell population at the site of ulceration, and another had only keratocytes (fibroblasts) present in the entire cornea. We conclude that regional differences in collagen degradation are due not only to the presence of collagenase, but also to cellular and humoral activators and inhibitors that determine collagenolytic activity. PMID- 2554660 TI - Repair of the corneal epithelial adhesion structures following keratectomy wounds in diabetic rabbits. AB - In corneal tissue from a diabetic patient 15 months after delayed epithelial healing following vitrectomy surgery, electron microscopy revealed a discontinuous basement membrane; immunohistochemically, fibronectin was present in the basement membrane zone. To determine whether alterations in repair of the basement membrane, anchoring fibrils and hemidesmosomes occur in wounds in diabetics, we studied the pattern of histochemical localization of bullous pemphigoid antigen (the intracellular hemidesmosome plaque component), laminin and type VII collagen (the anchoring fibril collagen) in alloxan-induced diabetic rabbits following superficial keratectomy. Ultrastructural studies of basal laminae and hemidesmosomes also were performed. Epithelial wound closure was complete by 66-72 h after 7-mm superficial keratectomies. Type VII collagen localized at the base of the epithelium at the wound periphery by 66 h, appearing as a beaded line underlying the epithelium; at 1 week, as in control rabbits, the localization zone extended across the wound bed. Polyclonal antibodies to laminin and bullous pemphigoid antigen showed similar localization. Small segments of basal laminae were noted by electron microscopy of the wound periphery as early as 66 h post-keratectomy. By 2 weeks, an increase in hemidesmonsomes associated with segments of discontinuous basal lamina was apparent. No obvious differences in the time or pattern of re-appearance of the adhesion complex in keratectomy wounds could be discerned between normoglycemic and hyperglycemic rabbits. Thus, healing defects in diabetics may be due to factors other than reassembly of adhesion structures. PMID- 2554661 TI - The role of the limbus in ocular surface maintenance and repair. AB - Since Claes Dohlman's original treatise on the corneal epithelium, a great deal has been learned about the biology of that layer. Its maintenance depends in part on centripetal movement of cells from the periphery toward the center. To some degree the supply of these peripheral cells appears to be dependent on the proliferation of cells at the limbus, from so-called 'stem cells'. Studies of stem cell replication and differentiation of their progeny will lead undoubtedly to new insights into the the mechanisms responsible for many of the chronic abnormalities of the central corneal surface. PMID- 2554662 TI - Natural history of disease in a rabbit model for keratoconjunctivitis sicca. AB - We have continued our study of the tear film and ocular surface in our full KCS (keratoconjunctivitis sicca) rabbit model up to 52 weeks post-operatively. Tear film osmolarity remains elevated, conjunctival goblet cell density remains decreased, and the conjunctival epithelium remains abnormal. Corneal epithelial glycogen levels decreased progressively, and at 44 weeks post-operatively rabbits developed abnormal rose Bengal staining of the affected cornea that was shown to be associated with morphologic abnormalities at 52 weeks. Rabbits began rubbing the affected eye after the development of corneal staining. Our full KCS rabbit model demonstrates the features of the human disease. PMID- 2554663 TI - Small liver masses in cirrhotic patients. A pathological clue for the morphogenesis of human hepatocellular carcinoma. AB - Eleven small primary liver lesions detected in four cirrhotic patients are reported in detail. Some lesions showed the histopathological features of macroregenerative nodules (MRN) containing foci of cellular and structural atypia or of hepatocellular carcinoma (HCC). Other lesions showed the histological abnormalities of the early developmental stage of HCC without a background of MRN. Finally one lesion is described with an equivocal background between benign and malignant, containing a focus of carcinoma. All lesions presented suggest the presence of different pathways in the morphogenesis of human HCC. PMID- 2554664 TI - A unique verrucous anogenital tumor associated with type 6b-related human papillomavirus. AB - Multiple dark brown papillomatous tumors, showing some histological features of verrucous carcinoma or giant condyloma, developed mainly in the anogenital region of a Japanese woman. The tumors first appeared when she was 51 years old and annoyed her for over 20 years with several recurrences without any frank malignant transformation, after surgery. Immunohistochemically, papillomavirus genus-specific antigen was demonstrated only in small foci of the lesions resected at first operation. Southern blot analysis revealed human papillomavirus type 6b-related DNA in a surgically resected specimen. The possible role of the human papillomavirus in the genesis of this unique tumor is discussed. PMID- 2554665 TI - [P-8502--a new mu selective opioid receptor ligand]. AB - The analgesic potencies of 3-[beta-(p-amino)-phenethyl)-9 beta-methoxy-9 alpha-(m methoxyphenyl)-3- azabicyclo [3,3,1] nonane (P-8502) and 3-[beta-(p-monoester fumarylamido)-phenethyl]-9 beta-methoxy-9 alpha-(m-methoxyphenyl)-3-azabicyclo [3,3,1] nonane (P-8511) were examined. The analgesic ED50 of P-8502 and P-8511 were 55 and 200 micrograms/kg (mice, ip, hot plate), and 30 and 95 micrograms/kg (rat, sc, tail flick), respectively. The duration of the analgesic action of P 8511 (about 4 h) was longer than that of P-8502 (about 1.5 h, rat, sc, tail flick). Binding assay showed that P-8502 had a high ratio of delta/mu, kappa/mu: IC50 (DPDPE)/IC50 (DAGO) = 399; IC50(DAD-LE)/IC50 (DAGO) = 1498; IC50 (kappa)/IC50 (DAGO) = 159. In conclusion, P-8502 appears to be a new mu selective opioid receptor ligand, whereas P-8511 has no such selectivity. PMID- 2554667 TI - [Hypothermic effect of intracerebroventricular injections of taurine on endotoxin induced fever in rabbits]. AB - Taurine (Tau) was infused by intracerebroventricular (icv) injection 20 min after iv endotoxin (ET). Cerebrospinal fluid was taken from the posterior cisterns at 0, 60, 210 and 390 min after Tau infusion. The concentrations of cAMP and PGE2 in the samples were determined by radioimmunoassay. In the control groups, an equivalent isotonic saline (NS) (non-pyrogenic) instead of Tau or ET was used respectively. The rectal and ear skin temperatures of the rabbits in a non fasting state were recorded automatically. Tau icv 1.0 mg/50 microliters, followed by the slow infusion of Tau 0.06-0.22 mg/(kg.min) for about 20 min into rabbits, caused sedation and peripheral vasodilation (rise in ear skin temperature), and initially blocked the rise in rectal temperature induced by ET (iv, 1 micrograms/kg). The results for the control groups were significantly different from those of the ET + Tau group. In the ET + NS group, the fluctuations in concentrations of PGE2 and cAMP paralleled the change in rectal temperature, but in the ET + Tau group, the changes in PGE2 and cAMP concentrations were similar to those of the ET + NS group, even though the fever was initially inhibited. There were no changes in the concentrations of these mediators in the NS + Tau group. PMID- 2554666 TI - [Protective effects of m-nisoldipine and nisoldipine on myocardial damage in working rabbit hearts after ischemia-reperfusion]. AB - m-Nisoldipine 4, 8, 16 nmol/L, nisoldipine 1, 4 nmol/L and nifedipine 4, 8, 16, 50 nmol/L enhanced the recoveries of functional parameters of working rabbit hearts after ischemia-reperfusion, as well as prevented the development of contracture and the release of CPK from the reperfused hearts. m-Nisoldipine 8 nmol/L, nisoldipine 1 nmol/L and nifedipine 8 nmol/L attenuated the reduction of myocardial Na+-K+-ATPase and 5'-nucleotidase activity induced by ischemia reperfusion. The breakdown of membrane phospholipids and elevation of the myocardial Ca2+-ATPase activity and the free fatty acids level were also prevented. PMID- 2554668 TI - Inhibitory effect of disodium cromproxate on superoxide anion (O2-) generation and membrane potential changes in stimulated neutrophils. AB - Disodium cromproxate is an antiallergic agent. This drug (0.5-2 mmol/L) inhibited O2- production by neutrophils induced by FMLP and PMA. However, the inhibition of FMLP-induced O2- generation was more pronounced than that induced by PMA. Disodium cromproxate also counteracted the changes in membrane potential in neutrophils induced by either FMLP or PMA. The actions of disodium cromproxate differed from those of propranolol, as propranolol had no antagonistic action on membrane potential changes induced by FMLP and PMA. PMID- 2554669 TI - [Antiviral effect of mangiferin and isomangiferin on herpes simplex virus]. AB - Using tissue culture techniques the present study assured us of the merits of mangiferin and isomangiferin in the antiviral action against HSV-1. Utilizing 4 main patterns for evaluating drug effectiveness (ie intratube drug-on-virus direct action, simultaneous addition of drug-virus-inoculum to cell bottle, virus inoculation preceding drug addition, and drug addition followed by virus inoculation), it was readily found by logarithm determination of HSV-I inhibition that isomangiferin was superior to such control drugs as acyclovir, idoxuridine, and cyclocytidine in logarithm by 0.27-0.50, and that mangiferin was lower than isomangiferin in logarithm by 0.53. The average plaque reduction rates of mangiferin and isomangiferin were 69.5% and 56.8%, respectively. All in all, the antiviral effect of mangiferin and isomangiferin was attributed presumably to their capability to inhibit virus replication within cells. PMID- 2554670 TI - [Effects of an endogenous GABA receptor binding inhibitor on rat blood pressure]. AB - The GABA receptor agonists GABA (400 micrograms icv) and muscimol (1 microgram icv) induced hypotension in urethane-anesthetized rats, while the GABA receptor antagonist bicuculline (2 micrograms icv) elicited hypertension. An endogenous GABA receptor binding inhibitor (1 mg), prepared from bovine cerebellum, showed bicuculline-like hypertensive action in a dose-dependent manner. It was antagonized by icv muscimol, but not by the alpha 2-adrenoceptor agonist clonidine. These in vivo results agree quite well with our previous in vitro receptor binding assay experiments. PMID- 2554671 TI - [Effect of tolmetin on responsiveness of isolated guinea pig tracheal smooth muscles]. AB - Tolmetin directly relaxed the isolated guinea pig tracheal smooth muscles, increased the muscle responsiveness to histamine or carbachol and enhanced the tracheal Schultz-Dale reaction. Prostaglandin E2 synthesized by cyclooxygenase was transformed into prostaglandin B2 (PGB2) with alkali, PGB2 and the lipoxygenase metabolite leukotriene B4 (LTB4) were measured by high-performance liquid chromatography, whereas slow-reacting substance of allergy (SRS-A) was determined by bioassay. Tolmetin showed inhibitory effects on prostaglandin E2 synthesis, with the IC50 being 30 mumol/L, but no effect on LTB4 synthesis (up to 100 mumol/L). SRS-A release was potentiated by tolmetin at 10 and 50 mumol/L. The effects of tolmetin on arachidonic acid metabolism may be responsible for its effects on tracheal smooth muscles. It is suggested that asthma attacks may be induced or enhanced by tolmetin in patients with asthma. PMID- 2554672 TI - [Irreversible action of the opioid agonist alpha-CAM and its reaction with SH groups at opioid receptor binding sites]. AB - 7 alpha-Bis (beta-chloroethyl)amino-methyl-6,14-endoethenotetrahydrooripavine (alpha-CAM) was found to bind to opioid receptors irreversibly and react directly with sulfhydryl (SH) groups in P2 preparations of rat brain. The P2 preparations were pretreated as follows: protection of the SH groups at the opioid receptor binding sites by morphine or etorphine, and inactivation of the SH groups outside the binding sites by N-ethylmaleimide (NEM), followed by removal of the morphine or etorphine by washing. alpha-CAM was still able to bind the pretreated P2 preparations in an irreversible manner. The results indicate that the formation of covalent bonds between alpha-CAM and the SH groups of opioid receptor binding sites is possibly one of the biochemical mechanisms of the irreversible action of alpha-CAM. PMID- 2554673 TI - Regulation of GABA-ergic receptor activity by changes in intracellular calcium: electrophysiological investigation in neurones and endocrine cells. PMID- 2554674 TI - Apparent proton influx via channels: perhaps pHi decrease in depolarised snail neurones is caused by calcium entry. PMID- 2554675 TI - Influence of HCO3- on the postsynaptic actions of GABA at the crayfish neuromuscular synapse. PMID- 2554676 TI - Components of chloride regulation in neocortical neurones. PMID- 2554677 TI - Quinolones and fenbufen interact with GABAA receptor in rat isolated CA1 pyramidal neurons. PMID- 2554678 TI - Calcium accumulation and calcium effects in astrocytes in primary cultures. PMID- 2554679 TI - Characteristics of calcium currents and of pH regulation in Purkinje cells maintained in primary culture. PMID- 2554680 TI - Regulation of secretion in cultured bovine chromaffin cells. PMID- 2554681 TI - Age-related changes in K+ transport and synaptic transmission during and after anoxia in rat hippocampal slices. PMID- 2554682 TI - Osmotic and hormonal stimulation of the third ventricular region of ducks: antidiuretic, circulatory and local neuronal responses. AB - By means of local microperfusion of the 3rd cerebral ventricle, antidiuretic and circulatory responses to stimulations with various hypertonic solutions and norepinephrine were analyzed in conscious ducks. The results suggest ionic rather than osmometric responsiveness of periventricular osmoreceptive elements, which is in line with single unit recordings of periventricular neurons tested in vitro for their osmoresponsiveness. These neurons were located subependymally at the site of greatest responsiveness in vivo, and corresponded to morphologically identified neurons projecting to the neuroendocrine hypothalamo-pituitary system. Antidiuresis was combined with increases in arterial pressure and heart rate in response to hypertonic stimulations with monovalent cations; divalent cations produced long-lasting antidiuresis and equivocal circulatory responses. Norepinephrine elicited antidiuresis which was accompanied by arterial hypotension and bradycardia. Osmotically and norepinephrine induced antidiuresis was combined with increases of plasma ADH concentration. Different modulatory actions of intrinsic adrenergic, angiotensinergic and vasotocinergic neurons are suggested in hypothalamic control of autonomic functions. PMID- 2554683 TI - Bengt Andersson's pioneering demonstration of the hypothalamic "drinking area" and the subsequent osmoreceptor/sodium receptor controversy. PMID- 2554684 TI - Potential use of tuftsin in treatment of candida peritonitis in a murine model. AB - A major complication of continuous ambulatory peritoneal dialysis (CAPD) is peritonitis caused by Candida albicans. Increasing the activity of the peritoneal macrophages, the predominant cell type found in the peritoneal cavity, may be a promising treatment for this infection. Tuftsin was found to increase thioglycollate-elicited mouse peritoneal macrophage activity. 2x10(-7) M tuftsin enhanced two-fold cell association with radiolabelled candida, superoxide aniom production, and killing activity. Thus, a model consisting of mice undergoing peritoneal dialysis was developed in order to study the use of tuftsin as a therapeutic drug against peritoneal candidiasis. Administration of tuftsin (50 micrograms/mouse) before candidiasis induction with a lethal dose of candida (7x10(8) candida per mouse) improved mouse survival up to 70%, compared with 10% in the control group. The potential of tuftsin as a treatment for candidiasis was shown when the infection was induced with a sublethal dose of candida. Daily intraperitoneal injections of tuftsin (50 micrograms) to the sublethally infected mice caused a significant decrease in the number of candida recovered from the peritoneal cavity and from the blood (from 700 +/- 190 to 110 +/- 26 CFU/ml and from 100 +/- 26 CFU/ml to 17 +/- 8 CFU/ml, respectively). In addition, a larger number of peritoneal macrophages with greater phagocytic and killing activity were found in the tuftsin-treated mice. The effect of tuftsin may promote its potential use in the therapy of peritonitis in patients undergoing chronic peritoneal dialysis. PMID- 2554685 TI - Parathyroid hormone-induced ornithine decarboxylase activity in fetal rat osteoblasts. AB - Induction of ornithine decarboxylase (ODC, E.C. 4.1.1.17) activity by parathyroid hormone (PTH) in cultured fetal rat osteoblasts was studied. PTH induced ODC activity and stimulated cAMP production in a dose-dependent manner, the ED50 for cAMP being five times as high as that for ODC. Induction of ODC activity by PTH was partly inhibited by actinomycin D and cycloheximide, with 40 and 55% inhibition, respectively. PTH increased the intracellular ionized calcium concentration ([Ca2+]i), which was absent in a Ca2+-free medium. Blocking calcium influx, lowering the extracellular calcium concentration, and adding trifluoperazine inhibited both induction of ODC activity and stimulation of cAMP production by PTH. A23187 (100 nM and 1 microM), combined with a low dose of PTH (4 nM), resulted in a synergistic induction of ODC activity and an inhibition of cAMP production. A23187 inhibited induction of ODC activity as well as stimulation of cAMP production by the dose of PTH (20 nM) maximally effective in inducing ODC activity. Forskolin together with this maximal dose of PTH resulted in an additive effect on ODC activity and a synergistic stimulation of cAMP production. The current results show similarities and differences with respect to results obtained with osteoblasts from other species and osteoblast cell lines. The present data indicate that (1) PTH stimulates ODC activity and this is partly due to new enzyme synthesis; (2) calcium is involved in induction of ODC activity and stimulation of cAMP production by PTH; furthermore, it is suggestive that calmodulin and/or protein kinase C are involved; and (3) stimulation of cAMP production by PTH depends on an optimal intracellular calcium concentration range. PMID- 2554686 TI - G protein-dependent activation of a phosphoinositide-specific phospholipase C in UMR-106 osteosarcoma cell membranes. AB - Recent evidence suggests that guanyl nucleotide binding (G) proteins are involved in receptor-mediated bone resorption and in osteoblastic function, but the nature of the G protein coupled to effectors that are involved in these skeletal effects is unknown. The purposes of this study were to determine (1) whether a G protein mediates activation of phosphoinositide-specific phospholipase C in UMR-106 rat osteosarcoma cells, and (2) whether parathyroid hormone (PTH) and a PTH-like protein (PLP) associated with humoral hypercalcemia of malignancy promote GTP dependent PIP2 hydrolysis. Addition of GTP (10(-4) M) or guanosine 5'-0-(3 thiotriphosphate, GTP gamma S, 10(-5) M) to membranes prepared from UMR-106 cells labeled with [3H]myo-inositol increased both [3H]inositol trisphosphate (IP3) and [3H]inositol bisphosphate (IP2) formation. The increases in [3H]IP2 and [3H]IP3 produced by GTP were 8.6- and 4.3-fold, respectively. GTP gamma S produced a 17.6 and 11.9-fold increase in [3H]IP2 and [3H]IP3, respectively. The stimulatory effects of GTP and GTP gamma S were dose dependent (GTP ED50 = 3.9 x 10(-6) M; GTP gamma S ED50 = 2.5 x 10(-7) M) and progressive over 10 minutes and required the presence of Mg2+.GTP (10(-4) M) and GTP gamma S (10(-5) M) decreased membrane [3H]phosphoinositides concomitantly with increased [3H]IP2 and [3H]IP3. The GDP analog guanosine 5'-O-(2-thiodiphosphate, GDP beta S) alone did not alter [3H]IP2 or [3H]IP3 production but at 10(-4) M blocks the stimulatory effects of GTP and GTP gamma S. NaF (3 x 10(-2)M) produced a 2.8- and 2.0-fold stimulation of [3H]IP2 and [3H]IP3, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554687 TI - Identification and characterization of parathyroid hormone receptors in rat renal cortical plasma membranes using radioligand binding. AB - Parathyroid hormone (PTH) receptors have been described in renal tissue from several species, but not in the rat. In this study, radioligand binding techniques were used to identify and characterize PTH receptors in rat kidney cortical membranes. The sulfur-free PTH analog [Nle8,18Tyr34]bovine PTH-(1 34)amide was iodinated using the iodogen method. This ligand was suitable for use in identifying PTH receptors in canine renal membranes, but not rat renal membranes. Synthetic, unsubstituted rat PTH-(1-34) was iodinated using the milder, lactoperoxidase technique and was purified by HPLC on a C8 column. [125I]rat PTH-(1-34) bound rapidly to both rat and dog renal membranes. At 22 degrees C reaction reached steady state within 20 minutes, and this level was maintained for at least 3 h. Specific binding was routinely greater than 90% for rat kidney and greater than 95% for dog kidney. Similar results were obtained at 4 degrees C with a longer time required to attain steady state (approximately 45 minutes). Binding was reversible as demonstrated by dissociation of bound ligand after either infinite dilution or displacement with excess nonradioactive PTH. Binding was saturable and of high affinity (rat kidney: Bmax = 2.3 pmol/mg protein, Kd = 3.1 nM, dog kidney: Bmax = 2.1 pmol/mg protein, Kd = 3.7 nM). Rat renal cortical adenylate cyclase activity was stimulated by rat PTH in a dose dependent manner with an EC50 of 4 nM, a value in good agreement with the binding data. This study demonstrates the feasibility of identifying and characterizing parathyroid hormone receptors in rat renal cortical plasma membranes using radioligand binding techniques. PMID- 2554688 TI - Structure-activity relationships of parathyroid hormone analogs in the opossum kidney cell line. AB - Structural alterations in the parathyroid hormone (PTH) molecule produce marked changes in biologic activity. We examined the relative sensitivity of PTH stimulated cAMP formation and PTH-inhibitable Na+-dependent phosphate transport responses to bovine PTH analogs [bPTH-(1-34), bPTH-(1-84), 8,18-norleucine-34 tyrosinamide bPTH-(1-34), bPTH-(7-34)-amide, 8,18-norleucine-34-tyrosinamide bPTH (3-34), transaminated bPTH-(1-34)] and the human PTH-related peptide of malignancy (1-34) in cultured opossum kidney cells. The rank order of potency for stimulation of cAMP formation was bPTH-(1-34) = hPTHrP-(1-34) greater than nle bPTH-(1-34) greater than bPTH-(1-84) much greater than TAbPTH-(1-34). Nle bPTH-(3 34) and bPTH-(7-34) did not affect cAMP formation in intact cells at concentrations up to 10 microM. The rank order of potency for the inhibition of phosphate transport was bPTH-(1-34) = hPTHrP-(1-34) greater than nle bPTH-(1-34) greater than bPTH-(1-84) = TAbPTH-(1-34) greater than nle bPTH-(3-34). TAbPTH-(1 34) was a full agonist and inhibited phosphate transport at concentrations that did not increase cAMP formation, but nle bPTH-(3-34) was a partial agonist in spite of its inability to stimulate cAMP formation. Bovine PTH-(7-34) had no effect on phosphate transport. This study indicates that changes in the PTH molecule produce analogs that apparently discriminate between the cAMP stimulating activity and phosphate transport-inhibiting activities of the native hormone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554689 TI - Modulation of responsiveness of the adenylate cyclase system in avian chondroprogenitor cells by pertussis toxin, PTH, and PGE2. AB - Chondroprogenitor cells, derived from avian tibia epiphyseal growth plate, were cultured in vitro. Incubation of these cells with pertussis toxin augmented their cAMP response to parathyroid hormone (PTH), attenuated the response to forskolin, but did not modify the response to PGE2. Pertussis toxin modulation of the cAMP response was accompanied by ADP ribosylation of two proteins with molecular weights of 39 and 40 kD. Using specific antibodies, the 39 kD protein was identified as the inhibitory guanine nucleotide binding protein (Gi) of the adenylate cyclase system. The other ADP-ribosylated protein has not been identified. Preincubation of the chondroprogenitor cells with PTH or PGE2 resulted in time-dependent heterologous desensitization of the cAMP response to a second challenge of either hormone. The cells did not recover from the densitization for at least 18 h after removal of the hormones. PTH and PGE2 treatment did not affect the cAMP response to forskolin and cholera toxin. The PTH-dependent cAMP production was also not altered by forskolin treatment. PTH homologous desensitization was not affected by pertussis toxin treatment, but the heterologous desensitization due to PGE2 was significantly attenuated. These results suggest that exposure of chondroprogenitor cells to PTH and PGE2 results in heterologous desensitization of the cAMP response. The desensitization is not due to changes in the adenylate cyclase activity. The pertussis toxin-sensitive G proteins are involved in the PTH heterologous rather than homologous desensitization of the cAMP response. PMID- 2554690 TI - Expression of calcitonin receptors during osteoclast differentiation in mouse metatarsals. AB - Metatarsal bones of 15-day-old mouse embryos contain proliferative tartrate resistant acid phosphatase (TRAP) negative (-) osteoclast progenitors that progressively differentiate into multinucleated TRAP positive (+) osteoclasts. Using histochemical and autoradiographic techniques, we have examined the expression of calcitonin receptors during osteoclast differentiation in mouse metatarsals. Fresh mouse metatarsals from embryos aged 14-17 days and metatarsals from 15-day-old embryos cultured for 1, 2, 3, and 6 days were stained for TRAP. Calcitonin binding to osteoclasts and their precursors was studied by incubating metatarsals with [125I]salmon calcitonin (sCT) and quantitating grain counts from autoradiographs of tissue sections. Calcitonin receptors first appear on nonproliferating osteoclast precursors, most often just after or simultaneously with the development of TRAP activity. The effect of sCT on the development of TRAP+ mononuclear preosteoclasts was examined by culturing 15-day-old metatarsals in the continuous presence of 5 mU sCT for periods of up to 3 days and quantitating the number of TRAP+ mononuclear preosteoclasts that develop. Calcitonin did not affect the differentiation of osteoclasts up to the stage of the TRAP+ mononuclear preosteoclast. PMID- 2554691 TI - Role of bone and kidney in parathyroid hormone-related peptide-induced hypercalcemia in rats. AB - A protein responsible for the biochemical syndrome similar to primary hyperparathyroidism associated with certain tumors has been recently characterized and its effects at the level of bone and kidney reported. However, the relative role of tubular reabsorption of calcium (Ca) and bone resorption in the pathogenesis of hypercalcemia induced by this factor is still debated. We investigated the effects of a synthetic amino-terminal fragment of parathyroid hormone-related protein [PTHrP-(1-34)] administered chronically by intraperitoneal osmotic minipumps in thyroparathyroidectomized (TPTX) rats. Clearance studies performed on day 6 of treatment after a 24 h fast revealed an increase in renal tubular reabsorption of Ca and a decrease in renal tubular reabsorption of phosphate (Pi), accompanied by an increase in cAMP excretion. PTHrP-(1-34) (90 pmol/h) stimulated bone resorption as evaluated by an increment in fasting urinary Ca excretion. Although the bone resorption inhibitor aminopropylidene diphosphonate fully corrected urinary Ca excretion and reduced plasma Ca from 3.04 +/- 0.07 to 2.44 +/- 0.21 mM (p less than 0.05), this latter value remained considerably higher than in TPTX control rats (1.54 +/- 0.12 mM, p less than 0.01). In contrast, when the agent WR-2721, which is known to decrease the renal tubular reabsorption of Ca by a PTH-independent mechanism, was given, a further drop in plasma Ca and an increase in urinary Ca excretion were observed. These findings are similar to those found in animals implanted with the hypercalcemic Leydig cell tumor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554692 TI - Intratumoural and intraventricular human lymphoblastoid alpha interferon (HLBI) for treatment of glioblastoma multiforme. AB - A series of ten patients with glioblastoma multiforme were treated with human lymphoblastoid alpha interferon (HLBI) as a single therapy after partial surgical resection (5 cases) or stereotactic biopsy (5 cases). Treatment consisted of intratumoural administration of HLBI (15 x 16(6) IU) every month (8 cases) or in the continuous intraventricular infusion of HLBI (1.8 x 10(6) IU daily) in 15-day cycles (2 cases) until rapid growing of the tumour and important neurological deterioration. The treatments were well tolerated. As judged from data from control groups, the patients demonstrated no improvement in mean survival time and follow-up CT-scan showed rapid progression of the tumour in all cases. PMID- 2554693 TI - The effect of delta-9-tetrahydrocannabinol (delta-9-THC) on the release of nonesterified fatty acids in various brain structures. AB - There was studied the effect of repeated administration of delta-9-THC (10 mg/kg per os for two days) on the level of nonesterified fatty acids (NEFA) in the brain structures of laboratory rats. Palmitic acid, stearic acid, oleic acid and arachidonic acid were estimated with the help of gas chromatography after preliminary separation by TLC. After administration of delta-9-THC, the levels of NEFA decreased in the brain cortex, the brain stem and most in the hypothalamus. The NEFA decrease after delta-9-THC administration is in harmony with the functional inhibition of CNS, provoked by this substance. PMID- 2554694 TI - Microbeads and anchorage-dependent eukaryotic cells: the beginning of a new era in biotechnology. AB - Modern methods for the mass cultivation of anchorage-dependent mammalian cells started with the advent of microcarrier technology. Largely for reasons pertaining to their mode of preparation and ease of cultivation, 150-230 microns microbeads have been overwhelmingly adopted and the technology around them developed. To meet high biomass, macroporous microbeads have been developed. Also, the chemistry of the microsupport has been adapted in order to afford better protection of fragile cells to mechanical wear while simultaneously reorienting their differentiation towards the sought aims (production of cytokines, enzymes etc. ...). Future progress depends upon solutions being brought to problems inherent to this new technology (maintenance of steady state conditions of growth etc. ...) as well as to requirements arising from animal cell culture in general (biosensors, bioreactor's design etc. ...). Besides such technical implementations, biology at large is also expected to benefit from the advent of microcarriers in fields as diverse as the preparation of metaphasic chromosomes in bulk, toxicity testing, organ reconstitution following cell transplantation etc. PMID- 2554695 TI - Signal transduction via the B cell antigen receptor: involvement of a G protein and regulation of signaling. AB - The antigen receptors on B lymphocytes, membrane forms of immunoglobulins, transduce signals regulating B cell growth and differentiation by activating a phosphoinositide-specific phospholipase C. In this report, we describe our recent work aimed at understanding this process in greater detail. We have shown that a GTP-binding component is a necessary cofactor in the stimulation of phospholipase C by mIgM. This component has a number of properties in common with the G protein family of receptor-effector coupling components seen in the adenylate cyclase and other signaling systems. For example, analogues of GTP that cannot be hydrolyzed stimulated mIgM-triggered phosphoinositide breakdown, and an analogue of GDP that cannot be converted to GTP inhibited the reactions. Furthermore, aluminum fluoride, which activates known G proteins, also stimulates phosphoinositide breakdown. The G protein that appears to link mIgM to phospholipase C is not one of the well characterized G proteins involved in the regulation of adenylate cyclase or cGMP phosphodiesterase (GS, Gi, and transducin), as judged by its insensitivity to two bacterial toxins that modify these G proteins, cholera toxin and pertussis toxin. Interestingly, analysis of pertussis toxin sensitivity indicates that there are at least 2 distinct G proteins that couple receptors to phospholipase C. For example, the G protein required for chemotactic peptide receptor signaling in neutrophils is sensitive to pertussis toxin, in contrast to the phosphoinositide signaling G protein in B cells. We have also begun to explore the mechanisms by which mIgM signal transduction can be modulated. Stimulation of protein kinase C with phorbol esters or synthetic DG was found to inhibit mIgM-triggered phosphoinositide breakdown. This regulation probably represents a feedback inhibition that would occur with DG produced by phosphoinositide breakdown. Alternatively, there appear to be other signaling pathways that generate DG33, and they could possibly inhibit phosphoinositide breakdown via protein kinase C. This could be an important locus of regulation during B cell activation. For example, other signals could increase or decrease the potency of this feedback inhibition, and thereby adjust the sensitivity of the B cell to antigen. Alternatively, other agents could stimulate protein kinase C directly, or could stimulate another protein kinase which can do the same thing in this regard, and thereby make the B cell insensitive to antigen by preventing antigen receptor signaling.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2554696 TI - G-protein regulation of polyphosphoinositide breakdown in B cells. PMID- 2554697 TI - Microbial superoxide dismutases. PMID- 2554698 TI - Control of the immune response at the level of antigen-presenting cells: a comparison of the function of dendritic cells and B lymphocytes. PMID- 2554699 TI - The role of viral infections in intrinsic asthma: activation of neutrophil inflammation. PMID- 2554700 TI - [Effects of 5-trifluoromethyl-2'-deoxyuridine (F3TdR) on ultrastructural localization of viral antigens in cultured cells infected with herpes simplex virus (HSV)]. AB - Changes in localization of virus-associated antigens induced by F3TdR were studied using HSV-1 infected monolayer of cultured SIRC cells. A direct enzyme antibody method was employed for preembedding for immunoelectron and light microscopy. At 3 hours post infection (h.p.i.), immunoreaction products began to appear in cytoplasm and nuclei in both the control and the F3TdR-treated group. The amounts increased during the subsequent hours of observation. In the control group, there was an intense diffuse immunoreaction in the cytoplasm and nuclei and dense accumulations of the reaction products were seen on the cell surfaces. In F3TdR-treated cells, intranuclear antigens were present as diffuse accumulations of granular deposits, and cytoplasmic stainings were very much reduced in degree; deposits on the cell surface were a rare finding. Progeny virus particles were significantly fewer in the presence of F3TdR; nucleocapsids were found but in a greatly reduced number and no enveloped particle was found. The morphology of individual nucleocapsid, however, was not significantly altered by treatment with F3TdR. A preponderance of antigenic substances in the nuclei over the cytoplasm in treatment group may reflect distorted protein synthesis and may provide morphological evidence of the biochemically assumed hypothesis of F3TdR efficacy; F3TdR exerts it's antiviral effects through abnormal protein synthesis and resultant defects in capsid assembly. Decreased antigenic substances in the membranous structures of cytoplasm may be related to reduced or abnormal production of viral antigens closely associated with envelopment. PMID- 2554701 TI - [A case of bilateral testicular tumor]. AB - A case of bilateral germ cell tumor of the testis is reported. In 1988, a 36-year old male presented with painless swelling of left scrotal contents. Right orchiectomy and retroperitoneal lymph node dissection for embryonal carcinoma had been performed 5 years earlier. Left orchiectomy was performed and its histological finding was seminoma and embryonal carcinoma. Evaluations including CT scan and Ga scintigraphy revealed no metastasis. Postoperatively, the patient was treated with PVB therapy. Previous reports of bilateral germ cell testicular tumor were reviewed, and the age distribution, interval, and histological classification of these cases are discussed. PMID- 2554702 TI - CT of pineal tumors and intracranial germ-cell tumors. AB - We reviewed 59 cases of pineal tumors and intracranial germ-cell tumors. Most pineal tumors occurred in the first three decades of life, with the exception of pineocytomas, which were seen at a mean age of 34. A male preponderance was noted in the various pineal tumors, except for pineocytomas. The most common tumor of the pineal region was germinoma, which usually showed high density with homogeneously intense enhancement after IV administration of contrast medium. An increased prevalence of pineal calcification was also a feature of pineal germinomas. No characteristic CT features could be found to differentiate among pineal choriocarcinoma, germinoma, embryonal carcinoma, yolk-sac tumor, pineocytoma, and pineoblastoma. CT is useful in detecting intracranial tumors, but the definite diagnosis should depend on pathologic examination and detection of tumor markers in the serum and CSF. PMID- 2554703 TI - Multilocular cystic nephroma: MR findings. PMID- 2554704 TI - Arterial chemoinfusion and embolization of iliac metastasis from pancreatic islet cell carcinoma. PMID- 2554705 TI - Eosinophil and basophil hypersensitivity to deuterium oxide in atopic asthmatic patients. AB - Reaction of basophils and eosinophils obtained from venous blood of 35 atopic asthmatics to deuterium oxide was examined "in vitro". Eosinophils were isolated on sodium metrizoate according to Day. Preparation of basophils by centrifuging and washing in Tyrod's solution was performed according to Benveniste. Eosinophils and basophils were degranulated in D2O at the concentration of 44%. Cells were counted in a Fuch-Rosenthal haemacytometer. Basophil degranulation was observed in almost all (25) of patients in which degranulation of eosinophils had been stated. Marked correlation of reaction to D2O between both types of granulocytes was found. PMID- 2554706 TI - Inhibition of the zymosan-induced chemiluminescence of human phagocytes by adenosine, polyadenylic acid and agents influencing adenosine metabolism. AB - The zymosan-induced chemiluminescence of human phagocytes (neutrophil granulocytes) could be inhibited by adenosine, polyadenylic acid, dipyridamole (an agent blocking the cellular adenosine uptake) and coformycin (a highly potent inhibitor of enzyme adenosine deaminase). It is suggested that in estimating the antianginal and antithrombotic action of dipyridamole, its action on the phagocytes should be taken into account. The same consideration would be true for 2'-deoxycoformycin (a coformycin analogue) used as a potent anticancer and immunosuppressive drug. PMID- 2554707 TI - Lung cancer: detection, prevention, and therapeutics. AB - Lung cancer remains the leading cause of cancer death in the U.S. adult population, and the American Cancer Society estimates that cigarette smoking is responsible for about 83% of all lung cancer cases. It is unlikely that significant reductions in the incidence and mortality associated with lung cancer will be realized without effective antismoking campaigns. Surgery, radiation therapy, and chemotherapy all play a role in the management of lung cancer. At the current time, NSCLC is generally curable only if it is diagnosed while the tumor is small and localized and can be surgically removed. Radiation therapy may also be effective in localized cases. Combination chemotherapy regimens have not consistently produced quality responses in patients with advanced tumors; however, newer regimens utilizing high doses of cisplatin, mitomycin and vinca alkaloids, or cisplatin and etoposide have produced encouraging results. In contrast to NSCLC, SCLC is responsive to combination chemotherapy regimens. Although many different agents are effective in this disease, most small-cell tumors eventually become refractory to chemotherapy and most patients do not survive longer than two years. Although progress has been made in the understanding and management of lung cancer, effective therapies that consistently produce major and durable responses are still lacking. Clinical trials must continue to evaluate therapeutic modalities for all types of lung cancer. PMID- 2554708 TI - Pharmacologic profile of amlodipine. AB - Amlodipine is a potent calcium antagonist, inhibiting Ca2+-induced contractions of depolarized rat aorta with an IC50 of 1.9 nM. Unlike nifedipine, it displayed very slow association and dissociation with the calcium channel. The ability of amlodipine to inhibit Ca2+-induced contractions was strongly dependent on the K+ concentration present before the contraction, suggesting marked voltage dependence of action. Radioligand-binding studies in cardiac membrane preparations suggested that amlodipine may interact directly with both 1,4 dihydropyridine and diltiazem-binding sites on the calcium channel. Hemodynamic studies in anesthetized and conscious dogs showed that amlodipine is a coronary and peripheral vasodilator with a slow onset and long duration of effect, even when given by intravenous injection; the reflex stimulation of cardiac output, heart rate and myocardial contractility induced by amlodipine was attenuated by propranolol, but no marked negative inotropic or dromotropic effects were observed. Amlodipine was an effective oral antihypertensive agent in rat and dog models of hypertension, and its 24-hour duration of action in hypertensive dogs correlated well with its long plasma half-life in this species. The natriuretic properties displayed by amlodipine may contribute to its use as a first-line drug for the treatment of hypertension. PMID- 2554709 TI - Vascular effects of calcium antagonists: implications for hypertension and other risk factors for coronary heart disease. AB - All calcium antagonists (CAs) so far developed are vasodilators, and this property is a most important component of their therapeutic potency in hypertension and angina pectoris. At a cellular level, the specific interaction of CAs with transmembranous calcium fluxes involves both potential and receptor operated channels, respectively. Both alpha 2 and alpha 1 adrenoceptors when activated with an appropriate agonist can trigger the calcium influx through receptor-operated CA channels, alpha 2 adrenoceptors probably more readily so than alpha 1. More recently, angiotensin II receptors have also been demonstrated to be involved, although moderately, in the influx of calcium ions from the extracellular space. The hemodynamic profile of CAs is characterized by a particular specificity for the resistance vessels and for the coronary arterial system, as a useful basis for their therapeutic effect in hypertension and in angina pectoris. The weak natriuretic activity of CAs, probably the result of a tubular effect in the kidney, counteracts the fluid retention to be expected for vasodilator drugs. Interesting ancillary properties of CAs are their potentially favorable effects on the myocardial and vascular hypertrophy associated with long standing hypertension, as well as their antiatherogenic activity that so far has only been demonstrated in animal models. Such additional properties are of potential benefit and deserve further research, since most large-scale hypertension trials have shown that vasodilatation and the reduction of elevated blood pressure as such, are probably not sufficient to adequately protect hypertensive patients against coronary events. PMID- 2554710 TI - Effects of calcium antagonists on lipids and atherosclerosis. AB - The arterial accumulation of cholesterol and calcium is a hallmark of atherosclerosis. Calcium antagonists (CAs) lessen the severity of experimentally induced atherosclerosis in cholesterol-fed animals. The reduction of aortic cholesterol is one of the most striking findings. This effect is achieved without a reduction of plasma lipid or blood pressure, and is probably related to an interference of CAs with lipid metabolism in the arterial wall. To what extent these properties of CAs are due to their ability to block calcium channels still remains to be addressed. This report briefly discusses the available in vivo and in vitro evidence for the antiatherosclerotic properties of CAs, and outlines the possible mechanisms by which these compounds affect cellular lipid metabolism. PMID- 2554711 TI - Calcium antagonists: review and perspective on mechanism of action. PMID- 2554712 TI - Cellular electrophysiology of amlodipine: probing the cardiac L-type calcium channel. AB - The electrophysiologic properties of amlodipine in single guinea-pig ventricular cells were investigated. The degree of ionization of the drug molecule was found to affect both the development of and the recovery from block of L-type calcium channels. Under alkaline conditions, when most of the drug is in a neutral form, the actions of amlodipine resemble previously described neutral dihydropyridine (DHP) compounds. Under these conditions, calcium channel block by amlodipine is reversibly regulated by cell membrane potential, i.e., block is more pronounced at voltages positive to -50 mV and completely relieved at voltages negative to 80 mV. When the drug molecule is ionized, block develops very slowly at positive membrane potentials and is very difficult to relieve on returning the membrane potential to more negative voltages. It is concluded that the degree of ionization of the drug molecule limits access to the DHP receptor and that the drug-bound receptor can be titrated by extracellular hydrogen ions. These results place limitations on the location of the DHP receptor within the cardiac sarcolemmal membrane. PMID- 2554713 TI - Hydrophobic calcium channel ligands: methodical problems and their solution. AB - Niguldipine is a 1,4-dihydropyridine derivative that combines L-type Ca2+ channel blocking effects and alpha 1-adrenolytic activity within a single molecule, exemplifying a novel approach in the treatment of hypertension. As niguldipine is a very hydrophobic compound, it (1) readily adsorbs to surfaces of the plastic ware often used in radioligand binding assays and (2) partitions into the hydrophobic membrane compartments. Both phenomena decrease the actual free drug concentration in radioligand-binding assays and lead to gross underestimation of the affinity of niguldipine (and other hydrophobic ligands) for the 1,4 dihydropyridine binding domain of the L-type Ca2+ channel or for alpha 1A adrenoceptors, respectively. Partitioning of the hydrophobic molecules into the membrane phase leads to a dependence of the Ki value on "total receptor" concentration despite mathematic corrections of the experimentally determined IC50 values. The Ki dependence was mimicked by adding denatured membranes (devoid of high-affinity receptor-binding activity) to native membrane preparations. Loss to pipet tips and tubes was avoided by a special dilution protocol. Partitioning into the hydrophobic membrane compartments needed more elaborate correction procedures. PMID- 2554714 TI - Determination of peak trabecular bone density: interplay of dietary fiber, carbohydrate, and androgens. AB - To elucidate mechanisms linking nutrition and sex hormones to magnitude of peak trabecular bone density, we studied 11 normal women aged 19-21 y. Trabecular bone density was related inversely to dietary fiber (r = -0.69, p = 0.02) and carbohydrate (r = -0.70, p = 0.02) and directly to serum free-and-albumin-bound testosterone (fab T) (r = -0.70, p = 0.02) and total testosterone (total T) (r = 0.66, p = 0.03). Dietary fiber was correlated negatively with fab T (r = -0.74, p = 0.009), total T (r = -0.70, p = 0.02), and androstenedione (Adione) (r = -0.72, p = 0.01). Controlling for the effect of fab T or Adione weakened the relationship between dietary fiber and bone density and the relationship was no longer statistically significant. Conversely, controlling for sex hormones did not abolish the effect of carbohydrate on bone density. The contributions of fab T and carbohydrate to bone density were independent. These results suggest that dietary fiber may depress serum androgens which in turn decrease trabecular bone density. Carbohydrate may also depress bone density but independently of sex steroid hormones. PMID- 2554715 TI - Shift from a mixed to a lactovegetarian diet: influence on acidic lipids in fecal water--a potential risk factor for colon cancer. AB - Although there have recently been reports in the literature indicating that vegetarian-type diets are protective against the development of human colon cancer, this is still far from clear. It was also recently indicated that the concentration of acidic lipids in the aqueous phase of stool constitutes a risk factor for the development of colon cancer. Thus, we examined the effect of a change from a mixed to a lactovegetarian diet on this fecal variable. The dietary change caused a decrease in the total concentration of soluble fecal fatty acids (4310 +/- 3020 to 1080 +/- 1040 mumol/L, p less than 0.05) and deoxycholic acid (125 +/- 42 to 73 +/- 35 mumol/L, p less than 0.05). However, there was no change in either the total bile acid concentration in (164 +/- 54 to 107 +/- 41 mumol/L) or the cellular toxicity of (0.94 +/- 0.55 to 1.60 +/- 0.63 mumol/L, relative survival) the aqueous phase of stool. Thus, the consumption of a lactovegetarian diet may reduce certain risk factors of potential significance in colon carcinogenesis. PMID- 2554716 TI - A long-term metabolic study to assess the nutritional value of and immunological tolerance to two soy-protein concentrates in adult humans. AB - Seventeen healthy young adult men participated in a long-term metabolic study (11 wk) to evaluate the tolerance to and protein nutritional value of two commercially produced soy-protein concentrates. Danpro-S (nine subjects) and Danprotex-H 40 (eight subjects) (Aarhus Oliefabrik A/S, Aarhus, Denmark). Each test protein (0.8 g.kg-1.d-1) served as the sole source of dietary nitrogen. Subjects remained healthy, no problems of clinical or metabolic significance appeared, body weight remained constant, and body composition indices (lean body mass by H2(18)O dilution and creatinine excretion) and basal metabolic rate did not change. N balances fluctuated around body N equilibrium. Immunological studies confirmed the absence of any allergic responses in these subjects. It is concluded that these protein concentrates can be consumed as the sole source of dietary protein for protein nutritional maintenance and with excellent tolerance. PMID- 2554717 TI - Prediction of 'high-risk' cervical papillomavirus infection by biopsy morphology. AB - Certain human papillomavirus (HPV) types (such as type 16) have been linked to high-grade precancers and invasive carcinomas of the cervix. However, the accuracy with which morphologic characteristics will predict the presence and type of HPV infection is controversial. Three pathologists independently classified 102 consecutive cervical biopsies with the use of specific criteria and correlated their findings with the presence of HPV 11, 16, and 18 RNA sequences by in situ hybridization. Based on the presence and distribution of nuclear atypia, abnormal mitotic figures, and koilocytosis, biopsies were classified into borderline condyloma, condyloma, borderline cervical intraepithelial neoplasia (CIN) with koilocytotic atypia (CINK), and CIN. Two or more observers agreed on the diagnosis in 96% of cases. HPV 16-related sequences alone were detected in 0% of borderline condylomata, 17% of flat condylomata, 43% of borderline CINK, 67% of CINK, and 77% of CIN lesions. Other HPVs, including those producing signals with more than one probe, were present in 0, 50, 14, 9, and 0% of these lesions, respectively. The authors data suggest that consistent identification of HPV-related cervical disease requires the presence of specific cytologic changes. In the authors' series, when HPV-related disease is present, CIN is the most common lesion and most (71%) contain HPV 16-related nucleic acids. Thus, a high proportion (88%) of histologic abnormalities associated with HPV-16 could be distinguished as CIN by morphologic characteristics alone, and this distinction could be made by most observers. PMID- 2554719 TI - Comparison of four latex agglutination (LA) and three enzyme-linked immunosorbent assays (ELISA) for the detection of rotavirus in fecal specimens. AB - Eighty-two stool specimens obtained from children with gastrointestinal disease were tested for the presence of antigen to rotavirus by latex agglutination (LA) (Virogen (VR), Meritec (MER), Wellcome (WEL), Slidex Rotatest (SRT), and enzyme linked immunosorbent assays (Rotaclone [TRC], Rotazyme II [RTZ], Pathfinder [PTH]). Confirmatory testing was performed by isolation of rotavirus from stool specimens with the use of a shell vial centrifugation, antigen-detection tissue culture amplification method. The sensitivities and negative predictive values of VR, MER, WEL, SRT, TRC, RTZ, and PTH tests were 85, 89, 95, 91, 98, and 100%, respectively. Each test demonstrated 100% specificity and positive predictive values except the SRT, which attained a specificity of 95%. The WEL LA test may be used as a preliminary rapid screening assay following a stat request. The Kallestad PTH ELISA, however, was determined to be the rotavirus antigen detection kit of choice for routine laboratory diagnostic testing. PMID- 2554718 TI - Detection and typing of human papillomavirus in histologic specimens by in situ hybridization with biotinylated DNA probes. AB - Eighty tissue biopsies from 73 women suspected of having papillomavirus (HPV) infection of the lower genital tract were examined by in situ hybridization with biotinylated DNA probes derived from the complete genomes of four HPV types (6, 11, 16, and 18) and restriction analysis of the extracted DNA on Southern blots. In a subset of 52 samples, the in situ test had a 90.4% sensitivity (47 of 52) in detecting the presence or absence of virus, whereas Southern blot analysis detected HPV with a sensitivity of 98.1% (51 of 52). For 51 samples, in which the viral type was determined by restriction analysis, comparison of the signals separately generated by the four probes after in situ hybridization allowed a correct identification of the infecting HPV type in 86.2% (44 of 51) of cases. PMID- 2554720 TI - Low-dose alternate-day corticotropin therapy in the treatment of childhood seizures. PMID- 2554721 TI - Antiviral drugs in pediatrics. AB - Pediatricians are made familiar with antiviral drugs and are provided with specific recommendations for treatment of viral diseases. The antiviral drugs in clinical use today are discussed in terms of their doses, routes of administration, mechanisms of action, established and potential efficacies, and toxicities. These drugs include acyclovir, amantadine, trisodium phosphonoformate, ganciclovir, ribavirin, rimantadine, vidarabine, and zidovudine. Biologic response modifiers, such as interferons, are mentioned briefly in their historical context. Future trends in antiviral therapy are anticipated with mention of the most promising candidate compounds currently in preclinical trials. PMID- 2554722 TI - Change in plasma acetate levels in diabetic subjects on mixed high fiber diets. AB - When 10 diabetic (five insulin-dependent) subjects with reasonable glycemic control were randomized from their usual (U) diet (about 30 g fiber/day) between two different but isocaloric high fiber diets, each sequentially over 6 wk [HF (55 g fiber/day) and HFS (55 g fiber/day with sucrose taken as 15 g three times daily, with meals)], plasma acetate (mmol/L) increased by 35% on both high fiber diets from 0.21 +/- 0.06 (SD) to 0.28 +/- 0.10 (HF) and 0.30 +/- 0.10 (HFS) (both p less than 0.01). This was higher than the random weekly variation in plasma acetate levels of 11% in six other glycemically stable diabetic patients. When the acetate value was expressed as acetate/glucose, i.e., relative to the fasting glucose level (since fasting levels of acetate and glucose consistently positively correlate), the increase was about 80%. Fasting plasma non-esterified fatty acid (NEFA) levels fell during the high fiber diets and correlated negatively with the fasting acetate level (rs -0.51, p less than 0.005). The fall in NEFA and rise in acetate levels appeared greatest with the sucrose supplemented meals. These results suggest increased exogenous contribution to the plasma acetate level from increased colonic fermentation of dietary fiber and raise the possibility of using such measurements in the assessment of compliance to high fiber diets. The fall in NEFA levels associated with raised plasma acetate levels may contribute to the beneficial effects on glycemic control observed with the high fiber diets. PMID- 2554723 TI - Intrahepatic cholesterol stones associated with peripheral cholangiocellular carcinoma: an autopsy case. AB - A rare autopsy case of intrahepatic cholesterol stones associated with peripheral intrahepatic cholangiocellular carcinoma is presented. The patient, a 60-yr-old man, was diagnosed as having peritoneal dissemination of adenocarcinoma, and died of respiratory failure 4 months later. At autopsy, intrahepatic cholangiocellular carcinoma was found at the left lateral segment of the liver. The cancer was extensively disseminated to the peritoneal cavity. In addition, the liver harbored a cholesterol stone (1.0 cm in diameter) impacted in the right peripheral bile duct, as well as tiny cholesterol stones (0.1-0.2 cm in diameter) scattered in the intrahepatic peripheral bile ducts. A few tiny cholesterol stones were also present in the cancerous bile ducts. Although the causal relationship between the intrahepatic cholesterol stones and cholangiocellular carcinoma remains speculative, clinicians should be aware of this association. PMID- 2554725 TI - Dietary intake and colon cancer: sex- and anatomic site-specific associations. AB - A case-control study was conducted in Utah between July 1979 and June 1983 in which 231 cases of colon cancer identified through the Utah Cancer Registry and 391 controls identified through random digit dialing were interviewed. Odds ratios (OR) were calculated comparing the highest exposure categories with the lowest exposure categories. The highest quintile of body mass index (weight (kg)/height (m)2 for males; weight (kg)/height (m)1.5 for females) was associated with increased risk in both males (OR = 2.1) and females (OR = 2.3). In females, total dietary fat (OR = 1.9) and energy intake (OR = 1.5) were associated with an increased colon cancer risk after adjusting for age, body mass index, and crude fiber. Fiber was protective in females (OR = 0.5) after adjusting for age, body mass index, and energy intake, as was beta-carotene (OR = 0.5) after also adjusting for crude fiber. Adjusted risk estimates in males were 2.0 for total dietary fat, 3.8 for polyunsaturated fat, 2.1 for monounsaturated fat, 2.1 for energy intake, 2.5 for protein, 0.3 for fiber, 0.4 for beta-carotene, and 0.3 for cruciferous vegetables. Risk estimates differed by site of cancer within the colon. In males, protein (OR = 3.8) was a risk factor for cancer of the descending colon, while fats (OR = 2.7-8.8) increased the risk of cancer of the ascending colon. The hypotheses that dietary fat increases colon cancer risk while dietary fiber decreases colon cancer risk and that fat and protein may be independently associated with colon cancer risk are supported. PMID- 2554724 TI - Oral contraceptive use and liver cancer. AB - The risk of liver cancer in relation to use of oral contraceptives was evaluated in a hospital-based case-control study conducted in five US cities from 1977 to 1985. Twelve new cases of liver cancer were identified in women aged 19-54 years; five controls selected from among patients hospitalized for acute conditions unrelated to oral contraceptive use were matched to each case on age (five-year categories), date of interview (three-year categories), and geographic location of the hospital. Among nine cases classified as having hepatocellular carcinoma, eight (89%) had used oral contraceptives; only 16 (36%) of 45 matched controls had used oral contraceptives. Among three other cases (two with cholangiocarcinomas and one with liver cancer of undetermined type), all had used oral contraceptives, compared with four of 15 matched controls. The results confirm the strong positive association between oral contraceptive use and hepatocellular carcinoma observed in earlier studies. Such an association is consistent with evidence that oral contraceptive use is associated with benign hepatic tumors in young women. However, the number of cases of liver cancer in the United States that are attributable to oral contraceptive use is probably small, because liver cancer is extremely rare in the United States. PMID- 2554726 TI - Alveolar cell carcinoma complicating sickle cell anemia: a chance occurrence? AB - Alveolar cell carcinoma, a neoplasm associated with pre-existing inflammatory lung disease, has not been reported previously in patients with sickle cell disease. We present such a patient and suggest that chronic pulmonary scarring in this setting predisposed to development of his malignancy. As survival increases in individuals with sickle cell disease, alveolar cell carcinoma must be considered in these differential diagnosis of otherwise unexplained chronic pulmonary infiltrates. PMID- 2554727 TI - Horner's syndrome and demyelinating peripheral neuropathy caused by high-dose cytosine arabinoside. AB - High-dose cytosine arabinoside may benefit patients with refractory acute leukemia. Peripheral nervous system disturbances caused by cytosine arabinoside have rarely been reported. We describe a patient with acute leukemia who developed Horner's syndrome and a severe demyelinating peripheral neuropathy leading to death after receiving high-dose cytosine arabinoside. Peripheral nerve dysfunction is a potentially serious complication of high-dose cytosine arabinoside. PMID- 2554728 TI - Glomerular atrial natriuretic factor receptors in primary glomerulopathies: studies on human renal biopsies. AB - Human renal biopsies are currently used to provide information about morphologic changes, chronicity of disease, patterns of inflammation, and immunoglobulin deposition. This practice has provided only limited insight into functional aberrations and has failed to provided information necessary for disease classification based on pathophysiology. To expand the potential of the renal biopsy in this regard and to determine whether differences in glomerular atrial natriuretic factor (ANF) binding exist in different forms of primary renal disease, quantitative autoradiography and 125I-human ANF (1-28) were used to determine the location and pharmacological characteristics of ANF binding sites in the normal human kidney. Specific ANF binding was highest in the glomeruli, but lower levels of specific binding were localized to the inner medulla and the interlobular arteries. ANF binding sites in the human kidney were found to be highly stable and similar in both location and pharmacology to those observed in experimental animals. As determined by saturation experiments, the equilibrium dissociation constants for glomeruli, inner medulla, and interlobular arteries were almost identical at 4.0 x 10(-11) mol/L. Competitive binding inhibition studies with unlabeled human ANF (1-28) demonstrated highly specific binding shared by the glomerulus, inner medulla, and interlobular artery, with apparent half-maximal inhibition concentrations of 9.2 x 10(-10) mol/L, 8.0 x -10 mol/L, and 8.2 x 10(-10) mol/L, respectively. Quantitation of specific binding of ANF to glomeruli in needle biopsy specimens of three primary glomerulopathies, ie, minimal-change disease, membranous nephropathy, and focal glomerulosclerosis, showed no differences among the groups. This study demonstrates the feasibility of studying receptor physiology on biopsy specimens of the human kidney and should allow renal diseases, particularly of glomerular origin, to be characterized according to differences in hormone binding and hormone responsiveness. The absence of significant differences in glomerular ANF binding in the primary glomerulopathies studied is consistent with other studies that have failed to delineate important pathophysiological differences in renal function and volume homeostasis in these disease states. PMID- 2554729 TI - CAMFAK syndrome: a demyelinating inherited disease similar to Cockayne syndrome. AB - CAMFAK syndrome is an inherited disease characterized by congenital cataracts, microcephaly, failure to thrive, and kyphoscoliosis with onset in early infancy. Its pathogenesis has not been clearly defined. We report on a patient with this syndrome and present evidence that it is a neurologic disease characterized by peripheral and central demyelination similar to that seen in Cockayne syndrome. PMID- 2554730 TI - Effects of uremia, acetate and bicarbonate dialysis on beta-adrenergic responsiveness as assessed by epinephrine-stimulated adenylate cyclase. AB - This study was designed to evaluate beta-adrenergic responsiveness as assessed by the generation of cyclic AMP after the subcutaneous administration of epinephrine in 31 subjects: normal controls, patients with chronic renal insufficiency, hemodynamically stable patients on chronic acetate dialysis and hemodynamically unstable patients with acetate intolerance on chronic bicarbonate dialysis. As compared with controls, only unstable patients on bicarbonate dialysis presented impaired beta-responsiveness, which, in turn, was acutely reduced only after acetate but not bicarbonate dialysis. Our results show that acetate dialysis impairs the beta-adrenergic responsiveness and that the observed beta hyporesponsiveness in unstable patients with acetate intolerance may represent part of a broader spectrum of autonomic dysfunction which may develop in some patients on chronic hemodialysis. PMID- 2554731 TI - Foscarnet nephrotoxicity: mechanism, incidence and prevention. AB - Foscarnet is a pyrophosphate analogue that has been successfully used in severe cytomegalovirus (CMV) infections. Little is known of the incidence and mechanisms of foscarnet-induced nephrotoxicity as most data comes from recipients of renal allografts or from patients with severe underlying disease or with other nephrotoxic drugs. We have retrospectively analyzed the evolution of renal function after 56 courses of foscarnet. In addition, we have prospectively studied the protective effects of hydration on foscarnet nephrotoxicity (2.5 liters of saline/day during the night before the foscarnet therapy and throughout the course of treatment). Foscarnet-induced acute renal failure was defined as a rise in serum creatinine of at least 25% from the basal value. An increase in serum creatinine occurred in 37 cases out of the 56 courses of foscarnet (66%). The mean serum creatinine prior to foscarnet was 80.5 +/- 3.3 mumol/l and the mean increase was 190 +/- 28.3 mumol/l (range 80-1,000). Peak serum creatinine was higher than 200 and 300 mumol/l in 16 and 13 patients, respectively. Kidney obtained at autopsy from a 30-year-old male with AIDS, CMV pneumonitis and acute renal failure secondary to foscarnet administration showed an extensive tubular necrosis. In the group which was prospectively hydrated only 1 patient had an acute renal failure. The mean serum creatinine at the peak (96 +/- 4 mumol/l) and at the end of the treatment (83 +/- 4 mumol/l) was significantly lower (p less than 0.05) than in non hydrated patients. In conclusion, foscarnet is a highly nephrotoxic drug which induces acute tubular necrosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554732 TI - Urine electrolytes in the assessment of extracellular fluid volume contraction. AB - The purpose of this study was to determine which urine electrolytes should be measured to confirm that the extracellular fluid (ECF) volume is depleted. ECF volume contraction was induced by furosemide administration to rats consuming an electrolyte-free diet. An external potassium balance was achieved by replacing potassium losses with KHCO3 and KCl so that the sodium and chloride deficits were comparable (equivalent to a 30% reduction in ECF volume). As expected, the urine sodium and chloride concentrations fell to 2 +/- 0.3 mmol/l and 3 +/- 0.3 mmol/l, respectively. Rats were then randomized to receive 50-75% of their sodium or chloride deficit as either: NaCl (control group), NH4Cl or NaHCO3 to mimic clinical situations associated with ECF volume contraction. In the NaCl group, the urine sodium and chloride concentrations remained low (6 +/- 2 mmol/l and 7 +/- 2 mmol/l), consistent with persistent ECF volume contraction. Although the NH4Cl group continued to have a low urine sodium concentration (2 +/- 0.2 mmol/l), there was now a marked increase in the urine chloride concentration (51 +/- 7 mmol/l; p less than 0.01 vs. NaCl group). In contrast, although the NaHCO3 group continued to have a low urine chloride concentration (2 +/- 1 mmol/l), there was a significant increase in the urine sodium concentration (19 +/- 3 mmol/l; p less than 0.01 vs. NaCl group). We conclude that the clinical assessment of ECF volume by urine electrolytes requires an evaluation of both the urine sodium and chloride concentrations. PMID- 2554734 TI - The perversion of research and the paralysis of action. PMID- 2554733 TI - An electroretinographic and molecular genetic study of X-linked cone degeneration. AB - We evaluated full-field electroretinograms from members of a family with X-linked cone degeneration. The 15-year-old propositus had near normal visual acuity and a protan deficiency. His maternal grandfather and great uncle had a visual acuity of 20/200, a deficiency in color vision, and signs of macular degeneration; all had normal rod electroretinographic responses and diminished cone electroretinographic responses. The mother and maternal aunt of the propositus had normal visual acuity and diminished cone electroretinograms with predominant loss of red cone function. Their cone responses were greater than those of affected males. Genomic DNA isolated from these patients was analyzed with a red cone pigment gene cDNA probe that disclosed a 6.5-kilobase deletion in the red cone pigment gene. These findings substantiate that a defect in a gene encoding for a cone photoreceptor protein can lead to a cone photoreceptor degeneration. PMID- 2554735 TI - Pap smear screening at the workplace. PMID- 2554736 TI - The selection and definition of targeted work-related conditions for surveillance under SENSOR. AB - Lists of reportable conditions and case definitions are important tools for epidemiologic surveillance. As part of an initiative to encourage occupational disease surveillance systems linked to intervention at the state level, we have proposed a list of target conditions and are developing a set of standard epidemiologic case definitions. Experience gained from state health department pilot projects using SENSOR and other surveillance approaches will be used to promote effective condition-specific surveillance strategies on a wider scale. PMID- 2554737 TI - Dietary fiber and giardiasis: dietary fiber reduces rate of intestinal infection by Giardia lamblia in the gerbil. AB - Gerbils were maintained on a low-fiber (5%) or a high-fiber (20%) diet in which the major fiber source was cellulose. Animals in the low-fiber diet group were significantly more likely to become infected when inoculated with 100 Giardia lamblia cysts than were animals in the high-fiber group. No differences were detected in gastrointestinal transit, gastric, and small intestinal luminal pH, or in duodenal mucus blanket acidic glycoprotein between animals in the high- and the low-fiber diet groups at the time of cyst inoculation. The fiber content of the diet after cyst inoculation determined the infection rate. These data suggest that the dietary fiber effect occurred during trophozoite colonization of the small intestine. When infected animals on the low-fiber diet were placed on the high-fiber diet for 24 hr, trophozoite clearing occurred in the lower small intestine. In the jejunum, the number of trophozoites attached to the mucosal surface decreased, while the number associated with luminal mucus increased. We conclude that the fiber-induced mucus secretion and the bulk movement of the insoluble fiber reduced the attachment of trophozoites to the intestinal mucosa, which decreased the probability of trophozoites establishing and sustaining colonization of the mucosa. PMID- 2554738 TI - An "erg" of epidemiology. PMID- 2554739 TI - [Endogenous beta-adrenomimetic substance as a constituent of the adrenergic beta receptor blocking mechanism]. AB - In order to reveal endogenous beta-adrenomimetic in the blood the authors proposed the biological testing of the serum and plasma on the isolated myometrium of the rat. For this purpose obsidan was used as a beta-adrenoblocker. It was shown that endogenous beta-adrenomimetic different from adrenaline was present in the blood of males, nonpregnant and pregnant females and parturients. Besides, its concentration increased in pregnancy and remained high early in labor. PMID- 2554740 TI - ACTH-induced seizures in an infant with West syndrome. AB - We report seizures induced by adrenocorticotropic hormone (ACTH), which were demonstrated clinically and electro-encephalographically, in a severely handicapped 7-month-old infant with West syndrome due to perinatal hypoxicischemic encephalopathy. Although tonic spasms (original seizures) decreased soon after starting ACTH treatment, new brief tonic seizures, somewhat more slowly motioned than the original tonic spasms, frequently appeared only during sleep after consecutive ACTH injections for 11 days, in place of the tonic spasms seen in the waking state. After discontinuation of ACTH therapy with the last injection on the 16th day, the brief tonic seizures began to decrease and finally disappeared in 8 days. Ictal EEG of new brief tonic seizures revealed diffuse fast spiky wave bursts, 50-150 microV and 10-20 c/s, with a duration of 0.5-4 seconds, which were different from attenuation associated with low voltage rhythmic fast activity corresponding to tonic spasms, the original seizures. Therefore, we considered that the new brief tonic seizures, which appeared only during sleep in the course of ACTH therapy, were ACTH-induced seizures. PMID- 2554741 TI - Intraepidermal morphologic manifestations in lysosomal diseases. AB - This paper reports the ultrastructural findings for the epidermis of biopsied skin specimens in numerous lysosomal diseases, which can be grouped as follows: a) presence of vacuolar lysosomal residual bodies in mucopolysaccharidoses I, II and III, Salla disease, GM1-gangliosidoses and infantile type II glycogenosis; b) avacuolar lysosomal residual bodies in Niemann-Pick disease type C, mucolipidosis IV, Farber disease, Fabry disease, and late infantile and juvenile neuronal ceroid-lipofuscinoses; c) absence of lysosomal residual bodies in GM2 gangliosidoses, metachromatic leukodystrophy, Gaucher disease and sialidosis type III. Whenever possible, a biopsy of the skin for morphological diagnosis of lysosomal disorders ought not to be confined to the epidermis. PMID- 2554742 TI - High-dose intravenous methyl-prednisolone in chronic relapsing dysimmune polyneuropathy (CRDP) PMID- 2554743 TI - Management of atopic dermatitis. PMID- 2554744 TI - Infections as contributing factors to atopic dermatitis. AB - Certain cutaneous viral infections, such as Herpes simplex, vaccinia and varicella infections are known to occasionally run an usually severe course in AD. Patients with AD display increased frequencies of recurrent cold sores and Herpes zoster and they also have increased antibody titers to Epstein-Barr virus. Heavy colonization of the skin with staphylococci and streptococci is common. The findings of increased severity and/or frequencies of these infections in AD may be explained by dysfunctional cell-mediated immunity and by cutaneous changes associated with AD. There is suggestive, but not any firm evidence, that infections play a causal role in the precipitation and exacerbation of AD. Infections are thus, in most cases, probably consequences rather than causes of the disease. PMID- 2554745 TI - [Diagnostic role of the biopsy in angiofibroma of the cavum]. AB - The AA. inform about the case of a child, 13, with an antro-choanal Polyp of angiofibromatous structure, diagnosed as Angiofibroma. Though in the greater number of instances the arteriography is enough to support the suspicion, the confirmation through histopathology is compulsory. PMID- 2554746 TI - [Tumors of the jugular and tympanic glomus. Unusual locations. Magnetic resonance imaging in the diagnosis. Comments on 3 cases]. AB - Inform about the last contributions to the subject published. The importance of the N.M.R. and its place respect to the T.A.C. is considered. Report of cases localized and behaving atypically lately published are reviewed. Other 3 cases diversely localized in the glomus system tympano-jugularis, as well as its diagnosis, treatment and evolution are included. Only those cases spreading to the endocranium are excluded. PMID- 2554747 TI - Immobilization of proteins on oxidized crosslinked Sepharose preparations by reductive amination. AB - Mild periodate oxidation of certain commercially available crosslinked agarose beads (Sepharose CL-4B and CL-6B) results in the generation of aldehydo groups which were useful for immobilization of amino compounds by reductive amination using pyridine borane. Consumption of periodate ion and production of formaldehyde were only observed with crosslinked Sepharose preparations and were correlated with a binding capacity much greater than that of uncross-linked gels when subjected to the reductive amination reaction. Up to 50 mg (approximately 0.73 mumol) of bovine serum albumin and 30 mumol of glycylglycine were coupled per gram of moist oxidized Sepharose CL-6B. The immobilization reaction was shown to proceed at neutral pH requiring about 12 h for completion and to be relatively insensitive to temperature and pyridine borane concentration. The oxidized gel was shown to be stable for at least 2 months upon storage in 0.1 M acetic acid. This method has proven to be useful for the preparation of a variety of affinity matrices and immobilized enzymes. PMID- 2554748 TI - A thiophilic adsorbent for the one-step high-performance liquid chromatography purification of monoclonal antibodies. AB - Thiophilic adsorption chromatography, developed originally by Porath and colleagues (1985, FEBS Lett. 185, 306-310) for conventional chromatographic techniques, was transformed to the HPLC mode upon preparation of new and improved sulfur-containing silica beads. The new thiophilic adsorbent is of high capacity and is suitable for the rapid and single-step purification of all subclasses of monoclonal and polyclonal antibodies. Due to its broad specificity, the thiophilic silica column is an efficient, stable, and inexpensive substitute for protein A and protein G columns used today to purify antibodies. PMID- 2554749 TI - Immunoprecipitation of labeled antigens with Eupergit C1Z. AB - Eupergit C1Z, a nonporous sedimentable acrylic polymer is shown to be a convenient solid support for isolation of antigens with high purity from small amounts of detergent-solubilized cells. Several applications of Eupergit C1Z coupled with antibodies and anti-antibodies are described, and compared with two other methods of immunoprecipitation. PMID- 2554750 TI - A spectrophotometric procedure for measuring oxoglutarate and determining aminotransferase activities using nicotinamide adenine dinucleotide phosphate linked glutamate dehydrogenase from algae. AB - A new spectrophotometric procedure is described for determining glutamate dependent activities of aspartate aminotransferase, alanine aminotransferase, and ornithine aminotransferase with NADPH-linked glutamate dehydrogenase (GDH) from nitrate-grown Stichococcus bacillaris. The algal NADPH-GDH is highly specific for oxoglutarate and can catalyze the reduction of this keto acid in the presence of high glutamate concentrations, and thus is suitable for the measurement of oxoglutarate produced in glutamate-dependent amino-transferase reactions. The alga produces large amounts of NADPH-GDH which can be adequately purified in a few simple steps. The purified enzyme can be stored at 4 degrees C for several weeks without any detectable loss of activity. The algal NADPH-GDH can also be used for the estimation of small amounts of oxoglutarate in aqueous extracts. PMID- 2554751 TI - Enzymatic assay for deoxyribonucleoside triphosphates using synthetic oligonucleotides as template primers. AB - The enzymatic assay for deoxyribonucleoside triphosphates has been improved by using synthetic oligonucleotides of a carefully defined sequence as template primers for DNA polymerase. High backgrounds, which limit the sensitivity of the assay when calf thymus DNA or alternating copolymers are used as template primers, were eliminated with these oligonucleotide template primers. Sensitivity was further increased by designing the template primer to incorporate multiple labeled deoxyribonucleotides per limiting unlabeled deoxyribonucleotide. Each of several DNA polymerases exhibited unique reaction characteristics with the oligonucleotide template primers, which was attributed to the differing exonuclease activities associated with these various enzymes. Assay optimization therefore included matching the polymerase with the template primer to obtain the lowest background reaction and highest sensitivity. This modified assay is particularly well suited for keeping cell sample size to a minimum in experimental protocols which generate large numbers of data points or require careful timing of sampling. With this technique, we measured the levels of all four deoxyribonucleoside triphosphates in extracts from as few as 2 x 10(4) cultured cells. PMID- 2554753 TI - Use of peptide substrates for affinity purification of protein-serine kinases. AB - The ability of protein kinases to phosphorylate synthetic peptides corresponding to identified protein phosphorylation sites has previously been used to determine primary structural requirements and has helped define distinct "recognition sequences" for a variety of enzymes. Here, we have used an immobilized synthetic peptide derived from glycogen synthase to specifically purify two protein kinases. In the case of one, glycogen synthase kinase-3, the peptide is only a substrate if previously phosphorylated at a distinct site by another protein kinase, casein kinase-II. This prerequisite is reflected in the differential affinity of glycogen synthase kinase-3 for the immobilized phospho- and dephosphopeptide. This difference in binding has been exploited to effect purification of glycogen synthase kinase-3 as well as casein kinase-II. The general applicability of peptide-based affinity chromatography is discussed. PMID- 2554752 TI - Reduction and fluorescent labeling of cyst(e)ine-containing proteins for subsequent structural analyses. AB - Procedures which allow rapid, quantitative, and selective fluorescent labeling of protein cyst(e)ine residues prior to electrophoresis by reaction with 4 (aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (ABD-F) under mild conditions are described. After labeling, the protein(s) of interest is easily monitored throughout electrophoresis and subsequent electroblotting or electroelution procedures. The stoichiometry of labeling and therefore the number of cysteine and/or half-cystine residues can be measured spectrophotometrically or fluorometrically and the derived cyst(e)ine adduct can also be quantitated by amino acid analysis and identified in protein sequencing. N-terminal blockage is not observed under the conditions utilized, nor are any other amino acid side chains modified. The procedures described allow complete, rapid, and facile reduction and alkylation of proteins with simultaneous incorporation of a fluorophore, permitting sensitive detection in subsequent manipulation of the proteins. Quantitative fluorescence prelabeling also allows the generation, purification, and sequencing of peptide fragments containing cyst(e)ine residues for determination of internal sequences and residues involved in disulfide bonds. PMID- 2554754 TI - Purification of human genomic DNA from whole blood using sodium perchlorate in place of phenol. AB - We have developed a new, rapid method for the extraction of human genomic DNA from whole blood samples. Traditionally, genomic DNA has been extracted from blood by overnight proteinase K digestion of lysed peripheral lymphocytes followed by phenol/chloroform extraction. In addition to being time consuming, the use of phenol involves inherent risks due to the toxic nature of the reagent. Our method for the extraction of DNA from whole blood uses sodium perchlorate and chloroform instead of phenol with a significant time savings realized as well as fewer hazards to the technician. Furthermore, DNA prepared by this new method is an excellent substrate for restriction endonuclease digestion and Southern hybridization analysis. PMID- 2554755 TI - Directional immobilization of sodium- and potassium-activated ATPase to expose its cytoplasmic part to the liquid phase on microtiter plates by wheat germ agglutinin. AB - A convenient method for highly efficient and directional immobilization of intact sodium- and potassium-activated ATPase (Na,K-ATPase) using wheat germ agglutinin linked on microtiter plates was developed. Wheat germ agglutinin, which bound tightly to the beta-subunit of Na,K-ATPase and had no effect on the Na,K-ATPase activity, the potassium-activated p-nitrophenylphosphatase activity, or the inhibitory action of ouabain, was covalently linked to microtiter plates and used as an immobilizer of the enzyme. The amount of Na,K-ATPase coupled to microtiter plates in this immobilizing system was more than 10-fold greater than that used in the direct immobilizing system (O. Urayama, M. Nakao, H. Nagamune, and H. Sugiyama, (1984) Anal. Biochem. 141, 194-198). Also in this system, the cytoplasmic domain of Na,K-ATPase was exposed to the liquid phase. This technique was useful for investigating the reactivities of monoclonal antibody specific for the cytoplasmic domain of the enzyme. Moreover, because this technique was used successfully in the immobilization of periodic acid--Schiff positive staining glycoprotein 1 prepared from human erythrocytes and human alpha 2-macroglobulin, the technique should also be useful for other membrane or secreted proteins that possess N-linked sugar chains containing bisecting N-acetylglucosamine or a high amount of sialic acid. PMID- 2554756 TI - Assembly of genes from partially overlapping fragments using single-stranded DNA and sequence-specific synthetic oligodeoxynucleotides. AB - A simple procedure for the precise assembly of functional DNA sequences from overlapping fragments is described. The fragments to be joined are cloned in tandem in the proper relative orientation into a vector from which single stranded DNA copies can be obtained. Single-stranded DNA is cut by a restriction enzyme at corresponding sites in the two overlap regions, which are made double stranded by annealing an oligonucleotide of appropriate sequence to them. This results in the excision of the unwanted sequences between the two overlap regions. After removal of the restriction enzyme the DNA is reannealed using the same oligonucleotide, ligated to give closed circular molecules and used to transform competent cells. Clones with the desired structure appear in the progeny at high frequency. The method has the advantage that restriction enzymes with short recognition sequences, cutting frequently in the target DNA, can be used and hence the overlap region required can be quite short. PMID- 2554757 TI - Rapid isolation of high-molecular-weight DNA from agarose gels. AB - We have developed a simple, reliable, and rapid method for recovering DNA from agarose gels. While many methods for DNA extraction have already been described, few provide quantitative recovery of large DNA molecules. These procedures generally require costly apparatus, extended elution times, or considerable handling of the sample after elution. Our method employs a novel electroelution chamber constructed from acrylic plastic. Gel slices containing DNA are placed in the chamber between platinum electrodes. Voltage is applied and a continuous flow of buffer sweeps the eluted DNA from the chamber into an external receptacle. Elution is complete in 7 min. Concentrated DNA is obtained by butanol extraction and alcohol precipitation in 1 h. Recoveries, quantitated by counting radiolabeled DNA or by densitometry of analytical gels, were 94 to 100% for fragments of 4 to 50 kb. The eluted DNA was undegraded and could be digested with restriction enzymes, ligated, end-labeled, or used to transform cells as efficiently as noneluted DNA. Complete elution of a 100-kb plasmid, a 194-kb concatemer of bacteriophage lambda, and of 440- and 550- chromosomes of Saccharomyces cerevisiae was also achieved using the same process. This method is suitable for routine use in a wide range of cloning applications, including the electrophoretic isolation of large DNA molecules. PMID- 2554758 TI - Quantitation of testicular and somatic cytochromes c in testis and somatic tissues from developing rats. AB - By combining chromatographic and spectral procedures, simple and quantitative assays for somatic cytochrome c (cyt cs) and testicular cytochrome c (cyt ct) in crude animal tissue extracts were developed. Using this assay procedure, limited developmental studies of cyt ct and cs were performed with tissue extracts of 27 , 58-, and 85-day-old rats. Specific contents of cyt cs in somatic tissues (i.e., micrograms of cyt c/g of tissue) of these three age groups did not show significant variations. However, the amounts of both cyt ct and cs in testis were markedly increased as the rats approached maturity. Increasing cyt ct/cyt cs ratios as the rat developed to maturity suggest that expression of cyt ct is preferentially required for specific function of testis. Application of both molecular biological techniques and this assay (for holo-cyt ct) should be useful to study the overall regulation of the expression of cyt ct in testis. PMID- 2554759 TI - A solid-phase extraction procedure for DNA purification. AB - The preparation and use of particulate materials for the removal of proteins from nucleic acid samples by solid-phase extraction procedures are described. The solid-phase extraction procedure is analogous to the classical phenol extraction for DNA purification, with the exception that the phenol is replaced with insoluble particulate materials that are chemically similar to phenol and thus function in an analogous manner. These particulate materials have a very high affinity for proteins and a very low affinity for nucleic acids. With these materials, it is possible to remove large quantities of proteins (i.e., tens of milligrams) from minute quantities (submicrogram) of nucleic acid and quantitatively recover the latter in a biologically active state. Compared to other procedures that are currently used to purify nucleic acids, the protocols using these materials offer the advantages of speed, quantitative DNA recovery, safety, and convenience. PMID- 2554760 TI - Quantitative fluorescence of DNA-intercalated ethidium bromide on agarose gels. AB - Techniques for analyzing DNA distributions on agarose gels are examined by both two-dimensional and one-dimensional methods. It is demonstrated that very large errors in DNA concentration occur in such analyses unless (i) the electrophoresis is performed in a careful, reproducible manner, (ii) the films are calibrated with an internal standard, (iii) high resolution densitometry is used for analyzing the films, and (iv) appropriate background controls are used to determine the baselines for integration. Two-dimensional scanning produces more accurate results than one-dimensional scanning, but in cases where the bands are relatively uniform, the one-dimensional analysis gives good results. A technique for determining accurate distributions is described. PMID- 2554761 TI - A spectroelectrochemical cell designed for low temperature electron paramagnetic resonance titration of oxygen-sensitive proteins. AB - In this paper we describe an anaerobic titrator made virtually from glass with a small amount of high vacuum epoxy mounted directly to a quartz EPR tube. A complete titration may be carried out with as little as 600 microliters of sample. This cell features the anaerobic manipulation of an electrochemically poised solution from an electrochemical pouch to an EPR tube. The cell uses a gold foil working electrode and Ag/AgCl reference and counter electrodes. The reference and counter electrodes are isolated from the sample by leached Vycor glass. In the work reported here, we used this cell to determine the equilibrium redox potential of methyl viologen in an EPR titration. With methyl viologen as an indicator we found that the cell has a residual oxygen level of 1.5 microM with a leak rate of 0.005 nmol/min. After moving the solution into the EPR tube, freezing, performing EPR, and thawing, the potential of the methyl viologen solution drifted only 2 mV. During the titration, the poised potentials were stable, drifting only 1 mV/min. Formal potentials as low as -630 mV in a vitamin B12-type protein have been determined with this cell (S. R. Harder, W.-P. Lu, B. A. Feinberg, and S. W. Ragsdale (1989) Biochemistry, in press). PMID- 2554763 TI - [Effect of the nondepolarizing muscle relaxant atracurium in patients with vascular diseases]. AB - Electric myographic examination was performed in 37 patients with atherosclerotic occlusive lesion in the great vessels. The patients were found to have concurrent impaired neuromuscular transmission that, however, produced no effects on the myoparalytic activity of the nondepolarizing relaxant atracurium (UK). The application of atracurium, 0.5 mg/kg following induction in the anesthesia provided good conditions for tracheal intubation and potentialities of monomyorelaxation during the entire operation. Atracurium given in a starting dose of 0.5 mg/kg and a myoplegia-maintaining dose of 0.25 mg/kg ensured adequate myorelaxation within 30-40 minutes. The lack of cumulative effects made the clinical effect of atracurium readily controlled and required no pharmacological antagonization at the end of surgery. Safe application of atracurium is useful in cardiovascular surgery. PMID- 2554762 TI - Correlation between the sperm motility and the adenylate cyclase activity in infertile men. AB - The relationship between the sperm motility and the adenylate cyclase activity in spermatozoa from patients with male infertility was investigated. Cyclic AMP contents were measured from 34 semen specimens and adenylate cyclase activity were assayed from 50 specimens. All semen specimens were collected from infertile men at our clinic by masturbation after 5 days sexual abstinence. Spermatozoa were washed and concentrated according to the method described by Harrison. The cAMP contents and the adenylate cyclase activity of spermatozoa were determined by radioimmunoassay. Positive correlations were found among the cAMP contents, the adenylate cyclase activity in human spermatozoa and the sperm motility. This result suggests that the sperm motility is regulated by the adenylate cyclase activity via cAMP and that poor sperm motility observed in infertile men is partially caused by the impairment of adenylate cyclase system. PMID- 2554764 TI - [Biology of the Epstein-Barr virus]. AB - Epstein-Barr virus (EBV) displays several biological properties which distinguish it from any other human herpesvirus. The most important of these is that it infects and immortalizes human B lymphocytes, both in vitro and in vivo. These indefinitely growing cells consistently express viral proteins in the nucleus or in the plasma membrane; at least one of these is required for immortalization. In severely immunocompromised hosts such as transplant recipients and AIDS patients, EBV induces B-cell lymphoma. Two strains of EBV (A and B) have recently been recognized; they behave differently in their reaction to EBV-specific cytotoxic T lymphocytes. Further studies are needed to define more accurately the part played by EBV in Burkitt's lymphoma and nasopharyngeal carcinoma. Besides immortalization, EBV can infect oropharyngeal epithelial cells in vivo, replicating in them at the time of primary infection (infectious mononucleosis) and reactivations. In fact, EBV infects most people throughout the world while rarely causing severe disease. PMID- 2554765 TI - Ventilation-perfusion mismatching in chronic obstructive pulmonary disease during ventilator weaning. AB - Using the multiple inert gas elimination technique, we studied ventilation perfusion (VA/Q) relationships in eight patients with chronic obstructive pulmonary disease (COPD) during mechanical ventilation (MV) and again during weaning (spontaneous ventilation [SV] through an endotracheal tube) from MV needed for acute respiratory failure. The patients, seven men and one woman with a mean age of 63 +/- 2.8 (SEM) yr (FEV1 33 +/- 5.2% of predicted), required MV for 9.0 +/- 2.4 days prior to the study. The patients were studied at maintenance FIO2 (0.28 to 0.40) while breathing 100% O2, both during MV and SV. After 30 min of SV, PaCO2 increased from 48.9 +/- 3.4 to 58.3 +/- 3.1 mm Hg (p = 0.003) and pH decreased from 7.42 +/- 0.01 to 7.36 +/- 0.01 (p = 0.001) without significant changes in PaO2. Despite a decrease in tidal volume (VT) from 700.0 +/- 41.1 during MV to 313.0 +/- 39.6 ml during SV (p = 0.001), minute ventilation remained unchanged (from 8.2 +/- 0.7 during MV to 7.4 +/- 0.6 L/min during SV). Furthermore, cardiac output (QT), oxygen delivery (QO2), and mixed venous PO2 (PVO2) significantly rose during SV when compared with the MV (QT: from 4.7 +/- 0.4 to 6.7 +/- 0.7 L/min, p = 0.011; QO2: from 857.3 +/- 113.0 to 1078.5 +/- 158.9 ml/min, p = 0.0074; PVO2: from 36.7 +/- 1.1 to 42.3 +/- 2.2 mm Hg, p = 0.041). Overall VA/Q inequality worsened as blood flow was redistributed to low VA/Q areas (from 9.4 +/- 4.4 to 19.6 +/- 5.3% of QT, p = 0.05). The dispersion of the ventilation distribution (log SDV) significantly worsened during SV (from 1.0 +/- 0.08 during MV to 1.2 +/- 0.08 during SV, p = 0.044). No changes were observed in either series dead space or ventilation of high VA/Q ratio units.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554766 TI - A specific LTD4/LTE4-receptor antagonist improves pulmonary function in patients with mild, chronic asthma. AB - LY171883 is a new selective LTD4/LTE4-receptor antagonist. To evaluate the efficacy of LY171883, we studied 138 nonsmoking asthmatic patients, 18 to 65 yr old, in a double-blind, randomized block-design study. All patients were required to demonstrate a greater than or equal to 15% increase in FEV1 after inhaled bronchodilator use and were then randomly assigned to either LY171883 (600 mg) or placebo twice daily for 6 weeks. Assessment of efficacy was measured by inhaled metaproterenol use (mg/wk), symptoms, twice-daily peak expiratory flow, and weekly FEV1 measurements. LY171883-treated patients had improved FEV1 values upon completion of the treatment period compared with placebo recipients (p = 0.003). Metaproterenol use decreased in both groups, but treatment differences, though not statistically significant, favored LY171883 (p = 0.089). Of patients who used at least 23 mg/wk of metaproterenol (36 inhalations) at initiation of the study, those who received LY171883 used significantly less metaproterenol than those who received placebo (p = 0.007). LY171883 was well tolerated and reduced the need for a bronchodilator drug while improving pulmonary function. Results of this study support the hypothesis that leukotrienes LTD4 and/or LTE4 may be important in the pathogenesis of asthma in humans. PMID- 2554767 TI - Autoradiographic localization of beta-adrenoceptors in asthmatic human lung. AB - The autoradiographic distribution and density of beta-adrenoceptors in human non diseased and asthmatic bronchi were investigated using [125I]iodocyanopindolol (I CYP). Analysis of the effects of the beta-adrenoceptor antagonists on I-CYP binding demonstrated that betaxolol (20 nM, beta 1-selective) had no significant effect on specific grain density in either nonasthmatic or asthmatic human bronchus, whereas ICI-118551 (20 nM, beta 2-selective) inhibited I-CYP binding by 85 +/- 9% and 89 +/- 3%, respectively. Thus, homogeneous populations of beta 2 adrenoceptors existed in bronchi from both sources. Large populations of beta adrenoceptors were localized to the bronchial epithelium, submucosal glands, and airway smooth muscle. Asthmatic bronchial tissue featured epithelial damage with exfoliated cells associated with luminal mucus plugs. A thickened basement membrane and airway smooth muscle hyperplasia were also evident. High levels of specific I-CYP binding were also detected over asthmatic bronchial smooth muscle, as assessed by autoradiography and quantitation of specific grain densities. Isoproterenol and fenoterol were 10- and 13-fold less potent, respectively, in bronchi from asthmatic lung than in those from nonasthmatic lung. However, this attenuated responsiveness to beta-adrenoceptor agonists was not caused by reduced beta-adrenoceptor density in asthmatic airways. A defect may exist in the coupling between beta-adrenoceptors and postreceptor mechanisms in severely asthmatic lung. PMID- 2554768 TI - Progressive esophagitis from acyclovir-resistant herpes simplex. Clinical roles for DNA polymerase mutants and viral heterogeneity? AB - Clinically acquired acyclovir resistance in herpes simplex has usually been associated with a deficiency in viral thymidine kinase, which, in turn, has been linked with attenuated virulence in animal models. Diminished pathogenicity in thymidine kinase-deficient isolates has been partly responsible for controversies about the clinical significance of antiviral resistance. We report on a series of resistant virus isolates from a patient who had severe, progressive esophagitis. These isolates had various thymidine kinase activities, ranging from 2.8% to 130% when compared with the activity of the isolate obtained before treatment; the resistant isolate 615 retained enzyme activity as well as neurovirulence in an encephalitis model. Plaque purification showed a heterogeneous mixture containing at least one acyclovir-resistant, foscarnet-resistant plaque isolate (615.8) fully able to phosphorylate acyclovir. The 3.3-kbp BamHI fragment containing most of the DNA polymerase gene from isolate 615.8 was purified and used to successfully transfer both acyclovir and foscarnet resistance. Acquisition of in vitro acyclovir resistance was associated with progression of clinical disease, as well as with maintenance of pathogenicity in an animal model and at least one mutation in viral DNA polymerase. Patients with herpes simplex infections that progress during acyclovir therapy should be observed for acquisition of resistance in the setting of antiviral chemotherapy; future studies should also consider the presence of heterogeneous virus populations in such patients. PMID- 2554769 TI - Adenoid cystic carcinoma of the parotid gland: a review of surgical management with reference to the facial nerve. AB - The purpose of this study was to review a large series of patients with adenoid cystic carcinoma of the parotid gland in order to develop clinical criteria for sacrifice versus selective sparing of the seventh nerve. We studied 43 patients followed over 36 years. Study patients were grouped according to the preoperative status and intraoperative management of the facial nerve. Criteria for choosing various management strategies are discussed. PMID- 2554770 TI - [Metastatic glucagonoma. A case report]. PMID- 2554771 TI - [Erythroblastopenia caused by parvovirus in siblings, revealing a hereditary spherocytosis. Apropos of a case]. AB - Following the occurrence in two siblings of parvovirus B19-related acute transient erythroblastopenia with specific IgMs, hereditary spherocytosis (HS) was diagnosed for the first time in the two patients as well as in three other members of the family. PMID- 2554772 TI - [Parvovirus infections in children with hemolytic anemia]. AB - Acute episodes of erythroblastopenia in children with chronic hemolytic anemias have been recognized for a considerable length of time. In 1981, a small virus with a single strand of DNA, parvovirus B 19, was identified as the causative agent in most such episodes. Other diseases have been ascribed to parvovirus B 19, including erythema infectiosum or fifth disease, polyarthralgia, fetal death. Schonlein-Henoch disease, and bone marrow aplasia. In acute attacks of erythroblastopenia, anemia is the most prominent manifestation, but coexistence of neutropenia and thrombopenia has been reported. Hematologic disorders last for approximately ten days. The diagnosis of recent parvovirus B 19 infection rests on detection of specific IgM antibodies, or direct visualisation of the virus by electron microscopy. More recently, detection of the viral genome using molecular hybridization techniques has been achieved. In vitro studies have confirmed the inhibiting effect of parvovirus B 19 on red cell line progenitors, particularly CFU-E, but the exact mechanism of the inhibition remains uncertain. Other diseases due to parvovirus B 19 or other parvoviruses probably remain to be discovered. PMID- 2554773 TI - [Macrophage activation syndrome linked to Epstein-Barr virus]. AB - Epstein-Barr virus infection is one of the etiologies that should be discussed in patients with macrophage activation syndrome. We report a case that is consistent with this diagnosis. The role of the Epstein-Barr virus in the etiologic diagnosis of VAHS (virus-associated hemophagocytic syndrome) is specified. In pediatric patients with VAHS, the other causes of macrophage activation should also be discussed. PMID- 2554774 TI - Treatment of facial vascular lesions with a 100-mu spot 577-nm pulsed continuous wave dye laser. AB - The present study documents the treatment and follow-up of 25 patients treated for four different types of superficial facial vascular lesions with the continuous wave, mechanically pulsed, yellow (577-nm wavelength) tunable dye laser using a 100-mu spot diameter handpiece and pulse durations of 0.05 to 0.10 s. When absorbed, this wavelength generates heat specifically within superficial blood vessels and, if delivered in a brief, intense pulse to a small, focused spot, can cause selective damage without destruction of the adjacent epidermis or surrounding dermis. The results indicate that the physician can attain an excellent clinical outcome while significantly reducing potential adverse side effects seen in treating these lesions with more conventional laser systems. PMID- 2554775 TI - Scapulectomy with latissimus dorsi musculocutaneous flap in the treatment of sarcoma of the upper back. AB - We present a patient in whom scapulectomy was performed to treat a huge malignant fibrous hystiocytoma of the back. Computed tomographic examination showed the tumor to have invaded the musculature of the upper back and about the scapula. To achieve adequate surgical excision, a major portion of the skin and soft tissue of the upper back was excised in conjunction with subtotal scapulectomy. The defect, with exposed vertebral spines and transected scapula, was covered with an ipsilateral latissimus dorsi musculocutaneous flap. Now, four years postoperatively, the patient has no evidence of recurrent disease and has good use of his arm. Total or partial excision of the scapula is rarely performed and poses unique problems. As our case demonstrates, scapulectomy (combined with musculocutaneous flap) is well tolerated, and surgeons should not be discouraged from using it when appropriate. PMID- 2554776 TI - [Super-paramagnetic iron oxide: an MRI contrast media for the reticuloendothelial system]. AB - In this preliminary study, the authors studies MR imaging in four patients with malignant hepatic tumours before and after intravenous injection of 20 mumols/kg of superparamagnetic iron oxide (SIO). They obtained spin-echo sequences (SE 500/28/4, SE 2 000/28, 56/2) before and after injection and systematically compared the results with ultrasonography and computed tomography data. In every case, superparamagnetic iron oxide (SIO) resulted in a marked decrease in T2 and consequently in the signal of healthy hepatic and splenic tissue, considerably improving the visualisation of malignant lesions which remained unchanged. If the detectability of lesions is considered to depend on the contrast obtained between tumours and healthy tissue, these preliminary results are very promising. PMID- 2554777 TI - [Isolation and in vitro and in vivo characteristics of a clone of a wild Aujeszky disease virus resistant to bromovinyldeoxyuridine]. AB - The antiviral activity of bromovinyldeoxyuridine has been studied on the wild type of pseudorabies virus. A heterogenicity of viral populations has been observed. A resistant clone presented special characteristics both in vitro (large syncytia, round cells) and in vivo (reduced pathogenicity and immunogenic power). PMID- 2554778 TI - [In vitro humoral response against the transmissible gastroenteritis virus: presentation of the antigen by the epithelial cells of the intestine]. PMID- 2554779 TI - [Expression of class II antigens by alveolar macrophages from sheep infected experimentally with the sheep pox virus]. PMID- 2554780 TI - [Expression of class II histocompatibility antigens by alveolar cells during infection by visna-maedi virus]. PMID- 2554781 TI - [BoLa and bovine leukosis]. PMID- 2554782 TI - [Antiglucocorticoid action of RU 486]. AB - RU 486 is a synthetic steroid molecule with high affinity to the glucocorticoid receptor and which exerts an antiglucocorticoid activity. In animals RU 486 constantly inhibits the various effects induced by dexamethasone in classical experimental models. Its mechanism of action is not totally elucidated: RU 486 seems to inhibit both the glucocorticoid receptor activation and the gene transcription phenomenon. The antiglucocorticoid action of RU 486 in man was first demonstrated by studying the corticotropic response: a single dose of 400 mg of RU 486, given at 0200 h, triggers a corticotropic retort as if the pituitary felt it was deprived of cortisol. RU 486 inhibits, in a dose-dependent manner, various biological responses acutely induced by dexamethasone in man: corticotropic suppression, skin blanching, circulating oesinophils drop. RU 486 suppresses the peripheral features of chronic hypercortisolism in non-pituitary dependent Cushing's syndromes. In Cushing's disease administration of RU 486 immediately induces a strong and long lasting corticotropic (and cortisol) rise, especially since the drug has a long plasma half-life. This inescapable pituitary retort is a major drawback which suggests that chronic RU 486 administration will always and inevitably induce the secretion of its own antidote...cortisol! PMID- 2554783 TI - The decrease of catalase or esterase D activity in patients with microdeletions of 11p or 13q does not increase their radiosensitivity. AB - Lymphocyte cultures from patients affected by retinoblastoma (Rb), with or without a microdeletion of chromosome 13, and Wilms tumor (WT), with a microdeletion of chromosome 11p where exposed to gamma-ray radiation during S and G2 phases. Chromatid and chromosome lesions were scored and compared to those observed in controls. No significant differences were detected, neither between patients and controls, nor between patients carrying or not a microdeletion. This lack of difference was unexpected since the genes for catalase and esterase D, also called S-formyl glutathione hydrolase, which are two detoxication enzymes, are deleted in case of microdeletion of 11p and 13q, respectively. PMID- 2554784 TI - Effects of acarbose on serum lipoproteins in healthy individuals during prolonged administration of a fiber-free formula diet. AB - The effects of the disaccharidase inhibitor acarbose on serum lipoprotein lipid concentrations were investigated in healthy subjects during prolonged feeding of a fiber-free formula diet. Acarbose was shown to decrease cholesterol and fasting triglyceride concentrations, whereas the postprandial increment of triglycerides was not diminished. The response of fasting triglycerides to acarbose treatment appeared to be related to dietary fat intake, but not to the drug-induced reduction of postprandial glucose and insulin concentrations. Both the triglyceride and the cholesterol lowering efficacy were less pronounced with a higher amount of saturated fat than with a lower intake of fat mainly composed of polyunsaturated fatty acids. The decrease in total cholesterol was shown to be a consequence of a significant reduction in low density lipoprotein (LDL) cholesterol. Since high density lipoprotein (HDL) cholesterol concentrations remained unaltered, the ratio of HDL/LDL cholesterol changed in a beneficial way. PMID- 2554785 TI - Recombinant human interferon alpha-2b enhances the radiosensitivity of small cell lung cancer in vitro. AB - We have studied the effects of human recombinant interferon alpha on the radiobiological response and the cell cycle distribution of human small cell lung cancer cell lines. In two small cell lung cancer cell lines tested so far, pretreatment with interferon alpha was found to increase the radiation sensitivity without altering the distribution of cells in the cell cycle. PMID- 2554786 TI - Differential sensitivity of tumorigenic and genetically related non-tumorigenic cells to cytotoxic polyunsaturated fatty acids. AB - A series of closely related rat brain cell lines that differ in their ability to form tumors has been used to investigate the selectivity of cytotoxic polyunsaturated fatty acids. The colony-formation ability of tumorigenic F4 cells was markedly reduced when the cells were challenged with GLA and EPA. In contrast, the non-tumorigenic revertants were less affected. All retransformed tumorigenic variants exposed to GLA were as sensitive as their parental tumorigenic cells and more sensitive than the non-tumorigenic clones. However, two out of three retransformed tumorigenic variants exposed to EPA were less sensitive than either the parental tumorigenic or non-tumorigenic clones. The addition of ferrous chloride to the culture medium increased the cytotoxicity of GLA in tumorigenic but not in non-tumorigenic variants. These results suggest that tumorigenicity per se is characterized by a high sensitivity to PUFAs exogenously administered at appropriate concentrations and that the sensitivity is fatty acid specific. PMID- 2554787 TI - Tumor progression in the human melanocytic system. AB - The isolation and routine tissue culture of melanocytic cells from normal skin, precursor nevi, primary and metastatic melanomas has allowed the experimental study of different stages of tumor progression. Characteristic differences between cultured normal melanocytes and highly malignant metastatic melanoma cells were: 1) limited life span for normal melanocytes and non-malignant nevus cells versus infinite growth for malignant melanoma cells; 2) inability to grow anchorage-independently versus high colony forming-efficiency in soft agar; 3) non-tumorigenicity versus tumorigenicity in athymic nude mice; 4) dependence on exogenous growth factors and other mitogens versus autonomous growth in protein free medium; 5) expression of melanocyte-associated antigens versus expression of melanoma-associated antigens; and 6) diploid karyotype versus non-random chromosomal abnormalities. The only major distinction found between advanced primary and metastatic melanomas was that only metastatic melanoma cells proliferated continuously in the absence of growth factors or other proteins. However, advanced primary melanoma cells could be clearly distinguished from dysplastic nevus cells by their growth behavior and growth factor requirements. Only limited information is available on the biologic, genetic, immunologic and molecular properties of dysplastic nevus cells and early (radial growth phase) primary melanoma cells but these cells appear to differ markedly from advanced primary and metastatic cells. The availability of cells from sequential steps of tumor progression in the human melanocytic system offers a unique experimental model for the study of malignant transformation. PMID- 2554788 TI - Expression of c-erbA in human hepatocellular carcinomas. AB - The expression of the oncogene c-erbA was studied in six hepatocellular carcinoma specimens and normal hepatic tissue. Total RNA isolated from these samples was analysed by Northern as well as slot blot hybridisation to a radioactive c-erbA probe. When compared to normal tissue, expression in the tumour and tissue adjacent to tumour was markedly elevated. These results suggest that overexpression of c-erbA is related to hepatocarcinogenesis. Southern blot analysis of DNA from the tumours gave no evidence of c-erbA rearrangements. PMID- 2554789 TI - Oral chemotherapy for poor risk small-cell lung cancer patients with combined idarubicin and etoposide. AB - Sixteen patients with previously untreated small-cell lung cancer, unsuitable for standard aggressive intravenous chemotherapy due to advanced age or poor performance status or very advanced disease including brain metastases or either extensive liver or marrow involvement with impaired organ function, were treated with combined oral chemotherapy including 4-demethoxydaunorubicin (IMI30, idarubicin) 30 mg/sm on day 1 and etoposide (VP16) 150 mg/sm on days 2,3,4 every 4 weeks. Out of 13 evaluable patients 1 had a complete response and 2 had a partial response with an overall objective response rate of 23% (95% confidence limits 5-53.8%). Toxicity was generally very mild. Although the compliance of this regimen is excellent, its antitumor activity seems unsatisfactory even in this category of poor-risk small-cell lung cancer patients. PMID- 2554790 TI - Benzodiazepine receptors and diazepam-binding inhibitor in human cerebral tumors. AB - Benzodiazepines can regulate neoplastic growth and immune response through specific peripheral benzodiazepine receptors. We investigated the presence of peripheral and classic central benzodiazepine receptors as well as diazepam binding inhibitor, an endogenous ligand of both types of receptors, in different human cerebral tumors. Peripheral benzodiazepine receptors were present in all the tumor types studied, whereas central benzodiazepine receptors and diazepam binding inhibitor were detectable in astrocytomas and glioblastomas and undetectable in meningiomas, neurinomas, and metastases. The role of diazepam binding inhibitor and of the different benzodiazepine receptors in neoplastic cells is still to be defined. PMID- 2554791 TI - Peripheral neuropathy in amyotrophic chorea-acanthocytosis. AB - We investigated involvement of the peripheral nervous system in 6 patients with amyotrophic chorea-acanthocytosis. Electromyographic and neurographic findings, and pathological changes as demonstrated by examination of biopsy specimens of muscle and sural nerve indicate that most patients had an axonal sensorimotor polyneuropathy with more pronounced involvement of the distal portion of the nerves. Results obtained in one patient raised the question of an anterior horn cell disorder. PMID- 2554792 TI - Protease nexin I immunostaining in Alzheimer's disease. AB - Immunohistochemical experiments showed that a small subset of neuritic plaques in brains affected by Alzheimer's disease stain positively with an antibody to the protease inhibitor protease nexin I. Nearly all plaques and tangles throughout the brain also showed a heightened avidity for a protease nexin I antibody antigen aggregate, suggesting that plaques and tangles may have a particularly high density of protease nexin I-binding sites. These observations provide further evidence for a potential disruption of protein processing in the brains of patients with Alzheimer's disease. PMID- 2554793 TI - Peripheral nerve disorders in instrumentalists. AB - I evaluated 226 instrumentalists for playing-related symptoms. There were 103 players (46%) of string instruments, 7 keyboard instrumentalists (32%), 44 players (19%) of wind instruments, and 6 percussionists (3%). The mean age was 32 years; 58% were women. Sixty-five patients (29%) had a peripheral nerve disorder; 27 had symptoms of thoracic outlet syndrome; 12 had median neuropathies (carpal tunnel syndrome in 9); 9 had ulnar neuropathies; 6 had cervical radiculopathy; 5 had digital neuropathies; 3 had cranial neuropathies; and 3 had other peripheral nerve disorders. Treatment consisted of modification of playing schedules and technique, plus physical and occupational therapy in 51 and surgical procedures in 9 patients; 5 were not treated. Follow-up at 1 to 8 years showed 74% had a satisfactory result, 14% had slight or minimal improvement, and 12% had no improvement. At the time of writing, all but 4 patients remained in the music profession. It is concluded that playing-related symptoms among instrumentalists are frequently neurological in origin and that the large majority can be helped by conservative or, on occasion, surgical means. PMID- 2554794 TI - Prevalence of Lewy bodies in Alzheimer's disease. PMID- 2554796 TI - Cell surface changes in Pseudomonas aeruginosa PAO4069 in response to treatment with 6-aminopenicillanic acid. AB - Pseudomonas aeruginosa PAO4096 was induced for beta-lactamases with 6 aminopenicillanic acid. Surface changes concomitant with beta-lactamase induction were monitored. The surface hydrophobicity of the culture increased during exposure to 6-aminopenicillanic acid. The increase was associated with a change in the distribution of the O antigen in the lipopolysaccharide of treated cells. The hydrophobicity change was reversible and partially inhibited by depressed protein synthesis. The susceptibility of induced cells to rifampin was increased transiently, suggesting increased permeability of the induced cells. PMID- 2554795 TI - Purification, toxicity, and antiendotoxin activity of polymyxin B nonapeptide. AB - Polymyxin B, a relatively toxic antibiotic, has potent endotoxin-neutralizing properties that may be beneficial as adjunctive therapy in gram-negative sepsis. Polymyxin B nonapeptide (deacylated polymyxin B) is devoid of antibiotic activity but retains the capacity to disorganize the outer membrane of gram-negative bacteria. To evaluate the potential therapeutic usefulness of this derivative, we produced purified polymyxin B nonapeptide, tested its in vivo toxicity in animals, and evaluated its in vitro antiendotoxin activity. Effectiveness as an antiendotoxin agent was assessed by examining the ability of polymyxin B nonapeptide to block the enhanced release of toxic oxygen radicals induced by lipopolysaccharide in human neutrophils (priming). In vivo, at doses of 1.5 and 3.0 mg/kg, polymyxin B nonapeptide did not exhibit the neuromuscular blocking, neurotoxic, or nephrotoxic effects that were observed with polymyxin B sulfate. Both polymyxin B and polymyxin B nonapeptide inhibited lipopolysaccharide-induced neutrophil priming in a concentration-dependent manner, but the parent compound, polymyxin B, was 63 times more effective on a weight basis. The inhibitory activity of both compounds, however, diminished rapidly when they were added after the start of the lipopolysaccharide-neutrophil incubation. We conclude that polymyxin B nonapeptide is less toxic than polymyxin B and, at the doses tested, lacks the neurotoxicity and nephrotoxicity of the parent compound. Polymyxin B nonapeptide retains the antiendotoxin activity of polymyxin B but is much less potent. The findings suggest that these compounds block an early step in the neutrophil priming process, possibly lipopolysaccharide attachment to or insertion into the neutrophil membrane. PMID- 2554797 TI - Comparison of in vivo activity of fluconazole with that of amphotericin B against Candida tropicalis, Candida glabrata, and Candida krusei. AB - Fluconazole (UK-49,858) is a new oral bis-triazole antifungal agent with demonstrated activity against Candida albicans. Because of the increasing importance of infections due to other species of Candida, we studied the efficacy of fluconazole in a rat model of established systemic candidiasis, using clinical isolates of C. tropicalis, C. glabrata, and C. Krusei. In normal rats, oral fluconazole at both 20 and 80 mg/kg per day for 7 days reduced both kidney and liver titers of C. tropicalis and C. glabrata compared with those in control animals and was only slightly inferior to amphotericin B. Both fluconazole and amphotericin B were ineffective in reducing kidney titers of C. krusei, but amphotericin B was more effective than fluconazole in reducing liver titers. Fluconazole showed no increased efficacy at the higher dose of 80 mg/kg per day compared with 20 mg/kg per day in any experiment. These results suggest that oral fluconazole may be useful in the treatment of established disseminated candidiasis caused by species other than C. albicans. Further in vivo studies are needed, however, to define minimum effective doses and length of therapy and to test additional Candida isolates. PMID- 2554798 TI - Antibacterial and immunostimulatory properties of chemotactic N-formyl peptide conjugates of ampicillin and amoxicillin. AB - N-Formyl dipeptide conjugates of ampicillin and amoxicillin related to the chemotactic peptide N-formyl-L-methionyl-L-leucyl-L-phenylalanine were synthesized and assessed for antibacterial activity and affinity for the chemotactic peptide receptor of differentiated human promyelocytic leukemia (HL 60) cells. The conjugates and parent beta-lactam antibiotics showed similar antibacterial activities against Escherichia coli and Staphylococcus aureus. The affinity of each conjugate for the chemotactic peptide receptor was determined in a competitive binding assay, using 3H-labeled N-formyl-L-methionyl-L-leucyl-L phenylalanine. All conjugates bound to the receptor, but with affinities ranging from 1/3 to 1/100 that of the tritiated substrate. There was good correlation between receptor affinity and stimulation of chemotaxis. The peptide-antibiotic conjugates also stimulated the oxidative metabolism of the HL-60 cells by inducing the production of superoxide and hydrogen peroxide as determined by Luminol- and Lucigenin-enhanced chemiluminescence. These conjugates, based on N formyl-L-methionyl-L-leucyl-L-phenylalanine, thus combine both potent antibacterial and immunostimulatory properties within the same molecule. PMID- 2554799 TI - Daptomycin (LY146032) for prevention and treatment of experimental aortic valve endocarditis in rabbits. AB - The efficacy of daptomycin (LY146032), a vancomycinlike lipopeptide antibiotic, was compared with that of antibiotics commonly in use for prevention and treatment of experimental aortic valve endocarditis in rabbits. Strains of Staphylococcus aureus. S. epidermidis, Streptococcus sanguis, and Enterococcus faecalis were used to establish endocarditis. A single 10-mg/kg dose of daptomycin and a single 25-mg/kg dose of vancomycin were both effective in prevention of endocarditis produced by strains of S. aureus and S. sanguis. Daptomycin was more effective than vancomycin for prevention of endocarditis caused by the strain of S. epidermidis. A single dose of daptomycin also was more effective in prevention of staphylococcal and enterococcal endocarditis than were single-dose regimens of cefazolin (100 mg/kg) and the combination of ampicillin (30 mg/kg) plus gentamicin (3 mg/kg), respectively. For treatment of endocarditis, daptomycin (10 mg/kg) as a single daily dose was as effective as regimens of either vancomycin or beta-lactam antibiotics for staphylococcal and enterococcal endocarditis. Daptomycin, however, was not as effective as a single daily dose of 600,000 U of procaine penicillin for endocarditis caused by the strain of S. sanguis. PMID- 2554800 TI - Effect of 2'-nor-cyclic GMP against guinea pig cytomegalovirus infection. AB - Cyclic phosphate derivative of DHPG, 2'-nor-cGMP [9-[(2-hydroxy-1,3,2 dioxaphosphorinan-5-yl)oxymethyl]-guani ne phosphate-oxide] was evaluated for activity against guinea pig cytomegalovirus (GPCMV) infection in cultured guinea pig embryo cells and in guinea pigs. By virus yield reduction and plaque reduction assays, 2'-nor-cGMP was demonstrated to be 15- to 20-fold more potent against GPCMV infection than its parental drug DHPG. The selectivity index of 2 nor-cGMP was 110, which was 10-fold higher than that of DHPG. In cultured cells, 2'-nor-cGMP attained maximal antiviral activity when added to the cells within 12 h postinfection. In the studies on GPCMV infection in guinea pigs, 2'-nor-cGMP administered subcutaneously once daily (5 mg/kg per day) for 8 days, starting 24 after virus inoculation, significantly suppressed GPCMV infectivity titers in the blood, spleen, lung, and salivary gland during acute infection (10 days postinfection) as compared with sham-treated infected animals. A greater reduction of GPCMV infectivity titers in the salivary gland was noted during chronic infection (i.e., 24 days postinfection). Clinically, splenomegaly and peripheral lymphocytosis were significantly modified as compared with the sham treated animals (P less than 0.05). The drug, administered at this dosage, was reasonably tolerated by the guinea pigs and showed clinical benefit. PMID- 2554801 TI - Hybridization analysis of three chloramphenicol resistance determinants from Clostridium perfringens and Clostridium difficile. AB - The chloramphenicol resistance determinant from a nonconjugative strain of Clostridium perfringens was cloned and shown to be expressed in Escherichia coli. Subcloning and deletion analysis localized the resistance gene, catQ, to within a 1.25-kilobase (kb) partial Sau3A fragment. The catQ gene contained internal HindII, HaeIII, and DraI restriction sites and was distinct from the catP gene, which was originally cloned (L. J. Abraham, A. J. Wales, and J. I. Rood Plasmid 14:37-46, 1985) from the conjugative C. perfringens R plasmid, pIP401. Hybridization studies were carried out with a 0.35-kb DraI-P fragment of pJIR260 as an internal catQ-specific probe and a 0.38-kb EcoRV-HinfI fragment of pJIR62 as an internal catP-specific gene probe. The results showed that the catP and catQ genes were not similar and that neither probe hybridized with cat genes from other bacterial genera. However, the catP gene was similar to the cloned catD gene from Clostridium difficile. Comparative studies with both catP and catD probes showed that these genes had significant restriction identity. We therefore suggest that these genes were derived from a common source. PMID- 2554802 TI - In vitro susceptibility studies of vancomycin-resistant Enterococcus faecalis. AB - Vancomycin resistance exhibited by Enterococcus faecalis isolates V583, V586, and V587 is described. The vancomycin MICs ranged from 32 to 64 micrograms/ml. Although resistant to vancomycin, the isolates were susceptible to teicoplanin (MIC, less than or equal to 0.5 micrograms/ml). Such a glycopeptide susceptibility profile has not been previously described for E. faecalis. Time kill studies showed that vancomycin resistance adversely affected the synergistic activity that vancomycin and aminoglycoside combinations usually demonstrate against enterococci. However, the ability to detect vancomycin resistance varied with the susceptibility testing method used. Whereas broth microdilution, broth macrodilution, and agar dilution methods detected resistance, disk-agar diffusion and the AutoMicrobic system Gram-Positive GPS-A susceptibility card (Vitek Systems Inc., Hazelwood, Mo.) did not. To detect vancomycin resistance reliably and establish the incidence of such E. faecalis isolates, adjustments in some susceptibility testing methods may be necessary. PMID- 2554803 TI - Antagonism of the cytotoxic but not antiviral effects of ara-sangivamycin by adenosine. AB - Inhibition of DNA synthesis by ara-sangivamycin was antagonized by adenosine. The 50% inhibitory concentrations increased 1.6- to 32-fold in the presence of 1.0 to 50 microM adenosine, respectively. In contrast, the inhibition of human cytomegalovirus replication by ara-sangivamycin was not antagonized by as much as 50 microM adenosine. This suggests that different enzymes were responsible for the phosphorylation of ara-sangivamycin in uninfected and infected cells. PMID- 2554804 TI - Effects of reduced cation supplement recommendations (National Committee for Clinical Laboratory Standards) on daptomycin antistaphylococcal activity. PMID- 2554805 TI - Topographical localization and characterization of microsomal glucose-6 phosphatase binding sites accessible to 4,4'-diazidostilbene 2,2'-disulfonic acid. AB - The effect of the photoactivated reagent 4,4'-diazidostilbene 2,2'-disulfonic acid (DASS) on rat liver microsomal glucose-6-phosphatase has been investigated in order to analyze the accessibility and the chemical nature of functional sites of the integral enzyme protein. The following results were obtained. (i) When native rat liver microsomes are irradiated with the photoactive reagent, the activity of glucose-6-phosphatase is progressively inhibited. However, complete reactivation is obtained by modification of the DASS-labeled microsomes with Triton X-114. (ii) Inhibition of glucose-6-phosphatase is also reversed when the DASS-labeled microsomes are treated with p-mercuribenzoate or dithiothreitol. (iii) When native microsomes are labeled with DASS an intensely fluorescent adduct is formed whose emission and excitation maximum corresponds with those obtained when cysteine or 3-mercaptopropionic acid are irradiated in the presence of the photolabile reagent. (iv) The data from fluorescence measurements show that p-mercuribenzoate and dithiothreitol reduce fluorescence labeling of the microsomes whereas Triton modification of the DASS-labeled membranes does not affect the DASS-induced fluorescence. (v) Glucose 6-phosphate hydrolysis of the partially purified glucose-6-phosphatase is also inhibited as observed with native microsomes. The DASS-induced inhibition is reversed and prevented by p mercuribenzoate; however, the partially purified enzyme cannot be reactivated by Triton X-114. (vi) When glucose-6-phosphatase is partially purified from the DASS labeled microsomes this enzyme preparation is fluorescence labeled and inhibited. From these results we conclude that DASS directly reacts with the integral phosphohydrolase mainly by chemical modification of essential sulfhydryl groups of the enzyme protein accessible from the cytoplasmic surface of the native microsomal membrane. The Triton-induced reactivation of the glucose-6-phosphatase of DASS-labeled microsomes is explained in terms of conformational changes of the integral protein elicited during modification of the surrounding membrane by detergent. PMID- 2554806 TI - Stabilization of the superoxide-generating respiratory burst oxidase of human neutrophil plasma membrane by crosslinking with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide. AB - The superoxide-generating respiratory burst oxidase (NADPH-oxidase) of neutrophil plasma membranes is known to be highly unstable. In an attempt to stabilize the enzyme, we investigated the effect of crosslinking with 1-ethyl-3-(3 dimethylaminopropyl) carbodiimide (EDC). The stability of superoxide-generating activity of plasma membrane was significantly enhanced by crosslinking. The half life (t1/2) of the activity at 37 degrees C in the absence of crosslinker was about 2 min. Crosslinking extended the t1/2 significantly. Crosslinked material exhibited a biphasic loss of activity: about half was lost in each phase with respective t1/2 values of 20 and 240 min. The lifetime of the crosslinked material at 37 degrees C was further extended (about sixfold) with 30% glycerol, and the crosslinked material was completely stable for more than 2 weeks if stored on ice. Crosslinking also stabilized the activity to the effects of high salt and detergent, both of which have inactivating effects on the oxidase. In addition, crosslinking stabilizes not only the Vm but also the Km of the enzyme, which was noted to increase upon storage in the absence of crosslinking. Unlike the native material, the crosslinked oxidase failed to be stimulated (and in fact was inhibited) by phosphatidylserine, recently reported to be an activator of the oxidase (Tamura et al. (1988) J. Biol. Chem. 263, 17,621-17,626). The crosslinked plasma membrane provides a useful stabilized system for kinetic studies. When the activated plasma membrane was treated with EDC, the stabilized oxidase could not be solubilized effectively using detergents, since greater than 95% of the activity remained with the pellet following centrifugation, perhaps due to crosslinking to the cytoskeleton. However, when the activity was first detergent solubilized, the soluble activity was also stabilized by EDC. This solubilized, crosslinked material may provide useful starting material for subsequent isolation and characterization of a stabilized active NADPH-oxidase. PMID- 2554807 TI - Effect of angiotensin II infusion in rats on Na,K-ATPase activity in renal cortical microsomal preparations. AB - Direct dose-dependent effects of angiotensin II on renal tubular sodium reabsorption have been demonstrated. Alterations in tubular sodium reabsorption may occur via modulation of renal Na,K-ATPase activity. Thus, these experiments were undertaken to ascertain whether angiotensin II could influence renal cortical Na,K-ATPase activity. Angiotensin II, 495 ng/microliters/h, or vehicle (controls) was infused for 24 h via miniosmotic pumps 48 h after rats were adrenalectomized and implanted with osmotic pumps containing 12.5 micrograms/microliters corticosterone (Treatment I) or both corticosterone and 0.2 microgram/microliter aldosterone (Treatment II), and in rats receiving 3% NaCl in their food (sodium loaded, Treatment III). Rats receiving Treatments I and III received saline to drink. Renal cortical microsomal membranes were prepared, and the effects of angiotensin II infusion on the K1/2 and Vmax for Na, K, and ATP determined. Angiotensin II infusions were associated with (i) a decrease (P less than 0.001) in the K1/2 for Na activation of Na,K-ATPase from 14 +/- 3 to 6 +/- 1 (n = 4 experiments), 16 +/- 1 to 12 +/- 1 (n = 5), and 12 +/- 3 to 7 +/- 1 (n = 5) mM (means +/- SE) for treatments I, II, and III, respectively; (ii) no changes in the K1/2 for K activation or the Km for ATP; (iii) no changes in the Vmax for Na, K, or ATP; and (iv) no change in Mg-ATPase activity. We conclude that angiotensin II infusion is associated with a decrease in the K1/2 of renal cortical Na,K-ATPase activity for sodium. This action of angiotensin II on the enzyme activity may contribute to the regulation of tubular sodium transport. PMID- 2554808 TI - Melittin inhibition of the gastric (H+ + K+) ATPase and photoaffinity labeling with [125I]azidosalicylyl melittin. AB - Melittin is a 26-amino acid amphipathic polypeptide toxin from bee venom which forms anion-selective ion channels in bilayers and biological membranes under the influence of membrane potential. Melittin has been shown to interact with a number of membrane proteins. We found that melittin inhibited K+-stimulated ATP hydrolysis by the (H+ + K+) ATPase in parietal cell apical membrane vesicles derived from histamine-stimulated rabbit gastric mucosa with a KIapp of 0.5 micron. Melittin also inhibited K+-stimulated p-nitrophenyl hydrolysis activity which is associated with the gastric (H+ + K+) ATPase in a dose-dependent manner with a KIapp of 0.95 micron. ATP-driven, K+-dependent H+ transport was inhibited over this same concentration range, even in the absence of a membrane potential. Melittin did not appear to increase the H+ leak from vesicle with preformed H+ gradients when the H+ pump was arrested by Mg2+ chelation, but all possible membrane perturbation effects were difficult to rule out. However, the data suggest that melittin exerts its inhibitory effect through interaction with the (H+ + K+) ATPase. In order to determine whether direct interactions between the (H+ + K+) ATPase and melittin occurred, a radioactive derivative of melittin, [125I]azidosalicylyl melittin, was prepared and photoreacted with sealed rabbit gastric membranes and highly purified hog gastric membrane containing the (H+ + K+) ATPase. In the purified hog preparation only a 95,000-Da band, the (H+ + K+) ATPase was labeled, while in the rabbit preparation a 95,000-Da band and one other membrane protein of 70,000 Da were labeled with this reagent. Label incorporation into the (H+ + K+) ATPase and the 70,000-Da band was greatly reduced by addition of excess unlabeled melittin, suggesting specificity of the interaction. Label incorporation occurred in the absence of ATP or added salts and was not reduced by SCH28080 (a K+ site inhibitor) suggesting that the melittin binding site was distinct from the luminal K+ site of action of SCH28080. PMID- 2554809 TI - Antibody 624H12, which detects lung cancer at early stages, recognizes a sugar sequence in the glycosphingolipid difucosylneolactonorhexaosylceramide (V3FucIII3FunLc6Cer). AB - Immunocytochemical staining of cells in sputum by rat monoclonal antibody 624H12 detects lung cancer 2 years prior to its detection by conventional diagnostic techniques. The antigen recognized by antibody 624H12 is a sugar sequence in the glycosphingolipid difucosylneolactonorhexaosylceramide (V3FucIII3FucnLc6Cer) whose structure is (formula see; text) Both fucosyl residues are required for high affinity binding by the antibody. The antigen was expressed in 35 of 45 specimens of cancer tissue from patients with early stage non small cell lung cancer. There was no correlation between antigen expression and patient survival. PMID- 2554810 TI - Hydroxyl radicals is not a significant intermediate in the vanadate-stimulated oxidation of NAD(P)H by O2. AB - The vanadate-stimulated oxidation of NADH by an enzymatic flux of O2- is inhibited by superoxide dismutase, but not by catalase. Keller et al. (1989, Free Radical Biol. Med. 6, 15-22) observed inhibition by catalase presumably because they used a commercial preparation contaminated with superoxide dismutase. Their proposal, that H2O2 and hydroxyl radical play significant roles in vanadate stimulation of NAD(P)H oxidation, may be discounted on the basis of these and of previously reported results. PMID- 2554811 TI - Protein phosphatase type 1 catalytic subunit forms nondissociable dimers. AB - Protein phosphatase type 1 is the major enzyme in skeletal muscle and liver for the dephosphorylation of Ser(P) and Thr(P) phosphoproteins. The cDNA for the catalytic subunit encodes a polypeptide of Mr 35,400 kDa, consistent with the Mr of 36,000-38,000 of the active protein purified in various laboratories. However, several investigators have found a Mr 70,000 protein for phosphatase type 1. In this report proteins of Mr 38,000 and 70,000 were resolved by Mono Q chromatography after extensive copurification from rabbit skeletal muscle. Antibodies affinity-purified against a type 1 phosphatase catalytic fragment reacted with both proteins in Western immunoblotting. Fractions from each peak were cleaved with cyanogen bromide and the major peptides were the same size by electrophoresis in gradient polyacrylamide gels. Cyanogen bromide peptides of the individual bands also were mapped by reversed-phase high-performance liquid chromatography. The purified Mr 38,000 and 70,000 proteins had identical HPLC peptide maps and also gave the same amino acid compositions after acid hydrolysis. Purified Mr 38,000 phosphatase catalytic subunit spontaneously formed a Mr 70,000 dimer that resisted usual dissociation conditions, i.e., boiling dodecyl sulfate plus 2-mercaptoethanol, but could be cleaved to about half size by various proteases, indicating that monomers were bound together near their amino or carboxy termini. Physiological changes in protein phosphatase type 1 are reflected in the amount of nondissociable dimers detected in tissue extracts. PMID- 2554813 TI - The interaction of 5,5-dimethyl-1-pyrroline-N-oxide with human myeloperoxidase and its potential impact on spin trapping of neutrophil-derived free radicals. AB - Activation of human neutrophils leads to secretion of myeloperoxidase (MPO) with resulting generation of several oxidant species including OCl-. Spin trapping techniques employing 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) are being applied increasingly to the investigation of free radical production by in vitro and in vivo experimental systems which contain neutrophils. Because such knowledge is critical to the interpretation of these data, we examined the impact of MPO and MPO-derived oxidants on DMPO spin adduct formation and stability. Addition of increasing concentrations of OCl- to DMPO yielded a number of EPR-detectable products including DMPO-OH. However, the concentration of OCl- required was in excess of that expected under physiologic conditions. Addition of purified human MPO and H2O2 to DMPO yielded EPR spectra consisting of small DMPO-OH peaks. The addition of MPO and H2O2 to preformed DMPO-OH and DMPO-CH3 resulted in rapid destruction of these spin adducts. Thus MPO/H2O2 appeared to both generate and destroy DMPO spin adducts. Neutrophils stimulated with phorbol myristate acetate or opsonized zymosan generated large DMPO-OOH and DMPO-OH peaks as well as small DMPO-CH3 peaks. Addition of the MPO inhibitor azide to the reaction mixture had no effecting on resulting DMPO-OH or DMPO-CH3 peak amplitudes but increased that of DMPO-OOH. These data suggest that MPO-derived oxidants likely have little impact on the nature of EPR spectra resulting from DMPO spin trapping of free radical species following neutrophil stimulation. Because MPO oxidants did appear to react with DMPO the ability of DMPO to protect a biologic target from in vitro MPO injury was examined. DMPO (greater than 10 mM) significantly decreased MPO/H2O2/Cl- -mediated erythrocyte hemolysis as assessed by 51Cr release. The experimental and/or pharmacologic implications of this observation require further study. PMID- 2554814 TI - A comparison of cobalt(II) and iron(II) hydroxyl and superoxide free radical formation. AB - We have employed the electron spin resonance spin-trapping technique to study the reaction of Co(II) with hydrogen peroxide in a chemical system and in a microsomal system. In both cases, we employed the spin trap 5,5-dimethyl-1 pyrroline N-oxide (DMPO) and were able to detect the formation of DMPO/.OH and DMPO/.OOH. DMPO/.OOH was the predominant radical adduct formed in the chemical system, while the two adducts were of similar concentrations in the microsomal system. The formation of both of these adducts in either reaction system was inhibited by the addition of superoxide dismutase or catalase, and by chelating the cobalt with either ethylenediaminetetraacetic acid (EDTA) or diethylenetriaminepentaacetic acid (DTPA). The incorporation of the hydroxyl radical scavengers ethanol, formate, benzoate, or mannitol inhibited the formation of DMPO/.OH in both systems. We also repeated the study using Fe(II) in place of Co(II). In contrast to the Co(II) results, Fe(II) reacted with hydrogen peroxide to yield only DMPO/.OH, and this adduct formation was relatively insensitive to the presence of added superoxide dismutase. In addition, Fe(II) mediated DMPO/.OH formation increased when the iron was chelated to either EDTA or DTPA rather than being inhibited as for Co(II). Thus, we propose that Co(II) does not react with hydrogen peroxide by the classical Fenton reaction at physiological pH values. PMID- 2554812 TI - The role of sterol carrier protein 2 in stimulation of steroidogenesis in rat adrenal mitochondria by adrenal cytosol. AB - Cholesterol side-chain cleavage (CSCC) in isolated rat adrenal mitochondria is enhanced by prior corticotropin (ACTH) stimulation in vivo (8-fold). Part of this stimulation is retained in vitro by addition of cytosol from ACTH-stimulated adrenals to mitochondria from unstimulated rats (2.5- to 6-fold). In vivo cycloheximide (CX) treatment fully inhibits the in vivo response and resolves the in vitro cytosolic stimulation into components: (i) ACTH-sensitive, CX-sensitive; (ii) ACTH-sensitive, CX-insensitive; and (iii) ACTH-insensitive, CX-insensitive. These components contribute approximately equally to stimulation by ACTH cytosol. Components (i) and (iii) most probably correspond to previously identified cytosolic constituents steroidogenesis activator peptide and sterol carrier protein 2 (SCP2). SCP2, as assayed by radioimmunoassay or ability to stimulate 7 dehydrocholesterol reductase, was not elevated in adrenal cytosol or other subcellular fractions by ACTH treatment. Complete removal of SCP2 from cytosol by treatment with anti-SCP2 IgG decreased cytosolic stimulatory activity by an increment that was independent of ACTH or CX treatment. Addition of an amount of SCP2, equivalent to that present in cytosol, restored activity to SCP2-depleted cytosol but had no effect alone or when added with intact cytosol, suggesting the presence of a factor in cytosol that potentiates SCP2 action. Pure hepatic SCP2 stimulated CX mitochondrial CSCC 1.5- to 2-fold (EC50 0.7 microM) but was five times less potent than SCP2 in adrenal cytosol. Two pools of reactive cholesterol were distinguished in these preparations characterized, respectively, by succinate-supported activity and by additional isocitrate-supported activity. ACTH cytosol and SCP2 each stimulated cholesterol availability to a fraction of mitochondrial P450scc that was reduced by succinate but failed to stimulate availability to additional P450scc reduced only by isocitrate. PMID- 2554816 TI - [Intraarterial chemotherapy of experimentally metastasized liver carcinoma with anticancer emulsion containing surface-active agent]. AB - The w/o Emulsion (w/o) of 0.3% adriamycin (ADR) in the mixture of a water-soluble contrast medium, 60% urografin, and an oily contrast medium, lipiodol, at the ratio of 1 : 4 was combined with 0.5% lecithin. This combination could stabilize the micelles and make the emulsion desirably slow releasing form. ADR emulsion was injected into the hepatic artery of rabbit with metastatic liver carcinoma 7 days after intraportal transplantation of VX2 tumor. The examination at day 5 revealed good accumulation and retention of ADR in tumor. This emulsion was histologically confirmed to be effective in suppressing the growth of metastatic lesion, and it was considered that this ADR emulsion would have a great possibility for clinical application in the treatment of hepatic metastasis of gastrointestinal carcinomas. PMID- 2554815 TI - [Prevention of renal toxicity of cisplatin by administration of bismuth subnitrate]. AB - The effect of pretreatment with bismuth subnitrate (BSN) for prevention of the renal toxicity of cisplatin (CDDP) was examined in 44 patients with lung cancer (43 non-small cell and one small cell lung cancer). In non-small cell lung cancer cases, the effect of the antitumor activity of chemotherapeutic drugs was observed in 62% of patients pretreated with BSN, and 42% in the group without pretreatment with BSN. No antitumoral activity of chemotherapeutic drugs was suppressed by treatment with BSN. In the group without pretreatment of BSN, serum creatinine and BUN were in proportion to the number of administrations of chemotherapeutic drugs. On the other hand, no renal toxicity was shown in the group with pretreatment by BSN. No protective effect against myelosuppression with pretreatment by BSN was demonstrated, perhaps because of the influence of anti-cancer drugs apart from CDDP. PMID- 2554817 TI - [A case of intra-arterial infusion chemotherapy in small cell lung cancer with liver metastases]. AB - A 53 year old male was admitted with cough, chest pain and bloody sputa for one month. His admission chest radiography revealed a tumor shadow in right hilus. The patient was diagnosed as small cell lung cancer (oat cell type) by transbronchial biopsy. Clinical staging was IIIA and performance status was 1. The patient was treated by combined chemotherapy (CPA, ADM and VCR) for 3 courses and chest irradiation (5,000 rad). After such therapy, the primary site was regressive until 2 months prior to death. One month after irradiation, abdominal CT showed multiple liver metastases. Though CDDP 100 mg/body and etoposide 100 mg/body X5 were administered systemically, improvement of metastases of the liver was not revealed by abdominal CT. However, after hepatic arterial infusion of ADM (10 mg/body) suspended in a lipiodol (3 ml/body) and CDDP (100 mg/body) was performed, liver metastases were remarkably regressive by abdominal CT. The patient died of a systemic relapse about 14 months after liver involvement. PMID- 2554818 TI - Cutaneous leishmaniasis. Treatment with itraconazole. AB - The chemotherapy of leishmaniasis is still far from satisfactory. Itraconazole, one of the most recent azole antifungal agents, appears to be well tolerated in man. Two patients with cutaneous leishmaniasis (Leishmania tropica), contracted in Saudi Arabia, were treated orally for 2 months with itraconazole (100 mg/d). Both recovered without side effects or abnormalities in their main biologic parameters. PMID- 2554819 TI - Successful treatment of progressive acyclovir-resistant herpes simplex virus using intravenous foscarnet in a patient with the acquired immunodeficiency syndrome. AB - We report a case of orofacial herpes simplex virus (HSV) infection that was progressive despite multiple courses of acyclovir sodium in a patient with the acquired immunodeficiency syndrome. The viral isolate was shown to be resistant to acyclovir in vitro, but proved susceptible to vidarabine and foscarnet sodium (trisodium phosphonoformate). The patient failed to respond to a 2-week course of intravenous vidarabine. However, rapid improvement in the orofacial lesion occurred with intravenous foscarnet. Most HSV isolates that are resistant to acyclovir are spontaneous mutants partially or completely lacking in thymidine kinase. Because foscarnet is a direct inhibitor of HSV DNA polymerase, this compound is expected to have efficacy against acyclovir-resistant strains. This report documents successful treatment of clinically significant HSV with intravenous administration of foscarnet, suggesting that further study is indicated. PMID- 2554820 TI - Human papillomavirus associated with keratoacanthoma. PMID- 2554821 TI - Prevalence of HPV 7 papillomas in the oral mucosa and facial skin of patients with human immunodeficiency virus. PMID- 2554822 TI - Routine immunisation of preterm infants. AB - Fifty one preterm infants (26-36 weeks' gestation) were enrolled in a study of their immunological responses to diphtheria, tetanus, pertussis, and polio antigens eight to 12 weeks after their primary courses had been completed. Samples from 21 infants born at full term were also analysed. Many infants were able to start immunisation at 3 months of age. Premature infants who are immunised as soon as possible after 3 months of age develop adequate antibody responses. PMID- 2554823 TI - Neonatal urological ultrasound. PMID- 2554824 TI - Quality of community drinking water and the occurrence of spontaneous abortion. AB - To investigate the relationship between community drinking water quality and spontaneous abortion, we compared trace element levels in the drinking water of 286 women having a spontaneous abortion through 27 wk gestation with that of 1,391 women having livebirths. Trace element levels were gathered from routine analyses of public tap water supplies from the communities where the women resided during pregnancy. After adjustment for potential confounders, an increase in the frequency of spontaneous abortion was associated with detectable levels of mercury; high levels of arsenic, potassium, and silica; moderately hard water, and surface water. In contrast, a decrease in the frequency of spontaneous abortion was associated with high levels of alkalinity and sulfate, and any detectable level of nitrate. These results require further corroboration because there is a paucity of data investigating this issue. PMID- 2554825 TI - [Microscopic innervation of the spermatic ducts and testis. II. Ductus epididymis]. AB - The microscopic innervation of the ductus epididymis in the rat has been studied both in light and electron microscope. The nerve fibres form perivascular, intramuscular and subepithelial plexuses. Occasionally sensory formations were observed in the adventitia of the cauda epididymis. The innervation is more abundant in the cauda epididymis than in the other segments (caput and corpus). The varicosities and endings of nerve fibres have synaptic vesicles of different sizes and electron characteristics. Some of the varicosities were close to the muscle cells and without glial sheath. The epithelial cells were not directly innervated. PMID- 2554826 TI - Autoantibodies to recombinant lipocortin-1 in rheumatoid arthritis and systemic lupus erythematosus. AB - Corticosteroids may mediate some of their anti-inflammatory effects via induction of a specific 38 kD protein, lipocortin-1. Autoantibodies to lipocortin-1 were measured by enzyme linked immunosorbent assay (ELISA) in 90 healthy subjects and in 63 patients with rheumatoid arthritis (RA), 36 with systemic lupus erythematosus (SLE), 26 with polymyalgia rheumatica, and 13 with chronic airways disease. Sixteen patients with RA receiving prolonged, high steroid doses (prednisolone greater than 7.5 mg/day) had raised IgM antilipocortin-1 levels, while 19 patients with RA untreated with steroids had normal levels. This association was independent of disease activity. In SLE, raised antilipocortin-1 levels were associated with active disease and were independent of steroid treatment. Antilipocortin-1 antibody levels were not raised in patients with polymyalgia rheumatica and chronic airways disease. Thus steroid treatment alone appears insufficient to induce antilipocortin-1 antibodies, unless an underlying autoimmune state is also present. In RA, antilipocortin-1 antibodies may impair anti-inflammatory actions of steroids and render some patients 'steroid resistant'. PMID- 2554827 TI - Natural history of in situ breast cancer in a defined population. AB - The entire experience of in situ breast cancer in Alberta from 1953 to 1984 was examined. Of 243 patients coded, 226 were available for review by a panel of three pathologists. In 149 cases the diagnosis of in situ disease was confirmed. One hundred and eight patients had 109 ductal carcinomas in situ, 38 patients had lobular carcinomas in situ, with 3 patients having both. A multitude of treatments was used, ranging from local excision to radical mastectomy. Survival at a mean of 6 years follow-up was equal in all groups, with only two patients with a confirmed diagnosis of ductal carcinoma in situ dying from clinically suspected systemic disease. In patients treated by local excision, ipsilateral cancers were seen in 12% of ductal carcinoma in situ patients who had local excision and 13% of patients with lobular carcinoma in situ. Contralateral metachronous invasive cancers were seen in 6% of ductal carcinoma in situ patients and 3% of lobular carcinoma in situ patients. No lymph node involvement was seen in any of these patients, either with prophylactic dissection or in follow-up. The conclusion reached was that both in situ lesions are similar in their clinical course. Lymph node dissection is not necessary. Pathologic review is critical for accurate studies, with a change in diagnosis of 36% of diagnoses. Treatment does not appear to affect prognosis. The most appropriate treatment needs to be determined in prospective randomized trials. PMID- 2554828 TI - Longer survival after resection of non-small cell lung cancer in Japanese women. AB - From 1974 to 1988, 492 patients (361 male and 131 female) with non-small cell lung cancer underwent pulmonary resection at our department. A retrospective study was done with special reference to the sex of the patients. There was no sex-related association with age distribution, operation, pathological T or N status, or staging. On the other hand, 77.9% of women had an adenocarcinoma and only 44.6% of men had this type of lesion. This was considered at least partly a result of sex difference in smoking habits. Women survived for a significantly longer time (p = 0.0036), and this difference could not be explained by differences in histological type. Presumably, hormonal factor(s) may contribute to the prognosis. PMID- 2554829 TI - Pulmonary artery versus left ventricular venting: a radioisotope study of left ventricular function. AB - Radionuclide measurements of ejection fraction were used to assess immediate and late postoperative ventricular function after the use of either a pulmonary artery vent (group A) or a left ventricular vent (group B) in 20 patients undergoing aortic valve replacement for pure aortic stenosis. Ten patients were included in each group and anesthetic techniques, patient management, and septal temperatures were similar in all cases. No significant difference was found between the preoperative and immediate or 6-week postoperative ejection fractions, either taken overall or between the two groups (p greater than 0.05; Student's t test). No correlation was found between cross-clamp time, bypass time, or the occurrence of ventricular fibrillation and the immediate postoperative ejection fraction (p greater than 0.05; Student's t test). There was no significant difference in the incidence of ventricular fibrillation after each type of vent had been used (chi 1(0) = 3.32; p greater than 0.05). We did not demonstrate any abnormalities in regional wall motion associated with apical insertion of a left ventricular vent, and conclude that pulmonary artery and left ventricular vents are equally satisfactory in terms of postoperative ventricular performance. PMID- 2554830 TI - Blood transfusion and lung cancer recurrence. PMID- 2554831 TI - A preliminary report of the short-term effect of carbonated beverage consumption on calcium metabolism in normal women. AB - A variety of nutritional factors influence the bioavailability of calcium and increase a woman's risk of osteoporosis. Eight healthy women completed an 8-week metabolic study designed to investigate the effect of nonalcoholic carbonated beverage consumption on calcium metabolism. Compared with women receiving a control diet, women consuming a diet high in nonalcoholic carbonated beverages demonstrated similar mean serum levels of calcium, ionized calcium, phosphorus, alkaline phosphatase, parathyroid hormone, 1,25-dihydroxyvitamin D3, and osteocalcin. Twenty-four-hour urine volume, creatinine clearance, calcium creatinine ratio, and phosphorus-creatinine ratio were similar during consumption of the diet high in nonalcoholic carbonated beverages and the control diet. Twenty-four-hour cyclic adenosine monophosphate-creatinine ratio was significantly lower in women consuming the diet high in nonalcoholic carbonated beverage compared with women receiving the control diet (342 +/- 27.4 nmol/mmol vs 409 +/- 22.1 nmol/mmol). Consumption of a diet high in nonalcoholic carbonated beverages on a short-term basis does not appear to affect adversely the serum or urinary markers of calcium metabolism. PMID- 2554832 TI - Three opportunistic infections associated with ectopic corticotropin syndrome. AB - Cytomegalovirus pneumonia, Pneumocystis carinii pneumonia, and pulmonary and disseminated aspergillosis occurred simultaneously in a 66-year-old white man with oat cell carcinoma and ectopic corticotropin production. Hypokalemia, a recent normal chest roentgenogram, and a large left adrenal mass on a computed tomographic scan confused the initial clinical evaluation. The aspergillosis proved fulminant and lethal. PMID- 2554833 TI - [Sequential study of the plasmatic atrial natriuretic factor and the urinary excretion of an ouabain displacing factor and of dopamine in normotensive pregnant women before and after labor]. AB - Estimation of urinary excretion of a ouabain displacing factor (ODF) and dopamine was carried out immediately before delivery, 7 days and 70-90 days after delivery in 12 normotensive pregnant women. Simultaneous estimation of plasma 99-126 atrial natriuretic factor (ANF), plasma renin activity (PRA) and plasma aldosterone were also undertaken. The data were compared with those obtained in a non pregnant normotensive group of women (n = 14) and a group of pregnant normotensive women in the early third trimester (n = 14). Urinary ODF and dopamine were significantly higher in the early third trimester when compared with non pregnant women but immediately before delivery, ODF excretion had fallen below non pregnant values and dopamine excretion had dropped to control values. Both remained low after delivery. Plasma ANF was higher in pregnant women when compared with non pregnant controls and remained high just before delivery and 7 and 70-90 days after delivery. PRA and plasma aldosterone were higher during pregnancy and had fallen to non pregnant values 7 days post-partum. It is concluded that there is considerable discrepancy in the evolution of natriuretic and antinatriuretic factors before and after delivery and that the drop of PRA and aldosterone by 7 days post-partum, contrasting with the unchanged high values of ANF, may contribute to negative sodium balance after delivery. PMID- 2554834 TI - [Alpha 2-adrenergic receptors and arterial pressure]. AB - The aim of this work was to investigate the influence of blood pressure levels on human platelet alpha 2-adrenergic receptivity. The study was carried out on 12 mild essential hypertensive patients and 7 normotensive parkinsonians with orthostatic hypotension. Alpha 2-adrenoceptors number and affinity were determined by 3H-yohimbine binding, plasma catecholamines were measured by HPLC and adrenaline-induced platelet aggregation by turbidimetry. Results obtained were compared with those of two groups of 12 normotensive control subjects. In hypertensive patients, both platelet alpha 2-adrenoceptors (139 +/- 6 vs 176 +/- 18 fmol/mg protein) and velocity of adrenaline-induced platelet aggregation were decreased whereas plasma adrenaline and noradrenaline remained unchanged. In patients with orthostatic hypotension, there was an increased number of platelet alpha 2-adrenoceptors (313 +/- 52 vs 168 +/- 8 fmol/mg protein) associated with a significant decrease in plasma noradrenaline (62 +/- 11 vs 190 +/- 25 pg/ml). In none of the two groups of patients there was any change in receptor affinity for 3H-yohimbine. These results indicate that human platelet alpha 2-adrenoceptors levels are related to blood pressure values. Moreover, up-regulation in orthostatic hypotension and lack of down-regulation in essential hypertension suggest that only sustained abnormal plasma noradrenaline levels could allow the development of alpha 2-adrenoceptors regulatory mechanisms. These variations can represent tentative compensatory mechanisms for normalization of blood pressure levels. PMID- 2554835 TI - [Visco-elastic characteristics of the aorta in conscious dogs in a model of experimental calcinosis]. AB - Effects of experimental calcinosis induced by daily overdose of 500.000 IU Vit D3 during 10 days were studied in 6 mongrel conscious dogs chronically instrumented with intra-aortic Konigsberg microtransducer and two ultrasonic piezo-electric crystals diametrically opposed in the adventitia of the descending thoracic aorta. Simultaneous recording of instantaneous aortic pressure and diameter waves in basal state and during transient acute hypertension induced by intravenous angiotensin bolus (0.1 microgram/kg) allowed to obtain the pressure (y) diameter (x) relationship of the aorta according to an exponential regression: P = expo (beta D + A), with a highly significant correlation coefficient in all animals (r greater than 0.99). (table; see text) Anatomopathological studies of aortas revealed abnormal calcium deposition, ruptures of elastic fibers and disorganization of collagen. Thus, a striking decrease in aortic rigidity is observed after calcinosis in relation with structural changes of elastic materials and responsible in part for a reduction in pulsatile pressure; moreover this unexpected phenomena might represent an initiative process of development of aortic aneurysms. PMID- 2554836 TI - [Evidence for a glomerular receptor for bradykinin in rats. Effect of sodium intake on density and affinity of the receptor]. AB - Bradykinin (BK) have been involved in a lot of pharmacological and biological effects including natriuresis, vasodilatation, inflammation and pain mediation. All these potent effects of BK are presumably mediated via one or more specific receptors which have been classified in two types named B1 and B2 receptors. In the kidney, specific binding have been reported successively in cortical epithelial membranes, in renomedullary interstitial cells and in cortical collecting tubules. Furthermore, since a large number of vasoactive compounds have been shown to regulate renal glomerular hemodynamics, we examined the binding of BK in rat glomerular membranes and the effect of variable salt diet on the density (Bmax) and the dissociation constant (KD) of this binding. Incubation of a radiolabeled bradykinin analog, [125I]-Tyr8-BK with a crude membrane preparation obtained from isolated rat glomeruli revealed a time dependent binding. The binding was saturable, reversible and was a linear function of protein membrane concentration. The radiolabeled Tyr8-BK bound to a single class of binding sites with an equilibrium dissociation constant (KD) of 2.75 +/- 0.7 nM and a density (Bmax) of 32.1 +/- 5.2 fmol/mg protein. [125I]-Tyr8-BK binding was reversed by bradykinin (Ki = 0.4 10(-9) M) and by other kinin analogs. However, Des-Arg9-BK had no effect on binding of the radiolabelled BK. These results are consistent with the presence of a B2-kinin like receptor in rat glomeruli. Low salt diet (during one month) and water deprivation (during 4 days) induced a decrease in the density of glomerular BK like receptors respectively (Bmax = 12.45 +/- 1.3 and 13.25 +/- 1.17 fmol/mg protein).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554837 TI - [Effect of beta-adrenergic receptors blockade on auricular natriuretic factor, aldosterone and renine blood activity during physical exercise]. AB - Physical exercise stimulates the renin-angiotensin-aldosterone system. However several factors affect the control of mineralocorticoid secretion. In this study, eight healthy volunteers performed maximal exercise on cycle ergometer after being pretreated for 3 days with placebo (P) or with a non selective beta-blocker (B) (pindolol 15 mg/day). Plasma reinin activity (PRA), aldosterone (ALD), atrial natriuretic factor (ANF), and kalemia (K+) were measured at rest (R) and during exercise until exhaustion (E). (table; see text) These results confirm the role of beta-adrenoceptor activation in the increased PRA during exercise. It appears an exercise-induced increase in plasma ANF which was more elevated in subjects treated with pindolol, but which had no inhibitory effect on ALD secretion in theses conditions. K+ rose during exercise and this hyperkalemia tended to be higher with a beta-blocker. It is suggested that K+ elevation counterbalance both PRA decrease and ANF increase to be responsible for the absence of change in plasma ALD during beta-blockade. PMID- 2554838 TI - [Homologous and heterologous regulation of atrial natriuretic factor receptors in smooth muscle cells]. AB - Two subtypes of atrial natriuretic factor (ANF) receptors are present in vascular smooth muscle cells: B(biologically active) receptors coupled to a guanylate cyclase and C (clearance) receptors (95 per cent of the total number of ANF binding sites) non coupled to any identified second messenger system. We compared the homologous receptor regulation induced by ANF to the heterologous one elicited by angiotensin II (Ag II). Binding of (3-[125I]iodotyrosyl) rat ANF and cGMP production stimulated by ANF were measured after 18 hours preincubation of rat cultured vascular smooth muscle cells (10(6) cells/dish) at 37 degrees C with ANF or Ag II. The hormones (10 nM) decreased to the same extent the total apparent number of ANF binding sites (control: 208 +/- 25 fmol/10(6); ANF : 82 +/ 20 fmol/10(6) cells; Ag II : 90 +/- 9 fmol/10(6) cells) The diminution of the number of ANF binding sites induced by ANF exposure was reversed by 85 per cent following 10 minutes treatment of the cells with 10 mM AcOH. Moreover, treatment with ANF (10 nM) led to a diminution of cGMP stimulation induced by ANF, this effect being still present after washing the cells with 10 mM AcOH. In contrast, diminution of ANF building sites consecutive to Ag II exposure was not affected by AcOH treatment and a potentiation of cGMP production elicited by ANF was observed. These results suggest that, in rat vascular smooth muscle cells, B receptors are sensitive to homologous down regulation and C receptors are sensitive to heterologous regulation by Ag II. PMID- 2554839 TI - [Decrease in the regulatory capacity of the calcium pump in a subgroup of essential hypertensive patients]. AB - We developed a new flux technique, useful for the clinical investigation of Ca2+ pump kinetics in intact erythrocytes. The initial rate of Sr2+ efflux (mediated by the Ca2+ pump) was studied as a function of steady state erythrocyte Sr2+ and Ca2+ contents in 22 Caucasian essential hypertensive patients (8 females, 14 males), aged 39-66 years (mean 51) and compared with 20 normotensive control subjects (10 females, 10 males), aged 22-57 years (mean 41). Kinetic analysis of the data (by using a two-sites model) allowed the determination of the apparent dissociation constants for internal Ca2+ (KCa) and for internal Sr2+ (KSr) and the maximal rate of Sr2+ efflux (Vmax). Mean values of these kinetic parameters were slightly increased in the hypertensive population. However, only the increase in KCa reached statistical significance (73 +/- 7 vs 55 +/- 3 mumol/l.cells, Mann-Whitney U test; p = 0.042). Individual analysis of the data showed that 6 essential hypertensives had a KCa higher than the upper normal limit (95 p. 100 confidence limit) of the normotensive group. In addition, mean values of Vmax and KSr were also significantly higher in these six essential hypertensives. In conclusion, about 25 p. 100 of the hypertensive patients had a decreased apparent affinity of the Ca2+ pump for internal Ca2+, which appears to be compensated (in the basal state) by an increased maximal pump rate. A similar abnormality in vascular smooth muscle cells may induce increased contractility by transitory cell Ca2+ retention after the opening of Ca2+ channels. PMID- 2554840 TI - [Therapeutic response to canrenone of patients with essential hypertension as a function of sodium transport anomalies and ouabain sensitivity of erythrocytes]. AB - The presence of Na+ transport abnormalities (decreased affinity of the Na+/K+ pump or the Na+, K+ cotransport for internal Na+, increased Na+:Li+ countertransport, increased Na+ leak), Na+ content, Na+/K+ pump activity and sensitivity to ouabain were investigated in erythrocytes from 13 patients with essential hypertension. According to the presence or absence of Na+ transport abnormalities, the patients were divided into two groups: TrNa(+) (n = 9) and TrNa(-) (n = 4) respectively. Compared with TrNa(-) patients, TrNa(+) patients were characterized by: (i) a higher arterial pressure (131.4 +/- 11.8 vs 110.0 +/ 13.2 mmHg, p less than 0.05), (ii) an increased erythrocyte Na+ content (8.9 +/- 1.0 vs 6.3 +/- 0.8 mmol/l.cells, p less than 0.01) associated with (iii) a decreased rate constant of Na+/K+ pump activity (235 +/- 26 vs 309 +/- 45 h-1, p less than 0.05) and (iv) a higher sensitivity to ouabain (0.76 +/- 0.23 vs 1.12 +/- 0.26 microM, p less than 0.05). Oral administration of canrenone 50 mg per day during 7 weeks decreased mean arterial pressure by 10-30 mmHg in 6 out of the 9 TrNa(+) patients. Conversely, it decreased mean arterial pressure in only one out of the 4 TrNa(-) patients. The hypotensive effect of canrenone in TrNa(+) patients was not associated with normalization of their Na+/K+ pump activity. Canrenone did not modify the sensitivity to ouabain of either the TrNa(+) or the TrNa(-) patients. Before treatment, acute injection of ouabain provoked an inhibition of the erythrocyte Na+/K+ pump, without any change in Na+ content.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554841 TI - Hypercortisolism among socially subordinate wild baboons originates at the CNS level. AB - Recent studies suggest that the hypercortisolism and dexamathasone resistance of depression arise, at least in part, at the level of the brain, ie, cortisol releasing factor (CRF) and/or other corticotropin-secretagogues are hypersecreted. This article suggests a similar cause of the hypercortisolism of social subordinance. Two troops of wild olive baboons, living freely in the Serengeti Ecosystem of East Africa, have been under long-term study. Consistently, in stable dominance hierachies, subordinate males are hypercortisolemic relative to dominant animals. Furthermore, hypercortisolemic males are dexamethasone resistant. There are no rank-related difference in cortisol clearance or adrenal sensitivity to corticotropin, suggesting a pituitary and/or neural locus of the hypercortisolism. Subordinate males were shown to secrete less corticotropin in response to a CRF-challenge than did dominant males. Following the logic used in similar studies with depressives, if subordinate males were hypercortisolemic despite decreased pituitary sensitivity to CRF, then this implies that the hyperactivity of the adrenocortical axis is driven at the level of the brain. Furthermore, subordinate males were hyporesponsive to CRF after administration of metyrapone, which blocks cortisol secretion and disinhibits the pituitary from feedback inhibition. Thus, the pituitary appears to have lost sensitivity to CRF itself in these low-ranking males. These observations are interpreted in light of behavioral data suggesting that these subordinate males are under sustained social stress. PMID- 2554842 TI - Cystosarcoma phylloides of the breast and its mimics. An immunohistochemical and ultrastructural study. AB - Cystosarcoma phylloides of the breast is a tumor composed of breast ducts and a cellular stromal component that can be benign or malignant. The origin of the stromal cells is controversial. We undertook an immunohistochemical and ultrastructural study of 11 cases of cystosarcoma phylloides to assess the histogenesis of the stromal component. By light microscopy, 4 were diagnosed as benign, and 7 were diagnosed as malignant. Antibodies to vimentin, desmin, actin, high- and low-molecular-weight keratins, and S100 protein were used for immunohistochemical staining. In the 4 benign cases of cystosarcoma phylloides, the stromal cells stained positively only for vimentin. In the malignant tumors, the spindle cell component stained for vimentin in all the cases. In addition, the malignant stromal cells coexpressed desmin in two cases and keratin and S 100 protein in another case. By electron microscopy the stromal component in the benign case and in two of five malignant cases was composed of a mixture of fibroblasts and myofibroblasts. The entire neoplastic stroma in two other malignant cases showed features of smooth-muscle differentiation, whereas in another case all the stromal cells showed myoepithelial differentiation. Thus, in benign and malignant cystosarcoma phylloides, the stromal component consists of a mixture of fibroblasts and myofibroblasts. Leiomyosarcomas and myoepitheliomas can mimic malignant cystosarcoma phylloides, but immunohistochemistry and electron microscopy can differentiate these entities. This is important since their biologic behavior is different. PMID- 2554843 TI - Analysis of human immunodeficiency virus and cytomegalovirus infection by polymerase chain reaction in the acquired immunodeficiency syndrome. An autopsy study. AB - Fixed autopsy tissues from nine patients infected with the human immunodeficiency virus (HIV) were assayed for the presence of HIV provirus and/or cytomegalovirus by the polymerase chain reaction. The HIV provirus was detected in lymphoid tissues from all nine patients. With nonlymphoid tissue, HIV was detected 32% (9/28) of tissues with chronic inflammation and in 7% (4/57) of tissues without chronic inflammation. Cytomegalovirus was detected in tissues from six of the nine patients, often in the absence of inclusions, and was most often detected in the adrenal gland and lung. Widely disseminated cytomegalovirus infection was present in three patients with characteristic cytomegalovirus inclusions. A similar pattern of widely disseminated HIV was not identified. These studies correlate histologic features with the presence of specific viral sequences in HIV-infected patients. PMID- 2554844 TI - Fungemia caused by an amphotericin B-resistant isolate of Sporothrix schenckii. Successful treatment with itraconazole. AB - A 58-year-old, alcoholic, diabetic man presented with multiple, ulcerated skin lesions and polymicrobial septicemia. Sporothrix schenckii was recovered from blood cultures and was resistant to amphotericin B by in vitro testing. Amphotericin B therapy failed, but the patient responded dramatically to itraconazole therapy, only to relapse 3 months after therapy was stopped. Reinstitution of itraconazole therapy has produced another dramatic response. This report is noteworthy for three reasons. First, to our knowledge, it represents only the second reported instance of fungemia with S schenckii that responded to medical therapy. Second, it illustrates that in vitro antifungal susceptibility tests may predict clinical infection response to drug therapy. Third, it suggests that itraconazole has significant promise in treating systemic sporotrichosis. PMID- 2554845 TI - Disseminated cytomegalovirus infection presenting with acalculous cholecystitis and acute pancreatitis. AB - Disseminated cytomegalovirus (CMV) infection occurs predominantly in immunocompromised hosts. Symptomatic CMV cholecystitis and pancreatitis are quite rare, and, to our knowledge, there are no reports of these occurring simultaneously. We describe a patient with a history of chronic myelogenous leukemia (treated with chemotherapy) who presented with recurrent unexplained fevers and an acute abdomen. At surgery, cholecystitis and pancreatitis were found, and a cholecystectomy was performed. The patient developed disseminated intravascular coagulation, eventual sepsis, and multiorgan failure. At autopsy, widespread disseminated CMV infection was found, with CMV-associated foci of acute inflammation and necrosis in the pancreas and in the surgically resected gallbladder. A review of our autopsy files revealed only one renal transplant patient with CMV inclusions and chronic pancreatitis. No pancreatitis was seen in 27 patients with acquired immunodeficiency syndrome. Cytomegalovirus should be considered as a possible cause of pancreatitis and cholecystitis in immunocompromised patients. PMID- 2554846 TI - Federal guidelines for marijuana screening should have lower cutoff levels. A comparison of results from immunoassays and gas chromatography-mass spectrometry. AB - Federal guidelines for urine marijuana testing use a screening cutoff of 100 ng/mL and a confirming cutoff of 15 ng/mL. We tested 75 urine samples with two different immunoassays with cutoff points of 100 and 20 ng/mL. The same samples were also analyzed with gas chromatography-mass spectrometry. At the government recommended cutoff levels for screening and confirming, test sensitivity was 47% and specificity was 91%. With a screening cutoff of 20 ng/mL confirmed at 5 ng/mL, test sensitivity was 88% and specificity was 94%. We suggest adoption of these lower cutoff levels. PMID- 2554847 TI - The prevalence of peripheral neuropathy in patients with anorexia nervosa. AB - A prospective, controlled study was conducted to determine the prevalence of peripheral neuropathy (PN) in patients with anorexia nervosa (AN). Fifty-one patients (49 females, 2 males) between the ages of 12 and 47 (means = 22.5) who met the criteria for AN of the Diagnostic and Statistical Manual of Mental Disorders, Third Edition, were randomly selected from an inpatient eating disorders unit during 15 months. Fifty healthy volunteers (41 females, 9 males) between the ages of 17 and 50 (means = 26.5) served as controls. After a neurologic history, all patients were evaluated by physical examination and standardized electrodiagnostic testing. Chi-square contingency testing was used to assess data. Four study group patients (8%) had electrodiagnostic evidence of a sensorimotor PN compared with none in the control group. This is approaching statistical significance (p = 0.13). Three of four patients with AN for at least ten years were among those with PN. The prevalence of subjective symptoms among the study group (65%) as compared to the control group (4%) was of marked significance (p = 5.62 x 10(-10]. In addition, three anorexic patients were found to have an isolated peroneal nerve palsy. We conclude that PN is a notable complication of AN, particularly in long-standing cases. The PN is most likely a product of chronic malnutrition rather than a specific nutrient deficiency. Patients with AN also appear to be at increased risk for developing localized compression neuropathies secondary to subcutaneous tissue loss. PMID- 2554848 TI - Delayed formation of gallstone after transcatheter arterial embolization for hepatocellular carcinoma. Is elective cholecystectomy advisable during hepatectomy? AB - Two hundred twenty-five transcatheter hepatic arterial embolizations (TAEs) were performed on 137 patients with hepatocellular carcinoma (HCC) during a three-year period. The postembolization changes of the gallbladder were studied by regular follow-up with ultrasonography. Twenty-four patients (10.7%) were found to have an acute infarction of the gallbladder within two weeks immediately following chemoembolization. Gallbladder infarction was related to inadequate superselectivity, regurgitation of chemoembolus, or unavoidable anatomic limitations. Four of the 24 patients were found to have delayed formation of gallstones, with the time lapses after TAEs being 2, 2, 3, and 5 months, respectively. One patient underwent surgical reexploration for cholecystectomy 14 months after resection of her HCC because of intractable symptoms and signs of chronic cholecystitis. There are four possible mechanisms of development of gallstones. Cholecystectomy should be performed during the elective hepatectomy for resectable HCC in patients who have received preoperative TAEs. PMID- 2554849 TI - Breast masses in young women. AB - To better define the risk of breast cancer in young patients, a retrospective review of all breast biopsies in women under age 40 years at Grady Memorial Hospital, Atlanta, from Dec 1, 1981, to Aug 15, 1987, was performed. During this time, 751 biopsies were performed on patients aged 9 to 40 years. None of the 128 patients aged 20 years or less had carcinoma. Of 150 patients aged 21 to 25 years, two had carcinoma. At age 26, there began a steady rise in the incidence of carcinoma, such that in the 36- to 40-year age group, carcinoma was present in 24.4% of the specimens. This retrospective review confirms previous reports that suggest that carcinoma of the breast is distinctly unusual in patients under age 20 and that breast masses in these young patients should be managed conservatively. As the incidence of carcinoma increases with the age of the patient, one's threshold for excisional biopsy should decrease. PMID- 2554850 TI - Effects of vitamin D derivatives on soft tissue calcification in neonatal and calcium mobilization in adult rats. AB - The activity of 18 vitamin D analogs on soft tissue calcification and growth impairment in neonatal rats and their effect on bone calcium mobilization, intestinal calcium absorption and binding to intestinal 1,25-dihydroxyvitamin D3 receptors in adult rats were compared. Depending on the chemical modification of the vitamin D parent compounds, they could be separated into active and inactive analogs. Cholecalciferol and ergocalciferol were similarly active, but epimerization of ergocalciferol at carbon 23 caused loss of activity. Hexafluorination at carbon 26 and 27 and the introduction of a double bond at carbon 22 or 23 had no or little effect on the activity. The loss of activity was caused by the introduction of a triple bond at carbon 23 and by hydroxylation at carbon 23, 26 or 28. The differentiation of human promyelocytic leukemia cells (HL-60) induced by these derivatives was used as a parameter for antitumour activity. All six analogs, which markedly affected calcium metabolism, were highly active in HL-60 cells. However, at least three derivatives were highly active in the antitumour test but failed to induce hypercalcemia. Thus, these results indicate that it could be possible to develop medically useful vitamin D derivatives devoid of hypercalcemic side-effects. PMID- 2554851 TI - Inactivation of end-plate acetylcholinesterase during the course of organophosphate intoxications. AB - Blood organophosphate (OP) levels, serum butyrylcholinesterase (BChE) activity and electrophysiological neuromuscular transmission following repetitive nerve stimulation at 10 Hz and 50 Hz were studied serially in five patients with severe acute organophosphate intoxication following suicide attempts. Eight to 45 hours after oral ingestion, blood OP levels were elevated, BChE activity was markedly reduced, while repetitive nerve stimulation studies showed no or only mild abnormalities. The latter attained the maximal abnormality 32-69 h after ingestion, when BChE was inactivated further but elevated OP levels had fallen. Recovery from these abnormalities at 10 Hz nerve stimulation occurred within 100 237 h after the intoxication and it was still incomplete at 50 Hz stimulation 48 80 h later. BChE activity varied within a wide range and showed even normal values at both times. Neuromuscular transmission studies proved to be the most useful indicator for determining the severity and time course of organophosphate intoxication. PMID- 2554852 TI - Duration of challenge immunity to coronavirus JHM in mice. AB - The duration of challenge resistance in mice immunized with mouse hepatitis virus (MHV) strain JHM was examined as a model of immunity to corona-virus infection. Genetically susceptible BALB/cByJ mice were immunized by intranasal (i.n.) or per os (p.o.) inoculation with MHV-JHM or sterile tissue culture fluid (sham) then challenged i.n. with MHV-JHM or sterile tissue culture fluid 1, 6, or 12 months later. Four days after challenge, virus in nasal turbinates and liver was quantified, and prevalence of microscopic lesions in liver and gut-associated lymphoid tissue was tabulated as indices of challenge resistance. MHV-immunized and challenged groups were compared to sham-immunized and challenged groups. Mice immunized by i.n. inoculation were strongly resistant to challenge at 1, 6, and 12 months. Mice immunized by p.o. inoculation were resistant at 1 month, but became partially susceptible to reinfection at 6 and 12 months, based upon all indices. These data indicate that, depending upon route of immunization, mice can become susceptible to reinfection with the same coronavirus strain over time. PMID- 2554853 TI - Langerhans cell density and activity in mouse skin and lymph nodes affect herpes simplex type 1 (HSV-1) pathogenicity. AB - Langerhans cells are epidermal antigen-presenting cells that function by taking up antigens in the skin, migrating to the lymph nodes, where they are designated interdigitating cells, and triggering the immune response. The role of interdigitating cells (IDC) was investigated in a murine model of herpes simplex virus-1 infection in the skin. The number of IDC in the lymph nodes began to increase on the first day following infection and reached a peak three days p.i. Low titers of infectious virus were recovered from the fraction of lymph node cells that consisted of 60-80% IDC at one day p.i. Lymph node cells that were obtained from mice immunized with HSV-1 proliferated in vitro in response to viral antigens but did not respond to mock antigens. When mice were immunized with HSV-1 inoculated into skin that had been depleted of Langerhans cells, this in vitro proliferative response was abolished. Thus, the present results suggest that Langerhans cells function in the immune defense of the skin against HSV-1 infection by transporting the virus to the peripheral lymph nodes where an immune response is initiated. Injection of the immunomodulator OK-432 into the footpad skin caused a local increase in the number of Langerhans cells in the epidermis and led to an increased migration of dendritic cells to the lymph nodes. Under these conditions, a decrease in HSV-1 pathogenicity was noted. These observations indicate that the pathogenicity of herpes simplex virus type 1 in the skin is affected by Langerhans cell density and activity in the epidermis and the lymph nodes. PMID- 2554854 TI - Rotavirus serotypes causing acute diarrhoea in hospitalized children in Yogyakarta, Indonesia during 1978-1979. AB - Rotavirus strains in stool specimens from 111 children aged 3-24 months admitted to hospital in Yogyakarta, Indonesia for treatment of acute diarrhoea were serotyped using VP7 serotype specific monoclonal antibodies in a double sandwich enzyme immunoassay. A serotype could be assigned to 59 of 111 specimens (53%). Inability to assign a serotype to 47% of specimens was probably due to loss of the outer capsid during transport of specimens from Indonesia to Australia. All four major human rotavirus serotypes were detected during the 15 month survey from June 1978 to August 1979, including one serotype 1, 5 serotype 2, 31 serotype 3, and 21 serotype 4 strains. One additional strain reacted with serotype 3 and 4 Mabs. Serotype 3 strains showed intratypic variation. The relative frequency of serotypes 2, 3, and 4 varied during the 15 months and appeared to be influenced by climatic changes associated with dry and wet seasons. Vaccine strategies must take account of comparatively rapid changes of predominant serotypes in a community and are only likely to be successful if comprehensive immunity can be established simultaneously against the four major human serotypes. PMID- 2554855 TI - Analysis of DNA synthesis in herpes simplex virus infected cells by dual parameter flow cytometry. AB - The DNA incorporation of a thymidine analogue bromodeoxyuridine (BrdUrd) in herpes simplex virus type 2 (HSV-2) infected cervical cancer cell lines (CaSki and C-33 A) was studied by dual parameter flow cytometry. HSV-2 infection resulted in an exponential increase in DNA synthesis in the CaSki cells. In the less permissive C-33 A cells the proportion of DNA-synthesizing cells cycled during HSV-2 infection. The inhibition of viral DNA synthesis by phosphonoformate (PFA) was able to inhibit the virus induced changes in the CaSki but not in the C 33 A cells. In the C-33 A cells a part of the virus induced cellular DNA synthesis represents repair replication of cellular DNA. PMID- 2554856 TI - Evaluation of continuous cell lines in antiviral studies with murine cytomegalovirus. AB - Cell culture systems were developed for rapid antiviral drug screening, using murine cytomegalovirus (MCMV) as an alternative to the slower growing human CMV. Since previous assay methods with MCMV employed mouse embryo fibroblasts (MEF cells), which are labor intensive to prepare and die off after 3-4 passages from primary culture, identification of virus-susceptible continuous cell lines was desirable. Three cell lines were found useful for assaying MCMV: C127I, SC-1, and 3T3. The antiviral agents acyclovir, ganciclovir, 5 fluoroarabinofuranosylcytosine, and 2'-fluoro-2'-deoxy-5 iodoarabinofuranosylcytosine were evaluated in the 3 continuous cell lines and in MEF cells. The 50% virus- or cell-inhibitory concentration values determined for each compound did not vary much from cell to cell. MEF cells were 10-fold more sensitive than the other cell lines to quantify virus from mouse organs, however. Virus propagated in 3T3 and SC-1 cells were as virulent to mice as salivary gland virus, whereas virus from MEF and C127I cells was more attenuated. Overall, C127I cells were judged to be the best for large scale antiviral screening in vitro, but MEF was the cell type of choice for titration of viruses from mouse organs and tissues. PMID- 2554857 TI - Inhibition of avian retrovirus protein synthesis in the presence of host cellular mRNA. AB - Two conditions of in vitro translation are described in which the RNA of Rous sarcoma virus is significantly less efficient at protein synthesis than host cellular chick mRNA. In direct competition, addition of increasing amounts of host cell mRNA inhibits synthesis of gag gene products from RSV RNA. When the denaturant dimethyl sulfoxide is added to a mixture of the two RNA's the inhibition is even greater. These results support the idea that protein synthesis from avian retrovirus RNA is negatively regulated in vivo in the presence of the host cell mRNA. PMID- 2554858 TI - Inhibition of SV40 DNA replication by Rous sarcoma virus LTR enhancer. AB - Simian virus 40 (SV40) late region recombinant constructs containing the Rous sarcoma virus (RSV) src gene along with RSV enhancer stimulated expression but completely abolished SV40 DNA replication. Constructs, in which the heterologous enhancer sequences were omitted, did replicate normally in African green monkey kidney cells and, in the presence of helper virus, gave rise to infectious progeny. Inhibition of SV40 DNA replication follows a cis-acting mechanism and is most likely due to a conformational change of the SV40 chromatin structure. PMID- 2554859 TI - A strategy simplifying site-directed mutagenesis in the CD4-binding region of HIV gp 120. AB - By site-directed mutagenesis, two unique restriction sites, surrounding the CD4 binding DNA region of HIV gp 120, were created. The mutations do not cause amino acid substitutions but generate a DNA fragment, which is stable when cloned into M 13 for further mutagenesis and is also easy to reclone. PMID- 2554861 TI - [Development of ACTH-containing cells in the hypothalamus and hypophysis in their normal state and in protein deficiency]. AB - In cryostate sections of the brain and hypophysis of newborn, 10- and 21-day-old rats, by means of the indirect method immunohistochemical reaction has been performed. As a primary serum rabbit antiserum against ACTH has been used. The ventromedial hypothalamus and adenohypophysis have been studied by means of light and electron microscopy. Relationship of ACTH-containing cells in the adenohypophysis in the process of the postnatal ontogenesis and protein insufficiency remains constant. Unlike the adenohypophysis, in the hypothalamus part of the ACTH-containing cells decreases with age. When the animals develop under conditions of protein insufficiency, changes in the part of the ACTH positive cells, that are age-dependent, do not coincide with corresponding parameters in the control. In the adenohypophyseal cells the products of diaminobensidine reaction are distributed in cytoplasm of granules with a dense center 100-200 nm in the diameter. The ACTH-containing cells in the hypothalamus are presented as small neurons with an even distribution of the immunohistochemical reaction products in cytoplasm and in large granules 200-280 nm in the diameter. Since in the adenohypophysis and in the hypothalamus various dynamics on contents of ACTH-reactive cells in the postnatal ontogenesis and at protein insufficiency has been revealed, and various localization of the diaminobenzidine reaction products has been demonstrated, a conclusion is made on heterogenous nature of antigenic determinants against ACTH, situating in these structures. PMID- 2554860 TI - Characterization of rotavirus subgroup-specific monoclonal antibodies and use in single-sandwich ELISA systems for rapid subgrouping of human strains. AB - Two subgroup-specific monoclonal antibodies (MAb) raised in mice against group A human rotavirus were shown to react by immunoblotting with the trimeric form of VP6 of the homologous subgroup and successfully applied to development of new single-sandwich ELISA systems for rapid subgrouping of human strains. All of the 344 strains tested could be subgrouped, but for two of them prior propagation in cell cultures was required. PMID- 2554862 TI - [Flow cytometry of DNA in research on kidney tumors]. AB - The DNA content was studied in 22 patients (including 10 children) with renal tumours. Renal clear cell carcinomas were mainly diploid (5 of 6 cases); one clear cell and other variants of renal cell carcinoma were aneuploid. There was a correlation between the degree of tumour cell anaplasia and ploidy of renal cell carcinomas: all diploid carcinomas were of I and II degree of anaplasia, aneuploid carcinomas, except one case, were of degree III of anaplasia. 5 out of 9 nephroblastomas were diploid, 4, aneuploid. The distinctive features of nephroblastoma were pronounced proliferative activity of tumour cells and a low DNA index of the aneuploid cell line which in all cases was localized in the vicinity of the diploid region. The remaining tumours (papillary epithelial nephroma, juxtaglomerular cell tumour and malignant schwannoma) were diploid with a relatively low proliferative activity of tumour cells. PMID- 2554863 TI - [Benign fibrous histiocytoma of bone]. PMID- 2554864 TI - Cultured cells as a screen for novel treatments of Alzheimer's disease. AB - Two easily measured abnormal properties of fibroblasts from patients with Alzheimer's disease have been utilized to develop an in vitro test system for screening novel therapeutic agents for Alzheimer's disease. The abnormal properties selected for study were increased isoproterenol-stimulated cyclic adenosine monophosphate production and decreased pH (measured by the weak-acid distribution method). L-Carnitine was tested as a potential therapeutic agent, since it has been used to treat a variety of experimental metabolic encephalopathies. The addition of L-carnitine normalized both of these properties in the Alzheimer cells. Tissue culture may aid as a preliminary screen for identifying novel approaches to the treatment of Alzheimer's disease. PMID- 2554865 TI - Human papillomavirus DNA in a recurrent squamous carcinoma of the eyelid. AB - The polymerase chain reaction (PCR) is a molecular technique that amplifies specific target DNA sequences in vitro to facilitate identification of DNA. We have applied the PCR to a recurrent infiltrating, well-differentiated squamous carcinoma from the right lower eyelid of a 37-year-old woman. With primers specific for human papillomavirus (HPV) type 16 DNA, PCR yielded a single band of amplified DNA product. The product was positive, with a radiolabeled HPV type 16 probe on dot blot analysis. The presence of HPV type 16 viral DNA in this recurrent squamous carcinoma of the eyelid has implications with regard to the possible origin, treatment, and prognosis of the tumor. PMID- 2554867 TI - A fluorescence test in allantoic cells for the detection of infectious bronchitis virus. PMID- 2554866 TI - Assessment of diclofenac on herpes keratitis in rabbit eyes. AB - Topical diclofenac sodium is a non-steroidal anti-inflammatory drug that is being developed for use in the control of postoperative and medical inflammation. A comparison was made of 0.1% diclofenac with 1% prednisolone sodium phosphate, 0.03% flurbiprofen, and a vehicle placebo in rabbit eyes with acute herpetic keratitis in a double-masked study. Maximum corneal epithelial involvement was observed in each group on day 6 postinoculation, and in eyes subsequently treated with prednisolone, the corneal epithelial involvement appeared to be more severe and to resolve more slowly. Conjunctivitis and corneal clouding peaked on days 6 to 7 for all treatment groups and remained most severe in the placebo-treated eyes, followed closely by those treated with prednisolone. The duration of virus shedding was the same for placebo-, flurbiprofen-, and diclofenac-treated groups (50% or more were virus negative by day 10 or 11). Only prednisolone-treated eyes had an extended period of virus shedding, and the rabbit mortality rate in this group was slightly higher. It thus appears that topical diclofenac does not exacerbate acute herpes keratitis; diclofenac-treated eyes displayed less or at least no more severe disease than did the eyes treated with the other anti inflammatory agents tested, and shedding of virus into tears was not prolonged. PMID- 2554868 TI - Reduced systolic blood pressure elevations during maximum exercise at simulated altitudes. AB - Ten healthy female subjects performed maximum exercise on a bicycle in an altitude chamber during normoxia and hypobaric hypoxia simulating altitudes of 2,450, 3,700 and 4,600 m. The increases in systolic blood pressure responses were reduced with the degree of hypobaric hypoxia, whereas heart rate and diastolic pressure responses were unchanged. The increases in blood levels of aldosterone, plasma renin activity, adrenaline, noradrenaline, neuropeptide-Y and vasoactive intestinal peptide were similar at the different simulated altitudes. Angiotensin converting enzyme and vasoactive intestinal peptide levels were not affected by hypoxia or maximum exercise. The present results suggest that the decreases in systolic blood pressure responses during hypobaric hypoxia could not be explained by altered responses of the measured vasoactive substances from the renin angiotensin, gastrointestinal, and autonomic nervous systems. PMID- 2554869 TI - Characterization of SV40-transformed human cells by immunofluorescence and fluorescent in situ hybridization techniques. AB - We have performed immunofluorescent and fluorescent in situ hybridization studies in order to better clarify the integration of SV40 DNA in human fibroblast cell lines. Most of the cells were T-antigen positive by immunocytochemical studies, while in all the cells we detected the integrated viral DNA by in situ hybridization. Both techniques are easy and useful to perform but the molecular genetic method gives a more specific signal with the possibility of localizing molecular hybrids in the nucleus and in the cytoplasm of the transformed cells. PMID- 2554870 TI - Positive inotropic stimulation in the normal and insufficient human myocardium. AB - Cardiac alpha- and beta-adrenoceptors and the positive inotropic effects of several adenylate cyclase dependent and independent agents have been measured in papillary muscle strips from patients without, as well as with moderate and severe heart failure. The number of beta-adrenoceptors was found to be decreased depending on the degree of heart failure. This does not apply to alpha adrenoceptors, which remain unchanged. The antagonist affinity of adrenoceptors for the different ligands did not change in heart failure. Maximal increases in force of contraction were measured after raising Ca++ up to 15 mM in the muscle strips. In healthy human myocardium, isoprenaline, dobutamine, IBMX or cardiac glycosides increase force of contraction to the same maximal values as Ca++ does. However, in cardiac tissue from heart failure patients, positive inotropic agents which increase intracellular cAMP or are cAMP-dependent are less effective than Ca++. Furthermore, the results seem to indicate a homologous (agonist specific) downregulation of receptors in moderate heart failure and a heterologous downregulation in severe heart failure. Thus, many well known positive inotropic drugs lose their effectiveness just when they are needed most: in severe heart failure. PMID- 2554872 TI - Studies on the cellular mechanisms of action of positive and negative inotropic agents. AB - We have reviewed the mechanism by which drugs that elevate cyclic AMP level modify myocardial contractility. We have presented preliminary evidence about the mechanism by which muscarinic agonists antagonize the effects of these drugs. Finally, we suggest that the protein phosphorylation experiments, particularly if done in dispersed myocytes, could be an efficient and cost-effective method of screening drugs which may act by elevating intracellular levels of cyclic AMP. PMID- 2554871 TI - Do human cardiac beta-2 adrenoceptors play a (patho)physiological role in regulation of heart rate and/or contractility? AB - There can be no doubt that in human heart in addition to beta 1-adrenoceptors, functional beta 2-adrenoceptors exist. Their (patho)physiological role in regulating heart rate and/or contractility, however, is still an open question. Under normal physiological conditions cardiac beta 2-adrenoceptors may not be of functional importance, since heart rate and contractility seem to be under the control of noradrenaline that in the human heart acts nearly exclusively at beta 1-adrenoceptors. However, in situations of stress when large amounts of adrenaline are released from the adrenal medulla additional stimulation of cardiac beta 2-adrenoceptors may contribute to increases in heart rate and/or contractility. Moreover, in endstage congestive cardiomyopathy where cardiac beta 1-adrenoceptors are selectively down-regulated, cardiac beta 2-adrenoceptors may substitute for the loss of beta 1-adrenoceptors to maintain (at least partially) contractility; under these conditions beta 2-adrenoceptor agonists, like dopexamine, may be of beneficial therapeutic effect. While a decrease in cardiac beta-adrenoceptor function appears to be a general phenomenon in all kinds of heart failure, it is not always due to a selective reduction in cardiac beta 1 adrenoceptors: in mitral valve disease both cardiac beta 1- and beta 2 adrenoceptors decline concomitantly in relation to the degree of heart failure. It is, therefore, doubtful whether under these conditions beta 2-adrenoceptor agonists may also be useful to support the failing heart. PMID- 2554873 TI - Mechanisms of positive inotropic effects. AB - This review deals with the principal mechanisms which are known to play a role in positive inotropism: 1) The myoplasmic Ca2+ concentration may be increased by increases in cyclic AMP. Beside receptor-mediated stimulation (isoprenaline) or direct stimulation (forskolin) of the adenylate cyclase, the cyclic AMP may be increased by phosphodiesterase inhibition; 2) Cyclic AMP-independent activation of Ca2+ channels can be brought about by alpha-adrenergic agents (phenylephrine) or so-called calcium agonists; 3) Only a small increase in myoplasmic Na+ concentration can greatly enhance the force of contraction by an increase in the intracellular Ca2+ concentration. This is possible by inhibition of the Na+/K+ ATPase (glycosides) or by prolongation of the open state of Na+ channels (DPI 201 106); 4) A direct inhibition of the Na+/Ca2+ exchange has been discussed for amiloride; 5) A prolongation of the action potential induced by K+ channel inhibiting agents such as 4-amino-pyridine may increase the myoplasmic Ca2+ concentration by a prolongation of the slow Ca2+ inward current; 6) An increased Ca2+ sensitivity of the contractile proteins has been demonstrated for a number of compounds in vitro; the contribution of such an effect to the overall positive inotropism is unknown because a calcium sensitizer without any effects on calcium or sodium movements is not yet available. PMID- 2554874 TI - Regulation of force and intracellular calcium transients by cyclic AMP generated by forskolin, MDL 17,043 and isoprenaline, and its modulation by muscarinic receptor agents: a novel mechanism for accentuated antagonism. AB - The relation of changes in intracellular calcium transients and force of isometric contractions in response to an elevation or reduction of cyclic AMP levels was investigated in isolated dog ventricular trabeculae and rabbit papillary muscles, in which multiple superficial cells have been microinjected with the calcium sensitive bioluminescent protein aequorin. Forskolin, MDL 17,043 and isoprenaline elevated the tissue cyclic AMP level, increased consistently the peak aequorin signals and force, and abbreviated the duration of both signals in a concentration-dependent manner. When the effect of isoprenaline was compared with that of alteration of extracellular calcium concentration [( Ca2+]0), the increase in force by isoprenaline was associated with higher peak aequorin signals than that by alteration of [Ca2+]0 for a given increase in force, indicating the decrease in calcium sensitivity of myofibrils by cyclic AMP generated by beta-adrenoceptor stimulation. Carbachol, which did not affect significantly the basal force and cyclic AMP levels, lowered the cyclic AMP levels elevated previously by forskolin, MDL 17,043 or isoprenaline in the isolated dog ventricular trabeculae. It antagonized the increase in peak aequorin signals and force caused by these agents in a concentration-dependent manner. When carbachol had been administered prior to isoprenaline and the concentration response curve for isoprenaline was determined in the presence of carbachol, the relation of force peak aequorin signals was not modified by carbachol in the rabbit papillary muscle. Carbachol, when administered during induction of the positive inotropic action by forskolin, MDL 17,043 and isoprenaline, decreased the force more than peak aequorin signals in a concentration-dependent manner in the dog ventricular trabeculae. Therefore, the relation of force to peak aequorin signals was shifted downwards during the carbachol-induced inhibition, indicating a further decrease of calcium sensitivity of myofibrils by carbachol. This effect of carbachol appears to be specific to the cyclic AMP-mediated positive inotropic action, since the alpha-adrenoceptor-mediated (cyclic AMP-independent) action was unaffected by carbachol. This mechanism may play an important role for "accentuated antagonism" in the mammalian ventricular myocardium. PMID- 2554875 TI - Studies on the mechanism of action of the bipyridine milrinone on the heart. AB - Milrinone is a positive inotropic and vasodilator agent when tested in experimental animals and in human heart-failure patients. It is generally believed that milrinone acts by inhibiting phosphodiesterase IV, thus increasing cyclic AMP, [Ca++]i and cardiac contractile force and relaxation. Maximal force produced by milrinone is greater when single-dose response curves are compared to cumulative dose-response curves. In vitro, milrinone produces a tachyphylaxis, the extent of which is both dose- and time-dependent. Recovery of tachyphylaxis is both dose- and time-dependent and is not influenced by inhibitors of protein or RNA synthesis. There is a specific cross-tachyphylaxis between milrinone and amrinone, theophylline, papaverine, and Bay K8644. This tachyphylaxis may explain the low maximal contractile response of the cumulative dose-response observed in isolated tissues. Milrinone increased cyclic AMP in dog and guinea pig cardiac muscle. As previously shown by Endoh et al., milrinone in low doses produced a biphasic effect on cyclic AMP. The early increase (first 60-70 s) in cyclic AMP shows a good correlation with contractile force changes. If cyclic AMP is determined at maximal contractile force this correlation was poor. Here we also present instances where the increase in cyclic AMP after milrinone (determined at maximal effect) does not correlate with the contractile response. The cross tachyphylaxis of milrinone with Bay K8644 suggests that milrinone has an action on the sarcolemmal Ca++ channels. Bay K8644 suppresses the positive inotropic effect of catecholamines by 50%, but not the cyclic AMP response. The inotropic effect of milrinone, in contrast to norepinephrine is highly sensitive to [Ca++]0, stimulation rate, and [K+]0. In this respect milrinone behaves more like Bay K8644. We postulate that the main inotropic action of milrinone is due to a sarcolemmal effect. The early cyclic AMP production described could be in the sarcolemmal compartment and this may explain some of the similarities of milrinone's actions with those of Bay K8644. The tachyphylaxis observed with the inotropic effect of milrinone does not extend to the decreases in relaxation time. This and other findings to be discussed suggest that the positive inotropic and reduction in relaxation time by milrinone depend on different mechanisms, possibly through differential compartmentalization of cyclic AMP. PMID- 2554877 TI - [Corpora amylacea of the brain and their significance for forensic medicine]. AB - In this study 66 section--cases have been examined in order to verify a possible coherence between the frequency of corpora amylacea (CA) of the brain and the age respectively the duration of mortal agony. The study was confined to the hippocampus, which is a zone of predilection for CA. The exploration with the light microscope showed a coherence between the frequency of CA and the biological age, but not with the duration of mortal agony. PMID- 2554876 TI - Adrenoceptors in myocardial regulation: concomitant contribution from both alpha- and beta-adrenoceptor stimulation to the inotropic response. AB - The studies presented deal with the alpha 1-adrenoceptor mediated inotropic effects of noradrenaline obtained by exclusive ("pure") alpha 1-adrenoceptor stimulation or by concomitant stimulation of alpha 1- and beta-adrenoceptors in myocardium. The pure beta-adrenergic effects of noradrenaline were also quantified. Interactions between the two receptor systems were studied. The pure alpha 1- and beta-adrenergic effects of noradrenaline, respectively, were achieved separately in the presence of high concentrations of appropriate receptor blockers. The experiments were performed on isolated ventricular myocardium from rat, rabbit, and man. The pure alpha 1-adrenergic inotropic effects were about 35-50% of control (basal) and half the pure beta-adrenergic effects both in rat and rabbit myocardium. Ventricular myocardium from man exhibited an alpha 1-adrenergic inotropic effect of the same magnitude (50% of control [basal]) as did rabbit papillary muscle. Determination of the alpha 1 adrenergic inotropic component during concomitant beta-adrenoceptor stimulation was associated with difficulties. Several experimental approaches on rat and rabbit myocardium are presented and discussed. Some types of experimental approaches obviously underestimate the alpha 1-adrenergic component. The methods regarded as reliable revealed an alpha 1-adrenergic inotropic effect of about 20 30% during combined adrenoceptor stimulation by noradrenaline. Concomitant beta stimulation reduced the alpha 1-adrenergic effect by about 50%, while alpha 1 stimulation attenuated the beta-effect to a lesser degree (about 20-25%). A model is presented on a mutual attenuation of the functional expression of the two receptor systems.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554878 TI - [Asbestos screening of autopsy material]. AB - This study was designed to provide basic data on the deposition of asbestos in human tissue in an urban area (Hamburg) with increased immission rates of fibres. The analysis of the unselected autopsy cases was carried out by light microscopy counting the typical asbestos bodies (method: wet digestion of tissue and filter technique). - RESULTS: a) The average concentration of asbestos bodies (AB) per ccm lung tissue (n = 80) amounts to 35 AB/ccm (men 38 AB/ccm, females 15 AB/ccm). b) Asbestos bodies could be found regularly in extrapulmonary tissues (n = 20, up to 13 different tissue samples per case), most frequently in lymph nodes, thyroid gland and spleen. c) Lungs of children in the age of 7 to 16 years (n = 40) showed asbestos bodies in 35% of the examined cases (with a range from 3 to 31 AB/ccm lung tissue). PMID- 2554879 TI - [THC-carbonic acid determination in serum with fluorescence polarization immunoassay (FPIA, TDX) and GCMS]. AB - The Urine-Immuno-Assay (Abbot, TDX) for Cannabis you may also use for the investigation of blood or serum. First the protein is precipitated by Metanol. The solvent is removed by a gentle stream of nitrogen and the solution reconstituted with Phosphat-Buffer. You may handle this solution like urine. Confirmation analysis is made by GCMS after extraction using C-18-extraction tubes and derivation with a proper reagent. Detection limit is about 10 ng/ml. You need only 0.3 ml Serum. PMID- 2554880 TI - [Comparative studies of the detection of rotavirus in fecal samples of calves with diarrhea with the latex test "Slidex Rota-Kit 2" and electron microscopy]. AB - Of 68 fecal samples from calves with diarrhoea which were tested for rotavirus with the latex agglutination test "Slidex Rota-Kit2" and by electron microscopy 33 samples were positive and 33 were negative with both tests respectively. Divergent results (latex test positive/EM negative and vice versa) were observed in one specimen only, respectively. Cross reactions with other viruses diagnosed by electron microscopy were not observed with the latex agglutination test. The "Slidex Rota-Kit2" is another suitable test for the diagnostic laboratory as well as for the veterinary practitioner for the detection of rotavirus in fecal samples of calves. PMID- 2554881 TI - Novel activity of angiotensin-converting enzyme. Hydrolysis of cholecystokinin and gastrin analogues with release of the amidated C-terminal dipeptide. AB - ACE (angiotensin-converting enzyme; peptidyl dipeptidase A; EC 3.4.15.1), cleaves C-terminal dipeptides from active peptides containing a free C-terminus. We investigated the hydrolysis of cholecystokinin-8 [CCK-8; Asp-Tyr(SO3H)-Met-Gly Trp-Met-Asp-Phe-NH2] and of various gastrin analogues by purified rabbit lung ACE. Although these peptides are amidated at their C-terminal end, they were metabolized by ACE to several peptide fragments. These fragments were analysed by h.p.l.c., isolated and identified by comparison with synthetic fragments, and by amino acid analysis. The initial and major site of hydrolysis was the penultimate peptide bond, which generated a major product, the C-terminal amidated dipeptide Asp-Phe-NH2. As a secondary cleavage, ACE subsequently released di- or tri peptides from the C-terminal end of the remaining N-terminal fragments. The cleavage of CCK-8 and gastrin analogues was inhibited by ACE inhibitors (Captopril and EDTA), but not by other enzyme inhibitors (phosphoramidon, thiorphan, bestatin etc.). Hydrolysis of [Leu15]gastrin-(14-17)-peptide [Boc (t butoxycarbonyl)-Trp-Leu-Asp-Phe-NH2] in the presence of ACE was found to be dependent on the chloride-ion concentration. Km values for the hydrolysis of CCK 8, [Leu15]gastrin-(11-17)-peptide and Boc-[Leu15]gastrin-(14-17)-peptide at an NaCl concentration of 300 mM were respectively 115, 420 and 3280 microM, and the catalytic constants were about 33, 115 and 885 min-1. The kcat/Km for the reactions at 37 degrees C was approx. 0.28 microM-1.min-1, which is approx. 35 times less than that reported for the cleavage of angiotensin I. These results suggest that ACE might be involved in the metabolism in vivo of CCK and gastrin short fragments. PMID- 2554882 TI - Desensitization of calcium mobilization and cell function in human neutrophils. AB - Neutrophils pretreated with the chemoattractant formylmethionyl-leucyl phenylalanine become unresponsive when re-exposed to the same ligand, a process termed desensitization. We have examined whether desensitization of transduction (Ca2+ mobilization) or of other cell functions (superoxide generation, enzyme release, or aggregation) occurs synchronously. Simultaneous studies of Ca2+ mobilization and aggregation by using Fura-2-loaded cells indicate that, under conditions where the aggregation response is abolished, most of the Ca2+ mobilization is unaltered. Further studies were then carried out to ascertain whether desensitization of Ca2+ mobilization could in fact be induced. Desensitization was observed, and was dependent on the number of exposures of the cells to the ligand, the concentration of the ligand, and whether the ligand was left in the medium or was removed. The pattern of resensitization was dependent on the experimental design. Under conditions where ligand was continuously present, no recovery of the Ca2+-mobilization response was seen with subsequent challenges. In contrast, on removal of ligand, this response showed partial recovery. Whereas complete desensitization of aggregation was noted, enzyme release showed a markedly lesser degree of desensitization and required more frequent exposures to the ligand before it was observed. Little or no desensitization of superoxide generation was observed regardless of the conditions utilized. Studies using phorbol myristate acetate as the ligand showed that Ca2+ mobilization and aggregation could be simultaneously inhibited. Our results suggest that discrete mechanisms of desensitization are possible in human neutrophils, and that desensitization of one particular function (aggregation) does not imply concomitant desensitization of other functions. PMID- 2554883 TI - Inhibition of IMP-specific cytosolic 5'-nucleotidase and adenosine formation in rat polymorphonuclear leucocytes by 5'-deoxy-5'-isobutylthio derivatives of adenosine and inosine. AB - 1. The partially purified IMP-specific cytosolic 5'-nucleotidases from rat liver, polymorphonuclear leucocytes and heart were inhibited by 50% by 2-6 mM-5'-deoxy 5'-isobutylthioadenosine (IBTA) or 7-10 mM-5'-deoxy-5'-isobutylthioinosine (IBTI). IBTA and IBTI inhibited the rat liver and polymorphonuclear-leucocyte enzymes non-competitively. IBTA, but not IBTI, also inhibited the ecto-5' nucleotidase of polymorphonuclear leucocytes. IBTI was, by contrast, a more potent inhibitor than IBTA of the AMP-specific soluble 5'-nucleotidase from pigeon heart. 2. During 2-deoxyglucose-induced ATP-catabolism in rat polymorphonuclear leucocytes, adenosine formation was inhibited by approx. 80% by 3 mM-IBTA and by approx. 70% by 7 mM-IBTI. 3. The results show that 5'-modified nucleosides are inhibitors of cytosolic 5'-nucleotidases and that they penetrate to inhibit their target enzymes in intact cells. Such inhibitors may be useful to clarify the mechanisms of adenosine formation and to prevent mononucleotide hydrolysis during ATP breakdown. PMID- 2554884 TI - The role of cytochrome c4 in bacterial respiration. Cellular location and selective removal from membranes. AB - The cellular location of cytochrome c4 in Pseudomonas stutzeri and Azotobacter vinelandii was investigated by the production of spheroplasts. Soluble cytochrome c4 was found to be located in the periplasm in both organisms. The remaining cytochrome c4 was membrane-bound. The orientation of this membrane-bound cytochrome c4 fraction was investigated by proteolysis of the cytochrome on intact spheroplasts. In P. stutzeri, 78% of the membrane-bound cytochrome c4 could be proteolysed, whilst 82% of the spheroplasts remained intact, suggesting that the membrane-bound cytochrome c4 is on the periplasmic face of the membrane in this organism. Cytochrome c4 was not susceptible to proteolysis on A. vinelandii spheroplasts, in spite of being digestible in the purified state. Cytochrome c5 was shown to have a similar cellular distribution to cytochrome c4. Selective removal of cytochrome c4 from membranes of P. stutzeri was accomplished by the use of sodium iodide and propan-2-ol, with the retention of most of the ascorbate-TMPD (NNN'N'-tetramethylbenzene-1,4-diamine) oxidase activity associated with the membrane. Sodium iodide removed most of the cytochrome c4 from A. vinelandii membranes with retention of 62% of the ascorbate-TMPD oxidase activity. Cytochrome c4 could be returned to the washed membranes, but with no recovery of this enzyme activity. We conclude that cytochrome c4 is not involved in the ascorbate-TMPD oxidase activity associated with the membranes of these two organisms. PMID- 2554885 TI - Studies on some specific Ap4A-degrading enzymes with the use of various methylene analogues of P1P4-bis-(5',5'''-adenosyl) tetraphosphate. AB - Six new methylenephosphonate analogues of P1P4-bis-(5',5'''-adenosyl) tetraphosphate, Ap4A, having P2-P3 carbon bridges CF2, CCl2 and CH2CH2 or P1-P2 and P3-P4 carbon bridges CF2, CCl2 and CH2CH2 in the tetraphosphate chain, were examined as substrates or inhibitors for two specific Ap4A-degrading enzymes: (asymmetrical) Ap4A hydrolase (EC 3.6.1.17) from yellow-lupin seeds and (symmetrical) Ap4A hydrolase (EC 3.6.1.41) from Escherichia coli. All analogues in which the central oxygen atom was replaced by a stable carbon bridge were hydrolysed by the asymmetrical hydrolase (CF2 greater than CCl2 greater than O greater than CHBr greater than CH2 greater than CH2CH2). As expected, these analogues were not hydrolysed by the symmetrical hydrolase, which was also unable to act on analogues having P1-P2 and P3-P4 carbon bridges. PMID- 2554886 TI - Desensitization of alpha 2-adrenergic receptors in NG 108 15 cells by (-) adrenaline and phorbol 12-myristate 13-acetate. AB - alpha 2-Adrenergic receptors on NG 108 15 cell membranes were identified by [3H]rauwolscine binding: Bmax. = 661 +/- 81 fmol/mg of protein, Kd = 6.9 +/- 2.5 nM (mean +/- S.E.M., n = 6). On intact cells, stimulation of these receptors by ( )-adrenaline inhibited the prostaglandin-E1-stimulated adenylate cyclase activity by about 60%. The effect of (-)-adrenaline was pertussis-toxin-sensitive, indicating the involvement of an inhibitory G protein. (-) Adrenaline/[3H]rauwolscine competition-binding experiments revealed that only 50% of the alpha 2 receptors were coupled to G proteins (i.e. displayed high agonist affinity). Pre-treatment of the cells with 20 microM-(-)-adrenaline provoked homologous desensitization of the alpha 2 receptors. The alpha 2-adrenergic response decreased after a time lag of about 2 h, to reach a minimum after 12 h. The bradykinin and muscarinic responses were not affected. The alpha 2-receptor concentration decreased without time lag. The high-agonist-affinity sites disappeared more rapidly (t1/2 = 42 min) than did the low-affinity uncoupled sites (t1/2 approx. 20 h). In contrast, pertussis-toxin-mediated [32P]ADP ribosylation of inhibitory G proteins was unaffected by the pre-treatment. Pretreatment of intact NG 108 15 cells with 1 microM-phorbol 12-myristate 13 acetate (PMA) provoked a rapid decrease of the alpha 2-adrenergic response. The effect was nearly complete after 40 min. PMA also decreased the bradykinin response, suggesting a heterologous desensitization process. The alpha 2-receptor concentration, the (-)-adrenaline competition-binding curves and the pertussis- and cholera-toxin-mediated [32P]ADP-ribosylation of their respective G proteins were not affected. PMID- 2554887 TI - Bordetella pertussis adenylate cyclase inactivation by the host cell. AB - Bordetella pertussis produces a calmodulin-dependent adenylate cyclase (AC) which acts as a toxin capable of penetrating eukaryotic cells and generating high levels of intracellular cyclic AMP. Transfer of target cells into B. pertussis AC free medium leads to a rapid decay in the intracellular AC activity, implying that the invasive enzyme is unstable in the host cytoplasm. We report here that treatment of human lymphocytes with a glycolysis inhibitor and an uncoupler of oxidative phosphorylation completely blocked the intracellular inactivation of B. pertussis AC. Lymphocyte lysates inactivated all forms of B. pertussis AC in the presence of exogenous ATP. This inactivation was associated with degradation of an 125I-labelled 200 kDa form of B. pertussis AC. It appears that ATP is required for the proteolytic pathway, but not as an energy source, since non-hydrolysable ATP analogues supported inactivation and complete degradation of the enzyme. The possibility that binding of ATP to B. pertussis AC renders it susceptible to degradation by the host cell protease is discussed. PMID- 2554888 TI - Potassium channels in the luminal membrane of rabbit proximal straight tubule. Evidence from vesicle studies. AB - The characteristics of 86Rb+ fluxes through K+ channels in luminal-membrane vesicles isolated from the pars recta of rabbit proximal tubule were studied. In KCl-loaded vesicles from the pars recta, transient accumulation of 86Rb+ is observed which is modestly inhibited by BaCl2 and blocked by CdCl2. The isotope accumulation is driven by an electrical diffusion potential, as shown in experiments using either these membrane vesicles loaded with different anions, or an outwardly directed Li+ gradient with a Li+ ionophore. The vesicles containing the channel show a cation selectivity with the order K+ greater than Rb+ greater than choline+ greater than or equal to Li+ greater than Na+. The CdCl2-sensitive 86Rb+ flux is dependent on intravesicular Ca2+. Increasing concentrations of Ca2+ gradually decreased the 86Rb+ uptake and at 1 microM-Ca2+ the CdCl2-sensitive isotope flux is nearly abolished. PMID- 2554889 TI - Differential effects of leupeptin, monensin and colchicine on ligand degradation mediated by the two asialoglycoprotein receptor pathways in isolated rat hepatocytes. AB - We have shown that degradation of asialo-orosomucoid (ASOR) in isolated rat hepatocytes occurs by two different intracellular pathways [Clarke, Oka & Weigel (1987) J. Biol. Chem. 262, 17384-17392] mediated by two subpopulations of cell surface galactosyl (Gal) receptors, designated State 1 or State 2 receptors. In the present study, several inhibitors were tested for their effects on ligand degradation by the State 1 or State 2 pathway. Leupeptin, monensin and chloroquine completely inhibited degradation of 125I-labelled ASOR in both pathways. Dose-response studies showed, however, that the State 2 pathway was more sensitive to leupeptin or monensin than the State 1 pathway. No differences were observed with chloroquine. For example, the onset of inhibition in the State 2 and State 1 pathways occurred at about 0.05 and 0.3 microM-leupeptin respectively, a 6-fold difference. At 3.5 microM-monensin, 125I-ASOR degradation in the State 2 pathway was completely blocked, whereas degradation in the State 1 pathway was essentially unaffected. Colchicine was observed to give the largest differential sensitivity between the two pathways. The State 2 degradation pathway was about 30-fold more sensitive to colchicine than the State 1 pathway. Lumicolchicine had no affect. The onset of inhibition of the rate of 125I-ASOR degradation in the State 2 and State 1 pathways occurred at approximately 0.1 and 3.0 microM-colchicine respectively. At very high concentrations (greater than 0.1 mM), the State 1 pathway could be completely inhibited. We conclude that intracellular 125I-ASOR processing or delivery to degradative compartments in both the State 1 and State 2 Gal receptor pathways requires low pH. Ligand delivery to the degradative compartment does not require microtubules in the State 1 pathway, consistent with the very rapid onset of degradation in this pathway. The State 2 degradation pathway does require microtubules. PMID- 2554890 TI - Down-regulation of protein kinase C potentiates angiotensin II-stimulated polyphosphoinositide hydrolysis in vascular smooth-muscle cells. AB - In smooth-muscle cells (SMC) isolated from rat aorta, angiotensin II stimulates a phospholipase C with subsequent formation of inositol trisphosphate (InsP3). Short-term (10 min) pretreatment of SMC with 12-O-tetradecanoylphorbol 13-acetate (TPA; 100 nM) decreases the angiotensin II-induced InsP3 formation. However, this inhibition is not observed after incubating the cells for 2 h with TPA. Longer term pretreatments even lead to an enhanced generation of InsP3. This increased response to angiotensin II occurs without a significant change in the receptor number or Kd value of angiotensin II binding to the cells. The biologically inactive phorbol ester 4 alpha-phorbol 12,13-didecanoate was without effect on angiotensin II-stimulated InsP3 generation, irrespective of the time of preincubation. In parallel with this potentiation of angiotensin II-induced generation of InsP3 by TPA, a down-regulation of protein kinase C activity is observed. A 24 h pretreatment of SMC with TPA decreases protein kinase C activity to less than 10% of that of control cells. Longer-term pretreatment also increases the angiotensin II-induced release of Ca2+ and delays the decay of the transient Ca2+ increase. All these data suggest that protein kinase C exerts a negative feedback control on angiotensin II-stimulated polyphosphoinositide turnover, and that protein kinase C is an important factor in limiting the production of InsP3 in stimulated cells. PMID- 2554891 TI - 5'-Nucleotidases of chicken gizzard and human pancreatic adenocarcinoma cells are anchored to the plasma membrane via a phosphatidylinositol-glycan. AB - We have analysed the membrane anchorage of plasma-membrane 5'-nucleotidase, an ectoenzyme which can mediate binding to components of the extracellular matrix. We demonstrated that the purified enzyme obtained from chicken gizzard and a human pancreatic adenocarcinoma cell line were both completely transformed into a hydrophilic form by treatment with phospholipases C and D, cleaving glycosylphosphatidylinositol (GPI). These data indicate the presence of a glycolipid linker employed for membrane anchoring of the 5'-nucleotidase obtained from both sources. Incubation of plasma membranes under identical conditions revealed that about half of the AMPase activity was resistant to GPI-hydrolysing phospholipases. Investigation of the enzymic properties of purified chicken gizzard 5'-nucleotidase revealed only minor changes after removal of the phosphatidylinositol linker. However, cleavage of the membrane anchor resulted in an increased sensitivity towards inhibition by concanavalin A. After tissue fractionation, chicken gizzard 5'-nucleotidase could be obtained as either a membrane-bound or a soluble protein; the latter is suspected to be released from the plasma membrane by endogenous phospholipases. Higher-molecular-mass proteins immuno-cross-reactive with the purified chicken gizzard 5'-nucleotidase were detected as both soluble and membrane-bound forms. PMID- 2554892 TI - Identification of high levels of type 1 and type 2A protein phosphatases in higher plants. AB - Extracts of Brassica napus (oilseed rape) seeds contain type 1 and type 2A protein phosphatases whose properties are indistinguishable from the corresponding enzymes in mammalian tissues. The type 1 activity dephosphorylated the beta-subunit of phosphorylase kinase selectively and was inhibited by the same concentrations of okadaic acid [IC50 (concentration causing 50% inhibition) approximately 10 nM], mammalian inhibitor 1 (IC50 = 0.6 nM) and mammalian inhibitor 2 (IC50 = 2.0 nM) as the rabbit muscle type 1 phosphatase. The plant type 2A activity dephosphorylated the alpha-subunit of phosphorylase kinase preferentially, was exquisitely sensitive to okadaic acid (IC50 approximately 0.1 nM), and was unaffected by inhibitors 1 and 2. As in mammalian tissues, a substantial proportion of plant type 1 phosphatase activity (40%) was particulate, whereas plant type 2A phosphatase was cytosolic. The specific activities of the plant type 1 and type 2A phosphatases were as high as in mammalian tissue extracts, but no type 2B or type 2C phosphatase activity was detected. The results demonstrate that the improved procedure for identifying and quantifying protein phosphatases in animal cells is applicable to higher plants, and suggests that okadaic acid may provide a new method for identifying plant enzymes that are regulated by reversible phosphorylation. PMID- 2554894 TI - Properties of the superoxide-generating oxidase of B-lymphocyte cell lines. Determination of Michaelis parameters. AB - The capacity of three B-lymphocyte cell lines to generate superoxide (O2.-) was examined. The Burkitt lymphoma lines P.3HR-1 and Jijoye gave no response to phorbol 12-myristate 13-acetate (PMA) at 100 ng/ml but produced up to 0.35 nmol of O2.-/min per mg of protein when stimulated with 5 micrograms of PMA/ml; the cell line RPMI 1788 produced Nitro Blue Tetrazolium-positive responses to low PMA concentrations and approx. 0.4 nmol of O2.-/min per mg of protein at 5 micrograms of PMA/ml. Each cell line contained approx. 10 pmol of low-potential cytochrome b (cytochrome b-245)/mg of protein. Homogenates of PMA-activated cells gave 10-20 fold greater rates of O2.- produced per mg of protein. The Km for NADPH varied between approx. 250 microM for P3.HR-1 and RPMI 1788 cell lines and 30.5 +/- 6.5 microM for the Jijoye cell line; the Km values for NADH were higher. Determination of intracellular NADPH concentration showed that this might limit the rate of O2.- production since in each cell line it was at or below the Km concentration. PMID- 2554893 TI - E.p.r.-spectroscopic studies on the molybdenum-iron site of nitrogenase from Clostridium pasteurianum. AB - The e.p.r. spectroscopy of the nitrogenase molybdenum-iron protein from Clostridium pasteurianum was re-investigated. The sharpness of the delta Ms = +/- 3 g'z peak from the +/- 3/2 Kramer's doublet enables the observation and quantification of incompletely resolved hyperfine splittings from the stable magnetic nuclei 95Mo and 57Fe in samples enriched in these isotopes. No couplings to 1H or 17O could be discerned by examination of spectra from samples exchanged into 2H2O and H2(17)O respectively. Simulation of the spectrum from 95Mo-enriched samples yields a hyperfine coupling of 2.9 MHz, and indicates that the earlier electron-nuclear-double-resonance-derived estimate of 8.1 +/- 0.2 MHz is substantially in error. PMID- 2554896 TI - Inactivation of liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase by o phthalaldehyde. AB - The two activities of chicken liver 6-phosphofructo-2-kinase/fructose-2,6 bisphosphatase were inactivated by o-phthalaldehyde. Absorbance and fluorescence spectra of the modified enzyme were consistent with the formation of an isoindole derivative (1 mol/mol of enzyme subunit). The inactivation of 6-phosphofructo-2 kinase by o-phthalaldehyde was faster than the inactivation of fructose-2,6 bisphosphatase, which was concomitant with the increase in fluorescence. The substrates of 6-phosphofructo-2-kinase did not protect the kinase against inactivation, whereas fructose-2,6-bisphosphate fully protected against o phthalaldehyde-induced inactivation of the bisphosphatase. Addition of dithiothreitol prevented both the increase in fluorescence and the inactivation of fructose-2,6-bisphosphatase, but not that of 6-phosphofructo-2-kinase. It is proposed that o-phthalaldehyde forms two different inhibitory adducts: a non fluorescent adduct in the kinase domain and a fluorescent isoindole derivative in the bisphosphatase domain. A lysine and a cysteine residue could be involved in fructose-2,6-bisphosphate binding in the bisphosphatase domain of the protein. PMID- 2554897 TI - Inhibitory action of atrial natriuretic factor on cholinergic and nonadrenergic, noncholinergic neurotransmission in guinea pig small intestine. AB - We studied the effect of synthetic rat atrial natriuretic factor (ANF) (Ser 99 Tyr 126) on the isolated guinea pig proximal ileum. This preparation contained about one-third of the endogenous tissue ANF content which, for the most part, comes from the blood. ANF inhibited, in a dose-dependent manner, cholinergic twitch contractions (EC50 = 4.2 nM), nonadrenergic, noncholinergic (NANC) primary and rebound contractions and histamine-induced sustained tonic contraction (but not carbachol induced contraction) of the longitudinal muscle. Ascending enteric reflex (AER) contractions of the circular muscle were inhibited though not dose dependently. We suggest pre- and post-synaptic actions of sustained intestinal tissue and blood ANF levels which may play a role in regulating motor activity and muscle tone of the small intestine. PMID- 2554895 TI - Heterogeneity of smooth endoplasmic reticulum from rat liver studied by two-phase partitioning. AB - Smooth microsomal membranes, prepared from rat liver by sucrose-density-gradient centrifugation, were subfractionated by counter-current distribution in an aqueous two-phase system consisting of poly(ethylene glycol) and Dextran T500. A comparison of the distribution curves of marker enzymes, together with theoretically calculated curves, indicated the presence of at least five membrane subfractions, differing in the ratios of the marker enzymes. Glucose-6 phosphatase and arylesterase distributed in one manner, and NADPH-cytochrome c reductase and NADH-ferricyanide reductase in another. Evidence for further heterogeneities in the distribution of marker enzymes in smooth microsomes was obtained by analysing the membrane domain structure using a recently described method [Albertsson (1988) Q. Rev. Biophys. 21, 61-98]. Phenobarbital treatment did not influence the behaviour of the marker enzymes. PMID- 2554898 TI - Pineal muscarinic phosphoinositide response: pertussis toxin resistant signaling with very low receptor number. AB - Autoradiography revealed very low density of muscarinic receptors in the rat pineal gland. Yet, the magnitude of phosphoinositide hydrolysis elicited with 0.1 mM carbachol was similar to that seen with 1 mM norepinephrine. The cholinergic and adrenergic phosphoinositide responses were fully additive. The cholinergic signal was insensitive to pertussis toxin both in vivo and in vitro and persisted in pineals cultured for 5 days. The data expand our previous finding on functional muscarinic acetylcholine receptors in the rat pineal gland. PMID- 2554899 TI - Computer-assisted imaging cytometry of nuclear chromatin reveals bone tumor virus infection and neoplastic transformation of adherent osteoblast-like cells. AB - Established osteoblast-like (OB) cells infected with the bone tumor-inducing C type retrovirus OA MuLV remained nontumorigenic over 104 cell culture passages. DNA histograms revealed a new cell population with a stem line peak at 5c. A second OA MuLV-infected OB cell line underwent neoplastic transformation with increasing passage level. These cells showed diffuse aneuploidy. Stepwise linear discriminant analysis of the chromatin structure of control, OA MuLV-infected, and FBR osteosarcoma virus-transformed cell lines resulted in various levels of discrimination ranging between 79.6% for control cells versus nontumorigenic OA MuLV-infected cells, and 96.6% for nontumorigenic OA MuLV-infected cells versus FBR osteosarcoma virus-transformed cells. OA MuLV-infected tumorigenic cells and FBR osteosarcoma virus-transformed cells were discriminated at a 93.6% level. PMID- 2554900 TI - A novel family of growth factor receptors: a common binding domain in the growth hormone, prolactin, erythropoietin and IL-6 receptors, and the p75 IL-2 receptor beta-chain. AB - Lymphokine and hematopoietic growth factors control the differentiation and proliferation of diverse cell types by binding to specific cell-surface receptors. Strikingly, the recently elucidated sequences of the interleukin-6 and erythropoietin receptors, and the interleukin-2 receptor beta-chain (p75), display a significant evolutionary resemblance of their extracellular domains. This homology extends to the binding domains of the growth hormone/prolactin class of receptors. Alternatively, little similarity exists between the cytoplasmic extensions of these diverse receptors. I discuss the evolutionary and functional implications of this broad, mosaic receptor relationship, with particular reference to possible structural resemblances between the cognate growth factors. PMID- 2554901 TI - Angiotensin II and phorbol-esters potently down-regulate endothelin (ET-1) binding sites in vascular smooth muscle cells. AB - [125I]ET-1 binding to vascular smooth muscle cells showed an apparent single class of high affinity recognition sites with a Kd of 2.12 +/- 0.46 nM and a Bmax of 81.2 +/- 5.2 fmol/10(6) cells. The specific binding was equally and totally displaced by ET-1 and ET-2 whereas ET-3 presented a different pattern. We investigated heterologous regulation of ET-1 binding sites by preincubating the cells with angiotensin II (AII), Arg-vasopressin, bradykinin, enkephalins, serotonin, norepinephrine and carbachol, for 18 h at 37 degrees C. Only AII pretreatment resulted in an important and dose-dependent decrease of ET-1 binding capacity. Sar1-Ile8-AII inhibited the regulatory effect of AII. Furthermore, preexposure of the cells with phorbol-12,13 dibutyrate but not with phorbol-12,13 didecanoate also resulted in a concentration-dependent diminution of ET-1 binding sites. These findings suggest that AII may selectively down-regulate ET-1 binding sites in vascular smooth muscle cells by a mechanism involving protein kinase C. PMID- 2554902 TI - Abolition of the NMDA-mediated responses by a specific glycine antagonist, 6,7 dichloroquinoxaline-2,3-dione (DCQX). AB - Among various quinoxaline derivatives examined, only 6,7-dichloroquinoxaline-2,3 dione (DCQX) competitively displaced the strychnine-insensitive binding of [3H]glycine, without affecting the other binding sites on the N-methyl-D aspartate (NMDA) receptor complex. This novel specific antagonist abolished the ability of L-glutamate to potentiate [3H]MK-801 binding activity in brain synaptic membranes treated with Triton X-100. Inclusion of glycine reversed this preventive action of DCQX on the potentiation induced by glutamate. PMID- 2554903 TI - Tiazofurin and selenazofurin induce depression of cGMP and phosphatidylinositol pathway in L1210 leukemia cells. AB - The synthetic nucleoside tiazofurin(2-beta-ribofuranosylthiazole-4-carboxyamide) and its selenium analog selenazofurin inhibited the growth of L1210 leukemia cell culture in a dose dependent manner with IC50 value of 2.0 and 0.2 Um respectively. The GTP/ATP ratio was diminished 4-6 fold as measured by HPLC, while IMP/ATP increased 6-8 fold. The decreased guanylate pools may explain the 30% reduction in cyclic GMP levels and GTPase activity measured after the treatment with the nucleosides. Inhibition of phospholipase C activity is suggested since diacylglycerol content, protein kinase C activity and phorbol ester binding of the membrane fraction were also reduced 20-40%. These results reveal a novel aspect in the action of these compounds which may play a role in their therapeutic action and selectivity. PMID- 2554904 TI - Calcium dependent cysteine proteinase is a precursor of a chemotactic factor for neutrophils. AB - A new chemotactic factor for neutrophils is generated from calcium dependent cysteine proteinase (calpain) I by autodigestion. An active peptide was isolated from the autodigest and its structure was determined to be an acetylated nonapeptide with the sequence: N-acetyl Ser-Glu-Glu-Ile-Ile-Thr-Pro-Val-Tyr. Compared with the entire sequence of human calpain I, the peptide was identical with the N-terminal amino acid sequence of the large subunit deduced from the cDNA sequence, except that the peptide was devoid of a methionine residue and acetylated at the N-terminus. The acetyl nonapeptide was synthesized and its chemotactic activity was reconfirmed. The biological significance and possible role of this calpain derived chemotactic factor in inflammation are discussed. PMID- 2554905 TI - Light-induced DNA cleavage by esperamicin and neocarzinostatin. AB - Ultraviolet radiation of the enediyne drugs is effective in causing nicks in supercoiled DNA. Of special interest is the fact that the observed nucleotide cleaving specificity for the UV light- and thiol-activated antibiotics was the same with esperamicin A1, but was different with neocarzinostatin. In addition to the preferred cutting of T and A bases, the light-activated neocarzinostatin attacked certain G bases which were rarely cleaved by the thiol-activated neocarzinostatin. It should be noted that these enediyne antibiotics lose the DNA breakage activity after light-exposure for 30 min. PMID- 2554906 TI - 2',3'-dideoxycytidine permeation of the human erythrocyte membrane. AB - The mechanism by which 2,3'-dideoxycytidine, an inhibitor of HIV-I infectivity, permeates the cell membrane was investigated. The influx of ddCyd into human erythrocytes was nonconcentrative. The initial velocity of both ddCyd influx and efflux was, in contrast to compounds that permeate the cell membrane via the nucleoside transporter, a linear function of nucleoside concentration in the 1 microM to 10 mM range and relatively insensitive to temperature. Furthermore, potent inhibitors of nucleoside transporter and other nucleosides were found to inhibit ddCyd influx only partially or not at all suggesting that ddCyd permeates the human erythrocyte membrane predominantly by nonfacilitated diffusion. This unusual characteristic seems to be due to the lack of 3'-hydroxyl moiety of ddCyd which appears to be an important determinant for the nucleoside carrier specificity rather than to lipid solubility itself. As far as permeation of the cell membrane is concerned ddCyd shares these properties with 2',3' dideoxythymidine and 3'-azido-3'-deoxythymidine. PMID- 2554907 TI - Coordinate increase in collagenase mRNA and enzyme levels in human fibroblasts treated with the tumor cell factor, TCSF. AB - Cocultures of human fibroblasts and LX-1 human lung carcinoma cells expressed 8 10 fold higher levels of collagenase mRNA than the sum of the individual cells, in parallel with similar increases in collagenase activity. Addition of the tumor cell collagenase stimulatory factor, TCSF, purified from LX-1 cells, to these fibroblasts also gave a 3-4 times increase in collagenase mRNA level. However, various fibroblast cell lines differed in their response to TCSF stimulation. Thus one cell line, GM 1391, did not respond to TCSF but responded to another potent collagenase stimulator, tetradecanoyl phorbol ester. Another cell line 5383 did not respond significantly to either agent. In each case collagenase activity and mRNA levels responded in a parallel fashion. PMID- 2554908 TI - Differential susceptibility to biological detergents of the particulate cGMP stimulated phosphodiesterase from rat heart: preservation of the allosteric properties of the solubilized enzyme. AB - The effects of various biological detergents on the particulate cGMP-stimulated cAMP phosphodiesterase activity from rat heart were investigated. When added to particulate fractions, anionic and non-ionic detergents diversely increased both cAMP and cGMP phosphodiesterase activities and slightly decreased the stimulatory effect of cGMP on cAMP hydrolysis whereas cationic detergents were rather inhibitory and drastically lowered the stimulatory effect of cGMP. Among the most efficient detergents, only sodium cholate was able to solubilize phosphodiesterase activity and preserve the stimulatory effect of cGMP on cAMP hydrolysis. Furthermore, the addition of glycerol significantly improved the conservation of the allosteric properties of the enzyme. Kinetic properties of the cholate-solubilized phosphodiesterase were quite identical to those of the membrane-bound enzyme. PMID- 2554910 TI - Generation of free radicals by alloxan in the presence of bovine serum albumin: a role of protein sulfhydryl groups in alloxan cytotoxicity. AB - The interaction of alloxan with bovine serum albumin was studied. When alloxan was incubated with bovine serum albumin, oxygen consumption, H2O2 formation, and diminution of sulfhydryl groups of the protein were observed. During the reaction of alloxan with the protein, superoxide radicals were generated; and under anaerobic conditions, ESR signal of alloxan free radicals was observed. These results strongly suggest that alloxan mediates electron transfer from the protein sulfhydryl groups to oxygen. PMID- 2554909 TI - Demonstration of adenylate-kinase activity in hepatic microsomes. Relevance to Ca2+ uptake. AB - It is demonstrated that the hepatic microsomal fraction contains significant adenylate-kinase activity. This explains a paradoxical ADP-stimulated Ca2+ uptake into microsomal vesicles which is inhibited when adenylate kinase is inhibited. The presence of adenylate kinase in the microsomal fraction helps to prevent sudden drops in ATP level, and thus has a stabilizing effect on the many ATP dependent reactions carried out in this subcellular compartment. PMID- 2554911 TI - Inactivation of rat gastric mucosal histidine decarboxylase by phosphatase. AB - Histidine decarboxylase of supernatants as well as of purified preparations from rat gastric mucosa is inactivated by a non-specific phosphatase in the absence of pyridoxal 5'-phosphate. The inactivation is a time and concentration-dependent process. Pyridoxal 5'-phosphate, but not histidine, protects the enzyme against phosphatase action. The inactivation is reversible, only pyridoxal 5'-phosphate reactivates the inactivated enzyme. Pyridoxamine 5'-phosphate is ineffective for histidine decarboxylase, but is converted into an active coenzyme only in gastric supernatant. Evidence for the occurrence of an active phosphatase in gastric tissue is also presented; its properties are those of an acid phosphatase and are similar to those of phosphatases hydrolyzing pyridoxal 5'-phosphate in other tissues. The data indicate that phosphatase promotes apoenzyme formation and may play a role in the regulation of histamine synthesis. PMID- 2554912 TI - Calf thymus deoxyuridine triphosphatase differs from rat spleen enzyme in molecular disposition. AB - Antiserum against calf thymus dUTPase was raised in rats by injection of the partially purified enzyme. The antiserum did not react with dUTPase purified from rat spleen, while antibody against rat spleen dUTPase partially reacted with calf thymus enzyme. Native molecular weight of the calf thymus dUTPase was estimated at 46,000 daltons by gel filtration, and the denatured form of the enzyme was about 22,000, as judged by immunoblot analyses using the antibodies. These results indicate that the calf thymus dUTPase is composed of two identical subunits. PMID- 2554913 TI - In vitro effect of eicosapentaenoic and docosahexaenoic acids on prostaglandin E2 synthesis in a human lung carcinoma. AB - The capacity to synthesize both prostaglandins E1 (PGE1) and E2 (PGE2) has been determined in human lung mucoepidermoid carcinoma homogenates when [14C]-fatty acid precursors were added to the incubation medium. Only 10% of the total radioactivity recovered in PGs was found in PGF1 alpha and PGF2 alpha. The experiments were principally focused to inhibit the PGE2 synthesis either with pure eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids or with mixtures of both n-3 fatty acids obtained from fish oil. The results demonstrated that significant inhibitions were found when using 25 microM or a higher concentration of pure EPA or DHA in the incubation medium; however, 5 microM of mixtures of different EPA/DHA ratio caused the same inhibition. The results suggest that EPA and DHA, when added together, may enforce their inhibitory effect on PGE2 synthesis. PMID- 2554914 TI - Studies on the formation of lactate and pyruvate from glucose in cultured skin fibroblasts: implications for detection of respiratory chain defects. AB - We investigated the time course of the formation of lactate and pyruvate from glucose in cultured skin fibroblasts from controls, from a patient with a cytochrome c oxidase deficiency and from controls treated with inhibitors of the individual respiratory chain complexes. Fibroblasts from the patient and inhibitor treated fibroblasts produced more lactate and less pyruvate; this resulted in a significant increase in the lactate to pyruvate ratio, reflecting an increased cytosolic NADH/NAD+ redox state. We conclude that measurement of lactate and pyruvate production from glucose in cultured skin fibroblasts can be of value in the diagnosis of inherited diseases of the mitochondrial respiratory chain. PMID- 2554915 TI - Characterization of two possible forms of type IV collagen from human kidney cortex. AB - Type IV collagen was isolated from human kidney cortex according to the purification of mouse kidney type IV collagen (MKIVC) with minor modifications as described previously [Oikawa, T., et al. (1986) Chem. Pharm. Bull. 34, 789-797]. Chromatography of human kidney type IV collagen (HKIVC) on a column of DEAE cellulose resulted in its separation into two fractions, i.e., one (HKIVC-1) passed through and the other (HKIVC-2) adsorbed to the column, similar to MKIVC. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Bio-Gel A-5m chromatography revealed that both HKIVC-1 and -2 differed from each other in the ratios of major and minor components. Amino acid analyses also demonstrated that there was a significant difference in the contents of several amino acid residues between both type IV collagens and their 80-kDa components, one of the major ones. These results indicate the possibility that there exist two forms of type IV collagen in human kidney. PMID- 2554916 TI - Multiple forms of calpastatin in pig brain. AB - Pig brain was found to contain two calpain-specific, heat-stable inhibitory fractions which could be separated by DEAE-cellulose chromatography. CS-0.1, which was eluted from the column at 0.1 M NaCl, was identified as an ordinary, well-known calpastatin. CS-0.2, eluted at 0.2 M NaCl, was different from CS-0.1 in that it inhibited calpain 1 more strongly than calpain II and that it did not cross-react with anti-calpastatin antibodies. Partial purification indicated that CS-0.2 contained inhibitor proteins smaller than ordinary calpastatin, but whether they are the products derived from CS-0.1 or entirely different genetic products has not yet been determined. PMID- 2554917 TI - Identification of functional beta-adrenergic receptors on AC glioma cells. AB - AC glioma cells, a clonal cell line derived from a rat glioma, responded to 1 mM dibutyryl-cyclic AMP and isobutylmethylxanthine with a change to stellate morphology. A concentration-related morphological change was induced by beta 1- and beta 2-adrenergic agonists with the order of potency being isoproterenol greater than soterenol greater than norepinephrine. Propranolol (nonselective, beta-antagonist), butoxamine (beta 2-antagonist) and metoprolol (beta 1 antagonist) significantly decreased the cell response to isoproternol. Schild analysis of the response, using the competitive antagonist metoprolol, gave pA2 values of 7.5 and 8.5 for the agonists norepinephrine and soterenol, respectively, with slopes of the curves being less than unity. These observations indicate that both beta 1- and beta 2-adrenergic receptors mediate the change in cellular morphology. PMID- 2554918 TI - Synaptosomal (Ca2+-Mg2+)-ATPASE activity modulation by cyclic AMP. AB - Dibutyryl cyclic AMP, in a concentration-dependent manner, increased synaptosomal (Ca2+-Mg2+)-ATPase activity, but in synaptic plasma membranes lacked any effect. The maximal enzyme activity in synaptosomes was increased by 38%, leaving unaltered the extrasynaptosomal Ca2+ concentration necessary to reach it. In the presence of 5 microM cyclic AMP, cyclic AMP-dependent protein kinase increased (30%) maximal (Ca2+-Mg2+)-ATPase activity in synaptic plasma membranes, but the apparent affinity for Ca2+ was not modified. This effect was partially inhibited (60%) by a cyclic AMP-dependent protein kinase inhibitor. The data suggest that synaptosomal (Ca2+-Mg2+)-ATPase activity is modulated by a cyclic AMP-dependent phosphorylation reaction. PMID- 2554919 TI - Effects of adenosine analogues on basal, prostaglandin E1- and forskolin stimulated cyclic AMP formation in intact neuroblastoma cells. AB - We have examined the effects of R-phenylisopropyladenosine (R-PIA) and other adenosine analogues on basal, prostaglandin E1 (PGE1)- and forskolin-stimulated cyclic AMP (cAMP) formation in intact N1E-115 neuroblastoma cells, to determine whether the cells contain A1 adenosine receptors that are negatively coupled with adenylate cyclase. Basal levels of cAMP (68 +/- 7 pmol/mg protein; mean +/- SE, N = 15) were not altered by low concentrations of R-PIA. The apparent lack of inhibition was not due to increases in cAMP due to activation of a stimulatory A2 receptor by endogenously-synthesized adenosine. By comparison, low levels of R PIA did reduce significantly (P less than 0.05) PGE1-dependent increases in cAMP formation (maximum response to PGE1, 972 +/- 77 pmol cAMP/mg protein; EC50 for PGE1, 0.2 microM). Inhibition was dose dependent, and resulted in a 30-50% maximum reduction in production stimulated by PGE1. Nanomolar concentrations of R PIA elicited half-maximal inhibition; the inhibitory response was blocked by 8 phenyltheophylline (8-PT). The order of potencies of several adenosine analogues in eliciting this response suggested that inhibition was mediated by an A1 adenosine receptor. Examination of the effects of R-PIA on forskolin-stimulated cAMP formation yielded several interesting findings. First, stimulation by the diterpene by itself was blocked by both adenosine deaminase (ADA) and 8-PT (40 and 25% inhibition respectively). Low concentrations of R-PIA (less than 10(-6) M) had no effect on forskolin-stimulated cAMP production. At higher levels (greater than or equal to 10(-6) M) the analogues acted synergistically with the diterpene, to yield cAMP levels that were up to 3-fold higher than the additive effect of the two agents. Potentiation was stereospecific, Ca2+ dependent, and was blocked by 8-PT. The results of this study suggest that, in N1E-115 neuroblastoma cells, inhibitory A1 receptors are not stimulated in response to non-specific elevations in cAMP, but are associated with specific stimulatory receptors such as those activated by PGE1. PMID- 2554920 TI - Regulation of adenylate cyclase by cannabinoid drugs. Insights based on thermodynamic studies. AB - The abilities of lipophilic cannabinoid drugs to regulate adenylate cyclase activity in neuroblastoma cell membranes were analyzed by thermodynamic studies. Arrhenius plots of hormone-stimulated adenylate cyclase activity exhibited a break point at 20 degrees. The break point was reduced to 14 degrees by benzyl alcohol, consistent with results from other laboratories that have correlated this response with the increase in membrane fluidity induced by benzyl alcohol. Because cannabinoid drugs partition into membrane lipids and alter membrane fluidity parameters in a number of model systems, it was of interest to examine the influence of delta 9-tetrahydrocannabinol and cannabidiol on enzyme activity analyzed by the Arrhenius plot. delta 9-Tetrahydrocannabinol, known to inhibit adenylate cyclase, failed to decrease the transition temperature either at 1 microM or at concentrations exceeding its aqueous solubility (30 microM), suggesting that delta 9-tetrahydrocannabinol could not mimic the effects observed with benzyl alcohol. In contrast, 30 microM cannabidiol, which stimulated enzyme activity slightly, decreased the Arrhenius plot break point to 17.5 degrees. The decrease in the transition temperature in response to benzyl alcohol or cannabidiol was not accompanied by a change in activation energies above or below the transition temperature. delta 9-Tetrahydrocannabinol inhibits adenylate cyclase activity via Gi as does the muscarinic agonist carbachol (Howlett et al., Mol Pharmacol 29: 307-313, 1986). Both carbachol and delta 9-tetrahydrocannabinol decreased the enthalpy and entropy of activation. The net free energy of activation at 37 degrees was increased in the presence of both of these inhibitory agonists. These data suggest that, for the entropy-driven hormone stimulated adenylate cyclase enzyme, less disorder of the system occurs in the presence of regulators that inhibit the enzyme via Gi. In summary, thermodynamic data suggest that cannabidiol can influence adenylate cyclase by increasing membrane fluidity, but that the inhibition of adenylate cyclase by delta 9 tetrahydrocannabinol is not related to membrane fluidization. PMID- 2554921 TI - Effects of selective inhibitors on cyclic nucleotide phosphodiesterases of rabbit aorta. AB - In this study three forms of cyclic nucleotide phosphodiesterase (PDE) isolated from rabbit aorta were pharmacologically characterized, and the consequence of selective inhibition of calmodulin-stimulated PDE (CaM-PDE) and cGMP specific PDE (cG-PDE) was evaluated using PDE inhibitors. The cG-PDE (F1) was selectively inhibited by M&B 22948 (IC50 = 0.5 microM) and dipyridamole (IC50 = 7 microM). The cAMP-PDE (cA-PDE, F3) was inhibited more effectively by the cA-PDE inhibitor milrinone than by other PDE inhibitors. The cA-PDE preparation appeared to contain both cG-inhibited PDE and cG-insensitive PDE based on an additive inhibition of the activity by milrinone and SQ 65442, respective inhibitors of these enzymes. Vinpocetine, 8-methoxymethyl isobutylmethylxanthine (8-MeOMeMIX) and M&B 22948 effectively inhibited CaM-PDE (F2). Vinpocetine was a more selective inhibitor of CaM-PDE than M&B 22948 or 8-MeOMeMIX. CaM-PDEs isolated from rabbit aorta and bovine brain exhibited a similar sensitivity to these inhibitors. Seventy-two percent of the cGMP-hydrolyzing activity of this rabbit aortic CaM-PDE preparation was immunoadsorbed to monoclonal antibody (ACC-1) against CaM bound to brain CaM-PDE. Vinpocetine, 8-MeOMeMIX and M&B 22948 at concentrations (30 and 100 microM) which inhibit CaM-PDE greater than 60% increased cGMP but not cAMP levels in l-norepinephrine (NE) preincubated rabbit aortic slices. At concentrations selectively inhibiting cG-PDE, dipyridamole and M&B 22948 increased cGMP levels in untreated slices but failed to increase cGMP levels significantly in NE-treated slices. By contrast, vinpocetine failed to increase cGMP significantly in untreated slices, although it increased cGMP levels in NE or KCl preincubated slices. These data indicate that, in activated (precontracted) aorta, CaM-PDE is a major enzyme, whereas in untreated aorta cG PDE is a predominant enzyme for the hydrolysis of cGMP. This study also shows a usefulness of selective inhibitors in identifying different forms of PDE and similar drug sensitivities and immunoadsorption of aortic and brain CaM-PDEs by a monoclonal antibody. PMID- 2554922 TI - Role of thromboxanes in alterations of the diabetic beta-adrenergic system. AB - The inotropic effects of isoproterenol (ISO), as well as the beta-adrenoceptors population, were measured in cardiac tissues from normal and short-term (3 days) diabetic rats. ISO increased the tension of both normal and diabetic ventricles, but the efficacy (Emax) of the concentration-response curve was greater on ventricles from diabetic rats than in those from the normal control. This phenomenon was accompanied by a decrease in the number of beta-adrenoceptor sites (Bmax) during diabetes. Insulin-treated diabetic hearts partially reversed the phenomenon. Propanolol blocked, in a competitive manner, the positive inotropic action of ISO in both types of ventricles. Inhibition of the synthesis and receptors of thromboxane (TX) reduced the hyperreactivity to ISO and increased the number of beta-adrenoceptors during diabetes, producing Bmax values almost similar to those of the normal heart. Additionally, the diabetic heart generated and released a greater amount of TXB2 than the normal heart, even in the presence or absence of ISO. The stimulatory effect of ISO upon TXB2 release was altered by the specific beta-adrenergic blockade and by verapamil. In addition, the drugs able to induce a sustained increase of endogenous cAMP also inhibited the release of TXB2 by diabetic ventricles. Exogenous TXB2 exerted the same type of hyperreactivity in diabetic ventricles. This phenomenon was accompanied by an inhibition of Na+ + K+-ATPase activity. These results suggest that beta adrenergic inotropic stimulation is secondary to receptor-mediated hydrolysis of arachidonic acid with subsequent release of thromboxanes, which, in turn, may be responsible for both the superreactivity and the decrease in the number of beta adrenoceptors during diabetes. The abnormal reactivity to beta-agonists also could be associated with alterations of the diabetic cardiac Na+ + K+-ATPase activity induced by TXB2 whose production is increased during diabetes. PMID- 2554923 TI - Neurotensin(8-13): comparison of novel analogs for stimulation of cyclic GMP formation in neuroblastoma clone N1E-115 and receptor binding to human brain and intact N1E-115 cells. AB - Neurotensin(8-13), the carboxyl-terminal portion of neurotensin, is 4-50 times more potent than native neurotensin in binding to intact neuroblastoma N1E-115 cells and human brain tissue and in stimulation of intracellular cyclic GMP production and inositol phospholipid hydrolysis in clone N1E-115 (Gilbert JA and Richelson E, Eur J Pharmacol 99: 245-246, 1984; Gilbert JA et al., Biochem Pharmacol 35: 391-397, 1986; Kanba KS et al., J Neurochem 46: 946-952, 1986; and Kanba KS and Richelson E, Biochem Pharmacol 36: 869-874, 1987). A series of novel analogs of neurotensin (8-13) was synthesized, and a structure-activity study was done comparing the abilities of these peptides to stimulate intracellular cyclic GMP production in intact neuroblastoma clone N1E-115 and to inhibit the binding of [3H]neurotensin to these cells and to membranal preparations from human brain. A direct correlation was found for each analog between its EC50 for biochemical activity and its KD for binding ability in studies with clone N1E-115. Furthermore, a strong correlation existed for each peptide between its KD for binding to neurotensin receptors on these cells and its KD for binding to neurotensin receptors in human brain tissue. In this study, the residues that were important to the biochemical and binding activities of neurotensin (8-13) proved to be identical to the amino acids that are necessary for the functional integrity of native neurotensin (Gilbert JA et al., Biochem Pharmacol 35: 391 397, 1986. PMID- 2554924 TI - Dideoxycytidine permeation and salvage by mouse leukemia cells and human erythrocytes. AB - Transmembrane equilibration of dideoxycytidine (ddCyd) in P388 mouse leukemia cells and human erythrocytes was only 1% as rapid as that of uridine and 2' deoxycytidine which is mediated by the facilitated nucleoside transporter of these cells. ddCyd entry was nonsaturable up to a concentration of 1 mM but was partially inhibited by dipyridamole, nitrobenzylthioinosine and nucleosides, but not by nucleobases. Thus, entry was partly (70-80%) mediated, though very inefficiently, by the nucleoside carrier. Intracellular phosphorylation of ddCyd in P388 cells was also very inefficient compared to that of 2'-deoxycytidine and uridine and not rate limited by its slow entry into the cells. PMID- 2554925 TI - Reversal of ethinylestradiol-induced cholestasis by epomediol in rat. The role of liver plasma-membrane fluidity. AB - Epomediol (EPO) is a synthetic terpenoid compound shown to be active in increasing bile flow and some enzymatic activities of liver plasma membranes in the rat. The possible effect of EPO treatment in the ethinyl-estradiol (EE) induced cholestasis in the rat was investigated by measuring the hepatic transport of sulfobromophthalein (BSP) (plasma clearance and biliary secretion) and bile flow. Liver plasma membrane fluidity was also determined by the steady state fluorescence polarization (P) of diphenylhexatriene (DPH). EE administration (5 mg/kg s.c. for 5 days) was followed by a significant, comparable reduction (P less than 0.001) in BSP plasma clearance and biliary excretion and in bile flow. Intraperitoneal administration of EPO (100 mg/kg) to EE-treated rats restored both parameters of BSP transport, as well as bile flow, to control values. Liver plasma membrane fluidity was markedly (P less than 0.01) decreased by EE administration with a concomitant reduction (P less than 0.01) in Na+/K+-ATPase activity. EPO administration significantly increased membrane fluidity to values higher either to cholestatic (P less than 0.05) or control (P less than 0.05) animals. On the contrary, EPO did not influence Na+/K+-ATPase activity in either EE-treated or control animals. These data indicate that EPO fully reverses the impairments of BSP transport and bile flow induced by EE, possibly by reversing the decrease in liver plasma membrane fluidity induced by the synthetic estrogen. On the contrary, the EE-mediated decrease in Na+/K+ ATPase activity was not reversed by EPO. PMID- 2554926 TI - Selective mouse breeding for short ethanol sleep time has led to high levels of hepatic aromatic hydrocarbon (Ah) receptor. AB - Following a selective breeding program of heterogeneous mice for more than 30 generations, SS ("short sleep") and LS ("long sleep") lines have been developed on the basis of their sleep times when challenged with a single intraperitoneal dose of ethanol. The aromatic hydrocarbon responsiveness (Ah) locus encodes the Ah receptor, which regulates the induction of certain drug-metabolizing enzymes by polycyclic aromatic compounds such as 3-methylcholanthrene and tetrachlorodibenzo-p-dioxin. The C57BL/6 inbred mouse strain (B6; Ahb/Ahb) has a high-affinity Ah receptor, while the DBA/2 inbred mouse strain (D2; Ahd/Ahd) has a low-affinity Ah receptor. We show here that the SS inbred mouse line exhibits markedly elevated hepatic levels of the high-affinity Ah receptor, while the LS outbred mouse line contains the low-affinity Ah receptor. Among progeny of (B6D2)F1 X D2 backcross, the b/d heterozygote (having the high-affinity Ah receptor) was found to be several times more resistant than the d/d homozygote to a single dose of intraperitoneal ethanol. The D2.B6-Ahb congenic line is also several times more resistant to intraperitoneal ethanol than the B6.D2-Ahb congenic line is also several times more resistant to intraperitoneal ethanol than B6.D2-Ahd congenic line. We found that the waking blood ethanol levels are the same in b/d and d/d mice, suggesting that the relative ethanol resistance in b/d mice cannot be explained on the basis of a difference in central nervous system sensitivity. There are no differences between SS and LS mice or between b/d and d/d mice with regard to (i) blood acetaldehyde levels after a single intraperitoneal dose of ethanol, or (ii) hepatic alcohol dehydrogenase activities. There is a difference in the rate of ethanol elimination: SS more rapid than LS; b/d more rapid than d/d. Although SS mice have lower hepatic aldehyde dehydrogenase activities (cytosolic, mitochondrial low-Km: and mitochondrial high-Km forms) than LS mice, b/d and d/d do not show this difference. These data suggest that a selected mouse breeding program, based on resistance to a single intraperitoneal dose of ethanol, selects concurrently for the hepatic high-affinity Ah receptor. This selective advantage cannot be explained on the basis of changes in alcohol dehydrogenase or aldehyde dehydrogenase activities and might provide insight into the nature of the endogenous ligand for the Ah receptor. PMID- 2554927 TI - In vitro modulation of human neutrophil superoxide anion generation by various calcium channel antagonists used in ischemia-reperfusion resuscitation. AB - Generation of toxic oxygen species by activated polymorphonuclear leukocytes (PMNs) may represent an important mechanism of ischemia-reperfusion injury. Concentration-response data concerning inhibition of superoxide anion (O2-) generation by NADPH oxidoreductase (NADPH OR) from isolated human PMN were generated for five calcium channel antagonists commonly utilized in ischemia reperfusion investigational therapeutics. Regression analysis derived IC50 values for verapamil, nimodipine, nicardipine and lidoflazine were 45, 20, 12 and 7 microM respectively. Inhibition of the extent of reaction at 5 min paralleled inhibition of initial velocity. No inhibition by flunarizine was noted at concentrations less than or equal to 25 microM (where it did not alter reaction mixture composition). Only nicardipine demonstrated a significant concentration response effect relative to prolonging lag time preceding O2- synthesis. Inhibition appeared at least partially reversible for all five agents. Neither PMN activation/desensitization, free-radical scavenging, nor PMN cytotoxicity appeared to be involved in the inhibition of PMN O2- synthesis by these agents. Ca2+ antagonist inhibition of PMN NADPH OR appears to involve more than simple inhibition of Ca2+ flux across the PMN plasma membrane. Direct inhibition of the intracellular events involved in the activation and/or activity of NADPH OR may be operative. PMID- 2554928 TI - Role of hepatic and renal cytochrome P-450 IVA1 in the metabolism of lipid substrates. AB - The role of clofibrate-inducible cytochrome P-450 IVA1 in the metabolism of endogenous lipids in both rat liver and kidney microsomal fractions has been investigated. 20(omega)-hydroxyarachidonic acid has been identified as a major metabolite after incubation with both tissue fractions and the structure confirmed by mass spectrometry. The arachidonic acid 20-hydroxylase activity is inducible by clofibrate in both liver and kidney, indicating that cytochrome P 450 IVA1 is probably the enzyme responsible for this activity. In addition, the kidney exhibited higher rates of arachidonate 20-hydroxylase activity than the liver (in both control and induced states). Although leukotriene B4 was also hydroxylated in the 20-position in both liver and kidney, clofibrate induction resulted in a decrease (approximately 50%) in hydroxylase activity. In addition, the absolute level of leukotriene B4 20-hydroxylase activity in both tissue homogenates and by purified cytochrome P-450 IVA1 in a reconstituted system, was 2-3 orders of magnitude lower than the corresponding activity for lauric acid and arachidonic acid as substrates, indicating that the leukotriene was not the preferred substrate for this enzyme. Computer modelling of the conformational geometries of the above three potential cytochrome P-450 IVA1 substrates have shown that both lauric and arachidonic acids adopt a compact, 'hairpin' structure that are almost superimposed on each other, thereby rationalizing why they are relatively good substrates for this isoenzyme. By contrast, leukotriene B4 adopts a more bulky geometry than the two fatty acids, thereby providing a coherent structural reason why it is a poorer substrate for the cytochrome P-450 IVA1 isoenzyme. PMID- 2554929 TI - Characteristic beta-adrenergic receptors in a rat ascites hepatoma cell line (AH130). PMID- 2554930 TI - [Reverse phase high performance liquid chromatography in the synthesis of leukotrienes A4 and C4]. AB - The chromatographic (RP HPLC) behaviour of leukotriene C4, its methyl ester, leukotriene A4 methyl ester and some chemicals involved in their synthesis have been investigated. Optimal conditions of separation were determined for the gradient and isocratic HPLC. Parameters of the interaction of the substances with hydrophobic surface are discussed in terms of solvophobic theory. PMID- 2554931 TI - Increased levels of proteoglycan fragments in knee joint fluid after injury. AB - We measured levels of cartilage proteoglycan (PG) fragments in knee joint synovial fluid obtained from patients with previous trauma of the knee, early gonarthrosis, or pyrophosphate synovitis, and in age-matched control subjects. During the initial 3-4 weeks after rupture of the anterior cruciate ligament or the meniscus (confirmed by arthroscopy), markedly increased PG fragment levels were found. At later times after trauma (up to 4 years), many of these patients still had significantly elevated levels of cartilage PG fragments in the joint fluid. In a group of older patients with gonarthrosis, these levels were only moderately elevated, while in patients with acute pseudogout, greatly increased levels were observed. Although longitudinal studies are needed to validate the significance, PG fragments in joint fluid may be a marker for early posttraumatic arthrosis. PMID- 2554932 TI - Inflammatory microcrystals stimulate interleukin-6 production and secretion by human monocytes and synoviocytes. AB - Crystal-related joint diseases are often associated with systemic inflammatory manifestations, including increased levels of acute-phase proteins, leukocytosis, and fever. Recently, interleukin-6 (IL-6) has been identified as a pluripotent mediator of inflammatory and immunologic responses and the major hepatocyte stimulating factor. In this study, we demonstrated that monosodium urate (MSU) and calcium pyrophosphate dihydrate (CPPD) crystals, and to a lesser extent, hydroxyapatite crystals, increased IL-6 production by synoviocytes and monocytes in vitro. Immunoprecipitation experiments showed that MSU and CPPD crystals, but not hydroxyapatite crystals, were able to increase the release of newly synthesized IL-6. Crystal-induced IL-6 stimulated acute-phase protein synthesis, immunoglobulin production, and hybridoma cell proliferation, which was neutralized by a specific antibody to IL-6. High levels of IL-6 were found in synovial fluid from patients with gout and pseudogout. These results demonstrate that MSU and CPPD crystals can induce IL-6 production in synoviocytes and monocytes, and that synovial fluid from patients with gout and pseudogout contains high levels of IL-6. Crystal-induced IL-6 is likely to be an important mediator of inflammatory responses in acute gout and pseudogout. PMID- 2554934 TI - Protein kinase C modulation of the ethanol effect on serotonin2 receptor transduction in astrocytes. AB - Acute ethanol exposure stimulated serotonin2 receptor signalling in cultured astrocytes. Pretreatment with the protein kinase C inhibitor H7 significantly increased the ethanol-induced potentiation of [3H]-inositol phosphates accumulation. The increase could be explained by an augmented activation of phospholipase C. The results indicate a role of PKC for the modulation of ethanol effects on cellular signalling. PMID- 2554933 TI - In vitro pharmacological properties of a series of isoprenaline derivatives. AB - The p-trifluoromethylanilide congener of isoprenaline, tert-butyl N-[(S)[( 4-[(R) 6-[2-(3,4-dihydroxyphenyl-2- hydroxyethyl]amino]heptanamido]phenyl]methyl][(N methylcarbamoy l) methyl]carbamoyl]methyl]carbamate (1S,4R)-4,7,7-trimethyl-3-oxo 2- oxabicyclo[2.2.1]heptane-1-carboxylate (1:1) (Ro 17-2218) was investigated for its effects in various pharmacological tests in vitro and compared to the parent compound. As Ro 17-2218 represented a mixture of four diastereomers, the pure isomers were synthesized. They had a purity of 97-98%. By pharmacological testing of the diastereomers the highest potency was found in the 6R,2'R-isomer Ro 17 8648, while the potency of the 6S,2'S-isomer, Ro 17-9651 was lower by three orders of magnitude. The amorphous hydrochloride Ro 17-8648/001 had 1/10 the potency of the respective crystalline camphanate Ro 17-8648/003. (R)-6-[(R)-[2 (3,4-Dihydroxyphenyl)-2-hydroxyethyl]amino]-N-[4- (trifluoromethyl) phenyl]heptan amide (Ro 17-8648/003) was found to have potent beta-agonist properties with clear beta 1-selectivity in radioligand binding studies. It exerted an extremely tight binding to membrane receptors. As a full beta-agonist it elicited positive inotropic effects in isolated cardiac tissues, with a potency 10-360 times that of isoprenaline and an extremely long duration of action. Electrophysiological studies in isolated guinea-pig papillary muscles confirmed the beta 1-receptor mediated effects of the compound. PMID- 2554935 TI - Disulfiram neuropathy: a review (1971-1988) and report of a case. AB - Neuropathy is one of the most severe side effects of disulfiram therapy. We report the case of a young man who developed a neuropathy following disulfiram administration, with a virtually complete recovery in 14 months. We then discuss 37 cases of disulfiram neuropathy reported since 1971. Evidence is given that: (1) there is no numerical sex prevalence, although the incidence of the disease in women is probably disproportionately high; (2) symptom onset latency is dose dependent, being longer at 250 mg/day or less; (3) neurological deficits are also dose-dependent, being milder at 250 mg/day or less; (4) the two previous findings and single observations suggest that disulfiram neuropathy is a dose-dependent phenomenon; (5) recovery probably follows a course which depends primarily on the initial degree of impairment; (6) the genetic mechanism probably involves carbon disulfide and a hypothesis as to the possible biochemical mechanism is proposed; (7) chloral hydrate can bear a potentiation effect on neuropathy, and the association with disulfiram is best avoided. Further, we give guidelines for the differentiation between alcoholic and disulfiram neuropathy, advise prescribing the drug at 250 mg daily or less, if possible, and stress the utmost importance of an early diagnosis. PMID- 2554936 TI - High fiber, low fat diets lead to lower cholesterol levels. PMID- 2554937 TI - [Schwannoma of the nose and paranasal sinuses]. AB - A rare case of a Schwannoma (neurinoma) of the frontal sinus of a 75-year old male patient was examined clinically, histologically, and by electron microscopy. The fine structure of neurinomas occurs in two forms: Type Antoni A is composed of Schwann cells whose nuclei are arranged in palisading rows with greatly attenuated cytoplasmic processes extending from the Schwann cells in parallel alignment; Antoni B is characterized by loosely arranged Schwann cells set in meshwork of macrocysts and reticular fibers. It may be a degenerative form of Antoni A. In the present case Antoni A was the dominating pattern. A prominent basal lamina enveloping Schwann cells, as well as desmosome-like junctions between them, were identified. Furthermore, myelin figures, lysosomal bodies, intranuclear unmyelinated axons and concentric laminated inclusions, and annulate lamellae were found in Schwann cells. Luse bodies appeared in the tumour matrix. It is suggested that annulate lamellae may play a significant role in cell differentiation and tumour growth. No other ultrastructural signs of malignancy were seen. PMID- 2554938 TI - [Experimental studies of the obliteration of extensive radical ear cavities with a new kind of ceramic granulate]. AB - Modern ear surgery aims at obliterating the extensive radical cavities that remain after removal of a cholesteatoma. A wide variety of techniques have been developed, including working with muscle fascial flaps, cartilage or bone chips; osteoplastic meato-atti++----+-coantrotomy; reconstruction of the posterior auditory duct wall by ceramic dish moulds or homologous cartilage with and without obliteration of the remaining mastoid cavity. The ceramic granulate available for obliteration of the mastoid cavity is responsible for extremely high costs. In an animal experimental study, we investigated the histocompatibility of several new ceramic substances. These new ceramic materials differ especially from the conventional granulate in their implantation behaviour and also in their long-term absorption behaviour, an advantage that enables this ceramic granulate to be used on a broad scale for obliteration of the mastoid cavity and thus the implementation of a largely physiological reconstruction. PMID- 2554939 TI - Molecular cloning and expression of Bacillus thuringiensis subsp. galleriae insecticidal crystal protein genes in Escherichia coli. AB - The location of the toxin gene of B. thuringiensis subsp. galleriae (H5ab) on the Mr-130Md plasmid is determined by molecular cloning. Double digestion fragments (BamHI and SalI) and PstI restriction fragments as well, from the 130 Md plasmid, of B. thuringiensis subsp. galleriae, are ligated with the cloning vector pAT 153 respectively and transformed into E. coli strain HB 101. Out of 208 transformants, three colonies (FG2, FG9, FG19) give positive hybridization reaction using the HD-1 delta-endotoxin gene as a probe. They are presumed to contain the delta-endotoxin gene of B. thuringiensis subsp. galleriae. Western blot assays indicate that Mr-130 kDal and 68 kDal, crystal proteins produced by clone FG2 react with anticrystal protein antibody. The protein extracts of clone FG2 are lethal to Ostrinia furnacalis (Guenee). This is the first report with regard to the cloning and expression of the B. thuringiensis subsp. galleriae (H5ab) delta-endotoxin gene. PMID- 2554940 TI - Forebrain monoamines and associative learning: III. The US pre-exposure effect. AB - This experiment evaluated the effects of forebrain norepinephrine (NE) and serotonin depletion on the unconditioned stimulus (US) pre-exposure phenomenon. Rats with either dorsal bundle lesions, raphe lesions, or which served as vehicle controls were assigned to one of two training conditions. One-half of the animals in the 3 lesion groups received 30 episodes of a stimulus which later served as the US. The remaining rats experienced standard conditioning of CS-US pairing without US pre-exposure. The results indicate that while raphe-lesioned and vehicular rats exhibited the US pre-exposure effect, conditioning was unimpaired in NE-depleted rats following pre-exposure. These findings are consistent with a contextual blocking interpretation of the US pre-exposure effect. PMID- 2554941 TI - FTIR study of netropsin binding to poly d(A-T) and poly dA.poly dT. AB - Complexes between netropsin and two polynucleotides containing only AT base pairs (poly d(A-T) and poly dA.poly dT) have been prepared at various drug/base pair ratios and studied in solution by Fourier Transform Infrared Spectroscopy. The drug is shown to interact in the narrow groove of poly d(A-T) with the C2O2 carbonyl of thymines and the N3 groups of adenines. Moreover the spectral modifications allow us to propose the existence of interactions at the level of the deoxyribose. No effect is detected on the phosphate groups when netropsin is progressively added. In the case of poly dA.poly dT the interaction seems much weaker as if the high propeller twist of the homopolymer would make the accessibility of the drug to the minor groove more difficult. PMID- 2554943 TI - Advances on beta-adrenergic receptors. Molecular structure and functional regulation. AB - The diverse effects of the catecholamines (CA), epinephrine and norepinephrine, are mediated by a family of specific receptors (adrenergic receptors, AR). The beta-AR is a glycoprotein present in the membrane of a number of cell types. This receptor is closely associated with at least two other proteins, guanine nucleotide stimulating protein (Gs) and adenylate cyclase enzyme (AC), to form the beta-AR complex. The beta-AR recognizes the CA and is coupled to Gs which stimulates the effector enzyme AC. This enzyme converts ATP to cAMP and is the effector of the beta-AR complex. Thus the beta-AR is a G-coupled receptor which acts by raising intracellular levels of cAMP. The beta-AR is an important site of regulatory modifications through a variety of mechanisms. The best characterized is known as homologous desensitization: when the receptor is exposed to repeated stimuli by the agonist (CA), its responsiveness wanes, probably to compensate this potentially dangerous overstimulation. The gene for mammalian beta 2-AR has recently been cloned and the predicted amino acid sequence now opens the field to identification of the protein structures involved in receptor functions. The beta 2-AR protein is characterized by the presence of seven membrane spanning regions. The study of the structure, function and regulation of the beta-AR will extend our knowledge of the role of beta-AR in pathological conditions and suggest new therapeutic approaches. PMID- 2554944 TI - Lewy bodies in the enteric nervous system in Parkinson's disease. AB - We systematically studied the intramural nervous system of the alimentary tract in patients with Parkinson's disease and found that Lewy bodies were distributed widely in the Auerbach's and Meissner's plexuses. In the central nervous system, we recognized a striking similarity between the distribution of Lewy bodies and that of monoaminergic neurons. More recently, we have demonstrated that neuronal somata immunoreactive for tyrosine hydroxylase (TH) exist in the Auerbach's and Meissner's plexuses of normal humans. We consider a possible relation between these TH-immunoreactive catecholaminergic neurons to the occurrence of Lewy bodies in the enteric nervous system in Parkinson's disease. The affinity of Lewy bodies to the central and enteric neurons seems to be attributable to an unknown cell-biological characteristic apparently shared by both neurons. PMID- 2554945 TI - Receptors of paraneurons, with special reference to glucoreceptors. AB - Among glucose-recognizing paraneurons, the gustatory cell is believed to have a glucoreceptor, while the pancreatic B cell is thought to metabolize glucose for signal production for insulin release. To investigate whether a common mechanism of glucose recognition exists among these cells, we have studied the interaction of the anomers of glucose or its derivative, known as a sugar taste inhibitor, with the pancreatic B cell of normal and diabetic rats. Inhibitors of the sugar taste response inhibited glucose-induced insulin release in various manners. A non-specific inhibitor, dibucaine, impaired not only the insulin response to glucose but also the ability to discriminate the anomers of glucose, without inhibiting glucose oxidation in the islets. Dibucaine also inhibited insulin release induced by a non-glucose secretagogue, tolbutamide. The alpha anomer, but not the beta anomer, of p-nitrophenyl-D-glucopyranoside, a specific-competitive inhibitor of sugar taste response, inhibited glucose-induced insulin release, but did not inhibit insulin release induced by non-glucose secretagogues or glucose oxidation. The pancreatic B cell of a non-insulin-dependent diabetes (NIDD) rat model, the NSZ rat, exhibited low insulin response to glucose and did not discriminate between the two anomers of glucose. The diabetic B cell responded to non-glucose secretagogues to the same extent as the control. Glucose oxidation in the diabetic islets was not impaired. These findings, together with previous ones, suggest that the gustatory and pancreatic B cells have a common glucose recognition site, which has a steric preference for the alpha anomer and is impaired in NIDD. PMID- 2554946 TI - Localization of glycine and beta-adrenergic receptors in the rat brain. AB - The localization of glycine (GR) and adrenergic receptors (AdR) was examined in the rat brain using a monoclonal antibody against the affinity purified glycine receptor and a polyclonal antibody against purified beta2-AdR. GR were concentrated in the lower brainstem, whereas no immunoreactivity was observed in the diencephalon and forebrain except in a few diencephalic nuclei. The highest density was found in the cranial motor nuclei, reticular formation, parabrachial area, dorsal and ventral cochlear nuclei, and dorsal and ventral tegmental nuclei. On the other hand, AdR were widely but unevenly distributed in the rat brain. The present study further showed the presence of AdR in catecholaminergic terminals in the hypothalamus. PMID- 2554942 TI - Patterns of steroid hormone effects on electrical and molecular events in hypothalamic neurons. AB - Hypothalamic neurons with nuclear receptors for steroid hormones provide opportunities to relate individual biosynthetic and electrical changes to hormone driven behaviors. Successful work with female rodent reproductive behavior has proven that it is possible to define a neural circuit for a vertebrate behavior. In contrast to what might be expected from an invertebrate system, results from several approaches to neuronal gene expression show the complexity of hypothalamic control, even over this simple mammalian behavior. This is not a 1 hormone-1 gene-1 behavior system. Neither is there just one mode of hormonal induction. Certain steroid hormone effects can multiply each other, showing how a clear endocrine signal could be discerned among other variations in neural activity. PMID- 2554947 TI - Fate of ATP in secretory granules: phosphohydrolase studies in pancreatic vascular bed. AB - Mature secretory granules in paraneurons contain ATP amongst other small messenger molecules. In the islet organ such stores of adenine nucleotides readily can be demonstrated by means of the quinacrine fluorescence method. ATP is co-released together with other granule constituents when the major hormones are exocytosed. The distribution of ATP splitting enzymic activities was studied in the pancreas of the mouse and rat, in order to obtain information on the possible fate of this small messenger molecule. ATPase, ADPase, and AMPase (5' nucleotidase) were demonstrated with lead precipitation methods, L-tetramisole was used to inhibit unspecific alkaline phosphatase (alPase); alPase activities were shown with tetrazolium methods, using 5-bromo-4-chloro-3-indoxyl phosphate as substrate. Most endothelial cells of the vascular bed, both in the exocrine and in the endocrine pancreas, are reactive for ATPase, ADPase, AMPase and alPase. Smooth muscle cells are strongly reactive for ATPase and AMPase, vascular adventitial fibroblasts (veil cells) stain for ATPase and alPase, as do some lamellar cells at the islets surface. Staining for ADPase serves as a selective method to demonstrate the vascular bed. Comparable results are obtained with the alPase reaction, though insular non-B-cells are also reactive. ATPase staining is less useful for demonstrating vascular connections because moderate reactivity of exocrine parenchyma and adventitial tissue obscures the picture. AMPase activity is strong in the venous segments of the capillary net and in collecting veins but the reaction obviously does not demonstrate significant portions of the residual capillary network. Weak AMPase activity is seen in the insular parenchyma.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554948 TI - [Expression of insulin-like growth factor I receptors in human brain tumors: comparison with epidermal growth factor receptor by using quantitative autoradiography]. AB - By using quantitative autoradiographic techniques, receptors for insulin-like growth factor I (IGF-I) and epidermal growth factor (EGF) were analyzed in 13 samples of human brain tumors (4 low grade astrocytomas, 7 glioblastomas, 1 anaplastic ependymoma and 1 medulloblastoma). High number of specific binding sites for IGF-I and EGF were homogeneously present in tissue sections derived from glioblastoma. In low grade astrocytoma, relatively high numbers of binding site for EGF were observed, but there was no significant difference in concentrations of IGF-I binding sites between tumors and control cortex. In medulloblastoma, only IGF-I binding sites were present. These observations might indicate that both IGF-I and EGF are involved in the growth modulation of human gliomas possibly through paracrine or autocrine mechanisms. Antagonists to growth factors or monoclonal antibody against those receptors could have the way for therapeutic application for gliomas. PMID- 2554949 TI - The relationship between ex vivo anti-factor Xa levels and efficacy/safety of low molecular weight heparin. PMID- 2554950 TI - Prevention of thromboembolic disease in general surgery with enoxaparin. PMID- 2554951 TI - Plasma from patients with severe Lassa fever profoundly modulates f-met-leu-phe induced superoxide generation in neutrophils. AB - A recurrent theme in studies of the pathology of fatal Lassa fever in man is the lack of histological lesions to explain disordered cell function and death. Recently, we demonstrated the existence of a factor in the plasma of patients with Lassa fever which markedly inhibits the aggregation responses of normal platelets in vitro. To assess whether this factor could mediate more global cellular dysfunction, we studied the effects of Lassa plasma on the respiratory burst of neutrophils. Thirteen of 15 samples from patients in the acute phase of Lassa fever profoundly inhibited the amount of superoxide generated by normal neutrophils in response to the chemotactic peptide, f-met-leu-phe (FMLP) (mean superoxide generated = 54.7 +/- 6.1% of control). In contrast, eight of nine samples from patients who had infections other than Lassa fever enhanced the neutrophil response to the peptide. All Lassa samples which inhibited the ADP induced aggregation responses of normal platelets inhibited the neutrophil response to FMLP. Unlike the effect on platelets, however, the inhibition of neutrophils was only apparent when the cells were stimulated within 5 min of exposure to the plasma. The inhibition of neutrophils is not due to either interference with FMLP-neutrophil binding or an effect on the NADPH-oxidase, suggesting a suppression of signal transduction. Our data suggest the inhibitory factor in Lassa plasma has global effects on cellular function, and may play a central role in the pathogenesis of this often fatal illness. PMID- 2554952 TI - The use of DNA probes to monitor minimal residual disease in childhood acute lymphoblastic leukaemia. AB - DNA probes to both the joining region (JH) of the immunoglobulin heavy chain gene (IgH) and to the beta chain of the T-cell antigen receptor complex (TCR) have been used as tumour-specific markers to monitor the rearrangements of the IgH chain gene and the TCR beta gene in the blast cells of children presenting with acute lymphoblastic leukaemia (ALL) of B or T cell origin. Blast cells from 68 children with early B cell ALL and eight with T-ALL were examined at presentation, at day 28 after commencement of therapy and at varying times thereafter. An additional 43 patients (42 with B cell ALL, one with T-ALL) were studied both at presentation, at completion of their 2-year treatment course and 3 months later. Twelve patients, drawn from both these groups, were studied at relapse as were a further eight patients in whom an extramedullary relapse had occurred. Persistence of clonally-derived cells as a predictor of early relapse was seen in the day 28 bone marrows of 11/76 newly-diagnosed children (nine early B and two T-ALL) followed by rapid, overt relapse in four of the early B ALL cases. No minimal residual disease (MRD) was detected in bone marrows from any of the 43 patients completing their 2-year treatment course, but six of these subsequently relapsed at varying time periods thereafter. Identical patterns of rearrangement at both presentation and relapse were seen in most cases. Oligoclonality, or multiple IgH chain gene rearrangements was seen in the blast cells of 15% of patients with early B cell ALL. No correlation between oligoclonality, high white count, unfavourable phenotype or abnormal karyotype could, however, be ascertained. PMID- 2554953 TI - Identification of H- and L-ferritin subunit binding sites on human T and B lymphoid cells. AB - Monoclonal antibodies to the H and L subunits of ferritin were used in indirect immunofluorescent studies to investigate the subunit specificity of ferritin binding to transformed T and B lymphoid cell lines. Pre-incubation of cells with recombinant H ferritin followed by antibodies to the H subunit produced a labelling pattern in which all cells showed strong fluorescent stippling. Corresponding experiments with liver ferritin and an antibody to the L subunit revealed that most cells were lightly stippled, but 3-5% of the cells were intensely labelled. These two labelling patterns were distinct as assessed by cross-blocking experiments with recombinant H and liver ferritins. These results imply that different binding sites exist for each subunit. PMID- 2554954 TI - Antibody producing cells in the spleens of mice treated with pathogenic mineral dust. AB - Experiments were carried out to assess the effect of intraperitoneal injection of the mineral dusts, titanium dioxide, quartz, or asbestos, on splenic lymphocyte antibody forming cells in immunised mice. Titanium dioxide and quartz caused similar, about one third, reductions in plaque forming cells; asbestos caused substantial reduction to about a quarter of the number found in control spleens. The inhibition of antibody forming cells in the spleen found with chrysotile was dose dependent and both chrysotile and crocidolite asbestos were similar in activity. Systemic immunomodulation after local deposition of mineral dust may be important to the development of disease. PMID- 2554955 TI - Laboratory studies of the dental properties of soft drinks. AB - The composition and dental properties of eight different soft drinks, representing some of the most popular types used in the UK, were examined. Demineralization experiments were conducted on hydroxylapatite, the basic component of dental enamel, determining calcium dissolving by atomic absorption spectroscopy and phosphorus by u.v. visual spectrophotometry. The titratable acid content of the drinks was found to give a better guide than their pH to their potential dental erosiveness. The sugar content, in their ready-to-drink form, varied from zero in a low-calorie product up to almost 14% in a blackcurrant drink, but using a technique with a relatively long contact time, and in the absence of intact dental plaque, the demineralizing action on hydroxylapatite of the acids already in the drinks eclipsed the effects of the acid generated by oral micro-organisms from the sugars in the drinks. The pure citrus juices showed potentially the worst dental properties, followed by the orange and blackcurrant concentrates after dilution to their ready-to-drink form, with least demineralization from the carbonated drinks, and a cola drink giving especially low values. PMID- 2554957 TI - Acid dissociation constant and apparent nucleophilicity of lysine-501 of the alpha-polypeptide of sodium and potassium ion activated adenosinetriphosphatase. AB - A combination of competitive labeling with [3H]acetic anhydride [Kaplan, H., Stevenson, K. J., & Hartley, B. S. (1971) Biochem. J. 124, 289-299] and immunoaffinity chromatography is described that permits the assignment of the acid dissociation constant and the absolute nucleophilicity of individual lysines in a native enzyme. The acid dissociation constant of lysine-501 of the alpha polypeptide in native (Na+ + K+)-ATPase was determined. This lysine had a normal pKa of 10.4. The rate constant for the reaction of the free base of lysine-501 with acetic anhydride at 10 degrees C is 400 M-1 s-1. This value is only 30% that for a fully accessible lysine in a protein. The lower than normal apparent nucleophilicity suggests that lysine-501 is hindered from reacting with its intrinsic nucleophilicity by the tertiary structure of the enzyme and is consistent with its location within a pocket that forms the active site upon the surface of the native protein. PMID- 2554956 TI - Proteolytic modification of calcium-dependent protease 1 in erythrocytes treated with ionomycin and calcium. AB - In vitro, limited proteolytic cleavage of the subunits of the purified calcium dependent proteases [also known as calpains (EC 3.4.22.17) or calcium-activated neutral proteinases (CANPs)] appears to be required for enzyme activity. It has not yet been demonstrated if similar processing of the protease subunits occurs in vivo. To directly assess proteolytic modification of these proteases in cells, we have measured the loss of the proenzyme form of the regulatory subunit (a 26 kDa protein) and/or the appearance of the modified regulatory subunit (a 17-kDa protein) by densitometric analysis of immunoblots. In rat erythrocytes, proteolytic modification of the endogenous calcium-dependent protease (calcium dependent protease 1, mu CANP) occurs in vivo in response to ionomycin and calcium. The extent of enzyme modification was dependent on time, ionomycin concentration, and calcium concentration, suggesting that in this cellular model Ca2+ regulates proteolytic modification of the enzyme. PMID- 2554958 TI - Evidence from 18O exchange measurements for steps involving a weak acid and a slow chemical transformation in the mechanism of phosphorylation of the gastric H+, K+-ATPase by inorganic phosphate. AB - Phosphorylation of the gastric H,K-ATPase by Pi has been studied by measuring the P18Oj16O4-j distribution as a function of time at different H+, K+, and [18O]Pi concentrations. The advantage of isotope exchange measurements is that the P18Oj16O4-j distribution depends on the relative rates of HOH loss to form the phosphoenzyme intermediate and Pi dissociation from the enzyme. Therefore, 18O exchange is a sensitive probe of mechanism. K+ increases the exchange rate (v(ex] but does not affect the partition coefficient (Pc) that determines the P18Oj16O4 j distribution. Conversely, H+ inhibits exchange. A single Pc describes the data at every pH, but the value increases from 0.04 at pH 8 to 0.64 at pH 5.5. Vex depends hyperbolically on [Pi]0. Km for Pi does not depend on pH, and Pc does not depend on [Pi]0. Individual rate constants in the phosphorylation mechanism are estimated. Formation of the E.Pi complex that looses HOH is 1-2 orders of magnitude slower at pH 5.5 than at pH 8 and is not diffusion controlled. The observed change in Pc with pH is compatible with catalysis occurring by a different mechanism when a group with pKa = 7.2 is protonated. Slower than diffusion-controlled formation of the E.Pi complex that splits out HOH is evidence for a relatively slow, unimolecular chemical transformation involving an additional intermediate in the phosphorylation mechanism, such as a protein conformational change. PMID- 2554959 TI - Escherichia coli cAMP receptor protein: evidence for three protein conformational states with different promoter binding affinities. AB - Cyclic AMP receptor protein (CRP) from Escherichia coli is assumed to exist in two states, namely, those represented by the free protein and that of the ligand protein complex. To establish a quantitative structure-function relation between cAMP binding and the cAMP-induced conformational changes in the receptor, protein conformational change was quantitated as a function of cAMP concentration up to 10 mM. The protein conformation was monitored by four different methods at pH 7.8 and 23 degrees C, namely, rate of proteolytic digestion by subtilisin, rate of chemical modification of Cys-178, tryptophan fluorescence, and fluorescence of the extrinsic fluorescence probe 8-anilino-1-naphthalenesulfonic acid (ANS). Each of these techniques reveals a biphasic dependence of protein conformation on cAMP concentration. At low cAMP concentrations ranging from 0 to 200 microM, the rates of proteolytic digestion and that of Cys-178 modification increase, whereas the fluorescence intensity of the ANS-protein complex is quenched, and there is no change in the fluorescence intensity of the tryptophan residues in the protein. At higher cAMP concentrations, the rates of proteolytic and chemical modification of the protein decrease, while the fluorescence intensity of the ANS-protein complex is further quenched but there is an increase in the intensity of tryptophan fluorescence. These results show unequivocally that there are at least three conformational states of the protein. The association constants for the formation of CRP-cAMP and CRP-(cAMP)2 complexes derived from conformational studies are in good agreement with those determined by equilibrium dialysis, nonequilibrium dialysis, and ultrafiltration. Therefore, the simplest explanation would be that the protein exhibits three conformational states, free CRP and two cAMP-dependent states, which correspond to the CRP-cAMP and CRP-(cAMP)2 complexes. The binding properties of CRP-cAMP and CRP-(cAMP)2 to the lac promoter were studied by using the gel retardation technique. At a high concentration of cAMP which favors the formation of the CRP-(cAMP)2 complex, binding of the protein to DNA is decreased. This, together with conformational data, strongly suggests that only the CRP-cAMP complex is active in specific DNA binding whereas CRP and CRP-(cAMP)2 are not. PMID- 2554960 TI - Affinity labeling of the folate-methotrexate transporter from Leishmania donovani. AB - An affinity labeling technique has been developed to identify the folate methotrexate transporter of Leishmania donovani promastigotes using "activated" derivatives of the ligands. These "activated" derivatives were synthesized by incubating folate and methotrexate with a 10-fold excess of 1-ethyl-3-[3 (dimethylamino)propyl]carbodiimide (EDC) for 10 min at ambient temperature in dimethyl sulfoxide. Preincubation of intact cells with nonradioactive "activated" folate or methotrexate at a concentration of 40 microM inhibited the capacity of wild-type cells to transport submicromolar concentrations of unmodified ligand. When intact wild-type (DI700) Leishmania donovani or preparations of their membranes were incubated with a 0.4 microM concentration of either "activated" [3H]folate or "activated" [3H]methotrexate, the radiolabeled ligands were covalently incorporated into a polypeptide with a molecular weight of approximately 46,000, as demonstrated by SDS-polyacrylamide gel electrophoresis. No affinity labeling of a 46,000-dalton protein was observed when equimolar concentrations of "activated" radiolabeled ligands were incubated with intact cells or membranes prepared from a methotrexate-resistant mutant clone of Leishmania donovani, MTXA5, that is genetically defective in folate-methotrexate transport capability [Kaur, K., Coons, T., Emmett, K., & Ullman, B. (1988) J. Biol. Chem. 263, 7020-7028]. However, some labeling of a 46,000-dalton protein was observed when MTXA5 cells were incubated with higher concentrations of "activated" ligands. Time course studies indicated that maximal labeling of the 46,000-dalton protein occurred within 5-10 min of incubation of intact cells with "activated" ligand.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554961 TI - Role of specific lysine residues in binding cytochrome c2 to the Rhodobacter sphaeroides reaction center in optimal orientation for rapid electron transfer. AB - The role of specific lysine residues in facilitating electron transfer from Rhodobacter sphaeroides cytochrome c2 to the Rb. sphaeroides reaction center was studied by using six cytochrome c2 derivatives each labeled at a single lysine residue with a carboxydinitrophenyl group. The reaction of native cytochrome c2 at low ionic strength has a fast phase with a half-time of 0.6 microseconds that has been assigned to the reaction of bound cytochrome c2 [Overfield, R.E., Wraight, C.A., & DeVault, D. (1979) FEBS Lett. 105, 137]. Modification of lysine 55 did not affect the half-time of this phase but decreased the apparent binding constant by a factor of 2. The derivatives modified at lysines-10, -88, -95, -97, -99, -105, and -106 surrounding the heme crevice did not show any detectable fast phase but only slow second-order phases due to the reaction of solution cytochrome c2. These lysines thus appear to be involved in binding cytochrome c2 to the reaction center in an optimal orientation for electron transfer. The involvement of lysines-95 and -97 is especially significant, since they are located in an extra loop comprising residues 89-98 that is not present in eukaryotic cytochrome c. The reactions of horse cytochrome c derivatives modified at single lysine amino groups with trifluoroacetyl or [(trifluoromethyl)phenyl]carbamoyl were also studied. The derivatives modified at lysines-22, -55, -88, and -99 far removed from the heme crevice had nearly the same half-times for the fast phase as native cytochrome c, 6 microseconds.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554962 TI - Electron transfer between cytochrome a and copper A in cytochrome c oxidase: a perturbed equilibrium study. AB - Intramolecular electron transfer in partially reduced cytochrome c oxidase has been studied by the perturbed equilibrium method. We have prepared a three electron-reduced, CO-inhibited form of the enzyme in which cytochrome a and copper A are partially reduced and in an intramolecular redox equilibrium. When these samples were irradiated with a nitrogen laser (0.6-ns, 1.0-mJ pulses) to photodissociate the bound CO, changes in absorbance at 598 and 830 nm were observed which were consistent with a fast electron transfer from cytochrome a to copper A. The absorbance changes at 598 nm gave an apparent rate of 17,000 +/- 2000 s-1 (1 sigma), at pH 7.0 and 25.5 degrees C. These changes were not observed in either the CO mixed-valence or the CO-inhibited fully reduced forms of the enzyme. The rate was fastest at about pH 8.0, falling off toward both lower and higher pHs. There was a small but clear temperature dependence. The process was also observed in the cytochrome c-cytochrome c oxidase high-affinity complex. The electron equilibration measured between cytochrome a and copper A is far faster than any rate measured or inferred previously for this process. PMID- 2554963 TI - A nuclear Overhauser effect investigation of the molecular and electronic structure of the heme crevice in lactoperoxidase. AB - The proton homonuclear nuclear Overhauser effect, NOE, in conjunction with paramagnetic-induced dipolar relaxation, is utilized to assign resonances and to probe the molecular and electronic structures of the heme cavity in the low-spin cyanide complex of resting-state bovine lactoperoxidase, LPO-CN. Predominantly primary NOEs were detected in spite of the large molecular weight (approximately 78 x 10(3)) of the enzyme, which demonstrates again the advantage of paramagnetism suppressing spin diffusion in large proteins. Both of the nonlabile ring protons of a coordinated histidine are located at resonance positions consistent with a deprotonated imidazole. Several methylene proton pairs are identified, of which the most strongly hyperfine-shifted pair is assigned to the unusual chemically functionalized 8-(mercaptomethylene) group of the prosthetic group [Nichol, A. W., Angel, L. A., Moon, T., & Clezy, P. S. (1987) Biochem. J. 247, 147-150]. The large 8-(mercaptomethylene) proton contact shifts relative to that of the only resolved heme methyl signal are rationalized by the additive perturbations on the rhombic asymmetry of the functionalization of the 8-position and the alignment of the axial histidyl imidazole projection along a vector passing through pyrrole A and C of the prosthetic group. Such a stereochemistry is consistent with the resolution of only a single heme methyl group, 3-CH3, as observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554965 TI - The sodium ion translocating adenosinetriphosphatase of Propionigenium modestum pumps protons at low sodium ion concentrations. AB - The purified ATPase (F1F0) of Propionigenium modestum has its pH optimum at pH 7.0 or at pH 6.0 in the presence or absence of 5 mM NaCl, respectively. The activation by 5 mM NaCl was 12-fold at pH 7.0, 3.5-fold at pH 6.0, and 1.5-fold at pH 5.0. In addition to its function as a primary Na+ pump, the ATPase was capable of pumping protons. This activity was demonstrated with reconstituted proteoliposomes by the ATP-dependent quenching of the fluorescence of 9-amino-6 chloro-2-methoxyacridine. No delta pH was formed in the presence of the uncoupler carbonyl cyanide m-chlorophenylhydrazone or by blocking the ATPase with dicyclohexylcarbodiimide. In the presence of valinomycin and K+, the delta pH increased, in accord with the operation of an electrogenic proton pump. The proton pump was only operative at low Na+ concentrations (less than 1 mM), and its activity increased as the Na+ concentration decreased. Parallel to the decrease of H+ pumping, the velocity of the Na+ transport increased about 6-fold from 0.1 to 4 mM NaCl, indicating a switch from H+ to Na+ pumping, as the Na+ concentration increases. Due to proton leaks in the proteoliposomal membranes, fluorescence quenching was released after blocking the ATPase with dicyclohexylcarbodiimide, by trapping residual ATP with glucose and hexokinase, or by the Na+-induced conversion of the proton pump onto a Na+ pump. Amiloride, an inhibitor of various Na+-coupled transport systems, was without effect on the kinetics of Na+ transport by the P. modestum ATPase. PMID- 2554964 TI - Activation of O2.- generating oxidase of bovine neutrophils in a cell-free system. Interaction of a cytosolic factor with the plasma membrane and control by G nucleotides. AB - Activation of the O2.- -generating oxidase of bovine neutrophils was studied in a cell-free system, consisting of a particulate fraction enriched in plasma membrane, cytosol, arachidonic acid, and the non-hydrolyzable nucleotide GTP gamma-S. Activation of the membrane-bound oxidase was accompanied by the disappearance of the activating factor from the cytosol. Above a cytosol to membrane ratio of 25, the excess of added cytosolic factor remained in active state in the soluble fraction. The process could be partially reversed by serum albumin. Disappearance of the cytosolic factor was promoted by unsaturated long chain fatty acids, but not by saturated ones, and occurred not only in the presence of GTP-gamma-S but also in the presence of GDP-beta-S or in the absence of Mg ions, although in the latter cases activation of O2.- production was seriously impaired. This suggests that the disappearance of the activating factor from the cytosol and the triggering effect of GTP-gamma-S are related, but distinct, events in the oxidase activation process. The disappearance of the activating factor from cytosol can be explained by translocation of the cytosolic factor to the membrane fraction. Yet under some conditions, including the presence of GDP-beta-S or EDTA, inactivation was prevailing and could be an alternative explanation for the results. Specific binding of radiolabeled GTP gamma-S could be demonstrated both in the membrane and in the cytosolic fractions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554966 TI - Proton-metal distance determination in cobalt(II) stellacyanin by 1H nuclear magnetic resonance relaxation measurements including Curie-spin effects: a proposed structure of the metal-binding region. AB - 1H nuclear magnetic resonance (1H NMR) experiments on Co(II)-substituted stellacyanin have been performed. Large paramagnetic hyperfine shifts are observed, the whole spectrum covering a range of 190 ppm. Experiments were mainly performed at 270 MHz from which temperature and pH* dependencies of the out shifted resonances were reported, as well as determinations of the longitudinal (T1) and transverse (T2) relaxation times. These relaxation times are among other things, dependent on the individual proton-metal distance, and the aim of this work has been to determine these distances, by use of the Solomon-Bloembergen equations modified to include the so-called "Curie spin". The application of this method to a protein has not been reported earlier. Experiments were also performed at 100, 400, and 500 MHz in order to estimate the size of the Curie spin from the field dependence of the line widths. Furthermore, determination of the values for the rotational correlation time, tau r, and the effective magnetic moment, mu eff, was necessary for the present approach. With apostellacyanin, tau r was found to be (6.0 +/- 0.4) X 10-8 s. From the paramagnetic susceptibility of Co(II) stellacyanin, the value (4.53 +/- 0.03)beta was determined for mu eff. The proposed assignments of several paramagnetically out-shifted resonances. the proton-metal distances obtained, and the known peptide sequence of stellacyanin have allowed us to build a three-dimensional model of the metal site and its surrounding structure consistent with all the experimental data. It is revealed that both histidine ligands bind the metal with their 3-nitrogens. Also we find strong indications that a second sulfur atom is actually binding the metal, this being the long-sought-after fourth ligand. The model suggests that this sulfur belongs to Cys-59, which together with Cys-93 constitutes the disulfide bridge known to be present in the structure. A potential fifth ligand, an amide oxygen from Asn-47, is also found. PMID- 2554967 TI - Determination of the three-dimensional solution structure of the C-terminal domain of cellobiohydrolase I from Trichoderma reesei. A study using nuclear magnetic resonance and hybrid distance geometry-dynamical simulated annealing. AB - The solution structure of a synthetic 36-residue polypeptide comprising the C terminal cellulose binding domain of cellobiohydrolase I (CT-CBH I) from Trichoderma reesei was investigated by nuclear magnetic resonance (NMR) spectroscopy. The 1H NMR spectrum was completely assigned in a sequential manner by two-dimensional NMR techniques. A large number of stereospecific assignments for beta-methylene protons, as well as ranges for the phi, psi, and chi 1 torsion angles, were obtained on the basis of sequential and intraresidue nuclear Overhauser enhancement (NOE) and coupling constant data in combination with a conformational data base search. The structure calculations were carried out in an iterative manner by using the hybrid distance geometry-dynamical simulated annealing method. This involved computing a series of initial structures from a subset of the experimental data in order to resolve ambiguities in the assignments of some NOE cross-peaks arising from chemical shift degeneracy. Additionally, this permitted us to extend the stereospecific assignments to the alpha-methylene protons of glycine using information on phi torsion angles derived from the initial structure calculations. The final experimental data set consisted of 554 interproton distance restraints, 24 restraints for 12 hydrogen bonds, and 33 phi, 24 psi, and 25 chi 1 torsion angle restraints. CT-CBH I has two disulfide bridges whose pairing was previously unknown. Analysis of structures calculated with all three possible combinations of disulfide bonds, as well as without disulfide bonds, indicated that the correct disulfide bridge pairing was 8-25 and 19-35. Forty-one structures were computed with the 8-25 and 19-35 disulfide bridges, and the average atomic rms difference between the individual structures and the mean structure obtained by averaging their coordinates was 0.33 +/- 0.04 A for the backbone atoms and 0.52 +/- 0.06 A for all atoms. The protein has a wedgelike shape with an amphiphilic character, one face being predominantly hydrophilic and the other mainly hydrophobic. The principal element of secondary structure is made up of an irregular triple stranded antiparallel beta-sheet composed of residues 5-9 (beta 1), 24-28 (beta 2), and 33-36 (beta 3) in which strand beta 3 is hydrogen bonded to the other two strands.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2554969 TI - Delta opioid receptor selectivity induced by conformational constraints in linear enkephalin-related peptides: 1H 400-MHz NMR study and theoretical calculations. AB - Introduction into the structure of the linear hexapeptide DSLET (Tyr-D-Ser-Gly Phe-Leu-Thr) or DTLET (Tyr-D-Thr-Gly-Phe-Leu-Thr) of tert-butyl groups as constraints different from cyclization leads to a large increase in the selectivity for delta opioid binding site in the case of DSTBULET [Tyr-D-Ser (OtBu)-Gly-Phe-Leu-Thr] (Ki delta = 6.14 nM; Ki mu = 374 nM) and BUBU [Tyr-D Ser(OtBu)-Gly-Phe-Leu-Thr(OtBu)] (Ki delta = 4.68 nM; Ki mu = 475 nM) or a loss of affinity for DTTBULET [Tyr-D-Thr(OtBu)-Gly-Phe-Leu-Thr] (Ki delta = 866 nM; Ki mu = 4500 nM). This puzzling behavior is studied here by 400-MHz 1H NMR spectroscopy in DMSO-d6 solution and by theoretical calculations. When DSLET and DTLET are compared, the reduction in energetically accessible phi and psi angles induced by the tert-butyl group in the D-Ser2 residue decreases the degree of freedom in the N-terminal part of the peptides. For DSTBULET and BUBU, the rigidification of the backbone evidenced by the appearance of the large NOE's of Phe4 NH-Gly3 alpha and Gly3 NH-alpha and by the loss of the C7 folding around the D-Ser2 residue found in DSLET could explain the drastic loss of affinity for mu opioid receptors. In DTTBULET, a large change in the spatial orientation around the D-Thr2 (OtBu) residue forces the aromatic rings far from each other.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2554968 TI - Synergistic release of arachidonic acid from platelets by activators of protein kinase C and Ca2+ ionophores. Evidence for the role of protein phosphorylation in the activation of phospholipase A2 and independence from the Na+/H+ exchanger. AB - The protein kinase C activators phorbol myristate acetate (PMA), mezerein, oleoylacetylglycerol, and (-)-indolactam V, although without direct effect on arachidonic acid release, greatly enhance the release of platelet arachidonic acid caused by the Ca2+ ionophores A23187 and ionomycin. In contrast, 4 alpha phorbol 12,13-didecanoate and (+)-indolactam V, which lack the ability to activate kinase C, do not potentiate arachidonate release. Release of arachidonic acid occurs without activation of phospholipase C and is therefore mediated by phospholipase A2. Synergism between PMA and A23187 is not affected by inactivation of the Na+/H+ exchanger with dimethylamiloride. The time course and dose-response for the effect of PMA at 23 degrees C closely correlate with the phosphorylation of a set of relatively "slowly" phosphorylated proteins (P20, P35, P41, P60), but not the rapidly phosphorylated P47 protein. P20 is myosin light chain, and P41 is probably Gi alpha, but the other proteins have not been positively identified. Depletion of metabolic ATP stores by antimycin A plus 2 deoxyglucose abolishes both protein phorphorylation and the potentiation of arachidonate release by PMA, but does not prevent fatty acid release by the ionophores. Similarly, the kinase C inhibitors H-7 and staurosporine produce, respectively, partial and complete inhibition of PMA-potentiated arachidonic acid release and protein phosphorylation, without affecting the direct response to ionophores. These results indicate that protein phosphorylation, mediated by kinase C, promotes the phospholipase A2 dependent release of arachidonic acid in platelets when intracellular Ca2+ is elevated by Ca2+ ionophores. PMID- 2554970 TI - Activation of cGMP phosphodiesterase in retinal rods: mechanism of interaction with the GTP-binding protein (transducin). AB - The mechanism of activation of cGMP phosphodiesterase by the GTP-binding protein in the disc membrane of retinal rods has been investigated by measuring the light induced phosphodiesterase activity in reconstituted systems where the concentration of either the GTP-binding protein or the phosphodiesterase is varied. The results are consistent with the existence of two activator sites per phosphodiesterase functional unit: binding of one G alpha GTP (alpha subunit of the G-protein with GTP bound) with high affinity (100 +/- 50 nM) partially activates the enzyme (Vmax1 approxmately 0.05 Vmax to 0.10V max to trypsin activated phosphodiesterase); binding of a second G alpha GTP with lower affinity (600 +/- 100 nM) induces maximal activation (Vmax2 approximately Vmax of trypsin activated phosphodiesterase). The two different states of activated phosphodiesterase have the same Km for cGMP and the same pH dependence; they differ in their sensitivity to GMP. Micromolar concentration of protamines increases the affinity of the two activator sites and slightly increases Vmax1. When G-protein is activated with GTP-gamma S instead of GTP, the affinities of the two activator sites are not significantly modified, while Vmax1 appears to be increased. PMID- 2554972 TI - Type I collagen degradation by mouse calvarial osteoblasts stimulated with 1,25 dihydroxyvitamin D-3: evidence for a plasminogen-plasmin-metalloproteinase activation cascade. AB - To understand the mechanisms regulating osteoid removal by osteoblasts, mouse calvarial osteoblasts were grown on 14C-labelled type I collagen films and stimulated with 1,25-dihydroxyvitamin D-3 (2.5.10(-8) M) for 48-72 h. In the presence of 5% non-inhibitory rabbit serum this resulted in a 2-3-fold increase in collagen degradation and a dramatic change in osteoblast morphology, when compared with untreated osteoblasts. Collagenolysis was accompanied by increased synthesis and release of latent collagenase, gelatinase and stromelysin and a concomitant decrease in their specific inhibitor, TIMP (tissue inhibitor of metalloproteinases). In serum-free medium, osteoblasts failed to degrade collagen, but their ability to lyse collagen could be restored by adding plasminogen (5 micrograms/ml) to the cultures. Plasminogen-dependent collagenolysis was inhibited by human recombinant TIMP (5 units/ml), demonstrating that plasmin, derived from plasminogen, activated latent collagenase and did not itself degrade collagen. Plasminogen activator production was confirmed by culturing osteoblasts on 125I-labelled fibrin plates. Comparison with urokinase-type and tissue-type plasminogen activator standards suggested that osteoblast plasminogen activator was predominantly cell-associated and likely to be of the urokinase type. Immunocytochemistry indicated that osteoblasts also constitutively produce plasminogen activator inhibitor-1. These findings provide evidence for the involvement of a plasminogen-plasmin-latent metalloproteinase activation cascade in type I collagen degradation by osteoblasts, and for its regulation by TIMP and plasminogen activator inhibitor 1. PMID- 2554971 TI - Spin-label ESR studies of lipid-protein interactions in thylakoid membranes. AB - Lipid-protein interactions in thylakoid membranes, and in the subthylakoid membrane fractions containing either photosystem 1 or photosystem 2, have been studied by using spin-labeled analogues of the thylakoid membrane lipid components, monogalactosyldiacylglycerol, phosphatidylglycerol, and phosphatidylcholine. The electron spin resonance spectra of the spin-labeled lipids all consist of two components, one corresponding to the fluid lipid environment in the membranes and the other to the motionally restricted membrane lipids interacting directly with the integral membrane proteins. Spectral subtraction has been used to quantitate the fraction of the membrane lipids in contact with the membrane proteins and to determine the selectivity between the different lipid classes for the lipid-protein interaction. The fractions of motionally restricted lipid in the thylakoid membrane are 0.36, 0.39, and 0.53, for the spin-labeled monogalactosyldiacylglycerol, phosphatidylcholine, the phosphatidylglycerol, respectively. Spin-labeled monogalactosyldiacylglycerol exhibits very little preferential interaction over phosphatidylchline, which suggests that part of the role of monogalactosyldiacylglycerol in thylakoid membranes is structural, as is the case for phosphatidylcholine in mammalian membranes. Spin-labeled phosphatidylglycerol shows a preferential interaction over the corresponding monogalactosyldiacylglycerol and phosphatidylcholine analogues, in contrast to the common behavior of this lipid in mammalian systems. This pattern of lipid selectivity is preserved in both the photosystem 1 and photosystem 2 enriched subthylakoid membrane fractions. PMID- 2554973 TI - Oximetry in cells and tissues using a nitroxide-liposome system. AB - In order to avoid the complication of reduction of nitroxides in biological media during oxygen measurements, liposomes containing a water-soluble nitroxide, 2,2,6,6-tetramethyl-piperidine-N-oxyl-4-trimethylammonium (Cat1), were used in studies of oxygen consumption by thymus-bone-marrow cells. The superhyperfine structure of Cat1 contained in liposomes was found to be sensitive to oxygen concentration in a fashion similar to that of free Cat1. Measurements of cellular respiration using Cat1 contained in liposomes agreed well with the results obtained using free Cat1. Using this nitroxide-liposome system, the respiration of liver slices was measured successfully, whereas such measurements using free cat1 were complicated by rapid reduction of the nitroxide. This nitroxide liposome system also could be used in conjunction with a membrane permeable nitroxide and an extracellular broadening agent to measure intracellular and extracellular oxygen concentrations simultaneously. PMID- 2554974 TI - Interleukin-2- and phytohemagglutinin-activated proliferation of human T lymphocytes is accompanied by stimulation of phosphoinositide turnover. AB - Interleukin-2 (IL-2) was more effective than phytohemagglutinin (PHA) in increasing the proliferative activity of human T-lymphocytes. Unlike PHA, IL-2 stimulated phosphoinositide turnover (PI turnover) only in those T-lymphocytes which had been preactivated by PHA with IL-2 and expressed the IL-2 receptors. The effect of PHA on PI turnover was, in general, stronger than that of IL-2. These results indicate that IL-2 and PHA-activated proliferation of human T lymphocytes is accompanied by stimulation of PI turnover. However, IL-2-induced proliferative response may not be a direct consequence of PI turnover stimulation in these cells. PMID- 2554975 TI - Phosphatidylinositol-specific phospholipase C releases lipoprotein lipase from the heparin releasable pool in rat heart cell cultures. AB - The effect of phosphatidylinositol-specific phospholipase C (PI-PLC) on the release of lipoprotein lipase was studied in F1 heart cell cultures. Exposure of the cultures for 10 min to PI-PLC resulted in a 2-fold increase in the release of lipoprotein lipase (LPL) into the culture medium. PI-PLC released LPL from the heparin-releasable pool and PI-PLC was not effective in cultures pretreated with heparin. Insulin had no influence on the release of LPL from the heart cell cultures, even though it enhanced the uptake of 2-deoxy[3H]glucose by these cells. In cultures labeled with 35S, treatment with PI-PLC resulted in an increase in the release of 35S-labeled proteoglycan. PI-PLC was also effective in enhancing the release of bovine LPL exogenously bound to cultured aortic smooth muscle cells. The findings that PI-PLC was not effective after heparin, that it did release exogenously added LPL to cell cultures and that it released 35S labeled proteoglycan, were interpreted to indicate that PI-PLC apparently acts on the release of LPL in an indirect manner, releasing heparan sulphate to which LPL is bound. As there is a previously described correlation between circulating LPL and the heparin-releasable LPL, we hypothesize that the activity of PI-PLC in the endothelial cell membrane or plasma phosphatidyl-specific phospholipase D regulates the plasma LPL levels. PMID- 2554976 TI - Inhibition of aldosterone secretion by atrial natriuretic peptide in chicken adrenocortical cells. AB - Dispersed chicken adrenocortical cells were preincubated with atrial natriuretic peptide (rANP), sodium nitroprusside (SNP) or 8-bromo cyclic GMP, followed by incubations with ACTH, chicken PTH, cholera toxin or various steroid intermediates of aldosterone production. Cyclic AMP production and aldosterone secretion were evaluated, in order to determine the sites of ANP inhibition in the sequence of events leading to aldosterone secretion. Dose-dependent inhibitory effects on ACTH-stimulated aldosterone secretion by rANP and SNP were observed. Both agents appeared to stimulate cGMP production by the particulate fraction of the avian adrenocortical cells. Aldosterone production, stimulated by cyclic AMP agonists such as ACTH, chicken PTH and cholera toxin, was significantly inhibited by ANP. On the other hand, ANP did not interfere with production or degradation of cAMP. Each of the aldosterone intermediates- pregnenolone, progesterone, 11-deoxycorticosterone and corticosterone--promoted aldosterone production when included in the incubation media. Atrial natriuretic peptide and SNP inhibited aldosterone secretion when enhanced by the intermediates, by about 40-60%, but the ACTH-stimulated secretion was inhibited by over 90%. The results suggest two sites of inhibition by ANP in the pathway of aldosterone synthesis and secretion: synthesis of cholesterol or pregnenolone, and conversion of corticosterone to aldosterone. The inhibition by 8-bromo cGMP of aldosterone secretion and the similar sites of inhibition for ANP and SNP suggest that cyclic GMP mediates the inhibition in both cases. PMID- 2554977 TI - Beta-adrenergic inhibition of AGEPC-stimulated Na+/Ca2+ exchange and AGEPC induced platelet activation. AB - Recently, AGEPC (1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) was found to initiate contraction of ileal smooth muscle strips and to enhance Na+/Ca2+ exchange in ileal plasmalemmal vesicles. In the present study, the effects of the smooth muscle relaxant, isoproterenol, on Na+/Ca2+ exchange in rat ileal plasmalemmal vesicles was examined. In this preparation, Na+/Ca2+ exchange was stimulated 131 +/- 8% and 264 +/- 19% by addition of 50 nM and 100 nM AGEPC, respectively. Isoproterenol, a beta-adrenergic agonist, inhibited AGEPC stimulation of Na+/Ca2+ exchange in a dose- and time-dependent manner but had no effect on basal rates of Na+/Ca2+ antiport. At 1 microM, isoproterenol inhibited 86% of the Na+/Ca2+ exchange stimulated by 50 nM AGEPC. Vesicular cAMP levels were increased over 100% following the addition of 1 microM isoproterenol for 30 s. Inhibition of AGEPC-stimulated vesicular Na+/Ca2+ exchange and elevation of vesicular cAMP levels by isoproterenol was prevented by the beta-receptor antagonist propranolol (5 microM), demonstrating that these effects of isoproterenol were mediated by interaction with vesicular beta-adrenergic receptors. Additional studies with washed rabbit platelets demonstrated that isoproterenol inhibited AGEPC-induced aggregation and serotonin release. These effects of isoproterenol were dose- and time-dependent and were antagonized by propranolol. Isoproterenol had no effect on thrombin-induced aggregation and did not change appreciably platelet cAMP levels. Moreover, dibutyryl cAMP could not mimic the effect of isoproterenol to inhibit an AGEPC-induced aggregation. On a molar basis, the inhibitory effects of isoproterenol toward AGEPC action were greater in the ileal preparation than in the platelets. It is suggested that beta adrenergic agonists may modulate AGEPC-induced ileal Na+/Ca2+ exchange and AGEPC induced platelet aggregation through cAMP-dependent and-independent mechanisms, respectively. PMID- 2554978 TI - Nicardipine as a Ca2+ channel blocker in single barnacle muscle fibers. AB - A study has been made of the efficacy of nicardipine as a Ca2+ channel blocker by determining the magnitude of its effect on the stimulatory response of the ouabain-insensitive Na+ efflux in single barnacle muscle fibers to 100 mM external K+. The results show that nicardipine (at pH 6.5) is a potent inhibitor, the minimal effective concentration being approx. 10(-7) M and the IC(50) about 5.10(-6) M. Nicardipine, however, is not as potent as verapamil (at pH 6.5) on an equimolar basis. This is explained by assuming that the number of dihydropyridine receptors in the t-tubule membranes of barnacle fibers is not high or that verapamil is able to block the sarcoplasmic reticulum Ca2+ release channel in addition to the voltage-dependent Ca2+ channels. PMID- 2554979 TI - Characterization of fatty acid interaction with ouabain and vanadate binding to (Na+ + K+)-activated ATPase. AB - The candidateship of unsaturated fatty acids as endogenous ouabain-like factors was studied. Binding of the artificial ligand vanadate at the intracellular phosphorylation epitope of membrane-bound Na+/K+-ATPase was unaffected by linoleic and arachidonic acid. In the (Mg2+ + Pi)-facilitated system for ouabain binding they were characterized as noncompetitive inhibitors of cardiac glycoside binding, however. The ouabain binding capacity as well as the affinity decreased and the ouabain dissociation rate was accelerated by fatty acids. In the presence of vanadate for facilitation of ouabain binding an increase in ouabain affinity was seen. It is concluded that elementary criteria for the characterization of unsaturated fatty acids as ouabain-like factors are not fulfilled. The ratio between E2-subconformations of Na+/K+-ATPase with different ouabain affinities may be changed by incorporation of fatty acids in the lipid membrane. PMID- 2554980 TI - The orientation of (-)-delta 9-tetrahydrocannabinol in DPPC bilayers as determined from solid-state 2H-NMR. AB - The orientation of the motional axis of (-)-delta 9-tetrahydrocannabinol in dipalmitoylphosphatidylcholine model membrane was calculated from the 2H quadrupolar splittings (delta nu Q) of individual deuterons strategically located on the cannabinoid tricyclic component. The molecule assumes an orientation in which its long axis is nearly perpendicular to the phospholipid chains and its most ordered axis is almost in the plane of the aromatic ring. This 'awkward' cannabinoid orientation in the membrane presumably occurs in order to allow the phenolic hydroxyl group to direct itself towards the polar bilayer interface. PMID- 2554981 TI - Location and dynamics of alamethicin in unilamellar vesicles and thylakoids as model systems. A spin label study. AB - Location and dynamics of the voltage-dependent pore-forming icosapeptide alamethicin have been studied using spin labels which were linked directly and via spacers to the C-terminus of the amphiphilic alpha-helix. Ion-transport activities of these derivatives were found to be very similar to those of natural alamethicin in green plant thylakoids chosen as a model system. The shape of the electron spin resonance spectra indicates segmental motion of the nitroxide rather than rotation of the whole peptide. A population of spins showing narrow lines in the presence of thylakoids or lipid vesicles is attributed to alamethicin in the aqueous solution. A second population shows rotational correlation times greater than 10(-9) s and is bound to the membranes, the C termini residing in an environment with a polarity close to that of water. This population is inaccessible to the hydrophilic, charged line broadening agent chromium oxalate. Since spectral shapes and amplitudes of spectra are unchanged by additions of unlabelled peptide, it is concluded that the ESR detectable spins are bound to peptides essentially in the monomeric state. Alamethicin induced pore formation under flash illumination is demonstrated by measurement of kinetics of proton deposition in the thylakoid interior. When pores are opened by illuminating thylakoids and thus applying a membrane potential, mainly the bound population is affected by a process reversibly suppressing the signal, whereas only limited disappearance of label from the external medium is detected. Apparently, the potential causes a change in the conformation of the peptide which leads to a further immobilisation of the label, possibly due to a deeper insertion of the alpha-helices into the lipid membrane. However, evidence has been presented experimentally that there is no detectable change of potential prior to the opening of the pore. PMID- 2554982 TI - An electron paramagnetic resonance study of skeletal muscle membrane fluidity in malignant hyperthermia. AB - Skeletal muscle sarcolemma (SL), transverse tubule (TT) and heavy sarcoplasmic reticulum (HSR) membranes were isolated from malignant hyperthermia susceptible (MHS) and normal pigs, and the rotational dynamics of lipid hydrocarbon chain motion was examined by electron paramagnetic resonance (EPR) spectroscopy. The stearic acid spin probe 16-SASL was incorporated into MHS and normal membranes and both the order parameter (S) and effective correlation time (tau r) of probe motion were calculated from spectra recorded over the temperature range of 2 to 40 degrees C. At any given temperature, TT membranes exhibited significantly greater values for both the S and tau r of probe motion than did SL, which exhibited significantly greater values than did HSR membranes. The order of decreasing S and tau r values for 16-SASL mobility correlated with the decreasing cholesterol content of these membranes (TT greater than SL greater than HSR), however there was no difference in the S or tau r values for a given membrane fraction isolated from both MHS and normal muscle. Arrhenius plots of 16-SASL mobility in SL, TT and HSR were linear from 2 to 40 degrees C, indicating no abrupt thermotropic change in the lipid hydrocarbon phase of any of the membrane types studied. Apparent activation energies (Ea), calculated from the Arrhenius plots, were similar for MHS and normal membranes derived from a given cellular location. However, the Ea of probe motion for TT membranes (2.3 +/- 0.1 and 2.4 +/- 0.1 kcal/mol/degree for MHS and normal, respectively) was significantly less than for SL (3.4 +/- 0.4 and 2.9 +/- 0.1 kcal/mol/degree for MHS and normal, respectively) which, in turn, was significantly less than the Ea for HSR (3.7 +/- 0.1 and 3.7 +/- 0.1 kcal/mol/degree for MHS and normal, respectively). Since 16 SASL motion was similar in MHS and normal membranes, we conclude that there is no evidence for a generalized membrane defect affecting lipid mobility in these MHS muscle membranes. PMID- 2554983 TI - cAMP-dependent protein kinase phosphorylates gap junction protein in lens cortex but not in lens nucleus. AB - MP70 (a 70 kDa membrane protein) is a component of the gap junctions of the young fibre cells in the lens outer cortex. In the older fibres deeper in the mammalian lens (lens nucleus), MP70 is processed to MP38 by cleavage and removal of the carboxy terminal half. It is shown here that cortical MP70, and its derivative MP64, can be phosphorylated with cAMP-dependent protein kinase. In contrast, MP38 from the lens nucleus is not phosphorylated by the enzyme. Proteolytic processing and this lens region specific phosphorylation are relevant for the future development of functional assays for lens gap junctions. PMID- 2554984 TI - Effects of L-carnitine and palmitoylcarnitine on membrane fluidity of human erythrocytes. AB - Amphiphilic compounds such as long-chain acyl carnitine accumulate in ischemic myocardium and potentially contribute to the myocardial damage, and the role of carnitine in protecting the heart against ischemic damage is interesting. It has been reported that palmitoylcarnitine causes alterations in the membrane molecular dynamics, so this study was designed to investigate whether L-carnitine had a stabilizing effect of membrane fluidity using the spin-label technique. Human erythrocytes were spin-labeled with 5-doxylstearic acids, and membrane fluidity was quantified by measuring the change in the order parameter S. The administration of palmitoylcarnitine (100 microM) altered the membrane fluidity of erythrocytes and caused significant morphological changes. L-carnitine (2mM) decreased the alteration of the fluidity of erythrocytes incubated with palmitoylcarnitine (100 microM), and improved the morphological changes in erythrocytes. These results show that L-carnitine has a stabilizing effect of membrane fluidity as a result of interaction with the palmitoylcarnitine which has a detergent effect. PMID- 2554985 TI - [Purification of phospholipase C isoenzyme 1 from Clostridium perfringens and its properties]. AB - A procedure for the purification of isoenzyme I of phospholipase C from Cl. perfringens was developed. The isoenzyme was purified to homogeneity (data from disc electrophoresis) using affinity chromatography on polycephamide and gel filtration through Ultrogel AcA-54, the enzyme yield being 41%. Some properties of the purified isoenzyme I (pH and temperature optima, stability, effect of metal ions and detergents, substrate dependence) were investigated. No significant differences between the properties of the unfractionated enzyme and isoenzyme I were established. PMID- 2554986 TI - [Regulation by Ca2+-calmodulin-dependent phosphorylation of passive transport of Ca2+ in the myometrial sarcolemma]. AB - Closed vesiculate preparations of pig myometrium sarcolemma (predominantly with inside-out orientation) are characterized by passive permeability for Ca2+. The kinetics of Ca2+ release from the vesicles is exponential. Using the grapho analytical subtraction method, the kinetic parameters of this reaction were determined. Myometrium sarcolemma contains endogenous Ca2+-calmodulin-dependent protein kinase and phosphoprotein phosphatase which is inhibited by sodium o vanadate. The Ca2+-calmodulin-dependent phosphorylation stimulates passive Ca2+ release from sarcolemmal vesicles. In the course of phosphorylation the capacity of the pool providing for rapid Ca2+ release increases by 61%, the initial rate of Ca2+ release showing a 28% increase. Trifluoroperazine, an inhibitor of Ca2+ calmodulin-dependent processes, eliminates the activating effect of phosphorylation on the rate of Ca2+ release from sarcolemmal vesicles. PMID- 2554987 TI - [Effect of inhomogenous tissue on noninvasive detection of the velocity of evoked transmission in motor muscle units--study with the trough model]. PMID- 2554988 TI - Interaction of water with oriented DNA in the A- and B-form conformations. AB - High resolution 2H nuclear magnetic resonance (NMR) was used to investigate the interaction of D2O with solid samples of uniaxially oriented Li-DNA (B-form DNA) and Na-DNA (A- and B-form DNA). At low levels of hydration, 0 approximately 4 D2O/nucleotide, the 2H spectra shows a very weak (due to short T2) broad single resonance, suggestive of unrestricted rotational diffusion of the water. At approximately 5 or more D2O/nucleotide, the Li-DNA (B-form) spectra suddenly exhibit a large doublet splitting, characteristic of partially ordered water. With increasing hydration, the general trend is a decrease of this splitting. From our analysis we show that the DNA water structure reorganizes as the DNA is progressively hydrated. The D2O interaction with Na-DNA is rather different than with Li-DNA. Below 10 D2O/nucleotide Na-DNA is normally expected to be in the A form, and a small, or negligible splitting is observed. In the range 9-19 D2O/nucleotide, the splitting increases with increasing hydration. Above approximately 20 D2O/nucleotide Na-DNA converts entirely to the B-form and the D2O splittings are then similar to those found in Li-DNA. We show that the complex Na-DNA results obtained in the range 0-20 D2O/nucleotide are caused by a mixture of A- and B-DNA in those samples. PMID- 2554989 TI - Low-frequency collective modes in dry and hydrated proteins. AB - We have observed Brillouin-like low frequency collective modes in the scattering of 1 A neutrons from a fully in vivo deuterated protein. These modes are tentatively interpreted as due to short-lived coherent excitations propagating with velocities between 2,000 and 4,000 m/s in elements of the secondary structure and patches of closely associated water. PMID- 2554991 TI - Activation of calcium channels in sarcoplasmic reticulum from frog muscle by nanomolar concentrations of ryanodine. AB - Sarcoplasmic reticulum vesicles isolated from fast-twitch frog skeletal muscle presented two classes of binding sites for ryanodine, one of high affinity (Kd1 = 1.7 nM, Bmax1 = 3.3 pmol per mg) and a second class with lower affinity (Kd2 = 90 nM, Bmax2 = 7.0 pmol per milligram). The calcium channels present in the sarcoplasmic reticulum membranes were studied in vesicles fused into lipid bilayers. Low concentrations of ryanodine (5 to 10 nM) activated a large conductance calcium channel after a short delay (5 to 10 min). The activation, which could be elicited from conditions of high or low fractional open time, was characterized by an increase in channel fractional open time without a change in conductance. The open and closed dwell time distributions were fitted with the sum of two exponentials in the range of 4 to 800 ms. The activating effect of ryanodine was due to an increase of both open time constants and a concomitant decrease in the closed time constants. Under conditions of low fractional open time (less than 0.1), the time spent in long closed periods (greater than 800 ms) between bursts was not affected by ryanodine. Higher concentrations of ryanodine (250 nM) locked the channel in a lower conductance level (approximately 40%) with a fractional open time near unity. These results suggest that the activating effects of nanomolar concentrations of ryanodine may arise from drug binding to high affinity sites. The expression of the lower conductance state obtained with higher concentrations of ryanodine may be associated with the low affinity binding sites observed in frog sarcoplasmic reticulum. PMID- 2554993 TI - Identification through chemical cross-linking of distinct granulocyte-macrophage colony-stimulating factor and interleukin-3 receptors on myeloid leukemic cells, KG-1. AB - Granulocyte/macrophage colony-stimulating factor (GM-CSF) and interleukin-3 (IL 3) each bind specifically to a small number of high-affinity receptors present on the surface of the cells of the acute myelogenous leukemia line, KG-1. Through chemical cross-linking of IL-3 and GM-CSF to KG-1 cells, we identified distinct binding proteins for each of these cytokines with approximate molecular masses of 69 and 93 Kd, respectively. Although these two binding proteins are distinct, GM CSF and IL-3 compete with each other for binding to KG-1 cells. Other cell lines, which express receptors for either factor but not for both do not display this cross-competition for binding with IL-3 and GM-CSF. These findings imply that distinct IL-3 and GM-CSF binding proteins are expressed on the cell surface and that an association exists between these proteins on KG-1 cells. PMID- 2554990 TI - Selective detection of the rotational dynamics of the protein-associated lipid hydrocarbon chains in sarcoplasmic reticulum membranes. AB - We have developed a saturation transfer EPR (ST-EPR) method to measure selectively the rotational dynamics of those lipids that are motionally restricted by integral membrane proteins and have applied this methodology to measure lipid-protein interactions in native sarcoplasmic reticulum (SR) membranes. This analysis involves the measurement of spectral saturation using a series of six stearic acid spin labels that are labeled with a nitroxide at different carbon atom positions. A large amount of spectral saturation is observed for spin labels in native SR membranes, but not for spin labels in dispersions of extracted SR lipids, implying that the motional properties of those lipids interacting with the Ca-ATPase, i.e., the boundary or annular lipid, can be directly measured without the need for spectral subtraction procedures. A comparison of the motional properties of the restricted lipid, measured by ST EPR, with those measured by digital subtraction of conventional EPR spectra qualitatively agree, for in both cases the Ca-ATPase restricts the rotational mobility of a population of lipids, whose rotational mobility increases as the nitroxide is positioned toward the center of the bilayer. However, the ability of ST-EPR to directly measure the motionally restricted lipid in a model-independent means provides the greater precision necessary to measure small changes in the rotational dynamics of the lipid at the protein-lipid interface, providing a valuable tool in clarifying the relationship between the physical nature of the protein-lipid interface and membrane function. PMID- 2554992 TI - Rate of allosteric change in hemoglobin measured by modulated excitation using fluorescence detection. AB - We have measured the forward and reverse rates of the allosteric transition of hemoglobin A with three CO molecules bound by using modulated excitation coupled with fluorescence quenching of the DPG analogue, PTS (8-hydroxy-1,3,6 pyrene trisulfonic acid). This dye is observed to bind to the T state with significantly larger affinity than to the R state, and thus provides an unequivocal marker for the molecule's conformational change. The allosteric rates obtained with the fluorescent dye (pH 7.0, bis-Tris buffer) are (3.4 +/- 1.0) x 10(3)s-1 for the R to T transition and (2.1 +/- 0.5) x 10(4)s-1 for the T to R transition. This gives an equilibrium constant L3 of 0.16 +/- 0.06. These results provide good agreement with modulated difference spectra calibrated from model compounds, arguing that there is little if any difference in the kinetics observed by the heme spectra and the kinetics of the full subunit motion. The equilibrium constant between structures (L3) is smaller in the absence of phosphates than observed in phosphate buffer (0.33). However, the rates of the allosteric transition increase in the absence of phosphates as compared with the corresponding rates in phosphate buffer of 1.0 x 10(3)s-1 and 3.0 x 10(3)s-1. The effects of inorganic phosphates on the equilibrium can be separated from the effects on kinetics. We find that phosphates also affect the dynamic behavior of hemoglobin, and the presence of 0.15 M phosphate can be viewed as raising the transition state energy between R and T conformations by approximately 0.5 kcal/mol exclusive of the T state stabilization. Dissociation constants for the dye were measured to be 104 +/- 25 microM for unligated T state and 930 +/- 300 microM for the fully ligated R state. The best fit equilibrium constant (125 +/- 40 microM) for three ligands bound does not differ significantly from that measured without ligands bound. Incidental to the measurement technique is the determination of the rates of binding and release of the dye. The association rate for binding to the T state is large, (at least 4 x 10(9) M-1 s-1) and may be diffusion limited, while the association and dissociation rates for R state binding, while not determined with precision, are clearly much smaller, of the scale of 10(5) M-1 s-1 for association. PMID- 2554994 TI - Rearrangements of T-cell antigen receptor delta chain gene in hematologic neoplasms. AB - Rearrangements of the T-cell antigen receptor (TCR) delta chain gene were studied in primary neoplastic cells from 137 patients with leukemia or lymphoma. TCR delta gene rearrangements or deletions were observed in all 50 T-cell neoplasms: 5 of 8 CD3- T-cell neoplasms showed rearrangements, whereas biallelic deletion of TCR delta gene was the most common pattern in CD3+ T-cell neoplasm (39 of 42 patients). Rearrangements of TCR delta gene were also detected in 23 of 40 immature B-cell leukemias, including 22 of 25 patients with rearrangements of TCR gamma gene, 2 of 17 mature B-cell neoplasms, and 3 of 30 myeloid leukemias. Thus, TCR delta gene rearrangement or deletion is always found in T-cell neoplasms and is frequently found in immature B-cell leukemias associated with TCR gamma gene rearrangement. Furthermore, TCR delta gene rearrangements associated with the germline configuration of the TCR beta, gamma, and immunoglobulin heavy chain genes were observed in two immature T-cell leukemias, suggesting that TCR delta gene rearrangements precede TCR gamma and beta gene rearrangements. These results indicate that an analysis of TCR delta gene rearrangement provides potential tools to establish the clonality of immature T-cell neoplasms and to identify the normal stages of lymphocyte differentiation. PMID- 2554996 TI - Biotinylated erythropoietin. PMID- 2554995 TI - SUP-HD1: a new Hodgkin's disease-derived cell line with lymphoid features produces interferon-gamma. AB - A new cell line, SUP-HD1, was established from the pleural effusion of a patient with nodular sclerosing Hodgkin's disease (NSHD). The SUP-HD1 cells had the characteristic morphology of Reed-Sternberg cells and contained acid phosphatase and nonspecific esterase. The cells lacked the Epstein-Barr virus (EBV) genome and reacted with monoclonal antibodies (MoAbs) against CD15 (Leu-M1), CD25 (Tac), CD71 (OKT9), Ki67, and HLA-Dr. However, the SUP-HD1 cells were nonreactive with MoAbs that specifically identify T lymphocytes, B lymphocytes, and macrophage/myeloid cells. Karyotype analysis of the cell line showed clonal abnormalities involving 1p13, 7p15, 8q22, and 11q23, chromosomal locations, at which breakpoints have been reported in HD. Southern blot analysis demonstrated rearrangement of the immunoglobulin heavy chain and kappa light chain genes as well as the gene for the beta chain of the T-cell receptor (TCR). Transcriptional analysis showed expression of RNAs for kappa light chain, interferon-gamma (IFN gamma), and interleukin-2 receptor (IL-2R) but not IL-2. The SUP-HD1 cells lacked cytoplasmic and surface immunoglobulin heavy chain, but a small amount of cytoplasmic kappa light chain was detected. The presence of nuclear factor kappa B (NF kappa B), a B-lymphocyte-associated transcription factor, was demonstrated in stimulated and unstimulated cells. In addition, the SUP-HD1 cell line, produced IFN-gamma, a T-lymphocyte-associated lymphokine. Based on these data, the SUP-HD1 cells appear to be aberrant lymphocytes with characteristics of both activated B and T lymphocytes. Elaboration of lymphokines such as IFN-gamma by the malignant cells may represent one explanation for the unique clinical and pathologic features of HD. PMID- 2554997 TI - Segmental neogenesis of the dog esophagus utilizing a biodegradable polymer framework. AB - This study evaluated the ability of biodegradable implants fabricated from polymers and co-polymers of polylactic acid (PLA) and polyglycolic acid (PGA) to induce regeneration of surgically created defects in the dog esophagus. The study utilized 12 mongrel dogs that had a 5 cm segment of the esophagus removed. Implants were fabricated by spray casting the polymers on a spinning Teflon mandril. The defects were repaired by suturing the biodegradable implants to the proximal and distal ends of the esophagus. Ten of the dogs were sacrificed from 3 days to 8 weeks after surgery while 1 of the dogs died after 3 years and 1 dog was sacrificed 4 years after graft placement. Endoscopic and histologic examination of the grafts 3 days after placement showed minimal inflammatory response and an apparent seal between the esophagus and implant at the suture lines. Two weeks after surgery a fibrous connective tissue sheath, continuous with the proximal and distal segments of the esophagus, could be seen surrounding the graft. One month after placement, the implants were partially degraded leaving a connective tissue repair continuous with the proximal and distal ends of the esophagus. The repair area was lined with epithelium and enabled the dogs to drink freely and eat semisolid foods. In conclusion, it has been shown that it is possible to fabricate a biodegradable implant which can stimulate regeneration of a hollow organ and which is compatible with long term survival. PMID- 2554998 TI - Comparative evaluation of the structure and properties of certain granulated and fibrous activated carboniferous sorbents. AB - This paper presents the results of the comparative evaluation of the structure and sorption properties of fibrous (AYBM- -MH) and granulated (CKH-IK) carboniferous sorbents. The structure of sorbents was investigated using scanning electron microscopy (SEM) and electron paramagnetic resonance (EPR), while the sorption properties were studied through application of adsorption-desorption of markers (medinal, creatinine, uric acid, vitamin B12). Carbon fiber sorbents have been found to possess more favorable characteristics of the sorptive capacity and a better porous structure. The EPR spectrum of a fibrous sorbent contains the signal in the form of a solitary line with a g-factor of 2.0025 +/- 0.0003, while the granulated sorbent has no signal. Such properties of fibrous sorbents as nontoxicity, biological compatibility, variety of textile patterns, elasticity, strength, etc. promote their application in capacity of versatile, high-quality medical materials. Due to their compatibility with blood cells and body tissue, spherical sorbents found extensive application in the clinical practice (1,2). However, considering the potential application of sorbents in modern medicine (hemosorption, enterosorption, application therapy) it is important that sorbents with improved sorption, kinetic and other operational characteristics be developed. The results presented demonstrate properties of granulated and fibrous sorbents produced using different technologies and source materials. Also indicated are the differences related to the pattern of sorbents. PMID- 2554999 TI - [Retinoid receptors in epidermoid carcinoma of the ORL sphere]. AB - Head and neck tumors from 20 patients were investigated for the presence and the level of retinoid binding proteins (cellular retinoic acid binding protein or CRABP, cellular retinol binding protein or CRBP). A binding assay-gel filtration technic was used. CRABP were found in all tumors and in the adjacent normal tissues. A decreased level was associated with poor prognostic factors. CRBP were only found in some tumors and were absent from the adjacent normal tissue and from benign tumors. These results are discussed as regards their interest in prognostic evaluation and therapeutic orientation. PMID- 2555000 TI - Overtreatment of children with Wilms' tumour outside paediatric oncology centres. PMID- 2555001 TI - Persistence of cytomegalovirus in mononuclear cells in peripheral blood from blood donors. PMID- 2555002 TI - Antenatal prophylaxis with anti-D immunoglobulin. PMID- 2555003 TI - Analytic results for quantifying HIV infectivity. AB - In a separate paper, we developed a mathematical model describing HIV infection and used it to suggest experiments for quantifying characteristic viral parameters. In this paper we generalize the model to any well-mixed assay system. We also present complete and rigorous derivations of fundamental results needed for the design and analysis of HIV infectivity assays. The model is applicable to infectious agents with multiple receptors for their target cell (e.g. HIV, Epstein-Barr virus and Plasmodium), and to blockers (both reversible and irreversible), as long as blocker and target cells are the same diffusion compartment. PMID- 2555004 TI - Variables predicting deep fungal infections in bone marrow transplant recipients. AB - Deep fungal infections (FI) were diagnosed in 27 out of 209 consecutive bone marrow transplantation (BMT) recipients. Autopsy verified that the incidence of deep FI was 10% and the overall incidence was 13%. Using bivariate logistic regression analysis at the time of BMT, high recipient age (p = 0.003), low bone marrow cell dose (p = 0.007), recipient cytomegalovirus (CMV) seropositivity (p = 0.009) and splenectomy (p = 0.03) were significant risk factors for deep FI. In multivariate analysis, splenectomy (p = 0.008), recipient CMV seropositivity (p = 0.01) and low bone marrow cell dose (p = 0.01) held as significant. At 30 days post-BMT anti-thymocyte globulin treatment (p = 0.0006) and graft-versus-host disease grade II-IV (p = 0.005) were significant risk variables in bivariate logistic regression analysis and Fisher's exact probability test. Patients with these risk factors are candidates for treatment with antifungal drugs when they suffer from leukopenia and unclear fever. PMID- 2555005 TI - Pharmacokinetics of cytomegalovirus specific IgG antibody following intravenous immunoglobulin in bone marrow transplant patients. AB - Cytomegalovirus (CMV) associated interstitial pneumonitis is a major cause of death among bone marrow transplant patients. A variety of intravenous immunoglobulin (IVIg) preparations have shown some promise in preventing this complication. As part of a multicenter trial of Sandoglobulin, the pharmacokinetics of CMV specific IgG was measured in order to guide future dosing schedules. A dose of 500 mg/kg was administered weekly beginning 1 week before transplant and continuing until day 98 following transplant. The half-life of CMV specific IgG was measured by an ELISA method after the first, third and fifth doses of IVIg. CMV seronegative patients received only screened CMV negative blood products, which permitted assessment of the half-life of IVIg CMV antibody. Peak titers achieved were comparable to those of the CMV seropositive patients averaging 1:2702 (range 1:596-1:10 514). Total IgG levels rose to a peak of about 75% above baseline. After the first dose of IVIg, the half-life of CMV IgG antibody was 3.4 +/- 2.0 (SD) days, although it lengthened to 6.1 +/- 5.1 days after the fifth dose of IVIg. The half-life of total IgG was estimated to be between 5 and 10 days, depending on the assumptions made regarding endogenous production. If high levels of IVIg are necessary for protection from CMV associated interstitial pneumonitis, weekly dosing will be important in order to maintain sufficient levels to be protective. PMID- 2555006 TI - Foscarnet prophylaxis in marrow transplant recipients. PMID- 2555007 TI - Angiotensin converting enzyme activity is present in the endothelium-denuded aorta. AB - Angiotensin converting enzyme activity was measured in intact aortic rings utilizing the synthetic tripeptide [3H]-benzoyl-Phe-Ala-Pro as the substrate. Intact aortic rings possessed angiotensin converting enzyme activity which was blocked by captopril. Enzyme activity was reduced by approximately 30% after removal of the endothelium by chemical or mechanical methods. The remaining activity was also captopril-sensitive. These results suggest that in addition to endothelium, angiotensin converting enzyme activity is present in other vascular cells and may contribute to the metabolism of angiotensin I generated within the vessel wall. PMID- 2555008 TI - The effect of M&B 22948 on methacholine- and histamine-induced contraction and inositol 1,4,5-trisphosphate levels in guinea-pig tracheal tissue. AB - The effect of a cyclic GMP phosphodiesterase inhibitor, M&B 22948, on methacholine- and histamine-induced contraction and inositol 1,4,5-trisphosphate (IP3) elevation was studied in guinea-pig tracheal rings. After addition of methacholine or histamine the rise in IP3 content was rapid and transient reaching a maximum after 5-15 s, which coincided with the maximum rate of tension development. Cyclic GMP levels of the tissue were elevated by M&B 22948 before agonist stimulation and further elevated by addition of methacholine or histamine. Cyclic AMP levels were not altered by any of these agents. M&B 22948 abolished IP3 generation induced by methacholine or histamine, but did not alter the rate or magnitude of tension development. Thus, IP3 generation does not appear to be responsible for the contractions induced by methacholine or histamine in this tissue. PMID- 2555009 TI - An examination of the putative sigma-receptor in the mouse isolated vas deferens. AB - 1. The effects of several ligands which interact with the sigma-binding site were studied on the electrically-evoked (0.1 Hz) neurogenic twitch contractions of the mouse isolated vas deferens. 2. (+)-3-(3-Hydroxyphenyl)-N-(1-propyl)piperidine [+)-3-PPP) (10(-8) - 10(-5) M), inhibited the neurogenic twitch contractions. This inhibitory action was unaffected by naloxone (10(-6)M), idazoxan (10(-6)M), cocaine (10(-6)M) or tyramine (10(-4)-3 x 10(-4)M), but was abolished by the dopamine D2-antagonist, sulpiride (10(-6)M). Therefore, in order to study the potentiating actions of sigma ligands, sulpiride (10(-6)M) was used to prevent any inhibitory actions mediated via dopamine D2-receptors. 3. In the presence of sulpiride (10(-6)M), haloperidol (10(-6)-10(-5)M), (+)-3-PPP (10(-6)-3 x 10(-4) M) and (+)-N-allyl-N-normetazocine [+)-SKF 10,047) (10(-5)-10(-4)M) each reversibly potentiated the neurogenic twitch contractions in a concentration dependent manner. The rank order of potency was haloperidol greater than (+)-3 PPP greater than (+)-SKF 10,047. 4. The stereoisomers of 3-PPP displayed stereoselectivity with (+)-3-PPP being more potent than (-)-3-PPP. 5. At a concentration that did not potentiate the twitch contractions, (3 x 10(-7)M), haloperidol did not antagonize the potentiating action of (+)-3-PPP (3 x 10( 5)M). 6. 1,3-Di-O-tolyguanidine (DTG) (10(-8)-10(-5)M) had no effect on the amplitude of twitch contractions and did not affect the potentiating action of (+)-3-PPP (10(-5)-3 x 10(-5)M). 7. It is concluded that a-ligands potentiate neurogenic twitch contractions of the mouse isolated vas deferens via a site that is different from the central sigma-binding site. PMID- 2555010 TI - Caffeine-induced inhibition of calcium channel current in cultured smooth cells from pregnant rat myometrium. AB - 1. The inhibitory effect of caffeine on the calcium channel current was investigated in cultured myometrial cells isolated from pregnant rats. 2. Caffeine inhibited the calcium channel current elicited from a holding potential of -70 mV in a concentration-dependent manner. The IC50 was estimated to be 35 mM. 3. The caffeine inhibition was not enhanced when calcium channels were opened by a conditioning depolarizing pulse sequence or when the number of inactivated calcium channels was increased at depolarized holding potentials. 4. Caffeine antagonized the specific binding of (+)-[3H]-isradipine to myometrial membranes. The IC50 value found in binding experiments was similar to the IC50 value for half-maximal inhibition of calcium channel current. Caffeine decreased the maximal binding capacity of (+)-[3H]-isradipine to myometrial membranes without any significant change in the dissociation constant. 5. The results indicate that caffeine interacts with a site closely associated with the voltage-dependent calcium channels in myometrial cells and, in turn, inhibits calcium influx. PMID- 2555011 TI - Spinal antinociceptive actions of mu- and kappa-opioids: the importance of stimulus intensity in determining 'selectivity' between reflexes to different modalities of noxious stimulus. AB - 1. In electrophysiological experiments in spinalized rats, mu- and kappa-opioids were tested intravenously on the responses of single motoneurones to electronically controlled, alternating noxious heat and noxious pinch stimuli. The effects of mu- and kappa-opioids were compared with those of the general anaesthetic alpha-chloralose and the dissociative anaesthetic/PCP ligand ketamine. 2. The kappa-opioids U-50,488 (0.5-16 mgkg-1 i.v.) and tifluadom (0.05 1.6 mgkg-1 i.v.) had very similar actions to the mu-opioid fentanyl (0.5-16 micrograms kg-1 i.v.). Thus all three agonists reduced thermal and mechanical nociceptive reflexes in parallel and in a dose-dependent manner, but only so long as neuronal responses to the alternating stimuli elicited similar excitability levels in the neurone under study. Ketamine (0.5-16 mgkg-1 i.v.) had similar actions to the opioids whereas alpha-chloralose (20 mgkg-1 i.v.) had very little effect on neuronal responsiveness. 3. Apparently 'selective' depressions by both mu- and kappa-opioids could be orchestrated by a deliberate mismatch of the intensities of alternating noxious heat and pinch stimuli; as measured by neuronal firing rate, the weaker of the responses to either type of stimulus was invariably reduced to a greater degree. 4. Similar 'selectivity' could be demonstrated for both mu- and kappa-ligands when the weaker and stronger responses were of the same modality, being applied by the same pincher device but with alternating applied force. 5. It is concluded that the 'selective' spinal actions of kappa-opioids seen in non-thermal over thermal behavioural models of nociception is likely to be related to the relative intensities, rather than the modalities, of the noxious stimuli used. The validity of the interpretation of results obtained in such behavioural studies is discussed. PMID- 2555012 TI - Spinal antinociceptive actions and naloxone reversibility of intravenous mu- and kappa-opioids in spinalized rats: potency mismatch with values reported for spinal administration. AB - 1. The relative spinal effectiveness of mu- and kappa-opioids has been assessed by their intravenous potencies on nociceptive responses (heat and/or pinch) of single motoneurones recorded in alpha-chloralose anaesthetized, spinalized rats. 2. The depressant actions of both mu- and kappa-opioids were reversed by low intravenous doses of naloxone (10 to 100 micrograms kg-1). When tested at a dose of 1 microgram kg-1 i.v., naloxone antagonized the effects of the mu-agonist morphine but had no effect on the kappa-opioid U-50,488. This provides further support for the theory that the actions of mu- and kappa-ligands were mediated at different subclasses of opioid receptor but highlights the difficulties in using antagonists with poor receptor selectivity to differentiate between mu- and kappa receptor-mediated effects in vivo. 3. The molar potency rations of fentanyl: morphine:U-50,488: tifluadom for thermal and mechanical nociceptive responses were 620: 1.0:0.74:5.7 and 520:1.0:0.56:7.7 respectively. These potency ratios, as well as the absolute potencies, agree well with those reported in several behavioural studies in which systemic administration of agonists was used in non thermal tests. 4. The agonist potency values obtained in this study contrast with those reported for local spinal administration. By this route, the potency of lipophilic opioids (e.g. fentanyl, U-50,488 and tifluadom) relative to hydrophilic opioids (e.g. morphine) is much reduced, implying that activity of intrathecally administered opioids is more dependent on the physico-chemical properties of the agonists used than on the relative abundance in the spinal cord of functional opioid receptors of the mu- and kappa-subtypes. This conclusion indicates that the results with locally applied opioids should not be used to assess spinal opioid receptor function. PMID- 2555013 TI - On the selectivity of intravenous mu- and kappa-opioids between nociceptive and non-nociceptive reflexes in the spinalized rat. AB - 1. In electrophysiological experiments in spinalized, alpha-chloralose anaesthetized rats, opioids and anaesthetics were tested intravenously (i.v.) on the responses of individual motoneurones to alternating noxious (heat or pinch) and non-noxious (tap or vibration) stimuli. 2. On cells that were sensitive to low doses of mu-opioids, both fentanyl (0.5-4 micrograms kg-1 i.v.) and morphine (0.5 mg kg-1 i.v.) selectivity reduced reflexes to noxious stimuli to a greater degree than the higher doses required to reduce nociceptive reflexes (fentanyl 8 micrograms kg-1 i.v.; morphine 1-8 mg kg-1 i.v.) depressed non-nociceptive reflexes to a similar degree. 3. A similar spectrum of selectivity was seen with U-50,488 (0.5-16 mg kg-1 i.v.) although statistically significant selective depression of reflexes was only evident at the lowest dose tested (0.5 mg kg-1 i.v.). All effects of U-50,488 were readily reversed by low doses of the opioid antagonist, naloxone (10-100 micrograms kg-1 i.v.). 4. The dissociative anaesthetic/PCP ligand ketamine (0.5-4 mg kg-1 i.v.) was similar in having selective actions at low doses on sensitive cells but non-selective actions when higher doses were required. In contrast, the general anaesthetics methohexitone (4 mg kg-1 i.v.) and alphadolone/alphaxalone (1 mg kg-1 i.v.) were consistently non-selective between reflexes to noxious and non-noxious stimuli. alpha Chloralose (20-40 mg kg-1 i.v.) had very little effect on reflexes to any of the synaptic inputs tested. PMID- 2555014 TI - Inhibition of agonist-stimulated inositol lipid metabolism by the anticonvulsant carbamazepine in rat hippocampus. AB - 1. The effect of the anticonvulsant, anti-manic drug carbamazepine was examined on inositol lipid signalling in rat hippocampus in vitro. 2. Hippocampal miniprisms were labelled with [3H]-inositol before stimulation with a variety of neuroactive agents that increase phosphoinositide turnover. 3. The presence of carbamazepine (0.1-100 microM) during labelling caused a dose-related reduction of basal and carbachol-evoked [3H]-inositol phosphate accumulations. The effect of the drug on basal inositol phosphate levels was lost when slices were labelled with [3H]-inositol before incubation with carbamazepine. 4. Incubation of slices with carbamazepine after labelling with [3H]-inositol and before stimulation showed the inhibitory effect of the drug to be selective according to the agonist used. Responses to carbachol, histamine and the sodium-channel agent veratrin were reduced by carbamazepine whilst the responses to 5-hydroxytryptamine, noradrenaline and substance P were unaffected. 5. Inhibition of carbachol, histamine and veratrin-induced stimulation by carbamazepine share a similar dependence on length of pre-incubation time with the drug. However, the effect of carbamazepine (100 microM) on the respective dose-response curves suggests that the mechanism of inhibition of the carbachol response differs from the inhibition of the histamine and veratrin responses. These effects may be significant in the mechanism of action of carbamazepine as an anticonvulsant and in its effectiveness against manic depression. PMID- 2555015 TI - Inhibitory effect of a toxin okadaic acid, isolated from the black sponge on smooth muscle and platelets. AB - 1. Effects of okadaic acid, a toxin isolated from marine sponges, on smooth muscle contraction and platelet activation were examined. 2. Contractions in rabbit aorta induced by high concentrations of K+ and noradrenaline were inhibited by 0.1-1 microM okadaic acid in a concentration-dependent manner. Spontaneous rhythmic contractions as well as high K+-induced contraction in guinea-pig taenia caeci were also inhibited by 1 microM okadaic acid. 3. High K+ induced contraction in rabbit aorta was accompanied by increased Ca2+ influx measured with 45Ca2+ and increased cytosolic Ca2+ [( Ca2+]cyt) measured with fura 2-Ca2+ fluorescence. Okadaic acid inhibited the contraction without inhibiting Ca2+ influx and produced only a small decrease in [Ca2+]cyt. 4. In a saponin skinned taenia, Ca2+-induced contraction was not inhibited but rather potentiated by okadaic acid. 5. Okadaic acid, 1 microM, inhibited aggregation, ATP release and increased in [Ca2+]cyt induced by thrombin in washed rabbit platelets. Okadaic acid itself did not change the platelet activities. 6. Okadaic acid did not change the cyclic AMP content of rabbit aorta although the inhibitory effects of okadaic acid were similar to those of cyclic AMP. 7. Although the mechanism of the inhibitory effect of okadaic acid was not clarified in the present experiments, it is suggested that okadaic acid acts by inhibiting protein phosphatases resulting in an indirect activation of cyclic AMP-dependent protein phosphorylation. PMID- 2555018 TI - Small hepatocellular carcinoma: detection with sonography, computed tomography (CT), angiography and Lipiodol-CT. AB - Seventy-three small hepatocellular carcinomas under 5 cm in diameter in 47 patients were examined by sonography, computed tomography (CT), hepatic angiography and CT after intra-arterial injection of iodized poppy-seed oil (Lipiodol-CT). The imaging techniques that first led to detection of small hepatocellular carcinomas were sonography in 53 cases (72.6%), CT in 10 (13.7%), angiography in eight (11%) and Lipiodol-CT in two (2.7%). Sensitivity for detecting small hepatocellular carcinomas was 73% with sonography, 82% with CT, 86% with angiography and 96% with Lipiodol-CT. As a screening method, sonographic and CT results in detecting small hepatocellular carcinomas were not significantly different (p greater than 0.05). Lipiodol-CT was superior to sonography (p less than 0.01), CT (p less than 0.01) and angiography (p less than 0.05) in detecting small hepatocellular carcinomas. We believe that the combined use of Lipiodol-CT with screening methods such as sonography or CT is indispensable for the accurate detection of small hepatocellular carcinomas. PMID- 2555017 TI - Interaction of the Paf antagonist WEB 2086 and its hetrazepine analogues with human platelets and endothelial cells. AB - 1. Intact platelets and confluent human umbilical vein endothelial cells bound [3H]-Paf-acether (platelet activating factor, [3H]-Paf) at 20 degrees C in the presence of 0.25% (w/v) bovine serum albumin (BSA). 2. [3H]-Paf binding to platelets was inhibited in a concentration-dependent manner by WEB 2086. An excess of WEB 2086 indicated the presence of specific, saturable Paf binding which reached a maximum of 28.3 +/- 3.7 fmol [3H]-Paf per 5 x 10(7) platelets. In platelets, different hetrazepines (WEB 2098, 2105, but not 2118) also inhibited [3H]-Paf binding in a concentration-dependent manner. 3. WEB 2086 partially displaced platelet-bound [3H]-Paf in a concentration-dependent manner reaching a plateau at 400 nM WEB 2086. No further displacement was observed when WEB 2086 and an excess of unlabelled Paf were added together. 4. The hetrazepines inhibited platelet aggregation. Platelet aggregation IC50 values correlated well with the IC50 values of the hetrazepines against [3H]-Paf binding (r2 = 0.99). WEB 2086 shifted the Paf dose-response curve rightwards in a parallel manner. Tested against platelet aggregation the pA2 obtained for WEB 2086 was 7.9. 5. WEB 2086 inhibited [3H]-Paf binding to endothelial cells in a concentration-dependent manner. WEB 2086 also inhibited the Paf-mediated cytosolic calcium increase in endothelial cells with an IC50 value of 23.1 +/- 10.4 nM as compared with an IC50 of 21.6 +/- 10.4 nM WEB 2086 for platelet aggregation. 6. These results demonstrate an inhibition of [3H]-Paf binding to platelets and endothelial cells by different hetrazepines, most probably at the Paf receptor level. PMID- 2555019 TI - Primary malignant fibrous histiocytoma of a metacarpal bone: a new localization. PMID- 2555020 TI - AIDS presenting as cytomegalovirus cystitis. PMID- 2555016 TI - Do both adrenaline and noradrenaline stimulate cardiac alpha-adrenoceptors to induce positive inotropy of rat atria? AB - 1. The positive inotropic responses of rat paced left atria to adrenaline and noradrenaline were recorded. Desmethylimipramine (DMI, 1 microM) and metanephrine (10 microM) were initially present throughout. 2. The positive chronotropic responses of spontaneously beating right atria to adrenaline were used as a reference. In these, pindolol, in increasing concentrations, caused progressive shift of the concentration-response curves to the right, which yielded a pA2 value (8.15) compatible with antagonism of beta-adrenoceptors. 3. The left atrial tension responses to adrenaline showed an initial progressive displacement by pindolol (up to 3 microM) which gave an unexpectedly low pA2 value (6.48). However, with further increases in pindolol concentration there was no additional shift of the curve. In the presence of pindolol (3 microM), prazosin (0.1 microM) displaced the curve to the right but the pA2 value derived from this shift (7.75) was less than expected for alpha 1-adrenoceptor antagonism. 4. When the experiments in the presence of pindolol (3 microM) were repeated in the absence of DMI, prazosin displaced the concentration-response curves for adrenaline induced left atrial tension to a greater extent and the pA2 value (8.76) was now compatible with adrenaline stimulating typical alpha 1-adrenoceptors. 5. The concentration-response curves for noradrenaline-induced left atrial tension were also progressively displaced to the right by pindolol (0.1, 0.3 and 1.0 microM). These concentrations yielded a Schild plot of unity slope and a pA2 value of 7.94 +/- 0.04. This was not significantly different from the pA2 value of 8.02 +/- 0.07 determined for pindolol against isoprenaline in the left atria, which indicates a normal interaction of noradrenaline with beta-adrenoceptors in the absence and presence of low concentrations of pindolol. 6. A further increase in the concentration of pindolol to 3 microM failed to induce an additional shift of the noradrenaline curves, whether a 'before and after' antagonist or a 'naive tissue' design was adopted. Similarly, the rightwards shift of the concentration response curves by timolol reached a limit as the concentration was increased. In all cases the limit of shift occurred at a noradrenaline EC50 value of 5-10 microM. 7. At the limit of beta-adrenoceptor antagonism, prazosin and dibenamine did not displace the noradrenaline curves further. The residual inotropic response to noradrenaline therefore appeared to be mediated via neither alpha- nor beta-adrenoceptors. 8. DMI, in the absence of beta-blockade, produced the potentiation of adrenaline and noradrenaline expected of a neuronal uptake inhibitor. However, in the presence of pindolol, there was no potentiation of the right atrial rate response to adrenaline while its left atrial tension responses were antagonized. This suggested that DMI was acting as an alpha-adrenoceptor antagonist. It also explained the less-than-expected shift by prazosin of the adrenaline responses in the presence of both pindolol and DMI, the latter drug already exerting some alpha-blocking activity. In contrast, the left atrial tension responses to noradrenaline in the presence of pindolol (1 microM) were neither potentiated nor antagonized by DMI. 9. When the effects of prazosin upon left atrial tension responses to noradrenaline in the presence of pindolol (10 microM) were examined in the presence of a lower concentration of DMI (O.1 microM) or cocaine (1O microM), again there was no further shift of the curve. However, when the effect of prazosin) The Macmillan Press Ltd 1989 598 K.L. WILLIAMSON & K.J. BROADLEY was examined in the absence of DMI, but in the presence of pindolol (1 and 1O microM) or timolol (3 microM), there was a small shift of the curves by prazosin (0.1 microM). This yielded pA2 values of 7.19, 7.34 +/- 0.1 and 7.66 +/- 0.09, which were at least one order of magnitude less than literature values and that obtained with adrenaline (8.76 +/- 0.18), and are not consistent with noradrenaline stimulating an alpha 1-adrenoreceptor in the presence of beta-adrenoceptor blockade, the increase in left atrial tension by noradrenaline does not appear to be mediated by beta l- or typical alpha adrenoceptors. This is in contrast to adrenaline which in these conditions stimulates typical alpha 1-adrenoceptors. PMID- 2555021 TI - Ontogenesis of diurnal rhythms of cAMP concentration, outer segment disc shedding and retinomotor movements in the eye of the brown trout, Salmo trutta. AB - Histological analyses of the retina of Salmo trutta post-embryos exhibit rhythmic disc shedding at 6 weeks, followed by cyclic retinomotor movements at 7 weeks. Diurnal variations in cAMP concentration, as established by a radioreceptor assay, develop simultaneously with retinomotor movements and are fully expressed at 9.5 weeks. In continuous darkness, neither retinomotor movements nor intra ocular peaks of cAMP occur, indicating that the endogenous oscillator for retinomotor movements and cAMP levels is not yet developed. PMID- 2555022 TI - Characterization of [125I]HEAT binding to alpha 1-receptors in human brain: assessment in aging and Alzheimer's disease. AB - The binding of [125I]2-(beta-4-hydroxyphenylethylamino-ethyltetralone ([ 125I]HEAT), an alpha 1-adrenergic receptor antagonist, to human brain membranes was characterized and the binding assessed in tissue from subjects with Alzheimer's disease (AD) and aging controls. Under Na+-K+ phosphate buffer conditions, [125I]HEAT bound to a single class of binding sites in prefrontal cortex (Brodmann area 10) with a Kd of about 120 pM. High binding capacities of [125I]HEAT were evident in the hippocampus and neocortex but were low in subcortical areas and cerebral microvessels comparable to the regional distribution of [3H]prazosin binding reported previously. Displacement of [125I]HEAT by various adrenergic drugs was consistent with its binding to alpha 1 adrenergic receptors. The specific binding was not affected by postmortem delay between death and freezing of tissue at autopsy. There was no correlation of [125I]HEAT binding with age of subjects. In AD subjects, the binding was significantly decreased in prefrontal cortex by about 25% but not changed in hippocampus, putamen or cerebellum compared to age-matched controls. The reduced binding of [125I]HEAT in prefrontal cortex may reflect a region-specific change in alpha 1-adrenergic receptors associated with neuronal loss in AD. PMID- 2555023 TI - Chronic exposure of developing cortical neurons to GABA down-regulates GABA/benzodiazepine receptors and GABA-gated chloride currents. AB - Cultures of cerebral neurons were prepared from chick embryos, 8.5 days in ovo, and maintained in vitro. Following chronic exposure of these cells to GABA, the levels of [3H]flunitrazepam binding in situ and electrophysiological responsiveness to gamma-aminobutyric acid (GABA) was examined. Treatment with 100 microM GABA for 7 days reduced [3H]flunitrazepam binding in situ by 70 +/- 8% compared to untreated controls. The binding of [3H]N-methylscopolamine was unaffected by this treatment. The reduction in [3H]flunitrazepam binding was prevented by concomitant exposure of developing neurons to the GABA antagonist R 5135, suggesting that GABAA receptor occupancy is required. The loss of bezodiazepine receptors was dependent on the GABA concentration in the culture medium and a half-saturation (IC50) value of 11.2 +/- 3.7 microM was estimated. Whole-cell patch-clamp recordings were obtained to assess the functional properties of the labile receptor pool observed in the binding studies. Neurons cultured with 100 microM GABA for 7 days showed a 60-70% reduction in the peak current amplitudes observed in response to application of 10-100 microM GABA. However, the rate of rapid desensitization, quantified by measuring changes in input conductance, was unchanged by chronic GABA exposure, yielding decay time constants of 27.1 +/- 2.1 and 34.7 +/- 4.7 s for control and treated cells, respectively. The results are consistent with a GABA modulation of the GABAA/benzodiazepine receptor complex by means of down-regulation. PMID- 2555024 TI - Stimulation of kappa opiate receptors in intestinal wall affects stress-induced increase of plasma cortisol in dogs. AB - In dogs, an acoustic stress (A.S.) produced by hearing of intense music (less than or equal to 90 dB) through earpieces for 1 h induced a 520% maximal rise in plasma cortisol 15-30 min after the beginning of stress. Oral administration of the specific kappa agonists, U-50488 (0.1 mg/kg) and PD 117302 (0.05 mg/kg), 30 min before the A.S. session reduced significantly (P less than 0.01) by 71.2% and 80.9% the maximal increase of plasma cortisol but did not affect the increase observed after intracerebroventricular administration of ovineCRF (100 ng/kg). These effects which are not reproduced by intravenous administration of the drugs at similar doses, were blocked by previous treatment with MR 2266 (0.1 mg/kg) or local anesthesia and vagotomy, suggesting that kappa opioid agonists inhibit the stress-induced activation of the hypothalamo-pituitary-adrenocortical (HPA) system by acting selectively on specific receptors located in the wall of the proximal gut. PMID- 2555025 TI - Correlation of ontogeny with function of [3H]U69593 labelled kappa opioid binding sites in the rat spinal cord. AB - In this study, we have used a variety of in vitro and in vivo techniques to demonstrate the presence, and examine the function, of [3H]U69593 binding sites in the spinal cord of the 9-16-day-old rat in comparison to the adult. Equilibrium binding of [3H]U69593 to homogenates of adult rat spinal cord revealed a single population of non-interacting sites with a maximum binding capacity of 10.4 +/- 1.4 fmol/mg protein and an apparent equilibrium dissociation constant of 2.31 +/- 0.47 nM while in 9-16-day-old cord these parameters were 57.0 +/- 9.4 fmol/mg protein and 2.28 +/- 0.22 nM, respectively. The total binding capacity per cord was 95.8 +/- 8.3 and 121.8 +/- 7.7 fmol/cord for adult and immature rat, respectively. Competition studies using receptor-selective opioid ligands showed that these sites were kappa opioid in nature. Autoradiographical techniques demonstrated a uniform distribution of these sites over transverse sections of 9-16-day-old rat cord. In vitro electrophysiology was performed on spinal cord slice preparations from the 9-16-day-old rat. U69593 (100 nM-1 microM) had no effect on passive membrane properties but produced a naloxone-reversible depression of both spontaneous and electrically evoked activity in dorsal horn neurones. Direct intrathecal injection of U69593 (0.3 10.0 micrograms/animal) into 9-16-day-old rats produced a dose-dependent, naloxone-reversible, antinociception when measured using the paw-pressure test. In conclusion, we have shown that, in contrast to the adult, the spinal cord of the 9-16-day-old rat has a significantly higher concentration of [3H]U69593 binding sites which have functional in vitro and in vivo correlates. PMID- 2555026 TI - Pharmacological sensitivities of two afterhyperpolarizations in rat optic nerve. AB - Optic nerves were removed from 4-8-week-old rats and studied in a modified sucrose gap chamber in order to examine the pharmacological sensitivities of two afterhyperpolarizations (AHPs), an early and a late one. The peak latency of the early AHP, which occurred immediately after the action potential, was 6-12 ms, and its duration was 44-75 ms. The early AHP was present at resting potential in about 75% of recorded nerves. In the nerves in which an early AHP was not present at resting potential, an AHP was present during delivery of constant current depolarization through the sucrose gap. The early AHP was increased in amplitude by depolarization of the nerve, decreased in amplitude or reversed in polarity by hyperpolarization, was not affected by tetraethylammonium (TEA) or Co2+, and was obliterated by 4-aminopyridine (4-AP) concomitant with action potential broadening. The late AHP followed a broadened action potential in the presence of the potassium channel blocker 4-AP. Its peak latency ranged from 112 to 254 ms and its duration from 336 to 710 ms. It was increased in amplitude with repetitive stimulation, was reversibly obliterated by TEA, but was not significantly altered by Co2+, Cd2+, Ca2+ removal, charybdotoxin or apamin. The results demonstrate the presence of two AHPs mediated by pharmacologically distinct potassium conductances in rat optic nerve, neither of which is calcium dependent. PMID- 2555027 TI - Disinhibitory actions of the GABAA agonist muscimol in immature hippocampus. AB - Bath application of muscimol (10-20 microM) to hippocampal slices obtained from rats on postnatal days 10-15 produced epileptiform activity in the form of multiple population spikes in 20% of slices tested, concurrent with marked disinhibition. The disinhibition occurred in nearly 100% of cases tested at muscimol concentrations that produced epileptiform activity. Paired pulse analysis of GABAergic recurrent inhibition revealed a muscimol-induced disinhibitory effect involving a decrease in maximum possible inhibition. Spontaneous and antidromically elicited inhibitory postsynaptic potentials (IPSPs) recorded intracellularly were suppressed by muscimol. Current-voltage analysis of the recurrent IPSPs suggests that muscimol acted at a number of sites to produce disinhibition. The input conductance of the postsynaptic pyramidal cell increased due to muscimol, creating a current shunt which likely decreased the efficacy of synaptic currents. Muscimol also caused a decrease in the conductance due to the IPSP as well as a shift in the depolarizing direction of the equilibrium potential of the IPSP. The data indicate that muscimol, a GABAA agonist, can produce disinhibition resulting from the multiple consequences of its action. We conclude that the physiologic consequences of GABAA agonist treatment are complex. On the other hand, neurons are likely to be inhibited by a tonic increase in membrane conductance. However, since recurrent inhibition is simultaneously compromised, excitatory vollies of sufficient intensity may overcome the tonic inhibition and produce a hyperexcitable state. In some cases, this disinhibition may induce epileptiform activity. These observations are relevant in light of the proposed use of GABA agonists clinically to control seizures. PMID- 2555028 TI - A quantitative description of excitatory amino acid neurotransmitter responses on cultured embryonic Xenopus spinal neurons. AB - We have performed a quantitative analysis of excitatory amino acid neurotransmitter receptors on cultured embryonic Xenopus spinal neurons using the whole-cell patch-clamp technique. Neuroblasts and underlying mesodermal cells isolated from spinal regions of neural plate-stage embryos were placed into dissociated cell culture, and responses were studied soon after the appearance of neurites on embryonic neurons. Glutamate (Glu) receptors were separated into two general classes based on responses to the characteristic agonists quisqualate (Quis), kainate (Ka) and N-methyl-D-aspartate (NMDA); these were NMDA receptors (those activated by NMDA) and non-NMDA receptors (those activated by Ka and Quis). Half-maximal responses to Glu and other agonists on NMDA and non-NMDA receptors were determined from Hill analysis of dose response relations. The order of sensitivities observed was: GluNMDA (ED50 = 5.1 microM) greater than Glunon-NMDA (ED50 = 28 microM), and for Glu receptor agonists, Quis (ED50 = 1.5 microM) greater than NMDA (ED50 = 41 microM) greater than Ka (ED50 = 58 microM). The order of response amplitudes recorded at concentrations near the appropriate ED50s was GluNMDA greater than Glunon-NMDA, and Ka greater than NMDA greater than Quis. A 10-fold decrease in external [Na+] shifted the reversal potentials for Glunon-NMDA, Ka, and Quis to more negative voltages. Increasing external [Ca2+] shifted the reversal potential for NMDA responses to more positive potentials, an observation consistent with Ca2+ permeation of the embryonic NMDA-activated channel. NMDA-evoked currents could not be recorded in nominally glycine (Gly) free media. Addition of Gly to external solutions potentiated NMDA responses (ED50 = 644 nM). NMDA responses were blocked by DL-2-amino-5-phosphonovaleric acid (APV; ED50 = 1.9 microM) and by Mg2+ at negative potentials. In their sensitivities to agonists and antagonists, and ionic dependences, amino acid neurotransmitter responses on embryonic Xenopus neurons closely resembled those previously observed for mature Xenopus and mammalian central neurons. The GluNMDA receptors present on these immature neurons were sufficiently sensitive to be activated by endogenous concentrations of extracellular Glu, suggesting a possible role for receptor activation in modulating early neural development. PMID- 2555029 TI - Acetylation alters the feeding response to MSH and beta-endorphin. AB - The effects on food intake of the N-acetylation of MSH and beta-endorphin have been examined following their injection into the third ventricle. Desacetyl-MSH and alpha-MSH were injected into fasted rats, and beta-endorphin and N-acetyl beta-endorphin into fed rats. Desacetyl-MSH had no effect on food intake following ICV injection into food-deprived rats at any dose between 100 and 2500 pmoles. Alpha-MSH, the N-acetylated form of MSH, on the other hand, showed a highly significant inhibition of food intake in food-deprived rats with doses of 100 and 250 pmoles but no effect with the higher doses. With beta-endorphin, there was a dose-related biphasic effect. One hour after injection of beta endorphin (2500 pmole) food intake was inhibited whereas the lowest dose, 100 pmole, significantly stimulated it. By 3 hours, the 2 lowest doses of beta endorphin both significantly stimulated food intake, but the highest dose remained inhibitory. By 6 hours all doses of beta-endorphin stimulated food intake compared to the vehicle-treated animals. In contrast, N-acetylation of beta-endorphin eliminated all effects on food intake following injection into the third ventricle. These data suggest that N-acetylation of products formed by the processing of POMC can markedly alter their biological properties. PMID- 2555030 TI - Persistent alterations in GABAA receptor binding and function after prenatal lorazepam administration in the chick. AB - Behavioral abnormalities have been reported in young and adult animals exposed to benzodiazepines prenatally. The presence of neurochemical alterations in the GABAergic system after prenatal benzodiazepine exposure was assessed in a chick model which avoids prenatal and postnatal maternal effects. The GABAA receptor complex, the presumed site of benzodiazepine action, was altered in adult chickens previously exposed to lorazepam for 10 days in ovo. Binding was decreased at the putative chloride channel site labeled by [35S]TBPS, coupling was decreased between this site and the GABA binding site, and function of the GABAA receptor in chloride uptake was diminished in animals exposed to prenatal lorazepam. Persistent neurochemical alterations in the GABAergic system accompany prenatal benzodiazepine exposure, and may influence subsequent behavior and development. PMID- 2555031 TI - Estradiol or ovariectomy decreases CRF synthesis in hypothalamus. AB - Gonadectomy and gonadectomy plus chronic estradiol administration decrease the content of hypothalamic corticotropin-releasing factor-like immunoreactivity (CRF ir). This investigation was conducted to determine whether this was a result of chronic inhibition of CRF synthesis or stimulation of release. Administration of the protein synthesis inhibitor anisomycin one hour prior to sacrifice in otherwise untreated controls resulted in a 21% decrease in median eminence CRF-ir (p less than 0.01). Rats were ovariectomized and then administered either estradiol or vehicle daily for 3 weeks. Compared to vehicle-treated controls, estradiol treatment decreased CRF-ir in median eminence (p less than 0.01). In contrast to controls, administration of anisomycin to these ovariectomized rats did not significantly decrease median eminence CRF-ir in either the vehicle or estradiol-treated groups, implying that CRF synthesis was already depressed. Plasma adrenocorticotropin (ACTH) was increased by anisomycin treatment in all groups whether intact, ovariectomized or estradiol-treated (p less than 0.0005). Adrenal weights did not differ between these groups, or in comparison to sham ovariectomized rats, indirectly implying lack of chronically elevated plasma ACTH. These data suggest that the mechanism for both the ovariectomy and chronic estradiol-induced decrease in CRF-ir content is an inhibition of CRF synthesis as opposed to a stimulation of CRF release. PMID- 2555032 TI - Developmental changes in the corticosterone response to corticotrophin and in the adrenal corticosterone content of rapid and slow growing strains of chickens (Gallus domesticus). AB - 1. The high sensitivity of the immature domestic fowl to corticotrophin (ACTH) has been confirmed. The minimal effective dose was age-dependent, and decreased from 2 IU/kg BW in 1-d-old chicks to 0.4 IU/kg BW in 12-d-old chicks. 2. Increasing doses of ACTH evoked similar maximal corticosterone responses but the time to return to basal values increased with increasing dose of ACTH in both a broiler and layer strain. 3. Maximal corticosterone responses to ACTH occurred earlier in hatched chicks compared with embryos, and were delayed in embryos and young chicks of the layer strain compared to those of the broiler strain. 4. Strain differences in basal concentrations of corticosterone as well as in the response to ACTH occurred at a time when there were differences in the corticosterone content of the adrenals. PMID- 2555033 TI - [Partial deficiency in 21-hydroxylase in certain forms of hirsutism]. AB - The ACTH test is important when hirsutism occurs in women with a slight 21 hydroxylase deficiency, and normal basal 17-OH Progesterone (17-OH-P/plasma levels). Extensive hormonal assays: LH, FSH, Prolactin, 17 beta-estradiol (E2), Estrone, 17OH-P, Androstenedione, Testosterone, Cortisol (C), Dehydroepiandrosterone-S (DEA-S) were carried out in 36 hirsute women. 13 of these presented hormone levels as found in polycystic ovary syndrome (PCOS), 6 women presented a slight 21-hydroxylase deficiency (increased plasma 17-OH-P and decreased C after ACTH test with significant, p less than 0.01, increase of 17-OH P/C and 17 women presented idiopathic hirsutism (IH). The hormonal pattern, in the basal condition, is not different in IH or in slight 21-hydroxylase deficiency. The ACTH test is able to differentiate between IH and adrenal hirsutism. PMID- 2555034 TI - [Activation in vivo of the collagenase of Clostridium histolyticum by erythromycin lactobionate]. AB - The authors demonstrate a previously not described increase, in vivo, in a mean proportion of 70%, of the enzymatic activity of the clostridium collagenase in rats by erythromycin lactobionate. The pathogenesis of this phenomenon cannot yet be explained. PMID- 2555035 TI - Fatal angioedema associated with enalapril. AB - A 65-year-old male with a long history of congestive heart failure presented to the emergency room with facial swelling 7 h after the first dose of enalapril. He was treated with diphenhydramine and hydrocortisone which improved his angioedema. However, there was a profound relapse of the angioedema followed by respiratory arrest. He suffered severe anoxic brain damage and died five days later. It is likely that the long half-life of enalapril lead to this rebound phenomenon. Enalapril induced angioedema is reviewed with suggestions for management. PMID- 2555036 TI - Concentration of Giardia lamblia cysts, Legionella pneumophila, Clostridium perfringens, human enteric viruses, and coliphages from large volumes of drinking water, using a single filtration. AB - Poliovirus, coliphages, Giardia lamblia cysts, Clostridium perfringens spores, and Legionella pneumophila were concentrated simultaneously in a single pass by sequential filtration of large volumes of drinking water through 3- and 1-micron wound electronegative fiberglass cartridge filters (25.4 cm). Filtration was performed under acidic conditions (pH 3.5) in the presence of 0.001 M aluminum chloride to enhance adsorption. Elution of all the microorganisms entrapped or adsorbed to the filters was obtained by a slow backwash elution with a 1.5% beef extract solution, pH 9.75, containing 0.5% Tween 80. Tween 80 was shown to enhance recovery of the bacteriophages, bacteria, and parasites. Giardia cysts were efficiently eluted (71%) and could be reconcentrated by low-speed centrifugation and purified by sucrose density gradient flotation at a final recovery of 52%. Legionella pneumophila cells were eluted at 64% and were further concentrated by low-speed centrifugation at an overall recovery of 55%. C. perfringens spores and coliphages were eluted at efficiencies of 82 and 86%, respectively, and reconcentrated with minimal loss by a detergent - protein flotation method. Poliovirus was eluted at 93% and reconcentrated at 78% efficiency by organic flocculation. PMID- 2555037 TI - Genetic mapping of katB, a locus that affects catalase 2 levels in Bacillus subtilis. AB - A locus affecting the synthesis of spore-specific catalase 2 in Bacillus subtilis was mapped using two- and three-factor transductional crosses at 342 degrees between hsrE and iol. It was named katB. Strains lacking catalase 2 remained sporulation proficient, but blockage of sporulation at stage IV or earlier affected the electrophoretic mobility of the native enzyme. PMID- 2555038 TI - Reevaluation of procarbazine for the treatment of recurrent malignant central nervous system tumors. AB - Ninety-nine patients with primary recurrent malignant tumors of the central nervous system were treated with procarbazine as a single agent. Procarbazine was not given as a specified protocol, but for patients who were ineligible or refused other protocols. All patients had been treated previously with radiotherapy and 96 patients had also received previous chemotherapy. Twenty-five patients were treated at the first progression of their tumor, 47 were treated at the second progression, and 27 were treated at the third progression of their tumor. For the aggregate, the response plus stabilization rate was 27% for glioblastoma multiforme with median time to tumor progression of 30 weeks, and 28% for other anaplastic gliomas with a median time to tumor progression of 49 weeks. With respect to the percent of patients who responded or stabilized to treatment, these results are inferior to those reported previously for patients treated with procarbazine at recurrence. With respect to duration of response and stabilization, the data are comparable. PMID- 2555039 TI - Intraarterial infusion of 5-fluoro-2-deoxyuridine-C8 dissolved in a lymphographic agent in malignant liver tumors. A preliminary report. AB - The drug 5-fluoro-2-deoxyuridine-C8 (FUdR-C8), one of the lipophilic prodrugs of FUdR, was dissolved in an oily lymphographic agent (Lipiodol Ultra Fluid, Andre Gelbe Laboratory, Paris, France; Ethiodol, Savage Laboratories, Melville, NY) and used for the intraarterial treatment of malignant liver tumors. From August 1985 to June 1988, 33 patients with hepatocellular carcinoma and 13 patients with metastatic liver tumors were treated with this agent at the Kumamoto University Hospital and its affiliated hospitals. The response rate (complete remission [CR] and partial remission [PR]) was 27.6% for hepatocellular carcinomas and 46.1% for metastatic liver tumors. The cumulative 1-year survival rate was 55.1% for hepatocellular carcinomas and 70.0% for metastatic liver tumors. More than a 50% decrease in the tumor marker level was observed in ten of 21 patients with hepatocellular carcinoma and in two of eight patients with metastatic liver tumors. The side effects, which were transient and controlled with conservative treatment, included fever, abdominal pain, nausea, vomiting, and acute gastritis. PMID- 2555040 TI - Evaluation of des-gamma-carboxy prothrombin as a marker protein of hepatocellular carcinoma. AB - We measured des-gamma-carboxyglutamic acid prothrombin (protein induced by vitamin K absence or antagonist-Factor II: [PIVKA-II]) in plasmas of normal subjects, patients with thrombotic disease, those with hepatic disease including hepatocellular carcinoma, and those with carcinoma of other tissues, and compared the results with results of blood coagulation tests used for the examination of hepatic function. In addition, in the patients with hepatic disease, PIVKA-II and alpha-fetoprotein (AFP) levels were compared. The PIVKA-II level was frequently high in patients with thrombotic disease given warfarin therapy and those with hepatocellular carcinoma. However, in patients with thrombotic disease who were not given warfarin therapy, no significant correlation was seen between the PIVKA II value and the results of the thrombotest or hepaplastin test, suggesting no association between the PIVKA-II level and the degree of impairment of hepatic function. In 70 patients with hepatocellular carcinoma, the percentage of patients positive for PIVKA-II (greater than or equal to 0.1 micrograms/ml) and those positive for AFP (greater than or equal to 20 ng/ml) were similar (77% and 74%, respectively). Pearson's correlation of coefficient between the PIVKA-II value and the AFP value in the 70 patients was 0.463. However, false-positive rates in patients with hepatic disease other than hepatocellular carcinoma were lower for PIVKA-II. Combined assessment of PIVKA-II and AFP increased positive rates and allowed exclusion of false-positive patients. The plasma PIVKA-II level is suggested to be useful as an indicator of warfarin control in patients with thrombotic disease, as a marker of hepatocellular carcinoma, and is particularly of value when assessed in combination with AFP. PMID- 2555041 TI - Neurologic complications in children with soft tissue and osseous sarcoma. AB - From 1980 to 1987, 162 consecutive children with soft tissue and osseous sarcoma were reviewed to determine the frequency and types of neurologic complications seen. Neurologic complications occurred in 43 of 162 (26.5%) patients. Children with poorly differentiated sarcomas and rhabdomyosarcoma were more likely to have neurologic complications, which occurred in 39% of patients at risk. The types of complications seen included: metastatic spinal cord compression (11%); symptomatic peripheral neuropathy (10%); intracranial metastatic disease (7.5%); seizures (6%); and acute and chronic methotrexate-related neurologic dysfunction (2.5%). Spinal cord compression frequently occurred early in disease whereas brain metastases was almost always a late finding. Symptomatic peripheral neuropathy occurred primarily in children with rhabdomyosarcoma and Ewing's sarcoma. The advent of increasingly successful therapies for children with sarcoma and the frequency of severe neurologic complications indicate that a heightened level of surveillance for neurologic compromise is required. PMID- 2555042 TI - Tumor promoters stimulate the formation of 1,2-diacylglycerol in murine peritoneal macrophages: a possible mechanism for stimulating superoxide anion radical production. AB - The formation of 1,2-diacylglycerol (DAG), a known stimulator of superoxide anion radical (O2-.) production in inflammatory cells, was assessed in murine peritoneal macrophages following treatment in vitro with tumor promoters. Addition of phorbol-12-myristate-13-acetate (PMA, 1-100 ng/ml) to resident peritoneal macrophage cultures from CD-1 female mice resulted in a 3- to 7-fold increase in [3H]DAG formation. The response was observed from 15 min to 2 h following the addition of PMA. At concentrations at which they stimulate O2-. production, PMA and other tumor promoters such as mezerein, phorbol-12,13 dibutyrate and 4-O-methyl-PMA stimulated the formation of [3H]DAG. Similar results were obtained when thioglycollate-elicited macrophages were used. Concurrent with the formation of [3H]DAG was a release of [3H]choline equivalents from the resident peritoneal macrophages treated with tumor promoters. The calcium ionophore A23187 did not stimulate O2-. production of [3H]DAG formation in resident peritoneal macrophages. These results demonstrate that tumor promoters stimulate the accumulation of DAGs in murine peritoneal macrophages at concentrations at which they stimulate O2-. production and suggest a mechanism by which tumor promoters such as mezerein, which are weak activators of protein kinase C, may indirectly stimulate O2-. production. PMID- 2555044 TI - A response in advanced post-menopausal breast cancer during treatment with the luteinising hormone releasing hormone agonist--Zoladex. AB - Luteinising hormone releasing hormone (LHRH) agonists are currently undergoing clinical trials in the treatment of advanced breast cancer in pre-menopausal women. Clinical responses are attributed to the suppression of the pituitary ovarian axis, with a reduction in circulating levels of gonadal steroids similar to that produced by castration. In the present case report, we report a partial response to a LHRH analogue in a post-menopausal woman refractory to other endocrine treatments. This response cannot be explained with a chemical castration and confirms the possible direct anti-tumor effect of Zoladex. PMID- 2555043 TI - Modulation of galactoside-binding lectins in tumor cells by differentiation inducing agents. AB - The effects of the differentiation-inducing agents N6, O2'-dibutyryl cyclic AMP, beta-all-trans retinoic acid, dimethylsulfoxide and butyrate on the levels of galactoside-binding proteins (lectins) in cultured human and murine tumor cells were examined by immunoblotting. Differentiation was associated with decreased levels of a 34-kDa lectin in the K-1735P and B16-F1 melanoma cells and decreased levels of a 14.5-kDa lectin in S20 neuroblastoma, MDA-MB 175 breast carcinoma, HL 60 and THP-1 leukemia cells. The level of a 14.5-kDa lectin increased during differentiation of F-9 embryonal and KM12P colon carcinoma cells. These results indicate that tumor cell differentiation along specific pathways is accompanied by distinct modulation of lectin expression. These changes may recapitulate the normal developmental regulation of lectin expression. PMID- 2555045 TI - Isolation and characterization of a hepatoma-associated abnormal (des-gamma carboxy)prothrombin. AB - Hepatoma-associated abnormal (des-gamma-carboxy)prothrombin (HAPT) is a newly described tumor marker for hepatocellular carcinoma. HAPT has been measured in the blood of patients with hepatoma by immunoassay but has not been isolated or characterized. This paper describes the quantitative isolation and structural characterization of HAPT. Purified HAPT has the same molecular weight, amino terminal sequence, and amino acid analysis (exclusive of gamma-carboxyglutamic acid) as native prothrombin and abnormal prothrombin isolated from the blood of patients taking sodium warfarin. HAPT is heterogeneous in gamma-carboxyglutamic acid (Gla) content with an average of 5 Gla residues/molecule compared to 10 Gla residues for native prothrombin and 2 Gla residues for abnormal prothrombin. HAPT is glycosylated in a manner equivalent to that for native prothrombin when evaluated by a concanavalin A-binding assay. These studies find structural identity between HAPT and abnormal prothrombin. Therefore the findings support the hypothesis that HAPT results from an acquired defect in the posttranslational vitamin K-dependent carboxylation of the prothrombin precursor and not an intrinsic defect in the prothrombin precursor molecule. PMID- 2555047 TI - Phosphorylation of 3-deazaguanosine by nicotinamide riboside kinase in Chinese hamster ovary cells. AB - The growth inhibitory activity of 3-deazaguanosine toward a mutant line (TGR-3) of Chinese hamster ovary cells deficient in hypoxanthine-guanine phosphoribosyltransferase (EC 2.4.2.8) was substantially reversed by the simultaneous addition of nicotinamide riboside. The activities of most other ribonucleoside analogues tested were unaffected. The formation of cellular 3 deazaGMP and 3-deazaGTP from the ribonucleoside analogue, as measured by high pressure liquid chromatography, was inhibited by the presence of nicotinamide riboside. The inhibition was dependent on concentration of 3-deazaguanosine and could also be demonstrated by following the metabolism of 3-deazaguanosine, labeled with 14C in the ribose moiety, to [14C]3-deazaGTP. In the presence of 100 microM nicotinamide riboside formation of the labeled triphosphate derivative of 3-deazaguanosine was undetectable. A 3-deazaguanosine phosphorylating activity was separated from other cellular kinases by DEAE-cellulose chromatography. Contaminating purine nucleoside phosphorylase (EC 2.4.2.1) was subsequently removed by sucrose density gradient centrifugation. The resulting enzyme preparation demonstrated the greatest activities with nicotinamide riboside and 3 deazaguanosine and, in addition, could also phosphorylate tiazofurin and guanosine to lesser, but significant, degrees. These and other observations suggest that 3-deazaguanosine, and perhaps other agents such as tiazofurin, may, at least in part, be phosphorylated by a nicotinamide ribonucleoside kinase in these cells. If so, it is possible that the activity of this agent in other types of cells in vivo could be dependent upon the presence of this enzyme and that it could be influenced by cellular concentrations of the natural pyridine nucleoside. PMID- 2555046 TI - Heat- or stress-inducible transformation-associated cell surface antigen on the activated H-ras oncogene-transfected rat fibroblast. AB - A WKA rat fetus fibroblast (WFB) was transfected by several oncogenes including EJras (activated H-ras), polyoma middle T (PyMT), v-src, c-myc, and adenovirus type 12 E1A-E1B. We analyzed the expression of the transformation-associated cell surface antigens on WFB by developing monoclonal antibodies. One of the WFB transformation-associated cell surface antigens, recognized by monoclonal antibody 067, was constitutively expressed only on two (W31 and W70) of ten WFB EJras-transformed clones. This antigen could not be detected on parental WFB cells as well as ten WFB-PyMT transformant clones. Furthermore, it was not expressed on several clones of partially transformed WFB-v-src and WFB-adeno E1 transfectants or nontransformed WFB-c-myc transfectants. Monoclonal antibody 067 could form an immunoprecipitate with an approximate molecular weight of 67,000 which was composed of a single polypeptide chain. It was also shown that the expression of this antigen could be enhanced by cyclic AMP or cholera toxin treatment of W31; treated cells also showed a phenotypic reversion to the nonmalignant growth characteristics of the parental WFB. Moreover, the expression of this antigen could be induced on the WFB-EJras transformants such as W14, which do not constitutively express this antigen, by treatment of these agents. Furthermore, the expression of antigen was enhanced by heat and superoxide treatment on W31. These data suggest that the monoclonal antibody 067-defined molecule is a novel transformation-associated cell surface antigen that could be induced by heat shock or other physiological stress. PMID- 2555048 TI - Diethyldithiocarbamate modulation of murine bone marrow toxicity induced by cis diammine(cyclobutanedicarboxylato)platinum (II). AB - Diethyldithiocarbamate (DDTC) has been shown to ameliorate the myelosuppression induced by the platinum cancer chemotherapeutic drugs in mice. Optimal drug scheduling for DDTC and cis-diammine(cyclobutanedicarboxylato)platinum(II) (CBDCA) has been determined in C57BL/6 x DBA/2 F1 mice, using the pluripotent stem cell assay to assess hematological toxicity. DDTC, at doses of 0.3 to 300 mg/kg given 3 h after 60 mg/kg CBDCA, tripled the number of proliferating spleen colony-forming units compared to treatment with CBDCA alone. No significant difference in efficacy was noted among these doses. DDTC, at the lowest myeloprotective dose (0.3 mg/kg), was most active when administered from 1 to 3 h after CBDCA. The combination of DDTC with CBDCA in vivo did not alter the clonogenic survival of L1210 cells compared to CBDCA alone. CBDCA depressed both bone marrow and tumor cell DNA synthesis. DDTC given 3 h after CBDCA hastened the recovery of DNA synthesis only in marrow cells; the addition of DDTC to CBDCA did not alter DNA synthesis in tumor cells. DDTC alone did not significantly affect DNA synthesis in either normal or tumor cells. These results suggest that the mechanism of DDTC myeloprotection involves stimulation of bone marrow cell proliferation and that the selectivity of DDTC is based upon the absence of stimulation in tumor cells. PMID- 2555049 TI - Gene activation by induced DNA rearrangements. AB - A murine cell line (EN/NIH) containing the retroviral vector ZIPNeoSV(x)1 that was modified by deletion of the enhancer elements in the viral long terminal repeats has been used as an assay system to detect induced DNA rearrangements that result in activation of a transcriptionally silent reporter gene (neomycin phosphotransferase, neo) encoded by the viral genome. The spontaneous frequency of G418 resistance is less than 10(-7), whereas exposure to the tumor promoter 12 O-tetradecanoylphorbol-13-acetate (TPA) or the combination of UV irradiation plus TPA resulted in the emergence of drug resistant cell lines at a frequency of 5 per 10(6) and 67 per 10(6) cells, respectively. In several of the cell lines that were analyzed a low level of amplification of one of the two parental retroviral integrants was observed, whereas in others no alteration in the region of the viral genome was detected. To determine the effect of the SV40 large T antigen on induced DNA rearrangements, EN/NIH cells were transfected with a temperature sensitive (ts) mutant of SV40 T. Transfectants were maintained at the permissive temperature (33 degrees C) for varying periods of time (1-5 days) in order to vary SV40 T antigen exposure, after which they were shifted to 39.5 degrees C for selection in G418. The frequency of emergence of drug resistant cell clones increased with duration of exposure to large T antigen (9-52 per 10(6) cells over 1-5 days, respectively), and all cell lines analyzed demonstrated DNA rearrangements in the region of the neo gene. A novel 18-kilobase pair XbaI fragment was cloned from one cell line which revealed the presence of a 2.0 kilobase pair EcoRI segment containing an inverted duplication which hybridized to neo sequences. It is likely that the observed rearrangement was initiated by the specific binding of large T antigen to the SV40 origin of replication encoded within the viral genome. The investigations with phorbol esters, UV light, and the SV40 large T antigen demonstrate the utility of the EN NIH cell lines for the study of induced DNA rearrangements and support the future use of this system to investigate the mechanism by which varied stimuli or specific gene functions promote DNA rearrangements. PMID- 2555050 TI - Potentiation of nitrogen mustard cytotoxicity by disulfiram, diethyldithiocarbamic acid, and diethylamine in mice. AB - Disulfiram (DSF) and its metabolites, diethyldithiocarbamic acid (DDC) and diethylamine (DEA), were studied as pretreatments in combination with the alkylating agent nitrogen mustard (HN2) on the cytotoxicity of HN2 against both leukemia cells and normal hematopoietic stem cells. Both time intervals and dose relationships were examined. DSF showed a substantial potentiation of HN2 cytotoxicity against murine AKR leukemia cell spleen colony-forming units (LCFU) when given i.p. between 15 to 30 min prior to HN2 i.v. treatment. For 3 mg DSF/mouse pretreatment, leukemia LCFU survival was about 10(-6) whereas it was about 10(-2) for HN2 alone. The extent of this potentiation decreased as the time between treatments increased. Significant potentiation was noted even when a low dose of DSF (0.25 mg/mouse) was administered 15 min before HN2. However, DSF had little if any effect on the modulation of HN2 cytotoxicity to normal hematopoietic cell spleen colony-forming units (NCFU). DDC showed an increasing potentiation of HN2 cytotoxicity against LCFU when given i.p. prior to HN2 i.v. treatment. The maximum effect was noted between 2 and 4 h with a surviving fraction for LCFU between 10(-5) and 10(-6) for 20 mg/mouse DDC pretreatment. The extent of this effect then decreased as the time interval increased beyond 4 h, but it was still significant for the 24-h interval. This pronounced potentiation effect was dose dependent for DDC. The compound exhibited a protective effect against HN2 cytotoxicity to NCFU when given 15 min before HN2. This protection decreased with increased time interval. DEA (20 mg/mouse) did not show a significant potentiation of HN2 cytotoxicity against LCFU when administered i.p. prior to HN2. Also, DEA did not show any significant protection of NCFU. PMID- 2555051 TI - Differentiation of human variant small cell lung cancer cell lines to a classic morphology by retinoic acid. AB - Variant small cell lung cancer (SCLC) is distinguished from the classic histology by changes in growth characteristics and morphology, c-myc amplification, a loss of some biochemical markers, and relative chemo- and radioresistance. Three variant SCLC lines were incubated in 1 microM all-trans retinoic acid. After 8-10 days, a marked change in morphology was noted in all three lines, with tight cell aggregation and central necrosis of large floating spheroids similar to classic SCLC. The retinoid-treated cells demonstrated decreases in growth rate and cloning efficiency to levels comparable with classic SCLC cell lines. Retinoic acid incubation caused a reproducible decrease in c-myc expression in variant SCLC cells, but was not noted to increase L-dopa decarboxylase, an enzyme which biochemically distinguishes classic from variant SCLC cell lines. Retinoid exposure led to an increase in HLA and Leu-7 antigens, but a reduction of antibody binding to 3-fucosyllactosamine, a dominant SCLC glycolipid antigen. Clonogenic assays revealed that the variant cells, after incubation in retinoic acid, became more sensitive to etoposide, but more resistant to Adriamycin. We conclude that exposure of variant SCLC cells to retinoic acid can lead to a morphologic phenotype similar to classic SCLC, but with substantial differences in cell biology. ? PMID- 2555053 TI - Biochemical properties of media conditioned by simian virus 40-induced hamster tumor cells: correlation with distinct cell phenotypes but not with oncogenicity. AB - Hamster cells, transformed in vitro by SV40, have been reported to secrete an unidentified factor(s) that inhibits thymidine uptake (TU) by various normal cell types, including activated lymphocytes. It has been postulated that this apparent antiproliferative effect may play an in vivo role in the high tumorigenic capacity of SV40-transformed hamster cells. In keeping with this hypothesis, Adenovirus type 2-transformed hamster cells, which are only weakly tumorigenic, do not inhibit TU by indicator cells in vitro. To study the biological relevance of this phenomenon, we assayed 11 cell lines derived from different fibrosarcomas, induced in Syrian hamsters by SV40, for their ability to inhibit TU by normal rabbit kidney indicator cells. In contrast to cells transformed in vitro by SV40, media conditioned by 6 of 11 tumor-derived cell lines did not inhibit TU. Our results do not support the hypothesis that an antiproliferative factor secreted by SV40-transformed cells promotes the tumor-inducing capacity of these cells. Furthermore, inhibition of TU does not appear to be due to the production of a specific antimitotic peptide, but rather to other biochemical properties of the media conditioned by transformed cells. Finally, these biochemical properties do appear to correlate with specific morphological and growth characteristics of the tumor cells, but probably as an effect and not a cause. PMID- 2555052 TI - Identification and characterization of a Mr 50,000 adrenal protein in human hepatocellular carcinoma. AB - A Mr 50,000 cell surface protein antigen (p50) was identified on a human hepatocellular carcinoma derived cell line (FOCUS) by two monoclonal antibodies (SF 31 and SF 90). This antigen was subsequently shown to be expressed in vivo in human hepatocellular carcinoma. All 18 tumors tested by Western immunoblotting demonstrated high levels of p50 with undetectable amounts observed in the adjacent normal liver counterparts. Further characterization revealed that p50 is a monomeric polypeptide with a neutral pI (6.5-7.2) and appears not to be glycosylated. The cellular localization was determined by direct antibody binding to intact cells, immunoprecipitation of 125I-labeled cell surface proteins, and Western immunoblotting of subcellular fractions. p50 was found on the cell surface as well as in the cytoplasm. In vitro monoclonal antibody binding studies indicate that the protein is expressed in all human malignant cells (n = 34) tested thus far regardless of the embryonic tissue of origin and the degree of differentiation. p50 was present at very low levels in normal tissues with the notable exception of high expression in adrenal glands. The protein is conserved in mammalian evolution since a similar protein was also found in bovine adrenals. The molecular characteristics and the pattern of expression of p50 indicate that this normal adrenal protein is associated with the transformed phenotype. PMID- 2555054 TI - Identification and measurement of calcitonin precursors in serum of patients with malignant diseases. AB - Previous studies have suggested that molecular species larger than the mature calcitonin (CT) are produced by tumors of different origin. In order to study these species, we developed a monoclonal immunoradiometric assay for calcitonin precursors (CT-pr). This assay was based on both monoclonal antibody KC01 directed to the 1-11 region of katacalcin and monoclonal antibody CT08 directed to the 11-17 portion of CT. The sensitivity of this monoclonal immunoradiometric assay for CT-pr was less than 100 pg/ml. Only one of 131 healthy subjects had CT pr serum levels greater than 100 pg/ml; this value was therefore selected as the standard serum value in healthy individuals. CT-pr was present in the serum of seven of ten patients with advanced renal failure and in that of 21 of 52 patients (40%) with benign liver disease but was undetectable in sera of patients with other benign diseases. The serum CT-pr level was correlated with that of mature CT in patients with medullary carcinoma of the thyroid. In contrast, the serum CT-pr level was frequently elevated in the absence of a detectable CT level in patients with various malignant tumors and, particularly, in those with either tumors of the neuroendocrine system (60%) or hepatocellular carcinomas (62%). CT pr was detected in tumor extract from a patient with a hepatocellular carcinoma. Moreover, hybridization experiments with total RNA extracted from this tumor demonstrated the presence of RNAs hybridizing with complementary DNA encoding for common region, calcitonin, and katacalcin sequences. These results show that CT precursors are excreted by numerous cancers and might well be useful biological markers for the follow-up of productive tumors. PMID- 2555055 TI - Evidence for a mutant allele of the gene for DNA topoisomerase II in adriamycin resistant P388 murine leukemia cells. AB - Previous studies have shown that DNA topoisomerase II enzyme activity and protein levels are reduced in cloned lines of Adriamycin-resistant P388 leukemia cells relative to drug-sensitive cells (Deffie et al., Cancer Res., 49: 58-62, 1989). The molecular basis of the reduced topoisomerase II levels in these resistant cells has been investigated. Northern blot analysis of total cellular RNA from drug-sensitive and -resistant cells using a 1.8-kilobase human topoisomerase II complementary DNA revealed the presence of two mRNA species: a 6.6-kilobase transcript that was strongly expressed in drug-sensitive cells but reduced 7- to 8-fold in resistant cells; and a 5.5-kilobase transcript detected only in drug resistant cells. Southern blot analysis of genomic DNA digested with BamHI, StuI, or PvuII and probed with the 1.8-kilobase complementary DNA for human topoisomerase II showed that, in Adriamycin-resistant cells, there were two different alleles for topoisomerase II, one identical to the native allele but with a lower gene copy number than that found in sensitive cells, and a second allele containing a mutation present only in resistant cells. These findings suggest that the reduced levels of topo II protein in drug-resistant cells may be due to reduced amounts of the native 6.6-kilobase mRNA. The unique 5.5-kilobase mRNA in resistant cells may represent a shortened transcript of the mutated topoisomerase II allele. PMID- 2555056 TI - Synthesis and secretion of plasminogen activators and plasminogen activator inhibitors in cell lines of different groups of human lung tumors. AB - Several human lung tumor cell lines derived from large cell, squamous cell, and small cell carcinomas, as well as from mesotheliomas of the lung have been investigated for their gene expression and secretion of urokinase-type plasminogen activator (u-PA), tissue-type plasminogen activator (t-PA), and plasminogen activator inhibitors 1 and 2. All bronchogenic non-small cell carcinoma-derived cell lines studied could produce either plasminogen activators, their inhibitors, or both components, whereas in small cell lung carcinoma cell lines and cell lines derived from mesothelioma of the lung, no substantial amounts of any of these substances were synthesized. In detail, a large cell carcinoma-derived cell line, LCLC 97TM1, constitutively secreted large amounts of plasminogen activator. Northern blot analysis revealed RNA specific for u-PA and t-PA. Another large cell carcinoma-derived cell line, LCLC 103H, secreted smaller amounts of plasminogen activator and, additionally, plasminogen activator inhibitor. Specific mRNAs for u-PA and plasminogen activator inhibitors 1 and 2 were found in this cell line. In contrast, squamous cell carcinoma-derived cell lines secreted plasminogen activator only after treatment with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate; enhanced levels of u-PA, t-PA, and plasminogen activator inhibitor 1 mRNAs could then be demonstrated. The different expression of the plasminogen activator enzyme system distinguishes cell lines derived of non-small cell lung carcinoma from those of small cell lung carcinoma and may also reflect significant differences in the biological behavior of these tumor types. PMID- 2555058 TI - Characterization and distribution in normal and tumoral human tissues of breast cancer-associated antigen defined by monoclonal antibody 7B10. AB - Monoclonal antibody 7B10, raised against the human breast cancer cell line T47D, identifies an antigen found in human breast carcinomas and in normal breast. Western blot and immunoprecipitation studies detected a Mr 76,000 antigen in cytosol, cell membrane, and cell culture supernatants of T47D cells. 7B10 binding to T47D cell extracts was affected by proteolytic digestion with protease type VI, trypsin, and subtilisin while it was not altered by neuraminidase digestion. Adsorption of breast cancer cell line extracts with concanavalin A reduced 7B10 immunoreactivity more than 70%. These results suggest that the antigen is a glycoprotein and that the epitope does not contain sialic acid. 7B10 was reactive with neither human milk fat globule membrane, nor skimmed milk, nor the milk derived HBL 100 cell line. Conversely binding was detected in more than 50% of normal breast epithelial cells in culture. 7B10 immunostaining was positive on frozen sections of normal breast and nonmalignant mastopathies in 30 to 90% cells. In frozen sections of other normal tissues, 7B10 immunoreactivity was detected only in colon, apocrine glands of skin, parotid ducts, and luteal phase endometrium, confirming previous data on paraffin sections. Strong, homogeneous immunostaining was observed on frozen sections of intraductal and invasive lobular breast carcinomas (100% of cases), while more heterogeneous staining was found on invasive ductal carcinomas. Colon and rectal carcinomas, one carcinoma of the esophagus, and some cells in serous ovarian carcinomas also showed 7B10 reactivity. Immunoblotting of the 7B10-immunoreactive fraction isolated by Sepharose CL-6B chromatography of a breast carcinoma tissue sample extract identified the Mr 76,000 antigen, which was also detected in several breast cancer specimens, in colon adenocarcinomas, and in serous ovarian carcinoma fresh tumor extracts. The Mr 76,000 glycoprotein described here represents a breast cancer-associated antigen previously undescribed, mainly expressed in normal breast and breast tumors. PMID- 2555059 TI - Suppression of an epidermal growth factor receptor-hyperproducing tumor by an immunotoxin conjugate of gelonin and a monoclonal anti-epidermal growth factor receptor antibody. AB - An immunotoxin was made by conjugating a murine monoclonal antibody (B4G7) that recognizes the human epidermal growth factor (EGF) receptor with gelonin, a ribosome-inactivating protein. This B4G7-gelonin conjugate was shown to be specifically cytotoxic for EGF receptor-hyperproducing cells. The conjugate was tested in nude mice and shown to be capable of suppressing the growth of an EGF receptor-hyperproducing squamous carcinoma cell (A431) solid tumor. Nude mice bearing an A431 cell tumor that were given injections i.p. for 5 consecutive days with at least 10 micrograms of the conjugate showed significant suppression of tumor growth for about 7 days. On the other hand, an unconjugated mixture of B4G7 and gelonin showed no specific antitumor activity against the A431 cell tumor. The growth of an EGF receptor-deficient small cell lung cancer cell (H69) tumor was not suppressed by injection of the conjugate. No toxic effects were observed in histological examination of nontumorous tissues of mice treated with at least 250 micrograms of conjugate per mouse. These results suggest that the conjugate may be useful for targeting therapy to EGF receptor-hyperproducing squamous carcinoma. PMID- 2555057 TI - Prognostic value of receptors for insulin-like growth factor 1, somatostatin, and epidermal growth factor in human breast cancer. AB - The prognostic significance, as well as the relationship with known prognostic factors in breast cancer, of insulin-like growth factor 1 receptor (IGF-1-R), epidermal growth factor receptor (EGF-R), and somatostatin receptor (SS-R) was evaluated. IGF-1-R was positively correlated with estrogen receptor and age, but not significantly with progesterone receptor, lymph node status, and tumor size. EGF-R was negatively correlated to estrogen receptor and progesterone receptor, whereas no association was found with age, lymph node status, and tumor size. The levels of the tumor contents of IGF-1-R and EGF-R were not significantly related to tumor recurrence in 214 patients (test for trend, P = 0.20 and P = 0.08, respectively). However, patients with tumors containing intermediate levels of EGF-R (0.5 to 2.0 fmol/mg of membrane protein) experienced a longer disease-free survival than did patients with tumors possessing lower or higher levels of EGF R. This effect was most pronounced in the subgroup of patients with positive axillary lymph nodes: 66% disease-free after 5 yr compared with 38% and 46% for the groups with lower and higher EGF-R levels, respectively. The relapse-free survival for patients with tumors containing SS-R (15%) was significantly longer than for patients with SS-R-negative tumors (82% versus 46% disease free after 5 yr, P = 0.04). Assessment by multivariate analysis showed that lymph node status, tumor size, and differentiation grade were independent prognostic factors for relapse. In the Cox model, estrogen receptor and progesterone receptor were both negatively correlated with tumor recurrence, whereas overall EGF-R and IGF-1-R did not show such a relation. PMID- 2555060 TI - Effect of nerve growth factor on the transplacental induction of neurinomas by ethylnitrosourea in Sprague-Dawley rats. AB - Administration of nerve growth factor (NGF) to the offspring of Sprague-Dawley rats transplacentally exposed to 50 mg/kg ethylnitrosourea on the 20th day of gestation resulted in a significant reduction of trigeminal and peripheral nerve neurinomas. Forty, 60, and 80 micrograms of NGF was administered in five s.c. doses, one dose on each of days 12-16, 90-94, and 210-214 postnatally. Of the 34 rats in the NGF-treated group, 11 animals were affected with trigeminal nerve neurinomas as compared to 18/34 in the NGF-untreated group (P less than 0.05). In the peripheral nerves (spinal cord nerve roots) there were five and 11 neurinomas, respectively, in each group of 34 rats. When the total numbers of neurinomas (trigeminal and peripheral nerves) between these groups were compared (16/34 versus 29/34), the significance of neurinoma reduction was P less than 0.01. Five trigeminal and two peripheral neurinomas in the NGF-untreated group were shown by immunohistochemical staining to contain nerve growth factor receptor protein, whereas none of the neurinomas in the NGF-treated group were positive for the receptor protein. The results obtained from this experiment lend support to the hypothesis that NGF has the capability to reduce the oncogenic consequences of ethylnitrosourea exposure perhaps by the process of maturation and/or differentiation of the transformed cells, and that this effect may depend upon the presence of receptor binding sites. PMID- 2555061 TI - Growth and differentiation of a human T-cell leukemia cell line, CCRF-CEM, grafted in mice. AB - The growth of human CCRF-CEM T-cell lymphoblastic leukemia was studied in mice immune deprived by different techniques, and in CD-nu/nu athymic mice. Female CBA/CaJ mice were immune deprived by infant thymectomy, priming with 1-beta-D arabinofuranosylcytosine (200 mg/kg) 48 h prior to total body irradiation (925 cGy) designated theta ara-C gamma; or after thymectomy the mice received 925 cGy total body irradiation with marrow reconstitution (4 x 10(6) nucleated cells), designated theta gamma BM. Only in mice immune deprived by theta gamma BM, subsequently given a single dose of cyclophosphamide (100 mg/kg) 18-24 h before transplantation of CCRF-CEM, was there progressive reproducible engraftment and tumor growth. For mice immune deprived in this manner the tumor engraftment rate was 100 and 80% of tumors achieved greater than or equal to 1 cm3 within 46 days. In immune-deprived CBA/CaJ mice, but not CD-nu/nu athymic mice, tumor transplanted to the s.c. site metastasized to paraaortic and axillary nodes. Metastatic spread to lymph nodes was confirmed by immunophenotyping and by karyotyping. In contrast to the CCRF-CEM cells in culture, which expressed cytoplasmic CD3 (T3) but not surface CD3, both s.c. and metastatic CCRF-CEM line was exposed to phorbol-12-myristate 13-acetate in vitro to mimic the apparent differentiation which occurred in the xenografted cells, and a similar expression of surface CD3 after treatment was seen. This surface expression of CD3 was accompanied by production of mRNA for the T-cell receptor alpha chain and surface expression of the T-cell receptor. Identical T-cell receptor beta and gamma chain gene rearrangements were found for the CCRF-CEM line in vitro and the xenografted cells in vivo, demonstrating that only one clone was present and that differences in immunophenotyping were not the result of clonal selection. These results suggest that host (mouse) hematopoietic factors could affect human leukemic cell differentiation. PMID- 2555062 TI - Expression of functional neurotransmitter receptors in an uninnervated tissue: avian amnion. AB - The smooth muscle of the avian amnion is unusual because it is normally never innervated. However, as assessed by contractile response, this tissue expressed at least 11 different types of receptor for neurotransmitter substances including acetylcholine, norepinephrine, histamine, 5-hydroxytryptamine, vasoactive intestinal peptide, urotensin II, neurotensin, and somatostatin-28. Three neurotransmitters, histamine, 5-hydroxytryptamine, and norepinephrine, each acted via 2 separate and antagonistic types of receptors. The amnion also responded to prostaglandin E2. On the other hand, the tissue did not respond to substance P or bradykinin, 2 peptides that are known to affect smooth muscle contractility in a variety of other systems. Studies with organ-cultured amnion demonstrated that the smooth muscle can be cultured early in development and will differentiate in vitro. Some, but not all, of the amniotic responses developed in a defined medium. The results indicate that this novel smooth muscle preparation will be useful for identifying epigenetic factors that control the expression of functional receptors. PMID- 2555063 TI - Eradication of adenovirus E1-induced tumors by E1A-specific cytotoxic T lymphocytes. AB - Cytotoxic T lymphocyte (CTL) clones against adenovirus type 5 (Ad5) early region 1 (E1)-transformed cells were generated in C57BL/6 (B6) mice. A defined peptide encoded by Ad5 E1A is the target structure for H-2Db-restricted CTLs. Upon intravenous injection into B6 nude mice bearing Ad5 E1-induced tumors, these CTLs, if combined with recombinant IL-2, destroy subcutaneous tumor masses up to 10 cm3. The in vivo action of CTLs is highly specific, and long-term "memory" persists in treated nude mice months after tumor regression. Our data show an important role for CTLs directed against a viral nuclear oncogene product in tumor eradication. PMID- 2555064 TI - Positive and negative regulation of gene transcription by a retinoic acid-thyroid hormone receptor heterodimer. AB - We present evidence that the human thyroid hormone receptor forms a heterodimer with the human retinoic acid receptor. This interaction results in a cooperative increase in binding of the alpha retinoic acid receptor to a subset of thyroid hormone response elements. Mutations within the DNA binding domain or near the C terminus abolish either receptor's ability to interact cooperatively on these elements. The thyroid hormone-retinoic acid receptor heterodimer exhibits novel transcriptional properties in that coexpression of both receptors at low levels in Green monkey kidney (CV1) cells results in a positive transcriptional effect on promoters containing a palindromic thyroid hormone response element, but has a surprisingly negative effect on a thyroid hormone response element derived from the alpha myosin heavy chain gene. These results suggest that by forming heterodimers, more elab-orate control of transcription can be achieved by creating receptor combinations with differing activities. PMID- 2555065 TI - Expression of gelsolin by Cos cell secretion. PMID- 2555066 TI - Protein phosphorylation: the second messenger signal transducer of flagellar motility. PMID- 2555067 TI - [Tumors caused by HP viruses in children in various locations, multiplicity of the tumors and possibilities of modern therapy]. AB - The author gives an account of tumours caused by HP virus, its different antigenic types, the possibility of the incidence of different types of tumours at different sites in the same patient and the possibility of their transfer. The author gives also an account of hitherto used treatment of papillomatosis of the larynx in children and justification of immunocorrective treatment. PMID- 2555068 TI - [Verrucous carcinoma of the larynx]. AB - The authors give an account on the treatment and their own observations of the biological behaviour of verrucous carcinoma in five patients treated in the Radiotherapeutic Institute of the Municipal Institute of National Health and ENT Clinic of the Institute for Postgraduate Medical Training. They recommend in verrucous formations of the larynx bioptic examination and investigation of the presence of koilocytes. Koilocytes are generally considered a biological correlate of the presence of the HPV virus. At present the HUV virus, types 16, 18 and 30, are considered the possible aetiological agent of verrucous carcinoma. The method of choice in the treatment of verrucous carcinoma is surgery. PMID- 2555069 TI - Footprinting analysis of sequence-specific DNA-drug interactions. AB - Footprinting is a technique widely used in the analysis of sequence-specificity of proteins and drugs which bind to DNA in an equilibrium manner. Footprinting studies combine the enzymatic or chemical cleavage of DNA with the analysis of the resulting products by sequencing gels, thus revealing the position and length of individual binding sites. This review compares the information that can be obtained by using different footprinting agents and the characteristics of DNA drug interactions that can be inferred from footprinting results. PMID- 2555070 TI - Formation, resealing and persistence of DNA breaks produced by 4 demethoxydaunorubicin in P388 leukemia cells. AB - The formation and disappearance of DNA single-strand breaks (SSB) produced by 4 demethoxydaunorubicin (4-dmDR) in P388 murine leukemia cells and in a resistant subline were examined by alkaline elution methods in relation to cellular pharmacokinetics. DNA strand breaks produced by this intercalating agent were essentially DNA lesions mediated by topoisomerase II, even at very high drug concentrations, since they were detected as protein-associated breaks by filter elution. Similarly, the appearance of delayed DNA breaks in cells exposed to high concentrations, following drug removal, showed predominance of protein-associated breaks, thus supporting a similar mechanism of breakage induction. This finding indirectly suggests that, in this experimental model, free radical production makes little (if any) contribution to DNA damage, and also that DNA effects are not the consequence of early cell death. In contrast to a rapid disappearance of protein-associated strand breaks produced by intercalating agents and topoisomerase II inhibitors of different classes, DNA breaks induced by low concentrations of the anthracycline derivative are only partially reversible following drug removal, but they persisted and even increased with high concentrations. Thus, not only the extent of DNA breaks but also their persistence may contribute to the cytotoxic potency of anthracyclines. The importance of DNA lesions to cytotoxic action of the anthracycline is also emphasized by drug effect on the resistant line. A negligible effect on DNA of resistant cells was detected at drug concentrations lethal to sensitive cells. However, exposure to equitoxic drug concentrations resulted in a comparable amount of DNA breaks in sensitive and resistant cells. Although faster DNA rejoining in resistant cells may be in part attributable to increased efflux of drug, no correlation exists between cell drug accumulation and extent of DNA lesions. With equitoxic drug concentrations cellular drug content was higher in resistant cells, suggesting an intrinsic insensitivity of this variant to DNA cleavage effects of the anthracycline. PMID- 2555071 TI - Methylation of DNA by three N-nitroso compounds: evidence for sequence specific methylation by a common intermediate. AB - Methylation in vitro of calf thymus DNA, a supercoiled plasmid, poly(dG).poly(dC), and poly(dGdC).poly(dGdC) by N-nitroso(acetoxy methyl)methylamine and N-nitroso(acetoxybenzyl)methylamine in the presence of esterase, and by N-nitrosomethylurea was investigated. Although there were differences in the amounts of 7-methylguanine and O6-methylguanine formed in the various DNA substrates, the methylation pattern was the same for each of these methylating agents. The three compounds reacted identically when methylation of a portion of a 345 bp restriction fragment of the plasmid pBR322 was examined at nucleotide resolution by a sequencing assay. They also showed a tendency to react preferentially with particular guanines. These data suggest that the three N nitroso compounds methylate DNA via a common intermediate such as the methyl diazonium ion, which exhibits some sequence specificity. PMID- 2555073 TI - Binding of chlorinated benzidines to the rat hepatic aromatic hydrocarbon receptor. AB - The binding of benzidine, 3,3'-dichlorobenzidine (3,3'-Cl2BZ), and the asymmetrically-substituted chlorinated benzidines 3,5-dichlorobenzidine (3,5 Cl2BZ) and 3,5,3'-trichlorobenzidine (Cl3BZ) to the rat hepatic cytosolic aromatic hydrocarbon (Ah) receptor was measured, in order to assess the mechanism of P-450I induction by 3,3'-Cl2BZ. Cl3BZ is the most mutagenic benzidine derivative in the Ames assay. Binding affinity to the Ah receptor protein was determined by displacement of labelled 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) from the receptor, measured with the sucrose density gradient centrifugation technique. The rank order of affinities and the apparent inhibitor constants were: Cl3BZ (4 microM) greater than 3,5-Cl2BZ (8.4 microM) greater than 3,3' Cl2BZ (10 microM). Benzidine did not displace TCDD from the receptor protein. 4 Aminobiphenyl a structural link between the benzidine and biphenyl series competed weakly with TCDD. The 50% inhibition concentration was about 150 microM. The results are consistent with the hypothesis that the induction of P-450 enzymes by 3,3'-Cl2BZ in vivo is mediated by the Ah receptor. PMID- 2555072 TI - Metabolic activation of hydroquinone by macrophage peroxidase. AB - Lysates from macrophages, cells involved in hematopoiesis and immunological responses, catalyzed the metabolic activation of the benzene metabolite, hydroquinone, to protein-binding compounds and to free 1,4-benzoquinone. This reaction is mediated by a peroxidase since activation was dependent upon H2O2 and was prevented by the inhibitors aminotriazole and azide. Activation of hydroquinone was independent of HO. radicals since protein binding occurred in the presence of the HO. scavengers mannitol and dimethyl sulfoxide. In reactions with macrophage lysates, phenol, another hepatic metabolite of benzene, stimulated the production of 1,4-benzoquinone as well as the amount of hydroquinone equivalents bound to protein in a dose-dependent manner. Addition of cysteine to incubations with macrophage lysates resulted in a dose-dependent decrease in hydroquinone equivalents bound to protein. At 100 microM cysteine, protein binding was inhibited by 63% and this decrease was recovered as the monocysteine-hydroquinone conjugate. Macrophages catalyzed the arachidonic acid mediated activation of hydroquinone to metabolites which bound to cellular macromolecules. This activation was inhibited by indomethacin indicating the action of prostaglandin synthase in hydroquinone metabolism by macrophages. The results of these experiments demonstrate that macrophage peroxidase catalyzes the metabolic oxidation of hydroquinone to 1,4-benzoquinone and that 1,4-benzoquinone and/or its semiquinone intermediate are binding to protein and cysteine. Hydroquinone activation by macrophages and subsequent macromolecular binding may be associated with the immunologic and hematopoietic toxicity of benzene. PMID- 2555074 TI - [Establishment of immunohistochemical technique for polyamine and its application in research of lung cancer]. AB - Antiputrescine, antispermidine and antispermine were used to detect polyamine in normal and cancer tissues of lung by PAP method. Sections of lung cancer, which were known to have high concentration of polyamine, were used as positive control. Results showed highly significant difference in the staining between the experimental and control groups. Immunohistochemical assay further demonstrated that polyamine located mainly in the nucleus, nucleolus and cytoplasm of the cancer cells and was evenly distributed. The staining results of normal lung tissue and epithelium of bronchi were negative or weakly positive. The morphological finding of this experiment is considered important for further study on the correlation of polyamine with the differentiation, proliferation and malignant change of cells. PMID- 2555075 TI - [Relationship between malignant transformation of ovarian mucinous cystadenoma and intestinal epithelial metaplasia]. AB - Ninety cases of ovarian mucinous tumors were studied histologically and histochemically. Intestinal metaplasia was found in 48.2% (14/29) of benign. 73.6% (14/19) of borderline and 92.9% (39/42) of malignant mucinous cystadenomas. The differences between these three groups are statistically significant (P less than 0.01). Among 67 cases of intestinal metaplastic mucinous tumors, 43 contained argyrophil cells, and 36 contained argentaffin cells. The coexistence of intestinal metaplasic and uterocervical canal type epithelia was observed in 2/3 of borderline and 1/3 of malignant intestinal mucinous cystadenomas. In addition, there were 5 cases of borderline and 3 cases of malignant uterocervical canal type mucinous cystadenomas among the 90 cases. It is evident that the malignant transformation of ovarian mucinous cystadenoma was closely related to intestinal metaplasia. Anyhow, it seems not necessary for malignant transformation of all ovarian mucinous cystadenomas to pass through a stage of intestinal metaplasia: some of the malignant mucinous cystadenomas were considered probably originating from the uterocervical canal type epithelium. PMID- 2555076 TI - Comparison of endocrinological stress response associated with transvaginal ultrasound-guided oocyte pick-up under halothane anaesthesia and neuroleptanaesthesia. AB - Twelve patients with mechanical infertility in the in vitro fertilization program were studied. Seven of them received halothane anaesthesia and the other five received neuroleptanaesthesia. Higher plasma prolactin levels and lower plasma progesterone levels were observed in the neuroleptanaesthesia group than in the halothane group during and after transvaginal ultrasound-guided oocyte pick-up. Plasma adrenocorticotropic hormone and cortisol levels of the patients suggested that surgical stress was minimal in both groups. It is likely that droperidol and fentanyl, both used in neuroleptanaesthesia, were responsible for the hyperprolactinaemia which was followed by inhibition of progesterone production. These agents, therefore, are not recommended as anaesthetic agents for transvaginal ultrasound-guided oocyte pick-up. PMID- 2555077 TI - Endothelium-dependent relaxation and cyclic GMP accumulation in rabbit pulmonary artery are selectively impaired by moderate hypoxia. AB - The effect of hypoxia on endothelium-dependent and endothelium-independent vasodilation was studied in phenylephrine-precontracted, isolated rings of rabbit first-branch pulmonary artery. Concentration-dependent relaxation responses to the endothelium-dependent dilators methacholine, ATP, and the calcium ionophore (A23187) as well as to the endothelium-independent dilators sodium nitroprusside and isoproterenol were obtained before, during, and after exposure to hypoxia (PO2 = 42 +/- 1 mm Hg) in the presence of indomethacin (2.8 x 10(-5) M). This moderate degree of hypoxia inhibited (p less than 0.05) endothelium-dependent but not endothelium-independent relaxation responses without producing irreversible vascular damage. In parallel experiments, cyclic GMP accumulation in pulmonary vascular rings in response to maximal doses of the above vasodilators was measured in the presence and absence of hypoxia. Cyclic GMP accumulation in response to endothelium-dependent dilators (methacholine, ATP, and A23187) was inhibited (p less than 0.05) by hypoxia while cyclic GMP accumulation in response to the endothelium-independent dilator sodium nitroprusside was not. When phenylephrine precontracted vessels were exposed to hypoxia in the absence of vasodilators, a small, transient increase in tension occurred, which was greater in endothelium-intact than in endothelium-denuded vessels (0.70 +/- 0.12 vs. 0.09 +/- 0.03 g, respectively; p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555078 TI - Beneficial effects of alpha 2-adrenoceptor activity on ischemic myocardium during coronary hypoperfusion in dogs. AB - We have previously reported that alpha 2-adrenoceptor stimulation enhances adenosine-induced coronary vasodilation. In the present study, we tested the hypothesis that alpha 2-adrenoceptor activity exerts beneficial effects on myocardial ischemia through augmentation of vasodilatory effects of released adenosine. In open-chest dogs, the left anterior descending coronary artery was perfused through an extracorporeal bypass tube from the carotid artery. Propranolol was infused into the bypass tube to exclude the metabolic effects of norepinephrine. When clonidine (0.24 micrograms/kg/min i.c.) was infused for 10 minutes after reduction of coronary blood flow by partial occlusion of the bypass tube, coronary blood flow was increased by 43% from 27 +/- 1 ml/100 g/min despite no changes in coronary perfusion pressure (38 +/- 5 mm Hg) and a slight decrease in adenosine release. Both fractional shortening and lactate extraction ratio of the perfused area were significantly improved (fractional shortening, 1.8 +/- 1.0 to 10.9 +/- 1.5%, p less than 0.001; lactate extraction ratio, -57.8 +/- 6.5 to 31.9 +/- 2.4%, p less than 0.005). Identical results were observed in the denervated hearts, indicating that the beneficial effect of clonidine is not attributed to the prevention of norepinephrine release from the sympathetic nerve terminals. The beneficial effects of clonidine were prevented by yohimbine, an alpha 2-adrenoceptor blocking agent. An adenosine receptor antagonist, 8 phenyltheophylline, also prevented the beneficial effects of clonidine, indicating that these beneficial effects are mediated by effects of adenosine. Furthermore, the extent of augmentation of coronary flow in the ischemic heart was coincided with that of augmentation of exogenous adenosine-induced hyperemic flow (40%) by clonidine. Production of cyclic AMP in the coronary artery during myocardial ischemia was augmented by clonidine. In 12 other dogs, myocardial ischemia was produced by intracoronary embolization of microspheres (15 microns in diameter). Clonidine enhanced (39%) the hyperemic coronary flow and improved both fractional shortening and lactate extraction ratio. Thus, we conclude that alpha 2-adrenoceptor stimulation can ameliorate myocardial ischemia mainly due to enhancement of vasodilatory effects of adenosine released from the ischemic myocardium. PMID- 2555079 TI - The role of alpha 1- and alpha 2-adrenergic receptors in mediation of coronary vasoconstriction in hypoperfused ischemic myocardium during exercise. AB - This study was carried out to test the hypothesis that adrenergic coronary vasoconstriction limits blood flow to hypoperfused regions of myocardium during exercise. The vasoconstrictor influence of alpha-adrenergic receptor subtypes was assessed by use of selective adrenergic blocking agents. Dogs chronically instrumented with a circumflex coronary artery hydraulic occluder and an intra arterial catheter underwent treadmill exercise in the presence of a coronary stenosis that decreased distal perfusion pressure to 40 mm Hg. Myocardial blood flow was measured with radioactive microspheres (15 microns) before and during selective alpha 1- or alpha 2-adrenergic receptor blockade produced by intracoronary infusion of prazosin (1 microgram/kg/min x 10 min) or idazoxan (1 microgram/kg/min x 10 min), respectively. Coronary perfusion pressure was held equal before and during receptor blockade with the hydraulic occluder. Compared with control exercise, subendocardial blood flow increased during alpha 1 receptor blockade with prazosin from 0.60 +/- 0.14 to 1.12 +/- 0.17 ml/min/g (p less than 0.05), and mean transmural flow increased from 1.07 +/- 0.19 to 1.60 +/ 0.22 ml/min/g (p less than 0.05). In contrast, subendocardial and mean transmural blood flow were not different from control during selective alpha 2 adrenergic receptor blockade with idazoxan (0.48 +/- 0.10 vs. 0.67 +/- 0.14 ml/min/g, p = 0.33, and 0.82 +/- 0.15 vs. 1.02 +/- 0.20 ml/min/g, p = 0.45, respectively). These data indicate that even in the presence of a coronary stenosis that causes substantial myocardial underperfusion during exercise, residual coronary vasoconstrictor tone is present in ischemic myocardium, and this vasoconstriction is mediated predominantly by the alpha 1-adrenergic receptor. PMID- 2555080 TI - Multiple types of Ca2+ currents in single canine Purkinje cells. AB - Whole-cell Ca2+ channel currents were recorded from isolated single canine Purkinje and ventricular cells to determine whether there were multiple types of Ca2+ channels in these two cell types, as in many other excitable tissues. The experimental conditions were such that currents other than Ca2+ channel currents were largely suppressed. The charge carrier was either Ca2+ or Ba2+ (5mM). In every canine Purkinje cell studied (n = 36), we saw T and L Ca2+ channel currents that are similar to their counterparts in other tissues. Neither current was affected by tetrodotoxin (30 microM), but both were reduced by Mn2+ (5mM). Ni2+ (50 microM) blocked T more than L current. Nisoldipine (1 microM) apparently abolished the L current but also decreased the T current by 50%. Substitution of Ba2+ for Ca2+ augmented and prolonged L current but did not affect T current significantly. At 36 degrees C and with 5 mM [Ca2+]o, T current inactivated over a voltage range from -70 to -30 mV whereas L current inactivated between -30 and +20 mV. T current was detectable in only some of the ventricular cells studied (8 out of 12). In these cells the ratio of maximal T current to maximal L current (0.2 +/- 0.1, n = 8) was lower than the T/L ratio in Purkinje cells (0.6 +/- 0.2, n = 6). The density of peak L current in ventricular cells (7.5 +/- 1.7 pA/pF, n = 8) was higher than that in Purkinje cells (4.4 +/- 3.4 pA/pF, n = 6). Therefore, in ventricular cells the L current is the main Ca2+ current whereas in Purkinje cells, the T current also contributes significantly to membrane electrical activity. In Purkinje cells, beta-adrenoceptor stimulation by isoproterenol (1 microM) increased L current but did not affect T current. On the other hand, in 70% (7 out of 10) of the Purkinje cells, alpha-adrenoceptor stimulation by 10 microM norepinephrine (in the presence of 2 microM propranolol) increased the T current. Our observations show that the distribution of the two types of Ca2+ channels in canine ventricle is heterogeneous and that the two types of Ca2+ channels are modulated by catecholamines by different receptors. PMID- 2555081 TI - Developmental changes in guanine nucleotide regulatory proteins in the rat myocardial alpha 1-adrenergic receptor complex. AB - During development, the cardiac alpha 1-adrenergic chronotropic response changes from positive in the neonate to negative in the adult. The negative chronotropic effect of alpha 1-adrenergic stimulation in the adult depends on maturation of sympathetic innervation and the presence of a pertussis toxin (PT)-sensitive guanine nucleotide-binding (G) protein. To examine the possibility of a developmental change in coupling of a PT-sensitive G protein to the alpha 1 adrenergic receptor, radioligand binding experiments with the iodinated alpha 1 selective radioligand [125I]-I-2-[beta-(4 hydroxyphenyl)ethylaminomethyl]tetralone ([ 125I]-IBE 2254) were performed on membranes prepared from control and PT-treated neonatal and adult rat hearts. Scatchard analysis showed fewer alpha 1-adrenergic receptors in the adult than in the neonate (168 +/- 10 fmol/mg protein in the neonate vs. 124 +/- 13 fmol/mg protein in the adult), but similar affinities (equilibrium dissociation constant 124 +/- 29 pM in the neonate vs. 140 +/- 34 pM in the adult). PT treatment did not alter the results. In both the neonate and adult, 5'-guanylylimidodiphosphate [Gpp(NH)p, 500 microM] shifted the l-epinephrine competition curve to the right and increased the slope factor toward unity. PT had no effect on the l epinephrine competition curve in the neonate. However, in the adult PT itself caused a partial shift in the agonist competition curve, reducing but not eliminating the effect of Gpp(NH)p. Consistent with the results from the binding experiments, PT did not have any effect on the alpha 1-adrenergic-mediated positive chronotropic response in the neonate, whereas in the adult the alpha 1 adrenergic-mediated negative chronotropic response was completely converted to a positive one after PT treatment. These results indicate the presence of a PT insensitive G protein in the neonatal and adult rat heart and the acquisition of a PT-sensitive G protein linked to the negative chronotropic response during development. PMID- 2555082 TI - EDRF increases cyclic GMP in platelets during passage through the coronary vascular bed. AB - It was investigated whether endothelium-derived relaxing factor (EDRF) increases cyclic GMP (cGMP) content in platelets passing through the coronary bed. Boluses of washed platelets from healthy human donors were injected into the aortic perfusion line of isolated, saline-perfused rabbit hearts under constant flow conditions (28 +/- 2 ml/min). The coronary effluent was collected over 5 seconds, and the cGMP content of platelets was determined by radioimmunoassay. Platelet cGMP amounted to 0.34 +/- 0.11 pmol/mg protein after passage through the unstimulated coronary bed. During stimulation with acetylcholine (1 microM), it increased to 1.6 +/- 0.5 pmol/mg (p less than 0.01; n = 14). Simultaneously, the platelet recovery (measured over 20 seconds after injection) was enhanced (by 45 +/- 11%; p less than 0.01) during endothelial stimulation with acetylcholine. Treatment with the EDRF inhibitor hemoglobin (6 microM) completely abolished the increase in platelet cGMP (p less than 0.01; n = 11) as well as the enhanced platelet recovery (n = 8). Inhibition of EDRF by hemoglobin reduced also the basal platelet cGMP content to 0.17 +/- 0.11 pmol/mg (p less than 0.01). The data indicate that basally released EDRF is able to increase cGMP in platelets during a single passage through the coronary bed. The enhanced recovery of platelets after EDRF stimulation, which coincides with an increase of platelet cGMP, suggests that EDRF plays an important role as inhibitor of platelet activation in the coronary circulation. PMID- 2555084 TI - Simple enzymatic determination of total cholesterol in gallstones. AB - In this simple method for rapid enzymatic determination of total cholesterol in human gallstones, gallstone powder is dissolved in an 80/20 by vol mixture of N,N dimethylformamide and dimethyl sulfoxide and reacted directly with the aqueous enzymatic reagent, without further treatment. The organic solvents do not interfere with enzymatic activities of cholesterol oxidase, esterase, and peroxidase in the assay mixture. The method is reproducible (CVs 3.7% to 6.6%), analytical recoveries ranged from 98.6% to 102%, and linearity is good. Furthermore, results correlated well with those obtained by infrared spectroscopic measurements. PMID- 2555083 TI - Angiotensin II modulates cardiac Na+ channels in neonatal rat. AB - Since chronic congestive heart failure syndromes are associated with both elevated circulating levels of angiotensin II and potentially lethal ventricular tachyarrhythmias, we investigated the effect of angiotensin II on voltage dependent cardiac Na+ currents. Single-channel Na+ currents in neonatal rat ventricular myocytes were studied using the patch clamp method in the cell attached mode. Angiotensin II applied outside the patch increased the frequency of opening and rates of activation and inactivation of single-channel Na+ currents within the patch. These effects were mimicked by the phorbol ester 12-O tetradecanoylphorbol-13-acetate (TPA) and were prevented by prior incubation with TPA. Therefore, we propose that angiotensin II modulates cardiac Na+ currents by a cytoplasmic second messenger, perhaps protein kinase C, and this may predispose toward arrhythmia. PMID- 2555085 TI - Comparison of agarose gel electrophoresis and a chaotropic method for lactate dehydrogenase isoenzyme-1. PMID- 2555086 TI - [Morphological changes in human diaphragm--ragged red fiber, core/targetoid fiber, cytoplasmic body, and ring fiber]. AB - We have already described that ragged red fiber (RRF), core/targetoid fiber and type 1 fiber predominance were found at autopsy in the diaphragm taken from patients with chronic obstructive pulmonary diseases. The purpose of the present study is to investigate morphological and histochemical changes in the diaphragm in denervating neurologic disorders. The diaphragm in the costal portion was taken from 22 autopsy cases including 4 with amyotrophic lateral sclerosis (ALS), 4 cerebrovascular diseases, 3 Parkinson disease, 2 olivopontocerebellar atrophy. In addition, 4 diaphragm muscles were biopsied at the time of surgery for lung cancer. In the diaphragm we observed not only RRF and core/targetoid fiber but also cytoplasmic body and ring fiber in many cases. These findings were, however, not specific for neurologic disorders. Focal cytochrome c oxidase deficiency was found in muscles with RRF. It should be emphasized that RRF was absent in 3 of 4 cases with ALS and in a case with elevated hemidiaphragm from phrenic nerve paralysis. In the previous report, we suggested that RRF was formed under the relative ischemic state in overworking diaphragm. The relative ischemia means a condition that oxygen (energy) demand for respiratory work exceeds over oxygen supply from the blood in the overworking diaphragm. The reason why no RRF was found in the denervated muscle is that the ischemic state in the denervated muscles is relieved by immobilization after denervation. Karpati et al conformed that denervation prevented ischemic state in the muscle. Other histochemical features in the diaphragm included cytoplasmic body and ring fiber.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555087 TI - ["Drop hand" resulting from cerebral infarction--case report]. AB - A 52-year-old male was admitted with the progressive weakness of the right 3rd, 4th and 5th fingers on three days after the onset. On admission, his right hand and all of right fingers were flexed. Any extension of the right hand and any movement of right fingers were not accomplished. The sensory deficit in the right wrist joint was not observed. Initial computed tomography (CT) scan of the brain carried out on the day of admission could not detect any abnormal findings. However, the follow-up CT scans demonstrated the localized low density lesion in the left frontoparietal area. MR imaging also revealed abnormality attributable to cerebral infarction. Diagnosis of "Pseudoradial nerve palsy" was made. The mechanisms of these neurological deficits and the problems in differential diagnosis between central and peripheral nerve involvements were discussed. PMID- 2555088 TI - Hereditary cerebral hemorrhage with amyloidosis--Dutch type. Research-Group Hereditary Cerebral Amyloid-Angiopathy. PMID- 2555089 TI - Cerebral toxoplasmosis in the acquired immune deficiency syndrome. AB - We examined the clinical and pathological features of 26 patients who presented with a diagnosis of CNS toxoplasmosis. Patient data was analyzed with respect to demographics, clinical presentation, treatment course and pathology. Patients presented with a wide variety of signs and symptoms. All patients had positive serum antitoxoplasma IgG; ring enhancing lesion(s) were present on all but one brain CT scans. A series of guidelines in the management of CNS toxoplasmosis in AIDS patients are presented. Prior to biopsy, patients with positive serology and characteristic CT scans should receive two weeks of treatment. Biopsy is indicated in those cases with negative serology, atypical presentation, progressive clinical deterioration, or differential response of lesions to empiric therapy. PMID- 2555090 TI - Clinical and pharmacokinetic evaluation of controlled-release levodopa/carbidopa (CR-4) in parkinsonian patients with severe motor fluctuations: a six month follow-up study. AB - The results of a six month open-label study comparing the efficacy of controlled release levodopa-carbidopa (Sinemet CR-4 200 mg/50 mg) with standard levodopa/carbidopa (250 mg/25 mg) in 17 patients with idiopathic Parkinson's disease and severe response fluctuations, are reported. Major clinical benefits included; improvement of disability, reduction of the number of 'off' periods (predominantly end-of-dose hypokinesia) and a slight increase in 'on' time. No improvement was observed in two of our patients. Mean levodopa plasma levels were comparable between the two types of formulations during optimal treatment, however systemic bioavailability was significantly higher with CR-4. Delayed onset of antiparkinsonian effect of CR-4, resulting from an increase of Tmax for levodopa, was one of the major complaints and required additional small amounts of standard levodopa in three patients. PMID- 2555091 TI - Merkel cell carcinoma. Long term survival in a patient with proven brain metastasis and presumed choroid metastasis. AB - Merkel cell carcinoma (Trabecular carcinoma) is a rare malignant tumor of the skin. Because these tumors tend to spread locally via the lymphatic system, very few papers have dealt with the treatment of distant metastasis. Systemic disease is uncommon and usually fatal. In this paper, we report a case of Merkel cell carcinoma with proven brain metastasis and a solid choroidal tumor. The patient responded well to radiation and chemotherapy and is alive and neurologically intact three years after diagnosis. All previous patients with metastatic Merkel cell carcinoma to the brain died within two months of the diagnosis. We use this case to discuss possible routes of metastatic dissemination and to review the treatment options in patients with distant metastatic disease. To our knowledge, this is the first reported case of presumed choroidal metastasis of Merkel cell carcinoma and the longest documented survival in a patient with brain involvement. PMID- 2555092 TI - Lipoma of the sylvian region. AB - An exceptional case of lipoma of the sylvian region is described and the other seven reported cases are reviewed. The sylvian fissure is the most rare site of intracranial lipomas. Sylvian lipomas may be asymptomatic or present with epileptic seizures due to irritation of the cortex of the sylvian fissure. Only two previous cases have been diagnosed during life by computerized tomography and operated on; the deep location of these lipomas and their adherences to the sylvian cortex and the branches of the middle cerebral artery make radical removal impossible and dangerous. PMID- 2555093 TI - Loss of motivation for speaking with bilateral lacunes in the anterior limb of the internal capsule. AB - A 76-year-old man suddenly presented with loss of motivation for speaking. He had no aphasia, depression or dementia, but he did not want to speak with other people. CT scan showed hypodensities, presumably lacunes, mainly in the anterior limb of the two internal capsules. Global behavioral changes with loss of self psychic activation have been reported, but in our patient the disturbance was restricted to oral language. The role of activating cortico-subcortical loops is discussed. PMID- 2555094 TI - Paraneoplastic subacute cerebellar degeneration in Hodgkin's disease. Report of three cases and review of the literature. AB - 12 cases of paraneoplastic cerebellar degeneration with Hodgkin's disease have so far been reported. Three additional patients (Stage II-IA/IIA/IIIB) are presented. Although CT and MR-imaging (MRI) revealed a marked cortical atrophy of the cerebellum, there was no evidence of direct brain involvement. After irradiation and chemotherapy all patients showed a complete remission. There was no evidence of primary disease after an observation period of 3/2/5 years. Cerebellar symptoms also showed a partial remission. As compared to the literature, our treatment appears to be more favorable, probably due to the fact, that therapy was more aggressive than usual. PMID- 2555095 TI - Intramedullary cysticercosis. AB - We are reporting two cases of intramedullary cysticercus cysts. Both patients presented with a clinical picture of progressive cord compression. One of them also had bilateral radicular pain. In the female patient the symptoms became exaggerated during her two consecutive pregnancies which is a rare and interesting observation. Excision of cyst resulted in significant improvement in both the patients. PMID- 2555096 TI - Giant sacral perineurial cyst. A case report. AB - A rare case of giant sacral perineurial cyst, causing sciatic pain, explored by myelography and computerized tomography, is reported. The cyst, associated with large erosion of the sacrum, was poorly visualized on the myelography, because of its large size, whereas it was better defined on CT scan. Sacral perineurial cysts are usually small and asymptomatic and rarely cause radicular symptoms. The radiological diagnosis and the treatment of these nerve root cysts are briefly discussed. PMID- 2555098 TI - Anterolateral lumbar meningocele presenting as an ovarian cyst, in a patient with neurofibromatosis. AB - A case of an anterolateral lumbar meningocele in a patient with neurofibromatosis is presented. As anterolateral lumbar meningocele may present itself only by non specific symptoms like low back pain or abdominal pain, such symptoms require prompt diagnostic work up in patients with neurofibromatosis. The significance of MR imaging in early recognition of this rare anomaly is emphasized. PMID- 2555097 TI - Chronic spinal arachnoiditis following intracranial subarachnoid haemorrhage. AB - A 54-year-old woman presented with progressive gait imbalance and increased urinary frequency, associated with spinal arachnoiditis. The symptoms started after the occurrence of communicating hydrocephalus as a sequel of subarachnoid haemorrhage (SAH), and were initially attributed to post-SAH vasospasm, decompensating hydrocephalus and/or periventricular leuko-encephalopathy. Further clinical deterioration led to the diagnosis of thoracic spinal arachnoiditis, as a second complication of SAH. PMID- 2555099 TI - Leptomeningeal plasmacytosis. Case report and considerations on treatment. AB - We describe a 68-year-old patient with a plasma cell leukaemia in haematological remission presenting with massive intracranial leptomeningeal plasmocytic infiltration (LPI) and hydrocephalus. He was treated with skull irradiation and a combination of intraventricular and lumbar intrathecal therapy with methotrexate. Neurologic improvement and clearance of plasma cells from the cerebrospinal fluid was reached after 2 weeks of treatment but prolonged follow-up was interrupted by a lethal gastro-intestinal haemorrhage, 6 weeks after starting the therapy. From previously reported cases it is known that LPI almost always occurs in either high-grade plasmocytomas or plasma cell leukaemia. These data suggest that therapy of LPI should be the same as in other leukaemias with leptomeningeal infiltration. PMID- 2555100 TI - Scientific meeting of The Netherlands Society of Child Neurology. Heeze, 7th of April 1989. Abstracts. PMID- 2555101 TI - Scientific meeting of the Flemish Society of Neuro-Psychiatry. Kortrijk, 15th April 1989. Abstracts. PMID- 2555102 TI - Scientific meeting of The Netherlands Society of Neurology. Amsterdam, 22nd of April 1989. Abstracts. PMID- 2555103 TI - Highly preferred targets for retrovirus integration. PMID- 2555104 TI - Protoporphyrin IX-induced impairment of biliary lipid secretion in the rat. AB - 1. In order to gain information on the effect of protoporphyrin IX on changes in the properties of the canalicular plasma membrane, we studied the release of canalicular membrane constituents, namely phospholipids, cholesterol and 5' nucleotidase, into bile in anaesthetized rats receiving saline or taurocholate (0.5 mumol min-1 100 g-1 body weight) with or without protoporphyrin IX infusion (10 or 20 micrograms min-1 100 g-1 body weight). 2. Protoporphyrin IX induced an impairment of spontaneous bile flow and of biliary secretion of cholesterol, phospholipids and bile acids. The taurocholate-induced increase in bile acid output was not significantly reduced by protoporphyrin IX at either of the doses used. However, when a cholestatic dose of protoporphyrin IX was infused, the taurocholate-induced bile flow and secretion of lecithin and cholesterol were significantly reduced. 3. Biliary output of phospholipid species other than lecithin did not counterbalance the protoporphyrin IX-induced reduction in biliary lecithin secretion. Biliary outputs of both total phospholipid and lecithin were inhibited by protoporphyrin IX to similar extents. 4. Protoporphyrin IX alone had no effect on the biliary release of 5'-nucleotidase, whereas when it was given with taurocholate, it increased the bile acid-induced biliary output of this enzyme markedly. 5. In summary, these results indicate that protoporphyrin IX impairs the biliary secretion of phospholipids and cholesterol but not that of bile acid. The release of canalicular membrane constituents other than lipids was also modified by protoporphyrin IX. PMID- 2555105 TI - Muscle enzyme adaptation in patients with peripheral arterial insufficiency: spontaneous adaptation, effect of different treatments and consequences on walking performance. AB - 1. The activities of phosphofructokinase (PFK), citrate synthetase (CS), lactate dehydrogenase (LDH), 3-hydroxyacyl-CoA dehydrogenase (ACDH) and cytochrome-c oxidase(Cyt-ox) in the calf muscle tissue were compared in subjects with intermittent claudication (n = 38) and controls (n = 20). The activities of CS, ACDH and Cyt-ox were increased and the activity of Cytox was positively correlated to the maximal walking distance (MWD) in the patients. 2. Thirty-three patients with intermittent claudication were randomized to three treatment groups: (1) operative surgery, (2) operative surgery supplemented with physical training and (3) physical training alone. Before and after 6-12 months of treatment, symptom-free walking distance (SFWD), MWD, ankle-brachial blood pressure quotient (ankle index), maximal plethysmographic calf blood flow (MPBF) and the activities of PFK, CS, LDH, ACDH and Cyt-ox were measured. 3. SFWD and MWD increased in all three groups. Ankle index and MPBF increased in groups 1 and 2, but were unchanged in group 3. The activities of Cyt-ox and CS decreased with operation, but the activity of Cyt-ox was further augmented with training in group 3. Overall, the change in ankle index explained 80-90% of the variability in walking performance. In a separate analysis, the increased activity of Cyt-ox in group 3 was positively correlated to, and explained 31% of the variability in, the improvement in SFWD. 4. These findings indicate that both physical activity and a reduced calf blood flow are necessary conditions for the enzymatic adaptation to take place. A causal relationship between metabolic adaptation in the muscle tissue and walking performance is suggested.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555106 TI - Endogenous digoxin-like immunoreactive factor and digitalis-like factor associated with the hypertension of patients receiving multiple alkylating agents as part of autologous bone marrow transplantation. AB - 1. Hypertension is a complication of autologous bone marrow transplantation when therapy includes multiple alkylating agents. We have sought to identify the factors underlying this hypertension. We measured weight, serum creatinine, plasma renin activity, aldosterone and digoxin-like immunoreactive factor (DLIF), by digoxin radioimmunoassay, in 18 patients. Plasma catecholamines were also measured in five patients. 2. Of the 18 patients studied, 15 became hypertensive. The variable most consistently associated with these individuals' hypertension was DLIF activity which was increased in 14 of the 15 hypertensive patients (P = 0.055, Fisher exact test). Serum creatinine was increased at some point in seven of the 15 hypertensive patients, weight was increased in five and plasma renin activity and aldosterone were increased in one. Catecholamines were not increased in any of the five patients in which they were measured. 3. The association between changes in mean arterial pressure (MAP) and changes in DLIF for the group as a whole was assessed by analysing one data pair per patient, representing the maximal MAP. This correlation was significant (r = 0.75, P = 0.001). 4. Within individual patients, changes in MAP and changes in serum DLIF concentrations were significantly correlated (r greater than 0.50, P less than 0.05) in six of 15 hypertensive patients. 5. Digitalis-like factor (DLF) was measured by inhibition of (Na+,K+)-adenosine 5'-triphosphatase in five patients and DLF and DLIF were significantly correlated (r = 0.081, P = 0.0001). DLF and MAP were also significantly correlated (r = 0.59, P = 0.002). 6. This represents the first longitudinal study of the relationship between DLIF and blood pressure in hypertensive individuals, and the results suggest that DLIF may contribute to the increased blood pressure in some of these subjects. PMID- 2555107 TI - Effects of endothelin on systemic and renal haemodynamics and neuroendocrine hormones in conscious dogs. AB - 1. The effects of endothelin on systemic and renal haemodynamics and plasma concentrations of neuroendocrine hormones including plasma renin activity, aldosterone, adrenocorticotropic hormone, cortisol, catecholamines and arginine vasopressin were investigated in 18 conscious dogs. 2. Bolus injection of 4 pmol of endothelin/kg did not cause any significant changes in haemodynamics. Mean arterial pressure was elevated by both doses of 40 pmol/kg [91 +/- 2 to 99 +/- 2 mmHg (12.1 +/- 0.3 to 13.2 +/- 0.3 kPa), P less than 0.05] or 200 pmol/kg [93 +/- 2 to 107 +/- 3 mmHg (12.4 +/- 0.3 to 14.3 +/- 0.4 kPa), P less than 0.01], the latter dose increasing cardiac output (14%, P less than 0.05) and heart rate (9%, P less than 0.05), and the former reducing these parameters (14% and 8%, P less than 0.05, respectively). 3. In contrast with the various changes in systemic haemodynamics, renal blood flow transiently increased immediately after bolus injection in a dose-dependent manner (28%, P less than 0.05, 50%, P less than 0.01 and 110%, P less than 0.01 with 4, 40 and 200 pmol of endothelin/kg, respectively). This transient elevation of renal blood flow was followed by a gradual decrease (16%, P less than 0.05; 31%, P less than 0.01 and 36%, P less than 0.01) at 10 min. 4. All neurohormones were elevated in a dose-dependent manner.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555108 TI - In vitro comparison between four variants of Aujeszky's disease virus. AB - Four Aujeszky's disease (pseudorabies) virus variants were characterized in vitro by investigation of their resistance to heat at 48 degrees C, sensitivity to trypsin and ability to replicate in pig alveolar macrophages, two of these variants (Ls-1 and Ls-2) were cloned previously from a single isolate of virus and showed differing pathogenicity for pigs; the virulent Stanley strain; and the non-virulent NIA-4 strain were included for comparison. Heat treatment produced slight decreases in infectivity but no significant differences were observed in the rates of inactivation. Both Ls-1 and Ls-2 were significantly more sensitive to trypsin treatment than the other two. The comparison of progeny virus titres after replication in alveolar macrophages allowed further differentiation among variants. Ls-1 and Ls-2 had similar titres in cultures infected with high virus input but in cultures infected with low virus input (0.1 TCID50/cell) Ls-1 produced higher titre. The difference in titres at 48 h post-infection was statistically significant (P less than 0.05). The cytopathogenicity for macrophages of the strains was correlated with their virulence for pigs, Stanley strain being the most cytopathogenic and NIA-4 the least. PMID- 2555109 TI - Further studies on passive immunization of newborn calves against rotaviral infection. AB - An inactivated rotavirus vaccine was prepared with an adjuvant which gives a water-in-oil emulsion. The vaccine when injected into cows in their last month of pregnancy, proved to be as effective as the traditional vaccine, prepared with the Freud's incomplete adjuvant, in enhancing the lactogenic immunity. Thus, feeding experimentally infected calves with colostrum and first milk from vaccinated cows prevented diarrhea and reduced significantly rotavirus shedding. Because of its low grade viscosity the water-in-oil emulsifier facilitates inoculation of the vaccine and is therefore recommended as an adjuvant in the preparation of inactivated rotavirus vaccine. PMID- 2555111 TI - Cell biology of antigen presentation by B cells. PMID- 2555110 TI - The emerging importance of IgE and Fc receptors for IgE (Fc epsilon R) in the regulation of B cell activity. PMID- 2555112 TI - Oxygen sensing and erythropoietin gene regulation. PMID- 2555113 TI - Characteristics of patients with primary lung cancer diagnosed at age of 50 years or younger. AB - We examined the extrinsic and intrinsic characteristics of patients with primary lung cancer diagnosed at early ages and compared them with those of older patients. Significant differences in gender distribution and histologic cell type were present. Cigarette smoking was an important etiologic factor in both groups. PMID- 2555114 TI - Brain metastases in patients with limited small cell lung cancer achieving complete remission. Correlation with TNM staging. AB - We reviewed the brain metastases, after treatment, of 45 patients with limited small cell lung cancer who achieved complete remission by radiochemotherapy or curative operation. No patient received prophylactic cranial irradiation. The incidence of subsequent brain metastases was classified according to pretreatment staging as follows: two of 13 (15 percent) patients in stage I; two of ten (20 percent) in stage II; nine of 17 (53 percent) in stage IIIa; and four of five (80 percent) in stage IIIb. The brain metastases occurred from seven to 29 months after the start of treatment, and the median time of the occurrence was 13 months. Of 17 patients who developed brain metastases and who subsequently received cranial irradiation, there were two in whom relapse had occurred at no other site except the brain and who survived 26 and 79 months after the relapse, respectively. These data indicate that not all patients with limited SCLC achieving CR due to treatment necessarily benefit from PCI. PMID- 2555115 TI - Copy number and distribution of P and I mobile elements in Drosophila melanogaster populations. AB - The distribution of the number of copies of P and I transposable elements per genome was investigated by in situ hybridization for a large set of Drosophila melanogaster strains. These included the P, Q and M' types of the P-M system of hybrid dysgenesis. P element copy number varied widely (range 5-59). P and Q strains had around 40 copies whereas M' strains generally had lower numbers (between 5 and 35) with one extreme value (52). The copy number of I elements appeared to be precisely regulated, as no strains were found outside the 15 +/- 5 range. The number of copies of the two families were independent. An excess of P copies on the X chromosome compared with the autosomes was found for the P and Q strains, but not for M' strains. Among X-inserted P sites, a very high frequency of occupation was found at the tip of the X chromosome (cytological site 1A), especially for P and Q strains. The possible regulatory role in the P-M system of X-inserted P sites is discussed. PMID- 2555117 TI - [The relationship between changing of anti-HAV in population and prevalence of hepatitis A]. AB - Hepatitis A was prevalent in 43 villages of the suburbs of Jinan, Shandong Province in 1986. The annual morbidity rate was 1,104.46/100,000. The authors studied the development of anti-HAV among children of under 14 years of age during the early epidemic and post-epidemic periods, compared the prevalence of anti-HAV between epidemic and non-epidemic villages, and analysed the types of HAV infection. The positive rate of anti-HAV was 35.42% during the early epidemic period and was 82.1% during the post-epidemic period, while it was 79.75% in non epidemic villages. The result indicates that, in rural area, hepatitis A is chiefly influenced by the prevalence of anti-HAV in the population. If it remains at about 80%, the occurrence of an epidemic can be prevented. The ratio of clinical, subclinical and apparent infection during the epidemic was 1:0.75:0.81. PMID- 2555118 TI - [A cohort study on the relation of primary liver cancer and HBsAg positivity in Guangxi Province]. PMID- 2555116 TI - The beta heterochromatic sequences flanking the I elements are themselves defective transposable elements. AB - Phylogenetic studies suggest that mobile element families are unstable components of the Drosophila genome. Two examples of immobilization of a transposable element family are presented here: as judged by their constant genomic organization among unrelated strains, the F and I element families have been respectively immobilized for a long time in D. simulans and in the reactive D. melanogaster strains (these are the laboratory strains which escaped the recent I invasion of D. melanogaster natural populations). All the elements of these defective families are located in the beta heterochromatic portion of the genome. Moreover, most if not all of the beta heterochromatic sequences into which the defective I elements are embedded are themselves non-mobile members of various nomadic families such as mdg 4, 297, 1731, F and Doc. These results are discussed with special emphasis on the possible nomadic origin of beta heterochromatin components and on the mechanisms of evolutionary turnover of the transposable element families. PMID- 2555120 TI - Role of rat large intestine in reducing diarrhea after 50% or 80% distal small bowel resection. AB - The effects of intestinal resection on several intestinal parameters have been studied in the large intestine of rats one month after the surgical operation. The results show that both 50% and 80% distal small bowel resection increased net fluid absorption and mucosal permeability and caused expansion of the intercellular spaces of the large intestine. The increase in net fluid absorption was dependent upon the extent of the intestine removed. The cAMP and cGMP content of cecal and colonic mucosa were significantly reduced after jejunoilectomy. Changes in nucleotide levels were dependent on the length of the intestine resected. On the other hand, mucosa Na,K-ATPase specific activity was only increased in the cecum after 80% intestinal resection. The results are discussed in terms of adaptation to prevent diarrhea. PMID- 2555119 TI - Endothelial cell growth regulation by PGE1 analog misoprostol and indomethacin. AB - The role of prostaglandins on growth regulation of different cell types was investigated. We studied the effect of exogenous misoprostol in the presence or absence of indomethacin on the cell growth kinetics. Our results clearly show that misoprostol strongly inhibited the growth of several cancer cells but only slightly affected that of endothelial cells. Moreover, indomethacin alone (which decreases PG synthesis) had little effect on DNA synthesis in endothelial cells or C6 cells but inhibited colonic cancer cells. Conversely, misoprostol stimulated DNA synthesis in endothelial or C6 cells preincubated with indomethacin. This phenomenon was not observed with colonic cancer cells. The role of NSAIDs in the field of cytoprotection is discussed. PMID- 2555121 TI - Changes in Na,K-ATPase, sodium ion, and glucose transport in isolated enterocytes in an experimental model of malabsorption. AB - Nippostrongylus brasiliensis infection of the rat resulted, at day 10 of infection, in decreased levels of jejunal enterocyte sodium-potassium-activated adenosine triphosphatase (Na,K-ATPase) and potassium-activated p-nitrophenyl phosphatase (K-pNPPase) activities. Parallel decreases occurred in active sodium efflux from jejunal enterocytes in the presence and absence of actively transported monosaccharides. Ileal enterocyte Na,K-ATPase and K-pNPPase activities were significantly increased, as was active sodium efflux. In contrast to controls, the presence of monosaccharides produced a stimulation of active sodium efflux from ileal enterocytes derived from infected rats. Enzyme and sodium transport changes in the jejunal enterocytes probably reflect cellular immaturity. Functional changes in ileal enterocytes probably represent a compensatory phenomenon. PMID- 2555122 TI - T-lymphocyte subsets in gut and blood of liver transplant recipients with and without cytomegalovirus gastroenteritis. AB - The effects of orthotopic liver transplantation (OLTx) and cytomegalovirus (CMV) gastroenteritis on the type of mononuclear cells within the upper gastrointestinal tract were determined. Nineteen liver transplant recipients were studied both before and after transplantation. Each underwent a pan-upper gastrointestinal endoscopy with biopsy of the antrum and duodenum before and four weeks following liver transplantation. A panel of monoclonal antibodies prepared against HLA-DR, NK, IL-2R, T11, T4, T8, and B1 cell surface antigens was used to examine the tissues. Before OLTx, none of the 19 subjects studied had clinical or histologic evidence for CMV gastroenteritis. Following OLTx, five of the 19 subjects had CMV gastroenteritis. The number of HLA-DR positive staining lymphocytes present in biopsies obtained post-OLTx was significantly greater (P less than 0.005) than those present in biopsies obtained pre-OLTx regardless of the presence or absence of CMV gastroenteritis. No difference in the intensity of HLA-DR antigen expression between pre- and post-OLTx biopsies and those with and without CMV gastroenteritis was evident. No difference in the number of natural killer (NK) cells and the number of cells expressing the interleukin-2 receptor (IL-2R) was evident between biopsies obtained pre- and post-OLTx. In contrast, the number of T lymphocytes bearing the T11, T4, and T8 markers and the calculated T4/T8 ratio differed between biopsies obtained pre- and post-OLTx and between those positive for CMV gastroenteritis post-OLTx and those without evidence for CMV gastroenteritis either before or after OLTx, although these changes were not consistent throughout the gastrointestinal tract.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555123 TI - Diagnostic accuracy of fine-needle aspiration biopsy in patients with hepatocellular carcinoma. AB - The present study was undertaken to investigate the diagnostic usefulness of fine needle aspiration biopsy (FNAB) in a large series of patients with hepatocellular carcinoma (HCC) seen over a 1-year period. During 1986, ultrasonographically guided percutaneous FNAB was performed in 72 patients with suspected HCC. A final diagnosis of HCC was made in 58 patients. The presence or absence of HCC was ascertained by histological examination and/or by other diagnostic procedures (alpha 1-fetoprotein, computed tomography, arteriography) and by clinical follow up (repeated ultrasonographic controls) and/or by surgery or necropsy. A total of 61 FNABs were carried out in these 58 patients. Only 42 (69%) of the 61 FNABs allowed the diagnosis of HCC. This moderate diagnostic sensitivity was not related to tumor size. Only one false positive result was observed in the non-HCC group. Therefore, the diagnostic specificity of FNAB for HCC was 93%, with a positive predictive value of 97% and a negative predictive value of 40%. These results show that FNAB is a useful diagnostic technique in patients with HCC. However, these data also show that there is a large proportion (31%) of subjects with false negative results. Therefore, we suggest that further efforts should be made to improve the diagnostic accuracy of this procedure. PMID- 2555124 TI - The different contributions of prostaglandins (E1, E2, F2 alpha, D2) to the tone and neurogenic response of bovine iris sphincter muscle. AB - The function and sites of action of prostaglandins (PGs) vary in different animal species and tissues. In this study the influence of PGs (E1, E2, F2 alpha, D2) on muscle tone and nerve-mediated contraction was investigated in isolated bovine iris sphincter muscles. None of these PGs exogenously applied influenced the neuromuscular transmission. By contrast, after treatment with indomethacin, all PGs tested contracted the muscle much more than in the absence of indomethacin and under these conditions the PGs potentiated responses to cholinergic nerve stimulation. Their ED50 were (2.2 +/- 0.2) x 10(-7) M for PGE1, (6.7 +/- 0.3) x 10(-8) M for PGE2, and (7.3 +/- 0.4) x 10(-8) M for PGF2 alpha. PGE1 acted both on nerves and the muscle cell. PGE2 had its influence mostly via nerves. Whereas PGF2 alpha was less potent in the absence of indomethacin, PGF2 alpha had much more potent action primarily on nerves and partly on muscles after treatment with indomethacin. High concentrations of PGD2 had both pre- and post-junctional action with accompanying weak contraction of the muscle. Thus the degrees of pre- and post-junctional involvement were different from one another. There is a possibility that the application of these PGs alone masked the role of such endogenous agents. In order to understand and clarify the site and action of PGs, pretreatment with indomethacin may be useful in the iris muscle. In conclusion, PGs modulate cholinergic activity in the bovine iris sphincter muscles, as well as regulate the muscle tone. PMID- 2555125 TI - [Visceral leishmaniasis (kala-azar). A rare differential diagnosis of splenomegaly and pancytopenia]. AB - A 53-year-old man developed a septic fever up to 40 degrees C, pancytopenia and hepatosplenomegaly after a holiday in Spain. Administration of piperacillin and amikacin was ineffective, but the fever subsided and partial haematological remission occurred when 1 mg/kg methylprednisolone daily was added. After six months his general condition worsened and pancytopenia with typical inclusion bodies in bone-marrow macrophages was noted, leading to the diagnosis of visceral leishmaniasis (Kala-Azar). The diagnosis was confirmed by serological tests. The causative organism was eliminated and the abnormal findings regressed during treatment with sodium stibogluconate, at first 600 mg/d for two weeks, then 850 mg/d over 16 days, interrupted for 14 days because of side effects. PMID- 2555126 TI - Computed tomography for determining pathways of extension and a staging and treatment system for juvenile angiofibromas. AB - Computed tomography (CT) has changed the preoperative assessment protocol of juvenile angiofibromas as it shows the extent of the tumor and its pathways of extension. Hence, operation can be done sooner and with minimal blood loss. Using CT scanning in 22 patients with nasopharyngeal angiofibromas, we found different pathways of extension of the tumor. We report a new staging system and surgical approaches, based on the stage of the tumor. PMID- 2555127 TI - In the mouse, the activation of the hypothalamic-pituitary-adrenal axis by a lipopolysaccharide (endotoxin) is mediated through interleukin-1. AB - The mechanisms through which endotoxin stimulates the hypothalamic-pituitary adrenal axis are not well understood. In the studies reported here we tested the hypothesis that endotoxin increases plasma ACTH levels by releasing interleukin-1 (Il-1). Two experimental tools reported to interfere with the biological activity of IL-1 were used: antibodies directed against IL-1 receptors and alpha MSH. In a first series of experiments, adult male mice were injected with a lipopolysaccharide (LPS; 25 micrograms), antibodies against IL-1 receptor, alpha MSH (1-30 micrograms), or LPS and either IL-1 antibodies or alpha MSH. All treatments were administered ip. The endotoxin LPS caused a marked increase in plasma ACTH levels, measured 6 h later. Both alpha MSH and the Il-1 receptor antibodies, while having no effect by themselves, significantly (P less than or equal to 0.01) blocked LPS-induced ACTH release. In a second series of experiments, mice were injected ip with 500 ng recombinant human (rh) Il-1 alpha or 100 ng rhIl-1 beta in the presence or absence of alpha MSH (1-30 micrograms, ip). While not altering ACTH secretion induced by rhIl-1 alpha, 10-30 micrograms alpha MSH significantly (P less than or equal to 0.01) interfered with the effect of rhIl-1 beta. These results suggest 1) that endotoxin activates the hypothalamic-pituitary-adrenal axis through a mechanism involving the activation of interleukin-1 receptors; and 2) that the effect of rhIl-1 beta, but not alpha, on ACTH secretion by the mouse can be partially blocked by alpha MSH. PMID- 2555128 TI - Glucocorticoid and 1,25-dihydroxyvitamin D modulate the degree of adenosine 3',5' monophosphate-dependent protein kinase isoenzyme I and II activation by parathyroid hormone in rat osteosarcoma cells. AB - Glucocorticoid increases and 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] decreases PTH activation of adenylate cyclase and cAMP-dependent protein kinase in rat osteosarcoma cells (ROS 17/2.8). Since selective cAMP-dependent protein kinase isoenzyme activation may account for specific physiological hormonal responses, we investigated steroid effects on activation of isoenzymes I and II in response to PTH using a new ion exchange separation procedure. Pretreatment of cells for 2 days with the glucocorticoid triamcinolone acetonide (TRM) or 1,25-(OH)2D3 altered the degree of cAMP-dependent protein kinase isoenzyme activation by PTH in accordance with their modulation of intracellular cAMP accumulation, but did not alter the amount of each isoenzyme present or the order in which isoenzymes I and II were activated. In all treatment groups isoenzyme I was preferentially activated by low doses of PTH, while high concentrations activated both isoenzymes, as predicted by the relative affinities of each isoenzyme for cAMP. Glucocorticoid reduced the concentration of bovine PTH-(1-34) required for maximal activation of isoenzyme I from 1 to 0.05 ng/ml and that required for activation of isoenzyme II from 10 to 1 ng/ml. This effect was abolished by simultaneous treatment of cells with 1,25-(OH)2D3. At doses of PTH that caused partial activation (0.05-0.1 ng/ml for isoenzyme I; 1 ng/ml for isoenzyme II), 1,25-(OH)2D3 treatment attenuated this activation. In all groups both isoenzymes were fully activated by 100 ng/ml PTH. Control experiments demonstrated that isoenzyme activation is not a result of cell disruption over the range of PTH doses that regulation by steroid hormone was observed. These results extend our studies on modulation of the cAMP pathway by steroid hormones and make it feasible to correlate selective isoenzyme activation with specific responses to PTH. PMID- 2555129 TI - Characterization of a temperature-sensitive beta-endorphin-secreting transformed endometrial cell line. AB - An endometrial cell line (HRE-H9) was established and characterized to study the mechanism by which gene expression of POMC-derived peptides is controlled in uterine tissues. The HRE-H9 cell line was isolated by transforming primary rabbit endometrial cell cultures, derived from hCG-treated pseudopregnant rabbits, with a temperature-sensitive A209 mutant (tsA209) simian virus 40 at a permissive temperature (33 C). The resulting cells exhibited temperature sensitivity in growth and synthesis of immunoreactive beta-endorphin (ir-beta END). The ir-beta END present in the cell extracts and culture media was assayed by a specific beta END RIA. Sephadex G-50 gel filtration chromatography of the transformed cell extracts showed three peaks of beta END immunoreactivity. The first peak eluted at the void volume, the second peak coeluted with the beta-lipotropin standard, and the third peak coincided with the porcine beta END standard, ir-beta-END was also detectable in endometrial culture media, suggesting that the transformed endometrial cells secreted POMC-derived peptides. Our data indicate that the tsA209 mutant virus-transformed endometrial cell line provides a suitable model for study of the synthesis and regulation of POMC-derived peptides in extrapituitary tissues. PMID- 2555130 TI - Elevation of hypophysial portal concentrations of adrenocorticotropin secretagogues after fornix transection. AB - Glucocorticoid feedback inhibition at the level of the brain is extremely complex, involving feedback at both hypothalamic and suprahypothalamic levels. The hippocampus has been implicated as a suprahypothalamic mediator of such feedback, based on numerous lesion, stimulation, and steroid implantation studies. These reports, however, predated the isolation and characterization of CRF and recognition of the multifactorial control of ACTH release. Thus, it is not clear which hypothalamic ACTH secretagogues are under inhibitory control of the hippocampus. To answer this, we measured hypophysialportal concentrations of CRF, arginine vasopressin, and oxytocin in rats with fornix transections, which disrupt hippocampal communication with the hypothalamus. Hypophysial-portal blood was collected in rats exposed to either low or high circulating corticosterone concentrations in the presence or absence of the coincident stressor of hypotension. We observed that fornix transection produced hypersecretion of all three secretagogues. However, the pattern of hypersecretion differed for each as follows: 1) fornix transection did not affect either initial CRF secretion or the magnitude of the stress response, but made rats resistant to a high feedback signal during stress; 2) fornix transection led to initial arginine vasopressin hypersecretion, which remained sensitive to a high feedback signal; and 3) fornix transection led to initial oxytocin hypersecretion as well as resistance to a high corticosterone feedback signal during hypotension. PMID- 2555131 TI - Rate-sensitive glucocorticoid feedback inhibition of adrenocorticotropin and beta endorphin/beta-lipotropin secretion in rats. AB - In the present study, we present physiological evidence for rate-sensitive, fast feedback inhibition of secretion of ACTH and beta-endorphin (beta END)-related peptides. We used a 2 min restraint stress to physiologically increase plasma corticosterone, then examined the plasma responses of immunoreactive ACTH and beta END plus beta-lipotropin (beta END/beta LPH) to a subsequent restraint stress. After onset of this stress, plasma corticosterone increased from 2.5-10 min at a rate of 120 nM min-1, then remained at a peak from 10-15 min. A single 2 min restraint stress produced peak plasma levels of ACTH and beta END/beta LPH 2.5 min after onset of the stress, and these plasma concentrations declined after this initial stress at rates of 2.7 and 7.4 pM min-1, respectively. Application of a second restraint stress at the time of the peak corticosterone response produced plasma ACTH and beta END/beta LPH responses similar to those after the first stress. Application of a second stress during the period of significant rate-rise of corticosterone in plasma did not result in decreased incremental responses of plasma ACTH or beta END/beta LPH. However, the rates of decline of plasma ACTH and beta END/beta LPH of 7.6 and 32 pM min-1, respectively, from peak levels, were significantly greater after this second stress applied during the period of significant increase in plasma corticosterone concentration than the corresponding rates of decline observed after the initial stress or after a subsequent stress applied at the peak of plasma corticosterone. These differences in rates of decline of plasma ACTH or beta END/beta LPH appear to reflect differences in secretion rate rather than clearance, since disappearance of [125I]ACTH1-24 was not different after an initial vs. subsequent stress. In contrast to these data from intact rats, initial and subsequent stresses did not show different rates of decline of plasma ACTH or beta END/beta LPH in adrenalectomized rats. In conclusion, the stress-induced rate rise of glucocorticoid provides a negative feedback signal which serves to terminate and limit the duration, but not the peak, of the responses of POMC-derived peptides to subsequent stress. PMID- 2555132 TI - Modulation of basal and corticotropin-releasing factor-stimulated proopiomelanocortin gene expression by vasopressin in rat anterior pituitary. AB - In several anterior pituitary hormone systems, factors that stimulate hormone release also stimulate hormone biosynthesis. In the corticotropes of the anterior pituitary, CRF stimulates ACTH release as well as cAMP accumulation and transcription of the POMC gene. Arginine vasopressin (AVP) is a well documented coregulator of ACTH release and potentiates CRF-stimulated cAMP accumulation. The present studies were undertaken to determine if AVP also potentiates the early effects of CRF on POMC gene expression in rat anterior pituitary primary cultures. We have measured the levels of the POMC primary transcript, processing intermediate, and mature mRNA in nuclear RNA samples and POMC mRNA in cytoplasmic RNA samples as well as ACTH release after treatment with CRF and/or AVP. After a 30-min treatment with 0.5 nM CRF, POMC primary transcript levels were increased by 200-400%. Thirty-minute or 1-h treatments with AVP alone (10 or 100 nM) did not affect primary transcript levels, but increased the amount of the processing intermediate, while a 2-h incubation with 100 nM AVP significantly decreased POMC primary transcript levels. The effects of CRF on the POMC primary transcript and nuclear processing intermediate were not potentiated by cotreatment with AVP, although a potentiation of CRF-stimulated ACTH release was observed. POMC nuclear and cytoplasmic mRNA levels were not affected by these 2-h or less treatments, while an 18-h incubation with 0.5 nM CRF alone or in combination with 100 nM AVP increased POMC cytoplasmic mRNA levels to about 140% of vehicle-treated control values. When a 1-h AVP treatment (100 nM) preceded presentation of CRF, we observed an attenuation of the stimulation of POMC primary transcript and processing intermediate levels by the 1-h CRF treatment. The effects of CRF on POMC primary transcript and processing intermediate levels in these experiments are consistent with a CRF-induced increase in POMC gene transcription. Our findings that AVP decreased primary transcript levels and attenuated the effects of CRF on this end point suggest that: 1) AVP clearly does not act in a synergistic manner with CRF to stimulate POMC gene expression; and 2) AVP possibly affects RNA stability and/or processing. PMID- 2555133 TI - Receptor-mediated endocytosis and differential synthesis of mannose 6-phosphate receptors in isolated spermatogenic and sertoli cells. AB - Mannose 6-phosphate (Man 6-P) receptors participate in the targeting of both newly synthesized and extracellular acid hydrolases to lysosomes, and also may mediate the effects of a number of growth factors. In this study, Man 6-P receptors were isolated from [35S]methionine-labeled germ cells and Sertoli cells by phosphomannan-Sepharose affinity chromatography and were analyzed by polyacrylamide gel electrophoresis and autoradiography. Mouse pachytene spermatocytes and round spermatids isolated by unit gravity sedimentation synthesized predominantly the 46,000 mol wt (Mr) cation-dependent Man 6-P receptor and only low levels of the 270,000 Mr cation-independent Man 6-P receptor. In contrast, Sertoli cells synthesized substantial amounts of the cation-independent Man 6-P receptor, but little of the cation-dependent receptor. To determine if these receptors function on the cell surface, we have monitored Man 6-P receptor-mediated endocytosis in isolated germ cells and Sertoli cells. In pachytene spermatocytes and round spermatids, [125I]Man 6-P-bearing ligands were internalized and processed to lower Mr forms by an endocytic mechanism that was time dependent, proportional to cell number, and inhibited by Man 6-P. Like germ cells, Sertoli cells in primary culture endocytosed radiolabeled Man 6-P bearing ligands at levels that were about 10% of the endocytic activity measured for 3T3 fibroblasts. This low endocytic activity correlates with the levels of the cation-independent Man 6-P receptor synthesized by germ cells, but not with the higher levels synthesized by Sertoli cells. Membrane binding assays verified the high steady state levels of the cation-independent Man 6-P receptor in Sertoli cells, suggesting that the low endocytic activity detected in these cells may result from restricted expression of the cation-independent receptor on the cell surface. These results indicate that both spermatogenic and Sertoli cells have surface Man P-6 receptors capable of mediating endocytosis. However, these cells exhibit marked differences in the expression of the cation-independent and dependent Man 6-P receptors, perhaps reflecting differential roles of these receptors in protein trafficking and/or intercellular communication during germ cell differentiation. PMID- 2555134 TI - Estrogen regulation of an eosinophil chemotactic factor in the immature rat uterus. AB - Associated with the generalized uterine growth stimulated by estradiol in the rat are specific responses including messenger RNA (mRNA) synthesis, protein synthesis, and peroxidase activity. The increase in peroxidase activity, although sensitive to inhibitors of RNA and protein synthesis, results from an estradiol stimulated influx of eosinophils into the uterus. We postulated the existence of an estradiol-regulated uterine chemotactic factor, testing this by an in vitro chemotactic assay with eosinophils isolated from mature rats. Treatment of immature rats with 1 microgram estradiol for 24 h resulted in a significant increase in eosinophil chemotaxis compared to uterine extracts of vehicle-treated rats. This increase was seen as early as 3 h after estradiol administration and was maximal at 24 h. The magnitude of the chemotactic response was dependent on the dose of estradiol administered (1-100 micrograms). Estrone or estriol treatment resulted in chemotactic activity greater than control but less than estradiol. Direct addition of estradiol to extracts of control animals did not increase chemotaxis. The estradiol-stimulated chemotaxis was blocked by in vivo treatment with the antiestrogen tamoxifen and by inhibitors of RNA and protein synthesis. Analysis of extracts from estradiol-treated uteri shows that the chemotactic factor is heat labile, pronase sensitive, and has a mass of approximately 20 kilodaltons (kDa). These data suggest that the estradiol stimulated influx of eosinophils into the rat uterus is mediated by the synthesis, modification, or release of a protein whose synthesis is estradiol receptor mediated. PMID- 2555135 TI - Follicle-stimulating hormone receptor-mediated uptake of 45Ca2+ by proteoliposomes and cultured rat sertoli cells: evidence for involvement of voltage-activated and voltage-independent calcium channels. AB - We have previously reported incorporation into liposomes of Triton X-100 solubilized FSH receptor-G-protein complexes derived from purified bovine calf testis membranes. In the present study we have used this model system to show that FSH induces flux of 45Ca2+ into such proteoliposomes in a hormone-specific concentration-dependent manner. FSH, inactivated by boiling, had no stimulatory effect on 45Ca2+ flux, nor did isolated alpha- or beta-subunits of FSH. Addition of GTP (or its analogs 5'-guanylylimidodiphosphate and guanosine-5'-O-[3 thiotriphosphate]) or sodium fluoride (in the presence or absence of GTP or its analogs) failed to induce 45Ca2+ flux into proteoliposomes, suggesting that the uptake of 45Ca2+ was receptor, and not G-protein, related. Voltage-independent (ruthenium red and gadolinium chloride) and voltage-activated (methyoxyverapamil and nifedipine) calcium channel-blocking agents reduced FSH-stimulated 45Ca2+ flux into proteoliposomes to control levels. FSH also induced uptake of 45Ca2+ by cultured rat Sertoli cells. Ruthenium red and gadolinium chloride had no effect on basal levels of 45Ca2+ uptake or estradiol secretion by cultured rat Sertoli cells, nor did methoxyverapamil or nifedipine. All four calcium channel blockers, however, were able to reduce FSH-induced 45Ca2+ uptake to basal levels and FSH stimulated conversion of androstenedione to estradiol by up to 50%, indicating an involvement of Ca2+ in FSH-stimulated steroidogenesis. Our results suggest that the well documented changes in intracellular calcium levels consequent to FSH binding may be due, at least in part, to an influx of calcium through FSH receptor-regulated calcium channels. PMID- 2555136 TI - Expression of hepatic transforming growth factor receptors during late gestation in the fetal rat. AB - Transforming growth factor-alpha (TGF alpha) promotes DNA synthesis in adult rat hepatocytes, an effect opposed by picomolar concentrations of TGF beta. Recently, the presence of these growth factors in fetal rat liver has been demonstrated. Since a regulatory role for TGF alpha and TGF beta in fetal hepatic growth requires the presence of high affinity receptors, the receptors for these hormones were studied in membranes from normal fetuses at 17-21 days gestational age and growth-retarded fetuses of mothers fasted for 48 h. Fetal liver membranes bound [125I]epidermal growth factor ([125I]EGF) with high affinity (Kd = 1-2 nM). TGF alpha could compete with EGF for the same binding site, albeit at 4-fold lower affinity. EGF receptor number increased from nearly undetectable levels at 17 days to adult levels (0.15-0.3 nmol/mg membrane protein) by 21 days. Affinity labeling of fetal liver membranes with [125I]TGF alpha identified the 170,000 mol wt (Mr) EGF receptor. The intensity of labeling correlated with EGF receptor number based on binding analyses. TGF beta bound to fetal liver membranes with high affinity (Kd = 30 pM) and at a level (20-30 pmol/mg throughout late gestation) that was 3-fold higher than in adult liver. Affinity labeling of fetal hepatic membranes with [125I]TGF demonstrated high affinity 85,000 Mr TGF beta receptors and lower affinity 66,000 and 130,000 Mr receptors. Although TGF beta binding did not change with advancing gestation, affinity labeling of the 85,000 Mr protein doubled from day 18 to day 21 and was decreased by 50% in fetuses from fasted mothers. These data, demonstrating the presence and regulation of the receptors for TGF alpha and TGF beta, support roles for these hormones in the regulation of fetal hepatic growth. PMID- 2555137 TI - Solubilization of high affinity corticotropin-releasing factor receptors from rat brain: characterization of an active digitonin-solubilized receptor complex. AB - The binding characteristics of CRF receptors in rat frontal cerebral cortex membranes solubilized in 1% digitonin were determined. The binding of [125I]Tyro ovine CRF ([125I]oCRF) to solubilized membrane proteins was dependent on incubation time, temperature, and protein concentration, was saturable and of high affinity, and was absent in boiled tissue. The solubilized receptors retained their high affinity for [125I] oCRF in the solubilized state, exhibiting a dissociation constant (KD) of approximately 200 pM, as determined by direct binding saturation isotherms. Solubilized CRF receptors maintained the rank order of potencies for various related and unrelated CRF peptides characteristic of the membrane CRF receptor: rat/human CRF congruent to ovine CRF congruent to Nle21,38 rat CRF greater than alpha-helical oCRF-(9-41) greater than oCRF-(7-41) much greater than vasoactive intestinal peptide, arginine vasopressin, or the substance-P antagonist. Furthermore, the absolute potencies (Ki values) for the various CRF-related peptides in solubilized receptors were almost identical to those observed in the membrane preparations, indicating that the CRF receptor retained its high affinity binding capacity in the digitonin-solubilized state. Chemical affinity cross-linking of digitonin-solubilized rat cortical membrane proteins revealed a specifically labeled protein with an apparent mol wt of 58,000 which was similar to the labeled protein in native membrane homogenates. Although solubilized CRF receptors retained their high affinity for agonists, their sensitivity for guanine nucleotide was lost. Size exclusion chromatography substantiated these results, demonstrating that in the presence or absence of guanine nucleotides, [125I]oCRF labeled the same size receptor complex. These data suggest that either the guanine nucleotide-binding protein (Ns) is tightly associated with the CRF receptor after solubilization and is insensitive to guanine nucleotides, or that high affinity binding for soluble CRF receptors is not dependent on the coupling of a guanine nucleotide-binding protein. The solubilization of CRF receptors from membranes in digitonin should allow for the more complete molecular and functional characterization of CRF-mediated events and purification of the receptor. PMID- 2555138 TI - Tumor necrosis factor and interleukin-1 are potent inhibitors of angiotensin-II induced aldosterone synthesis. AB - Cytokines such as tumor necrosis factor (TNF) and interleukin-1 (IL-1), mediate many inflammatory and cellular responses. However, the effects of TNF and IL-1 on basal and angiotensin-II (AII)-stimulated aldosterone synthesis are not known. We studied the effect of recombinant and purified TNF and IL-1 on basal as well as AII-, ACTH-, and K+-induced aldosterone synthesis in isolated rat adrenal glomerulosa cells. Since we have previously shown that AII action is mediated by activation of the 12-lipoxygenase (12LO) pathway of arachidonic acid, we also evaluated the effects of these cytokines on the 12LO product 12 hydroxyeicosatetraenoic acid (12HETE) using a validated RIA technique. TNF at 2.5 and 5.0 ng/ml produced a dose-dependent inhibition of AII-induced aldosterone synthesis [AII, 39.0 +/- 3.3 ng/10(6) cells.h; AII plus TNF (5.0 ng/ml), 14.3 +/- 1.6; P less than 0.001 vs. AII; AII plus TNF (2.5 ng/ml), 24.7 +/- 3.2; P less than 0.01 vs. AII]. Similarly, TNF at 5.0 ng/ml also attenuated the stimulatory effect of ACTH (10(-9) M). However, K+-induced aldosterone synthesis was not altered. TNF also did not alter basal aldosterone levels. AII, as previously shown, stimulates 12HETE synthesis (basal, 608 +/- 114 pg/10(5) cells.h; versus AII, 1268 +/- 197; P less than 0.02). TNF at concentrations of 1.0-5.0 ng/ml produced a dose-dependent inhibition of AII stimulatory action on 12HETE synthesis [AII plus TNF (1.0 ng/ml), 650 +/- 26 pg, P less than 0.03 vs. AII; AII plus TNF (5.0 ng/ml), 390 +/- 46; P less than 0.01 vs. AII plus TNF (1.0 ng/ml)]. In addition, 12HETE at 10(-8) M completely restored the effects of AII during blockage by TNF. Purified human IL-1 (75% beta, 25% alpha) as well as recombinant human IL-1 beta at concentrations as low as 50 pg/ml inhibited AII-induced aldosterone synthesis. IL-1 beta did not alter ACTH- or K+-induced aldosterone synthesis and, in fact, had a tendency to potentiate ACTH effects. These results suggest that the cytokines TNF and IL-1 are potent inhibitors, particularly of AII action in the adrenal glomerulosa cell. Therefore, local or systemically produced TNF or IL-1 may be important negative modulators of aldosterone synthesis. PMID- 2555140 TI - Preservation of sodium-dependent iodide transport activity by methimazole and mercaptoethanol in phospholipid vesicles containing thyroid plasma membranes: with evidence of difference in the action of perchlorate and thiocyanate. AB - The effect of methimazole (MMI) and 2-mercaptoethanol (ME) on I-transport was studied using phospholipid vesicles (P-vesicles) made from porcine thyroid plasma membranes and soybean phospholipids by sonication. 1. When buffer solutions contained either 1 mM MMI or 2 mM ME, I-uptake by P-vesicles in the presence of external Na+ was apparently higher than that in the absence of external Na+. Na+ dependent I- uptake was inhibited by both C1O4- and SCN- added externally. 2. When PM was treated with 4 mM N-ethylmaleimide prior to preparation of P vesicles, the activity of Na+-dependent I- transport was completely lost even when P-vesicles were incubated in the presence of ME. 3. When neither MMI nor ME was added to buffers, I- uptake in the presence of external Na+ was not at all higher than that in the absence of external Na+. In these instances, however, I- uptake was much higher compared than the baseline uptake in the presence of MMI or ME, and was inhibited by external SCN- and not by C1O4- without relation to external Na+. These data indicate that MMI or ME has two distinct effects on our model system of I- transport. The one is preservation of the Na+-dependent I- transport activity by protecting a sulfhydryl group, and the other is reduction of nonspecific I- binding to P-vesicles. In addition, C1O4- is a more specific inhibitor of thyroid I- transport than SCN-, when non-specific I- oxidation is imperfectly prevented. PMID- 2555139 TI - Selective inhibition of angiotensin-II-mediated aldosterone secretion by 5 hydroxyeicosatetraenoic acid. AB - We have previously demonstrated that the 12-lipoxygenase (12-LO) pathway plays a key role in angiotensin-II (AII)-dependent aldosterone production. In the present study we examined the role of the 5LO pathway on AII-induced aldosterone secretion in rat glomerulosa cells in vitro. The 5LO product 5 hydroxyeicosatetraenoic acid (5HETE) and its unstable precursor 5 hydroxyperoxyeicosatetraenoic acid did not significantly alter basal aldosterone secretion in concentrations from 10(-9)-10(-7) M. In contrast, 5HETE reduced peak AII-induced aldosterone production from 59.1 +/- 9.0 to 37.96 +/- 7.2 ng/10(6) cells (P less than 0.01). This was accompanied by inhibition of the AII stimulated rise in 12HETE production (10(-9)M AII, 160 +/- 4% of control; 10(-9) M AII plus 10(-7) M 5HETE, 90 +/- 1% of control production). However, 5HETE had no effect on the aldosterone response to potassium or ACTH, secretagogues that cause no activation of the 12LO pathway. These results suggest that the 5LO product 5HETE can selectively modulate AII-dependent aldosterone secretion. Further, the selective inhibitory effect of 5HETE on the AII effect in rat glomerulosa cells may be exerted by blockade of arachidonate metabolism via the 12LO pathway. These results suggest that the 5LO pathway may negatively modulate AII action in the adrenal zona glomerulosa. PMID- 2555141 TI - Malignant lymphoma of the thyroid and Epstein-Barr virus. AB - We have investigated the specific immune response to Epstein-Barr virus (EBV) of peripheral blood mononuclear cells (PBMC) from patients with malignant lymphoma of the thyroid. Coculture of PBMC and EBV resulted in EBV cell transformation and regression which was assayed by an EBV-induced B cell focus-regression assay technique. The EBV had been isolated from mouthwash samples. The specific immune response to EBV by outgrowth inhibition in PBMC from untreated EBV-seropositive patients with malignant lymphoma was significantly decreased when compared to PBMC from EBV-seropositive healthy subjects (p less than 0.05). This observation is at least consistent with the possibility that B-cell proliferation after continuous or recurrent EBV infection could be a causative factor or may potentiate malignant lymphoma of the thyroid. PMID- 2555143 TI - Effects of exercise training on brown adipose tissue thermogenesis in ovariectomized obese rats. AB - The effect of exercise training on brown adipose tissue (BAT) thermogenesis was studied by measuring cytochrome oxidase activity, as a marker of mitochondrial abundance, mitochondrial guanosine-5'-diphosphate (GDP) binding, as an indicator of thermogenic activity and oxygen consumption in BAT in ovariectomized (OVX) obese rats and sham-operated rats. Six-week exercise training significantly suppressed body weight gain in OVX rats to the level of sedentary control rats, although food intake in exercise trained OVX rats increased more than in the sedentary OVX rats. Exercise training increased cytochrome oxidase activity, mitochondrial GDP binding and oxygen consumption in BAT in OVX rats, which were reduced in a sedentary condition, as well as in the control rats. These results suggest that exercise training potentiates BAT thermogenesis, which may contribute to the reduction of body weight in OVX obese rats. PMID- 2555142 TI - LHRH increases plasma 7B2 concentration in normal human subjects. AB - We studied the response of plasma 7B2 to LHRH and ovine corticotropin releasing hormone (o-CRH) in healthy young subjects. The plasma 7B2 concentration significantly increased from 78.3 +/- 7.5 (mean +/- SEM) to 102.0 +/- 6.0 ng/L (142.7 +/- 12.7% of the basal value; P less than 0.01) following iv administration of LHRH in seven young subjects. On the other hand, no increase in plasma 7B2 was found after iv administration of o-CRH in six young subjects. These results, together with our previous report of no increase in plasma 7B2 after administration of TRH and GHRH in young subjects, suggest that pituitary 7B2 may be present in gonadotrophs and be released only by LHRH in physiological conditions. PMID- 2555144 TI - Effects of chronic infusion of dibutyryl c-AMP or adenosine on luteal function in sheep. AB - Adenosine or vehicle; dibutyryl c-AMP, a c-AMP analogue, or vehicle in two separate experiments were infused through an indwelling cannula every four hours around the ovarian vascular pedicle of ewes unilaterally ovariectomized on day 8 postestrus. Adenosine or vehicle was infused from day 8 through 22 postestrus and dibutyryl-cAMP was infused from day 8 through 20 postestrus or until the ewes returned to estrus. Interestrous intervals were greater (p less than or equal to 0.05) in ewes receiving adenosine (27.3 +/- 2.4 days) than in control ewes (17.2 +/- 1.3 days). The length of the estrous cycle of ewes receiving dibutyryl c-AMP was greater (22.4 +/- 1.1; p less than or equal to 0.05) than in control ewes which averaged 16.7 +/- 0.6 days. Profiles of progesterone were different (p less than or equal to 0.05) for ewes receiving adenosine or dibutyryl c-AMP when compared to their respective controls. In addition, the overall mean concentrations of progesterone were greater (p less than or equal to 0.05) in dibutyryl c-AMP or adenosine-treated ewes than in controls. In a third experiment, infusions of adenosine or dibutyryl c-AMP intrauterine every 4 hours through a cannula from day 8 through 22 postestrus had no effect (p less than or equal to 0.05) on the interestrous interval or profiles of progesterone. It is concluded that dibutyryl c-AMP or adenosine in vivo can delay luteolysis and adenosine and c-AMP may play roles in luteal secretion of progesterone in sheep but are probably not the uterine embryonic antiluteolysin of early pregnancy in sheep. PMID- 2555145 TI - Isolated ACTH deficiency associated with transient thyrotoxicosis and hyperprolactinemia. AB - A 43-year-old woman with isolated ACTH deficiency in association with transient thyrotoxicosis is reported. The initial evaluation revealed that plasma ACTH and cortisol did not respond to corticotropin-releasing hormone (CRH) in the presence of hyperthyroxinemia and hyperprolactinemia. During the replacement therapy with dexamethasone, she developed transient hypothyroxinemia with persistent hyperprolactinemia. Although thyroid open biopsy did not show any evidence of autoimmune thyroiditis or subacute thyroiditis, the data appear to provide other evidence of a possible relationship between acute adrenal insufficiency and transient thyroid dysfunction. PMID- 2555146 TI - Pseudohypoparathyroidism showing positive phosphaturic and negative cyclic AMP excretion response to parathyroid hormone. AB - We report a patient with pseudohypoparathyroidism (PHP) in whom parathyroid hormone (PTH) infusion failed to produce an increase in urinary adenosine 3', 5' monophosphate (cAMP) excretion in spite of the positive urinary phosphate excretion. The dbcAMP infusion test showed almost the same increase in phosphate as in the E-H test, although high urinary cAMP excretion was detected. Furthermore, a PTH infusion test in combination with calcium antagonist (diltiazem) administration markedly increased phosphate excretion, whereas the response of urinary cAMP excretion also remained negative. After treatment with 1 alpha(OH)D3, phosphaturic response increased by at least 14.3 mg/2 h compared with that in the pretreatment period. Therefore, intra and extra cellular calcium seem to affect the phosphaturic response induced by PTH. PMID- 2555147 TI - The role of insulin in norepinephrine turnover and thermogenesis in brown adipose tissue after acute cold-exposure. AB - The role of insulin in norepinephrine turnover (NE) and thermogenesis in brown adipose tissue (BAT) after acute cold-exposure was studied using streptozocin (STZ)-induced diabetic rats. NE turnover was estimated by the NE synthesis inhibition technique with alpha-methyl-p-tyrosine. BAT thermogenesis was estimated by measuring mitochondrial guanosine-5'-diphosphate (GDP), cytochrome oxidase activity and mitochondrial oxygen consumption in BAT at an ambient temperature of 22 degrees C and during a six-hour cold-exposure at 4 degrees C. In insulin-deficient diabetic rats, the NE turnover, mitochondrial GDP binding, cytochrome oxidase activity and mitochondrial oxygen consumption in BAT at 22 degrees C were significantly reduced, compared with those of control rats. Treatment of STZ-induced diabetic rats with insulin prevented a decrease in NE turnover and BAT thermogenesis. Acute cold-exposure increased the NE turnover of BAT in insulin-deficient diabetic rats. The BAT thermogenic response to acute cold-exposure, however, did not occur in insulin-deficient diabetic rats. These results suggest that insulin is not essential in potentiating NE turnover in BAT after acute cold-exposure, but is required for cold-induced thermogenesis. PMID- 2555148 TI - Possible hyperaldosteronism and discrepancy in enzyme activity deficiency in adrenal and gonadal glands in Japanese patients with 17 alpha-hydroxylase deficiency. AB - We reviewed the pathophysiology of our previously reported female patient who had glucocorticoid-responsive hyperaldosteronism and was treated successfully with daily dose of dexamethasone (Dex) for 21 years. In this present study, the possibility that the patient may have 17 alpha-hydroxylase deficiency (17-OH-D) mainly in the adrenal could not be ruled out. We therefore reviewed 31 Japanese patients diagnosed as having 17-OH-D with suppressed plasma renin activity reported in Japan. Among these patients, 9 were found to have a high plasma aldosterone (Ald) concentration (PAC) (group I). Twenty-one patients had either normal or low-normal PAC and the remaining patient had low urine Ald (group II). The slight cross-reactivity of the anti-Ald-antibodies used with 17-deoxy steroids such as progesterone, 11-deoxycorticosterone and corticosterone which were increased in both groups did not explain the increased PAC in group I. In the patients in group I and group II with high-normal basal PAC, PAC further increased after ACTH and was suppressed by Dex. PAC in 2 group I patients, however, did not respond to angiotensin-II or angiotensin-III infusion. PAC in patients in group II with low or low-normal basal PAC responded equivocally to ACTH and Dex. The basal plasma cortisol in group I was lower than in group II, and plasma cortisol level after ACTH in group I appeared to remain at a lower level than that in group II patients. Among the study subjects, 28 showed a negative correlation between basal PAC and plasma cortisol. A possible discrepancy in the deficiency of 17 alpha-hydroxylase activity in adrenal and gonadal glands was also suggested in three 17-OH-D patients. The pathophysiology of Ald secretion and discrepancy in the deficiency of the enzyme activities in both glands in 17-OH-D patients was discussed. PMID- 2555149 TI - Chronic stress, leukocyte subpopulations, and humoral response to latent viruses. AB - Psychological stress has been shown to affect immune system status and function, but most studies of this relationship have focused on acute stress and/or laboratory situations. The present study compared total numbers of leukocytes and lymphocyte subpopulations (determined by flow cytometry) and antibody titers to latent and nonlatent viruses among a group of chronically stressed individuals living near the damaged Three Mile Island (TMI) nuclear power plant with those of a demographically comparable control group. Urinary catecholamine and cortisol levels were also examined. Residents of the TMI area exhibited greater numbers of neutrophils, which were positively correlated with epinephrine levels. The TMI group also exhibited fewer B lymphocytes, T-suppressor/cytotoxic lymphocytes, and natural killer cells. Antibody titers to herpes simplex were significantly different across groups as well, whereas titers to nonlatent rubella virus as well as IgG and IgM levels were comparable. PMID- 2555151 TI - A transforming function of the BARF1 gene encoded by Epstein-Barr virus. AB - We report a new rodent cell-transforming gene, presumably involved in viral replication, encoded by Epstein-Barr virus. We previously showed that the corresponding open reading frame BARF1 is transcribed before the onset of viral DNA synthesis, and translated into a 33 kd early polypeptide (p33). Here we show that recombinant plasmids containing the BARF1 induce morphological change, anchorage-independent growth and tumorigenic transformation of established mouse fibroblast lines. The BARF1-transformed cells and the tumour tissues isolated from new-born rats after injection of such transformed cell both express p33. Transforming activity was obtained from either the genomic fragment or the cDNA sequence. PMID- 2555150 TI - Neuraxin, a novel putative structural protein of the rat central nervous system that is immunologically related to microtubule-associated protein 5. AB - During screening of a rat spinal cord lambda gt11 cDNA library with poly- and monoclonal antibodies against the postsynaptic glycine receptor a cDNA was isolated which covers an open reading frame encoding a protein of calculated mol. wt 94 kd. Sequence analysis identified a novel type of neuron-specific protein (named neuraxin) which is characterized by an unusual amino acid composition, 12 central heptadecarepeats and putative protein and/or membrane interaction sites. The gene encoding neuraxin appears to be unique in the haploid rat genome and conserved in higher vertebrates. Northern blot and in situ hybridization revealed neuraxin mRNA to be expressed throughout the rodent central nervous system (CNS). In spinal cord, neuraxin transcripts were abundant in motoneurons which also expressed glycine receptor subunit mRNA. A bacterial fusion protein containing approximately 90% of the neuraxin sequence was found to specifically bind tubulin. Polyclonal neuraxin antibodies cross-reacted with microtubule-associated protein 5 (MAP5), and a monoclonal antibody against MAP5 recognized the neuraxin fusion construct. Based on these data we suggest that neuraxin is related to MAP5 and may be implicated in neuronal membrane-microtubule interactions. PMID- 2555152 TI - Identification of a common signal associated with cellular proliferation stimulated by four haemopoietic growth factors in a highly enriched population of granulocyte/macrophage colony-forming cells. AB - We have prepared a population of bone marrow cells that is highly enriched in neutrophil/macrophage progenitor cells (GM-CFC). Four distinct haemopoietic growth factors can stimulate the formation of mature cells from this population, although the proportions of neutrophils and/or macrophages produced varied depending on the growth factor employed: interleukin 3 (IL-3) and granulocyte/macrophage colony-stimulating factor (GM-CSF) stimulated the formation of colonies containing both neutrophils and macrophages; macrophage colony-stimulating factor (M-CSF) produced predominantly macrophage colonies; and granulocyte colony-stimulating factor (G-CSF) promoted neutrophil colony formation. Combinations of these four growth factors did not lead to any additive or synergistic effect on the number of colonies produced in clonal soft agar assays, indicating the presence of a common set of cells responsive to all four haemopoietic growth factors. These enriched progenitor cells therefore represent an ideal population to study myeloid growth-factor-stimulated survival, proliferation and development. Using this population we have examined the molecular signalling mechanisms associated with progenitor cell proliferation. We have shown that modulation of cyclic AMP levels has no apparent role in GM-CFC proliferation, whereas phorbol esters and/or Ca2+ ionophore can stimulate DNA synthesis, indicating a possible role for protein kinase C activation and increased cytosolic Ca2+ levels in the proliferation of these cells. The lack of ability of all four myeloid growth factors to mobilize intracellular Ca2+ infers that these effects are not achieved via inositol lipid hydrolysis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555153 TI - Three hormone receptor-like Drosophila genes encode an identical DNA-binding finger. AB - The putative finger domain of knirps (kni), a member of the gap class of segmentation genes, was used to isolate two sequence-related genes of Drosophila melanogaster under reduced stringency hybridization conditions. The two kni homologous genes map close to kni in the proximal portion of the third chromosome. One of them is the previously identified gene knirps-related (knrl), kni and knrl are spatially co-regulated in both early and late stages of embryogenesis. Their posterior domains of expression at blastoderm stage are under the control of the maternal pattern organizer gene nanos. In contrast, the expression of the second kni homologous gene is restricted to the late embryonic gonads. Due to its site of expression, we termed this gene 'embryonic gonad' (egon). In addition to the conserved DNA-binding domain, these three genes share an additional sequence of 19 amino acids, the kni-box, adjacent to the finger region. The identical N-terminal Cys/Cys finger encoded by each of the three genes suggests that they code for DNA-binding proteins which might bind to similar (or even identical) target sequences. PMID- 2555154 TI - Molecular cloning of human uracil-DNA glycosylase, a highly conserved DNA repair enzyme. AB - Uracil-DNA glycosylase is the DNA repair enzyme responsible for the removal of uracil from DNA, and it is present in all organisms investigated. Here we report on the cloning and sequencing of a cDNA encoding the human uracil-DNA glycosylase. The sequences of uracil-DNA glycosylases from yeast, Escherichia coli, herpes simplex virus type 1 and 2, and homologous genes from varicella zoster and Epstein-Barr viruses are known. It is shown in this report that the predicted amino acid sequence of the human uracil-DNA glycosylase shows a striking similarity to the other uracil-DNA glycosylases, ranging from 40.3 to 55.7% identical residues. The proteins of human and bacterial origin were unexpectedly found to be most closely related, 73.3% similarity when conservative amino acid substitutions were included. The similarity between the different uracil-DNA glycosylase genes is confined to several discrete boxes. These findings strongly indicate that uracil-DNA glycosylases from phylogenetically distant species are highly conserved. PMID- 2555155 TI - Reverse gyrase binding to DNA alters the double helix structure and produces single-strand cleavage in the absence of ATP. AB - Stoichiometric amounts of pure reverse gyrase, a type I topoisomerase from the archaebacterium Sulfolobus acidocaldarius were incubated at 75 degrees C with circular DNA containing a single-chain scission. After covalent closure by a thermophilic ligase and removal of bound protein molecules, negatively supercoiled DNA was produced. This finding, obtained in the absence of ATP, contrasts with the ATP-dependent positive supercoiling catalyzed by reverse gyrase and is interpreted as the result of enzyme binding to DNA at high temperature. Another consequence of reverse gyrase stoichiometric binding to DNA is the formation of a cleavable complex which results in the production of single strand breaks in the presence of detergent. Like eubacterial type I topoisomerase (protein omega), reverse gyrase is tightly attached to the 5' termini of the cleaved DNA. In the light of these results, a comparison is tentatively made between reverse gyrase and the eubacterial type I (omega) and type II (gyrase) topoisomerases. PMID- 2555156 TI - Transcriptional proofreading in Escherichia coli. AB - A novel transcriptional proofreading mechanism associated with the beta-subunit of wild-type RNA polymerase from Escherichia coli is suggested from the following data. The purified holoenzyme contains an NTPase activity which specifically converts noncognate NTPs to their corresponding NDP in a template-dependent manner during in vitro transcription of synthetic single- and double-stranded templates. In contrast, purified enzyme from an rpoB mutant which shows increased transcriptional error lacked template-dependent NTP hydrolytic activity. The NTP hydrolytic activity of wild-type enzyme was critically dependent on the integrity of the initiation complex, and required continued transcriptional elongation. Transcription and translation of the lacZ gene proceeded 17% faster in the mutant than in its wild-type parent. These results are discussed in terms of a proofreading model in which the rate of transcription is limited by proofreading events that involve recognition and hydrolysis of noncognate NTPs before they can be misincorporated into RNA. PMID- 2555157 TI - The putative transposase of transposable element Ac from Zea mays L. interacts with subterminal sequences of Ac. AB - The Ac-specific ORFa protein, overexpressed in a baculovirus system, specifically binds to several subterminal fragments of Ac. The 11 bp long inverted repeats of the transposable element are not bound by the ORFa protein. Major ORFa protein binding sites were delineated on 60 and 70 bp long sequence segments that lie 100 bp inside of the 5' Ac terminus and 40 bp inside of the 3' terminus respectively. Within all strongly bound fragments, and particularly in these 60 or 70 bp long segments, the hexamer motif AAACGG is repeated several times in direct or inverted orientation. The ORFa protein binds to synthetic concatemers of this motif, whereas the mutant motif AAAGGG is not complexed. Methylation of the cytosine residues in the AAACGG motif and/or its complementary strand has pronounced effects: whereas one of the two hemimethylated sequences has a higher affinity to the ORFa protein than both unmethylated and holomethylated DNAs, the other hemimethylated DNA is virtually not complexed at all. The native ORFa protein binding sites are more complex than the AAACGG sequence: certain Ac and Ds1 fragments devoid of AAACGG motifs (but containing several similar sequences) are weakly bound by the ORFa protein. PMID- 2555158 TI - Molecular basis of functional diversity of voltage-gated potassium channels in mammalian brain. AB - Cloning and sequencing of cDNAs isolated from a rat cortex cDNA library reveals that a gene family encodes several highly homologous K+ channel forming (RCK) proteins. Functional characterization of the channels expressed in Xenopus laevis oocytes following microinjection of in vitro transcribed RCK-specific RNAs shows that each of the RCK proteins forms K+ channels that differ greatly in both their functional and pharmacological properties. This suggests that the molecular basis for the diversity of voltage-gated K+ channels in mammalian brain is based, at least partly, on the expression of several RCK proteins by a family of genes and their assembly to homooligomeric K+ channels with different functional properties. PMID- 2555159 TI - Class II MHC molecules and the HIV gp 120 envelope protein interact with functionally distinct regions of the CD4 molecule. AB - A murine T cell hybridoma with a receptor specific for the class I molecule H-2 Dd was transfected with an expressible cDNA for human CD4. Expression of the human class II MHC molecule HLA-DP on Dd-positive murine fibroblasts resulted in a greatly enhanced response of the CD4-positive T cell hybridoma, measured either by lymphokine production or by rosette formation. Inhibition of these functional assays with anti-CD4 monoclonal antibodies implicated the two amino-terminal domains of CD4 in an interaction with the HLA-DP molecule. This interaction was blocked by incubation with recombinant gp120 envelope protein of HIV. In contrast, recombinant soluble CD4 did not inhibit and was able to prevent the inhibition by gp120. Anti-CD4 antibody blocking experiments clearly indicated that distinct regions of CD4 interact respectively with gp120 and with class II MHC molecules. PMID- 2555160 TI - Cell cycle-specific action of nerve growth factor in PC12 cells: differentiation without proliferation. AB - PC12 cells were manipulated in such a way as to permit the study of differentiation-specific responses independently from proliferative responses. Cells were starved for serum then exposed to nerve growth factor (NGF) or serum. Following addition of serum, cells incorporated thymidine in a synchronous manner. Subsequent to the wave of DNA synthesis, the cell number increased approximately two-fold. Addition of NGF to serum-starved cultures had no measurable effect on either parameter. Neurite outgrowth was more rapid and extensive and appearance of Na+ channels, measured as saxitoxin binding sites, more rapid than when NGF was added to exponentially-growing cells. Epidermal growth factor receptors were heterologously down-regulated by NGF with similar kinetics under both conditions. Induction of the proto-oncogene c-fos by NGF was also greater in the serum-starved cells than in exponentially-growing cultures. These results indicated that serum starvation resulted in synchronisation of the cultures and that NGF action may be cell cycle-specific. Analysis of the cellular response to NGF at different times during the cell cycle showed that c-fos was induced in the G1 phase but not in S or G2. Fluorescence-activated cell sorter analysis demonstrated that addition of NGF to exponentially-growing cells, resulted in their accumulation in a G1-like state. With regard to the study of the mechanism of NGF action, these results illustrate that measurements of NGF effects on specific components in the signal transduction pathway may be confounded by the use of exponentially-growing cultures. PMID- 2555161 TI - Structure, mapping and expression of a growth factor inducible gene encoding a putative nuclear hormonal binding receptor. AB - We have characterized a growth factor inducible gene, N10, encoding a nuclear protein of 601 amino acids with a significant similarity to members of the steroid and thyroid hormone receptor families. The gene is rapidly but transiently induced by several mitogens. Immunoprecipitation studies show that the N10 protein is transiently expressed after stimulation of quiescent cells, presenting a half-life of approximately 30 min. The N10 transcription unit is 8 kb in length, split into seven exons. The exon-intron distribution is in general similar to that of other members of the nuclear receptor superfamily, but presents some differences which suggest that N10 belongs to a new family of these molecules. The 5' flanking region contains one DSE which could explain its immediate response to external stimulus. The N10 gene is located in the [F1-F3] region of mouse chromosome 15. PMID- 2555162 TI - Phospholipase C-gamma, a substrate for PDGF receptor kinase, is not phosphorylated on tyrosine during the mitogenic response to CSF-1. AB - Quiescent mouse NIH3T3 cells expressing a transduced human c-fms gene encoding the receptor for colony stimulating factor-1 (CSF-1) were stimulated with mitogenic concentrations of platelet-derived growth factor (PDGF) or CSF-1. Immunoprecipitated phospholipase C-gamma (PLC-gamma) was phosphorylated on tyrosine and calcium was mobilized following treatment of intact cells with PDGF. In contrast, only trace amounts of phosphotyrosine were incorporated into PLC gamma and no intracellular calcium signal was detected after CSF-1 stimulation. Similarly, CSF-1 treatment did not stimulate phosphorylation of PLC-gamma on tyrosine in a CSF-1-dependent. SV40-immortalized mouse macrophage cell line that expresses high levels of the CSF-1 receptor. In fibroblasts, antiserum to PLC gamma co-precipitated a fraction of the tyrosine phosphorylated form of the PDGF receptor (PDGF-R) after ligand stimulation, implying that phosphorylated PDGF-R and PLC-gamma were associated in a stable complex. Pre-treatment of cells with orthovanadate also led to tyrosine phosphorylation of PLC-gamma which was significantly enhanced by PDGF, but not by CSF-1. Thus, although the PDGF and CSF 1 receptors are structurally related and appear to be derived from a single ancestor gene, only PDGF-induced mitogenesis in fibroblasts correlated with tyrosine phosphorylation of PLC-gamma. PMID- 2555163 TI - PEA3 is a nuclear target for transcription activation by non-nuclear oncogenes. AB - We have found that the activity of the transcription factor PEA3 is regulated by the expression of non-nuclear oncogenes. This factor, although distinct from PEA1 (AP1), is activated by the same oncogenes (v-src, polyoma (Py) middle T, c-Ha ras, v-mos, v-raf), by tetradecanoyl phorbol-acetate (TPA) and by serum components. We present evidence that PEA3 and PEA1 co-operate in the response of the polyoma virus (Py) alpha domain to oncogene expression. However, in contrast to PEA1, c-fos does not appear to be necessary for activation of PEA3, suggesting that PEA3 is a fos independent target for regulation of transcription by non nuclear oncogenes. PMID- 2555164 TI - MyoD expression in the forming somites is an early response to mesoderm induction in Xenopus embryos. AB - We describe the cloning, cDNA sequence and embryonic expression of a Xenopus homologue of MyoD, a mouse gene encoding a DNA-binding protein that can activate muscle gene expression in cultured cells. The predicted Xenopus MyoD protein sequence is remarkably similar to mouse MyoD. Zygotic expression of MyoD begins in early gastrulae, but there is a low level of unlocalized maternal message. Northern blot analysis of dissected embryos and in situ hybridization show that MyoD RNA is restricted to the gastrula mesoderm and to the somites of neurulae and tailbud embryos. The time and place of MyoD expression are consistent with a role for MyoD in the activation of other muscle genes in the somites of the frog embryo. However, MyoD is skeletal muscle-specific and is not expressed even in the early embryonic heart, which co-expresses cardiac and skeletal actin isoforms. Striated muscle genes can therefore be activated in some embryonic tissues in the absence of MyoD. The concentration of MyoD in the somites falls once they have formed, suggesting that MyoD may act there transiently to establish muscle gene expression. MyoD transcription is activated following mesoderm induction, and is the earliest muscle-specific response to mesoderm inducing factors so far described. PMID- 2555165 TI - A defective proton pump, point-mutated bacteriorhodopsin Asp96----Asn is fully reactivated by azide. AB - Addition of azide fully restored the proton pump activity of defective bacteriorhodopsin (BR) mutant protein Asp96----Asn. The decay time of M of BR Asp96----Asn, the longest living intermediate, was decreased from 500 ms at pH 7.0 to approximately 1 ms under conditions of saturating azide concentrations. This decay was faster than the decay of M in the wild-type, where no such azide effect was detectable. Stationary photocurrents, measured with purple membranes immobilized and oriented in a polyacrylamide gel, increased upon addition of azide up to the level of the wild-type. Different small anions of weak acids restored the pump activity with decreasing affinity in the order: cyanate greater than azide greater than nitrite greater than formiate greater than acetate. The activation energy of the M decay in the mutant was higher in the presence (48 kJ/mol) than in the absence (27 kJ/mol) of 100 mM azide even though the absolute rate was dramatically increased by azide. This effect of azide is due to the substitution of a carboxamido group for a carboxylic group at position 96 which removes the internal proton donor and causes an increase in the entropy change of activation for proton transfer which is reversed by azide. PMID- 2555166 TI - Action at a distance in Mu DNA transposition: an enhancer-like element is the site of action of supercoiling relief activity by integration host factor (IHF). AB - The first committed step in the in vitro strand transfer reaction of a mini-Mu donor molecule is the formation of a Type 1 complex in which the Mu ends are held together in a non-covalent protein-DNA complex. Efficient formation of this complex at high levels of donor supercoiling (sigma approximately -0.06) requires the Mu A and Escherichia coli HU proteins. At in vivo levels of supercoiling, efficient reaction also requires E. coli integration host factor (IHF). We demonstrate that this supercoiling relief activity of IHF is mediated through an IHF binding site in the Mu early promoter region. This site is part of a larger enhancer-like element which includes operator 1 (01) and part of operator 2 (02) with the IHF site in between. The enhancer-like element stimulates the initial rate of the in vitro reaction 100-fold and acts in a distance-independent fashion. Inversion of the orientation of the element results in a total loss of enhancer activity in the absence of IHF. However, a 10-fold stimulation in the initial rate of reaction is induced by the addition of IHF. Furthermore, correct helical phasing between 01 and 02 is required for maximal activity. The results indicate that a specific geometrical configuration of the enhancer-like element, which includes a sharp bend between 01 and 02, is required for optimal induction of synapsis. PMID- 2555167 TI - The extreme mutator effect of Escherichia coli mutD5 results from saturation of mismatch repair by excessive DNA replication errors. AB - Escherichia coli mutator mutD5 is the most potent mutator known. The mutD5 mutation resides in the dnaQ gene encoding the proofreading exonuclease of DNA polymerase III holoenzyme. It has recently been shown that the extreme mutability of this strain results, in addition to a proofreading defect, from a defect in mutH, L, S-encoded postreplicational DNA mismatch repair. The following measurements of the mismatch-repair capacity of mutD5 cells demonstrate that this mismatch-repair defect is not structural, but transient. mutD5 cells in early log phase are as deficient in mismatch repair as mutL cells, but they become as proficient as wild-type cells in late log phase. Second, arrest of chromosomal replication in a mutD5-dnaA(Ts) strain at a nonpermissive temperature restores mismatch repair, even from the early log phase of growth. Third, transformation of mutD5 strains with multicopy plasmids expressing the mutH or mutL gene restores mismatch repair, even in rapidly growing cells. These observations suggest that the mismatch-repair deficiency of mutD strains results from a saturation of the mutHLS-mismatch-repair system by an excess of primary DNA replication errors due to the proofreading defect. PMID- 2555168 TI - Heteroduplex substrates for bacteriophage lambda site-specific recombination: cleavage and strand transfer products. AB - Lambda's Int protein acts as a specific topoisomerase at attachment sites, the DNA segments that are required for site-specific recombination. Int cleaves each strand of an attachment site at a unique place and creates strand exchanges by joining broken ends from two different parents. To study the action of Int topoisomerase in more detail, heteroduplex attachment sites were made by annealing strands that are complementary except for a few base pairs that lie in the region between the points of top and bottom strand exchange in the attachment site core. These heteroduplexes appear to interact normally with Int and its accessory proteins IHF and Xis. Although the heteroduplex sites are specifically cleaved by Int topoisomerase, rejoining of the broken DNA is hindered by the lack of Watson--Crick complementarity adjacent to the break. Because of this, heteroduplexes accumulate broken intermediates which are then processed in novel ways. We have used this feature to provide new information about functional differences between attachment sites, to investigate the way Xis protein controls directionality of site-specific recombination, and to demonstrate that Int protein can join strands indiscriminately and can therefore generate recombinants with either of two genetic polarities. PMID- 2555169 TI - Deletion of the gene for subunit III leads to defective assembly of bacterial cytochrome oxidase. AB - COIII is one of the major subunits in the mitochondrial and a bacterial cytochrome c oxidase, cytochrome aa3. It does not contain any of the enzyme's redox-active metal centres and can be removed from the enzyme without major changes in its established functions. We have deleted the COIII gene from Paracoccus denitrificans. The mutant still expresses spectroscopically detectable enzyme almost as the wild-type, but its cytochrome c oxidase activity is much lower. From 50 to 80% of cytochrome a is reduced and its absorption maximum is 2 3 nm blue-shifted. The EPR signal of ferric cytochrome a is heterogeneous indicating the presence of multiple cytochrome a species. Proteolysis of the membrane-bound oxidase shows new cleavage sites both in COI and COII. DEAE chromatography of solubilized enzyme yields fractions that contain a COI + COII complex and in addition haem-binding, free COI as well as free COII. The mutant phenotype can be complemented by introducing the COIII gene back to cells in a plasmid vector. We conclude that cytochrome oxidase assembles inefficiently in the absence of COIII and that this subunit may facilitate a late step in the assembly. The different oxidase species in the mutant represent either accumulating intermediates of the assembly pathway or dissociation products of a labile COI + COII complex and its conformational variants. PMID- 2555170 TI - Differential regulation of three sodium channel messenger RNAs in the rat central nervous system during development. AB - The levels of the mRNAs encoding sodium channels I, II and III in various regions of the developing rat central nervous system (from embryonal day 10 to postnatal day 90) have been examined by blot hybridization analysis with specific probes. The three sodium channel mRNAs exhibit different temporal and regional expression patterns. The expression of sodium channel I mRNA rises after a lag phase to adult levels during the second and third postnatal weeks with stronger increases in caudal regions of the brain and in spinal cord. Sodium channel II mRNA increases steadily until the first postnatal week, keeping high adult levels in rostral regions of the brain or reaching low adult levels after the second postnatal week in most caudal regions of the brain and in spinal cord; cerebellum shows low levels during the first two postnatal weeks but high adult levels. In all regions, sodium channel III mRNA attains maximum levels around birth and decreases during the first and second postnatal weeks to reach variable low adult levels. These results suggest that sodium channel III is expressed predominantly at fetal and early postnatal stages and sodium channel I predominantly at late postnatal stages, whereas sodium channel II is expressed throughout the developmental stages studied with greater regional variability. PMID- 2555171 TI - Expression cloning of a receptor for human granulocyte-macrophage colony stimulating factor. AB - Two cDNA clones encoding a receptor for human granulocyte-macrophage colony stimulating factor (hGM-CSF-R) were isolated by expression screening of a library made from human placental mRNA. Pools of recombinant plasmid DNA were electroporated into COS cells which were then screened for their capacity to bind radioiodinated hGM-CSF using a sensitive microscopic autoradiographic approach. The cloned GM-CSF-R precursor is a 400 amino acid polypeptide (Mr 45,000) with a single transmembrane domain, a glycosylated extracellular domain and a short (54 amino acids) intracytoplasmic tail. It does not contain a tyrosine kinase domain nor show homology with members of the immunoglobulin super gene family, but does show some significant sequence homologies with receptors for several other haemopoietic growth factors, including those for interleukin-6, erythropoietin and interleukin-2 (beta-chain) and also to the prolactin receptor. When transfected into COS cells the cloned cDNA directed the expression of a GM-CSF-R showing a single class of affinity (KD = 2(-8) nM) and specificity for human GM CSF but not interleukin-3. Messenger RNA coding for this receptor was detected in a variety of haemopoietic cells known to display hGM-CSF binding, and cross linking experiments revealed a similar size for the glycosylated receptors in transfected COS and haemopoietic cells. PMID- 2555172 TI - trkB, a novel tyrosine protein kinase receptor expressed during mouse neural development. AB - We have isolated a novel member of the tyrosine protein kinase family of cell surface receptors. This gene, designated trkB, is highly related to the human trk proto-oncogene. At the amino acid level, their respective products share a 57% homology in their extracellular regions including 9 of the 11 cysteines present in the trk proto-oncogene. This homology increases to 88% within their respective tyrosine kinase catalytic domains. Both trk and trkB are equally distantly related to the other members of this gene family of receptors. A biologically active cDNA clone of trkB can direct the synthesis of gp145trkB, a glycoprotein of 145 kd of which only 93 kd correspond to its polypeptide backbone. In adult mice, trkB is preferentially expressed in brain tissue, although significant levels of trkB RNA have also been observed in lung, muscle and ovaries. In addition, trkB transcripts can be detected in mid and late gestation embryos. The trkB locus exhibits a complex pattern of transcription. At least seven RNA species ranging in size from approximately 9 kb to 2 kb have been identified in brain. However, only a subset of these transcripts appears to be expressed in the other tissues. In situ hybridization analysis of 14 and 18 day old mouse embryos indicates that trkB transcripts are localized in the central (CNS) and peripheral (PNS) nervous systems, including brain, spinal cord, spinal and cranial ganglia, paravertebral trunk of the sympathetic nervous system and various innervation pathways. These results suggest that trkB may code for a novel cell surface receptor involved in neurogenesis. PMID- 2555173 TI - Dual control of C-reactive protein gene expression by interleukin-1 and interleukin-6. AB - Human C-reactive protein (CRP) is the major acute phase reactant during acute inflammation. The human CRP promoter is expressed in an inducible and cell specific manner when linked to the bacterial CAT gene and transfected into human hepatoma cell cultures. In this paper we analyze the effect of several recombinant cytokines or CRP promoter inducibility in human Hep3B cells. When cytokines are tested singly the major inducer of CRP-CAT fusions is interleukin-6 (IL-6). Maximal CAT gene expression, however, is only achieved when both interleukin-1 beta (IL-1 beta) and IL-6 are present. The response to the two cytokines is cooperative. Cooperativity is maintained when the CRP promoter is linked to a different coding region, that of the bacterial neomycin phosphotransferase II gene. With a series of 5' and 3' deletions we show the existence of two distinct and independent regions responsive to IL-6 and located upstream to the TATA box. The IL-1 effect is exerted at the level of downstream sequences that are probably important for optimal mRNA translatability or nuclear cytoplasmic transport. Inducibility is not influenced by the activation of protein kinases C or A and does not require new protein synthesis. PMID- 2555174 TI - TNF stimulates expression of mouse MHC class I genes by inducing an NF kappa B like enhancer binding activity which displaces constitutive factors. AB - We have dissected the mouse H-2Kb gene promoter in order to define the sequences responsible for induction by tumour necrosis factor (TNF-alpha). An enhancer element (-187 to -158) composed of two imperfect direct palindromic repeats has been shown to be necessary and sufficient for TNF-alpha induction of a heterologous promoter. A multimer of either repeat is also responsive, while a single copy is not: this is the situation in the beta 2-microglobulin (beta 2-m) promoter which contains a single palindrome and does not respond to TNF-alpha. We had previously found that the two repeats can bind a factor named KBF1. We show here that in the uninduced state the transcription factor AP2 binds to the interpalindromic region, while in TNF-treated cells an NF kappa B-like activity is induced which displaces both KBF1 and AP2 and binds to the two palindromes. This strongly suggests that induction of an NF kappa B-like activity is responsible for TNF-alpha stimulation of mouse MHC class I genes. PMID- 2555175 TI - Identification of four conserved motifs among the RNA-dependent polymerase encoding elements. AB - Four consensus sequences are conserved with the same linear arrangement in RNA dependent DNA polymerases encoded by retroid elements and in RNA-dependent RNA polymerases encoded by plus-, minus- and double-strand RNA viruses. One of these motifs corresponds to the YGDD span previously described by Kamer and Argos (1984). These consensus sequences altogether lead to 4 strictly and 18 conservatively maintained amino acids embedded in a large domain of 120 to 210 amino acids. As judged from secondary structure predictions, each of the 4 motifs, which may cooperate to form a well-ordered domain, places one invariant amino acid in or proximal to turn structures that may be crucial for their correct positioning in a catalytic process. We suggest that this domain may constitute a prerequisite 'polymerase module' implicated in template seating and polymerase activity. At the evolutionary level, the sequence similarities, gap distribution and distances between each motif strongly suggest that the ancestral polymerase module was encoded by an individual genetic element which was most closely related to the plus-strand RNA viruses and the non-viral retroposons. This polymerase module gene may have subsequently propagated in the viral kingdom by distinct gene set recombination events leading to the wide viral variety observed today. PMID- 2555176 TI - Activation of SV40 DNA replication in vitro by cellular protein phosphatase 2A. AB - We have made use of the cell-free SV40 DNA replication system to identify and characterize cellular proteins required for efficient DNA synthesis. One such protein, replication protein C (RP-C), was shown to be involved with SV40 large T antigen in the early stages of viral DNA replication in vitro. We demonstrate here that RP-C is identical to the catalytic subunit of cellular protein phosphatase 2A (PP2Ac). The purified protein dephosphorylates specific phosphoamino acid residues in T antigen, consistent with the hypothesis that SV40 DNA replication is regulated by modulating the phosphorylation state of the viral initiator protein. We also show that purified RP-C/PP2Ac preferentially stimulates SV40 DNA replication in extracts from early G1 phase cells. This finding suggests that the activity of a cellular factor that influences the net phosphorylation state of T antigen is cell cycle dependent. PMID- 2555177 TI - Disruption of the yeast nuclear PET54 gene blocks excision of mitochondrial intron aI5 beta from pre-mRNA for cytochrome c oxidase subunit I. AB - The nuclear PET54 gene of Saccharomyces cerevisiae was cloned and a pet54::LEU2 gene disruption strain was constructed. Analysis of the phenotype of this strain revealed a defect in expression of two mitochondrial genes: COX1, which encodes cytochrome c oxidase subunit I, and COX3, which encodes cytochrome c oxidase subunit III. The defect in COX1 gene expression in the pet54 mutant was shown to be the result of inefficient excision of COX1 intron aI5 beta. Two lines of evidence indicate that inefficient excision of intron aI5 beta is the sole defect in COX1 gene expression. First, a pet54::LEU2 cytoductant bearing the 'short' mitochondrial genome that lacks both COX1 introns aI5 alpha and aI5 beta is defective only in COX3 gene expression and not in COX1 mRNA splicing or mRNA translation. Second, Northern analysis of COX1 transcipts from the pet54 mutant showed that a 3.8 kb COX1 transcript containing unexcised intron aI5 beta and lacking intron aI5 alpha is accumulated while the amount of 2.2 kb mature COX1 mRNA is diminished. In an effort to relate the role of the PET54 gene product in splicing of COX1 pre-mRNA to the previously characterized role for PET54 in translation of mitochondrial COX3 mRNA, the sequence of the PET54-responsive portion of the COX3 5' untranslated leader region was compared to the COX1 intron aI5 beta sequence. Two blocks of RNA sequence present in COX3 have similar counterparts within intron aI5 beta of COX1. The possibility that the PET54 protein binds to one or the other of these blocks of RNA sequence and the potential consequences of this interaction are discussed. PMID- 2555178 TI - HPV16 E6 and E7 proteins cooperate to immortalize human foreskin keratinocytes. AB - The human papillomavirus types (HPVs) most often associated with cancer of the cervix, such as HPV16, have been reported previously to immortalize normal human foreskin keratinocytes in vitro, while the types that are primarily associated with benign cervical lesions failed to do so. In this study we have determined the HPV16 genes that are responsible for the immortalizing activity of the viral genome. Transfection with a plasmid in which E6 and E7 were the only intact open reading frames (ORFs) induced an indefinite life-span in the keratinocytes with an efficiency similar to that of the entire early region of the viral DNA. Mutants in the E6E7 clone with inactivating lesions in E6 or E7 failed to induce immortalization. When transfected alone, E7 could induce hyperproliferation, but these cells eventually senesced. By itself, E6 exhibited no activity, Co transfection of a plasmid with an intact E6 ORF and a second plasmid with an intact E7 ORF generated keratinocyte lines with indefinite growth potential. The E6 and E7 proteins were detected in the lines induced by the E6E7 DNA and by co transfection of the E6 and E7 plasmids. Therefore, we conclude that HPV16 E6 and E7 cooperative to immortalize human keratinocytes in vitro. Changes in cellular gene expression are probably also required for immortalization since all of the keratinocyte lines examined were aneuploid. Serum and calcium resistant sublines were isolated from the E6E7 induced lines, indicating that other HPV genes do not play an obligatory role in the generation of resistance to differentiation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555179 TI - Molecular cloning and structural analysis of a gene from Zea mays (L.) coding for a putative receptor for the plant hormone auxin. AB - The major auxin-binding protein from maize coleoptiles was purified to homogeneity. The protein has an apparent mol. wt of 22 kd and binds 1 naphthylacetic acid with a KD of 2.40 x 10(-7) M. Additional antigenically related proteins, present in very low amounts, could be demonstrated in maize coleoptiles using immunodetection. Extensive protein sequence analysis of the major auxin-binding protein allowed the construction of several synthetic oligonucleotide probes which were used to isolate a cDNA coding for this protein. The cDNA corresponds to a mRNA with a 3'-poly(A)+ sequence and a single, long open reading frame of 603 bases. The open reading frame, starting 34 residues from the 5' end of the cDNA, predicts a 21,990 Dalton protein of 201 amino acids. Comparison of this deduced amino acid sequence with the partial amino acid sequences of purified auxin-binding protein, revealed a perfect match, involving a total of 53 amino acid residues. The primary amino acid sequence includes a 38 amino-acid-long N-terminal hydrophobic leader sequence which could represent a signal for translocation of this protein to the endoplasmic reticulum. An additional signal is located at the C-terminal end, consisting of the amino acids KDEL known to be responsible for preventing secretion of proteins from the lumen of the endoplasmic reticulum in eucaryotic cells. The primary sequence contains a N-glycosylation site (-asp133-thr-thr-). This site was found to be glycosylated by a high-mannose-type oligosaccharide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555180 TI - cDNA clones of the auxin-binding protein from corn coleoptiles (Zea mays L.): isolation and characterization by immunological methods. AB - An auxin-binding protein (ABP) cDNA clone was selected from a lambda gt11 cDNA library from corn coleoptiles with highly purified IgGanti ABP. The sequence of 794 bp contains an open reading frame (ORF) of 603 bp, coding for a 22 kd protein. There are indications of a signal peptide of 38 amino acids (von Heijne, G. 1983, Eur. J. Biochem., 133, 17-21). A N-glycosylation site can be deduced and a C-terminal KDEL amino acid sequence is detected. An EcoRI fragment containing the beginning portion of the cDNA with about three quarters of the ORF was used to select cDNA clones from an independently produced lambda gt11 cDNA library of corn coleoptiles. Northern blot analysis with in vitro transcribed biotinylated RNA showed a single band of not more than 850 bases. The full-length in vitro transcript directed the in vitro synthesis of a protein which is precipitated by IgGanti ABP. Rabbit antibodies raised against a fusion protein detect the ABP as a double band on Western blots. Only the smaller of the two ABP bands is labeled by two different KDEL-specific IgG preparations. PMID- 2555181 TI - Isolation of cDNA clones encoding human acid sphingomyelinase: occurrence of alternatively processed transcripts. AB - Acid sphingomyelinase (sphingomyelin phosphodiesterase, EC 3.1.4.12) was purified from human urine and 12 tryptic peptides were microsequenced (128 residues). Based on regions of minimal codon redundancy, four oligonucleotide mixtures were synthesized and oligonucleotide mixture 1 (20mer; 256 mix) was used to screen 3 X 10(6) independent recombinants from a human fibroblast cDNA library. Putative positive clones (92) were purified and analyzed by Southern hybridization with oligonucleotide mixtures 2-4. These studies revealed two groups of clones; group 1 (80 clones; inserts ranging from approximately 1.2 to 1.6 kb) hybridized with oligonucleotides mixtures 1-4, while group II (12 clones; inserts ranging from approximately 1.2 to 1.4 kb) hybridized with oligonucleotide mixtures 1-3. Several group II clones had larger inserts than those in group I, but did not hybridize with oligonucleotide mixture 4. Screening of a human placental cDNA library with a 450 bp group I fragment, also resulted in the isolation of group I and II clones. Representative clones from group I (pASM-1) and group II (pASM-2) were sequenced. pASM-1 contained a 1879 bp insert which was colinear with 96 microsequenced amino acids, while the pASM-2 1382 bp insert was colinear with 78 microsequenced residues. Notably, pASM-2 did not have an internal 172 bp sequence encoding 57 amino acid residues, but had instead an in-frame 40 bp sequence encoding 13 amino acids which was not present in pASM-1. These findings demonstrate the presence of two distinct acid sphingomyelinase transcripts in human fibroblasts and placenta and suggest the occurrence of alternative processing of the mRNA encoding this lysosomal hydrolase. PMID- 2555182 TI - Reversible abrogation of IL-3 dependence by an inducible H-ras oncogene. AB - Immortalized, interleukin-3 (IL-3)-dependent mouse mast cells (PB-3c) were transfected with a human activated c-H-ras gene under the transcriptional control of the mouse mammary tumor virus long terminal repeat. Addition of increasing amounts of dexamethasone resulted in a concentration-dependent increase in expression of the H-ras oncogene. The elevation of p21 ras protein concentrations was paralleled by progressive growth of the transfectants in the absence of exogenous IL-3, leading to complete abrogation of growth-factor requirement at high p21ras levels. The maintenance of the IL-3-independent state required the continuous expression of the H-ras oncogene, since dexamethasone removal was followed by rapid cell death. Expression of the H-ras oncogene induced PB-3c cells to produce IL-3 and granulocyte-macrophage colony-stimulating factor, suggesting that their IL-3-independent proliferation may be due to an autocrine mechanism. PMID- 2555183 TI - Role of myristoylation of poliovirus capsid protein VP4 as determined by site directed mutagenesis of its N-terminal sequence. AB - Mutations were introduced by oligonucleotide-directed mutagenesis into the cDNA of poliovirus type 1 (Mahoney) in the region coding for the first five amino acids (myristoylation signal) of the viral capsid protein precursor P1. The cDNAs were then transcribed in vitro and the properties of the transcripts carrying the mutations studied in vitro by translation in a reticulocyte lysate or in vivo upon transfection of primate cells. Mutation of amino acid residue number 5 (Ser5 ---Thr) did not affect the viral phenotype, whereas mutations of residues number 1 (Gly1----Arg), 2 (Ala2----Pro) or 5 (Ser5----Pro) prevented myristoylation of P1 and were lethal. However, delayed production of virus was occasionally observed as the result of reverse mutations, which were found to restore a functional myristoylation signal as well as a wild-type virus phenotype. Thus, the myristoylation signal of the poliovirus polyprotein can accommodate Ala, Ser, Thr or Leu residues at position 2 and Ser, Thr or Ala residues at position 5. Mutations that altered myristoylation of P1 and affected virus viability did not prevent replication of the viral RNA but severely impeded in vitro processing of P1. This suggests that myristoylation plays a role in poliovirus capsid protein assembly. PMID- 2555184 TI - A cinematographic view of Escherichia coli RNA polymerase translocation. AB - A series of RNA synthesizing transcription complexes, initiated at the T7 A1 promoter and halted at specific base positions ranging from +12 to +40, were analyzed by footprinting techniques; exonuclease III was used to determine the position of the bound RNA polymerase on the DNA and hydroxyl radicals were used to visualize the protein--DNA contact sites within the protected areas. In the binding (open) complex without RNA there are two DNA-domains, differing in their protection pattern. The first, extending from position +18 to -13, termed 'melting domain', is fully protected, whereas the second, extending from -14 to 55, termed 'recognition domain', shows only partial protection. At this domain, RNA polymerase is attached to one side of the DNA only, as indicated by the 10-bp periodicity of the protection pattern. Our data show that the formation of a mature RNA transcribing complex is characterized by dissociation of the RNA polymerase from the recognition domain, whereby the size of the melting domain remains constant. This process is accomplished if the nascent RNA has reached a length of 11 bases. As the RNA reaches a length of 20 bases, the size of the melting domain decreases from approximately 30 to 23 bp. Further RNA synthesis leaves the protection pattern essentially unchanged. These data demonstrate that the formation of a mature RNA transcribing complex can be described by at least two transitions. PMID- 2555185 TI - Bordetella pertussis adenylate cyclase: purification and characterization of the toxic form of the enzyme. AB - Bordetella pertussis produces a calmodulin-sensitive adenylate cyclase (AC) which is an essential virulence factor in mammalian pertussis. Here we report the purification and characterization of the toxic form of the enzyme, which penetrates eukaryotic cells and generates high levels of intracellular cAMP. This form was purified from an extract of B.pertussis strain carrying a recombinant plasmid which over-produced both enzymatic and toxic activities of the enzyme. Western blot analysis of the extract using anti-B.pertussis AC antibodies detected only one protein of 200 kd. However, gel filtration of the extract resolved two peaks of enzymatic activity. The first peak of aggregated material contained greater than 70% of the total enzymatic activity, and the second peak contained the majority of the toxic activity. Purification of the enzyme from both peaks yielded proteins of 200 kd, with similar biochemical and immunological properties. Yet only the enzyme purified from the second peak could penetrate human lymphocyte and catalyse the formation of intracellular cAMP. B.pertussis AC gene expressed in Escherichia coli produced a calmodulin-dependent enzyme of 200 kd, which lacked lymphocyte penetration capacity. It is proposed that a post translational modification that occurs in B.pertussis but not in E.coli confers upon the 200 kd protein of B.pertussis AC the toxic properties. PMID- 2555186 TI - Characterization of the staphylococcal beta-lactamase transposon Tn552. AB - The staphylococcal beta-lactamase transposon Tn552 is a member of a novel group of transposable elements. The organization of genes in Tn552 resembles that of members of the Tn21 sub-group of Tn3 family transposons, which transpose replicatively by cointegrate formation and resolution. Thus, a possible resolution site ('resL') and a resolvase gene (tnpR or 'binL') have been identified. However, consistent with the fact that Tn552 generates 6 bp (rather than 5 bp) flanking direct repeats of target DNA, neither the putative transposase protein, nor the terminal inverted repeats of Tn552 are homologous to those of Tn3 elements. Tn552, like phage Mu and retroelements, is defined by the terminal dinucleotides 5' TG .. CA 3'. A naturally occurring staphylococcal plasmid, pI9789, contains a Tn552-derived resolution system ('resR-binR') that acts as a 'hotspot' for Tn552 transposition; insertion creates a segment of DNA flanked by inversely repeated resolution sites, one (resR) on pI9789 and the other (resL) on Tn552. The putative Tn552 resolvase, the most closely related of known resolvases to the homologous DNA invertases, initially was identified as a DNA invertase ('Bin') as a result of its ability to mediate efficient inversion of this segment in vivo. PMID- 2555187 TI - Carotid baroreflex sensitivity at rest and during exercise is not influenced by opioid receptor antagonism. AB - Physical effort involves, along with an increase in the plasma concentration of beta-endorphin, profound cardiovascular adaptations. The aim of the present study was to investigate with the use of the variable neck chamber technique, the influence of the endogenous opioids on the carotid baroreflex control of blood pressure and heart rate at rest as well as during exercise. Ten normal volunteers exercised in the supine position up to 33% and 66% of their maximal exercise capacity and received, in a randomized double-blind cross-over protocol, either saline or naloxone (10 mg intravenously, followed by a continuous infusion of 10 mg.h-1). During exercise a progressive attenuation of the carotid baroreceptor reflex control of blood pressure and heart rate was noted. However, neither at rest nor during exercise, did opioid antagonism influence the carotid baroreceptor control of blood pressure and heart rate. Intra-arterial pressure and heart rate also remained unaffected. In contrast, both at rest and during exercise, naloxone administration produced a significant increase in the plasma concentration of cortisol. The latter suggests that in vivo the opioid receptors were effectively antagonized. In conclusion the present study confirms that opioids play only a minor role in cardiovascular homeostasis at rest. In addition, this study demonstrates that they are not involved in the cardiovascular adaptation to exercise, nor in the exercise-related attenuation of the carotid baroreceptor control of pressure and heart rate. PMID- 2555188 TI - Influence of endogenous opioids on atrial natriuretic factor release during exercise in man. AB - To evaluate to what extent opioid secretion in exercise induces the release of atrial natriuretic factor (ANF), six healthy male volunteers who were trained subjects, were submitted to two maximal exercise tests with and without (control) opioid receptor blockade by Naltrexone. Blood samples were drawn before (rest) and after exercise (post-exercise) in order to measure human ANF (alpha h ANF), beta-endorphin, plasma aldosterone concentration (PAC) plasma renin activity (PRA) and adreno-cortico trophic hormone (ATCH) by radio-immunological methods. Expired gas was collected during exercise to measure oxygen consumption. On average, the same maximal oxygen consumption (VO2max) during exercise was reached by all subjects with and without treatment. Plasma ANF level at rest slightly decreased after administration of Naltrexone; the response to physical exercise was significantly reduced by Naltrexone. There was no statistical difference between plasma levels of beta-endorphin, PRA and ACTH at rest nor in the post exercise situation under the influence of Naltrexone. The PAC increased significantly at rest after Naltrexone administration but there was no statistical difference between both values after exercise. These data demonstrate that: (1) ANF secretion during exercise is influenced by the level of beta endorphin in the plasma; (2) the possible inhibitory role of ANF on aldosterone secretion during exercise is probably over-ruled by the increase in plasma ACTH and PRA. PMID- 2555189 TI - Role of the cell surface-exposed regions of outer membrane protein PhoE of Escherichia coli K12 in the biogenesis of the protein. AB - To investigate the role of the cell surface-exposed regions of outer membrane protein PhoE of Escherichia coli K12 in the biogenesis of the protein, deletions were generated in two presumed cell surface-exposed regions of the protein. Intact cells expressing these mutant proteins were recognized by PhoE-specific monoclonal antibodies, which recognize conformational epitopes on the cell surface-exposed parts of the protein and/or were sensitive to a PhoE-specific phage. This shows that the polypeptides were normally incorporated into the outer membrane. When the deletions extended four amino acid residues into the seventh presumed membrane-spanning segment, the polypeptides accumulated in the periplasm. In conclusion, exposed regions of PhoE protein apparently do not play an essential role in outer membrane localization, which is consistent with the observation that these regions are hypervariable when PhoE is compared to the related proteins OmpF and OmpC. In contrast, the membrane-spanning segments are essential for the assembly process. PMID- 2555190 TI - Concerted action of cosolvents, chaotropic anions and thioredoxin on chloroplast fructose-1,6-bisphosphatase. Reactivity to iodoacetamide. AB - The incubation of chloroplast fructose-1,6-bisphosphatase with both dithiothreitol and protein denaturants made sulfhydryl groups available for reaction with [1-14C]iodoacetamide (10-12 mol iodoacetamide incorporated/mol enzyme). Digestion of S-carboxyamidomethylated enzyme with trypsin and polyacrylamide gel electrophoresis, in the presence of sodium dodecylsulfate, yielded two 14C-labeled fragments whose apparent molecular mass were 10 kDa and 16 kDa. In the absence of either dithiothreitol or protein denaturants the incorporation of iodoacetamide to the enzyme was lower than 4 mol. When chloroplast fructose-1,6-bisphosphatase was initially incubated with dithiothreitol (2.5 mM) and (a) high concentrations of both fructose 1,6 bisphosphate (4 mM) and Ca2+ (0.3 mM) or (b) low concentrations of both fructose 1,6-bisphosphate (0.8 mM) and Ca2+ (0.05 mM) in the presence of either 2-propanol (15%, by vol.), trichloroacetate (0.15 M) or chloroplast thioredoxin-f (0.5 microM) and subsequently subjected to proteolysis and electrophoresis, S carboxyamidomethylated tryptic fragments had similar molecular masses. Thus, conditions that stimulated the specific activity of chloroplast fructose-1,6 bisphosphatase caused conformational changes which favoured both the reduction of disulfide bridges and the exposure of sulfhydryl groups. In this aspect, thioredoxin exerted structural and kinetic effects similar to compounds not involved in redox reactions (organic solvents, chaotropic anions). These results indicated that the modification of hydrophobic (intramolecular) interactions in chloroplast fructose-1,6-bisphosphatase constituted the underlying mechanism in light-activation by the ferredoxin-thioredoxin system. PMID- 2555191 TI - Inhibition of Desulfovibrio gigas hydrogenase with copper salts and other metal ions. AB - The effect of several transition metals on the activity of Desulfovibrio gigas hydrogenase has been studied. Co(II) and Ni(II) at a concentration of 1 mM did not modify the activity of the enzyme; nor did they affect the pattern of activation/deactivation. Cu(II) inhibited the active hydrogenase, prepared by treatment with hydrogen, but had little effect on the 'unready' enzyme unless a reductant such as ascorbate was present, in which case inactivation took place either in air or under argon. Hg(II) also inactivated the enzyme irreversible in the 'unready' state without the requirement for reductants. The reaction of H2 uptake with methyl viologen was much more sensitive to inhibition than the H2/tritium exchange activity. EPR spectra of this preparation showed that the rates of decline were [3Fe-4S] signal greater than H2-uptake activity greater than Ni-A signal. Similar results were obtained when the protein was treated with Hg(II). The results demonstrate that the [3Fe-4S] cluster is not essential for H2 uptake activity with methyl viologen, but the integrity of [4Fe-4S] clusters is probably necessary to catalyze the reduction of methyl viologen with hydrogen. D. gigas hydrogenase was found to be highly resistant to digestion by proteases. PMID- 2555192 TI - Phosphorylation of low molecular mass cytosolic proteins by protein kinase C and protein kinase A in the rabbit exocrine pancreas. AB - Subcellular fractionation of rabbit pancreatic acini was performed to study the distribution of endogenous substrates for protein kinase C. Substrates for protein kinase C were found to be predominantly low molecular mass proteins of cytosolic origin. At least three of these soluble substrates, with molecular masses of 17-19 kDa, were relatively heavily phosphorylated by endogenous as well as purified pancreatic protein kinase C. In the same molecular mass range, 16-18 kDa, soluble proteins were also phosphorylated by protein kinase A. Moreover, addition of cyclic AMP under conditions that activated protein kinase C gave a more than additive labelling of these low molecular mass proteins. The latter observation may be of interest in view of the potentiating effect cyclic-AMP activated protein kinase A has on amylase secretion stimulated by secretagogues which increase free cytosolic Ca2+ and activate protein kinase C. PMID- 2555193 TI - Degradation of ornithine decarboxylase in mammalian cells is ATP dependent but ubiquitin independent. AB - Ornithine decarboxylase (ODC), a key enzyme in the biosynthesis of polyamines in mammalian cells is characterized by an extremely short half-life. In the present study, ODC degradation was investigated in 653-1 mouse myeloma cells that overproduce ODC and in ts85 cells that are thermosensitive for conjunction of ubiquitin to target proteins. Addition of 2-deoxyglucose and dinitrophenol (agents that efficiently deplete cellular ATP) to the growth medium of these cells inhibited ODC degradation. In contrast, chloroquine and leupeptin, inhibitors of intralysosomal proteolysis, did not affect ODC degradation. Shifting ts85 cells to 42 degrees C (a non-permissive temperature that inhibited conjugation of ubiquitin to target proteins) did not prevent ODC degradation. The ATP-dependent degradation of ODC in 653-1 cells was inhibited substantially by N alpha-tosyl-L-lysine chloromethane (TosPheMeCl), iodoacetamide and o phenanthroline. These results suggest that ODC degradation occurs via a non lysosomal. ATP-requiring and ubiquitin-independent cellular proteolytic mechanism, and that serine proteases and enzymes containing sulphydryl groups and metalloenzyme(s) may be involved in this process. PMID- 2555194 TI - Characterization of a membrane-associated phosphotyrosyl protein phosphatase from the A431 human epidermoid carcinoma cell line. AB - The A431 human epidermoid carcinoma cell line exhibits a 30-100-fold overexpression of the epidermal growth factor (EGF) receptor. We have characterized a membrane-associated phosphotyrosyl-protein phosphatase (PTPase) in these cells since it seemed reasonable that overexpression of the EGF-receptor tyrosine kinase will be matched by high PTPase activity. Indeed, of 12 cell lines tested, the A431 cells had the highest specific PTPase activity. About 70% of the total cellular PTPase activity was found associated with membranes after cell fractionation. The membrane-associated PTPase was hydrophobic as judged by its behaviour in Triton X-114 phase partitioning. High-performance liquid chromatography (HPLC) on a DEAE column revealed a single, homogeneous species of membrane-associated PTPase with an apparent molecular mass of 43 kDa as determined by HPLC on a gel permeation column in the presence of Triton X-100. Comparison of this PTPase with the membrane-associated PTPase activities present in rat spleen and in the human chronic myelogenous leukemia cell line K562 revealed additional species resolvable by DEAE-HPLC. These findings suggest that cells may possess different PTPase activities depending on their growth and differentiation states. PMID- 2555195 TI - Extracellular but not intracellular calcium mobilization is required for Epstein Barr virus-containing supernatant-induced B cell activation. AB - Changes in intracytosolic free calcium concentration ([Ca2+]i) are though to be an important trigger for the initiation of the cellular events culminating in B cell activation. After exposure of human B lymphocytes loaded with the fluorescent indicator quin-2 to the Epstein-Barr virus (EBV)-containing supernatant of B95-8 cell line, a rise in [Ca2+]i is observed. To determine the respective contribution of the intra- and extracellular Ca2+ pools in EBV-induced B cell activation, the pharmacologic modulation of these processes was investigated using an intracellular calcium chelator, 1,2-bis(o aminophenoxy)ethane- N,N,N',N'-tetraacetic (BAPTA) or the calcium channel blocking drugs. When the extracellular Ca2+ contribution was minimized in the presence of calcium channel-blocking drugs or incubation of the cells in Ca2+ free medium, the EBV-induced human B cell activation was fully inhibited. When the calcium channel-blocking drugs, either verapamil or diltiazem, were withdrawn or when exogenous Ca2+ was added to the Ca2+-free medium, EBV-induced B cell activation was noted, demonstrating the reversibility of the inhibition. On the contrary, when the intracellular Ca2+ contribution was reduced after loading the cell with BAPTA, no alteration of the EBV-induced B cell activation was observed. Thus, the EBV-induced rise of [Ca2+]i required in the activation of human B cells appears to be essentially related to the entry of extracellular Ca2+ and not to the release of Ca2+ from internal stores. PMID- 2555196 TI - Regulation of surface IgM- and IgD-mediated inositol phosphate formation and Ca2+ mobilization in murine B lymphocytes. AB - Cross-linking of surface IgM or IgD receptors on B cells initiates a signaling cascade involving the activation of an (uncharacterized) G-protein: this in turn activates a polyphosphoinositide-specific phosphodiesterase (PPI-PDE), thereby leading to the release of inositol phosphates. In order to investigate if the two isotypes of sIg share a common G-protein, we stimulated B cells sequentially with anti-mu and anti-delta antibodies. Ligation of either class of receptor for 1 h led to the activation of the PPI-PDE, which persisted for several hours. However, this was accompanied by inhibition of further stimulation of the enzyme via the heterologous receptors. This desensitization was shown to operate at the level of the coupling between G-protein and the PPI-PDE. These effects waned after 4-8 h of stimulation, when signaling via the heterologous receptors had essentially returned to normal. In addition, stimulation of B cells by anti-mu and anti-delta together did not elicit additive responses, either in terms of increases in inositol phosphate production, or in terms of increases in intracellular Ca2+ levels. Taken together, these results indicate that sIgM and IgD receptors share a common G-protein and that signaling via these receptors is under both positive and negative feedback control. The mechanisms involved are unknown, but these effects may well be due to modulation of the activities of components of the signaling cascade by protein kinase C. PMID- 2555197 TI - cAMP inhibits the OKT3-induced increase in cytoplasmic free calcium in the Jurkat T cell line: the degree of inhibition correlates inversely with the amount of CD3 binding ligand used. AB - We have investigated the effect of cAMP concentration on the CD3-mediated rise in intracellular Ca2+ induced by anti-CD3 monoclonal antibody in the human T cell leukemia line Jurkat. Forskolin, prostaglandin E2 (PGE2) and dibutyryl cAMP (db cAMP) were used to increase intracellular cAMP concentrations. Treatment of Jurkat cells with forskolin or db-cAMP for 3 h inhibited the subsequent rise in intracellular Ca2+ concentration induced by an optimally mitogenic dose of 100 ng/ml of the anti-CD3 OKT3, whereas PGE2 counteracted the Ca2+ rise only marginally. The inhibitory effect of forskolin and PGE2 on the Ca2+ signal correlated with their abilities to induce increased cAMP levels in Jurkat cells. The suppression of the Ca2+ response was dependent on the concentration of the cAMP-elevating agent and the time of pre-incubation with the drug. The cAMP mediated inhibition of the Ca2+ response diminished or disappeared when increasing concentrations of OKT3 were used to stimulate the rise in intracellular Ca2+ concentration. The results indicate that cAMP participates in the regulation of T lymphocyte activation at the level of signal transduction. Our findings, which stress the crucial role of the concentration of the ligand used to trigger Ca2+ responses, provide an explanation to previous contradictory reports on the impact of cAMP on the signal transduction in T cells. PMID- 2555198 TI - Increased peripheral alpha 1-adrenoceptor sensitivity following chronic thioridazine treatment in the pithed rat. AB - The effect of chronic treatment (10 days) with thioridazine (25 mg/kg per day, i.p.) on the increase in diastolic blood pressure induced by methoxamine and xylazine was studied in the pithed rat. The dose-response curve for methoxamine, but not for xylazine, was shifted to the left by the neuroleptic. These results indicate that postjunctional alpha 1-adrenoceptors in the rat blood vessels become supersensitive after long-term treatment with thioridazine. PMID- 2555199 TI - Insensitivity of the open state of (-)-DPI-modified cardiac Na+ channels to the blocking effects of the (+) enantiomer. AB - Elementary Na+ currents in DPI-modified cardiac Na+ channels were recorded in the absence and presence of the blocking (+) enantiomer in inside-out patches from cultured neonatal rat cardiocytes. After external application of (-)-DPI (3 mumols/l), the kinetics of modified, non-inactivating Na+ channels were found to be indistinguishable from the channel kinetics observed in the presence of (+/-) DPI (6 mumols/l). The open state and burst duration proved insensitive to the blocking (+) enantiomer in concentrations up to 3 mumols/l. At -30 mV, the exit rates from the open and the bursting state were 105 +/- 11 and 57 +/- 6 s-1, respectively, with (-)-DPI or 88 +/- 6 and 49 +/- 8 s-1, respectively, with (+/-) DPI. The use of (+/-)-DPI 201-106 as a pharmacological modifier of channels will, therefore, provide reliable kinetic data of non-inactivating Na+ channels. PMID- 2555200 TI - A progesterone metabolite enhances the activity of the GABAA receptor complex at the pituitary level. AB - The interaction of 5 alpha-pregnane-3 alpha-ol-20-one (5 alpha 3 alpha P), a progesterone metabolite, with the GABAA receptor chloride channel complex was investigated at the pituitary level. In nanomolar concentrations this steroid potentiated the inhibitory effect of muscimol (a GABAA agonist) on prolactin release from rat pituitary cells in culture. In micromolar concentrations 5 alpha 3 alpha P had a direct inhibitory effect, similar to that of muscimol, with an IC50 value of 370 nM. This effect was antagonized by bicuculline, a GABAA antagonist, and by picrotoxin, a chloride ion channel blocker. Its reduced isomer, 5 alpha 3 beta P, and progesterone (Pg) were devoid of activity. Using [35S]t-butylbicyclophosphorothionate ([35S]TBPS) as a ligand, we demonstrated, for the first time, specific barbiturate sites on pituitary membranes, similar to those of the central nervous system, with a Kd value of 25 nM and a Bmax value of 62 fmol/mg protein. 5 alpha 3 alpha P inhibited the binding of [35S]TBPS. In contrast, its 3 beta isomer was inactive. These data show that 5 alpha 3 alpha P enhanced the activity of the GABAA receptor complex at the pituitary level and suggest that its inhibitory effect on prolactin release might be mediated by the barbiturate site or by a closely related site. PMID- 2555201 TI - Platelet-activating factor causes goblet cell depletion in the conjunctiva. AB - Platelet-activating factor (PAF) (1-O-hexadecyl-2-acetyl-sn-glyceryl-3 phosphorylcholine) produced dose-dependent depletion of the goblet cell population associated with the conjunctival epithelium. Reductions in goblet cell numbers did not correspond to leukocyte infiltration and were consistent with a direct effect of PAF. In contrast, LTD4 and LTE4 did not affect the goblet cell population although they caused massive eosinophil infiltration into the conjunctival epithelium. Histamine also produced conjunctival goblet cell depletion, but this appeared secondary to eosinophil degranulation and resultant epithelial desquamation. In addition to goblet cell expulsion, PAF produced an increase in conjunctival microvascular permeability over an identical dose-range. PAF-induced leukocyte emigration was small or absent and comprised a neutrophil infiltrate which exhibited no clear dose-dependent relationship. Lyso-PAF produced effects only at the highest dose employed where pathological changes and a distinct increase in conjunctival microvascular permeability were evident. Lyso PAF- and PAF-induced increases in conjunctival microvascular permeability were virtually abolished by the PAF antagonist CV-6209. The pronounced inhibitory activity of CV-6209 suggests that high doses of lyso-PAF may either weakly stimulate conjunctival PAF receptors or that there may be sufficient conversion of lyso-PAF to biologically active levels of PAF. PMID- 2555202 TI - Diacylglycerol in the synergy of bradykinin and thrombin stimulation of prostaglandin synthesis. AB - Simultaneous addition of bradykinin and thrombin to 3T3 fibroblasts for 5 min resulted in less than additive stimulation of prostaglandin E2 synthesis. However, if cells were stimulated with either agonist alone, then the other added 15 min later, prostaglandin E2 synthesis was synergistically enhanced. In contrast, if either agonist was added, then prostaglandin E2 synthesis in response to the same agonist assessed 15 min later, synthesis was markedly reduced. Bradykinin and thrombin caused increased diacylglycerol accumulation in the cells, and addition of the diacylglycerol kinase inhibitor R59022 dramatically increased the effects of sequential addition of the agonists. These results suggest that diacylglycerol generated in response to activation of one receptor amplifies the effects of activation of other receptors. PMID- 2555203 TI - The effect of endothelin on noradrenergic transmission in rat and guinea-pig atria. AB - Pre-and postjunctional effects of 10 nM endothelin have been studied in rat and guinea-pig spontaneously beating isolated atria. Endothelin increased the rate and force of contraction of atria from both species, but decreased the positive inotropic and chronotropic responses to stimulation of intramural sympathetic nerves by approximately 60% in rat and by 10-15% in guinea-pig atria. The stimulation-induced release of noradrenaline from either rat or guinea-pig atria was not significantly changed by endothelin. PMID- 2555204 TI - Suppression of calcium current in a snail neurone by eperisone and its analogues. AB - The effects of eperisone, tolperisone and isoperisone on the calcium current (ICa) were studied in an identified neurone of Achatina fulica under voltage clamping. At a holding voltage of -50 mV, these drugs inhibited ICa dose dependently without affecting activation time. Eperisone (IC50 = 0.348 mM) and isoperisone (0.226 mM) were significantly more effective than tolperisone (1.089 mM). Eperisone binds competitively with Ca2+ to the Ca2+ channels, based on its effects seen with various extracellular Ca2+ concentrations. The three drugs shifted the steady state inactivation curves in the hyperpolarizing direction. The mean dissociation constants for inactivated Ca2+ channels were calculated to be 0.070 mM (eperisone), 0.162 mM (tolperisone) and 0.014 mM (isoperisone). These values were much lower than their IC50 at Vh of -50 mV, which are reflected as the dissociation constants for resting Ca2+ channels. High frequency stimulation did not potentiate ICa suppression, suggesting that the drugs hardly bind to activated Ca2+ channels. These findings indicate that the selectivities of the Ca2+ channels to the drugs are in the order of their inactivated, resting and open states. PMID- 2555205 TI - Different mu receptor subtypes mediate spinal and supraspinal analgesia in mice. AB - To examine the relative roles of mu 1- and mu 2-receptors in spinal and supraspinal analgesia, we assessed the effects of naloxonazine, naloxone, beta funaltrexamine (beta-FNA), and ICI-154,129 on tail-flick analgesia produced by intrathecal or intracerebroventricular injections of the highly mu-selective agonist, [D-Ala2,Me-Phe4,Gly(ol)5]enkephalin (DAGO; mu 1 and mu 2), [D Ser2,Leu5]enkephalin-Thr6 (DSLET; mu 1 and delta), and the selective delta receptor agonist [D-Pen2,D-Pen5]enkephalin (DPDPE) in mice. Both DAGO and DSLET supraspinal analgesia were mediated through mu 1-receptors. Naloxonazine shifted the supraspinal DAGO dose-response curve 4-fold to the right without changing the curve for spinal DAGO. Likewise, naloxonazine pretreatment shifted supraspinal DSLET analgesia 10-fold, whereas spinal DSLET analgesia was not affected. DPDPE analgesia was not antagonized spinally or supraspinally by naloxonazine pretreatment. These findings suggest that DAGO produces analgesia spinally and supraspinally through different sets of mu-receptors. Moreover, at least two distinct receptor subtypes mediated spinal analgesia. First, naloxone inhibited spinal DAGO analgesia more potently than DPDPE analgesia. Second, the irreversible mu-antagonist, beta-FNA, blocks spinal DAGO analgesia. Since spinal DAGO was insensitive to naloxonazine, ruling out a mu 1 mechanism, these results indicate a role for mu 2-receptors. Spinal DAGO analgesia also developed tolerance to morphine far more slowly than supraspinal DAGO analgesia even though mu-receptors mediate both, as indicated by their sensitivity towards beta-FNA. Finally, the delta-antagonist ICI-154,129 is a more potent inhibitor of spinal DPDPE analgesia than spinal DAGO analgesia. Thus, delta-receptors mediate spinal DPDPE analgesia. PMID- 2555206 TI - Stereoselective binding of niguldipine enantiomers to alpha 1A-adrenoceptors labeled with [3H]5-methyl-urapidil. AB - [3H]5-Methyl-urapidil, a potent antihypertensive derivative of urapidil, binds to alpha 1A-adrenoceptors in rat brain cortex membranes with a dissociation constant (KD) of 0.89 nM and a Bmax of 116 fmol/mg protein. The ligand does not bind to purified liver cell membranes (alpha 1B-adrenoceptors). [3H]5-Methyl-urapidil also labels 5-HT1A receptors in brain membranes (KD: 0.84 nM and Bmax: 235 fmol/mg protein). (+/-)-Niguldipine, a novel 1,4-dihydropyridine with Ca2+ antagonistic as well as alpha 1A-adrenoceptor blocking properties, is a competitive inhibitor of [3H]5-methyl-urapidil binding to alpha 1A-adrenoceptors. In contrast to those for prazosin, the Ki values for niguldipine were highly dependent on the membrane protein concentration, indicating partitioning of niguldipine into hydrophobic compartments unavailable for alpha-adrenoceptor interaction. The extrapolated, 'true' Ki values were as follows: (+/-) niguldipine: 0.298 nM, (-)-niguldipine: 3.12 nM, (+)-niguldipine: 0.145 nM. PMID- 2555207 TI - Reconstitution of the solubilized mu-opioid receptor coupled to a GTP-binding protein. AB - A mu-opioid receptor-GTP binding protein (mu-opioid receptor-G-protein) complex from the 7315c cell was solubilized with CHAPS (3-[(3 cholamidopropyl)dimethylammonio]-1-propane sulfonate) and reconstituted into phospholipid vesicles. Pretreatment of the tissue with either [3H]etorphine or morphine greatly improved recovery of the receptor and maintained it in a GTP sensitive state. GTP sensitivity was consistent with the hypothesis that a receptor-G-protein complex had been obtained. Other evidence consistent with this hypothesis was that recovery of the solubilized, prelabelled receptor was decreased by approximately 70% by pretreatment of 7315c cells with pertussis toxin. The reconstituted receptor was mu-selective: DAGO (Tyr-D-Ala-Gly-Met-Phe- NH(CH2)2OH), but not ICI 174864 or U50488-H, displaced [3H]etorphine binding with high affinity. The affinity of the reconstituted receptor for [3H]etorphine (1.25 +/- 0.20 nM) was similar to that observed for the membrane-associated receptor (0.53 +/- 0.25 nM). GTP gamma S decreased this affinity 3-fold without changing the number of binding sites. The potencies of GTP gamma S and GTP in diminishing [3H]etorphine binding were similar in the membrane and vesicle preparations, but were 10-fold lower than the potencies observed in diminishing binding to the solubilized receptor. The ability to reconstitute a functional mu-opioid receptor G-protein complex will facilitate further study of the structure and function of the receptor and the specific identification of the associated GTP-binding protein(s). PMID- 2555208 TI - Effect of aging on properties and function of beta-adrenoceptors in rat lung. AB - beta-Adrenoceptor density and ligand affinity, basal adenylate cyclase activity and cAMP synthesis upon stimulation with forskolin, fluoride, guanine nucleotides (GTP or guanylyl imidodiphosphate (GppNHp) or isoproterenol in the presence of the nucleotides were studied in membranes prepared from lungs of young (aged 2-3 months) and of old (aged 24-25 months) male Wistar rats. There was a significant (P less than 0.05, 21%) increase in beta-receptor density and a significant (P less than 0.05, 38%) decrease in the percentage of high-affinity binding sites for isoproterenol. Both basal adenylate cyclase activity and that after stimulation with guanine nucleotides or isoproterenol in the presence of nucleotides were unaltered with age. Forskolin stimulation of cAMP synthesis was significantly reduced (by 24%, P less than 0.05) in tissues from older animals. It is suggested that the age-dependent changes in properties of beta-receptors in rat lungs are compensatory, in order to ensure equal cAMP production for equal agonist stimulation. PMID- 2555209 TI - Forskolin inhibition of cyclic AMP generation in J774 macrophages. AB - Forskolin inhibited cyclic AMP generation in J774 macrophage cells in response to isoproterenol. Forskolin, 10 nM-0.1 mM, also inhibited the adenylate cyclase activity of membrane preparations. The basal activity and the isoproterenol-, cholera toxin-, fluoride- or GppNHp-stimulated activities were maximally depressed by 10 microM forskolin (30-70% inhibition, EC50 = 0.3-0.5 microM). This effect was achieved similarly in membranes from pertussis toxin-treated cells. Forskolin required guanine nucleotides for inhibition. In the absence of GTP the decrease in basal activity was reversed into stimulation (EC50 = 10 microM forskolin). Reversal of inhibition into activation also followed the addition of 1 mM MnCl2 (EC50 = 10 microM forskolin). 1,9-Dideoxyforskolin was ineffective to alter adenylate cyclase activity. In contrast, a water-soluble derivative of forskolin was as active as forskolin to regulate activity. The results suggest that forskolin may interact with adenylate cyclase to cause either activation or inhibition depending on the degree of activation of Ns and on its interaction with the catalyst. PMID- 2555210 TI - Human HT-29 colon carcinoma cells contain muscarinic M3 receptors coupled to phosphoinositide metabolism. AB - Five different muscarinic receptor subtypes can be distinguished by the differences in their amino acid sequence, the coupled signal transduction system, pharmacological binding properties and activation of ionic fluxes. The present study served to characterize the binding profile of muscarinic receptors in human colon carcinoma cells (HT-29) using selective muscarinic antagonists. The affinities of the compounds were compared with their potency to inhibit cholinergically-activated phosphoinositide metabolism. Pirenzepine displaced [3H]N-methyl-scopolamine binding and inhibited inositolphosphate (IP) release with potencies typical of those of non-M1 receptors. The M3 subtype-selective antagonists sila-hexocyclium and hexahydro-sila-difenidol had high affinity to the muscarinic receptors in HT-29 cells (KD = 3.1 nM and 27 nM, respectively) and inhibited IP release at nanomolar concentrations. The M2 receptor antagonists, AF DX 116 and methoctramine, had low antimuscarinic potencies. Our results demonstrate that HT-29 human colon carcinoma cells contain an apparently pure population of M3 receptors. These cells could serve as a model system for further investigations concerning regulatory and signal transduction mechanisms associated with glandular muscarinic M3 receptors. PMID- 2555211 TI - NMDA receptor activation induces nitric oxide synthesis from arginine in rat brain slices. AB - Activation of N-methyl-D-aspartate (NMDA) receptors in rat cerebellum leads to the release of endothelium-derived relaxing factor, now identified as nitric oxide (NO), a stimulator of soluble guanylate cyclase. L-NG-monomethylarginine (L NMMA), which blocks NO synthesis from L-arginine in several tissues, including a crude synaptosomal preparation from brain, inhibited the elevation of cyclic GMP induced by NMDA in rat cerebellar slices. D-NMMA was ineffective. L-Arginine, but not its D enantiomer, augmented the response to NMDA and reversed the inhibition by L-NMMA. The results indicate that stimulation of NMDA receptors results in the activation of the enzyme which catalyzes the formation of NO from L-arginine. PMID- 2555212 TI - Hemin-induced erythroid differentiation changes the sensitivity of K562 cells to tumor necrosis factor-alpha. AB - Tumor necrosis factor-alpha (TNF) can inhibit the growth of erythroid progenitors (erythroid colony-forming units [CFU-E] and erythroid burst-forming units [BFU E]) at picomolar concentrations, but only if added within the first 48 h of culture. These data suggested that cells undergoing erythroid differentiation become resistant to TNF. To test this hypothesis, K562 cells were treated with hemin to induce erythroid differentiation and then tested for their sensitivity to TNF in terms of growth and TNF receptor expression. TNF inhibited the growth of untreated K562 cells, but not hemin-treated K562 cells. Untreated K562 cells expressed TNF receptors, whereas few hemin-treated K562 cells expressed TNF receptors within 24 h of exposure to hemin. These data show that K562 cells induced to differentiate along the erythroid pathway are resistant to TNF because they lack TNF receptors and suggest that the resistance of erythropoietin-treated human bone marrow cells to TNF added after 48 h of culture may also reflect loss of TNF receptors associated with erythroid differentiation. PMID- 2555213 TI - Effects of recombinant tumor necrosis factor (rHuTNF alpha) on human neutrophils and monocytes: in vitro, ex vivo and in vivo. AB - In in vitro experiments, tumor necrosis factor (rHuTNF alpha) was found to inhibit spontaneous and directed migration of normal human neutrophils and monocytes. RHuTNF alpha showed no chemoattractant activity. In conjunction with a Phase-2 clinical trial, we also studied in vivo rHuTNF alpha effects on neutrophil and monocyte number and function. At the time of maximal plasma TNF levels (30 min), marked neutropenia and monocytopenia were observed. Isolated neutrophils were activated for superoxide production but were unable to migrate. Monocyte migration was inhibited at later times. Neutrophil migratory function recovered between 4 and 8 h but was again depressed at 24 h. The data demonstrate the complexity of the response to TNF, comprising direct and indirect effects which are concentration-, time- and place-dependent. They further suggest that the only neutrophils and monocytes available for participation in an anti-tumor activity of TNF in vivo are those present in the tumor at the outset. PMID- 2555214 TI - Discordance in the development of peripheral and autonomic neuropathy during vincristine therapy. PMID- 2555216 TI - Two different forms of endothelin receptors in rat lung. AB - We provided evidence for the presence of two distinct types of rat endothelin (ET) receptors having different molecular masses by affinity labelling of rat lung membranes with 125I-ET-1, ET-2 and ET-3 and SDS-PAGE followed by autoradiography. One type, which is 44 kDa has a higher affinity for ET-1 and -2 than ET-3, whereas the other type, which is 32 kDa, preferably interacts with ET 3 rather than ET-1 and -2. PMID- 2555215 TI - Testing cell activation and cytokine release in serum-supplemented cultures: methodical note concerning polymyxin B. PMID- 2555217 TI - Thyroid hormone stimulates expression of 6-phosphofructo-2-kinase in rat liver. AB - The activity of liver 6-phosphofructo-2-kinase (PFK-2), the enzyme that catalyses the synthesis of fructose 2,6-bisphosphate, was markedly decreased in hypothyroid rats and partially restored after 3 days of treatment with triiodothyronine. The changes in PFK-2 activity were accompanied by parallel changes in enzyme content measured by immunotitration and in PFK-2 mRNA determined by dot blot and Northern blot hybridization with cDNA probes. It is concluded that thyroid hormone stimulates liver PFK-2 gene expression by a pre-translational mechanism. PMID- 2555218 TI - Tyrphostins inhibit the epidermal growth factor receptor-mediated breakdown of phosphoinositides. AB - In response to epidermal growth factor (EGF) and the Ca2+ ionophore A23187, the total phosphatidylinositides (IPT) increased in A431 human epidermoid carcinoma cells 1.8- and 2.0-fold and in the EGF-dependent A431/Clone 15-2 cells 3.0- and 8.0-fold, respectively, over basal levels. Both responses were inhibited by the antiproliferative agents tyrphostins, but the EGF-induced increase in IPT was inhibited to a much greater extent than that induced by the ionophore. Tyrphostins which are potent EGF-receptor kinase inhibitors were also potent in blocking the EGF-induced production of phosphoinositides. The less potent tyrphostins were found to inhibit the EGF-dependent IPT formation more weakly. These results support the notion that phospholipase C is activated through its phosphorylation by the EGF receptor. PMID- 2555219 TI - GPC phosphodiesterase and phosphomonoesterase activities of renal cortex and medulla of control, antidiuresis and diuresis rats. AB - Glycerylphosphorylcholine (GPC) concentration was reported to be elevated in renal medulla of experimental animals deprived of water. The activities of GPC phosphodiesterases were similar in homogenates and membrane subfractions of renal cortex prepared from control, diuresis and antidiuresis rats. There were no differences in these preparations' ability to hydrolyze phosphorylcholine. In contrast, there was a nearly 50% reduction of non-specific phosphomonoesterase activity, using p-nitrophenylphosphate as substrate and membrane subfractions prepared from the antidiuresis animals. It is suggested that as a consequence, a pathway for the formation from L-alpha-glycerylphosphate is activated. PMID- 2555220 TI - A type 2A protein phosphatase dephosphorylates the elongation factor 2 and is stimulated by the phorbol ester TPA in mouse epidermis in vivo. AB - Mouse epidermal cytosol contains a protein phosphatase with Mr 38,000, which dephosphorylates the elongation factor 2 (EF-2) of protein biosynthesis and is stimulated after topical application of TPA to mouse skin [(1988) Biochem. Biophys. Res. Commun. 153, 1129-1135]. Dephosphorylation of EF-2 by this phosphatase is inhibited by okadaic acid at concentrations as low as 10(-8) M, but not by heparin up to concentrations of 600.micrograms/ml. The catalytic subunit of protein phosphatase 2A (PP2Ac) with EF-2 as a substrate exhibits the same sensitivity towards okadaic acid and insensitivity towards heparin as the EF 2 phosphatase of epidermal cytosol. The catalytic subunit of protein phosphatase 1 (PP1c) is strongly suppressed by heparin and less sensitive towards okadaic acid than PP2Ac. PP2Ac is around 50 times more efficient in dephosphorylating EF 2 than PP1c. These data indicate that the TPA-stimulated EF-2 phosphatase in epidermal cytosol is a type 2A protein phosphatase. PMID- 2555221 TI - The dependence on intracellular ATP concentration of ATP-sensitive K-channels and of Na,K-ATPase in intact HIT-T15 beta-cells. AB - We have studied the effects of changes of intracellular ATP concentration ([ATP]i) on the activity of ATP-sensitive K-channels (IK(ATP] and of Na,K-ATPase in intact cells of the insulin-secreting cell-line HIT-T15. Pre-exposure of HIT beta-cells to oligomycin caused a dose-dependent reduction in [ATP]i. Marked activation of IK(ATP) activity was found when ATP was lowered below 3 mM. Na,K ATPase was progressively inhibited as ATP was lowered to 1.5 mM. These data demonstrate that changes in intracellular ATP in the millimolar range markedly influence the activity of two beta-cell membrane proteins having affinities for ATP in the micromolar range. This suggests that submembrane [ATP] may be considerably below the measured bulk cytosolic concentration. The findings also support the proposed role of intracellular ATP in mediating effects of changes in glucose concentration on the activity of beta-cell IK(ATP) and insulin secretion. PMID- 2555222 TI - A theoretical study of the acidification of the rhinovirus capsid. AB - Electrostatic calculations for human rhinovirus 14 indicate that histidine-base residue pairs in the region of a beta-strand interaction between pentamers may be involved in a pH-induced process that leads to the release of viral RNA. Other picornavirus sequences are examined for these residue pairs, a subset of which is present in enteroviruses. Foot and mouth disease virus possesses one of the residue pairs, and cardioviruses, which undergo a separate pH and halide ion induced capsid dissociation, possess none. PMID- 2555223 TI - Phosphorylation of caldesmon. AB - The phosphorylation of caldesmon was studied to determine if kinase activity reflected either an endogenous kinase or caldesmon itself. Titration of kinase activity with calmodulin yielded maximum activity at substoichiometric ratios of calmodulin/caldesmon. The sites of phosphorylation on caldesmon for calcium/calmodulin-dependent protein kinase II and endogenous kinase were the same, but distinct from protein kinase C sites. Phosphorylation in the presence of Ca2+ and calmodulin resulted in a subsequent increase of endogenous kinase activity in the absence of Ca2+. These results suggest that caldesmon is not a protein kinase and that kinase activity in caldesmon preparations is due to calcium/calmodulin-dependent protein kinase II. PMID- 2555224 TI - Activation of protein kinase C alters the interaction of alpha 2-adrenoceptors and the inhibitory GTP-binding protein (Gi) in human platelets. AB - The effect of 12-tetradecanoyl phorbol 13-acetate (TPA) on the hormonal modulation of adenylate cyclase was studied. The effect of epinephrine (alpha 2 adrenergic action) was markedly diminished in membranes from TPA-treated platelets as compared to the controls. Interestingly, the inhibitory effect of guanylyl imido diphosphate (Gpp(NH)p) was not altered. Neither the number of alpha 2-adrenoceptors nor their affinity for [3H]yohimbine were affected by the treatment with TPA. In control platelets, 77% of the receptors were in a high affinity state for epinephrine and 22% in a low-affinity state; Gpp(NH)p shifted the receptor affinity towards the low-affinity conformation. In membranes from TPA-treated platelets, the receptors were in the low-affinity state and no further decrease in affinity was induced by Gpp(NH)p. Our data suggest that activation of protein kinase C in platelets blocks the hormonal inhibition of adenylate cyclase by interfering with the receptor-Gi interaction. PMID- 2555225 TI - Human Na+,K+-ATPase genes. Beta-subunit gene family contains at least one gene and one pseudogene. AB - The existence of a chromosome gene family containing at least one gene and one pseudogene was shown for the Na+,K+-ATPase beta-subunit. A partial structure of the beta 1-gene was determined, the coding part of which was completely homologous to cDNA of the Na+,K+-ATPase beta I-subunit from HeLa cells. The region encoding the putative protein transmembrane domain was shown to be bordered by two introns. The structure of a pseudogene (beta psi) was determined. This pseudogene is processed and contains multiple stop codons. Its homology to the beta I-subunit cDNA from HeLa cells is about 88%. PMID- 2555226 TI - [Adrenergic receptors regulation of insulin secretion due to 12-O-tetradecanoyl phorbol-13-acetate--a perifusion study]. AB - A possible role for the activation of protein kinase C and adenylate cyclase in the regulation of insulin secretion by monolayer islet B cells of neonatal rats was examined in a perifusion system. Stimulation of 60 min of 16.7 mM glucose secreted a first component of insulin for the first 10-min period. In the presence of 200 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA), a protein kinase C activator, however, the same dose of glucose significantly increased cAMP accumulation in medium and dramatically enhanced a second component of insulin secretion in which the average secretion rate was 1.81 +/- 0.25 of insulin for the first 10-min period. In the presence of 200 nM 12-O-tetradecanoyl phorbol-13 acetate(TPA), a protein kinase C activator, however, the same dose of glucose significantly increased cAMP accumulation in medium and dramatically enhanced a second component of insulin secretion in which the average secretion rate was 1.81 +/- 0.25 of insulin for the first 10-min period. In the presence of 200 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA), a protein kinase C activator, however, the same dose of glucose significantly increased cAMP accumulation in medium and dramatically enhanced a second component of insulin secretion in which the average secretion rate was 1.81 +/- 0.25 ng/ml/min for the 10-30 min (second phase IIa) and 3.39 +/- 0.18 ng/ml/min for the 30-60 min (second phase IIb) respectively. Addition of 10 microM epinephrine significantly decreased the medium cAMP content and abolished the sustained second-phase secretion, resulting in a monophasic response of approximately the same magnitude as evoked by 16.7 mM glucose alone. In contrast, joint addition of 10 microM epinephrine and 10 microM phentolamine, which activates adenylate cyclase, exerted a more pronounced increase in cAMP accumulation and the second component of insulin secretion than in the presence of TPA; the average secretion rates were 3.62 +/- 0.38 and 6.44 +/- 0.35 ng/ml/min respectively for the second phase IIa and IIb. However, no antagonistic effect of propranolol (10 microM) was found. Likewise, either phenylephrine (10 microM) or clonidine (1 microM) inhibited enhancement of insulin secretion that a combination of glucose with TPA caused, and alpha 1 adrenergic blockade of 10 microM prazosin was weak as compared to the blockade of yohimbine at the same concentration.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2555227 TI - Effect of novobiocin on gamma-radiation-induced DNA repair in human lymphocyte subpopulations. AB - 1. DNA repair was measured in 3 Gy gamma-irradiated human peripheral lymphocyte subpopulations by means of nucleoid sedimentation. 2. The influence of the antibiotic, novobiocin (an inhibitor of inter alia topoisomerase II) on the repair process was investigated. 3. Repair of 33 37% of the DNA lesions induced by gamma-irradiation in enriched B lymphocyte fractions, was retarded by novobiocin. 4. Repair in enriched T lymphocyte fractions was unaffected by novobiocin. PMID- 2555228 TI - The role of autophosphorylation of cAMP-dependent protein kinase II in the inhibition of protein phosphatase-1. AB - 1. The inhibition of the catalytic subunit of protein phosphatase-1 (PP-1c) by the regulatory subunit of cAMP-dependent protein kinase II (RII) was studied. 2. Phosphorylation or thiophosphorylation of RII increased its inhibitory potency up to 4- and 6-fold and rendered it competitive with respect to the substrate of PP 1c, phosphorylase a. The Ki values for thiophospho-RII and phospho-RII were 200 and 500 nM, respectively. 3. Though PP-1c was able to release phosphate from phospho-RII, its activity once incubated with phospho-RII, remained inhibited even 80% of the phosphate was released from phospho-RII. 4. The catalytic subunit of cAMP-dependent protein kinase was effective in suspending the inhibition employed either before or after the addition of phospho-RII to PP-1c. 5. No exclusive bindings of thiophospho-RII and heat-stable protein inhibitors to the PP-1c could be proved by double inhibition studies, however some synergism was observed in their effect. PMID- 2555229 TI - Effect of phospholipase C treatment on the activity of the particulate cyclic nucleotide phosphodiesterase of rat brain. AB - 1. The activity of cAMP phosphodiesterase (PDE) was studied in a 10,000 g particulate fraction prepared from rat brain. 2. Phospholipase C such as sphingomyelin choline phosphodiesterase (SMase), phosphatidylinositol phosphodiesterase (PIase) and phosphatidylcholine phosphohydrolase (PCase) were used to deplete phospholipid(s) from the particulate fraction and their effects on PDE activity were investigated. 3. Treatment with SMase or PIase did not affect PDE activity whereas treatment with PCase resulted in inhibition. 4. It was also found that the PCase used not only hydrolyzed phosphatidylcholine but also other phospholipids such as phosphatidylethanolamine, phosphatidylserine and sphingomyelin. PMID- 2555230 TI - Evidence for glycerol 3-phosphate:glucose transphosphorylase activity in bloodstream Trypanosoma brucei brucei. AB - 1. Anaerobic glycolysis in intact bloodstream Trypanosoma brucei brucei was studied. 2. Fructose, glucose and mammose were aerobically catabolized at rates of 3.4, 3.0 and 2.5 and anaerobically at rates of 0.38, 2.75 and 2.35 mumol hexose/hr/10(8) trypanosomes respectively. 3. Glycerol 3-phosphate and ADP accumulated approximately to the same level from anaerobic catabolism of the three hexoses. However, fructose catabolism stopped within 15-20 min but addition of glucose to these already immobilized trypanosomes temporarily caused a rapid characteristic drop in glycerol 3-phosphate level at a rate of 40 nmol/min/10(8) trypanosomes and correspondingly glucose 6-phosphate, glycerol and pyruvate levels were raised. 4. These observations are not consistent with the proposed requirements for the reverse glycerol kinase in anaerobic net ATP production. Instead, we propose a glycerol 3-phosphate:glucose transphosphorylase that catalyses the formation of glycerol and glucose 6-phosphate. PMID- 2555232 TI - Endogenous proteolysis of the human erythrocyte membrane as studied by two dimensional gel electrophoresis and electron spin resonance. AB - 1. Endogenous proteolysis in human erythrocyte membranes was studied in human erythrocyte membranes incubated at 37 degrees C by monitoring changes in 2-D electrophoretic pattern of membrane polypeptides and in the spectra of maleimide spin labeled membranes. 2. A strong effect of exogenous proteases derived from contaminating other blood elements was found, resulting in formation of specific spots on 2-D electropherograms, requiring very careful leukocyte removal in investigations of red cell membrane protein composition and proteolysis. 3. Studies of the effects of protease inhibitors and Ca2+ confirmed a complex pattern of endogenous red cell membrane proteolysis ("self-digestion") involving many substrates and enzymes. 4. A promoting effect of high concentrations (150 mM) of Ca2+ on endogenous red cell membrane proteolysis was found. PMID- 2555231 TI - The isolation and partial sequencing of human bone alkaline phosphatase gene. AB - 1. A charon 4A human fetal liver genomic library was screened for human alkaline phosphatase sequences using the cloned human bone cDNA as a hybridization probe. 2. A positive clone was obtained and then characterized by restriction endonuclease cleavage analysis, hybridization experiments and partial DNA sequencing. PMID- 2555233 TI - Insulin-like growth factor-I and insulin increase the stimulatory guanine nucleotide binding protein (Gs) in cultured bovine adrenal cells. AB - The present study shows that pretreatment of BAC cells with insulin or insulin like growth factor-I (IGF-I) enhances the cAMP response to maximal concentrations of ACTH and cholera toxin. However, the effects of IGF-I at a nanomolar concentration (50 ng/ml) were higher than for insulin at the same concentration but similar for insulin at a micromolar concentration (10 micrograms/ml). We have investigated whether the effects of the two peptides can be related to some modifications of the guanine nucleotide regulatory binding protein Gs. Insulin enhanced Gs as observed by ADP ribosylation and immunoblotting but the effects were approximately the same at nanomolar and micromolar concentrations; again, the effects of IGF-I (50 ng/ml) were higher. These results indicate that both IGF I and insulin increase the Gs complex of adenylate cyclase, but IGF-I is more potent than insulin at physiological concentrations. PMID- 2555234 TI - Dictyostelium erasure mutant HI4 abnormally retains development-specific mRNAs during dedifferentiation. AB - The Dictyostelium mutant HI4 progresses through morphogenesis normally, but is defective in the reverse program of dedifferentiation. In contrast to dedifferentiating wild-type cells, HI4 cells retain the capacity to rapidly reaggregate well after the "erasure event" employing a nonchemotactic aggregation mechanism involving random collisions and cohesion. They also do not lose contact sites A (gp80) at the prescribed time in the dedifferentiation program. HI4 cells accumulate transcripts of the cysteine protease gene CP2 (formerly referred to as 16G1) and the cohesion glycoprotein gene gp80 at the correct times in the morphogenetic program, but abnormally retain these transcripts at high levels well after the prescribed times at which they are lost in wild-type cells during the reverse program of dedifferentiation. The retention of these mRNAs in HI4 cells after the erasure event is not due to abnormal maintenance of a high level of intracellular cAMP during dedifferentiation. The rapid reduction in the level of gp80 transcript which can be effected by the addition of cAMP prior to the erasure event in wild-type cells is also retained by HI4 cells well after the erasure event. The results suggest that cells possess at least two mechanisms for the reduction of gp80 transcript. One involves the immediate response to cAMP and may function during the forward program of development. The second functions specifically during the reverse program of dedifferentiation. It is this latter, erasure-specific mechanism which is selectively defective in the HI4 variant. PMID- 2555235 TI - Quantitation of type II topoisomerase in oocytes and eggs of Xenopus laevis. AB - We have generated a monoclonal antibody and a polyclonal antiserum specific for Xenopus laevis topoisomerase II. Using quantitative immunoprecipitation and Western blotting techniques, we have determined the content of topoisomerase II in X. laevis oocytes during oogenesis and in unfertilized eggs. An average stage I oocyte contains 6 pg of topoisomerase II. The content of topoisomerase II per oocyte increases throughout oogenesis to 1.5 ng per stage VI oocyte. The topoisomerase II protein in stage VI oocytes is stored in the germinal vesicles. The cellular content of type II topoisomerase increases significantly when stage VI oocytes are hormonally stimulated to mature into unfertilized eggs. PMID- 2555236 TI - Cytokinin oxidase activity in the cellular slime mold, Dictyostelium discoideum. AB - The cytokinin N6-(delta 2-isopentenyl)adenine (i6Ade) is produced during the development of the cellular slime mold, Dictyostelium discoideum, and functions in this organism as the immediate precursor of the spore germination inhibitor, discadenine. The metabolism of i6Ade in axenic cultures of D. discoideum Ax-3 amoebae has been investigated in the present study. An enzyme activity that specifically catalyzes the degradation of i6Ade has been detected in Ax-3 amoebae. This enzyme is similar to the cytokinin oxidases present in higher plant systems and cleaves the N6-side chain of i6Ade to form adenine. Discadenine synthase activity was also detected in axenically cultured Ax-3 amoebae. The cytokinin oxidase activity detected in Dictyostelium decreased during aggregation and development of Ax-3 amoebae and in starving Ax-3 amoebae maintained under either fast-shake (230 rpm) or slow-shake (70 rpm) conditions. In the latter case, the fall in enzyme activity was accelerated by treatment with cyclic AMP. In contrast to these results, discadenine synthase activity in Ax-3 amoebae rose sharply during the culmination phase of development, exhibited little change in starving Ax-3 amoebae maintained under fast-shake conditions, and fell under slow shake conditions unless the amoebae were treated with cyclic AMP. Possible functions of the Dictyostelium cytokinin oxidase and the significance of the i6Ade metabolism observed in vegetative Dictyostelium amoebae are discussed. PMID- 2555237 TI - Hypertrophy and calcification of rabbit permanent chondrocytes in pelleted cultures: synthesis of alkaline phosphatase and 1,25-dihydroxycholecalciferol receptor. AB - Cartilage calcification at specific sites is a key event that leads to skeletal development and growth. To obtain insights into the control of cartilage calcification, we examined whether cells distributed in permanent cartilage regions might have the ability to express the calcification-related phenotype in a permissive environment. Chondrocytes were isolated from the permanent and growth plate cartilages of 4-week-old rabbit ribs. They were seeded as a pelleted mass in a centrifuge tube and cultured in Eagle's minimum essential medium supplemented with 10% fetal bovine serum. These cells proliferated for several generations, and then synthesized large amounts of proteoglycans, yielding a cartilage-like tissue in 16 days. Cultures from the permanent and growth plate cartilages showed similar time courses for increases in DNA synthesis and proteoglycan production that reached similar maximal levels. Thereafter, they initiated the syntheses of alkaline phosphatase and 1,25-dihydroxycholecalciferol receptor and induced matrix calcification without additional phosphate. The increases in alkaline phosphatase, 1,25-dihydroxycholecalciferol receptor, and calcium contents in cultures from the permanent cartilage were consistently delayed for 4-7 days relative to the growth plate-derived cells, but caught up by Day 28. The maximal levels of alkaline phosphatase and 1,25 dihydroxycholecalciferol receptor in the cultures from the permanent cartilage were 40- to 100-fold higher than that of the in vivo permanent cartilage. These results provide evidence that permanent cartilage cells in postnatal young rabbit ribs have the capacity to express alkaline phosphatase and 1,25 dihydroxycholecalciferol receptor and induce calcification in a permissive environment, although they never express these calcification-related phenotypes in vivo. PMID- 2555238 TI - Blastocoel expansion in the preimplantation mouse embryo: stimulation of sodium uptake by cAMP and possible involvement of cAMP-dependent protein kinase. AB - Elevating cAMP levels in mouse blastocysts increases the rate of blastocoel expansion (F. Manejwala, E. Kaji, and R. M. Schultz, 1986, Cell, 46, 95-103), which requires extracellular sodium (F. Manejwala, E. J. Crago, Jr., and R. M. Schultz, 1989, Dev. Biol. 133, 210-220). We report that cAMP analogs that can activate the cAMP-dependent protein kinase stimulate 22Na+ uptake by cavitating mouse blastocysts and that inhibitors of cAMP-dependent protein kinase activity inhibit the cAMP-stimulated increase in both the rate of blastocoel expansion and 22Na+ uptake. PMID- 2555239 TI - Banting lecture 1989. Structure and function of insulin receptors. AB - The actions of insulin are mediated by an integral plasma membrane protein, the insulin receptor. The processed receptor is a tetramer composed of two alpha subunits that bind insulin and two beta-subunits that traverse the plasma membrane and are, in their cytosolic domains, protein tyrosine kinases. The insulin proreceptor cDNA has been cloned and its complete amino acid sequence deduced. The availability of cDNA permitted an analysis of both the role of protein tyrosine kinase activity in insulin action and the autophosphorylation sites that regulate kinase activity. The human cDNA probe has also been used to identify a putative Drosophila insulin receptor. This work is reviewed, and approaches that may be used to identify physiological substrates for the receptor kinase are suggested. PMID- 2555240 TI - Reduction of glycemic and lipid levels in db/db diabetic mice by psyllium plant fiber. AB - The soluble plant fiber psyllium significantly reduced fasting glucose and total cholesterol levels in the C57BL/KsJ db/db diabetic mouse relative to placebo-fed mice. Insulin levels were significantly higher in psyllium-fed than placebo-fed animals, indicating this fiber may delay the progression of diabetes in the animal model. High-density lipoprotein cholesterol levels rose moderately in both psyllium- and placebo-fed animals during the study, whereas triglyceride levels remained unchanged in both groups. Psyllium's effect on glycemic, lipid, and hormone parameters was not explained by weight loss or reduced food intake; these were similar in psyllium- and placebo-fed animals during the study. Our results show that psyllium fiber can beneficially moderate glycemic and lipid parameters in the db/db diabetes model. PMID- 2555241 TI - Cytoplasmic retinoid-binding proteins and retinoid effects on insulin release in RINm5F beta-cells. AB - Vitamin A (retinol) is required for insulin secretion, and retinoic acid substitutes for retinol in this function. To determine if retinol acts at the beta-cell level, we assayed beta-cells of the rat insulinoma (RINm5F) line for cytosolic retinol- and retinoic acid-binding proteins (CRBP and CRABP) by radioimmunoassay (RIA) and [3H]retinol and [3H]retinoic acid binding to cytosol extracts. Furthermore, we tested whether insulin release from cells was affected by addition of retinol or retinoic acid to culture medium. RINm5F cells were grown to near confluence before assay of CRBP and CRABP. Scatchard analysis showed the Kd for retinol to be approximately 6 nM at a level of 4.5 pmol/mg protein or 300,000 sites/cell. Sucrose density-gradient assay showed single discrete peaks migrating at 2S for both retinol and retinoic acid. RIA of whole cell extracts showed CRBP and CRABP levels of 5.27 +/- 0.41 and 2.95 +/- 0.75 pmol/mg protein, respectively. Retinol (1.75 microM) and retinoic acid (0.175 and 1.75 microM) increased KCl-induced insulin release. Considered together, the presence of CRBP and CRABP in a beta-cell line and the increase in KCl-induced insulin release by retinol and retinoic acid are consistent with the idea that retinol has a functional role in insulin secretion and suggest a potential mechanism of action at the beta-cell level similar to that observed in other retinoid-responsive cells. PMID- 2555242 TI - Flow-cytometric detection of human anti-rat insulinoma antibodies in relation to anti-human islet cell and anti-insulin antibodies. Recognition of distinct antigens by antibodies in early type I diabetes. AB - Flow cytometry was recently introduced for the detection of antibodies in human serum to a cultured insulin-secreting rat insulinoma cell line (RINm5F) to investigate humoral immune reactivity in newly diagnosed type I (insulin dependent) diabetic patients. Fifty-three patients were observed for 6-20 mo after clinical onset of diabetes with a reported duration of symptoms of less than 6 wk. Human anti-RINm5F antibodies were detected in 28%, human anti-islet cell antibodies in 62%, and anti-insulin autoantibodies in 36% of patients before initiation of insulin therapy. Occurrence of human anti-RINm5F antibodies at this stage was correlated with human anti-insulin autoantibodies rather than with the formation of anti-islet cell antibodies. Incidence of anti-RINm5F antibodies in individuals with duration of diabetes greater than 6 wk was 38%, whereas human anti-islet cell antibodies and anti-insulin antibodies became detectable in 72 and 61% of the patients, respectively. These findings are in line with previous reports of immunoprecipitation by human diabetic serums of a 64,000-Mr antigenic structure in freshly prepared rat islet cells. The results suggest a reactivity of distinct classes of antibodies in serums of patients with type I diabetes to disparate antigens on human islet cells and cloned rat insulinoma cells and, moreover, reactivity to insulin as the secreted product. Further characterization of the reacting RINm5F antigens and prospective studies in subjects at risk for diabetes are required to validate the application of RIN cells to the investigation of immune mechanisms involved in the pathogenesis of human type I diabetes. PMID- 2555243 TI - Characterization of endothelin receptors and effects of endothelin on diacylglycerol and protein kinase C in retinal capillary pericytes. AB - Retinal capillary pericyte is a cell type selectively lost in early diabetic retinopathy. The physiological function of pericytes is not yet clearly identified, although it probably has contractile properties. We determined the specific binding of endothelin 1, a 21-amino acid peptide with potent vasoconstrictive action, and the stimulation of diacylglycerol/protein kinase C (DAG/PKC) pathway in cultured retinal capillary pericytes by endothelin. A single specific binding site for 125I-labeled endothelin was identified, with an apparent Kd of 1.3 nM and a maximal binding capacity of approximately 1-2 x 10(5) sites/cell. Endothelin (100 nM) increased total cellular DAG content by 15% at 5 min and 24% at 10 min. When pericytes were labeled isotopically with [3H]glycerol, endothelin stimulated [3H]DAG formation by 100% at 10 min and 88% at 30 min. After 10 min of endothelin treatment, PKC activities were increased by 60 and 100% in the membranous and cytosolic pools, respectively. We conclude that bovine retinal capillary pericytes possess numerous high-affinity specific binding sites for endothelin that mediate the action of endothelin by the stimulation of the DAG/PKC pathway in pericytes. These findings suggest that endothelin is a regulator of the contractile properties of pericytes, which may be adversely affected in diabetic retinopathy. PMID- 2555244 TI - Leukotrienes C4 and D4 potentiate acid production by isolated rat parietal cells. AB - The effects of leukotrienes (LTs) B4, C4, and D4 on acid production by enriched (80%-85%) rat parietal cells were investigated. Acid production was indirectly measured by [14C]aminopyrine uptake into the cells. Leukotriene B4 (10(-10)-10( 6) mol/L) had no effect on basal or prestimulated [14C]aminopyrine uptake. Leukotriene C4 and LTD4 (10(-10)-10(-6) mol/L) also did not change basal acid production but potentiated prestimulated [14C]aminopyrine uptake. Maximal effects were observed with 1 x 10(-7) mol/L LTC4 or with 3 x 10(-7) mol/L LTD4. At these concentrations LTC4 and LTD4 induced the indicated increases above the responses to the following prestimulants (= 100%): 10(-4) mol/L histamine (71% and 74%, respectively), 10(-5) mol/L forskolin (54% and 106%), 10(-4) mol/L dibutyryl cyclic adenosine monophosphate (34% and 81%), and 10(-4) mol/L carbamylcholine (160% and 116%). Yet, adenosine triphosphate (2.5-5 x 10(-3) mol/L)-induced [14C]aminopyrine uptake in digitonin-permeabilized parietal cells was not further increased by LTC4 or LTD4. At 10(-5) mol/L the selective LTD4 antagonist L 660,711 (MK-571) reduced the effect of 3 x 10(-7) mol/L LTD4 by 74% but had no effect on the potentiation by LTC4. We conclude that the sulfidopeptide LTs C4 and D4, but not LTB4, exert a direct effect on rat parietal cells, and that this effect seems to be mediated by separate specific receptors. Leukotriene C4 and LTD4 potentiate prestimulated H+ formation by interacting with an intracellular mechanism that is commonly activated upon occupation of histamine H2- as well as muscarinic receptors, and that is also activated by the postreceptor stimuli forskolin and dibutyryl cyclic adenosine monophosphate; yet, this mechanism seems to be localized proximal to the H+,K+-adenosine triphosphatase. PMID- 2555245 TI - Ultrasonic Doppler studies of hepatocellular carcinoma and comparison with other hepatic focal lesions. AB - One hundred fifty-four liver lesions, including 63 hepatocellular carcinomas, were studied to determine the value of duplex ultrasound in the diagnosis of small hepatocellular carcinomas. Arterial Doppler signals were obtained either within the body of the tumor, at its periphery, or in both locations, from 28 to 37 hepatocellular carcinomas less than or equal to 3 cm in diameter and from all 26 hepatocellular carcinomas with a diameter greater than 3 cm. Arterial Doppler signals were obtained at the periphery of 5 of 7 cholangiocarcinomas, 4 of 11 liver metastatic tumors, and 5 of 23 hemangiomas. No such signals were obtained from 29 regenerative nodules, 10 hepatic pseudotumors, and 11 liver cysts. The mean peak systolic frequency seen in hepatocellular carcinoma (1.2 kHz) was significantly greater than in cholangiocarcinoma (0.6 kHz), metastatic tumors (0.5 kHz), or hemangiomas (0.3 kHz). A peak systolic frequency of greater than 3 kHz was found in 6 of 8 hepatocellular carcinomas greater than or equal to 4 cm in diameter with angiographically proven arterioportal shunting, whereas the value in other hepatocellular carcinomas or other hepatic focal lesions was less than 2.6 kHz. This study showed that the peak systolic shift was related to the degree of arterioportal shunting. Because shunting is either minor or nonexistent in small hepatocellular carcinomas, the value of duplex Doppler ultrasound in the diagnosis of these lesions appears to be limited. PMID- 2555246 TI - [HIV infections and cervical neoplasms]. AB - Between 1985 and 1989 108 HIV-positive patients underwent cytologic examinations at the Dept. of Obstetrics and Gynecology, Univ. of Berlin. These patients showed 15 times more pathologic smears (group IIId to V) than all other patients of the hospital investigated. By histological controls 10 cervical intraepithelial neoplasias (CIN I to III) and 5 invasive carcinomas were found. A significantly high percentage (40.3%) of the smears showed signs of a papilloma-virus infection, even the pathologic smears in a percentage of 80.7%. PMID- 2555247 TI - Thyrotropic activity of the ovine corticotropin-releasing factor in the chick embryo. AB - An injection of several doses of ovine corticotropin-releasing factor (oCRF) was made into an allantoic blood vessel of 18-day-old chick embryos. All doses used (0.5, 1, 2, and 5 micrograms) induce a quick increase of plasma corticosterone concentrations after 15 and 30 min and 1 and 2 hr and this in a dose-dependent manner with the high doses having a prolonged effect lasting up to 2 hr. An increase in plasma levels of thyroxine (T4) and triiodothyronine (T3) was observed after 1 and 2 hr. After the oCRF injection, no stimulation of the hepatic 5'-monodeiodination activity was observed and there was no increase, but even a small decrease in the T3 to T4 ratio could be calculated. It was therefore concluded that oCRF in the chick embryo has a thyrotropic effect presumably by stimulating the release of thyrotropin from the pituitary. PMID- 2555248 TI - Distribution of renal calcitonin binding sites in mammalian and nonmammalian vertebrates. AB - Calcitonin (CT), the hypocalcemic hypophosphatemic hormone, is present in many vertebrate species. The principal target organs for CT are, in mammals, kidney and bone, and in fish, bone and gill. We have investigated the presence of renal calcitonin binding sites in a fish (Salmo gairdneri), two amphibians (Bufo bufo and Rana esculenta), two reptiles (Pseudemys scripta and Gekko gecko), and two birds (Gallus domesticus and Cothurnix japonica). We compared their distribution to a mammal, the rat (Wistar strain), using quantitative autoradiography methods. Moreover, CT binding sites were also characterized in the chicken kidney by a membrane assay. No renal 125I-sCT binding sites were detected in the fish, the amphibians, and the reptiles studied. In the rat, binding sites were present in the outer medulla and in the cortex. In the chicken and in the quail, scattered binding sites were also observed in the medulla and in the cortex, whose pattern seemed to follow the distribution of the glomeruli and/or the collecting tubules. Chicken kidney membranes were also purified by sucrose gradient centrifugation. Scatchard analysis of the binding data revealed the presence of one type of 125I sCT binding site with an apparent dissociation constant of 4.3 nM and a number of sites of 73 fmol/mg/protein. The present results suggest that calcitonin renal receptors appeared late in evolution and thus that a regulation of renal function by calcitonin was only effective in birds. PMID- 2555250 TI - Autotransplantation of avian parathyroid glands: an animal model for studying parathyroid function. AB - The parathyroid glands of chickens were autotransplanted and the return of parathyroid function following transplantation was determined. Parathyroidectomy (PTX) resulted in a marked hypocalcemia (5.2 +/- 0.2 mg/dl) 4 hr following PTX. Plasma calcium (PCa) had declined to 4.3 +/- 0.2 mg/dl 24 hr after PTX. Parathyroid glands were transplanted subcutaneously 24 hr after removal and 24 hr later, PCa had risen to 8.6 +/- 0.5 mg/dl. Seven days after PTX. PCa increased to 10.3 +/- 0.2 mg/dl and by 14 days was indistinguishable from control levels (10.8 +/- 0.2 mg/dl vs. 11.0 +/- 0.2 mg/dl, respectively). When chicks with transplanted glands were fed a low Ca (0.08%) diet for 2 weeks they were able to maintain plasma PCa at levels comparable to control birds. Removal of the transplanted glands resulted in marked decreases in PCa (from 9.7 +/- 0.3 to 5.6 +/- 0.8 mg/dl), in the fractional excretion of phosphate, in urine cAMP, and in renal 25OH-vitamin D3-1 alpha-hydroxylase activity. Stepwise reductions in PCa and 1 alpha-hydroxylase activity were produced in partially PTX and fully PTX chicks by removing part or all of the parathyroid tissue. These data suggest that the transplanted parathyroid tissue was the major source of circulating PTH and that it may be possible to produce different degrees of acute hypoparathyroidism by varying the amount of transplanted parathyroid tissue removed surgically. Chickens with transplanted parathyroid glands thus provide a convenient animal model in which to study parathyroid function in an avian species. PMID- 2555249 TI - Melatonin reduction by lithium and albinism in quail and hamsters. AB - Melatonin was measured by radioimmunoassay in several genetic strains of Japanese quail. Plasma melatonin (PM), measured at the nighttime peak, was highest in wild type quail reared on a diurnal lighting schedule; this PM peak was suppressed in continuous light. Albino quail had low melatonin levels, whether reared under diurnal conditions or in continuous light. Ocular melatonin was also suppressed in albinos and in dilute mutants. At midday sampling, melatonin was only half as high in albinos as in wild types. Intraocular pressure (IOP, daytime readings) was uniformly low in diurnal birds and was elevated in all quail reared under continuous light. Thus in pigmented birds, a high melatonin level was associated with high IOP, but in albinos displaying high IOP, ocular melatonin was not elevated. Lithium chloride, mixed in the feed, brought about a pronounced reduction in plasma, pineal, and ocular melatonin, in wild-type quail reared on a diurnal schedule. This confirms earlier findings in lithium-fed rats. Golden hamsters displayed a characteristic diurnal cycle of pineal melatonin, with a sharp middark peak; in albino hamsters, also kept on a diurnal schedule, this peak occurred at the same time, but albinos had melatonin levels only about one third as high as those of pigmented animals. PMID- 2555251 TI - A simple procedure for detecting proteins that bind preferentially to kDNA networks. AB - A procedure is described in which a fractionated protein extract from the trypanosome Crithidia fasciculata was incubated either with deproteinized C. fasciculata kDNA networks or nuclear DNA. At least three proteins in the range of 45-65 kDa had a specific affinity for the networks. We suggest that these proteins may play an important role in the biology of the kinetoplast. PMID- 2555252 TI - Ty1 transposition in Saccharomyces cerevisiae is nonrandom. AB - A large collection of Ty1 insertions in the URA3 and LYS2 loci was generated using a GAL1-Ty1 fusion to augment the transposition frequency. The sites of insertion of most of these Ty elements were sequenced. There appears to be a gradient of frequency of insertion from the 5' end (highest frequency) to the 3' end (lowest frequency) of both loci. In addition we observed hotspots for transposition. Twelve of the 82 Ty1 insertions in the URA3 locus were inserted in exactly the same site. Hotspots were also observed in the LYS2 locus. All hotspots were in the transcribed part of the genes. Alignment of the sites of insertion and of the neighboring sequences only reveals very weak sequence similarities. PMID- 2555253 TI - Physical and functional definition of the Drosophila Notch locus by P element transformation. AB - Notch is a developmentally regulated locus which controls the differentiation of various Drosophila tissues, among them the embryonic nervous system. Molecular analysis has suggested that Notch is defined by an approximately 40-kb transcription unit which is spliced into a 10.2-kb mRNA composed of nine exonic regions and coding for a 2703-amino acid long transmembrane protein that shows homology to the mammalian epidermal growth factor. Here, we define the 5' end of the transcription unit and determine the sequences deleted in a Notch mutation involving the 5' nontranscribed region. Using a Notch cosmid vector we demonstrate by P element-mediated transformation that all sequences necessary for Notch function are confined in an approximately 40-kb long genomic region. cDNA sequences are used to construct a 15-kb "minigene" which lacks most, but not all, introns and its functionality is also tested by P element transformation. We show that, unlike the cosmid vector which is capable of rescuing completely all Notch mutations, only certain phenotypes can be rescued by the "minigene." The functional implications of our findings are discussed. PMID- 2555254 TI - The P-M and the 23.5 MRF (hobo) systems of hybrid dysgenesis in Drosophila melanogaster are independent of each other. AB - Strains of Drosophila melanogaster bearing the male recombination factor 23.5 MRF induce hybrid dysgenesis in a way which is highly reminiscent of the P-M system, and, most probably, causally related to the activity of the transposable element hobo. We have investigated potential interactions between the two systems of hybrid dysgenesis by studying mixed lines derived from bidirectional crosses between 23.5 MRF and P strains, and analyzed their potentials to induce or suppress the occurrence of dysgenesis. All new lines possess the P induction abilities, as determined by two different procedures, and have also acquired a P cytotype. In contrast, some of them lost their ability to induce the non-P-M dysgenesis, as well as to suppress the action of 23.5 MRF. This loss of the 23.5 MRF induction abilities parallels the selective loss of full-length hobo elements from the genome of these lines, providing further substantiation to the notion that the 23.5 MRF activity is directly linked to this transposable element. PMID- 2555256 TI - [Detection of new genes participating in the trans-regulation of locus yellow of MDG-4 in Drosophila melanogaster]. AB - The aim of the present work was to obtain mutations in the genes involved in regulation of the yellow locus and mdg4. For this purpose, we searched for mutations changing phenotypic expression of the y(2) mutation induced by mdg4 insertion into the regulatory region of the yellow locus. Mutations have been obtained in the earlier described system of prolonged genome instability, sometimes combined with P-M hybrid dysgenesis. The mutation mod(mdg4) in a novel gene, modifier of mdg4, was detected which either enhanced or suppressed a phenotypic expression of several mutations induced by mdg4 insertion. We suggest that mod(mdg4) controls the expression of mdg4. In addition, the mutations in five loci located on the X chromosome have been found which enhanced the mutation phenotype of several y alleles induced by insertions of different mobile elements in the regulatory region of the latter. Possibly, the protein products of these genes designated as enhancers of yellow-1, 2, 3, 4 and 5 are directly or indirectly involved in control of the yellow locus expression. PMID- 2555255 TI - Molecular analysis of a transposon-induced deletion of the nivea locus in Antirrhinum majus. AB - The transposable element Tam3 of Antirrhinum majus is capable of causing large scale chromosomal restructuring. It induced a large deletion at the nivea locus, to produce the allele niv-:529. The deletion removed the entire nivea coding region while the element remains intact with the potential to induce further rearrangements. Genetic experiments showed that the endpoint of the deletion (called x) is closely linked to nivea. The DNA sequences of niv-:529, a genomic excision of Tam3 from niv-:529, and the original genomic position of x have been determined. These data suggest that the deletion could have resulted from an abortive transposition or through breakage and religation. PMID- 2555257 TI - [Antibodies in dogs to the virus of tick-borne encephalitis (early summer encephalomyelitis/tick-borne encephalitis) in Greece]. AB - A FSME (tick-borne-encephalitis) epidemiological study among dogs undertaken in Greece, revealed in a series of examinations 0.97% of the 206 dogs with antibodies against FSME (tick-borne-encephalitis). In an additional study of 429 dogs 8.6% reacted positive. PMID- 2555258 TI - Vectors permitting visual monitoring of simple transposition events. AB - The construction and use of two novel transposon(Tn)-delivery vectors is described. These vectors carry Inc.W or Inc.N broad-host-range transfer functions cloned next to the narrow-host-range replicon of pBR329. The host specificities of pSLX10 and pSLX23 both complement and extend the host specificities of existing Tn delivery vectors. Plasmids pSLX10 and pSLX23 were shown to transfer at high frequency in intergeneric matings. The lux genes which are present on each vector permit the visual monitoring of transconjugants which have retained a Tn element, but are devoid of plasmid molecules. pSLX10 and pLSX23 were efficiently used to generate a range of auxotrophic mutants in various strains of Pseudomonas as well as to clone genes from Serratia liquefaciens. These vectors may have general applicability to identify and clone genes in a wide range of Gram-negative bacteria. PMID- 2555259 TI - Nucleotide sequence of the Rickettsia prowazekii ATP/ADP translocase-encoding gene. AB - The Rickettsia prowazekii ATP/ADP translocase (Tlc) gene (tlc), previously cloned in Escherichia coli was localized to a 1.6-kb chromosomal fragment. Nucleotide sequence analysis of this fragment revealed an open reading frame of 1494 bp that could encode a hydrophobic protein of 497 amino acids (aa) with an Mr of 56,668. Analysis of the deduced aa sequence revealed that it contained twelve potential membrane-spanning regions. Comparisons between the deduced aa sequence of the R. prowazekii ATP/ADP Tlc and the sequences of mitochondrial (mt) Tlc revealed no detectable homologies between the rickettsial and mt sequences. The major protein synthesized in E. coli minicells containing the rickettsial gene exhibited and Mr of approx. 34,000. PMID- 2555260 TI - Simultaneous synthesis and assembly of various hepatitis B surface proteins in Saccharomyces cerevisiae. AB - Yeast transposon of class-1-based vectors, allowing integration at a series of chromosomal loci by homologous recombination with resident transposons, were constructed. Using such vectors, we have introduced several copies of an expression cassette encoding the major hepatitis B surface protein as well as expression cassettes encoding the middle (M) or/and the large (L) surface protein into Saccharomyces cerevisiae. In extracts of such strains, coassembly of the different proteins into a single lipoprotein structure is observed. This was demonstrated by immunoprecipitation of the major protein using monoclonal antibodies directed specifically against epitopes that are present only on the M or the L protein. These results show that hepatitis B surface antigen envelope proteins synthesized in yeast are able to assemble into structures composed of different polypeptides. This opens the possibility of producing in yeast a variety of particles carrying well-defined amounts of preS epitopes on their surface. Also, one can envisage the production of mixed particles containing different foreign epitopes on their surface, in defined relative abundance, which could be useful for vaccine applications. PMID- 2555261 TI - Mobile introns: definition of terms and recommended nomenclature. AB - A number of introns in mitochondrial, chloroplast, nuclear or prokaryotic genes have recently been shown to encode double-strand sequence-specific endonucleases. Such introns are mobile genetic elements that insert themselves at or near the cleaved sites. A uniform nomenclature to designate the molecular elements involved in the phenomenon of intron mobility is proposed. PMID- 2555262 TI - A site-specific endonuclease and co-conversion of flanking exons associated with the mobile td intron of phage T4. AB - The product of the td intron open reading frame (ORF) of phage T4 is required for high-frequency transfer of the intervening sequence from intron-plus (In+) to intron-minus (In-) alleles. In vivo studies have demonstrated that the td ORF product targets cleavage of td In- DNA, and that cleavage is correlated with intron inheritance [Quirk et al., Cell 56 (1989) 455-465]. In the present study we show by in vitro synthesis of the td intron ORF product, that the protein possesses endonuclease activity and efficiently cleaves double-stranded DNA at or near the site of intron integration. In addition, we demonstrate that intron insertion is accompanied by co-conversion of the flanking exon sequences. Co conversion of markers within 50 nt surrounding the site of intron insertion occurred at a high frequency (80-100%), and decreased at greater distance from the intervening sequence. Co-conversion may provide a mechanism for maintaining exon-intron RNA contacts required for accurate splicing of the relocated intron. Cleavage of target DNA by an intron endonuclease and co-conversion of flanking exon sequences are both features associated with mobile introns of eukaryotes, indicating a common mechanism for intron transfer in the eukaryotic and prokaryotic kingdoms. PMID- 2555263 TI - The splicing of maize transposable elements from pre-mRNA--a minireview. AB - There are six examples of maize transposable elements that are spliced from pre mRNA. These represent the first introns that have been added to nuclear genes in recent years. All six are members of the Ac/Ds or Spm/dSpm family of elements and all have been inserted into exons of active genes within the last twenty-five years. The structure of these element-introns and the sequences involved in their splicing are presented. These examples illustrate how active transposable elements can also function as introns and how they may evolve into stable introns. PMID- 2555264 TI - Group I introns as mobile genetic elements: facts and mechanistic speculations--a review. AB - Group I introns form a structural and functional group of introns with widespread but irregular distribution among very diverse organisms and genetic systems. Evidence is now accumulating that several group I introns are mobile genetic elements with properties similar to those originally described for the omega system of Saccharomyces cerevisiae: mobile group I introns encode sequence specific double-strand (ds) endoDNases, which recognize and cleave intronless genes to insert a copy of the intron by a ds-break repair mechanism. This mechanism results in: the efficient propagation of group I introns into their cognate sites; their maintenance at the site against spontaneous loss; and, perhaps, their transposition to different sites. The spontaneous loss of group I introns occurs with low frequency by an RNA-mediated mechanism. This mechanism eliminates introns defective for mobility and/or for RNA splicing. Mechanisms of intron acquisition and intron loss must create an equilibrium, which explains the irregular distribution of group I introns in various genetic systems. Furthermore, the observed distribution also predicts that horizontal transfer of intron sequences must occur between unrelated species, using vectors yet to be discovered. PMID- 2555265 TI - Electroporation-mediated transformation of lysostaphin-treated Clostridium perfringens. AB - A reliable and efficient method has been developed for the electroporation mediated transformation of Clostridium perfringens with plasmid DNA. Transformation of vegetative cells of C. perfringens strain 13 with the 7.9-kb Escherichia coli-C. perfringens shuttle plasmid pHR 106 required pretreatment with lysostaphin (2 to 20 micrograms/ml) for 1 h at 37 degrees C. Cells harvested early in the logarithmic stage of growth were transformed more efficiently than cells at other growth phases. The transformation frequency increased with the DNA concentration, to a saturating level at 5 to 10 micrograms DNA/ml. The transformation frequency was proportional to the field strength and time constant of the electroporation pulse; however, the field strength was a far more important parameter. A cell density between 1 x 10(8) and 5 x 10(8) cells/ml proved to be optimal for transformation. The procedure was capable of generating up to 3.0 x 10(5) transformants per micrograms DNA. The potential value of the method for the cloning of C. perfringens genes was demonstrated by the cloning of the clostridial tetracycline-resistance determinant, tetP, from the E. coli recombinant plasmid pJIR71, into C. perfringens strain 13. PMID- 2555267 TI - Expression of the adenyl cyclase-encoding gene (cya) of Rhizobium meliloti F34: existence of two cya genes? AB - To gain insight into the role of cyclic AMP (cAMP) in Gram-negative soil bacteria, we have studied the expression of an adenyl cyclase-encoding gene 'cya' of Rhizobium meliloti F34. In both Escherichia coli and Bradyrhizobium japonicum, the gene is expressed from a promoter(s) contained on a 2.6-kb fragment of the cloned insert, which may indicate the presence of a functional 'cya' promoter or the coincidental presence of sequences which can function as promoters in these two species. The study of 'cya'-lac fusion activity in R. meliloti indicated that the 'cya' gene is not expressed at detectable levels and, thus, may not contribute to the modulation of cAMP levels under the growth conditions employed. R. meliloti strains carrying defined genomic mutations at the 'cya' locus were still capable of the synthesis of near wild-type levels of cAMP. These results suggest that the cloned 'cya' gene is not the key determinant responsible for cAMP synthesis under the culture conditions employed. PMID- 2555266 TI - Homologous potyvirus and flavivirus proteins belonging to a superfamily of helicase-like proteins. AB - Comparison of the nucleoside triphosphate-binding motif(NTBM)-containing proteins of two groups of apparently distantly related positive-strand RNA viruses (potyvirus and flavivirus), revealed significant sequence similarity. In addition, these two groups of viral proteins show amino acid motifs in common with those conserved in a group of five NTBM-containing proteins from prokaryotic and eukaryotic cells, some of which have been experimentally related to helicase activity. Here we propose that the proteins mentioned above constitute a superfamily of helicase-like proteins, distinct from the one previously described [Gorbalenya et al., FEBS Lett. 235 (1988) 16-24; Hodgman, Nature 333 (1988) 22 23; 578], which includes the NTBM-containing proteins from another group of positive-strand RNA viruses, the 'Sindbis-like' viruses. PMID- 2555268 TI - Promoter mapping and nucleotide sequence of the bchC bacteriochlorophyll biosynthesis gene from Rhodobacter capsulatus. AB - Because there are not yet direct assays for most of the proteins required for differentiation of Rhodobacter capsulatus cytoplasmic membrane into photosynthetically competent intracytoplasmic membrane, a molecular inquiry into the mechanism and regulation of this process is difficult. We have, therefore, chosen to isolate R. capsulatus photosynthesis genes by creating in-frame fusions to lac'Z vectors, and selecting for those that direct appropriately regulated levels of beta-galactosidase in R. capsulatus. One lacZ fusion isolate was used to identify an open reading frame (ORF) of unknown function and flanking sequences that promoted initiation of transcription. The chromosomal copy of this ORF was mutated by insertion of a kanamycin-resistant cartridge into the cloned fragment and substitution for the chromosomal copy by homologous recombination. The phenotype of the resultant mutant cells showed that the ORF encodes 2 desacetyl-2-hydroxyethyl bacteriochlorophyllide a dehydrogenase, an enzyme that catalyzes the penultimate step in bacteriochlorophyll a biosynthesis. The nucleotide sequence of this bchC gene and its 5' regulatory region were determined. The deduced amino acid sequence shows that the bchC gene encodes a 33 kDa protein that is less hydrophobic than integral membrane proteins of R. capsulatus, although there are hydrophobic segments that could in principle interact with a lipid membrane. Results of S1 nuclease protection and primer extension experiments show that a 5' mRNA end is positioned within the cloned segment, and that this 5' end maps to sequences with significant sequence similarity to the previously characterized puf operon promoter region. PMID- 2555269 TI - Cloning and characterization of the acyl-coenzyme A: 6-aminopenicillanic-acid acyltransferase gene of Penicillium chrysogenum. AB - A gene, aat, encoding acyl-CoA: 6-aminopenicillanic acid acyltransferase (AAT), the last enzyme of the penicillin (Pn) biosynthetic pathway, has been cloned from the genome of Penicillium chrysogenum AS-P-78. The gene contains three introns in the 5'-region and encodes a protein of 357 amino acids with an Mr of 39,943. It complements mutants of P. chrysogenum deficient in AAT activity. The aat gene is expressed as a 1.15-kb transcript and the encoded protein appears to be processed post-translationally into two nonidentical polypeptides of 102 and 255 aa, with Mrs of 11,498 and 28,461, respectively. Three proteins of 40, 11, and 29 kDa (the last one corresponding to the previously purified AAT), were identified in extracts of P. chrysogenum. The aa sequence of the N-terminal end of the 11-kDa polypeptide matched the nucleotide (nt) sequence of the 5'-region of aat. The N terminal end of the 29-kDa polypeptide corresponded to the sequence beginning at nt position 916 of the sequenced DNA fragment (nt 441 of aat gene). The aat gene of P. chrysogenum resembles the genes encoding Pn acylases of Escherichia coli, Proteus rettgeri and Pseudomonas sp., all of which encode two nonidentical subunits derived from a common precursor, encoded by a single open reading frame. PMID- 2555270 TI - Site-directed transcription initiation with a mobile promoter. AB - A method is described for the high-level transcription of any DNA segment using bacteriophage RNA polymerases (RNAPs). A synthetic mobile promoter with a template-complementary 3' extension is ligated to the target sequence of interest. Transcription with an appropriate RNAP results in an amplification of approx. 70-fold. In the presence of heterologous DNA, bacteriophage RNAPs are shown to be specific for their cognate mobile promoters. PMID- 2555271 TI - The size and a physical map of the chromosome of Haemophilus parainfluenzae. AB - The physical map of the Haemophilus parainfluenzae chromosome is circular and approx. 2340 kb in circumference. The size of the map was determined by digesting agarose-immobilized chromosomes with the restriction enzymes, NotI (GCGGCCGC), RsrII (CGGATCCG) and ApaI (GGGCCC), and using field-inversion gel electrophoresis to resolve the resulting fragments. The enzymes digest the H. parainfluenzae genome into 7, 10, and 18 fragments, respectively. The map order of the fragments was obtained by using Southern-blot hybridization to establish overlapping regions. PMID- 2555272 TI - Diminished diurnal secretion of corticosterone in aging female but not male rats. AB - Since very little is known about the impact of aging on adrenocortical function in female rats, it was of interest to compare the age changes in the circulating levels of adrenocorticotropin (ACTH) and corticosterone in male and female Sprague-Dawley rats, and correlate these changes with those in plasma prolactin (Prl), which is known to stimulate adrenal steroid secretion. Hormones were measured in young (3-4 months) and old (24-26 months) male rats as well as in young (3-4 months), old (25 months) and senescent (33-35 months) female rats. Sequential plasma samples were obtained from chronically cannulated animals every 30 min from 11:30 a.m. to 4:30 p.m. ACTH was measured in trunk serum--obtained between 11:30 a.m. and 1:30 p.m.--whereas corticosterone and prolactin were measured in plasma using specific RIAs for the corresponding hormones. No age changes were detected in serum ACTH in either sex. The integrated values of plasma corticosterone did not change with age in males but decreased significantly (p less than 0.05) in old and senescent as compared to young females. Plasma corticosterone was higher in young females than in young males but this sex-related difference disappeared with age. Plasma Prl increased significantly with age in both male (p less than 0.05) and female (p less than 0.001) rats but showed no significant correlation with corticosterone levels. The present results suggest that ACTH secretion does not show major age- or sex related changes. Our corticosterone data are compatible with the idea that gonadal aging has a significant impact on plasma glucocorticoid regulation in female rats but not in males. PMID- 2555273 TI - Reactions of adriamycin with microsomal iron and lipids. AB - Iron plays a central role in oxidative injury, reportedly because it catalyzes superoxide- and hydrogen peroxide-dependent reactions yielding a powerful oxidant such as the hydroxyl radical. Iron is also thought to mediate the cardiotoxic and antitumour effects of adriamycin and related compounds. NADPH-supplemented microsomes reduce adriamycin to a semiquinone radical, which in turn re-oxidizes in the presence of oxygen to form superoxide and hence hydrogen peroxide. During this redox cycling membrane-bound nonheme iron undergoes superoxide dismutase- and catalase-insensitive reductive release. Membrane iron mobilization triggers lipid peroxidation, which is markedly enhanced by simultaneous addition of superoxide dismutase and catalase. The results indicate that: i) lipid peroxidation is mediated by the release of iron, yet the two reactions are governed by different mechanisms; and ii) oxygen radicals are not involved in or may actually inhibit adriamycin-induced lipid peroxidation. Microsomal iron delocalization and lipid peroxidation might represent oxyradical-independent mechanisms of adriamycin toxicity. PMID- 2555274 TI - The action of 4-hydroxynonenal on heat shock gene expression in cultured hepatoma cells. AB - Hep G2 cells do not synthesize heat-shock proteins when incubated with ADP-iron, under conditions that can trigger lipoperoxidation. However, exposure of these cells to added 4-hydroxynonenal (HNE), one of the main products of lipoperoxidation, induces the synthesis of hsp70, the most conserved among heat shock proteins. HNE acts mainly on transcription: in Hep G2 cells the increase in the steady-state level of hsp70 mRNA is detectable by two different hybridization techniques. PMID- 2555275 TI - Oxygen radicals acting as chemical messengers: a hypothesis. AB - Based on a critical reappraisal of the reactions of radicals in a biological milieu, a hypothesis is proposed according to which superoxide anion radicals act as biological messengers rather than as mediators or precursors of cellular damage under oxidative stress conditions. PMID- 2555276 TI - Redox cycling in MCF-7 human breast cancer cells by antitumor agents based on mitozantrone. AB - In a series of hydroxyethylaminoalkylaminoanthraquinones (AQ's) based on mitozantrone, 1-AQ (340%) and 1,8-AQ (137%) stimulated basal rate NADPH oxidation (72 + 18 pmol min-1 mg S9 protein-1) whilst 1,4-AQ, 1,5-AQ and mitozantrone had no effect. A similar trend was observed for O2.- generation (measured as nmol acet. cyt c reduction min-1 mg protein-1) by these compounds in MCF-7 S9 fraction: 1-AQ (9.5) and 1,8-AQ (7.9), whilst 1,5-AQ, 1,4-AQ and mitozantrone showed no significant effect. All the AQ's including mitozantrone were cytotoxic to MCF-7 cells in a dose dependent manner with EC50 values as follows: 1-AQ (0.01 micron) greater than doxorubicin (0.4 microM) greater than mitozantrone (0.6 microM) greater than 1,8-AQ (2.0 microM) greater than 1,5-AQ (4.0 microM) greater than 1,4-AQ (8.0 microM). Thus the redox active AQ's were also the most cytotoxic. Mitozantrone however was not redox active but was more cytotoxic than all but 1-AQ hence it would appear that factors other than free radical generation contribute to the antitumor activity of this group of compounds. PMID- 2555277 TI - Free radicals and lipid peroxidation in liver of rats kept on a diet devoid of choline. AB - Rodents kept on a choline devoid (CD) diet up to 14 months develop hepatic lesions progressing through two broad stages. The first is characterized by severe steatosis and increase in cell turnover, the second by a gradual clearance of the deposited fat and fibrosis. Hepatocellular carcinomas eventually arise in rats fed for over 12 months, even though the animals aer not exposed to chemical carcinogens. It has been suggested that the diet may trigger generated thereby may be responsible for initiation of liver cancer and promotion. The radicals would lead to DNA damage, and the altered DNA in a proliferating liver would result in initiation of the carcinogenic process. In this communication we present evidence that the diet used in the above studies contained stable fatty acid isomers with conjugated dienes, which are absorbed and deposited in rat liver. This finding cast doubts on whether a CD diet does indeed cause a peroxidation of cellular membrane lipids. Electron spin resonance (ESR) spectroscopy was also used to investigate whether any abnormal pattern of free radicals exists in the liver of rats fed a CD diet. No significant differences were noted in ESR spectra of either transition metal-centered signals, or organic free radicals. PMID- 2555278 TI - Direct detection of radical production in the ischaemic and reperfused myocardium: current status. AB - Electron spin resonance (e.s.r.) spectroscopy, which is a specific method for directly detecting free radicals in biological systems, has now been used in a number of studies to examine free radical generation in both ischaemic and reperfused myocardial tissue. This review critically assesses the information which has been obtained to date with particular reference to the elucidation of the nature and source of the radicals observed in these studies. PMID- 2555279 TI - Spin trapping of free radicals produced in vivo in heart and liver during endotoxemia. AB - Studies using free radical scavengers and measurements of lipid peroxidation have suggested that free radicals are generated during endotoxemia. Conclusions from these studies have implied that free radicals may participate in the sequence of pathologic events following endotoxin challenge in the experimental animal. Current inferences of free radical generation and involvement have been derived from indirect evidence and are therefore inconclusive. To quantitate the generation of free radicals in vivo during endotoxemia this study employed the use of electron paramagnetic resonance spectroscopy (EPR) combined with spin trapping techniques. Five minutes before intraperitoneal endotoxin administration, trimethoxy-a-phenyl-t-butyl-nitrone [(MeO)3 PBN] was administered intraperitoneally. Experimental animals were always matched with control animals receiving no endotoxin. At either five minutes or twenty-five minutes following endotoxin administration animals were decapitated and hearts and livers were rapidly taken for lipid extraction and EPR evaluation. Analysis of the EPR spectra revealed hyperfine splitting constants that indicated the presence of carbon-centered radical spin adducts in both organ tissues from animals exposed to endotoxin for twenty-five minutes. No signals were present in hearts and livers taken five minutes after endotoxin administration. EPR evaluation did not indicate spin adduct formation in control tissue. These data directly demonstrate that activation of processes in vivo involving free radical generation occur early during endotoxemia, but are not detectable immediately after the endotoxin challenge. PMID- 2555280 TI - Human red cells enhance the formation of 5-lipoxygenase-derived products by neutrophils. AB - Upon activation, human neutrophils generate 5-lipoxygenase products which are involved in inflammation as well as other physiological and pathophysiological processes. We have examined the influence of red cells on the generation of lipoxygenase-derived products by neutrophils utilizing high pressure liquid chromatography system which permitted quantitation of 5-HETE, leukotriene B4 (and its isomers) and the omega oxidation products of leukotriene B4 (20 hydroxyleukotriene B4, 20-carboxyleukotriene B4) within the same sample. Co incubation of red cells with neutrophils (50:1, red cells:neutrophils) resulted in a 722 percent increase in 5-hydroxyeicosatetraenoic acid production and a slight increase in leukotriene B4 and its omega oxidation products which were not accompanied by increases in 15-hydroxyeicosatetraenoic acid production. The role of the sulfhydryl status of the red cell and its ability to scavenge hydrogen peroxide were assessed in relationship to the interaction of red cells on the neutrophil-derived lipoxygenase products. Together, these findings indicate that red cells can regulate the levels of lipid-derived mediators produced by neutrophils. Moreover, they suggest that red cell-neutrophil interactions may be of importance in inflammatory reactions. PMID- 2555281 TI - 15-Hydroxyeicosatetraenoic acid inhibits superoxide anion generation by human neutrophils: relationship to lipoxin production. AB - Human neutrophils can aggregate, degranulate, and release mediators of inflammation including oxygen radicals and lipoxygenase (LO)-derived products of arachidonic acid. The regulation of 5- and 15-lipoxygenases appears to be important since their products (e.g. leukotrienes and lipoxins) display unique spectra of bioactions. Addition of 15-HETE, a product of the 15-LO, to neutrophils in suspension dramatically shifted the LO products generated and led to a dose-dependent increase in lipoxins, while the production of leukotriene B4 and its omega-oxidation products (i.e. 20-COOH-LTB4 and 20-OH-LTB4) was inhibited. Exogenous 15-HETE also dose-dependently inhibited the generation of superoxide anions induced by either the chemotactic peptide f-met-leu-phe or the divalent cation ionophore A23187. Neither lipoxin A4 nor lipoxin B4 (10(-8)-10( 6) M) inhibited O2-. generation induced by either f-met-leu-phe or A23187. These results indicate that in addition to serving as a substrate for lipoxin generation, 15-HETE also inhibits superoxide anion generation by human neutrophils. Together they provide further evidence to suggest that products of the 15-lipoxygenase may serve a regulatory role at inflammatory loci. PMID- 2555282 TI - Effect of oxygen-derived free radicals and oxidants on the degradation in vitro of membrane phospholipids. AB - The abilities of chemically generated hydroxyl radical (OH.), superoxide anion (O.-) and hydrogen peroxide (H2O2) to degrade rat myocardial membrane phospholipids previously labeled with [1-14C]arachidonic acid were studied. HO. and H2O2, but not O2.-, caused the degradation of phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylinositol (PI). With OH. and H2O2, the loss of radiolabel in PC was accompanied by an increase in the radiolabel of lysophosphatidylcholine (LPC), but not in that of free fatty acid (FFA). These results suggest the hydrolysis of 1-oxygen ester bond of PC by HO. and that H2O2 and that HO. and H2O2, but not O.-, are detrimental to the structure and function of membrane phospholipids. However, since microM amounts of HO. and mM amounts of H2O2 were necessary to affect the membrane phospholipids, it is likely that in the reperfused myocardium only HO., but not H2O2, may directly cause the breakdown of membrane phospholipids. PMID- 2555284 TI - Intracellular oxidative cleavage of DNA in Escherichia coli by the copper-1,10 phenanthroline complex. AB - Plasmid and chromosomal DNA of E. coli during exponential growth are cleaved after treatment with copper(II)-1, 10-phenanthroline complex (1:2) without providing any exogenous reductant. About 1500 copper molecules per cell are present as estimated by atomic absorption analysis. Within the cell the endogenous reducing substances may have participated in the sequential oxidative reactions, which lead to the damage of DNA. A portion of the resultant DNA fragments originates from plasmid DNA as demonstrated by hybridization tests. PMID- 2555283 TI - Cationic polyelectrolytes: potent opsonic agents which activate the respiratory burst in leukocytes. AB - Bacteria and yeasts which are "opsonized" with cationic polyelectrolytes (poly-L arginine, poly-L-histidine and arginine-rich histone) are avidly endocytosed by both "professional" and "non-professional" phagocytes. The cationized particles also strongly activate the respiratory burst in neutrophils and in macrophages leading to the generation of chemiluminescence, superoxide and hydrogen peroxide. On the other hand, lysine and ornithine-rich polymers are poor opsonic agents. Poly L-arginine is unique in its capacity to act synergistically with lectins, with chemotactic peptides and with cytochalasin B to generate large amounts of chemiluminescence and superoxide in human neutrophils. Unlike polyarginine, polyhistidine, in the absence of carrier particles, is one of the most potent stimulators of superoxide generations, known. Neutrophils treated with cetyltrimethylammonium bromide fail to generate superoxide, but generate strong luminol-dependent chemiluminescence which is totally inhibited by sodium azide and by thiourea. Neutrophils injured by cytolytic agents (saponin, digitonin, lysolecithin) lose their chemiluminescence and superoxide-generating capacities upon stimulation by a variety of ligands. These activities are however regained by the addition of NADPH. Lysolecithin can replace polyarginine in a "cocktail" also containing lectins and cytochalasin B, which strongly activate the respiratory burst. This suggests that polyarginine acts both as a cytolytic agent and as a ligand. Arginine and histidine-rich polyelectrolytes enhance the pathogenic effects of immune complexes in vivo (reversed Arthus phenomenon) presumably by "glueing" them to tissues. Polyhistidine complexed to catalase or to superoxide dismutase, markedly enhances their efficiency as antioxidants. On the other hand polyhistidine complexed to glucose oxidase markedly enhances injury to endothelial cells suggesting that the close association of the cationized enzyme with the plasma membrane facilitates the interaction of hydrogen peroxide with the targets. A variety of cationic agents (histone, polyarginine, polyhistidine, polymyxin B) and membrane-active agents (lysophosphatides, microbial hemolysins) act synergistically with glucose oxidase or with reagent hydrogen peroxide to kill target cells. The mechanisms by which arginine- and histidine-rich polyelectrolytes activate the respiratory burst in neutrophils might involve interaction with G-proteins, the activation of arachidonic acid metabolism and phospholipase A2, or the interaction with myeloperoxidase. Naturally-occurring cationic proteins might modulate several important functions of leukocytes and the course and outcome of the inflammatory process. PMID- 2555285 TI - Effects of E. coli 0111.B4 lipopolysaccharide on spin-labelled murine macrophage and hepatocyte membranes. AB - Macrophages and hepatocytes from normal and BCG-primed mice have been spin labelled in their membranes with 5- and 16-doxyl stearic acid. Incubation of spin labelled cells from BCG-primed animals with lipopolysaccharide from E. coli 0111.B4 produced a detectable and transient disturbance in the cell membranes as reflected by an increase in the order parameter measured from the electron spin resonance spectra of 5-doxyl-stearate. This membrane disturbance was maximal at 3 4 hours of incubation and was only detected with cells from mice primed with BCG. Spectra obtained from the 16-doxyl-stearate-labelled cells showed no change in order parameter on incubation with lipopolysaccharide. PMID- 2555286 TI - Plants under drought-stress generate activated oxygen. AB - Lysed chloroplasts prepared from droughted wheat plants generate O2- on illumination as detected by electron spin resonance spectroscopy, the amplitude of the signal increasing with the severity of water deprivation. Following a similar time-course as radical formation and chlorophyll destruction, there was a significant increase in the accumulation of iron in the droughted shoots to reach an estimated concentration in the cell sap of about 2.5 mM. The evidence suggests that superoxide generated as a result of impaired electron transport in the chloroplasts reacts with the high concentration of accumulating iron resulting in the formation of hydroxyl radical, the probable cause of the primary pathologies observed in droughted plants. PMID- 2555287 TI - Ki-1 lymphoma and Hodgkin's disease. PMID- 2555289 TI - [Mucinous carcinoma of the prostate]. PMID- 2555288 TI - [Kawasaki disease associated with Epstein-Barr virus]. AB - We describe a 14-month-old child who for 3 weeks had high fever and developed lesions of the mucous membranes, enlarged lymph nodes, and an erythematous rash. Kawasaki disease was diagnosed and there was clear evidence for infection with Epstein-Barr virus. Thus it is possible that this virus is one of the etiological factors in the development of Kawasaki disease. PMID- 2555290 TI - Metabolic differentiation of bloodstream forms of Trypanosoma brucei brucei into procyclic forms. Effect of hydroxyurea, arabinosyl adenine, and serum omission. AB - Bloodforms of Trypanosoma brucei brucei STIB 247 taken from rats and containing more than 80 per cent short stumpy forms, differentiated in vitro to procyclic forms in medium SDM 79 (Brun and Schonenberger 1979), enriched with 3 mmol.dm-3 cis-aconitate. Cell division was abolished by the addition of hydroxyurea (200 micrograms.ml-1) or arabinosyl adenine (20 micrograms.ml-1 to the cultivation medium, or by the omission of serum from the medium. The ultrastructure of exponentially growing controls was rearranged within 24 h. The endogenous respiration and the respiration stimulated by proline, succinate, and 2 oxoglutarate were detectable within 12 h; after 48 h the respiration rates were comparable to those found in the established procyclic forms. After 12 h the respiration was inhibited by 200 mumol.dm-3 KCN, and by 20 mumol.dm-3 antimycin to the extent found in procyclic forms. Hydroxyurea did not significantly affect respiration. Activities of procyclic-stage enzyme markers malate dehydrogenase, threonine dehydrogenase, succinate: cytochrome c reductase, and NADH: cytochrome c reductase rose within 48 h of differentiation to values which were close to those found in established procyclic forms. The activity of glutamate dehydrogenase (NAD-specific), however, was only 1/3 of that in the procyclics, and no citrate synthase was detected in differentiating culture. Glycosomal malate dehydrogenase was detected after 6 h. In the presence of hydroxyurea or arabinosyl adenine, or in the absence of serum, respiration rates, marker enzyme activities, and glycosomal malate dehydrogenase developed to the extent comparable to the untreated controls. The results suggest that it is possible to separate the process of differentiation from cell proliferation. Cell division is not a necessary prerequisite of differentiation. PMID- 2555291 TI - [Carbonic anhydrase and its function in ion transport in cultivated pigmented ciliary body epithelial cells]. AB - The effect of inhibitors of the carbonic anhydrase on aqueous humor formation suggests that carbonic anhydrase has an important role in ion transport across the ciliary epithelium. We therefore investigated the role of carbonic anhydrase in Na+ and Cl- transport in cultured bovine pigmented ciliary epithelial cells (PE) using the radioactive isotopes 22Na and 36Cl. Our findings can be summarized as follows. (1) Na+ uptake into the cell occurs via an amiloride-sensitive Na+/H+ exchanger. (2) Cl- is transported into the cell in exchange for bicarbonate ions. (3) PE cells contain biochemical carbonic anhydrase activity. (4) Na+ uptake into NaCl-depleted cells is markedly stimulated by Cl-. This stimulation is HCO3- dependent and is completely blocked by amiloride and partly inhibited by the carbonic anhydrase inhibitor methazolamide. (5) A model is introduced for transport of NaCl in the ciliary epithelium: carbonic anhydrase is responsible for coupling of Na+/H+ exchange and Cl-/HCO3- countertransport. This model could explain the effect of carbonic anhydrase inhibitors on intraocular pressure. PMID- 2555292 TI - An improved nuclear extract preparation method. AB - A rapid, efficient, and highly reproducible procedure for nuclear extract preparation is described. The method uses lysolecithin (lysophosphatidylcholine) to disrupt plasma membranes and requires no detergents or douncing. Soluble extracts prepared by this method are comparable to conventional nuclear extracts in all assays tested. Lysolecithin nuclear extracts are competent for RNA polymerase II and III transcription, DNA replication, pre-mRNA splicing, and sequence specific DNA-protein binding. Nuclear extracts can be prepared on a small scale (10(7) cells) as well as for preparative purposes by this method. PMID- 2555293 TI - [The latent Epstein-Barr virus (EBV) activation and EBV-specific immunological abnormality in human immunodeficiency virus (HIV) infected hemophiliacs]. AB - Immunological state of HIV-infected and HIV-uninfected hemophiliacs are studied as compared with that of healthy controls. (1) EBV-specific cellular immunity observed in HIV infected hemophiliacs is significantly lower than that of HIV uninfected patients and healthy controls. (2) The titers of anti-EBNA anti-VCA antibody and anti-EA antibody in HIV-infected hemophiliacs are significantly higher than those of HIV-uninfected patients and healthy controls. Furthermore, EBV is isolated in throat washings taken from all HIV-infected hemophiliacs. (3) Surface immunoglobulin (sIg) analysis of EBV transformed lymphoblastic cell lines (LCL) revealed that sIgD positive cell percentage in HIV-infected patients is significantly higher than those of HIV-uninfected patients and healthy controls. Based on these results, it is suggested that HIV infection may give rise to the reactivation of latent EBV and may relate in not only the defect of severe cellular immunity but also the derangement of humoral immunity. PMID- 2555294 TI - Effect of heat-stable factor (SLF3) on plasma and erythrocyte cholesterol levels and erythrocyte (Na+,K+)-ATPase activity in cholesterol fed rabbits. PMID- 2555295 TI - Does hypoprolactinaemia affect adrenal androgen levels in children with hypopituitarism? PMID- 2555296 TI - Tumor necrosis factor-alpha induces a kappa B sequence-specific DNA-binding protein in human hepatoblastoma HepG2 cells. AB - Tumor necrosis factor-alpha is an inducer of acute-phase protein synthesis in liver cells. The mechanism by which tumor necrosis factor-alpha alters gene expression in these cells is largely unknown. In this study, we demonstrate that tumor necrosis factor-alpha stimulates human immunodeficiency virus-1 long terminal repeat-promoted gene expression in the human hepatoblastoma HepG2 cell line and increased binding of trans-activating factors to kappa B (kappa B) DNA sequences. In contrast to lymphocytic cells where the nuclear factors recognizing the kappa B sequences are activated by both tumor necrosis factor-alpha and phorbol-12-myristate-13-acetate through a posttranslational mechanism, in HepG2 cells phorbol-12-myristate-13-acetate does not activate these factor(s), and de novo protein synthesis seems to be required in HepG2 cells for gene activation by tumor necrosis factor-alpha. PMID- 2555297 TI - Monitoring of sodium:proton exchange in isolated hepatocytes by electronic cell sizing. AB - To investigate volume-regulating processes in the hepatocyte, a rapid and precise method of measuring cell volume in isolated hepatocytes was devised which uses a Coulter Counter equipped with a Channelyzer. Isolated hepatocytes exhibit a volume-decreasing mechanism (potassium channel) which is triggered by cell volume increases as small as 10%. Cell volume increases in the hepatocyte may be mediated by activity of the Na:H exchanger. To examine Na:H exchange-mediated cell volume increases, without apparent interference by the volume-decreasing mechanism, acetate was substituted for chloride in the incubation medium. Hepatocytes placed in a medium containing sodium acetate at an acidic pH exhibit a continuous amiloride-sensitive swelling. A simple procedure was devised for estimating Na:H exchanger set-point by electronic cell sizing. In a sodium acetate medium, the internal pH equilibrates with the external pH. By placing cells in sodium acetate medium of various pH values and measuring the rate of amiloride-sensitive swelling, an estimate of Na:H exchanger set-point can be obtained. By this method, the exchanger was estimated to cease activity above an intracellular pH of 7.2. This method could be useful for identification of stimuli that might promote cell enlargement by raising the exchanger set-point. The phorbol ester 12-O-tetradecanoylphorbol-13-acetate tetraacetic acid raises the set-point of the exchanger in the isolated hepatocytes, resulting in exchanger activity at normal cellular pH, and at the same time promotes hepatocyte swelling. Exchanger activation via a kinase C-mediated mechanism is one possible way that hepatocyte enlargement may occur. PMID- 2555299 TI - Nonspecific immunoprotection against Rous sarcoma tumor in chicks. AB - An enzyme treated preparation of saprophytic Mycobacterium phlei, referred as NSI, when administered intramuscularly has been found to protect the chicks against Rous Sarcoma Virus induced tumor. A protection level of 35.4%, 24.1% and 21.2% were observed when challenged on 10th, 20th and 30th day post NSI inoculation. The tumor growth inhibitory-activity of NSI was significant (P less than 0.01). Both, systemic and intralesional administration of NSI exhibited significant tumorostatic activity (P less than 0.05). NSI stimulated the cell mediated immune response to specific as well as to nonspecific Rous sarcoma antigen. These studies indicated the immunopreventive activity of NSI against Rous sarcoma tumor which had an immunogenic basis. PMID- 2555298 TI - Prevalence of hepatocellular carcinoma and relation to cirrhosis: comparison of two different cities of the world--Trieste, Italy, and Chiba, Japan. AB - A comparative, descriptive study on the prevalence of hepatocellular carcinoma either associated or not with cirrhosis was undertaken between the cities of Trieste, Italy, and Chiba, Japan. Data were derived from 12,340 and 2,052 autopsies consecutively performed in Trieste and Chiba during 1980 to 1984 and 1975 to 1985, respectively. During these years, more than 70% of all deaths were studied by autopsy in Trieste. Clinical cases of hepatocellular carcinoma admitted to the Chiba University Hospital were also analyzed to correct for the bias induced by the relatively small number of autopsies in Chiba. Serum hepatitis B surface antigen was more frequent in Chiba, whereas alcohol intake was greater in Trieste. From the comparison of the data, we conclude: (i) hepatocellular carcinoma was associated with cirrhosis in the large majority of the cases both in Trieste (87%) and Chiba (81%); (ii) in both sexes, the average age of hepatocellular carcinoma patients was significantly older (p less than 0.005) in Trieste than in Chiba; (iii) in the autopsy series, cirrhosis was more frequent in Trieste (9.7%) than in Chiba (1.5%); (iv) in Chiba, hepatocellular carcinoma was more frequent than cirrhosis not complicated by hepatocellular carcinoma (6.9 vs. 1.5%) at necroscopy, and (v) the age-standardized rate of hepatocellular carcinoma was significantly (p less than 0.01) higher in Trieste than in Chiba. These geographical differences might be related to the different type of cirrhosis observed in Trieste (alcoholic) and Chiba (postnecrotic). PMID- 2555300 TI - Pattern of enzyme changes in rabbits administered linamarin or potassium cyanide. AB - Diseases like tropical ataxic neuropathy and endemic goitre have been reported to have definite correlation with a chronic ingestion of cassava (Manihot esculenta Crantz). The toxicity of cassava has been attributed to its two cyanogenic glycosides, linamarin and lotaustralin. In this study, an attempt has been made to understand the pattern of changes in certain clinically significant enzymes brought about by the chronic administration of sublethal doses of linamarin to rabbits. The profound elevation in rhodanese activity observed in the linamarin and cyanide treated rabbits indicated the attempt of the tissues to detoxify cyanide. That intact linamarin could be hydrolysed in vivo was a significant finding from the study. The mode of toxicity of linamarin was similar to that of cyanide by producing a gradual shift from aerobic to anaerobic metabolism. PMID- 2555301 TI - Racial differences in ion regulation and their possible links to hypertension in blacks. PMID- 2555302 TI - Ion transport defects and hypertension. Where is the link? PMID- 2555303 TI - Altered pituitary hormone response to naloxone in hypertension development. AB - Endogenous opioid regulation of blood pressure is altered during stress in young adults at risk for hypertension. We studied the effects of the opioid antagonist naloxone on the secretion of corticotropin and beta-endorphin during psychological stress in young adults with mildly elevated casual arterial pressures. Naloxone-induced secretion of both corticotropin and beta-endorphin was significantly diminished in persons at enhanced risk for hypertension compared with the low blood pressure control group. Results suggest that opioidergic inhibition of anterior pituitary function is altered in hypertension development. PMID- 2555304 TI - Atrial natriuretic factor modulates proximal glomerulotubular balance in anesthetized rats. AB - The extent to which the natriuretic effect of a prolonged low dose infusion of atrial natriuretic factor (30 ng/kg/min) is dependent on interference with the prevailing intrarenal actions of angiotensin II was examined before and after blockade of angiotensin production with the converting enzyme inhibitor enalaprilat (5 mg/kg). Lithium clearance was used to assess proximal tubular sodium and water reabsorption. Atrial natriuretic factor and enalaprilat caused similar increases in sodium excretion (10-fold and sevenfold, respectively) and glomerular filtration rate (each 34%) and similar decreases in fractional proximal reabsorption of sodium (17% and 13%, respectively) and blood pressure. Each also caused a major disruption in the effectiveness of proximal glomerulotubular balance (30% and 50% of perfect balance). Infusion of atrial natriuretic factor during converting enzyme inhibition increased glomerular filtration rate further by 23%, reaching 63% above control without change in renal blood flow but with a rise in filtration fraction to 0.48. Sodium excretion increased further but fractional proximal sodium reabsorption remained constant and proximal glomerulotubular balance appeared to improve. Atrial natriuretic factor therefore possesses a glomerular action that persists during converting enzyme inhibition and is indeed additive to the removal of angiotensin II when the proximal effect of atrial natriuretic factor is no longer apparent. It is concluded that failure of atrial natriuretic factor to further suppress fractional proximal sodium reabsorption during converting enzyme inhibition is caused by either prior removal of the stimulatory action of angiotensin II on proximal tubular transport or extreme changes in peritubular physical factors consequent on the high filtration fraction. PMID- 2555305 TI - Subarachnoid hemorrhage--an unusual presentation of cerebral tumour in children. PMID- 2555306 TI - Cyclic AMP inhibits chemotactic-peptide-induced but not Ca2+-ionophore- or tetradecanoylphorbol-acetate-induced enzyme secretion in guinea pig neutrophils. AB - cAMP-increasing agents such as prostaglandin El, dibutyryl cAMP and 8-bromo-cAMP inhibited N-acetyl-beta-D-glucosaminidase secretion induced by the chemotactic peptide formylmethionyl-leucyl-phenylalanine in guinea pig neutrophils. However, they could not inhibit but rather potentiated the secretion significantly when secretion was induced by the Ca2+ ionophore A23187. Similarly, these agents did not inhibit tetradecanoylphorbol acetate (TPA)-induced secretion. Moreover, the secretion induced by combined treatment with Ca2+ ionophore A23187 and TPA was hardly blocked by cAMP-increasing agents. These results strongly suggest that cAMP inhibits the secretion through suppression of a signal transduction from receptor activation to events such as Ca2+ mobilization and protein kinase C activation, probably of the inositol phospholipid metabolism. PMID- 2555307 TI - Activation and immortalization of leukaemic B cells by Epstein-Barr virus. AB - We have studied the responses of chronic leukaemic B cells to infection by Epstein-Barr virus (EBV). Our results define one population of B lymphocytes, represented by prolymphocytic leukaemic (PLL) cells, which are highly susceptible to immortalization by EBV. Another B-cell type, represented by chronic lymphocytic leukaemic (CLL) cells, can be readily infected by the virus but is resistant to immortalization. Comparative studies of viral and cellular related events early after infection in these 2 cell types reveal that both express the EB viral nuclear antigen (EBNA) complex, but the immortalization-resistant CLL cells fail to express the latent membrane protein (LMP), which can be detected in PLL cells 48 hr after infection. Circularization of the linear viral genome could be detected at 7 days post infection in the PLL cells, but only in 2 out of 4 CLL cells tested. Both CLL and PLL cells show increased surface expression of CD23 and HLA-DR molecules after infection but, whereas PLL cells show an increase in size, together with RNA and DNA synthesis indicative of cell cycle progression, CLL cells appear to be arrested in the G1/S phase of the cycle. The results suggest that the outcome of infection by EBV is determined by the nature of the target cell rather than by random virus-related events. The correlation between the block in immortalization of CLL cells and their failure to express LMP suggests that expression of this protein is essential for in vitro immortalization of B cells. The failure to detect circularization in some EBV infected CLL cells suggests that this, as well as LMP expression, may be dependent on prior activation of the B cell by EBV, an event which may vary between the different CLL samples tested. PMID- 2555308 TI - Suppression of focus formation by bovine papillomavirus-transformed cells by contact with non-transformed cells: involvement of sugar(s) and phosphorylation. AB - Focus formation by bovine papilloma virus-transformed C127 cells was inhibited by direct contact with non-transformed C127 cells. The suppressive capacity of C127 cells was abolished by introduction of the neomycin resistance gene (neor) but not by that of the hygromycin resistance gene (hygrr). Though both genes code for phosphotransferase which inactivates the aminoglycoside antibiotics, their substrates are different, i.e., there is no cross-resistance between them. As the neomycin phosphotransferase phosphorylates the specific hydroxyl group of the sugar in the aminoglycosides, such as 3'OH of the glucose residue of kanamycin A, some specific sugar(s) on the molecules exposed on the cell surface must be responsible for the suppressive signal and their phosphorylation must have resulted in the loss of that signal. The sugar must have the structure shared by kanamycin, neomycin or G418 but not by hygromycin B. Involvement of sugar was also suggested by the observation that concanavalin A partially abrogated the suppressive capacity of C127 cells. PMID- 2555309 TI - Clustering of discrete cell properties essential for tumorigenicity and metastasis. II. Studies of Syrian hamster embryo fibroblasts transformed by Rous sarcoma virus. AB - Tumorigenic and metastasizing activities (TGA; MA) and susceptibility, or resistance to H2O2 and PGE-releasing activity (H2O2R + PGEs+ phenotype) have been examined in 6 Syrian hamster embryo cell strains transformed in vitro with Rous sarcoma viruses (Schmidt-Ruppin and Prague strains). Early observations of extremely high level of TGA and even MA of RSV-SR-transformants never selected in vivo have been confirmed. The correspondence of these properties with a high level of expression of H2O2R + PGEs+ phenotype and its clustering character were demonstrated in 4 RSV-SR transformants, while significantly lower expression of all these characteristics, including TGA, was observed in 2 RSV-Prague transformants. High level of spontaneous MA was noticed in some RSV-SR transformants. A tumor cell line induced in vivo by RSV-SR did not differ from the cell strain transformed in vitro by RSV-SR. Inhibition of H2O2R + PGEs+ phenotype in one of RSV-SR transformants was obtained with non-toxic doses of BCNU and indomethacin, leading to a marked decrease of TGA. PMID- 2555310 TI - Diacylglycerols and the protein kinase inhibitor H-7 suppress cell polarity and locomotion of Walker 256 carcinosarcoma cells. AB - We show that diacylglycerols, like phorbol myristate acetate (PMA), suppress cell polarity and locomotor activity of Walker carcinosarcoma cells in a dose dependent fashion in vitro. OAG and diC8 show significant activity at concentrations above 3 x 10(-5) M. The inhibitory effect on locomotion is due to a reduction in the proportion of locomoting cells rather than gradual lowering of the speed of individual cells. Measurement of protein kinase C (PKC) activity in isolated fractions showed a substantial reduction of the total cellular PKC activity and of the activity in the cytosolic fraction following incubation of cells with 10(-8) M PMA for 30 min. In contrast, the total and relative PKC activity associated with the membrane fraction was increased by PMA. The effect of H-7, an inhibitor of PKC as well as of cAMP-dependent kinase, has been tested. H-7 suppressed cell polarity of "unstimulated" control cells (ID50 = 6.5 microM H 7), colchicine-stimulated cells (ID50 = 92 microM H-7) or cells treated with both PMA and colchicine (ID50 = 15 microM H-7), in a dose-dependent fashion. The locomotor activity of the cells was also suppressed. LTB4 had no clearcut activity in this system. Our findings suggest that diacylglycerols and H-7 are of interest as physiological or pharmacological stop signals for tumor-cell locomotion. Contrary to our expectations, PMA and diacylglycerols vs. H-7 did not produce opposing or antagonistic effects on cell polarity and locomotion. This similarity may be due to down-regulation of PKC by PMA and inhibition of PKC by H 7. However, the mechanisms underlying these novel effects of diacylglycerols and of H-7 on cell polarity and locomotion may be even more complex; they require further studies. PMID- 2555312 TI - Giant cell tumor of tendon sheath. AB - The giant cell tumor of tendon sheath (localized nodular tenosynovitis) is the second most common tumor involving the hand but is only rarely reported in the dermatologic literature. A case of giant cell tumor and a review of the clinical and pathologic records of 111 patients are discussed. PMID- 2555311 TI - The bioavailability of sulpiride taken as a film-coated tablet with sodium bicarbonate, cimetidine, natural orange juice or hydrochloric acid. AB - The bioavailability of sulpiride taken in film-coated tablet form with sodium bicarbonate or cimetidine or with natural orange juice or diluted hydrochloric acid was studied. A commercial sulpiride film-coated tablet (100 mg/T) treated with polyvinylacetal diethylaminoacetate (AEA), which remain undissolved at pH above 4-5, was given to four healthy volunteers who had fasted overnight. The subjects were divided into two groups, those showing high and low bioavailability of sulpiride from an AEA film-coated tablet. The two high bioavailability subjects took one tablet (100 mg) with 100 ml of water (1) alone, (2) together with 1 g of sodium bicarbonate or (3) during concurrent dosing with cimetidine, 200 mg three times a day. The two low bioavailability subjects swallowed one tablet with 100 ml of (1) water, (2) natural orange juice, or (3) diluted hydrochloric acid. Urine samples were collected over a 48-h period following sulpiride administration to determine sulpiride concentrations by HPLC. The bioavailability was estimated from the cumulative amount excreted unchanged in urine over 48 h (Du48). In the high bioavailability subjects, the bioavailability of sulpiride markedly decreased with the coadministration of sodium bicarbonate or cimetidine compared to when the tablet was taken alone. In the low bioavailability subjects, the bioavailability remarkably increased with the concomitant intake of orange juice or diluted hydrochloric acid over that with only water. These results suggest that the bioavailability of sulpiride from AEA film-coated tablet is influenced by the individual's gastric acidity and by coadministered drug and drink which affect gastric acidity. PMID- 2555313 TI - Calcium-dependent phosphodiesterase from male mouse germ cells: functional properties and characterization of the enzyme. PMID- 2555314 TI - Intraoral adenoid cystic carcinoma: the role of postoperative radiotherapy in local control. AB - Fourteen cases of adenoid cystic carcinoma (ACC) of the intraoral salivary glands with positive surgical margins have been reviewed in order to determine the role of postoperative radiotherapy in local control. Since local control was obtained in all patients, postoperative radiotherapy seems an adequate treatment to deal with the problem of positive surgical margins at the microscopic level, in cases of intraoral ACC, making additional surgical treatment redundant. PMID- 2555315 TI - Destruction of genotoxic wastes mixed with radioactive products. AB - Before their disposal, genotoxic substances are destroyed by strong oxidizing agents. If there are molecules labelled with radionuclides in the medium which is oxidized, then this treatment may bring about the release of gaseous radioactive compounds. We have looked for evidence of such a release following the action of K permanganate and sodium hypochlorite on molecules labelled with 3H, 14C, 32P, and 125I. Among the compounds examined, only those labelled with 14C showed significant quantities of radioactive gas released, with values up to 60% of the total radioactivity. For the other products, less than 0.6% of the radioactivity appeared in a volatile form. PMID- 2555316 TI - Measurements of radioactive gaseous releases to air from target halls at a high energy proton accelerator. AB - Measurements of induced radioactivity concentration in air were made at five target halls associated with the Fermilab high-energy proton synchrotron. A gas flow-through ionization chamber was used to continuously sample air released from an exhaust stack at each location. The radioactive decay of a grab sample of air from each target hall was recorded, and a best-fit of each decay curve was made via iterative steps to determine the mixture of radionuclides present. The radionuclides 11C, 13N, 15O, 38Cl, 39Cl, and 41Ar were identified. Two distinct radionuclide mixtures or "signatures" were found and were correlated to the geometry of the proton loss point. The normalized activity release rates for the five target areas agree within a factor of two of the average value of 2.7 muBq per proton for 800 GeV protons. PMID- 2555317 TI - Were hospitals selective in their product and productivity changes? The top 50 DRGs after PPS. AB - Five separate hospital products are identified based on the concept of the amount of disease remission achieved by the hospital. The parameters of this concept are illness level on admission and discharge location. In a cohort of 646 nonfederal, short-term hospitals over the period 1980-1984, changes in the hospital product are examined separately in the 50 diagnosis-related groups (DRGs) with the greatest volume of Medicare discharges. Productivity changes, as defined by the number of certain inputs, are also examined. In both sets of analyses, patient severity level is controlled for by indexing to the base year (1980) case mix. The purpose of this study was to examine whether the dramatic product and productivity changes following implementation of the prospective payment system, as found in our earlier work, were across-the-board changes or the result of selective changes, specific to certain DRGs or products. The results suggest that the changes were an across-the-board phenomenon. Policy implications are discussed. PMID- 2555318 TI - Collagenase activity in the normal rat myocardium. An immunohistochemical method. AB - Fibrillar collagen in the myocardium provides a supportive framework for myocytes and capillaries. Disruption of this organized framework has been observed in certain pathological states. Collagen degradation is primarily mediated by the specific enzyme collagenase, which has been found to exist in various tissues including the myocardium. In this report we describe a method that detects collagenase activity in sections of cardiac tissue. This method is on the basis of degradation of collagen by collagenase on one hand and the visualization of disrupted collagen fibers by immunofluorescence on the other. Frozen rat heart sections were incubated under optimal conditions for collagenase activity (37 degrees C in the presence of 0.1 M calcium at pH 7.4) for 24 h and 48 h. Subsequently, immunofluorescence staining with antibody to type I collagen was performed and the collagenous structures were visualized by immunofluorescence light microscopy. As control, untreated rat heart sections and sections incubated in the absence of calcium were similarly treated with antibody. After the 24 h of incubation, we found no change in the structural integrity of collagen fibers. Marked disruption of the type I collagen fibers was observed 48 h after incubation. No evidence of collagen fiber disruption was found in control sections. Experiments with exogenous collagenase resulted in similar collagen fiber disruption in the frozen rat heart sections. We conclude that the disruption of collagen type I fibers after 48 h of incubation, under optimal conditions for collagenolytic digestion, is the result of collagen degradation by intrinsic collagenase of the myocardium. PMID- 2555319 TI - Inhibition of in vitro tumor cell invasion by transmethylation inhibitors. AB - Three inhibitors of S-adenosylmethionine-mediated transmethylation, 5' methylthioadenosine (MTA), 2'-deoxyadenosine and sinefungin, inhibited in vitro invasion by a highly invasive clone (Cl-30) of rat ascites hepatoma cells, AH 130 (AH cells). Difluoromethylthioadenosine (DFMTA), a non-metabolizable derivative of MTA, also caused strong inhibition of invasion at concentrations that did not suppress the growth of the tumor cells. Cl-30 cells precultured in methionine depleted medium showed decreased invasiveness. DFMTA was also effective on the invasion by fibrosarcoma, B16 melanoma and human lung carcinoma cell lines. PMID- 2555321 TI - Enzymes involved in the biosynthesis of leukotriene B4. PMID- 2555320 TI - Serial measurement of sister chromatid exchanges in the peripheral lymphocytes of patients with lung cancer receiving chemotherapy in relation to bone marrow toxicity. AB - The frequencies of sister chromatid exchanges (SCE) in the peripheral lymphocytes of patients with lung cancer before and after initial chemotherapy were serially measured and the correlation between SCE frequencies and bone marrow toxicity was evaluated. The addition of mitomycin C to vindesine plus cisplatin increased SCE frequencies significantly in patients with non-small cell lung cancer. A significant increase in frequencies of SCE was observed in patients treated with cyclophosphamide plus adriamycin plus vincristine as compared with those treated with cisplatin plus etoposide in small cell lung cancer. A significant inverse correlation was observed between SCE frequencies in the peripheral lymphocytes 7 days after treatment (x) and the nadir value/pretreatment value of platelets (y)(r = -0.685, P = 0.0007, y = 0.842-0.022x). The relation between SCE frequency and leukopenia showed the same trend as thrombocytopenia, but the correlation was not statistically significant (r = -0.444, P = 0.057). SCE assay may have potential clinical use for the prediction of chemotherapy-induced thrombocytopenia. PMID- 2555322 TI - Agonist-induced desensitization of a P2Y-purinergic receptor-regulated phospholipase C. AB - A guanine nucleotide-dependent P2Y-purinergic receptor-regulated phospholipase C activity of turkey erythrocyte membranes has been characterized in detail previously (Boyer, J. L., Downes, C. P., and Harden, T. K. (1989) J. Biol. Chem. 264, 884-890). The occurrence of agonist-induced desensitization of this receptor regulated phospholipase C is now described. Preincubation of turkey erythrocytes with the P2Y-purinergic receptor agonist ADP beta S resulted in a marked loss of capacity of ADP beta S plus GTP to stimulate phospholipase C in membranes derived from these cells. The half-time of occurrence of desensitization was 0.5-2.0 min, and within 10 min responsiveness had reached a new quasi-steady state level representing 40-55% of control. Transfer of agonist-preincubated erythrocytes to agonist-free medium resulted in recovery of agonist plus GTP responsiveness of the membrane phospholipase C activity to control levels with a half-time of 10-20 min. The change in ADP beta S plus GTP responsiveness occurred as a loss of maximal effect with little or no change in the apparent affinity of agonist for stimulation of inositol phosphate production. Induction of desensitization occurred with an agonist-specificity that followed that expected of a P2Y purinergic receptor. Neither the rate of activation nor the final phospholipase C activity attained in the presence of GTP gamma S alone was altered in membranes from cells preincubated with ADP beta S for 15 min. AlF-4-stimulated inositol phosphate production was also not modified in membranes from agonist-preincubated erythrocytes. In contrast, the capacity of ADP beta S to increase the rate of activation of phospholipase C by GTP gamma S was markedly reduced in membranes from agonist-preincubated cells. The amount of 3H-radioactivity in phosphoinositides, as well as the ratio of labeling among the phosphoinositides, was not altered by incubation of erythrocytes with a P2Y-purinergic receptor agonist. Taken together these data suggest that P2Y-purinergic receptor agonist induced desensitization occurs as a consequence of a modification at the level of the receptor or at the level of receptor-guanine nucleotide regulatory protein (G protein) coupling with no change occurring in the capacity of the G-protein to activate phospholipase C. PMID- 2555323 TI - Thrombin induces a calcium transient that mediates an activation of the Na+/H+ exchanger in human fibroblasts. AB - The calcium dependence of growth factor-induced cytoplasmic alkalinization was determined in serum-deprived human fibroblasts (WS-1 cells). Intracellular pH (pHi) and intracellular calcium (Ca2+i) were measured using the fluorescent dyes 2',7'-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein and fura2, respectively. Thrombin (10 nM) induced an alkalinization (0.18 +/- 0.01 pH units, n = 23) that was Na+-dependent and amiloride-sensitive, suggesting that the alkalinization was mediated by the Na+/H+ exchanger. Thrombin treatment caused a transient increase in Ca2+i (325 +/- 39 nM, n = 12) that preceded the observed increase in pHi. The increases in Ca2+i and pHi were dependent on the concentration of thrombin. The thrombin-induced increase in Ca2+i occurred in the absence of external calcium indicating that thrombin released calcium from internal stores. Inhibition of the thrombin-induced increase in Ca2+i with 8-diethylaminooctyl 3,4,5 trimethoxybenzoate hydrochloride or bis-(o-aminophenoxy)ethane-N,N,N',N'- tetraacetic acid also inhibited the thrombin-stimulated increase in pHi. The calcium ionophore ionomycin was used to increase Ca2+i independent of growth factor stimulation. When Ca2+i was elevated with ionomycin, a concomitant increase in pHi was observed. The increase in pHi due to ionomycin was dependent on Na+ and sensitive to amiloride. The removal of external Ca2+i inhibited the ionomycin-induced elevation of both Ca2+i and pHi. The ionomycin-induced increases in Ca2+i and pHi were not inhibited by 8-diethylaminooctyl 3,4,5 trimethoxy-benzoate hydrochloride. The results suggest that thrombin treatment can activate the Na+/H+ exchanger, and this activation is mediated by an increase in Ca2+i. PMID- 2555324 TI - A model for the stepwise radiation inactivation of the alpha 2-dimer of Na,K ATPase. AB - This study is a direct continuation of Jensen, J., and Norby, J. G., (1988) J. Biol. Chem. 263, 18063-18070. A new model in which we propose that the in situ organization of the Na,K-ATPase alpha-subunit is an alpha 2-dimer and which describes the stepwise degradation by radiation inactivation of this assembly is presented on the basis of the following findings. Radiation inactivation size for alpha-peptide integrity, normal nucleotide, vanadate and ouabain binding, and K pNPPase activity is close to m(alpha) = 112 kDa; for Na-ATPase activity it is 135 kDa and for Na,K-ATPase activity it increases from 140 to about 195 kDa with increasing assay ATP concentration (equal to increasing average turnover). Normal Tl+ occlusion had the same radiation inactivation size as Vmax for Na,K-ATPase, i.e. about 195 kDa. The binding experiments disclosed radiation-produced molecules with active binding sites but with a lower than normal affinity. Radiation inactivation size for the total binding capacity of ADP and ouabain was therefore smaller than the size of an alpha-peptide, namely about 70 kDa, and for total Tl+ occlusion it was down to 40 kDa. We can explain all these observations by using a new approach to target size analysis and by assuming a dimeric organization of the alpha-subunit. Each alpha-peptide is degraded stepwise by first destruction of either a 42- or a 70-kDa domain, and the partly damaged peptide may retain biochemical activity. We conclude that there is no role for the beta-subunit in catalysis and that the alpha-peptide is organized as an alpha 2-dimer in the membrane with each alpha-subunit being able to perform complete catalytic cycles (and probably also active transport), provided that it is stabilized by an adjacent alpha-peptide or a sufficiently large fragment thereof. PMID- 2555325 TI - Cleavage specificity of type IV collagenase (gelatinase) from human skin. Use of synthetic peptides as model substrates. AB - Type IV collagenase (gelatinase) has a marked substrate specificity for denatured collagen (gelatin). Cleavage site specificity of type IV collagenase from human skin was determined using small collagenous peptides with varied sequences around Gly-Leu or Gly-Ile. Type IV collagenase showed essentially the same order of preference for the peptide substrates as did interstitial collagenase. Both required a peptide with a minimum of six amino acid residues to demonstrate significant gelatinolytic activity and were able to cleave uncharged molecules more rapidly than charged molecules. the repeating Gly-X-Y-Gly sequence of collagen is not an absolute requirement for either enzyme since both digested AcPro-Leu-Gly-Ile-Leu-Ala-Ala-OC2H5 at 70% of the rate of the best substrate peptide, AcPro-Leu-Gly-Leu-Leu-Gly-OC2H5. Km and kcat (Vmax) values were determined for several of the peptides and for the native substrate. Turnover numbers with type IV collagenase were similar to those with interstitial collagenase (Weingarten, H., Martin, R., and Feder, J. (1985) Biochemistry 24, 6730-6734). However, the Km for all peptides investigated was approximately 10 fold lower for type IV collagenase than for interstitial collagenase. Because type IV collagenase does not cleave helical interstitial collagens, the data support the conclusion that secondary structure determines whether the peptide bond can be hydrolyzed at any potential cleavage site. PMID- 2555326 TI - Evidence for the activation of two different Ca2+ channels during the egg jelly induced acrosome reaction of sea urchin sperm. AB - The influx of Ca2+ and its subsequent intracellular increase are required for the acrosome reaction of sea urchin sperm to occur. Spermatozoa must undergo this reaction, which is triggered by the egg jelly, in order to fertilize the egg. Here, the egg jelly-induced Ca2+ influx mechanisms have been studied in sperm loaded with FURA-2 using Mn2+ under the assumption that this divalent ion is an indicator of Ca2+ influx through Ca2+ channels. Egg jelly induced the immediate entry of Ca2+ (mixing time 1 s), however; we found that the influx of Mn2+ increased after a lag time of 5 s. Nisol-dipine (a Ca2+ channel blocker) did not block the Mn2+ influx which was inhibited by 40 mM of external [K+], low Na+, and 5 mM of tetraethylammonium (a K+ channel blocker). These conditions also inhibited the alkalinization and the acrosome reaction. The inhibition of the Mn2+ influx could be overcome by increasing internal pH (pHi) with ammonium (10 mM). On the contrary the influx of Ca2+ during the first 5 s was not inhibited by any of the conditions indicated before, except by nisoldipine. These data could be explained by the activation of two different Ca2+ channels by egg jelly. The first one being a receptor-operator Ca2+ channel that opens when the receptor for egg jelly is occupied independently of the ionic conditions. The other one could be considered as a second messenger-operated Ca2+ channel that requires at least an increase in pHi to open. PMID- 2555327 TI - Tryptic fragments of the Escherichia coli DNA gyrase A protein. AB - Treatment of the Escherichia coli DNA gyrase A protein with trypsin generates two large fragments which are stable to further digestion. The molecular masses of these fragments are 64 and 33 kDa, and they are shown to be derived from the N terminus and the C terminus of the A protein, respectively. These fragments could represent structural and/or functional domains within the A subunit of DNA gyrase. The trypsin-cleaved A protein (A'), in combination with the B subunit of gyrase, can support ATP-dependent supercoiling of relaxed DNA and other reactions of DNA gyrase. The isolated 64-kDa fragment will also catalyse DNA supercoiling in the presence of the B protein, but the 33-kDa fragment shows no enzymic activities. We conclude that the N-terminal 64-kDa fragment represents the DNA breakage/reunion domain of the A protein, while the 33-kDa fragment may contribute to the stability of the gyrase-DNA complex. PMID- 2555328 TI - Purification and properties of the phosphotriesterase from Pseudomonas diminuta. AB - The phosphotriesterase produced from the opd cistron of Pseudomonas diminuta was purified 1500-fold to homogeneity using a combination of gel filtration, ion exchange, hydrophobic, and dye matrix chromatographic steps. This is the first organophosphate triesterase or organophosphofluoridate hydrolyzing enzyme to be purified to homogeneity. The enzyme is a monomeric, spherical protein having a molecular weight of 39,000. A single zinc atom is bound to the enzyme and is required for catalytic activity. Incubation with metal chelating compounds, o phenanthroline, EDTA, or 2,6-pyridine dicarboxylate inactivate the enzyme. The kinetic rate constants, kcat and kcat/Km, for the hydrolysis of paraoxon are 2100 s-1 and 4 x 10(7) M-1 s-1, respectively. The enzyme is inhibited competitively by dithiothreitol, dithioerithritol, and beta-mercaptoethanol. In addition to paraoxon the phosphotriesterase was found to hydrolyze the commonly used organophosphorus insecticides, dursban, parathion, coumaphos, diazinon, fensulfothion, methyl parathion, and cyanophos. PMID- 2555329 TI - Regulation by high density lipoproteins of muscarinic acetylcholine receptor function in chick heart cells cultured in defined medium. AB - Activation of cardiac muscarinic acetylcholine receptors (mAChR) on cultured chick heart cells results in a decrease in cellular cAMP levels and a stimulation of phosphoinositide breakdown. A serum-free culture system has been used to investigate the regulation of mAChR number and function by purified serum high density lipoprotein (HDL). Administration of HDL purified from rooster serum to chick heart cells cultured in defined medium results in an attenuation of the ability of muscarinic agonist to inhibit forskolin-stimulated cAMP accumulation, with no change in its ability to stimulate phosphoinositide hydrolysis or to mediate down-regulation of receptor number. The inclusion of HDL in the culture medium did not result in appreciable changes in mAChR number or affinity, nor were the levels of the inhibitory guanine nucleotide-binding regulatory proteins (G-proteins) altered. However, the ability of guanine nucleotides to inhibit forskolin-stimulated adenylate cyclase activity was reduced by HDL treatment, suggesting that HDL interferes with the capacity of G-proteins to interact with adenylate cyclase. In order to determine which component of native HDL mediates the decreased effectiveness of carbachol, the ability of lipid and apoprotein fractions to mimic the effect of HDL was tested. HDL lipid fractions were able to mimic the effect of native HDL, while protein fractions were not. This result suggests that the ability of HDL to attenuate muscarinic receptor function is mediated by its lipid constituents. The effect of HDL and HDL lipid fractions were not correlated with changes in membrane cholesterol content. PMID- 2555330 TI - Oxygen-based free radical generation by ferrous ions and deferoxamine. AB - Deferoxamine accelerates the autooxidation of iron as measured by the rapid disappearance of Fe2+, the associated appearance of Fe3+, and the uptake of oxygen. Protons are released in the reaction. The formation of H2O2 was detected by the horseradish peroxidase-catalyzed oxidation of scopoletin, and the formation of hydroxyl radicals (OH.) was suggested by the formation of the OH. spin trap adduct (DMPO/OH). with the spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) and the generation of the methyl radical adduct on the further addition of dimethyl sulfoxide. (DMPO/OH). adduct formation was inhibited by catalase but not by superoxide dismutase. The oxidant formed converted iodide to a trichloroacetic acid-precipitable form (iodination) and was bactericidal to logarithmic phase Escherichia coli. Both iodination and bactericidal activity was inhibited by catalase and by OH. scavengers, but not by superoxide dismutase. Iodination was optimal in 5 x 10(-4) M acetate buffer, pH 5.0, and when the Fe2+ and deferoxamine concentrations were equimolar at 10(-4) M. Fe2+ could not be replaced by Fe3+, Co2+, Zn2+, Ca2+, Mg2+, or Mn2+, or deferoxamine by EDTA, diethylenetriaminepentaacetic acid, or bathophenanthroline. These findings indicate that Fe2+ and deferoxamine can act as an oxygen radical generating system, which may contribute to its biological effects in vitro and in vivo. PMID- 2555331 TI - Transient response of retinal rod outer segment phosphodiesterase to actinic light pulses. I. Simple quantitative kinetic model. AB - We present a quantitative kinetic model for the transient velocity (microM of cGMP hydrolyzed/s) response of retinal rod outer segment (ROS) cGMP phosphodiesterase (v(t) versus t) to a stimulating light pulse in the linear response range. The model gives an excellent fit to experimental v(t) versus t data for ROS suspensions at different concentrations of GTP and GDP and clarifies experimental results which are difficult to understand in the absence of such a model. It contains the minimum number of steps required to fit our experimental data and consists of one rate-limiting step with specific rate kL for the production of active phosphodiesterase (PDE), PDE*, by photoactivated rhodopsin, R*, and deactivation processes for R* and PDE* with lifetimes tau R and tau P, respectively. The experimental graphs of v(t) versus t at each concentration of GTP and GDP are characterized by a fast rise to a peak value, vpeak, followed by a slow decay to zero level. The minimal kinetic model allows us to characterized completely the effects of GTP and GDP, and any other pertinent species, in terms of their effects on the parameters kL, tau R, and tau P. Our kinetic model indicates that for "washed" ROS preparations (a) the risetime of v(t) is determined by tau P which has a value of about 2 s and is insensitive to [GTP]. (b) The decay of v(t) is determined by tau R which decreases with [GTP] and has a value greater than 300 s at low [GTP] and a limiting value of 50 s at high [GTP]. We attribute the greater than 300 s lifetime to the complex R*G (where G is ROS G protein) and the 50-s lifetime to free R*. (c) The rate kL increases hyperbolically with [GTP] with a half-maximal value of 56 microM and kL.max = 22 45 s-1. (d) Peak velocity is given by the expression vpeak alpha kL tau P which is consistent with the dependence of kL on [GTP] and the experimental finding that vpeak varies hyperbolically with [GTP]. The minimal model has also allowed us to (a) develop clear definitions of amplification for the light-triggered enzymatic cascade and (b) clarify experimental methods for measuring gain.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2555332 TI - Transient response of rod outer segment cGMP phosphodiesterase to actinic light pulses. II. Detailed quantitative kinetic model. AB - In the accompanying article (Schmidt, J.A., and Yguerabide, J. (1989) J. Biol. Chem. 264, 19790-19803), we presented a minimal quantitative kinetic model with one rate-limiting step for the transient response of rod outer segment (ROS) phosphodiesterase (PDE) to stimulating light pulses of low fractional bleach (linear response range) and showed that the model was in excellent quantitative agreement with experimental results. The model characterizes the PDE response in terms of the specific rate constant of the rate-limiting step, kL, the lifetime of photoactivated rhodopsin, tau R, and the lifetime of activated PDE, tau P, but makes no predictions on how these kinetic parameters should depend on the concentrations of the various reactive species involved in the PDE response to light and does not reveal the nature of the rate-limiting step. However, we established by curve fitting experimental data to theoretical expressions from the model that kL increases hyperbolically with [GTP], tau R decreases with [GTP], and tau P is independent of GTP. In this report we present three detailed kinetic models which make specific quantitative predictions on how the kinetic parameters of the minimal model should depend on nucleotide and G protein concentrations and test the models against experimental data. Each model consists of one rate-limiting step. The first detailed model postulates that the rate limiting step is the dissociation of R*GT into R* and GT (T stands for GTP). The second model postulates that the rate-limiting step is the binding of GTP to R*G, and the third model postulates that the rate-limiting step is the encounter rate of R* and G on the ROS disc membrane. We find that only the first detailed model is consistent with the experimental results as characterized by the minimal model. Using this detailed model we (a) define kL and tau R in terms of more fundamental equilibrium and rate parameters, (b) develop a theory for the systematic evaluation of amplification or gain of the PDE light response from light-stimulated GTP-binding data as well as v(t) versus t graphs, and (c) clarify methods which have been used in the past to evaluate gain experimentally. PMID- 2555333 TI - The oxidation of N-substituted aromatic amines by horseradish peroxidase. AB - The mechanism of N-dealkylation by peroxidases of the Ca2+ indicator quin2 and analogs was investigated and compared with the mechanism of N-dealkylation of some N-methyl-substituted aromatic amines. Nitrogen-centered cation radicals were detected by ESR spectroscopy for all the compounds studied. Further oxidation of the nitrogen-centered cation radicals, however, was dependent upon the structure of the radical formed. In the case of quin2 and analogs, a carbon-centered radical could be detected using the spin trap 5,5-dimethyl-1-pyrroline N-oxide. By using the spin trap 2-methyl-2-nitrosopropane (tert-nitrosobutane), it was determined that the carbon-centered radical was formed due to loss of a carboxylic acid group. This indicated that bond breakage most likely occurred through a rearrangement reaction. Furthermore, extensive oxygen consumption was detected, which was in agreement with the formation of carbon-centered radicals, as they avidly react with molecular oxygen. Thus, reaction of the carbon-centered radical with oxygen most likely led to the formation of a peroxyl radical. The peroxyl radical decomposed into superoxide that was spin trapped by 5,5-dimethyl 1-pyrroline N-oxide and an unstable iminium cation. The iminium cation would subsequently hydrolyze to the monomethyl amine and formaldehyde. In the case of N methyl-substituted aromatic amines, carbon-centered radicals were not detected during the peroxidase-catalyzed oxidation of these compounds. Thus, rearrangement of the nitrogen-centered radical did not occur. Furthermore, little or no oxygen consumption was detected, whereas formaldehyde was formed in all cases. These results indicated that the N-methyl-substituted amines were oxidized by a mechanism different from the mechanism found for quin2 and analogs. PMID- 2555334 TI - Mechanisms of leukotriene E4 partial agonist activity at leukotriene D4 receptors in differentiated U-937 cells. AB - Leukotriene E4 (LTE4) is shown to be a partial agonist of leukotriene D4 (LTD4) in differentiated U-937 cells. The data that support this conclusion are: 1) LTE4 completely displaced [3H]LTD4 from its receptors in U-937 cell membranes. 2) LTE4 induced only 30 +/- 4% of the maximal Ca2+ transient induced by LTD4 in the presence of 1 mM extracellular Ca2+ and 60 +/- 4% of the maximal LTD4 response in the absence of extracellular Ca2+. 3) LTE4 induced only a fraction of the inositol phosphates metabolized by LTD4. Moreover, LTE4 resulted in essentially no production of the inositol 1,4,5-trisphosphate isomer, while LTD4 induced a rapid and substantial transient increase in this isomer. The generation of inositol phosphates by both agonists was unaffected by extracellular Ca2+. 4) The EC50 values for Ca2+ mobilization for LTD4 and LTE4 corresponded with their affinity (Kd values) for the LTD4 receptor. 5) A series of structurally diverse LTD4 receptor antagonists blocked the Ca2+ mobilization responses to LTD4 and LTE4 with identical rank orders of potency. 6) LTE4 acted as an antagonist of LTD4 of potency. 6) LTE4 acted as an antagonist of LTD4 effects when they were coadministered. 7) LTE4 and LTD4 acutely desensitized Ca2+ mobilization to each other. All of the effects of LTE4 are explained by its partial agonist activity at the LTD4 receptor as shown by the following data. 1) Neither LTD4 nor LTE4 had any effect on the agonist activity of fMet-Leu-Phe, LTB4, or platelet-activating factor. 2) None of the above agonists or antagonists to the above receptors affected any of the activities of LTD4 or LTE4. 3) Neither LTD4 nor LTE4 induced desensitization of Ca2+ mobilization to any of the non-LTD4 receptor agonists tested. 4) Under the conditions studied, we have not observed any evidence of multiple subclasses of LTD4 receptors in U-937 cells. LTE4 is a partial agonist of the LTD4 receptor, because it can only couple the LTD4 receptor to a portion of the signaling system available to the receptor when occupied by LTD4. Specifically, LTD4 caused the activation of receptor-operated calcium channels, mobilization of intracellular Ca2+, the activation of phosphatidylinositol phospholipase C, and the liberation of an additional, as yet undefined, intracellular mediator. To do this, LTD4 receptors couple to at least two and perhaps more guanine nucleotide binding proteins. LTE4 is unable to activate the phosphatidylinositol-phospholipase C but can mimic the other effects of LTD4.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2555335 TI - The regulation of the phosphorylation of inositol 1,3,4-trisphosphate in cell free preparations and its relevance to the formation of inositol 1,3,4,6 tetrakisphosphate in agonist-stimulated rat parotid acinar cells. AB - High performance liquid chromatography analysis of supernatants from acid quenched [3H]inositol-labeled parotid acinar cells revealed an inositol pentakisphosphate and three inositol tetrakisphosphates. Two of the latter were identified as the 1,3,4,5 and 1,3,4,6 isomers, whereas the third was probably a mixture of unknown proportions of the 3,4,5,6/1,4,5,6 enantiomeric pair. Methacholine (100 microM) produced a 40-50-fold increase in the levels of inositol trisphosphate (mainly the 1,3,4 isomer) and inositol 1,3,4,5 tetrakisphosphate, but inositol 1,3,4,6-tetrakisphosphate only increased 5-fold. Levels of inositol 3,4,5,6/1,4,5,6-tetrakisphosphate and inositol pentakisphosphate were unaffected by agonist stimulation. Thus, in parotid cells, an agonist-induced increase in both inositol trisphosphate and inositol 1,3,4,6 tetrakisphosphate formation does not result in an increase in the rate of formation of inositol pentakisphosphate. Following the addition of 100 microM atropine to methacholine-stimulated parotid cells, the levels of [3H]inositol 1,3,4,5-tetrakisphosphate fell rapidly, returning to basal levels within 5 min. Inositol trisphosphate was metabolized more slowly and was still elevated 20-fold above basal 5 min after the addition of atropine. Inositol 1,3,4,6 tetrakisphosphate was metabolized much more slowly (t1/2 approximately 15 min). Inositol 1,3,4-trisphosphate metabolism was examined in parotid homogenates as well as in 100,000 x g cytosolic and particulate fractions. Inositol 1,3,4 trisphosphate was both dephosphorylated and phosphorylated. Two inositol tetrakisphosphate products were formed, namely the 1,3,4,6 and 1,3,4,5 isomers. Over 90% of both kinase and phosphatase activities were found in the cytosolic fractions. The ratio of activities of kinase to phosphatase decreased as the levels of inositol 1,3,4-trisphosphate substrate were increased from 1 nM to 10 microM. These data led to the conclusion that the kinetic parameters of the inositol 1,3,4-trisphosphate kinases and phosphatases are such that in stimulated cells, dephosphorylation of inositol 1,3,4-trisphosphate is greatly favored. Inositol 1,3,4-trisphosphate kinase activity was potently inhibited by inositol 3,4,5,6-tetrakisphosphate (IC50 = 0.1-0.2 microM), which leads us to propose that inositol 3,4,5,6-tetrakisphosphate is an endogenous inhibitor of the kinase. PMID- 2555336 TI - The discovery of a 3-phosphomonoesterase that hydrolyzes phosphatidylinositol 3 phosphate in NIH 3T3 cells. AB - Phosphatidylinositol 3-phosphate (PtdIns(3)P), a recently described phospholipid, has been linked to polyoma virus-induced cellular transformation and platelet derived growth factor-mediated mitogenesis. PtdIns(3)P, in contrast to phosphatidylinositol, phosphatidylinositol 4-phosphate (PtdIns(4)P), and phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2), is resistant to hydrolysis by bovine brain phospholipase C gamma. We present here the identification of a phosphomonoesterase activity from the soluble fraction of NIH 3T3 cells which removes the phosphate from the D-3 position of PtdIns(3)P. This enzyme is specific as it has little or no activity on the monoester phosphates of PtdIns(4)P, PtdIns(4,5)P2, or inositol 1,3-bisphosphate and is tentatively designated phosphatidylinositol 3-phosphatase (PtdIns 3-phosphatase). The enzyme does not require added metal ions for activity and is maximally active in the presence of EDTA. It is inhibited by Ca2+, Mg2+, Zn2+, and the phosphatase inhibitor VO4(3-). In addition, there is no phospholipase C activity toward PtdIns(3)P in the soluble fraction of NIH 3T3 cells. In view of the absence of a phospholipase C activity that hydrolyzes PtdIns(3)P, we propose that PtdIns(3)P is not a precursor for a soluble inositol phosphate messenger but that it instead may act directly to control certain cellular processes or as a precursor for other phosphatidylinositols. PtdIns 3-phosphatase may thus terminate a metabolic signal or regulate precursor levels for other phosphatidylinositols that are phosphorylated in the D-3 position. PMID- 2555337 TI - Spectral studies of bovine dopamine beta-hydroxylase. Absence of covalently bound pyrroloquinoline quinone. AB - Bovine dopamine beta-hydroxylase was examined spectroscopically for the presence of covalently bound pyrroloquinoline quinone (PQQ). Pure dopamine beta hydroxylase had a featureless UV-visible spectrum above 300 nm. An equimolar solution of dopamine beta-hydroxylase and exogenously added PQQ (1 PQQ/active site) had a strong absorption maximum at 333 nm. Dialysis removed the added PQQ, indicating that dopamine beta-hydroxylase does not bind PQQ irreversibly. Reaction of dopamine beta-hydroxylase with 6 mM phenylhydrazine in the presence of 15 mM ascorbate caused 96% inactivation within 20 min and did not produce any spectrally detectable amounts of the phenylhydrazone adduct of PQQ, as reported by van der Meer et al. (van der Meer, R.A., Jongejan, J.A., and Duine, J.A. (1988) FEBS Lett. 231, 303-307). The peptide profile of phenylhydrazine inactivated dopamine beta-hydroxylase was monitored at 316 nm and did not reveal any peptides that might contain a PQQ-phenylhydrazone adduct. Thus, the absence of any spectrally detectable PQQ-phenylhydrazone adducts under these conditions demonstrates that the mechanism of phenylhydrazine inactivation does not involve covalent modification of PQQ at the active site of dopamine beta-hydroxylase and provides strong evidence that the native enzyme does not contain PQQ. PMID- 2555338 TI - Agonist-induced destabilization of beta-adrenergic receptor mRNA. Attenuation of glucocorticoid-induced up-regulation of beta-adrenergic receptors. AB - beta-Adrenergic receptor expression and receptor mRNA levels are down-regulated by beta-adrenergic agonists and up-regulated by glucocorticoids. The interaction between these two opposing regulatory pathways was investigated at the levels of receptor and receptor mRNA in DDT1 MF-2 hamster vas deferens cells. Dexamethasone blunted a marked decrease in receptor expression induced by isoproterenol alone, as made visible by indirect immunofluorescence using antireceptor antibodies. Receptor mRNA levels were quantified by DNA-excess solution hybridization. Dexamethasone stimulated a sharp increase in receptor mRNA at 4 h following the addition of steroid in either the absence or the presence of isoproterenol. By 12 h, dexamethasone treatment resulted in a new steady-state level of receptor mRNA double that observed in untreated cells. Isoproterenol blunted the dexamethasone effect observed at 12 h. Cells treated with isoproterenol and dexamethasone in combination displayed a new steady-state level only 30% greater than untreated cells. Measured by nuclear run-on assays, transcription rates of the receptor gene were unaffected in cells challenged with isoproterenol alone. Dexamethasone, in contrast, stimulated a 4-fold increase in beta 2-adrenergic receptor gene transcription. Isoproterenol and dexamethasone in combination promoted a transcription rate comparable to dexamethasone alone. The half-life of receptor mRNA in untreated and dexamethasone-treated cells was 12 h. In contrast, beta adrenergic receptor mRNA half-life declined to 5 h in cells that were treated with isoproterenol in the presence or absence of dexamethasone. Agonist-promoted destabilization and steroid-induced transcription provide mechanisms for the interplay of two opposing pathways controlling receptor mRNA levels. PMID- 2555339 TI - Platelet receptor occupancy with factor IXa promotes factor X activation. AB - To investigate the activated platelet surface as a locus for factor X activation, the functional consequences of factor IXa binding to platelets were studied. The concentration of factor IXa required for half-maximal rates of factor X activation in the presence of factor VIIIa and thrombin-activated platelets was 0.53 nM, which is close to the Kd (0.56 nM) for factor IXa binding to platelets under identical conditions, determined from equilibrium binding studies. In direct comparative experiments, there was a close correspondence between equilibrium binding of factor IXa to thrombin-activated platelets in the presence of factor VIIIa and kinetic determinations of factor X activation rates. Analysis by polyacrylamide gel electrophoresis revealed that 125I-labeled factor IXa bound to platelets was structurally intact and did not form covalent complexes with platelet proteins. Factor IXa active site-inhibited by 5-dimethylaminonaphthalene 1-sulfonyl glutamyl-glycylarginyl chloromethyl ketone was shown to be a competitive inhibitor of factor IXa binding in the absence (Ki = 2.3 nM) and presence (Ki = 0.43 nM) of factor VIIIa and factor X and of factor X activation (Ki = 0.4 nM) by factor IXa in the presence of factor VIIIa, indicating that the generation of factor Xa is not required for factor IXa binding and that factor IXa bound to activated platelets in the presence of factor VIIIa is closely coupled with rates of factor X activation. We conclude that factor IXa bound tightly to a platelet receptor in the presence of factor VIIIa is the enzyme active in factor X activation. PMID- 2555340 TI - Purification and properties of vacuolar membrane proton-translocating inorganic pyrophosphatase from mung bean. AB - Inorganic pyrophosphatase was purified from the vacuolar membrane of mung bean hypocotyl tissue by solubilization with lysophosphatidylcholine and QAE-Toyopearl chromatography. The molecular mass on sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 73,000 daltons. Among the amino-terminal first 30 amino acids are 25 nonpolar hydrophobic residues. For maximum activity, the purified pyrophosphatase required 1 mM Mg2+ and 50 mM K+. The enzyme reaction was stimulated by exogenous phospholipid in the presence of detergent. Excess pyrophosphate as well as excess magnesium inhibited the pyrophosphatase. The enzyme reaction was strongly inhibited by ATP, GTP, and CTP at 2 mM, and the inhibition was reversed by increasing the Mg2+ concentration. An antibody preparation raised in a rabbit against the purified enzyme inhibited both the reactions of pyrophosphate hydrolysis of the purified preparation and the pyrophosphate-dependent H+ translocation in the tonoplast vesicles. N,N' Dicyclohexylcarbodiimide became bound to the purified pyrophosphatase and inhibited the reaction of pyrophosphate hydrolysis. It is concluded that the 73 kDa protein in vacuolar membrane functions as an H+-translocating inorganic pyrophosphatase. PMID- 2555341 TI - Molecular cloning of a novel mammalian calcium-dependent protease distinct from both m- and mu-types. Specific expression of the mRNA in skeletal muscle. AB - Two types of calcium-dependent protease with distinct calcium requirements (termed muCANP and mCANP) are known in mammalian tissues. These two isozymes consist of different large (80-kDa) subunits (mu- or m-types) and identical small (30-kDa) subunits. By screening human and rat muscle cDNA libraries with a cDNA probe for the chicken CANP large subunit, which has a structure similar to both the mammalian mu- and m-types, a cDNA clone encoding a novel member of the CANP large subunit family was obtained. The encoded protein (designated "p94") consists of 821 amino acid residues (Mr 94,084) and shows significant sequence homology with both human mu-type (54%) and m-type (51%) large subunits. p94 can be divided into four domains (I-IV) as reported for the CANP large subunit family. Domains II and IV are potential cysteine protease and calcium-binding domains, respectively, and have sequences homologous to the corresponding domains of other CANP large subunits. However, domain I of p94 is significantly different from others. Moreover, p94 contains two unique sequences of 62 and 77 residues in domains II and III, respectively. In contrast to the ubiquitous expression of mu- and m-types, Northern blot analysis revealed that the mRNA for p94 exists only in skeletal muscle with none detected in other tissues including heart muscle and smooth muscles such as intestine. PMID- 2555342 TI - Chemotactic peptide induces cAMP elevation in human neutrophils by amplification of the adenylate cyclase response to endogenously produced adenosine. AB - The transient increase in human neutrophil cAMP levels induced by the chemoattractant N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP) is shown to be caused by amplification of adenylate cyclase response to endogenously produced adenosine. The FMLP-stimulated increase in neutrophil cAMP was potentiated markedly by a nonmethylxanthine cAMP phosphodiesterase inhibitor (Ro 20-1724). By inhibiting the degradation of newly formed cAMP, Ro 20-1724 rendered the FMLP induced cAMP elevation persistent rather than transient. The role of endogenously produced adenosine in this phenomenon is demonstrated by the ability of either adenosine deaminase or theophylline, an adenosine receptor antagonist, to prevent FMLP-stimulated cAMP elevation. The general nature of the FMLP-potentiated cAMP response is indicated by the finding that FMLP-treated neutrophils, in the presence of exogenously supplied adenosine deaminase, exhibited augmented cAMP generation in response to three different types of receptor agonists: 2 chloroadenosine, prostaglandin E1, and L-isoproterenol. Moreover, like the neutrophil cAMP increase caused by FMLP alone, the ability of FMLP to augment cAMP response to 2-chloroadenosine in adenosine deaminase-treated cells was short lived and declined after 1.0 min of exposure to FMLP. Preincubation of neutrophil suspensions with the adenylate cyclase inhibitor SQ 22,536 completely prevented FMLP-induced cAMP generation. Furthermore, when neutrophil suspensions were preincubated with concentrations of Ro 20-1724, which apparently maximally inhibit cAMP phosphodiesterase, a 30-s incubation with FMLP still resulted in substantially elevated cAMP levels. It therefore appears that FMLP raises cAMP by activating adenylate cyclase rather than inhibiting cAMP phosphodiesterase. PMID- 2555343 TI - Phosphatidylinositol 3-kinase and its novel product, phosphatidylinositol 3 phosphate, are present in Saccharomyces cerevisiae. AB - The metabolism of polyphosphoinositides has been shown to be an important factor in controlling the proliferation of Saccharomyces cerevisiae. The monophosphate form of phosphatidylinositol has been assumed to be phosphatidylinositol 4 phosphate (PI-4-P). Recent evidence from our laboratory has established that a phosphatidylinositol (PI) kinase, which phosphorylates the D-3 position of the inositol ring (PI 3-kinase), is associated with many activated protein-tyrosine kinases and may play an important role in the signaling of cell proliferation (Auger, K. R., Serunian, L. A., Soltoff, S. P., Libby, P., and Cantley, L. C. (1989) Cell 57, 167-175). To determine the evolutionary conservation of this enzymatic activity, we investigated its presence in yeast. In vitro PI kinase assays of yeast cell homogenates demonstrated that PI 3-kinase activity was present. Preliminary biochemical characterization of the activity suggested that it was quite different from the mammalian enzyme yet catalyzed the same reaction, i.e. phosphorylating the D-3 hydroxyl position of the inositol ring of phosphatidyl-myo-inositol. [3H]Inositol labeling of intact yeast cells with the subsequent extraction, deacylation, and high performance liquid chromatography analysis of the lipids demonstrated that PI-3-P was as abundant as the PI-4-P isomer. The conservation of the enzymatic activity from yeast to man suggests that it has an important functional role in the cell cycle. PMID- 2555344 TI - Metabolism of inositol pentakisphosphate to inositol hexakisphosphate in Xenopus laevis oocytes. AB - The formation and metabolism of inositol pentakis-and hexakisphosphates (InsP5 and InsP6) were investigated in Xenopus laevis oocytes. After [3H]inositol injection, [3H]InsP5 and subsequently [3H]Insp6 increased progressively over 72 h. In intact oocytes, [3H]InsP5 was progressively converted to [3H]InsP6 from 6 to 72 h of incubation and was not metabolized to lower inositol phosphates. In contrast, [3H]InsP6 remained unmetabolized for up to 72 h. These data are consistent with the kinetics of the increases in [3H]InsP5 and [3H]InsP6 in [3H]inositol-labeled oocytes. The highly phosphorylated inositols showed significant changes during oogenesis and maturation. In oocytes incubated for 48 h after [3H]inositol injection, the radioactive incorporation into polyphosphoinositols increased progressively from stage 3 to stage 6, with 5- and 6-fold rises (cpm/mg protein) for [3H]InsP5 and [3H]InsP6, respectively. These developmental changes were associated with 5-fold increases in [3H]inositol tetrakisphosphate between stages 3 and 6 of oogenesis. Induction of oocyte maturation by progesterone (1 microM) during the last 12 of a 36-h incubation with [3H]inositol doubled the levels of [3H]InsP6 relative to [3H]InsP5, suggesting that the activity of inositol pentakisphosphate kinase increases during maturation. These results provide direct evidence for metabolic conversion of InsP5 to InsP6 in animal cells and show that the higher inositol polyphosphates, unlike the lower phosphoinositols, are extraordinarily stable. These species increase markedly during ovum development and may play a regulatory role in oogenesis and maturation. PMID- 2555345 TI - Cell line-dependent response of the enhancer element of simian virus 40 to transactivator p40tax encoded by human T-cell leukemia virus type I. AB - The trans-acting transcriptional factor p40tax of human T-cell leukemia virus type I (HTLV-I) can activate expression not only of its own viral genes but also of other viral and cellular genes. We examined the cis-acting sequences in the simian virus 40 (SV40) enhancer required for its activation by HTLV-I p40tax. Experiments with chloramphenicol acetyltransferase constructs bearing the SV40 enhancer elements revealed that p40tax-dependent transactivation of the SV40 enhancer is mediated through the C element that contains the typical sequence for binding of a nuclear factor NF-kappa B. Activation of the C element by p40tax was seen in a limited set of cell lines as was also seen with the whole enhancer of SV40. Binding of NF-kappa B or NF-kappa B-like factor to the SV40 C element increased when p40tax was expressed. The fact that HTLV-I p40tax utilizes a cellular regulatory mechanism to transactivate the SV40 enhancer in a cell dependent manner may have implications for the pathogenesis of adult T-cell leukemia. PMID- 2555347 TI - The mitochondrial targeting function of randomly generated peptide sequences correlates with predicted helical amphiphilicity. AB - A pool of oligonucleotides encoding a start methionine and nine random amino acids was inserted at the 5'-end of the gene for the yeast cytochrome oxidase subunit IV lacking its own mitochondrial targeting sequence. Approximately one quarter of the randomly generated sequences targeted subunit IV to its correct intramitochondrial location in vivo. Sequence analysis of 89 randomly generated sequences showed that their efficiencies as mitochondrial targeting signals correlated with the potential to fold into an amphiphilic alpha-helix. Functional targeting sequences were enriched in arginine and isoleucine residues but contained few aspartate, glutamate, and proline residues. Nonfunctional sequences predicted to have significant helical amphiphilicity often had at least one acidic or multiple helix-breaking residues that would be expected to interfere with targeting functioning. These results support the hypothesis that the signal for targeting a protein into the mitochondrial matrix is usually a positively charged amphiphilic helix. PMID- 2555346 TI - Purification and partial characterization of membrane-homing receptors in two cloned murine hemopoietic progenitor cell lines. AB - Selective seeding of bone marrow by intravenously transplanted hemopoietic cells depends on the homing receptors of these cells. The receptors are membrane lectins with galactosyl and mannosyl specificities. To purify these lectins, cell membrane was fractionated from two cloned multipotential (B6STU) and bipotential (FDCP-1) hemopoietic progenitor cells. The membrane was solubilized and its proteins were labeled with 125I. The proteins were subjected to affinity column chromatography using galactosyl and mannosyl groups linked to agarose beads. Elution with D-galactose or D-mannose led to specific elution of a single sharp radioactive peak which constituted a constant fraction of membrane proteins. This peak was studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis or by disuccinimidyl suberate cross-linking technique and appeared to have a Mr of 110,000. Under reducing conditions, it consisted of two components with a Mr of 87,000 and 23,000. Treatment with endoglycosidase F indicated about 5% carbohydrate content. Purification of these homing receptors has opened an avenue for the development of immunologic and molecular biologic probes that may help further elucidate the mechanism of homing regulation. PMID- 2555348 TI - Hydroxyl radical production during oxidative deposition of iron in ferritin. AB - The chemistry of oxidative deposition of iron(III) in ferritin and apoferritin is poorly understood. This study was undertaken to look for radicals formed as the hydrous ferric oxide core is developed from Fe(II) and O2. Radicals were observed indirectly by using the spin-trapping reagent N-tert-butyl-alpha-phenylnitrone (PBN) at room temperature and directly by measuring ESR spectra of frozen solutions at 77 K. In both instances, radical production was inhibited by the hydroxyl radical scavenging agents dimethyl sulfoxide, thiourea, and mannitol and enhanced by the addition of hydrogen peroxide. These findings strongly suggest that hydroxyl radical, produced from the iron-catalyzed Haber-Weiss reaction, is a by-product of core formation in ferritin and is a precursor to the observed radicals. The yield of ESR-observable and spin-trapped radicals is quite low, being at the micromolar level when millimolar concentrations of ferrous ion are employed. Furthermore, radical production appears to be confined to the interior of the ferritin molecule, where cellular components would be protected from the oxygen-derived toxic effects of iron. It is postulated that hydroxyl radical medicated oxidative damage to the protein, a process that may contribute to the formation of hemosiderin from ferritin, leads to the observed radicals. By serving as a sink for hydroxyl radical, the protein shell may therefore efficiently minimize damage to other biomolecules in the cell. PMID- 2555349 TI - Mutant vitamin D receptors which confer hereditary resistance to 1,25 dihydroxyvitamin D3 in humans are transcriptionally inactive in vitro. AB - Hereditary 1,25-dihydroxyvitamin D3-resistant rickets is a human syndrome that arises as a result of heterogeneous molecular defects in the vitamin D3 receptor. Recent studies have identified single unique point mutations within the second or third exons that encode the DNA-binding domain of the vitamin D receptor (VDR) gene in two families with this syndrome. In the experiments reported here, these mutations were introduced into the normal VDR cDNA by site-directed mutagenesis and the mutant products evaluated for hormone, nuclear, and DNA-binding characteristics. Each mutant VDR was expressed in COS-1 cells at equivalent levels, and saturation analysis of cell cytosol revealed normal affinity for the 1,25-dihydroxyvitamin D3 hormone. Incubation of transfected cells with radiolabeled hormone followed by lysis and extraction suggests a lowered salt dependence for solubilization of the mutant VDR. Concomitantly, mutant receptors exhibited reduced affinity for immobilized calf thymus DNA. While cotransfection of the wild type receptor together with a vitamin D-inducible (osteocalcin) chloramphenicol acetyltransferase reporter gene construction in CV-1 cells resulted in strong induction by 1,25-dihydroxyvitamin D3, neither mutant receptor was capable of directing significant activity either as a function of receptor or hormone concentration. These data suggest that the unique point mutations identified in each of these two families are responsible not only for the phenotype originally ascribed to the abnormal receptor but also severely compromise each protein's ability to activate transcription. PMID- 2555351 TI - Deletion of ant in Escherichia coli reveals its function in adaptation to high salinity and an alternative Na+/H+ antiporter system(s). AB - We have deleted the chromosomal ant gene from Escherichia coli by substitution with the kan gene, which encodes kanamycin resistance. The delta ant strains obtained cannot adapt to high sodium concentrations (700 mM, pH 6.8), which do not affect the wild type. The Na+ sensitivity of delta ant is pH dependent, increasing at alkaline pH. Thus at pH 8.5, 100 mM NaCl retard growth of delta ant with no effect on the wild type. The delta ant strains also cannot challenge the toxic effects of Li+ ions, a substrate of the Na+/H+ antiporter system. However, growth of these strains is normal on carbon sources which require Na+ ions for transport and growth. Moreover, antiporter activity, as measured in everted membrane vesicles, is not significantly impaired. A detailed analysis of the remaining antiporter activity in a delta ant strain reveals kinetic properties which differ from those displayed by the ant protein: (a) Km for transport of Li+ ions is about 15 times higher and (b) the activity is practically independent of intracellular pH. Our results demonstrate the presence of an alternative Na+/H+ antiporter(s) in E. coli, additional to ant system. PMID- 2555350 TI - Structural determinants of the factor IX molecule mediating interaction with the endothelial cell binding site are distinct from those involved in phospholipid binding. AB - Previous studies have indicated that Factor IX/IXa interacts in a specific and high affinity manner with a binding site on the endothelial cell surface. In this study, the contributions of the gamma-carboxyglutamic acid-containing (GLA) and growth factor domains to the finding of Factor IX to the endothelium were assessed. While GLA-containing peptides from Factors IX, X, and prothrombin were inhibitors of 125I-Factor IX-endothelial cell binding, the GLA peptide from Factor IX was about 250-800-fold more effective than those from prothrombin and Factor X, respectively. In contrast to its relative efficacy as an inhibitor of Factor IX-cell surface interaction, the Factor IX-GLA peptide neither bound to lipid vesicles nor inhibited Factor IX-lipid interaction. A synthetic peptide comprising the entire first epidermal growth factor (EGF) exon was also an inhibitor of 125I-Factor IX-endothelial cell binding, although it did not interact with lipid vesicles. Experiments with synthetic peptides comprising each of the three loops of the first EGF domain or the entire first EGF region with specific substitutions indicated the importance of determinants in both the first and probably third loops for Factor IX-endothelial interaction. In contrast, the second loop of the first EGF domain and the first loop of the second EGF exon are probably not involved in Factor IX-endothelial interaction based on their inability to block 125I-Factor IX binding to cells. These results indicate that determinants in both the GLA and the first EGF domain contribute to the specific binding of Factor IX to the endothelial cell surface and that structural requirements for Factor IX-cell surface interaction are distinct from those for Factor IX binding to lipids. PMID- 2555352 TI - Synthetic inositol trisphosphate analogs and their effects on phosphatase, kinase, and the release of Ca2+. AB - A series of inositol 1,4,5-trisphosphate (IP3) analogs and positional isomers was examined to explore the structure-activity relationships among IP3 5-phosphatase, IP3 3-kinase, and the release of Ca2+. All analogs with additional groups on the 2nd position of IP3 inhibited the hydrolysis of [5-32P]IP3 catalyzed by erythrocyte ghosts, with a lower Ki value than seen with IP3. IP3 dehydroxylated at the 2nd position also had a lower Ki, while 2,4,5-IP3 or cyclic(1:2), 4,5-IP3 had higher Ki values. Among these compounds 2-deoxy-IP3 was as potent as IP3 in inhibiting the phosphorylation by [3H] IP3-3-kinase in rat brain cytosol. The other compounds, except for 2,4,5-IP3 inhibited the phosphorylation, however, 2 30 times higher concentrations were required. By lowering free Ca2+, the concentrations required for half-maximal inhibition were low, while those of IP3, 2-deoxy-IP3, and positional isomers remained unchanged. These compounds acted as full agonists in releasing Ca2+ from permeabilized macrophages, although 1.6-50 fold higher concentrations than IP3 were required. These compounds also inhibited the binding of [3H]IP3 to rat cerebellum and bovine adrenal cortex microsomes, but the potencies were 2.9-33 times less than that of IP3. Thus, the 2nd position of IP3 can be modified with only a slight loss of biological activity. PMID- 2555353 TI - A synthetic linear decapeptide binds to the atrial natriuretic peptide receptors and demonstrates cyclase activation and vasorelaxant activity. AB - A linear decapeptide, [cyclohexylalanine 106]ANP-(105-114)NH2 (1), where ANP is atrial natriuretic peptide, was prepared by solid phase synthesis and purified by reverse-phase liquid chromatography. This novel peptide was found to bind to ANP receptors in rabbit lung membranes, to stimulate cGMP production in various tissues, and to fully relax precontracted rabbit aorta in a dose-dependent fashion. The potency of 1 in the various in vitro assays varies between one twentieth and one-eightieth of the potency of the reference peptide, the 24-mer rat ANP-(103-126). The linear decapeptide 1, which encompasses amino acid residues from the rat ANP sequence (105-114), features a cyclohexylalanine residue instead of the phenylalanine 106 residue in the hormone sequence, a free sulfhydryl function at the N-terminal cysteine 105, and a carboxamide C terminus. Its disulfide dimer 6 was active in the rabbit aorta assay while the S-methyl cysteine 7 analogue was not active in the same assay at similar concentrations. The decapeptide 1 is of particular significance because it is the shortest analogue reported to date endowed with agonistic activity at the guanylate cyclase-coupled ANP receptor. In particular, it is interesting to compare its structure to the structures of other short linear analogues of ANP which are totally devoid of the ability to stimulate particulate guanylate cyclase activity. PMID- 2555354 TI - [3H]bumetanide binding in vascular endothelial cells. Quantitation of Na-K-Cl cotransporters. AB - Vascular endothelial cells previously have been shown to possess a prominent Na-K Cl cotransport system which mediates a K+ influx of approximately 20 mumols/g of protein/min. Endothelial cell cotransport has also been shown to be regulated by a variety of vasoactive agents and their second messengers, suggesting that the transport system may have an important role in endothelial cell function. In the present study we investigated the possibility that the high level of cotransport in these cells is due to a large number of Na-K-Cl cotransporters in the plasma membrane. This was done by evaluating specific saturable binding of [3H]bumetanide to cultured bovine aortic endothelial cells. We found a maximal [3H]bumetanide binding of 0.83 pmol/mg protein with a dissociation constant of 0.13 microM. From these data, the number of [3H]bumetanide binding sites/endothelial cell was determined to be approximately 230,000, and the turnover number for cotransport activity was calculated to be 300 K+ ions/site/s. These findings indicate that endothelial cells do indeed exhibit a large number of Na-K-Cl cotransporters/cell relative to other cell types. We also investigated the effects on [3H]bumetanide binding of agents known to modulate Na-K-Cl cotransport activity. Saturable binding of [3H]bumetanide was found to be reduced significantly by treatment of the cells with 8-bromo-cyclic AMP, 8-bromo-cyclic GMP, phorbol esters, norepinephrine, or rat atriopeptin III, all of which have been shown to inhibit Na-K-Cl cotransport-mediated K+ influx. PMID- 2555355 TI - Purification of a ubiquitin protein peptidase from yeast with efficient in vitro assays. AB - In eukaryotic cells ubiquitin is synthesized as a polyubiquitin protein or as a protein fused at the carboxyl terminus to other polypeptides. An enzyme activity, ubiquitin protein peptidase, has been proposed to process these precursors by cleaving the peptide bond between adjoining ubiquitin molecules or between ubiquitin and the fused peptides. Using the cleavage of a 35S-labeled yeast ubiquitin protein fused to a synthetic 38-residue peptide obtained by in vivo metabolic labeling in Escherichia coli in an expression system based on the interaction of bacteriophage T7 RNA polymerase and its promoter, it is possible to detect a processing activity in soluble yeast extract. The specificity of the cleavage suggests this activity could be the in vivo processing activity for various ubiquitin precursor proteins in yeast cells. A similarly labeled ubiquitin protein fused to one cysteine residue was also utilized to detect an activity capable of removing a single cysteine residue from ubiquitin in a soluble extract. Employing assays based on the cleavage of labeled ubiquitin protein fusions, a ubiquitin protein peptidase activity from Saccharomyces cerevisiae was purified about 15,000-fold to yield a protein mixture consisting of only a few protein species. The major protein band which comigrated with the activities in in vitro assays has an apparent molecular weight of 29,000 when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Two other protein species, about 20,000 and 10,000 in molecular weight, also comigrated with the in vitro activities throughout the purification procedure. Though our most purified protein fraction was shown to cleave various artificial ubiquitin protein fusions under our experimental conditions, it cannot cleave a ubiquitin dimer protein, suggesting the existence of functionally distinct ubiquitin protein peptidases. Our experimental protocol for preparing various labeled ubiquitin protein precursors provides a means to explore various processing enzymes existing in cells. The same protocol may also be adapted to prepare substrates for the study of other specific protein processing enzymes. PMID- 2555356 TI - A transfected m1 muscarinic acetylcholine receptor stimulates adenylate cyclase via phosphatidylinositol hydrolysis. AB - The m1 muscarinic acetylcholine receptor gene was transfected into and stably expressed in A9 L cells. The muscarinic receptor agonist, carbachol, stimulated inositol phosphate generation, arachidonic acid release, and cAMP accumulation in these cells. Carbachol stimulated arachidonic acid and inositol phosphate release with similar potencies, while cAMP generation required a higher concentration. Studies were performed to determine if the carbachol-stimulated cAMP accumulation was due to direct coupling of the m1 muscarinic receptor to adenylate cyclase via a GTP binding protein or mediated by other second messengers. Carbachol failed to stimulate adenylate cyclase activity in A9 L cell membranes, whereas prostaglandin E2 did, suggesting indirect stimulation. The phorbol ester, phorbol 12-myristate 13-acetate (PMA), stimulated arachidonic acid release yet inhibited cAMP accumulation in response to carbachol. PMA also inhibited inositol phosphate release in response to carbachol, suggesting that activation of phospholipase C might be involved in cAMP accumulation. PMA did not inhibit prostaglandin E2-, cholera toxin-, or forskolin-stimulated cAMP accumulation. The phospholipase A2 inhibitor eicosatetraenoic acid and the cyclooxygenase inhibitors indomethacin and naproxen had no effect on carbachol-stimulated cAMP accumulation. Carbachol stimulated cAMP accumulation was inhibited with TMB-8, an inhibitor of intracellular calcium release, and W7, a calmodulin antagonist. These observations suggest that carbachol-stimulated cAMP accumulation does not occur through direct m1 muscarinic receptor coupling or through the release of arachidonic acid and its metabolites, but is mediated through the activation of phospholipase C. The generation of cytosolic calcium via inositol 1,4,5 trisphosphate and subsequent activation of calmodulin by m1 muscarinic receptor stimulation of phospholipase C appears to generate the accumulation of cAMP. PMID- 2555357 TI - A functional cell surface type receptor is required for the early action of 1,25 dihydroxyvitamin D3 on the phosphoinositide metabolism in rat enterocytes. AB - 1,25-Dihydroxyvitamin D3 (1,25-(OH)2-D3) (0.1 pM to 2 nM) induces a concomitant very rapid (within 5 s) and transient release of inositol trisphosphate and diacylglycerol in enterocytes from 3-month-old rats. This high level is not maintained but declines to a lower level 60 s after stimulation. The stimulating effect is dose-dependent and biphasic with a maximum effect at 10 pM. 1,25-(OH)2 D3 induces a rapid (within 10 s) accumulation of inositol bisphosphate and its effect on inositol monophosphate is delayed (120 s). The primary action of 1,25 (OH)2-D3 is to initiate hydrolysis of phosphatidyl 4,5-bisphosphate to yield inositol trisphosphate and diacylglycerol. In contrast, 1,25-(OH)2-D3, for any of the concentrations and the incubation time periods tested, has no effect on inositol trisphosphate and diacylglycerol formation in enterocytes from 16-day old rats at a time when specific binding sites for 1,25-(OH)2-D3 cannot be detected. In conclusion, the early (within 5-60 s) effects of 1,25-(OH)2-D3 on small intestinal epithelium may be mediated via the phosphoinositide transduction system and require the presence of functioning cell membrane-type receptors. PMID- 2555358 TI - Molecular cloning and expression of cDNA encoding a peripheral-type benzodiazepine receptor. AB - This report describes the cloning of a full length cDNA encoding PKBS, a protein of approximately 17 kDa associated with peripheral-type benzodiazepine binding sites. Cyanogen bromide digestion of purified PKBS yielded several peptide fragments which were subjected to gas-phase sequencing. Based on these partial amino acid sequences, oligonucleotide probes were used to screen a rat adrenal cDNA library. Several hybridizing clones were isolated which were found to contain overlapping sequences. The longest cDNA spanned 781 base pairs and specified an open reading frame of 169 amino acids which matched all of the peptide sequences. Northern analysis with this PKBS cDNA probe in different rat tissues revealed one RNA species of approximately 850 nucleotides exhibiting relative abundances qualitatively comparable with the densities of peripheral type benzodiazepine binding sites in each tissue. The PKBS cDNA was cloned into an eukaryotic expression vector placing it under transcriptional control of the beta-globin promoter and SV40 enhancer. Transfection of the transformed human kidney 293 cell line with this recombinant vector resulted in stoichiometric increases of about 900 fmol/mg of protein in binding capacities for Ro5-4864 (4' chlorodiazepam) and PK 11195, two peripheral-type benzodiazepine ligands. These increases were accounted for by the expression of binding sites with approximate dissociation constants of 5 nM for PK 11195 and 8 nM for Ro5-4864, thereby distinguishing the expressed binding sites as being characteristic of the receptor from rat origin rather than of the host human-derived cell line. The rank order of potency of different ligands to compete against [3H]Ro5-4864 binding in the transfected cells was PK 11195 greater than Ro5-4864 greater than diazepam greater than protoporphyrin IX greater than clonazepam, consistent with the specificity characteristic of rat peripheral-type benzodiazepine binding sites. These studies suggest that PKBS comprises binding domains for benzodiazepines and isoquinoline carboxamides and hence is apparently responsible for the manifestation of peripheral-type benzodiazepine recognition sites. PMID- 2555359 TI - Regulation of platelet-activating factor receptors in rat Kupffer cells. AB - Ligand binding studies indicate that 1-O-alkyl-2-acetyl-sn-glycero-3 phosphocholine (AGEPC) down-regulates its own receptors on the plasma membrane of isolated rat Kupffer cells but has no significant effect on the binding affinity of the receptor for AGEPC. Exposure of isolated rat Kupffer cells to 10(-8) and 10(-6) M AGEPC resulted in a rapid, time-dependent reduction in the number of cell surface AGEPC receptors to a new steady state concentration (54.1 +/- 5.0% and 38.6 +/- 5.4% of control, respectively). During the observation period (6 h), the half-time of surface AGEPC receptors was about 60 and 45 min in the presence of 10(-8) and 10(-6) M AGEPC, respectively. Both the rate of loss and the maximal loss of the receptors were dependent upon the AGEPC concentration. With receptor synthesis inhibited by cycloheximide in the absence of AGEPC, the half-time of the surface AGEPC receptor was about 4 h, suggesting that AGEPC receptors are not recycled and that the loss of AGEPC receptors from the plasma membrane is accelerated by AGEPC binding. When incubated with Kupffer cells at 37 degrees C for 3 h, 1-O-alkyl-2-lyso-sn-glycero-3-phosphocholine (1.0 microM), an inactive metabolite of AGEPC, did not cause the loss of AGEPC receptors. Under the same conditions, AGEPC antagonists such as BN52021 (2 x 10(-5) M) or U66985 (2 x 10( 5) M) alone had no effect (97.0 +/- 3.9% of control for BN52021) or only a relatively slight effect (78.4 +/- 1.8% for U66985) on the number of surface AGEPC receptors. However, AGEPC antagonists inhibited the AGEPC-induced down regulation of AGEPC receptors in a concentration-dependent manner, suggesting that the AGEPC-induced down-regulation of AGEPC receptors is a receptor-mediated process. The AGEPC-mediated decrease in receptor number on rat Kupffer cells is reversible. Upon removing AGEPC from the culture medium, about 67% of the lost receptors were replaced within 2 h. Cycloheximide, an inhibitor of protein synthesis, prevented the restoration of the AGEPC receptors. Similar results were obtained when Kupffer cells were incubated with Pronase followed by removing Pronase and reincubating the cells with or without cycloheximide. These observations suggest that the restored AGEPC receptor is newly synthesized rather than recycled. The present study demonstrates that under non-stimulatory (i.e. in the absence of AGEPC) conditions AGEPC receptors are lost from the plasma membrane and are reformed in the cells continuously.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2555360 TI - Spectroscopic characterization of secondary amine mono-oxygenase. Comparison to cytochrome P-450 and myoglobin. AB - Secondary amine mono-oxygenase from Pseudomonas aminovorans catalyzes the NAD(P)H and dioxygen-dependent N-dealkylation of secondary amines to yield a primary amine and an aldehyde. Heme iron, flavin, and non-heme iron prosthetic groups are known to be present in the oligomeric enzyme. The N-dealkylation reaction is also catalyzed by the only other heme-containing mono-oxygenase, cytochrome P-450. In order to identify the heme iron axial ligands of secondary amine mono-oxygenase so as to better define the structural requirements for oxygen activation by heme enzymes, we have investigated the spectroscopic properties of the enzyme. The application of three different spectroscopic techniques, UV-visible absorption, magnetic circular dichroism and electron paramagnetic resonance, to study eight separate enzyme derivatives has provided extensive and convincing evidence for the presence of a proximal histidine ligand. This conclusion is based primarily on comparisons of the spectral properties of the enzyme with those of parallel derivatives of myoglobin (histidine proximal ligand) and P-450 (cysteinate proximal ligand). Spectral studies of ferric secondary amine mono-oxygenase as a function of pH have led to the proposal that the distal ligand is water. Deprotonation of the distal water ligand occurs upon either raising the pH to 9.0 or substrate (dimethylamine) binding. In contrast, the deoxyferrous enzyme appears to have a weakly bound nitrogen donor distal ligand. Initial spectroscopic studies of the iron-sulfur units in the enzyme are interpreted in terms of a pair of Fe2S2 clusters. Secondary amine mono-oxygenase is unique in its ability to function as cytochrome P-450 in activating molecular oxygen but to do so with a myoglobin-like active site. As such, it provides an important system with which to probe structure-function relations in heme-containing oxygenases. PMID- 2555361 TI - Tumor necrosis factor modulates epidermal growth factor receptor phosphorylation and kinase activity in human tumor cells. Correlation with cytotoxicity. AB - Tumor necrosis factor (TNF) is a cytokine which induces cytotoxicity in some but not all tumor cells. Initial studies of five tumor cell lines demonstrated that TNF was able to rapidly (within 30 min) modulate tyrosine protein kinase activity of epidermal growth factor (EGF) receptors on tumor cell lines which were sensitive to the cytotoxic effects of TNF but not alter EGF receptor kinase activity in TNF-resistant tumor cells. Two tumor cell lines (ME-180 cervical carcinoma and T24 bladder carcinoma) which have been shown to express similar TNF binding characteristics but differ in their sensitivity to the cytotoxic actions of TNF were chosen for further characterization. Treatment of TNF-sensitive ME 180 cells with 1 nM TNF resulted in a 3-fold stimulation of EGF receptor tyrosine protein kinase activity within 10 min which correlated with increased phosphorylation of EGF receptor protein itself. In addition, dose-response studies indicate that similar concentrations of TNF modulate both ME-180 cell growth and EGF receptor kinase activity. Treatment of TNF-resistant T24 cells showed that TNF had no significant effect on their growth, EGF receptor tyrosine protein kinase activity, or phosphorylation of EGF receptor protein although EGF receptor kinase activity was stimulated by EGF. Quantitation of receptors expressed on the surface of ME-180 and T24 cells demonstrated a 3-fold difference between the number of EGF-binding sites on T24 (100,000) versus ME-180 cells (300,000), suggesting the relative abundance of EGF receptor does not solely account for differential effects of TNF on EGF receptor activation in these two cell lines. Phosphoamino acid analysis of EGF receptor from 32P-equilibrated ME 180 cells demonstrated that TNF-induced phosphorylation of amino acids which was quantitatively similar to that of EGF but distinct from the effects of phorbol ester. However, unlike EGF, TNF was unable to stimulate EGF receptor kinase activity in ME-180 cell lysates. The kinetics of EGF receptor activation and the metabolic consequence of activation of EGF receptor activity by TNF appear to be distinct from those induced by EGF. These results suggest that TNF-induced modulation of EGF receptor occurs through a unique mechanism and may play a role in the cytotoxic actions of TNF. PMID- 2555362 TI - Purification and characterization of Rad3 ATPase/DNA helicase from Saccharomyces cerevisiae. AB - The Rad3 ATPase/DNA helicase was purified to physical homogeneity from extracts of yeast cells containing overexpressed Rad3 protein. The DNA helicase can unwind duplex regions as short as 11 base pairs in a partially duplex circular DNA substrate and does so by a strictly processive mechanism. On partially duplex linear substrates, the enzyme has a strict 5'----3' polarity with respect to the single strand to which it binds. Nicked circular DNA is not utilized as a substrate, and the enzyme requires single-stranded gaps between 5 and 21 nucleotides long to unwind oligonucleotide fragments from partially duplex linear molecules. The enzyme also requires duplex regions at least 11 base pairs long when these are present at the ends of linear molecules. Rad3 DNA helicase activity is inhibited by the presence of ultraviolet-induced photoproducts in duplex regions of partially duplex circular molecules. PMID- 2555363 TI - An alpha-glucose-1-phosphate phosphodiesterase is present in rat liver cytosol. AB - UDP-glucose:glycoprotein glucose-1-phosphotransferase (Glc-phosphotransferase) catalyzes the transfer of alpha-Glc-1-P from UDP-Glc to mannose residues on acceptor glycoproteins. The predominant acceptor for this transfer in both mammalian cells and Paramecium is a cytoplasmic glycoprotein of 62-63 kDa. When cytoplasmic proteins from rat liver were fractionated by preparative isoelectric focusing following incubation of a liver homogenate with the 35S-labeled phosphorothioate analogue of UDP-Glc ([beta-35S]UDP-Glc), the acceptor was found to have a pI of about 6.0. This fraction, when not labeled prior to the focusing, became very heavily labeled when mixed with [beta-35S]. UDP-Glc and intact liver microsomes, a rich source of the Glc-phosphotransferase. In addition, it was observed that the isoelectric fractions of the cytosol having pI values of 2-3.2 contained a degradative activity, alpha-Glc-1-P phosphodiesterase, that was capable of removing alpha-Glc-1-P, monitored through radioactive labeling both in the sugar and the phosphate, as an intact unit from the 62-kDa acceptor. Identification of the product of this cleavage was substantiated by its partial transformation to UDP-Glc in the presence of UTP and UDP-Glc pyrophosphorylase. The alpha-Glc-1-P phosphodiesterase had a pH optimum of 7.5 and was not effectively inhibited by any of the potential biochemical inhibitors that were tested. Specificity for the Glc-alpha-1-P-6-Man diester was suggested by the diesterase's inability to degrade UDP-Glc or glucosylphosphoryldolichol. This enzyme may be important in the regulation of secretion since the alpha-Glc-1-P present on the 62-kDa phosphoglycoprotein appears to be removed and then rapidly replaced in response to secretagogue. PMID- 2555364 TI - Identification of a 27-kDa protein with the properties of type II iodothyronine 5'-deiodinase in dibutyryl cyclic AMP-stimulated glial cells. AB - Type II iodothyronine 5'-deiodinase (5'D-II) catalyzes the intracellular conversion of thyroxine (T4) to 3,5,3'-triiodothyronine (T3), producing greater than 90% of the bioactive thyroid hormone in the cerebral cortex. In cultured glial cells, expression of this enzyme is cAMP dependent. Exploiting the cAMP dependent nature of this enzyme in these cells and utilizing N-bromoacetyl-L-3'- or 5'-[125I]thyroxine (BrAc[125I]T4) to affinity label cellular proteins, a 27 kDa protein with the properties of this enzyme was identified. Intact cells labeled with BrAc[125I]T4 showed three prominent radiolabeled bands of proteins of Mr 55,000, 27,000, and 18,000 (p55, p27, p18, respectively) which incorporated approximately 80% of the affinity label. All three affinity-labeled proteins were membrane associated. One protein (p27) increased 5-6-fold after treating the cells for 16 h with dibutyryl cAMP; maximal specific incorporation of affinity label into the stimulated p27 was approximately 2 pmol/mg of cell protein in intact cells. Alterations in the steady-state levels of 5'D-II resulted in parallel changes in the quantity of p27. In cell sonicates, the rate of enzyme inactivation by BrAcT4 equaled the rate of affinity label incorporation into stimulated p27, whereas p55 and p18 showed little or no specific dibutyryl cAMP stimulated labeling. Enzyme substrates T4 and 3,3'5'-triiodothyronine (rT3) specifically blocked p27 labeling, whereas T3 and the competitive 5'D-II inhibitor EMD 21388 (a synthetic flavonoid) were much less effective. Iopanoate, an inhibitor of all deiodinase isozymes, was ineffective in blocking p27 labeling. Inhibition kinetics revealed that iopanoate was a noncompetitive inhibitor of dibutyryl cAMP-stimulated glial cell 5'D-II, suggesting that it interacts at a site distant from the substrate-binding site. These data identify a cAMP-inducible membrane-associated protein (p27) that has many of the properties of 5'D-II. PMID- 2555365 TI - Drosophila melanogaster strand transferase. A protein that forms heteroduplex DNA in the absence of both ATP and single-strand DNA binding protein. AB - The purification of a Drosophila strand transfer protein is described, which involves Bio-Rex 70, Superose 6, Mono S, and single-stranded DNA-agarose chromatography. A 105,000-dalton polypeptide copurifies with the strand transfer activity on the last two column steps. The strand transferase carries out strand transfer at an unusually low protein:single-stranded DNA ratio and requires neither a nucleotide cofactor nor exogenous single-strand DNA binding protein to form heteroduplex DNA. Biochemical analysis of the reaction products has established that one strand of the DNA duplex is displaced during the reaction. Several properties, including the kinetics and stoichiometry of strand transfer, differentiate this activity from previously characterized strand transferases. PMID- 2555366 TI - Hydrodynamic, electron microscopic, and ligand-binding analysis of the Epstein Barr virus/C3dg receptor (CR2). AB - The interaction of the Epstein-Barr virus/45-kDa proteolytic fragment of C3 (C3dg) receptor (CR2) with its viral ligand, the Epstein-Barr virus glycoprotein gp350/220, initiates the sequence of events leading to virus internalization and B lymphocyte transformation. Soluble recombinant receptor (rCR2) and gp350/220 as well as the natural ligand, C3dg, were subjected to a number of analytical techniques including gel permeation chromatography, density gradient ultracentrifugation, circular dichroism, and electron microscopy in order to determine their hydrodynamic, structural, and binding properties. Both rCR2 and gp350/220 were found to be highly extended proteins (f/fo = 2.1 and 2.4/2.2, respectively). C3dg, in contrast to the viral ligand, is only somewhat elongated (f/fo = 1.5). Soluble rCR2, visualized by high resolution electron microscopy, was shown to be an extended, highly flexible molecule comprised of ringlet domains, each approximately 24.1 A in length, which likely correspond to the short consensus repeat motif deduced from the CR2 cDNA nucleotide sequence. Ligand-binding studies carried out under physiological conditions indicated that gp350/220 binding to rCR2 was saturable and univalent, with a dissociation constant of 3.2 nM. In contrast, monomeric C3dg did not bind to rCR2 under physiological conditions; however, at reduced ionic strength, monomeric C3dg binding could be measured. These studies indicate that the affinity of the C3dg monomer for rCR2 under physiologic conditions is approximately 10(4)-fold less than that of the viral ligand. The molecular properties of rCR2 revealed in these studies provide essential information for future studies of the biologic functions of the Epstein-Barr virus/C3dg receptor. PMID- 2555367 TI - 1,2-dioctanoyl-sn-glycerol can stimulate neutrophils by different mechanisms. Evidence for a pathway that does not involve phosphorylation of the 47-kDa protein. AB - Neutrophils treated with 1,2-dioctanoyl-sn-glycerol (DiC8) are known to release large quantities of superoxide (O2-) and to exhibit an intense phosphorylation of two proteins with molecular masses of approximately 47 and 49 kDa. In this paper, we report that O2- release from guinea pig cells stimulated with a near optimal amount of DiC8 (2.0 microM) is markedly inhibited (greater than or equal to 70%) by antagonists of protein kinase C (i.e. 150 nM staurosporine; 200 microM 1-(5 isoquinolinylsulfonyl)-2-methyl-piperazine (H-7], whereas that from cells stimulated with an optimal amount of DiC8 (7.8 microM) is not (approximately 25% inhibition). However, staurosporine (150 nM) effectively reduced the level of phosphorylation of the 47- and the 49-kDa proteins to that observed in unstimulated cells when either amount of DiC8 (i.e. 2.0 or 7.8 microM) was utilized. Thus, neutrophils stimulated with 7.8 microM DiC8 in the presence of staurosporine release large quantities of O2- without an enhanced phosphorylation of the 47- and the 49-kDa proteins. In contrast, these antagonists of protein kinase C effectively blocked O2- release from neutrophils stimulated with an optimal amount of phorbol 12-myristate 13-acetate (PMA), and the percentage of inhibition was not affected by increasing the concentration of PMA 160-fold. These data show that DiC8 and PMA, both activators of protein kinase C, can have distinct effects on O2- release by neutrophils. Moreover, they suggest that DiC8 (or a metabolite) under certain circumstances may function in a stimulatory pathway for O2- release that is independent of protein kinase C. Differences in the morphology of neutrophils stimulated with PMA and DiC8 are presented. Ancillary data on human neutrophils are also provided. PMID- 2555368 TI - Regulation of endothelial cell coagulant properties. Modulation of tissue factor, plasminogen activator inhibitors, and thrombomodulin by phorbol 12-myristate 13 acetate and tumor necrosis factor. AB - The procoagulant response of endothelium to many stimuli alters the expression of tissue factor, thrombomodulin, and plasminogen activator inhibitors (PAI) PAI-1 and PAI-2. The regulation of these proteins was examined in cultured human endothelial cells treated with phorbol myristate acetate (PMA) or tumor necrosis factor (TNF). Unstimulated cells contained approximately 670 PAI-1 and approximately 100 thrombomodulin mRNA molecules/cell, whereas tissue factor and PAI-2 mRNAs were not detectable. By 3-5 h, PMA or TNF induced both tissue factor and PAI-2 to approximately 150-420 mRNA molecules/cell and both mRNAs declined to basal levels within several hours; however, PAI-1 and thrombomodulin mRNA levels did not change. Nuclear runoff assays showed that PMA, TNF, or cycloheximide induced transcription of the tissue factor gene, whereas the genes for thrombomodulin, PAI-1, and PAI-2 apparently were transcribed at the same relative rate in the presence or absence of these agents. Treatment of cells with cycloheximide stabilized tissue factor and PAI-2 mRNAs and increased their induction by PMA or TNF. The synthesis of tissue factor, PAI-1, and PAI-2 proteins paralleled their mRNA levels. The effects of TNF were similar to those of PMA with one exception. In contrast to PMA, TNF reduced thrombomodulin activity approximately 80% with no change in thrombomodulin mRNA levels. Thus, PAI-2 may be induced by inhibiting mRNA degradation. Tissue factor can be induced by stimulating transcription and potentially by inhibiting mRNA degradation. Thrombomodulin can be repressed by a translational or posttranslational mechanism. PAI-1 was not regulated under the conditions studied. The different effects of PMA and TNF on thrombomodulin expression indicate that some effects of TNF are not mediated solely by protein kinase C. PMID- 2555369 TI - Thrombin stimulates the activities of multiple previously unidentified protein kinases in platelets. AB - We have used a renaturation method to search for previously unidentified protein kinases in human platelets. The method involves subjecting lysates to denaturing gel electrophoresis, transferring the proteins to blotting membranes, and treating the blotted proteins with guanidine. The guanidine is then removed to allow the proteins to renature, and the blots are overlaid with [gamma-32P]ATP. We have identified 14 electrophoretically distinct, serine/threonine-specific protein kinases. Eleven of the kinases clearly differ in molecular weight from all previously described platelet serine/threonine kinases. Ten of these novel kinases (PK220, PK200, PK170, PK150, PK64, PK60, PK56, PK52, PK48, and PK40) were found to possess markedly increased in vitro activity when isolated from thrombin stimulated platelets, presumably as a result of thrombin-stimulated covalent modification. Treatment of intact platelets with the calcium ionophore ionomycin and phorbol 12-myristate 13-acetate also increased the in vitro activity of these kinases. The agonist-stimulated kinases could be divided into three classes: 1) one kinase whose activity was increased by in vivo phorbol ester treatment but not by ionomycin (PK150); 2) two kinases whose activity was increased by ionomycin but not phorbol ester (PK48 and PK40); 3) seven kinases whose activity was markedly increased by combinations of phorbol ester and ionomycin, but not by either agent alone (PK220, PK200, PK170, PK64, PK60, PK56, and PK52). This third mode of regulation is what would be expected of enzymes that mediate the biological effects of inositide-mobilizing stimuli. PMID- 2555370 TI - Insertion of L1 elements into sites that can form non-B DNA. Interactions of non B DNA-forming sequences. AB - Three rat L1 element integration (target) sites chosen at random can adopt non-B DNA structures in vitro at normal bacterial superhelical densities. These target sites contain, respectively, short, mixed (AT)n tracts that we show can form one or more cruciforms, short (GT)n tracts, or polypurine:polypyrimidine regions. These sites share no sequence homology, and a non-B DNA structure appears to be the only feature common to them all. When the right end of the L1Rn3 element which forms a complex series of non-B DNA structures including two triplexes, and its target site which undergoes cruciform extrusion, are present on the same supercoiled molecule, they compete for available supercoil energy. The amount of non-B DNA formed at each site varies with pH, the concentration of cations, and the size of the topological domain. The implication of our findings for recombination of L1 elements and for the effect of these elements on contiguous DNA sequences is discussed. PMID- 2555371 TI - Nerve growth factor-induced decrease in the calpain activity of PC12 cells. AB - PC12 cells are a nerve growth factor-responsive clone derived from a rat pheochromocytoma. Treatment with nerve growth factor causes the cells to differentiate. One of the hallmarks of this differentiation is the generation of neurites. PC12 cells contain both calpain I and calpain II; about 90% of the total calpain activity is due to calpain II. Treatment of the cells with nerve growth factor causes a time-dependent decrease in calpain activity, more than 50% being lost over a 5-day period. Both the decrease in calpain activity and the growth of neurites are reversible upon the removal of nerve growth factor from the cultures. Agents other than nerve growth factor that cause neurite outgrowth, such as fibroblast growth factor and dibutyryl cyclic AMP, also cause a decrease in calpain activity. Calpain levels, as detected with immunoblotting or immunohistochemistry, show no decrease. Removal of calpastatin, the endogenous inhibitor of the calpains, by phenyl-Sepharose chromatography increases the calpain activity of extracts from both control and nerve growth factor-treated cells and brings the activity in the extracts from treated cells up to the activity in those from controls. Calpastatin-containing fractions from extracts of nerve growth factor-treated cells inhibit more calpain activity than do comparable fractions from control cells. These studies suggest that nerve growth factor causes a decrease in the activity of calpain in morphologically differentiating PC12 cells by causing an increase in the activity of calpastatin. PMID- 2555372 TI - Interosseous transfer of tibialis posterior for common peroneal nerve palsy. AB - The interosseous route remains popular for tibialis posterior tendon transfer for drop-foot. It leaves a smaller range of movement than the circumtibial route, but lengthening the calcaneal tendon may improve this. The results of this present series indicate that, in order to predict a good functional result, the ankle must be held in at least 20 degrees of dorsiflexion at the time of tendon transfer. PMID- 2555373 TI - Ganciclovir--a review of pharmacology, therapeutic efficacy and potential use for treatment of congenital cytomegalovirus infections. PMID- 2555374 TI - Separation of lipoxins and leukotrienes from human granulocytes by high performance liquid chromatography with a Radial-Pak cartridge after extraction with an octadecyl reversed-phase column. PMID- 2555375 TI - Simple apparatus for capillary zone electrophoresis and its application to protein analysis. AB - The construction of a simple apparatus for capillary zone electrophoresis is described, consisting of an optical system allowing direct absorbance measurement in the capillary in UV light, an evaluating electronic module and a high potential source. An attempt was made to achieve maximum sensitivity with a simple construction. In the electronics, care was taken to obtain a quiet baseline and to optimize the signal-to-noise ratio. That part of the noise which is of the frequency band of the signal is filtered off. Both suction and electrophoretic sample introduction are possible. According to operator's choice, the apparatus can be run under constant voltage or current and is protected against overloading. The high-potential electrode chamber contains separate buffer and sample compartments and its construction offers an easy interchange between the running and sampling positions. The applicability of the system to the separation of amino acids as phenylthiocarbamyl derivatives, peptides and both artificial and naturally occurring protein mixtures is demonstrated. PMID- 2555376 TI - Efficiency of isolation of human rotavirus in primary African green monkey kidney cells. AB - Out of 212 human rotavirus (HRV) containing fecal specimens, 173 (81.6%) yielded virus on first passage in primary African Green monkey kidney cells (AGMK), while additional 34 specimens, did not yield virus on first passage. However, following blind passages, 18 of the 34 yielded virus in passage levels 2-8, thus raising the overall isolation rate to 90.1%. The isolation rate of HRV strains obtained in embryonic Rhesus monkey kidney cell line (MA-104), was only 41.4%. ELISA tests performed on fluids from infected cell cultures proved to be an efficient tool to measure virus replication. No differences were encountered in the isolation rates between subgroup I and II strains, while viruses lacking the antigenic determinants of both subgroups did not grow at all. However, one of those unusual group A strains was isolated and grew well in AGMK cells. Primary AGMK and MA-104 cells supported the growth of tissue culture adapted virus most efficiently when compared with six human and primate cell types. PMID- 2555377 TI - An improved ELISA and serum neutralisation test for the detection of turkey rhinotracheitis virus antibodies. AB - The development of a double-well ELISA test for the detection of turkey rhinotracheitis (TRT) virus antibody in turkey and chicken sera, utilising a streptavidin-biotin detection protocol is described. This test was compared with a conventional ELISA, which detects antibody using a peroxidase-labelled anti turkey IgG. The double-well streptavidin-biotin ELISA was more sensitive than the conventional ELISA, and, due to the consistently low background values obtained, was further modified to a single-well ELISA test. Antibody titres obtained using the single-well streptavidin-biotin ELISA were similar to those obtained using the conventional ELISA. A microtitre serum neutralisation test for TRT virus read using an indirect immunoperoxidase (IIP) detection protocol after 48 h was also developed and compared to the standard neutralisation test employing a 7 day incubation and read by CPE. The specificity of the single-well streptavidin biotin ELISA was confirmed using the rapid (IIP) serum neutralisation test. PMID- 2555378 TI - Differential detection of viral DNA and RNA in situ in cells infected with human cytomegalovirus. AB - We have analyzed the ability to use in situ cytohybridization to distinguish between human cytomegalovirus (HCMV) DNA and RNA in human cells infected in vitro. Two different viral-specific probes were used, one for an abundantly expressed late gene, and one which includes at least two genes coding for immediate early (IE) proteins. In productively infected cells, hybridization of the late gene probe extended over both the nucleus and cytoplasm and was RNase sensitive, whereas hybridization of the IE probe was restricted to the nucleus and was DNase-sensitive. In nonproductively infected cells hybridization of the IE probe was localized to the cytoplasm and was RNase-sensitive. The specific nuclease sensitivities indicate that a cytoplasmic hybridization pattern correlates with detection of viral RNA sequences, whereas a nuclear pattern represents detection of viral DNA. These results demonstrate that in situ cytohybridization can potentially be used to determine the extent of HCMV infection in a particular tissue or cell type by distinguishing between transcription and replication of specific viral genes. PMID- 2555379 TI - Detection of human papillomavirus type 16 DNA in cervical swabs and formalin fixed, paraffin-embedded squamous cell carcinomas of non-genital tissues using a synthetic oligodeoxynucleotide probe. AB - The potential of using a chemically synthesized oligodeoxynucleotide as a diagnostic probe to detect human papillomavirus type 16 (HPV-16) in genital infections was evaluated by comparing it with a cloned full-length HPV-16 probe in dot-blot DNA hybridizations. An oligonucleotide sequence, 20 bases in length from the E6 region of HPV-16 (E6 oligo) and different from the DNA sequences of HPV types 6, 11 and 18 by at least 2 base pairs, was chosen for chemical synthesis. The oligoprobe, which was 5'-end labelled with [32P]dATP, was found to be specific, but approximately ten times less sensitive than the full-length radiolabelled probe of HPV-16, in dot-blot hybridizations with the DNA of HPV-6, 11, -16 and -18, HPV positive and negative cell-lines. From 36 cervical or vulval scrapes two samples were found positive with both cloned HPV-16 and oligoprobe hybridization. Of 21 samples of formalin-fixed, paraffin-embedded squamous cell carcinomas originating from anus, oesophagus, penis, colon, breast and skin only 4 anal squamous cell carcinomas were positives when hybridized with cloned HPV-16 DNA or with the oligoprobe. This study confirms that HPV-16, which is frequently associated with squamous cell carcinoma of the cervix is also strongly associated with squamous cell carcinoma of the anus. PMID- 2555380 TI - Comparison of an antibody-capture IgM enzyme-linked immunosorbent assay with IgM indirect immunofluorescence for the diagnosis of acute Sindbis and West Nile infections. AB - The development and evaluation of an antibody-capture ELISA for the detection of IgM antibodies to Sindbis (SIN) and West Nile (WN) viruses are described. Comparison of the ELISA results with those obtained by indirect immunofluorescence (IIF) antibody tests using both fluorescein and biotinylated anti-human IgM conjugates, showed that the former technique was both more sensitive and specific than the IIF methods. There were no false positives by the ELISA whereas with the IgM-IIF assays a high percentage of false positives were obtained. These were due to rheumatoid factor and also to an interfering factor which was not detected by the RF latex agglutination test. Absorption of the sera with anti-IgG was necessary to eliminate this interference in the IgM-IIF tests. PMID- 2555381 TI - Predictive factors for initial cure and relapse rate after pituitary surgery for Cushing's disease. AB - Twenty-seven patients with Cushing's disease underwent transsphenoidal pituitary surgery. After the operation 16 patients were cured, and 11 remained hypercortisolemic. In the cured patients a significantly lower incidence of paradoxical responsiveness to TRH or to LRH was found preoperatively (1 of 16) than in the failures (6 of 11; P less than 0.02, by Fisher's chi 2 test). Furthermore, responsiveness of cortisol to CRH administration was significantly lower in the failures [maximum, 0.30 +/- 0.10 mumol/L (10.8 +/- 3.6 micrograms/dL) vs. 0.50 +/- 0.07 mumol/L (18.1 +/- 2.5 micrograms/dL) in the cured patients; P less than 0.05]. There were no differences in basal plasma cortisol levels, ACTH levels, or suppressibility by dexamethasone between the 2 groups. In the 16 patients who were cured initially, 4 patients relapsed after a mean period of 4 yr. These 4 patients had significantly higher basal cortisol levels postoperatively than those who remained in remission [0.14 +/- 0.03 mumol/L (5.1 +/- 0.8 micrograms/dL) vs. 0.04 +/- 0.01 mumol/L (1.4 +/- 0.3 micrograms/dL); P less than 0.01]. Cortisol responses to CRH after the operation positively correlated with the basal cortisol levels at that time (P less than 0.05, by Spearman's rank correlation test; r = 0.64), thus, the relapsing patients had higher cortisol responses to CRH than patients who stayed in remission [maximum response, 0.31 +/- 0.07 (11.2 +/- 2.5 micrograms/dL) vs. 0.12 +/- 0.03 mumol/L (4.3 +/- 1.1 microgram/dL), respectively; P less than 0.05]. We conclude that 1) patients responding paradoxically to TRH and/or LRH have a lower chance of being cured after pituitary surgery; and 2) patients with higher cortisol levels (greater than 0.10 mumol/L; 3.6 micrograms/dL) after being cured initially have a higher chance of recurrence of their disease. PMID- 2555382 TI - Molecular characterization of a Leydig cell tumor presenting as congenital adrenal hyperplasia. AB - We present an unusual patient with a Leydig cell tumor to show that greatly elevated serum concentrations of 17-hydroxyprogesterone (17OHP) may not be diagnostic of congenital adrenal hyperplasia (CAH). A 3.5-yr-old boy had a small testicular mass and plasma 17OHP concentrations of 147-333 nmol/L (4,850-11,000 ng/dL), suggesting CAH with adrenal rests. However, normal plasma cortisol values and the unresponsiveness of the 17OHP concentration to dexamethasone suppression or ACTH stimulation suggested a diagnosis of Leydig cell tumor. A 4-fold elevation in plasma 21-deoxycortisol compared with a 200-fold elevation in 17OHP suggested that the elevated 17OHP derived from the normal pathway of testosterone synthesis in the testis. This was proven by normalization of all hormonal values after tumor resection. Compared to the abundance of mRNA for P450c17, the tumor contained unusually large amounts of mRNA for P450scc, the cholesterol side-chain cleavage enzyme, which is the rate-limiting step in steroid hormone synthesis. Increased P450scc activity, which increased the conversion of cholesterol to pregnenolone, apparently permitted the 17,20-lyase activity of P450c17 to become rate limiting, thus accounting for the increased secretion of 17OHP. Thus, Leydig cell tumors can produce quantities of 17OHP previously reported only in CAH due to 21-hydroxylase deficiency. The molecular characterization of steroidogenic mRNAs in this tumor indicates an unusual ratio in the expression of the genes for the steroidogenic enzymes, probably accounting for the unusual pattern of serum steroids. PMID- 2555383 TI - Relationships between histomorphometric features of bone formation and bone cell characteristics in vitro in renal osteodystrophy. AB - To determine whether abnormal bone cell recruitment or differentiation may be involved in the development of aplastic bone lesion in renal osteodystrophy we have compared histomorphometric parameters of bone formation and in vitro behavior of osteoblastic cells isolated from the trabecular bone surfaces in 37 dialysis patients with osteitis fibrosa, normal bone formation rate, or aplastic bone lesion. The bone cell responses to human PTH-(1-34) (20 nmol/L), as evaluated by intracellular cAMP production, and to 1,25-dihydroxyvitamin D (10 nmol/L), as assessed by osteocalcin synthesis, were not different from normal in patients with low, normal, or high bone formation rates. Osteoblastic cells isolated from patients with a high bone formation rate and markedly elevated serum iPTH and osteocalcin values had a higher than normal DNA replication in primary culture. The peak of [3H]thymidine incorporation, the maximal DNA synthesis, and the area under the growth curve were 4.4- to 6.3-fold increased in osteitis fibrosa compared to those in normal bone cells obtained from age-matched individuals. By contrast, [3H]thymidine incorporation in bone cells from aplastic patients was about 25% of normal and only 5% of the value in osteitis fibrosa. The decreased DNA replication of cultured bone cells in aplastic patients was unrelated to trabecular bone aluminum staining, but was associated with low serum immunoreactive PTH values compared to those in other groups of patients. These results show that high bone formation in uremic osteoitis fibrosa is associated with higher than normal [3H]thymidine incorporation in bone cells in vitro, whereas low bone formation in aplastic patients results from lower than normal DNA replication and suggest that the defective osteoblastic recruitment in aplastic patients may be related to factors other than aluminum, including inappropriate PTH secretion. PMID- 2555384 TI - Regulation of insulin-like growth factor-binding protein-1 production in human granulosa-luteal cells. AB - Human follicular fluid contains the insulin-like growth factor-binding protein (IGFBP-1) synthesized by ovarian granulosa cells. We studied the regulation of IGFBP-1 production by the granulosa-luteal cells from the hyperstimulated follicles of patients attending an in vitro fertilization program. The cells were first allowed to attach and recover from the hyperstimulation for 2 days. Then, a protein kinase-C activator, 12-O-tetradecanoyl phorbol-13-acetate (TPA), and adenylate cyclase activators, gonadotropins, FSH, hCG, cholera toxin, or prostaglandin E2 (PGE2) were added to the cells. The gonadotropins failed to increase IGFBP-1 production, whereas it was enhanced by TPA and to a lesser extent by cholera toxin and PGE2. The maximal response to TPA occurred at the concentration of 1.0 ng/mL, and the enhancing effect of TPA was detected at 24 h. PGE2 stimulated IGFBP-1 production; the lowest effective concentration was 10(-8) mol/L. The mean highest response was 4.3-fold and occurred at the PGE2 concentration of 10(-5) mol/L. The effect of PGE2 on IGFBP-1 production became detectable at 24 h, and it continued to increase up to 72 h. PGE2 also increased granulosa-luteal cell progesterone production in a dose- and time-dependent manner. Incorporation of [35S]methionine into immunoreactive IGFBP-1, as detected by sodium dodecyl sulfate-polyacrylamide electrophoresis and fluorography, was also increased by TPA. This suggests that TPA accelerated the synthesis of IGFBP 1. Our results indicate that the production of IGFBP-1 by human granulosa-luteal cells can be regulated both via protein kinase-C- and adenylate cyclase-dependent pathways. PMID- 2555385 TI - Differences in mineral metabolism among nonhuman primates receiving diets with only vitamin D3 or only vitamin D2. AB - We tested for differences in aspects of mineral metabolism during the administration of diets with only vitamin D3 or only vitamin D2 in four nonhuman anthropoid primate species [two catarrhini, Macaca fascicularis (crab-eating macaque) and Macaca mulatta (rhesus macaque), and two platyrrhini, Saimiri sciureus (squirrel monkey) and Aotus vociferans (night monkey)]. All four species maintained approximately 2- to 3-fold higher serum 25-hydroxyvitamin D (25OHD) level while receiving vitamin D3 than while receiving similar amounts of vitamin D2. Serum 25OHD in M. mulatta receiving the standard primate dietary supplement of vitamin D3 was high enough (360 +/- 60 vs. 70 +/- 25 nM in vitamin D supplemented humans; P less than 0.0001) to suggest that this widely used level of vitamin D3 supplementation is excessive for some M. mulatta. Serum 24,25 dihydroxyvitamin D [24,25-(OH)2D] in A. vociferans was uniquely high [P less than 0.01; species mean, 19 +/- 5, 95 +/- 12, and 27 +/- 5 nM in groups receiving diets with 1.5 IU vitamin D3/g, 6.6 IU vitamin D3/g, and 15 IU vitamin D2/g, respectively; mean 24,25-(OH)2D from the other three species pooled across three diets was 7 +/- 5 nM]. We confirmed relative resistance to 1,25-(OH)2D in S. sciureus, manifested by osteomalacia and moderately high serum 1,25-(OH)2D. Serum 1,25-(OH)2D in S. sciureus increased 4-fold (P less than 0.05) when the precursor in serum was changed from 250HD3 to 250HD2, suggesting that this species shows more severe resistance to 1,25-(OH)2D2 than to 1,25-(OH)2D3. In conclusion, we found many differences in vitamin D metabolism among four nonhuman anthropoid primate species. The striking feature in A. vociferans (high, 24,25-(OH)2D without high 25OHD in serum independent of whether diet contained only vitamin D3 or only vitamin D2) should allow determination of whether 24,25-(OH)2D functions as a unique agonist or an inactive metabolite in this species. PMID- 2555386 TI - Molecular forms of serum insulin-like growth factor (IGF)-binding proteins in man: relationships with growth hormone and IGFs and physiological significance. AB - Insulin-like growth factor-I (IGF-I) and IGF-II are associated in the blood with specific binding proteins (BPs), forming complexes that elute in gel filtration with estimated mol wt around 40 and 150 kD. The latter appears to be under GH control. Five molecular forms of BP (41.5, 38.5, 34, 30, and 24 kD) have been identified by Western blotting using 125I-labeled IGF. All five forms are present in the smaller complexes, but only the 41.5- and 38.5-kD forms are found in the larger complexes. In this study immunoblotting showed that the 41.5- and 38.5-kD forms were recognized by antibodies directed against the GH-dependent BP purified from human plasma, and the 30-kD form was recognized by antibodies directed against the BP purified from amniotic fluid. The 34- and 24-kD forms proved to be immunologically unrelated to the other three. In sera with large quantities of the 41.5- and 38.5-kD forms, an additional band was often observed immediately ahead of the migration front of the 30 kD band. This was recognized by the anti GH-dependent BP antibody and probably corresponds to a degradation product of the 41.5- and 38.5-kD BPs. Serum 41.5- and 38.5-kD BPs have been found to be elevated in acromegaly, where GH hypersecretion causes increased IGF-I levels, and diminished in cases of genetic or idiopathic GH deficiency and defects of the GH receptor (Laron's syndrome), where both IGF-I and IGF-II are decreased, as well as in Pygmy adults and children who have isolated IGF-I deficiency. In all of these conditions, the proportions of the 34- and 30-kD forms were inversely related to those of the 41.5- and 38.5-forms. Under treatment, the BP profiles tended to return to normal. In cases of GH deficiency caused by a tumor, the BP profiles resembled those of hypopituitary or normal serum, depending on whether IGF levels were diminished or normal. It, therefore, seems that BP synthesis is coordinated with IGF-I synthesis and may not be directly GH dependent. The results of neutral pH gel filtration analysis of hypopituitary (idiopathic and tumoral) and normal sera point to a relationship between the levels of circulating IGFs and those of the 150-kD IGF-BP complex whose binding units are the 41.5- and 38.5-kD BPs. It, therefore, seems that the 150-kD complex controls the bioavailability of IGF-I and IGF-II.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2555387 TI - Failure of bronchial lavage to detect elevated levels of adrenocorticotropin (ACTH) in patients with ACTH-producing bronchial carcinoids. AB - Saline lavage of the major bronchial segments was performed in five patients immediately preceding lobectomy to remove an ACTH-producing bronchial carcinoid. Lavage fluid from each bronchus was concentrated, and ACTH determinations were performed. Elevated concentrations of ACTH were not demonstrated in the aspirate from the bronchus containing the known tumor. Two additional patients with ectopic ACTH syndrome from an unknown primary source also underwent selective segmental bronchial lavage for ACTH determination. Neither patient demonstrated ACTH gradients in the lavage specimens. One was found to have an ACTH-producing bronchial carcinoid 18 months after negative lavage. Selective segmental bronchoscopic lavage with measurement of ACTH levels on the aspirate is not an effective technique for detecting ACTH-producing bronchial carcinoid tumor. PMID- 2555388 TI - Plasma pancreastatin-like immunoreactivity in various diseases. AB - Plasma pancreastatin (PST)-like immunoreactivity in normal subjects and patients with various diseases was estimated by a RIA, using antiserum raised against a synthetic C-terminal peptide of human PST deduced from the sequence of human chromogranin-A. The mean level +/- SEM was 13.2 +/- 0.6 pmol/L in normal subjects, but was significantly higher in patients with chronic renal failure (526.7 +/- 48.5). An immunoreactive form corresponding to a human PST-like sequence [human chromogranin-A-(250-301)] and a larger form were detected by gel filtration of plasma from these patients, suggesting accumulation of the larger molecular form in these patients. A significant increase in PST-like immunoreactivity was also found in patients with liver cirrhosis (20.8 +/- 3.0 pmol/L), but not in patients with noninsulin-dependent diabetes mellitus, chronic pancreatitis, or pancreatic cancer. Elevated levels were found in 16 of the 21 patients with small cell lung carcinoma examined. High levels were also found in 3 of 11 patients with islet cell tumor. PMID- 2555389 TI - Establishment of titration system for human herpesvirus 6 and evaluation of neutralizing antibody response to the virus. AB - The susceptibilities of seven T-cell lines to human herpesvirus 6 (HHV-6) infection were examined. MT-4 cells were the most susceptible of these lines to infection with this virus. Therefore, chemically adhered MT-4 cell monolayers were used for infectious HHV-6 assay by indirect immunofluorescent-antibody (IFA) staining. When cell monolayers were fixed 30 to 45 h postinfection, the foci stained with IFA were easy to count and a linear relationship was observed between the number of foci and the virus concentration. MT-4 cell monolayers were also used for a focus reduction neutralizing-antibody test. In this test, sera from patients in the convalescent stage of exanthem subitum all showed significant neutralizing activity (1:80 to 1:320), whereas sera from patients in the acute stage of disease showed no detectable neutralizing activity. The titers of neutralizing antibody correlated well with the levels of anti-HHV-6 antibodies detected by IFA. PMID- 2555390 TI - Identification of pseudorabies virus-exposed swine with a gI glycoprotein enzyme linked immunosorbent assay. AB - A monoclonal antibody specific for the gI glycoprotein of virulent pseudorabies virus was produced and used to affinity purify gI glycoprotein. The purified gI was used in an enzyme-linked immunosorbent assay (ELISA) that identified and differentiated field virus-exposed animals from animals vaccinated with gI deleted virus. The gI ELISA was evaluated by comparing it with the virus neutralization test and with a standard ELISA which does not distinguish between vaccinated and naturally infected animals. Pigs vaccinated with a gI-deleted vaccine were seropositive by the virus neutralization or standard ELISA but were seronegative in the gI ELISA. Nonvaccinated and vaccinated animals were detected as seropositive in the gI ELISA only after exposure to gI-containing field virus. Exposed animals were detected as early as day 7 and for as long as 141 days after field virus exposure. As little as 10(2.7) PFU of field virus was sufficient to seroconvert negative animals in the gI ELISA. Pseudorabies virus-seronegative animals which received multiple doses of gI-deleted vaccine remained seronegative in the gI ELISA. The use of this test to monitor swine for pseudorabies virus infection would offer significant benefits towards eradication of the disease. PMID- 2555391 TI - Enumeration of viral antigen-reactive helper T lymphocytes in human peripheral blood by limiting dilution for analysis of viral antigen-reactive T-cell pools in virus-seropositive and virus-seronegative individuals. AB - A limiting-dilution analysis technique was developed which enumerates human T cells with the capacity to secrete T-cell growth factors such as interleukin 2 after contact with herpes simplex virus type 1 (HSV-1) or cytomegalovirus (CMV) antigens (operationally defined as virus-reactive helper T cells [HTL]). By using this limiting-dilution analysis technique, the peripheral blood of HSV seropositive individuals was analyzed for the frequency of HSV antigen-reactive HTL and for the ability either to proliferate or to secrete detectable T-cell growth factors in conventional HSV antigen-stimulated lymphocyte cultures. We found that the magnitudes of the latter two responses did not correlate directly with the frequency estimates of HSV antigen-reactive HTL. The study was expanded to analyze both HSV and CMV reactivities within individuals. Those who were seropositive for HSV or CMV were found to have relatively high HTL frequencies for the viral antigens to which they were sensitized. However, those who were seronegative for one of the viruses often had HTL reactive with that virus in their peripheral blood. These latter HTL frequencies were highly variable and ranged from undetectable to quite prominent, even within the same individual at different times. In general, it was found that viral antigen-reactive serologic activity does not necessarily reflect the status of viral antigen-reactive cell mediated immunity in humans and that viral antigen-induced T-cell responses may be unexpectedly complex, rather than absent, in individuals who are seronegative for a particular virus. PMID- 2555392 TI - Antibody response to recombinant lambda gt11 fusion proteins in cytomegalovirus infection. AB - Significant epitopes of two of the major cytomegalovirus antigens, a nonstructural DNA-binding protein of 52 kilodaltons (kDa) and a structural phosphoprotein of 150 kDa, expressed as fusion proteins with the beta galactosidase, were induced in Escherichia coli after infection with recombinant lambda gt11 clones. The epitopes were then used in immunoblotting to assay specific immunoglobulin G (IgG) and IgM in several groups of sera from long-term seropositive subjects and from patients undergoing primary or secondary virus infection. The data obtained showed that IgM reacting with the 52-kDa nonstructural antigen are linked to primary virus infection and can therefore be considered a serological marker of this infection. PMID- 2555393 TI - Human herpesvirus 6 infection and Kawasaki disease. AB - Eighteen of a total of 22 serum specimens (81.8%) from patients with Kawasaki disease were positive for immunoglobulin G or M antibodies to human herpesvirus 6, whereas 10 of 16 age- and sex-matched healthy controls (62.5%) were seropositive. Additionally, increased geometric mean antibody titers of immunoglobulin G were shown in these patients. These results suggest that the status of human herpesvirus 6 infection may be a reflection of the immunologic alterations that are associated with Kawasaki disease. PMID- 2555394 TI - Rapid screening test for the diagnosis of rotavirus infection. AB - The early diagnosis of human rotavirus infection is essential for effective patient management and infection control. We report here a rapid, easy-to perform, and inexpensive test for rotavirus detection. The viral RNA is extracted directly from the stools and electrophoresed on 1% agarose gels. Currently available immunoassays for routine diagnostic purposes are directed at the common group A-specific antigen. As reports become available on human gastroenteritis caused by the atypical or novel rotaviruses, this technique presents an added advantage in that it can detect both group A and non-group A rotaviruses. PMID- 2555395 TI - Tumor necrosis factor-alpha enhances interferon-gamma-mediated class II antigen expression on astrocytes. AB - Astrocytes can function as antigen-presenting cells (APC) upon expression of class II antigens, which are induced by interferon-gamma (IFN-gamma). Tumor necrosis factor-alpha (TNF-alpha) can act synergistically with IFN-gamma with respect to class II expression on a variety of cells. As brain cells themselves can secrete TNF-like factors upon stimulation, we examined the effect of TNF alpha on IFN-gamma-mediated class II induction on astrocytes. TNF-alpha alone had no effect on class II expression, but did synergize with IFN-gamma for enhanced expression of class II antigens. The specificity of TNF-alpha activity was demonstrated by blocking the amplifying effect of TNF-alpha with a polyclonal anti-TNF-alpha antibody. Kinetic analysis of the synergistic effect indicated that optimal TNF-alpha enhancement of class II expression was observed when astrocytes were pretreated with IFN-gamma 12-24 h prior to TNF-alpha addition. A possible mechanism for the synergistic action between IFN-gamma and TNF-alpha may be increased TNF-alpha receptor expression by IFN-gamma. Astrocytes treated with IFN-gamma for 24 h express more TNF-alpha receptors (3900/cell) than do untreated astrocytes (2483/cell), with no significant change in the binding affinity (Kd). These results suggest that the synergistic activity of TNF-alpha requires an inductive signal from IFN-gamma, which in part may be increased TNF-alpha receptor expression. Altogether, our observations indicate that TNF-alpha enhances ongoing class II major histocompatibility complex gene expression in rat astrocytes, which in this system is initially induced by IFN-gamma. TNF-alpha exerts its effect by binding to high affinity TNF-alpha receptors on astrocytes, whose expression is also enhanced by IFN-gamma. These two cytokines work in concert to elevate class II expression on astrocytes, an event which can contribute to initiation and/or perpetuation of intracerebral immune responses. PMID- 2555396 TI - Morphometric and flow cytometric analysis of small cell undifferentiated carcinoma of the bladder. AB - Eighteen cases of primary small cell carcinoma of the bladder were studied. Three patients survived for two years and one survived for five years, which was significantly worse when compared with poorly differentiated transitional cell carcinoma (WHO grade 3). Aggressive tumour behaviour was independent of the presence of neuroendocrine characteristics. Morphometric analysis showed that the nuclear size, which was comparable with that reported in pulmonary small cell carcinoma, was significantly smaller than in poorly differentiated transitional cell carcinoma. Nuclear morphometry may therefore help identify small cell bladder carcinoma, especially in the absence of neuroendocrine differentiation characteristics. DNA flow cytometry of paraffin wax embedded specimens showed aneuploidy in 14 tumours--five were peritetraploid and two multiple aneuploid- and only three were diploid. Aggressiveness of small cell bladder carcinoma usually coincides with aneuploidy in most cases, but diploid tumours may also follow a rapid, lethal clinical course. PMID- 2555397 TI - Cytomegalovirus infection in gastrointestinal tracts of patients infected with HIV-1 or AIDS. AB - All gastrointestinal tract biopsy specimens from 190 patients positive for HIV-1 or with AIDS were reviewed to assess the prevalence of cytomegalovirus (CMV) infection, morphology of infected cells, and the associated histopathological features. Eighteen patients (10 (7.7%) of 129 HIV antibody positive and eight (13.1%) of 61 with AIDS) had CMV identified in 35 biopsy specimens from the following sites: oesophagus (n = 3); stomach (n = 6); small intestine (n = 4); colorectum (n = 18) and perianal area (n = 4). Eleven patients had CMV alone as the potential cause of symptoms and in seven there were coexistent pathogens or Kaposi's sarcoma. The appearance and type of infected cells at different sites was highly variable. Immunocytochemical techniques and electron microscopic examination were performed to confirm the presence of CMV antigen and CMV virus particles and to exclude the possibility of an adenovirus producing similar cytopathic changes. It is important to recognise the different morphological forms of infected cells, and the use of immunocytochemical techniques is recommended in patients at risk for CMV or in whom CMV infection is suspected. PMID- 2555399 TI - Serum angiotensin converting enzyme: a possible marker in Lennert's lymphoma? PMID- 2555398 TI - Antibodies to cytomegalovirus in renal allograft recipients: correlation with isolation of virus. AB - A cohort of 47 renal transplant recipients was studied prospectively for up to one year after transplantation. Cytomegalovirus (CMV) was isolated from 21 of the patients. The first time the virus was isolated seven patients were IgM positive, nine showed a significant rise in IgG titres, and 12 had a four-fold or greater rise in complement fixation titre. There was no significant difference in the time at which virus was first detected following transplantation between patients with primary CMV infection and those with reinfection or recurrent infection. In general, patients with primary infection shed virus consistently over long periods. Those with reinfection or recurrent infection shed virus intermittently or not at all. There were considerable differences between individual patients in the timing and pattern of the immune response. Taken overall, a four-fold rise detected by the complement fixation test correlated best with the onset of CMV shedding in primary infection. There was more variation in the pattern of antibody response in cases of reinfection or recurrent infection, with no single serological test correlating better than the others. It is concluded that serology is of limited value in the detection of active CMV infection after renal transplantation. PMID- 2555400 TI - Pulmonary adenoma: a variant of sclerosing haemangioma of lung? PMID- 2555401 TI - Studies on the pathology of Marek's disease following challenge in chicks vaccinated with three different vaccines. AB - The pathology of Marek's disease (MD) was studied in chickens vaccinated separately with three different vaccines, namely herpesvirus of turkeys (HVT), inactivated Marek's disease virus (IMDV) and a bivalent vaccine. The MD-specific mortality following challenge was significantly lower (2.0 per cent) in bivalent vaccinated birds than in those vaccinated with either HVT (5.9 per cent) or IMDV (13.7 per cent) alone. The occurrence, extent and severity of gross lymphomas and lymphoproliferative lesions in visceral organs was lower in the bivalent vaccinated birds than in other groups. Atrophy and degenerative changes were seen in unvaccinated and challenged birds but not in challenged and bivalent vaccinated birds. Mild degenerative lesions and slight RE cell hyperplasia occurred in HVT- and IMDV-inoculated and challenged birds. The low intensity and frequency of gross and histological lesions in bivalent-vaccinated birds following challenge is highly suggestive of better vaccine-induced resistance. PMID- 2555403 TI - Use of itraconazole in a patient with chronic mucocutaneous candidiasis. PMID- 2555402 TI - Dermatologic complications associated with administration of 2',3' dideoxycytidine in patients with human immunodeficiency virus infection. AB - We describe a distinctive mucocutaneous eruption that occurred in 14 of 20 (70%) patients with human immunodeficiency virus infection while they were being treated with a new therapeutic agent, 2'3'-dideoxycytidine. A maculopapular eruption developed in these patients on day 10 or 11 of treatment. Seven of 14 patients, especially those receiving higher-dose therapy, also had systemic symptoms. In addition, oral ulcers developed in 9 of 14 patients on days 4 to 6 of therapy. The occurrence of the cutaneous and oral lesions correlated with dose, route, and schedule of administration of 2'3'-dideoxycytidine. In most instances the mucocutaneous lesions resolved even with continuation of therapy. PMID- 2555404 TI - [The possibility of the germination of spores of pathogenic clostridia and bacilli in soil]. AB - A possibility of germination of clostridia (Cl. tetani and Cl. perfringens) and bacilli (Bac. anthracis, STI vaccine strain) has been studied in model experiments with native soil. Mature spores did not germinate upon contact with native soil of deferent agrochemical types. Addition of meat-pepton medium and other protein, amino acid, and sugar-containing media led only to "swelling" of spores. The data obtained support the conclusions drawn by many researches that pathogenic clostridia and bacilli do not germinate in soil. PMID- 2555405 TI - New bacteriophage vectors for the large scale production of single stranded insert DNA. AB - SSEV 18 and SSEV 19, derivatives of the bacteriophages M13mp18/19, are new versatile cloning vectors allowing the large scale preparation of single stranded (ss) insert DNA. Replacing the original multiple cloning site by a synthetic 96 bp DNA fragment, a new polylinker region has been introduced containing complementary sequences designed to form a stem structure where single stranded insert fragments can be excised via a 'master restriction' site. The usefulness of such a vector has been demonstrated by the cloning of a 900 bp HindIII fragment derived from the Mycoplasma hyorhinis 23 S rRNA gene. After excision the single stranded insert was labeled isotopically and tested for sensitivity and specificity in detecting homologous sequences in pure DNA or cellular material immobilized on filters. PMID- 2555406 TI - Mossbauer and ESR study of iron in livex--a product of blood. PMID- 2555407 TI - A microtiter plate assay for inorganic phosphate. AB - A microtiter assay for the detection of picomolar quantities of inorganic phosphate has been described. The assay, linear between 50 and 1000 pmol of inorganic phosphate, is simple and rapid, with results obtainable in several minutes. Results from 5'-nucleotidase and (Ca2+ + Mg2+)ATPase assays using this method were compared with conventional phosphate assays and showed a high degree of correlation. The high sensitivity of this assay and the small sample size needed allows its widespread use in biochemical studies involving the generation of inorganic phosphate. PMID- 2555408 TI - Local chemotherapy for rectal polyposis: intraluminal administration of 5 fluorouracil for postoperative control of adenomas in the retained rectum in familial polyposis. AB - We administered the anticancer drug 5-fluorouracil locally to treat rectal adenomas in four patients with familial adenomatous polyposis who had undergone total colectomy and ileorectal anastomosis. The drug was administered either in suppository form or in high concentration as a rectal enema. One patient was treated successfully with 5-fluorouracil suppositories without any side effects, but the suppository regimen had to be stopped in three other patients because of severe rectal urgency. However, one of these patients was treated effectively with rectal administration of a high-dose 5-fluorouracil solution. Adenomas near the anastomotic site or dentate line were difficult to treat. Although severe urgency was seen in most patients, we think that the intraluminal administration of anticancer agents may prove effective in treating polyposis of the rectum retained after colectomy in familial polyposis. It deserves further study. PMID- 2555409 TI - Acute hemoperitoneum associated with a tumor-like appearance of the liver in a cirrhotic woman. PMID- 2555410 TI - Detection of diaminobenzidine reactions using scanning laser confocal reflectance microscopy. AB - We used scanning laser confocal microscopy to visualize sites of peroxidatic activity as detected by the diaminobenzidine (DAB) reaction. Imaging was achieved by employing the reflectance mode of this instrument. Intense reflectance was detected after DAB localization of endogenous granule-associated myeloperoxidase in neutrophils and of the exogenous tracer horseradish peroxidase in mouse oocytes. Detection of DAB reaction products with confocal reflectance microscopy will probably be an important addition to the utility of this cytochemical technique. PMID- 2555411 TI - Endothelin-induced contraction of human peripheral resistance vessels is partly dependent on stimulation of sodium-hydrogen exchange. AB - The role of Na-H exchange in endothelin-induced contraction of human peripheral resistance vessels was investigated. Endothelin produced a dose-dependent contraction which was greatly attenuated in the presence of a low extracellular sodium concentration. Inhibition of Na-H exchange by the amiloride analogue 5 (N,N-hexamethylene) amiloride (5-NNHA) resulted in a greater than 65% relaxation of a maximal endothelin-induced contraction in the presence of normal extracellular sodium. However, in the presence of a low extracellular sodium concentration, inhibition of Na-H exchange only resulted in a 25% relaxation. These data suggest that endothelin-induced vasoconstriction of human peripheral resistance vessels is mediated in part by stimulation of Na-H exchange. PMID- 2555412 TI - Human IL-7: a novel T cell growth factor. AB - IL-7 is a hemopoietic growth factor that induces the proliferation of early B lineage cells. In the course of studies to determine its effect on human bone marrow cells, we noted a marked outgrowth of mature T cells. When T cells from the circulation were cultured with IL-7, a dose-dependent proliferative response was observed. The target cells included both the CD4+ and CD8+ subpopulations of T cells, but the memory T cells (CD45R-) were better responders than unprimed T cells (CD45R+). IL-7 induced the expression of receptors for IL-2 and transferrin and higher levels of the 4F2 activation Ag. Although T cell responses to suboptimal concentrations of IL-7 were enhanced by the addition of IL-2, the proliferative response to IL-7 was not inhibited by neutralizing antibody to the IL-2R (Tac), nor was IL-2 secretion detected in this response. This response pattern of mature T cells suggests an important role for IL-7 in normal T cell physiology in humans. PMID- 2555413 TI - Cholera toxin inhibits resting human T cell activation via a cAMP-independent pathway. AB - The catalytic subunit of cholera toxin (CT) can chemically modify the alpha polypeptides of certain G-binding proteins and thus alter their function. In order to study the involvement of CT-sensitive G proteins in T cell activation, we have utilized CT in an in vitro system in which purified, resting human peripheral T cells are activated by anti-CD3 antibodies and rIL-2. Perturbation of the TCR/CD3 molecular complex by anti-CD3 antibodies causes changes in membrane phospholipids and induces a rise in cytoplasmic Ca2+. These events, however, are insufficient to allow progression into cellular proliferation and addition of IL-2 is required. Under these conditions, treatment of cells with a low concentration of CT (2 ng/ml) causes a significant inhibition of the anti-CD3 induced calcium event as well as the anti-CD3 plus IL-2-stimulated proliferation. Under our experimental conditions, inhibition of both proliferation and intracellular Ca2+ elevation by CT requires the involvement of the TCR/CD3 complex. This is supported by the observation that the toxin does not inhibit either the proliferation triggered by ionomycin and PMA or the Ca2+ influx induced by the ionophore. These data suggest that in TCR/CD3-mediated T cell activation CT acts at a point between TCR/CD3 perturbation and the generation of intracellular Ca2+. In view of the ability of CT to activate the alpha subunit of the G protein that stimulates adenyl cyclase (G alpha s), it is possible that the effect of CT on T cells is secondary to intracellular elevation of cAMP. However, measurement of cAMP levels both early after CT addition and at later time points, when proliferation is maximal, reveals lack of cyclic nucleotide accumulation. The presented data are consistent with the interpretation that the CT-mediated inhibition is caused by the modification of a G-binding protein that is either directly or indirectly associated with triggering of T cells via the TCR/CD3 molecular complex. The data also suggest that this protein is not G alpha s and it probably represents an as yet unidentified moiety or one of the several G proteins that have been recently described as regulators of phospholipase C activation. PMID- 2555414 TI - Release of platelet-activating factor and the metabolism of leukotriene B4 by the human neutrophil when studied in a cell superfusion model. AB - Stimulated human neutrophils are known to synthesize large quantities of 1-0 alkyl-2-acetyl-sn-glycero-3-phosphocholine (PAF) and 5,12-dihydroxy-6,14-cis-8,10 trans-transeicosatetraenoic acid (LTB4). However, in an isolated cell suspension the majority of synthesized PAF appears to remain cell associated. In addition, LTB4 is rapidly metabolized to an omega-oxidation product (20-OH-LTB4). Experiments were designed to test the hypothesis that the degree of association of PAF with the neutrophils and the metabolism of LTB4 by the neutrophils is a result of the in vitro condition used during cell activation. Here we have compared in paired experiments ionophore A23187-induced production of PAF and LTB4 by human neutrophils in a concentrated cell suspension, a diluted cell suspension and in a system in which the cells are placed on a matrix and superfused with buffer at a constant flow rate (dynamic system). There was little difference in the amount of PAF synthesized in the concentrated cell suspension and the dynamic system. However, less PAF was produced by neutrophils in the dilution system. The percent of PAF released was consistently greater in the dynamic and dilution systems than in the concentrated cell suspension. For example, more than 40% of PAF measured by incorporation of [3H]acetate or gas chromatography/mass spectrometry was released in the dynamic system and dilution systems. In contrast, less than 15% of the PAF synthesized was released from the cells in the concentrated cell suspension. 1-0-Hexadecyl-2-acetyl-3-GPC was primarily released from the neutrophils. By contrast both 1-0-hexadecyl and 1-0 octadecyl linked species of PAF were found within the cells. Exogenous PAF added to neutrophils in the dynamic or dilution systems was taken up and metabolized at a significantly lower rate than that added to cells in the concentrated cell suspension. Most of the leukotrienes synthesized by the neutrophil during A23187 stimulation were released from the cells. However, studies of LTB4 metabolism revealed differences between the dynamic and concentrated cell suspension designs. By 20 min, most of the LTB4 was recovered as 20-OH-LTB4 in the concentrated cell suspension, whereas in the dynamic system little 20-OH-LTB4 was found in the superfusate over 20 min. These experiments suggest that a large proportion of PAF synthesized by neutrophils may be released. They also suggest that the omega-hydroxylation of LTB4 by neutrophils occurs after synthesized LTB4 is released and taken back up by the cell. PMID- 2555415 TI - Platelet-activating factor and leukotriene biosynthesis in whole blood. A model for the study of transcellular arachidonate metabolism. AB - As a model to perhaps better indicate potential in vivo tissue inflammatory events, the generation of leukotriene (LT)B4, 20-OH-LTB4, sulfidopeptide LT, and platelet-activating factor (PAF) from human whole blood stimulated with zymosan was compared with that produced by isolated human neutrophils suspended either in buffer or plasma. Several reports have shown that substantial LTB4 biosynthesis could be induced after addition of zymosan to whole blood, but little was known concerning the generation of other important lipid mediators, or the cellular source of these. We have shown that, in spite of some subject variation, the zymosan-induced production of 20-OH-LTB4, LTB4, and LTE4 reached maxima within 30 to 60 min with 1.1, 2.8, and 0.60 ng/10(6) neutrophils, respectively. These concentrations would be sufficient to induce significant biologic effects. Studies with isolated cell mixtures suggested that the neutrophil was the primary source of the lipid mediators or their precursors in this system, although a number of other cell types contributed as accessory cells to the final amounts and mix of mediators produced. The ratio of neutrophils to accessory cells in mixed cell experiments dramatically modified the metabolic pattern of leukotriene generation. The concentration of LTB4 was increased in the presence of RBC and that of LTE4 when platelets were present. These results suggested that cellular cooperation and transcellular biosynthesis played a key role in the overall production of eicosanoids such as LTB4 and LTC4. The concomitant synthesis of PAF in isolated cells and in whole blood was also determined as another member of the complex lipid mediator network. Maximal production of cell-associated PAF was observed within 30 min after the initiation of phagocytosis and reached levels of 3 to 5 ng PAF/10(6) neutrophils. When other cells were present in a coincubation system, the time course for production of PAF was not altered, but maximal concentration of PAF was lower, perhaps as a result of enhanced PAF metabolism. Study of eicosanoids and other lipid mediator production in mixed cell populations provides insight into those events occurring within tissues, where cross-cell signaling and transcellular biosynthesis may occur. PMID- 2555416 TI - Identification of functional platelet-activating factor receptors in Raji lymphoblasts. AB - The binding and metabolism of platelet-activating factor (PAF) were characterized in Raji, a human Burkitt's lymphoma-derived cell line. Raji lymphoblasts readily metabolized PAF by deacetylation-reacylation at 37 degrees C, but not at 4 degrees C. Binding studies conducted at 4 degrees C demonstrated specific binding that reached saturation within 80 min. This binding was only partially reversible. Scatchard analysis of PAF binding data revealed a single class of PAF binding sites (17,800 +/- 3,600/cell) with a K of 2.3 +/- 0.3 nM. These high affinity PAF binding sites were shown to be functional receptors, as 100 pM to 1 microM PAF increased free intracellular calcium in a dose-dependent manner. The dose of PAF necessary to achieve half maximal calcium mobilization response was 6.3 nM, which was in the range of the K for the receptor calculated from the binding studies. The structurally dissimilar PAF receptor antagonists CV-3988 and BN52021 inhibited the PAF-induced calcium changes at doses that competed with PAF binding. These studies provide the first evidence for a functional PAF receptor expressed on a lymphocyte cell line. PMID- 2555417 TI - Mouse hepatitis virus 3 pathogenicity expressed by a lytic viral infection in bone marrow 14.8+ mu+ B lymphocyte subpopulations. AB - Mouse hepatitis virus type 3 (MHV3) provides an excellent model for studying viral-B lymphocyte interaction in the immune system, which plays an important role in the outcome of an acute disease. Bone marrow B lymphocyte subpopulations, at various times postinfection, were studied in genetically C57BL/6 and resistant A/J mice, infected with pathogenic L2-MHV3 and its nonpathogenic variant, YAC MHV3. B lineage cell subpopulations were identified by double immunofluorescence assays using mAb of terminal deoxynucleotidyl transferase, 14.8 and cytoplasmic (cu) or surface (su) Ig mu-chains. Results revealed diminished percentage and absolute number in the bone marrow 14.8+ mu+ B lymphocyte subpopulations, including pre-B (cu+ su-) and B (cu+ su+) cells of L2-MHV3-infected susceptible C57BL/6 mice; whereas, slight or no increase was evident in the cell subpopulations of L2-MHV3 infected resistant A/J mice or in YAC-MHV3 infected in both strains of mice. Abnormal large-sized forms of the 14.8+ mu+ cells occurred, at 48-h postinfection, in L2-MHV3-infected susceptible C57BL/6 mice only. In contrast, no change in the percentage and absolute number of precursor cells (terminal deoxynucleotidyl transferase positive) and pre pre-B cells (14.8+ mu-) were detected in all infected mice. In vitro L2-MHV3 infection of C57BL/6 bone marrow purified B lineage cell subpopulations showed that pre-B (cu+ su-) and B (cu+ su+) cells became abnormally large in size and depleted in number as a result of a productive and lytic viral replication. Low L2-MHV3 viral replication occurred in these cell subpopulations of A/J mice but no YAC-MHV3 virus was produced in the cells of both strains of mice. Pre pre-B (14.8+ mu-) cells in both strains were not permissive to L2-MHV3 or YAC-MHV3 viral replication. These results are discussed with regard to the role of humoral immunodeficiency in the pathogenic process. PMID- 2555418 TI - The productive gene for alpha-H chain disease protein MAL is highly modified by insertion-deletion processes. AB - alpha-H chain diseases (HCD) is a human lymphoproliferative disorder, characterized by the production of truncated alpha-Ig H chains, without associated L chains. In this study, we have analysed the serum protein, the alpha HCD mRNA and the rearranged alpha-HCD gene from the leukemic cells of a patient (MAL) with alpha-HCD. The abnormal MAL serum Ig consisted of short alpha 1 chains, lacking VH and CH1 domains (only CH2 and CH3 domains were present). The alpha-HCD mRNA (1.2 kb) was shorter than a normal alpha-mRNA (2 kb); the corresponding cDNA had sequences for the leader, a 84-bp sequence of unknown origin and the CH2 and CH3 exons. The establishment of the sequence of the productive alpha-HCD MAL allele revealed two major deletions; that of the VH region as well as that of the CH1 region. The JH region is altered by multiple mutations, small insertions and a duplication of the psi JH3 region. A large insert (INS1), of 360 bp (containing the 84 bp exon found in the cDNA), replaces the deleted VH region. INS1 is non-Ig related and apparently of nongenomic origin. A large second insert (509 bp), is located between the enhancer and the switch region. Insert2 contains repetitive non-Ig-related sequences and a small Ig-related sequence. All these alterations resulted in an abnormal mRNA, which comprises the leader, a 84-bp alien exon derived from INS1 and the CH2 and CH3 exons of the alpha 1-gene. PMID- 2555419 TI - Cellular and humoral immunity to varicella zoster virus glycoproteins in immune and susceptible human subjects. AB - To further delineate the immune responses that protect against serious primary varicella zoster virus (VZV) infection and inhibit viral reactivation, antibody responses and T lymphocyte reactivity to three major VZV glycoproteins, gpI, gpII, and gpIII, were studied. Individual viral glycoproteins were purified using murine monoclonal antibodies. Cellular immunity was measured by lymphocyte proliferation. Antibody responses were tested in enzyme-linked immunosorbent assays. Individual glycoproteins induced VZV-specific proliferation by mononuclear cells from 15 of 20 immune subjects. Serologic responses to the VZV glycoproteins occurred in 16 of 20 immune subjects. Of note, gpII served as a potent T and B cell antigen during both acute infection and convalescence. Cell mediated responses to the glycoprotein antigens represented proliferation by T lymphocytes and required antigen presentation by adherent mononuclear cells. These findings indicate that virally encoded glycoproteins contain epitopes that stimulate VZV-specific cellular and humoral immune responses. PMID- 2555420 TI - Congenital cytomegalovirus labyrinthitis and sensorineural hearing loss in guinea pigs. AB - Cytomegalovirus (CMV) is the leading cause of human nonhereditary congenital deafness. The pathogenesis of congenital CMV infection in the auditory system is poorly understood and no suitable animal model is currently recognized. In this study primary maternal CMV infection in guinea pigs during the first or second trimester of pregnancy resulted in congenital infection in 64% of the offspring. Of the congenitally infected neonates, 28% had significant auditory deficits. Within the inner ear, CMV infection was localized in auditory nerve spiral ganglion cells. These findings indicate that congenital CMV infection of the guinea pig results in physiologic and anatomic neuropathology similar to that seen in human infection and provide the first experimental model for congenital CMV-induced sensorineural hearing loss. PMID- 2555421 TI - A large outbreak of epidemic keratoconjunctivitis: problems in controlling nosocomial spread. AB - Between July 1985 and January 1986, 401 patients with adenovirus epidemic keratoconjunctivitis (EKC) were seen at the Illinois Eye and Ear Infirmary. Of the cases, 110 (27%) were nosocomial; the other 291 patients had community acquired infection. The highest attack rates of EKC occurred in patients attending specialty clinics; the overall attack rate among clinic patients was 4.7/1,000 clinic visits. All nosocomial cases were caused by adenovirus type 8; community acquired cases were a mixture of adenovirus types 8 and 37. Adenoviruses were isolated from conjunctival cultures up to 14 d after the onset of clinical illness. Initial efforts to prevent nosocomial transmission were unsuccessful. However, when a plan to triage all patients on entry to the infirmary and to sort patients and personnel caring for infected patients into cohorts was implemented, nosocomial transmission of EKC was promptly and effectively halted, despite the continuation of the community epidemic for another 4 mo. This outbreak clearly demonstrates the efficacy of rigorous infection control in preventing nosocomial transmission of adenovirus EKC. PMID- 2555422 TI - Investigation of an outbreak of adult diarrhea rotavirus in China. AB - In 1987 an epidemic of diarrhea associated with adult diarrhea rotavirus (ADRV) occurred in Qinhuangdao City, China, affecting more than 200 persons and causing 2 deaths. The outbreak was introduced by a person returning from an epidemic area and was spread initially to his family members and subsequently to the entire community. Adults were at greater risk of diarrhea than children 0-4 y of age and, the duration of illness increased significantly with increasing age. ADRV was identified by ELISA and electron microscopy. The electropherotypes of all positive specimens were identical, consistent with the single point-source introduction of the virus. Seroconversion was detected in 6 of 7 ill persons with a blocking ELISA. Both asymptomatic infection and person-to-person spread identified in this epidemic suggest that current emphasis on preventing waterborne transmission may not control the introduction of ADRV into new areas. The predisposition of adults for more severe disease with ADRV is similar to the pattern observed with other enteric viruses such as the Norwalk agent and hepatitis A. PMID- 2555423 TI - Effect of dose and immunization schedule on immune response of baboons to recombinant glycoprotein 120 of HIV-1. AB - To evaluate the immunogenicity of purified recombinant envelope glycoprotein of HIV-1 (rgp120) as a potential vaccine for AIDS, the antibody response of 45 baboons to rgp120 was investigated using an adjuvant (alum) and route of administration (intramuscular) suitable for humans. The primary purpose was to evaluate the effects of different doses and immunization schedules on the antibody response to rgp120 in primates. A secondary objective was to evaluate possible adverse consequences of rgp120 immunization. A liquid-phase radioimmunoprecipitation (RIP) assay for detection of rgp120-reactive antibodies revealed that rgp120 doses of 30-300 micrograms per administration resulted in nearly indistinguishable serum antibody responses. However, significant enhancement of serum antibody titers was observed when the interval between the second and third administrations was increased from 4 to 20 w. Although changing the interval significantly altered the magnitude of resulting peak titers, the kinetics of antibody formation were not changed. Thus, of the three schedules of immunization tested, none resulted in a sustained humoral immune response. The significance of the RIP assay for evaluating immune responses was confirmed by analysis showing that the percentage of immunized baboons that developed in vitro HIV-1 serum neutralizing responses was greatest in groups that also exhibited high anti-rgp120 RIP titers. Immunization with rgp120 had no significant adverse effect on any clinical or laboratory parameter monitored over the course of the study. PMID- 2555424 TI - [Effect of estrogen replacement therapy on the serum osteocalcin level in the postmenopausal and castrated women]. AB - Osteocalcin (OC), the major noncollagenous bone protein, is a vitamin K-dependent protein which is synthesized in osteoblasts. Serum OC is increased in patients with certain metabolic bone diseases, but little is known about the effects of menopause, castration, and the estrogen-replacement therapy on serum OC. In this study, we attempted to determine the serum OC level before and after menopause and castration, and the effects of estrogen on serum OC were also studied. Mean serum OC in women who underwent menopause or castration within 5 years was 4.56 +/- 1.74 ng/ml (mean +/- SD) and the concentration was significantly higher than that of premenopausal women. Serum OC was then, transiently reduced but increased again when estrogen deprived conditions lasted for more than 10 years. Estrogen replacement therapy (conjugated estrogen 0.625 mg/day for 6 months) could decrease not only serum Ca. P and ALP but also the serum OC from 4.45 +/- 1.40 to 2.97 +/- 1.43 ng/ml (p less than 0.001). There was a significant correlation between the percent changes in serum OC and ALP before and after estrogen administration (r = 0.60, p less than 0.05). PMID- 2555425 TI - [Human papilloma virus (HPV) antigens and DNA in dysplasia and carcinoma of the uterine cervix]. AB - Routinely paraffin-embedded sections of dysplasia, carcinoma in situ (CIS) and invasive (squamous) carcinoma of the cervix were studied to determine the participation of human papilloma virus (HPV) in these tissues. Morphological observation (1,059 cases) revealed condylomatous changes to reach 54% in dysplasia, 25% in CIS and 25% in invasive carcinoma. Condylomatous changes were also found to be 25 to 40% in the non-cancerous epithelia adjacent to in situ or invasive carcinomas. The immuno-peroxidase-PAP-method using anti-HPV serum was applied to 98 selected sections in which condylomatous changes were morphologically observed. HPV antigens were found to reach 56% in dysplasia, 42% in CIS and 35% in invasive carcinoma, and this result suggested that the morphologically observed condylomatous changes did not always coincide with virus maturation in the infected cells. By means of the in situ hybridization technique, HPV type-6, -11, -16 and -18 DNAs were all detected in dysplasia sections, whereas HPV type-16 DNA was demonstrated distinctively at a high rate among in situ and invasive carcinomas. PMID- 2555426 TI - [Chronological changes in sex steroids and prostaglandins receptor concentrations in rat endometrium during peri-implantation period]. AB - Successful completion of implantation requires coordinated functional synchronism of fertilized egg and endometrium. In the present study, these synchronized changes were analyzed by chronological changes in estradiol (E2), progesterone (P4) and prostaglandin E2 and F2 alpha (PGE2, PGF2 alpha) receptor (R) concentrations in various sites in rat endometrium during the peri-implantation period. Unilaterally pregnant horn prepared by tubal ligation was also used. The results obtained: Nuclear E2R (E2RN) increased in the first 5 days of the fertilized cycle but decreased after that in the pregnant horn. P4RN, on the other hand, continued to increase throughout the period studied. PGE2-R remained low until the 6th day but thereafter showed a rapid increase. PGF2 alpha-R was significantly higher in the pregnant horn but showed a fall after day 6. Cyclooxygenase distribution, studied by 14C-indomethacin binding, showed an increase in the peri-implantation period. The synchronous changes required for successful implantation, therefore, involve complex changes in steroids as well as the prostaglandin receptor concentration in the endometrium around the implantation tissue. PMID- 2555427 TI - Interrelation among endogenous catecholamines, prostaglandin F2 alpha and prolactin in last trimester and during parturition. AB - To investigate the interrelation among endogenous catecholamines, prolactin and prostaglandin F2 alpha, we measured the plasma levels of dihydroxyphenylalanine (DOPA), norepinephrine (NE), catecholamine metabolites (3,4-dihydroxyphenylacetic acid [DOPAC], 3,4-dihydroxyphenylethyleneglycol [DOPEG], cyclic 3',5'-adenosine monophosphate (c-AMP), prostaglandin F2 alpha (PGF2 alpha) and prolactin (PRL) in 119 patients, before, during and after a normal labor and delivery, and in the early puerperium. In late pregnancy, DOPAC concentrations were significantly increased, and reached peak values before the onset of labor in the group who went into spontaneous labor. The DOPA, NE and DOPEG levels remained unchanged before labor. The plasma concentrations of NE, c-AMP and PGF2 alpha were significantly increased during labor. There was a positive correlation between plasma NE and c-AMP (r = 0.584, p less than 0.001), and between NE and PGF2 alpha (r = 0.401, p less than 0.001) during labor. In contrast, plasma PRL significantly decreased during labor and immediately after delivery. There was a statistically significant inverse relationship between PRL and NE (r = -0.392, p less than 0.001) and between PRL and PGF2 alpha (r = -0.523, p less than 0.001) during labor, but there was no statistical correlation among PRL, DOPA and DOPAC. In conclusion, these data suggest that the dopaminergic activity in the maternal circulation increases before the onset of labor, and the PRL release from the maternal pituitary gland during labor is probably not controlled primarily by dopaminergic neurons and may be suppressed by other mechanisms, such as the stress of labor and/or the influence of PGF2 alpha. PMID- 2555428 TI - A rational classification of angiofibromas of the post nasal space. AB - Angiofibromas of the post nasal space have the potential to extend in all directions along predetermined pathways with a definite mode and chronological pattern. In order to understand their clinical behaviour, methods of treatment and prognosis, a system of clinical staging based on conventional and tomographic observations has been found useful in this series of cases. The classification has been evolved according to the extent of the tumour involvement but also the age of onset, duration, size and area of encroachment including the surgical approach and consequent prognosis. Deficiencies in the earlier classifications are discussed. PMID- 2555429 TI - Granular cell tumour of the larynx in childhood. AB - The youngest patient reported to date with granular cell tumour of the larynx is presented, with a review of other reported cases in childhood. The basis of histological diagnosis of granular cell tumour is described. PMID- 2555430 TI - Recovery frequencies of herpes simplex virus types 1 and 2 from symptomatic and asymptomatic genital herpes cases and antiviral sensitivities of isolates. AB - Herpes simplex viruses were isolated from 40.8 to 56.0 per cent of the patients with genital herpes. The frequency of recovery seemed to be higher in females than in males, particularly during the first episode of infection. Asymptomatic shedding of the virus from female genitalias was approximately 0.7 per cent. Herpes simplex virus type 2 represented 98.4 per cent of all isolates and the remaining isolates were type 1. These isolates exhibited a wide range of sensitiveness to 9-(2-hydroxyethoxymethyl) guanine (acyclovir) as demonstrated by antiviral inhibition assay and no single strain exhibited high in vitro drug ID50 value. PMID- 2555431 TI - In individual T cells one productive alpha rearrangement does not appear to block rearrangement at the second allele. AB - The T cell lymphoma line BW5147 has rearranged TCR alpha chain genes segments on both the homologous chromosomes: one is functional (V alpha 1) and the second (V alpha 16.1) is a pseudogene. The extreme 3' position of the V alpha 16.1 gene segment in the V alpha locus allows us to recognize rearrangements of most V alpha gene segments using the V alpha 16 probe as a marker. The absence of the genomic V alpha 16.1 gene fragment in mature thymocytes, antigen-specific T cells, and in more than two-thirds of the peripheral T cells suggests that most T lymphocytes rearrange both alpha loci. It appears that productive alpha chain rearrangement on one allele probably does not block a subsequent rearrangement on the other alpha locus. PMID- 2555432 TI - Molecular pathogenesis of neural lesions induced by poliovirus type 1. AB - Using in situ hybridization techniques for viral RNA and employing a specific riboprobe, we have detected virus in neural cells of monkeys infected with poliovirus type 1 (PV-1) by the intraspinal route. In monkeys paralysed after inoculation of a neurovirulent revertant of PV-1/Sabin strain, viral RNA was detected in motor neurons and their processes, and in polymorphonuclear and small neural cells. Quantitative in situ hybridization provided evidence of viral replication in individual cells suggesting that the death of motor neurons was due to the direct effect of poliovirus replication in these cells. The histological study of neural lesions of monkeys paralysed after infection with the attenuated Sabin strain of PV-1 revealed two major differences compared to monkeys infected with a virulent strain: (i) the number of destroyed motor neurons was reduced and limited to the site of inoculation and (ii) the inflammatory reaction was localized but more intense. An account is given of the difference in histopathology induced by virulent and attenuated strains of PV-1 in the central nervous system. PMID- 2555433 TI - Hydrolysis of a series of synthetic peptide substrates by the human rhinovirus 14 3C proteinase, cloned and expressed in Escherichia coli. AB - The 3C proteins of several picornaviruses, including poliovirus, foot-and-mouth disease virus (FMDV) and encephalomyocarditis virus (EMCV), have been demonstrated to be cysteine-type proteinases, involved in the processing of the respective polyproteins expressed by the monocistronic RNA genome. Nucleotide sequencing data have indicated that the human rhinovirus 14 (HRV-14) RNA genome encodes a homologous 3C protein. The HRV-14 3C protein was purified to homogeneity from Escherichia coli expressing the cloned 3C genomic fragment. The enzyme was assayed against peptides corresponding to those residues, predicted (by nucleotide sequencing data) to occur at authentic cleavage sites within the polyprotein. The peptides representing the 1B/1C, 2A/2B, 2C/3A, 3A/3B, 3B/3C and 3C/3D cleavage sites, where proteolysis was predicted to occur at a Gln-Gly junction, were all cleaved by the 3C proteinase. The hydrolysis was shown (by reverse phase fast protein liquid chromatography and amino acid analysis) to occur specifically at the Gln-Gly bond in each of the peptides. The ready availability of such convenient substrates facilitated the further characterization of the 3C proteinase. By contrast, peptides corresponding to the predicted 2B/2C and 1C/1D cleavage sites, where the processing was presumed to occur at a Gln-Ala or Glu-Gly bond respectively, were not cleaved by the 3C proteinase. The ability of the HRV-14 3C proteinase to hydrolyse the synthetic peptides was inhibited if a Cys----Ser(146) mutation was introduced into the protein. Studies with known proteinase inhibitors substantiated the conclusion that the HRV-14 3C protein appears to be a cysteine proteinase and that the Cys residue at position 146 may be the active site nucleophile. The HRV-14 3C proteinase probably plays an important role, analogous to that implied for the poliovirus 3C proteinase, in the replication of the virus and thus represents a potential target for antiviral chemotherapy. PMID- 2555434 TI - A comparative analysis of the sequence of the thymidine kinase gene of a gammaherpesvirus, herpesvirus saimiri. AB - We present the nucleotide sequence of a region from the genome of the A + T-rich gammaherpesvirus, herpesvirus saimiri (HVS), which includes the coding sequences for the viral thymidine kinase (TK) gene. The organization of genes in this region resembles the homologous region of the Epstein-Barr virus (EBV) genome and is very compact, using overlapping coding sequences and with nucleotides shared by initiation and termination codons of neighbouring reading frames. The HVS TK is predicted to contain a 527 residue polypeptide with the first part of the presumptive nucleotide-binding site [(L, I, V)(F, Y)(I, L)(D, E)(G)(X)(X)(G)(L, I, V, M)(G)(K)(T, S)(T, S)] located at residues 212 to 224. This motif is close to the amino terminus of the TK polypeptides of alphaherpesviruses and the polypeptides of the cellular and poxvirus-encoded enzymes. The corresponding reading frame of the human gammaherpesvirus (EBV) also has a long amino-terminal extension but significant amino acid sequence similarities between the HVS and EBV sequences are not observed until the region of the nucleotide-binding site. Comparisons of these homologous carboxy-terminal sequences of the HVS- and EBV encoded proteins with those from six alphaherpes viruses and proteins encoded by Marek's disease virus (MDV) and the herpesvirus of turkeys (HVT) confirm that the HVS and EBV sequences are products of a distinct lineage. The sequences of alphaherpesvirus enzymes than to those of HVS and EBV. Comparison of these 10 highly divergent TK sequences extends and refines the identification of essential features of this family of herpesvirus enzymes and defines 19 positions at which all sequences have identical residues. PMID- 2555435 TI - Nucleotide and predicted amino acid sequences of the Marek's disease virus and turkey herpesvirus thymidine kinase genes; comparison with thymidine kinase genes of other herpesviruses. AB - In this paper we present the nucleotide sequences of the thymidine kinase (TK) genes of two avian herpesviruses: a highly oncogenic strain of Marek's disease virus (MDV strain RB1B) and its serologically related vaccine virus, the herpesvirus of turkeys (HVT strain Fc-126). The predicted coding regions of the two genes are 1029 and 1050 nucleotides respectively, corresponding to polypeptides of 343 and 350 amino acids in length. Putative nucleotide- and nucleoside-binding sites have been identified within the two predicted amino acid sequences. The MDV and HVT TK amino acid sequences exhibit 58.2% amino acid identity. Comparison with other available herpesvirus TK sequences reveals a greater homology to those of the alphaherpesviruses than to those of the gammaherpesviruses. No overall homology was found when compared with the chicken cytoplasmic TK sequence. PMID- 2555436 TI - Location and characterization of the bovine herpesvirus type 2 thymidine kinase gene. AB - The precise genomic location and the nucleotide sequence of the bovine herpesvirus type 2 (bovine herpes mammillitis virus) thymidine kinase (TK) gene have been determined. The genomic location of the TK gene was found to be in a similar position to that of herpes simplex virus. The coding region consists of 918 bases, which is slightly smaller in length than other reported herpesvirus TK genes. However with an Mr of 38,108 the individual protein is similar in size to other herpesvirus TK enzymes. Despite there being only limited overall sequence homology with the TK genes of other herpesviruses, there are several regions of extensive homology at the amino acid level. PMID- 2555437 TI - The herpes simplex virus type 2 (HG52) variant JH2604 has a 1488 bp deletion which eliminates neurovirulence in mice. AB - The herpes simplex virus type 2 (HG52) deletion variant JH2604 is avirulent (LD50 greater than 10(7) p.f.u./mouse) for mice compared to the parental wild-type virus (LD50 less than 10(2) p.f.u./mouse) and fails to replicate in vivo. JH2604 has a 1488 bp deletion within the 3 kb BamHI v fragment (0 to 0.02 and 0.81 to 0.83 map units) which removes one copy of the 17 bp direct repeat DR1 element of the 'a' sequence and terminates 522 bp upstream of the 5' end of the immediate early gene 1. In vivo selection after transfection of intact JH2604 DNA with the BamHI g (v + u) joint fragment of HG52 results in the isolation of wild-type virus with an LD50 of less than 10(2) p.f.u./mouse. These results show that a 1488 bp sequence within the terminal portion of the genome long repeat region confers neurovirulence on strain HG52. PMID- 2555438 TI - The terminal protein gene 2 of Epstein-Barr virus is transcribed from a bidirectional latent promoter region. AB - The intact terminal protein genes (TP1 and TP2) of Epstein-Barr virus (EBV) are created upon infection by circularization of the linear viral genome at its terminal repeats. The structure of the 1.7 kb TP2 latent mRNA has been determined by cDNA analysis and Northern blotting, revealing its close relation to TP1 mRNA. The 1.7 kb transcript is expressed from a different promoter and has a different 5' exon from TP1 but is also spliced across the terminal repeats. The last eight exons are common to the TP1 and TP2 RNAs. The TP2 promoter is 3.3 kb downstream of the TP1 promoter and is part of a bidirectional latent EBV promoter region transcribing the TP2 and the latent membrane protein RNAs in opposite directions. PMID- 2555439 TI - Identification of 21 genes of infectious laryngotracheitis virus using random sequencing of genomic DNA. AB - DNA from infectious laryngotracheitis virus (ILTV) was randomly sheared and cloned into the M13 bacteriophage. Clones containing ILTV DNA were sequenced and the predicted amino acid sequences were compared to the known sequences of other herpesviruses using computer analysis. Twenty-one ILTV genes were identified, 20 by comparison to varicella-zoster virus and 19 by comparison to herpes simplex virus type 1; only 12 genes, giving consistently lower homology scores, were found by comparison with the gammaherpesvirus Epstein-Barr virus, indicating that ILTV sequences bear greater similarity to other alpha- than to gammaherpesvirus sequences. PMID- 2555440 TI - Human papillomavirus type 56: a new virus detected in cervical cancers. AB - A new human papillomavirus type (HPV-56) was identified by low stringency Southern blot analysis with an HPV-31 DNA probe, in a cervical intraepithelial neoplasm (CIN). The DNA of this virus was molecularly cloned and shown to be a new HPV type based on the absence of cross-reactivity to HPV types 1 to 55 under high-stringency hybridization conditions. At low stringency, HPV-56 was most related to HPV types 30 and 45. The deduced organization of the open reading frames of HPV-56, from hybridization and partial nucleotide sequence analyses, reveals a typical HPV genome. HPV-56 was detected in two of 464 normal cervical tissues, in five of 227 cervical condylomas and CIN, and in two of 84 invasive cancers of the cervix. PMID- 2555441 TI - Typing of human rhinoviruses based on sequence variations in the 5' non-coding region. AB - Unambiguous assignment of restriction enzyme patterns to six individual serotypes of human rhinovirus was accomplished after amplification of a 380 bp DNA fragment derived from the 5' non-coding region. This was possible even though serotypes 1A and 1B and serotypes 2 and 49 differed only at 10 and 15 positions respectively. The method utilizes the conserved and variable components of this part of the genome and provides the basis for a simple and rapid method for typing of human rhinoviruses. PMID- 2555442 TI - Use of anticontamination primers in the polymerase chain reaction for the detection of human papilloma virus genotypes in cervical scrapes and biopsies. AB - A reliable application of the polymerase chain reaction (PCR) for detection of the human papilloma virus (HPV) genotypes in cervical smears and biopsies was developed. Primers flanking the HPV cloning site were used to avoid detection of cloned HPV plasmids. These anticontamination primers were used for the specific detection of HPV 6, 11, 16, 18, and 33 in cervical scrapes that had been tested previously for HPV with a combined modified filter in situ hybridization (modified FISH) and dot blotting procedure. The PCR appeared to be superior. Two groups of women were screened for HPV genotypes. Group A consisted of women belonging to a regularly screened population, and group B contained women attending a gynaecological clinic. It appeared that the overall prevalence of HPV in cytologically normal scrapes in the first group was 6%, whereas in the second group 12% was found. In scrapes with cytological dysplasia, the prevalence of HPV in group A and B was approximately 40% and 60%, respectively. HPV 16 was present predominantly. In biopsies of squamous cell carcinomas of the cervix uteri, an HPV prevalence rate of 90% was found, all of which contained only HPV 16 and 18. These data indicate an important role for HPV detection in the screening of cervical scrapes to identify women with an increased risk of cervical cancer. PMID- 2555443 TI - Adenovirus type 7 infections in children in New South Wales, Australia. AB - This study describes our experience of adenovirus type 7 infection in children in New South Wales, Australia. Some aspects of the epidemiology of the infection are also recorded. A community outbreak, a hospital-ward outbreak, sporadic cases, and data from a centralised registry are described. Results of genome-type identification using restriction endonucleases are given. Adenovirus type 7b infection was associated with significant mortality and points to the need, previously expressed, for an adequate vaccine for high-risk infants. Continued surveillance of adenovirus type 7 infections worldwide is necessary to identify genome types so that appropriate vaccines can be developed. PMID- 2555444 TI - DNA dot-blot hybridization implicates human papillomavirus type 11-DNA in epithelioma cuniculatum. AB - Epithelioma cuniculatum is a rare, low-grade squamous cell carcinoma, belonging to the group of verrucous carcinomas. Evidence is presented that suggests that human papilloma virus type-11 may be involved in the pathogenesis of epithelioma cuniculatum. HPV-DNA was detected by dot-blot hybridization with 32P-labelled probe DNA. Plasmids containing HPV-DNA were isolated by the benzoylated naphthoylated DEAE cellulose method (BND-method), an improved procedure for routine detection of HPV-DNA. PMID- 2555445 TI - Predilection of a nasopharyngeal carcinoma-derived isolate of Epstein-Barr virus for infection of specific subsets of B lymphocytes. AB - It is important to know whether there are variants of Epstein-Barr virus (EBV) with biological properties that are different from the prototype viruses that have been studied in detail, such as P3HR-1 and B95-8. We have studied an EBV isolate derived from a nasopharyngeal carcinoma (NPC) tumor, designated NPC-EBV. We have examined the target B lymphocytes infected and growth-transformed with NPC-EBV as compared with two common EBV isolates, B95-8 and AG876 EBV, for stage of maturation using antibodies to several immunoglobulin chains. Typing of the NPC-EBV transformed lymphoblastoid cell lines revealed the predilection of the NPC-EBV isolate to infect immature B lymphocytes. This was not the case for the B95-8 and AG876 isolates. The reason for the predilection of NPC-EBV for immature B lymphocytes remains to be explored further. However, these results may be important in understanding the pathophysiology of EBV-associated diseases. PMID- 2555446 TI - Tropism and histopathology of the D, B, K, and MM variants of encephalomyocarditis virus. AB - Variants of encephalomyocarditis virus (EMCV), which are immunologically indistinguishable by hyperimmune serum, produce different disease syndromes in mice. For instance, in ICR Swiss male mice, EMCV-B produces no overt illness, EMCV-MM produces severe neurological signs followed by death, EMCV-D destroys pancreatic beta cells producing a disease syndrome resembling insulin-dependent diabetes mellitus, and EMCV-K is lethal but produces no overt signs of infection. The present study was done to determine the tissue tropism and histopathology of each of these EMCV variants in the ICR Swiss mouse model. The data show the highest concentrations in the following organs: EMCV-D in the pancreas, EMCV-B in the pancreas, EMCV-MM in the cerebrum, and EMCV-K in the medulla/brainstem. They also show that the pathological lesions produced by each variant correlate well with viral titers. PMID- 2555447 TI - Combined effects of acyclovir and human interferon-alpha on herpes simplex virus replication in cultured neural cells. AB - The combined effects of Acyclovir [9-(2'-hydroxyethoxymethyl)guanine; ACV] and human interferon-alpha (IFN-alpha) on replication of the herpes simplex virus type I (HSV-1) were determined in human neural cell lines, neuroblastoma (IMR), glioblastoma (118MGC), and glioma (U251MG). HSV-1 grew well in all these cells, with final yields of more than 1 x 10(6) PFU/ml. In terms of virus-yield reduction, ACV was found to be highly effective in IMR, moderately effective in U251MG, but ineffective in 118MGC. By contrast, IFN-alpha reduced the virus yield significantly in 118MGC and in U251MG, but did not in IMR. Combined application of ACV and IFN-alpha strongly inhibited the virus replication in all three cell lines with various degrees of synergism or additive effect. These results were also confirmed by immunofluorescent examinations. The sensitivity of HSV-1 to ACV or IFN-alpha was found to be different among the three different cell types. By combining the two agents, the virus growth was strongly suppressed in all the cells. These results suggest the importance of combination therapy for severe type of herpes simplex encephalitis in clinical practice. PMID- 2555448 TI - Validation of a first-generation Epstein-Barr virus vaccine preparation suitable for human use. AB - The efficacy of a new vaccine preparation against Epstein-Barr (EB) virus was investigated in cotton-top tamarins. The vaccine consists of fast protein liquid chromatography-purified EB virus membrane antigen glycoprotein of 340 Kd (MA gp340) mixed with a synthetic muramyl dipeptide adjuvant emulsified in squalane containing a pluronic polymer; it is suitable for both scaled-up batch production and eventual administration to man. Vaccinated tamarins rapidly developed ELISA detectable high titre antibodies to MA gp340, and their sera became strongly EB virus-neutralising. After challenge with a massive 100% carcinogenic dose of EB virus, the vaccinated tamarins had a strikingly low level of circulating EB virus carrying mononuclear cells, in contrast to a control animal, and remained entirely free of tumours. This first-generation vaccine has thus been validated in experimental animals and the way opened for a phase I human trial. PMID- 2555449 TI - A molecular modelling study of the interaction of noradrenaline with the beta 2 adrenergic receptor. AB - A model of the beta 2-adrenergic receptor binding site is built from the primary structure of the receptor, experimental evidence for key binding residues and analogy with a homologous protein of partially determined structure. It is suggested that residues Trp-109, Thr-110 and Asp-113 are involved in ligand binding. Noradrenaline is successfully docked into this model, and the results of an INDO molecular orbital calculation on the complex indicate that a charge transfer interaction between Trp-109 and noradrenaline is possible. PMID- 2555450 TI - Elevated plasma concentrations of C-flanking gastrin-releasing peptide in small cell lung cancer. AB - Many small-cell lung cancers (SCLCs) produce gastrin-releasing peptides (GRPs) (mammalian bombesin) but the plasma concentration of GRP is rarely elevated, possibly because of its rapid elimination. We developed a radioimmunoassay for the C-terminal flanking peptide of proGRP and measured its concentration in plasma from 71 patients with SCLC, in 27 healthy subjects and in 49 patients with other diseases including lung carcinomas. In addition, we studied the molecular size of immunoreactive C-flanking peptide in two SCLC cell lines and in plasma from SCLC patients. The concentration of immunoreactive C-flanking peptide in normal subjects and in control patients did not exceed 10 pmol/L and 26 pmol/L, whereas 72% of the SCLC patients had C-flanking peptide concentrations above 10 pmol/L. In patients with extensive disease (n = 35) the median concentration was 71 pmol/L (range, 10 to 940). ProGRP C-flanking peptide levels paralleled the clinical course in 12 patients. The molecule(s) responsible for the immunoreactivity had a molecular size of about 8 to 10 kd in both patient plasma and tumor cell lines, suggesting that the measured peptide(s) represented major fragment(s) if not the entire C-flanking peptide of proGRP. Thus this peptide(s) seems to be a useful marker for SCLC. PMID- 2555451 TI - A phase II clinical trial of adoptive immunotherapy for advanced renal cell carcinoma using mitogen-activated autologous leukocytes and continuous infusion interleukin-2. AB - Forty patients with metastatic, recurrent, or unresectable renal cell carcinoma were entered into a study of the therapeutic efficacy of adoptive immunotherapy using periodate (IO4-) and interleukin-2 (IL2)-activated autologous leukocytes and continuous infusion low-dose IL2. Patient survival was also examined. The first 15 consecutive patients were enrolled in protocol A without an IL2 priming phase and the following 25 patients were entered in protocol B where a 5-day priming phase was initiated before leukapheresis. A maintenance regimen consisted of either 3 x 10(6) units of recombinant interferon-alpha (rIFN-alpha), three times per week only or together with leukapheresis and infusion of IO4-/IL2 activated cells and 2 days of continuous infusion IL2 per month. Thirty-four patients completed the protocol treatment. Four patients were removed from the study owing to rapid tumor progression and two patients died while receiving treatment. The clinical response rate was 22%: two patients had a complete response and five patients had a partial response. Among the 25 patients who had no clinical response, 11 patients had either a mixed response or stabilization. Neither response, response duration, nor site response correlated with the total dose of IL2 administered or the number of activated killer cells infused. Patients who received maintenance therapy had longer survival times than patients who did not receive such therapy. All toxicity and side effects associated with IL2 treatment were transient and resolved after discontinuation of the drug. Patients on maintenance therapy tolerated both rIFN-alpha and monthly infusions of activated killer cells and IL2 well. This study confirms the concept of adoptive immunotherapy as a new treatment approach for advanced renal cell carcinoma and suggests that maintenance therapy may prolong survival time. PMID- 2555452 TI - A novel pharmacodynamically based approach to dose optimization of carboplatin when used in combination with etoposide. AB - Thrombocytopenia, the dose-limiting toxicity of carboplatin, is manageable and predictable with the dosing equation: Dose (mg/m2) = (0.091) (creatinine clearance)/(body surface area) (desired percentage change in platelet count) + 86. We used this equation to dose patients receiving carboplatin (day 1) and etoposide (80 mg/m2 days 1 to 3). An initial cohort of 14 patients with non-small cell lung cancer (NSCLC) were treated with 75% of the calculated dose of carboplatin (29 evaluable courses) as a precaution against added myelosuppression from combination chemotherapy. The observed reduction in platelets was essentially equal to the reduction in platelets predicted if patients had received carboplatin alone. Subsequently, a second cohort of 20 evaluable patients with NSCLC received the full calculated dose of carboplatin and etoposide (51 evaluable courses). There was a linear relationship between observed (o) and predicted (p) reductions in platelets. With full-dose carboplatin in combination with etoposide, there was a significantly greater carboplatin dosage administered, greater reduction in platelets, and lower platelet nadir. Among 34 evaluable patients (80 courses) there was one complete response (CR) and four partial responses (PRs) for an overall response rate of 15% (90% confidence +/- 9%). The median duration of survival for responders was 336+ days and for nonresponders was 204+ days. Therapy was well tolerated. This study, in addition, supports our concept of individualized dosing of carboplatin and the underlying pharmacokinetic/pharmacodynamic relationships and represents an interesting pharmacodynamic and quantitative approach to studying potential drug-drug interactions and defining appropriate dosages for combination chemotherapy. PMID- 2555453 TI - The modulation of astrocytic differentiation in cells derived from a medulloblastoma surgical specimen. AB - Medulloblastomas are cerebellar tumors which are primarily composed of sheets of uniform, small malignant cells and may have astrocytic, neuronal or no features typical of these cell types. The assessment of astrocytic differentiation in medulloblastoma rests largely on the detection in malignant cells of glial fibrillary acidic protein (GFAP), a marker present in the later stages of normal astrocyte differentiation. It is still not known whether cells that do not contain GFAP in medulloblastomas with astrocytic differentiation correspond to highly proliferative astrocyte progenitors in maturation arrest at earlier stages of differentiation. The purpose of the current study was to examine whether cells in short term culture derived from a medulloblastoma tumor specimen with astrocytic differentiation were of the astrocytic lineage and if so, whether they represented proliferative astrocyte progenitors which would morphologically and antigenically mature in response to differentiating agents. A portion of tumor specimen from a 10-month-old child with recurrent posterior fossa medulloblastoma (RB2) that contained GFAP focally in tumor cells was grown in monolayer culture. We examined cellular structure and appearance of western immunoblotting and immunohistochemical studies for GFAP and neuron-specific enolase (NSE) in RB2 cells before and after treatment with retinoic acid (RA) and dibutyryl cyclic AMP (dBcAMP). RB2 in culture consisted of small polygonal cells (93%), large flat cells (3%), and polygonal cells with cytoplasmic processes (4%). In untreated RB2, 30% of cells expressed GFAP and staining for NSE was negative. RA treatment produced flattened cells and decreased GFAP. DBcAMP reversibly induced fine cytoplasmic processes containing GFAP in 85% of cells within 96 h. Neither agent induced NSE. The results suggest that cultured cells which are derived from a medulloblastoma with astrocytic differentiation do not spontaneously differentiate but that treatment with dBcAMP suppresses proliferation, enhances cytoplasmic process formation and increases cytoplasmic GFAP. Cells in culture and in medulloblastoma tumor specimens which do not contain GFAP may represent astrocyte progenitors in maturation arrest. PMID- 2555455 TI - Combination chemotherapy with cyclophosphamide, doxorubicin and vincristine in the treatment of stage III-IV small-cell lung cancer. AB - Twenty patients with histologically confirmed small-cell lung cancer were treated with cyclophosphamide 1000 mg/m2 i.v. on day 1, vincristine 1.4 mg/m2 i.v. day 1, and adriamycin 50 mg/m2 i.v. on day 1. This protocol was repeated every 21 days. Out of 17 evaluable patients 2 obtained a complete response (12%) with a mean duration of 11 months, 4 patients achieved a partial response with a mean duration of 6.3+ months, and 1 had a minimal response of 7.2 months. Two patients had a stabilization which lasted a mean of 4+ months, while 8 patients progressed. Although the mean survival was higher in responders than in non responders, the difference in survival time was not statistically significant. The treatment was quite well tolerated with hematological toxicity in 78% of cases, oral and gastrointestinal toxicity in 83%, alopecia in 78%, and neurotoxicity in 16% of cases. PMID- 2555454 TI - Glioblastoma multiforme of the cerebellum five decades after irradiation of a cerebellar tumor. AB - A 70-year-old woman is reported who had glioblastoma multiforme of the cerebellum 52 years after radiation therapy to a midline cerebellar tumor. Seven similar reported cases are reviewed. Dedifferentiation of astrocytoma to glioblastoma and the role of radiation therapy are discussed. PMID- 2555456 TI - Monosynaptic excitatory amino acid transmission from the posterior rhombencephalic reticular nucleus to spinal neurons involved in the control of locomotion in lamprey. AB - 1. The reticulospinal neurons in the lamprey posterior rhombencephalic reticular nucleus (PRRN) and their projections to different types of spinal neurons have been investigated by the use of simultaneous paired intracellular recordings from one pre- and one postsynaptic cell. PRRN is of particular importance for the initiation of locomotion. 2. Intracellular stimulation of single PRRN neurons produced monosynaptic excitatory postsynaptic potentials (EPSPs) in simultaneously recorded motoneurons and spinal premotor interneurons of both the excitatory and inhibitory type. Individual PRRN neurons produced EPSPs in several different types of target cells, as revealed by signal averaging. Each single PRRN neuron had extensive monosynaptic connections to approximately 73% of the motoneuronal population. Conversely, several PRRN neurons converge on individual spinal neurons. The average amplitude of the EPSPs was 0.43 +/- 0.40 (SD) mV. The EPSPs varied in time course (time to peak = 7.5 +/- 2.8 ms; duration at one-half peak amplitude = 21.9 +/- 18.1 ms). 3. The EPSPs produced by reticulospinal cells were composed of either exclusively chemical, exclusively electrical, or mixed chemical and electrical components. The electrical EPSPs remained when the ordinary physiological solution was substituted for one without Ca2+ but with Mn2+. The chemical component of the EPSPs was always depressed when a broad spectrum excitatory amino acid (EAA) antagonist, such as kynurenic acid, was applied, suggesting that the chemical component was because of EAA transmission. The chemical EPSP could have two components, one late, suppressed by N-methyl-D aspartate (NMDA) antagonists, and one early because of activation of kainate/quisqualate receptors. 4. Three-dimensional reconstructions of Lucifer yellow-filled PRRN neurons were performed with a confocal laser scanning microscope. PRRN neurons producing monosynaptic excitatory amino acid EPSPs were found to have a fusiform cell body located near the surface of the fourth ventricle and an extensive fanlike dendritic tree extending to the ventral and lateral margin of the brain stem within the basal plate. The axons descend in the lateral funiculi of the spinal cord. 5. PRRN neurons utilizing EAA transmission are active during fictive locomotion. They presumably initiate and reinforce ongoing spinal locomotor activity by monosynaptically increasing the general excitability of the spinal premotor interneurons of the spinal locomotor networks by means of their extensive divergent and convergent monosynaptic connections. PMID- 2555457 TI - Neuromuscular transmission in diabetes: response to high-frequency activation. AB - In searching for the cellular correlates of diabetic neuropathy, we examined the response to tetanic stimulation of diabetic neuromuscular junctions and of age matched controls. The experiments were performed on the soleus nerve muscle preparation of the rat in which diabetes was induced by streptozotocin. Tetanic potentiation was substantially lower in diabetic rats. In addition, it was found that the diabetic neuromuscular junction is more resistant to high-frequency stimulation than normal age-matched controls, in which such stimulation causes a progressively increasing number of failures in synaptic transmission. Tetanic failures cannot be predicted from the stochastic properties of transmitter release and are due to propagation block of action potentials into the nerve terminals. The resistance of diabetic nerves to tetanic stimulation is a function of the duration of diabetes; the earliest significant difference between the number of tetanic failures in diabetic and normal age-matched controls was observed after 19 d of diabetes, and this difference grew with increased duration of diabetes. The resistance to tetanic stimulation in diabetic rats is reversed by insulin in vivo but not in vitro. Elevation in extracellular [K+] increases the number of tetanic failures in both diabetic and normal preparations. Furthermore, elevating extracellular [K+] to 8.5 mM brings the number of tetanic failures into the range of tetanic failures in normal nerves. This finding is consistent with the hypothesis that differences in extracellular [K+] accumulation during high-frequency stimulation are responsible for the diabetic nerve's relative resistance to high-frequency stimulation. The lower number of failures corrects only partially the impaired neuromuscular transmission in the diabetic state, and there is an overall reduction in tetanic potentiation in diabetes. PMID- 2555458 TI - Brain cytochrome oxidase: purification, antibody production, and immunohistochemical/histochemical correlations in the CNS. AB - Cytochrome oxidase (CO) is a mitochondrial energy-generating enzyme used in brain studies as a marker of neural functional activity. The activity of CO in different brain regions, revealed histochemically, is distributed nonhomogeneously but in distinct patterns. Localized differences in CO activity could arise from localized differences in enzyme amount or from localized regulation of enzyme turnover number (molecular activity). To distinguish between these alternatives, we used antibodies against purified calf brain CO to assess the immunohistochemical distribution of CO amount (protein immunoreactivity) in several brain regions. Calf brain mitochondria (synaptic and nonsynaptic populations) were isolated from gray matter homogenates by differential centrifugation. CO was purified from detergent extracts of the mitochondria by cytochrome c-Sepharose 4B affinity chromatography. Antisera against the purified CO were raised in rabbits. The antibodies reacted specifically with CO, predominantly subunit IV, in SDS immunoblots. The antibodies did not react in SDS immunoblots with any other proteins solubilized from mitochondria or caudate nucleus but did cross-react with brain CO from other mammalian species and with bovine heart CO. The immunohistochemical distribution of CO amount matched the histochemical distribution of CO activity in all regions tested, including the monkey hippocampus and the mouse olfactory bulb, somatosensory (barrel) cortex, and cerebellum. Thus, the amount of CO in neural tissue is distributed in the same nonhomogeneous pattern as the histochemical activity of CO. The results suggest that mechanisms exist by which CO molecules are selectively distributed within neurons to meet local metabolic demands posed by neural functional activity. PMID- 2555459 TI - Differential effects of nerve transection on the ACh and GABA receptors of chick ciliary ganglion neurons. AB - Chick ciliary ganglion neurons have nicotinic acetylcholine receptors (AChRs) that mediate chemical transmission through the ganglion, and GABAA receptors of unknown significance. Previous experiments examining the role of cell-cell interactions in regulating neuronal AChRs have shown that postganglionic axotomy of ciliary ganglia in newly hatched chicks causes a 10-fold decline in total AChRs within 5 d compared with unoperated contralateral ganglia and that preganglionic denervation causes a 3-fold decline within 10 d. Many of the AChRs are known to be intracellular; of those present on the cell surface, only a small fraction appears to be functionally available normally. In the present experiments, the effects of the operations on functional AChRs and GABAA receptors in the plasma membrane of the neurons were examined by removing the ganglia 5 d after axotomy or 10 d after denervation, dissociating them into single cells, and immediately measuring their ACh and GABA sensitivities with intracellular recording techniques. The ACh sensitivity of axotomized ciliary ganglion neurons was reduced 10-fold compared with neurons from unoperated contralateral ganglia of the same chicks. The reduction could be largely accounted for by a decrease in the maximum response and did not arise from a change either in the dose-response curve or the acetylcholinesterase activity of the neurons. Autoradiographic studies using a radiolabeled anti-AChR monoclonal antibody also demonstrated a substantial decrease in the total number of surface AChRs associated with axotomized neurons. In contrast, axotomy had no unilateral effect on the GABA response.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555460 TI - Regulation of tension on hair-cell transduction channels: displacement and calcium dependence. AB - An epithelial preparation of the bullfrog sacculus was used to characterize the initial rate of the adaptation mechanism in hair cells and its dependence on displacement and calcium. The I(X) curve relating transduction current and bundle displacement shifted along the X-axis without substantial change in slope, as previously observed, suggesting that adaptation involves a change in the attachment point of the elastic element connected to ion channels. If the "tip links" model of transduction is correct, this implies that one end of the link moves along the side of the stereocilium. The rates were highly asymmetric: in the tensioning direction the rate was roughly constant at 1-2 microns/sec (calculated as motion along a stereocilium); this is similar to that of myosin on actin. In the relaxing direction it appeared linearly dependent on tension. Calcium preferentially potentiated the relaxation, and apparently reduced the resting tension in the elastic element. The calcium site appears specific for calcium, as other divalent cations inhibited its action. Dihydrostreptomycin inhibited the positive rate, but its effect could not be explained by a simple channel block, and it seems inconsistent with screening of negative charge in the mouth of the transduction channel. PMID- 2555461 TI - Interaction of phencyclidine with voltage-dependent potassium channels in cultured rat hippocampal neurons: comparison with block of the NMDA receptor ionophore complex. AB - Whole-cell voltage-clamp recording techniques were used to investigate the blockade of voltage-dependent K+ channels by phencyclidine (PCP) in cultured rat hippocampal neurons. All recordings were carried out in the presence of tetrodotoxin (1-2 microM) to eliminate Na+ currents. Step depolarization from a holding potential of -40 mV activated a slowly rising, minimally inactivating K+ current (IK). PCP (0.5-1000 microM) caused a reduction in the maximum conductance of IK [IC50(+30 mV), 22 microM] without altering its voltage dependency. The PCP block of IK diminished at depolarized potentials. Analysis according to the scheme of Woodhull (1973) suggested that block occurs via binding to an acceptor site (presumably within the channel pore) that senses 40-50% of the transmembrane electrostatic field. PCP had no effect on the kinetic properties of IK and the block failed to show use dependency, suggesting that PCP may bind to the IK channel via a hydrophobic mechanism not requiring open channels. For comparison, we also investigated the effect of PCP on the transient K+ current, IA, activated by step depolarization following a 200 msec prepulse to -90 mV (20 mM tetraethylammonium was present in the bathing solution to reduce IK). In contrast to the potent blocking action of PCP on IK, the drug only affected IA at high concentrations [IC50(+30 mV), 224 microM]. At concentrations causing substantial block (300-500 microM), PCP produced an acceleration in the IA inactivation rate, and, for brief (5-6 msec) depolarizing steps, the suppression of IA was use dependent. These observations suggest that PCP block of IA requires open channels. PCP reduced inward current responses induced by the excitatory amino acid agonist N-methyl-D-aspartate (NMDA) at substantially lower concentrations than those required for its effects on K+ channels [IC50(-60 mV), 0.45 microM]. The PCP-like dioxadrol stereoisomer dexoxadrol (10 microM) blocked NMDA-evoked inward current responses, while its behaviorally inactive enantiomer levoxadrol did not. Dexoxadrol and levoxadrol also blocked IK in a stereoselective fashion (IC50's, 73 and 260 microM, respectively), whereas the sigma ligands (+)- and (-) SKF 10,047 and (+)-3-[3-hydroxyphenyl]-N-(1-propyl)piperidine [(+)-3-PPP] had little effect on the current (IC50's, greater than 300-500 microM). We conclude that PCP causes a selective, voltage-dependent block of IK in hippocampal neurons via a PCP- and not a sigma-type acceptor site.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2555462 TI - Nonlinearity and facilitation in phosphoinositide signaling studied by the use of caged inositol trisphosphate in Xenopus oocytes. AB - The phosphoinositide signaling pathway, which mediates neurotransmitter responses, was studied in Xenopus oocytes by recording membrane currents evoked using lightflash photolysis of caged inositol trisphosphate (caged IP3) to produce rapid and reproducible transients in intracellular IP3 levels. Photolysis of caged IP3 evoked currents which were carried largely by chloride ions and depended upon intracellular, but not extracellular, calcium. A given light flash evoked larger responses when the amount of caged IP3 loaded into the oocyte was increased, and illumination of the vegetal hemisphere gave larger responses than the animal. Long (10 sec) light exposures produced oscillatory currents, resembling responses to serotonin and other agonists, which became larger, more transient, and of shorter latency as the light intensity was increased. Brief (ca. 100 msec) flashes evoked a single "spike" of current. The caged IP3 response showed a threshold, in that light flashes had to be greater than a certain intensity and duration before currents could be detected. Associated with this, sub- and suprathreshold light flashes caused a long-lasting (seconds or minutes) potentiation of responses to subsequent test flashes. The lightflash response was also potentiated by a preceding intracellular injection of IP3 and by extracellular application of an agonist thought to induce IP3 liberation. However, intracellular injections of calcium depressed the response. We conclude that the liberation of calcium from intracellular stores varies nonlinearly with the intracellular level of IP3. This phenomenon may explain earlier observations, including the long latency of currents evoked by low doses of agonists such as acetylcholine and serotonin, and the nonlinear facilitation seen between these agonists. Further, it suggests a mechanism for "chemical integration," which may be important in the functioning of neurons and other cells which use IP3 as an intracellular messenger. PMID- 2555463 TI - A controlled study of efficacy of interstitial or external irradiation in a virus induced brain-tumor model in rats. AB - In a controlled study of interstitial radiotherapy in the avian sarcoma virus (ASV)-induced glioma model in rats, prolongation of survival was demonstrated (p = 0.08 in Experiment 1 and p = 0.03 in Experiment 2) following mean dosages of 7582 to 9902 cGy 125I, when compared to nontreatment or to control studies with implantation of nonradioactive seeds. More significant (p = 0.02) prolongation of survival was demonstrated following external beam whole-head radiotherapy with nine fractions of 333 cGy, three times weekly over 3 weeks (total dose 3000 cGy). Survival was more prolonged when whole-head radiotherapy was begun 35 days following virus inoculation rather than at 71 days, probably reflecting a greater efficacy with smaller tumor targets. PMID- 2555464 TI - Technetium-99m (V) DMSA uptake in amyloidosis. AB - Technetium-99m(V) DMSA scintigraphy was performed in two patients with pathologically confirmed amyloidosis associated with plasmacytoma. Significant uptake of the tracer was found in the deposition of amyloid. Technetium-99m(V) DMSA scintigraphy could be useful in determining the appropriate region of biopsy and in forecasting the prognosis of patient with plasmacytoma. PMID- 2555465 TI - Interrelated effects of dietary fiber and fat on lymphatic cholesterol and triglyceride absorption in rats. AB - Lymph cannulated rats were administered intragastrically a test emulsion containing 25 mg of [14C]cholesterol, 50 mg of either guar gum, cellulose or chitosan, and 200 mg of either safflower, high-oleic safflower or palm oil, and the absorption of labeled cholesterol and fatty acids was measured. The type of both dietary fiber (P less than 0.001) and fat (P less than 0.05) significantly influenced cholesterol absorption. A significant interaction of fiber and fat on cholesterol absorption (P less than 0.05) was also observed. Chitosan effectively lowered cholesterol absorption more than did guar gum or cellulose, and this effect was more significant when given with safflower or high-oleic safflower oil than with palm oil. When guar gum was the source of dietary fiber, dietary fats did not modify cholesterol absorption. Dietary fiber also significantly affected triglyceride absorption (P less than 0.05). Absorption tended to be low in the chitosan, high in the cellulose and intermediate in the guar gum group. Absorption of safflower and high-oleic safflower oils tended to be higher than that of palm oil when cellulose or guar gum was fed. Guar gum, as compared with the other fibers, altered the absorption pattern of both cholesterol and triglyceride. The results showed that the type of dietary fat significantly influenced the effect that dietary fiber exerted on lipid absorption. PMID- 2555466 TI - Neomycin has no persistent sparing effect on vitamin B-12 status in pectin-fed rats. AB - In the present study, rats were depleted of vitamin B-12 with fiber-free or 5% pectin diets, with or without neomycin. Through use of this intestinal antibiotic reported to "spare" vitamin B-12, we sought to determine if bacterial fermentation of pectin might explain our previous observations of negative effects of pectin on vitamin B-12 status. However, neomycin did not lessen interference by pectin with vitamin B-12 metabolism. Pectin increased urinary methylmalonate and decreased propionate oxidation to a greater extent in the presence than in the absence of neomycin. Also, regardless of the presence of neomycin, the biologic half-life of injected [57Co]vitamin B-12 was 58 d for rats fed the fiber-free diets and only 38 d for rats fed 5% pectin diets. Neomycin delayed early fecal excretion of 57Co but had no persistent effect. Thus, neomycin-sensitive bacteria do not mediate the negative effects of pectin on vitamin B-12 status. Pectin may interfere directly with vitamin B-12 absorption or may stimulate vitamin B-12 uptake or propionate production by microbial species that have adapted to neomycin. PMID- 2555467 TI - Comparison of the increment in plasma eicosapentaenoate concentrations by fish oil intake between young and middle-aged volunteers. AB - The effect of age on eicosapentaenoic acid (20:5 n-3; EPA) incorporation into plasma lipids was investigated in young volunteers (8 males, 19 +/- 1 yr) and middle-aged volunteers (6 males, 53 +/- 7 yr). They were asked to take 5.4 g fish oil per day for one week. The increment in EPA in the cholesteryl ester fraction after the supplementation was significantly greater in the middle-aged group (delta = 1.69%) than in the young group (delta = 0.44%) (p less than 0.05). The food intake analyzed for 3 consecutive days just before the supplementation revealed that the young group took more linoleate (17 vs. 10 g) than the middle aged group. There was a significant inverse correlation between the increment in EPA in the cholesteryl ester fraction after the supplementation and daily linoleate intake among all the volunteers combined (r = -0.63, p less than 0.02). The higher increment in EPA in cholesteryl ester in the middle-aged group might be due to less intake of linoleate and not due to the difference in age itself. PMID- 2555468 TI - Tissue response to facial contour augmentation with dense and porous hydroxylapatite in rhesus monkeys. AB - Using extraoral incisions, subperiosteal pockets were created bilaterally over the zygomatic and mandibular areas in six rhesus monkeys. One side of each anatomic site received a dense block of hydroxylapatite (HA) implant, and the contralateral side received an equivalent-sized block of porous HA implant. Monkeys were killed at 3, 6, and 12 months, and the implants were manually tested for mobility. The implants were then retrieved in block specimens. Half of each specimen was decalcified, embedded in paraffin, and stained. The other half was embedded in plastic, and sections were stained or carbon coated for scanning electron microscope histometry. The dense HA implants showed complete fibrous encapsulation and they popped out when cut in half. The porous HA implants were attached to the underlying cortex by bone ingrowth. The porous implant volume sampled within 2.5 mm of underlying cortex contained 33.0% HA matrix and 35.7% bone. The surface area of the porous HA matrix (4.8 mm2/mm3) was 54.3% covered by bone ingrowth. It is hypothesized that micromotion may have accounted for the lack of osseointegration of the dense HA specimens. In contrast, the early ingrowth of fibrous tissue into the porous HA block might be responsible for reducing micromotion to levels acceptable for ingrowth of bone. The contrasting biologic response of implant sites to two variants of the same implant material supports the value of comparative studies to permit informed surgical selection decisions. PMID- 2555469 TI - Invasive Aspergillus infection in chronic granulomatous disease: treatment with itraconazole. PMID- 2555470 TI - Use of quinolones in childhood. PMID- 2555471 TI - Seroepidemiologic studies of cytomegalovirus and Epstein-Barr virus infections in relation to human immunodeficiency virus type 1 infection in selected recipient populations. Transfusion Safety Study Group. AB - Antibodies to human cytomegalovirus (CMV) and Epstein-Barr virus (EBV) were evaluated among 1,171 persons with and without antibodies to human immunodeficiency virus type 1 (anti-HIV-1). These included 97 blood donors, 577 persons given blood components or products, and 497 controls. A significantly higher proportion of anti-HIV-1 positive than -negative donors were anti-CMV positive, a finding associated with homosexual contact among some of the former. Among subjects with treated clotting disorders, there was no difference in prevalence of anti-CMV or anti-EBV between anti-HIV-1-positive and -negative persons. The prevalence of antibodies to EBV early antigens showed no relationship to anti-HIV-1 status. Anti-CMV positivity in anti-HIV-1-negative donors was associated with an increase in mean CD8 counts and lower mean CD4/CD8 ratio. Anti-CMV and anti-EBV positivity in anti-HIV-1-positive subjects with treated clotting disorders was not associated with a lower CD4 or higher CD8 count than HIV-1 infection alone. Subjects who developed AIDS after enrollment had no significant difference in median time from entry to diagnosis when analyzed by serologic evidence of CMV and EBV antibody status at entry, and a few subjects had AIDS at entry without serologic evidence of prior CMV or EBV infection. The overall results are consistent with acquisition and progression of HIV-1 independently of coincident CMV or EBV infection. PMID- 2555472 TI - Infraclinical neuropathies related to immunodeficiency virus infection associated with higher T-helper cell count. AB - To assess subclinical involvement of the peripheral nervous system and its relationship to the immunological status of human immunodeficiency virus (HIV) infected patients, we prospectively studied the peripheral nerve conductions and the subsets of peripheral blood lymphocytes of HIV patients. Fifty-seven patients, aged 20-54 years, 28 homosexuals and 29 heterosexuals, classified as CDC II-III (40 patients) and CDC IV (17 patients) were studied. No neurological symptoms or signs were present on bedside examination or questioning for all of the CDC II-III patients. For each conduction measured (H-reflex, sural and sciatic velocities, sural amplitude), the geometric mean was below normal values (p less than 0.05). Among the 57 HIV-infected patients, 20 (35%) had a significant decrease of at least one electrophysiologic measurement: 10/17 CDC IV patients vs. 10/40 CDC II-III patients (p less than 0.05). Two or more simultaneous abnormalities were more often observed in the CDC IV than in the CDC II-III group (9/10 vs. 4/10) (p less than 0.01). In the CDC II-III group, patients with subclinical neuropathies had higher T-helper and lower T-suppressor cell counts and higher T-helper/T-suppressor ratios than random patients without any neurologic abnormalities (1,057 vs. 657 cells/microliters, p less than 0.05, and 757 vs. 1,304 cells/microliters, p less than 0.01, 1.55 vs. 0.68, p less than 0.01, respectively). We conclude that (a) the peripheral nervous system is precociously and subclinically involved in the HIV disease; (b) the patients with infraclinical neuropathies have a significantly higher T-helper cell count than those without any neurologic involvement. The mechanism and the prognostic value of these two findings remain to be more precisely examined, and further studies are required. PMID- 2555473 TI - Cervical and vaginal squamous cell abnormalities in women infected with human immunodeficiency virus. AB - We sought to determine whether women infected with human immunodeficiency virus (HIV) had cervicovaginal cellular changes suggesting lower genital tract neoplasia or human papillomavirus (HPV) infection at a rate different from that in women without HIV infection. In a blinded fashion, cytological preparations of cervicovaginal smears from women infected with the HIV were analyzed and compared to preparations from women at high risk for but not infected with HIV. Eleven of 35 (31%) HIV-infected subjects had evidence of squamous abnormalities compared with 1 of 23 (4%) non-HIV-infected women (p = 0.019). Nine of 35 (26%) HIV infected women had cytohistological evidence of human papillomavirus (HPV) infection compared to 1 of 23 (4%) non-HIV-infected women (p = 0.072). We conclude that HIV-infected women have a high prevalence of cervical and vaginal cytological abnormalities and evidence of genital HPV infection. Further study is necessary to determine whether there is an increased risk for cervicovaginal neoplastic disorders in women infected with HIV. PMID- 2555474 TI - The effect of extracellular weak acids and bases on the intracellular buffering power of snail neurones. AB - 1. Intracellular pH (pHi) was measured in snail neurones using pH-sensitive glass microelectrodes. The influence of externally applied weak acids and bases on the total intracellular buffering power (beta T) was investigated by monitoring the pHi changes caused by the intracellular ionophoretic injection of HCl. 2. In the absence of weak acids or bases a reduction in the extracellular HEPES concentration had no effect on pHi or on beta T. It did, however, reduce slightly the rate of pHi recovery following HCl injection. 3. The presence of CO2 greatly increased beta T. However, as predicted for an open buffer system, the contributions to intracellular buffering by CO2 (beta CO2) decreased as pHi decreased. 4. When added to the superfusate, procaine, 4-aminopyridine, trimethylamine and NH4Cl (1-10 mM) all increased steady-state pHi. Procaine was fastest at increasing pHi and 4-aminopyridine the slowest. All four of these weak bases increased beta T. 5. The intracellular buffering action by these weak bases varied. HCl injection in the presence of procaine usually resulted in steady state pHi changes with no pHi transients. In the presence of the other three weak bases HCl injections resulted in intracellular acidifications which were followed by pHi recovery-like transients. However, these were not blocked by SITS (4 acetamido-4'-isothiocyanatostilbene-2,2'-disulphonic acid) or by CaCl2 and I thus conclude that these transients were as a result of slow or incomplete intracellular buffering by the weak bases. 6. In many cells there was a good correlation between the measured contributions to intracellular buffering by the weak bases (beta base) and those predicted assuming a simple two-compartment open system. In all cases, as predicted, beta base increased as pHi decreased. 7. I found a clear relationship between the concentration of external buffer (HEPES) and the rate at which weak bases, applied to the superfusate, were able to increase pHi. The greater the extracellular buffer concentration the greater was the speed of intracellular alkalinization. 8. Lowering the extracellular buffer concentration reduced the efficiency of intracellular buffering by weak bases in response to an intracellular acid load. HCl injection in the presence of weak base caused a larger initial intracellular acidification if the extracellular HEPES concentration was reduced. 9. In conclusion, both weak acids and weak bases can make very large, pHi-dependent contributions to intracellular buffering by way of open buffer systems.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2555475 TI - Potentiation and suppression by eserine of muscarinic synaptic transmission in the guinea-pig hippocampal slice. AB - 1. The effects of the anticholinesterase eserine on CA3 pyramidal cells and dentate gyrus granule cells in guinea-pig hippocampal slices were investigated with single-electrode current-clamp and voltage-clamp recording. 2. In the majority of cells superfused with eserine (0.5-10 microM) for 3-10 min, tetanic stimulation near the cell layers elicited a delayed depolarization (slow EPSP; duration up to 60 s) at a pre-stimulation membrane potential of -60 mV. The slow EPSP was blocked by atropine (1 microM). 3. Under voltage clamp at -60 mV holding potential, an apparent inward current (slow EPSC) with a similar time course to the slow EPSP was observed. 4. The amplitude of the delayed inward current was about 50 pA. The amplitude increased at holding potentials more positive than -60 mV. At holding potentials negative to -60 mV, the delayed inward current was too small to allow reliable analysis. In the absence of eserine, there was a delayed inward current, which was rather small, however, due to a superimposed outward current. 5. Eserine reduced the after-hyperpolarization following a train of action potentials. This effect was antagonized by atropine, but not to pirenzepine. In voltage-clamp recording, eserine reduced a current termed IAHP. 6. CA3 neurones treated with eserine exhibited a region of negative slope conductance (in tetrodotoxin). The slow inward current which developed at clamp potentials between -50 and -40 mV was reduced by Ni2+ (50 microM). The effect of eserine on slope conductance increased with time of exposure. In all neurones superfused with eserine for more than 60 min, burst discharges were observed. Burst discharges were blocked by atropine and Ni2+, but not by pirenzepine. 7. In cells superfused with eserine for more than 1 h, tetanic stimulation failed to elicit a slow EPSP or EPSC. Currents induced by focal acetylcholine (ACh) application were first enhanced by eserine, but blocked after exposure to eserine for more than 1 h. Blockade of ACh-induced currents was also observed after bath application of carbachol (CCh) in a concentration (0.2 microM) in which it did not induce an inward current at -60 mV holding potential. Further, the slow EPSP faded when elicited by repeated tetanic stimulation. 8. While the observed effects of eserine on hippocampal neurones can be explained by eserine's well known ability to block acetylcholinesterase activity, our data indicate that the effects of eserine involve more than one muscarinic receptor site, i.e. desensitizing and non-desensitizing postsynaptic receptor sites. PMID- 2555476 TI - Properties of the block of single Na+ channels in guinea-pig ventricular myocytes by the local anaesthetic penticainide. AB - 1. The blocking mechanism of a disopyramide derivative Penticainide (2-alkyl-(4 (dialkylamino)-2-)pyridyl-butyramide) on cardiac Na+ channels has been studied using single-channel analysis in cell-attached and inside-out patches from guinea pig ventricular cells. Penticainide was applied in concentrations between 3 and 100 microM. The S-enantiomer of DPI 201-106 (5 microM) was used as a tool to slow the inactivation, improve the time resolution by prolonging the mean open time, and to increase the number of openings per depolarization of the channel. 2. When in cell-attached or inside-out patch experiments up to 100 microM-Penticainide was applied to the bathing solution no significant effect was observed on the probability of the channel being open or on the mean open time. 3. In cell attached patch experiments with 100 microM-Penticainide in the pipette, the open state probability of the Na+ channel was much lower than in the absence of Penticainide. No significant changes were found in the potential of half-maximum activation or in the slope of the activation curve. The maximum open-state probability was reduced by a factor five in the presence of Penticainide. The single-channel conductance was not affected by the drug. 4. The decrease in the probability of the channel being open was mainly due to an increased probability of observing sweeps with no activity ('nulls'). 5. A dramatic relief from the block was observed when pauses were interposed into the normal activation pattern, or when the pacing rate was reduced. 6. The distribution of the open times of the bursting Na+ channel could be fitted with two exponentials. Penticainide in the patch pipette reduced the mean open time. Also the contribution of the number of long openings to the total number of openings was reduced. 7. Closed-time distribution was also fitted with two exponentials. Penticainide in the patch pipette prolonged the long mean closed time. The contribution of the number of short closings to the total number of closings was decreased. 8. Penticainide in the patch pipette did not significantly change the time constant of the decay of the ensemble-averaged currents measured at -30 mV. Because of the close correlation between the burst duration and the time constant of inactivation (Nilius, Vereecke & Carmeliet, 1988b), we conclude that no striking effect of Penticainide on the burst duration can be expected.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2555478 TI - Oral hairy leukoplakia: observations in 95 cases and review of the literature. AB - Oral hairy leukoplakia (HL) was observed in 25.4% of 373 HIV-seropositive patients (n = 95). 87 were men of an average age of 37.1 yr, 8 were women (30.3 yr). 71.6% of the patients were male homosexuals. At initial diagnosis of HL the majority of cases was classified as CDC IVc1 (45.3%) and CDC II (22.1%). Average CD4/CD8 ratio (n = 19) was 0.24 with a range of 0.04-1.0. Thirty biopsies of HL revealed some of the histologic features thought to be characteristic. In only 20 of 30 biopsies EB-virus-specific-capsid antigen was detected. The problems of clinical and histological diagnosis of HL are discussed. Further strict criteria are necessary in order to define HL more distinctly. PMID- 2555477 TI - Adrenal responses to calcitonin gene-related peptide in conscious hypophysectomized calves. AB - 1. Right adrenal and various cardiovascular responses to an intra-aortic infusion of calcitonin gene-related peptide (CGRP; 4 micrograms min-1) have been investigated in the presence and absence of exogenous adrenocorticotrophin ACTH1 24 (2 or 5 ng min-1 kg-1, I.V.). The adrenal clamp technique was employed in conscious calves in which the pituitary stalk had been cauterized 3-7 days previously. 2. At the higher dose (5 ng min-1 kg-1) the I.V. infusion of ACTH raised mean plasma ACTH concentration by about 1000 pg ml-1 and mean right adrenal cortisol output by about 750 ng min-1 kg-1. Under these conditions the intra-aortic infusion of CGRP had no apparent effect on adrenal cortisol output by about 750 ng min-1 kg-1. Under these conditions the intra-aortic infusion of CGRP had no apparent effect on adrenal function, other than to produce moderate adrenal vasodilatation. In contrast, in the absence of exogenous ACTH, the same dose of CGRP produced a substantial rise in cortisol output, which rose steadily to a peak mean value of 409 +/- 31 pg min-1 kg-1 at 10 min. It also significantly inhibited the release of free, but not of total, met5-enkephalin-like immunoreactivity from the gland (P less than 0.001) together with a significantly greater fall in adrenal vascular resistance (P less than 0.001). 3. At the lower dose of ACTH (2 ng min-1 kg-1, I.V.) CGRP raised mean plasma cortisol output from 314 +/- 31 to 486 +/- 44 ng min-1 kg-1 (P less than 0.01) and this effect was not attributable to an increase in the adrenal presentation rate of ACTH. 4. It is concluded that this peptide exerts a steroidogenic action on the adrenal cortex which is manifest in the absence of exogenous ACTH in the functionally hypophysectomized calf. PMID- 2555479 TI - In situ detection of human papillomavirus types 13 and 32 in focal epithelial hyperplasia of the oral mucosa. AB - 17 cases of focal epithelial hyperplasia of the oral mucosa (FEH, Heck's disease) were investigated for the presence of human papillomavirus (HPV) nucleic acid sequences by means of in situ DNA hybridization using biotinylated DNA probes of HPV types 1, 6, 11, 13, 16, 18, and 32. Ten of 17 cases were positive for HPV 13 DNA in contrast to 6 of 17 positive cases obtained after application of the HPV 32 probe, with a double infection in one case. The results of our study suggest, that HPV 13 and HPV 32 are very specifically found in lesions of FEH and can be detected in a high percentage of cases using in situ hybridization. PMID- 2555480 TI - Systematized procedure of crown preparation. AB - The theoretical considerations of the marginal fit of crowns are discussed and the conventional approach in tooth-cutting procedures is addressed. Gingival marginal surface irregularities described as "peak and valley" and "stepping" effects can leave a cement line in excess of 80 microns after cementation. A regimented method of tooth reduction for metal ceramic crowns, using specially designed rotary cutting instruments, is described. The suggested technique accomplishes the crown preparation more rapidly with a consistent standard of excellence. PMID- 2555481 TI - Leakage of chemical and light-cured basing materials. AB - This study investigated the sealing properties of chemical and light-cured basing materials in dentin. Fifty extracted molars were selected and cleaned. Slot preparations with standard dimensions were made. The prepared cavities were filled with Cavitec, Life, Ketac-Bond, VLC Dycal, and Cavalite materials. Samples were immersed in 0.05% crystal violet solution for 2 hours. The samples were then embedded in clear casting resin, sectioned, and examined with a microscope. Results indicated that Cavalite and Ketac-Bond materials allow a greater degree of dye penetration that the three other materials. Cavitec, VLC Dycal, and Life materials were similar to each other in degree of dye penetration. PMID- 2555482 TI - Amplification of small signals by voltage-gated sodium channels in drone photoreceptors. AB - Photoreceptor cells of the drone, Apis mellifera male, have a voltage-gated Na+ membrane conductance that can be blocked by tetrodotoxin (TTX) and generates an action potential on abrupt depolarization: an action potential is triggered by the rising phase of a receptor potential evoked by an intense light flash (Autrum and von Zwehl 1964; Baumann 1968). We measured the intracellular voltage response to a small (9%), brief (30 ms) decrease in light intensity from a background, and found that its amplitude was decreased by 1 microM TTX. The response amplitude was maximal when the background intensity depolarized the cell to -38 mV. With intensities depolarizing the cell membrane to -45 to -33 mV the average response amplitude was decreased by TTX from 1.2 mV to 0.5 mV. TTX is also known to decrease the voltage noise during steady illumination (Ferraro et al. 1983) but, despite this, the ratio of peak-to-peak signal to noise was, on average, decreased by TTX. The results suggest that drone photoreceptors use voltage-gated Na+ channels for graded amplification of responses to small, rapid changes in light intensity. PMID- 2555484 TI - Human immunodeficiency virus infection in a Dublin general practice. AB - A group general practice in Dublin's inner city has had extensive experience of intravenous drug users since the late 1970s. Since 1985 a total of 54 human immunodeficiency virus (HIV) seropositive patients have attended the practice, of whom 48 are intravenous drug users, four are the children of drug users and two have been infected through sexual contacts. Three patients have developed the acquired immune deficiency syndrome and at least eight have symptomatic HIV disease. Sixty per cent of Ireland's seropositive population have been infected through intravenous drug abuse but nationally only 16% of all intravenous drug users tested are seropositive; in the study practice, however, at least 35% (48/137) of known intravenous drug users are seropositive. PMID- 2555483 TI - [Vagal paraganglioma associated with a rare malformation of the aortic arch. Apropos of a case and review of the literature]. AB - The authors report about a very peculiar case of cervical paraganglioma which had two specific features. This is a familial case with 2 sisters presenting with bilateral paragangliomas. In addition, this patient's aortic trunk is very peculiar, with a retroesophageal right subclavian artery and, above all, a divided intrathoracic right common carotid artery, producing an ascending pharyngeal supply for the right paraganglioma, which can be embolized without any risks of reflux. PMID- 2555485 TI - The general practitioner and human immunodeficiency virus infection: an insight into patients' attitudes. AB - In a study of 100 patients with the acquired immune deficiency syndrome (AIDS), 77% were registered with a general practitioner and a further 14% wished to register. Of those 77 who were registered with a general practitioner, only 47 doctors knew their diagnosis; 19 of the 77 did not want their general practitioner to know. Of this small group of 19, a proportion would visit their general practitioner with symptoms, some of which may be related to AIDS. The main difficulty for patients in telling a general practitioner about their illness was a perceived lack of confidentiality and lack of sympathy. Patients valued understanding and expertise as most important in a general practitioner. This study provides an analysis of why general practice is not seen as a significant resource for many patients with AIDS in the London area and suggests some initiatives to enhance the appropriate use of primary care services. PMID- 2555486 TI - The Worcester development project: general practitioner satisfaction with a new community psychiatric service. AB - General practitioners in the Worcester development project area were interviewed about their experience of using the new community based psychiatric services. Of those who remembered the previous asylum based services most thought the new services were an improvement in many respects and were satisfied with the care provided for their patients. However, there were difficulties in obtaining emergency admissions, and criticisms of the social work service being slow to respond to requests for help. General dissatisfaction was expressed about feedback, particularly from the community psychiatric nursing service. General practitioners are expected to have a key role in the coordination of community services. However, most general practitioners interviewed had no particular interest or training in psychiatry. This, coupled with the inadequate information they received and the possibility of their workload increasing as more patients move out of hospital, raises questions about how they may be helped to fulfil this coordinating role in community psychiatric services. PMID- 2555487 TI - A classification of prescription errors. AB - Three independent methods of study of prescription errors led to the development of a classification of errors based on the potential effects and inconvenience to patients, pharmacists and doctors. Four types of error are described: type A (potentially serious to patient); type B (major nuisance - pharmacist/doctor contact required); type C (minor nuisance - pharmacist must use professional judgement); and type D (trivial). The types of frequency of errors are detailed for a group of eight principals from one health centre. There were a total of 504 errors from 15,916 prescription items (3.17%) during a three month observation period. A close correspondence was found between individual doctor's types of error rates, suggesting that doctors who make type C and D errors are also likely to make type B (major nuisance) errors. A system of feedback of errors from each doctor was devised. No significant reduction was seen in error rates, possibly because the group of self selected doctors taking part had low error rates initially. It is suggested that pharmacists and doctors should work closely together to prevent the potentially harmful consequences of prescription errors. PMID- 2555488 TI - Hazards of scoop measurements in infant feeding. AB - Mothers attending infant welfare clinics were asked to measure milk powder from a standard packet with the scoop provided by the manufacturers. Wide variations were found in the weight of powder obtained, with the highest scoop weight (5.6 g) being double the lowest (2.8 g). It is suggested that the scoop method of measuring milk powder is so inaccurate that the manufacturers should present their product in small pre-measured packets. PMID- 2555489 TI - Paying general practitioners: shedding light on the review of health services. AB - This paper reviews evidence from recent research on the effects of different methods of remunerating general practitioners. Each method is examined in terms of patient use of health services in general, use of services by different groups in society and health outcome. Little is known about the effects of capitation as it currently exists in the UK, salaries or special payments for 'good practice', although evidence from British research is likely to be forthcoming on the last of these. Both health maintenance organizations and charges deter utilization, although little is known about the effect of this reduced demand. Furthermore, these two methods of financing health care appear to discriminate between members of society on lower and higher incomes in terms of both service use and health outcome. Fees for items of service provided tend to lead to unnecessary demands for fee yielding services by patients on the recommendation of their doctors. Although more evidence on different methods of remuneration is required, the importance of what is already known depends on the objectives of health care provision. PMID- 2555491 TI - Drug information for family doctors--is an informal style acceptable? PMID- 2555490 TI - Can general practitioners counsel? AB - It has been suggested that general practitioners are in a prime position to counsel patients presenting with psychosocial problems. While many doctors use counselling skills in their consultations few have received training in counselling and the difference between the use of counselling skills and the process of counselling is not always understood. This paper examines the differences between counselling and counselling skills and compares the role of doctors and counsellors. It is concluded that there is a need for trained counsellors to work alongside general practitioners and that this is of benefit to patients and all members of the primary care team. PMID- 2555492 TI - The advertising debate. PMID- 2555493 TI - Community care for the elderly. PMID- 2555494 TI - Crohn's disease. PMID- 2555495 TI - Chronic pelvic pain. PMID- 2555496 TI - Caring for the mentally handicapped in the community. PMID- 2555497 TI - AIDS and the future general practitioner. PMID- 2555498 TI - Ultra nappy rash. PMID- 2555499 TI - Working for patients--a journey into the unknown. PMID- 2555501 TI - Drug defect reporting. PMID- 2555500 TI - Quality of care in general practice--lessons from the past. PMID- 2555503 TI - Analysis of the hospital experience completed by general practitioner trainees in 1984-87. AB - This report gives details of the hospital experience undertaken by doctors who have completed vocational training for general practice and is based on the information presented by applicants for the certificate of the Joint Committee on Postgraduate Training for General Practice. PMID- 2555502 TI - Women doctors' career choice and commitment to medicine: implications for general practice. AB - This study examined the work experiences and plans of a national sample of 150 female medical graduates of 1976, 1980 and 1984. The sample was exhaustively traced and information obtained about 97% of the doctors, including 100% of the doctors ever likely to practise in the UK. The findings show a high recent and planned participation rate in medical practice, especially general practice, among these women graduates and no involuntary unemployment. Increased numbers of women at medical school will result in manpower changes, particularly in general practice, but these increases will not counter possible overproduction of medical graduates. The study also demonstrates that it is possible to achieve a high response rate among medical graduates by using a telephone interview. PMID- 2555504 TI - Words or numbers? The evaluation of probability expressions in general practice. AB - A sample of 56 general practitioners were asked to rate, on a percentage scale, 23 words or phrases which denote frequency or likelihood. The hypothetical context of the exercise was that of communicating to patients the probability of a side-effect (headache) arising from an unspecified prescription medicine. Median phrase ratings ranged from 'never' at 0% to 'certain' at 95% with a 50% rating given to the phrase 'reasonable chance'. Despite relatively large variance in ratings between respondents, the median ratings of a number of phrases were similar, and some identical, to other studies from different medical professionals. Although the clinical context in which a given expression of probability is used may affect its meaning, the results are encouraging and suggest that phrases denoting likelihood might be systematically codified to enhance communication between doctor and patient. To move towards this objective more research is needed to evaluate how patients interpret expressions of probability, and the relative effectiveness of different modes of communicating likelihood. PMID- 2555505 TI - High titer of antibody to the Epstein-Barr virus membrane antigen in sera from patients with rheumatoid arthritis and systemic lupus erythematosus. AB - Sera from patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) contained more antibody to the Epstein-Barr virus membrane antigen (EBV MA) than sera from healthy controls. Since antibody titer to EBV MA closely correlates with viral neutralization, it was inferred that these patients were frequently exposed to infectious EBV, producing high titers of neutralizing antibody. PMID- 2555506 TI - Insulinoma. PMID- 2555507 TI - Cholecystectomy, duodeno-gastric reflux and polyposis. PMID- 2555508 TI - Hypoglycaemia for 26 years due to an insulinoma. PMID- 2555509 TI - Cytomegalovirus (CMV) enterocolitis. PMID- 2555510 TI - Specific inhibition of benzodiazepine receptor binding by some N-(indol-3 ylglyoxylyl)amino acid derivatives: stereoselective interactions. AB - Several optically active N-(indol-3-ylglyoxylyl)amino acid derivatives were synthesized and tested for [3H]flunitrazepam displacing activity in bovine brain membranes. IC50 values were measured and revealed that the D form of the amino acid moiety of the compounds was more potent than both the L form and racemic form, suggesting a key role of the amino acid stereochemistry on the affinity to the benzodiazepine receptors. GABA ratio and proconvulsant/convulsant data reported for the most active compounds reveal they behave as inverse agonists at the benzodiazepine receptor. PMID- 2555511 TI - Selective elimination of interactions: a method for assessing thermodynamic contributions to ligand binding with application to rhinovirus antivirals. AB - A new method for evaluating the free energy of various physical interactions, such as hydrogen-bond, electrostatic, or van der Waals interactions, is presented. Rather than destroying or creating whole groups, selective (pairwise) interactions are eliminated from the total potential energy and the energy difference with the fully interacting system is evaluated. The exponential ensemble average of such an energy difference is then directly related to the corresponding free energy difference. This procedure is then applied to a rather large protein-ligand system involving the coat proteins of a human rhinovirus and an antiviral ligand. The results seem to indicate that a particular bent hydrogen bond between the ligand and protein system may not be favorable for binding. The method presented gives an estimate of the hydrogen bond free energy contribution with an available trajectory that was previously computed without the expenditure of sizeable computational resources such as recomputing a trajectory. This procedure is effective and efficient for computing the free energy for a given type of physical interaction. It can be used for calculating the binding energy differences for various interactions which can be used to guide the search for isosoluble synthetic targets. PMID- 2555512 TI - Potent and prolonged acting cyclic lactam analogues of alpha-melanotropin: design based on molecular dynamics. AB - Utilizing results from previous structure-activity relationships and theoretical studies of alpha-melanotropin (alpha-MSH, Ac-Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp Gly-Lys-Pro-Val-NH2) and its related superpotent analogues, Ac-[Nle4,D-Phe7] alpha-MSH and Ac-[Cys4,Cys10]-alpha-MSH, we have designed a new class of alpha MSH4-13 and alpha-MSH4-10 cyclic lactam fragment analogues of alpha-melanotropin. The cyclic peptides have the following general structures: Ac-[Nle4,Xxx5,D Phe7,Yyy10,Gly11]-alpha-MSH4-13- NH2 and Ac-[Nle4,Xxx5,D-Phe7,Yyy10]-alpha-MSH4 10-NH2, where Xxx = Glu or Asp and Yyy = Lys, Orn, Dab, or Dpr. Formation of the lactam bridge between the side-chain groups Xxx and Yyy was performed either in solution or on a solid-phase support. Seven cyclic peptides were prepared and bioassayed for their melanotropic potency by using standard frog (Rana pipiens) and lizard (Anolis carolinensis) skin bioassays. Relative to alpha-MSH (relative potency = 1), the potencies of the cyclic peptides in the lizard skin bioassay were as follows: alpha-MSH (1); Ac-[Nle4,Glu5,D-Phe7,Lys10,Gly11]-alpha-MSH4-13- NH2 (6); Ac-[Nle4,Asp5,D-Phe7,Lys10,Gly11]-alpha-MSH4-13- NH2 (100); Ac [Nle4,Glu5,D-Phe7,Lys10]-alpha-MSH4-10-NH2 (9); Ac-[Nle4,Asp5,D-Phe7,Lys10]-alpha MSH4-10-NH2 (90); Ac-[Nle4,Asp5,D-Phe7,Orn10]-alpha-MSH4-10-NH2 (20); Ac [Nle4,Asp5,D-Phe7,Dab10]-alpha-MSH4-10-NH2 (5); Ac-[Nle4,Asp5,D-Phe7,Dpr10]-alpha MSH4-10-NH2 (5). Similar results were obtained in the frog skin bioassay, but the analogues were much less potent. Cyclic melanotropins with 23-membered rings exhibited 100-fold higher melanotropic potency than alpha-MSH with selectivity for the lizard melanocyte receptors over the frog melanocyte receptors. Increasing or decreasing the ring size of these cyclic melanotropins from 23 diminishes the biological potency of the resulting cyclic peptide. The 23- and 24 membered ring analogues showed prolonged (residual) biological activities in both biological assays, but the smaller ring systems (20, 21, 22) did not. These results provide new insights into the structural and conformational requirements of alpha-MSH and its analogues at two different types of pigment cell (melanocyte) receptors. PMID- 2555513 TI - Central nervous system malformations in Mohr's syndrome. AB - A boy with severe developmental delay, bilateral, symmetrical hallucal duplication, and accessory alveolar frenula was found to have radiological evidence of a large arachnoid cyst compressing the cerebellum and brain stem. We review neurological abnormalities in Mohr's syndrome. PMID- 2555514 TI - Contrast definition and contour detection for logarithmic images. AB - Logarithmic images, such as images obtained by transmitted light or those produced by the human visual system, differ from linear images. Their processing and analysis require consequently specific laws and structures. The latter have been developed in the concept of a logarithmic image processing (LIP) model (Jourlin & Pinoli, 1987, 1988; Pinoli, 1987a). This model permits the introduction of a well-justified contrast definition: from a physical point of view, it is closely linked with logarithmic images and from a mathematical point of view, it is set up in an algebraic structure. The applications presented at the end of this paper concern image preprocessing and segmentation. In particular, in the case of microscopic images, the proposed method of segmentation gives good results with transmitted light (thin foils in biology or transmitted electronic microscopy). However, images obtained by reflected light microscopy are not within the scope of this model. PMID- 2555515 TI - Large cell carcinoma of the lung with isolated jejunal metastasis. AB - Isolated small bowel metastasis of bronchogenic carcinoma is so distinctly unusual that only three previous case reports have been located. The authors report the case of a 59-year-old white male, who succumbed due to complications of a jejunal metastasis two months after left upper lobectomy for large cell carcinoma. The diagnosis was suspected preoperatively on the basis of a CT scan. PMID- 2555516 TI - Radiological seminar CCXLXIII: percutaneous gastrojejunostomy--a interventional radiologic procedure. AB - The authors describe the recent emergence of an interventional radiologic procedure, percutaneous gastrostomy and gastroenterostomy. They present a case report, and discuss indications, methods, and advantages of the procedure for some patients. PMID- 2555517 TI - Transposition effect of adenine (Dam) methylation on activity of O end mutants of IS50. AB - The two ends of insertion sequence IS50 (from Tn5) differ in sequence and in activity during transposition: the IS50 I end contains DNA adenine methylation (Dam) sites and is affected directly by Dam methylation, whereas the O end lacks Dam sites. The effect of Dam methylation on the transposition of IS50-derived elements with base substitution mutations in their O ends was assayed to understand better how the divergent O and I ends interact. Of 31 O end mutations tested, ten impaired transposition less, and two impaired transposition more in Dam- than in Dam+ cells. These results suggest that the interaction between the two ends in a transposition complex is affected by the sequence or the extent of methylation of one end. PMID- 2555518 TI - Structural and electronic properties of the liver fluke heme cavity by nuclear magnetic resonance and optical spectroscopy. Evidence for a distal tyrosine residue in a normally functioning hemoglobin. AB - Structural features of the heme and the heme cavity of the monomeric hemoglobin (Hb) from the platyhelminth Dicrocoelium dendriticum were investigated by optical and proton nuclear magnetic resonance spectroscopy. Using nuclear Overhauser effects (NOEs) from resonances assigned previously through isotope labeling, most hyperfine-shifted resonances could be attributed to individual heme and protein protons in the cyano-metHb complex. It was observed that the heme 2-vinyl group is held in the trans orientation by nearby residues, whereas the 4-vinyl group exhibits an equilibrium between cis and trans orientations. NOE experiments in 1H2O allowed the identification of exchangeable protons belonging to the proximal histidine residue (F8) and to a distal residue. Detailed analysis of the NOE patterns obtained from the distal labile proton to non-labile protons and among these latter protons leads to the conclusion that a tyrosine side-chain occupies the distal site E7. Optical spectra of the alkaline-metHb also lead to this view, in that they are not typical of a hydroxy-metHb complex but instead resemble that of a hemin-phenolate or human mutant (M-type) Hb with a tyrosine residue linked to the iron atom. Further evidence for a distal tyrosine residue stems from the occurrence of an unusually stable transient ferrous Hb-cyanide complex, formed upon reduction of cyano-metHb to deoxy-Hb with dithionite. We suggest that the stability of this intermediate is due to a slow re-orientation of a large distal side-chain prior to cyanide dissociation. The sequence of the E-helix, known from the partially determined primary structure, was realigned to accommodate these findings. A frame-shift by one residue now positions a tyrosine at the distal site E7 instead of the originally proposed glycine residue. PMID- 2555519 TI - Comparative sequence and immunochemical analyses of murine monoclonal anti morphine antibodies. AB - A more complete characterization is given for four previously reported anti morphine monoclonal antibodies that bind the hapten with high affinity and to which anti-idiotypic antibodies have been raised that mimic opiates at receptor binding sites. The variable (V) region nucleotide sequences of the heavy (H) and light (L) chains of these murine antibodies have been determined by direct sequencing of the poly(A)+ mRNAs using specific oligonucleotide primers and dideoxynucleotide chain-termination, and the deduced amino acid sequences are compared. The primary sequences predicted for the VH segments of 10C3 and 11C7 antibodies are closely associated with the VHIIIB subgroup of mouse H-chain (80 to 82% homology), while those for the V-regions of 3B9 and 12D4 H-chains correlate well with the VHIIC subgroup (64 to 67% homology). The 11C7, 10C3, 3B9 and 12D4 hybridoma cell lines use JH1, JH2, JH3 and JH4 DNA segments, respectively. Since considerable variations in length and primary sequence in the CDR3 (complementarity determining region) peptides of all the H-chains are evident, conservation of the D-region structure does not appear to be necessary for effective hapten binding. However, sequence homologies of the CDR2 regions of all the antibodies indicate that residues Glu H-50, Ile H-51, Pro H-52a and Tyr H 59 are conserved, and that these segments may be more critically involved in binding than the other H and L-chain hypervariable regions. The marked VL sequence homology, greater than 93%, among the L-chains and consensus lambda sequence, suggests derivation from a similar or identical VL germ-line gene. The L-chain J-region peptides for all the antibodies are classified JL1 and no VL-JL junctional diversity was apparent. The antimorphine antibody L-chains are apparently generated by the joining of a specific J-gene segment to a single germ line V-gene segment, and minor sequence variations are the result of somatic mutations within the coding region. The leader sequence for one of the H-chains was determined. The inhibition of morphine binding by phenoxybenzylation or iodination of the affinity-purified immunoglobulins indicates the involvement of a single tyrosyl residue within or close to the antibody-combining site for the opiate. This conclusion is supported by the sequence data and nuclear magnetic resonance measurements reported in the accompanying paper, in which the results are used to interpret nuclear magnetic resonance measurements on one of the ligand-antibody systems. The possible importance of additional contact amino acids, tryptophan, aspartic and/or glutamic acids, is discussed. PMID- 2555520 TI - Isolation and characterization of ribosomal DNA variants from Sciara coprophila. AB - The ribosomal RNA multigene family in the fungus fly Sciara coprophila contains a total of only 65 to 70 repeat units. We explored the types and frequencies of variant repeats in this small multigene family by characterizing different cloned rDNA variants from Sciara. Although we did not observe any intergenic spacer length variants in Sciara, we found a variant due to the insertion of a putative mobile element (lambda Bc11), and variants containing ribosomal insertion elements. By DNA sequence analysis of rDNA/non-rDNA junctions, there are three distinct types of ribosomal insertion elements found in Sciara rDNA: two correspond to the R1 and R2 insertion elements found in other dipterans (clones lambda Bc5 and pBc1L1, respectively), and one is a novel class of ribosomal insertion elements (R3, exemplified by clone pBc6D6) which so far is unique to Sciara. Together, the several different rDNA variants make up from 12 to 20% of the rDNA in Sciara. These results are discussed in the context of evolution of the ribosomal RNA multigene family. PMID- 2555521 TI - Kinetic evidence for insertion of actin monomers between the barbed ends of actin filaments and barbed end-bound insertin, a protein purified from smooth muscle. AB - An actin polymerization-retarding protein was isolated from chicken gizzard smooth muscle. This protein copurified with vinculin on DEAE-cellulose and gel filtration columns. The polymerization-retarding protein could be separated from vinculin by hydroxylapatite chromatography. The isolated polymerization-retarding protein lost its activity within a few days, but was stable for weeks when it was not separated from vinculin. We termed the polymerization-retarding protein "insertin". Because of the instability of the isolated insertin, we investigated the effect of insertin-vinculin on actin polymerization. Insertin-vinculin retarded nucleated actin polymerization maximally fivefold. Polymerization at the pointed ends of gelsolin-capped actin filaments was not affected by insertin vinculin, suggesting that insertin-vinculin binds to the barbed ends, but not to the pointed ends, of actin filaments. Retarded polymerization was observed even if the actin monomer concentration was between the critical concentrations of the ends of treadmilling actin filaments. As at this low monomer concentration the pointed ends depolymerize, monomers appeared to be inserted at the barbed ends between the terminal subunit and barbed end-bound insertin molecules. Insertin vinculin was found not to increase the actin monomer concentration to the value of the pointed ends. These observations support the conclusion that insertin is not a barbed end-capping protein but an actin monomer-inserting protein. According to a quantitative analysis of the kinetic data, all observations could be explained by a model in which two insertin molecules were assumed to bind co operatively to the barbed ends of actin filaments. Actin monomers were found to be inserted between the barbed ends and barbed end-bound insertin molecules at a rate of about 1 x 10(6) M-1 s-1. Insertin may be an essential part of the machinery of molecules that permit treadmilling of actin filaments in living cells by insertion of actin molecules between membranes and actin filaments. PMID- 2555522 TI - Crystallization and preliminary X-ray analysis of porcine synovial collagenase. AB - Crystals of porcine synovial collagenase suitable for an X-ray structure analysis have been obtained. The crystals belong to space group I4, with unit cell dimensions a = b = 160.0 A, c = 53.1 A, with one molecule in the asymmetric unit. Diffraction extends beyond 3 A perpendicular to the c axis but along the 4-fold axis, the intensities are measurable only to 4 A. PMID- 2555524 TI - Detection of mitochondrial membrane damages in myocardial ischemia with ESR spin labeling technique. AB - The ultrastructure and membrane fluidity were examined in two fractions of myocardial mitochondria isolated from the ischemic region following occlusion of the circumflex coronary artery of the dog. Transmission electron microscopy revealed that 60 min of ischemia produced more structural change in the subsarcolemmal mitochondria than in the intermyofibrillar mitochondria. Changes in the dynamic properties of the mitochondrial membranes were examined using the spin labeling technique with electron spin resonance (ESR). From the ESR spectra, the membrane fluidity of the subsarcolemmal mitochondria was found to decrease significantly after ischemic injury, whereas no difference was observed in intermyofibrillar mitochondria. In addition, a negative correlation between the increase of order parameter S and dysfunction of respiratory responses of subsarcolemmal mitochondria was found. These results suggest that the two fractions of mitochondria in the myocardium showed different responses to acute ischemic injury. The damage to mitochondrial membranes occurred during the period of ischemia. PMID- 2555523 TI - Crystal structure of human rhinovirus serotype 1A (HRV1A). AB - The structure of human rhinovirus 1A (HRV1A) has been determined to 3.2 A resolution using phase refinement and extension by symmetry averaging starting with phases at 5 A resolution calculated from the known human rhinovirus 14 (HRV14) structure. The polypeptide backbone structures of HRV1A and HRV14 are similar, but the exposed surfaces are rather different. Differential charge distribution of amino acid residues in the "canyon", the putative receptor binding site, provides a possible explanation for the difference in minor versus major receptor group specificities, represented by HRV1A and HRV14, respectively. The hydrophobic pocket in VP1, into which antiviral compounds bind, is in an "open" conformation similar to that observed in drug-bound HRV14. Drug binding in HRV1A does not induce extensive conformational changes, in contrast to the case of HRV14. PMID- 2555525 TI - Inotropic and lusitropic effects of MCI-154 (6-[4-(4- pyridyl)aminophenyl]-4,5 dihydro-3(2H)-pyridazinone) on human myocardium. AB - We studied the inotropic and lusitropic responses to MCI-154 in 12 right or left ventricular trabeculae carneae isolated from 7 organ donors (non-cardiac) without known cardiovascular disease who met accepted criteria for brain death. Isometric tension was recorded from muscles superfused with a physiologic salt solution at 30 degrees C, and stimulated to contract at three-second intervals. Concentration response curves were developed over a range of MCI-154 organs bath concentrations (10(-7) M to 3 x 10(-4) M; n = 9). Six experiments were conducted using 10(-6) M carbachol, a muscarinic agonist, in the presence of a maximally effective concentration of MCI-154 to test for dependence of tension development on cyclic adenosine monophosphate. Three experiments were conducted with MCI-154, 3 x 10( 5) M, in muscles loaded with the bioluminescent calcium indicator aequorin. MCI 154 produced a concentration-dependent rise in peak tension in the human muscle (positive inotropic effect), equivalent to 70% of the maximal response to calcium (P less than 0.001). Relaxation was enhanced (positive lusitropic effect), as evidenced by a fall in the time to 80% relaxation from 311 +/- 13 ms (baseline) to 248 +/- 15 ms at 10(-5) M (P less than 0.01). Aequorin studies showed the increase in tension to be accompanied by large increases in cystolic calcium, the principal mechanism of action. Carbachol caused MCI-154--induced maximum peak tension to decrease by 5 +/- 1%. While not excluding a cyclic adenosine monophosphate--mediated MCI action, this modest carbachol inhibition suggests the existence of additional mechanism(s) of action. MCI-154 had a negative lusitropic effect at high concentrations (greater than 10(-4)M) which may have been due to intracellular calcium overload, evidenced by the large amplitude aequorin signals. This does not exclude sensitization of the myofilaments to calcium as a possibility. Extrapolated to the in vivo setting, these experiments suggest that MCI-154 may be an effective positive inotropic agent in man. PMID- 2555526 TI - Isoproterenol sensitivity of isolated cardiac myocytes from rats with monocrotaline-induced right-sided hypertrophy and heart failure. AB - Rats treated with the alkaloid monocrotaline developed right ventricular hypertrophy with a left:right ventricle weight ratio of 1.35 +/- 0.10 (mean +/- s.e.m., n = 25) compared with 3.83 +/- 0.40 (n = 14) in diet-matched controls (P less than 0.001). Urine volume and sodium content were reduced and body water increased consistent with heart failure. In 10 out of 26 treated rats pleural, pericardial or peritoneal effusions were present. Urine norepinephrine content was significantly raised (P less than 0.02) but epinephrine was unchanged. Plasma norepinephrine levels were raised though not significantly. Myocytes isolated from the right ventricle had a reduced myosin Ca2+-activated ATPase (P less than 0.05) activity and a shift towards slower V2 and V3 myosin isoforms. There was no decrease in maximum contraction amplitude with calcium or isoproterenol in either left or right ventricular cells of treated rats. Right ventricular cells from treated rats showed a reduced rate of contraction in maximum isoproterenol (P less than 0.05) and a significant rightward shift in PD2 (P less than 0.05) representing a two-fold increase in EC50 for isoproterenol compared with right ventricular cells from control animals. There was no shift in EC50 for isoproterenol in left ventricle cells. In parallel experiments, myocytes isolated from both ventricles of rats treated with isoproterenol for one week showed a rightward shift of more than 50-fold in the isoproterenol concentration-response curve and a depressed response to maximum isoproterenol. In the rat monocrotaline model of right-sided cardiac hypertrophy and failure, changes in sensitivity to beta-adrenoceptor agonists are slight, and present only in the right ventricle. The lack of change in the left ventricle seems to suggest that this functional desensitisation is not a consequence of raised circulating catecholamines. PMID- 2555527 TI - Effect of atrial natriuretic factor on angiotensin converting enzyme. AB - We studied the effects of atrial natriuretic factor on the conversion of angiotensin I to angiotensin II. Atrial natriuretic factor had a novel effect on angiotensin I conversion. Pulmonary artery endothelial cells converted 1.22 nmol/min/dish of [125I]angiotensin I to II in the absence of atrial natriuretic factor, but this activity was suppressed by atrial natriuretic factor. When atrial natriuretic factor was added to pulmonary artery endothelial cells, the conversion of angiotensin I to angiotensin II was suppressed 0.475 nmol/min/dish at 10(-6) M of atrial natriuretic factor. PMID- 2555529 TI - CDC moves on chronic diseases. PMID- 2555528 TI - Inotropic effect of phenylephrine and myocardial alpha-adrenergic receptor in newborn and adult animals. AB - Developmental changes in the myocardial alpha-receptor density were studied using rabbit, rat and dog hearts. In all species studied, alpha-receptor density in the newborn was greater than in the adult. The inotropic effect of phenylephrine was measured using the isolated arterially perfused heart preparation of rabbit and rat. The heart was stimulated electrically at 40/min. In the presence of propranolol, phenylephrine caused a significant positive inotropic effect which was significantly less in the newborn than in the adult. Since alpha-adrenergic stimulation activates protein kinase C, the inotropic effect of protein kinase C activation was studied in the rabbit and rat using phorbol myristate acetate (PMA). PMA caused a negative inotropic effect and the decrease in contractile function in the newborn was greater than in the adult. These data suggest that myocardial alpha-receptor density decreases and the positive inotropic effect of alpha-agonist increases with development. The reasons for this discrepancy remain unclear but there may be developmental differences in the signal transduction processes of alpha-stimulation. The greater negative inotropy of protein kinase C activation in the premature heart may be one of the mechanisms of the reduced inotropy of alpha-agonist in this age group. PMID- 2555530 TI - Altered regulation of the cytochrome P4501A1 gene: novel inducer-independent gene expression in pulmonary carcinoma cell lines. AB - The cytochrome P450 (CYP) systems catalyze the metabolic transformation of a wide variety of xenobiotics including procarcinogens present in cigarette smoke condensate as well as atmospheric pollutants. The CYP1A1 isoenzyme is of particular interest because it has been implicated as a risk factor in the etiology of lung cancer in heavy cigarette smokers. The identification and expression of the structural CYP1A1 gene in either normal human lung or lung cancer cells has not been reported. Because of its potential significance in human lung cancer, we investigated the expression of the CYP1A1 structural gene in 24 established human lung cancer cell lines including 15 non-small cell (eight adenocarcinomas, three large cell undifferentiated carcinomas, two bronchioloalveolar cell carcinomas, and two squamous cell carcinomas) and nine small cell lung carcinomas. CYP1A1 mRNA was detected in 14 of 15 (93%) of the non small cell lung carcinoma cell lines examined following 24-hour treatment with benz[a]anthracene (BA) and in nine of 15 (60%) of the non-small cell lines cultured without an inducer in the medium. When the small cell lung cancer lines were evaluated for CYP1A1 gene expression, two of nine (22%) expressed detectable CYP1A1 mRNA in both BA-induced cell cultures and constitutive (control) cultures. A positive correlation was noted between BA-induced CYP1A1 mRNA levels and the corresponding aryl hydrocarbon hydroxylase activity expressed as absolute BA induced enzyme activity (r = 0.74; P less than .01; n = 24), which further demonstrated that CYP1A1 mRNA expression reflects CYP1A1 enzyme activity in the individual cell lines. These observations represent the first known demonstration of constitutive (non-induced) CYP1A1 gene expression in human cells and suggest altered regulation of the CYP1A1 gene in selected lung cancer cell lines. These human pulmonary carcinoma cell lines, which have documented regulatory defects, could be useful for further identification of the mechanisms associated with CYP1A1 gene regulation. PMID- 2555531 TI - Smoking, air pollution, and the high rates of lung cancer in Shenyang, China. AB - A case-control study involving interviews with 1,249 patients with lung cancer and 1,345 population-based controls was conducted in Shenyang, an industrial city in northeastern China, where mortality rates are high among men and women. Cigarette smoking was found to be the principal cause of lung cancer in this population, accounting for 55% of the lung cancers in males and 37% in females. The attributable risk percentage among females is high compared to elsewhere in China, largely because of a higher prevalence of smoking among women. After adjustment for smoking, there were also significant increases in lung cancer risk associated with several measures of exposure to air pollutants. Risks were twice as high among those who reported smoky outdoor environments, and increased in proportion to years of sleeping on beds heated by coal-burning stoves (kang), and to an overall index of indoor air pollution. Threefold increases in lung cancer risk were found among men who worked in the nonferrous smelting industry, where heavy exposures to inorganic arsenic have been reported. The associations with both smoking and indoor air pollution were stronger for squamous cell and small cell carcinomas than for adenocarcinoma of the lung. Risks due to smoking or air pollution were not greatly altered by adjustment for consumption of fresh vegetables or sources of beta carotene or retinol, prior chronic lung diseases, or education level. The findings suggest that smoking and environmental pollution combine to account for the elevated rates of lung cancer mortality in Shenyang. PMID- 2555532 TI - Glycerophosphorylcholine phosphocholine phosphodiesterase activity of rat brain myelin. AB - Myelin isolated from rat brain possessed the ability to release phosphorylcholine from glycerophosphorylcholine, and this activity was enriched 3.2-fold over that of the original homogenate. This glycerophosphorylcholine phosphocholine phosphodiesterase activity had a pH optimum at 9.5, had a Km of 0.2 mM, and a Vmax of 150 nmoles/mg protein/hr. The enzyme had a specific requirement for Zn+2 with an optimum concentration at 0.25 mM. Maximum enzyme activity was at 50 degrees C and an Arrhenius plot showed a breakpoint at 40 degrees. p Nitrophenylphosphorylcholine was also hydrolyzed by purified myelin and was a competitive inhibitor of glycerophosphorylcholine phosphocholine phosphodiesterase activity with a Ki of 0.075 mM. Glycerolphosphorylethanolamine was hydrolyzed only 5% compared with GPC, but it was not an inhibitor. PMID- 2555533 TI - Cyclic adenosine monophosphate as a second messenger in horizontal cell uncoupling in the teleost retina. AB - The reduction in the receptive field of horizontal cells of the teleost Eugerres plumieri observed upon dopamine (DA) superfusion is thought to be due to cell uncoupling. The possible mechanisms by which activation of DA receptors modify the electric coupling between horizontal cells were studied in the present work. It was found that the effect of DA in different preparations is mediated by a modification of intracellular concentration of cAMP and H+. The effects of intracellular injection of cAMP and H+ were studied in retinal horizontal cells of the teleost E. plumieri. A triple microelectrode was used to inject the ion iontophoretically, to pass current pulses, and to record voltages from the same cell, while a fourth microelectrode was used to record voltages from a neighboring cell in the same retinal layer. Responses evoked by light spots and annuli were evaluated simultaneously. Coupling ratios between neighboring horizontal cells ranged from 0.22 to 0.45. The intercellular resistance (Rc), 0.5 3.5 x 10(6) ohms, and that of the remaining cell membrane resistance (Rm), 2.5-18 x 10(6) ohms, were calculated by means of a passive electrical model that has a hexagonal array. The microinjection of H+ with injection current from +5 to +30 nA for 40 to 100 sec led to temporary and reversible light response reduction. The coupling ratio between two impaled cells was reduced by about 30%, and intercellular resistance (Rc) increment was 320% while cell membrane resistance (Rm) did not change consistently. There was also a temporary and reversible Rm reduction (70-85%) and an Rc increment of 170-330% when cyclic adenosine monophosphate was iontophoretically injected with current from -30 to -40 nA for 50 to 170 sec. The coupling ratio between two impaled cells was reduced by about 40%, and light responses recorded from the injected cell showed a reduction in amplitude with the same time course as that of the resistive changes. The injection of Lucifer yellow into a horizontal cell under normal conditions always results in pronounced fluorescence for more distant cells; however, under constant injection of H+ or cAMP only the injected cell is fluorescent, which provides direct evidence of the reduction in the effectiveness of coupling between horizontal cells. The observed effects of intracellular H+ or cAMP injection correspond to the resistive changes in Rc and coupling ratio that occur in the horizontal cell network upon superfusion with a dopamine (DA) solution. PMID- 2555534 TI - Rubella and cytomegalovirus (CMV) infections: laboratory aspects of investigation of antenatal, congenital, persistent, and subclinical infections. AB - Rubella specific IgM tests carried out on pregnant women with history of rubella contact or rubella-like rash indicated the presence of rubella-IgM by the second week after contact, persistence to 3-4 weeks followed by a decline and non detectability around 8-9 weeks and at delivery. Laboratory investigation of cases of rubella infection in infants and children, including clinically proven and suspected congenital rubella revealed distinct patterns of combinations of positivity and negativity of IgM and IgG antibodies. Three cases of persistence of rubella specific IgM antibodies with one even up to 3 years in congenital rubella and a case of CMV-IgM persistence in congenital CMV are described. Rubella-IgM and CMV-IgM were detected in the serum of two patients aged 12 years and 24 years with CMV mononucleosis. Utilization of rubella-IgM/CMV-IgM tests enabled the identification of four cases of subclinical rubella and one of subclinical CMV in a pediatric population. PMID- 2555535 TI - The efficacy of ceruminolytics: everything old is new again. AB - Impacted cerumen frequently poses a problem to both patients and physicians. Over the years, many topical ceruminolytics have been advocated, many of which are currently used with no scientific proof of their effectiveness. The effects of a variety of ceruminolytics were tested in vitro on standard spheres of normal human cerumen. The only truly effective ceruminolytics had an aqueous base, and the most effective ceruminolytic was a 10% solution of sodium bicarbonate. In contrast, those ceruminolytics which had an organic base, including most of the commonly available commercial preparations, had little ceruminolytic effect. When we consider that Joseph Toynbee recommended the use of sodium bicarbonate ear drops as a ceruminolytic in 1860, then surely, when looking for the ideal ceruminolytic, "everything old is new again". PMID- 2555536 TI - Depletion of glycoprotein gp85 from virosomes made with Epstein-Barr virus proteins abolishes their ability to fuse with virus receptor-bearing cells. AB - Entry of an enveloped virus such as Epstein-Barr virus (EBV) into host cells involves fusion of the virion envelope with host cell membranes either at the surface of the cell or within endocytic vesicles. Previous work has indirectly implicated the EBV glycoprotein gp85 in this fusion process. A neutralizing monoclonal antibody to gp85, F-2-1, failed to inhibit binding of EBV to its receptor but interfered with virus fusion as measured with the self-quenching fluorophore octadecyl rhodamine B chloride (R18) (N. Miller and L. M. Hutt Fletcher, J. Virol. 62:2366-2372, 1988). To test further the hypothesis that gp85 functions as a fusion protein, EBV virion proteins including or depleted of gp85 were incorporated into lipid vesicles to form virosomes. Virosomes were labeled with R18, and those that were made with undepleted protein were shown to behave in a manner similar to that of R18-labeled virus. They bound to receptor-positive but not to receptor-negative cells and fused with Raji cells but not with receptor-positive, fusion-incompetent Molt 4 cells; monoclonal antibodies that inhibited binding or fusion of virus inhibited binding and fusion of virosomes, and virus competed with virosomes for attachment to cells. In contrast, virosomes made from virus proteins depleted of gp85 by immunoaffinity chromatography remained capable of binding to receptor-positive cells but failed to fuse. These results are compatible with the hypothesis that gp85 is actively involved in the fusion of EBV with lymphoblatoid cell lines and suggest that the ability of antibody F-2-1 to neutralize infectivity of EBV represents a direct effect on the function of gp85 as a fusion protein. PMID- 2555537 TI - Functional analysis of the Tat trans activator of human immunodeficiency virus type 2. AB - The trans-activator (Tat) proteins of the related but distinct type 1 and type 2 human immunodeficiency viruses (HIV-1 and HIV-2) display incomplete functional reciprocity. One possible explanation of this observation, suggested by computer analysis of potential RNA secondary structures within the viral trans-activation response (TAR) elements, is that HIV-2 Tat requires the presentation of two viral RNA stem-loop sequences for full activity whereas HIV-1 Tat is maximally active upon presentation of a single stem-loop structure. Here, we demonstrate that the HIV-2 long terminal repeat indeed contains two functionally independent TAR elements. However, the second (3') TAR element of HIV-2 is significantly less active than the 5' TAR element and is functionally masked in the context of an intact HIV-2 long terminal repeat. Evidence is presented suggesting that the activities of these two HIV-2 TAR elements reflect, at least in part, their relative distances from the site of transcription initiation. Although the HIV-2 TAR element proximal to the viral mRNA cap site appears to be sufficient for effective trans activation by HIV-2 Tat in vitro, this functional redundancy may nevertheless serve to enhance HIV-2 replication in infected cells in vivo. PMID- 2555539 TI - The essential 65-kilodalton DNA-binding protein of herpes simplex virus stimulates the virus-encoded DNA polymerase. AB - The 65-kilodalton DNA-binding protein (65KDBP) of herpes simplex virus type 1 (HSV-1), the product of the UL42 gene, is required for DNA replication both in vitro and in vivo, yet its actual function is unknown. By two independent methods, it was shown that the 65KDBP stimulates the activity of the HSV-1 encoded DNA polymerase (Pol). When Pol, purified from HSV-1-infected cells, was separated from the 65KDBP, much of its activity was lost. However, addition of the 65KDBP, purified from infected cells, stimulated the activity of Pol 4- to 10 fold. The ability of a monoclonal antibody to the 65KDBP to remove the Pol stimulating activity from preparations of the 65KDBP confirmed that the activity was not due to a trace contaminant. Furthermore, the 65KDBP did not stimulate the activity of other DNA polymerases derived from T4, T7, or Escherichia coli. The 65KDBP gene transcribed in vitro from cloned DNA and translated in vitro in rabbit reticulocyte lysates also was capable of stimulating the product of the pol gene when the RNAs were cotranslated. The product of a mutant 65KDBP gene missing the carboxy-terminal 28 amino acids exhibited wild-type levels of Pol stimulation, while the products of two large deletion mutants of the gene could not stimulate Pol activity. These experiments suggest that the 65KDBP may be an accessory protein for the HSV-1 Pol. PMID- 2555540 TI - Cleavage of small peptides in vitro by human rhinovirus 14 3C protease expressed in Escherichia coli. AB - The 3C region of human rhinovirus 14 was expressed in Escherichia coli. The microbially synthesized protease was functional, since the expressed precursor underwent autoproteolytic processing to generate mature molecules of the expected molecular weight and antigenicity. Mutation of the putative active-site Cys-146 residue to an alanine resulted in the synthesis of unprocessed precursor molecules. Large quantities of the 20-kilodalton protease were purified by a simple purification protocol, and the resulting molecule was shown to be biologically active in vitro against synthetic peptides corresponding to the 2C 3A cleavage site. This site was cleaved with high efficiency and fidelity and was used to generate kinetic data on the 3C protease. The protease exhibited sensitivity to Zn2+, was capable of cleaving five of seven rhinovirus cleavage site peptides tested with variable efficiency, and could distinguish authentic substrate peptides from control peptides containing the dipeptide cleavage sequence pair Gln-Gly. PMID- 2555541 TI - Challenge of chimpanzees (Pan troglodytes) immunized with human immunodeficiency virus envelope glycoprotein gp120. AB - The human immunodeficiency virus type 1 (HIV-1), the causative agent of acquired immunodeficiency syndrome, infects humans and chimpanzees. To determine the efficacy of immunization for preventing infection, chimpanzees were immunized with gp120 purified from human T-cell lymphotrophic virus type IIIB (HTLV-IIIB) infected cell membranes and challenged with the homologous virus, HTLV-IIIB. A challenge stock of HTLV-IIIB was prepared by using unconcentrated HTLV-IIIB produced in H9 cells. The titer of the virus from this stock on human and chimpanzee peripheral blood mononuclear cells and in human lymphoid cell lines was determined; a cell culture infectivity of 10(4) was assigned. All chimpanzees inoculated intravenously with 40 cell culture infectious units or more became infected, as demonstrated by virus isolation and seroconversion. One of two chimpanzees inoculated with 4 cell culture infectious units became infected. Chimpanzees immunized with gp120 formulated in alum developed antibodies which precipitated gp120 and neutralized HTLV-IIIB. Peripheral blood mononuclear cells from gp120-vaccinated and HIV-infected animals showed a significantly greater response in proliferation assays with HIV proteins than did peripheral blood mononuclear cells from nonvaccinated and non-HIV-infected chimpanzees. Two of the gp120-alum-immunized chimpanzees were challenged with virus from the HTLV-IIIB stock. One animal received 400 cell culture infectious units, and one received 40 infectious units. Both animals became infected with HIV, indicating that the immune response elicited by immunization with gp120 formulated in alum was not effective in preventing infection with HIV-1. PMID- 2555538 TI - Inducible expression of encephalomyocarditis virus 3C protease activity in stably transformed mouse cell lines. AB - An inducible expression vector system has been developed to facilitate the study of the effects of individual virus gene products on cell function. The vector utilizes the mouse metallothionein promoter carried on the bovine papillomavirus genome. Conditions which optimize the induced expression of open reading frames inserted downstream from the mouse metallothionein promoter have recently been described. In this communication we describe the use of this system to clone and express the encephalomyocarditis virus 3C protease in cultured mouse cells. Stably transformed cell lines could be induced to produce levels of 3C protease activity comparable to those observed during normal virus infection. In spite of this, no effects on cellular protein synthesis rate or membrane permeability were observed. It was also discovered that 3C protease as well as 3C protease containing polyproteins are turned over. This was true not only in the induced cell clones, but also during the normal course of encephalomyocarditis virus infection, as well as in translation systems in vitro. This phenomenon was highly specific for this family of polypeptides, perhaps explaining their apparent lack of cytotoxic effects. PMID- 2555542 TI - Novel 12-O-tetradecanoylphorbol-13-acetate-responsive elements in the upstream sequence of the MS gene promoter of Epstein-Barr virus. AB - We have demonstrated that gene expression from the promoter of the Epstein-Barr virus (EBV) MS gene with its upstream sequence is inducible by 12-O tetradecanoylphorbol-13-acetate (TPA). By transfecting mammalian cells with plasmids in which the MS promoter and its upstream sequence are linked to the bacterial chloramphenicol acetyltransferase gene, we have shown that treatment of the cells with TPA stimulates the expression of chloramphenicol acetyltransferase activity in both EBV-negative and -positive cell lines. This TPA response requires the cis-acting sequence between nucleotides 84440 and 85046 of the EBV genome, located either upstream or downstream of the MS promoter. The TPA induction is at the transcriptional level. When this sequence is linked to the promoter of the human herpesvirus 1 thymidine kinase gene, it can also enhance the expression of, and confer TPA responsiveness on, the thymidine kinase promoter. By constructing and transfecting mutants with 5' and 3' deletions, we have identified two TPA-responsive elements, one located between -726 and -690 and the other located between -603 and -546 relative to the transcription start site. These two sequences do not contain any homology to the previously defined elements for TPA response and may play an important role in EBV induction by TPA. PMID- 2555543 TI - Inhibition of translation in cells infected with a poliovirus 2Apro mutant correlates with phosphorylation of the alpha subunit of eucaryotic initiation factor 2. AB - A poliovirus type 2 Lansing mutant was constructed by inserting 6 base pairs into the 2Apro region of an infectious cDNA clone, resulting in the addition of a leucine and threonine into the polypeptide sequence. The resulting small-plaque mutant, 2A-2, had a reduced viral yield in HeLa cells and synthesized viral proteins inefficiently. Infection with the mutant did not lead to specific inhibition of host cell protein synthesis early in infection, and this defect was attributed to a failure to induce cleavage of the cap-binding complex protein p220. At late times after infection with the mutant virus, both cellular and viral protein syntheses were severely inhibited. To explain this global inhibition of protein synthesis, the phosphorylation state of the alpha subunit of eucaryotic initiation factor 2 (eIF-2 alpha) was examined. eIF-2 alpha was phosphorylated in both R2-2A-2- and wild-type-virus-infected cells, indicating that poliovirus does not encode a function that blocks phosphorylation of eIF-2 alpha. The kinetics and extent of eIF-2 alpha phosphorylation correlated with the production of double-stranded RNA in infected cells, suggesting that eIF-2 alpha is phosphorylated by P1/eIF-2 alpha kinase. When HeLa cells were infected with R2 2A-2 in the presence of 2-aminopurine, a protein kinase inhibitor, much higher virus titers were produced, cleavage of p220 occurred, and host cell protein synthesis was specifically inhibited. Since phosphorylation of eIF-2 alpha was not inhibited by 2-aminopurine, we propose that 2-aminopurine rescues the ability of R2-2A-2 to induce cleavage of p220 by inhibition of a second as yet unidentified kinase. PMID- 2555544 TI - Phosphorylation sites of the E2 transcriptional regulatory proteins of bovine papillomavirus type 1. AB - The E2 open reading frame of bovine papillomavirus type 1 (BPV-1) encodes three transcriptional regulatory proteins. The full-length open reading frame encodes a protein of 410 amino acids which functions as a transcriptional transactivator. Two transcriptional repressor proteins, E2-TR and E8/E2, contain the C-terminal 249 and 204 amino acids, respectively. We have expressed both the full-length E2 protein and the E2-TR repressor protein in insect cells, by using recombinant baculoviruses, and in mammalian COS-1 cells, by using a chimeric simian virus 40/BPV-1 virus. Analysis of the E2 proteins revealed that both the transactivator and repressor forms are phosphorylated predominately on serine residues at similar sites in both expression systems. By a combination of peptide mapping and site-directed mutagenesis techniques, the serine residues at positions 298 and 301 were determined to be the major phosphorylation sites of the BPV-1 E2 proteins. PMID- 2555545 TI - FH3, a v-myc avian retrovirus with limited transforming ability. AB - We have isolated a new acute avian transforming virus which contains the oncogene myc. This virus, designated FH3, was isolated after injection of a 10-day-old chick embryo with avian leukosis virus. While FH3 shares many properties with other v-myc-containing avian retroviruses, it also has several unique properties. The primary target for transformation in vitro is chicken macrophages; infection of chicken fibroblasts does not lead to complete morphological transformation. FH3 also exhibits a limited host range, in that Japanese quail macrophages and fibroblasts are infected but are not completely transformed. FH3 induces in vivo a limited tumor type if injected into 10-day-old chick embryos; only a cranial myelocytoma, which does not appear to be metastatic, can be detected. The v-myc gene of FH3 is expressed predominantly as a P145 Gag-Myc protein which is encoded by a ca. 8-kilobase genomic RNA. This FH3-encoded polyprotein is localized in the nucleus of all infected cells, whether or not they are transformed. PMID- 2555547 TI - Role of heterologous and homologous glycoproteins in phenotypic mixing between Sendai virus and vesicular stomatitis virus. AB - Phenotypic mixing between Sendai virus and vesicular stomatitis virus (VSV) or the mutant VSV ts045 was studied. Conditions were optimized for double infection, as shown by immunofluorescence microscopy. Virions from double-infected cells were separated by sequential velocity and isopycnic gradient centrifugations. Two types of particles with mixed protein compositions were found. One type was VSV particles with Sendai virus spikes, i.e., phenotypically mixed particles. A second type was Sendai virus-VSV associations, which in plaque assays also behaved as phenotypically mixed particles. The ratio of VSV G protein to Sendai virus glycoproteins on the cell surface was varied, using the VSV mutant ts045 in double infections. Thus, different amounts of the VSV G protein were allowed to reach the cell surface at 32, 38, and 39 degrees C in Sendai virus-infected cells. However, a fixed number of Sendai virus spikes was always found in the ts045 virions. This represented 12 to 16% of the number of G proteins present in normal VSV. Furthermore, the yield of ts045 virions was radically reduced during double infection when the temperature was raised to block G-protein transport to the cell surface, suggesting that the Sendai virus glycoproteins were not able to compensate for G protein in budding. These results emphasize the role of the G protein in VSV assembly. PMID- 2555546 TI - Mapping the termini and intron of the spliced immediate-early transcript of equine herpesvirus 1. AB - Equine herpesvirus 1 (EHV-1) has been shown to synthesize a 6.0-kilobase (kb) species of immediate-early (IE) mRNA in productively infected cells. This IE gene region maps within the outer portion (map units 0.79 to 0.83 and 0.96 to 1.00) of the two inverted repeat segments of the short genomic region, and elucidation of its DNA sequence has revealed multiple potential open reading frames (ORFs), including a major ORF of 4,461 nucleotides (F. J. Grundy, R. P. Baumann, and D. J. O'Callaghan, Virology 172:223-236, 1989). Analyses of IE polypeptides synthesized in EHV-1-infected cells (in vivo) and in vitro translation of hybrid selected IE mRNA indicated that multiple species of IE proteins are encoded by this IE mRNA species. To address the nature of the 6.0-kb IE RNA species, Northern (RNA) blot hybridization, S1 nuclease mapping, and primer extension analyses have been employed. These data revealed that no major introns were detected within the body of the IE transcript. However, the IE mRNA was shown to be spliced at the 5' terminus, such that a 372-base intron containing two small ORFs (19 and 51 amino acids) was removed from the leader region of the transcript. This splicing event reduced the leader region from 625 to 253 bases. S1 and primer extension analyses of the 5' terminus of this transcript revealed that the transcription initiation site is located 24 to 26 bases downstream of the consensus TATAAA motif. The 3' transcription termination site was mapped by S1 nuclease analysis to approximately 10 to 20 bases downstream of the polyadenylation signal, AATAAA. The distance from the stop codon of the major ORF to the polyadenylation site is approximately 300 bases. Results from S1 nuclease experiments indicated that splicing does not occur at the 3' terminus. These studies indicated that the EHV-1 6.0-kb IE mRNA is spliced at the 5' terminus and that alternative splicing of this transcript may function in regulating translation of the IE mRNA species. PMID- 2555548 TI - Topoisomerase I sites cluster asymmetrically at the ends of the simian virus 40 core origin of replication. AB - In vivo, topoisomerase I cleavage sites are located predominantly on the strands of simian virus 40 DNA that are the templates for discontinuous synthesis (S.E. Porter and J.J. Champoux, Mol. Cell. Biol. 9:541-550, 1989). This arrangement of sites suggests that topoisomerase I may associate with replication complexes in unique functional orientations at replication forks. We have mapped topoisomerase I cleavage sites in the simian virus 40 origin of replication in vitro under conditions suitable for DNA replication. Numerous sites cluster in the inverted repeat and AT-rich domains at the ends of the core origin and are arranged on the same strands that are cut most frequently in vivo. We propose that cleavage at these sites would allow bidirectional extension of the replication bubble induced by T antigen within the core origin of replication early in the initiation of DNA synthesis. A mutational analysis of the topoisomerase I sites confirms the importance of positions -4 to -1 and +1 in the consensus sequence 5'-A/T-A/G-A/T T-break-G/A-3'. Surprisingly, more distant nucleotide positions also influence topoisomerase I sites in the inverted repeat and AT-rich domains of the core origin. The effects of distant sequences could be mediated by direct interactions with topoisomerase I or by the conformation of DNA in the core origin. PMID- 2555549 TI - Influence of asparagine-linked oligosaccharides on antigenicity, processing, and cell surface expression of herpes simplex virus type 1 glycoprotein D. AB - Glycoprotein D (gD) is an envelope component of herpes simplex virus types 1 and 2. gD-1 contains three sites for the addition of N-linked carbohydrate (N-CHO), all of which are used. Three mutants were constructed by site-directed mutagenesis, each of which altered one N-CHO addition site from Asn-X-Thr/Ser to Asn-X-Ala. A fourth mutant was altered at all three sites. The mutant genes were inserted into an expression vector, and the expressed protein was analyzed in transiently transfected COS-1 cells. The mutant protein lacking N-CHO at site 1 (Asn-94) had a reduced affinity for monoclonal antibodies (MAbs) to discontinuous epitopes, suggesting that the conformation of the protein had been altered. However, the protein was processed and transported to the cell surface. The absence of N-CHO at site 2 (Asn-121) had no apparent effect on processing or transport of gD-1 but resulted in reduced binding of two MAbs previously shown to be in group VI. Binding of other MAbs to discontinuous epitopes (including other group VI MAbs) was not affected. The absence of N-CHO at site 3 (Asn-262) had no effect on processing, transport, or conformation of the gD-1 protein. The absence of N-CHO from site 1 or from all three sites resulted in the formation of high molecular-weight aggregates or complexes and a reduction in MAb binding. However, these proteins were modified by the addition of O-glycans and transported to the cell surface. We conclude that the absence of the first or all N-linked carbohydrates alters the native conformation of gD-1 but does not prevent its transport to the cell surface. PMID- 2555550 TI - Viral DNA in horses infected with equine infectious anemia virus. AB - The amount and distribution of viral DNA were established in a horse acutely infected with the Wyoming strain of equine infectious anemia virus (EIAV). The highest concentration of viral DNA were found in the liver, lymph nodes, bone marrow, and spleen. The kidney, choroid plexus, and peripheral blood leukocytes also contained viral DNA, but at a lower level. It is estimated that at day 16 postinoculation, almost all of the viral DNA was located in the tissues, with the liver alone containing about 90 times more EIAV DNA than the peripheral blood leukocytes did. Assuming a monocyte-macrophage target, each infected cell contained multiple copies of viral DNA (between 6 and 60 copies in liver Kupffer cells). At day 16 postinoculation, most of the EIAV DNA was not integrated into host DNA, but existed in both linear and circular unintegrated forms. In contrast to acute infection, viral DNA was not detectable in tissues from asymptomatic horses with circulating antibody to EIAV. PMID- 2555551 TI - Different relative expression from two murine leukemia virus long terminal repeats in unintegrated transfected DNA and in integrated retroviral vector proviruses. AB - Results of transient-expression studies have suggested a correlation between tissue-specific pathogenicity of murine leukemia viruses and the relative transcriptional activities of their long terminal repeats in various cell types. To test whether transient-expression ratios are representative of those of integrated proviruses, we developed a system for generation of retroviral transmission vectors differing only in U3. Vectors with the long terminal repeats of leukemogenic SL3-3 and nonleukemogenic Akv viruses were used for infection of a lymphoid cell line. We then compared expression in infected cells with transient expression after DNA transfection. In contrast to a high SL3-3/Akv reporter gene expression ratio in the transient assays, the ratio in stably infected populations was low. Sets of random cell clones from the two infected populations showed wide variation, with a mean value ratio identical to the population ratio but a considerably higher ratio between lowest values. We suggest that the lower expression levels, like transient expression, reflect inherent enhancer strength and that the higher levels represent chromosomal influence. The different pathogenicity, despite the moderate difference in average expression, may then relate to a different capacity for insertional oncogene activation owing to the different inherent enhancer strengths revealed by the transient-expression assays and the least active proviruses. PMID- 2555552 TI - Simian virus 40 host range/helper function mutations cause multiple defects in viral late gene expression. AB - Simian virus 40 (SV40) deletion mutants dlA2459 and dlA2475 express T antigens that lack the normal carboxy terminus. These mutants are called host range/helper function (hr/hf) mutants because they form plaques at 37 degrees C on BSC-1 and Vero monkey kidney cell lines but not on CV-1p monkey kidney cells. Wild-type SV40 can provide a helper function to permit growth of human adenoviruses in monkey kidney cells; the hr/hf mutants cannot. Progeny yields of hr/hf mutants are also cold sensitive in all cell lines tested. Patterns of viral macromolecular synthesis in three cell lines (Vero, BSC-1, and CV-1) at three temperatures (40, 37, and 32 degrees C) were examined to determine the nature of the growth defect of hr/hf mutants. Mutant viral DNA replication was similar to that of the wild type in all three cell lines, indicating that the mutations affect late events in the viral lytic cycle. In mutant-infected Vero cells, in which viral yields were highest, late mRNA levels were similar to those observed during wild-type infection. Levels of viral late mRNA from mutant-infected CV-1 and BSC-1 cells at 32 and 37 degrees C were reduced relative to those of wild type-infected cells. The steady-state level of the major viral capsid protein, VP1, in mutant-infected CV-1 cells was reduced to the same extent as was late mRNA. The synthesis of agnoprotein could not be detected in mutant-infected CV-1 cells but was readily detected in CV-1 cells infected by wild-type SV40. Primer extension analyses indicated that most late mRNAs from mutant-infected CV-1 cells utilize start sites downstream from the major wild-type cap site (nucleotide 325) and the agnoprotein initiation codon (nucleotide 335). These results indicate that deletion of the carboxyl-terminal domain of T antigen affects viral late mRNA production, both quantitatively and qualitatively. The agnoprotein is detected late in the wild-type SV40 lytic cycle and is thought to play a role in the assembly or maturation of virions. Reduced hr/hf progeny yields could result from decreased capsid protein synthesis and, in the absence of detectable levels of agnoprotein, from inefficient use of available capsid proteins. PMID- 2555553 TI - Genetic evidence for multiple nuclear functions of the herpes simplex virus ICP8 DNA-binding protein. AB - We have isolated several mutant herpes simplex viruses, specifically mutated in the infected cell protein 8 (ICP8) gene, to define the functional domains of ICP8, the major viral DNA-binding protein. To facilitate the isolation of these mutants, we first isolated a mutant virus, HD-2, with the lacZ gene fused to the ICP8 gene so that an ICP8-beta-galactosidase fusion protein was expressed. This virus formed blue plaques on ICP8-expressing cell lines in the presence of 5 bromo-4-chloro-3-indolyl-beta-D-galactopyranoside. Mutated ICP8 gene plasmids cotransfected with HD-2 DNA yielded recombinant viruses with the mutant ICP8 gene incorporated into the viral genome. These recombinants were identified by formation of white plaques. Four classes of mutants were defined: (i) some expressed ICP8 that could bind to DNA but could not localize to the cell nucleus; (ii) some expressed ICP8 that did not bind to DNA but localized to the nucleus; (iii) some expressed ICP8 that neither bound to DNA nor localized to the nucleus; and (iv) one expressed ICP8 that localized to the cell nucleus and bound to DNA in vitro, but the mutant virus did not replicate its DNA. These classes of mutants provide genetic evidence that DNA binding and nuclear localization are distinct functions of ICP8 and that ICP8 has nuclear functions other than binding to DNA. Furthermore, the portion of ICP8 needed for a nuclear function(s) distinct from DNA binding is the part of ICP8 showing sequence similarity to that of the cellular protein cyclin or proliferating cell nuclear antigen. PMID- 2555554 TI - The Epstein-Barr virus (EBV) early protein EB2 is a posttranscriptional activator expressed under the control of EBV transcription factors EB1 and R. AB - From the cloning and characterization of cDNAs, we found that the Epstein-Barr virus (EBV) open reading frame (ORF) BMLF1-BSLF2 coding for the early protein EB2 is present in several mRNAs generated by alternative splicing and expressed in the leftward direction from two promoters PM and PM1. The PM promoter controls the expression of two abundant mRNA species of 1.9 and 2 kilobases (kb), whereas the PM1 promoter controls the expression of at least three mRNAs 3.6, 4.0, and 4.4 kb long. The PM promoter probably overlaps with the PS promoter which controls the transcription of a 3.6-kb mRNA expressed in the rightward direction and containing the ORF BSRF1. Although it increases the amount of chloramphenicol acetyltransferase enzyme expressed from the chimeric pMCAT gene, EB2 is not a promiscuous trans-activator of gene expression and does not positively regulate its own expression from promoter PM. The EB2 activation is not promoter dependent but could possibly act by stabilizing mRNAs and increasing their translation. The PM promoter is, however, activated by the two EBV transcription trans-acting factors, EB1 and R, encoded by the EBV ORFs BZLF1 and BRLF1, respectively. EB1 activates the PM promoter from a consensus AP-1 binding site, and R activates the PM promoter from an enhancer. PMID- 2555555 TI - High-frequency leader sequence switching during coronavirus defective interfering RNA replication. AB - A system was developed that exploited defective interfering (DI) RNAs of coronavirus to study the role of free leader RNA in RNA replication. A cDNA copy of mouse hepatitis virus DI RNA was placed downstream of the T7 RNA polymerase promoter to generate DI RNAs capable of extremely efficient replication in the presence of a helper virus. We demonstrated that, in the DI RNA-transfected cells, the leader sequence of these DI RNAs was switched to that of the helper virus during one round of replication. This high-frequency leader sequence exchange was not observed if a nine-nucleotide stretch of sequence (UUUAUAAAC) at the junction between the leader and the remaining DI sequence was deleted. This observation suggests that a free leader RNA generated from the genomic RNA of mouse hepatitis virus may participate in the replication of DI RNA. PMID- 2555556 TI - The avian retroviral integration protein cleaves the terminal sequences of linear viral DNA at the in vivo sites of integration. AB - The purified integration protein (IN) of avian myeloblastosis virus is shown to nick double-stranded oligodeoxynucleotide substrates that mimic the ends of the linear form of viral DNA. In the presence of Mg2+, nicks are created 2 nucleotides from the 3' OH ends of both the U5 plus strand and the U3 minus strand. Similar cleavage is observed in the presence of Mn2+ but only when the extent of the reaction is limited. Neither the complementary strands nor sequences representing the termini of human immunodeficiency virus type 1 DNA were cleaved at analogous positions. Analysis of a series of substrates containing U5 base substitutions has defined the sequence requirements for site selective nicking; nucleotides near the cleavage site are most critical for activity. The minimum substrate size required to demonstrate significant activity corresponds to the nearly perfect 15-base terminal inverted repeat. This in vitro activity of IN thus produces viral DNA ends that are joined to host DNA in vivo and corresponds to an expected early step in the integrative recombination reaction. These results provide the first enzymatic support using purified retroviral proteins for a linear DNA precursor to the integrated provirus. PMID- 2555557 TI - Oligomerization of glycolipid-anchored and soluble forms of the vesicular stomatitis virus glycoprotein. AB - The vesicular stomatitis virus glycoprotein forms noncovalently linked trimers in the endoplasmic reticulum before being transported to the Golgi apparatus. The experiments reported here were designed to determine if the extracellular domain of the glycoprotein contains structural information sufficient to direct trimer formation. To accomplish this, we generated a construct encoding G protein with the normal transmembrane and anchor sequences replaced with the sequence encoding 53 C-terminal amino acids from the Thy-1.1 glycoprotein. We show here that these sequences were able to specify glycolipid addition to the truncated G protein, probably after cleavage of 31 amino acids derived from Thy-1.1. The glycolipid anchored G protein formed trimers and was expressed on the cell surface in a form that could be cleaved by phosphoinositol-specific phospholipase C. However, the rate of transport was reduced, compared with that of wild-type G protein. A second form of the G protein was generated by deletion of only the transmembrane and cytoplasmic domains. This mutant protein also formed trimers with relatively high efficiency and was secreted slowly from cells. PMID- 2555558 TI - Sequences in the human cytomegalovirus 2.7-kilobase RNA promoter which mediate its regulation as an early gene. AB - We have studied the regulation of expression of a major human cytomegalovirus (HCMV) early transcription unit located within the long repeat of the strain AD169 genome. This region specified a 2.7-kilobase RNA which underwent its largest increase in abundance between 8 and 14 h postinfection. To study the regulation of this gene, its promoter was cloned 5' of the gene for chloramphenicol acetyltransferase (CAT) for use in transient expression assays. A construct containing 651 base pairs of upstream sequence and 54 base pairs of leader sequence was transfected into human fibroblast cells, followed by HCMV infection. Analysis of the steady-state levels of RNA expressed from this hybrid gene indicated that it accumulated with the same kinetics as the authentic viral transcript early in the infection. Cotransfection of human fibroblasts with the 2.7-kilobase RNA promoter-CAT construct and plasmids containing different HCMV immediate-early (IE) genes showed that the region of the HCMV genome encoding the transcription units corresponding to IE1 and 2 and the 5' end of IE3 is capable of stimulating promoter activity but not to the same extent as HCMV infection. To define important cis-acting regulatory elements in the promoter, a series of 5' deletion mutants was constructed. Transient expression analysis showed a stepwise reduction in inducible CAT activity, suggesting the presence of multiple regulatory sites. To further characterize the nature of these sites, we used gel mobility shift assays to study DNA-protein interactions occurring within this promoter sequence. With nuclear extracts prepared from HeLa cells as well as from infected and uninfected human foreskin fibroblasts, we found specific binding of a cellular factor to a region of the promoter important in HCMV inducible activity. This region contains a palindromic octamer with homology to the binding site of the cellular factor USF/MLTF. PMID- 2555559 TI - Polyprotein processing of Theiler's murine encephalomyelitis virus. AB - To investigate polyprotein processing of Theiler's murine encephalomyelitis viruses, we analyzed in vitro translation reactions programmed by in vitro derived transcripts from an infectious full-length cDNA clone of the DA strain of Theiler's virus. To help identify the proteinases that carried out the processing, we modified the DA cDNA clone transcription template by linearization with different restriction endonucleases that generate templates of different lengths or by constructing linker insertion or deletion mutations or both in putative proteinase-coding regions. Protein 3C carried out most of the cleavages of the polyprotein, as is true for the other picornaviruses that have been studied. A second proteinase also appeared active at the LP12A-2B junction. A protein of slightly faster mobility than the leader protein was seen with translation of transcripts derived from DA cDNA but not GDVII cDNA. This protein may be synthesized from an alternative initiation site in the DA leader-coding region out of phase with the polyprotein reading frame. Our findings are relevant to ongoing investigations of the abnormal virus expression seen in DA virus late demyelinating disease, since polyprotein processing is critical in regulating picornaviral gene expression. PMID- 2555561 TI - Attenuation and cell culture adaptation of hepatitis A virus (HAV): a genetic analysis with HAV cDNA. AB - RNA transcripts of hepatitis A virus (HAV) HM-175 cDNA from attenuated, cell culture-adapted HAV were infectious in cell culture. A full-length HAV cDNA from wild-type HAV (propagated in marmosets in vivo) was constructed. Chimeric cDNAs that contained portions of both wild-type and attenuated genomes were produced. Oligonucleotide-directed mutagenesis was used to engineer a point mutation into the VP1 gene of attenuated HAV cDNA, so that the sequence of this capsid protein would be identical to that of the wild-type virus. Transfection of monkey kidney cells with RNA transcripts from several of the chimeric cDNAs and from the mutagenized cDNA induced production of HAV. Comparison of the growth of attenuated, wild-type, chimeric, and mutant viruses in vitro indicated that the P2-P3 (nonstructural protein) region is important for cell culture adaptation of the virus; the 5' noncoding region may also contribute to adaptation, but to a lesser extent. Inoculation of marmosets with transfection-derived virus also suggested that the P2-P3 region plays an important role in attenuation of HAV HM 175. PMID- 2555560 TI - Construction of less neurovirulent polioviruses by introducing deletions into the 5' noncoding sequence of the genome. AB - Viral attenuation may be due to lowered efficiency of certain steps essential for viral multiplication. For the construction of less neurovirulent strains of poliovirus in vitro, we introduced deletions into the 5' noncoding sequence (742 nucleotides long) of the genomes of the Mahoney and Sabin 1 strains of poliovirus type 1 by using infectious cDNA clones of the virus strains. Plaque sizes shown by deletion mutants were used as a marker for rate of viral proliferation. Deletion mutants of both the strains thus constructed lacked a genome region of nucleotide positions 564 to 726. The sizes of plaques displayed by these deletion mutants were smaller than those by the respective parental viruses, although a phenotype referring to reproductive capacity at different temperatures (rct) of viruses was not affected by introduction of the deletion. Monkey neurovirulence tests were performed on the deletion mutants. The results clearly indicated that the deletion mutants had much less neurovirulence than with the corresponding parent viruses. Production of infectious particles and virus-specific protein synthesis in cells infected with the deletion mutants started later than in those infected with the parental viruses. The rate at which cytopathic effect progressed was also slower in cells infected with the mutants. Phenotypic stability of the deletion mutant for small-plaque phenotype and temperature sensitivity was investigated after passaging the mutant at an elevated temperature of 37.5 degrees C. Our data strongly suggested that the less neurovirulent phenotype introduced by the deletion is very stable during passaging of the virus. PMID- 2555562 TI - Requirements for species-specific papovavirus DNA replication. AB - Replication of papovavirus DNA requires a functional replication origin, a virus encoded protein, large T antigen, and species-specific permissive factors. How these components interact to initiate and sustain viral DNA replication is not known. Toward that end, we have attempted to identify the viral target(s) of permissive factors. The functionally defined replication origins of polyomavirus and simian virus 40, two papovaviruses that replicate in different species (mice and monkeys, respectively), are composed of two functionally distinct domains: a core domain and an auxiliary domain. The origin cores of the two viruses are remarkably similar in primary structure and have common binding sites for large T antigen. By contrast, their auxiliary domains share few sequences and serve as binding sites for cellular proteins. It seemed plausible, therefore, that if cellular permissive factors interacted with the replication origin, their targets were likely to be in the auxiliary domain. To test this hypothesis we constructed hybrid origins for DNA replication that were composed of the auxiliary domain of one virus and the origin core of the other and assessed their capacity to replicate in a number of mouse and monkey cell lines, which express the large T antigen of one or the other virus. The results of this analysis showed that the auxiliary domains of the viral replication origins could substitute for one another in DNA replication, provided that the viral origin core and its cognate large T antigen were present in a permissive cellular milieu. Surprisingly, the large T antigens of the viruses could not substitute for one another, regardless of the species of origin of the host cell, even though the two large T antigens bind to the same sequence motif in vitro. These results suggest that species specific permissive factors do not interact with the origin-auxiliary domains but, rather, with either the origin core or the large T antigen or with both components to effect DNA replication. PMID- 2555563 TI - In vitro construction of poliovirus defective interfering particles. AB - To construct poliovirus defective interfering (DI) particles in vitro, we synthesized an RNA from a cloned poliovirus cDNA, pSM1(T7)1, which carried a deletion in the genome region corresponding to nucleotide positions 1663 to 2478 encoding viral capsid proteins, by using bacteriophage T7 RNA polymerase. The RNA was designed to retain the correct reading frame in nucleotide sequence downstream of the deletion. HeLa S3 monolayer cells were transfected with the deletion RNA and then superinfected with standard virus as a helper. The DI RNA was observed in the infected cells after three passages at high multiplicity of infection. The sequence analysis of RNA extracted from the purified DI particle clearly showed that this DI RNA had the same deletion in size and location as that in the RNA used for the transfection. Thus, we succeeded in construction of a poliovirus DI particle in vitro. To gain insight into the mechanism for DI generation, we constructed poliovirus cDNAs pSM1(T7)1a and pSM1(T7)1b that, in addition to the same deletion as that in pSM1(T7)1, had insertion sequences of 4 bases and 12 bases, respectively, at the corresponding nucleotide position, 2978. The RNA transcribed from pSM1(T7)1a was not a template for synthesis of poliovirus nonstructural proteins and therefore was inactive as an RNA replicon. On the other hand, the RNA from pSM1(T7)1b replicated properly in the transfected cells. Superinfection of the transfected cells with standard virus resulted in production of DI particles derived from pSM1(T7)1b and not from pSM1(T7)1a. These observations indicate that deletion RNAs that are inactive replicons have little or no possibility of being genomes of DI particles suggesting the existence of a nonstructural protein(s) that has an inclination to function as a cis-acting protein(s). The method described here will provide a useful technique to investigate genetic information essential for poliovirus replication. PMID- 2555564 TI - Priming for rotavirus neutralizing antibodies by a VP4 protein-derived synthetic peptide. AB - In the rotavirus SA11 surface protein VP4, the trypsin cleavage sites associated with the enhancement of infectivity are flanked by two amino acid regions that are highly conserved among different rotaviruses. We have tested the ability of synthetic peptides that mimic these two regions to induce and prime for a rotavirus neutralizing antibody response in mice. After the peptide immunization schedule, both peptides induced peptide antibodies, but neither was able to induce virus antibodies, as measured by an enzyme-linked immunosorbent assay or a neutralization assay. However, when the peptide-inoculated mice were subsequently injected with intact SA11 virus, a rapid and high neutralizing antibody response was observed in mice that had previously received the peptide comprising amino acids 220 to 233 of the VP4 protein. This neutralizing activity was serotype specific; however, this peptide was also able to efficiently prime the immune system of mice for a neutralizing antibody response to the heterotypic rotavirus ST3 when the ST3 virus was used for the secondary inoculation. PMID- 2555565 TI - Fv-4 resistance gene: a truncated endogenous murine leukemia virus with ecotropic interference properties. AB - Fv-4 is a mouse gene which controls susceptibility to infection by ecotropic murine leukemia virus (MuLV). We previously cloned part of an endogenous MuLV associated with the resistance allele of the Fv-4 gene (Fv-4r). In this report, we describe an extended clone of the Fv-4r allele consisting of a 17-kilobase DNA fragment containing the retroviral sequence and its 5'-flanking sequence. The new DNA clone contains a truncated MuLV with delta pol-env-long terminal repeat sequences but no other MuLV-reactive sequence within 13 kilobases upstream of the truncated MuLV. Transfection of this clone into mouse cells led to transcription of Fv-4 env mRNA, expression of the Fv-4r-specific MuLV envelope protein, and resistance to infection with ecotropic MuLV but not amphotropic and dualtropic MuLVs. Restriction of ecotropic viruses appears to occur at or before viral cDNA synthesis. This result is consistent with a model of receptor interference for Fv 4 restriction. Our data also suggest that the 5' non-MuLV sequence is important for biological function, since a DNA clone which lacks most of the 5'-flanking sequence did not efficiently confer the resistance phenotype. PMID- 2555566 TI - The large tumor antigen of simian virus 40 encodes at least two distinct transforming functions. AB - The large tumor antigen (T antigen) of simian virus 40 is necessary and sufficient for the neoplastic transformation of a number of established cell lines. Mutational analysis has revealed that a biochemical activity residing within the amino-terminal 121 amino acids of T antigen is sufficient to induce the transformation of some cell lines, such as C3H10T1/2. The same domain of the molecule also encodes the transactivation function of T antigen and the ability to complex with the retinoblastoma susceptibility gene product. However, the transformation of other lines, such as REF52, requires an additional activity that is affected by mutations in other portions of the molecule. PMID- 2555567 TI - Release of pseudorabies virus from infected cells is controlled by several viral functions and is modulated by cellular components. AB - The role of the nonessential glycoproteins gI, gp63, and gIII in the release of pseudorabies virus from different cell lines was investigated. We show that these glycoproteins may have a beneficial or deleterious effect on virus release depending on the type of cell in which the virus is grown. Inactivation of the genes encoding either gI, gp63, or gIII has no detectable effect on virus release from rabbit kidney cells. Inactivation of gI or gp63 strongly promotes virus release from chicken embryo fibroblasts, whereas inactivation of gIII reduces virus release from these cells. A defect in both gI and gIII or in both gp63 and gIII diminishes virus release from rabbit kidney cells but improves release from chicken embryo fibroblasts. We demonstrate that all three nonessential glycoproteins contribute to one specific aspect of viral growth, namely, virus release, and that they affect virus release in conjunction with each other. Furthermore, our results show that the manifestation of the role of each of these viral functions in virus growth may differ in different cell types, i.e., that release is affected by these viral functions in conjunction with some unknown cellular function. PMID- 2555568 TI - Constitutive expression of simian virus 40 large T antigen in monkey cells activates their capacity to support polyomavirus replication. AB - Polyomavirus DNA replication is normally restricted to rodent cells, and simian virus 40 (SV40) DNA replication is restricted to primate cells. We demonstrate that DNAs containing the polyomavirus origin can be replicated in monkey cells which constitutively express SV40 large T antigen. Permissivity is most likely caused by SV40 T antigen modification of cellular protein(s) required to replicate the polyomavirus origin. A possible target for the T-antigen-induced modification is DNA polymerase alpha-DNA primase. PMID- 2555569 TI - Infectious cDNA clones of the DA strain of Theiler's murine encephalomyelitis virus. AB - The DA strain and other members of the TO subgroup of Theiler's murine encephalomyelitis viruses cause a persistent demyelinating infection, whereas the GDVII strain and other GDVII subgroup strains cause an acute lethal polioencephalomyelitis. We generated an infectious DA cDNA clone inserted into a transcription vector. Virus derived from transfection of transcripts produced a demyelinating disease indistinguishable from that of wild-type virus. The infectious clone provides a critical reagent for the production of interstrain recombinant viruses to help identify genetic loci responsible for the biological activities of the strains. PMID- 2555570 TI - Analysis of the Epstein-Barr virus origin of plasmid replication (oriP) reveals an area of nucleosome sparing that spans the 3' dyad. AB - The Epstein-Barr virus oriP, when examined with the use of in vitro nucleosome reconstitution, was found to exhibit a partial sparing of nucleosome assembly within the 3' dyad. Analysis of the homologous region containing the origin of replication of herpesvirus papio disclosed that sparing of nucleosome assembly was conserved between these two related viruses. PMID- 2555571 TI - Identification of a domain of the herpes simplex virus trans-activator Vmw65 required for protein-DNA complex formation through the use of protein A fusion proteins. AB - In order to identify structural domains of the herpes simplex virus trans activator Vmw65 required for protein-DNA complex formation, subfragments of Vmw65 were expressed in Escherichia coli as fusion polypeptides with protein A of Staphylococcus aureus, and the purified hybrids were used in a band shift assay. The results indicate that a region near the amino terminus of Vmw65 between amino acids 141 and 185 is necessary for complex formation. PMID- 2555572 TI - Vena caval resection in bulky metastatic germ cell tumors. AB - We describe the operative management and followup of vena caval resection in 12 patients with thrombosis of the inferior vena cava secondary to bulky metastatic (stage B3 or IIC) germ cell tumors. All patients received induction chemotherapy (10 platinum-based) followed by retroperitoneal lymph node dissection. The inferior vena cava was resected from just below the renal veins to the bifurcation of the iliac veins. Complete resection of retroperitoneal disease was accomplished in all patients. Five patients had postoperative complications, including 2 small bowel obstructions, 1 prolonged ileus and 2 persistent lymphatic leaks. Mean hospital stay was 37 days (range 27 to 49 days) versus 12 days (range 8 to 16 days) for noncomplicated recoveries. No long-term sequelae related to the vena caval resection have occurred with followup of 24 to 80 months. Seven patients are without disease, with a mean followup of 36 months (range 24 to 60 months). We conclude that en bloc vena caval resection for thrombosis of the vena cava allows for complete resection and simplifies the procedure with acceptable morbidity. PMID- 2555573 TI - Benign fibrous histiocytoma of the kidney. AB - A case of benign fibrous histiocytoma of the kidney in a 48-year-old man is reported. A 7-year followup has revealed no recurrence. To our knowledge there have been no previous reports of benign fibrous histiocytoma of the kidney. PMID- 2555574 TI - Experimental obstructive hydronephrosis in newborn rats. XI. A one-year follow-up study of renal function and morphology. AB - Partial obstruction of the left ureter was created in two-day-old rats and its effects on kidney function were studied with 99mTc-DMSA and 99mTc-DTPA after one, two, three and six weeks, and after one year. Kidneys from animals sacrificed at the age of six weeks or one year were also examined histologically. The obstructed renal pelvis was enlarged by about 35 times and there was a delayed excretion of 99mTc-DTPA during forced diuresis, indicating significant, chronic obstruction. The renal DMSA-uptake ratio (left kidney/(left and right kidney] was reduced to about 40% from the first week of obstruction. The parenchymal weight ratio (expressed as above) was reduced to about 45% after both six weeks and one year. The glomerular filtration rate, examined during forced diuresis and calculated on the basis of uptake capacity, was lowered to 42% after six weeks but was not significantly reduced after one year of obstruction. The incidence figures for medullary hemorrhage or accumulation of iron pigment, and chronic inflammatory changes in the cortex were somewhat higher after one year of obstruction than after 6 weeks, but the lesions were patchy in both groups. We conclude that partial unilateral ureteric obstruction, created in the neonatal period, leads to a slight but permanent functional disturbance and parenchymal weight reduction without prominent structural parenchymal damage. PMID- 2555575 TI - Government issues guidelines to stem rising tuberculosis rates in prisons. PMID- 2555576 TI - Cytomegalovirus retinitis. PMID- 2555577 TI - Human papilloma virus type 16 periungual carcinoma. PMID- 2555578 TI - Clinical and biological differences between recurrent herpes simplex virus and varicella-zoster virus infections. PMID- 2555579 TI - CMV in pregnancy case report and review of literature. PMID- 2555580 TI - [Kappa-type opioid receptor in human placental membrane]. AB - Since many opioid receptor preparations are heterogeneous systems containing multiple types of receptor, characterization of each type of the receptor is influenced by contamination with other types of receptor. Demonstrating kappa receptor proved more difficult owing to the ability of kappa ligands to interact with a number of receptor classes and to the lack of the homogeneous preparation. It has been reported that kappa ligands selectively bind to human placental membrane. To establish homogeneous kappa receptor preparation, the kappa binding to human placenta was characterized. The portion which was predominantly villus tissue was removed from freshly collected placenta, and the P3-membrane fraction was prepared. Kappa opioid agonist, such as 3H-Dynorphin A, 3H ethylketocyclazocine and 3H-U-69593, bound to the human placental membrane fraction in a manner of single class of binding site (Bmax: approximately 40 fmol.mg-1 protein). In contrast, 3H-dihydromorphine, 3H-DAGO (mu agonist), 3H DADLE (delta agonist) or 3H-SKF-10047 (sigma agonist) showed no binding activity. The specific binding of 3H-diprenorphine was displaced only by kappa-selective ligand (U-50488H, dynorphin, butorphanol). These results suggest that opioid binding in human placental membrane is specific for kappa opioid. PMID- 2555581 TI - [Effect of dexamethasone and ACTH on plasma atrial natriuretic polypeptide levels during hemorrhage in rats]. AB - Anti-shock effect of dexamethasone (Dexa) and ACTH on atrial natriuretic polypeptide (ir-ANP) secretion was studied during hemorrhage in rats as reflected in the plasma ir-ANP, corticosterone (Comp B) and ACTH levels. We also observed the number of survived rats and plasma ir-ANP levels stimulated by volume loading of the shed blood or fluid. The rats were divided into non-treated group (saline), Dexa group (Dexa 5mg.kg-1) and ACTH group (ACTH 200 micrograms.kg-1). They were anesthetized by intravenous injection of pentobarbital sodium 30 mg.kg 1. Atrial blood sample for control value was collected before hemorrhage. Approximately 20% and then 30% of the estimated blood volume was taken for hemorrhage. Hemorrhage induced significant reductions in plasma ir-ANP levels. They decreased from a control value of 133.8 +/- 6.1 pg.ml-1 to 76.6 +/- 4.2 pg.ml-1 (P less than 0.005), but plasma levels of Comp B and ACTH increased markedly. The ir-ANP secretion increased in response to the elevation of atrial pressures after volume loading. The administration of Dexa and ACTH to the rats tended to inhibit the reduction of plasma ir-ANP levels during hemorrhage and improved the survival rate 24hr after bleeding 30% of the estimated blood volume. PMID- 2555582 TI - [Natural products affecting Ca channels]. PMID- 2555583 TI - [Variety of Ca2+ channel in plasma membrane]. PMID- 2555584 TI - [Voltage-operated Ca2+ channel--its molecular structure and Ca2+ permeation]. PMID- 2555585 TI - [Current topics in receptor-dependent Ca2+ channel]. PMID- 2555586 TI - [Molecular biological studies on Ca2+ channels]. PMID- 2555587 TI - [Calcium antagonists: concepts and recent progresses in research on the mechanism of action]. PMID- 2555588 TI - [Radioreceptor binding assay of Ca antagonist]. PMID- 2555589 TI - [Effects of calcium antagonists on cardiac cells]. PMID- 2555590 TI - [Effects of Ca antagonists on Ca2+ movement in nervous system]. PMID- 2555591 TI - [ACTH secretion in adrenocortical disorders]. AB - Secretion of cortisol is under stimulatory regulation by ACTH, and cortisol secreted, in turn, inhibits ACTH secretion by the pituitary. Therefore, measurement of plasma ACTH is indispensable in the diagnosis of the adrenocortical diseases. The adrenal cortex is included in the hypothalamo adenohypophysial-adrenocortical system, and the pathogenesis of these disorders must be evaluated. An interaction between the hypothalamo-adenohypophysial adrenal system and the immune system have been suggested. We studied the effect of interleukin-1 (IL-1) on ACTH secretion by cultured rat pituitary cells in vitro. Our results suggest that IL-1 stimulates the ACTH secretion by enhancing its synthesis by the pituitary, rather than ACTH release. PMID- 2555592 TI - [Study of early gastric carcinoma from the standpoint of size]. AB - 1,112 cases of resected early gastric carcinoma without invasion beneath the propria muscularis were investigated for the location, gross findings and histological findings mainly from a standpoint of size. The incidence of intramucosal carcinomas was 92.7% in the size within 1 cm in diameter, 68.9% in 1 to 2.9 cm, 48.8% in 3 to 4.9 cm and 37.6% beyond 5 cm. Although the incidence of poorly-differentiated adenocarcinomas was 37.9% in the cases within 5 cm, the incidence in the cases beyond 5 cm was 50.8% (most of them consisted of signet ring cell carcinoma). The larger the tumor size, the more the intestinal metaplastic type carcinomas decreased and the signet-ring cell carcinomas increased. It was suggested that the signet-ring cell carcinoma infiltrates in the propria mucosae for a long time compared with the other type carcinomas. PMID- 2555594 TI - [Percutaneous transhepatic portography in patients with hepatocellular carcinoma: percutaneous transhepatic portal venous embolization]. AB - Percutaneous transhepatic portography was performed in 28 patients with HCC and PTPE was attempted in 9 of them. Displacement of the portal vein was found in 37% of 52 HCCs. Other portographic findings were: encasement (19%), avascular area (19%), occlusion (10%), narrowing (4%), neovascularization (4%), tumor staining (2%). Frequency of the tumor-related portography findings was found to be increased in relation to tumor size. TAE followed by PTPE with stainless steel coil (6 cases) or microcapsule form of MMC (3 cases) was performed on 9 cases. PTPE appears to be a promising adjuvant to conventional TAE. PMID- 2555593 TI - [Ultrasonic detectability of multiple small hepatic nodules]. AB - We analyzed the ultrasonic detectability of multiple daughter nodules accompanied by 18 hepatocellular carcinomas and multiple small nodules in 11 metastatic liver tumors identified by infusion hepatic angiography. In the cases of hepatocellular carcinoma, ultrasonic detectability of daughter nodules was remarkably limited. If any daughter nodules are detected by US, there is the great possibility that many other daughter nodules are undetected. However, US is effective in detecting small nodules in the metastatic liver tumor. PMID- 2555595 TI - [Usefulness of 99mTc-PMT cholescintigraphy in diagnosis a case of bone metastasis in hepatocellular carcinoma]. PMID- 2555596 TI - [Three cases of hepatocellular carcinoma with remarkable fatty metamorphosis]. AB - Three cases of hepatocellular carcinoma with remarkable fatty metamorphosis were reported. When we encounter lipomatous tumor with liver cirrhosis on CT and MRI, further examination is necessary because of its high possibility of hepatocellular carcinoma. PMID- 2555598 TI - [Usefulness of 5F superflexible catheter for hepatic arterial embolization]. AB - A case of hepatocellular carcinoma with marked stenosis of the celiac axis is treated by transcatheteric arterial embolization with 5F superflexible catheter. This catheter is made of polyurethane, and suitable for superselective angiography and embolization. PMID- 2555597 TI - [A case of hepatocellular carcinoma (HCC) with a huge tumor thrombus in the right atrium]. AB - A case of HCC with a huge tumor thrombus in the right atrium is reported. CT and celiac angiography revealed HCC in the liver. Inferior vena cavography and cardiac blood pool scintigraphy demonstrated a large defect in the right atrium, and 67Ga-citrate scintigraphy showed a lesion of abnormal accumulation in the right atrium. With these findings, diagnosis of HCC with a tumor thrombus in the right atrium was made, and the clinical diagnosis was confirmed by autopsy. PMID- 2555599 TI - [A case of mandibular metastasis from primary hepatocellular carcinoma]. AB - A mandibular metastasis which appeared as the first symptom from a hepatocellular carcinoma is reported. Bone metastasis related to hepatocellular carcinoma is relatively infrequent. Even less frequent is the occurrence of a mandibular metastasis as a prior symptom of hepatocellular carcinoma. In this case, also, the metastatic lesion was detected as hypervascular mass on a CT scan, thus the nature of lesion was disclosed before a biopsy was performed. PMID- 2555600 TI - [Hepatocellular carcinoma with intraductal tumor thrombus diagnosed by ultrasonography; report of two resected cases]. AB - The authors report two resected cases of hepatocellular carcinoma with intraductal tumor thrombus which showed transient jaundice and were diagnosed by ultrasonography. Ultrasonography is very useful in revealing intraductal tumor thrombus and it is necessary when patients with hepatocellular carcinoma are accompanied by jaundice. PMID- 2555601 TI - [Ultrasonic evaluation of two cases of islet cell tumor of the pancreas]. AB - Small pancreatic tumor is difficult to be diagnosed. Two cases of pancreatic islet cell tumors were examined by ultrasonography. One case was islet cell carcinoma, the other insulinoma. In both cases, ultrasonography was useful in detecting the tumor, judging the location of tumor during surgical operation and deciding the surgical procedure. PMID- 2555602 TI - [A case report of hepatoma with a giant splenic aneurysm both treated by TAE therapy]. AB - A case of a 49-year-old woman who was found to have a giant splenic aneurysm during the course of TAE therapy for hepatoma and who was given TAE therapy using 47 steel coils is reported. The splenic arterial embolization therapy using steel coils proved effective for preventing the rupture of an aneurysm and for inhibiting hypersplenism. In addition, no serious adverse effect was observed. PMID- 2555603 TI - [Typical and atypical cases of myrmecia--the differences in the nature of the inclusion bodies and the associated human papillomavirus types]. AB - A case of typical myrmecia and a case of clinically atypical, cutaneous horn-like myrmecia were studied histologically and molecular biologically. The former had eosinophilic, granular shaped inclusion bodies within the cytoplasm, as seen in other typical reported cases. In contrast, the latter was found to have more homogeneous, palely stained inclusion bodies. The analysis of human papillomavirus (HPV) DNAs detected in them clarified the association of HPV type 1 with the former and a different HPV type in the latter. These results confirmed that not only HPV type 1 but also other HPV types could produce myrmecia (inclusion wart) and suggested that the atypical case reported here might be better called by another name than myrmecia. PMID- 2555604 TI - [Leukotriene (LT) B4 and LTC4 in the blister fluid of bullous pemphigoid]. AB - Leukotriene (LT) B4 and LTC4 content in the blister fluid of patients with bullous pemphigoid (BP) was determined by radioimmunoassay. In samples from BP patients, the former (3.80 +/- 1.99 ng/ml, n = 7) significantly exceeded the amount noted in second degree burn patients (0.93 +/- 0.82 ng/ml, n = 5) and that in the fluid of suction blisters produced on normal human skin (0.95 +/- 0.21 ng/ml, n = 2). The immunoreactive LTC4 (i-LTC4) content in the blister fluid of BP (3.63 +/- 1.13 ng/ml, n = 6) was not statistically significantly different from that observed in second degree burns (2.09 +/- 1.66, n = 5), but more than that in suction blisters on normal skin (0.86 +/- 0.20, n = 2). The chemotactic activity of BP blister fluid toward polymorphonuclear leukocyte (PMNL) of BP blister fluid was determined by injecting them into guinea pig skin and observing the time course of the increase in the number of infiltrated PMNL. Neutrophils and eosinophils had accumulated in the dermis at 3 hours and at 6 hours; the numbers of each had increased remarkably. Methanol-eluted samples of blister fluid used for the radio-immunoassay showed the same chemotactic activity. In addition, the dermis developed marked edematous changes. On the basis of the results presented above, LTB4 and i-LTC4 appear to be generated in skin lesions of BP, the former attracting PMNL to the dermis and the latter causing exudation of blister fluid. Both compounds directly enhance the exacerbation of BP skin lesions. PMID- 2555605 TI - [A case of mucinous carcinoma of the skin]. AB - We were consulted by an 81-year-old man who had been complaining of a slow growing tumor on his abdomen for 20 years. Histologically, small islets of the tumor cells were floating in mucinous lake separated by fibrous septa, so we diagnosed this tumor as mucinous carcinoma of the skin (Mendoza). Electron microscopically, nuclei of the tumor cells were slightly folded. There were a few secretory granules and many secretory vacuoles in cytoplasm, but no findings of decapitation secretion. Like some other authors' reports, our data suggest that this tumor has an eccrine gland origin. To our knowledge, this is the 12th case of mucinous carcinoma of the skin in the Japanese literature. PMID- 2555606 TI - [Atypical myrmecia--on the importance of the anatomical site of HPVs-infection as a morphological factor]. AB - An 11-year-old Japanese girl developed a horny tumor branching into three parts in the fourth interdigital fold of the left hand; histologically it had numerous eosinophilic granular inclusion bodies. A histological diagnosis of myrmecia was made, which was further confirmed by the demonstration of human papillomavirus type 1 (HPV-1) by the in situ hybridization method. However, the clinical picture did not resemble the so-called myrmecia. A review of other atypical cases of myrmecia on areas other than the palms and soles may indicate the role of topical factors in determining the shape of viral warts. The concept of myrmecia should be revised in view of the additional information that there is histological heterogeneity in the cytoplasmic inclusion bodies, which are the basis of the definition of myrmecia. PMID- 2555607 TI - [Possible mechanisms of diethyldithiocarbamate-induced duodenal mucosal damage in rats]. AB - Diethyldithiocarbamate (DDC), which markedly lowers activity of Cu, Zn-superoxide dismutase (SOD), induces not only gastric but also duodenal mucosal damage in rats. In this experiment, we studied on the mechanisms of DDC-induced duodenal mucosal damage with special reference to aggressive and defensive factors. Possible importance of oxygen-derived free radicals was also investigated. Male Wistar rats weighing about 200 g received a single subcutaneous injection of DDC at a dose of 1 g/kg. Mucosal lesions such as edema, shortness of villi and shallow erosion developed from 3 hours after DDC injection. The ulcer index reached at its maximum 12 hours later. Gastric acid output was markedly inhibited from 87.7 +/- 8.7 to 14.0 +/- 2.6 microEq/kg/h (m +/- SE) 30 minutes after the administration of DDC (p less than 0.01), which gradually recovered thereafter. Duodenal mucosal blood flow before DDC injection was 144.7 +/- 12.7 ml/min/100 g, which decreased to 83.3 +/- 5.1 (p less than 0.01) 1 hour later. Transmucosal potential difference of the duodenum before DDC injection was -9.5 +/- 1.0 mV, which decreased to -3.8 +/- 1.3 (p less than 0.05) 30 minutes later. Duodenal alkaline secretion before DDC injection was 15.6 +/- 0.8 microEq/kg/h, which decreased to 5.0 +/- 0.9 microEq/kg/h (p less than 0.01) 90 minutes later. PAS stained mucus of Brunner's glands remained unchanged by the administration of DDC. SOD activity was markedly inhibited from 57.7 +/- 4.0 to 15.7 +/- 0.9 u/mg protein 3 hours after the administration of DDC (p less than 0.01), then recovered 24 hours later.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555608 TI - [Remission state and five-year survival after transcatheter arterial embolization therapy in patients with hepatocellular carcinoma]. AB - Repeated transcatheter arterial embolization (TAE) therapy was done for 138 cases with hepatocellular carcinoma. Relationship was investigated between long term survival more than 3 years and tumor necrosis rate estimated on radiological images (superselective angiogram and/or computerized tomography) after therapy. Excellent tumor necrosis (radiological disappearance of viable tumor) was attained in 35 out of 138 cases through repeated TAE. When "remission" is defined as excellent tumor necrosis and normal alphafetoprotein value lasting for more than one year, remission state was attained in 17 cases (12.3%). Cumulative three year survival rate was 37.0%, and 5-year survival was 26.5%. Out of 37 three-year survivors, only 12 cases (32.4%) experienced remission state in their clinical course. On the other hand, 7 cases (77.7%) of 9 five-year-survivors experienced remission at least one time. Though good tumor necrosis and 3-year survival were achieved by only performing repeated TAE therapy, "remission" state was important and necessary to get 5-year survival. PMID- 2555610 TI - [A case of small cell carcinoma of the pancreas]. PMID- 2555609 TI - [A case of resection of small liver cancer, diagnosed only by ultrasonically guided liver biopsy]. PMID- 2555611 TI - [Analysis of subjective symptoms of coldness and numbness in the upper and lower limbs among patients with vibration syndrome]. AB - Coldness and numbness of the lower limbs in patients with vibration syndrome were studied in correlation with the frequency of attacks of vibration-induced white finger (VWF), coldness and numbness of the upper limbs, and awakening at night due to numbness of the arm. Subjects were 229 patients aged 50-69 years without disease other than vibration syndrome who had mainly operated chain saws. The present study showed that patients with more frequent VWF attacks had a higher prevalence and frequency of coldness in the fingers, awakening at night due to numbness of the arm, and coldness and numbness in the legs. Particularly patients having almost daily attacks of VWF had a high prevalence of these symptoms. Positive correlation was observed in prevalence and frequency between coldness of the fingers and that of the legs, and between coldness and numbness in the legs. The prevalence of VWF, numbness and coldness of the fingers, and coldness of the legs was higher the longer the total chain saw operating period. The present findings suggest that patients with severe circulatory disturbances in the upper limbs have severe circulatory disturbance in the lower limbs. Further studies on circulatory disturbances in the lower limbs are indicated, and their mechanism involved should be also pursued from the pathogenetic point of view. PMID- 2555612 TI - [Pyrimidine nucleotides accumulated in erythrocytes of lead workers]. PMID- 2555613 TI - [Prostaglandin E2 receptor and GTP binding protein in the normal and nephrotic rat kidney]. AB - The relationship between prostaglandin E2 (PGE2) receptor and GTP binding protein was studied in the normal and aminonucleoside-induced nephrotic rats. Affinities and maximal binding sites of PGE2 receptors in renal medulla of the male Wistar rats were examined by Scatchard analysis. Cyclic AMP and inositol phosphates were measured in the medulla tissues to examine the intracellular mediators of PGE2 receptor activation. The following results were obtained: 1) GTP gamma S, a nonhydrolyzable guanine nucleotide, increased the 3H-PGE2 binding in a concentration-dependent manner, and pertussis toxin partially attenuated the GTP gamma S effect, 2) PGE2 stimulated the accumulation of inositol phosphates in the medullary tissues with a similar dissociation constant (Kd) value of binding analysis, and high concentrations of PGE2 stimulated the accumulation of cyclic AMP, 3) the affinity of PGE2 receptors in the renal cellular membranes taken from nephrotic rats were significantly lower than that in normal rats, and the effects of GTP gamma S on the affinity was greater in nephrotic rat kidney than normal one. These results suggest that PGE2 receptor of the rat kidney links to a GTP binding protein. In nephrotic rat kidney, damages occur not only in PGE2 receptor but also in the GTP binding protein. PMID- 2555614 TI - [The role of the Na, K-ATPase inhibitor in renal sodium handling in patients with essential hypertension]. AB - The present study aimed to elucidate the role of Na, K-ATPase inhibitor in renal sodium metabolism in essential hypertension. Mean arterial pressure (MAP), heart rate(HR), urine volume (UV), urinary excretion of sodium (UNaV), endogenous creatinine clearance (Ccr), fractional excretion of sodium (FENa), plasma renin activity(PRA) plasma aldosterone concentration(PAC), plasma noradrenaline concentration (PNA) and urinary excretion of noradrenaline(UNA) were measured before and after intravenous injection of ouabain (0.1 mg/m2.BSA) in 12 normotensive(NT) and 22 mild-to-moderate essential hypertensive subjects(EHT). Following ouabain injection, UV, UNaV FENa significantly increased, but PRA decreased, in both NT and EHT. MAP, HR, Ccr, PNA, and UNA did not change significantly in either group. On the other hand, a significant decrease in PAC was observed in NT, but not in EHT. The changes of UNaV and FENa were significantly attenuated in EHT as compared to NT. No significant difference in change of MAP, HR, UV, Ccr, PNA, UNA, or PRA was demonstrated between NT and EHT. A significantly positive correlation was found between delta UNaV and delta FENa in both NT and EHT, while no significant correlation was observed between delta UNaV and delta MAP, delta UV, delta Ccr, delta PRA, delta PAC, delta PNA and delta UNA in either group. These results suggest that 1) Na, K-ATPase inhibitor clearly augments natriuresis by suppression of sodium reabsorption in renal tubules, 2) since this augmentation was attenuated, there is an elevation of endogenous Na, K-ATPase inhibitor(s) should be considered in EHT, and 3) an increase of the inhibitor might participate to the hypertensive mechanism in EHT. PMID- 2555615 TI - [A case of pulmonary tuberculosis in a long-term survivor with small cell carcinoma of the lung]. AB - A 64-year-old woman complained of chest pain, back pain and dry cough. Chest X ray film showed marked left hilar enlargement and left pleural effusion. Biopsy of the bronchial mucosa and demonstrated small cell carcinoma of the lung. Treatment with cisplatin, adriamycin and etoposide led to regression of symptoms and chest X-ray findings within 3 months. She received maintenance chemotherapies at the outpatient clinic for 2 years after the first therapy. Thirty-months after the first admission, chest X-ray film showed multiple small nodules in the left upper lobe. Chest tomography and high-resolution computed tomography showed acinar or lobular nodules disseminated in the left upper lobe. Mycobacterium tuberculosis was obtained from bronchoalveolar lavage fluid of the left upper lobe. Pulmonary tuberculosis in a long-term survivor of small cell carcinoma of the lung is very rare. PMID- 2555616 TI - Spread of the single spike activity in the myenteric plexus of the guinea-pig duodenum. AB - The spontaneous single spikes in the myenteric plexus of the guinea-pig duodenum were recorded extracellularly. The myenteric plexus neurones in the cardiac region of the stomach also exhibited the spontaneous single spike activity. The frequency of these spikes was at low level. By the treatment with a high Mg and Ca-free Krebs solution, the single spikes were abolished. The generation of the single spikes was also inhibited by d-tubocurarine. These results suggest the presence of nicotinic neurotransmission in the myenteric plexus. The field stimulation could not evoke the single spike activity. The single spike activities of the myenteric plexus neurones were recorded by two electrodes situated in a ganglion or at two different ganglia. The simultaneous recordings by two different spots showed that there were corresponded and non-corresponded spikes. The former may be due to the conduction of excitation. The values of the correspondence ratio were calculated and the values ranged from 39.1% to 69.6% in a ganglion and ranged from 55.2% to 94.2% in different ganglia. The single spikes may spread over 10 mm in a longitudinal direction. These results suggest that the intra- and interganglionic interactions appear within the myenteric plexus and ganglia are interconnected into a functional integrative circuitry. PMID- 2555617 TI - [Bladder carcinoma in situ which harbored human papilloma virus. A case report]. AB - Last year, we reported that human papilloma virus type 16 genome (HPV 16 genome) was detected in a case (S.Y.) of bladder carcinoma in situ (bladder CIS) (Cancer Res., 1988). Since then, a number of bladder tumors other than CIS were searched for HPV genome. However, no HPV genome was detected in the bladder tumors. From the results, we consider that HPV may not have a relation with all types of bladder tumor but with only a part of it. In the current report, the case (S.Y.) is presented more precisely than before, in particular on the characteristic bladder lesion. The patient was a 40-year-old female with immunodeficiency and anemia who was referred from a hospital with a complaint of asymptomatic pyuria. Cystoscopic examination revealed a bladder tumor, well-demarcated, white and velvety lesion with slight elevation. On November 25, 1987, she underwent total cystectomy, resection of the anterior vaginal wall and of a part of vulval skin, and ileal conduit formation. Postoperative course was stormy because of bleeding from the wounds and thrombophlebitis in the right femoral vein. In spite of the episodes, she eventually recovered and was discharged 2 months later. However she was readmitted 9 months later due to severe anemia which was ascribed to acute myelogenous leukemia. She is now on cancer chemotherapy for leukemia. PMID- 2555618 TI - The microheterogeneity of bovine serum alpha 1-acid glycoprotein. AB - The microheterogeneity in bovine serum alpha 1-acid glycoprotein (alpha 1AGP) was examined by crossed affino-immunoelectrophoresis with concanavalin A(Con A). In healthy cows, serum alpha 1AGP revealed 2 fractions which were named fraction 1 and 2 according to the degree of binding affinity to Con A. The mean +/- SD of serum alpha 1AGP concentration was 0.31 +/- 0.09 g/l and the relative amounts of fraction 1 and 2 were approximately 67% and 33% respectively on this method. In bovine leukemia cases, 5 out of 8 had a high serum alpha 1AGP concentration of more than 0.7 g/l. On the electrophoretic pattern, the other 2 fractions (Con A weakly and non-reactive fractions, named fraction 3 and 4) were observed in 5 cases and their relative amounts of 4 fractions were 17.8-28.5%, 35.5-46.0%, 17.9 24.4% and 12.0-17.2%, respectively. In bovie leukemia virus infected cows with no clinical symptoms, however, serum alpha 1AGP concentration and the electrophoretic patterns were almost the same as those in healthy cows. PMID- 2555619 TI - Pathological findings of two types of lymphoid malignancy in sheep inoculated with bovine leukemia virus. AB - Different types of lymphoid malignancy were observed in two sheep inoculated with BLV-containing materials. Sheep 1 showed severe leukemic change in the peripheral blood and splenomegaly but lymphosarcoma in the lymph nodes was absent. Sheep 2 had lymphosarcoma in the lymph nodes and various organs. Neoplastic cells had B cell marker in both cases and a few neoplastic cells contained intracytoplasmic IgM in sheep 2. It was presumed that B-cells might be transformed into neoplastic cells on the way of their differentiation. Some of neoplastic cells might have ability of immunoglobulin-production in sheep 2. PMID- 2555620 TI - Neutralizing antibody to three different serotypes of porcine rotavirus in pigs. PMID- 2555621 TI - Experimental infection of pigs with porcine hemagglutinating encephalomyelitis virus. PMID- 2555622 TI - Enhanced pathogenicity of chicken anemia agent by infectious bursal disease virus relative to the occurrence of Marek's disease vaccination breaks. PMID- 2555623 TI - [The kinin and blood coagulation systems in patients with myocardial infarction during rehabilitation]. AB - In 161 patients with acute myocardial infarction, the kallikrein-kinin system was found to be activated. In addition, there were alterations in indices of the coagulative system, which were suggestive of hypercoagulation. The unidirectional changes in the indices of the systems in question during therapy in the periods of rehabilitation are attributable to the common character of trigger mechanisms responsible for their activation and to the involvement of the kinin system in the regulation of hemostatic potential. PMID- 2555624 TI - [Abrikosov's endobronchial myoblastic myoma]. PMID- 2555625 TI - [Locoregional chemotherapy]. PMID- 2555626 TI - [Fulminant gas gangrene panophthalmia following perforating scleral injury]. AB - A 13-year-old boy developed Clostridium perfringens panophthalmitis due to a penetrating scleral injury. The characteristic symptoms were unusually severe periocular pain, lid edema, decreased motility and protrusion of the eye, severe conjunctival chemosis, pus covering the conjunctiva and cornea, green-brown hypopyon, gas bubbles in the anterior chamber, and amaurosis within 60 hours. In view of the patient's ocular status and impaired general condition enucleation was unavoidable. The results of laboratory workups are reported in addition to the clinical and ophthalmologic findings. PMID- 2555627 TI - Unusually rapid development of a HBsAG-positive liver cirrhosis after liver transplantation. AB - We report the case of a 44-year-old man who was transplanted in 1986 for hepatocellular carcinoma in a HBsAG-positive liver cirrhosis. The patient had no severe complications postoperatively. He received passive immunization for the prevention of hepatitis B reinfection during the first 6 months after liver grafting. Twelve months after the transplantation the new liver was reinfected with hepatitis B virus. Without any clinical or laboratory signs of severe hepatitis, the patient developed a histologically proven complete liver cirrhosis within 8 months after reinfection of the graft. The reasons for this might have been, first, a deleterious course of the infection under immunosuppressive therapy, and, second, the additional influence of a postoperatively acquired CMV infection or the combined toxic influence of cyclosporin A and its metabolites on the acute inflammation in the liver. PMID- 2555628 TI - Gel chromatographic characterization of immunoreactive adrenocorticotropin in patients with ACTH hypersecretion. AB - We investigated the molecular size of circulating immunoreactive ACTH by gel chromatography in patients with ACTH hypersecretion due to various disorders of the hypothalamic-pituitary-adrenal axis. 4 patients with Addison's disease, 2 with Nelson's syndrome, 4 with Cushing's disease, 6 with the ectopic ACTH syndrome (2 bronchial carcinoma, 1 medullary carcinoma, 1 metastatic islett cell carcinoma, 1 benign bronchial carcinoid and 1 patient with occult ectopic Cushing's syndrome) and 1 patient with hypersecretion of ACTH from a clinically nonfunctioning pituitary adenoma were studied. Analysis of the molecular size of immunoreactive ACTH was performed by gel chromatography on a Sephadex G-75 column (superfine, 100 x 1.5 cm) equilibrated with 1% formic acid. 2 ml fractions were collected and evaporated to dryness. The ACTH content of the recovered samples was determined by RIA. In Addison's disease, Nelson's syndrome and Cushing's disease the plasma showed a single peak of ACTH immunoreactivity at the expected position of 1-39 ACTH. In the ectopic ACTH syndrome the plasma of 4 patients revealed at chromatography at least one other peak eluting between the void volume and 1-39 ACTH suggestive of a high molecular weight form of ACTH whereas plasma of 2 patients showed only a single ACTH peak at the position of labeled 1 39 ACTH. The patient with a clinically non-functioning pituitary adenoma revealed a gel filtration pattern similar to the patients with ectopic ACTH syndrome and secretion of high molecular weight ACTH. We conclude that secretion of high molecular weight forms of ACTH is not a unique feature of the ectopic ACTH syndrome.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555630 TI - [Blood levels of pituitary gonadotropins, sex hormones, gastrin and cAMP in duodenal ulcer]. AB - Radioimmunoassay was employed to measure blood levels of LH, FSH, testosterone, estradiol, progesterone, estriol, gastrin and cAMP in 184 duodenal ulcer patients (141 males and 43 females) and 81 controls (40 males and 41 females). For duodenal ulcer patients hypophyseal gonadotropic function was activated while the blood picture of sex hormones is sex-related. Functional balance between the study hormones typical for healthy subjects in duodenal ulcer was found impaired. Derangement of hormonal homeostasis of hypophyseal-gonadal system of the blood and its functional relations with gastroduodenal system may be a contributing factor in pathogenesis of peptic ulcer. PMID- 2555629 TI - [Diagnosis and clinical aspects of gastrointestinal cytomegalovirus diseases in patients with human immunodeficiency virus 1 infection]. AB - Eleven Human Immunodeficiency Virus 1 (HIV1)-infected patients (10 male, 1 female; age 23-51 years (median 36); 10 male homosexuals, 1 IV drug abuser; WR3 1 patient, WR5 5, WR6 5) with intestinal Cytomegalovirus (CMV)-manifestations were compared with a group of 78 HIV1-infected patients in respect to their clinical, immunological and virus-serological data and the results of the histological and microbiological examination of endoscopically obtained biopsies. No differences were observed on age, sex, risk of infection, stage and immunological status. Bloody diarrhea was most important in discriminating CMV-colitis and non-CMV related intestinal manifestations. Dysphagia and other symptoms occurring in patients with CMV-esophagitis were not able to predict CMV-esophagitis specifically. 6 of 11 patients with serological findings consistent with an active CMV-infection had no detectable CMV-manifestations; 6 of 11 patients with intestinal CMV-manifestations did not show serological findings suggestive of active CMV-infection. Ulcerative alternations of intestinal mucosa represent the most powerful indicator of intestinal CMV-disease in endoscopical examination. Only in two patients, ulcerative alterations were seen without diagnosis of CMV disease being established. CMV was isolated in one of 11 patients, in two patients CMV was isolated from biopsies of unchanged mucosa. Simultaneous infection by HSV and CMV was detected in three patients, in one patient in the same localisation. Histology revealed inclusion bodies in 8 of 11 patients with intestinal CMV-disease, in no case inclusion bodies were seen without CMV disease. PMID- 2555631 TI - [Reaction of osseous tissue and the osteoclast population in rats to biphosphonates and vitamin D3 under conditions of hypokinesia]. AB - Oral administration of hydroxy-dimethyl aminopropylene biphosphonate (DMAP) at a dose of 0.5 mg/kg increased the volume density (mass) of trabecular bone in the tibial metaphysis of untreated rats and helped to maintain it at the normal level in hypokinetic rats. Oral administration of hydroxy-ethylidene biphosphonate (OEDP) at a dose of 20 mg/kg and vitamin 24,25(OH)2D3 at a dose of 1,25 micrograms did not induce any quantitative changes in bones of untreated or immobilized rats. The combined treatment with either biphosphonate and vitamin D3 exerted a similar effect as compared to that of biphosphonates used singly. During hypokinesia the total amount of osteoclasts declined. This effect was also seen in response to biphosphonates and vitamin D3. The two biphosphonates produced an opposite effect on the osteoclast population in untreated animals: it increased in response to OEDP and decreased in response to DMAP. In contrast to OEDP, low doses of DMAP can efficiently inhibit bone resorption upon various routes of its administration and can be viewed as a preventive and therapeutic drug in the case of osteoporosis. PMID- 2555632 TI - Neutrophil function and lipid peroxidation in a rat model of multiple organ failure. AB - Multiple organ failure (MOF) was induced by sterile intraperitoneal inoculation of zymosan in the rat. This results in a typical triphasic illness with maximal clinical signs at Days 2 and 14. In this study, granulocyte superoxide production (unstimulated and phorbol myristic acid stimulated) was studied as well as lipid peroxidation (TBAR) in plasma, liver, and lung tissue. Mainly TBAR levels in liver and lung tissue closely correlated with the triphasic clinical illness, while bacteriological data did not. It is concluded that the severe inflammatory response in this experimental model probably is the result of excessive toxic oxygen radical production. The first phase of illness may mainly be due to oxygen radical formation by activated PMN, the third phase of illness to the production of lysosomal enzymes (proteinases) from PMN, and activated macrophages as indicated by elevated N-acetylglucosaminidase levels. PMID- 2555633 TI - The t(2;5)(p23;q35): a recurring chromosomal abnormality in Ki-1-positive anaplastic large cell lymphoma. AB - In lymphoid neoplasms, nonrandom cytogenetic abnormalities correlate with clinical, morphologic and immunophenotypic features. A subtype of non-Hodgkin's lymphoma, which expresses the Ki-1 antigen (CD30) and has distinct morphologic and clinical features, has recently been described. We now report the association of a reciprocal translocation involving the short arm of chromosome 2 (band p23) and the long arm of chromosome 5 (band q35), t(2;5)(p23;q35), with Ki-1 positive anaplastic large cell lymphoma. Rearrangement of the genes that are located at the breakpoints on chromosomes 2 and 5 may be a critical step in the pathogenesis of this lymphoma. PMID- 2555634 TI - Expression of c-fgr in EBV positive and negative B cell tumors. PMID- 2555635 TI - Further evidence for the GABAergic influence on memory. Interaction of GABAergic transmission with angiotensin II on memory processes. AB - In experiments on male Wistar rats trained on active avoidance (shuttle-box) and passive avoidance (step-through) tasks, we found that (-) nipecotic acid, the inhibitor of GABA reuptake, and muscimol, a GABAA-receptor agonist, applied after training either improved or had no effect on retention, depending on the dose used. Angiotensin II (ATII) at a dose of 0.1 microgram/kg injected intracerebroventricularly (i.c.v.) after training facilitated retention. Combinations of ATII and (-) nipecotic acid or muscimol potentiated the memory effects of ATII and GABAergic drugs, respectively. Blockade of GABA receptors (GABAA) by bicuculline or picrotoxin abolished the effects of GABAergic agonists on ATII. It is suggested that GABAergic transmission participates in the consolidation and formation of memory traces and in the retention-facilitating mechanism of action of ATII. PMID- 2555636 TI - Proteolytic activation of aldosterone biosynthesis in rabbit adrenal capsular cells by alpha-chymotrypsin. AB - The effect of chymotrypsin on aldosterone biosynthesis by dispersed rabbit adrenal capsular cells was examined. Bovine alpha-chymotrypsin at concentrations of 10(-7) to 10(-5) M stimulated aldosterone production, and human chymotrypsin had an even stronger stimulatory effect. Bovine trypsin had no effect on aldosterone production by adrenal cells. Chymotrypsin treatment did not change the sensitivity of the adrenal cells to ACTH or angiotensin II. These results suggest the existence of unique chymotrypsin-susceptible sites on rabbit adrenal capsular cells, the digestion of which results in stimulation of aldosterone biosynthesis. PMID- 2555637 TI - Nucleoside analogs as non-substrate inhibitors of herpes simplex viruses thymidine kinase. AB - We have examined the capacity of a series of 6-substituted pyrimidine and 2 substituted purine derivatives to inhibit mammalian thymidine kinase and the thymidine kinases encoded by type 1 and type 2 herpes simplex viruses. Several N2 substituted guanine and deoxy guanosine derivatives displayed selective inhibitory activity against the HSV-1 and HSV-2 thymidine kinases by competing with the phosphorylation of thymidine, suggesting a possible novel pharmacological approach to herpes viruses infections. PMID- 2555638 TI - N-isobutyryl-His-Trp-Ala-Val-D-Ala-His-Leu-NHMe (ICI 216140) a potent in vivo antaconist analogue of bombesin/gastrin releasing peptide (BN/GRP) derived from the C-terminal sequence lacking the final methionine residue. AB - The GRP receptor mediated growth response in Swiss 3T3 cells has been used to identify BN/GRP antagonists. Analysis of bombesin antagonism by substance P analogues and by truncated GRP analogues revealed that deletion of the C-terminal methionine residue was important for antagonism. Des-Met analogues showing potent antagonist activity in the in vitro 3T3 system (IC50 approximately 2nM) were synthesized. Further structural modification of these peptides led to the identification of (CH3)2CHCO-His-Trp-Ala-Val-D-Ala-His-Leu-NHCH3 (ICI 216140) which reduced bombesin-stimulated rat pancreatic amylase secretion to basal levels when administered subcutaneously at 2.0 mg per kg. PMID- 2555639 TI - Hypertonic sodium bicarbonate partially reverses QRS prolongation due to flecainide in rats. AB - Hypertonic (1M) sodium bicarbonate can partially reverse the cardiac toxicity of some Class IA antiarrhythmic agents, presumably by antagonizing sodium channel inhibition. We studied the effects of 1M sodium bicarbonate on toxicity due to the Class IC drug flecainide. Anesthetized rats received i.v. loading and maintenance doses of flecainide to produce QRS prolongation of 76% that was stable over the 60 min study period. 20 min after the start of the maintenance infusion, groups of 8 rats received an i.v. infusion of 1M sodium bicarbonate (6 meq/kg) or an equal volume of 0.9% saline. QRS prolongation was reduced by 1M sodium bicarbonate but not only 0.9% saline (% change -12.2 +/- 10.0 v. +3.0 +/- 2.7, p = 0.001). Expressed as a percent of the flecainide-induced QRS prolongation, bicarbonate reduced this prolongation by 26.5%. The improvement in QRS duration persisted until sacrifice 30 min later. Compared to controls, the bicarbonate group had a significantly higher blood pH (7.55 +/- 0.06 v. 7.44 +/- 0.05) and serum sodium concentration (149 +/- 5 v. 137 +/- 2 meq/l). Serum flecainide concentrations were similar. These data suggest that 1M sodium bicarbonate can partially reverse flecainide-induced conduction delay in rats. This effect may be due to changes in the extracellular pH and sodium concentration. 1M sodium bicarbonate may be useful in assessing the role of sodium channel inhibition in mediating the toxicity of flecainide or other Class IC drugs. PMID- 2555640 TI - Differential effects of naloxone on basal and stress-induced release of ACTH and prolactin in the male rat. AB - The effect of i.v. injection of various doses of naloxone (NAL) on plasma adrenocorticotropin (ACTH) and prolactin (Prl) in conscious animals bearing an indwelling intrajugular catheter was assessed. The effects were evaluated in animals which were left undisturbed and in others subjected to either restraint or ether stress. The results revealed that the dose of 3 mg/kg of NAL significantly reduced basal Prl levels, whereas a dose of 6 mg/kg of NAL was required to block completely either ether or restraint stress-induced release of Prl. The behavior of ACTH contrasted with that of Prl. There was no effect whatsoever of the 3 mg/kg dose of NAL on either resting or stress-induced ACTH levels, whereas a 6 mg/kg or 12 mg/kg dose of NAL elevated resting ACTH levels and only partially attenuated the further elevation induced by stress in these animals. The results clearly indicate a NAL sensitive step in the control of resting and stress-induced Prl release but indicate that the control of resting and stress-induced release of ACTH is different in that the predominantly millimicron receptor blocker, NAL, can elevate ACTH at high doses and can only partially block the response to stress. In contrast to Prl where opioid peptide control is solely stimulatory, this control of ACTH secretion appears to have both stimulatory and inhibitory features. PMID- 2555641 TI - Differential development of acute tolerance to analgesia, respiratory depression, gastrointestinal transit and hormone release in a morphine infusion model. AB - To determine whether the differences in development of acute tolerance to several morphine actions correlate with the mu receptor subtype mediating them, we have examined the appearance of acute tolerance to analgesia, respiratory depression, gastrointestinal transit, and hormone release in an intravenous morphine infusion model. Analgesia, a naloxonazine-sensitive mu1 action, peaked at 2 hr after initiation of the infusions. The log dose-response relationship of the infusion rate to peak tailflick latency was linear from 10 to 50 micrograms/kg/min. By 8 hr, the tailflick latencies declined nearly to baseline levels, implying the rapid development of tolerance. Tolerance to morphine-induced prolactin release, another mu1 action, also developed rapidly over 8 hr. In contrast two mu2 actions, respiratory depression measured with arterial blood gas, determinations and gastrointestinal transit, showed no significant tolerance over a similar 8 hr infusion. We also observed no tolerance to morphine-induced growth hormone release, a non-mu1 action, over the same period. Thus, these results demonstrate that mu1 actions develop tolerance in an infusion model far more rapidly than a number of naloxonazine-insensitive (non-mu1) ones and may help explain differences in the rate of tolerance development to morphine actions. PMID- 2555642 TI - Tissue plasminogen activator and plasmin independently decrease human neutrophil activation. AB - Tissue-plasminogen activator (t-PA) and plasmin both decrease platelet aggregation, which may contribute to thrombolysis and tissue salvage. Since neutrophils may contribute to reperfusion injury, we examined the effects of t-PA and plasmin on human neutrophil function. t-PA (1 to 100 micrograms/ml) decreased f-MLP-induced chemotaxis and ionophore A23187-induced superoxide and LTB4 release in isolated neutrophils, and these effects were not blocked by the plasmin inhibitor epsilon-aminocaproic acid (epsilon-ACA). On the other hand, plasmin (0.05 to 0.5 units/ml) also decreased these neutrophil functions but its effects were blocked in the presence of epsilon-ACA. Thus, while both t-PA and plasmin decrease neutrophil functions, effects of t-PA are independent of plasmin generation. Cumulative effects of t-PA and plasmin on neutrophil functions may relate to the overall efficacy of t-PA in thrombotic disorders. PMID- 2555643 TI - The effects of althesin on luteinizing hormone release cannot be explained by actions of the GABAA receptor alone. AB - Althesin and pentobarbitone are anaesthetics which act by prolonging the open time of the chloride channels of the GABA(A) receptor. To explain why luteinizing hormone (LH) release is less depressed by Althesin anaesthesia than by pentobarbitone anaesthesia we suggest that either Althesin is a less potent anaesthetic in the preoptic area or that Althesin as well as stimulating GABA(A) receptors has some other action, perhaps stimulation of GABA(B) receptors, which may facilitate LH release. To investigate the relative potency of the anaesthetics in the preoptic area nine cats were anaesthetised, six with Althesin and three with pentobarbitone, mounted in a stereotaxic frame and prepared for extracellular recording and stimulation of spontaneously active units in the preoptic region. When cats anaesthetised with Althesin were compared with cats anaesthetised with pentobarbitone there were significantly fewer of these units for the number of tracks made. These units also had a significantly lower frequency and a distribution significantly skewed toward lower frequencies. Electrical stimulation of the fornix and of sites in the medial basal hypothalamus and medial forebrain bundle inhibited about 50% of the units and the median duration of the inhibitory pause was significantly longer following stimulation at all three sites in cats anaesthetised with Althesin. We conclude that Althesin is a more potent anaesthetic than pentobarbitone in the preoptic region and that its effects on LH release cannot be explained by its effects on the GABA(A) receptor alone. PMID- 2555644 TI - Lipid characteristics of RSV-transformed Balb/c 3T3 cell lines with different spontaneous metastatic potentials. AB - To determine whether a metastatic phenotype may be correlated with a characteristic lipid pattern, we compared the lipid composition of low metastasizing Balb/c 3T3 cells transformed by the B77 strain of Rous sarcoma virus (B77-3T3 cells) with that of a subclone isolated by growth in 0.6% agar, the B77-AA6 cells, which exhibit a high capacity for spontaneous metastasis. B77 3T3 cells revealed characteristics in their lipid composition common to other systems of transformed cells, i.e., an accumulation of ether-linked lipids, a reduction of the more complex gangliosides, an increase of oleic acid (18:1) and a decrease of arachidonic (20:4) and C22 polyunsaturated fatty acids in phospholipids. High metastatic B77-AA6 cells showed: a) an even more marked decrease of complex gangliosides; b) a more pronounced increase of 18:1 and decrease of 20:4 and 22 polyunsaturated fatty acids in certain phospholipid classes; and c) a higher percentage of alkyl-acyl subfractions in both phosphatidylcholine and phosphatidylethanolamine than B77-3T3 cells. Comparing the data for other systems of metastatic cells with those of lipid studies of spontaneously metastasizing B77-AA6 cell system leads us to conclude that the metastatic phenotype is characterized by a change in ether-linked lipids, rather than in fatty acids. PMID- 2555645 TI - Effect of hyperpnea on enzymes of the CDPcholine path for phosphatidylcholine synthesis in rat lung. AB - We have examined the activity of three enzymes in pulmonary surfactant phosphatidylcholine synthesis following the hyperpnea induced by having rats either inspire 5%CO2/13%O2/82%N2 for 24 hr or swim in thermoneutral water for 30 min. Both stimuli markedly increase frequency and tidal volume of breathing and promote the release of surfactant. Lungs were perfused to remove blood, lavaged, and then homogenized in 1 mM Hepes, 0.15M KCl at pH 7.0. The homogenate was centrifuged at 9,000 g (av) for 10 min to sediment the mitochondria and lamellar bodies and at 100,000 g (av) for 60 min to obtain the microsomal and cytosol fractions. Incubations were carried out under determined optimal conditions and zero order kinetics. Choline kinase (CK), cholinephosphate cytidylyltransferase (CP-cyT) and choline phosphotransferase (CPT) were assayed by the incorporation of [methyl-14C]choline chloride into phosphocholine, [methyl-14C]phosphocholine into CDPcholine, and [14C]CDPcholine into phosphatidylcholine, respectively. The incubation products were separated by thin-layer chromatography. Whereas both forms of hyperpnea increased the activity of CP-cyT in the microsomal fraction, they had no effect on the activity of either cytosolic CP-cyT and CK, or microsomal CPT. A similar increase in tidal volume in an isolated perfused rat lung had no effect. We conclude that, in vivo, hyperpnea increases the activity of CP-cyT, the rate-limiting enzyme in phosphatidylcholine synthesis. Whether this is due to an increase in the amount of enzyme, or of a cofactor, is unknown. PMID- 2555646 TI - Effect of magnesium deficiency on delta 6 desaturase activity and fatty acid composition of rat liver microsomes. AB - Experimental Mg2+ deficiency was induced in a group of rats by feeding them a Mg2+-deficient diet for 23 days. They were pair-fed to compare with a control group of rats fed a Mg2+-sufficient diet. In the Mg2+-deficient group the plasma total cholesterol and triglyceride levels were increased while HDL-cholesterol was decreased. In the Mg2+-deficient group the plasma level of thiobarbituric acid reacting substances (TBARS) used as a measure for lipid peroxidation was increased. The increase was attributed to the increased cytosolic Ca2+ in Mg2+ deficiency which can cause: 1) increase of hydro and endoperoxide levels as a consequence of the increase of arachidonic acid release and eicosanoid synthesis in Mg2+-deficiency, and 2) inhibition of the mitochondrial respiratory activity and activation of Ca2+-dependent proteases which may activate the conversion of xanthine dehydrogenase to xanthine oxidase which generates active O2 species. In the Mg2+-deficient group, the fatty acid composition of the liver microsomes indicated a slower rate of conversion of linoleic acid to arachidonic acid which was consistent with the decrease of delta 6 desaturase activity in liver microsomes of Mg2+-deficient rats as measured in vitro. The decrease of delta 6 desaturase activity was attributed to the lower concentration of actual enzyme molecules as a result of the decreased rate of protein synthesis in Mg2+ deficiency. The possible effects of the increased catecholamine release in Mg2+ deficiency are discussed. PMID- 2555648 TI - Effect of eicosapentaenoic acid ethyl ester on proteinuria of streptozotocin induced diabetes mellitus in rats. AB - Streptozotocin (45 mg/kg) was intravenously administered to 7-week-old Wistar rats through their tail veins. After 11 days, the rats were divided into two groups. One group was fed a lipid-free diet (90%, w/w) plus lard (8%) and safflower oil (2%) for four weeks (Diet 1 group, n = 12). The other group was fed in the same way, except that safflower oil was replaced by 90% pure eicosapentaenoic acid (EPA) ethyl ester (Diet 2 group, n = 13). Twenty-four-hour urine was collected just before the diets started and during the experiment at 7 day intervals. In the second and third week, the levels of proteinuria were significantly lower in the Diet 2 group than they were in the Diet 1 group. There was no significant difference in the levels of creatinine, urea nitrogen, or lipids in plasma or in body weights between the two groups after four weeks on the diets. Because Diet 2 reduced proteinuria of diabetic rats compared to Diet 1, an EPA-rich diet may retard the development of diabetic nephropathy. PMID- 2555647 TI - Activation of rat liver cholesterol ester hydrolase by cAMP-dependent protein kinase and protein kinase C. AB - Short term regulation of hepatic cholesterol ester hydrolase by reversible phosphorylation is described. Two different kinase systems seem to be involved in this regulation. The addition of ATP, cyclic AMP and Mg2+ to rat liver 104,000 X g supernatant (S104) produced a 100-140% increase in cholesterol ester hydrolase activity. This stimulation was abolished when protein kinase inhibitor was added prior to the addition of ATP, cyclic AMP and Mg2+. Cholesterol ester hydrolase activity was also stimulated when calcium ions, phosphatidylserine, and diolein were added to S104 along with ATP and Mg2+. Diolein in this reaction could be substituted by phorbol 12-myristate 13-acetate. Preincubation of S104 with alkaline phosphatase resulted in a deactivation of cholesterol ester hydrolase. The addition of increasing concentrations of Mg2+ to S104 produced increasing inhibition of cholesterol ester hydrolase activity, and this effect was blocked by NaF. It is suggested that rat liver cholesterol ester hydrolase is activated by cyclic AMP dependent protein kinase and protein kinase C. Deactivation is accomplished by dephosphorylation catalyzed by a phosphoprotein phosphatase, dependent on Mg2+. PMID- 2555649 TI - Occurrence of geometrical isomers of eicosapentaenoic and docosahexaenoic acids in liver lipids of rats fed heated linseed oil. AB - Mono trans geometrical isomers of 20:5 n-3 and 22:6 n-3 were detected in liver lipids of rats fed heated linseed oil. The isomers were identified as being 20:5 delta 5c,8c,11c,14c,17t and 22:6 delta 4c,7c,10c,13c,16c,19t. These fatty acids were isolated as methyl esters by preparative high-performance liquid chromatography (HPLC) on reversed phase columns followed by silver nitrate thin layer chromatography (AgNO3-TLC). The structures were identified using partial hydrazine reduction, AgNO3-TLC of the resulting monoenes, oxidative ozonolysis of each monoene band, and gas-liquid chromatography (GLC) of the resulting dimethyl esters and monomethyl esters. Fourier-transform-infrared spectrometry confirmed the trans geometry in isolated 20:5 and 22:6 isomers. The isomers of eicosapentaenoic and docosahexaenoic acids in liver lipids probably resulted from desaturation and elongation of 18:3 delta 9c,12c,15t, a geometrical isomer of linolenic acid present in the heated dietary oil. PMID- 2555650 TI - Polyphosphoinositide formation in isolated cardiac plasma membranes. AB - Phosphatidylinositol (PtdIns) and phosphatidylinositol 4-phosphate (PtdIns4P) kinase activities in plasma membranes isolated from canine left ventricle were partially characterized, and their sensitivity to a number of intracellular variables was established. PtdIns and PtdIns4P kinase activities were estimated by the formation of [32P]PtdIns4P and [32P]phosphatidylinositol 4,5-bisphosphate ([32P]PtdIns(4,5)P2), respectively, when membranes were incubated with [gamma 32P]ATP and 0.1% Triton X-100. Unlike [32P]PtdIns4P formation [32P]PtdIns(4,5)P2 formation required exogenous (PtdIns4P) substrate. [32P]PtdIns4P and [32P]PtdIns(4,5)P2 formation were insensitive to Ca2+ at concentrations ranging from 0.1-30 microM. The hydrolysis of [32P]PtdIns4P was less than 15% under standard assay conditions for measuring its formation, and was unaffected by any of the variables tested. The apparent Km of the PtdIns kinase for ATP was 53 +/- 13 (S.E.M.) microM (N = 3). ADP inhibited [32P]PtdIns4P formation competitively with respect to ATP, the Ki being 0.4 mM. The data indicate that ADP is a poor competitive inhibitor of PtdIns kinase at the concentrations which are believed to be present intracellularly normally or which may be attained during mild hypoxia provided ATP levels are maintained in the millimolar range. Hence, any response of the myocardium to alpha-adrenergic hormones during mild hypoxia would be largely unimpaired by effects of Ca2+ on PtdIns and PtdIns(4,5)P2, or of ADP on PtdIns kinase activity. PMID- 2555651 TI - Ingestion of high doses of fish oil increases the susceptibility of cellular membranes to the induction of oxidative stress. AB - Feeding rats with 4 g/kg body weight of sardine oil during 7 or 14 days increases the content of eicosapentaenoic acid and docosahexaenoic acid in the erythrocyte and hepatic microsomal membranes by 2 to 6%. These membranes show increased susceptibility to the induction of oxidative stress, expressed as lipid peroxidation, when they are exposed to Fe2+-ascorbate and to NADPH-FE3+-ADP, respectively. The results indicate that in order to prevent the increased susceptibility to lipid peroxidation, supplementation with larger amounts of antioxidants may be needed than those required to stabilize the oil. PMID- 2555652 TI - Diagnosis of hepatic hemangioma with 99mTc-labeled red cells: single photon emission computed tomography (SPECT) versus planar imaging. AB - This paper reports the results of a prospective study carried out to demonstrate the most important advantages of 99mTc-blood pool single emission computed tomography (SPECT) versus the established method of planar scintigraphy (PS) in the non-invasive diagnosis of liver hemangiomas. The study group comprised 40 patients in whom positive evidence of 56 hemangiomas, detected incidentally in the course of screening examinations, was established via SPECT and PS. The sensitivity of SPECT was 51/56 (91%), of PS 22/56 (39.3%); for lesions smaller than 30 mm in diameter, the sensitivity of SPECT was 31/38 (81.6%), of PS 6/38 (15.8%). All lesions with diameters of more than 30 mm were detected by both PS and SPECT. However, the SPECT method proved to be clearly superior in the identification of lesions which were smaller than 30 mm and located in unfavourable topographical sites (dorsal or subphrenic). The study proves that SPECT with 99mTc-labeled erythrocytes is the ideal complement to ultrasonography in the detection of liver hemangionas; its major advantage over TCT (transmission computed tomography) is its safe application in cases with contrast agent intolerance and hyperthyroid patients. Moreover, liver biopsies are dangerous and ill-suited for the verification of diagnosis. PMID- 2555653 TI - [The esophageal transport function in patients with nodular goiter and in patients operated on for thyroid cancer]. AB - A study was made of changes of esophageal transport function in 10 patients with thyroid adenoma and in 27 patients after surgical and radiotherapeutic management for thyroid cancer. The results were compared with those of a control group. Significant data on changes on esophageal transport function in patients with thyroid adenoma were unnoticed. Esophageal dysfunction after operation and radioactive iodine therapy for cancer was detected in 55%. Dysfunction was thought to be caused by dysphagia which showed direct correlation with the severity of hypothyroidism. PMID- 2555654 TI - Influence of several experimental parameters on inter and intra specific mating process efficiency. AB - In this study we investigated the influence that several experimental parameters may product on the mating process efficiency. We used the ordinary mating technique described by other authors and we examined the effect of temperature, media and plasmid size on the number of recipient cells transformed by mating procedure. The results showed that the most important parameter influencing the efficiency of the mating transformation is the media, associated with the size of the plasmid to be transferred. PMID- 2555655 TI - General characteristics and viral susceptibility of a newborn pig kidney (NPK) continuous culture. AB - We have developed a fibroblastic-like continuous culture of newborn pig kidney (NPK). The current cell line was serially passaged 160 times and appeared to be well suited for production and assay of a number of viruses affecting pigs, such as pig parvovirus, pseudorabies and transmissible gastroenteritis. The cell line appeared aneuploid, with a modal chromosome number of 36 and induced tumors, classified as fibrosarcoma, in athymic mice. PMID- 2555656 TI - Protective immunity induced by porin in experimental mouse salmonellosis. AB - The induction of protective immunity to mouse salmonellosis by porin from Salmonella typhimurium LT2 was studied. The immunization with porin induced a high level of protective immunity to salmonellosis in BALB/c mice. Mice immunized with porin exhibited significant levels of delayed-type hypersensitivity response and interleukin-2 production, indicating that porin was capable of inducing cell mediated immunity (CMI). Furthermore, we found that both T cells and sera taken from the porin-immunized mice could transfer the protection against salmonellosis into nonimmunized mice. These observations suggested that a high level of the protection to salmonellosis obtained by the porin immunization resulted from the induction of CMI in addition to humoral immunity. PMID- 2555657 TI - Effects of infection by HIV-1, cytomegalovirus, and human measles virus on cultured human thymic epithelial cells. AB - A tissue culture system for the growth of human fetal and infantile thymic epithelial (TE) cells has been established and characterized. We have investigated the effects of infection of these cells by human cytomegalovirus (CMV), measles virus, and human immunodeficiency virus type-1 (HIV-1). In the case of CMV, morphological changes were apparent by 2-4 days after viral inoculation of infantile TE cells. CMV-related antigens were detected by immunofluorescence after 12 days, and progeny infectious CMV was recovered from culture media after 18 days. Following infection by measles virus, distinctive, multinucleated giant TE cells appeared in both cultures of fetal and infantile TE cells. Measles virus-inoculated TE cells displayed an altered phenotype, as revealed by reaction with monoclonal antibodies with specificity for a variety of TE markers. Finally, infection of TE cells by HIV-1 resulted in cellular disarrangement, increased numbers of Hassall's corpuscles, and multinucleated giant cells. An increase in the number of cells reactive with monoclonal antibodies, specific for Hassall's corpuscles, was observed in the case of cells infected by either measles virus or HIV-1. These findings suggest that a variety of different viruses can successfully infect thymic epithelial tissue. Because of the important role of the thymus in development of the immune system, it is reasonable to conclude that viral infection of thymic tissue might play an important role in virus-mediated suppression of immune responsiveness. PMID- 2555658 TI - Seroepidemiology of herpes simplex virus in Yogyakarta, Indonesia. AB - Sera from 487 individuals in Yogyakarta, Indonesia, were tested for antibodies to herpes simplex virus (HSV) by a passive hemagglutination method. Age-specific incidence rates for antibodies to HSV were calculated. For sera from persons other than prostitutes, in the age group from 10 to 19, the positive rate was 48% but in the age group higher than 20, it was more than 87%. Fifty of 59 pregnant women (85%) were positive. The positive rate and the distribution of antibody levels in prostitutes were higher than in the general population. PMID- 2555659 TI - Intrahepatic arterial injections of cisplatin-phosphatidylcholine-Lipiodol suspension in two unresectable hepatoblastoma cases. AB - Two cases with unresectable hepatoblastoma were treated by intrahepatic arterial injections of cisplatin-phosphatidylcholine-Lipiodol suspension (CPLS), combined with systemic chemotherapy. Unfortunately, the first patient who was given three injections of CPLS in association with systemic chemotherapy died of progressive disease 18 months after the commencement of the therapy. However, the second patient who received about 1 year of systemic chemotherapy followed by three injections of CPLS prior to subtotal tumor resection, plus four injections after surgery, is now alive and healthy 22 months after the commencement of treatment, with no detectable serum alpha-fetoprotein (AFP), although the AFP level on admission was 695,000 ng/ml. In the surgical specimens of case 2, there were areas with Lipiodol deposits rich in platinum and replaced by marked fibrosis around necrotic tumor nodules. Intrahepatic arterial injection of CPLS in combination with systemic chemotherapy and surgery should be considered as an effective method for unresectable cases. PMID- 2555661 TI - Summary of notifiable diseases, United States, 1988. PMID- 2555660 TI - Small cell carcinoma of the pancreas in association with a choledochal cyst: immunohistochemical characterization and complete response to combination chemotherapy. AB - Carcinomas arising in choledochal cysts are well recognized; however, small cell carcinomas have not been reported to arise in choledochal cysts. Extrapulmonary small cell carcinomas have been recognized as unique clinicopathologic entities for decades, and have been found arising in numerous sites. Recently there have been several reviews of the subject (Remick et al.: Medicine 66:457-71, 1987; Richardson and Weiland: Seminars in Oncology 9:484-496, 1982; Levenson et al.: Journal of the National Cancer Institute 67:607-612, 1981). Small cell carcinoma of the pancreas has only rarely been described (Reyes and Wang: Cancer 47:2500 2502, 1981; Corrin et al.: Cancer 31:1523-1527, 1973). These tumors must be distinguished from carcinoid tumors and islet cell tumors, which can often lead to histochemical and histologic confusion (Richardson and Weiland: Seminars in Oncology 9:484-496, 1982). Their response to therapy has not been well documented. We report here a small cell carcinoma of the pancreas that arose within a choledochal cyst and that had immunohistochemical characteristics supporting a neuroendocrine origin but that lacked evidence of islet cell differentiation. A complete response to chemotherapy is also described. PMID- 2555662 TI - [Improving early and late results of hepatectomy for HCC]. AB - The present study reports how we indicate safe and beneficial hepatectomy for hepatocellular carcinomas from the experiences of the consecutive 335 patients treated during 1973 and Dec. 1988. The first question was to appreciate the preoperative prediction of a safe limit of hepatectomy. We established a multiple regression formula consisting of several factors such as CT estimated resection rate, ICG retention rate and patient's age at the end of 1980. Since the beginning of 1981, we have been employing the formula as a major indicator guiding treatment option, leading to a significant drop of postoperative, liver failure. The influencing prognostic factors have been determined to be vascular invasion, intrahepatic metastasis, extent of resection regulated by hepatic reserve and tumor size in the decreasing order. From the viewpoint of the biological aspect, the tumors with DNA diploid pattern showed significantly better prognosis than those with non-diploid ones. The current surgical techniques of hepatectomies with en-bloc ligation of Glisson's vessels or with vascular exclusion supported by veno-venous bypass have greatly encouraged the recent surgical results achieving 56% of 4-year survival rate (1985-1988). PMID- 2555663 TI - [Usefulness of endotoxin removal from the septic blood with direct hemoperfusion using PMX-F]. AB - Sepsis and septic shock have been drugs difficult to treat despite many our therapeutic methods. There are many drugs for sepsis, but few can remove or detoxify such sabstances as endotoxin. The polystyrene fiber immobilized polymyxin B (PMX-F) have been invented and it was already confirmed the stability of its bonding. In this paper, mass-productive PMX-F for the clinical application were estimated. IN VITRO STUDIES: PMX-F adsorbed endotoxin (10ng/ml) from the calf serum, and recombinant tumor necrosis factor (80U/ml) from the serine solution. PMX-F maintained an antimicrobial activities against E.coli sufficiently. Those activities of PMX-F showed significant difference between PMX F and carrier fiber. But PMX-F could not adsorb histamine and thromboxane B2. In vivo studies: The treatment of endotoxic mongrel dogs showed early recovery in arterial blood pressure, and ameliorated survival rate significantly. In human study, 77-year old male patients who had obstructive jaundice due to recurrent stomach cancer and biliary tract infection by Klebsiella showed sepsis and DIC. During DHP with PMX-F, the change of blood pressure was remarkable better, and platelet counts decreased, but next morning this count recovered completely and he had no hemorrhagic tendency. We believe clinical application of PMX-F is safe and useful. PMID- 2555664 TI - [Surgery for pleomorphic adenoma of the parotid gland evaluated by long term follow-up]. AB - Fifty six cases with pleomorphic adenoma of the parotid gland were evaluated on local recurrences following surgical treatments which were performed from 1970 to 1983. The surgical treatments for the tumors were 11 extirpations and 45 parotidectomies. The recurrence in each treatment was observed in one (9%) and in none (0%), respectively. The average follow-up period is 11 years (from 5 to 19). The treatment of choice for pleomorphic adenoma is the parotidectomy, of the superficial, deep and/or posterior, lobes. PMID- 2555665 TI - [Surgical treatment for intraductal proliferative lesions]. AB - Intraductal proliferative lesions of the breast including benign, borderline and malignant diseases continue to be one of the most difficult diagnostic and therapeutic problems to the pathologist and surgeon. The optimal management should be done according to the different characteristics of the diseases. However, it is difficult to obtain the precise information of the individual disease before surgery. Selected ductolobular segmentectomy of the breast was performed in 73 patients who had been preoperatively diagnosed as having intraductal papilloma due to the clinical examinations. The specimen obtained from 25 patients were subjected to 3-dimensional (3-D) reconstruction analysis of their duct architectures. As the result, intraductal papilloma was divided into central papilloma and peripheral one. It revealed that none of the nine patients originating in the central duct had carcinoma and 6 of the 16 (37.5%) patients with peripheral papilloma coexisted with carcinoma in the terminal ductal-lobular units (TDLUs). This procedure is consisted of the total excision of affected ductolobular system. The favorable results is obtained in the cosmesis and breast feeding, and there is no recurrent case for the follow up period from 1 to 13 years. PMID- 2555666 TI - [Nuclear DNA analysis of hepatocellular carcinoma]. AB - Retrospective DNA ploidy studies of paraffin-embedded blocks were performed by flow cytometry on 212 surgically resected hepatocellular carcinoma. One hundred and sixty-four of 212 specimens yielded evaluable DNA histograms. Eighty-three cases showed a DNA diploid pattern and 81 a DNA non-diploid pattern. The incidence of non-diploid pattern increased with the tumor size. (2cm; 15.4%, 2-5; 42.3%, 5-10; 70.8% greater than 10; 73.3%). The DNA pattern correlated with the backgrounds, the degree of vascular invasion, intrahepatic metastasis and serum AFP levels. Survival of patients with non-diploid pattern was significantly less than those with diploid patients in any different stage. Of thirty who survived three years with disease free condition, 20 patients showed diploid patterns. Though remaining 10 patients were non-diploid patterns, 9 were high DNA index more than 1.5. It is concluded that flow cytometric DNA ploidy study had prognostic value for patients with surgically resected hepatocellular carcinoma. PMID- 2555667 TI - asgB, a gene required early for developmental signalling, aggregation, and sporulation of Myxococcus xanthus. AB - The asgB genetic locus of Myxococcus xanthus specifies a function which is required early in the developmental pathway leading to aggregation and sporulation in fruiting bodies. The developmental defect of asgB mutants can be compensated by extracellular complementation using either intact wild-type cells or cell-free supernatants conditioned by developing wild-type cells. A Tn5 insertion was isolated closely linked to asgB480 and facilitated the cloning of both the wild-type (asgB+) and the mutant (asgB480) alleles in Escherichia coli plasmid. Tandem duplications of the asgB locus were constructed in M. xanthus; the completely wild-type phenotype of asgB+/asgB480 partial diploids implies that the asgB480 allele is recessive. This finding, along with extracellular complementation by wild-type cells, is consistent with the hypothesis that the asgB+ locus is required to produce a substance with an intercellular signalling function. At least part of the asgB gene was found to lie within a 1.2 kb SmaI DNA fragment. This 1.2 kb fragment, as well as smaller fragments derived from it, were used as DNA probes in RNA/DNA hybrid analyses of transcription in the asgB region. Two small mRNA species were detected, one about 650 bp long, and the other about 500 bp; the two species of mRNAs apparently overlap. Both mRNAs are present in low, but approximately equal amounts, in vegetatively growing cells. This is consistent with the observation that asg mutants display a mutant vegetative phenotype (a change in colony color and spreading behavior) as well as defective development. PMID- 2555668 TI - DNA damage activates transcription and transposition of yeast Ty retrotransposons. AB - A set of genes isolated from Saccharomyces cerevisiae showed increased transcript levels after yeast had been exposed to ultraviolet (UV) light or 4-nitroquinoline 1-oxide (4NQO). Included among these DNA damage responsive (DDR) genes were members of the Ty retrotransposon family of yeast. Northern hybridization analysis indicated that maximal levels of a 5.6 kb transcript encoded by the Ty elements accumulated in cells after 4 to 6 h of exposure to 4NQO. The induced levels of transcripts varied from two- to tenfold for different Ty probes although similar kinetics and dose responses were observed for transcripts hybridizing to the different Ty family members. Pulse labeling experiments suggested that the accumulation of Ty transcripts was due, in part, to an increased rate of Ty message synthesis. Transposition of Ty elements to two target loci encoding distinct alcohol dehydrogenase enzymes, ADH2 and ADH4, was examined in cells exposed to increasing doses of UV light or 4NQO. The frequency of Ty insertion into these genetic regions following DNA damaging treatments increased by as much as 17-fold compared with untreated cells. These results provide direct evidence that transposable elements can be activated by physical and chemical mutagens/carcinogens and that transpositional mutagenesis is induced by these agents in S. cerevisiae. PMID- 2555669 TI - Bacteriophage lambda-mediated transposon mutagenesis of phytopathogenic and epiphytic Erwinia species is strain dependent. AB - Using transformation and conjugal mobilization, plasmids carrying the lamB gene of Escherichia coli were transferred to a range of Erwinia strains. The resultant strains were infected with lambda 467, and kanamycin resistant transductants were screened for various mutant phenotypes including auxotrophy and altered extracellular enzyme activities. Reversion analysis suggested that most mutant phenotypes were due to Tn5 insertion. The applicability of the techniques was highly strain dependent. However a rapid and simple route to mutant isolation was obtained, which could allow the use of other lambda-related genetic techniques in several important species which, to date, have not been genetically manipulated. PMID- 2555670 TI - Characterization and nucleotide sequence of a novel gene fixW upstream of the fixABC operon in Rhizobium leguminosarum. AB - On the Rhizobium leguminosarum PRE sym plasmid, fixABC and a novel gene fixW were identified upstream of the regulatory gene nifA. The molecular masses of FixABC, 29, 44 and 50 kDa respectively, were estimated by polyacrylamide gel electrophoresis (PAGE) and of FixW, 25 kDa, by PAGE and nucleotide sequencing. Hybridization studies using bacteroid mRNA as a probe showed that fixABC is one operon which can be transcribed independently of fixW. Nucleotide sequencing revealed that both fixW and fixA are preceded by a nif consensus promoter. The fixA promoter partly overlaps the 3'-terminal coding region of fixW, indicating that readthrough from fixW into fixA is possible. Two open reading frames, ORF71 and ORF79, precede fixW and form one operon with fixW. ORF71 contains sequences homologous to the fixA promoter and 5'-terminal coding region. One more duplication of fixA sequences was detected, also located within the sym plasmid nif/fix clusters. One duplication of fixW sequences was found. No fixW homologue could be found in other nitrogen fixing organisms except in a number of R. leguminosarum strains. PMID- 2555671 TI - Primary structure of the tms and prs genes of Bacillus subtilis. AB - The nucleotide sequence was determined of a 3211 nucleotide pair EcoRI-PvuII DNA fragment containing the tms and prs genes as well as a part of the ctc gene of Bacillus subtilis. The prs gene encodes phosphoribosylpyrophosphate (PRPP) synthetase, whereas the functioning of the tms and ctc gene products remains to be established. The prs gene contains an open reading frame of 317 codons resulting in a subunit Mr of 34828. An open reading frame comprising the tms gene contained 456 codons resulting in a putative translation product with an Mr of 49,554. Comparison of the deduced B. subtilis PRPP synthetase amino acid sequence with PRPP synthetases from Escherichia coli and rat liver showed extensive similarity. The deduced Tms amino acid sequence was found to be 43% similar to the deduced amino acid sequence of ecourfl, a gene of E. coli with unknown function. PMID- 2555672 TI - Adenosine is required for ethanol-induced heterologous desensitization. AB - Recent evidence suggests that ethanol initially causes an increase in receptor dependent cAMP levels, followed by heterologous desensitization of receptors coupled to GS after chronic exposure. Here we investigated the role of adenosine in mediating these responses. We found that ethanol caused accumulation of extracellular adenosine in NG108-15 and S49 lymphoma cells. This adenosine activated adenosine receptors to increase intracellular cAMP levels. The addition of adenosine deaminase, to degrade accumulated extracellular adenosine, or isobutyl-methylxanthine, an adenosine receptor antagonist, completely blocked ethanol-induced increases in cAMP levels in NG108-15 cells. Chronic exposure of NG108-15 and S49 wild type cells to ethanol resulted in heterologous desensitization of adenosine receptor- and prostaglandin E1 receptor-dependent cAMP signal transduction. Coincubation of NG108-15 and S49 wild type cells with adenosine deaminase and ethanol for 48 hr prevented heterologous desensitization. Moreover, mutant S49 cells, which are unable to transport adenosine, did not accumulate extracellular adenosine after incubation with ethanol and did not develop ethanol-induced heterologous desensitization. Our results suggest that adenosine is an important mediator of both the acute and chronic effects of ethanol on cAMP signal transduction. PMID- 2555673 TI - Regulation of muscarinic receptor subtypes and their responsiveness in rat brain following chronic atropine administration. AB - Chronic administration of l-atropine to rats caused a dose-dependent increase (30%) in the density of muscarinic receptors, as measured with [3H]quinuclidinyl benzilate ([3H]QNB], in cortex (CTX), dorsal hippocampus (DH), and heart but not the corpus striatum. Serum concentrations of l-atropine reached 80 to 160 nM within 6 hr, whereas densities of binding sites for [3H]QNB did not show a significant increase until after the second day of infusion and receptor densities did not reach new steady state levels until after the fourth day of infusion. The density of binding sites for the M1-selective muscarinic receptor antagonist, [3H]pirenzepine ([3H]PZ) was also measured. As noted previously, the density of binding sites for [3H]PZ (defined as M1) was lower than the density of binding sites for [3H]QNB (defined as M1 plus M2). When the densities of binding sites for [3H]QNB and [3H]PZ in CTX plus DH were determined after 14 days of treatment, [3H]QNB binding sites showed a 28% increase, whereas [3H]PZ binding sites did not show any increase. The difference between the densities of binding sites for [3H]QNB and [3H]PZ, an estimate of the density of M2 sites, doubled. The density of binding sites for [3H]QNB appeared to be stable for at least 64 hr after the withdrawal of the drug. The increase in the density of binding sites for [3H]QNB was not reflected in the binding of [3H]oxotremorine-M ([3H]OXO-M), a muscarinic agonist, which was unchanged by l-atropine administration. Because the binding of [3H]OXO-M is sensitive to GTP, this observation suggests that the "induced" receptors may not be coupled to a guanine nucleotide-binding protein. In spite of the fact that there was a doubling of the density of M2 sites, no significant differences in dose-response curves for carbachol-induced inhibition of [3H]cAMP accumulation were observed in slices of CTX plus DH from control and l-atropine-treated rats. Similarity, acetylcholine-stimulated accumulation of [3H]inositol phosphates in slices of CTX plus DH showed no significant differences between the tissues from control and treated rats.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2555674 TI - Ifenprodil is a novel type of N-methyl-D-aspartate receptor antagonist: interaction with polyamines. AB - We have investigated the interactions of polyamines and the N-methyl-D-aspartate (NMDA) receptor antagonist ifenprodil with the binding of [3H]MK801 to the NMDA receptor. Spermine and spermidine but not putrescine substantially increase [3H]MK801 binding to well washed rat brain membranes in the absence or presence of saturating concentrations of glutamate and glycine. Spermine also increased the association and dissociation of [3H]MK801 from its binding site, suggesting that polyamines activate the NMDA receptor in a similar manner to glycine. Ifenprodil inhibited the binding of [3H]MK801 in a biphasic fashion. The high affinity phase of binding (Ki of approximately 15 nM) accounted for 50-60% of total [3H]MK801 binding in the nominal absence of glutamate, glycine, and polyamines or in the presence of 100 microM glutamate. This fraction was reduced to 20% by the addition of 30 microM glycine and could be abolished by the addition of 50 microM spermine. However, ifenprodil apparently did not act by binding to the polyamine recognition site. The low affinity phase (Ki of 20-40 microM) was insensitive to the presence of positive modulators and may represent binding to the Zn2+ regulatory site. Ifenprodil decreased NMDA and glycine induced Ca2+ influx into cultured rat brain neurons. The potency of ifenprodil suggests that spermine may activate NMDA receptors in vivo. These data indicate that ifenprodil may bind to the NMDA receptor in a state-dependent fashion and preferentially stabilize an inactivated form of the channel. PMID- 2555675 TI - Inhibition and stimulation of photoreceptor phosphodiesterases by dipyridamole and M&B 22,948. AB - Few high affinity inhibitors of the photoreceptor phosphodiesterases have been identified. We show here that dipyridamole and M&B 22,948 (Zaprinast), potent inhibitors of the cGMP-binding, cGMP-specific phosphodiesterase (PDE), also inhibit trypsin- or transducin-activated bovine rod and cone photoreceptor phosphodiesterases at submicromolar concentrations. Dixon plots demonstrated that the inhibition of trypsin-activated rod PDE was competitive, with Ki values of 140 nM for M&B 22,948 and 380 nM for dipyridamole. Both of these drugs were much more potent than other PDE inhibitors, including isobutylmethylxanthine (IBMX). These results reinforce the suggestion that the photoreceptor and the cGMP binding, cGMP-specific PDE are closely related. In addition, the high affinity and selectivity of these agents should make them useful for probing the regulation and function of PDE in the photoreceptor. At low substrate concentrations, the effects of these drugs on basal unactivated PDE activity were similar to those seen with trypsin- or transducin-activated PDE. At millimolar substrate concentrations, however, the effects of the drugs were biphasic; PDE activity was stimulated at drug concentrations from 1 to 10 microM and inhibited at higher concentrations. Stimulation was not observed with IBMX. This stimulation of activity apparently was not an allosteric effect caused by direct binding of the dipyridamole and M&B 22,948 to the high affinity noncatalytic cGMP binding sites on the PDEs; whereas no cooperativity of cGMP binding to this site has been demonstrated, the drugs actually stimulated the binding of low concentrations of cGMP to this site. In addition, whereas preincubation with cGMP and cGMP analogs blocked the stimulation exerted by the drugs, they did so only at much higher concentrations than those necessary for saturation of the high affinity noncatalytic cGMP site. Because the stimulation can only be seen at higher substrate levels than are thought to exist in the photoreceptor, only the inhibitory effects of the drugs are likely to be pharmacologically relevant. However, the stimulation exerted by these drugs may point to a hitherto unknown allosteric interaction between the catalytic and regulatory sites on the PDE or to a previously unrecognized regulatory site. PMID- 2555676 TI - Batrachotoxin and alpha-scorpion toxin stabilize the open state of single voltage gated sodium channels. AB - The combined effects of batrachotoxin (BTX) and either scorpion (Leiurus quinquestriatus quinquestriatus) venom (LqqV) or alpha-scorpion toxin (alpha LqqTX) purified from LqqV on single voltage-gated Na channels were studied in planar lipid bilayers. In the presence of BTX, LqqV caused the channels to remain open at membrane potentials at least 50 mV more hyperpolarized than with BTX alone. alpha-LqqTX mimicked the effect of LqqV, suggesting that this toxin is the active component of the venom. LqqV did not significantly alter single-channel conductance, voltage-dependent block by saxitoxin, or voltage-dependent block by Ca2+, indicating that the venom preferentially affects gating rather than ion permeation. The results indicate that a cooperative interaction between alpha LqqTX and BTX strongly favors the open state of the Na channel by causing a large hyperpolarizing shift in the voltage dependence of activation. This effect on activation gating is not predicted from the individual effects of the toxins. PMID- 2555677 TI - Chronic clonazepam administration decreases gamma-aminobutyric acidA receptor function in cultured cortical neurons. AB - Chronic benzodiazepine administration has been reported to decrease gamma aminobutyric acidA (GABAA) receptor function in animals and may alter benzodiazepine binding in neuronal cultures. To assess GABAA receptor function in neuronal cultures exposed to chronic clonazepam, we measured muscimol-stimulated chloride uptake in chick cerebral cortical cultures treated acutely and for 2, 4, and 10 days. Acute clonazepam administration (1 microM) led to an increase in GABA-related chloride uptake at lower doses of muscimol. After chronic clonazepam (1 microM), maximal uptake was markedly decreased at day 10, but maximal uptake was unchanged after 2- and 4-day treatments. Benzodiazepine receptor binding was decreased by approximately 60% after 10 days due to a decrease in receptor number. Decreases in chloride uptake were also observed after 10 days of treatment with 0.1 and 10 microM clonazepam. Concomitant treatment with 0.1 microM Ro15-1788 abrogated the effect of 0.1 microM clonazepam on chloride uptake. Chronic clonazepam treatment (1 microM) did not alter total cellular protein, cellular protein synthesis or degradation or percentage of neuronal cells, as determined morphologically and by [3H]ouabain binding. PMID- 2555679 TI - Changes in the activity of enzymes, participating in glycogen metabolism of alloxan diabetic rats. AB - Alloxan diabetes induced in white rats by intraperitoneal injection of alloxan monohydrate (15 mg/100 g body weight) was used to study changes in the glycogen phosphorylase a and b, phosphoprotein phosphatases and hexokinase activities under insulin deficiency conditions. Among the enzymes studied, an increase in muscle phosphorylase a activity as well as the a/b ratio have been obtained. In diabetic muscle phosphoprotein phosphatases and hexokinase activities were diminished. AMP increased the liver glycogen phosphorylase activity twice in diabetic rats whereas in normal animals the enzyme was less sensitive to this effector. The changes in liver hexokinase activity at diabetes were not connected and correlated with the altered phosphorylase and protein phosphatase activities. The logical chain of probable molecular events taking place in muscle glycogen metabolism under the conditions of insulin deficiency is offered. PMID- 2555678 TI - Effect of calcium overload on the phosphoinositide breakdown in the rat left ventricular papillary muscle. AB - We investigated the effect of Ca2+ overload on the phospholipase C-catalyzed hydrolysis of phosphoinositides in the rat left ventricular papillary muscle. Ca2+ overload on the papillary muscle was induced by treatment with 0.3 mM ouabain in Ca2+-containing medium following either Ca2+-containing or Ca2+-free superfusion. The phosphoinositide breakdown was evaluated by determining accumulations of [3H]inositol phosphates ([3H]IPs) in the tissues prelabeled with [3H]inositol. Ca2+ repletion following Ca2+-free superfusion resulted in a rapid but small increase in resting tension that was not followed by contracture, nor was it associated with a significant increase in [3H]IPs accumulations. Treatment with ouabain following Ca2+-containing superfusion increased resting tension after a lag period of several minutes and produced contracture associated with an increase in [3H]IPs accumulations. The ouabain induced increases in resting tension, and accumulations of [3H]IPs were significantly potentiated by prior Ca2+-free superfusion instead of Ca2+-containing superfusion. There was a significant positive correlation between increases in resting tension and the phosphoinositide breakdown. The increased resting tension and the accumulations of [3H]IPs were not antagonized by treatments with prazosin plus atropine or indomethacin, but were abolished by superfusion with Ca2+-free buffer solution. Although the enhanced phospholipase C-catalyzed hydrolysis of phosphoinositides appears to be a consequence rather than a cause of increased intracellular Ca2+, such a biochemical change may provoke a positive feedback mechanism to develop the muscle contracture through the putative intracellular messenger action of inositol triphosphate and diacylglycerol. PMID- 2555681 TI - [Paths in graphs and selection of oligonucleotide linkers]. AB - The problem of search of an universal linker with minimal length containing all restriction endonuclease recognition sites is considered. We reduce this problem to the search of Eiler's and Hamilton's paths in graph. The use of the discrete optimization methods allows to construct the linker with the length closed to the minimum. PMID- 2555680 TI - Metabolic adaptation of the renal carbohydrate metabolism. III. Effects of high protein diet on the gluconeogenic and glycolytic fluxes in the proximal and distal renal tubules. AB - The adaptive response of renal metabolism of glucose was studied in isolated rat proximal and distal renal tubules after a high protein-low carbohydrate diet administration. This nutritional situation significantly stimulated the gluconeogenic activity in the renal proximal tubules (about 1.5 fold at 48 hours) due, in part, to a marked increase in the fructose 1,6-bisphosphatase (FBPase) and phosphoenolpyruvate carboxykinase (PEPCK) activities. In this tubular fragment, FBPase activity increased only at subsaturating fructose 1,6 bisphosphate concentration (30% at 48 hours) which involved a significant decrease in the Km (31%) for its substrate without changes in the Vmax. This enzymatic behaviour is probably related to modifications in the activity of the enzyme already present in the renal cells. Proximal PEPCK activity progressively increased at all substrate concentrations (almost 2 fold at 48h of high protein diet) which brought about changes in Vmax without changes in Kim. These changes are in agreement with variations in the cellular concentration of the enzyme. Neither gluconeogenesis nor the gluconeogenic enzymes changed in the distal fractions of the renal tubules. On the other hand, a high protein diet did not apparently modify the glycolytic ability in any fragment of the nephron, although a significant increase in the phosphofructokinase (PFK) and pyruvate kinase (PK) activities was found in the distal renal tubules. This short term regulation involved a significant decrease from 24 hours in the Km value of distal PFK (almost 40%) without changes in Vmax. The kinetic behaviour of distal PK was mixed. In the first 24h after high protein diet a significant decrease in the Km for phosphoenolpyruvate was found (30%) without variation in the Vmax, however during the second 24 hours the activity of this glycolytic enzyme increased significantly (almost 1.3 fold) without modifications in its Km value. On the contrary, this nutritional state did not modify the kinetic behaviour of any glycolytic enzyme in the proximal regions of the renal tubules. PMID- 2555682 TI - [Quantum-chemical study of the relative stability of nucleic acid bases protonation stability]. AB - The influence of protonation on the relative stability of matched and mismatched nucleic base pairs was considered. Protonation of a base has a significant effect on the bond energy and, in some cases, on the geometry of pairs. It was shown that protonation of guanine, adenine and uracil lead to an increased relative stability of matched pairs and, consequently, to decreased probability of mutations. Protonation of cytosine reduced the difference in energy formation of GC and AC complexes. PMID- 2555683 TI - [Bases for multiplication and genetic variability of retroviruses. The murine leukemia virus (MuLV) and AIDS virus (HIV-1)]. AB - The review describes the replication and genetic variability of retroviruses. The steps of the replication cycle are detailed for murine leukemia (MuLV) and AIDS (HIV-1) viruses. PMID- 2555684 TI - Indirect binding of human T-cell leukemia virus type I tax1 to a responsive element in the viral long terminal repeat. AB - Several laboratories have demonstrated that tandem copies of the human T-cell leukemia virus type I 21-base-pair (bp) repeat cloned upstream of either a homologous or heterologous promoter increase transcription in the presence of tax1 protein. In this report, we provide evidence for a second tax1-responsive sequence in the viral long terminal repeat. Analysis of human T-cell leukemia virus type I promoter deletion mutants and plasmids containing cloned oligonucleotide motifs demonstrated that this 47-bp sequence, located between 117 and -163, confers responsiveness to tax1. We further demonstrated that proteins present in HeLa nuclear extracts bind specifically to this tax1 responsive sequence. Mutants that affected in vivo activity also decreased in vitro binding. Using an in vitro binding assay, we demonstrated that tax1 interacts indirectly with the 47-bp sequence, most likely through protein-protein interaction. Thus, while tax1 does not bind directly to DNA to enhance transcription, it may influence sequence-specific responses by interacting with the primary DNA-protein complex. PMID- 2555685 TI - Expression and organization of BP74, a cyclic AMP-regulated gene expressed during Dictyostelium discoideum development. AB - We have characterized a cDNA and the corresponding gene for a cyclic AMP inducible gene expressed during Dictyostelium development. This gene, BP74, was found to be first expressed about the time of aggregate formation, approximately 6 h after starvation. Accumulation of BP74 mRNA did not occur in Dictyostelium cells that had been starved in fast-shaken suspension cultures but was induced in similar cultures to which cyclic AMP pulses had been added. The BP74 cDNA and gene were characterized by DNA sequence analysis and transcriptional mapping. When the BP74 promoter region was fused with a chloramphenicol acetyltransferase reporter gene and reintroduced into Dictyostelium cells, the transfected chloramphenicol acetyltransferase gene displayed the same developmentally regulated pattern of expression as did the endogenous BP74 gene, suggesting that all of the cis-acting elements required for regulated expression were carried by a 2-kilobase cloned genomic fragment. On the basis of sequence analysis, the gene appeared to encode a protein containing a 20-residue hydrophobic sequence at the amino-terminal end and 26 copies of a 20-amino-acid repeat. PMID- 2555686 TI - Multiple forms of poly(A) polymerases purified from HeLa cells function in specific mRNA 3'-end formation. AB - Poly(A) polymerases (PAPs) from HeLa cell cytoplasmic and nuclear fractions were extensively purified by using a combination of fast protein liquid chromatography and standard chromatographic methods. Several forms of the enzyme were identified, two from the nuclear fraction (NE PAPs I and II) and one from the cytoplasmic fraction (S100 PAP). NE PAP I had chromatographic properties similar to those of S100 PAP, and both enzymes displayed higher activities in the presence of Mn2+ than in the presence of Mg2+, whereas NE PAP II was chromatographically distinct and had approximately equal levels of activity in the presence of Mn2+ and Mg2+. Each of the enzymes, when mixed with other nuclear fractions containing cleavage or specificity factors, was able to reconstitute efficient cleavage and polyadenylation of pre-mRNAs containing an AAUAAA sequence element. The PAPs alone, however, showed no preference for precursors containing an intact AAUAAA sequence over a mutated one, providing further evidence that the PAPs have no intrinsic ability to recognize poly(A) addition sites. Two additional properties of the three enzymes suggest that they are related: sedimentation in glycerol density gradients indicated that the native size of each enzyme is approximately 50 to 60 kilodaltons, and antibodies against a rat hepatoma PAP inhibited the ability of each enzyme to function in AAUAAA-dependent polyadenylation. PMID- 2555687 TI - Multiple sequence elements of a single functional class are required for cyclic AMP responsiveness of the mouse c-fos promoter. AB - Agents that elevate the intracellular concentration of cyclic AMP (cAMP) rapidly and transiently induce expression of the c-fos proto-oncogene in BALB/c 3T3 cells. We show that the mouse c-fos promoter-enhancer region contains multiple elements that contribute to cAMP responsiveness of the promoter in transient expression assays. The most potent element was found to correspond to a previously mapped basal promoter element and protein-binding site located 65 base pairs upstream of the transcriptional initiation site. This element and two less potent sites contained a match to the cAMP response element (CRE) core sequence defined in several mammalian genes. The relative potencies of these elements corresponded with their relative affinities for cellular factors that bound to the CRE in vitro. Mutation of all three elements failed to abolish completely cAMP responsiveness of the c-fos promoter in the transient expression assay. However, we present evidence that this residual responsiveness may have been due to sequences present in vector DNA. Finally, we show, by using a new microinjection competition assay, that a double-stranded oligonucleotide carrying the major c-fos CRE is sufficient to block induction of the endogenous c-fos gene by cAMP. Therefore, induction of the endogenous gene requires positively acting cellular factors that interact with a single functional class of regulatory sites in the c-fos gene. Unrelated regulatory elements, such as the serum response element and putative AP-2 sites, are not by themselves sufficient to mediate the cAMP response. PMID- 2555688 TI - Possible involvement of normal p21 H-ras in the insulin/insulinlike growth factor 1 signal transduction pathway. AB - Expression of a mutant H-ras gene confers a transformed phenotype to rat-1 fibroblasts which is basically independent of exogenous growth factors (GFs). Rat 1 cells induced to express high levels of the normal H-ras gene were also found to display a transformed phenotype. In contrast to cells expressing mutant H-ras, these cells were dependent on GFs. We used this difference in GF dependence to analyze a possible involvement of exogenous GFs in H-ras function. Compared with untransformed rat-1 cells, cells overexpressing normal H-ras displayed an elevated response toward insulinlike growth factor 1 (IGF-1), insulin, and bombesin and an increased sensitivity toward phosphatidic acids. It was found that 8-bromo-cyclic AMP inhibited the responses to all GFs in rat-1 cells but had no effect on mutant-H-ras-transformed cells. In cells overexpressing normal H ras, 8-bromo-cyclic AMP inhibited the responses to all GFs except those to insulin and IGF-1. This implies that overexpression of normal H-ras in the presence of insulin/IGF-1 is functionally similar to the expression of mutant H ras, since mutant H-ras can circumvent this block by itself. These and other results strongly suggest a functional linkage between insulin/IGF-1 and normal p21 H-ras. PMID- 2555689 TI - Transactivation of gene expression by nuclear and cytoplasmic rel proteins. AB - Transcriptional activation of gene expression by oncogenic proteins can lead to cellular transformation. It has recently been demonstrated that the protein encoded by the v-rel oncogene from reticuloendotheliosis virus strain T can transactivate gene expression from certain promoters in a cell-specific manner. We have examined the cytological location, transforming properties, and transactivation properties of proteins encoded by chimeric turkey v-rel/chicken c rel genes. We found that whereas the v-rel protein was nuclear in both chicken embryo and rat fibroblasts, the presence of the C terminus of the c-rel protein inhibited nuclear localization of the rel protein in these fibroblasts. Cytoplasmic rel proteins containing C-terminal c-rel sequences transactivated gene expression from the polyomavirus late promoter as efficiently as did similar rel proteins located in the nucleus. These results indicate that the cellular location of rel proteins is not important for transactivation of gene expression and suggest that transactivation by rel proteins is indirect, perhaps by affecting an intracellular signal transduction pathway that eventually results in the alteration of gene expression. The transforming properties of the rel protein were unaltered by the presence of the c-rel C terminus, but, as previously reported for turkey c-rel sequences, substitution of chicken c-rel sequences for internal v-rel sequences reduced the transforming activity of the rel protein and eliminated the immortalization ability. However, all of the chimeric v/c-rel proteins were able to transactivate gene expression, indicating that transactivation does not correlate with transformation. These results suggest that transactivation may be necessary but is not sufficient for transformation by rel proteins. PMID- 2555690 TI - A small nuclear RNA, U5, can transform cells in vitro. AB - Low-molecular-weight RNA exhibiting transforming potential was identified in chemically induced lymphoma cells by the transformation of mink lung cells after transfection. The RNA was sequenced by the direct chemical method and was shown to be a small nuclear RNA, U5. The transforming potential of the RNA was further studied in quantitative transformation assays using 3Y1, a rat fibroblastic cell line. Transformed foci appeared with a latency of 3 to 4 weeks after transfection. U5-transformed 3Y1 cells frequently carried an amplified c-myc oncogene. In addition, U5 induced chromosome aberrations in transfected cells, indicating that the RNA acts as a clastogen. Transforming and clastogenic potentials were specifically inactivated when U5 was incubated with RNase H in the presence of a complementary oligonucleotide. We discuss a possible mechanism of U5-induced cell transformation. PMID- 2555691 TI - cis-acting elements and a trans-acting factor affecting alternative splicing of adenovirus L1 transcripts. AB - Expression of the L1 region of adenovirus is temporally regulated by alternative splicing to yield two major RNAs encoding the 52- to 55-kilodalton (52-55K) and IIIa polypeptides. The distal acceptor site (IIIa) is utilized only during the late phase of infection, whereas the proximal site (52-55K) is used at both early and late times. Several parameters that might affect this alternative splicing were tested by using expression vectors carrying the L1 region or mutated versions of it. In the absence of a virus-encoded or -induced factor(s), only the 52-55K acceptor was used. Decreasing the distance between the donor and the IIIa acceptor had no effect. Removal of the 52-55K acceptor induced IIIa splicing slightly, implying competition between the two acceptors. Fusion of the IIIa exon to the 52-55K intron greatly enhanced splicing of the IIIa junction, suggesting that the IIIa exon does not contain sequences that inhibit splicing. Thus, the lack of splicing to the IIIa acceptor in the absence of a virus-encoded or induced factor(s) is probably due to the absence of a favorable sequence and/or the presence of a negative element 5' of the IIIa splice junction, or both. The presence of several adenovirus gene products, including VA RNAs, the E2A DNA binding protein, and the products of E1A and E1B genes, did not facilitate use of the IIIa acceptor. In contrast, the simian virus 40 early proteins, probably large T antigen, induced IIIa splicing. This result, together with those of earlier studies, suggest that T antigen plays a role in modulation of alternative RNA splicing. PMID- 2555692 TI - Distinguishable promoter elements are involved in transcriptional activation by E1a and cyclic AMP. AB - The sequence motif CGTCA is critical for binding of a group of cellular transcription factors (ATF, CREB, E4F, and EivF) and for activation of certain E1a-inducible and cyclic AMP (cAMP)-inducible promoters. We have tested different promoter elements containing the CGTCA motif (referred to here as ATF-binding sites) for the ability to function as E1a or cAMP response elements. The adenovirus E4 promoter and the cellular vasoactive intestinal peptide (VIP) promoter responded differently to E1a and cAMP, demonstrating that the activating potential of ATF-binding sites within these promoters is not equivalent. While particular ATF-binding sites were sufficient for the activity of both the E4 (E1a inducibility) and VIP (cAMP inducibility) enhancers, these two enhancers had contrasting effects on E1a- and cAMP-inducible transcription. Thus, the relationship between E1a- and cAMP-inducible transcription is not simply explained by the action of these two inducers through the same promoter elements. PMID- 2555693 TI - An intronic 10-base-pair deletion in a class II A beta gene affects RNA processing. AB - Several biologically important examples of posttranscriptionally regulated genes have recently been described (T. Gerster, D. Picard, and W. Schaffner, Cell 45:45 52, 1986; R. Reeves, T.S. Elton, M.S. Nissen, D. Lehn, and K.R. Johnson, Proc. Natl. Acad. Sci. USA 84:6531-6535, 1987; H.A. Young, L. Varesio, and P. Hwu, Mol. Cell. Biol. 6:2253-2256, 1986). Little is known, however, regarding sequences that mediate posttranscriptional RNA stability. Characterization in our laboratory of a mutant murine B lymphoma, M12.C3, revealed a posttranscriptional defect affecting the synthesis of a major histocompatibility complex class II gene (A beta d) whose product normally controls both the specificity and magnitude of the immune response. Molecular studies revealed that the mutation responsible for diminished A beta d gene expression was an intronic deletion of 10 base pairs (bp) located 99 bp 5' of the third exon. This deletion lies in a region not known to be critical for accurate and efficient splicing. Furthermore, sequence analysis of amplified A beta-specific cDNA demonstrated that the small number of A beta d transcripts produced in the mutant cells was correctly spliced. It appears that the mechanism by which this intronic 10-bp deletion acts to decrease RNA stability is unlikely to be at the level of RNA splicing. PMID- 2555694 TI - Fidelity of DNA polymerase I and the DNA polymerase I-DNA primase complex from Saccharomyces cerevisiae. AB - We have determined the fidelity of DNA synthesis by DNA polymerase I (yPol I) from Saccharomyces cerevisiae. To determine whether subunits other than the polymerase catalytic subunit influence fidelity, we measured the accuracy of yPol I purified by conventional procedures, which yields DNA polymerase with a partially proteolyzed catalytic subunit and no associated primase activity, and that of yPol I purified by immunoaffinity chromatography, which yields polymerase having a single high-molecular-weight species of the catalytic subunit, as well as three additional polypeptides and DNA primase activity. In assays that score polymerase errors within the lacZ alpha-complementation gene in M13mp2 DNA, yPol I and the yPol I-primase complex produced single-base substitutions, single-base frameshifts, and larger deletions. For specific errors and template positions, the two forms of polymerase exhibited differences in fidelity that could be as large as 10-fold. Nevertheless, results for the overall error frequency and the spectrum of errors suggest that the yPol I-DNA primase complex is not highly accurate and that, just as for the polymerase alone, its fidelity is not sufficient to account for a low spontaneous mutation rate in vivo. The specificity data also suggest models to explain -1 base frameshifts in nonrepeated sequences and certain complex deletions by a direct repeat mechanism involving aberrant loop-back synthesis. PMID- 2555695 TI - Insertional activation of N-myc by endogenous Moloney-like murine retrovirus sequences in macrophage cell lines derived from myeloma cell line-macrophage hybrids. AB - Hybrids formed from a myeloma cell line, NS1, and macrophages initially show myeloma properties but later, after loss of the parental macrophage genome and consequent loss of myeloma characteristics, express macrophage properties. Molecular studies demonstrated that macrophage properties in the hybridomas originate from the NS1 parental cells (M. Setoguchi, S. Yoshida, Y. Higuchi, S. Akizuki, and S. Yamamoto, Somatic Cell Mol. Genet. 14:427-438, 1988). In such hybrids, N-myc was activated by insertion of endogenous Moloney-like retrovirus sequences into mouse N-myc exon 3 when the hybrids gained macrophage properties. Interestingly, expression of N-myc took place in all aged hybrids. These results suggest that such unique insertional mutagenesis occurs in a regionally specific manner and that expression of N-myc may play a role in hematopoietic lineage conversion. PMID- 2555696 TI - Beta-globin enhancers target expression of a heterologous gene to erythroid tissues of transgenic mice. AB - To examine the role of human beta-globin enhancers in tissue-specific and developmental regulation, a hybrid beta-globin-simian virus 40 gene was analyzed in transgenic mice. A beta-globin DNA fragment containing two previously defined enhancers stimulated transcription from the simian virus 40 promoter in a tissue- and stage-specific pattern similar to that of the normal beta-globin gene. These results help to define the functions of beta-globin regulatory elements and suggest an approach for targeted expression of heterologous genes in erythroid cells in vivo. PMID- 2555697 TI - Identification of an upstream activation sequence and other cis-acting elements required for transcription of COX6 from Saccharomyces cerevisiae. AB - Transcription of Saccharomyces cerevisiae COX6, the nuclear gene for subunit VI of cytochrome c oxidase, is activated in heme-proficient cells, requires the HAP2 gene, and is subject to glucose repression. In this study, by deletion mutagenesis of the COX6 promoter, we identified two regions that are important for transcription. The first was an upstream activation site, UAS6. It was found to be contained within an 84-base-pair (bp) sequence, between bp -256 and -340 of the COX6 translational initiation codon, and to contain sequences required for activation by heme and HAP2 and for release from glucose repression. When located upstream of a CYC1-lacZ fusion gene, deleted for both of its UASs, this segment functioned as a UAS element. Although UAS6 could promote expression in either orientation, it showed a marked orientation dependence in its response to HAP2 and the carbon source. The second region lay between bp -255 and -91. It contained two of the three major 5' termini of COX6 mRNAs and a putative TATA box. Deletion analysis of this region demonstrated that the putative TATA box is not required for transcription and that this region is separable into two redundant domains. PMID- 2555698 TI - Alpha interferon and gamma interferon stimulate transcription of a single gene through different signal transduction pathways. AB - Interferons (IFNs) play a key role in the defense against virus infection and the regulation of cell growth and differentiation, in part through changes in specific gene transcription in target cells. We describe several differences between the signal transduction events that result in transcriptional activation of the human gene coding for a guanylate-binding protein (GBP) by alpha interferon (IFN-alpha) and gamma interferon (IFN-gamma). Activation by IFN-alpha was rapid, transient, and cycloheximide resistant. Activation by IFN-gamma was slower, sustained, and delayed by cycloheximide. IFN-gamma led to the formation of a stable intracellular signal which led to continued GBP transcription even if the ligand was withdrawn, whereas IFN-alpha-induced GBP transcription decayed rapidly if IFN-alpha was withdrawn. Perturbations of signaling pathways involving classical second messengers (cyclic AMP, Ca2+, protein kinase C) did not induce GBP transcription. However, various kinase inhibitors blocked the transcriptional response to IFN-gamma but not IFN-alpha, suggesting that a specific and possibly novel kinase is involved in gene activation by IFN-gamma. PMID- 2555699 TI - Lymphoid and mesenchymal tumors in transgenic mice expressing the v-fps protein tyrosine kinase. AB - src, abl, and fps/fes are prototypes for a family of genes encoding nonreceptor protein-tyrosine kinases. The oncogenic potential of the v-fps protein-tyrosine kinase was investigated by introduction of the gag-fps coding sequence of Fujinami sarcoma virus into the mouse germ line. Transgenic mice with v-fps under the transcriptional control of a 5' human beta-globin promoter (GF) or with both 5' and 3' beta-globin regulatory sequences (GEF) were viable. Unexpectedly, both GF and GEF transgenes were expressed in a wide variety of tissues and induced a spectrum of benign and malignant tumors. These tumors, which included lymphomas, thymomas, fibrosarcomas, angiosarcomas, hemangiomas, and neurofibrosarcomas, developed with various frequencies after latent periods of 2 to 12 months. The majority of lymphoid neoplasms appeared to be of T-cell origin and were monoclonal, as judged by rearrangements of the T-cell receptor beta or immunoglobulin genes. Some tissues that expressed the v-fps oncogene, such as heart, brain, lung, and testes, developed no malignant tumors. The v-fps protein tyrosine kinase therefore has a broad but not unrestricted range of oncogenic activity in cells of lymphoid and mesenchymal origin. The incomplete penetrance of the neoplastic phenotype and the monoclonality of lymphoid tumors suggest that tumor formation in v-fps mice requires genetic or epigenetic events in addition to expression of the P130gag-fps protein-tyrosine kinase. PMID- 2555701 TI - Genetic evidence that acute morphologic transformation, induction of cellular DNA synthesis, and focus formation are mediated by a single activity of the bovine papillomavirus E5 protein. AB - The bovine papillomavirus (BPV) type 1 E5 gene encodes a 44-amino-acid protein that can stably transform cultured rodent cells when expressed in the absence of all other viral genes. We have previously constructed a BPV-simian virus 40 recombinant virus (Pava-1) which efficiently expresses the BPV type 1 E5 gene in infected cells (J. Settleman and D. DiMaio, Proc. Natl. Acad. Sci. USA 85:9007 9011, 1988). Within 48 h of Pava-1 infection, the vast majority of mouse C127 cells underwent a dramatic morphologic transformation which was accompanied by cell proliferation. Infection of C127 cells made quiescent by contact inhibition and serum starvation caused a great induction of cellular DNA synthesis. These morphologic and mitogenic responses were proportional to the virus multiplicity of infection. Mutational analysis indicated that the E5 gene is both necessary and sufficient for these activities. Analysis of a variety of E5 missense mutants revealed a strong correlation between their phenotypes in the acute transformation assays following infection and in the stable focus-forming assay following transfection. Most of the defective mutants expressed normal levels of E5 protein following infection, indicating that their defective phenotypes are not due to the synthesis of an unstable protein. The failure to genetically resolve these E5 activities suggests that the ability of the E5 protein to cause acute morphologic transformation and reentry into the cell cycle may be intimately related to its ability to cause stable cell transformation and that these functions are probably mediated by a single biochemical activity of the E5 protein. PMID- 2555700 TI - Properties of the DNA-binding domain of the simian virus 40 large T antigen. AB - T antigen (Tag) from simian virus 40 binds specifically to two distinct sites in the viral origin of replication and to single-stranded DNA. Analysis of the protein domain responsible for these activities revealed the following. (i) The C terminal boundary of the origin-specific and single-strand-specific DNA-binding domain is at or near amino acid 246; furthermore, the maximum of these DNA binding activities coincides with a narrow C-terminal boundary, spanning 4 amino acids (246 to 249) and declines sharply in proteins with C termini which differ by a few (4 to 10) amino acids; (ii) a polypeptide spanning residues 132 to 246 of Tag is an independent domain responsible for origin-specific DNA binding and presumably for single-stranded DNA binding; and (iii) a comparison of identical N terminal fragments of Tag purified from mammalian and bacterial cells revealed differential specificity and levels of activity between the two sources of protein. A role for posttranslational modification (phosphorylation) in controlling the DNA-binding activity of Tag is discussed. PMID- 2555702 TI - Tissue-specific lability and expression of avian leukosis virus long terminal repeat enhancer-binding proteins. AB - Avian leukosis virus (ALV) induces bursal lymphomas in chickens, after proviral integration next to the cellular myc proto-oncogene, and subsequent c-myc hyperexpression. Our previous work suggested that labile or short-lived cellular proteins interact with the viral long terminal repeat (LTR) enhancer, and binding of these proteins appeared to be essential for high rates of LTR-enhanced transcription (A. Ruddell, M. Linial, W. Schubach, and M. Groudine, J. Virol. 62:2728-2735, 1988). This lability is specific for B-lymphoid cell types, since T cells and fibroblasts show stable high rates of LTR-enhanced transcription and stable LTR-binding activity. Moreover, the lability of these proteins may be important in determining susceptibility to bursal lymphoma. In this study, we separated and characterized the labile and stable LTR-binding proteins and examined their lability and expression in different cell types. Gel shift and DNase I footprinting analyses indicated that at least five proteins interact with the 140-base-pair LTR enhancer region. These proteins were distinct by several criteria, including lability or stability after inhibition of protein synthesis, resistance to heat denaturation, chromatographic behavior, and expression in different cell types. Two binding proteins were present in many cell types and were specifically labile in B cells. A third binding protein showed hematopoietic cell-type-specific expression and was also labile in B cells. These findings indicate that there is tissue-specific modulation of the lability and expression of ALV LTR-binding proteins, which may be important for regulation of LTR transcription enhancement and ALV bursal lymphomagenesis. PMID- 2555703 TI - Tyrosine kinase oncogenes abrogate interleukin-3 dependence of murine myeloid cells through signaling pathways involving c-myc: conditional regulation of c-myc transcription by temperature-sensitive v-abl. AB - Retroviral expression vectors carrying the tyrosine kinase oncogenes abl, fms, src, and trk abrogate the requirements of murine myeloid FDC-P1 cells for interleukin-3 (IL-3). Factor-independent clones constitutively express c-myc in the absence of IL-3, whereas in parental cultures c-myc transcription requires the presence of the ligand. To directly test the effect of a tyrosine kinase oncogene on c-myc expression, retroviral constructs containing three different temperature-sensitive mutants of v-abl were introduced into myeloid IL-3 dependent FDC-P1 and 32D cells. At the permissive temperature, clones expressing temperature-sensitive abl behaved like wild-type abl-containing cells in their growth properties and expressed c-myc constitutively. Temperature shift experiments demonstrated that both IL-3 abrogation and the regulation of c-myc expression correlated with the presence of functional v-abl. Induction of c-myc expression by reactivation of temperature-sensitive v-abl mimicked c-myc induction by IL-3 in that it did not require protein synthesis and occurred at the level of transcription, with effects on both initiation and a transcription elongation block. However, v-abl-regulated FDC-P1 cell growth differed from IL-3 regulated growth in that c-fos and junB, which are normally induced by IL-3, were not induced by activation of v-abl. PMID- 2555705 TI - Metal-specific posttranscriptional control of human metallothionein genes. AB - During the initial 4 h of treatment, copper and zinc similarly activated the rates of transcription and mRNA accumulation from the two human metallothionein (MT) genes, viz., MTI-G and MTII-A, in the hepatoblastoma cell line HepG2. The levels of copper-induced MT mRNAs remained at a plateau for up to 15 h. In contrast, the levels of zinc-induced MT mRNAs gradually declined after about 4 h, despite substantial transcription. The decrease in the zinc-induced MT mRNA half life is probably due to a posttranscriptional event(s). PMID- 2555704 TI - Definition of an Ets1 protein domain required for nuclear localization in cells and DNA-binding activity in vitro. AB - Ets1 and Ets2 are nuclear phosphoproteins which bind to DNA in vitro and share two domains of strong identity. Deletion analyses of each of these conserved regions in Ets1 demonstrated that integrity of the carboxy-terminal domain, also conserved in the more distantly related elk and erg gene products, is essential for both nuclear targeting and DNA-binding activity in vitro. PMID- 2555707 TI - [Nephrologic aspects of tumor diseases in children]. AB - Renal function disturbances in malignant diseases have various origins. Primary tumorous renal infiltrates and secondary renal function impairment via the tumor lysis syndrome are the most frequent causes. Cytostatic drugs are frequently accompanied by serious nephrotoxic side effects, which can be both acute and chronic. The diagnostic methodology for renal function control is simple and is normally available in any laboratory. The prophylactic measures to prevent serious complications are based on a rigid fluid intake, alkalinization of the urine and careful management of the cytoreductive phase. In addition to acute nephrotoxicity, chronic sequelae of renal disturbances may be an important aspect in future long-time studies. PMID- 2555708 TI - Oto-palato-digital syndrome in an Iranian infant. AB - A male infant is presented with wide fontanels, micrognathia, mid-face hypoplasia, hypertelorism, broad nasal root, down-slanting palpebral fissures, small thorax, funnel chest, short wide toes, camptodactyly and cutaneous syndactyly of fingers and toes, dysplastic bones with thin wavy ribs and bowed femore, cryptorchidism, and hypospadias grade I. The mother of this infant showed some signs of the same condition, including hypertelorism, micrognathia, small nose with depressed bridge, flat mid-face, impacted teeth and small chest. This case shows many similarities to oto-palatal-digital syndrome types I and II. PMID- 2555706 TI - Location of signal sequences for membrane insertion of the Na+,K+-ATPase alpha subunit. AB - To study the membrane insertion of the Na+,K+-ATPase (EC 3.6.1.37) alpha subunit with six to eight transmembrane segments, mRNAs encoding the entire alpha subunit and its four different domains were prepared and translated in rabbit reticulocyte lysate with rough microsomal membranes. On the basis of the resistance of the membrane-inserted products to alkali extraction and the failure to insert the translation products into N-ethylmaleimide-treated membranes, it is suggested that at least two signal sequences are contained within the four transmembrane segments from the amino terminus of the alpha subunit. PMID- 2555709 TI - Effect of DNOC, Ferbam and Imidan exposure on mouse sperm morphology. AB - DNOC, Ferbam and Imidan were tested in (C3H X C57BL/6) F1 mice to assess their potential testicular toxicity. Chemicals were administered i.p. and per os at different doses for 5 consecutive days. After 35 days the testicular was toxicity was evaluated by measuring the testicular weights, the sperm counts and the percentage of abnormal sperm. DNOC and Imidan failed to induce teratospermia in mice treated by both routes of administration. Conversely Ferbam induced a statistically significant increase in teratospermia only following per os administration to mice at a dose of 1000 mg/kg b.w./day. These data indicate that per os administration of Ferbam succeeded in producing active metabolites able to interfere with the differentiation process of spermatogenic cells. PMID- 2555710 TI - Two proteins of 220 kD and 230 kD bind to UV-damaged SV40 minichromosomes in irradiated monkey kidney cells. AB - We exposed SV40-infected monkey kidney cells to 0, 30 or 150 J/m2 of UV radiation, and isolated viral minichromosomes at various times after irradiation. Analysis of minichromosome-associated proteins by SDS-PAGE revealed the presence of two proteins, of 220 kD and 230 kD associated with minichromosomes from irradiated cells, but not from unirradiated controls. The larger protein was the less abundant, and was most evident in preparations from more heavily irradiated cells. Neither protein was associated with minichromosomes isolated 30 min after irradiation, but were apparent in minichromosome preparations isolated 1-4 h after UV treatment. PMID- 2555711 TI - G/U lesions are efficiently corrected to G/C in SV40 DNA. AB - Cytosine spontaneously deaminates to form uracil, generating G/U pairs in DNA. We studied the repair of these lesions by introducing specific G/U pairs into the genome of SV40 and determining the fate of the mispaired bases in Simian cells. Analysis of 135 plaques obtained after transfection of the modified viral DNA indicates that G/U lesions were repaired to G/C in every case. This result indicates that G/U lesions are corrected with greater efficiency and specificity than any combination of DNA base/base mispairs, in transfected SV40 DNA. PMID- 2555712 TI - Antifungal activity of diethyldithiocarbamate. AB - Sodium diethyldithiocarbamate (DTC) was evaluated for its ability to combat four different species of fungi in vitro. Using a microtiter-broth-dilution method we were able to demonstrate an antifungal activity against Candida albicans, Cryptococcus neoformans, Aspergillus fumigatus and Mucor mucedo in doses achievable by intravenous administration in man. PMID- 2555713 TI - 3,4-Diaminopyridine in the treatment of Lambert-Eaton myasthenic syndrome. AB - Lambert-Eaton myasthenic syndrome is characterized by muscle weakness, hyporeflexia, and autonomic dysfunction, which result from impaired release of acetylcholine from cholinergic nerve terminals. It is frequently associated with cancer, it is autoimmune-mediated, and treatment has been unsatisfactory. 3,4 Diaminopyridine enhances the release of acetylcholine. In this prospective, double-blind, placebo-controlled crossover study of 12 patients with Lambert Eaton myasthenic syndrome (7 of whom had cancer), 3,4-diaminopyridine in doses up to 100 mg per day was effective in treating both the motor and the autonomic deficits of the syndrome. Muscle strength increased from an average of 70 percent of normal to 81 percent of normal in the upper extremities, and from 45 to 65 percent of normal in the lower extremities. The amplitudes of compound-muscle action potentials nearly doubled, increasing from an average of 2.9 mV to 5.0 mV in the arm and from 1.6 mV to 3.1 mV in the leg. Autonomic symptoms were relieved. One patient had a seizure after 10 months of treatment, but other side effects from the drug were minimal and dose-related. We conclude that 3,4 diaminopyridine, either alone or in conjunction with other therapies, may be useful in the treatment of Lambert-Eaton myasthenic syndrome. PMID- 2555714 TI - Cancer-associated retinopathy (CAR syndrome) with antibodies reacting with retinal, optic-nerve, and cancer cells. PMID- 2555715 TI - The Cushing syndromes: an enlarging clinical spectrum. PMID- 2555716 TI - The barrier to recombination between Escherichia coli and Salmonella typhimurium is disrupted in mismatch-repair mutants. AB - The requirement for DNA sequence homology in generalized genetic recombination is greatly relaxed in bacterial mutL, mutS and mutH mutants deficient in mismatch repair. In such mutants, intergeneric recombination occurs efficiently between Escherichia coli and Salmonella typhimurium, which are approximately 20% divergent in DNA sequence. This finding has implications for speciation, for regulating recombination between diverged repeated sequences, and for hitherto difficult interspecies hybridizations. PMID- 2555717 TI - Specific proteolysis of the c-mos proto-oncogene product by calpain on fertilization of Xenopus eggs. AB - The Xenopus c-mos proto-oncogene product, pp39mos, accumulates in the unfertilized egg during maturation, is hyperphosphorylated and exhibits protein kinase activity. On fertilization, or soon after the completion of meiosis, the accumulated pp39mos undergoes selective proteolysis. Using an in vitro protease assay system, we show here that this specific proteolysis is caused by the calcium-dependent cysteine protease, calpain. PMID- 2555718 TI - Release of endogenous GABA from the substantia nigra is not controlled by GABA autoreceptors. AB - The characteristics of the release of GABA from slices of the rat substantia nigra, elicited by electrical stimulation at frequencies of 0.5-48 Hz and by elevated K+ concentrations ranging from 15-35 mmol/l, was studied. Comparisons were made with cortical slices where the data were not available from previous studies. No GABA release could be evoked from rat nigral slices by electrical stimulation between 0.5 and 4 Hz, in contrast to cortical slices, in which this pool is sensitive towards inhibition by (-)-baclofen. Also, comparatively less GABA release could be evoked from nigral than from cortical slices by K+ concentrations between 15 and 25 mmol/l. While (-)-baclofen at 10 mumol/l inhibited release caused by 15 mumol/l K+ in cortical, it did not in nigral slices. GABA release caused by higher frequencies (8-48 Hz) or 30 mmol/l K+ concentrations was Ca2+-dependent and in the former case also tetrodotoxin sensitive. It had similar characteristics as in cortical slices and was insensitive towards (-)-baclofen, muscimol and bicuculline. Even more markedly than in the cortex, 30 mmol/l K+ released greater amounts of GABA than electrical stimulation at 24 Hz of a similar duration, suggesting the existence of one or several additional pool(s) of lesser excitability.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555719 TI - Modulation of noradrenaline and neuropeptide Y (NPY) release in the pig kidney in vivo: involvement of alpha 2, NPY and angiotensin II receptors. AB - The modulation of the release of noradrenaline (NA) and neuropeptide Y-like immunoreactivity (NPY-LI) was investigated in the pig kidney in vivo. Under control conditions a reproducible co-release of NA and NPY-LI was obtained upon stimulation of the renal nerves with 5 Hz for 1 min. Infusion of peptide YY (PYY, 1 microgram/kg/min i.v.), which binds to NPY receptors, caused renal vasoconstriction and reduced the stimulation-evoked overflow of NA and NPY-LI by 24 +/- 4 and 33 +/- 11%, respectively (P less than 0.01). The PYY effect was reversible and was absent 1 h after the infusion. The alpha 2-adrenoceptor antagonist yohimbine (0.2 mg/kg i.v.) enhanced the overflow of NA and NPY-LI 2- to 3-fold. The angiotensin converting enzyme inhibitor captopril (5 mg/kg i.v.) did not significantly affect the overflow of NA or NPY-LI evoked by the nerve stimulation. Angiotensin II (0.5 microgram/kg/min i.v.), on the other hand, induced a reversible enhancement of the overflow of both NA and NPY-LI by 71 and 77%, respectively (P less than 0.01). Infusion of endothelin (0.2 microgram/kg/min i.v.), which reduced renal blood flow by a magnitude similar to that evoked by angiotensin II, did not significantly alter the nerve stimulation evoked overflow of NA or NPY-LI. None of the administered drugs did significantly affect the percentage reduction in renal blood flow evoked by nerve stimulation. It is concluded that the release of NA and NPY-LI from sympathetic nerves in the pig kidney is inhibited in parallel via activation of NPY receptors by PYY and via alpha 2-adrenoceptors by endogenous NA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555720 TI - The gastrointestinal prokinetic benzamide derivatives are agonists at the non classical 5-HT receptor (5-HT4) positively coupled to adenylate cyclase in neurons. AB - We have previusly shown that a non-classical 5-hydroxytryptamine (5-HT4) receptor mediates the stimulation of adenylate cyclase activity in mouse embryo colliculi neurons in primary culture. The pharmacological characteristics of this receptor exclude the possibility that it belongs to the known 5-HT1, 5-HT2 or 5-HT3 receptor types. Here we report that this 5-HT receptor can be stimulated by 4 amino-5-chloro-2-methoxy substituted benzamide derivatives. All these compounds have been reported to be potent stimulants of gastrointestinal motility and some of them are 5-HT3 receptor antagonists. The rank order of potency of these substituted benzamide derivatives in stimulating cAMP formation was: cisapride greater than BRL 24924 greater than 5-HT greater than zacopride greater than BRL 20627 greater than metoclopramide. The non-additivity of benzamide and 5-HT activities suggests that 5-HT and the substituted benzamide derivatives act on the same receptor. Only ICS 205930, a recognized 5-HT3 receptor antagonist, competitively antagonized the stimulatory effect of cisapride, zacopride and BRL 24924. However, its pKi (6-6.3) for this new receptor was very different from its pKi for 5-HT3 receptors (pKi = 8-10). Other selective 5-HT3 receptor antagonists with an indole group (BRL 43694 and GR 38032F), with a benzoate group (cocaine, MDL 72222) or with a piperazine group (quipazine) were ineffective in reversing the stimulatory effect of benzamide derivatives. Exposure of neuronal cells to potent agonists at this receptor such as BRL 24924 rapidly reduces its capacity to stimulate cAMP production.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555721 TI - The motor response to ethylenediamine of the rat isolated duodenum: involvement of GABAergic transmission? AB - We have studied the motor response to ethylenediamine (EDA), a well known releaser of endogenous GABA, on the longitudinal muscle of the rat isolated proximal duodenum in presence of atropine (3 microM) and guanethidine (3 microM). EDA produced a concentration-(0.03-3 microM)-dependent relaxation which was potentiated when the preparations were exposed to GABA during the equilibration period. The GABA-induced potentiation of the response to EDA was prevented by nipecotic acid, an inhibitor of GABA uptake. The response to EDA was partially inhibited by 3-mercaptopropionic acid, a known inhibitor of GABA release. However, contrary to the relaxant response produced by exogenous GABA, the EDA induced relaxation was picrotoxin-(0.1 microM)-resistant. In preparations pre exposed to GABA, the response to EDA was partially tetrodotoxin-(1 microM) sensitive. By contrast, in preparations not exposed to GABA, the EDA-induced relaxation was totally tetrodotoxin resistant. The response to EDA was abolished or largely inhibited in preparations excised from rats in which the proximal duodenum was chemically denervated by exposure (2 weeks before), to benzalkonium chloride (BZK). Likewise, the indirect relaxations produced by electrical field simulation. DMPP, capsaicin or GABA were abolished by BZK pretreatment while noradrenaline was still effective. These findings indicate that the relaxant response to EDA is neurogenic in origin, while being largely tetrodotoxin resistant. A GABAergic mechanism could be involved but also other inhibitory transmitter(s) should be taken into account. EDA appears a useful tool to study the inhibitory non-adrenergic non-cholinergic innervation of the rat proximal duodenum. PMID- 2555722 TI - Two phases of the prostaglandin F2 alpha-induced contraction in guinea-pig taenia coli involve different Ca2+ channels. AB - Properties of the contraction produced by PGF2 alpha in the guinea-pig taenia coli were compared to those produced by ACh. Prostaglandin (PG) F2 alpha (3 x 10( 7) M) and acetylcholine (ACh, 10(-5)M) induced an initial transient contraction (phasic contraction) and a subsequent late contraction (tonic contraction). Both phasic and tonic contractions produced by PGF2 alpha or ACh were abolished in Ca2+ -free Krebs solution containing 0.5 mM EGTA. The tonic contractions caused by PGF2 alpha and ACh were markedly suppressed by alpha-[3-[[2-(3,4-dimethoxy phenyl)-ethyl]-methylamino]-propyl]- 3,4,5-trimethoxy-alpha-(1 methylethyl)benzeneacetonitrile hydrochloride (D600, greater than 10(-7)M) as well as nifedipine (5 x 10(-9)M), a Ca2+-antagonist. However, the phasic contraction produced by PGF2 alpha, but not by ACh, was greatly inhibited by Mn2+ (greater than 10(-4)M). Furthermore, the phasic contraction caused by PGF2 alpha was abolished in 18 mM K+ Krebs solution with D600 (2 x 10(-7)M), whereas that induced by ACh and the tonic contractions produced by PGF2 alpha as well as by ACh were unaffected in this high K+ solution without D600. Membrane potentials of the tissue in normal (K+, 5.9 mM) and 18 mM K+ Krebs solution containing D600 were about -55 mV and -43 mV, respectively. In a fluorescence study which used Fura-2 an intracellular free Ca2+ indicator in the presence of D600, PGF2 alpha and ACh increased fluorescence intensity in the tissue, which coupled with the magnitude of contractions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555724 TI - Cyclic GMP stimulation by vasopressin in LLC-PK1 kidney epithelial cells is L arginine-dependent. AB - The L-arginine antagonist NG-monomethyl-L-arginine has been shown to inhibit nitric oxide formation from L-arginine in endothelial cells. In the present study NG-monomethyl-L-arginine was used to assess the role of L-arginine for cyclic GMP stimulation by vasopressin in a kidney epithelial cell line (LLC-PK1). Preincubation of cells with 1 mumol/l, 10 mumol/l and 100 mumol/l NG-monomethyl-L arginine decreased cyclic GMP stimulation at 1 mumol/l vasopressin by 25%, 71% and 90%, respectively. This inhibition by NG-monomethyl-L-arginine was markedly reduced by L-arginine (2 mmol/l) but not D-arginine (2 mmol/l). Cyclic GMP stimulation by the calcium ionophore A23187 was also inhibited by NG-monomethyl-L arginine and enantioselectively restored by L-arginine. However, NG-monomethyl-L arginine did not affect cyclic GMP stimulation by sodium nitroprusside that spontaneously releases nitric oxide. These results suggest that, in kidney epithelial cells, vasopressin induces nitric oxide formation from L-arginine leading to activation of soluble guanylate cyclase. It is concluded that nitric oxide formation from L-arginine is not only responsible for endothelium-dependent relaxation but may be a more general pathway with regulatory function for intracellular guanylate cyclase activity. PMID- 2555723 TI - Pharmacological properties of voltage-dependent calcium channels in functional microvessels isolated from rat brain. AB - Voltage-operated calcium channels were studied in rat intracerebral microvessels. The contractile reactivity to KCl-depolarization was assessed by the measurement of internal diameter of superfused microvessels. Dihydropyridine receptor sites associated with calcium channels were identified and characterized using 3H(+)PN 200-110 [isopropyl-4-(2,1,3-benzodiazol-4-yl)-1,4-dihydro-2,6-dimethyl-5- methoxycarbonyl-pyridine-3-carboxylate]. Depolarization induced by high-KCl solution produced a marked reduction of the internal diameter of cerebral microvessels which was associated with the appearance of rhythmic activity. The vessel contraction was reversible and abolished by nimodipine. Binding studies with 3H(+)PN 200-110 revealed the existence of a single class of specific, stereoselective and voltage-dependent binding sites which bound (+)PN 200-110 with a KD of 88 +/- 6.6 pmol l-1 at 37 degrees C in microvessels incubated in NaCl medium. When microvessels were incubated in KCl-medium, the apparent KD value was reduced to 35 +/- 2 pmol l-1. Bmax was not significantly changed. The effect of KCl was not related to concomitant changes in the Na concentration. The potency of various dihydropyridine derivatives in inhibiting 3H(+)PN 200-110 binding was in agreement with their pharmacological potency in smooth muscle preparations. The effect of PN 200-110 and of nimodipine was stereoselective. Ki values of PN 200-110 and of nimodipine were increased in depolarized preparations, while nifedipine's potency was unchanged. Verapamil was only a partial inhibitor of 3H(+)PN 200-110 binding. The effect of diltiazem was stereoselective: the (+)-cis isomer enhanced the binding and the (-)-cis isomer of diltiazem poorly inhibited the binding of PN 200-110. Results showed that isolated cerebral microvessels possess functional voltage-operated calcium channels, which contain potential-modulated receptors for dihydropyridine calcium entry blockers with characteristics similar to those described in other tissues. PMID- 2555725 TI - [Irritable bowel syndrome]. PMID- 2555726 TI - [Partial liver resection]. PMID- 2555727 TI - [Acute Prussic acid poisoning in an infant]. PMID- 2555728 TI - [The role of dietary fiber in gastroenterology: fact or fancy?]. PMID- 2555729 TI - Treatment of small cell carcinoma of the lung. PMID- 2555730 TI - Unusual development of polyoma virus in the brains of two patients with the acquired immune deficiency syndrome (AIDS). AB - Two HIV-positive male patients presented with a variety of symptoms including hemiparesis, unsteadiness, progressive loss of vision and poor memory. There were multiple non-enhancing lesions shown by CT scan in the white matter of the cerebral hemispheres. Specimens obtained by burr-hole biopsy showed the features of progressive multifocal leucoencephalopathy (PML) in both cases. Electron microscopy demonstrated round and rod shaped particles of papovavirus in the nuclei and cytoplasm of oligodendrocytes and in processes of astrocytes where abnormal and florid modes of viral replication were seen. Additional features observed were viral particles suggestive of an enterovirus in Case 1 and, in both specimens, massive membrane proliferation within both nuclei and cytoplasm of infected cells together with the presence of tubuloreticular structures (TRS) in the cytoplasm of endothelial cells. At post-mortem, the brain of patient 2 showed PML and HIV encephalitis. The presence of HIV was confirmed by immunohistochemical methods. We suggest that in AIDS patients the abnormality of the immune system induced by HIV causes abnormal replication patterns of papovavirus in the brain. In addition, these cases confirm the frequent occurrence in AIDS patients of TRS, now considered to be a marker for HIV. PMID- 2555731 TI - Metastatic small cell carcinoma to the pineal body: case report. AB - A case of metastatic small cell carcinoma to the pineal body is reported and the clinical and radiological features of this rare tumor are discussed. A 66-year old man presented with progressive dementia, gait disturbance, vertical gaze palsy, and convergence retraction nystagmus. Computed tomographic scan revealed a 2 X 3-cm high-density mass in the pineal region, which showed strong contrast enhancement. The tumor was resected through a right occipital transtentorial approach. The pathological diagnosis of the surgical specimens was small cell carcinoma. A systemic workup for the primary lesion subsequently revealed small cell carcinoma of the lung. Although occurring rarely, metastatic tumor should be considered in the differential diagnosis of pineal tumor in elderly patients. PMID- 2555732 TI - Brain metastasis from nonseminomatous germ cell tumors of the testis: case report and review of the role of surgery. AB - The prognosis for patients with nonseminomatous germ cell tumor of the testis is good, even when extensive metastatic disease is present, because this tumor is very sensitive to chemotherapy with cisplatin, vinblastine, and bleomycin (PVB). If a metastasis occurs in the brain, however, the prognosis is poor because the blood-brain barrier limits the entrance of these drugs into the brain and creates a sanctuary for tumor. The current treatment for a brain metastasis is either standard PVB chemotherapy plus whole brain radiation therapy or a rigorous chemotheraputic regimen that penetrates the blood-brain barrier better than PVB. Surgery is seldom used for brain metastasis, largely because of the poor results with surgical debulking in noncentral nervous system disease. This is the report of a patient with disseminated nonseminomatous germ cell tumor and multiple large brain metastases, who was treated with surgery, PVB, and whole brain radiation therapy and cured. Evidence is presented to support a role for surgical debulking in patients with large brain metastasis. PMID- 2555733 TI - Release and modulation of release of serotonin in rabbit superior colliculus. AB - The release of previously incorporated [3H]serotonin and its presynaptic modulation were studied in slices of rabbit superior colliculus. Electrical stimulation at frequencies of 0.017-3 Hz greatly increased the outflow of tritiated compounds; this response was almost abolished by tetrodotoxin and in a low calcium medium. Unlabelled serotonin, when added in the presence of nitroquipazine, an inhibitor of high-affinity neuronal serotonin uptake, reduced the electrically evoked overflow of tritium, an effect antagonized by metitepin. Given alone, metitepin caused an increase. The evoked overflow was also decreased by clonidine, and the effect of clonidine was counteracted by phentolamine. Phentolamine itself increased the overflow response. However, this was probably not due to antagonism against an inhibitory effect of endogenous noradrenaline because, first, the selective alpha 2-adrenoceptor antagonist idazoxan did not share with phentolamine the overflow-enhancing effect, second, phentolamine continued to increase the overflow after noradrenergic axons had been destroyed by 6-hydroxydopamine, and third, the facilitatory effects of metitepin and phentolamine were not additive. Phentolamine, like metitepin, antagonized the presynaptic inhibitory effect of serotonin, indicating that it may increase the evoked overflow of tritium by blocking serotonin receptors rather than alpha adrenoceptors. Ethylketocyclazocine decrease the electrically evoked overflow, and its effect was prevented by naloxone: peptides selective for opioid mu- or delta-receptors caused no change. Nicotine increased the basal outflow of tritium (in the absence of electrical stimulation); the increase was attenuated by hexamethonium and low calcium medium. No or minimal changes in tritium outflow were obtained with beta-adrenoceptor, dopamine receptor, muscarine receptor and GABA receptor ligands or with substance P and glutamate. In conjunction with our previous studies, these results indicate that serotonin is a neurotransmitter in the superior colliculus. Its release is modulated through presynaptic autoreceptors (probably 5-HT1), alpha 2-adrenoceptors, opioid kappa-receptors and nicotine receptors, of which only the autoreceptors receive an endogenous input, at least under the experimental conditions chosen. Each of the three groups of collicular monoamine axons that we have studied recently (cholinergic, noradrenergic, serotoninergic) possesses a specific pattern of presynaptic, release-modulating receptors. A physiological role seems likely only for the alpha 2-autoreceptors at the noradrenergic and the 5-HT1-autoreceptors at the serotoninergic axons. PMID- 2555734 TI - Immunocytochemical demonstration of opioid receptors in selected rat brain areas and neuroblastoma x glioma hybrid (NG108-15) cells using a monoclonal anti idiotypic antibody. AB - A monoclonal anti-idiotypic opioid receptor antibody was used for the light microscopic visualization of opioid receptors in several brain structures and monolayer cultures of a neuroblastoma x glioma hybrid cell-line (NG108-15). The antibody proved to be specific, displaying affinity for mu greater than delta much greater than kappa opioid receptors. Receptor distribution in the brain areas studied was in agreement with previous autoradiographic analyses; of particular interest, high densities of immunoreactive opioid receptors were found in the perikarya and in the initial parts of the axons and dendrites; light microscopy did not allow an exact determination of the subcellular localization of opioid receptors, but the immunoreactivity seemed to be associated with the plasma membrane and to be present within the cytoplasm as well. Similar observations were made for the cell bodies and neurites of NG108-15 cells. The methodology described potentially permits the study of opioid receptor distribution in discrete brain areas under different physiological and pharmacological conditions and of the ontogeny of these receptors; in addition, it may help to find a morphological basis for events such as receptor internalization and recycling. PMID- 2555736 TI - Quantal transmission at Mauthner axon target synapses in the goldfish brainstem. AB - The Mauthner axon contacts two principal neurons within the brainstem: the cranial relay neuron and the follower neuron. The cranial relay neuron is excited through axoaxonic contacts by both Mauthner axons, while the follower neuron is excited by the Mauthner axon ipsilateral to its soma and inhibited (through an interposed neuron) by the contralateral Mauthner axon. We have investigated the properties of these two Mauthner axon target synapses with simultaneous pre and postsynaptic recordings in order to determine if transmission can be described in terms of the quantal hypothesis and to determine what quantal parameters change with alterations in the frequency of Mauthner cell activation. Small depolarizing postsynaptic potentials, recorded in the cranial relay neurons, increase their frequency of occurrence when depolarizing currents are applied to the Mauthner axon, without changing their mean amplitude, indicating that they arise from the Mauthner axon terminal and represent quantal units of the evoked cranial relay neuron excitatory postsynaptic potentials. Similar small, spontaneous potentials can also be recorded from the follower neurons. Amplitude histograms of evoked postsynaptic potentials (in both cranial relay neurons and follower neurons) can be fit by binomial models based on the amplitude and variance of the spontaneous potentials, suggesting that the quantal hypothesis is a valid descriptor of synaptic transmission at these contacts. Quantal content ("m"), the number of releasable quanta ("n"), and the probability of release ("p") are relatively high for both the cranial relay neurons and follower neurons. Increasing the frequency of Mauthner axon stimulation results in a decrement in the postsynaptic potential amplitude in both target cells. There is no change in the amplitude of the small, spontaneous potentials (measured during the stimulation period) with stimulus frequency, indicating a decrease in quantal content, but not quantal size. The change in quantal content is explained in binomial terms by a decrease in the number of releasable quanta, but not the probability of release in both the cranial relay neurons and the follower neurons. These findings suggest that the supply of transmitter available to be released decreases with increasing stimulus frequency without affecting the nerve terminal's ability to release transmitter (at these stimulus frequencies). Synaptic vesicles, observed in clusters in the terminals at the ultrastructural level, may be a physical correlate of the binomial parameter ("n"). It is predicted that the decrease in the binomial parameter ("n") with increasing frequency of stimulation is accompanied by a decrease in the number of active zones occupied by vesicles.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2555735 TI - Properties of excitatory amino acid receptors on sustained ganglion cells in the cat retina. AB - Iontophoretic effects of N-methyl-D-aspartate, quisqualate and kainate and a variety of excitatory amino acid receptor antagonists, on retinal ganglion cells, were studied in optically intact eyes of barbiturate anesthetized cats. All three agonists raised the spontaneous firing of both ON- and OFF-sustained retinal ganglion cells, with the potency order of kainate much greater than quisqualate greater than N-methyl-D-aspartate. However, the excitatory amino acid analogues readily saturated the receptors and reduced the visually driven firing of cells with high spontaneous firing, but mimicked an increase in endogenous excitatory amino acid release and raised the visually induced response in cells with low spontaneous firing. The quinoxaline compound, 6-cyano-2,3 dihydroxy-7 nitroquinoxaline and 6-7-dinitroquinoxaline-2,3-dione, blocked the visually driven firing and kainate- and quisqualate-induced excitation, whilst 3[+)-2 carboxypiperazin-4-yl)propyl-1-phosphonate, antagonized the N-methyl-D-aspartate induced excitation, but failed to block visually driven firing of the retinal ganglion cells. The broadband excitatory amino acid receptor antagonists, such as kynurenate, were also effective in antagonizing the visually driven response and also blocked the N-methyl-D-aspartate- as well as kainate- and quisqualate induced responses. These results suggest that the receptors at the bipolar/ganglion cell synapse are of the non-N-methyl-D-aspartate type, but that N-methyl-D-aspartate receptors are also present on ganglion cells although their physiological role is unclear. PMID- 2555737 TI - Dissociation between changes in glutamate receptor binding sites and their coupling to phosphatidylinositol metabolism following intrahippocampal colchicine injection. AB - Intrahippocampal colchicine injection produces a rapid death of granule cells and pyramidal neurons in the hippocampus in the rat. Under the appropriate assay conditions, [3H]glutamate labels the N-methyl-D-aspartate type of glutamate receptors while [3H]alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate labels the quisqualate type. Unilateral injection of colchicine (15 micrograms) in the dorsal hippocampus did not produce any change in [3H]glutamate and [3H]alpha amino-3-hydroxy-5-methylisoxazole-4-propionate binding in membrane fractions from the dentate gyrus or CA1 field contralateral to the injection side, at least up to 12 days after the injection. However, it produced a progressive decrease in the binding of both ligands in dentate gyrus and CA1 of the injected hippocampus. In the dentate gyrus the changes in binding as a function of time after the injection were biphasic with a rapid exponential decrease (t1/2 about 8 days for both [3H]glutamate and [3H]alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate) until 12 days after the injection followed by a much slower decrease afterwards. A similar pattern was observed in CA1 although the changes in binding were smaller and delayed by about three days as compared to the dentate gyrus. Kinetic analyses of the binding at equilibrium were performed seven days after the injection and indicated that the changes in [3H]glutamate binding were due to a change in the maximum number of sites but not in affinity for the ligand.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555738 TI - GABAA receptor-mediated chloride flux in brain homogenates from rat lines with differing innate alcohol sensitivities. AB - Muscimol stimulation of 36Cl- flux through GABAA receptor-associated ion channels was compared in combined cerebral cortical and cerebellar homogenates from two lines of rats produced by selective outbreeding for high and low alcohol sensitivities. There was no difference in the muscimol effects between ethanol naive alcohol-sensitive and alcohol-insensitive rats. Acute administration of ethanol (2 g/kg, i.p.) and lorazepam (3 mg/kg, i.p.) significantly reduced the percentage stimulation by muscimol in the alcohol-sensitive animals. The results suggest that genetic selection towards differences in the sensitivity to motor impairing effects of moderate ethanol doses does not produce alterations in the direct agonist-induced GABAA receptor function. This receptor function was, however, down-regulated in the alcohol-sensitive rats by acute ethanol and benzodiazepine treatments, indicating the involvement of GABAergic activity in the mechanisms of, or in the neural adaptations to, acute intoxication in genetically sensitive animals. PMID- 2555739 TI - Distribution of alpha latrotoxin receptor in the rat brain by quantitative autoradiography: comparison with the nerve terminal protein, synapsin I. AB - Tissue slice autoradiography was employed to reveal the brain distribution of the receptor for alpha latrotoxin, the presynaptic neurotoxin of the black widow spider venom. The receptor distribution pattern was compared with that of a marker protein for nerve endings, synapsin I, a phosphoprotein known to be present within nerve terminals. The alpha latrotoxin receptor and synapsin I were detected in gray matter-containing regions but their relative amounts were not constant. In the cerebral cortex and in the caudatum their distribution was similar, while in the hippocampus they were both abundant, but their distribution varied: synapsin I labeling was heavier in CA4 and CA3, alpha latrotoxin receptor labeling in CA1 and dentate gyrus. A dissociation was also observed in the globus pallidus and in the lateral thalamic nuclear complex, where alpha latrotoxin receptor labeling was very weak. The most striking dissociation occurred in the cerebellum, where the molecular layer was strongly labeled for synapsin I, but almost unlabeled for the alpha latrotoxin receptor, which was more concentrated in the granular layer. Taken as a whole, the data appear compatible with a widespread localization of the alpha latrotoxin receptor at synapses. However, they also suggest that either some nerve terminals are insensitive to alpha latrotoxin, or the receptor for the toxin is not present at a similar concentration in all presynaptic plasma membranes. PMID- 2555740 TI - Distribution of the omega-conotoxin receptor in rat brain. An autoradiographic mapping. AB - The distribution of [125I]omega-conotoxin GVIA binding sites, the putative voltage-sensitive calcium channels, was studied by an autoradiographic method in the rat brain. The toxin binding sites were distributed throughout the brain in a highly heterogeneous manner. The highest density of the binding sites was observed in the cerebral cortex, hippocampus, amygdaloid complex, substantia nigra, caudate putamen, superior colliculus, nucleus of the solitary tract, and the dorsal horn of the cervical spine. The glomerular layer of the olfactory bulb, molecular layer of the cerebellar cortex, and posterior lobe of the hypophysis showed intermediate density but the density was higher than in the surrounding areas. The globus pallidus, thalamic areas, inferior olive, and pontine nuclei showed low density, while no binding sites were observed in the white matter tract regions such as the internal and external capsule, corpus callosum, fimbria of the hippocampus, fornix, stria medullaris of the thalamus, and fasciculus retroflexus. This distribution of omega-conotoxin binding sites indicates that the toxin binding sites are localized in those areas of the brain enriched in synaptic connections. This distribution pattern resembles that reported for voltage-sensitive sodium channels but it differs from that of the binding sites of dihydropyridines and verapamil. These results suggest that omega conotoxin recognizes different molecules from organic calcium channel antagonist binding sites and that omega-conotoxin-sensitive voltage-sensitive calcium channels are concentrated in the synaptic zones and play a key role in the excitation-secretion coupling of neurotransmitters. PMID- 2555741 TI - Nodose ganglionectomy selectively reduces muscarinic cholinergic and delta opioid binding sites in the dorsal vagal complex of the cat. AB - The dorsal vagal complex of the medulla oblongata, comprising the nucleus tractus solitarii, the area postrema and the dorsal motor nucleus of the vagus nerve, is an important brainstem regulatory center for the autonomic nervous system. The major afferent input from abdominal and thoracic viscera to this region is via vagal sensory neurons which have their cell bodies in the nodose ganglion. Autoradiography has been used to study the effects of unilateral nodose ganglionectomy on receptor binding sites in this region of the brain for the neurotransmitters acetylcholine, norepinephrine, and opioids. Nodose ganglionectomy had no discernible effect on alpha 2 noradrenergic ([3H]p aminoclonidine) or mu opioid [( 3H]Tyr-D-Ala-Gly-(NMePhe)-Gly-ol) binding sites. However, ganglionectomy did produce a 25% decrease in [3H]quinuclidinyl benzilate (muscarinic cholinergic) binding in the subnucleus gelatinosus of the solitary nucleus, and a marked decrease in [3H][D-Pen5]enkephalin (delta opioid) binding in the dorsomedial subnucleus of the nucleus tractus solitarii, ipsilateral to the lesion. These data suggest that muscarinic cholinergic and delta opioid receptors may be present on terminals of vagal afferent neurons that project to these specific brainstem regions. Since these vagal afferent neurons are known to arise, at least in part, from the gastrointestinal tract, it is possible that cholinergic and/or opioid receptors modulate specific autonomic functions associated with gastric sensory information such as satiety or nausea and emesis. PMID- 2555742 TI - Differences in the chemical expression of rat primary afferent neurons which innervate skin, muscle or joint. AB - The fluorescent dye Fast Blue was injected in anaesthetized rats into either skin, muscle or knee joint of the hindlimb. Following retrograde transport of the dye to lumbar dorsal root ganglia, the cell bodies of primary afferent neurons innervating these different target tissues were identified in ganglion sections by fluorescence microscopy. The sections were processed to demonstrate activity of the enzyme thiamine monophosphatase, or immunoreactivity to calcitonin gene related peptide, substance P, or somatostatin, in Fast Blue labelled neurons. In all cases immunoreactivity to the antineurofilament antibody RT97 was used to classify dorsal root ganglion cells as being either small dark (RT97 negative, unmyelinated axons) or large light (RT97 positive, myelinated axons). The proportion of small dark cells labelled from each target decreased in the order: skin, muscle, joint. Thiamine monophosphatase and somatostatin were present only in small dark cells, while calcitonin gene-related peptide and substance P were found in both small dark and large light cells. In large light cells of all three targets, more contained calcitonin gene-related peptide than substance P. Among small dark cells, thiamine monophosphatase and somatostatin were found predominantly in skin afferents, while calcitonin gene-related peptide and substance P were more common in muscle and joint afferents. The chemical expression of primary afferents is therefore characteristic of the peripheral target they innervate. This could reflect either a maintained influence of the target on the afferents, or the factors which operate only during development. PMID- 2555744 TI - Case for diagnosis. Angiomatoid malignant fibrous histiocytoma. PMID- 2555743 TI - Chronic opiate receptor activation in vivo alters the level of G-protein subunits in guinea-pig myenteric plexus. AB - Studies with the longitudinal muscle-myenteric plexus preparation of the guinea pig ileum were undertaken to investigate the relationship between guanine nucleotide-binding proteins (G-proteins) and chronic opioid receptor activation in vivo. Treatment with the narcotic agonist fentanyl, at doses which render the preparation tolerant and dependent, led to an increase of pertussis toxin catalysed incorporation of ADP-ribose in a protein of approximately 40,000 mol. wt. Quantitative immunoblotting, using site-directed antisera, demonstrated an upregulation of G alpha i/G alpha o and, to an even greater degree, of G beta. However, the level of G alpha s was decreased by the same treatment. All alterations observed were abolished by the concomitant presence of the antagonist naloxone. The implications of this differential regulation of G-protein subunits for opiate-induced tolerance and dependence are discussed. PMID- 2555745 TI - Granular cell tumor of the breast. AB - Three cases of granular cell tumor of the breast are reported. Granular cell tumor of the breast is a rare benign tumor with a malignant appearance both clinically, radiologically and macroscopically. The cytology is quite characteristic although the tumor clinically often resembles a carcinoma. Frozen section can be misinterpreted, unnecessary mastectomy or axillary dissection can be prevented by careful cytological examination. PMID- 2555746 TI - Functional expression of a subtype of the Ca2+ channels in the embryonic rat hippocampus as detected by dual-fluorescence flow cytometric analysis. AB - We have analyzed the expression of the Ca2+ channels in hippocampal cell suspensions from the 18- to 20-day-old rat embryo using dual-fluorescence flow cytometry. A high concentration of K+ induced elevation of the cytoplasmic free Ca2+ concentration ([Ca2+]i) as well as membrane depolarization. The high K+ evoked [Ca2+]i increase was inhibited by phenytoin, but not by either nifedipine or nicardipine. These agents had no effect on the high K+-induced membrane depolarization. These findings suggest that a subtype corresponding to the low voltage-activated Ca2+ channel is expressed in the embryonic rat hippocampal cells. PMID- 2555747 TI - Loss of N-methyl-D-aspartate sensitivity of cerebellar Purkinje cells after climbing fiber deafferentation. An in vivo study in the rat. AB - The sensitivity of cerebellar Purkinje cells to brief iontophoretic applications of excitatory amino acids has been studied in vivo in rats treated from 15 days to 4 months beforehand with 3-acetylpyridine in order to destroy the inferior olive. Responses of Purkinje cells (PCs) chronically deprived of climbing fibers were thus investigated using extracellular microelectrodes and compared to those of a group of control rats. No changes in the relative efficiencies of L glutamate, L-aspartate, quisqualate and kainate have been observed. In contrast, the excitations induced by N-methyl-D-aspartate (NMDA) on most PCs in control animals, were no longer present after climbing fiber deprivation. These results show that both NMDA and non-NMDA receptors are present on PCs of adult rodents and that the NMDA responses are strongly depressed when PCs are deafferented of the climbing fibers. PMID- 2555748 TI - 2',3'-cyclic nucleotide-3'-phosphodiesterase activity as an index of myelin in the post-mortem brains of patients with Alzheimer's disease. AB - The activity of 2',3'-cyclic nucleotide-3'-phosphodiesterase (CNPase) was determined as an index of myelin in the post-mortem brain samples of 16 patients with Alzheimer's disease (AD) and of 14 controls. The CNPase activity was mildly increased in the temporal, hippocampal and parietal cortex in AD subjects pointing to a relative sparing of myelin as compared to the neuron loss within cortex in AD. In the hippocampus the CNPase activity was decreased in AD patients indicating loss of myelin and thus myelinated axons in this area in AD. PMID- 2555749 TI - Increased alpha 2- and beta 2-adrenergic receptors in cerebral microvessels in Alzheimer disease. AB - Adrenergic receptors exist in brain microvessels which are innervated by noradrenergic locus ceruleus neurons. Biochemical and pathological studies indicate locus ceruleus degeneration in Alzheimer disease (AD), which can cause adrenergic receptor alterations in brain microvessels. To assess this, we studied adrenergic receptors in human brain microvessels from AD subjects and age-matched controls by ligand binding methods. Total beta receptors of cerebral microvessels and beta 2 receptors, the type which predominates in microvessels, were significantly increased in AD. Compared to the cerebral cortex, there was a paucity of alpha 1-adrenergic receptors in cerebral microvessels, and they did not change in AD. Binding to alpha 2 receptors in cerebral microvessels was approximately 50% of that in the cortex, and these receptors increased by approximately 60% in cerebral microvessels of AD subjects. These findings suggest adrenergic receptor 'upregulation' in response to noradrenergic deafferentation in AD, which may have functional consequences at the blood-brain barrier. PMID- 2555750 TI - Differential distribution of GABAA receptor mRNAs in bovine cerebellum- localization of alpha 2 mRNA in Bergmann glia layer. AB - Using in situ hybridization histochemistry, we have demonstrated that 3 alpha subunit mRNAs of the GABAA receptor are present in different cell populations of the bovine cerebellum. While the alpha 1 mRNA is the most abundant and is present in granule cells, Purkinje cells and stellate/basket cells, the alpha 2 mRNA appears to be confined to the Bergmann glial cell layer. The alpha 3 mRNA is only expressed in the Golgi cells. This differential distribution of GABAA receptor mRNA subtypes suggests a previously unrecognized complexity of GABAergic transmission in the cerebellum. PMID- 2555751 TI - PNMT-containing neurons in the rostral medulla oblongata (C1, C3 groups) are transneuronally labelled after injection of herpes simplex virus type 1 into the adrenal gland. AB - Herpes simplex virus type 1 (HSV1) was injected into the rat adrenal gland. After 3 days the rat CNS was processed immunohistochemically to demonstrate viral antigen. In the lower thoracic spinal cord viral antigen was found in neurons in the intermediolateral column. In the medulla oblongata HSV1-positive neurons were found in the raphe pallidus and in the C1 and C3 regions of the rostral medulla. Approximately 50% of HSV1-positive neurons in the C1 and all the HSV1-positive neurons in the C3 area also contained phenylethanolamine N-methyltransferase (PNMT) and were thus identified as C1 and C3 cells. The HSV1-positive neurons in the C1 region which did not contain PNMT were also negative for tyrosine hydroxylase and were therefore not catecholamine-synthesizing neurons. The HSV1 positive neurons in the medulla oblongata were presumably transsynaptically labelled from the adrenal gland and our study therefore provides neuroanatomical evidence supporting the view that some C1 neurons are involved in controlling the function of the adrenal gland. PMID- 2555752 TI - The management of diaphragmatic lesions in ovarian carcinoma. AB - Cytoreduction is currently an essential feature of the treatment of carcinoma of the ovary. Occasionally, optimal reduction is achieved except for localized diaphragmatic lesions. Two patients who were treated with primary cytoreduction for epithelial ovarian carcinoma had localized full-thickness diaphragmatic metastasis. Often resection is avoided because of concern about pneumothorax. A safe and easy method is described for removing peritoneal and diaphragmatic muscular metastasis without the need for a thoracostomy tube. These patients were cytoreduced without morbidity. The first patient is currently free of disease at 17 months after surgery. The second patient had a 13-month disease-free interval before her abdominal recurrence. PMID- 2555753 TI - Human papillomavirus DNA in fomites on objects used for the management of patients with genital human papillomavirus infections. AB - We investigated the possibility that human papillomavirus (HPV) DNA could be present on objects that are used for the management of patients with genital HPV infections. Human papillomavirus DNA was identified by filter hybridization in swabs taken from eight of 16 (50%) of surgical gloves. Similarly, HPV DNA was found in 23 of 62 (37%) and one of 62 (1.6%) of biopsy forceps before and after sterilization in 30% tincture of Savlon for 30 minutes, respectively. Five of 22 (23%) and one of 22 (4.5%) of cryoprobe tips tested positive for HPV DNA before and after cleaning with 90% ethanol solution for 1 minute, respectively. The rate of HPV DNA positivity after sterilization is low. Whether HPV on fomites is infectious was not evaluated in this study. Washing and soaking them in detergents containing 2% glutaraldehyde or heating them to 100C or higher should be adequate. Use of separate gloves for manual examination of the external and internal anogenital tract is advisable. PMID- 2555754 TI - [Characteristics of visual fatigue developing during the performance of precision operations]. AB - The assessment of visual fatigue developing in the process of performing precision works with the usage of monocular optic devices has shown that visual fatigue develops due to impairment of the functional state of binocular functions to which changes of the accommodation system join. The obtained psychophysiologic portraits can be helpful for a quantitative assessment of developing visual fatigue. PMID- 2555755 TI - [The efficacy of antiviral preparations in treating epidemic keratoconjunctivitis]. AB - A study of the therapeutic effect in 159 patients with epidemic keratoconjunctivitis treated by antiviral preparations: alpha-interferon, prodigiosane, 0.25% phlorenale and 0.05% riodoxole ointments has a sufficiently high effect of the mentioned preparations; the duration of the disease is shorter in these patients as compared with patients treated by common antibiotic preparations. As compared with other medicinal preparations, the therapeutic effect of alpha-interferon and phlorenale was evident not only in the shortening of terms of treatment, but also in prevention of keratitis in such patients. The therapeutic effect was higher when the preparations were prescribed at early terms (1-4 days) of the disease. PMID- 2555756 TI - [Collalysine treatment of intraocular hemorrhages]. AB - For treatment of 35 patients with intraocular hemorrhages (22 persons with hemophthalmos and 13 persons with hemorrhages into the retina) collalysin was injected subconjunctivally, 50 CU in a 0.3 ml of 0.5% novocaine solution. A course of treatment consisted of 10-12 injections. A control group included patients treated by fibrolysin injections (22 persons). Collalysin proved to be more effective: initial visual acuity from light perception to 0.03 increased to 0.1 and higher in 50% of patients and to 0.4-0.7 in 22.7%. Thus, subconjunctival injections of collalysin in treatment of intraocular hemorrhages stimulate its effectiveness. PMID- 2555757 TI - [Prognostic factors in small cell bronchial cancer]. AB - 199 cases with small cell lung cancer, which appeared in our hospital records between 1980 and 1981, and which could be followed up to the end of 1987, were retrospectively analyzed. For our analysis of the evaluation of prognostic factors we utilized the Kaplan-Meier system as well as the Log-Rank test. Factors of prognostic significance were: sex, stage of disease, the N-status as well as the M-status. PMID- 2555758 TI - [In vivo imaging of Hodgkin's lymphomas using monoclonal antibodies]. AB - The Hodgkin- and Reed-Sternberg cell associated monoclonal antibody HRS-1 was labeled with radioactive iodine and injected into 6 patients with Hodgkin's lymphoma for immunoscintigraphic imaging. Four of five patients who received 131I labeled HRS-1 had a positive immunoscintigram. In the sixth patient, the HRS-1 Mab was labeled with 123I in order to utilize tomoscintigraphy instead of linear scintigraphy. Bony disease was demonstrated by immunoscintigraphy in this patient. These preliminary results demonstrate that immunoscintigraphy with iodine-labeled HRS-1 Mab is feasible and informative in Hodgkin's lymphoma. The real clinical value of immunoscintigraphy in Hodgkin's lymphoma must be determined in a larger series of patients. Several modifications of the technique such as the use of Fab or F(ab')2 fragments should further improve the results. PMID- 2555759 TI - Modified humoral leukocyte adherence inhibition test for detection of lung cancer. AB - The freshly separated indicator cells (suspension of leukocytes) used in humoral leukocyte adherence inhibition test were labeled either with 14C-amino acid mixture or 3H-concanavalin A (3H-ConA). Instead of counting the adherent cells, the amount of 'adherent' radioactivity was measured by a liquid scintillation spectrometer. By the modified method, sera from 25 lung-carcinoma-bearing patients as well as serum samples from 21 healthy persons were examined in the presence of crude antigens prepared from 'normal' lung tissue or lung tumors of various histologic types. Although the results demonstrated high specificity and reproducibility of both methods, the binding of 3H-ConA to the surface of adherent cells is more expressed and assures higher levels of radioactivity. PMID- 2555760 TI - Characteristics of the cellular response of the rat conjunctiva to topically applied leukotriene B4. AB - Cellular events in the rat conjunctiva were studied at 1, 6 and 24 h following the topical application of leukotriene B4. A significant increase was noted in the number of eosinophils (p less than 0.01) and neutrophils (p less than 0.05) at 1 h and only in eosinophils at 6 h after leukotriene B4 application at the 2 micrograms dose. At the 200-ng dose, the eosinophil count was significantly elevated at 1 h only, and the number of neutrophils was not significantly changed at any time point. No significant increase in cell counts was observed at the 20 ng dose. In conclusion, certain doses of topically applied leukotriene B4 significantly increased the number of eosinophils and neutrophils in the rat conjunctiva. PMID- 2555761 TI - Herpes simplex keratitis in patients with acquired immune deficiency syndrome. AB - Acquired immune deficiency syndrome (AIDS) is associated with a wide spectrum of systemic and ocular infectious diseases. Little information is known about herpes simplex virus type 1 (HSV-1) keratoconjunctivitis in association with AIDS. The authors present six cases of recurrent HSV keratitis occurring in AIDS patients. Features of the herpetic keratitis in these patients included unilateral dendritic or geographic epithelial keratopathy; predilection for peripheral versus central corneal involvement; one to three recurrences per patient over a mean observation period of 17 months, with a median dendrite-free interval of 7 months; and a moderately prolonged clinical course with a median healing time of 3 weeks using topical antiviral therapy. Only one of six cases had stromal infiltrative involvement. These cases raise the question of whether the immunologic abnormalities associated with AIDS may affect the clinical characteristics and course of HSV keratitis. PMID- 2555762 TI - Changes in the superoxide anion generating capacity and respiratory burst enzymes of peripheral blood monocytes of monkeys during acute Plasmodium knowlesi infection. AB - The superoxide anion generation profile of peripheral blood monocytes of rhesus monkeys was investigated during the different stages of an acute Plasmodium knowlesi infection. An initial increase in superoxide anion was followed by a significant decline (P less than 0.001), paralleled by a drop in NADPH oxidase activity; there was no alteration in the activity of the hexose monophosphate shunt enzymes. This lowered activity of the NADPH oxidase, with the resulting decreased O2 generation, might be responsible for the failure of the animals to control the parasitaemia; as a result they succumbed to the infection. PMID- 2555763 TI - Role of mononuclear phagocytes in elimination of Plasmodium chabaudi AS infection. AB - The role of mononuclear phagocytes in acquired immunity resulting in the intraerythrocytic destruction and elimination of malarial parasites was investigated in the murine model of infection with Plasmodium chabaudi AS. Mice were treated 1 day before or 6 days after infection with agents which either result in augmentation or activation of the non-specific, microbicidal effector function of mononuclear phagocytes or in depletion of cells of this lineage. To examine the effect of agents which activate mononuclear phagocytes. A/J mice, which are susceptible to P. chabaudi AS and exhibit fulminant parasitaemia and death within 10 days of intraperitoneal infection with 10(6) P-RBC, were treated intravenously with muramyl dipeptide (MDP) or liposome-encapsulated MDP-glycerol dipalmitate (MDP-GDP). Treatment administered 1 day before infection was ineffective. Treatment on day 6 post-infection with liposome-encapsulated MDP-GDP (1 microgram) resulted in a significant decrease in parasitaemia on day 8 and survival, while treatment with free MDP (100 micrograms) resulted only in a significant decrease in parasitaemia. To examine the effect of depletion of mononuclear phagocytes, C57BL/6 mice, which are resistant to P. chabaudi AS infection and eliminate the parasite by 4 weeks, were treated intravenously with 3 mg silica. Silica administered 1 day before or 6 days post-infection abrogated resistance resulting in a delay in elimination of the parasite and host mortality. Treatment on day 6 was more effective, with death by day 13 post infection of 70% of the normally resistant C57BL/6 mice which exhibited fulminant parasitaemia levels. These results thus provide in-vivo evidence that mononuclear phagocytes play a critical role in the elimination of infection with the murine malaria species P. chabaudi AS. Furthermore, these results suggest that the time of administration of agents which alter mononuclear phagocyte function may be important in determining their effect on host antimalarial defences. PMID- 2555764 TI - Inhibition of lipolysis by agents acting via adenylate cyclase in fat cells from infants and adults. AB - The in vitro lipolytic effect of catecholamines is poor during infancy because of enhanced alpha 2-adrenoceptor activity. The mechanisms behind this were investigated in isolated fat cells obtained from 1- to 4-mo-old infants and from adults. The cells were incubated with agents that inhibit lipolysis through distinct receptors coupled to adenylate cyclase via the inhibitory GTP binding coupling protein, Gi. The sensitivity to the alpha 2-adrenoceptor agonist clonidine was 14 times higher in the infant group as compared to the adults, whereas that to an adenosine analogue was 14 times lower. The sensitivities to prostaglandin E2 and nicotinic acid were similar in both age groups. Preincubation of the adipocytes with pertussis toxin abolished the antilipolytic effects of all agents. The density of alpha 2-adrenoceptor binding sites determined with [3H]yohimbine was increased by about 25% in the infants. In conclusion, the antilipolytic sensitivity of adenosine and alpha 2-adrenoceptors develops separately and may play different roles in the regulation of lipolysis in man. Furthermore, the enhanced alpha 2-adrenoceptor sensitivity during infancy seems at least in part to be due to an increase in the number of receptors. PMID- 2555765 TI - Status of varicella vaccine for healthy children. PMID- 2555766 TI - [Evaluation of functional correlations of intracellular defense factors of neutrophils in children during postnatal ontogenesis]. AB - A total of 310 children aged 1 month to 15 years were examined for functional interrelations between components of the intracellular system of blood neutrophil defence based on the use of a discrete-dynamic analysis. The ontogenetic characteristics and regularities in the development of functional relations of intracellular defence factors were established. The periods of physiologic deficiency in children aged 1 month to 2 years and in those aged 2 to 15 years were defined. A screening scale was suggested for tracing the function of blood neutrophils and for identifying unbalances of intracellular defence factors in the course of the ontogenetic development of children. PMID- 2555767 TI - [Characteristics of antiviral immunity in children with the atopic form of bronchial asthma]. AB - In atopic bronchial asthma, an earlier "encounter" of children with viruses that cause acute respiratory diseases was established as was mass infection of asthma children with respiratory viruses. Disorders of antiviral immunity in children suffering from atopic asthma primarily manifested themselves in overproduction of antibodies among viruses. The leukocyte migration inhibition test has demonstrated that atopic asthma children are sensitized with respiratory viruses. Sensitization to viruses rises with age whereas the presence of the foci of chronic infection promotes ever higher sensitization of the body with respiratory viruses, bringing about inhibition of leukocyte function. PMID- 2555768 TI - [Evaluation of fast MR imaging with suspended respiration in hepatic tumors]. AB - The purpose of this study is to evaluate the possibility of qualitative diagnosis in hepatic tumors by fast magnetic resonance (MR) imaging with suspended respiration using partial flip angle and gradient echo technique at 0.5 T. Fast MR imaging does not replace conventional spin-echo procedures, but is complementary to it. For the analysis of contrast as a function of flip angle, 32 hepatocellular carcinomas (HCCs) of nodular type and 11 hemangiomas were examined with flip angles of 20, 40 and 60 degrees on sagittal images. In general, the lesions showed relatively high and low intensities on the images with flip angles of 20 and 60 degrees, respectively. On the images with flip angle of 40 degrees, signal-to-noise (S/N) ratio was higher, but contrast between tumor and liver was lesser than with that of other angles. The change of contrast-to-noise (C/N) ratio between the flip angles of 20 and 60 degrees in hemangiomas was larger than that in HCCs, significantly. It was useful for evaluation of lesions to observe the change of C/N ratio, and it was necessary for detection of lesions to obtain the images with at least three flip angles. For dynamic MR imaging, 18 HCCs including 5 cases after transcatheter chemoembolization (TCE) and 5 hemangiomas were examined with flip angle of 40 degrees. With employment of Gd-DTPA, S/N ratio and contrast were improved in many cases, and hemodynamics of tumors was able to be observed. It was suggested that dynamic MR imaging was useful especially in evaluation of efficacy of TCE using lipiodol. PMID- 2555769 TI - [The study on radiotherapy for hepatocellular carcinoma]. AB - The effects of radiotherapy on hepatocellular carcinoma (HCC) were studied clinically. Involved-field radiotherapy was employed in the treatment of 27 cases with HCC (irradiation of primary tumor in 12 cases and tumor embolism in 15 cases). Adverse effects of radiation on hepatic functions were minimum, as fluctuations in choline-esterase, prothrombin time and T-bilirubin were insignificant. Changes in ICGR-15 were assessed from the aspects of radiation dose and treatment field. Aggravation was minimum when Time-Dose-Fractionation factor (TDF) was 80 and the involved filed was not more than 8 x 8 cm, but moderate pathologic changes were noted when these levels were exceeded. Clinical effect of the radiotherapy was monitored by medical imagings was remarkable, as tumor regression was evident in 82% of the primary tumors and in 92% of the tumor emboli irradiated. Histologically, improvement of at least clinical stage IIA as described by Ohboshi and Shimosato were observed in 88% of the primary tumors and 80% of tumor emboli treated. When TDF exceeded 80, improvements observed were at least IIA in all cases treated. From these results, it is clear that a TDF80 or higher dose is required for the effective radiation treatment of HCC, and that a treatment field of not more than 8 x 8 cm is safe for the TDF80 dose. As for prognosis of the cases treated with radiotherapy, significant prolongation of the survival period was achieved in the cases with nonresected HCC involved tumor emboli. Thus, radiotherapy for HCC is effective not only for the primary tumor but for tumor embolism as well, and this is a particularly useful modality for the latter. PMID- 2555770 TI - Long term anticoagulation with low molecular weight heparin in outpatients with side effects on oral anticoagulants. AB - A total of 70 outpatients with previous severe haemorrhages and other side effects on conventional oral anticoagulants given for prophylaxis of tromboembolism, were treated with low molecular weight (LMW) heparin fraction Kabi 2165. Anticoagulation was necessary in all patients because of recurrent venous thromboembolism (n = 39), artificial heart valve replacement (n = 12), atrial fibrillation with peripheral embolism (n = 10), and cardiomyopathy (n = 9). LMW heparin was injected sc at doses ranging from 2,500 to 15,000 antifactor Xa (aXa) units once daily by the patients themselves. The dose was adjusted on the basis of body weight, of bleeding risk, and risk of developing thromboembolism. Five of 70 patients with poor compliance, 3 experienced non fatal embolism during the treatment period. Two of 65 patients with good compliance experienced repeat embolism. No fatal embolism occurred. One major episode of gastrointestinal bleeding occurred in a patient with an undetected colon carcinoma. Nine minor hemorrhages were observed in all patients. The present experience suggests that LMW heparin may be used safely and effectively as an alternative anticoagulant in patients who have experienced bleeding and other complications with oral anticoagulants or conventional heparin. PMID- 2555772 TI - In vitro preferential topoisomerization of bent DNA. AB - The steps of the topoisomerization reaction by calf thymus DNA topoisomerase I on a DNA domain containing an intrinsically bent DNA sequence have been analyzed. High preferentiality of binding, cleavage and topoisomerization on the bent segment relative to the rest of the DNA domain was observed. These studies show the importance of the local DNA conformation in the reaction of eukaryotic DNA topoisomerase I and the interest of the use of this enzyme as a tool for the analysis of DNA conformation and DNA dynamics. PMID- 2555773 TI - Interaction of topoisomerase 1 with the transcribed region of the Drosophila HSP 70 heat shock gene. AB - Topoisomerase I cleavage sites have been mapped in vivo on the Hsp70 heat shock gene of Drosophila melanogaster cells using the drug camptothecin. Topoisomerase I cleavage was only observed when the Hsp70 gene was transcriptionally active. Site-specific single-strand DNA cleavage by topoisomerase I was confined to the transcribed region of the Hsp70 gene and occurred on both the transcribed and nontranscribed DNA strands. A number of the single-strand breaks on the complementary DNA strands occurred in close proximity giving rise to double stranded DNA breaks. Inhibition of heat-induced Hsp70 transcription by either Actinomycin D (Act D) or 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) inhibited topoisomerase I cleavage except at the 5' and to a lesser extent the 3' end of the gene. Camptothecin (100 microM) inhibited transcription of the Hsp70 gene greater than 95%. These results suggest that topoisomerase I is intimately associated with and has an integral part in Hsp70 gene transcription. PMID- 2555771 TI - Viral proteins containing the purine NTP-binding sequence pattern. AB - A compilation is presented of viral proteins containing the NTP-binding sequence pattern, and criteria are suggested for assessment of the functional significance of the occurrence of this pattern in protein sequences. It is shown that the distribution of NTP-binding pattern-containing proteins through the viral kingdom is strongly non-random. Sequence comparisons led to delineation of several families of these proteins, some of which could be brought together into superfamilies including also cellular proteins. The available biochemical evidence is compatible with the proposal that viral proteins in which the NTP binding pattern is evolutionarily conserved might all be NTPases involved in: i) duplex unwinding during DNA and RNA replication, transcription, recombination and repair, and possibly mRNA translation; ii) DNA packaging, and iii) dNTP generation. PMID- 2555774 TI - The basis for camptothecin enhancement of DNA breakage by eukaryotic topoisomerase I. AB - The eukaryotic topoisomerase I (topo I) is the target of the cytotoxic alkaloid camptothecin (CTT). In vitro, CTT enhances the breakage of DNA by topo I when the reaction is stopped with detergent. Although breakage at some sites is enhanced to a great extent while breakage at others is enhanced only minimally, CTT does not significantly change the breakage specificity of topo I in vitro. It has been suggested that CTT acts by slowing the reclosure step of the nicking-closing reaction. To test this hypothesis, we have measured the rate of reclosure for different break sites in the presence of CTT after adding 0.5 M NaCl to a standard low salt reaction. In support of the hypothesis, we find that topo I mediated DNA breakage is enhanced the greatest at those sites where closure of the break is the slowest. These results suggest a mechanism for the toxicity of CTT in vivo. PMID- 2555775 TI - The different competitive abilities of viral TAATGARAT elements and cellular octamer motifs, mediate the induction of viral immediate-early genes and the repression of the histone H2B gene in herpes simplex virus infected cells. AB - An HSV virion component, Vmw65, interacts with cellular transcription factors to transactivate TAATGARAT-containing viral genes and some cellular genes containing the related octamer element. We show that the octamer-containing histone H2B promoter can be trans-activated by transfection of Vmw65 but not by viral infection. The induction of H2B transcription by Vmw65 can be abolished by co transfection of excess amounts of either a TAATGARAT element or a Vmw65 responsive octamer element. This effect cannot be overcome by addition of increasing amounts of Vmw65. The H2B promoter and TAATGARAT-containing viral promoters therefore compete for limiting cellular factors required for induction by Vmw65 resulting in repression of the H2B gene during lytic infection. The competitive effect of TAATGARAT elements on the H2B gene is not observed in the absence of Vmw65, but can be produced in the presence of a truncated form of Vmw65 lacking the acidic tail required for transcriptional activation. Hence a domain of Vmw65 distinct from that involved in transcriptional induction interacts with cellular octamer binding proteins favouring binding to the TAATGARAT motif. PMID- 2555777 TI - Nucleotide sequence of the blaZ gene of the Staphylococcus aureus beta-lactamase transposon Tn4002. PMID- 2555776 TI - Functional analysis of the long terminal repeats of Drosophila 1731 retrotransposon: promoter function and steroid regulation. AB - 1731 is a Drosophila retrotransposon whose transcripts decrease in Drosophila cells after treatment by the steroid hormone 20-hydroxyecdysone (20-OH). Several constructions have been made where the bacterial chloramphenicol acetyltransferase (CAT) gene is put under the control of either the 5' or the 3' long terminal repeats (LTRs) of 1731. CAT activity assays in transfected Drosophila cells show that either the 5' or the 3'LTR constitutes a unidirectional promoter. Analysis of partially deleted LTR suggests the presence of so-called silencer and activator regions in these LTRs. Moreover, the first 260 bp of the LTR are sufficient to provoke 20-OH inhibition whereas the first 58 bp are necessary for hormonal responsiveness. These 58 bp contain sequences showing similarities with the targets of trans-acting factors such as Octal-c and NFkB. PMID- 2555778 TI - Complete cDNA sequence of rat phosphoribosylpyrophosphate synthetase subunit II (PRS II). PMID- 2555779 TI - Complete cDNA sequence of rat phosphoribosylpyrophosphate synthetase subunit I (PRS I). PMID- 2555780 TI - Two closely spaced promoters are equally activated by a remote enhancer: evidence against a scanning model for enhancer action. AB - To explain the activation of transcription by a remote enhancer, two models are most often considered, namely looping and scanning. A scanning model, also referred to as 'polymerase entry site' model predicts that for two adjacent promoters the one proximal to an enhancer would be preferentially activated. Preferential activation of the proximal promoter in a tandem promoter arrangement has been found before in several laboratories, including our own, but for technical reasons the data were inconclusive with regards to the enhancer mechanism. In the work presented here, we readdress the question of preferential promoter activation by an enhancer using a more clearly defined system. Two identical promoters were kept closeby in a divergent, or directly repeated orientation. The SV40 enhancer was placed at a great distance on one or the other side of the two promoters, to see whether the enhancer position influenced the relative efficiency of the two promoters in transfected cells. Our finding that the promoter usage is virtually unaffected by the enhancer position does not favor a scanning model, but is compatible with a looping model of enhancer action. PMID- 2555781 TI - Sea urchin egg mitochondrial DNA contains a short displacement loop (D-loop) in the replication origin region. AB - Based on solution hybridization using single-stranded probes, native mitochondrial DNA extracted from sea urchin eggs contains a displacement-loop (D loop) of approximately 70-80 nt. This maps to the single extended unassigned sequence of the genome, between the genes for tRNA(thr) and tRNA(pro), which also appears to contain the origin of first-strand replication. The D-loop commences at or close to a site of supercoil-dependent S1 nuclease hypersensitivity, adjacent to a run of 20 consecutive C residues, terminates near to the boundary of tRNA(thr), and appears to be composed at least partly of RNA, based on the sensitivity of the assays to RNase H. These experiments imply that the mechanisms of replication initiation in sea urchin and vertebrate mtDNAs are very similar, and suggest that the developmental restriction on mtDNA synthesis in eggs and embryos is maintained at the level of D-loop extension. PMID- 2555782 TI - Transcriptional mapping and nucleotide sequence of a vaccinia virus gene encoding a polypeptide with extensive homology to DNA ligases. AB - Nucleotide sequencing of the vaccinia virus SalI F DNA fragment identified an open reading frame of 552 amino acids encoding a protein of 63.3 kDa. The deduced amino acid sequence shares 30% identity with S. pombe and S. cerevisiae DNA ligases, with homology strongest near the carboxy terminus and around the lysine residue required for ligase-adenylate formation. Prokaryotic DNA ligases are poorly related to the vaccinia sequence. The initiation codon of the ORF forms part of a late transcriptional initiation sequence TAAATG and is preceded by two overlapping early transcriptional termination signals, TTTTTTTAT. Nonetheless, RNA mapping showed that the ligase gene is transcribed early during infection and the 5' end of the mRNA maps to the TAAATG motif. The possible roles of a DNA ligase in vaccinia virus DNA replication and recombination are discussed. PMID- 2555783 TI - Sequence and characteristics of IS900, an insertion element identified in a human Crohn's disease isolate of Mycobacterium paratuberculosis. AB - The complete sequence of an insertion element IS900 in Mycobacterium paratuberculosis is reported. This is the first characterised example of a mycobacterial insertion element. IS900 consists of 1451bp of which 66% is G + C. It lacks terminal inverted and direct repeats, characteristic of Escherichia coli insertion elements but shows a degree of target sequence specificity. A single open reading frame (ORF 1197) coding for 399 amino acids is predicted. This amino acid sequence, and to a lesser extent the nucleotide sequence, show significant homologies to IS110, an insertion element of Streptomyces coelicolor A3(2). It is proposed that IS900, IS110, and similar insertion elements recently identified in disease isolates of Mycobacterium avium are members of a phylogenetically related family. IS900 will provide highly specific markers for the precise identification of Mycobacterium paratuberculosis, useful in defining its relationship to animal and human diseases. PMID- 2555785 TI - Repression of human cytomegalovirus gene expression associated with a novel immediate early regulatory region binding factor. AB - A major determinant controlling reactivation of persistent viruses is likely to be the level of cellular factors which regulate virus transcription. Human cytomegalovirus (the largest human herpesvirus) does not replicate in human teratocarcinoma (T2) cells due to a block in transcription of immediate early (IE) gene expression, but these cells become permissive upon retinoic acid induced differentiation. We have analysed changes in DNA binding factors to the major IE promoter/regulatory region of HCMV that occur during differentiation of T2 cells to a permissive phenotype. We show that undifferentiated T2 cells contain a specific nuclear factor that binds to a far upstream region of the major IE regulatory region. Differentiation of T2 cells is associated with a major decrease in this factor and deletion of its specific binding site from IE expression vectors also results in increased levels of expression in undifferentiated cells. Consequently, this novel factor present in undifferentiated cells is a candidate for a differentiation specific negative regulator of HCMV IE gene expression which binds to an element upstream of the major IE enhancer. PMID- 2555784 TI - Mercury-induced transitions between right-handed and putative left-handed forms of poly[d(A-T).d(A-T)] and poly[d(G-C).d(G-C)]. AB - Poly[d(A-T).d(A-T)] and poly[d(G-C).d(G-C)], each dissolved in 0.1 M NaClO4, 5 mM cacodylic acid buffer, pH 6.8, experience inversion of their circular dichroism (CD) spectrum subsequent to the addition of Hg(ClO4)2. Let r identical to [Hg(ClO4)2]added/[DNA-P]. The spectrum of the right-handed form of poly[d(A T).d(A-T)] turns into that of a seemingly left-handed structure at r greater than or equal to 0.05 while a similar transition is noted with poly[d(G-C).(G-C)] at r greater than or equal to 0.12. The spectral changes are highly cooperative in the long-wavelength region above 250 nm. At r = 1.0, the spectra of the two polymers are more or less mirror images of their CD at r = 0. While most CD bands experience red-shifts upon the addition of Hg(ClO4)2, there are some that are blue-shifted. The CD changes are totally reversible when Hg(II) is removed from the nucleic acids by the addition of a strong complexing agent such as NaCN. This demonstrates that mercury keeps all base pairs in register. PMID- 2555786 TI - Identification of multiple cis-elements and trans-acting factors involved in the induced expression of human IL-2 gene. AB - Regulated expression of interleukin-2 (IL-2) gene constitutes an essential part in the clonal proliferation of activated T lymphocytes (T-cells). In order to gain insight on the mechanism(s) of the IL-2 gene induction, deletions were introduced in the human IL-2 gene 5'-flanking region and the mitogen inducibility of each deletion mutant was examined in cultured T cell lines. At least four functional regions were identified, they contain potential binding sites for several transcription factors including one NF-kappa B and two octamer binding factor sites. Whereas each of the functional regions (or elements) is required for the maximal induction of the IL-2 gene by mitogen, one such region was found to be dispensable for activation by the HTLV-1-encoded trans-activator, tax-1. Furthermore, the potent immunosuppressive agent, cyclosporin A was found to inhibit the gene induction by mitogen, but not by tax-1. PMID- 2555787 TI - RNase H cleavage of RNA hybridized to oligonucleotides containing methylphosphonate, phosphorothioate and phosphodiester bonds. AB - Three types of 14-mer oligonucleotides were hybridized to human beta-globin pre mRNA and the resultant duplexes were tested for susceptibility to cleavage by RNase H from E. coli or from HeLa cell nuclear extract. The oligonucleotides contained normal deoxynucleotides, phosphorothioate analogs alternating with normal deoxynucleotides, or one to six methylphosphonate deoxynucleosides. Duplexes formed with deoxyoligonucleotides or phosphorothioate analogs were susceptible to cleavage by RNase H from both sources, whereas a duplex formed with an oligonucleotide containing six methylphosphonate deoxynucleosides alternating with normal deoxynucleotides was resistant. Susceptibility to cleavage by RNase H increased parallel to a reduction in the number of methylphosphonate residues in the oligonucleotide. Stability of the oligonucleotides in the nuclear extract from HeLa cells was also tested. Whereas deoxyoligonucleotides were rapidly degraded, oligonucleotides containing alternating methylphosphonate residues remained unchanged after 70 minutes of incubation. Other oligonucleotides exhibited intermediate stability. PMID- 2555789 TI - Transcriptional activation of cKi-ras proto-oncogene resulting from retroviral promoter insertion. AB - Enhanced expression of the cKi-ras proto-oncogene in a bone marrow-derived mouse cell line, 416B, has been shown to be associated with the integration of Friend viral DNA into the cellular gene. Here we report the results of experiments designed to clarify the molecular mechanism responsible for the cKi-ras overexpression. Based on primer extension analyses and DNA sequencing of cKi-ras cDNA clones, we have obtained evidence that the 416B cells contain viral-host chimaeric transcripts that initiate within the 3' long terminal repeat (LTR) of the integrated provirus. Processing of the transcripts from the rearranged cKi ras gene includes an unexpected splicing event associated with the fortuitous creation of a cryptic donor splice site at the junction between the proviral and cellular DNA sequences. These data demonstrate that enhanced cKi-ras expression in the 416B cells results from a retroviral promoter insertion mechanism of transcriptional activation. PMID- 2555790 TI - Sequence-specific functions of the early palindrome domain within the SV40 core origin of replication. AB - The early palindrome domain within the SV40 core origin of replication is essential for the initiation of replication. Studies with single point mutants in this region suggested that the early palindrome domain does not function as a cruciform structure, but may be involved in the initiation of SV40 DNA replication in a sequence-specific manner. Two mutants, base-substituted at a primase initiation site nucleotide 5214, showed dramatic decreases in DNA replication in monkey cells. Despite earlier reports to the contrary, disruption of the cruciform configuration or polypyrimidine tract does not invariably lead to lack of replication function, as some mutants unable to form this structure replicate normally. Gel retention assays and DNase I footprinting with the nuclear proteins of monkey cells showed that the 5'GAGGC3' pentanucleotide repeats on either side of early palindrome domain interact with monkey nuclear protein. The early palindrome domain may affect the interaction of SV40 DNA with nuclear protein, and participate in SV40 DNA replication. PMID- 2555788 TI - Aphidicolin resistance in herpes simplex virus type I reveals features of the DNA polymerase dNTP binding site. AB - We describe the mapping and sequencing of mutations within the DNA polymerase gene of herpes simplex virus type 1 which confer resistance to aphidicolin, a DNA polymerase inhibitor. The mutations occur near two regions which are highly conserved among DNA polymerases related to the herpes simplex enzyme. They also occur near other herpes simplex mutations which affect the interactions between the polymerase and deoxyribonucleoside triphosphate substrates. Consequently, we argue in favor of the idea that the aphidicolin binding site overlaps the substrate binding site and that the near-by conserved regions are functionally required for substrate binding. Our mutants also exhibit abnormal sensitivity to another DNA polymerase inhibitor, phosphonoacetic acid. This drug is thought to bind as an analogue of pyrophosphate. A second-site mutation which suppresses the hypersensitivity of one mutant to phosphonoacetic acid (but not its aphidicolin resistance) is described. This second mutation may represent a new class of mutations, which specifically affects pyrophosphate, but not substrate, binding. PMID- 2555791 TI - The analysis of the human hemopexin promoter defines a new class of liver specific genes. AB - Hemopexin (Hpx) is a plasma glycoprotein which is expressed only in the liver. It is synthesized at a lower rate in the fetal liver than in the adult, and its level increases during acute infections. As shown here, a fragment of the human hemopexin promoter spanning from positions -130 to +22 relative to the cap site is sufficient to direct cell-specific transcription of a reporter gene. Within this segment a short sequence, located between positions -120 and -104, is responsible for this effect. This positive cis-acting element, the Hpx A site, interacts with a family of nuclear proteins, some of which are present only in hepatoma cells. The potential meaning of these complex DNA-protein interactions and the homology with elements present on the promoter of other liver-specific and acute phase genes are discussed. PMID- 2555793 TI - Two classes of observed frequency for rare-cutter sites in CpG islands. PMID- 2555792 TI - A negative regulatory element in the promoter of the human alpha 1-antitrypsin gene. AB - A "minimal tissue-specific element" of the human alpha 1-antitrypsin (alpha 1-AT) gene promoter, located in the -137/-37 region, is able to drive hepatoma-specific transcription from the heterologous SV40 promoter. Here we show that, in HeLa cells, this element is inactivated by a negative regulatory mechanism. This negative control can be overcome by deletion or mutation of two specific DNA domains. PMID- 2555794 TI - pBluescript II: gene mapping vectors. PMID- 2555795 TI - Inhibition of restriction endonuclease hydrolysis by phosphorothioate-containing DNA. PMID- 2555796 TI - A rapid and quantitative microtiter assay for eukaryotic topoisomerase II. PMID- 2555797 TI - Effect of VIP on the respiratory burst in human monocytes ex vivo during prolonged strain and energy deficiency. AB - VIP-stimulated cyclic AMP production and VIP effect on the production of reactive oxygen compounds in human monocytes activated by serum opsonized zymosan (respiratory burst) were studied during a ranger training course lasting for five days with almost continuous physical activity, and deficiency of sleep and energy. Respiratory burst was inhibited and cyclic AMP production was stimulated by VIP on all days. Maximum cyclic AMP production stimulated by VIP (0.1 microM) on the day of control was 148.6% of basal, and 255.3%, 213.8%, 218.9% and 198.7% on Days 1, 2, 3 and 5. Maximum inhibition was observed 20 min after addition of the peptide on the day of control, after 5 min on Days 1, 2 and 3, and after 10 min on Day 5. Inhibition at the 5-min time point was 33.1% on the day of control, and 34.7%, 53.6%, 53.3% and 36.2% on the different days during the training course. The observed increment in VIP effect adds to prior reported data about increased VIP secretion during the training course, and may indicate enhanced physiological significance of VIP during stress. PMID- 2555798 TI - TRH stimulates the release of POMC-derived peptides from goldfish melanotropes. AB - The release of immunoreactive (ir) alpha-MSH and ir ACTH from goldfish (Carassius auratus) melanotropes was investigated using superfused isolated dispersed neurointermediate lobe cell columns. Stimulation of neurointermediate lobe cell columns with pulses of TRH evoked dose-dependent increases in the concomitant release of ir alpha-MSH and ir ACTH. Reversed-phase high performance liquid chromatography (RP-HPLC) was used to characterize the alpha-MSH and ACTH immunoreactivities released from a neurointermediate cell column under spontaneous release conditions. Six peaks of ir alpha-MSH were revealed. Three of these peaks were identified as des-acetyl alpha-MSH, mono-acetyl alpha-MSH and di acetyl alpha-MSH. Seven peaks of ir ACTH were revealed. Four of these peaks were tentatively identified as ACTH variants. These studies suggest that TRH stimulates the release of peptide hormones from teleost melanotropes and that the goldfish neurointermediate lobe in vitro releases numerous peptides derived from POMC. PMID- 2555799 TI - Relationship between DNA fragmentation and apoptosis in the programmed cell death in the rat prostate following castration. AB - Previous studies have demonstrated that the rapid involution of the rat ventral prostate following castration involves the death of the androgen-dependent epithelial cells present within the gland and that this death is the result of a series of discrete biochemical steps. The degradation of genomic DNA into nucleosomal-sized fragments is an early event in this process and is catalyzed by calcium magnesium-dependent endonuclease activity. The morphologic correlation of the involution process involves a series of structural changes which are collectively referred to as apoptosis. The apoptotic process describes the earliest apparent signs of morphologic change exhibited by the dying cells through their eventual complete destruction and deletion from the tissue. The temporal relationship between these recently described biochemical events and the morphologic changes of the apoptotic process were compared in the present study, in order to test the cause versus effect nature of DNA fragmentation in the programmed death of androgen dependent prostatic cells following castration. These studies demonstrated that the early elevation of the Ca+2 Mg+2-dependent endonuclease activity and the fragmentation of DNA into nucleosomal oligomers occurs within prostatic glandular epithelial cells and probably does not involve the direct participation of extraprostatic cells which may subsequently migrate into the gland. Once the DNA is initially cleaved into the nucleosomal oligomers, the subsequent participation of lysosomal enzymes act in a less restricted fashion to degrade both the nucleosomal DNA as well as the cytoplasmic elements and the cell becomes morphologically apoptotic. As the elevations in Ca+2 Mg+2 dependent endonuclease activity and DNA fragmentation are initiated at a time well before the cell is morphologically dead, as defined by apoptosis, these changes in DNA metabolism must not be the consequences of cell death but instead are early causal events in an active process of programmed cell death. PMID- 2555800 TI - Cushing's syndrome. How to pinpoint and treat the underlying cause. AB - In addition to prolonged glucocorticoid therapy (not discussed here), at least five other conditions cause Cushing's syndrome. They are excessive corticotropin secretion by the pituitary gland (which results in Cushing's disease), ectopic production of corticotropin by malignant nonpituitary tumors, benign adrenal adenoma, adrenal carcinoma, and primary adrenocortical nodular dysplasia. Each can be distinguished by a specific pathophysiologic process that triggers the adrenal glands to overproduce glucocorticoids. At present, diagnosis of Cushing's syndrome or disease relies heavily on the dexamethasone (Decadron, Hexadrol) suppression test. After diagnosis, other studies, including computed tomography, magnetic resonance imaging, and corticotropin radioimmunoassay, can be used to localize the site of the lesion. Treatment, of course, depends on the underlying cause. PMID- 2555801 TI - Genetic variation of peptidase and pyrophosphatase in the chicken. AB - Differences in electrophoretic mobilities of two chicken liver, kidney, and spleen enzymes have been demonstrated. A peptidase cleaving the dipeptide L leucyl-alanine was found to be under the control of a single locus, Pep-1, in crosses between two breeds of chicken, White Leghorn (WL) and Rhode Island Red (RIR). Three alleles, a, b, and c were segregating in the WL breed but only two of these, a and c, seemed to be present in the RIR stock. The other enzyme investigated here was pyrophosphatase, and was shown to be under the control of one locus, Pyp, with two alleles, a and b. The two alleles had similar frequencies in the RIR breed, whereas in the WL breed, the a allele was more frequent. PMID- 2555802 TI - Anderson Fabry disease--an identifiable disorder. PMID- 2555803 TI - Psychomotor, respiratory and neuroendocrinological effects of a mu-opioid receptor agonist (oxycodone) in healthy volunteers. AB - Psychomotor performance related to driving and occupational skills was measured double-blind and cross-over in 9 healthy volunteers before and 1.5, 3 and 4.5 hr after intramuscular injection of oxycodone (0.13 mg/kg), oral diphenhydramine (100 mg) and placebo. The effects of oxycodone on performance peaked at 1.5 hr when it prolonged reaction time and impaired vigilance, attention, body balance and coordination of extraocular muscles. The subjects assessed themselves mentally slow, muzzy and impaired by performance on visual analogue scales still 3 hr after injection. Critical flicker discrimination was impaired and some respiratory depression still present at 4.5 hr after administration. Oxycodone elevated plasma prolactin at 1.5 and 3 hr while growth hormone levels remained unaffected. We conclude that the profile of psychomotor decrement produced by this mu-opioid agonist closely resembles that of agonist-antagonist analgesics. PMID- 2555805 TI - Increased methylation of chloroform extractable products and CTP: cholinephosphate cytidylyltransferase in brain membrane preparations from triethyltin-intoxicated rats. AB - Rats were chronically intoxicated with triethyltin in the drinking water (0.002%) for a period of 15 days. Starting with day 5 of the intoxication period a decrease in the body weight was observed and, in parallel, the development of a cerebral oedema could be followed by measuring white matter density. At the same time, an increase of phosphatidylethanolamine-N-methyltransferase and cholinephosphate cytidylyltransferase activities was noted. This increase might be a compensatory mechanism for counteracting the membrane damages induced by triethyltin. PMID- 2555804 TI - Effect of pinacidil on sympathetic neuroeffector transmission in rabbit blood vessels. AB - The effects of pinacidil on vascular sympathetic neuroeffector transmission were studied. Pinacidil (3 x 10(-7)-3 x 10(-4) M) inhibited the contractions of rabbit isolated pulmonary artery evoked by nerve stimulation. This was the case both in the absence and presence of cocaine plus corticosterone. The inhibition reached a steady state and was reversible. Pinacidil (10(-5) -3 x 10(-4) M), in the absence or presence of cocaine plus corticosterone, markedly enhanced the 3H-overflow evoked by electrical-field stimulation of the artery preloaded with 3H noradrenaline (3H-NA). The enhancement was dependent on frequency (1-30 Hz) in a complex manner. Phentolamine (3 x 10(-6) M), but not rauwolscine (10(-6) M) prevented the enhancement. Pinacidil (10(-4) M) did not antagonize the alpha methylnoradrenaline-induced inhibition of 3H-overflow evoked by stimulation. Pinacidil (10(-5)-3 x 10(-5) M) in a non-competitive manner antagonized the contractions of isolated aorta elicited by noradrenaline (3 x 10(-9)-3 x 10(-5) M), phenylephrine (2 x 10(-8)-3 x 10(-4) M), adrenaline (10(-9)-3 x 10(-5) M), histamine (10(-6) 6 x 10(-4) M), serotonin (10(-8)-10(-4)M), and potassium (10( 3) M). Pinacidil (10(-6)-3 x 10(-4) M) and methacholine (3 x 10(-8)-10(-6) M) relaxed aorta preconstricted with noradrenaline (10(-7) M). The relaxation caused by pinacidil was not dependent on the presence of endothelial cells, while that seen with methacholine was.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555806 TI - A study of the possible mechanisms underlying the convulsant actions of a linear expanded xanthine. AB - The actions of a mixture of the 4- and 9-chloro derivatives of the linear expanded xanthine 5,7-diethyl-2-chloroimidazo[4,5-g]quinazoline-6,8 (5H,7H)-dione (chloro-DCQD) on the isolated olfactory cortex slice of the rat have been investigated. Chloro-DCQD evoked a slowly-developing depolarization which intensified over a drug application period of at least 4 min. A pharmacological investigation of the response showed that it was not mediated by blockade of potassium channels or activation of voltage-gated sodium channels, by a stimulant action at receptors to gamma-aminobutyric acid (GABA), excitatory amino acids or acetylcholine, or by antagonism of adenosine receptors. Chloro-DCQD (2.5 mM) potentiated responses evoked by N-methyl-D-aspartate (NMDA), L-aspartate and L glutamate, probably by overcoming the magnesium ion block of the ion channel of the NMDA receptor complex. Chloro-DCQD (2.5 or 5 mM) also increased pyramidal cell excitability and abolished GABA-mediated postsynaptic inhibition but did not affect the excitability of, or neurotransmitter release from, the terminals of the lateral olfactory tract. Chloro-DCQD competitively antagonized the inhibitory actions of adenosine on the olfactory cortex. These effects are consistent with the reported convulsant actions of chloro-DCQD. PMID- 2555807 TI - Impaired beta-adrenergic mediated cerebral blood flow response in streptozotocin diabetic rats. AB - The influence of intracarotic infusion of isoproterenol on cerebral blood flow, on cerebrovascular CO2-reactivity, and on cerebral glucose metabolism were investigated in streptozotocin diabetic rats. Resting cerebral blood flow in diabetic rats decreased by 26% from a control value of 90 ml/100 g.min. to 66 ml/100 g.min. Intracarotid isoproterenol infusion leads to a significant increase in cerebral blood flow in both groups. The increase in cerebral blood flow in control rats (70%), significantly exceeded that in the streptozotocin diabetic rats (33%). Cerebrovascular reactivity to carbon dioxide was preserved after isoproterenol infusion in both groups and there was no significant difference in the relative cerebral blood flow increase in the two groups. Cerebral glucose metabolism was also unaffected by the isoproterenol infusion. The results indicate that impaired beta-adrenergic mechanisms may play a role for alterations in cerebral blood flow that is seen in diabetes mellitus. PMID- 2555808 TI - Electron spin resonance of eumelanin from hair: photoinduced radicals in solid matrix. AB - KBr matrices appear to be convenient media to reveal the radicals formed on light exposure of eumelanin dispersions. The ESR signal of eumelanin dispersed at low concentration in KBr pellets is analyzed during and after irradiation at various wavelengths. Different types of radicals are observed. R'1- and R1-types of radicals are assigned, respectively, to neutral and deprotonated intrinsic phenoxy radicals of eumelanin. R'1 can be oxidized by oxygen as opposite to R1. R2- and R'2-types are formed in the indolic site. Water favours the conversion of R2, unreactive with oxygen, into R'2 which can be oxidized. R'1 and R2 result of an electron photoejection, respectively, from the phenolic and the indolic site. The R3-type radicals are associated with the band-to-band excitation of eumelanin considered as a semiorganized solid. PMID- 2555809 TI - Electron spin resonance study of photoinduced radicals in various eumelanins in solid matrix. AB - The photoinduced radicals formed in eumelanin from hair at various excitation wavelengths, in KBr matrix, are compared with the photoinduced radicals observed with bovine eye iris and squid ink eumelanins. Qualitative similarities and quantitative differences are observed, specially on exposure at short wavelengths. PMID- 2555810 TI - Phosphorylation of the carotenoid droplet protein p57 by the catalytic subunit of cyclic adenosine monophosphate-dependent protein kinase and the effect of fluoride. AB - We have previously shown that the dispersion and aggregation of carotenoid droplets in goldfish xanthophores are regulated, respectively, by phosphorylation and dephosphorylation of a carotenoid droplet protein p57. There is a basal level of p57 phosphorylation of p57 in unstimulated cells, which is greatly stimulated by adrenocorticotropic hormone (ACTH) or cyclic adenosine monophosphate (cAMP) acting via cAMP-dependent protein kinase. We have also observed that, in permeabilized xanthophores, pigment dispersion can be induced when cAMP is replaced by fluoride. Since p57 has multiple phosphorylation sites, there is the question of whether all p57 phosphorylation is by cAMP-dependent protein kinase or whether phosphorylation by cAMP-independent protein kinase coupled with inhibition of phosphatase activity by fluoride can replace cAMP-dependent protein kinase and that the ability of fluoride to replace cAMP for pigment dispersion in permeabilized cells is probably due to activation of adenylcyclase. We also show that ACTH causes an approximately threefold increase in the level of cAMP in these cells. PMID- 2555811 TI - Epinephrine-induced pigment aggregation in goldfish melanophoroma cells: apparent involvement of an unknown second messenger. AB - Using a goldfish-derived melanized cell line, we attempted to determine the identity of the signal transduction system/second messenger for epinephrine induced aggregation of melanosomes in a goldfish cell line. The results show that the second messenger is unknown. It is not 1) influx of extracellular calcium, 2) release of intracellular stored calcium via the phosphoinositide pathway, 3) cGMP, or 4) decrease of cAMP. These results suggest that there is an unknown second messenger for this activity of epinephrine. PMID- 2555812 TI - [Erythema nodosum associated with parvovirus B19 infection]. PMID- 2555813 TI - [Causes of infectious gastro-enteritis in children in Dakar]. AB - Between February 1983 and May 1988, 1,157 stools from children aged under 15 years presenting with diarrhoea and admitted to a paediatric hospital in Dakar, Senegal, were examined for the presence of bacterial and parasitic agents. We looked for Campylobacter and rotavirus in only 245 and 111 samples respectively. Enterobacteria were detected most frequently (162/264; 61.3 per cent). Among these, we found 92 (34.8 per cent) strains of enteropathogenic E. coli; 40 strains (15.1 per cent) of Salmonella spp. and 30 strains (11.3 per cent) of Shigella spp. V. cholerae was present in only 2 faecal specimens (0,7 per cent). Eighty-one (30.6 per cent) parasitic agents were isolated, including a high proportion (13.6 per cent) of flagellae. Campylobacter jejuni and retroviruses were found in 2 (0.8 percent) and 15 (13.5 percent) stool respectively. The frequency with which these enteropathogenic agents were isolated remains in keeping with the global situation usually observed in Dakar for several years. PMID- 2555814 TI - [The hypothalamus-hypophysis-adrenal system in patients with lactorrhea amenorrhea syndrome]. AB - Function of the hypothalamo-hypophyseal-adrenal system was investigated in 23 patients with different types of the lactorrhea-amenorrhea syndrome. The basal levels and "stress secretion" of ACTH, cortisol and cyclic nucleotides, the circadian rhythm of ACTH and cortisol levels were determined by RIA. Noticeable disorders in the central and peripheral factors of this system and changes in the blood levels of cyclic nucleotides were observed in patients with the lactorrhea amenorrhea syndrome. PMID- 2555815 TI - [Magnetic resonance tomography of the adrenals in the diagnosis of total and partial hypercorticism]. AB - The paper is devoted to the use of a new method of nuclear magnetic resonance tomography (MR tomography) in the diagnosis of diseases of the adrenal glands developing with the syndrome of total and partial hypercorticism. A MR tomograph with resistive magnet (0.234 T) was employed. Altogether 25 patients were examined (21 with Itsenko-Cushing disease of various degrees of severity, 2 patients with Itsenko-Cushing syndrome and 2 patients with aldosteronoma). MR tomography was shown to possess a high informative value permitting the assessment of a type of adrenal changes without irradiation of a patient. In combination with routine methods of examination (hormonal investigation, retropneumoperitoneum, x-ray computerized tomography) MR tomography makes it possible to improve considerably the diagnosis of Itsenko-Cushing disease and to specify tumor sizes and tumor growth type. PMID- 2555816 TI - [A rare case of polyhormonal hypophyseal adenoma]. PMID- 2555818 TI - Detection of human papillomaviruses 16-18 in cervical cells by molecular hybridization: relationship with morphonuclear cell image analyses. AB - Forty-one Feulgen-stained cervical imprint smears were analyzed by means of the SAMBA 200 cell image processor in order to quantitatively score human papillomavirus (HPV) 16-18-induced morphonuclear modifications as assessed by morphometric, densitometric, and textural parameters. Molecular hybridization technology using 16 and 18 type specific genetic probes made it possible to divide our series into three groups: Group 1, containing noninfected smears; Group 2, containing "suspicious", i.e., borderline positive, smears; and Group 3, those related to infected patients. Our results show that nuclei from infected smears are much more hyperchromatic and bigger than those arising from noninfected smears. This quantitative description of HPV 16-18-induced chromatin modifications enabled us to create preliminary data banks which could lead to an objective and reproducible grading of unknown cases. This approach is now being prospectively assessed on a large series of cases because the value of the current study is limited until the data bank is tested against unknown specimens with a broader spectrum of HPV infection. PMID- 2555817 TI - [The neuroendocrine effects of neonatal exposure to an inhibitor of catechol-o methyltransferase and of sex steroids]. AB - The role of catechol-o-methyltransferase (COMT) in functional interrelationship between testosterone (T), catechol estrogens (CE) and catecholamines (CA) during cerebral sex differentiation (CSD) was investigated in experiments on Wistar rats. Sex dimorphism in the level of CA in the rat hypothalamus was revealed on the 10th day of life. Noradrenaline concentration in male rats was significantly higher than in the female rats (p less than 0.05). It was shown that isolated CA accumulation in the hypothalamus of 10-day female rats by means of direct suppression of COMT with tropolon (300 micrograms on the 5th and 7th days of life) was insufficient for masculinization of sex cycling regulation centers. At the same time tropolon administered in a dose of 100 micrograms on the 4th-10th days of life enhanced the sterilizing effect of T administered in a dose of 25 micrograms on the 4th day of life. The development of anovulatory sterility (AS) was observed in 100% of cases. The neonatal effect of 2-hydroxyestradiol-17 beta (2-OH-E2 50 micrograms on the 5th day of life) and tropolon (300 micrograms on the 5th and 7th days of life) was ineffective with relation to AS induction indicating the absence of the inductor role of 2-OH-E2 in CSD. A conclusion is that CSD is a result of combined action of androgens, their metabolites (4 hydroxylated CE isomers) and COMT-mediated CA. PMID- 2555819 TI - An unusual embryonal tumor of the nasal cavity in a neonate--a nasal blastoma? A clinical, histologic, and ultrastructural study. AB - We report the clinicopathological and ultrastructural features of a primitive embryonal tumor which occurred as a polypoid mass in the nasal cavity of a newborn infant. Its primitive-appearing, glycogen-rich, small round cells exhibited epithelial characteristics histologically, immunohistochemically, and ultrastructurally. The tumor was initially diagnosed as embryonal carcinoma because of evidence for endodermal differentiation and lack of specific features of other types of small round cell tumors of childhood. However, it did not have the characteristic features of endodermal sinus tumor nor the anaplasia of the adult type of embryonal carcinoma. It had some resemblance to embryonic somatic tissues and the blastemal component of some of the primitive tumors of childhood. Because of its unique morphological appearance, which has never been reported, and its relatively indolent behavior after chemotherapy and radiotherapy, we believe that this is a distinctive primitive teratoid tumor which may be classified as nasal blastoma. PMID- 2555820 TI - Phagocytosis of particulate activators of the human alternative complement pathway through monocyte beta-glucan receptors. AB - Human monocytes phagocytose particulate activators of the alternative complement pathway through beta-glucan receptors in the absence of opsonins. Recognition of soluble beta-glucans by monocytes selectively inhibits ingestion of particulate activators and has no effect on responses mediated by monocyte receptors for Fc IgG, complement, or fibronectin. The smallest ligand unit recognized by monocyte beta-glucan receptors is an acid-resistant heptaglucoside present in yeast cell walls. Mouse monoclonal anti-beta-glucan antibodies have been prepared, one of which completely neutralizes the inhibitory capacity of the HPLC-purified heptaglucoside. This antibody has been used as immunogen for the preparation of an anti-Id. The pretreatment of monocytes with low concentrations of anti-Id inhibits monocyte ingestion of zymosan particles but not EsIgG, suggesting that this antibody has specificity for monocyte beta-glucan receptors and is a powerful probe for further receptor studies. Other receptors with specificities for carbohydrates are also present on mononuclear phagocytes. Receptors for mannose/fucose and those for galactose have been isolated and cloned. The development of probes, such as structural analogs of the active heptaglucoside and the anti-Id, will bring the beta-glucan receptors to a similar stage of definition. A major factor that adds a considerable degree of difficulty to studies of the beta-glucan receptors and is not shared by the other receptors for carbohydrates is the requirement for structural conformations provided by the alignment of several glucose units rather than the recognition of a hexose residue in, for example, a glycoconjugate. The designation of these receptors as beta-glucan receptors has inadvertently taken us into a new area of nonimmune defense. Animal studies indicate that beta-glucans with 1,3- and/or 1,6-linkages are active pharmacologic agents that rapidly confer protection to a normal host against a variety of biologic insults. The beta-glucan receptors provide a mechanism by which a heightened state of host responsiveness is initiated. PMID- 2555821 TI - Enzymes of the 5-lipoxygenase pathway. PMID- 2555822 TI - Does acidosis contribute to stress-induced ulceration in rat stomachs? AB - The present study examines the involvement of acidosis in stress ulceration in rat stomachs. Cold restraint stress for 2 hr did not affect the blood lactate level; however, it produced respiratory acidosis, as reflected by the depressed respiratory rate which was associated with increased CO2 tension and a lowered blood pH. Severe hemorrhagic ulceration was found in the glandular mucosa. The effects of stress on blood pH and the stomach were reversed by IV infusion of NaHCO3. Infusion of HCl IV decreased the blood pH and HCO-3 level and produced gastric ulceration. It is concluded that respiratory acidosis could be involved in stress ulceration. The metabolic acidosis evoked by HCl also induced gastric damage, but the effect was much less. PMID- 2555823 TI - Antagonism of ethanol-reinforced behavior by the benzodiazepine inverse agonists Ro15-4513 and FG 7142: relation to sucrose reinforcement. AB - The partial inverse benzodiazepine agonist Ro15-4513 has been shown to antagonize many of ethanol's actions, including the reduction of behavior reinforced with ethanol presentation. The studies reported here compared the effects of the Ro compound on sucrose reinforcement alone and concurrently available with ethanol reinforcement. Also, a second inverse agonist, FG 7142, was tested. The result indicated that ethanol reinforcement was more sensitive to the inverse agonists compared to sucrose reinforcement. This was seen as a graded effect upon ethanol responding at doses which failed to have any effect upon sucrose-reinforced behavior. The Ro compound was approximately three times more potent than the FG compound in suppressing ethanol-reinforced responding. Possible explanations for the greater sensitivity of ethanol reinforcement compared to sucrose reinforcement was discussed in terms of ethanol's potential actions at the benzodiazepine-GABA receptor complex. PMID- 2555824 TI - Tripelennamine interactions with the psychotomimetic sigma agonist N allylnormetazocine. AB - The pharmacological effects of individual and combined intravenous doses of the antihistamine tripelennamine and the psychotomimetic sigma benzomorphan opioid derivative, N-allylnormetazocine (NANM), on nociceptive reflexes, autonomic parameters and behavior were assessed in the chronic spinal dog. NANM (1.65 mg/kg, IV) produced antinociception, mydriasis, tachycardia, hyperthermia and behavioral signs of canine delirium. Tripelennamine (1.25 mg/kg, IV) produced antinociception, mydriasis and tachycardia without affecting behavior. The combined effects of the two drugs were additive except for heart rate. However, tripelennamine did not antagonize any of the physiological effects or the signs of canine delirium produced by NANM. The findings are inconsistent with the hypothesis that tripelennamine antagonizes the psychotomimetic NANM-like effects of pentazocine to make pentazocine-tripelennamine combinations (T's and Blues) more desirable as a heroin substitute. PMID- 2555825 TI - R 56865 differentiates between contractile agents with respect to the nifedipine sensitive component in the isolated rat aorta. AB - The interaction of the benzothiazolamine R 56865 with the nifedipine-sensitive component of the serotonin (5-HT)-, angiotensin II (AII)- and arginine vasopressin (AVP)-induced contractions was studied in the isolated rat aorta. Nifedipine caused concentration-dependently (10(-9)-10(-6) mol/l) a slight rightward shift accompanied by a limited depression of the maximum of the concentration-response curves for 5-HT-, AII- and AVP-induced contractions. R 56865 (10(-5) mol/l) antagonized the contraction elicited by 5-HT and AII in a similar manner as nifedipine. The effect of R 56865 on 5-HT- and AII-induced contractions was no longer observed after pretreatment with nifedipine. The AVP induced contraction was not affected by R 56865 (10(-5) mol/l). As shown previously, R 56865 is a weak inhibitor of potential-operated channels but inactive on Ca2+ channels activated by NA. In conclusion, R 56865 does not only differentiate between depolarization and receptor-stimulation, but also between the activation of Ca2+ channels by different types of receptors. We propose that R 56865 may interact with Ca2+ channels at a site which plays a role in their activation. PMID- 2555826 TI - Signal processing in a simple vertebrate photoreceptor system: the teleost pineal organ. AB - The integrating circuitry and efferent pathways for neural signals evoked in the photosensory pineal organ by changes in ambient illumination have been investigated by a multidisciplinary approach. Intrapineal efferent neurons were identified by means of retrograde filling with horseradish peroxidase (HRP). In addition to several types of neurons, photoreceptor cells that emitted axons to the brain via the pineal tract were observed. The presence of several populations of local interneurons (putatively cholinergic, GABAergic and substance P containing) and possible afferent (putatively noradrenergic and peptidergic) central innervations were established by means of immunocytochemistry. The anatomical substrate for processing of neural signals thus delineated, the responses of pineal sensory and neural elements to photic stimulation were investigated by means of intracellular recording. Successful recordings were followed by intracellular injection with HRP or Lucifer Yellow CH, for subsequent light or electron microscopical investigation. The recordings indicate the presence of at least two types of photoreceptor cells, that display morphological and physiological features of both retinal rods and cones. In addition, one type of (sign-conserving) interneuron was identified. The photosensory pineal organ thus possess an integrative neural circuitry that may be involved in the elaboration of neural signals to the brain, and/or in the local control of intrapineal functions, e.g. indoleamine synthesis. PMID- 2555827 TI - Breast cancer in a patient with gynecomastia. AB - A case of breast carcinoma in the midst of florid gynecomastia in a 20-year-old man is reported. Up to now, only two male patients under the age of 21 with breast malignancy have been described in the literature. In contrast, gynecomastia is a more common condition than generally appreciated. The association of gynecomastia, a rather common condition, and cancer of the male breast, a rather uncommon condition, is examined. Mammography is recommended as part of the workup. A new classification of gynecomastia into true, pseudo, and mixed types is suggested. Recommendations for the role of lipoplasty in the treatment of gynecomastia are made. PMID- 2555828 TI - [Determination of fucokinase activity in the blood of schizophrenic patients]. AB - An investigation was conducted into whether dopamine induces an alteration in the fucolysation of glycoproteins, starting from the dopamine hypothesis of schizophrenia and taking the fucokinase activity determined in erythrocytes of schizophrenic patients as parameter. As with patients with "schizoaffective psychosis" and those with manic-depressive disorders, who were likewise examined, it was found that the enzyme activity of schizophrenic patients was no different than that found in the blood of a control group. PMID- 2555829 TI - Effects of dietary polyunsaturated fatty acid on platelet aggregation in rats. AB - In the present study, we changed the fatty acid profile in blood and platelet membranes by dietary manipulation, and examined the effect on platelet aggregation in rats. Fifty-five rats were divided into five groups and fed for 56 days with 1% cholesterol and different types of fatty acid-rich diets: basal, lard, lard + fish oil, soybean oil, and soybean oil + fish oil. a decrease in serum arachidonic acid (20:4, omega-6, AA) and an increase in serum eicosapentaenoic acid (20:5, omega-3, EPA) were found in all experimental dietary groups fed with refined fish oil. Similar changes in the polyunsaturated fatty acids were also found in the platelet membrane phospholipids. Platelet aggregation, quantitated by the slope and height of the aggregation curve induced by different concentrations of ADP in a platelet aggregometer, was inhibited in all groups fed with refined fish oil. This inhibition of platelet aggregation may be related to an increase in the ratio of EPA and AA in the platelet membrane phospholipids after dietary manipulation. The differences in the platelet aggregation and thromboxane B2 (TXB2) concentration between the lard and the lard + fish oil groups were more profound than that between the soybean oil and the soybean oil + fish oil group. However, the malondialdehyde (MDA) concentration revealed no significant differences between the five groups. PMID- 2555831 TI - [Postradiation changes in the activity of membrane-bound nerve tissue enzymes]. AB - A study was made of the effect of various radiation doses (1.75 to 12.25 Gy) on the enzyme activity of Na,K-ATPase system of the microsomal brain fraction of mongrel and Wistar rats. With a similar method of the fraction isolation different response of the activity of this enzyme was registered. Different radiosensitivity of M9-ATPase is responsible for the direction of changes in the Na,K-ATPase activity of the preparations. PMID- 2555830 TI - Feedback modulation of acetylcholine release from motor nerve. PMID- 2555832 TI - [The radiomodifying action of ATP and ADP on membrane-bound enzymes]. AB - A modifying action of ATP and ADP on the activity of some key membrane-bound enzymes of the brain and heart microsomes of rats exposed to 7 Gy radiation has been investigated. The difference in the reactions of energy-dependent enzymes is attributed to the compensatory systems involved at the molecular level. PMID- 2555833 TI - [Quantitative changes in the ultrastructural elements of presynapses exposed to low-intensity laser radiation within the framework of a factorial model]. AB - The ultrastructure elements of presynaptic terminals (PT) in a dorsal horn of cat spinal cord were studied morphometrically in norm and after helium-neon laser irradiation. The disperse computer analysis showed changes in a median terminal radius, the number and localization of synaptic vesicles, and no changes in the shape and length of the plasmalemma profile of the irradiated PT. PMID- 2555834 TI - Nicotinic receptors in the CNS. Their role in synaptic transmission. Proceedings of an international symposium. Uppsala, Sweden, June 19-21, 1988. PMID- 2555835 TI - Diagnosis, occurrence and clinical significance of the human 'candidate' caliciviruses. PMID- 2555836 TI - Potential use of nonpathogenic enteroviruses for control of human disease. AB - Until recently, it has been generally assumed that all human viruses are causative agents of diseases and should be regarded as harmful pathogenic agents that require control measures. In the early 1950s we began to doubt this view. In the course of experiments on virus isolations from feces of normal children, as well as in studies of isolates from animals and from tissue cultures, data accrued which suggested that some conditionally pathogenic and some completely nonpathogenic strains of enteroviruses may provide some benefit to their host by inhibition of pathogenic viruses and by activating nonspecific protective functions of the organism. The novel concept of beneficial viruses was proposed which suggested that the process of co-evolution of the host organism and its associated viral flora led to a specific interaction between them that was beneficial for both. This concept provides a potential approach to the nonspecific prevention of viral diseases by means of the interference between beneficial enteroviruses and pathogenic viruses belonging to different classes. PMID- 2555837 TI - Occurrence and significance of papovaviruses BK and JC in the urine. PMID- 2555838 TI - [Calcium release channels of sarcoplasmic reticulum]. PMID- 2555839 TI - [Progress in studies on stringent regulation (ppGpp)]. PMID- 2555840 TI - [Microtubules and protein phosphorylation]. PMID- 2555841 TI - [Actin regulatory proteins]. PMID- 2555842 TI - [Actin binding proteins from physarum and the organization of the actin cytoskeleton]. PMID- 2555843 TI - [Growth factor-induced changes in the actin architecture]. PMID- 2555844 TI - [Axonal cytoskeleton]. PMID- 2555845 TI - [Abnormal proteins in the brain in Alzheimer's disease]. PMID- 2555846 TI - Diagnosis of coexistent hepatic tumors of different kinds: a challenge from CT and MR imaging. AB - Three cases of multiple, different liver tumors in the same patient were examined with CT and MR imaging. Two patients with coexistent hepatocellular carcinoma and cavernous hemangioma were correctly diagnosed by the following characteristic findings on dynamic CT and/or MR images; spreading and markedly prolonged enhancement in cavernous hemangioma, and the presence of capsule and/or transient diffuse enhancement in hepatocellular carcinoma. The third case of hepatocellular carcinoma and cholangiocarcinoma showed typical findings of hepatocellular carcinoma in one tumor and unusual complex enhancement in the other. CT and MR imaging may enable correct diagnosis to be made even in patients with multiple liver tumors of different kinds. PMID- 2555847 TI - Nervous system manifestations of Lyme disease. AB - Neurologic involvement is commonplace in Lyme borreliosis. Neuropathies can be acute or chronic, focal or disseminated, but are predominantly axonal. CNS infection can also be acute or indolent, focal or disseminated; meningitis, encephalitis, and cranial nerve palsies occur. A mild encephalopathy is also common, but only occasionally due to CNS infection. PMID- 2555848 TI - The pathogenesis of Lyme disease. AB - Lyme disease affects several major organ systems and leads to chronic illness. The pathogenesis of this disease appears to be centered around the long-term persistence of the organisms in tissues. In Lyme disease, isolations of B. burgdorferi are rare. It is thought that few organisms actually invade the host and that host mediators amplify the inflammatory response. Immune and nonimmune phagocytosis leading to bacterial killing occurs in Lyme disease. This organism shows preference for cell surfaces and tissues which may explain the paucity of isolations but also displays characteristic nonspecificity in its adherence to eukaryotic cells. This lack of specificity may explain its capacity to reside and injure vastly different tissues. Autoimmune mechanisms may coincide with spirochetal persistence in the pathogenesis of chronic Lyme disease. PMID- 2555849 TI - Treatment of Lyme borreliosis. AB - The infectious process of Lyme disease can appear as chronic dermatologic, rheumatologic, or neurologic. To rationally select a treatment regimen, the physician must have an appreciation of the clinical manifestations of the illness and of the systemic nature of the infection. The authors discuss the proper treatment protocols for each stage in the progression of Lyme disease. PMID- 2555850 TI - Lyme borreliosis in Europe. Neurologic disorders. AB - In Europe the tick-transmitted neurologic disorders MPN-GBB or Bannwarth's syndrome and ACA-associated neuropathy have been identified as clinical entities long before their causative agent was discovered. When Lyme disease and its neurologic manifestations were recognized in the United States, differences in the clinical pattern between North American and European cases with Lyme borreliosis were described in the initial reports. In the same way with the availability of serodiagnostic tests as the clinical spectrum of Lyme borreliosis was enlarging in Europe and in North America, these clinical differences became less prominent. PMID- 2555851 TI - Comparative effects of captopril and enalapril on the progression of chronic renal disease in partially nephrectomized rats. AB - Comparative effects of the angiotensin converting enzyme inhibitors captopril and enalapril on progression of chronic renal disease was studied in 3/4 nephrectomized rats. Rats were divided into sham and nephrectomized groups, and treated with plain water or water containing captopril (150 mg/liter) or enalapril (50 mg/liter). Evaluations were made 4 weeks before and 0, 4, 8, and 10 weeks after nephrectomy. Endogenous creatinine clearance decreased in drug treated, nephrectomized rats to values less than sham controls, but remained greater than water-treated rats. Significant (P less than 0.05) proteinuria developed 4 weeks post-nephrectomy in water-treated rats, 8 weeks post nephrectomy in captopril-treated rats, but did not develop in enalapril treated rats. Regression analysis of carbamylated plasma protein values vs plasma creatinine revealed significant (P less than 0.05) relationships only in the water-treated, nephrectomized rats from weeks 0 through 8, but were otherwise unaffected by treatment. Both drugs resulted in significantly (P less than 0.05) improved scores for renal histologic lesions as compared to water treatment. Modifications of proteinuria in captopril and enalapril-treated rats occurred prior to onset of changes in systolic blood pressure, which was significantly elevated only in water-treated, nephrectomized rats at weeks 8 and 10. We conclude that angiotensin converting enzyme inhibitors may ameliorate progression of experimental renal disease through intrarenal effects, independent of modulation of systemic blood pressure, and that enalapril may be superior to captopril in some regards. PMID- 2555852 TI - Intraduodenal administration of ethiofos (WR-2721): dose proportionality study in the rhesus monkey. AB - A dose progression crossover study of ethiofos (WR-2721) was conducted in three healthy male rhesus monkeys. Each subject was tested with three single intraduodenal doses containing 150, 300, and 600 mg/kg. Blood samples were drawn as a function of time and the concentrations of ethiofos, WR-1065 (free thiol metabolite), and total drug convertible to the free thiol (total WR-1065) were determined by HPLC using electrochemical detection. Ethiofos levels in plasma were usually below quantifiable limits of detection (0.23 mumol/L) at all three dose levels, but free WR-1065 plasma levels increased with increasing dose. Analysis of the free WR-1065 bioavailability values indicated large variability and an unpredictable dose response among subjects. Bound WR-1065 appears to reach saturable levels over the dose range, suggesting a saturable pool of binding sites in plasma. The time-to-peak plasma levels for WR-1065 were variable regardless of the administered dose and ranged from 1.0-2.5 hours. The high variability in the data may be a result of poor permeability or absorption of the parent compound (ethiofos), saturable binding to a variable pool of binding sites in plasma and/or high first-pass metabolism of ethiofos involving the gut lumen, gut wall (epithelium), and liver. PMID- 2555853 TI - Interactions of thyroid hormones with L-(3H) glutamate binding sites, with special reference to N-methyl-D-aspartate receptors. AB - An L-(3H)glutamate binding assay was developed in which 82% of the specific binding is to a site that corresponds to the N-methyl-D-aspartate receptor. The effects of various anticonvulsants and thyroid hormones were investigated with this assay. Anticonvulsants did not affect L-(3H)glutamate binding, while L- and D-isomers of triiodothyronine and thyroxine inhibited L-(3H)glutamate binding significantly. Scatchard analysis showed that thyroid hormones interacted with L (3H)glutamate binding competitively. The interactions of thyroid hormones with NMDA-sensitive L-(3H)glutamate binding may be one of the mechanisms by which these hormones affect the central nervous system. PMID- 2555854 TI - Effects of the anti-cancer agent etoposide on human herpesvirus 1 replication in vitro. AB - The anti-human herpesvirus type 1 (herpes simplex virus 1; HSV1) activity of etoposide (VP-16-213, a semi-synthetic derivative of epipodophyllotoxin) was investigated in vitro. Etoposide (but not the proprietary solvent in which the compound is usually formulated) demonstrated a significant antiviral action, probably through an effect on virus replication. Etoposide, at 3 micrograms/ml, induced a 50% reduction of HSV1-plaque formation in Vero cells. These findings are considered in the context of the use of etoposide in an in vivo procedure for the diagnosis of herpes encephalitis through virus-specific scintigraphic brain imaging. PMID- 2555855 TI - Molecular analysis of receptor binding and infectivity of the human immunodeficiency virus. PMID- 2555856 TI - Hemorrhagic shock: new vistas. Proceedings of an international symposium. November 18-19, 1988, Modena. PMID- 2555857 TI - Cholecystokinin peptides and bombesin reverse hemorrhagic shock in rats. PMID- 2555858 TI - Effect of ACTH-(1-24) on the volume of circulating blood and on regional blood flow in rats bled to hypovolemic shock. PMID- 2555859 TI - Characteristics of brain, heart ventricle and spleen capsule adrenoceptors in rats bled to hypovolemic shock and treated with ACTH-(1-24). PMID- 2555860 TI - Resuscitation fluid temperature: effect of delivery method. PMID- 2555862 TI - ACTH-(1-24) and hemorrhagic shock: preliminary clinical results. PMID- 2555861 TI - Beta-endorphin levels after experimental blood loss in human subjects. Correlations with cortisol, ACTH, plasma renin activity, plasma catecholamines and blood pressure variations. PMID- 2555863 TI - Preliminary data on the use of ACTH-(1-24) in human shock conditions. PMID- 2555864 TI - Disseminated intravascular coagulation and trauma. PMID- 2555866 TI - Hemorrhagic shock and anaesthesia: operative experience. PMID- 2555865 TI - Blood transfusion in hemorrhagic shock. PMID- 2555867 TI - Shock and traumatic retroperitoneal hematoma: diagnostic and therapeutic problems. PMID- 2555868 TI - On a case of shock caused by acute puerperal inversion of the uterus. PMID- 2555869 TI - Left extrapleural hemothorax from rupture of the subclavian artery. PMID- 2555870 TI - Endothelium-dependent responses and their potential relevance to the distribution of blood flow during hemorrhagic shock. AB - The endothelium releases vasoactive substances in response to several circulating hormones and to changes in shear stress of the flowing blood across the endothelial surface. Because the distribution of receptors mediating the release of endothelium-derived factors is not uniform along the vascular bed, the vascular endothelium has a potential role in regulating regional blood flow and maintaining peripheral resistance during the course and recovery from hemorrhagic shock. PMID- 2555871 TI - Beta-endorphin and central control of arterial blood pressure during challenge of circulatory homeostasis. AB - A variety of neurotransmitters and neuropeptides appear to participate in the central control mechanisms of arterial blood pressure. Our knowledge of these mechanisms is limited as yet. In the present study the involvement of the opioid peptide beta-endorphin in circulatory homeostasis was studied. Under conditions in which beta-endorphin does not affect basal blood pressure and heart rate this peptide had a pronounced prohypotensive influence in normotensive rats. This was found for two conditions during which circulatory homeostasis was challenged. Firstly, during blood letting in a rat model employed to test blood pressure regulation during hemorrhage, and secondly, for the central hypotensive action of alpha-methyldopa. In the first model hypotension was produced by stepwise bleeding to respectively 80, 60 and 40 mmHg mean arterial pressure. Intracerebroventricular (i.c.v.) administration of an antiserum raised against beta-endorphin or of naloxone (s.c. or i.c.v.) caused a significant increase in the required bleeding volume, whereas an opposite action was observed after the injection of morphine (s.c.) or of beta-endorphin (i.c.v.). The role of beta endorphin in the hypotensive action of alpha-methyldopa, given intracisternally (i.c.) was evaluated in conscious rats equipped with chronic cannulas. Pretreatment with the opiate antagonist naltrexone (i.c.) caused an inhibition of the hypotension and bradycardia induced by alpha-methyldopa. This effect of the receptor antagonist was mimicked by i.c. administration of a beta-endorphin antiserum. Taken together, these data point to a hypotensive influence exerted by endogenous beta-endorphin under conditions during which circulatory homeostasis are challenged. PMID- 2555872 TI - Neural influences on the human pulmonary circulation as a defence reaction to volume depletion. AB - The role of the autonomic nervous system in the regulation of pulmonary vasomotility in man is unsettled and great emphasis is usually given to changes in flow as the main regulating mechanism. In order to simulate hypovolemia, which might reduce the mechanical influence of flow and disclose a neural mechanism, we decreased venous return through balloon distention in the inferior vena cava in 12 normal subjects, during right heart catheterization performed for diagnostic purposes. Caval obstruction was graduated to reduce cardiac output, right atrial and pulmonary arterial pressures, without altering systemic arterial pressure and heart rate. The sympathetic nervous system was activated by arithmetic and cold pressor tests. During the former, the increase in cardiac output was more than halved by venous return restraint, as compared to the unrestrained condition, and clear pulmonary vasoconstriction, instead of vasodilatation, was observed. During the cold test, cardiac output remained almost steady, in the absence as in the presence of balloon expansion. In both conditions pulmonary arteriolar resistance rose, but in the latter this increase was more than doubled. This study suggests that the autonomic nervous system is involved in the regulation of pulmonary vasomotility in man, its role being unveiled when the mechanical influence of flow is reduced by mimicking a hypovolemic state. PMID- 2555873 TI - Oxygen-derived free radicals in the pathogenesis of experimental shock. AB - Oxygen-derived free radicals are naturally produced in biological systems mostly in ischemia and hypoxia related conditions. Imbalance of physiological defenses against oxygen-derived free radicals causes cellular damage. In our laboratory the role of oxygen radicals in the pathogenesis of circulatory shock was studied by exploiting the scavenging action of the spin-trapping compound phenyl-butyl nitrone (PBN) in experiments concerning: (1) survival after shock, (2) microcirculatory derangements in endotoxin shock, (3) fluidity modifications of cell membranes during shock, (4) exhalation of ethane as non-invasive marker of shock. In some experiments the steroid methylprednisolone was used. Results showed that administration of PBN and of the steroid (1) ensures survival after otherwise lethal shock as confirmed by decreased ethane exhalation, (2) prevents microcirculatory troubles, (3) maintains stability of cell membranes. These findings strongly support the role of oxygen-derived free radicals in the pathogenesis and pathophysiology of circulatory shock. PMID- 2555874 TI - Platelet-activating factor and cytokine interactions in shock. AB - Many types of shock are characterized by profound hemodynamic alterations and depression of immune processes. Among the various mediators implicated in shock conditions, there is much evidence to suggest that, together with various cytokines, the inflammatory and chemotactic autacoid, platelet-activating factor (PAF), plays an important role. Studies on several animal models have shown that infusion of PAF mimicks the shock state, that markedly increased levels of PAF are produced in shock and that PAF antagonists afford significant protection against diverse forms of shock. The precise mechanism by which PAF antagonists protect against shock remains unclear; however, it is becoming apparent that in traumatic states a complex interaction occurs between PAF and cytokines, which leads to the acute phase reaction and circulatory collapse. We propose that PAF antagonists may be effective in counteracting shock because of their antiprotease activity and their ability to inhibit deleterious PAF/cytokine auto-generated feedback processes. PMID- 2555875 TI - Sympathetic and hemodynamic adjustments to hemorrhage: a possible role for endogenous opioid peptides. AB - The traditional view of the decrease in blood pressure during blood loss is that it is a passive phenomenon. Blood pressure falls due to the inability of the compensatory mechanisms to keep pace with blood loss. However, recent evidence indicates that this transition from normotension to hypotension may involve an active decrease in compensation by the sympathetic nervous system. In the conscious, chronically prepared rabbit, blood pressure is maintained early in hemorrhage primarily by sympathetically mediated compensatory increases in vascular resistance and heart rate. When blood loss exceeds approximately 20% of total blood volume, hypotension develops abruptly due to a decrease in vascular resistance. This vasodilation is accompanied by decreased plasma norepinephrine (NE) levels and decreased sympathetic nerve activity. Therefore, the transition from normotension to hypotension during hemorrhage involves an active change, a decrease in vascular resistance, associated with a decrease in sympathetic nerve activity. Opioid receptor blockade with naloxone reverses acute hemorrhagic hypotension by increasing vascular resistance. The increase in resistance is accompanied by increased plasma NE and increased sympathetic nerve activity. Thus, the transition to hypotension during acute hemorrhage may be an active event brought on by a decrease in sympathetic outflow. Naloxone's reversal of the hypotension is consistent with the central involvement of endogenous opioid peptides in this phenomenon and thus in the pathogenesis of hypotension during blood loss. PMID- 2555876 TI - Facilitation of the release of noradrenaline: an extra-adrenal effect of adrenocorticotropic hormone. AB - Effects of ACTH-(1-24) on the release of noradrenaline from postganglionic sympathetic neurones were studied in rabbits. In the isolated perfused rabbit heart, ACTH-(1-24) (0.1-100 nmol/l) increased the overflow of noradrenaline elicited by sympathetic nerve stimulation at 1 Hz, an effect that persisted in the presence of propranolol and cocaine. In pithed rabbits with electrically stimulated sympathetic outflow (2 Hz), ACTH-(1-24) (0.03-1 microgram/kg per min) increased the plasma noradrenaline concentration as well as the calculated rate of entry of noradrenaline into the plasma, but decreased arterial blood pressure by a direct vasodilator effect. In anaesthetized rabbits, ACTH-(1-24) (0.1-1 microgram/kg per min) also increased the plasma noradrenaline concentration; the higher dose in addition reduced the blood pressure and increased the firing rate of renal sympathetic nerves; however, the rise in plasma noradrenaline was greater than would have been expected from the increase in sympathetic firing rate alone. It is concluded that ACTH exerts two primary effects on the cardiovascular system of the rabbit: presynaptic facilitation of the noradrenaline release and postsynaptic vasodilation. The vasodilation leads to a baroreceptor-mediated reflex increase in sympathetic tone. The low concentrations of ACTH required suggest that presynaptic facilitation may occur in vivo, at least when the secretion of ACTH is high. PMID- 2555877 TI - The therapeutic efficacy of opiate antagonists in hemorrhagic shock. AB - The endogenous opioids have been implicated as contributing factors to the cardiovascular dysfunction of shock. Opiate receptor antagonists improve cardiovascular function and long-term survival in laboratory animal models of shock. In this communication, evidence of the therapeutic efficacy of opiate antagonists in canine and primate hemorrhagic shock is presented. The animals were hemorrhaged into a reservoir to lower MAP to 45 mmHg and that pressure was maintained for 1 h at which time the reservoir was clamped and treatment initiated. The "shed blood" was returned at t = 120 min and treatment continued until t = 180 min. Opiate antagonists employed included naloxone, naltrexone and the mixed agonist/antagonist agent, nalbuphine. Both naloxone and naltrexone improved cardiac function at doses of 1 and 2 mg/kg. Animal survival was significantly enhanced in the high dose format. Nalbuphine also improved cardiovascular performance at doses from 1 to 4 mg/kg but at higher doses it depressed cardiac performance. The efficacy of the antagonists is attenuated by acidosis and hypothermia. Opiate antagonists may induce cardiac arrhythmias in combination with beta-adrenergic blocking drugs and the efficacy is reduced in animals that received high dose steroid therapy. Thus the use of opiate antagonists would be contraindicated in patients that received drugs such as propranolol or methylprednisolone. There have been no controlled clinical trials of opiate antagonists in human hemorrhagic shock; these are needed for final clarification. PMID- 2555878 TI - The adrenocorticotropic hormone (ACTH)-induced reversal of hemorrhagic shock. AB - Adrenocorticotropic hormone (ACTH), while having negligible effects on cardiovascular function in the intact animal, induces a potent and sustained reversal of an otherwise invariably, rapidly fatal condition of hemorrhage induced hypovolemic shock, in rats and dogs. The main site(s) of action are at the peripheral level; however, subsidiary site(s) of action in the CNS cannot be excluded. The studies on the mechanism of action indicate that the ACTH-induced reversal of hemorrhagic shock (a) is an extra-hormonal, adrenal-independent effect, because it is not affected by adrenalectomy and is shared by many ACTH fragments practically devoid of corticotropic activity; (b) is antagonized by morphine in a surmontable way; (c) needs the functional integrity of the sympathetic nervous system (it is prevented by guanethidine, reserpine, and clonidine) and the availability of peripheral alpha-adrenoceptors (it is antagonized by dibenamine, prazosin and yohimbine, but not by practolol); (d) requires the integrity of afferent vagal fibers (it is almost completely abolished by vagotomy); (e) involves central cholinergic networks (it is antagonized by atropine sulphate, but not by atropine methyl bromide; and it is prevented by the intracerebroventricular injection of hemicholinium-3); (f) is associated with a massive increase in the volume of circulating blood, likely due to a mobilization from peripheral pooling sites (it is largely prevented by splenectomy or by suprahepatic veins ligature, and is associated with a restoration of the venous blood flow in peripheral vascular beds and with a normalization of venous PO2); (g) is associated with a restoration of heart and spleen adrenoceptors, whose number is significantly decreased during hemorrhagic shock. The survival time of hemorrhage-shocked animals, which is 26 +/- 3 min in controls, is greatly prolonged (44 +/- 18 h) by ACTH, provided that the treatment is made within 5-10 min after bleeding. Finally, in animals treated with ACTH within 5-10 min after bleeding, blood reinfusion retains its effectiveness and reverse shock even if performed 2-5 h later. PMID- 2555879 TI - The C1 area of rostral ventrolateral medulla: a central site integrating autonomic responses to hemorrhage. AB - Activation of the sympathetic neurons and release of adrenomedullary catecholamines are the principal early reflex responses to hemorrhage. These are initiated by arterial baro- and chemoreceptors, from other cardiopulmonary receptors, and by intracerebral receptors responding to ischemia. A principal gateway for integrating the autonomic responses are a small collection of neurons in a region of the rostral ventrolateral medulla (RVL), containing a cluster of neurons of the C1 adrenergic cell group, the C1 area. Neurons in the C1 area of RVL project exclusively to autonomic nuclei of the spinal cord, are tonically active, and fire with a rhythm linked to the cardiac cycle. They are essential for maintaining resting discharge of sympathetic nerves and, consequently, arterial pressure (AP) and heart rate. They also are critical for reflex changes in AP in the baro- and chemoreceptor, somato-sympathetic (pain), and cerebral ischemic reflexes. Neurons of the C1 area are under tonic excitatory and inhibitory control by pathways from other autonomic centers. They are controlled by a range of neurotransmitters including, gamma aminobutyric acid (GABA), acetylcholine, catecholamines, enkephalin, and several neuropeptides. They also serve as a site of action for the hypotensive actions of several clinically important neurotransmitters. The C1-area of RVL may play a critical role in the autonomic responses to hemorrhage and may be an important target for drugs seeking to treat hemorrhagic shock. PMID- 2555880 TI - Adrenocorticotropic hormone (ACTH) and centrally-acting cholinomimetic drugs improve survival of rats with severe hemorrhagic shock through distinct central cholinergic mechanisms. AB - Pharmacological doses (40-160 micrograms/kg) of adrenocorticotropic hormone (ACTH) intravenously injected to urethane-anesthetized rats subjected to otherwise lethal hemorrhagic shock (mean arterial pressure stabilized at 20-25 mmHg) promptly restore blood pressure to about the pre-bleeding values, and prevent death (anti-shock effect). Hemicholinium-3 (i.c.v. injected) and atropine sulphate, but not atropine methylbromide, antagonize these ACTH effects. Moreover, since pirenzepine, injected i.v. or i.c.v., does not affect the anti shock activity of ACTH, the central cholinergic mechanism participating in this ACTH action must involve M2, but not M1 brain muscarinic receptors. Intravenous physostigmine, too (but not neostigmine) and oxotremorine have an ACTH-like anti shock effect, which however is neither affected by hemicholinium-3, nor by atropine methylbromide, nor by atropine sulphate, but only by high i.c.v. doses of gallamine or pancuronium. On the other hand, reserpine, guanethidine, and alpha-adrenoceptor blocking drugs inhibit the anti-shock effect of ACTH as well as that of oxotremorine and physostigmine. It is suggested that, in rats, both ACTH and cholinergic drugs must activate a central cholinergic mechanism(s) in order to exert a sympathetic nerve-mediated anti-shock effect. However, receptors involved are of the muscarinic M2 subtype in the case of ACTH, and probably nicotinic in the case of cholinergic drugs. That ACTH and cholinergic drugs activate different central cholinergic mechanisms is also suggested by the fact that cholinergic drugs have a centrally-mediated hypertensive action in normal animals, which is not shared by ACTH. PMID- 2555881 TI - Effects of fructose 1,6-diphosphate on splanchnic artery occlusion shock in the rat. AB - Splanchnic artery occlusion (SAO) shock, produced by clamping splanchnic arteries for 45 min followed by the release of occlusion, was induced in male rats, treated 15 min before surgery, with fructose 1,6-diphosphate (FDP) or with equivalent doses of fructose or inorganic phosphate. Survival rate, peritoneal macrophage phagocytosis and plasma levels of myocardial depressant factor (MDF) were measured. Shocked animals pretreated with vehicle exhibited 24.6 +/- 0.9% phagocytic activity, 110 +/- 3.9 units/ml MDF plasma levels and 0% survival. Sham animals showed the following values: survival 100%; phagocytosis, 49.5 +/- 1.3%; MDF, 22 +/- 2.9 units/ml. Pretreatment with FDP (25 mg/kg/i.v.) significantly improved survival rate (50%) and macrophage phagocytosis (37.9 +/- 0.4%) and reduced plasma MDF levels (77 +/- 3 units/ml). Equivalent doses of fructose and inorganic phosphate did not improve survival, as well as lower doses of FDP. These results suggest a beneficial effect of FDP in SAO shock. PMID- 2555882 TI - Angiotensin II and atrial natriuretic peptide in the brain: effects on volume and Na+ balance. AB - Atrial natriuretic peptide (ANP) and angiotensin II (ANG II), although originally isolated from peripheral sources, are now known to be present in the central nervous system. The distribution of the peptides and their binding sites are found in areas involved in cardiovascular and volume/electrolyte regulation. Since ANP administered centrally can antagonize the actions of ANG II, the two peptides may function as opposing mechanisms involved in maintaining fluid and electrolyte homeostasis. PMID- 2555883 TI - New prospects for shock treatment. PMID- 2555884 TI - Brain resuscitation. AB - Optimal neurological outcome after cardiac arrest requires careful attention to the details of both intracranial and extracranial homeostasis. A high index of suspicion regarding the potential causes and complications of cardiac arrest facilitates discovery and treatment of problems before they adversely impact upon neurological outcome. The future is bright for resuscitation research since our fundamental understanding of cerebral ischemia and its consequences has dramatically improved. This knowledge can hopefully be transferred to clinical useful modes of therapy. PMID- 2555885 TI - Advances in prehospital immediate care. AB - Although studies documenting the influence of prehospital care on patient outcome are most urgently needed, recent techniques and innovations being examined in the field setting may improve the clinical care delivered. The basic manoeuvres for airway and breathing have been in need of emphasis for some time. Sellick's manoeuvre has been resuscitated and is being emphasized in most training programmes. Bag-mask ventilation, notoriously difficult in field settings in the hands of emergency medical personnel, has been improved with the recent improvement of mask design. Endotracheal intubation has been shown to influence patient outcome, and can be performed with surprising success by paramedics. Complication rates continue to cause concern, and a study to document the incidence of tube misplacement has just begun in the United States and Canada. Transillumination and translaryngeal jet ventilation have been refined as adjunct techniques to field intubation. Patient monitoring has improved through the more common use of pulse oximetry and a recently-introduced "CO2 Detector" that is designed to indicate via a colour change, oesophageal placement of an endotracheal tube. Automatic external defibrillators have come under some scrutiny, and early data suggest their safe use by lay persons, a minimal effect on outcome in high-risk patients, and a significant effect on outcome only when combined with other essential elements of prehospital care. The role of IV fluids in prehospital trauma care has been clarified, as well as that of the pneumatic antishock garment. Transcutaneous pacemakers have been developed and shown effective if used early, and thrombolytic therapy has been examined for the prehospital management of acute myocardial infarction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555886 TI - Monitoring in emergency medicine. AB - Clinical observation is the most valuable monitoring technique we have. Complexity and invasiveness of monitoring increases from prehospital care to Emergency Department, to Anaesthesia and Intensive Care. Many methods of monitoring have specific applications. Non-invasive blood pressure monitoring has no advantages over conventional cuff methods, other than freeing the hands of the operator. Non-invasive cardiac output measurement, transcutaneous oxygen and carbon dioxide measurement are unlikely to play a major role in the foreseeable future in the emergency setting. The most exciting development in recent years has been the widespread availability of pulse oximetry, which allows beat by beat analysis of haemoglobin oxygen saturation. PMID- 2555887 TI - Surfactant therapy. AB - This paper presents the rationale for surfactant replacement therapy. In 6 animal models in which respiratory failure was induced by damaging the bronchial surfactant, surfactant depletion by lung lavage, by free oxygen radicals, by influenza virus and by anti-lung serum, it is demonstrated that surfactant instillation always led to an improvement in gas exchange and lung mechanics. A first clinical trials shows that surfactant instillation also improved lung function in a patient with severe respiratory failure. It is speculated that surfactant replacement therapy will become an acceptable therapeutic measure for treating lung failure in patients suffering with ARDS. PMID- 2555888 TI - Microcirculatory therapy in shock. AB - The normal microvascular perfusion pattern is characterized by temporal and spatial variations of capillary flow. Local driving pressure, arteriolar vasomotion and endothelial cells are key-factors for local regulation of hydraulic resistance and fluid balance between the blood and tissue compartments. In shock, both the central and particularly the local mechanisms controlling microvascular perfusion are impaired. The microvascular perfusion pattern becomes permanently inhomogeneous due to lack of arteriolar vasomotion, changes of flow properties of blood, endothelial cell swelling and blood cell-endothelium interaction. Hence the objectives of primary shock therapy are to reestablish precapillary pressure, arteriolar vasomotion and to open the occluded microvascular pathways in order to reestablish the surface area needed for exchange of nutrients and drainage of waste product. These effects can not be achieved by vasoactive drugs, unless blood volume has been restored and blood fluidity improved by hemodilution. Whereas the necessary hemodilution can be achieved by conventional volume substitutes (colloids, crystalloids) restoration of vasomotion and reopening of narrowed capillaries can be obtained by small volume resuscitation using hyperosmotic/hyperoncotic salt dextran solution. The potential of this new concept for primary resuscitation and treatment of tissue ischemia is presently explored. PMID- 2555889 TI - Role of oxygen radicals and antioxidants in adult respiratory distress syndrome. Potentials in therapy. PMID- 2555890 TI - Cell therapy in shock: a reasonable prospect or a lost cause? AB - Almost any insult to the body can set in motion the sequence of physiologic and biochemical changes leading to shock. In the absence of effective treatment widespread disruption of the body's biological mechanisms inevitably follows. This review will describe some of cellular and subcellular disturbances associated with the shock syndrome, the persistence of which lead to (multiple) organ failure. Current views as to the mechanisms of these disturbances will be outlined with particular reference to hypovolemic and septic shock. Finally, an indication will be given of the relevant therapeutic implications, now and in the future. PMID- 2555891 TI - [The herbalist pastor Johann Kunzle and the medical community]. AB - The famous herbalist, Johann Kunzle, was not only a catholic priest but also a popular healer. A quack doctor with a great deal of psychological skill, he published numerous books and journals on the use of medicinal herbs. Over one million copies of his bestselling book "Chrut and Uchrut" were sold in German alone. With his outstanding knowledge of herbs he literally created an industry out of herbal remedies. Opinions about the legendary "herb Father" differed considerably and still do today. Some consider that he was a great philanthropist, others that he was merely a charlatan and clever businessman. Certainly his activities as one of the most famous exponents of alternative medicine in Swiss history were regarded with deep suspicion by the medical profession. The success of Johann Kunzle and his remedies emphasises the importance of psychological factors both in sickness and in health and also the irrational belief which so many people tend to have about illness and health. PMID- 2555892 TI - [Epidemiology of human papillomavirus genital infections in women]. AB - Epidemiological variations condylomata plana show annual fluctuations and vast geographical differences. The risk is obviously related to the number of sexual partners and their profession. PMID- 2555893 TI - [Diagnosis of papillomavirus infection of the female genital system]. AB - Cytology and colposcopy are currently the only diagnostic means of papilloma virus lesions. The virologic diagnosis is still at the research stage. The treatment of the lesion usually consists in destruction by physical means. PMID- 2555894 TI - [Use of Polygynax in pregnant women for preventing cervico-vaginal infections]. AB - 115 women, distributed in 20 centres, were included in a randomized study designed to assess the efficacy of Polygynax in preventing vaginal infections in women at risk, at the start of pregnancy. Polygynax proved to be markedly effective since only 9.6% of the patients in the Polygynax group had a vaginal infection, in contrast to 42.6% of the patients in the control group. This result is highly significant (p less than 0.0001) and confirms the advantage attached to the use of Polygynax when a first line treatment involving the use of several drugs in combination is indicated. PMID- 2555895 TI - [Almitrine (Vectarion)--an always up-to-date respiratory adjuvant]. PMID- 2555896 TI - [Current aspects of the antitubercular chemotherapy of new patients with tuberculosis and diabetes mellitus]. AB - Starting from the published data that demonstrate the annulling of the negative influence of diabetes-tuberculosis morbid association, the authors studied from the clinical, epidemiologic, paraclinical and therapeutical viewpoints 68 diabetic patients with pulmonary tuberculosis (31 men, 37 women) and a control group of 73 patients (57 men and 16 women), both groups were new cases under a first treatment. There was no difference as regards the onset, evolution up to the moment of detection and diagnosis, the presenting clinical and radiologic forms, the gravity of the cases, biologic test and evolution under treatment. These data demonstrate the efficiency of the present day treatment, regardless of the association of the two diseases, the annulling of the negative influence described in previous years, but also the lability of diabetic patients in spite of the control of tuberculosis by tuberculostatics. PMID- 2555897 TI - [The relation of severe obstructive disorders to ventilation found in young patients with bronchitis and bronchopulmonary diseases in childhood]. AB - Of the patients with chronic obstructive pulmonary disease (COLD) and severe obstructive syndrome, 39 whose age was under 40 were selected. In 23 of them, the anamnesis revealed bronchopulmonary affections in childhood, that required admission into the hospital (19 were non-smokers). Of the rest of 16 patients, 14 were hard smokers that started to smoke before the age of 14. The functional picture was severely modified, similarly to that of the COLD patients in the 6th decade of life. This suggests that the degradation process started in the childhood, and that the chronic respiratory diseases and/or smoking at an early age had an important role. PMID- 2555898 TI - [The current situation of mycobacterial chemoresistance and its implications for antitubercular treatment]. AB - The data in the present paper consider, for a longer period, the secondary and the initial drug resistance in pulmonary and extrapulmonary tuberculosis. The paper studies mainly the period of the last 9 years (1979-1987) in comparison with a previous period of 6 years (1973-1978). The 2,194 antibiograms performed for 7 common tuberculostatics show an increase, in the last years, of the general resistance of the germs from 24.3% to 36%. The increase of the resistance is noticed to all the tuberculostatics, but mainly to Rifampicin, from 14.8% to 49.3% and to streptomycin, from 22.7% to 44.8%. An important increase is found, at present, mainly in the drug resistance of germs in the extrapulmonary specimens (lymph nodes 28.5% and CSF 40%) to streptomycin and INH. PMID- 2555900 TI - [Conclusions drawn about the current situation of antitubercular chemotherapy in Romania based on the 1987 national evaluation questionnaire]. PMID- 2555899 TI - [Coma--accidental poisoning occurring in the isoniazid + rifampicin treatment of a 7-year-old child]. PMID- 2555901 TI - [Short-term treatment (3+3) in pulmonary tuberculosis in adults]. PMID- 2555902 TI - [The late results of intermittent chemotherapy in primary tuberculosis in children (1969-1988)]. PMID- 2555903 TI - [An evaluation of the results of the use of a short-duration chemotherapy regimen (3+3) in cases with tuberculosis of the respiratory tract in 1986]. PMID- 2555904 TI - [Great pioneers in phthisiology. The centenary of the Carlo Forlanini (1847-1918) method]. PMID- 2555905 TI - [The effect of short-term treatment on the evolution toward chronicity of pulmonary tuberculosis]. PMID- 2555906 TI - [Results obtained with the first group of tuberculosis patients included in short duration intensive treatment]. PMID- 2555907 TI - [Patient noncompliance under conditions of short-term treatment (3+3) in the area of the Bacau TB Dispensary]. PMID- 2555908 TI - [Compliance with antitubercular chemotherapy, a central element in the efficacy of a national program]. PMID- 2555909 TI - [New tuberculostatics under experiment. Results and prospects]. PMID- 2555910 TI - Clostridium perfringens septicemia with massive hemolysis. AB - Massive hemolysis with acute renal failure occurred in a previously healthy 69 year-old patient as a complication of Clostridium perfringens septicemia secondary to gall bladder empyema. To our knowledge, this is one of the few patients with C. perfringens septicemia and massive intravascular hemolysis who survived the episode and regained a normal renal function. PMID- 2555911 TI - Norwalk virus in Norway: an outbreak of gastroenteritis studied by electron microscopy and radioimmunoassay. AB - An outbreak of acute gastroenteritis affected approximately half of 40 children staying at a holiday centre in Southern Norway. By direct electron microscopy Norwalk-like viruses were demonstrated in 4/8 available stool specimens. No other pathogens were detected. Antibody against these viruses was demonstrated by immune electron microscopy in all of 7 convalescent phase sera but in none of 11 acute phase sera collected. Radioimmunoassay examination showed a rise in titre of Norwalk virus antibody in 6 available paired sera. This outbreak of Norwalk virus gastroenteritis in Norway was thus documented by a combined use of direct and immune electron microscopy and radioimmunoassay. PMID- 2555912 TI - Coincidence of deficient delayed hypersensitivity and intestinal protozoa in homosexual men. AB - Stool samples of 134 subjectively and apparently healthy male homosexual volunteers were studied, and in 86 men (64.2%) 190 parasite infestations, up to 6 per host, were identified. The most frequent species were Endolimax nana (40), Entamoeba coli (38), Entamoeba histolytica (34), Entamoeba hartmanni (32), and Iodamoeba butschlii (22); Cryptosporidium was not found. All 11 isolates of Entamoeba histolytica analysed belonged to a noninvasive zymodeme: 10 were of zymodeme I and I of zymodeme III. In 172 healthy control adults 6 parasite infestations were found. In the homosexual men, a negative tuberculin test was more frequent in the presence than in the absence of parasites. Entamoeba histolytica (p = 0.005) Entamoeba coli (p = 0.013) and Iodamoeba butschlii (p = 0.004) were associated with the anergy. PMID- 2555913 TI - Disseminated histoplasmosis in a Danish patient with AIDS. AB - We present the first case of disseminated histoplasmosis in an AIDS patient in Europe, a 33-year-old Danish homosexual man, and recommend a detailed travel history in HIV-positive patients presenting with fever, weight loss and organomegaly. In Scandinavia disseminated histoplasmosis is rare but should be kept in mind as the disease is a major opportunistic infection in patients with AIDS. Treatment with amphotericin B followed by fluconazole was effective. PMID- 2555914 TI - [Factors affecting gap junction formation and permeability]. PMID- 2555916 TI - [Presynaptic receptors]. PMID- 2555915 TI - [Therapeutic effect of naloxone and its mechanisms in circulation shock]. PMID- 2555917 TI - The endocrinopathy of POEMS syndrome. AB - We describe the clinical features and post mortem findings in a patient with POEMS syndrome (P--polyneuropathy, O--organomegaly, E--endocrinopathy, M--M protein, S--skin changes), with particular emphasis on the evolution of the endocrinopathy over a three year period. In addition to gynaecomastia this case exhibited primary hypothyroidism and primary and secondary adrenal and gonadal failure. The widespread nature of the endocrine changes suggests that a circulating substance interferes with the action of trophic hormones, possibly by inhibiting second messenger production. PMID- 2555918 TI - Failure of somatostatin analogue SMS 201-995 to control Cushing's syndrome due to ectopic release of ACTH from a bronchial carcinoid. AB - A patient with ectopic Cushing's syndrome secondary to a malignant thymic carcinoid tumour was treated with the somatostatin analogue SMS 201-995. The administration of the drug subcutaneously in increasing doses over a 34 day period failed to control the ACTH or glucocorticoid excess. PMID- 2555919 TI - Gastric rupture due to excessive sodium bicarbonate ingestion. PMID- 2555921 TI - Sphingomyelin synthase and PKC activation. PMID- 2555920 TI - DNA topoisomerase I--targeted chemotherapy of human colon cancer in xenografts. AB - Drug development is needed to improve chemotherapy of patients with locally advanced or metastatic colon carcinoma, who otherwise have an unfavorable prognosis. DNA topoisomerase I, a nuclear enzyme important for solving topological problems arising during DNA replication and for other cellular functions, has been identified as a principal target of a plant alkaloid 20(S) camptothecin. Significantly increased concentrations of this enzyme, compared to that in normal colonic mucosa, were found in advanced stages of human colon adenocarcinoma and in xenografts of colon cancer carried by immunodeficient mice. Several synthetic analogs of camptothecin, selected by tests with the purified enzyme and tissue-culture screens, were evaluated in the xenograft model. Unlike other anticancer drugs tested, 20(RS)-9-amino-camptothecin (9-AC) induced disease free remissions. The overall drug toxicity was low and allowed for repeated courses of treatment. PMID- 2555922 TI - Progress in vaccines against AIDS. PMID- 2555924 TI - [Electrophysiologic comparison between sutured and glued nerve anastomoses]. AB - Fibrinadhesires are successfully used in many areas of surgery, but they have not been generally adopted for the construction of neural anastomoses, even though the conventional suturing technique is also not completely satisfactory. In the early phase of fibrinotherapy, dehiscence occurred. The addition of antifibrinolytics has admittedly led to stabilization of the clots, but since the introduction of this procedure fibrosis has been observed with increasing frequency. This can cause even initially successful anastomoses to fail later. The objective of the work described in the present paper was to compare the conventional suturing technique and fibrinotherapy for the fixation of anastomoses. Electrophysiological methods of measurement were used, as it is ultimately the degree of reinnervation achieved that is of decisive importance. No significant difference was noted when an epineural technique was used. PMID- 2555923 TI - A formalin-inactivated whole SIV vaccine confers protection in macaques. AB - A vaccine against human immunodeficiency virus (HIV) would be highly effective in stopping the acquired immunodeficiency syndrome (AIDS) epidemic. A comprehensive evaluation of potential vaccine methodologies can be made by means of the simian model for AIDS, which takes advantage of the similarities in viral composition and disease potential between simian immunodeficiency virus (SIV) infection of rhesus macaques and HIV infection in humans. Immunization with a formalin inactivated whole SIV vaccine potentiated with either alum and the Syntex adjuvant threonyl muramyl dipeptide (MDP) or MDP alone resulted in the protection of eight of nine rhesus monkeys challenged with ten animal-infectious doses of pathogenic virus. These results demonstrate that a whole virus vaccine is highly effective in inducing immune responses that can protect against lentivirus infection and AIDS-like disease. PMID- 2555925 TI - A study on the usage of respirators among granite quarry workers in Singapore. AB - The frequency and correctness of respirators were studied in 5 granite quarries in Singapore involving 201 workers. The overall prevalence of usage of correct respirators was 45.8%. 10.4% of the workers were found to be using the wrong respiratory protective devices. Age, years of exposure and types of occupations were found to affect the usage of respirators. Some common reasons given by workers for not wearing the respirators were 'breathing difficulty', 'hot & sweaty', and 'respirator smells after a while'. PMID- 2555926 TI - Natural history of atlanto-axial subluxation in rheumatoid arthritis. AB - In 1969 the authors started their prospective study to determine the natural history of atlanto-axial subluxation in patients suffering from rheumatoid arthritis. Between January 1969 and July 1971 they accumulated 41 cases of atlanto-axial subluxation. These patients were studied clinically and radiologically, and the initial findings were published in the Journal of Bone and Joint Surgery, Volume 55B, August 1973. All of these patients were followed clinically and radiologically until 1981 or until their demise. Roentgenographically, 61% have remained unchanged, 27% have shown progression of the atlanto-axial subluxation, and 12% showed decrease in the amount of atlanto axial subluxation, including one following surgery. During this 10-year period, 12 patients with atlanto-axial subluxation have died, two with historical evidence of neurologic damage. The remaining 10 patients have died from unrelated causes. Only 3 patients underwent surgical stabilization. This study concluded that the atlanto-axial subluxation is compatible with life, but some patients require surgical intervention. PMID- 2555927 TI - Bovine parathyroid tissue: a model to compare the biosynthesis and secretion of parathyroid hormone and parathyroid hormone-related peptide. AB - Bovine parathyroid tissue was evaluated as a model to compare parathyroid hormone related peptide (PTH-rP) and parathyroid hormone (PTH) secretion. Tissue was incubated in variable calcium levels (n = 5). A parathyroid cell digest was prepared from collagenase-treated glands. PTH-rP and PTH levels were determined by radioimmunoassay. PTH-rP bioactivity was determined by 3H-cAMP production in a UMR 106 cell bioassay. PTH-rP levels in the incubation medium were 2.0 ng/mg protein (0.25 mmol Ca++), 2.2 ng/mg protein (1.25 mmol Ca++), and 1.9 ng/mg protein (2.5 mmol/L Ca++). PTH levels were 321 ng/mg protein (0.25 mmol/L Ca++) and 200 ng/mg protein (2.5 mmol Ca++). Therefore, calcium significantly inhibited PTH but not PTH-rP secretion (p = 0.03). Addition of incubation medium to the bioassay resulted in 3H-cAMP levels that were 8 to 10 times greater than basal levels. Greater than 50% of the activity persisted after addition of PTH antibody, demonstrating that a significant amount of the activity was caused by PTH-rP. Tissue PTH-rP was 5.1 ng/mg protein, compared with 2080 ng/mg protein for PTH. We conclude that (1) bovine parathyroid tissue contains bioactive PTH-rP and is a useful model to compare the biosynthesis and secretion of PTH-rP and PTH in normal tissue and (2) unlike PTH, PTH-rP secretion is not regulated by calcium. PMID- 2555928 TI - Increased calcium levels alter cellular and molecular events in wound healing. AB - Surgical morbidity is dictated directly by wound healing. We have studied the effects of elevated calcium levels using cultured keratinocytes in vitro on two of the rate-limiting steps of wound healing, chemotaxis (directed migration) and adhesion. We found that the increased calcium (10 mmol/L) significantly inhibited both keratinocyte chemotaxis and adhesion (p less than 0.05). The calcium effect on adhesion could be partially reversed by pretreatment with the calcium channel blocker verapamil. Based on these data, an animal model was formulated in which topical calcium (5 mmol/L/day) was added to linear incision wounds. This resulted in significantly (p less than 0.05) delayed wound contraction characteristic of a chronic or impaired wound. Wound contraction depends on the presence of fibroblasts that synthesize collagen. The chronic wound was characterized by increased collagenase activity (p less than 0.05) but little alteration in collagen I synthesis. The addition of verapamil to these chronic wounds resulted in improved wound closure. These studies define the molecular and cellular events occurring as a result of the addition of elevated levels of calcium both in vitro and in vivo. Calcium may play a key role in the pathogenesis of chronic wounds. PMID- 2555929 TI - Tumor necrosis factor: immune endocrine interaction. AB - Tumor necrosis factor (TNF), a peptide produced by macrophages in response to endotoxin, has been implicated as a mediator of septic shock. This study examined the effects of injections of recombinant (r) human TNF on circulating levels of metabolic substrates and hormones in conscious, unrestrained rats and the effects of TNF on cortisol secretion from human adrenocortical cells in vitro. Sublethal doses of rTNF--doses that did not produce hemodynamic changes in previous work- produced rapid (1 hour), significant increases in blood levels of glucose, lactate, and triglycerides and decreases in plasma levels of branched chain amino acids. Plasma levels of glucagon, corticosterone, ACTH, norepinephrine, and dihydroxyphenylglycol were also increased significantly. Incubation of adrenocortical cells with either 0.15 or 1.5 micrograms of rTNF increased cortisol secretion to the same extent as did 10(-10) mol/L ACTH. Administration of TNF produces a variety of metabolic and neuroendocrine effects including stimulation of anterior pituitary, adrenal cortical, and pancreatic secretion, and sympathoneural activation. These changes, and the in vitro results, are consistent with the view that immune cells can interact with endocrine cells through release of TNF. PMID- 2555930 TI - [T1-2N0M0-stage lung cancer, its diagnosis and treatment results]. PMID- 2555931 TI - [The role of an endogenous digoxin-like factor in regulating blood circulation and in the origin of arrhythmia in myocardial ischemia]. AB - The history of the discovery of endogenous digoxin-like factor (EDF) is described and the role played by the substance in blood circulation regulation, in the pathogenesis of arterial hypertension is discussed. The authors provide their own data (both experimental and clinical ones) concerned with EDF participation in the pathogenesis of early ventricular fibrillations in acute myocardial ischemia. Experiments on rats demonstrated that myocardial infarction (MI) is marked by a negative linear correlation between the intensity of ventricular fibrillations and the activity of Na,K-ATPase of intact red blood cells (r = -0.84) that mirrors the content of circulating EDF. Administration to the animals of digoxin antibodies binding EDF resulted in the antiarrhythmic effect and in the recovery of the enzyme activity. The patients demonstrated, within the first day of MI, a 76-percent inhibition of the activity of Na,K-ATPase of red blood cells. A correlation was discovered between the enzyme activity and the capacity of protein-free supernatants of blood plasma for inhibiting Na,K-ATPase, which indicates the presence of circulating EDF in blood plasma. PMID- 2555932 TI - Small cell carcinoma of the oesophagus: report of three cases and review of published cases. AB - Small cell carcinoma of the oesophagus is a rare tumour with a poor prognosis. This report presents evidence favouring chemotherapy as the primary treatment. PMID- 2555933 TI - Treatment of oesophageal small cell carcinoma by combined chemotherapy and surgical resection: report of two cases and review of published cases. AB - Two patients with primary small cell carcinoma of the oesophagus were treated by multidrug chemotherapy followed by surgical resection. A partial response to chemotherapy was observed in both patients. PMID- 2555934 TI - Isolation of cytomegalovirus from the lung tissue of patients with intrapulmonary neoplasma. AB - Cytomegalovirus in the lung tissue was detected by demonstrating cytopathic effects on culture of fibroblasts. Three clinical features were common to patients in whose lung tissue cytomegalovirus was identified: (1) They had lung cancer. (2) They were on a high dose of prednisolone. (3) There were diffuse shadows on their chest x-ray film. PMID- 2555935 TI - Production of reactive oxygen species due to metabolic activation of butylated hydroxyanisole. AB - The synthetic antioxidant butylated hydroxyanisole (BHA) stimulates superoxide formation in rat liver microsomes up to 10-fold. This stimulation is prevented by the monooxygenase inhibitor metyrapone and does not occur when NADH is consumed instead of NADPH indicating that metabolic activation is required for superoxide production. The BHA metabolite tert-butylhydroquinone (TBHQ) is much more active than BHA in stimulating superoxide production, and the amounts of TBHQ and formaldehyde formed from BHA in microsomes are sufficient to explain the effect of BHA. In buffer and in a xanthine oxidase system, superoxide production by TBHQ also takes place. TBHQ autoxidizes to tert-butylquinone (TBQ) and TBQ exceeds TBHQ by far in its capacity for superoxide production in microsomes. Thus, a 30 fold increase of basal superoxide production is induced by 5 microM TBQ. In rat forestomach, the target organ of BHA carcinogenicity in rodents, stimulation of superoxide production by BHA and more markedly by TBHQ and TBQ is also observed. Excess production of superoxide in microsomes by TBHQ is accompanied by excess production of hydrogen peroxide and of hydroxyl radicals. It is concluded that TBQ undergoes redox cycling leading to an oxidative burst in the presence of enzymes capable of one electron reduction of TBQ and that the BHA metabolite TBHQ enters the redox cycle by autoxidation. No oxygen activating properties can be ascribed to BHA itself. PMID- 2555936 TI - Effect of cytochrome c concentration on the ultrastructural appearance of bovine nasal cartilage proteoglycans. AB - Bovine nasal cartilage proteoglycan monomers were studied by Kleinschmidt and Zahn's molecular spreading technique as modified by Rosenberg et al. By decreasing the cytochrome c concentration in the epiphase to 2 micrograms per 100 microliters we were able on nitrocellulose-coated grids routinely to obtain highly contrasted and well spread proteoglycan monomers with a characteristic brush-like appearance and, sometimes, a clearly distinguishable hyaluronic acid binding region. Previously, a hyaluronic acid binding region has only been observed routinely in spread proteoglycan aggregates, and a brush-like structure of proteoglycan monomers on carbon-coated grids, but with considerably less precision due to the poor contrast of the molecules. Molecular spreading was further improved by decreasing the cytochrome c concentration in the epiphase to less than 2 micrograms per 100 microliters, but contrast was reduced making visualization of molecular details difficult. PMID- 2555937 TI - [The dynamic characteristics of regional muscle blood flow and the activity of mineralization processes in the bony callus with various methods of treating mandibular fractures]. AB - RMBF was comparatively assessed in patients with lower jaw fractures under orthopedic therapy of the osteal suture. To this end, the 99m pertechnetate was injected bilaterally in masseter muscles. The data suggest that double jaw fixation had an adverse effect on RMBF, especially ipsilateral to the fracture. Nevertheless, the mineral metabolism intensity in the callus is higher under fixation as related to the cases surgically sutured. PMID- 2555938 TI - Prevention of primary cytomegalovirus infection in patients with hematologic malignancies by intensive white cell depletion of blood products. AB - The effect of white cell depletion of red cells and platelet concentrates on the transmission of cytomegalovirus (CMV) was studied retrospectively in 150 patients treated intensively for acute leukemia or non-Hodgkin's lymphoma. CMV infection was diagnosed on the basis of IgM and IgG antibody responses to CMV late antigen (CMV-LA). Before cytoreductive therapy for their underlying disease, 59 patients were CMV seronegative and 91 were CMV seropositive. None of the 59 CMV seronegative patients showed persistent seroconversion 2 months after the cytoreductive treatment. The comparison group, consisting of 312 cardiac surgery patients, showed a significantly higher incidence of primary CMV infections: 10 of 86 (11.6%, p = 0.004). Twenty-five percent of the CMV-seronegative patients and controls had transient IgG antibodies to CMV-LA without IgM antibodies, which is indicative of antibodies passively acquired via blood products. These results indicate that white cell-poor blood products carry a very low risk, if any, of CMV transmission. The policy of transfusing white cell-poor blood products provides a useful alternative to the selection of CMV-seronegative donors. PMID- 2555939 TI - Neonatal pretransfusion testing for cytomegalovirus antibody. PMID- 2555940 TI - Site-specific recombination by Tn3 resolvase. AB - Site-specific recombination processes in microbes bring about precise DNA rearrangements which have diverse and important biological functions. The sites and recombinase enzymes used for these processes fall into two distinct families. Here we describe how experiments with one family, exemplified by the resolution system of transposon Tn3, have provided insight into the ways in which DNA and protein interact to bring together distant recombination sites and promote strand exchange. PMID- 2555941 TI - Increasing complexity revealed in regulation of Ca2+ antagonist receptor. PMID- 2555942 TI - Alpha-adrenoceptors in human and animal cerebral arteries: alterations after sympathetic denervation and subarachnoid hemorrhage. AB - Vasospasm of cerebral arteries in patients with subarachnoid hemorrhage frequently presents severe clinical problems resulting from cerebral ischemia, but the pathogenesis of vasospasm is still poorly understood. The contractile response of human cerebral arteries to noradrenaline is larger than the responses in other species. Neurogenic factors controlling brain circulation may play an important role in pathological conditions such as subarachnoid hemorrhage. Motohatsu Fujiwara and colleagues review species variations of alpha adrenoceptors in cerebral arteries and their alterations after surgical sympathectomy. They compare these changes to those occurring in human cerebral arteries following subarachnoid hemorrhage and discuss their relationship to vasospasm. PMID- 2555943 TI - [The DNA content in the cell nuclei of trophoblastic tumors]. AB - The DNA content in cytotrophoblast (CTB) and syncytiotrophoblast (STB) cell nuclei was assayed in tissue sections of 7 hydatidiform moles (HM) and 27 choriocarcinomas (CH). The procedure involved Feulgen's reaction and scanning cytophotometry. The analysis of summarized histograms showed the DNA distribution in CTB cell nuclei, on the one hand, and that in STB, on the other, to differ significantly in both the tumors. The HM studied cases were referred to as two subtypes on the basis of such parameters as modal class value, its ploidy and degree of nuclear poly- and heteroploidy of CTB and STB. These characteristics were used to identify three patterns of CH. A pronounced modal class (2c--4c) was typical of type 1. A wider range of modal class (2c--10c or 4c--8c) was observed in type 2. Type 3 of tumor was characterized by a pronounced polyploidy with the absence of the modal class. The analysis of individual CTB and histograms showed no significant differences between HM and CH with respect to the DNA content. An increase in the share of highly polyploid cells was associated with a shorter survival of patients. PMID- 2555944 TI - [Peptide-activator of cyclic nucleotide phosphodiesterases in the rat and human uterus]. AB - The presence of the cyclic nucleotide phosphodiesterase peptide-activator previously isolated from the calf myometrium was studied in uterine tissue of rats at different functional states (in puberty, estrus and diestrus stages, pregnancy) and in human myometrium and myoma. The peptide isolated from calf myometrium, immature rat myometrium and uterine tissue in myoma caused a 3-4-fold stimulation of the phosphodiesterase activity. PMID- 2555945 TI - [A non-enzymatic method of preparing isolated hepatocytes: assessment of their morpho-functional activity]. AB - A method for isolating hepatocytes using the two-step liver perfusion by chelate containing solutions is described. The hepatocytes obtained are highly inactive for the trypan blue staining test, (not less than 90%), ATP content, the rate of endogenic respiration and its stimulation by succinate, a degree of xenobiotic oxidation. PMID- 2555946 TI - [Stimulation of energy plastic metabolism by phytohemagglutinin in lymphoid organs and adrenal glands]. AB - It has been established that the concentration of nucleic acids, protein, glycogen, ATP, copper, manganese, zinc as well as the activity of cytochromoxidase and succinate dehydrogenase in tissues of the thymus gland and spleen of albino rats a day after stimulation by phytohemagglutinin increase considerably reaching the maximal values three days later. Taking a prolonged term (up to 7 days) after phytohemagglutinin administration it is found that the content and activity of the studied indices in the thymus and spleen tissues lower regularly, but fail reaching normal values even 14 days after stimulation. In this case changes in the test indices in the spleen are less pronounced than in the thymus in all the studied periods. In the tissues of adrenals a tendency of changes in the manganese and copper content is like that in lymphoid organs in all periods after stimulation by phytohemagglutinin; other test indices have an opposite tendency. PMID- 2555947 TI - [Na,K-ATPase activity in the myocardial sarcolemma in ischemia]. AB - The total time-controlled ischemia (up to 45 min) was studied for its effect on the Na,K-ATPase activity, content of nonesterified fatty acids (NEFA) and intensity of lipid peroxidation (LP) in sarcolemmal (SL) preparations and soluble fractions (SF) from the rat and guinea-pig left ventricles. A strong correlation between Na, K-ATPase inhibition and NEFA accumulation was revealed in the SF. On the contrary, changes in the NEFA content and LP level both in SL and SF did not correlate with a decrease in the enzymic activity. Pretreatment with albumin (0.5 mg/ml) induced equally small increase both in the control and in the ischemic SL preparations. It is suggested that the Na,K-ATPase activity during a short period of myocardial ischemia (up to 45 min) may be due to the NEFA accumulation in the cytosolic and/or extracellular space, but not in SL. PMID- 2555949 TI - [The effect of egg incubation temperature on the activity of cytochrome oxidase and anionic ATPase in chicken ontogenesis]. AB - The short-term cooling of hen eggs under incubation was studied for its effect on the dynamics of the activity of cytochrome oxidase (CO) and anion ATPase in the brain and liver of 15- and 20-day hen embryos and 5-day chickens. The temperature fall in the embryonal period was established to stimulate the activity of bicarbonate-dependent ATPase in the brain and to suppress it in the liver tissue. The CO activity was also subjected to similar alterations. PMID- 2555948 TI - [Features of temperature dependence of the Na+,K+-ATPase reaction in normal and tumorous brain tissue]. AB - It has been shown that in the enzyme preparations (EP) from normal brain tissue (NBT) a typical break on the Arrhenius plot appeared at 20-22 degrees C, nH for Na+ and K+ exceeding 1.7 and 1.4, respectively. In EP from tumoural brain tissue (TBT) no break on the Arrhenius plot at 20-22 degrees C was revealed, but it appeared at 27.5 + 30.5 degrees C. The nH for Na+ with Na+,K+-ATPase from TBT was only 0.9, but the cooperative binding of K+ was preserved (nH = 1.3). Electrophoregrams (EP) from TBT showed additional protein bands. The urea and digitonin treatment of EP from NBT induced a break on the Arrhenius plot at 27.5 30.5 degrees C. It is suggested that the break at 27.5-30.5 degrees C is, probably, accompanied by local changes in the conformation of protein components of the enzyme. PMID- 2555950 TI - [The role of guanidine groups of arginine residues of Na, K-ATPase and glutamate dehydrogenase in an interaction with cholesterol]. AB - Cholesterol was studied in experiments in vitro for its effect on the activity of Na, K-ATPase of the synaptic brain membranes of rats and a crystalline preparation of glutamate dehydrogenase from the liver mitochondria of a bull. Cholesterol decreased the activity of the above enzymes. When blocking guanidine groups of arginine residues of Na, K-ATPase and glutamate dehydrogenase the inhibiting action of cholesterol was absent. The obtained data evidence for the possibility of a direct interaction of cholesterol with membrane enzymes as well as for the important significance of guanidine groups of arginine residues of proteins in the process. PMID- 2555951 TI - [Physico-chemical properties of thiamine kinase from Saccharomyces carlsbergensis yeasts]. AB - The amino acid composition and intrinsic fluorescence were studied in thiamine kinase (ES 2.7.6.2) of brewer's yeast. The enzyme molecule is characterized by higher concentrations of amino acids which promote alpha-helix formation of the protein globule, the amount of residues (cysteine, proline) either binding or folding polypeptide chains being considerably high. Amino acids of middle and low hydrophobicities were the most frequent among the amino acid residues with nonpolar R-groups. The value for the protein isoelectric point was 6.21. The eigen pH value and isoionic point were in good agreement with the isoelectric point value and amounted to 6.28. The fluorescence spectrum has a maximum at 328 nm, half-width at 53 nm and a quantum yield at 0.14 nm. The tryptophane residues were located in hydrophobic surroundings, unexposed to anion quenchers and almost unexposed to cation ones. The fluorescence and phosphofluorescence parameters were sensitive to the conformational changes in the molecule. At pH of 5-9 the protein conformation remained unchanged. The temperature rise above 40 degrees C resulted in a disturbance in the nativity of the globule. The elevation of the enzyme concentration from 0.05 to 1 mg/ml increased the polarization degree from 0.115 to 0.194, the quantum yield and the spectrum position remaining unchanged. The results obtained develop knowledge of the equilibrium system of oligomerous forms of thiamine kinase with different catalytic properties. PMID- 2555952 TI - [Static and dynamic pH-dependent components of calcium exchange in the fraction of smooth muscle sarcolemma vesicles]. AB - It is proved that in the fraction of inverted vesicles of the myometrium sarcolemma there are two components of calcium metabolism which depend on the proton concentration in the incubation medium. The first component, a static one, identified under alkalization of the incubation medium from pH 6.0 up to pH 8.0 under equilibrium conditions (Ca2+ concentration inside and outside vesicles is the same) is manifested as an increase of the calcium capacity of vesicles at the expense of Ca2+-binding centres of the inner surface of membrane vesicles. The second component, a dynamic one, is represented as a passive transmembrane flow of Ca2+ outflowing from the vesicles induced by alkalization of the extravesicle space. Alkalization-stimulated Ca2+ release from vesicles is analyzed kinetically. Possible functional role of two components of pH-dependent metabolism of Ca2+ in providing the electrical and pharmacological-mechanical conjugation in the smooth-muscular tissue is under discussion. PMID- 2555953 TI - [Endocrine intestinal low molecular weight factor inhibits Na+,K+- ATPase activity of enterocytes]. AB - The biological activity of the endocrine secretum fraction isolated from the rat duodenum is determined. The fraction with the molecular weight about 3 kDa is found to possess the factor which inhibits the Na+,K+-ATPase activity of enterocytes. It is found that the inhibitory factor secretum depends on the solution which irrigates the duodenum cavity. The possible regulatory role of the intestine inhibitory factor is under discussion. PMID- 2555954 TI - [Side effects in children treated with ACTH in infantile spasm]. PMID- 2555955 TI - [Occurrence of the yeast, Cryptococcus (Cr) neoformans, in pigeon droppings]. AB - Pigeon droppings were collected in Copenhagen and Odense. In samples from pigeon lofts, cryptococci were found in 33% from Copenhagen and in 16% from Odense. All of the species of cryptococci found were Cr. neoformans. These findings are compared with the previous Danish investigations. The frequent occurrence of cryptococci in AIDS patients is mentioned. PMID- 2555956 TI - [Irreversible renal failure during treatment with angiotensin I converting enzyme inhibitor in bilateral renal stenosis]. AB - A case of irreversible renal failure during treatment with enalapril in bilateral renal artery stenosis is described. In the use of converting enzyme inhibitors, caution and monitoring of renal function during treatment is advised. PMID- 2555957 TI - Pathology of infectious diarrhea of infant rats (IDIR) induced by an antigenically distinct rotavirus. AB - Suckling rats were inoculated with a group B rotavirus to determine the progression of the morphologic changes induced in the intestine by this virus. Several changes were observed by light microscopy 1 day after viral inoculation: shortening of small intestinal villi, villous epithelial necrosis, and villous epithelial syncytia. The lesions were most often present in the distal small intestine, although other small intestinal segments were affected to a lesser degree. By day 3 post-inoculation, epithelial necrosis, and syncytia were no longer present; however, the villous epithelium was disorganized and irregularly vacuolated, and intestinal crypt epithelium was hyperplastic. Alterations in villous height to crypt depth ratios were present in portions of the small intestine for the remainder of the 12-day study period. Epithelial syncytia appeared to form by the breakdown of the lateral interdigitating membranes of the absorptive villous epithelium. Viral particles, abundant in the syncytia, appeared to form from amorphous or reticular arrays of viral precursor material. Group B rotaviral antigens, as detected by indirect immunofluorescence, were present in large amounts in the small intestinal villous epithelium only on the first day after viral inoculation. These studies show that two important diagnostic features of group B rotaviral infections of rats, epithelial syncytia and viral antigen as determined by immunofluorescence, are present only on the first day of disease. These findings should be taken into consideration when attempting to diagnose disease induced by this agent. PMID- 2555959 TI - Cowpox in cats. PMID- 2555958 TI - The expression of intermediate filaments in canine mammary glands and their tumors. AB - Monoclonal antibodies specific for different types of intermediate filaments (cytokeratin, vimentin, desmin and neurofilaments) were used to study the histogenesis of canine mammary glands and 57 canine mammary tumors by immunocytochemistry. The intra- and interlobular duct epithelium, acinar, and intralobular myoepithelial cells stained positively for cytokeratin. Peripheral ductal and acinar cells, as well as interstitial cells, stained positively for vimentin. A similar staining pattern was seen in adenomas, complex adenomas, benign mixed tumors, ductular carcinomas, and one myoepithelioma-like tumor. Additionally, cytokeratin positive cells were scattered interstitially in one single adenoma, most complex adenomas, some benign mixed tumors, complex carcinomas, and in the malignant mixed tumors. All stromal cells stained positively for vimentin. The fibrosarcomas were positive only for vimentin, while the following expressed both desmin and cytokeratin: epithelial-like cells in one adenoma, three complex adenomas, the myoepithelioma-like tumor, the single comedo carcinoma, two complex carcinomas, the single lobular carcinoma, one malignant mixed tumor, and three osteosarcomas. Epithelial-like cells in one adenoma, six complex adenomas, two benign mixed tumors, two complex carcinomas, the lobular carcinoma, and the malignant schwannoma stained for neurofilaments. Three tumors, one adenoma, one complex adenoma, and the lobular carcinoma expressed both desmin and neurofilaments in addition to cytokeratin and vimentin. The results show the expression of different types of intermediate filaments and indicate that there might be a stem cell origin in most of the canine mammary tumors. PMID- 2555960 TI - [Sodium transport inhibitor in hypertensives]. AB - The content of sodium transport inhibitor (STI) in the blood and urine was studied in 106 patients with hypertensive disease (HD) in comparison with the clinical course of the disease. STI secretion influenced actively the main parameters of the water-salt homeostasis and hemodynamics in patients with juvenile hypertension and in patients with benign course of HD. In severe progressing course of HD, STI disappears from the blood and urine that may further progression of the disease. PMID- 2555961 TI - [Hypothermia in the treatment of the pain syndromes in diseases of the nerve trunks]. AB - Regional hypothermia was used in the complex treatment of pain syndromes in patients with lumbosacral radiculitis, radiculalgias, neuralgias. Positive results of antalgic therapy in diseases of the peripheral nervous system allow to recommend this method of treatment for wider employment. PMID- 2555962 TI - [A case of insulinoma with neurologic manifestations]. PMID- 2555963 TI - [A study of the effect of thymotropin on immunologic indicators and excretion of virus in women with chronic cytomegalovirus infection]. AB - Thymotropic hormone was given subcutaneously in a dose of 100 mg twice a week for 3 months to 10 women with verified chronic cytomegalovirus infection. After termination of the treatment, in 8 women the virus excretion was decreased and this was accompanied with some improvement in immunological parameters: increased activity of natural killers, enhanced interferon-producing activity of lymphocytes, higher theophylline-resistant lymphocytes, and lower theophylline sensitive T-cells. The above results confirm the association between the activity of cytomegalovirus infection and the state of immunity and allow thymotropic hormone to be recommended for treatment of other groups of patients with cytomegalovirus infection. PMID- 2555964 TI - [The effect of antigen-specific suppression induced by flaviviruses on the formation of an antigen-specific immunologic defect]. AB - The effect of experimental flavivirus infection on the formation of antigen induced immunosuppression in mice was studied. For this purpose, BALB/c mice with asymptomatic infection caused by Langat or West Nile virus were inoculated with supraoptimal doses of sheep erythrocytes and then checked for the capacity to respond again with production of hemolysin-synthesizing cells to the challenge with the optimal dose of the same antigen. The effect of the antigen-induced immunosuppression was found to be enhanced against the background of Langat or West Nile virus infection. The effect was most marked in case of priming with a supraoptimal dose of sheep erythrocytes inoculated at 4-6 days of flavivirus infection. The aggravation of the antigen-specific immunological defect against the background of the infectious process was demonstrated to be associated with the activation by flaviviruses of the T-suppressor link of immunoregulation under conditions of supraoptimal antigenic effect on the host. PMID- 2555965 TI - [Effectiveness of Soviet derivatives of phosphonic acid and their combination with interferon inducers in cell cultures and in a model of herpetic meningoencephalitis in mice]. AB - Nationally synthesized chemopreparations: phosphonoacetic (PAA), phosphonoformic (PFA) acids and acycloguanosine (Acg) exhibit a marked antiherpetic effect in cell cultures and marked protective effect in herpes meningoencephalitis in mice induced by intraperitoneal inoculation of herpes simplex virus type 1 (HSV-1) (43% PFA, 33% PAA, 25% Acg). Both in vitro and in vivo (mouse meningoencephalitis), PFA (its trisodium salt) on the whole proved to be less toxic than PAA but exerted a higher or similar antiherpetic effect. The combined use of pyrophosphate analogues (PAA, PFA) with Acg is more effective that their use separately both in vitro and in herpes meningoencephalitis in mice an produces an additive effect or one similar to it. Systemic inoculation of interferon inducers, lafarine or ridostine, is effective in herpes meningoencephalitis in mice induced by intraperitoneal inoculation of HSV-1 (the protective effect 33% and 26%, respectively). The combined use of ridostine and PAA in herpes meningoencephalitis in mice led to synergistic effect. PMID- 2555966 TI - [Characteristics of electrophoretic types of rotaviruses isolated from children with acute gastroenteritis]. AB - In an analysis of sporadic cases of gastroenteritis in children, rotaviruses were detected in 61 out of 363 specimens examined. Studies of rotavirus isolates by polyacrylamide gel electrophoresis (EP) of RNA revealed 9 different EP types. In one region, during the entire epidemic season one "long" EP type was prevalent. In one patient, a mixture of a "short" and "long" EP types was detected. In another region, "atypical" rotaviruses were found not belonging to the antigenic group A of rotaviruses. PMID- 2555967 TI - [Development and use of immunoenzyme analysis for diagnosing sandfly fever]. PMID- 2555968 TI - [A comparative assessment of serologic methods of determining antigens to poliomyelitis virus]. PMID- 2555969 TI - [Use of the passive hemagglutination reaction for diagnosing rotavirus infections]. PMID- 2555970 TI - [Assessment of diagnostic prospects for a method of immunoenzyme analysis during mass testing of feces of patients for the purpose of detecting rotavirus antigens]. PMID- 2555971 TI - [Chronic nonspecific diarrhea in the infant]. PMID- 2555972 TI - WHO Expert Committee on Drug Dependence. Twenty-sixth Report. PMID- 2555973 TI - [Development of a rational monitoring strategy for workers exposed to tetrachloroethylene]. AB - A 5 year follow-up of 130 workers exposed to tetrachlorethylene (dry cleaning) was performed, including measuring of exposure conditions (personal dosimetry, exposure tests) and clinical investigations. No signs of hepatoxicity of tetrachlorethylene could be detected, but slight derangements of nerval functions may develop in long-term exposed workers. Nerve motor and sensory conduction velocity shows a tendency to diminution. Recommendations are derived for biological monitoring and health surveillance of workers exposed to tetrachlorethylene. PMID- 2555974 TI - [Malignant fibrous histiocytoma of the thyroid]. AB - A report is given on a 46 year-old-man with a malignant fibrous histiocytoma of the thyroid gland. The patient died 4 months after the diagnosis. The autopsy showed progressing tumor mass and metastases in lymphatic nodes, lungs and pleura. PMID- 2555975 TI - [Effect of chronic alcohol drinking and liver cirrhosis on oro-cecal transit time (H2 breath test)]. AB - In a clinical study (1.1.1986-28.2.1988) the oro-cecal transit time of lactulose was studied in 45 patients with liver cirrhosis of different etiologies by hydrogen breath test. The results were compared to the transit time of 16 healthy volunteers. The oro-cecal transit time of patients with alcoholic cirrhosis (means = 154 +/- 29 min.) was significantly increased (P less than or equal to 0.001) while non alcoholic cirrhosis (means = 99 +/- 34 min.) did not differ from the control group (means = 98 +/- 16 min.) As an explanation of the increased oro cecal transit time an alcoholic autonomic neuropathy (- greater than n. vagus) is discussed. PMID- 2555976 TI - [Peracute constrictive, idiopathic pericarditis--a case report of an acute life threatening disease picture]. AB - Generally, idiopathic pericarditis is considered a benign, self-limiting disease. Frequently, the exsudative phase of the disease is followed by a mild form of transitory constriction of the pericardium. The case reported here shows an unusual course of the disease. Shortly after the symptoms of exsudative pericarditis subsided a life-threatening form of pericardial constriction developed within weeks. In case of chronic pericardial constriction perioperative mortality for partial pericardiectomy is not insignificant. This is a result of myocardial damage that is difficult to assess prior to surgery. For that reason a partial pericardiectomy should be attempted as early as possible, even in cases with acute pericardial constriction. PMID- 2555977 TI - Effect of pertussis toxin and the cGMP lowering agent LY83583 on the relaxation induced by nitrates in isolated bovine mesenteric artery. A comparison between glyceryl trinitrate, isosorbide dinitrate and isosorbide 5-mononitrate. AB - The effect of pertussis toxin (PTX) and the cyclic GMP-lowering agent LY83583 on the relaxatory response induced by glyceryl trinitrate (GTN), isosorbide dinitrate (ISDN) and isosorbide-5-mononitrate (ISMN) in bovine mesenteric artery was investigated. Pretreatment with PTX (100 ng/ml; 2h) induced a 100-fold right shift of the concentration-effect curve for GTN, while no effect on the relaxatory response elicited by ISDN and ISMN was seen 10 microM LY83583 markedly reduced the relaxatory effect of all the organic nitroesters. Based on the different sensitivity towards PTX it is suggested that GTN induces vascular smooth muscle relaxation by a partly different mechanism than ISDN and ISMN. However, cGMP seems to play a crucial role in mediating the relaxatory response of all the tested organic nitroesters since LY83583 profoundly suppressed the relaxatory response. PMID- 2555978 TI - The role of nitric oxide formation in organic nitrate-induced vasodilation and organic nitrate tolerance. AB - Isolated rabbit aortic strips (RAS) were contracted submaximally with phenylephrine (PE) and then were incubated with 6.2 x 10(-7) M [3H] glyceryl trinitrate (GTN) (9.98 Ci/mmol) in a 30-s time-course study. GTN-induced relaxation of RAS was monitored and tissue GTN and GDN concentrations were determined by thin-layer chromatography and liquid scintillation spectrometry. There was time-dependent biotransformation of GTN to glyceryl dinitrate (GDN) by the RAS and a time-dependent increase in cyclic GMP content in the RAS. Statistically significant (P less than 0.05) biotransformation of GTN and elevation of cyclic GMP content in the tissue were found at 10 s in RAS, whereas the onset of relaxation occurred at 12 s. During the tissue biotransformation of GTN, there was preferential formation of 1,2-GDN compared with 1,3-GDN, suggesting that it is during the process of conversion of GTN to 1,2-GDN that elevation of cyclic GMP content occurs. The results of this study are consistent with the hypothesis that GTN is a prodrug, such that biotransformation to the active metabolite, nitric oxide (NO), is involved in GTN-induced relaxation of vascular smooth muscle. The data also indicate that the mechanism of GTN biotransformation and GTN-induced activation of guanylate cyclase may be related intimately. Isolated RAS were made tolerant to organic nitrates in vitro by incubation with 5 x 10(-4) M GTN for 1 h. After washout and submaximal contraction with PE, the tissues were incubated with 5 x 10(-7) M [14C]GTN for 2 min.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555979 TI - [Isolated human venous segments as a model for the study of problems of nitrate tolerance]. AB - Concentration-dependent relaxation (6-70%) of segments of human saphenous veins under isometric conditions could be demonstrated with cumulative concentrations of isosorbide dinitrate (ISDN) and Glycerol trinitrate GTN (10(-9)-10(-5) M). Vein segments were obtained during coronary by-pass surgery. Nitrate (GTN) induced relaxation was accompanied by a 2- to 3-fold increase of cyclic GMP content in the vessel walls. However, no change of concentrations in the vessel walls could be determined for the metabolites of prostaglandines: (PG E2, PG F2 alpha, TX B2, 6-keto-PGF1 alpha). Pretreatment of patients with 40 mg ISDN (standard release formulation) 4 times daily for 1 week prior to surgery with the last dose 1 hour before harvesting the vein segments did not influence relaxation. by ISDN. Immersion of vein segments for 1 hour in buffer solution containing 10(-6) M ISDN (= therapeutic concentration) prior to relaxation with cumulative concentrations of ISDN did not influence relaxation either. Induction of in vitro tolerance required ISDN concentrations which exceeded the range achieved under therapeutic conditions: 4.4 x 10(-4) M. This in vitro tolerance could be widely reversed by 10 mM N-Acetylcysteine (NAC) suggesting involvement of sulfhydril (SH) groups. Since tolerance in this experimental model was not seen under concentrations achieved in patients it seems likely that clinical tolerance is caused by activation of counterregulatory forces. PMID- 2555980 TI - [The role of papillomaviruses in precancerous dermatoses and malignant neoplasms of human skin and mucous membranes]. PMID- 2555981 TI - [Morphologic analysis of neuroectodermal elements in mature ovarian teratomas]. AB - Morphologic analysis of neuroectodermal glial tissue was studied in 43 cases of mature cystic teratomas of the ovary. During investigations it was stated that the borders of glial tissue within teratoma, in relation to surrounding mesenchymal structures, are different. Sometimes the glial nests revealed invasive growth in spite of their histologic maturity, while others had insular, polycyclic shapes and sharp borders. These first nests, according to opinion of the author, seem to be potential source of peritoneal infiltration in the cases of peritoneal gliomatosis. Some histologic structures, observed by the author, suggest that during the differentiation of teratoma of the ovary there are morphogenetic movements similar to those known from embryo- or histogenesis, like abortive migration of glial cells to form abortive cerebral cortex. PMID- 2555985 TI - [McAb LC-1 against human lung cancer]. AB - Recently Ge et al reported LAC-122, LAC-210 and LAC-163 McAbs against Human non small cell lung cancer and LSC McAbs against human lung small cell carcinoma. The immunotoxin, composed of McAb LAC-122 conjugated with Ricin A chain has been reported to have the significant cytotoxic effect in vitro on lung adenocarcinoma cell line SPC-A-1 by Tan et al. The LAC-122 alone has no effect on this target cell in the presence of complement from human, rabbit or guinea pig. The tumor associated antigens of human lung cancer have been recognized for many years, but only few reports deal with the common antigens or common epitopes of the lung cancer. From one fusion, 20 hybrids had been observed, all of these culture supernatants could react with target cell by IF. One of them after 4 th cloning, immunoglobulin isotype of the monoclonal antibody thus far obtained belonged to IgM and named to LC-1. Table 1 showed the results of ABC staining of LC-1 with a variety of tumors, normal adult and fetal tissues. From 12 non-small cell lung cancers, including 7 lung adenocarcinoma, 2 lung squamous carcinoma, 3 lung giant cell carcinoma, only one adenocarcinoma gave negative staining. As for 6 small cell lung cancers, all of them showed the positive reaction. It could be also reacted with 11 other tumors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2555984 TI - [The use of commercial immunoenzyme and latex preparations for the demonstration of human rotavirus antigens]. AB - The results of using enzyme immunoassay and latex preparations for the diagnosis of rotavirus gastroenteritis are presented. High effectiveness of the enzyme immunoassay system developed in the USSR with latex diagnostic agents, such as Rotalex (Orion Diagnostica, Finland), Slidex Rota Kit (BioMerieux, France), The Wellcome Rotavirus Latex Kit (Wellcome Foundation Ltd., Great Britain), 48-63% and 21-41% respectively, has been noted. The results of the comparison of the system developed in the USSR with Wellcozyme Rotavirus, an enzyme immunoassay system manufactured by Wellcome Foundation Ltd. (Great Britain), are practically comparable. The results of the block test and the confirmation test used for control indicate that the Soviet preparation is specific. Materials on the practical evaluation of the assay system at health institutions are presented. Good prospects for the use of this system in the diagnosis of rotavirus gastroenteritis, as well as in the realization of epidemiological surveillance on this infection, are substantiated. PMID- 2555982 TI - [The influence of endogenous opioids in hypothalamic amenorrhea. Diagnostic and therapeutic aspects]. AB - We investigated the possibility of induction of ovulation by means of chronic opioid receptor blockade with naloxone. Daily 4 mg were given as an intravenous bolus injection in 4 women suffering from hypothalamic amenorrhea till the 30th day of therapy. A possible maturation of ovarian follicles and a subsequent ovulation should be proved by means of daily determination of LH, FSH, estradiol and progesterone as well as of a sonographic folliculometry. The day prior to and the first day of naloxone treatment we took blood samples every 10 minutes during 4 hours for determination of LH pulse frequency and amplitude. Neither we found any alteration of the basal values of LH, FSH and estradiol, nor we observed a follicular growth. These results lead us to the conclusion to introduce a naloxone stimulation test as a further diagnostic step. In this way opioid mediated hypothalamic ovarian insufficiencies could be registered and a sufficient therapy could be reached. PMID- 2555986 TI - The application of fine needle aspiration cytology in the diagnosis of multiple primary malignant tumors. AB - Four cases with multiple primary malignant tumors are presented. In all cases, fine needle aspiration (FNA) cytologic findings indicated the presence of more than one primary malignancy. In one case, the cytologic examination simultaneously diagnosed two separate primaries. Since FNA cytology can often be used to identify the tumor type, it can be utilized in the identification of many multiple primary malignancies, as these cases show. PMID- 2555983 TI - [Comparative pathomorphologic study of the toxic properties of a corpuscular pertussis vaccine and of a cell-free pertussis preparation]. AB - The comparative study of morphological changes in the body of outbred mice under the action of corpuscular pertussis vaccine and acellular pertussis preparation has been made. The corpuscular vaccine has been shown to produce a pronounced, dynamically increasing toxic effect, thus causing the damage of lymphoid thymic and spleen cells, prolonged interstitial reaction in the lungs, destructive inflammatory process at the site of injection. The acellular pertussis preparation is less toxic, induces less pronounced changes in these organs at the early period of the experiment, stimulates the proliferation of lymphoid cells and lymphoblast transformation. As noted in this study, the damaging action of pertussis vaccine is mainly indicated by pathological phenomena appearing in the organs of the immune system, pulmonary parenchyma and muscular tissue (in the inoculation zone). PMID- 2555987 TI - Significance of mild cervical cytologic atypia in a sexually transmitted disease clinic population. AB - While the clinical management of patients with cytologic atypia consistent with cervical intraepithelial neoplasia (CIN) is well established, the management of so-called nondyskaryotic or benign atypias is controversial. The natural history of such atypias was prospectively studied in 124 patients attending a sexually transmitted disease clinic. The benign atypias were subdivided into different categories according to the cell type involved, and the patients were followed cytologically and colposcopically without cervical biopsy until they reached one of the study end points. Benign atypias often signified the presence of CIN: 17% of the patients had a smear consistent with CIN on the first study visit, and an additional 24% had such a smear by six months of follow-up. After 30 months of follow-up, the overall cumulative rate of biopsy-confirmed CIN 2 to CIN 3 was 13.5%; it was 12.5% among those with two consecutive smears showing benign atypias. Of the ten patients who reached biopsy-confirmed CIN 2 to CIN 3, a cytologic smear consistent with CIN was obtained in less than one year of follow up in all but one case; however, colposcopic evidence of progression was seen in only half of the ten cases, suggesting a sampling error rather than true progression in many cases. Patients with metaplastic cell atypia had a higher progression rate to biopsy-confirmed CIN 2 to CIN 3 (21%) than did those with nonmetaplastic cell atypia (3%). This study provides direct evidence of the potential significance of metaplastic atypia as a marker of CIN. We conclude that patients with mild atypia are often found to have CIN and warrant further investigation by colposcopy and biopsy. PMID- 2555988 TI - Cytologic diagnosis of liver fluke infestation in a patient with subsequently documented cholangiocarcinoma. AB - In a 32-year-old Laotian immigrant who presented with a two-day history of vomiting, abdominal pain and jaundice, ultrasound examination revealed a posthepatic obstruction. Characteristic parasitic ova were present in bile fluid submitted for cytologic evaluation. Subsequent biopsy of the patient's bile duct lesion revealed a coexistent cholangiocarcinoma. The life cycles of Clonorchis sinensis and Ospisthorchis viverrini are reviewed along with the clinical and pathologic complications of infestation by these parasites in humans. The cytologic features of liver fluke infestation are characteristic and should be appreciated, as should the importance of its early diagnosis in the prevention of bile duct neoplasms. PMID- 2555989 TI - Polyomavirus infection versus high-grade bladder carcinoma. The importance of cytologic and comparative morphometric studies of plastic-embedded voided urine sediments. AB - The cytodiagnostic criteria of polyomavirus infection of the urinary tract versus high-grade bladder carcinoma in Cytospin and plastic-embedded preparations of voided urine samples are presented. In Cytospin preparations, the polyomavirus infection and the high-grade bladder carcinoma could not always be distinguished from each other. The diagnosis was facilitated when plastic-embedded specimens were used for cytologic study. On the basis of the comparison of morphometric data from the two types of specimens, it is postulated that the physical properties of the cancer cell nuclei differ from those of the virocytes. PMID- 2555990 TI - Benign neurilemmoma (schwannoma) masquerading as a pleomorphic adenoma of the submandibular salivary gland. AB - A case of solitary benign neurilemmoma (schwannoma) arising in the submandibular region is presented. The tumor was mistaken clinically for an enlarged submandibular salivary gland. Fine needle aspiration cytology made an erroneous diagnosis of a pleomorphic adenoma, predominantly stromal in composition. Histology of the resection specimen resulted in the correct diagnosis of a benign schwannoma. Review of the needle aspirate demonstrated cytologic features that should enable both the correct diagnosis of this neoplasm and its distinction from pleomorphic adenoma, which it mimicked in this location. PMID- 2555991 TI - Cytologic detection of parasites in cervical smears. PMID- 2555992 TI - Effects of lithium on stimulated metabolic parameters in dog thyroid slices. AB - Thyroid abnormalities may develop during chronic lithium therapy for affective disorders. Lithium, like iodide, inhibits TSH stimulation of adenylate cyclase and thyroid hormone release. The present study examined the effect of lithium on stimulation of intrathyroidal intermediary metabolism by several agonists. LiCl (5 mmol/l) did not inhibit basal cAMP, glucose oxidation or 32P incorporation into phospholipids in dog thyroid slices. Although LiCl inhibited TSH stimulation of cAMP, it did not abolish the hormone's effect on cAMP-dependent protein kinase. The stimulation of iodide organification, glucose oxidation or 32P incorporation into phospholipids by TSH, carbachol and phorbol esters was not inhibited by lithium. This is in contrast to the effects of iodide, which inhibited stimulation of glucose oxidation and 32P incorporation into phospholipids by various agonists. Thus, although both lithium and iodide inhibited TSH-stimulated cAMP formation, they act differently on intrathyroidal intermediary metabolism. PMID- 2555993 TI - Adrenal function in HIV infected patients. AB - Since anatomopathological lesions of the adrenal gland have been frequently observed at autopsy in AIDS, we investigated the glucocorticoid function in 63 patients (51 men, 12 women) infected by the human immunodeficiency virus (HIV) in order to determine the incidence and the nature of any adrenocortical abnormalities at various stages of HIV infection. The patients were classified according to the Centers for Disease Control (CDC) recommendations into group II (asymptomatic; N = 13), group III (lymphadenopathy; N = 27) and group IV (clinical manifestations; N = 23). Plasma ACTH and cortisol before and after an exogenous ACTH stimulation test were measured in patients as in 30 age-matched controls. Plasma renin activity and plasma aldosterone before and after ACTH stimulation were also measured in 31 patients (group II: 12; group III: 10; group IV: 9). Compared with controls patients from group II-III had higher levels of ACTH (39.11 +/- 17.01 vs 29.73 +/- 8.53 ng/l; p = 0.003) and basal cortisol (232 +/- 91.2 vs 184.3 +/- 30.9 micrograms/l; p = 0.03). No significant differences were noted between group IV patients and controls as to ACTH and basal and stimulated cortisol levels. Among the 63 patients, only one from group IV had a blunted cortisol response after ACTH stimulation test. Plasma renin activity, and basal and stimulated aldosterone levels in the 3 groups of patients were not different from control values. IN CONCLUSION: 1. Adrenal insufficiency does not seem very frequent in group IV patients and is likely to be a late complication in AIDS. 2. The increased ACTH and basal cortisol levels found in group II and group III patients argue for an early dysregulation of the adrenocortical axis in HIV infection. The exact physiopathological mechanism is not yet known, but an enhanced CRH production by interleukin 1 and/or a direct role of the HIV envelope glycoprotein (gp 120) may explain the high ACTH level in HIV patients. PMID- 2555994 TI - Decreased glucagon binding and glucagon-stimulated lipolysis in adipocytes from streptozotocin-diabetic rats. AB - Adipocytes from streptozotocin-diabetic rats showed a markedly reduced lipolytic response to glucagon concomitant with a 90% or greater decrease in the number of glucagon receptors per cell. In contrast, beta-adrenergic receptors assessed by [3H]dihydroalprenolol binding and lipolysis stimulated by isoproterenol, dibutyryl 3'5'-cyclic AMP and 3-isobutyl-1-methylxanthine were reduced by only 10 25% in diabetic rats compared with controls. Furthermore, quantitative analysis of the relationship between the amount of cell-bound glucagon and the hormone stimulated lipolysis revealed that the function of the remaining 10% of glucagon receptors remained intact in cells from diabetic animals. These findings suggest that the lipolytic cascades, including beta-adrenergic receptors, in adipocytes are not greatly impaired by diabetes, and therefore, the unresponsiveness of these cells to glucagon is mostly due to a marked reduction in the number of glucagon receptors, probably as a result of a down-regulation by postprandial hyperglucagonemia. PMID- 2555995 TI - Sequential immunohistochemical expression of IGF-I and the transferrin receptor in regenerating rat muscle in vivo. AB - The immunohistochemical expression of insulin-like growth factor I and the transferrin receptor was investigated in regenerating rat skeletal muscle. Muscle injury was induced in the extensor digitorum longus muscle in one hind limb by tourniquet ischemia. The regenerating muscles were investigated 3 to 7 days after the injury. Expression of IGF-I and the transferrin receptor was demonstrated in cryostat sections by a double-staining method. It was found that both markers were transiently expressed by the regenerating muscle cells. Expression of IGF-I immunoreactivity preceded that of the transferrin receptor and the staining for IGF-I disappeared 1 to 2 days before the staining for the transferrin receptor. The data are compatible with a regulatory role of IGF-I on the transferrin receptor. PMID- 2555996 TI - Activity of insulin receptor kinase and glycogen synthase in skeletal muscle from patients with chronic renal failure. AB - To examine subcellular mechanisms behind the pathogenesis of peripheral insulin resistance in chronic uremic patients, insulin receptor function and glycogen synthase activity were studied in biopsies of skeletal muscle obtained during renal transplant surgery in 9 non-diabetic uremic patients. The results were compared with values obtained in an age- and sex-matched group of subjects with normal renal function, undergoing surgery for urological or gynecological diseases. The recovery of solubilized, wheat germ agglutinin-purified insulin receptors from skeletal muscle was increased among the uremic patients: 49.3 +/- 6.1 vs 31.4 +/- 2.8 fmol/100 mg muscle in healthy controls (p less than 0.03). Basal as well as insulin-stimulated kinase activities of the insulin receptors, expressed as phosphorylation of the synthetic peptide poly(Glu-Tyr(4:1] were similar. In addition, the maximal activity of the glycogen synthase was enhanced in uremic muscle: 26.6 +/- 2.8 vs 19.5 +/- 1.8 nmol.(mg protein)-1.min-1 (p less than 0.05), whereas the half-maximal activation constant for glucose-6-phosphate was identical in the two groups. Likewise, the muscle glycogen concentrations were similar in the uremic patients and the normal controls. In conclusion, our data suggest that neither impaired insulin receptor function nor a reduced maximal glycogen synthase activity of skeletal muscle are involved in the pathogenesis of the insulin resistance of patients with chronic renal failure. PMID- 2555997 TI - On the nomenclature of the IGF binding proteins. PMID- 2555998 TI - Pick bodies in the locus ceruleus. AB - In classical Pick's disease with typical Pick bodies, inclusions resembling those present in the cerebral cortex are frequently found in the locus ceruleus. In three such cases Pick bodies were studied by light and electron microscopy and compared with Lewy bodies, inclusions more commonly found in this location. In contrast to the situation in the cerebral cortex, nerve cells with multiple Pick bodies were often found in the locus ceruleus, but in other respects definite light and electron microscopic differences between Pick bodies and Lewy bodies were present. Pick bodies were slightly basophilic and never had a central core or a peripheral halo. They were intensely argyrophilic. Differences in immunocytochemical reactions were especially marked with antibodies to tau and to paired helical filaments. Pick bodies displayed an intense reaction with these two antibodies, contrasting with that of Lewy bodies, which either lacked reactivity or reacted in a peripheral band. By electron microscopy the Pick bodies were composed of random filaments with smooth contour, whereas typical Lewy bodies had fuzzy deposits on filaments that radiated from a central core. Pick bodies in the locus ceruleus therefore maintained their immunocytochemical and electron microscopic characteristics and did not take on the character of Lewy bodies. Such differences point to a different pathogenesis and perhaps etiology of these two types of inclusions and attest to the marked difference clinically and pathologically between Pick's and Parkinson's diseases. PMID- 2556001 TI - Papilloma virus infection of the vulva. AB - Analyses of the possible presence of human papilloma virus (HPV) DNA were performed on 84 biopsies from 50 women with clinically and histopathologically suspected HPV infection of the vulva. The colposcopic criteria for inclusion in the study were: acetowhite flat lesions with or without fissures and/or diffuse hyperplastic mucosa with a granulated or filamental surface. Directed biopsies for histopathology and DNA hybridization (dot-blot and Southern-blot) were taken. All women had histopathological evidence of HPV infection. Twelve women had cervical and/or vulvar neoplasia. HPV DNA could be demonstrated in the vulvar biopsies from 24 women; 13 women had HPV 16, 3 had HPV 18, HPV 31 and HPV 33, respectively. In addition, 10 biopsies contained HPV DNA of unknown types. Fifty five percent of granulated and filamental lesions and 43% of flat lesions with or without fissures were positive for HPV DNA. 34 women had one or more of the following symptoms: itching, burning, dyspareunia, discharge, fissures, dryness and dysuria. Papillomavirus vulvitis is a sexually transmitted disease which may cause recalcitrant symptoms and/or concomitant neoplasia. It is thus important to recognize the different symptoms and signs of this disease. PMID- 2555999 TI - New aspects of the pathology of neurodegenerative disorders as revealed by ubiquitin antibodies. AB - Ubiquitin has previously been identified as a component of neuronal inclusions in neurodegenerative disorders. In this investigation, we examined tissue from cases of Alzheimer's disease (AD), Pick's disease, Parkinson's disease (PD), and progressive supranuclear palsy (PSP) to identify previously unrecognized ubiquitinated structures and to assess the evolution of neuronal inclusions. In AD, approximately 60% of neurofibrillary tangles (NFTs) that were stained with an anti-paired helical filaments (PHF) serum were identified by the ubiquitin antibodies. Extracellular NFTs were not labelled with anti-PHF but were unlabelled or weakly labelled with anti-ubiquitin antibodies. In Pick's disease, most Pick bodies were strongly labelled by the ubiquitin antibodies, and in addition some hippocampal CA1 neurones contained granular or strand-like ubiquitin-immunoreactive (IR) inclusions associated with more typical Pick bodies. Typical Lewy bodies in PD cases showed an unlabelled central core with an outer ring intensely labelled by ubiquitin antibodies. Pale bodies in pigmented substantia nigra neurones appeared as large well-defined, rounded structures without an identifiable core or peripheral zone. Some pale bodies were unlabelled by ubiquitin antibodies, but others showed labelling of variable intensity. Pale bodies which were labelled by ubiquitin antibodies tended also to be labelled by BF10, a monoclonal antibody against phosphorylated neurofilaments. We suggest that pale bodies in PD may represent stages in the formation of Lewy bodies. In addition, we observed numerous spindle-shaped ubiquitin-IR swellings of dendrites of pigmented substantia nigra neurones. In contrast to inclusions of AD and Pick's disease, the PHF-positive fibrillary neuronal inclusions of PSP were either unlabelled or only weakly labelled by ubiquitin antibodies. No ubiquitinated structures were seen in neurones from corresponding areas in aged controls. Identification of ubiquitinated proteins in neurodegenerative disorders may provide insights into molecular events associated with cell death. PMID- 2556000 TI - Intracytoplasmic eosinophilic inclusions in the neurons of the central nervous system. AB - This study reports the histological, ultrastructural, and immunocytochemical characteristics of intracytoplasmic eosinophilic inclusion bodies occurring in various types of neurons of the human central nervous system. By light microscopy, the inclusions were brightly eosinophilic, slightly birefringent, and sharply demarcated; they were found in the thalamus in 92% of the cases, in the substantia nigra in 88%, in the locus coeruleus in 45%, and rarely in the spinal cord. Ultrastructurally, the inclusions were composed of assemblies of parallel, alternating dark and light rectilinear profiles. The dark profiles corresponded to thin filaments (microfilaments) that measured 5.5-6.0 nm in diameter. A second set of dense lines crisscrossed the first at right angles. In sections perpendicular to their long axis, the filaments were distributed in a tetragonal lattice pattern in which individual elements occupied the angles of a square. Immunocytochemical preparations for actin were negative. Due to their high rate of occurrence in nonpathological brains, it is thought that the inclusions represent a normal but as yet unidentified cytoplasmic organelle. PMID- 2556002 TI - Risk factors for cataract in Oxfordshire: diabetes, peripheral neuropathy, myopia, glaucoma and diarrhoea. AB - 423 cataract patients and 608 controls between the ages of 50 and 79 were interviewed in a case-control study in Oxfordshire. Diabetes, myopia, glaucoma, peripheral neuropathy and severe diarrhoea were identified as risk factors. The excess risk experienced by females with diabetes was confirmed. The trauma of surgery for glaucoma may be largely responsible for the appearance of glaucoma as a risk factor. Severe diarrhoea has now been identified as a risk factor in England and in India. The risk associated with peripheral neuropathy may indicate a common aetiology at least for some proportions of the two conditions. PMID- 2556003 TI - Fibrous histiocytoma of limbus. AB - The authors report a case of fibrohistiocytoma of the limbus and discuss the clinical, histopathological and immunohistochemical findings concerning this type of lesion, with a comparison of their findings with those reported in the literature. PMID- 2556004 TI - Ultracytochemical demonstration of phospholipids in the surface layer of the guinea pig eustachian tube. AB - The surface layer, especially the extracellular mucous blanket, in the eustachian tube of adult guinea pigs, was preserved very effectively with an intravascular perfusion of twice diluted Karnovsky's fixative following overnight incubation in the same fixative containing 1% tannic acid. Multilamellar bodies were observed in both mucous lining layers of the tube and the epithelial cells of its lining, but neither reticular nor lattice-like ultrastructures could be detected. Mucous blankets on the pharyngeal portion were composed of an electron-dense surface film and a hypophase. The hypophase was marked heterogenous, and it could hardly be observed on the isthmus. The enzymic digestive method using a purified phospholipase A2 was applied to the middle portion of the tube, and the reaction products, probably phospholipids, could be seen both on the surface film of the lining layers and around some of the dark inclusion bodies in the secretory cells. These findings suggest that such phospholipids may play an important role in reducing surface tension of the eustachian tube. PMID- 2556005 TI - Human sarcomatous Wilms' tumor. Characterization with 5H10, a newly established monoclonal antibody. AB - Immunophenotypic features of human sarcomatous Wilms' tumor (SWT) were studied using a newly established mouse monoclonal antibody (5H10, IgG1). 5H10 was produced against CR-SW2, one of several transplanted SWT lines in nude mice, and defines a 200-kDa cell surface protein. The antibody was found to react equally with all subtypes of SWT; clear cell sarcoma of the kidney, malignant rhabdoid tumor of the kidney, and unclassified sarcoma. Furthermore, it reacted equivalently with surgically resected tumors, transplanted tumors in nude mice, and cell lines in vitro. On the other hand, 5H10 was entirely negative for any of the components of nephroblastic Wilms' tumor (NBW). Considering these results, 5H10 appears to recognize the antigen expressed preferentially on SWT, and therefore the subtypes of SWT may be closely related to one another immunophenotypically. In normal human tissues, however, 5H10 only reacted with fetal kidneys, its reactivity being restricted to the lower limbs of S-bodies in both the metanephros and mesonephros. No reactivity was identified in any other tissues including adult kidney. These results indicate that 5H10 detects an oncofetal antigen expressed preferentially in both SWTs and fetal kidney and that the histogenesis of SWT should be considered in connection with nephrogenesis. PMID- 2556006 TI - Recent development in hormone research. AB - A classical distinction between endocrine cells and neurons cannot be accepted without exception. This dichotomy was first challenged by the concept of neurosecretion. Recent observations indicate that hormone synthesis takes place in many extraendocrine tissues since the gene expression for prohormone synthesis seems to be common for all eukaryotes although the secretion of biological active hormone products is limited by posttranslational processing for differentiated cells. Increasing number of data support the view that regulation of pituitary hormone secretion is under multifactorial control in addition to specific signaling molecular effects of hormone-releasing hormones. Such modulators are co secreted messengers from hypothalamic sources or co-functioning at the pituitary cell level. Multichannel regulation of pituitary tropic hormones appears to be important for understanding the interactions of pharmacological agents with pituitary hormone release, on the one hand, and the modulation of hormone release in pathological conditions, on the other hand. Perinatal transient hazards may induce permanent alterations in adaptive behavior when tested in adult age. Corticosteroid-induced deviation of avoidance behavioral reactions may be opposed by simultaneous administration of ACTH-like peptides. These observations revealed that a balance of the glucocorticoids and ACTH-like peptides in perinatal period basically determine the adaptative reaction of animals in adult age. Immune system may be called as a mobile brain since its tremendous information capacity and its responsiveness to alterations of chemical environmental signals. Recent data support the view that there is a bidirectional communication between the neuro-endocrine adaptational axis and the immune system. Stress hormones can alter the immune response and mononuclear cells produce factors that change the neuroendocrine regulation. In addition to these, prohormones are synthesized in mononuclear cells that may be involved in regulation of signalization between cells and in activation of endocrine system and brain functions. PMID- 2556007 TI - Biochemical compartmentation of fish tissues: chronic toxicity of mercuric nitrate on visceral phosphomonoesterases in Channa punctatus (Bloch). AB - Effect of sub-lethal dose of mercuric nitrate was studied in anterior, middle and posterior regions of kidney, anterior and posterior regions of left and right liver lobes, cephalic, thoracic and caudal regions of muscle and left and right gill tissues of C. punctatus in relation to acid and alkaline phosphatase under chronic studies. Middle region of kidney registered maximum rise and fall of acid and alkaline phosphatase, respectively. Right lobe of liver showed more rise and fall of acid and alkaline phosphatase activity respectively. Similarly, left gill is more pronounced than the right one. The observed enzymatic variations were discussed in relation to the biochemical constituents and physiological coordination rendered by these tissues. PMID- 2556008 TI - Basal and stimulated LHRH receptor sites in the pituitary of aging female mice. AB - We studied the regulation of pituitary luteinizing hormone-releasing hormone (LHRH) receptors in female mice aged 4-8 months (young), 10-14 months (middle aged) and 15-18 months (old). In intact animals the mean +/- SE pituitary content of LHRH receptor sites ranged from 34.4 +/- 6.3 fmoles to 41.5 +/- 5.0 fmoles and remained constant throughout aging. Similarly, receptor affinity for LHRH was unaltered (range 1.0-4.0 X 10(9) M-1) by age. Ovariectomy (ovx) resulted in a 30 50% decrease in pituitary LHRH receptor sites (p less than 0.05) and the pattern of LHRH receptor fall after ovx was similar in all three age groups. Administration of estrogens (E2) at either a low (0.2 mg) or a high (2.0 mg) concentration to ovx mice prevented this fall in LHRH receptor sites and, in all three age groups, increased pituitary values to levels comparable to those measured in intact animals. We conclude that there are no significant differences in the basal and stimulated number or affinity of pituitary LHRH receptors between young, middle-aged and aged female mice. PMID- 2556009 TI - Scintigraphic study of cerebrospinal fluid shunts. AB - This paper deals with experience gained in the study of patency of different types of shunt. Results of subsequent surgery are considered. Findings of 79 scintigraphic studies with radiotechnetium on 68 patients are presented. The method is simple, quick, easy to reproduce and means only moderate radiation load for the patient. Results obtained suggest that this study is a reliable diagnostic tool which can be used in a routine way. PMID- 2556010 TI - CDC releases updated guidelines for STD treatment. PMID- 2556011 TI - Aggressive insulinoma with bone metastases. AB - This is the first report of the development of bone metastases in a patient with an insulinoma. Following the development of bone metastases, further chemotherapy with etoposide (VP-16) and cis-platinum was not successful and the patient's condition was rapidly fatal. This case demonstrates the unusually aggressive course and poor response to currently available chemotherapeutic agents of a malignant insulinoma. The clinical and diagnostic features of insulinomas are reviewed. PMID- 2556012 TI - Phase II study of idarubicin in extensive-disease non-small-cell lung cancer. AB - Fourteen patients with extensive-disease non-small-cell lung cancer (E-NSCLC) were treated with oral 4-demethoxydaunorubicin (idarubicin, 4DMDR) at a dosage of 10 mg/m2/day x 5 days every 3 weeks. The median cumulative dose was 110 mg/m2 (range: 50-1,100). Two patients had stable disease for 12 and 56 weeks, respectively, one patient had failed to respond to a doxorubicin hydrochloride (Adriamycin)-containing regimen, and one had had no prior therapy. Twelve of the 14 patients had prior radiotherapy, chemotherapy, or both. Median survival for this heavily treated group was 16 weeks. Myelosuppression was minimal. Nausea and vomiting occurred in 44% of all courses. No cardiac toxicity and no decrease in cardiac ejection fraction was observed. We conclude that 4DMDR is ineffective in heavily treated E-NSCLC patients. However, the drug's activity in untreated patients is unknown. Further study of 4DMDR is indicated in patients who have had no prior chemotherapy or radiotherapy, with routine administration of antiemetic drugs along with pharmacokinetic studies. PMID- 2556013 TI - 5-Fluorouracil infusion and mitomycin combination chemotherapy in the management of patients with advanced non-small-cell lung cancer. AB - We have carried out a phase II study evaluating the activity of a 5-fluorouracil drug combination in patients with advanced non-small-cell lung cancer. Patients were given 60 mg/m2 of methotrexate i.v. on day 1. On day 2, 750 mg/m2 of 5 fluorouracil was administered as a 24-h infusion daily for 3 days. Also on day 3, 10 mg/m2 of mitomycin was given i.v. along with folinic acid. Folinic acid was started on day 3 initially at a dose of 25 mg/m2 intravenously every 6 h for three doses, followed by a 2-h infusion of 200 mg/m2 daily on days 3 and 4. Therapy was repeated every 28 days. Fourteen of 35 patients (40%) experienced a partial response to chemotherapy. The median survival of the entire group was 19 weeks. Mucositis was a common side effect but severe leukopenia, anemia, renal insufficiency, and skin ulceration were rare. This study demonstrated that 5 fluorouracil infusion therapy has activity in advanced non-small-cell lung cancer but the responses are not durable. Further studies evaluating differing dose schedules and alternate 5-fluorouracil infusion-based drug combinations seems warranted. PMID- 2556014 TI - Electrochemical treatment of cancer. I: Variable response to anodic and cathodic fields. AB - Percutaneous placement of an intraneoplasmic electrode in pulmonary metastases of three patients with extrathoracic primary cancers permitted electrochemical treatment of these lesions. These reacted variably to anodic and cathodic electrodes. One breast cancer metastasis disappeared after treatment with the anodic field. A small metastasis from a cancer of the urinary bladder was found to be resistant to both anodic and cathodic fields despite large doses of current. Two large metastases of about equal size from the leiomyosarcoma of the uterus showed progression of the cranial anodic neoplasm but regression of the caudal cathodic neoplasm. At reversed polarity the cathodic neoplasm showed a tendency to regress and the caudal neoplasm continued to disappear. It appears that different neoplasms may show a variable response to anodic or cathodic fields or may show poor sensitivity to both fields. PMID- 2556015 TI - Adenoid cystic carcinoma. A clinicopathologic study with flow cytometric analysis. AB - Forty-five patients with adenoid cystic carcinoma (ACC) of salivary glands were retrospectively studied to determine the prognostic use of DNA ploidy analysis compared with clinicopathologic features. Nuclear suspensions were prepared from 37 of these tumors by the Hedley technique on paraffin-embedded material. The DNA content was analyzed by flow cytometry after propidium iodide staining. Thirty five tumors were diploid and 2 were tetraploid. Mean survival was 117 and 52 months for the diploid and tetraploid patients, respectively. The median S-phase fraction (Spf) of the 35 diploid tumors was 4.45%. Of the 17 diploid patients who died of disease, there were 11 tumors with high Spf (greater than 4.45%) and 6 tumors with a low Spf (P less than 0.05 chi-square test). The presence of more than 30% of a solid pattern in the tumor was strongly associated with poor median survival in Kaplan-Meier survival curve analysis (P less than 0.05 log rank test). Grade, stage, recurrence, and metastases were also found to be important prognostic factors. Because few tumors were nondiploid, these results suggest that S-phase fraction analysis may be a more useful prognostic indicator than ploidy classification. PMID- 2556016 TI - Human immunodeficiency virus (HIV) and Epstein-Barr virus (EBV) antigens and genome in lymph nodes of HIV-positive patients affected by persistent generalized lymphadenopathy (PGL). AB - The presence of human immunodeficiency virus (HIV) and Epstein-Barr virus (EBV) antigens and genome has been investigated in 50 lymph nodes involved by persistent generalized lymphadenopathy (PGL). All the patients were HIV infected and most of them (42 of 50) also had anti-EBV serum antibodies. At lymph node level, HIV and EBV antigens were studied by immunohistochemistry using monoclonal antibodies directed against viral core proteins. The HIV p24 protein was detected in 43 of 50 lymph nodes within the B-cell germinal centers with a reticular pattern. Few cells with positive results for EBV antigens were found in only 2 of 50 lymph nodes. These rare EBV-positive centrocyte-like cells were mainly located in the germinal centers. The presence of HIV and EBV genome was also studied in lymph nodes involved by PGL, with the use of in situ and Southern blot hybridization. A positive reaction for HIV genome was detected in only 1 of 14 lymph nodes with the Southern blot hybridization, and the presence of EBV genome was never demonstrated in these lymph nodes with the use of both in situ and Southern blot hybridization. The expression of EBV antigens and genome was also investigated in the peripheral blood of 15 patients with PGL in which cells with positive results for EBV antigens were detected in a single case with a frequency of 1 X 10(-4). No evidence of EBV genome was found with the use of the in situ hybridization. These results suggest that EBV is not present in lymph nodes during the PGL phase and that its possible implication in the pathogenesis of acquired immune deficiency syndrome (AIDS)-associated lymphoma might be a late event. PMID- 2556017 TI - Comparative features of ductal carcinoma in situ and infiltrating ductal carcinoma of the breast on fine-needle aspiration biopsy. AB - To evaluate the usefulness of fine-needle aspiration biopsy of the breast in separating ductal carcinoma in situ (DCIS) from infiltrating ductal carcinoma, the authors reviewed 16 preoperative fine-needle aspiration biopsies from biopsy proven exclusive DCIS and 39 fine-needle aspiration biopsies from infiltrating ductal carcinomas with or without an in situ component. Seven (44%) of the DCIS and eight (21%) of the infiltrating ductal carcinomas had inadequate material for diagnosis on the aspiration biopsy. Five (32%) of the DCIS and 29 (74%) of the infiltrating ductal carcinomas caused suspicion or had positive results for malignancy. Four (25%) of the DCIS and two (5%) of the infiltrating ductal carcinomas showed atypical cells. Morphologic features of the atypical or malignant cells in the adequate specimens from these two lesions were similar except that the cells from the infiltrating ductal carcinomas showed more irregular nuclear spacing (94% vs. 44%, P less than 0.01) and more pronounced nuclear overlapping (65% vs. 33%) than those from the DCIS. In addition, the fine needle aspiration biopsies of the DCIS tended to be hypocellular (less than 10 cells/10X) (44% vs. 6.5%, P less than 0.05) and to contain benign epithelial cells (22% vs. 6.5%) and macrophages (33% vs. 13%). Although the suspicion of DCIS might be raised when hypocellularity, benign epithelial cells, and macrophages are noted in a fine-needle aspiration biopsy of the breast that has positive results or causes suspicion for malignancy, fine-needle aspiration biopsy cannot be relied upon to distinguish DCIS from infiltrating ductal carcinoma. PMID- 2556018 TI - Bacterial infections in the acquired immune deficiency syndrome. Clinicopathologic correlations in a series of autopsy cases. AB - In a group of 46 adult patients with acquired immunodeficiency syndrome (AIDS) who came to autopsy in 1983-1987, the authors found that 38 (83%) had bacterial (nonmycobacterial) infections some time during the course of their illness, compared with 34 (74%) who had parasitic infections, 31 (67%) who had viral infections, 28 (61%) who had fungal infections, and 12 (26%) who had mycobacterial infections. Twenty-five of these patients (54%) had Staphylococcus aureus infections, compared with 7 (15%) who had Pseudomonas aeruginosa infections and 6 (13%) who had enterococcal infections. Overall, undiagnosed infections or malignancies were found in 48%, 22 of the 46 autopsies, including 12 cases of undiagnosed bacterial infections, 8 of these due to S. aureus. These results suggest that bacterial infections in general, and S. aureus infections in particular, are important causes of morbidity and mortality in patients with AIDS. PMID- 2556020 TI - Lymphoepithelial cystic lesion related to adenocarcinoma in the mediastinum. AB - The authors report here a case of lymphoepithelial cystic lesion (LECL) of unknown origin in the mediastinum, which is closely related to a signet-ring cell adenocarcinoma. A 73-year-old man presented with a mass as revealed on a chest x ray. During surgical operation, a solid, well-circumscribed and encapsulated 9 X 9 X 8 cm tumor was isolated from the right anterior mediastinum. This tumor had neither undergone metastasis nor invaded into the surrounding tissue and lymph nodes. Light microscopy revealed the tumor to be a signet-ring cell adenocarcinoma. Clinically, the neoplasm was coupled with an elevation in serum CEA level, which promptly returned to normal values following surgical removal. Immunohistochemistry pointed out that the majority of neoplastic cells stained positive for CEA. In addition, LECL of unknown origin was distinguished at the periphery of the tumor. LECL was characterized by microcysts which were lined by columnar epithelial and surrounded by lymphoid tissue with germinal centers. Transition between the neoplastic cells and benign epithelial cells of LECL was evident, showing that the previously mentioned mediastinal adenocarcinoma may be derived from the epithelial elements of LECL. This paper discusses the histogenesis of LECL. PMID- 2556019 TI - The mean leukocyte myeloperoxidase index in hematological patients. AB - A Technicon H-1 hematologic analyzer was used to measure the mean leukocyte myeloperoxidase (MPX) in 160 patients seen in a hematology clinic. The normal range was -15 to +10, which included 95% of 300 consecutive hospitalized patients. No abnormalities in the MPX were found in 35 patients with beta thalassemia minor, 8 with iron deficiency, 14 with myeloproliferative disorders, 17 with autoimmune disorders, and 37 patients with lymphoma in complete remission. On the other hand 36% (10/28) of lymphoma patients with active disease either at diagnosis or relapse had a MPX of greater than 10 compared to only 2.3% (7/300) in hospitalized patients (P less than 0.001). Increased levels of MPX were found primarily in patients with non-Hodgkin's lymphoma (NHL) of intermediate or high grades, or Hodgkin's disease [56% (9/16) compared to only 8.3% (1/12) in those with low grade NHLs, P less than 0.05]. The MPX levels returned to normal after successful treatment. Of the various chemotherapeutic agents used, only hydroxyurea led to a consistent elevation of the MPX. The authors conclude that MPX is commonly increased in patients with lymphoma and in those receiving hydroxyurea. Further studies are required to determine if the MPX is a sensitive test for relapse in patients with lymphomas who had an elevated pretreatment value. PMID- 2556021 TI - Malacoplakia of the thyroid gland. AB - A case of malacoplakia of the thyroid gland is described in a 50-year-old Japanese woman. This lesion clinically mimicked a malignant neoplasm, and the true diagnosis of malacoplakia was made only after histologic examination; light microscopy revealed a granulomatous nodule with an accumulation of von Hansemann's histiocytes containing PAS-positive and von Kossa's-positive intracytoplasmic and extracytoplasmic inclusions known as Michaelis-Gutmann bodies. There were some foci consisting of neoplasm-like or hyperplastic residual follicles within the lesion. Electron microscopically, a small number of bacilliform organisms were demonstrated within the lesion. X-ray microanalysis of Michaelis-Gutmann bodies was performed and revealed the presence of phosphorus, calcium, iron, and chloride. It is suggested that the malacoplakic lesion may be associated with the hyperplastic or neoplastic follicular lesion, and bacterial infection could be important in the causation of malacoplakia of the thyroid gland. PMID- 2556022 TI - Endocrine cell hyperplasia of the uterine cervix. A precursor of neuroendocrine carcinoma of the cervix? AB - A 33-year-old woman who presented with vaginal bleeding was diagnosed to have neuroendocrine small cell carcinoma based on cervical smear and biopsy. Hysterectomy was performed, and a tumor measuring 5.5 X 2 mm was found at the squamocolumnar junction of the uterine cervix. In the immediate vicinity of the tumor, there was proliferation of cytologically benign endocrine cells in the normal endocervical glands and in the glands showing intraepithelial glandular neoplasia. Both the hyperplastic endocrine cells and the invasive tumor cells showed argyrophilia and immunostaining for neuron-specific enolase, neurofilament, and chromogranin. The topographical relationship suggests that endocrine cell hyperplasia may represent a precursor of neuroendocrine carcinoma of the cervix. PMID- 2556023 TI - High prevalence rate of human papillomavirus infection and association with abnormal papanicolaou smears in sexually active adolescents. AB - Human papillomaviruses (HPVs) are associated with neoplastic and malignant lesions of the uterine cervix. Cervical neoplasia is associated with onset of sexual activity at an early age. Therefore, this study sought to define the prevalence rates of HPV infection and cytologic abnormalities in adolescents. Sexually active females 13 to 21 years of age undergoing routine cervical cytologic screening were evaluated in the adolescent clinic of an urban hospital. Cells collected by cervicovaginal lavage from 249 subjects were analyzed for HPV DNA by Southern blot hybridization with probes for HPV types 6/11, 16, and 18. The HPV DNA was detected in 95 (38.2%) of 249 patients. Teenagers between the ages of 13 and 18 years with multiple lifetime sexual partners were at higher risk for HPV infection (38/71 [54%]) compared with patients of the same age who had only a single partner (25/74 or [34%]). Twenty (8.3%) of 241 patients had abnormal Papanicolaou smears with atypia, koilocytosis, or low-grade cervical intraepithelial neoplasia. Cytologic abnormalities were detected in 16 (17%) of 94 adolescents with HPV present, but in only 4 (2.7%) of 147 of the uninfected patients. Thus, HPV emerged as a common pathogen in female adolescents, and infected patients are at increased risk for cervical epithelial abnormalities. PMID- 2556024 TI - Study of virus isolation from pharyngeal swabs in children with varicella. AB - We performed virus isolations from the pharyngeal swabs in 117 children with varicella who were aged from 22 days to 15 years and 70 healthy children who were aged from 3 months to 15 years, by using human embryonic lung cell cultures. Viral isolates were confirmed by an indirect immunofluorescence method or by neutralization with well-characterized antibodies. Five varicella-zoster virus isolates (4.3%), 23 cytomegalovirus isolates (19.7%), five herpes simplex virus isolates (4.3%), and one respiratory syncytial virus isolate (0.9%) were found in the patients with varicella. Ten cytomegalovirus isolates (14.3%), two herpes simplex virus isolates (2.9%), one respiratory syncytial virus isolate (1.4%), and one poliovirus isolate (1.4%) were found in the swabs of the healthy control children. The varicella-zoster virus isolation rate from the pharyngeal swabs in children with varicella was low as compared with the rate from those pharyngeal swabs in the children with cytomegalovirus and herpes simplex virus. No varicella zoster virus isolates could be found in the swabbed materials after filtration (0.45 microns). On the other hand, cytomegalovirus and herpes simplex virus could be isolated from the filtrated swabs, as well as from the unfiltrated swabs. The method of testing by filtration could have affected the results. PMID- 2556025 TI - Transmission of chickenpox in a school setting prior to the observed exanthem. AB - An epidemic of chickenpox in a class is described. Four children were able to transmit infection prior to the time their rash was observed by their parents. A fifth child was known to have attended school while he had localized varicelliform lesions that were present for 2 days prior to the appearance of the generalized exanthem. It cannot be ascertained whether some of the other children may have had similar lesions that were not observed at the time they attended school. The first classroom case was observed prospectively so that the time of rash was fairly well established. Although he apparently transmitted infection prior to the onset of rash, virus could not be isolated from the respiratory secretion of this child on the day he presumably infected his classmate. Virus was not found in his respiratory secretions or those obtained from his two siblings before or after the onset of rash, although it was recovered from vesicular fluid. PMID- 2556026 TI - An epidemic of acute hemorrhagic conjunctivitis in American Samoa caused by coxsackievirus A24 variant. AB - Between May 25 and July 5, 1986, an epidemic of acute hemorrhagic conjunctivitis affected an estimated 47% of the population on American Samoa. Coxsackievirus A24 variant was isolated from 18 of 22 patients. This is the first documented outbreak of acute hemorrhagic conjunctivitis due to coxsackievirus A24 variant outside of Southeast Asia and the Indian subcontinent. When this outbreak was compared with an outbreak on the island in 1981-1982 caused by enterovirus 70, conjunctival hemorrhage or injection and the severity of hemorrhage were less prevalent among cases in 1986, while upper respiratory and systemic symptoms were more common. Residents of traditional housing had significantly higher attack rates (48%) than residents of government housing (23%). Serum specimens collected from the residents of Samoa in 1985, before the outbreak, unexpectedly revealed the presence of neutralizing antibodies against coxsackievirus A24 variant. The presence of these antibodies correlated with protection against coxsackievirus A24 variant infection in this outbreak. PMID- 2556027 TI - Assignment of the gene coding for the alpha-subunit of prolyl 4-hydroxylase to human chromosome region 10q21.3-23.1. AB - Prolyl 4-hydroxylase, an alpha 2 beta 2 tetramer, catalyzes the formation of 4 hydroxyproline in collagens by the hydroxylation of proline residues in peptide linkages and plays a crucial role in the synthesis of these proteins. The gene for the beta-subunit of prolyl 4-hydroxylase has recently been mapped to the long arm of human chromosome 17, at band 17q25. We report here chromosomal localization of the gene for the catalytically and regulatorily important alpha subunit of human prolyl 4-hydroxylase. Analysis of 24 rodent x human cell hybrids by Southern blotting with cDNA probes for the human alpha-subunit indicated complete cosegregation of the gene for the alpha-subunit with human chromosome 10. A cell hybrid containing only part of chromosome 10 mapped the gene to 10q11- --qter. In situ hybridization mapped the gene to 10q21.3-23.1. The gene for the alpha-subunit is thus not physically linked to that for the beta-subunit of the enzyme. PMID- 2556028 TI - Cancer risks associated with 10 inorganic dusts: results from a case-control study in Montreal. AB - A multicancer site, multifactor case-control study was undertaken to generate hypotheses about possible occupational carcinogens. Probing interviews were carried out with eligible cases, comprising all incident cases of 20 types of cancer who were male, aged 35-70 years, and a resident in Montreal. The interview was designed to obtain detailed lifetime job histories and information on potential confounders. Each job history was reviewed by a team of chemists and industrial hygienists who translated it into a history of occupational exposures. These occupational exposures were then analyzed as potential risk factors in relation to the sites of cancer included; 3,726 cases were interviewed. For each site of cancer analyzed, controls were selected from among the other sites in the study. This report concerns the associations between the 12 main types of cancer in our series and 10 inorganic dusts that are found mainly in construction and metal industries. All site-exposure combinations were investigated. After intensive control for confounding, nonadenocarcinoma (NAC) of the lung was associated with long duration-high level exposure to silica (odds ratio [OR] = 1.4), excavation dust (OR = 1.9), concrete dust (OR = 2.5), abrasives dust (OR = 1.4), and alumina (OR = 1.5). It was difficult to disentangle the relative effects of those substances, and confounding among them was a distinct possibility. Although residual confounding by some uncontrolled factors may explain the elevated ORs, the results were compatible with the hypothesis of a nonspecific relation between NAC of the lung and respirable inorganic dusts as a class. Other associations that remained suggestive after in-depth analysis were silica and stomach cancer (OR = 1.2) and concrete dust and lymphoma (OR = 2.9). PMID- 2556029 TI - Preoperative lateralization of pituitary microadenomas by petrosal sinus sampling: utility in two patients with non-ACTH-secreting tumors. PMID- 2556030 TI - The use of color Doppler imaging for prenatal diagnosis of umbilical cord anomalies: report of three cases. AB - Color Doppler imaging could easily and rapidly detect the absence of an umbilical artery, despite the presence of oligohydramnios in one case and multiple cord loops involving a normal cord in the other case. In the third case, which on gray scale imaging appeared as an indeterminate cystic structure of the cord, color Doppler imaging demonstrated a complex abnormal vascular pattern suggestive of an angiomyxoma. PMID- 2556031 TI - Fetal intermediate lobe is stimulated by parturition. AB - The fetal pituitary gland secretes beta-endorphin in blood in response to delivery. However, other forms of endorphin have recently been observed in the fetal pituitary, such as N-acetyl-beta-endorphin, which is devoid of opiate activity, and a desacetylated form of alpha-melanocyte-stimulating hormone. Both endorphins originate in the pituitary intermediate lobe. The sensitivity of this lobe to labor stress was assessed by the evaluation of beta-endorphin, N-acetyl beta-endorphin, melanocyte-stimulating hormone, and desacetylated alpha melanocyte-stimulating hormone in maternal plasma and cord blood in 11 cases of vaginal delivery and 10 cases of elective cesarean section without labor. Plasma peptide levels were determined by specific radioimmunoassays after extraction on Sep-Pak C-18 cartridges and high-performance liquid chromatography fractionation. Cord blood samples of infants delivered vaginally showed higher beta-endorphin (8.5 +/- 1.6 pmol/L, mean +/- SE) and desacetylated alpha-melanocyte-stimulating hormone (13.6 +/- 3.2 pmol/L) levels than those delivered by elective cesarean section (3.7 +/- 0.8 and 4.2 +/- 1.1 pmol/L, for beta-endorphin and desacetylated alpha-melanocyte-stimulating hormone, respectively). N-acetyl-beta-endorphin and alpha-melanocyte-stimulating hormone levels do not differ in relation to the mode of delivery. In maternal circulation beta-endorphin levels were higher in those delivered vaginally (5.2 pm 1) than in women who had cesarean sections (2.5 +/- 0.5 pmol/L), whereas no changes were found for the other peptides. In vaginal deliveries, the level of desacetylated alpha-melanocyte-stimulating hormone was higher in cord blood (13.6 +/- 3.2 pmol/L) than in maternal plasma (6.5 +/- 3 pmol/L); there were no significant differences with regard to the other peptides. Fetal and maternal levels of all the peptides were similar in cases of cesarean section. We conclude that parturition activates proopiomelanocortin peptide release from both the anterior and the intermediate pituitary lobe and that the fetus secretes the appropriate beta-endorphin molecule, that is, the peptide able to bind opiate receptors. Concomitant secretion of desacetylated alpha-melanocyte stimulating hormone may occur with adrenal androgen activation at birth. PMID- 2556032 TI - Presence of human papillomavirus deoxyribonucleic acid. PMID- 2556033 TI - Insulin-like growth factor I stimulates Na-dependent Pi transport in cultured kidney cells. AB - The effect of recombinant insulin-like growth factor I (IGF-I/somatomedin C) on the transport of inorganic phosphate (Pi) was studied in cultured kidney epithelia. In opossum kidney (OK) epithelia, IGF-I (5 x 10(-10) to 10(-7) M) induced a dose-related stimulation of the Na-dependent Pi transport (NaPiT). A maximal response was observed at 10(-7) M (IGF-I 1.64 +/- 0.12; vehicle 0.90 +/- 0.02 nmol.mg protein-1. 4 min-1, P less than 0.001). Kinetic analysis of the stimulatory effect of IGF-I on NaPiT indicated an increase in Vmax and no change in Km. Insulin also stimulated NaPiT in OK epithelia but only at concentrations 20-40 times higher than IGF-I. The effect of IGF-I on Pi transport was detectable in less than 30 min with a maximal response occurring after 4-5 h. It was selective for NaPiT, since the Na-dependent alanine transport was not affected by IGF-I. Inhibition of protein synthesis by either cycloheximide or cordycepin markedly attenuated the stimulatory effect of IGF-I on NaPiT. The cellular adenosine 3',5'-cyclic monophosphate content was not modified by the growth factor. In conclusion, these data indicate that IGF-I increases NaPiT selectively through a mechanism that involves de novo protein synthesis. These observations suggest that growth and growth hormone-related stimulation of renal Pi transport could be mediated by IGF-I. PMID- 2556034 TI - Heterogeneous response of cytoplasmic free Ca2+ in proximal convoluted and straight tubule cells in primary culture. AB - Cytoplasmic free calcium [( Ca2+]c) plays an important role in cellular signal transduction in response to hormones. The development of microfluorometric digital image analysis makes it possible to obtain an image of a calcium indicator dye, fura-2, in a single living cell. Subconfluent primary cultured cells from isolated proximal convoluted and straight tubules were analyzed for [Ca2+]c. The basal level of [Ca2+]c was 92.0 +/- 0.82 nM in convoluted and 75.0 +/- 4.7 nM in straight tubule cells. Exposure of convoluted tubule cells to parathyroid hormone (10(-7) M) elicited a 1.4-fold transient rise in [Ca2+]c. Treatment of straight tubule cells with calcitonin (0.1 U/ml), on the other hand, evoked a 1.6-fold transient increase in [Ca2+]c. [Ca2+]c in proximal straight tubule cells did not respond to parathyroid hormone, nor did that in convoluted tubule cells to calcitonin. [Ca2+]c peaks were diminished in straight tubule cells by repeated additions of calcitonin at 5-min intervals. These hormonal responses were abolished by perfusing the cells with calcium-free solution and were not mimicked by perfusion with adenosine 3',5'-cyclic monophosphate (10(-4) M) or with forskolin (5 X 10(-5) M). The results suggest that the [Ca2+]c response to the selected peptide hormones is different in proximal convoluted and straight tubule cells in primary culture. PMID- 2556035 TI - Modulation of osteoblast function by prostaglandins. AB - The naturally occurring prostaglandins (PGs) were studied with respect to their abilities to change free cytosolic Ca2+ concentrations ([Ca2+]i), adenosine 3',5' cyclic monophosphate (cAMP) levels, and cell proliferation in the osteoblastic cell line, UMR-106-01, and primary cultures of osteoblasts prepared from neonatal rat calvariae. All PGs tested stimulated an increase in [Ca2+]i, which was mainly due to Ca2+ release from intracellular stores. Measurements of the 50% effective concentration for the different PGs show that the potency ranking for PG-evoked [Ca2+]i increase in these cells is F2 alpha greater than D2 much greater than E2 greater than TxB2 greater than E1 greater than I2 much greater than A2. The PGs also increase cAMP levels in osteoblasts. At the highest concentrations tested (10-25 microM), dose-response saturation of cAMP production was observed only by PGE2 and PGE1. The potency rank for PG-stimulated cAMP increase was E2 greater than E1 much greater than A2 greater than I2 greater than F2 alpha greater than D2 greater than TxB2. Measurements of the effect of the PGs on thymidine uptake showed that low concentrations of PGF2 alpha and PGD2 had either no effect or stimulated proliferation of osteoblast-like cells. Relatively low concentration of PGE2, PGE1, and PGA2 inhibited proliferation. The potency ranking for PG mediated inhibition of cell proliferation was identical to that found for PG stimulated cAMP production. We conclude that all the naturally occurring PGs tested can activate the two signal transduction systems in osteoblasts.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556036 TI - Modulation of the ionic permeability of renal cortical brush-border membranes by cAMP. AB - The effects of adenosine 3',5'-cyclic monophosphate (cAMP) on potassium chloride permeability, relative ionic permeabilities, and Na+-dependent glucose transport were examined in rat renal cortical membrane vesicles. Brush-border membrane vesicles were prepared from paired control and cAMP- or forskolin-exposed homogenates by use of a magnesium aggregation technique. These studies demonstrate that exposure to exogenous cAMP or increases in endogenous cAMP significantly increase KCl permeability and ionic chloride permeability relative to that of potassium (PCl/PK) as determined with the fluorescent potential sensitive probe 3,3'-dipropylthiadicarbocyanine iodide [diS-C3-(5)] and 36Cl uptake. Because PNa/PK did not change, PCl/PNa was also significantly increased by cAMP. These changes in ionic permeabilities were associated with a significant stimulation of Na+-dependent glucose transport that was unassociated with either an alteration in the kinetics of glucose transport or delayed dissipation of the inwardly directed Na+ gradient. These findings indicate that the cAMP-induced stimulation of glucose transport resulted from hyperpolarization of the vesicles secondary to the increase in PCl/PNa. These studies suggest that in vivo variations in intracellular cAMP concentration may modulate electrogenic transport processes by altering relative ionic permeabilities and membrane potential in renal proximal tubule cells. PMID- 2556037 TI - Heterogeneity of cAMP effect on endosomal proton transport. AB - The effects of 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP) on proton transport in endocytotic vesicles (i.e., endosomes) obtained from rabbit renal cortex were investigated. Endosomes were prepared from New Zealand White rabbits following the intravenous injection of horseradish peroxidase. Under high (106 mM) chloride conditions, 5 microM 8-BrcAMP stimulated proton transport by 11 and 12% in fractions 1 and 2, but inhibited proton transport by 10, 8, and 14% in fractions 4, 5, and 6. Two hundred and fifty micromolar 8-BrcAMP inhibited proton transport progressively from 20% in fraction 1 to 50% in fraction 6. Under conditions in which chloride entry was minimized, i.e., low (6 mM) external chloride concentration plus voltage clamping, 5 microM 8-BrcAMP consistently and significantly inhibited proton transport by 40-60% in all six fractions. Under the same low chloride conditions, 250 microM 8-BrcAMP inhibited proton transport by 80-90% in all six fractions. These studies indicate that 8-BrcAMP can modify endosomal transport in a heterogeneous manner, that 8-BrcAMP inhibits the H+ ATPase directly, and that this effect is significantly modified by chloride. Last, these studies suggest that the observed heterogeneity in proton transport arises from interaction of 8-BrcAMP and chloride. PMID- 2556038 TI - Basolateral sodium-coupled acid-base transport mechanisms of the rabbit proximal tubule. AB - We studied Na+-coupled acid-base transport at the basolateral membrane of single, isolated, perfused rabbit proximal tubules by monitoring the time course of intracellular pH (pHi). The latter was determined using a microspectrofluorometric apparatus to alternatively excite the pH-sensitive fluorescent dye 2',7'-bis-2-carboxyethyl-5(and -6)-carboxyfluorescein (BCECF) at 440 and 490 nm, while the fluorescence emission, was measured at 530 nm. All experiments were conducted in the nominal absence of HCO-3 S1, S2, and S3 segments from both superficial and juxtamedullary nephrons were examined individually. We found that removing Na+ from both the lumen and bath (i.e., basolateral solution) caused pHi to fall from 7.24 to 6.75 in the superficial S1 segment (SS1), from 7.14 to 6.67 in the SS2, and from 7.09 to 6.69 in the SS3. Similarly, in juxtamedullary nephrons (J), bilateral Na+ removal caused pHi to fall from 7.25 to 6.76 in the JS1, from 7.16 to 6.71 in the JS2, and from 7.10 to 6.75 in the JS3. In all six proximal tubule subtypes, returning Na+ to the bath caused pHi to recover (i.e., increase). 4,4'-Diisothiocyanostilbene-2,2' disulfonic acid (DIDS, 50 microM), an inhibitor of HCO-3 transport systems, blocked this Na+-dependent pHi recovery in all three superficial subtypes and the JS3 but had no effect in either the JS1 or JS2. On the other hand, 50 microM ethylisopropyl amiloride (EIPA), an inhibitor of Na-H exchange, blocked the Na+ dependent pHi recovery in the JS1 and JS2 but had no effect in the JS3 or any of the superficial subtypes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556039 TI - Bradykinin activates protein kinase C in cultured cortical collecting tubular cells. AB - Bradykinin inhibits vasopressin-stimulated water transport in cortical collecting tubular cells. The biochemical mechanism of this effect was explored by means of primary cultures of rabbit cortical collecting tubular cells. Bradykinin was found to produce a rapid release of calcium from intracellular stores, an increase in sn-1,2-diacylglycerol levels, and a fivefold increase in membrane bound protein kinase C activity, consistent with stimulation of phospholipase C and activation of protein kinase C in rabbit cortical collecting tubular cells. In addition, bradykinin produced a dose-dependent 46% inhibition of vasopressin stimulated adenosine 3',5'-cyclic monophosphate (cAMP) formation. Pretreatment with the protein kinase C inhibitors, H-7 and staurosporine, reversed the bradykinin-mediated inhibition of vasopressin-stimulated cAMP accumulation. In contrast, pretreatment with either the phospholipase A2 inhibitor, mepacrine, or pertussis toxin did not prevent the inhibitory effect of bradykinin on vasopressin-stimulated cAMP production, suggesting that the effects are not mediated by prostaglandin E2 or activation of a pertussis-toxin sensitive guanine nucleotide regulatory protein (e.g., Gi). Because bradykinin also inhibits isoproterenol-stimulated cAMP formation but does not inhibit either basal-, forskolin-, or cholera toxin-stimulated cAMP accumulation, the site of this inhibition appears to involve the hormone receptor or coupling of the receptor to the stimulatory guanine nucleotide regulatory subunit (Gs). The results demonstrate that bradykinin stimulates phospholipase C leading to activation of protein kinase C, which then inhibits vasopressin-stimulated cAMP production at the level of the hormone receptor or coupling of the receptor to Gs in cultured cortical collecting tubular cells. PMID- 2556040 TI - Plasma membrane calcium pump and 28-kDa calcium binding protein in cells of rat kidney distal tubules. AB - In an effort to extend our studies on Ca2+ pumps to animal models, we developed a new monoclonal antibody (5F10) prepared against the human erythrocyte Ca2+-Mg2+ adenosinetriphosphatase (ATPase) that recognizes a protein of approximately 140 kDa in rat kidney homogenates. Enzyme-linked immunosorbent assays show that monoclonal antibody 5F10 binds purified Ca2+-Mg2+-ATPase and rat kidney membrane extracts in a concentration-dependent manner. In paraffin-embedded tissue sections, antibody 5F10 binds to an epitope in the basolateral membranes of rat kidney distal convoluted tubule principal cells. The antibody does not bind to intercalated cells. The latter cells were characterized by the presence of large amounts of carbonic anhydrase C. Polyclonal antibodies directed against chick intestinal 28-kDa vitamin D-dependent calcium binding protein (28-kDa CaBP) also bind epitopes in distal convoluted tubule cells, connecting tubules, and portions of collecting duct but not intercalated cells. Western blot and 45Ca blot analysis of renal cytosolic proteins showed that the polyclonal 28-kDa CaBP directed antibody detects a protein which also binds calcium. Western blot analysis with monoclonal antibody 5F10 shows binding to both the authentic purified erythrocyte Ca2+ pump (approximately 138 kDa) and to tryptic fragments of this pump. Antibody JA3, previously used for staining of human kidney tubules, reacts with a different set of tryptic fragments, showing that the two antibodies are directed against different regions or conformational determinants on the pump molecule. We show that Ca2+-Mg2+-ATPase and 28-kDa CaBP are present in the principal cells of the distal convoluted tubule of the rat and are absent in intercalated cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556041 TI - Effects of beta-adrenergic stimulation with isoproterenol on glomerular hemodynamics. AB - To evaluate the contribution of beta 1-2-adrenergic receptor stimulation to the regulation of single-nephron glomerular filtration rate (SNGFR), we examined by micropuncture techniques the effects of systemic and intrarenal infusion of isoproterenol on glomerular hemodynamics in plasma volume-expanded Munich-Wistar rats. Isoproterenol infused systemically was consistently associated with an elevation in glomerular capillary hydrostatic pressure difference (delta P) from 44.2 +/- 1.2 to 50.1 +/- 1.3 mmHg, P less than 0.01, the consequence of a 5.9 mmHg fall in Bowman's space hydrostatic pressure, P less than 0.005. The potentially beneficial effect of increased delta P on SNGFR was overcome by a 40% reduction in the glomerular ultrafiltration coefficient (LpA) from 0.043 +/- 0.003 to 0.026 +/- 0.003 nl.s-1.mmHg-1.g kidney wt-1, P less than 0.005, with a net effect of a modest 13% decline in SNGFR, P less than 0.01. In contrast, the intrarenal infusion of isoproterenol did not modify glomerular hemodynamics. Suppression of angiotensin II activity eliminated the influences of systemic isoproterenol infusion on LpA and delta P, the latter was the consequence of lower efferent arteriolar resistance. The findings suggest that systemic infusion of a beta 1-2-adrenergic agonist results in a decrease in LpA via angiotensin II effects and exerts a vasodilatory action on postglomerular vessels during angiotensin II inhibition. PMID- 2556042 TI - Escherichia coli enterotoxin receptors: localization in opossum kidney, intestine, and testis. AB - The distribution of receptors for Escherichia coli enterotoxin were examined in opossum kidney, intestine, and testis. E. coli enterotoxin stimulated guanosine 3',5'-cyclic monophosphate (cGMP) production in renal cortex, testis, and small intestinal mucosa but had only a small effect in the colon. Atrial natriuretic factor enhanced the cGMP content of renal cortex and small intestine but had no effect on testis or colon. The enterotoxin receptors were observed to be localized in proximal tubules, to epithelial cells of crypts and villi of small intestine, to crypts of colon, and in seminiferous tubules. Both convoluted and straight portions of proximal tubules exhibited specific binding sites for 125I labeled enterotoxin. Glomeruli and distal tubules did not have receptors. Binding of 125I-enterotoxin to brush-border membranes of kidney cortex or intestinal mucosa and to testis membranes was markedly temperature dependent. The binding affinities of these receptors for E. coli enterotoxin were similar (i.e., IC50 approximately equal to 0.4-0.5 nM). Daily administration of 20 micrograms of enterotoxin intramuscularly to opossums increased urine cGMP excretion with no apparent changes in urine volume, Na+, or K+ excretion. Thus receptors for heat stable enterotoxins are localized to proximal tubules of kidney and to enterocytes and seminiferous tubules of intestine and testis, respectively. Apical membranes may be the site of enterotoxin receptors in these epithelia. PMID- 2556043 TI - Inhibition of Jv by ANF in rat proximal straight tubules requires angiotensin. AB - The effects of atrial natriuretic factor (ANF) on fluid absorption (Jv) by isolated perfused proximal straight tubules of rats were investigated. ANF alone (10(-8) M) added to the bath had no significant effect on absorption. In contrast, when tubules were first treated with 1.6 X 10(-10) M angiotensin II, this same concentration of ANF lowered fluid absorption from 0.99 +/- 0.03 to 0.69 +/- 0.02 nl.mm-1.min-1. A lower dose of ANF, 2 X 10(-10) M, reduced fluid absorption in the presence of angiotensin II from 1.13 +/- 0.06 to 0.65 +/- 0.05 nl.mm-1.min-1, an inhibition of 40%. Since guanosine 3',5'-cyclic monophosphate (cGMP) is reportedly part of the second messenger system of ANF, the effects of dibutyryl-cGMP (DBcGMP) on fluid absorption were studied. This membrane-permeant form of cGMP mimicked the effects of ANF, reducing fluid absorption from 1.15 +/- 0.18 to 0.54 +/- 0.08 nl.mm-1.min-1. These studies suggested the following: 1) ANF can regulate fluid absorption in the proximal nephron; 2) this inhibition occurs only in the presence of angiotensin; and 3) cGMP is part of the second messenger system of ANF in the rat proximal straight tubule, as it is in other tissues. PMID- 2556044 TI - Adenosine inhibits ischemia-reperfusion-induced leukocyte adherence and extravasation. AB - Ischemia and reperfusion (I/R) of the small intestine initiates a series of events that result in neutrophil-mediated microvascular injury. Recent reports suggest that adenosine possesses anti-inflammatory properties by virtue of its ability to inhibit neutrophil (PMN) superoxide (O2-.) and hydrogen peroxide (H2O2) production and to interfere with PMN adherence to cultured endothelium. In an attempt to further characterize the anti-inflammatory properties of adenosine in vivo we assessed the influence of exogenous adenosine on 1) I/R-induced PMN mediated microvascular injury in the feline small intestine, 2) feline PMN superoxide production, and 3) I/R-induced PMN adherence to feline mesenteric venular endothelium. We found that intra-arterial administration of adenosine (2 microM) significantly attenuated the I/R-induced increases in intestinal capillary permeability. This protective effect of adenosine could not be explained entirely on its ability to inhibit PMN O2-. (or H2O2) production, since adenosine was effective in inhibiting feline PMN O2-. production by only 20%. Using intravital microscopic techniques in cat mesentery, we found that adenosine did not alter the responses of venular blood flow, shear rate, leukocyte rolling velocity, and leukocyte adherence to I/R when compared with control animals. However, the number of extravasated leukocytes during the ischemic period was significantly reduced by adenosine. Adenosine reduced the number of adherent leukocytes by 25% at 10 and 60 min of reperfusion while leukocyte extravasation was reduced by 65-70% during the same period. Our data indicate that the adenosine-induced suppression of leukocyte extravasation cannot be explained solely by an attenuation in leukocyte adherence to venular endothelium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556046 TI - Adrenal blood flow and secretory relationships during hypoxia in anesthetized dogs. AB - To evaluate whether hypoxia-induced increases in adrenal cortical (CQ) and medullary (MQ) blood flow (radiolabeled microspheres) occur secondary to hypoxia induced secretory activity, pentobarbital-anesthetized ventilated dogs were pretreated with dexamethasone (DEX) to prevent adrenocorticotropic hormone (ACTH) and corticosteroid secretory changes or underwent unilateral adrenal denervation to prevent adrenal catecholamine secretory responses. In nonsurgically stressed dogs, DEX completely prevented increases in ACTH or corticosteroid levels during reduction of arterial oxygen content to 8 vol% but had no effect on hypoxia induced doubling of CQ. In dogs in which adrenal oxygen consumption (VO2) was measured, DEX reduced VO2 by 50% without altering CQ. Unilateral adrenal denervation prevented hypoxia-induced increases in adrenal catecholamine secretion and MQ but had no effect on the CQ response. These results suggest that hypoxia-induced medullary vasodilation is associated with adrenal catecholamine secretory activity but that increases in CQ occur independent of secretory activity and likely represent direct vascular effects of hypoxia. PMID- 2556045 TI - Release of superoxide-dependent relaxing factor(s) from endothelial cells. AB - In the present work, an experimental system was designed to study superoxide anion radical, implicated as the cause of vascular dilatation. To circumvent its direct effect, we employed a two-bath system. When the endothelial cells (EC) were exposed to electrical field stimulation (EFS) or to a hypoxanthine-xanthine oxidase system in bath A plus its physiological buffer solution suffused on a helical strip of cat basilar artery in bath B, the contraction to 5 hydroxytryptamine (5-HT) was depressed to approximately 40-50% of the control value. The reduction was not elicited on EFS in a state of calcium deficiency or in the absence of EC. The depression could be prevented by pretreatment with superoxide dismutase (SOD), but not with an effective dose of catalase, dimethyl sulfoxide (DMSO), mannitol, or indomethacin. The percent depression of contraction was paralleled by an increase in SOD-inhibitable cytochrome c reduction, which was not associated with cyclic guanosine 3',5'-monophosphate formation. These results suggest that superoxide-dependent relaxing factor is released from EC differently than the endothelium-derived relaxing factor mediated by acetylcholine. PMID- 2556047 TI - Blood-induced superoxide anion generation on the cerebral cortex of newborn pigs. AB - Pigs were equipped with closed cranial windows to measure superoxide anion generation in response to blood placed on the cerebral cortex. Superoxide dismutase (SOD)-inhibitable nitro blue tetrazolium (NBT) reduction was measured in the presence of 1) artificial cerebrospinal fluid (CSF), 2) homologous nonheparinized blood (blood), and 3) blood after treatment with indomethacin. SOD inhibitable NBT reduction was increased significantly in the blood group compared with control, whereas piglets pretreated with indomethacin had significantly less SOD-inhibitable NBT reduction. This suggests that superoxide anion is generated by extravascular blood and that this superoxide anion generation can be inhibited by indomethacin. To investigate the cellular origin of superoxide anion generation, SOD-inhibitable NBT reduction was measured during incubation with piglet whole blood or its components in vitro. The SOD-inhibitable NBT reduction of nonheparinized whole blood was 30 +/- 5.7 nmol.ml-1.20 min-1. This decreased to 2.5 +/- 1.3 nmol.ml-1.20 min-1 in the presence of indomethacin. SOD inhibitable NBT reduction of ADP-stimulated platelet-rich plasma was 25.6 +/- 2.9 nmol.400 X 10(6) cells-1.20 min-1 and was decreased significantly (6.8 +/- 3.0 nmol.400 X 10(6) cells-1.20 min-1) in the presence of indomethacin. SOD inhibitable NBT reduction by granulocytes was significant but unchanged by indomethacin. Similarly, SOD-inhibitable NBT reduction by lymphocytes and monocytes was unaffected by indomethacin. Reduction by the red cell fraction was small. These results suggest that substantial quantities of superoxide anion are generated via the platelet cyclooxygenase pathway of arachidonic acid metabolism. PMID- 2556048 TI - Isoproterenol reduces thrombin-induced pulmonary endothelial permeability in vitro. AB - The ability of the beta-adrenergic agonist, isoproterenol, to attenuate the thrombin-induced increase in endothelial permeability was examined by measuring 125I-labeled albumin clearance across endothelial cell monolayers. Bovine pulmonary artery endothelial cells (CCL-209) were grown to confluence on gelatinized, polycarbonate micropore filters and mounted on modified Boyden chambers with Dulbecco's modified Eagle's medium (DMEM) and 0.5% bovine serum albumin. alpha-Thrombin at 0.2 nM to 2 microM produced a dose-related increase (P less than 0.01) in 125I-labeled albumin clearance from the DMEM control value. Light and electron microscopy revealed that the thrombin-induced increase in permeability correlated with changes in cell shape and rearrangement of filamentous actin. Coincubation of 2 microM isoproterenol with 2 microM alpha thrombin reduced (P less than 0.01) the thrombin-induced increase in albumin clearance and the observed morphological changes. This attenuation was not caused by inhibition of thrombin's enzymatically active site, since isoproterenol did not impair thrombin's fibrinogen clotting activity nor its amidolytic cleavage of an artificial substrate (Spectrozyme-TH). Coincubation of 20 microM propranolol, a beta-adrenergic antagonist, with 2 microM isoproterenol and thrombin blocked the permeability-decreasing effect of isoproterenol. Both 2 microM isoproterenol and 2 pM alpha-thrombin alone decreased (P less than 0.01) albumin clearance below the DMEM control value. These results suggest that isoproterenol can reduce the thrombin-induced increase in endothelial permeability in vitro by directly maintaining actin filaments and the shape of endothelial cells. PMID- 2556049 TI - Xanthine oxidase-induced injury to endothelium: role of intracellular iron and hydroxyl radical. AB - The major objective of the present study was to characterize the sequence of events leading to endothelial cytotoxicity induced by oxidants generated extracellularly by xanthine oxidase. 51Cr-labeled monolayers of calf pulmonary artery endothelial cells were exposed to a reaction mixture containing hypoxanthine, xanthine oxidase, and chelated iron (HX/XO) and endothelial cell injury was quantitated as 51Cr release into the media. Catalase, but not mannitol or superoxide dismutase, prevented endothelial cell injury induced by HX/XO, indicating that H2O2 was the mediator of the cytotoxicity. Pretreatment of the cells with free deferoxamine (an iron chelator), but not with deferoxamine bound to dextran (mol wt 40,000), prevented endothelial cell injury induced by HX/XO or H2O2. Of the membrane-permeant hydroxyl radical scavengers dimethylsulfoxide and dimethylthiourea, only dimethylthiourea prevented 1) HX/XO or H2O2-induced endothelial cytotoxicity and 2) deoxyribose degradation by hydroxyl radicals (.OH) generated by an iron-catalyzed reaction on the sugar (site-specific reaction). The concentration of ferritin required to produce significant quantities of .OH was much greater than that present in endothelial cells, and ferritin-catalyzed .OH formation was not affected by deferoxamine, indicating that ferritin-bound iron is most likely not the physiologically active catalyst. We conclude that extracellularly generated H2O2 can enter the cell and interact with nonferritin iron to produce the cytotoxic .OH via a site-specific reaction. PMID- 2556051 TI - Superoxide mediates reperfusion-induced leukocyte-endothelial cell interactions. AB - The objective of this study was to determine whether superoxide mediates the leukocyte-endothelial cell interactions elicited by reperfusion (reoxygenation) of ischemic (hypoxic) tissues. Mesenteric and intestinal blood flows were reduced to 20% of control for 1 h, followed by 1 h of reperfusion. Sixty minutes after reperfusion, red blood cell velocity (Vr), leukocyte rolling velocity (Vw), and the number of adherent leukocytes were measured in mesenteric venules. Then, either human superoxide dismutase (hSOD), hydrogen peroxide-inactivated hSOD, or MoAb IB4 (a monoclonal antibody against the leukocyte adhesion molecule CD18) was injected intravenously. Ten minutes later, repeat measurements were obtained and compared with pretreatment values. hSOD attenuated reperfusion-induced neutrophil adherence and increased Vw/Vr, an index of the fracture stress between leukocytes and endothelium. Peroxide-inactivated hSOD did not alter any parameter. MoAb IB4 attenuated reperfusion-induced adherence but did not alter Vw/Vr. In a correlate study, cultured bovine microvascular endothelium was exposed to 30 min of anoxia, followed by 60 min of reoxygenation. Cat neutrophils were added during reoxygenation. Reoxygenation-induced leukocyte adherence was attenuated by either hSOD or MoAb IB4 but not by inactivated hSOD. Adherence of phorbol 12-myristate 13-acetate-activated cat neutrophils to plastic was unaffected by hSOD or inactive hSOD, yet MoAb IB4 virtually abolished the response. These results indicate that superoxide mediates reperfusion-induced leukocyte adherence and that endothelial cells are required for this superoxide-mediated adherence. PMID- 2556050 TI - Ethacizin blockade of calcium channels: a test of the guarded receptor hypothesis. AB - The effect on calcium channels of the sodium channel antagonist, ethacizin, was studied in isolated frog ventricular cells using the whole cell voltage-clamp methodology. Ethacizin was found to block inward calcium current in a frequency-, voltage-, and concentration-dependent manner. The frequency-dependent blocking properties were modeled by considering the drug interaction with a voltage dependent mixture of calcium channels harboring either an accessible or an inaccessible binding site. With repetitive stimulation, the pulse-to-pulse reduction in peak current is shown to be exponential, with a rate linearly related to the interstimulus interval and the drug concentration. Observed frequency- and concentration-dependent blocks were consistent with the predictions of the model, and mixture-specific rate constants were estimated from these data. The negligible shift in channel inactivation and the reduction of apparent binding and unbinding rates with more polarized membrane potentials imply the active moiety of ethacizin blocks open channels and is trapped within the channel at resting membrane potentials. The binding rate at 0 mV is similar to that observed in studies of interactions of other open channel blocking agents with voltage- and ligand-gated channels. PMID- 2556053 TI - Chemoreceptor stimulation on sympathetic activity: dependence on respiratory phase. AB - Experiments were performed on chloralose-anesthetized, vagotomized, paralyzed, and artificially ventilated cats breathing 100% O2. Peripheral chemoreceptors were stimulated by rapid injections of CO2-saturated NaHCO3 in different phases of the respiratory cycle. Responses of cardiac and renal sympathetic nerves were computed by digital integration. Spontaneous sympathetic activity was consistently modulated by respiration, the modulation being greater for cardiac than for renal nerves. Cardiac nerve responses to peripheral chemoreceptor stimulation depended on the respiratory phase for at least one experimental condition in four of seven animals: the responses were largest during late inspiration and smallest (or absent) during postinspiration and early expiration. Renal nerve responses depended on respiratory phase in only two of eight animals. An average end-tidal CO2 concentration increase from 4.6 +/- 0.8% (SD) to 6.7 +/- 0.9% enhanced the respiratory modulation of spontaneous activity but reduced the responses to peripheral chemoreceptor stimulation. The results indicate that the respiratory modulation of chemoreceptor-induced sympathetic responses was less prominent than the modulation of spontaneous activity. It is hypothesized that the phase dependence of the responses is caused by the spontaneously occurring expiratory diminution of sympathetic activity rather than by an inherent gating of the chemoreceptor reflex. PMID- 2556052 TI - Tedisamil inactivates transient outward K+ current in rat ventricular myocytes. AB - The action of tedisamil, a new bradycardiac agent with antiarrhythmic properties, was investigated in single rat ventricular myocytes using the whole cell voltage clamp technique. Under current clamp conditions, 1-20 microM tedisamil caused marked prolongations of the action potential. Over the same concentration range, in voltage-clamped myocytes, tedisamil suppressed the transient outward current (ito) and enhanced its inactivation in a dose-dependent manner. The half-maximal dose for the effect of tedisamil on ito was approximately 6 microM. Tedisamil had no significant effects on the inwardly rectifying potassium current and calcium current but did suppress the sodium current at concentrations greater than 20 microM. Our findings suggest that tedisamil represents a new type of antiarrhythmic agent that primarily suppresses the transient outward K+ current. PMID- 2556054 TI - Turtles and rats: a biochemical comparison of anoxia-tolerant and anoxia sensitive brains. AB - When temperature differences are taken into account, turtle brains use glucose at one-sixth the rate reported in rat brains. Na+-K+-ATPase activities are 2- to 2.5 fold higher in rat than in turtle brains. Maximal activities of hexokinase and lactate dehydrogenase are similar, whereas citrate synthase activities are two- to threefold higher in rat than turtle brains at the respective biological temperatures. Voltage-dependent Ca2+ channel densities, when compared between the two species, showed no consistent pattern. These data, along with the threefold differences in density of voltage-dependent Na+ channels reported by Lutz et al., are consistent with the idea that lower rates of channel and pump-mediated Na+ and K+ fluxes result in lower rates of aerobic energy metabolism in turtle brains compared with rat brains. PMID- 2556055 TI - Electrophysiological and cable parameters of perfused beetle malpighian tubules. AB - Isolated perfused Malpighian tubules of the desert beetle Onymacris plana (Coleoptera: Tenebrionidae) have been subjected to cable analysis under the following conditions: control, adenosine 3',5'-cyclic monophosphate (cAMP), corpora cardiaca homogenate (CCH), and high ambient K (130 mM). In addition, we investigated possible effects of perfusate composition on proximal transtubular potential (Vo) by reducing K, Na, or Cl or by adding ouabain, furosemide, or dinitrophenol. The effects of cAMP, CCH, and high K on Vo and cable parameters were consistent with increased fluid secretion, i.e., diminished input and core resistances and increased virtual short-circuit current, length constant, and luminal diameter. They differed in that CCH had variable effects on Vo and high K did not reduce transepithelial resistance. In terms of their effects on the parameters of a simple equivalent electrical circuit, the responses to cAMP, CCH, and a high ambient K concentration appear to be mediated by different mechanisms. Alterations in perfusate composition were almost without effect. PMID- 2556056 TI - Association of cytomegalovirus infection with increased morbidity is independent of transfusion. AB - Forty-three consecutive trauma patients with an injury severity score greater than 20 were studied prospectively for evidence of cytomegalovirus (CMV) infection. Twenty-one patients had serologic conversion: 3 with primary CMV infections, 18 with reactivation of CMV infection (CMV group). Twenty-two patients had no serologic conversion (no CMV group). To differentiate the effects of CMV and transfusion, the CMV group and the no CMV group were each divided into high (more than 10 units) and low (less than 10 units) transfusion subgroups. Similar fever peaks, leukocyte counts, lymphocyte counts, and incidence of major bacterial sepsis were recorded for the four subgroups. Several factors were significantly associated with CMV infection independent of transfusion, including increased duration of major bacterial sepsis and number of septic episodes per patient; prolonged duration of anergy; increased duration of intensive care unit and hospital stay; increased duration of ventilatory assistance and rate of tracheostomy; and increased suppressor cells, decreased helper: suppressor ratios, increased functional suppressor cells, and increased natural killer cells. Although mortality was not increased with CMV infection, our data suggest that such infection after trauma may delay recovery from major bacterial infection, often resulting in a major increase in morbidity. PMID- 2556057 TI - Intravenous enalaprilat and autonomic reflexes. The effects of enalaprilat on the cardiovascular responses to postural changes and tracheal intubation. AB - Thirty healthy patients, who were to undergo surgery which required tracheal intubation, were given an intravenous injection of enalaprilat (either 0.5 mg, 1 mg, 2 mg or 4 mg; six patients for each dose) or normal saline 17 minutes before induction of anaesthesia with thiopentone 3-5 mg/kg, and suxamethonium 1.5 mg/kg. Postural manoeuvres were performed 5 minutes before and 6, 11 and 16 minutes after enalaprilat or saline. Complete inhibition of angiotensin converting enzyme occurred with all doses of enalaprilat, which allowed the four different treatment groups to be considered as one large treated group. The mean arterial pressure was almost unchanged during the postural manoeuvres; the heart rate increased, mostly similarly (by approximately 10%) in both groups. Mean arterial pressure in the recumbent position decreased over the 17 minutes before induction in the enalaprilat group, and increased slightly in the control group (treated mean, -5.0%; controls mean, 1.8%; difference, -6.8%; 95% confidence intervals of difference, -2.3 to -11.3%, p less than 0.01). This difference was again seen after induction (treated, -8.0%; controls, 7.7%; confidence intervals of difference, -0.6 to -31%) and for a 5-minute period shortly after tracheal intubation. The increases in mean arterial pressure produced by intubation itself were similar in both groups (treated, + 36%; controls, + 35%; 95% confidence intervals of difference, -16% to + 18%). Changes in heart rate after induction were also similar in both groups. It is concluded that intravenous enalaprilat acted as a hypotensive agent with a sparing effect on autonomic reflexes, both before and after induction of anaesthesia. PMID- 2556058 TI - Postoperative hypotension associated with enalapril. AB - We describe the case of a patient treated for hypertension with the angiotensin converting enzyme inhibitor enalapril, who developed hypotension after recovery from anaesthesia. PMID- 2556059 TI - [Preoperative plasma exchange in treatment of plasma-related coagulation disorders before liver transplantation]. AB - PATIENTS AND METHODS: Seventy-two consecutive patients undergoing orthotopic liver transplantation at the Department of Surgery I, University of Vienna Medical School (OLT nos. 1 to 72), were evaluated. Their mean age was 47 years (range: 18-63 years). The indications for liver transplantation are listed in Table 1. All transplant procedures were performed without using a bypass technique. The intraoperative management and surgical procedure have been described elsewhere [7]. Patients were categorized in two groups, each of which was divided in two subgroups. Group I consisted of 18 patients transplanted before the introduction of preoperative plasma exchange. These were retrospectively allocated to two subgroups on basis of their preoperative prothrombin times (PT): A (n = 9): preoperative PT less than 40%; B (n = 9): preoperative PT greater than 40%. The two subgroups of group 2, which contained 54 patients, were compared on a prospective basis: C (n = 32): preoperative PT above 40%; D (n = 22): PT on admission below 40%, preoperative plasma exchange. Comparison of the two subgroups was based on the following parameters: (1) pre exchange PT (subgroup D); (2) preoperative PT (= PT post-plasma exchange in subgroup D; (3) intraoperative infusion volumes (balanced electrolyte solutions and human albumin to maintain an intravascular colloid osmotic pressure greater than 16 mm Hg); (4) transfusion volumes (whole blood stored for no more than 72 h or packed red cells and fresh plasma, as available; and (5) intraoperative sodium bicarbonate requirements to maintain an arterial pH greater than 7.20. RESULTS: (Table 2) . Prothrombin time (PT): Group 1: Patients in subgroup A had a mean preoperative PT of 34% (range: 15%-40%). This was significantly lower than in subgroup B (74%; 52%-100%; P less than 0.001). Group 2: The pre-exchange mean PT in subgroup D was 27% (12%-39%) vs. 68% in subgroup C (45%-104%), the difference being highly significant (P less than 0.0001). In patients in subgroup D a mean plasma volume of 3638 ml was exchanged by plasmapheresis. This resulted in a significant increase in PT to 55% (Table 3). As a result, the preoperative post exchange PT in subgroup D was slightly but significantly (P less than 0.005) less than in subgroup C. Transfusion volumes: Group 1: Patients in subgroup A needed significantly more blood units than those in subgroup B (55.3 units [19-110] vs. 18.7 [3-33]).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2556060 TI - Intraneural recording of muscle sympathetic activity during epidural anesthesia in humans. AB - The extent and magnitude of sympathetic blockade during epidural anesthesia have previously been assessed only by indirect methods. In this study, direct intraneural recordings of muscle sympathetic activity (MSA) in the peroneal nerve was performed to determine the profundity of blockade of sympathetic fibers to the lower extremities during epidural anesthesia. Lumbar epidural catheters were inserted in nine volunteers. Multiunit postganglionic sympathetic activity was recorded in a muscle fascicle of the peroneal nerve before and after epidural injection of 4 mL of 2% mepivacaine followed by an additional 12-16 mL after 5 min. Apnea (30-60 s) was used to elicit transient sympathetic activation. The upper level of sensory blockade was T-3 to T-10. Muscle sympathetic activity decreased after epidural blockade with no spontaneous or apnea-induced sympathetic bursts observed later than 11 min after injection of the initial test dose. Sympathetic blockade was accompanied by increase in foot skin blood flow as well as loss of skin resistance responses to arousal. Results show that epidural anesthesia with sensory blockade above T-10 to T-11 blocks spontaneous peroneal MSA as well as the marked sympathetic activation induced by apnea. PMID- 2556061 TI - Intraoperative administration of the intravenous angiotensin-converting enzyme inhibitor, enalaprilat, in a patient with congestive heart failure. PMID- 2556062 TI - Are all myocardial infarctions alike? PMID- 2556063 TI - Postoperative myocardial infarction documented by technetium pyrophosphate scan using single-photon emission computed tomography: significance of intraoperative myocardial ischemia and hemodynamic control. AB - The aim of this prospective study was to document postoperative myocardial infarction (PMI) by technetium pyrophosphate scan using single-photon emission computed tomography (TcPPi-SPECT) in 28 patients undergoing elective coronary bypass grafting (CABG). The relationships of intraoperative electrocardiographic myocardial ischemia, hemodynamic responses, and pharmacological requirements to this incidence of PMI were correlated. Radionuclide cardioangiography and TcPPi SPECT were performed 24 h preoperatively and 48 h postoperatively. A standard high-dose fentanyl anesthetic protocol was used. Twenty-five percent of elective CABG patients were complicated with PMI, as documented by TcPPi-SPECT with an infarcted mass of 38.0 +/- 5.5 g. No significant difference in demographic, preoperative right and left ventricular function, number of coronary vessels grafted, or aortic cross-clamp time was observed between the PMI and non-PMI groups. The distribution of patients using preoperative beta-adrenergic blocking drugs or calcium channel blocking drugs was found to have no correlation with the outcome of PMI. As well, no significant differences in hemodynamic changes or pharmacological requirements were observed in the PMI and non-PMI groups during prebypass or postbypass periods, indicating careful intraoperative control of hemodynamic indices did not prevent the outcome of PMI in these patients. However, the incidence of prebypass ischemia was 39.3% and significantly correlated with the outcome of positive TcPPi-SPECT, denoting a 3.9-fold increased risk of developing PMI. Prebypass ischemic changes in leads II and V5 were shown to correlate with increased CPK-MB release (P less than 0.05) and tends to occur more frequently with lateral myocardial infarction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556064 TI - Effect of excitatory amino acid receptor blocker MK-801 on overall, neurologic, and morphologic outcome after prolonged cardiac arrest in dogs. AB - Excitatory amino acids accumulating in the brain during ischemia may cause selective neuronal damage postischemia. This hypothesis was tested in a series of studies using MK-801, an N-methyl-D-aspartate (NMDA) receptor blocker, in a reproducible outcome model of prolonged cardiac arrest in dogs. After normothermic ventricular fibrillation cardiac arrest, the dogs were resuscitated with closed-chest femoral veno-arterial cardiopulmonary bypass. At 4 h they were separated from bypass, ventilation was controlled for 20 h, and intensive care was continued to 96 h. In Study I, ventricular fibrilation cardiac arrest (no flow) was 17 min; starting immediately with reperfusion, MK-801 1200 mg/kg (n = 5) or an equal volume of placebo (n = 5) was infused over 12 h in blinded, randomized fashion. In Study II, the duration of the no-flow period was reduced to 15 min, and MK-801 2400 mg.kg-1 (n = 4) or placebo (n = 4) was infused. In Study III, no-flow lasted for 15 min, and MK-801 2400 mg/kg was started 30 min before ventricular fibrillation (n = 4); comparison was with Study II controls. In all three studies, MK-801 plasma concentrations peaked at greater than 50 ng/ml and were 15-30 ng/ml over 12 h. All 22 dogs of experiments within protocol survived with severe brain damage. MK-801 delayed return of pupillary reactivity, EEG activity, consciousness, and respiration, necessitating longer periods of controlled ventilation. Neurologic deficit scores, overall performance categories, and brain and heart morphologic damage scores at 96 h did not differ between placebo and MK-801 pretreatment or post-treatment groups. These negative outcome results after prolonged cardiac arrest do not negate the hyperexcitability hypothesis of selective vulnerability, but suggest the existance of additional mechanisms of secondary brain damage. PMID- 2556065 TI - Pharmacokinetics and disposition of pipecuronium bromide in dogs with and without ligated renal pedicles. AB - The pharmacokinetics of pipecuronium bromide have been studied in anesthetized beagle dogs with and without ligated renal pedicles. A gas chromatographic assay was used to measure the plasma, urine, bile concentrations, and liver content of pipecuronium, the later of which was obtained 8 h after injection. Following an iv bolus injection of 0.1 mg/kg, pipecuronium disappeared from the plasma exponentially with distribution half-lives of 3.9 +/- 1.1 min and 12.7 +/- 9.5 min (mean +/- SD), and elimination half-lives of 44.8 +/- 2.6 min and 196.7 +/- 102.0 min in animals with and without renal pedicle ligation, respectively. Except for the volume of central compartment, all other pharmacokinetic variables differed significantly between the two experimental groups. The elimination half life was longer (196.7 +/- 102 (SD) vs. 44.8 +/- 2.6 min), plasma clearance slower (5.9 +/- 0.8 ml.kg-1.min-1 vs. 0.9 +/- 0.1 ml.kg-1.min-1) and mean residence time longer (221 +/- 73 vs. 51.1 +/- 1.8 min) in dogs with ligated renal pedicles. Eight hours after injection, the recovery of the parent form of pipecuronium approximated 77% of the administered dose in the urine, 4.5% in the bile, and 3.3% in the liver of normal animals. In animals with ligated renal pedicles 16% of the unchanged pipecuronium was excreted into the bile and 10% of the administered dose was recovered from the liver. Since the total recovery of unaltered pipecuronium approximated 85% of the administered dose in the intact animals, biotransformation seems to play an insignificant role in disposition of this new neuromuscular blocking drug.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556066 TI - [Comparative calibration of dosimetry methods used around industrial irradiation plants in Italy]. AB - Industrial application of ionizing radiation (sterilization of medical supplies, long lasting food preservation, etc.) is a well established technique all over the world. The efficacy of the treatment depends on an accurate dosimetry assuring both the achievement of irradiation purposes at the lowest cost and the safety of consumers. This is why in 1986 the Istituto Superiore di Sanita (ISS) started an intercomparison program among the industrial plants now working in Italy, aimed to optimize the used dosimetric procedures. The electron spin resonance (ESR) alanine-based dosimetry, set up at ISS, was chosen as reference dosimetry. Each plant received 30 dosimeters to be irradiated in prefixed conditions in order to: a) compare the ESR and the routine dosimetry; b) evaluate the homogeneity factor in the irradiated product; c) test the reproducibility of the irradiation technique. Results support the need of standardized dosimetric procedures for an optimization of the radiation treatment. PMID- 2556067 TI - Dermal eccrine cylindroma. AB - Certain dermal appendage tumors have a striking histologic and clinical homology with some tumors arising from the salivary duct system. The dermal eccrine cylindroma is an exemplar. Arising from the eccrine ducts, this neoplasm is only occasionally found outside the head and neck, has a recurrence rate of over 40%, and in its malignant form is a high-grade carcinoma. Its counterpart in salivary glands, the dermal analogue tumor, manifests a histologic similarity, if not identity, also has a relatively high recurrence rate, and can also undergo malignant change. PMID- 2556068 TI - [Heart transplant. Long-term follow-up]. AB - The use of cyclosporin in the immuno-suppressive treatment has resulted in a spectacular improvement of the outcome of cardiac transplantation. But a number of complications, if they have become less severe, have note however totally disappeared. In order to detect them at an early stage, a strict monitoring is necessary. Systematic ambulatory visit are organized in the transplant center. In the interval between visits, a medical follow-up by cardiologist and attending physician is absolutely necessary. PMID- 2556069 TI - Oncogenes-antioncogenes and virus therapy of cancer. AB - Viruses can render services to mankind. 1. Retroviruses pinpoint and transduce cellular oncogenes. 2. Retroviral vectors can introduce antioncogenes (the RB gene) into malignant cells thus rendering the recipient cells nonmalignant. 3. Oncolytic viruses lyse tumor cells. 4. Parvoviruses replicate only in dividing cells and exert lysis and antioncogene effect in tumor cells without affecting resting normal cells. 5. Myxo- and paramyxoviruses (and other viruses) upgrade the immunogenicity of cell surface antigens thus eliciting rejection type host immunity against these cells which is operational against not virus-infected cells of the same type (post-oncolytic antitumor immunity). 6. Viruses or virally infected cells (including tumor cells) induce the production of lymphokines and cytokines (interferons, interleukins and tumor necrosis factor) and activate NK cells and specific immune T cells cytotoxic to virus-infected cells (including tumor cells). 7. Measles virus may activate suppressor cells and both directly (by infecting lymphoma cells) and indirectly (by inducing molecular mediators of suppressor mononuclear cells inhibitory to the growth of neoplastic lymphoid and hematopoietic cells) induce remissions of lympho- and hematopoietic malignancies. 8. Retroviral vectors deliver genes into tumor cells for encoding new surface antigens that render the tumor cells highly antigenic and more vulnerable to rejection type immune reactions of the host. Examples illustrate each statement. Immunotherapy of tumors with active tumor-specific immunization after the induction of suppressor cells by fetal antigens and the elimination of the proliferating suppressor clones by cyclophosphamide will again be proposed. PMID- 2556070 TI - Transformation of primate cells by chemicals and oncogenes: requirements for multiple factors. AB - Although several chemical carcinogenes have been shown to induce malignant transformation in human cells in culture, the molecular events involved in conversion of normal cells to malignant cells remain unknown at present. Normal human cells seem to be resistant to transformation by a single oncogene unless these cells have been immortalized by chemical carcinogens or DNA tumor virus genes. In some human cell systems, malignant phenotype is expressed after activation of protooncogenes probably through the mechanism of point mutations. Our results represent the first induction of KRAS gene rearrangement in cells of patients with familiar polyposis coli (FPC) treated with N-methyl-N'-nitro-N nitrosoguanidine (MNNG) and 12-0-tetradecanoyl-phorbol-13-acetate (TPA). Whether or not KRAS rearrangement is a prerequisite for transformation in these cells is unknown. The contrast between the response of the FPC cells following treatment with these chemicals compared with that of normal fibroblasts suggests that KRAS gene rearrangement in FPC cells may be considered as a genetic marker for the disease. Studies of transformation with chemicals and oncogenes extended to nonhuman primate--marmoset cells have shown that two viral oncogenes, v-src and v sis, converted marmoset cells to altered morphology, anchorage independence and immortality in cell culture. The transformed cells, however, lacked tumorigenicity in allogeneic and xenogeneic hosts. In v-src transformed cells additional changes associated with mutations of transforming virus and chromosome rearrangements were necessary for induction of tumorigenicity in allogeneic marmosets. Transformation of marmoset cells by chemical carcinogens depended on the type of target cells. Skin fibroblasts treated with MNNG acquired transformed morphology, anchorage independence and random chromosome aberrations. Kidney cells treated with the same carcinogen remained untransformed, but attained the same characteristics of transformation when exposed to TPA. Neither skin nor kidney transformed cells showed true tumorigenic potential in nude mice. PMID- 2556071 TI - Molecular and cellular biology of Wilms' tumour. AB - Wilms' tumour is an embryonal kidney tumour which exists in an hereditary and sporadic form. Apart from its obvious importance as a model for renal development and differentiation, the tumour has recently been exploited as an example of the action of tumour suppressor genes (or anti-oncogenes). The latter genes are characterised by a somatic loss of genetic information in tumour development, specifically from the short arm of human chromosome 11 in Wilms' tumour. To further study the developmental aspects of the tumour we have established in vitro cell cultures from tumour tissues, which, unlike the majority of Wilms' tumour cell lines, have been genotyped according to their chromosome 11 gene status and their antigen expression patterns, compared to the original normal kidney and tumour tissues. The cell cultures exist both as primary and secondary cultures, and their limited life span in culture has been extended by transfection of SV40 large T antigen. The mechanism of tumour suppression by the Wilms' locus has been explored by producing cell hybrids between the immortalised kidney cells, and an "indicator cell" (HeLa), whose chromosome 11 genotypes have been monitored in vivo and in vitro by restriction fragment length polymorphisms. Non-random patterns of inheritance of the mutant allele have also been investigated, both in tumour tissue and in syndromes, like the Beckwith-Wiedemann Syndrome, which pre-dispose to development of Wilms' tumour (and other embryonal tumours). It is also apparent that allele-specific methylation occurs in Wilms' tumour tissues, probably resulting in changes of gene expression patterns. Significant elevation of transcription of the N-myc oncogene was detected in the blastemal cells of the most malignant Wilms' tumours, whereas a marked decrease in the expression of HLA class I, at both RNA and protein levels was observed in the same cells. Wilm's tumour provides a clear illustration of the requirement for a combination of dominantly and recessively acting genes, in order to produce a malignant embryonal tumour. PMID- 2556072 TI - The cell biology of human papillomavirus transformed cells. AB - Viruses are becoming increasingly recognized as a major etiological agent in the development of numerous forms of human cancer. Human papillomaviruses (HPVs) have been associated with a number of neoplastic lesions, most notably cervical cancer which is one of the major forms of cancer world wide. Of the over 50 types of identified HPVs, HPV types 16, 18, 31 and 33 are the types most commonly associated with malignant carcinomas. These viruses contain double stranded DNA which code for about eight gene products, some of which are oncogenic when introduces into cultured rodent or human cells. In particular, both the E6 and E7 gene products have different oncogenic capabilities and these genes are selectively retained within the genome of cervical carcinoma derived cells. The E7 gene product has immortalizing capabilities in primary cells and is able to cooperate with an activated ras oncogene to fully transform primary rodent cells. The E7 gene product from HPV type 16 is also capable of complexing in vitro to the anti-oncogene product, Rb. Similar complexes occur with Adenovirus E1A and SV40 large T proteins which may suggest a shared mechanism of transformation used by HPV type 16, Adenovirus and SV40. Transformation studies using primary human cells and nontumorigenic HeLa/fibroblast hybrid cells have also suggested that chromosome 11 may be important in suppressing the HPV transformed phenotype. The transformed phenotype may therefore also involve an impaired intracellular control of persisting HPV oncogenic sequences. Although there exists no solid evidence that a cytotoxic T-lymphocyte reaction is mounted against HPV transformed cells, there is evidence that both NK cells and activated macrophages can preferentially kill HPV transformed cells in vitro. Future studies are required to identify possible targets present on HPV transformed cells which are absent on normal cells. PMID- 2556073 TI - Molecular analysis of the inhibitory effect of phosphorylated ribavirin on the vesicular stomatitis virus in vitro polymerase reaction. AB - The effect of phosphorylated ribavirin on the vesicular stomatitis virus in vitro transcription reaction was examined. Analysis of the kinetics observed when the concentrations of nucleoside triphosphates were varied was performed with vesicular stomatitis virus wild-type standard virions. Double-reciprocal and Eadie-Hofstee plots showed competitive inhibition with all natural nucleoside triphosphates when both ribavirin diphosphate (RDP) and ribavirin triphosphate (RTP) were used. The Km values for ATP obtained for the wild-type polymerase were similar to those reported previously. To further characterize the observed inhibition kinetics, in vitro transcription products synthesized in the presence or absence of RDP and RTP were purified by CsCl centrifugation and were primer extended with oligonucleotides specific for either positive-sense leader or nucleocapsid mRNA transcripts. The ratios of leader to nucleocapsid mRNA were measured from primer-extended in vitro transcription products. It was found that the addition of RDP or RTP did not significantly change the in vitro ratio, suggesting that the polymerase is blocked before it enters the 3' end of the template. PMID- 2556074 TI - Synergistic therapy by acyclovir and A1110U for mice orofacially infected with herpes simplex viruses. AB - Clinical effects of the administration of a combination of acyclovir (ACV) and compound A1110U (a 2-acetylpyridine thiocarbonothiohydrazone inactivator of herpes simplex virus [HSV] ribonucleotide reductase) on the development of herpetic skin lesions were studied in athymic and hairless mice infected intracutaneously with different HSV type 1 (HSV-1) strains. ACV was administered topically (5%) or orally (5 mg/ml), while A1110U was applied topically (3%). In all but one experiment, the effect of combination therapy was greater than that calculated for the sum of the individual drug effects in limiting the development of herpetic skin lesions in mice. In several experiments, combination therapy totally eliminated all signs of infection. This synergistic chemotherapeutic efficacy was evident in infections caused by ACV-susceptible as well as several ACV-resistant HSV-1 strains. These results indicate that this combination therapy may provide a significant improvement in clinical responses over single-agent topical therapy. PMID- 2556075 TI - Use of in vitro topoisomerase II assays for studying quinolone antibacterial agents. AB - Several quinolones and antitumor compounds were tested as inhibitors of purified calf thymus topoisomerase II in unknotting, catenation, radiolabeled DNA cleavage, and quantitative nonradiolabeled cleavage assays. The antitumor agents VP-16 (demethylepipodophyllotoxin ethylio-beta-D-glucoside) and ellipticine demonstrated drug-enhanced topoisomerase II DNA cleavage (the concentration of drug that induced 50% of the maximal DNA cleavage in the test system [CC50]) at levels of less than or equal to 5 micrograms/ml. Nalidixic acid, norfloxacin, and oxolinic acid did not induce significant topoisomerase II DNA cleavage, whereas ciprofloxacin did induce some cleavage above background levels. CP-67,015, a new 6,8-difluoro-7-pyridyl 4-quinolone which possesses potent antibacterial activity, inhibited bacterial DNA gyrase at 0.125 micrograms/ml in a nonradioactive DNA cleavage assay. Unlike other quinolones characterized to date, CP-67,015 was shown to strongly enhance topoisomerase II-induced radiolabeled DNA cleavage with a CC50 of 33 micrograms/ml and demonstrated cleavage in a nonradiolabeled DNA cleavage assay with a CC50 of 73 micrograms/ml. The topoisomerase II-mediated cleavage of DNA by CP-67,015 is consistent with its reported clastogenic effect on DNA in cell culture and its positive mutagenic response in mouse lymphoma cells. In vitro topoisomerase II catalytic and cleavage assays are useful for gaining preliminary information concerning the possible interaction(s) of some quinolones with eucaryotic topoisomerase II which may relate directly to their safety (mutagenicity, clastogenicity, or both) in human and veterinary medicinal usage. PMID- 2556076 TI - Structure-epileptogenicity relationship of quinolones with special reference to their interaction with gamma-aminobutyric acid receptor sites. AB - The relationship between the chemical structure and epileptogenic activity of quinolones was investigated. When the quinolones were administered intravenously to mice concomitantly with oral biphenylacetic acid, a major metabolite of the nonsteroidal antiinflammatory drug fenbufen, enoxacin, norfloxacin, ciprofloxacin, and pipemidic acid, which have an unsubstituted piperazine moiety at the 7 position of their parent nuclei, provoked clonic convulsions and subsequent death at doses of 6.25 mg/kg or more in a dose-dependent manner. AM 1091 and T-3262, which have an unsubstituted aminopyrrolidine moiety at their 7 positions, were less epileptogenic than the compounds listed above were. In contrast, ofloxacin, AT-4140, and nalidixic acid, which have piperazine substituted with methyl group(s) or no piperazine moiety at their 7 positions, never induced convulsions, even at doses of 100 mg/kg. Lomefloxacin, which has a 3-methyl piperazine, however, provoked convulsions at doses of 6.25 mg/kg or more. In the presence of biphenylacetic acid, all the test quinolones except nalidixic acid competitively inhibited [3H]muscimol binding to receptor sites for gamma-aminobutyric acid (GABA) in vitro. Nalidixic acid did not inhibit the binding at all, even at the highest concentration tested, i.e., 10(-4) M. The 50% inhibition doses for [3H]muscimol binding varied within 4 orders of magnitude or more, between 10(-8) to more than 10(-4) M for various compounds, and there was a close correlation between the epileptogenic activities of quinolones and their inhibitory potencies for [3H]muscimol binding to GABA receptor sites. These results indicate that the epileptogenic activity of quinolones possibly relates to the GABA-like structures of substituents at their 7 positions, which act as antagonists of GABA receptors. PMID- 2556077 TI - Fatty acid alterations and polymyxin B binding by lipopolysaccharides from Pseudomonas aeruginosa adapted to polymyxin B resistance. AB - Lipopolysaccharides were extracted from freeze-dried cells of Pseudomonas aeruginosa PAO1 (polymyxin B susceptible), isolate A (polymyxin B resistant), and isolate A-reverted (polymyxin B intermediate resistance) by either the phenol chloroform-petroleum ether or the modified phenol-water method. Isolate A and isolate A-reverted had drastic losses of 2-hydroxydodecanoic acid and significant decreases in 3-hydroxydecanoic acid. Concentrations of amide-linked 3 hydroxydecanoic acid were similar in all three strains. Minor alterations were noted in the composition of 3-deoxy-D-manno-2-octulosonic acid, heptose, phosphate, neutral sugars, and amino compounds. The concentrations of rhamnose in isolate A and of rhamnose and glucose in isolate A-reverted were significantly different from those in PAO1. Trace amounts of mannose and other minor unidentified carbohydrates were detected in all strains. Polymyxin B included in isolate. A growth medium complexed with lipopolysaccharides and remained bound throughout purification. PAO1 lipopolysaccharides bound more polymyxin B than did isolate A lipopolysaccharides. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated minor differences in smooth- and rough-form lipopolysaccharides of the different strains. We propose that loss of hydroxy fatty acids from lipopolysaccharides perturbs outer membrane hydrophobicity and is a contributing factor to polymyxin B adaptive resistance. PMID- 2556078 TI - Treatment of experimental cryptococcal meningitis and disseminated candidiasis with SCH39304. AB - We studied the pharmacokinetics and in vivo antifungal action of SCH39304, a new antifungal azole compound, in rabbits. It crossed the blood-cerebrospinal fluid barrier in the presence or absence of meningeal inflammation, reaching approximately 60% of the simultaneous concentrations in serum. In the treatment of experimental cryptococcal meningitis, SCH39304 was as effective as fluconazole in reducing yeast counts in the subarachnoid space. SCH39304 and fluconazole both were highly effective against candida endophthalmitis, sterilizing the vitreous humor and the choroid and retina. SCH39304 suppressed candida endocarditis and reduced yeast counts in the kidney at all doses tested. SCH39304 was effective in the treatment of experimental cryptococcal meningitis and disseminated candidiasis. Further investigations in humans are warranted. PMID- 2556079 TI - Ex vivo study of serum bactericidal titers and killing rates of daptomycin (LY146032) combined or not combined with amikacin compared with those of vancomycin. AB - Twelve volunteers, in two groups of six, received daptomycin at a dose of 1 or 2 mg/kg. In addition, they received in a randomly allocated order amikacin (500 mg), daptomycin-amikacin, and vancomycin (500 mg). Thirty-five clinical isolates, including Staphylococcus aureus, S. epidermidis, Corynebacterium sp. group JK, and Enterococcus faecalis, were tested in vitro for the measure of the serum bactericidal titers and killing rates. The mean peak concentrations of daptomycin in serum 1 h after the administration of 1 and 2 mg/kg were 11 and 20 micrograms/ml, respectively. At 24 h after the administration of 2 mg/kg, the mean level in serum was 1.9 micrograms/ml, which is higher than the MICs for susceptible pathogens. Daptomycin and amikacin provided identical concentrations in serum whether given alone or in combination. Among the six regimens tested, those including daptomycin provided the highest and the most prolonged serum bactericidal titers against S. aureus, S. epidermidis, and E. faecalis. The killing rates measured by the killing curves were correlated with the concentration/MIC and concentration/MBC ratios of daptomycin for all strains tested. Significant killing occurred once the concentration of daptomycin in the serum 4- to 6-fold the MIC or 1- to 1.2-fold the MBC. The combination of daptomycin and amikacin had no effect on either the serum bactericidal titers or the rates of killing. Only vancomycin provided significant killing of the strains of Corynebacterium sp. group JK. PMID- 2556080 TI - Inhibitory effects of quinolones on DNA gyrase of Escherichia coli and topoisomerase II of fetal calf thymus. AB - The in vitro inhibitory effects of quinolones on the bacterial DNA gyrase of Escherichia coli KL-16 and topoisomerase II of fetal calf thymus were compared. All the quinolones tested required higher concentrations to inhibit the topoisomerase II than to inhibit the DNA gyrase, and no correlation existed among their inhibitory activities against both enzymes. However, there was a large difference among the quinolones in their selectivities between the bacterial enzyme and its eucaryotic counterpart. The selectivity of ofloxacin was highest, and the selectivities of CI-934 and nalidixic acid were lowest. PMID- 2556081 TI - Acute graft-vs-host disease in an immunodeficient newborn possibly due to cytomegalovirus infection. AB - Acute graft-vs-host disease occurring during the early weeks of life has been previously reported as a rare disease entity. We report a case of acute graft-vs host disease in a female infant with an immunodeficiency that was thought to be secondary to intrauterine or neonatal cytomegalovirus infection or, less likely, to a severe combined immunodeficiency. The patient presented with a triad of failure to thrive, diarrhea, and maculopapular and petechial rash. The first clue to diagnosis was the skin biopsy finding of an epidermal lymphocytic infiltrate in association with individual necrotic keratinocytes. The diagnosis was confirmed at autopsy. In the absence of an obvious graft, the disease is believed to have been the result of maternofetal T-cell transfer in utero or at delivery. PMID- 2556082 TI - Hidrotic ectodermal dysplasia with diffuse eccrine syringofibroadenomatosis. PMID- 2556084 TI - Serological assessment of acyclovir treatment of herpes genitalis. AB - Oral acyclovir was given to 60 patients with herpes genitalis--20 experiencing a first attack and 40 with recurrent attacks. All patients were followed up for 1 year. Serial serum samples from the patients as well as from 20 controls were studied to determine the effect of therapy on the immune response to herpes simplex virus (HSV). No toxicity was observed, and very few patients had rather insignificant side effects (e.g., diarrhea). The frequency of recurrence (number of recurrences per year) of genital herpes in acyclovir-treated patients was found significantly lower than in controls. More frequent recurrences were observed in those who had high antibody titer in their early convalescent phase sera than in those without or with a low titer of such antibodies. The antibody titers were reduced in those who received acyclovir as compared with controls. The mean time to seroconversion was longer in the acyclovir-treated group than in controls. Oral acyclovir is thus effective and well tolerated in patients with herpes genitalis. Treatment with acyclovir also diminishes the humoral antibody response to HSV, but it does not prevent recurrence. The effects of acyclovir on the immune response to HSV are discussed. PMID- 2556083 TI - Hepatectomy without abdominal drainage. Results of a prospective study in 61 patients. AB - The increasingly simple postoperative course of major surgery has challenged the routine use of drainage after most abdominal surgical procedures. Therefore a prospective study was designed to determine if abdominal drainage could be safely avoided after liver resection and was evaluated in 61 consecutive patients. There was one postoperative death (1.7%) from variceal bleeding. Four other patients (6.7%) developed an abdominal complication: two right subphrenic hematomas requiring reoperation in one case and two incisional ascitic leaks requiring incisional repair in one patient. There was neither a subphrenic abscess nor bile peritonitis. Postoperative hospitalization was 11.5 +/- 3 days in the entire group and 8.5 +/- 1 days in patients without complications. These results suggest that liver resection can be performed safely without abdominal drainage and that the routine use of drains is unnecessary. PMID- 2556085 TI - Isolation and characterization of oxaloacetate decarboxylase of Salmonella typhimurium, a sodium ion pump. AB - Anaerobic growth of Salmonella typhimurium on citrate is Na+-dependent and requires induction of the necessary enzymes during a 20-40 h lag phase. The citrate fermentation pathway involves citrate lyase and oxaloacetate decarboxylase. The decarboxylase is a membrane-bound, Na+-activated, biotin containing enzyme that functions as a Na+ pump. Oxaloacetate decarboxylase was isolated by affinity chromatography of a Triton X-100 extract of the bacterial membranes on avidin-Sepharose. The enzyme consists of three subunits alpha, beta, gamma, with apparent molecular weights of 63,800, 34,500 and 10,600. The alpha chain contains a covalently attached biotin group and binds to antibodies raised against the alpha-subunit of oxaloacetate decarboxylase from Klebsiella pneumoniae. The Na+ transport function was reconstituted by incorporation of the purified enzyme into proteoliposomes. PMID- 2556086 TI - Immunocytochemical study of small round cell tumors in routinely processed specimens. AB - Fourteen commercially available monoclonal antibodies were used to examine, by the indirect immunoperoxidase technique, 20 formaldehyde solution-fixed, routinely processed bone marrow biopsy and surgical biopsy specimens from a group of small round cell tumors with bone marrow involvement, including 6 neuroblastomas, 4 rhabdomyosarcomas, 3 Ewing's sarcomas, 3 Burkitt's lymphomas, and 4 small-cell carcinomas of the lung. Eight of 14 monoclonal antibodies worked well on the routinely processed sections. Antidesmin (D33) (Dako Corp, Santa Barbara, Calif) and anti-muscle actin (HHF35) showed specific immunostaining in all 4 rhabdomyosarcomas included in this study. Anti-epithelial membrane antigen immunoreactivity was positive in all small-cell carcinomas of the lung. Anti leukocyte common antigen (Dako) showed positive immunostaining in lymphomas and did not immunoreact with nonhematopoietic tumors. However, it was important to be aware that some lymphomas may not have been labeled with anti-leukocyte common antigen. All 6 neuroblastomas showed a positive immunoreactivity with anti chromogranin A (LK2H10) and a weak reaction with antisynaptophysin (SY38). Panels of commercially available monoclonal antibodies, including anti-leukocyte common antigen, anti-epithelial membrane antigen, antidesmin (Dako), anti-muscle actin, and anti-chromogranin A, appear to be most useful in the differential diagnosis and staging of small round cell tumors on routinely processed biopsy specimens. PMID- 2556087 TI - Fatal adenovirus pneumonia in a newborn identified by electron microscopy and in situ hybridization. AB - A male infant born at 25 weeks' gestation died at 2 weeks of age from progressive respiratory insufficiency, metabolic acidosis, and renal failure. Autopsy revealed extensive hemorrhage and necrosis in the lungs, as well as hyaline membrane disease. Alveolar and bronchiolar lining cells contained frequent intranuclear inclusions visible by light microscopy that corresponded to arrays of icosahedral particles suggestive of adenovirus by electron microscopy. Confirmation of overwhelming adenovirus infection was made with in situ DNA hybridization. This case demonstrates the advantage of DNA probe analysis for retrospective diagnosis when no adequate specimen is available for culture or antigen detection. This case is also unusual in that a premature newborn had severe adenovirus infection. PMID- 2556088 TI - Malignant fibrous histiocytoma. Heterogeneous patterns of intermediate filament proteins by immunohistochemistry. AB - Patterns of intermediate filament expression of 10 malignant fibrous histiocytomas (MFHs) were immunohistochemically evaluated using acetone-fixed frozen sections. Seven cases represented the storiform-pleomorphic subtype, 2 were of myxoid type, and 1 was of giant-cell type. All cases had been studied by electron microscopy, and no proof for the diagnoses of liposarcoma, rhabdomyosarcoma, and leiomyosarcoma could be obtained. All tumors showed prominent vimentin immunoreactivity in the tumor cells. Cytokeratin-positive neoplastic cells were found in 2 cases, and in the majority of tumor cells in 1 of these. The 68k neurofilament-positive cells were found in 2 cases. Desmin was not found beyond doubt in the neoplastic cells in any cases, and all cases were negative for glial fibrillary acidic protein. The expression of several types of intermediate filament indicates divergent differentiation properties in MFH and may suggest the heterogeneity of this entity, but more cases should be studied to elaborate any possible consistent patterns of intermediate filament expression in different types of MFH. The expression of multiple types of intermediate filament proteins in MFH can alternatively signify random activation of the corresponding genes in the primitive tumor cells. The complex patterns of intermediate filament proteins in morphologically defined MFHs should be taken into account in the practical immunohistologic analysis of tumors. PMID- 2556089 TI - Oncocytic carcinoma of the male breast. AB - Primary oncocytic carcinoma of the breast was diagnosed in a 76-year-old man. The histologic and electron microscopic features are characteristic of oncocytic neoplasms that occur at other sites, such as salivary glands and endocrine organs. The pathologic features and the presence of estrogen receptors biochemically are distinct from apocrine carcinoma. The patient is alive 7 years after modified radical mastectomy without additional chemotherapy or radiotherapy. PMID- 2556090 TI - Clinical patterns of cytomegalovirus disease after liver transplantation. AB - During a 43-month period, we performed 248 liver transplantations in 211 patients (127 adults and 84 children). Cytomegalovirus (CMV) disease was documented in 73 recipients (34.6%). Risk factors for CMV disease included donor CMV seropositivity, antilymphocyte therapy, and retransplantation. The mean time of occurrence of CMV disease was 38.3 days after transplantation, and the most frequent site of disease was the hepatic allograft. A total of 69 patients were treated with intravenous ganciclovir, with a prompt and lasting response documented in 51 (73.9%). The remaining 18 (26.1%) developed recurrent CMV disease, which was more common after primary CMV exposure. Cytomegalovirus disease was ultimately controlled by ganciclovir in 94.2% of cases. This disease occurs early after transplantation and can be related to well-defined risk factors. Although ganciclovir therapy is effective, preliminary experience with prophylaxis shows promise in reducing the incidence of CMV disease. PMID- 2556091 TI - Leukotriene B4 generation in patients with established pulmonary failure. AB - We investigated the cause of the reduced leukotriene B4 (LTB4) production seen in neutrophils from patients with established adult respiratory distress syndrome compared with control neutrophils. Lymphocytes/monocytes from controls were found to synergistically enhance the amount of LTB4 produced when incubated with neutrophils. This synergistic effect was not seen in cells from patients with adult respiratory distress syndrome. Fatty-acid analysis of neutrophils from patients with adult respiratory distress syndrome and controls showed remarkable similarity in all quantities of fatty acids measured except for arachidonic acid, where there was a 22% reduction in patients' cells compared with controls. Assay of the rate of generation of LTB4 and its degradation product, 20-hydroxy LTB4, revealed that reduced LTB4 generation in patients' neutrophils was not due to increased degradation of LTB4 by hydroxylase enzymes. When the amount of LTB4 being generated per milliliter of whole blood was analyzed in the patients with adult respiratory distress syndrome and compared with controls, it was determined that the potential to generate LTB4 in patients in the intensive care unit was three to five times greater than in controls. PMID- 2556092 TI - Effect of ibudilast on mucociliary transport using frog palate. AB - For the determination of mucociliary transport, we have used a method to measure mucociliary transport force (MCTF), which was defined as the tension developed by the mucus flow and ciliary motility of a frog palate between a force transducer and a glass boat on the palate. 3-isobutyryl-2-isopropylpyrazolo [1,5-a] pyridine (ibudilast) is supposed to increase intracellular cyclic adenosine monophosphate (cAMP) level by the inhibition of cAMP phosphodiesterase and to potentiate the effect of prostacyclin. The effect of ibudilast on MCTF was investigated for 15 min after the drug application. Ibudilast induced a significant, dose-related increase in MCTF at concentrations of higher than 10(-7) M. Therefore, these results indicate that ibudilast, an anti-allergic agent, activates mucociliary transport in respiratory tract. PMID- 2556093 TI - [Development of a system for monitoring the production of leukotriene B4, C4, D4 and E4 in whole blood of pediatric asthmatic patients]. AB - We have developed a new method using a computerized photodiode-array spectrophotometer (CPAS), to characterize the biosynthesis and the inhibition of leukotriene (LT) B4, C4, D4 and E4 in calcium ionophore (Ca-I)-stimulated whole blood. The results obtained were as follows: 1. The use of CPAS enabled us to identify and measure LTs from 2 ml volume of Ca-I stimulated whole blood as well as to check the purity of LTs, without any use of radioimmunoassay or bioassay. 2. LTC4 produced in whole blood was almost completely converted to LTE4 after 80 min incubation, and at this time both the production of LTE4 and LTB4 reached a plateau and remained constant thereafter. 3. Azelastine added in vitro caused a dose-dependent inhibition of Ca-I stimulated LTB4 and LTE4 production with an IC50 of 10 microM. PMID- 2556094 TI - [Polyamines stimulate the phosphorylation of phosphatidylinositol in rat mast cell granules]. AB - Rat mast cell granules were obtained by homogenization of highly purified rat mast cells and isolated in a Percoll gradient. Diphosphoinositide (DPI) synthesis in rat mast cell granules was assayed by measuring the incorporation of 32P from [gamma 32] ATP into DPI in the absence of exogenous phosphatidylinositol (PI). Lipids were isolated with methanol/chloroform/HCl and were separated by thin layer chromatography on oxalic acid impregnated silica gel plates. DPI areas were identified by staining with iodine, scraped and measured for 32P radioactivity. The addition of polyamines, spermine and spermidine to the granules caused an increase in DPI synthesis, which can be catalyzed by PI kinase. This effect of polyamines on DPI synthesis in the rat mast cell granules was dose-dependent and maximal effects were observed at 1 mM spermine and 10 mM spermidine respectively. When the effect of 1 mM spermine on 32P incorporation into DPI in rat mast cell granules was investigated serially, 32P incorporation into DPI in rat mast cell granules incubated with spermine for 15 min was enhanced significantly (p less than 0.05) compared with that in the granules in the absence of spermine. PMID- 2556095 TI - Vascular endothelial cell migration in vitro roles of cyclic nucleotides, calcium ion and cytoskeletal system. AB - According to the response to injury hypothesis, endothelial migration and repair may play an important role in the initiation and progression of atherosclerosis. In this study, we examined the regulatory mechanisms of endothelial cell migration in vitro, using cultured endothelial cells from fetal bovine aortas. Dibutyryl cyclic AMP, 8-bromo cyclic GMP, and theophylline (each at concentrations of 10(-4) to 10(-3) M) inhibited the migration of endothelial cells. Migration was not significantly affected by the Ca2+ channel blockers diltiazem (10(-6) to 10(-4) M) and nicardipine (10(-6) to 10(-5) M) or by La3+ (10(-4) to 10(-3) M), an inorganic Ca2+-antagonist, TMB-8 (10(-6) to 5 x 10(-5) M), an intracellular Ca2+ blocker, or the calmodulin inhibitors W-7 (10(-6) to 5 x 10(-5) M) and trifluoperazine (10(-7) to 10(-5) M). At the extracellular Ca2+ concentrations of less than 0.2 mEq/l, the migration was inhibited significantly. In addition, migration was markedly suppressed by colchicine (10(-8) to 10(-5) M), an inhibitor of tubulin polymerization, and by cytochalasin B (10(-7) to 10( 5) M), an inhibitor of actin polymerization. These results suggest that cyclic nucleotides, such as cyclic AMP and GMP, may regulate the migration of vascular endothelial cells. Although a low concentration of extracellular Ca2+ is essential to their migration, participation of the intracellular Ca2+-calmodulin system was not evident in this study. It appears that the cytoskeletal system, including microtubules and microfilaments, is involved in the mechanisms of migration. PMID- 2556097 TI - Perioperative nerve lesions. AB - The scenario is all too familiar. A patient has a surgical procedure. Anesthesia is uneventful and the procedure goes well. Nothing untoward is observed in the recovery room. Later that evening the patient complains of numbness, weakness, or pain, and a neurological deficit is found. A neurologist is called, examines the patient, and it is concluded that a nerve lesion has occurred, because of stretch, ischemia, compression, or laceration. A subsequent electromyogram and nerve conduction study confirm the clinical impression, but offer little in the way of explanation. Over the subsequent months, the patient makes a slow recovery but there has been a prolonged period of pain and disability. Liability issues loom. This scenario could reflect a number of different nerve lesions. This review illustrates the different modes of pathogenesis that may occur and the variable nature of the neurological deficits. We grouped these lesions according to the clinical setting in which they occur. PMID- 2556096 TI - Glucose uptake by gliomas after treatment. A positron emission tomographic study. AB - Positron emission tomographic scanning with fludeoxyglucose F 18 (18F fluorodeoxyglucose) was used to study acute changes in gliomas after chemotherapy. In six experimental subjects, scans were obtained before and at days 1, 7, and 30 after treatment. Five control patients with gliomas who did not undergo chemotherapy had two scans, 1 month apart. Ratios were calculated between peak tumor regional cerebral metabolic rate for glucose and contralateral white matter. The percent change in ratios relative to each patient's baseline scan was calculated. Ratios in three stable controls remained unchanged over the study interval; in two controls it increased 155% and 36% and both died of tumor progression. In experimental subjects, ratios increased 20% to 100% 24 hours after chemotherapy and then decreased until at 28 days they varied between 22% above and 35% below baseline. The increased fludeoxyglucose F 18 uptake at 24 hours could be from uncoupling oxidative phosphorylation or shunting glucose to ribose phosphates for salvage nucleoside synthesis. PMID- 2556098 TI - Effects of eicosapentaenoic and docosahexaenoic acids on apoprotein B mRNA and secretion of very low density lipoprotein in HepG2 cells. AB - Oleic acid (18:1n-9, OA), docosahexaenoic acid (22:6n-3, DHA), or eicosapentaenoic acid (20:5n-3, EPA) was added to HepG2 cells at a concentration of 1 mM in a 5:1 or 2:1 molar complex with bovine serum albumin (BSA), and this was incubated for 3 hours. The incorporation of 3H-glycerol into cellular and medium triglyceride (TG), and the mass of TG were measured. The effects of these fatty acids on the secretion of very low density lipoprotein (VLDL) apolipoprotein B (apo B) were estimated from the incorporation of 3H-leucine into the medium apo B in comparison to cells incubated with fatty acid-poor albumin. The secretion of human albumin by the cells was also estimated by immunochemical precipitation of the labeled albumin. In addition, the intracellular levels of apo B messenger ribonucleic acid (mRNA) were measured by the dot-blot hybridization technique. Relative to control cells incubated with BSA, OA (complexed to BSA at a 5:1 molar ratio) stimulated TG synthesis and secretion sevenfold. Compared to OA, EPA was 24% less effective for both processes, whereas DHA inhibited only the secretion of TG (-43%). The secretion of VLDL apo B was not affected by OA, but was decreased 31% by EPA and 54% by DHA. When the molar ratio of fatty acid complexed to albumin was changed to 2:1, similar results were obtained with respect to TG production. The levels of apo B mRNA relative to actin mRNA were not significantly altered by any of the fatty acids.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556099 TI - Unlikely association between clinically apparent herpesvirus infection and coronary incidence at older ages. The Framingham Heart Study. AB - Experimental studies in chickens have shown a relationship of a herpesvirus to atherosclerosis. The hypothesis of an association in humans was tested by using data on the history of cold sores and other manifestations of herpes infections reported by 658 male and 919 female participants (ages 58 to 89) in the Framingham Heart Study from 1977 to 1979 and on the prevalence and subsequent 6 year incidence of coronary heart disease (CHD). Approximately 40% of the men and 52% of the women reported a history of ever having "fever blisters or cold sores." Overall, there was no association between a history of such oropharyngeal manifestations and prevalent CHD. Only in the subgroup of women with recurrent infections was there a suggestion of a possible relationship (relative risk = 1.5, 95% confidence interval 1.0 to 2.1). Among members of the cohort without CHD at baseline there was no association between the history of cold sores, chicken pox, shingles, or infectious mononucleosis and 6-year CHD incidence. However, a possible interaction among women with recurrent herpes, lower levels of serum cholesterol, and incidence of angina pectoris without myocardial infarction was suggested in post hoc analyses. These data from the Framingham cohort do not support the notion that any self-reported clinically manifest herpesvirus infection has a strong etiological role in older persons, but they do raise issues to be addressed in any further research. PMID- 2556101 TI - Serological reactions in sheep and cattle experimentally infected with three Australian isolates of bluetongue virus. AB - Serums from 103 sheep and 24 cattle experimentally infected with one of 3 serotypes of bluetongue virus isolated in Australia were tested for antibody to bluetongue virus in the serum neutralisation test and the agar gel diffusion precipitin test. Antibody to bluetongue virus was first detected by these tests 8 to 10 days after intravenous infection in 4 sheep that were bled daily for serum analysis. The agar gel diffusion test failed to detect antibody in 28% (29/103) of sheep which had seroconverted in the serum neutralisation test. A further 7% (7/103) of sheep serums were negative in both tests 14 to 22 d after infection. Both tests detected antibody to bluetongue virus in all cattle serums by 10 days after detection of viraemia. In comparison with the intravenous route of infection, extended prepatent periods for the commencement of viraemia resulting from intradermal, subcutaneous and intrauterine routes of infection in the cattle caused corresponding delays in the detection of antibody. For example, one cow that was infected by intrauterine inoculation did not become viraemic until 22 d after inoculation and antibody was not detected until 32 d after inoculation. PMID- 2556100 TI - Brain stem encephalitis due to herpes simplex virus. AB - A 6 year old child is described with infection due to herpes simplex virus type 1 causing brain stem encephalitis. The diagnosis was established by enzyme immunosorbent assays of the cerebrospinal fluid and serum which demonstrated antibody responses to herpes simplex virus. Recovery occurred and the importance of early use of acyclovir in achieving a good outcome is emphasized. PMID- 2556102 TI - Pathogenicity for sheep of recent Australian bluetongue virus isolates. PMID- 2556103 TI - Differential influences of corticotropin-releasing factor on memory retention of aversive learning and appetitive learning in rats. AB - The effects of intraamygdala injections of corticotropin-releasing factor (CRF) on memory retention of aversive and appetitive learnings were examined in the present study. Other than confirming the results of a previous study that a moderate dose of CRF (0.1 microgram) injected into the amygdala facilitated retention performance of an inhibitory avoidance task (electric shock punishment) 24 h after training, we have further found that this effect lasted for a week. However, CRF produced a differential dose-response effect upon memory of an appetitive learning paradigm (sexual reinforcement). CRF at 0.01 microgram enhanced retention at both 24-h and 1-week intervals, while CRF at 0.1 microgram impaired retention in the same task at 1 week only. These results confirm that the amygdala is important in memory processing and suggest that the mechanisms involved in the action of CRF on memory retention of aversive and appetitive learnings are different. The possible differences are also discussed. PMID- 2556104 TI - The relationship between the amygdala and bed nucleus of the stria terminalis in the cat: an evoked potential and single cell study. AB - The effects of amygdala stimulation on excitability of cells in the bed nucleus of the stria terminalis (BNST) were investigated in the cat. The predominant effect of stimulation was to excite cells localized in the lateral BNST. Cells responded either with single spikes to a stimulus pulse or in short bursts. Spontaneous firing of cells after a pulse to the amygdala was observed to both increase and decrease over a 4-s interval. Increases in firing rate, however, were the predominant response. Cells in more anterior locations in the BNST responded with latencies shorter than those of cells in more posterior locations, reflecting either differences in conduction time of excitation from the amygdala or differences in transmitters mediating the excitatory effects. Associated with increases in cell firing was a compound field potential with an initial negative component and a later positive component. These components may be generated by different cell types within the BNST. The negative component likely represents a field EPSP. Effective sites of amygdala stimulation were restricted to the posterior basal amygdala, and effects observed in the BNST were restricted to the lateral BNST. These data correspond well with anatomical studies showing a monosynaptic projection of basal amygdala to lateral BNST in the cat. This study suggests that this projection is predominantly excitatory. PMID- 2556105 TI - Intraseptal administration of muscimol produces dose-dependent memory impairments in the rat. AB - The present study examined the effects of intraseptal administration of the GABAergic agonist muscimol on performance of a radial-arm maze (RAM) task. Male Long-Evans rats were trained to perform a RAM task in which a 1-h delay was imposed between the sample and the test session. In this task rats have access to four out of eight maze arms during a predelay session. Following a 1-h delay, rats are returned to the maze and allowed to freely choose among all eight arms. Arms not blocked during the predelay session are baited, and entry into an arm chosen during the predelay session or a repeated entry into a postdelay chosen arm constitutes an error. Following acquisition, animals were implanted with a single cannula aimed at the medial septum. A within-subjects design was utilized to examine the effects of intraseptal administration of muscimol (0.0, 0.75, 1.5 or 3.0 nmol) on performance in this task. All drugs or artificial cerebrospinal fluid were administered immediately following the predelay session. Muscimol, a GABA-A agonist, produced a dose-dependent impairment in maze performance as evidenced by fewer correct choices in the first four postdelay choices and an increase in the number of errors. Intraseptal administration of muscimol did not significantly alter latency per choice on the RAM task nor did it affect locomotor activity levels. Muscimol-induced impairments were also observed when a 4-h delay was imposed between the fourth and the fifth maze selection, suggesting that the behavioral deficit represents an inability to store or retain spatial working memories rather than a general performance deficit. These data indicated that pharmacological manipulation of GABA-A receptors within the medial septum modifies working memory processes. The potential interaction of GABAergic and cholinergic mechanisms in the modulation of working memory processes is discussed. PMID- 2556107 TI - The action of defined oxygen-centred free radicals on human low-density lipoprotein. AB - The effects of defined oxygen-centred free radicals on human low-density lipoprotein (LDL) structure and receptor affinity are discussed in relation to the mechanisms of cell-mediated oxidative modification of LDL. Both hydroxyl (OH.) and hydroperoxyl (HO2.) radicals caused depletion of endogenous alpha tocopherol and formation of hydroperoxides. Superoxide (O2-.) radicals produced only very limited oxidation, but could potentiate oxidation stimulated by the addition of Cu2+. All these radicals enhanced the net negative charge of intact LDL and induced fragmentation of apolipoprotein B-100 (apo B). OH. also caused cross-linking of apo B. Radical attack decreased the affinity of LDL for the fibroblast apo B/E receptor, but did not enhance its endocytosis by mouse macrophages. PMID- 2556106 TI - Calmodulin-binding proteins as calpain substrates. PMID- 2556108 TI - Characterization of agonist-stimulated incorporation of myo-[3H]inositol into inositol phospholipids and [3H]inositol phosphate formation in tracheal smooth muscle. AB - The effects of the muscarinic agonist carbachol, histamine and bradykinin on incorporation of [3H]inositol into the phosphoinositides and the formation of [3H]InsPs were examined in bovine tracheal smooth-muscle (BTSM) slices labelled with [3H]inositol. These agonists result in substantial and dose-related increases in the incorporation of [3H]inositol into the phospholipids. Carbachol and histamine stimulated the incorporation of [3H]inositol into the phospholipids to the same degree, despite histamine being only 35% as effective as carbachol on [3H]InsP accumulation. Histamine and carbachol, at maximal concentrations, were non-additive with respect to both the stimulated incorporation of [3H]inositol and [3H]InsP formation. For carbachol this effect on incorporation was found to occur to a similar extent in PtdInsP and PtdInsP2 as well as PtdIns. The initial effect of carbachol on [3H]inositol incorporation was rapid (maximal by 10 min); however, with prolonged stimulation large secondary declines in PtdInsP and PtdInsP2 labelling were observed, with depletion of the much larger PtdIns pool only evident in the presence of Li+. Lowering buffer [Ca2+] increased the incorporation of [3H]inositol under basal conditions, but did not attenuate the subsequent agonist-stimulated incorporation effect. The large changes in specific radioactivity of the phosphoinositides, and consequently the [3H]InsP products, after carbachol stimulation resulted in the apparent failure of atropine to reverse the [3H]InsP response completely. Labelling muscle slices with [3H]inositol in the presence of carbachol or labelling for longer periods (greater than 6 h) prevented subsequent carbachol-stimulated effects on incorporation without significantly altering the dose-response relationship for carbachol-stimulated [3H]InsP formation and resulted in steady-state labelling conditions confirmed by the ability of atropine to reverse fully the [3H]InsP response to carbachol. This study demonstrates the profound effects of a number of agonists on [3H]inositol incorporation into the phospho- and polyphosphoinositides in BTSM with important consequent changes in the specific radioactivity of these lipids and the resulting [3H]InsP products. In addition, a selective depletion of PtdInsP and PtdInsP2 over PtdIns has been demonstrated with prolonged muscarinic-receptor stimulation. PMID- 2556109 TI - Phosphoinositide hydrolysis in mitogen-stimulated human peripheral-blood T lymphocytes. AB - Both phytohaemagglutinin and antibodies to the CD3 molecule induced proliferation and phosphoinositide hydrolysis in human peripheral-blood T lymphocytes, but the magnitude of the inositol phosphate response was small and the rate of accumulation slow [significant increases in Ins(1,4,5)P3 were observed only after 10 min]. Hence this response differs from the well-characterized Ins(1,4,5)P3 responses of many other systems. This slow response, its abrogation in Ca2+ depleted medium, the slow and maintained increase in Ca2+ as measured by Quin-2, and the ability of the Ca2+ ionophore A23187 to stimulate Ins(1,4,5)P3 accumulation all suggest that the increase in Ins(1,4,5)P3 occurs, at least in part, as a result of receptor-mediated Ca2+ influx in mitogen-stimulated T lymphocytes. PMID- 2556110 TI - Characterization of phospholipids accumulated in pulmonary-surfactant compartments of rats intratracheally exposed to silica. AB - Phospholipids in the lung fractions, i.e. alveolar free cells, extracellular pulmonary surfactant, intracellular pulmonary surfactant (lamellar bodies) and microsomal fractions, of rats were examined 28 days after intratracheal injection of silica (40 mg/kg). Significant accumulations of phospholipids were observed in the extracellular- and intracellular-surfactant fractions of rats exposed to silica. The prominent phospholipid accumulated was phosphatidylcholine (PC), consisting mainly of the dipalmitoyl species. However, a compositional change in acidic phospholipids of surfactant fractions was produced by the silica treatment. The percentage of phosphatidylglycerol (PG) was significantly decreased; in contrast, that of phosphatidylinositol (PI) was increased. Thus the ratio PG/PI in the surfactant fractions was markedly decreased in response to silica. This compositional change in both acidic phospholipids occurred even in the early stages, i.e. before appreciable accumulations of alveolar phospholipids were noticed. The molecular-species profiles of both acidic phospholipids in the surfactant fractions were distinctly different from each other. The dipalmitoyl species accounted for more than 30% of PG and less than 6% of PI, respectively. These species patterns of PG and PI were similar in control and silica-treated rats. These findings suggest two possibilities that (1) PG and PI destined for pulmonary surfactant are synthesized from each specific CDP-diacylglycerol (DG) pool having different molecular species in the lung, or (2) individual enzymes responsible for synthesis of surfactant PG and PI have substrate specificities for molecular species of CDP-DG, thereby producing PG and PI having different molecular species in surfactant compartments. PMID- 2556111 TI - delta-Aminolaevulinate synthase in human HepG2 hepatoma cells. Repression by haemin and induction by chemicals. AB - delta-Aminolaevulinate (ALA) synthase, the rate-limiting enzyme in haem biosynthesis in the normal liver, was examined in human HepG2 hepatoma cells. Haemin, up to 100 microM, had no effect on ALA synthase activity in vitro; it did, however, exhibit a dose-dependent inhibitory action when added to cells growing in culture (half-maximal inhibition at 1 microM). The half-life of ALA synthase activity after haemin treatment was 2 h, which was similar to that found after treatment with cycloheximide. Cells treated with actinomycin D showed a longer half-life of the enzyme activity, i.e. 4 h, compared with haemin or cycloheximide treatment. Treatment of cells with succinylacetone markedly inhibited the activity of ALA dehydratase and 59Fe incorporation into haem, but in increased ALA synthase activity. Both the haemin-induced repression and the succinylacetone-mediated de-repression of ALA synthase activity were reversible within 4 h after replacing the medium with fresh medium without the chemical. In addition to succinylacetone, dimethyl sulphoxide and 3-methylcholanthrene induced the enzyme. Induction of ALA synthase by these chemicals was also suppressed by treatment of cells with haemin. These findings indicate that the level of ALA synthase in HepG2 cells is maintained by both synthesis and degradation of the enzyme, and that the synthesis of the enzyme is regulated by the concentration of regulatory free haem in the cell. PMID- 2556112 TI - Subcellular localization and hormone sensitivity of adipocyte cyclic AMP phosphodiesterase. AB - Treatment of intact adipocytes with either or both insulin and adrenaline stimulated membrane cyclic AMP phosphodiesterase activity only in the endoplasmic reticulum subfraction. The cyclic GMP-inhibited cyclic AMP phosphodiesterase activity was also found in this fraction. Quantitative Western blotting using a specific polyclonal antibody, raised against the homogeneous 'dense-vesicle' cyclic AMP phosphodiesterase from rat liver, identified a single 63 kDa species which was localized in the adipocyte endoplasmic reticulum fraction. The ability of adrenaline to stimulate adipocyte membrane cyclic AMP phosphodiesterase was shown to be mediated via beta-adrenoceptors and not alpha 1-adrenoceptors. Membrane cyclic AMP phosphodiesterase was stimulated by glucagon but not by vasopressin, A23187 or 12-O-tetradecanoylphorbol 13-acetate (TPA). Treatment of adipocytes with either chloroquine or dansyl cadaverine failed to affect the ability of insulin to stimulate cyclic AMP phosphodiesterase activity. Treatment of an isolated adipocyte endoplasmic reticulum membrane fraction with purified protein kinase A increased its cyclic AMP phosphodiesterase activity some 2-fold. When this fraction was treated with purified protein kinase A and [32P]ATP, label was incorporated into a 63 kDa protein which was specifically immunoprecipitated with the antiserum against the liver 'dense-vesicle' cyclic AMP phosphodiesterase. PMID- 2556113 TI - Affinity purification of cellular retinoic acid-binding protein on 14-carboxy-13 cis-retinamide-sepharose 4B. AB - A biologically active bifunctional retinoid, ethyl 14-carboxyretinoate, has been synthesized and shown to bind cellular retinoic acid (RA)-binding protein (CRABP) via its free carboxy group. We describe herein the synthesis of 14-carboxy-13-cis retinamide-Sepharose 4B, which is an affinity matrix bearing an all-trans-RA moiety, and thus was used to purify and characterize CRABP from chick-embryo skin. An amide bond was first formed between the free carboxy group of the retinoid and a primary amino group of aminohexyl-Sepharose 4B, by reaction with carbodi-imide, and the ester group of the resin-bound retinoid was then hydrolysed in an alkaline medium. Polyacrylamide-gel electrophoresis and f.p.l.c. Superose column-chromatographic analysis demonstrated that the affinity-purified CRABP (Mr 15,000) was close to electrophoretic homogeneity (greater than 90%) and specifically interacts with RA. By using affinity gel chromatography, conversion of holo-CRABP into apo-CRABP by treatment with p-hydroxymercuribenzoate and a possible involvement of a thiol group in RA binding to CRABP were established. This affinity procedure provides several advantages: (i) 14-carboxy-13-cis retinamide-Sepharose exhibited high efficiency and selectivity for RA-binding protein (i.e. retinol- or fatty-acid-binding proteins did not bind); (ii) the presence of the amide linkage between the ligand and the matrix makes this affinity resin relatively stable to cytosolic enzymes; and (iii) other RA-binding proteins, e.g. nuclear receptor(s), may be purified. PMID- 2556114 TI - Platelet-derived growth factor stimulates synthesis of 1,2-diacylglycerol from monoacylglycerol in Balb/c 3T3 cells. AB - 1,2-Diacylglycerol (1,2,-DAG) plays an important role in the protein kinase C mediated signal-transduction system. Several reports have shown that 1,2-DAG is generated through various pathways other than classical phospholipid hydrolysis. We observed a rapid incorporation of [3H]myristate into 1,2-DAG in platelet derived-growth-factor (PDGF)-treated Balb/c 3T3 cells. [14C]Palmitate was similarly incorporated into 1,2-DAG. The effect of PDGF, which was inhibited by cycloheximide, became maximal after 60 min treatment with PDGF, and disappeared 300 min after removal of PDGF. Treatment with triacylglycerol lipase revealed that labelled saturated fatty acid was incorporated into the sn-1 position. PDGF barely stimulated incorporation of [3H]glycerol or [14C]glucose into 1,2-DAG. Incorporation of [3H]myristate into 1,2-DAG preceded that into triacyglycerol and phospholipids. These results suggest that synthesis of 1,2-DAG from monoacylglycerol is enhanced in PDGF-treated cells. However, there is no significant difference in the activity of monoacylglycerol acyltransferase measured in vitro in quiescent and PDGF-treated cells. The reason for this discrepancy is discussed. PMID- 2556116 TI - Cholesterol and bile acid synthesis in Hep G2 cells. Metabolic effects of 26- and 7 alpha-hydroxycholesterol. AB - 1. Using a human hepatoma (Hep G2) cell line that continually synthesizes 3 beta hydroxy-5-cholenoic acid, lithocholic acid, chenodeoxycholic acid and cholic acid we have determined the metabolism and biological effects of 26-hydroxycholesterol and 7 alpha-hydroxycholesterol. 2. Addition of 26-hydroxycholesterol to the medium (6 microM) downregulated cholesterol and chenodeoxycholic acid synthesis. 3. The predominant metabolite of 26-hydroxycholesterol was 3 beta-hydroxy-5 cholenoic acid. 4. Cholesterol synthesis was not affected by the addition of 7 alpha-hydroxycholesterol (6 and 12 microM). The predominant metabolite of 7 alpha hydroxycholesterol was chenodeoxycholic acid. 5. In Hep G2 cells 7 alpha hydroxylation of 26-hydroxycholesterol is not well expressed. PMID- 2556117 TI - Poliovirus translation: a paradigm for a novel initiation mechanism. AB - All eukaryotic cellular mRNAs, and most viral mRNAs, are blocked at their 5' ends with a cap structure (m7GpppX, where X is any nucleotide). Poliovirus, along with a small number of other animal and plant viral mRNAs, does not contain a 5' cap structure. Since the cap structure functions to facilitate ribosome binding to mRNA, translation of polio-virus must proceed by a cap-independent mechanism. Consistent with this, recent studies have shown that ribosomes can bind to an internal region within the long 5' noncoding sequence of poliovirus RNA. Possible mechanisms for cap-independent translation are discussed. Cap-independent translation of poliovirus RNA is of major importance to the mechanism of shut-off of host protein synthesis after infection. Moreover, it is likely to play a role in determining poliovirus neurovirulence and attenuation. PMID- 2556115 TI - Ca2+-induced accumulation of pyrophosphate in mitochondria during acetate metabolism. AB - The mechanism of pyrophosphate (PPi) accumulation in rat liver during acetate metabolism was investigated. Perfusion of the liver with acetate in the presence of noradrenaline and glucagon induced marked accumulation of PPi (2 mumol/g of liver, 200 times that of control). In contrast, perfusion with glutamine, which generates PPi only in the cytosol, caused little accumulation of PPi, even in the presence of the two hormones. The site of PPi accumulation was shown to be the mitochondria by the finding that isolated mitochondria from the liver perfused with acetate and the hormones contained 50 nmol of PPi/mg of protein. The addition of an uncoupler to mitochondria with accumulated PPi caused gradual decrease in their PPi content, with concomitant release of a stoichiometric amount of Ca2+. Similar accumulation of PPi was observed when isolated mitochondria were incubated with acetate and Ca2+. These results show that an increase in cytosolic Ca2+ caused by the co-administration of the two hormones induced uptake of the ion into mitochondria, and that PPi accumulated in mitochondria only when it was generated in the organelles with an elevated concentration of Ca2+. High mitochondrial concentrations of Ca2+ are considered to inhibit inorganic pyrophosphatase through the formation of a stable complex, CaPPi-. Mitochondria with accumulated PPi had normal respiratory activities, and their adenine nucleotide concentrations were increased 2-fold rather than being decreased, the increases also being considered to be caused by their high concentration of Ca2+. PMID- 2556118 TI - Localization of c-ERB A proteins in rat liver using monoclonal antibodies. AB - Monoclonal antibodies were raised against the nuclear thyroid hormone receptors encoded by c-ERB A genes and against a purified nuclear receptor fraction. These antibodies recognize the c-ERB A protein in nuclear extracts from rat liver and are able to compete with thyroid hormone in Scatchard analyses. In sections of rat liver they react with all the hepatocyte nuclei as well as with the cells of the hepatic bile ducts. Comparison with another putative T3 receptor antibody, described previously, showed that distinct 57 kD proteins with a different cellular distribution were recognized. PMID- 2556119 TI - Defective sterol C5-6 desaturation and azole resistance: a new hypothesis for the mode of action of azole antifungals. AB - Two azole resistant isolates of Saccharomyces cerevisiae carried mutations allelic to erg 3 and were blocked to differing degrees at the C5-6 desaturation step of ergosterol biosynthesis. When treated with the sterol 14 alpha demethylation inhibitor fluconazole the wild-type sensitive strain accumulated lanosterol and 14 alpha-methyl-erogosta-8,24(28)-dien-3 beta, 6 alpha-diol (14 methyl-3,6 diol). The stringent desaturase mutant, A2, accumulated 14 alpha methyl-8,24(28)-dien-3 beta-ol (14-methyl fecosterol) and lanosterol as the major sterol components when treated with fluconazole. Resistant isolate A3 accumulated 14-methyl-3,6-diol, 14-methyl fecosterol, and lanosterol and was only partially blocked at sterol C5-6 desaturation. We conclude that functional sterol C5-6 desaturase is required for the synthesis of 14-methyl-3,6-diol under conditions of azole inhibition. We present a new hypothesis for the mode of action of azole antifungals based on the inability of 14-methyl-3,6-diol to support growth, and suggest that growth can occur through utilisation of 14-methyl fecosterol, produced by a combination of azole inhibition and defective sterol C5-6 desaturation. PMID- 2556120 TI - Characterization of a Mr 25,000 basic fibroblast growth factor form in adult, regenerating, and fetal rat liver. AB - A heparin-binding Mr 25,000 immunoreactive bFGF-like protein (ir-bFGF) is recognized in adult rat liver extract by affinity-purified polyclonal anti-human placental bFGF antibodies. Hepatic levels of this protein increase 4-fold in regenerating rat liver during the first 48 h after partial hepatectomy. Also, they appear to be higher in embryonic than in newborn or in adult rat liver. Mr 25,000 ir-bFGF from regenerating rat liver, partially purified by heparin affinity chromatography, induces plasminogen activator activity and cell proliferation in transformed fetal bovine aortic endothelial GM 7373 cells and competes with Mr 18,000 [125I]bFGF for the binding to high affinity bFGF receptors. The data indicate the presence in rat liver of a high molecular weight form of bFGF whose expression is modulated during embryonic development and liver regeneration. PMID- 2556121 TI - Ischaemia causes externalization of endothelin-1 binding sites in rat cardiac membranes. AB - Specific, high affinity binding sites for iodinated endothelin-1 ([125I]-ET-1) were identified in crude plasma and light membrane fractions harvested from aerobically perfused and ischaemic rat hearts, to determine whether the ischaemia induced increase in binding site density (Bmax) involves externalization of the sites. In crude plasma membranes Bmax increased after 60 min ischaemia, from 113.5 +/- 2.15 to 180.6 +/- 4.67 fmol/mg protein (p less than 0.01). In the light membranes, the Bmax fell, from 94.7 +/- 8.70 to 63.80 +/- 6.26 fmol/mg protein (p less than 0.05). Hill coefficients and selectivity of both membrane fractions were unchanged. These results are interpreted as meaning that ischaemia causes externalization of cardiac [125I]-ET-1 binding sites. PMID- 2556122 TI - Adrenocorticotropic hormone-releasing activities of interleukins in a homologous in vivo system. AB - We compared adrenocorticotropin-releasing activities of several interleukins in a homologous or heterologous in vivo system. Intravenous injection of rat interleukin-1 alpha significantly increased plasma adrenocorticotropin in conscious, freely-moving rats 30 min after the injection, and the effect was 10 times greater than that of human interleukin-1 alpha. Rat interleukin-2 affected plasma adrenocorticotropin in a much slower manner and increased its levels significantly 120 min after the injection. Human interleukin-2 had no effect on plasma adrenocorticotropin. Thus, species difference in the experimental system should be considered to assess the physiological significance of cytokines in the neuroendocrine system. PMID- 2556123 TI - Use of 3,3,5,5-tetramethyl-1-pyrroline-1-oxide spin trap for the continuous flow ESR monitoring of hydroxyl radical generation in the ischemic and reperfused myocardium. AB - A new approach for in vivo spin trapping and quantitation of oxygen-derived free radicals has been developed using a continuous flow high speed ESR detection system. Spin adducts of OH. were detected as 1:1:1:1:1:1 sextets (aN=15.2 G, aH=16.8 G, g=2.0055) in the isolated rat heart when perfused with 3,3,5,5 tetramethyl-1-pyrroline-1-oxide (40 mM) during a 10-min control pretreatment (14 ml/min) followed by 50 min of low-flow ischemia (1 ml/min), 30 min of global ischemia and subsequent reperfusion at 14 ml/min. The ESR signals appeared within 15-20 min of low-flow ischemia and grew moderately during the remaining 30 min at a rate of 2-6 nmoles of spin adduct released per minute. Post-ischemic reperfusion was characterized by a burst of spin adduct formation at 30 s-1 min, corresponding to 51.8 nmoles of spin adduct released between 30 s and 1 min. PMID- 2556124 TI - Binding of bovine, ovine, porcine, canine, and rat plasminogen to rat hepatocytes and rat C6 glioma cells in vitro. AB - Plasminogens were purified by affinity chromatography from bovine, ovine, porcine, canine, and rat plasma. The binding of each plasminogen to rat hepatocytes in primary culture and to rat C6 glioma cells was studied by radiodisplacement experiments. All of the plasminogens inhibited human 125I [Glu1]plasminogen type 2 binding to specific cell surface receptors. The IC50 values were similar. These studies suggest conservation of the receptor recognition site in plasminogens across species lines. PMID- 2556125 TI - Linear alpha-human atrial natriuretic peptide analogs display receptor binding activity and inhibit alpha-hANP-induced cGMP accumulation. AB - We have synthesized a series of [Cys(R)7,23]alpha-hANP analogs, in which the two Cys residues were modified with various alkyl groups(R); i.e., R=Acm, Pe, Qe, Cam, Me, Ae, Bzl, Cm, Ocam and sulfo. The Acm-, Cam-, and Me-analogs exhibited binding activity as potent as alpha-hANP in rat vascular smooth muscle cells (VSMC). Binding activity of the analogs decreased progressively as the bulkiness of the R group increased. None of the analogs caused accumulation of cGMP in VSMC and vasorelaxant activity in rat aorta. Acm-, Cam- and Me-analogs substantially antagonized alpha-hANP-induced cGMP accumulation, but did not antagonize vasorelaxation induced by alpha-hANP in vitro. PMID- 2556126 TI - Basic fibroblast-like growth factor is present in the conditioned medium of simian sarcoma virus transformed NRK cells. AB - The conditioned medium of Simian sarcoma virus (SSV)-transformed NRK cells contains at least two activities that down regulate the epidermal growth factor receptor. To identify these activities, we analyzed the medium for the presence of factors both related to and distinct from the v-sis oncogene product. Fractionation of the conditioned medium from SSV-transformed NRK cells by chromatography on heparin-Sepharose yielded two active fractions capable of inhibiting EGF binding. The first component, which eluted at 0.8 M NaCl, is able to induce autophosphorylation of the platelet-derived growth factor (PDGF) receptor, is a mitogen for Swiss 3T3 cells and corresponds to the PDGF B chain product of the v-sis oncogene. The second component requires 2 M NaCl for elution, is mitogenic for Swiss 3T3 cells and inhibits high affinity EGF binding through a protein kinase C-independent pathway, all properties of basic FGF. These results suggest that the conditioned medium of v-sis-transformed cells contains at least two factors that can act in an autocrine capacity, one derived from v-sis and one corresponding to basic FGF. PMID- 2556127 TI - Spectroscopic evidence for a copper-nitrosyl intermediate in nitrite reduction by blue copper-containing nitrite reductase. AB - The reactions of nitrogen monoxide (NO) with the blue copper-containing nitrite reductases from Alcaligenes sp. NCIB 11015 and Achromobacter cycloclastes IAM 1013 were investigated spectroscopically. The electron paramagnetic resonance (EPR) signals of the blue coppers vanished in the presence of NO at 77 K, being fully restored by the removal of NO. The additions of NO to the enzyme solutions resulted in the substantial bleaching of the visible absorption bands at room temperature. The reactions were also completely reversible. These results suggest the formation of a cuprous nitrosyl complex (Cu+-NO+), which is likely the intermediate in the enzymatic nitrite reduction. PMID- 2556128 TI - Purification and DNA-binding properties of human papillomavirus type 16 E6 protein expressed in Escherichia coli. AB - Unfused human papillomavirus type 16 (HPV 16) E6 protein was expressed in Escherichia coli using a lambda PL promoter system. The protein was isolated from the cells as inclusion bodies, extracted by 6 M guanidine-HCl, and purified by chromatography. The purified protein had high affinity to DNA and was demonstrated for the first time to bind to a specific sequence within the long control region of HPV 16. PMID- 2556129 TI - Fast effects of aldosterone on electrolytes in human lymphocytes are mediated by the sodium-proton-exchanger of the cell membrane. AB - An in vitro effect of aldosterone on intracellular sodium, potassium and calcium concentrations and cell volume was shown in human mononuclear leukocytes (HML). In the present paper the effect of aldosterone on propionate-induced swelling of HML and, thereby, the Na+/H+-exchanger of the cell membrane was investigated. Aldosterone in physiological concentrations stimulated the cell swelling from 3 min on significantly. This effect had to be attributed to an increased activity of the Na+/H+-exchanger since amiloride blocked it almost completely. The inhibitor of protein synthesis actinomycin D prevented this action, cortisol had an intermediate effect. These data are the first to identify the Na+/H+-exchanger as a primary target of mineralocorticoid action in HML. PMID- 2556130 TI - Neutrophil activation by surface bound IgG is via a pertussis toxin insensitive G protein. AB - Pre-treatment of neutrophils with either pertussis or cholera toxins does not inhibit neutrophil activation by surface bound IgG. In contrast, pretreatment with the phorbol ester, phorbol myristate acetate, results in a dose dependent inhibition of degranulation by surface bound IgG. This inhibition is similar to that seen with soluble ligands where it is thought to be due to interference with the interaction of an activated guanine nucleotide binding protein with phospholipase C (J. Biol. Chem.,262,6121,1987). More directly, GTP binding and GTPase activity are enhanced when human neutrophil membranes are incubated in wells containing surface bound IgG. Neither of these G protein functions were inhibited when membranes were prepared in the presence of pertussis toxin, suggesting that neutrophil activation by surface bound IgG proceeds by a mechanism that involves a pertussis toxin insensitive G protein. PMID- 2556131 TI - Changes in myocardial mitochondrial electron transport activity in rats administered with acetylcholinesterase inhibitor. AB - This study was designed to elucidate harmful effects of acetylcholine on myocardial mitochondrial electron transport activity. Rats were cervically dislocated 3 h and 6 h after oral administration of pyridostigmine, an acetylcholinesterase inhibitor. The myocardial mitochondrial electron-transport activity (NADH-cytochrome c reductase, succinate-cytochrome c reductase and cytochrome c oxidase), and myocardial acetylcholine and norepinephrine concentrations were measured. Activities of cytochrome c oxidase were significantly decreased in the pyridostigmine-3h and the pyridostigmine-6h groups compared with untreated rats. Activity of NADH-cytochrome c reductase was significantly decreased 6 h after administration. No significant changes were observed in those of succinate-cytochrome c reductase among all groups. Pyridostigmine increased significantly myocardial acetylcholine concentration, however, no significant changes of myocardial norepinephrine concentrations were observed among all groups. It is indicated that these mitochondrial injuries might be dependent on an increase in acetylcholine level and independent of norepinephrine. PMID- 2556132 TI - cAMP activates Na+/H+ antiporter in murine macrophages. AB - The role of cAMP in activating the Na+/H+ antiporter in murine macrophage (M phi) system was investigated. Incubation of PU5-1.8 macrophage tumour cells, peritoneal M phi and bone marrow derived macrophages (BMDM phi s) with dibutyryl cAMP (db-cAMP) or cholera toxin (CT) led to an increase in intracellular pH (pHi). The magnitudes of these responses differed markedly in the three cell types, BMDM phi s being the most sensitive, PU5-1.8 cells the least so. These cells also differed in their responses to inhibitors of Na+/H+ exchange. In PU5 1.8 cells, the db-cAMP- or CT-triggered intracellular alkalinization was abolished by amiloride treatment which, however, was ineffective in BMDM phi s. The chemotactic peptide, N-formyl-methionyl-leucyl-phenylalanine (FMLP), also caused a significant increase in cytoplasmic pH. However, its action was apparently not mediated by cAMP. The significance of these observations is discussed. PMID- 2556133 TI - Electron transfer facilitated by superoxide dismutase: a model for membrane redox systems? AB - Membranes, which are an amalgam of proteins and lipids, effect electron transfer through largely unknown mechanisms. Using albumin with bound fatty acids as a model, we have investigated the possible role of these two membrane constituents in electron transfer. In the presence of albumin: fatty acid, there is substantial enhancement of the reduction of ferricytochrome C by ferrous iron. To assess the possible role of free superoxide in cytochrome C reduction, we added mammalian copper/zinc containing superoxide dismutase (Cu/Zn SOD), which catalyzes the transfer of electrons between superoxide anion radicals, forming oxygen and hydrogen peroxide. Surprisingly, in the presence of either albumin or fatty acid free albumin, Cu/Zn SOD actually accelerates electron transfer from ferrous iron to ferricytochrome C. By contrast, neither inactive Cu/Zn SOD nor active manganese SOD facilitates the ferrous iron-dependent reduction of cytochrome C. These results suggest that, in some circumstances, Cu/Zn SOD may transfer electrons to alternative acceptors and that such transfer depends upon the unique reduction/oxidation reaction mechanism of Cu/Zn SOD. If so, this ubiquitous enzyme could be involved in regulating cellular electron transfer reactions as well as acting as a superoxide 'detoxify-ing' agent. PMID- 2556134 TI - Receptors for insulin and insulin-like growth factor-I in the human adrenal gland. AB - Human adrenal glands contain high-affinity receptors for insulin and insulin-like growth factor I (IGF-I). Comparative studies with rat, hamster and human adrenal membranes confirmed that IGF-I receptors are most abundant in rat and hamster adrenals, whereas insulin and IGF-I receptors are present in equivalent numbers in human adrenal glands. Covalent crosslinking studies revealed that the human adrenal gland IGF-I receptor binding subunit migrated on dodecyl sulfate polyacrylamide gels with Mr = 135,000, which is identical to the migration of IGF I receptor binding subunits isolated from other tissues. Autoradiography of frozen human adrenal slices incubated with [125I]insulin showed prominent, displaceable binding of this radioligand to the zona reticularis, zona glomerulosa, vasculature and medulla; in contrast, [125I]IGF-I binding to human adrenal tissue was most prominent in the zona reticularis and negligible in the medullary region. PMID- 2556135 TI - Monoclonal antibodies to the human insulin receptor mimic a spectrum of biological effects in transfected 3T3/HIR fibroblasts without activating receptor kinase. AB - The effects of four monoclonal antibodies to the alpha subunit of the human insulin receptor were studied in transfected mouse 3T3 fibroblasts expressing human insulin receptors (3T3/HIR). Three antibodies, MA-5, MA-20, and MA-51, mimicked insulin stimulation of the uptake of both 2-deoxy-D-glucose and alpha aminoisobutyrio acid, and S6 kinase activity. Antibody MA-5 also mimicked insulin stimulation of [3H]thymidine incorporation and cell growth. Although these antibodies mimicked insulin stimulation of biological effects, they failed to significantly activate insulin receptor tyrosine kinase activity. These studies suggest, therefore, that the insulin receptor can signal a variety of cellular functions without stimulation of receptor kinase activity. PMID- 2556136 TI - Substitution of an extracellular cysteine in the beta 2-adrenergic receptor enhances agonist-promoted phosphorylation and receptor desensitization. AB - We constructed and expressed in a permanent cell line a beta 2-adrenergic receptor with a valine substitution for cysteine 184 of the second putative extracellular loop. The mutant receptor was partially uncoupled from adenylyl cyclase with impaired ability to form the high affinity agonist-receptor-G protein complex, yet displayed more rapid and extensive agonist-induced desensitization. The enhanced desensitization was accompanied by increased agonist promoted, but not cAMP promoted, receptor phosphorylation in intact cells. Thus, not only is impaired desensitization associated with decreased phosphorylation, as we have shown with several mutant beta 2-adrenergic receptors recently, but enhanced desensitization is accompanied by increased agonist promoted receptor phosphorylation. In the case of this cysteine mutant, this may be due to the greater accessibility of the uncoupled receptor for phosphorylation by the beta-adrenergic receptor kinase. PMID- 2556137 TI - Proteolytic degradation routes for turkey beta 1-adrenoceptor probed with antipeptide antibodies against the N-terminal sequence of the receptor. AB - Anti-peptide antibodies, raised against the N-terminal sequence (amino acids 2 10) of the turkey beta 1-adrenoceptor [Yarden et al., Proc. Natl. Acad. Sci. USA (1986) 83, 6795-6799] recognized the 50 kDa- but not the 40 kDa-form of the receptor, thus confirming the previous assumption that the N-terminus of the 50 kDa form is lost during its conversion to the 40 kDa-form [Jur beta, R., Hekman, M. & Helmreich, E.J.M. (1985) Biochemistry 24, 3349-3354]. By in situ proteolysis small amounts of receptor fragments were formed, which could be recognized by the N-terminus specific antibody. Therefore, although the production of the stable 40 kDa receptor species by proteolytic removal of a portion of the N-terminal appears to be the predominant route, there exists an additional pathway of degradation which must involve the initial cleavage of the carboxyl terminal. PMID- 2556138 TI - Generation of alloxan free radicals in chemical and biological systems: implication in the diabetogenic action of alloxan. AB - Electron spin resonance (ESR) studies that on reaction with NADPH, alloxan was reduced forming labile anion radicals giving a 7-line signal with g = 2.005. These radicals were also produced on incubation of alloxan with rat liver subcellular fractions and their production was greatly enhanced by NADPH. Alloxan effectively scavenged superoxide anion generated by a xanthine-xanthine oxidase (XOD) system in association with its reduction to these anion radicals. These radicals were also formed during incubation of alloxan with rat pancreatic beta cells. These results suggest that the cytotoxicity of alloxan is related to the formation of alloxan anion radicals. PMID- 2556140 TI - Long-term phorbol ester treatment dissociates phospholipase D activation from phosphoinositide hydrolysis and prostacyclin synthesis in endothelial cells stimulated with bradykinin. AB - Bovine pulmonary artery endothelial cells (BPAEC) were prelabeled with [3H]choline or [3H]myristic acid to selectively label endogenous phosphatidylcholine. BPAEC were stimulated with ATP and bradykinin (BK), and phospholipase D (PLD) activation was detected as a 4-fold increase in [3H]choline in cells prelabeled with [3H]choline or as a 2- to 3-fold increase in [3H]phosphatidylethanol in cells prelabeled with [3H]myristic acid and stimulated in the presence of ethanol. Pretreatment of BPAEC with 0.1 microM phorbol 12 myristate 13-acetate (PMA) for 22 hr completely inhibited agonist-induced PLD activation, whereas prostacyclin synthesis and [3H]phosphoinositide ([3H]PIns) hydrolysis were enhanced in pretreated cells. Long-term PMA treatment thus dissociates agonist-induced PLD activation from [3H]PIns hydrolysis, and agonist induced prostacyclin synthesis is not dependent upon PLD activation. PMID- 2556139 TI - Depolarization potentiates endothelin-induced effects on cytosolic calcium in bovine adrenal chromaffin cells. AB - The effects of endothelin on intracellular calcium concentrations ([Ca2+i]) in primary cultures of bovine adrenal chromaffin cells (BAM) were measured using Fura 2. Endothelin had minimal effects on [Ca2+i] over a broad dose range (1 nM to 1 microM). However, in conjunction with K+ depolarization there was a synergistic increase in [Ca2+i]. This effect was dependent on extracellular calcium as was the response to KCl alone. A partial synergistic effect was evident with endothelin and nicotinic stimulation. The effects of endothelin and angiotensin II on [Ca2+i] are only additive. Blockade of voltage sensitive calcium channels failed to alter the synergistic effects. Our results indicate that endothelin influences BAM calcium mobilization through sites regulated by membrane depolarization but differing from traditional voltage sensitive calcium channels. PMID- 2556141 TI - Sodium transport defect of ouabain-resistant renal Na,K-ATPase. AB - The murine renal Na,K-ATPase is resistant to cardiac glycosides. It is not yet known however whether altered active transport is associated with the drug resistance. To investigate this problem Na,K-ATPases were purified from the outer medulla of both rat and rabbit kidneys and reconstituted identically into liposomes. The Na-stimulation of the Na,K-ATPase activity before reconstitution and of the Na-transport after reconstitution was measured. A Na-defect inherent in the ouabain-resistant rat Na,K-ATPase was discovered indicating a link between the cardiac glycoside sensitivity and the Na-transport. PMID- 2556142 TI - Neurotensin receptors on circular smooth muscle of canine small intestine: role of disulfide bridges. AB - We have studied the effect of sulfhydryl agents on the binding of 125I-tyr3 neurotensin to the purified plasma membranes from circular smooth muscle and on the in vitro response of circular muscle strips of canine small intestine to neurotensin. Dithiothreitol (DTT) enhanced the binding by about 80%. Cysteine (a reductant) also enhanced the binding while cystine (an oxidant) reduced the binding to the similar extent. DTT stimulated the tissue in the organ bath and abolished the stimulatory response to low concentrations of neurotensin. The stimulatory response to acetylcholine was not altered by DTT. The implications of the role of disulfide bridges in the neurotensin response is discussed. PMID- 2556143 TI - Monokine regulation of glucose transporter mRNA in L6 myotubes. AB - Endotoxin-induced macrophage secretory proteins (monokines) have been shown to stimulate hexose uptake in L6 myotubes (1). In those studies a doubling of the Vmax for hexose uptake was observed which correlated with elevated numbers of glucose transporters (GT) in both plasma and microsomal membranes. To determine if these changes in transporter populations were due to increased GT mRNA, we performed Northern blot analysis using L6 cell RNA and a cDNA to the HepG2 glucose transporter. The L6 myotubes contained a single 2.8 kb species of GT mRNA that increased 2.5-fold after an 8h exposure to the monokine preparation. beta Actin mRNA levels were unaltered by the treatment, indicating specificity of monokine action. Glucose transporter mRNA content appeared to reach a maximum 8 h after exposure to the monokine. Over the next 16 h the levels of this mRNA gradually decreased, approaching control levels. Data obtained from nuclear transcription run-on assays suggest that increased levels of CT mRNA are due to an increased rate of gene transcription. A second transporter, the insulin sensitive glucose transporter, was also observed to be expressed in the L6 cells. Monokine treatment resulted in a 60% suppression of the mRNA coding for this protein. PMID- 2556144 TI - Heat shock induction of heme oxygenase mRNA in human Hep 3B hepatoma cells. AB - Heat shock treatment of human Hep 3B hepatoma cells led to the induction of mRNA for microsomal heme oxygenase. The maximum induction of heme oxygenase mRNA (5--- 7-fold) was observed with treatment of cells at 43.5 degrees C, for 60 min. The heat-mediated induction of heme oxygenase mRNA was blocked by simultaneous treatment of cells with actinomycin D or cycloheximide. In contrast to Hep 3B cells, cells of another human hepatoma line, Hep G2, showed little induction of heme oxygenase mRNA by heat treatment. These findings suggest that heat shock treatment induces heme oxygenase mRNA in certain human hepatoma cells, but not in others. PMID- 2556145 TI - Primary structure of the S. cerevisiae gene encoding uridine monophosphokinase. AB - In the yeast Saccharomyces cerevisiae, the biosynthesis of both pyrimidine nucleoside triphosphates UTP and CTP is dependent on the activity of the uridine monophosphokinase step. We have determined the sequence of the uridine monophosphokinase gene. The coding region is 615 base pairs long and encodes 205 amino acids (22,500 daltons). The 5' terminus is comprised of a 17 amino acid long hydrophobic leader sequence which is not present in genes encoding adenylate kinases. The coding region shows a strong degree of homology with the cytosolic adenylate kinases of vertebrates, and a lesser degree of homology with yeast and E. coli adenylate kinases. PMID- 2556146 TI - Changes of N-methyl-D-aspartate activated channels of cerebellar granule cells with days in culture. AB - N-Methyl-D-Aspartate (NMDA) activated channels were studied in enzymatically dissociated cerebellar granule cells primary cultures. Measurements of single channel currents were made on different days in culture. Changes in the electrophysiological behavior of NMDA-activated channels, which were dependent on the time in culture, were found. The variations of single channel maximum conductance during the developement of the cells in culture were detected. Three different characteristic periods could be distinguished: the first period (1-3 days) in which the conductance assumed a value of 15.5 pS; the second one (5-8 days) characterized by a value of 35.7 pS and the last one (9-11 days) in which the conductance reached values of 46.8 pS. Moreover mean open time of NMDA activated channels was less than 1 msec during the first two days in culture and stabilized at 3 to 6 msec around the fifth day. PMID- 2556148 TI - Low-affinity interaction of fibrinogen carboxy-gamma terminus with human monocytes induces an oxidative burst and modulates effector functions. AB - The interaction of highly purified, monomeric fibrinogen (Fg) with human monocytes (MO) was investigated. In contrast to commercial Fg, no high-affinity binding of monomeric Fg to MO or mononuclear cells could be demonstrated. MO preincubated with Fg in the presence or absence of Ca++ elicited an oxidative burst when triggered with anti-Fg antibodies. Divalency of the antibody and specificity were required, but an intact Fc portion was not. Surface-adsorbed monomeric Fg also promoted an oxidative burst. Evidence is presented that Fg-MO interaction is mediated by the carboxy-gamma terminus of Fg. MO treated with monomeric Fg or exposed to Fg-coated surfaces show a reduced oxidative burst upon triggering with unrelated stimuli. Thus, MO function may be modulated upon interaction with surface-adsorbed Fg or with fibrin. PMID- 2556147 TI - Interleukin 1 beta inhibition of TRH-stimulated prolactin secretion and phosphoinositides metabolism. AB - The effect of interleukin 1 beta on prolactin secretion and on phosphoinositide turnover in anterior pituitary cells was evaluated. Interleukin 1 beta significantly inhibited TRH-stimulated prolactin secretion assessed by the reverse hemolytic plaque assay. In particular, the cytokine reduced the percentage of plaque forming cells, the plaque mean area, the large plaques percentage. TRH-stimulated inositol phosphate production was also significantly inhibited by interleukin 1 beta. This study shows that interleukin 1 beta reduces TRH-induced prolactin secretion through a direct action on pituitary cell, and attenuates the TRH-stimulated phosphoinositide breakdown. This latter effect may suggest that the reduced lactotropes sensitivity to TRH action may be partially due to interleukin 1 beta inhibition of phosphatidylinositol breakdown. PMID- 2556149 TI - [Conformation-activity relations of dermorphin and its pentapeptide analogs]. AB - Low-energy peptide backbone structures of dermorphin (DM), amide of its N terminal pentapeptide (DM 1-5) and DM 1-5 analogues with substitutions of Gly4 for Leu, D-Gln, Aal or Tal were determined by energy calculations. The above analogues were shown to possess different affinities toward opiate receptors of mu-type. The comparison of low-energy backbone structures of DM, DM 1-5 and its analogues resulted in development of the dermorphin "biologically active" conformation being characteristic of its binding with mu-type receptors. The specific binding of dermorphin to this receptor apparently depends on the conformation of the whole N-terminal pentapeptide. PMID- 2556150 TI - [Antigenic structure of foot-and-mouth virus. IV. Synthesis and immunogenic properties of new fragments of the VP1 protein of food-and-mouth virus strain A22]. AB - A synthesis of new fragments of VP1 protein with the specificity of A22 strain of foot-and-mouth disease virus is described. Immunization with the free 136-152 peptide and KLH-conjugates of the peptides 136-152 and 197-213 induced 60-80% protection of guinea pigs against challenge with the A22 virus. Synthetic peptides corresponding to the 10-24, 50-69 and 175-189 sequences of VP1 did not show any protective activity. We have found that uncoupled peptides 175-189 and 197-213 are able to induce antipeptide antibodies. However, these antibodies did not possess any neutralizing activity. Immunization of animals with the mixture of (136-152)O1K and (175-189)A22 has led to inhibition of the immune response to the (136-152)O1K fragment. PMID- 2556151 TI - Potentiation of the effects of atrial natriuretic factor on the cardiovascular system by amiloride. AB - Amiloride has previously been shown to facilitate receptor binding of atrial natriuretic factor (ANF) to membranes of adrenal cortex and to enhance ANF induced inhibition of steroid secretion in vitro. This interaction of amiloride and ANF also holds true for the cardiovascular system. In precontracted rabbit aortic strips the relaxing effect induced by the combination of ANF (10(-10) mol/l) and amiloride (10(-5) mol/l) was overadditional. The production of cyclic guanosine monophosphate (cGMP), which parallels ANF induced relaxations of vascular strips, was not affected by amiloride alone up to 10(-3) mol/l, but was concentration-dependently increased in the presence of ANF (10(-8) mol/l). In spontaneously hypertensive rats ANF-induced decreases in blood pressure were potentiated by amiloride. Post ischemia reperfusion arrhythmias in isolated rat hearts were reduced by ANF. Amiloride increased this effect. The binding experiments revealed an interaction of amiloride and ANF on the receptor level. Binding of labeled ANF to aortic tissue was concentration-dependently increased by amiloride. Addition of ATP had the opposite effect. Therefore it can be suggested that amiloride and ATP interfere with a mechanism regulating the sensitivity of the vascular ANF-receptor for its ligand regarding binding and signal transforming presumably by a kinase mediated phosphorylation/dephosphorylation process. PMID- 2556152 TI - Spin label studies of erythrocytes during storage of blood. AB - The kinetic behavior of the spin label MAL-6 in the interaction with differently aged human erythrocyte membranes was evaluated by monitoring the rate of disappearance of the room temperature ESR signal due to the MAL-6 spin label added to blood after storage at 4 degrees C or after incubation of red cells at 37 degrees C in a protein-free medium. After 35 days of blood storage or 60 h of erythrocytes incubation at 37 degrees C the decrease of the intensity of the MAL 6 ESR spectra in respect to control samples is markedly enhanced and the correspondent kinetic constants significantly increase. Signal decay of MAL-6 is a further proof that during storage of blood under blood bank conditions or during an artificial ageing of erythrocytes at 37 degrees C, profound modifications occur in the human erythrocyte membrane. PMID- 2556153 TI - Temperature-dependent ESR studies of radical pairs in single crystals of barbituric acid. AB - After irradiation of single crystals of barbituric acid with X-rays at 77 K different types of radical pairs are found, which are composed of only one type of monoradical. The properties of radical pairs of the unpaired electrons are studied using frequency-variable ESR methods at various temperatures. In addition to the radical pairs AD and BC two further pairs A'D' and XY were identified. Measurements of the fine structure parameter D showed a linear temperature dependence in some regions between 77 K and 290 K. At 240 K the radical pairs AD and A'D' changed reversibly into A*D* and this pair remained stable up to 290 K. A level anticrossing effect was observed with the pair AD. The exchange energy J between the singlet state and the triplet state was determined as -(15.1 +/- 0.6) GHz at 77 K and its temperature coefficient as -(3.8 +/- 0.8) x 10(-3)K-1. PMID- 2556154 TI - Desensitization of the beta-adrenergic receptor of the S49 murine lymphoma cell: mechanism of receptor loss and recovery. AB - The effect of prolonged exposure of S49 murine lymphoma cells to (-) isoprenaline on the levels of beta-adrenergic receptors and the stimulatory regulatory component of adenylyl cyclase (Gs) was investigated. Exposure of wild-type S49 cells (WT cells) to isoprenaline for 16 h resulted in an 84% decrease in beta adrenergic receptors and a 78% decrease in catecholamine-stimulated adenylyl cyclase activity. The desensitization of adenylyl cyclase was homologous since no change in stimulation by any other effectors was observed. Similar treatment of cyc- S49 cells, a mutant that lacks Gs, resulted in only a 10-15% receptor loss despite a 50% decrease in catecholamine stimulation of reconstituted adenylyl cyclase activity. By quantitative extraction of Gs from control and isoprenaline treated WT cell membranes and reconstitution into untreated cyc- cell membranes, it was demonstrated that the levels and behaviour of Gs in WT cell membranes were unchanged after isoprenaline treatment, despite a substantial loss of receptors from the cell surface. The beta-adrenergic receptors lost from the cell surface could not be identified in any intracellular membrane fraction. Addition of cycloheximide to the culture medium of WT cells previously treated with isoprenaline resulted in a complete blockade of the restoration of beta adrenergic receptors which was otherwise observed within 16 h after removal of the agonist. It is concluded that, in the S49 cell line, prolonged exposure to agonists results in a loss of beta-adrenergic receptors by a process dependent on the presence of Gs. (ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556155 TI - Is postburn dermal ischaemia enhanced by oxygen free radicals? AB - The accumulated data suggest that oxygen free radicals are actively involved in the pathophysiology of the wound healing process. Since OH. and O2- directly correlate with the release of arachidonic acid and catalyse some of the enzymes participating in its cascade, their involvement in the enhancement of dermal ischaemia in the stasis zone is postulated. Experimental data using topically applied scavengers of superoxide have shown their beneficial effects on the burn wound healing process. PMID- 2556156 TI - [Effect of Rheum tanguticum Maxim ex Balf on cAMP in the central nervous system]. AB - Rabbit model of fever was produced by means of subcutaneous inoculation of pneumocci. Third ventricular intubation and irrigation were carried out in rabbits. Irrigation fluid was collected during the stage of normal temperature, at the peak of fever and when the temperature was lowered by rhubarb. cAMP was detected with RlA in each of the three portions of the irrigated CSF. The result showed that cAMP level was raised during fever and decreased after rhubarb administration. Irrigation of CSF without medication had no significant influence on cAMP level. PMID- 2556157 TI - Formation of superoxide radical and hydrogen peroxide enhanced by trinitrotoluene in rat liver, brain, kidney, and testicle in vitro and monkey liver in vivo. AB - Trinitrotoluene (TNT) increased the formation of adrenochrome from adrenaline and the formation of formaldehyde from methanol in rat liver mitochondria and microsomes in vitro as well as in monkey liver mitochondria and microsomes in vivo. The effects were more prominent at higher TNT concentrations. These findings indicate that TNT enhances the production of superoxide radicals (O2) and hydrogen peroxide (H2O2). The production of O2 was more prominent in systems containing added TNT than in those containing added benzyl viologen. H2O2 production by mitochondria was more pronounced in the liver than in other organs, but its production by microsomes was more pronounced in the brain than in other organs. The results suggest that TNT undergoes cycling reduction which produces oxidative stress. PMID- 2556158 TI - Inability of nitrendipine to protect against gentamicin nephrotoxicity in the rat. AB - Daily sc injection of gentamicin (100 mg/kg) for 4 days produced a significant decrease in the activities of renal cortical Na+,K+-ATPase and alkaline phosphatase. The observed reduction in renal functional enzymatic markers was associated with significant elevation in sphingomyelin, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, phosphatidylcholine, and total phospholipid. Gentamicin significantly decreased the activity of renal phospholipase C. Nitrendipine (25 mg/kg/day) for 7 days po for 4 days alone did not markedly alter the activities of kidney phospholipase C, alkaline phosphatase, and Na+,K+-ATPase or tissue phospholipid levels. Daily administration of nitrendipine for 3 days followed by concurrent treatment of nitrendipine and gentamicin failed to prevent antibiotic-induced renal histopathologic changes, phospholipidosis, or decrease in alkaline phosphatase. However, in rats simultaneously given nitrendipine and gentamicin the activity of Na+,K+-ATPase returned to control values, indicating a selective blocking action for nitrendipine. The inability of nitrendipine to prevent gentamicin-induced renal phospholipidosis or decreases in enzymatic function markers was associated with significantly elevated tissue aminoglycoside levels when compared to values seen in rats given only the antibiotic. Evidence suggests that nitrendipine is not effective in lowering the concentration of gentamicin in renal cortex. The effectiveness of an agent in providing protection against aminoglycoside nephrotoxicity may be associated with the ability of the drug to lower renal gentamicin content. PMID- 2556159 TI - A general rule for ranged series of codon frequencies in different genomes. AB - Information science widely uses descriptions of the distribution of information units (words) according to the frequency of occurrence with the help of a corresponding ranged series, i.e., the sequence of occurrence frequencies p1, p2, ..., pr as taken in decreasing order. A model called the Zipf rule or Zipflaw is the most commonly used. In this model pr is inversly proportional to a certain degree of range r: pr = C/r2 (C, z greater than 0). Upon analysis, the correspondence of codon distribution and the Zipf model is found unsatisfactory. The distribution of letters (in English and some other languages) by the occurrence frequency does not obey the Zipf rule either. A new model is proposed for a similar distribution in which pr = C.(ln(n + 1)-ln r), where n is the quantity of various symbols (codons). This dependence is approximated by a straight line not in the co-ordinate system (ln r, ln p), like the Zipf model, but in the (ln r, p) system of co-ordinates. It is shown on the basis of statistical criteria that this model is in good agreement with the ranged series of codon frequencies for the best-studied genoms to date. This result may be regarded as an additional reason in favor of the codon-letter analogy (not the codon-word analogy) in genetic texts. PMID- 2556160 TI - Control region of SV40 minichromosomes is preferentially cleaved by single-strand specific S1 nuclease. AB - We have analysed S1 sensitivity of SV40 minichromosomes isolated from the nuclei of infected cells at the late stage of infection. We show that a fraction of purified minichromosomes is sensitive towards double-strand cleavage by S1 nuclease. The pattern of specific cleavage reminiscent of that found for subcloned fragment under supercoiling is superimposed upon apparently random double-strand cuts along the entire regulatory region. Therefore, the cleavage sites are not exclusively confined to the regions with the reported alternate DNA conformation. PMID- 2556161 TI - Hamster liver cholinephosphotransferase and ethanolaminephosphotransferase are separate enzymes. AB - CDP-choline:1,2-diacylglycerol cholinephosphotransferase (EC 2.7.8.2) and CDP ethanolamine:1,2-diacylglycerol ethanolaminephosphotransferase (EC 2.7.8.1) are microsomal enzymes that catalyze the final steps in the syntheses of phosphatidylcholine and phosphatidylethanolamine via the CDP-choline and CDP ethanolamine pathways, respectively. Both enzyme activities were cosolubilized from hamster liver microsomes by Triton QS-15. Limited separation of these two activities was achieved by ion-exchange chromatography. The partially purified phosphotransferases displayed a higher sensitivity than microsomal phosphotransferases towards exogenous phospholipids and showed an absolute requirement for divalent cations. Upon purification, cholinephosphotransferase was more stable to heat treatment than ethanolaminephosphotransferase. The two enzymes exhibited distinct pH optima and responded differently to exogenous phospholipids. Our results clearly indicate that cholinephosphotransferase and ethanolaminephosphotransferase are separate enzymes. PMID- 2556162 TI - Characterization of the fragmented forms of calcineurin produced by calpain I. AB - Calcineurin, a calmodulin-stimulated phosphatase from bovine brain, was hydrolyzed by calpain I from human erythrocytes. In the absence of calmodulin, calpain rapidly transformed the 60-kilodalton (kDa) catalytic subunit of calcineurin into a transient 57-kDa fragment and thereafter a 43-kDa limit fragment. In the presence of calmodulin, the 60-kDa subunit was sequentially proteolyzed to a 55-kDa fragment and then a 49-kDa fragment. Upon proteolysis in the absence or presence of calmodulin, the p-nitrophenyl phosphatase activity (assayed in the presence of calmodulin) was increased by 300%. The 43- and the 49 kDa fragments were found to (i) remain associated with the small subunit (17 kDa), (ii) have lost the ability to bind and to be activated by calmodulin, and (iii) have phosphatase activity that was still stimulated by Mn2+ or Ni2+. The 43 + 17-kDa form had similar Km values for various substrates, but the Vmax values were increased compared with the native enzyme. It is proposed that (i) a 43-kDa core segment of the 60-kDa subunit of calcineurin contained the catalytic domain, the small subunit-binding domain, and the metal ion (Mn2+ and (or) Ni2+) binding site; and (ii) two distinct types of inhibitory domains exist near the end(s) of the large subunit, one of which is calmodulin regulated, while the other is calmodulin independent. PMID- 2556163 TI - Pyruvate fermentation by Clostridium acetobutylicum. AB - Clostridium acetobutylicum ATCC 824 using pyruvate as the sole carbon source produced mainly acetate and butyrate as end products of fermentation. Acetate and butyrate kinase activities were higher in cells growing in the presence of pyruvate than glucose, whereas the level of the acetoacetate decarboxylase, an enzyme involved in solvent formation, was lower. Similar activities of glyceraldehyde-3-phosphate dehydrogenase were found in cells grown in pyruvate and glucose mediums. The transfer of C. acetobutylicum from pyruvate to glucose medium suggested that pyruvate represses the "solventogenesis." PMID- 2556164 TI - Epidemiology of human papillomavirus (HPV) infections and their associations with genital squamous cell cancer. Review article. AB - Reliable assessment of the epidemiology of genital HPV infections is hamphered by a number of technical problems. Because of the lack of tissue-culture systems, methods based on morphological approaches (colposcopy, cytology and histopathology) play a central role in HPV diagnosis. Even DNA-hybridization techniques and the recently introduced DNA amplification with PCR are extremely difficult to standardize, and are thus subject to major interlaboratory variation. Further confusion in the field is created by the complex biological behaviour of HPV infections. As established by the long-term prospective follow up study of over 500 women which has been running in Kuopio since 1981, clinical progression and regression are significantly related to the grade of the lesion at the time of diagnosis (p less than 0.00001, and p = 0.0005, respectively), as well as to the type of HPV (p = 0.0012). Most importantly, however, genital HPV infections seem to run an extremely fluctuating course, passage from manifest to subclinical or latent infection being frequently encountered in individual patients when examined at 6-month intervals over prolonged periods. This explains the significantly divergent prevalence figures reported in different series (ranging from 2% to 80%), which are completely dependent on the technique used to analyse the presence of HPV, i.e. whether a) PAP smear, b) biopsy, c) DNA hybridization, or d) PCR amplification. The first two are capable of disclosing only manifest (clinical) infections, the latter two also the latent ones. In an unselected population of 22-year-old Finnish females, the prevalence of clinical HPV infections was about 3 per cent, and the adjusted annual incidence was 8.0 per cent. According to estimates of the life-time risk, up to 79% of Finnish females will contract at least one HPV infection between the ages 20 to 79 years. When related to the long-term trends in invasive cervical cancer in Finland, it is evident that this 79% life-time risk of becoming HPV-infected or even the observed 15% clinical progression rate for HPV infections in the prospective follow-up study by no means signifies an identical risk of developing cervical cancer (i.e. 0.79 x 0.15 = 11%). It seems likely that in countries where mass screening programmes exist (and precancer lesions are traced), the high prevalence of HPV infections is not necessarily reflected as an increased prevalence of invasive cervical carcinomas. The distinction of lesions at risk for malignant transformation from those regressing spontaneously will have major implications in therapeutic considerations of genital HPV infections. PMID- 2556165 TI - Effects of sodium nitroprusside on blood glucose concentration, B-cell morphology and islet glutamate dehydrogenase activity in mice. AB - Mice injected with sodium nitroprusside (NaNP) exhibited a marked, transient hyperglycemia and enhanced activity of glutamate dehydrogenase in the pancreatic islets. Ultrastructurally, the islet B-cells of NaNP-treated mice showed expanded granular endoplasmic reticulum, prominent Golgi complex, increased amount of secretory granules, mitochondrial enlargement and vacuolation, and mitochondrion secretory granule complexes. Stereological analyses disclosed increased volume of endoplasmic reticulum, Golgi complex and mitochondria, and increased number of secretory granules in the B-cells 1 h after the injection of NaNP. Isolated mouse islets exposed to NaNP showed stimulated activity of glutamate dehydrogenase both in the presence of 2 and 18 mM glucose, whereas the release of insulin was stimulated at 2 mM glucose, but inhibited at 18 mM glucose. The observations demonstrate that NaNP induces transiently altered structure and function in mouse islet B-cells. PMID- 2556166 TI - New receptor for human plasminogen on gram positive cocci. AB - 180 bacterial strains representing 17 different species of gram positive cocci were tested for the ability to interact with human plasminogen. Receptors for plasminogen could be detected on 23/24 strains of S. pyogenes, 15/15 strains of S. equisimilis, 14/16 strains of human group G streptococci and 14/14 strains of S. pneumoniae. Eight of nineteen strains representing five species of alpha hemolytic streptococci were also positive. S. equisimilis demonstrated the highest uptake with a median value of 58 per cent (20%-67%). On the other hand, all strains of S. agalactiae, the majority of S. faecalis and all S. aureus, S. epidermidis and S. saprophyticus strains tested were negative. The concentration of unlabelled plasminogen causing a 50 per cent reduction of bound tracer was between 50 and 150 mM. These estimates of the dissociation constant confirmed the specific nature of the interaction. Binding of plasminogen could be blocked by addition of plasmin-aprotinin complex, suggesting that plasminogen and plasmin bind to the same receptor. Binding was also blocked by the plasminogen fragment kringle 1-3, but not by miniplasminogen, a fragment containing kringle 5 and the B-chain region. As streptokinase interacts mainly with the B-chain of plasmin it is clear that the bacterial receptor for plasminogen is not identical to streptokinase. PMID- 2556167 TI - Adult Wilms' tumor. Role of combined modality treatment. AB - Adults Wilms' tumor, unlike its pediatric counterpart, is extremely rare and highly lethal. Less than 200 cases have been reported in the world literature and a large proportion of these may not have been true Wilms' tumors. We report 3 cases seen at our institution who were treated with a multimodality approach, all three remaining alive without evidence of active disease at 6 years, 2.25 years and 8 months respectively from diagnosis. We also review results of multimodality management, emphasizing the poor outcome for stage III and IV tumors in spite of multi-agent chemotherapy. Alternative drugs, such as cisplatinum may have a role in management. Finally, we propose that prophylactic pulmonary irradiation may have a place in advanced stages of adult Wilms' tumor. PMID- 2556169 TI - Small cell lung cancer. PMID- 2556168 TI - The release of chromogranin A and B like activity from human lung cancer cell lines. A potential marker for a subset of small cell lung cancer. AB - Human small cell lung cancer (SCLC) is in vivo and in vitro characterized by a heterogeneous spectrum of neuroendocrine markers. The non-SCLC group is deprived of these markers, or expresses them in low quantities. In this paper we report on the release to the culture medium of neuroendocrine associated proteins, chromogranin A and B like activity (CABLA). The culture medium from three out of five SCLC cell lines and in one/five non-SCLC cell line contained significant levels of CABLA. Normal diploid foreskin fibroblasts and a histiocytic lymphoma cell line were deprived of CABLA production. The presence of CABLA in both SCLC and non-SCLC further stress their common histogenetic origin. The CABLA values were partly unrelated to other neuroendocrine markers. Determinations of CABLA could thus be a potential and valuable marker for a subset of SCLC. PMID- 2556170 TI - HIV-related lentiviruses of nonhuman primates. PMID- 2556171 TI - Oral medicine in practice: viral infection. AB - The fourth article in this series deals with viral infections which may present in the dental surgery. Two groups of viruses, herpes and coxsackie, are responsible for the majority of viral conditions involving the orofacial region which present to the dentist. Mucosal ulceration is the most frequent clinical presentation, although viruses may also be responsible for salivary gland swelling or epithelial hyperplasia. Diagnosis of viral infection is important, since treatment which can alleviate symptoms and reduce the likelihood of spread of infection is available. In addition, the recognition of intra-oral viral infection can have serious implications, since it may be an indication of underlying conditions such as leukaemia, HIV infection or child abuse. PMID- 2556173 TI - Nabilone as effective therapy for intractable nausea and vomiting in AIDS. PMID- 2556172 TI - Lisinopril dose-response relationship in essential hypertension. AB - 1. This was a multicentre, double-blind, parallel study in 216 patients with mild to moderate (supine diastolic blood pressure = 95-115 mm Hg) essential hypertension. 2. After a 4-week placebo washout, patients were randomized to placebo or lisinopril 1.25, 5.20 or 80 mg once daily for 6 consecutive weeks. Supine and erect blood pressure was measured 24 h postdose at the end of weeks 2, 0, 2, 4, and 6. 3. There was a linear dose-response relationship for both supine and erect blood pressure. Diastolic blood pressure reductions in the lisinopril 20 and 80 mg day-1 groups were significantly greater than in the placebo or lisinopril 1.25 and 5 mg day-1 groups. 4. Lisinopril, at doses up to 80 mg day-1, was well tolerated. PMID- 2556174 TI - Comparative intragastric pH profile of Algicon versus Gaviscon. PMID- 2556175 TI - Comparative study of Algicon suspension and magnesium trisilicate mixture in the treatment of reflux dyspepsia of pregnancy. PMID- 2556176 TI - Comparative study of Algicon versus Gaviscon in symptomatic gastro-oesophageal reflux. PMID- 2556177 TI - Algicon in the management of gastro-oesophageal reflux. PMID- 2556178 TI - Characteristics of alveolar cells and soluble components in bronchoalveolar lavage fluid from non-smoking aluminium potroom workers. AB - Aluminium potroom workers have been reported to develop severe pneumoconiosis and bronchial hyperreactivity. The influence of inhalation of aluminium oxide and fluorides on the alveolar milieu was studied by bronchoalveolar lavage (BAL) in 14 male non-smoking potroom workers; 28 non-smoking healthy volunteers served as controls. The total numbers, concentrations, and proportions of various alveolar cells did not differ between the groups. The concentrations of albumin and fibronectin in BAL fluid were significantly higher (p less than 0.01 for both) in the exposed workers, reflecting an increased alveolar capillary permeability and an activation of alveolar macrophages (AMs). The concentration of angiotensin converting enzyme, another AM marker, was, however, decreased (p less than 0.01) in the workers. The concentration of hyaluronan, a fibroblast marker, did not differ between the groups. AMs from workers had a decreased capacity (p less than 0.05) to interact with yeast C3b particles but not to ingest them. The expression of HLA-DR and OKM1 on the cell surfaces of AMs were equal in the two groups. The BAL findings were not accompanied by restrictive lung disease in the workers. The fact that only a discrete alveolitis was found in the potroom workers may be due to a low grade of exposure to alumina and fluorides and to frequent use of respiratory protection equipment. PMID- 2556179 TI - Inorganic pyrophosphate in mitochondrial metabolism. PMID- 2556180 TI - Long-chain fatty acids act as protonophoric uncouplers of oxidative phosphorylation in rat liver mitochondria. AB - The effect of long-chain fatty acids (LCFA) on respiration and transmembrane potential (delta psi) in the resting state, and the rate of delta psi dissipation [d delta psi/dt)i) was investigated with oligomycin-inhibited rat liver mitochondria using succinate (plus rotenone) as substrate. The results obtained were compared with those of classical protonophores such as 2,4-dinitrophenol (DNP) and 4,5,6,7-tetrachloro-2-trifluoromethylbenzimidazole (TTFB). The effects of oleate or palmitate and that of DNP or TTFB on respiration and delta psi can be described by a common force-flow relationship. These facts all in all are not compatible with a decoupler-type uncoupling mechanism of LCFA; still, they indicate that the latter are protonophores. Moreover, the oleate-induced increase in the rate of delta psi dissipation closely correlates with that in respiration, suggesting that the uncoupling activity and the protonophoric activity of LCFA are interrelated. Carboxyatractyloside (CAT) exerted only a small inhibitory effect on oleate-induced respiration and delta psi dissipation, indicating that the adenine nucleotide translocase contributes to the uncoupling effect of LCFA to a minor extent only. Proton transport through the lipid region of the membrane as mediated by permeation of the protonated and deprotonated forms of LCFA is interpreted as the main process of the uncoupling of LCFA. PMID- 2556181 TI - The effect of pH and temperature on the reaction of fully reduced and mixed valence cytochrome c oxidase with dioxygen. AB - The reaction of fully reduced and mixed-valence cytochrome oxidase with O2 has been followed in flow-flash experiments, starting from the CO complexes, at 428, 445, 605 and 830 nm between pH 5.8b and 9.0 in the temperature range of 2-40 degrees C. With the fully reduced enzyme, four kinetic phase with rate constants at pH 7.4 and 25 degrees C of 9 x 10(4), 2.5 x 10(4), 1.0 x 10(4) and 800 s(-1), respectively, are observed. The rates of the three last phases display a very small temperature dependence, corresponding to activation energies in the range 13-54 kJ x mol(-1). The rates of the third and fourth phases decrease at high pH due to the deprotonation of groups with pKa values of 8.3 and 8.8, respectively, but also the second phase appears to have a small pH dependence. In the reaction of the mixed-valence enzyme, three kinetic phases with rate constants at pH 7.4 and 25 degrees C of 9 x 10(4), 6000 and 150 s(-1), respectively, are observed. The third phase only has a small temperature dependence, corresponding to an activation energy of 20 kJ x mol(-1). No pH dependence could be detected for any phase. Reaction schemes consistent with the experimental observations are presented. The pH dependencies of the rates of the two final phase in the reaction of the fully reduced enzyme are proposed to be related to the involvement of protons in the reduction of a peroxide intermediate. The temperature dependence data suggest that the reorganization energies and driving forces are closely matched in all electron transfer steps with both enzyme forms. It is suggested that the slowest step in the reaction of the mixed-valence enzyme is a conformation change involved in the reaction cycle of cytochrome oxidase as a proton pump. PMID- 2556182 TI - Cytochrome c-551 from the thermophilic bacterium PS3 grown under air-limited conditions. AB - A small-sized c-type cytochrome, designated cytochrome c-551, was prepared from membrane fraction of the thermophilic bacterium PS3 grown under air-limited conditions by extraction with cholate, precipitation with polyethylene glycol, and successive chromatographies with DEAE-cellulose and Sephacryl S-200 in the presence of a detergent. The purified sample contained approximately 1 mol of heme c per 10,000 g protein; it showed absorption bands at 551, 522 and 416 nm upon reduction, and a Soret peak at 409 nm upon oxidation. This cytochrome showed a single band of 10 kDa on polyacrylamide gel electrophoresis with sodium dodecyl sulfate. The isoelectric point of this cytochrome c-551 was pH 4.0. Cytochrome c 551 was suggested to play an important role in the respiratory chain with a terminal oxidase cytochrome o, which is produced under air-limited conditions, since cytochrome c-551 could mediate electron transfer between cytochrome bc1(b6f) complex and cytochrome o, showing quinol oxidase activity. PMID- 2556183 TI - The F1F0-ATPase of Escherichia coli. The substitution of glycine by valine at position 29 in the c-subunit affects function but not assembly. AB - Site-directed mutagenesis has been used to construct two mutations within the uncE gene, coding for the c-subunit of the F1F0-ATPase, resulting in the substitution of Gly-29 by Val and Gly-18 by Leu. The strain carrying the Gly-29-- -Val substitution is unable to grow on succinate as sole carbon source and possesses an uncoupled growth yield, while the strain carrying the Gly-18----Leu substitution possesses a wild-type phenotype. Membranes prepared from the strain carrying the Gly-29----Val substitution possess low levels of ATPase activity and are proton-impermeable. The F1-ATPase activity of this strain was found to be inhibited by approx. 75% when bound to the membrane. These results are discussed in relation to a previously proposed model for the F0 (Cox, G.B., Fimmel, A.L., Gibson, F. and Hatch, L. (1986) Biochim. Biophys. Acta 849, 62-69). PMID- 2556184 TI - [Study of the complex formation between alpha- and beta- forms of NAD-kinase and glutamate dehydrogenase and the effect of metabolites on enzyme interaction]. AB - The alpha- and beta-forms of rabbit liver NAD kinase were found to differ significantly in terms of their ability to form complexes with glutamate dehydrogenase. The beta-form of the enzyme was shown to form a more stable complex with glutamate dehydrogenase (Kd = 1.5.10(-8) M), whereas the Kd value for the alpha-form is 2.9.10(-7) M. Using two independent methods, it was shown that in the absence of effectors 40% of the beta-form of NAD kinase and up to 20% of the alpha-form are bound to glutamate dehydrogenase. The substrates of NAD kinase markedly activate the complex formation only in the case of the alpha-form of the enzyme. The time needed for this process is also reduced in the presence of the substrates. PMID- 2556186 TI - [The mechanism of rat liver fructose-1,6-bisphosphate inhibition by fructose-2,6 bisphosphate]. AB - It was found that a decrease in the activating cation (Mg2+) concentration below [A]0.5 causes the disappearance of cooperativity of the fructose 1.6 bisphosphatase substrate binding sites induced by high fructose 2.6-bisphosphate concentrations without any significant alteration in the extent of the enzyme inhibition. Under these conditions, a competitive type of inhibition (with respect to the substrate) is transformed into a non-competitive type with an increase in the fructose 2.6-bisphosphate concentration. The data obtained confirm the viewpoint that fructose 2.6-bisphosphate binds to the enzyme at two distinct sites, a catalytic and an allosteric ones, differing in their affinity for the inhibitor. It is supposed that the interaction between the allosteric fructose 2.6-bisphosphate binding site and the activator site occupied by Mg2+ is necessary for the cooperative response of the enzyme to the substrate. PMID- 2556185 TI - [3',5'-cyclic nucleotide phosphodiesterase from human brain]. AB - 3':5'-Cyclic nucleotide phosphodiesterase was isolated from human brain and characterized. After the first stage of purification on phenyl-Sepharose, the enzyme activity was stimulated by Ca2+ and micromolar concentrations of cGMP. High pressure liquid chromatography on a DEAE-TSK-3SW column permitted to identify three ranges of enzymatic activity designated as PDE I, PDE II and PDE III. Neither of the three enzymes possessed a high selectivity for cAMP and cGMP substrates. The catalytic activity of PDE I and PDE II increased in the presence of Ca2+-calmodulin (up to 6-fold); the degradation of cAMP was decreased by cGMP. The Ca2+-calmodulin stimulated PDE I and PDE II activity was decreased by W-7. PDE I and PDE II can thus be classified as Ca2+-calmodulin-dependent phosphodiesterases. With cAMP as substrate, the PDE III activity increased in the presence of micromolar concentrations of cGMP (up to 10-fold), Ca2+ and endogenous calmodulin (up to 2-3-fold). No additivity in the effects of saturating concentrations of these compounds on PDE III was observed. Ca2+ did not influence the rate of cGMP hydrolysis catalyzed by PDE III. In comparison with PDE I and PDE II, the inhibition of PDE III was observed at higher concentrations of W-7 and was not limited by the basal level of the enzyme. These results do not provide any evidence in favour of the existence of several forms of the enzyme in the PDE III fraction. The double regulation of PDE III creates some difficulties for its classification. PMID- 2556188 TI - [Phorbol ester blocks the coupling of GTP-binding protein with receptor controlled calcium channels]. AB - Using the intracellular Ca2+-specific indicator, Quin 2, it was demonstrated that an addition to platelet suspensions of the GTP-binding protein activator, sodium fluoride, stimulates the Ca2+ and Ba2+ influx from the incubation medium into the cytoplasm via receptor-operated Ca2+ channels (Ca-ROC). The fluoride-induced Ca2+ influx is blocked by the protein kinase C activator, phorbol myristate acetate as well as by the platelet adenylate cyclase activator, prostaglandin E1. A two dimensional electrophoretic analysis of platelet phosphoproteins revealed that the phorbol ester enhances the phosphorylation of proteins with molecular masses of about 20 and 40 kDa. The experimental results suggest that the participation of the GTP-binding protein in the receptor coupling to Ca-ROC. The mechanism of the blocking effect of phorbol esters and prostaglandin E1 on Ca-ROC consists in an impaired coupling of these channels to the GTP-binding protein that activates them. PMID- 2556187 TI - [The blocking effect of aliphatic hydrocarbons on Ca2+ leakage channels formed by aggregates of Ca2+-ATPase of the sarcoplasmic reticulum]. AB - The effects of aliphatic hydrocarbons within the liposomes on the Ca2+ transport function of isolated sarcoplasmic reticulum (SR) membranes of rabbit skeletal muscle, vesiculate preparation of Ca2+ dependent ATPase and proteoliposomes reconstituted from Ca2+-ATPase and egg phosphatidylcholine, were studied. It was shown that liposomes prepared from dipalmitoyl phosphatidylcholine containing aliphatic hydrocarbons increase 2 to 3 times Ca2+ accumulation by Ca2+-dependent ATPase from rabbit skeletal muscle SR. Ca2+ transport by SR vesicles increases in the presence of hydrocarbons by 15--20%. The activating effect of hydrocarbons on Ca2+ transport by proteoliposomes depends on the lipid/protein ratio. The proteoliposomes with a high lipid/protein ratio are practically insensitive to the effects of hydrocarbons. It was suggested that activation of Ca2+ transport by hydrocarbons is due to blocking of Ca2+ leakage channels formed during the aggregation of Ca2+-ATPase molecules. Treatment of membranes by formaldehyde results in the oligomerization of Ca2+-ATPase and decreases 2--4-fold the ATP dependent accumulation of Ca2+. Subsequent addition of decane restores Ca2+ transport practically completely. PMID- 2556189 TI - Circadian variation of platelet 3H-imipramine binding, platelet serotonin content, and plasma cortisol in healthy volunteers. AB - Platelet 3H-imipramine binding, platelet serotonin content, and plasma cortisol were measured in 10 healthy female volunteers. Samples were taken on 5 occasions with a 6-hr interval during a 24-hr period within 1 week in May. Although there were individual variations in the biochemical parameters over a 24-hr period, we could not establish a clear common circadian rhythm for all subjects in platelet 3H-imipramine binding and platelet serotonin concentrations. During the night, there was a significant decrease in Bmax values and a slight, but not significant, increase in serotonin concentrations. A distinct circadian rhythm was obtained for plasma cortisol, with lowest values during the night. PMID- 2556190 TI - Lithium, brain, and RBC NMR parameter T-1. PMID- 2556191 TI - Catecholestrogens inhibit basal and luteinizing hormone-stimulated androgen production by porcine thecal cells. AB - It has been shown recently that catecholestrogens are produced by cultured porcine granulosa and thecal cells, and that they influence porcine granulosa cell steroidogenesis in a similar manner to estradiol-17 beta (E2). The present studies were performed to determine if catecholestrogens also play a role in the regulation of porcine thecal cell steroidogenesis and to compare their actions to those of E2. Thecal cells were obtained from prepubertal gilts and cultured in a serum-free medium for 48 h. Thecal cell androstenedione production under basal and luteinizing hormone (LH)-stimulated conditions was significantly inhibited by adding E2 or catecholestrogens to the culture medium. Treatment of basal and LH stimulated cultures with increasing concentrations of E2 or catecholestrogens (0.1-10 micrograms/ml) caused a dose-and time-dependent inhibition of androstenedione production. The inhibitory effect of the catecholestrogens, but not of E2, was enhanced when the cultures contained the catechol-O-methyl transferase inhibitor, U-0521. Studies to determine the mechanism(s) of action of the catecholestrogens showed that E2 and catecholestrogen actions are exerted at a site(s) distal to cyclic adenosine 3'5' monophosphate (cyclic AMP) generation, because neither agent affected the basal or LH-stimulated accumulation of extracellular cyclic AMP, while causing a significant inhibition of androstenedione production. E2 or catecholestrogen treatment also inhibited androstenedione production stimulated by prostaglandin E2 and dibutyryl cyclic AMP. In addition, both E2 and catecholestrogen treatment significantly decreased basal and LH-stimulated 17 alpha-hydroxyprogesterone production, while significantly increasing pregnenolone production. Progesterone production in the presence of E2 or catecholestrogens showed small but statistically insignificant increases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556192 TI - Gonadotropin-releasing hormone binding sites in ovaries of normal cycling and persistent-estrus rats. AB - The gonadotropin-releasing hormone (GnRH) binding capacity in ovaries and pituitaries of normal cycling rats at different stages of the estrous cycle and in ovaries of persistent-estrus rats was measured using radioligand-receptor assay (RRA). Persistent estrus was induced either by neonatal administration of testosterone propionate (1.25 mg s.c.) on the second day of life or by a hypothalamic suprachiasmatic frontal cut made with Halasz' knife. All animals were killed during the critical period (1400-1600 h), and GnRH receptor was assayed. GnRH receptor levels in both ovaries and pituitaries changed during the estrous cycle. The total number of ovarian GnRH binding sites was significantly higher in proestrus than in diestrus 1, the stage in which the lowest level was found. When binding sites were expressed in fmol/mg ovary, the highest level was observed in diestrus 2; however, no changes were observed during the estrous cycle when GnRH binding sites were expressed as fmol/mg protein. Changes noted were very similar to those demonstrated in pituitary GnRH receptors in our present and previous experiments. Higher levels of pituitary binding sites were found in diestrus 2 and proestrus than in estrus and diestrus 1. The changes in the GnRH receptor levels were more striking in the pituitary than in the ovaries. It appears that the total number of ovarian GnRH binding sites was not altered in either of the two persistent-estrus groups, but that their concentration was significantly higher (expressed in fmol/mg ovary or fmol/mg protein) than on any day during the estrous cycle.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556193 TI - Some aspects of calcium regulation in cell biology. AB - The roles of intracellular calcium in the regulation of cell metabolism and cell membrane permeability are highlighted with examples taken from recent studies. PMID- 2556194 TI - Model membrane partition ESR study in the presence of alpha-tocopherol by a new spin probe. AB - The effect of alpha-tocopherol (alpha T) on partitioning and fluidity changes occurring in phospholipid liposomes have been investigated by monitoring the X band ESR spectrum of the high resolution amphiphilic spin probe perdeutero-di-t butyl nitroxide (PDDTBN), which partitions in the lipid and water phase of liposomes, showing all the three resonances from each phase well resolved. PMID- 2556195 TI - Principles of DNA cloning. PMID- 2556196 TI - Cytomegalovirus in blood from blood donors. PMID- 2556197 TI - The use of a non-linear regression approach for the analysis of the ouabain-K+ interaction with (Na+ + K+)-ATPase from guinea pig and rat hearts. AB - 1. The interaction between ouabain and K+ and their effects on (Na+ + K+)-ATPase activity were studied using microsomes from guinea pig and rat heart. 2. Microsomes were incubated in the presence of various concentrations of K+ and ouabain and ATPase activity was estimated by measuring the inorganic phosphate liberated. The experimental data were analyzed statistically by micro-computer, using a non-linear regression program based on the steepest descent technique. 3. The experimental data were best fitted by a model which assumes that ouabain acts like a mixed inhibitor with respect to the apparently cooperative K+ activation of (Na+ + K+)-ATPase. This quantitative approach provided estimates (with approximate standard deviations) of all the parameters involved in the model. 4. The inhibition constant for the uncompetitive term of the effect was 7- to 9-fold higher than the inhibition constant for the competitive term for both the guinea pig and rat heart preparations. 5. The present results indicate that graphical analyses are helpful for illustrative purposes but suggest that a computerized, non-linear regression program simultaneously analyzing all the non-linearized data should be used to quantify the complex kinetic parameters and to discriminate objectively among possible models. PMID- 2556198 TI - Cycloleucine blocks NMDA responses in cultured hippocampal neurones under voltage clamp: antagonism at the strychnine-insensitive glycine receptor. AB - 1. Radioligand binding studies have demonstrated that the neutral amino acid cycloleucine may act as a competitive antagonist at the glycine modulatory site on the N-methyl-D-aspartate (NMDA) receptor complex. In the present study, we examined the effects of cycloleucine on NMDA-evoked inward current responses in dissociated hippocampal neuronal cultures using the whole cell voltage-clamp technique. 2. In the presence of 1 microM glycine, cycloleucine caused a reversible, dose-dependent inhibition of NMDA responses with an IC50 of 24 microM. An increase in glycine to 100 microM resulted in a shift to the right of the cycloleucine concentration-effect curve (IC50, 1.4 mM). However, with cycloleucine concentrations less than or equal to 100 microM, a fraction of the block could not be overcome by glycine even at concentrations as high as 1 mM. 3. The cycloleucine block was unaffected by shifts in the holding potential (-60 to +60 mV), and there was no effect of cycloleucine on the reversal potential of the NMDA-evoked current. 4. Cycloleucine failed to effect kainic acid- and quisqualic acid-evoked currents at concentrations which inhibited NMDA responses. 5. We conclude that cycloleucine is a potent and selective antagonist of NMDA-receptor mediated responses. Although this effect occurs in part via competitive antagonism at the glycine modulatory site, the cycloleucine block cannot be completely reversed by glycine indicating an interaction with an additional site on the receptor-channel complex. PMID- 2556200 TI - Augmentation and subsequent attenuation of Ca2+ current due to lipid peroxidation of the membrane caused by t-butyl hydroperoxide in the rabbit sinoatrial node. AB - Cellular electrophysiological effects of membrane lipid peroxidation by t-butyl hydroperoxide (TBH) were studied in the rabbit sinoatrial (SA) node. Superfusion for 1-5 min with 300 microM TBH caused an initial increase and subsequent decrease in the spontaneous firing frequency of the SA node. Voltage clamp experiments revealed that TBH initially enhanced but later blocked the Ca2+ current. Thus, membrane lipid peroxidation appears to accelerate and then suppress physiological automaticity by causing biphasic changes in the Ca2+ current. PMID- 2556201 TI - Pharmacological characterization of postjunctional alpha-adrenoceptors in cerebral arteries from the sheep. AB - 1. The responsiveness to noradrenaline was characterized in cerebral arteries from the sheep, since this species was large enough to permit a comparison of arteries from different parts of the cerebral vasculature. 2. Noradrenaline caused contraction of the basilar artery, middle cerebral artery and small pial arteries by stimulation of alpha 1-adrenoceptors. 3. The maximum contraction to noradrenaline was small in the basilar artery (28% of the 5-hydroxytryptamine (5 HT) maximum) but larger in the middle cerebral artery (78% of the 5-HT maximum) and pial artery (92% of the 5-HT maximum) of the sheep. 4. Cocaine (10 microM) potentiated noradrenaline-induced contractions in the sheep middle cerebral artery but not in the sheep basilar artery. 5. The noradrenaline contraction, relative to the 5-HT contraction, was not affected by removal of the endothelium in either the sheep basilar or middle cerebral artery. 6. The results showed a variation within the sheep cerebral vasculature in the response to noradrenaline which cannot be explained by regional differences in alpha-adrenoceptor subtypes, noradrenaline uptake mechanisms or endothelial function. PMID- 2556199 TI - Delta-opioid mediated inhibitions of acute and prolonged noxious-evoked responses in rat dorsal horn neurones. AB - 1. The effects of a selective delta-opioid agonist Tyr-D-Ser(Otbu)-Gly-Phe-Leu Thr (DSTBULET) were examined on the C- and A beta-evoked responses of convergent dorsal horn neurones in the halothane anaesthetized, intact rat. 2. Intrathecal DSTBULET produced selective dose-dependent inhibitions of electrically-evoked C fibre responses of both superficial and deep neurones. A near-complete inhibition of 83 +/- 5% followed 100 micrograms of DSTBULET and the ED50 was 9 micrograms (13.5 nmol). Inhibitions were antagonised by intrathecal naloxone and ICI 174,864 but were not antagonised by pretreatment with intrathecal beta-funaltrexamine at a dose that blocked mu-opioid effects. By contrast, DSTBULET produced excitations of electrically-evoked responses of cells recorded in a zone intermediate between the superficial and deep neurones. 3. DSTBULET (50 micrograms) was also tested on the more prolonged noxious neuronal response produced by subcutaneous formalin (5%, 50 microliters) into the receptive field. DSTBULET profoundly inhibited the response to formalin. Pretreatment with ICI 174,864 before DSTBULET antagonised the effects of the delta-agonist on the formalin response. 4. The full peptidase inhibitor kelatorphan, known to protect endogenous enkephalins, was also tested on the formalin response. The intrathecal administration of 50 micrograms kelatorphan has previously been shown to inhibit electrically-evoked C fibre resonses of dorsal horn neurones and to be antagonised by ICI 174,864. The same dose of kelatorphan inhibited the formalin response in the present study. 5. From this study it appears that the delta-opioid agonist DSTBULET can produce profound inhibitions of the responses of convergent neurones to nociceptive afferent inputs. Furthermore, activation of delta-opioid receptors either by DSTBULET, or by protection of endogenous enkephalins with kelatorphan, can inhibit a more prolonged chemically-evoked nociceptive input onto these dorsal horn neurones. PMID- 2556202 TI - Effects of the putative antagonists phaclofen and delta-aminovaleric acid on GABAB receptor biochemistry. AB - 1. Phaclofen and delta-aminovaleric acid (delta-AVA) have been reported to be antagonists at gamma-aminobutyric acidB (GABAB) receptors. Phaclofen, delta-AVA and related compounds were examined for potency and specificity at GABAB and GABAA receptors in rat cortical membranes labelled with [3H]-(-)-baclofen and [3H]-muscimol, respectively. Additionally phaclofen and delta-AVA were examined in two functional tests of central GABAB activity in rat cortical slices, namely the inhibition of forskolin-stimulated cyclic AMP accumulation, and the potentiation of isoprenaline-stimulated cyclic AMP accumulation. 2. delta-AVA (IC50 = 11.7 microM) was 20 fold more potent than phaclofen (IC50 = 229 microM) on GABAB receptor binding. All compounds possessing a phosphonic acid group, including phaclofen, which were active at GABAB receptors were inactive at GABAA receptors, while delta-AVA was equally potent at both receptors. Several compounds exhibited Hill coefficients of less than unity in displacing [3H]-(-) baclofen binding. 3. (-)-Baclofen inhibited forskolin-stimulated cyclic AMP accumulation (IC50 = 7.9 microM) but this effect was not stereospecific. Phaclofen (1 mM) was inactive against this inhibition but produced a potentiation of the forskolin effect. delta-AVA (1 mM) failed to antagonize the effect of baclofen; rather it mimicked baclofen. 4. (-)-Baclofen (10 microM) potentiated isoprenaline-stimulated cyclic AMP accumulation, an effect antagonized by phaclofen (1 mM). delta-AVA (1 mM) may be a weak antagonist but also potentiated basal cyclic AMP accumulation. 5. We conclude that neither delta-AVA nor phaclofen are potent specific GABAB receptor antagonists. PMID- 2556204 TI - Identification of a single alpha 1-adrenoceptor corresponding to the alpha 1A subtype in rat submaxillary gland. AB - 1. The alpha 1-adrenoceptors present in membranes of rat liver, cortex and submaxillary gland were labelled with [3H]-prazosin and the affinity of 15 ligands for these receptors was determined. 2. In saturation studies, [3H] prazosin bound with high affinity (Kd = 30-39 pM) to a single population of sites in all three preparations. 3. In competition studies using rat cortex, evidence for heterogeneity of the alpha 1-adrenoceptor binding sites was obtained. Displacement isotherms for amidephrine, benoxathian, oxymetazoline, phentolamine and WB 4101 were biphasic and were consistent with the presence of both alpha 1A- and alpha 1B-adrenoceptor subtypes as described by Morrow & Creese (1986) and Han et al. (1987). 4. The rat liver and submaxillary gland membrane preparations both possessed homogeneous populations of alpha 1-adrenoceptors. However, there were pharmacological differences between the receptors in these two preparations. Rat submaxillary gland alpha 1-adrenoceptors displayed high affinity for amidephrine, benoxathian, oxymetazoline, phentolamine and WB 4101 and therefore appeared to represent alpha 1A-adrenoceptors. Rat liver alpha 1-adrenoceptors possessed lower affinity for these ligands (6-65 fold) suggesting that these receptors were of the alpha 1B-subtype. 5. Spiperone exhibited 12.9 fold higher affinity for rat liver alpha 1B-adrenoceptors than for rat submaxillary gland alpha 1A adrenoceptor and may therefore represent the first alpha 1B-adrenoceptor selective ligand. PMID- 2556203 TI - GABA receptors on the somatic muscle cells of the parasitic nematode, Ascaris suum: stereoselectivity indicates similarity to a GABAA-type agonist recognition site. AB - 1. The gamma-aminobutyric acid (GABA) receptors on the somatic muscle cells of Ascaris, which mediate muscle cell hyperpolarization and relaxation, have been characterized by use of intracellular recording techniques. 2. These receptors are like mammalian GABAA-receptors in that the response is mediated by an increase conductance to chloride ions. The GABAA-mimetic, muscimol, has a relative potency of 0.40 +/- 0.02 (n = 3) compared to GABA. 3. The stereoselectivity of the GABA receptor on Ascaris is identical to that for the mammalian GABAA-receptor, as determined from the relative potency of three pairs of enantiomers of structural analogues of GABA. 4. The most potent agonist is (S) (+)-dihydromuscimol which is 7.53 +/- 0.98 (n = 5) times more potent than GABA. 5. The Ascaris GABA receptor is not significantly blocked, at concentrations below 100 microM by the potent, competitive GABAA-receptor antagonist, SR95531. 6. The Ascaris GABA receptor does not recognise agents that are known to block the GABA gated chloride channel in mammalian preparations such as t butylbicyclophosphorothionate (TBPS, 10 microM, n = 2) or the insecticide dieldrin (100 microM, n = 3). 7. GABAergic responses in Ascaris are not potentiated by pentobarbitone (100 microM, n = 3) or flurazepam (100 microM, n = 3). 8. The potencies of various GABA-mimetics in the Ascaris preparation have been compared with their potency at displacing GABAA-receptor binding in mammalian brain. Excluding the sulphonic acid derivatives of GABA, the correlation coefficient (r) between the potencies of compounds in the two systems is 0.74 (P less than 0.01). The significance of this correlation is discussed. 9. The pharmacology of the Ascaris GABA receptor is discussed in relation to other invertebrate systems and the mammalian subclassification of GABA receptors. PMID- 2556205 TI - Differences between the alpha 2-adrenoceptor in rat submaxillary gland and the alpha 2A-and alpha 2B-adrenoceptor subtypes. AB - 1. The alpha 2-adrenoceptors of rat submaxillary gland, labelled with [3H] rauwolscine, were characterized by use of a range of subtype selective ligands and were compared to rabbit spleen alpha 2A-adrenoceptors and rat kidney alpha 2B adrenoceptors. 2. In rat submaxillary gland, [3H]-rauwolscine labelled an apparently homogeneous population of binding sites with relatively low affinity (Kd = 11.65 nM) compared to the affinity in rat kidney (Kd = 2.18 nM) and rabbit spleen (Kd = 4.64 nM). 3. In competition studies using 16 ligands the alpha 2 adrenoceptors in rat submaxillary gland appeared to differ from both the alpha 2A adrenoceptor of rabbit spleen (r = 0.62) and also the alpha 2B-adrenoceptor of rat kidney (r = 0.28). 4. The affinity data obtained with benoxathian, imiloxan and WB 4101 indicated the presence of an alpha 2B-adrenoceptor in rat submaxillary gland. However, data for chlorpromaxine, oxymetazoline, spiroxatrine and xylometazoline indicated that submaxillary gland alpha 2-adrenoceptors were of the alpha 2A subtype. The affinity estimate for prazosin in rat submaxillary gland was intermediate between its affinity at the alpha 2A- and alpha 2B adrenoceptors while affinity estimates for idazoxan and phentolamine in rat submaxillary gland were greater than those obtained at either the alpha 2A- or alpha 2B-adrenoceptor. 5. These data indicate that rat submaxillary gland alpha 2 adrenoceptors differ from the alpha 2A- and alpha 2B-adrenoceptors found in rabbit spleen and rat kidney, respectively. PMID- 2556206 TI - Differential cardiovascular and respiratory responses to central administration of selective opioid agonists in conscious rabbits: correlation with receptor distribution. AB - 1. The effects of intracerebroventricular (i.c.v.) and intracisternal (i.c.) administration of a range of doses (0.01, 0.1 and 1.0 nmol kg-1) of specific mu- delta- and kappa-opioid agonists on cardiovascular and respiratory function and on plasma catecholamines have been studied in conscious rabbits. The distribution of mu- delta- and kappa-opioid receptors was localized in rabbit brain by in vitro autoradiography. 2. The mu-agonist [D-Ala2, MePhe4-Gly5-ol]enkephalin (DAGOL) given i.c.v. caused a large rise in plasma noradrenaline and adrenaline, hypertension accompanied by an initial bradycardia followed by tachycardia, respiratory depression and sedation. After i.c. administration there were similar changes in heart rate (HR) and respiration, but no significant changes in mean arterial pressure (MAP) or plasma catecholamines. 3. The delta-agonist [D Pen2.5]enkephalin (DPDPE) increased MAP and HR after both i.c.v. and i.c. administration, caused a small increase in noradrenaline but had no effect on adrenaline and did not alter respiration rate or blood gases. After i.c.v. DPDPE the rabbits became more alert and active. 4. The kappa-agonist U69593 given i.c.v. or i.c. had no effect on MAP or HR. After i.c.v. U69593, PaCO2 fell, but there were no other respiratory effects. The responses to dynorphin 1-13, an endogenous kappa-agonist, were similar to those of U69593. 5. The opioid antagonist naloxone (30 nmol kg-1) given intravenously (i.v.) blocked the effects of i.c.v. DAGOL (1 nmol kg-1). A 100 fold higher dose of i.v. naloxone (3 mumol kg-1) was required to abolish the effects of i.c.v. DPDPE (1 nmol kg-1). 6. Autoradiographic studies demonstrated a high density of mu- and delta-opioid receptors in hypothalamic sites. In the brainstem mu-receptors were demonstrated in the nucleus tractus solitarius (NTS) and delta-receptors in the dorsal motor nucleus of the vagus. kappa-Receptors were not detected in either the hypothalamus or brainstem. 7. These findings demonstrate that DAGOL increases sympatho-adrenal outflow, probably by stimulation of hypothalamic mu-receptors. The effects on HR are probably partly through a baroreflex and partly through an action of DAGOL on mu-receptors in the dorsal motor nucleus of the vagus. DPDPE probably acts on delta-receptors in the NTS to increase MAP and HR. Respiratory depression resulted from stimulation of mu-receptors in the brainstem with no evidence of delta- or kappa-receptors being involved. PMID- 2556208 TI - Extensor tendon rupture related to calcium pyrophosphate crystal deposition disease. PMID- 2556207 TI - Mediators of burn-induced neuromuscular changes in mice. AB - 1. Muscle paresis and aberrant pharmacological responses are two important pathophysiological changes that have been observed at the neuromuscular junction following thermal injury. By use of the mouse model of 20%, 30% and 50% total body surface area (BSA) burn, we examined the significance of intracellular mediators, adenosine 3':5'-cyclic monophosphate (cyclic AMP) and prostaglandin E2 (PGE2) in perturbing the physiological function of tension development and the pharmacological response to (+)-tubocurarine (+)-Tc at day 21 post-burn. 2. Cyclic AMP levels increased with the size of burn. The relationship between mean cyclic AMP levels and burn size was significant (R2 = 0.96, r = 0.98). Significant (P less than 0.05) reductions in tension development (g) were observed for the 30% and 50% BSA burn group compared to controls (30.3 +/- 8.3 and 34.1 +/- 5.9 vs 59.1 +/- 1.0, respectively). Tension alterations were associated with increased cyclic AMP levels; the relationship between increased cyclic AMP levels and tension decrease was significant (R2 = 0.82, r = 0.91). The dose of (+)-Tc required to inhibit twitch tension increased in proportion to burn size and was statistically significant in the 50% BSA burn group compared to controls (0.3320 +/- 0.09 vs 0.1093 +/- 0.11 mg kg-1, P less than 0.05). The alterations in the effective dose of (+)-Tc were significantly correlated to increases in cyclic AMP levels (R2 = 0.70, r = 0.83). Although PGE2 levels were elevated in the 30% and 50% burn groups, no relation was seen to either tension or (+)-Tc doses. 3. These studies, therefore, support the hypothesis that cyclic AMP plays a significant role in physiological and pharmacological responses in skeletal muscle following thermal injury. PMID- 2556210 TI - The use of place conditioning in studying the neuropharmacology of drug reinforcement. AB - The place conditioning paradigm has proven successful in identifying the neural mechanisms of drug reinforcement. Two classes of drugs, opiates and psychomotor stimulants, have received the most study, and in each case an important role for DA neurons of the mesolimbic system has been established. Moreover, both receptor subtypes, D1 and D2, appear to be involved. Despite this progress, the substrates of drug reward are not completely understood. First, a role for DA has not been established for all stimulants: DA receptor blockade failed to affect conditioned place preferences produced by the stimulants methylphenidate, nomifensine, or bupropion. Second, preliminary evidence suggests that intact serotonergic transmission is important in morphine place conditioning, but a similar consistent finding has not been observed with amphetamine place conditioning. Further study may reveal an interesting dissociation of serotonin's role in the rewarding effects of psychomotor stimulants and opiates. Finally, the role of the opiate receptor subtype kappa is not known; also, the significance of the several anatomical sites that support opiate place conditioning remains unclear. PMID- 2556209 TI - Progesterone enhances inhibitory responses of cerebellar Purkinje cells mediated by the GABAA receptor subtype. AB - Previous studies from this laboratory have indicated that the sex steroid progesterone (P) is able to augment inhibitory responses of cerebellar Purkinje cells to the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) in the urethane anesthetized adult female rat. Because both GABAA and GABAB receptors are localized in this CNS area, agonist/antagonists for these receptors were utilized in the present evaluation in order to delineate the specific receptor subtype involved in P-GABA interactions. Systemically administered P was found to augment GABA-mediated inhibition by an average of 80%. Concomitant application of the GABAA blocker, bicuculline, greatly reduced this modulatory effect of the steroid. In contrast, P administration had no effect on baclofen-induced inhibition. These data suggest that P effects on GABA physiology are mediated predominantly at the GABAA receptor. PMID- 2556211 TI - Blood-brain barrier permeability of human gliomas as determined by quantitation of cytoplasmic vesicles of the capillary endothelium and scintigraphic findings. AB - The number of cytoplasmic vesicles in the capillary endothelium was determined by ultrastructural morphometry and correlated with the uptake of technetium-99m pertechnetate used in brain scintigraphy. Ten gliomas were studied for uptake rates of 99mTc pertechnetate. Three gliomas from the different groups of uptake rates were quantitatively analyzed for cytoplasmic vesicle content. Capillaries of tumors without uptake had a low content of cytoplasmic vesicles, which was similar to that obtained in normal brain control. In tumors with low and moderate uptake rates, the cytoplasmic vesicles content increased significantly (p less than 0.05) by about 300% and 400%, respectively, as compared with that found in impermeable tumor and in normal brain. The correlation found between the cytoplasmic vesiculation of the endothelial cells in gliomas' capillaries and the uptake of 99mTc pertechnetate suggests that pinocytosis might be a factor in the uptake of the radionuclide. The present findings might be applicable to treatment with hydrophilic chemotherapeutic agents in moderate and highly permeable tumors. PMID- 2556214 TI - Fighting cancer in America. Achieving the "year 2000 goal". PMID- 2556212 TI - Evaluation of two management procedures for the control of maedi-visna. AB - Two control programs were evaluated for their efficiency in eradicating the maedi visna (M-V) virus from a single sheep flock. In both programs, the agar gel immunodiffusion test was used for the detection of M-V infected animals at regular intervals. In program 1, the test and remove program, ewes that were serologically positive for M-V were immediately removed along with their offspring. The prevalence of infected sheep decreased gradually and a seronegative flock was obtained after 30 months of monitoring. Program 2 entailed the removal of replacement ewe lambs at birth prior to the ingestion of colostrum. Maedi-visna antibodies have not been detected in this flock. These results show that under conditions similar to the industry norms, M-V can be expelled. Although the approach of program 1 is more practical for sheep producers, program 2 is more effective because of the earlier development of a M V seronegative flock. Because of the nature of the humoral response, a longer time period than four years is required to ensure that M-V has been completely eradicated from each flock. PMID- 2556213 TI - Protection of colostral antibodies against bovine leukemia virus infection in calves on a California dairy. AB - A three-year prospective study involving 244 calves was undertaken on a California dairy to evaluate the protective role of colostral antibodies against bovine leukemia virus (BLV) infection in calves. Calves were followed from birth to the time they left their individual hutch (TLIH), at about 90 days of age. The probability of being infected at TLIH and the daily risk of infection between birth and TLIH were modelled using the logistic and the Cox models, respectively. Calves with no detectable antibodies during the first week of life were up to 2.00 and 2.75 times more likely to be infected at TLIH compared to calves with low and high concentrations of antibodies during the first week of life, respectively (p = 0.01). When the daily risk was modelled, calves without antibodies at the estimated day of infection were up to 3.4 and 11.6 times more likely to become infected than calves with low and high concentrations of antibodies on that day, respectively (p less than 0.001). Results indicated that calfhood infection may be reduced by about 45% through the feeding of colostrum with BLV antibodies. Further reduction in infection may be possible by feeding calves milk powder, milk replacer, and/or milk from noninfected cows. Results also indicated that quantification of the effect of a time-dependent risk factor, such as colostral antibody concentration, might be affected if treated as a fixed factor. PMID- 2556215 TI - An in vitro system for study of effects of angiotensin I on cultured endothelial cells. AB - Since certain non-vascular angiotensin II (AII) receptors may be activated by angiotensin I (AI), and since sustained increase in AI levels accompanies chronic treatment with converting enzyme inhibitors (CEI) which block conversion of AI to AII, the question of whether AI has significant biological effects is of clinical relevance. We therefore sought to develop an in vitro culture system in which effects of angiotensin I, independently of its conversion to AII, could be studied in cloned aortic vascular endothelial cells (VEC). This was complicated by peptide degradation during the period of observation, both by angiotensin converting enzyme (ACE) on the surface of VEC and by angiotensinases in either the serum component of culture media or associated with the cell monolayer. Accordingly, we examined the half life of AI under relevant cell culture conditions, with and without confluent fetal bovine aortic endothelial cells (FBAEC). Factors assessed included (1) fetal calf serum: commercial source, concentration in culture media, effects of converting enzyme inhibitor (CEI: MK422) and/or heat inactivation (superimposed on the commercially performed process); and (2) effect of FBAEC in monolayer culture, with and without CEI. Results showed that (1) in the absence of cells, loss of AI in culture media, when present, was solely due to the presence of fetal calf serum (FCS) and showed a dose dependent response; (2) FCS from differing sources may vary dramatically in capacity for AI breakdown; and (3) serum related AI disappearance included a heat resistant ACE like component (inhibitable by CEI) and a heat sensitive/CEI resistant component dominant at concentrations of FCS exceeding 5%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556216 TI - Role of free radicals and neutrophils in canine myocardial reperfusion injury: myocardial salvage by a novel free radical scavenger, 2-octadecylascorbic acid. AB - To define the role of oxygen free radicals and neutrophil involvement in evolving myocardial reperfusion injury, we evaluated the effect of 2-octadecylascorbic acid (CV-3611), a novel free radical scavenger, on neutrophil function and the extent of myocardial damage resulting from 90 min of ischaemia followed by 5 h of reperfusion in an experimental model of myocardial infarction. Dogs were randomly assigned to receive CV-3611 (5 mg.kg-1.[5 min]-1, intravenously) just before the onset of reperfusion. Infarct size, as a percent of area at risk, was reduced by 60% in CV-3611 treated group as compared with control, at 16.7(SEM 3.1)% v 41.5(4.5)%, p less than 0.01. Administration of CV-3611 markedly reduced function of neutrophils isolated from peripheral circulation during reperfusion ex vivo as estimated by free radical generation (ferricytochrome c reduction and luminol enhanced chemiluminescence), chemotactic activity, and aggregation induced by A23187. Under these conditions, the enhancement in neutrophil infiltration and free radical generation (luminol enhanced chemiluminescence) in myocardium within area at risk, especially in the border zone between viable and irreversible injured myocardium, was markedly reduced. Haemodynamic profiles were similar between control and CV-3611 treated group. These results suggest that activated neutrophils, especially their generation of oxygen free radicals, contribute to reperfusion induced myocardial injury. PMID- 2556217 TI - Regulation of synaptic transmission in the central nervous system: long-term potentiation. PMID- 2556218 TI - Molecular cloning and characterization of a human DNA binding factor that represses transcription. AB - Several transcription factors interact with GC-rich sequences and positively regulate both housekeeping genes and cellular oncogenes. We have cloned a human cDNA that encodes a factor that binds to the GC-rich sequences present in the epidermal growth factor receptor (EGFR), beta-actin, and calcium-dependent protease (CANP) promoters. This cDNA encodes a 91 kd protein with an extremely basic region at its amino terminus. Deletion analyses with bacterially expressed proteins containing fragments of this factor indicate that this basic region of the protein functions as the DNA binding domain. Expression of this factor in CV1 cells shows that it represses expression originating from both the EGFR and beta actin promoters as well as chimeric promoters containing the CANP gene. It also represses transcription in cell-free extracts. These results suggest that positive and negative factors may interact with the same control element to account for the diversity of transcriptional regulation. PMID- 2556219 TI - Progressive paresthesias after cessation of therapy with very high-dose cisplatin. AB - Control of cisplatin-induced nephrotoxicity and nausea/vomiting has enabled the development of very high-dose cisplatin regimens (monthly total dose, 200 mg/m2). Neurotoxicity is now recognized to be the dose-limiting toxicity of these regimens. However, during a pilot study involving 5 mg/m2 vinblastine and 100 mg/m2 cisplatin given every 28 days on days 1 and 8 for the treatment of advanced non-small-cell lung cancer, we noted a high incidence of progressive peripheral neuropathy, which continued for several months after the discontinuation of cisplatin chemotherapy. Of the six patients treated, four received at least three cycles of therapy (median total cisplatin dose, 685 mg/m2; range, 500-725 mg/m2). All four patients developed a progressive peripheral neuropathy, with a worsening of toxicity by 1-3 grades over the 2-3 months after cisplatin discontinuation. One patient progressed from grade I (mild paresthesia) to grade IV (inability to ambulate) over a period of 3 months after the discontinuation of therapy. Stricter rules for early dose de-escalation and discontinuation may be required for very high-dose cisplatin regimens. Delayed progressive neuropathy should be recognized as a possible late complication of this form of therapy. PMID- 2556221 TI - [Inhibitory effect of electroacupuncture on cAMP induced ECOG epileptiform waves]. AB - In conscious rabbits, injection of cAMP (100 micrograms in 100 microliters, icv) elicited high-amplitude and high-frequency epileptiform seizure pattern of ECoG. The epileptic waves were inhibited by electro-acupuncturing bilateral "Zusanli" Points. During and after the cessation of electro-acupuncture the frequency, amplitude and duration of epileptiform discharges decreased significantly, the seizure waves even disappeared completely. The frequency and amplitude of epileptic waves showed significant difference in comparison with those of the non electroacupuncture group (p less than 0.001 and p less than 0.01 respectively), the duration of epileptic waves shortened by 58.18-66.88%. The results suggested that electroacupuncture has antiepileptic action on c-AMP induced experimental epileptic seizure. PMID- 2556220 TI - Enhancement of the invasive ability of a transformed human bronchial epithelial cell line by 12-O-tetradecanoyl-phorbol-13-acetate and diacylglycerol. AB - The effect of the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA) was studied using an immortalized human bronchial epithelial cell line, BEAS-2B, both in vivo and in vitro. The in vivo model consisted of tracheas reconstituted with an epithelium of BEAS-2B cells xenotransplanted into athymic nude mice. Intraluminal TPA treatment caused increased BEAS-2B cell proliferation and downgrowth into the tracheal stroma. In an in vitro invasion assay, TPA enhanced the invasive capacity of BEAS-2B cells 20- to 25-fold. A similar result was observed with diacylglycerol (DAG), an endogenous activator of protein kinase C, and the effects of TPA and DAG were abolished by simultaneous treatment with H-7, a protein kinase C inhibitor. TPA induced type IV collagenolysis, and this effect also was prevented by H-7. These data are consistent with the hypothesis that TPA causes these cells to become invasive by inducing collagenase activity and that this effect is mediated via protein kinase C. PMID- 2556222 TI - Effect of ingestion of hemoproteins on fecal excretion of hemes and porphyrins. AB - Stools from asymptomatic volunteers on diets containing red meat, whole blood, or high fiber were analyzed for their content of hemes and dicarboxylic (heme derived) porphyrins by the "HemoQuant" assay, the "Hemoccult" test, and "high performance" liquid chromatography (HPLC). In 49 subjects, ingestion of red meat increased HemoQuant-determined combined fecal heme plus dicarboxylic porphyrins by an average 375%; the contribution of heme-derived porphyrins to total fecal porphyrins increased from 37% to 78%. Of subjects on a red-meat diet, 27% passed stools with a porphyrin content suggestive of a porphyria, compared with only 4% on a red-meat-free diet. These increases were due largely to protoporphyrin and its derivatives pemptoporphyrin and deuteroporphyrin, all of which were present in feces as iron-free porphyrins and iron-ligated (heme) forms. Ingestion of blood had an effect similar to that of red meat, but ingestion of fiber had no effect. These effects of dietary and endogenous hemoproteins must be considered when such methods are used to test feces for occult blood or to test for excess fecal porphyrins as an indicator of a porphyria. PMID- 2556224 TI - Effects of intranasal calcitonin administration on pituitary GH response to hGHRH (1-29)NH2 in normal adult subjects. AB - Studies in man demonstrated that intramuscular salmon (s) calcitonin (CT) administration blunted pituitary GH response to hypothalamic stimuli such as arginine infusion and insulin-induced hypoglycaemia. However, the mechanisms underlying this inhibiting action of CT are still unclear. The aim of our study was to investigate the effects of intranasal (i.n.) and i.m. sCT administration on GH secretion elicited by direct pituitary stimulation in man with human GH releasing hormone (GHRH) (1-29)NH2. Seven healthy non-obese volunteers (five men, two women; mean age +/- SDM 25 +/- 2) underwent a bolus intravenous injection of GHRH, 100 micrograms, alone or associated with sCT, administered either i.n., 300 IU, or i.m., 100 IU. Our data demonstrate a significant decrease in GH secretion after GHRH when either i.n. or i.m. sCT is administered. GH peak (mean +/- SDM); GHRH alone 22.9 +/- 2.5 vs GHRH plus i.n. sCT, 8.9 +/- 1.5 micrograms/l, P less than 0.001; or vs GHRH plus i.m. sCT 12.3 +/- 2.5 micrograms/l, P less than 0.001. Area under the curve of GH secretion: GHRH alone 1211 +/- 196 vs GHRH plus i.n. sCT 551 +/- 116 micrograms 120 min/l. P less than 0.001; or vs GHRH plus i.m. sCT 700 +/- 167 micrograms 120 min/l, P less than 0.001. We conclude that sCT is able to inhibit GHRH-stimulated GH secretion in man. PMID- 2556223 TI - Monoamine oxidase (MAO) activity and its forms in normal and pathological human thyroid tissues. AB - Monoamine oxidase (MAO) activity and its forms were measured in normal human thyroid glands (39 samples from necropsy and four from perinodular tissues) and in pathological thyroid glands (13 multinodular goitres, three carcinomas, seven chronic thyroiditis, five Graves' disease and seven follicular, seven foetal and five embryonal adenomas). MAO activity in carcinomas was lower than in normal perinodular thyroid control tissue; in Graves' disease, foetal and embryonal adenomas it was significantly higher than in the normal control. The use of clorgyline (a selective MAO A inhibitor) and deprenyl (selective MAO B inhibitor) allowed estimation of the relative proportion of MAO forms. A high proportion of MAO A (more than 90%) was observed, both in normal and in pathological thyroid tissues. Changes in total thyroid MAO activity, but not in its forms, were found in different pathologies of the thyroid gland. PMID- 2556225 TI - The effect of in vitro exposure to histamine on mononuclear leucocyte phosphodiesterase activity in atopic dermatitis. AB - Increased cyclic AMP (cAMP) phosphodiesterase activity (PDE) is present in the peripheral blood mononuclear leucocytes (MNL) of patients suffering from atopic dermatitis. It is unknown whether this is a primary abnormality or whether it is a consequence of MNL exposure to inflammatory mediators. In this study we have compared the responses of MNL PDE from untreated atopic dermatitis patients, patients on prolonged therapy with topical fluorinated steroids, and normal controls following exposure in vitro to low concentrations of histamine. We have confirmed that PDE activity is elevated in the MNL of patients suffering from atopic dermatitis who have used emollients only for 1 week. However, patients on therapy with topical fluorinated steroids have MNL PDE activity similar to normal controls in spite of the persistence of some active eczema. Exposure to histamine in vitro results in a similar increase of MNL PDE in untreated patients and normal controls. However, treated patients show no elevation of MNL PDE activity following in vitro histamine exposure. The results suggest that the normal MNL PDE values seen in patients treated with topical fluorinated steroids may not be the consequence of reduced cutaneous inflammation, but the result of a direct action of topical steroids on MNL. This action abrogates any in vitro response to histamine and presumably acts in a similar fashion in vivo. PMID- 2556226 TI - An electron microscopic study of the epidermal infiltrate in recurrent herpes simplex. AB - Host immunity has been suspected of playing a role in recurrent herpes simplex. In this preliminary ultrastructural study of two patients with acute herpetic eruption, it was noted that the keratinocytes exhibiting the most severe damage are those adjacent to large granular lymphocytes. In contrast, many keratinocytes filled with viral particles of herpes simplex show little or no signs of keratinocyte damage. These observations suggest that in recurrent herpes simplex the epidermal damage may be due, at least in part, to cell-mediated host immunity as well as to the viral infection. PMID- 2556227 TI - Cutaneous ulcers due to Candida albicans in an immunocompromised patient- response to therapy with itraconazole. AB - The patient, a 68-year-old female, with chronic anaemia and a previous history of carcinoma of the breast treated surgically, had been receiving therapy with methyl prednisone, in addition to gold, for pemphigus vulgaris. She developed deep cutaneous ulcers on the lower leg. All had well-defined edges and were covered with purulent and serosanguinous exudates. On histopathology the ulcers were deeply infiltrated with yeasts and mycelium and Candida albicans was isolated on culture. There was no evidence of systemic candidosis. Complete healing was obtained using itraconazole in a dose of 200 mg daily for 45 days: treatment with prednisone was continued throughout. The response to antifungal therapy alone suggests that Candida was largely, if not wholly responsible, for this unusual clinical condition. PMID- 2556228 TI - Modulation of the iodination reaction in normal human neutrophils and in whole blood by penicillamine, congeners and intracellular enzyme catalase and superoxide dismutase. AB - We have demonstrated that penicillamine (PSH) has the capacity to effect phagocytic cells by its interaction with the myeloperoxidase-halide system (MPOHS). We have undertaken studies at the cellular level, measuring the activity of the MPOHS through quantitation of I125 uptake (free and bound), known as the iodination reaction (i.e., in isolated polymorphonuclear leukocytes (PMN) and in whole blood). In contrast to other studies investigating the effects of PSH on isolated myeloperoxidase (MPO), we have shown that PSH scavenges the H2O2 produced by phagocytic cells, thereby reducing the availability of H2O2 for conversion to HOI125 by myeloperoxidase. This was observed as a reduction in the level of iodination. This finding is supported by our having obtained similar results in PMN and whole blood with catalase (C), glutathione (GSH) and N acetylcysteine (NAC) but not with penicillamine disulphide (PSSP) or superoxide dismutase. Cu2+ (8 microM) when incubated with PSH reduced the level of inhibition of the iodination reaction by the oxidation of PSH to PSSP, illustrating the importance of the free sulphydryl group for this action. Incubation of PMN or whole blood for 0 to 2 hours with PSH, with a subsequent washing of PMN prior to stimulation, showed that PSH (free) requires to be present during stimulation of phagocytic cells to have this effect on the iodination reaction. Superoxide dismutase (SOD) produced increases in the iodination reaction in stimulated PMN by increasing the availability of H2O2. In conclusion, PSH inhibits the myeloperoxidase-halide system at a cellular level by scavenging H2O2 rather than by oxidation of the myeloperoxidase enzyme. This was observed at clinically relevant concentrations of PSH. PMID- 2556229 TI - Rheumatoid arthritis associated with osteopoikilosis: a case report. PMID- 2556230 TI - Alterations in arachidonic acid metabolism and chemotactic response in polymorphonuclear leukocytes from patients with rheumatoid arthritis. AB - This study examines the interrelationship between arachidonic acid (AA) metabolism and chemotactic potential of human peripheral blood neutrophils from both normal donors and patients with rheumatoid arthritis (RA). We demonstrate that there is a significant inverse relationship between the neutrophil's metabolic capability to produce [3H]LTB4 in response to ionophore A23187 stimulation versus the cell's chemotactic potential to optimal concentrations (i.e., 20 nM) of the chemotactic peptide f-Met-Leu-Phe as determined by leading front (r = 0.567, p = 0.009), mean depth (r = 0.458, p = 0.042), and population ratio (r = 0.471, p = 0.036) analyses. Subsequent Percoll density gradient studies revealed several RA neutrophil subpopulations exhibiting significantly enhanced (p less than 0.05) [3H] AA metabolism over corresponding density normal cells. Based on these results, we speculate that RA peripheral blood neutrophils have been "processed" to become less chemotactically-responsive yet more metabolically-active cells, and that the increased ability to produce LTB4 through both increased phospholipase A2 and lipoxygenase activities are part of the increased metabolic capabilities of the cell. PMID- 2556232 TI - A comparison of two APTT reagents which use silica activators. AB - The Ortho activated partial thromboplastin time (APTT) reagent, Thrombosil 1 (TS), was compared to the General Diagnostics automated APTT reagent (GD). TS produced more precise results over a 38-day period of testing a normal control plasma, indicating that the upper limit of the normal range could be more precisely set with TS. This normal range was better represented if the normal values with both reagents were logarithmically transformed before calculating the mean +/- 2 SD. TS was more sensitive to plasma which had been heparinized in vitro. This was also demonstrated in vivo by the testing of 100 plasmas from heparinized patients. On testing of in-vitro dilutions of normal plasma with factor-deficient plasmas, TS was more sensitive to decreasing levels of factors VIII, IX and XI but less sensitive to decreasing factor XII. This was demonstrable in vivo in 71% of cases with plasmas from factor-deficient patients. GD was more sensitive to the lupus anticoagulant in most cases. PMID- 2556231 TI - Peripheral nervous system involvement in essential cryoglobulinemia and nephropathy. AB - The clinical and neurophysiological features of 23 patients affected by essential cryoglobulinemia (EC) have been studied. It was possible to perform sural nerve biopsy in 3 cases. Six patients were found to be affected by a peripheral neuropathy, according to the WHO criteria, while in 8 other patients clinical and neurophysiological signs of a milder peripheral nervous system (PNS) involvement were evident. The incidence of PNS involvement seems to be high (60.9%). Neurophysiological and histological studies were indicative of a mainly axonal damage. PMID- 2556233 TI - Intravenous immunoglobulin treatment of Epstein-Barr virus-related idiopathic thrombocytopenic purpura. PMID- 2556234 TI - Enzymes in sarcoidosis. Angiotensin-converting-enzyme (ACE). AB - Measurement of serum angiotensin-converting-enzyme (SACE) is the most useful test for diagnosing and monitoring disease activity in sarcoidosis. Because elevated levels of SACE are not specific for sarcoidosis, other conditions in which SACE levels are also elevated are examined. PMID- 2556235 TI - Evaluation of a HSV specific monoclonal antibody reagent for laboratory diagnosis of herpes simplex virus infection. AB - A new FITC-conjugated HSV specific monoclonal antibody reagent (Syva Co., Palo Alto, Ca) was evaluated for the confirmation of HSV clinical isolates. The reagent was also compared to type-specific monoclonal antibodies for the pre-CPE detection of HSV from clinical specimens in centrifugation culture and by direct examination of specimens smears by direct immunofluorescent antibody staining (DFA). HSV was isolated from 75 of 232 specimens (32%). All 75 isolates were confirmed with both the type-specific antibodies and the HSV-specific reagent. In centrifugation culture HSV was detected in 36 of 105 (34%) specimens. The HSV specific reagent detected all 36 specimens that were positive with the type specific reagents. HSV infection was diagnosed by DFA in 31 of 50 (62%) specimen smears. The HSV-specific reagent detected all 31 positive specimens. This reagent confirmed and detected all HSV positive specimens that were positive by the type specific monoclonal antibody reagents. The reagent contains monoclonal antibodies specific for both HSV1 and HSV2 in a single mixture, which produces a highly sensitive HSV FA staining reagent. PMID- 2556236 TI - Herpesvirus simiae contamination of primary rhesus monkey kidney cell cultures. CDC recommendations to minimize risks to laboratory personnel. PMID- 2556237 TI - Rapid culture confirmation of adenovirus isolates using the adenoclone enzyme immunoassay (EIA) test kit. AB - The adenoclone enzyme immunoassay (EIA) test kit (Cambridge BioScience Corp., Worcester, MA) was evaluated using 43 adenovirus and 53 nonadenovirus (i.e., polio-, echo-, coxsackievirus types A,B, parainfluenza virus, and herpes simplex virus) isolates obtained during routine clinical diagnostic screening procedures. Forty-two of 43 confirmed adenovirus isolates were reactive upon testing by the adenoclone test kit. None of the 53 nonadenovirus isolates, including seven difficult-to-cultivate group A coxsackievirus isolates, cross-reacted with the EIA kit. The adenoclone EIA kit is an appropriate supplement to the regimen of clinical diagnostic procedures for the culture confirmation of adenovirus. PMID- 2556238 TI - The in vitro effect of sulbactam on polymorphonuclear leukocyte function. AB - The effects of sulbactam on polymorphonuclear leukocyte (PMN) chemotaxis, respiratory burst, and microbicidal activity against Staphylococcus aureus were evaluated in vitro. PMNs from normal adult volunteers were incubated with 0.5, 2, 5, 10, and 20 micrograms/ml of sulbactam for 30 min each. At concentrations of 5 and 10 micrograms/ml, sulbactam was found to enhance PMN bactericidal activity. No inhibitory effects on PMN function were noted at the concentrations of sulbactam that were tested. In addition, the in vitro inhibitory effect of ampicillin on PMN chemiluminescence was partially abrogated by the presence of sulbactam. These findings suggest that the microbicidal phagocytic response of human PMNs against S. aureus may be improved in vitro using concentrations of sulbactam that may be readily achieved with therapeutic doses of sulbactam/ampicillin. PMID- 2556240 TI - Efficacy of fluconazole in cryptococcal meningitis. AB - In this clinical trial, oral fluconazole was used to treat cryptococcal meningitis in 32 patients with Acquired Immune Deficiency Syndrome (AIDS). In 11 patients who received 200 to 400 mg/day of fluconazole as primary therapy, a favorable clinical response was obtained in 67% of all evaluable patients. A negative cerebrospinal fluid (CSF) culture was also reported for 86% of these cases. Fluconazole was used as second-line therapy in an additional 15 patients who were not responsive to therapy with amphotericin B or amphotericin B combined with flucytosine. Positive clinical and mycologic responses were then obtained in more than 60% of these cases. Following successful treatment with fluconazole as either the primary or secondary antifungal agent, 26 patients were evaluated during maintenance therapy with 100 to 200 mg daily of fluconazole to prevent recurrence of disease. The relapse rate was 3.2 cases of cryptococcal meningitis per 1000 patient weeks, with a mean duration of 22 weeks for maintenance therapy. An additional six patients who were also treated with either amphotericin B alone or in combination with flucytosine but were asymptomatic or CSF culture negative when treatment with fluconazole was initiated were evaluated for the safety and efficacy of maintenance therapy. Thus, treatment of fluconazole appears to be efficacious as well as safe. The incidence of superimposed infections associated in these AIDS patients make it difficult to accurately assess any adverse effects. PMID- 2556239 TI - Review of fluconazole: a new triazole antifungal agent. AB - Fluconazole is a new triazole antifungal agent with unique pharmacokinetic properties. It can be administered orally or parenterally and achieves rapid distribution by either route, and its absorption is not affected by the presence of food. It has a plasma half-life of approximately 25-30 hr and approximately 70% of dose is excreted in the urine unchanged. There is linearity of fluconazole plasma concentrations over the dose range and the elimination rate is independent of dose. No effect has been seen on basal or ACTH-stimulated cortisol or on testosterone, estrogen, progesterone, or other steroid hormones, and there is no interaction with an oral contraceptive. No interaction with concomitantly administered cyclosporine has been documented, and there are no clinically significant differences in absorption when fluconazole is given in the presence or absence of cimetidine or food. Patients who are concomitantly receiving coumarin anticoagulants should be monitored because there is an interaction between fluconazole and such anticoagulants. Patients taking oral hypoglycemics and fluconazole should be monitored, because fluconazole has been shown to inhibit the metabolism of tolbutamide. Fluconazole has been successfully used to treat a variety of fungal infections in a variety of contexts including vaginal candidiasis; oropharyngeal candidiasis in immunocompromised patients, those with malignancies, transplant recipients, and patients with systemic sclerosis; patients with cryptococcal meningitis; and patients with fungal infections who were also treated with chemotherapy or radiation therapy. In the treatment of all of these infections with doses ranging from 50 mg to 400 mg a day of fluconazole, there has been a very low incidence of side effects (9.3%) reported, and only 1.1% of all patients were withdrawn from therapy. PMID- 2556241 TI - A clinical study of fluconazole for the treatment of deep mycoses. AB - A multicenter clinical study of fluconazole was conducted at 41 hospital sites in Japan. Fluconazole was administered orally or intravenously at daily doses of 50 to 400 mg to 199 patients with deep-seated mycoses. Clinical efficacy was evaluable in 125 of these patients. Most cases were complicated with serious underlying diseases such as cancer, leukemia, or AIDS. Clinical cures were achieved in 56 (87.5%) of 64 cases of candidiasis, in 11 (68.8%) of 16 cases of cryptococcosis, in 19 (44.2%) of 43 cases of aspergillosis, and in one case each (100%) of mucormycosis and fungemia due to an unspecified yeast. Eradication rates of causative fungi were 87.9% in Candida spp., 62.5% in Cryptococcus neoformans, and 52.2% in Aspergillus spp. Side effects were observed in 13 cases, with an incidence rate of 6.5%. In most cases fluconazole was well tolerated. Changes in laboratory test values due to the drug were reported in 35 patients with an incidence rate of 17.6%. The changes were minor and transient; primarily increases in liver enzyme. Fluconazole is a useful antifungal agent for the treatment of systemic deep-seated mycoses. PMID- 2556242 TI - Difference in thiamine metabolism between extensor digitorum longus and soleus muscles. AB - 1. Thiamine diphosphate level was higher in soleus muscle than in extensor digitorum longus muscle in various animals, whereas thiamine triphosphate level was less in the former muscle than in the latter except for mouse. 2. 2 Oxoglutarate dehydrogenase, transketolase and thiamine pyrophosphokinase activities were higher in soleus muscle than in extensor digitorum longus in rat and guinea pig. 3. The differences between rat two muscle phenotypes in thiamine diphosphate, but not thiamine triphosphate, level and the thiamine-related enzyme activities disappeared after denervation. 4. Tenotomy had little effect on thiamine phosphate levels and the thiamine-related enzyme activities in rat skeletal muscles. PMID- 2556243 TI - Septic plasma suppresses superoxide anion synthesis by normal homologous polymorphonuclear leukocytes. AB - In vitro exposure of normal O,Rh- PMN to plasma obtained from patients with septic shock results in inhibition of formyl-methionyl-leucyl-phenylalanine stimulated superoxide anion (O2.-) production by 40%. Although all reaction velocities and extent of reaction at 5 min were suppressed, neither lag time preceding O2.- production nor duration of initial velocity linearity was affected. No such inhibition was noted when plasma from healthy controls or nonseptic critically ill patients was utilized in the reaction. These results demonstrate that neutrophils are not only a cause, but also a target of the septic shock host inflammatory response. PMID- 2556244 TI - Phagocyte-pathogen in the infected host. AB - Phagocytosis is a major mechanism of defense against bacterial infections. The ingestion of different microorganisms by blood granulocytes or monocytes may involve a variety of cell membrane recognition structures (e.g., immunoglobulin or complement receptors, lectin-like structures or other nonspecific binding sites). It is of interest to know which mechanism plays a prominent role in the management of a particular type of infection. Forty-three pathogenic bacterial suspensions were obtained from patients under mechanical ventilation at the onset of nosocomial lower respiratory tract infections. They were coincubated with blood granulocytes from the same or other patients in the presence or absence of the corresponding serum. Phagocytosis and the oxidative burst were then assayed. We conclude the following: a) Substantial phagocytosis was found under serum-free conditions and the patients' sera did not dramatically enhance bacterial uptake during the first days after the onset of clinical symptoms. b) The phagocytes from an infected patient did not display any peculiar inability to bind to the bacteria that grew successfully in this subject. Hence, the occurrence of a particular infection might be dependent on a defect of intracellular killing of ingested pathogens or on the conditions of infection development. PMID- 2556246 TI - Method for the detection of tissue metabolite (H2(17)O) in brain by proton magnetic resonance imaging. PMID- 2556245 TI - Brain pH effects of NaHCO3 and Carbicarb in lactic acidosis. AB - The effects of iv sodium bicarbonate (NaHCO3) and Carbicarb, an experimental buffer, were compared in a rat model of lactic acidosis induced by controlled hemorrhage and asphyxia. Although both NaHCO3 and Carbicarb were effective at alkalinizing the arterial blood in this model, NaHCO3 administration resulted in a rise in PaCO2 where Carbicarb did not (+9 +/- 2 vs. +2 +/- 2 torr at 2 min after infusion, p less than .01). Moreover, NaHCO3 resulted in a small decrease in intracellular brain pH as measured with P-31 nuclear magnetic resonance where Carbicarb afforded intracellular brain alkalinization (-0.03 +/- 0.01 vs. +0.08 +/- 0.02 pH units at 2 min, p less than .01). If these data are confirmed clinically, Carbicarb may offer advantages over NaHCO3 under conditions of fixed or limited ventilation. PMID- 2556247 TI - [Pulmonary metastases of malignant tumors of the oral and maxillofacial region. Analysis of 70 cases]. AB - Records of 685 cases of malignant tumors in the oral and maxillofacial regions were collected. Pulmonary metastases occurred in 70 of these cases. The metastatic rate was 10.2%. Roentgenographic appearances of pulmonary metastases were divided into 3 types: (1) Multiple ball-like lesion (78%); (2) Single ball like lesion (14%); (3) Irregular lesion (8%). When metastasis was single, diagnosis was made according to the location of the metastasis and comparison between and old recent roentgenograms. Pulmonary metastasis was more frequent in tumors of the submandibular gland (34.5%), the base of the tongue (33.3%), and the sublingual gland (27.3%). Adenoid cystic carcinoma (47.8%) and adenocarcinoma (26.5%) might readily develop pulmonary metastasis. Pulmonary metastasis was rare in squamous cell carcinoma (2.5%). The metastatic rates were 3.9%, 5.4%, 6.5%, and 7.1% in stage I, II, III and IV, respectively. The period between the occurrence of pulmonary metastasis and initial treatment was from 1 month to 8 years and 5 months, with a mean of 33.8 months. The period between the occurrence of pulmonary metastasis and death of the patient was from 1 month to 8 years and 8 months, with a mean of 21.4 months. PMID- 2556248 TI - [Diagnostic significance of the determination of serum alpha-L-fucosidase in primary hepatocellular carcinoma]. AB - The results of determination of serum alpha-L-fucosidase (AFU) in healthy individuals, in patients with various liver diseases and non-liver malignant diseases were reported. In primary hepatic carcinoma the level of serum AFU was significantly higher than that in various other diseases and healthy persons (P less than 0.001). The serum AFU level was independent of AFP level (x = 1.24, P greater than 0.25). These data suggest that AFU may be a useful marker for the diagnosis of primary hepato cellular carcinoam with negative AFP. PMID- 2556249 TI - [Immunologic changes in serum CuZn-containing superoxide dismutase in patients with lung cancer]. AB - Serum immunologic changed CuZn-containing superoxide dismutase (CuZn-SOD) level was determined in 45 patients with lung cancer and the results were compared with those in 22 normal persons and 19 cases with benign pulmonary diseases. The results were as follows: x +/- Sx of CuZn-SOD in the lung cancer group was 16.11 +/- 0.78 ng/ml, that in the benign pulmonary disease group was 9.51 +/- 0.61 ng/ml and that in the normal adults group was 7.22 +/- 0.79 ng/ml. The mean of the activity of serum immunologic changed CuZn-SOD in patients with lung cancer was significantly high than that in the other two groups. (P less than 0.001). If a serum immunologic changed CuZn-SOD level of 11.98 ng/ml and over is considered as Positive, the positive rate will be 80%, 11%, and 4.5% in the three groups respectively. In the light of the above findings, we suggest that determination of the activity of serum immunologic changed CuZn-SOD is helpful in the diagnosis of lung cancer. PMID- 2556250 TI - [Sensorimotor and autonomic neuropathy associated with systemic lupus erythematosus]. AB - In this article, one described 7 patients with systemic lupus erythematosus (SLE) and the pathological findings of their sural nerves. In 4 of the 7 cases the criteria for diagnosis of SLE were satisfied and in 3 cases there was serological evidence of an undifferentiated connective tissue disease, most likely to be SLE. The peripheral neuropathy was of a chronic sensorimotor type with predominantly sensory features and gradual onset. In 2 cases the presentation was asymmetrical. One patient had autonomic dysfunction. There was ischemic neuropathy in 2 cases, segmental demyelination in 2 cases, and axonal degeneration with demyelination in 3 cases. In 6 cases there was increased expression of Ia antigen within the nerve fascicle, perineurium and endothelial cells. PMID- 2556251 TI - [Advances in clinical studies and experimental research on toxic neuropathy]. PMID- 2556252 TI - Anal cancer and human papillomaviruses. AB - Epidemiologic and clinical evidence has suggested a possible association between anal cancer and human papillomavirus (HPV) types that are known to be associated with cervical and other genital cancers. Using Southern blot and dot blot analysis, the authors examined 45 primary anal malignancies for HPV DNA types 6, 11, 16, and 18. HVP 16, DNA was detected in 23 of 41 (56 percent) anal squamous cell carcinomas (SCC) and in the lymph-node metastases of two of these tumors. In addition, HPV 18 DNA was detected in 2/41 (5 percent) anal SCCs. Anal SCC contained no detectable HPV 6 or 11 DNA. The remaining four primary anal malignancies were not squamous carcinomas and did not contain any detectable HPV DNA. Nonmalignant anal epithelium and malignant rectal mucosa obtained from surgical patients undergoing hemorrhoidectomy and abdominoperineal excision of the rectum did not contain any detectable HPV DNA. HPV 16 DNA in anal cancer was predominantly integrated into the host cell DNA. In situ hybridization was used to demonstrate that HPV 16 DNA in anal SCC tissues is confined to the nuclei of carcinoma cells. The results of this investigation closely parallel similar studies of cervical cancer and lend support to the concept of the involvement of HPV 16 and 18 in the development of anal and genital squamous-cell carcinoma. PMID- 2556254 TI - HPV 16-positive bowenoid papulosis and squamous-cell carcinoma of the anus in an HIV-positive man. AB - A homosexual man in stage IV of HIV infection, who suffers from HPV 16-positive bowenoid papulosis of the anal region, is described. In one area the patient developed an HPV 16-positive squamous-cell carcinoma of the anus. Bowenoid papulosis represents a squamous-cell carcinoma in situ, and usually follows a benign clinical course. The possibility exists that immunocompromised individuals are at higher risk to develop cancer on the basis of bowenoid papulosis. PMID- 2556253 TI - Ileal adenomas in postcolectomy patients with familial adenomatosis coli/Gardner's syndrome. Incidence and endoscopic appearance. AB - Endoscopy and biopsy of the terminal ileum were performed in 18 patients with familial adenomatosis coli/Gardner's syndrome. All had undergone total colectomy with ileoproctostomy 7 to 249 months (average, 79.7 months) before the study. In all of these patients, endoscopic studies revealed multiple or innumerable small (less than 4 mm in diameter) polypoid lesions, all recognizable as whitish, sessile elevations. Histologic findings of the biopsy specimens from the polypoid lesions showed tubular adenoma, with or without lymphoid hyperplasia, in nine (50 percent), but only lymphoid hyperplasia in the other nine patients. Colonic metaplasia was present in the adjacent ileal mucosa in 3 patients with ileal adenomas. The incidence (83 percent) of ileal adenomas detected 113 to 249 months after colectomy was higher than that (33 percent) found 7 to 90 months after surgery. In view of these results, endoscopy and biopsy of the terminal ileum, as well as the retained rectum, should be done periodically for postcolectomy patients with this disease. PMID- 2556255 TI - Small cell lung cancer. AB - Lung cancer is the most common cause of cancer death in the United States, with approximately 135,000 men and women dying each year. While much has been learned about the etiologic risk factors, less progress has been made in therapy. Five year survival rates remain at less than 10%. However, there has been some progress in the therapy of one histological subtype of lung cancer, small cell lung cancer. Totaling around 20% of lung cancer cases, small cell lung cancer is distinct from the other histologic subtypes in its biologic behavior and responsiveness to therapy. In the 1960s, the median survival for patients with small cell lung cancer was approximately 3 months. With combination chemotherapy and radiotherapy median survivals now range from 1 to 2 years, and there is evidence for a curative potential since approximately 10% of patients who initially present with limited disease survive greater than 2 years. The unique clinical aspects of this histological subtype potentially relate to its underlying cell of origin. This behavior is reflected in the numerous paraneoplastic syndromes that frequently accompany small cell lung cancer. Its propensity for early dissemination have made staging the extent of disease an important part of the clinical evaluation. Since small cell lung cancer is sensitive to both chemotherapy and radiation therapy, there have been multiple clinical trials evaluating drug/radiotherapy combinations. This article will briefly describe the unique aspects of small cell lung cancer as opposed to other histological subtypes of lung cancer and give an overview of the current clinical approach and treatment of this disease. PMID- 2556257 TI - [Initiation of transcription of the mitochondrial genome of the trypanosomatid Crithidia oncopelti in vitro]. PMID- 2556256 TI - [Liquid crystalline dispersion of superhelical circular DNA as a base for biosensors]. PMID- 2556258 TI - [Thermostability of spatial organization of protein molecules and mechanisms of its support]. PMID- 2556259 TI - Mutations conferring resistance to quinol oxidation (Qz) inhibitors of the cyt bc1 complex of Rhodobacter capsulatus. AB - Several spontaneous mutants of the photosynthetic bacterium Rhodobacter capsulatus resistant to myxothiazol, stigmatellin and mucidin--inhibitors of the ubiquinol: cytochrome c oxidoreductase (cyt bc1 complex)--were isolated. They were grouped into eight different classes based on their genetic location, growth properties and inhibitor cross-resistance. The petABC (fbcFBC) cluster that encodes the structural genes for the Rieske FeS protein, cyt b and cyt c1 subunits of the cyt bc1 complex was cloned out of the representative isolates and the molecular basis of inhibitor-resistance was determined by DNA sequencing. These data indicated that while one group of mutations was located outside the petABC(fbcFBC) cluster, the remainder were single base pair changes in codons corresponding to phylogenetically conserved amino acid residues of cyt b. Of these substitutions, F144S conferred resistance to myxothiazol, T163A and V333A to stigmatellin, L106P and G152S to myxothiazol + mucidin and M140I and F144L to myxothiazol + stigmatellin. In addition, a mutation (aer126) which specifically impairs the quinol oxidase (Qz) activity of the cyt bc1 complex of a non photosynthetic mutant (R126) was identified to be a glycine to an aspartic acid replacement at position 158 of cyt b. Six of these mutations were found between amino acid residues 140 and 163, in a region linking the putative third and fourth transmembrane helices of cyt b. The non-random clustering of several inhibitor-resistance mutations around the non-functional aer126 mutation suggests that this region may be involved in the formation of the Qz inhibitor binding/quinol oxidation domain(s) of the cyt bc1 complex. Of the two remaining mutations, the V333A replacement conferred resistance to stigmatellin exclusively and was located in another region toward the C terminus of cyt b. The L106P substitution, on the other hand, was situated in the transmembrane helix II that carries two conserved histidine residues (positions 97 and 111 in R. capsulatus) considered to be the axial ligands for the heme groups of cyt b. The structural and functional roles of the amino acid residues involved in the acquisition of Qz inhibitor resistance are discussed in terms of the primary structure of cyt b and in relation to the natural inhibitor-resistance of various phylogenetically related cyt bc/bf complexes. PMID- 2556260 TI - Preferential, cooperative binding of DNA topoisomerase II to scaffold-associated regions. AB - DNA elements termed scaffold-associated regions (SARs) are AT-rich stretches of several hundred base pairs which are known to bind specifically to nuclear or metaphase scaffolds and are proposed to specify the base of chromatin loops. SARs contain sequences homologous to the DNA topoisomerase II cleavage consensus and this enzyme is known to be the major structural component of the mitotic chromosome scaffold. We find that purified topoisomerase II preferentially binds and aggregates SAR-containing DNA. This interaction is highly cooperative and, with increasing concentrations of topoisomerase II, the protein titrates quantitatively first SAR-containing DNA and then non-SAR DNA. About one topoisomerase II dimer is bound per 200 bp of DNA. SARs exhibit a Circe effect; they promote in cis topoisomerase II-mediated double-strand cleavage in SAR containing DNA fragments. The AT-rich SARs contain several oligo(dA).oligo(dT) tracts which determine their protein-binding specificity. Distamycin, which is known to interact highly selectively with runs of A.T base pairs, abolishes the specific interaction of SARs with topoisomerase II, and the homopolymer oligo(dA).oligo(dT) is, above a critical length of 240 bp, a highly specific artificial SAR. These results support the notion of an involvement of SARs and topoisomerase II in chromosome structure. PMID- 2556261 TI - Complex formation of human papillomavirus E7 proteins with the retinoblastoma tumor suppressor gene product. AB - The E7 proteins encoded by the human papillomaviruses (HPVs) associated with anogenital lesions share significant amino acid sequence homology. The E7 proteins of these different HPVs were assessed for their ability to form complexes with the retinoblastoma tumor suppressor gene product (p105-RB). Similar to the E7 protein of HPV-16, the E7 proteins of HPV-18, HBV-6b and HPV-11 were found to associate with p105-RB in vitro. The E7 proteins of HPV types associated with a high risk of malignant progression (HPV-16 and HPV-18) formed complexes with p105-RB with equal affinities. The E7 proteins encoded by HPV types 6b and 11, which are associated with clinical lesions with a lower risk for progression, bound to p105-RB with lower affinities. The E7 protein of the bovine papillomavirus type 1 (BPV-1), which does not share structural similarity in the amino terminal region with the HPV E7 proteins, was unable to form a detectable complex with p105-RB. The amino acid sequences of the HPV-16 E7 protein involved in complex formation with p105-RB in vitro have been mapped. Only a portion of the sequences that are conserved between the HPV E7 proteins and AdE1A were necessary for association with p105-RB. Furthermore, the HPV-16 E7-p105-RB complex was detected in an HPV-16-transformed human keratinocyte cell line. PMID- 2556262 TI - The role of ACE3 in Drosophila chorion gene amplification. AB - ACE3, an amplification control element for the third chromosome chorion cluster of Drosophila melanogaster, was identified previously as a cis-regulatory element for amplification of transposons containing the three chorion genes s18, s15 and s19. The deletion defining ACE3, located from -620 to -190 bp upstream of s18, disrupted both amplification and s18 transcription, suggesting that ACE3 might contain a transcription enhancer that regulated replication, as had been observed in a number of eukaryotic viruses. We show here that transcription control can be separated from replication control in delineating ACE3 to a 320 bp region. Addition of heterologous enhancers fails to activate amplification in tissues other than the follicle cells. Therefore ACE3 does not appear to be analogous to a transcription enhancer. However, further deletions within the ACE3 region revealed that it contains multiple functional domains. In addition, ACE3 functions independently of orientation with respect to other chorion sequences, and can be moved 1.5 kb away from other chorion sequences without eliminating amplification. PMID- 2556263 TI - 'SPKK' motifs prefer to bind to DNA at A/T-rich sites. AB - The termini of histone H1 and sea urchin spermatogenous H1 and H2B, which are essential for correct chromatin condensation, often contain repeats of the sequence SPK(R)K(R). A special type of beta-turn structural motif has been proposed for this sequence, and it has been shown that a segment of the sea urchin sperm H1 N terminus, which has six repeats of the motif (S6 peptide), binds to DNA and competes with the DNA binding drug Hoechst 33258. Here, we demonstrate by quantitative analysis of hydroxyl radical footprints that the synthetic oligopeptide, SPRKSPRK (S2), and the S6 peptide prefer to bind to the minor groove of DNA at the same A/T-rich sites. The locations of these binding sites are similar to Hoechst, but the sequence specificity of the oligopeptides is lower than that of Hoechst, and the detailed protection patterns differ slightly. We suggest that these small peptides and Hoechst recognize similar sequence-dependent features of the local architecture of DNA. PMID- 2556264 TI - The SV40 TC-II(kappa B) and the related H-2Kb enhansons exhibit different cell type specific and inducible proto-enhancer activities, but the SV40 core sequence and the AP-2 binding site have no enhanson properties. AB - The enhancer activity of the oligomerized SV40 TC-I and TC-II sequences has been investigated in lymphoid and non-lymphoid cell lines. While the TC-I sequence had no demonstrable enhanson activity, a class C enhanson (proto-enhancer), 5' GGAAAGTCCCC-3', overlapping the TC-II sequence and the GT-I enhanson was identified. This TC-II enhanson, which is identical to the kappa B motif from the kappa chain enhancer, was active in both lymphoid and non-lymphoid cells, which contrasts with the previously reported lymphoid cell specificity of the kappa B motif. However, its activity in non-lymphoid cells is in agreement with our previous reports describing the effect of mutations in the 'TC region' within the total SV40 enhancer in lymphoid and non-lymphoid cells. The activity of the TC-II enhanson could be moderately increased in HeLa by 12-O-tetradecanoyl-phorbol-13 acetate (TPA) and cycloheximide treatment, indicating that the protein(s) mediating its activity may be partially repressed by the previously described inhibitor protein I kappa B. The TC-II related, H-2Kb element, 5'-TGGGGATTCCCCA 3', of the histocompatibility class I H-2Kb gene promoter is also a class C enhanson which is active in both lymphoid and non-lymphoid cells. However, in contrast to the TC-II enhanson, the H-2Kb enhanson exhibits a very low activity in HeLa cells, but can be strongly induced by TPA and/or cycloheximide treatments which suggests that its cognate factor is inactivated (repressed) by an inhibitor protein. Interestingly, cycloheximide, but not TPA treatment, could induce the activity of both the TC-II and H-2Kb enhansons in F9 embryonal carcinoma cells, suggesting that these cells lack some component(s) of the protein kinase C signal transduction pathway. We also show that oligomers of the SV40 'core' sequence, which overlaps the TC-II enhanson, had no enhanson activity in any of the cell types studied, which questions the possible role of the AP-3 protein in SV40 enhancer activity in these cell types. In addition, oligomers of the AP-2 binding sites which are present in the SV40 TC region and in the human metallothionein IIA promoter show no enhanson activity, irrespective of whether the cells are treated with TPA.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2556266 TI - The octamer-binding proteins form multi-protein--DNA complexes with the HSV alpha TIF regulatory protein. AB - The herpes simplex virus transactivator, alpha TIF, stimulates transcription of the alpha/immediate early genes via a cis-acting site containing an octamer element and a conserved flanking sequence. The alpha TIF protein, produced in a baculovirus expression system, nucleates the formation of at least two DNA- protein complexes on this regulatory element. Both of these complexes contain the ubiquitous Oct-1 protein, whose POU domain alone is sufficient to allow assembly of the alpha TIF-dependent complexes. A second member of the POU domain family, the lymphoid specific Oct-2 protein, can also be assembled into similar complexes at high concentrations of alpha TIF protein. These complexes contain at least two cellular proteins in addition to Oct-1. One of these proteins is present in both insect and HeLa cells and probably recognizes sequences in the cis element. The second cellular protein, only present in HeLa cells, probably binds by protein protein interactions. PMID- 2556265 TI - The SV40 TC-II(kappa B) enhanson binds ubiquitous and cell type specifically inducible nuclear proteins from lymphoid and non-lymphoid cell lines. AB - We have characterized the complexes resulting from the specific binding in vitro of proteins present in nuclear extracts of several lymphoid and non-lymphoid cell lines to the TC-I and TC-II sequences of the simian virus 40 (SV40) enhancer. No proteins could be detected, binding selectively to the TC-I sequence, but two proteins TC-IIA and TC-IIB were identified interacting specifically with both the TC-II/kappa B enhanson, 5'-GGAAAGTCCCC-3' (important for the activity of the SV40 enhancer in vivo), and with the related H-2Kb enhanson, 5'-TGGGGATTCCCCA-3'. The binding of these two proteins to mutated TC-II enhansons correlates with the effect of these mutations in vivo, suggesting that both proteins may be important for SV40 enhancer activity. The TC-IIA binding activity was present in nuclear extracts of mature lymphoid B cells and was increased in pre-B cell nuclear extracts by lipopolysaccharide (LPS) and cycloheximide treatment. Furthermore, complex formation between the TC-IIA protein and the TC-II enhanson was efficiently competed by the kappa B motif from the kappa chain enhancer, indicating that TC-IIA is the NF-kappa B factor or a closely related protein. However, in contrast to previous reports, a TC-IIA/NF-kappa B-like protein whose properties could not be distinguished from those of the TC-IIA protein present in lymphoid B cells, was found in nuclear extracts of several untreated non-lymphoid cell lines, notably of HeLa cells, but not of undifferentiated F9 embryonal carcinoma (EC) cells [F9(ND)]. The TC-IIA binding activity which was moderately increased in HeLa cell nuclear extracts by 12-O-tetradecanoylphorbol-13-acetate (TPA) and/or cycloheximide treatment could be induced in nuclear extracts of F9(ND) cells by cycloheximide, but not by TPA. Moreover, the TC-IIA binding activity could be induced in cytosolic fractions from F9(ND) cells by treatment with deoxycholate, indicating that these cells contain an inhibitor protein similar to the previously described NF-kappa B inhibitor, I kappa B. The second TC-II enhanson binding protein, TC-IIB, which could be clearly distinguished from the TC-IIA/NF-kappa B-like protein, by a number of differential properties, resembles the previously described KBF1/H2TF1 protein as it binds with a higher affinity to the H-2Kb enhanson than to the TC-II/kappa B enhanson, and its pattern of methylation interference on the H-2Kb and TC-II/kappa B enhansons is identical to that reported for the KBF1/H2TF1 protein.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2556267 TI - NF-kappa B activation of the cytomegalovirus enhancer is mediated by a viral transactivator and by T cell stimulation. AB - The expression of cytomegalovirus alpha (immediate early) genes is under control of an enhancer that carries signals for strong constitutive expression as well as response elements for transactivation by viral proteins. We have used synthetic oligonucleotides representing the 16, 18 and 19 bp repeat elements within the enhancer to investigate the role of virus-induced cellular transcription factors in enhancer activation. We show that the transcription factor NF-kappa B, which binds to the 18 bp repeat, plays a central role in enhancer activation in infected human fibroblasts and that activation is mediated by the product of the viral gene ie1. The simian immunodeficiency virus kappa B site can functionally substitute for the 18 bp element in transient transactivation assays and can also compete efficiently for specific binding to the 18 bp repeat element in vitro. Point mutations in the NF-kappa B site within the 18 bp element disrupt ie1 mediated transactivation and binding. We have found that the characteristics of the 18 bp binding factor from human fibroblasts are indistinguishable from NF kappa B induced by phorbol ester plus mitogen treatment of T lymphocytes, as determined by gel mobility shift assay as well as protection of the binding site from chemical cleavage. Furthermore, T cell stimulation mediates activation of the viral enhancer via kappa B sites, an observation that may be important in the interaction of cytomegalovirus with the naturally infected human host. Thus, NF kappa B plays a central role as a target for enhancer activation via viral and cellular factors. PMID- 2556268 TI - Cruciform cutting endonucleases from Saccharomyces cerevisiae and phage T4 show conserved reactions with branched DNAs. AB - We have purified a cruciform DNA resolving endonuclease (Endo X3) greater than 1000-fold from crude extracts of mitotically growing Saccharomyces cerevisiae. The enzyme shows high specificity for DNAs with secondary structures and introduces characteristic patterns of staggered 'nicks' in the immediate vicinity of the structure. The following substrates were analyzed in detail: (i) naturally occurring four-way X junctions in cruciform DNA of a supercoiled plasmid; (ii) synthetic four-way X junctions with arms of 9 bp; (iii) synthetic three-way Y junctions with arms of 10 bp; and (iv) heteroduplex loops with 19 nucleotides in the loop. Cleavages were always found in the double stranded portion of the DNA, located immediately adjacent to the junction of the respective structure. The Endo X3 induced cleavage patterns are identical or very similar to the cleavage patterns induced in the same substrates by endonuclease VII (Endo VII) from phage T4. Furthermore, the activity of Endo X3 is completely inhibited in the presence of anti-Endo VII antiserum. Endo X3 has an apparent mol. wt of 43,000 daltons, determined by gel filtration and of approximately 18,000 daltons in SDS- polyacrylamide gels. Maximum activity of the enzyme was obtained in the presence of 10 mM MgCl2 at 31 degrees C in Tris-HCl buffer over a broad pH range with a maximum approximately 8.0. About 70% of maximal activity was obtained when Mg2+ was replaced by equimolar amounts of Mn2+ or Ca2+. PMID- 2556270 TI - Hydrodynamic, structural and magnetic properties of Megasphaera elsdenii Fe hydrogenase reinvestigated. AB - Megasphaera elsdenii hydrogenase has been purified to homogeneity using an FPLC procedure as the final step. The protein gives a single band in SDS/PAGE with an apparent molecular mass of 57-59 kDa. There is no second hydrogenase activity in the soluble fraction of M. elsdenii. The hydrodynamics of the enzyme have been compared to those of the two-subunit Fe hydrogenase from Desulfovibrio vulgaris (Hildenborough) in the analytical ultracentrifuge using the absorption of the intrinsic iron-sulfur clusters as the monitor. Sedimentation-velocity experiments indicate the M. elsdenii enzyme (s20,w = 4.95 S) to be essentially globular, while the D. vulgaris enzyme (s20,w = 4.1 S) has a less symmetric shape. From the sedimentation equilibrium measurements under a variety of conditions an average molecular mass is calculated of 58 kDa (M. elsdenii) and 54 kDa (D. vulgaris), respectively. Pure, maximally active M. elsdenii hydrogenase has A405/A280 = 0.36 and has a specific H2-production activity of 400 mumol H2.min-1.(mg protein)-1 at 30 degrees C and pH 8.0. The enzyme contains some 13-18 iron and acid-labile sulfur ions/58-kDa monomer. Eight of these Fe-S are present as two electron transferring ferredoxin-like cubanes with Em approximately greater than -0.3 V, as indicated by pH-dependent EPR spectroscopy on the H2-reduced enzyme. In the (re)oxidized state the remainder iron gives rise to a single S = 1/2 rhombic EPR signal. Hydrogen-production activity, content of remainder iron and rhombic EPR signal intensity are mutually correlated. Purified hydrogenase appears to exist as a mixture of fully active holoenzyme and inactive protein still carrying the two cubanes but deficient in active-site iron. PMID- 2556271 TI - NADPH-dependent H2O2 generation catalyzed by thyroid plasma membranes. Studies with electron scavengers. AB - Hog thyroid plasma membrane preparations containing a Ca2+-regulated NADPH dependent H2O2-generating system were studied. The Ca2+-dependent reductase activities of ferricytochrome c, 2,6-dichloroindophenol, nitroblue tetrazolium, and potassium ferricyanide were tested and the effect of these scavengers on H2O2 formation, NADPH oxidation and O2 consumption were measured, with the following results. 1. Thyroid plasma membrane Ca2+-independent cytochrome c reduction was not catalyzed by the NADPH-dependent H2O2-generating system. This activity was superoxide-dismutase-insensitive. 2. Of the three other electron scavengers tested, only K3Fe(CN)6 was clearly, but partially reduced in a Ca2+-dependent manner. 3. Though the NADPH-dependent reduction of nitroblue tetrazolium was very low and superoxide-dismutase-insensitive, nitroblue tetrazolium inhibited O2 consumption, H2O2 formation and NADPH oxidation, indicating that nitroblue tetrazolium inhibits the H2O2-generating system. We conclude that the thyroid plasma membrane H2O2-generating system does not or liberate O2- and that Ca2+ controls the first step (NADPH oxidation) of the H2O2-generating system. PMID- 2556269 TI - The DNA unwinding element: a novel, cis-acting component that facilitates opening of the Escherichia coli replication origin. AB - We have discovered that DNA supercoiling, in the absence of replication proteins, induces localized unwinding in the Escherichia coli replication origin (oriC) at the same sequence opened by the dnaA initiator protein. The DNA helix at the tandemly repeated, 13mer sequence is thermodynamically unstable, as evidenced by hypersensitivity to single-strand-specific nuclease in a negatively supercoiled plasmid, and demonstrated by stable DNA unwinding seen after two-dimensional gel electrophoresis of topoisomers. A replication-defective oriC mutant lacking the leftmost 13mer shows no nuclease hypersensitivity in two remaining 13mers and no detectable DNA unwinding on two-dimensional gels. The replication defect in the oriC mutant can be corrected by inserting a dissimilar DNA sequence with reduced helical stability in place of the leftmost 13mer. Thus, the helical instability of the leftmost 13mer, not the specific 13mer sequence, is essential for origin function. The rightmost 13mer exhibits helical instability but differs from the leftmost 13mer in its strict sequence conservation among related bacterial origins. The repeated 13mer region appears to serve two overlapping functions: protein recognition and helical instability. We propose that the cis-acting sequence whose helical instability is required for origin function be called the DNA unwinding element (DUE). PMID- 2556272 TI - A distinct form of ribonuclease H from calf thymus stimulates its homologous DNA polymerase-alpha-primase complex. AB - A ribonuclease H which degrades RNA specifically in RNA-DNA hybrids and, moreover, stimulates its homologous DNA-polymerase-primase complex was purified from calf thymus. The enzyme consists of a single polypeptide of molecular mass 78 kDa. It requires divalent cations for activity, and prefers Mg2+ over Mn2+. Ribonuclease H is optimally active at neutral pH and in 75 mM potassium acetate and is strongly sensitive to N-ethylmaleimide. [3H]Poly(rA).poly(dT), [3H]poly(rC).poly(dI), and [3H]RNA.M13-DNA are degraded to 3-9-mer oligoribonucleotides with similar kinetics, whereas double- or single-stranded DNA, and double- and single-stranded RNA remain unaffected. The enzyme stimulates in vitro DNA synthesis by the immunoaffinity-purified calf-thymus DNA-polymerase alpha-primase complex threefold. When ribonuclease H is present in a three-fold molar excess to the polymerase-primase complex, twice as much primer is formed as in the absence of ribonuclease H. Ribonuclease H also stimulates the elongation rate of DNA polymerase alpha by a factor of 2-3, independent of whether primase primed DNA templates or templates primed with oligonucleotides are used. Our results suggest that this form of ribonuclease H is a likely candidate for a genuine primer-removing enzyme in mammalian cells. PMID- 2556273 TI - Electron transport pathways to nitrous oxide in Rhodobacter species. AB - 1. Electron transport components involved in nitrous oxide reduction in several strains of Rhodobacter capsulatus and in the denitrifying strain of Rhodobacter sphaeroides (f. sp. denitrificans) have been investigated. Detailed titrations with antimycin A and myxothiazol, inhibitors of the cytochrome bc1 complex, show that part of the electron flow to nitrous oxide passes through this complex. The sensitivity to myxothiazol varies between strains and growth conditions of R. capsulatus; the higher rates of nitrous oxide reduction correlate with the higher sensitivities. Partial inhibition of the nitrous oxide reductase enzyme with azide decreased the sensitivity to myxothiazol of the strains that had the highest nitrous oxide reductase activity. 2. Inhibition of nitrous oxide reduction in cells of R. capsulatus by myxothiazol could be restored under dark conditions by addition of N,N,N',N'-tetramethyl-p-phenylene diamine. The highest activities observed after addition of this electron carrier were found in the strains that had the highest sensitivity to myxothiazol, consistent with the premise that this inhibitor is more effective at the higher flux rates to nitrous oxide. 3. Addition of nitrous oxide to cells of R. capsulatus strain N22DNAR+ under darkness caused oxidation of both b- and c-type cytochromes. The oxidation of b cytochromes was less pronounced in the presence of myxothiazol, consistent with a role for the cytochrome bc1 complex in the electron pathway to nitrous oxide. Ferricyanide, in the absence of myxothiazol, caused a similar extent of oxidation of b cytochromes, but a greater oxidation of c-type, suggesting that there was a pool of c-type cytochrome that was not oxidisable by nitrous oxide. The time course showed that both the b- and c-type cytochromes were oxidised within a few seconds of the addition of nitrous oxide. During the following seconds there was a partial re-reduction of the cytochromes such that after approximately 1 min a lower steady-state of oxidation was attained and this persisted until the nitrous oxide was exhausted. 4. A mutant, MTCBC1, of R. capsulatus that specifically lacked a functional cytochrome bc1 complex reduced nitrous oxide, albeit at 30% of the rate shown by the parent strain MT1131. A reduced minus nitrous-oxide-oxidised difference spectrum for MTCBC1 in the absence of myxothiazol was similar to the corresponding difference spectrum observed for strain N22DNAR+ in the presence of myxothiazol. It is suggested that these difference spectra identify the cytochrome components, including a b-type, involved in a pathway that is alternative to, and independent of, the cytochrome bc1 complex.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2556274 TI - Neomycin induces high-affinity agonist binding of G-protein-coupled receptors. AB - Neomycin, an inositol-phospholipid-binding aminoglycoside antibiotic, is known to interfere with signal transduction mechanisms involving phospholipase C as effector enzyme. In this study, we report that neomycin can also markedly influence agonist binding of G-protein-coupled receptors. In membranes of differentiated human leukemia cells (HL 60 cells), neomycin (0.1-10 mM) was found to induce high-affinity binding of the chemotactic tripeptide, N-formyl methionylleucylphenylalanine (fMet-Leu-Phe), to its receptor sites in a manner similar to magnesium. Gentamycin and streptomycin, two other aminoglycoside antibiotics, were as potent and as effective as neomycin or magnesium in inducing high-affinity agonist receptor binding. Pretreatment of the cells with pertussis toxin reduced the effects of magnesium and neomycin on agonist receptor binding likewise. In contrast, magnesium but not neomycin largely enhanced the potency of guanine nucleotides, particularly of GTP and its analog, guanosine-5'-O-(3 thiotriphosphate), to reduce fMet-Leu-Phe receptor binding, while maximal inhibition of agonist receptor binding by guanine nucleotides was identical with magnesium and neomycin. Furthermore, neomycin could not replace magnesium in providing stimulation of HL 60 membrane high-affinity GTPase by fMet-Leu-Phe. In close agreement to these findings on the pertussis-toxin-sensitive Gi-protein coupled formyl peptide receptors, neomycin in a manner similar to magnesium induced high-affinity agonist binding of Gs-protein-coupled beta-adrenoceptors. Similar to formyl peptide receptor binding, high-affinity binding of isoproterenol to beta-adrenoceptors in guinea pig lung membranes induced by magnesium and neomycin was inhibited by the GTP analog, guanosine-5'-O-(3 thiotriphosphate), to a similar maximal extent but with an about 100-fold higher potency in the presence of magnesium than in the presence of neomycin. The data presented thus indicate that neomycin and other aminoglycoside antibiotics can mimic the action of magnesium (or other divalent cations) in inducing high affinity agonist binding of Gi- and Gs-protein-coupled receptors, but not in inducing subsequent G-protein activation by guanosine triphosphates. The data, furthermore, suggest that neomycin by this selective action will be a powerful tool to dissect the multiple sites of magnesium's action in the agonist receptor G-protein interaction. PMID- 2556276 TI - Fluconazole therapy in an underweight infant. PMID- 2556275 TI - Electron transport in sulfate-reducing bacteria. Molecular modeling and NMR studies of the rubredoxin--tetraheme-cytochrome-c3 complex. AB - A hypothetical model of the complex formed between the iron-sulfur protein rubredoxin and the tetraheme cytochrome c3 from the sulfate-reducing bacteria Desulfovibrio vulgaris (Hildenborough) has been proposed utilizing computer graphic modeling, computational methods and NMR spectroscopy. The proposed complex appears feasible on the basis of complementary electrostatic interaction and steric factors and is consistent with the data from NMR experiments. In this model, the non-heme iron atom of rubredoxin is in close proximity to heme 1 of cytochrome c3. The complex is stabilized by charge-pair interactions and hydrogen bonds. This complex is compared to the flavodoxin-cytochrome c3 complex previously proposed [Stewart, D. E., LeGall, J., Moura, I., Moura, J. J. G., Peck, H. D. Jr, Xavier, A. V., Weiner, P. K. & Wampler, J. E. (1988) Biochemistry 27, 2444-2450] and new NMR data shows that both proteins interact with the same heme group of the cytochrome as postulated. PMID- 2556278 TI - Ascites related to cytomegalovirus infection. PMID- 2556277 TI - In vitro activity of combinations of beta-lactam antibiotics with beta-lactamase inhibitors against cephalosporinase-producing bacteria. AB - Combinations of different beta-lactam antibiotics, including cefotaxime, with three beta-lactamase inhibitors were tested against cephalosporinase producing bacterial strains. The most significant antagonism was obtained with a combination of clavulanic acid and cefotaxime, while almost no antagonism was observed with sulbactam and tazobactam. In strains belonging to five different species there was a correlation between the levels of cephalosporinase produced after exposure to different concentrations of inhibitors and the MICs of cefotaxime combined with the same concentrations of inhibitors. It is concluded that there is little likelihood of antagonism between beta-lactam antibiotics and sulbactam or tazobactam. PMID- 2556279 TI - Absorption of oral enalapril in germ-free and microbially-associated rats. AB - The effect of microbial status and presence of food in the gastrointestinal (GI) tract on absorption of an orally administered antihypertensive drug [14C] enalapril (1 mg/10 microCi/kg body weight) was studied in non-fasted and fasted germ-free (GF) and microbially-associated (MA) rats. The absorption was evaluated from blood samples taken 0.5, 2 and 4 h after the administration of enalapril. Drug levels in the lungs, liver, and duodenal mucosa were also measured by counting [14C]-radioactivities at 4 h after the drug administration. During the 4 h monitoring period enalapril was poorly absorbed in non-fasted rats as compared to fasted animals. In non-fasted rats, the absorption of enalapril was slightly slower in GF than in MA rats. In fasted rats, no difference was found between GF and MA animals. The results show that food in the GI tract substantially lowers or retards the absorption of enalapril in rats. The GF status of the GI tract, however, did not substantially affect the absorption of the ester-type drug enalapril. PMID- 2556280 TI - Relation of in vitro drug activity to clinical response in a prospective trial for advanced germ cell testicular tumors. AB - An in vitro assay, which evaluates the effect of drugs on labelled DNA precursor incorporation into fresh specimens after 3 h of in vitro treatment, was used for a prospective study. Drugs were used as monochemotherapy or in different combinations according to their in vitro activities for salvage or palliation treatment of 16 patients with germ cell testicular tumors, for a total of 21 correlations between in vitro and clinical results. In 11 instances, drugs active in vitro were used; in 10 instances, because no drug was active in vitro, patients were treated with inactive drugs. There was a significant correlation between in vitro and clinical activity (p = 0.02), with a very high true-negative rate (90%) and a lower true-positive rate (54%). No patient treated with drugs inactive in vitro survived 2 years, whereas freedom from progression and overall survival for patients treated with drugs active in vitro were 27.3 and 36.4%, respectively (active vs. inactive, p = 0.007). The overall response in the group of patients treated prospectively was comparable to that observed in a group of 9 patients treated in the same period of time with drugs of unknown in vitro activity (33 vs. 11% objective and long-term responses, respectively, p = n.s.) notwithstanding the high percentage of patients treated with drugs inactive in vitro in the former group. PMID- 2556281 TI - Phase II study of pirarubicin in advanced non-small cell lung cancer. PMID- 2556282 TI - Conservative treatment of early breast cancer: prognostic value of the ductal in situ component and other pathological variables on local control and survival. Long-term results. AB - Four hundred and thirty-four patients with infiltrative ductal carcinoma were treated by limited surgery and irradiation between January 1960 and December 1980. The median follow-up was 103 months. Retrospective pathological analysis of the primary tumor identified a subset of pathological parameters which were predictors of local breast failure and survival. Pathological predictors of local breast recurrence were: incomplete surgical excision (P less than 0.0001), lymphatic invasion (P less than 0.02) and presence of an extensive in situ component (EDISC) (P less than 0.03). Pathological predictors of survival were: incomplete surgery (P less than 0.007), size of the primary tumour (P less than 0.03), high histologic grade (P less than 0.005), lymphatic invasion (P less than 0.0001) and absence of associated in situ component (P less than 0.008). This study emphasizes the role of the in situ component in the prognosis of breast carcinoma treated with conservative management. PMID- 2556283 TI - Beneficial hemodynamic effects of milrinone and enalapril in conscious rats with healed myocardial infarction. AB - Milrinone and enalapril, which inhibit PDE-III and ACE, respectively, are able to prolong survival of myocardially infarcted (MI) rats. This study sought to identify oral hemodynamic effects of these agents which could underlie such efficacy in this heart failure model. Four weeks after ligation of the left main coronary artery, basal left ventricular (LV) systolic pressure and dP/dtmax, heart rate and mean blood pressure of the MI rats were significantly less than that of sham-operated controls, and LV end-diastolic pressure (LVEDP) was markedly elevated. Milrinone, at 2.0 mg/kg, reduced LVEDP and renal blood flow of these 4-week MI rats by an average of 39 and 18%, respectively (P less than 0.05) within 1 h. At 4.0 mg/kg, it reduced LVEDP by 46% and raised heart rate by 16% (P less than 0.05). Enalapril (1.0 mg/kg) increased small intestine blood flow of these compromised rats by 16% (P less than 0.05), and tended to reduce LVEDP ( 28%) within 1.5 h. Treatment with milrinone (2.0 mg/kg) plus enalapril (1.0 mg/kg) promoted LV dP/dtmax, coronary blood flow, and heart rate by 48, 40 and 13%, and reduced LVEDP by 40% (P less than 0.05 for all effects). Thus these agents can reduce LVEDP and redistribute cardiac output of MI rats. Furthermore, the combination of enalapril and milrinone can restore LVEDP and LV dP/dtmax of MI rats to near normal and promote coronary blood flow without compromising cardiac output or renal blood flow. Such effects, it timely or sustained, may prolong survival. PMID- 2556284 TI - Mu opiate receptors are selectively labelled by [3H]carfentanil in human and rat brain. AB - [11C]Carfentanil is a potent opioid agonist currently in use as a specific PET (position emission tomography) scan radioligand for brain mu opioid receptors. In order to investigate the receptor interactions of carfentanil in detail [3H]carfentanil was used as a radioligand for labelling receptors in rat and human brain tissue homogenates. [3H]Carfentanil was found to bind saturably and with high affinity (KD = 0.08 +/- 0.01 nM) to membranes prepared from human cortical (Bmax = 42 +/- 3 fmol/mg) and thalamic (Bmax = 84 +/- 3 fmol/mg) tissues and rat cortex (Bmax = 82 +/- 4 fmol/mg) and diencephalon (Bmax = 105 +/- 5 fmol/mg). Association (1.23 +/- 0.19 X 10(10) Mol-1 X min-1 and dissociation rate (0.19 +/- 0.03 min-1) constants were determined in human cortical tissues; results from studies in rat cortical, and rat diencephalon tissue homogenates produced similar kinetic rate constants. Competition studies with a variety of drugs indicated that [3H]carfentanil interacts primarily with mu opioid receptors in the four tissues studied; the affinities of a series of non-radioactive opioid ligands were essentially identical in the four tissues (correlation coefficients = 0.88-0.93). Naloxone, morphine, DAGO [( D-Ala2-MePhe4-Gly-ol5]enkephalin), DADL [( D-Ala2-D-Leu5]enkephalin) and EKC (ehtylketazocine) potently displaced specific [3H]carfentanil binding with nM potency while the kappa agonist U-69593, the sigma agonists (+)-SKF 10047, (+)-3-PPP [3-hydroxyphenyl)-N-propylpiperidine) and haloperidol and PCP (phencyclidine) were less potent displacing agents. The higher affinities of DAGO and morphine versus DADL for the [3H]carfentanil binding sites indicates that delta opioid receptors are not being labelled.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556285 TI - Desmethoxyverapamil-sensitive calcium channels in rat portal vein smooth muscle. AB - The whole-cell patch clamp technique was used to analyze the properties of the phenylalkylamine-sensitive calcium channels in smooth muscle cells isolated from the portal vein. (-)-D888 dose dependently inhibited the calcium current elicited from a holding potential of -40 mV (IC50 = 1.3 nM) in a frequency-dependent manner. No voltage dependence of the inhibition was noted. Independent high- and low-affinity binding sites for (-)-[3H]D888 were identified. Calcium entry blockers such as (-)-D888, d-cis-diltiazem and nicardipine completely or partially antagonized the (-)-[3H]D888 binding at both types of sites. The properties of this cross-inhibition suggest that phenylalkylamines and d-cis diltiazem bind at common sites in vascular smooth muscles whereas dihydropyridines bind at distinct sites which are allosterically coupled to the phenylalkylamine sites. As the IC50 for (-)-D888 found from electrophysiological experiments is not identical to the equilibrium dissociation constants for the high- and low-affinity sites found from binding data (0.47 and 50 nM, respectively), it is suggested that binding of (-)-D888 to both high- and low affinity sites may be involved in the inhibitory effect of (-)-D888 on calcium channels. Furthermore, these two different binding sites may correspond to two different subtypes of phenylalkylamine-sensitive calcium channels in smooth muscle cells. PMID- 2556286 TI - Specific high-affinity binding sites for 125I-labelled porcine endothelin in rat cardiac membranes. AB - 125I-labelled porcine endothelin (125I-endothelin) was used to identify specific high affinity endothelin binding sites in rat cardiac membrane fragments. Binding was to a single population of sites, with a KD of 0.20 +/- 0.03 nM and a Bmax of 93.5 +/- 6.4 fmol/mg protein at 37 degrees C. Reducing the temperature to 25 degrees C increased (P less than 0.02) the KD without changing Bmax. 125I Endothelin binding was Ca2+ independent. Specific binding was saturable and displaceable by cold endothelin and sarafotoxin S6b, but not by (-)Bay K8644, nicardipine, (-)D888, (+)cis-diltiazem, prenylamine, lidoflazine, flunarizine, nor by 10(-10)-10(-4) M CoCl2, nor 10(-10)-10(-4) M NiCl2. omega-Conotoxin, prazosin, isoprenaline, angiotensin II and its inhibitor, vasopressin and its inhibitor, glyceryl trinitrate, amiloride, ergometrine and FII stonefish toxin also failed to displace bound 125I-endothelin. 10(-4)-10(-2) M CaCl2, 10(-4)-10( 2) M MgCl2, 3 X 10(-6)-10(-3) M MnCl2, 10(-5)-3 X 10(-4) M NiCl2, and 3 X 10(-5) 3 X 10(-4) M CoCl2 stimulated the binding. Incubation at 100 degrees C for 10 min destroyed specific binding. PMID- 2556287 TI - Glycine antagonist action of 1-aminocyclobutane-1-carboxylate (ACBC) in Xenopus oocytes injected with rat brain mRNA. AB - ACBC has been reported to have the binding profile of an antagonist at the glycine site of the NMDA receptor. In Xenopus oocytes injected with rat brain mRNA, we have confirmed the antagonist action of ACBC on NMDA responses. ACBC and HA-966, a known glycine antagonist, blocked NMDA responses in a non-competitive manner and blocked the potentiation of NMDA responses by glycine in a competitive manner. We conclude that ACBC blocks NMDA responses via a competitive interaction at the glycine modulatory site. PMID- 2556288 TI - Methoctramine, a cardioselective muscarinic antagonist, stimulates phosphoinositide hydrolysis in rat cerebral cortex. AB - The cardioselective muscarinic antagonist methoctramine antagonized carbamylcholine-mediated phosphoinositide (PI) hydrolysis in a concentration dependent fashion in dissociated rat cerebrocortical cells. However, as the concentration of methoctramine was increased above 5 microM, there was a reversal of the antagonism of the PI response. In the absence of carbamylcholine, methoctramine by itself significantly increased PI hydrolysis with a maximal effect at 30 microM. Various classes of receptor antagonists, including atropine, and ion-channel blockers were unable to block methoctramine-stimulated PI hydrolysis. PMID- 2556289 TI - Studies on the role of the NMDA receptor in the substantia nigra pars reticulata and entopeduncular nucleus in the development of the high pressure neurological syndrome in rats. AB - The effect of the focal injection of N-methyl-D-aspartate (NMDA) and 2-amino-7 phosphonoheptanoate (APH) into the substantia nigra pars reticulata (SNR) and entopeduncular nucleus (EP) on behavioural signs of the high pressure neurological syndrome (HPNS) in rats was studied. Doses of 1, 5 and 10 nmoles of NMDA or APH were injected into the SNR or EP, 10-30 min prior to the exposure of animals to a high pressure. Injection of NMDA into either SNR or EP results in a lowering of the threshold pressure for tremor by about 30%. Injection of NMDA into the SNR has no significant effect on clonic seizures whereas its injection into the EP results in a decrease of threshold pressure for clonic seizures. NMDA also facilitates the occurrence of forelimb clonus when injected into the EP. Injection of the NMDA antagonist, APH, into the SNR or EP significantly increases the threshold pressure of tremor (32.8 and 48.2% respectively). Seizure threshold is also increased by the injection of APH into either area, but nigral injections (especially the higher doses) are more protective against seizures than the EP injections. Comparing the two sites blockade of NMDA receptors within the EP is more protective against tremor, whereas in the SNR NMDA blockade is more protective against seizures. PMID- 2556291 TI - Localization of nerve growth factor receptor messenger RNA and protein in the adult rat brain. AB - We have used in situ hybridization and immunocytochemistry to map the cellular localization of NGF receptor (NGF-R) mRNA and protein in the adult rat brain. In addition to basal forebrain magnocellular neurons, NGF-R is widely expressed within the CNS, including neurons of the caudate/putamen, ventral premamillary nucleus, mesencephalic trigeminal nucleus, prepositus hypoglossal nucleus, raphe nucleus, nucleus ambiguous, and Purkinje cells of the cerebellum. Cells of the vestibulocochlear ganglion also contain NGF-R mRNA and protein. Ventricular subependymal cells and tanycytes are clearly stained by immunocytochemistry, yet only very weak hybridization is detectable in these cells. Also, greater amounts of NGF-R protein than of mRNA appear to be present in the glomeruli of the olfactory bulb, area postrema, and nucleus tractus solitarius. Areas that contain only NGF-R immunoreactive fibers and terminals can be distinguished from the cellular sites of NGF-R biosynthesis and include the suprachiasmatic nucleus, the principal olivary pretectal nucleus, the superior colliculus, the inferior olive, and the principal and spinal trigeminal nuclei. This study shows that NGF-R is widely expressed within individual neurons in different areas of the rat brain and identifies new potential CNS target sites of endogenous NGF. PMID- 2556290 TI - Excitatory amino acid receptors in the caudal ventrolateral medulla mediate a vagal cardiopulmonary reflex in the rat. AB - The importance of the caudal ventrolateral medulla (CVLM) in mediating vagal cardiopulmonary (Bezold-Jarisch reflex) reflex activity was studied in urethane anaesthetized rats. Unilateral electrolytic lesion of the CVLM markedly attenuated Bezold-Jarisch reflex responses (hypotension and bradycardia) elicited by intravenous injections of 5-HT. Bilateral lesion of the CVLM virtually abolished the reflex responses. Microinjection of the excitatory amino acid (EAA) receptor antagonist kynurenate (KYN), but not the inactive analogue xanthurenate, into the CVLM markedly attenuated the reflex responses to 5-HT. The N-methyl-D aspartate (NMDA) receptor antagonist, MK-801 also markedly attenuated reflex activity. Furthermore, lesions, KYN and MK-801 all tended to elevate resting blood pressure and to reduce resting heart rate. These findings support the hypothesis that the CVLM is an important medullary locus mediating cardiovascular reflex integration and that an EAA synapse in the CVLM is important in the cardiopulmonary reflex arc. PMID- 2556292 TI - NGF receptor gene expression is decreased in the nucleus basalis in Alzheimer's disease. AB - Basal forebrain neuronal atrophy in Alzheimer's disease (AD) may be caused by a deficit in the NGF responsiveness of magnocellular cholinergic neurons which project to the cerebral cortex and hippocampal formation. We have used in situ hybridization to show that NGF-receptor (NGF-R) mRNA-positive neurons are lost within all divisions of the nucleus basalis of Meynert (Ch4 cell group) in AD patients as compared to normal aged controls. The posterior division of the nucleus basalis showed the largest decrease in NGF-R mRNA hybridization in the disease, with no overlap in neuronal number between AD cases and normal controls. Northern (RNA) blotting showed decreased levels of NGF-R mRNA in the nucleus basalis in the disease. No differences in the number of NGF-R mRNA-positive neurons between normal aged and AD patients were detected within the NGF responsive cell groups of the medial septum (Ch1) and nucleus of the vertical limb of the diagonal band (Ch2). These results show that NGF-R gene expression is selectively reduced within basal forebrain neuronal populations which exhibit degenerative changes in AD. PMID- 2556294 TI - Identification of a small intracellular region of the muscarinic m3 receptor as a determinant of selective coupling to PI turnover. AB - Molecular cloning studies have demonstrated the existence of five different muscarinic receptors (m1-m5). While m1, m3 and m5 strongly couple to stimulation of phosphoinositide (PI) hydrolysis, m2 and m4 are more efficiently linked to inhibition of adenylate cyclase. The sequences of m1-m5 have a short segment at the N-terminal portion of the putative third cytoplasmic loop (i3) which is highly conserved among m1, m3 and m5, but different from the sequence which is well conserved among m2 and m4. To study the role of this region in conferring coupling selectivity, we constructed cDNAs encoding chimeric m2/m3 receptors. Transient expression of these receptor hybrids in COS-7 cells showed that a 17 amino acid segment at the N-terminal portion of i3 is a major determinant of how efficiently the different muscarinic receptors are coupled to PI hydrolysis. PMID- 2556293 TI - Sequence and expression of a novel GABAA receptor alpha subunit. AB - Cloned cDNA encoding the bovine alpha 4 subunit of the GABAA receptor has been isolated. The predicted 51 amino acid long mature protein contains an exceptionally long intracellular domain and shares 53-56% sequence similarity to the previously characterized alpha 1, alpha 2 and alpha 3 subunits. Co-expression of alpha 4 and beta 1 in Xenopus oocytes resulted in the formation of GABA-gated chloride channels with expected pharmacology, although no benzodiazepine potentiation was observed. Northern analysis indicates that a 4 kb alpha 4 mRNA is expressed in the calf cerebellum, cortex and hippocampus but is barely detectable in the rat brain. PMID- 2556295 TI - Expression of the E. coli Lac Z gene from a defective HSV-1 vector in various human normal, cancer-prone and tumor cells. AB - Introducing foreign genetic material into human cells is essential for the elucidation of the function of various human genes and has potential use in the treatment of human diseases by gene therapy. In this study we demonstrate that a defective herpes simplex virus type 1 vector, pHSVlac, can effectively transfer and express the Escherichia coli Lac Z gene in a variety of exponential and quiescent human cells. The human cells tested included representative cells derived from cancer-prone patients that presumably have various DNA repair deficiencies. PMID- 2556297 TI - Interaction of the LexA repressor and the uvrC regulatory region. AB - We have studied the in vitro interaction of the LexA repressor protein and the uvrC regulatory region. We find that there is specific binding to two regions, the region we have defined as lexA1 and the lexA2-lexA3 region. Our findings support the possibility of an inducible regulation for this complex operon. PMID- 2556296 TI - Pharmacological characterization and region-specific expression in brain of the beta 2- and beta 3-subunits of the rat GABAA receptor. AB - The cDNA for a third beta-subunit of the rat GABAA receptor has been cloned using another beta-subunit, which we had previously cloned [(1989) FEBS Lett. 246, 145 148], as a probe. The approximately 8-kb cDNA for this beta-subunit (termed beta 2) encodes a protein of 474 amino acid residues that shares approximately 80% sequence identity with the rat and bovine beta 1- and beta 3-subunits. Coexpression of the cloned beta-subunit cDNA with the alpha 1-subunit cDNA of the rat GABAA receptor in Xenopus oocytes produced a functional receptor and Cl- channel with pharmacological characteristics of a GABAA receptor. In contrast to interchanging alpha-subunits [(1988) Nature 335, 76-79], exchange of beta 2- or beta 3-subunits in an alpha 1/beta receptor complex did not markedly alter the pharmacological properties of expressed receptors. In situ hybridization histochemistry with synthetic subunit-specific oligo-deoxynucleotide probes revealed a region-specific expression of alpha 1-, beta 2- and beta 3-subunit mRNAs in the rat central nervous system. These observations provide an additional molecular basis for the functional heterogeneity in the GABAA receptor complex. PMID- 2556299 TI - A consensus sequence for substrate hydrolysis by rhinovirus 3C proteinase. AB - Kinetic constants were determined for the hydrolysis of a series of synthetic peptide substrates by recombinant rhinovirus (HRV 14) 3C proteinase. Systematic removal or replacement of individual residues indicated that the minimum sequence required for effective cleavage by the viral cysteine proteinase was P5-Val/Thr P3-P2-Gln-Gly-Pro. PMID- 2556298 TI - Identification and signal transduction mechanism of elastin peptide receptor in human leukocytes. AB - The existence of a novel receptor on human polymorphonuclear leukocytes (PMNLs) and monocytes was demonstrated, named soluble elastin peptide receptor. Soluble elastin peptides, like K-elastin, which are liberated from elastin fibres, can be found in the sera, and they possess several biological activities such as chemotaxis. Studying the effects of elastin peptides on leukocytes, it was found that: (i) the elastin peptide stimulates the oxidative burst, the intracellular free Ca2+ elevation through a specific receptor; and (ii) in the signal transduction mechanism of this elastin peptide receptor, the phosphatidylinositol breakdown is involved. PMID- 2556300 TI - Inhibitors of arachidonic acid lipoxygenase impair the stimulation of inositol phospholipid hydrolysis by the T lymphocyte mitogen phytohaemagglutinin. AB - Piriprost and nordihydroguiaretic acid (NDGA), specific inhibitors of arachidonate lipoxygenase, inhibited phytohaemagglutinin (PHA)-stimulated breakdown of inositol lipids in human T lymphocytes. The dual inhibitors eicosatetraynoic acid (ETYA) and BW 755C, which inhibit both lipoxygenase and cyclooxygenase, also had similar actions, whereas indomethacin and acetylsalicyclic acid, which inhibit cyclooxygenase alone, did not. The effects of lipoxygenase inhibitors and dual inhibitors were reversible. These agents did not inhibit phosphatidylinositol-4,5-bisphosphate-specific phospholipase C (PIP2 PLC) in vitro. Bromophenacyl bromide, and irreversible inhibitor of phospholipase A2, also abolished PHA-stimulated inositol lipid breakdown without affecting PIP2 PLC in vitro. The results are consistent with a role for the PHA-stimulated generation of arachidonic acid and its conversion to lipoxygenase metabolites (e.g. leukotrienes and/or hydroxyeicosatetraenoic acids) as intermediate steps in the signal transduction pathway between cell-surface mitogen receptors and the stimulation of PIP2-PLC in lymphocytes. PMID- 2556301 TI - Multiple GABAA receptor alpha subunit mRNAs revealed by developmental and regional expression in rat, chicken and human brain. AB - GABAA receptor alpha subunit transcripts were detected by Northern analysis of rat, chicken and human brain mRNA using a series of 32P-labelled antisense RNA probes derived from human alpha 1 subunit cDNAs. These alpha subunit mRNAs differ in their distribution among various brain regions in the rat and at least one species is detected primarily in fetal brain. GABAA receptor alpha 1 subunit probes encoding the putative extracellular domain detect at least five alpha subunit transcripts in rat brain, whereas probes encoding the putative intracellular domain detect only two mRNAs. These data suggest the presence in brain of multiple GABAA receptor alpha subunits having homologous extracellular domains and whose expression is regionally and developmentally regulated. These alpha subunit transcripts may encode proteins that comprise GABAA isoreceptors differing in their pharmacological and physiological properties. PMID- 2556302 TI - [Dynamic changes in serum osteocalcin levels in the perinatal periods]. AB - Osteocalcin is a bone-specific protein released into the blood proportional to the rate of new born formation. It is widely accepted that the level of serum osteocalcin is a clinical marker of bone turnover. Nephrogenic cAMP is a specific indirect parameter of the biologically active parathyroid hormone. For analysis of bone metabolism during pregnancy, we measured the concentrations of osteocalcin and nephrogenic cAMP in the maternal serum during pregnancy and in the cord serum at delivery. Nephrogenic cAMP values (n mol/dl GF: mean +/- SEM) increased from the first trimester (1.5 +/- 0.21) to the term (2.11 +/- 0.11). Osteocalcin values (ng/ml: mean +/- S.D.) conversely declined from the first trimester (3.17 +/- 1.66) until the term (1.48 +/- 0.71) and acutely increased in the puerperium (5.91 +/- 2.58). These results might indicate that pregnancy induces a state of secondary hyperparathyroidism, but bone turnover is suppressed. In the cases of uncomplicated deliveries, the concentration of osteocalcin in the umbilical vein was significantly higher than that in the cord artery. This result suggests that a protein immunologically reactive to the osteocalcin antibody might be produced in the human placenta. PMID- 2556303 TI - [The role of Na+-K+ ATPase activity on blood pressure regulation and sodium metabolism in DOCA-treated rats]. AB - In this study, we attempt to clarify the role of sodium-potassium adenosine 5' triphosphatase (N+-K+ ATPase) on the regulation of systemic blood pressure and renal sodium metabolism. Male Wistar rats (n=20) were divided into four groups: 1) controls (n=5), 2) sodium loading rats (NaCl group, n=5), 3) deoxycorticosterone (DOCA group, n=5) injected rats with intact kidneys (DOCA group, n=5), 4) DOCA and ouabain-injected rats with intact kidneys (ouabain groups) and raised for six weeks. The changes in urine volume and urinary sodium were determined, and urinary excretions of norepinephrine (NE), epinephrine (E) and dopamine were measured in week six. Then blood samples were obtained from the inferior vena cava, and plasma cyclic AMP (cAMP) and cyclic GMP (cGMP) concentration were measured. At the time of sacrifice, systolic blood pressure of the ouabain group (157.3 +/- 24.8 mmHg) was significantly higher than the control group (124.5 +/- 6.9 mmHg) (p less than 0.05), but no significant differences were observed between the control group, NaCl group (123.7 +/- 7.0 mmHg), and DOCA group (128.1 +/- 4.5 mmHg). Urine volume and sodium excretion in the NaCl, DOCA and ouabain groups all increased, but the natriuretic response in the NaCl group was smaller than in either the DOCA and ouabain groups. Urinary NE in the NaCl, DOCA and ouabain groups was 1396 +/- 1025 ng/day, 640 +/- 351 ng/day, 607 +/- 177 ng/day, respectively, and NE excretions in these groups were higher than in the control group (138 +/- 104 ng/day). Urinary E in the NaCl group was higher than in the control group, but there were no significant differences among the control, DOCA and ouabain groups. Urinary dopamine in the NaCl (5723 +/- 2028 ng/day), DOCA (7661 +/- 5992 ng/day) and ouabain (4077 +/- 1984 ng/day) groups was higher than in the control group (2081 +/- 483 ng/day). Plasma cAMP in the DOCA and ouabain groups was slightly higher than in the control group, while no significant differences in plasma cAMP between the control and NaCl groups were observed. Plasma cGMP in the DOCA group was slightly higher than in either of the other three groups. These results suggest that Na+-K+ ATPase may play an important role in the maintenance of blood pressure through sodium efflux and that ouabain, Na+-K+ ATPase inhibitor, may change the renal sodium excretion and vascular responsiveness to endogenous pressor substances, leading to higher blood pressure after the administration of sodium and mineralocorticoid. PMID- 2556304 TI - [Studies on the fetal and neonatal secretion of glucagon and the development of glucagon receptors]. AB - In order to clarify the potential roles of glucagon for energy induction during the perinatal period, the developmental changes of serum glucagon concentration, the ontogenesis of hepatic glucagon receptor and glucagon-sensitive adenylate cyclase system were estimated in comparison with insulin in rats. In fetal rats on day 18 to 20 gestation, serum glucose levels and hepatic glycogen contents gradually increased as pregnancy progressed. The levels of serum glucagon and its binding to liver microsomal membranes were significantly lower than in adults, and glucagon-sensitive c-AMP production was also markedly suppressed. On day 21 of gestation, hepatic glycogen contents markedly increased to the maximal level, serum glucagon level increased, and glucagon receptors in the fetal liver were already estimated at the same level as in the adult. However, glucagon-sensitive c-AMP production was still suppressed the same as in the fetuses of earlier gestational ages. On the other hand, serum insulin levels in fetuses were higher than those in adults, and the abundant receptor could also be observed in liver microsomal membranes. After delivery, serum glucose rapidly decreased with a marked decline of hepatic glycogen contents until 5 hours in the neonatal period. Serum glucagon and its hepatic receptors were significantly increased with a gradual development of the glucagon-sensitive adenylate cyclase system. Conversely, serum insulin levels were suppressed without any remarkable change in its receptor. From these results, it is suggested that glucagon plays an important role in the neonatal adaptation mechanism, especially in the production of endogenous glucose in place of the transplacental supply from the mother, such effects of glucagon are already initiated by the induction of its receptor in target tissues in the fetus on late gestation. PMID- 2556305 TI - [A study on PP family peptide receptor: I. The distribution of peptide YY (PYY) receptor in the brain]. AB - Peptide YY (PYY) was discovered in the porcine gut by Tatemoto in 1982. PYY has recently been found in the central nervous system. This has provoked studies of PYY effects when centrally administrated. We investigated the specific binding of radioactive PYY (125I-PYY) to brain membranes in pigs and dogs. PYY chiefly bound to the hippocampus, as well as to the pituitary gland, hypothalamus, and amygdala, suggesting that PYY acts on the limbic-hypothalamic-pituitary axis. PYY binding in the brain had a high-affinity and a low-affinity component (dissociation constant, 1.39 X 10(-10) M and 3.72 X 10(-8) M, respectively). The binding sites were highly specific for PYY and neuropeptide Y (NPY), but not for pancreatic polypeptide (PP) or other peptide hormones such as cholecystokinin octapeptide, methionine enkephalin, adrenocorticotropic hormone, and thyrotropic releasing hormone. The similar affinities for PYY and NPY imply that these peptides regulate brain functions through interaction with common receptor site(s). PMID- 2556306 TI - Purification and characterization of phosphatidylinositol-specific phospholipase C from bovine spermatozoa. AB - 1. The distribution of phosphatidylinositol3, phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate hydrolysis or phosphatidylinositol specific phospholipase C (PI-PLC), activity in the bull reproductive system showed the highest specific activity in the isolated spermatozoa (SZ) followed by testis and different epididymal segments. Both the head and tail fractions of SZ were active. 2. The optimal solubilization of the enzyme from SZ was obtained with 0.2% Triton X-100 or at 0.05% detergent concentration when combined with a 60 sec sonication. The sucrose gradient centrifugation showed that PI-PLC was enriched in membrane fraction distinct from mitochondria and acrosomes. 3. The enzyme was purified by ammonium sulphate precipitation and fractionations by hydrophobic interaction chromatography, gel filtration, Con A-Sepharose affinity and chromatofocusing columns. The purified enzyme was able to hydrolyse all phosphatidylinositol substrates with optimum at pH 7.0 and activation by Ca2+, Cd2+ and Mn2+ but not phospholipids lacking the inositol residue. 4. In PAGE (8 25% gradient) the purified (aggregated) enzyme did not enter the gel. In SDS-PAGE two closely located bands were found with Mr-values of 15,000 and 18,000. Isoelectric focusing showed a wide band at pl 4.5-5.1. 5. Gel filtration resulted in a broad elution peak indicating multiple molecular forms (aggregates); the basic form had an apparent molecular weight of 100,000. The binding of the enzyme to Con A-Sepharose indicated that the enzyme is a glycoprotein. PMID- 2556307 TI - Expression of a viral gene in insulin-producing cell lines renders them susceptible to immunological destruction. AB - The gene coding for the glycoprotein D of herpes simplex virus type 1 was cloned into plasmids under the transcriptional control of the SV40 promoter-enhancer or the rat insulin 1 promoter-enhancer sequences. These plasmids were transfected into rat insulinoma cells (RINm5F) and mouse NIH/3T3 cells and the expression of glycoprotein D was examined using cell surface immunofluoresence. The rat insulin 1 promoter-enhancer sequences directed efficient expression in RINm5F cells, but not in NIH/3T3 cells. In contrast, the SV40 promoter-enhancer sequences worked well in NIH/3T3 cells, but not in RINm5F cells. Expression of glycoprotein D did not interfere with insulin production by RINm5F cells. When stable cel lines expressing glycoprotein D were exposed to anti-herpes simplex virus type 1 antibodies and complement, they were destroyed. These studies provide additional evidence that specific promoter-enhancer elements are required for efficient gene expression in certain cell types and demonstrate that the expression of foreign antigens on the surface of insulin-producing cells can lead to their immunological destruction. PMID- 2556308 TI - The Swedish childhood diabetes study III: IgM against coxsackie B viruses in newly diagnosed type 1 (insulin-dependent) diabetic children--no evidence of increased antibody frequency. AB - Sera from essentially all Swedish children aged 0-14 years with Type 1 (insulin dependent) diabetes mellitus with onset during an autumn period (October-December 1985) and a late spring period (May-June 1986) were selected. In all, 98 patients were analysed for IgM antibodies against coxsackie B virus serotypes 1 through 5 by a mu-antibody capture radio immunoassay technique. Sera from 94 referent children matched for age, sex and residential area, collected during the same period, were also analysed. During the autumn period, 10 out of 67 (15%) diabetic children were IgM positive while 14 out of 75 (19%) of the healthy referent children demonstrated positivity. During the late spring period only one out of 31 (3%) children with diabetes and two out of 19 (10%) referent children were IgM positive. In the diabetic patients, five were coxsackie B2 positive while coxsackie B1, 3, 4 and 5 were represented by one to three patients each. Eight referent children were coxsackie B4 positive, six were B3 positive and two B2 positive, while no referent children were positive against coxsackie B1 and 5. During these two periods in late 1985 and early 1986 these data demonstrate no evidence of increased antibody frequency against coxsackie B virus 1 through 5 at the onset of childhood diabetes in Sweden. PMID- 2556309 TI - Enalapril retards glomerular basement membrane thickening and albuminuria in the diabetic rat: are these effects specific for enalapril? PMID- 2556310 TI - [The relation of the constitutional test--the capacity to perceive the taste of phenylthiocarbamide--to the status of the cyclic nucleotide system]. PMID- 2556312 TI - Animal models of AIDS. AB - Animal models of AIDS are essential for understanding the pathogenesis of retrovirus-induced immune deficiency and encephalopathy and for development and testing of new therapies and vaccines. AIDS and related disorders are etiologically linked to members of the lentivirus subfamily of retroviruses; these lymphocytopathic lentiviruses are designated human immuno-deficiency virus type 1 (HIV-1) and human immuno-deficiency virus type 2 (HIV-2). The only animals susceptible to experimental HIV-1 infection are the chimpanzee, gibbon ape, and rabbit but AIDS-like disease has not yet been reported in these species. Macaques can be persistently infected with some strains of HIV-2 but no AIDS-like disease has resulted. It is not yet clear how suitable HIV-infected SCID-hu mice will be as a model for AIDS. Several subfamilies of naturally occurring cytopathic retroviruses cause immune suppression, including fatal immunodeficiency syndromes in chickens, mice, cats, and monkeys. Domestic cats suffer immunosuppression from both an onco-virus, feline leukemia virus, and a member of the lentivirus subfamily, feline immunodeficiency virus (FIV). Asian macaques are susceptible to fatal simian AIDS from a type D retrovirus, indigenous in macaques, and from a lentivirus, simian immunodeficiency virus (SIV), which is indigenous to healthy African monkeys. SIV is the animal lentivirus most closely related to HIV. Of these animal models, the lentivirus infections of cats (FIV) and macaques (SIV) appear to bear the closest similarity in their pathogenesis to HIV infection and AIDS. This review will summarize these various animal model systems for AIDS and illustrate their usefulness for antiviral therapy and vaccinology. PMID- 2556311 TI - Bacterial adaptation to oxidative stress: implications for pathogenesis and interaction with phagocytic cells. AB - During phagocytosis, phagocytic cells generate superoxide and other reactive oxygen species, which are involved in antibacterial activity. However, many bacteria possess antioxidant defenses that may explain their survival in inflammatory foci. These defenses include antioxidant enzymes such as superoxide dismutase and catalase, DNA repair systems, scavenging substrates, and competition with phagocytes for molecular oxygen. These defenses are probably coordinated, and different responses occur with different reactive oxygen species. Escherichia coli and Salmonella typhimurium mutants have allowed the demonstration of a variety of critical genes for enzymatic defense and DNA repair, as well as an oxyR regulon system. In more complex systems, the conditions found in inflammatory foci, such as decreasing glucose and the production of lactate, enhance bacterial catalase production and resistance to hydrogen peroxide. Resistance and adaptation to phagocyte-derived oxidant stress are critical aspects of bacterial pathogenesis. PMID- 2556313 TI - Recycling of tumor necrosis factor-alpha receptor in MCF-7 cells. AB - Kinetics of regulation of membrane receptors for tumor necrosis factor-alpha (TNF) in human breast adenocarcinoma MCF-7 cells was investigated. When MCF-7 cells were incubated with radioiodinated human recombinant TNF, they bound TNF specifically and accumulated it intracellularly. Preincubation of cells with native TNF up to 1 x 10(-9) M for 12 h stimulated specific binding of TNF, indicating that concentrations of membrane receptors for TNF were regulated by the ligand. Accumulation of radioactivity in cells incubated with [125I]TNF proceeded at a constant rate for up to 24 h. Kinetics of binding and internalization of TNF were similar in the presence and absence of protein synthesis for at least 1 h, suggesting that the TNF receptor was either replenished from an intracellular pool of receptors or was recycled (reutilized) during the course of TNF internalization. Data were analyzed kinetically by fitting equations of compartmental models of ligand-cell interactions with and without the term for receptor recycling. Fits were obtained only for the model with receptor recycling; absence of the term for receptor recycling resulted in physically impossible best-fit parameter values. Analysis of the best-fit parameters indicated that both internalization and recycling of the receptor were stimulated by the ligand. PMID- 2556314 TI - Cytotoxic activities of a fusion protein comprised of TGF alpha and Pseudomonas exotoxin. AB - A cDNA encoding transforming growth factor type alpha (TGF alpha) was fused to the 5' end of a gene encoding a modified form of Pseudomonas exotoxin A (PE), which is devoid of the cell recognition domain (domain Ia). The chimeric molecule, termed TGF alpha-PE40, was expressed in Escherichia coli and isolated from the periplasm or inclusion bodies depending on the construction expressed. TGF alpha-PE40 was found to be extremely cytotoxic to cells displaying epidermal growth factor (EGF) receptors. Comparison with a similar molecule in which TGF alpha was placed at the carboxyl end of PE40 demonstrated the importance of the position of the cell recognition element; TGF alpha-PE40 was found to be about 30 fold more cytotoxic to cells bearing EGF receptors than PE40-TGF alpha. In addition, TGF alpha-PE40 was shown to be extremely cytotoxic to a variety of cancer cell lines including liver, ovarian, and colon cancer cell lines, indicating high levels of EGF receptor expression in these cells. PMID- 2556315 TI - [Chemotherapy combined with Lipiodol. In vitro study of the kinetics of release of adriamycin]. AB - Intra-arterial injection of lipiodol-adriamycin mixtures are commonly used in the treatment of hepatic tumors based on the progressive release of adriamycin. This study was undertaken to assess, in vitro, the influence of mixture formulations on the adriamycin release pattern. Eight mixtures containing 10 mg of adriamycin were tested. Adriamycin was tested in solution (mixture A) in suspension (mixture B), or in emulsions with Hexabrix 320 (mixtures C to F). Ratios between Hexabrix and lipiodol volumes were 2/1, 1/1, 1/2, and 1/4 for emulsions C, D, E, and F, respectively. Emulsions G and H corresponded to emulsions E and F, with Arlacel A as emulsifying agent. All mixtures were prepared in triplicate and added with water. Samples of 200 microliters were taken from the aqueous phase after 10, 20, 30 min, 1, 2, 4, 8, 24, 48, 72, and 120 h for adriamycin dosage. Lipiodol adriamycin mixture formulation significantly influenced the release pattern of adriamycin. Three formulations (suspension, emulsions 2/1 and 1/4) induced the most progressive release of adriamycin from lipiodol. This release was dramatically retarded by the addition of an emulsifying agent. PMID- 2556316 TI - [Advanced hepatocellular carcinoma: value of systemic chemotherapy combining adriamycin, amiodarone and 5-fluorouracil]. PMID- 2556317 TI - [An extra-pancreatic site of glucagonoma: splenic metastasis of unknown pancreatic tumor or glucagonoma of an aberrant pancreas? Study of a case]. PMID- 2556318 TI - G-protein regulation of neuronal voltage-activated calcium currents. PMID- 2556319 TI - Possible mechanism of the stimulatory effect of 4-aminopyridine on toad skin. AB - 1. This study was undertaken to demonstrate the facilitatory effects of 4 aminopyridine on transmitter release in the toad skin preparation. 2. A dose dependent increase in the bioelectric parameters was found. 3. This effect was blocked by propranolol and by verapamil but not by dibenamine, and was mimicked by the calcium ionophore A-23187 but not by potassium channel blockers. 4. These results support the hypothesis that 4-aminopyridine increases presynaptic nerve terminal permeability of calcium and thus the release of noradrenaline. PMID- 2556320 TI - Inhibition constants and GABA-shifts at 2 degrees C and 37 degrees C for a spectrum of ligands acting on the benzodiazepine receptors of guinea pig hippocampus. AB - 1. Dissociation constants and Hill coefficients were determined for 13 ligands inhibiting the binding of [3H]flunitrazepam to the benzodiazepine receptors of guinea pig hippocampus at 2 degrees C and 37 degrees C. 2. The ratio of I50 in the absence of gamma-aminobutyric acid (GABA) to that in the presence of 10(-5) M GABA (the GABA-shift) was found to vary from 0.4 to 2.5. 3. There was no correlation between the pharmacological activity of the ligand and the GABA shift, or between the pharmacological activity and the magnitude of the increase in affinity of the ligand for the receptor as the temperature decreased from 37 degrees C to 2 degrees C. 4. A correlation was observed between the temperature induced affinity change and the GABA-shift at 37 degrees C. PMID- 2556321 TI - Mediation of cyclic adenosine 3',5'-monophosphate in 1-methyladenine production by starfish ovarian follicle cells. AB - Resumption of meiosis in starfish oocytes is induced by 1-methyladenine (1-MeAde) produced by ovarian follicle cells under the influence of a gonad-stimulating substance (GSS). It has also been reported that concanavalin A (Con A) and two serine proteolytic enzymes (trypsin and Pronase) can stimulate 1-MeAde production. This study was undertaken to determine if 1-MeAde production induced by these compounds is mediated through elevation of cAMP in starfish (Asterina pectinifera) follicle cells. GSS at a concentration of 0.1 mg/ml significantly stimulated 1-MeAde accumulation in extracellular medium after 1-2 hr of follicle cell incubations. GSS also caused a four- to fivefold increase in intracellular levels of cAMP. The continuous presence of GSS was required for the maintenance of elevated levels of cAMP and 1-MeAde. Basal levels of intracellular cGMP were only about 20% of those of cAMP and were not influenced by treatment with GSS. At 1 mM, 3-isobutyl-1-methylxanthine (IBMX), a potent phosphodiesterase inhibitor, stimulated both 1-MeAde and cAMP production in a concentration-dependent manner. Con A and two serine proteases also raised both cAMP and 1-MeAde production. Con A-induced (1.0 mg/ml) increases in cAMP and 1-MeAde were greater than the response to GSS (0.1 mg/ml) and were completely suppressed by treatment with alpha-methyl-D-mannoside (10 mM), a competitive inhibitor of Con A. These results strongly suggest that cAMP is a second messenger in the production of 1-MeAde by starfish ovarian follicle cells. PMID- 2556322 TI - Effects of bencyclane on normal and cevadine-modified Na channels in frog skeletal muscle. AB - The effect of 10(-5) mol/l bencyclane on the repetitive electrical activity of muscle membrane was studied with the conventional microelectrode technique. Electrical activity was induced by repetitive stimulation in normal Ringer solution (train) or by a single depolarizing current pulse in the presence of 10( 6) mol/l cevadine (volley). Bencyclane decreased, in a use-dependent manner, the maximum rates of depolarization and repolarization (Vmax+ and Vmax-, resp.) of the action potentials both of the train and the volley. The inhibition of Vmax+ and Vmax- was proportional; however, it was stronger for the volleys than for the trains. The cycle length (mean interspike interval) of the volley was increased by bencyclane; the prolongation was progressive during consecutive cycles. The dissociation of bencyclane from the Na channel was studied by applying trains of different durations with equal pulse numbers. Bencyclane at a higher concentration (5 x 10(-5) mol/l) caused a reversible tonic block: the overshoot potentials, Vmax+ and Vmax- were markedly reduced. The reduction of Vmax- was slightly stronger than that of Vmax+. Slow membrane potential oscillation (SMPO) was evoked by treating the muscle with 10(-4) mol/l of cevadine. The administration of 5 x 10(-6) mol/l bencyclane decreased the frequency of SMPO, while 10(-5) mol/l bencyclane terminated the slow oscillation activity without changing its baseline potential. The present results indicate that bencyclane induces use-dependent inhibition of Na channels in muscle, similarly as do class 1 antiarrhytnmic drugs. Inhibition was observed with both normal and cevadine modified Na channels. PMID- 2556323 TI - An inert hydrogen ion buffer for sodium current measurements in Ranvier nodes. PMID- 2556324 TI - Effect of bile salts on proton binding to human serum albumin. PMID- 2556325 TI - Transposable elements and fitness in Drosophila melanogaster. AB - Transposable elements constitute a significant fraction of the Drosophila melanogaster genome. The five families of moderately repeated transposable elements identified to date occupy dispersed and variable genomic locations, but have relatively constant copy numbers per individual. What effect to these elements have on the fitness of the individuals harboring them? Experimental evidence relating to this question is reviewed. The relevant data fall into two broad categories. The first involves the determination of the distribution of transposable elements in natural populations, by restriction mapping or in situ hybridization, and the comparison of the observed distribution with different theoretical expectations. The second approach is to study directly the effects of new transposable element-induced mutations on fitness. The P family of transposable elements is a particularly efficient mutagen, and the results of experiments in which initially P-free chromosomes are contaminated with P elements are discussed with regard to P-induced fitness mutations. PMID- 2556327 TI - Recombination of bacteriophage lambda in recD mutants of Escherichia coli. AB - RecBCD enzyme is centrally important in homologous recombination in Escherichia coli and is the source of ExoV activity. Null alleles of either the recB or the recC genes, which encode the B and C subunits, respectively, manifest no recombination and none of the nuclease functions characteristic of the holoenzyme. Loss of the D subunit, by a recD mutation, likewise results in loss of ExoV activity. However, mutants lacking the D subunit are competent for homologous recombination. We report that the distribution of exchanges along the chromosome of Red-Gam-phage lambda is strikingly altered by recD null mutations in the host. When lambda DNA replication is blocked, recombination in recD mutant strains is high near lambda's right end. In contrast, recombination in isogenic recD+ strains is approximately uniform along lambda unless the lambda chromosome contains a chi sequence. Recombination in recD mutant strains is focused toward the site of action of a type II restriction enzyme acting in vivo on lambda. The distribution of exchanges in isogenic recD+ strains is scarcely altered by the restriction enzyme (unless the phage contains an otherwise silent chi). The distribution of exchanges in recD mutants is strongly affected by lambda DNA replication. The distribution of exchanges on lambda growing in rec+ cells is not influenced by DNA replication. The exchange distribution along lambda in recD mutant cells is independent of chi in a variety of conditions. Recombination in rec+ cells is chi influenced. Recombination in recD mutants depends on recC function, occurs in strains deleted for rac prophage, and is independent of recJ, which is known to be required for lambda recombination via the RecF pathway. We entertain two models for recombination in recD mutants: (i) recombination in recD mutants may proceed via double-chain break--repair, as it does in lambda's Red pathway and E. coli's RecE pathway; (ii) the RecBC enzyme, missing its D subunit, is equivalent to the wild-type, RecBCD, enzyme after that enzyme has been activated by a chi sequence. PMID- 2556326 TI - Natural selection and ribosomal DNA in Drosophila. AB - Natural populations of Drosophila mercatorum are variable for the number of X linked 28S ribosomal genes bearing a 5-kilobase insert. A separate polymorphic X linked gene controls whether 28S repeats bearing the insert are preferentially underreplicated during the formation of polytene tissue. Female flies having at least a third of their 28S genes bearing the insert and lacking the ability to preferentially underreplicate inserted repeats display the abnormal abdomen syndrome. The syndrome is characterized by retention of juvenile abdominal cuticle into the adult, a slowdown in larval developmental time, and an increase in early female fecundity. The life history traits are expressed in nature and provide a basis for strong natural selection. The abnormal abdomen syndrome should be favored whenever the adult age structure is skewed towards young individuals, and field studies confirm this prediction. The closely related species, Drosophila hydei, also bears these inserts and appears to be subject to similar selection. However, D. mercatorum responds to this selection primarily through the allelic variation that controls preferential underreplication, whereas D. hydei responds primarily through adjustment of the proportion of inserted 28S genes. This is interpreted to mean that the evolution of a multigene family arises from the interaction of population-level and DNA-level processes. PMID- 2556328 TI - Expression and activity of a gene encoding rat cytochrome c in the yeast Saccharomyces cerevisiae. AB - A rat-processed pseudogene, which encodes normal rat cytochrome c, has been expressed in the yeast, Saccharomyces cerevisiae. The translated region of the chromosomal CYC1+ locus, which encodes yeast iso-1-cytochrome c, was replaced by the translated region of the gene encoding rat cytochrome c (CYC1-RAT), thus preserving the proper CYC1 transcription initiation and termination signals. Although the levels of transcription of the normal CYC1+ gene and the CYC1-RAT gene in yeast were equivalent, rat cytochrome c was produced at approx. 40% of the level of iso-1-cytochrome c. In addition, the specific activity in vivo was estimated to be approx. 60% that of the yeast iso-1-cytochrome c. N-terminal processing of indigenous rat cytochrome c, in which the N-terminal methionine residue is cleaved and the penultimate glycine residue is acetylated, also occurred in yeast. Methionine cleavage was complete, while acetylation proceeded to only 70% completion. Lys-72 was trimethylated to 66% completion in the rat cytochrome c produced in yeast. The near normal expression (40%) and specific activity (60%) in vivo indicates that the 40% difference in amino acid sequence is not critical for mitochondrial import, heme attachment and interactions with redox partners. PMID- 2556329 TI - Human ornithine decarboxylase(ODC)-encoding gene: cloning and expression in ODC deficient CHO cells. AB - We have cloned a full-length human ornithine decarboxylase (ODC)-encoding gene from a genomic library of human myeloma cells which overproduce ODC due to a selective gene amplification. Correct expression of the cloned gene was assessed by transfecting it into a Chinese hamster ovary (CHO) cell mutant devoid of ODC activity. Transfection with a 10-kb BamHI DNA fragment of the genomic clone, conferred ODC activity to the recipient cells and relieved them of dependence on exogenous polyamines for growth. A set of 40 transformants was isolated, eight of which were further characterized. The transfected ODC gene appeared to be hypomethylated at the cytosine residues in the sequence CpG. The transfectants were all responsive to serum stimulation, but showed different levels of ODC expression depending on both copy number and integration site of the transfected ODC gene. ODC serum induction in the transfectants was sensitive to cycloheximide and polyamine additions, and the half-life of the enzyme was very short, like that in normal CHO cells. These results suggest that the human ODC gene we transfected contains all the elements needed for normal control of ODC expression. PMID- 2556330 TI - The differential overamplification of short sequences in the mitochondrial DNA of rho- petites in Saccharomyces cerevisiae stimulates recombination. AB - It is well known that the yeast Saccharomyces cerevisiae can be affected by mutations, called 'petite colonie', which correspond to the loss of the major part of the mitochondrial DNA and the concomitant amplification of the remaining sequence, the basic repeat unit (BRU). We describe here a new phenomenon, the internal overamplification (IOA), due to the differential amplification (up to 20 fold) of short sequences within the BRU. These IOAs are very stable and stimulate the recombination. We discuss here the possible mechanisms giving rise to the appearance and maintenance of the IOAs within the BRU and their effect on the recombination process. PMID- 2556331 TI - [Carcinogenicity of zeolite-phillipsite]. AB - Phillipsite dust was intrapleurally administered to random-bred rats by 20 mg in 0.5 ml of saline three times with an interval of a month. Higher incidence of hemoblastoses has been experimentally found in exposed animals compared to the control ones. One rat developed pleural mesothelioma, two rats lung adenocarcinoma. It was concluded that phillipsite has weak carcinogenic activity. PMID- 2556332 TI - [Effect of cation flocculants on Enterobacteriaceae and Enteroviruses]. PMID- 2556333 TI - [Surgical procedures in multiple primary synchronous bilateral tumors of the lungs]. PMID- 2556334 TI - [Viral infections in the granulocytopenic patient]. PMID- 2556335 TI - The role of hydrogen cyanide and carbon monoxide in fire casualties: a prospective study. AB - Determinations of blood cyanide and carboxyhemoglobin concentrations were performed in 18 victims found dead in buildings after fires during a 2-year period. The results indicated that 50% of the victims had been exposed to toxic levels of hydrogen cyanide and 90% to toxic levels of carbon monoxide. Lethal concentrations of carbon monoxide were found in 83% of the victims. In one case a lethal blood cyanide but a non-toxic blood carboxyhemoglobin value was found. It is concluded that carbon monoxide appears to be more important than hydrogen cyanide as a toxic agent in the fire atmosphere, but cyanide poisoning without carbon monoxide poisoning may, under certain circumstances, be the cause of death in fire victims. PMID- 2556336 TI - [Analysis of 293 cases of hepatocellular carcinoma of recent ten years--I. Epidemiology, anamnesis and tumor occupying rate]. AB - Two hundred and ninety-three cases of hepatocellular carcinoma (HCC) admitted to the key hospital of the rural area of western Japan, Saga Prefectural Hospital, during 10 years since 1979 to 1988 were analysed retrospectively. The male to female ratio was 3.7 to 1 and the ratio of HBs-Ag positive to negative was 3.5 to 1. The peak of the age distribution was seventh decade. The incidence increased annually, especially in HBs-Ag negative males. Their past history of blood transfusion was considered as an etiology of HCC in 19 (9.9%). The HCC was diagnosed approximately 10 years earlier in habitual drinkers than non-habitual drinkers of both positive and negative HBs-Ag. The incidence of small tumors is associated with the past history of chronic liver disease, negativity of HBs-Ag, lack of symptoms and old age. PMID- 2556337 TI - [Analysis of 293 cases of hepatocellular carcinoma of recent ten years--II. Diagnosis, therapy and prognosis]. AB - Two hundred and ninety-three cases of hepatocellular carcinoma (HCC) of recent ten years during 1979 and 1988 were analysed on its diagnosis, therapy and prognosis. Angiography was the most useful diagnostic method for the first 5 years. During recent 5 years and for the tumor with size less than 2 cm, however, ultrasonography was the best method. 24.7% cases showed negative AFP and only 42.5% cases showed the level over 400 ng/ml. According to the tumor occupying rate at the diagnosis, the number of cases of E1 had increased annually, the other cases being unchanged. 88.6% cases had liver cirrhosis microscopically as being reported previously. As a therapy TAE was most frequently performed, followed by operation, arterial infusion of antitumor drugs et al. The prognosis of the patients depended on the tumor occupying-rate. For the case of E1, the prognosis was the best by the therapy of ethanol injection therapy, operation and TAE. For the cases with tumor of larger than E1, frequent TAE was the best. From these results we concluded that we must make efforts to find small tumors to get the good prognosis. PMID- 2556338 TI - Motility and fertilizing ability of rat epididymal spermatozoa washed by a continuous gradient of Percoll. AB - Removal of epididymal fluids from epididymal sperm suspension is an important step for the study of sperm motility, capacitation, and the acrosome reaction. The technique of washing should minimize damage to viable spermatozoa but at the same time efficiently remove debris, non-sperm cells, and biological fluids. We examined sperm motility and fertilizability in vitro of rat epididymal spermatozoa after washing with Percoll continuous gradient. Nine milliliters (ml) of 50% N-2-hydroxyethylpiperazine-N1-2-ethanesulfonic acid (HEPES) buffered Percoll solution was centrifuged at 20,000 g for 45 minutes to form a continuous gradient. One hundred to 300 microliters of sperm suspension was loaded onto the surface of the gradient and centrifuged at 150 g and 1,500 g for 10 minutes. Two main layers of spermatozoa were formed, one of high (lower layer) and one of low (upper layer) motility. At centrifugation 1,500 g, the sperm density and motility in the lower layer were greater than at 150 g. Spermatozoa from both layers at 150 g and at 1,500 g were diluted with modified Krebs-Ringer's bicarbonate solution (mKRB) and preincubated for 5 hours. Superovulated eggs collected from 21-25-day-old Wistar strain immature rats were introduced into the preincubated sperm suspension for insemination and fixed 5-5.5 hours later for observation of fertilization. Spermatozoa from both layers, 150 g and 1,500 g, showed the same fertilizability in vitro as control spermatozoa. From these results we conclude that Percoll gradients can be used for washing rat epididymal sperm for the study of sperm physiology including fertilization. PMID- 2556339 TI - Acrosomal function of human spermatozoa with normal and abnormal head morphology. AB - In previous studies we have shown differences in the function of morphologically normal and abnormal sperm by evaluating their flagellar movements and swimming trajectories. In this study we have compared the capability of morphologically normal and abnormal human sperm to undergo an acrosome reaction after incubation with human follicular fluid. Semen samples were studied from 6 research donors and 21 semen evaluation patients. All men had normal semen by clinical criteria. Semen was prepared either by a two-step Percoll gradient centrifugation or the sperm were diluted, washed, and centrifuged three times. Sperm suspensions were incubated for 24 hours in a modified Tyrode's medium, containing 2.6% bovine serum albumin, prior to dilution with human follicular fluid. The percentage of acrosome reactions among viable sperm was assessed after 15 minutes using the supravital Hoescht stain and fluoresceinated pea lectin. Sperm head size was measured with an ocular micrometer and normal values were defined as length 3-5 microns and width 2-3 microns. At least 25 viable normal sperm, and 25 viable abnormal sperm were analyzed for acrosome reactions on each slide. With Percoll separation the percentage of acrosome reactions (mean +/- sem) for normal sperm was 38 +/- 3% vs. 22 +/- 2% for abnormal sperm (P less than 0.005). After washing, the comparable values were 12 +/- 1% vs. 5 +/- 1% (P less than 0.005). The incidence of spontaneous acrosome reactions (24 hours of incubation, no follicular fluid) was also higher for normal sperm than abnormal sperm (9 +/- 1% vs. 4 +/- 1%, P less than 0.01). These data demonstrate an association between normal sperm morphology and acrosomal function. PMID- 2556340 TI - Oxytocin does not modify glucagon- or calcitonin-induced ACTH-cortisol rise in humans. PMID- 2556341 TI - Enzyme polymorphism and clinical variability of diseases: a study of diabetes mellitus. AB - We investigated possible relations among four common neonatal manifestations of diabetic pregnancy (macrosomia, hypoglycemia, hypocalcemia, jaundice) and four enzyme polymorphisms (PGM1, ADA, AK1, ACP1 in a sample of infants born of diabetic mothers. The pattern of associations observed between the two sets of variables is consistent with known differences in enzymatic activity within phenotypes of each system, suggesting that low enzymatic activity may have unfavorable effects on fetal development and on adaptability of the neonate to the extrauterine environment, Some of the polymorphic enzymes studied influence fetal growth in normal pregnancy as well. Analysis of relations between genetic polymorphisms and the clinical pattern of common diseases may provide a better understanding of the genetic basis of the clinical variability of diseases within and between human populations. PMID- 2556342 TI - Primary malignant fibrous histiocytoma of the pancreas. AB - A patient with a large tumor of the head and body of the pancreas had a Whipple procedure performed after the intraoperative diagnosis of "mesenchymal tumor" was made. The final histopathologic diagnosis was malignant fibrous histiocytoma arising in the pancreas. The patient died of postoperative complications, and no metastatic disease was found at autopsy. This is the first detailed description of a primary pancreatic malignant fibrous histiocytoma. PMID- 2556343 TI - CpG islands surround a DNA segment located between translocation breakpoints associated with genitourinary dysplasia and aniridia. AB - We have isolated a DNA segment absent from all the constitutionally deleted chromosomes 11 of our patients with Wilms tumor. This marker separates two balanced translocations that break in band 11p13: the distal one associated with aniridia (AN2), and the proximal one with genitourinary dysplasia (GUD). The GUD breakpoint maps within the smallest region of overlap (SRO) for the Wilms tumor (WT) gene locus, thus strengthening the previous suggestion of an association between Wilms tumor and other abnormalities of the genitourinary system. The 11p13 translocation breakpoint associated with T-cell acute lymphatic leukemia (T ALL) is centromeric to the SRO and separated from the WT locus by at least one known gene. This region of the human genome (11p13) is rich in CpG islands that potentially identify genes, some of which may be involved in the various phenotypes associated with the WAGR syndrome. This is consistent with the proposition that the majority of human genes are in G-negative bands. PMID- 2556344 TI - The gene for human muscle-specific phosphoglycerate mutase, PGAM2, mapped to chromosome 7 by polymerase chain reaction. AB - The gene for the human muscle-specific phosphoglycerate mutase, PGAM2, has been mapped to chromosome 7 using the polymerase chain reaction to specifically amplify the human PGAM2 sequence in rodent/human hybrid DNA. This assignment shows that the PGAM2 is not syntenic with the nonmuscle form of PGAM1 (B) which has been located to chromosome 10. PMID- 2556345 TI - Differential sensitivity to cAMP among human cord and maternal/adult peripheral lymphocytes discloses differences between PHA- and OKT3-induced activation pathways. AB - Previous studies have demonstrated that human cord blood lymphocytes are resistant to the anti-proliferative action of prostaglandin E2 (PGE2) in phytohaemagglutinin (PHA)-induced mitogenesis, whereas the same cells activated by OKT3 are at least as sensitive to PGE2 as the corresponding cells from their mothers or other adults. In the present investigation it was found that: (i) PHA induced proliferation of cord peripheral blood mononuclear leucocytes (PBML) is less sensitive to inhibition by forskolin--a direct activator of adenylate cyclase (AC)--and dibutyryl cAMP--a permeant cAMP analogue--than the proliferation of the corresponding maternal cells; (ii) OKT3-induced proliferation of cord as well as maternal PBML is highly sensitive to inhibition by forskolin and dibutyryl cAMP; (iii) cord PBML show an overall lower rate of AC activity compared with maternal PBML, in response to PGE2 and other autacoids as well as to receptor-independent stimulation; (iv) cord PBML also display a significantly lower rate of degradation of cAMP through cAMP-phosphodiesterase compared with maternal cells; (v) unbroken cord and maternal PBML show comparable rates of cAMP accumulation after stimulation with PGE2. The results suggest a lower sensitivity to the effect of cAMP in cord compared with maternal/adult lymphocytes in PHA-induced proliferation. Moreover, the data illustrate differences between PHA- and OKT3-mediated activation pathways, as well as differences in cell regulation mechanisms between cord and maternal PBML. PMID- 2556347 TI - Sex chromatin in female breast tumour. AB - The incidence of sex chromatin in tumour cell nuclei was estimated in twenty five cases each of carcinoma and fibroadenoma breast using haematoxylin and eosin, thionine, aceto-orcein and Papanicolaou's stains on imprint-smears. The sex chromatin incidence in fibroadenoma was found similar to that of normal breast tissue but the incidence markedly decreased in carcinoma breast. Undifferentiated tumours with histologic grade III, nuclear grade I, no lymphocytic infiltrate and involvement of overlying skin, nipple and areola were more often associated with low sex chromatin incidence. No relationship was observed between sex chromatin incidence and tumour size or axillary lymph node metastasis. PMID- 2556348 TI - Melanotic neuroectodermal tumor of infancy. PMID- 2556346 TI - Fc receptor density, MHC antigen expression and superoxide production are increased in interferon-gamma-treated microglia isolated from adult rat brain. AB - Treatment of microglia isolated from adult rat brain with interferon-gamma (IFN gamma) at a concentration of 1 U/ml resulted in enhanced expression of Fc receptors and major histocompatibility complex (MHC) antigens and increased production of superoxide anions. Neonatal microglia and peritoneal macrophages, isolated and cultured in the same manner, displayed functional properties very similar to those of adult microglia, indicating a common origin for different macrophage populations. The Fc binding capacity of microglia was found to be significantly greater than that of peritoneal cells, thus underlining the potential role of microglia in immune-mediated demyelination. PMID- 2556349 TI - [In vitro scanning electron microscope study. The role of a root surface polishing instrument in periodontal surgery demonstrated in a pocket area]. PMID- 2556350 TI - Anesthesia and chemical second messenger generation in the adrenergic nervous system. PMID- 2556351 TI - Presynaptic adrenergic effects of anesthetics. PMID- 2556352 TI - Effect of intravenously administered lipoxygenase metabolites on rat tracheal mucous gel layer thickness. AB - The effect of intravenous injections of 5-, 12- and 15-hydroxyeicosatetraenoic acids (HETE), leukotrienes D4 and E4 (LTD4, LTE4) on tracheal mucous gel layer (TMGL) thickness was assessed in rats. When administered in doses ranging from 0.03 pg to 33 ng per rat, the lipoxygenase metabolites produced significant increases in TMGL thickness. The order of potency of the metabolites was 15-HETE greater than 12-HETE greater than or equal to 5-HETE greater than LTD4 greater than or equal to LTE4. Imidazole (31.6 mg/kg), intravenously, significantly decreased this response. These findings suggest that the mono-HETEs, especially 15-HETE, may be important modulators of airway mucus in the rat. PMID- 2556353 TI - Herpes simplex virus stromal keratitis is not titer-dependent and does not correlate with neurovirulence. AB - We developed a murine model of ocular herpes simplex virus (HSV) disease which is particularly suited for testing stromal keratitis because most animals show some evidence of infection. Using this model, we characterized the ocular disease patterns caused by ten recent low-passage clinical isolates of HSV-1, as well as those caused by the established laboratory strains HSV-1 KOS and HSV-2 333. Viral strains were evaluated for their ability to cause stromal keratitis, blepharitis, vascularization of the cornea, and mortality. The model was not useful for scoring epithelial keratitis. The ocular disease caused by the recent isolates ranged from very mild disease to severe stromal keratitis. Some of the recent isolates caused disease as severe as the two laboratory strains. A comparison of the virulence characteristics expressed by various HSV strains indicated that the ability to cause stromal disease was correlated with vascularization of the cornea (correlation coefficient = 0.797, P less than 0.001) and was not correlated with the neurovirulence of the strains (correlation coefficient 0.045, P greater than 0.05). The severity of stromal keratitis was not dependent on the amount of inoculum over the range tested and a strain causing severe stromal keratitis caused severe ocular disease even when mixed with a nonstromal strain at ratios of 10:1, 100:1, and 1000:1. PMID- 2556355 TI - Effect of glucocorticoids and limiting nursing on the carbohydrate digestive capacity and growth rate of piglets. AB - The influence of glucocorticoid administration and limited nursing on piglet carbohydrase enzyme development and subsequent growth was examined in three experiments using 371 piglets. Treatments in the first two experiments were formed by the factorial arrangement of hydrocortisone (-HYD or +HYD) and limited nursing (-LN or +LN) imposed form d 14 to weaning (d 28). Hydrocortisone was replaced by adrenocorticotropic hormone (ACTH) in the third experiment. Growth rates were severely depressed by HYD (P less than .01), LN (P less than .001) and to a lesser extent (P less than .06) by ACTH during the last 2 wk of lactation. During the first 14 d postweaning, piglets continued to grow more slowly following HYD treatment (P less than .01), whereas LN piglets grew more rapidly than those allowed to suckle normally. Although piglets were smaller at weaning after HYD injection (P less than .01), relative weights of liver, pancreas and small intestine were increased (P less than .05). Only adrenal weights were increased by ACTH (P less than .09). Pancreatic and intestinal amylase activities were increased two- to three-fold by HYD injection (P less than .05) but were unaffected by ACTH or LN (P greater than .10). Sucrase and maltase activity increased linearly with age (P less than .001). This rate of increase was numerically enhanced by glucocorticoid treatment and LN. The normal decrease in lactase activity was accelerated by LN and HYD injection, with the greatest depression caused by the combination of LN and either HYD or ACTH administration (P less than .05). Glucocorticoid administration to nursing piglets can evoke premature elevation of the carbohydrase enzymes necessary for initiating the hydrolysis of starch. PMID- 2556354 TI - Methods of microphotometric assay of succinate dehydrogenase and cytochrome c oxidase activities for use on human skeletal muscle. AB - Microphotometric assay media for the measurement of succinate dehydrogenase (SDH) and cytochrome oxidase activities in sections of human skeletal muscle have been developed. The optimal constitution of these media was determined experimentally. Factors investigated include the effects of substrate concentration, pH, use of different electron acceptors and electron donors, influence of intermediate electron carriers and tissue-stabilizing agents, effects of inhibitors, the extent of endogenous and non-specific reactions and the linearity of the reactions during the time course of the assays. Optimal assay media (SDH) contained 130 mM succinate, 1.5 mM Nitro Blue tetrazolium, 0.2 mM phenazine methosulphate and 1.0 mM sodium azide in 0.1 m phosphate buffer, pH7.0. Cytochrome oxidase was optimally assayed in media containing 4 mM diaminobenzidine and 100 microns cytochrome c. Reactions in individual muscle fibers were found to be linear for incubation times up to 10 min in SDH assays and for more than 15 min in cytochrome oxidase determinations. Some potential uses of these microphotometric assays in the investigation of human metabolic muscle disorders are discussed. PMID- 2556356 TI - Response of digestive carbohydrases and growth to graded doses and administration frequency of hydrocortisone and adrenocorticotropic hormone in nursing piglets. AB - Graded levels of hydrocortisone 21-acetate (HYD) (0, 18, 16 and 24 mg/kg BW) were injected into nursing piglets every other day (Exp. 1) or 24 mg of HYD/kg BW was administered 0, 2, 4 or 6 times during the treatment period (12 d) with equal time (6 d, 3 d or 2 d) between subsequent injections (Exp. 2). Adrenocorticotropic hormone (ACTH) was injected to provide 0, 5, 10 or 15 IU/kg BW (Exp. 3), or 15 IU ACTH/kg BW was injected 0, 1, 2 or 3 times (Exp. 4). The injection treatment periods were from d 14 to d 26 postpartum. Pancreatic and intestinal amylase activity was maximized by the highest dosage of HYD (24 mg) and ACTH (15 IU) when given at 2- or 4-d intervals, respectively (P less than .10). However, four injections of HYD administered 3 d apart optimized the activity of this enzyme in Exp. 2 (P less than .05). Intestinal sucrase and maltase were unresponsive to ACTH regardless of dosage or injection frequency (P greater than .10). The response of these two enzymes to HYD was inconsistent. Maltase activity was elevated (P less than .10) by the two most frequent injection treatments, and sucrase activity was simultaneously depressed. Lactase activity tended (P less than .15) to be depressed by the highest treatment level in all four experiments. Both dosage and frequency methods of increasing HYD administration resulted in hepatic and pancreatic hypertrophy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556358 TI - Topoisomerase II as a target of antileukemia drugs: a review of controversial areas. PMID- 2556357 TI - Temporal changes in carbohydrate digestive capacity and growth rate of piglets in response to glucocorticoid administration and weaning age. AB - Two experiments were conducted that demonstrated that a single injection of hydrocortisone 21-acetate (HYD, 25 mg/kg BW) administered to 6-d-old nursing piglets resulted in a twofold elevation (P less than .02) of pancreatic amylase within 2 d; activity was unaffected by an injection of 15 IU adrenocorticotropic hormone (ACTH)/kg BW (P greater than .20). Intestinal sucrase and maltase activity tended to be elevated (P less than .20) 2 and 4 d postinjection with HYD but returned to normal (uninjected) levels by 14 d of age. The normal decline of intestinal lactase activity was delayed by at least 4 d in response to both hormones (P less than .10). Organ weights were not affected by either hormone. In a separate experiment, postweaning mortality was reduced (12 vs 27%) and growth rate was substantially improved by administration of HYD to piglets 4 and 2 d prior to weaning at 14 d of age. Hydrocortisone resulted in a faster rate of gain the 1st wk postweaning for pigs weaned at 21 or 28 d. Subsequent gain by control and HYD piglets weaned on d 21 was similar, but HYD subsequently impaired growth rate of piglets weaned at 28 d of age. Growth rates of control and ACTH piglets were similar at each postweaning period regardless of weaning age (weaning age [lin.] x week postweaning [quad.] x treatment, P less than .07). This differential treatment response of daily gain may be due in part to effects on feed intake (weaning age [lin.] x week postweaning [lin.] x treatment, P less than .10). We conclude that a single injection of HYD to 6-d-old piglets precociously induces pancreatic amylase and that the sensitivity of piglets to HYD is age-dependent. PMID- 2556359 TI - Rearrangement and expression of beta-T-cell receptor and immunoglobulin genes in established Ph1 chronic myelogenous leukemia cell lines. AB - We have determined the arrangement and expression of immunoglobulin (Ig) and beta T-cell receptor (TCR) genes in six established Philadelphia chromosome-positive (Ph1) chronic myelogenous leukemia (CML) cell lines, and correlated these results with their phenotypic characteristics. Three cell lines with nonlymphoid characteristics, EM2, EM3, and K562, did not demonstrate rearrangement or expression of Ig or beta-TCR genes. A new cell line, MB, with a mature B-cell phenotype recently established in our laboratory, contained light and heavy chain immunoglobulin gene rearrangements and expressed mature Ig RNA. In a cell line with an early lymphoid phenotype, BV173, this analysis showed rearrangement of Ig heavy chain and beta-TCR genes, unrearranged Ig light chain DNA, and expression of only an immature beta-TCR transcript. This line provides evidence for T-cell lineage involvement in Ph1 CML. One cell line without markers of any cell type, KCL-22, demonstrated rearranged, unexpressed Ig heavy chain genes, suggesting these cells are at the very earliest stages of lymphoid differentiation. These lines should provide valuable tools to dissect the molecular biology of differentiation in CML and in early lymphocytes. PMID- 2556360 TI - 5-nucleotidase activity in cultured cell lines. Effect of different assay conditions and correlation with cell proliferation. AB - The 5-AMPase activity of the ectoenzyme 5-nucleotidase has been measured in a variety of cell lines, using intact cells. Human cell types showed two orders of magnitude higher enzyme activity than mouse cell lines. The ectoenzyme is inhibited by adenosine 5-(alpha, beta-methylene) diphosphate and Concanavalin A. A different extent of 5-nucleotidase lectin inhibition was observed in the studied cell lines, suggesting that the corresponding ectoenzymes are glycoproteins with a different type or degree, or both, of glycosylation. The 5 nucleotidase activity increased during subculture and decreased after cell transformation. Generally, the 5-nucleotidase activity was two- to five-fold higher in monolayer than in suspension cell culture. A relation between cell growth and 5-AMPase activity was also observed. Enzyme activity increased at the end of the lag phase (glioblastoma cells) or during the exponential phase (the other two cell lines). After confluence, the activity decreased to the initial or even lower range of activity. Observed activity variations with cell proliferation correlate with modifications of 5-AMPase activity during subculture. PMID- 2556361 TI - Liquid chromatographic determination of vitamin D in infant formula. AB - A method is described for the determination of vitamins D2 + D3 in milk- and soy based infant formula. Vitamins D2 and D3 are extracted from the saponified sample and converted to isotachysterols with acidified butanol. Reverse-phase liquid chromatography (LC) is used to remove interferences, and total vitamin D is quantitated using normal-phase LC. Recoveries of spiked samples averaged 97.6% for milk-based infant formula, and 98.8% for soy-based infant formula. This method quantitates vitamin D2 + D3 in infant formulas containing as low as 40 IU/qt when prepared according to label direction. PMID- 2556362 TI - Bronchogenic carcinoma. A study of 109 cases. AB - The clinical profile of 109 cases of bronchogenic carcinoma has been studied. Definite histopathological typing of malignancy was possible in 61 patients, squamous cell carcinoma being the commonest tumour in 27 followed by small cell carcinoma in 16, adenocarcinoma in 11 and large cell carcinoma in 7. Another 23 patients showed changes suggestive of malignancy on histological/cytological examination. Definitive diagnosis was obtained in half of the patients by bronchial biopsy, in 16% by bronchial aspiration fluid cytology, in 13% by pleural biopsy, in 11% by lymph node biopsy and in others by pleural fluid cytology, lung biopsy and skin biopsy. Commonest radiological involvement among different cell types was hilar involvement in small cell carcinoma in 62%, evidence of bronchial obstruction (collapse/consolidation) in squamous cell carcinoma in 56%, peripheral mass in adenocarcinoma in 54% and cavitation in a mass in squamous cell carcinoma in 15% cases. PMID- 2556363 TI - Evolutionary perspectives on multiresistance beta-lactamase transposons. AB - A series of intragenic DNA probes, encoding the major part of the transposase resolvase and inverted repeats of transposons Tn3, Tn21, and Tn2501, were used in hybridization assays for homologous DNA sequences in 18 transposons studied. The tnpA and tnpR probes detected extensive homology with Tn3-like and Tn21-like elements for 11 transposons. This high degree of homology was confirmed with the 38- and 48-base-pair inverted-repeat oligonucleotide probes of Tn3, Tn21, and Tn2501. The Southern-type gel hybridization experiments localized the tnpA homologous sequences on the physical DNA maps constructed. The genetic and physical maps of the transposons were compared, as were their nucleic acid sequence homologies. These comparisons suggested a subfamily of mobile elements distinct from but related to the Tn21 group. Based on these results, an evolutionary model is proposed and a pedigree is presented for the genesis of multiresistance beta-lactamase transposons. PMID- 2556364 TI - Isolation, characterization, and inactivation of the APA1 gene encoding yeast diadenosine 5',5'''-P1,P4-tetraphosphate phosphorylase. AB - The gene encoding diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A) phosphorylase from yeast was isolated from a lambda gt11 library. The DNA sequence of the coding region was determined, and more than 90% of the deduced amino acid sequence was confirmed by peptide sequencing. The Ap4A phosphorylase gene (APA1) is unique in the yeast genome. Disruption experiments with this gene, first, supported the conclusion that, in vivo, Ap4A phosphorylase catabolizes the Ap4N nucleotides (where N is A, C, G, or U) and second, revealed the occurrence of a second Ap4A phosphorylase activity in yeast cells. Finally, evidence is provided that the APA1 gene product is responsible for most of the ADP sulfurylase activity in yeast extracts. PMID- 2556365 TI - Transcription events in the origin of replication of plasmid pSC101. AB - Insertion mutations were isolated in the origin fragment of the plasmid pSC101 after random cleavage with DNase I. The replication properties of the resulting plasmids confirmed previous findings and extended the characterization of the essential regions. Using these plasmids, we analyzed by various methods the transcription events in the pSC101 origin. In addition to the mRNA of repA, a gene coding for the self-regulated RepA protein which is essential for replication of the plasmid, we characterized a transcript, which we called RNA Y, that runs in the opposite direction and that starts in the middle of the second repeated sequence in the origin region. Like the self-regulated repA mRNA, RNA Y is weakly expressed. It does not code for a complete protein within the origin fragment but may do so in the wild-type plasmid. We also found indications for one or, possibly, two small RNA species, called RNA X, which run in the same direction as RepA and which are partially complementary to RNA Y. We postulate that RNA Y and, possibly, RNA X are implicated in the initiation of replication of pSC101. PMID- 2556366 TI - Biochemical and genetic analyses of acetoin catabolism in Alcaligenes eutrophus. AB - In genetic studies on the catabolism of acetoin in Alcaligenes eutrophus, we used Tn5::mob-induced mutants which were impaired in the utilization of acetoin as the sole carbon source for growth. The transposon-harboring EcoRI restriction fragments from 17 acetoin-negative and slow-growing mutants (class 2a) and from six pleiotropic mutants of A. eutorphus, which were acetoin-negative and did not grow chemolithoautotrophically (class 2b), were cloned from pHC79 gene banks. The insertions of Tn5 were mapped on four different chromosomal EcoRI restriction fragments (A, C, D, and E) in class 2a mutants. The native DNA fragments were cloned from a lambda L47 or from a cosmid gene bank. Evidence is provided that fragments A (21 kilobase pairs [kb]) and C (7.7 kb) are closely linked in the genome; the insertions of Tn5 covered a region of approximately 5 kb. Physiological experiments revealed that this region encodes for acetoin:dichlorophenol-indophenol oxidoreductase, a fast-migrating protein, and probably for one additional protein that is as yet unknown. In mutants which were not completely impaired in growth on acetoin but which grew much slower and after a prolonged lag phase, fragments D (7.2 kb) and E (8.1 kb) were inactivated by insertion of Tn5::mob. No structural gene could be assigned to the D or E fragments. In class 2b mutants, insertions of Tn5 were mapped on fragment B (11.3 kb). This fragment complemented pleiotropic hno mutants in trans; these mutants were impaired in the formation of a rpoN-like protein. The expression of the gene cluster on fragments A and C seemed to be rpoN dependent. PMID- 2556367 TI - Gyrase inhibitors can increase gyrA expression and DNA supercoiling. AB - Treatment of bacterial cells with inhibitors of gyrase at high concentration leads to relaxation of DNA supercoils, presumably through interference with the supercoiling activity of gyrase. Under certain conditions, however, the inhibitors can also increase supercoiling. In the case of coumermycin A1, this increase occurs at low drug concentrations. Oxolinic acid increases supercoiling in a partially resistant mutant. We found that increases in chromosomal DNA supercoiling, which were blocked by treatment with chloramphenicol, were accompanied by an increased expression rate of gyrA. This result is consistent with gyrase being responsible for the increase in supercoiling. In wild-type cells, increases in gyrA expression were transient, suggesting that when supercoiling reaches sufficiently high levels, gyrase expression declines. Oxolinic acid studies carried out with a delta topA strain showed that drug treatment also increased plasmid supercoiling. The levels of supercoiling and topoisomer heterogeneity were much higher when the plasmid contained one of several promoters fused to galK. Since oxolinic acid causes an increase in gyrA expression, it appears that gyrase levels may be important in transcription mediated changes in supercoiling even when topoisomerase I is absent. PMID- 2556368 TI - Phosphate regulon in members of the family Enterobacteriaceae: comparison of the phoB-phoR operons of Escherichia coli, Shigella dysenteriae, and Klebsiella pneumoniae. AB - The structure and function of the phoB and phoR genes of Shigella dysenteriae strains and Klebsiella pneumoniae, which are involved in regulation of the phosphate regulon, were analyzed. Complementation tests among the genes of Escherichia coli, S. dysenteriae strains, and K. pneumoniae for production of alkaline phosphatase indicate that S. dysenteriae serotype 2 and serotype 3 strains and K. pneumoniae are phoA+ phoB+ phoR+ but S. dysenteriae Sh and serotype 1 strains are phoA phoB+ phoR. Nucleotide sequences of phoB and phoR of S. dysenteriae Sh and K. pneumoniae are highly homologous to those of E. coli, except for a single base insertion found in phoR of S. dysenteriae Sh. PMID- 2556369 TI - Introduction of transposon Tn916 DNA into Haemophilus influenzae and Haemophilus parainfluenzae. AB - Enterococcus (Streptococcus) faecalis transposon Tn916 was introduced into Haemophilus influenzae Rd and Haemophilus parainfluenzae by transformation and demonstrated to transpose efficiently. Haemophilus transformants resistant to tetracycline were observed at a frequency of approximately 3 x 10(2) to 5 x 10(3)/micrograms of either pAM120 (pGL101::Tn916) or pAM180 (pAM81::Tn916) plasmid DNAs, which are incapable of autonomous replication in this host. Restriction enzyme analysis and Southern blot hybridization revealed that (i) Tn916 integrates into many different sites in the H. influenzae and H. parainfluenzae genomes; (ii) only the 16.4-kilobase-pair Tn916 DNA integrates, and no vector DNA was detected; and (iii) the Tetr phenotype was stable in the absence of selective pressure. Second-generation Tn916 transformants occurred at the high frequency of chromosomal markers and retained their original chromosomal locations. Similar results were obtained with H. influenzae Rd BC200 rec-1 as the recipient strain, which suggests host rec functions are not required in Tn916 integrative transposition. Transposition with Tn916 is an important procedure for mutagenesis of Haemophilus species. PMID- 2556370 TI - Cyclic diguanylic acid and cellulose synthesis in Agrobacterium tumefaciens. AB - The occurrence of the novel regulatory nucleotide bis(3',5')-cyclic diguanylic acid (c-di-GMP) and its relation to cellulose biogenesis in the plant pathogen Agrobacterium tumefaciens was studied. c-di-GMP was detected in acid extracts of 32P-labeled cells grown in various media, and an enzyme responsible for its formation from GTP was found to be present in cell-free preparations. Cellulose synthesis in vivo was quantitatively assessed with [14C]glucose as a tracer. The organism produced cellulose during growth in the absence of plant cells, and this capacity was retained in resting cells. Synthesis of a cellulosic product from UDP-glucose in vitro with membrane preparations was markedly stimulated by c-di GMP and its precursor GTP and was further enhanced by Ca2+. The calcium effect was attributed to inhibition of a c-di-GMP-degrading enzyme shown to be present in the cellulose synthase-containing membranes. PMID- 2556371 TI - Hydrolysis of bis(5'-nucleosidyl) polyphosphates by Escherichia coli 5' nucleotidase. AB - Two enzymatic activities that split diadenosine triphosphate have been reported in Escherichia coli: a specific Mg-dependent bis(5'-adenosyl) triphosphatase (EC 3.6.1.29) and the bis(5'-adenosyl) tetraphosphatase (EC 3.6.1.41). In addition to the activities of these two enzymes, a different enzyme activity that hydrolyzes dinucleoside polyphosphates is described. After purification and study of its molecular and kinetic properties, we concluded that it corresponded to the 5' nucleotidase (EC 3.1.3.5) that has been described in E. coli. The enzyme was purified from sonic extracts and osmotic shock fluid. From sonic extracts, two isoforms were isolated by chromatography on ion-exchange Mono Q columns; they had a molecular mass of about 100 kilodaltons (kDa). From the osmotic shock fluid, a unique form of 52 kDa was recovered. Mild heating transformed the 100-kDa isoform to a 52-kDa form, with an increase in activity of about threefold. The existence of a 5'-nucleotidase inhibitor described previously, which associates with the enzyme and is not liberated in the osmotic shock fluid, may have been responsible for these results. The kinetic properties and substrate specificities of both forms (52 and 100 kDa) were almost identical. The enzyme, which is known to hydrolyze AMP and uridine-(5')-diphospho-(1)-alpha-D-glucose, but not adenosine (5')-diphospho-(1)-alpha-D-glucose, was also able to split adenosine-(5') diphospho-(5)-beta-D-ribose, ribose-5-phosphate, and dinucleoside polyphosphates [diadenosine 5',5'''-P1,P2-diphosphate,diadenosine 5',5'''-P1,P3-triphosphate, diadenosine 5',5'''-P1,P4-tetraphosphate, and bis(5'-guanosyl) triphosphate]. The effects of divalent cations and pH on the rate of the reaction with different substrates were studied. PMID- 2556372 TI - Parathion hydrolase specified by the Flavobacterium opd gene: relationship between the gene and protein. AB - The sequence of a 1,693-base-pair plasmid DNA fragment from Flavobacterium sp. strain ATCC 27551 containing the parathion hydrolase gene (opd) was determined. Within this sequence, there is only one open reading frame large enough to encode the 35,000-dalton membrane-associated hydrolase protein purified from Flavobacterium extracts. Amino-terminal sequence analysis of the purified Flavobacterium hydrolase demonstrated that serine is the amino-terminal residue of the hydrolase protein. The amino-terminal serine corresponds to a TCG codon located 87 base pairs downstream of the presumptive ATG initiation codon in the nucleotide sequence. The amino acid composition of the purified protein agrees well with that predicted from the nucleotide sequence, using serine as the amino terminal residue. These data suggest that the parathion hydrolase protein is processed at its amino terminus in Flavobacterium sp. Construction in Escherichia coli of a lacZ-opd gene fusion in which the first 33 amino-terminal residues of opd were replaced by the first 5 residues of lacZ resulted in the production of an active hydrolase identical in molecular mass to the hydrolase isolated from Flavobacterium sp. E. coli cells containing the lacZ-opd fusion showed higher levels of hydrolase activity than did cells containing the parent plasmid. PMID- 2556373 TI - Nucleotide sequences and operon structure of plasmid-borne genes mediating uptake and utilization of raffinose in Escherichia coli. AB - The plasmid-borne raf operon encodes functions required for inducible uptake and utilization of raffinose by Escherichia coli. Raf functions include active transport (Raf permease), alpha-galactosidase, and sucrose hydrolase, which are negatively controlled by the Raf repressor. We have defined the order and extent of the three structural genes, rafA, rafB, and rafD; these are contained in a 5,284-base-pair nucleotide sequence. By comparisons of derived primary structures with known subunit molecular weights and an N-terminal peptide sequence, rafA was assigned to alpha-galactosidase (708 amino acids), rafB was assigned to Raf permease (425 amino acids), and rafD was assigned to sucrose hydrolase (476 amino acids). Transcription was shown to initiate 13 nucleotides upstream of rafA; a putative promoter, a ribosome-binding site, and a transcription termination signal were identified. Striking similarities between Raf permease and lacY encoded lactose permease, revealed by high sequence conservation (76%), overlapping substrate specificities, and similar transport kinetics, suggest a common origin of these transport systems. alpha-Galactosidase and sucrose hydrolase are not related to host enzymes but have their counterparts in other species. We propose a modular origin of the raf operon and discuss selective forces that favored the given gene organization also found in the E. coli lac operon. PMID- 2556374 TI - Molecular cloning of the 3' half of the Clostridium perfringens enterotoxin gene and demonstration that this region encodes receptor-binding activity. AB - Clostridium perfringens type A enterotoxin (CPE) causes the symptoms associated with C. perfringens food poisoning. To determine whether the C-terminal half of CPE contains receptor-binding activity, the 3' half of the cpe structural gene was cloned with an Escherichia coli expression vector system. E. coli lysates containing the expressed C-terminal CPE fragment (CPEfrag) were then assayed for CPE-like serologic, receptor-binding, and cytotoxic activities. CPEfrag was shown to contain an epitope located at or near the receptor-binding domain of the CPE molecule. Competitive-binding studies showed specific competition for CPE receptors between CPE and CPEfrag lysates. CPEfrag lysates did not cause cytotoxicity in Vero (African green monkey kidney) cells. However, preincubation with CPEfrag lysates specifically protected Vero cells from subsequent CPE challenge. This indicates that CPEfrag recognizes the physiologic receptor which mediates CPE cytotoxicity. Collectively, these studies indicate that the C terminal half of CPE contains a receptor-binding domain but additional amino acid sequences appear to be required for CPE cytotoxicity. PMID- 2556375 TI - Nucleotide sequence and insertional inactivation of a Bacillus subtilis gene that affects cell division, sporulation, and temperature sensitivity. AB - Located at 135 degrees on the Bacillus subtilis genetic map are several genes suspected to be involved in cell division and sporulation. Previously isolated mutations mapping at 135 degrees include the tms-12 mutation and mutations in the B. subtilis homologs of the Escherichia coli cell division genes ftsA and ftsZ. Previously, we cloned and sequenced the B. subtilis ftsA and ftsZ genes that are present on an 11-kilobase-pair EcoRI fragment and found that the gene products and organization of these two genes are conserved between the two bacterial species. We have since found that the mutation in the temperature-sensitive filamenting tms-12 mutant maps upstream of the ftsA gene on the same 11-kilobase pair EcoRI fragment in a gene we designated dds. Sequence analysis of the dds gene and four other open reading frames upstream of ftsA revealed no significant homology to other known genes. It was found that the dds gene is not absolutely essential for viability since the dds gene could be insertionally inactivated. The dds null mutants grew slowly, were filamentous, and exhibited a reduced level of sporulation. Additionally, these mutants were extremely temperature sensitive and were unable to form colonies at 37 degrees C. Another insertion, which resulted in the elimination of 103 C-terminal residues, resulted in a temperature sensitive phenotype less severe than that in the dds null mutant and similar to that in the known tms-12 mutant. The tms-12 mutation was cloned and sequenced, revealing a nonsense codon that was predicted to result in an amber fragment that was about 65% of the wild-type size (elimination of 93 C-terminal residues). PMID- 2556376 TI - Cloning and expression of a Bacillus subtilis division initiation gene for which a homolog has not been identified in another organism. AB - The Bacillus subtilis 168 division initiation genes defined by the temperature sensitive mutations ts-1 and ts-12 were cloned into a 10.5-kilobase EcoRI fragment of DNA in the lambda EMBL4 vector. The two genes were separated by approximately 3 kilobases. The gene in which the ts-1 mutation resides was shown to be the same as the B. subtilis homolog of the Escherichia coli ftsZ gene. The other gene was named divIB. It showed no homology to any previously identified gene and coded for a protein of 30.1 kilodaltons which was probably membrane bound. PMID- 2556377 TI - Suppression of the lethal effect of acidic-phospholipid deficiency by defective formation of the major outer membrane lipoprotein in Escherichia coli. AB - The Escherichia coli pgsA3 allele encoding a defective phosphatidylglycerophosphate synthase is lethal for all but certain strains. Genetic analysis of such strains has revealed that the lethal effect is fully suppressed by the lack of the major outer membrane lipoprotein that consumes phosphatidylglycerol for its maturation. PMID- 2556378 TI - Location of sites of transposon Tn916 insertion in the Mycoplasma mycoides genome. AB - The genome of Mycoplasma mycoides subsp. mycoides GC1176-2 was cleaved into six large fragments by the endonuclease KpnI which also cleaved the transposon Tn916 once. This has allowed genomic mapping of insertion sites for 50 transformants of GC1176-2 containing Tn916. Almost all of the mapped sites were clearly separate. The transformants provide a bank of genomes each with a KpnI site at a different position to facilitate mapping of gene loci. PMID- 2556379 TI - Functional domains of the gastric HK ATPase. AB - The gastric H+ + K+ ATPase is a member of the phosphorylating class of transport ATPase. Based on sequence homologies and CHO content, there may be a b subunit associated with the catalytic subunit of the H+ + K+ ATPase. Its function, if present, is unknown. The pump catalyzes a stoichiometric exchange of H+ for K+, but is also able to transport Na+ in the forward direction. This suggests that the transport step involves hydronium rather than protons. The initial binding site is likely to contain a histidine residue to account for the high affinity of the cellular site. The extracellular site probably lacks this histidine, so that a low affinity for hydronium allows release into a solution of pH 0.8. Labelling with positively charge, luminally reactive reagents that block ATPase and pump activity has shown that a region containing H5 and H6 and the intervening luminal loop is involved in necessary conformational changes for normal pump activity. The calculated structure of this loop shows the presence of an a helical, b turn, and b strand sector, with negative charges close to the membrane domain. This sector provides a possible site of interaction of drugs with the H+ + K+ ATPase, and may be part of the K+ pathway in the enzyme. PMID- 2556381 TI - SecB protein stabilizes a translocation-competent state of purified prePhoE protein. AB - Efficient translocation of pure precursor of PhoE protein (prePhoE) could be accomplished in an in vitro system consisting of only inverted Escherichia coli inner membrane vesicles, ATP, and SecA and SecB protein. In this in vitro system SecB and not trigger factor could stabilize a translocation-competent state of prePhoE. In contrast, translocation competency of proOmpA could be induced by both trigger factor and SecB protein, suggesting specificity in interactions between cytosolic factors and precursors in outer membrane protein translocation. PMID- 2556380 TI - Sodium, protons, and energy coupling in the methanogenic bacteria. AB - In this review, I focus on the bioenergetics of the methanogenic bacteria, with particular attention directed to the roles of transmembrane electrochemical gradients of sodium and proton. In addition, the mechanism of coupling ATP synthesis to methanogenic electron transfer is addressed. Evidence is reviewed which suggests that the methanogens possess great diversity in their bioenergetic machinery. In particular, in some methanogens the primary ion which is translocated coupled to metabolic energy is the proton, while others appear to utilize sodium. In addition, ATP synthesis driven by methanogenic electron transfer is accomplished in some organisms by a chemiosmotic mechanism and is coupled by a more direct mechanism in others. A possible explanation for this diversity (which is consistent with the relatedness of these organisms to each other and to other members of the Archaebacteria as determined by molecular biological techniques) is discussed. PMID- 2556382 TI - Dystrophin in electric organ of Torpedo californica homologous to that in human muscle. AB - We have found that dystrophin is highly concentrated at neuromuscular junctions and innervated membranes of the electric organ of Torpedo californica. In acetylcholine receptor-rich Torpedo membrane preparations dystrophin represents approximately 0.4% of total protein and can be extracted from these membranes by alkaline treatment in the absence of detergent, indicating that it is a peripheral membrane protein. Polyclonal antibodies raised against electrophoretically isolated Torpedo dystrophin cross-react with dystrophin in human muscle and unequivocally discriminate between normal and Duchenne muscular dystrophy patient's muscle. These results indicate that dystrophin is phylogenetically a highly conserved protein and that the relatively abundant dystrophin in electric organ would facilitate further investigations of its structure and function. PMID- 2556383 TI - Overexpression and assembly of the herpes simplex virus type 1 helicase-primase in insect cells. AB - Herpes simplex virus type 1 (HSV-1) encodes a helicase-primase that consists of three polypeptides encoded by the UL5, UL8, and UL52 genes (Crute, J.J., Tsurumi, T., Zhu, L., Weller, S.K., Olivo, P.D., Challberg, M.D., Mocarski, E.S., and Lehman, I.R. (1989) Proc. Natl. Acad, Sci, U.S.A. 86, 2186-2189). To obtain sufficient quantities of the enzyme for study, we have overexpressed the three genes using the baculovirus expression system. We find that the fully active enzyme can be assembled in vivo by triply infecting Spodoptera frugiperda SF9 cells with a baculovirus recombinant for each gene. The recombinant enzyme which we have purified to near homogeneity from the insect cells has a molecular weight of 270,000 and is composed of the three polypeptides encoded by the UL5, UL8, and UL52 genes. The enzyme possesses DNA-dependent ATPase, DNA-dependent GTPase, DNA helicase, and DNA primase activities that are essentially identical to the enzyme isolated from HSV-1-infected cells. PMID- 2556384 TI - Functional expression of the yeast alpha-factor receptor in Xenopus oocytes. AB - The STE2 gene of the yeast Saccharomyces cerevisiae encodes a 431-residue polypeptide that has been shown by chemical cross-linking and genetic studies to be a component of the receptor for the peptide mating pheromone, alpha-factor. To demonstrate directly that the ligand binding site of the alpha-factor receptor is comprised solely of the STE2 gene product, the STE2 protein was expressed in Xenopus oocytes. Oocytes microinjected with synthetic STE2 mRNA displayed specific surface binding for 35S-labeled alpha-factor (up to 40 sites/micron2/ng RNA). Oocytes injected with either STE2 antisense RNA or heterologous receptor mRNA (nicotinic acetylcholine receptor alpha, beta, gamma, and delta subunit mRNAs) showed no binding activity (indistinguishable from uninjected control oocytes). The apparent KD (7 nM) of the alpha-factor binding sites expressed on the oocyte surface, determined by competition binding studies, agreed with the values reported for intact yeast cells and yeast plasma membrane fractions. These findings demonstrate that the STE2 gene product is the only yeast polypeptide required for biogenesis of a functional alpha-factor receptor. Electrophysiological measurements indicated that the membrane conductance of oocytes injected with STE2 mRNA, or with both STE2 and GPA1 (encoding a yeast G protein alpha-subunit) mRNAs, did not change and was not affected by pheromone binding. Thus, the alpha-factor receptor, like mammalian G protein-coupled receptors, apparently lacks activity as an intrinsic or ligand-gated ion channel. This report is the first instance in which a membrane-bound receptor from a unicellular eukaryote has been expressed in a vertebrate cell. PMID- 2556385 TI - Activation of the serum response element and 12-O-tetradecanoylphorbol-13-acetate response element by the activated c-raf-1 protein in a manner independent of protein kinase C. AB - Transfection of the cDNA encoding the activated c-raf-1 protein or addition of 12 O-tetradecanoylphorbol-13-acetate (TPA) or dibutyryl cAMP to NIH/3T3 cells activated the c-fos gene enhancer linked to the chloramphenicol acetyltransferase or luciferase reporter gene. Prolonged treatment of NIH/3T3 cells with phorbol 12,13-dibutyrate caused down-regulation of protein kinase C. In these cells, addition of TPA did not stimulate the c-fos gene enhancer any more, but transfection of the c-raf-1 cDNA or addition of dibutyryl cAMP still stimulated the c-fos gene enhancer to the same extent as those induced in the control cells. Transfection of the c-raf-1 cDNA or addition of TPA to NIH/3T3 cells stimulated the serum response element and TPA response element but not the cAMP response element. In contrast, addition of dibutyryl cAMP to NIH/3T3 cells stimulated the cAMP response element but not the serum response element or TPA response element. These results indicate that the activated c-raf-1 protein stimulates the serum response element and TPA response element in a manner independent of protein kinase C and cAMP-dependent protein kinase. Since the c-fos gene enhancer has been shown to contain the serum response element and cAMP response element, it is most likely that the c-raf-1 protein is involved in the regulation of c-fos gene expression through the serum response element. PMID- 2556386 TI - Ca2(+)-mobilizing hormones stimulate Ca2+ efflux from hepatocytes. AB - Treatment of hepatocytes with 2,5-di-(tert-butyl)-1,4-benzohydroquinone (tBuBHQ), a novel mobilizer of the inositol 1,4,5-trisphosphate-sensitive Ca2+ pool, produces a sustained elevation of [Ca2+]i (Kass, G. E. N., Duddy, S. K., and Orrenius, S. (1989) J. Biol. Chem. 264, 15192-15198). Exposure of hepatocytes to the Ca2(+)-mobilizing hormones, vasopressin, angiotensin II, or ATP following [Ca2+]i elevation by tBuBHQ produced a rapid return of [Ca2+]i to basal or near basal levels. Release of the inositol 1,4,5-trisphosphate-sensitive Ca2+ pool by tBuBHQ following pretreatment with vasopressin or angiotensin II resulted in a [Ca2+]i transient and not the sustained [Ca2+]i elevation observed in the absence of the Ca2(+)-mobilizing hormones. The G-protein activator, NaF plus AlCl3, mimicked both effects of the Ca2(+)-mobilizing hormones on [Ca2+]i. The mechanism for Ca2+ removal from the cytosol by Ca2(+)-mobilizing hormones did not involve cyclic nucleotides nor did it require protein kinase C activation or cyclo- and lipoxygenase-dependent metabolites of arachidonic acid. Furthermore, the hormone mediated decrease in [Ca2+]i did not involve the pertussis toxin-sensitive Gi protein. Removal of the tBuBHQ-mobilized Ca2+ from the cytosol of hepatocytes by Ca2(+)-mobilizing hormones was mediated by stimulation of a Ca2+ efflux pathway. Thus, in addition to initiating [Ca2+]i transients by releasing Ca2+ from the inositol 1,4,5-trisphosphate-sensitive Ca2+ store and stimulating Ca2+ influx, Ca2(+)-mobilizing hormones also regulate the termination of the [Ca2+]i transient by stimulating a Ca2+ efflux pathway. PMID- 2556387 TI - G protein subunit, alpha i-3, activates a pertussis toxin-sensitive Na+ channel from the epithelial cell line, A6. AB - In nonpolar excitable cells, guanine nucleotide regulatory (G) proteins have been shown to modulate ion channel activity in response to hormone receptor activation. In polarized epithelia, hormone receptor-G protein coupling involved in the generation of cAMP occurs on the basolateral membrane, while the physiological response to this messenger is a stimulation of ion channel activity at the apical membrane. In the present study we have utilized the patch-clamp technique to assess if the polarized renal epithelia, A6, have topologically distinct G proteins at their apical membrane capable of modulating Na+ channel activity. In excised inside-out patches of apical membranes, spontaneous Na+ channel activity (conductance 8-9 picosiemens) was inhibited by the addition of 0.1 mM guanosine 5'-O-(2-thio)diphosphate to the cytosolic membrane surface without an effect on single channel conductance. In contrast, the percent open time of spontaneous Na+ channels increased from 6 to 50% following the addition of 0.1 mM GTP. The addition of preactivated pertussis toxin (100 ng/ml) to the cytosolic bathing solution of the excised patch inhibited spontaneous Na+ channel activity within a minute by 85% from approximately 47 to 7% open time and reduced the percent open time for Na+ channel activity to zero after approximately 3 min. The addition of 0.1 mM guanosine 5'-(3-O-thio)triphosphate or the addition of 20 pM purified human alpha i-3 subunit to pertussis toxin-treated membrane patches restored Na+ channel activity from zero to 35% open time. As little as 0.2 pM alpha i-3 subunit was capable of restoring Na+ channel activity. These data provide evidence for a role of pertussis toxin-sensitive G proteins in the apical plasma membrane of renal epithelia distal to signal transduction pathways in the basolateral membrane of these cells. This raises the possibility of a topologically distinct signal transducing pathway co-localized with the Na+ channel. PMID- 2556388 TI - Evidence for the direct interaction between tightly bound divalent metal ion and ATP on actin. Binding of the lambda isomers of beta gamma-bidentate CrATP to actin. AB - Competition between Ca2+ and Mg2+ for binding to a single high affinity site on actin has been confirmed. Occupancy of this site only by either Ca2+ or Mg2+ affects the conformation of actin and its ability to form nuclei and hydrolyze ATP. G-actin binds the beta gamma-bidentate CrATP, a substitution inert analog of metal-ATP complexes, and shows a high specificity for the lambda isomers. Binding of CrATP to ADP-actin is accompanied by the dissociation of tightly bound ADP and Ca2+. CrATP-actin shows a high tendency to form nuclei, like MgATP-actin. Polymerization of CrATP-actin is accompanied by cleavage of the gamma-phosphate, but subsequent Pi release cannot occur because the product of the reaction is the stable CrADP-Pi complex. All these results support the view that the divalent metal ion tightly bound to actin interacts with the beta- and gamma-phosphates of ATP in the nucleotide site. PMID- 2556389 TI - Lipid peroxyl radical intermediates in the peroxidation of polyunsaturated fatty acids by lipoxygenase. Direct electron spin resonance investigations. AB - Lipid peroxyl radicals resulting from the peroxidation of polyunsaturated fatty acids by soybean lipoxygenase were directly detected by the method of rapid mixing, continuous-flow electron spin resonance spectroscopy. When air-saturated borate buffer (pH 9.0) containing linoleic acid or arachidonate acid was mixed with lipoxygenase, fatty acid-derived peroxyl free radicals were readily detected; these radicals have a characteristic g-value of 2.014. An organic free radical (g = 2.004) was also detected; this may be the carbon-centered fatty acid free radical that is the precursor of the peroxyl free radical. The ESR spectrum of this species was not resolved, so the identification of this free radical was not possible. Fatty acids without at least two double bonds (e.g. stearic acid and oleic acid) did not give the corresponding peroxyl free radicals, suggesting that the formation of bisallylic carbon-centered radicals precedes peroxyl radical formation. The 3.8-G doublet feature of the fatty acid peroxyl spectrum was proven (by selective deuteration) to be a hyperfine coupling due to a gamma hydrogen that originated as a vinylic hydrogen of arachidonate. Arachidonate peroxyl radical formation was shown to be dependent on the substrate, active lipoxygenase, and molecular oxygen. Antioxidants are known to protect polyunsaturated fatty acids from peroxidation by scavenging peroxyl radicals and thus breaking the free radical chain reaction. Therefore, the peroxyl signal intensity from micellar arachidonate solutions was monitored as a function of the antioxidant concentration. The reaction of the peroxyl free radical with Trolox C was shown to be 10 times slower than that with vitamin E. The vitamin E and Trolox C phenoxyl radicals that resulted from scavenging the peroxyl radical were also detected. PMID- 2556390 TI - Magnetic circular dichroism and electron paramagnetic resonance studies of hydrogenases I and II from Clostridium pasteurianum. AB - The two iron-only hydrogenases (I and II) from Clostridium pasteurianum have been investigated by variable temperature magnetic circular dichroism (MCD) and electron paramagnetic resonance (EPR) spectroscopies. Samples were studied both reduced with dithionite under an atmosphere of H2 and after oxidation with thionine. The results are consistent with four and two [4Fe-4S]1+,2+ (F)-clusters in hydrogenases I and II, respectively. All four F-clusters are reduced and paramagnetic in reduced hydrogenase I, with up to one exhibiting an S = 3/2 ground state and the remainder having conventional S = 1/2 ground states. Both F clusters have S = 1/2 ground states in reduced hydrogenase II; however, one appears to be only partially reduced under the conditions used for reduction. MCD studies of the oxidized enzymes show no temperature-dependent features in the visible region which can be attributed to the EPR-active S = 1/2 hydrogen activating cluster, suggesting predominantly oxygen and nitrogen coordination for the iron atoms of this center. However, temperature-dependent MCD transitions arising from a hitherto undetected S greater than 1/2 Fe-S clusters are apparent in both oxidized hydrogenases. Detailed EPR studies of oxidized hydrogenase I revealed resonances from an S = 3/2 species, however, spin quantitation reveals this to be a trace component that is unlikely to be responsible for the observed low temperature MCD spectrum. The nature and origin of these S greater than 1/2 Fe-S clusters are discussed in light of the available spectroscopic data for these and other iron-only hydrogenases. PMID- 2556391 TI - Chimeric analogues of vertebrate gonadotropin-releasing hormones comprising substitutions of the variant amino acids in positions 5, 7, and 8. Characterization of requirements for receptor binding and gonadotropin release in mammalian and avian pituitary gonadotropes. AB - Variant forms of mammalian gonadotropin-releasing hormone (GnRH) (pGlu-His-Trp Ser-Tyr-Gly-Leu-Arg-Pro-Gly.NH2) are present in chicken ([Gln8] GnRH and [His5, Trp7, Tyr8]GnRH), salmon ([Trp7, Leu8]GnRH), and lamprey ([Tyr3, Leu5, Glu6, Trp7, Lys8] GnRH). To delineate the functional importance of the variant amino acids in positions 5, 7, and 8, the natural peptides and chimeric analogues were tested for gonadotropin-releasing activity and receptor-binding activity in rat, sheep, and chicken pituitaries. The results demonstrate that (i) the mammalian receptor has a high fidelity for Arg8 while the chicken receptor is less discriminatory and accepts basic or neutral amino acids in this position. Arg8 may contribute to conformational stabilization, and conformational constraint with D-Trp6 restored activity to analogues lacking Arg8 in the mammalian systems. D-Trp6 incorporation did not generally enhance activity in the chicken pituitary. (ii) His5 accompanying Arg8 in analogues markedly diminished activity in the chicken while gonadotropin-releasing activity was retained in the sheep pituitary. Receptor-binding activity was increased in the sheep indicating an uncoupling of receptor occupancy and activation. (iii) Substitution in position 7 is tolerated by the mammalian and chicken receptor. With Trp7-substituted analogues receptor-binding activity was relatively lower than gonadotropin releasing activity in the sheep pituitary, suggesting an enhanced receptor activation by these analogues or the existence of different GnRH receptors. PMID- 2556392 TI - Isolation and characterization of a putative collagen receptor from Staphylococcus aureus strain Cowan 1. AB - In a previous study we demonstrated that cells of Staphylococcus aureus strain Cowan bind 125I-collagen in a receptor-ligand type of interaction (Speziale, P., Raucci, G., Visai, L., Switalski, L.M., Timpl, R., and Hook, M. (1986) J. Bacteriol. 167, 77-81). In the present communication we report on the isolation and preliminary characterization of a putative collagen receptor from a lysate of S. aureus strain Cowan. Antibodies raised against a collagen receptor positive strain inhibit the binding of 125I-collagen to bacterial cells, whereas antibodies raised against a collagen receptor negative strain were without effect. Solubilized cell surface components did not exhibit any measurable affinity for collagen-Sepharose. However, the inhibitory effect of the antibodies against bacterial cells was neutralized by the lysate from a receptor-positive but not receptor-negative strain. A collagen receptor assay was designed based on this observation and used to develop a receptor purification protocol involving anion exchange chromatography, ammonium sulfate precipitation, and gel chromatography. Using this procedure a protein with an apparent Mr of 135,000 was purified. This protein which was present on a collagen receptor-positive strain but not on a receptor-negative strain could completely neutralize the inhibitory activity of the antibodies raised against S. aureus strain Cowan. Furthermore, antibodies raised against the 135-kDa protein inhibited the binding of collagen to bacteria, and this protein is tentatively identified as a collagen receptor. PMID- 2556393 TI - Increase in Gs and cyclic AMP generation in HIT cells. Evidence that the 45-kDa alpha-subunit of Gs has greater functional activity than the 52-kDa alpha subunit. AB - Cyclic AMP accumulation in response to forskolin, cholera toxin, or isoproterenol is dramatically increased in HIT T-15 cells, a clonal cell line of Syrian hamster pancreatic islet beta cells, as a function of passage number. Forskolin and cholera toxin elevate cyclic AMP levels 5- to 10-fold higher in later passages (87-100) than in earlier passages (70-80). A similar phenomenon is observed with isoproterenol (10 microM) which increases cyclic AMP levels 56-fold in older HIT cells (passage 94), whereas only marginally stimulating cyclic AMP production in younger cells (passage 70-82). To determine whether a change in the stimulatory or inhibitory guanine nucleotide regulatory proteins, Gs or Gi, was responsible for these observations, ADP-ribosylation of HIT cell membranes with cholera toxin and pertussis toxin was examined. All passages contained two cholera toxin substrates at 52 and 45 kDa. The amount of 52 kDa did not appear to change with passage number, but the amount of 45 kDa increased in the later passages (89 and 94). The ratio of 45 to 52 kDa cholera toxin substrate, as determined by densitometric analysis, increased from 0.1 in passages 70, 75, and 82 to 0.45 at passage 89. No passage related changes in a 40-kDa pertussis toxin substrate were observed. An increase in the amount of the 45-kDa alpha-subunit of Gs was confirmed on immunoblots using antisera specific for the alpha-subunits of Gs. The amount of functional Gs present in various HIT cell passages was examined by determining the extent to which extracts from HIT cell membranes reconstituted guanine nucleotide-sensitive adenylyl cyclase in S49 cyc- membranes. Extracts derived from passage 94 reconstituted three to four times more adenylyl cyclase activity in cyc- membranes than extracts from passages 70, 75, and 82. These data indicate that an increase in functional Gs in later passages may be the underlying cause for the increased responsiveness to isoproterenol and forskolin in later passages. These data also suggest that functional differences exist between the Gs alpha-subunits, with the smaller 45-kDa subunit being more efficacious in coupling to cyclic AMP synthesis than the larger 52-kDa subunit. This is a departure from the commonly held view that the two subunits have similar efficacies in stimulating adenylyl cyclase. PMID- 2556394 TI - Valyl-tRNA synthetase from rabbit liver. I. Purification as a heterotypic complex in association with elongation factor 1. AB - Valyl-tRNA synthetase occurs as a high molecular mass entity of approximately equal to 700 kDa in the crude extract from rabbit liver. The enzyme was purified as a heterotypic complex comprising four polypeptides of 140, 50, 35, and 27 kDa in the molar proportions of 1:2:1:1, respectively, as determined by one dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Co purification of these components at each step of the purification supports the conclusion that they are physically associated within the same complex. In addition to valyl-tRNA synthetase activity, which was assigned to the 140-kDa component, the purified complex exhibits a potent Elongation Factor 1 activity, determined by its ability to sustain poly(U)-dependent polyphenylalanine synthesis in the presence of Elongation Factor 2. Our results are essentially in agreement with those from a recent report (Motorin, Y., Wolfson, A., Orlovsky, A., and Gladilin, K. (1988) FEBS Lett. 238, 262-264), according to which the polypeptides other than that assigned to valyl-tRNA synthetase correspond to the subunits of Elongation Factor 1H. PMID- 2556395 TI - Valyl-tRNA synthetase from rabbit liver. II. The enzyme derived from the high-Mr complex displays hydrophobic as well as polyanion-binding properties. AB - The preceding paper (Bec, G., Kerjan, P., Zha, X.D., and Waller, J.P. (1989) J. Biol. Chem. 264, 21131-21137) described the purification to apparent homogeneity from rabbit liver, of a heterotypic complex comprising valyl-tRNA synthetase and Elongation Factor 1H. In the present study, valyl-tRNA synthetase was dissociated and separated from the other components of this complex by hydroxylapatite chromatography in the presence of 0.5 M NaSCN. The properties of the homogeneous mammalian enzyme were compared to those of the corresponding enzyme from yeast. Both behaved as monomeric entities, with apparent molecular masses of 140 and 125 kDa, respectively. Furthermore, both displayed strong affinity toward the polyanionic support heparin-Ultrogel, a property not manifested by the corresponding prokaryotic enzyme. However, unlike the yeast enzyme, that of mammalian origin additionally exhibited hydrophobic properties, as reflected by its affinity toward phenyl-Sepharose. A structural model is proposed according to which mammalian valyl-tRNA synthetase has conserved the polycationic N-terminal domain that distinguishes the corresponding lower eukaryotic enzyme from its prokaryotic counterpart, while acquiring a hydrophobic domain most likely responsible for its association to Elongation Factor 1H. PMID- 2556396 TI - Characterization of the relA1 mutation and a comparison of relA1 with new relA null alleles in Escherichia coli. AB - The most widely studied "relaxed" mutant of the relA locus, the relA1 allele, is shown here to consist of an IS2 insertion between the 85th and 86th codons of the otherwise wild-type relA structural gene, which normally encodes a 743-amino acid (84 kDa) protein. The RelA protein is a ribosome-dependent ATP:GTP (GDP) pyrophosphoryltransferase that is activated during the stringent response to amino acid starvation and thereby occasions the accumulation of guanosine 3',5' bispyrophosphate (ppGpp). We propose that the IS2 insertion functionally splits the RelA protein into two (alpha and beta) peptide fragments which can complement each other in trans to yield residual ppGpp synthetic activity; neither fragment shows this activity when expressed alone. Cell strains with a single copy relA null allele show physiological behavior that is much the same as relA1 mutant strains. Both relA1 and relA null strains accumulate ppGpp during glucose starvation and do not accumulate ppGpp during the stringent response. The presence of ppGpp in verifiable relA null strains is interpreted as unequivocal evidence for an alternate route of ppGpp synthesis that exists in addition to the relA-dependent reaction. PMID- 2556397 TI - Defective homologous pairing and proficient processive unwinding by the recA430 mutant protein and intermediates of homologous pairing by recA protein. AB - The recA protein promotes the formation and processing of joint molecules of homologous double- and single-stranded DNAs in vitro. Under a set of specified conditions, we found that the substitution of a single amino acid in the recA protein (recA430 mutation) depresses its activity for the homologous pairing to about 1/100 of that by the wild type protein when compared by the rate for the first 2-3 min of the reaction, but that the mutation only slightly, if at all, affects its ability to bind progressively to double-stranded DNA to unwind the double helix ("processive unwinding"). This is in striking contrast to an anti recA protein monoclonal IgG, ARM193, which severely inhibits the processive unwinding but not the homologous pairing, providing further support for our conclusion that the homologous pairing and processive unwinding are functionally independent of each other. Antibody ARM193 caused the breakdown of spontaneously formed filaments of the recA protein, but the recA430 mutation did not affect the self-polymerization of the protein. The recA430 protein was apparently proficient in the functional binding to a single-stranded DNA and in the hydrolysis of ATP. However, we found that under the above conditions the mutant protein was defective as to homology-independent conjunction of DNA molecules to form a "ternary complex" (of macromolecules). These results suggest that (i) only one DNA-binding site is sufficient for the recA protein to promote the processive unwinding (the ability of the protein to form spontaneous filaments is closely related to this process) and that (ii) two DNA-binding sites on each of the recA polypeptides or those composed of a dimer (or oligomer) of the polypeptide are required for the recA protein to promote both the conjunction of parental DNA molecules and the homologous pairing (the ability to form the spontaneous filaments is not essential to this process). (iii) The simultaneous inactivation of the activity to promote the homologous pairing and that to form the ternary complex by the single substitution of the amino acid provides a physical support for the conclusion that the ternary complex is an indispensable intermediate in the homologous pairing. PMID- 2556398 TI - Apolipoprotein E3-Leiden contains a seven-amino acid insertion that is a tandem repeat of residues 121-127. AB - Apolipoprotein (apo) E3-Leiden is a variant of apoE that is associated with dominant expression of type III hyperlipoproteinemia and that is defective in binding to the low density lipoprotein receptor. Therefore, the structure of apoE3-Leiden was investigated. Upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis apoE3-Leiden and its 22-kDa amino-terminal thrombolytic fragment migrated with a higher than normal apparent molecular weight. The structural abnormality of apoE3-Leiden was determined by sequencing its CNBr-, tryptic-, and Staphylococcus aureus V8 protease-generated peptides. In contrast to normal apoE3, which has a cysteine at residue 112, apoE3-Leiden does not contain any cysteine and has an arginine at position 112 (as does apoE4, which also completely lacks cysteine). The basis for the molecular weight difference was determined to be a seven-amino acid insertion that is a tandem repeat of residues 121-127 of normal apoE3, i.e. Glu-Val-Gln-Ala-Met-Leu-Gly, resulting in apoE3 Leiden having 306 amino acids rather than 299. The negatively charged glutamyl residues within the insertion compensates for the arginine substitution at residue 112; thus apoE3-Leiden focuses in the E3 position. The low density lipoprotein receptor binding activities of both intact apoE3-Leiden and its 22 kDa thrombolytic fragment were determined in an in vitro assay. Although apoE3 Leiden had only about 25% of normal binding activity, its 22-kDa thrombolytic fragment had nearly normal binding, suggesting that the carboxyl-terminal domain of apoE3-Leiden modulates the receptor binding function of its amino-terminal domain. PMID- 2556399 TI - Photoaffinity labeling of the purified skeletal muscle calcium antagonist receptor by a novel benzothiazepine, [3H]azidobutyryl diltiazem. AB - Previous photoaffinity-labeling studies with [3H]azidopine, (+) [3H]PN200-110, and [3H]LU 49888 have demonstrated that 1,4-dihydropyridines (nifedipine-like drugs) and phenylalkylamines (verapamil-like drugs) bind exclusively to the 165 kDa alpha 1 subunit of skeletal muscle calcium channels. However, it has not been conclusively determined whether benzothiazepines (diltiazem-like drugs), which represent the third group of calcium antagonists, also bind to the alpha 1 subunit. Here we report data obtained with a newly developed benzothiazepine photoaffinity probe, [3H]azidobutyryl diltiazem. This drug competes with diltiazem for the benzothiazepine-binding site and, in purified calcium channel preparations, specifically labels the 165-kDa polypeptide which does not change its electrophoretic mobility upon disulfide reduction. These data show that benzothiazepines, just like 1,4-dihydropyridines and phenylalkylamines, bind to the alpha 1 subunit of the skeletal muscle calcium channels. PMID- 2556400 TI - Phosphorylation of cellular proteins regulates their binding to the cAMP response element. AB - We have studied the protein factors that promote transcription via binding to the cAMP response element (CRE) present in the adenovirus early region III (EIII) and early region IV (EIV) promoters. Three sets of CRE-binding phosphoproteins, ranging in molecular mass from 65-72, 38-43, and 31-37 kDa, were identified in vivo from HeLa cells. Western blot analysis revealed that all three sets of proteins identified were immunologically related to the transcription factor AP1. We found that binding of these proteins to the CRE could be regulated by phosphorylation in vitro. EivF, a 65-72-kDa protein was found to bind specifically to the adenovirus EIV promoter. We have also shown that the smaller molecular mass proteins of 31-37 and 38-43 kDa were able to bind to the CRE present in the adenovirus EIV promoter, as well as to two related DNA elements present in the adenovirus EIII promoter, the ATF and AP1 sites. Phosphorylation of these proteins with the cAMP-dependent protein kinase, affected their transcriptional activity and binding affinity to the three sites. Furthermore, the binding specificity of the 31-37-kDa polypeptides was mediated by cAMP dependent protein kinase in vitro. Our data suggests that phosphorylation of factors that bind to the CRE may, in part, underlie the cellular response to the adenovirus-encoded Ela protein. PMID- 2556401 TI - Purification and characterization of a DNA-pairing and strand transfer activity from mitotic Saccharomyces cerevisiae. AB - An enzyme catalyzing homologous pairing of DNA chains has been extensively purified from mitotic yeast. The most highly purified fractions are enriched for a polypeptide with a molecular mass of approximately 120 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Protein-dependent pairing of single-stranded DNAs requires a divalent cation (Mg2+ or Ca2+) but proceeds rapidly in the absence of any nucleoside triphosphates. The kinetics of reassociation are extremely rapid, with more than 60% of the single-stranded DNA becoming resistant to S1 nuclease within 1 min at a ratio of 1 protein monomer/50 nucleotides. The results of enzyme titration and DNA challenge experiments suggest that this protein does not act catalytically during renaturation but is required stoichiometrically. The protein promotes formation of joint molecules between linear M13 replicative form DNA (form III) containing short single stranded tails and homologous single-stranded M13 viral DNA. Removal of approximately 50 nucleotides from the ends of the linear duplex using either exonuclease III (5' ends) or T7 gene 6 exonuclease (3' ends) activates the duplex for extensive strand exchange. Electron microscopic analysis of product molecules suggests that the homologous circular DNA initially associates with the single stranded tails of the duplexes, and the heteroduplex region is extended with displacement of the noncomplementary strand. The ability of this protein to pair and to promote strand transfer using either exonuclease III or T7 gene 6 exonuclease-treated duplex substrates suggests that this activity promotes heteroduplex extension in a nonpolar fashion. The biochemical properties of the transferase are consistent with a role for this protein in heteroduplex joint formation during mitotic recombination in Saccharomyces cerevisiae. PMID- 2556402 TI - Protein import into mitochondria: ATP-dependent protein translocation activity in a submitochondrial fraction enriched in membrane contact sites and specific proteins. AB - To identify the membrane regions through which yeast mitochondria import proteins from the cytoplasm, we have tagged these regions with two different partly translocated precursor proteins. One of these was bound to the mitochondrial surface of ATP-depleted mitochondria and could subsequently be chased into mitochondria upon addition of ATP. The other intermediate was irreversibly stuck across both mitochondrial membranes at protein import sites. Upon subfraction of the mitochondria, both intermediates cofractionated with membrane vesicles whose buoyant density was between that of inner and outer membranes. When these vesicles were prepared from mitochondria containing the chaseable intermediate, they internalized it upon addition of ATP. A non-hydrolyzable ATP analogue was inactive. This vesicle fraction contained closed, right-side-out inner membrane vesicles attached to leaky outer membrane vesicles. The vesicles contained the mitochondrial binding sites for cytoplasmic ribosomes and contained several mitochondrial proteins that were enriched relative to markers of inner or outer membranes. By immunoelectron microscopy, two of these proteins were concentrated at sites where mitochondrial inner and outer membranes are closely apposed. We conclude that these vesicles contain contact sites between the two mitochondrial membranes, that these sites are the entry point for proteins into mitochondria, and that the isolated vesicles are still translocation competent. PMID- 2556403 TI - Signal recognition particle mediates a transient elongation arrest of preprolactin in reticulocyte lysate. AB - Signal recognition particle (SRP) is a ribonucleoprotein that functions in the targeting of ribosomes synthesizing presecretory proteins to the ER. SRP binds to the signal sequence as it emerges from the ribosome, and in wheat germ extracts, arrests further elongation. The translation arrest is released when SRP interacts with its receptor on the ER membrane. We show that the delay of elongation mediated by SRP is not unique to wheat germ translation extracts. Addition of mammalian SRP to reticulocyte lysates resulted in a delay of preprolactin synthesis due to increased ribosome pausing at specific sites on preprolactin mRNA. Addition of canine pancreatic microsomal membranes to reticulocyte lysates resulted in an acceleration of preprolactin synthesis, suggesting that the endogenous SRP present in the reticulocyte lysate also delays synthesis of secretory proteins. PMID- 2556404 TI - A yeast gene important for protein assembly into the endoplasmic reticulum and the nucleus has homology to DnaJ, an Escherichia coli heat shock protein. AB - When nuclear localization sequences (termed NLS) are placed at the N terminus of cytochrome c1, a mitochondrial inner membrane protein, the resulting hybrid proteins do not assemble into mitochondria when synthesized in the yeast Saccharomyces cerevisiae. Cells lacking mitochondrial cytochrome c1, but expressing the hybrid NLS-cytochrome c1 proteins, are unable to grow on glycerol since the hybrid proteins are associated primarily with the nucleus. A similar hybrid protein with a mutant NLS is transported to and assembled into the mitochondria. To identify proteins that might be involved in recognition of nuclear localization signals, we isolated conditional-lethal mutants (npl, for nuclear protein localization) that missorted NLS-cytochrome c1 to the mitochondria, allowing growth on glycerol. The gene corresponding to one complementation group (NPL1) encodes a protein with homology to DnaJ, an Escherichia coli heat shock protein. npl1-1 is allelic to sec63, a gene that affects transit of nascent secretory proteins across the endoplasmic reticulum. Rothblatt, J. A., R. J. Deshaies, S. L. Sanders, G. Daum, and R. Schekman. 1989. J. Cell Biol. 109:2641-2652. The npl1 mutants reported here also weakly affect translocation of preprocarboxypeptidaseY across the ER membrane. A normally nuclear hybrid protein containing a NLS fused to invertase and a nucleolar protein are not localized to the nucleus in npl1/sec63 cells at the nonpermissive temperature. Thus, NPL1/SEC63 may act at a very early common step in localization of proteins to the nucleus and the ER. Alternatively, by affecting ER and nuclear envelope assembly, npl1 may indirectly alter assembly of proteins into the nucleus. PMID- 2556405 TI - Endocytosis of simian virus 40 into the endoplasmic reticulum. AB - The endocytosis of SV-40 into CV-1 cells we studied using biochemical and ultrastructural techniques. The half-time of binding of [35S]methionine radiolabeled SV-40 to CV-1 cells was 25 min. Most of the incoming virus particles remained undegraded for several hours. Electron microscopy showed that some virus entered the endosomal/lysosomal pathway via coated vesicles, while the majority were endocytosed via small uncoated vesicles. After infection at high multiplicity, one third of total cell-associated virus was observed to enter the ER, starting 1-2 h after virus application. The viruses were present in large, tubular, smooth membrane networks generated as extentions of the ER. The results describe a novel and unique membrane transport pathway that allows endocytosed viral particles to be targeted from the plasma membrane to the ER. PMID- 2556406 TI - Distinct endocytotic pathways in epidermal growth factor-stimulated human carcinoma A431 cells. AB - Addition of EGF to human epidermoid carcinoma A431 cells increases the rate of fluid-phase pinocytosis 6-10-fold as measured by horseradish peroxidase uptake (Haigler, H.T., J. A. McKanna, and S. Cohen. 1979. J. Cell Biol. 83:82-90). We show here that in the absence of extracellular Na+ or in the presence of amiloride the stimulation of pinocytosis by EGF is substantially reduced. Amiloride had no effect on the endocytosis of EGF itself or of transferrin, demonstrating that the receptor-mediated endocytotic pathway operated normally under conditions that blocked stimulated pinocytosis. Amiloride blocked EGF stimulated pinocytosis in both HCO3(-)-containing and HCO3(-)-free media. The EGF stimulated pinocytotic activity can frequently be localized to areas of the cell where membrane spreading and ruffling are taking place. These results demonstrate that (a) EGF induces a distinct amiloride-sensitive endocytotic pathway on A431 cells; (b) occupied EGF receptors do not utilize this pathway for their own entry; (c) endocytosis of occupied EGF receptors is not in itself sufficient to stimulate pinocytosis. PMID- 2556407 TI - cAMP-mediated inhibition of intracellular particle movement and actin reorganization in Dictyostelium. AB - Before addition of cAMP, Dictyostelum amoebae rapidly translocating in buffer are elongate, exhibit expansion zones primarily at the anterior end and filamentous actin (F-actin) localization primarily in the anterior pseudopodia. Intracellular particle movement is primarily in the anterior direction, and the average rate of particle movement is roughly five times the rate of cellular translocation. Within seconds after the addition of 10(-6)M cAMP, there is a dramatic suppression of cellular translocation, an inhibition of pseudopod formation, a freeze in cellular morphology, a dramatic depression in intracellular particle movement, loss of F-actin localization in pseudopodia concomitant with relocalization of F-actin in the general cytoplasmic cortex under the plasma membrane, and a doubling of F-actin content. After 10 s, expansion zones are again visible at the cell perimeter, but they no longer are localized in the original anterior portion of the cell. There is a slight rebound in particle movement after 10 s, but particles with persistent tracks now show no directionality towards the original anterior portion of the cell, as they did before cAMP addition. Finally, in parallel with the resumption of peripheral expansion and the small rebound in particle movement, there is a decrease in total cellular F-actin to the untreated level. The pattern of microtubule organization is unaffected by the addition of cAMP. PMID- 2556409 TI - Cytochemical and molecular properties of simian virus 40 transformed Kaposi's sarcoma-derived cells: evidence for the secretion of a member of the fibroblast growth factor family. AB - Contact-inhibited Kaposi's sarcoma-derived cells (KS cells) were transfected with Simian Virus 40 (SV40) DNA. Transformed cells (SV-KSC) were selected for their capacity to form foci on monolayers of the low-malignant KS cells. Isolated SV KSC foci were found to contain integrated SV40 DNA sequences and to express SV40 large T-antigen. Several differentiation properties of KS cells are retained in the SV40 transformants, e.g., expression of vimentin and the endothelial cell marker BMA 120. In contrast to the maternal KS cells, SV-KSC are capable of growing in platelet-derived growth factor (PDGF)-depleted platelet-poor-plasma serum (PPPS) and in soft agar. However, they are not tumorigenic in nude mice. Expression of the oncogenes c-myc, c-N-ras, c-Ha-ras, and p53 is significantly elevated in SV-KSC, whereas c-fos and c-erb B expression is comparable to that of KS cells and fibroblasts. Conditioned medium from SV-KSC can substitute for PDGF when PDGF-dependent, nontransformed KS cells are grown in PPPS. Biochemical analysis of the SV-KSC supernatant and PDGF A and B mRNA expression analysis provide evidence that the mitogenic activity is not due to a PDGF-like growth factor. On the other hand, there is evidence to indicate that the SV-KSC mitogen is a member of the fibroblast growth factor family. SV-KSC represent an interesting model system for the study of different degrees of malignancy of cultured mesenchymal cells and especially provide an important source for the isolation of a potent growth factor for KS cells and other mesenchymal cells in vitro. PMID- 2556410 TI - Basic fibroblast growth factor: production, mitogenic response, and post-receptor signal transduction in cultured normal and transformed fetal bovine aortic endothelial cells. AB - Normal FBAE AG 7680 cells and chemically transformed FBAE GM 7373 cells were compared for their capacity to produce and to respond to bFGF. Normal FBAE cells showed higher levels of bFGF protein and of poly(A)+ bFGF mRNA than transformed GM 7373 cells, indicating that chemical transformation in FBAE cells is paralleled by a decrease of bFGF gene expression. Basic FGF induced cell proliferation in both normal and transformed FBAE cells. However, bFGF appeared to be much more potent in transformed than in normal cells. No differences in bFGF membrane receptors were observed between normal and transformed FBAE cells in terms of apparent molecular weight, number per cell, dissociation constant, and kinetic of downregulation. In respect to normal cells, however, transformed GM 7373 cells showed higher basal levels of PKC activity. This kinase is activated by bFGF and is involved in mediating the mitogenic activity of bFGF, as shown by the capacity of the PKC inhibitor H-7 to abolish the mitogenic activity of bFGF both in normal and transformed FBAE cells. Like bFGF, the PKC activators DAG and TPA exerted a stronger mitogenic activity in transformed than in normal FBAE cells. Thus, the different susceptibility of normal and transformed FBAE cells to bFGF appears to depend on differences in the post-receptor signal transduction mediated by PKC rather than on differences in bFGF receptors. The results indicate that chemical transformation causes significant modifications of bFGF physiology in FBAE cells. The relevance of these modifications to the genesis of tumors of vascular origin deserves further investigation. PMID- 2556408 TI - Transformation of murine melanocytes by basic fibroblast growth factor cDNA and oncogenes and selective suppression of the transformed phenotype in a reconstituted cutaneous environment. AB - Constitutive expression of basic fibroblast growth factor (bFGF), a common characteristic of metastatic melanomas, was reproduced in vitro by infection of normal murine melanocytes with a recombinant retrovirus carrying a cDNA for bFGF. Expression of bFGF in these cells conferred autonomous growth in culture and extinguished differentiated functions, such as the synthesis of melanin and formation of dendrites. Independence from exogenous bFGF and loss of differentiated functions in vitro were induced also by transformation of melanocytes with the oncogenes myc, Ela, ras, and neu, although bFGF was not expressed by the respective transformants. As shown in skin reconstitution experiments onto syngeneic mice and subcutaneous injections into nude mice, the various transformants differed in their behavior in vivo. The bFGF transformants did not form tumors. They reverted to having a normal, melanotic phenotype and restricted growth. Myc and Ela transformants grew as tumors in nude mice but not in syngeneic, immunocompetent animals. Ras-transformed melanocytes were always tumorigenic, whereas the formation of tumors by neu transformants was suppressed by the concomitant grafting of keratinocytes in reconstituted skin of syngeneic mice. These data show that melanocytes genetically manipulated to produce bFGF acquire properties in vitro similar to those of metastatic melanoma cells or those induced by various oncogenes but that constitutive production of bFGF by itself is insufficient to make melanocytes tumorigenic. The experiments also show that melanocytes transformed by the selected oncogenes respond differentially to various environments in vivo. PMID- 2556412 TI - Pulmonary microvascular endothelial cell contractility on silicone rubber substrate. AB - Endothelial cell (EC) motility may contribute to the regulation of microvascular perfusion and/or paracellular permeability. The experiments reported herein demonstrate that bovine pulmonary microvessel EC can reversibly deform a silicone substrate in response to agents known to contract and relax smooth muscle cells. Contracting pulmonary microvessel EC exerted a tension that created wrinkles in the underlying deformable substrate. Relaxation and loss of tension were revealed by the disappearance of these wrinkles without loss of cell adhesion to the substratum. Angiotensin II (Ang II) and bradykinin stimulated pulmonary microvessel EC to contract within 3 to 8 min in a Ca2+-dependent fashion. The peak of contraction at 10 to 20 min was followed by relaxation. Forskolin and sodium nitroprusside (SNP) initiated relaxation of the microvessel EC within 3 to 10 min respectively. Relaxed EC contracted following the addition of Ang II, also within 3 min. Dibutyryl cAMP, dibutyryl cGMP, and the photoactivated internalized "caged" cAMP and cGMP promoted EC relaxation in a manner similar to forskolin and SNP. Increases in the intracellular concentration of inositol triphosphate (IP3) with the photoactivated IP3 complex promoted EC contraction in 2 min with a peak at 7 min. The contraction was followed by relaxation, which occurred at 20-25 min. Neither bovine pulmonary artery nor retinal microvessel EC, used as controls, contracted under these experimental conditions. One could speculate that this unique contractile property of pulmonary microvessel EC as observed in vitro may play a regulatory role in vivo, in local perfusion and/or in intercellular gap regulation. PMID- 2556411 TI - Activation and proliferation signals in murine macrophages: relationships among c fos and c-myc expression, phosphoinositide hydrolysis, superoxide formation, and DNA synthesis. AB - Murine bone marrow-derived macrophages (BMM) undergo DNA synthesis in response to growth factors such as colony stimulating factor-1 (CSF-1) and granulocyte macrophage CSF (GM-CSF). These macrophages can also be "activated," but without subsequent DNA synthesis, by a number of other agents, including lipopolysaccharide (LPS), concanavalin A, zymosan, formyl-methionyl-leucyl phenylalanine (FMLP), and the Ca2+ ionophore, A23187. When BMM are treated with a range of stimuli, there is some, although not perfect, correlation between transient elevations in both c-myc mRNA and c-fos mRNA levels and increases in DNA synthesis. However, enhanced DNA synthesis and oncogene expression are readily dissociated from rises in inositol phosphates and, by implication, phospholipase C-mediated hydrolysis of phosphatidyl inositol 4,5-bisphosphate. Superoxide formation in BMM can also be dissociated from the other responses and does not necessarily depend on protein kinase C activation. PMID- 2556413 TI - Collagenase secretion accompanying changes in cell shape occurs only in the presence of a biologically active cytokine. AB - We have investigated the relationship between collagenase production, cell shape and stimulatory factors in cell culture. In a homogeneous culture of primary rabbit corneal stromal cells, shape change induced by a variety of agents was not effective in stimulating collagenase secretion. Only in the presence of a biologically active cytokine or phorbol myristate acetate was a correlation seen between changes in cell shape (induced by a second agent) and collagenase secretion by these primary cells. Cell shape changes were not, however, necessary for collagenase secretion, since certain concentrations of endotoxin or lactalbumin hydrolysate effected secretion of the enzyme in the absence of morphological changes. With passaged cells or mixed cell cultures, where cell shape change did correlate with collagenase secretion without the addition of an exogenous agent, the production of an effective cytokine (autocrine or paracrine) was demonstrated. Thus cell shape change seems to be neither universally necessary nor sufficient for the stimulation of collagenase secretion. It is proposed that the function of cytokines may be more immediately related to gene expression in this system than is change in the shape of the cell. The hypothesis is presented that cell shape changes may render the target cells receptive to cytokines, perhaps by replacing the need for a natural cytokine cofactor. It is also demonstrated here that the use of passaged cells, mixed cell cultures containing endogenous cytokine-secreting cells or tissue culture additives can profoundly affect the interpretation of the effect of various agents on collagenase secretion, and may lead to observations that are not directly relevant to cell function in vivo. PMID- 2556414 TI - Gelatinase (type IV collagenase) immunolocalization in cells and tissues: use of an antiserum to rabbit bone gelatinase that identifies high and low Mr forms. AB - An antiserum was raised to rabbit bone gelatinase (type IV collagenase). It was shown by immunoblotting to detect both the low Mr (72,000) enzyme produced by connective tissue cells from rabbit, pig, human and mouse, as well as the high Mr (94,000-97,000) enzymes secreted by macrophages and polymorphonuclear leucocytes from these species, and by rabbit chondrocytes and endothelial cells. Crossed immunoblotting, antibody inhibition and deglycosylation studies indicated that the high and low Mr forms of gelatinase are immunologically distinct gene products, although their substrate specificity profiles are identical. The anti gelatinase antiserum was used to immunolocalize the enzyme. Gelatinase was most efficiently detected in rabbit monocytes and connective tissue cells, but cells derived from the human and pig gave poor immunostaining, although mouse gelatinase stained well. The anti-gelatinase antiserum stained cells of the synovial tissue of rabbits at 14 days after induction of an antigen-induced arthritis, demonstrating its usefulness as a tool to assess the role of this enzyme in degradative events. PMID- 2556415 TI - [Krukenberg's tumor: a still unfavorable prognosis]. PMID- 2556416 TI - Performance of carbohydrate-modified fused-silica capillaries for the separation of proteins by zone electrophoresis. AB - The walls of fused-silica capillaries were chemically modified with small carbohydrate moieties in order to diminish the wall adsorption of proteins in capillary zone electrophoresis. A diol-type coating, prepared by bonding of gamma glycidoxy propyltrimethoxysilane to the wall followed by acidic hydrolysis, shows for proteins a similar electrophoretic behaviour as various pH values to a polyethylene glycol (PEG) coating tested previously. Although good peak shapes were obtained for proteins in the pH range 3-5, the efficiency on the diol coating is worse than that on the PEG coating. At higher pH values the peaks are deformed and the efficiency is lost. A maltose coating appears to shield the silica surface well for proteins up to pH 7. The peak shapes of proteins are acceptable, but the efficiency of the maltose coating is smaller than that on the diol coating. The diol coating is stable in the indicated pH range. However, with the maltose coating good stability is obtained only on adding an antimicrobial agent to the buffers. PMID- 2556417 TI - Development and validation of a liquid chromatographic-mass spectrometric assay for the determination of sumatriptan in plasma. AB - Sumatriptan succinate is a novel compound currently in development for the acute treatment of migraine. During early studies in man a sensitive and selective assay was required, which had to be developed rapidly, to determine plasma concentrations following an intravenous infusion. Thermospray liquid chromatography-mass spectrometry combined with the advanced automated sample processor was selected to achieve this. Although the assay was required quickly criteria for intra- and inter-assay accuracies and precisions of +/- 10% had to be achieved. These were obtained only by using a co-eluting deuterium-labelled internal standard. Attempts to use a homologue as an internal standard, which did not co-elute with sumatriptan, gave inferior results. The assay was linear over the calibration range 2-50 ng/ml with a limit of quantification of 2 ng/ml. The application of the technique to the analysis of samples from a volunteer study is demonstrated. PMID- 2556418 TI - Determination of tazobactam and piperacillin in human plasma, serum, bile and urine by gradient elution reversed-phase high-performance liquid chromatography. AB - A gradient elution high-performance liquid chromatographic method is described for the analysis of the beta-lactamase inhibitor tazobactam (YTR-830H) and a semi synthetic parenteral penicillin, piperacillin, in human plasma, serum, bile and urine. The assay for plasma, serum and bile involves deproteinization with acetonitrile and the removal of lipids with dichloromethane; urine is diluted with buffer. Separation and quantitation are achieved using a mobile phase based on ion-suppression chromatography on a C18 reversed-phase column with ultraviolet detection at 220 nm. The limit of quantitation for both compounds is 1.0 microgram/ml in plasma, serum and bile using a 0.2-ml sample and 50.0 micrograms/ml in urine using a 0.1-ml sample. The method has been validated by preparing and analyzing a series of fortified samples (range 1.0-200 micrograms/ml for each compound in plasma, serum and bile and 50.0-10,000 micrograms/ml for each compound in urine). Excellent linearity, accuracy, precision and recovery were obtained. The method was not interfered with by other endogenous components, nor by other commonly administered antibiotics such as amoxicillin, mezlocillin, cefometazole and cefotaxime. The assay has been successfully applied to the analysis of samples from pharmacokinetic studies in man and animals. PMID- 2556419 TI - Analysis of 3,4-diaminopyridine in human serum by solid-phase extraction and high performance liquid chromatography with ultraviolet detection. PMID- 2556420 TI - Thin-layer chromatography as an efficient alternative to high-performance liquid chromatography in the assay of plasma vitamin D3 metabolites. PMID- 2556421 TI - A method for determining the cut-off value of a varicella-zoster virus IgG enzyme immunoassay for immune status. AB - The cut-off optical density (OD) of an enzyme immunoassay (EIA) for IgG antibody to varicella-zoster virus (VZV) was established by application of a statistical technique to OD readings on sera from known susceptible and immune populations. Children aged one to three years who lacked complement fixation (CF) antibody were considered to be known susceptible subjects. Adults whose sera contained antibody by the CF test were considered to be known immune subjects. The method provides a valid alternative to previously used techniques of establishing a cut off OD, above which reading the EIA can be taken to indicate immunity to varicella-zoster infection. PMID- 2556423 TI - A plaque hybridization assay for rotaviruses. AB - A plaque hybridization assay was adapted to rotavirus. In this test cDNA or oligonucleotide probes were used to discriminate between plaques originating from virus carrying genes of bovine and simian origin. Only mRNAs present in infected cells were detected as demonstrated by using oligonucleotides corresponding to both strands. This assay can be used to screen reassortants or mutants of rotaviruses. PMID- 2556422 TI - Detection of human papillomavirus DNA in cervical cancer tissue by the polymerase chain reaction. AB - A method for detecting HPV DNA in cervical cancer tissue was developed without using isotopes. The DNA samples from the cancer tissues were first subjected to amplification by PCR, followed by polyacrylamide gel electrophoresis to identify the specific amplified fragment. The specificity and sensitivity of the PCR method are described. Compared with the dot hybridization technique, it is shown that the method is able to detect HPV DNA in cervical cancer tissues. PMID- 2556424 TI - Selective separation of virus proteins and double-stranded RNAs by SDS-KCl precipitation. AB - The total viral structural polypeptides and the double-stranded genomic RNAs of bluetongue virus can be selectively separated by a single SDS-KCl precipitation step. This simple, rapid and highly reproducible method enables greater than 95% recovery and purity of both viral proteins and dsRNAs within 30 min. The serotypic identity of the separated dsRNAs can be analyzed by SDS-PAGE electrophorogram immediately. After a single phenol/chloroform extraction, the dsRNA can also be used as hybridization probes, templates for molecular cloning and direct RNA sequencing. The SDS-KCl-precipitated viral proteins could be used readily for peptide mapping and as immunogens. Polyclonal and monoclonal antibodies raised against SDS-KCl-precipitated viral structural polypeptides were useful in Western immunoblots. PMID- 2556425 TI - Evaluation of end-point titration, single dilution and capture enzyme immunoassays for measurement of antirotaviral IgA and IgM in infantile secretions and serum. AB - In order to facilitate measurement of antirotaviral IgA in large collections of faeces and secretions, adaptations of enzyme immunoassay methods for estimating antirotaviral IgA and IgM in duodenal fluid, saliva, faeces and serum were studied. To quantitate specific IgA, a single dilution of each sample was assayed. Results were expressed as antirotaviral IgA units derived from a standard curve. Units were calculated by log-logit analysis on computer. There was strong correlation between antirotaviral IgA units and end-point titres in 257 faecal samples (correlation coefficient r = 0.92) and in 182 duodenal fluids and salivary samples (correlation coefficient r = 0.74). The assay was validated using acute and convalescent faeces from children with or without rotavirus infection. Immune conversions in IgA were detected in 33 (75%) of the children by units and 34 (77%) by titres. None of nine children with gastroenteritis due to other infectious agents showed immune conversions to rotavirus. A monoclonal capture IgM assay showed similar end-point titres and numbers of immune conversions when compared with a direct assay for antirotaviral IgM in serum and secretions. Use of the capture method eliminated false-positive reactions with the cell control. The assay for antirotaviral IgA units in secretions is simple, rapid, reproducible and reliable, and has proven of value in longitudinal epidemiological studies of rotavirus coproIgA profiles. Both the capture IgM technique and the single dilution IgA method permit analysis of large numbers of specimens and are appropriate for examination of immune responses to natural rotavirus infection or during vaccine trials. PMID- 2556426 TI - Discordant results obtained on testing sera from immunocompromised patients for cytomegalovirus IgG by enzyme-linked immunosorbent assay and radioimmunoassay. AB - The investigation of sera from immunocompromised patients for antibody to CMV by ELISA, RIA, immunofluorescence (IF) and complement-fixation (CF) revealed discrepancies that reflected differences in test specificity rather than sensitivity and suggested that for the long-term serological follow-up of such patients it would be advisable not to rely on only a single assay procedure. PMID- 2556427 TI - Hybridization techniques provide improved sensitivity for HCMV detection and allow quantitation of the virus in clinical samples. AB - Hybridization techniques (slot-blot and in-situ hybridization assays) and immunostaining using murine monoclonal antibodies directed against different proteins of the human cytomegalovirus (HCMV) were compared for their sensitivity and specificity for detection of HCMV. A model system with HCMV infected human embryonic lung fibroblasts and lung biopsy specimens obtained from patients with culture positive HCMV interstitial pneumonia were used for evaluation of these techniques. The hybridization techniques were found to provide an improved sensitivity compared to immunostaining. Additionally a good correlation was found between the virus dose determined by TCID50 and the amount of viral DNA detected by slot-blot hybridization and by the number of autoradiographic silver grains per 100 cells per 2 weeks exposure time detected in the infected fibroblasts by in-situ hybridization. Thus, at least in the model system quantification of the virus was achieved by hybridization assays. PMID- 2556428 TI - Polymerase chain reaction assay of parvovirus B19 DNA in clinical specimens. AB - The polymerase chain reaction (PCR) was used to detect parvovirus B19 DNA in a panel of sera from individuals recently infected with B19, as demonstrated by the presence of anti-B19 immunoglobulin M. Of 95 serum samples, 60 (63%) were found positive by PCR, whereas only 1 was also found positive by dot hybridization. In a control panel of 100 serum samples from individuals with other infections, only 1 serum sample was found positive by PCR, and this was also found positive by dot hybridization. This was probably just a fortuitous discovery of viremia. Placental tissues from women (n = 89) who had proven B19 infections in pregnancy but who gave birth to healthy infants at term were also tested. A total of 74 (83%) were found positive for B19 DNA by PCR. The high rate of detection by PCR probably represents "decay" of viral DNA after the peak of viremia and is not a clinically significant phenomenon. PMID- 2556429 TI - Amplification of human papillomavirus DNA sequences by using conserved primers. AB - The polymerase chain reaction has potential for use in the detection of small amounts of human papillomavirus (HPV) viral nucleic acids present in clinical specimens. However, new HPV types for which no probes exist would remain undetected by using type-specific primers for the polymerase chain reaction before hybridization. Primers corresponding to highly conserved HPV sequences may be useful for detecting low amounts of known HPV DNA as well as new HPV types. Here we analyze a pair of primers derived from conserved sequences within the E1 open reading frame for HPV sequence amplification by using the polymerase chain reaction. The longest perfect homology among HPV sequences is a 12-mer within the first exon of E1M. A region of conserved amino acids coded by the E1 open reading frame allowed the detection of another highly conserved region about 850 base pairs downstream. Two 21-mers derived from these conserved regions were used to amplify sequences from all HPV DNAs used as templates. The amplified DNA was shown to be specific for HPV sequences within the E1 open reading frame. DNA from HPVs whose sequences were not available were amplified by using these two primers. HPV DNA sequences in clinical specimens could also be amplified with the primers. PMID- 2556430 TI - Comparison of immunoblotting with other serological methods and virus isolation for the early detection of primary cytomegalovirus infection in allograft recipients. AB - Sequential specimens from nine allograft recipients were examined by using a variety of methods to detect primary cytomegalovirus (CMV) infection as rapidly as possible posttransplantation. Sera were examined for immunoglobulin G (IgG) and IgM antibodies by immunoblotting, enzyme immunoassay, and immunofluorescence and also by complement fixation, latex agglutination, and an immunofluorescence test for antibody to CMV early antigen. Urine and occasionally blood, tissue, and other specimens were centrifuged onto cell cultures to enhance CMV infectivity. Eight of the nine patients showed laboratory evidence of primary CMV infection, and CMV was isolated from seven of the eight: in no case was virus isolated before seroconversion had become evident. However, serological tests differed in their abilities to detect antibody response to CMV infection in different patients; while immunoblotting, latex agglutination, and enzyme immunoassay for IgG antibodies generally detected seroconversion before complement fixation, this was not invariably the case. At present, optimal laboratory detection of CMV infections in these patients can be achieved only by a combination of serological methods and virus isolation. PMID- 2556431 TI - Reactivity of VP4-specific monoclonal antibodies to a serotype 4 porcine rotavirus with distinct serotypes of human (symptomatic and asymptomatic) and animal rotaviruses. AB - Thirteen hybridomas secreting VP4-specific monoclonal antibodies against the Gottfried strain of porcine rotavirus (serotype 4) were produced and characterized. Nine of the hybridomas secreted neutralizing monoclonal antibodies (N-MAbs) against Gottfried rotavirus. These N-MAbs were divided into five distinct groups (groups I to V) according to their patterns of reactivity with different serotypes of human and animal rotaviruses. Group I N-MAbs (n = 3) were cross-reactive with five different serotypes of human rotavirus examined by a plaque reduction virus neutralization test. Group II N-MAbs (n = 3) neutralized all symptomatic human rotavirus serotypes tested and asymptomatic human rotavirus serotype 4 to a low titer. The single group III N-MAb neutralized mainly symptomatic human rotavirus serotypes 2 and 9 and none of the asymptomatic human rotavirus serotypes. The one N-MAb in group IV reacted at low titers with only asymptomatic human rotavirus serotypes 1 through 4. A group V N-MAb recognized serotype 4 porcine rotaviruses (Gottfried and SB-2) but no other human or animal rotaviruses examined. None of the N-MAbs recognized any animal rotaviruses tested (SA-11, RRV, OSU, NCDV, and B223), except for the Gottfried and SB-2 rotaviruses. The failure of N-MAbs (groups I to IV) to react with any animal rotaviruses tested but their ability to react variably with all human rotaviruses tested suggest that neutralizing epitopes on the VP4 protein are highly conserved between the Gottfried porcine and human rotaviruses. The Gottfried rotavirus may possibly represent a naturally occurring reassortant between pig and human rotaviruses or a rotavirus which is human in origin but pathogenic for swine. PMID- 2556432 TI - Thiol proteinase expression and pathogenicity of Entamoeba histolytica. AB - Expression of the 56-kilodalton (kDa) neutral thiol proteinase has been shown to correlate with the potential of clinical isolates of Entamoeba histolytica to produce invasive disease. A 56-kDa band was identified by gelatin substrate gel electrophoresis in 10 of 10 isolates from patients with colitis or amebic liver abscesses, but in only 1 of 10 isolates from asymptomatic patients. Pathogenic isolates appear capable of releasing significantly larger quantities of the proteinase, as measured by cleavage of a synthetic peptide substrate, ZRR-AMC (benzyloxy-carbonyl-arginine-arginine-4-amino-7-methylcoumarin). We have also shown that the proteinase is released during the course of clinical invasive amebic disease, as demonstrated by the presence of circulating antibodies detectable by enzyme-linked immunosorbent assay. These studies support the importance of the 56-kDa thiol proteinase in the pathogenesis of invasive amebiasis. PMID- 2556433 TI - Comparison of immunoglobulin A (IgA), IgG, and IgM enzyme-linked immunosorbent assays, plaque reduction neutralization assay, and complement fixation in detecting seroresponses to rotavirus vaccine candidates. AB - In a phase 1 study to evaluate human-rhesus rotavirus reassortant vaccines, 116 infants 1 to 5 months of age received one of the following five preparations: the serotype 1 reassortant, the serotype 2 reassortant, rhesus rotavirus (serotype 3), a bivalent preparation (serotypes 1 and 3), or a placebo. Seroresponses to the different vaccines were measured by plaque reduction neutralization assay (PRNA); rotavirus-specific immunoglobulin A (IgA), IgG, and IgM enzyme-linked immunosorbent assays (ELISAs); and complement fixation (CF). The seroresponse rate, calculated by using a fourfold or greater antibody rise by any assay, was similar in the four vaccine groups (83 to 96%). When the data from all the vaccinees were pooled, IgA ELISA, IgG ELISA, and PRNA were comparable in detecting seroresponses (67, 62, and 70%, respectively) and more efficient than IgM ELISA (53%) and CF (44%). When the vaccinees were analyzed by age, the overall seroresponse rates were the same for infants 1 to 2 and 3 to 5 months old (90%). The IgA ELISA and PRNA were the most efficient for detecting antibody rises in both age groups. IgG ELISA was among the least efficient methods for detecting antibody rises in the younger age group but among the most efficient in the older age group (44 versus 78%). CF was among the least efficient methods in both age groups but was significantly better in the older age group than in the younger age group (54 versus 21%). Our findings show that ELISA, in particular rotavirus-specific IgA ELISA, is a sensitive indicator of vaccine takes in 1- to 5 month-old infants, the target population for vaccination. ELISA should also be very useful in demonstrating natural rotavirus infections in field studies in which a stool specimen from a diarrheal episode is not always available. The ELISA has the advantages of being easier and quicker and requiring less serum than PRNA, but it does not give serotype-specific information about the immune response. PMID- 2556434 TI - Preparation of filamentous hemagglutinin from Bordetella pertussis and assay for serum antibodies to filamentous hemagglutinin and pertussis toxin for clinical and public health laboratories. AB - A procedure that is sufficiently simple and economical for use in clinical and public health laboratories for producing and purifying filamentous hemagglutinin (FHA) and determining antibodies to this major antigen of Bordetella pertussis in serum is described. High yields of FHA (40 to 80 mg/liter) were obtained in the supernatant by cultivating B. pertussis in modified CL medium. The FHA antigen was separated from pertussis toxin (PT) and other antigens by chromatography on hydroxylapatite. Removal of residual PT activity in the FHA fraction was effected by affinity absorption of PT with Fetuin immobilized to Sepharose 4B. The FHA was used as the antigen for determining titers of immunoglobulin G (IgG), IgA, and IgM to FHA in sera of patients with pertussis by an improved enzyme-linked immunosorbent assay. Development of the interfering background color commonly observed in conventional FHA enzyme-linked immunosorbent assay procedures was eliminated by washing the reaction wells with a buffer of high ionic strength before adding the peroxidase conjugates. In the absence of nonspecific background color, the reaction endpoints were easy to read. The FHA prepared by the procedure described was identical to a reference preparation of purified FHA in sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles and serological specificity assays. High yields of FHA were obtained from all four strains of B. pertussis tested in this study, indicating that the procedure for enhanced production of FHA may be generally applicable to other strains of B. pertussis. Results from tests of 50 serum specimens with clinical information on pertussis for FHA and PT antibodies by the assay procedures described exemplified the usefulness and caveats of serodiagnosis for pertussis. PMID- 2556437 TI - Comparative recovery of cytomegalovirus from saliva, mucolysed induced sputum, and bronchoalveolar lavage fluid from patients at risk for or with acquired immunodeficiency syndrome. AB - The recovery rates of cytomegalovirus from mucolysed induced sputum samples and bronchoalveolar lavage fluid obtained from individuals at risk for or with acquired immunodeficiency syndrome were compared. It was demonstrated that cytomegalovirus could be reliably recovered from mucolysed induced sputum, and such recovery was highly predictive of recovery from bronchoalveolar lavage samples obtained from the same individual. PMID- 2556435 TI - Temporal and geographical distributions of human rotavirus serotypes, 1983 to 1988. AB - Between 1983 and 1988, subgroups and serotypes were determined for 907 of 1,084 clinical specimens of rotaviruses collected in various countries of Europe, North and South America, Africa, and Asia. Enhanced enzyme immunoassays based on monoclonal antibodies specific for rotavirus proteins VP6 and VP7 were used. Significant differences in the prevalent serotypes were detected from year to year in the United Kingdom and Brazil and also in different countries during the same year. Throughout the study, rotavirus serotype 1 was detected most often (53.8%), followed in frequency by serotype 2 (17.8%), serotype 3 (12.1%), serotype 4 (11.1%), and serotypes other than 1 to 4 (5.1%). No individual serotype was found to predominate consistently in any one location. In the United Kingdom, rotavirus serotypes varied in prevalence in a regular but not predictable way. We suggest that a similar epidemiology might be found in other settings. Seventeen unusual strains were detected. Of these, five strains did not react with reference monoclonal antibodies specific for subgroup I and subgroup II, but they reacted with rotavirus group A-specific polyclonal and monoclonal antibodies; four strains were of subgroup II, serotype 2, and at least one had a "long" electropherotype; two strains were of subgroup I, serotype 2 with a long electropherotype; and one strain was of subgroup I, serotype 3. Five group C rotaviruses were detected. PMID- 2556436 TI - Comparison of monoclonal antibodies for rapid detection of cytomegalovirus in spin-amplified plate cultures. AB - Cytomegalovirus (CMV) was detected in 56 of 275 specimens (20%); 50 of 56 (89%) were detected by conventional culture, and 37 (66%) were detected by rapid assay at 72 h with a commercial monoclonal antibody and a pooled monoclonal antibody. Although the two antibodies were equally sensitive at 72 h, the pooled antibody gave a brighter, more easily detected signal. Other viruses were isolated from 9 specimens (3.3%) by conventional culture. Use of rapid assays alone fails to detect slow-growing CMV and non-CMV viral pathogens. PMID- 2556438 TI - Further antigenic characterization of porcine rotavirus YM. AB - We recently reported that the serotype specificity of YM, a group A porcine rotavirus, is different from those of the previously established rotavirus serotypes, 1 to 6 (A. M. Ruiz, I. V. Lopez, S. Lopez, R. T. Espejo, and C. F. Arias, J. Virol. 62:4331-4336, 1988). Here we demonstrate by cross-neutralization studies that rotavirus YM is also antigenically distinct from prototype rotavirus strains of serotypes 7 (Ty1), 8 (69M), 9 (WI61), and 10 (B223). A comparison of the serotype-determining amino acid regions of the YM surface glycoprotein, VP7, with the reported homologous sequences of rotavirus strains Gottfried, 69M, B37, F45, and WI61 further supported the serotypic individuality of the rotavirus YM VP7 protein. PMID- 2556439 TI - Evaluation of a latex particle agglutination assay for the detection of cytomegalovirus antibody in patient serum. AB - The Virogen CMV Antibody Test is a simple and rapid latex agglutination assay for the detection of cytomegalovirus antibody in human serum and plasma. Evaluation of this assay with respect to enzyme immunoassay yielded a sensitivity of 98% with a specificity of 100%. In comparison to CMVScan, the Virogen CMV Antibody Test had a sensitivity of 98.4% and a specificity of 100%. PMID- 2556440 TI - Continuous high-speed rolling versus centrifugation for detection of herpes simplex virus. AB - Specimens submitted for diagnosis of herpes simplex virus infections were inoculated into shell vials and conventional culture tubes. Inoculated culture tubes were incubated with rolling at 96 rpm. Immunoperoxidase (IP) staining and cytopathic effects (CPE) were used to detect positive cultures. At 24 h, 42 (53%) of the rolled cultures were positive for CPE, while only 16 (21%) of the shell vials were CPE positive (P less than 0.01). No difference in sensitivity was seen between rolled and shell vial cultures that were inoculated with high-titered viral preparations and IP stained at 16 h. However, when low-titered preparations were used, 39 of 41 (95%) were IP positive by the high-speed roller method at 64 h postinoculation, while only 24 of 41 (58%) were IP positive with shell vials (P less than 0.01). These results indicate that high-speed roller method at 64 h postinoculation, while only 24 of 41 (58%) were IP positive with shell vials (P less than 0.01). These results indicate that high-speed rolling is better than the shell vial technique for the detection of herpes simplex virus by IP staining. PMID- 2556441 TI - Effects of tetracaine and procaine on skinned muscle fibres depend on free calcium. AB - The local anaesthetics, tetracaine and procaine have previously been found to block, induce or potentiate Ca2+ release from the sarcoplasmic reticulum (SR) of skeletal muscle depending on the preparation, experimental conditions and design. We now show that low concentrations of tetracaine and procaine block SR Ca2+ release whereas high concentrations induce release from the SR of amphibian and mammalian skinned fibres. Both actions depend on pCa, such that a shift in pCa can alter their effect from blocking to releasing Ca2+. In skinned fibres with Ca2+-loaded SR, tetracaine (1 mM) produced a tonic contraction with a time to half-peak of 15-20 s and a magnitude reaching 80% of maximum force. Ca2+ release by tetracaine or procaine occurred at pCa less than or equal to 6.5 and was not blocked by Ruthenium Red (RR) (25 mM). This action of tetracaine was attributed to SR Ca2+ release rather than to a displacement of bound Ca2+ because fibres lacking a functional SR due to pre-treatment with quercetin (100 mM), A 23187 (100 micrograms ml-1) or Triton X-100 (1%) did not contract after additions of tetracaine. Lower concentrations of tetracaine (0.5 mM) and procaine (less than or equal to 10 mM) blocked contractions due to caffeine (at pCa greater than or equal to 6.73), sulphydryl oxidizing agents, or Ca2+-induced Ca2+ release (CICR). The inhibition of CICR as a function of pCa was difficult to measure quantitatively since lowering pCa to elicit CICR twitches was sufficient to initiate tetracaine-induced tonic contractions. Experiments with isolated SR vesicles showed that 1 mM tetracaine inhibited CICR, over a wide range of pCa but 3-5 mM tetracaine induced rapid Ca2+ release. The opposite effects of tetracaine and procaine depend mostly on their concentration in SR vesicles and/or pCa in skinned fibres. Blockade of release seems to occur via the CICR pathway, and induction of release through an increase in SR membrane permeability. PMID- 2556442 TI - Regulation of interleukin 3 gene induction in normal human T cells. AB - The regulation of IL-3 gene induction in human peripheral blood T cells was studied. IL-3 gene expression was inducible by crosslinking of the T cell receptor/CD3 complex using anti-CD3 MAb G19-4. Anti-CD3-induced IL-3 gene expression was found to be limited to the CD28+ T cell subset and could be augmented by costimulating T lymphocytes with antibodies directed against CD28. IL-3 expression could also be induced by costimulation of T cells with both phorbol ester and ionomycin, which are thought to mimic the intracellular effects of T cell receptor-antigen interaction. However, unlike other lymphokines such as IL-2 or granulocyte-macrophage colony-stimulating factor, IL-3 gene expression is not induced by stimulation of cells with phorbol myristate acetate and anti-CD28. We conclude that IL-3 gene regulation is under stringent control since IL-3 gene expression occurs only in the CD28+ subset of T cells, and since IL-3 induction obligately requires increased intracellular calcium. PMID- 2556443 TI - Chronic norepinephrine elicits desensitization by uncoupling the beta-receptor. AB - The goal of this study was to determine the mechanism of beta-adrenergic receptor desensitization after chronic elevation of circulating NE levels. Osmotic minipumps containing either NE or saline were implanted subcutaneously in dogs for 3-4 wk. Physiologic desensitization to isoproterenol was confirmed in conscious dogs, i.e., left ventricular dP/dt increased in response to isoproterenol (0.4 micrograms/kg per min) by 5,625 +/- 731 mmHg/s in control dogs with saline pumps, and significantly less, P less than 0.01, by 2,093 +/- 263 mmHg/s in dogs with NE pumps. Myocardial beta-adrenergic receptor density as determined with 125I-cyanopindolol binding was 49% higher (p less than 0.05) in the NE pump group. However, beta-adrenergic receptor agonist binding with isoproterenol demonstrated a significant shift into the low affinity state for the animals with NE pumps. Basal, GTP plus isoproterenol, 5' guanylylimidodiphosphate, sodium fluoride, and forskolin-stimulated adenylate cyclase activity in the NE pump group were significantly depressed (P less than 0.05) by amounts ranging from 20 to 40%. The functional activity of the guanine nucleotide binding protein Gs was also reduced (P less than 0.05) in animals with NE pumps. Thus, the process of desensitization in response to chronic elevation of NE levels in intact, normal dogs does not involve a decrease in beta adrenergic receptor density. Rather, it is characterized by reduced adenylate cyclase activation and uncoupling of the beta-adrenergic receptor in association with decreased activity of the GTP-coupling protein Gs. PMID- 2556444 TI - An arginine to glutamine mutation in residue 109 of human ornithine transcarbamylase completely abolishes enzymatic activity in Cos1 cells. AB - Ornithine transcarbamylase (OTC) is an important enzyme in the detoxification of ammonia to urea, and its deficiency is the most common inborn error of ureagenesis in humans. Among 24 cases of OTC deficiency previously examined, three unrelated individuals all showed loss of a Taq I site in the OTC gene corresponding to codon 109, suggesting that this Taq I site may be prone to mutation. Two of these patients demonstrated the same C----T transition (in antisense strand) converting Arg109 to Gln. Although these studies implied a strong association between the missense mutation and OTC-deficient phenotype, a causal relationship could not be firmly established. We have investigated this relationship by reconstructing the mutation in vitro. A full-length human OTC cDNA was cloned into an SV40-based expression vector and has been reproducibly expressed at high levels in the cell line Cos1. By site-directed mutagenesis of this wild type sequence, we constructed a missense mutation which contains the C- --T transition. Electroporation and transient assay in Cos1 indicated that the specific activity of mutant OTC was 100-fold lower than that of wild type. This result confirms that the Taq I alteration leading to the Gln missense is responsible for the OTC deficiency affecting the above patients. PMID- 2556445 TI - Activation of cultured rat hepatic lipocytes by Kupffer cell conditioned medium. Direct enhancement of matrix synthesis and stimulation of cell proliferation via induction of platelet-derived growth factor receptors. AB - Hepatic lipocytes appear to be central to the pathogenesis of hepatic fibrosis, undergoing activation during inflammation to a matrix-producing, proliferative cell type. We have studied the activation process in culture by examining the response of lipocytes to conditioned medium from hepatic macrophages (Kupffer cells). Lipocytes exposed to Kupffer cell medium (KCM) exhibited cellular and nuclear enlargement associated with up to a threefold increase in collagen and total protein synthesis per cell. Cell proliferation was also stimulated as measured by [3H]thymidine incorporation and direct cell counting. The latter effect was serum dependent and inhibited by antibodies to platelet-derived growth factor (PDGF). Proliferation could be stimulated by recombinant PDGF, but only after preincubation of cells with KCM. These findings suggested that KCM was eliciting expression of the PDGF receptor in lipocytes, and this was confirmed by immunoblot analysis with antibodies to the PDGF receptor. DNA synthesis in lipocytes exposed to KCM occurred at 48 h, which reflected the time required for PDGF receptor expression (24 h) plus initiation of [3H]thymidine incorporation (24 h). These results indicate that KCM has multiple stimulatory effects on cultured lipocytes similar to activation of these cells observed in vivo. PMID- 2556446 TI - Cytochrome P450 metabolites of arachidonic acid are potent inhibitors of vasopressin action on rabbit cortical collecting duct. AB - AA is metabolized by a cytochrome P450, NADPH-dependent epoxygenase to four regioisomeric epoxyeicosatrienoic acids (EETs). The EETs are further hydrated enzymatically to their respective diols, vic-dihydroxyeicosatrienoic acids (DHETs). We studied the effect of pretreatment with DHETs on 10 microU/cm2 arginine vasopressin (AVP)-stimulated hydraulic conductivity (Lp) (Lp x 10(-7) cm/atm/s, mean +/- SE) in rabbit cortical collecting ducts (CCDs) perfused in vitro at 37 degrees C. At 10(-6) M all four DHETs were potent inhibitors of the hydroosmotic effect of AVP. 14,15-DHET was the most potent isomer; it reduced AVP induced Lp from a control value of 234.75 +/- 11.7, n = 17, to a value of 95.2 +/ 8.39, n = 5, P less than 0.0001, a reduction of AVP-mediated water flow of 60%. The inhibitory effect of 14,15-DHET was dose dependent and significant to nanomolar concentrations. 14,15-DHET at 10(-7) M was as potent an inhibitor of AVP's activity as was 10(-7) M PGE2. AVP's hydroosmotic effect is mediated through its intracellular second messenger, cAMP. 8-p-Chlorophenylthio-cAMP (CcAMP) at 10(-4) M induced a peak Lp of 189.6 +/- 11.0, n = 8; pretreatment with 10(-6) M 14,15-DHET reduced CcAMP-peak Lp to 132.0 +/- 13.4, n = 5, P less than 0.01, demonstrating a post-cAMP effect. Gas chromatography/mass spectroscopy suggests that EETs are present in extracts purified from CCDs. We conclude that cytochrome P450 epoxygenase eicosanoids are potent inhibitors of the hydroosmotic effect of vasopressin and are endogenous constituents of normal CCDs, the major target tissue for AVP. PMID- 2556447 TI - Macrophage/monocyte receptor for nonenzymatically glycosylated protein is upregulated by cachectin/tumor necrosis factor. AB - Proteins of extracellular matrix undergo over time multiple reactions with glucose to form advanced glycosylation endproducts (AGEs) which are highly active in protein crosslinking, and have been implicated in tissue damage associated with aging and diabetes. A macrophage/monocyte receptor for AGE moieties mediates the uptake of AGE-modified proteins by a process that also induces cachectin/tumor necrosis factor (TNF) and IL-1 secretion. Reasoning that cytokines might regulate this AGE-receptor system, we have evaluated the effect of cachectin/TNF, IL-1, and IFN-gamma on AGE-protein processing. We report that cachectin/TNF induced a severalfold enhancement of binding, endocytosis, and degradation of AGE-BSA by both murine peritoneal macrophages and human blood monocytes in vitro, and that cachectin/TNF enhanced the rate of disappearance of AGE-modified red blood cells in vivo. IL-1 and IFN-gamma alone did not increase AGE processing, but IFN-gamma consistently enhanced cachectin/TNF-induced changes in AGE-receptor kinetics. Similar effects were induced by AGE-BSA and FFI-BSA, a chemically synthesized AGE, when used as macrophage stimulants, possibly via cachectin/TNF induction. All upregulatory responses were blocked by anticachectin/TNF monoclonal antibody. These data suggest that AGE-induced cachectin/TNF, in addition to influencing tissue regeneration and remodelling, may also normally regulate the disposal of tissue damaging AGE-proteins through an autocrine upregulation. PMID- 2556449 TI - Localization of integrin receptors for fibronectin, collagen, and laminin in human skin. Variable expression in basal and squamous cell carcinomas. AB - VLA integrins in human skin were examined by indirect immunofluorescence utilizing antibodies recognizing the beta 1, alpha 2, alpha 3, or alpha 5 subunits. Staining of fetal, newborn, or adult skin with antibodies to beta 1, alpha 2, or alpha 3 subunits gave essentially similar staining patterns: intense staining was associated with the basal layer of the epidermis, hair follicles, and blood vessel walls. The alpha 5 subunit could be detected only in epidermis and the inner root sheath of hair follicles in fetal skin. In epidermis, the staining reaction for the beta 1 subunit was not only found in sites interfacing with the basement membrane zone, but also around the entire periphery of these cells. We speculate that these receptors might have previously unrecognized functions in cell-cell interactions or that these findings may suggest the presence of previously unrecognized ligands in the intercellular spaces of keratinocytes. Examination of nine nodular basal cell carcinomas revealed a prominent staining reaction with anti-beta 1 and anti-alpha 3 antibodies at the periphery of the tumor islands. In contrast, staining of five squamous cell carcinomas revealed either the absence of integrins or altered and variable expression. Thus, matrix components and their receptors may participate in modulation of growth, development, and organization of human skin. PMID- 2556448 TI - High receptor binding affinity of lipoproteins in atypical dysbetalipoproteinemia (type III hyperlipoproteinemia). AB - Familial dysbetalipoproteinemia (or type III hyperlipoproteinemia) is characterized by the presence of abnormal, cholesteryl ester-rich beta-very low density lipoproteins (beta-VLDL) in the plasma. Subjects with typical dysbetalipoproteinemia are homozygous for an amino acid substitution in apolipoprotein (apo-) E at residue 158 and have defective apo-E-mediated binding of both pre-beta-VLDL and beta-VLDL to apo-B,E(LDL) (or LDL) receptors (1988. Chappell, D.A., J. Clin. Invest. 82:628-639). To understand the effect of substitutions in apo-E at sites other than residue 158, nine dysbetalipoproteinemic (dys-beta) subjects who were either homozygous or heterozygous for substitutions in apo-E at atypical sites were studied. These substitutions occurred at residue 142 (n = 6), 145 (n = 2), or 146 (n = 1) and are known to cause less defective binding than does the 158 substitution. The chemical composition and electrophoretic mobility of pre-beta-VLDL and beta-VLDL from atypical and typical dys-beta subjects were indistinguishable. However, lipoproteins from atypical and typical dys-beta subjects differed in their affinity for the apo-B,E(LDL) receptor on cultured human fibroblasts. The pre beta-VLDL and beta-VLDL from atypical dys-beta subjects had 640- or 17-fold higher affinity, respectively, than did corresponding lipoproteins from typical dys-beta subjects. The higher binding affinity of lipoproteins from atypical dys beta subjects was associated with a higher ratio of apo-E to total apo-C. Since higher binding affinity should cause more rapid receptor-mediated clearance of beta-VLDL in atypical than in typical dys-beta subjects in vivo, the mechanism of beta-VLDL accumulation may differ in these two groups. PMID- 2556451 TI - Placental corticotropin-releasing hormone may be a stimulator of maternal pituitary adrenocorticotropic hormone secretion in humans. AB - To clarify the physiological role of placental corticotropin-releasing hormone (CRH), we measured plasma CRH, ACTH, and cortisol throughout pregnancy. Cerebrospinal fluid (CSF) CRH levels and ACTH responsiveness to synthetic CRH were also quantified in pregnant and nonpregnant women. Maternal plasma CRH levels, which increased progressively during pregnancy, correlated well with both ACTH and cortisol in early labor, delivery, and postpartum samples, and also with cortisol levels in samples before labor. CSF CRH levels in term pregnant women did not differ from those of nonpregnant women. CRH infusion that attained similar plasma CRH levels to those found in late pregnancy elicited significant ACTH release in vivo and regular CRH test provoked normal ACTH response during early pregnancy but no response during late pregnancy. We concluded that: (a) maternal pituitary-adrenal axis correlates well with plasma CRH levels, which are high enough to provoke ACTH release from maternal pituitary; (b) hypothalamic CRH secretion in term pregnant women is not exaggerated; and (c) maternal pituitary is responsive to synthetic CRH in early but not late pregnancy, suggesting that maternal pituitary-adrenal axis is already activated by high circulating CRH. Placental CRH may be an important stimulator of the maternal pituitary-adrenal axis during pregnancy. PMID- 2556450 TI - Identification and characterization of a monocyte-derived neutrophil-activating factor in corticosteroid-resistant bronchial asthma. AB - Peripheral blood mononuclear cells (PBMC) were isolated from seven normal subjects, eight asthmatic subjects clinically sensitive to corticosteroids (CS), and eight asthmatic subjects clinically resistant to corticosteroids (CR). PBMC were cultured at 37 degrees C for 24 h in the absence or presence of 10(-16) to 10(-4) M hydrocortisone. Calcium ionophore (A23187)-activated neutrophils (PMN) primed by supernatants of PBMC from asthmatic subjects cultured in the absence of hydrocortisone generated approximately threefold more leukotriene B4 than PMN primed by supernatants of PBMC from normal subjects (P less than 0.05). Incubation of PBMC derived from CS subjects with 10(-8) M hydrocortisone completely inhibited the production of the enhancing activity (P less than 0.01), whereas in CR subjects hydrocortisone at concentrations up to 10(-4) M did not suppress the release of enhancing activity. The enhancing activity was produced by monocytes. Enhancing activity eluted with an Mr of 3,000 D and a pI of 7.1. It eluted at 10% acetonitrile after reverse-phase HPLC. The activity was destroyed by heating to 60 degrees C for 60 min and was sensitive to pronase treatment. The purified factor also enhanced superoxide generation by PMN which had been stimulated submaximally by phorbol myristate acetate. PMID- 2556454 TI - Plasma Ip(a) concentration is inversely correlated with the ratio of Kringle IV/Kringle V encoding domains in the apo(a) gene. AB - Plasma Lp(a) levels correlate with atherosclerosis susceptibility. This lipoprotein consists of an LDL-like particle attached to a large glycoprotein called apo(a). Apo(a) is a complex glycoprotein containing multiple Kringle domains, found to be highly homologous to plasminogen Kringle IV, and a single Kringle domain homologous to plasminogen Kringle V. Lp(a) levels appear to be inversely correlated with apo(a) size in a given individual. In this study, we have used probes specific to the Kringles IV and V domains of apo(a) cDNA in quantitative Southern blotting analysis. By this method, we have determined the ratio of Kringle IV/Kringle V encoding domains in the apo(a) gene of 53 unrelated individuals with different plasma concentrations of Lp(a). This ratio was found to be inversely correlated with log Lp(a) levels (r = -0.90, P less than 0.0001) and directly correlated with apo(a) apparent molecular weight (Mr) (r = 0.79, P less than 0.0001). In summary, by showing that Lp(a) concentrations and apo(a) apparent size are highly correlated with the ratio of Kringle IV/Kringle V encoding domains in the apo(a) gene, we provide a DNA marker for this atherosclerosis risk factor as well as an important insight into the genetic mechanism regulating Lp(a) levels. PMID- 2556455 TI - Salivary duct carcinoma in major and minor salivary glands. PMID- 2556453 TI - A missense mutation in the neutrophil cytochrome b heavy chain in cytochrome positive X-linked chronic granulomatous disease. AB - A membrane-bound cytochrome b, a heterodimer formed by a 91-kD glycoprotein and a 22-kD polypeptide, is a critical component of the phagocyte NADPH-oxidase responsible for the generation of superoxide anion. Mutations in the gene for the 91-kD chain of this cytochrome result in the X-linked form of chronic granulomatous disease (CGD), in which phagocytes are unable to produce superoxide. Typically, there is a marked deficiency of the 91-kD subunit and the cytochrome spectrum is absent (X- CGD). In a variant form of CGD with X-linked inheritance, affected males have a normal visible absorbance spectrum of cytochrome b, yet fail to generate superoxide (X+ CGD). The size and abundance of the mRNA for the 91-kD subunit and its encoded protein were examined and appeared normal. To search for a putative mutation in the coding sequence of the 91-kD subunit gene, the corresponding RNA from an affected X+ male was amplified by the polymerase chain reaction and sequenced. A single nucleotide change, a C----A transversion, was identified that predicts a nonconservative Pro----His substitution at residue 415 of the encoded protein. Hybridization of amplified genomic DNA with allele-specific oligonucleotide probes demonstrated the mutation to be specific to affected X+ males and the carrier state. These results strengthen the concept that all X-linked CGD relates to mutations affecting the expression or structure of the 91-kD cytochrome b subunit. The mechanism by which the Pro 415----His mutation renders the oxidase nonfunctional is unknown, but may involve an impaired interaction with other components of the oxidase. PMID- 2556452 TI - Activation of normal and cystic fibrosis Cl- channels by voltage, temperature, and trypsin. AB - In cystic fibrosis (CF) phosphorylation-dependent activation of outwardly rectifying apical membrane Cl- channels is defective. To further understand regulation of this channel we examined several other mechanisms of channel activation in normal and CF cells. Previous studies have shown that strong membrane depolarization can activate channels in excised cell-free membrane patches. Here we show that such activation is dependent on both the absolute membrane voltage and the duration of depolarization. Moreover, activation was reversible by membrane hyperpolarization. In some cases, excising patches of membrane from the cell caused channel activation, even in the absence of depolarization. However, the frequency of channel activation with patch excision increased when bath temperature was increased from 23 to 37 degrees C. Although the channel remained in the activated state when temperature was reduced to 23 degrees C, subsequent hyperpolarization inactivated the channel. In cell-attached patches, neither depolarization nor increasing bath temperature to 37 degrees C activated channels, suggesting that neither is physiologically important in regulation of the channel. Thus changes in membrane voltage and bath temperature appear to cause a nonenzymatic change in the channel's conformation; the interactions between voltage and temperature suggest that they may affect the same process. To determine if a proteolytic alteration of the channel could also cause activation, we added trypsin to the cytosolic surface of excised membrane patches. Trypsin activated channels, which could not then be inactivated by either hyperpolarization or phosphorylation with PKC, suggesting that trypsin removed or altered a region of the channel involved in inactivation. All of these interventions activated Cl- channels from both normal and CF cells. Thus many aspects of Cl- channel activation are normal in CF; only phosphorylation dependent activation is defective. PMID- 2556456 TI - Potassium homeostasis during angiotensin-converting enzyme inhibition with enalapril. AB - The effect of angiotensin-converting enzyme (ACE) inhibition on renal and extrarenal potassium (K) regulation was examined. Six healthy men were studied in double-blinded crossover fashion on placebo or enalapril, 80 mg/day. On day 4, the subjects were given an intravenous infusion of KCl and on day 5 an oral dose of 10% NH4Cl. Treatment with enalapril decreased plasma aldosterone and increased plasma renin activity (PRA), epinephrine and norepinephrine, but did not affect serum glucose, plasma insulin or basal plasma K. Maximal increases in plasma K during K infusion or NH4Cl ingestion were similar during enalapril and placebo treatment. With enalapril treatment urinary K excretion was unchanged following K loading but moderately reduced following NH4Cl loading. We conclude that ACE inhibition does not acutely impair K homeostasis in men with normal renal function. PMID- 2556457 TI - Distribution of beta-nerve growth factor receptors in the human basal forebrain. AB - The distribution of neurons expressing the receptor for beta-nerve growth factor has been examined immunohistochemically in serial coronal sections of basal forebrain from aged normal human subjects. Neurons expressing the receptor were observed in the nucleus of the diagonal band of Broca and in the anterior, the intermediate, and the posterior portions of the nucleus basalis of Meynert. Neurons could also be seen in the medial septal nucleus and embedded in myelinated fibre tracts such as those of the external capsule, cingulum, medullary laminae of the globus pallidus, ansa penduncularis, ansa lenticularis, and anterior commissure. In situ hybridization with a 35S cDNA probe to the human beta-nerve growth factor receptor confirms a neuronal location as the site of synthesis of beta-nerve growth factor receptors in the nucleus basalis of Meynert in a fifth brain. A high percentage of Nissl-stained hyperchromic magnocellular neurons expressed the receptor for beta-nerve growth factor, suggesting that most neurons in the human cholinergic magnocellular basal forebrain system express these receptors. Recent data suggest that beta-nerve growth factor functions as a neurotrophic factor in basal forebrain cholinergic neurons. In Alzheimer's disease there is known to be a reduction in cholinergic function and an apparent loss of neurons in the cholinergic nucleus basalis of Meynert. For this reason we have examined the distribution of receptors for beta-nerve growth factor in the normal human basal forebrain in order to form a basis for comparison to those with Alzheimer's disease. PMID- 2556458 TI - Chondroid syringoma associated with hidrocystoma-like changes. Possible differentiation into eccrine gland. A histologic, immunohistochemical and electron microscopic study. AB - A case of chondroid syringoma associated with hidrocystoma-like changes was investigated by histology, immunohistochemistry and electron microscopy. Chondroid syringoma was histologically compatible with apocrine mixed tumor, and hidrocystoma-like changes did not fulfill diagnostic criteria of either eccrine hidrocystoma or apocrine hidrocystoma. However, epithelial cellular elements composing both chondroid syringoma and hidrocystoma-like changes suggested, immunohistochemically and electron microscopically, differentiation into eccrine gland. The lesions of both had an apparent transition of ductal structures of chondroid syringoma into hidrocystoma-like changes. Therefore, chondroid syringoma and hidrocystoma-like changes in this case may be organized as a peculiar type of cutaneous appendage tumor differentiating toward eccrine gland. PMID- 2556459 TI - Effects of dietary neutral detergent fiber concentration and supplementary long hay on chewing activities and milk production of dairy cows. AB - Effects of dietary NDF concentration on chewing and productivity were assessed using silage-based diets with and without supplemental long hay. Twelve Holstein cows (125 d postpartum) were used in a double 6 x 6 Latin square to evaluate six diets formulated using high moisture shelled corn and alfalfa silage (37% DM, 23% CP, 48% NDF) to provide three concentrations of NDF: 26, 30, and 34%. At each concentration, an alternative diet was formulated by substituting 15% of the silage DM with an equivalent amount of long alfalfa grass hay (14% CP, 61% NDF). Cows were fed at 85% of ad libitum intake, and ingredients were allocated separately. Increasing NDF decreased milk yield from 20.8 to 19.9 and 19.1 kg/d, for 26, 30, and 34%, respectively. Supplementing diets with hay increased milk production by .7 kg/d, although milk fat content was not affected. Increasing NDF resulted in a quadratic increase in ruminating and total chewing time from 344 and 558 for 26% NDF, to 413 and 651 for 30%, and 414 and 674 min/d for 34%, respectively. Added hay did not increase daily ruminating and chewing time; ruminating time per unit of NDF intake was reduced by hay supplementation (75.3 vs. 69.4 min/kg). PMID- 2556460 TI - Utilization of three maturities of alfalfa by dairy cows fed rations that contain similar concentrations of fiber. AB - The effect of alfalfa maturity on ration utilization and lactation performance by high producing dairy cows was measured in a 13-wk lactation study. Eighteen multiparous Holsteins were fed one of three rations containing first crop alfalfa hay harvested at early vegetative (36.1% NDF), late bud (51.7% NDF), or full bloom (51.7% NDF) maturity. Forage to concentrate ratios were 68:32, 53:47, and 45:55 for early vegetative, late bud, and full bloom rations, respectively. As offered, the late bud ration was higher in fiber (34.6% NDF) than either the early vegetative (31.8% NDF) or full bloom (30.6% NDF) rations. Cows fed the ration with early vegetative hay produced as much 4% FCM (32.5 kg) as cows fed the ration containing more fiber (late bud, 32.9 kg) or the later maturity hay (full bloom, 32.2 kg). Milk fat and protein composition were not affected by ration fiber concentration or forage maturity. Average dry matter intake per day and body weight change were similar across treatments. Time spent ruminating was similar for the three rations (6.8 h/d), but eating time was affected by forage maturity (early vegetative, 4.4 h/d vs. full bloom, 5.7 h/d). Cows receiving the higher fiber diet (late bud, 6.2 h/d) spent more time eating than cows receiving the lower fiber diets. Fiber concentration and forage maturity did not affect milk production, milk composition, or body weight change of high producing dairy cows in early lactation. PMID- 2556461 TI - Effect of source of neutral detergent fiber and starch on nutrient utilization by dairy cows. AB - Two diets were formulated to contain 30% NDF (DM basis) but differed in source of NDF and type of concentrate. One diet contained approximately 70% alfalfa silage and 30% corn grain (89% of NDF from forage) and the other diet contained about 50% alfalfa silage and 50% barley grain (65% of NDF from forage). Diets were fed to 12 cows in a short production trial, a total collection digestion experiment, and a rate of passage study. Cows fed the alfalfa-corn diet produced more 4% FCM (22.4 vs. 20.7 kg/d) than did cows fed the alfalfa-barley diet. Intake of DM (3.7% of BW), NE1 (35 Mcal), and NDF (1.1% of BW) were similar between diets, but due to differences in diet composition, intake of ADF was greater by cows fed alfalfa-corn than by cows fed alfalfa-barley. Digestibilities of DM, NDF, and CP were not affected by diet, but hemicellulose digestion was higher and cellulose and ADF digestibilities were lower by cows fed alfalfa-barley diets. Cows fed alfalfa-corn diet had a higher ruminal acetate to propionate molar ratio. No effect of diet was observed on turnover kinetics of the forage, concentrate, or liquid markers. These data show that forage to grain ratio and source of starch must be considered prior to recommending that diets be balanced for NDF. PMID- 2556462 TI - Dietary interaction of cane molasses with source of roughage: intake and lactation effects. AB - Effects of cane molasses at 0, 4, and 8% of DM in complete mixed diets were evaluated when molasses was fed to lactating dairy cows with cottonseed hulls, alfalfa haylage, or both combined. Thirty-six mature Holstein cows from mid to late lactation were used in a partially balanced incomplete block design with three 28-d periods. Variables measured were DM intake, milk yield, and composition. With cottonseed hull diets (30% of DM), molasses improved milk yield, milk fat percentage, solids-corrected milk, and feed efficiency. Dry matter intake was not affected by molasses. With alfalfa haylage diets (35% of DM), 8% molasses depressed actual milk yield and solids-corrected milk, DM intake, milk fat percentage, milk protein percentage, and feed efficiency. Milk fat percentage was increased with 4% molasses. With 65% alfalfa haylage diets, most variables measured were unaffected by molasses; however, with 8% molasses, DM intake increased and milk protein percentage decreased. In diets with both roughages combined, molasses did not affect any variable measured; however, these diets gave highest solids-corrected milk yields. Results show clearly that molasses effects depend on percentage molasses and type and amount of roughage in the diet. PMID- 2556463 TI - Stump the experts. Eccrine acrospiroma. PMID- 2556464 TI - Treatment of malignant fibrous histiocytoma and atypical fibrous xanthomas with micrographic surgery. AB - Fibrous tumors of the soft tissue are usually benign, but some fibrous neoplasms such as dermatofibrosarcoma protuberans (DFSP), atypical fibroxanthoma (AFX), and malignant fibrohistiocytoma (MFH) can be very destructive locally with a high recurrence rate after local excision. On occasion, they can metastasize. Previous reports have confirmed the high success rate of Mohs micrographic surgery for the treatment of DFSP, but data have been lacking on the potential benefit of this surgical approach for MFH and AFX tumors. Over the past 6 years, we have treated 17 patients with MFH (20 tumors) and 5 patients with AFX with Mohs micrographic surgery. A retrospective analysis of the surgical results is presented. To date (average 3-year follow-up), all patients contacted are tumor free with only one recurrence; no patient has developed metastatic disease. Our results to date are very encouraging; they lend support to Mohs micrographic surgery as a desired surgical approach for these difficult-to-cure neoplasms. PMID- 2556465 TI - Mistaken injection of sodium bicarbonate. PMID- 2556466 TI - Recurrent herpes simplex virus and the acceleration of the wasting syndrome: report of case. PMID- 2556467 TI - Dentistry on stamps. PMID- 2556468 TI - Relative metabolism of quinones to semiquinone radicals in xanthine oxidase system. AB - Rates of enzymatic single-electron reduction of some myotoxic quinones to semiquinone metabolites in an in vitro xanthine oxidase/hypoxanthine/catalase system varied widely. Naphthoquinones, especially juglone, were found to undergo rapid single-electron reduction. Benzoquinones and benzoquinoneimines, as well as phenanthrene-9,10-quinone, benzo[a]pyrene-3,6-quinone, and diethylstilbestrolquinone, were also actively reduced. The anthraquinones danthron, doxorubicin and emodin were poorly metabolized in this system. N Acetylcysteine inhibited quinone-stimulated cytochrome C reduction at high concentrations. The results of this study are discussed with respect to cytotoxicity and mutagenicity of selected quinones. PMID- 2556470 TI - Effects of cortisol or corticotropin administration on hepatic 3-hydroxy-3 methylglutaryl-CoA reductase activity and plasma lipids in the pregnant rat and fetuses. AB - Pregnant rats were given pharmacological doses of cortisol or ACTH or no hormone from gestation day 9 to 19 and maternal and fetal hepatic 3-hydroxy-3 methylglutaryl-CoA reductase activity and plasma cholesterol studied on gestation day 20. Reductase activity was also studied in the maternal and fetal adrenal of the rats given cortisol or no hormone. Cortisol administration increased the maternal and fetal plasma cholesterol but had no effect on the hepatic active (phosphorylated) 3-hydroxy-3-methylglutaryl-CoA reductase activity when compared to untreated rats. Total (active + inactive) 3-hydroxy-3-methylglutaryl-CoA reductase activity, however, was reduced in maternal liver but not altered in the fetal liver by cortisol. The maternal cortisol treatment decreased the fetal, but not maternal, adrenal 3-hydroxy-3-methylglutaryl-CoA reductase total enzyme activity. The data support a hypothesis that utilization of plasma cholesterol for adrenal steroidogenesis may be an important determinant of plasma cholesterol homeostasis in the rat fetus. Maternal ACTH administration increased the foetal but not maternal plasma cholesterol, whilst active 3-hydroxy-3-methylglutaryl-CoA reductase activity was increased in the pregnant rat but not her fetuses. This result may suggest coordination of hepatic active reductase activity with adrenal cholesterol utilization in the pregnant rat. The reason for the fetal hypercholesterolaemia caused by ACTH, which is not known to cross the placenta, is uncertain. The studies, however, indicate that fetal cholesterol homeostasis and the rate limiting enzyme of cholesterol synthesis is influenced by maternal glucocorticoid administration. PMID- 2556469 TI - Bioreactivity of intratracheally administered slate dust in rats: incorporation of 14C-acetate into lung lipids. AB - The effect of intratracheally instilled slate dust on the phospholipid profile, and 14C-acetate incorporation into the lipids of lung lavage, whole lung tissue and its subcellular fractions, has been studied in rats. The acellular fraction of lung lavage showed a decrease in the phospholipid content at 4 days and then an increase at 40 days of dust exposure, whereas the cellular fraction showed the reverse. The order of 14C-acetate incorporation into total lipids and individual phospholipids showed a parallel trend. The rate of incorporation with total lipids of lung tissue was higher at the two stages of dust exposure and a similar pattern prevailed in the subcellular fractions, i.e. mitochondrial, microsomal and cytosolic fraction. Acetate incorporation was highest in mitochondria, followed by the microsomes. An increase in the microsomal and mitochondrial cholesterol levels was also observed. There was no significant change in the solvent-extracted 14C-counts of whole plasma, trichloroacetic acid (TCA) precipitate and TCA supernatant of plasma. The results indicate that slate dust causes an enhanced synthesis of pulmonary surfactant and other lung lipids and, therefore, has an effect on the metabolism of type II alveolar epithelial cells. PMID- 2556471 TI - Risks to health care workers from occupational exposure to hepatitis B virus, human immunodeficiency virus, and cytomegalovirus. AB - Transmission of hepatitis B, HIV-1 and CMV by percutaneous, mucosal, or contact with intact skin is of concern to health care personnel. This article summarizes current information regarding the risks of transmission of these agents and emphasizes infection control measures. PMID- 2556472 TI - In vivo priming of helper T cells in the absence of B cell activation. AB - This paper describes an adjuvant-free immunization regimen that results in the priming of T cells but not B cells. B10.A mice were primed s.c. with syngeneic spleen cells that had been pulsed with the peptide 81-104 derived from pigeon cytochrome c. The T cell response was measured by using a sensitive limiting dilution assay that measures lymphokine production. The precursor frequency of Ag specific cells found in these mice was indistinguishable from the frequency found in mice primed in the footpads with 81-104 in CFA. A striking difference in antibody induction was found, however, when these two immunization regimens were compared. Mice primed with 81-104 in CFA developed significant serum antibody responses against the peptide, whereas mice primed with Ag-pulsed spleen cells produced no detectable anti-peptide antibodies. This lack of antibody did not result from detectable differences in the T cells that were primed: no differences were seen in IL-2 and IL-4 production or in the ability to provide help to B cells in vitro. In vitro stimulation with LPS suggested that the B cells were not primed by the Ag-pulsed spleen cells. The B cells were not tolerized, however, because boosting the mice with Ag in CFA resulted in the induction of an antibody response. The failure to induce an antibody response by priming with Ag-pulsed spleen cells was not caused by the site of immunization or the total amount of Ag used for priming. The critical variable may be the introduction of the Ag on the surface of an APC; in this form, B cell Ag recognition was apparently inefficient, whereas T cell Ag recognition was optimal. PMID- 2556474 TI - Evidence for the existence of two forms of membrane tumor necrosis factor: an integral protein and a molecule attached to its receptor. AB - Plasma membranes were isolated from thioglycolate-induced peritoneal mouse macrophages and tested directly in a 51Cr-release assay against WEHI 164 tumor cells. These membranes showed anti-TNF antibody inhibitable killing of the TNF sensitive tumor cell line, indicating that membrane-associated TNF is present on mouse macrophages. In order to elucidate whether membrane TNF is an integral protein or a molecule attached to a receptor, cells and plasma membranes were treated with low pH buffer. A partial reduction in TNF activity was observed which could be restored by incubation with exogenous TNF. In a Western blot analysis the integral membrane TNF could be identified as the 26-kDa molecule on activated mouse macrophages. These results indicate that both forms of membrane associated TNF exist on macrophages and are responsible for cell-mediated cytotoxicity against TNF-alpha-sensitive targets. PMID- 2556473 TI - IL-1 activates the Na+/H+ antiport in a murine T cell. AB - One of the early events following growth factor exposure is elevation of intracellular pH, a process mediated by the Na+/H+ antiport. We studied the effects of human rIL-1 alpha (HrIL-1 alpha) on intracellular pH (pHi) and calcium ([Ca2+]i) in a murine T cell line (MD10 cells), which proliferates in response to IL-1 alone. By using the intracellularly trapped fluorescent dyes (2(1),7(1)-bis 2-carboxyethyl)-5(and -6) carboxyfluorescein) and indo-1, we monitored immediate to early changes of pHi and [Ca2+]i in response to HrIL-1 alpha. Exposure to HrIL 1 alpha (120 pM) leads to an early, sustained intracellular alkalinization (delta pH = + 0.09 +/- 0.03) that plateaus within 20 min. Lower concentrations of the monokine (12 pM, 1.2 pM) have a positive but not statistically significant effect on pHi. These effects parallel the degree of MD10 IL-1R saturation predicted by the KD (49 pM) as assessed by 125I-HrIL-1 alpha binding by MD10 cells (Bmax = approximately 1300). Both the MD10 IL-1 receptor KD and the HrIL-1 alpha concentration required to induce early measurable alkaline pH shifts, however, exceed by three orders of magnitude the HrIL-1 alpha ED50 (50 fM) required for MD10 proliferation. The IL-1-induced rise in pHi is both sodium dependent and amiloride sensitive, indicative of activation of the Na+/H+ antiport. Additionally, PMA (100 nM) and IL-2 (2 nM) alkalinize MD10 cells, with the rise in pHi as a result of PMA exceeding the maximal IL-1 effect (delta pH = + 0.13 +/ 0.04). Furthermore, although PMA alkalinizes cells previously exposed to HrIL-1 alpha, the monokine does not alter the pHi of PMA-treated MD10 cells. Importantly, intracellular alkalinization induced by either HrIL-1 alpha or PMA is inhibited by staurosporine (1 mu iM). Finally, HrIL-1 alpha does not change MD10 [Ca2+]i, in either an acute or sustained fashion. These results indicate that IL-1 activates the Na+/H+ antiport in T cells by a mechanism that is unrelated to changes in [Ca2+]i but may involve protein kinase C activation. PMID- 2556475 TI - My 43, a monoclonal antibody that reacts with human myeloid cells inhibits monocyte IgA binding and triggers function. AB - A mAb My 43 of the IgM isotype was obtained from a fusion of spleen cells immunized against human monocytes. This mAb inhibited monocyte binding of both soluble FITC-labeled IgA and IgA-coated E, whereas it did not inhibit IgG binding. The Ag recognized by My 43 was induced on HL-60 cells in parallel with IgA binding ability by 1-25 dihydroxy-vitamin D3 treatment. Phagocytosis of IgA coated E by monocytes and 1-25 dihydroxyvitamin D3-treated HL-60 cells was inhibited by My 43. Furthermore, a heteroantibody of My 43 x F(ab)'2 anti-E promoted phagocytic uptake of E by monocytes. Production of superoxide anion by IFN-gamma treated U-937 cells was stimulated by My 43 but not by other IgM mAb recognizing myeloid cells. By these criteria My 43 recognized a molecule capable of triggering function. Moreover, its binding reactivity, ability to block binding of IgA and IgA-complexes, and its ability to induce activation of IgA receptor bearing myeloid cells, are consistent with the possibility that My 43 reacts with the IgA receptor on these cells. PMID- 2556476 TI - Adenosine receptors are expressed during differentiation of monocytes to macrophages in vitro. Implications for regulation of phagocytosis. AB - Ingestion by phagocytes is known to be markedly enhanced by physiologic signals such as cytokines and extracellular matrix proteins which may be found in inflammatory sites. Little investigation has been made of mechanisms that may depress this increased rate of phagocytosis during resolution of inflammation. We show that adenosine can act as an inhibitor of phagocytosis by macrophages derived from in vitro culture of human peripheral blood monocytes. Adenosine (Ado) is equally effective at inhibiting IgG Fc and complement-mediated phagocytosis. However, Ado has no effect on phagocytosis by freshly isolated monocytes. Inhibition by Ado begins after 2 days in culture and reaches a plateau by 5 days; these kinetics of induction of inhibition of phagocytosis parallel an increase in specific Ado binding to the macrophage plasma membrane. Ado binds to cultured monocytes with a Kd of 6 microM. This affinity and the observation that 2-chloroadenosine and 5'-N-ethylcarboxamidadenosine are the most potent inhibitors of phagocytosis suggest that the Ado receptors expressed during monocyte differentiation are of the A2 type. The inhibition of phagocytosis may be mediated by cAMP, a second messenger coupled to A2 receptors in several cell types. Thus, plasma membrane expression of A2 receptors dramatically increases during monocyte differentiation in vitro. These data show that a potentially physiologic mediator can have very different effects on the function of monocytes and macrophages. This suggests a mechanism whereby phagocytic function at inflammatory sites can be down-regulated if and only if signals for the recruitment of new phagocytes have subsided. PMID- 2556477 TI - Asparagine-linked glycosylation of cytochrome b558 large subunit varies in different human phagocytic cells. AB - Cytochrome b558, an essential component of the respiratory burst of phagocytic cells, is the terminal electron donor to molecular oxygen that results in the formation of superoxide anion (O2-.). It is an integral membrane heterodimer that in neutrophils consists of a 22-kDa small subunit and a highly glycosylated 91 kDa large subunit. Identical core proteins often differ in glycosylation in different cell types and with some membrane glycoproteins, the glycosylation state may markedly affect function. In the present study, antisera reactive with cytochrome b558 large subunit was used for immunoblot analysis of the glycosylation pattern of this subunit from different types of phagocytic cells. Striking variability in the apparent m.w. of this broadly banding subunit was detected in five different phagocytic cell types (neutrophils 78,000 to 93,000; eosinophils 74,000 to 115,000; monocytes 82,000 to 99,000; dibutyryl cyclic AMP induced HL-60 cells 79,000 to 103,000; dimethyl sulfoxide-induced HL-60 cells 77,000 to 110,000). However, after complete cleavage of N-linked oligosaccharides with endoglycosidase F, the core peptide of cytochrome b558 large subunit from these different cell types had the same Mr (58,000). Inhibition of N glycosylation with tunicamycin in differentiating HL-60 cells resulted in the synthesis of immunoreactive protein of the same m.w. and banding pattern as seen after endoglycosidase F cleavage. These tunicamycin treated cells retained some capacity to generate superoxide anion when stimulated with PMA. We conclude that the identity of the N-linked oligosaccharides of the cytochrome b558 large subunit differ in various phagocytic cells. All N-linked glycans on cytochrome b558 in all cell types examined were of the complex type as defined by resistance to endoglycosidase H cleavage. N-linked glycosylation of the cytochrome b558 large subunit may not be essential for activation of the respiratory burst. PMID- 2556479 TI - IL-2 protects T lymphocytes from glucocorticoid-induced DNA fragmentation and cell death. AB - In the IL-2-dependent T cell clone CTLL-2, dexamethasone, a synthetic glucocorticoid, induces a suicide program characterized by the early degradation of chromatin in oligonucleosome-length fragments which precedes the loss of cell viability by 2 to 4 h. These effects are most likely mediated through the interaction with a specific glucocorticoid receptor as suggested by the structure activity relationship of the various steroids tested. Incubation of nuclei of glucocorticoid-untreated cells in the presence of calcium and magnesium ions induces the cleavage of DNA in the linker region between nucleosomes, suggesting that fragmentation of chromatin in intact cells by glucocorticoids may involve the activation of a preexisting endonuclease. Interestingly, the presence of a saturating dose of IL-2 during the treatment of CTLL-2 cells with glucocorticoids completely blocks the cell death program. PMID- 2556478 TI - Regulation of Fc gamma receptor expression and phagocytosis of a human monoblast cell line U937. Participation of cAMP and protein kinase C in the effects of IFN gamma and phorbol ester. AB - We investigated the positive and negative effects of IFN-gamma, PMA, dibutyryl cAMP (Bt2cAMP), dexamethasone and transforming growth factor-beta (TGF-beta) on Fc gamma R subtype expression and phagocytosis of a human monoblast cell line, U937. IFN-gamma increased and Bt2cAMP decreased Fc gamma RI expression determined by a mAb 32.2, whereas PMA and Bt2cAMP increased Fc gamma RII expression determined by a mAb IV-3. Phagocytosis was measured microscopically by counting ingested aggregated human IgG- or BSA-treated ox E (Eo'-IgG or Eo'-BSA). IFN gamma increased the phagocytosis of Eo'-IgG but not that of Eo'-BSA, and PMA increased the phagocytosis of both Eo'-IgG and Eo'-BSA. Bt2cAMP decreased both basal and IFN-gamma- and PMA-augmented phagocytosis of U937 cells. Dexamethasone also inhibited both basal and IFN-gamma-augmented Fc gamma RI expression and PMA augmented Fc gamma RII expression and phagocytosis, but did not affect IFN-gamma augmented phagocytosis of Eo'-IgG. The augmentation of phagocytosis of Eo'-IgG by IFN-gamma thus seems to be due mainly to the increased internalizing process rather than to increased Fc gamma RI expression. TGF-beta slightly decreased Fc gamma R expression. In a study of the participation of protein kinase C (PK-C), it was found that H-7, a PK-C inhibitor, did not inhibit either IFN-gamma- or PMA enhanced Fc gamma RI and Fc gamma RII expression, respectively, and 1-oleoyl-2 acetylglycerol and N-(6-phenylhexyl)-5-chloro-1-naphthalenesulfonamide, both PK-C activators, did not show any apparent increase in Fc gamma R expression and phagocytosis. These results show that Fc gamma RI and Fc gamma RII expression on U937 cells is regulated by different mechanisms and that IFN-gamma and PMA play their roles in Fc gamma R expression and phagocytosis by different pathways. It is possible that cAMP but not PK-C plays an important role in the regulation of Fc gamma R expression and phagocytosis. PMID- 2556480 TI - Activation of the superoxide-forming NADPH oxidase of macrophages requires two cytosolic components--one of them is also present in certain nonphagocytic cells. AB - The superoxide-forming NADPH oxidase of resting macrophages can be activated in a cell-free system by certain anionic amphiphiles, most notably SDS. Activation requires the cooperation of membrane-associated and cytosolic components. We now report that at least two cytosolic factors are required for SDS-elicited activation of NADPH oxidase of guinea pig macrophages. Treatment of cytosol with ammonium sulfate at 37% saturation led to the partition of the two factors in the supernatant and precipitate fractions (termed components sigma 1 and sigma 2, respectively). Although each fraction by itself was inactive, recombining them resulted in complete recovery of the original ability of native cytosol to support SDS-elicited superoxide production by octyl-glucoside solubilized macrophage membranes. Both components are proteins, as shown by their susceptibility to trypsin and proteinase K, and were inactivated by heating at 60 degrees C. sigma 2, but not sigma 1, was inactivated by treatment with the covalent sulfhydryl reagent N-ethylmaleimide. On high-performance gel filtration, sigma 1 was found to have a molecular mass of 30 to 52 kDa, whereas sigma 2 eluted with molecules of 150 to 440 kDa. Component sigma 1 was partially purified from the ammonium sulfate supernatant fraction of cytosol by hydrophobic interaction chromatography followed by gel filtration. A material behaving like sigma 1 was also found to be present in the cytosol of guinea pig thymus cells, lymph node lymphocytes and brain and of the mouse myeloma cell line MOPC 315. However, sigma 2 appears to be strictly phagocyte specific. The molecular characteristics of sigma 1 components from nonphagocytic cells were similar to those of macrophage sigma 1, as shown by their presence in the supernatant, after treatment of cytosol with ammonium sulfate at 37% saturation, a molecular mass close to 30 to 52 kDa and a similar behavior on hydrophobic interaction chromatography. These findings raise the possibility that cytosolic component sigma 1 might be the bearer of a cellular function, more general than the one suggested by its role in the activation of NADPH oxidase of phagocytes. PMID- 2556482 TI - [Mesoblastic nephroma. Unusual renal tumor in adults. Apropos of a case]. AB - We report an observation concerning a girl 18 years old, who present a nephroma discovered following nephric colic and total and intermittent hematuria. The anatomo-pathological examination reveals a mesoblastic nephroma. This type of tumour with mild appearance is scarce in old child and exceptional in adult. In almost the whole of cases, it occurs in newborns and mursling from which designation of congenital mesoblastic nephroma is derived. Its good prognosis justifies its classification separately, out of nephroblastoma. PMID- 2556481 TI - Predominant utilization of V beta 8+ T cell receptor genes in the H-2Ld restricted cytotoxic T cell response against the immediate-early protein pp89 of the murine cytomegalovirus. AB - Cytotoxic T cell responses to the murine Cytomegalovirus (MCMV) were elicited in BALB/c mice (H-2d) by infectious virus. Eight days after infection, MCMV-primed local lymph node T cells were either depleted for T cells expressing a V beta 8+ TCR or separated into V beta 8+ and V beta 8- subpopulations by a cell sorter using the mAb F23.1. T cells were then expanded in vitro under limiting dilution conditions in the presence of IL-2 and in the absence of viral Ag to avoid selection by Ag in vitro. Frequencies of CTL precursors specific for the Immediate-Early-Ag 1 of MCMV and restricted to H-2Ld were determined. L cells of the endogenous haplotype H-2k cotransfected with the genes for MCMV-IE 1 and H 2Ld were used as target cells. Detection of a CTL response required previous priming of the animals by infection in vivo (less than 1/10(6) for nonimmunized animals). In primed animals CTL precursors of this specificity and restriction were three to fivefold more frequent in the V beta 8+ population (1/9.900 to 1/22.300) than in the V beta 8- population (1/57.000 to 1/87.200). Control experiments showed that frequencies were not influenced by the treatment with the anti-V beta 8-antibody and the fluorescein-labeled anti-Ig itself. V beta 8+ and V beta 8- T cells did not reveal any frequency differences when several other responses were determined (TNP-specific self-restricted CTL precursor; Th cells specific for keyhole limpet hemocyanin or Listeria monocytogenes). PMID- 2556483 TI - Cerebral blood flow response to the tissue temperature in tumour and brain tissues. AB - The response of regional-cerebral blood flow (rCBF) to change in the tissue temperature was studied using normal and tumour-bearing monkeys. The local brain was selectively heated by the external microwave irradiation, while the body was kept hypothermic (30.1 +/- 0.1 degrees C, mean +/- standard error) by immersion in a cold water bath. The rCBF in brain and/or tumour tissues was sequentially measured by inhalation hydrogen clearance method. In the normal animal study (n = 7), rCBF changed in response to the tissue temperatures over a range of 29.4-40.7 degrees C with a constant rate 15.2% per degree Celsius change. Similarly, rCBF in the tumour-bearing animals (n = 7) changed proportionately with change in the tissue temperatures over a range of 28.4-42.5 degrees C in tumour and 27.6-41.8 degrees C in brain tissue. The rate in rCBF change per degree Celsius was 6.5% for tumour, which was significantly smaller than that for brain tissue (13.5%) (P less than 0.01). These results indicated that rCBF can be controlled by the defined application of selective heating with temperatures ranging from shallow hypothermia to modest hyperthermia. Vascular response to temperatures in the tumour and brain tissues may play a significant role in the application of heat to brain tumour treatment. PMID- 2556484 TI - Low molecular weight heparin (Alfa LHWH) compared with unfractionated heparin in prevention of deep-vein thrombosis after hip fractures. AB - Efficacy and safety of a low molecular weight heparin (Alfa LMWH) was compared with unfractionated heparin (UFH) in the prevention of post-operative venous thromboembolism after hip fractures. Forty-nine patients were randomized to treatment with Alfa LMWH 7500 anti-Xa coagulometric units twice daily or with UFH 5000 IU t.i.d. Screening for thrombosis was performed with 125-I-fibrinogen leg scanning and strain-gauge plethysmography. Positive results were confirmed by venography. Five patients in the Alfa LMWH group (20 per cent) developed venographycally proven deep vein thrombosis (DVT) versus seven (29 per cent) in the UFH group. One pulmonary embolism and two deaths occurred in the UFH group and none in the LMWH group. No differences in haemorrhagic complications and blood loss indices were observed. Alfa LMWH appears to be a promising drug for prevention of venous thromboembolism after orthopaedic surgery. A "flexible" schedule of administration is proposed on the basis of the results of plasma anti Xa assays. PMID- 2556485 TI - [Result of registration and follow-up system of gestational trophoblastic disease in Shizuoka Prefecture (from 1977 to 1988)--recent trend and choriocarcinoma following term gestation]. AB - The remission rate of choriocarcinoma has greatly improved since the introduction of effective multiagent chemotherapy combined with aggressive surgical therapy and radiotherapy. In addition to these, the registration and follow-up of gestational trophoblastic disease (GTD) also has been playing an important role in the early detection and treatment of choriocarcinoma following hydatidiform mole. The system for the registration and follow-up of GTD was started in 1977 in Shizuoka Prefecture. In the present series, the results obtained with this registration and follow-up system from 1977 to 1988 in Shizuoka Prefecture were reviewed and analysed. 1) One thousand, nine hundred and twenty-five cases of hydatidiform mole, 68 cases of invasive mole, 70 cases of persistent trophoblastic disease and 48 cases of choriocarcinoma were registered in 12 years. The overall registration rate was 97.4%. 2) The number of cases of hydatidiform mole registered has decreased from about 180 cases to about 140 cases per year, probably due to the decreasing birth rate. 3) The number of cases of choriocarcinoma registered has recently been decreasing significantly and the number of cases of registered invasive mole and persistent trophoblastic disease has decreased slightly. 4) Antecedent pregnancy with choriocarcinoma including clinical choriocarcinoma has been changing from "post-molar" to "post-term" in the past 12 years. The prognosis of the patient with choriocarcinoma following hydatidiform mole has improved by the early detection and treatment since the introduction of the registration and follow-up system. More attention should be paid to choriocarcinoma following term gestation not yet included in the registration and follow-up system to facilitate early detection and treatment. PMID- 2556486 TI - [Characteristic ultrasonographic findings of the Krukenberg tumor]. AB - Researching into the possibility of ultrasonographic differential diagnosis between the Krukenberg tumor and primary ovarian cancer, we investigated ultrasonograms of 14 Krukenberg tumors, 14 primary ovarian cancers and 5 rare ovarian tumors. Specifically we examined in each the echo pattern of the tumor wall, solid part and cystic part. The results were as follows. 1) Thirteen of the 14 cases of Krukenberg tumor demonstrated a clear tumor margin, irregular hyperechoic solid pattern and moth-eaten cyst formation. These characteristic ultrasonographic features might reflect the specific pathological findings of the Krukenberg tumor. In only one case was there a clear tumor margin, large central cyst formation and diffuse thick solid part around the cysts. 2) The primary ovarian cancers had an indistinct tumor margin, irregular low echoic solid pattern, clear small cyst formation, papillary proliferation and irregular thickness of the septum. Taking the ultrasonographic characteristic findings of the Krukenberg tumors into consideration, eleven of 14 primary ovarian cancers (78.6%) could be distinguished ultrasonographically from the Krukenberg tumor. 3) In rare cases, only a carcinosarcoma was unable to be distinguished ultrasonographically from the Krukenberg tumor. that make it possible to distinguished it ultrasonographically from primary ovarian cancer. PMID- 2556488 TI - Sensitivity of in situ hybridization for detection of human papillomavirus (HPV) as studied in HPV type 16-transfected cells. PMID- 2556487 TI - Effect of naloxone on hormonal changes during exercise. AB - It is well established that prolactin release during exercise is one of the important factors in exercise-induced menstrual dysfunction. The purpose of this study is to clarify the mechanisms of prolactin release during exercise. Ten female athletes measured their BBT every morning. They performed incremental exercise on a cycle ergometer, with or without naloxone, on the 5th to 8th days of the follicular phase. Three minutes before the exercise, 0.4mg of naloxone was injected intravenously and a further 1.6mg/hr of naloxone was continuously infused during exercise. Blood samples were collected after 60 minutes bed rest (Rest), at the time when the heart rates reached 150 bpm (Submax), the point of exhaustion (Max) during exercise and after 60 minutes bed rest following exercise (After 1hr). The levels of prolactin in serum, dopamine, beta-endorphin. VIP and ACTH in the plasma were measured. Whereas prolactin increased significantly at Submax (p less than 0.05) and Max (p less than 0.001), the increase in prolactin was suppressed by the administration of naloxone (p less than 0.05). Dopamine showed no remarkable change during exercise, with or without naloxone. There were significant increases in beta-endorphin at Max (p less than 0.001), VIP at Submax and Max (p less than 0.001), but these increases were suppressed by the administration of naloxone (p less than 0.001). ACTH which had markedly increased at Submax (p less than 0.025) and Max (p less than 0.001) showed a slight tendency to decrease following the administration of naloxone, but there were no significant differences in both groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556489 TI - [A case of insulinoma with the splenic hamartoma presenting a vascular blush in arteriography]. PMID- 2556490 TI - Heterogeneity in cell recovery and superoxide production in buoyant, density defined subpopulations of human alveolar macrophages from healthy volunteers and sarcoidosis patients. AB - Reactive oxygen species (ROS) are ubiquitous compounds produced by phagocytes with important roles in both host defense and pulmonary inflammation. Enhanced ROS metabolism by alveolar macrophages (AM) has been previously demonstrated in various interstitial lung diseases including sarcoidosis. We studied 17 healthy, nonsmoking volunteers and 10 patients with sarcoidosis by bronchoalveolar lavage, and separated AM on discontinuous Percoll gradients to determine patterns of airspace cell recovery and the corresponding ROS metabolism of density-defined AM subpopulations. AM subpopulations were largely purified from contaminating granulocytes, thereby allowing more accurate estimation of ROS metabolism by AM. In bronchoalveolar lavage material from sarcoidosis patients, increased recovery of cells of high (1.075 gm/ml) density was found, which contrasted with the pattern seen in the volunteers in whom cells of lowest density (1.045 gm/ml) predominated. In addition, dense cells from sarcoidosis patients exhibited enhanced stimulated ROS metabolism compared with cells of similar density obtained from the volunteers or with sarcoid cells of lower density. At least three mechanisms may contribute to increased lung oxidative burden in sarcoidosis. The combination of increased bronchoalveolar lavage cell counts, increased cell recovery at high density, and increased cell function produced substantial increases in the total oxidative burden imposed on the lungs of sarcoidosis patients by airspace cells. We conclude that AM metabolism of ROS is dependent on the density and, by implication, the maturity of the cells and that regulation of AM ROS metabolism differs markedly between sarcoidosis patients and healthy volunteers. PMID- 2556491 TI - Influence of pertussis toxin on parathyroid hormone stimulated cyclic AMP production and phosphate transport in opossum kidney cells. AB - There is evidence that guanine nucleotide-sensitive (G) proteins intervene in the activation of adenylate cyclase by parathyroid hormone (PTH). Furthermore, recent studies suggest that G proteins may be involved in the activation by PTH of phospholipase C, with subsequent elevation of diacylglycerol, inositol trisphosphate, and intracellular calcium. Since G proteins may be involved in both transduction systems postulated to mediate the actions of PTH, the present studies were performed to evaluate the influence of pertussis toxin, which prevents receptor-mediated activation of G proteins, on the effects of PTH in opossum kidney (OK) cells. In OK cell membranes, pertussis toxin catalyzed the adenosine diphosphate (ADP) ribosylation of a protein with a molecular weight of 41 kd on SDS-PAGE. Cholera toxin catalyzed the ribosylation of two proteins of molecular weight 52 and 45 kd. Pretreatment of the cells with pertussis toxin abolished the labelling of this 41 kd protein, confirming the access of the toxin into the cells and the presence of pertussis toxin-sensitive substrates. The ribosylation of the cholera toxin substrates was unaffected by pertussis toxin pretreatment of the cells. Treatment of OK cells with pertussis toxin did not change the basal levels of cyclic AMP, but increased the levels of cyclic AMP in response to bPTH 1-34 from 355 +/- 17 to 449 +/- 20 pmoles cyclic AMP per 5 minutes per culture. These results were consistent with the inactivation of an inhibitory G protein. Furthermore, PTH-stimulated cyclic AMP generation was inhibited by norepinephrine from 362 +/- 10 to 228 +/- 18 pmole cyclic AMP per 5 minutes per culture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556493 TI - Congenital mesoblastic nephroma: report of a case. AB - Congenital mesoblastic nephroma (CMN) is a rare renal tumor with a distinct pathology and a unique clinical, therapeutic, and prognostic pattern. The characteristic pathological feature is uniform mesenchymal spindle-shaped cells arranged in interlacing bundles. The prognosis of typical CMN is believed to be excellent by nephrectomy alone. We report a typical case of congenital mesoblastic nephroma which was prenatally detected, and was associated with maternal polyhydramnios. PMID- 2556492 TI - Prevalence of antibodies to herpes simplex virus types 1 and 2 in pregnant women, and estimated rates of infection. AB - There has been a recent increase in notifications of genital herpes but it is not known whether this has been reflected in the pregnant population. We have therefore carried out a study to determine the prevalence of herpes simplex antibodies in pregnant women and to estimate the incidence of primary infection. Sera were collected from 3533 women at antenatal clinics and tested for total antibodies to herpes simples virus (HSV), and if positive, for specific antibodies to HSV-2. Estimates of HSV-1 seroprevalence were derived from the HSV 2 seronegative population. HSV-1 seroprevalence was nearly 100% in black women born in Africa or the Caribbean and 60-80% in white, Asian and UK born black women. It was lower in women in non-manual employment. HSV-2 seroprevalence was related to age, rising from 0 at age 16 to 40% at age 35 in black women, and to about 10% in Asian and white women. The estimated incidence of primary HSV-2 infection during pregnancy, per 1000 pregnancies, was about 2.4 in Asian women, 5 in white women, and 20 in black women. Estimates of the incidence of neonatal infection were derived from these figures and compared to the nationally reported rates. PMID- 2556494 TI - Influence of sodium-calcium exchange on calcium current rundown and the duration of calcium-dependent chloride currents in pituitary cells, studied with whole cell and perforated patch recording. AB - The whole cell patch-clamp technique, in both standard and perforated patch configurations, was used to study the influence of Na+-Ca++ exchange on rundown of voltage-gated Ca++ currents and on the duration of tail currents mediated by Ca++-dependent Cl- channels. Ca++ currents were studied in GH3 pituitary cells; Ca++-dependent Cl- currents were studied in AtT-20 pituitary cells. Na+-Ca++ exchange was inhibited by substitution of tetraethylammonium (TEA+) or tetramethylammonium (TMA+) for extracellular Na+. Control experiments demonstrated that substitution of TEA+ for Na+ did not produce its effects via a direct interaction with Ca++-dependent Cl- channels or via blockade of Na+-H+ exchange. When studied with standard whole cell methods, Ca++ and Ca++-dependent Cl- currents ran down within 5-20 min. Rundown was accelerated by inhibition of Na+-Ca++ exchange. In contrast, the amplitude of both Ca++ and Ca++-dependent Cl- currents remained stable for 30-150 min when the perforated patch method was used. Inhibition of Na+-Ca++ exchange within the first 30 min of perforated patch recording did not cause rundown. The rate of Ca++-dependent Cl- current deactivation also remained stable for up to 70 min in perforated patch experiments, which suggests that endogenous Ca++ buffering mechanisms remained stable. The duration of Ca++-dependent Cl- currents was positively correlated with the amount of Ca++ influx through voltage-gated Ca++ channels, and was prolonged by inhibition of Na+-Ca++ exchange. The influence of Na+-Ca++ exchange on Cl- currents was greater for larger currents, which were produced by greater influx of Ca++. Regardless of Ca++ influx, however, the prolongation of Cl- tail currents that resulted from inhibition of Na+-Ca++ exchange was modest. Tail currents were prolonged within tens to hundreds of milliseconds of switching from Na+- to TEA+-containing bath solutions. After inhibition of Na+-Ca++ exchange, tail current decay kinetics remained complex. These data strongly suggest that in the intact cell, Na+-Ca++ exchange plays a direct but nonexclusive role in limiting the duration of Ca++-dependent membrane currents. In addition, these studies suggest that the perforated patch technique is a useful method for studying the regulation of functionally relevant Ca++ transients near the cytoplasmic surface of the plasma membrane. PMID- 2556495 TI - Veratridine modification of the purified sodium channel alpha-polypeptide from eel electroplax. AB - In the interest of continuing structure-function studies, highly purified sodium channel preparations from the eel electroplax were incorporated into planar lipid bilayers in the presence of veratridine. This lipoglycoprotein originates from muscle-derived tissue and consists of a single polypeptide. In this study it is shown to have properties analogous to sodium channels from another muscle tissue (Garber, S. S., and C. Miller. 1987. Journal of General Physiology. 89:459-480), which have an additional protein subunit. However, significant qualitative and quantitative differences were noted. Comparison of veratridine-modified with batrachotoxin-modified eel sodium channels revealed common properties. Tetrodotoxin blocked the channels in a voltage-dependent manner indistinguishable from that found for batrachotoxin-modified channels. Veratridine-modified channels exhibited a range of single-channel conductance and subconductance states. The selectivity of the veratridine-modified sodium channels for sodium vs. potassium ranged from 6-8 in reversal potential measurements, while conductance ratios ranged from 12-15. This is similar to BTX-modified eel channels, though the latter show a predominant single-channel conductance twice as large. In contrast to batrachotoxin-modified channels, the fractional open times of these channels had a shallow voltage dependence which, however, was similar to that of the slow interaction between veratridine and sodium channels in voltage-clamped biological membranes. Implications for sodium channel structure are discussed. PMID- 2556496 TI - Voltage-dependent inactivation of slow calcium channels in intact twitch muscle fibers of the frog. AB - Inactivation of slow Ca2+ channels was studied in intact twitch skeletal muscle fibers of the frog by using the three-microelectrode voltage-clamp technique. Hypertonic sucrose solutions were used to abolish contraction. The rate constant of decay of the slow Ca2+ current (ICa) remained practically unchanged when the recording solution containing 10 mM Ca2+ was replaced by a Ca2+-buffered solution (126 mM Ca-maleate). The rate constant of decay of ICa monotonically increased with depolarization although the corresponding time integral of ICa followed a bell-shaped function. The replacement of Ca2+ by Ba2+ did not result in a slowing of the rate of decay of the inward current nor did it reduce the degree of steady state inactivation. The voltage dependence of the steady-state inactivation curve was steeper in the presence of Ba2+. In two-pulse experiments with large conditioning depolarizations ICa inactivation remained unchanged although Ca2+ influx during the prepulse greatly decreased. Dantrolene (12 microM) increased mechanical threshold at all pulse durations tested, the effect being more prominent for short pulses. Dantrolene did not significantly modify ICa decay and the voltage dependence of inactivation. These results indicate that in intact muscle fibers Ca2+ channels inactivate in a voltage-dependent manner through a mechanism that does not require Ca2+ entry into the cell. PMID- 2556497 TI - Decay of the slow calcium current in twitch muscle fibers of the frog is influenced by intracellular EGTA. AB - The mechanism(s) of the decay of slow calcium current (ICa) in cut twitch skeletal muscle fibers of the frog were studied in voltage-clamp experiments using the double vaseline-gap technique. ICa decay followed a single exponential in 10 mM external Ca2+ and 20 mM internal EGTA solutions in all pulse protocols tested: single depolarizing pulses (activation protocol), two pulses (inactivation protocol), and during a long pulse preceded by a short prepulse (400 ms) to 80 mV (tail protocol). In single pulses the rate constant of ICa decay was approximately 0.75 s-1 at 0 mV and became faster with larger depolarizations. ICa had different amplitudes during the second pulses of the inactivation protocol (0 mV) and of the tail protocol (-20 to 40 mV) and had similar time constants of decay. The time constant of decay did not change significantly at each potential after replacing 10 mM Ca2+ with a Ca2+-buffered solution with malate. With 70 mM intracellular EGTA and 10 mM external Ca2+ solutions, ICa also decayed with a single-exponential curve, but it was about four times faster (approximately 3.5 s-1 at 0 mV pulse). In these solutions the rate constant showed a direct relationship with ICa amplitude at different potentials. With 70 mM EGTA, replacing the external 10 mM Ca2+ solution with the Ca2+-buffered solution caused the decay of ICa to become slower and to have the same relationship with membrane potential and ICa amplitude as in fibers with 20 mM EGTA internal solution. The mechanism of ICa decay depends on the intracellular EGTA concentration: (a) internal EGTA (both 20 and 70 mM) significantly reduces the voltage dependence of the inactivation process and (b) 70 mM EGTA dramatically increases the rate of tubular calcium depletion during the flow of ICa. PMID- 2556498 TI - Diffuse Lewy body disease: correlative neuropathology using anti-ubiquitin immunocytochemistry. AB - Diffuse Lewy body disease is an important pathological substrate of the common syndrome of parkinsonian dementia. The new technique of anti-ubiquitin immunocytochemistry has been used in a correlative quantitative neuropathological study of fifteen cases of diffuse Lewy body disease, showing that the severity of dementia is related to cortical Lewy body density, whilst subcortical abnormalities make a much less significant contribution. Cortical senile plaques also appear to be part of the pathology of diffuse Lewy body disease and should not therefore be used as an isolated diagnostic criterion for Alzheimer's disease. Diagnostic criteria for diffuse Lewy body disease are discussed. PMID- 2556500 TI - Sensory neuropathy as the initial manifestation of primary biliary cirrhosis. PMID- 2556501 TI - Rapid recovery after delayed myelopathy from electrical burns. PMID- 2556499 TI - Resistance to ischaemic conduction failure in chronic hypoxaemia and diabetes. AB - Median nerve function was studied in twelve diabetic subjects, six subjects with chronic hypoxaemia and ten control subjects. Resistance to ischaemic conduction failure (RICF), a characteristic electrophysiological feature of diabetic neuropathy, was assessed by measuring the decline in median nerve action potential amplitude at minute intervals for up to 20 minutes while the arm was rendered ischaemic. Initial nerve conduction velocity and action potential amplitude was similar in all three groups. Following the onset of ischaemia the time to a 50% reduction in action potential amplitude was prolonged in both diabetic subjects and hypoxaemic subjects compared with controls. After 20 minutes of ischaemia no control subject had persisting nerve function, while function remained in 5 (80%) of hypoxaemic subjects and 10 (83%) of diabetic subjects. The time to a 50% reduction in action potential amplitude during ischaemia correlated with the blood oxygen saturation among the hypoxic subjects and haemoglobin Alc among diabetic subjects. These results are consistent with the hypothesis that hypoxia has a role in the pathogenesis of resistance to ischaemic conduction failure in diabetes. PMID- 2556502 TI - Coxsackie B5 papillitis. PMID- 2556503 TI - Alzheimer's disease: amyloid plaques in the cerebellum. AB - Two specific silver-staining methods demonstrating either extracellular amyloid and/or precursors of amyloid or intraneuronal neurofibrillary changes were used to examine cerebellar pathology in cases of presenile and senile dementia of the Alzheimer type, cases of Down's syndrome, and non-demented controls. The sensitivity of the techniques permitted visualization of large numbers of amyloid deposits in the cerebellar cortex of demented individuals. Similarly large numbers of amyloid deposits were not found in the cerebella of non-demented individuals. Neurofibrillary changes were absent. The majority of amyloid plaques occurred in the molecular layer. Quite a number of these displayed large diameters extending from the upper surface down to the Purkinje cell layer. Within the granular layer and white matter the plaques were less frequently encountered and they were less voluminous than those of the molecular layer. The cerebellar amyloid plaques were morphologically different and could easily be distinguished from the cerebellar plaques found in transmissible spongiform encephalopathies. PMID- 2556504 TI - Immunocytological and histochemical correlation in Kearns-Sayre syndrome with mtDNA deletion and partial cytochrome c oxidase deficiency in skeletal muscle. AB - We report histochemical, immunocytochemical, biochemical and molecular studies of skeletal muscle from a 23-year-old man with Kearns-Sayre syndrome. Southern blot analysis revealed a 4.7 kb heteroplasmic deletion of the mitochondrial DNA mapping within genes coding for subunits of complexes I, IV and V of the respiratory chain and for tRNA. Cytochrome c oxidase activity was decreased by 30% in isolated muscle mitochondria, without alteration of the Km. Histochemical and immunocytochemical correlation studies for cytochrome c oxidase revealed a lack of activity in 34% of individual muscle fibers including all the typical ragged-red fibers and a low percentage of immunodeficient fibers. PMID- 2556506 TI - Cross-communication between acetylcholine and VIP in controlling catecholamine secretion by affecting cAMP, inositol triphosphate, protein kinase C, and calcium in rat adrenal medulla. AB - The purpose of the present study was to determine the molecular mechanism of stimulatory actions of ACh and vasoactive intestinal polypeptide (VIP) by determining the role of various second messengers in the neurohumoral secretion. Toward such a goal, we measured cAMP, cGMP, protein kinase (PKC) activity, 3H inositol triphosphate (3H-IP3), and 45Ca uptake in the adrenal medulla subjected to various treatments. Stimulation of splanchnic nerve endings increased 45Ca uptake, cAMP content, 3H-IP3, and PKC activity in the adrenal medulla. If muscarinic receptors of chromaffin cells were selectively activated by perfusion with muscarine, 3H-IP3 content and PKC activity were enhanced. Nicotine, on the other hand, increased only 45Ca uptake without affecting any other second messenger. Perfusion with VIP increased PKC activity and cAMP and 3H-IP3 content. None of the procedures affected cGMP content. Interplay among various second messengers was further investigated by studying interactions of nicotinic, muscarinic, and VIP-ergic receptors in modulation of catecholamine (CA) secretion and by using agents known to activate specific second messengers (e.g., forskolin, phorbol esters). Our results show that muscarine, VIP, and phorbol ester facilitated nicotine-evoked secretion by increasing PKC activity, and it was associated with an additional increase in 45Ca accumulation. On the other hand, secretion evoked by nicotine as well as muscarine was facilitated by forskolin without additional increase in 45Ca accumulation. A novel feature of the study is that ACh and VIP activate three types of receptors on chromaffin cells to stimulate and mutually facilitate the secretion of CA by generating various second messengers.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556505 TI - Hypothalamic expression of a novel gene product, VGF: immunocytochemical analysis. AB - VGF is the designation for a new 712 amino acid protein, regulated by nerve growth factor (NGF) in PC12 cells, that has not been previously described in the CNS. Northern blot analysis with a nick-translated VGF cDNA probe revealed a single band of mRNA in the brain with a molecular weight identical to that found in PC12 cells. The current paper presents a series of immunocytochemical studies of VGF expression with a focus on the hypothalamus. Two different antisera were raised against nonoverlapping amino acid sequences of a bacterial-expressed protein from the VGF gene cloned from PC12 cells. VGF immunoreactivity is strongly expressed in the rat suprachiasmatic nucleus (SCN), particularly in the dorsomedial part of the nucleus. The administration of colchicine to block axonal transport facilitates detection of the VGF immunoreactivity also in the ventrolateral suprachiasmatic nucleus. This protein appears to be the first one of limited neuronal distribution which is found in both dorsomedial SCN and ventrolateral SCN. Immunostaining of serial 1 micron SCN sections reveals co localization of VGF in cells which also contain vasopressin or vasoactive intestinal polypeptide. Weaker immunoreactivity is also found in the magnocellular paraventricular and supraoptic nuclei, where the VGF immunoreactivity co-localizes with oxytocin or vasopressin. Mutant Brattleboro rats which do not express vasopressin showed strong VGF immunoreactivity both in the dorsomedial SCN and in cells of the magnocellular neuronal systems, including cells which normally express vasopressin. When axonal transport of the protein is blocked by colchicine, VGF-immunoreactive cells in the hypothalamic arcuate, parvocellular paraventricular, and tuberomammillary nuclei can also be detected, in addition to weakly immunoreactive scattered cells in the hippocampus, amygdala, thalamus, and cortex. VGF immunoreactivity is strong in the axonal projections of SCN and weak in the axons of the paraventricular and supraoptic nuclei. With ultrastructural studies, VGF immunoreactivity is found in presynaptic boutons in the SCN and in axons in the neurohypophysis. Weak axonal staining is present in some regions of the hypothalamus and in the external and internal zones of the median eminence. Immunoreactivity is absent from the intermediate lobe of the hypophysis. In neonatal rats strong VGF immunoreactivity is found throughout the SCN at postnatal day 4 but not in the adjacent hypothalamus. VGF immunoreactivity is also seen in other areas of the brain in neonatal rats, including the lateral geniculate nucleus; while the staining in the dorsal lateral geniculate disappears in the adult, that in the intergeniculate leaflet, a visual center which projects to the SCN, remains.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2556507 TI - Identification of morphine- and cyclic AMP-regulated phosphoproteins (MARPPs) in the locus coeruleus and other regions of rat brain: regulation by acute and chronic morphine. AB - Regulation of cAMP-dependent protein phosphorylation by acute and chronic morphine was studied in the rat locus coeruleus (LC), a brain region that appears to play an important role in mediating morphine action in animals, including humans. Rats were treated chronically with morphine under conditions known to induce states of tolerance and dependence. Protein phosphorylation was then studied in extracts of LC and other brain regions with back-phosphorylation assays under different conditions and with 1- and 2-dimensional electrophoretic procedures. Evidence was obtained to suggest that chronic morphine increased the phosphorylation state of proteins of 165, 55, and 14-20 kDa and increased the total amount of proteins of 145, 71, 62, 58, and 51 kDa in the LC. Concomitant treatment of rats with naltrexone, an opiate receptor antagonist, blocked the ability of morphine to influence each of these proteins, indicating that morphine regulation of protein phosphorylation occurred through the specific activation of opiate receptors. Regulation of 165, 71, 62, 58, 55, and 51 kDa by chronic morphine was specific to the LC among the brain regions studied, whereas regulation of 145 and 14-20 kDa was also observed in the frontal cortex, neostriatum, and dorsal raphe. Most of the phosphoproteins whose phosphorylation state or total amount was increased by chronic morphine treatment in the LC in vivo were also shown to have their phosphorylation state decreased by acute morphine treatment in isolated LC nuclei ex vivo. In addition, the phosphorylation state of most of these morphine-regulated phosphoproteins was stimulated by forskolin or cAMP analogs in isolated LC and by cAMP in broken cell preparations of this brain region, supporting the view that these proteins are physiological substrates for cAMP-dependent protein kinase in the LC. Phosphoproteins regulated by morphine and cAMP were designated "MARPPs," morphine and cAMP-regulated phosphoproteins, whereas those regulated by morphine but not by cAMP were designated "MRPPs." Taken together, the results of this study indicate that chronic morphine produces specific alterations in cAMP-dependent protein phosphorylation in the LC and raise the possibility that regulation of these specific phosphoproteins contributes to the development of tolerance and/or dependence in these neurons. PMID- 2556508 TI - Multiple electrosensory maps in the medulla of weakly electric gymnotiform fish. II. Anatomical differences. AB - Both wave- and pulse-type species of weakly electric gymnotiform fish have 3 topographic maps of electroreceptive information in the electrosensory lateral line lobe (ELL). These maps receive identical input from trifurcating axons of phase- and amplitude-coding primary afferents (Carr et al., 1982; Heiligenberg and Dye, 1982). Physiological experiments in the ELL of the wave-type fish Eigenmannia show that the amplitude-coding pyramidal cells differ among maps with respect to receptive field size, sensitivity, rate of adaptation, and temporal frequency response (Shumway, 1989). This study investigated morphological correlates of the physiological differences among maps. Estimates of primary afferent convergence in Eigenmannia, based on map size, cell counts, and areas of terminal fields from intracellularly filled P-type primary afferents, suggest a 2 fold increase in convergence in the lateral map relative to the centromedial map. Similar differences in convergence between maps are found in the wave-type species Apteronotus leptorhynchus and the pulse-type fish Hypopomus occidentalis. The lateral and centrolateral maps in Hypopomus, however, show an even greater difference in convergence. Comparison of the efferent projections of pyramidal cells among the different maps of Eigenmannia indicates that cells from the 3 maps terminate in the same laminae of the torus semicircularis, but the maps differ in the strength of projection to particular laminae. In both wave-type species, the abundance of a class of interneurons which receives descending input and inhibits pyramidal cells (interneurons of the ventral molecular layer) differs among maps; the centromedial map has 10 times fewer neurons of this type than the other 2 maps. Cytochrome oxidase studies in all 3 species demonstrated increased levels of activity in the lateral map, within the region receiving descending input from the cerebellum. These results suggest that the primary anatomical bases of the physiological differences among maps are differences in the amount of primary afferent convergence, coupled with differences in descending input. PMID- 2556509 TI - Protection of murine foot tissue and transplantable tumor against Photofrin-II mediated photodynamic sensitization with WR-2721. AB - Four thiol-containing compounds, WR-2721, WR-149024, WR-168643 and WR-361, were compared as photoprotectors of murine feet. The protector doses were the maximal tolerated intraperitoneal doses, administered 24 h after injection of Photofrin II and 15 min before illumination with 630-nm laser light. While all four compounds were effective, only WR-2721 demonstrated a statistically significant attenuation of phototoxicity. WR-2721 was found to protect SMT-F tumors in the same mouse strain, using tumor growth delay and short-term control as endpoints. A comparison of the dose modification factors for foot and tumor responses indicated no therapeutic advantage in using WR-2721 during photodynamic treatment of these two tissues. PMID- 2556510 TI - Primary extrarenal Wilms' tumor in children. AB - We report three additional cases of primary extrarenal Wilms' tumor and review those cases previously documented. Analysis of the location, histopathology, treatment, and survival of these cases supports the following conclusions: Wilms' tumor may occur in an extrarenal location without primary renal involvement and must be included in the differential diagnosis of abdominal, pelvic, and inguinal masses; an extrarenal location supports a more frequent occurrence of ectopic metanephric blastema than was previously recognized or origin of Wilms' tumor from a more primitive mesodermal tissue; and the natural history and prognosis of extrarenal and renal Wilms' tumors appears similar. PMID- 2556511 TI - Neutrophil function, genotype and periodontal bone loss in the mouse. PMID- 2556512 TI - Pediatric ambulatory surgery: preparing the patient and parent for discharge. PMID- 2556513 TI - Advertising unleashed. PMID- 2556514 TI - Community hospitals--time to come off the fence. PMID- 2556515 TI - McConaghey memorial lecture 1988. The emergence of the discipline of general practice, its literature, ant the contribution of the College Journal. PMID- 2556516 TI - HIV infection and Scottish general practice: workload and current practice. AB - To estimate the effect of human immunodeficiency virus (HIV) infection on general practice, a postal survey was undertaken of one in three of all principals in Scotland. Of the 834 general practitioners who responded (78% response rate), 31% were working in practices with patients known to be infected with HIV. The estimated prevalence of known HIV infection in general practice was 19 per 100,000 population, and the estimated annual consultation rate for HIV related problems (including consultations by the 'worried well') was seven per 1000 population. Both statistics showed considerable variation between health boards, with peaks in Lothian and Tayside. Few practices had drawn up policies relevant to HIV infection, and the use of procedures for controlling infection was variable. Policies about HIV and for infection control tended to be more common in areas where the prevalence of HIV infection was higher. Most respondents were offering both opportunistic health education and counselling about HIV infection, especially to patients at high risk. Although general practitioners are responding positively to the increasing demands of HIV infection, there is an urgent need for policies, both national and local, to guide specific aspects of practice. PMID- 2556517 TI - Nocturnal asthma: a study in general practice. AB - Symptoms of nocturnal asthma were studied using questionnaires returned by 1199 general practitioners throughout the United Kingdom. Of 7729 asthmatic patients seen consecutively and prescribed a bronchodilator aerosol, 73% woke with asthma at least once a week and 39% woke nightly. The percentage of asthmatics waking at night at least once a week in this population, where 48% were prescribed corticosteroid aerosols, was very similar to the 74% found to have asthma attacks at night in an earlier study of new hospital referrals at a time when such medication was not available. While sampling bias cannot be excluded, the clinical characteristics and profile of medications found in this study are similar to other reports and the evidence suggests that the general practitioners were managing these patients carefully. There was an overall association between the patients' perception of the severity of their asthma and frequency of waking at night (P less than 0.001). However, 26% of 2928 patients waking every night regarded their asthma as mild. These patients were taking significantly less medication than those also waking nightly but assessing their asthma as severe (P less than 0.001). The seriousness of nocturnal symptoms may be underestimated by asthmatics and they should be asked specifically about the frequency of nocturnal waking. Those with nocturnal asthma had a generally higher frequency of allergic and non-allergic provoking factors, but no single factor distinguished these patients from those without nocturnal symptoms. There was a strong correlation between the frequency of nightly waking and the number of medications used (P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556518 TI - Randomized controlled trial of an educational booklet for patients presenting with back pain in general practice. AB - A randomized controlled trial was used to evaluate an educational booklet on back pain for patients presenting to five group practices during one calendar year. The booklet had no immediate effect on consultations for back pain, but in the period from two weeks to one year after presentation significantly fewer patients in the group receiving the booklet consulted with back pain (35.6%) than in the control group (42.2%) (P less than 0.05). There were no significant differences between the booklet and control groups in certified absence from work owing to back pain. Referral to hospital, referral to physiotherapy, admissions to hospital and laminectomies were all less common in the booklet group. The reduction in the combined referral rate to physiotherapy and hospital, and the reduction in laminectomy rate almost reached statistical significance at the 5% level. In replying to a questionnaire sent one year after entry to the study 94.1% of respondents in the booklet group said that they had read the book, 84.0% said that they found it useful, and 68.0% said that they still had a copy. Scores on a 15-item test of knowledge about back pain were significantly higher in the group of patients who had received the booklet than in the control group. The results suggest that the booklet had some effect in altering both the knowledge and behaviour of patients with back pain. The provision of an educational booklet was a method of giving information which was appreciated by both patients and doctors. PMID- 2556519 TI - Teenagers' health concerns: implications for primary health care professionals. AB - Four hundred and eighty five students, aged 13-15 years, at nine comprehensive schools in the London borough of Brent completed a questionnaire about health related behaviours and health concerns. Among general health concerns, most prominent were weight, acne, nutrition and exercise. There appeared to be a considerable unmet need to discuss sexual development, sexually transmitted diseases and contraception. A substantial proportion (16% of the girls and 3% of the boys) reported sexual abuse, but few wished to discuss this with a doctor or nurse. Although a relatively high percentage of the students smoked and a smaller percentage used alcohol or drugs regularly, there was little concern or interest in discussing these matters with a health professional. Most of the schools did not have a formal health education programme, and in none of them were health professionals available for discussion of the issues under study. There appears to be a need for more comprehensive health education in schools and for primary health care professionals, particularly general practitioners to raise these issues opportunistically with their teenage patients. PMID- 2556520 TI - Survey of practice annual reports. AB - A questionnaire was sent to those practices known by the Royal College of General Practitioners to have produced annual reports. Practices were asked what data they included, to whom the report was circulated, what problems and benefits they encountered, and their opinion of the government's proposal to encourage such reports. The data most frequently included in reports related to practice workload, and the main benefit which respondents mentioned was facilitation of planning within the practice. Few practices used data derived from outside the practice and several commented on the difficulty of obtaining good quality data from the practice. Although annual reports are useful as internal documents, standardization of definitions and format, with support from family practitioner committees and health authorities is essential if more practices are to be encouraged to produce such documents and if the information obtained from them is to have a broader role in planning for primary health care. PMID- 2556521 TI - Indigestion or infection? Unusual presentations of malaria in general practice. AB - Two cases of vivax malaria which presented atypically as abdominal pain are described. They highlight the need to consider malaria in the differential diagnosis of any acute febrile illness in a patient returning from an endemic malarial area. PMID- 2556523 TI - Preferences for salbutamol delivery systems among young asthmatics. PMID- 2556522 TI - Asthma--still a challenge for general practice. AB - Asthma is probably the commonest chronic disease in the United Kingdom, and its attendant morbidity extends outside the possible scope of the hospital sector. Innovations to improve the care of asthma must therefore come from general practice. The need for such care initiatives is demonstrated by the rising mortality and morbidity from this condition, and by the evidence of less than optimum treatment at both the primary and secondary care levels. This paper reviews this evidence, and considers possible solutions to the problems raised. Pragmatic guidelines are offered for the promotion of good asthma care, while the need for proper evaluative research is stressed. PMID- 2556524 TI - Circular random case analysis. PMID- 2556525 TI - Dangers of steam inhalations: case report. PMID- 2556526 TI - Elevation of serum cholesterol and triglycerides as a complication of amiodarone therapy. PMID- 2556528 TI - Computer prescribing. PMID- 2556527 TI - Referral of women with chronic pelvic pain. PMID- 2556529 TI - International travel medicine. PMID- 2556530 TI - What influences doctor's prescribing. PMID- 2556531 TI - Telephone advice in patient management. PMID- 2556532 TI - 'Brittle' diabetes. PMID- 2556533 TI - Night calls--the patients' view. PMID- 2556534 TI - Staff for general practice. PMID- 2556536 TI - Working for patients. PMID- 2556535 TI - Quality of care. PMID- 2556537 TI - What kind of college? PMID- 2556538 TI - Satisfaction with the NHS: what is it and can we measure it? PMID- 2556539 TI - Prescribing research: PACT to the future. PMID- 2556540 TI - Randomized controlled trial of anti-smoking advice by nurses in general practice. AB - Practice nurses are playing an increasingly prominent role in preventive care, including the provision of anti-smoking advice during routine health checks. A randomized controlled trial was designed to assess the effectiveness of anti smoking advice provided by nurses in helping smokers to stop smoking. A total of 14,830 patients aged 16-65 years from 11 general practices completed a brief questionnaire on general health, including smoking status, at surgery attendance. The doctor identified 4330 smokers and randomly allocated 4210 to control or intervention groups. The doctor asked those in the intervention group to make an appointment with the practice nurse for a health check. The attendance rate at the health check was 26%. Smokers were sent follow-up questionnaires at one month and one year, and those who did not respond to two reminders were assumed to have continued to smoke. There was no significant difference in reported cessation between the intervention and control groups at one month or one year. However, there was a significant difference in the proportion of patients who reported giving up within one month and who had not lapsed by one year--0.9% in controls and 3.6% in the intervention group (P less than 0.01). Nevertheless, the effect of the nurse intervention itself may be small as the sustained cessation rate in attenders was only 42.4% higher than in non-attenders. The deception rate in reporting cessation, as measured by urinary cotinine, was of the order of 25%. PMID- 2556541 TI - Diagnosis and management of female urinary incontinence in general practice. AB - In response to an invitation sent to women who had complained previously of regular incontinence, 65 women with regular incontinence were seen by their general practitioner. A diagnosis was made using a personally administered questionnaire and appropriate examination. Patients were placed in one of three diagnostic/management categories--stress, urge or stress/urge incontinence--and were given an appropriate treatment programme. Fifty six women were recruited as matched controls from non-responders while attending the surgery for other reasons. They underwent identical entry procedures but were not offered a treatment programme. All the patients were reassessed after 12 weeks at which time significant improvement in incontinence was reported by the treated women in the stress and urge categories compared with the controls. There was no significant difference in reported efficacy of treatment between age groups and treatment was shown to be effective irrespective of the duration of incontinence. This study shows that for the majority of women reporting incontinence the condition can be diagnosed by a general practitioner and significantly improved by appropriate intervention. PMID- 2556542 TI - London general practitioners' involvement with HIV infection. AB - A total of 2510 general practitioners in the London postal districts were circulated with a questionnaire about their involvement with patients with human immunodeficiency virus (HIV) infection--1261 (50%) replies could be analysed. Over half of the respondents had at least one patients who was HIV seropositive and most felt confident to handle such patients' psychosocial problems. Although almost 60% of doctors had attended at least one postgraduate teaching session on the acquired immune deficiency syndrome (AIDS), this had not allayed the anxieties of those doctors who were apprehensive about working with patients infected with HIV. Doctors had a positive attitude towards HIV infected patients and homosexuals, but wanted to be better informed by their patients or the hospital services. They seemed less prepared to deal with drug abusers. Doctors who had HIV positive patients and had graduated in the UK within the past 10 years were likely to have the greatest knowledge of, and most positive attitudes towards AIDS. PMID- 2556543 TI - HIV infection and Scottish general practice: knowledge and attitudes. AB - To assess the knowledge and attitudes of general practitioners about HIV infection, a postal survey was undertaken of one in three of all principals in Scotland; 834 (77.6%) responded. Respondents' knowledge about HIV was often limited, and they found the discussion of sexual behaviour difficult. Most were in favour of routine HIV testing being offered to patients, but against testing without consent. Most general practitioners considered consent unnecessary for the passing of information about HIV status between medical colleagues, but necessary for informing others, in particular the patient's family and sexual partners. Most general practitioners would accept high-risk and HIV-positive patients onto their lists but less than half would accept intravenous drug users. Most respondents did not feel at significant personal risk of HIV infection, but expressed reservations about many other aspects of HIV infection in general practice. If practitioners are to fulfil their potential for tackling the problems of HIV infection, they need increased resources and a policy for education and training that is responsive to local needs. PMID- 2556544 TI - Full-time women general practitioners--an invaluable asset. AB - It has been said that women doctors do not fulfil the same role as men doctors in general practice. This is inaccurate as previous studies of workload in general practice have not excluded from the analyses women principals who are part time, that is, have a less than full profit share at parity. In a postal questionnaire to 501 women principals 308 (62%) replied, of whom 143 (46%) were full time with respect to profit share. Respondents were asked to record aspects of workload over a four-week period for themselves and three of their full-time men partners. Analysis of their responses showed that they did an equal workload to their full time men partners in terms of number of surgeries, length of surgeries and number of home visits. Equal numbers of full-time women did out-of-hours work as their men partners and the number of nights and weekends on-call as well as use of deputies were similar. The analysis also showed that full-time women did more specialized clinics than men, thus emphasizing the special role of women doctors in preventive care. PMID- 2556545 TI - Treatment of hay fever. AB - The range of treatments for hay fever available to the general practitioner has changed considerably in recent years. New antihistamines have addressed the problem of sedation and moved towards one daily dose; nasally applied corticosteroids avoid the need for systemic steroid therapy and its potential adverse effect; and regulatory decisions have set a trend away from immunotherapy in general practice. However, knowledge about the mechanism of action of immunotherapy is increasing and new developments with improved safety profiles include allergen polymers, allergoids, oral immunotherapy and nasal immunotherapy. Choice of treatment depends, as always, on the individual circumstances of the patient and his or her disease. PMID- 2556546 TI - Practice activity analysis: collaboration between general practitioners and a family practitioner committee. AB - The Brent and Harrow family practitioner committee has supported a scheme to enable general practitioners to collect data relating to their practice activity. This paper reports on the operation of the scheme involving 76 general practitioners from 22 practices and the findings. Practice nurse activity was also included. The family practitioner committee undertook an analysis of the data and provided each participating general practitioner with a print-out of his or her consultation rates over a range of activity, enabling general practitioners to make comparisons both with their partners and with the averages for all the practices participating in the scheme. The essential aim is to provide structured information which enables general practitioners to look more objectively at their activity. The family practitioner committee gave an assurance that the figures would not be used to criticize individuals. PMID- 2556547 TI - Compulsory admission to hospital of an alcoholic patient under the mental health act. PMID- 2556548 TI - Blood sample transportation and the erythrocyte sedimentation rate. PMID- 2556549 TI - Surveillance of iron deficiency, anaemia and hypercholesterolaemia in rural pre school children. PMID- 2556550 TI - Dispensing costs. PMID- 2556551 TI - Can general practitioners counsel? PMID- 2556552 TI - Children with special educational needs. PMID- 2556554 TI - Stationery for medical records: 1912-21. PMID- 2556553 TI - Secrecy and the College. PMID- 2556555 TI - Quality assessment or quality control? PMID- 2556556 TI - Is there a future for general practice postgraduate education? ? PMID- 2556557 TI - William Pickles lecture 1989. Seeing sunflowers. PMID- 2556558 TI - Comparison of the workload of a trainer and trainee. AB - A general practitioner trainee's workload over one year was compared with a sample of his trainer's workload in order to identify and explain any differences. In line with previous studies the trainee saw relatively fewer patients with chronic diseases, fewer women of child-bearing age and fewer elderly patients but more children, more men and more acute problems. However, the trainee saw a marked excess of ear, nose and throat conditions and asthma which is explained by his special interest in these two subjects. Over the first six months the number of doctor-initiated consultations seen by the trainee increased steadily and remained fairly constant over the second six months. Periodic study throughout the year enabled the provision of extra experience in areas where the trainee had seen few cases. PMID- 2556559 TI - Diabetes and its care--what do patients expect? AB - A sample of 77% of the non-insulin dependent diabetics aged 30-70 years from two urban practices offering no structured diabetic care were interviewed. The 55 patients (mean age 60 years) were asked about their experiences and expectations of diabetes and the health professionals involved in their care. Twenty-six patients attended the hospital diabetic clinic regularly but 13 patients received no review at all; 46 patients wanted their general practitioner to be involved in future care and only six wanted to continue with hospital review alone. Patients gave hospital doctors and general practitioners similar high ratings for knowledge of diabetes and its management but general practitioners and practice nurses were rated more highly for communication and accessibility. The aspect of care valued most was being given clear information about diabetic management. Twenty two patients thought that diabetes would have a significant impact on their future health and 35 rated regular diabetic review as extremely important in keeping themselves healthy. Most patients felt it likely that they would have a high blood glucose level most of the time and develop diabetic complications. Little difference was found between the views of clinic attenders and non attenders, and there was no evidence that non-attenders had actively rejected review. These non-insulin dependent diabetics considered diabetes to be a serious disorder warranting regular care and expressed confidence in the primary care team's ability to provide such care. PMID- 2556560 TI - Health education using video recordings in a general practice waiting area: an evaluation. AB - With the help of a patient participation group video recordings of health education programmes were shown in the waiting area of a health centre. Patients could choose whether or not to watch the programmes. When asked their views on the project 87% of patients responded positively, and 50% of those who watched could recall specific facts presented. There were no significant differences in age or sex between those who chose and chose not to watch the video recordings. However, unemployed patients and those in social classes 4 and 5 were significantly less likely to watch the programmes than those in social classes 1 3. The use of video recordings in both primary care and hospital waiting areas and the possible role of the health education authority as a central resource is discussed. This system could provide a useful means of extending health education, although further research is needed on the response of different social classes to different forms of health education. PMID- 2556561 TI - Sore throat in family practice: comparison of blood agar throat culture with a rapid enzyme immunoassay test for diagnostic purposes. AB - The Ventrescreen (Ventrex) rapid enzyme immunoassay test for detecting group A streptococcal antigen directly from a throat swab was compared with conventional blood agar throat culture in the diagnosis of beta haemolytic streptococcal infection among 311 patients with a sore throat attending a large suburban Jerusalem primary care clinic. Using the throat culture as the 'gold standard' the Ventrescreen test had a sensitivity of 82%, a specificity of 50%, a positive predictive value of 49%, and a negative predictive value of 82% for beta haemolytic streptococcal infection. These results are not good enough for the test to be considered a reliable substitute for throat culture in such a setting. The negative predictive value, however, supports the use of a negative test result to identify those patients in whom antibiotic therapy could be withheld until the result of their throat culture became available. These conclusions are at variance with recommendations from other studies of similar tests in different population groups, and stress the need for the careful evaluation, especially in primary care clinics, of any such rapid test which claims to be able to replace throat culture in the detection of beta haemolytic streptococcal infection. PMID- 2556562 TI - Needs of elderly people in residential homes: comparison of records held by carers and general practitioners. AB - Fifty patients from one practice who were resident in private nursing homes or residential local authority homes for the elderly (part 3) have been studied. The main carers in the residential homes completed a questionnaire on the care requirements, medical problems and unmet needs of each patient. This information was compared with that available to the general practitioners from the patients' medical notes so that areas of poor communication between the doctor and the main career could be identified. The 11 male patients had a mean age of 82 years and the 39 female patients a mean age of 83 years. Many of the patients had complicated medical problems and were highly dependent on nursing care. Carers were unaware of 34 medical problems among the patients and general practitioners were unaware of care needs in eight patients. Improved communication between general practitioners and the carers in residential homes may benefit patients but proper regard must be given to the privacy and confidentiality of medical information in this setting. PMID- 2556563 TI - History of vocational training for general practice: the 1970s and 1980s. AB - During the 1970s and 1980s, vocational training for general practice developed from experimental beginnings to become established nationwide. Committees were set up nationally and regionally and methods devised for the approval of training practices. Important publications related to the discipline of general practice and to educational methodology appeared. The problems that remain include the absence of a compulsory assessment prior to entering general practice and the low remuneration for course organizers. PMID- 2556564 TI - Clinical psychology: a consultative approach in general practice. AB - General practitioners tend to regard psychologists as an alternative resource for patients. This paper describes an approach to clinical psychology in general practice in which a team of psychologists works with a general practitioner on a consultancy basis. In this way the psychologists and general practitioner work together, with the general practitioner retaining responsibility for the patient. The method pays special attention to the context in which the problems occur, and to the set of relationships in which difficulties are experienced. The relevance of brief focused work is discussed and illustrated with clinical examples. PMID- 2556565 TI - Asian patients' preferences for GPs. PMID- 2556567 TI - Telephone consultations. PMID- 2556566 TI - Dangers of intranasal desmopressin for nocturnal enuresis. PMID- 2556568 TI - GPs' advice to travellers. PMID- 2556569 TI - GPs should not counsel long-term. PMID- 2556570 TI - Importance of legible prescriptions. PMID- 2556571 TI - The community pharmacist. PMID- 2556572 TI - Dispensing by the GP. PMID- 2556573 TI - Failures of screening. PMID- 2556574 TI - Geriatric day hospitals. PMID- 2556575 TI - Sick doctors. PMID- 2556576 TI - Birmingham diabetes survey. PMID- 2556577 TI - Leukotriene receptors. AB - Leukotrienes are metabolites of arachidonic acid catalyzed by a novel lipoxygenase specific for the C-5 position. These fatty acids have many pharmacological properties including smooth muscle contractions and leukocytes activation, and are believed to play a key role in inflammatory and hypersensitivity reactions. Experimental evidence suggests that the action of leukotrienes is mediated by specific receptors. In this paper pharmacological and biochemical data in favor of the existence of leukotriene receptors are presented and hypotheses on the mechanisms of action and transduction are discussed. PMID- 2556578 TI - Heterogeneity between rat and calf peripheral-type benzodiazepine binding sites: differential sensitivity to Triton X-100. AB - The effect of various detergents treatment on the specific binding of [3H]PK 11195 (2nM) to peripheral-type benzodiazepine binding sites (PBS) in calf and rat kidney, adrenal gland, and cerebral cortex membranes was studied. At a concentration of 0.025%, Triton X-100 increased [3H]PK 11195 specific binding to calf kidney, adrenal gland, and cerebral cortex membranes by 20-40%. At the same concentration, Triton X-100 scarcely affected specific binding of [3H]PK 11195 to rat cerebral cortex but decreased binding to rat kidney and adrenal gland membranes by 20-30%. At a concentration of 0.05% of Triton X-100, [3H]PK 11195 specific binding to calf kidney, adrenal gland, and cerebral cortex membranes was increased by 10-20%; whereas [3H]PK 11195 specific binding to rat kidney, adrenal gland, and cerebral cortex membranes was decreased by more than 40%. The increase in [3H]PK 11195 specific binding to calf kidney membranes following Triton X-100 (0.05%) treatment was apparently due to an increase in the binding affinity of PBS, since the density remained unaltered; whereas, the decrease in [3H]PK 11195 specific binding to rat kidney membranes was due to a decrease in both binding affinity and density of PBS. On the other hand, the detergents 3- [(3- cholamidopropyl)- dimethylammonio] - 1 - propane sulfonate (CHAPS), Tween 20, deoxycholic acid, and digitonin have a similar effect on [3H]PK 11195 specific binding to PBS in both calf and rat kidney membranes. PMID- 2556579 TI - Efficacy of different strategies of aftercare for patients who have attempted suicide. PMID- 2556580 TI - Tonsillar metastasis at presentation of small cell carcinoma of the lung. PMID- 2556581 TI - Gating in iodate-modified single cardiac Na+ channels. AB - Elementary Na+ currents were recorded at 19 degrees C during 220-msec lasting step depolarizations in cell-attached and inside-out patches from cultured neonatal rat cardiocytes in order to study the modifying influence of iodate, bromate and glutaraldehyde on single cardiac Na+ channels. Iodate (10 mmol/liter) removed Na+ inactivation and caused repetitive, burst-like channel activity after treating the cytoplasmic channel surface. In contrast to normal Na+ channels under control conditions, iodate-modified Na+ channels attain two conducting states, a short-lasting one with a voltage-independent lifetime close to 1 msec and, likewise tested between -50 and +10 mV, a long-lasting one being apparently exponentially dependent on voltage. Channel modification by bromate (10 mmol/liter) and glutaraldehyde (0.5 mmol/liter) also included the occurrence of two open states. Also, burst duration depended apparently exponentially on voltage and increased when shifting the membrane in the positive direction, but there was no evidence for two bursting states. Chemically modified Na+ channels retain an apparently normal unitary conductance (12.8 +/- 0.5 pS). Of the two substates observed, one of them is remarkable in that it is mostly attained from full-state openings and is very short living in nature; the voltage-independent lifetime was close to 2 msec. Despite removal of inactivation, open probability progressively declined during membrane depolarization. The underlying deactivation process is strongly voltage sensitive but, in contrast to slow Na+ inactivation, responds to a voltage shift in the positive direction with a retardation in kinetics. Chemically modified Na+ channels exhibit a characteristic bursting state much shorter than in DPI-modified Na+ channels, a difference not consistent with the hypothesis of common kinetic properties in noninactivating Na+ channels. PMID- 2556582 TI - Determination of gold-labelled surface receptors on single cells by X-ray microanalysis. AB - A standardless X-ray microanalytical procedure has been developed to determine the number of gold-labelled surface receptors on whole single cells. The effect of the injection of K2PtCl4 into mice on gold-labelled concanavalin A (Con A) receptors on peritoneal macrophages was examined with an energy dispersive X-ray detector in an SEM. The numbers of gold particles seen in electron micrographs and estimated by fluorescence photometric measurements of fluorescein isothiocyanate-labelled Con A receptors were correlated with the X-ray microanalytical results. PMID- 2556583 TI - Ultrathin sectioning for electron microscopy: the distilled water in the knife trough may extract phosphatase reaction products from the sections. AB - During an electron microscopical study of the localization of the nucleoside diphosphatase IDPase in Reissner's membrane of the inner ear, it was discovered that the distilled water in the knife trough produced an annoying artefact. It dissolved all the lead phosphate reaction product from the sections, and thus converted a positive phosphatase reactivity to a false negative one. The water in the knife trough had a pH of approximately 5.4. Calculations showed that this is an expected acidity, if CO2 in the air equilibrates with distilled water, and that there is 200,000 times more acid in the trough than necessary to dissolve all the reaction product from a ribbon of ultrathin sections. Experiments showed that the artefact could be avoided by adjusting the pH to neutrality with dilute ammonia. PMID- 2556584 TI - Clinical correlations of chromosome change in human solid tumors: the tip of the iceberg? PMID- 2556585 TI - Treatment of SCLC: consensus needed. PMID- 2556586 TI - Isochromosome of the short arm of chromosome 12: clinically useful markers for male germ cell tumors. AB - Twenty-nine tumor specimens were obtained from 24 males with germ cell tumors. All primary sites and histologies were represented. An isochromosome of the short arm of chromosome 12 [i (12p)] was found in 20 specimens obtained from 16 patients, a 46,XY normal karyotype was present in seven specimens, and one specimen was a cytogenetic failure. The i(12p) was found in tumors from all primary sites and in all histologies, including a choriocarcinoma. The presence of three or more additional copies of 12p was associated with a statistically significant greater likelihood of treatment failure. With diagnostic and possibly prognostic importance in germ cell tumors in males, the high frequency of i(12p) in our series of studies and in those of others indicates that it probably occurs as a very early defect in the development of these tumors. Further studies of chromosome 12 in males with these tumors are warranted. PMID- 2556587 TI - Phosphatidate phosphohydrolase in purified rat brain myelin. AB - Highly purified myelin from rat brain stem has been shown to contain phosphatidate phosphohydrolase, an enzyme which converts phosphatidate to diacylglycerol. The high levels relative to cytosol and microsomes (17% and 22%, respectively) tended to preclude contamination by these fractions as the source of activity. Additional evidence came from study of repeated purification, mixing experiments, and washing of the myelin with salt and detergent. We conclude that this enzyme, in addition to being widely distributed in other subcellular fractions, is intrinsic to the myelin membrane. Through its activity it generates a key substrate for the cytidine (Kennedy) pathway which was previously shown to occur in this membrane. PMID- 2556588 TI - [A four-year follow-up study of hepatocellular carcinoma with bone metastasis]. AB - A 68-year-old male was admitted to hospital for treatment of a hepatocellular carcinoma with a bone metastasis. By TAE and the administration of PSK, the tumor was necrotized and the AFP level returned to normal range. We presume that this combination therapy of TAE and PSK was able to maintain normal cellular immunity and bring about this good result. PMID- 2556589 TI - [A case of metastatic liver cell carcinoma in the abdominal wall due to direct skin invasion from protruding liver tumor]. AB - A case of a hepatocellular carcinoma with a metastasis to abdominal wall is reported. The patient was a 52-year-old man who, in December, 1982, had been found to have a 16 x 13 cm of tumor in the right lobe of the liver. He received transcatheter arterial embolization (TAE) several times and a hepatic segmentectomy twice. He survived more than six years until his death on January, 17, 1989. A subcutaneous tumor that had appeared in May, 1988 had gradually increased in size and was found to have reached 6 x 5.5 x 2.5 cm on autopsy. The tumor had connected directly with the liver tumor. PMID- 2556590 TI - [Virus-associated hemophagocytic syndrome due to EB virus]. AB - A 30-year-old man was admitted to our hospital because of fever and cervical lymphadenopathy. Hematological examination revealed leukocytosis with atypical lymphocytes in peripheral blood. Mature histiocytes with erythrophagia were detected in the bone marrow. GOT and GPT were elevated. Anti-EB virus antibody titer was high. The titers of VCAIgM and VCAIgG on admission were 1:320 and 1:160, respectively, and those at convalescence stage were 1: less than 10 and 1:640. The diagnosis of virus associated hemophagocytic syndrome (VAHS) due to EB virus was made. Immunosuppressive and cytotoxic therapy are thought to be contraindicated in the treatment of VAHS. In this case only a febrifuge was administrated and the condition of the patient was improved. PMID- 2556591 TI - [CD8+ agranular lymphocyte proliferative disorder with T-cell receptor beta-chain gene rearrangement associated with thymoma and neutropenia]. AB - A 58-year-old male with a 10-years history of thymoma was admitted to our hospital because of the respiratory infection. Hepatosplenomegaly and systemic lymphadenopathy were revealed on physical examination. Chest roentgenogram showed a large anterior mediastinal tumor and a right pleural effusion. Blood examination showed Hb 11.5 g/dl, leucocyte count 1,600/microliters (1% neutrophils, 34% monocytes, 65% lymphocytes) and platelet count 11.2 x 10(4)/microliters. The lymphocytes in the peripheral blood and pleural fluid were mostly small agranular mature lymphocytes and CD2+ 3+ 4- 8+. A monoclonal rearrangement of TCR beta chain gene was found using Southern blot analysis of the lymphocytes in the peripheral blood and pleural fluid. The CFU-GM colony formation by bone marrow cells was normal, and not suppressed by the patient's serum or peripheral blood lymphocytes. Neutrophil-associated IgG was increased with a direct immunofluorescence test. Serum IgG level was slightly decreased. Radiation therapy for thymoma exerted no effect. Treatment with prednisolone 60 mg daily resulted in complete disappearance of the pleural effusion and partial improvement of hepatosplenomegaly, thymoma and neutropenia. Histological examination of the thymoma revealed predominantly spindle cell type. He is still in good condition 21 months after diagnosis. This case seems to represent neoplastic proliferation of mature CD8+ T cells associated with thymoma. PMID- 2556592 TI - [Spontaneous differentiation from myeloperoxidase-negative acute nonlymphocytic leukemia to acute myelomonocytic leukemia]. AB - We reported a 68-year-old woman with acute nonlymphocytic leukemia, in whom the leukemia transformed from poorly differentiated myeloperoxidase (MPO)-negative type into myelomonocytic type during the observation without chemotherapy. Hematological findings on admission revealed a leukocyte count of 3,500/microliters with 48% blasts and a platelet count of 9.2 x 10(4)/microliters. Bone marrow aspiration showed 68.2% infiltration of blasts negative for MPO. Sudan black B and esterase stains. By electron microscopy MPO was detected in the endoplasmic reticulum and nucleoenvelope of the blasts. Large vacuole-like granules were MPO-negative. She was observed without administration of any antileukemic agent or an immunopotentiator. The leukocyte count rose gradually, in association with increases in the relative and absolute counts of mature neutrophils and monocytic cells, and the platelet count. Twenty-six months after the initial diagnosis, a blood examination showed a leukocyte count of 74,300/microliters with 20.5% mature neutrophils and 15.5% monocytic and a platelet count of 31.4 x 10(4)/microliters. Cytological, cytochemical, ultrastructural and immunological studies of the bone marrow cells showed features compatible with acute myelomonocytic leukemia (FAB M4). This case is unusual in respect that poorly differentiated ANLL transformed spontaneously into moderately differentiated ANLL. PMID- 2556593 TI - [Non-pharmacological treatment of hypertension]. PMID- 2556595 TI - [A case of adrenal metastasis of hepatocellular carcinoma effectively treated with local ethanol injection]. AB - We experienced a case of hepatocellular carcinoma with adrenal metastasis that was effectively treated with ultrasonographically guided percutaneous ethanol injection. Effectiveness was proved by various imaging procedures and decreased serum alpha-fetoprotein. No complications during and after the treatment were deserved. PMID- 2556596 TI - [Adrenal arterial embolization and radiotherapy of a case with metastatic adrenal tumor and its tumor thrombus in inferior vena cava from hepatocellular carcinoma]. AB - Right adrenal arterial embolization and radiotherapy were performed in a patient to treat the metastatic tumor and its tumor thrombus in inferior vena cava from hepatocellular carcinoma. This case developed no complication following the embolization and radiotherapy. This therapy necrosed the metastatic tumor and made inferior vena cava recanalization. Adrenal arterial embolization and radiotherapy were effective treatment for the metastatic adrenal tumor and its tumor thrombus. PMID- 2556594 TI - [Endothelin: a novel endothelium-derived peptide]. PMID- 2556597 TI - [Imaging of adenomatous hyperplastic lesions containing and not containing hepatocellular carcinoma in the liver]. AB - In 41 patients with 54 lesions which were resected ans studied histopathologically, there were 14 lesions of adenomatous hyperplasias (AH) in 9 patients, 28 AHs containing hepatocellular carcinoma foci (early HCC, e-HCC) in 22 and 12 borderline lesions which fell between these two lesions in 10. The detectability of these lesions on imagings was evaluated. Detection rates for all lesions and e-HCCs were as follows; intraoperative sonography, 70.0%, 87.5%; Portal-CT, 71.4%; sonography, 44.4%. 64.3%; Arterial-CT, 37.5%, 50.0%; CT, 32.7%, 57.7%; angiography, 17.0%, 30.8%; Lipiodol-CT, 9.1%. 25.0%. On angiography, tumor stain was recognized in only 8 patients with e-HCC. Arterial-CT showed a relatively low density mass compared to non-tumorous area in 2 patients with e HCC and one with borderline lesion. The median size of 54 lesions was 1.2 +/- 0.4 cm in diameter and that of AHs was 0.8 +/- 0.3 cm, the latter being significantly smaller than the other two lesions (p less than 0.01). Liver cirrhosis coexisted in 35 of 41 patients (85.4%). No complete necrosis occurred in 13 e-HCC lesions following therapeutic embolization or infusion chemotherapy in the hepatic artery. PMID- 2556598 TI - [Localization of pancreatic insulinomas by combined radiographic methods]. AB - Seventeen cases with a total of eighteen insulinomas were examined by ultrasonography (US), computed tomography (CT), angiography, percutaneous transhepatic portal and pancreatic venous sampling (PTPS) and intraoperative ultrasonography (IOUS). We discussed about diagnostic accuracy of insulinomas on these diagnostic modalities. Tumor localization was achieved by US in 39%, CT in 72% and angiography in 81%. Although total 15 tumors (83%) can be correctly detected, multiple lesions were not completely ruled out. We recommended the criteria in PTPS as follows: Insulin concentration level above 200 mu U/ml. Or, both of insulin concentration ratio (step-up site/control) above three and elevation of the CPR concentration level on the PTPS. According to these criteria, the accuracy rate was 91%. Overall preoperative diagnostic accuracy was 100%. IOUS allowed us to detect the localization of the tumors completely. PMID- 2556599 TI - [Carcinoma of the papilla of Vater associated with pancreatolithiasis--a case report]. PMID- 2556600 TI - [An autopsy case of hepatocellular carcinoma with tumor thrombus in the right atrium]. PMID- 2556601 TI - [An autopsy case of hepatocellular carcinoma invaded into ligamentum teres hepatis]. PMID- 2556602 TI - [Imaging diagnosis of ruptured site in hepatocellular carcinoma]. PMID- 2556603 TI - "Radical" abdominal cancer surgery: current state and future course. AB - Biologically driven cancer surgery is essential for the best care of the cancer patient. Modern technology and support have improved the safety and operative results of radical surgery. Better understanding of tumor biology with technical advances permit rational, en bloc resection of the primary cancer and regional lymphatic drainage with better cure rates. The author has operated on 1103 patients with liver tumors since 1970. The liver tumors were resected in 415 patients, with major hepatic resections being done in 359. Since 1979, the author carried out 62 regional pancreatectomies. Thirty-three were for stage 1 or 2 adenocarcinomas of the head of the pancreas. The 5-year actual survival was 31 per cent. There has been similar improvement in treating ampullary and periampullary cancer, gallbladder cancer, or extrahepatic bile duct cancer. These data show significant improvement in the results of treating cancer by radical or biologically driven surgery. Abdominal cancer is not a generalized disease until late in its course. PMID- 2556604 TI - Multicystic kidney in association with hepatoblastoma--a case report. AB - In the absence of functional renal tissue on the right side, differentiating between a liver tumor and a renal tumor, both clinically and on routine imaging, is difficult. This clinical dilemma is highlighted here in a case report of a 2 year old girl with hepatoblastoma occurring in association with multicystic kidney. To our knowledge no other such case has been reported so far. PMID- 2556605 TI - [Induction of differentiation in testicular choriocarcinoma cells and its role in cancer therapy]. AB - As an approach to develop a new therapeutic modality for testicular germ cell tumor, in vitro studies were performed to investigate the capacity of testicular choriocarcinoma cell line JHTK-1 to differentiate and to screen its differentiation inducing substances. We have found that dibutyryl cyclic adenosine monophosphate (DBcAMP) induces multiple phenotypic changes of JHTK-1 cells within 48hr, i.e., morphological alteration, enhancement of human chorionic gonadotropin (HCG) production, and inhibition of the cellular proliferation. Upon addition of 1 mM DBcAMP to the culture mediums, broad and elongated cells with cytoplasmic projections, increased secretion of HCG to the culture mediums more than 15-fold and increased number of HCG positive cells more than 10-fold in immuno-histochemical study, and inhibition of cell proliferation more than 50 percent were seen. The changes were dose dependent as to DBcAMP. These results indicate that DBcAMP induces the differentiation of testicular choriocarcinoma cells from dividing cytotrophoblastic cells to non-dividing end cells of syncytio trophoblastic cells. Screening of differentiation inducing substances were carried out using an increase in HCG secretion/cell/48 hr as an indicator of JHTK 1 cell differentiation. The inducers were theophylline, sodium butyrate, N butyrate, vitamin C, vinblastine, and actinomycin D. In conclusion external stimuli such as chemical substances can induce the differentiation of testicular choriocarcinoma cells, and inhibit their cellular proliferation as well. This finding will lead to a concept of "differentiation induction therapy" for testicular choriocarcinoma. PMID- 2556606 TI - [Functional roles of cyclic AMP and cyclic GMP on rabbit urinary bladder and urethral smooth muscle contraction]. AB - It is suggested that cyclic AMP and cyclic GMP play regulatory roles in smooth muscle function. In order to clarify the role of cyclic AMP and cyclic GMP in the function of the lower urinary tract, I examined the effects of isoproterenol and sodium nitroprusside on contractile force and tissue content of cyclic AMP and cyclic GMP. Isoproterenol (10(-8)-10(-4) M) caused a relaxing effect in the rabbit urinary bladder dome, the bladder base and the urethra. The relaxation response by isoproterenol was mediated by cyclic AMP. The relaxation response and the increase in the cyclic AMP level were marked in the bladder dome. Sodium nitroprusside (10(-8)-10(-4) M) also caused a relaxing effect in the rabbit urinary bladder dome, the bladder base and the urethra. The relaxation response to sodium nitroprusside was mediated by cyclic GMP. The relaxation response and the increase in the cyclic GMP level were marked in the urethra. The external addition of derivatives of cyclic AMP and cyclic GMP (dibutyryl cyclic AMP and 8 bromo cyclic GMP, 10(-8)-10(-3) M) also caused a relaxing effect in the bladder dome, the bladder base and the urethra. These results demonstrated that both cyclic AMP and cyclic GMP were related to relaxation of the rabbit lower urinary tract smooth muscle. It seems that cyclic AMP may be mainly related to relaxation of the bladder dome and cyclic GMP may be mainly related to relaxation of the urethra. PMID- 2556608 TI - [2 rare tumors of the esophagus]. PMID- 2556607 TI - Abnormal norepinephrine uptake and release in brain synaptosomes in chronic renal failure. AB - Abnormalities in the function of the central nervous system exist in chronic renal failure (CRF) and some of these derangements may be related to excess parathyroid hormone (PTH) which causes a rise in brain calcium. The latter may affect metabolism of neurotransmitters such as norepinephrine (NE) in brain synaptosomes. We measured NE content, uptake and release in brain synaptosomes of CRF rats and studied whether excess PTH affects these parameters. Synaptosomes from rats with 21 days of CRF compared to those from normal animals have higher calcium content (11.4 +/- 0.92 vs. 7.1 +/- 0.50 nmol/mg protein, P less than 0.01) and lower Na-K ATPase activity (6.5 +/- 0.81 vs. 11.4 +/- 0.76 mumol Pi/mg protein/hr, P less than 0.01). NE content (11.0 +/- 0.60 vs. 13.6 +/- 0.55 pmol/mg protein/hr, P less than 0.01), uptake (46 +/- 4.5 vs. 110 +/- 5.9 pmol/mg protein times 50 min, P less than 0.01) and release (2.0 +/- 0.2 vs. 5.1 +/- 0.47 pmol/mg protein times 10 min, P less than 0.01). Parathyroidectomy (PTX) in CRF rats kept normocalcemic reversed these abnormalities in brain synaptosomes; indeed calcium content, Na-K ATPase activity and NE content, uptake and release in synaptosomes from PTX-CRF rats were not different from those seen in normal rats.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2556609 TI - Thymic necrosis following oral inoculation of mouse thymic virus. AB - Mouse thymic virus (MTLV;ICTV designation murid herpesvirus 3) infects developing T lymphocytes of neonatal mice, causing thymic necrosis and acute immunosuppression. Infected animals shed virus indefinitely. However, although transmission in nature is presumably by contact and is likely to involve the oral nasal route, virtually all experimental studies with MTLV have used systemic (intraperitoneal) inoculation. In order to determine whether systemic inoculation causes artifacts in pathogenesis of the infection, effects of intraperitoneal and oral-nasal inoculation were compared in newborn mice. Thymic necrosis occurred with either route of inoculation, although rate of infection was lower with oral inoculation, varying from about 20% to 67%. There were no gross differences in pathogenesis. Orally infected animals seroconverted and shed virus. These data indicate that the apparent lymphotropism of thymic virus, and induction of thymic necrosis, are not dependent on route of inoculation. PMID- 2556610 TI - Perinatal diagnosis of type 1c glycogen storage disease. PMID- 2556612 TI - Detection of Fabry's disease carriers by enzyme assay of hair roots. PMID- 2556611 TI - Tyrosinaemia type I: orthotopic liver transplantation as the only definitive answer to a metabolic as well as an oncological problem. PMID- 2556613 TI - Isolated breast metastases from gastrointestinal carcinomas: report of two cases. AB - Two patients with metastatic gastrointestinal adenocarcinoma in the breast are reported. Metastases in the breast are usually painless upper outer quadrant masses. On mammography they are typically well-circumscribed lesions without microcalcifications. A breast mass in a patient with a history of cancer, even if clinically or mammographically benign, should raise suspicion of a metastasis. Pathological features include a histologic pattern similar to the primary neoplasm and an absence of in situ carcinoma which characterizes primary breast cancer. Surgical excision for local control and systematic therapy is the most appropriate treatment. PMID- 2556615 TI - Gestational alterations in phospholipase C coupled muscarinic response. AB - In the pregnant rat, carbachol-induced phosphoinositol hydrolysis by myometrium at the placental attachment region progressively decreased toward term, whereas hydrolysis was relatively stable in the myometrium of the non-attachment region. Tritium-quinuclidinyl benzilate binding increased in the myometrium of non attachment regions as pregnancy progressed. At placental attachment sites binding remained relatively stable until parturition when it increased. Apparently the myometrium associated with the placental attachment site is less sensitive to cholinergic influence during pregnancy compared with the non-attachment site when evaluated by muscarinic activation of phospholipase C or ligand binding. PMID- 2556614 TI - Dextromethorphan and sigma ligands: common sites but diverse effects. AB - There is increasing evidence that sigma ligands and dextromethorphan (DM) bind to at least one common high-affinity site. DM and other antitussives do not produce psychotomimetic effects. This suggested that sigma ligands may produce their characteristic effects through another site, and prompted us to review critically the literature on the side effects of sigma opiates. Contrary to what is generally accepted, the dysphoric and psychotomimetic side effects of sigma opiates are mediated by the levo-and not by the dextrorotatory isomers. Moreover, these effects are unequivocally naloxone-reversible. Therefore, the current version of the "sigma receptor", with high affinity for the dextrorotatory sigma opiates, cannot explain the psychotomimetic effects of the levorotatory enantiomers. Thus, neither the "sigma ligands" nor its newly defined "receptor" are involved in the psychotomimetic effects of sigma opiates. Further experimentation with more selective drugs and with a combination of different methods will be necessary to identify the different binding sites, and to establish their physiological role and therapeutic potential. PMID- 2556616 TI - A new alcohol antagonist: phaclofen. AB - The ability of the GABA(B) receptor antagonist, phaclofen to alter behavioral effects of ethanol was evaluated by loss of righting reflex (sleep time), motor incoordination (bar holding), spontaneous locomotion (open field activity) and hypothermia. Pretreatment with phaclofen significantly decreased the effects of ethanol on motor incoordination, locomotor activity and hypothermia. However, phaclofen had no effect on either pentobarbitalor diazepam-induced motor incoordination. Phaclofen slightly increased the ED50 for loss of the righting reflex but did not alter either the duration of reflex loss produced by ethanol or blood ethanol levels at awakening. Our results suggest phaclofen is rapidly inactivated resulting in difficulty in observing antagonism of long duration ethanol effects. These findings suggest that the GABA(B) system may play a role in mediating several important actions of ethanol. PMID- 2556617 TI - Effects of the selective kappa opioid antagonist, nor-binaltorphimine, on electrically-elicited feeding in the rat. AB - Lateral ventricular injections of the 'nonspecific' opioid antagonist naloxone (100 micrograms) and the kappa-selective opioid antagonist nor-binaltorphimine (50 micrograms) elevated the electrical brain stimulation frequency threshold for eliciting feeding behavior. Mesopontine aqueductal injections of nor binaltorphimine, on the other hand, lowered the feeding threshold while naloxone still elevated threshold. These findings suggest the existence of forebrain kappa receptors at which endogenous opioid activity results in a facilitation of feeding while kappa receptors in the brainstem seem to mediate an inhibitory effect. PMID- 2556618 TI - Expression of NGF receptor in the rat forebrain detected with in situ hybridization and immunohistochemistry. AB - The expression of nerve growth factor (NGF) receptor mRNA and NGF receptor protein was examined in the adult rat basal forebrain using in situ hybridization and immunohistochemical techniques. NGF receptor mRNA and protein were detected within cells in the medial septum, diagonal band of Broca, and nucleus basalis of Meynert. Controls showed that the hybridization signal was not due to nonspecific binding of the probe to heterologous RNAs or other molecules. As expected, the distribution of NGF receptor mRNA-containing cells correlated nicely with the distribution of NGF receptor immunoreactive cells in each of these areas. These data extend previous work which suggests that neurons in these areas express the NGF receptor mRNA and manufacture functional NGF receptors. NGF receptor immunoreactivity was also detected in the arcuate nucleus of the hypothalamus, in the leptomeninges at the base of the brain and overlying the tectum, and within ependymal regions along the lateral walls of the cerebral ventricles. A few weakly stained neurons in the lateral hypothalamus and ventrolateral striatum were also consistently observed. In contrast, NGF receptor mRNA was not detected within any meningial, ependymal, or hypothalamic tissues using in situ hybridization. A cross-linking/immunoprecipitation assay demonstrated normal, membrane-bound NGF receptors within extracts of dorsal superior colliculus, ventromedial hypothalamic, and overlying meningial tissues, proving that the staining observed in these areas was not a non-specific artifact associated with the immunohistochemistry. The lack of hybridization in these areas may reflect levels of NGF receptor mRNA which are too low to be detected by the in situ hybridization methods being used. Alternatively, the staining may represent innervation of these areas by afferents whose cell bodies are located elsewhere, and whose terminals contain the NGF receptor protein. PMID- 2556619 TI - Molecular and biological characterization of a type 1 herpes simplex virus (HSV 1) specifically deleted for expression of the latency-associated transcript (LAT). AB - We have previously reported that in sensory neurons of mice harboring latent herpes simplex virus (HSV), a novel virus-encoded RNA species termed the latency associated transcript (LAT) is abundant. To investigate a potential function for LAT in the latent state, we have engineered a deletion in HSV-1 strain KOS(M) which abolishes LAT expression. The deletion virus (KOS 8117) is not altered for virulence or the ability to replicate in mouse cells in vivo or in vitro. Although no transcripts were detected in lumbosacral ganglia of mice after virus inoculation on scarified rear footpads, the deletion virus established a latent infection since infectious virus could be recovered by explanting and cultivating ganglia in vitro with permissive cells. These results indicate that expression of LAT is not an absolute requirement for the establishment or maintenance of the latent state or for recovery of infectious virus from the latent state by in vitro co-cultivation. Since no viral-encoded transcripts were detected in cells latently infected with KOS 8117, maintenance of the latent infection also appears to be exclusively a neuronal function. PMID- 2556621 TI - Molecular cloning and analysis of the incompatibility and partition functions of the virulence plasmid of Salmonella typhimurium. AB - The incompatibility functions (inc) of the virulence plasmid of Salmonella typhimurium LT2 were initially located in a 4.3 kb region near the repA locus of the plasmid. Expression of inc required the presence, in cis or in trans, of two distinct DNA regions of the fragment. These regions, maximally 0.3 kb and 0.6 kb in size, were separated in the fragment by c. 3.0 kb. This intervening DNA encoded two proteins, of Mr values 37 kDa and 40 kDa. The promoter for the 37 kDa protein lay in or near one of the inc regions (incL). No function was assigned to this protein, however, it may be the product of a rep gene. The 40 kDa protein may have a partition (par) function, and may bind to a centromeric site in or near the other inc region (incR). An inc+ par- derivative of the original plasmid clone was used in a simple method, not involving the use of curing agents/mutagens, to eliminate virulence plasmid DNA from Salmonella typhimurium, Salmonella dublin, Salmonella enteritidis, and Salmonella cholerae-suis. The par function served to stabilise pJRD158B-based plasmid greater than 10(6)-fold in Escherichia coli and the virulence plasmid-cured Salmonella strains. PMID- 2556620 TI - Increased monokines in cytomegalovirus infected myelomonocytic cell cultures. AB - In an investigation of the role of monokines in CMV associated immunosuppression we document modulation of both TNF and IL-1 beta in CMV infected THP-1 cells. CMV infected cultures released almost two-fold more IL-1 beta protein and contained significantly higher IL-1 beta mRNA levels than uninfected cultures for 72-96 h after induction. In both CMV infected and uninfected cultures, significant amounts of IL-1 beta protein were not detected until 24 h post induction, while maximum amounts of TNF were detected in culture supernatants by 3 h post induction, suggesting that TNF may play a role in IL-1 beta induction. TNF levels subsequently declined but in infected cultures remained over 2.5-fold higher than controls through 96 h. The CMV alteration in the kinetics and extent of IL-1 beta release must be indirectly mediated by CMV since only 1% of THP-1 cells were infected. Most infected cells expressed CMV immediate early proteins but did not overexpress IL-1. We speculate that CMV infected cells release excess TNF or other stimulatory factors which increase IL-1 beta synthesis. Since IL-1 beta is increased, the decreased IL-1 'activity' described by others as an explanation in part for the immunosuppressive effects of infection may actually reflect alterations of IL-1 inhibitor levels during CMV infection. PMID- 2556622 TI - Effects of sodium bicarbonate on nitrogen metabolism and ketone bodies during very low energy protein diets in obese subjects. AB - This study evaluated the effects of oral sodium bicarbonate (NaHCO3) supplementation on ammonium (NH4+) nitrogen (N) and urea N excretion and on ketone bodies during the metabolic acidosis of a very low energy protein diet. Ten healthy obese female subjects (BMI, 38.4 +/- 1.5 kg/m2;weight, 100 +/- 4 kg) were given a 1.72 MJ (412 kcal) all protein (16.8 g N) liquid formula, 16 mmol KCl and a multivitamin-mineral supplement daily for 4 weeks. In addition, the five subjects in group 1 received 60 mmol Na+ daily as sodium chloride (NaCl) for 3 weeks and as NaHCO3 during week 4. The subjects in group 2 were given 40 mmol/d NaHCO3 during the first week, 60 mmol/d during weeks 2 and 3, and 60 mmol/d NaCl during week 4. Nitrogen balance was achieved in both groups by the end of week 3. The subjects in group 1 at week 2 showed an increase in blood [H+] of 0.41 +/- 0.06 x 10(-8) mol/L and a decrease in blood bicarbonate from 26.0 +/- 0.8 to 23.8 +/- 1.2 mmol/L. The subsequent NaHCO3 curtailed NH4+ N excretion by one half, without significant change in ketone body levels or excretion. Administration of NaHCO3 from the start of the diet to the subjects in group 2 prevented both the metabolic acidosis and the increase in NH4+ N excretion and attenuated the increase in blood and urine 3-hydroxybutyrate. When NaCl replaced NaHCO3 during week 4, ammonium N excretion doubled. Urea N excretion was comparable in both groups and was unaffected by bicarbonate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556623 TI - Determination of surface potential of biological membranes. PMID- 2556625 TI - Cell separation using velocity sedimentation at unit gravity and buoyant density centrifugation. PMID- 2556624 TI - Cell isolation techniques: use of enzymes and chelators. PMID- 2556626 TI - Use of ionophores in epithelia: characterizing membrane properties. AB - In this chapter we have described the use of antibiotics for studying epithelial membrane resistances and permeabilities, as well as active and passive transport processes. We placed particular emphasis on sources of artifacts that could result in erroneous determination of any of the above parameters, as well as methods for assessing such artifacts (where possible). Although the number of possible artifacts seems large, it is not our intention to dissuade researchers from using this probe of epithelial function, but rather to point out how data might be incorrectly interpreted. PMID- 2556627 TI - Implantation of isolated carriers and receptors into living cells by Sendai virus envelope-mediated fusion. PMID- 2556628 TI - Catecholamine-mediated inactivation of poliovirus RNA. AB - Catecholamine (DOPA, dopamine, epinephrine, or norepinephrine) and Cu2+ produced a potent inactivator(s) of transfectivity of poliovirus RNA. Al3+, Co2+, Fe3+, Mn2+, Ni2+, Sn4+ and Zn2+ did not substitute for Cu2+. Potency of inactivator(s) was eliminated by chelator (EDTA, histamine, or L-histidine). Kinetics of inactivation was single-hit, and inactivation was first order with respect to inactivator(s). A method for estimating relative concentration of inactivator(s) showed its course of production at 23 degrees C from norepinephrine at 10 microM and Cu2+ at 1.0 microM: under air, inactivator(s) peaked after 95 min, then decayed; under oxygen, peak was after 40 min; and under nitrogen, no production was detected. After prolonged exposure, most of the tritium from tritiated viral RNA appeared in small fragments. As tested with Cu2+ and norepinephrine, Al3+ and Fe3+ interfered strongly with production of inactivator(s) but not with preformed inactivator(s). An inactivator role for copper complexes of the semiquinone derivatives is hypothesized. PMID- 2556629 TI - Prevalence of cytomegalovirus antibodies in Brazilian and Japanese populations in the north-east of Brazil. AB - The serological status to cytomegalovirus (CMV) was examined for 616 Brazilians and 399 Japanese immigrants living in the North-East of Brazil. The sera were screened for IgG antibodies to CMV by enzyme-linked immunosorbent assay (ELISA). The overall prevalence of CMV antibodies was higher in the Japanese population (83.7%) than in the Brazilian population (69.8%). The seropositivity was analyzed by factors of age, sex, ethnic background, and socioeconomic status. Mother-baby contact seems to be a factor of significance in the seroepidemiology of CMV infection. PMID- 2556630 TI - The prevention of colorectal cancer in Australia and the advent of registers for familial adenomatous polyposis. PMID- 2556631 TI - Impact of a hospital-based register on the management of familial adenomatous polyposis. AB - Colorectal cancer in familial adenomatous polyposis is a preventable disease in at-risk relatives of patients with primary cases. Until the recent establishment of a register in Western Australia, there has been no registration of pedigrees or central organization of surveillance in Australia. In the present study, the experience of 20 such families who were associated with The Royal Melbourne Hospital was documented, with an analysis of the reasons for any failure of management. The impact of a hospital-based register on the management of the disease was studied. In each family, results were categorized according to whether "at-risk" relatives had been diagnosed at surveillance examinations, and whether the Hospital register were involved. Before involvement with the register, 24 family members presented with symptomatic polyposis after the first affected case had been diagnosed. Eighteen of these had colorectal cancer at diagnosis, and 16 subjects now are dead. Identifiable reasons for the failure of surveillance were family communication failure (two cases), family denial (two cases), failure of the hospital clinic (two cases) and a failure to cover extended branches of families who were living locally (nine cases), interstate (four cases) or overseas (three cases). In contrast, only two (6%) of 33 affected cases that were identified at a planned surveillance endoscopy had colorectal cancer at diagnosis. Without the active surveillance of at-risk family members, lethal delays in diagnosis are likely to occur. Most reasons for failure potentially are correctable by a dedicated registry that is responsible for notifying clinicians and patients about the timing of surveillance procedures. PMID- 2556632 TI - The Western Australian Familial Polyposis Registry. AB - The Western Australian Familial Polyposis Registry was established in 1985 by the Cancer Foundation of Western Australia in conjunction with the Health Department of Western Australia. The primary aim of the Registry is the prevention of colorectal cancer in a group of persons at high risk of the disease. Families are notified to the Registry by medical practitioners throughout the State and, at the request of family members and medical practitioners, the project officer for the Registry will arrange for the collection of data, counselling and reminders for follow-up examinations for both cases and family members who are at risk of developing the disease. A total of 44 families with familial adenomatous polyposis is known to the Registry. The crude incidence of polyposis in Western Australia was 0.24 cases per 100,000 persons over the three-year period 1986 1988. The prevalence of the condition in 1988 was 2.59 cases per 100,000 persons. There are 188 family members in Western Australia who are at a 50% risk of developing familial adenomatous polyposis. Registry data indicate that regular follow-up and early treatment reduce the risk of the development of bowel cancer. PMID- 2556633 TI - Phenytoin toxicity induced by fluconazole. PMID- 2556634 TI - Recovery of viruses and bacteria in waters off Bondi beach: a pilot study. AB - A pilot study was conducted between February and April, 1989, on the occurrence of sewage-derived viruses and bacteria in the beach and nearshore waters off Bondi, Sydney. Enteroviruses were isolated from 41% of a total of 66 sewage, sea water, grease and sediment samples. Poliovirus vaccine strains accounted for 78% of the isolates. Adenoviruses were isolated four times and coxsackievirus B was isolated twice in samples that were collected away from the bathing area. Rotavirus and hepatitis A virus were not detected. The number of faecal coliforms, faecal streptococci, Aeromonas hydrophila and Clostridium perfringens in the sewage ranged from 10(4) to 10(7) cfu/100 mL. An initial reduction of 10(2)-10(3) in bacterial counts was observed in the plume (the effluent's initial dilution zone) and a further reduction of 10(2)-10(4) in counts in samples that were collected away from the plume. Salmonella spp. were detected in one-third of the samples and low numbers of Campylobacter jejuni were found in the sewage and plume. Salmonella serotypes that were associated with the sewage and plume were distinct from those in the stormwater and beachwater, indicating that stormwater may be an important source of pollution on Bondi beach. PMID- 2556637 TI - [Experimental study of injury on the small intestine in acute portal vein occlusion and the following restoration of portal vein flow in rats--free radicals in the small intestine and lipid peroxidation]. AB - Free radicals in the small intestine were quantified by using an electron spin resonance spectrometer, and the amounts of TBA (thiobarbituric acid) reactants in arterial plasma, portal venous plasma and intestinal tissue were determined at the several stages. The effects of allopurinol, alpha-tocopherol, the simultaneous occlusion of superior mesenteric artery or the porto-jugular venous bypass, with the temporary occlusion of the portal vein, were also investigated. 1) Free radical concentration (mostly, semiquinones of CoQ and/or flavin in mitochondria) decreased with portal vein occlusion but showed a temporary increase at 10 sec after reperfusion. Allopurinol suppressed such temporary increase. 2) TBA reactants increased with the temporary occlusion of the portal vein. TBA reactants decreased during the portal vein occlusion with alpha tocopherol and during reperfusion with allopurinol. Lipid peroxidation in the small intestine was also diminished by using the methods of simultaneous occlusion of the superior mesenteric artery or the porto-jugular venous bypass. In conclusion, there may be three sources for the generation of superoxide: the xanthine oxidase system, semiquinone radicals and paramagnetic metal irons. They may induce lipid a peroxidation, which accelerates the injury on the small intestine, in acute portal vein occlusion and the following restoration of portal vein flow in rats. PMID- 2556638 TI - Interleukin-2-induced decrease in the buoyant density of cytotoxic cells and its relationship with proliferation. AB - Interleukin 2 (IL-2) is known to induce an augmentation of natural killer activity. In the present study we have used discontinuous Percoll density gradients to investigate the changes in the buoyant density of killer effector cells generated in response to IL-2. In all systems examined (mouse and rat spleen cells and human peripheral blood mononuclear cells), the cytolytic effectors generated in response to IL-2 have markedly lower buoyant densities compared to fresh natural killer cells. Our results also suggest that if a single layer of 44.2% (v/v) Percoll is used, almost all IL-2-induced cytolytic activity can be enriched in the cells which float on this layer whereas the heavier cells from the pellet are devoid of cytotoxic activity. The contribution of proliferative activity to (a) the generation of cytotoxicity and (b) the decrease in the buoyant density of the IL-2-induced killer cells was also studied in the mouse system. Natural killer levels in mouse spleen cells treated with mitomycin C could be significantly augmented by IL-2. Moreover, the effector cells generated in control as well as mitomycin-C-treated spleen cells in response to IL-2 had the same low buoyant densities. These results indicate that proliferation is not a prerequisite for the activation of killer activity and a reduction in buoyant density in response to IL-2. PMID- 2556639 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 51-1989. A 72-year-old woman with bronchospasm and a pleural effusion after treatment of breast carcinoma. PMID- 2556640 TI - Sequence and expression of a frog brain complementary DNA encoding a kainate binding protein. AB - Excitatory amino acids (EAAs) are important neurotransmitters in the vertebrate central nervous system. Electrophysiological and ligand-binding studies indicate that at least three different receptor subtypes for EAAs exist--N-methyl-D aspartate, kainate and quisqualate receptor subtypes--on the basis of the preferred agonist of the receptors. We recently purified a kainate-binding protein (KBP) from frog (Rana pipiens berlandieri) brain by domoic acid (a high affinity kainate analogue) affinity chromatography, and showed that the kainate binding activity was associated with a protein of relative molecular mass 48,000 (Mr 48 K). The pharmacological properties and the anatomical distribution of KBP were consistent with those of a kainate receptor-ionophore complex. We have now isolated a complementary DNA encoding KBP of Mr 48 K. The deduced amino-acid sequence of the KBP has similar hydrophobic profiles to those found in other ligand-gated ion channel subunits, and shows some amino-acid sequence similarities to the corresponding regions of brain nicotinic acetylcholine receptor subunits. Localization of the KBP messenger RNAs by in situ hybridization histochemistry is compatible with the results of immunohistochemistry and receptor autoradiography studies. COS-7 cells transfected with the cDNA encoding the KBP show high-affinity kainate-binding activity with pharmacological properties similar to those of the biochemically purified KBP. These results provide the first molecular characterization of an EAA-binding site and raise the possibility that the KBP cDNA encodes a ligand binding subunit of a kainate receptor-ionophore complex. PMID- 2556641 TI - The colony stimulating factor-1 receptor associates with and activates phosphatidylinositol-3 kinase. AB - Colony stimulating factor-1 (CSF-1) is a lineage-specific growth factor required for proliferation and survival of mononuclear phagocytes and their precursors. The CSF-1 receptor belongs to a family of ligand-activated protein-tyrosine kinases. Activation of the platelet-derived growth factor receptor, but not the CSF-1 receptor, leads to an increase in phospholipase C activity and a subsequent elevation in intracellular calcium. Recent studies have shown that a novel phosphoinositol (PtdIns) kinase, termed PtdIns-3 kinase, is stimulated by the platelet-derived growth factor receptor and certain oncogenes in the protein tyrosine kinase family. PtdIns-3 kinase phosphorylates the D-3 hydroxyl position of the inositol ring of PtdIns, and its products do not participate in the generation of the second messenger inositol 1,4,5-trisphosphate (Ins(1,4,5)P3). Here we report that addition of CSF-1 is followed by activation of PtdIns-3 kinase in a macrophage cell line (P388 D1), which contains CSF-1 receptors, and in BALB/c fibroblasts made to express the human CSF-1 receptor. Furthermore, we show that activation of the CSF-1 receptor results in the accumulation in intact cells of polyphosphoinositides phosphorylated at the D-3 position of the inositol ring. Thus activation of the CSF-1 receptor stimulates PtdIns-3 kinase activity, indicating a novel pathway for CSF-1 receptor-mediated signal transduction. PMID- 2556643 TI - Sequence-specific RNA binding by the HIV-1 Rev protein. AB - The human immunodeficiency virus type 1 (HIV-1) Rev protein acts post transcriptionally to increase the amounts of the viral gag-pol and env messenger RNAs in the cytoplasm of infected cells. The mechanism of Rev action is uncertain. Possibilities include an accelerating effect on the rate of export of its mRNA targets from the nucleus and/or modulation of the splicing of pre-mRNAs. Both the gag-pol and env mRNAs contain a sequence that is required for responsiveness to Rev--the Rev responsive element, RRE. Here we show that Rev is a sequence-specific binding protein, whose binding site is the RRE. This information should help to clarify the mechanism by which Rev acts. PMID- 2556642 TI - Differential expression of genes encoding alpha, beta and gamma retinoic acid receptors and CRABP in the developing limbs of the mouse. AB - Retinoic acid has profound effects on vertebrate limb morphogenesis (refs 1-6, reviewed in refs 7-9), including in the mouse, where it can act as a teratogen generating phocomelia and bone defects. A retinoic acid gradient, possibly amplified by a graded distribution of cellular retinoic acid-binding protein (CRABP), could provide positional information across the antero-posterior axis of the chick limb bud. The discovery of nuclear retinoic acid receptors (RARs) acting as retinoic acid-inducible enhancer factors provided a basis for understanding how retinoic acid signals could be transduced at the level of gene expression. We have now used in situ hybridization to study the distribution of messenger RNA transcripts of the three murine receptors (mRARs) and CRABP during mouse limb development. Both mRAR alpha and mRAR gamma transcripts, but not those for mRAR beta, are present and uniformly distributed in the limb bud at day 10 post-coitum, whereas CRABP transcripts have a graded proximo-distal distribution, indicating that differential expression of CRABP, but not of mRAR alpha or mRAR gamma, could participate in the establishment of the morphogenetic field. At later stages, mRAR gamma transcripts become specific to the cartilage cell lineage and to the differentiating skin and mRAR beta transcripts are mostly restricted to the interdigital mesenchyme. CRABP transcripts, however, are excluded from regions expressing mRAR gamma and mRAR beta. These results indicate that all three RARs and CRABP have specific functions during morphogenesis and differentiation of the mouse limb. PMID- 2556644 TI - A protein binding to the J kappa recombination sequence of immunoglobulin genes contains a sequence related to the integrase motif. AB - Site-specific recombination requires conserved DNA sequences specific to each system, and system-specific proteins that recognize specific DNA sequences. The site-specific recombinases seem to fall into at least two families, based on their protein structure and chemistry of strand breakage. One of these is the resolvase-invertase family, members of which seem to form a serine-phosphate linkage with DNA. Members of the other family, called the integrase family, contain a conserved tyrosine residue that forms a covalent linkage with the 3' phosphate of DNA at the site of recombination. Structural comparison of integrases shows that these proteins share a highly conserved 40-residue motif. V (D)-J recombination of the immunoglobulin gene requires conserved recombination signal sequences (RS) of a heptamer CACTGTG and a T-rich nonamer GGTTTTTGT, which are separated by a spacer sequence of either 12 or 23 bases We have recently purified, almost to homogeneity, a protein that specifically binds to the immunoglobulin J kappa RS containing the 23-base-pair spacer sequence. By synthesizing probes on the basis of partial amino-acid sequences of the purified protein, we have now isolated and characterized the complementary DNA of this protein. The amino-acid sequence deduced from the cDNA sequence reveals that the J kappa RS-binding protein has a sequence similar to the 40-residue motif of integrases of phages, bacteria and yeast, indicating that this protein could be involved in V-(D)-J recombination as a recombinase. PMID- 2556645 TI - Inhibition of endocytic vesicle fusion in vitro by the cell-cycle control protein kinase cdc2. AB - Membrane transport between the endoplasmic reticulum and the plasma membrane, which involves the budding and fusion of carrier vesicles, is inhibited during mitosis in animal cells. At the same time, the Golgi complex and the nuclear envelope, as well as the endoplasmic reticulum in some cell types, become fragmented. Fragmentation of the Golgi is believed to facilitate its equal partitioning between daughter cells. In fact, it has been postulated that both the inhibition of membrane traffic and Golgi fragmentation during mitosis are due to an inhibition of vesicle fusion, while vesicle budding continues. Although less is known about the endocytic pathway, internalization and receptor recycling are also arrested during mitosis. We have now used a cell-free assay to show that the fusion of endocytic vesicles from baby hamster kidney cells is reduced in Xenopus mitotic cytosol when compared with interphase cytosol. We reconstituted this inhibition in interphase cytosol by adding a preparation enriched in the starfish homologue of the cdc2 protein kinase. Inhibition was greater than or equal to 90% when the added cdc2 activity was in the range estimated for that in mitotic Xenopus eggs, which indicates that during mitosis the cdc2 kinase mediates an inhibition of endocytic vesicle fusion, and possibly other fusion events in membrane traffic. PMID- 2556646 TI - [Chickenpox? But... she could have a baby any moment now!]. PMID- 2556635 TI - Rotavirus gene structure and function. AB - Knowledge of the structure and function of the genes and proteins of the rotaviruses has expanded rapidly. Information obtained in the last 5 years has revealed unexpected and unique molecular properties of rotavirus proteins of general interest to virologists, biochemists, and cell biologists. Rotaviruses share some features of replication with reoviruses, yet antigenic and molecular properties of the outer capsid proteins, VP4 (a protein whose cleavage is required for infectivity, possibly by mediating fusion with the cell membrane) and VP7 (a glycoprotein), show more similarities with those of other viruses such as the orthomyxoviruses, paramyxoviruses, and alphaviruses. Rotavirus morphogenesis is a unique process, during which immature subviral particles bud through the membrane of the endoplasmic reticulum (ER). During this process, transiently enveloped particles form, the outer capsid proteins are assembled onto particles, and mature particles accumulate in the lumen of the ER. Two ER specific viral glycoproteins are involved in virus maturation, and these glycoproteins have been shown to be useful models for studying protein targeting and retention in the ER and for studying mechanisms of virus budding. New ideas and approaches to understanding how each gene functions to replicate and assemble the segmented viral genome have emerged from knowledge of the primary structure of rotavirus genes and their proteins and from knowledge of the properties of domains on individual proteins. Localization of type-specific and cross-reactive neutralizing epitopes on the outer capsid proteins is becoming increasingly useful in dissecting the protective immune response, including evaluation of vaccine trials, with the practical possibility of enhancing the production of new, more effective vaccines. Finally, future analyses with recently characterized immunologic and gene probes and new animal models can be expected to provide a basic understanding of what regulates the primary interactions of these viruses with the gastrointestinal tract and the subsequent responses of infected hosts. PMID- 2556636 TI - Protein phosphorylation and regulation of adaptive responses in bacteria. AB - Bacteria continuously adapt to changes in their environment. Responses are largely controlled by signal transduction systems that contain two central enzymatic components, a protein kinase that uses adenosine triphosphate to phosphorylate itself at a histidine residue and a response regulator that accepts phosphoryl groups from the kinase. This conserved phosphotransfer chemistry is found in a wide range of bacterial species and operates in diverse systems to provide different regulatory outputs. The histidine kinases are frequently membrane receptor proteins that respond to environmental signals and phosphorylate response regulators that control transcription. Four specific regulatory systems are discussed in detail: chemotaxis in response to attractant and repellent stimuli (Che), regulation of gene expression in response to nitrogen deprivation (Ntr), control of the expression of enzymes and transport systems that assimilate phosphorus (Pho), and regulation of outer membrane porin expression in response to osmolarity and other culture conditions (Omp). Several additional systems are also examined, including systems that control complex developmental processes such as sporulation and fruiting-body formation, systems required for virulent infections of plant or animal host tissues, and systems that regulate transport and metabolism. Finally, an attempt is made to understand how cross-talk between parallel phosphotransfer pathways can provide a global regulatory curcuitry. PMID- 2556647 TI - [The role of dietary fiber in gastroenterology: fact or fancy?]. PMID- 2556648 TI - Effects of an N-methyl-D-aspartate receptor agonist and its antagonist CPP on the levels of dopamine and serotonin metabolites in rat striatum collected in vivo by using a brain dialysis technique. AB - 3-((+-)-2-Carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP) is an antagonist at the N-methyl-D-aspartate (NMDA) subtype of glutamate receptor. In the present study, levels of dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and 5-hydroxyindolacetic acid (5-HIAA) were measured after intracerebroventricular injection of NMDA, CPP or both in rat striatum using a brain dialysis method. The injection of NMDA produced a significant increase in DOPAC level. HVA level was also increased by NMDA injection. The level of 5-HIAA was not affected by NMDA injection. The injection of CPP had no effect on DOPAC, HVA and 5-HIAA levels. The injection of CPP restrained the increase of DOPAC and HVA levels induced by NMDA injection. The results suggest that intracerebral injection of NMDA may increase dopamine release from rat striatum, but have no effect on serotonin release. Furthermore, CPP inhibits NMDA induced release of dopamine. PMID- 2556650 TI - Developmental changes of calpain and calpastatin in rabbit brain. AB - A major part of the Ca-activated proteolytic activity in the soluble fraction from rabbit brain could be due to the activity of the neutral thiol-proteases calpain I and II. The activity of calpains exceeded that of the endogenous inhibitor, calpastatin, at all developmental stages studied. The level of calpains increased rapidly from the prenatal stage to reach a peak 10-20 days postnatally. From this period the level of calpains decreased slowly to reach the adult levels. The level of calpastatin increased steadily from the prenatal stage to old age. PMID- 2556649 TI - Early developmental exposure to benzodiazepine ligands alters brain levels of thiobarbituric acid-reactive products in young adult rats. AB - Levels of thiobarbituric acid (TBA)-reactive material were measured in brain regions of 3-4 month-old rats following prenatal exposure to several benzodiazepine (BDZ) receptor ligands over gestational days 14-20. Prenatal exposure to diazepam (DZ) at 1.0 mg/kg/day markedly elevated levels of brain TBA reactive material while exposure to a higher dose (2.5 mg/kg) induced a significant increase only in the hippocampus. Early exposure to the central-type BDZ agonist clonazepam as well as to the central-type antagonist Ro 15-1788 also increased brain levels of TBA-reactive material. Concurrent exposure to the higher dose of DZ partially attenuated the effect of Ro 15-1788. Prenatal exposure to the peripheral-type BDZ ligand PK11195 produced a profound increase in TBA-reactive products in all regions, and concurrent DZ exposure did not attenuate this effect, except in the basal ganglia. Measurement of TBA-reactive material from birth to 3 months indicated that the effect of prenatal exposure to DZ was not apparent until after 8 weeks of age. Acute in vitro exposure of adult and fetal tissue to DZ had no effect on TBA-reactive material. The results suggest an interference in the organization of cellular metabolism in the brain by developmental exposure to BDZ ligands. PMID- 2556651 TI - Lipid peroxidation as the mechanism of modification of the affinity of the Na+, K+-ATPase active sites for ATP, K+, Na+, and strophanthidin in vitro. AB - The effect of lipid peroxidation on the affinity of specific active sites of Na+,K+-ATPase for ATP (substrate), K+ and Na+ (activators), and strophanthidin (a specific inhibitor) was investigated. Brain cell membranes were peroxidized in vitro in the presence of 100 microM ascorbate and 25 microM FeCl2 at 37 degrees C for time intervals from 0-20 min. The level of thiobarbituric acid reactive substances and the activity of Na+, K+-ATPase were determined. The enzyme activity decreased by 80% in the first min. from 42.0 +/- 3.8 to 8.8 +/- 0.9 mumol Pi/mg protein/hr and remained unchanged thereafter. Lipid peroxidation products increased to a steady state level from 0.2 +/- 0.1 to 16.5 +/- 1.5 nmol malonaldehyde/mg protein by 3 min. In peroxidized membranes, the affinity for ATP and strophanthidin was increased (two and seven fold, respectively), whereas affinity for K+ and Na+ was decreased (to one tenth and one seventh of control values, respectively). Changes in the affinity of active sites will affect the phosphorylation and dephosphorylation mechanisms of Na+, K+-ATPase reaction. The increased affinity for ATP favors the phosphorylation of the enzyme at low ATP concentrations whereas, the decreased affinity for K+ will not favor the dephosphorylation of the enzyme-P complex resulting in unavailability of energy for transmembrane transport processes. The results demonstrate that lipid peroxidation alters Na+, K+-ATPase function by modification at specific active sites in a selective manner, rather than through a non-specific destructive process. PMID- 2556652 TI - Role of heme oxygenase in heme-mediated inhibition of rat brain Na+-K+-ATPase: protection by tin-protoporphyrin. AB - Hemoglobin has been shown to inhibit brain Na+-K+-ATPase through an iron dependent mechanism. Both hemoglobin and iron cause spontaneous peroxidation of brain lipids. Release of iron from the heme molecule in animal tissues is dependent on the activity of heme oxygenase. We hypothesized that inhibition of heme catabolism by heme oxygenase prevents the iron-mediated inhibition of Na+-K+ ATPase and might subsequently reduce the tissue damage. Therefore, we studied the effect of heme and tin-protoporphyrin, an inhibitor of heme oxygenase, on the activity of partially purified Na+-K+-ATPase from rat brain in the presence and absence of purified hepatic heme oxygenase. Heme alone at a concentration of 30 microM did not inhibit Na+-K+-ATPase. However, in the presence of heme oxygenase, heme inhibited Na+-K+-ATPase by 75%. Pretreatment of rats with SnCl2, a known inducer of heme oxygenase, reduced the basal activity of the brain Na+-K+-ATPase by 50%. Inhibition of heme oxygenase by tin-protoporphyrin (30 microM) prevented the inhibition of Na+-K+-ATPase which occurred in the presence of heme and heme oxygenase. It is concluded that suppression of heme oxygenase by tin protoporphyrin might be a therapeutic approach to management of hemoglobin associated brain injury following CNS hemorrhage. PMID- 2556653 TI - Differential effects of prazosin and yohimbine on fentanyl-induced muscular rigidity in rats. AB - Whereas muscular rigidity is a well-known phenomenon that is related to anesthesia induced by large doses of narcotic drugs, the precise underlying mechanism(s) remain to be fully elucidated. This study investigated the possible role of noradrenergic neurotransmission and the participation of alpha adrenoceptors in this phenomenon. Male Sprague-Dawley rats, under ketamine induced anesthesia (120 mg/kg, i.p.) and with proper control of respiration, body temperature and end-tidal CO2 were used. Intravenous administration of fentanyl (100 micrograms/kg) consistently caused a significant increase in the electromyographic (EMG) activity, recorded from both gastrocnemius and abdominal rectus muscles. This implied muscular rigidity was markedly antagonized by pretreatment with the specific alpha 1-adrenoceptor blocker, prazosin (50 or 250 micrograms/kg, i.v.). This antagonism occurred in spite of a high level of fentanyl in the plasma, as determined by radioimmunoassay. The specific alpha 2 adrenoceptor blocker, yohimbine (1.15 or 2.3 mg/kg, i.v.), on the other hand, not only failed to prevent fentanyl-induced activation of the EMG, but actually potentiated the response. It is concluded that noradrenergic neurotransmission, possibly originating from the locus coeruleus, may participate in the elicitation of muscular rigidity by fentanyl. Furthermore, this process may involve an excitatory action through alpha 1-, and an inhibitory action through alpha 2 adrenoceptors, in the spinal cord. PMID- 2556654 TI - An electrophysiological study on the common antitussive drug dextromethorphan in hippocampal slices in the rat. AB - Dextromethorphan, a common antitussive drug derived from the opioid class of morphinans, has been reported to have antiepileptic properties in some experimental tests. The effects of dextromethorphan were tested on the synaptic responses of the interconnections between Schaffer collaterals and CA1 pyramidal cells in hippocampal slices of the rat. Dextromethorphan, up to 100-200 microM, did not affect the basal field potentials resulting from electrical stimulation of the radiatum (0.1 Hz, 100-200 microA, 75 mu sec) of CA1 pyramidal cells. Larger concentrations of the drug (300-400 microM) depressed the amplitude and increased the duration of the CA1 basal population spikes. Dextromethorphan, up to 200-300 microM, affected neither the duration nor the incidence of the additional population spikes of the epileptiform bursting, resulting from perfusion of the slice with 1 mM penicillin or in the absence of magnesium ions. The data indicate that: (1) dextromethorphan has an influence on CA1 pyramidal cells, different from that of other opioids; (2) the reported antiepileptic effects of dextromethorphan do not involve the hippocampal area. PMID- 2556655 TI - Selective inhibition by ethanol of glutamate-stimulated cyclic GMP production in primary cultures of cerebellar granule cells. AB - In primary cultures of cerebellar granule cells of the rat, the accumulation of cyclic GMP was stimulated by glutamate, acting at the N-methyl-D-aspartate recognition site, and by atrial natriuretic factor. The response to glutamate was calcium-dependent, while the response to atrial natriuretic factor was not. Ethanol inhibited the accumulation of cyclic GMP in response to both glutamate and atrial natriuretic factor. However, the response to glutamate was much more sensitive to ethanol, with 30-40% inhibition occurring at 50 mM ethanol. Substantial inhibition of the response to atrial natriuretic factor was observed only at concentrations of ethanol of 200 mM or larger. The data suggest that a major site of action of ethanol in inhibiting the accumulation of cyclic GMP is the coupling of the glutamate receptor to soluble guanylate cyclase. The effect of ethanol on agonist-activated activity of guanylate cyclase may contribute to the pharmacological action of ethanol in vivo. PMID- 2556656 TI - Relationship between desensitization and sequestration of muscarinic cholinergic receptors in two neuronal cell lines. AB - Agonist-induced sequestration and desensitization of muscarinic receptors was studied in two cell lines that each express differentially-coupled receptors. The NG108-15 glioma x neuroblastoma hybrid cells have muscarinic receptors coupled only to the inhibition of adenylate cyclase, whereas SK-N-SH human neuroblastoma cells have muscarinic receptors coupled only to the turnover of phosphoinositide. In both NG108-15 and in SK-N-SH cells, muscarinic agonists caused a rapid, temperature-dependent, loss of binding sites for [3H] N-methylscopolamine. In contrast, the density of binding sites for [3H] quinuclidinyl benzilate remained almost unchanged after treatment with carbachol. Since carbachol also caused a redistribution of muscarinic receptors from the plasma membrane to a lighter membrane fraction, the loss of binding sites for [3H] N-methyl-scopolamine was interpreted as being due to sequestration of receptors. In addition to agonist induced sequestration, muscarinic agonists also caused desensitization of the receptor-mediated response in NG108-15 cells. In SK-N-SH cells, however, no such desensitization of the receptor-mediated response was found, despite the agonist induced sequestration of receptors. These findings demonstrate that agonist induced sequestration of receptors does not always lead to desensitization of receptors. PMID- 2556657 TI - Evidence for the involvement of distinct voltage-sensitive calcium channels in the release of 3H-dopamine from primary cultures of mesencephalic neurons. AB - In ventral mesencephalic neurons cultured for five days the K+-evoked 3H-dopamine release is mediated through activation of N-type Ca2+ channels, while L- or T type channels appear to be inactive. In contrast, veratridine-elicited release of 3H-dopamine that was attenuated by tetrodotoxin was not altered by N-, L-, nor T type Ca2+ channel blockers. PMID- 2556658 TI - Gangliosides attenuate NMDA receptor-mediated excitatory amino acid release in cultured cerebellar neurons. AB - Release of both D-[3H]aspartate and endogenous amino acids was measured in primary cultures of cerebellar granule cells. Two hour-pretreatment with the glycosphingolipids, GM1 or GT1b, attenuated the stimulation of excitatory amino acid release induced by depolarizing concentrations of K+ (50 mM). Gangliosides inhibited the phencyclidine (PCP)-sensitive component of depolarization-induced release, i.e. the amplification of release that follows activation of NMDA receptors by the endogenous glutamate. PMID- 2556659 TI - Hippocampal damage induced by ischemia and intra-amygdaloid kainate injection: effect on N-methyl-D-aspartate, N-(1-[2-thienyl]cyclohexyl)piperidine and glycine binding sites. AB - The N-methyl-D-aspartate receptor channel-complex is widely distributed in the hippocampus, particularly in the CA1 region, in the terminal field of CA3 pyramidal axons and in the fascia dentata, in the terminal field of the perforant pathway. In the present study, we have examined, in the rat, the effect of specific lesions of various neuronal populations of the hippocampus on the distribution of several markers of the N-methyl-D-aspartate receptor-channel complex. Anoxic-ischemic treatment produced a destruction of CA1 pyramidal cells (postsynaptic element): this was associated with a 50% loss of N-methyl-D aspartate, glycine and N-(1-phenylcyclohexyl)piperidine binding sites. In contrast, the destruction of CA3 pyramidal cells and their axons (presynaptic element) by kainate treatment did not induce significant changes in the density of binding sites. The present results therefore strongly support an exclusively postsynaptic localization of the N-methyl-D-aspartate receptor-channel complex in CA1; the possibility of a localization of the remaining binding sites on glial cells or interneurons is discussed. In the molecular layer of the fascia dentata, the anoxic-ischemic treatment produced a partial destruction of the median perforant pathway (presynaptic element) associated with a decrease in the density of N-methyl-D-aspartate, N-(1-[2-thienyl]cyclohexyl)piperidine and glycine binding sites; this suggests that, in contrast to CA1, in the molecular layer of the fascia dentata, N-methyl-D-aspartate receptor-binding sites are located both pre- and postsynaptically. PMID- 2556660 TI - Kappa opioid binding sites in the dog cerebral cortex and spinal cord. AB - In the dog cerebral cortex and spinal cord, [3H]bremazocine and [3H]U69593 both bound with high affinity to an apparent single population of binding sites under kappa-selective conditions. In the cortex similar Bmax values for both radioligands in the saturation studies and the high affinity of the kappa selective agents PD117302 and U69593 for both [3H]bremazocine and [3H]U69593 labelled sites in the competition studies suggested a predominance of U69593 sensitive sites previously described as kappa 1 in the guinea-pig and rat brain. The lower slope values for the inhibition curves of PD117302 and U69593 against [3H]bremazocine but not against [3H]U69593 suggested that [3H]bremazocine could also be binding to a relatively minor proportion of additional, possibly kappa 2, sites while [3H]U69593 would appear to be selective for the kappa 1 site. In contrast, in the dog spinal cord, [3H]U69593 appeared to recognize only a proportion (approximately 35%) of the [3H]bremazocine labelled binding site. The significantly lower affinities and slope values of U69593 and PD117302 against [3H]bremazocine were consistent with the additional sites representing the k2 (benzomorphan) sites previously described in guinea-pig and rat spinal cord. Alternatively, the low (micromolar) affinity of the mu-selective ligand, [D-Ala2, MePhe4, Gly-ol5]enkephalin, implied that these additional sites might not be kappa 2 but possibly a low affinity mu site normally expressed under more physiological conditions. PMID- 2556661 TI - Calcitonin gene-related peptide-alpha receptor binding sites in the gastrointestinal tract. AB - Calcitonin gene-related peptide-alpha (CGRP alpha) is a putative neurotransmitter in the brain and in peripheral tissues. Quantitative receptor autoradiography was used to localize and quantify the distribution of specific binding sites for radiolabeled human CGRP alpha in the canine gastrointestinal tract. The canine gastrointestinal tract was chosen as a model since it is similar in both size and structure to the human gastrointestinal tract. In the stomach CGRP alpha binding sites were localized to smooth muscle cells in the muscularis mucosa and muscularis externa, the smooth muscle and endothelium of medium and small arteries, neurons in the myenteric plexus, mucosal epithelial cells and the germinal centers of lymph nodules. In the intestines, the prominent cells types expressing CGRP alpha receptors were myenteric neurons and the germinal centers of lymph nodules. Since previous studies have demonstrated that CGRP-containing sensory neurons innervate the muscularis externa in the stomach and since CGRP alpha receptors are expressed by smooth muscle cells in the muscularis externa, these results suggest that sensory neurons may directly regulate gastric motility by releasing CGRP. In correlation with previous physiological data, the present study suggests that CGRP is involved in the regulation of a variety of gastrointestinal functions including gastric motility, mucosal ion transport, hemodynamics, digestive enzyme secretion, neuronal excitability, and the inflammatory and immune response. PMID- 2556662 TI - Vasoactive intestinal polypeptide receptor binding sites in the human gastrointestinal tract: localization by autoradiography. AB - Vasoactive intestinal polypeptide (VIP) is a putative neurotransmitter in both the brain and peripheral tissues. To define possible target tissues of VIP we have used quantitative receptor autoradiography to localize and quantify the distribution of [125I]VIP receptor binding sites in histologically normal human surgical specimens. While the distribution of VIP binding sites was different for each gastrointestinal segment examined, specific vasoactive intestinal polypeptide binding sites were localized to the mucosa, the muscularis mucosa, the smooth muscle of submucosal arterioles, the circular and longitudinal smooth muscle of the muscularis externa, the myenteric plexus, and lymph nodules. In most segments, the mucosal layer expressed the highest concentration of VIP binding sites, with the duodenal and jejunal mucosa showing the highest density of receptors. These results identify putative VIP target tissues in the human gastrointestinal tract. In correlation with physiological data, VIP binding sites appear to be involved in the regulation of a variety of gastrointestinal functions including mucosal ion transport, gastric secretion, hemodynamic regulation, gastric and intestinal motility, neuronal excitability, and modulation of the immune system. PMID- 2556663 TI - Neurite outgrowth from mouse neuroblastoma and cerebellar cells induced by the protein kinase inhibitor H-7. AB - The protein kinase inhibitor H-7 (1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride) or staurosporine was found to induce neurite outgrowth from mouse neuroblastoma N18TG2 cells and cultured cerebellar cells by light microscopy. Examination by electron microscopy revealed some morphological differences between the neurites of N18TG2 cells induced by H-7 and dB-cAMP, in which H-7 formed longer and thinner neurites and more abundant localized varicose structures along neurites than dB-cAMP. These results indicate that protein kinase inhibitor itself can act as a potent stimulus for inducing neurite outgrowth in a different manner from dB-cAMP. PMID- 2556664 TI - Characterization of neurons containing nerve growth factor receptors in the rat neostriatum. AB - Using a sensitive immunohistochemical procedure, the normal morphology and distribution of neurons containing the nerve growth factor receptors (NGF-R) have been studied in the caudate-putamen of the adult rat brain, and the findings compared with observations on the choline acetyltransferase (ChAT)-positive neurons present in the immediately adjacent sections. The ChAT immunoreaction product was uniformly distributed on the cell bodies, while the NGF-R immunoreaction end product was present also as intensely stained granules (0.06 0.12 micron diameter) on the cell somata and the dendrites. In the NGF-R-positive neurons the intensity of the reaction product was much weaker than the ChAT immunoreactivity. Moreover, in comparison with ChAT-immunopositive cells, the proportion of the NGF-R-immunoreactive neurons was on average about 15% in the neostriatum. PMID- 2556665 TI - Cytochrome oxidase activity in retinas transplanted to the brainstem in rats. AB - Fetal retinas were transplanted to the brainstem of newborn rats and their morphological features were examined using the cytochrome oxidase histochemical method at maturity. The results showed that the inner segments of photoreceptors, outer and inner plexiform layers as well as ganglion cells with large somata were moderately to darkly stained for cytochrome oxidase. This pattern is basically the same as that observed in the normal retina, suggesting that cytochrome oxidase can be used not only for revealing spatial but also functional organization of retinal transplants. PMID- 2556667 TI - Pre-weaning handling reduces adrenocorticotropin response to novel but not to noxious stimuli in adult rats. AB - Effects of handling stimuli given to the rat during pre-weaning period were investigated on plasma immunoreactive (ir) adrenocorticotropin (ACTH) level after electric footshocks or novel audiovisual stimuli in adult life. Plasma ir-ACTH levels after footshocks did not significantly differ between non-handled control and previously handled rats, while the hormone level after novel audiovisual stimuli was significantly lower in handled than in the control rats. These results demonstrate that pre-weaning handling reduces ACTH response to novel audiovisual stimuli but not to footshocks in adult life, and thus suggest the possibility that stress during pre-weaning period affects differentially the developmental plasticity of the hypothalamo-pituitary axis. PMID- 2556666 TI - Ontogeny of strychnine-insensitive [3H]glycine binding sites in rat forebrain. AB - The specific binding sites for strychnine-insensitive [3H]glycine were already demonstrable at prenatal stages and had increased to the adult level by postnatal day (PN) 10. This ontogenic increase was found to be due to an augmentation of Bmax without changes in kd of the [3H]glycine binding. Moreover, there was no shift in inhibition of the binding due to glycine, D-serine, L-serine and HA-966 (1-hydroxy-3-amino-pyrrolid-2-one) after birth. These findings are consistent with the view that there is an increase in density in the absence of the glycine recognition site associated with N-methyl-D-aspartate receptor/ion channel complex in rat forebrain during ontogenic development. PMID- 2556668 TI - Regional distribution of metabotropic glutamate response in the rat brain using Xenopus oocytes. AB - The regional distribution of metabotropic L-glutamate responses was investigated in Xenopus oocytes injected with poly(A)+-RNA from a rat brain which was separated into 3 parts: cerebrum, cerebellum and brainstem. Under voltage-clamp, oscillatory current responses were induced more in cerebellum or brainstem poly (A)+-RNA-injected oocytes, and less in cerebrum poly(A)+-RNA-injected oocytes. These results suggest that the metabotropic glutamate receptor is distributed mostly in cerebellum and brainstem. PMID- 2556669 TI - Lung cancer. Difference in age at diagnosis between men and women. AB - The analysis of a data file comprising 776 lung cancer patients from the Tumor Registry at a 705-bed community hospital in New Jersey reveals that women are diagnosed with squamous cell and large cell lung cancer at an earlier age than men. PMID- 2556670 TI - Control of intracellular calcium in vertebrate photoreceptors. PMID- 2556671 TI - Development of multiple carcinomas in a long term survivor of Wegener's granulomatosis treated with immunosuppressive drugs. PMID- 2556672 TI - Herpes simplex virus. AB - Genital herpes simplex infection remains an infectious disease having widespread consequences for both adult and neonatal populations. Physicians must understand that the psychologic impact of genital HSV infections frequently is more disabling than the physical manifestations. Sensitivity, compassion, and support are necessary from members of the medical community, and psychologic counseling may help patients cope with HSV and all its implications. Similarly, physicians must downplay the hysteria that has been associated with herpesvirus infections, emerging as a result of intense media coverage in the 1970s and 1980s. Minimizing the number of cases of neonatal herpes through identification of infected women during parturition continues to be important. Recent guidelines suggest a rationale for the management of women at risk for genital HSV. Nevertheless, until newer immunologic and serologic techniques become clinically reliable and allow a correct and rapid diagnosis of herpes simplex infection, identification of the infected woman in labor must be made using detailed clinical history, physical examination, and viral culture. PMID- 2556673 TI - Genital papillomavirus infections: current knowledge and future prospects. AB - Although much has been learned about the biology of HPV during the last decade, little progress has been made in the development of rational and effective patient management strategies. Without stronger evidence supporting a causal role for HPV in the development of genital tract and anal cancers, and without the availability of effective therapies, it is premature to recommend widespread screening for detection of HPV DNA or RNA with the molecular hybridization tests soon to be available commercially. Papanicolaou smear screening for detection of early precancerous changes of the cervix and referral for colposcopy and biopsy of areas of epithelium that are suspicious for intraepithelial neoplasia of the cervix, vagina, vulva, penis, or anus remain the cornerstones of genital tract and anal cancer prevention. The patient care implications of subclinical persistent HPV infection of the genital tract are not well understood. For this reason, and because none of the available therapies are curative, treatment of large areas of normal-appearing genital tract epithelium also cannot be recommended at this time. It is hoped that, with the growing research focus on therapies that have the potential for virologic cure, some day effective treatment for subclinical infection will be available. Until that time, patients with recalcitrant or recurring genital warts may benefit most by the sequential application of different treatment modalities. PMID- 2556674 TI - Orofacial herpes simplex virus infections: current concepts in the epidemiology, pathogenesis, and treatment, and disorders in which the virus may be implicated. AB - Herpes simplex virus (HSV) is the causal agent of herpetic stomatitis, recurrent herpes labialis, and the recurrent intraoral infections seen particularly in the immunocompromised patient. HSV has also been implicated in some erythema multiforme, cranial neuropathies, Behcet's syndrome, and oral squamous carcinoma. Recent advances in understanding of HSV, epidemiology, pathogenesis, and treatment are reviewed because there have been numerous advances over the past 5 years. PMID- 2556675 TI - Malignant fibrous histiocytoma arising from the vocal cord. AB - A rare case of malignant fibrous histiocytoma of the left vocal cord occurring in an 80-year-old man is described, with a review of the literature. There has been no recurrence for 24 months since the initial endolaryngeal microsurgery. PMID- 2556676 TI - Effect of CpG-rich sequences in transformation and tumorigenesis by polyomavirus. AB - To address the question of the role of CpG-rich sequences in gene expression, we investigated the effect of an HTF island on the activity of the polyomavirus middle T (pmt) oncogene. pmt is less transformant and less tumorigenic when it is introduced into cultured cells or newborn rats in the presence of an HTF island. Transformed cells carrying pmt in the vicinity of the HTF island have a propensity to revert at a relatively high rate of about 2 X 10(-3) per cell per generation by a mechanism probably involving methylation of some CpG sites in the island. Our results suggest that HTF islands can function as transcriptional silencers. PMID- 2556677 TI - The E6 and E7 genes of HPV-18 are sufficient for inducing two-stage in vitro transformation of human keratinocytes. AB - Using a recently described keratinocyte assay, we demonstrate that HPV-18 DNA induces two progressive steps in cellular transformation (a large cell and a small cell stage). Both steps of this keratinocyte transformation can be achieved with a subgenomic fragment containing only the HPV-18 regulatory region and E6/E7 genes. Similar to cell lines transformed by the complete HPV-18 genome, keratinocytes transformed by the HPV-18 E6/E7 genes express the major early viral protein (E7) but are non-tumorigenic in nude mice. Interestingly, HPV-18 DNA was noted to be 5 times more efficient than HPV-16 DNA for in vitro keratinocyte transformation, regardless of the method of DNA transfection. PMID- 2556678 TI - Extranasopharyngeal angiofibroma. PMID- 2556679 TI - Malignant fibrous histiocytoma of the tongue demonstrated by magnetic resonance imaging. PMID- 2556680 TI - Eimeria tenella, E. acervulina and E. maxima: studies on the development of resistance to diclazuril and other anticoccidial drugs in the chicken. AB - Resistance to diclazuril was induced by 10 passages of the Houghton strains of Eimeria acervulina and E. tenella in chickens given progressively greater concentrations of the drug. This resistance was, however, not complete since the drug retained some efficacy against the drug-passaged lines. Attempts to passage the Houghton strain of E. maxima in birds medicated with concentrations of diclazuril greater than 0.016 parts per million (ppm) were unsuccessful and after 10 passages at this concentration resistance had not developed. Resistance to methyl benzoquate developed after 6 passages of E. tenella in medicated chickens but resistance to amprolium was only partial after 10 passages. The pathogenicity of diclazuril- and amprolium-resistant lines of E. tenella was lower than that of the parental line. Diclazuril was effective against lines of E. tenella resistant to amprolium, arprinocid, clopidol, dinitolmide, halofuginone, methyl benzoquate, monensin and robenidine. PMID- 2556681 TI - Biological effects of lithium chloride on the insect trypanosomatid Herpetomonas samuelpessoai. AB - Effects of lithium chloride on growth, differentiation and respiration of Herpetomonas samuelpessoai, cultivated in a synthetic medium were studied both at 28 and 37 degrees C. Low concentration of lithium chloride (15 mM) stimulated growth at 37 degrees C. In addition, the protozoon tolerated high concentrations (60-150 mM) of the salt at both incubation temperatures. In general, 15 mM lithium chloride increased and 150 mM decreased oxygen uptake when glucose, glutamic acid and proline were used as substrates. However, at 28 degrees C after incubation for 96 h, the highest concentration increased oxygen uptake in the presence of glucose. Sodium butyrate induced cell differentiation in H. samuelpessoai both at 28 and 37 degrees C. High concentration (150 mM) of lithium chloride inhibited cell differentiation of H. samuelpessoai induced by both controlled growth conditions and butyrate addition. The results obtained are described in this paper. PMID- 2556682 TI - [Experimental antioxidant therapy in toxic liver damage from CCl4 and chloxyl]. AB - The authors studied the pharmacotherapeutic efficacy of antioxidants vitamin E, sodium selenite, and their combination in damage to rat liver by CCl4 and the anthelmintic agent chloxyl. Changes of the intensity of peroxidation of biological membrane lipids, the activity of enzymes-markers of hepatocyte cytolysis--alanine aminotransferase and aspartate aminotransferase--in blood serum, and changes in the structure of the liver were studied. Antioxidants and their combination blocked lipid peroxidation, reduced the activity of alanine aminotransferase and aspartate aminotransferase in blood serum considerably, and caused a protective effect on the structure of rat liver in its damage by CCl4 and chloxyl. PMID- 2556683 TI - [New approaches to research on phagocytic cells (a discussion of the article by D. N. Maianskii "The phagocyte system: methodologic problems)]. PMID- 2556684 TI - Uric acid as radical scavenger and antioxidant in the heart. AB - Uric acid (UA) is released from the heart of many species, including man, and its site of formation has been shown to be the microvascular endothelium. Since UA reacts with oxygen radicals in vitro, experiments were conducted on guinea pig hearts perfused with Krebs-Henseleit buffer (KHB) to evaluate whether the formation of UA could afford protection from damage by radicals and oxidants. The following results were obtained: (1) Upon addition of the hydroxyl radical scavenger DMSO to the perfusate, the coronary rate of release of endogenous uric acid was increased relative to the precursor purines. (2) UA was degraded during passage through the coronary system and also in KHB in vitro after addition of substances generating hydroxyl radicals or hypochlorite. Superoxide (O2-) radicals did not seem to react directly with UA, though UA concentration dependently quenched the chemiluminescence generated from luminol in the presence of O2- and OH radicals. (3) Coronary dilation by acetylcholine (Ach) and sub microM concentrations of adenosine, induced by both via endothelial mechanisms, was attenuated after prolonged inhibition of endothelial UA formation by allopurinol. Furthermore, the effect of Ach but not of adenosine proved acutely sensitive to methylene blue and O2-, substances known to inactivate EDRF. This finding suggests involvement of EDRF in Ach-mediated, but not in adenosine induced dilatation of the intact coronary system. Exogenously applied UA prevented the impairment of vascular responses to Ach and adenosine caused by allopurinol, and to Ach upon generation of O2-. (4) Hearts performed more pressure-volume work and exhibited greater functional stability when perfused with KHB supplemented with UA in a physiological concentration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556685 TI - Properties and regulation of chloride channels in cystic fibrosis and normal airway cells. AB - The present study examines the properties of Cl- channels in cultured respiratory cells of cystic fibrosis (CF) patients and normal (N) individuals. In excised membrane patches the conductances for CF and N Cl- channels were larger at positive as compared to negative clamp voltages (Vc): 74 +/- 2.6 (Vc greater than 0) and 47 +/- 2.0 pS (Vc less than 0) for CF (n = 57) and 69 +/- 3.6 (Vc greater than 0) and 45 +/- 2.3 pS (Vc less than 0) for N (n = 35). The open probability (Po) of the channel increased markedly with depolarization. Both the voltage dependence of the conductance and of Po contribute to the outward rectification of the channel. The time histogram analysis reveals two open and two closed time constants. The selectivity of the channel was Cl- = Br- = I- greater than NO-3 much greater than gluconate. The channel was inhibited reversibly by 5-nitro-2-(3 phenylpropylamino)-benzoate (NPPB) at 10(-7) mol/l to 10(-5) mol/l. While Cl- channels were present in cell attached patches of N cells, they were absent in those of CF cells. The mean conductance for cell attached (N) Cl- channels was 76 +/- 3.2 pS for positive clamp voltages (Vc) and 46 +/- 3.9 pS for negative Vc (n = 8). When the membrane patches were excised from CF cells Cl- currents appeared spontaneously (n = 19). The immediate appearance (within 1 s) of Cl- channels after excision was observed at positive (n = 6) as well as at negative clamp voltage (n = 13). "Excision activation" of CF Cl- channels was observed at low (less than 10(-9) mol/l) or high (10(-3) mol/l) calcium activities on the cytosolic side of the excised patch. Variation of the Ca+ activity (less than 10( 9)-10(-3) mol/l) or pH (6.5-8.5) on the cytosolic side exerted no effects on these Cl- channels. These results suggest that Cl- channels are present in the apical membrane of CF and N respiratory cells but they seem to be inhibited in intact CF cells. Excision of the patch and hence removal of the cytosolic "inhibitor" leads to an activation of Cl- channels. The Cl- channels in excised patches of N and CF cells have identical properties. PMID- 2556687 TI - A simple method for multiple fluid exchange. AB - A simple device for fluid exchange is described, which allows the exchange of an unlimited number of solutions at low (10-1000 nl/min) constant perfusion rate. The applicability of the system has been tested in microperfusion experiments of rat distal tubules. At a luminal perfusion rate of 40 nl/min, the lag time was some 20 sec and 80% fluid exchange time some 3 sec. Simple modification allows further reduction of the lag time. Under control conditions, the potential difference across the late distal tubule (PDte) approaches -19.4 +/- 2.5 mV (n = 27). Increase of luminal potassium concentration from 5.4 to 40 mmol/l hyperpolarizes PDte to -29.9 +/- 4.3 mV (n = 8). Amiloride (10 mumol/l) leads to a reversible depolarization to -3.2 +/- 1.0 mV (n = 19), barium (1 mmol/l) to a reversible hyperpolarization to -25.8 +/- 2.6 mV (n = 19). As expected, PDte is largely created by amiloride sensitive sodium channels and is partially blunted by barium sensitive potassium channels. PMID- 2556686 TI - The excretion of highly soluble gases by the lung in man. AB - The excretion (E) of inert gases by the lung depends on, among other things, their blood-gas partition coefficients (lambda). According to conventional gas exchange models, E should increase with increasing lambda. However, recent models that take into account the tidal character of breathing and the buffering capacity of lung tissue predict that E will show a minimum in the range of large lambda values (lambda greater than 10). Further, this local minimum should shift to larger lambda values in exercise conditions as compared to rest conditions. The aim of this study is to verify this predicted behaviour of E. The experiments were carried out with seven healthy subjects at rest and at three work loads (50 W, 100 W and 150 W) on a bicycle ergometer. The behaviour of E was determined from the results of a simultaneous washin of four tracer gases: ethyl acetate (lambda approximately 75), acetone (lambda approximately 330), ethanol (lambda approximately 2000) and acetic acid (lambda approximately 20000). The washin lasted 4 min, and E was calculated from E = 1 - PE-/PI, where PI and PE- are the partial pressures of the tracer gas in inspired and mixed expired gas determined from the recordings obtained during the last minute of washin. PI and PE- were measured with a mass spectrometer. Comparison of the E values of the four gases shows that at rest a minimum value for E is found for acetone. In exercise conditions, however, the smallest E value is found for the more soluble ethanol or acetic acid.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556688 TI - [Influence of hepatic arterial embolization on diabetic patients with hepatocellular carcinoma]. AB - Uncontrollable change of diabetes mellitus (DM) has occurred in one of our patients who had received hepatic arterial embolization (HAE) for hepatocellular carcinoma (HCC). This prompted us to examine the influence of HAE to the diabetic patients with HCC. Thirty-four patients accompanying DM who had received HAE were examined fasted blood glucose (FBG) and the liver function before and after the procedure. HAE was performed using Gelatin Sponge and Lipiodol containing anticancer agents, either alone or combined. Of 34 patients 6 showed increase of FBG level of more than two times after HAE. The FBG level had a tendency to elevate as the grade of DM advanced. The tendency was also recognized on pre-HAE oral glucose tolerance test. However, FBG elevation had no relation to the changes of liver function (GPT, Choline Esterase), the difference of embolic materials and pre-HAE status of DM control. From the results, one must be aware that HAE or Lipiodol infusion to diabetic patients with HCC sometimes may cause uncontrollable change of DM, especially in case of advanced DM patients. Consequently, careful follow-up of HCC as DM is advisable for improvement of the patients' prognosis. PMID- 2556689 TI - Human cytochrome c oxidase during cardiac growth and development. AB - Human heart mitochondrial cytochrome c oxidase specific content and specific activity was measured in five fetuses 15-21 weeks gestational age and in five patients whose age ranged from 6 days to 22 years. None had evidence of cardiac pathology. An increase in cytochrome c oxidase specific content and specific activity was observed in the fetal heart with increasing gestational age (0.13 0.38 nmol heme a/mg protein and 67-295 nmol O2 utilized/min/mg protein) and from the neonatal period (0.35 nmol heme a/mg protein and 140 nmol O2/min/mg protein) to adulthood (1.2 nmol heme a/mg protein and 1104 nmol O2/min/mg protein). A marked increase was observed postnatally between 4 and 19 months. PMID- 2556690 TI - Hepatoma with right atrial extension. AB - The case is described of a 14-year-old boy who had a hepatoma with a right atrial extension. He presented with edema, abdominal pain, and ascites. Two-dimensional echocardiography showed a right atrial tumor that had invaded from the inferior vena cava as an extension into the right atrium of the hepatoma. PMID- 2556691 TI - Antiserotonin activity of 9-methyl-2-[3-(4-phenyl-1-piperazinylpropyl)]-1,2,3,4 tetrahydro -beta-carbolin-1-one (B-193). AB - The effect of B-193 on the central and peripheral serotonin system was studied. B 193 antagonized the head-twitches responses induced by L-5-hydroxytryptophan (L-5 HTP) in mice (ED50 = 0.75 mg/kg ip and 6.6 mg/kg po) and lysergic acid diethylamide (LSD) in rats (ED50 = 1.54 mg/kg ip) and also counteracted forepaws clonic convulsions induced by tryptamine (ED50 = 3.07 mg/kg ip). B-193 (2.5-20 mg/kg ip) antagonized dose-dependently hyperthermia induced by fenfluramine or m chlorophenylpiperazine (m-CPP) and in a dose of 1 mg/kg iv abolished the stimulation of the flexor reflex evoked by quipazine or fenfluramine. B-193 given in a concentration of 10(-7)-10(-5) mol/l competitively inhibited contractions of the rat stomach fundus strip induced by serotonin (5-HT) (pA2 = 6.5) and the increases in blood pressure induced by 5-HT in pithed rats (ED50 = 0.17 mg/kg iv). In receptor binding studies B-193 has shown distinct affinity to 5-HT2 receptors, and alpha 1-adrenoceptors much weaker affinity to 5-HT1 receptors and alpha 2-adrenoceptors but not to beta-adrenergic, GABA-ergic or benzodiazepine receptors. Our findings demonstrated that B-193 shows potent central and peripheral antiserotonin activity. PMID- 2556692 TI - Separation of mature rat ventral prostate epithelial and fibroblast cells. AB - Several techniques for the separation of rat ventral prostate cells, using density gradient centrifugation or mechanical means have been published, yielding fibroblast and epithelial cell populations of varying purities and viability. These techniques are often tedious, yield relatively limited numbers of cells, demand considerable technical expertise, and result in the isolation of cells of limited viability. Two techniques for the isolation and establishment of epithelial and fibroblast cell cultures from mature rat ventral prostate are described here. Collagenase/trypsin digestion of the tissue yields a single-cell suspension of both cell types that are separated on Percoll isopycnic centrifugation gradients. A continuous gradient system allows for the separation of a greater number of cells with very high degrees of purity. A second technique based on a step-gradient system produces reproducible subfractionation of the epithelial cell component of the prostate within a considerable shorter period of time. An improved medium for plating epithelial and fibroblast cells has also been developed. The separated cells are plated on collagen and/or fibronectin coated dishes in a serum-free plating medium that is later replaced with a serum containing growth medium. The plating medium greatly increases the plating efficiency of the isolated cell types, particularly the epithelial cells. PMID- 2556693 TI - Artefacts associated with a cryoglobulin. AB - A 70 year old man is described in whom the presence of a cryoglobulin caused a number of unusual laboratory features which were not recognized as being due to a cryoglobulin initially. Pseudoleucocytosis has been recognized in the past. However, the intracytoplasmic inclusion bodies have only rarely been described. The curious appearance of the ESR has not previously been described. PMID- 2556694 TI - Cholinomimetic drugs modify betamethasone suppression in the rat. AB - This study was performed to investigate the effect of different cholinomimetic drugs (CD) on ACTH levels in betamethasone (BET) pretreated rats (40 mg/kg in the drinking water for 24 h). Animals were injected with atropine methylbromide (1 mg/kg, i.p.) and, after 10 min, with physostigmine (25, 50, 100, 200 micrograms/kg), arecoline (AR) or oxotremorine (OX) (150, 300 micrograms/kg) intravenously. They were killed 10 and 20 min after CD injection, to measure ACTH plasma levels. The lowest dose of AR and both doses of OX were able to induce an escape from BET suppression. The hypothesis that a stimulation of central muscarinic (and, probably, not nicotinic) receptors is involved in the CD-induced escape from glucocorticoid suppression is discussed. PMID- 2556695 TI - [Microglandular pancreatic cancer. Histological, immunocytochemical and electron microscopy findings]. PMID- 2556696 TI - [Mechanisms of latency of the herpes simplex virus]. PMID- 2556697 TI - Tissue-type plasminogen activator variants with domain duplications and rearrangements. AB - The interactions between tPA domains that are important for catalysis are poorly understood. We have probed the function of interdomain interactions by generating tPA variants in which domains are duplicated or rearranged. The proteins were expressed in a transient mammalian expression system and tested in vitro for their ability to activate plasminogen, induce fibrinolysis and bind to a forming fibrin clot. Duplication of the heavy chain domains of tPA produced enzymatically active tPA variants, many of which demonstrated similar in vitro amidolytic and fibrinolytic activity and similar fibrin affinity to the parent molecule. Zymographic analysis of the domain duplication tPA variants showed one major active species for each variant. Selection of the residues duplicated and the interdomain spacing were found to be critical considerations in the design of tPA variants with duplicated domains. We also rearranged the domains of tPA such that kringle 1 replaced the second kringle domain and vice versa. An analysis of these variants indicates that the first kringle domain can confer fibrin affinity to a tPA variant and function in place of kringle 2. Therefore, in wild-type tPA, the functions of kringle 1 and kringle 2 must be dependent partially on their orientation within the heavy chain of the protein. The functional autonomy of the heavy and light chains of tPA is demonstrated by the activity of a tPA variant in which the order of the heavy and light chains was reversed. PMID- 2556698 TI - A versatile and potentially general approach to the targeting of specific cell types by retroviruses: application to the infection of human cells by means of major histocompatibility complex class I and class II antigens by mouse ecotropic murine leukemia virus-derived viruses. AB - A technique for delivering genes carried by recombinant retroviruses into specific cell types could have numerous applications in oncology, developmental biology, and gene therapy. As a first step toward this remote goal we designed a procedure allowing in vitro cell targeting by retroviruses. Biotinylated antibodies against the viral envelope protein on one side, and against specific cell membrane markers on the other side, were bridged by streptavidin and used to link the virus to the host. The method was successfully used to infect human cells with ecotropic murine retroviruses by means of major histocompatibility complex class I and class II antigens and appears easily adaptable to other cell membrane markers. Moreover, the sequential protocol we designed, although allowing infection of human cells, requires less stringent safety constraints than would handling of amphotropic virus stocks. PMID- 2556699 TI - A retinoic acid-responsive element is present in the 5' flanking region of the laminin B1 gene. AB - The retinoic acid (RA)-associated differentiation of murine F9 teratocarcinoma stem cells results in dramatic changes in gene expression. The cellular gene encoding the B1 subunit of the extracellular matrix protein laminin is transcriptionally activated by RA, and its transcription is further enhanced by N6,O2'-dibutyryladenosine 3',5'-cyclic monophosphate (Bt2cAMP) during the differentiation of F9 stem cells into extraembryonic parietal endoderm cells. We now report that expression vectors encoding the human RA receptors RAR-alpha, RAR beta, and RAR-gamma can activate chloramphenicol acetyltransferase (CAT) expression from laminin B1 promoter/CAT expression vectors (e.g., p1.6LAMCAT) in RA-treated F9 cells, as measured in a transient transfection assay. Bt2cAMP does not further enhance the RA-associated increase in CAT activity. Through the use of deletion and mutation analyses, the RA-responsive element (RARE) of the murine laminin B1 gene has been defined as a 46-base-pair element between -477 and -432 of the laminin B1 5' flanking region. Insertion of a region of DNA containing this RARE in either orientation into a thymidine kinase promoter/CAT expression vector causes CAT expression to be activated 5- to 9-fold by the cotransfected human RAR-alpha or RAR-beta constructs in RA-treated F9 cells, and this RARE also functions in human HeLa cells. In contrast, this RARE in the p1.6LAMCAT vector does not activate CAT expression when cotransfected into F9 stem cells with the c erbA gene in the presence of thyroid hormone. This suggests that the laminin B1 gene is activated by RA but not by thyroid hormone in vivo. PMID- 2556700 TI - Escherichia coli replication termination protein impedes the action of helicases. AB - Identification of the consensus sequence for termination of replication (ter) in Escherichia coli and the isolation of the ter-binding protein (TBP) allowed us to test their effects on replication forks initiated at the unique origin of the E. coli chromosome (oriC) in a purified enzyme system. Replication was severely impeded by ter in a unique orientation when purified TBP was supplied to bind it. The target for blockage within the replication complex can now be ascribed to the inability of dnaB helicase to separate the duplex strands when it encounters ter bound by TBP. Other helicases, such as rep and uvrD proteins, that translocate on DNA and displace strands in the direction opposite to that of dnaB protein are also blocked, but only when the TBP-bound ter is oriented in the other direction. From these results, we infer that the orientation of ter confers a particular polarity on the TBP seated on it, such that a helicase is blocked when it confronts TBP from one side, but can act, presumably by displacing TBP, when facing its other side. Thus, the intrinsic nature of the oriented TBP-ter complex is responsible for impeding the helicases, rather than any protein-protein interactions. PMID- 2556701 TI - Recombinant baculoviruses as vectors for identifying proteins encoded by intron containing members of complex multigene families. AB - Using a transfer vector derived from Bombyx mori nuclear polyhedrosis virus (BmNPV), we have constructed recombinant baculoviruses that contain complete silk moth chorion chromosomal genes encoding high-cysteine proteins under the control of the polyhedrin promoter. Silk moth tissue culture cells infected with these recombinant viruses were found to contain abundant RNA sequences of sizes similar to those of the authentic chorion mRNAs. Chorion transcripts present in infected cells were initiated almost exclusively at the cap site of the polyhedrin start site. Primer extension and RNase protection experiments revealed that a considerable proportion of the resultant transcripts were spliced at the same sites as those utilized in follicular cells for the production of functional chorion mRNA. Electrophoretic analysis and immunoprecipitation of the proteins of host cells infected with the recombinant viruses revealed the presence of the corresponding chorion proteins. We conclude that baculovirus vectors can be used for expressing efficiently not only cDNAs or simple genes devoid of intervening sequences but also intron-containing chromosomal genes. Thus, recombinant baculoviruses offer a powerful alternative to hybrid-selected translation, particularly when the identification of proteins encoded by members of complex multigene families is required. PMID- 2556702 TI - Specific gene suppression by engineered ribozymes in monkey cells. AB - Short catalytic RNAs possessing specific endoribonuclease activity (ribozymes) have recently been designed that can potentially shear any chosen target RNA in trans at a specific site. Here, engineered ribozymes targeted against chloramphenicol acetyltransferase (CAT), derived from Tn9, have been cloned into a mammalian expression vector and tested in transient transfection experiments for their effects on CAT expression in monkey (COS1) cells. The ribozymes contained the catalytic domain of the satellite RNA from tobacco ringspot virus and were targeted to three sites in the CAT mRNA by flanking antisense sequences. These ribozymes, which were previously shown to accurately cleave CAT message in vitro, were cloned into a replicating plasmid vector under the control of the highly active simian virus 40 early promoter. The ribozyme gene sequence was incorporated into the 3' untranslated region of the gene for firefly luciferase as it was ineffective when expressed as a short RNA. Each ribozyme construction gave a similar level of suppression of CAT activity when the target was transcribed from the herpes virus thymidine kinase promoter. One of the three (ribozyme 2) was chosen for further study and tested after it had been modified by the addition of extra flanking bases. The reporter gene for luciferase was used to monitor ribozyme level and to function as a specificity control, and the human growth hormone gene was cotransfected as an independent reporter for specificity of the ribozyme against the intended target CAT. At high (approximately 1000-fold) molar excess this ribozyme was demonstrated to consistently and specifically suppress CAT expression (up to approximately 60%) in COS1 cells relative both to a plasmid clone with the ribozyme inserted in the reversed (inactive) orientation and to a control corresponding to the relevant 26 nucleotide antisense segment of CAT. PMID- 2556703 TI - Covalently bound VirD2 protein of Agrobacterium tumefaciens protects the T-DNA from exonucleolytic degradation. AB - We show that upon induction of Agrobacterium tumefaciens, free linear double stranded T-DNA molecules as well as the previously described T-strands are generated from the Ti plasmid. A majority of these molecules are bound to a protein. We show that this protein is the product of the virulence gene virD2. This protein was found to be attached to the 5' terminus of processed T-DNA at the right border and to the rest of the Ti plasmid at the left border. The protein remnant after Pronase digestion rendered the right end of the double stranded T-DNA resistant to 5'----3' exonucleolytic attack in vitro. The protein DNA association was resistant to SDS, mercaptoethanol, mild alkali, piperidine, and hydroxylamine, indicating that it involves a covalent linkage. The possible involvement of this T-DNA-protein complex in replication, transduction to the plant, nuclear targeting, and integration into the plant nuclear DNA is discussed. PMID- 2556705 TI - ExoMeth sequencing of DNA: eliminating the need for subcloning and oligonucleotide primers. AB - A method is reported for sequencing DNA based on exonuclease III digestion and strand protection by using modified nucleoside triphosphates. Up to 10 kilobases of sequence information may be obtained from each strand of a given template without subcloning. Prior knowledge of the restriction map is not important; prior knowledge of any of the sequence is not required. Nor are oligonucleotide primers needed. Double-stranded cosmids, plasmids, lambda phage, or linear molecules (including amplified molecules) may be used as starting material. The method creates a single-stranded template from these starting molecules, thus generating high-quality sequence ladders. Most commonly used DNA polymerases may be utilized, including reverse transcriptase and T7 DNA polymerase. The approach is "ordered", so little time is wasted on redundant sequencing. PMID- 2556704 TI - Cloning of human calcineurin A: evidence for two isozymes and identification of a polyproline structural domain. AB - Two types (I and II) of cDNAs encoding the large (A) subunit of calcineurin, a calmodulin-regulated protein phosphatase, were isolated from human basal ganglia and brainstem mRNA. The complete sequences of the two calcineurin clones are identical except for a 54-base-pair insert in the type I clone and different 3' ends including part of the coding sequence for the C termini of the two proteins. These findings suggest that calcineurin A consists of at least two isozymes that may result from alternative splicing events. The two forms of the enzyme differ in the C terminus, which contains an inhibitory domain rapidly severed by limited proteolysis. With the exception of an 18-amino acid insert, the central parts of the molecules, which harbor the catalytic domains, are identical and show extended similarities with the entire catalytic subunits of protein phosphatases 1 and 2A, defining a distinct family of protein phosphatases. The 40-residue N terminal fragment, specific for calcineurin, contains a sequence of 11 successive prolines that is also found in bovine brain calcineurin by peptide sequencing. A role in the calmodulin activation of calcineurin is proposed for this novel structural element. PMID- 2556706 TI - Aspartic acid-96 is the internal proton donor in the reprotonation of the Schiff base of bacteriorhodopsin. AB - Above pH 8 the decay of the photocycle intermediate M of bacteriorhodopsin splits into two components: the usual millisecond pH-independent component and an additional slower component with a rate constant proportional to the molar concentration of H+, [H+]. In parallel, the charge translocation signal associated with the reprotonation of the Schiff base develops a similar slow component. These observations are explained by a two-step reprotonation mechanism. An internal donor first reprotonates the Schiff base in the decay of M to N and is then reprotonated from the cytoplasm in the N----O transition. The decay rate of N is proportional to [H+]. By postulating a back reaction from N to M, the M decay splits up into two components, with the slower one having the same pH dependence as the decay of N. Photocycle, photovoltage, and pH-indicator experiments with mutants in which aspartic acid-96 is replaced by asparagine or alanine, which we call D96N and D96A, suggest that Asp-96 is the internal proton donor involved in the re-uptake pathway. In both mutants the stoichiometry of proton pumping is the same as in wild type. However, the M decay is monophasic, with the logarithm of the decay time [log (tau)] linearly dependent on pH, suggesting that the internal donor is absent and that the Schiff base is directly reprotonated from the cytoplasm. Like H+, azide increases the M decay rate in D96N. The rate constant is proportional to the azide concentration and can become greater than 100 times greater than in wild type. Thus, azide functions as a mobile proton donor directly reprotonating the Schiff base in a bimolecular reaction. Both the proton and azide effects, which are absent in wild type, indicate that the internal donor is removed and that the reprotonation pathway is different from wild type in these mutants. PMID- 2556707 TI - The membrane binding domain of rod cGMP phosphodiesterase is posttranslationally modified by methyl esterification at a C-terminal cysteine. AB - Retinal rod cGMP phosphodiesterase (3',5'-cyclic-GMP phosphodiesterase; EC 3.1.4.35; PDE), a key regulatory enzyme involved in visual excitation, is one of several outer segment membrane proteins that are carboxyl methylated in the presence of the methyl donor S-adenosyl-L-[3H-methyl]methionine. By chromatographic analyses of the 3H-methyl amino acid generated by exhaustive proteolysis of purified PDE, followed by performic acid oxidation of the digest, we have shown that this modification occurs at a C-terminal cysteine residue of the alpha subunit of this enzyme. When PDE is subjected to limited proteolysis with trypsin, a 3H-methylated fragment of 1000 daltons or less is rapidly removed prior to the degradation of its inhibitory gamma subunit. This small fragment remains membrane bound, whereas the bulk of the enzyme is released, indicating that a domain responsible for anchoring PDE to the membrane is located near the C terminus. Based on the C-terminal amino acid sequence of Cys-Cys-Val-Gln predicted from the alpha cDNA sequence, we conclude that PDE undergoes posttranslational modifications, including the proteolytic removal of two or three terminal amino acids, and methyl esterification of the alpha-carboxyl group of the terminal cysteine residue. We speculate that the sulfhydryl group of the methylated cysteine is also lipidated to mediate membrane binding. These modifications may play an important role in delivering the nascent PDE chains to the membrane and in correctly positioning the PDE molecule in the rod disks for phototransduction. PMID- 2556708 TI - Interaction of a nuclear location signal with isolated nuclear envelopes and identification of signal-binding proteins by photoaffinity labeling. AB - The nuclear envelope (NE) separates the two major compartments of eukaryotic cells, the nucleus and the cytoplasm. Recent studies suggest that the uptake of nuclear proteins into the nucleus is initiated by binding of nuclear location signals (NLSs) contained within these proteins to receptors in the NE, followed by translocation through the nuclear pore complex. To examine the binding step without interference from intranuclear events, we have used a system consisting of (i) purified rat liver NEs fixed onto glass slides and (ii) the prototype simian virus 40 large T antigen (SV40 T) NLS conjugated to nonnuclear carrier proteins, and we have visualized the receptor-ligand interaction by indirect immunofluorescence. In this system, incubation of isolated NEs with the wild-type SV40 T NLS conjugate with carrier proteins resulted in binding that was signal sequence-dependent, could be competitively blocked with excess conjugated and unconjugated wild-type peptide, did not require ATP, and was not affected by the transport-inhibiting lectin wheat germ agglutinin. In contrast, only minimal binding was observed with a mutant SV40 T NLS conjugate. These results are consistent with those obtained in other, more complex in vitro systems and suggest that binding of the SV40 T NLS is receptor-mediated. Binding is largely abolished by extraction of the NE with the nonionic detergent Triton X-100, suggesting that the receptor is soluble in detergent. We find in the Triton X-100 supernatant four major NLS-binding proteins with apparent molecular masses of 76, 67, 59, and 58 kDa by photoaffinity labeling with a highly specific crosslinker, azido-NLS. The reduced complexity of the system described here should be useful for the functional study of other potential NLSs for the identification and isolation of their binding sites and for the screening of antibodies raised against these binding sites. PMID- 2556710 TI - Sequences near the termini are required for transposition of the maize transposon Ac in transgenic tobacco plants. AB - Deletion derivatives of the maize transposable element Activator (Ac) were constructed in vitro and inserted into a kanamycin resistance gene. These constructions were then introduced into tobacco protoplasts derived from plants previously transformed with Ac. The ability of each deletion derivative to excise was measured by whether or not kanamycin-resistant tobacco calli were recovered. This allowed us to determine the length of DNA present at each terminus that is required to respond to the products expressed by the Ac element present in the genome. We show that around 200 base pairs (bp) are required at both ends for excision to occur at wild-type levels. When between 100 and 200 bp were retained at one of the ends, reduced frequencies of excision were detected. With less than 100 bp remaining at either end, no excision was detected. In addition, we show that although similar lengths of DNA are required at each terminus, the termini are not interchangeable. The significance of these data is discussed with respect to the protein(s) which interact(s) with the termini of Ac. PMID- 2556709 TI - Inositol trisphosphate and diacylglycerol can differentially modulate gene expression in Dictyostelium. AB - We have previously shown that several genes expressed during Dictyostelium development could be induced in shaking culture by exogenous cAMP, even though the accumulation of intracellular cAMP was inhibited. The use of selected cAMP analogs indicated that the exogenous cAMP functioned by activating the cell surface cAMP receptor and not by interacting with the regulatory subunit of the intracellular cAMP-dependent protein kinase. Although some genes in Dictyostelium appear to be regulated by intracellular cAMP, these data suggest that this is not the case for all genes regulated by cAMP. Intracellular second messengers other than cAMP may, therefore, promote the expression of these other genes. Here, we have examined inositol trisphosphate and diacylglycerol as candidates for such mediators of signal transduction. We have studied three genes that exhibit disparate modes of temporal and spatial expression during development of Dictyostelium. In shaking cultures, maximal levels of expression of each are dependent on the accumulation of or exposure to extracellular cAMP. We show that the addition of inositol trisphosphate and/or diacylglycerol to cells in shaking culture has distinct effects on the expression of each gene and, under specific conditions, can bypass the requirement for extracellular cAMP. These data suggest that extracellular cAMP interacting with its cell surface receptor may promote synthesis of inositol trisphosphate and diacylglycerol to regulate gene expression and aspects of differentiation in Dictyostelium. PMID- 2556711 TI - Targeted gene mutations in Drosophila. AB - A cloned gene can be of interest because of its expression in a particular tissue or at a certain developmental stage, or because of homology to an interesting gene from another organism. In Drosophila its location in the genome is readily determined by in situ hybridization to the banded larval salivary gland polytene chromosomes, but it is more difficult to isolate mutations that may reveal its function. This paper describes a general method for detecting transposable element insertions into the gene in question. This "reverse genetics" then offers the possibility of observing a consequent mutant phenotype, providing a key to the normal function of the gene. The sensitivity of the polymerase chain reaction makes it possible to detect the occurrence of a single appropriate P-element transposon insertion among a population of mutagenized flies. This is accomplished by the use of oligonucleotide primers--one a sequence from within the cloned gene and the other homologous to the terminal sequence of the P element DNA--to prime synthesis into the DNA flanking an insertion site. A segment of DNA, bounded by the two primers, will be a target for amplification only in a fly in which a P-element has inserted within about 2 kilobases of the gene primer. This technique has been used to detect P-element insertions near a gene expressed in the Drosophila compound eye. Potential problems with the technique and possible refinements in the screen are discussed. In principle, it could be utilized to detect insertion of a foreign element into any gene for which at least a partial sequence is known and could be extended to other organisms. PMID- 2556713 TI - Chromosome-breaking structure in maize involving a fractured Ac element. AB - Chromosome breakage in maize can result from an interaction between certain transposable elements. When an Ac (Activator) element and a state I Ds (Dissociation) element are present together in the genome, either linked or unlinked, breaks occur regularly at the locus of the Ds element. We show here that breaks occur with high frequency at or near the locus of a structure consisting of a 2.5-kilobase (kb) terminally deleted or fractured Ac element very tightly linked to a second, intact 4.6-kb Ac element. This structure has the features of a macrotransposon and may behave like one. Loss of the tight linkage abolishes chromosome breakage. A model based on transposition of the macrotransposon is proposed to explain the chromosome-breaking properties of Ac and Ds. PMID- 2556712 TI - Characterization and immunological identification of cDNA clones encoding two human DNA topoisomerase II isozymes. AB - Several DNA topoisomerase II (Topo II; EC 5.99.1.3) partial cDNA clones obtained from a human Raji-HN2 cDNA library were sequenced and two classes of nucleotide sequences were found. One member of the first class, SP1, was identical to an internal fragment of human HeLa cell Topo II cDNA described earlier. A member of the second class, SP11, shared extensive nucleotide (75%) and predicted peptide (92%) sequence similarities with the first two-thirds of HeLa Topo II. Each class of cDNAs hybridized to unique, nonoverlapping restriction enzyme fragments of genomic DNA from several human cell lines. Synthetic 24-mer oligonucleotide probes specific for each cDNA class hybridized to 6.5-kilobase mRNAs; furthermore, hybridization of probe specific for one class was not blocked by probe specific for the other. Antibodies raised against a synthetic SP1-encoded dodecapeptide specifically recognized the 170-kDa form of Topo II, while antibodies raised against the corresponding SP11-encoded dodecapeptide, or a second unique SP11-encoded tridecapeptide, selectively recognized the 180-kDa form of Topo II. These data provide genetic and immunochemical evidence for two Topo II isozymes. PMID- 2556714 TI - Lithium chloride potentiates tumor necrosis factor-mediated cytotoxicity in vitro and in vivo. AB - Tumor necrosis factor (TNF) is cytotoxic for several transformed cell lines in vitro. In the presence of LiCl, the murine fibrosarcoma cell lines L929 and WEHI 164 clone 13 became greater than 10 times more sensitive to TNF-mediated cytotoxicity. The human tumor cell lines BT20 and HeLa D98/AH2 were also responsive to the cytotoxicity-enhancing effect of LiCl. Other monovalent or divalent cations did not affect TNF-mediated cytotoxicity. The potentiating effect of LiCl on TNF cytotoxicity was largely independent of transcription, and LiCl could be added to the cells as early as 2 hr before or as late as 4 hr after TNF without loss of effectiveness. The mechanism by which LiCl increases the cytotoxic response seems to differ from the sensitizing effect of actinomycin D or interferon gamma, since the latter treatments overcame TNF resistance of several cell lines, whereas LiCl did not. Evidence is presented that LiCl acts, either directly or indirectly, via the TNF-activated phospholipase A2 pathway. In nude mice, a combination of TNF and LiCl led to hemorrhagic necrosis and growth inhibition of L929 tumors, whereas little effect was observed when TNF was administered alone. HeLa D98/AH2 tumors also were sensitive to the potentiating effect of LiCl in vivo. We conclude that LiCl enhances the effectiveness of TNF in vitro and in vivo, results that may have therapeutic implications. PMID- 2556715 TI - Detection of "deleted" mitochondrial genomes in cytochrome-c oxidase-deficient muscle fibers of a patient with Kearns-Sayre syndrome. AB - Using in situ hybridization and immunocytochemistry, we studied a muscle biopsy sample from a patient with Kearns-Sayre syndrome (KSS) who had a deletion of mitochondrial DNA (mtDNA) and partial deficiency of cytochrome-c oxidase (COX; EC 1.9.3.1). We sought a relationship between COX deficiency and abnormalities of mtDNA at the single-fiber level. COX deficiency clearly correlated with a decrease of normal mtDNA and, conversely, deleted mtDNA was more abundant in COX deficient fibers, especially ragged-red fibers. The distribution of mtRNA had a similar pattern, suggesting that deleted mtDNA is transcribed. Immunocytochemistry showed that the nuclear DNA-encoded subunit IV of COX was present but that the mtDNA-encoded subunit II was markedly diminished in COX deficient ragged-red fibers. Because the mtDNA deletion in this patient did not comprise the gene encoding COX subunit II, COX deficiency may have resulted from lack of translation of mtRNA encoding all three mtDNA-encoded subunits of COX. PMID- 2556716 TI - Two epithelial tumor cell lines (HNE-1 and HONE-1) latently infected with Epstein Barr virus that were derived from nasopharyngeal carcinomas. AB - Two epithelial tumor cell lines were established from biopsy specimens of nasopharyngeal carcinomas (NPC). The specimens were taken from poorly differentiated squamous cell carcinomas of the nasopharynx. The tissues were prepared for cell culture and eventually two continuous epithelial cell lines were obtained and designated HONE-1 and HNE-1. Light and electron microscopic examination of these two cell lines demonstrated cells with an epithelial morphology including the presence of desmosomes. The HNE-1 cell line has been passaged more than 100 times and the HONE-1 cell line has been passaged more than 90 times. It was found that early-passage uncloned HNE-1 cells (passage 23) could be superinfected with the B95-8 and NPC-EBV isolates as demonstrated by the induction of Epstein-Barr virus (EBV)-specific early antigen(s) in a small percentage of the cells; HONE-1 cells could also be superinfected with EBV. Southern blot analysis detected EBV DNA in samples from uncloned HNE-1 cells at passages 12, 17, 21, 27, and 35. However, by passage 45, EBV DNA could no longer be detected in HNE-1 cells by Southern blot analysis. The EBV genome was detected in parental HONE-1 cells at subculture 9 and in clone 40 cells up to passage 40 thus far. When HNE-1 cells were examined for the expression of the EBV-encoded nuclear antigen (EBNA) at passage 12, only about 10% of the cells were found to be positive. The percentage of EBNA-positive HNE-1 cells decreased as the cells were passaged. A similar loss of EBNA was observed in uncloned HONE-1 cells, but not in HONE-1 clone 40 cells. In clone 40, which has been passaged 40 times thus far, 85-90% of the cells are still EBNA-positive. The data suggest that EBV genome-positive HNE-1 and HONE-1 cells were lost as the cells were cultivated in vitro and that cloning the cells at an early passage level may be critical in maintaining EBV genome-positive epithelial NPC cells. These EBV genome-positive epithelial NPC cell lines will be useful for studying the association of EBV and NPC. PMID- 2556717 TI - Epstein-Barr virus nuclear protein 2 is a key determinant of lymphocyte transformation. AB - Epstein-Barr virus (EBV) efficiently transforms B lymphocytes to perpetual proliferation. The EBV laboratory strain P3HR-1 is transformation-incompetent and lacks a DNA segment that includes the EBV nuclear antigen 2 (EBNA-2) gene and a portion of the EBNA leader protein (EBNA-LP) gene. These two genes are expressed in transformed B lymphocytes. Recombinant transformation-competent EBVs were produced by transfecting P3HR-1-infected cells with a cosmid containing the DNA deleted in P3HR-1. Deletion of 105 nucleotides from the middle of the EBNA-2 gene had no discernible affect on transformation. Two larger EBNA-2 deletions abolished transformation but did not affect EBNA-2 nuclear localization. Two naturally occurring EBV variants (EBV types 1 and 2) differ extensively in their growth-transformation phenotype and in their EBNA-LP, EBNA-2, and EBNA-3A, -3B, and -3C genes. Recombinant P3HR-1 carrying EBV-1 EBNA-2 has many of the EBV-1 in vitro growth-transforming effects; recombinant P3HR-1, isogenic except for EBV-2 EBNA-2, has many of the EBV-2 growth-transforming effects including slow emergence of transformants, growth in tight clumps with few surrounding viable cells, and early sensitivity to dilution with fresh medium. Thus, EBNA-2 is an essential molecule in lymphocyte growth transformation by EBV and a major determinant of the differences between EBV-1 and EBV-2 in lymphocyte growth transformation. PMID- 2556718 TI - A mechanism for the Hebb and the anti-Hebb processes underlying learning and memory. AB - In a previous paper, a model was presented showing how the group of Ca2+/calmodulin-dependent protein kinase II molecules contained within a postsynaptic density could stably store a graded synaptic weight. This paper completes the model by showing how bidirectional control of synaptic weight could be achieved. It is proposed that the quantitative level of the activity-dependent rise in postsynaptic Ca2+ determines whether the synaptic weight will increase or decrease. It is further proposed that reduction of synaptic weight is governed by protein phosphatase 1, an enzyme indirectly controlled by Ca2+ through reactions involving phosphatase inhibitor 1, cAMP-dependent protein kinase, calcineurin, and adenylate cyclase. Modeling of this biochemical system shows that it can function as an analog computer that can store a synaptic weight and modify it in accord with the Hebb and anti-Hebb learning rules. PMID- 2556719 TI - Nonpsychotropic cannabinoid acts as a functional N-methyl-D-aspartate receptor blocker. AB - Binding studies using the enantiomers of the synthetic cannabinoid 7-hydroxy delta 6-tetrahydrocannabinol 1,1-dimethylheptyl homolog in preparations of rat brain cortical membranes reveal that the (+)-(3S,4S) enantiomer HU-211 blocks N methyl-D-aspartate (NMDA) receptors in a stereospecific manner and that the interaction occurs at binding sites distinct from those of other noncompetitive NMDA antagonists or of glutamate and glycine. Moreover, HU-211 induces stereotype and locomotor hyperactivity in mice and tachycardia in rat, effects typically caused by NMDA receptor antagonists. HU-211 is also a potent blocker of NMDA induced tremor, seizures, and lethality in mice. This compound may therefore prove useful as a nonpsychoactive drug that protects against NMDA-receptor mediated neurotoxicity. PMID- 2556721 TI - Functional evaluation of cultured rabbit osteoblast-like cells. AB - For the improvement of the adult osteoblast culture, the osteoblasts of young adult rabbit endosteal from long bones were isolated by collagenase digesting separation. 0.1% of type-I collagen precoated culture flasks were used as substrate for isolated bone cell growth. Morphological examination of cultured cells under a phase-contrast microscope, SEM and TEM observations showed a structure similar to osteoblast in vivo. Histochemical examination of alkaline phosphatase demonstrated 97% purity of cultured osteoblasts. The presence of calcium deposit activity in cultured cells was demonstrated by Van Kossa stain. High activity of alkaline phosphatase and inorganic pyrophosphatase in cultured osteoblasts as determined by biochemical analysis. High calcium uptake in cultured osteoblasts was demonstrated by radioisotope labelled 45CaCl12. According to these methods, it was indicated that the cells isolated from young rabbit long bone endosteal were osteoblast-like and still maintained their biological function. Our system for culturing osteoblast-like cells is a successful attempt in growing bone tissue in vitro starting from isolated bone cells. Therefore, this modified method for bone cell culture on collagen precoated culture flasks could be used as the experimental model in studies concerning the osteoblasts in vitro. PMID- 2556720 TI - Purified, modified eel sodium channels are active in planar bilayers in the absence of activating neurotoxins. AB - A recent study showed that limited trypsin treatment of liposomes containing purified Electrophorus electricus sodium channels activates a sodium radiotracer flux. We now report that similarly treated sodium channels show voltage-gated, tetrodotoxin-sensitive and highly sodium-selective single-channel currents when incorporated into planar lipid membranes. The trypsinized channels opened repeatedly in bursts of several seconds duration, as would be expected for channels whose fast inactivation process had been removed. Furthermore, they have a higher conductance, different voltage-dependence of gating, and a remarkably higher selectivity (PNa/PK = 41) than sodium channels bound by batrachotoxin or other activating neurotoxins; these properties of the trypsinized channels are probably closer to those of channels in intact electrocytes. PMID- 2556722 TI - Purification and characterization of neuraminidase from Clostridium perfringens. AB - Neuraminidase (EC 3.2.1.18) has been purified from the culture medium of Clostridium perfringens ATCC 10543, through steps of gel filtration on Sephadex G 75 column, DEAE-cellulose DE 23 anion exchange chromatography, and isochromatofocusing. A homogeneous enzyme was obtained with a 7552-fold increase in specific activity to 295 units/mg protein. The yield was about 25%. The enzyme consists of a single polypeptide with a molecular weight of 69,000 as determined by SDS-polyacrylamide gel electrophoresis. Kinetic studies showed that Km is 1.5 mM for sialyllactose and Vmax is 0.41 mumole/min/ml at the enzyme concentration of 0.14 microgram/ml. The enzyme is stable at pH 5.2-8.0 with an optimal pH of 6.0. A concentrated solution of the purified enzyme was stable over one year at 4 degrees C. The purified enzyme hydrolyzed human alpha 1-acid glycoprotein completely; thus, it can be used in the clinical assay of N-acetylneuraminic acid in the serum. PMID- 2556723 TI - cDNA sequence of the long terminal repeat of a PrB strain of the Rous sarcoma virus. AB - The cDNA library of the polyA+ RNA from Prague B strain of RSV infected cells was constructed by the use of oligo(dT) primer and the reverse transcription method. The clones containing the LTR sequences were studied. The DNA sequences of several clones were studied and compared with the cDNA sequence of the long terminal repeat (LTR) region of a PrB strain of Rous sarcoma virus (RSV) which had previously been determined. By the analyses of several different cDNA clones, mutations were detected in the R region as well as in the U3 region of the LTR in two different cDNA clones. By comparing the DNA sequences of these cDNA clones with the original viral LTR, the error-prone hypothesis of the RSV reverse transcriptase was confirmed. PMID- 2556724 TI - Rotational behaviour produced by intranigral injections of bovine and human beta casomorphins in rats. AB - The biological activity of beta-casein derived beta-casomorphin peptides was evaluated by injecting bovine beta-casomorphin-5 (Tyr-Pro-Phe-Pro-Gly), the homologous sequence in human beta-casein (Tyr-Pro-Phe-Val-Glu) and the corresponding N-terminal tetrapeptides into the left substantia nigra of rats. Their ability to produce rotational behaviour was compared to that produced by three reference compounds, morphine, D-ala2D-leu5 enkephalin and U50,488H, ligands for mu, delta and kappa types of opioid receptors, respectively. The relative potencies of beta-casomorphins and morphine were compared to those tested in two in vitro assays for opioid activity: (1) inhibition of the electrically induced contraction of the isolated myenteric plexus-longitudinal muscle of the guinea-pig ileum and (2) displacement of 3H-dihydromorphine binding to brain membranes. The same ranking order of potency was found in all three assays, the peptides from human beta-casein being about 10-fold less potent than those from bovine beta-casein. The effects of both morphine and bovine beta casomorphin-5 in producing rotational behaviour were antagonized by naloxone; however, approximately 10-fold more naloxone was required to antagonize the beta casomorphin-5 effect than that of morphine. The present data are discussed in the light of the recent observation that high concentrations of beta-casomorphin-like peptides are found in the cerebrospinal fluid and plasma of women with postpartum psychosis. PMID- 2556725 TI - Fragments of ACTH affect electrophysiological signs of controlled stimulus processing in humans. AB - It has been proposed that the systemic administration of the 4-10 fragment of ACTH in humans affects primarily attention. In the present study, influences of ACTH 4-10 on event-related potential (ERP) indicators of attention were evaluated in healthy men. The influences were compared with those of an analog of the ACTH 4-9 sequence (HOE 427) which was expected to have an increased potency. Following an adaptation session, each of 20 healthy men was tested on four occasions in a double-blind study designed according to a latin-square. On each occasion, subjects received (iv) one of the following treatments: placebo, ACTH 4-10 (1 mg), HOE 427 (60 micrograms) and HOE 427 (200 micrograms). Treatments were administered 40 min prior to recordings of ERPs. ERPs were recorded while the subjects performed on a dichotic listening task paradigm. The various tone pips presented in this task elicit ERPs providing measures of controlled stimulus processing (Nd reflecting selectivity of attention, and P3) and automatic processing of stimulus deviance (mismatch negativity). ACTH 4-10 as well as 200 micrograms HOE 427 reduced Nd and also P3 amplitudes following attended stimuli. Smaller (non-significant) changes in the same direction were observed following 60 micrograms HOE 427. The results suggest an impairing influence of ACTH 4-10 and of HOE 427 on signs of controlled stimulus processing, particularly on the Nd. The analog appeared to be more potent than the endogenous 4-10 fragment. PMID- 2556726 TI - Mu-opioid component of the ethylketocyclazocine (EKC) discriminative stimulus in the rat. AB - The opioid receptor selectivity of the EKC discriminative stimulus was characterized in Fischer rats trained to discriminate 0.3 mg/kg EKC (SC) from saline in a two-choice discrete-trial avoidance paradigm. The putative kappa opioid receptor agonists EKC and U50,488H completely generalized with the EKC cue at doses of 0.3 and 10 mg/kg, respectively. The putative mu-opioid receptor agonists morphine (M) and fentanyl also dose-dependently generalized with the EKC stimulus. The generalization of M with EKC was not symmetrical, EKC and U50,488H produced little or no M-appropriate responding in rats trained to discriminate 3.0 mg/kg M (SC) from saline. This generalization pattern may reflect a lack of opioid receptor selectivity of the EKC stimulus. However, distinct mu-opioid and kappa-opioid components of the EKC cue could be identified using graded doses of naloxone in EKC-trained rats. The discriminative effects of morphine and fentanyl were blocked completely by doses of 0.1-1.0 mg/kg naloxone, whereas doses of naloxone 3-10 times greater were necessary to block the discriminative effects of EKC and U50,488H. These results suggest that EKC produces a complex discriminative stimulus with mu-opioid and kappa-opioid components that can be separated using antagonists such as naloxone. PMID- 2556728 TI - Intracellular signalling and regulation of gastric acid secretion. PMID- 2556729 TI - The nervous control of intraluminal pressure in the vomeronasal organ of the domestic ram. AB - In order to examine whether an aspiration mechanism exists in the vomeronasal organ (VNO) of the domestic ram, experiments were conducted in chloralose anaesthetized animals using acute electromanometry and electrophysiological techniques. Stimulation of the ipsilateral cervical sympathetic nerve activated a mechanism producing a negative (with respect to atmospheric pressure) intraluminal pressure which drew fluid into the VNO from the nasopalatine canal. The mechanism was under alpha-adrenergic control and did not seem to involve the carotid blood supply. An ipsilateral increase in intraluminal pressure occurred following electrical excitation of the maxillary trigeminal innervation. Thus a mechanism was demonstrated in the ram for conveying odours to the putative olfactory receptors in the VNO. PMID- 2556727 TI - Intravenous alprazolam challenge in normal subjects. Biochemical, cardiovascular, and behavioral effects. AB - Alprazolam, a novel benzodiazepine derivative is thought to be effective in the treatment of anxiety, panic, and depressive disorders. There is considerable interest in alprazolam's mechanism of action, particularly whether its profile of actions might resemble that of the alpha 2 adrenoreceptor agonist, clonidine. The present study assessed the biochemical, cardiovascular, and behavioral responses of healthy volunteers to acute intravenous infusions of alprazolam and placebo. Alprazolam reduced ACTH and cortisol while increasing growth hormone. There was a transient reduction in plasma norepinephrine and only modest effects on cardiovascular parameters. Subjects became quite sedated after intravenous alprazolam. This pharmacodynamic profile resembles that previously reported for traditional benzodiazepines, although alprazolam may be a more potent stimulator of growth hormone release. Alprazolam's effects on growth hormone resemble those of clonidine, but unlike clonidine, alprazolam has relatively little effect on plasma catecholamine and cardiovascular parameters. This suggests that alpha 2 mechanisms do not play a primary role in alprazolam's mode of action. Since alprazolam infusion affects three different measures (ACTH/cortisol, growth hormone, and plasma norepinephrine) thought to be dysregulated in depression, challenge with intravenous alprazolam may prove to be a useful "probe" in affective disorders. PMID- 2556730 TI - The modulation of force in isolated rat EGTA-treated anococcygeus muscle by phosphate, cyclic AMP and noradrenaline. AB - Saponin treatment is commonly used to permeabilize the surface membrane of muscle cells. Incubation of isolated rat anococcygeus for 4 h in a low Ca2+ and low Mg2+ solution resulted in the muscle responding to Ca2+, phosphate and cyclic AMP in a similar way to a saponin-treated muscle. However, unlike saponin-treated preparations, the maximal calcium-activated force was stable for many hours and the addition of noradrenaline caused contracture. These properties would permit this preparation to be used to study aspects of receptor-mediated excitation contraction coupling. PMID- 2556731 TI - Cholangiocarcinoma in two siblings with emphysema and alpha-1-antitrypsin deficiency. AB - Two siblings with homozygous ZZ alpha-1-antitrypsin deficiency were discovered to have primary liver tumours (both cholangiocarcinomas). This lends support to the view that alpha-1-antitrypsin deficiency plays a role in the development of some primary liver tumours. PMID- 2556732 TI - An ESR study of radical kinetics in L-alpha-amino-n-butyric acid hydrochloride containing L-cysteine hydrochloride. AB - On annealing at temperatures near 100 degrees C, carbon-centered radicals migrate to sulfur-centered radicals in X-irradiated crystals of L-alpha-amino-n-butyric acid hydrochloride, CH3CH2CH(NH3-Cl)COOH, containing L-cysteine hydrochloride, SHCH2CH(NH3Cl)COOH. Samples containing 0, 0.5, 1.0, and 1.5% L-cysteine hydrochloride were studied. When no cysteine is present, the carbon-centered radical formed by X irradiation, CH3CH2CHOOH, decays according to a second-order diffusion-controlled rate equation. In samples containing cysteine, the same carbon-centered radicals are formed, but on annealing, they migrate to cysteine, where a perithiyl radical, RSS, is formed. The transfer of carbon-centered radicals to perthiyl radicals follows a pseudo first-order rate equation with an activation energy of 1.15 eV. A decrease in the initial concentration of the carbon-centered radicals or an increase in the initial concentration of cysteine results in an increase in the transfer efficiency. The rate of growth of the perthiyl radical depends on both the initial concentration of cysteine and the initial concentration of carbon-centered radicals. The pseudo first-order rate constant increases when either the initial carbon-centered radical concentration increases or the initial cysteine concentration increases. The mechanism by which radicals move from one lattice site to another in the crystalline material is most likely hydrogen abstraction from a neighboring molecule. PMID- 2556733 TI - [Equilibrium radioisotope angiocardioscintigraphy in noninvasive diagnosis of chronic respiratory insufficiency]. AB - The role and utility of equilibrium radionuclide angiocardiography was estimated in non-invasive diagnosis of patients with Chronic Obstructive Pulmonary Disease (COPD). In 26 healthy subjects the average value of Right Ventricular Ejection Fraction (RVEF) was 54.8 +/- 7.2, versus 40.89 +/- 7.85 in 37 patients with COPD. Right heart catheterization was performed on 9 patients with COPD, randomly chosen, to analyze the correlation between RVEF values and average Pulmonary Artery Pressure (PAP) values: the correlation index was significantly high (r = 0.80). Moreover, an important agreement was observed between RVEF and PaCO2 values (r = 0.77). PMID- 2556734 TI - [Description of a radiation-induced extraskeletal osteogenic sarcoma. Therapeutic implications]. PMID- 2556735 TI - Sequential combination chemotherapy and radiotherapy in locally advanced non small cell carcinoma of the bronchus. AB - The feasibility, effectiveness and toxicity of a new four drug intensive combination chemotherapy regime has been assessed in 36 patients with inoperable non-small cell lung cancer. Three cycles of adriamycin, vindesine, ifosfamide and cisplatin were given to 36 patients followed by sequential radiotherapy to 25 patients. The overall response rate to chemotherapy was 33%, and median progression free interval was 6 months. Median survival was 8.5 months. Grade 4 WHO neutropenia was seen in 12 patients, there were three episodes of neutropenic fever and one death in which toxicity may have been contributory. Radiotherapy was not associated with serious toxicity. This study demonstrates that a significant response rate can be achieved with short intensive chemotherapy in unresectable non-small cell lung cancer and that it is feasible to combine this with radiotherapy. PMID- 2556736 TI - Pleural mesothelioma resulting from exposure to amosite asbestos in a building. PMID- 2556737 TI - [The value of CT and MRT in the diagnosis of cartilage-forming tumors]. AB - Accuracy in the assessment of lesion extension, specificity for histologic type and tumour status were reviewed in MRI and CT scans of 27 chondrosarcomas, 18 osteochondromas and 10 enchondromas and compared with the results obtained by plain radiography. No significant differences in the description of the lesion were found in chondrosarcomas and enchondromas between MRI and CT. In osteochondromas cartilage caps smaller than 5 mm and isodense to muscle were missed or not adequately delineated in CT. The specificity for the primary benign and malignant forms was equally high for CT and plain films, while MRI implies a much lower specificity due to the low sensitivity for matrix mineralisations. The criteria for malignant transformation of osteochondromas are visualised to better advantage by MRI. Solid ossified and small unmineralised recurrences of chondrosarcomas are detected by MRI with a higher sensitivity and specificity than by CT. PMID- 2556738 TI - [Computed tomography in the preoperative diagnosis of malignant laryngeal tumors]. AB - The pre-operative laryngoscopic and CT findings in the larynx in 50 patients with malignant laryngeal tumours were compared. Laryngoscopy showed an accuracy for T staging of 78.6% and CT an accuracy of 80%. In addition, CT complemented the endoscopic findings by demonstrating the depth of tumour invasion. The changes in individual anatomical portions of the larynx were compared, using both methods. CT and palpation showed equal specificity for lymph node staging, but CT is markedly superior in sensitivity. PMID- 2556739 TI - [Thoracic aorta dissection--the place of MRT and CT in the follow-up after prosthetic aortic replacement]. AB - Dynamic CT and MRT were performed in 21 patients who had undergone prosthetic replacement of the aorta because of dissection of the thoracic aorta. There is no difference between MRI and dynamic CT in the demonstration of a persistent intimal flap, the formation of a thrombus and the assessment of the aortic diameter in patients who underwent surgery for thoraco-abdominal aortic dissection. MRI, however, is superior in the identification of the true and false lumen and the demonstration of the distal anastomotic site. The major advantages of MRI are that there is no need for intravenous contrast agents, that it is highly sensitive to flow phenomena, and that it can demonstrate the aortic arch and the ascending aorta on sagittal oblique sections. The major disadvantages of MRI are the limited access to the patient during the procedure, and the inability to examine patients fitted with pacemakers or who are on assisted ventilation. Today, MRI is considered the method of first choice for the postoperative follow up of patients who underwent surgery for aortic dissection. PMID- 2556740 TI - [Diagnostic imaging following reconstructive surgery of the arteries of the legs. Angiodynography versus digital subtraction angiography]. AB - Fifty-three patients were examined by angio-dynography and angiography in a prospective study following vascular surgery of the lower extremities. The question of patency was correctly solved in all cases. A-V communications could be localised in eight out of nine cases. Evaluation of anastomoses is equally accurate by both methods; all angiographically demonstrated stenoses were also visible on sonography. Aneurysms at the anastomoses and peri-vascular fluid collections were better shown by angio-dynography, as might have been expected. The demonstration of the distal run-off, following femoro-crural bypass, presented a problem; in eight cases this could not be achieved because of technical difficulties. Surprisingly, angio-dynography proved superior in demonstrating large distal A-V shunts. Our experience so far indicates that angio dynography may be used as an alternative to angiography for certain specific indications (bypass patency, large A-V shunts and complications around the anastomosis). Demonstration of vessels, and particularly of distal femoral bypasses can be improved by using the "slow-flow technique". PMID- 2556741 TI - [The value of the transbrachial approach for arterial visualization using a 4-F catheter]. AB - To assess the safety of nonselective intraarterial digital subtraction angiography (DSA) performed from a transbrachial approach with 4-French catheters, the complications occurring in 158 examinations were studied. Two patients developed local thrombosis and in four patients there were temporary pulse deficits. Three brachial artery catheterisations were unsuccessful due to transient arterial spasm. Our data support the conclusion that nonselective transbrachial DSA is a safe and accurate method, easily employed in an outpatient population. PMID- 2556742 TI - Intra-arterial injection of papaverine in the decision of balloon dilatation of the iliac arteries. AB - In patients with low degree stenoses of the iliofemoral arteries, a discrepancy can occur between the clinical complaints and the severity of the angiographic lesions. Intra-arterial injection of 20 mg of papaverine by mimicking the need for augmented blood supply during exercise will help to objectify the clinical importance of a stenosis. Therefore, it will be of help in the decision of balloon dilatation of low-degree lesions. PMID- 2556744 TI - [Retroperitoneal xanthofibrogranulomatosis]. AB - Retroperitoneal xanthofibrogranulomatosis has so far been reported less than 50 times in the world literature. It has not been classified with retroperitoneal fibrosis or neoplasia. The clinical, radiological and histological features, and the problems of diagnosis, are discussed with reference to another case. The classification of the condition is considered in relation to the available literature. Diagnosis and possible treatment are also discussed. PMID- 2556743 TI - [The value of sonography in the pathologic evaluation of solid breast tumors]. AB - Six hundred and twenty patients were examined clinically and by sonography and mammography; 113 tumours were found and a histological diagnosis was attempted on the basis of a previously determined diagnostic protocol. There were 51 solid tumours; 48 were shown by mammography and 46 by sonography. Mammography proved more accurate in diagnosing malignant tumours (72% against 67%), whereas ultrasound was more precise in diagnosing benign tumours. Sonography found ten out of twelve histologically proven axillary lymph node metastases and was better than palpation (seven out of twelve). Mammography and sonography combined increased the accuracy of the pre-operative diagnosis of solid breast tumours. Ultrasound of the axilla should be used routinely in addition to clinical examination in order to detect non palpable lymph node metastases. PMID- 2556745 TI - [Hepatic and splenic abscesses in immunosuppressed patients]. AB - Morphologic characteristics of hepatosplenic abscesses using ultrasound and CT examinations in 13 immunosuppressed patients are presented. Additionally, the results of diagnostic ultrasound and CT guided biopsy procedures (n = 13) are reported. On sonograms, bacterial abscesses were exclusively hypoechoic lesions whereas patients with mycotic abscesses showed additionally target lesions and lesions presenting a "wheels-within-wheels" appearance. Thus, with some limitations, us might help to differentiate between fungal and bacterial abscesses. On CT, all patients presented uniformly with hypodense lesions. Follow up ultrasound studies showed these abscesses over periods as long as 24 months; biopsy proved some of these as fibrotic lesions without vital bacteria or fungi. PMID- 2556746 TI - Well-defined, dense and continuously spreading enhancement on single level dynamic CT of the liver: a characteristic sign of hepatic cavernous haemangioma. AB - The findings of single-level dynamic CT were reviewed in 83 lesions of cavernous haemangioma, 90 lesions of hepatocellular carcinoma, 29 lesions of metastatic tumour and 10 lesions of other hepatic masses in order to see the validity of characteristic findings of hepatic cavernous haemangioma. Well-defined, dense and continuously spreading enhancement (WDCSE) being defined as follows: 1. well defined contour of enhanced area, 2. the density of maximally enhanced area almost same as that of enhanced vessel and 3. continuously spreading enhancement continuing over 20 seconds, WDCSE was noted definitely in 47 lesions of cavernous haemangioma alone (57%; 77% over 3 cm, 64% between 2 and 3 cm, and 19% under 2 cm in diameter) and, suggestively in 16 lesions of cavernous haemangioma and 4 lesions of hepatocellular carcinoma, but in no lesions of other tumours. WDCSE did not show a high sensitivity for cavernous haemangioma but it had a 100% positive predictive value for the disease. If WDCSE is noticed even in oncology patients with hepatic mass, no further examination may be necessary. PMID- 2556747 TI - [Traction injuries of the brachial plexus: radiologic diagnosis using myelo-CT and MR]. AB - The exact radiographic localisation of supraganglionic lesions of the brachial plexus provides important information for the prognosis and clinical management of these injuries. The authors report on the results of enhanced CT scanning and MRI of the cervical spine in five patients with surgically proven root avulsions caused by traction injuries. All lesions were correctly diagnosed by enhanced CT scanning. MRI, by comparison, identified only about 70% of the neural lesions. PMID- 2556748 TI - [Injuries of the intracranial arteries in the angiogram]. AB - Damage to intracranial arteries was demonstrated angiographically in 24 patients with severe trauma to the skull and brain. The most common abnormalities were damage to the intima and traumatic carotid-cavernous fistulae (nine cases each). In addition, there were six traumatic aneurysms of the internal carotid artery, two extravasates from ruptured intracranial branches and one traumatic A-V fistula between the middle meningeal artery and the spheno-parietal sinus. The arterial damage was diagnosed either during the acute stage or after a latent period of days and weeks, if there were signs of intracranial bleeding or other evidence of vascular damage. Cerebral angiography in two or more projections is the method of choice for demonstrating even minor vascular lesions. PMID- 2556749 TI - [MRT in primary and tumor-induced syringomyelia]. AB - The MRI findings in 26 patients with primary syringomyelia and eight patients with a syrinx due to an intramedullary tumour were studied retrospectively. In general, the syrinx in the tumour cases resembled the appearance of primary syringomyelia. However, a specific and sensitive sign of tumour-induced syringomyelia was irregularity of the spinal cord close to the tumour (eight cases of eight), eccentric position of the lumen (seven of eight), and a diffuse increase of signal intensity on T2-weighted images (eight of eight). Primary syringomyelia also showed increased signal intensity on T2-weighted images outside the lumen, but this was closely related to the cavity. The use of Gd-DTPA improved demarcation of the tumour from the syrinx in all cases. Widening of the spinal canal and ectopia of cerebellar tonsils was a typical finding in primary syringomyelia. MRI proved a reliable method for distinguishing between a tumour induced syrinx and primary syringomyelia. PMID- 2556750 TI - Hypertrophic retromedullary venous drainage in spinal cord tumours: MR visualisation. AB - Pseudoangiomatous dilatation of retromedullary veins at myelography was noted in three cases of intramedullary spinal cord tumours. This concerned two haemangioblastomas at levels Th8 and L1, respectively, and an ependymoma of the conus terminalis. MRI accurately demonstrated the tumoural mass, together with the enlarged and tortuous draining perimedullary veins. In patients with dilated vascular shadows on myelography, MRI should be performed before medullary angiography to differentiate between spinal vascular malformations and intramedullary tumours with enlarged draining veins. PMID- 2556751 TI - [Laser ablation of the nucleus pulposus: optical properties of degenerated intervertebral disk tissue in the wavelength range 200 to 2200 nm]. AB - Percutaneous ablation of degenerative disk material by laser has been introduced recently as an alternative to enzymatic or mechanical discectomy. The optical properties of degenerative disk material were evaluated in the range from 200 to 2200 nm, which includes most of the laser wavelengths used for medical purposes. Remittance rate was determined in 16 post-mortem disks using a spectrophotometric unit and rate of absorption and scatter were calculated. High rates of diffuse remittance were found for classical laser wavelengths such as the argon or the Nd:YAG II laser indicating only low rates of absorption. In contrast, much higher rates of absorption were determined for excimer or mid-infrared lasers as well as for the Nd:YAG I (1320 nm), which appear more appropriate for laser discectomy than the Nd:YAG (1064 nm) laser. PMID- 2556752 TI - [Retained and displaced teeth detected by computed tomography]. AB - 55 patients with impacted displaced teeth were examined clinically and by orthopantomography and computerised tomography and the results compared. In the detection of impacted teeth, the results of orthopantomography and computerised tomography are comparable but the latter is considerably superior in demonstrating displacement, contact and absorption. PMID- 2556753 TI - [High-grade luminal obstruction of the descending aorta following aortitis]. PMID- 2556754 TI - [An aorto-bronchial fistula following prosthetic replacement of the descending aorta. A comparison of diagnostic possibilities]. PMID- 2556755 TI - Primary leiomyosarcoma of the lung originating in an emphysematous bulla. PMID- 2556756 TI - [Liver involvement by Fasciola hepatica: sonography and CT]. PMID- 2556757 TI - [Simultaneous occurrence of focal nodular hyperplasia and hemangioma of the liver -difficulties in differential diagnosis]. PMID- 2556758 TI - [Xanthogranulomatous pyelonephritis with urothelial carcinoma]. PMID- 2556759 TI - Radiological findings in an unusual focus of extramedullary haematopoiesis in Polycythemia vera. PMID- 2556760 TI - [Computed tomographic findings in hemangiolipomatosis of the bones]. PMID- 2556761 TI - [CT diagnosis of a diffuse episcleritis]. PMID- 2556762 TI - [Polymyositis in the Sharp syndrome. Magnetic resonance tomographic, light and electron microscopic findings]. PMID- 2556763 TI - [Ultrasound-guided percutaneous injection of alcohol. Is it an elective therapy in small diameter nodular hepatocarcinoma associated with cirrhosis?]. AB - The authors discuss percutaneous alcohol injection as elective therapy for small hepatocellular carcinoma (less than 5 cm) in liver cirrhosis. Thirteen patients were classified as Child C cirrhosis. Thirty-six months actuarial survival is 60%. These results are similar in patients selected for surgical resection. PMID- 2556764 TI - [Chemotherapy of cancer of the lung]. AB - Based on the present experience, it is suggested that CDDP plus VP16 is associated with a reproducible 30-40% objective response rate in NSCLC (Non-Small Cell Lung Cancer). Although symptomatic relief is not rare in responding patients, serious morbidity from chemotherapy may also occur. Overall, the duration of the response is not very long (the median duration is approximately 6 8 months) and only modestly influences the survival of responding patients (the median survival in non responders is usually 4-5 months). In addition, due to such factors as the relatively low response and the serious morbidity from treatment, the overall survival and quality of life for these patients has not substantially improved. As for as SCLC (Small Cell Lung Cancer) is concerned, the impact of chemotherapy despite the introduction of VP16, carboplatin and ifosfamide, has remained essentially unchanged for more than ten years. Maintenance chemotherapy, alternating non cross resistant combinations and intensive treatments, with presently available drugs, have failed to improve the prognosis of patients with SCLC. Combined radio chemotherapy also fails in this regard. Truly limited disease should definitely be treated with chemotherapy followed by surgery, as the potential for prolonged survival looks realistic. Therapy of more extensive disease still remains a challenge; despite the failure of early and late intensification programs, more aggressive initial multi-drug therapy represents an alternative approach. PMID- 2556765 TI - Sites of persistence of feline calicivirus. AB - Various tissues were collected from eight cats persistently infected with feline calicivirus (FCV) strain 255 to determine the sites of viral persistence. Tissues were tested by virus isolation and an immunohistochemical technique in which infected cells were detected in formalin-fixed, paraffin-embedded tissue sections using rabbit antiserum to FCV 255, a biotinylated second antibody and streptavidin-peroxidase. Virus was detected by one or both techniques in tonsillar tissues of each animal, and not in other samples. Infected cells were detected in samples from six of eight kittens, and in each animal were few in number, and were cells of the superficial tonsillar epithelium or the stratum germinativum of the adjacent fossa mucosa. Transmission electron microscopic examination of tissues from three of the cats revealed calicivirus-like particles in cells similar to those identified immunohistochemically. These results confirm that the tonsillar region is the major site of FCV persistence and indicate that virus replication during persistence is confined to the surface epithelium of the tonsil and adjacent fossa mucosa. PMID- 2556767 TI - [Application of a gas chromatography-mass spectrometer (GC-MS) to the multiple inert gas elimination technique: multiple inert gas measurement with a GC-MS at trace level]. PMID- 2556766 TI - Comparative effects of proligestone and megestrol acetate on basal plasma glucose concentrations and cortisol responses to exogenous adrenocorticotrophic hormone in cats. AB - Cats were given megestrol acetate (MA, 5 mg once daily for 14 days), subcutaneous proligestone (PRG, 100 mg on two occasions one week apart) or subcutaneous saline (1 ml as for PRG). In cats given saline (n = 6) basal cortical concentrations, cortisol concentrations after adrenocorticotrophic hormone (ACTH) administration and fasting blood glucose concentrations did not change significantly during the following seven weeks. Cats given MA (n = 7) developed suppression of basal and ACTH-stimulated cortisol concentrations and fasting hyperglycaemia during treatment. Effects on cortisol persisted for two weeks after MA dosage ceased. In cats given PRG (n = 7), basal cortisol concentrations were reduced overall, but only three cats had persistently suppressed post-ACTH cortisol concentrations. Adrenal suppression continued for 14 weeks in one of these and for at least 22 weeks in two cats. Fasting blood glucose concentrations were unchanged in PRG treated cats. PMID- 2556768 TI - [New quantitative method for non-invasive monitoring of tissue blood oxygenation by near infrared spectrophotometry]. AB - The near infrared absorption spectra was measured with the transmitted light through rat brain under the various condition. The absorbance changes below 780 nm were attributable to hemoglobin (Hb) in the brain tissue, whereas those above 780 nm were associated with both Hb and cytochrome oxidase. To eliminate possible interference from cyt. oxidase, two wavelengths, 750 nm and 780 nm, were used to measure Hb oxygenation in the tissue. The absorbance changes in human blood cell suspensions were measured with changes in hematocrit values in the optical cuvette. At two wavelengths 750 nm and 780 nm, there was a linear relationship between absorbance changes and hematocrit values. Through these in vitro studies, the following equation (1) and (2) were obtained to monitor quantitatively the changes of oxy-Hb content (delta Hb O2) and total-Hb content (delta Hb Vol.) in the living tissue. These are (1) delta Hb O2 = -1.15 delta A 750 + 1.39 delta A 780, (2) delta Hb Vol. = -0.29 delta A 750 + 0.59 delta A 780. The studies using these equations showed that the oxy-Hb content in the brain was decreased as the O2 concentration in inspired gas was lowered with a half of Hb deoxygenated at 7% O2. The reliability of these equations was examined under the various conditions in situ such as CO2 inhalation, intravenous injection of Ca2+-blocker nicardipine, hemorrhage and retransfusion. These results confirmed that these equations derived from in vitro studies, were successfully applied to the in situ measurements of the oxygenation state of Hb in the living tissues. PMID- 2556769 TI - [The liver: from divination to molecular biology]. PMID- 2556770 TI - Human monoclonal antibodies: prospects for the therapy of cancer. AB - Human monoclonal antibodies have been produced in a number of laboratories to a number of cell surface and cytoplasmic antigens. Although currently more difficult to produce than murine monoclonal antibodies, human monoclonal antibodies provide several potential advantages over conventional murine monoclonal antibodies in the therapy of human cancer. We review the human monoclonal antibodies produced to human tumors and describe our clinical experience with the use of human monoclonal antibodies in the treatment of recurrent cutaneous melanoma. PMID- 2556771 TI - [Endorphins 1989: neuroendocrine aspects]. AB - Endorphins represent a family of brain peptides having an opioid core structure responsible for their morphinomimetic activity. We present an overview of the biology of endorphins including their origin, neuroanatomical distribution, regulatory mechanisms and interaction with opioid receptors taking neuronal cellular function in situ as our focal point of interest. Differential biochemical processing and regulatory mechanisms both at the pre- and post translational level in the various brain areas contribute to the biochemical and functional diversity of peptides generated from common precursors in keeping with the selective needs and functions of neuronal circuits. Physiological significance requires interaction of the various endogenous ligands thus generated with appropriate receptors, sometimes located well beyond the distance of a synaptic cleft as demonstrated by the diverging ligand/receptors neurochemical topographical maps. Physiological analysis reveals a multitude of potential interactions between peptides and classical neurotransmitters in part as a result of their common subcellular location within individual synaptic vesicles but also in relation to qualitatively different post-synaptic effects and metabolism, enhancing the complexity of their potential role in neuronal function. Endorphins may contribute to major neuroendocrine systems such as analgesia, endocrine and autonomic function, mental function and behavior. Their implication in various neuropsychiatric and endocrine clinical disorders will be briefly reviewed. PMID- 2556772 TI - [Clinical forms of inclusion body myositis: 12 cases]. AB - Inclusion body myositis is now a well-known disease but its incidence is underestimated. We report 12 cases with clinical heterogeneity. Three groups of patients could be described. The first one corresponded to asymmetrical muscle involvement and distribution with a slow clinical course (4 cases). The second was characterized by a polymyositis-like syndrome (3 cas), but steroid therapy was ineffective. The last group mimicked a chronic spinal muscular atrophy (4 cases). One patient showed a scapuloperoneal syndrome. Both myopathic and neurogenic EMG patterns were present in 6 patients; a neurogenic pattern was found in 4 cases and a myopathic pattern in 2 cases. In all patients, muscle biopsies showed rimmed vacuoles with eosinophilic inclusions. In 9 cases ultrastructural studies displayed abnormal filaments of 15-18 nm in diameter in the vacuoles. Intranuclear filaments were rarely observed. The significance of the filaments is unknown and their specificity is doubtful because they are present in other myopathies with rimmed vacuoles (some distal myopathies and oculopharyngeal muscular dystrophies). Finally a rich inflammatory exudate was present in 8 patients only. PMID- 2556773 TI - [Hypothesis: multinucleated giant cells in AIDS neuropathology]. AB - Data of the literature about frequency and specificity of giant cells during AIDS encephalitis are conflicting. These discrepancies could be explained by several factors. Envelope glycoprotein Gp 120, when free, induces the formation of giant cells through the fusion of cerebral monocytes. The concentration of this protein depends on the level of its production, function of the quantity and of the maturation of the HIV, and on the rapidity of its catabolism, due to anti-Gp 120 antibodies. On the other hand, the tropism of the virus for monocytes and lymphocytes could vary; this would lead to the formation of giant cells with a different life span. The fusion capability of HIV-2 might be lower on account of the smaller size of its Gp 120. The frequency of mutations in HIV lentivirus leading to the synchronous development of several different clones of HIV-1 in the same patient and the associated infectious diseases, due to risk factors common to the diseased population, current endemic or opportunistic agents, could explain the variability of the density of giant cells according to the country where the patients live. PMID- 2556774 TI - [Acute and reversible axonal polyneuropathy in post-leptospirosis]. AB - A 53 year old woman presented, one month after an anicteric leptospirosis, an acute, asymmetrical, sensorimotor polyneuropathy involving the lower limbs. Electrophysiological study showed evidence of severe denervation, with normal motor nerve conduction velocities, indicating an axonal degeneration. Neuro muscular biopsy showed signs of wallerian degeneration and perivascular infiltrates of epineural vessels. She received a corticosteroid therapy during 6 months and there was a nearly complete clinical recovery. PMID- 2556775 TI - [Clinico-pathologic case of slowly progressive herpes simplex encephalitis without temporal necrosis]. AB - A case of herpes simplex encephalitis (HSE) is reported. The patient experienced short term memory disorders and irritability progressing over 3 months, without seizures or fever. The CSF was normal. CT showed a small low density area in the right posterior orbito-frontal region. At post-mortem examination, one month later, the temporal cortex appeared largely spared by necrosis, which involved the posterior orbito-frontal areas. Cowdry type A inclusions, herpes virus like particles and fluorescent reaction with HSV1 monoclonal antibodies strongly supported the diagnostic. Such atypical cases of long duration have apparently seldom been reported. They suggest that HSE should be considered in the differential diagnosis of a subacute encephalopathy. PMID- 2556776 TI - Diagnosis and management of diseases affecting the motor unit in infancy. PMID- 2556777 TI - Current status of the clinical use of heparin fractions. AB - In recent years, numerous low molecular weight heparin (LMWH) fractions have become available for clinical use, and the widespread registration of some of these products is expected soon also in Italy. An increased benefit/risk ratio concerning efficacy and safety has been shown in animal models as compared to standard heparin. In order to assess whether there is sufficient evidence supporting their use in replacement of unfractionated heparin, a review of the results of the main clinical studies is performed, discussing LMWH effectiveness and bleeding risk separately in the different clinical applications. PMID- 2556778 TI - [The sympathetic skin response: neurophysiological basis, normal values, clinical applications]. AB - The sympathetic skin response (SSR) represents the momentary change in skin potential reflexively evoked by a variety of arousal stimuli. Although sudomotor unmyelinated fibres are the final efferent pathway of SSR, little is known about the afferent and central components. SSR was recorded using different classes of stimuli in 20 controls, 15 peripheral neuropathy and 13 stroke patients. In controls SSR latencies changed significantly with different recording sites but not with different stimulation sites. Additionally, ischemic conduction block of the arm abolished SSR recorded at the hand after median nerve stimulation. In 1 patient with subacute ganglionitis and in 3 with demyelinating neuropathies the SSR could be elicited with deep inspiration but not by electric stimulation. These results suggest that myelinated fibres serve as afferents for SSR. In stroke patients SSR was absent bilaterally after stimulation of the paralyzed side but present after stimulation of the normal one. Therefore in humans the cortex seems to have a suprasegmental excitatory influence on SSR. The above findings imply that an unobtainable SSR by electric stimulation may be due not only to dysfunction of the autonomic efferent nerve fibers, but also to involvement of sensory afferents or suprasegmental structures. PMID- 2556779 TI - [Mechanism of platelet aggregation and mode of action of platelet antiaggregants]. AB - Platelets play an important role in arterial thrombosis. Following the adhesion of platelets to an injured vascular wall, platelet activation occurs, involving calcium fluxes, phosphoinositol metabolism, protein phosphorylation, arachidonate cascade. This leads to the formation of the fibrinogen membrane receptor, the IIb/IIIa glycoprotein complex. Platelet aggregation results from the binding of fibrinogen and other adhesive proteins to the IIb/IIIa complex between several platelets. Numerous drugs can interfere with platelet function. Among these, only aspirin and ticlopidine have been shown to be effective in controlled trials. Aspirin, by inhibiting cyclo-oxygenase, blocks thromboxane A2 formation. Ticlopidine, by inhibiting fibrinogen binding to the complex, is a potent antiaggregation agent. PMID- 2556780 TI - [Anatomo-pathologic ovarian cancers]. AB - Ovarian cancers constitute a group of tumours characterized by a variety of histological patterns. The classification used is that of the W.H.O. Common epithelial account for two-thirds of all ovarian cancers. In this group, tumours of borderline malignancy should be distinguished from adenocarcinomas. Since these lesions are associated with an unusually good prognosis, it is of major importance to rule out an infiltration of the ovarian stroma. Extra-ovarian spread of borderline tumours probably results from a multifocal process which is not associated with the poor prognosis of metastatic involvement. Tumours of the mesenchyma and sex cords are less frequent, as are germ cell tumours. For the latter, preoperative assays of serum markers can be of help in the diagnosis and management of the cases. PMID- 2556781 TI - Discrepancy between serum concentrations of 1,25-dihydroxyvitamin D metabolites measured by radio-immunoassay and thymus radioreceptor assay during vitamin D2 treatment. AB - A radio-immunoassay (RIA) for determination of 1,25-dihydroxycholecalciferol (1,25(OH)2D3) and 1,25-dihydroxyergocalciferol (1,25(OH)2D2) in serum was compared with a competitive protein binding assay using calf thymus receptor to clarify the comparability of measurements in subjects treated with either vitamin D2 or vitamin D3 (4000 IU per day for 8 weeks). Before and during treatment with vitamin D3 the two assay techniques were concordant, but during vitamin D2 treatment the serum concentrations of 1,25(OH)2D2 and 1,25(OH)2D3 were higher when measured by RIA. The study demonstrates that during vitamin D2 treatment unknown compounds, recognized only by the antiserum, co-elute with 1,25(OH)2D2 and 1,25(OH)2D3 in high pressure liquid chromatography. It can be concluded that the radio-immunoassay used here cannot replace the calf thymus receptor assay. PMID- 2556782 TI - Na,K-ATPase activity in rectal mucosa of children with ulcerative colitis and Crohn's disease. AB - Mucosal Na,K-ATPase activity was studied in rectal biopsy specimens from 19 children with ulcerative colitis (UC) (mean age, 13 years) and 4 children with Crohn's colitis (mean age, 14 years) and compared with biopsy specimens from 12 control children (mean age, 12 years). The Na,K-ATPase activity was significantly decreased in UC with severe rectal inflammation compared with UC in remission or with children with unspecific symptoms and normal mucosa (p less than 0.001, respectively). A higher enzyme activity was shown with age in the group with normal rectal mucosa and no evidence of inflammatory bowel disease (n = 17). The decreased Na,K-ATPase in UC with severe rectal inflammation might contribute to the diarrhoea by impairment of sodium transport. PMID- 2556783 TI - Effect of diethyldithiocarbamate on gastric and duodenal mucosal blood flow and vascular permeability in rats. AB - Diethyldithiocarbamate (DDC), an inhibitor of superoxide dismutase, produced ulceration in the proximal duodenum of rats, and erosions or ulcers in the stomach. DDC given s. c. at 1.5 g/kg increased duodenal and gastric vascular permeability, and reduced blood flow. The decrease in gastric blood flow caused by DDC was attenuated by pretreatment with superoxide dismutase, and significantly inhibited by a xanthine oxidase inhibitor, allopurinol. It is suggested that the superoxide and other radicals damage the mucosal vasculature, and play a role in the pathogenesis of gastric and duodenal mucosal lesions. PMID- 2556784 TI - Possible mechanisms of diethyldithiocarbamate-induced gastro-duodenal mucosal damage in rats. AB - A single s.c. injection of diethyldithiocarbamate (DDC, 1 g/kg) induced not only gastric but also duodenal mucosal damage in rats. DDC induced marked decreases in gastric acid output, gastro-duodenal mucosal blood flow and transmucosal potential difference prior to the development of mucosal lesions. Superoxide dismutase activity in the gastro-duodenal mucosa was also inhibited, while catalase and glutathione peroxidase activities gradually increased after the administration of DDC. These results suggest that a decrease in mucosal defensive mechanisms plays an important role in the development of DDC-induced gastro duodenal mucosal damage and that oxygen-derived free radicals may also participate in the development of this mucosal damage. PMID- 2556785 TI - A new model of duodenal ulcers induced in rats by diethyldithiocarbamate, a superoxide dismutase inhibitor. AB - Repeated administration of diethyldithiocarbamate (DDC: 750 mg/kg, s.c.), a superoxide dismutase (SOD) inhibitor, to fed rats induced ulcers in the duodenum with less lesion in the stomach. DDC not only reduced basal acid output but also impaired duodenal alkaline secretion under both basal and acid-stimulated conditions. The duodenal ulcers induced by DDC were significantly prevented by either allopurinol, SOD or dmPGE2 at the doses which significantly reversed the inhibited alkaline responses caused by DDC. The pathogenesis of DDC-induced duodenal ulcers may involve impairment of duodenal alkaline secretion, probably caused by insufficiency of antioxidant machinery in the mucosa. PMID- 2556786 TI - Prolylhydroxylase and collagenase activities in the healing of acetic acid induced gastric ulcers in rats and the effects of cimetidine and elcatonin. AB - We examined the changes in prolylhydroxylase (PH) and interstitial collagenase (CA) activities in the healing of acetic acid-induced gastric ulcers in rats. Gastric ulcers were induced by subserosal injection of acetic acid, and confirmed by endoscopy on the third day. At the acute stage of ulceration, PH activity was not high but CA activity was increased. At the healing stage, PH activity was increased but CA activity gradually decreased to a normal level. Cimetidine did not effect PH activity but tended to increase CA activity in the healing. On the contrary, elcatonin increased PH activity but had little effect on CA activity. The above enzymes seems to be indispensable in the healing of gastric ulcers and effects of antiulcer agents on these enzymes must be considered. PMID- 2556787 TI - Gastric lesions induced by kainic acid injection into the dorsal motor nucleus of the vagus nerve in rats. AB - Kainic acid injection into the dorsal motor nucleus of the vagus nerve (DMN) induced lesions in the glandular stomach of rats. A decrease in mucus production seemed to play an important role in the pathogenesis of the kainic acid-induced gastric lesions. On the other hand, by means of horseradish peroxidase tracing method, the original cells projecting to the DMN were identified in the central nucleus of the amygdala, several hypothalamic nuclei and the insular cortex. Considering that kainic acid is a long-acting neuronal excitant, it was suggested that the continual stimulation of DMN neurons facilitated gastric ulcer formation. PMID- 2556788 TI - Inhibition of H+,K+-ATPase by methyl(E)-2-(3,4-dimethoxystyryl)-benzimidazole-4 carboxylate (ALE-36). AB - ALE-36, as well as omeprazole and SCH 28080, markedly inhibited the [14C]aminopyrine (AP) accumulation induced by dibutyryl cyclic AMP (dbcAMP) and H+,K+-ATPase activity in a concentration-dependent manner. The inhibitory effect of omeprazole on the dbcAMP-induced [14C]AP accumulation was reversed by treatment with beta-mercaptoethanol, but those of ALE-36 and SCH 28080 were not. ALE-36 and SCH 28080 did not inhibit dog renal Na+,K+-ATPase activity, while omeprazole and ouabain did inhibit this enzyme activity. These results suggest that the inhibitory action of ALE-36 on acid secretion is due to the specific inhibition of gastric H+,K+-ATPase, the manner being different from in the case of omeprazole. PMID- 2556789 TI - Roles of endogenous leukotrienes and prostaglandins in the healing of gastric ulcers induced by acetic acid in rats. AB - We investigated the roles of endogenous leukotrienes (LTs) and prostaglandins (PGs) in the healing of gastric ulcers induced by acetic acid in rats. The mucosal levels of LTB4 and sulfidopeptide LT at the ulcer edge had increased by one day after the induction of ulcers. AA-861, a selective inhibitor of 5 lipoxygenase, did not affect the ulcer healing. Indomethacin delayed the healing. Cimetidine did not affect this delay, but ornoprostil, a PGE1, derivative, prevented it. These results suggest that endogenous LTs are not related to the healing of gastric ulcers and that a deficiency in endogenous PGs may be involved in the persistence of gastric ulcers. PMID- 2556790 TI - Pathogenesis of platelet-activating factor-induced gastric mucosal damage in rats. AB - Platelet-activating factor (PAF), given intravenously, induced erosions and hyperemia to the rats stomachs. Gastric mucosal blood flow was decreased and thiobarbituric acid (TBA) reactants (an index of lipid peroxidation) in the gastric mucosa were increased 10 min after PAF injection. Superoxide dismutase plus catalase reduced the gastric mucosal lesions and TBA reactants, but had no influence on gastric mucosal blood flow. A reduction in the number of circulating polymorphonuclear leukocytes (PMN) reduced the gastric mucosal damage and TBA reactants, and inhibited the decrease in gastric mucosal blood flow, as observed 30 and 60 min after PAF injection. PAF induced superoxide production by rat PMN and enhanced that stimulated by opsonized zymosan or phorbol myristate acetate. These results suggest that microcirculatory disturbance and oxygen-derived free radicals generated by PMN play important roles in gastric mucosal lesions induced by PAF. PMID- 2556791 TI - Role of lipid peroxidation and oxygen radicals in compound 48/80-induced gastric mucosal injury in rats. AB - Repeated administration of a small dose of compound 48/80 induced acute gastric mucosal injury in rats. Thiobarbituric acid (TBA) reactants in the gastric mucosa were significantly increased and serum alpha-tocopherol was significantly decreased after the treatment. The total area of gastric lesions and the increase in TBA reactants in the gastric mucosa were significantly reduced by pretreatment with superoxide dismutase (SOD) and/or catalase, allopurinol and anti-rat polymorphonuclear leukocytes (PMN) antibody. Lipid peroxidation and oxygen radicals derived from both the xanthine-xanthine oxidase system and PMN may be involved in the pathogenesis of compound 48/80-induced gastric mucosal injury. PMID- 2556792 TI - The efficacy of tenoxicam in patients suffering from rheumatoid arthritis (including assessments of quantified articular scintigraphic data). AB - Every doctor faced with the task of testing a drug is well aware of the difficulty of making his or her results objective. In designing test procedures, methods need to be found which are unaffected by the subjective views of either the testing doctor or the patients undergoing the treatment. This study was designed taking this into account, using scintigraphic joint measurements in patients with chronic rheumatoid arthritis (RA) to record comparable data. Even this method, however, despite its indisputable scientific accuracy, requires clinical interpretation. PMID- 2556793 TI - One Wilms' tumor gene is cloned; are there more? PMID- 2556794 TI - Is the AIDS epidemic slowing? PMID- 2556795 TI - Inhibition of antigen-induced lymphocyte proliferation by Tat protein from HIV-1. AB - The purified human immunodeficiency virus type-l (HIV-l) Tat protein inhibited lymphocyte proliferation induced by tetanus toxoid or Candida antigens by 66 to 97% at nanomolar concentrations of Tat. In contrast, Tat did not cause a significant reduction of lymphocyte proliferation in response to mitogens such as phytohemagglutinin or pokeweed mitogen. Inhibition was blocked by oxidation of the cysteine-rich region of Tat or by incubation with an antibody to Tat before the assay. A synthetic Tat peptide (residues 1 to 58) also inhibited antigen stimulated proliferation. Experiments with H9 and U937 cell lines showed that Tat can easily enter both lymphocytes and monocytes. The specific inhibition of antigen-induced lymphocyte proliferation by Tat mimics the effect seen with lymphocytes from HIV-infected individuals and suggests that Tat might directly contribute to the immunosuppression associated with HIV infection. PMID- 2556796 TI - Molecular cloning of the thyrotropin receptor. AB - The pituitary hormone thyrotropin, or thyroid-stimulating hormone (TSH), is the main physiological agent that regulates the thyroid gland. The thyrotropin receptor (TSHR) was cloned by selective amplification with the polymerase chain reaction of DNA segments presenting sequence similarity with genes for G protein coupled receptors. Out of 11 new putative receptor clones obtained from genomic DNA, one had sequence characteristics different from all the others. Although this clone did not hybridize to thyroid transcripts, screening of a dog thyroid complementary DNA (cDNA) library at moderate stringency identified a cDNA encoding a 4.9-kilobase thyroid-specific transcript. The polypeptide encoded by this thyroid-specific transcript consisted of a 398-amino acid residue amino terminal segment, constituting a putative extracellular domain, connected to a 346-residue carboxyl-terminal domain that contained seven putative transmembrane segments. Expression of the cDNA conferred TSH responsiveness to Xenopus oocytes and Y1 cells and a TSH binding phenotype to COS cells. The TSHR and the receptor for luteinizing hormone-choriogonadotropin constitute a subfamily of G protein coupled receptors with distinct sequence characteristics. PMID- 2556797 TI - Steroid binding at sigma-"opioid" receptors. PMID- 2556798 TI - In vitro activity of some newer quinolone compounds. AB - The quinolones are a group of antimicrobial agents that act by inhibiting bacterial DNA gyrases, enzymes essential in DNA replication. Several newer quinolone agents have been introduced recently. These are broad spectrum agents which may be administered orally. In-vitro susceptibility testing of five quinolone agents namely norfloxacin, pefloxacin, enoxacin, ofloxacin and ciprofloxacin against recent clinical bacterial isolates at the General Hospital Kuala Lumpur was performed. The results confirm the broad spectrum and high activity of these agents against these isolates which included Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus. The quinolones would provide valuable alternatives in the treatment of infections caused by organisms resistant to the more commonly used antibiotics. PMID- 2556799 TI - Hypocalcemia and an inappropriate endocrine response in osteoblastic metastatic breast cancer. AB - Hypercalcemia in association with skeletal metastases is common; hypocalcemia in this clinical setting is unexpected, though it has also been described, most commonly with primary lesions of the breast or prostate. In a subset of hypocalcemic patients with breast cancer, there is an inappropriate endocrinologic response as evidenced by a relative hypoparathyroidism and an elevation in the serum level of calcitonin. We have described a representative case and reviewed the literature. PMID- 2556800 TI - Characterization of human IFN-gamma response using somatic cell hybrids of hematopoietic and nonhematopoietic origin. AB - A panel of 27 rodent-human somatic cell hybrids composed of cells of hematopoietic (nonadherent cells) and nonhematopoietic origin (adherent cells) was used to identify the chromosomes involved in the biological response to human IFN-gamma (Hu-IFN-gamma). We found that the stimulation of class-I histocompatibility antigen expression correlates with the presence of human chromosomes 6 and 21 in adherent cell hybrids, while human chromosome 6 alone is sufficient in nonadherent hybrids. Scatchard analysis of the binding of radiolabeled Hu-IFN-gamma to nonadherent cell hybrids gave a Kd value similar to that found on human cell lines. Induction of a reporter gene placed under the transcriptional control of the interferon responsive sequence (IRS) in adherent cell hybrids requires both chromosomes 6 and 21. The antiviral protection by Hu IFN-gamma in adherent cell hybrids was reached at physiological doses (2 units/ml) when human chromosomes 6 and 21 were present, while higher doses of Hu IFN-gamma (5000 units/ml) were required for hybrids lacking chromosome 21. Thus, we demonstrate that differences exit in the response to Hu-IFN-gamma depending on the origin of the cell type. PMID- 2556801 TI - Long-range activation of transcription by SV40 enhancer is affected by "inhibitory" or "permissive" DNA sequences between enhancer and promoter. AB - The transcriptional enhancer effect is used in many, if not all, organisms for remote control of gene transcription. An enhancer DNA can dramatically stimulate transcription of a linked gene from positions either 5' or 3' to the gene. Both the proximal promoter and the distal enhancer sequences are binding sites for transcription factors. Interaction between promoter and enhancer is mediated by these factors, presumably via looping out of the intervening DNA. Here we report that the extent of remote activation by an enhancer depends on characteristics of that intervening DNA. Using Beta-globin and SV40 T-antigen test genes, we show that the effect of an SV40 enhancer is transmitted to the responsive promoter, with little or no loss of efficiency, through certain segments of mammalian DNA derived from rabbit beta-globin or mouse alpha-globin gene regions. By contrast, a strong reduction of enhancer activity is observed with certain spacer segments of prokaryotic DNA (from plasmid pBR322 or phage lambda) or sequences of high (G + C) content from eukaryotic genes. We have analyzed more closely sequences that are more or less permissive for transmission of the transcriptional enhancer effect. It appears that these permissive sequences generally have a high (A + T) content and notably a very low abundance of CpG dinucleotides. By contrast, (G + C)-rich DNA segments with high local densities of CpG were the most deleterious for long-range enhancer action. We note that the latter sequence composition is typical for "CpG islands" of many mammalian genes, including housekeeping genes and the human alpha-globin gene. This may be related to the finding that promoters of most cell type-specific genes, whose activity depends on a strong enhancer, do not contain CpG islands. Most likely, the spacer DNAs of typical cell type-specific genes have evolved to allow maximal transmission of the enhancer effect. PMID- 2556802 TI - Long-range restriction map around 11p13 aniridia locus. AB - Using two random DNA markers, and pulsed field gel electrophoresis, a 1.5-Mb physical map surrounding the 11p13 aniridia locus (AN2) has been assembled. The map was constructed using a combination of single- and double-restriction digests on DNA from normal controls and a patient transmitting familial aniridia. The aniridia patient has a chromosome translocation and the two DNA markers flank the breakpoint. This 11p13 breakpoint lies no further than 100 kb from the DNA marker 1104 (D11S95), located on the centromeric side of the breakpoint. Two CpG islands, separated by 550 kb and flanking the translocation, suggest an upper limit to the size of the gene. PMID- 2556803 TI - [Cytomegaloviruses and synthetic acyclic nucleosides: specificity of ganciclovir]. PMID- 2556804 TI - [Detection of antibodies to cytomegaloviruses, herpes simplex viruses and varicella-zoster viruses in persons positive for human immunodeficiency virus antibodies]. AB - Forty-seven anti-HIV positive sera and 47 sera of normal individuals were analyzed to the presence of IgG and IgM antibodies versus CMV, HSV and VZ. IgG antibodies in comparison to CMV were found in 27 cases (57.44%); in comparison to HSV in 40 (85.1%) subject and to VZ in 33 anti-HIV positive subjects. The mean absorbance value was higher in anti-HIV positive cases than in controls. IgM antibodies in comparison to CMV were found in 3, and in comparison to HSV in 2 anti-HIV positive sera. No IgM antibodies were found in the control group. PMID- 2556805 TI - Asymptomatic rotavirus infections among normal Indian children in Chatsworth, Durban. AB - During a 27-month survey in Chatsworth, Durban, serum from 1,041 normal Indian children, ranging in age from birth up to 13 years, was tested for the presence of anti-rotavirus antibodies by means of a complement fixation test. It was found that from an initial high positivity rate of 47.7% in the newborn, there was a sharp drop to 23.8% in the 1-2-month age group (P = 0.0009). This low positivity rate was maintained up to the age of 9-11 months, after which it rose to 46.4% in the 12-17-month age group (P = 0.0006). There was a further significant rise between the 2-3-year and 4-5-year age groups, probably reflecting rotavirus infections in nursery school and/or in the home, the latter being nosocomially acquired from younger siblings. Stool samples were obtained from 829 of the above subjects: overall, 16.2% were positive for rotavirus antigen by enzyme-linked immunosorbent assay; the highest rate (29.5%) of asymptomatic rotavirus infection was in the 12-14-month age group. The data indicate that asymptomatic infection with rotavirus is not uncommon in this community and that older children continue to be exposed to and become infected with rotavirus. PMID- 2556806 TI - Long term survival after pulmonary resection for small cell carcinoma of the lung. AB - A retrospective review was undertaken of the long term survival of 97 patients with histologically proved small cell carcinoma of the lung resected during the 10 years January 1977-December 1986. Twenty seven patients (28%) had stage I disease, 29 (30%) stage II, and 41 (42%) stage III. Patients with stage I and II tumours were managed by resection alone. Patients with stage III disease received adjuvant chemotherapy (cyclophosphamide, doxorubicin, and vincristine). Pneumonectomy was undertaken in 75 patients, lobectomy in 21, and wedge resection in one patient. Three patients died within 30 days of operation. The cumulative five year survival of all patients, irrespective of tumour stage, was 17%. The cumulative five year survival was 35% for patients with stage I disease, 23% for stage II, and zero for stage III. The median survival for patients with stage III tumours was 17 months. There was no significant difference in cumulative survival between patients with stage I and II disease. Cumulative survival, however, was significantly better for patients with stage I and II disease than for those with stage III disease. The data suggest that for patients with stage I and stage II disease surgery offers the prospect of long term survival. PMID- 2556807 TI - Dose-response study of nebulised nedocromil sodium in exercise induced asthma. AB - Ten patients with exercise induced asthma, in whom inhaled nedocromil sodium 4 mg by metered dose inhaler attenuated the exercise fall in forced expiratory volume in one second (FEV1) by at least 40%, participated in a double blind dose response study to compare the protective effect of nedocromil sodium given 15 minutes before exercise challenge via a nebuliser (Wright) in concentrations of 0.5, 5, 10, and 20 mg/ml with that of placebo (saline). Response was assessed as the maximum fall in FEV1 after the patient had run on a treadmill for six to eight minutes. Plasma concentrations of nedocromil sodium were measured at the time of challenge. After exercise challenge the mean (SEM) maximum percentage falls in FEV1 were 30.3 (1.6) for the control run and 28.0 (4.1) after placebo. The percentage fall was attenuated by pretreatment with all concentrations of nedocromil sodium to 12.8 (2.8), 11.2 (2.1), 12.8 (2.1), and 14.1 (3.5) for the 0.5, 5, 10, and 20 mg/ml concentrations respectively (p less than 0.001). There were no significant differences between the different nedocromil concentrations. Mean plasma concentrations of nedocromil were proportional to dose. Thus concentrations of nebulised nedocromil sodium that ranged from 0.5 to 20 mg/ml gave a similar degree of protection (50-60%) against exercise induced asthma. This appears to be the maximum protection that can be achieved with nedocromil sodium and is similar to the protection obtained with 4 mg nedocromil administered by metered dose aerosol. PMID- 2556808 TI - Chronic necrotising pulmonary aspergillosis treated with itraconazole. AB - A patient with longstanding ankylosing spondylitis developed chronic necrotising pulmonary aspergillosis, which was successfully treated with itraconazole. PMID- 2556810 TI - Ouabain enhances basal and stimulus-induced cytoplasmic calcium concentrations in platelets. AB - Incubation of platelet-rich plasma (PRP) with ouabain, an inhibitor of sodium/potassium ATPase (Na+/K+ ATPase), induced a significant rise in basal platelet intracellular calcium concentration [( Ca2+]i) when measured using fura 2. Ouabain induced an enhanced aggregation response to low doses of collagen in both PRP and washed platelets loaded with aequorin. In aequorin loaded platelets this enhanced aggregation response was associated with an enhanced rise in [Ca2+]i such that the relationship between [Ca2+]i and aggregation was unchanged. As inhibition of plasma membrane Na+/K+ ATPase would lead to a raised intracellular sodium ion concentration [( Na+]i) the results suggest that in the platelet, [Na+]i can modulate [Ca2+]i and hence influence the response of platelets to stimuli such as collagen. PMID- 2556809 TI - AIDS and the lung. 2--Antiretroviral treatment in human immunodeficiency virus disease. PMID- 2556811 TI - Protein C inhibitor from human plasma: characterization of native and cleaved inhibitor and demonstration of inhibitor complexes with plasma kallikrein. AB - Protein C inhibitor (PCI) was purified from human plasma using immunoaffinity chromatography and heparin Sepharose chromatography, a method that allowed the purification of active and inactive inhibitor. PCI purified from outdated plasma was inactive and either in complex with plasma kallikrein or proteolytically degraded. Sequence analysis of cleaved PCI and of complexes between PCI and activated protein C or urokinase identified the previously recognized inhibitor cleavage site Arg354-Ser355. Two additional cleavage sites were observed in the modified inhibitor i.e. Arg357-Leu358 and Arg362-Leu363 which probably represent secondary cleavage of the inhibitor. Furthermore the sequence analysis of the inhibitor, whether purified from fresh or outdated plasma, revealed that it was microheterogeneous in the NH2-terminus as a result of cleavage by a trypsin like enzyme(s). PMID- 2556813 TI - The relationship between anti-factor Xa level and clinical outcome in patients receiving enoxaparine low molecular weight heparin to prevent deep vein thrombosis after hip replacement. AB - Studies in experimental animals have demonstrated that there is a relationship between levels of low molecular weight (LMW) heparin and both bleeding and inhibition of thrombosis. The relationship between these outcomes and ex vivo anti-factor Xa levels has been examined in 163 patients undergoing total hip replacement who were given prophylaxis once daily with a LMW heparin (enoxaparine). Fifty patients received 60 mg of enoxaparine and 113 received 40 mg, both regimens being administered subcutaneously once daily. Blood samples for anti-factor Xa levels were collected 12 hours after the injection on the day of surgery and on days 1, 3 and 6, postoperatively. The incidence of wound hematoma was 5.3% when the maximum anti-factor Xa level was less than or equal to 0.2 units per ml, but increased to 24.5% when the anti-factor Xa level exceeded 0.2 units per ml, P = 0.0008. The incidence of postoperative thrombosis was low (6.3%) if the minimum anti-factor Xa level exceeded 0.1 units per ml, but increased to 14.6% when less than or equal to 0.1 units per ml, and to 18.8% if the anti-factor Xa level was less than or equal to 0.05 units per ml. Regression analysis revealed that there was a statistically significant relationship between anti-factor Xa level and wound hematoma, P = 0.002 and anti-factor Xa level and thrombosis, P = 0.03. These findings suggest that when enoxaparine is administered as a once daily subcutaneous injection, the 12 hour anti-factor Xa level should not exceed 0.2 units per ml to minimize bleeding and levels greater than 0.05 units per ml should be obtained to optimize efficacy. PMID- 2556812 TI - Effect of standard heparin and a low molecular weight heparin on thrombolytic and fibrinolytic activity of single-chain urokinase plasminogen activator in vitro. AB - The effect of unfractioned heparin (UH) and low molecular weight heparin (LMWH) (Kabi 2165 - Fragmin) on in vitro scu-PA thrombolytic and fibrinogenolytic activity was investigated. Thrombolytic activity was evaluated by following lysis of radiolabeled plasma clot immersed in plasma in presence of scu-PA alone or with either form of heparin. A 200 IU/ml scu-PA concentration produced clot lysis within 7 hr. UH or LMWH led to a slightly faster clot lysis which was statistically significant only at the 2nd and 3rd hour. No significant difference could be evidenced between UH and LMWH effect. During clot lysis, plasmin, generated within the clot led to a gradual transformation of scu-PA to tcu-PA, specially after a 4-hr incubation. Appearance of tcu-PA activity in the plasma surrounding the clot was significantly inhibited by either form of heparin. This finding contrasts with results observed in purified systems and suggests the presence of heparin-dependent plasma factor(s) inhibiting tcu-PA formation or its activity. Possible candidates might be anti-thrombin III and PAI-3. No fibrinogen breakdown was observed when plasma was incubated for 7 hr at 37 degrees C in presence of scu-PA alone (200 IU/ml) or with either form of heparin. However, in presence of a plasma clot, an important fibrinogen breakdown was observed during clot lysis reflecting the action of plasmin and/or tcu-PA generated within the clot, in the surrounding plasma. Fibrinogenolysis was less pronounced in the presence of both heparin preparations possibly as a consequence of the reduction in the tcu-PA level. These results underline the importance of plasma factors in the interaction of heparin with plasminogen activators such as scu-PA. PMID- 2556814 TI - The effect of calcium chloride on anti-Xa activity of heparin and its molecular weight fractions. AB - The anti-Xa activities of unfractionated heparin (UFH) and nine low molecular weight heparins (LMWH) have been measured in the presence and absence of 3 mM CaCl2, using bovine Factor Xa, purified human ATIII and an amidolytic assay. The addition of CaCl2 increased the activity of UFH by 93%, but the effect on LMWH was less, ranging from -20% to +55%. Studies of gel filtration fractions of UFH showed marked Mr dependence of the CaCl2 effect in the range 4,000-12,000. The differences among the various LMW heparins with respect to the effect of CaCl2 were closely correlated with the amount of polysaccharide above an Mr of 6,500. Kinetic studies confirmed the potentiation of the activity of UFH with bovine Xa and showed an even more marked effect using human Xa. PMID- 2556815 TI - Inhibition of platelet thromboxane formation and phosphoinositides breakdown by osthole from Angelica pubescens. AB - Osthole, isolated from Chinese herb Angelica pubescens, inhibited platelet aggregation and ATP release induced by ADP, arachidonic acid, PAF, collagen, ionophore A23187 and thrombin in washed rabbit platelets. It showed a weak activity in platelet-rich plasma. Osthole inhibited the thromboxane B2 formation caused by arachidonic acid, collagen, ionophore A23187 and thrombin in washed platelets, and also the thromboxane B2 formation caused by the incubation of lysed platelet homogenate with arachidonic acid. The generation of inositol phosphates in washed platelets caused by collagen, PAF and thrombin was suppressed by osthole. These data indicate that the inhibitory effect of osthole on platelet aggregation and release reaction was due to the inhibition of thromboxane formation and phosphoinositides breakdown. PMID- 2556816 TI - The ability of murine cytomegalovirus and class I major histocompatibility complex-disparate parental cells to induce alterations characteristic of severe graft-versus-host reactions. AB - The present studies were undertaken to examine the ability of a viral pathogen to enhance immune alterations associated with parent----F1 graft-versus-host reactions (GvHR) across defined donor/recipient MHC genetic disparities. Murine cytomegalovirus (MCMV) was administered concurrently with a parental lymphoid inoculum into unirradiated F1 recipients in strain combinations limiting allogeneic differences to the entire MHC complex (class I/II), the H-2K region (class I), or H-2IA (class II) regions only. Alterations previously found to be associated with GvHR involving changes in the expression of Ly-6, Lyt-2, and L3T4 were examined to characterize the effects of MCMV. Mice receiving low numbers of class I/II-disparate parental cells or MCMV alone failed to exhibit significant GvHR-associated changes. In contrast, introduction of cells and virus resulted in marked alterations characteristic of F1 recipients injected with a large parental cell inoculum alone. Concurrent virus and parental cells could also induce marked changes when administered across differences involving only a class I-disparate- but not class II-disparate only--P----F1 combination. In addition to the phenotypic changes observed during the concurrent virus and class I GvHR, markedly reduced spleen cell proliferative activity and associated weight loss and mortality appeared to indicate that virus had enhanced this reaction. In total, these findings demonstrated that a donor/recipient class I MHC difference was necessary for virus and parental cells to induce the changes observed, and thus not all donor/recipient antigenic differences will result in a similar virus induced effect. The results are discussed with respect to the potential mechanisms that may account for the apparent exacerbation of GvHR-associated alterations. PMID- 2556817 TI - Cytomegalovirus antigenemia as a useful marker of symptomatic cytomegalovirus infection after renal transplantation--a report of 130 consecutive patients. AB - In earlier work we demonstrated that CMV immediate early antigens can be detected in peripheral blood leukocytes of patients with active CMV infection. We now report a comparison of the antigenemia assay and an anti-CMV ELISA in a prospective longitudinal study of 130 renal transplant recipients who were monitored for active CMV infection during the first 3 months after transplantation. Active CMV infection developed in 56 patients. The antigenemia assay had a sensitivity of 89% and a specificity of 93% in the diagnosis of active CMV infection; for the ELISA these figures were 95 and 100%, respectively. In 22 of the 56 patients a CMV syndrome occurred. Antigenemia was demonstrated in all 22 patients while an antibody response occurred in 21 of them. The antigenemia assay became positive 8 +/- 7 days before the onset of symptoms while the antibody response was observed 4 +/- 9 days after the onset of symptoms. The pattern of antigenemia was helpful for monitoring the course of the infection. The maximum level of antigenemia was significantly higher and its duration significantly longer in symptomatic than asymptomatic infection. We conclude that CMV antigenemia is a sensitive, specific, and early marker of CMV infection. The antigenemia assay is of great value in monitoring patients with a high risk of CMV infection. PMID- 2556818 TI - Mussel poisoning and excitatory amino acid receptors. PMID- 2556819 TI - Steroid modulation reveals further complexity of GABAA receptors. PMID- 2556820 TI - Receptor-mediated Ca2+ entry: diversity of function and mechanism. PMID- 2556821 TI - The signal transduction system of the leukotriene D4 receptor. AB - During the past several years, substantial progress in understanding the receptors and signal transduction processes for peptidyl leukotrienes has been reported. Receptors have been identified and characterized, the major steps in the signal transduction pathway have been described, and the genetic and epigenetic regulatory processes have been characterized. Very recent studies have defined the mechanisms by which LTE4 acts as a partial agonist at the LTD4 receptor. The cloning of the genes for the proteins involved in the major steps of the signalling process has also been initiated. Stanley Crooke and co-authors summarize this recent progress and present their current notions about the LTD4 receptor signalling process. PMID- 2556822 TI - Receptors that inhibit phosphoinositide breakdown. AB - Calcium-mobilizing receptors are believed to activate phospholipase C. Joel Linden and Therese Mary Delahunty summarize recent reports which indicate that activation of some receptors that inhibit the accumulation of Ca2+ within cells - notably receptors for adenosine, dopamine and several other neurotransmitters - can inhibit phosphoinositide metabolism. Two types of mechanism may be involved in these responses. Many instances of receptor-mediated inhibition of phosphoinositide breakdown can be detected only after a period of several minutes and may be secondary to receptor-mediated events that lower [Ca2+]i or activate certain protein kinases. In other instances the activation of receptors rapidly (within seconds) inhibits phosphoinositide breakdown, possibly via the activation of guanine nucleotide binding proteins that either directly, or by a rapid indirect action, inhibit phospholipase C. Putative mechanisms for direct and indirect regulation of phospholipase C are discussed. PMID- 2556823 TI - Lateral mobility measurement of cell surface components: applications for molecular pharmacology. PMID- 2556824 TI - Breast cancer in West Indian women in Trinidad. AB - A retrospective review of breast cancer in West Indian women in Trinidad is presented. Breast cancer constituted 17% of all cancers diagnosed and represented 26% of all cancers in females. The usual presenting symptom was a painless lump. The right breast and the upper outer quadrant were the common site of cancer. 85% of the patients were multiparous with an average of 4 children and 92% of them breast fed. Most patients presented in stage I disease, but a significant number of cases were in an advanced stage of the disease. Infiltrating duct carcinoma was the most frequent histological diagnosis and lobular carcinoma was seen in 1% of the cases. PMID- 2556826 TI - [Enoxaparin (Clexane), a low molecular weight heparin in preventing thrombosis]. PMID- 2556825 TI - [The ultrastructural characteristics of the corticotropic cells of the rat adenohypophysis]. AB - Ultrastructural characteristics of corticotropic cells have been studied on serial semithin and ultrathin sections of the hypophysis from intact rats by the immunofluorescence and electron microscopic methods. It is shown that the population of corticotropic cells is heterogeneous and consists of degranulated, moderate granulated, hypergranulated cells and transitional forms among them. Ultrastructurally corticotropic cells correspond to Siperstein-Nakayama's ACTH cells. Kurosumi-Kobayashi "corticotrophs" fail to react with the antiserum to ACTH, so they have not regard for its production. PMID- 2556827 TI - [Criteria for the early diagnosis of chronic kidney failure]. AB - Comprehensive study of blood and urinary levels of beta 2-microglobulin and cAMP in patients with chronic renal failure (CRF) demonstrated their significant alterations in the disease development. Therefore the levels of beta 2 microglobulin and cAMP could be of great aid in reaching the early diagnosis of CRF. PMID- 2556828 TI - Orchiectomy only for clinical stage I nonseminomatous germ cell testis tumors: comparison with pathologic stage I disease. AB - Twenty-three men with nonseminomatous germ cell tumors confined within the tunica albuginea of the testis and negative metastatic evaluation were placed on surveillance after radical orchiectomy. Metastatic disease developed in 3 for a relapse rate of 13 percent. During the same period, 17 men with pathologic Stage I disease were followed up in the same manner, and metastatic disease developed in 3 for a relapse rate of 18 percent. All six men were treated with either combination chemotherapy or retroperitoneal lymphadenectomy, and all are alive without disease for at least two years from end of therapy. If strict criteria are followed, surveillance has a place in the management of carefully selected young men with clinical Stage I disease. PMID- 2556829 TI - Correlation of clonogenic growth with histologic diagnosis after resection of residual tissue following chemotherapy for nonseminomatous testis cancer. AB - Patients who undergo chemotherapy for metastatic nonseminomatous germ cell cancer of the testis often undergo subsequent resection of residual tissue. We performed a retrospective review of 11 consecutive patients who had their resected residual tissue cultured in a soft agar clonogenic assay to determine if there were any biological data that could be obtained in this setting. Twelve assays were performed on the 11 patients. Some colony formation occurred in 10 of the 12 assays, including 4 of 5 assays in which cancer was found and 6 of 7 in which no cancer was found. The rate of colony formation, however, was significantly less for the noncancerous tissues than for the ones with cancer noted (p = 0.019). Three of the patients with cancer are dead, while all of those with benign tissue remain free of disease, with follow-up ranging from three to six years. Our data suggest that clonogenic growth from resected tissue after chemotherapy has the potential to supplement histologic findings, possibly as a predictor of the future biologic behavior of a patient's disease process. PMID- 2556830 TI - [Nephroblastoma of the right kidney simulating clinical picture of acute destructive cholecystitis]. PMID- 2556831 TI - Parainfluenza 3 vaccination of sheep. AB - Outbreaks of pneumonia associated with Pasteurella haemolytica have occurred in sheep in the area of Perthshire served by this practice, and on some farms the disease has been an important annual cause of loss. Serological evidence was obtained that parainfluenza 3 (PI3) virus might be implicated as a predisposing factor to pasteurellosis. A live attenuated PI3 virus vaccine licensed for use in cattle was given intranasally to ewes on one farm. Many sheep seroconverted and outbreaks of pneumonia were negligible around the subsequent lambing time. The protection of the flock appeared to last for one season only. Subsequently ewes and lambs on other farms were vaccinated and on these farms there were fewer deaths than expected due to pasteurellosis. PMID- 2556832 TI - Cowpox in cats. PMID- 2556833 TI - [Magnetic resonance tomography in the diagnosis of juvenile nasopharyngeal angiofibromas]. AB - This work is based upon the experience in the diagnosis of juvenile nasopharyngeal angiofibromas (JAF) in 14 patients using MR-tomography and routine x-ray examination. In all the patients MR-tomography made it possible to assess the topical position of a tumor, its spreading to the cranial, nasal, accessory sinusal and orbital cavities, and the pterygopalatine and infratemporal spaces. MR-tomographic diagnosis made it possible to differentiate between tumor invasion and inflammatory processes detecting JAF attending tubotitides. However MR tomography cannot be used alone to assess the status of osseous tissue. PMID- 2556834 TI - Nerve sheath myxoma of the breast. A light and electron microscopic, histochemical and immunohistochemical study. AB - A nerve sheath myxoma involving the breast has been examined by light and electron microscopy, and by immunohistochemical and histochemical methods. Electron microscopically, cells with features indicative of Schwann cells, perineural cells and fibroblasts were identified in the tumour and S-100 protein and vimentin positivity was demonstrated in the tumour cells. The mucoid matrix stained positive for chondroitin-4 or 6-sulphate in keeping with a cartilaginous lesion. These findings are discussed in relation to the uncertain histogenesis and the differential diagnoses of the tumour in the breast. PMID- 2556835 TI - [The effects of complex platinum compounds on the neuraminidase activity of the Sendai virus]. AB - The effect of di- and tetravalent cis-diaminoplatinum chlorides on Sendai virus envelop HN glycoprotein was investigated. The partial inhibition of neuraminidase activity was greater in the case of the divalent platinum complex derivative. PMID- 2556836 TI - Some physico-chemical properties of the rigid form of the Sendai virus nucleocapsid. AB - The effect of some dissociation agents (SDS, beta-mercaptoethanol, urea, EDTA) on the rigid form of the Sendai virus nucleocapsid was studied. Polyacrylamide gel electrophoresis in the presence of lytic mixture (1% SDS, 2% beta mercaptoethanol, 5 M urea, for 2 min at 100 degrees C) revealed two types of polypeptide subunits (mol. wts. 46,000 and 14,000), as well as the dissociation in the presence of 0.1% SDS only. The EDTA treatment leads to a disorganization of the protein part (10(-2) M) or of the nucleocapsid structure (5 x 10(-2) M). PMID- 2556837 TI - [Herpes simplex viral latency in human embryo fibroblasts treated with acyclovir]. PMID- 2556838 TI - Herpes simplex virus Vmw65-octamer binding protein interaction: a paradigm for combinatorial control of transcription. AB - The transcriptional status of a given viral or cellular gene is determined both by the availability of functional transcription factors and by the combination and spatial arrangement of the cis-acting elements to which they bind. While differential gene expression can be achieved to some extent by the interaction of different factors with different genes it is clear that in some cases the same factor is required for the regulation of genes that are not coordinately expressed. How a transcription factor achieves selective function in the absence of selective binding is a question that remains largely unanswered. The recent advances in understanding how both protein-protein and protein-DNA interactions are required to mediate the induction of herpes simplex virus immediate early (HSV IE) gene expression have highlighted a novel mechanism for combinatorial control of transcription which has significant implications for the differential control of cellular gene expression. PMID- 2556839 TI - Avian retrovirus integration protein: structure-functional analysis of viable mutants. AB - A replication-competent avian retrovirus mutant, containing a single amino acid substitution at amino acid residue 115 in the 3' endonuclease (IN) region of the polymerase (pol) gene, was characterized. DNA transfection experiments demonstrated that the mutant virus exhibited a delayed growth phenotype at 41 degrees while replicating efficiently at 35 degrees. Examination of virus infected cells at the molecular level demonstrated that the mutant virus at either temperature was capable of synthesizing viral DNA as efficiently as wild type Rous sarcoma virus, strain Prague A. This result suggested that the same mutation, which was also present in the IN moeity of the polymerase beta polypeptide, did not affect DNA synthesis. Further analyses demonstrated that at either temperature the mutant virus integrated its DNA at about 10-20% of wild type level, although possibly less efficiently at 41 degrees than at 35 degrees. The mutation at residue 115 (Pro to Ser) appeared to lower the ability of IN to function in the integration of viral DNA relative to wild-type virus. No definitive conclusion could be made as to whether IN in this mutant possessed a temperature-sensitive lesion which caused the observed replication defect at 41 degrees. PMID- 2556840 TI - Size and stability of the Epstein-Barr virus major internal repeat (IR-1) in Burkitt's lymphoma and lymphoblastoid cell lines. AB - We have used field inversion gel electrophoresis to survey EBV strains for the size of the major internal repeat, IR-1, and estimate the number of 3.1-kb repeat units present. The B95-8 strain of EBV was estimated to contain 8.6 repeats. The repeat number varies considerably among naturally occurring isolates around a mean of six repeats. Some cell lines harbored multiple viral genomes with differing numbers of repeats and our results suggest that the repeat number in IR 1 is more likely to change during lytic replication than during latency. The Jijoye strain had 6.6 repeats and the Jijoye deletion mutant clone P3HR-1 retained 5.9 repeats setting the size of the P3HR-1 deletion at 6.8 kb. Thus, the nonimmortalizing mutant has retained all of the W1 and W2 exons of the immortalizing parent and has lost only the 3' unique exons of EBNA4 and all of EBNA2. PMID- 2556841 TI - Interference with vaccinia virus growth caused by insertion of the coding sequence for poliovirus protease 2A. AB - Attempts were made to express noninfectious derivatives of full-length type 1 (Mahoney) and type 2 (Lansing) poliovirus cDNAs in live recombinant vaccinia viruses for vaccine purposes. Vaccinia virus (VV) would not tolerate insertions of polio cDNA containing the coding sequence for the polio protease 2A. However, polio cDNA with the 2A gene deleted either in vivo or in vitro could be inserted into VV and stably maintained. Genetic evidence indicated that expression of the polio 2A gene in trans from transfected plasmid DNA was deleterious to vaccinia virus within the same cell. The 2A product presumably interferes with VV growth by modifying the host translational machinery such that translation of host and vaccinia capped mRNAs is inhibited. Polio cDNA containing a mutated 2A gene whose product is no longer active in host protein shutoff could be inserted into VV. However, inserts containing the intact mutated 2A gene did not synthesize detectable poliovirus protein, although they did produce polio-specific RNA. Expression of polio-specific protein was detected from a VV-polio recombinant containing cDNA encoding the capsid proteins plus an incomplete 2A gene. These results have implications regarding possible vaccine construction, and suggest a mechanism for interference between polio and vaccinia viruses in mixed infection. PMID- 2556842 TI - The primary structure of major viral RNA in a rat cell line transfected with type 47 human papillomavirus DNA and the transforming activity of its cDNA and E6 gene. AB - A transformed cell line (RC335) showing higher saturation cell density was obtained from 3Y1 cells (a fibroblastic cell line of rat) transfected with DNA of human papillomavirus type 47 (HPV-47), an epidermodysplasia verruciformis associated virus, which our laboratory reported previously. The cell line was found to produce a major and several minor species of viral RNAs. The primary structure of the major viral RNA in RC335 was extensively studied and found to consist of two exons and contain open reading frames (ORFs) E6, E7, and a fused ORF, E1/E4. The major RNA was indicated to play an important role in the transformation of RC335 by an experiment with the recombinant retrovirus designed to produce the RNA containing these ORFs, i.e., the recombinant virus induced transformation similar to that in RC335 upon infection of 3Y1 cells. Furthermore the experiments with recombinant viruses carrying a nonsense mutation or large deletion in the above ORF(s) indicated that E6 was necessary and sufficient for the transformation. PMID- 2556843 TI - Physical structure and molecular cloning of equine cytomegalovirus DNA. AB - Restriction endonuclease (RE) mapping studies and molecular hybridization analyses were conducted to determine the molecular structure of the genome of equine cytomegalovirus (ECMV). The ECMV genome is a linear, double-stranded DNA with a molecular size of 126 +/- 0.6 MDa (189 kbp). A library of cloned BamHI, EcoRI, and HindIII fragments of the viral genome was used to construct RE maps. Individual 32P-labeled cloned DNA fragments were hybridized to Southern blots of viral genomic DNA digested to completion with BamHI, EcoRI, HindIII, or SalI. These analyses revealed that the ECMV genome consists of a 97-MDa unique long region which is bracketed by repeated sequences. At one terminus of the genome, a 21.3-MDa segment of repeated sequences with no apparent unique sequences was identified. At the other terminus, a 6-MDa unique region bracketed by 2.4-MDa repeat segments was identified. No submolar RE fragments were identified upon digestion of the ECMV genome with BamHI, EcoRI, HindIII, SalI, or other REs, including BclI, BglII, NruI, and XbaI. The genome possesses only two termini as judged by lambda exonuclease digestion and by T4 DNA polymerase end-labeling of the intact DNA followed by digestion with BamHI, EcoRI, HindIII, SalI, BclI, BglII, NruI, or XbaI. In addition, Southern blot analysis of DNA extracted from ECMV-infected rabbit kidney cells revealed that only one viral DNA fragment within the intracellular viral DNA pool contains fused genomic termini. Taken together, these observations indicate that the ECMV genome does not isomerize and suggest that the genome of ECMV may be unique among those of the herpesviruses and especially those of the betaherpesviruses (cytomegaloviruses) since it contains regions of extensive internal homology yet does not undergo isomerization. Lastly, the relatively small size of the viral genome indicates an evolutionary diversification among the cytomegaloviruses. PMID- 2556845 TI - Synthesis and processing of equine herpesvirus type 1 glycoprotein 14. AB - Glycoprotein 14 (gp14) of equine herpesvirus type 1 (EHV-1), the homolog of herpes simplex virus (HSV) glycoprotein B (gB), was investigated employing a panel of monoclonal antibodies to ascertain the regulatory class, rate of synthesis, and type of glycosylation of this polypeptide. Application of immunoprecipitation, Western blot, and SDS-PAGE analysis in conjunction with the use of metabolic inhibitors (cycloheximide, antinomycin D, phosphonoacetic acid, tunicamycin, and monensin), and time-course and pulse-chase experiments revealed the following information: (1) Three gp14-related polypeptides with molecular weights of 138 kilodaltons (K), 77-75K, and 55-53K are present in EHV-1-infected cell extracts. (2) All three species are synthesized in the presence of the DNA synthesis inhibitor phosphonoacetic acid although their synthesis is enhanced by DNA replication, indicative of a beta-gamma class molecule. (3) The 138K species is synthesized first as a precursor of the smaller species of gp14, the 77-75K and 55-53K forms. (4) Use of glycosylation inhibitors and digestion of immunoprecipitated gp14 with endoglycosidases indicate that the primary translation product is a 118K molecule which is cotranslationally glycosylated to the 138K form by the addition of high mannose oligosaccharides. (5) The 77-75K species contains both high mannose and hybrid oligosaccharides while the 55-53K form of gp14 contains some complex oligosaccharides. (6) In the absence of a reducing agent, the 138K polypeptide and a large 145K species are observed in both infected cell extracts and purified virions. Thus, EHV-1 gp14 appears to be synthesized as a large precursor molecule of 138K and is proteolytically cleaved to two smaller forms, 77-75K and 55-53K, which are linked by a disulfide bond(s) to form a 145K complex. This model of gp14 synthesis and maturation is similar to those proposed for a number of HSV gB equivalents found in the Alphaherpesvirnae. PMID- 2556844 TI - Expression of the peplomer glycoprotein of murine coronavirus JHM using a baculovirus vector. AB - The gene encoding the E2 peplomer glycoprotein of coronavirus mouse hepatitis virus JHM strain (JHMV) has been inserted into the genome of Autographa californica nuclear polyhedrosis baculovirus (AcNPV) in lieu of the coding region of the AcNPV polyhedrin gene. This recombinant virus produced E2 protein in insect cells under the control of the baculovirus polyhedrin promotor. The expressed E2 protein was shown in size and antigenic properties to be similar to the E2 protein produced in mouse cells infected by JHMV. The expressed E2 protein was glycosylated and transported to the cell surface; however, no proteolytic cleavage was detected in insect cells. The sera from rats immunized with partially purified E2 protein derived from insect cells reacted in immunoprecipitation and immunofluorescence experiments with the E2 protein produced in JHMV-infected mouse cells. The antiserum failed to neutralize the infectivity of JHMV. These results suggest that the E2 protein expressed by the recombinant baculovirus in insect cells is similar but not identical to the E2 protein produced in JHMV-infected mouse cells. The inability of the E2 protein expressed in insect cells to produce neutralizing antibody is discussed. PMID- 2556846 TI - Sequence analysis reveals extensive polymorphism and evidence of deletions within the E2 glycoprotein gene of several strains of murine hepatitis virus. AB - Direct RNA sequence analysis of the E2 gene of wild-type MHV-4 and of neutralization resistant, neuroattenuated variants has identified a polymorphic region with respect to deletions. These variants had large deletions of 142 to 159 amino acids mapping to a localized region in the amino-terminal domain of the peplomer glycoprotein. The nucleotide sequence of the E2 gene for wild-type strain MHV-4 was found to be very similar to that of MHV-JHM but had an insertion of 423 nucleotides resulting in the addition of a stretch of 141 unique amino acids in the amino-terminal domain of E2. We propose that deletions reflect a major source of heterogeneity in the E2 protein of MHV. PMID- 2556848 TI - Mapping of two varicella-zoster virus-encoded genes that activate the expression of viral early and late genes. AB - A transient assay system was used to identify varicella-zoster virus (VZV) encoded genes whose products are able to activate the expression of an early gene promoter, the thymidine kinase (tk) promoter, and a late gene promoter, and the glycoprotein I (gpI) promoter. Vero cells were cotransfected with individual cloned DNA fragments spanning the entire VZV genome and with the recombinant construct p1tkCAT which contained the chloramphenicol acetyl transferase (CAT) gene under the control of putative regulatory sequences. Five- to 20-fold increases in the expression p1tkCAT was observed in cotransfections with plasmids containing VZV open reading frame (ORF)4 (map location 0.02-0.03) or ORF62 (0.82 0.86). Expression of p68CAT (contains -682 to +222 bp relative to the AUG of gpI) was also enhanced by the products of ORF4 and ORF62. Synergy between ORF4 and ORF62 products was observed in the activation of p68CAT, resulting in a 22-fold increase in CAT activity. RNA analysis indicated that activation of these promoters was at the transcriptional level. A VZV-encoded "repressor" sequence, containing ORF60 and ORF61, was also identified which repressed expression of p1tkCAT and modulated its activation by ORF4 and ORF62. PMID- 2556847 TI - Biosynthesis, structure, and biological activities of envelope protein gp65 of murine coronavirus. AB - We have previously shown that gp65 (E3) is a virion structural protein which varies widely in quantity among different strains of mouse hepatitis virus (MHV). In this study, the biosynthetic pathway and possible biological activities of this protein were examined. The glycosylation of gp65 in virus-infected cells was inhibited by tunicamycin but not by monensin, suggesting that it contains an N glycosidic linkage. Glycosylation is cotranslational and appears to be complete before the glycoprotein reaches the Golgi complex. Pulse-chase experiments showed that this protein decreased in size after 30 min of chase, suggesting that the carbohydrate chains of gp65 undergo trimming during its transport across the Golgi. This interpretation is supported by the endoglycosidase treatment of gp65, which showed that the peptide backbone of gp65 did not decrease in size after pulse-chase periods. This maturation pathway is distinct from that of the E1 or E2 glycoproteins. Partial endoglycosidase treatment indicated that gp65 contains 9 to 10 carbohydrate side chains; thus, almost all of the potential glycosylation sites of gp65 were glycosylated. In vitro translation studies coupled with protease digestion suggest that gp65 is an integral membrane protein. The presence of gp65 in the virion is correlated with the presence of an acetylesterase activity. No hemagglutinin activity was detected. PMID- 2556849 TI - Differential regulation by varicella-zoster virus (VZV) and herpes simplex virus type-1 trans-activating genes. AB - Transient expression assays were performed in Vero cells in order to compare varicella-zoster virus (VZV)-encoded trans-activating proteins [defined by the products of open reading frames (ORF) 4 and 62] with herpes simplex virus type-1 (HSV-1) trans-activating proteins, ICP4 and ICP0, with respect to activation of gene expression. We demonstrate that the product of VZV ORF4 and ORF62 (which are the HSV-1 analogs of ICP27 and ICP4, respectively) stimulate a variety of viral and cellular gene promoters, including the HSV-1 thymidine kinase (tk) promoter. On the other hand, expression of a recombinant vector containing the VZV tk promoter could not be stimulated, by HSV-1 infection or by the HSV-1 ICP4 or ICP0 proteins expressed during cotransfection experiments. These data suggest different mechanisms of activation of the VZV and the HSV-1 tk gene promoters by "trans-activating" factors. PMID- 2556850 TI - Herpes simplex virus-induced "rolling circle" amplification of SV40 DNA sequences in a transformed hamster cell line correlates with tandem integration of the SV40 genome. AB - Infection with herpes simplex virus leads to amplification of SV40 DNA in various SV40-transformed cells. In earlier studies with the SV40-transformed hamster cell line Elona two different types of DNA amplification could be identified: (i) Bidirectional overreplication of chromosomally integrated SV40 DNA expanding into the flanking cellular sequences ("onion skin" type) and (ii) highly efficient synthesis of extremely large head-to-tail concatemers containing exclusively SV40 DNA ("rolling circle" type). These investigations have indicated that the chromosomally integrated form of SV40 might be the substrate for both types of overreplication. There still had been uncertainties as to whether and how these events were connected. A hypothetical assumption of a recombinational event leading to the excision of SV40 DNA molecules is supported by the results presented here: In this study cloned Elona cell lines were investigated for their ability to amplify SV40 sequences and for the mechanism of amplification utilized. SV40 integration in a partial tandem manner correlates with a strong rolling circle amplification. In contrast, in one cell line harboring a truncated SV40 genome, amplification appears mainly restricted to intrachromosomal bidirectional overreplication. Possible implications for HSV functions involved in the amplification process will be discussed. PMID- 2556851 TI - Herpes simplex virus type 1 ribonucleotide reductase null mutants induce lesions in guinea pigs. AB - Two herpes simplex virus type 1 ribonucleotide reductase null mutants, hrR3 and ICP6 delta, produced cutaneous lesions in guinea pigs as severe as those of wild type strains. The lesions induced by hrR3 resulted from in vivo replication of the mutant virus, suggesting that this virus-encoded enzyme is nonessential for virus replication in guinea pigs. PMID- 2556852 TI - Chromatographic analysis of the aminoacyl-tRNAs which are required for translation of codons at and around the ribosomal frameshift sites of HIV, HTLV 1, and BLV. AB - An examination of the frameshift signals or proposed signals within published sequences of retroviruses and other genetic elements from higher animals shows that each site utilizes a tRNA which normally contains Wybutoxine (Wye) base or Queuine (Q) base in the anticodon loop. We find experimentally that most of the Phe-tRNA present in HIV-1 infected cells lacks the highly modified Wye base in its anticodon loop and most of the Asn-tRNA in HTLV-1 and BLV infected cells lacks the highly modified Q base in its anticodon loop. Interestingly, Phe-tRNA translates a UUU codon within the ribosomal frameshift signal in HIV and Asn-tRNA translates a AAC codon within the proposed frameshift signals in HTLV-1 and BLV. Thus, the lack of a highly modified base in the anticodon loop of tRNAs in retroviral infected cells is correlated with the participation of these undermodified tRNAs in the corresponding frameshift event. This suggests that the "shifty" tRNAs proposed by Jacks et al. (Cell 55, 447-458, 1988) to carry out frameshifting may be hypomodified isoacceptors. PMID- 2556853 TI - Nucleotide sequence of gene segment 1 of a porcine rotavirus strain. AB - The nucleotide sequence of gene segment 1, which encodes VP1 of porcine rotavirus strain Gottfried, was determined. VP1 is associated with single-shelled rotavirus particles and has been linked to virus transcriptase and replicase enzymatic activities. Gene segment 1 is 3302 nucleotides long with a single open reading frame capable of coding for a protein of 1088 amino acids (calculated mol wt 125 kDa). The predicted amino acid sequence revealed that VP1 is basic, with a net positive charge of 18 at pH 7.0. It shares five consensus sequences with several well-characterized RNA-dependent RNA polymerases. Gottfried VP1 also shares consensus sequences with certain GTP-binding proteins; however, we could not detect any GTP-binding activity in VP1. Our preliminary experiments suggest that VP3, another polypeptide located in single-shelled rotavirus particles, possesses GTP-binding activity. These results suggest that mRNA synthesis and capping enzyme activities are related to VP1 and VP3, respectively. PMID- 2556854 TI - Disparate effects of two herpesvirus [corrected] immediate-early gene trans activators on the HIV-1 LTR. AB - The BMLF1 region of the Epstein-Barr virus (EBV) genome and the immediate-early (IE) region of human cytomegalovirus (HCMV) both encode proteins which can trans activate heterologous promoter/chloramphenicol acetyl transferase (CAT) constructs, including a human immunodeficiency virus type-1 promoter/CAT construct. We demonstrate here that this trans-activation by the EBV BMLF1 gene product, which we have previously shown to be largely post-transcriptional, is reporter gene dependent. In contrast, trans-activation by the HCMV-IE gene product(s), previously shown to be mediated at the RNA level, is seen regardless of whether CAT, human growth hormone, or beta-galactosidase is used as the reporter gene. Mutational analysis revealed no specific cis-acting sequences within the HIV-1 promoter which were required for trans-activation by the HCMV-IE gene product(s). PMID- 2556855 TI - Glucocorticoid-enhanced neoplastic transformation of human keratinocytes by human papillomavirus type 16 and an activated ras oncogene. AB - Human papillomaviruses (HPV) are known etiological agents of benign proliferation of the skin and mucosa (papillomas and warts). They have also been implicated in the development of cervical dysplasia and anogenital carcinoma. The close association of HPV type 16 DNA with the majority of cervical carcinomas suggests the involvement of the virus in this type of cancer. We have developed an in vitro multistep model for human epithelial cell carcinogenesis. Primary human epidermal keratinocytes acquired an indefinite lifespan in response to transfection with HPV 16 DNA but did not undergo malignant conversion. Addition of Kirsten murine sarcoma virus (Ki-MSV), which contains an activated K-ras oncogene, to these cells induced morphological alterations associated with the acquisition of neoplastic properties. The frequency of transformation by Ki-MSV was markedly enhanced by the inclusion of glucocorticoid. At optimal conditions, a 125-fold stimulation was observed. These findings demonstrate the malignant conversion of human primary epithelial cells in culture by the cooperation of HPV type 16 and an activated ras oncogene and support a multistep process for neoplastic conversion. The availability of a human epithelial cell transformation model should facilitate studies of the interaction between HPV and human epithelial cells. PMID- 2556856 TI - Estrogen regulation of protein synthesis and cell growth in human breast cancer. PMID- 2556857 TI - Pyrroloquinoline quinone: a novel cofactor. PMID- 2556858 TI - [The possibility of using an indirect method of fluorescent antibodies to the hepatitis A virus in epidemic control practice]. PMID- 2556859 TI - [Protein S-100 in spontaneous soft tissue and peripheral nerve neoplasms in rats]. AB - 16 spontaneous tumors of the peripheral nerves and 18 spontaneous tumors of mesenchymal origin in BDVI rats were studied by peroxidase-antiperoxidase method using anti-serum (DAKOPATT) against protein S-100. The majority of spontaneous peripheral nerve tumors were of cystic histological structure identical to that of cystic neurinomas induced in rats by ethylnitrosourea and almost all of these tumors were S-100 protein positive. The incidence of spontaneous neurinomas in BDVI rats was in some experiments as high as 5%. All tumors of mesenchymal origin (except one lipoma) were S-100 protein negative: 2 fibromas, 6 fibrosarcomas, 3 malignant fibrous histiocytomas, one rhabdomyosarcoma and one hemangioendothelioma. S-100 protein is found, as in human pathology, useful for distinguishing tumors of schwann cell and mesenchymal origin in rats. PMID- 2556860 TI - [Tactics in treating patients with small cell cancer of the lung with superior vena cava obstruction syndrome]. AB - A non-randomized study was carried out to evaluate the immediate and end results of chemo-radiation therapy of 44 cases of small-cell lung cancer aggravated with superior vena cava obstruction syndrome. Both radiation and chemotherapy proved effective in urgent combatting said syndrome. The best results were reliably secured when chemotherapy was supported with radiotherapy at an earlier possible stage. Due to gravity of their condition such case require the therapy as an urgent measure. The treatment proved effective as 40% of patients with localized small-cell lung tumors and the syndrome survived 12 months. PMID- 2556862 TI - [Dermatomycoses and an antifungal diet]. AB - Treatment of dermatomycoses and some other dermatoses, e.g. seborrheic eczema, is more effective if the intestinal tract is free of pathogenic fungi. In order to reduce the yeast colonies between the intestinal villi and to avoid the persorption of living yeast cells into the lymph-tract and the blood circulation it is recommendable to perform the so-called antifungal diet: less sugar of all kind and much more vegetable fibres in the food. PMID- 2556861 TI - [Interaction studies with fluconazole, a new triazole antifungal drug]. AB - Fluconazole is a new systemic antifungal agent from the class of triazoles. Its efficacy was documented in clinical studies of patients with candidosis and cryptococcosis. Due to the influence on human cytochrome-P-450-dependent enzymes, special attention should be paid, in the development of new antifungal agents, to the inhibition of the metabolization of other simultaneously given substances and to the inhibition of steroid hormone synthesis in the gonads and adrenal glands. The interaction studies carried out with frequently administered drugs such as oral contraceptives, cimetidine, warfarin, tolbutamide, and cyclosporin (transplantation patients) allow the concomitant intake of these substances with fluconazole. Concomitant anti-coagulant treatment requires a precautionary monitoring of prothrombin time; it may be necessary in individual cases to reduce the anticoagulant dosage. The intake of oral hypoglycemics of the tolbutamide type requires monitoring of blood glucose, although blood glucose lowering effects were not found. One interaction study carried out with phenazone showed the low metabolization tendency of fluconazole. Approximately 80% of the administered fluconazole dose appeared in the urine unchanged. Studies in male and female subjects with multiple doses of fluconazole (4 weeks) did not show any significant effects on the endocrinology and steroid synthesis in man. PMID- 2556863 TI - Borderline ovarian tumors: a review of treatment. AB - Borderline ovarian tumors have an excellent prognosis. In stage I disease, no therapy in addition to surgery is needed, and conservation of ovarian tissue for future childbearing may be appropriate. In advanced stages, the use of adjuvant therapy has not consistently led to cures, and complications have been reported. A randomized study of no adjuvant therapy versus adjuvant treatment with long term follow-up will be necessary to determine the efficacy of additional treatment. PMID- 2556865 TI - [The spine as an unusual location for a malignant synovioma]. AB - The authors report on a case of malignant synovioma of the spine, a rare condition. A general review of the literature on the pathogenesis and localization, and the histologic and immunohistochemical investigation possibilities for differential diagnosis is given. The surgical and radiotherapeutic treatment possibilities are pointed out. PMID- 2556866 TI - [The physiology of the blood vessels (research results and prospects)]. PMID- 2556864 TI - Oncogene expression in vivo by ovarian adenocarcinomas and mixed-mullerian tumors. AB - Six-micron paraffin sections of paraformaldehyde-fixed specimens of 24 ovarian benign and neoplastic specimens were assayed for tumor cell-specific oncogene expression by a sensitive, quantitative in situ hybridization technique with probes for 17 oncogenes, beta-actin, and E. coli beta-lactamase. In the benign, borderline, and invasive adenocarcinomas, multiple oncogenes, including neu, fes, fms, Ha-ras, trk, c-myc, fos, and PDGF-A chains, were expressed at significant levels relative to a housekeeping gene (beta-actin). In the mixed-Mullerian tumors, a rather different pattern of oncogene expression was observed, characterized primarily by expression of sis (PDGF-B chain). For the adenocarcinomas, statistical analysis demonstrated that expression of several genes (fms, neu, PDGF-A) was closely linked to others (c-fos, c-myc) known to have important roles in the control of cell proliferation, but only one gene, fms, correlated very strongly with clinicopathologic features (high FIGO histologic grade and high FIGO clinical stage) predictive of aggressive clinical behavior and poor outcome. The authors discuss the role that tumor epithelial cell expression of the fms gene product might play in the auto- and paracrine control of growth and dissemination of ovarian adenocarcinomas. PMID- 2556867 TI - [End-plate currents and characteristics of the neuromuscular synapse]. PMID- 2556868 TI - [Nephroblastomas (Wilms' tumors) and special variations of nephroblastomas]. AB - The results of the National Wilms' Tumor Study (NWTS) enabled the subdivision of nephroblastomas into subtypes with "favorable and unfavorable histology". Nephroblastomas with "unfavorable histology" could be discriminated by identifying those tumors not responding to therapeutic regimes proven successful for most cases with "favorable histology". A major disadvantage of the NWTS classification has been the exclusion of cytodifferentiated nephroblastoma variants, which, in contrast to typical nephroblastomas, can be cured by complete nephrectomy with wide excision of perinephric soft tissue. In the current study all types of nephroblastoma and nephroblastoma variants were included to encompass the whole morphological spectrum which these tumors may assume. This unselected material is necessary to define the relation between morphology and prognosis and to compare the treatment results of various clinical trials. Three hundred and four cases of nephroblastoma and related neoplasms on file at the Pediatric Tumor Registry, Kiel, were investigated by conventional light microscopy, electron microscopy, immunohistochemistry and DNA-flow cytometry. Of the "typical" nephroblastomas 50% occurred in the left kidney, 45% in the right kidney, and 5% were bilateral. Five cases were located in extrarenal sites. There were 121 males and 114 females. The peak incidence was noted in the third year of life. Of 135 patients 111 are alive and well, nine are living with disease, and 10 patients have died of disease. The blastemal predominant and stromal predominant types in our study were more frequent than in the NWTS. By contrast, the mixed and epithelial predominant types were more frequent in the NWTS. Patients with nephroblastomas of mixed or blastemal predominant type were older than those with epithelial predominant or stromal predominant type. Electron microscopy showed that nephroblastoma is derived from metanephric blastema. Blastemal cells are capable of differentiating into tubular epithelial cells and stromal cells. Undifferentiated blastemal cells contain exclusively vimentin intermediate filaments, better differentiated blastemal cells vimentin and cytokeratin, and stromal cells exclusively vimentin. Preoperative radio- and/or chemotherapy led to a marked reduction of undifferentiated blastema and poorly differentiated tubules, whereas better differentiated tubules, striated muscle, hyaline cartilage, cells with anaplastic and sarcomatous elements were not affected. Thus, identification of highly malignant nephroblastomas with anaplasia and sarcomatous renal tumors was even possible after preoperative treatment. Congenital mesoblastic nephroma (CMN; n = 17) is a low-grade malignant, cytodifferentiated nephroblastoma which very rarely occurs beyond the fourth month of life and has an excellent prognosis, provided it has been completely resected.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2556869 TI - [Unexpected effects of a whole body irradiation on the mortality rate of baby mice after an experimental infection with the vesicular stomatitis virus (VSV)]. AB - The effect of total body irradiation of baby mice or their pregnant mothers with surface doses of 3.9 Gy on the mortality rate after infection of the baby mice with VSV has been studied. Irradiation of the newborns had no influence on the mortality rate of the infected animals. Irradiation of the pregnant animals--and the foetuses in utero--caused an astonishing decrease of the mortality rate of the infected baby mice for about 50% in compare to the non irradiated controls. PMID- 2556870 TI - Rapid diagnosis of TGEV-like coronavirus in fattened pigs by indirect immunofluorescence labelling in nasal cells. AB - Diagnosis of TGEV-like coronavirus was made by an immunofluorescence technique using monoclonal antibodies directed against TGEV. Six, 15 weeks-old pigs were inoculated intratracheally with the strain P 6008 ST 4. No clinical sign was observed during the course of the infection. Viral antigens were detected in the cytoplasm of nasal cells obtained from swab in five animals as early as one day p.i. and during several days. The sensitivity and specificity of this method appeared to be comparable to virus isolation of this cell-adapted TGEV-like strain. PMID- 2556871 TI - [Isolation of coronaviruses in cell culture from nasal swab samples of calves with respiratory tract diseases in the Federal Republic of Germany]. AB - Coronavirus was isolated in HRT-18 tissue culture from nasal swabs of 4 from 15 calves showing respiratory symptoms. The strains showed cytopathic effects and could be identified as coronaviruses of bovine origin by their mouse erythrocyte agglutinating activity as well as serological specific hemagglutination inhibition and electron microscopical methods. This is the first report on the isolation of coronaviruses in tissue culture from the respiratory tract of calves in the FRG. PMID- 2556872 TI - [Functional characteristics of the adrenals in the ontogeny of male water voles Arvicola terrestris with various fur colorings]. AB - Studies have been made of the effect of colour mutations on the adrenal function in growing male voles. In postnatal life, the level of 11-oxycorticosteroids in the blood exhibited similar changes in brown (wild phenotype) and black (mutant) voles. The highest level was found in 1-2-month animals; a decrease took place to the 6th month which was followed by a secondary increase at the age of 8 months. Corticosteron production in vitro is the highest in brown males 5-month. In black voles, after 2 months the production remains unchanged, being rather high. In 5-, 6- and 8-month voles, corticosteron production in vitro increased, but not decreased after prior incubation. In mutant voles, the adrenals are more sensitive to stimulating effect of ACTH (5 units per 1 g of the adrenals) and inhibitory effect of estradiol (40 ng/100 mg) in vitro. PMID- 2556873 TI - [Postsynaptic potentials in the myocardium of snails in the genus Helix]. AB - Cardioregulating neurones in the right parietal and visceral ganglia of the snail evoke postsynaptic potentials of various duration, amplitude and polarity in the auricular and ventricular myocardium. Inhibitory neurones with a marked background activity (1-2 imp/s) evoke IPSPs with a duration of 150-200 msec and a latent period of 160-220 msec in the auricle, these potentials being blocked by tubocurarine. EPSPs of approximately the same duration may be recorded in the ventricle during stimulation of the commanding neurones of the pneumostome LPa3 and PPa/3, as well as unidentified neurones. Action potentials in some other identified cardiostimulating neurones (PPa7, V1, V6) induce slow and sustained depolarization in the myocardium. Functional specificity of elements within fast and slow regulatory systems is suggested: discrete IPSPs and EPSPs account mainly for coordination of the systolic contractions of the auricle and ventricle, whereas long-lasting PSPs affect the frequency and intensity of the whole heart. PMID- 2556874 TI - [The effect of L-DOPA on potentials of the dorsal surface of the spinal cord evoked by the stimulation of primary afferents in newborn rat pups]. AB - In experiments on 5-30-day rat puppies, studies have been made of the effect of L DOPA (100 mg/kg, intraperitoneally) on the activity of interneurones of the dorsal horn of the spinal cord as revealed from the parameters of potentials of the dorsal surface of the spinal cord. The specific pattern of reaction in 5-day animals is manifested in a succession of inhibitory inhibition and increase in the activity of neurones monosynaptically activated by low-threshold afferents. Both the amplitude and duration of polysynaptic components of the potentials of the dorsal surface are rather high. From the 7th day, deep and stable inhibition is observed which is accompanied by a decrease in the amplitude of all components of the potential of the dorsal surface. At later stages of ontogenesis, a decrease is observed in the inhibitory effect of L-DOPA on the activity of interneurones which monosynaptically contact with low- and especially high threshold afferents; in contrast to earlier stages, but similar to adult animals, evident inhibition was revealed in the activity of interneurones which have polysynaptic contacts with high-threshold afferents and afferents of flexor reflex. Thus, within the first weeks of postnatal life, basic qualitative changes are observed in the pattern of the reaction of interneurones of the dorsal horn to exogenic catecholamines. PMID- 2556875 TI - [Basic patterns in the morphofunctional evolution of the monoaminergic structures of the hypothalamus]. PMID- 2556876 TI - [The evaluation and choice of treatment methods (surgical removal, proton-beam irradiation) in endosellar hypophyseal adenomas]. AB - The article analyses the immediate and late-term results of treatment of endosellar pituitary adenomas: removal through a transsphenoidal approach--16 patients, transnasal stereotaxic administration of 90Y--14 patients, irradiation by proton beam--263 patients. Surgery as well as radiotherapy produced quite satisfactory results. Proton therapy on a Leningrad synchrocyclotron is not inferior to the modern surgical methods in efficacy and safety. It is indicated in endosellar pituitary adenomas when the sella turcica measures no more than 20 x 16 mm. PMID- 2556877 TI - [The role of radiation therapy in treating hypophyseal adenomas]. AB - The article reviews various methods for the exposure of pituitary adenomas to the effect of radiation applied either as the only measure or in combination with surgery and drug therapy. The indications for radiotherapy are discussed according to the size and hormonal activity of the tumor. PMID- 2556878 TI - [Adenohypophysis function in Itsenko-Cushing disease before and after proton-beam irradiation]. PMID- 2556880 TI - Polyglycolic acid (Dexon) mesh for packing to control intra-abdominal bleeding. Case report. AB - Profuse bleeding from parasacral veins during abdominoperineal excision of the rectum for carcinoma was controlled by packing the pelvic cavity with polyglycolic acid (Dexon) mesh. The mesh pack could be left in place without giving rise to infection or otherwise interfering with the postoperative course. PMID- 2556879 TI - [Endocrine function in the late period after radiation therapy of acromegaly]. AB - The results of treatment of pituitary adenomas manifested by the acromegaly syndrome with catamnestic follow-up of up to 10 years are discussed. The dynamics of changes in acromegaly and various endocrine functions with the use of distance gamma therapy and proton beam irradiation were studied in the comparative aspect, on the basis of which the indications for these methods of radiotherapy in acromegaly were specified. PMID- 2556881 TI - Induction of glutathione S-transferase, placental type in T9 glioma cells by dibutyryladenosine 3',5'-cyclic monophosphate and modification of its expression by naturally occurring isothiocyanates. AB - The effect of dibutyryladenosine 3',5'-cyclic monophosphate (dibutyryl cAMP) on the expression of glutathione S-transferase placental type (GST-P) was examined in rat glioma cell line using an immunohistochemical technique. Cultured T9 glioma cells were negative for GST-P activity under normal conditions. However, treatment with 1 mM dibutyryl cAMP produced GST-P expression in about 50% of the cells, as well as some morphological changes. The expression of GST-P was increased with addition of dibutyryl cAMP together with 1 microgram/ml allyl isothiocyanate (AITC) or 0.1 microgram/ml benzyl isothiocyanate (BITC). With these combinated treatments, almost all cultured cells showed a strong positive reaction for GST-P, although AITC or BITC alone elicited GST-P in only 5% of the cultured cells. The results of the present study indicate that dibutyryl cAMP causes functional as well as morphological differentiation of T9 glioma cells. PMID- 2556882 TI - Focal myelin thickenings in a peripheral neuropathy associated with IgM monoclonal gammopathy. AB - Nerve biopsy in peripheral neuropathies associated with an IgM monoclonal gammopathy may occasionally display focal myelin thickenings. In a patient with such an IgM neuropathy, in whom an anti-myelin-associated glycoprotein (MAG) antibody activity was present in the serum, single-fiber preparations revealed 34% of internodes bearing myelin swellings. The morphometric, morphological and ultrastructural findings were reminiscent but not identical to those of the hereditary tomaculous neuropathy with liability to pressure palsies. Atypical features for tomacula included lack of spiralization of the redundant loops of myelin around the axons and their predominant external situation with regard to the myelin sheath. The frequent colocalization of myelin thickenings and the widening of myelin lamellae typical of IgM neuropathies, are highly suggestive of some pathogenetic link between the two abnormalities. The redundant loops of myelin in IgM neuropathies possibly result from a defect in the axon-myelin adhesion secondary to the binding of IgM on an epitope of MAG directly involved in cell-cell adhesion. PMID- 2556883 TI - Response to RIT 4237 oral rotavirus vaccine in human milk, adapted-and soy formula fed infants. AB - During the first month of life 28 full-term newborns were breast-fed (18 males and 11 females). Thereafter 8 infants continued breast-feeding while the remainder were randomly fed on either an adapted milk formula (n=13) or a soy formula (n=7). At five months, after an oral dose of RIT 4237 rotavirus vaccine of bovine origin was given, growth and IgM/IgG type antibodies against rotavirus were measured. Weight gain was similar in all infants. There were 2 IgM and 1 IgG responders out of 7 soy fed infants, compared with 4 out of 8 human milk fed (both IgM and IgG) and 7 out of 13 IgM and 6/12 IgG formula fed infants responding to vaccination. This observation confirms previous results obtained with polio, diphtheria tetanus and pertussis vaccines indicating that soy-protein formulas may interfere with immunization processes. PMID- 2556884 TI - Comparative study of neuroectodermal tumor and Ewing's sarcoma of the bone. Histopathologic, immunohistochemical and ultrastructural features. AB - We carried out a histologic, immunohistochemical and ultrastructural study of 19 tumors, previously classified as Ewing's sarcoma of the bone (ESB). The tumors were categorized histologically into four groups according to the presence or absence and the distribution of Homer-Wright rosettes, together with the immunohistochemical and ultrastructural features: 1) neuroectodermal tumor of bone (NTB) with diffusely dispersed rosettes (4 cases), 2) conventional ESB totally lacking the rosettes (9 cases), 3) intermediate-type tumor with characteristics of both NTB and ESB (4 cases), and 4) spindle cell-type tumor with focal rosettes (2 cases). Ultrastructurally, NTB contained neurotubules and neurosecretory granules, whereas ESB consisted of only undifferentiated cells. Immunoreactivity with antineural antibodies, such as Leu-7, and those against synaptophysin, NSE, neurofilament, and chromogranin, was detected in the majority of NTB, in intermediate- and spindle cell-type tumors and in two of the nine cases of ESB. In view of these findings, we propose that use of the term "neuroectodermal tumor of the bone" should be restricted to tumors with an extensive distribution of rosettes, and that those without unequivocal rosettes should be classified as ESB, although ESB may be the most undifferentiated form of NTB and some overlap may occur between the two tumor types. PMID- 2556885 TI - Secretory carcinoma coexistent with mucinous carcinoma in the breast. Report of a case. AB - Secretory carcinoma and mucinous carcinoma were found to coexist in the breast of a 67-year-old post-menopausal woman, although the tumors were separated by a thin fibrous septum. Histochemically, intra- and extracellular secretory materials in both carcinomas were strongly positive for alcian blue, PAS and mucicarmine staining, but immunohistochemically negative for alpha-lactalbumin and CEA. Membrane-bound intracytoplasmic vacuoles showing emiocytosis were observed in both the secretory and mucinous carcinomas by electron microscopy. No differences were observed between the tumor cells of secretory carcinoma and those of mucinous carcinoma by histochemistry, immunohistochemistry and electron microscopy. However, there were definite statistically significant differences in the results of morphometry of tumor cell nuclei. Secretory carcinoma is considered to be an anaplastic variant type of mucinous carcinoma. PMID- 2556886 TI - [Effect of propranolol on ouabain-sensitive Na+,K+-ATPase activity of the rat heat]. AB - Effect of propranolol on activity of basal ATP-ase measured in presence of Mg2+ activator, on that of total ATP-ase determined in presence of Mg2+, Na+, K+ activators and on that of Na+,K+-activated ATP-ase calculated from the difference of the previous two ones has been studied. Propranolol has not considerably influenced basal ATP-ase activity in 10(-7)-10(-4) M concentrations, 30-50% inhibition has been found in 1 X 10(-3)-5 X 10(-3) M concentrations. Effect of propranolol on total ATP-ase activity has been found to be dependent on doses and inhibition has been greater than inhibition experienced in case of basal ATP-ase. Enzyme kinetic investigations have shown that in case of basal ATP-ase propranolol has only decreased affinity to substrate but has not influenced rate of enzyme reaction, in case of total ATP-ase the affinity to the substrate has not changed but the reaction rate has significantly decreased. Ineffective concentrations of propranolol have potentiated inhibition effect of Na+,K+-ATP ase. The authors have supposed that the observed changes have been results of non specific membrane effects of propranolol and composition of membrane lipids may also play an important role. PMID- 2556887 TI - The role of endogenous and exogenous prostacyclin in the gastric mucosa under physiological and pathological circumstances. AB - The intracellular effect of exogenously administered prostacyclin in the gastric mucosa seems to be a polyphasic effect, namely: 1. Effect on the cyclic nucleotide (cAMP, cGMP), turnover; 2. Effect on the calmodulin-content; 3. DNA and RNA changes; 4. Influence on protein synthesis; 5. New cell formation. While the endogenous prostacyclin exerts a natural protection against damaging noxae. PMID- 2556888 TI - Selective decrease of duodenal phosphoprotein phosphatase activity is a general property of duodenal ulcerogens. AB - A phosphoprotein phosphatase (PPPase) is inhibited in rat duodenal mucosal cells very early after a single s. c. injection of the duodenal ulcerogens cysteamine, propionitrile and mepirizole. The effect seems to be independent of their effects on gastric acid secretion and on duodenal suppression of alkaline secretion. The enzyme inhibition is dose- and time-dependent under in vivo conditions. The inhibition of the PPPase activity is enzyme-selective at the level of mucosal cells in the duodenum. Under in vitro conditions, none of the duodenal ulcerogens inhibited PPPase activity. The results indicate that the effect of the ulcerogens on PPPase activity is probably exerted indirectly. When given simultaneously in vivo, propionitrile attenuated the inhibitory effects of cysteamine on the PPPase activity. However, both propionitrile and cysteamine potentiated the effect of mepirizole on PPPase depletion. These data indicate that cysteamine and propionitrile may act through the same mechanism when depleting PPPase activity. The mechanism of the decrease of duodenal protein phosphatase activity by mepirizole is probably different from the other duodenal ulcerogens. PMID- 2556889 TI - Free radicals and their interpretations. AB - Oxygen free radicals can be blamed for evoking gastric mucosal damage, because of the protective effect of some lipid soluble free radical scavengers (vitamin A related compounds, Vitamin E). Direct determination of free oxygen radicals related chemical entities in the gastric tissue during ulcerogenesis yielded controversial results. Aluminum antacid compound together with acid binding property exhibited cytoprotection too, elevating the tissue PGE2 level substantially. Magnesium containing antacid according to our model experiments on red blood cells damage by free radicals, is capable to bind free radicals as well as to counteract with the dangerous intracellular calcium accumulation. It has been concluded that aluminum-magnesium antacid has a cytoprotective effect via: 1. acid binding; 2. prostaglandin generation; 3. free radical scavenging; 4. calcium antagonist activity. PMID- 2556890 TI - Effect of ionic milieu to the free radicals generated by phenizine methosulphate (PMS). AB - The effect of free radicals generated by PMS was studied for membrane damage in the presence of different ions in the erythrocyte model. The degree of membrane damage depended on the quality of ionic composition in the incubation medium. We supposed that the degree of membrane damage depends on the average life and concentration and/or reactivity of the free radicals generated. For control of this supposition free radicals were generated by PMS in the presence of Sodium-di thionite in isosmotic, waterly systems with different ionic composition. At different time intervals the concentration of free radicals was measured by the ESR method. It seams that concentration of radicals depends on the qualitative composition of ionic milieu. The increase of the average life of free radicals generated by PMS is accompanied by decrease in their reactivity. This is reflected by a moderate membrane damage. PMID- 2556891 TI - Neuromuscular actions of endothelin on smooth, cardiac and skeletal muscle from guinea-pig, rat and rabbit. AB - The peptide endothelin (human, porcine) was investigated for effects on basal muscle tone and on responses to transmural nerve stimulation in a series of smooth muscle preparations, as well as in guinea-pig atrium and rat and guinea pig diaphragm. Endothelin lacked effect on basal tone or on spontaneous and electrically driven contractions in skeletal and atrial muscle. It contracted guinea-pig ileum, pulmonary and femoral arteries, rat anococcygeus, vas deferens and urinary bladder and rabbit taenia coli, whereas guinea-pig taenia was relaxed. Guinea-pig urinary bladder and vas deferens and rabbit iris sphincter were unaffected up to 3 x 10(-8) M. Endothelin thus has a unique pattern of smooth muscle effects, exhibiting mostly contractile but also relaxing effects. Endothelin modified contractile responses to transmural nerve stimulation, yielding marked and persistent enhancement, in guinea-pig and rat vas deferens, and enhancement also in guinea-pig pulmonary artery. In guinea-pig and rat vas deferens the response to exogenous ATP was increased by endothelin, thus suggesting a strong post-junctional enhancement of neurotransmission. In guinea pig ileum nerve-induced responses were inhibited by endothelin, whereas exogeneous acetylcholine was enhanced, an effect suggesting a simultaneous pre junctional inhibition and post-junctional enhancement. The Ca2+ channel blocker felodipine counteracted the stimulatory effects of endothelin on tone and transmurally induced contractions. Tachyphylaxis to endothelin action was sometimes evident, but the anococcygeus being less prone to this might be useful for studies on endothelin antagonism. Endothelin thus has prominent post junctional, and also probably pre-junctional, effects, lending further support for a distinct biological role of this peptide. PMID- 2556892 TI - Omega-conotoxin GVIA and nifedipine inhibit Ca2+ action potentials in rat locus coeruleus neurons. PMID- 2556893 TI - [Human papilloma virus in the male]. AB - Infection by human papillomavirus (HPV) is destined to constitute one of the most common reasons for consulting the urologist. The HPV play a fundamental role in the genesis of the uterine neck carcinoma. Recently entirely subclinical lesions of the male genitals have been demonstrated, which may only be diagnosed by means of optical magnification and application of acetic acid, as in lesions of the uterine cervix. We present the results of the study by means of this technique of 37 sexual partner of an equal number of women suffering from HPV infection, our diagnostic procedure and the location of the lesions found. The male displayed HPV lesions in more than 75% of the cases studied. Circumcision appears to reduce the incidence of affectation. No patient had symptomatology nor gave evidence of venereal disease at first sight. The sexual transmission of HPV is demonstrated as well as its responsibility in the genesis of various carcinomas, primarily female. Both members of the sexual couple should be studied and treated if we want to tackle this pathology guarantees of success. PMID- 2556894 TI - [Mucinous adenocarcinoma of the urachus]. AB - Malignant tumours of the urachus are very rare and they give certain difficulty in their differential diagnosis with vesical cupula adenocarcinoma. We present a case of mucinous adenocarcinoma of the urachus in a 75-year-old patient and we analyse the behaviour, clinical background, diagnosis and treatment of these tumours. PMID- 2556895 TI - [Urinary lithiasis composed of silica]. AB - We present a case of urinary silica (silicon dioxide) lithiasis in a patient without a background of medication use justifying the formation of this calculus. We review the literature on this type of lithiasis and comment on some aspects of the metabolism of silica connected with urolithiasis. PMID- 2556896 TI - Occult breast carcinoma in patients undergoing reduction mammaplasty. AB - Seven patients who had breast reduction surgery and whose preoperative physical examinations were unremarkable were found to have breast carcinoma. In the five in whom mastectomy was performed, most closures were difficult, and in one patient bilateral mastectomy was complicated by wound dehiscence. In only one of these seven was it possible to obtain information regarding the hormonal binding status of the tumor cells. These and other sequelae would not have occurred had the tumors been diagnosed before operation. Because physical examination alone is not sufficiently sensitive for the diagnosis of breast cancer, we suggest that mammography be included in the evaluation of patients consulting surgeons for breast reduction. PMID- 2556897 TI - Acceleration of rat brain beta-adrenoceptor subsensitivity following the coadministration of histamine receptor antagonists with imipramine. AB - Systemic administration of isoprenaline to rats produced a dose-dependent increase in water drinking which was effectively blocked by propranolol. This dipsogenic effect was significantly inhibited by the subacute (4 days) administration of imipramine (18.1 mg/kg/day) together with either the H1 histamine receptor antagonist, chlorpheniramine (0.1 or 1.32 mg/kg/day), or the H2-histamine antagonist, cimetidine (1.91 mg/kg/day) or ranitidine (0.60 or 1.51 mg/kg/day). The oral subacute administration of imipramine alone had no significant effect on this behavior. However, chronic ingestion of imipramine alone (21 days) caused a significant reduction in the isoprenaline-induced behavior. It is concluded that the desensitization of central beta-adrenoceptors, as evidenced by inhibition of isoprenaline-induced drinking, can be accelerated following the oral subacute co-administration of imipramine with either H1- or H2 histamine receptor antagonists. It is also seems the central histamine receptors may partially contribute towards the mechanism of antidepressant effect of imipramine. PMID- 2556898 TI - The release of interleukin-1-like activity by macrophages in two models of immunological inflammation in the rat. AB - Interleukin-1 (IL-1) is a putative mediator of inflammation released by activated macrophages in vitro. The IL-1 release by rat macrophages collected either from exudates in pertussis-induced air pouches or from the peritoneum during adjuvant arthritis has been investigated. In air pouch inflammation LPS-stimulated macrophages collected from sensitized animals release more IL-1 than cells from control rats at day 6 after challenge. This enhanced IL-1 release parallels the extent of mononuclear cell migration in the inflammatory lesion. In adjuvant arthritis LPS-stimulated macrophages collected from sensitized animals release more IL-1 than cells from control rats at days 16 and 23 after adjuvant injection. The secondary inflammation in arthritic rats was statistically significant at days 16 to 28. These results indicate that during immunological inflammation macrophages either from the inflamed area or from a non-inflamed region release more IL-1 than control cells. This release parallels the extent of the inflammation and may be important in its pathogenesis. PMID- 2556899 TI - Inhibitory effects of methyl 7-butyl-4,5,6,7-tetrahydro-3-methylamino-4,6-dioxo-5 propyl- 2H-pyrazolo[3,4-d]pyrimidine-2-carboxylate (AA-2379) on lysosomal enzyme and arachidonic acid release from rat polymorphonuclear leukocytes and its mode of action. AB - AA-2379 (methyl 7-butyl-4,5,6,7-tetrahydro-3-methylamino-4,6-dioxo-5-propyl- 2H pyrazolo [3,4-d]pyrimidine-2-carboxylate) has antiinflammatory, analgesic, and antipyretic activities, and inhibits the type III allergic (Arthus) reaction. In the studies reported here, we investigated the effect of AA-2379 on rat polymorphonuclear leukocyte (PMN) functions to clarify the mechanism of the antiinflammatory and antiallergic actions of AA-2379. AA-2379 at 10(-4) M inhibited lysozomal enzyme release. AA-2379 inhibits formyl methionyl-leucyl phenylalanine (fMLP)- and C5a-induced arachidonic acid release; their 50% inhibitory concentrations were 2.8 x 10(-5) and 3.8 x 10(-5) M, respectively. Because dibutyryl cAMP, a cAMP analogue, and 3-isobutyl-1-methylxanthine, a cAMP phosphodiesterase inhibitor, inhibited fMLP-induced arachidonic acid release, and AA-2379 inhibited cAMP phosphodiesterase and increased cAMP content in PMNs, it is likely that AA-2379 inhibited arachidonic acid release by increasing cAMP content in rat PMNs. Furthermore, from the studies of fMLP-induced arachidonic acid release in Ca free medium it is suggested that AA-2379 inhibits the process which depends on Ca concentration in the medium. These results suggest that the inhibitory effect of AA-2379 on inflammation and allergic reactions such as the Arthus reaction is partly exerted by inhibiting PMN functions such as arachidonic acid and lysozomal enzyme release. PMID- 2556900 TI - In vitro effects of leukotriene B4 (LTB4) on canine PMN effector function(s). AB - The canine has become an accepted research model for the examination of a number of human clinical conditions. Despite it's status as a research model, little is known regarding the peripheral effects of inflammatory mediator substances. Products of arachidonic acid metabolism (leukotrienes) are reported capable of altering leukocyte functions. Because of the emerging importance of the canine research model and leukotrienes we examined the effects of leukotriene B4 (LTB4) on several in vitro functions of isolated canine peripheral polymorphonuclear leukocytes (PMN). Changes in forward angle light scatter properties of the cells were used as one measure of PMN activation. Other functional changes examined following LTB4 pretreatment included chemotactic capability, the electrophysiological state of the cell plasma membrane, and the metabolic oxidative response (i.e. H2O2 production). Random cellular movement of PMNs increased by 120% and 72% following preincubation with 10(-7) and 10(-9) M LTB4, respectively. LTB4 between 10(-7) and 10(-13) M did not significantly alter cellular resting membrane potential. Between 10(-7) and 10(-9) M LTB4 elicited significant levels of cellular H2O2 production. Although significant, H2O2 production was less than 40% that induced by phorbol myristate acetate (PMA). In numerous respects, canine in vitro PMN responses parallel previous reports of human cell function(s) in the presence of inflammatory mediators and may represent an attractive alternative for investigation of PMN dysfunctions. PMID- 2556901 TI - The effect of the anabolic steroid, stanozolol, on the production of procollagenase by human synovial and skin fibroblasts in vitro. AB - The ability of the anabolic steroid, stanozolol, to stimulate procollagenase production by human synovial and skin fibroblasts was examined in an in vitro assay system. Stanozolol is used therapeutically to treat a variety of connective tissue and vascular disorders and its clinical effects suggest that it can modulate connective tissue breakdown. The results showed that stanozolol was capable, in a dose dependent manner, of significantly stimulating procollagenase production by skin fibroblasts. However, in three synovial fibroblast lines no evidence was found of increased collagenase production following treatment with stanozolol; although the synovial fibroblasts secreted significantly increased amounts of procollagenase in response to IL-1. These results may shed some light on the mechanism of action in vivo of stanozolol in the treatment of connective tissue disorders. PMID- 2556904 TI - Equine herpes virus 1 (EHV-1) in liver, spleen, and lung as demonstrated by immunohistology and electron microscopy. AB - Ten aborted foals, diagnosed as infected with Equine Herpes Virus 1 (EHV-1) on histopathological criteria, were examined for the presence of EHV-1 using immunohistology as the investigative instrument. The primary reagent was an antiserum specific for viral envelope glycoproteins. Immunohistology localised EHV-1 to areas of liver necrosis and to the cytoplasm of infected Kupffer cells and hepatocytes. Cytoplasmic immunolabelling was also prominent in reticular cells of the red pulp of the spleen and in intact and degenerated bronchiolar epithelium. Cytoplasmic immunolabelling was seen in morphologically unchanged cells and in cells containing intranuclear inclusion bodies. Three aborted foetuses with no histological signs of EHV-1 infection were negative when immunostained for EHV-1. Detection by electron microscopy of EHV-1 virions confirmed the EHV-1 specificity of the immunolabelling procedure. PMID- 2556902 TI - Eicosanoid production in rheumatoid synovitis. AB - Leukotriene B4 (LTB4) synthesis in rheumatoid synovitis was studied using peripheral and synovial fluid polymorphonuclear leukocytes (PMNs) and rheumatic synovial lining cells. No differences were found in LTB4 synthesis between peripheral PMNs from healthy volunteers and rheumatoid arthritis patients. When peripheral and synovial PMNs from the same RA patient were compared, arachidonic acid-induced LTB4 synthesis in synovial fluid PMNs was increased 1.7-7.2 fold, whereas the response to Ca ionophore A23187 stimulation was similar. This suggests 5-lipoxygenase stimulating factor(s) in inflamed joints. Rheumatic synovial lining cells in a primary cell culture produced small amounts of LTB4, the concentrations being less than 0.1 per cent of those of prostaglandin E2 (PGE2). PGE2 synthesis in synovial cells was increased when arachidonic acid or interleukin-1 was added to the culture, whereas LTB4 production remained unaltered. The present results suggest that in inflamed joints LTB4 originates mainly from PMNs whereas synovial lining cells are the source for PGE2. PMID- 2556903 TI - Inhibition of neutrophil response by curcumin. AB - Blood neutrophils, when exposed to appropriate stimuli, aggregate, degranulate and generate superoxide anion. Curcumin, a non-steroidal antiinflammatory agent, modulated these functions, depending upon the kind of stimulus used. It inhibited monkey neutrophil aggregation induced by chemotactic peptide fmlp and zymosan activated plasma (ZAP) but did not affect that induced by serum treated zymosan (STZ) and arachidonic acid (AA). Generation of O2- radical was inhibited by curcumin, when cells were stimulated by AA, STZ and fmlp. Curcumin inhibited the release of myeloperoxidase, an azurophilic granule marker enzyme. Release of lysozyme was less susceptible to inhibition by curcumin. The results suggest that curcumin interferes with neutrophil responses to various physiological stimuli and a part of its antiinflammatory action is mediated via inhibition of neutrophil function. Inhibition of neutrophil function by curcumin appears to be mediated via calcium dependent mechanisms. PMID- 2556905 TI - Demonstration of bovine viral diarrhoea virus antigens in cell cultures and in paraffin-embedded tissue sections by the peroxidase-antiperoxidase (PAP) technique using monoclonal antibodies. PMID- 2556906 TI - Effect of clot formation and retraction on spin-echo MR images of blood: an in vitro study. AB - Phantoms were constructed that contained red blood cell (RBC)-free clots in varying stages of clot retraction. MR images of these samples were compared with those of retracted whole venous blood clots and a fresh rat brain standard. Images were obtained at 0.3 T, 0.5 T, 1.0 T, 1.5 T, and 2.4 T with T1-, spin density- and T2-weighted spin-echo pulse sequences. The presence or absence of venous blood cells in the clot caused only minor differences in T2- and spin density-weighted images of the clots at or below 1.5 T. On T2-weighted scans, the retraction of the RBC-free clot resulted in a progressive decrease in signal intensity at 2.4 T. Fully retracted RBC-free clots were markedly hypointense relative to serum and ranged from slightly hyperintense to isointense with brain and venous clots at 0.5-1.5 T. There were no striking concomitant signal intensity changes on the spin-density- or T1-weighted scans, which could have caused the changes seen on the T2-weighted images of the clots. Our results indicate that the physical basis of these MR effects in the RBC-free clots is the concentration of plasma protein. The combined concentration of plasma protein and the tightly packed RBC proteins in the venous clots causes the strikingly similar MR appearance of venous and RBC-free clots on clinical images. These results do not demonstrate the presence of the previously postulated selective T2 relaxation of intracellular paramagnetic deoxyhemoglobin in these in vitro venous clots. PMID- 2556907 TI - MR imaging of the brain in patients with mucopolysaccharidosis. AB - MR imaging of the brain was performed in eight patients with mucopolysaccharidosis (MPS). Two had MPS I S, one had MPS IIA, two had MPS IIB, two had MPS IIIB, and one had MPS VI. In the patients with MPS IIA and MPS VI, T1 and T2 were prolonged in various areas of the cerebral white matter. These findings seemed to correspond with the development of pathologic changes in MPS, such as perivascular pits in the white matter observed on slices of the fixed brain. In the patients with MPS IIA and MPS IIIB, the white matter did not show the proper signal intensity, which suggested that myelination was insufficient and that infiltration or deposition of glycosaminoglycan had occurred; this was consistent with the association of these two types with mental retardation. In the patients with MPS I S, no intracranial abnormalities were detected on MR images. MR imaging of the brain may be used to obtain a differential diagnosis of the various types of MPS, to estimate the extent of mental retardation, and to monitor the progress of this disease. PMID- 2556908 TI - Three-dimensional gradient-recalled MR imaging as a screening tool for the diagnosis of cervical radiculopathy. AB - The purpose of the study was to implement and assess a fast-screening, three dimensional Fourier transformation (3DFT) MR sequence for the cervical spine. This sequence maintains adequate signal-to-noise and image contrast similar to gradient-recalled echo two-dimensional Fourier transformation (2DFT) imaging. Thirty patients with radiculopathy were examined with 3DFT gradient-recalled echo imaging. The data set consisted of 60 contiguous 1.5- to 2.0-mm-thick axial slices with a total coverage of 9 to 12 cm. In 10 patients, comparison was made with 4-mm-thick axial T1-weighted spin-echo 2DFT or gradient-recalled echo 2DFT studies. With the use of a volume acquisition, adequate signal-to-noise and image contrast similar to T2*-weighted gradient-recalled echo 2DFT acquisitions were obtainable. Coverage was improved despite the use of thinner sections without interslice gap. Thin-section 3DFT provided superior detail of acquired foraminal and spinal canal stenosis and disk morphology. Limitations included increased sensitivity to patient motion and "wraparound" artifact in the slice-select direction. Overall, diagnostic confidence was improved with 3DFT owing to the reduction of partial volume artifact. We have adopted this technique as the primary screening method for diagnosing cervical radiculopathy. PMID- 2556909 TI - All bran vs corn flakes: plasma glucose and insulin responses in young females. AB - Plasma glucose and insulin responses to breakfasts containing all bran or corn flakes were determined in 11 normal female volunteers aged 21-27 y. The test meals provided 50 g egg, 133 g orange juice, 129 g 2% milk, 180 g coffee, 5 g margarine, and 30 g available carbohydrate from either all bran or corn flakes with 19 g or 1 g of dietary fiber, respectively. Plasma glucose and insulin responses were higher for the corn flake breakfast and the incremental area under the glucose curve was 40% greater than that for the all bran breakfast (t test, p less than 0.05). These results are consistent with the reported glycemic indices for these cereals when consumed alone but the glucose responses differ to a lesser degree when the cereals are ingested with other foods. Thus, in this group of well-defined subjects the effect of all bran and corn flakes on plasma glucose responses is attenuated when these cereals are incorporated into mixed meals. PMID- 2556910 TI - Effect of breakfast cereals on short-term food intake. AB - We evaluated the effect of high-fiber cereals on short-term food intake. At 0730, 14 subjects ingested one of five cereals, plus milk and orange juice. At 1100 they were presented with a buffet lunch. There was a significant inverse correlation between fiber content of the cereals and energy intake at lunch. In a second study subjects ingested a very-high-fiber (VHF) cereal or a very-low-fiber (VLF) cereal. Fewer kcalories were ingested at lunch after ingestion of the VHF cereal than after ingestion of the VLF cereal. The degree of colonic microbial fermentation of the various cereals was evaluated by breath-hydrogen analysis. The higher-fiber cereals resulted in greater hydrogen production; however, this may not influence energy intake. The results of questionnaires that asked about hunger indicated that food intake can be reduced without the perception of feeling less hungry. Thus, we found that cereals containing relatively large quantities of dietary fiber may decrease short-term food intake. PMID- 2556911 TI - Dietary outcome in obese patients treated with a gastroplasty program. AB - Dietary intake was recorded on 7-d food registers by 18 patients for 2 y after horizontal gastroplasty (GP) for morbid obesity. The aim was to evaluate diet compliance and nutritional safety. In accordance with prescriptions, frequency of meals increased and amounts of food decreased. Contrary to intentions, qualitative improvements were minor and transient resulting in a lasting fractional increase of patients with inadequate intakes of a wide range of nutrients. Protein malnutrition could not be detected from measurements of serum albumin, plasma-prealbumin, or plasma retinol-binding globulin. Calcium was not included in the vitamin-mineral supplement and serum-Ca decreased. Despite thorough instruction, close follow-up, and gastrosurgery, there were no major qualitative dietary improvements. The study showed that bad compliance with an intended qualitative improvement of diet adds to the risks of being on a severely energy-restricted GP diet and increases the necessity for broad long-term supplements. PMID- 2556912 TI - A comparison of routine light microscopy, immunohistochemistry, and in situ hybridization for the detection of cytomegalovirus in gastrointestinal biopsies. AB - Eighty (80) paraffin-embedded intestinal biopsies from 35 AIDS patients were evaluated for the presence of cytomegalovirus by use of biotinylated DNA probe, immunohistochemical assay, and routine hematoxylin and eosin (H&E) staining. Cytomegalovirus was detected in 13 biopsies (16.3%) from five patients (14.2%). The distribution of positive biopsies was one in seven esophageal biopsies, three in 19 small bowel biopsies, six in 27 colonic biopsies, and three in 17 rectal biopsies. Immunoelectron microscopy was employed to confirm the presence of CMA in four of the positive in situ hybridization cases. In situ hybridization for CMV DNA was more sensitive than immunostaining, and both proved superior to routine H&E staining in terms of sensitivity. PMID- 2556913 TI - Biloma secondary to hepatocellular carcinoma in an HIV-seropositive patient. AB - Biloma has been defined as an extraductular collection of bile within a defined capsular space. Prior reports have documented an association of biloma with abdominal trauma and abdominal surgery. Biloma has not been reported in association with or as the presenting manifestation of hepatocellular carcinoma. In addition, hepatocellular carcinoma has not been previously reported in an HIV seropositive patient. We present the case of an HIV-seropositive patient with hepatocellular carcinoma complicated by a biloma. PMID- 2556915 TI - Cigarette smoke potentiates the DNA-damaging effect of manmade mineral fibers. AB - Epidemiological and experimental studies suggest that manmade mineral fibers (MMMFs) have DNA-damaging and carcinogenic properties. To investigate the hypothesis that cigarette smoke can potentiate MMMF-induced DNA damage, we exposed isolated calf thymus DNA to cigarette smoke condensate and/or three different types of MMMFs: rockwool, glasswool, and ceramic fibers. As an index of DNA damage, the hydroxyl radical-generated formation of 8-hydroxydeoxyguanosine (8OHdG) from deoxyguanosine (dG) was used. All the three fiber types, as well as cigarette smoke condensate alone, caused hydroxylation of dG residues in DNA, and, when smoke was combined with each of the different fibers, rockwool caused a synergistically increased formation of 8OHdG. We suggest that 1) iron-containing MMMFs such as rockwool are able to enhance synergistically cigarette smoke induced DNA-damage and 2) this damage is caused by hydroxyl radicals. PMID- 2556914 TI - Lymphatic and hematopoietic tissue cancer in a chemical manufacturing environment. AB - Nested case-control studies of non-Hodgkin's lymphoma (52 cases), multiple myeloma (20 cases), nonlymphocytic leukemia (39 cases), and lymphocytic leukemia (18 cases) were conducted within a cohort of employed men from two chemical manufacturing facilities and a research and development center. Exposure odds ratios were examined in relation to 111 work areas, 21 specific chemicals, and 52 chemical activity groups. Associations were observed for a maintenance and construction subgroup (non-Hodgkin's lymphoma) and a chlorohydrin production unit (nonlymphocytic leukemia). The odds ratio for the association of "foremen and others" with non-Hodgkin's lymphoma was 3.2 (CI95 = 1.47-7.2) based on 11 cases. A duration-response trend was observed for the chlorohydrin unit with three of four cases assigned 5+ years to that unit. An association between non-Hodgkin's lymphoma and assignment to strong acid alcohol production units (OR = 8.3; CI95 = 2.3-30.7) was not supported by a duration-response trend. Two highly correlated chemical groups, antioxidants (five cases) and nitriles (four cases), were over represented among multiple myeloma cases. A duration effect was observed. However, examination of work histories did not reveal common jobs or departments among these cases. PMID- 2556916 TI - Osteogenesis in bone defects in rats: the effects of hydroxyapatite and demineralized bone matrix. AB - Hydroxyapatite (HA) and Demineralized Bone Matrix (DBM) are being investigated as potential osteogenic agents with hopes that these substances can be used to induce bone formation in non-union fractures. This study was done to determine the relative effects of HA and DBM implanted as moldable phospholipid composites in bone defects that result in non-unions. We studied 22 ten-month-old Long-Evans male rats with 5.0 mm unilateral radial defects implanted with HA, DBM, and a combination of both substances. Control defects were left unfilled. Eight weeks after implantation, the histological sections demonstrated a decrease in bone formation with HA relative to controls. The HA crystals were encapsulated by connective tissue stroma made up of collagenous elements, fibroblasts, and blood vessels. There were no indications of bone formation within the fibrous stroma. 45Calcium, alkaline phosphatase, and bone gla protein (BGP) assays demonstrated a 16% increase in bone formation in rats implanted with DBM, an 80% decrease in groups implanted with HA (p = 0.01) and an 80% decrease with DBM plus HA (p = 0.01). Radiologic analysis corresponds well with histological and biochemical results. We conclude that osteogenesis in non-union defects is enhanced by the implantation of DBM, while HA interferes with bone formation in the rat model. In the presence of both substances, HA appears to impede new bone growth, negating any positive effects seen with DBM. PMID- 2556917 TI - Neuroendocrine and stress hormone changes during mirthful laughter. AB - Positive emotional activities have been suggested as modifiers of neuroendocrine hormones involved in the classical stress response. To detect changes in these components during a mirthful laughter experience, the authors studied 10 healthy male subjects. Five experimental subjects viewed a 60 minute humor video and five control subjects did not. Serial blood samples were measured for corticotropin (ACTH), cortisol, beta-endorphin, 3,4-dihydrophenylacetic acid (dopac)--the major serum neuronal catabolite of dopamine, epinephrine, norepinephrine, growth hormone, and prolactin. Repeated measures analysis of variance showed that cortisol and dopac in the experimental group decreased more rapidly from baseline than the control group (p = 0.011, p = 0.025, respectively). Epinephrine levels in the experimental group were significantly lower than the control at all time points (p = 0.017). Growth hormone levels in the experimental group significantly increased during baseline (p = 0.027) and then decreased with laughter intervention (p less than 0.0005), whereas, the controls did not change over time (p = 0.787). ACTH, beta-endorphin, prolactin, and norepinephrine levels did not significantly increase. The mirthful laughter experience appears to reduce serum levels of cortisol, dopac, epinephrine, and growth hormone. These biochemical changes have implications for the reversal of the neuroendocrine and classical stress hormone response. PMID- 2556918 TI - Mandibulofacial dysostosis or bilateral hemifacial microsomia with hearing loss, telecanthus, tetramelic postaxial hexadactyly, congenital hypotonia and lymphedema with joint hypermobility, and pigmentary dysplasia: a new syndrome? PMID- 2556919 TI - Pectoralis major defect and Poland sequence in second cousins: extension of the Poland sequence spectrum. AB - We report on a patient with congenital absence of the left pectoralis major muscle, whose second cousin had the full Poland sequence. This suggests that isolated pectoralis major muscle defect should be included in the spectrum of anomalies characterized as the Poland sequence, postulated to result from disruption of blood supply in the embryonic subclavian and vertebral arteries, the site and degree of obstruction determining the sites and severity of the resulting anomalies. Very few cases are familial; in these the family pattern is compatible with an autosomal dominant mutant gene with reduced penetrance or delayed mutation. PMID- 2556920 TI - Poland anomaly in mother and daughter. AB - We report on Poland anomaly in a mother and her daughter. Further family history was negative for abnormalities of the hands or the pectoralis major muscle. A review of published cases of familial Poland anomaly is presented. Implications concerning the possible etiology of familial cases of Poland anomaly are given. PMID- 2556921 TI - Recessive inheritance of a high number of sodium pump sites. AB - The number of sodium pump sites on erythrocytes was measured on 1,847 individuals in 80 Utah kindreds ascertained through probands with cardiovascular disease. Likelihood analysis supported recessive inheritance of high pump number. The major locus explained 14.0% of the variance in pump number; polygenic inheritance explained another 63.4%. Homozygotes for the recessive allele occurred with a frequency of 1.74% and had a mean pump number estimated as 566.0 sites/red blood cell (RBC) versus a mean of 312.2 sites/RBC for the other genotypes. Young individuals with the high pump number genotype were leaner, and older adults with the high pump number genotype were heavier. Diabetes and early hypertension were more prevalent in women with the high pump number genotype. Although not significant, obesity in adults of both sexes and early coronary heart disease in men were more prevalent in individuals with the high pump number genotype. PMID- 2556922 TI - Diagnostic dilemma of an unsuspected hyperfunctioning accessory parathyroid gland after total parathyroidectomy with autotransplantation in a peritoneal dialysis patient. AB - Persistent or recurrent hyperparathyroidism in patients with chronic renal failure may be a frustrating problem. We report a case history of a peritoneal dialysis patient who underwent total parathyroidectomy with autotransplantation for secondary hyperparathyroidism, developed tertiary hyperparathyroidism, and in an attempt to control hypercalcemia underwent seven partial resections of the autotransplant. Subsequently, a total excision of the parathyroid autograft was performed, but the patient continued to have hyperparathyroidism and unexpectedly was found to have a hyperplastic fifth parathyroid gland identified by thallium technetium subtraction scan. The fifth gland was removed, and a part was implanted in the right forearm; however, the autoimplant had to be completely removed because of rapidly developing hypercalcemia. Hypercalcemia was controlled, but elevated levels of parathormone persisted. Remaining parathyroid tissue could not be found. PMID- 2556923 TI - Severe visual loss related to isolated peripapillary retinal and optic nerve head cytomegalovirus infection. AB - We examined ten patients from a consecutive series of 73 patients with either isolated cytomegalovirus papillitis or limited cytomegalovirus retinitis contiguous with the optic disk. Patients with peripheral retinitis and other areas of retinitis were excluded. All patients were treated with ganciclovir. Two distinct types of cytomegalovirus infection of the peripapillary area were identified. Type I was characterized by spread of limited retinitis to the optic disk margin, good central visual acuity, and permanent arcuate and altitudinal visual field defects that enlarged and became more complete as the retinitis progressed toward the disk. Type II appeared to be a true cytomegalovirus infection of the optic nerve characterized by primary, isolated papillitis with peripapillary retinitis, an early afferent pupillary defect, and good initial visual acuity, which rapidly deteriorated despite prompt antiviral therapy. Peripapillary cytomegalovirus retinitis appears to be an important and underreported cause of visual morbidity in patients with AIDS. PMID- 2556925 TI - Detection of Epstein-Barr virus by in situ hybridization. Progress toward development of a nonisotopic diagnostic test. AB - This work presents some initial quantitation of an in situ hybridization method for detection of Epstein-Barr (EB) virus nucleic acids. The purpose is to develop evaluative criteria for diagnosis of viral presence in clinical tissue specimens. In this work simultaneous denaturation of probe and target DNA and an alkaline phosphatase conjugate to detect biotinated probe were used as described by Unger et al. For evaluation of the hybridization, a variety of cell lines, both productively and latently infected, that were hybridized in situ using nick translated 32P-labeled viral probe sequences and counted by scintillation after the method of Lawrence and Singer were used. Producer cells (B95-8) showed intense foci of staining in approximately 5% of cells, with most of the other cells showing varying staining intensity. Raji cells showed varying amounts of signal from cell to cell. Namalwa cells exhibited one spot in most cells that was decreased after cells were treated with Actinomycin D (dactinomycin, Merck Sharp & Dohme, West Point, PA). Signal was identified in only a third of these same cells after sectioning. EB virus-negative Ramos cells showed no signal. The nuclear punctate nature of the signal generated is diagnostic of infected cells, and may be a useful test for cultured cells or pathologic specimens. PMID- 2556924 TI - Development of osteoarthritic lesions in mice by "metabolic" and "mechanical" alterations in the knee joints. AB - Male, 10-week-old C57B1 10 mice received a single intraarticular injection in the knee joints with papain, iodoacetate, or collagenase. This led to osteoarthritic lesions, such as matrix depletion, chondrocyte proliferation, and osteophyte formation, in the injected knee joints within several weeks. After injection of iodoacetate and papain, the main osteoarthritic alterations were localized in the femoropatellar joint, whereas injection of collagenase led to marked osteoarthritic lesions in the femorotibial joint. The mechanism of induction of these alterations appears to differ for iodoacetate and papain on one site and collagenase on the other site. Data are presented that collagenase injection, by way of damaging ligaments and tendons, destabilizes the knee joint eventually leading to osteoarthritic alterations. In contrast, injection of papain or iodoacetate directly interferes with cartilage metabolism resulting in osteoarthritic changes. PMID- 2556927 TI - Human papillomavirus DNA in respiratory papillomatosis detected by in situ hybridization and the polymerase chain reaction. AB - The authors have demonstrated the presence of human papillomavirus (HPV) types 6 and 11 in 10 of 13 (77%) juvenile laryngeal papillomatosis by in situ DNA hybridization using as probes the radiolabeled DNAs of HPVs 6, 11, 16, and 18. Of six specimens from adult laryngeal papillomatosis assayed by the same technique, only 33% were positive. Immunohistochemistry to detect HPV capsid antigens performed on serial sections gave positive signals in 44% (8 of 18) of the specimens, all from juvenile lesions. These results were in agreement with in situ hybridization, except in two cases. When both series (juvenile and adult) were analysed by amplification of a 450-bp fragment corresponding to the L1 ORF of the HPV genomes directed by the polymerase chain reaction, the frequency of positive specimens rose to 100%. Our data agree with the concept that HPV is implicated in the etiology of laryngeal papillomatosis. PMID- 2556926 TI - Prognostic significance of CD30 (Ki-1/Ber-H2) expression in primary cutaneous large-cell lymphomas of T-cell origin. A clinicopathologic and immunohistochemical study in 20 patients. AB - The histologic and immunophenotypical features of 20 primary cutaneous large-cell lymphomas of T-cell origin were investigated and correlated with clinical data to obtain prognostically relevant criteria. Histologic evaluation, using the updated Kiel classification, showed that these large-cell lymphomas represent a morphologic spectrum, often making classification rather arbitrary. It is therefore concluded that the clinical relevance of histologic subtyping is limited for this group of lymphomas. Immunophenotypical studies revealed significant differences between CD30-positive and CD30-negative lymphomas. CD30 positive lymphomas generally presented with localized skin disease, and had a favorable prognosis (9 of 10 patients alive and in complete remission; median survival, 37 months). In contrast, CD30-negative lymphomas often presented with or rapidly developed generalized disease; all patients died of lymphoma (median survival, 17 months). These findings suggest that CD30 expression is an important prognostic parameter for this group of primary cutaneous large-cell lymphomas. PMID- 2556928 TI - Origin and relationship between different cell types in malignant fibrous histiocytoma. AB - The derivation of histiocyte-like cells in malignant fibrous histiocytoma (MFH) has been a matter of debate. To shed light on this problem two cell lines from two subsequent recurrencies of MFH were established. The existence of two different cell populations, mainly fibroblast-like in the first cell line and mainly histiocyte-like in the second, was shown by light and electron microscopy, DNA measurements, and karyotype analysis. By detailed banding analysis and identification of several identical chromosomal marker types in the two cell lines, it was proven that they originally derived from the same single cell or single clone. Because the first cell line, with mainly fibroblast-like cells, was in the hypotriploid region and the second, with mainly histiocyte-like cells, was in the penta-hexaploid region, the data explained the appearance of histiocyte like cells in MFH as a consequence of chromosomal progression. PMID- 2556929 TI - Antibody to type I insulinlike growth factor receptor inhibits growth of Wilms' tumor in culture and in athymic mice. AB - The role of the type I insulinlike growth factor (IGF) receptor in regulating growth of Wilms' tumor (WT) was evaluated by examining the effect of antibody mediated inhibition of this receptor on tumor growth in cell cultures and as heterotransplants in athymic mice. An antibody to the human type I IGF receptor (alpha IR-3) inhibited 125I-IGF-1 binding and prevented stimulation of thymidine incorporation by IGF-1 in vitro. Intraperitoneal administration of alpha IR-3 to nude mice bearing WT heterotransplants prevented tumor growth for 4 weeks and resulted in partial regression of established tumors. These data indicate the importance of IGF action in control of WT growth in vivo, and suggest potential therapeutic application using antigrowth factor receptor antibodies to block growth factor action. PMID- 2556932 TI - A role for the beta-subunit in the expression of functional Na+-K+-ATPase in Xenopus oocytes. AB - In all cellular systems studied so far, the catalytic alpha- and the glycosylated beta-subunit of Na+-K+-ATPase are coordinately synthesized and are assembled into stoichiometric alpha, beta-complexes. In contrast to these data, in this study we show that the fully grown oocyte of Xenopus laevis synthesizes much less beta subunit than alpha-subunit. The alpha-subunit produced in excess over the beta subunit is membrane associated but highly trypsin sensitive and can be compared with the immature alpha-subunit population identified in epithelial cells immediately after synthesis (K. Geering, J. P. Kraehenbuhl, and B.C. Rossier, J. Cell Biol. 105: 2613-2619, 1987). The Xenopus oocyte thus turns out to be a unique system to study the functional role of the beta-subunit. Injection of beta subunit-specific mRNA transcribed in vitro from a beta-cDNA clone (derived from Xenopus kidney, A6 cells) into oocytes results in translation of a glycosylated beta-subunit. The synthesis of this exogenous beta-subunit increases significantly the proportion of trypsin-resistant oocyte alpha-subunits able to perform cation-dependent conformational changes. In addition, 25-65% more ouabian binding sites are expressed at the plasma membrane in beta-mRNA-injected oocytes. In contrast, newly synthesized alpha-subunit translated after injection of size fractionated mRNA enriched in alpha-mRNA remains trypsin sensitive as the oocyte alpha-subunit. These data suggest that association of the beta-subunit to the alpha-subunit provokes a structural rearrangement of the alpha-subunit that might be a first step toward the functional maturation of the Na+-K+-ATPase and its expression at the plasma membrane. PMID- 2556931 TI - Role of Mg in the activation of Na-H exchange in dog red cells. AB - Depleting dog red cells of Mg prevents activation of Na-H exchange by cell shrinkage but has no effect on activation by acidification of the cytosol. Replacing cell Mg restores the activation of Na-H exchange by cell shrinking. We conclude that Mg is required, not for the transport process per se, but for the transduction of the volume stimulus. PMID- 2556930 TI - Molecular genetic analysis of lymphoid tumors arising after organ transplantation. AB - A variety of gene analyses were performed on lymphoid tumors from transplant patients who received cyclosporine A for immunosuppression. Epstein-Barr virus DNA was detected in the tumors, and the structure of circular episomal virus DNA was used as a measure of cell clonality. This analysis was correlated with clonality determined by study of immunoglobulin gene rearrangement. Some of the tumors had DNA rearrangements near the c-myc gene. Analysis suggested the pathogenesis of the tumors and indicated four categories of lymphoproliferation, three neoplastic and one reactive. PMID- 2556934 TI - Characteristics of two types of calcium channels in rat pituitary gonadotrophs. AB - The properties of Ca2+ channels in cultured rat pituitary gonadotrophs were analyzed by the patch-clamp technique. The inward Ca2+ currents, recorded in the presence of 5.2 mM Ca2+ or Ba2+, included a fast, transient component with activation-inactivation kinetics and a delayed component with slower activation. The midpoint of the activation curve lay at -30 mV for the transient component and at -12 mV for the delayed component. At the midpoint, changes in potential of 9.5 and 13 mV induced an e-fold change in the activation of the transient and delayed components, respectively. The rate of inactivation of the first component was strongly voltage dependent. At -43 mV, a 7.4-mV change in potential induced an e-fold change in the fraction of Ca2+ channels available to conduct Ca2+ current. During long-lasting (100-200 ms) low-frequency depolarizing voltage clamp pulses, the size of the delayed component of the Ca2+ current remained constant. The differential effects of membrane potential on inactivation and the different time constants for activation of the two components of the Ca2+ conductance indicate the presence of two types of Ca2+ channels in the membrane of the gonadotroph: the rapidly inactivating current appears to be attributable to a T-type channel, and the noninactivating current corresponds to the L-type channel described in many other cell types. PMID- 2556933 TI - Suppressive action of cobalt on exocytosis and respiratory burst in neutrophils. AB - Activation of exocytosis and respiratory burst in rabbit neutrophils by the chemotactic peptide fMet-Leu-Phe is inhibited by Co2+. Inhibition is antagonized by extracellular Ca2+ and is dependent on the time of preincubation of cells with Co2+ before addition of activator. Co2+ inhibits the enhancement of 45Ca association that occurs during activation with fMet-Leu-Phe. Interference with Ca2+ translocation across the plasma membrane by Co2+ is probably not the cause of inhibition of neutrophil activation, because activation in the absence of extracellular Ca2+ is inhibited by Co2+. Activation of neutrophils by phorbol myristate acetate is inhibited at higher Co2+ concentrations than activation by fMet-Leu-Phe. Inhibition of the superoxide production by Co2+ occurs both in the presence or in the absence of cytochalasin B. Fluorescence of neutrophils loaded with quin2 is diminished by Co2+, indicating that Co2+ had entered into the cytoplasm. The results are compatible with the view that Co2+ inhibits exocytosis and respiratory burst in neutrophils by an interaction with a Ca2+-dependent intracellular target. PMID- 2556936 TI - Role of calcium in mediating action of carbachol in T84 cells. AB - To examine the role of calcium in mediating carbachol's action in secretory epithelia, we simultaneously measured intracellular free [Ca] [( Ca]i) and transepithelial chloride transport in T84 cells grown on collagen-coated filters. [Ca]i was measured with fura-2 and fluorescence microscopy and expressed as a relative value [( Ca]'i) normalized to control. Chloride transport was measured as the short-circuit current (Isc) with a voltage clamp. Monolayers were pretreated with cyclic AMP to augment the response of Isc to carbachol, a procedure that did not qualitatively change the response of the monolayer to carbachol. The carbachol-induced changes in Isc appeared to be dependent on the increase in [Ca]i. First, carbachol caused both Isc and [Ca]'i to increase in parallel. Isc increased from 32 +/- 5 to 70 +/- 9 microA and then declined to 57 +/- 16 microA while [Ca]'i increased from 72 +/- 14 to 156 +/- 22 nM and then declined to 133 +/- 45 nM. Second, the carbachol-induced increases in Isc and [Ca]'i were correlated. The greater the hormone-stimulated rise in [Ca]'i, the higher the increase in Isc. Third, carbachol's stimulation of Isc was blunted by preventing the calcium spike with the cellular calcium buffer 1,2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetra-acetic acid (BAPTA). Although the carbachol induced increase in [Ca]'i appeared necessary for the increase in Isc, it was not clear if carbachol's action was solely the result of an increase in [Ca]'i. Increasing [Ca]'i with ionomycin, although causing Isc and [Ca]'i to increase in parallel, failed to increase Isc to the levels observed with carbachol. These experiments suggest that although the carbachol-induced increase in Isc is dependent on the increase in [Ca]i, the hormone may activate a second process that increases the sensitivity of the calcium-activated transport process to changes in [Ca]i. PMID- 2556935 TI - Role of endothelium in endotoxin blockade of voltage-sensitive Ca2+ channels in smooth muscle cells. AB - Coculture of endothelial and smooth muscle cells was used to study effects of endotoxin on depolarization-induced Ca2+ transients in fura-2-loaded individual smooth muscle cells. Although endotoxin did not modify the response of cultured smooth muscle cells to depolarization, endotoxin resulted in an attenuation of cytosolic Ca2+ (Cai2+) transients in response to K+ depolarization and failure of KCl-induced contractions in smooth muscle cells when they were cocultured with endothelial cells. The observed endothelial modulation of smooth muscle responses was not accomplished via gap junctions. The possible role of free radical species secreted by endothelial cells in conditioning of smooth muscle responses to depolarization was supported by the results of three sets of experiments: 1) endothelial cells did respond to endotoxin with oxidative burst, 2) pretreatment of cocultured cells with catalase prevented endotoxin-induced downregulation of Ca2+ transients, and 3) in isolated smooth muscle cells, the addition of hydrogen peroxide virtually abolished depolarization-induced Ca2+ transients. Hence vascular endothelium stimulated by endotoxin generates reduced oxygen intermediates, which in turn downregulate depolarization-induced Cai2+ transients in smooth muscle cells. This phenomenon may contribute to the development of hypotension in septicemia. PMID- 2556937 TI - Atrial natriuretic peptide stimulates submandibular gland synthesis and secretion of cGMP. AB - Binding of atrial natriuretic peptide (ANP) to rat submandibular gland and its effect on guanosine 3',5'-cyclic monophosphate (cGMP) formation and salivary secretion were investigated. Membranes rapidly and specifically bound 125I-ANP. Binding was inhibited by unlabeled ANP (IC50 approximately 1.6 nM), but not by atriopeptin I, other COOH- and NH2-terminal deleted ANP fragments, or agents such as pilocarpine or substance P. Scatchard analysis revealed a single class of high affinity sites (dissociation constant 0.74 +/- 0.25 nM; maximal binding capacity 20.5 +/- 6.3 pmol/mg protein). Intravenous infusion of ANP with pilocarpine caused a significant dose-dependent increase in the levels of cGMP detected in plasma and saliva. Because salivary cGMP may have originated in plasma, the effect of ANP on cGMP formation was evaluated in dispersed cells. ANP evoked a concentration-dependent increase in both cGMP synthesis and secretion (EC50 approximately 1.7 x 10(-8) M). The atrial peptide did affect basal or l isoproterenol-stimulated adenosine 3',5'-cyclic monophosphate synthesis in dispersed cells. When infused by itself and/or with pilocarpine, ANP did not alter the rate of spontaneous or pilocarpine-induced salivary flow, secretion of chloride, or protein release. The data demonstrate the presence of guanylate cyclase-coupled ANP receptors in submandibular gland; the atrial peptide, however, does not exert an effect of the secretory function of the gland. PMID- 2556938 TI - Effects of fiber type and training on beta-adrenoceptor density in human skeletal muscle. AB - The density and distribution of beta-adrenergic receptors in type I and II fibers of human gastrocnemius and quadriceps muscles were characterized in ten healthy sedentary subjects and in a subgroup of six subjects before and after 12 wk of endurance exercise training. Total tissue content of beta-receptors was measured in frozen sections of skeletal muscle biopsies incubated with 125I-labeled cyanopindolol in the presence and absence of 10(-5) M L-propranolol. The relative beta-receptor densities of type I and II fibers were delineated autoradiographically. Muscle fiber types were identified in adjacent serial sections by histochemical staining of myofibrillar adenosine-triphosphatase (ATPase) activity. Type I fibers had a threefold greater beta-receptor density than type II fibers of the same muscle [P less than 0.001; type I-to-type II fiber ratio of beta-receptor density was 3.06 +/- 0.43 (SD)]. Exercise training elicited a change in muscle fiber subtype composition (+34% type IIa and -42% type IIb; P less than 0.05 and P = 0.066, respectively), a 40% increase in citrate synthase activity of skeletal muscle (P = 0.01), and a 23% rise in peak oxygen uptake (P less than 0.001). However, no change in total tissue content of beta-receptors was observed after exercise training, even when receptor density was adjusted for preconditioning fiber type composition. Thus beta-receptor density of type I fibers of human skeletal muscle is threefold greater than that of type II fibers. Enhanced capacity for aerobic metabolism after endurance exercise training is not associated with upregulation of total beta-receptor density. PMID- 2556939 TI - Development in rats of the brain-pituitary-adrenal response to hypoglycemia in vivo and in vitro. AB - To clarify the nature of the stress hyporesponsive period that occurs in neonatal rats, the development of the response of the brain-pituitary-adrenal axis to hypoglycemia stress in rats was assessed in vivo and in vitro. Hypothalami were removed from the brains of neonatal (9-35 days postnatal) or adult rats and incubated in vitro for sequential 30-min periods in Krebs buffer for determination of corticotropin-releasing factor (CRF) secretion under conditions of altered glucose concentrations. As expected from previous studies, CRF secretion from adult hypothalami was significantly increased in severely hypoglycemic conditions (0.55 mM glucose) by approximately 50% above base-line values (in 5.5 mM glucose). However, lowering glucose did not elicit an increase in CRF release from hypothalami of rats less than 35 days of age. Hypothalami obtained from rats less than or equal to 24 days old also failed to show consistent secretory responses to potassium depolarization. At 35 days postnatal the response to hypoglycemia was significant and similar to the adult response. To determine if the lack of hypothalamic response to hypoglycemia in vitro could be correlated with the in vivo responses to hypoglycemia, rats aged 4 days to adult were injected intraperitoneally with porcine insulin and killed at different times after injection. Insulin injections lowered plasma glucose levels in fasted 4-day-old rats in a dose-dependent fashion, but a nadir in glucose (approximately 40 mg/dl) was not reached until 90 min; the same treatment produced a nadir in glucose within 30 min in fasted rats 10 days old and older, suggesting that the 4-day-old rats are relatively insulin insensitive.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556940 TI - Intracellular pH regulation in IEC-6 cells, a cryptlike intestinal cell line. AB - Regulation of intracellular pH, pHi, was studied using microspectrofluorimetry of the pH-sensitive, fluorescent dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein in the rat intestinal crypt cell line, IEC-6. In N-2-hydroxyethylpiperazine-N'-2 ethanesulfonic acid (HEPES)-buffered solutions with pHi 7.25, treatment with a pulse of NH4Cl caused cells to acidify and then recover to control level. Because recovery was Na dependent, blocked by 1 mM amiloride, and unaffected by the presence and absence of Cl, it was likely because of a Na+-H+ exchanger. Cells were also acid loaded by changing from HEPES to HCO3-CO2-buffered solutions. pHi again recovered, but 1 mM amiloride reduced the rate of H+ efflux by only 47%. This HCO3-dependent, amiloride-insensitive H efflux required Na+ but not Cl- and was completely blocked by 200 microM [H2] 4,4'-diisothiocyanostilbene-2,2' disulfonic acid (DIDS). We conclude that a Na+-HCO3- cotransporter was operative. Cl-free solutions caused pHi to increase from 7.19 to 7.41; this effect required the presence of exogenous HCO3-CO2 but not Na and was blocked by 200 microM [H2]DIDS. A Cl- -HCO3- exchanger is the most likely explanation for these data. All the pHi regulatory mechanisms are operative in NaCl-HCO3-CO2-buffered solutions. The Na+-H+ and Na+-HCO3- mechanisms are acid extruders, whereas the Cl -HCO3- exchanger is an acid loader. These transporters may be important for generating HCO3 secretion by intestinal crypt cells. PMID- 2556941 TI - Intracellular pH dependence of buffer capacity and anion exchange in the parietal cell. AB - When parietal cells (PC) are stimulated with histamine, the anion exchanger rate increases three to five times to compensate for alkaline loading induced by H+-K+ adenosinetriphosphatase (ATPase) and to provide Cl for acid secretion. It has been hypothesized that this increased activity is caused by the increase in intracellular pH (pHi) that often occurs in stimulated PC (from 7.1 to a maximum of 7.3). The dependence of the anion exchanger on pHi was studied using microspectrofluorimetry of the pH-sensitive dye 2',7'-bis(carboxyethyl)-5(6) carboxyfluorescein (BCECF). N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES)-buffered solutions were used because the anion exchanger can transport OH (or HCO3) in exchange for Cl- even with [HCO3]o = 200 microM. It was found that when solutions were changed either from NaCl to Cl- free or Cl- free to NaCl, rates of change of pHi (delta pH/delta t) were strongly dependent on pHi: nearly 0 at pHi 6.6 and 1.25 pH/min at pHi 8.0. To convert these pHi changes into anion flux rates, the intrinsic buffer capacity (beta i) was determined over the same pHi range by making small changes of [NH4]o to determine the resulting changes of [NH4]i and pHi (i.e., beta i = delta[NH4]i/delta pHi) in PC that had been pretreated with 1 mM amiloride and 200 microM [H2]4,4'-diisothiocyanostilbene 2,2'-disulfonic acid (DIDS) [to block Na+-H+ and Cl- -OH-(HCO3-) exchange]. beta i was also strongly dependent on pHi: at pHi 6.5 beta i = 48 mM/pH, and this decreased as pHi increased; at pHi 7.75 beta i = 8 mM/pH. The derived anion fluxes (i.e., JOH = beta i x delta pH/delta t) were roughly linearly related to pHi between 6.6 (JOH near 0) and 8.1 (JOH = 13 mM/min). Between pHi 7.1 and 7.3, the range normally observed during stimulation of PC, rates of anion exchange increased by 75%. This pHi sensitivity cannot explain the 300-500% increase in anion exchanger activity observed during secretagogue stimulation of PC. PMID- 2556942 TI - Cobalamin release from intrinsic factor and transfer to transcobalamin II within the rat enterocyte. AB - To ascertain the mechanism of release of cobalamin (Cbl) from intrinsic factor (IF) and subsequent formation of transcobalamin II (TC-II)-Cbl complex, we studied the intracellular distribution of 57Co-labeled Cbl after its uptake in suckling and adult rats. The amount of Cbl bound to IF, to the IF-Cbl receptor via IF, and to TC-II was determined by immunoprecipitation with monospecific antisera raised to these proteins. IF-Cbl receptor activity was found to be very low in suckling rats up to 12 days after birth. Oral administration of leupeptin in amounts known to alter protein turnover had no effect on the release of Cbl from IF nor did it inhibit the formation of the TC-II-Cbl complex in either adult or suckling animals. However, oral administration of chloroquine resulted in a transient increase in the intestinal concentration of Cbl in both adult and suckling rats and in total inhibition of Cbl released from IF in adults rats. Chloroquine prevented completely the transfer of Cbl to TC-II in adult rats and inhibited the transfer by 50% in suckling rats. These data demonstrate that in adult mucosa utilizing receptor-mediated endocytosis, Cbl is transferred from IF to TC-II. This transfer does not require the IF-Cbl receptor, as it occurs in suckling rats. Finally, transfer of Cbl to TC-II is decreased by a drug that alters vesicular pH. Because Cbl can be released at acid pH from IF, it is proposed that release of Cbl from IF and its transfer to TC-II occurs in an acidic vesicle. PMID- 2556943 TI - Relationship of lymph node metastasis to primary tumor size and microscopic appearance of roentgenographically occult lung cancer. AB - We investigated the relationship of lymph node metastasis to primary tumor size and microscopic appearance in 92 resected specimens obtained from patients with roentgenographically occult lung cancer (ROLC) located at a site along the airway between the main bronchus and the sub-subsegmental bronchi. Most of the patients were discovered by mass screening. All were treated surgically after bronchoscopic localization of cancer. The bronchial tree of the resected specimens was serial-sectioned into 2-mm thick blocks from the margin of resection to the sub-subsegmental bronchi. Bronchial wall invasion was noted in some blocks of all the specimens. The length of longitudinal extension (LLE) was defined as the product of the thickness and the number of consecutive blocks involved, counting from the most proximal to the most distal block. LLE was used as primary tumor size. Hilar and mediastinal lymph nodes were examined in 84 patients who underwent lymph node dissection. No nodal involvement was found in 59 cancers with LLE of less than 20 mm. Of 25 cancers with LLE of 20 mm or more, six showed nodal involvement. Eleven in situ carcinomas and four cancers of the "suspicious for invasion" type showed no lymph node metastasis. We contend that no lymph node dissection is required when pulmonary resection is performed for patients with ROLC if it is in situ carcinoma, if it is of the "suspicious for invasion" type, or if the LLE is smaller than 20 mm. PMID- 2556944 TI - Multicentric independent development of hepatocellular carcinoma revealed by analysis of hepatitis B virus integration pattern. AB - Two separate nodular lesions of hepatocellular carcinoma (HCC) were surgically removed from a patient with positive serum hepatitis B surface antigen. Southern blot analysis of DNA from the two separate HCC lesions showed different restriction patterns of integrated hepatitis B virus DNA, indicating their independent origins. The two lesions were composed mainly of poorly differentiated HCC; however, well-differentiated HCC was also observed in the periphery of the lesions. This histological finding is consistent with the multistep and independent development of HCC in each site of the liver. PMID- 2556946 TI - Hyperosmolality due to antacid treatment. AB - Magnesium trisilicate mixture is an antacid used commonly in our Intensive Care Unit in the prevention and treatment of stress ulcers. In this case the administration of large doses over a period of time led to the development of massive hyperosmolality, cerebral dehydration and coma. Management with hypotonic fluid resulted in complete recovery. PMID- 2556945 TI - Antinuclear antibodies in the sera of patients with nasopharyngeal carcinoma. AB - We studied the production of heterophile antinuclear antibodies (ANAs) in the sera of 50 patients, 20 with nasopharyngeal carcinoma (NPC) and 30 with other head and neck cancers (laryngeal cancer and maxillary cancer), before and after radiation therapy. A higher incidence of ANAs was found in the sera of patients with NPC and ANA production in these patients was higher after radiation therapy. We therefore performed in vitro experiments to explore the mechanisms of ANA production in the serum of postirradiated NPC patients. X-ray-irradiated NPC derived cells (NPC-KT) produced a large amount of Epstein-Barr virus (NPC EBV) compared with non-irradiated NPC-KT cells. Nasopharyngeal carcinoma EBV-infected lymphocytes produced high levels of ANAs. These data suggest that lymphocytes infected by EBV from NPC cells may produce ANAs in the sera of NPC patients. PMID- 2556948 TI - Interaction between enalapril and propofol. PMID- 2556947 TI - Isoflurane for thymectomy in myasthenia gravis. AB - The clinical and electromyographic effects of isoflurane were studied in eight myasthenic patients undergoing trans-sternal thymectomy. After inhalational induction of anaesthesia, intubating and operating conditions were good. Recovery from anaesthesia was rapid with minimal postoperative residual muscle weakness or respiratory depression. All patients were extubated within the first postoperative hour. Integrated electromyographic monitoring of the train-of-four response of adductor pollicis demonstrated that myasthenics are more sensitive than nonmyasthenics to the neuromuscular depressant effects of isoflurane. Recovery of the integrated electromyographic response was incomplete despite a satisfactory clinical recovery. PMID- 2556949 TI - [Hormonal and metabolic criteria of the adequacy of anesthesia during surgery of nephroblastoma in children]. AB - The data are presented on changes in hypophyseal-adrenal system and carbohydrate metabolism during transperitoneal nephrectomy in 42 children aged 1 to 15 years with stage II-III tumors. In 22 patients surgery was performed under combined neuroleptanalgesia with halothane for induction and maintenance of anesthesia. In 20 patients halothane was replaced by diazepam. Glucose, insulin, ACTH, CTH, hydrocortisone, lactic and pyruvic acid levels, total activity of LDG, its isoenzymes and cAMP have been assessed at different stages of anesthesia and surgery. PMID- 2556950 TI - [Results and experiences with several methods for the immunodiagnosis of amebiasis]. AB - Results and experiences of some immunodiagnostic methods for amebiasis. Experiences concerning the laboratory diagnosis of invasive amebiasis are described and discussed. Technics used include the Passive Hemagglutination Test with E. histolytica antigen (Behring FRG) and sheep erythrocytes as carrier and the Indirect Fluorescence Antibody Reaction (IFAR) with corpuscular antigen prepared from cultures of E. histolytica strain HK 9. Comparatively for a part infected hamster liver (Dr. Richle strain) has been used as antigen. We conclude that IFAR with HK 9 antigen is more sensitive than the other tests. It has been proved useful in the diagnosis of extraintestinal amebiasis (antibody titer greater than or equal to 1:160). PMID- 2556951 TI - Arterial infusion of combination therapy using dibutyryl adenosine 3',5' monophosphate and mitomycin C for hepatocellular carcinoma occluding main portal vein: case studies. AB - Intraarterial infusion therapy combining dibutyryl adenosine 3',5'-monophosphate and mitomycin C was administered to 8 patients with hepatocellular carcinoma occluding the main portal vein. Three patients survived more than nine months, including 1 who has been well for forty-eight months with technical imaging showing complete regression. A decreased level of serum alpha-fetoprotein of more than 90% was obtained in 38% of patients. Causes of death included hepatic failure, rupture of esophageal varices, acute gastric ulcer, and cerebral hemorrhage. It is concluded that the combination therapy may be useful for HCC. PMID- 2556952 TI - The effect of vitamin K supplementation on circulating osteocalcin (bone Gla protein) and urinary calcium excretion. AB - STUDY OBJECTIVE: To determine whether vitamin K administration affects urinary calcium excretion in postmenopausal women. DESIGN: Before- and after-trials with a 2-week treatment period. SUBJECTS: Healthy postmenopausal women (55 to 75 years old) were recruited from the convents in and around Maastricht. Controls (25 to 40 years old) were healthy premenopausal volunteers. INTERVENTION: Daily administration of 1 mg of vitamin K for 2 weeks. MEASUREMENTS: Serum immunoreactive osteocalcin: hydroxylapatite binding (HAB) capacity of serum immunoreactive osteocalcin; excretion of calcium, hydroxyproline, and creatinine in the urine during the last 2 h of a 16-h fasting period. RESULTS: In premenopausal women, no effect of vitamin K administration was seen. In the postmenopausal group, vitamin K induced increased serum immunoreactive osteocalcin concentration; normalization of the HAB capacity of serum immunoreactive osteocalcin (this marker was less than 50% that of the controls in the pretreatment samples); a decrease in urinary calcium excretion, notably in the "fast losers" of calcium; and a parallel decrease in urinary hydroxyproline excretion in the fast losers of calcium. CONCLUSIONS: The serum immunoreactive osteocalcin level may vary with vitamin K status. This variance should be taken into consideration if osteocalcin is used as a marker for osteoblast activity. Vitamin K is one factor that may play a role in the loss of bone mass in postmenopausal osteoporosis. PMID- 2556953 TI - [Curative radiotherapy of unresectable bronchial cancer. Apropos of 266 cases without metastasis with 4 to 6 years follow-up (oat cell carcinoma excluded)]. AB - Two hundred and sixty-six patients with inoperable bronchial carcinomas (oat cell excluded) without apparent metastases were treated with curative doses of radiotherapy. The malignancies were: 72% epidermoid epitheliomas, 11.5% adenocarcinomas, 10% large anaplastic cells and 6.5% without histology. According to the TNM classification, 3.5% were T1, 44% were T2 and 52% were T3. Mediastinal invasion was found in 44% of the cases. The mean age of the patients was 65 years (range 31-90 years). The Karnofsky index was less than or equal to 70 in 40% of the subjects. Unoperability was linked to the extent of local involvement in 59% of the cases and to age, general condition or an insufficient forced expiratory volume in 55% of the patients. Sixty to 65 Gy were administered, either classically fractionated or hypofractionated. After irradiation, 43% of the subjects underwent complete radiological remissions. Fifty-five percent of the patients died with local tumor evolution, 40% with isolated local evolution without associated metastases. The overall survival rate at 5 years was 7%; it was 16% for patients with a normal mediastinum and a Karnofsky index greater than 70 (51 cases); if these parameters were inversed, it was 2.5%. The fractionation protocol, the histological type, the use of chemotherapy (48 cases) did not affect the local control rate nor survival. There were almost no complications with this therapy. Patients frequently experienced functional improvement when curative doses of radiotherapy were applied to all the inoperable, non-metastatic cancers and the percentage of survivors at 5 years was non-negligible. PMID- 2556954 TI - [Focus on research and results of unusual radiotherapy technics (heavy particles, hyperthermia, radiosensitizers)]. AB - Heavy particles have been used in transcutaneous radiotherapy for almost 50 years. If these particles and pi mesons remain the domain of highly specialized centers, neutron therapy and, to a lesser degree, proton therapy can be considered common clinical protocols; their limits, complications, efficacities and costs are sufficiently well known to encourage several centers to acquire the necessary equipment. Their practical advantages involve a limited number of patients however, for the patients implicated, the prognoses are appreciably improved by the application of these therapies. Hyperthermia-associated radiotherapy is limited by the poor technical capabilities of the currently available hyperthermia machines. If biological experimentation was highly promising, clinical results to date have been unfulfilling. Nonetheless, hyperthermia-associated interstitial radiotherapy represents an interesting clinical possibility. Radiosensitizers are being extensively developed. The results of randomized therapeutic trials are still contradictory and the toxicity of these products renders their use difficult. New products are being tested; their lower toxicity and potent radiosensitization capacity are encouraging for the future use of this class of compounds. PMID- 2556955 TI - [Lymphoid tumor of the cavum in HIV infection]. PMID- 2556956 TI - [Oculomotor paralysis and peripheral neuropathy caused by IgM kappa monoclonal immunoglobulin. Transient effect of plasmapheresis]. PMID- 2556957 TI - [Sweat gland tumors in Niger. Anatomopathology]. AB - The authors report 15 cases of sweat gland tumour observed over 2 years in Niger where their pathology was never studied. These tumours accounted for 0.38 p. 100 of all examinations performed during the same period at the pathological laboratory of the University of Niamey. They ranked sixth among the diagnoses of skin pathology. Only routine histological techniques were used. The diagnoses obtained could be divided into 5 groups. Eight women presented with partly solid cystic tumours located on the legs in 5 cases; their structure was typical of nodular hidradenoma. Two male patients had cyst-pitted tumours the superficial portion of which showed a structure of papillary eccrine adenoma, while the deep portion contained a multitude of cysts with atrophic walls; the authors consider this type of tumour as a variant of papillary eccrine adenoma. One patient had a syringocystadenoma which also included a large cyst. Finally, one patient presented with a chondroid syringoma in the axillary region, and 3 patients had histologically aggressive carcinomas located on the sole, forehead and axillary region respectively. Clinical and paraclinical investigations failed to detect any deep malignancy in these three patients, and their tumours were regarded as compatible with sweat gland carcinomas. In this series, two lesions were unusually numerous: carcinomas and nodular hydradenomas. The first named probably are a consequence of the socio-economic conditions prevailing in the population of Niger: poverty prevents patients from consulting for diseases which produce no symptoms, which artificially increases the apparent incidence of aggressive lesions. As for nodular hydradenomas, they are large owing to the presence of cysts.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2556958 TI - [Thymus hormones in dermatology]. PMID- 2556959 TI - Genetic analysis of C4 polymorphism by use of DNA amplification (PCR), allele specific oligonucleotide probes and allele-specific restriction enzymes. AB - In vitro DNA amplification allows multiplication of selected gene segments thereby improving the sensitivity in DNA analysis. Different allelic variants in the amplified DNA may be disclosed either by subsequent hybridization with allele specific oligonucleotides or by subsequent allele-specific digestion with selected restriction endonucleases, followed by separation in agarose gel electrophoresis. The genes that code for human complement component C4 are polymorphic. Presently we demonstrate that allelic differences in C4, involving one base pair only, can be efficiently identified in the amplified DNA by each of the two techniques. A combination of both techniques may also be employed. The DNA amplification procedure may give access to selected 'haploid' fragments for individual DNA studies. PMID- 2556960 TI - Anti-herpesvirus activity of citrusinine-I, a new acridone alkaloid, and related compounds. AB - Citrusinine-I, a new acridone alkaloid isolated from the root bark of the citrus plant (Rutaceae), exhibited potent activity against herpes simplex virus (HSV) type 1 and type 2 at low concentrations relative to their cytotoxicity; 50% effective concentrations (ED50) of citrusinine-I were 0.56 micrograms/ml and 0.74 micrograms/ml against HSV-1 and HSV-2, respectively. Inhibitory action was also demonstrated against cytomegalovirus (CMV) and thymidine kinase-deficient or DNA polymerase mutants of HSV-2. The compound markedly suppressed HSV-2 and CMV DNA synthesis at concentrations which did not inhibit the synthesis of virus-induced early polypeptides. However, citrusinine-I had no inhibitory activity against HSV and CMV DNA polymerases in cell-free extracts. Although the target of this inhibitor remains to be elucidated, the most plausible candidate is a virus-coded ribonucleotide reductase. Citrusinine-1, when combined with acyclovir or ganciclovir, synergistically potentiated the antiherpetic activity of these agents. Based on a comparative study of the antiherpetic activity of citrusinine 1 and 28 related compounds, a structure-activity relationship could be established. PMID- 2556961 TI - Effects of diet composition on intake by adult wild European rabbits. AB - The voluntary dry matter intake (DMI) of several grass and legume diets, and the amount of dry matter (DM), nitrogen, fibre, and energy assimilated from each diet (i.e. the digestibility coefficients) are presented for the wild European rabbit Oryctolagus cuniculus. The DMI may be predicted from DM% and percentage of total nitrogen (on a DM basis) for a high DM diet (90-95%) but the general relationship may stand for fresh forage also. The metabolizable energy of a diet is correlated with DMI through the DM% and the percentage of nitrogen and fibre in the diet on a DM basis. The DM digestibility coefficient is correlated with fibre content. The wild rabbit's high efficiency of protein digestibility and low fibre digestibility compared with ruminants is also a characteristic of the domestic rabbit. It is suggested that the proportions of different nutrients required by wild rabbits are similar to those required by domestic animals. PMID- 2556963 TI - Electron spin resonance spectrum of Tyr-151 free radical formed in reactions of sperm whale metmyoglobin with ethyl hydroperoxide and potassium irridate. AB - A five-line ESR spectrum was observed at room temperature in reactions of sperm whale metmyoglobin with ethyl hydroperoxide (EtOOH) at pH 9.5 and with potassium irridate at pH's 7.0 and 9.5. A spectrum with the same g value and hyperfine splitting constant appeared in a reaction of sperm whale apomyoglobin with potassium irridate and was assigned to a tyrosyl radical on the basis of optical spectrum data obtained under the same reaction conditions. It was concluded that this radical arose from Tyr-151 for the following reasons. (i) This ESR spectrum could not be observed in the reaction of horse heart metmyoglobin, which lacks Tyr-151. (ii) Sperm whale metmyoglobin no longer gave this spectrum when treated with tetranitromethane (TNM) under conditions in which approximately one tyrosine is lost in sperm whale metmyoglobin but none is lost in horse heart metmyoglobin. (iii) A complex ESR spectrum observed in the reaction of sperm whale metmyoglobin with EtOOH at neutral pH was found to be a mixture of this five-line spectrum and one arising from an unidentified free radical formed in the reaction of horse heart metmyoglobin with EtOOH. The TNM-treated sperm whale metmyoglobin gave the same ESR spectrum as that observed in the reaction of horse heart metmyoglobin with EtOOH. PMID- 2556962 TI - Immobilization of pig muscle aldolase on a silica-based support. AB - Pig muscle aldolase was covalently attached to a silica-based support possessing aldehyde functional groups. The activity of the immobilized enzyme was 37 U/g solid, and the specific activity calculated on a bound protein basis was 1.9 U/mg protein. The optimum pH for the catalytic activity was pH 7.5. The apparent optimum temperature was found to be 45 degrees C. The Km app value of the immobilized aldolase with D-fructose 1,6-diphosphate as substrate was 1.25 X 10( 4) M. The conformational stability was improved by the immobilization. The immobilized aldolase was used for the continuous splitting of D-fructose 1,6 diphosphate. PMID- 2556964 TI - The chromophore retinal hinders passive proton/hydroxide ion translocation through bacteriorhodopsin. AB - Experiments have been performed to examine any influence of the chromophore retinal in bacteriorhodopsin (BR) on the passive proton/hydroxide ion flux through this integral membrane protein. BR was reconstituted into dimyristoylphosphatidylcholine (DMPC)-phosphatidylserine or DMPC dimyristoylphosphatidylglycerol unilamellar vesicles with molar lipid to protein ratios ranging from 30 to 150. The entrapped fluorescence dye pyranine served as a reliable indicator of the internal proton concentration. Transmembrane pH gradients were quickly established across the vesicular membrane and the kinetics of the induced fluorescence changes were compared for vesicles with incorporated native BR, BR bleached to the chromophore-free protein bacterioopsin, and BR regenerated from bacterioopsin with all-trans-retinal, respectively. For aggregated protein molecules, the H+/OH- diffusion across bacterioopsin was always considerably faster than that through the protein containing covalently bound retinal. The decay rate of the imposed pH-gradient was 4.4-9.1 and 2.0-5.1 times slower for native and regenerated BR, respectively, as compared to bacterioopsin. Stepwise regeneration of bacterioopsin with all-trans-retinal revealed a linear dependence of the predominant delta pH-decay time on the degree of regeneration. Essentially the same observations were made with monomeric protein molecules in vesicular lipid membranes. The results demonstrate that the chromophore retinal itself blocks the H+/OH- conducting pathway across the transmembrane protein BR or indirectly controls this path by inducing conformational changes in the protein upon binding. PMID- 2556966 TI - Structural transitions of carboxymethylated cytochrome c: calorimetric and circular dichroic studies. AB - Circular dichroism and differential scanning calorimetry studies on the unfolding refolding process of native and carboxymethylated cytochrome c, induced either by temperature or chemical agents, have been performed. The results have shown that the modified protein has a decreased conformational stability with respect to the native state, in agreement with a structure less compact, but still highly folded, which behaves as a thermodynamically stable "intermediate" between native and fully unfolded cytochrome c. PMID- 2556965 TI - Roles of Ca2+ in O2 evolution in higher plant photosystem II: effects of replacement of Ca2+ site by other cations. AB - The properties of the S2 states formed in Ca2+-extracted, Sr2+-substituted, and Cd2+-substituted photosystem II (PSII) were comparatively studied by means of thermoluminescence (TL) and low temperature EPR spectroscopy. The following results were obtained: (i) Ca-extracted PSII showed neither EPR multiline nor g = 4.1 signals but showed an abnormal TL band with an upshifted peak temperature, which did not oscillate on excitation with two or more flashes. Addition of Ca2+ to the extracted PSII reversed most of these effects concomitant with marked restoration of O2 evolution, but the EPR g = 4.1 signal remained lost. (ii) Sr2+ substitution largely restored the g = 4.1 signal and partly restored the multiline signal in a modified form having a reduced hyperfine line spacing concomitant with partial restoration of O2 evolution. Sr2+ substitution also reversed the abnormal TL peak temperature to normal, but the oscillation of the restored TL peak was much damped. (iii) Cd2+ substitution restored neither of the two EPR signals nor O2 evolution but reversed the abnormal TL peak temperature to normal. However, the reversed TL peak did not show any oscillatory behavior. Given these results, the function of the weakly bound Ca2+ in S-state turnovers in higher plant PSII is discussed. PMID- 2556967 TI - Metabolism of leukotriene E4 to 5-hydroxy-6-mercapto7,9-trans-11,14-cis eicosatetraenoic acid by microfloral cysteine-conjugate beta-lyase and rat cecum contents. AB - Leukotriene E4 was incubated with cysteine-conjugate beta-lyase isolated from the intestinal bacterium Eubacterium limosum. The reaction was terminated by addition of iodoacetic acid or dimethyl sulfate, and the products formed were isolated by reverse-phase high-performance liquid chromatography. The structures of two adducts of a metabolite were determined by uv spectroscopy, by gas-liquid radiochromatography, and by comparisons with chemically synthesized reference compounds. They were 5-hydroxy-6-S-carboxymethylthio-7,9-trans-11,14-cis eicosatetraeno ic acid (iodoacetic acid adduct) and 5-hydroxy-6-S-methylthio-7,9 trans-11,14-cis-eicosatetraenoic acid (dimethyl sulfate adduct) indicating that the structure of the underivatized metabolite was 5-hydroxy-6-mercapto-7,9,11,14 eicosatetraenoic acid (5,6-HMETE). The latter product is formed by beta-lyase catalyzed cleavage of the cysteine C-S bond in leukotriene E4. Leukotriene E4 was also metabolized to 5,6-HMETE by rat cecal contents. A product formed was trapped as the iodoacetic acid derivative and identified as 5-hydroxy-6-S-carboxy methylthio-7,9,11,14-eicosatetraenoic acid. It is concluded that intestinal leukotriene E4, originating from biliary excretion of systemic cysteinyl leukotrienes or produced in the intestine, is converted by microfloral cysteine conjugate beta-lyase to 5,6-HMETE. PMID- 2556968 TI - NADH-dependent microsomal interaction with ferric complexes and production of reactive oxygen intermediates. AB - The interaction of NADPH with ferric complexes to catalyze microsomal generation of reactive oxygen intermediates has been well studied. Experiments were carried out to characterize the ability of NADH to interact with various ferric chelates to promote microsomal lipid peroxidation and generation of .OH-like species. In the presence of NADH and iron, microsomes produced .OH as assessed by the oxidation of a variety of .OH scavenging agents. Rates of NADH-dependent .OH production were 50 to 80% those of the NADPH-catalyzed reaction. The oxidation of dimethyl sulfoxide or t-butyl alcohol was inhibited by catalase and competitive .OH scavengers but not by superoxide dismutase or carbon monoxide. NADH-dependent .OH production was effectively catalyzed by ferric-EDTA and ferric diethylenetriaminepentaacetic acid (DTPA), whereas ferric-ATP and ferric-citrate were poor catalysts. All these ferric chelates were reduced by microsomes in the presence of NADH (and NADPH). H2O2 was produced in the presence of NADH in a reaction stimulated by the addition of ferric-EDTA, consistent with the increase in .OH production. The latter appeared to be limited by the rate of H2O2 generation rather than the rate of reduction of the ferric chelate. NADH dependent lipid peroxidation was much lower than the NADPH-catalyzed reaction and showed an opposite response to catalysis by ferric complexes compared to .OH generation as production of thiobarbituric acid-reactive material was increased with ferric-ATP and -citrate, but not with ferric-EDTA or- DTPA, and was not affected by catalase, SOD, or .OH scavengers. These results indicate that NADH can support microsomal reduction of ferric chelates, with the subsequent production of .OH-like species and peroxidation of lipids. The pattern of response of the NADH-dependent reactions with respect to catalytic effectiveness of ferric chelates and sensitivity to radical scavengers is similar to that found with NADPH. Many of the metabolic actions of ethanol have been ascribed to production of NADH as a consequence of oxidation by alcohol dehydrogenase. Since the cytosol normally maintains a highly oxidized NAD+/NADH redox ratio, it is interesting to speculate that increased availability of NADH from the oxidation of ethanol may support microsomal reduction of iron complexes, with the subsequent generation of reactive oxygen intermediates. PMID- 2556969 TI - Resting state respiration of mitochondria: reappraisal of the role of passive ion fluxes. AB - Rat liver mitochondria respiring under resting state conditions in the presence of oligomycin were rapidly blocked with cyanide and the dissipation of the membrane potential, measured with a tetraphenylphosphonium-sensitive electrode, was followed over time. The plot of the rate of membrane potential dissipation versus the actual value of the membrane potential was nonlinear and identical to the plot of resting state respiration (titrated with small amounts of a respiratory inhibitor) versus the membrane potential. The relationship between the respiratory chain activity and the proton-motive force in mitochondria oxidizing succinate with either oxygen or ferricyanide as electron acceptors was also found to be identical. These results are interpreted as an indication that the passive permeability of the inner mitochondrial membrane toward ions is far more significant in maintaining resting state respiration than is the molecular slippage of the pumps in the respiratory chain. These results also confirm the non-ohmic characteristics of the inner mitochondrial membrane. PMID- 2556970 TI - Chemoembolization of liver malignancies--the experience at the university hospital in Frankfurt/M. (West-Germany). PMID- 2556971 TI - [Pilot combination phase II study of mitomycin C plus cisplatin for non-small cell lung cancer]. AB - Fifteen patients (six patients with adenocarcinoma, seven patients with squamous cell carcinoma, and two patients with large cell carcinoma) with advanced non small cell lung cancer (NSCLC) were evaluable for mitomycin C (MMC; 8 mg/m2 day 1, 8, every 3-4 weeks) plus cisplatin (CDDP; 80 mg/m2 day 1, every 3-4 weeks). Ten patients had had prior chemotherapy. Among 15 evaluable patients, no patient achieved complete response, and two patients showed partial response. The response rate of MMC plus CDDP against NSCLC was 13.3%. Toxic effects included anorexia (80%), nausea and vomiting (67%), leukopenia (53%), anemia (47%), nephrotoxicity (47%), thrombopenia (27%), liver injury (27%), and fever (7%). These toxic effects were reversible and manageable. The combination of MMC and CDDP appears to be valuable regimen against advanced NSCLC. PMID- 2556972 TI - [HCFU and 5-fluorouracil levels in the blood and tissue of hepatocellular carcinoma after oral administration of HCFU]. AB - HCFU was orally administered to 14 patients with hepatocellular-carcinoma, (including 11 patients with liver cirrhosis) and evaluated of HCFU and 5 fluorouracil (5-FU) levels. Blood and tissue 6-8 hr. after oral administration. The concentration of 5-FU in tissue was almost in the effective levels. In addition, the 5-FU level in the tissue of hepatocellular carcinoma tended to be higher than in non cancerous portion of the liver. 5-FU tissue concentration was not correlated with various laboratory data for the liver function (K-ICG, T. bil, GOT, GPT, etc.) From these results, it is suggested that HCFU is a useful anticancer agent for hepatocellular carcinoma especially for the cases accompanied liver cirrhosis. PMID- 2556973 TI - [Antitumor effects of cisplatin, cyclophosphamide and interferon-gamma (gamma IFN) against argyrophil small cell carcinoma of the uterine cervix heterotransplanted into nude mice]. AB - We investigated antitumor effect of cis-diammine-dichloroplatinum (cisplatin or CDDP), cyclophosphamide (CY) and interferon-gamma (gamma-IFN) against argyrophil small cell carcinoma (ASCC) of the uterine cervix using heterotransplanted ASCC tumor (YIK-1) into nude mice, containing HPV 16 DNA in a multicopy integrated form. No tumor growth retardation was observed in the nude mice which received the single administration of CDDP 2 mg/kg, CY 10 mg/kg or gamma-IFN 1 x 10(7) U/kg. However, the combined administration of CDDP and CY, or CDDP, CY and gamma IFN markedly inhibited the tumor growth. Moreover, with comparison about relative tumor volume between these two groups, the combination of CDDP, CY and gamma-IFN was more effective than that of CDDP and CY. These data suggest that the combination chemotherapy of CDDP, CY and/or gamma-IFN was effective for the suppression of tumor growth in argyrophil small cell carcinoma of the uterine cervix. Further study on the antitumor potentiation of various anti-cancer agent and the combination of them against ASCC is needed. PMID- 2556974 TI - [A resected case of hepatocellular carcinoma effectively treated by hepatic artery embolization and systemic chemotherapy]. AB - A 64-year-old male complaining of abdominal fullness was admitted to hospital for close examination. Hepatocellular carcinoma was diagnosed by various imaging techniques and the patient was treated by 3 transarterial embolizations and 2 courses of systemic chemotherapy with CDDP. The tumor was reduced and the effect was judged to be a partial response to these therapies. By resection, a residual mass had histologically no live cancer cells. This case was considered to have had a complete response with multidisciplinary treatment. PMID- 2556976 TI - New outlook on pericardial substitution after open heart operations. AB - The difficulties of reoperation owing to adhesions are well known. Clinical attempts to solve this problem using synthetic materials or glutaraldehyde-fixed pericardial xenograft have been less than satisfactory. Although experimental animal results have been good, they have not considered the influence of cardiopulmonary bypass (CPB) on adhesion formation. This study addressed the influence of CPB on the formation of adhesions and evaluated biodegradable polyglycolic acid as a material to reduce adhesions and as a pericardial substitute. Forty-five weanling sheep received implants of pericardial xenograft and polyglycolic acid with and without CPB. The pericardial xenograft showed no adhesions when implanted without CPB, but severe adhesions formed and a thick fibrinous layer covered the heart when CPB was used, making identification of coronary arteries at reoperation very difficult. The polyglycolic acid mesh implanted without CPB was absorbed and replaced with newly formed host collagen. With CPB, the polyglycolic acid was more rapidly absorbed, and a thinner layer of host collagen formed. Therefore, future animal studies must include CPB. In agreement with reported clinical results, glutaraldehyde-fixed pericardial tissue implanted with CPB resulted in severe epicardial reaction and therefore is not an ideal pericardial substitute. PMID- 2556975 TI - 15-hydroxyeicosatetraenoic acid (15-HETE) specifically inhibits the LTB4-induced skin response. AB - 15-Hydroxyeicosatetraenoic acid (15-HETE), a 15-lipoxygenase product of arachidonic acid, inhibits leukotriene B4 (LTB4)-induced chemotaxis of polymorphonuclear leukocytes (PMNs) in vitro. In this study the effects of intradermal injections of LTB4 were determined in the absence or presence of 15 HETE. For comparison intradermal injections of purified human complement split product C5a were performed in the absence or presence of 15-HETE. The skin response was evaluated by measuring the diameter of the wheal, the area of the flare and by intensity of the erythema (erythema index). LTB4 and C5a were injected at the concentration of 200 ng/ml. At this concentration the maximal skin response of LTB4 and C5a were equivalent. In contrast to C5a reaction, which resolved within 1 h, LTB4-induced skin response lasted up to 18 h. In all subjects the skin response was significantly decreased when LTB4 was injected together with 300 ng of 15-HETE. The decrease of wheal, flare, and erythema index averaged 81.9%, 56.6%, 53.6%, respectively, when all parameters were obtained at the maximal skin response. In contrast, the C5a-induced skin response was not affected by addition of 15-HETE, even when the final dose of 15-HETE was increased 10 times to 3 micrograms. The LTB4-induced reaction could last up to 18 h after injection. After the addition of 300 ng of 15-HETE the skin response resolved after 1 h. The present results demonstrate that 15-HETE is a specific inhibitor of the LTB4-induced skin response and brings additional evidence in support of the ability of 15-HETE to regulate the proinflammatory effects of LTB4 in vivo. PMID- 2556977 TI - Enalapril-induced cough. AB - A retrospective analysis of records from an outpatient medical practice was undertaken to determine the incidence and features of cough resulting from the use of enalapril maleate. Of 209 patients taking enalapril, 22 (10.5%) required discontinuation of therapy because of an intractable, dry cough. Cough was more than twice as common in women; 16 (14.6%) of 109 women and 6 (6%) of 100 men stopped taking enalapril because of cough. The cough resolved in 21 of 22 patients within 2 weeks of discontinuation of enalapril therapy. When the patients with cough were compared with the others, there was no significant difference in age, smoking status, creatinine levels, enalapril dosage, associated cardiopulmonary disease, or concomitant administration of medications. Among the 187 study patients who did not discontinue taking enalapril because of cough, many developed a persistent, dry cough that to date has not been severe enough to require discontinuation of therapy, after a mean follow-up period of 16 months. The enalapril-induced cough is insidious, dry, persistent, benign, and reversible on discontinuation of therapy. It is important to distinguish enalapril-induced cough from cough resulting from acute illness, reactive airway disease, and congestive heart failure. Optimal clinical application of enalapril in the treatment of hypertension and congestive heart failure will require increased awareness of this incessant cough, which requires discontinuation of the therapy in about 10% of patients. PMID- 2556978 TI - Polyvalent immune globulin and cytomegalovirus infection after renal transplantation. AB - To determine whether polyvalent, immune globulin (IgG) prevents cytomegalovirus (CMV) infection after cadaver renal transplantation, 28 patients were randomly allocated to receive 12 weekly infusions of 500 mg/kg of IgG (n = 15) or no treatment (n = 13). Both groups were similar with respect to age, sex, antigen mismatches, number of diabetics, and pretransplant donor/recipient CMV antibody titers. Moderately severe CMV infections occurred in 10 (77%) of 13 control subjects compared with 8 (53%) of 15 IgG-treated patients (not significant). Among those who developed CMV infections, prophylactic IgG had no effect on the severity or duration of fever, leukopenia, or hepatic enzyme elevations. Since none of the IgG-treated patients or control subjects in this study developed life threatening CMV complications, a beneficial effect of prophylactic IgG in the small number of patients at risk for more severe CMV infections could not be excluded. However, this investigation suggests that the routine, prophylactic administration of polyvalent IgG, to prevent or to ameliorate CMV infection in unselected cadaver renal transplant recipients, is not warranted. PMID- 2556979 TI - Latex and vinyl examination gloves. Quality control procedures and implications for health care workers. AB - In December 1987, we investigated an increased number of cases of herpetic whitlow in medical intensive care unit nurses who routinely gloved for secretion contact. One particular brand of vinyl examination glove had been used in the medical intensive care unit. Restriction endonuclease mapping established the similarity of employee isolates with one patient isolate of herpes simplex virus type I. When initial viral assay demonstrated 2.5% to 10% penetration of herpes simplex virus type I across unused gloves, an evaluation of glove quality was undertaken. In a 300-mL watertightness test, seven brands of vinyl gloves failed 4% to 28% (average, 11.1%; 132/1200), while seven brands of latex gloves failed 0% to 2.6% (average, 1.4%; 24/1750). The brand of vinyl glove that had been in use in the medical intensive care unit failed 28% of the time. Watertight gloves were then tested for permeability to herpes simplex virus type I. None of the latex gloves failed (n = 1726), while only 10 of the vinyl gloves failed (n = 1068, 0.95%). Extreme variability in glove quality was observed. However, gloves made from intact vinyl may provide similar protectiveness as those made from intact latex. As the demand for gloves increases, emphasis should be placed on the production of plentiful, better quality latex and vinyl gloves. PMID- 2556980 TI - [Immunohistochemical studies on pituitary adenomas in Wistar rats. 1. Demonstration of ACTH, LH, neurophysin, oxytocin and vasopressin in the pituitary of Ico:WIST rats from chronic toxicity studies]. AB - Spontaneous pituitary adenomas are common in certain strains of the laboratory rat. Investigations of Wistar rats of two years chronic toxicity studies revealed pituitary tumors in 50% of the females and 26% of the males. The morphology of the spontaneous changes in the pituitary gland was investigated with immunohistochemical and histological methods. The peroxidase-antiperoxidase (PAP) technique was used to localize different hormones (LH, ACTH) in cells of the pars intermedia and pars distalis as well as neurophysin, oxytocin and vasopressin the terminals of the classic neurosecretory system of the pars nervosa. The results show that most of the neoplasms were endocrinologically inactive chromophobe adenomas of the pars distalis. PMID- 2556981 TI - Destabilization of herpes simplex virus type 1 virions by local anesthetics, alkaline pH, and calcium depletion. AB - The infectivity of herpes simplex virus type 1 (HSV-1) was found to be markedly reduced by treating virions with the tertiary amine local anesthetics lidocaine, dibucaine and tetracaine. These treatments induced a characteristic shift in the buoyant density of the HSV-1 particles from a "light" to a "heavy" population. HSV-1 virions were unstable at alkaline pH, and alkali treatment caused the same shift in buoyant density. Ca2+ stabilized the "light" population. These results suggest that the physicochemical status of the HSV-1 envelope which is sensitive to the treatments described above plays an important role for the integrity of the virion. PMID- 2556983 TI - [Hypoglycemic polyneuropathy: report of a case with insulinoma]. AB - A case of a young man who presented symptoms and clinical signs of polyneuropathy that occurred in connection with recurrent hypoglycemic episodes is reported. The hypoglycemia was probably caused by a pancreatic islet tumor. There were symmetric weakness and wasting of hands and feet, absent tendon reflexes and 'glove and stocking' loss of sensation. Electromyographic studies showed denervation potentials with slight reduction of nerve conduction velocities. Sural nerve biopsy studied by optic and electronic microscopy showed axonal degeneration without signs of demyelination or remyelination. There are only 30 similar cases reported in the literature. According to experimental findings, the authors believe that glucopenia is the mechanism responsible for the development of the neuropathy, and that at present time there is no evidence for a direct insulin effect. PMID- 2556984 TI - The crystallinity of human deciduous teeth in hypophosphataemic vitamin D resistant rickets. AB - Five teeth were obtained from three patients with hypophosphataemic vitamin D resistant rickets (HVDRR) and five corresponding sound teeth from five healthy children. According to powder X-ray diffraction analysis, the half-peak breadths of (310) and (002) reflections of HVDRR dentine were smaller than those of normal dentine. Splitting fractions obtained from i.r. spectral analysis of HVDRR dentine powder were larger than those of normal. Microbeam X-ray diffraction analysis showed that the relative half-peak breadths of globular dentine in ground sections of HVDRR teeth were smaller than those of normal circumpulpal dentine. Transmission electron microscopy demonstrated that the hydroxyapatite crystals of globular dentine in HVDRR teeth were larger than those of normal dentine. Thus the crystallinity of deciduous tooth dentine in HVDRR was greater than that of normal dentine, mainly because of the large hydroxyapatite crystals in HVDRR globular dentine. PMID- 2556985 TI - Separation and partial characterization of three forms of collagenase inhibitor from bovine gingiva. AB - Three forms of collagenase inhibitor were isolated; one bound to Con A-Sepharose and the other two did not. The Con A-bound (Mr 38,000) and the two unbound (Mr 50,000 and 22,000) inhibitors contained about 20, 15 and 65% of the total inhibitory activity, respectively. The bound and one of the unbound (Mr 22,000) inhibitors were fairly specific for mammalian collagenase; the other unbound inhibitor was rather non-specific and also inhibited trypsin and thermolysin, but not bacterial collagenase. All the inhibitors were heat stable (90 degrees C, 30 min) and unaffected by 4-aminophenylmercuric acetate, but were inactivated by reduction and alkylation. PMID- 2556982 TI - Latent HSV-1 infection in mice immunized with a zwitterionic detergent-extracted HSV-1 antigen preparation. AB - A HSV-1 antigen preparation obtained by zwitterionic detergent-extraction of HSV 1 infected Vero cells was investigated for its ability to protect mice against establishment of latent infection in the trigeminal ganglion following HSV-1 challenge by ear scarification. The antigen preparation was shown to induce antibody production and correlation between pre-challenge antibody level and establishment of latency was found. Although immunization did not reduce the overall incidence of ear erythema following challenge the duration of erythema was significantly decreased. Latency in the trigeminal ganglion was also significantly reduced in vaccinated mice and in addition, a correlation was found between the duration of erythema and latency, there being significantly more latent infections in mice with erythema of three or more days duration, than in those with erythema lasting less than three days. PMID- 2556986 TI - The effects of cadmium on the p-nitrophenyl phosphatase and inorganic pyrophosphatase activities of alkaline phosphatase in developing hamster tooth germs. AB - p-Nitrophenyl phosphatase (p-NPP-ase) and inorganic pyrophosphatase (PPi-ase) activities originate from the same alkaline phosphatase enzyme. Only the PPi-ase site has zinc (Zn2+) as a cofactor. Cadmium (Cd2+) in concentrations from 10(-5) mol/l upwards inhibited the PPi-ase activity, but did not inhibit the p-NPP-ase activity at all. In mineralizing tooth germs Cd2+ may replace Zn2+, thereby changing the specific stereoconfiguration in the active centre needed for PPi-ase activity, but not that for p-NPP-ase activity. PMID- 2556987 TI - Interaction of streptococcal lipoteichoic acid with artificial tooth pellicle. AB - Artificial pellicles were prepared by coating hydroxyapatite beads with whole saliva. Radiolabelled lipoteichoic acid was isolated from Streptococcus sanguis NCTC 7863 by phenol extraction. Various concentrations of radiolabelled lipoteichoic acid were mixed with saliva-coated hydroxyapatite in the presence and absence of high ionic strength phosphate buffer, bovine serum albumin, gelatin, unlabelled lipoteichoic acid, Tween 20 and Triton X-100. The amount of lipoteichoic acid binding was measured by counting the residual radioactivity of the saliva-coated hydroxyapatite after thorough washing. In one experiment the binding was measured in the presence of a mucinous glycoprotein isolated from human saliva. The data were analysed by means of Scatchard and double reciprocal plots of the bound and unbound fraction of lipoteichoic acid. The lipoteichoic acid interacted hydrophobically with the saliva-coated hydroxyapatite; the interaction was complex with multiple binding sites exhibiting a range of affinities. The mean association constant was 1.1 x 10(9) M-1 and the minimum number of binding sites was 3.9 x 10(12)/mm2 of artificial pellicle. The salivary mucin competitively inhibited the interaction, which suggests that this may be one of the salivary components involved. PMID- 2556988 TI - Intraocular antiviral therapy. PMID- 2556989 TI - A controlled retrospective study of ganciclovir treatment for cytomegalovirus retinopathy. Use of a standardized system for the assessment of disease outcome. UCLA CMV Retinopathy. Study Group. AB - To evaluate more accurately the clinical response of cytomegalovirus retinopathy to ganciclovir, a system for the assessment of disease outcome was developed that uses retinal photographs and three factors: development of new lesions, enlargement of preexisting lesions, and change in retinal opacification of lesion borders. With this system a retrospective comparison was performed of 24 ganciclovir-treated patients and 17 untreated patients with cytomegalovirus retinopathy. A masked assessment showed disease progression in 10 treated patients (43%) during a median period of 22 days. In contrast, 16 untreated patients (94%) had progression of disease during a median period of 25 days. Comparison of treated and untreated eyes also suggests that treatment may prevent deterioration of visual acuity during the same period. This study supports the conclusions of previous uncontrolled studies that ganciclovir is beneficial in the treatment of patients with acquired immunodeficiency syndrome and cytomegalovirus retinopathy. It also demonstrates the utility of the proposed system for assessment of disease outcome that can be used in future studies of therapy for necrotizing viral retinopathies. PMID- 2556991 TI - Benign fibrous tumor of the choroid. AB - A 32-year-old white woman presented with an amelanotic choroidal tumor in the left eye. Although a malignant melanoma could not be totally ruled out, some features suggested a benign tumor. The patient was followed up without treatment during a 2-year period. Neovascular glaucoma developed and the blind eye was enucleated. Histopathologic examination revealed a benign fibrous tumor of the choroid, presumably a fibrous histiocytoma. PMID- 2556990 TI - Long-term intravitreal ganciclovir therapy for cytomegalovirus retinopathy. AB - The safety and efficacy of intravitreously administered ganciclovir sodium as sole treatment for cytomegalovirus retinitis complicating the acquired immunodeficiency syndrome was studied prospectively in seven patients. All but one of the patients had bilateral cytomegalovirus retinitis and none were able to tolerate therapy with systemically administered ganciclovir because of myelosuppression in six patients and hepatotoxicity in one patient. Intravitreal ganciclovir therapy was discontinued in two patients within the initial 2-week induction phase because of severe intractable thrombocytopenia in one patient and retinal detachment in the other. The retinal detachment could not be conclusively attributed to the injections and was probably a secondary complication of cytomegalovirus retinitis. The remaining five patients were treated weekly, with the course of therapy ranging from a minimum of 14 weeks (18 injections) to a maximum of 56 weeks (58 injections). The patients were followed up for an average of 23.5 weeks. All eyes responded to intravitreal therapy initially, while the six untreated control eyes with cytomegalovirus retinitis all demonstrated progression of disease. Two eyes relapsed while receiving intravitreal doses of 200 micrograms of ganciclovir sodium and were subsequently treated with 300 micrograms of ganciclovir sodium per injection. One eye responded to this regimen, while in the other one the disease progressed. In the long-term treatment group, one eye developed Staphylococcus epidermidis endophthalmitis, which was treated with vitrectomy and intravitreal and systemic antibiotics. PMID- 2556992 TI - Detection of CMP-N-acetylneuraminic acid hydroxylase activity in fractionated mouse liver. AB - The finding that N-glycoloylneuraminic acid (Neu5Gc) in pig submandibular gland is synthesized by hydroxylation of the sugar nucleotide CMP-Neu5Ac [Shaw & Schauer (1988) Biol. Chem. Hoppe-Seyler 369, 477-486] prompted us to investigate further the biosynthesis of this sialic acid in mouse liver. Free [14C]Neu5Ac, CMP-[14C]Neu5Ac and [14C]Neu5Ac glycosidically bound by Gal alpha 2-3- and Gal alpha 2-6-GlcNAc beta 1-4 linkages to fetuin were employed as potential substrates in experiments with fractionated mouse liver homogenates. The only substrate to be hydroxylated was the CMP-Neu5Ac glycoside. The product of the reaction was identified by chemical and enzymic methods as CMP-Neu5Gc. All of the CMP-Neu5Ac hydroxylase activity was detected in the high-speed supernatant fraction. The hydroxylase required a reduced nicotinamide nucleotide [NAD(P)H] coenzyme and molecular oxygen for activity. Furthermore, the activity of this enzyme was enhanced by exogenously added Fe2+ or Fe3+ ions, all other metal salts tested having a negligible or inhibitory influence. This hydroxylase is therefore tentatively classified as a monooxygenase. The cofactor requirement and CMP Neu5Ac substrate specificity are identical to those of the enzyme in high-speed supernatants of pig submandibular gland, suggesting that this is a common route of Neu5Gc biosynthesis. The relevance of these results to the regulation of Neu5Gc expression in sialoglycoconjugates is discussed. PMID- 2556993 TI - Spectroscopic studies of the type 2 and type 3 copper centres in the mercury derivative of laccase. AB - U.v.-visible-absorption and e.p.r. spectroscopy were used to study the type 2 and type 3 copper centres in the mercury derivative of laccase. After treatment with peroxide the mercury derivative of laccase exhibits a fully developed absorption band at 330 nm (delta epsilon = 2900 +/- 100 M-1.cm-1, which is characteristic of type 3 copper in the oxidized state. In addition, there is a weak ligand-field absorption at 740 nm (epsilon = 380 +/- 30 M-1.cm-1), which can be assigned to the type 3 pair. Because the e.p.r. spectrum of the type 2 copper is well resolved in the case of the mercury derivative of laccase, for the first time we have been able to observe spectroscopic evidence for a pH-dependent structural transition that has been invoked to explain the kinetics of enzyme reduction [Andreasson & Reinhammar (1979) Biochim. Biophys. Acta 568, 145-156]. According to the e.p.r. data the pKa lies in the range 6-7, and comparisons with a model compound show that the spectral changes can plausibly be interpreted in terms of the deprotonation of a water molecule in the co-ordination sphere of the type 2 copper. PMID- 2556994 TI - The purification and characterization of 4-ethylphenol methylenehydroxylase, a flavocytochrome from Pseudomonas putida JD1. AB - The enzyme 4-ethylphenol methylenehydroxylase was purified from Pseudomonas putida JD1 grown on 4-ethylphenol. It is a flavocytochrome c for which the Mr was found to be 120,000 by ultracentrifuging and 126,000 by gel filtration. The enzyme consists of two flavoprotein subunits each of Mr 50,000 and two cytochrome c subunits each of Mr 10,000. The redox potential of the cytochrome is 240 mV. Hydroxylation proceeds by dehydrogenation and hydration to give 1-(4' hydroxyphenyl)ethanol, which is also dehydrogenated by the same enzyme to 4 hydroxyacetophenone. The enzyme will hydroxylate p-cresol but is more active with alkylphenols with longer-chain alkyl groups. It is located in the periplasm of the bacterium. PMID- 2556995 TI - The quaternary structure in solution of human complement subcomponent C1r2C1s2. AB - C1r2C1s2 is a subcomponent of first component C1 of the complement cascade. Previously two distinct models for its structure have been described, in which C1r2C1s2 is either a linear rod-like assembly of the globular domains found in each of C1s and C1r, or these domains are arranged to form an asymmetric X-shaped structure. These two models were evaluated by using hydrodynamic simulations and neutron scattering. The data on C1s, C1s2 and C1r are readily represented by straight hydrodynamic cylinders, but not C1r2 or C1r2C1s2. Tests of the X structure for C1r2 and C1r2C1s2 successfully predicted the experimental sedimentation coefficients, thus supporting this model. Neutron scattering analyses on C1s and C1r2 are consistent with a linear structure for C1s, but not for C1r2. An X-shaped structure for C1r2 was found to give a good account of the neutron data at large scattering angles. The total length of the C1s and C1r monomers was determined as 17-20 nm, which is compatible with electron microscopy. On the basis of the known sequences of C1r and C1s, this length is accounted for by a linear arrangement of a serine-proteinase domain (length 4 nm), two short consensus repeat domains (2 x 4 nm), and a globular entity containing the I, II and III domains (4-7 nm). PMID- 2556996 TI - Indirect immunofluorescence localization of beta-adrenergic receptors and G proteins in human A431 cells. AB - Polyclonal antibodies directed against (i) rodent lung beta 2-adrenergic receptor, (ii) a synthetic fragment of an extracellular domain of the receptor, and (iii) human placenta G-protein beta-subunits, were used to localize these antigens in situ in intact and permeabilized human epidermoid carcinoma A431 cells. Antibodies directed against beta 2-adrenergic receptors showed a punctate immunofluorescence staining throughout the cell surface of fixed intact cells. Punctate staining was also observed in clones of Chinese hamster ovary cells transfected with an expression vector harbouring the gene for the hamster beta 2 adrenergic receptor. The immunofluorescence observed with anti-receptor antibodies paralleled the level of receptor expression. In contrast, the beta subunits common to G-proteins were not stained in fixed intact cells, presumably reflecting their intracellular localization. In detergent-permeabilized fixed cells, strong punctate staining of G beta-subunits was observed throughout the cytoplasm. This is the first indirect immunofluorescence localization of beta adrenergic receptors and G-proteins. Punctate immunofluorescence staining suggests that both antigens are distributed in clusters. PMID- 2556997 TI - Localization of beta-adrenergic receptors in A431 cells in situ. Effect of chronic exposure to agonist. AB - The status of beta-adrenergic receptors was investigated in A431 cells exposed to chronic stimulation by the beta-adrenergic agonist, (-)-isoproterenol. Specific binding of beta-adrenergic antagonist (-)-[125I]iodocyanopindolol declined to 60 80% below control values within 12 h of agonist treatment. This decline in ligand binding was also observed in high-speed membrane fractions prepared from agonist treated cells. Immunoblots probed with anti-receptor antibodies revealed both that beta-adrenergic receptors from untreated and treated cells migrated as 65,000-Mr peptides and that the cellular complement of receptor was unchanged. Indirect immunofluorescence localization of beta-adrenergic receptors was comparable in control (untreated) cells and cells challenged with (-) isoproterenol for 1, 12, or 24 h. Thus receptor complement, migration on SDS/polyacrylamide-gel electrophoresis, and localization in situ are largely unaffected by agonist stimulation. Receptor binding of antagonist radioligands, in contrast, is markedly down-regulated in cells stimulated chronically with beta adrenergic agonists. These data argue in favour of agonist-induced alteration(s) in the conformation of the receptor that preclude radioligand binding rather than agonist-induced receptor sequestration and/or degradation. PMID- 2556998 TI - Human fibroblasts release reactive oxygen species in response to interleukin-1 or tumour necrosis factor-alpha. AB - Human fibroblasts in primary culture released reactive oxygen species upon stimulation with cytokines such as interleukin-1 alpha (IL-1) or tumour necrosis factor-alpha (TNF). The primary radical produced was O2.- as determined by e.s.r. spin trapping and cytochrome c reduction. In contrast to the oxidative burst in granulocytes and monocytes, radical formation took place continuously for at least 4 h. Low-level chemiluminescence was increased by stimulation with IL-1 and TNF. Spectral characteristics and tests with azide led to the conclusion that the photoemissive species were excited carbonyls and not singlet oxygen. Further, there was a liberation of ethane from the cells. Radical production and light emission were not altered by either xanthine or allopurinol, nor by azide, cyanide or rotenone. O2.- production increased in the presence of NADH or NADPH, making an NAD(P)H oxidase a likely source. PMID- 2556999 TI - Purification and properties of cytochrome P-450-dependent 14 alpha-sterol demethylase from Candida albicans. AB - The purification of cytochrome P-450-dependent 14 alpha-sterol demethylase (P 450DM) from the important opportunistic fungal pathogen, Candida albicans, is described. Optimal purification (875-fold) was achieved by extracting the cytochrome from microsomes with sodium cholate followed by hydroxyapatite, octyl Sepharose and CM-Sepharose chromatographies, giving a cytochrome preparation of 17.5 nmol/mg of protein. By the use of SDS/polyacrylamide-gel electrophoresis the cytochrome was judged to be highly purified on the basis of Coomassie Brilliant Blue staining of protein. The Mr of P-450DM was estimated to be 51,000. The absorption spectrum of oxidized P-450DM was characteristic of a low-spin cytochrome, and its reduced CO complex had a Soret absorption peak at 447 nm. When reconstituted in a model membrane system of dilauroylphosphatidylcholine with NADPH and O2, P-450DM catalysed the complete 14 alpha-demethylation of lanosterol, which was inhibited by CO. The cytochrome appeared to have a high degree of substrate specificity; it was unable to oxidize a number of xenobiotic compounds in the reconstituted assay. PMID- 2557000 TI - Translocation to rat liver mitochondria of phosphatidate phosphohydrolase. AB - When a particle-free supernatant fraction from rat liver was incubated at 37 degrees C with mitochondria and oleate, some of the enzyme phosphatidate phosphohydrolase (PAP), initially present in the particle-free supernatant, was recovered, after the incubation, bound to mitochondria. This translocation of PAP from cytosol to mitochondria was stimulated by oleate or palmitate in a similar fashion to the stimulation of translocation of PAP to endoplasmic reticulum [Martin-Sanz, Hopewell & Brindley (1984) FEBS Lett. 175, 284-288]. Translocation of PAP from particle-free supernatant to a partially purified mitochondrial-outer membrane preparation was also stimulated by oleate. More PAP was bound to a mitochondrial-outer-membrane fraction washed in 0.5 M-NaCl before resuspension in sucrose than to a sucrose-washed mitochondrial-outer-membrane preparation. In contrast, washing of microsomal membranes in 0.5 M-NaCl did not enhance the binding of PAP to these membranes. PAP also binds to phosphatidate-loaded mitochondria or microsomes (microsomal fractions). In the experimental system employed, more PAP bound to mitochondria loaded with phosphatidate than to microsomes loaded with phosphatidate. The results are discussed in relation to the role of mitochondrial phosphatidate in liver lipid metabolism. PMID- 2557001 TI - The catalytic mechanism of the hydroxylation reaction of peptidyl proline and lysine does not require protein disulphide-isomerase activity. AB - Prolyl 4-hydroxylase, an alpha 2 beta 2 tetramer, catalyses the formation of 4 hydroxyproline in collagens. The beta subunit is known to be identical with the enzyme protein disulphide-isomerase and to possess disulphide-isomerase activity even when present in the prolyl 4-hydroxylase tetramer. We here report that lysyl hydroxylase, a homodimer, and algal prolyl 4-hydroxylase, a monomer, do not contain detectable protein disulphide-isomerase activity. Since the hydroxylase reaction mechanisms are similar, the data suggest that the protein disulphide isomerase activity of the vertebrate prolyl 4-hydroxylase beta subunit is unlikely to be involved in the catalytic mechanism of the hydroxylation reaction. PMID- 2557002 TI - Defective phosphorylation of a calmodulin-binding protein in cystic-fibrosis submandibular glands. AB - Calmodulin-binding proteins in fractions purified from human submandibular glands by calmodulin-Sepharose were phosphorylated with [gamma-32P]ATP, in the absence of exogenous protein kinase. The major proteins phosphorylated had molecular masses of 45, 51 and 61 kDa. Phosphorylation was increased by activators of protein kinase C and inhibited by H-7. Phosphorylation of the 61 kDa band was markedly decreased in cystic-fibrosis submandibular glands. PMID- 2557003 TI - Evidence for tight metabolic control of the receptor-activated polyphosphoinositide cycle in human platelets. AB - The [32P]PIP2/[32P]PA and the [32P]PIP/[32P]PA relationships were demonstrated to be remarkably similar after stimulation of [32P]Pi-prelabelled platelets for 90 s with various combinations and concentrations of agonists and inhibitors. Thus the activity of the PI and PIP kinases with the corresponding phosphomonoesterases may be tightly controlled during receptor-mediated platelet stimulation involving phospholipase C activation. PMID- 2557006 TI - Purification and properties of active atrial-natriuretic-peptide receptor (type C) from bovine lung. AB - Atrial-natriuretic-peptide (ANP) receptor, previously identified as a 140 kDa protein with a disulphide-linked homodimeric structure, was purified from bovine lung by (NH4)2SO4 fractionation and affinity chromatography on ANP-Affi-Gel 10. The purified receptor had a binding capacity of 4.2 nmol of ANP/mg of protein and an affinity constant of 6.5 pM. The isoelectric point of the receptor was 5.8, consistent with the acidic nature of the protein (amino acid analysis revealed a predominance of glutamic acid and aspartic acid residues). Treatment with endoglycosidase H and glycopeptidase F revealed that the receptor has three complex types of oligosaccharide chains per 70 kDa subunit. Deglycosylation of the receptor did not affect its binding activity. Reduction with dithiothreitol and reoxidation by dialysis revealed a strong tendency of the receptor subunits to dimerize via disulphide cross-linking; however, carboxymethylation of the reduced receptor indicated that the intersubunit disulphide bond is not necessary for the ligand-binding activity. PMID- 2557005 TI - Differential sensitivity of anti-IgM-induced and NaF-induced inositol phospholipid metabolism to serine protease inhibitors in BAL17 B lymphoma cells. AB - A BAL17 B lymphoma cell line bearing mu and delta chains on its surface behaves in a similar manner to normal mature B cells in terms of initial biochemical transmembrane signalling [Mizuguchi, Beaven, Ohara & Paul (1986) J. Immunol. 137, 2162-2167; Mizuguchi, Yong-Yong, Nakabayashi, Huang, Beaven, Chused & Paul (1987) J. Immunol. 139, 1054-1059]. Therefore the effects of protease inhibitors on increases in inositol phospholipid metabolism and intracellular free calcium concentration ([Ca2+]i) were examined. We show that the serine protease inhibitors Tos-Phe-CH2Cl (1-chloro-4-phenyl-3-L-tosylamidobutan-2-one-, TPCK) and Tos-Lys-CH2Cl (7-amino-1-chloro-3-L-tosylamidoheptan-2-one; TLCK) inhibit anti IgM-mediated accumulation of inositol phosphates in a dose-dependent manner. InsP3 production induced by anti-IgM is also inhibited by pretreatment with Tos Lys-CH2Cl or Tos-Phe-CH2Cl. Tos-Lys-CH2Cl- Tos-Phe-CH2Cl-mediated inhibition is not overcome by high concentrations of anti-IgM. Moreover, anti-IgM-mediated increases in [Ca2+]i are inhibited by pretreatment of the cells with these inhibitors. However, increases in inositol phospholipid metabolism caused by NaF, an activator of guanine-nucleotide-binding proteins (G-proteins), are approx. 10 fold more resistant to Tos-Lys-CH2Cl and Tos-Phe-CH2Cl inhibition compared with anti-IgM-induced changes. Furthermore, NaF-induced increases in [Ca2+]i are not inhibited by Tos-Lys-CH2Cl or Tos-Phe-CH2Cl pretreatment, suggesting that the inhibitors act at a step proximal to phospholipase C activation. The Tos-Lys CH2Cl or Tos-Phe-CH2Cl treatment does not change the membrane IgM density as measured by flow cytometry, indicating that the active site of the inhibitors is distal to the membrane IgM molecule. These results indicate that serine proteases may be involved in coupling the receptor cross-linkage to G-protein. PMID- 2557007 TI - Turnover of carnitine palmitoyltransferase mRNA and protein in H4IIE cells. Effect of cyclic AMP and insulin. AB - Regulation of the 68 kDa carnitine palmitoyltransferase (CPT) synthesis by (chlorophenylthio) cyclic AMP (cAMP) and insulin was studied in H4IIE cells in culture. Addition of 0.1 mM- or 1.0 mM-(chlorophenylthio) cAMP induced CPT mRNA and rate of transcription 2-4-fold by 15 min, reaching a plateau at 4-6-fold by 30 min. Addition of 5-15 nM-insulin plus 1.0 mM-cAMP suppressed the increases in transcription rate and mRNA levels occurring with cAMP alone. The t1/2 for CPT mRNA was 70-80 min and was not affected by cAMP. The t1/2 for CPT protein was 70 min, and was increased to 240 min in the presence of cAMP. The rate of CPT synthesis was also increased in the presence of cAMP. The data indicate that CPT synthesis is increased by cAMP via induction of transcription and subsequent increase in the CPT mRNA. Insulin acts to depress transcription and CPT mRNA. In addition, cAMP prolongs the t1/2 of CPT. PMID- 2557004 TI - High molecular mass intracellular proteases. PMID- 2557008 TI - Effects of oxyradicals on oxymyoglobin. Deoxygenation, haem removal and iron release. AB - We have examined the effects of O2-derived free radicals on oxymyoglobin, the myocardial intracellular protein involved in the storage and transport of O2. The oxyradicals generated by the xanthine/xanthine oxidase system decreased the concentration of oxymyoglobin. Based on the decreases in absorbance peaks at 581 nm and 415 nm it is estimated that out of a 10 nmol decrease in oxymyoglobin, 5 nmol appears to be oxidized to ferrimyoglobin (deoxygenation), while haem was removed from the other 5 nmol of haem protein. These processes were inhibited by both catalase alone and superoxide dismutase in combination with catalase, but not by either superoxide dismutase alone or deferoxamine. These results suggest that among H2O2, OH. and O2.-, only H2O2 causes the removal of haem and the oxidation of oxymyoglobin. Furthermore, the oxyradicals also released 3 microM free iron from oxymyoglobin, which is at least 5-fold less than the 15 nmol loss of oxymyoglobin. The loss of oxymyoglobin also preceded the release of free iron. These results indicate that oxymyoglobin oxidation and haem removal occur before the removal of free iron. Thus myoglobin appears to be highly susceptible to free radical attack, and this may represent yet another mechanism of free radical mediated cellular injury. PMID- 2557009 TI - Classification of prostaglandin receptors based on coupling to signal transduction systems. AB - A wide spectrum of prostaglandins (PG) stimulate both the production of cyclic AMP and an increase in free cytosolic Ca2+ concentration [( Ca2+]i) in the osteogenic osteosarcoma cell line, UMR-106-01, which has characteristics compatible with osteoblasts. Using PG-stimulated determinations of the second messengers cyclic AMP and [Ca2+]i, a method for classification of PG receptors is presented. UMR-106-01 cells demonstrate three subclasses of PG receptors. One receptor interacts with PGF2 alpha, PGD2, and thromboxane B2 (TxB2) to increase [Ca2+]i. A second receptor binds PGE2, PGE1, PGI2, PGA2 and 6-oxo-PGF1 alpha to increase [Ca2+]i by stimulation of a second separate phospholipase C pool. A third receptor accepts PGE2, PGE1, PGA2, PGI2 and to a lesser extent PGF2 alpha, PGD2 and TxB2 to increase cyclic AMP. Such a classification system may be applicable to other cells responding to multiple PGs by inducing changes in cellular second messengers. PMID- 2557011 TI - Stimulation of generation of inositol phosphates by carbamoylcholine and its inhibition by phorbol esters and iodide in dog thyroid cells. AB - The action of carbamoylcholine (Cchol), NaF and other agonists on the generation of inositol phosphates (IPs) was studied in dog thyroid slices prelabelled with myo-[2-3H]inositol. The stimulation by Cchol (0.1 microM-0.1 mM) of IPs accumulation through activation of a muscarinic receptor [Graff, Mockel, Laurent, Erneux & Dumont (1987) FEBS Lett. 210, 204-210] was pertussis- and cholera-toxin insensitive. Ins(1,4,5)P3, Ins(1,3,4)P3 and InsP4 were generated. NaF (5-20 mM) also increased IPs generation (Graff et al., 1987); this effect was potentiated by AlCl3 (10 microM) and unaffected by pertussis toxin. Although phorbol dibutyrate (5 microM) abolished the cholinergic stimulation of IPs generation (Graff et al., 1987), it did not affect the fluoride-induced response. Cchol and NaF did not require extracellular Ca2+ to exert their effect, and neither KCl induced membrane depolarization nor ionophore A23187 (10 microM) had any influence on basal IPs levels, or on cholinergic stimulation. However, more stringent Ca2+ depletion with EGTA (0.1 or 1 mM) decreased basal IPs levels as well as the amplitude of the stimulation by Cchol without abolishing it. Dibutyryl cyclic AMP, forskolin, cholera toxin and prostaglandin E1 had no effect on basal IPs levels and did not decrease the response to Cchol. Iodide (4 or 40 microM) also strongly decreased the cholinergic action on IPs, this inhibition being relieved by methimazole (1 mM). Our data suggest that Cchol activates a phospholipase C hydrolysing PtdIns(4,5)P2 in the dog thyroid cell in a cyclic AMP independent manner. This activation requires no extracellular Ca2+ and depends on a GTP-binding protein insensitive to both cholera toxin and requires no extracellular Ca2+ and depends on a GTP-binding protein insensitive to both cholera toxin and pertussis toxin. The data are consistent with a rapid metabolism of Ins(1,4,5)P3 to Ins(1,3,4)P3 via the Ins(1,4,5)P3 3-kinase pathway, followed by dephosphorylation by a 5-phosphomonoesterase. Indeed, a Ca2+ sensitive InsP3 3-kinase activity was demonstrated in tissue homogenate. Stimulation of protein kinase C and an organified form of iodine inhibit the Cchol-induced IPs generation. The negative feedback of activated protein kinase C could be exerted at the level of the receptor or of the receptor-G-protein interaction. PMID- 2557010 TI - Co-ordinated changes in the cyclic AMP signalling system and the phosphorylation of two nuclear proteins of Mr 130,000 and 110,000 during proliferative stimulation of the rat parotid gland by isoprenaline. Possible identity of the two proteins with pp135 and nucleolin. AB - Parotid glands were stimulated to growth by repeated injection of the beta agonist isoprenaline into rats. Incubation of intact parotid-gland lobules with [32P]Pi and subsequent analysis of nuclear proteins revealed in the stimulated glands an increased 32P incorporation into two acid-soluble non-histone proteins with apparent Mr values of 110,000 and 130,000 (p110 and p130). After a single injection of isoprenaline, leading to a biphasic increase in DNA synthesis (maximum at 24 h), the same two proteins showed a transiently increased 32P incorporation at 17 h after injection. At this time point at the onset of DNA synthesis the total activity of soluble cyclic AMP-dependent protein kinase decreased. No change in p110/p130 phosphorylation was observed at 0.3 h after stimulation, a time of maximal stimulation of secretion. Administration of the beta-antagonist propranolol 8 h after the injection of isoprenaline suppressed the increase in DNA synthesis, the preceding changes in the concentration of cyclic AMP and in the activity of cyclic AMP-dependent protein kinase, as well as the increased phosphorylation of p110 and p130. Cross-reactivity of p110 and p130 with specific antisera against two nucleolar phosphoproteins of similar molecular mass (nucleolin and pp135), as well as their localization in a nucleolar cell fraction, indicated a possible identity of p110 and p130 with these two proteins. Our results suggest that nucleolin and pp135 are nuclear target proteins of cyclic AMP in the cyclic AMP-influenced regulation of the transition of cells from the G1 to the S phase. PMID- 2557013 TI - Influence of superoxide on myeloperoxidase kinetics measured with a hydrogen peroxide electrode. AB - Stimulated neutrophils discharge large quantities of superoxide (O2.-), which dismutates to form H2O2. In combination with Cl-, H2O2 is converted into the potent oxidant hypochlorous acid (HOCl) by the haem enzyme myeloperoxidase. We have used an H2O2 electrode to monitor H2O2 uptake by myeloperoxidase, and have shown that in the presence of Cl- this accurately represents production of HOCl. Monochlorodimedon, which is routinely used to assay production of HOCl, inhibited H2O2 uptake by 95%. This result confirms that monochlorodimedon inhibits myeloperoxidase, and that the monochlorodimedon assay grossly underestimates the activity of myeloperoxidase. With 10 microM-H2O2 and 100 mM-Cl-, myeloperoxidase had a neutral pH optimum. Increasing the H2O2 concentration to 100 microM lowered the pH optimum to pH 6.5. Above the pH optimum there was a burst of H2O2 uptake that rapidly declined due to accumulation of Compound II. High concentrations of H2O2 inhibited myeloperoxidase and promoted the formation of Compound II. These effects of H2O2 were decreased at higher concentrations of Cl-. We propose that H2O2 competes with Cl- for Compound I and reduces it to Compound II, thereby inhibiting myeloperoxidase. Above pH 6.5, O2.- generated by xanthine oxidase and acetaldehyde prevented H2O2 from inhibiting myeloperoxidase, increasing the initial rate of H2O2 uptake. O2.- allowed myeloperoxidase to function optimally with 100 microM-H2O2 at pH 7.0. This occurred because, as previously demonstrated, O2.- prevents Compound II from accumulating by reducing it to ferric myeloperoxidase. In contrast, at pH 6.0, where Compound II did not accumulate, O2.- retarded the uptake of H2O2. We propose that by generating O2.- neutrophils prevent H2O2 and other one-electron donors from inhibiting myeloperoxidase, and ensure that this enzyme functions optimally at neutral pH. PMID- 2557012 TI - Effect of basic polycations and proteins on purified insulin receptor. Insulin independent activation of the receptor tyrosine-specific protein kinase by poly(L lysine). AB - Since the studies on tyrosine phosphorylation of calmodulin by the insulin receptor kinase in vitro suggested that protamine and poly(L-lysine) may activate phosphorylation of the receptor beta subunit [Sacks & McDonald (1988) J. Biol. Chem. 263, 2377-2383], we examined the effects of a variety of basic polycations/proteins and polyamines on insulin receptor kinase activity. The insulin receptor purified from human placental membranes was incubated with each basic polycation/protein or polyamine and assayed for tyrosine-specific protein kinase activity by measuring 32P incorporation into the src-related peptide. At a concentration of 1 microM, poly(L-lysine) and poly(L-ornithine) markedly stimulated kinase activity, whereas poly(L-arginine) and histones H1 and H2B inhibited insulin receptor kinase. In contrast, at a concentration of 1 mM, three polyamines (spermine, spermidine and putrescine) did not alter kinase activity. Poly(L-lysine) and poly(L-ornithine) stimulated the insulin receptor kinase by 5 10-fold at concentrations of 0.1-1 microM. Protamine sulphate also showed a significant stimulatory effect at a concentration of 100 microM. Preincubation of the receptor with poly(L-lysine) or poly(L-ornithine) for 20-60 min resulted in maximal kinase activation. Poly(L-lysine), the most effective activator of the receptor kinase, was used to characterize further the mechanisms of the kinase activation. Poly(L-lysine) activates the insulin receptor kinase by increasing the Vmax. without changing the Km. Poly(L-lysine) markedly stimulates the kinase activity of insulin receptor preparations that have lost both basal kinase activity and the ability to be stimulated by insulin. Insulin and poly(L-lysine) also differed in their ability to stimulate the kinase activity of prephosphorylated receptors. Prephosphorylation of the receptors did not affect the stimulation of the kinase by insulin. In contrast, prephosphorylation of receptors resulted in a markedly enhanced ability of poly(L-lysine) to stimulate kinase activity. These studies suggest that the mechanisms by which poly(L lysine) and insulin activate the kinase are different. In conjunction with other additional evidence, it is suggested that poly(L-lysine) interacts directly with the beta-subunit of the receptor, thereby activating the receptor kinase. PMID- 2557014 TI - Adaptative decrease in expression of the mRNA for uncoupling protein and subunit II of cytochrome c oxidase in rat brown adipose tissue during pregnancy and lactation. AB - Uncoupling-protein (UCP) mRNA expression is decreased to 15% of virgin control levels between days 10 and 15 of pregnancy, and remains at these low values during late pregnancy and lactation. Abrupt weaning of mid-lactating rats causes a slight but significant increase in UCP mRNA. Expression of mRNA for subunit II of cytochrome c oxidase (COII) decreased to half that of virgin control in late pregnancy and during lactation. Whereas COII mRNA expression is in step with the known modifications of brown-fat mitochondria content during the breeding cycle of the rat, UCP mRNA expression appears to be diminished much earlier than the mitochondrial proton-conductance-pathway activity. On the other hand, the reactivity of brown fat to increase expression of UCP and COII mRNAs in response to acute cold or noradrenaline treatment is not impaired during lactation. PMID- 2557015 TI - Endothelin stimulates DNA synthesis in Swiss 3T3 cells. Synergy with polypeptide growth factors. AB - The vasoactive peptide endothelin is shown to be a potent mitogen for Swiss 3T3 cells. Although endothelin has little effect on DNA synthesis when added alone to cells in serum-free medium, the peptide synergizes very strongly with several other growth factors. A half-maximal response to endothelin is observed at approx. 0.3 nM, with a maximal effect at 3 nM. Over the same concentration range, endothelin stimulates a 2-fold increase in the accumulation of cellular inositol phosphates. Endothelin may prove to be a useful additional agonist for studying the signalling pathways involved in the control of 3T3-cell proliferation. PMID- 2557016 TI - Benzodiazepine-like molecules, as well as other ligands for the brain benzodiazepine receptors, are relatively common constituents of plants. AB - The presence of benzodiazepine (BZD)-like molecules as well as of other substances with affinity for the brain BZD-receptors was explored in eight non flowering plants known to contain biflavonoids, three flowering plants used as sedatives in folkloric medicine and one plant extensively used in Argentina, Uruguay, Brazil and Paraguay as a tea substitute. All the plants examined contained substances which bound to the central BZD-receptors and the majority of them also had BZD-like compounds detected by their specific interaction with a monoclonal antibody against BZDs. In various cases this last type of compound was present in amounts which exceeded trace levels (0.5-1.0 ng/g). The biological or clinical significance for humans of all these substances should be explored. PMID- 2557017 TI - Cholera toxin ADP-ribosylates the receptor-coupled form of pertussis toxin sensitive G-proteins. AB - Cholera toxin catalyzes the ADP-ribosylation of 40 kDa pertussis toxin substrates in membranes from NG108-15 cells, which is increased in the presence of the opioid agonist DADLE. The basal ADP-ribosylation can be abolished by the opioid antagonist ICI 174864, suggesting that unoccupied opioid receptors interact spontaneously with the pertussis toxin substrates Gi/Go in the membrane. Treatment of NG108-15 cells with the opioid agonist DADLE leads to a reduction of agonist-stimulated and basal ADP-ribosylation of 40 kDa substrates catalyzed by cholera toxin. This indicates that the spontaneous interaction between opioid receptors and G-proteins is decreased in membranes of cells in which the receptor was desensitized by prolonged exposure to the agonist. PMID- 2557019 TI - Sugar modified oligonucleotides. I. Carbo-oligodeoxynucleotides as potential antisense agents. AB - For the first time, carbo-oligodeoxynucleotides, namely c-dT4 and c-dT12, have been synthesized. As compared to the natural oligomers these carbo oligodeoxynucleotides are at least 5 times more stable toward enzymatic degradation and bind more strongly to complementary DNA. These preliminary data indicate that such oligomers fulfill the requirements to be considered as potential antisense agents. PMID- 2557018 TI - Cyclic GMP analogs inhibit gamma thrombin-induced arachidonic acid release in human platelets. AB - Elevation in intracellular cyclic GMP levels is the proposed proximal mechanism for the vasodilatory and platelet inhibitory action of nitrovasodilators and of nitric oxide, the putative endothelium-derived relaxing factor. In this study, the stable cyclic GMP analogs, 8-bromo-cGMP and N2, 2'-O-dibutyryl-cGMP were found to inhibit the release of [3H]-arachidonic acid from gamma thrombin stimulated human platelets in a time- and dose-dependent manner. Inhibition of the formation of arachidonic acid metabolites, 12-HETE and thromboxane B2, paralleled that of arachidonic acid release and was accompanied by a dose dependent inhibition of platelet aggregation. The formation of phosphatidic acid, a metabolite of phospholipase C, however, was relatively preserved. At a concentration of 8-bromo-cGMP (2 mM) that produced near-total inhibition of arachidonic acid release, phosphatidic acid formation remained at 60% of control levels. Thus, cGMP analogs have a preferential inhibitory effect on the release and subsequent metabolism of arachidonic acid. The phospholipase A2/arachidonic acid pathway appears to be an important target for the physiologic action of cGMP, and EDRF, and for the pharmacologic action of nitrovasodilators. PMID- 2557020 TI - Alterations in the creatine kinase system in the myocardium of cardiomyopathic hamsters. AB - Immunochemical and biochemical methods were used to assess quantitatively the changes in the heart creatine kinase system in the myopathic Syrian hamsters, line CHF I46. Cardiomyopathy in I75-200 day old animals was characterized by decreased content of mitochondria and lower total creatine kinase activity. In isolated mitochondria only the creatine kinase activity was decreased while cytochromes aa3 content and respiration rate were unchanged. The share of mitochondrial creatine kinase in the total tissue enzyme activity was decreased from 33% to I8% and that of BB form was elevated from 5% in control to 20%, at unchanged relative level of MM. Immunoassay showed decreased amount of the mitochondrial creatine kinase in the tissue and its decreased ratio to cytochromes aa3. The results show altered expression of creatine kinase isoenzymes in cardiomyopathy. PMID- 2557021 TI - Histamine stimulates calcium-mediated protein phosphorylation in a colonic epithelial cell line. AB - Protein phosphorylation responses in intact enterocytes were examined by stimulating 32Pi-labeled T84 cell monolayers with histamine and resolving proteins by two-dimensional gel electrophoresis. Histamine increases 32P incorporation into two acidic proteins of Mr 83,000 and of Mr 29,000, designated p83 and p29. Labeling of p83 and p29 is also increased in cells exposed to ionomycin, but not in cells exposed to vasoactive intestinal peptide under conditions resulting in cAMP-mediated secretion and cAMP-stimulated protein phosphorylation. When T84 cell fractions are incubated with [gamma-32P]ATP, labeling of p83 is stimulated by Ca++, but not by cAMP. Thus, histamine stimulates Ca++-mediated protein phosphorylation during the regulation of Cl- secretion. PMID- 2557022 TI - Separate regulatory control of apical and basolateral Na+/H+ exchange in renal epithelial cells. AB - Epithelial layers of LLC-PK1/PKE20 cells, a renal epithelial cell line which expresses Na+/H+ exchange activities in the apical as well as basolateral membrane domains, are examined in the single cell mode by microspectrofluorometry. We provide evidence that basolateral Na+/H+ exchange is more sensitive to amiloride inhibition than is apical Na+/H+ exchange. Furthermore, we demonstrate that the two exchange activities differ in their regulatory control: kinase A activation (forskolin, 8-Br-cAMP) leads to inhibition of both exchange activities, whereas kinase C activation (phorbol ester) stimulates basolateral and inhibits apical Na+/H+ exchange. Thus, renal epithelial cells may contain two Na+/H+ exchange activities: an apical ("epithelial") and basolateral ("housekeeping") which may serve different cellular functions and are under separate regulatory controls. PMID- 2557023 TI - Proteolytic conversion of xanthine dehydrogenase to xanthine oxidase: evidence against a role for calcium-activated protease (calpain). AB - The present study tested the hypothesis that calpain is responsible for the limited proteolytic conversion of xanthine dehydrogenase (XD) to xanthine oxidase (XO). We compared the effects of various proteases on the activity and molecular weight of a purified preparation of xanthine dehydrogenase from rat liver. In agreement with previous reports, trypsin treatment produced a complete conversion of XD to XO accompanied by a limited proteolysis of XDH from an Mr of 140 kD to an Mr of 90 kD. Treatment with calpain I or calpain II did not produce a conversion from XD to XO nor did it result in partial proteolysis of the enzyme. Similarly, trypsin treatment partially degraded a reversibly oxidized form of xanthine dehydrogenase while calpain I or calpain II were ineffective. The possibility that an endogenous inhibitor prevented the proteolysis of XDH by calpain I or II was excluded by verifying that brain spectrin, a known calpain substrate, was degraded under the same incubation conditions. The results indicate that calpain is not likely to be responsible for the in vivo conversion of XD to XO under pathological conditions. PMID- 2557024 TI - Decreased incorporation of glucose into lipids and increased lactate production by adipose tissue after long-term treatment of rats with D-fenfluramine. AB - Male rats were treated with ten daily doses of 10 mg of D-fenfluramine/kg. Body weight decreased after days 1 and 2, but thereafter the weight gain paralleled that of the control rats. After the tenth injection there were decreases in the weights of the epididymal fat pads, their fat content, and the average size of the adipocytes after collaginase digestion. The rate of glucose uptake by incubated pieces of adipose tissue was maintained after D-fenfluramine treatment, and the production of lactate increased. The incorporation of glucose into fatty acids by adipose tissue pieces decreased by 65-74% after treatment with D fenfluramine. This effect was not reversed by adding insulin or phenylisopropyladenosine to the incubations. D-Fenfluramine also decreased the incorporation of glucose into glyceride-glycerol, but this effect was less pronounced than that for fatty acid synthesis. Direct addition of D-fenfluramine to the incubation inhibited lipid synthesis from [14C]glucose but only at drug concentrations above 1 mM. It is concluded that the treatment of rats with D fenfluramine modifies the metabolic balance of adipose tissue so as to direct glucose metabolism away from lipid synthesis and towards lactate production. This could be a significant mechanism in the overall loss of adipose tissue mass caused by the administration of D-fenfluramine. PMID- 2557025 TI - The effect of some antiprostatic steroids upon cortisol production by guinea-pig adrenal cells stimulated by ACTH. AB - The antiprostatic steroids 6-methylene-4-pregnene-3,20-dione (6-MP) (I), 17-alpha acetoxy-6, 16-dimethylene-4-pregnene-3,20-dione (II), and melengestrol acetate (MGA) (III) were incubated with guinea-pig adrenal cells, both alone and maximally stimulated with ACTH. Cortisol output was then measured by RIA. Increased cortisol-like secretion was obtained with 6-MP in the absence of ACTH. In the presence of ACTH, cortisol-like steroid secretion was the sum of that seen with ACTH and 6-MP alone. It follows that 6-MP stimulates in vitro a cortisol like steroid cross reacting with the cortisol antibody by a mechanism that by passes ACTH. Steroid (II) weakly inhibited cortisol output. MGA, in contrast, proved to be a strong inhibitor of cortisol output (ID50 of 2.3 mumol/l). Its site of action was established by adding it to adrenal cells incubated with precursor steroids on the cortisol pathway. Conversion of 3 beta-hydroxysteroids to cortisol was inhibited whereas conversion of 3-keto steroids was not affected. It follows that MGA inhibits 3 beta-hydroxysteroid dehydrogenase. PMID- 2557026 TI - Comparative ability of digoxin and an aminosugar cardiac glycoside to bind to and inhibit Na+,K+-adenosine triphosphatase. Effect of potassium. AB - We compared the abilities of digoxin and aminogalactose digitoxigenin (ASI-222) to bind to, or inhibit, purified dog heart Na+,K+-ATPase in the presence of 1, 10, or 80 mM potassium chloride. Changing the potassium concentration from 1 to 10 mM increased the dose producing 50% inhibition of enzyme activity (IC50) by 9- and 2.5-fold for digoxin and ASI-222 respectively. Raising the potassium concentration to 80 mM increased the IC50 for digoxin 3-fold but did not alter significantly the IC50 for ASI-222. Equilibrium binding studies showed that this enzyme exhibited a single class of specific binding sites for both digoxin and ASI-222. Raising the potassium concentration did not affect the maximum number of binding sites (Bmax) but increased the apparent dissociation constant (KD) for digoxin. Potassium differentially affected the affinity and number of binding sites for ASI-222; raising the potassium concentration from 1 to 10 mM did not affect the Bmax or the KD, but raising it to 80 mM increased both. The effect of i.v. infusion of potassium chloride upon cardiac upon cardiac arrhythmias produced by i.v. infusion of digoxin or ASI-222 in anesthetized dogs was also determined. Infusion of potassium chloride reversed the cardiac arrhymias due to digoxin to normal rhythm, but not those due to ASI-222. In conclusion, the interaction of digoxin and the polar digitalis agent, ASI-222, with dog heart Na+,K+-ATPase was differentially affected by potassium. These agents also also produced cardiac arrhythmias, which were differentially affected by potassium. PMID- 2557027 TI - Comparison of intracellular drug retention, DNA damage and cytotoxicity of derivatives of doxorubicin and daunorubicin in a human colon adenocarcinoma cell line (LoVo). AB - Formation of DNA single strand breaks (SSB) was assayed by alkaline elution in LoVo cells treated with doxorubicin, daunorubicin and six derivatives of these drugs modified either in the chromophore or the sugar. Seven compounds showed a biphasic relationship (initial increase and then a decrease) for the formation of DNA-SSB over the concentration range 0.05-10 micrograms/ml. At a drug concentration in the range causing an increase of DNA damage very fast repair of DNA-SSB was observed for 4'-deoxydoxorubicin and 4-demethoxydaunorubicin; the kinetics of DNA-SSB investigated after drug removal at a drug concentration reducing DNA-SSB showed a time dependent increase of DNA damage for both drugs although with different patterns. 4'-Deoxydoxorubicin reduced the effect of radiations on the rate of elution of DNA in a way resembling the formation of DNA interstrand cross links (ISC) at concentrations at which DNA-SSB were reduced. DNA-ISC were not produced by chemical reactions occurring during sample processing for alkaline elution and this derivative was not metabolized by LoVo cells. The IC50 of the anthracyclines were on a several log range, though for most of the derivatives the cytotoxicity curve showed a plateau at growth inhibition of about 15-30% at increasing intracellular drug levels. A relationship between DNA damage and cytotoxicity was observed only in a very small range of DNA-SSB. It is likely that the different effects of these anthracyclines on the formation of DNA-SSB depend on a qualitatively different interaction between drug-DNA and topoisomerase II when the drug concentration is raised. PMID- 2557028 TI - Growth, differentiation and the beta-adrenergic signal system of HL-60 cells. Characterization in a medium with insulin as the only added protein. AB - The purpose of the present investigation was to define experimental conditions for studies on growth, differentiation and the beta-adrenergic signal system of HL-60 cells. The cell medium was completely devoid of added proteins and hormones other than insulin. The HL-60 cell was able to grow and differentiate in this medium. The spontaneous differentiation along the granulocytic pathway after 72 hr, as assessed by the Nitro Blue tetrazolium test, increased by 400% compared to the serum supplemented medium, but the response to 1 microM retinoic acid was equal in the two media. Induction of monocytic differentiation by 0.16 microM phorbol-13-acetate-12-myristate, as determined by surface adherence after 24 hr, was lower in the absence than in the presence of serum. cAMP levels were elevated in response to (-)-isoproterenol. The EC50 was 0.36 +/- 0.01 microM (mean +/- SE, N = 3). The beta-adrenergic ligand 3H-CGP 12177 was specifically bound to 1 single class of binding sites (Kd: 0.15 +/- 0.04 nM, Bmax: 2220 +/- 150, mean +/- SE, N = 3). These data are comparable to our previously reported findings in serum supplemented medium. The present data show that HL-60 cells are able to grow and differentiate in the absence of serum proteins and hormones other than insulin. Under the present experimental conditions, these cells possessed functional beta-adrenergic receptors. PMID- 2557029 TI - Microsomal superoxide anion production and NADPH-oxidation in a series of 22 aziridinylbenzoquinones. AB - Several 2,5-bis(1-aziridinyl)-1,4-benzoquinones (BABQs) can be activated to alkylating species by reduction of the quinone moiety. On the other hand, cytotoxicity of these compounds can be induced by redox cycling. A series of BABQs and their methylated analogues (BMABQs) with different substituents at the 3- and 6-position was synthesized in order to investigate the influence of the substituents on the reduction of the quinone moiety and on the generation of superoxide anion radicals with rat liver microsomes. Superoxide anion production (SAP) ranged from 3.7 +/- 0.1 to 742 +/- 74 nmoles/min/mg protein with quinone concentrations of 10 nmoles/ml. NADPH-oxidation was measured under the same conditions and it correlated well (r = 0.88, P less than 0.001) with SAP. It ranged from 1.4 +/- 0.2 to 494 +/- 60 nmoles/min/mg protein. SAP for 22 B(M) ABQs showed a good correlation with the summated electronic substituent constant sigma para.total (r = 0.86, P less than 0.001). It can be concluded that superoxide anion production by 22 B(M)ABQs in rat liver microsomes can be predicted from structural features of the compounds. PMID- 2557030 TI - Chronic benzodiazepine administration. IV. Rapid development of tolerance and receptor downregulation associated with alprazolam administration. AB - The triazolobenzodiazepine compound alprazolam may have unique clinical effects compared to other benzodiazepines, and both behavioral and neurochemical studies have indicated unusual results after acute doses of alprazolam. To determine the effects of chronic dosage in mice, alprazolam (2 mg/kg/day) was administered via osmotic pumps for 1-14 days, and open-field activity, plasma and brain concentrations, benzodiazepine receptor binding in vivo and in vitro, [35S]t butylbicyclophosphorothionate ([35S]TBPS) binding, and muscimol-stimulated chloride uptake were determined. Alprazolam decreased motor activity after 1 and 2 days, but tolerance developed by day 4 and persisted to day 14. Plasma and brain concentrations remained constant during the 2-week period. Benzodiazepine receptor binding in vivo was decreased at day 4 compared to day 1 in cortex (CX) and hypothalamus (HYPO), and remained depressed to day 14 in CX but not HYPO. Benzodiazepine binding in vitro and [35S]TBPS binding were decreased in CX at day 7. Muscimol-stimulated [36Cl-] uptake was decreased at days 4 and 7 compared to day 1, but at day 14 uptake was similar to day 1. These results indicate that behavioral tolerance and receptor downregulation develop rapidly during chronic alprazolam administration. Behavioral and neurochemical changes were similar to those associated with lorazepam administration, but occurred more rapidly and with different regional specificity. PMID- 2557031 TI - Species differences and heterogeneity of solubilized peripheral-type benzodiazepine binding sites. AB - The pharmacological characteristics of digitonin-solubilized peripheral-type benzodiazepine binding sites (PBS) from kidney membranes of various species were investigated to determine whether the species differences and heterogeneity observed in membrane-bound binding sites would be maintained after solubilization. [3H]PK 11195 (0.05 to 10 nM) bound with high affinity to rat, guinea pig, calf, and cat kidney solubilized preparations yielding maximal numbers of binding sites (Bmax) of 3,593 +/- 381, 25,645 +/- 1,795, 1,327 +/- 141, and 2,446 +/- 148 fmol/mg protein, respectively, and equilibrium dissociation constant (KD) values of 1.74 +/- 0.18, 2.15 +/- 0.15, 0.85 +/- 0.09, and 1.02 +/- 0.06 nM, respectively. On the other hand, the respective Bmax and KD values for [3H]Ro 5-4864 (1.25 to 40 nM) were 2,688 +/- 275, 14,182 +/- 1,134, 144 +/- 23 and 205 +/- 17 fmol/mg protein (about 75, 55, 11, and 8%, respectively, of that of [3H]PK 11195) and 13.8 +/- 1.5, 14.6 +/- 1.1, 10.6 +/- 1.7, and 19.9 +/- 1.2 nM. Unlabeled Ro 5-4864 was two orders of magnitude more potent in displacing [3H]PK 11195 binding from rat kidney solubilized preparations than from calf kidney solubilized preparations, whereas the potency of unlabeled PK 11195 in displacing [3H]PK 11195 binding from both rat and calf kidney solubilized preparations was almost identical. Analysis of these displacement data revealed that PK 11195 bound to a single population of binding sites (nH approximately equal to 1.0), whereas Ro 5-4864 bound to two populations of binding sites (nH less than 1.0) in both rat and calf kidney solubilized preparations. These results indicate that PBS species differences and heterogeneity observed in membrane-bound binding sites are retained in the soluble state and are probably attributable to variations in the molecular structure of PBS rather than to differences in membrane environment. PMID- 2557032 TI - Sodium regulation of alpha 2-adrenoreceptors in Dahl rats. Effect of feeding a low or high salt diet. AB - Sodium regulation of alpha 2-adrenoreceptors was investigated in inbred salt sensitive (S) and inbred salt-resistant (R) rats fed a high or low salt diet. The systolic blood pressure was higher in S rats than in R rats, and this difference was obviously greater on a high salt diet. In rats fed a low or high salt diet, S rats had higher alpha 2-adrenoreceptor density in the kidneys compared with R rats as measured by [3H]yohimbine binding and Scatchard analysis. The affinity of the receptors in the kidney for the antagonist, yohimbine, was nearly the same in these two strains either on a low or high salt diet. In the brain, the affinities or the numbers of receptors were not significantly different whether these two strains were fed a low or high salt diet. Inclusion of NaCl up to 80 mM in the assay medium did not alter the in vitro binding of [3H]yohimbine in the kidney or brain. On the other hand, inclusion of NaCl in the assay medium reduced the ability of epinephrine in competing with [3H]yohimbine for the receptor sites in the kidney and in the brain, and this effect of NaCl was the same in a given tissue between S and R rats, whether they were fed a low or high salt diet. These results suggest that: (1) in the kidneys, the receptor density and not the receptor affinity was different between S and R strains whether they were fed a low or high salt diet; (2) in the brain, the receptor density and affinity were the same between S and R rats regardless of the diet (low or high salt), indicating that the sodium salt diet modulates the peripheral but not the central alpha 2-adrenoreceptors; and this modulatory effect was observed only in S rats; (3) Na+ was able to reduce the affinity of the agonist (epinephrine) for the receptors in both S and R rats, and this effect of Na+ on central and peripheral alpha 2-adrenoreceptors was similar in prehypertensive rats and rats with salt induced hypertension; and (4) the resistance of R rats to salt-induced hypertension was not due to the absence of Na+ binding component involved in the regulation of alpha 2-adrenoreceptor-adenylate cyclase complex. PMID- 2557034 TI - Mechanistic aspects of paraquat toxicity in E. coli. A spin trapping study. AB - Mechanistic aspects of paraquat monocation radical (PQ.+) and copper involvement in paraquat toxicity have been examined using E. coli B cells. Electron spin resonance (ESR) spectrometry combined with cell survival studies were used to explore the correlation between radical production and biological damage. The line broadening agent oxalato-chromiate (CrOx) was used to characterize the anoxic partition of PQ.+ inside and outside the cell. In the presence of CrOx the ESR signal was totally eliminated, indicating that intracellular species were undetectable and that, contrary to previous reports, PQ.+ exclusively accumulates outside the cell. The PQ.+ radical does not react with H2O2 but disappears in the presence of H2O2 when catalytic traces of Cu(II) are present. Spin-trapping studies using DMPO showed that in aerobic environment paraquat-induced O2 radicals are detectable exclusively in the extracellular compartment. The correlation between PQ.+ appearance and the biological damage is not simple. PQ.+ non-toxically accumulates, in the absence of oxygen and either Cu(II) or H2O2. By contrast, with both H2O2 and Cu(II) the cells are rapidly killed but PQ.+ was undetectable. These results reconfirm the key catalytic mediatory function of transition metals in paraquat toxicity. PMID- 2557033 TI - Effect of verapamil on phosphate-induced changes in oxidative phosphorylation and atractyloside-sensitive adenine nucleotide translocase activity in two populations of rat heart mitochondria. AB - Phosphate (Pi)-induced depression in cardiac mitochondrial function was studied using mitochondria isolated by two different procedures which purportedly yield two distinct populations. Subsarcolemmal mitochondria (SLM) exhibited an enhanced sensitivity to 20 mM Pi with respect to oxidative phosphorylation. Thus, a significant depression in oxidative phosphorylation in this population was seen following only 1-min treatment, whereas interfibrillar mitochondria (IFM) were unaffected. Both populations showed a similar response to 5-min treatment with Pi. The Pi-induced depression in respiration was partially, although significantly, reversed by a 50 microM concentration of the calcium antagonist verapamil, an observation which suggests a contribution of calcium to the Pi induced defect in respiration. Pi also produced a potent inhibition of ADP uptake in both mitochondrial populations, which was in close agreement to Pi-induced modification of low amplitude shrinkage-swelling responses following ADP addition. Both of these parameters were unaffected by verapamil. Our results show an enhanced sensitivity of SLM to a verapamil-sensitive Pi-induced depression in oxidative phosphorylation. However, the potent, verapamil-insensitive decrease in adenine nucleotide translocase activity by Pi demonstrates that calcium is likely only partially involved in Pi-induced depression in oxidative phosphorylation and that a further partial contribution arises from a decrease in adenine nucleotide translocase activity. PMID- 2557035 TI - Inhibition of human neutrophil aggregation by albumin. Relationship with cytoskeleton reorganization. AB - Albumin, at concentration normally present in plasma (approximately 600 microM), significantly inhibited leukotriene B4 formation induced by a receptor mediated (fMet-Leu-Phe) and a receptor independent (calcium ionophore A23187) stimuli in human neutrophils. The inhibition of leukotriene B4 synthesis was accompanied by a concomitant reduction of neutrophil aggregation. In addition, this plasma protein prevented the increase in F-actin content of neutrophils stimulated with fMet-Leu-Phe and A23187, thus suppressing actin polymerization. These data indicate that albumin profoundly affects biochemical and functional aspects of neutrophils suggesting, for this plasma protein, a regulatory role in the overall pattern of the inflammatory reaction. PMID- 2557036 TI - Phenytoin dephosphorylates the alpha(-) catalytic subunit of (Na+, K+)-ATPase. A study in mouse, cat and human brain. AB - Phenytoin, a potent antiepileptic drug, has been thought to stimulate Na+, K+ transport across cell membranes, but its influence on (Na+, K+)-ATPase activity remains highly controversial. We have investigated the effects of the drug on the phosphorylation level of (Na+, K+)-ATPase partially purified from mouse, cat and human brain. (Na+, K+)-ATPase catalytic subunits [alpha(+) and alpha(-)] were resolved by sodium dodecylsulfate polyacrylamide gel electrophoresis. Previous experiments had shown that phenytoin dephosphorylates the (Na+, K+)-ATPase catalytic subunit by +/- 50% in C57/BL mice. In the present study, we showed that phenytoin (10(-4) M) decreases the phosphorylation level of (Na+, K+)-ATPase catalytic subunit by the same value in cat and human cortex. Moreover, that effect is predominant on the alpha(-) subunit, thought to be the predominant enzymatic form in non-neuronal or glial cells. The results are thus favoring the hypothesis that phenytoin stimulates the brain (Na+, K+)-ATPase. They further suggest that phenytoin mainly activates the glial enzymatic form, providing central nervous system with an enhanced ability to regulate extracellular K+. PMID- 2557037 TI - Inhibition of cellular activities by triethyllead. Role of glutathione and accumulation of triethyllead in vitro. AB - We investigated the interaction of triethyllead with ATP-coupled cellular enzymatic activities and the role of GSH to reverse the observed inhibition of these enzymes. Triethyllead inhibited the membrane bound Na+-K+-ATPase from HeLa cells (IC50 12 microM) and the ATP-hydrolysing activity of the mitochondrial F0 F1-ATPase complex (IC50 17 microM). Addition of 1 mM GSH reversed both enzyme activities totally, whereas lower GSH concentrations showed a less pronounced effect. Surprisingly, in freshly isolated rat liver mitochondria the ATP synthesizing activity was also inhibited by triethyllead (IC50 16 microM), in spite of a measured high intramitochondrial GSH concentration (up to 10 mM). Further experiments in isolated submitochondrial particles revealed that ATP synthesis and ATP-hydrolysis were inhibited by triethyllead with similar IC50 values, and both activities could be protected in vitro from the organolead compound in the presence of 1 mM GSH. Thus in all activities tested in vitro a high excess of GSH over triethyllead (greater than or equal to 25-fold) is necessary to restore the inhibited enzymes. The intramitochondrial triethyllead concentration was further determined after incubation of intact mitochondria with 10 microM of the organolead compound. The organolead concentration measured was as high as 600 microM. This means that in intact mitochondria there exists only a ca. 16-fold excess of GSH, which has been shown to be insufficient to protect ATP synthesizing and ATP-hydrolyzing activities of the F0-F1-ATPase from triethyllead in vitro. We concluded that in intact mitochondria the F0-F1-ATPase complex is inhibited by triethyllead due to its accumulation in the matrix. PMID- 2557038 TI - Hydroxylation of salicylate by activated neutrophils. AB - Salicylates are metabolized in vivo to hydroxylated compounds, including 2,3 dihydroxybenzoic acid and 2,5-dihydroxybenzoic acid (gentisic acid). The present study hypothesized that activated neutrophils represent one pathway for salicylate hydroxylation. Human neutrophils were incubated in medium containing 10 mM salicylate and stimulated with phorbol myristate acetate (PMA) for 1 hr. The cell-free supernatant fractions were analyzed by HPLC. Neutrophils (1 x 10(6) cells) produced 55 +/- 11 ng of gentisic acid. Neutrophils also produced smaller quantities of 2,3-dihydroxybenzoic acid. Antioxidant inhibitor experiments indicated that superoxide dismutase (SOD), heme protein inhibitors, and glutathione blocked gentisic acid formation, whereas catalase, mannitol, and deferoxamine failed to inhibit. Experiments with the reagent hypochlorous acid (HOCl) and the model myeloperoxidase (MPO) enzyme system did not support a role for the MPO pathway in gentisic acid formation. These findings demonstrate that activated neutrophils can hydroxylate salicylate by an unknown pathway. This pathway may contribute to the increased recovery of hydroxylated salicylates in patients with inflammatory disorders. PMID- 2557039 TI - Induction of DNA-protein crosslinks by antitumor 1-nitro-9-aminoacridines in L1210 leukemia cells. AB - Ledakrin [1-nitro-9-(3'-dimethylamino-N-propylamino)acridine], an antitumor drug of the 1-nitro-9-aminoacridine family, was able to induce DNA-protein crosslinks in intact L1210 leukemia cells, as demonstrated by the potassium-dodecyl sulfate precipitation technique. Ledakrin-induced DNA-protein crosslinks were not readily reversible nor were they accompanied by DNA double-strand breaks. Also, ledakrin produced virtually no crosslinks in isolated nuclei. Ledakrin-induced DNA-protein crosslinks seemed not to be mediated by topoisomerase II, unlike well-established effects of a chemically related antitumor drug, 4'-(9 acridinylamino)methanesulfon-m-anisidide (m-AMSA). Four ledakrin analogs of divergent cytotoxic potencies also induced DNA-protein crosslinks but not DNA double-strand breaks in intact L1210 cells. A significant positive correlation existed between the ability of ledakrin and its 1-nitro analogs to induced DNA protein crosslinks and the antiproliferative effects of these drugs. The results are consistent with the previously shown ability of 1-nitro-9-aminoacridines to covalently bind to macromolecules after metabolic activation in the cell. In addition to previously demonstrated DNA interstrand crosslinks and monofunctional adducts, DNA-protein crosslinks constitute another type of DNA lesion induced by 1-nitro-9-aminoacridines. PMID- 2557040 TI - Eicosanoid production and cell accumulation induced by intrapleural injection of sodium arachidonate in the rat. Characterization of the model. PMID- 2557041 TI - Inhibition by REV-5901 of leukotriene release from guinea-pig and human lung tissue in vitro. AB - The 5-lipoxygenase inhibitor REV-5901 [alpha-pentyl-3-(2 quinolinylmethoxy)benzene-methanol] was evaluated for effects on mediator release in vitro from fragmented guinea-pig and human lung. In guinea-pig lung, REV-5901 inhibited the antigen-induced release of immunoreactive leukotriene D4 (iLTD4) with an IC50 of 9.6 +/- 2.9 microM and immunoreactive leukotriene B4 (iLTB4) with an IC50 of 13.5 +/- 2.2 microM. REV-5901 also inhibited the calcium ionophore induced release of immunoreactive leukotrienes from human lung in vitro with IC50 values of 11.7 +/- 2.2 MicroM versus peptide leukotrienes and 10.0 +/- 1.1 microM versus iLTB4. The inhibition of release of iLTD4 and iLTB4 with similar IC50 values suggests that REV-5901 acts by inhibiting 5-lipoxygenase in this system. At concentrations as high as 50 microM, REV-5901 did not inhibit the release of thromboxane B2 (TxB2), 6-keto-prostaglandin-F1 alpha (6-keto-PGF1 alpha), or histamine from either lung. The lack of effect on TxB2 and 6-keto-PGF1 alpha indicates that REV-5901 did not inhibit phospholipase A2, cyclooxygenase, or thromboxane synthetase. Inhibition of leukotriene release by REV-5901 could not be reversed by washing. Among various 5-lipoxygenase inhibitors, the order of potency for inhibition of iLTD4 release from guinea-pig lung was AA-861 greater than REV-5901 greater than phenidone greater than nafazatrom greater than NDGA greater than BW755C. These findings suggest that REV-5901 is a selective and relatively potent inhibitor of leukotriene release from lung tissue in vitro. PMID- 2557042 TI - Comparison of propranolol-binding plasma proteins in sheep with those in humans, dogs and rats. AB - Alpha 1-acid glycoprotein (AGP), a plasma protein responsible for the binding of a variety of basic lipophilic drugs including propranolol, is different from other plasma proteins in being nonprecipitable after treatment with 1.2M perchloric acid (PCA). To assess the contribution of AGP to drug disposition in sheep and three other species (rats, dogs, and humans), the binding of [3H]propranolol was measured before and after PCA precipitation. PCA precipitation reduced propranolol binding 14-fold in sheep, compared to 2- to 3 fold in the other species. This implied either that sheep AGP binds less propranolol than other species, or that the AGP in sheep is more precipitable. It was not due to inherently poor propranolol binding, as whole sheep plasma bound a higher fraction than the other species. When samples of PCA-precipitated sheep plasma were analyzed using polyacrylamide gel electrophoresis, the concentration of AGP was 10-20% that of the other species. Phenobarbital induction was used as a tool to examine the changes in the plasma protein profile. Phenobarbital induced propranolol binding and AGP along with two other proteins in sheep. One of these proteins migrated similarly to AGP deglycosylated by peptide-N glycosidase F. It is postulated that the greater precipitability of propranolol binding in sheep is due to a less glycosylated form of AGP which is not important in other species. PMID- 2557043 TI - Inhibition of xanthine oxidase by 4-hydroxy-6-mercaptopyrazolo[3,4-d]pyrimidine. AB - Compound B103U, 4-hydroxy-6-mercaptopyrazolo[3,4-d]pyrimidine, was investigated as an inhibitor of human xanthine oxidase. Studies in vitro demonstrated that it was significantly more potent than oxypurinol, 4,6-dihydroxypyrazolo[3,4 d]pyrimidine. It formed an initial complex with electron-rich (reduced) human xanthine oxidase that was tighter than the corresponding complex formed by oxypurinol. The initial complexes with each inhibitor and reduced enzyme were internally rearranged into more stable complexes with first-order rate constants of 2.5 to 3 per min. However, the half-life of the isomerized (stable) complex with B103U was three to four times longer than the half-life of the analogous complex with oxypurinol. This stability was previously noted by Massey et al. (J. Biol Chem 254: 2837-2844, 1970) with B103U and bovine xanthine oxidase. The overall Ki values accounting for the initial and isomerized complexes were 5 nM for B103U and 100 nM for oxypurinol. B103U was also more potent as an inhibitor of bovine xanthine oxidase-catalyzed generation of superoxide radicals. Studies in mice revealed that the relative in vitro potency of B103U was not sustained in vivo. Compared to the inhibition of xanthine oxidase by oxypurinol, inhibition by B103U was neither more potent nor longer lasting. This shortcoming was not caused by weaker inhibition of mouse xanthine oxidase. Instead, it was the result of poor bioavailability. Plasma levels of available B103U rapidly decreased from samples of mouse and human blood because of reversible binding to serum proteins. B103U was also susceptible to oxidation. Two equivalents of H2O2 stoichiometrically oxidized the 6-thiol substituent to a sulfinic acid. This oxidized product was three orders of magnitude weaker as an inhibitor of xanthine oxidase than was B103U. PMID- 2557044 TI - Regulation of human synovial fibroblast collagenase messenger RNA by interleukin 1. AB - Interleukin-1 (IL-1) may contribute to tissue destruction in rheumatoid arthritis, in part, by inducing messenger RNA (mRNA) that encodes interstitial collagenase. In human synovial fibroblasts in vitro, IL-1 induced collagenase mRNA accumulation 6 hours after being added to the cells. High levels of mRNA remained present for at least 48 hours after treatment. The rate of transcription of collagenase in isolated nuclei peaked after approximately 6 hours of treatment with IL-1 and declined thereafter, becoming nearly undetectable by 24 hours. The persistence of mRNA, in view of the transient peak of transcription, suggested that collagenase mRNA was stable in synovial fibroblasts. The half-life of collagenase mRNA after the synoviocytes were treated with actinomycin D was approximately 27 hours, both in the presence and in the absence of IL-1. It has been noted that induction of the expression of collagenase by phorbol esters requires fos protein synthesis and is mediated through a tetradecanoyl phorbol acetate response element in the 5'-flanking region of the gene. However, we found that cycloheximide, when added to synovial fibroblast cultures up to 6 hours after treatment with IL-1, inhibited the expression of collagenase mRNA. These results suggest that fos alone is unlikely to be sufficient for collagenase expression, and that additional factors, or alternative pathways, are involved in the induction of collagenase by IL-1. PMID- 2557046 TI - Alpha 1-adrenergic receptor binding sites in post-mortal human cerebral microvessel preparations: preservation in multi-infarct dementia and dementia of Alzheimer type. AB - Cerebral cortical microvessels were prepared from control, dementia of Alzheimer type (AD/SDAT) and multi-infarct dementia (MID) autopsy cases. The microvessel yields were approx. 200 micrograms protein/g starting material, and did not differ significantly between control, MID and AD/SDAT groups. The purity of the preparations was confirmed both by light and electron microscopy and by measurement of enrichment of the endothelial markers gamma-glutamyltranspeptidase and alkaline phosphatase. Higher microvessel alkaline phosphatase activities and higher microvessel/homogenate ratios of activities of both enzymes in the MID and the AD/SDAT samples than in the control samples were found, which may be consistent with previous findings of structural abnormalities of the cerebral endothelial cells in AD/SDAT. The levels of [3H]prazosin binding did not differ significantly between control. MID and AD/SDAT samples at any [ligand] tested (0.05, 0.1, and 0.5 nM), suggesting conservation of microvessel alpha 1 adrenoceptors in MID and AD/SDAT. PMID- 2557045 TI - Proton translocation in proteins. PMID- 2557047 TI - Marked locomotor stimulation in monoamine-depleted mice following treatment with atropine in combination with clonidine. AB - The present study demonstrates that the muscarinic antagonist atropine and the alpha-adrenergic agonist clonidine, though ineffective when administered separately, produced a pronounced locomotor stimulation in monoamine-depleted mice when combined. The atropine + clonidine-induced locomotor stimulation was counteracted by both the alpha 2-adrenoceptor antagonist idazoxan and the acetylcholinesterase inhibitor physostigmine. Thus, it is clear that simultaneous manipulations with cholinergic and adrenergic systems are as effective in restoring locomotion in monoamine-depleted mice as increasing central dopaminergic tone. This finding may have implications for the treatment of a movement disorder like Parkinson's disease. PMID- 2557048 TI - [Determination of Lignans in the fruits of Schisandra chinensis (Trucz.) Baill. and S. Sphenanthern Rehd. et Wils. using HPLC and their chromatograms]. AB - The lignans contained in the fruits of Schisandra chinensis and S. sphenanthern were determined by HPLC. Their chromatograms are different. 7 samples of S. chinensis from 6 habitats contain the same lignans, but in different quantity, while 16 samples of S. sphensnthern contain lignans different both in quality and quantity. PMID- 2557049 TI - A study to compare the effects of two dentifrices on adult dental calculus formation. AB - In a three month double-blind clinical trial, a tartar control dentifrice formulation containing soluble pyrophosphates was compared to a placebo formula. Both formulations contained sodium fluoride. Subjects on the active dentifrice showed a 38% reduction in calculus severity and a 25% increase in calculus-free sites compared to the placebo group. Despite obvious differences in inactive excipients, these values are similar to reported results from other commercially available tartar control dentifrices. No untoward side effects occurred throughout the test period. PMID- 2557050 TI - Ethanol may stimulate or inhibit (Na+ + K+)-ATPase, depending upon Na+ and K+ concentrations. AB - The influence of varying the ratios of [Na+]/[K+] on the effects of alcohol (500 mg/dl) on brain (Na+ + K+)-ATPase, using a commercial porcine enzyme preparation, showed that, generally, activity was stimulated by ethanol when [Na+] less than [K+], but inhibited when [Na+] greater than [K+] (with sum kept constant at 150 mM). In addition, when [Na+]/[K+] was 15/90 mM, representative of normal intracellular levels, ethanol (500 mg/dl) stimulated the porcine enzyme, but inhibited it when [Na+]/[K+] was 144/6 mM, representative of normal extracellular levels. Similarly, in freshly prepared enzyme from highly purified rat brain synaptic membranes, ethanol (100, 300, and 450 mg/dl) stimulated when [Na+]/[K+] was 15/88 mM (representing intracellular levels), but inhibited when [Na+]/[K+] was 142/4 mM (extracellular levels). PMID- 2557051 TI - Ethanol effects on bradykinin-stimulated phosphoinositide hydrolysis in NG 108-15 neuroblastoma-glioma cells. AB - The effect of short-and long-term ethanol exposure on bradykinin-stimulated hydrolysis of phosphatidylinositol 4.5-bisphosphate (PIP2) was investigated in neuroblastoma X glioma hybrid cells (NG 108-15). Acute exposure of 50-150 mM ethanol neither influenced the bradykinin-stimulated accumulation of [3H] inositol phosphates (IP1, IP2, IP3) nor the hydrolysis of PIP2 in cells labelled with [3H]-inositol. Furthermore, ethanol (100 mM) added in the absence of agonist did not influence these parameters. However, in cells cultivated for 4 days in 100 mM ethanol, PIP2 hydrolysis and IP1, IP2 and IP3 formation after stimulation by 10(-6)-10(-5) M bradykinin was markedly inhibited while there was no effect on the basal levels or on the levels found after stimulation with low concentrations of bradykinin. The inhibitory effect of ethanol on IP accumulation became significant after 2-3 days of ethanol. PMID- 2557052 TI - Ontogeny of mu and delta opioid receptors and of neutral endopeptidase in human spinal cord: an autoradiographic study. AB - The distributions of the neutral endopeptidase 24.11 (NEP; enkephalinase) and of mu and delta opioid receptors have been studied by autoradiography in the human spinal cord during ontogenesis, mu and delta sites were assessed by using [3H]DAGO and [3H]DTLET respectively and NEP was labelled by [3H]HACBO-Gly, a NEP inhibitor. Labelling by the three markers was found at an early stage of development of the central nervous system (14 weeks) and was mainly localized in the gray matter, with highest densities in the superficial layers of the dorsal horn. Moreover [3H]DAGO also diffusely labelled the ventral motor areas. NEP and delta binding sites were localized transiently in the fasciculus gracilis at the cervical level at a fetal age of 24 weeks, an area where no enkephalin-like immunoreactivity (ELI) has been found. Conversely no opioid binding sites or NEP were observed at a fetal age of 18 weeks in the intermediolateral region where ELI fibres and cells were detected transiently. In general, a better correlation between the distribution of NEP and that of delta opioid sites was observed. Meninges contained a very high density of [3H]HACBO-Gly sites. This labelling appeared almost simultaneously with that in the spinal cord tissue and increased with maturation. An increase in labelling by the three markers appeared slightly earlier than the clustering of ELI fibres in the substantia gelatinosa. Our data show that in the human spinal cord, structural and biochemical elements involved in enkephalinergic transmission appear almost simultaneously and early in ontogeny. PMID- 2557053 TI - Pro-opiomelanocortin-derived peptides in transected and contralateral motor nerves of the rat. AB - An immunocytochemical method was used to reveal beta-endorphin and alpha melanotropin peptides in nerve profiles at the neuromuscular junctions in the soleus and extensor digitorum longus muscles of the adult rat. After unilateral section of the sciatic nerve, the proportion of endplates with immunoreactive nerve profiles increased in the denervated muscles even after the axonal material in the distal nerve segments had degenerated. When the nerve was sectioned in the mid-thigh region the proportion of immunoreactive endplates increased with time up to 24-36 h after the nerve section and then eventually declined. The incidence of immunoreactive nerve profiles also increased in the proximal stump. After unilateral sciatic nerve section there was also an increase in the incidence of immunoreactive endplates in the contralateral muscles. The increases in the contralateral muscles could be detected at an earlier time if the nerve was sectioned closer to the spinal cord. The possibility is discussed that the expression of pro-opiomelanocortin peptides in the motor nerves is tonically suppressed in the normal adult but the inhibition is released after section of the nerve. The findings indicate that a signal is transferred from the site of injury in the transected nerves to the motor nerves of the contralateral limb via a transneuronal mechanism through the spinal cord. PMID- 2557054 TI - Reduced neutrophil production of leukotriene B4 associated with AIDS. AB - Neutrophils from 10 homosexual men with evidence of HIV infection and 10 healthy controls were tested for their capacity to generate leukotriene B4. Neutrophils from patients with AIDS produced less leukotriene immunoreactivity when appropriately stimulated than neutrophils from healthy controls, whereas no significant difference could be detected between HIV-antibody-positive individuals without AIDS and healthy controls. This observation may be pertinent to the recurrence of some of the opportunistic infections associated with AIDS but more importantly, if reflecting a general defect in leukotriene production, it may provide further understanding of the mechanism which leads to reduced natural killer-cell activity, interleukin-2 and interferon-gamma production in AIDS. PMID- 2557055 TI - Statistics from the World Health Organization and the Centers for Disease Control. PMID- 2557056 TI - Lewy bodies in Alzheimer disease--one or two diseases? AB - To clarify the significance of the Lewy body (LB) in Alzheimer disease (AD), we determined the incidence and distribution of LB in 150 cases of AD and 75 controls. We also examined the pathological changes in the substantia nigra and quantified neocortical alterations, including the density of neuritic plaques, neurofibrillary tangles, and LB. LBs were present in 25% of AD cases, but in only 5% of controls. The presence of LB was associated with significantly lower numbers of neurofibrillary tangles as compared with cases of AD without LB, whereas the mean density of neuritic plaques remained unchanged. Two possibilities are discussed to explain these findings. First, the high frequency of LB in AD may reflect a predisposition of AD patients to develop idiopathic Parkinson disease. Second, the LB may represent a nonspecific cytoskeletal change in selected vulnerable neuronal populations in some subjects with AD, rather than the presence of a second pathological process. PMID- 2557057 TI - Dynein structure and function. PMID- 2557058 TI - Communication between mitochondria and the nucleus in regulation of cytochrome genes in the yeast Saccharomyces cerevisiae. PMID- 2557059 TI - Initiation of eukaryotic DNA replication in vitro. PMID- 2557060 TI - Drosophila extracellular matrix. PMID- 2557061 TI - Membrane traffic in endocytosis: insights from cell-free assays. PMID- 2557062 TI - The biogenesis of lysosomes. PMID- 2557063 TI - The role of beta receptors in the peripheral vasculature of calves with a total artificial heart. AB - Drugs given to a total artificial heart (TAH) calf isolate their vascular effects independent of the myocardium. During experiments, the TAH maintains full ejection, constant heart rate, and percent systole, and uses no vacuum. Cardiac output (CO) varies solely and directly with preload. Six calves received an infusion of isoproterenol, a beta agonist, and three calves received propranolol, a beta antagonist. The isoproterenol was resumed after beta blockade. Isoproterenol alone caused a significant increase in CO, an effect that was attenuated but not eliminated with beta blockade. Both isoproterenol and propranolol decreased AoP, but only isoproterenol increased preload. Beta receptors play a significant role in decreasing venous capacitance with increased preload and CO, independent of the myocardium. PMID- 2557064 TI - Current perception threshold. Reproducibility and comparison with nerve conduction in evaluation of carpal tunnel syndrome. AB - Neuro-selective current perception threshold (CPT) values quantify peripheral nerve (n) integrity and provide an index of adequate hemodialysis (HD). Evaluation of polyneuropathy (PN) by CPT correlates with nerve conduction testing (NCT). CPT is convenient, painless, and may be performed during HD. Early detection of carpal tunnel syndrome (CTS), a complication of uremia, permits curative intervention. Utility of CPT and NCT measurements in detecting CTS in 29 stable HD patients were evaluated. Reproducibility of seven CPT determinations over 4 weeks was determined in each of 9 HD patients. The coefficient of variation for repeated 2000 Hz CPT measures was 6%. PN was detected by CPT in 92% of the patients and by NCT in 79% (r = 0.79, p less than 0.001). In 38% of the hands there was a CPT impairment in both the median and ulnar nerves (n), of which 25% were symptomatic for CTS. CPTs consistent with CTS (sufficiently greater impairment of the median vs ulnar n) were observed in 31% of the hands with combined median and ulnar n CPT abnormalities, and 11% were identified with CTS by NCT. The unique ability of the CPT exam to quantify hyperesthesia may account for its superior CTS detection sensitivity. These findings demonstrate that repeated CPT determinations are consistent and are diagnostic for CTS. PMID- 2557065 TI - A new improved biodegradable tracheal prosthesis using hydroxy apatite and carbon fiber. AB - A new biodegradable tracheal prosthesis was developed using hydroxyapatite rings as the artificial tracheal cartilage, a carbon fiber tube as the tracheal tube; it was then implanted into the cervical trachea in dogs. Morphologic examination revealed that the hydroxyapatite ring was anchored firmly to the tracheal cartilage by ingrowth of cartilaginous tissue into the macropores of the hydroxyapatite. PMID- 2557066 TI - Development of a novel biocompatibility test method based on intracellular calcium ion mobilization. AB - This paper presents a novel evaluation method for determining platelet/material interaction at the transmembrane level by incorporation of a Ca-sensitive fluorophore (Fura-2) into platelets. The transmembrane stimulation of platelets which are eluted from commercially available artificial dialyzers was quantitatively measured as the elevation of intracellular free calcium ion concentration. This method clearly differentiated platelet response to hydrophilic (ethylene vinyl alcohol copolymer and cellulose) vs. hydrophobic (polyacrylonitrile and cellulose acetate) membranes, and offers a versatile means to understanding platelet/material interaction. PMID- 2557067 TI - Novel mechanical assistance in the treatment of endotoxic and septicemic shock. AB - Systemic sepsis is a frequent and fatal complication of postoperative patients. More recently, therapeutic trials of plasma or blood exchange are performed in septic patients for the purpose of reducing both endotoxins and albumin-bound toxins. As an alternate approach such as this for the removal of endotoxin directly from the blood, the authors recently developed polymyxin B immobilized fiber (PMX-F) as a biomaterial for selectively detoxifying endotoxin. That this newly invented PMX-F neutralizes a sufficient amount of endotoxin in vitro was reported previously. In ex vivo experiments, direct hemoperfusion by PMX-F was performed on purified endotoxin injected canine and on live Escherichia coli induced septic dogs. Only 5% (1/20) survived in the control group, but 75% (30/40) survived in the treated group. Septic dogs died within 18 hours after bacteria infusion in the control group. But all in the treated group survived 3 days. Forty percent of them survived permanently. These observations indicate that PMX-F treatment, namely selective removal of endotoxin in the endotoxic and septicaemic dogs prolongs or increases the chances of survival. Blood compatibility of PMX-F was also evaluated both in vitro and in vivo. With platelets it was shown to be fairly good and with red blood cells, white blood cells, and proteins, extremely good. A preclinical study on the efficacy and safety of PMX-F throughout widespread experiments has just ended. PMID- 2557068 TI - The role of the protein C-thrombomodulin system in physiologic anticoagulation during cardiopulmonary bypass. AB - Changes in the protein C-thrombomodulin (PC-TM) system in relation to other coagulofibrinolytic parameters were examined in 25 patients undergoing open heart surgery. Although all patients were given heparin, a decrease in antithrombin III (ATIII) and progressive increase in thrombin-ATIII complex (TAT) and fibrinopeptide A (FPA) levels were noted during cardiopulmonary bypass, which indicated that heparinization did not completely inhibit the formation of thrombin and its function. C1-inactivator (INA) resistant fibrinolytic activity increased markedly during CPB, in parallel with the change of tissue plasminogen activator antigen (t-PA;Ag), which indicates that fibrinolytic activity during CPB is mainly of extrinsic origin caused by t-PA. Protein C antigen (PC;Ag), protein S antigen (PS;Ag), and thrombomodulin antigen (TM;Ag) were all decreased significantly during CPB. This is considered to reflect the activation and consumption of the PC-TM system in response to generated thrombin. Furthermore, enhancement of the extrinsic fibrinolytic system is easily explained by the action of activated PC, which counteracts plasminogen activator inhibitor (PAI 1), to cause enhancement of t-PA activity. PMID- 2557069 TI - Development of a compliant surgical adhesive derived from novel fluorinated hexamethylene diisocyanate. AB - We have recently developed a novel compliant surgical adhesive, based on urethane chemistry, that imparts excellent in vivo performance, except for a potential carcinogenic problem associated with aromatic diisocyanates. In this article, a chronic histopathologic study was conducted by implantation of adhesives in subcutaneous and hepatic tissues for up to 20 months. Neither carcinogenic episodes nor tissue abnormalities were found. Based on the structure-carcinogenic potential relationship of diisocyanates, fluorinated hexamethylene diisocyanate (HMDI) was found to be noncarcinogenic, despite its high nucleophilic reactivity. The fluorinated HMDI based adhesive was defined as the most reliable adhesive, with best in vivo performance, including ensured nontoxicity. PMID- 2557070 TI - A new method of determining the solute permeability of hollow-fiber dialysis membranes by means of laser lights traveling along optic fibers. AB - To develop a new method of determining solute permeability more simply and accurately, the authors employed light from a laser traveling along quartz optic fibers. Dialysis experiments at 310 K were made with a single hollow fiber containing aqueous test solutes. A membrane tube was sealed at either end with quartz optic fibers. Helium-neon and helium-cadmium laser lights emitted from one of these optic fibers into the test solution at wavelengths of 543 and 442 nm for vitamin B12 and cytochrome-C, respectively, were caught by the other optic fiber and detected with a silicon photodiode. The solute permeability for cytochrome-C obtained by this method was almost in agreement with that for beta-2 microglobulin by the radioisotope method. This study demonstrates the usefulness of light from a laser traveling along quartz optic fibers in determining the solute permeability of hollow-fiber dialysis membranes. PMID- 2557071 TI - Use of ganciclovir in the treatment of cytomegalovirus infections. AB - Ganciclovir (Cymevene*) is an antiviral drug that shows efficacy against cytomegalovirus (CMV) infections. These are particularly important in patients with AIDS or iatrogenic immunosuppression. Ganciclovir is a virostatic agent that prevents viral DNA replication: it is effective in CMV retinitis, colitis and pneumonitis. Since the drug is particularly taken up by cells with a high rate of DNA replication, its adverse effects are most pronounced in bone marrow and testis; its present indications are therefore only for serious CMV disease. PMID- 2557072 TI - Novel variants of human papillomavirus type 2 in warts from immunocompromised individuals. AB - Twenty cutaneous warts from I8 renal transplant recipients (RTR) and a single wart sample from a patient suffering from Hodgkin's disease were screened for the presence of human papillomavirus (HPV) sequences by Southern blot analysis. Thirteen samples were shown to contain HPV DNA sequences, one was identified as HPV-3, two as HPV-4, six as HPV-2 and four were unidentifiable by restriction digest analysis. HPV-3 was identified in the wart sample from the patient suffering from Hodgkin's disease. In the six samples in which HPV-2 was found, the viral genome showed polymorphism for the PvuII and PstI sites. Two PvuII patterns and one PstI pattern are novel. PMID- 2557073 TI - Intraosseous adenoid cystic carcinoma of the mandible. AB - Radiolucent lesions in the jaws usually are associated with pathology of odontogenic origin. An unusual case of an adenoid cystic carcinoma arising from within the mandible is described. The origin of central salivary tumours is discussed and the literature reviewed. PMID- 2557074 TI - Human adipose tissue hormone-sensitive lipase: identification and comparison with other species. AB - The mRNA for human hormone-sensitive lipase (HSL) was identified using Northern blot analysis and a cDNA-probe for rat HSL. As in the rat, human adipose tissue expresses a single mRNA species of 3.3 kb. Using Western blotting with a polyclonal rabbit antibody towards rat adipose tissue HSL, the corresponding enzyme in human adipose tissue was identified with an apparent 88 kDa polypeptide, thus slightly larger than the rat and bovine 84 kDa, and the mouse and guinea-pig 82 kDa species. Additional evidence for the identification was provided by the inhibition of HSL diacylglycerol lipase activity by the anti-rat HSL antibody, and by NaF, DFP and Hg2+, known inhibitors of HSL. The concentration of the enzyme, as reflected by its activity per g tissue and the specific activity was about two thirds of that in the rat adipose tissue (200 g rats). The identification of the human enzyme protein made it possible to directly demonstrate its phosphorylation by cAMP-dependent protein kinase, thus extending the previous report regarding activation of the lipase with this kinase and ATP-Mg2+ in human adipose tissue extracts (Khoo, J.C., Aquino, A.A. and Steinberg, D. (1974) J. Clin. Invest. 53, 1124-1131). PMID- 2557076 TI - Characterization of cytosolic forms of CTP: choline-phosphate cytidylyltransferase in lung, isolated alveolar type II cells, A549 cell and Hep G2 cells. AB - The subcellular forms of cytidylyltransferase (EC 2.7.7.15) in rat lung, rat liver, Hep G2 cells, A549 cells and alveolar Type II cells from adult rats were separated by glycerol density centrifugation. Cytosol prepared from lung, Hep G2 cells, A549 cells and alveolar Type II cells contained two forms of the enzyme. These species were identical to the L-Form and H-Form isolated previously from lung cytosol by gel filtration. Liver cytosol contained only the L-Form. Rapid treatment of Hep G2 cells with digitonin released all of the cytoplasmic cytidylyltransferase activity. The released activity was present in both H-Form and L-Form. The molecular weight of L-Form was determined from sedimentation coefficients and Stokes radius values to be 97,690 +/- 10,175. Thus, the L-Form appears to be a dimer of the Mr 45,000 catalytic subunit. The f/f degrees value of 1.5 indicated that the protein molecule has an axial ratio of 10, assuming a prolate ellipsoid shape. The estimated molecular weight of the H-Form was 284,000 +/- 25,000. The H-Form was dissociated into L-Form by incubation of cytosol at 37 degrees C. Triton X-100 (0.1%) and chlorpromazine (1.0 mM) also dissociated the H Form into L-Form. Western blot analysis indicated that both forms contained the catalytic subunit. An increase in Mr 45,000 subunit coincided with the increase in cytidylyltransferase activity in L-Form, which resulted from the dissociated of H-Form. The L-Form was dependent on phospholipid for activity. The H-Form was active without lipid. Phosphatidylinositol was present in the H-Form isolated from Hep G2 cells. The phosphatidylinositol dispersed when the H-Form was dissociated into L-Form. Phosphatidylinositol and phosphatidylglycerol cause L Form to aggregate into a form similar to H-Form. Phosphatidylcholine/oleic acid (1:1 molar ratio) and oleic acid also aggregated the L-Form. Phosphatidylcholine did not produce aggregation. We conclude that the H-Form is the active form of cytidylyltransferase in cytoplasm. The H-Form appears to be a lipoprotein consisting of an apoprotein (L-Form dimer of the Mr 45,000 subunit) complexed with lipids. A change in the relative distribution of H-Form and L-Form in cytosol would alter the cellular activity and thus may be important in the regulation of phosphatidylcholine synthesis. PMID- 2557075 TI - Oligomers of prostaglandin B1 inhibit arachidonic acid mobilization in human neutrophils and endothelial cells. AB - The previous paper (Biochim. Biophys. Acta 1006 (1989) 272-277) has demonstrated that oligomers of prostaglandin B1 are effective in vitro inhibitors of a wide range of both cell-derived and extracellular phospholipases A2. The present study has investigated the effects of prostaglandin oligomers on agonist-stimulated phospholipase activity on intact human cells. PGBx, an oligomer (n = 6) or PGB1, and PGB-trimer inhibit as much as 95% of the A23187-stimulated release of arachidonic acid from human neutrophils. The effect is dose-dependent, with an IC50 of 4-5 microM; near maximal inhibition is obtained with as little as 1 min of preincubation with PGB-trimer. Consistent with its role as a phospholipase A2 inhibitor, PGB-trimer also inhibits the A23187-stimulated incorporation of [3H]acetate into platelet-activating factor. PGBx and PGB-trimer also inhibit the release of arachidonic acid from human umbilical vein endothelial cells stimulated with histamine, thrombin, or ionophore A23187; inhibition of the basal or unstimulated turnover of both arachidonic acid and oleic acid is also observed. Inhibition by PGB-trimer can be blocked by simultaneous addition of 50 microM albumin; cells preincubated with PGB-trimer are not affected by albumin. Furthermore, removal of exogenous PGB-trimer prior to challenge with A23187 does not reverse the inhibition of either endothelial cells and neutrophils. Thus, prostaglandin B1 oligomers are taken up by human neutrophils and vascular endothelial cells and serve as potent inhibitors of arachidonic acid mobilization. One mechanism for the pharmacological effects of PGBx may be inhibition of cell-associated and extracellular phospholipase A2. PMID- 2557077 TI - Effects of the wasp venom peptide, mastoparan, on a phosphoinositide-specific phospholipase C purified from rabbit brain membranes. AB - The wasp venom peptide, mastoparan (Ile-Asn-Leu-Lys-Ala-Leu-Ala-Ala-Leu-Ala-Lys Lys-Ile-LeuNH2), activated phosphatidylinositol 4,5-bisphosphate (PIP2) hydrolysis as catalyzed by a phosphoinositide-specific phospholipase C (PLC-Im) purified from rabbit brain membranes. This activation was found when the molar ratio of mastoparan to PIP2 was less than 1 and when the concentration of PIP2 exceeded 10 microM. PIP2 breakdown was inhibited at both high and low substrate concentrations if the molar ratio of mastoparan to PIP2 was greater than 1. The stimulatory effect of mastoparan correlated with its ability to restrict aggregation of PIP2 into higher order structures (liposomes or mixed deoxycholate/phospholipid micelles) as the concentration of PIP2 was increased to 10 microM or greater. Mastoparan stimulation of PIP2 breakdown required the presence of a higher calcium concentration than was necessary for detection of enzyme activity. Both the stimulatory and inhibitory effects of mastoparan on PIP2 hydrolysis were lost if 2.5 mM deoxycholate was present in the assays. Hydrolysis of phosphatidylinositol (PI) by PLC-Im was inhibited at all concentrations of mastoparan tested. These results show that both PIP2 and PI are suitable substrates for PLC-Im, depending on the physical characteristics of their aggregates in aqueous suspension. An amphiphilic alpha-helix-forming peptide such as mastoparan may modulate phospholipase C activity due to the peptide's interaction with phospholipid substrates. PMID- 2557078 TI - On the presence of the chromosomal proteins HMG I and HMG Y in rat organs. AB - Using antiserum raised against HMG I, we have shown that HMG I and HMG Y are present in perchloric acid extracts of kidney, lung, heart, brain, liver and intestine in the rat, suggesting that the expression of these proteins may not be dependent upon proliferative activity. The results also show that the ratio between HMG I and HMG Y varies between different organs. PMID- 2557079 TI - Isolation of a cDNA likely to encode a novel Ca2+-dependent/calmodulin-stimulated protein phosphatase. AB - Complementary DNA encoding a novel protein phosphatase catalytic subunit has been isolated from a rabbit brain library. The deduced protein sequence is more similar to the major Ca2+-dependent/calmodulin-stimulated protein phosphatase (2B) in brain (55% identity) than to protein phosphatases 1 and 2A (38-39% identity). A putative calmodulin-binding domain is present C-terminal to the catalytic domain, which closely resembles that of the mouse brain enzyme. These findings represent the first indication that at least two distinct Ca2+ dependent/calmodulin-stimulated protein phosphatases are present in mammalian brain. PMID- 2557080 TI - Antibody targeted liposomes containing poly(rI).poly(rC) exert a specific antiviral and toxic effect on cells primed with interferons alpha/beta or gamma. AB - Double-stranded RNA can stimulate interferon production and mediate an antiproliferative effect on certain cell types. We evaluated the possibility of specifically targeting to cells in vitro the RNA duplex poly(rI).poly(rC) in pharmacologically active form after its encapsulation in small, unilamellar liposomes, to which was covalently coupled protein A. These liposomes became bound to and were endocytosed by murine L929 cells in the presence of protein A binding monoclonal antibodies specific for an expressed cell surface protein, the H-2K molecule. When L929 cells were preincubated in the presence of low doses of interferon alpha/beta or gamma, they could be activated to produce interferon following exposure to either free poly(rI).poly(rC), or specifically bound liposomes poly(rI).poly(rC), but not the same liposomes in the presence of non cell binding control antibodies. Specifically bound liposome-encapsulated poly(rI).poly(rC) was toxic to L929 cells at dose levels at least three logs lower than free poly(rI).poly(rC). This toxicity was also dependent on pre treatment with interferon. These results indicate that liposome-encapsulated poly(rI).poly(rC) can survive endocytosis and can be released in active form to specific cell populations, at concentrations much lower than that required for pharmacologic effects of the same molecule in free form. They suggest that introduction into cells of other nucleic acids might benefit from the antibody targeted liposome technology described here. PMID- 2557081 TI - Transepithelial electrical responses to Cl- of nonsensory region of gerbil utricle. AB - Sheets of utricular epithelium from gerbil were mounted in a micro-Ussing chamber in order to identify and localize chloride conductances. The [Cl-] was rapidly reduced (substituted with isethionate) in the apical or basolateral perfusate and the transepithelial potential difference (Vt) and transepithelial resistance (Rt) were monitored continuously. In addition, agents known to inhibit anion transport in other epithelia were applied. The direction of all initial changes in Vt and Rt due to Cl- substitutions were consistent with the presence of ionic conductances for Cl- on both sides of the epithelium. The time-courses and magnitudes of the fall in Vt and increase in Rt during apical [Cl-] steps in the presence and absence of basolateral bumetanide were monophasic and identical in the two cases. The response of Vt to basolateral [Cl-] steps was biphasic and the initial response was greatly attenuated by bumetanide. These findings demonstrate that the largest conductance for Cl- is in the basolateral cell membrane, but that the paracellular and/or apical pathway also possess a finite Cl- conductance. All three agents tested, 3',5-dichlorodiphenylamine-2-carboxylic acid (DCDPC), 5-nitro-2(3-phenylpropylamino)benzoic acid (NPPB) and 4,4' diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), caused an increase in Vt. NPPB and DIDS were more effective from the apical side. DCDPC and DIDS administered from the apical side led to a decrease in Rt. These results suggest that these agents act in this tissue by enhancing a conductive pathway on the apical membrane rather than blocking the basolateral Cl- conductance. PMID- 2557082 TI - Opioid receptors in rat cardiac sarcolemma: effect of phenylephrine and isoproterenol. AB - The present study demonstrates the presence of opioid receptors in the rat cardiac sarcolemma isolated by the hypotonic LiBr-shock procedure. Opioid binding was measured by using [3H]U69 593, [3H](2-D-penicillamine,5-D penicillamine)enkephalin ([3H]DPDPE) or [3H][D-Ala2,MePhe4,Gly-(ol)5]enkephalin ([3H]DAGO) as selective radioligands for K, delta and mu opioid receptors, respectively. Both the K- and delta-selective ligands exhibited highly specific (75-86%) binding, saturable at a concentration of about 20 nM. No specific binding for the selective agonist DAGO was observed. A marked increase in both [3H]U69 593 and [3H]DPDPE binding was observed after incubation of the sarcolemma with the alpha-adrenoceptor agonist phenylephrine or with the beta-adrenoceptor agonist isoproterenol. These stimulatory effects were associated with an increase in the Bmax values, a decrease in the Kd values, and were completely antagonized by the respective antagonists phentolamine and propranolol. PMID- 2557083 TI - Sidedness of the effect of amines on the steady-state phosphorylation level of reconstituted Na+/K+-ATPase. AB - The sidedness of the effects of several amines on the steady-state phosphorylation level of rabbit kidney Na+/K+-ATPase has been studied with the enzyme incorporated in phosphatidylcholine-cholesterol containing proteoliposomes. The presence of ouabain prevented phosphorylation of non incorporated or rightside-out incorporated enzyme, so that only the inside-out incorporated Na+/K+-ATPase molecules were studied. Addition of either Na+ or several amines to the extracellular side of the enzyme led to an enhancement of the steady-state phosphorylation level obtained with optimal concentrations of Na+, Mg2+ and ATP at the cytosolic side. The series imidazole greater than Na+ greater than triallylamine greater than Tris greater than ethylenediamine showed a decrease in affinity. Histidine, sorbitol and choline chloride had no effect at the extracellular side. This means that in addition to the well-known cytosolic ligands either Na+ or a positively charged amine buffer has to be present extracellularly in order to obtain an optimal phosphorylation level. At the cytoplasmic side the tested amines exerted different effects. (i) Imidazole and triallylamine enhanced the steady-state phosphorylation level when the extracellular conditions were optimal (saturating amine concentration). (ii) Tris and ethylenediamine decreased the steady-state phosphorylation level and (iii) histidine had no effect. The cytoplasmic effects of the amine compounds correlate with those described by Schuurmans Stekhoven et al. (Biochim. Biophys. Acta 937 (1988) 161-171) for the unsided preparation. The extracellular effects, however, are apparently masked in experiments with fragmented enzyme preparations and are assumed to be potentiating effects which make the enzyme ready for phosphorylation upon a cytoplasmic trigger (e.g. Na+). PMID- 2557084 TI - The lateral organization of components of the membrane skeleton and superoxide generation in the plasma membrane of stimulated human neutrophils. AB - Studies were performed to examine the lateral organization of the NADPH oxidase system in the plasma membrane of human neutrophils. Analysis of the subcellular fractionation of human neutrophils by isopycnic sedimentation of cavitated cell lysates suggested that there may be more than one population of plasma membrane vesicles formed upon cell disruption. One population (30-32% sucrose) contained surface accessible wheat germ agglutinin binding sites, alkaline phosphatase activity, and cytochrome b. Another population (34-36% sucrose) contained membrane-bound flavin and, when the cells were prestimulated with phorbol myristate acetate (PMA), NADPH-dependent superoxide generating activity. Approximately 25% of the neutrophil cytochrome b cosedimented with the heavy population, confirming our previous hypothesis (Parkos et al. (1985) J. Biol. Chem. 260, 6541-6547) that only a fraction of the total cellular cytochrome b is involved in superoxide production. The heavy plasma membrane fraction was also enriched in membrane associated actin and fodrin as detected by Western blot analysis. After extraction of the plasma membrane vesicles with detergent cocktails, the majority of superoxide generating activity remained associated with the detergent insoluble pellet. Western blot analysis demonstrated that the pellets were also enriched in actin. Further analysis of these pellets using rate zonal detergent-containing sucrose density gradients indicated that the superoxide generating complex had an approximate sedimentation coefficient of 80 S, suggesting that the neutrophil superoxide generating system may form a complex on the plasma membrane which is associated with or somehow organized by the membrane skeletal matrix. This organization may be of functional relevance not only to the actual production of superoxide, but also to the targeting of microbicidal oxidants. PMID- 2557086 TI - Oncogenes, growth factors and hematopoietic cell transformation. PMID- 2557085 TI - Topoisomerase-targeting antitumor drugs. AB - Much has been learned about the unusual type of DNA damage produced by the topoisomerases. The mechanism by which these lesions trigger cell death, however, remains unclear, but it appears that DNA metabolic machinery transforms reversible single-strand cleavable complexes to overt strand breaks which may be an initial event in the cytotoxic pathway. For the topoisomerase I poisons, they produce breaks at replication forks that appear to be the equivalent of a break in duplex DNA. Indicating that this may be an important cytotoxic lesion is the hypersensitivity to camptothecin of the yeast mutant rad52, which is deficient in double-strand-break-repair. The topoisomerase poisons preferentially kill proliferating cells. In the case of the topoisomerase I poison camptothecin, dramatic S-phase-specific cytotoxicity can explain its preferential action on proliferating cells. For the topoisomerase II poisons, high levels of the enzyme in proliferating cells, and very low levels in quiescent cells appear to explain the resistance of quiescent cells to the drug's cytotoxic effects. Thus, the topoisomerase poisons convert essential enzymes into intracellular, proliferating cell toxins. The identification of both topoisomerase I and II as the specific targets of cancer chemotherapeutic drugs now provides a rational basis for the development of topoisomerase I poisons for possible clinical use. Knowledge of the molecular mechanisms of cell killing may lead to the identification of new therapies for treating cancer. The topoisomerase poisons appear to be a good tool for studying cell killing mechanisms as they produce highly specific and reversible lesions. PMID- 2557087 TI - Cytidylate cyclase activity in mouse tissues: the enzymatic conversion of cytidine 5'-triphosphate to cytidine 3',5'-cyclic monophosphate (cyclic CMP). AB - Cytidylate cyclase activity, which enzymatically converts cytidine 5' triphosphate (CTP) to cytidine 3',5'-cyclic monophosphate (cyclic CMP), has been demonstrated in mouse tissue homogenates by use of a highly sensitive enzyme immunoassay (EIA) specific for cyclic CMP. Cyclic CMP formation is dependent on the amount of homogenate and on the incubation time. Although the enzyme activity was detected at wide ranges of pH from 6.8 to 11.5, the maximal activity was observed at around pH 9.4. The optimal temperature was 37 degrees C. Cytidylate cyclase activity was almost completely lost if the homogenates were heated at 90 degrees C for 3 min prior to use. The enzyme reaction exhibited typical Michaelis Menten kinetics with an apparent Km for CTP of approx. 0.31 mM. Cyclic CMP formation was greatly enhanced with 4 mM Mn2+, Mg2+, Co2+; Mn2+ was the most effective. Fe2+ and Ca2+ were without effect. Cu2+ and Zn2+ at a concentration of 0.1 to 0.5 mM were inhibitory to Mn2+-dependent activity. Moreover, the enzyme activity was inhibited by several nucleotides including ATP, ADP, 5'-AMP, and GTP. Cytidylate cyclase activity was found to be present in all homogenates from a variety of mouse tissues examined except heart, with the highest level found in brain, and the lowest in liver. PMID- 2557089 TI - Biosyntheses of interstitial collagens and fibronectin by porcine aorta smooth muscle cells. Modulation by low-molecular-weight heparin fragments. AB - The effect of low-molecular-weight heparin fragments (CY222) on the biosynthetic phenotype of porcine aortic smooth muscle cells (SMC) was investigated in vitro on overconfluent cell cultures. Addition of increasing concentrations of CY222 to the culture medium of early passage SMC resulted in a dose-dependent decrease of type III to type I collagen ratio without change in total collagen biosynthesis. In the same range of concentrations CY222 did not affect the biosynthesis of fibronectin. However, heparin fragments decreased the proportion of the freshly synthesized pericellular form of fibronectin with a concomitant increase of neosynthesized intracellular fibronectin, indicating an inhibitory effect of CY222 on fibronectin secretion. Our results demonstrate that the biosynthetic phenotype of SMC in vitro can be modulated by low-molecular-weight heparin fragments and confirm also that interactions between cells and extracellular matrix molecules can modify the biosynthetic pattern of mesenchymal cells. PMID- 2557088 TI - Amiloride activation of hepatic microsomal glucose-6-phosphatase; activation of T1? AB - The mechanism of activation of hepatic microsomal glucose-6-phosphatase (EC 3.1.3.9) in vitro by amiloride has been investigated in both intact and fully disrupted microsomes. The major effect of amiloride is a 4.5-fold reduction in the Km of glucose-6-phosphatase activity in intact diabetic rat liver microsomes. Amiloride also decreased the Km of glucose-6-phosphatase activity in intact liver microsomes isolated from starved rats 2.5-fold. Kinetic calculations, direct enzyme assays and direct transport assays all demonstrated that the site of amiloride action was T1, the hepatic microsomal glucose 6-phosphate transport protein. This is, to our knowledge, the first report of an activation of any of the proteins of the multimeric hepatic microsomal glucose-6-phosphatase complex. PMID- 2557090 TI - Collagen synthesis in explants from rat intestine. AB - Collagen is the major structural component of the intestinal wall and its metabolism is of special interest for intestinal strength. We describe collagen synthesis in short-term (3 h) incubations of rat intestinal tissue, as measured in terms of incorporation of [3H]proline in collagenase-digestible protein and percentage relative collagen synthesis. In this time span, incorporation of [3H]proline in collagen increases linearly with time and tissue weight. Addition of unlabeled proline during incubation, in excess of the 0.1 microM [3H]proline always present, strongly increases both total protein and collagen synthesis, suggesting that proline transport is rate limiting. Further experiments have been performed in the presence of labeled proline alone and with the addition of 0.35 mM unlabeled proline. Collagen synthesis is significantly higher in colon than in ileum, comprising 0.37 and 0.21%, respectively, of total protein synthesis. Also, collagen synthesis decreases considerably with age, both in ileum and colon. The results presented here demonstrate that rat intestinal explants synthesize measurable amounts of collagen in vitro and that the system used is able to detect changes in in vivo synthetic capability such as those induced by ageing. PMID- 2557091 TI - The extraction and purification of a peptide from rat insulinoma tissue. AB - A peptide was extracted and purified from rat insulinoma tissue which, although similar, was not identical to normal rat C peptides. The purity of the peptide, called rat insulinoma peptide (RIP), was investigated using polyacrylamide gel electrophoresis, isoelectric focusing and high-performance liquid chromatography. It appears to contain two peptides similar to each other but differing in their isoelectric points. The peptides as assessed by fast atom bombardment mass spectrometry have molecular masses in the region of 1982 Da, given a chain length of approx. 22 amino-acid residues. Evidence obtained using an established rat C peptides radioimmunoassay suggests that RIP shares a common C-terminus with rat C peptides. The antiserum produced to RIP was used to develop a radioimmunoassay using a tracer prepared by iodinating purified tyrosylated RIP. PMID- 2557092 TI - Interaction between catalytic and regulatory sites of the pyruvate dehydrogenase from Escherichia coli studied by the ESR technique. AB - The accessibility of sulfhydryl groups at the pyruvate dehydrogenase component of the pyruvate dehydrogenase multienzyme complex from Escherichia coli was reinvestigated. Hydrophobic interactions appear to control the reactivity of an essential cysteine residue at the active site with thiol reagents. This explains why the essential cysteine residue reacts only with thiol reagents of minor polarity, like p-hydroxymercuribenzoate or phenylmercuric nitrate, but not with Ellman's reagent or jodoacetamide. The pyruvate dehydrogenase component was modified with a nitroxide derivative of p-hydroxymercuribenzoate. The ESR spectrum of the spin-labelled enzyme changed dramatically upon addition of the cofactors thiamine diphosphate and Mg2+. Obviously spin-spin interaction occurs under these conditions caused by a transition of an inactive to an active state of the enzyme. The same conformational change is observed when the allosteric activator AMP instead of the cofactors was bound to the enzyme. The implications of these results for the allosteric regulation of the pyruvate dehydrogenase complex are discussed. PMID- 2557093 TI - Isocitrate dehydrogenase kinase/phosphatase. AB - In Escherichia coli, isocitrate dehydrogenase (IDH) is regulated by phosphorylation. This phosphorylation cycle is catalyzed by an unusual, bifunctional protein:IDH kinase/phosphatase. IDH kinase/phosphatase is expressed from a single gene, aceK, and both activities are catalyzed by the same polypeptide. The amino acid sequence of IDH kinase/phosphatase does not exhibit the characteristics which are typical of other protein kinases, although it does contain a consensus ATP binding site. The available evidence suggests that the IDH kinase and IDH phosphatase reactions occur at the same active site and that the IDH phosphatase reaction results from the back reaction of IDH kinase tightly coupled to ATP hydrolysis. The function of the IDH phosphorylation cycle is to control the flux of isocitrate through the glyoxylate bypass. This pathway is essential for growth on acetate because it prevents the quantitative loss of the acetate carbons as CO2 in the Krebs' cycle. IDH kinase/phosphatase monitors general metabolism by responding to the levels of a wide variety of metabolites, many of which activate IDH phosphatase and inhibit IDH kinase. The ability of IDH kinase/phosphatase to monitor general metabolism allows. the IDH phosphorylation cycle to compensate for substantial perturbations of the system, such as a 15 fold overproduction of IDH. The significance of the cellular level of IDH kinase/phosphatase has also been evaluated. The level of this protein is in great excess of that required for steady-state growth on acetate. In contrast, IDH kinase/phosphatase is, in some cases, rate-limiting for the dephosphorylation of IDH which results when preferred carbon sources are added to cultures growing on acetate. PMID- 2557094 TI - Studies of the phosphorylation of Escherichia coli isocitrate dehydrogenase. Recognition of the enzyme by isocitrate dehydrogenase kinase/phosphatase and effects of phosphorylation on its structure and properties. AB - Escherichia coli isocitrate dehydrogenase is completely inactivated by phosphorylation of a single serine residue per subunit. We have examined the conformations of the active and phosphorylated forms of the enzyme using circular dichroism spectroscopy. The results support the view that phosphorylation prevents the binding of NADP, probably by direct blocking of the coenzyme-binding site. Labelling studies suggest that an arginine residue at the coenzyme-binding site may be close to the phosphorylatable serine residue. The phosphorylation of isocitrate dehydrogenase is thus unusual in that it occurs at the active site of the enzyme. We therefore investigated the recognition of isocitrate dehydrogenase by isocitrate dehydrogenase kinase/phosphatase. The kinase activity of this enzyme can phosphorylate intact isocitrate dehydrogenase but not proteolytic fragments derived from it, nor a synthetic peptide corresponding to the sequence round the phosphorylation site. PMID- 2557095 TI - Metabolite-sensitive, ATP-dependent, protein kinase-catalyzed phosphorylation of HPr, a phosphocarrier protein of the phosphotransferase system in gram-positive bacteria. AB - In this review article we summarize the recent information available concerning important mechanistic and physiological aspects of the protein kinase-mediated phosphorylation of seryl residue-46 in HPr, a phosphocarrier protein of the phosphoenolpyruvate: sugar phosphotransferase system in Gram-positive bacteria. Emphasis is placed upon the information recently obtained in two laboratories through the use of site-specific mutants of the HPr protein. The results show that (i) in contrast to eukaryotic protein kinases, the HPr(ser) kinase recognizes the tertiary structure of HPr rather than a restricted part of the primary sequence of the protein; (ii) like seryl protein kinases of eukaryotes, the HPr(ser) kinase can phosphorylate a threonyl residue, but not a tyrosyl residue when such a residue replaces the regulatory seryl residue in position-46 of the protein; (iii) the regulatory consequences of seryl phosphorylation are due to the introduction of a negative charge at position-46 in the protein rather than the bulky phosphate group; and (iv) PTS protein-HPr interactions influence the conformation of HPr, thereby retarding or stimulating the rate of kinase catalyzed seryl-46 phosphorylation. The physiological consequences of HPr(ser) phosphorylation in vivo are still a matter of debate. PMID- 2557097 TI - Glycosaminoglycan conformations and changes on periodate oxidation. AB - The progressive periodate oxidation of glycosaminoglycans (GAG), including hyaluronate (HA), chondroitins (CH) (chondroitin, chondroitin 4- and 6-sulfate), dermatan sulfate (DS), and keratan sulfate (KS), were monitored by CD and high performance liquid chromatography (HPLC) using a size-exclusion column. The rate of oxidation also was measured and calculated using first- and second-order kinetics, and the data appear to fit better with first-order kinetics. In both HA and CH, the n - pi amide band at 208 nm decreases in intensity upon oxidation, but in HA it becomes positive after 16 h of periodate treatment. In CH, the band disappears, and the pi - pi amide band below 200 nm becomes optically active. Concomitantly, a second negative band near 290 nm appears for these two oxidized GAG. Oxidation causes a slight change in the CD of DS. It ordinarily displays a very weak n - pi band at 210 nm, but instead shows an intense pi - pi amide band near 190 nm. CD of KS remains unaffected by periodate. Kinetic studies, however, show a higher oxidation rate for DS than HA and CH. With the exception of KS, all other oxidized polymers shown an apparent decrease in molecular weight (higher peak retention time) in HPLC analysis. Both CD and HPLC results have been attributed to a major conformational change of HA and CH, and a minor one for DS. The ease and extent of periodate oxidation as well as the changes in molecular properties following periodate treatment are critically dependent on the configuration of the individual GAG rather than the oxidation rate. There is a distinct difference in the conformational change between HA and CH, as manifested by their dichroic behavior, that was attributed to the equatorial disposition of C-4 hydroxyl group in HA and axial disposition CH. PMID- 2557096 TI - Carbohydrate uptake in the oral pathogen Streptococcus mutans: mechanisms and regulation by protein phosphorylation. AB - Streptococcus mutans is the primary etiological agent of dental caries in man and other animals. This organism and other related oral streptococci use carbohydrates almost exclusively as carbon and energy sources, fermenting them primarily to lactic acid which initiates erosion of tooth surfaces. Investigations over the past decade have shown that the major uptake mechanism for most carbohydrates in S. mutans is the phosphoenolpyruvate (PEP)-dependent phosphotransferase system (PTS), although non-PTS systems have also been identified for glucose and sucrose. Regulation of sugar uptake occurs by induction/repression and inducer exclusion mechanisms in S. mutans, but apparently not by inducer expulsion as is found in some other streptococci. In addition, ATP-dependent protein kinases have also been identified in S. mutans and other oral streptococci, and a regulatory function for at least one of these has been postulated. Among a number of proteins that are phosphorylated by these enzymes, the predominant soluble protein substrate is the general phospho-carrier protein of the PTS, HPr, as had previously been observed in a variety of Gram positive bacteria. Recent results have provided evidence for a role for ATP dependent phosphorylation of HPr in the coordination of sugar uptake and its catabolism in S. mutans. In this review, these results are summarized, and directions for future research in this area are discussed. PMID- 2557098 TI - [Excitation-contraction coupling in the smooth muscle of the femoral artery under the effects of noradrenaline]. AB - Noradrenaline (5 x 10(-8) - 10(-5) M) induced a dose-dependent contraction of muscle strips from rabbit femoral artery. At concentrations higher than 10(-7) M noradrenaline evoked also a depolarization of smooth muscle cells due to an increase in sodium and/or chloride permeability of the membrane. Repolarization of the membrane to original level by inwardly applied current resulted in restoration of membrane resistance and partial relaxation of noradrenaline-evoked contraction. The same part of contraction was also blocked by verapamil. In calcium-free EGTA-containing solution noradrenaline induced only a small transient contraction. These findings indicate that noradrenaline-activated sodium (or chloride) permeability is voltage dependent. Noradrenaline evoked contraction is activated by calcium ions entered the cell through receptor operated and partly through voltage-operated calcium channels. PMID- 2557100 TI - [Increase in the number of alpha-2 adrenoceptors in blood platelets of patients with hypo-alpha-cholesterolemia]. PMID- 2557099 TI - [Adaptation to stress increases the resistance of isolated heart to damaging effects of Ca2+ due to optimization of Ca pump function in the sarcoplasmic reticulum]. AB - The adaptation of rats to repeated short-term immobilization stress prevents the calcium-induced arrhythmias and contracture on isolated heart. Obtained results indicate, that main role in this effect plays an increase of both Ca-transport efficiency in sarcoplasmic reticulum and its resistance to autolytic processes. PMID- 2557101 TI - [Effects of polymethylene derivatives of 4-aminopyridine on functional properties of hippocampal neurons]. AB - The effects of amiridin (9-amino-2,3,5,6,7,8-hexahydro-IH-cyclopenta(b) quinoline) and tacrine (1,2,3,4-tetrahydro-9-aminoacridine) on Schaffer collaterals--CAI field potentials were compared in rat hippocampal slice preparations. Similar dose-dependent increase in pop-spike amplitude was observed during slice perfusion with low concentrations of amiridin (5-50 microM) or tacrine (0.5-10 microM). This facilitation was not always fully reversible. The effect was accompanied by slight decrease in pop-EPSP amplitude suggesting membrane depolarization as a possible mechanism of pop-spike facilitation. Further increase in drug concentrations led to the depression and full blockade of pop-spike, that was associated with significant decrease in the pop-EPSP and fiber potential amplitudes. In contrast structurally related 4-aminopyridine evoked dose-dependent increase in both pop-EPSP and pop-spike amplitudes with all the concentrations tested (0.05-1000 microM), this facilitation was transformed into epileptiform response with 4-aminopyridine concentration about 500 microM. Possible mechanisms of drug actions on hippocampal neuron reactivity are discussed. It is suggested that amiridin might turn to be as effective as tacrine in symptomatic treatment of Alzheimer disease. PMID- 2557102 TI - [Effects of ethanol on the concentration of neuropeptides, ACTH and corticosterone during immobilization stress]. AB - It was shown, that stress increased the level of ACTH, beta-endorphin and corticosterone in the blood plasma of the rat. Injection of ethanol (1 g/kg) decreased the level of ACTH, but increased the levels of beta-endorphin in the rat subjected to immobilization stress. The immobilization lowered the levels of met-enkephalin in the striatum and medulla oblongata, but increased the content of neuropeptide in the adrenal glands. The concentration of leu-enkephalin and DSIP remained unchanged following the stress. Ethanol reversed the action of immobilization on the level of met-enkephalin in the striatum, but increased the content of DSIP in the thalamus. These results indicate that ethanol modified the activity of pituitary-adrenal-axis during stress and probably the stress protective action of ethanol partly performed with the involvement of DSIP. PMID- 2557103 TI - [Effects of peptide hydra morphogen on the contents of cyclic nucleotides in injured tissues]. AB - It is shown in experiments on mice that injection of Hydra Peptide Morphogen in doses of 30 micrograms/kg normalizes the content of cyclic nucleotides in the liver, disturbed as a result of injury. Hydra Peptide Morphogen did not change the content of cyclic nucleotides in the injured muscle. PMID- 2557104 TI - Protein C and protein S. PMID- 2557105 TI - Persistence of cytomegalovirus in peripheral blood from blood donors. PMID- 2557107 TI - Peripheral neuropathy and mesalazine. PMID- 2557106 TI - Risks of donor insemination. PMID- 2557108 TI - Unitary models of schizophrenia. PMID- 2557109 TI - Dementia and Parkinson's disease. AB - Recent research into the dementia of Parkinson's disease has exposed a complex area in which it has not always been possible to match clinical and pathological observations. Certain neuropsychological deficits result from a disruption of basal ganglia and frontal lobe interactions. These are unrelated to a global dementia, the prevalence of which exceeds twice that in the normal population. The associated pathological lesions comprise cortical pathology, either Alzheimer's disease or Lewy bodies, in combination with moderate degeneration of the subcortical, cholinergic, basal nucleus of Meynert. PMID- 2557110 TI - Cerebral vascular transit time in Alzheimer's disease and Korsakoff's psychosis and its relation to cognitive function. AB - The cerebral vascular transit time of 17 patients with pre-senile dementia of the Alzheimer type (ATD), nine abstinent patients with alcoholic Korsakoff's psychosis (KOR), and ten age-matched controls was determined by the bolus intravenous injection of pertechnetate. A gamma camera was used to estimate the median transit time (MTT) of the radioactive bolus in a planar (non-tomographic) projection normal to the vertex. The spread of the bolus arriving at the aortic arch was measured independently by a single external detector over the chest, and correction made for the transit time of this input function in calculating the net MTT for the head. Both ATD and KOR groups showed lengthened net MTTs, compatible with reduced cerebral blood flow, and which were correlated with reduced cognitive function. It is concluded that the method employed gives a simple, inexpensive estimate of function-related blood flow to the brain in pre senile dementia. PMID- 2557111 TI - The near-death experience. PMID- 2557113 TI - Lipids in breast carcinogenesis. AB - Excess dietary fat has been identified as a risk factor in the development of human breast carcinoma. However, the quality of fat may be more important than the overall quantity. We have studied the growth of human MCF7 breast carcinoma xenografts in athymic mice treated with dietary supplements of N-6 and N-3 series essential fatty acids given as natural preparations of evening primrose oil and fish oil. Olive oil and normal laboratory diet lacking the essential fatty acids served as controls. Animals treated with essential fatty acids developed tumours which were significantly smaller than both control groups (Mann-Whitney U test, P less than 0.001). Median tumour weights according to diet were: evening primrose oil, 133 mg; fish oil, 70 mg; olive oil, 212 mg; and control, 270 mg. Nutritional intervention to increase the proportion of essential fatty acids in the diet may have a role in the management of breast carcinoma. PMID- 2557112 TI - New radionucleotide method of visualizing subclavian vein occlusion following temporary venous access. PMID- 2557114 TI - Tumour necrosis factor. AB - Tumour necrosis factor, TNF, possesses the attributes of a typical cytokine, being a pleiotropic cell regulatory protein whose activity is largely determined by the cell type to which it binds and the presence of other protein regulators. Thus TNF can be a growth factor, cytotoxin, cytostatic agent or inducer of differentiation. This cytokine is also an important inflammatory mediator regulating the activity of neutrophils, eosinophils, T and B lymphocytes and modulating the properties of the vascular endothelium. All of these actions may play a role in the ability of TNF to induce necrosis of experimental animal tumours, but the potential of this cytokine in therapy of human cancer is uncertain. The increasing evidence for a role for TNF in the pathophysiology of acute and chronic disease makes the use of specific TNF antagonists an equally interesting therapeutic possibility. PMID- 2557115 TI - Transforming growth factors beta. AB - Since its discovery as a factor able to induce phenotypic transformation with transforming growth factor alpha, transforming growth factor beta (TGF beta) has been found to have unexpectedly widespread and frequently, seemingly paradoxical effects on a variety of cell types. Thus, a significant property of TGF beta activity is the ability to inhibit cellular growth and transformation. More recently, further important potential roles have also been found for TGF beta including the control of embryonal development, cellular differentiation, hormone secretion and immune function. Multiple forms of both TGF beta and TGF beta receptors are now known to exist, which may generate a complex and diverse range of interacting signals. The multifunctional actions of the TGF beta family are frequently defined in concert with other extracellular factors and much still has to be understood concerning these synergistic processes. PMID- 2557116 TI - Platelet-derived growth factors: a family of isoforms that bind to two distinct receptors. AB - Platelet-derived growth factor (PDGF) is a mitogen for connective tissue cells and occurs as disulphide-bonded homodimers or heterodimers of related polypeptide chains. Recent data indicate that the isoforms have different functional activities due to the fact that they bind with different affinities to two distinct receptor types. The frequent expression of PDGF and PDGF receptors in normal as well as transformed cells, suggests roles for PDGF in autocrine and paracrine stimulation of cell growth in vivo. PMID- 2557117 TI - Neuropeptides as growth regulators. AB - The classical role of neuropeptides as fast-acting neurohumoral signallers has recently been challenged by the discovery that many neuropeptides are also growth factors stimulating slow-acting mitogenesis. Their mechanisms of action have been studied in cell culture, and their cell-surface receptors have been characterized pharmacologically using agonists and antagonists. We describe the mitogenic effects of bombesin, vasopressin, bradykinin and vasoactive intestinal peptide in murine fibroblasts. We suggest that the receptors for bombesin, vasopressin and bradykinin have more than one binding site, permitting modulation of transmitted signals. As these neuropeptide receptors share the ability to mobilize intracellular Ca2+, their common domain may be essential to G-protein coupling. PMID- 2557118 TI - Signal transduction pathways in mitogenesis. AB - Quiescent cells arrested in the Go phase of the cell cycle can be stimulated to divide by polypeptide growth factors, pharmacological agents and neuropeptides which exhibit potent synergistic effects. Bombesin-like peptides are providing valuable model mitogens to elucidate the signalling pathways leading to mitogenesis. These peptides stimulate rapid increases in ionic fluxes, inositol polyphosphate formation, activation of protein kinases and expression of proto oncogenes. A comparison of these early molecular events with those evoked by other growth factors indicate the existence of multiple signal-transduction pathways. We propose that stimulation of cell proliferation by single or multiple factors results from the activation of separate signal-transduction pathways that cooperate to elicit the complete set of molecular events leading to mitogenesis. PMID- 2557119 TI - Growth factor receptors. AB - The initial interaction of growth factors with their target cells is mediated by specific high affinity cell surface receptors. The structural and functional diversity found in growth factors is more limited at the receptor level where receptors for structurally and functionally distinct factors seem to share structural motifs and employ a restricted number of signal transduction and second messenger systems. In many cases the receptors for particular factors remain to be characterized, but rDNA techniques offer the hope that all receptors can eventually be defined in structural detail. As yet understanding of the signal transduction mechanisms remains limited but when unravelled this may offer routes for artificial modulation of growth factor action. PMID- 2557120 TI - Structure-function relationships of growth factors and their receptors. AB - The primary amino acid sequences of several receptor tyrosine kinases have recently made it possible to deduce similarities in the molecular organization of these large multidomain proteins. This has allowed a classification of these receptors into three groups (see Waterfield this Issue and for review in Ref.1). Class I includes the EGF receptor and the neu proto-oncogene, Class II includes the insulin and insulin-like growth factor 1 (IGF-1) receptors, and Class III the platelet derived growth factor (PDGF) and the colony stimulating factor 1 (CSF-1) receptors. The conformation of the ligands for the Classes I and II receptors have been defined using X-ray diffraction, 2-D nuclear magnetic resonance (NMR) and knowledge based modelling procedures. It seems that the ligands are more diverse in sequence than the receptor tyrosine kinases so they cannot be classified as rigorously. However, certain features are common to all growth factors (so far defined) which form compact, globular structures and this allows a discussion of possible interactions between the ligand and receptor; but in the absence of a molecular structure for any of the receptors, we can only review biochemical evidence and deductions from predictive and modelling studies. Various models for the signal transduction process are discussed in the light of current work on receptor interactions. PMID- 2557121 TI - Polyneuropathy in feline Niemann-Pick disease. AB - Two related cats, aged 5 months and 7 months, and 1 unrelated cat, aged 4 months, presented with signs of a progressive neuromuscular disease. Detailed electrophysiological studies suggested a primary demyelinating polyneuropathy, which was confirmed by muscle and nerve biopsies and on necropsy examination. Light and electron microscopic findings indicated a lysosomal storage disease, which was diagnosed as sphingomyelinase deficiency (Niemann-Pick disease) by enzyme analysis and lipid fractionation, although significant biochemical differences existed between the 2 related cats and the third cat. Several lines of evidence suggest that these 2 related cats were affected with a variant of type A Niemann-Pick disease, whereas cat 3 represented classic Niemann-Pick disease type A. PMID- 2557122 TI - Disappearance of the mu-opiate receptor patches in the rat neostriatum following lesioning of the ipsilateral nigrostriatal dopamine pathway with 1-methyl-4 phenylpyridinium ion (MPP+): restoration by embryonic nigral dopamine grafts. AB - Using in vitro autoradiography, this study examined the binding of the selective mu-opiate receptor radiolabelled ligand, [3H]Tyr-D-Ala-Gly-Me-Phe-Gly-ol ([3H]DAGO) to the striatal sections of rats with long-term unilateral lesions of the nigrostriatal dopamine (DA) pathway induced by 1-methyl-4-phenylpyridinium ion (MPP+) and in animals bearing embryonic DA grafts implanted into the DA depleted striatum. In the ipsilateral striatum of MPP+-lesioned animals, there was a complete disappearance of the mu-opiate receptor patches as well as the subcallosal streak. The normal pattern of mu-binding sites in the patches reappeared following reinnervation of the DA-depleted striatum by the DA-grafts. These findings suggest that mu-opiate receptor patches in the striatum are localised on nigrostriatal DA afferent terminals. However, it is possible that trans-synaptic or postsynaptic changes also contribute to the profound alterations in striatal mu-opiate binding patterns revealed in this study. PMID- 2557124 TI - Immunocytochemical localization of the GABAA/benzodiazepine receptor in the guinea pig cochlear nucleus: evidence for receptor localization heterogeneity. AB - Immunocytochemistry with a monoclonal antibody against the GABAA/benzodiazepine receptor showed labeled axo-dendritic synapses in the anteroventral cochlear nucleus. In the dorsal cochlear nucleus, label was seen apposing both axo-somatic and axo-dendritic terminals. The results suggest a heterogeneous distribution of GABA receptors, together with a possible segregation of receptor subtypes between somata and dendrites in certain neurons. The presence of cytoplasmic labeling in some neurons might reflect a higher receptor turnover rate in these neurons. PMID- 2557123 TI - Age-related differences in the sensitivity of rats to a selective sigma ligand. AB - Differences in sigma binding parameters and behavioral responses to injections of a selective sigma ligand were found in adult rats of different ages. Middle-aged rats (5-6 months old) had fewer sigma binding sites and sites with lower affinity for [3H]di-o-tolylguanidine than young adult animals (2-3 months old). The older animals also exhibited a decreased behavioral response to the selective sigma ligand, di-o-tolylguanidine (DTG). Unilateral microinjection of DTG into the substantia nigra of rats produced fewer net contralateral turns in middle-aged animals, compared to younger rats. Likewise, the postural changes produced by unilateral microinjection of DTG into the red nucleus were less pronounced in the older animals. These data suggest that changes in the number and affinity of sigma binding sites may affect movement and posture as an organism ages. PMID- 2557125 TI - Localization of cyclic adenosine monophosphate in the teleost retina: effects of dopamine and prolonged darkness. AB - Localization of cyclic adenosine monophosphate (cAMP) in the white perch retina was carried out with immunohistochemical and autoradiographic methods. Following exposure to dopamine or prolonged darkness, cAMP staining was observed by immunohistochemistry in the distal part of the inner nuclear layer, i.e. in the horizontal cells. After exposure to dopamine, increased levels of cAMP were also observed by autoradiography in many horizontal cells. Finally, increased levels of cAMP staining were observed immunohistochemically following incubation with dopamine in all types of cone-related horizontal cells that had been isolated and maintained in culture. PMID- 2557126 TI - The coupling of muscarinic receptors to hydrolysis of inositol lipids in human neuroblastoma SH-SY5Y cells. AB - Carbachol (CCh)-stimulated hydrolysis of inositol lipids in human neuroblastoma SH-SY5Y cells was systematically characterized in parallel with the carbachol effects on cAMP formation. Carbachol concentration-dependently induced the hydrolysis of inositol lipids and formation of [3H]IP3, [3H]IP2 and [3H]IP1 in these cells labeled with [3H]inositol. The maximal amount of [3H]IP1 accumulated in the presence of 10 mM LiCl was about 50-fold above the basal level. The EC50 value of CCh was 14 microM. The muscarinic antagonists atropine, pirenzepine and 11-[[2-(diethylamino)methyl]-1-piperidinyl]acetyl]-5,11-dihydro- 6H-pyrido [2,3 b] (1,4)-benzodiazepine-6-one (AF-DX 116) competitively inhibited CCh-induced [3H]IP1 accumulation. The functional inhibition constants (converted from the pA2 values) were 0.24, 8.1 and 470 nM, respectively. These values are in good agreement with the inhibition constants of these drugs from antagonist/[3H]pirenzepine studies using intact cells. Forskolin, adenosine and PGE1 stimulated cAMP formation in this cell line. Morphine decreased PGE1-induced cAMP formation as well as the basal cAMP formation. However, CCh did not stimulate or inhibit the basal cAMP formation. Also, CCh did not have any effects on the adenosine and PGE1-induced cAMP formation in these cells. These data suggest that muscarinic M1 receptors are coupled to the hydrolysis of inositol lipids and not to the adenylate cyclase system in human neuroblastoma SH-SY5Y cells. PMID- 2557127 TI - Imipramine alters beta-adrenergic, but not serotonergic, mediated responses in rat hippocampal pyramidal cells. AB - Imipramine, a tricyclic antidepressant, acts acutely to block the reuptake of serotonin (5-HT) and norepinephrine (NE). However, imipramine's action as an antidepressant takes several weeks to develop. This study investigated acute and chronic effects of imipramine on intracellularly-recorded responses mediated by 5 HT and beta-adrenergic receptors on pyramidal cells from area CA1 of rat hippocampal slices maintained in vitro. Addition of 10 microM imipramine in the perfusion medium sinistrally shifted the 5-HT1A concentration-response curve for membrane hyperpolarization and the 5-HT concentration-response curve for the reduction in the amplitude of the slow afterhyperpolarization (AHP) elicited by a train of action potentials. After two weeks of treatment with imipramine (10 mg/kg daily i.p. injections or s.c. osmotic mini-pumps) the responses to 5-HT were not altered. In contrast the concentration-response curve for the beta adrenergic mediated reduction in AHP amplitude was significantly altered; there was a reduction in Emax and a log unit dextral shift in EC50. There was no change in the concentration-response curve for the beta-adrenergic mediated depolarization. These data are in agreement with previous biochemical results reporting a decrease in beta-adrenergic receptor mediated stimulation in adenylyl cyclase and down-regulation of beta-receptor in cortex and hippocampus. These findings suggest that a consequence of long-term imipramine treatment is a decrease in the augmentation of cell excitation normally produced by beta adrenergic receptor stimulation. PMID- 2557128 TI - Chick myotubes in culture express high-affinity receptors for calcitonin gene related peptide. AB - Calcitonin gene-related peptide (CGRP) is a 37-amino acid neuropeptide that is expressed by many neurons of the vertebrate nervous system, including motoneurons of many species. It has been detected immunohistochemically in both cell bodies and motor terminals of motoneurons, suggesting that it may play a role at the neuromuscular junction. In support of this idea, CGRP has been shown to produce a variety of effects on cultured myotubes and muscle explants, including elevation of cAMP levels, increase in cell-surface acetylcholine receptor (AChR) numbers, increase in AChR alpha-subunit mRNA transcript levels, alterations in contractile responses, alterations in the physiological properties of AChRs, and inhibition of insulin-induced changes in glycogen metabolism. CGRP binding sites have been detected in many tissues, but have not yet been demonstrated directly on muscle cells. Here we report that chick myotubes in culture express high-affinity binding sites for CGRP (Kd approximately 2-4 x 10(-10) M). In view of the known biological effects of CGRP on myotubes, we believe that these binding sites represent CGRP receptors. They are uniformly distributed over the surface of myotubes, and we have found no evidence of clustering in culture, in contrast to AChRs. We have found no evidence for more than one class of receptors. PMID- 2557129 TI - Influences of external Ca2+ on the GABA-induced chloride current and the efficacy of diazepam in internally perfused frog sensory neurons. AB - The effects of extracellular Ca2+ on the gamma-aminobutyric acid (GABA)-induced Cl- current and the efficacy of diazepam in the facilitation of GABA response were studied in frog isolated sensory neurons, using a 'concentration clamp' technique which combines a suction pipette (internal perfusion and voltage clamp) and a rapid drug application system. When nominal Ca2+-free external solution was changed to the solution containing 2 mM Ca2+, the response elicited by 1 x 10(-5) M GABA was reduced by about 40% of the control obtained in nominal Ca2+-free solution. The dose-response curve for GABA was shifted to the right without affecting the maximum response. It can be suggested that the application of external Ca2+ modulates the affinity of the GABA receptor to its agonist, GABA. Diazepam at the concentration of 3 x 10(-6) M shifted the dose-response curve for GABA to the left without changing the maximum response with or without external Ca2+. However, the augmentatory action of diazepam on the GABA response was reduced in the presence of external Ca2+. Possible mechanisms for inhibitory action of external Ca2+ on the GABA-gated response and the reduced efficacy of diazepam are discussed. PMID- 2557130 TI - Glycine reverses 7-chlorokynurenic acid-induced inhibition of [3H]MK-801 binding. AB - 7-Chlorokynurenic acid (7-Cl KYNA) has been reported to attenuate N-methyl-D aspartate (NMDA) receptor functioning by a potent and selective inhibitory action mediated at the strychnine-insensitive glycine recognition site of the NMDA complex. Here we report that 7-Cl KYNA dose-dependently inhibits [3H]MK-801 binding to the PCP receptor, and that this effect is reversed by addition of glycine. Since [3H]MK-801 binding is a measure of channel activation, our results are consistent with the hypotheses that 7-Cl KYNA exerts its NMDA receptor antagonism by acting at the glycine site, and that activation of the glycine site is required for NMDA channel activity to occur. PMID- 2557131 TI - Localization of cGMP in the cerebellum of the adult rat: an immunohistochemical study. AB - The localization of cGMP in the cerebellum of the adult rat after fixation with formaldehyde was studied with an antibody raised against a cGMP-formaldehyde thyroglobulin conjugate. Three different protocols were used: (1) in vitro incubation of 300 microns cerebellar slices followed by fixation and cryostat sectioning; (2) in vitro incubation of 100 microns cerebellar slices followed by fixation with no further sectioning; (3) perfusion fixation of the anesthetized rat followed by vibratome sectioning. All 3 protocols gave essentially the same results: cGMP-immunoreactivity was found predominantly in Bergmann fibers in the molecular layer, in Bergmann cell bodies in the Purkinje cell layer (but not in Purkinje cells), and in astroglial cells in the granular layer. PMID- 2557132 TI - Calcium-dependent chloride currents elicited by injection of ethanol into Xenopus oocytes. AB - Electrophysiological techniques were used to study the response of native Xenopus laevis oocytes to intracellular injection and bath application of ethanol. Injection of ethanol produced dose-dependent transient inward currents accompanied by large current fluctuations, with estimated intracellular concentrations ranging from 10 to 300 mM. The response duration varied between 2 and 15 min, with an onset delay of 2-15 s. The inward current sometimes consisted of a fast and a slow component. Bath application of equivalent concentrations elicited similar but considerably smaller responses. The current showed a reversal potential of -20 +/- 10 mV, corresponding to an increase in chloride permeability. The response was eliminated in the presence of low chloride saline and was blocked by the chloride channel inhibitors SITS and DIDS. Ethanol responses were inhibited by the intracellular injection of the calcium chelator EGTA and were unaffected when the extracellular calcium was lowered. It is concluded that ethanol injection into Xenopus oocytes elicits a release of calcium from intracellular stores, which then activates an increased membrane permeability to chloride. PMID- 2557133 TI - Distribution of glucocorticoid receptor-like immunoreactivity in the brain, and its relation to CRF and ACTH immunoreactivity in the hypothalamus of the japanese quail, Coturnix coturnix japonica. AB - Monoclonal antibody (MAb49) against the rat liver glucocorticoid receptor was used to evaluate glucocorticoid receptor (GR) immunoreactive structures in the brain of the japanese quail, Coturnix coturnix japonica. Using the avidin-biotin technique, the immunoreaction was present in the nerve cell nuclei in intact male birds. High density of glucocorticoid receptor-like immunoreactivity was found in the tubero-infundibular area, nucleus paraventricularis, posteromedialis and lateralis hypothalami, lateral septum and in some brainstem nuclei. Cerebellar cortex was also immunopositive. In contrast to mammals, no immunoreactive cell nuclei were found in the hippocampal region. The glucocorticoid receptors were colocalized with adrenocorticotropin (ACTH) immunoreactivity in the tubero infundibular region, while corticotropin releasing factor (CRF) positive cells in the paraventricular nucleus did not contain glucocorticoid receptor-like immunoreactivity in their cell nuclei. PMID- 2557134 TI - Cerebral metabolic effects of sigma ligands in the rat. AB - The 2-deoxy-D-[1-14C]glucose method was used to study the effects of sigma-type drugs, BMY 14802, (+)-pentazocine and BW 234U (rimcazole), on cerebral metabolism in 44 male Fischer 344 rats. Drug effects were observed in epithalamic, methathalamic, hypothalamic and mesencephalic regions, as well as in cranial nerve nuclei, the cerebellum, and the medulla oblongata. BMY 14802 and (+) pentazocine increased local cerebral glucose utilization (LCGU) in areas that generally did not overlap; BW 234U administered 30 min before the radiotracer decreased LCGU. Most of the areas that showed changes in LCGU are known to contain sigma receptor sites. Differences in pharmacological properties, including effects on neuronal electrical excitability, may have resulted in the different distributions and effects of the drugs on LCGU. The present findings did not discriminate the compounds as agonists or antagonists for sigma receptor sites. However, the 2-deoxy-D-[1-14C]glucose method appeared useful in delineating the distribution of CNS responses to sigma drugs in the rat. PMID- 2557135 TI - [Use of electron paramagnetic resonance spectroscopy spin labels in studying the properties of biological membranes]. AB - Electron paramagnetic resonance (EPR) spectroscopy of spin labels has become a widely spread method in biochemistry, molecular biology, and medicine. After a brief view on the structure and functions of biological membranes, basic principles of EPR and particularly of the spin labeling technique are presented. Applications of this technique are demonstrated on the study of various properties of biological membranes, such as phase transition of lipids, properties of different membrane parts, permeability of membranes, inner volumes of liposomes and cells, membrane potentials, lateral diffusion, lipid-protein interaction, and determination of oxygen concentration. As an example the results of a study concerning the effect of chlorpromazine and its derivatives on the dynamics and lipid-protein interaction in synaptosomes are presented. PMID- 2557137 TI - [The quantity and distribution of specific glycoproteins in gastric carcinoma]. AB - 42 tissue samples of gastric carcinoma were stained by HE, PAS and AB, and treated by Immuno-histochemistry ABC with 7 kinds of Lectin to study the meaning of the changes of glycoprotein in their different types and different sites of the gastric mucosa. The binding indexes of Dolichos biflorus agglutinin (DBA) (6.81) and Soybean agglutinin (SBA) (7.02) were strong (P less than 0.01) in mucus carcinoma. In undifferentiated glandular carcinoma, Wheat germ agglutinin (WGA) was high (6.98, P less than 0.05). WGA staining in the tumor of invaded nerves or vessels was strong. The reactions of Ricinus communis agglutinin (RCA) and WGA in most tumors were stronger than those of the other Lectins. Some positive areas of Lectin binding were negatively presented after treating with PAS and AB methods. The results indicated that Lectin binding was more sensitive than classical histochemical methods. The changes of DBA and SBA showed increased N-acetyl-D-galactosamine in mucus carcinoma. The changes of WGA suggested that the increased sialic acid reaction was related to the differentiation, invasion and metastasis of tumor. PMID- 2557136 TI - [An uncommon form of thymidine kinase in breast cancers]. AB - An unusual form of thymidine kinase has been found in cytosols from breast cancers. It differs from the adult and fetal isoenzymes of thymidine kinase in so far as its activity could not be isolated from that of thymidylate kinase. Following sucrose density gradients, electrofocusing, ion exchange chromatography or molecular sieving, both activities appeared as a single peak. These facts suggest that they could be attributed to a single multifunctional protein. PMID- 2557138 TI - [A study of ricinus communis agglutinin I receptors in liver cancer tissues]. AB - Ricinus communis agglutinin I(RCAI) receptors in 25 cases of hepatocellular carcinoma and 6 cases of intrahepatic cholangiocellular carcinoma were immunohistochemically localized by avidin-biotin-peroxidase complex (ABC) method. In the meantime, RCAI receptors in normal and cirrhotic liver tissues were also observed as controls. The results showed that there were many irregularly distributed RCAI receptors in HCC in forms of dispersed dots, even or localized lumpy stainings. The receptors in most intrahepatic cholangiocellular carcinomas were distributed in a polar form. However, the distribution of RCAI receptors in hepatocytes of normal, cirrhotic and precancerous liver tissues was band-like. It is suggested that the distribution of RCAI receptors in the cells might be helpful to the diagnosis of hepatoma and to the differentiation of benign from malignant hyperplasia. PMID- 2557139 TI - [Heterologous regulation of adrenergic receptors]. PMID- 2557140 TI - Immediate hypersensitivity reactions in the guinea-pig conjunctiva: studies with a second-generation leukotriene D4 receptor antagonist, MK-571. AB - The role of leukotriene D4 (LTD4) as a mediator of immediate hypersensitivity reactions in the guinea-pig conjunctiva was examined using a potent, second generation LTD4 receptor antagonist, MK-571 (also known as L-660,711). The microvascular permeability changes in the guinea-pig conjunctiva following challenge with either LTD4 or antigen were measured through accumulation of intravenously administered 99mtechnetium-labeled albumin. Topical application of MK-571 (up to 2 h pretreatment) significantly inhibited the conjunctival responses to LTD4 (ED50 of 18-60 ng/eye) but not to histamine. The responses to a single topical antigen challenge in ovalbumin-sensitized guinea pigs were significantly inhibited (44%) by topical treatment with MK-571, in contrast to the lack of effect previously observed with prototypic antagonists. The inhibitory effects of MK-571 did not involve an action on conversion of [3H]LTC4 to LTD4 and LTE4. Following a second antigen challenge (24 h after the first), MK 571 inhibited the resultant permeability changes by 78%. Specific histamine H1 and H2 antagonists similarly inhibited the responses to the first and second challenges (63 and 74%, respectively). The present study suggests that LTD4 is involved in conjunctival hypersensitivity reactions and that potent LTD4 receptor antagonists may be of therapeutic value in the treatment of allergic conjunctivitis. PMID- 2557141 TI - Effects of GTP analogs and dithiothreitol on the binding properties of the vascular vasoactive intestinal peptide receptor. AB - Previous studies have demonstrated a specific vascular receptor for the neurotransmitter peptide, vasoactive intestinal peptide (VIP), and have suggested that the receptor is positively coupled to vascular adenylate cyclase. The present study addressed the questions whether the vascular VIP receptor is subject to regulation by guanine nucleotides and whether a disulfide reducing agent, dithiothreitol, would perturb the binding function of the vascular VIP receptor. Guanosine triphosphate (GTP) and its non-hydrolyzable analogs, guanylyl imidodiphosphate (Gpp(NH)p) and guanosine-5'-O-(3-thiotriphosphate) (GTP-gamma S), increased the rate of dissociation of radiolabeled VIP from arterial receptors in a concentration-dependent manner. GTP-gamma-S increased the equilibrium dissociation constant (KD) of the high affinity vascular VIP binding site, a result consistent with decreased high affinity binding of VIP induced by GTP-gamma-S. These results are consistent with a regulatory role for guanine nucleotides in the function of the vascular VIP receptor. The disulfide reducing agent, dithiothreitol, caused a decrease in specific binding of radiolabeled VIP. Upon Scatchard analysis the effect of dithiothreitol was characterized by an increase in the KD and a decrease in the maximum number of binding sites (Bmax) of the high affinity binding site. These results suggest that disulfide bonds are important for ligand binding to vascular VIP receptors. The sulfhydryl alkylating agents, N-ethylmaleimide and iodoacetamide, had minimal effects on radioligand binding. PMID- 2557142 TI - Postnatal development of adrenergic responsiveness in the rabbit heart. AB - It is uncertain how changes in the beta-adrenoceptor population influence the contractility of developing heart. To resolve this we have examined postnatal developmental changes in the adrenergic responsiveness of the rabbit heart. The inotropic effect of isoproterenol on isolated left ventricular papillary muscles from rabbits aged 3, 21, and 90 days was compared with the relative number of beta-adrenoceptors at each age measured using [3H]dihydroalprenolol ([3H]DHA) as the specific ligand. The maximum tension developed in response to isoproterenol increases from 37 +/- 7 to 175 +/- 33% above control twitch tension between 3 and 21 days of age; this is followed by a decrease to 68 +/- 12% in the young adult. During this period of development, there is a decline in EC50 towards increased sensitivity. These differences are partially accounted for by an increase in the numbers of specific [3H]DHA binding sites from 17.3 +/- 2.3 to 56.6 +/- 9.9 fmol/mg wet tissue weight from 3 to 21 days, and a subsequent decrease to 32 +/- 4.5 fmol/mg tissue in the young adult. The proportionally larger increase in contractility compared with the number of beta-adrenoceptor binding sites during the first 3 weeks of life is discussed in terms of the developmental changes in the efficacy of coupling between receptor occupancy and contraction. PMID- 2557143 TI - Caffeine treatment inhibits drug-induced calcium release from sarcoplasmic reticulum and caffeine contracture but not tetanus in frog skeletal muscle. AB - Effects of pretreatment with caffeine on Ca2+ release induced by caffeine, thymol, quercetin, or p-chloromercuriphenylsulfonic acid (pCMPS) from the heavy fraction of sarcoplasmic reticulum (SR) were studied and compared with those effects on caffeine contracture and tetanus tension in single fibers of frog skeletal muscle. Caffeine (1-5 mM) did induce transient Ca2+ release from SR vesicles, but subsequent further addition of caffeine (10 mM, final concentration) induced little Ca2+ release. Ca2+ release induced by thymol, quercetin, or pCMPS was also inhibited by pretreatment with caffeine. In single muscle fibers, pretreatment with caffeine (1-5 mM) partially reduced the contracture induced by 10 mM caffeine. However, tetanus tension was almost maximally induced by electrical stimulus in caffeine-treated fibers. These results indicate that SR, which becomes less sensitive to caffeine, thymol, quercetin, or pCMPS by pretreatment with caffeine, can still respond to a physiological signal transmitted from transverse tubules. PMID- 2557144 TI - The role of aberrant hypothalamic opiatergic function in generating polycystic ovaries in the rat. AB - Treatment of adult female rats with estradiol valerate produces an intractable hypothalamic impairment that ultimately results in anovulatory acyclicity and polycystic ovaries. Evidence from our laboratory suggests that the hypothalamic impairment compromises regulation of the endogenous opioid system engendering a persistent opiatergic suppression of gonadotropin-releasing hormone secretion, which is subsequently reflected in a chronically low pituitary content of gonadotropin-releasing hormone receptors. If such is the case, inhibition of opiatergic transmission should improve the gonadotropin-releasing hormone pattern resulting in an improvement in the pituitary content of gonadotropin-releasing hormone receptors, and in an amelioration of the polycystic condition. We, therefore, treated rats with the polycystic ovarian condition, with daily injections of naltrexone. Within 1 week, there was a significant increase in the pituitary content of gonadotropin-releasing hormone receptors and a marked improvement in ovarian morphology, indicating that the hypothalamic opiatergic system is chronically active, and contributes significantly to the polycystic ovarian condition. PMID- 2557145 TI - Reduced neuroexcitatory effect of domoic acid following mossy fiber denervation of the rat dorsal hippocampus: further evidence that toxicity of domoic acid involves kainate receptor activation. AB - Domoic acid, an excitatory amino acid structurally related to kainic acid, has been shown to be responsible for the severe intoxication presented, in 1987, by more than one hundred and fifty people having eaten mussels grown in Prince Edward Island (Canada). Unitary extracellular recordings were obtained from pyramidal neurons of the CA3 region of the rat dorsal hippocampus. The excitatory effects of microiontophoretic applications of domoic acid and of the agonists of the two other subtypes of glutamatergic receptors, quisqualate and N-methyl-D aspartate, were compared on intact and colchicine-lesioned sides. Similar to what has been previously found for kainate, the colchicine lesion of the mossy fiber projections induced a 95% decrease of the neuronal responsiveness to domoic acid, whereas the effect of quisqualate was unchanged and that of N-methyl-D-aspartate was only slightly decreased. These results provide further electrophysiological evidence that domoic acid is a potent agonist of kainate receptors and that it may produce its neuroexcitatory and neurotoxic effects, in the hippocampal CA3 region, through activation of kainate receptors located on the mossy fiber terminals. PMID- 2557146 TI - Positive inotropic effect of the thromboxane analog U-46619 on guinea pig left atrium: mediation by specific receptors and association with increased phosphoinositide turnover. AB - U-46619, a stable epoxymethano analog of thromboxane A2 elicited a direct positive inotropic effect on guinea pig left atrium paced at a constant rate (EC50 = 2.5 nM). This novel observation contrasts with previous reports of a decrease in myocardial contractility by thromboxane mimetic compounds in coronary perfused preparations, an action recognized as secondary to vasoconstriction. The positive inotropic effect of U-46619 was competitively antagonized by the specific thromboxane receptor blocker L-655,240 (pA2 = 8.02; identical to that reported in smooth muscle), but was unaffected by blockers of alpha 1-, beta 1-, and H1-receptors and by cyclooxygenase and lipoxygenase inhibitors. Increased tissue levels of inositol phosphates, but not cAMP, were associated with the positive inotropic action of U-46619, in analogy to the actions of alpha 1- and H1-receptor agonists. However, the inotropic effect of U-46619 and the concomitant increase in phosphoinositide breakdown were both selectively antagonized by L-655,240. Thus, U-46619 acts on specific thromboxane receptors in guinea pig left atrium and elicits a positive inotropic effect that probably results from an increase in phosphoinositide metabolism. PMID- 2557147 TI - Beta 2-adrenoceptor response in the rat uterus at the end of gestation and after induction of labor with RU 486. AB - To study the relationship between the progesterone environment and beta adrenoceptors in the myometrium, rats were treated with the antiprogestin RU 486 (10 mg per rat) at 08:30 h on day 21 of gestation. Under these conditions, more than 60% of animals delivered within 24 h after this treatment, while none of the control animals delivered within the same time period. beta-Adrenoceptors were identified using the radiolabeled antagonist (-)-[125I] iodocyanopindolol. The density (Bmax approximately 33-45 fmol/mg protein) and the affinity (KD approximately 0.105-0.106 nM) were not changed (during the late stages of gestation) in RU 486 treated rats compared with control rats. These results were correlated with the relaxation of longitudinal and circular strips of myometrium placed in high KC1 medium and exposed to beta-adrenoceptor agonists. The adrenoceptors implicated in the relaxation of myometrial strips were mainly of the beta 2-subtype. There was no difference in their affinity between control and RU 486 treated rats. Mean pA2 values were 8.46 for propranolol and 8.27 for ICI 118-551 against salbutamol. Altogether these results indicate in the rat that the blockade of the action of progesterone at its receptor site by RU 486 did not modify either the affinity or the sensitivity of beta 2-adrenoceptors in the myometrium, although it induced parturition. PMID- 2557148 TI - Characterization of alpha-adrenoceptor subtypes by [3H]prazosin and [3H]rauwolscine binding to canine venous smooth muscle membranes. AB - Postsynaptic alpha-adrenoceptor subtypes were studied using [3H]prazosin and [3H]rauwolscine binding to plasmalemma-enriched microsomal fractions isolated from dog saphenous veins and mesenteric veins. Both radioligands showed saturable binding consistent with the presence of a single homogeneous binding site in each case, based on Scatchard analysis. The Kd values of [3H]prazosin and [3H]rauwolscine, calculated from kinetic studies were similar to those from equilibrium binding data in both venous muscle membranes. The microsomal membranes of dog saphenous vein and mesenteric vein contained about a fourfold higher density of the high affinity [3H]rauwolscine binding sites than those for [3H]prazosin binding. In competition studies, IC50 values for displacement of rauwolscine or prazosin suggested that the sites of interaction for the antagonists prazosin and rauwolscine were independent. Phenylephrine, a functionally selective alpha-adrenoceptor agonist, competed with a similar IC50 value for the specific binding sites of [3H]prazosin and [3H]rauwolscine; but B HT 920, a functionally selective alpha 2-adrenoceptor agonist, competed for [3H]rauwolscine and [3H]prazosin binding with distinctly different IC50 values. Our data show the existence of two populations of alpha-adrenoceptor antagonist binding sites in the plasma membranes of dog saphenous vein and mesenteric vein, and raise the question whether agonist selectively depends on different affinities or on differential efficacies at one or two sites. PMID- 2557149 TI - Does in vitro activation of postsynaptic alpha 2-adrenoceptor utilize intracellular Ca2+ for contraction in dog saphenous vein? AB - Dog saphenous vein spiral strips were employed to determine whether an intracellular source of Ca2+ is used for contraction upon activation of the alpha 2-adrenoceptor by B-HT 920 in Ca2+-free Krebs solution containing 50 microM EGTA. The studies were carried out in parallel with the activation of the alpha 1 adrenoceptor by phenylephrine (Phe) under the condition that B-HT 920 (10(-5) M) and Phe (2 x 10(-6) M) gave rise to a similar level of responses in Ca2+ containing Krebs solution. A similar level of responses to these agonists at equieffective concentrations in Ca2+-free medium were also observed. Such responses to Phe and B-HT 920 were inhibited by 10(-7) M rauwolscine and 10(-7) M prazosin, respectively, and were not affected by 10(-7) M nifedipine or 5 mM Mn2+. The responses to B-HT 920 (10(-5) M) and submaximal concentration of Phe (2 x 10(-6) M) in Ca2+-free medium were additive. However, if the vascular strips were first contracted maximally with 10(-4) M Phe in Ca2+-free medium to deplete the intracellular Ca store, subsequent addition of B-HT 920 failed to induce additional response. Our results strongly suggest that activation of alpha 2 adrenoceptor in dog saphenous vein in Ca2+-free medium indeed utilizes intracellular Ca2+ for contraction. We also found that the failure of earlier studies to demonstrate the contractile effects of B-HT 920 in dog saphenous vein was due to experimental artifacts derived from the use of high concentration of EGTA and artificial pH-buffering reagent. PMID- 2557150 TI - Vascular receptors for angiotensin, vasopressin, and atrial natriuretic peptide in experimental hypertension. AB - Angiotensin II (ANG II) and vasopressin (AVP) are two powerful vasoconstrictors, and atrial natriuretic peptide (ANP) is a potent vasorelaxant. The changes in the density or affinity of binding sites for these agents that may alter target organ responsiveness in hypertension are reviewed. ANG II binding in mesenteric arteries was unaltered in one-kidney, one-clip (1-K, 1-C) and in 2-K, 1-C hypertensive rats, while in deoxycorticosterone acetate (DOCA)-salt hypertensive rats ANG II binding to blood vessels was significantly increased. A role of mineralocorticoids to increase the number of vascular ANG II sites in some hypertensive models is suggested. In spontaneously hypertensive rats (SHR) ANG II receptors were increased in young rats in the prehypertensive stage with respect to Wistar-Kyoto (WKY) control rats, but normal in older rats. AVP binding in the vasculature of hypertensive rats was uniformly decreased in inverse correlation to plasma AVP levels, but vascular responsiveness to AVP was exaggerated. Inositol trisphosphate production by blood vessels of SHR in response to AVP showed that increased AVP receptor-coupled phospholipase C activity may mediate in part the exaggerated pressor response in spite of reduced or normal density of receptors for vasoconstrictor peptides. Vascular ANP sites in 2-K, 1-C, 1-K,1-C, and DOCA-salt hypertensive rats varied inversely with plasma concentrations of ANP. Normal densities of ANP receptors in saralasin-sensitive 2-K, 1-C hypertensive rats correlated with ANP sensitivity, while saralasin-insensitive 2 K, 1-C hypertensive rats, which did not respond to ANP, had significantly decreased density of ANP vascular receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557151 TI - Chromosome 1 studies in Wilms' tumor. AB - Ten newly diagnosed patients with Wilms' tumor had blood and tumor samples taken for cytogenetic analysis. DNA was also extracted from these samples, along with blood obtained from both parents and an age- and sex-matched control. Molecular biological techniques were employed to study changes present in these samples with respect to chromosome 1. Two DNA probes, PIB 174 and PFBl, mapping to 1q12 qter and 1p12-pter, respectively, were examined for the presence of restriction fragment length polymorphisms (RFLPs) and to detect copy numbers of sequences homologous to the probes. These were normalized with respect to themselves and with regard to a control probe P30. No RFLPs were found with the restriction enzymes used. However, seven patients showed a marked alteration in hybridization signal in tumor and/or blood samples compared to control samples and the control probe. This was apparent using probe PFBl, but just failed to reach statistical significance using nonparametric testing. This would suggest that submicroscopic chromosome 1 changes are present more often in Wilms' tumor than previously recognized, and they may play a leading role in the genesis of this tumor. PMID- 2557152 TI - Beta-adrenergic receptors in congestive heart failure: present knowledge and future directions. AB - In recent years substantial information has become available on the function and regulation of beta-adrenergic receptors in experimental model systems and in the human heart. Beta-Adrenergic receptors mediate the positive inotropic and chronotropic effects of the sympathetic neurotransmitter norepinephrine in the heart. They can be altered in various disease states including congestive heart failure. In order to enhance understanding of beta-adrenergic receptor regulation in heart failure, we here review the present knowledge and the open question in three areas: (1) the differential role of beta 1- and beta 2-adrenergic receptors: (2) regulation of the number of cardiac beta-adrenergic receptors by drugs and disease states: and (3) regulation of the responsiveness of cardiac beta-adrenergic receptors. PMID- 2557154 TI - Perspectives on imaging modalities. AB - In the comparison of various techniques it is important to analyze what is known about the physical possibilities of a particular technique for making a specific measurement. X-ray CT, ultrasound, and emission methods with radionuclides do not measure chemical composition. NMR can uniquely measure chemical composition. Unfortunately in the measurement of chemical composition, NMR is insensitive, thus large volumes of tissue must be selected by the NMR spectroscopic experiment for a reliable measurement. Measurements that rely on tissue physical differences to detect surfaces and motion include rapid x-ray imaging and ultrasound. For these reasons they have some attributes for particular types of studies that are superior to emission tomography and NMR. However, in the past few years, improvements in the NMR technology have led to the ability to acquire images representing only 30 sec of time in 3-dimensional studies of the beating human heart. It was believed until recently that anatomical resolution of PET was so poor that other imaging modalities would have to be used to delineate the specific region from which the PET information was coming, particularly in studies of the brain. However, in the past year it has been demonstrated that PET can acquire high resolution images of the CNS cortical ribbon at 2.5 mm or better.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557153 TI - Transcatheter arterial chemoembolization with anticancer drug in iodized oil for primary hepatic carcinoma. AB - Transcatheter arterial chemoembolization using a suspension of anticancer drug in iodized oil combined with Gelfoam embolization was performed in 85 patients with advanced primary hepatic carcinoma. Long-term retention of iodized oil in the tumor was observed in 90% of the cases, and changes in tumor size were observable by abdominal X-ray film on follow-up studies. There was a decrease of tumor size in all cases and the overall 1-year survival rate was 43%. In tumors whose diameter was less than 20 cm and in patients without portal vein thrombosis the survival rate was nearly 60%. Combined treatment with anticancer drug in iodized oil and Gelfoam embolization of the proper hepatic artery was superior to use of the suspension alone. There was also better detection of small cancer nodules by the suspension as compared to conventional angiography. PMID- 2557156 TI - Possible involvement of platelet activating factor in anaphylaxis of passively sensitised, isolated guinea pig hearts. AB - There is evidence that cardiac tissue may be a target for antigen/antibody reactions. Platelet activating factor (PAF) is released during anaphylaxis and could mediate cardiac damage. To investigate this, guinea pigs were passively sensitised by anti-ovalbumin rabbit serum (6 mg.kg-1 intravenously) and 24 h later their hearts were excised and isolated according to a working heart preparation technique. After a 20 min equilibration period, anaphylactic challenge was induced by a bolus injection of ovalbumin (2 mg in 0.2 ml buffer) via the side arm of the aortic cannula. Heart rate, coronary flow, aortic flow, left ventricular developed pressure (LVDP), its first derivative (LVdp/dtmax) and left ventricular end diastolic pressure (LVEDP) were recorded. After ovalbumin challenge, heart rate and LVEDP were markedly increased, while coronary flow, aortic flow, LVDP, and LVdp/dtmax were profoundly decreased. All these alterations were over within 5 min, and the measured variables returned to approximately the pre-challenge values. BN 52021, a specific PAF receptor antagonist, was dissolved in the perfusion buffer and given in doses of 15, 30 and 60 mumol.litre-1 10 min prior to the induction of anaphylactic challenge until the end of the observation period. BN 52021 inhibited the increase in heart rate and LVEDP and the decrease in coronary and aortic flow, LVDP and LVdp/dtmax in a dose dependent manner. The changes produced by 30 and 60 mumol.litre-1 were statistically significant at the levels of p less than 0.01 and p less than 0.001 when compared to the control values.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557155 TI - Mexiletine-quinidine in isolated hearts: an interaction involving the sodium channel. AB - Combination therapy with mexiletine and quinidine has been shown to be more effective than either monotherapy in treating patients with ventricular tachycardia. This enhanced efficacy was associated with prolongation of ventricular refractoriness and conduction time in the infarct zone. As sodium channel activity is a determinant of both conduction time and refractoriness we formed the hypothesis that the mexiletine-quinidine interaction was due at least in part to interactions involving the sodium channel. To assess the role of sodium channel blockade in the enhanced anti-arrhythmic activity of mexiletine quinidine combination we determined whether the electrophysiological and anti arrhythmic effects of tetrodotoxin combined with mexiletine or quinidine mimicked the effect seen with mexiletine combined with quinidine. Eighty isolated perfused rabbit hearts were treated with mexiletine, quinidine and tetrodotoxin either alone or in combination before and after circumflex occlusion-reperfusion. Ventricular fibrillation occurred in response to single extrastimuli in all 24 hearts treated with a saline control infusion. Combinations of mexiletine and quinidine at concentrations which alone had little electrophysiological activity produced anti-arrhythmic activity greater than that seen with high concentrations of mexiletine or quinidine alone. The combination of similarly low concentrations of tetrodotoxin and quinidine also produced enhanced anti-arrhythmic efficacy and enhanced prolongation of ventricular refractoriness and conduction which mimicked the effect of mexiletine and quinidine in combination. In contrast, the combination of mexiletine and tetrodotoxin did not produce enhanced anti arrhythmic and electrophysiological activity. Since tetrodotoxin is a highly specific sodium channel blocker, these data suggest that the enhanced antiarrhythmic activity of mexiletine-quinidine combination therapy involves, at least in part, blockade of the cardiac sodium channel. PMID- 2557157 TI - Excision and insertion of the conjugative transposon Tn916 involves a novel recombination mechanism. AB - The covalently closed circular form of the conjugative transposon Tn916, which acts as an intermediate in transposition, is produced by a novel type of recombination. Excision of the element pairs noncomplementary base pairs, which flank the transposon in a heteroduplex, at the joint of a circular form. By a reversal of the excision process, the base pairs from the heteroduplex are inserted into the next target. We present a detailed molecular model for the movement of conjugative transposons that involves the initial formation of staggered nicks in the "coupling regions" that flank the inserted element. The different products of excision and insertion of Tn916 can be explained by this model. PMID- 2557158 TI - MOM19, an import receptor for mitochondrial precursor proteins. AB - We have identified a 19 kd protein of the mitochondrial outer membrane (MOM19). Monospecific IgG and Fab fragments directed against MOM19 inhibit import of precursor proteins destined for the various mitochondrial subcompartments, including porin, cytochrome c1, Fe/S protein, F0 ATPase subunit 9, and F1 ATPase subunit beta. Inhibition occurs at the level of high affinity binding of precursors to mitochondria. Consistent with previous functional studies that suggested the existence of distinct import sites for ADP/ATP carrier and cytochrome c, we find that import of those precursors is not inhibited. We conclude that MOM19 is identical to, or closely associated with, a specific mitochondrial import receptor. PMID- 2557159 TI - Herpes simplex virus induces a processing factor that stimulates poly(A) site usage. AB - Extracts from herpes simplex virus-infected cells and from mock-infected cells have been compared for their ability to process at RNA poly(A) sites in vitro. Nuclear extracts from infected cells contain an activity that increases processing efficiency specifically at a late herpes simplex virus poly(A) site. By contrast, a second virus poly(A) site is processed with equal efficiency by nuclear extracts from infected and mock-infected cells. Using precursor RNAs containing these two virus poly(A) sites in tandem, which allows ready detection of the processing factor, we show that this specific activity is heat labile. Analysis of RNAs produced by virus recombinants that contain the poly(A) site sequences in tandem also indicates that increased processing at the late virus poly(A) site occurs in vivo. PMID- 2557160 TI - The CaaX motif of lamin A functions in conjunction with the nuclear localization signal to target assembly to the nuclear envelope. AB - While the nuclear lamin proteins (A, B, and C) assemble specifically at the surface of the nuclear membrane, their sequences do not reveal stretches of hydrophobic amino acids that might explain their association with the nuclear membranes. However, the A and B lamin proteins possess Ras-like C-terminal CaaX sequence motifs, which in Ras proteins are sites of hydrophobic modifications required for membrane association and function. From the analysis of single and double lamin A mutants affecting the CaaX motif, the nuclear localization signal, and higher-order assembly properties, we propose that the CaaX motif functions as a nonspecific, low affinity membrane probe for proteins ultimately segregated to specific cellular membrane systems. Committed association with specific membranes requires additional interactions with membrane-resident factors. PMID- 2557161 TI - Molecular characterization of the two genes SNQ and SFA that confer hyperresistance to 4-nitroquinoline-N-oxide and formaldehyde in Saccharomyces cerevisiae. AB - The genes SNQ and SFA confer hyperresistance to 4-NQO and FA when present on a multi-copy plasmid in yeast. Both are non-essential genes since transplacement of SNQ by a disrupted snq-0::LEU2 yielded stable and viable haploid integrants. Southern analysis revealed that SNQ and SFA are single-loci genes, and OFAGE analysis showed that they are located on chromosome XIII and IV, respectively. Northern blot analysis of SNQ and SFA revealed poly(A)+ RNA transcripts of 2 kb and 1.7 kb, respectively. Nuclease S1 mapping showed SNQ to have a coding region of 1.6 kb and SFA, one of 1.3 kb. The 5' coding regions were determined for both genes, while the 3' end could only be determined for gene SNQ. Both genes do not appear to contain introns. The SFA locus was also mapped by transposon mutagenesis. Tn10-LUK integrants disrupted the SFA gene function at sites that were determined by subcloning to lie within the SFA transcription unit. PMID- 2557162 TI - The host range of the pKD1-derived plasmids in yeast. AB - pKD1 is a 2 mu-like circular plasmid found in the yeast Kluyveromyces drosophilarum that can also stably replicate in Kluyveromyces lactis. We have found a short intergenic region in this genome that appears to be functionally neutral; that is, the introduction of foreign sequences into the single EcoRI restriction site located near one of the inverted repeats did not affect the high stability of the natural plasmid. By introducing a G418 resistance gene at this site, we constructed an autonomous recombinant plasmid. Since this vector did not require cir+ hosts for its stable maintenance, it could be used to examine the transformation host range of pKD1 among all the species belonging to the genus Kluyveromyces. Both species closely related to K. drosophilarum as well as a few other species that are very different in chromosomal GC% could be transformed to yield highly stable transformant clones. PMID- 2557163 TI - Cloning and characterization of the SKI3 gene of Saccharomyces cerevisiae demonstrates allelism to SKI5. AB - We have identified a mutant strain of the yeast Saccharomyces cerevisiae which overproduces killer toxin. This strain contains a single mutation which fails to complement defects in both the SKI3 and SKI5 genes, while a cloned copy of this gene complements both the ski3 and ski5 defects. The level of secreted toxin from a cDNA based plasmid is not increased in a ski3 strain, showing that the overproduction phenotype is dependent upon an increased level of M1 dsRNA. PMID- 2557164 TI - Studies on proton pump inhibitors. II. Synthesis and antiulcer activity of 8-[(2 benzimidazolyl)sulfinylmethyl]-1,2,3,4-tetrahydroquinolines and related compounds. AB - Many 8-[(2-benzimidazolyl)sulfinylmethyl]-1,2,3,4-tetrahydroquinolin e derivatives were synthesized and tested for their (H+ + K+)adenosine triphosphatase (ATPase)-inhibitory and antisecretory activities against histamine induced gastric acid secretion in rats. These sulfinyl compounds were synthesized by the oxidation of the corresponding sulfides, which were obtained from the reaction of 8-chloromethyl-1,2,3,4-tetrahydroquinolines and 2 mercaptobenzimidazoles in the presence of potassium carbonate. All compounds tested were potent inhibitors of (H+ + K+)ATPase. Most of the compounds showed antisecretory activity. Among them, 8-[(2-benzimidazolyl)sulfinylmethyl]-1-ethyl 1,2,3,4-tetrahydro quinoline (IXa) was found to have the most potent activity. The structure-activity relationships are discussed. PMID- 2557165 TI - Facile synthesis of 8-benzoylthio-2,6-methano-3-benzazocines and 3 benzoylthiomorphinans having small-ring substituents. AB - Synthesis of 3-cyclopropylmethyl-, 3-cyclobutylmethyl-, and 3-methyl-8 benzoylthio-2,6-methano-3-benzazocines (1j-l) was performed by regio-selective chlorosulfonation of non-narcotic 8-deoxy derivatives (1a-c) followed by reduction and benzoylation. 3-Benzoylthiomorphinans (2h-j) were also obtained by the same method. Compounds having small-ring substituents (1k, 1l, 2i, 2j) were found to be weak but pure mu- and delta-opioid antagonists. The analgetic activity of 1k was almost equal to that of pentazocine. PMID- 2557166 TI - [Study on the effects of Dobells' solution]. PMID- 2557167 TI - [Acute bilateral glaucoma in an LAV-positive subject]. AB - A bilateral angle-closure glaucoma is described in a homosexual man, with a positive HIV serology. This angle closure is secondary to an anterior rotation of the ciliary body at the scleral spur following development of an inflammatory cilio-choroidal detachment: which is itself connected with a massive uveal retinal effusion. The cause of this uveal effusion appears to be cytomegalovirus. This unusual syndrome was the initial manifestation of AIDS (Acquired Immune Deficiency Syndrome). A symptomatologic treatment with cycloplegics and prednisolone and a specific one with DHPG (9-1, 3-dihydroxy-2-propoxymethyl guanine), resulted in rapid regression of the local lesions; his overall clinical condition however deteriorated rapidly over two months following onset of syndrome. PMID- 2557168 TI - Depletion of O6-alkylguanine-DNA alkyltransferase activity in mammalian tissues and human tumor xenografts in nude mice by treatment with O6-methylguanine. AB - We have previously shown that exposure of cells in culture to O6-methylguanine significantly reduces their level of the repair protein, O6-alkylguanine-DNA alkyltransferase (AGT), thus rendering cells more sensitive to the cytotoxic effects of chemotherapeutic chloroethylating agents. Experiments were carried out in mice to determine whether the AGT content of tissues and tumors could be reduced by in vivo treatment with O6-methylguanine. There was a dose-dependent decrease in AGT activity in liver tissues of CD-1 mice to 24% of basal levels after four hourly intraperitoneal injections of O6-methylguanine (110 mg/kg). Although the decline in AGT activity in the liver was reversible, the activity remained at 75% of basal levels for up to 25 h after the final injection. The effect of O6-methylguanine treatment on AGT activity was measured in mouse tissues as well as human colonic carcinoma tumors (HT29 and BE) grown in Swiss athymic nude mice. The activity in the liver, kidney, and spleen of these mice decreased to 33%-35% of control levels, whereas the activity in HT29 tumors was likewise diminished to 25% of control levels after four hourly injections of O6 methylguanine (100 mg/kg). There was no enhancement of the tumoricidal effectiveness of chloroethylating agents on the HT29 tumor after O6-methylguanine treatment, probably due to a disproportionately higher level of AGT in human tissue than in murine tissue. However, these studies suggest that O6 methylguanine can be given in vivo to examine the role of the AGT protein in protecting against the toxic and carcinogenic effects of alkylating agents. PMID- 2557169 TI - Prolongation of ifosfamide elimination half-life in obese patients due to altered drug distribution. AB - The pharmacokinetics of intravenous ifosfamide were determined in 16 patients with carcinoma of the bronchus. In all 25% (4) of these patients were obese (i.e. greater than 20% over their ideal body weight). The terminal elimination half life (t1/2 beta) was found to be higher in the obese group than in the control group (6.36 h, range 5.77-7.45 h) vs 4.95 h, range 1.82-6.48 h) (P less than 0.05). This prolongation of the elimination half-life was due to an increased volume of distribution (Vd beta) in the obese group (42.81 1, range 35.49-51.90 l) vs 33.70 l range (17.76-50.62 l) (P less than 0.05). There was therefore no significant difference in total plasma clearance between the obese and normal groups. No correlation of ifosfamide plasma half-life was observed with total body weight (TBW) or ideal body weight (IBW). However, a significant positive correlation was observed between the percentage of IBW and plasma half-life. A strong positive correlation was observed between IBW and the plasma clearance of ifosfamide. The Vd beta correlated with both TBW and the percentage of IBW, but not with IBW itself. When Vd beta was normalised for IBW, there was a strong positive correlation with the percentage of IBW, suggesting that ifosfamide distribution into the TBW is higher than that into the IBW. PMID- 2557170 TI - Inhibition of cellular esterases by the antitumour imidazotetrazines mitozolomide and temozolomide: demonstration by flow cytometry and conventional spectrofluorimetry. AB - Using flow cytometry and conventional spectrofluorimetry we have previously shown that chloroethylnitrosoureas (CNUs) can exhibit marked inhibition of cellular enzymes catalysing hydrolysis of fluorescein diacetate (FDA). More potent inhibition was seen for the carbamoylating CNUs, whereas alkylating agents were largely inactive. We now report results obtained with the developmental imidazotetrazines mitozolomide and temozolomide in comparison with BCNU, the novel alkylating agents clomesome and cyclodisone, and the active mitozolomide metabonate MCTIC. Inhibition of EMT6 mouse mammary-tumour esterases was seen for mitozolomide and temozolomide, and activity against purified porcine carboxylesterase was demonstrated. Flow cytometric analysis showed that inhibition occurred across the entire EMT6 cell population, with no evidence of a subpopulation resistant to enzyme inhibition. Inhibitory potency for the imidazotetrazines was much weaker than for BCNU. With EMT6 cells, I50 values from flow cytometry were 9.7 x 10(-3) M and 1.5 x 10(-3) M for mitozolomide and temozolomide compared with 3.7 x 10(-4) M for BCNU. These were higher than the ID50 values for in vitro antitumour activity (MTT assay), 8.5 x 10(-6) M in the case of mitozolomide and 1.2 x 10(-5) M for BCNU, but similar to that of 5.6 x 10(-4) M for the less toxic temozolomide. MCTIC and cyclodisone showed very low activity, but significant inhibition was seen for clomesome. The results are consistent with the view that the imidazotetrazines do not exhibit major carbamoylating ability, although significant effects are seen at cytotoxic concentrations of temozolomide. In addition, the potential for the generation of carbamoylating species at the enzyme active site cannot be ruled out. PMID- 2557171 TI - A phase II study of carboplatin and vincristine in previously treated patients with small-cell lung cancer. AB - A total of 28 previously treated patients with small-cell lung cancer were treated at relapse with 400 mg/m2 carboplatin and 2 mg vincristine on days 1 and 8, every 4 weeks. Ten partial responses (PRs) (37%) but no complete responses (CRs) were seen. Median survival after the start of second-line treatment was 120 days (range, 39-503 days). Toxicity of this regimen was moderate, including WHO grade 3/4 myelosuppression in 26% of courses (n = 66). Eight PRs were seen in a subgroup of 22 patients who relapsed less than 3 months after first-line treatment. The responses seen in this group of patients may be due to the absence of cross-resistance between the regimens used. PMID- 2557172 TI - Haematological recovery following high-dose cyclophosphamide with autologous bone marrow transplantation. AB - A total of 31 patients with previously untreated small-cell carcinoma of the lung were treated with very-high-dose cyclophosphamide, using autologous bone marrow transplantation (ABMT) to assist haematological recovery. The period of neutropenia was shorter with 40 mg/kg cyclophosphamide x 4 (7 patients) than when the dose of cyclophosphamide was increased to 50 mg/kg x 4 (11 patients), despite ABMT 2 days after chemotherapy in each group. In all, 13 patients were treated with 50 mg/kg cyclophosphamide x 4, with infusion of bone marrow delayed to day 4, 6 or 8 after chemotherapy to determine the contribution of ABMT to haematological recovery. The period of neutropaenia was increased when marrow was returned 6 days following chemotherapy, confirming that ABMT contributed to haematological recovery after this schedule of treatment. A total of 11 patients had a second cycle of 50 mg/kg cyclophosphamide x 4 after recovery from the first cycle of high-dose chemotherapy. The period of myelosuppression was greater with the second cycle of chemotherapy, although ABMT was carried out during both cycles. The results show that ABMT contributes to haematological recovery when the dose of cyclophosphamide is high enough to produce prolonged hypoplasia. The increased myelosuppression observed after a second high-dose treatment in spite of ABMT suggests either that both transplanted and endogenous marrow activity contribute to recovery of myelopoiesis or that there is residual damage to marrow stroma after the first cycle of treatment. The data indicate the necessity of carefully assessing the role of ABMT in haematological recovery with high-dose chemotherapy regimens. PMID- 2557173 TI - Effects of atrial natriuretic peptide on the coronary arterial vasculature in humans. AB - The effects of the synthetic 28-amino-acid alpha-human atrial natriuretic peptide (ANP) on the proximal coronary arteries and coronary blood flow were evaluated in 17 patients. Proximal coronary dimension was quantitated by digital angiography, and coronary flow was quantitated with 3F Doppler flow catheters. ANP, when given as a 2.5-micrograms/kg bolus in the left ventricle, caused sustained significant proximal coronary dilations from 3.49 +/- 0.57 to 4.09 +/- 0.76 mm, lasting more than 30 minutes. The proximal coronary diameter did not increase further after intracoronary injection of 0.3 mg nitroglycerin (4.08 +/- 0.79 mm). Coronary flow (resistance coronary dilation) was not significantly increased at 5 minutes after ANP (87 +/- 55 to 102 +/- 54 vol flow units), indicating that the proximal coronary dilations were not flow dependent. The persistent proximal coronary dilations were associated with minor and transient decreases in aortic pressure and left ventricular end-diastolic pressure and with minor and transient increases in heart rate, cardiac output, and left ventricular contractility. Plasma ANP level increased significantly by more than sixfold from 39.8 +/- 8.8 to 245.8 +/- 168.5 pg/ml. The time course of proximal coronary dilations was related more closely to the time course of increase in plasma cyclic guanosine monophosphate than that of plasma ANP. This study demonstrates that bolus injection of ANP (2.5 micrograms/kg), an endogenous vasodilator, caused marked sustained preferential proximal coronary dilations and brief minor changes in cardiac and systemic hemodynamics. Although additional studies are needed to assess its clinical efficacy as a coronary dilator in the treatment of coronary artery disease, these data suggest a potential of ANP in the therapy of ischemia. PMID- 2557174 TI - Alpha 2-receptor-mediated vasoconstriction in patients with congestive heart failure. AB - alpha 2-Adrenoceptors exist postsynaptically to subserve vasoconstriction and presynaptically to modulate norepinephrine release into the synaptic cleft. Because adrenoceptors may down-regulate in response to chronic stimulation, we investigated the activity of alpha 2-receptor-mediated vasoconstriction in patients with congestive heart failure, who had increased levels of plasma norepinephrine. We used the isolated forearm model and intra-arterial infusions of subsystemic doses of yohimbine, a specific alpha 2-blocker, in 11 patients with heart failure and in 15 normal subjects. Yohimbine produced a dose-related increase in forearm blood flow and decrease in forearm vascular resistance. These findings were consistent with a direct vasodilator effect mediated by blockade of the postsynaptic alpha 2-vascular receptor. Furthermore, the vasodilator responses in patients with heart failure were similar to the normal subjects in terms of the percent increase in forearm blood flow, the dose-response relation, and the fractional response to hyperemia and phentolamine; thus, alpha 2-receptor mediated vasoconstriction is neither enhanced nor down-regulated in heart failure. In addition, in patients with heart failure and in normal subjects, yohimbine produced an increase in the forearm venous norepinephrine concentration, consistent with an inhibition of the presynaptic alpha 2-receptor resulting in an augmented release of norepinephrine into the synaptic cleft. Thus, these data suggest that the postsynaptic alpha 2-receptor is an important mediator of vasoconstriction in patients with heart failure. Despite chronic elevations in plasma norepinephrine in patients with heart failure, alpha 2 receptor mechanisms subserving vasoconstriction and inhibition of norepinephrine release into the synaptic cleft are still functional in heart failure. PMID- 2557175 TI - Role of calcium on excitation-contraction coupling in cardiac and vascular smooth muscle. AB - The calcium ion is recognized as having a ubiquitous role in a wide range of physiological responses. The calcium entry blockers have assumed a greater role than first thought possible in the management of cardiovascular disorders. As a group, they have multiple effects and can be tailored to meet specific needs. The drugs are well tolerated, thus making them useful for chronic administration. The efficacy of the calcium entry blockers can be enhanced when combined with other therapies such as beta-adrenoceptor antagonists and inhibitors of angiotensin converting enzyme as applied to patients with hypertension, angina pectoris, or both. The range of indications and potential uses of the calcium entry blockers demands an understanding of the role of the slow inward current in cardiac and vascular smooth muscle. This review focuses on the pharmacological actions of the calcium entry blockers and relates these events to their clinical applications in an effort to achieve an understanding of their multiple therapeutic uses. PMID- 2557176 TI - Mechanisms of action and clinical use of calcium antagonists in hypertension. AB - Calcium antagonists are potent arterial vasodilators that do not lead to relevant chronic sympathetic reflex activation and sodium and volume retention. This favorable hemodynamic profile renders them suitable for monotherapy of hypertension in which they can reduce the calcium influx-dependent functional component of elevated vascular resistance that may be enhanced by altered vascular muscle cation handling and increased intracellular free calcium concentrations. Clinical studies have proved their efficacy, safety, and good tolerability alone or in combination with other drugs in uncomplicated hypertension in which they are particularly effective in older, low renin, and possibly, black patients. These properties and their efficacy in the treatment of severe and accelerated hypertension or hypertensive emergencies make them a valuable addition to already available drug therapy. PMID- 2557177 TI - Recent insights into the calcium channels. AB - Calcium channels play a central role in the regulation of intracellular calcium (Ca2+) concentration, and their function is subject to control by voltage regulated, receptor-regulated, or voltage- and receptor-regulated mechanisms. Three types of calcium channels have been described. These are the T (transient or "fast"), the N (neuronal), and the L (long lasting or "slow") channels. The L channels appear to be heterogeneous and have different properties in different tissues. Intracellular calcium-ion concentration can be increased by three types of receptor mechanisms. In the heart, L channels can be phosphorylated by a cyclic AMP-dependent protein kinase after beta 1-adrenergic receptor stimulation. In vascular smooth muscle, the postjunctional alpha 2-adrenergic receptor is coupled to a Ca2+ channel by a G protein; receptor stimulation facilitates calcium influx. This channel might be a form of L channel. A third receptor mechanism, especially active in vascular smooth muscle, is typified by the alpha 1-adrenergic receptor that, when stimulated, will activate phospholipase C. This leads to an increase in intracellular inositol trisphosphate (IP3), which is an intracellular messenger that can induce calcium release from the sarcoplasmic reticulum. Thus, release of norepinephrine from sympathetic nerves in the cardiovascular system stimulates the heart and vessels to contract by increasing Ca2+; however, the mechanism by which this occurs is different, depending on whether the noradrenergic agonist interacts with beta 1-, alpha 2-, or alpha 1 adrenergic receptors. PMID- 2557178 TI - Effects of calcium entry blockade on myocardial blood flow. AB - The calcium entry-blocking drugs produce effects on the coronary vasculature that might be expected to exert anti-ischemia activity. Although these agents cause little vasodilation of the epicardial coronary arteries during basal conditions, they block vasoconstriction that can increase stenosis severity during isometric exercise and interrupt coronary artery spasm in patients with variant angina. Administration of the calcium blockers causes transient vasodilation of the coronary resistance vessels, followed by decreased responsiveness to a brief ischemic stimulus. This results in decreased coronary reactive hyperemia after transient coronary occlusion. By preventing excessive ischemic vasodilation of the resistance vessels, these agents can enhance perfusion of the subendocardium distal to a flow-limiting coronary stenosis. The calcium entry blockers have relatively little effect on the immature coronary collateral vessels that exist at the time of acute coronary occlusion. Diltiazem, however, has been demonstrated to increase collateral blood flow in animals in which chronic coronary occlusion has resulted in growth of moderately well-developed collateral vessels. PMID- 2557179 TI - Resistance of metastatic pancreatic endocrine tumours after long-term treatment with the somatostatin analogue octreotide (SMS 201-995). AB - Ten patients with metastatic pancreatic endocrine tumours were treated with the long-acting somatostatin analogue octreotide (SMS 201-995). Three patients showed no response, clinically or biochemically, and treatment was therefore withdrawn. The seven remaining patients continued treatment for a median period of 28 months (range 13-54 months). Treatment was initially effective, symptoms improved and the concentrations of tumour-related hormones were reduced. Worsening of symptoms and rising levels of tumour-related hormone concentrations occurred a median of 5 months (range 1-6 months) after the start of therapy and were initially reversed by increasing the dose of octreotide over a median of 10 months (range 6-16 months). However, after a median of 13 months (range 5-34 months) at the maximum dosage, symptoms recurred and were no longer responsive to a further increase in dosage of octreotide or other therapeutic measures. All patients died within a period of 5 months once this resistant phase of their illness had been reached. PMID- 2557180 TI - Effects of the GABAergic agent sodium valproate on the arginine vasopressin responses to hypertonic stimulation and upright posture in man. AB - In order to evaluate the possible influence of GABAergic neurotransmission on the arginine vasopressin (AVP) response to osmotic and pressure volumetric stimuli, the GABAergic drug sodium valproate was administered by mouth (200 or 400 mg 16 h, 8 h and just before tests) to eight normal men before osmotic (i.v. infusion of 0.51 , NaCl for 2 h) and orthostatic (standing upright and maintaining an orthostatic position for 20 min) tests. In both experimental conditions, the AVP rise was significantly lower in the presence than in absence of sodium valproate. The maximum AVP responses in the control orthostatic and osmotic tests were respectively 2.3 and 2.5 times higher than basal levels. When 600 mg sodium valproate was given, the maximum AVP rise in response to hypovolaemic and osmotic stimuli were respectively 1.75 and 2.1 times higher than basal value. Similar results were obtained giving 1.2 g sodium valproate. These results suggest that in man a GABAergic pathway is involved in the AVP responses to hypovolaemic and hyperosmotic stimuli. PMID- 2557181 TI - [Subacute myeloneuropathy after abuse of nitrous oxide: an electron microscopic study on the peripheral nerve]. AB - We have experienced a case of myeloneuropathy following habitual abuse of nitrous oxide. We report clinical and pathological findings of this case with review of literatures. A 36-year-old dentist was first admitted to our hospital on August 17, 1983 because of numbness of both lower legs and unsteady gait. He had recreationally inhaled nitrous oxide 30 to 60 minutes everyday since a year ago. Neurological examination showed ataxic broad-based gait, moderate loss of pain and touch sensation in the lower limbs up to patella. Position and vibratory senses were more severely impaired. Deep tendon reflexes increased in the upper extremities and at the knees, but diminished at ankle jerks. Muscle strength was normal. Prominent Lhermitte's sign was present. Except for reduced serum vitamin B12 level, laboratory results were normal. Needle electromyography showed high amplitude and long duration NMU in the right quadriceps and anterior tibial muscles. Motor nerve conduction velocity was 34 m/sec at the right posterior tibial nerve and could not be detected at the left. Sural nerve biopsy was performed. The density of the myelinated fibre measured on transverse section was in normal range but degenerated fibres were occasionally recognized. Single teased nerve fibre showed various degree of myelin ovoid along the fibre. Myelin loss was shown to occur in some parts of the fibre. Electron microscopy showed myelin splitting and formation of intramyelinic vacuoles containing myelin debris. Axon was almost normal at least in the early stage of degeneration. Later, axon disappeared with destruction of myelin sheath. These nerve changes largely demonstrated demyelination but it was occasionally accompanied with axonal degeneration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557182 TI - [Two cases of mitochondrial myopathy (focal cytochrome c oxidase deficiency), long-term follow-up on a diagnosis of ocular type myasthenia gravis]. AB - Two patients (a 50-year-old and a 35-year-old men) with focal cytochrome c oxidase deficiency, manifesting ptosis and external ophthalmoplegia of 13 and 6 years' duration, respectively, were reported. Patient 1 (a 50-year-old male) had also slight muscular weakness of the proximal limb and neck flexor muscles. Diagnosis of myasthenia gravis had been made on the clinical findings including ptosis and external ophthalmoplegia, diurnal fluctuation of symptoms, and equivocal positive Tensilon test. However, waning phenomenon on repetitive nerve stimulation or elevation of titer of the anti-acetylcholine receptor antibody was not detected on both patients. Needle EMG showed mild myopathic changes. Finally, pathological and biochemical analyses of the biopsied muscles confirmed the diagnosis of mitochondrial myopathy (focal cytochrome c oxidase deficiency). PMID- 2557183 TI - The effects of gastric pH and food on the pharmacokinetics of a new oral cephalosporin, cefpodoxime proxetil. AB - The effects of alteration of gastric pH and food on the pharmacokinetics of 200 mg doses of cefpodoxime proxetil tablets were studied in two separate randomized, open label, crossover studies in healthy subjects. In the pH study (n = 17 subjects), there was a lead-in period done under fasting conditions, followed by randomization to a four-way crossover of pentagastrin (6 micrograms/kg, subcutaneously), ranitidine (150 mg orally, 10 and 2 hours before dosing with the antibiotic), sodium bicarbonate (12.6 gm), or aluminum hydroxide (120 cc). Gastric pH was determined by nasogastric aspirates before and 10 minutes after the intervention, just before the antibiotic was given. Peak plasma concentrations (Cmax) and area under plasma concentration-time curve (AUC) were highest in fasting and pentagastrin periods and were 35% to 50% lower for all of the other periods (p less than 0.0001). Gastric pH and Cmax and AUC were inversely related (r = 0.66 and r = 0.62; p less than 0.0001 for both). In the food study (n = 16 subjects), there were two lead-in periods, one done while subjects were fasting and one while they were normal diet, followed by randomization to a four-way crossover of either high or low protein diets, or high or low fat diets. There were six meals in each diet. Dosing with the antibiotic was done at the midpoint of the fourth meal. Cmax and AUC were 22% to 34% higher for all diets than for the fasting period (p less than 0.0001), whereas the time to Cmax was unchanged. These studies demonstrated that absorption of cefpodoxime proxetil is best at low gastric pH or in the presence of food, which suggests that the role of gastrointestinal function on the pharmacokinetic profile is complex. PMID- 2557185 TI - Effects of L-1-methyl-histidine and the muscle dipeptides carnosine and anserine on the activities of muscle calpains. AB - 1. Carnosine, anserine and L-1-methyl-histidine activated muscle calpain II assayed at 2.5 mM Ca2+. 2. At 5 microM Ca2+, none of these compounds activated calpain II sufficiently to bring its activity up to the level measured at 2.5 mM Ca2+. 3. Carnosine increased, whereas both anserine and L-1-methyl-histidine decreased the inhibitory effect of calpastatin on calpain II. 4. These results suggest that although the compounds are not potent activators of calpain II, the ratio of the dipeptides in muscle may have an effect on calpain II-calpastatin interaction. PMID- 2557184 TI - The relative clinical effectiveness of three anticalculus dentifrices. AB - A three-month double-blind clinical study was conducted to compare the effect of three commercially available antitartar dentifrices in retarding the rate of supragingival calculus formation. After a thorough dental prophylaxis, the subjects were instructed to brush their teeth twice daily with either Prevent, Crest Tartar Control Formula (TCF), or Colgate TCF. Supragingival calculus accumulation on the mandibular anterior teeth was measured after 3 months use of the assigned dentifrice. The results of this study showed that Prevent retarded calculus formation 38.0% more than Crest TCF. Colgate TCF retarded calculus accumulation by 36.0% more than Crest TCF. No significant differences in anticalculus effectiveness were noted between Prevent and Colgate TCF. PMID- 2557186 TI - Expression of hemolytically active human complement component C1r proenzyme in insect cells using a baculovirus vector. AB - The gene of human C1r has been expressed in a baculovirus-insect-cell system via the pAc373 transplacement vector. The full-length cDNA copy was inserted into the pAc373 vector downstream from the strong polyhedrin promoter of the baculovirus, Autographa californica nuclear polyhedrosis virus (AcNPV). Spodoptera frugiperda cells were cotransfected with the resultant plasmid, pAcC1r, and the wild-type AcNPV DNA. Recombinant viruses, which drove the expression of C1r protein, were selected by plaque morphology and ELISA. Insect cells infected with the recombinant virus produced and secreted human C1r protein, at a level of 1-2 mg/l of medium. The expressed C1r was isolated from the medium by chromatofocusing. On reducing gels only a single Coomassie-staining band was observed, and this band migrated at 80-83 kD characteristic of the unactivated C1r proenzyme. Its identification as C1r was immunologically confirmed on Western blots. C1 reconstituted from purified C1r expressed in insect cells together with human C1q and C1s proved biologically active in a hemolytic assay. Thus, the baculovirus insect-cell system is capable of expressing and secreting a sophisticated, multifunctional human complement subcomponent in its biologically activatable form. PMID- 2557187 TI - Renal handling and metabolism of adenosine in diabetic rats. PMID- 2557189 TI - Subperiosteal implants fill niche when ridge is too thin. PMID- 2557188 TI - [Maxillo-mandibular rehabilitation and implantology (bone and biomaterials)]. PMID- 2557190 TI - [Cyclic AMP levels in the plasma and ischemic myocardium and free fatty-acid levels in the serum following coronary artery ligation in dogs: relation to ventricular fibrillation]. AB - This study was designed to shed light on the biochemical causes of serious ventricular arrhythmias in the early stage of acute myocardial infarction (AMI). Fourteen mongrel dogs were divided into 2 groups. One group was subjected to coronary ligation by placing silk sutures around the LAD coronary artery about 1.5 cm from its origin. All the dogs of this group were found to have ventricular tachycardia and fibrillation, starting approximately 18 min. After ligation, associated with regional accumulation of myocardial cAMP in the ischaemic zone. The increase of cAMP started about 10-15 min. before the onset of the arrhythmia. In contrast, the control group had only sutures placed without ligation and was not found to have change in myocardial cAMP within 35 min and arrhythmias at the same time. Both plasma cAMP and serum FFA were studied, but no significant changes were found. It is concluded that myocardial cAMP may play an important role in the genesis of ventricular arrhythmias in the acute ischaemic heart and it may be one of the arrhythmogenic factors to evoke ventricular arrhythmias. PMID- 2557191 TI - Effect of adenosine and adenosine analogs on [14C]aminopyrine accumulation by rabbit parietal cells. AB - Adenosine receptors that modulate adenylate cyclase activity have been identified recently in a number of tissues. Adenosine A2 receptor is stimulatory to adenylate cyclase, whereas adenosine A1 receptor is inhibitory to adenylate cyclase. We investigated the effect of adenosine and its analogs on [14C]aminopyrine accumulation by rabbit parietal cells. Rabbit gastric mucosal cells were isolated by enzyme digestion. Parietal cells were enriched by nonlinear percoll gradients. [14C]Aminopyrine accumulation was used as an indicator of acid secretion. The effect of 2-chloroadenosine on histamine stimulated [14C]aminopyrine accumulation was studied. The effects of N ethylcarboxamideadenosine, 2-chloroadenosine, stable analogs of adenosine, and adenosine on [14C]aminopyrine accumulation were assessed. Cyclic AMP content of parietal cells was determined by radioimmunoassay. Histamine and carbachol, known secretagogues, stimulated [14C]aminopyrine accumulation. 2-Chloroadenosine did not suppress histamine-stimulated [14C]aminopyrine accumulation. 2 Chloroadenosine, N-ethylcarboxamideadenosine, and adenosine dose dependently increased [14C]aminopyrine accumulation. The order of potency was N ethylcarboxamideadenosine greater than 2-chloroadenosine greater than adenosine. 8-Phenyltheophylline and theophylline, adenosine-receptor antagonists, or cimetidine did not have significant effects on the increase of AP uptake induced by 2-chloroadenosine. Coadministration of dipyridamole, and adenosine uptake inhibitor, augmented the effect of adenosine on [14C]aminopyrine accumulation. 2 Chloroadenosine, N-ethylcarboxamideadenosine, and adenosine each induced a significant increase in cellular cyclic AMP. We conclude that there may be adenosine A2 receptors on rabbit parietal cells which modulate gastric acid secretion. PMID- 2557192 TI - Autonomic function and motility in intestinal pseudoobstruction caused by paraneoplastic syndrome. AB - This report documents the occurrence of chronic intestinal pseudoobstruction in association with a small cell carcinoma of the lung with evidence of pre- and postganglionic sympathetic dysfunction in one patient with brain metastases, and with sympathetic and parasympathetic postganglionic dysfunction in a second patient. A strategy is outlined for the identification and characterization of disordered neural control of gut motility. This strategy utilizes gastrointestinal motility studies to confirm gut neuropathy, autonomic function tests, and plasma norepinephrine responses to intravenous edrophonium to identify the level of dysfunction. These cases are compared with others in the literature, and the occult nature of these cancers, the spectrum of symptoms suggesting autonomic dysfunction on presentation, and the occasional response of the neurologic deficit to treatment of the malignancy are highlighted. PMID- 2557193 TI - Octylonium bromide interacts competitively with the PAF receptor. AB - To clarify the nature of the interaction of octylonium bromide with the PAF receptor, saturation studies of 3H-PAF binding were made in the presence of increasing concentrations of the radioligand (from 0.06 to 7.4 nM) and of octylonium bromide (1 X 10(-8), 5 X 10(-8) and 1 X 10(-7) M) or of a potent PAF antagonist L 652,731 (2.5 X 10(-7) M), or in the absence of competing drugs. Saturation binding data were plotted according to the non-linear fitting analysis and to Scatchard transformation. Both mathematical models indicated that the presence of added drugs decreased the affinity of the complex 3H-PAF/PAF receptors, leaving unaffected the maximum number of PAF binding sites. Receptor parameters in the absence of competitors were: KD = 1.2 nM, Bmax = 950 fmoles 3H PAF bound/10(8) platelets. Octylonium bromide increased, in a concentration dependent manner, KD values from 1.5 to 2.1 nM, while Bmax ranged from 910 to 980 fmoles 3H-PAF specifically bound/10(8) platelets. Likewise, L 652,731 did not influence Bmax (980 fmoles) but increased KD (1.9 nM). The binding behaviour of octylonium bromide and L 652,731 indicates that the two compounds inhibit competitively the binding of 3H-PAF to its receptors. PMID- 2557194 TI - [Direct detection of oxygen free radicals produced by myocardial reperfusion using electron spin resonance spectroscopy]. AB - There is a growing evidence for the role of oxygen free radicals (OFR) in mediating myocardial tissue injury during myocardial ischemia and particularly during reperfusion. But almost all of the evidence was indirect, using electron spin resonance (ESR) spectroscopy, we have directly measured OFR generated in ischemic and reperfused isolated rabbit hearts. 17 hearts were rapidly frozen in liquid nitrogen after their arrest by cardioplegic solution and sampled after 150 min of sustained hypothermic global ischemia or after reperfusion. The ESR spectra obtained from experiment have directly demonstrated that OFR is produced in significant amounts in the isolated rabbit hearts during early stage of reperfusion but only small amount during ischemia. The mitochondrial electron transport chain appeared to be the main source of OFR. We found that superoxide dismutase scavenged OFR generated during reperfusion efficiently, but catalase did not. We believe that superoxide anion, not hydroxyl radical, is the main OFR which is responsible for myocardial reperfusion injury. We also found that Salvia, a traditional Chinese medicine, a very efficient OFR scavenger, had the similar effect as superoxide dismutase. PMID- 2557195 TI - [Control measures in officially acknowledged brucellosis-free and leukosis unsuspected dairy herds on the basis of bulk milk samples in combination with ELISA tests]. AB - 1. EC- and National Regulations. Since 1988 the EC-regulations accept in addition to the on Agar Gel Immunodiffusion test (AGIDT) based blood serum testing of cattle herds that are filed as "free from Enzootic Bovine Leucosis" the use of ELISA for this purpose. The regular testings in dairy cattle herds can be done alternatively with single or pooled milk samples, in other herds with pooled blood sera using ELISA. General condition is only a minimal sensitivity of the test to detect the European EBL Antibody Standard ("E4") in a dilution of 1:10 in negative serum or 1:250 in negative milk. Adequate national regulations are in preparation. The present limitation of pool sizes, blood maximum 50 animals without preparation steps 20, and milk after concentration treatment 50 cows is neutralized by proceedings in development of higher sensitive ELISA tests. This limitation should be canceled. Herd bulk milk samples without size limitations are accepted to be tested with "Milk Ring Test" by EC for the regular testings in filed "Brucellosis Free Dairy Cattle Herds". The alternative use of more sensitive (and more specific) ELISA tests for this purpose including the technical conditions is in a final discussion. 2. Scientific-Technical Base for Using the Chances of the Proceeding in the EC-Regulations. The realisation of the EC accepted or final discussed ELISA based bulk milk testing to control filed "EBL- and/or Brucellosis Free Herds" depends on some basic conditions like sensitivity, specificity, and variability of the ELISA systems. Field trials of more than 20,000 bulk milk samples in case of Brucellosis and more than 2,000 in case of EBL show the feasibilities and the limits of the ELISA systems in defining the status of the herds. The Brucellosis respectively the EBL situations of the dairy cattle herds tested in this trail were well known by history and by investigation of single animal blood samples using conventional tests. Special test run variations of pretested assays demonstrated the possibilities to define the EBL status of dairy cattle herds up to 50 lactating cows without preparation of the bulk milk sample and up 100 after concentration of the antibodies by the rennet-ammonium sulfate method. The concentration limit for detection of Brucellosis antibodies is 100 lactating cows. The bulk milk of smaller herds can be tested without concentration. On principle the evaluation of the test values bases on defined relations to a "weak positive" reference.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557196 TI - Inducible expression of an hsp68-lacZ hybrid gene in transgenic mice. AB - Transgenic mice have been generated that express the E. coli beta-galactosidase gene under the control of the promoter from the mouse heat-shock gene, hsp68. Sequences from -664 to +113 relative to the start of transcription of the hsp68 gene were sufficient to direct stress-induced expression of the beta galactosidase gene in adult tail tissue and various tissues of fetal stages of development. Expression was detected in situ by staining with the chromogenic substrate, X-gal. The hybrid gene was refractory to induction in preimplantation embryos until the blastocyst stage of development, as reported for the endogenous hsp68 gene. No constitutive expression was observed by in situ staining or Northern analysis at any stage of development, even in tissues that constitutively express the endogenous hsp68 gene. We conclude that the hsp68 promoter region included in the construct contains sufficient sequence information for heat and arsenite inducibility, but it does not contain sequences controlling tissue-specific expression during development. This tightly regulated inducible promoter may provide a useful tool for short-term inducible gene expression in transgenic mice. PMID- 2557197 TI - Cyclic AMP waves during aggregation of Dictyostelium amoebae. AB - During the aggregation phase of their life cycle, Dictyostelium discoideum amoebae communicate with each other by traveling waves of cyclic AMP. These waves are generated by an interplay between random diffusion of cyclic AMP in the extracellular milieu and the signal-reception/signal/relaying capabilities of individual amoebae. Kinetic properties of the enzymes, transport proteins and cell-surface receptor proteins involved in the cyclic AMP signaling system have been painstakingly worked out over the past fifteen years in many laboratories. Recently Martiel & Goldbeter (1987) incorporated this biochemical information into a unified mathematical model of communication among Dictyostelium amoebae. Numerical simulations of the mathematical model, carried out by Tyson et al. (1989), agree in quantitative detail with experimental observations of cyclic AMP traveling waves in Dictyostelium cultures. Such mathematical modeling and numerical experimentation provide a necessary link between detailed studies of the molecular control mechanism and experimental observations of the intact developmental system. PMID- 2557198 TI - Lithium changes the ectodermal fate of individual frog blastomeres because it causes ectopic neural plate formation. AB - Amphibian blastulae that are treated with lithium (Li) develop into embryos that consist almost exclusively of head structures. This dramatic change in embryogenesis may occur either because Li selectively kills trunk progenitors or because Li causes trunk progenitors to become head progenitors. To distinguish between these possibilities, we compared the fates of individual frog blastomeres between Li-treated embryos and normal embryos using lineage tracers. The results demonstrate that Li causes ventral midline cells, which normally populate large amounts of trunk, to produce many head structures, including the brain. Examination of fluorescently labeled clones in living Li-treated gastrulae shows that: (1) the ectodermal members of the clones migrate normally, and chordamesodermal involution begins normally; (2) the chordamesoderm's later involution is altered such that it is confined to the vegetal hemisphere; (3) accordingly, the neural plate forms in the vegetal hemisphere, circumscribing the blastopore, which normally gives rise to the cloaca; and (4) the ectodermal progeny of the ventral midline blastomeres that are near the blastopore populate the brain because they are induced by the stalled chordamesoderm to form part of the ectopic neural plate. These results demonstrate that Li, administered during a short developmental window at early cleavage stages, ultimately alters ectodermal fate because it changes the pattern of chordamesodermal involution during gastrulation, which in turn changes the site of neural plate formation. PMID- 2557199 TI - [Augmentation of severely atrophied jaws with hydroxyapatite from a prosthodontics standpoint]. AB - After 4 years of routine application of hydroxylapatite ridge augmentation 109 patients underwent a posttreatment examination. The quality of the alveolar ridge and the denture as well as the functional status of the craniomandibular system were evaluated in detail. For the maxilla good results were obtained in 80-90%, for the mandible in 70-80%. Further improvements seem more probable by the avoidance of prosthodontic faults and the elimination of functional disturbances than by additional surgical procedures such as vestibuloplasty. PMID- 2557200 TI - [Reconstruction of atrophied jaw segments using individually configured hydroxyapatite vicryl mesh grafts. Experimental animal studies and clinical case reports]. AB - Individually configurated vicryl mesh sleeves permit the production of unusual implant shapes for the reconstruction of atrophied jaw segments. The experimental animal studies have shown that the vicryl mesh sleeve does not interfer with the bony integration of the implanted HA ceramics. The formation of a ceramoosseous regenerate is limited to the vicinity of the bony implant bed. PMID- 2557201 TI - [Comparative test of the soluble toxicity of hydroxyapatite ceramics using human and animal osteoblasts]. AB - The soluble toxicity of 5 different hydroxylapatite granulates was tested in osteoblast cell cultures. We established a human osteoblast-like cell culture and a culture of rat (Lewis) osteoblast-like cells. The granulates were also tested in cultures of human gingival fibroblasts as a conventional system. It is demonstrated that the osteoblast-like cell cultures are of higher sensitivity than fibroblast cultures which do not show any reaction to the extracts of hydroxylapatite granulates. Only 2 hydroxylapatite granulates show some toxic. The human osteoblast-like cells reacted slightly more sensitive to toxic substances of the materials tested than rat osteoblast-like cells. It is demonstrated that the osteoblast-like cell cultures are a highly sensitive test system for materials with low toxicity. PMID- 2557202 TI - Electroneuromyographic sequelae of tetanus, a controlled study of 40 patients. AB - To assess the peripheral nervous system involvement caused by tetanus 40 patients recovered from tetanus and their sex and age matched referents were submitted to electroneuromyography (ENMG). Clinical symptoms and signs of peripheral neuropathy were found in 6 patients, 3 of whom had other obvious aetiology than tetanus. EMG was examined of the masseters and three muscles in the extremities. Motor conduction velocities (MCVs) were examined of the median, ulnar and peroneal nerves, distal sensory conduction velocity (dSCV) of the median, ulnar and radial nerves and the sensory conduction velocity (SCV) of the median nerve. Repetitive stimulation was done to study the neuromuscular transmission. The patients had significantly longer motor distal latencies (DLs) and slower dSCVs and more frequent ENMG findings compatible with mono- or polyneuropathy than their referents. There was no systematical difference in the MCVs. These findings suggest that tetanus may cause toxic axonal polyneuropathy, sequels of which appear to be slight or subclinical and can be seen at a group level. PMID- 2557203 TI - An epidemiological and an experimental study on the effect of olive oil on total serum and HDL cholesterol in healthy volunteers. AB - We have studied the effects of olive oil on serum lipids in a dietary experiment with 48 healthy adult Dutch volunteers and in 76 boys 8-10 years of age from rural Crete. The dietary trial was carried out with normolipidaemic subjects, and it compared the effects of an olive-oil-rich and a carbohydrate-rich, high-fibre diet on serum lipids. Replacement of 12% of energy (en%) from saturated fatty acids by either monounsaturates or complex carbohydrates caused the same fall in serum total cholesterol of 0.45 mmol 1(-1). HDL cholesterol levels fell by 0.19 mmol 1(-1) on the carbohydrate-rich diet, but remained virtually unchanged on the olive-oil-rich diet. This was in agreement with our hypothesis that the total amount of fat in the diet is a determinant of HDL cholesterol. The diet of the Cretan boys contained 10 en% saturated fat, but 27 en% monounsaturated fat due to the liberal use of olive oil. Serum-lipid levels of the Cretan boys were not different from those of their counterparts from western European Countries. The mean body mass index of the Cretan boys was about 2 kg m-2 higher than those of boys in the other countries, which might have confounded comparisons. Whatever the explanation, the present-day lifestyle and diet in Crete do not produce a particularly favourable HDL to LDL cholesterol ratio in the blood, even though olive oil intake is high. PMID- 2557204 TI - Mediterranean diet and cancer. AB - A number of cancers, such as cancer of the large bowel, breast and other hormonal dependent organs, are less frequent in Mediterranean countries than in northern Europe. It has been hypothesized that a low dietary intake of saturated fat, accompanied by a higher intake of unrefined carbohydrates, and possibly other protective nutrients could be the cause of such risk differences. Cohort and case control studies based on individual dietary intake, however, cannot definitely corroborate the theory, nor disprove it. Analytical studies on fat, fibre and breast and colon cancers are reviewed and priorities for further studies are discussed. PMID- 2557205 TI - Real-time continuous-flow spin trapping of hydroxyl free radical in the ischemic and post-ischemic myocardium. AB - Real-time monitoring of spin-trapped oxygen-derived free radicals released by the isolated ischemic and reperfused rat heart has been achieved by ESR analysis of the coronary effluents using continuous flow detection and high-speed acquisition techniques. Two nitrone spin traps 5,5-dimethyl pyrroline 1-oxide (Me2PnO) and 3,3,5,5-tetramethyl pyrroline 1-oxide (MePnO) have been separately perfused at a concentration of 40 mM during a sequence of 50 min of low-flow ischemia (1 ml/min) followed by 30 min of global ischemia and subsequent reperfusion at the control flow rate (14 ml/min). ESR spectra were sequentially obtained in 5-min or 30-s blocks during low-flow ischemia and reperfusion, respectively. 1. The results show the formation of OH. free radicals in the ischemic and reperfused heart, as demonstrated by the observation of Me2PnO-OH (aN = aH = 14.9 G; g = 2.0053) and Me4PnO-OH (aN = 15.2 G, aH = 16.8 G; g = 2.0055) spin adducts. There is no evidence of significant biological carbon-centered or peroxyl free radicals spin-adduct formation in the coronary effluents or in lipid extracts analyzed after reflow. 2. The OH. generation began 15-20 min after the onset of ischemia and was moderate, peaking at 30-40 min. During reperfusion, an intense formation of OH. spin adducts was observed, with a maximum at 30-60 s and a further gradual decrease over the following 2 min. 3. Cumulative integrated values of the amount of spin adducts released during the ischemic period show a Me2PnO-OH level fourfold greater than that of Me4PnO-OH. It was 2.5 times greater during reflow, reflecting slower kinetics with the more stable Me4PnO. 4. The original ESR detection technique developed in this study allows accurate real-time quantitative monitoring of the oxygen-derived free radicals generated during myocardial injury. It might provide a quick and reliable new means for assessing the efficacy of free-radical inhibitors. PMID- 2557206 TI - ATP synthesis coupled to methane formation from methyl-CoM and H2 catalyzed by vesicles of the methanogenic bacterial strain Go1. AB - Methanogenesis from methyl-CoM and H2, as catalyzed by inside-out vesicle preparations of the methanogenenic bacterium strain Go1, was associated with ATP synthesis. That this ATP synthesis proceeded via an uncoupler-sensitive transmembrane proton gradient was concluded from the following results: 1. Various inhibitors that affected methane formation (e.g. 2-bromomethanesulfonate) also prevented ATP synthesis. 2. The protonophore 3,5-di-tert-butyl-4 hydroxybenzylidenemalononitrile, in combination with the K+ ionophore valinomycin, inhibited ATP synthesis completely without affecting methanogenesis. 3. The ATP synthase inhibitor diethylstilbestrol inhibited ATP synthesis. 4. Addition of the detergent sulfobetaine inhibited both methane formation and ATP synthesis; the former but not the latter could be restored by adding titanium(III) citrate as electron donor. In addition it was shown that ATP synthesis could also be driven by transmembrane proton gradients artificially imposed on the vesicles. Furthermore net methanogenesis-dependent ATP formation was shown by measuring [32P]phosphate incorporation. PMID- 2557207 TI - Phosphagen kinase evolution. Expression in echinoderms. AB - Arginine kinase and creatine kinase that catalyze the transfer of a phosphate group between ATP and arginine and creatine, respectively, play an important role in cellular energetics. In contrast to most animals which exhibit a single phosphagen kinase activity (creatine kinase in chordates and arginine kinase in protostomians), echinoderms exhibit both arginine kinase and creatine kinase activities, sometimes in the same tissue. In contrast to chordates in which creatine kinases are dimers (consisting of two subunits of 40 kDa) and protostomians in which arginine kinases are usually monomers (40 kDa), echinoids contain specific phosphagen kinases: a dimeric arginine kinase (consisting of two subunits of 42 kDa) in eggs and a monomeric creatine kinase (145 kDa) in sperm. We have examined echinoderms from the five existing classes (echinoids, asteroids, ophiuroids, holothurians and crinoids) for the expression of these specific phosphagen kinases in different tissues. Gel filtration was used to determine the molecular masses of the native enzymes. Antibodies specific for arginine kinase or for creatine kinase were used to characterize the subunit composition of arginine kinase and creatine kinase after SDS/PAGE and transfer. In all echinoderms analyzed, arginine kinase always occurred as an enzyme of about 81 kDa consisting of two subunits of 42 kDa and creatine kinase as a monomeric enzyme of 140-155 kDa. The occurrence in echinoderms of both phosphagen kinases with distinct specificities and specific molecular structures is discussed from both a developmental and evolutionary point of view. PMID- 2557208 TI - The effect of caldesmon on dynamic properties of F-actin alone and bound to heavy meromyosin and/or tropomyosin. AB - The effect of caldesmon on the rotational dynamics of actin filaments alone or conjugated with heavy meromyosin and/or tropomyosin has been measured by the electron paramagnetic resonance (EPR) technique using a maleimide spin label rigidly bound to Cys374 of actin. The rotation of actin protomers in filaments and the angular distribution of spin probes on actin were determined by conventional EPR spectroscopy, while torsional motions within actin filaments were detected by saturation transfer EPR measurements. Binding of caldesmon to F actin resulted in the reduction of torsional mobility of actin filaments. The maximum effect was produced at a ratio of about one molecule of caldesmon/seven actin protomers. Smooth muscle tropomyosin enhanced the effect of caldesmon, i.e. caused further slowing down of internal motions within actin filaments. Caldesmon increased the degree of order of spin labels on F-actin in macroscopically oriented pellets in the presence of tropomyosin but not in its absence. Computer analysis of the spectra revealed that caldesmon alone slightly changed the orientation of spin probes relative to the long axis of the filament. In the presence of tropomyosin this effect of caldesmon was potentiated and then approximately every twentieth protomer along the actin filament was affected. Caldesmon weakened the effect of heavy meromyosin both on the polarity of environment of the spin label attached to F-actin and on the degree of order of labels on actin in macroscopically oriented pellets. Whereas the former effect of caldesmon was independent of tropomyosin, the latter one was observed only in the absence of tropomyosin. PMID- 2557209 TI - Specificity of Bacillus thuringiensis delta-endotoxins. Importance of specific receptors on the brush border membrane of the mid-gut of target insects. AB - To study the molecular basis of differences in the insecticidal spectrum of Bacillus thuringienesis delta-endotoxins, we have performed binding studies with three delta-endotoxins on membrane preparations from larval insect mid-gut. Conditions for a standard binding assay were established through a detailed study of the binding of 125I-labeled Bt2 toxin, a recombinant B. thuringiensis delta endotoxin, to brush border membrane vesicles of Manduca sexta. The toxins tested (Bt2, Bt3 and Bt73 toxins) are about equally toxic to M. sexta but differ in their toxicity against Heliothis virescens. Equilibrium binding studies revealed saturable, high-affinity binding sites on brush border membrane vesicles of M. sexta and H. virescens. While the affinity of the three toxins was not significantly different on H. virescens vesicles, marked differences in binding site concentration were measured which reflected the differences in in vivo toxicity. Competition experiments revealed heterogeneity in binding sites. For H. virescens, a three-site model was proposed. In M. sexta, one population of binding sites is shared by all three toxins, while another is only recognized by Bt3 toxin. Several other toxins, non-toxic or much less toxic to M. sexta than Bt2 toxin, did not or only marginally displace binding of 125I-labeled Bt2 toxin in this insect. No saturable binding of this toxin was observed to membrane preparations from tissues of several non-susceptible organisms. Together, these data provide new evidence that binding to a specific receptor on the membrane of gut epithelial cells is an important determinant with respect to differences in insecticidal spectrum of B. thuringiensis insecticidal crystal proteins. PMID- 2557210 TI - The sodium cycle in methanogenesis. CO2 reduction to the formaldehyde level in methanogenic bacteria is driven by a primary electrochemical potential of Na+ generated by formaldehyde reduction to CH4. AB - CH4 formation from CO2 and H2 rather than from formaldehyde and H2 in methanogenic bacteria is inhibited by uncouplers, indicating that CO2 reduction to the formaldehyde level is energy-driven. We report here that in Methanosarcina barkeri the driving force is a primary electrochemical sodium potential (delta mu Na+) generated by formaldehyde reduction to CH4. This is concluded from the following findings. 1. CO2 reduction to CH4 was insensitive towards protonophores, when the Na+/H+ antiporter was inhibited; under these conditions delta mu Na+ was 120 mV (inside negative), whereas both delta mu H+ and the cellular ATP content were low. 2. CO2 reduction to CH4, rather than formaldehyde reduction, was sensitive towards Na+ ionophores, which dissipated delta mu Na+. 3. CO2 reduction to CH4, in the presence of protonophores and Na+/H+ antiport inhibitors, was coupled with the extrusion of 1-2 mol Na+/mol CH4, and formaldehyde reduction to CH4 was coupled with the extrusion of 3-4 mol Na+/mol CH4. Thus during CO2 reduction to the formaldehyde level 2-3 mol Na+ were consumed. PMID- 2557211 TI - Lactoferrin-binding sites at the surface of HT29-D4 cells. Comparison with transferrin. AB - The binding of 125I-lactoferrin to HT29-D4 cells, a clone of HT29 cells, was studied and compared to the binding of 125I-transferrin to the same cells. The binding of the two iron-transport proteins is saturable and reversible suggesting the presence of specific receptors for each protein. Scatchard analysis suggests the existence of binding sites for lactoferrin with the relatively high equilibrium dissociation constant, Kd1 of 408 nM. Additionally, the cell is capable of binding large amounts of lactoferrin with very low affinity, probably in a non-receptor intermediate fashion. The dissociation constant of transferrin and its receptor was calculated 9.29 nM which corresponds well to values found in the literature. In contrast to lactoferrin, the cell was capable of binding only low amounts of transferrin in a non-receptor intermediate fashion. After chemical crosslinking of lactoferrin to the cell surface, the radiolabeled lactoferrin was found in a complex of molecular mass 300 kDa. Crosslinking of transferrin resulted in a complex of much higher molecular mass. These data clearly show a binding site for lactoferrin different from the transferrin receptor. Only if competition experiments were performed with a high molar excess of both ligand proteins did a small percentage of either of the two ligands crossreact with the receptor for the other, possibly due to a structural similarity of the two glycoproteins. PMID- 2557212 TI - Identification of P2Y purinoceptors associated with voltage-activated cation channels in cardiac ventricular myocytes of the rat. AB - Extracellular ATP has vasodilatory and inotropic effects in the heart. We have demonstrated that extracellular ATP, in a concentration-dependent manner (10 nM 0.1 mM), increased [Ca2+]i in suspensions of isolated fura-2-loaded rat cardiac ventricular myocytes (maximum 96 +/- 10% increase over basal levels, SEM, n = 12, P less than 0.01). The increase in [Ca2+]i was often biphasic, with an initial fast phase (less than 1 s) of low amplitude, followed by a slower phase of higher amplitude. A second application of ATP had little effect, and ATP abolished the effect of subsequent electrical stimulations, even through the cells were still able to respond with an increase in [Ca2+]i to KCl-induced depolarization or stimulation by caffeine. Pretreatment of cells with nifedipine, verapamil, caffeine, ryanodine, or 8-(N,N-diethylamino)octyl 3,4,5-trimethoxybenzoate hydrochloride attenuated the effect of extracellular ATP on [Ca2+]i, and binding of extracellular free calcium by excess EGTA completely abolished the effects of extracellular ATP and electrical stimulation. Extracellular ATP increased bisoxonol fluorescence in ventricular myocytes, indicating depolarization of the sarcolemma. Pretreatment of the myocytes with tetrodotoxin (50 microM), or replacement of NaCl in the incubation buffer with the impermeant cation N-methyl D-glucamine, suppressed the extracellular ATP effect on [Ca2+]i. ADP and AMP had smaller effects on [Ca2+]i than ATP; adenosine had no effect. ATP analogues showed the following rank order of potency in increasing [Ca2+]i or bisoxonol fluorescence: ATP greater than or equal to 2-methylthioATP much greater than adenosine 5'-O-[3-thio]triphosphate greater than adenosine 5'-[alpha, beta methylene]triphosphate approximately adenosine 5'-[beta, gamma methylene]triphosphate approximately adenosine 5'-[beta, gamma-imino]triphosphate greater than adenosine. These data are consistent with the presence of purinoceptors (P2Y subtype) on the sarcolemma of cardiac ventricular myocytes of the rat, which upon activation lead to depolarization and activation of cation channels of the sarcolemma and flux of extracellular Ca2+ into the cells. This may result in further flux of Ca2+ into the cytosol from intracellular stores. The effects of extracellular ATP on [Ca2+]i in rat cardiac ventricular myocytes may, in part, explain the direct inotropic effects of extracellular ATP on the mammalian heart. PMID- 2557213 TI - A spin-label electron spin resonance study of the binding of mitochondrial creatine kinase to cardiolipin. AB - The binding of the mitochondrial creatine kinase to aqueous dispersions of beef heart cardiolipin has been studied via the perturbation of the mobility of spin labelled cardiolipin, using electron spin resonance (ESR) spectroscopy. In the presence of creatine kinase (1:1 protein/lipid ratio, by mass), the ESR spectra of cardiolipin labelled in a single acyl chain [n-(4,4-dimethyl-oxazolidinyl-N- oxy)stearoylcardiolipin] indicate a restriction of motion both at the C-5 and C 14 positions (n = 5, 14) of the lipid chains. The restriction in mobility was reversed by addition of phosphate or adriamycin, which are thought to inhibit the binding of creatine kinase to the mitochondrial membrane or to displace it from its binding site on the membrane. The effect of the protein on the chain mobility is consistent with surface binding of the protein; no positive evidence was obtained for penetration of the protein into the hydrophobic region of the membrane. PMID- 2557214 TI - Regulation of cytosolic aspartate aminotransferase mRNAs in the Fao rat hepatoma cell line by dexamethasone, insulin and cyclic AMP. AB - Glucocorticoid hormones increase the activity of cytosolic aspartate aminotransferase (cAspAT) in the Fao rat hepatoma cell line. Maximal increase (6 10-fold) was observed 48 h following the addition of the glucocorticoid agonist dexamethasone at a concentration of 0.1 microM. The effect of dexamethasone was specific since it was not mimicked by sex steroids and was inhibited by the glucocorticoid antagonist RU 486. Insulin (0.1 microM) inhibited by more than 50% the induction of cAspAT by glucocorticoids. The cAMP analog, 8-bromoadenosine 3',5'-monophosphate (Br8cAMP, 0.5 mM), potentiated the effect of dexamethasone (2 3-fold) and partially relieved the inhibitory effect of insulin on the induction by dexamethasone. Both insulin and Br8-cAMP had no significant effect on basal activity. The mitochondrial isoenzyme was insensitive to the various hormonal treatments. Northern blot analysis revealed the presence of two major (2.1-kb and 1.8-kb) and one minor (4-kb) mRNA species hybridizing with a rat cAspAT probe. The regulation of these mRNAs by glucocorticoids, insulin and cAMP correlated with the variation of the cAspAT activity, suggesting that these hormones act at the pretranslational level. We compared the regulation of cAspAT mRNAs with those of tyrosine aminotransferase mRNA. Both were similarly increased by dexamethasone but the latter was also increased by cAMP even in the absence of the glucocorticoid agonist. In addition, the increase in tyrosine aminotransferase mRNA was inhibited by cycloheximide whereas the increase in cAspAT mRNAs was not. These results show that there are significant differences in the regulation of cAspAT and tyrosine aminotransferase by glucocorticoids and other hormones, although both enzymes probably contribute to the same metabolic pathway. PMID- 2557215 TI - 131I-metaiodobenzylguanidine (mIBG) for bronchial oat cell cancer and melanoma detection? AB - The neuroendocrine features of bronchial oat cell carcinoma and melanoma indicate the possibility of positive imaging by means of radiolabelled metaiodobenzylguanidine. However, only four out of seven patients with bronchial oat cell carcinoma and three out of seven with melanoma were correctly diagnosed. Only false negative and no false positive results were obtained. The findings demonstrate a limited diagnostic value of the tracer in the tumor types examined. PMID- 2557216 TI - Comments on: "Two Routes for Renal 99mTc-DMSA Uptake into the Renal Cortical Tubular Cell". PMID- 2557217 TI - Plasma beta-endorphin, ACTH and cortisol secretion in man after nasal spray administration of calcitonin. AB - Beta-endorphin, ACTH and cortisol secretion were measured in twelve healthy adult males after nasal spray administration 200 IU salmon calcitonin. A significant increase in plasma beta-endorphin, from 19.2 ng/l under basal conditions to a peak of 27.1 ng/l at 30 min was recorded. Plasma ACTH and cortisol were not affected. In individual subjects the beta-endorphin level was increased in eight of the twelve, ACTH rose in three and cortisol did not change in any of them. The data indicate that calcitonin induced a beta-endorphin increase independent of enhanced corticotrophin-cortisol release. PMID- 2557219 TI - Adverse effects of three different forms of heparin therapy: thrombocytopenia, increased transaminases, and hyperkalaemia. AB - A prospective study has been made of the incidence of changes in transaminase levels, hyperkalaemia and thrombocytopenia in three groups of patients: 89 consecutive patients with venous thrombosis receiving therapeutic heparinization, 49 patients admitted because of hip fracture and receiving prophylactic low-dose conventional heparin, and 43 patients admitted because of hip fracture and randomly allocated to receive low molecular weight heparin. Laboratory measurements were made on admission and 8 days after commencing heparin. Only two patients on high-dose heparin developed thrombocytopenia. Increased transaminases were frequent with conventional heparin (18% and 32% of patients on high-dose heparin developed abnormal AsT and AlT values, respectively compared with 14% and 17% patients on low dose therapy). In contrast, only one patient on low molecular weight heparin developed abnormal AlT activity. Hyperkalaemia was uncommon in patients on any form of heparin therapy, and severe hyperkalaemia occurred in only one patient. PMID- 2557218 TI - New in vivo model to assess venous endothelial cell functions. Effect of defibrotide. AB - In the past few years there has been increasing interest in the role of the vascular endothelium as an active modulator of biological responses. Endothelial cells exert antithrombotic activity by the release of prostacyclin [23] and adenine nucleotides [16], the availability on the cell surface of heparin-like substances [3], and thrombomodulin-mediated activation of protein C [8]. In addition, endothelium is involved in the regulation of fibrinolysis by releasing soluble factors, such as tissue plasminogen activator (tPA; [10]) and plasminogen activator inhibitor (PAI; [22, 11]), as well as in the control of vascular responsiveness by the production of smooth muscle relaxing and contracting factors. Endothelial cells have also been shown to synthesize and to express procoagulant activities [18]. Many data on endothelial cell functions has been obtained from two experimental models, namely endothelial cell cultures and perfused segments of animal and human vessels. Both are subject to methodological criticism since they only represent in part in vivo conditions, and the necessary experimental manipulations and laboratory procedures greatly modify the naturally occurring cellular functions. In order to overcome such difficulties as far as possible, a new in vivo model has been employed to provide easily assessable and reliable data on the properties of endothelial cells in man. A venous segment was isolated functionally by cannulating a dorsal vein in the hand and a cubital vein in the same arm. Changes observed ex vivo in blood from the cubital vein following infusion into the hand vein of an active drug, can mainly be attributed to its local effect on the venous wall. At the same time, a cubital vein in the other arm was cannulated in order to provide information to distinguish systemic from regional effects. PMID- 2557220 TI - Decreased plasma membrane fluidity of peripheral blood lymphocytes after diethyldithiocarbamate (DTC) therapy in HIV-infected patients. AB - In seeking the putative mechanism of action of diethyldithiocarbamate (DTC) on the immune status of HIV infected patients, the plasma membrane fluidity of peripheral blood lymphocytes (PBL) from DTC-treated and untreated patients (CDC III-IVc1) was determined. Anisotropy values of the fluorescent probe 6-(9 anthroyloxy) stearic acid were increased in DTC-treated patients (0.175 vs 0.161), indicating decreased PBL plasma membrane fluidity. The membrane rigidifying effect was significantly greater 4 h after i.v. drug administration (0.185 in treated patients). As the membrane fluidity and the function of membrane embedded antigen are interdependent, it is possible that alterations in biophysical and/or biochemical properties of membranes may account for the beneficial effect of DTC on the immune function and clinical status of HIV infected patients. PMID- 2557221 TI - Analysis of signaling via surface immunoglobulin receptors on B cells from CBA/N mice. AB - CBA/N mice, which carry the xid immunodeficiency, lack a mature subpopulation of B cells. The residual B cells in these mice do not make antibodies to type-2 T independent antigens, nor do they synthesize DNA in response to mitogenic forms of anti-Ig antibodies. It is therefore an attractive hypothesis that the surface immunoglobulin receptors (sIgR) on xid B cells signal abnormally following cross linking. We show here that anti-Ig antibodies do cause inositol phospholipid hydrolysis and Ca2+ mobilization in xid B cells. However, the response of these cells are only 40%-50% of those of normal B cells. Studies with permeabilized cells demonstrated that the hyporesponsiveness is not due to ineffective coupling of sIgR to their associated G-protein. Rather it is apparently due to a quantitative and/or qualitative deficiency in the polyphosphoinositide-specific phosphodiesterase which mediates sIgR-induced inositol phospholipid hydrolysis. These observations may provide a biochemical explanation for the immunological abnormalities resulting from the xid mutation. PMID- 2557222 TI - Inhibition of cisplatin-induced emesis in the pigeon by a non-psychotropic synthetic cannabinoid. AB - The (+) enantiomer of the synthetic cannabinoid, 7-hydroxy-delta-6 tetrahydrocannabinol, dimethylheptyl homolog (HU-211), possesses significant antimetic efficacy in the pigeon. However, unlike all anti-emetic cannabinoids tested in the past, it is devoid of psychotropic (cannabimimetic) activity. The anti-emetic activity of HU-211 was determined in pigeons given 10 mg/kg i.v. cisplatin, a widely used antitumour agent, which is also a potent emetogenic agent at this dose. This activity was compared with that of delta-1 tetrahydrocannabinol (delta-1-THC). HU-211 pretreatment elicited a dose-related inhibition of cisplatin vomiting, with the optimal dose of HU-211 (2.5 mg/kg) inhibiting emesis by nearly 90%. Delta-1-THC in doses up to 5 mg/kg caused only an insignificant reduction in vomiting. The activity was increased in the presence of cupric chloride (0.8 mg/kg). The optimal dose of delta-1-THC (5.0 mg/kg) with CuCl2 very significantly diminished the total amount of vomitus expelled (up to 90%). However, it failed to inhibit emesis in 50% of all animals tested, did not significantly affect the time of onset of emesis and was highly psychotropic. The optimal dose of HU-211 (2.5 mg/kg) with CuCl2 inhibited emesis by 97%, significantly delayed the time on onset of emesis in the very few animals that did vomit and was completely non-psychotropic. The curve for the antiemetic effect of HU-211 was U-shaped over a narrow dose range. The present report demonstrates that complete separation of psychotropic and antiemetic activities is possible in the cannabinoid series. PMID- 2557223 TI - Modulation of mu-mediated antinociception by delta agonists: characterization with antagonists. AB - The functional interactions between supraspinal mu and delta receptors were characterized in the mouse using mu receptor-selective antagonists. The effects of pretreatment with the mu opioid antagonists, beta-funaltrexamine (beta-FNA) and naloxonazine on the modulation of morphine antinociception by the delta agonists [D-Pen2,D-Pen5]enkephalin (DPDPE) and [D-Ala2,Met5]enkephalinamide (DAMA) were studied. When co-administered in the same i.c.v. injection, a sub antinociceptive dose of DPDPE consistently and significantly increased the antinociceptive potency of morphine in control animals, while a sub-effective dose of DAMA decreased morphine antinociception; both the respective increase and the decrease of morphine potency by DPDPE and DAMA had been previously shown to be blocked by ICI 174,864, a delta antagonist. Pretreatment of mice with the non equilibrium mu antagonist beta-FNA 4 h prior to testing, a pretreatment which had no effect on i.c.v. DPDPE or DAMA antinociception, prevented the modulation of morphine antinociception by both DPDPE and DAMA. Pretreatment with the long acting mu 1 antagonist naloxonazine, 24 h prior to testing, failed to affect the modulation of morphine antinociception by either DPDPE or DAMA; such a pretreatment had no effect on the antinociceptive effects of DPDPE or DAMA when given alone. These results provide further support for the concept of a functionally coupled mu-delta receptor complex which is sensitive to antagonism by beta-FNA, but not naloxonazine, and support the notion that subtypes of opioid mu and delta (i.e. complexed and non-complexed) receptors may exist. PMID- 2557224 TI - Myocardial Na+-K+-ATPase activity and [3H]ouabain binding sites in hypertensive rats. AB - Na+-K+-ATPase activity and [3H]ouabain binding were studied in cardiac ventricles of single wrapped kidney and DOCA-NaCl hypertensive rats. It was found that the total Na+-K+-ATPase activity decreased in the DOCA-NaCl and kidney wrapped hypertensive rats. The decrease of enzyme activity in DOCA-NaCl hypertensive rats was due to extracellular fluid expansion induced by NaCl loading, as DOCA itself had no effect on the enzyme. All these alterations were specific for Na+-K+ ATPase, since Mg2+-ATPase and 5'-nucleotidase activities were unaffected. Binding studies with [3H]ouabain showed that the decrease in Na+-K+-ATPase activity was due to a reduction in the number of binding sites for ouabain rather than to a change of binding affinity. The reduced myocardial Na+-K+-ATPase activity observed in these two types of low renin hypertension, coupled with the observation of reduced vascular Na+ pump activity by others, suggests a common underlying defect in the cardiovascular Na+-K+ transport system of these hypertensive rats. PMID- 2557225 TI - Paradoxical response of plasma beta-endorphin to combined administration of TRH and GnRH in adrenal disorders. AB - The combined intravenous injection of TRH and GnRH elicited paradoxical responses of plasma beta-endorphin in active and successfully treated pituitary dependent Cushing's disease as well as in ectopic ACTH syndrome and in congenital adrenal hyperplasia. No response was observed in Cushing's syndrome due to adrenal tumours. It is concluded that an abnormal response to inappropriate releasing hormones cannot verify the existence of a pituitary corticotrophic microadenoma. PMID- 2557226 TI - Effect of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TTPA) upon membrane ionic exchanges in sea urchin eggs. AB - The effect of TPA (12-O-tetradecanoylphorbol-13-acetate) upon ionic exchanges was investigated in eggs of the sea urchin Arbacia lixula. Ouabain-sensitive 86Rb uptake and amiloride-sensitive 24Na influx were dramatically stimulated after TPA addition, indicating an enhancement of total ionic permeabilities. Stimulation by TPA of both Na+/H+ and Na+/K+ exchanges was canceled by amiloride, suggesting that activation of protein kinase C elicits, via Na+/H+ activity, stimulation of the sodium pump. However, TPA did not stimulate sodium pump activity and Na+/H+ exchange at the same rate as fertilization, probably because of an absence of calcium-dependent events. Further fertilization of TPA-pretreated eggs triggered an enhancement of sodium pump activity when the TPA treatment duration did not exceed 10 min. It is suggested that TPA activates preexisting transporting mechanisms in plasma membranes of unfertilized eggs (Na+ pump, Na+/H+ exchange) without eliciting corresponding regulatory mechanisms (Na+ stat, pH stat). PMID- 2557228 TI - Close relationship between modulation of serum-induced stimulation of DNA synthesis and changes in gap-junctional intercellular communication in quiescent 3T3-L1 cells caused by cyclic AMP and the tumor-promoting phorbol ester TPA. AB - Involvement of gap-junctional intercellular communication in the stimulation of growth was investigated in quiescent 3T3-L1 cells. When the cells in monolayer were growth-arrested by culture in a low concentration of calf serum, addition of dibutyryl cyclic AMP enhanced dye-coupling and suppressed the enhancement of DNA synthesis, induced by calf serum, in quiescent cells. 12-O-Tetradecanoylphorbol 13-acetate (TPA) suppressed dye-coupling in quiescent cells and enhanced DNA synthesis in both quiescent and serum-treated cells. When about 5000 cells were cultured in contact to form a colony, growth arrest of the cells was observed in the central region of such colonies rather than in the peripheral region, but addition of calf serum induced DNA synthesis in the cells in both the peripheral and central regions of the colonies. Addition of TPA enhanced serum-induced DNA synthesis in the cells in the central region of colonies rather than in the peripheral region. These results suggest that the ability of quiescent cells to escape from growth arrest is inversely correlated to the extent of gap-junctional intercellular communication. PMID- 2557227 TI - Adrenergic regulation of c-fos expression in cultured BC3H1 muscle cells. AB - Administration of adrenergic agonists induced c-fos mRNA in the salivary glands of the mouse and in the heart of the mouse, rat, and hamster (Barka et al., 1986, Mol. Cell Biol. 6, 2984-2989; 1987; Oncogene 1, 439-443). To further analyze transcriptional and post-transcriptional control of c-fos expression by adrenergic receptors and the putative role of fos in replication and differentiation pathways, we have examined c-fos expression in BC3H1 cells, a tumor-derived nonfusing muscle cell line. BC3H1 cells possess alpha 1- and beta 2 adrenergic receptors as well as receptors for histamine and acetylcholine. Furthermore, rapidly proliferating BC3H1 cells undergo differentiation toward muscle phenotype when exposed to low serum-containing culture media. Both alpha- and beta-adrenergic agonists and the tumor promoter 12-O-tetradecanoylphorbol-13 acetate caused a rapid, transient increase in the steady-state level of c-fos mRNA. This induction was essentially independent of whether the cells were in the proliferative, relatively quiescent, or differentiated state. Protein synthesis inhibitors cycloheximide and anisomycin also increased markedly the concentration of c-fos mRNA, and in the presence of anisomycin c-fos mRNA was superinduced by the alpha-adrenergic agonist norepinephrine. Run-on transcription assays indicated that the c-fos gene is expressed in both proliferating and differentiated cells, although the steady-state levels of c-fos mRNA were low, or even undetectable, in such cells. The adrenergic agonists and the tumor promoter stimulated the transcription of the c-fos gene in both proliferating and differentiated cells. This stimulation, however, was modest, two- to three-fold compared to controls, in contrast to the marked elevation of the level of c-fos mRNA they caused. Neither the proliferation nor the expression of muscle type creatine kinase activity was influenced by adrenergic agonists. It is suggested that activation of the c-fos gene is a consequence of adrenoreceptor stimulation in diverse cell types, and thus it is involved in pleiotropic cellular responses to adrenergic agonists. Catecholamines may be one of the physiologic regulators of the c-fos gene. PMID- 2557229 TI - Phorbol esters inhibit chondrogenesis in limb mesenchyme by mechanisms independent of PGE2 or cyclic AMP1. AB - Effects of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) on chondrogenesis and concentrations of prostaglandin E2 (PGE2) and cyclic AMP (cAMP) were investigated in micromass cultures of chick limb mesenchyme derived from the distal tip of stage 25 limb buds. TPA completely inhibited chondrogenesis during the first 4 days of culture; however, a few small cartilage nodules formed by day 6. Relative to control cultures, both PGE2 and cAMP concentrations were altered by TPA treatment during the 6-day period of cell culture. Concentrations of both compounds increased in control cells during the first 24 h of culture and then declined during the remaining 5 days. In TPA treated cells both PGE2 and cAMP levels increased progressively during the 6 days of days of cell culture, each being elevated at day 6 by twofold over control cells. The results suggest the presence of regulatory pathways important in chondrogenesis which occur independent of those initiated by PGE2 and the cAMP system. PMID- 2557231 TI - VIP receptors and control of short circuit current in the human intestinal clonal cell line Cl.19A. AB - At the maximally effective concentration of 10 nM, VIP induced a marked (12.5 fold stimulation above basal), and sustained increase in short circuit current in the human intestinal epithelial cell line Cl.19A grown on permeable filters and placed in Ussing chambers. Half-maximal increase of Isc was observed for 0.1 nM VIP. This was well correlated with the VIP-stimulated adenylate cyclase activity (ED50:0.07 nM). Binding studies using 125I-VIP indicated that Cl.19A cells express a peptide-specific VIP receptor with a dissociation constant of 0.07 nM. Covalent labeling of receptors followed by SDS-PAGE analysis of membrane proteins resulted in the identification of a 63,000 dalton binding protein in Cl.19A cells. PMID- 2557230 TI - Paraventricular nucleus neuronal responses following electrical stimulation of the midbrain dorsal raphe: evidence for cotransmission. AB - In order to determine the responses of paraventricular nucleus neurones following activation of central serotonergic pathways, single unit activity was recorded and responses following electrical stimulation of the midbrain dorsal raphe nucleus were examined. Excitation was recorded from approximately 50% of the cells, independent of whether they were antidromically identified as projecting to the median eminence or unidentified. Approximately 20% of cells were inhibited by the stimulation, the majority of these being unidentified. Parachlorophenylalanine-induced inhibition of serotonin synthesis reduced hypothalamic serotonin levels by 77% and caused a significant reduction in the proportion of cells excited by the stimulation, whereas the inhibitory responses were not affected. Intracerebroventricular administration of the serotonergic neurotoxin, 5,7-dihydroxytryptamine, which caused similar reductions in hypothalamic serotonin content (77%), reduced still further the proportion of excitatory responses and also reduced the proportion of cells inhibited by the stimulation. The data obtained suggest that serotonin acts as an excitatory neurotransmitter in the paraventricular nucleus; this is discussed particularly with respect to the regulation of the hypothalamo-hypophysial-adrenocortical axis. The loss of inhibitory responses in 5,7-dihydroxytryptamine treated, as opposed to the parachlorophenylalanine treated, animals suggests that the serotonergic fibers innervating the recorded cells may contain a cosecreted substance that may have important physiological actions in the control of neuronal activity in the region recorded. PMID- 2557232 TI - Tissue-specific expression of c-Ha-ras in premalignant gastrointestinal mucosae. AB - The molecular mechanisms underlying premalignant gastrointestinal diseases, such as ulcerative colitis and Barrett's esophagus, remain unknown. For this reason, the expression of the protooncogene c-Ha-ras was studied in ulcerative colitis and Barrett's esophagus. Total cellular RNA was extracted from different regions of the gastrointestinal tract in these two diseases. Expression of c-Ha-ras was greater in proximal than in distal colon and undetectable in Barrett's esophagus. These regional differences in expression were not seen with the control gene beta actin or with the protooncogenes c-myc and p53. In order to evaluate structural factors contributing to expression, amplification and methylation of c-Ha-ras DNA were studied in these tissues by Southern and slot blotting. No amplification of c-Ha-ras or six other protooncogenes was detected. These data suggest tissue specific regulation of c-Ha-ras expression in the gastrointestinal tract in certain premalignant disease states. PMID- 2557234 TI - [Characteristics of the action of euphylline and the activity of forms of cAMP phosphodiesterase in lymphocytes of patients with bronchial asthma]. AB - Two forms of cAMP phosphodiesterase were isolated from lymphocytes by using centrifugation in sucrose density gradient. The activities of both forms do not differ in healthy persons and in patients with bronchial asthma during asphyxia free period. During the attack the activity of the high molecular form of the enzyme increases. Euphylline exerts the inhibitory effect on the activity and consequently on the general activity of the enzyme. PMID- 2557233 TI - [The action of fenigidin (nifedipine) on central hemodynamics and cerebral blood flow in hypertension]. AB - Nifedipine is an effective agent in hypertensive emergencies as well as in the long-term management of hypertension especially for the patients with an increased risk of cerebral hypoperfusion. Nifedipine exerts its blood lowering effect by reducing total peripheral resistance including cerebrovascular resistance. Despite the marked reduction of perfusion pressure, the cerebral blood flow is maintained by nifedipine. PMID- 2557235 TI - Amplification of the phosphorylation site-ATP-binding site cDNA fragment of the Na+,K(+)-ATPase and the Ca2(+)-ATPase of Drosophila melanogaster by polymerase chain reaction. AB - In vitro DNA-amplification technique has been utilized to generate a 430 bp fragment of the Na+,K(+)-ATPase, and a 550 bp fragment of a Ca2(+)-ATPase (the sarcoplasmic reticulum-type) of Drosophila melanogaster. The oligonucleotide primers for the DNA-amplification (Polymerase Chain Reaction) had been designed on the basis of amino acid sequence motifs--the phosphorylation site and the ATP binding site--conserved among members of the ATPase protein family. Using the amplified cDNA-segments as probes, we demonstrated that there is one Na+,K(+) ATPase and one Ca2(+)-ATPase (sarcoplasmic reticulum-type) gene in the Drosophila genome. Three different mRNA species are processed from the Na+,K(+)-ATPase gene and one from the Ca2(+)-ATPase gene. Developmental control in expression of the Ca2(+)-ATPase gene was observed. PMID- 2557236 TI - Stimulation of FA and phosphatase-1 activities by insulin in 3T3-L1 cells. AB - The phosphatase-1 activator FA and phosphatase-1 were assayed in 3T3-L1 cells exposed to insulin. The cytosolic FA activity was transiently stimulated (7-8 fold) 1 and 2 min after exposure to 10(-8) M insulin and returned to control values within 5-10 min. Cytosolic phosphatase-1 (assayed after trypsin treatment) was activated (120-140% of controls) between 2 and 5 min and returned to control values within 10 min. Insulin effects were dose-dependent, with maximum stimulation of both activities at 10(-8) M insulin. The possibility that FA and other kinases mediate phosphatase activation by insulin is discussed. PMID- 2557237 TI - HSV infection induces increased transcription of Alu repeated sequences by RNA polymerase III. AB - The Alu family of repeated sequences is transcribed by both RNA polymerase II and RNA polymerase III. In cells infected with HSV, transcription by polymerase III increases while transcription by polymerase II decreases. By using virus strains carrying mutations in the genes encoding individual regulatory proteins, we have shown that this effect is dependent upon the immediate-early protein ICP27 and occurs by a process distinct from those which regulate viral gene expression. This is the first example of increased transcription of endogenous cellular sequences by RNA polymerase III during infection with a DNA virus. PMID- 2557238 TI - Modification of the electron-transfer sites of Pseudomonas aeruginosa azurin by site-directed mutagenesis. AB - Site-directed mutagenesis of the structural gene for azurin from Pseudomonas aeruginosa has been used to prepare azurins in which amino acid residues in two separate electron-transfer sites have been changed: His-35-Lys and Glu-91-Gln at one site and Phe-114-Ala at the other. The charge-transfer band and the EPR spectrum are the same as in the wild-type protein in the first two mutants, whereas in the Phe-114-Ala azurin, the optical band is shifted downwards by 7 nm and the copper hyperfine splitting is decreased by 4.10(-4)/cm. This protein also shows an increase of 20-40 mV in the reduction potential compared to the other azurins. The potentials of all four azurins decrease with increasing pH in phosphate but not in zwitterionic buffers with high ionic strength. The rate constant for electron exchange with cytochrome c551 is unchanged compared to the wild-type protein in the Phe-114-Ala azurin, but is increased in the other two mutant proteins. The results suggest that Glu-91 is not important for the interaction with cytochrome c551 and that His-35 plays no critical role in the electron transfer to the copper site. PMID- 2557239 TI - Role of supernumerary subunits in mitochondrial cytochrome c oxidase. AB - Role of supernumerary subunits of bovine heart cytochrome c oxidase has been investigated by examining the influence on the enzymatic activity of their removal by chromatographic procedures or controlled digestion by trypsin. Is has been shown that partial proteolytic cleavage of subunit IV results in depression of respiratory activity and of redox-linked proton translocation. Selective removal by gel-filtration of subunit Vlb has no significant influence on the redox and protonmotive activity of the oxidase. PMID- 2557240 TI - Na+/H+ exchange and Ca2+ influx. AB - Cell stimulation raised the cytosolic free Ca2+ concentration, [Ca(2+]i and induces activation of Na+/H+ exchange which raises the cytosolic pH, pHi. Recent studies have addressed the question whether Na+/H+ exchange plays a role in Ca(2+) influx and, specifically, whether a rise in pHi alone suffices to open Ca(2+) channels in the plasma membrane. Artificial cytosolic alkalinization can induce Ca(2+) uptake across the plasma membrane of endothelial cells, lymphocytes and smooth muscle cells. Furthermore, inhibition of Na+/H+ exchange reduces agonist-induced Ca(2+) influx in endothelial cells and platelets which supports the concept that pHi may regulate the opening of Ca(2+) channels in the plasma membrane. Although these findings argue in favour of a role of Na+/H+ exchange in Ca(2+) influx, the onset of pHi and Ca(2+) rises, measured with fluorescent indicators, suggests that the increase in [Ca(2+)]i distinctly precedes the increase in pHi. This challenges the concept that alkalinization per se is a sufficient signal for the opening of Ca(2+) channels in the plasma membrane. PMID- 2557241 TI - Nuclear envelope assembly around sperm chromatin in cell-free preparations from Drosophila embryos. AB - Chicken sperm chromatin initiated an assembly of interphase-like nuclei in a cell free cytoplasmic preparation from 1-6 h old Drosophila melanogaster embryos. The formation of these interphase-like nuclei from the condensed sperm chromatin happened in a series of distinct steps. Anti-Drosophila lamin monoclonal antibody stained the assembled nuclei in a pattern indistinguishable from normal Drosophila nuclei. This assembly process required an ATP regenerating system and could be blocked by the addition of novobiocin into the cell-free extract. PMID- 2557242 TI - Loss of calcium sensitivity of plasma gelsolin is associated with the presence of calcium ions during preparation. AB - Gelsolin is a calcium-dependent actin severing and capping protein. Calcium 'opens' the molecule to make actin binding sites accessible, but removal of calcium from the medium does not necessarily fully reverse this process. The calcium sensitivity of actin monomer binding and actin filament severing is here shown to vary considerably with the source of gelsolin and conditions of preparation. Plasma gelsolin undergoes irreversible loss of calcium sensitivity when prepared in the presence of calcium ions. This is not due solely to effects of bound calcium, because purified human plasma gelsolin expressed in E. coli and stored in calcium shows no comparable loss of calcium sensitivity when prepared or stored in calcium. These results suggest the presence of factors in plasma which, in the presence of calcium, promote an irreversible structural change in gelsolin resulting in permanent loss of calcium sensitivity. PMID- 2557243 TI - A single point mutation confers tetrodotoxin and saxitoxin insensitivity on the sodium channel II. AB - A single point mutation of the rat sodium channel II reduces its sensitivity to tetrodotoxin and saxitoxin by more than three orders of magnitude. The mutation replaces glutamic acid 387 with a glutamine and has only slight effects on the macroscopic current properties, as measured under voltage-clamp in Xenopus oocytes injected with the corresponding cDNA-derived mRNA. PMID- 2557245 TI - Protein kinase C phosphorylates DNA topoisomerase I. AB - The induction of mammalian cell proliferation requires the expression of a specific set of genes. Tumor promoters stimulate cell growth by activating the Ca2+ and phospholipid-dependent protein kinase, protein kinase C (PKC). DNA topoisomerase I, a nuclear enzyme involved in transcription, was phosphorylated by activated PKC in vitro. Phosphorylation by PKC stimulated the DNA relaxation activity of topoisomerase I two- to three-fold. Therefore, DNA topoisomerase I is a substrate for PKC-mediated activation by phosphorylation and may serve as a nuclear target of mitogenic signals generated by tumor promoters in vivo. PMID- 2557244 TI - Functional expression of the calcium release channel from skeletal muscle ryanodine receptor cDNA. AB - Combined patch-clamp and fura-2 measurements were performed to study the calcium release properties of Chinese hamster ovary (CHO) cells transfected with the rabbit skeletal muscle ryanodine receptor cDNA carried by an expression vector. Both caffeine (1-50 mM) and ryanodine (100 microM) induced release of calcium from intracellular stores of transformed CHO cells but not from control (non transfected) CHO cells. The calcium responses to caffeine and ryanodine closely resembled those commonly observed in skeletal muscle. Repetitive applications of caffeine produced characteristic all-or-none rises in intracellular calcium. Inositol 1,4,5-trisphosphate (IP3) neither activated the ryanodine receptor channel nor interfered with the caffeine-elicited calcium release. These results indicate that functional calcium release channels are formed by expression of the ryanodine receptor cDNA. PMID- 2557246 TI - Histamine and bradykinin stimulate the phosphoinositide turnover in human umbilical vein endothelial cells via different G-proteins. AB - The G-proteins which regulate hormonal turnover of phosphoinositide (PI) in human umbilical vein endothelial cells have been investigated. A 40-41 kDa doublet present in the membranes of these cells was selectively ADP ribosylated by pertussis toxin (PTx), and this doublet was Gi alpha 2 and Gi alpha 3 according to immunoblotting with specific antisera. By contrast, a doublet of 24-26 kDa proteins in the same membrane preparations was ADP ribosylated by the C3 component of botulinum toxin (BoTx). PTx-dependent ADP ribosylation blocked stimulation of PI turnover by histamine, but did not affect stimulation by bradykinin, whereas BoTx (C2 + C3 components) had the opposite effect. Thus two different groups of G-proteins may be involved in hormone-dependent stimulation of PI turnover in human umbilical vein endothelial cells. PMID- 2557247 TI - Locoregional recurrences after 649 modified radical mastectomies: incidence and significance. AB - A continuous series of 649 patients, treated by modified radical mastectomy for primary breast carcinoma, is analyzed after a median follow-up of 92 months. 'True isolated' locoregional recurrences (LR), defined as LR not preceded or followed by distant metastases within 6 weeks, appeared with a cumulative actuarial incidence rate of 6%, 14% and 19% after 1, 5 and 7 years respectively, whereas the respective figures for distant metastases (M1) were 10%, 37% and 48%. The main initial parameters, predicting both the LR-free and the M1-free interval, are presented by statistical analyses in the following order of importance: number of invaded lymph nodes in the axilla, tumor size (T) and histological grading of differentiation. The same factors also predicted the imminence of M1 once LR had occurred, as well as survival after LR. A higher incidence of M1 after LR was also correlated with estrogen-receptor negative tumors and with those LR occurring within one year after mastectomy. LR occurred at the chest wall (65%), in the sub-clavicular fossa (16%) and the axilla (6%); the remaining 13% occurred in two of the sites. There was a trend towards longer survival after chest wall recurrence than after LR recurrence at another site. Axillothoracic irradiations postmastectomy gave a lower rate of LR in 227 patients than did a regimen of 12 months adjuvant chemotherapy with irradiation restricted to the internal mammary lymph nodes in 120 subsequent patients: 17 vs 25% at 5 years (P = 0.03 when adjusted by initial nodal involvement and T-size). Total excision of LR (repeated if new LR occurred) gave better rates of local ultimate control and survival than other kinds of treatments, with or without adjuvant local or systemic therapy. LR is not always a sign of imminent generalized disease. Actuarial 5-year survival after LR is 26.2% overall whereas, if only 'true isolated' LR are considered, the survival is 37%. PMID- 2557249 TI - New generation antibiotics--quinolones. PMID- 2557248 TI - [The multireceptor excitation of gastric secretion]. PMID- 2557250 TI - Account of an acute herpetic infection. PMID- 2557251 TI - Immunoglobulin and T cell receptor gene rearrangements in lymphoproliferative disorders. AB - Thirty-one samples representing Hodgkin's and non-Hodgkin's lymphomas, angioimmunoblastic lymphadenopathy (AILD), and benign follicular hyperplasia in HIV infections were examined for rearrangements of the immunoglobulin (Ig) and T cell receptor (TcR) beta-chain gene loci. In 11 of 12 non-Hodgkin's lymphomas (classified as Burkitt lymphoma (2), centrocytic lymphoma (1), centrocytic centroblastic lymphoma (5), centroblastic lymphoma (3], only rearranged Ig genes could be detected. The exceptional case was an unclassified high-grade lymphoma, which represented a rearrangement of the TcR beta-chain. We also examined DNA from lymphoid neoplasms in which the lineage of the malignant cell was still controversial. Rearrangement of the TcR could exclusively be demonstrated in all 3 cases of AILD. One Ig gene rearrangement and 4 TcR beta-chain rearrangements were found in 13 samples of Hodgkin's lymphomas (11 lymph nodes, 1 pleura effusion and 1 bone biopsy with proven infiltration). Examination of 3 cases of benign follicular hyperplasia in HIV infection represented one Ig rearrangement. PMID- 2557252 TI - Intellectual ability of children after perinatal cytomegalovirus infection. AB - The intellectual ability of 16 children who had not had congenital cytomegalovirus (CMV) infection, but acquired it before the age of six months, was compared with that of two other groups: one comprising 32 children who had had CMV-excreting mothers but who were not infected themselves: the other comprising 18 healthy controls. Serial audiological, ophthalmological and psychometric examinations were performed. The only statistically significant difference found was that the children with acquired CMV infection had lower mean arithmetic scores. The results suggest that healthy term infants who acquire early CMV infection are not at increased risk of intellectual impairment. PMID- 2557253 TI - [Immuno-neuroendocrine interactions during growth and aging]. PMID- 2557254 TI - Laparoscopy compared with ultrasonography in the diagnosis of hepatocellular carcinoma. AB - In recent years, various newer imaging procedures have superseded laparoscopy in the detection of many diseases. The role of laparoscopy in the diagnosis of hepatocellular carcinoma is still subject to debate. To assess the value of laparoscopy compared with that of ultrasonography, we compared data obtained from a series of 54 patients with hepatocellular carcinoma, all of whom had both procedures performed at nearly the same time. In our study, ultrasonography proved superior to laparoscopy in detecting the presence and extent of the hepatocellular carcinoma. Supplemental findings attributable to laparoscopy did not alter management, whereas additional information obtained by ultrasonography often did influence the choice of treatment. We conclude that ultrasonography is the primary diagnostic method of choice in the assessment of hepatocellular carcinoma and that laparoscopy should be reserved for only selected cases. PMID- 2557255 TI - Intraoperative fiberoptic choledochoscopy for malignant biliary tract obstruction. AB - A prospective study to determine the safety and effectiveness of intraoperative fiberoptic choledochoscopy in the management of malignant obstruction of the biliary tree was conducted in 44 patients. There were 12 patients with cancer of the pancreatic head, 9 with adenocarcinoma of the distal common bile duct, 4 with adenocarcinoma of the ampulla of Vater, 8 with cholangiocarcinoma of the common hepatic duct, 9 with intrahepatic bile duct carcinoma, and 2 with hepatocellular carcinoma. Five patients with intrahepatic bile duct carcinoma and two with hepatocellular carcinoma were found during a search for intrahepatic duct stones in patients with recurrent cholangitis. In nine patients with bile duct carcinoma (seven extrahepatic and two intrahepatic), histopathological diagnosis was made at operative endoscopy. Treatment strategy was altered in seven patients based on findings at operative choledochoscopy. Complications were few with no mortality. Intraoperative fiberoptic choledochoscopy was safe and useful in the management of malignant obstruction of the biliary tree. PMID- 2557257 TI - Laparoscopy and ultrasonography in the diagnosis of hepatocellular carcinoma. PMID- 2557256 TI - Delayed, massive hemorrhage following electrocoagulating biopsy ("hot biopsy") of a diminutive colonic polyp. PMID- 2557259 TI - Characterization of gonadotropin-releasing hormone binding sites in the pituitary of the three-spined stickleback, Gasterosteus aculeatus. AB - Binding sites for gonadotropin-releasing hormone (GnRH) in stickleback pituitary homogenates were characterized using an iodinated, superactive analog of salmon GnRH (sGnRH), D-Arg6-Pro9-sGnRH-NEt (sGnRHa). Binding of 125I-sGnRHa reached equilibrium after 60 min incubation at 4 degrees and was a function of tissue concentration. The specificity of 125I-sGnRHa binding was demonstrated by displacement with sGnRHa, sGnRH, and Buserelin [D-Ser(t-Bu)6-Pro9-GnRH-NEt]. Both Scatchard analyses of saturation data and displacement curves revealed a single class of high-affinity binding sites (Ka = 0.71 +/- 0.03 X 10(9) M-1, Bmax = 1087 +/- 165 fmol/mg protein). PMID- 2557258 TI - Fatal cerebral herniation from brain metastasis following fiberoptic endoscopy. PMID- 2557260 TI - The structural gene for aspartokinase II in Bacillus subtilis is closely linked to the sdh operon. AB - The aecA and aecB loci map at 250 and 290 degrees, respectively, on the Bacillus subtilis chromosomal genetic map. The aecB locus has been proposed as the structural gene for aspartokinase II. From DNA sequence analyses and comparisons to the sequence of the aspartokinase II gene, it can be concluded that the structural gene for aspartokinase II is located close to sdh at 250 degrees and cannot be aecB. A detailed map over 7 kbp in the 250 degree region is presented. PMID- 2557262 TI - Analysis of the Om(1D) locus in Drosophila ananassae. AB - From the ca;px stock, which is the progenitor of Om mutants caused by insertions of the tom retrotransposon, 50 kb of genomic DNA including the Om(1D) locus was cloned by tom tagging and chromosome walking. Southern blot analyses of six Om(1D) mutants exposed one or two tom elements inserted at five nonrandom sites within an 18-kb distal segment of the restriction map; the phenotypic uniformity between these mutants was not affected by variations in the position, number or orientation of their inserts. Spontaneous revertants or more extreme derivatives of Om(1D) alleles were nonlinearly associated with losses or gains of tom inserts. Seven of eight radiation induced derivatives of Om(1D) mutants had one breakpoint of a chromosome rearrangement in polytene section 13A which includes the Om(1D) locus. Two Om(1D) derivatives, a spontaneous revertant and an induced extreme allele, were associated with overlapping deficiencies which define a region that is likely to contain the Om(1D) coding seguences proximal to the tom insertion sites. Incidental results confirm the previously indicated homology of the Om(1D) locus with the Bar locus of Drosophila melanogaster. PMID- 2557263 TI - Genetic resistance to viral infection: the molecular cloning of a Drosophila gene that restricts infection by the rhabdovirus sigma. AB - The ref(2)P gene of Drosophila melanogaster has two common alleles, ref(2)Po which permits the infection of flies by the rhabdovirus sigma (sigma), and ref(2)Pp which is restrictive for sigma infection. This gene has been cloned by P element tagging and shown to code for two RNAs in adult flies. These RNAs are expressed in both males and females, but only the larger is expressed in ovaries. Both transcripts are shorter, by about 50 nucleotides, in flies carrying the ref(2)Pp allele than in those carrying ref(2)Po. The dominance relationships of these two alleles, and the fact that ref(2)Pnull alleles are permissive to sigma infection, suggest that the ref(2)Po product is antimorphic to that of the ref(2)Pp allele. PMID- 2557261 TI - Enhancement of Escherichia coli plasmid and chromosomal recombination by the Ref function of bacteriophage P1. AB - The Ref activity of phage P1 enhances recombination between two defective lacZ genes in the Escherichia coli chromosome (lac- x lac- recombination). Plasmid recombination, both lac- x lac- and tet- x tet-, was measured by transformation of recA strains, and was also assayed by measurement of beta-galactosidase. The intracellular presence of recombinant plasmids was verified directly by Southern blotting. Ref stimulated recombination of plasmids in rec+ and rec(BCD) cells by 3-6-fold, and also the low level plasmid recombination in recF cells. RecA independent plasmid recombination, either very low level (recA cells) or high level (recB recC sbcA recA cells), was not stimulated. Ref stimulated both intramolecular and intermolecular plasmid recombination. Both normal and Ref stimulated lac- x lac- chromosomal recombination, expected to be mostly RecBC dependent in wild-type bacteria, were affected very little by a recF mutation. We have previously reported Ref stimulation of lac- x lac- recombination in recBC sbcB bacteria, a process known to be RecF-dependent. Chromosomal recombination processes thought to involve activated recombination substrates, e.g., Hfr conjugation, P1 transduction, were not elevated by Ref activity. We hypothesize that Ref acts by unknown mechanisms to activate plasmid and chromosomal DNA for RecA-mediated recombination, and that the structures formed are substrates for both RecF-dependent (plasmid, chromosomal) and Rec(BCD)-dependent (chromosomal) recombination pathways. PMID- 2557264 TI - [Various types of structural organization of L1 transcripts in the rat brain]. AB - Recombinant cDNA clones homologous to L1 retroposon were isolated from the cDNA banks of rat brain. Their organisation was studied by sequencing and some resulting aspects are discussed. PMID- 2557265 TI - [Serological studies of cytomegalovirus in the Melut district, South Sudan]. AB - Out of 1071 investigated patients had antibodies in 73.7% against Cytomegalovirus. 99 ambulant Kala-Azar-patients reacted positive in 33%. Inhabitants of different villages have shown 35.2 to 50% positive reactions. Patients with hepato-splenomegaly in 94% and patients with diarrhoea and fever in 81% were positive. PMID- 2557266 TI - [Problems of hygienic standardization of electromagnetic fields produced by teletransmitting objects]. AB - Maximum allowable electromagnetic field levels produced by teletransmitting stations and differentiated by frequency have been described. The prospects of further studies on the improvement of hygienic standardization of electromagnetic fields have been set forth. PMID- 2557267 TI - [Study of the effect of microwave radiation on the nervous system (results of the Soviet-American Research Project)]. PMID- 2557268 TI - Perturbation of upper gastrointestinal transit and antroduodenal motility by experimentally applied stress: the role of beta-adrenoreceptor mediated pathways. AB - A series of three experiments were performed on healthy adult volunteers to investigate the possible role played by beta-adrenoreceptor mediated pathways in the disturbance of human upper intestinal motor function by hand immersion in cold water. In the first experiment, (an extended pilot study on one individual), orocaecal transit of a standard meal was measured on 36 occasions with and without cold water stimulation and with and without a series of alpha and beta blocking drugs. Cold water stimulation consistently delayed transit in this individual, an effect which was attenuated by prior beta-blockade. In a double blind trial of the effect of beta-blocker atenolol v placebo on transit in nine individuals, a consistent reduction in the cold water induced transit delay was observed (p less than 0.01) independent of any direct effect of beta-blockade. In the third experiment seven individuals underwent repeated studies of antroduodenal pressure activity comparing the effects of cold and warm water stimulation with and without beta blockade to determine whether the observed transit effect could be related to an action on gastrointestinal motility. Cold water stimulation reduced antroduodenal motility, but no consistent effects of previous beta blockade were noted. These studies indicate the presence of a beta adrenoreceptor mediated pathway in the cold water induced delay of orocaecal transit but not in the inhibition of gastroduodenal motility. Further studies are indicated to determine the site and mode of action of this transit effect more precisely. PMID- 2557269 TI - Is a persistent adenovirus infection involved in coeliac disease? AB - Recent evidence has implicated adenovirus 12 in the aetiology of coeliac disease so that persistent infection by this virus must be considered. We have undertaken a search for adenovirus DNA in duodenal biopsy samples from a total of 26 coeliac and non-coeliac patients. We could find no evidence of persistent virus DNA by Southern blot techniques even under conditions which approach a sensitivity of one copy of virus genome per cell, and use either adenovirus 12 or 41 DNA. PMID- 2557270 TI - Human papillomavirus associated with adenocarcinoma and adenosquamous carcinoma of the cervix: analysis by in situ hybridization. AB - The incidence of adenocarcinoma of the cervix appears to be increasing. Recent reports have demonstrated an association between adenocarcinoma of the cervix and human papillomavirus (HPV) by Southern blot hybridizations. In situ deoxyribonucleic acid (DNA) hybridization was performed on paraffin-embedded specimens to localize the source of HPV DNA. In pure adenocarcinoma five of six specimens were positive for HPV DNA. Four specimens contained HPV type 18 and one HPV type 16. Only one of three adenosquamous lesions was positive and it contained both HPV types 16 and 31. These findings suggest an association between HPV and adenocarcinoma of the cervix. PMID- 2557271 TI - Hypercalcemia in association with mucinous adenocarcinoma of the ovary: a case report. AB - Hypercalcemia in association with mucinous adenocarcinoma of the ovary has not previously been reported. Such a case is presented. PMID- 2557273 TI - [Etiology of premenstrual syndrome]. PMID- 2557272 TI - Vulvar adenosquamous carcinoma arising in a hidradenoma papilliferum, with rapidly fatal outcome: case report. AB - A 65-year-old woman with adenosquamous carcinoma of the vulva arising in a hidradenoma papilliferum died from disseminated tumor 2 months after local excision. The light and electron microscopic features of the lesion are presented. The possible origins of vulvar adenosquamous carcinoma are discussed. PMID- 2557274 TI - [Nedocromil sodium therapy in asthma patients. Therapeutic effect in addition to treatment with oral theophylline and inhaled bronchodilator agents]. AB - 131 asthmatics aged 12-65 years, who still had symptoms despite inhaled or oral bronchodilators, were included in this double-blind group comparative study involving nedocromil sodium (2 puffs of 2 mg each twice daily) and placebo (2 puffs twice daily). The study was carried out at 8 centers over six weeks. Under nedocromil sodium, cough, dyspnea and severity of attacks were reduced significantly. Overall patient assessment also clearly favoured the active substance. Nedocromil sodium was also superior to placebo in terms of the improvement of lung function (FEV1, FVC and PEFR). 26 patients complained of unusual symptoms (12 under nedocromil sodium, 14 under placebo). Nedocromil sodium proved to be an effective, safe and well-tolerated drug in the antiinflammatory long-term treatment of reversible obstructive airways disease. PMID- 2557275 TI - Immunohistochemical study of monoclonal antibody MGD-1 in gastric carcinoma. AB - This paper reports a pathological and immunohistochemical study of gastric carcinoma for immunoreactivity with a monoclonal antibody. MGD-1, raised against cells from an adenocarcinoma of stomach. Fifty-four of 61 gastric carcinomas (89%) were positive for MGD-1. Metastatic gastric carcinoma in local nodes was positive in all 11 such cases. Out of 40 examples of chronic atrophic gastritis, only three, with mild dysplasia, were positive (7.5%). Forty cases with normal gastric mucosa were negative. The MGD-1 detection-rate of well- and poorly differentiated gastric carcinoma was 85% and 93% respectively. The metastatic cells and cells infiltrating the submucosa and muscular layer were more frequently positive and showed stronger staining with MGD-1 than those in mucosa. These results show that MGD-1 possesses a high degree of specificity for gastric carcinoma and could be used diagnostically. PMID- 2557276 TI - Primary hepatocellular carcinoma in hereditary haemorrhagic telangiectasia: a case report and literature review. PMID- 2557278 TI - Prostaglandins modulate central serotonergic neurotransmission. PMID- 2557277 TI - The histogenesis of mammary and extramammary Paget's disease. PMID- 2557279 TI - Binding of phytohemagglutinin to bovine B lymphocytes and its role in stimulation of expression of bovine leukemia virus genome. AB - Phytohemagglutinin (PHA) is known to increase the synthesis of bovine leukemia virus (BLV) particles and viral antigens in short-term culture of BLV-infected neoplastic and non-neoplastic lymphocytes. This stimulation of BLV expression has been shown to be due to enhanced transcription of the viral genome by a PHA induced protein. We have investigated the binding of 125I-labelled PHA to BLV infected bovine B lymphocytes and subsequent events that may lead to the stimulation of BLV-p25 synthesis. We found that PHA binding to the infected cells were rapid, but only a small fraction of the bound PHA is translocated to nucleus. However, bound PHA dissociated rapidly from the cell membrane, but not from the nucleus when PHA is removed from the culture medium. Furthermore, continuous presence of PHA was not essential for optimal stimulatory activity, instead a minimum incubation with PHA for 6 hr. followed by culturing the infected cells in its absence, was sufficient to exhibit maximal stimulatory activity. Our results raise the possibility that interaction of PHA with the plasma membrane probably triggers synthesis of a protein which in turn enhances the transcription of BLV genome in vitro. PMID- 2557281 TI - Effects of cyclophosphamide on pigs infected and vaccinated with swinepox virus. AB - Piglets treated with cyclophosphamide (50 mg/kg body wt, iv) before infection/vaccination with swinepox virus showed a strong suppression of humoral immune response. Cell mediated immune response was also affected to some extent and the cyclophosphamide treated piglets did not resist the challenge on 21 day post infection/vaccination and developed mild lesions. PMID- 2557282 TI - Histologic evaluation of a retrieved human HA-coated subperiosteal implant: report of a case. AB - A retrieval study of a hydroxyapatite-coated subperiosteal implant, after 1 year of function, demonstrates a direct connection with the underlying bone. PMID- 2557280 TI - Effect of dietary fiber from banana (Musa paradisiaca) on metabolism of carbohydrates in rats fed cholesterol free diet. AB - Effect of feeding isolated dietary fiber from M. paradisiaca on the metabolism of carbohydrates in the liver has been studied. Fiber fed rats showed significantly lower levels of fasting blood glucose and higher concentration of liver glycogen. Activity of glycogen phosphorylase, glucose-1-phosphate, uridyl transferase and glycogen synthase was significantly higher while phosphoglucomutase activity showed lower activity. Activity of some glycolytic enzymes, viz. hexokinase and pyruvic kinase was lower. Glucose-6-phosphatase showed higher activity while fructose 1-6 diphosphatase activity was not affected. Glucose-6-phosphate dehydrogenase on the other hand showed higher activity. The changes in these enzyme activities have been attributed due to the effect of higher concentration of bile acids produced in the liver as a result of feeding fiber. Evidence for this has been obtained by studying the in vitro effect of cholic acid and chenodeoxy cholic acid. PMID- 2557283 TI - Quantification of bone in dental implant sites after composite grafting of the mandible: report of a case. AB - The amount of viable bone in a composite graft that consisted of autogenous iliac bone marrow, allogenic rib, and porous hydroxyapatite was quantified using standard histologic techniques and an optical insert disk prior to placement of endosteal implants in an augmented mandible. It is suggested that this technique could be used in such cases before deciding whether to proceed with implant insertion. PMID- 2557284 TI - Hypophosphatemic vitamin D resistant rickets treatment with one alpha hydroxy vitamin D3. PMID- 2557285 TI - Nesidioblastosis in the newborn. PMID- 2557286 TI - Japanese encephalitis in children in Bellary Karnataka. AB - One hundred and fifty cases of Japanese encephalitis (JE) in children below 12 years of age admitted to the Headquarters Hospital, Bellary Medical College during October, 1986 to January, 1987 were studied. The youngest child affected was 1 year old. The ratio of male to female was 2:1. Hindu patients (93.33%) were significantly more than Muslim patients (6.6%). The average duration of illness prior to hospitalisation was 4.2 days (SD +/- 1.9 days). Fever, headache and/or vomiting were common presenting symptoms at the onset of illness. Onset of illness was acute or sub-acute in 105 cases (70%). CSF examination showed pleocytosis with lymphocytic reaction and normal sugar. Japanese encephalitis virus was isolated from the brain biopsy tissue in one case and the seropositivity rate for JE was 50%. The mortality was 26% and was directly related to younger age, longer duration of illness prior to admission and deeper grade of coma at the time of admission. Moderate to severe sequalae were seen in 31 patients (28.82%). The mean duration of hospital stay was 8 days. PMID- 2557287 TI - From theory to practice: the planned treatment of drug users. Interview by Stanley Einstein. PMID- 2557288 TI - Suppressive activity of alveolar macrophages and blood monocytes from interstitial lung diseases: role of released soluble factors. AB - Two groups of patients suffering from interstitial lung diseases (ILD) namely sarcoidosis (SA) and idiopathic pulmonary fibrosis (IPF) were investigated for alveolar macrophages (AM), secretion of prostaglandin E2 (PGE2) and interleukin 1 (IL-1), together with superoxide anion (O2-) production. Peripheral blood monocytes (PBMO) of the same patients were examined concomitantly for suppressive activity. Consistent with previous results, AM obtained by bronchoalveolar lavage (BAL) from ILD patients markedly suppressed the effects of PHA stimulation of autologous peripheral blood lymphocytes (APL): 61.8 +/- 9.7% suppressive activity compared to 15.5 +/- 15.4% in the control group (CO) P less than 0.001. The AM suppressive activity was correlated with an increase in PGE2 secretion: 3.861 +/- 2.194 ng/10(5) cells/ml in the IPF group, but not in the sarcoid group: 0.217 +/- 0.116 ng/10(5) cells/ml (P less than 0.001 between them). On the other hand, IL-1 secretion by AM was greatly increased in sarcoid patients (308 +/- 196 U/ml) but was within the normal limits in IPF (27.3 +/- 28.8 U/ml, P less than 0.01 between them). Therefore, an inverse correlation was found between degree of PGE2 secretion and IL-1 release by AM in ILD. O2-production by AM was markedly increased in all ILD patients but this mechanism is apparently not involved in suppressive activity. PBMO originating from ILD patients were less suppressive than the corresponding AM. PMID- 2557289 TI - Phagocytosis and bactericidal action of mouse peritoneal macrophages treated with leukotriene B4. AB - The effects of exogenous leukotriene B4 (LTB4) on the resistance of mouse peritoneal macrophages against Salmonella (S.) typhimurium and Pseudomonas (P.) aeruginosa infections were studied. In vitro, LTB4 added to macrophage monolayers at final concentrations of 10(-12)-10(-8) M, enhanced their phagocytosis of S. typhimurium to 2.3 times the control level and that of P. aeruginosa to 1.8 times the control level. The intracellular killing rates were also elevated by the addition of LTB4: for S. typhimurium, 83.3% (LTB4) vs 59.1% (control) and for P. aeruginosa, 46.5% (LTB4) vs 9.2% (control). In vivo, intraperitoneally injected LTB4 (5 ng) enhanced the clearance at 24 h of intraperitoneally injected S. typhimurium from the mouse peritoneal cavity (2.38 x 10(3) +/- 0.94 x 10(3) cells [LTB4] vs 5.73 x 10(5) +/- 1.90 x 10(5) [control]) and spleen (5.00 x 10(2) +/- 0.94 x 10(2) [LTB4] vs 2.47 x 10(4) +/- 0.84 x 10(4) [control]), but this effect disappeared by 48 h. In contrast, in beige mice, an experimental model of the Chediak-Higashi syndrome that is characterized by susceptibility to bacterial infection, there was no induction of the eliminating effect by intraperitoneal injection of LTB4. Activation of macrophages by exogenous LTB4 seemed to have contributed to such an augmented resistance of macrophages to bacterial infection. This study suggested a possible use of LTB4 in bacterial infectious diseases whereby phagocytes are able to play a key role in host defense. PMID- 2557290 TI - Cocaine or delta 9-tetrahydrocannabinol does not affect cellular cytotoxicity in vitro. AB - Cocaine or delta 9-tetrahydrocannabinol (THC), the major psychoactive component of marijuana, has no effect on in vitro cytotoxicity mediated by natural killer (NK) cells and cytotoxic T-lymphocytes (CTL) at concentrations similar to those observed in vivo. PMID- 2557291 TI - Magnetization-transfer n.m.r. investigation of the hydrogen exchange in H2O of the peptide fragment B23-B29 of insulin. AB - Detailed and precise information on the exchanges in water of the peptide hydrogens of the insulin fragment B23-B29 (Gly23-Phe24-Phe25-Tyr26-Thr27-Pro28 Lys29) has been obtained from magnetization-transfer measurements, and nonlinear least-squares fits of the experimental spectra using the expression for the discrete Fourier transform of a sum of exponentially damped sinusoids. From a comparison of the differential exchange rates with those expected for completely solvent-exposed peptide hydrogens, specific conformational features of the heptapeptide are suggested. PMID- 2557292 TI - Clinical and histological responses to a mixed bone/hydroxyapatite graft: report of a case. PMID- 2557294 TI - Determination of vitamin D3 in margarines, oils and other supplemented food products using HPLC. AB - The vitamin D3 content of high fat foods, such as margarines and oils, was determined by liquid chromatography. The method involved sample saponification and an extraction process, in which the purification of the non-saponifiable matter was the critical step. A semi-preparative straight-phase clean up procedure was used to produce an adequately purified sample solution prior to the final analytical quantification step. Vitamin D2 was used as internal standard when estimating the vitamin D3 content. The overall recoveries depended on the fat content of the food sample and varied from 40% for margarines to 80% for low fat products at a concentration of about 10 micrograms/100 g. The minimum detectable amount was 0.36 ng vitamin D3. The mean relative recovery of vitamin D3 in relation to vitamin D2 was calculated to be 0.97. PMID- 2557293 TI - Nutritive utilization of phosphorus in the rat: influence of intestinal resection and dietary medium chain triglycerides and vitamin D3. AB - The effects of intestinal resection and diet on the digestive and metabolic utilization of phosphorus were studied in adult rats from which 50% of the distal small intestine had been removed and in sham-operated controls. Metabolic parameters were measured both 1 and 3 months after surgery. The loss of half of the distal small intestine led to a decline in digestive utilization of phosphorus 1 month after surgery as reflected in bone mineral content. Digestive efficiency had improved by 3 months after surgery. One month's feeding with a diet in which fat was provided as equal parts of medium chain triglycerides, sunflower seed oil and olive oil instead of 100% olive oil enhanced phosphorus absorption and retention, although this improvement was less evident after 3 months. The negative effects of distal small intestine resection on the nutritive utilization of phosphorus were not only palliated but significantly enhanced by supplementing the diet with vitamin D3 at a rate of 0.08 mg/100 g diet. This dose is within physiological limits, and favors phosphorus deposition in bone tissue. PMID- 2557295 TI - Dietary restriction augments superoxide generation in mouse peritoneal macrophages: an investigation using a chemiluminescence probe specific for superoxide anion. AB - It has not been clarified whether dietary restriction alters macrophage functions, although the augmentation of T cell functions by dietary restriction is well known. Forty percent dietary restriction on 9-week-old male C3H/He mice caused a decrease of body weight. However, one of the major macrophage functions, the generation of superoxide anion (O2), was augmented in proteose peptone elicited peritoneal macrophages (MPs) from diet-restricted mice. This increase was more striking when the cells were stimulated by 12-o-tetradecanoylphorbol-13 acetate (TPA), which directly activated protein kinase C, than by opsonized zymosan which binded to receptors on the cells. These results strongly suggest that the augmentation of O2- generation in MPs by dietary restriction is due to the increased activity of protein kinase C which phosphorylate and activate O2 generating enzyme system NADPH oxidase. It is thought that one of the major factors for the reduced incidence of tumor and infection in diet-restricted animals is the augmentation of O2-generation in MPs. PMID- 2557296 TI - Regulation of brain atrial natriuretic peptide and angiotensin receptors: quantitative autoradiographic studies. PMID- 2557297 TI - Nerve blood flow and oxygen delivery in normal, diabetic, and ischemic neuropathy. PMID- 2557298 TI - Activity of batracylin (NSC-320846) against solid tumors of mice. AB - Batracylin (NSC 320846) is a water insoluble, solid tumor active compound discovered by the Development Therapeutics Program of the National Cancer Institute (NCI). In vivo, the NCI found this compound to be highly active [median treated tumor mass/median control tumor mass (T/C) = 0 to 20%] both orally and intraperitoneally against colon 38. In a disk diffusion, soft agar colony formation assay (500 ug/disk), we found solid tumor selectively (compared to leukemia L1210) against colon adenocarcinoma 38 (0-170 zu:L1210 leukemia; greater than 950 zu:C8), colon adenocarcinoma 9 (0-170 zu:L1210; greater than 950 zu:C9), colon adenocarcinoma 7/A (0-170 zu:L1210; 250-400 zu:C7), and pancreas ductal carcinoma 03 (0-170 zu:L1210; greater than 950 zu:Panc 03 (200 zone units [zu] = 6.5 mm zone of inhibition of cultured tumor colonies from drug disk). In vivo we have tested batracylin against mammary adenocarcinoma 16/C, colon 9, colon 38, colon 51, Panc 03, and hepatoma 129. Upon oral administration, batracylin was effective against colon 9 (T/C = 2.4%) and marginally active against colon 38 (T/C = 39%). Batracylin was orally ineffective against Panc 03 (T/C greater than 100%), colon #51 (T/C = 77%) and hepatoma 129 (T/C greater than 100%). Upon subcutaneous administration, batracylin was effective against colon #9 (T/C = 0%), and Panc 03 (T/C = 15%) but ineffective against mammary 16/C (T/C greater than 100%). At efficacious doses, delayed neurotoxicity, hepatic toxicity and a significant host weight loss was noted (with slow recovery). Both our in vitro data and the NCI in vivo data confirm its scant activity against L1210 (%ILS = 8 to 16%). Although showing activity against selected murine solid tumors, it lacked curative potential with early stage disease [C38, C9, Panc 03] and has shown relative inactivity in vitro against human solid tumor cell lines (H-125, CX-1, HCT-8, HCT-116). Batracylin has entered large animal toxicology trials at the NCI, anticipating phase I clinical evaluation. PMID- 2557299 TI - Reduced variation in the clonogenic assay obtained by standardization of the cell culture conditions prior to drug testing on human small cell lung cancer cell lines. AB - An advantage of established tumor cell lines compared to fresh human tumor specimens used in sensitivity assessments is the possibility of repeated experiments. Ultimately a database of sensitivity profiles on a panel of cell lines can be made and the sensitivity to new drugs compared with historical data. A prerequisite of this strategy is a minimal interexperimental variation. The sensitivity of eight human small cell lung cancer cell lines to adriamycin, daunomycin, aclacinomycin A, and mitoxantrone was tested in the clonogenic assay. A covariation in the sensitivity to the drugs emphasized the importance of simultaneous drug testing on the same batch of cells. On one cell line (NCI-N592) the interexperimental variation was further evaluated and a significant correlation was found between preexposure culture conditions, size of S-phase, and sensitivity to adriamycin, daunomycin, and mitoxantrone. Rigorous standardization of the growth conditions prior to clonogenic assay reduced the variation in the sensitivity to adriamycin from a factor of five to only 10-15%. It is concluded that simultaneous experiments on the same batch of cells in drug comparisons should be used if possible. Specification and standardization of culture conditions are necessary in the comparison of drugs tested in different experiments. Inclusion of the same reference drug in all experiments may further increase the validity of comparisons in different experiments. PMID- 2557300 TI - Phase II study of esorubicin (4'-deoxydoxorubicin) in advanced epithelial carcinoma of the ovary: a Gynecologic Oncology Group study. AB - Twenty-six patients with advanced, measurable epithelial carcinoma of the ovary were treated with 76 courses of esorubicin at doses ranging from 20-30 mg/m2 every 3 weeks. All patients are evaluable for toxicity and response. All patients had received prior therapy including radiation therapy in 9, non-anthracycline chemotherapy in 25, and surgery in 25. All patients were GOG performance status 0, 1 or 2. Two partial responses were seen. Severe (grade 3 or 4) leukopenia, thrombocytopenia, and anemia were seen in 13, 1 and 4 patients, respectively. Moderate gastrointestinal toxicity was also noted. Alopecia and mucositis were rare. Phlebitis was not seen. Neither clinical congestive cardiomyopathy nor decrement in left ventricular ejection fraction was observed. We conclude that using this dose and schedule of esorubicin as second-line chemotherapy in ovarian epithelial neoplasms lacks significant activity and is associated with moderate toxicity. PMID- 2557301 TI - Benign synovioma presenting as mechanical abnormality in large lower limb joints. AB - Benign synovioma is a rare cause of symptomatic instability of joints of the lower limb. We report two cases presenting in this way. Arthrotomy is indicated to ensure adequate excision of these lesions if recurrence is to be avoided. PMID- 2557302 TI - Article contributors and topics in practice-oriented health care management journals from 1983 to 1987. AB - This article reports on who is writing articles in the practice-oriented health care management journals and what topics are covered along with the contribution levels of the faculty from the U.S. Health Services Administration education programs. PMID- 2557303 TI - Survival and quality of life after interstitial implantation of removable high activity iodine-125 sources for the treatment of patients with recurrent malignant gliomas. AB - Between January 1980 and January 1988, 95 evaluable patients with recurrent, unifocal, supratentorial malignant gliomas were reirradiated with high-activity iodine-125 sources implanted directly into tumor in afterloaded, removable catheters using computerized tomography-directed stereotaxy. A tumor dose of 5270 15,000 cGy was delivered at a maximum distance of 0.5 cm from the rim of the contrast-enhancing mass seen on CT scans. The median survival for the 50 patients with anaplastic astrocytoma was 81 weeks and for 45 patients with glioblastoma multiforme it was 54 weeks. The 18- and 36-month survival rates for patients with anaplastic astrocytoma were 46% and 28%, respectively; the 18- and 36-month survival rates for patients with glioblastoma multiforme were 22% and 8%, respectively. Because of clinical deterioration, increasing steroid dependency, and increasing mass effect at the implantation site seen on CT scans, necrotic tissue was excised from 47 patients (49%) at craniotomy; in some patients, tumor was mixed with necrotic tissue. The survival of reoperated patients was significantly longer compared with patients who did not undergo this procedure. Serial determination of the Karnofsky Performance Score (KPS) showed that there was no significant deterioration for the group as a whole during the 6 months immediately after implantation. At 18 months, 33 of the patients were alive; KPS ranged between 50 to 90 (mean 79) and 67% were steroid dependent. At 36 months, 18 patients were alive; 17 patients were evaluable with KPS that ranged between 40 to 90 (mean 76) and 53% were steroid dependent. Eleven of the 17 evaluable long-term survivors had a KPS of 80 or higher with a mean of 87. Interstitial brachytherapy may provide long-term survival in selected patients with recurrent malignant gliomas who have been irradiated previously with conventional teletherapy. The quality of life in the majority of long-term survivors appears to be quite satisfactory. Further attempts to control tumor growth using this modality appear to be warranted. PMID- 2557304 TI - Results of combination chemotherapy and thoracic radiation therapy for unresectable non-small cell carcinoma of the lung. AB - From October 1979 to December 1982, 126 patients with locally advanced unresectable or inoperable Stage II (7 patients), Stage IIIA (81 patients), and Stage IIIB (38 patients) non-small cell carcinoma of the lung were treated in a prospective randomized trial using five cycles of CAP (Cytoxan, Adriamycin, and cisplatin), T-CAP (triazinate plus CAP), or V-CAP (VP-16 plus CAP) chemotherapy with thoracic radiation therapy (TRT). TRT consisted of 40 Gy in 10 fractions (split-course) with cycles 3 and 4 of chemotherapy. The treatment field included the primary tumor, ipsilateral hilum, mediastinum, and ipsilateral supraclavicular fossa. All patients were followed until death or for a minimum of 5 years for survivors. The evaluable subgroup consisted of 102 patients who completed TRT. Median and 5-year survivals for the entire group were 14.0 months and 10%, respectively; for the evaluable subgroup, they were 14.8 months and 12%, respectively. There was a trend toward better survival with V-CAP plus TRT than with CAP plus TRT (p = 0.08). Median and 5-year survivals were 16.2 months and 18%, respectively, with V-CAP plus TRT. Of eight prognostic variables analyzed for their association with survival, only Eastern Cooperative Oncology Group performance status (0,1 versus 2) (p = 0.02) and weight loss (less than or equal to 10% versus greater than 10%) (p = 0.05) were significant. Sex, age, T stage, N stage, overall stage, and histologic type were not significantly associated with survival. Failure analysis revealed 83 patients (81%) with identifiable first failures. The median time to first failure was 9.8 months, and the median survival after first failure was 4.7 months. Failure patterns included local failure alone (19%), local and distant (20%), and distant alone (43%). Nineteen percent of patients had no documented progression. Total failure patterns were local in 39% and distant in 63%. Twenty-three patients (23%) had failure in the brain; they accounted for 31% of all distant failures. In 20 of these patients (20% of all patients), this was the only site of failure. There were eight (8%) initial nodal failures in 96 untreated contralateral supraclavicular fossae. No initial failures were seen in any of 101 untreated contralateral hila. The data suggest the following: (a) Combined treatment with V-CAP and TRT yielded excellent results (median survival, 16.2 months; 5-year survival, 18%).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557305 TI - Breast conserving surgery and definitive irradiation for early stage breast cancer. AB - Breast conserving surgery and postoperative breast radiotherapy were used to treat 219 cases of AJCC Stage I and II breast carcinoma at the Michael Reese and University of Chicago Hospitals. Most patients were treated with lumpectomy and axillary sampling followed by breast irradiation to a dose of 46 Gy followed by a boost dose of 14-16 Gy to the surgical bed. The 5-year actuarial local control is 92%. Follow-up is 1 to 10 years and the median follow-up is 36 months. Of the seven patients who recurred in the breast, three failed in the boost site and three failed adjacent to the boost site. The seventh patient recurred diffusely in the breast and skin. Four of the seven recurrences were in patients with positive surgical margins. The 5-year actuarial relapse-free survival is 80%. Factors which had an adverse affect on the cosmetic results were a scar length greater than 8 cm and a volume of resected breast tissue greater than 100 cm3. Treatment related complications were minor and infrequent. Breast conserving surgery followed by radiation therapy is effective in achieving local control with good to excellent cosmetic results. PMID- 2557306 TI - Can the clinical and mammographic findings at presentation predict the presence of an extensive intraductal component in early stage breast cancer? AB - The presence or absence of an extensive intraductal component (EIC) in early stage infiltrating ductal breast carcinoma has been considered to be an important factor in determining the extent of breast resection required prior to radiation therapy. It would therefore be useful if the presence or absence of an extensive intraductal component in a breast tumor could be predicted pre-operatively. To determine whether selected radiographic features might be correlated with whether or not a cancer is EIC+, we reviewed the pre-operative mammographic findings in 105 cases of Stage I and II infiltrating ductal carcinoma. Forty-one cases were EIC+ and 64 were EIC-. Both EIC+ and EIC- tumors were commonly detectable by mammography (93% and 83%, respectively, p = NS). EIC+ cancers, however, were significantly more likely to show microcalcifications with or without a mass compared to EIC- cases (83% vs 27%, p less than 0.0001). In particular, the presence of microcalcifications without a mammographic mass was more common for EIC+ cancers than EIC- cancers (34% vs 5% p = 0.0002). Conversely, a soft tissue mass without microcalcifications was seen mammographically in 56% of EIC- cases, compared to only 10% of EIC+ cases (p less than 0.0001). Predictive value calculations showed that the presence of microcalcifications in the absence of a mammographic mass conveys a 73% likelihood a cancer will be EIC+ (95% confidence interval = 39-94%). The positive predictive value of a mammographic mass or architectural distortion without microcalcifications for an EIC- cancer was 92% (95% confidence interval = 79-98%). We conclude that the mammographic findings may be useful pre-operatively in differentiating between patients with and without an EIC. Microcalcifications are much more commonly associated with EIC+ cancers than EIC- cancers, and the presence of an EIC- cancer without a mammographic mass is infrequent. Further characterization of the extent and pattern of microcalcifications might improve the predictive value of mammography in the pre-operative identification of patients with an EIC. PMID- 2557308 TI - Failure of accelerated neutron therapy to control high grade astrocytomas. AB - Sixty-two patients with high grade malignant astrocytoma were treated with fast neutrons using three different treatment schemes to evaluate the effect of shortening the overall time. Dose and fraction number were kept constant. The total dose was 16-18 neutron Gy delivered in six fractions, weekly for 6 weeks, twice a week over 3-4 weeks, or three times a week over 2 weeks. There were no obvious differences in survival times among the three groups. We conclude that accelerated neutron therapy does not improve survival of patients with grade 3 and 4 astrocytoma. PMID- 2557307 TI - 194 hepatocellular cancers treated by radiation and chemotherapy combinations: toxicity and response: a Radiation Therapy Oncology Group Study. AB - Hepatocellular carcinoma is known to have a doubling time of approximately 41 days. This rapid cell division suggested that hyperfractionated radiation and chemotherapy might add an advantage in gaining remission of this malignancy. One hundred and thirty-five patients (70% with metastasis and/or previous treatment) were prospectively treated with single daily fractions to the liver (3.0 Gy external beam radiation, total dose 21.0 Gy), and chemotherapy for hepatocellular carcinoma. The low dose chemotherapy used in conjunction with the radiation was 2 hr before treatment on days 1, 3, 5, and 7 and consisted of Adriamycin, 15 mg IV and 5-FU, 500 mg IV. These patients were compared to a second group of 59 patients (80% with metastases and/or previous treatment) treated using the same chemotherapy regimen but using hyperfractionated whole liver external beam irradiation (1.2 Gy twice daily, 4 hr between treatments, 5 days per week to 24.0 Gy, 10 MV photons). Response was determined by CT scan tumor volumetric analysis. The response rate for the single daily fraction patient group was 22% and for the new hyperfractionated group, 18% (p = 0.68). Toxicity was evaluated by RTOG criteria. The grade 4 hematologic toxicity noted in the daily fraction patient group was 6%. Among 59 patients treated with the hyperfractionated liver irradiation, 2% experienced grade 4 hematologic toxicity. Esophagitis occurred in 1% of patients in the standard fractionation group and 19% in the hyperfractionated group (p = 0.0001). Grade 1-4 thrombocytopenia occurred in 49% of patients in the conventional group and 68% in the hyperfractionated group (p = 0.03). Normal liver volume changes with treatment were measured with CT scan tumor volumetric analysis. The hyperfractionated group experienced a median of 11 cc increase in liver volume and the conventional group a 46 cc decrease, but the difference was not significant. Hyperfractionated radiation did not demonstrate a significant benefit over standard daily radiation, but acute toxicity appeared to be higher. PMID- 2557309 TI - Twenty year experience with radiation therapy for temporal bone chemodectomas. AB - Between 1965 and 1984, 20 patients with chemodectomas of the temporal bone were seen at The Methodist Hospital in Houston, Texas and at the Cancer Therapy and Research Center in San Antonio, Texas, Ten patients were treated with radiation therapy alone, seven with surgery and post-operative radiation, one with pre operative radiation, and two with radiation therapy following surgical recurrence. Most patients had advanced tumors at presentation. Radiation doses ranged from 22.5 Gy to 50.0 Gy. The most frequent dose was 45.0 Gy, given in 225 cGy fractions, 9.0 Gy per week. The most common radiation portal arrangement was oblique fields with paired wedges. There were no local failures or significant radiation induced complications among the patients with benign chemodectomas. The follow-up period ranged from 3 to 23 years (mean 11 years). Only one patient developed systemic metastases and progression of the primary temporal bone chemodectoma. These results and a review of the literature demonstrate that radiation therapy alone is a safe and effective treatment modality for chemodectomas of the temporal bone. PMID- 2557310 TI - Radiation therapy treatment planning in supratentorial glioblastoma multiforme: an analysis based on post mortem topographic anatomy with CT correlations. AB - In a previous study of the brains of 15 adults with glioblastoma multiforme who received minimal or no radiotherapy we determined the topographic distribution of tumor cells. All 15 brains had been fixed and then cut in the coronal or horizontal plane. The distribution of neoplastic cells was determined and entered onto tracings of the whole mount histologic sections. The last CT scans obtained prior to death of 11 of the patients were reviewed independently by a neuroradiologist who traced, on the CT scans, the outer edge of both the contrast enhancing area and the peritumoral low density "edema". Presented with the neuroradiologist's assessment of the contrast enhancing rim of tumor and of the "edema", a radiotherapist and a radiation dosimetrist, in the present study, prepared treatment plans for a 6 MeV linear accelerator. In 9 of the 11 cases in which immediately antemortum CT scans were available, radiation treatment of the contrast enhancing area alone with a 1 cm margin would have missed portions of the histologically identified tumor. Treatment of the contrast enhancing area along with the peritumoral "edema", with a 1 cm margin, would have covered histologically identified tumor in six of the 11 cases. Treatment of the contrast enhancing area, all "edema", and a 3 cm margin around the "edema" would have covered histologically identified tumor in all cases. Tumors tended to track along nerve pathways. In those lesions near the midline it was common for tumor to cross the corpus callosum. We conclude that radiotherapy with fields designed to treat the contrast enhancing region alone or this region plus "edema" with a tight margin will frequently miss tumor which can be histologically identified by our technique. PMID- 2557311 TI - Serologic status of pseudorabies virus in growing-finishing pigs in quarantined herds. AB - It has been reported that pseudorabies virus (PRV) stops spreading within growing finishing sections of a large percentage of infected farrow-to-finish herds. This study was designed to follow the PRV status of growing-finishing pigs in a sample of infected herds. Fifteen infected herds were selected, of which 11 had seropositive finishing pigs and 4 had seronegative finishing pigs. These herds were visited quarterly for one year, and a cross section of growing-finishing pigs was tested for the presence of anti-PRV antibodies. The 4 herds that initially were seronegative remained seronegative, whereas of the 11 herds initially seropositive, 4 remained seropositive, 4 became seronegative, and 3 became temporarily seronegative before becoming seropositive again. Three characteristics serologic profiles were observed: one indicating continued viral spread; one indicating no spread for at least the preceding 3 months; and one indicating that PRV spread had recently ceased in this section of the herd. Results of our study indicated that periodic monitoring of a cross section of the growing-finishing pigs for their PRV serologic status was valuable for determining whether PRV was actively spreading in this section of the herd. PMID- 2557312 TI - On interpretation of feline leukemia test results. PMID- 2557313 TI - Identification of pseudorabies virus-infected swine herds by evaluating the serostatus of boars or finishing pigs. AB - Data were collected from 39 Minnesota swine farms quarantined for pseudorabies virus (PRV) infection. Each herd was serologically evaluated for antibodies to PRV in the sows, boars, and finishing pigs. To identify PRV-seropositive swine herds, the Kappa statistic was used to estimate the effectiveness of evaluating the PRV serostatus of boars or of finishing pigs. Using the serostatus of all herd boars, the sensitivity (with 95% confidence interval) of identifying PRV infected herds was 58 +/- 22%, and the specificity was 100 +/- 0%; Kappa statistic was 0.55. Using the serostatus of a representative sample of finishing pigs, the sensitivity of identifying PRV-infected herds (with 95% confidence interval) was 63 +/- 22%, and specificity was 87 +/- 23%; Kappa statistic was 0.40. The PRV serostatus of herd boars or of a representative sample of finishing pigs did not accurately reflect the PRV serostatus of the herd. PMID- 2557314 TI - Lack of evidence of transmission of bovine leukemia virus by rectal palpation of dairy cows. AB - A study was undertaken in a California dairy to estimate the risk of transmission of bovine leukemia virus associated with rectal palpation of cows. The probability of seroconversion within 3 months after rectal palpation was examined as a function of the prevalence of infection in cows at the time of palpation. It was hypothesized that the probability of seroconversion within 3 months after palpation would be higher for cows in a group with high prevalence of infection than for cows in a group with low prevalence. The probability of seroconversion after rectal palpation was equal to 0.034, and results of logistic regression, using data from 1,116 noninfected and 1,047 infected cows, did not indicate any significant association between probability of seroconversion after rectal palpation and prevalence of infection (P = 0.189). Results of this study suggest that risk of bovine leukemia virus transmission by rectal palpation typically used in reproductive tract examination of cows either does not occur or is uncommon. PMID- 2557315 TI - Coexpression of platelet-derived growth factor (PDGF) A-chain and PDGF receptor genes in human gastric carcinomas. AB - In this study we examined the expression of platelet-derived growth factor (PDGF) A-chain and PDGF receptor genes in seven human gastric carcinoma cell lines and 15 gastric carcinoma tissues. Expression of mRNA for PDGF A-chain was found in all gastric cell lines and all gastric carcinoma tissues. Two of the seven gastric carcinoma cell lines expressed PDGF receptor mRNA. Out of the 15 gastric carcinoma tissues, eight showed enhanced expression of PDGF receptor mRNA and all of them demonstrated prominent fibrous stroma. Moreover, the incidence of enhanced expression of PDGF receptor mRNA was higher in scirrhous carcinoma than in well differentiated adenocarcinoma. These results strongly suggest that PDGF produced by tumor cells acts as a paracrine growth factor for production of fibrous stroma in gastric carcinomas. PMID- 2557317 TI - Synergistic interaction of josamycin with human neutrophils bactericidal function in vitro. AB - Josamycin, a 16-membered ring macrolide is concentrated up to 20-fold in phagocytic cells compared with serum. We have studied the in-vitro interaction of this drug with human neutrophils (PMN) bactericidal function by using two strains resistant to this antibiotic, Pseudomonas aeruginosa and Klebsiella pneumoniae, and a sensitive one, Staphylococcus aureus 209P. It was shown that josamycin pretreated adherent PMN displayed an increased phagocytic activity (about 30 to 40%) for S. aureus or K. pneumoniae, mainly due to the recruitment of an additional phagocytizing subset of PMN. Furthermore, the bacterial killing was enhanced in josamycin-treated PMN in a dose-dependent manner for K. pneumoniae (60-130% increase in the range of concentration 0.1-25 mg/l) and independently of the dose for S. aureus (about 425-460% increase for josamycin 0.1-10 mg/l). P. aeruginosa killing by whole blood was also significantly increased in the presence of 10 and 1 mg/l of josamycin. Other PMN functions were not much altered by josamycin except an enhancement of the formyl-methionyl-leucyl-phenylalanine induced oxidative response. Chemotaxis was only increased by the presence of a high concentration (100 mg/l) of josamycin. These data suggest that the bactericidal synergy between PMN and josamycin could be related, partly at least, to a direct enhancing effect of josamycin on some PMN functions such as phagocytosis, chemotaxis and FMLP-induced chemiluminescence. On the other hand, alterations of bacteria, either inside the phagolysosome or in the extracellular medium, could lead to an enhanced susceptibility to the phagocytes' microbial mechanisms. PMID- 2557318 TI - In-vitro effects of cilofungin (LY121019), amphotericin B and amphotericin B deoxycholate on human polymorphonuclear leucocytes. AB - The in-vitro influence of cilofungin (LY121019) and amphotericin B on human polymorphonuclear leucocytes (PMNs) was studied by a multifunctional approach. Cilofungin at high concentration (greater than or equal to 20 mg/l) increased adherence to plastic and ingestion of Staphylococcus aureus by PMNs in suspension and Candida albicans by adherent PMNs, and slightly decreased MTT reduction and superoxide generation. Amphotericin B and amphotericin B-deoxycholate decreased adherence to plastic (IC50: 5.1 and 8.2 mg/l respectively) and superoxide generation induced by PMA and opsonized zymosan (IC50 1.1 mg/l for amphotericin B deoxycholate). Variable results were observed for intra-cellular killing and ingestion. The functional assessment was made with four clinical isolates of yeasts (Can. albicans, Can. tropicalis, Can. Torulopsis) (glabrata, Cryptococcus neoformans). The inoculum was preincubated with the antifungals (PRE) or they were added during (PER) or after ingestion (POST) using PMNs in suspension (PRE and PER), or adhering to plastic (PRE, PER and POST). With the scheme PRE, killing was usually increased with amphotericin B-deoxycholate and cilofungin. With Crypt. neoformans, ingestion was also increased by the antifungals and sodium-deoxycholate, probably by altering the capsule. The results of the scheme POST showed that amphotericin B-deoxycholate, but not cilofungin, increased intracellular killing of Can. albicans, Can. tropicalis and Can. glabrata. PMID- 2557316 TI - Binding properties and proliferative effects of human recombinant granulocyte macrophage colony-stimulating factor in primary leukemia and lymphoma. AB - Binding of radiolabeled human granulocyte-macrophage colony-stimulating factor (GM-CSF) was studied with blast cells from eight patients with acute myeloblastic leukemia (AML), and neoplastic lymphoid cells from one patient with acute lymphoblastic leukemia (ALL), two patients with chronic lymphocytic leukemia (CLL) and one patient with undiagnosed B cell neoplasia. In all AML cases studied, Scatchard graphs of the direct binding data were curvilinear, and were best fitted by curves derived from a two-binding-site model; one site with high affinity (Kd1 = 12-71 pM; 174-602 sites/cell) and the other with low affinity (Kd2 = 0.5-2.7 nM; 1137-6020 sites/cell). A cross-linking study on blast cells from one AML patient demonstrated specific bands which were similar to those reported for peripheral blood neutrophils. Furthermore, blast colony assays for the same preparations showed remarkable proliferative response to GM-CSF in the concentration range from 0.3 nM to 7.0 nM (ED50 greater than 0.7 nM). This concentration range is approximately one order of magnitude higher than that which is effective for colony formation from normal bone marrow progenitors (ED50 in equilibrium 0.1 nM). No significant correlation could be observed between the responsiveness of blast progenitors to GM-CSF, and the numbers or affinities of GM-CSF binding sites demonstrated on blast cells. In studies with neoplastic lymphoid cells from four patients, 125I-GM-CSF also specifically bound in two cases, while response to GM-CSF was not observed in these cases. These results indicate that the expression of GM-CSF receptor is not restricted to the GM-CSF responsive AML blast cells, but can be observed in other AML blast cells and even in neoplastic lymphoid cells. PMID- 2557319 TI - Absence of significant interaction of fluconazole with cyclosporin. AB - The effect of a new azole antifungal drug, fluconazole, on cyclosporin concentration and pharmacokinetics was studied in ten bone marrow transplant recipients. There were no significant statistical or clinical differences in mean cyclosporin trough concentrations, peak concentrations and AUCs after fluconazole given either as a single 100 mg oral dose or as 100 mg daily for 14 days. Fluconazole was well tolerated without drug related side effects. Slight to moderate increases of liver laboratory parameters were observed in four patients during the multiple dose sequence, but these changes might be attributable, in part, to the underlying diseases or concomitant therapies. The data suggest a lack of clinically relevant pharmacokinetic interaction of fluconazole with cyclosporin. It is concluded that fluconazole may be administered to cyclosporin treated patients without enhanced risk of toxicity. PMID- 2557320 TI - Muscle morphological and biochemical adaptations to training in obese Zucker rats. AB - The purposes of the present study were to characterize the histochemical and enzymatic profiles of various hindlimb skeletal muscles, as well as to determine maximal O2 consumption (VO2max) and respiratory exchange ratios (R) during steady state exercise in the obese Zucker rat. The changes that occurred in these parameters in response to a 6-wk training program were then assessed. Obese rats were randomly assigned to a sedentary or training group. Lean littermates served as a second control. Training consisted of treadmill running at 18 m/min up an 8% grade, 1.5 h/day, 5 day/wk for 6 wk. During week 6, VO2max and R during a steady state run (74% max) were determined. After 2 days of inactivity, hindlimb muscles were excised, stained for fiber type and capillaries, and assayed for hexokinase, citrate synthase, cytochrome oxidase, and beta-hydroxyacetyl-CoA dehydrogenase. The obese sedentary rats demonstrated greater oxidative enzyme activities per gram of muscle tissue than their lean littermates, greater R values during submaximal exercise of the same relative intensity, and greater absolute VO2max values. Training resulted in a 20-56% increase in oxidative enzymes, a 10% increase in VO2max, and an increase in capillary density in the soleus and plantaris. There was no alteration in R values during exercise at 74% VO2max or in fiber type composition in response to exercise training. Results suggest that the muscle of the obese Zucker rat manifests a greater oxidative capacity than the muscle of its lean littermates. The apparent inability of the obese rat to increase its use of fat during submaximal exercise of the same relative intensity in response to training remains to be elucidated. PMID- 2557321 TI - Impulse propagation and muscle activation in long maximal voluntary contractions. AB - With fatigue, force generation may be limited by several factors, including impaired impulse transmission and/or reduced motor drive. In 5-min isometric maximal voluntary contraction, no decline was seen in the peak amplitude of the tibialis anterior compound muscle mass action potential (M wave) either during or immediately after the voluntary effort, provided maximal nerve stimulation was retained. For first dorsal interosseous (FDI) muscle, M wave amplitudes declined by 19.4 +/- 1.6% during the first 2 min but did not change significantly thereafter, despite the continued force reduction (up to 94% in 5 min for both muscles). The duration of the FDI M waves increased (greater than 30%), suggesting that the small decline in amplitude was the result of increased dispersion between the responses of different motor units. Some subjects kept FDI maximally activated throughout, but when they used tibialis anterior, twitch occlusion and tetanic muscle stimulation showed that most subjects were usually only able to do so for the first 60 s and thereafter only during brief "extra efforts." Thus force loss during isometric voluntary contractions sustained at the highest intensities results mainly from failure of processes within the muscle fibers. PMID- 2557322 TI - Effect of induced alkalosis on perception of exertion during intermittent exercise. AB - The purpose of this study was to evaluate the effects of metabolic alkalosis on differentiated ratings of perceived exertion during intermittent high-intensity exercise. Six endurance-trained females participated as subjects in this investigation. Each subject underwent three separate experimental trials in which NaHCO3 was ingested in either a single (0.3 g NaHCO3/kg body wt) or periodic schedule (0.12 g NaHCO3/kg body wt initially, with 0.18 g/kg body wt distributed in equal doses before each 5-min exercise bout). Calcium carbonate served as a placebo control. An intermittent exercise protocol was used in which each subject rode a cycle ergometer at 90% maximum O2 consumption for 5 min. Within each acid base condition, the exercise protocol was repeated three times with 10-min rest periods interspersed. Differentiated ratings of perceived exertion for the legs (RPE-L), chest (RPE-C), and overall body (RPE-O) were attenuated under alkalotic treatment relative to placebo control regardless of pattern of NaHCO3 administration. RPE-L, RPE-C, and RPE-O were negatively correlated to the bicarbonate concentration of venous blood. This investigation suggests that perception of effort during high-intensity intermittent exercise can be related to buffering capacity of the blood. PMID- 2557323 TI - Skeletal muscle beta-receptors and isoproterenol-stimulated vasodilation in canine heart failure. AB - To investigate whether heart failure alters beta-adrenergic receptors on skeletal muscle and its associated vasculature, the density of beta-adrenergic receptors, isoproterenol-stimulated adenylate cyclase activity, and coupling of the guanine nucleotide-binding regulatory protein were compared in 18 control dogs and 16 dogs with heart failure induced by 5-8 wk of ventricular pacing at 260 beats/min. Hindlimb vascular responses to isoproterenol were compared in eight controls and eight of the dogs with heart failure. In dogs with heart failure, the density of beta-receptors on skeletal muscle was reduced in both gastrocnemius (control: 50 +/- 5; heart failure: 33 +/- 8 fmol/mg of protein) and semitendinosus muscle (control: 43 +/- 9; heart failure: 27 +/- 9 fmol/mg of protein, both P less than 0.05). Receptor coupling to the ternary complex, as determined by isoproterenol competition curves with and without guanosine 5'-triphosphate (GTP), was unchanged. Isoproterenol-stimulated adenylate cyclase activity was significantly decreased in semitendinosus muscle (control: 52.4 +/- 4.6; heart failure: 36.5 +/ 9.5 pmol.mg-1.min-1; P less than 0.05) and tended to be decreased in gastrocnemius muscle (control: 40.1 +/- 8.5; heart failure: 33.5 +/- 4.5 pmol.mg 1.min-1; P = NS). Isoproterenol-induced hindlimb vasodilation was not significantly different in controls and in dogs with heart failure. These findings suggest that heart failure causes downregulation of skeletal muscle beta adrenergic receptors, probably due to receptor exposure to elevated catecholamine levels, but does not reduce beta-receptor-mediated vasodilation in muscle. PMID- 2557325 TI - Oxidation/reduction state of cytochrome oxidase during repetitive contractions. AB - There is disagreement regarding whether inadequate O2 determines maximal O2 uptake (VO2max) and lactic acid output (L) during muscular activity. Direct assessment of mitochondrial cytochrome oxidase (cytochrome a-a3) oxidation/reduction (O/R) state should provide an unequivocal answer for this issue. A new near-infrared spectrophotometric method was used to measure the O/R state of cytochrome a-a3 of dog gastrocnemius-plantaris muscle in situ during repetitive isotonic twitch and tetanic contractions. Three contraction frequencies were used for each contraction type in alternating sequence to provide a wide range of VO2 up to VO2max. VO2 and L were measured after 3 and 9 min of a 10-min contraction period, and 15 min were allowed for recovery between contraction periods. VO2 increased with contraction frequency. L was variably increased with contraction frequency at 3 min and uptake usually occurred at 9 min, except at the highest tetanic frequency. The O/R span of cytochrome a-a3 was determined by respiring the animals with 100% N2 to determine the most reduced state. This was followed by respiration with 100% O2, which gave the most oxidized state transiently during recovery. Within this span in muscles at rest, cytochrome a-a3 was 50-80% oxidized. During contractions of both types at all frequencies, cytochrome a-a3 always became more oxidized by an additional 10-20%. These findings should put to rest any arguments that inadequate O2 is a determinant of VO2max or L under the conditions of these experiments: repetitive contractions with free flow in self-perfused muscles and normoxia. PMID- 2557324 TI - Effects of ovariectomy on energy metabolism in exercising rat muscle studied by 31P-NMR. AB - The effects of ovariectomy on metabolism of high-energy phosphate compounds during and after exercise were studied in hindleg muscles of 14 rats. Sciatic nerve stimulation was used to establish different work loads, and the changes in inorganic phosphate-to-phosphocreatine ratios (Pi/PCr) were recorded by 31P nuclear magnetic resonance (NMR) in vivo. Four weeks after ovariectomy, there was evidence of significantly higher Pi/PCr during work at stimulation rates greater than 0.5 Hz. The slope for the stimulation rate-to-Pi/PCr relationship decreased from 1.98 +/- 0.15 to 1.36 +/- 0.2 Hz/Pi/PCr after ovariectomy. The normalized tension output of these muscles, tested separately using identical stimulation protocols, was not changed with ovariectomy. Thus the relationship between work (tension-time integral) and bioenergetic cost (Pi/PCr) suggested reduced maximal enzyme activity (Vmax) by 9-17% as a result of lack of ovarian sex hormones, but no change in Michaelis-Menten constant (Km) was found. Postexercise recovery was also significantly slower (3.27 +/- 0.54 PCr/Pi units per minute compared with 4.04 +/- 1.08 in controls). It is suggested that reduced levels of ovarian sex hormones decrease oxidative phosphorylation. Cytochrome oxidase activity was reduced in these muscles by 40%, but other mitochondrial enzyme systems may be affected as well. The possible significance of these data is the implication of a reduced capacity for menopausal women or amenorrheic female athletes to perform prolonged intensive exercise. PMID- 2557326 TI - Protein phosphatases come of age. PMID- 2557327 TI - Identification of an alternate type I insulin-like growth factor receptor beta subunit mRNA transcript. AB - Several studies have suggested that heterogeneity exists in the type I insulin like growth factor (IGF) receptor beta subunit. We have examined type I IGF receptor mRNA transcripts by ribonuclease (RNase) protection assay to determine if the heterogeneity could result from alternative splicing of the gene. An area that corresponded to the nucleotide sequence just upstream of the region encoding the transmembrane domain of the beta subunit was identified as being a potential site of alteration in the transcript. Since the 5' and 3' ends were known, polymerase chain reaction was used to clone a cDNA that included this region. Analysis revealed that an alternate type I IGF receptor mRNA transcript with a 3 base pair deletion could account for the results of the RNase protection assay. The deletion changes the amino acid sequence at position 899 substituting Arg for a Thr-Gly. Furthermore, this alternate transcript was ubiquitously found in tissue and cell line RNAs. Although the identified transcript cannot fully account for the documented heterogeneity in type I IGF receptor beta subunit sizes, the results suggest that a form of the beta subunit with an alternate primary sequence may exist. PMID- 2557328 TI - A human membrane-associated folate binding protein is anchored by a glycosyl phosphatidylinositol tail. AB - Membrane-associated and soluble forms of folate binding protein (FBP) have been identified in mammalian tissues and biological fluids. Despite their solubility differences, these two forms are functionally similar, immunologically cross reacting, and have the same apparent molecular weights. In this study we demonstrate, for the first time, that the membrane FBP of cultured human KB cells contains a glycosyl-phosphatidylinositol (GPI) tail which is responsible for its hydrophobic properties and distinguishes it from the soluble FBP released into the medium. Treatment of the purified membrane FBP with phospholipase C specific for phosphatidylinositol (PI-PLC) removed the GPI tail and converted it to the soluble form without a change in apparent Mr by sodium dodecyl sulfate polyacrylamide gel electrophoresis. In addition, virtually all of the folate binding sites on the plasma membrane of the intact cells were released as soluble, functional FBP following treatment with PI-PLC. The GPI tail contained 1 O-alkyl-2-O-acylglycerol as a mixture of fatty alcohols in ether linkage at C1 of the glycerol backbone and almost exclusively docosanoic acid (22:0) as the fatty acid on C2. The inositol also contained a mixture of fatty acids (16:0, 18:0, 18:1, 20:4, 22:0) located on a site other than the C2 position since the FBP was susceptible to PI-PLC cleavage. After nitrous acid deamination, the aqueous portion of the FBP contained covalently bound fatty acids, predominantly palmitate (16:0) and stearate (18:0), indicating the presence of additional acyl groups attached to the peptide in the form of amide, ester, or thioester linkage. PMID- 2557329 TI - Assembly of insulin/insulin-like growth factor-1 hybrid receptors in vitro. AB - Insulin and Mn/MgATP treatment of immunoaffinity-purified alpha beta heterodimeric insulin receptors induced the formation of an alpha 2 beta 2 heterotetrameric insulin receptor complex. In contrast, insulin-like growth factor-1 (IGF-1) treatment was completely ineffective in inducing the association of alpha beta heterodimeric insulin receptors. Similarly, IGF-1 or Mn/MgATP, but not insulin, treatment of immunoaffinity-purified alpha beta heterodimeric IGF-1 receptors induced the formation of an alpha 2 beta 2 heterotetrameric IGF-1 receptor complex. A monoclonal antibody specific for the insulin receptor (MA5) completely immunoprecipitated all the insulin binding activity from both the alpha 2 beta 2 heterotetrameric and alpha beta heterodimeric insulin receptor complexes but did not immunoprecipitate IGF-1 receptors. Conversely, the IGF-1 receptor-specific monoclonal antibody (alpha IR-3) immunoprecipitated all the IGF 1 binding activity, but not insulin receptors. The simultaneous treatment of pooled equal amounts of alpha beta heterodimeric insulin and IGF-1 receptors with a combination of insulin and IGF-1 resulted in the formation of alpha 2 beta 2 heterotetrameric insulin and IGF-1 receptor complexes. However, in the mixed alpha 2 beta 2 heterotetrameric receptor fraction MA5 immunoprecipitated 94% of the insulin binding in addition to 27% of the IGF-1 binding activity whereas alpha IR-3 immunoprecipitated 97% of the IGF-1 binding in addition to 38% of the insulin binding activity. Treatment of the mixed alpha beta heterodimeric insulin and IGF-1 receptors with Mn/MgATP also resulted in the formation of cross immunoreactive (42-46%) alpha 2 beta 2 heterotetrameric receptors. These data directly demonstrate the formation of insulin/IGF-1 hybrid receptors by both a combination of insulin plus IGF-1 or Mn/MgATP treatment of purified human placenta alpha beta heterodimeric insulin and IGF-1 half-receptors in vitro. PMID- 2557330 TI - Mitochondrial electron transport-linked tocopheroxyl radical reduction. AB - alpha-Tocopherol (vitamin E) is a lipophilic chain-breaking antioxidant which inhibits lipid peroxidation in isolated mitochondrial membranes and protects membranes from oxidative damage. The primary oxidation product of vitamin E is the tocopheroxyl radical. Reduction of the tocopheroxyl radical can occur by reactions with water-soluble anti-oxidants such as ascorbate or glutathione, resulting in the recycling of vitamin E. Physiological concentrations of vitamin E are too low to allow detection of tocopheroxyl radical by ESR. After dietary supplementation with vitamin E, a 10-20-fold increase in the rat liver mitochondrial membrane content of vitamin E was achieved and this allowed for direct detection of the tocopheroxyl radical by ESR, after treatment with an oxidizing system composed of lipoxygenase and arachidonic acid. By using submitochondrial particle membranes, it was shown that NADH, succinate, and reduced cytochrome c-linked oxidation reduce the tocopheroxyl radical, preventing both accumulation of the radical and vitamin E consumption. As the electron transport chain can reduce tocopheroxyl radical it may have an important physiological role in recycling vitamin E. PMID- 2557331 TI - Transforming growth factor beta treatment decreases ACTH receptors on ovine adrenocortical cells. AB - Transforming growth factor beta (TGF beta) is a potent inhibitor of adrenocortical cell differentiated functions, whereas corticotropin (ACTH) is the main physiological hormone which acts positively on these functions. We have studied the effects of both TGF beta and ACTH on ovine adrenocortical cell ACTH receptors. Ovine adrenocortical cells contained specific high affinity (Kd = 2.7 +/- 1.6 x 10(-10) M) and low capacity (1190 +/- 120 sites/cell) ACTH receptors. Pretreatment of cells with TGF beta resulted in a time- and dose-dependent (ED50 = 50 pg/ml) decrease of 125I-ACTH1-39 binding. The observed decrease in ACTH binding was due to a 2-3-fold decrease in the number of binding sites without modification of the binding affinity. On the contrary, pretreatment of cells with ACTH caused a 4-4.5-fold increase in the number of ACTH binding sites without an effect on the Kd. When cells were pretreated with both ACTH and TGF beta, TGF beta blocked completely the positive trophic effect of ACTH on its own receptors. The variations in ACTH receptor number were associated with parallel changes on acute ACTH-induced cyclic AMP production. Thus, the effects of TGF beta on ACTH receptor content are likely another important negative action of this peptide on adrenocortical cell differentiation. Moreover, these results suggest that regulation of ACTH receptor number may be one mechanism by which hormones and growth factors control adrenocortical differentiation. PMID- 2557332 TI - The type I insulin-like growth factor receptor is a motility receptor in human melanoma cells. AB - Insulin-like growth factors I and II (IGF-I and II) and insulin are chemotactic agents for the human melanoma cell line A2058. As shown in this report, the motility receptor mediating this response is the heterodimeric type I IGF receptor. These three factors are able to compete with 125I-labeled IGF-I for binding to the cell surface with IC50 values equal to approximately 2 (IGF-I), approximately 150 (IGF-II), and approximately 300 nM (insulin). Cross-linking of 125I-IGF-I to the cell surface with disuccinimidyl suberate followed by analysis with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography reveals a 130-kDa protein (reduced) consistent with the alpha component of a type I receptor and a 38-kDa protein which does not bind insulin, and thus could be another IGF-I cell surface binding protein. The anti-IGF-I receptor monoclonal antibody (alpha IR-3) also competes with labeled IGF-I in binding experiments. In contrast, a control monoclonal antibody, matched to alpha IR-3 with respect to IgG subclass, has no significant effect on IGF-I binding. While alpha IR-3 inhibits the motility induced by IGF-I, IGF-II, and insulin, pertussis toxin (0.01-1.0 micrograms/ml) has no significant effect on the motility induced by the insulin-like growth factors or insulin on this cell line. Therefore, the type I IGF receptor appears to mediate a highly potent pertussis toxin-insensitive motility response to IGF-I, IGF-II, and insulin. In contrast, motility induced by the autocrine motility factor, a cytokine produced by the A2058 cells, is not affected by alpha IR-3 but is extremely sensitive to pertussis toxin. When mixtures of autocrine motility factor and IGF-I are employed to induce chemotaxis, the resulting motility is greater than that induced by either agent alone. These data indicate that motility in this melanoma cell line can be initiated through multiple receptors that stimulate the cells by separate transduction pathways. This capability to respond to multiple stimuli could enhance the metastatic potential. PMID- 2557333 TI - Substrate phosphorylation catalyzed by the insulin receptor tyrosine kinase. Kinetic correlation to autophosphorylation of specific sites in the beta subunit. AB - The kinetics of insulin-stimulated autophosphorylation of specific tyrosines in the beta subunit of the mouse insulin receptor and activation of receptor kinase catalyzed phosphorylation of a model substrate were compared. The deduced amino acid sequence of the mouse proreceptor was determined to locate tyrosine containing tryptic peptides. Receptor was first incubated with unlabeled ATP to occupy nonrelevant autophosphorylation sites, after which [32P]autophosphorylation at relevant sites and attendant activation of substrate phosphorylation were initiated with [gamma-32P]ATP and insulin. Activation of substrate phosphorylation underwent an initial lag of 10-20 s during which there was substantial 32P-autophosphorylation of tryptic phosphopeptides p2 and p3, but not p1. Following the lag, incorporation of 32P into p1 and the activation of substrate phosphorylation increased abruptly and exhibited identical kinetics. The addition of substrate to the receptor prior to ATP inhibits insulin stimulated autophosphorylation, and consequently substrate phosphorylation. Insulin-stimulated autophosphorylation of the receptor in the presence of substrate inhibited primarily the incorporation of 32P into p1 and drastically inhibited substrate phosphorylation. From Edman radiosequencing of 32P-labeled p1, p2, and p3 and the amino acid sequence of the mouse receptor, the location of each phosphopeptide within the beta subunit was determined. Further characterization of these phosphopeptides revealed that p1 and p2 represent the triply and doubly phosphorylated forms, respectively, of the region within the tyrosine kinase domain containing tyrosines 1148, 1152, and 1153. The doubly phosphorylated forms contain phosphotyrosines either at positions 1148 and 1152/1153 or positions 1152 and 1153. These results indicate that insulin stimulates sequential autophosphorylation of tyrosines 1148, 1152 and 1153, and that the transition from the doubly to the triply phosphorylated forms is primarily responsible for the activation of substrate phosphorylation. PMID- 2557334 TI - Evidence that activation of the respiratory burst oxidase in a cell-free system from human neutrophils is accomplished in part through an alteration of the oxidase-related 67-kDa cytosolic protein. AB - Sodium dodecyl sulfate (SDS) is able to activate the respiratory burst oxidase in a system containing cytosol and solubilized membranes from human neutrophils. When SDS was used to treat cytosol in an otherwise identical system in which the solubilized membrane solution was omitted, the ability of the SDS-treated cytosol to support O2- production was lost in a first-order reaction whose rate constant was virtually identical to the rate constant for the first-order activation of the oxidase in the complete system. Studies with chronic granulomatous disease cytosols showed that the component whose activity was lost was the oxidase related 67-kDa cytosolic protein. The similarity in the rates of oxidase activation and p67 inactivation suggested that the activation of the respiratory burst oxidase in the cell-free system could involve an SDS-mediated alteration in p67. Further support for this idea was provided by kinetic experiments demonstrating that, although the yield of oxidase showed a 2.5-order dependence on cytosol concentration, oxidase activation was nevertheless kinetically irreversible. These two findings, incompatible in general, can be reconciled by a mechanism in which SDS acts specifically on a single oxidase component (i.e. p67), but with an effect that depends on circumstances: oxidase activation, if the SDS-sensitive component is part of a completely assembled oxidase precursor; loss of p67 activity, if not. PMID- 2557335 TI - Purification and characterization of rat liver GTP cyclohydrolase I. Cooperative binding of GTP to the enzyme. AB - GTP cyclohydrolase I, an enzyme that catalyzes the first step in the biosynthetic pathway of tetrahydrobiopterin, has been purified about 38,000-fold to apparent homogeneity from rat liver extract with a yield of 5%. The molecular weight of the enzyme was estimated to be 300,000 by gel filtration on Ultrogel AcA 34. The purified enzyme gave a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis at a position corresponding to a molecular weight of 30,000. N terminal amino acid sequence analysis gave a single amino acid at every step of the Edman degradation up to residue 10. These results suggest that the enzyme is probably a homopolymer. The enzyme showed positive cooperativity with a Hill coefficient of 2.4 at a substrate (GTP) concentration of 10-50 microM. The Vmax value of the enzyme was 45 nmol/min.mg protein. The GTP concentration producing half-maximal velocity was 30 microM at a KCl concentration of 0.1 M. This value increased as the KCl concentration rose, without any change in Vmax or Hill number. Biosynthesis of tetrahydrobiopterin may be controlled by the intracellular level of GTP. PMID- 2557336 TI - Spectroscopic studies of isopenicillin N synthase. A mononuclear nonheme Fe2+ oxidase with metal coordination sites for small molecules and substrate. AB - The nonheme iron oxidase isopenicillin N synthase catalyzes the formation of two new internal bonds in the tripeptide delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D valine (ACV) to form the beta-lactam and thiazolidine rings of isopenicillin N. Concomitantly, O2 is reduced to 2 H2O. The recombinant enzyme from Cephalosporium acremonium (Mr = 38,400), expressed as an apoenzyme in Escherichia coli, binds 1 g atom of Fe2+/mol of enzyme to reconstitute full activity. Mossbauer spectra of the 57Fe-enriched enzyme exhibit parameters (delta = 1.30 mm/s, delta EQ = 2.70 mm/s) which unambiguously show that the active site iron is high spin Fe2+. Anaerobic binding of ACV causes a substantial decrease in the isomer shift parameter delta (delta = 1.10 mm/s, delta EQ = 3.40 mm/s) showing that the substrate perturbs the iron site and makes its coordination environment much more covalent. Nitric oxide (NO) binds to the EPR silent active site iron to give an EPR active species (g = 4.09, 3.95, 2.0; S = 3/2) similar to those of the nitrosyl complexes of many other mononuclear Fe2+-containing enzymes. The rhombicity of the EPR spectrum is increased (g = 4.22, 3.81, 1.99) by anaerobic addition of ACV suggesting that the substrate binds to or near the iron without displacing NO. Interestingly, the enzyme.ACV.NO complex displays an optical spectrum similar to that of ferric rubredoxin in which the iron has only thiol coordination. This suggests that the Fe2+ of the enzyme.ACV.NO complex acquires Fe3+ character and that the cysteinyl thiol moiety of ACV coordinates to the iron. Similar substrate thiol coordination to the iron of the enzyme.ACV complex is the most probable explanation for the large decrease in isomer shift observed. These results provide the first evidence for the direct involvement of iron in this unique O2-dependent reaction and suggest novel roles for iron and oxygen in biological catalysis. PMID- 2557337 TI - Phosphorylation of DARPP-32, a dopamine- and cAMP-regulated phosphoprotein, by casein kinase II. AB - DARPP-32 (dopamine- and cAMP-regulated phosphorprotein, Mr = 32,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis) is an inhibitor of protein phosphatase-1 and is enriched in dopaminoceptive neurons possessing the D1 dopamine receptor. Purified bovine DARPP-32 was phosphorylated in vitro by casein kinase II to a stoichiometry greater than 2 mol of phosphate/mol of protein whereas two structurally and functionally related proteins, protein phosphatase inhibitor-1 and G-substrate, were poor substrates for this enzyme. Sequencing of chymotryptic and thermolytic phosphopeptides from bovine DARPP-32 phosphorylated by casein kinase II suggested that the main phosphorylated residues were Ser45 and Ser102. In the case of rat DARPP-32, the identification of these phosphorylation sites was confirmed by manual Edman degradation. The phosphorylated residues are located NH2-terminal to acidic amino acid residues, a characteristic of casein kinase II phosphorylation sites. Casein kinase II phosphorylated DARPP-32 with an apparent Km value of 3.4 microM and a kcat value of 0.32 s-1. The kcat value for phosphorylation of Ser102 was 5-6 times greater than that for Ser45. Studies employing synthetic peptides encompassing each phosphorylation site confirmed this difference between the kcat values for phosphorylation of the two sites. In slices of rat caudate-putamen prelabeled with [32P]phosphate, DARPP-32 was phosphorylated on seryl residues under basal conditions. Comparison of thermolytic phosphopeptide maps and determination of the phosphorylated residue by manual Edman degradation identified the main phosphorylation site in intact cells as Ser102. In vitro, DARPP-32 phosphorylated by casein kinase II was dephosphorylated by protein phosphatases-1 and -2A. Phosphorylation by casein kinase II did not affect the potency of DARPP-32 as an inhibitor of protein phosphatase-1, which depended only on phosphorylation of Thr34 by cAMP-dependent protein kinase. However, phosphorylation of DARPP-32 by casein kinase II facilitated phosphorylation of Thr34 by cAMP-dependent protein kinase with a 2.2-fold increase in the Vmax and a 1.4-fold increase in the apparent Km. Phosphorylation of DARPP-32 by casein kinase II in intact cells may therefore modulate its phosphorylation in response to increased levels of cAMP. PMID- 2557338 TI - Nuclease protection by Drosophila DNA topoisomerase II. Enzyme/DNA contacts at the strong topoisomerase II cleavage sites. AB - A DNA consensus sequence for topoisomerase II cleavage sites was derived previously based on a statistical analysis of the nucleotide sequences around 16 sites that can be efficiently cleaved by Drosophila topoisomerase II (Sander, M., and Hsieh, T. (1985) Nucleic Acids Res. 13, 1057-1072). A synthetic 21-mer DNA sequence containing this cleavage consensus sequence was cloned into a plasmid vector, and DNA topoisomerase II can cleave this sequence at the position predicted by the cleavage consensus sequence. DNase I footprint analysis showed that topoisomerase II can protect a region of approximately 25 nucleotides in both strands of the duplex DNA, with the cleavage site located near the center of the protected region. Similar correlation between the DNase I footprints and strong topoisomerase II cleavage sites has been observed in the intergenic region of the divergent HSP70 genes. This analysis therefore suggests that the strong DNA cleavage sites of Drosophila topoisomerase II likely correspond to specific DNA-binding sites of this enzyme. Furthermore, the extent of DNA contacts made by this enzyme suggests that eucaryotic topoisomerase II, in contrast to bacterial DNA bacterial DNA gyrase, cannot form a complex with extensive DNA wrapping around the enzyme. The absence of DNA wrapping is probably the mechanistic basis for the lack of DNA supercoiling action for eucaryotic topoisomerase II. PMID- 2557339 TI - Localization of DNA repair synthesis by human cell extracts to a short region at the site of a lesion. AB - Double-stranded bacteriophage M13 DNA molecules were constructed containing a single specifically placed 2-(acetylamino)fluorene adduct or a single 4' hydroxymethyl-4,5',8-trimethylpsoralen monoadduct. These circular DNA molecules were used to analyze in vitro DNA repair synthesis by cell extracts from normal human lymphoid cell lines. Both types of lesions stimulate DNA repair synthesis at the site of the adduct. DNA repair synthesis induced by the 2-(acetyl amino)fluorene adduct took place in the damaged strand and was confined to a region within a 31-base pair restriction fragment around the adduct. PMID- 2557340 TI - Phosphorylation-regulated calmodulin binding to a prominent cellular substrate for protein kinase C. AB - We have evaluated the possibility that a major, abundant cellular substrate for protein kinase C might be a calmodulin-binding protein. We have recently labeled this protein, which migrates on sodium dodecyl sulfate-gel electrophoresis with an apparent Mr of 60,000 from chicken and 80,000-87,000 from bovine cells and tissues, the myristoylated alanine-rich C kinase substrate (MARCKS). The MARCKS proteins from both species could be cross-linked to 125I-calmodulin in a Ca2+ dependent manner. Phosphorylation of either protein by protein kinase C prevented 125I-calmodulin binding and cross-linking, suggesting that the calmodulin-binding domain might be located at or near the sites of protein kinase C phosphorylation. Both bovine and chicken MARCKS proteins contain an identical 25-amino acid domain that contains all 4 of the serine residues phosphorylated by protein kinase C in vitro. In addition, this domain is similar in sequence and structure to previously described calmodulin-binding domains. A synthetic peptide corresponding to this domain inhibited calmodulin binding to the MARCKS protein and also could be cross-linked to 125I-calmodulin in a calcium-dependent manner. In addition, protein kinase C-dependent phosphorylation of the synthetic peptide inhibited its binding and cross-linking to 125I-calmodulin. The peptide bound to fluorescently labeled 5-dimethylaminonaphthalene-1-sulfonyl-calmodulin with a dissociation constant of 2.8 nM, and inhibited the calmodulin-dependent activation of cyclic nucleotide phosphodiesterase with an IC50 of 4.8 nM. Thus, the peptide mimics the calmodulin-binding properties of the MARCKS protein and probably represents its calmodulin-binding domain. Phosphorylation of these abundant, high affinity calmodulin-binding proteins by protein kinase C in intact cells could cause displacement of bound calmodulin, perhaps leading to activation of Ca2+-calmodulin-dependent processes. PMID- 2557341 TI - Transcriptional induction of glucokinase gene by insulin in cultured liver cells and its repression by the glucagon-cAMP system. AB - Primary cultures of rat hepatocytes were used to investigate the regulation of glucokinase gene expression by insulin and glucagon. Insulin added in physiological concentrations to the culture medium causes de novo induction of glucokinase mRNA. The induced plateau is reached 4 to 8 h after insulin addition, and the mRNA level remains high as long as insulin is present. Comparison of the potencies of insulin, proinsulin, and insulin-like growth factor I in this system indicates that induction by insulin is mediated via the insulin receptor. The magnitude of the insulin effect is independent of the extracellular glucose concentration. Run-on transcription assays with isolated nuclei show that the mRNA build up depends primarily on a specific stimulation of glucokinase gene transcription. Glucagon added to hepatocytes together with a supramaximal concentration of insulin prevents induction of glucokinase mRNA in a dose dependent manner. The inhibitory effect of glucagon is mimicked by 8-(4 chlorophenylthio)-cAMP. The effect of this agent has also been tested in hepatocytes first induced for maximal glucokinase gene transcription by culture with insulin alone for 12 h. The transcriptional activity of the gene as measured by run-on assay was completely turned off within 30 min after addition of the cyclic nucleotide. Under these conditions, glucokinase mRNA decays rapidly, with an apparent half-life of 45 min. The mRNA degradation rate was similarly rapid after insulin withdrawal from induced cells. Thus, a cAMP-mediated repression mechanism is a key aspect in the regulation of glucokinase gene transcription in the hepatocyte. Insulin may act by relieving the gene from repression. PMID- 2557342 TI - Generation of the Ca2(+)-independent form of Ca2+/calmodulin-dependent protein kinase II in cerebellar granule cells. AB - Conditions that regulate the generation of the Ca2(+)-independent form of Ca2+/calmodulin-dependent protein kinase II (CaM-kinase II) in cultured rat cerebellar granule cells have been investigated. Under basal conditions, 4-5% of total CaM-kinase II activity, assayed in the presence of Ca2+/CaM, was the Ca2(+) independent form active in the presence of EGTA. Depolarization with 56 mM K+ produced a transient increase to 9% Ca2+ independence within 15 s followed by a decline to 5-6% at 10 min. The divalent cation ionophore ionomycin elicited 10% Ca2+ independence, which remained elevated. Removal of Ca2+ from the Krebs-Ringer medium reduced basal Ca2+ independence to 1-2% and eliminated the elevation in response to K+ depolarization. Inclusion of 5 microM okadaic acid, a protein phosphatase inhibitor, in the incubation medium potentiated the levels of Ca2(+) independent activity of CaM-kinase II. Additional studies in granule cell extracts indicated that there were both okadiac acid-sensitive and -insensitive protein phosphatases involved in the reversal of the Ca2+ independence of CaM kinase II. Phosphopeptide mapping of the CNBr-cleaved 32P-labeled 58-60-kDa subunit of CaM-kinase II revealed that under basal conditions, the kinase contained phosphate in many sites. Conditions that promoted formation of the Ca2(+)-independent form of the kinase increased the 32P incorporation into multiple sites of the kinase. However, there was a good temporal correlation between 32P incorporation into CNBr peptide 1, which contains Thr-287, and generation of the Ca2(+)-independent kinase activity. These results indicate that formation of the Ca2(+)-independent species of CaM-kinase II is dynamically regulated in cerebellar granule cells by Ca2(+)-mobilizing agents and by protein phosphatase activity and is correlated with autophosphorylation of Thr-287. PMID- 2557343 TI - A second type of rat phosphoinositide-specific phospholipase C containing a src related sequence not essential for phosphoinositide-hydrolyzing activity. AB - A fourth type of rat phosphoinositide-specific phospholipase C (PLC IV) has been cloned for cDNA and sequenced. PLC IV is distinct from the other three types of rat PLC (PLC I, II, and III) with respect to primary structure and tissue distribution of its mRNAs. PLC IV contains two homologous regions included commonly in PLC I, II, and III and is most similar to PLC II (identity: 50.2%). PLC IV, in common with PLC II, has a sequence homologous to the N-terminal regulatory domains of nonreceptor tyrosine kinases of the src-family of oncogenes. Using an Escherichia coli expression system, we succeeded in producing active PLC IV in E. coli crude extracts. Various truncation experiments of the PLC IV cDNA revealed that the src-related domain is not necessary for catalytic activity while both domains homologous among PLC I-IV are essential. PLC IV is expressed in various rat tissues and abundant in spleen, suggesting that PLC IV plays a fundamental role in cellular functions such as growth and secretion. PMID- 2557344 TI - Site-directed mutagenesis of a conserved, extracellular aspartic acid residue affects the ouabain sensitivity of sheep Na,K-ATPase. AB - Site-specific mutagenesis was used to study the function of a conserved, extracellular aspartic acid residue from the sheep Na,K-ATPase alpha subunit. This amino acid, Asp-121, is the penultimate residue of the first extracellular domain of the alpha subunit. The border residues of this particular extracellular loop of the alpha subunit have been shown to be determinants of ouabain sensitivity (Price, E. M., and Lingrel, J. B. (1988) Biochemistry 27, 8400-8408). In order to determine if Asp-121 is involved in ouabain binding, five different amino acid substitutions at this position were generated. Four of the five mutant alpha subunits, containing either Asn, Ala, Glu, or Ser in place of Asp-121, conferred ouabain resistance to HeLa cells when expressed in those cells. Cloned sublines of cells selected in ouabain were characterized in terms of ouabain inhibitable cell growth and Na,K-ATPase activity. The cells expressing the mutant Na,K-ATPase alpha subunit containing either Asn, Ala, Glu, or Ser in place of Asp 121 contained a component of Na,K-ATPase activity that was nearly 100-times more resistant to ouabain than the endogenous HeLa (human) or sheep enzyme. Apparently, conservative (Glu for Asp), isosteric (Asn for Asp), and nonconservative (Ala or Ser for Asp) substitutions all significantly decreased ouabain sensitivity. These data suggest that Asp-121 of the sheep Na,K-ATPase alpha subunit participates in the binding interaction between the enzyme and ouabain. PMID- 2557345 TI - Mutations of GS alpha designed to alter the reactivity of the protein with bacterial toxins. Substitutions at ARG187 result in loss of GTPase activity. AB - We have introduced two types of mutations into cDNAs that encode the alpha subunit of Gs, the guanine nucleotide-binding regulatory protein that stimulates adenylyl cyclase. The arginine residue (Arg187) that is the presumed site of ADP ribosylation of Gs alpha by cholera toxin has been changed to Ala, Glu, or Lys. The rate constant for hydrolysis of GTP by all of these mutants is reduced approximately 100-fold compared with the wild-type protein. As predicted from this change, these proteins activate adenylyl cyclase constitutively in the presence of GTP. Despite these substitutions, cholera toxin still catalyzes the incorporation of 0.2-0.3 mol of ADP-ribose/mol of mutant alpha subunit. The sequence near the carboxyl terminus of Gs alpha was altered to resemble those in Gi alpha polypeptides, which are substrates for pertussis toxin. Despite this change, the mutant protein is a poor substrate for pertussis toxin. Although this protein has unaltered rates of GDP dissociation and GTP hydrolysis, its ability to activate adenylyl cyclase in the presence of GTP is enhanced by 3-fold when compared with the wild-type protein but only when these assays are performed after reconstitution of Gs alpha into cyc- (Gs alpha-deficient) S49 cell membranes. PMID- 2557346 TI - Modification of phospholipase C and phospholipase A2 activities during poliovirus infection. AB - The infection of HeLa cells by poliovirus leads to profound alterations in the activities of both phospholipase C and the A23187-stimulated phospholipase A2. As early as the third hour after poliovirus infection, the activity of phospholipase C is enhanced, as measured by the increase in inositol triphosphate (IP3) in the cells. By the fifth hour post-infection there is a 5-fold increase in IP3 in the infected cells. Therefore, the synthesis of the bulk of poliovirus proteins and poliovirus genomes takes place in cells containing a high and sustained increase in IP3. This augmentation in IP3 is dependent on the multiplicity of infection used. Poliovirus gene expression is required to induce the increase in phospholipase C activity, since the presence of cycloheximide or guanidine blocked it. In contrast to the activation of phospholipase C induced by poliovirus, there is a drastic blockade of the A23187-induced phospholipase A2 activity, measured as the release of [3H]arachidonic acid to the medium. This action on phospholipase A2 is dependent on poliovirus gene expression because it was prevented by cycloheximide or 3-methylquercetin. To our knowledge this is the first report analyzing these two activities in animal virus-infected cells. The findings described may help to explain the profound modifications of both membrane permeability and lipid metabolism undergone by poliovirus-infected cells. PMID- 2557348 TI - Naloxone inhibits the in vivo immunosuppressive effects of morphine and thermal injury in mice. AB - The effect of opioids (both exogenous and endogenous) on cell-mediated immune response in normal and thermally injured mice was evaluated with a delayed-type hypersensitivity assay. The administration of morphine sulfate to normal mice resulted in decreased delayed-type hypersensitivity response. This morphine sulfate-induced immunosuppression was prevented by concurrent treatment with the opioid antagonist, naloxone; however, naloxone alone did not alter immune response. Thermally injured mice had a suppressed delayed-type hypersensitivity response that was not further affected by morphine sulfate administration. In contrast, the immunosuppressive effects caused by burn injury, alone or in combination with the administration of morphine sulfate, were not observed in the presence of naloxone as measured by delayed-type hypersensitivity response. These results suggest that opioids depress cellular immune response and may play a role in immune dysfunction that follows thermal injury. PMID- 2557347 TI - The human 5-HT1A receptor expressed in HeLa cells stimulates sodium-dependent phosphate uptake via protein kinase C. AB - Regulation of phosphate uptake was studied in HeLa cell lines after transfection with DNA encoding the human 5-HT1A receptor. Phosphate uptake was saturable and greater than 90% sodium-dependent, with Vmax approximately 30-35% without changing Km. Treatment with 5-HT or the 5-HT1A-specific agonist 8-OH-2-(di-n propylamino)1,2,3,4-tetrahydronaphthalene increased Vmax approximately 40% without affecting Km. This effect was blocked by pretreatment with the 5-HT1 antagonists, methiothepine and spiperone, or pertussis toxin. Surprisingly, the stimulation was not secondary to an inhibition of adenylyl cyclase because 5-HT stimulated phosphate uptake approximately 20% in the presence of 1 mM 8-Br-cAMP. Rather, the primary pathway linked to the stimulation of phosphate uptake involved activation of protein kinase C because (i) 5-HT measurably activated protein kinase C in these cells, (ii) activators of protein kinase C (phorbol esters and diacylglycerol analogues) stimulated phosphate uptake in these cells (iii) the half-maximal doses for 5-HT-induced phosphatidylinositol hydrolysis and stimulation of phosphate uptake were virtually equivalent, and both effects were equally sensitive to pertussis toxin, and (iv) the stimulation was markedly attenuated in cells made deficient in protein kinase C. These results demonstrate that the stimulation of phosphatidylinositol hydrolysis by the 5-HT1A receptor can generate physiologically measurable effects on cellular transport and suggest that such accessory pathways may play a prominent role in signal transduction. PMID- 2557349 TI - 4-aminopyridine restores conduction in heat-blocked sciatic branches. AB - We induced partial conduction block in rat sural and posterior tibial nerves by heating a short (5 mm) segment of the midportions of these branches to 45 degrees C until the amplitude of the elicited compound action potential was reduced by at least 50%, an effect that appeared to be irreversible without specific treatment. Excised branches studied in vitro were compared with branches heated in situ with an intact blood supply. The potassium channel blocker 4-aminopyridine (4-AP, 5 mM) was applied topically to the heated segments at intervals after injury. The amplitude of the compound action potential was restored to the control level in all preparations treated within 10 minutes after the induction of conduction block. The reversal appeared to be more rapid in the in vivo preparations, and it persisted for at least 60 minutes. The data suggest that paranodal potassium channel exposure may accompany direct thermal injury to the peripheral nerve. PMID- 2557350 TI - Saccharomyces cerevisiae and Schizosaccharomyces pombe contain a homologue to the 54-kD subunit of the signal recognition particle that in S. cerevisiae is essential for growth. AB - We have isolated and sequenced genes from Saccharomyces cerevisiae (SRP54SC) and Schizosaccharomyces pombe (SRP54sp) encoding proteins homologous to both the 54 kD protein subunit (SRP54mam) of the mammalian signal recognition particle (SRP) and the product of a gene of unknown function in Escherichia coli, ffh (Romisch, K., J. Webb, J. Herz, S. Prehn, R. Frank, M. Vingron, and B. Dobberstein. 1989. Nature (Lond.). 340:478-482; Bernstein H. D., M. A. Poritz, K. Strub, P. J. Hoben, S. Brenner, P. Walter. 1989. Nature (Lond.). 340:482-486). To accomplish this we took advantage of short stretches of conserved sequence between ffh and SRP54mam and used the polymerase chain reaction (PCR) to amplify fragments of the homologous yeast genes. The DNA sequences predict proteins for SRP54sc and SRP54sp that are 47% and 52% identical to SRP54mam, respectively. Like SRP54mam and ffh, both predicted yeast proteins contain a GTP binding consensus sequence in their NH2-terminal half (G-domain), and methionine-rich sequences in their COOH-terminal half (M-domain). In contrast to SRP54mam and ffh the yeast proteins contain additional Met-rich sequences inserted at the COOH-terminal portion of the M-domain. SRP54sp contains a 480-nucleotide intron located 78 nucleotides from the 5' end of the open reading frame. Although the function of the yeast homologues is unknown, gene disruption experiments in S. cerevisiae show that the gene is essential for growth. The identification of SRP54sc and SRP54sp provides the first evidence for SRP related proteins in yeast. PMID- 2557351 TI - Meeting of the apical and basolateral endocytic pathways of the Madin-Darby canine kidney cell in late endosomes. AB - Electron microscopic approaches have been used to study the endocytic pathways from the apical and basolateral surface domains of the polarized epithelial cell, MDCK strain I, grown on polycarbonate filters. The cells were incubated at 37 degrees C in the presence of two distinguishable markers administered separately to the apical or the basolateral domain. Initially each marker was visualized within distinct apical or basolateral peripheral endosomes. However, after 15 min at 37 degrees C, both markers were observed within common perinuclear structures. The compartment in which meeting first occurred was shown to be a late endosome (prelysosome) that labeled extensively with antibodies against the cation independent mannose-6-phosphate receptor (MPR) on cryosections. With increasing incubation times, markers passed from these MPR-positive structures into a common set of MPR-negative lysosomes that were mainly located in the apical half of the cell. A detailed quantitative analysis of the endocytic pathways was carried out using stereological techniques in conjunction with horseradish peroxidase and acid phosphatase cytochemistry. This enabled us to estimate the absolute volumes and membrane surface areas of the endocytic organelles involved in apical and basolateral endocytosis. PMID- 2557352 TI - Intracellular maturation and transport of the SV5 type II glycoprotein hemagglutinin-neuraminidase: specific and transient association with GRP78-BiP in the endoplasmic reticulum and extensive internalization from the cell surface. AB - The hemagglutinin-neuraminidase (HN) glycoprotein of the paramyxovirus SV5 is a type II integral membrane protein that is expressed at the infected cell surface. The intracellular assembly and transport of HN in CV1 cells was examined using conformation-specific HN mAbs and sucrose density sedimentation analysis. HN was found to oligomerize with a t1/2 of 25-30 min and these data suggest the oligomer is a tetramer consisting primarily of two noncovalently associated disulfide linked dimers. As HN oligomers could be found that were sensitive to endoglycosidase H digestion and oligomers formed in the presence of the ER to the Golgi complex transport inhibitor, carbonylcyanide m-chlorophenylhydrazone (CCCP), these data are consistent with HN oligomerization occurring in the ER. Unfolded or immature HN molecules that could not be recognized by conformation specific antibodies were found to specifically associate with the resident ER protein GRP78-BiP. Immunoprecipitation of BiP-HN complexes with an immunoglobulin heavy-chain binding protein (BiP) antibody indicated that newly synthesized HN associated and dissociated from GRP78-BiP (t1/2 20-25 min) in an inverse correlation with the gain in reactivity with a HN conformation-specific antibody, suggesting that the transient association of GRP78-BiP with immature HN is part of the normal HN maturation pathway. After pulse-labeling of HN in infected cells, it was found that HN is rapidly turned over in cells (t1/2 2-2.5 h). This led to the finding that the vast majority of HN expressed at the cell surface, rather than being incorporated into budding virions, is internalized and degraded after localization to endocytic vesicles and lysosomes. PMID- 2557353 TI - Translational diffusion of class II major histocompatibility complex molecules is constrained by their cytoplasmic domains. AB - Site-directed mutagenesis in vitro was used to introduce stop codons in the genomic DNA of the alpha and beta chains of the murine class II major histocompatibility complex antigen, I-Ak. Mutated DNA was transfected into B lymphoma cells that were then selected by neomycin resistance and for their ability to express I-Ak molecules on their plasma membrane. The translational diffusion coefficient (Dlat) of I-Ak molecules composed of a wild-type beta chain paired with an alpha chain missing either 6 or 12 amino acids from the cytoplasmic domain is on the average threefold higher than the Dlat of wild-type I-Ak molecules as measured by fluorescence photobleaching and recovery. The removal of 12 amino acids from the cytoplasmic domain of the beta chain did not change the Dlat value from that of wild-type I-Ak if the truncated beta chain was paired with a wild-type alpha chain. Removing all amino acids of the cytoplasmic domains of both the alpha and beta chains resulted in a 10-fold increase in the Dlat, the highest value for any of the truncated I-Ak molecules tested. These data indicate that the carboxy-terminal six amino acids of the cytoplasmic domain of the alpha chain and the six plasma membrane-proximal amino acids of the beta chain are important in constraining the translational diffusion of I-Ak molecules in the plasma membrane. PMID- 2557354 TI - Sequence and tissue distribution of a second protein of hepatic gap junctions, Cx26, as deduced from its cDNA. AB - While a number of different gap junction proteins have now been identified, hepatic gap junctions are unique in being the first demonstrated case where two homologous, but distinct, proteins (28,000 and 21,000 Mr) are found within a single gap junctional plaque (Nicholson, B. J., R. Dermietzel, D. Teplow, O. Traub, K. Willecke, and J.-P. Revel. 1987. Nature [Lond.]. 329:732-734). The cDNA for the major 28,000-Mr component has been cloned (Paul, D. L. 1986. J. Cell Biol. 103:123-134) (Kumar, N. M., and N. B. Gilula. 1986. J. Cell Biol. 103:767 776) and, based on its deduced formula weight of 32,007, has been designated connexin 32 (or Cx32 as used here). We now report the selection and characterization of clones for the second 21,000-Mr protein using an oligonucleotide derived from the amino-terminal protein sequence. Together the cDNAs represent 2.4 kb of the single 2.5-kb message detected in Northern blots. An open reading frame of 678 bp coding for a protein with a calculated molecular mass of 26,453 D was identified. Overall sequence homology with Cx32 and Cx43 (64 and 51% amino acid identities, respectively) and a similar predicted tertiary structure confirm that this protein forms part of the connexin family and is consequently referred to as Cx26. Consistent with observations on Cx43 (Beyer, E. C., D. L. Paul, and D. A. Goodenough. 1987. J. Cell Biol. 105:2621-2629) the most marked divergence between Cx26 and other members of the family lies in the sequence of the cytoplasmic domains. The Cx26 gene is present as a single copy per haploid genome in rat and, based on Southern blots, appears to contain at least one intron outside the open reading frame. Northern blots indicate that Cx32 and Cx26 are typically coexpressed, messages for both having been identified in liver, kidney, intestine, lung, spleen, stomach, testes, and brain, but not heart and adult skeletal muscle. This raises the interesting prospect of having differential modes of regulating intercellular channels within a given tissue and, at least in the case of liver, a given cell. PMID- 2557356 TI - Transforming growth factors-beta 1 and beta 2 are mitogens for rat Schwann cells. AB - Transforming growth factor-beta 1 (TGF-beta 1) and TGF-beta 2 were found to be potent mitogens for purified rat Schwann cells, each stimulating DNA synthesis in quiescent cells and also increasing their proliferation rate. Half-maximal stimulation of DNA synthesis occurred at approximately 0.1 ng/ml TGF-beta 1 or TGF-beta 2. Mitogenic stimulation by TGF-beta 1 and TGF-beta 2 was enhanced by forskolin, which activates adenylate cyclase, at concentrations up to 0.5 microM forskolin. However, at 5 microM forskolin, the synergistic interaction between forskolin and TGF-beta 1 was abolished. These results are in contrast to the observed synergy between forskolin and another Schwann cell mitogen, glial growth factor (GGF). Both 0.5 and 5 microM forskolin were found to enhance the stimulation of DNA synthesis by partially purified GGF (GGF-CM). As well as being functionally distinct, TGF-beta 1 and GGF-CM activities were also physically separable by chromatography on a Superose 12 gel permeation column. Thus, TGF beta 1 and beta 2 are rat Schwann cell mitogens, and Schwann cells are one of the few normal cell populations to respond mitogenically to TGF-beta. PMID- 2557357 TI - MSH inhibits growth in a line of amelanotic hamster melanoma cells and induces increases in cyclic AMP levels and tyrosinase activity without inducing melanogenesis. AB - In Bomirski Ab amelanotic hamster melanoma cells, L-tyrosine and/or L-dopa induce increases in tyrosinase activity as well as synthesis of melanosomes and melanin. L-tyrosine also modifies melanocyte-stimulating hormone (MSH) binding. In this paper we show that in the Bomirski amelanotic melanoma system MSH and agents that raise intracellular cyclic AMP induce dendrite formation, inhibit cell growth, and cause substantial increases in tyrosinase activity without inducing melanin synthesis. Tyrosinase activity is detected only in broken cell preparations, or cytochemically in fixed cells. In the continued absence of mature melanosomes, the induced enzyme remains in elements of the trans-Golgi reticulum. Comparative measurements of cyclic AMP in amelanotic and tyrosine-induced melanotic cells show similar basal levels. L-tyrosine and L-dopa have little or no effect, whereas MSH may cause a 1000% peak increase in cyclic AMP levels both in amelanotic and melanotic cells. None of these agents influences cyclic GMP or inositol trisphosphate (InsP3) levels. In agreement with the InsP3 assays, phorbol ester (TPA) has no effect on melanization, tyrosinase activity or cell proliferation. In conclusion, in the Bomirski amelanotic melanoma, MSH induces only partial cell differentiation associated with raised levels of cyclic AMP. Induction of melanosome synthesis and melanization by L-tyrosine or L-dopa appear to follow pathways unrelated to cyclic AMP, cyclic GMP or InsP3. PMID- 2557358 TI - Lectin binding of F9 embryonal carcinoma cells: evidence for population heterogeneity and developmentally regulated high-Mr cell surface proteins. AB - Undifferentiated F9 embryonal carcinoma (EC) cells bound fluorochrome-coupled Helix pomatia agglutinins (HPA) and peanut agglutinins (PNA) homogeneously, but were distinctly heterogeneous in their binding of Dolichos biflorus agglutinin (DBA) conjugates. Upon chemically induced differentiation the proportion of cells binding the DBA conjugates increased, but a distinct heterogeneity in the intensity of binding remained among the parietal endoderm (PE)-like F9 derivatives. These cells were heterogeneous in their binding of HPA conjugates as well, and many of them failed to bind PNA conjugates, apparently due to sialylation of the PNA-binding sites. Electrophoretic analysis of lectin-binding glycoproteins in the detergent-soluble fraction of the cells revealed the appearance of a doublet of polypeptides of Mr 300,000-400,000 upon differentiation induced by retinoic acid (RA). In addition, an Mr 220,000 polypeptide appeared upon differentiation induced by RA and dibutyryl cyclic AMP (dbcAMP). These polypeptides were obtained from both metabolically labelled and surface-labelled cells. A major secreted glycoprotein, which comigrated with laminin, bound to DBA. This suggests that laminin secreted by the differentiated F9 derivatives contains O-glycosidic saccharides. The results show that even though differentiation of F9 cells leads to changes in their binding of fluorochrome-coupled lectins, these lectin conjugates reveal distinct population heterogeneity among undifferentiated and differentiated F9 cells and are hence likely to be of limited value in the characterization of individual cells. At the whole cell population level, on the other hand, affinity binding to lectins reveals the appearance of high-Mr cell surface proteins in differentiating F9 cells. PMID- 2557359 TI - Intracellular transport of the glycoprotein of VSV is inhibited by CCCP at a late stage of post-translational processing. AB - The appearance of newly synthesized glycoprotein (G) of vesicular stomatitis virus at the surface of infected BHK cells is inhibited reversibly by treatment with carbonylcyanide m-chlorophenylhydrazone (CCCP). Under the conditions used, CCCP treatment depleted the cellular ATP levels by 40-60%, consistent with inhibition of transport at energy-requiring stages. The G protein that accumulates in cells treated with CCCP is heterogeneous. Most of it is larger than the newly synthesized G protein, is acylated with palmitic acid, and is resistant to endoglycosidase H (Endo H). Most of the arrested G protein is also sensitive to digestion with neuraminidase, indicating that it has undergone at least partial sialylation. A minority of G protein accumulates under these conditions in a less-mature form, suggesting its inability to reach the mid-Golgi compartment. The oligosaccharides of this G protein are Endo-H-sensitive and seem to be partly trimmed. Whereas sialylated G protein was arrested intracellularly, fucose-labelled G protein was able to complete its transport to the cell surface, indicating that a late CCCP-sensitive step separates sialylation from fucosylation. These post-translational modifications indicate that G protein can be transported as far as the trans-Golgi in the presence of CCCP and is not merely arrested in the endoplasmic reticulum. PMID- 2557355 TI - PDGF and intracellular signaling in the timing of oligodendrocyte differentiation. AB - In the rat optic nerve, bipotential O-2A progenitor cells give rise to oligodendrocytes and type 2 astrocytes on a precise schedule. Previous studies suggest that PDGF plays an important part in timing oligodendrocyte development by stimulating O-2A progenitor cells to proliferate until they become mitotically unresponsive to PDGF, stop dividing, and differentiate automatically into oligodendrocytes. Since the loss of mitotic responsiveness to PDGF has been shown not to be due to a loss of PDGF receptors, we have now examined the possibility that the unresponsiveness results from an uncoupling of these receptors from early intracellular signaling pathways. We show that (a) although PDGF does not stimulate newly formed oligodendrocytes to synthesize DNA, it induces an increase in cytosolic Ca2+ in these cells; (b) a combination of a Ca2+ ionophore plus a phorbol ester mimics the effect of PDGF, both in stimulating O-2A progenitor cell division and in reconstituting the normal timing of oligodendrocyte differentiation in culture; and (c) the same combination of drugs does not stimulate newly formed oligodendrocytes to proliferate, even in the presence of PDGF or dibutyryl cAMP. The most parsimonious explanation for these results is that O-2A progenitor cells become mitotically unresponsive to PDGF because the intracellular signaling pathways from the PDGF receptor to the nucleus are blocked downstream from the receptor and some of the early events that are triggered by receptor activation. PMID- 2557360 TI - The glycoprotein of VSV accumulates in a distal Golgi compartment in the presence of CCCP. AB - The post-translational modifications of the G protein of vesicular stomatitis virus, described in the preceding paper, indicate that its transport is arrested by carbonylcyanide m-chlorophenylhydrazone (CCCP) in or near the trans-Golgi. Immunofluorescence microscopy of BHK-21 cells infected with vesicular stomatitis virus and treated with CCCP shows an accumulation of G protein in the Golgi area. In the same cells, the morphology of wheat germ agglutinin (WGA)-staining structures in the perinuclear region is aberrant. Using anti-BiP antibody, there is no obvious change in the structure of the endoplasmic reticulum. Electron microscopy reveals that the aberrant structures in the perinuclear region result from dilation of Golgi cisternae and accumulation of large vacuoles near the Golgi stack. The appearance of these aberrant structures is dose-dependent and they disappear after the protonophore is removed. The vast majority of the vacuoles accumulate on the trans side of the Golgi stack. A small fraction of them contain the marker enzyme thiamine pyrophosphatase (TPPase). By immunoelectron microscopy, most of the vacuoles contain G protein. We conclude that most of the Golgi-associated vacuoles are derived from a distal Golgi transport compartment, possibly the trans-Golgi reticulum, and that CCCP reversibly inhibits the transport of newly synthesized G protein through this distal compartment. PMID- 2557361 TI - Interaction of mouse sperm with purified sperm receptors covalently linked to silica beads. AB - We describe a solid-phase assay that has permitted further analysis of zona pellucida glycoprotein, ZP3, as sperm receptor and acrosome reaction-inducer during fertilization in mice. The assay employs silica beads that contain epoxy groups to which purified, mouse oocyte ZP3 is covalently linked (ZP3-beads). ZP3 beads were characterized, e.g. by whole-mount autoradiography and flow cytofluorometry, incubated with capacitated mouse sperm under a variety of conditions, and the extent of sperm binding determined by light microscopy. Results of experiments presented suggest the following: (1) sperm bind specifically to ZP3-beads, but not to silica beads either exposed to 2 aminoethanol or derivatized with oocyte ZP2, fetuin or bovine serum albumin. (2) In nearly all cases, only one sperm binds per ZP3-bead and binding occurs via the sperm head. (3) The extent of sperm binding to ZP3-beads is dependent on ZP3 and sperm concentrations, as well as on incubation time and temperature. (4) Sperm binding to ZP3-beads is unaffected by antibodies directed against ZP3, but is inhibited in a reversible manner by treatment of ZP3-beads with galactose oxidase. (5) Only acrosome-intact sperm bind to ZP3-beads but, once bound, sperm can undergo the acrosome reaction, which results in their release from ZP3-beads. (6) Islet-activating protein and 3-quinuclidinyl benzilate, two inhibitors of the zona pellucida-induced acrosome reaction, prevent sperm bound to ZP3-beads from undergoing the acrosome reaction. These results confirm and extend previous studies of sperm-egg interaction in mice, and suggest that the solid-phase assay will be useful for both cellular and biochemical analyses of mammalian fertilization. PMID- 2557362 TI - Analytical high-performance liquid chromatography system for separation of components in nonoxynol-9 spermicidal agents. PMID- 2557363 TI - Ammonia-treated alfalfa silage for lactating dairy cows. AB - Alfalfa was treated (3.3% of forage wet weight) with an aqua-NH3 (14% N) mix prior to ensiling in 2 consecutive yr. In the 1st yr, NH3-treated silage containing 30.5% DM was higher in pH, N, NH3N, acetic acid and butyric acid contents, and lower in amino acids, soluble carbohydrates, and lactic acid than two untreated silages having 32.8 and 47.5% DM. Milk composition and DM intake did not differ among treatments when these silages constituted 50% of the dietary DM and were fed to lactating cows. In situ and in vitro, but not in vivo, digestion of NDF was greater for NH3 silage. Dry matter content of NH3-treated silage in the 2nd yr was 36.1% and that of the two untreated silages was 37.8 and 54.9%. Silage N, NH3N, and acetic acid content were greater in treated silage. However, pH and butyric acid content were similar among treatments and lactic acid was greater for treated silage. Similar to findings in the 1st yr, milk production and DM intake were not different among treatments. Adding NH3 to alfalfa decreased proteolysis; however, when DM was low (30.5%), the buffering effect of NH3 apparently resulted in increased clostridial fermentation. This did not occur when silage DM was increased to 36.1%, presumably due to higher osmotic pressure. Ammonia addition enhanced fiber digestibility but had no effect on milk production. PMID- 2557364 TI - Matching alcoholics to coping skills or interactional therapies: posttreatment results. AB - This study tested the hypothesis that patients could be matched to effective treatments on the basis of certain pretreatment characteristics. Specifically, it was hypothesized that those Ss who showed more sociopathy, more psychopathology, and greater neuropsychological impairment would have better outcomes when treated with coping skills training and, conversely, that those with less impairment in these areas would have better outcomes with interactional treatment. Ninety-six male and female Ss were recruited from an inpatient alcoholism treatment program and randomly assigned to 1 of these 2 types of aftercare group treatment. Linear and logistic regression analyses partially confirmed the hypotheses. Coping skills training was more effective for Ss higher in sociopathy or psychopathology, and interactional therapy was more effective for Ss lower in sociopathy. Generally, both treatments appeared equally effective for Ss lower in psychopathology. Contrary to expectations, those more neuropsychologically impaired appeared to have better outcomes after interactional therapy. PMID- 2557365 TI - Superoxide anion in polymorphonuclear leukocytes of hairless rats suffering from magnesium-deficiency dermatitis. AB - Clinical and histological studies were made on magnesium-deficiency dermatitis produced on a new strain of hairless rats. Superoxide anion generation in polymorphonuclear leukocytes (PMNs) and the inhibitory effect of some antioxidants and anti-inflammatory agents against the elicited dermatitis were also examined. From the detection of increased superoxide production and effective actions of inhibitors including superoxide dismutase, cepharanthine, 4 4'-diamono-diphenylsulphone and 5,4,11,14-eicosatetraynoic acid (ETYA), it is indicated that superoxide anion or related activated oxygen may be responsible for magnesium-deficiency dermatitis as an inflammatory mediator and that the dermatitis may be related to lipoxygenase metabolites derived from arachidonic acid. PMID- 2557366 TI - Palmitate-derivatized antibodies can function as surrogate receptors for mediating specific cell-cell interactions. AB - The observation that exogenously supplied agents can bypass intrinsic recognition mechanisms and facilitate cellular conjugation has led to valuable insights into the mechanisms of cell function. A common feature of currently available cellular conjugation agents is their reliance on endogenous membrane molecules on both cell types as anchors for cellular interactions. In this report, we describe a method for incorporating palmitate-derivatized antibody molecules onto cell membranes where they function as 'surrogate receptors' (SR) for mediating specific cellular interactions. In this system, SR are attached to the plasma membrane by insertion of the palmitate hydrocarbon chains into the outer leaflet of the phospholipid bilayer. Therefore, the palmitate anchor bypasses the requirement for FcR or other endogenous membrane proteins in antibody-dependent cellular conjugation. Due to this mode of attachment, which is similar to that of phosphatidylinositol (PI) anchored proteins, SR-mediated cellular interactions are likely to be reminiscent of native receptor-induced conjugation, enabling SR to cooperate with endogenous target recognition structures in receptor-ligand interactions at the cell-cell interface. PMID- 2557367 TI - Influence of standard and novel LTB4 analogs on human neutrophil chemotaxis measured by the multiwell cap assay. AB - Standard and novel LTB4 analogs were tested for neutrophil chemoattractant activity using the multiwell cap assay (Evans et al. (1986) Biosc. Rep. 6, 1041). The assay uses disposable equipment and measures chemotaxis by the number of cells able to migrate across the full thickness of cellulose nitrate filters. Under standard conditions (90 min incubation at 37 degrees C in buffer containing 2% bovine albumin), LTB4 and 6-cis-LTB1 had EC50 values of 3.5 and 15,000 nM, respectively. 20-hydroxy-LTB4 was equipotent with LTB4 and exhibited a similar biphasic chemotactic response, however, only one third of the number of cells migrated through the filter. 20-carboxy-LTB4 was inactive up to 1,000 nM. 5 desoxy-((6,7)-cis-cyclopropyl)-LTB2, (6,7)-benzo-LTB2 and 5-desoxy-(8,10)-LTB2 had EC50 values of 11,300, 50,000 and 84,000 nM, respectively. Checkerboard analysis indicated a chemokinetic component of 42% for LTB4 at a concentration causing peak chemotaxis. Reduction of albumin in the buffer to 0.5% increased the apparent potencies of LTB4 and 6-cis-LTB1 five-fold. Since LTB4 is a mediator of inflammation, various anti-inflammatory agents were tested at peak concentrations observed in vivo for in vitro inhibition of LTB4-stimulated chemotaxis in the presence of 0.5% albumin. Under the conditions of the assay, chloroquine diphosphate, dexamethasone, indomethacin, penicillamine, piroxicam and diclofenac sodium were inactive; gold sodium thiomalate was inhibitory (IC50 = 20 microM). PMID- 2557368 TI - Oxygen-independent antimicrobial action in sphingosine-treated neutrophils. AB - Sphingosine is reported to inhibit the oxidative burst and superoxide anion production of human polymorphonuclear neutrophils (PMN) phagocytosing in atmospheric oxygen (Wilson et al., 1986). We have confirmed its effect on superoxide production and examined the antimicrobial phagocytic capacity of PMN treated with sphingosine, comparing them with PMN, untreated but phagocytosing either under anaerobic conditions or in atmospheric oxygen. Sphingosine just like anaerobiosis partially inhibited, but did not eliminate, the bactericidal activity of PMN when compared to non-treated aerobic cells. In fact, sphingosine treated PMN mimicked killing of Staphylococcus aureus (S. aureus) and Serratia marcescens (S. marcescens) due to anaerobic PMN. Moreover, our results with Salmonella typhimurium and sphingosine-treated cells duplicated results this laboratory published previously about comparative killing of Salmonella in aerobic versus anaerobic neutrophils. In these studies sphingosine-treated PMN took up bacteria as avidly as untreated PMN and retained their viability, as assessed by trypan blue exclusion. While sphingosine should not be completely substituted for anaerobic studies, it is a convenient screening reagent for the study of non-oxidative killing mechanisms of PMN. Results achieved with anaerobic and with sphingosine-treated cells suggest that O2-independent antimicrobial action is substantially more powerful than has been generally acknowledged. PMID- 2557369 TI - Enterovirus-specific IgM in the diagnosis of meningitis. AB - Samples of serum from 557 patients with a clinical diagnosis of meningitis or encephalitis and referred to the Epsom Public Health Laboratory during a period of 3 years were tested for enterovirus-specific IgM in a mu capture enzyme-linked immunosorbent assay (ELISA). Enterovirus-specific IgM was detected in 45% samples from all age groups. In the 3-5-year age group, 67% specimens were positive. A notable male predominance (73%) was seen in the age group 0-15 years. As predicted, a seasonal increase in incidence was found in the summer and autumn months. Data from a questionnaire sent to the referring laboratories showed only a 5% enterovirus isolation rate from cerebrospinal fluids when isolation of a virus was attempted. The enterovirus IgM ELISA is a sensitive economical and rapid method for use in the diagnosis of viral meningitis. PMID- 2557370 TI - Ganciclovir infusion at home. PMID- 2557371 TI - Direct ESR measurement of free radicals in mouse pancreatic lesions. AB - In this experiment, free radicals in the pancreas of endotoxemia and ethionine induced acute pancreatitis in mice were attempted to be detected directly by ESR spectroscopy, using 77 K freeze-trapping and 25 degrees C DMPO spin trapping techniques. In the 77 K freeze-trapping method, Mn (II) ion and R-00. radical were detected in endotoxemia and ethionine induced pancreatic lesions. The heme NO radical was observed at 6 and 24 h after isolation of the normal pancreas, and signal intensity was increased with time. This finding supports that ESR spectroscopy is a useful method for detecting the tissue degeneration process from ischemia to necrosis. Using the DMPO spin trapping technique (25 degrees C), 6-line was detected at 6 h after intraperitoneal administration of E. coli in the model of endotoxemia, and 3- and 6-lines and a signal suggestive of DMPO-OH adduct were noted at 12 and 24 h in ethionine pancreatitis. These findings suggest that impaired pancreatic tissues exist in a considerably oxidative environment and oxygen derived free radicals may be considered to play an important role in the development of pancreatic lesions. PMID- 2557372 TI - [A case of the hepatoma associated with hypergastrinemia]. PMID- 2557374 TI - Spontaneous maturation of follicular oocytes in Rana dybowskii in vitro: seasonal influences, progesterone production and involvement of cAMP. AB - Seasonal and hormonal influences regulating oocyte maturation (germinal vesicle breakdown, GVBD) in ovarian follicles of Rana dybowskii were investigated. During the early winter (Dec.-Jan.) GVBD occurred at a low incidence following in vitro culture of intact follicles. Addition of progesterone of frog pituitary homogenate (FPH) to such follicles induced oocyte maturation, whereas IBMX or forskolin inhibited hormone-induced oocyte maturation. The time course of spontaneous in vitro maturation varied markedly with the seasons and between animals. Follicles isolated from the ovaries in early February required 21-24 hours of culture to mature spontaneously, and addition of FPH or progesterone to the culture medium markedly accelerated the time course of GVBD. In contrast, follicles isolated in late February matured very rapidly (less than 6 hours), and FPH or progesterone were ineffective in accelerating the time course of GVBD. IBMX and forskolin separately or in combination stimulated follicular progesterone production, which resembled that seen following FPH stimulation. FPH addition to such follicles shifted the steroid peak to the left (accelerated) and increased the absolute amount of hormone detected in late-maturing follicles (50% GVBD, about 18 hours) but not in rapidly maturing follicles (50% GVBD, 3 hours). In contrast to other amphibians, a high incidence of spontaneous oocyte maturation occurred during in vitro culture. Essentially all animals exhibited spontaneous maturation during the normal breeding season, even those animals collected in the early winter and kept in artificial hibernation at 4 degrees C for extended periods. PMID- 2557373 TI - Characterization of apoA-I-containing lipoprotein subpopulations secreted by HepG2 cells. AB - Recent immunoaffinity studies demonstrate two populations of high density lipoprotein (HDL) particles: one contains both apolipoprotein (apo) A-I and A-II [Lp(A-I w A-II)], and the other contains apoA-I but no A-II [Lp(A-I w/o A-II)]. To investigate whether these two populations are derived from different precursors, we applied sequential immunoaffinity chromatography to study the lipoprotein complexes in HepG2 conditioned serum-free medium. The apparent secretion rates of apoA-I, A-II, E, D, A-IV, and lecithin:cholesterol acyltransferase (LCAT) were 4013 +/- 1368, 851 +/- 217, 414 +/- 64, 171 +/- 51, 32 +/- 14, and 2.9 +/- 0.7 ng/mg cell protein per 24 h, respectively (n = 3-5). Anti-A-II removed all apoA-II but only 39 +/- 5% (n = 5) apoA-I from the medium. These HepG2 Lp(A-I w A-II) also contained 31 +/- 1% (n = 5) of the apoD and 82 +/ 2% (n = 3) of the apoE in the medium. The apoE existed both as E and E-A-II complex. Lipoproteins isolated from the apoA-II-free medium by anti-A-I contained, besides apoA-I, 60 +/- 3% of the medium apoD and trace quantities of apoE. The majority of HepG2 apoA-IV (78 +/- 4%) (n = 3) and LCAT (85 +/- 6%) (n = 3) was not associated with either apoA-I or A-II. HepG2 Lp(A-I w A-II) contained relatively more lipids than Lp(A-I w/o A-II) (45 vs. 37%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557375 TI - Chemotherapy combined with surgery in successful treatment of hepatoblastoma. AB - Four patients presented with large intra-abdominal masses. Three patients were diagnosed as having hepatoblastomas of the right hepatic lobe, one of the left lobe. Initially these tumors were unresectable, and after biopsy each patient was treated with a standardized chemotherapy protocol. All tumors decreased remarkably in size and were resected successfully. Postoperative chemotherapy was then instituted and, as of this date, three of the children appear to be disease free; the fourth had a pulmonary metastatic lesion completely resected. Contrary to the dismal past history of patients with initially unresectable hepatoblastomas, treatment with preoperative chemotherapy, surgery and postoperative chemotherapy has had most rewarding results. PMID- 2557376 TI - AIDS: the HIV epidemic. A time of transition. AB - The surveillance of clinical cases of acquired immune deficiency syndrome (AIDS) in the United States began in June 1981 when the first case was reported. Since then, state and federal public health officials have continuously monitored progression of the epidemic by the number of persons reported with diagnosed clinical AIDS. The human immunodeficiency virus (HIV) that causes AIDS was identified in 1983 and an antibody test was licensed by the Food and Drug Administration in 1985. Recent studies have shown that the time from HIV infection to development of clinical AIDS is an average of seven or more years. Consequently, the reported cases of clinical AIDS reflect the severity of the epidemic an average of seven years ago or more, not now, and certainly not in the future. The AIDS epidemic is in reality an HIV epidemic. The number of persons with HIV infections is a better measure of the present status and future course of the disease. This paper discusses the development of HIV prevalence and incidence studies and illustrates the use of these data to predict the future number of persons with clinical AIDS and the economic impact of the epidemic. PMID- 2557378 TI - Cloning in Escherichia coli of the enterotoxin gene from Clostridium perfringens type A. AB - A 26 bp DNA probe has been constructed with minimal degeneracy to the protein sequence for Clostridium perfringens enterotoxin. The probe has been hybridized against a 6-10 kb chromosomal bank from C. perfringens 8239, prepared as a HindIII partial digest in pHG165. From this survey a clone has been identified containing a 6.8 kb DNA insert with strong hybridization to the probe. Direct plasmid sequencing has identified a translational reading frame within this clone which correlates with the known protein sequence for the type A enterotoxin. DNA sequences 5' to this open reading frame and containing the putative transcriptional control regions show areas of significant homology with regions upstream from the ATG codon of the tetanus toxin gene. PMID- 2557377 TI - Anaerobic degradation of acetone and higher ketones via carboxylation by newly isolated denitrifying bacteria. AB - Five strains of Gram-negative denitrifying bacteria that used various ketones as sole carbon and energy sources were isolated from activated sludge from a municipal sewage plant. Three strains are related to the genus Pseudomonas; two non-motile species have not yet been affiliated. All strains grew well with ketones and fatty acids (C2 to C7), but sugars were seldom utilized. The physiology of anaerobic acetone degradation was studied with strain BunN, which was originally enriched with butanone. Bicarbonate was essential for growth with acetone under anaerobic and aerobic conditions, but not if acetate or 3 hydroxybutyrate were used as substrates. An apparent Ks value of 5.6 mM bicarbonate was determined for growth with acetone in batch culture. The molar growth yield was 24.8-29.8 g dry cell matter (mol acetone consumed)-1, with nitrate as the electron acceptor in batch culture; it varied slightly with the extent of poly-beta-hydroxybutyric acid (PHB) formation. During growth with acetone, 14CO2 was incorporated mainly into the C-1 atom of the monomers of the storage polymer PHB. With 3-hydroxybutyrate as substrate, 14CO2 incorporation into PHB was negligible. The results provide evidence that acetone is channelled into the intermediary metabolism of this strain via carboxylation to acetoacetate. PMID- 2557379 TI - Biological response modifier enhances the activity of natural killer cell against human cytomegalovirus-infected cells. AB - Peripheral blood lymphocytes (PBL) from two human cytomegalovirus (CMV) seronegative donors and eight CMV-seropositive donors were cultured for 3 days with or without the biological response modifier OK-432 and examined for lysis of K562 cells and CMV-infected MRC-5 cells. OK-432-stimulated PBL exhibited significantly greater natural killer (NK) activity than did unstimulated PBL. There was no difference in activity of NK cells in PBL prepared from CMV seronegative and -seropositive donors. Antibody-complement depletion studies suggested that OK-432-stimulated NK activity was associated with Leu-7-positive cells. The ability of OK-432 to sustain the NK activity in PBL was decreased when the CD4-positive population of lymphocytes was eliminated by antibody-complement depletion prior to OK-432 stimulation. The ability of OK-432 to sustain the NK activity of PBL was also significantly decreased in the presence of monoclonal antibody against recombinant human interleukin-2. The results suggest that the activity of human NK cells against K562 and CMV-infected MRC-5 target cells can be sustained in vitro by OK-432-stimulated T-helper cells and that the effect of the T-helper cells is mediated, at least in part, by interleukin-2. PMID- 2557380 TI - Active replication of human immunodeficiency virus type 1 by peripheral blood mononuclear cells following coincubation with herpes viruses. AB - Patients with acquired immunodeficiency syndrome (AIDS) commonly suffer from opportunistic infections associated with members of the herpes virus family. To investigate whether certain of these other viruses might have an effect on the ability of the human immunodeficiency virus type 1 (HIV-1) to replicate, we coincubated peripheral blood mononuclear cells (PBMC) from nine HIV-1 seropositive donors with live preparations of various herpes viruses. In seven of nine cases, exposure of PBMC to preparations of either HSV-1, HSV-2, or CMV stimulated the cells to become active producers of HIV-1, as determined by reverse transcriptase activity and by the presence of infectious progeny virus. This increased production of HIV-1 particles appeared to be a consequence of mitogenic proliferation and of herpes virus-encoded transacting factors. These results supplement earlier findings on the molecular activation of the HIV-1 genome by both HSV and CMV genetic elements and point to a possible role for these viruses in the pathogenesis and ultimate clinical outcome of HIV-1 infections. PMID- 2557381 TI - Trans-regulation and differential cell specificity of human papillomavirus types 16, 18, and 11 cis-acting elements. AB - The noncoding region (ncr) of human papillomavirus (HPV) types 16, 18, and 11 contains promoter and/or enhancer function. We have localized the sequence containing the constitutive enhancers of HPV types 16, 18, and 11 to 315, 230, and 213 bp fragments, respectively, for comparative studies. The region of homology shared between the enhancers of the three viruses is limited to the sequence ATTTTTGGCTT, which is also present in the ncr of HPV 6b and 33. We have also examined the enhancer activity of the HPV ncrs in three human cervical carcinoma cell lines, one noncervical human carcinoma cell line, and one monkey kidney established cell line. We observed cell-specific differences in the constitutive expression of the enhancers in the various cell lines. The conditional enhancer activity of the ncr of the viruses is increased in trans by the E2 gene product of HPV 16. Transactivation by E2 is mediated through the E2 binding motif on HPV enhancer plasmids with a heterologous but not with a homologous promoter. Our preliminary studies also indicate a repressor function for the E7 gene of HPV 16. PMID- 2557382 TI - Detection of anti rotavirus coproantibodies by immunoblotting technique. AB - IgA and IgG coproantibodies to individual simian rotavirus (SA 11) structural polypeptides were detected in healthy infants in nursery homes. The number of immunoblottable peptides differed from individual to individual. Coproantibodies were also detected at the convalescent stage of rotavirus infection in two patients but not during the acute stage. This method is useful for confirming the diagnosis of rotavirus infection serologically without the need for paired sera. PMID- 2557383 TI - Passive protection of mice, goats, and monkeys against Japanese encephalitis with monoclonal antibodies. AB - Six monoclonal antibodies (McAbs) against Japanese encephalitis virus (JEV) were tested for passive protection in JEV-infected mice, goats, and rhesus monkeys. mG9 and nG2 had no protective effect; mG3 and 2D2 had some protective effect, but not sufficient to be of therapeutic significance; and 2H4 and 2F2 had excellent protective efficacy in mice even 120 hr after infection when most of the mice in the virus control group were sick. The mixture of 2H4, 2F2, mC3 (M-McAb), and their F(ab')2 fragments showed excellent protection in mice, goats, and monkeys and was safe. The protective effects of McAbs correlated with their neutralization titers, but cytotoxicity-mediated activities also played a role in protection. PMID- 2557384 TI - Complement-independent neutralising monoclonal antibody with differential reactivity for strains of human cytomegalovirus. AB - A mouse monoclonal antibody with complement-independent neutralising activity against cytomegalovirus (CMV) and reactive with the 86 kilodalton (kDa) viral glycoprotein H is described. Neutralisation tests against a range of different strains of CMV showed significant crossreactivity, but clear differences were evident between the two prototype viruses AD169 and Davis, and particularly between AD169 and several low-passage recent clinical isolates; CMV present in urine was neutralised weakly if at all. PMID- 2557386 TI - Detection of human cytomegalovirus immediate early antigen in leukocytes as a marker of viremia in immunocompromised patients. AB - Peripheral blood polymorphonuclear (PMN) cells from 35 immunocompromised patients (22 heart transplant recipients and 13 AIDS patients) and four normal subjects were tested for the presence of human cytomegalovirus (HCMV) immediate early antigen (IEA) (antigenemia) by indirect immunofluorescence (IFA) and IEA-specific monoclonal antibodies (MAb). PMN samples were tested in parallel for HCMV isolation (viremia) by using MAb to viral early antigens (EA) and the IFA technique 24-48 hr after inoculation onto human fibroblast monolayers. HCMV was isolated from 26 of 83 PMN samples examined: of these, 25 were also positive for HCMV IEA (96% sensitivity). Seven additional PMN samples negative for viral isolation resulted IEA-positive (87.7% specificity). Six of the seven discordant samples were taken from four patients during ganciclovir treatment. The transitory dissociation between positive HCMV antigenemia and negative viremia during antiviral treatment was followed, at the end of the therapy, either by virus clearance and disappearance of IEA-positive PMNs (one patient) or by reappearance of viremia (three patients). Among concordant positive samples, a significant correlation was observed between the number of IEA-positive PMN leukocytes and EA-positive nuclei of infected fibroblasts, when the same number of PMNs were used for both tests. PMID- 2557385 TI - Protective effect of a low-dose of cyclophosphamide in experimental infection of guinea pigs with Junin virus. AB - Administration of cyclophosphamide (CY) to guinea pigs infected with a lethal strain of Junin virus (JV) delayed the time of death, with survival of a small number of animals. Virological studies showed a temporary decrease of virus concentration in blood and viscera shortly after the CY injection. In the pathological study no differences were found in the organic lesions present in CY treated and nontreated animals, with the exception of the pulmonary alterations. In CY-treated guinea pigs the lungs appeared almost normal, but in the control, nontreated animals severe alterations with the pattern of the "respiratory distress syndrome of the adult" were consistently present. In in vitro experiments, incorporation of serum collected from guinea pigs injected 30 minutes before exsanguination with CY to cell cultures, infected with JV, prevented virus replication. On the basis of these results it is suggested that the delay of time of death and eventual survival of CY-treated guinea pigs after JV infection depends on a direct antiviral effect of the drug rather than on its known immunosuppressive action. In addition, the absence of pulmonary alterations in CY-treated animals was tentatively considered to be dependent on the marked polymorphonuclear leukocyte depletion induced by the drug. PMID- 2557387 TI - Interactions of water-soluble porphyrins with hexadeoxyribonucleotides: resonance raman, UV-visible and 1H NMR studies. AB - The interactions of the water-soluble porphyrins M(TMpy-P4) [M = H2, Cu(II), Ni(II), and Co(III); TMpy-P4 = tetrakis(4-N-methylpyridyl)porphyrinato ion], with the hexadeoxyribonucleotides d(CGTACG)2, d(TACGTA)2, d(GCATGC)2, d(TGTGCA)2, and d(CTATAG)2 have been investigated by resonance Raman and/or UV-visible spectroscopy. The results indicate that all hexamers containing the 5'CG3' as well as the 5'GC3' site, and also the mismatched hexamer d(TGTGCA)2, are capable of intercalating the H2, Cu(II) and Ni(II) porphyrins. 1H nuclear magnetic resonance spectra of d(CGTACG)2 mixed with Cu(TMpy-P4) have provided further evidence for the intercalation. For the other cases, outside binding by localized electrostatic interaction is suggested. There is no evidence of groove binding to any of the hexamers. Possible reasons for different binding properties of long and short helices are discussed. PMID- 2557388 TI - Heme-peptide/protein interactions: the binding of heme octa and undecapeptides, and microperoxidase-8 and -11, to human serum albumin. AB - The interaction of the heme octa (MP-8) and undeca (MP-11) peptides derived from cytochrome c with lipidated human serum albumin (HSA) has been investigated in aqueous solution. It is demonstrated that complex formation occurs in each case with a 1:1 stoichiometry. CN- binding has been used to investigate the accessibility of the heme in each complex by comparison with CN- interaction with methemalbumin. A preliminary study of the kinetics of the Fe3+MP-8/11 human serumalbumin (HSA) interaction demonstrates a clear ligand-size-related effect on mechanism of interaction--an ad hoc explanation of which is given in terms of HSA existing as two nonconverting conformers in solution. PMID- 2557389 TI - Preparation and spectral characterization of the heme d1.apomyoglobin complex: an unusual protein environment for the substrate-binding heme of Pseudomonas cytochrome oxidase. AB - The heme d1 prosthetic group isolated from Pseudomonas cytochrome oxidase combines with apomyoglobin to form a stable, optically well-defined complex. Addition of ferric heme d1 quenches apomyoglobin tryptophan fluorescence suggesting association in a 1:1 molar ratio. Optical absorption maxima for heme d1.apomyoglobin are at 629 and 429 nm before, and 632 and 458 nm after dithionite reduction; they are distinct from those of heme d1 in aqueous solution but more similar to those unobscured by heme c in Pseudomonas cytochrome oxidase. Cyanide, carbon monoxide and imidazole alter the spectrum of heme d1.apomyoglobin demonstrating axial coordination to heme d1 by exogeneous ligands. The cyanide induced optical difference spectra exhibit isosbestic points, and a Scatchard like analysis yields a linear plot with an apparent dissociation constant of 4.2 X 10(-5) M. However, carbon monoxide induces two absorption spectra with Soret maxima at 454 or 467 nm, and this duplicity, along with a shoulder that correlates with the latter before binding, suggests multiple carbon monoxide and possibly heme d1 orientations within the globin. The 50-fold reduction in cyanide affinity over myoglobin is more consistent with altered heme pocket interactions than the intrinsic electronic differences between the two hemes. However, stability of the heme d1.apomyoglobin complex is verified further by the inability to separate heme d1 from globin during dialysis and column chromatography in excess cyanide or imidazole. This stability, together with a comparison between spectra of ligand-free and -bound derivatives of heme d1 apomyoglobin and heme d1 in solution, implies that the prosthetic group is coordinated in the heme pocket through a protein-donated, strong-field ligand. Furthermore, the visible spectrum of heme d1.apomyoglobin varies minimally with ligand exchange, in contrast to the Soret, which suggests that much spectral information concerning heme d1 coordination in the oxidase is lost by interference from heme c absorption bands. A comparison of the absorption spectra of heme d1.apomyoglobin and Pseudomonas cytochrome oxidase, together with a critical examination of the previous axial ligand assignments from magnetic resonance techniques in the latter, implies that it is premature to accept the assignment of bishistidine heme d1 coordination in oxidized, ligand-free oxidase and other iron-isobacteriochlorin-containing enzymes. PMID- 2557391 TI - Localization of tetrodotoxin-sensitive field potentials of CA1 pyramidal cells in the rat hippocampus. AB - 1. The role of tetrodotoxin (TTX)-sensitive (Na+) channels in the generation of antidromic and orthodromic field potentials of the CA1 pyramidal cell population was examined by local application of TTX in the in vitro rat hippocampal slice preparation. 2. The sensitivity of alvear (antidromic) and stratum oriens (SO) evoked potentials to TTX application (10-100 microM) was tested in stratum pyramidale and over the entire extent of pyramidal cell apical dendrites in stratum radiatum. Stratum radiatum (SR)-evoked potentials were examined at the level of pyramidal cell bodies and over the proximal 200 microns of the apical dendritic region. 3. Pressure application of TTX confined to stratum pyramidale or regions of stratum radiatum selectively blocked the negative component of antidromic and SO-evoked population discharge in the cell body layer and over the initial 200 microns of stratum radiatum. 4. SR stimulation evoked a complex field potential in the proximal stratum radiatum (less than 150 microns) composed of at least three components: 1) A short-duration (approximately 3 ms) negativity of shorter peak latency than the population spike recorded simultaneously in stratum pyramidale. This potential was highly sensitive to TTX and appeared to be instrumental in the generation of the cell body population response. 2) A long duration negativity (approximately 20 ms) evoked at stimulation strengths that were subthreshold for both the short-duration negativity in proximal stratum radiatum and the cell body population spike. Although apparently less sensitive to TTX, this potential was reduced in amplitude with repeated TTX application; and 3) a slow (approximately 12 ms) positive-going potential that was only observed after eliminating all TTX-sensitive conductance mechanisms in the proximal stratum radiatum. 5. The latency difference between the SR-evoked short duration negativity of proximal stratum radiatum and the population spike in stratum pyramidale decreased or reversed during the course of multiple discharge induced by the addition of bicuculline or picrotoxin (5-10 microM) to the perfusate. 6. These data indicate the presence of TTX-sensitive presumed Na+ channels over the initial 200 microns of pyramidal cell apical dendrites capable of supporting active conduction of population discharge evoked by antidromic or SO stimulation. The sensitivity of SR-evoked potentials to TTX suggests that a synaptic potential generated in the distal apical dendrites is capable of triggering both a slow active depolarization and a fast spike-like discharge in the proximal apical dendritic region.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557390 TI - Comparative effects of F1 and P1 fractions obtained from a Klebsiella pneumoniae glycoproteic extract (RU 41740) on polymorphonuclear leukocytes. AB - RU 41740 is a glycoprotein extract from Klebsiella pneumoniae described as a macromolecular aggregation of a lipopolysaccharide (LPS)-associated protein (F1 fraction) and a glycoproteic complex (P1 fraction). The human polymorphonuclear (PMN) response was studied after incubation of the cells in the presence of RU 41740, F1 and P1 fractions, or F1-P1 complex. Oxidative metabolism was assessed by chemiluminescence, O2 consumption, O2- generation, and degranulation by beta glucuronidase release. Results were compared to data obtained with a homologous LPS. RU 41740, F1 fraction, and F1-P1 complex increased the respiratory burst of PMNs stimulated by opsonized zymosan (OZ). N-formylmethionylleucylphenylalanine (fMLP), phorbol myristate acetate, or the calcium ionophore A23187. The beta glucuronidase release was stimulated by the same compounds when OZ or fMLP were used as stimuli. These effects were dose-dependent. In contrast, P1 fraction was inactive. Addition of polymyxin B resulted in a profound inhibition of both the F1 fraction and LPS activities but only in a partial inhibition of RU 41740 effects. These results strengthen the hypothesis that different biochemical pathways are involved in the enhancement of stimulated neutrophil functions by RU 41740. PMID- 2557393 TI - Pleomorphic adenoma: a case report and review of the literature. PMID- 2557394 TI - Needed: a trial of competence. PMID- 2557392 TI - Differential effects of extra- and intracellular anions on GABA-activated currents in bullfrog sensory neurons. AB - 1. Kinetic properties of gamma-aminobutyric acid (GABA)-gated inward and outward anion currents were investigated in the frog sensory neurons perfused internally and externally with various anions with the use of a rapid concentration-jump (termed as 'concentration-clamp') technique. 2. Extracellular Br- [( Br-]o) shifted the dose-response curves of GABA-induced inward anion currents to the left without affecting the maximum values, whereas [Cl-]o, [I-]o, [No3-]o, [HCOO ]o, and [CH3COO-]o altered the rate of desensitization differently without shifting the GABA dose-response curves, indicating that the kinetics of desensitization phase are affected differently by various extracellular anions. 3. [CH3COO-]o suppressed the maximum current of the dose-response curve of the GABA-induced inward ICl without affecting Kd. 4. Both activation and desensitization phases of GABA-induced ICl consisted of fast and slow components, respectively. [Br-]o, [I-]o, and [NO3-]o significantly prolonged the slow desensitization component, whereas both [HCOO-]o and [CH3COO-]o shortened it. The fast desensitization and the fast and slow activation components were also affected by these foreign anions. 5. GABA dose-response curves of inward currents carried by various intracellular anions (Cl-, Br-, NO3-, I-, SCN-, HCOO-, F-, CH3COO-, CH3CH2COO-, BrO3-, and ClO3-) while keeping a constant [Cl-]o had a constant Kd value but different saturating maximum currents. There were no marked differences among their current kinetics except in the case of SCN-, indicating that the current kinetics is not affected by replacing intracellular Cl- [( Cl ]i) with various foreign anions. 6. The configuration and amplitude of GABA-gated outward anion currents at a constant [Cl-]i reflected the extracellular action of individual anions on the anion-binding site of GABA receptor associated with the anion-selective channel. 7. The relative conductances of the various anions, calculated from the maximum peak currents in dose-response curves of the GABA induced inward anion currents at a constant [Cl-]o, was in the sequence: I- greater than Br- greater than or equal to NO3- greater than ClO3- greater than SCN- greater than or equal to Cl- greater than HCOO- greater than BrO3- greater than CH3COO- greater than F- greater than CH3CH2COO-.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557395 TI - Risk-taking propensity of nurses: ADN and BSN. AB - Nursing literature indicates that skills in decision making are inherently important to the modern nurse. The literature also reports a controversy between the associate degree nurse (ADN) and the bachelor degree nurse (BSN). This conflict relates to educational level and experience in terms of leadership and decision-making skills. All decisions involve risk. Therefore decisions in nursing should be influenced by the risk-taking propensity of the individual nurse. The literature shows a linkage between risk-taking propensity and both training and experience. The major purpose of the study was to attempt to correlate risk-taking propensity of nurses by level of education and experience. The study investigated whether differences exist between risk-taking propensity of recently graduated ADNs and BSNs without experience. In addition, the study attempted to determine any differences in risk-taking propensity of ADNs and BSNs with practical experience. The results were analyzed in terms of decision-making competency. PMID- 2557396 TI - Smoking practices among nursing students: a comparison of two studies. AB - Findings of numerous studies that have explored the smoking practices of nurses reveal a high incidence of smoking that is incongruent with the health beliefs of the profession. Nurses who smoke are less likely to teach or positively influence patients who smoke. They may even undermine health teaching efforts of other health professionals. Studies of smoking practices of nursing students also reveal high incidences of smoking. Included among the determinants of this practice are lack of knowledge of the health effects of smoking, the academic setting, and the role that nursing education may play. In this comparison of two independent studies similar in design, smoking rates were similar to that of the female population and to registered nurses. Students who smoke either started smoking or increased their smoking in nursing school. They knew the health hazards of smoking and most had tried to quit in the past. Challenging opportunities exist for nurse educators to study and implement strategies to prevent smoking initiation and encourage cessation among future nurses. PMID- 2557397 TI - Effects of calculator use on arithmetic and conceptual skills of nursing students. AB - The purpose of this study was to determine the effect of the use of calculators on the responses of undergraduate nursing students to items on a dosage calculations examination. The study sought to determine if calculator use had any influence on both the arithmetic and conceptual skills associated with the solving of calculations problems. The population consisted of all students enrolled in a pharmacology course in a baccalaureate nursing program. An experimental repeated measures design was used in which all participants completed two parallel forms of a calculations examination, one form with a calculator and one form without a calculator. Findings of the study revealed that there was significant difference in both arithmetic and conceptual skills ability using a calculator. Use of a calculator was associated with improved arithmetic skills performance and diminished conceptual skills performance. A major factor underlying both arithmetic skills and conceptual skills performance, independent of the calculator effect, was the pre-admission arithmetic skills ability of the subject. PMID- 2557398 TI - Student perceptions of the links between nursing and the liberal arts. AB - This article examines the extent to which nursing students are aware of the importance of liberal arts in their professional education. Based on a survey of nursing students and alumnae from a small, New York state college, the project further explored the channels through which they became aware of the significant links between these two aspects of their education. One finding is that nursing students give a relatively high weight to the importance of the liberal arts in their total learning process. The more important conclusion is that students, on the whole, believe that they have made the links between liberal arts and nursing themselves. To the extent that other agents help forge these links, the most important seem to be nursing labs and clinicals, other nursing students, and nursing texts and modules. Students perceive that nursing faculty help draw links more often than liberal arts faculty, but neither rank very high. PMID- 2557399 TI - Dimensions of leadership of assistant/associate deans in collegiate schools of nursing. AB - The purpose of this study was to discover whether there were differences in the perceptions of deans, assistant/associate deans, and faculty of leadership styles, initiating structure and consideration, and position power of assistant/associate deans in selected collegiate schools of nursing. In addition, the effect of leadership styles on performance as perceived by the three groups related to the position power of assistant/associate deans was also examined. Collegiate schools of nursing, identified as having a hierarchal organization structure with persons identified by the title "assistant/associate dean," were invited to participate. Data were collected from a sample of 36 collegiate nursing schools. The tools used were the Ohio State Leadership Behavior Description Questionnaire, and the Fiedler Position Power Scale. Three questions were posed. A multi-variate analysis of testing the three hypotheses showed a significant difference between the three groups' perceptions of initiating structure, consideration, and position power. A main effect for initiating structure was found in the deans' perceptions of position power. A main effect for consideration was found in the perception of the faculty sampled. PMID- 2557400 TI - Pathways to information literacy. PMID- 2557401 TI - Young scholars' program: an innovative educational recruitment strategy. PMID- 2557402 TI - Teaching foreign students. PMID- 2557403 TI - BSN courses for students via satellite. PMID- 2557404 TI - Effects of feeding fermentable carbohydrates on the cecal concentrations of minerals and their fluxes between the cecum and blood plasma in the rat. AB - This study was conducted to determine in rats to what extent fermentable carbohydrates alter the mineral composition of cecal contents and the absorption of the major cations. The carbohydrates studied were as follows: an oligosaccharide (lactulose, 10%); a soluble fiber (pectin, 10%); and an amylose rich starch, incompletely broken down in the small intestine (amylomaize starch, 25 or 50%). All of these carbohydrates elicited a marked enlargement of the cecum, a drop of cecal pH and an increase in the volatile fatty acids (VFA) pool. With the lactulose diet, the VFA concentration was the lowest, whereas VFA absorption was similar to that observed with the 10% pectin or 25% amylomaize diets. From comparisons between germfree and conventional rats adapted to a fiber free diet, it appears that VFA are required as counter anions to maintain high concentrations of cations, especially sodium. In conventional rats fed fermentable carbohydrates, sodium concentration in the cecal fluid was approximately 80 mM, except with the lactulose diet (49.5 mM), due to osmotic effects of lactulose. There was, compared to the fiber-free diet, an increase in the cecal concentrations of potassium, calcium and phosphate, but not of magnesium; nevertheless, the cecal pool of all of these minerals was considerably increased. Potassium absorption was increased by fermentable carbohydrates in the cecum, which also appears to be a major site of magnesium and calcium absorption. Thus, fermentable carbohydrates shift aborally the absorption of the major cations, and this point is especially interesting in regard to calcium, since an enhanced supply of calcium in the large bowel has been invoked for fiber effects on colonic carcinogenesis. PMID- 2557405 TI - Myofibrillar and nonmyofibrillar myocardial proteins of copper-deficient rats. AB - Myofibrillar and nonmyofibrillar proteins from hearts of copper-adequate (n = 9) and copper-deficient (n = 10) rats were compared. Male weanling Long-Evans rats were fed copper-deficient or copper-adequate diets for 9 wk. Twelve additional rats were fed similar diets and cardiac tissue was evaluated by transmission electron microscopy. Ventricular myocytes were glycerinated and homogenized in 0.1 M KCl and 1.5% Triton X-100, and suspensions were centrifuged at 1100 x g. The supernatant was removed and designated Triton X-100-soluble non-myofibrillar protein, and the pellet was resuspended and recentrifuged several times to obtain myofibrillar protein. Sodium dodecyl sulfate--polyacrylamide gel electrophoresis (SDS-PAGE) analysis was conducted on both protein fractions. Densitometer scans of SDS-PAGE pherograms of myofibrillar protein revealed no significant difference between copper-adequate and copper-deficient groups. Similar analysis of nonmyofibrillar protein revealed a consistent decrease or diminished level of a 23-kDa polypeptide among copper-deficient rat hearts. These results may be consistent with the findings that demonstrated fragementation of mitochondrial cristae and an increased area occupied by mitochondria in copper-deficient rat hearts. PMID- 2557406 TI - Lactose protects against estrogen-induced pigment gallstones in hamsters fed nutritionally adequate purified diets. AB - To evaluate the impact of dietary factors on gallstone induction in hamsters, male Syrian hamsters were fed for 2-8 wk purified diets that varied in type and amount of simple sugar (glucose vs. lactose, 17.5-72%), fat (2-5%), fiber (0-15%) and estrogen (0 or 300 micrograms/kg diet). Plasma and liver cholesterol and plasma triglycerides were measured, daily weight gain was determined, cecal weights were obtained, and gallbladder bile was scored by light microscopy and analyzed chemically for its lithogenicity and gallstone incidence. Lactose reduced plasma lipids, especially triglycerides, and hepatic cholesterol accumulation, and maintained a lower biliary cholesterol concentration. When fed at 30% or more, lactose reduced weight gain, increased cecal volume 2- to 4-fold and prevented gallstone formation. Diarrhea and death from 'wet tail' was associated with gallstones and was frequent in hamsters fed glucose without fiber, but its incidence was essentially eliminated by rice flour plus fiber or lactose. Under these experimental conditions of time and diet, estrogen supplementation was required for the formation of gallstones. These appeared to be pigment stones containing a minimal amount of cholesterol. In summary, pigment gallstones were induced in less than 8 wk in hamsters fed estrogen-supplemented purified diets. Lactose feeding improved lipid metabolism and reduced gallstone formation, apparently through its impact on large bowel metabolism. PMID- 2557408 TI - Resistance of Escherichia coli to nourseothricin (streptothricin): reduced penetrability of the cell wall as an additional, possibly unspecific mechanism. AB - The resistance of E. coli strains to the antibiotic nourseothricin is known to be caused by an acetyltransferase acetylating the beta-lysine chain of the antibiotic. In addition, most of the resistant strains exhibit reduced penetrability of the outer membrane, presumably caused by a reduced amount of available negative charges. This was shown using crystal violet, Congo red, or the hydrophobic antibiotic novobiocin as indicators. PMID- 2557407 TI - Neuropeptide Y (NPY) innervation of the ovine pineal gland. AB - A dense network of neuropeptide Y (NPY)-like immunoreactive (NPY-LI) fibres was revealed in the ovine pineal gland at the light microscope level. The dorsal and peripheral regions of the gland contained the most dense concentration of NPY-LI fibres with relatively few fibres in the mid-region and almost none in the pineal stalk. The effect of NPY in conjunction with isoproterenol (ISO) on cyclic AMP (cAMP) accumulation and noradrenaline (NA) on melatonin synthesis was investigated using in vitro techniques. NPY had no effect on the stimulation of cAMP or melatonin synthesis by the adrenergic agonists. PMID- 2557409 TI - Depressor and natriuretic effects of M&B 22,948, a guanosine cyclic 3',5' monophosphate-selective phosphodiesterase inhibitor. AB - We examined the effects of an acute infusion of M&B 22,948 (2-o-propoxyphenyl-8 azapurin-6-one), a (cGMP)-selective phosphodiesterase inhibitor, on mean arterial pressure (MAP) and urinary sodium excretion in anesthetized rats. M&B 22,948 (at doses of 0.34-2.72 mg/kg/min for 30 min) lowered MAP in a dose-dependent manner, with a 60 mm Hg fall in pressure produced at the highest dose. Despite large decreases in MAP, a profound natriuresis was observed at all doses. Plasma concentrations of cGMP increased in parallel with the depressor action of M&B 22,948, whereas increases in the urinary excretion of cGMP temporally correlated with the natriuresis. The concentration of cyclic AMP in plasma increased transiently in rats treated with M&B 22,948 but the urinary excretion of cyclic AMP was not elevated in these animals. Because changes in cGMP correlated with the physiological effects of M&B 22,948, and the increase in cyclic AMP did not, it is likely that the depressor and natriuretic actions of M&B 22,948 are mediated by increases in cGMP. M&B 22,948 administered chronically at an oral dose of 200 mg/kg/day normalized MAP in spontaneously hypertensive rats; whereas MAP in vehicle-treated spontaneously hypertensive rats remained at hypertensive levels. cGMP-selective phosphodiesterase inhibitors (like M&B 22,948) could be more effective antihypertensive drugs than currently available vasodilators because, when administered acutely, M&B 22,948 simultaneously lowers blood pressure and promotes sodium excretion in the anesthetized rat. PMID- 2557410 TI - Dissociation between muscarinic receptor-mediated inhibition of adenylate cyclase and autoreceptor inhibition of [3H] acetylcholine release in rat hippocampus. AB - Activation of muscarinic cholinergic receptors (mAChRs) in the central nervous system reduces the catalytic activity of membrane-bound adenylate cyclase and attenuates depolarization-dependent release of acetylcholine (ACh). Inasmuch as reports have indicated that these mAChR-mediated responses exhibit pharmacological profiles similar to the M2 subclass of mAChR, the present studies were undertaken to ascertain whether attenuation of presynaptic adenylate cyclase activity [and concurrent reduction of intraneuronal cyclic AMP (cAMP) levels] underlies mAChR-mediated autoinhibition of electrically evoked ACh release. In [3H]choline-prelabeled rat hippocampal slices, the mAChR agonists oxotremorine (EC50 = 15 microM) and carbachol (EC50 = 80 microM) caused atropine-reversible inhibition of [3H]ACh release up to a maximum of 80% reduction. The rank order of potency for antagonist reversal of this inhibitory action (N-methylatropine = atropine greater than scopolamine much greater than pirenzepine) was generally consistent with an M2 mAChR-mediated response although pirenzepine was ineffective up to 1 mM. Under these assay conditions, forskolin (1-10 microM) and 8-bromo-cAMP (30-300 microM) enhanced electrically evoked [3H]ACh release maximally by 50 to 60%; however, neither agent significantly reversed mAChR agonist-induced inhibition of [3H]ACh release. Additional studies were undertaken to determine the consequences of chemically uncoupling mAChR from their G protein adenylate cyclase effector system in this tissue. Whereas brief pretreatment with the sulfhydryl alkylating agent N-ethylmaleimide (30 microM) or pertussis toxin (1 microgram/ml) markedly attenuated carbachol inhibition of adenylate cyclase activity in hippocampal tissue, there was no concurrent reduction of carbachol inhibited [3H] ACh release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557411 TI - Sympathoadrenal, cardiovascular and blood gas responses to highly selective mu and delta opioid peptides. AB - The relative importance of mu and delta opioid receptors in brain regulation of sympathoadrenal, cardiovascular and respiratory function was investigated using highly selective mu and delta opioid peptide analogs. Groups of conscious rats received i.c.v. injections of either the mu-selective agonist, [D-Ala2, MePhe4, Gly-ol5]enkephalin (DAMGO) or the delta-selective agonist, [D-Pen2, D Pen5]enkephalin (DPDPE). Blood pressure and heart rate were recorded continuously via a chronic catheter in the carotid artery, and arterial blood samples were taken at intervals through the same catheter for determination of blood pH, pCO2, pO2 and plasma catecholamine concentrations. Both DAMGO and DPDPE increased plasma catecholamine levels and blood pressure in a dose-related manner. The slopes of the dose-response lines were parallel, but the delta compound was about 250 times less potent than DAMGO. Only the highest dose of 5 nmol of DAMGO caused a significant bradycardia, mediated by parasympathetic (vagal) activation. DAMGO and DPDPE also induced dose-dependent acidosis, with DAMGO again being much more potent than DPDPE. The effects of both DAMGO and DPDPE on plasma catecholamines, blood pressure and blood gases were antagonized by a mu-selective dose of naloxone (0.4 mg/kg i.a.). Intracerebroventricular administration of the delta selective antagonist, ICI 174,864, only partially attenuated sympathoadrenal and blood gas responses to DAMGO or DPDPE. The pressor responses to DAMGO or DPDPE were resistant to antagonism by ICI 174,864. These results indicate that brain opioid receptors regulating autonomic outflow, cardiovascular and respiratory function are mainly of the mu type, although a delta opioid system may contribute to sympathoadrenal and respiratory effects of opioids. PMID- 2557412 TI - Comparative electroencephalographic and behavioral effects of phencyclidine, (+) SKF-10,047 and MK-801 in rats. AB - Female Sprague-Dawley rats prepared with chronic i.v. cannulas and/or cerebrocortical electrodes were administered sequentially increasing doses of phencyclidine (PCP, 0.1-6.4 mg/kg/injection), (+)-SKF-10,047 [(+)-N allynormetazocine] (0.4-25.6 mg/kg/injection) or MK-801 [(+)-5-methyl-10,11 dihydro-5H-dibenzo[a,d]-cyclohepten-5,10-imine maleate] (0.01-0.64 mg/kg/injection). Effects on overt behavior, cortical EEG power spectra, locomotor activity and rotarod performance were assessed. Quantitative EEG spectral parameters (peak, mean and edge frequency; total and relative power; time domain descriptors mobility and complexity) were analyzed from the global frequency range of 1 to 50 Hz. Increasing doses of each drug produced increases in EEG spectra power from 1 to 50 Hz which was associated with a slowing of the peak frequency. PCP and MK-801 produced decreases in the mean frequency, mobility and edge frequency whereas (+)-SKF-10,047 produced increases in these spectral parameters. Moreover, (+)-SKF-10,047 increased complexity whereas MK-801 decreased complexity and PCP did not change this parameter. Total spectral power from 20 to 50 Hz was increased by (+)-SKF-10,047 and PCP, but was not changed by MK-801. Each drug increased spontaneous locomotor activity. At the highest doses, PCP and MK-801 decreased activity whereas (+)-SKF-10,047 was lethal. Each drug disrupted rotarod performance. The rank order of potency for each effect was: MK 801 greater than PCP greater than (+)-SKF-10,047. The data indicate that subtle differences in the effects of these drugs can be detected using EEG power spectral analysis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557413 TI - Differential effects of phorbol ester on prefrontal cortex and striatal dopamine terminals: dependence on rate and duration of stimulation. AB - Compared to the nigrostriatal dopamine (DA) neurons, the mesocortical DA neurons projecting to the prefrontal cortex (PFC) are able to sustain higher levels of release when driven at high stimulation frequencies. The effect of a well known activator of protein kinase C (PKC), 4-beta-phorbol-12, 13-dibutyrate (PDBu), were compared on PFC and striatal DA terminals. DA release was monitored from slices of the rabbit PFC and striatum obtained from the same animal. The PKC activator, PDBu (30-1000 nM) enhanced the stimulation-evoked release (SER) of DA from PFC and striatum. The magnitude of the facilitation of DA release produced by PDBu was much greater from the PFC than from the striatum. In the striatum, PDBu produced a bell-shaped dose-response curve, i.e., 0.03 and 1 microM PDBu enhanced SER of DA by 25%, whereas 0.1 and 0.3 microM PDBu enhanced DA release by 60 and 100%, respectively (1 Hz, 120 pulses). In the PFC, 0.03 microM enhanced the SER of DA by 70% and 1 microM by 250% (1 Hz, 120 pulses). In addition, in the PFC, PDBu enhance the basal release of DA (+65% at 1 microM); this effect was not seen in the striatum. The inactive isomer, 4-alpha-phorbol-12, 13-dibutyrate (0.03-1 microM) failed to increase the SER and the basal release of DA from PFC or striatum. The SER of DA was dependent on the rate and duration of stimulation. However, under all conditions of stimulation studied DA release from PFC was always greater than from the striatum.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557414 TI - Antiarrhythmic properties of tetrodotoxin against occlusion-induced arrhythmias in the rat: a novel approach to the study of the antiarrhythmic effects of ventricular sodium channel blockade. AB - Blockade of ventricular sodium conductance (gNa) is believed to play an important role in the beneficial antiarrhythmic effects of class I antiarrhythmic agents. The present study was undertaken to examine the importance of ventricular gNa blockade by assessing the antiarrhythmic profile of tetrodotoxin (TTX), a selective sodium channel blocker. Experiments were performed in pentobarbital anesthetized and artificially ventilated rats. Two doses of TTX were tested for antiarrhythmic action: a low dose (low TTX, 10 micrograms/kg of bolus + infusion of 10 micrograms/kg/hr) which blocked only neuronal activity, and a high dose (TTXh, 50 micrograms/kg of bolus + infusion of 50 micrograms/kg/hr) which also produced signs of ventricular gNa blockade in normal hearts. To control for the decreases in blood pressure and heart rate caused by TTX, hexamethonium, nitroprusside and propranolol were also used. Only TTXh possessed antiarrhythmic activity in rats subjected to myocardial ischemia (produced by ligation of the left anterior descending coronary artery). Arrhythmia scores (mean, n = 9) were: saline, 3.8; hexamethonium, 3.8; nitroprusside, 3.2; nitroprusside + propranolol, 4.3; low TTX, 3.9; and TTXh, 0.9. Only TTXh reduced dV/dt max. of the action potential (recorded in vivo by means of 3 M KCl filled microelectrodes) as well as action potential height, and concomitantly prolonged the P-R and QRS intervals of normal hearts. In conclusion, our study demonstrated that drugs which produced hypotension, bradycardia and loss of autonomic function were not antiarrhythmic. On the other hand, the marked antiarrhythmic activity of TTXh appeared to depend upon ventricular gNa blockade. Thus, TTX provides a useful tool for examining the antiarrhythmic properties of ventricular gNa blockade. PMID- 2557415 TI - Effects of nicorandil on cytosolic calcium concentrations in quin2-loaded rat aortic vascular smooth muscle cells in primary culture. AB - We made use of quin2 microfluorometry to observe the effects of nicorandil (2 nicotinamidoethyl nitrate) on cytosolic Ca++ concentrations [( Ca++]i) in rat aortic vascular smooth muscle cells in primary culture. Regardless of whether cells were at rest, in a state of Ca++-depletion or at K+-depolarization, nicorandil rapidly and dose-dependently decreased [Ca++]i to the lower steady state level. Nicorandil dose-dependently inhibited norepinephrine-induced Ca++ transients in physiological salt solution containing 1 mM Ca++. Nicorandil accelerated the reduction of [Ca++]i observed when the cells were exposed to Ca++ free solution. When the cells were treated with nicorandil in Ca++-free solution, Ca++ transients induced by the first application of caffeine were little affected, but those induced by subsequent repetitive caffeine applications were reduced strongly and progressively. In contrast, pretreatment with nicorandil markedly inhibited Ca++ transients induced by the first application of norepinephrine, in Ca++-free solution. These effects of nicorandil on [Ca++]i and Ca++ transients were similar to those seen with nitroglycerin. The denitrated compound of nicorandil, N-(2-hydroxyethyl)nicotinamide, had no such effect. Thus, it is apparently the nitrate moiety of the chemical structure by which nicorandil actively and strongly reduces [Ca++]i in vascular smooth muscle cells. The reduction of [Ca++]i by nicorandil may result in a decrease in Ca++ in the norepinephrine-sensitive store; hence, the reduction of [Ca++]i elevation by norepinephrine. PMID- 2557416 TI - Renal handling of enalapril and enalaprilat: studies in the isolated red blood cell-perfused rat kidney. AB - An isolated recirculating or single pass red cell-perfused rat kidney preparation (IPK) was used to examine the differential handling of renal metabolites. In single pass experiments, enalapril was primarily metabolized to its polar, dicarboxylic acid metabolite, enalaprilat, and its fractional excretion (FE) was less than unity, suggesting net reabsorption. Its steady-state extraction ratio decreased from 0.3 to 0.2 at concentrations of 1.06 to 12.7 microM, due to a saturation of enzymes for esterolysis. Enalaprilat administered to the IPK was excreted into urine in a concentration-independent (0.41-35.3 microM) fashion, with FE values approximating unity, suggesting net filtration. Differences in handling were observed for enalaprilat, as a metabolite formed from enalapril and as an administered (preformed) species in the single pass IPK, when tracer concentrations of [14C]enalapril and [3H]enalaprilat were given simultaneously. A comparison made between steady-state extraction ratio Ess[mi] [generated metabolite]/glomerular filtration rate (GFR) and Ess[pmi] [preformed metabolite]/GFR, respectively, revealed a 2-fold difference. The finding suggests the presence of a barrier for entry of enalaprilat into the kidney. Or else, in absence of the barrier, the opposite would be observed, that is, Ess [pmi]/GFR greater than Ess [mi]/GFR because preformed enalaprilat, in contrast to generated enalaprilat, undergoes filtration and utilizes facilitative transport carriers at the basolateral membrane. In recirculating IPKs which received simultaneously a tracer bolus dose of [14C]enalapril and [3H]enalaprilat, the FE values for generated [14C]enalaprilat were high and variable, decreasing with perfusion time and exceeding those for preformed [3H]enalaprilat, which approached unity with perfusion time. The variable FE values for [14C]enalaprilat are due to time dependent contributions of circulating enalaprilat (which behaves identically to preformed enalaprilat) and the intrarenally generated enalaprilat. Hence, with renal drug metabolism, the conventional method of estimating urinary clearance (or Fe[mi]) for the metabolite [(total) excretion rate/midpoint plasma FE[mi] metabolite concentration] results in a greater metabolite clearance than that predicted from the administration of preformed metabolite. PMID- 2557417 TI - Contributions of various rat plasma peptidases to kinin hydrolysis. AB - The relative contribution of plasma carboxypeptidase N (kininase I), angiotensin converting enzyme (ACE) (kininase II), neutral endopeptidase 24.11 (enkephalinase A) and postproline cleaving enzyme to total kininase activity in rat plasma was determined by measuring bradykinin hydrolysis with and without various concentrations of inhibitors of these enzymes. We used DL-2-mercaptomethyl-3 guanidinoethyl-thiopropanoic acid to inhibit kininase I, enalaprilat for ACE, phosphoramidon for neutral endopeptidase 24.11 and N-benzyloxycarbonyl-Pro prolinal for postproline cleaving enzyme. Bradykinin was added to rat plasma and incubated at 37 degrees C. Kininase activity was evaluated based on the decrease in bradykinin during incubation. Bradykinin was measured by radioimmunoassay, using an antibody that recognizes its carboxyl group. Of the total plasma kininase activity, carboxypeptidase N was responsible for 11.0 +/- 2.5% (N = 5; P less than .05) and ACE for 46.8 +/- 1.5% (N = 5; P less than .001), whereas the contribution of neutral endopeptidase 24.11 and postproline cleaving enzyme turned out to be negligible. Of the kininase activity in rat plasma, 42% could not be explained by any of these four enzymes. We concluded that ACE is responsible for most of the kininase activity in rat plasma; carboxypeptidase N contributes to a slight degree. The fact that 42% of total plasma kininase activity could not be explained by any of the enzymes tested suggests that there are still other kininases in rat plasma which remain to be discovered. PMID- 2557418 TI - Electrophysiological effects of cocaine in the mesoaccumbens dopamine system: repeated administration. AB - Behavioral evidence indicates that the potent rewarding effects of cocaine are mediated, in part, by the mesoaccumbens dopamine (DA) system projecting from A10 DA cells in the ventral tegmental area (VTA) to the nucleus accumbens (NAc). Previous electrophysiological studies from our laboratory have indicated that cocaine (i.v.) exerts inhibitory effects on A10 DA neurons, due to enhanced stimulation by DA at DA autoreceptors are well as by activation of NAc-VTA feedback pathways. In the present experiments, extracellular single-unit recording and microiontophoretic techniques were used to determine the possible alterations in the mesoaccumbens DA system after repeated cocaine administration. Twice daily injections of cocaine (10 mg/kg i.p., 14 days) caused significant subsensitivity to the inhibitory effects of low i.v. doses of the DA agonist apomorphine in comparison to rats receiving similar treatments with saline or procaine. Iontophoretic application of DA to A10 DA neurons in rats treated repeatedly with cocaine (2X10 mg/kg, 14 days) also produced significantly less inhibition as compared to control rats. Cell population analysis of the VTA revealed that autoreceptor subsensitivity in cocaine-treated rats resulted in a significantly greater number of spontaneously active A10 DA neurons, and a significantly higher firing rate as compared to A10 DA neurons in control rats. In striking contrast to A10 DA cells, recordings from NAc neurons in cocaine treated rats (2X10 mg/kg, 14 days) indicated that these cells were supersensitive to the inhibitory effects of iontophoretic DA. Although the mechanism underlying such supersensitivity remains unclear, the increased sensitivity of postsynaptic NAc DA receptors combined with the subsensitivity of A10 DA autoreceptors could lead to greatly enhanced DA transmission and may help to explain some aspects of cocaine-induced behavioral sensitization. PMID- 2557419 TI - Guanyl-5'-yl-lmidodiphosphate regulation of ligand binding to LTD4 receptors on guinea pig lung membranes. AB - We investigated the mechanism of guanyl-5'-yl-imidodiphosphate (GppNHp) regulation of peptidoleukotrienes (LTs) and LT-antagonists binding to LTD4 receptors on guinea pig lung membranes (GPLMs). In saturation experiments, [3H]LTD4 saturable (maximum binding = 943 +/- 39 fmol/mg of protein) binding to GPLM was significantly (P less than .01) inhibited by GppNHp (60 nM, maximum binding = 446 +/- 113 fmol/mg of protein) in a concentration-dependent manner. No significant change in the affinity (Kd = 0.29 +/- 0.02 nM vs. 0.43 +/- 0.12 nM for control and treated GPLM, respectively) for [3H]LTD4 was observed. The binding affinity for the selective LTD4 antagonist ICI 198,615 (Ki = 0.13 +/- 0.04 nM) as determined by competition against [3H]LTD4, was not changed by GppNHp. Saturation analysis of [3H]ICI 198,615 binding confirmed that GppNHp did not change the apparent affinity or site-density for this ligand. In competition experiments against [3H]-ICI 198,615, GppNHp (1 microM) caused a significant (P less than .01) rightward shift of the inhibition by agonists (94-, 50- and 8-fold shifts for LTD4, LTE4 and YM-17690, respectively). In contrast, inhibition of [3H]ICI 198,615 by four LTD4 antagonists (ICI 198,615, 4-[5 cyclopentylcarbonylamino-1-[3-cyanobenzyl] indol-3-yl-methyl]3-methoxybenzoic acid, 4-[5-cyclopentylcarbonylamino-3-chloroindol-1-y-methyl]3-met hoxybenzoic acid and FPL55712) was not affected by GppNHp. Taken together the data suggest that LTD4 receptors are coupled to a G-protein that modulates the affinity of agonists but not antagonists binding. PMID- 2557420 TI - Defective modulation of noradrenergic neurotransmission by exogenous prostaglandins in aging spontaneously hypertensive rats. AB - Inhibition of cyclooxygenase enhances mesenteric vascular responses to periarterial (sympathetic) nerve stimulation (PNS) in 16-week-old spontaneously hypertensive rats (SHR), but not in 25-week-old SHR. In contrast, cyclooxygenase inhibition enhances mesenteric vascular responses to PNS similarly in 16- and 25 week-old Wistar-Kyoto normotensive rats (WKY). Thus, the modulation of noradrenergic neurotransmission by endogenous PGs becomes defective as SHR age, whereas in WKY this does not occur. The purpose of this study was to determine to what extent alterations in the concentrations of PGs and/or biological response to PGs contribute to this age/hypertension-related abnormality in SHR. All studies were conducted in the in situ autoperfused rat mesentery, and plasma levels of PGE2 and 6-keto-PGF1 alpha were determined by negative-ion, chemical ionization, gas chromatography-mass spectrometry after derivatization and clean up of samples by two thin-layer chromatographic steps. Base-line mesenteric venous plasma levels of PGs were similar in 16-week-old SHR vs. 16-week-old WKY; however, base-line levels of PGE2 were approximately 6-fold greater than base line levels of 6-keto-PGF1 alpha in both strains. PNS at 7 Hz approximately doubled mesenteric venous plasma levels of PGE2 in both 16-week-old SHR and WKY, but PNS did not increase levels of 6-keto-PGF1 alpha in either strain. Inasmuch as mesenteric venous plasma levels of PGE2 were responsive to PNS, the effect of aging on PGE2 levels was studied. In both strains, the base-line mesenteric venous plasma levels of PGE2 and the PNS-induced increase in PGE2 levels were similar in 16-week vs. 25-week-old animals. In 16-week-old SHR, infusions of PGE2, arachidonic acid and PGI2 directly into the mesenteric artery inhibited vascular responses to PNS. However, in 25-week-old SHR, even high doses of PGE2 or arachidonic acid failed to inhibit vascular responses to PNS, and the inhibitory potency of PGI2 was shifted 10-fold to the right compared to 16-week old SHR. In contrast, PGE2 and arachidonic acid had similar effects on neurotransmission in 25-week-old WKY compared to 16-week-old WKY, and aging had a lesser effect on the inhibitory potency of PGI2 (i.e., 3-fold rightward shift of the dose-response curve). Adenosine also inhibited vascular responses to PNS; however, the inhibitory potency of adenosine was only slightly and similarly affected by aging in SHR and WKY.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557421 TI - Effects of delta-9-tetrahydrocannabinol on sensory evoked hippocampal activity in the rat: principal components analysis and sequential dependency. AB - The effects of delta-9-tetrahydrocannabinol (delta-9-THC) were assessed on identified hippocampal sensory evoked potentials obtained from rats during performance of a two-tone discrimination task. Techniques which analyzed the trial-to-trial sequential and serial dependence underlying the variance in evoked potential amplitude were utilized. Waveforms of averaged tone-evoked potentials (AEPs) recorded from the outer molecular layer of the dentate gyrus (OM) were subjected to principal components analysis which revealed eight principal components accounting for 90.3% of the total variance in the set of OM AEP waveforms. Five of the eight components were altered significantly in comparison to vehicle injection sessions after administration of either a 1.0- or 2.0-mg/kg dose of delta-9-THC. These alterations accounted for the amplitude and latency changes in the OM AEP described in a previous report. In addition, delta-9-THC also disrupted the trial-to-trial sequential dependency of the OM AEPs. An important result showed that delta-9-THC selectively influenced the serial dependence of the OM AEP. These results implicate delta-9-THC as a potent disruptor of temporally specific information as it is processed by the hippocampus and suggest that such disruption may be the basis of delta-9-THC effects on memory processes in humans. PMID- 2557422 TI - Enhanced slow-pressor response to angiotensin II in spontaneously hypertensive rats. AB - Rapid-pressor and slow-pressor responses to angiotensin (ANG) II and norepinephrine (NE) in spontaneously hypertensive rats (SHR) and Wistar Kyoto control rats (WKY) were examined. All animals were treated from 4 wk of age with captopril (100 mg/kg/day in drinking water) to prevent development of hypertension so that changes in responsiveness could not be attributed to disparate base-line blood pressures or to hypertension-induced injury of the cardiovascular system. In 11-wk, conscious, unrestrained, captopril-treated rats, ANG II and NE induced rapid-pressor responses (i.e., a rapid increase in arterial blood pressure that reached a maximum within 10 min) that were of similar magnitude in SHR and WKY. In an additional group of 9-wk captopril-treated rats, both ANG II and NE caused slow-pressor responses (i.e., a slow increase in arterial blood pressure over 2 wk). Although the slow-pressor response to NE was similar in SHR versus WKY, the slow-pressor response to ANG II was much greater in SHR compared with WKY. Further studies were conducted in captopril-treated (from 4 wk of age) SHR and WKY to investigate whether the increased slow-pressor response to ANG II in SHR was mediated by an enhanced ability of ANG II to potentiate peripheral sympathetic neurotransmission, contract vascular smooth muscle, increase sympathetic tone to nonadrenal sites, release aldosterone, and/or reduce renal function. No evidence was found that supported a role for the aforementioned nonrenal actions of ANG II. However, 11-wk captopril-treated SHR were 10-fold more sensitive to the antidiuretic, antinatriuretic, and renal vascular effects of intrarenal infusions of ANG II compared with captopril treated WKY. Also, chronic (1 wk) intrarenal infusions of a very low dose of ANG II (1 ng/min) caused a marked slow-pressor response in 11-wk captopril-treated SHR but did not alter arterial blood pressure in WKY. We conclude that 1) the slow-pressor response to ANG II is greatly enhanced in SHR, 2) this enhancement is specific with respect to type of response (slow not rapid) and pressor agent (ANG II not NE), 3) a genetic defect underlies the increased slow-pressor response to ANG II in SHR, and 4) the enhanced slow-pressor response to ANG II contributes significantly to the pathophysiology of hypertension in SHR. Finally, the current studies are consistent with our working hypothesis that the kidneys mediate the enhanced slow-pressor response to ANG II in SHR. PMID- 2557423 TI - Different mechanisms of antagonism by methoctramine of two neuronal muscarinic receptor-mediated second messenger responses. AB - The allosteric effects and subtype selectivity of methoctramine on neuronal muscarinic receptors in N1E-115 cells and two different rat brain regions (cerebral cortex and striatum) were assessed. Saturation isotherms of [3H]N methylscopolamine binding, performed in N1E-115 cells and dissociated cerebral cortex, showed that methoctramine reduced the Bmax in a concentration-dependent manner. Furthermore, this compound slowed the rate of dissociation of bound [3H]N methylscopolamine in the same tissue preparations. Low concentrations of methoctramine (less than or equal to 1 microM) antagonized the M1-linked phosphoinositide response in N1E-115 cells and dissociated cerebral cortex in an apparent competitive mechanism. However, methoctramine exhibited noncompetitive effects at higher concentrations in N1E-115 cells. Observation of a similar effect in cerebrocortical cells was precluded since methoctramine by itself, at concentrations higher than 1 microM, stimulated inositol phosphate formation. The stimulatory effect of methoctramine on phosphoinositide hydrolysis was not blocked by atropine. A solely competitive mode of antagonism by methoctramine was observed for the inhibition of cAMP formation (a noncardiac-M2 coupled response) in N1E-115 cells and dissociated striatum. This antagonism was evident even at concentrations of methoctramine that noncompetitively antagonized the M1 response. Anomalously, methoctramine alone inhibited cAMP formation in dissociated striatum at concentrations of greater than or equal to 30 microM. Atropine was ineffective at blocking this effect. Methoctramine failed to demonstrate muscarinic receptor subtype selectively in blocking these two second messenger responses. This nonselectivity was supported by indirect binding experiments involving methoctramine and [3H]N-methylscopolamine. The data presented here demonstrate that methoctramine binds to a secondary site(s) associated with neuronal muscarinic receptors. Furthermore, methoctramine exhibits different mechanisms of antagonism and displays poor selectivity for the M1-linked phosphoinositide and noncardiac-M2 linked cAMP responses. PMID- 2557424 TI - Characterization of Ca2+ and K+ currents in the human Jurkat T cell line: effects of phytohaemagglutinin. AB - 1. Inward and outward currents were recorded in the human Jurkat T cell line using the whole-cell configuration of the patch-clamp technique. 2. The transient outward current was activated at membrane potentials positive to -60 mV. The activation time constant-voltage relationship decreased from 17 ms to 2 ms for membrane potentials ranging from -40 to +40 mV. The inactivation phase could be fitted by a single-exponential function and the inactivation time constant decreased from 250 ms to 150 ms for membrane potentials ranging from -20 to +100 mV. 3. The steady-state inactivation-voltage relationship showed a mid-point potential of -32 +/- 2.6 mV, and the slope factor was 10.8 +/- 1.8 mv (n = 3). 4. The calcium ionophore A23187 provoked a decrease in the amplitude of the outward current, suggesting a dependence of this current on the cytosolic concentration of Ca2+. 5. The K+ outward current was blocked by tetraethylammonium (TEA, Michaelis-Menten constant (Km), 6 mM) and by the calcium channel blockers Ni2+, Co2+, Mn2+ and Cd2+. 6. Forty per cent (n = 120) of the patched Jurkat cells displayed an inward current. In a physiological medium containing Ca2+ (2.2 mM), the inward current threshold voltage was -60 mV, the maximum current was observed at -40 mV and the zero current voltage was positive to +20 mV. At negative membrane potentials, the time required to reach 50% of the maximum amplitude was 60 ms and grew shorter with increasing depolarization, reaching a value of 5 ms at -5 mV. The inactivation of the inward current was very slow and the time constant varied from 1200 ms at -35 mV to approximately 250 ms for potentials positive to -10 mV. 7. The current availability had a value of one for potentials negative to -50 mV and zero for potentials positive to -15 mV. The mid-point potential was -31 +/- 3.4 mV and the slope factor was 3.3 +/- 0.2 mV (n = 3). 8. The inward channels were permeable to Sr2+, but were blocked by classical Ca2+ channel inhibitors such as Co2+, Mn2+ and Ni2+. 9. Phaseolus vulgaris phytohaemagglutinin (PHA), an inducer of interleukin-2 production in Jurkat cells, increased the inward current amplitude by 32 +/- 20% (n = 4). This increase was concomitant with a decrease (45 +/- 12%) in the amplitude of the outward current, but only when the current was carried by Ca2+.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557425 TI - Dihydropyridine-sensitive low-threshold calcium channels in isolated rat hypothalamic neurones. AB - 1. Low-voltage-activated Ca2+ channels which produce a transient inward current were studied in neurones freshly isolated from the ventromedial hypothalamic region of the rat. Membrane currents were recorded using a suction-pipette technique which allows for internal perfusion under a single-electrode voltage clamp. A concentration-jump technique was also used for rapid drug application. 2. In most cells superfused with 10 mM-Ca2+, a transient inward Ca2+ current was evoked by a step depolarization to potentials more positive than -65 mV from a holding potential of -100 mV. Such a low-threshold Ca2+ current could easily be separated from a high-threshold, steady type of Ca2+ current by selecting the holding and test potential levels, as well as by resistance to the wash-out during cell dialysis. 3. Activation and inactivation processes of the low threshold Ca2+ current were highly potential dependent at 20-22 degrees C. For a test potential change from -60 to +20 mV, the time to peak of the current decreased from 45 to 9 ms, and the time constant of the current decay decreased from 90 to 40 ms. The steady-state inactivation occurred at very negative potentials, reaching a 50% level at -93 mV. Recovery from inactivation showed a time constant between 2.63 and 0.94 s for a potential change from -80 to -120 mV. 4. The amplitude of the low-threshold Ca2+ current depended on the external Ca2+ concentration [( Ca2+]o), approaching saturation at 100 mM [Ca2+]o. Ba2+ substituted for Ca2+ reduced the current amplitude by 30-50% while Sr2+ produced no definite changes in the current amplitude. 5. The low-threshold Ca2+ current was blocked by various di- or trivalent cations in the sequence of La3+ greater than Zn2+ greater than Cd2+ greater than Ni2+ greater than Co2+. The corresponding apparent dissociation constants (KD) were 7 x 10(-7), 1 x 10(-4), 3 x 10(-4), 6 x 10(-4) and 3 x 10(-3) M. 6. Various organic Ca2+ antagonists were effective in blocking the low-threshold Ca2+ current in the following sequence: flunarizine greater than nicardipine greater than nifedipine greater than nimodipine greater than D600 (methoxyverapamil) greater than diltiazem. The corresponding KDs were 7 x 10(-7), 3.5 x 10(-6), 5 x 10(-6), 7 x 10(-6), 5 x 10( 5) and 7 x 10(-5) M. These Ca2+ antagonists induced a use-dependent decrease in the current amplitude.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557427 TI - Reduction by general anaesthetics of group Ia excitatory postsynaptic potentials and currents in the cat spinal cord. AB - 1. The effects of thiopentone and halothane on excitatory synaptic transmission at group Ia afferent synapses on lumbosacral motoneurones were studied in the anaesthetized or decerebrate cat. 2. Thiopentone (10 mg kg-1) infused on a background of light pentobarbitone anaesthesia caused a decrease in single-fibre monosynaptic group Ia excitatory postsynaptic potentials (EPSPs) of between 0 and 24%. A step increase in inspired halothane concentration in the range 0.7-0.9% produced a decrease in EPSP amplitude of between 0 and 31%. These effects were reversible when the anaesthetic level was reduced. 3. Fluctuation analysis of selected single-fibre group Ia EPSPs revealed that these effects could be accounted for by a decrease in the probability of occurrence of EPSPs of larger amplitude, and an increase in the probability of occurrence of EPSPs of smaller amplitude. The mean separation between discrete amplitudes was not altered by either anaesthetic agent. 4. EPSPs whose time course indicated a somatic site of origin were voltage clamped to study the effect of the anaesthetics on the time course of the synaptic currents. Neither thiopentone nor halothane produced a consistent effect on the time constant of decay of the current, although they both depressed its peak amplitude. 5. The results are interpreted as indicating a presynaptic site of action of both anaesthetics at the concentrations studied: the probability of release of neurotransmitter is reduced, without any detectable change in the mean duration of the postsynaptic conductance increase. These findings are discussed in relation to the mechanisms of action of anaesthetics on exocytosis and presynaptic inhibition. PMID- 2557426 TI - Voltage-dependent and calcium-dependent inactivation of calcium channel current in identified snail neurones. AB - 1. The dependence of Ca2+ current inactivation on membrane potential and intracellular Ca2+ concentration ([Ca2+]i) was studied in TEA-loaded, identified Helix neurones which possess a single population of high-voltage-activated Ca2+ channels. During prolonged depolarization, the Ca2+ current declined from its peak with two clearly distinct phases. The time course of its decay was readily fitted by a double-exponential function. 2. In double-pulse experiments, the relationship between the magnitude of the Ca2+ current and the amount of Ca2+ inactivation was not linear, and considerable inactivation was present, even when conditioning pulses were to levels of depolarization so great that Ca2+ currents were near zero. Similar results were obtained when external Ca2+ was replaced by Ba2+. 3. In double-pulse experiments, hyperpolarization during the interpulse interval served to reprime a portion of the inactivated Ca2+ current for subsequent activation. The extent of repriming increased with hyperpolarization, reaching a maximum between -130 and -150 mV. The effectiveness of repriming hyperpolarizations was considerably increased when Ca2+ was replaced by Ba2+. 4. A significant fraction of inactivated Ca2+ channels can be recovered during hyperpolarizing pulses lasting only milliseconds. If hyperpolarizing pulses were applied before substantial inactivation of Ca2+ current, Ca2+ channels remained available for activation despite considerable Ca2+ entry. 5. The relationship between [Ca2+]i and inactivation was investigated by quantitatively injecting Ca2+-buffered solutions into the cells. The time course of Ca2+ current inactivation was unchanged at free [Ca2+] between 1 x 10(-7) and 1 x 10(-5) M. From 1 x 10(-7) to 1 x 10(-9) M, inactivation became progressively slower, mainly due to a decrease of the amplitude ratio (fast/slow) of the two components of inactivation, which fell from about unity to near zero at 1 x 10(-9) M. In double pulse experiments, recovery from inactivation was enhanced in neurones that had been injected with Ca2+ chelator. 6. We conclude that inactivation of Ca2+ channels in these neurones depends on both [Ca2+]i and membrane potential. The voltage-dependent process may serve as a mechanism to quickly recover inactivated Ca2+ channels during repetitive firing despite considerable Ca2+ influx. 7. The results are discussed in the framework of a model which is based on two states of inactivation, INV and INCA, which represent different conformations of the inactivating substrate, and which are both reached from a lumped state of activation (A). Inactivation leads to high occupancy of INV during depolarization.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557428 TI - Effect of catecholamines on deformability of red cells from trout: relative roles of cyclic AMP and cell volume. AB - 1. In the presence of catecholamine the nucleated red blood cells of trout show a large increase in cell volume as a result of an accumulation of sodium and chloride due to activation of an amiloride-sensitive, cyclic AMP-dependent Na+-H+ exchanger allowing Na+ to enter in exchange for internal H+. 2. The activation of this cyclic AMP-dependent Na+-H+ exchange is considered to be involved in an adaptive response to hypoxia by increasing the oxygen-carrying capacity of erythrocytes. But cell swelling could increase resistance to blood flow and thus impair the expected physiological advantages for oxygen transport. The effect of catecholamine on the deformability properties of the red blood cells has been studied by measuring the rate at which blood flows through a Nucleopore filter (5 microM). 3. The results show that stimulation by catecholamine in fact increases the erythrocyte deformability, a response which must favour the supply of oxygen at the tissue level. 4. Hormonal stimulation increases the cellular cyclic AMP content (and cyclic AMP-dependent phosphorylation of cytoskeleton proteins could influence cell deformability) and the cell volume. It has been shown that when cellular cyclic AMP content is increased under conditions where the cell cannot swell, the erythrocyte becomes more rigid and not more deformable. Conversely the results show a systematic coincidence between cell swelling and deformability increase. The precise way in which volume change and deformability are interrelated needs more study. PMID- 2557429 TI - Two types of calcium currents in single smooth muscle cells from rat portal vein. AB - 1. Using the whole-cell recording mode of the patch-clamp technique, we investigated the calcium currents in isolated cells from rat portal vein in short term primary culture. 2. From a holding potential of -70 mV the cells presented two types of calcium currents with 5 mM-extracellular calcium: one type was activated by small depolarizations and inactivated quickly (fast calcium current), whereas the other required stronger depolarizations for activation and inactivated more slowly (slow calcium current). 3. Isradipine (PN 200-110) blocked the slow calcium current at concentrations 300 times lower than those used to block the fast inward current. The isradipine-induced inhibition was voltage-dependent for the slow calcium current and voltage-independent for the fast calcium current. 4. The slow calcium current was lost during internal perfusion with a 0.5 microM-Ca2+ containing solution, and during stimulation of the cell at high frequencies (0.1-0.2 Hz) within 5-10 min. The fast calcium current was unchanged under these experimental conditions. 5. Steady-state inactivation curves for both fast and slow calcium currents showed differences in their voltage dependence. Half-maximal and complete inactivations of the fast calcium current were obtained at -50 and -30 mV while those of the slow calcium current were obtained at -20 and +10 mV. 6. Studied with the two-pulse protocol, inactivation of the slow calcium current was dependent on both membrane potential and calcium influx while that of the fast calcium current appeared only dependent on membrane potential. 7. Two types of calcium currents, differing in potential dependence of inactivation, and in sensitivities to dihydropyridines, stimulation frequency and intracellular calcium concentration were identified in cultured smooth muscle cells isolated from portal vein. PMID- 2557430 TI - Calcium currents in the normal adult rat sympathetic neurone. AB - 1. The calcium currents evoked by membrane depolarization in the mature and intact rat sympathetic neurone have been studied at 37 degrees C using two electrode voltage-clamp analysis. 2. Under conditions that eliminate Na+ and K+ currents and 5 mM-external Ca2+, inward currents were observed that activated at about -30 mV and reached maximum amplitude between 0 and +10 mV with time-to-peak values (2.7-1.9 ms) decreasing with increasing membrane depolarization. Thereafter, calcium current (ICa) decayed to a virtually zero level with maintained depolarization. Two exponentials were required to describe the total inactivation process. The faster rate (tau = 29.3-17.6 ms) is ten times the slower rate and proved to be only slightly voltage-dependent. Double-pulse experiments gave a similar time course of turn-off. 3. No steady-state inactivation was removed at holding potentials between -40 and -70 mV and indirect data suggest that all the ICa was available at -50 mV. Within the -30 to -50 mV holding potential range no significant modifications either in the final amount of ICa inactivation or in the inactivation time constant values were detected. 4. After an initial 100 ms, recovery from inactivation followed a single-exponential process with a mean time constant value of 1.54 s at -50 mV. 5. The kinetics of ICa observed in this neurone were consistent with the existence of a single class of Ca2+ channels. For times up to 20 ms, ICa is described reasonably well by a Hodgkin-Huxley c2hc scheme. The activation time constant was 0.57 ms close to threshold and 0.29 ms at +30 mV. Deactivation occurred with a similar fast time course. The steady-state value of the variable c was evaluated in the -40 to +20 mV voltage range: 9.9 mV are required to change c infinity e-fold. 6. Following previous analyses, we have formulated a mathematical model which incorporates the present ICa kinetic equations with Hodgkin-Huxley-type gating mechanisms for INa, IA and IK(V) conductances. The Ca2+ load of the neurone proved to be basically an 'off' effect and to be governed by the duration of the action potential falling phase. The model is consistent with the experimental observations indicating that Ca2+ channels probably do not have an important direct electrical function in the sympathetic neurone spike at normal membrane potential levels.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557431 TI - Frequency-dependent depression of inhibition in guinea-pig neocortex in vitro by GABAB receptor feed-back on GABA release. AB - 1. The mechanisms involved in the lability of inhibition at higher frequencies of stimulation were investigated in the guinea-pig in vitro neocortical slice preparation by intracellular recording techniques. We attempted to test the possibility of a feedback depression of GABA on subsequent release. 2. At resting membrane potential (Em, -75.8 +/- 5.2 mV) stimulation of either the pial surface or subcortical white matter evoked a sequence of depolarizing and hyperpolarizing synaptic components in most neurones. An early hyperpolarizing component (IPSPA) was usually only obvious as a pronounced termination of the EPSP, followed by a later hyperpolarizing event (IPSPB). Current-voltage relationships revealed two different conductances of about 200 and 20 nS and reversal potentials of -73.0 +/ 4.4 and -88.6 +/- 6.1 mV for the early and late component, respectively. 3. The conductances of IPSPA and IPSPB were fairly stable at a stimulus frequency of 0.1 Hz. At frequencies between 0.5 and 2 Hz both IPSPs were attenuated with the second stimulus and after about five stimuli a steady state was reached. Concomitantly IPSPs were shortened. The average decrease in synaptic conductance between 0.1 and 1 Hz was 80% for the IPSPA and 60% for the IPSPB. At these frequencies the reversal potentials decreased by 5 and 2 mV, respectively; Em and input resistance (Rin) were not consistently affected. 4. The amplitudes of field potentials, action potentials and EPSPs of pyramidal cells were attenuated less than 10% at stimulus frequencies up to 1 Hz, suggesting that alterations in local circuits between the stimulation site and excitatory input onto inhibitory interneurones may play only a minor role in the frequency-dependent decay of IPSPs. 5. Localized application of GABA produced multiphasic responses. With low concentrations and application near the soma an early hyperpolarization prevailed followed by a depolarizing late component. Brief application of GABA at low frequencies induced constant responses; at higher frequencies, the responses sometimes declined. The current-voltage relationships of the two GABA responses were similar to each other and to the early IPSP. An apparently fivefold higher conductance was estimated at lower Ems, suggesting that the GABA response had a voltage sensitivity. The slope conductance of IPSPs was decreased by up to 50% for tens of seconds after postsynaptically detectable effects of GABA had dissipated. 6. Application of the GABA uptake inhibitor nipecotic acid (50-500 microM) reduced the conductance of both components of orthodromically evoked inhibition and shortened the IPSP at low frequencies, but had no additional effects at higher stimulation rates.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557433 TI - Pharmacological and anatomical separation of calcium currents in rat dentate granule neurones in vitro. AB - 1. Rat dentate granule neurones in hippocampal slices were voltage-clamped at 21 23 degrees C using CsCl-filled microelectrodes. The perfusate contained TTX and K+ channel blockers to isolate pharmacologically inward Ca2+ currents. 2. From hyperpolarized holding potentials of -65 to -85 mV, depolarizing test potentials to between -50 and -40 mV elicited a transient (100-200 ms) low-threshold (TLT) current which was also elicited from more depolarized holding potentials following hyperpolarizing voltage steps of -40 mV or greater. 3. Larger depolarizing steps from a hyperpolarized holding potential triggered a large (2-6 nA), transient high-threshold (THT) inward current, rapidly peaking and decaying over 500 ms, followed by a sustained inward current component. 4. At depolarized holding potentials (-50 to -20 mV), the THT current was apparently inactivated and a sustained high-threshold (SHT) inward current was evident during depolarizing voltage steps of 10 mV or more. 5. From hyperpolarized holding potentials with depolarizing voltage steps of 10-30 mV, most neurones demonstrated a small-amplitude, sustained low-threshold (SLT) inward current with similar characteristics to the SHT current. 6. Zero-Ca2+ perfusate or high concentrations of Ca2+ channel blockers (Cd2+, Mn2+ or Ni2+) diminished or abolished all inward currents. 7. Repetitive voltage step activation of each current at 0.5 Hz reduced the large THT current to less than 25% of an unconditioned control current, reduced the SHT current by 50%, but had little effect on the TLT current. 8. A low concentration of Cd2+ (50 microM) blocked the THT and SHT currents with little effect on the TLT current. Nimodipine (1 microM) attenuated the SHT current. Ni2+ (100 microM) selectively attenuated the TLT current. 9. In low-Ca2+ perfusate, high concentrations of Ca2+ (10-15 mM), focally applied to different parts of the neurone, increased the THT current when applied to the dendrites, the SHT current when applied to the soma and the TLT current at all locations. Conversely, in regular perfusate, Cd2+ (1-5 mM), focally applied to the dendrites decreased the THT current and somatic applications decreased the SHT current. The TLT current was diminished regardless of the site of Cd2+ application. 10. These results suggest the existence of three different Ca2+ currents in dentate granule cells separable by their activation and inactivation characteristics, pharmacology and site of initiation. PMID- 2557432 TI - Local inhibition of converting enzyme and vascular responses to angiotensin and bradykinin in the human forearm. AB - 1. The function of angiotensin converting enzyme was investigated in twenty-four healthy men. Forearm blood flow was measured under basal conditions and during administration of enalaprilat (a converting enzyme inhibitor) and/or peptide substrates of converting enzyme into the left brachial artery. Blood flow was compared in the two arms. 2. Enalaprilat had no effect on basal blood flow. The concentration of enalaprilat in venous blood from the control arm was low, and plasma renin activity was not increased, indicating that systemic inhibition of converting enzyme did not occur. 3. Effects of angiotensin and of bradykinin, administered intra-arterially, were limited to the infused arm. Enalaprilat (13 nmol min-1) inhibited converting enzyme in the infused arm, in which it caused approximately a 100-fold reduction in sensitivity to angiotensin I, while having no effect on the vasoconstriction caused by angiotensin II. Enalaprilat increased vasodilatation caused by bradykinin. 4. Aspirin, an inhibitor of cyclo-oxygenase, did not inhibit vasodilatation caused by bradykinin whether infused alone or with enalaprilat, indicating that these responses are not mediated by prostaglandins. 5. We conclude that under basal conditions neither conversion of angiotensin I to angiotensin II nor degradation of bradykinin determines resistance vessel tone in the human forearm. Converting enzyme may affect vascular tone in situations in which intravascular concentrations of peptides are increased over those present under basal conditions. PMID- 2557434 TI - Protection of hippocampal slices from young rats against anoxic transmission damage is due to better maintenance of ATP. AB - 1. Dentate granule cells in hippocampal slices from young rats (aged 30-40 days) are more resistant to damage from 10 min of anoxia than are granule cells from adult rats. The evoked population spike from these cells recovers to 78% of its pre-anoxic amplitude in young animals while in adult animals it shows only 4% recovery. This increased resistance is associated with higher levels of adenosine triphosphate (ATP) during the anoxic period. 2. When the duration of anoxia in slices from young animals is increased to 15 min, ATP falls to levels found in adult tissue after 10 min of anoxia. The dentate granule cells in slices from young animals show little recovery of the evoked response (19%) after such an exposure to anoxia. 3. When slices from young animals are subjected to 10 min of anoxia in low-glucose (2 mM) artificial cerebrospinal fluid, ATP levels fall to those found in adult tissue after 10 min of anoxia and the evoked response from the dentate granule cells again shows little recovery (10%). 4. The evoked response in the CA1 pyramidal cell layer of slices from young rats is more resistant to damage from 5 or 7 min anoxia than it is in slices from adults. Thus this region, also, shows an age-dependent increase in susceptibility to anoxic damage. ATP levels in the CA1 region of tissue from young animals at the end of 5 and 7 min anoxia are greater than ATP levels in tissue from adult animals after these same anoxic exposures. 5. Basal levels of 45Ca accumulation are greater in CA1 and dentate gyrus from young rats. However, the percentage increases during 10 min of anoxia are less than one-half the values in slices from adult animals. 6. The results suggest that the increased resistance of slices from young animals to anoxic transmission damage may be explained by the better maintenance of ATP in synaptic regions of these slices during anoxia. This may confer the increased resistance by lowering the anoxic increase in cell Ca2+. PMID- 2557435 TI - Spatial spread of in-field afferent inhibition in the cat's spinocervical tract. AB - 1. Extracellular microelectrode recordings were made from twenty-three spinocervical tract (SCT) cells in the lumbar spinal cord of cats anaesthetized with chloralose and paralysed with gallamine triethiodide. Excitation and inhibition of the cells were elicited by applying small brief (4 mN, 60 ms) localized jets of air to the clipped hair in and around the receptive fields. 2. Receptive field extents ranged from 40 to 180 mm. Excitation occurred in the period 30-130 ms after the start of the stimulus, and in-field afferent inhibition from 130 ms up to 700 ms or more. The inhibition was manifest as a reduction in background discharge and as a reduction in responsiveness to a test stimulus which followed a conditioning stimulus. 3. When the conditioning stimulus was spatially separated from the test stimulus, the degrees of in-field afferent inhibition depended on the spatial separation, even when both were within the excitatory receptive field. The spatial spread of in-field afferent inhibition was limited to 100 mm or less. 4. In two units only, afferent inhibition was produced from a narrow strip just outside the excitatory receptive field. In the other units, it could only be produced from within the excitatory receptive field. 5. The results suggest that the inhibitory input to SCT cells is organized in subdomains no more than 100 mm across, which may correspond to the receptive fields of interneurones between the primary afferent fibres and the SCT cells. PMID- 2557436 TI - Properties of calcium channels in guinea-pig gastric myocytes. AB - 1. The inward membrane current in enzymatically dispersed guinea-pig gastric myocytes was studied using whole-cell voltage clamp technique. 2. Only one inward membrane current was found in gastric myocytes which was identified as the Ca2+ current based on its inhibition by Ni2+, Cd2+ and Co2+, its dependence on [Ca2+]o, and its insensitivity to variations of [Na+]o. 3. Ca2+ current activated at -20 mV, peaked around +10 mV and was markedly enhanced when the holding potential was increased from -40 to -90 mV. The enhancement of ICa at negative holding potentials did not alter the activation threshold of ICa. When Ba2+ was substituted for Ca2+, IBa was similarly enhanced at more negative potentials. 4. In cells where internal Ca2+ was buffered with 10 mM-EGTA, the time course of inactivation was fitted with two exponentials, with time constants: tau f = 53.4 +/- 18.1 ms and tau s = 175.2 +/- 46.1 ms. When Ba2+ was the charge carrier through the channel, the time course of inactivation could be fitted often by only one exponential which approximated tau s for inactivation of ICa. The voltage dependence of steady-state inactivation of Ca2+ channels was not significantly altered when Ba2+ was the charge carrier. 5. Using different buffering systems (EGTA, EDTA and citrate), we found that citrate maintained the ICa and slowed inactivation more effectively than the other buffers tested. Because the calculated change in [Ca2+]i did not differ significantly between buffer systems, we speculate that suppression of inactivation by citrate is related to increased accessability of the buffer to cytoplasmic Ca2+ near the Ca2+ channel. Changes in [Mg2+]i affected peak ICa but not the kinetics of inactivation indicating that [Mg2+]i may regulate the steady-state inactivation or the availability of the Ca2+ channels. 6. The divalent selectivity of the Ca2+ channel had the following sequence: Ba2+ greater than Ca2+ greater than or equal to Sr2+ much greater than Mg2+. In very low extracellular Ca2+ (less than 10(-7) M), the Ca2+ channel conducted Na+. 7. Increasing [H+]o appeared to differentially affect peak and maintained components of ICa. At pH less than 6.5, the maintained component of ICa was suppressed more than the peak component indicating possible time- and voltage-dependent inhibition of ICa by protons. 8. Nifedipine, D600 and diltiazem inhibited ICa in a voltage-dependent manner. The order of potency for inhibition of peak ICa was nifedipine approximately D600 much greater than diltiazem.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557438 TI - A cation channel in the thick ascending limb of Henle's loop of the mouse kidney: inhibition by adenine nucleotides. AB - 1. Patch-clamp single-channel current recordings were used to study the inhibition of Ca2+-activated non-selective cation channels by internal nucleotides in patches excised from basolateral membranes of the thick ascending limb of Henle's loop of the mouse kidney. 2. The application of ATP, ADP or AMP to the cytoplasmic face of excised inside-out membrane patches reduced the open state probability of the channels (Po) in a dose-dependent way without effect upon the unitary current amplitude. Dose-response curves gave half-maximal inhibitory concentrations of 20, 21 and 2.5 microM for ATP, ADP and AMP, respectively, while the Hill coefficient was close to one in all three cases. 3. Cyclic AMP partially inhibited channel activity (Po = 35 +/- 17% of control) only at high, unphysiological concentrations (10(-3) M) while adenosine (10(-3) M) had very little effect (Po = 83 +/- 7% of control). 4. Replacement of adenine with other purines (guanine, hypoxanthine) or pyrimidine (uridine) bases very largely reduced inhibitory activity. Cyclic GMP had no effect. 5. Non-hydrolysable analogues of ATP, AMP-PNP (10(-3) M) and ATP-gamma-S (5 x 10(-4) M), were effective inhibitors of the channel (Po = 24 +/- 7 and 9 +/- 4% of control, respectively. PMID- 2557437 TI - Interaction between calcium channel ligands and guanine nucleotides in cultured rat sensory and sympathetic neurones. AB - 1. Voltage-activated Ca2+ channel currents were recorded from cultured rat dorsal root ganglion (DRG) neurones using the whole-cell clamp technique with Ba2+ as the charge carrier. 2. Inclusion of the GTP analogue guanosine 5'-O-3 thiotriphosphate (GTP-gamma-S, 500 microM) or guanylylimidodiphosphate (GMP-PNP, 500 microM) or GTP itself (1 mM) in the patch pipette solution resulted in a smaller, slowly activating Ca2+ channel current which did not inactivate during a 100 ms voltage step. This current was inhibited by CdCl2 (10-100 microM) and omega-conotoxin (1 microM). 3. Nifedipine (5 microM), (-)-(R)-201-791 (5 microM), D600 (10 microM), and diltiazem (30 microM) inhibited Ca2+ channel currents recorded from control neurones, although in some cells a biphasic response was observed, with an initial increase preceding the inhibition of the currents. In the presence of internal GTP-gamma-S, at a holding potential (VH) of -80 mV, only potentiation of the Ca2+ channel current was observed in the presence of all three Ca2+ channel ligands. Internal GMP-PNP, while less effective than GTP-gamma S, also resulted in D600 showing an agonist response. Similarly, in the presence of internal GTP (1 mM), (-)-(R)-202-791 gave a prolonged agonist response. 4. Nifedipine, whether acting as an antagonist in control cells or as an agonist in GTP-gamma-S-containing cells, induced a shift to more hyperpolarized potentials of the steady-state inactivation curves. 5. Potentiation of Ca2+ channel currents induced by D600 in GTP-gamma-S-containing cells, was not observed when the neurones were pre-treated with pertussis toxin. The presence of internal GDP-beta S (500 microM) did not significantly alter the maximum inhibitory action of D600 compared with controls. However, 1 mM-GDP-beta-S increased the rate of onset of inhibition by (-)-(R)-202-791. 6. Depolarizing VH to -30 mV accelerated the onset of inhibition induced by the Ca2+ channel ligands in control cells. In the presence of internal GTP-gamma-S at VH -30 mV, biphasic responses were produced by all the Ca2+ channel antagonist ligands with initial stimulation for 1-2 min being followed by inhibition of the Ca2+ channel currents. 7. The agonist actions of (+)-(S)-202-791 were potentiated by the presence of internal GTP-gamma-S. 8. The expression of an agonist response to (-)-(R)-202-791 induced by internal GTP gamma-S was also present in sympathetic neurones cultured from adult rat superior cervical ganglion (SCG).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557439 TI - Electrophysiological responses of dissociated type I cells of the rabbit carotid body to cyanide. AB - 1. The carotid body is the major peripheral sensor of arterial PO2 in the mammal and is excited by cyanide (CN-). Type I cells, the presumed sites for transduction, were freshly dissociated from the carotid body of the adult rabbit and studied with the whole-cell patch clamp technique. 2. Type I cells were hyperpolarized by CN-, the action potential was shortened, and there was an increased after-hyperpolarization. 3. Under voltage clamp control, CN- increased a voltage-dependent outward current, which showed pronounced outward rectification. Tail currents increased by CN- reversed close to the predicted EK, the reversal potential of the CN--induced current depended on extracellular [K+], and the current was blocked by intracellular TEA+ and Cs+. 4. The i-V relation of the CN--induced conductance strongly mirrored that of voltage-gated Ca2+ entry, and the response was abolished by removal of extracellular Ca2+. We conclude that the increased gK is Ca2+ -dependent (gK(Ca]. 5. The Ca2+ current was attenuated by CN-, and showed an increased rate of inactivation. Thus, the increased gK(Ca) must result from an alteration in Ca2+ homeostasis independent of the Ca2+ current, and not an increased Ca2+ entry through voltage-activated channels. 6. Carbachol also hyperpolarized cells and increased a K+ conductance. 7. At depolarized holding potentials a steady-state outward current was increased by CN . The current reversed close to EK, and was associated with increased current fluctuations. Noise analysis showed that a channel conductance of 3 pS carries the current. 8. The response to CN- was not impaired by the inclusion of 5 mM MgATP in the patch pipette. 9. If signals to the CNS are initiated by the calcium dependent release of transmitters from type I cells, transduction would appear to be the direct consequence of the energy dependence of Ca2+ homeostasis. PMID- 2557440 TI - The effect of the phenylalkylamine D888 (devapamil) on force and Ca2+ current in isolated frog skeletal muscle fibres. AB - 1. The effects of the (+)- and the (-)-isomer of the phenylalkylamine derivative D888 (desmethoxyverapamil or devapamil) on isometric force and slow Ca2+ inward current were investigated in short toe muscle fibres of the frog (Rana temporaria). The experiments were performed under voltage-clamp conditions with two flexible internal glass microelectrodes at 10 degrees C in a TEA sulphate solution containing approximately 4 mM-free Ca2+. 2. In the presence of 0.05-5 microM-(-)-D888 a normal phasic contracture could be induced by a depolarizing voltage step. When depolarization was maintained for some minutes the force controlling system turned into a stabilized inactivated state (paralysis) from which it recovered upon repolarization within minutes instead of seconds. With the (+)-isomer (0.5-20 microM), a similarly retarded restoration was observed. However, it proved to be less effective than the (-)-isomer. 3. D888 caused a shift to more negative potentials of the S-shaped curve, which describes the voltage dependence of force restoration in the steady state (restoration time 15 min). The potential of half-maximum restoration in the absence of the drug (V = 35.8 mV) changed as follows. (-)-D888: -56 mV (0.05 microM), -69 mV (0.2 microM), -77.5 mV (0.5 microM), and -82 mV (5 microM); (+)-D888: -55.8 mV (0.5 microM), 76.5 mV (5 microM), and -85 mV (20 microM). 4. On the assumption that D888 binds only to the inactivated form of the voltage sensor of force control in the T tubular membrane (modulated receptor hypothesis) the data presented in paragraph 3 allowed an estimation of the drug-receptor dissociation constants. The KD values ascertained in this way, 1.71 nM for the (-)-isomer and 12.9 nM for the (+)-isomer, are in fair agreement with those obtained from [3H]D888 binding studies by other authors. 5. A comparison between equal concentrations of the two isomers regarding their effect on the speed of restoration and the time needed to transform the sensor into the paralysed state suggests that the differences in the dissociation constants are mainly due to a greater dissociation rate of the (+)-isomer from the sensor. 6. The restoration of the Ca2+ channel was retarded by D888 to a similar extent as that of the voltage sensor. This parallel action on both systems indicates structural similarities between the voltage sensor and the Ca2+ channel. 7. It is concluded that D888 'stabilizes' the inactivated state of the voltage sensor and the Ca2+ channel in a way similar to D600, but with a higher potency. Both isomers of D888 showed an antagonistic action and differed only in their potency. PMID- 2557441 TI - A T-type Ca2+ current underlies low-threshold Ca2+ potentials in cells of the cat and rat lateral geniculate nucleus. AB - 1. The characteristics of a transient inward Ca2+ current (IT) underlying low threshold Ca2+ potentials were studied in projection cells of the cat and rat dorsal lateral geniculate nucleus (LGN) in vitro using the single-electrode voltage-clamp technique. 2. In cat LGN slices perfused at 25 degrees C with a solution which included 1 mM-Ca2+ and 3 mM-Mg2+, IT could be evoked by depolarizing voltage steps to -55 mV from a holding potential (Vh) of -95 mV and was abolished by reducing [Ca2+]o from 1 to 0.1 mM. IT was also blocked by 8 mM Mg2+ and 500 microM-Ni2+, but 500 microM-Cd2+ was a significantly less effective antagonist. 3. The inactivation of IT, which occurred at Vh positive to -65 mV, was removed as Vh approached -100 mV. The process of inactivation removal was also time dependent, with 800-1000 ms needed for total removal. Activation curves for IT showed a threshold of -70 mV and illustrated that IT was extremely voltage sensitive over the voltage range from -65 to -55 mV. 4. The decay phase of IT followed a single-exponential time course with a time constant of decay which was voltage sensitive and ranged from 20 to 100 ms. The mean peak conductance increase associated with IT was 8.4 nS (+/-0.9, S.E.M.). 5. In more 'physiological' conditions (35 degrees C and 1.5 mM-Ca2+, 1 mM-Mg2+) the voltage dependence of activation and inactivation were unaffected. However, the development and decay of IT proceeded more rapidly and only 500-600 ms were needed for total removal of inactivation. Under these conditions, the use of voltage ramps showed that depolarization rates of greater than 30 mV/s were necessary for IT activation. 6. The use of multiple voltage-step protocols illustrated that the process of inactivation removal was rapidly reversed by brief returns to a Vh of -50 mV. Furthermore, any delay in IT activation, once the LGN cell membrane potential was in the IT activation range, resulted in a current of reduced amplitude. 7. Although IT in rat LGN cells was briefer and had a shorter latency to peak, it was otherwise similar to that seen in cat LGN cells. 8. The characteristics of IT are very similar to those of the T-type Ca2+ currents of other excitable membranes. The properties of IT are discussed with respect to its role in generating the low-threshold Ca2+ potentials which are central to the oscillatory behaviour of thalamic projection cells. PMID- 2557442 TI - Characterization of proton currents in neurones of the snail, Lymnaea stagnalis. AB - 1. Internal perfusion voltage-clamp and inside-out patch-clamp techniques were used to study the voltage-dependent H+ currents in snail neurone cell bodies. 2. In whole cells the voltage-activated outward H+ current was measured 60 ms after stepping to +40 mV with an internal pH (pHi) of 5.9 and no internal K+([K+]i = 0), and the delayed K+ current was measured 60 ms after stepping to +40 mV with pHi = 7.3 and [K+]i = 74 mM. The mean H+ and K+ current densities were 14.6 +/- 7.8 and 38.2 +/- 14.0 nA/nF, respectively, giving a mean ratio of the H+ to K+ current of 0.4 +/- 0.2. There is not a strong correlation between the densities of the two kinds of outward currents found in different cells. 3. Inside-out patch studies reveal that the H+ and K+ currents are distributed quite differently in the membrane. While 85% of all patches had K+ current, only five out of thirty-eight patches studied had H+ currents. In those five patches the H+ currents measured at +30 mV ranged from 10.7 to 21.0 pA, and the ratio of the H+ and K+ currents at +30 mV was 0.83 +/- 0.38. The mean H+ and K+ currents for all thirty-eight patches were 1.9 +/- 4.9 and 10.5 +/- 7.9 pA, respectively. 4. The current distribution patterns demonstrate that the H+ current does not flow through the delayed K+ current channels even though the two currents have similar voltage dependence and time course. 5. The relative ability of various extracellular divalent cations to block the H+ current was found to be Cu2+ approximately equal to Zn2+ greater than Ni2+ greater than Cd2+ greater than Co2+ greater than Mn2+ greater than Mg2+ = Ca2+ = Ba2+. Since 100 microM-Zn2+ blocks the H+ current more than it blocks the Ca2+ current, it can be used to reduce the contamination of Ca2+ current measurements by the H+ current. 6. The magnitude of the H+ current has a stronger temperature sensitivity than does the magnitude of the delayed K+ current. The Q10 of the H+ current magnitude is 2.1 +/- 0.4, while the Q10 of the K+ current magnitude is 1.4 +/- 0.04. This suggests a higher activation energy may be involved in the conduction of the H+ current than for K+ current. 7. The smooth time course of the H+ current measured in patches indicates that the size of the unitary H+ current is very small.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557444 TI - Characterization of a cytochalasin D-resistant mutant of Entamoeba histolytica. AB - Characterization of a cytochalasin D-resistant mutant of the human parasite Entamoeba histolytica capable of growing at 10 microM cytochalasin is described. The mutant cells also show resistance to 5 mM colchicine and 100 microM cytochalasin B, drugs proved deleterious for wild type trophozoites. The mutants show increased osmotic fragility and electric mobility but reduced phagocytic activity, and agglutination by Concanavalin A. On the other hand pinocytic activity remains unaltered when compared with the wild type cells. Polymerized actin, seen by staining with phalloidin, often appears polarized to one end of the trophozoites and forms few of the endocytic invaginations found in wild type amebas. An altered distribution of part of the actin could explain the differences in surface properties and motility observed in the mutant amebas. PMID- 2557443 TI - Action of the major protease from Entamoeba histolytica on proteins of the extracellular matrix. AB - The action of the major protease from the parasitic protozoon Entamoeba histolytica, a cysteine protease of Mr 27,000-29,000, on some important proteins of the extracellular matrix has been studied. The isolated protease degraded the extracellular matrix proteins from human tissue collagen type IV and V as well as laminin and fibronectin with different velocities and specificities under native conditions. Whereas the degradation of fibronectin and laminin proceeded rapidly, yielding distinct fragment patterns, the breakdown of the collagen types happened more slowly and incompletely. The digestion of the denatured isolated alpha 2 chain of bovine collagen type I was very fast and unspecific requiring only 1/10 of the enzyme activities as compared with the other substrates mentioned above. Nearly 85% of the overall proteolytic activity of a soluble fraction of E. histolytica was strongly inhibited by antibodies against the purified histolytic protease as well as by cystatin from chicken egg white, a specific protein inhibitor of cysteine proteases. We conclude that the histolytic protease represents by far the highest portion of soluble proteolytic activity in E. histolytica which is sufficient to destroy the extracellular matrix of the host. PMID- 2557445 TI - Breakdown of glucopolysaccharides in Entamoeba histolytica by phosphorylase. AB - Homogenates of trophozoites of Entamoeba histolytica released glucose 1-phosphate from amylopectin, glycogen, and amylose in a ratio of 100:78:74 at glucopolysaccharide concentrations of 0.1%. By use of self-generating Percoll gradients this activity was shown to be particulate and associated with glycogen. The phosphorylase was extracted from the 40,000 g pellet in aqueous medium and purified to homogeneity by gel filtration on Fractogel TSK HW-55(F) followed by chromatography on Blue Sepharose CL-6B. The purified enzyme was active not only against the glucopolysaccharides but also on dextrins with more than 3 glucose moieties, which were primarily formed by the action of amoebic amylases. At substrate concentrations of 1 mM nonreducing ends of each glucan, the phosphorolysis rate of the branched polysaccharides was about 1.75 x 10(4) times higher than those of the maltodextrins. By means of HPLC the sequential degradation of 4-nitrophenyl-maltoheptaoside (G(7)-pNP) was studied. Native phosphorylase exhibited a relative molecular mass of M(r) = 200,000 by gel filtration and gel electrophoresis. The SDS electrophoresis, under reducing conditions, indicated that the native enzyme was a dimer. Optimal degradation of the polysaccharides and dextrins was achieved at pH values of 7.5 and 7.0 respectively. PMID- 2557446 TI - Multiphase skeletal scintigraphy in primary fibromyalgia syndrome: a blinded study. AB - Sixteen patients with primary fibromyalgia syndrome were assessed blindly by multiphase skeletal scintigraphy to detect possible subclinical synovitis and uptake abnormalities at tender point sites. Results were normal in 14 of the 16 patients. Mild and localized abnormalities were found in 2 patients and were possibly due to the trauma of overuse. Our multiphase skeletal scintigraphy study showed that synovitis, other evidence of arthritis, or abnormalities at tender point sites were not increased above normal expectation among our sample of patients studied with fibromyalgia. PMID- 2557447 TI - Chronic coxsackie B infection mimicking primary fibromyalgia. AB - We describe a patient with primary fibromyalgia according to Smythe's criteria who had evidence of chronic infection with Coxsackie virus over a 4-year period. We discuss the association of Coxsackie infection and myalgic syndromes and review the literature. PMID- 2557448 TI - Polyarticular Clostridium perfringens pyoarthritis. AB - A woman with rheumatoid arthritis presented with an abdominal mass and Clostridium perfringens septic arthritis in both wrists and shoulders. We believed this to be the first reported case of polyarticular septic arthritis due to Clostridium perfringens. Apatite, lipid, and cholesterol crystals in the shoulder joint effusions were a potential source of diagnostic confusion. PMID- 2557450 TI - Responses of gamma-aminobutyrate receptor from rat brain: similarity of different preparation methods; muscimol induced desensitization and chloride exchange. AB - Chloride-36 exchange into three different membrane vesicle preparations from rat brain homogenate was followed. The different preparations all contained the same sealed vesicular components characterized by their rates of chloride exchange. The GABA-mediated 36Cl- exchange in all the preparations occurred in two phases shown to be mediated by two distinguishable receptors present in the activity ratio of 5:1 as previously described (Cash, D.J., Subbarao, K. 1987. Biochemistry 26:7556, 7562). Reported differences do not result from differences in the membrane preparations used or from the use of a GABA-mimetic instead of GABA, but from experimental differences. The preparations compared were made with mild or vigorous homogenization and with different extents of purification from solutes or membrane components: (i) a synaptoneurosome preparation, (ii) a Ficoll gradient preparation, and (iii) a washed P2 preparation. In each preparation the same four populations of membrane vesicles were characterized by their 36Cl- influx rates: (i) a major population (40-50%) (tau 1/2 = 1.4 min), (ii) a slower exchanging major population (40-55%) (tau 1/2 = 24 min), (iii) a minor population (5-12%) containing active GABA receptor and having the GABA-independent permeability of the slower exchanging population, and (iv) a very small exchange (approximately 2%) (tau 1/2 approximately 0.2 sec). The GABA-independent 36Cl- exchange processes were kinetically first order. The relative quantities of the different vesicle populations varied slightly with the preparation and purification technique. The identity of these components, observed in the different preparations, was attributed to the vesicle formation being dependent on the morphology and properties of the membrane rather than the preparation method. The soluble brain extract was GABA-mimetic with the two observed receptors, causing channel opening and desensitization. But little washing of the membrane was required to observe the function of both receptors. Muscimol was GABA-mimetic with both receptors. With muscimol, channel opening occurred at 2.6 fold lower concentrations while desensitization was unaltered relative to GABA. This is additional evidence that these responses are mediated by different pairs of binding sites. The dependence of desensitization rate on muscimol concentration indicated that there are two binding sites mediating desensitization, as described with GABA. PMID- 2557449 TI - Phosphate transport processes in eukaryotic cells. PMID- 2557451 TI - Reconstituted voltage-sensitive sodium channels from eel electroplax: activation of permeability by quaternary lidocaine, N-bromoacetamide, and N bromosuccinimide. AB - We have investigated the ion permeability properties of sodium channels purified from eel electroplax and reconstituted into liposomes. Under the influence of a depolarizing diffusion potential, these channels appear capable of occasional spontaneous openings. Fluxes which result from these openings are sodium selective and blocked (from opposite sides of the membrane) by tetrodotoxin (TTX) and moderate concentrations of the lidocaine analogue QX-314. Low concentrations of QX-314 paradoxically enhance this channel-mediated flux. N-bromoacetamide (NBA) and N-bromosuccinimide (NBS), reagents which remove inactivation gating in physiological preparations, transiently stimulate the sodium permeability of inside-out facing channels to high levels. The rise and subsequent fall of permeability appear to result from consecutive covalent modifications of the protein. Titration of the protein with the more reactive NBS can be used to produce stable, chronically active forms of the protein. Low concentrations of QX 314 produce a net facilitation of channel activation by NBA, while higher concentrations produce block of conductance. This suggests that rates of modifications by NBA which lead to the activation of permeability are influenced by conformational changes induced by QX-314 binding. PMID- 2557453 TI - Locations of nucleosomes on the regulatory region of simian virus 40 chromatin. AB - We have asked where the nucleosomes are located with respect to the replication origin and regulatory region of simian virus 40 DNA, what would be the possible functional consequences of the identified locations, and to what extent these locations correlate with the current views on mechanisms involved in establishing nucleosome-free regions in chromatin. To identify the precise location of nucleosomes, we have shot-gun cloned and sequenced nucleosomal DNA obtained from micrococcal nuclease digestion of wt776 chromatin prepared late in infection. Our results indicate that nucleosomes do not occupy unique positions over the replication origin or the elements involved in transcriptional control. However, it appears that the nucleosome distribution is not random, since several nucleosomes are represented by two or more independently generated clones. Two nearly identical cloned fragments map over the replication origin; five include 1.5 copies of the 72 base-pair enhancer sequences; and eight map to a region that spans a DNA bending locus and the major transcription initiation site of the late genes. The complex nucleosome distribution pattern observed in our direct analysis suggests that disparate nucleosome-free regions may be involved in controlling replication, and selective expression of the viral early or late genes. PMID- 2557452 TI - Mechanism of aldosterone-induced increase of K+ conductance in early distal renal tubule cells of the frog. AB - Isolated early distal tubule cells (EDC) of frog kidney were incubated for 20-28 hr in the presence of aldosterone and then whole-cell K+ currents were measured at constant intracellular pH by the whole-cell voltage-clamp technique. Aldosterone increased barium-inhibitable whole-cell K+ conductance (gK+) threefold. This effect was reduced by amiloride and totally abolished by ouabain. However, aldosterone could still raise gK- in ouabain-treated cells in the presence of furosemide. We tested whether changes in intracellular pH (pHi) could be a signal for cells to regulate gK+. After removal of aldosterone, the increase in gK+ was preserved by subsequent incubation for 8 hr at pH 7.6 but abolished at pH 6.6. In the complete absence of aldosterone, incubation of cells at pH 8.0 for 20-28 hr raised pHi and doubled gK+. Using the patch-clamp technique, three types of K+-selective channels were identified, which had conductances of 24, 45 and 59 pS. Aldosterone had no effect on the conductance or open probability (Po) of any of the three types of channels. However, the incidence of observing type II channels was increased from 4 to 22%. Type II channels were also found to be pH sensitive, Po was increased by raising pH. These results indicate that prolonged aldosterone treatment raises pHi and increases gK+ by promoting insertion of K+ channels into the cell membrane. Channel insertion is itself triggered by raising both pHi and increasing the activity of the Na+/K+ pump in early distal cells of frog kidney. PMID- 2557454 TI - Genetic analysis of the switch that controls porin gene expression in Escherichia coli K-12. AB - The two-component regulatory system, OmpR and EnvZ, in Escherichia coli controls the differential expression of ompF and ompC in response to medium osmolarity. Previous studies suggest that EnvZ functions as a membrane sensor relaying information to the DNA-binding protein, OmpR, which in turn activates expression of the appropriate promoter. A strategy has been devised to isolate and characterize a collection of missense mutations in ompR that alter, but do not abolish protein function. Mutants were isolated using strains that contain the ompR and envZ genes in separate chromosomal locations yet maintain the production of both regulatory proteins at physiological levels. Such an arrangement facilitates ompR diploid analysis and tests of epistasis with known envZ mutations. The data obtained indicate that OmpR works in both a positive and negative fashion to control the transcription of ompF and this result forms the basis of a model for porin regulation that explains the switch from OmpF to OmpC production in response to increasing medium osmolarity. PMID- 2557456 TI - Glucocorticoids modulate rat hypothalamic corticotrophin-releasing factor release induced by interleukin-1. AB - Continuous in vitro perifusion of rat hypothalami with interleukin-1 beta (IL-1) increased corticotropin-releasing factor (CRF) secretion in a dose-dependent manner, in the range of 1 to 5 U/ml. The stimulatory action of IL-1 was significantly attenuated by dexamethasone, both by addition to the perifusion medium and by in vivo dexamethasone pretreatment. The data suggest that an immunoneuroendocrine interaction circuit, subjected to glucocorticoid negative feedback, has an essential role in maintaining organism homeostasis. PMID- 2557455 TI - A polypeptide chain-refolding event occurs in the Gly82 variant of yeast iso-1 cytochrome c. AB - The replacement of Phe82 in yeast iso-1-cytochrome c by a glycine residue substantially alters both the tertiary structure and electron transfer properties of this protein. The largest structural change involves a polypeptide chain refolding of residues 79 through 85. Refolding places glycines 82, 83 and 84 immediately adjacent to the plane of the heme group in a spatial positioning comparable to that of the phenyl ring of Phe82 in the wild-type protein. Despite this perturbation in structure, solvent accessibility computations show that heme solvent exposure has not increased in the Gly82 variant protein. However, refolding does result in the introduction of a number of polar groups into the hydrophobic heme pocket. This appears to be responsible for the decreased reduction potential of the heme in this protein. The present study, along with that of the Ser82 variant protein (Louie et al., 1988b), clearly establishes the link between dielectric constant within the heme crevice and reduction potential. The further anomalously low electron transfer activity of the Gly82 variant protein would appear to arise from two factors. First, the polypeptide chain medium now adjacent to the heme is unable to facilitate electron transfer in a manner similar to that of the aromatic side-chain of Phe82. Second, polypeptide chain refolding significantly alters the surface contour of the Gly82 protein rendering it less suitable to interact with the corresponding complementary surfaces of redox partners. Our data support the conclusion that Phe82 plays a number of roles in the electron transfer process mediated by yeast iso-1 cytochrome c. These include the maintenance of the heme environment, provision of an optimal medium along the path of electron transfer and formation of interactions at the contact interface in complexes with redox partners. PMID- 2557457 TI - Glial cells of the oligodendrocyte lineage express proton-activated Na+ channels. AB - Neurons and oligodendrocytes, but not type I astrocytes and Schwann cells, generate large Na+ currents in response to a step increase of [H+]. Proton activated Na+ channels are the first cationic channels expressed in neuronal precursor cells from the mammalian brain. Glial precursor cells cultured from mouse brain are also capable of generating Na+ currents in response to step acidification (INa(H]. With further development along the oligodendrocyte lineage, this property is retained, whereas voltage-activated Na+ and K+ currents disappear. Comparing INa(H) of oligodendrocytes with INa(H) of their precursor cells did not reveal a difference in current amplitude, suggesting a higher density of INa(H) channels on the (smaller) precursor cells. The properties of INa(H) in glial precursor cells and oligodendrocytes are similar to those of neurons, with respect to activation conditions, time course, and the effect of extracellular Ca2+ concentrations. The results are consistent with previous observations which showed that oligodendrocytes partially preserve their chemically activated, but completely lose their voltage-activated, ion channels. PMID- 2557458 TI - DNA probe technique for diagnosis of human cytomegalovirus infection. AB - A rapid diagnostic assay for human cytomegalovirus (HCMV) has been developed for detecting HCMV DNA in urine samples with 32P-labelled cloned fragment, Eco RI fragment B, of DNA from HCMV strain Towne. 3.2 pg of homologous fragment from HCMV DNA could be detected by the labelled probe, and it did not hybridize DNA from other herpes viruses or human cells in dot hybridization assay. The assay correctly identified all (100%) of 7 coded urine specimen cultures positive for HCMV and 9 (90%) of 10 urine sample cultures negative for HCMV. So the hybridization assay was correct and as sensitive as the currently available tissue culture technique. The infection levels of different populations, such as organ transplantation recipients, patients with infantile hepatitis syndrome, normal infants, fetuses, have been investigated by the hybridization assay in the present study. PMID- 2557459 TI - Oxygen-derived free radicals related injury in the heart during ischemia and reperfusion. AB - It has been suggested recently that oxygen-derived free radicals may play an important role in the genesis of reperfusion injury and arrhythmias. Free radicals have a very short half-life (ranging from mili- to microseconds), hence almost all the reports supporting the free radical hypothesis of reperfusion cell injury have been indirect. We have applied electrone spin resonance spectrometry to measure directly the amount of free radicals generated during ischemia and reperfusion. The concentration of free radicals in mitochondria increased significantly during ischemia (for 20 and 40 min). The concentration of free radicals after reperfusion was higher than that during ischemia, and a large amount of free radical generation occurred within the first 60 sec of reperfusion and returned to the level of prereperfusion at 5 min after reperfusion. The concentration of free radicals in the reperfusion-induced ventricular fibrillation group was significantly higher than that in the non-occurrence group. The administration of liposomal superoxide dismutase reduced the incidence of reperfusion-induced ventricular fibrillation and that prevented the free radical generation during reperfusion. This study showed that enhanced generation of free radicals occurred at the onset of ventricular fibrillation and that free radical scavenger prevented the development of arrhythmias and free radical generation during reperfusion. We have obtained more circumstantial evidence for an involvement of free radicals in the genesis of reperfusion injury and arrhythmias. PMID- 2557460 TI - Possible mechanism responsible for mechanical dysfunction of ischemic myocardium: a role of oxygen free radicals. AB - It has been proposed that a major target organelles damaged by the ischemic process, probably by the oxygen free radicals generated, is the portion of the excitation-contraction coupling system that regulates Ca2+ delivery (the sarcoplasmic reticulum and sarcolemma) to the contractile proteins. We tested this hypothesis by studying the effect of in vitro generation of oxygen free radicals from xanthine-xanthine oxidase system or dihydroxyfumarate (DHF)/Fe3+ ADP system on Ca2+ flux behavior of canine cardiac sarcoplasmic reticulum (SR); sarcolemmal (Na+, K+)-ATPase and Na+-Ca2+ exchange activities; and myofibrillar (Ca2+, Mg2+)-ATPase activity. Generation of oxygen free radicals by xanthine oxidase acting on xanthine as a substrate increased the passive Ca2+ efflux and decreased intravesicular Ca2+ with no effect on active Ca2+ influx (Ca2+-ATPase) of SR vesicles. Similar exposure of sarcolemmal vesicles to xanthine plus xanthine oxidase stimulated Na+-Ca2+ exchange activity. When sarcolemmal vesicles were incubated with DHF plus Fe3+-ADP, (Na+, K+)-ATPase activity was decreased. It is postulated that the SR Ca2+ efflux pathways but not catalytic activity of the Ca2+ pump and sarcolemmal (Na+, K+)-ATPase involving Na+-Ca2+ exchange activity are altered by oxygen free radicals, and such changes may partly account for the occurrence of intracellular Ca2+ overload during the course of myocardial ischemia. Interestingly, oxygen free radicals from xanthine-xanthine oxidase system had no effect on myofibrillar pCa-ATPase curve. From this set of observations we would hypothesize that the SR and sarcolemma may be the principal target organelles of oxygen free radicals attack in the ischemic injury and not the contractile proteins per se. PMID- 2557461 TI - [Lung cancer in chromium-exposed workers--high prevalence of lung cancer and multiple primary cancers]. PMID- 2557462 TI - [Synthetic female hormones-induced liver cancer]. PMID- 2557463 TI - [Progress in the diagnostic imaging of liver cancer]. PMID- 2557464 TI - [Surgical technics to increase resectional rate in liver cancer]. PMID- 2557466 TI - [Various factors related to carbohydrate metabolism. Biogenic amines]. PMID- 2557467 TI - [Detection of papilloma virus in the cervix of the human uterus by in situ hybridization--Comparative study of detection rate by Southern blot method]. AB - Cases found to be positive for human papilloma virus (HPV) infection in the cervix by Southern blot method were evaluated with a newly developed kit for in situ HPV tissue hybridization. By the in situ hybridization method, HPV-DNA was detected without damaging the tissue structure. It appeared as purple or black stains in the nucleus of the epithelial cells that were located at the level of one-third of the epithelial thickness from the surface. The cases that were negative with the Southern blot method were also negative with the in situ hybridization method, but only 55.8% of the cases positive with the Southern blot method were positive with the in situ hybridization. Although in situ hybridization method is not as sensitive as the Southern blot method, it allows analysis of old paraffin blocks and comparison with pathological features. PMID- 2557465 TI - [Secondary leukemia (AML) with TcR beta rearrangement]. AB - A 60-year-old female with fever and general lymphadenopathy was admitted to our hospital in 1979. By histological examination of cervical lymph node, she was diagnosed as B-cell malignant lymphoma, diffuse, large, stage IV B (Ann Arbor classification). In 1984, the complete remission of malignant lymphoma was observed after MOPP, COPP and VAPP chemotherapies. After 3 years, she was readmitted because of leukocytosis in which 64% immature abnormal cells were detected. By conventional cytological and cytochemical evaluations, the immature abnormal cells were identified as secondary AML cells with M 2 type (FAB), peroxidase (+), My 7 (+) and chromosomal abnormality; 48, X, -X, +4, -9, +21, del (1) (p 34), i (17 q), + M 1, + M 2. Analysis of Ig and TcR genes showed TcR-beta rearrangement. It has been reported that secondary leukemia frequently causes chromosomal abnormalities and therapy resistance. Ig gene and TcR gene analysis of secondary leukemia combined with chromosome examination, which has reported little, may be useful in predicting prognosis and therapy resistance. PMID- 2557469 TI - [Destruction of intraportal tumor thrombus of hepatocellular carcinoma by laser irradiation]. PMID- 2557468 TI - [Detection of HPV-DNA in the various uterocervical lesions by in situ hybridization]. AB - We detected HPV (Human Papilloma Virus) -DNA in various uterocervical lesions by in situ hybridization using biotinylated DNA probes. In cases positive for HPV, the nuclei of the epithelial cells was purple to blackish purple. In 2 of 6 cases of chronic cervicitis, HPV-DNA was detected in the outer layer of the squamous epithelium. Eleven of 19 with mild dysplasia (57.9%) showed a positive reaction in the upper one-third of the epidermis in a mainly consisting of koilocytotic cells. All 6 patients with moderate dysplasia had positive cells among the koilocytotic cells and atypical cells in the middle layer. Five of 11 patients with severe dysplasia had scattered positive cells. Two of them had atypical condylomatoid lesions. Eight of 32 patients with squamous cell carcinoma were positive for HPV-DNA, but there was no consistent distribution pattern of the positive cells. PMID- 2557470 TI - [A case of cholangiocarcinoma who showed mimicking appearance of primary sclerosing cholangitis on the cholangiography]. PMID- 2557471 TI - [Two cases of digestive cancer demonstrated remarkable regression by hyperthermia with systemic chemotherapy]. PMID- 2557472 TI - Separation of human neutrophils in self-generated continuous density gradients of Percoll. AB - Human neutrophils were separated into two fractions using a continuous density gradient of Percoll solution. A marked decrease in O2- production was observed in the low-density neutrophil fraction. There was little erythrocyte contamination in the high-density neutrophil fraction (less than 0.5%), and thus the hypotonic or ammonium chloride lysis of erythrocytes was not necessary. PMID- 2557473 TI - [Effects of vasoactive drugs on isolated rabbit corpus cavernosum penis]. AB - Recently, intracavernous injection of some vasoactive drugs has been performed for the diagnosis and treatment of impotence. Despite extensive studies the mechanism of erection is still obscure. Therefore, the author studied the effects of some vasoactive drugs on isolated rabbit corpus cavernosum penis. Norepinephrine, phenylephrine or clonidine caused contraction of isolated rabbit corpus cavernosum strips in a concentration-dependent manner. This contractile effect was more potent with norepinephrine and phenylephrine than with clonidine. Prazosin was more effective than yohimbine in inhibiting norepinephrine-induced contractions. Papaverine and verapamil strongly relaxed the strips contracted by norepinephrine. Prostaglandin E1 also showed a relaxant effect. Low concentrations of isoproterenol caused relaxation, but in high concentrations it caused contraction. Acetylcholine relaxed norepinephrine-contracted strips in a concentration-dependent manner. Although the relaxant effect of acetylcholine was weaker than that of papaverine at high concentrations, acetylcholine and papaverine were almost equally effective at low concentrations. Vasoactive intestinal polypeptide relaxed the strips, and it was significantly more potent than papaverine. These findings suggest that both postsynaptic alpha 1 adrenoceptors and alpha 2-adrenoceptors are present in rabbit corpus cavernosum penis, and that alpha 1-adrenoceptors are predominant. The flaccid state of the rabbit penis seems to be maintained mainly by alpha 1-adrenoceptors. Verapamil seems to be useful for the diagnosis and treatment of impotence as papaverine. Acetylcholine and vasoactive intestinal polypeptide may be neurotransmitters involved in erection. PMID- 2557475 TI - Methemoglobin formation and reduction in canine erythrocytes with inherited high Na,K-ATPase activity. AB - Methemoglobin formation and reduction in canine erythrocytes with inherited high Na,K-ATPase activity (HK cells) were compared with those in normal canine cells (LK cells). Nitrite-induced methemoglobin formation in hemoglobin solutions indicated that the hemoglobin from HK cells was oxidized at essentially the same rate as that of LK cells. However, methemoglobin formation in HK cells was slower due to the inhibition by high glutathione (GSH) concentration. Methemoglobin reduction was allowed to take place on nitrite-treated and washed erythrocytes in a glucose medium and was reduced more rapidly in HK cells than in LK cells. During the reduction, the amounts of lactate and pyruvate increased more rapidly in HK cells, indicating enhanced glycolysis in HK cells. It is thus evident that the hemoglobin of HK cells is more securely protected from nitrite-induced oxidation by the GSH presence in great excess and by the increase in glycolysis. PMID- 2557474 TI - A comparison of polypeptides and restriction endonuclease sites of BHV-1 isolates and identification of IPV virus in Japan. AB - Bovine herpesvirus 1 (BHV-1) isolates from respiratory tract and from vagina of bovine in Japan were analyzed by SDS-polyacrylamide gel electrophoresis (SDS PAGE) and the DNA restriction endonuclease cleavage pattern, and compared with European BHV-1 strains. Both protein profile and DNA cleavaged pattern of BHV-1 isolates from respiratory tract were the same as those of European infectious bovine rhinotracheitis (IBR) virus, whereas the protein profile and DNA cleavage patterns of one isolate (M1) from vagina was the same as those of the European infectious pustular vulvovaginitis (IPV) virus. The facts indicate that IPV virus has existed in Japan. PMID- 2557476 TI - Glycosaminoglycans in transplantable rat malignant fibrous histiocytoma. PMID- 2557477 TI - Complement activation by infectious bursal disease virus and its relevance to virus growth in the lymphoid cells. PMID- 2557478 TI - Large and small plaque variants of canine adenovirus 2 isolate. PMID- 2557479 TI - An improved hemagglutination-inhibition test for pseudorabies virus. PMID- 2557480 TI - Platelet-derived growth factor receptors in the kidney--upregulated expression in inflammation. AB - Major features of a long-standing inflammation in the kidney are vascular proliferation, glomerulosclerosis, interstitial fibrosis and tubular atrophy, leading to a gradual deterioration of the renal function. In this study we have investigated the expression of B-type receptors for platelet-derived growth factor (PDGF) in frozen sections from normal and inflamed kidneys. Immunohistochemical techniques, employing two monoclonal antibodies specific for PDGF B-type receptors, were used. The specimens investigated were 15 kidneys removed by transplantectomy because of chronic rejection, 20 cases of glomerulonephritis with crescent formation, mesangial proliferation or non proliferative glomerulonephritis, and six normal kidneys. In parallel we characterized cellular infiltrates and class II transplantation antigen expression in the inflamed kidneys. An enhanced PDGF receptor expression was found on intimal cells and on smooth muscle cells of the proliferating vessels, on glomerular cells in glomeruli with mesangial proliferation, and on fibroblast like cells in the proximity of clusters of infiltrating macrophages and T lymphocytes of the interstitial tissue. Induction of PDGF receptor expression may render cells responsive to stimulation by PDGF, released from PDGF-producing cells, such as activated macrophages and from platelets. Our data suggest that PDGF is involved in the proliferation of mesenchymal cells that is seen in rejected kidney transplants and glomerulonephritis. PMID- 2557481 TI - Optimal correction of acidosis changes progression of dialysis osteodystrophy. AB - To investigate an eventual role of acidosis on hemodialysis osteodystrophy we prospectively studied 21 patients who were dialyzed with different amounts of bicarbonate in the dialysate for 18 months. According to the level of bone formation rate (BFR) on a prestudy bone biopsy, patients were split in two subgroups. Inside these two subgroups patients were randomly allocated to two therapeutics groups: 10 patients (group A) were dialyzed with the conventional amount of bicarbonate (33 +/- 2 mmol/liter) in the dialysate; the rest of the patients (group B, N = 11) had 7 to 15 mmol/liter sodium bicarbonate added to the dialysate to obtain 24 mEq predialysis bicarbonate plasma levels. An effective correction of acidosis was shown in group B by a higher predialysis plasma bicarbonate level (15.6 +/- 1 group A vs. 24.0 +/- 0.6 mEq/liter group B, P less than 0.005), which was reached three months after start of the study. Compared to the prestudy bone biopsy, osteoid and osteoblastic surfaces increased in group A but not in group B on the bone biopsies performed at the end of the study. Parathormone plasma level (iPTH), measured with an antiserum which cross reacts with the 44-68 region of PTH molecule, increased during the study in group A but not in group B. This finding suggested progression of secondary hyperparathyroidism (HPT) only in group A patients. Osteocalcin plasma values increased in both groups during the 18 months of the study. Consequently the two subgroups of patients formed on the basis of BFR level were evaluated separately.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557482 TI - In vitro modulation of tubular cyst regression in murine polycystic kidney disease. AB - Recent studies in a murine model of genetically-determined polycystic kidney disease, the CPK mouse, have suggested that alterations in renal Na-K ATPase activity in concert with tubular epithelial hyperplasia have pathogenic import in proximal tubular cyst formation. In the current study, we therefore studied the relative roles of Na-K ATPase activity, tubular epithelial hyperplasia, and basal lamina alterations during in vitro modulation of proximal tubular cyst regression during serum-free organ culture of newborn CPK kidneys. Under basal in vitro conditions, regression of CPK proximal tubular cysts was demonstrated in association with progressive decreases in Na-K ATPase activity and tubular epithelial hyperplasia. The pattern of proximal tubular cyst regression was modified by: a) Na-K ATPase activity induction with triiodothyronine, which promoted proximal tubular cystogenesis; and b) Na-K ATPase activity inhibition with ouabain, which blocked the effects of T3 on the process of cyst formation. Modulation of proximal tubular cystogenesis by Na-K ATPase induction and inhibition were accomplished without significant changes in proximal tubular epithelial hyperplasia or expression of basal lamina components. We conclude that increased Na pump activity may have a significant role in proximal tubular cyst formation and progressive enlargement in the CPK mouse. PMID- 2557483 TI - Long-term enalapril and verapamil in rats with reduced renal mass. AB - The effect of long-term treatment with either enalapril or high dose verapamil on survival, proteinuria, blood pressure and renal morphology was studied in female Wistar rats with markedly reduced renal mass. Four weeks were allowed for remnant kidney hypertrophy before determining the response to renal ablation of individual animals regarding proteinuria and hypertension. At this time, five groups of 18 rats were formed with equal levels of proteinuria and hypertension. Groups E1 and E2 were treated with enalapril, groups V1 and V2 with verapamil, and one group served as control. The daily food allowance was 14 g/rat of a standard rat diet, containing 30% protein and 100 mmol NaCl/kg food in groups E1 and V1. NaCl content was reduced to 20 mmol/kg food in groups E2, V2 and control. The drugs were added to the drinking water, enalapril at a dose of 0.1 g/liter, verapamil at 0.5 to 0.7 g/liter. Drug intake thus amounted to 10 to 25 mg/kg for enalapril and 50 to 140 mg/kg for verapamil. Treatment was continued for 15 weeks. Three of the 18 control rats did not survive up to 15 weeks. Mortality was lower in the enalapril treated groups with a single nonsurvivor in group E1. In contrast, mortality was higher in the verapamil treated animals with seven nonsurvivors in group V1 and eight in group V2. Blood pressure control was excellent in both enalapril treated groups. and proteinuria decreased in most animals of group E1 and all of group 22. Glomerulosclerosis did not develop in the majority of the enalapril treated animals. Despite the high dose, verapamil barely lowered blood pressure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557484 TI - Glomerular leukotriene synthesis in Heymann nephritis. AB - The glomerular synthesis of LTB4 was assessed in glomeruli isolated from rats with passive Heymann nephritis (PHN). PHN was induced by a single intravenous administration of proteinuric doses of immune sera raised in sheep against rat brush border tubular fraction Fx1A. At various time points following induction of the disease glomeruli were isolated and LTB4 synthesis was assessed under basal and phospholipase A2 activation conditions. LTB4 was measured by high pressure liquid chromatography and radioimmunoassay and was identified by UV spectroscopy. The role of complement system in mediating glomerular LTB4 synthesis was also assessed in a group of decomplemented rats using cobra venom factor and at various time points following administration of immune serum. Following induction of PHN, enhanced glomerular LTB4 synthesis was observed as early as one hour, peaked at five hours and returned toward control levels over the subsequent four days. The peak in glomerular LTB4 synthesis did not correlate with changes in glomerular neutrophiles or macrophages. A second increment of LTB4 synthesis occurred at the onset of heavy proteinuria (day 5). Complement depletion reduced proteinuria and the enhancement in LTB4 synthesis at day 5 but had no effect at earlier time points. The observations indicate that in non-inflammatory forms of glomerular immune injury the glomerular arachidonate 5-lipoxygenation is enhanced. This phenomenon has no apparent relationship with increased glomerular permeability to protein and may reflect the presence or activation of a leukotriene producing cell following intraglomerular interactions of Fx1A antigen, anti-Fx1A antibody and complement. PMID- 2557485 TI - [Effect of light on the eye on metabolism and hormones]. AB - Numerous metabolic parameters in serum and urine were examined in 110 cataract patients before and after cataract surgery. The marked reduction in light passing through the eye due to opacities (vision less than 1/10) leads to characteristic metabolic and hormonal disturbances. ACTH and cortisol production decreases, metabolism slows down and due to an adrenal insufficiency for which the pituitary is responsible there are characteristic changes in the cortisol-dependent metabolic processes. In addition, an "energetic action" of the light affecting the hypothalamus via the retino-hypothalamic pathways (the "energetic portion" of the visual pathway) was proved in patients who were blinded by cataract and had metabolic disturbances as a result. Postoperatively, after elimination of the lens opacities, the metabolism and hormones of the same patients returned to normal. As a result of restoration of exogenous light stimulation to the diencephalon-hypophysis system via the retinohypothalamic pathway ("energetic pathway" of the optic system) the metabolism and hormones returned to normal during the patients' stay in the hospital. These comparative investigations in the same patients before and after cataract extraction provide for the first time irrefutable scientific evidence of the influence of light via the eye on the human organism. PMID- 2557486 TI - [Apparent central facial paralysis in Lyme borreliosis]. AB - In childhood Lyme-Borreliosis often presents as facial palsy. This progredient infectious disease is to be treated by intravenous penicillin therapy in order to avoid future complications. In three out of four own observations facial palsy raised suspicion towards central origin. By performance of the orbicularis oculi reflex in all four cases peripheral damage of the facial nerve could be demonstrated. This simple electromyographic method seems to be a valuable tool in the differentiation of central and peripheral facial nerve palsy in childhood, preserving the affected children from painful electrodiagnostic procedure or useless search for supranuclear lesions. PMID- 2557487 TI - Molecular markers of primitive neuroectodermal tumors and other pediatric central nervous system tumors. Monoclonal antibodies to neuronal and glial antigens distinguish subsets of primitive neuroectodermal tumors. AB - Seventy-one tumors of the central nervous system in children were studied immunohistologically. Thirty-seven were classified histologically as PNETs, of which 35 were located in the cerebellum (medulloblastomas), one in the cerebrum, and one in the spinal cord. The 34 non-PNETs included five ependymomas, seven gangliogliomas, 15 astrocytomas, and seven tumors of other histology. We used monoclonal antibodies specific for neurofilament (NF) triplet proteins, for microtubule associated protein 2 and tau protein and for glial fibrillary acidic protein (GFAP) and myelin basic protein. In addition, a monoclonal antibody to epithelial membrane antigen was applied. The presence or absence of these antigens defined four major groups of PNETs: 1) PNETs not otherwise specified (10 cases), 2) PNETs with neuronal differentiation (eight cases), 3) PNETs with astrocytic differentiation (six cases), and 4) PNETs with both neuronal and astrocytic differentiation (12 cases). One case showed ependymal differentiation. The pattern of expression of NF isoforms in PNETs was reminiscent of that seen during normal mammalian development, such that phosphorylated NF-H was only present in combination with NF-M and NF-L. Among the other central nervous system tumors, all astrocytomas and gangliogliomas were positive for GFAP, and the gangliogliomas also expressed all NF isoforms. Three atypical teratoid tumors and two rhabdoid tumors showed strong positivity for epithelial membrane antigen and also for GFAP. We conclude that the differentiation antigens described here serve to distinguish PNETs from other pediatric central nervous system tumors and to identify subsets of PNETs. Accordingly, PNETs represent a heterogeneous group of pediatric brain tumors capable of neuronal and glial differentiation. PMID- 2557488 TI - Viremia and glial nodule encephalitis after experimental systemic cytomegalovirus infection. AB - Despite the importance of cytomegalovirus (CMV) infection of the central nervous system in acquired immune deficiency syndrome, a well characterized laboratory model of glial nodule encephalitis after systemic CMV infection is not available. We now report that after intraperitoneal infection of young guinea pigs with CMV, infection of the brain was routinely found in the 2nd week. Recovery of virus from the brain was achieved at the time of viremia. Histopathologic changes in the brain followed the recovery of virus and continued beyond the point that virus was cleared from the brain. Microglial nodules, which were sometimes observed in association with intranuclear inclusion bearing cells, were the predominant feature. Other histopathology included perivascular infiltrates, vascular endothelial swelling, subependymal infiltrates, and sporadic focal leptomeningitis. In comparison to our previous studies after intracerebral inoculation, parenchymal changes dominated and leptomeningitis was found infrequently. The present studies suggest that focal central nervous system infection by CMV may be relatively common, though clinically silent, in the course of systemic infection. Relevant to acquired immune deficiency syndrome, the model should facilitate studies of the mechanism of brain infection, local central nervous system host defense, and mechanisms of injury to the brain. PMID- 2557489 TI - Modulation by sex steroids of brain opioid receptors: implications for the control of gonadotropins and prolactin secretion. AB - Several experiments have been performed in order to analyze whether physiological or experimental changes of the endocrine environment might modify the binding characteristics of brain mu and kappa opioid receptors in the brain of the female and male rat. (a) In a first series of experiments, it has been observed that in the whole brain of regularly cycling female rats the number of mu receptors shows variation during the different phases of the estrous cycle. In particular a significant increase of the number of mu receptors has been observed in the morning of proestrus and in the afternoon of estrus. (b) In a second series of experiments, it has been shown that the administration of estrogens brings about a significant increase in the number of mu receptors in the hippocampus and in the thalamus of ovariectomized rats, while the administration of a regime including estrogen and progesterone induces a significant decrease of the number of mu receptors in the hypothalamus and in the corpus striatum. These data seem to indicate that hypothalamic mu receptors may be involved in the positive but not in the negative feedback control of LH secretion. (c) In a third series of experiments, it has been found that the number of mu receptors in the whole brain of 15- and 22-month-old male rats and in the hypothalamus of 22-month-old male rats is significantly lower than in the same tissues of young animals; moreover, the administration to old animals of testosterone does not modify the number of hypothalamic mu opioid receptors, indicating that the decline of brain mu receptors in old animals is not the consequence of the physiological decline of testosterone secretion but probably represents an autonomous phenomenon. (d) In a fourth series of experiments, it was shown that, in young male rats, the concentration of kappa receptors is extremely variable in different regions of the brain. The highest concentrations have been found in the hypothalamus and in the striatum; also in the mesencephalon and in the amygdala kappa receptors are present in rather elevated quantities; lower concentrations have been found in the thalamus, the frontal poles, the hippocampus and in the anterior and posterior cerebral cortex. These experiments have shown in addition that the process of aging induces an increase of the number of kappa receptors in the amygdala and in the thalamus; no age-linked modifications were observed in the other structures examined.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557491 TI - [Current treatment of malignant ovarian germ cell tumors]. AB - Thirty-two patients were treated for ovarian germ cell malignomas, first, along with surgery, with a combined chemotherapy (vincristine, actinomycin D, cyclophosphamide) up to 18 treatments and later up to 6 treatments, except in immature teratomas and advanced cases. At present, in the third stage of the disease a new protocol is applied: cis-platinum, vinblastine, and bleomycin. Four patients with dysgerminoma were surgically treated (two were subsequently irradiated). Only two patients died--one with an endodermal sinus tumour and one with a mixed malignoma, both in the third stage of the disease. The remaining patients are alive and feel well. Except for three, all of them have been followed up for more than five years. PMID- 2557490 TI - Mineralocorticoid-induced increase in beta-adrenergic receptors of cultured rat arterial smooth muscle cells. AB - We have investigated the effect of mineralocorticoids on beta-adrenergic receptors in cultured arterial smooth muscle cells. Mineralocorticoid (aldosterone) treatment resulted in a significant increase in beta-adrenergic receptors measured by [3H]dihydroalprenolol (DHA) binding. This effect required at least 20 hours of incubation with aldosterone and was completely blocked by cycloheximide (10 micrograms/ml), indicating protein synthesis was required for this response. Aldosterone at the concentration range of 10(-8)-10(-6) M increased [3H]DHA binding, but was ineffective at 10(-9) M. Scatchard analysis of [3H]DHA binding revealed that the observed significant increase in binding was due to an increased number of binding sites (P less than 0.05), and that the affinity was unchanged. The aldosterone (1 x 10(-8) M) effect was completely blocked by the combination of RU 38486 (10(-6) M) and spironolactone (10(-7) M), but not by the glucocorticoid antagonist RU 38486 alone. While basal c-AMP levels were not changed by aldosterone (10(-6) M) treatment, the isoproterenol (10(-6) M) stimulated level of c-AMP was significantly higher in cells treated with aldosterone (P less than 0.05). We conclude that aldosterone, acting through the mineralocorticoid receptor, has a direct effect on arterial smooth muscle cells mediated through modulation of beta-adrenergic receptors of these cells. PMID- 2557492 TI - [Human papillomavirus types and cervical neoplasia]. AB - The pathological cells of the cervix uteri were examined for the presence of some types of human papilloma viruses (HPV) by DNA-hybridization and by the Southern blot method. Different degrees of neoplasia were found in the majority of these pathological changes of the uterus. The control group of patients included in the study had normal clinical, cytological and colposcopical results. Two parameters were identical both in the examined and the control patients. Types 6/11 were found in neither of the groups examined. The control patients had types 16/18 HPV in 8.3% of cases, while the frequency of these types was much higher in the examined group; it depended on the phase of the disease. These types were not detected in pathological lesions of CIN I; 40% of patients with CIN II had them, and 7.3% of patients with CIN III were positive to types 16/18 HPV. The majority (75%) of the examined samples of invasive neoplastic cervical lesions were positive to the presence of HPV 16/18. PMID- 2557493 TI - The Rochester Coronary Heart Disease Project: effect of cigarette smoking, hypertension, diabetes, and steroidal estrogen use on coronary heart disease among 40- to 59-year-old women, 1960 through 1982. AB - A population-based case-control study of coronary heart disease (CHD) risk in young women attributable to cigarette smoking, hypertension, diabetes, and steroidal estrogen use was conducted among residents of Rochester, Minnesota. All newly diagnosed cases of CHD (sudden unexpected death [SUD], N = 18; myocardial infarction [MI], N = 90; and angina, N = 133) among female Rochester residents 40 to 59 years of age during the years 1960 through 1982 were identified, and two community control subjects were matched for age and duration of community medical record. The overall adjusted odds ratio (OR) for the association between steroidal estrogen use and definite CHD (MI and SUD) was 0.6 (95% confidence interval [CI] = 0.2 to 1.3). Smoking (OR = 5.1; 95% CI = 2.3 to 11.6), hypertension (OR = 4.8; 95% CI = 2.3 to 10.2), and diabetes (OR = 8.4; 95% CI = 1.6 to 44.5) were strong risk factors for CHD events. If considered causal, cigarette smoking accounted for 64% of all MIs and SUDs in the community, hypertension accounted for 45%, and diabetes accounted for 13%. Although steroidal estrogen exposure reduced CHD among these women by 14%, giving steroidal estrogens to all women in this age group might reduce the population rates of MI by as much as 45%. PMID- 2557494 TI - Diagnostic interpretation of the intravenous tolbutamide test for insulinoma. AB - The plasma glucose and insulin responses to intravenous administration of tolbutamide in patients with insulinoma and healthy control subjects were compared to determine the sensitivity and specificity of the intravenous tolbutamide test for the diagnosis of insulinoma. The records of 406 healthy persons without concurrent disease known to affect glucose homeostasis or insulin concentrations and 41 patients with histologically confirmed insulinoma who underwent standard intravenous tolbutamide testing during the period from 1976 to 1986 were reviewed. The 5th percentile of the mean of plasma glucose levels at the 120-, 150-, and 180-minute points (G120-180) after injection of tolbutamide was 55 mg/dl for lean and 62 mg/dl for obese control subjects. With 95% specificity, the sensitivity of these criteria was 95% for lean and 100% for obese patients with insulinoma. Minimal differences were observed between men and women. At 95% specificity, the sensitivities were less for the ratio of the 180 minute plasma glucose to fasting plasma glucose level (64% in lean and 75% in obese patients), for mean of plasma insulin at the 120-, 150-, and 180-minute points (IRI120-180) (53% in lean and 36% in obese patients), for maximal insulin concentration (27% in lean and 31% in obese patients), for increase in insulin concentration above basal (18% in lean and 23% in obese patients), and for the combined criteria of the 5th percentile of G120-180 and the 95th percentile of IRI120-180 (47% in lean and 73% in obese patients).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557496 TI - Smoking, estrogen, and prevention of heart disease in women. PMID- 2557495 TI - Autoantibodies bind solubilized calcium channel-omega-conotoxin complexes from small cell lung carcinoma: a diagnostic aid for Lambert-Eaton myasthenic syndrome. AB - Serum autoantibodies found by radioimmunoassay in 27 of 52 patients with the Lambert-Eaton myasthenic syndrome (LES) bound specifically to a soluble omega conotoxin binding component of a voltage-gated Ca2+ channel (VGCC) complex extracted from small cell lung carcinoma (SCC). These antibodies were not found in 43 control patients with other neurologic diseases, including myasthenia gravis, peripheral neuropathies, and amyotrophic lateral sclerosis, or in 9 patients with endocrine autoimmunity, but they were found in 2 of 21 control patients with SCC without a history of LES, 1 of whom had severe autonomic neuropathy. Seropositivity was more frequent in patients with LES who had evidence of a primary lung cancer (76%) than in those with other neoplasms or without evidence of cancer (30%). Antigens extracted from SCC tumor lines derived from patients with and without LES and from a human neuroblastoma line yielded results that were highly correlated. A control extract of colonic carcinoma (derived from a patient with LES) yielded negative results. The data implicate a tumor-associated VGCC as the autoimmunizing stimulus in a subset of patients with LES and provide the first direct evidence that the VGCC complex in SCC is a target for some LES antibodies. The serologic test described should be a useful aid in diagnosing LES. PMID- 2557497 TI - Cellular signaling mechanisms common to the development and degeneration of neuroarchitecture. A review. AB - The present review examines the hypothesis that similar cellular signaling mechanisms are involved in neural development and in age- or disease-associated degeneration. It is hoped that approaching the problem of the regulation of brain structure from this perspective will spur future studies on the links between development, aging and disease. In order for functional neural circuitry to form, the component neurons must interact in highly specific ways. Growth factors and neurotransmitters constitute two major classes of intercellular signals that sculpt neuroarchitecture. These signals influence the neuronal growth cone behaviors which ultimately determine the details of neuritic form. In addition, growth factors and neurotransmitters can influence neuronal survival and synapse formation, and thereby determine both the presence of neurons within circuits and their specific connectivity patterns. Imbalances in growth factor and/or neurotransmitter systems may lead to neurodegeneration in aging and in specific neurodegenerative disorders such as Alzheimer's disease. Developmental, functional and pathological studies of excitatory amino acid neurotransmitters provide a compelling example of how a common intercellular signal can be involved in neuronal development, plasticity and degeneration. Intracellular signaling systems mediate neuroarchitectural responses to neurotransmitters and growth factors by altering the status of the cytoskeletal and vesicular substrates that are the basis of neuronal form. These signal transduction systems include ion channels and second messengers such as calcium, cyclic nucleotides and diacylglycerol. Cytoskeletal and vesicular substrates may be influenced directly by second messenger kinases, or indirectly via actions on the biosynthetic and degradative systems of the cell. Alterations in these various intracellular neuroarchitecture-regulating systems can lead to neurodegeneration. Taken together, the data presented here indicate that similar cellular and molecular mechanisms are involved in nervous system development, function, adaptive plasticity and degeneration. PMID- 2557498 TI - [Multifocal polyneuropathy with persistent conduction blockage. A new subset of chronic inflammatory polyneuropathies]. AB - Recently a subset of chronic demyelinating inflammatory polyneuropathies with asymmetrical involvement limited to upper limbs, at least at the onset, resembling a multifocal mononeuropathy has been described. Electrodiagnostic testing disclosed multifocal CB outside the common entrapment sites has been described. We report a 55 years old man with a 4 years history of paresis, numbness, fasciculations, myokymia, cramps and mild amyotrophy. Electrophysiological evaluation showed proximal multifocal conduction block and abundant spontaneous activity as fasciculations, myokymia and scarce denervation activity. The importance of taking into account this entity in the differential diagnosis of patients with suspected mononeuritis multiplex or motoneuron disease is emphasized. The nosologic place of this entity is also discussed. PMID- 2557499 TI - [Prognostically relevant factors in cancer of Vater's ampulla]. AB - In order to evaluate the relevant factors for prognosis for patients with cancer of Vater's papilla our own patient material between 1. 1. 1978 and 31. 12. 1987 was studied retrospectively. The influence of age, sex, duration of anamnesis, tumor size, tumor morphology and tumor stage on the prognosis were analyzed in 32 patients with cancer of Vater's papilla. The 5 year survival rate after Whipple procedure was 56.1%. Statistically significant factors for prognosis were tumor size, lymph node metastases and tumor stage. PMID- 2557500 TI - Management of the internal carotid artery in surgery of the skull base. AB - Management of the internal carotid artery in skull-base surgery can be a difficult problem when disease involves this vessel or resection is anatomically limited by it. The recent radiologic development of the detachable balloon catheter has permitted occlusion of the internal carotid artery in a controlled setting prior to any surgical procedure. An obvious prerequisite to using this technique is the demonstration of adequate collateral blood flow to the brain. Patients are evaluated with arteriography and temporary arterial balloon occlusion while monitoring physical signs and electroencephalography (EEG). Although usually performed preoperatively, internal carotid artery occlusion is needed intraoperatively on occasion. This essential adjuvant technique for the skull-base surgeon will be detailed along with its indications and limitations in 24 patients. PMID- 2557501 TI - Effects of preoperative embolization on glomus jugulare tumors. AB - The charts of 35 patients were reviewed retrospectively to determine the effects of embolization on glomus jugulare tumors. Eighteen patients underwent embolization; 17 did not. All tumors were removed with a type A infratemporal fossa approach. The charts were evaluated for operative blood loss, operative time, length of hospitalization, new postoperative cranial nerve deficits and recurrence of tumor. Embolized patients had significantly less operative blood loss and operative time. Embolization did not affect length of hospitalization or reduce the incidence of new postoperative cranial nerve deficits. The surgeon's experience also plays a central role in reducing operative blood loss and operative time. The potential risks of embolization must be considered in the treatment regimen of glomus jugulare tumors. PMID- 2557502 TI - Comparative study of suture and laser-assisted anastomoses in rat sciatic nerves. AB - Conventional suture repair of peripheral nerves results in a fibrotic reaction that is detrimental to nerve regeneration. As an alternative procedure known as "laser-assisted" repair, a laser can be used, along with a reduced number of sutures, to reanastomose served peripheral nerves. To explore the long-term implications of this technique, the right sciatic nerves of Sprague-Dawley rats were surgically cut and reanastomosed either by means of four epineurial sutures or two epineurial sutures and CO2 laser welds. Tensile strength, electrophysiology, histology, and functional studies were performed up to 11 months postoperatively. Tensile strength measurements indicate no long-term disadvantage with the laser-assisted technique, although the short-term tensile strength is lower than with conventional suture repair. The conduction velocities of the repaired nerves were similar for both techniques; however, laser-assisted repaired nerves were found to have lower stimulation thresholds and reduced branching compared to the suture repaired nerves. The measured functional recovery was similar for both repair techniques. PMID- 2557503 TI - Electron spin resonance investigation of laser and heated metal-tip-induced free radical formation in various tissues. AB - The short-lived free radical formation accompanying laser irradiation and laser heated metal tip contact was examined using electron spin resonance (ESR) spin trap methodology. Various tissues (canine myocardium, human aorta, liver, and spleen) were irradiated by argon-ion (continuous wave [CW] and pulse) and YAG (CW) lasers employing both naked fiber and a laser-heated metal tip. All showed the formation of primarily carbon-centered, not oxygenated, free radicals, together with some minor unidentified species. This implies that the backbones of amino acids, lipids, or other biological building blocks are cleaved during the irradiation or thermal treatment. Different tissues produce similar radicals but with different amounts when irradiated by argon-ion, YAG, and laser-heated metal tip sources. The amount of the free radicals formed depends on the laser power (within 5-15 W). Compared to the naked fiber, the laser-heated metal tip shows the generation of at least twice the amount of free radicals. The possible relationship of the free radical formation to tissue injury is briefly discussed. PMID- 2557504 TI - The envelope glycoprotein of HIV-1 may have incorporated the CD4 binding site from HLA-DQ beta 1. AB - An hypothesis is presented which states that the increased binding for CD4 by the envelope glycoprotein (gp120) from HIV-1 compared with that from HIV-2 is due to the env gene from HIV-1 having at some stage incorporated exon 2 of the gene coding for the beta subunit of a class II MHC protein, possibly HLA-DQ, which contains part of the CD4 binding site. Evidence is presented from amino acid sequence analysis and consideration of putative binding residues from gp120 and HLA-DQ. PMID- 2557505 TI - Modification of the positive inotropic effects of catecholamines, cardiac glycosides and Ca2+ by the orally active male contraceptive, gossypol, in isolated guinea-pig heart. AB - Gossypol is an orally active male contraceptive with cardio-depressant side effects. To understand the mechanism of its cardiac actions, the interaction of gossypol with positive inotropic drugs was examined in isolated atrial muscle preparations obtained from guinea-pig heart. Gossypol delayed the onset of arrhythmias caused by digoxin. In the presence of gossypol, the positive inotropic effect of isoproterenol declined rapidly, and the effect of isoproterenol to increase tissue cyclic AMP concentrations was smaller. Pretreatment of atrial muscle with the combination of gossypol and isoproterenol markedly reduced effects of isoproterenol on developed tension and cyclic AMP concentrations when these effects were tested after the washout of the first dose of isoproterenol. These effects, however, were not specific to isoproterenol. The gossypol-isoproterenol pretreatment reduced the positive inotropic effect of ouabain or extracellular Ca2+. These results indicate that gossypol has pharmacodynamic interactions with several positive inotropic agents that are known to enhance developed tension by increasing intracellular Ca2+ transients. PMID- 2557506 TI - Subcellular compartmentalization of cAMP-dependent protein kinase regulatory subunits during palate ontogeny. AB - Mammalian palatal ontogeny involves epithelial-mesenchymal interactions, cell differentiation, and cell movements. These events occur on days 12, 13, and 14 of gestation in the C57BL/6J mouse embryo. During this period intracellular cAMP levels and cAMP-dependent protein kinase (cAMP-dPK) levels in the palate transiently elevate. Cyclic AMP activates cAMP-dPK by binding primarily to two types of regulatory subunits of this enzyme, designated as RI and RII. To assess whether differential compartmentalization of the regulatory subunits occurs during palatal ontogeny, cytosolic, nuclear, and particulate fractions were prepared from day 12, 13, and 14 embryonic maxillary and palatal tissue. After photo-affinity labeling of each fraction with 8-azido [32P] cAMP, SDS-PAGE, and autoradiography, autoradiograms were analyzed densitometrically. The RI isoform predominated in the nuclear and particulate fractions on all three developmental days; whereas RII predominated in the cytosolic fractions. Thus, differential compartmentalization of cAMP-dPK may be a means by which cAMP dependent responses are regulated during palatogenesis. PMID- 2557507 TI - Amoxapine inhibition of GABA-stimulated chloride conductance: investigations of potential sites of activity. AB - Amoxapine inhibits GABA-stimulated chloride conductance by acting on the GABAA receptor chloride-ionophore complex which can be studied using membrane vesicles prepared from rat cerebral cortex. Amoxapine produces a right shift in the GABA concentration-response curve for the stimulation of 36Cl- uptake into these vesicles with no apparent change in the maximum response. Schild analysis of these data gave a pA2 value of 5.52 with a slope of 0.79. Amoxapine inhibits the binding of the GABAA receptor selective antagonist [3H]SR 95531 with an IC50 value of 3.45 microM and a pseudo Hill coefficient of 0.83. In contrast, 10 microM amoxapine inhibits [3H]flunitrazepam binding by less than 25% while the benzodiazepine antagonist Ro 15-1788 reduces the amoxapine inhibition of GABA stimulated chloride conductance only at high concentrations. These data suggest that amoxapine does not inhibit chloride conductance by acting as a benzodiazepine inverse agonist and either acts directly on the GABAA receptor as an antagonist or blocks GABA activity at a site closely coupled to it. The ability of amoxapine to inhibit GABA-stimulated chloride conductance is a likely explanation for its proconvulsant activity observed at high doses. PMID- 2557508 TI - Morphine-induced desensitization and down-regulation at mu-receptors in 7315C pituitary tumor cells. AB - Pituitary 7315c tumor cells maintained in culture were treated with varying concentrations of morphine from 10 nM to 300 microM, for periods of five or forty eight hours. The ability of the mu-opioid receptor agonist, DAMGO, to inhibit forskolin-stimulated adenylyl cyclase in washed membrane preparations from the treated cells was compared with its activity in membranes from cells incubated in the absence of added morphine. In the same membrane preparations, the number and affinity of mu-opioid receptors was estimated by measurements of [3H]diprenorphine binding. After 5 hr of treatment with morphine concentrations of 100 nM or higher, a significant reduction in inhibition of adenylyl cyclase by DAMGO was observed. Little further loss of agonist activity was observed when the incubations were extended to 48 hr. After 5 hr of morphine treatment, there was no change in either the number of receptors, or their affinity for [3H]diprenorphine. However, after 48 hr of morphine treatment, greater than 25% reductions in receptor number were apparent with morphine pretreatment concentrations of 10 microM or higher. These results suggest that opioid tolerance in this system is primarily associated with a reduced ability of agonist-occupied receptor to activate the effector system. Receptor down regulation was not necessary for loss of agonist response, although a reduction in receptor number occurred after exposure to high concentrations of morphine for periods longer than 5 hr. PMID- 2557509 TI - Manganese attenuates secretagogue-mediated phospholipid hydrolysis in AR42J cells. AB - The effects of cholecystokinin octapeptide (CCK8), bombesin and manganese (Mn2+) on phosphatidylinositol-4,5-bisphosphate (PIP2) hydrolysis were studied in AR42J cells. One-half maximal stimulation of inositol monophosphate (InsP1) accumulation occurred at either 5 nM CCK8 or 5 nM bombesin, and maximal stimulation occurred at 30 nM for each agonist. Mn2+ did not alter basal PIP2 hydrolysis. However, addition of Mn2+ 5 min prior to stimulation with either CCK8 or bombesin for 60 min significantly attenuated [3H]InsP1 accumulation. Following brief periods of incubation with CCK8 (15 sec) Mn2+ significantly reduced inositol tris- and tetrakisphosphate accumulation. These data suggest that Mn2+ may participate in the regulation of CCK8- and bombesin-mediated generation of phosphoinositides. PMID- 2557511 TI - Angiotensin II increases diacylglycerol in calf adrenal glomerulosa cells by activating de novo phospholipid synthesis. AB - Effects of angiotensin II (AII) on diacylglycerol (DAG) synthesis were examined in calf adrenal glomerulosa cells. AII provoked rapid increases in [3H]glycerol labeling and content of DAG. Effects on [3H]glycerol-labeling of DAG were observed both in cells prelabeled with [3H]glycerol for 60 minutes, and when AII and [3H]glycerol were added simultaneously. Increases in [3H] DAG labeling were associated with increases in total glycerolipid labeling, and in simultaneous addition experiments, were preceded by increased [3H] phosphatidic acid (PA) labeling. Labeling of glycerol-3-PO4, on the other hand, was not increased by AII, suggesting that increases in lipid labeling were not due to prior increases in precursor specific activity. ACTH, which does not increase the hydrolysis of inositol-phospholipids appreciably in this tissue, provoked increases in content and [3H]glycerol-labeling of DAG, which were only slightly less than those provoked by AII. Thus, part of the AII-induced increase in DAG may also be derived from sources other than inositol-phospholipids. Moreover, AII-induced increases in DAG appear to be at least partly derived from increased de novo synthesis of PA. PMID- 2557510 TI - Calcitonin inhibits accumulation of cyclic AMP in stimulated peritoneal macrophages from normal rats but not from osteopetrotic (incisors-absent) littermates. AB - Macrophages and osteoclasts derive from related cell lines. In osteopetrotic mutants the function of osteoclasts is greatly reduced compared to that in normal animals or children and macrophage function is variably affected depending upon the mutation. To further explore macrophage function in osteopetrosis we examined the regulation of cyclic AMP production in macrophages from mutants and normal littermates of the osteopetrotic stock incisors-absent (ia) in the rat. Surface stimulation by latex particles of elicited peritoneal macrophages from normal or osteopetrotic (ia) mutant rats caused an identical increase in the accumulation of cyclic AMP. This effect was inhibited in normal animals by coincubation of macrophages with calcitonin (CT) but this inhibition was either absent or less marked in macrophages from mutant littermates. In contrast to human monocytes preincubation of rat macrophages with pertussis toxin did not relieve this inhibition. This implies that rat peritoneal macrophages respond to CT by a different mechanism. These results demonstrate altered macrophage function in osteopetrotic animals and may be functionally related to the reduced CT binding previously described in ia osteoclasts. Furthermore, the coexistence of reduced function of macrophages and osteoclasts in the ia mutation suggests that macrophages and osteoclasts share a common progenitor. PMID- 2557512 TI - Corticotropin-releasing factor and adrenocorticotropin stimulate ciliary motility in rabbit tracheal epithelium. AB - To assess the effects of corticotropin-releasing factor (CRF) and adrenocorticotropin (ACTH) on airway ciliary activity, we measured ciliary beat frequency (CBF) by a photoelectric method in response to these peptides in cultured rabbit tracheal explants. When cumulatively added, both CRF and ACTH increased CBF in a dose-dependent fashion. Treatment of tissues with Ca2+-free medium or nifedipine abolished the effect of CRF but not of ACTH. The CRF- and ACTH-induced ciliostimulations were not affected by indomethacin or autonomic antagonists, but were attenuated by nordihydroguaiaretic acid and by their receptor antagonists, alpha-helical CRF (9-41) and ACTH (7-38). Intracellular cyclic AMP levels were significantly increased by CRF and ACTH. These results suggest that CRF and ACTH stimulate airway ciliary motility through the activation of adenylate cyclase and lipoxygenase by binding to their specific receptors, where the effect of CRF may be triggered by Ca2+ influx. PMID- 2557513 TI - A randomized, controlled dose response study of intravenous sodium diethyldithiocarbamate in patients with advanced human immunodeficiency virus infection. AB - Sodium diethyldithiocarbamate (Imuthiol, DTC) has previously been observed to promote T-cell maturation in animal models and to reduce lymphadenopathy and improve survival in a murine AIDS model. In addition, several clinical studies have suggested that one dosage regimen may be active in patients with HIV infection. We conducted a randomized, controlled dose response study of intravenous DTC in patients with the acquired immunodeficiency syndrome (AIDS) and AIDS-related complex (ARC). Drug associated toxicities included gastrointestinal upset, burning at the infusion site, metallic taste, sneezing, confusional states, hyperactivity, delusional thinking, and myoclonus. Toxicity was ameliorated by dose reduction. The maximally tolerated dose varied for individual patients from 200 mg/m2 weekly to 800 mg/m2 twice weekly. No myelosuppression was observed. In patients with greater than 200 CD4+ cells/uL, a statistically significant reduction of lymphadenopathy occurred; whereas no beneficial effects were observed in patients with less than 200 CD4+ cells/uL. Improvement in symptom score and stabilization of CD4+ count also occurred in the treated group, although these trends did not reach statistical significance. Further controlled clinical trials of DTC in earlier HIV infection are warranted. PMID- 2557514 TI - Inverse regulation of alpha-2 and beta-2 adrenoceptors in salt-sensitive hypertension: an hypothesis. AB - A high salt diet leads to up-regulation of alpha-2 adrenoceptors and down regulation of beta-2 adrenoceptors in normotensive subjects. Although the increase in blood pressure with a high salt diet is not related to the magnitude of the alpha-2 or beta-2 adrenoceptor changes alone, it is related to the increase in the ratio of the receptor changes (operative alpha/beta adrenoceptor ratio). An increase in the operative alpha/beta adrenoceptor ratio with a high salt intake results in vasoconstriction and reduced vasodilatation at resistance vessels, as well as increased renal proximal tubular sodium reabsorption. An influence of heredity on this relationship is supported by four lines of evidence: 1) salt-sensitivity of blood pressure occurs predominantly in subjects with a family history of hypertension; 2) studies in twin children document the influence of genetic variance on salt-sensitivity of blood pressure; 3) subjects with a family history of hypertension have a significantly lower salivary sodium concentration and an altered urinary sodium excretion after salt loading compared to subjects with no such history; 4) salt-sensitivity of blood pressure may be associated with specific genetic markers. On the basis of these observations, we propose the hypothesis that enhanced inverse alpha-beta-adrenoceptor regulation in response to a high salt intake may be responsible for salt sensitivity in the normal population, and may contribute to the development of essential hypertension in susceptible individuals. This alteration is likely to be genetically mediated. PMID- 2557515 TI - Distribution of kappa opioid receptors in the brain of young and old male rats. AB - The experiments to be described have been designed in order to: (a) provide new information on the concentrations of opioid kappa receptors in different regions of the brain of the male rats; and (b) to analyze whether the density of brain kappa receptors might be modified by the process of aging. The concentration of kappa receptors was investigated in the hypothalamus, amygdala, mesencephalon, corpus striatum, hippocampus, thalamus, frontal poles, anterior and posterior cortex collected from male rats of 2 and 19 months of age. 3H-bremazocine (BRZ) was used as the ligand of kappa receptors, after protection of mu and delta receptors respectively with dihydromorphine and d-ala-d-leu-enkephalin. The results obtained show that: (1) in young male rats, the number of kappa opioid receptors is different in the various brain areas examined: the hypothalamus and the striatum have a concentration of kappa binding sites which is significantly higher than that found in the mesencephalon and in the amygdala; much lower concentrations of kappa binding sites have been found in the thalamus, the frontal poles, the hippocampus, the anterior and posterior cerebral cortex. (2) Aging exerts little influence on the number of kappa receptors in the majority of the brain structures considered. However in the amygdala and in the thalamus the number of kappa receptors was increased in old animals. To the authors' knowledge, the data here presented are the first ones which suggest that age may increase rather than decrease the number of neurotransmitter receptors in the brain. PMID- 2557516 TI - Failure to find Ca2(+)-dependent proteinase (calpain) activity in a plant species, Elodea densa. AB - Five and nine-tenth kg of Elodea densa (Anacharis), a common aquarium plant, was extracted, and the extract was subjected to column chromatographic procedures that successfully purify the two Ca2(+)-dependent proteinases (calpains) and their protein inhibitor (calpastatin) from a variety of animal tissues. Although these procedures purified a protein having 55- and 16-kDa polypeptides, neither this protein nor any of the other chromatographic fractions contained detectable proteinase or calpastatin activity. Moreover, the purified 55- and 16-kDa polypeptides did not react on immunoblots with polyclonal antibodies that were monospecific for the calpains or calpastatin. We conclude that Elodea densa contains no calpain nor calpastatin at the level of 4 micrograms per g plant protein (1 part per 250,000), which was the sensitivity of our assay. PMID- 2557518 TI - Platelet prostaglandin E1 hyposensitivity in schizophrenia: reduction of prostaglandin E1- or forskolin-stimulated cyclic AMP response in platelets. AB - Cyclic 3',5'-adenosine monophosphate (cAMP) formation via prostaglandin E1 (PGE1) or forskolin-stimulation were determined in washed intact platelets from 32 schizophrenic patients and 30 normal controls. Regarding basal cAMP levels in the platelets, there were no differences between schizophrenic patients and normal controls. Both PGE1-and forskolin-stimulated cAMP response reduced in platelets from schizophrenics compared with normal controls. These results suggested that platelets in schizophrenics were impaired not only in the adenylate cyclase unit per se but also extensively in the cAMP generating system coupled to a PGE1 receptor. PMID- 2557517 TI - Supersensitivity of beta-adrenoceptor coupled adenylate cyclase in pulmonary tissue of the spontaneously hypertensive rat. AB - Basal adenylate cyclase activity was similar in plasma membranes prepared from the lungs of 12 week old spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY). However, sensitivity to Gpp[NH]p, isoproterenol plus GTP or Gpp[NH]p was significantly greater in the SHR. Beta-receptor density measured by [3H]DHA binding was unaltered. The dissociation constant, Kd, revealed a significantly greater binding affinity of the radioligand in the SHR (6.23 +/- 0.45 nM) compared with the WKY (8.53 +/- 0.82 nM). Activity of Gs was assessed by complementing S49 cyc- acceptor membranes with lung cholate extract. Basal activity of the reconstituted system was decreased 43% in the SHR. However, sensitivity to NaF, Gpp[NH]p, and isoproterenol plus Gpp[NH]p was significantly elevated. These data suggest that desensitization of the adenylate cyclase complex is not a generalized response to chronic hypertension. A tissue specific increase in sympathetic drive appears to be responsible for the lowered concentration of cardiac beta-adrenoceptors in the SHR. In contrast, both indirect and direct evidence indicate an enhanced functional sensitivity of pulmonary Gs in the hypertensive rats. PMID- 2557519 TI - Insulin control of cyclic AMP phosphodiesterase. AB - Cyclic AMP phosphodiesterase (PDE) is an enzyme involved in cellular homeostasis of cyclic AMP. It exists as multiple isozymes in cells, but only the high affinity, membrane-bound isozyme is sensitive to hormonal modulation. Several isozymes or isoforms of the low Km PDE have been detected. Data suggest that several mechanisms exist for hormonal modulation of PDE. Activity of the low Km PDE species may be modulated by phosphorylation/dephosphorylation, phospholipid substrate concentration, insulin second messenger, cyclic GMP, guanine nucleotide binding proteins, calmodulin, or aggregation/disaggregation of monomeric forms. Modulation of PDE isoforms by different hormones may be through different regulatory components or mechanisms. PMID- 2557520 TI - Ligand binding and internalization by the rat hepatic asialoglycoprotein receptor does not generate polyphosphoinositide derived second messengers. AB - Recent studies suggest that protein kinase C and, thus, possibly the rate of inositol phospholipid hydrolysis may regulate the function and distribution of the asialoglycoprotein (or galactosyl) receptor on isolated rat hepatocytes (Takahashi et al., Biochem. Biophys. Res. Commun., 1985, 126, 1054; Fallon and Schwartz, J. Biol. Chem., 1986, 261, 15081). We have studied the effects of asialoorosomucoid (ASOR) on the hydrolysis of [32P]-inositol phospholipids in isolated rat hepatocytes. When internalization of ASOR is maximal at 310 molecules/cell/sec, there is neither a decrease in the amount of [32P] phosphatidylinositol-4,5-bisphosphate (PIP2) nor an increase in [32P] phosphatidic acid (PA) up to 30 min after stimulation. On the other hand, 10(-6)M vasopressin, which was used as a positive control, caused a 35-40% decrease in the level of [32P]-PIP2 and a 70-80% increase in [32P]-PA within 30 sec. Addition of orosomucoid or ASOR, even at concentrations 1000-times the Kd, did not change the levels of any of the six phospholipids tested. Similarly, addition of ASOR did not increase the levels of soluble [3H]-inositol phosphates, whereas vasopressin caused a 6-fold increase in [3H]-inositol-1,4-diphosphate (IP2) and a 4-fold increase in [3H]-inositol-1,4,5-triphosphate (IP3) in isolated rat hepatocytes prelabeled with [3H]-inositol. We conclude that the receptor mediated endocytosis of asialoglycoproteins by rat hepatocytes does not stimulate hydrolysis of the inositol phospholipids. PMID- 2557521 TI - [Salivary gland function after radioiodine therapy of thyroid cancer]. PMID- 2557522 TI - [Scintigraphy in detecting suppurative-septic foci in the abdominal cavity and retroperitoneal space]. AB - Positive results of the use of 99mTc-pyrophosphate in 91.9% of patients with pyoseptic foci of various sites prove a regular character of increased accumulation of this agent in abscesses and permit recommending this method as a screening test in their indefinite localization. If the foci are localized in the liver, positive scintigraphy should be supplemented by negative scintigraphy of the liver, increasing the diagnostic value of radionuclide methods. PMID- 2557523 TI - Interaction of anti-idiotypic antibodies with membrane receptors: practical considerations. PMID- 2557524 TI - Immunocytochemistry of receptors using anti-idiotypic antibodies. PMID- 2557525 TI - Production and properties of antimorphine anti-idiotypic antibodies and their antiopiate receptor activity. PMID- 2557526 TI - Production and characterization of anti-idiotypic antiopioid receptor antibodies. PMID- 2557528 TI - Production and properties of site-specific antibodies to synthetic peptide antigens related to potential cell surface receptor sites for rhinovirus. PMID- 2557527 TI - Development and use of antireceptor antibodies to study interaction of mammalian reovirus type 3 with its cell surface receptor. PMID- 2557529 TI - Production and use of synthetic peptide antibodies to map region associated with sodium channel inactivation. PMID- 2557530 TI - Auer rods and myeloperoxidase: an approach for investigating changes at the molecular level in acute myeloid leukaemia (AML). AB - Auer rods are cytoplasmic inclusions found only in the leukaemic cells of some cases of acute myeloid leukaemia (AML). The ultrastructure of Auer rods is different in different sub-types of the disease and it is suggested here that this may be a consequence of the Auer rods consisting of different crystalline forms of the enzyme myeloperoxidase. There is statistical evidence to indicate that patients with Auer rods have a more favourable prognosis than those without. It is proposed that an investigation of the factors that determine the form, if any, of the crystallisation of myeloperoxidase will lead to a better understanding of the molecular events underlying the different types of AML. In the long term this may in turn lead to a therapeutic advance. PMID- 2557532 TI - [Intestinal polyposis in children. Description and differential diagnosis of 2 cases]. AB - The case of a 21-month-old girl with lymphoid nodular hyperplasia and the case of a 9-year-old girl with familial adenomatous polyposis are described. Both patients presented rectal bleeding. A defect of secretory IgA was found in the first patient. In both cases diagnosis was based on medical history, barium enema with aircontrast technique, colonscopy, and endoscopic biopsy. Differential diagnosis and diagnostic procedures in these two diseases are discussed. PMID- 2557531 TI - Evidence for the presence of a benzodiazepine receptor binding substance in cerebrospinal fluid of a rabbit model of hepatic encephalopathy. AB - Based on the reversal of hepatic encephalopathy in animal models with administration of specific benzodiazepine receptor antagonists, it has been postulated that this syndrome may be mediated by an endogenous benzodiazepine like compound. In this study using a radio-receptor assay, evidence for the existence of this substance has been demonstrated in cerebrospinal fluid but not sera of rabbits with hepatic encephalopathy due to galactosamine-induced hepature failure. Cerebrospinal fluid from rabbits with hepatic encephalopathy caused 36.1 +/- 5.03% displacement of 3H-Ro 15-1788 specific binding to cortical benzodiazepine receptors, compared to 11.7 +/- 0.76% in control animals (P less than 0.01). The benzodiazepine receptor binding activity has been shown to behave as a competitive inhibitor of radiolabeled benzodiazepine receptor binding. The finding of endogenous benzodiazepine binding activity affords a potential explanation for the amelioration of hepatic encephalopathy in this model with the administration of benzodiazepine receptor antagonists. PMID- 2557533 TI - Spermine enhances binding to the glycine site associated with the N-methyl-D aspartate receptor complex. AB - Spermine enhanced strychnine-insensitive [3H]glycine binding 3-fold with an EC50 of 27 +/- 3.1 microM. Spermidine and putrescine were without effect, whereas the ethylenediamine analog of spermine had an intermediate effect. Eadie-Hofstee analysis revealed that spermine increased the affinity of glycine for its receptor without a significant change in receptor density. This effect persisted in the presence of glycine or N-methyl-D-aspartate receptor antagonists. Furthermore, spermine produced a leftward shift in the IC50 of glycine agonists in displacing [3H]glycine binding, without altering the IC50 for glycine antagonists. These data indicate that spermine interacts with the glycine receptor through a novel binding site and, further, that spermine can be used to discriminate glycine agonist and antagonist binding. PMID- 2557534 TI - Site-directed mutagenesis of m1 muscarinic acetylcholine receptors: conserved aspartic acids play important roles in receptor function. AB - Muscarinic acetylcholine receptors contain a region encompassing the second and third transmembrane domains that is rich in conserved aspartic acid residues. To investigate the role of four conserved aspartic acids at positions 71, 99, 105, and 122 in muscarinic receptor function, point mutations in the rat m1 muscarinic receptor gene were made that converted each Asp to Asn, and wild type or mutant genes were stably expressed in Chinese hamster ovary cells that normally lack muscarinic receptors. Substitution of Asp71 or Asp122 with Asn produced mutant receptors that displayed high affinity for carbachol but decreased efficacy and potency, respectively, in agonist-induced activation of phosphoinositide hydrolysis, suggesting that these residues may mediate receptor-GTP binding protein interactions. Substitution of Asp99 or Asp105 with Asn produced marked decreases in ligand binding affinities and/or covalent incorporation of [3H] propylbenzilylcholine mustard, suggesting that these residues may be involved in receptor-ligand interactions. PMID- 2557535 TI - Inhibition of epidermal growth factor receptor tyrosine kinase activity in A431 human epidermoid cells following psoralen/ultraviolet light treatment. AB - One of the more prominent clinical treatments for skin diseases such as psoriasis and vitiligo involves the use of a combination of psoralens and UV light, a procedure referred to as PUVA chemotherapy. This drug regimen markedly alters epidermal cell growth and differentiation. In many cell types, an early cellular event following treatment of cells with PUVA is inhibition of binding of epidermal growth factor (EGF) to its receptor. To examine the mechanism underlying this effect, we used A431 cells, a human epidermal cell line known to express large numbers of EGF receptors. We found that exposure of A431 cells to PUVA caused a dramatic inhibition of EGF-stimulated EGF receptor tyrosine kinase activity. Inhibition required intact cells and did not appear to be mediated by protein kinase C, because this inhibition was apparent in cells in which the enzyme was down-regulated by phorbol ester pretreatment and in cells treated with inhibitors of protein kinase C. Inhibition of tyrosine kinase activity by PUVA was distinct from other inhibitors of EGF receptor function in that it was associated with a rapid increase in the amount of phosphate incorporated into serine residues of the EGF receptor. This suggested that PUVA-induced serine phosphorylation may mediate EGF receptor kinase activity. These results demonstrate that alterations in EGF receptor function may contribute to the therapeutic efficacy of PUVA in photo-chemotherapy. PMID- 2557536 TI - The psychotomimetic drug phencyclidine labels two high affinity binding sites in guinea pig brain: evidence for N-methyl-D-aspartate-coupled and dopamine reuptake carrier-associated phencyclidine binding sites. AB - Numerous studies have now demonstrated that a binding site for the psychotomimetic drug phencyclidine (PCP) exists within the receptor channel complex for the excitatory amino acid neurotransmitter glutamate, specifically the glutamate receptor selectively activated by N-methyl-D-aspartate (NMDA). Several lines of evidence support the hypothesis that all PCP receptors in rat brain are associated with the NMDA receptor complex. In the present study, we reexamine this hypothesis. We report that the PCP analog [3H]1-[1-(2 thienyl)cyclohexyl]piperidine [( 3H]TCP) labels two high affinity binding sites in membranes prepared from guinea pig brain site 1 (Kd = 14.1 nM, Bmax = 631 fmol/mg of protein) and site 2 (Kd = 46.5 nM, Bmax = 829 fmol/mg of protein). (+) 5-Methyl-10 11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate bound to site 1 with high affinity (Kl = 3.2 nM) and to site 2 with low affinity (Kl = 5208 nM). The order of potency of drugs for inhibiting [3H]TCP binding to site 1 correlated with their ED50 values for inhibition of NMDA-mediated responses reported in the literature, whereas the order of potency of drugs for inhibiting [3H]TCP binding to site 2 correlated with their ED50 values for inhibition of [3H]dopamine reuptake reported in the literature. Kinetic experiments demonstrated that glutamate, 2-amino-7-phosphonoheptanoic acid, and Mg2+ modulated [3H]TCP binding to site 1 but not site 2. Preincubation of guinea pig striatal membranes with varying concentrations of the high affinity dopamine reuptake inhibitors N-[1-(2-benzo(b)thiophenyl)cyclohexyl]piperidine and 1-[2 [bis(4-fluorophenyl)methoxy]ethyl]-4-[3- phenylpropyl]piperazine caused a wash resistant inhibition of [3H]TCP binding to site 2 but not site 1. Taken collectively, these data demonstrate the existence of a high affinity PCP binding site associated with the dopamine reuptake carrier and raise the possibility that the therapeutic and psychotomimetic effects of PCP in humans are separable and mediated via different binding sites. PMID- 2557537 TI - Serotonin depletion unmasks serotonergic component of [3H]dihydroalprenolol binding in rat brain. AB - Selective lesions of serotonin neurons or inhibition of serotonin synthesis results in an increase in the number of [3H]dihydroalprenolol binding sites in several areas of rat brain. Previously, this increase in binding sites was interpreted as an increase in beta-adrenergic receptors. However, the lesion induced increase in [3H]dihydroalprenolol is not accompanied by an increase in isoproterenol- or norepinephrine-stimulated cyclic AMP production. The increased binding of [3H]dihydroalprenolol is blocked by the addition of serotonin but not by the addition of norepinephrine or dopamine to the assays. Furthermore, the addition of metergoline, a serotonin antagonist, also blocked the increase in lesioned tissues. Thus, the lesion-induced increase in [3H]dihydroalprenolol binding appears to represent an increase in serotonergic binding sites. Among drugs with some selectivity for serotonin-1 receptor subtypes, trifluoromethylphenylpiperazine and RU-24929 were as effective as serotonin in blocking lesion-induced increases in [3H]dihydroalprenolol binding. However, 8 hydroxy-N,N-dipropyl-2-aminotetralin (8-OH-DPAT), spiperone, and mesulergine were either much less effective than serotonin or completely ineffective. Radioligand binding to serotonin-1A and serotonin-1B sites with [3H]8-OH-DPAT and 125I cyanopindolol, respectively, after lesions of serotonin axons or depletion of serotonin was not increased, despite a marked increase in [3H]dihydroalprenolol binding in the same tissues. When tissues from control rats or rats with serotonin lesions were preincubated at 37 degrees for 10 min to remove endogenous serotonin bound to receptors, the binding of [3H]dihydroalprenolol in controls was increased to the level seen in lesioned tissues. Thus, [3H]dihydroalprenolol binds primarily to beta-adrenergic receptors in control membranes that are not preincubated; however, either preincubation of control tissues or serotonin depletion unmasks a serotonin-1 receptor subtype to which [3H]dihydroalprenolol binds in addition to the beta-adrenergic receptor. PMID- 2557538 TI - Identification of a signal transduction response sequence element necessary for induction of a Dictyostelium discoideum gene by extracellular cyclic AMP. AB - The signal transduction pathways that lead to gene induction are being intensively investigated in Dictyostelium discoideum. We have identified by deletion and transformation analysis a sequence element necessary for induction of a gene coding for uridine diphosphoglucose pyrophosphorylase (UDPGP1) of D. discoideum in response to extracellular cyclic AMP (cAMP). This regulatory element is located 380 base pairs upstream of the transcription start site and contains a G+C-rich partially palindromic sequence. It is not required for transcription per se but is required for induction of the gene in response to the stimulus of extracellular cAMP. The cAMP response sequence is also required for induction of the gene during normal development. A second A+T-rich cis-acting region located immediately downstream of the cAMP response sequence appears to be essential for the basal level of expression of the UDPGP1 gene. The position of the cAMP response element coincides with a DNase I-hypersensitive site that is observed when the UDPGP1 gene is actively transcribed. PMID- 2557539 TI - Processing of 9E3 mRNA and regulation of its stability in normal and Rous sarcoma virus-transformed cells. AB - We studied the expression of 9E3 mRNA, which is known to be induced in chicken embryo fibroblasts by p60v-src activity and by serum. In addition to full-length 9E3 mRNA, we identified several smaller RNAs that hybridized with 9E3 cDNA. One of these RNAs hybridized with a 5' 9E3 cDNA probe but not with a 3' cDNA probe. The other hybridized with a 3' cDNA probe but lacked 5' sequences, including the entire 9E3 coding region. Only the latter RNA was polyadenylylated, as determined by RNase H digestion in the presence of oligo(dT). The level of the small RNAs increased after treatment with cycloheximide and actinomycin D, indicating that the small RNAs were produced by processing of preexisting transcripts. The derivation of the small RNAs from 9E3 mRNA rather than from a related gene was confirmed by S1 nuclease analysis. The 3' terminus of the 5' RNA and the 5' terminus of the 3' RNA mapped to the same position, which suggested that the small RNAs were formed by endonucleolytic cleavage of 9E3 mRNA at a specific site in the 3' noncoding region. We also found that the stability of 9E3 mRNA was increased after serum stimulation and was greater in Rous sarcoma virus transformed than in uninfected cells. The relative amount of the small RNAs as compared with the full-length transcript was greatest under conditions in which the full-length transcript was least stable. These data suggest that site specific endonucleolytic cleavage regulates the stability of 9E3 mRNA. PMID- 2557540 TI - Activation of yeast polymerase II transcription by herpesvirus VP16 and GAL4 derivatives in vitro. AB - Fusion proteins known to activate transcription in vivo were tested for the ability to stimulate transcription in vitro in a recently developed Saccharomyces cerevisiae RNA polymerase II transcription system. One fusion protein, whose activation domain was derived from the herpesvirus transcriptional activator VP16, gave more than 100-fold stimulation in the in vitro system. The order of effects of the various proteins was the same for transcription in vitro and in vivo, suggesting that the natural mechanism of activation is preserved in vitro. PMID- 2557541 TI - UV-induced early-domain binding factor as the limiting component of simian virus 40 DNA amplification in rodent cells. AB - UV radiation and other carcinogenic agents induce an increase in DNA-binding activity to the early domain of the simian virus 40 (SV40) minimal origin in both SV40-permissive and SV40-nonpermissive cells. The increase is due to posttranslational modification of a preexisting protein, since it occurs in the presence of cycloheximide or anisomycin. Binding of this factor is an absolute requirement for the UV-induced SV40 DNA amplification in Co631 cells in vivo. A synthetic double-stranded oligonucleotide covering the early domain sequence totally blocked the UV-induced amplification in competition experiments. Point mutants of the sequence and unrelated oligonucleotides which could not bind the factor also did not block SV40 amplification. Inhibitors of protein synthesis caused an immediate increase of both early-domain factor activity (perhaps by prolonging mRNA half-life for the factor or for a modifying enzyme) and DNA amplification. The effects of UV and cycloheximide on SV40 amplification were superaddition. PMID- 2557543 TI - Detection and characterization of the fibroblast growth factor-related oncoprotein INT-2. AB - Products of the fibroblast growth factor-related proto-oncogene int-2 have been detected by using a monoclonal antibody and polyclonal antisera raised against synthetic peptides predicted from the DNA sequence. COS-1 monkey cells transfected with int-2 DNA linked to the simian virus 40 early promoter contained at least four int-2-specific proteins, presumably representing modified forms of the expected 27-kilodalton primary translation product. The level of expression was increased approximately six- to eightfold by mutation of sequences around the presumed initiation codon, negating their capacity to encode a short oligopeptide in the +1 reading frame. Both tunicamycin inhibition and in vitro translation experiments indicated that some of the modifications correspond to asparagine linked glycosylation, for which the sequence predicts a single site. In line with the similarities between INT-2 and other fibroblast growth factors, the in vitro translation products functioned as weak mitogens for mammary epithelial cells. PMID- 2557542 TI - Transcription from a murine T-cell receptor V beta promoter depends on a conserved decamer motif similar to the cyclic AMP response element. AB - We identified a regulatory region of the murine V beta promoter by both in vivo and in vitro analyses. The results of transient transfection assays indicated that the dominant transcription-activating element within the V beta 8.3 promoter is the palindromic motif identified previously as the conserved V beta decamer. Elimination of this element, by linear deletion or specific mutation, reduced transcriptional activity from this promoter by 10-fold. DNase I footprinting, gel mobility shift, and methylation interference assays confirmed that the palindrome acts as the binding site of a specific nuclear factor. In particular, the V beta promoter motif functioned in vitro as a high-affinity site for a previously characterized transcription activator, ATF. A consensus cyclic AMP response element (CRE) but not a consensus AP-1 site, can substitute for the decamer in vivo. These data suggest that cyclic AMP response element-binding protein (ATF/CREB) or related proteins activate V beta transcription. PMID- 2557545 TI - Rearrangement and diversification of immunoglobulin light-chain genes in lymphoid cells transformed by reticuloendotheliosis virus. AB - Avian lymphoid cells transformed by reticuloendotheliosis virus (REV-T) serve as a model to analyze the mechanism by which B-cell differentiation and antibody diversification occur in birds. Immunoglobulin light-chain gene rearrangements, diversification, and expression were analyzed in 72 independently derived REV-T transformed cell lines. Lymphoid cells transformed as the result of expression of the v-rel oncogene were divided into two distinct groups based on light-chain gene rearrangements. The status of the light-chain gene loci in these REV-T transformed cell lines was determined in part by the ages of the chickens whose spleen cells were transformed. In embryonic spleen cell lines transformed by the v-rel oncogene, rearrangements were not detected, even after prolonged culture in vitro, indicating that these cells are arrested in B-cell differentiation. REV-T transformants derived from spleens obtained from chickens 2 weeks old or older, however, had at least one light-chain allele rearranged. All of the cell lines analyzed which exhibited rearranged light-chain genes contained light-chain transcripts, and most of the REV-T-transformed cells which displayed light-chain rearrangements expressed immunoglobulin protein. REV-T, therefore, transforms B lymphoid cells at phenotypically different stages of development. Many REV-T transformed cells undergo immunoglobulin chain gene rearrangements during prolonged propagation in vitro. Most of the cell lines which rearrange their light-chain alleles also undergo diversification during cultivation in vitro. Light-chain diversification occurs during or after the rearrangement event. PMID- 2557544 TI - Tyrosine phosphorylation of a c-Src-like protein is increased in membranes of CD4 CD8- T lymphocytes from lpr/lpr mice. AB - Mice homozygous for the autosomal recessive lpr gene have a disorder that results in autoimmunity and massive accumulation of T lymphocytes lacking CD4 and CD8 surface markers. These abnormal T cells exhibit constitutive tyrosine phosphorylation of a component of the CD3-T-cell receptor complex. We compared membrane tyrosine phosphorylation in lpr/lpr CD4- CD8- T cells and control T cells, lpr membranes exhibited a 7.3-fold increase (n = 16) in tyrosine phosphorylation of a 60-kilodalton protein. The increase was correlated with the Lpr but not the CD4- CD8- phenotype in that p60 phosphorylation was not increased in membranes from normal CD4- CD8- thymocytes. To identify the p60 in lpr cells, we examined the activity of several T-cell tyrosine-specific protein kinases. p56lck phosphorylation was only slightly increased in lpr membranes (2.2-fold; n = 16). Phorbol ester treatment of intact T cells before membrane isolation caused p56lck to migrate as pp60lck; however, pp60lck could be clearly distinguished from the pp60 in lpr cells by two-dimensional gel electrophoresis. The pp60 from lpr cells exhibited several isoforms at pH approximately 6.3 to 6.5. Although on two-dimensional gels pp60c-src had a pI (6.4 to 6.8) within a similar region, p60c-src mRNA, protein, and kinase activities were not increased in lpr cells. In addition, staphylococcal V8 proteolytic cleavage of the lpr pp60 isolated on two dimensional gels yielded two major fragments, a pattern distinct from that of pp60c-src. However, by using an antiserum against the C-terminal sequence of c Src and other related kinases, including p59fyn, the pp60 could be immunoprecipitated in greater amounts from lpr than from control T cells. When pp59(fyn) was selectively immunoprecipitated from T-cell membranes with specific antisera, its molecular weight, proteolytic cleavage pattern, and behavior on two dimensional gels were identical to those of the pp60 from lpr cells. We conclude that p59(fyn) phosphorylation is increased in membranes from lpr/lpr CD4(-) CD8( ) T cells and that the increase is correlated with constitutive tyrosine phosphorylation and perhaps with the expansion of this unusual T-cell population. PMID- 2557546 TI - Mutational analysis of the Saccharomyces cerevisiae SNF1 protein kinase and evidence for functional interaction with the SNF4 protein. AB - The SNF1 gene of Saccharomyces cerevisiae encodes a protein-serine/threonine kinase that is required for derepression of gene expression in response to glucose limitation. We present evidence that the protein kinase activity is essential for SNF1 function: substitution of Arg for Lys in the putative ATP binding site results in a mutant phenotype. A polyhistidine tract near the N terminus was found to be dispensable. Deletion of the large region C terminal to the kinase domain only partially impaired SNF1 function, causing expression of invertase to be somewhat reduced but still glucose repressible. The function of the SNF4 gene, another component of the regulatory system, was required for maximal in vitro activity of the SNF1 protein kinase. Increased SNF1 gene dosage partially alleviated the requirement for SNF4. C-terminal deletions of SNF1 also reduced dependence on SNF4. Our findings suggest that SNF4 acts as a positive effector of the kinase but does not serve a regulatory function in signaling glucose availability. PMID- 2557548 TI - Molecular analyses of in vivo hprt mutations in human T-lymphocytes. III. Longitudinal study of hprt gene structural alterations and T-cell clonal origins. AB - The hprt clonal assay detects mutations occurring in vivo in the hypoxanthine guanine phosphoribosyltransferase (hprt) gene of human T-lymphocytes. Analysis of 94 wild-type and 326 hprt mutant clones from 3 normal males was performed using Southern blotting with hprt and T-cell receptor (TCR) gene probes. Gross structural alterations of the hprt gene occurred in approximately 14% of the in vivo derived mutants. Breakpoints were randomly distributed across the gene with one possible mutational "hot spot" observed. Most hprt mutants were independent as judged by TCR gene rearrangement patterns indicating that the measured hprt mutant frequency is a good measure of the actual hprt mutation frequency. However, sibling mutants (generally doublets and triplets except for one nonamer) were detected. Information on the timing in vivo of the hprt mutational events and the persistence in vivo of sibling mutants was also obtained. PMID- 2557549 TI - Mutations by near-ultraviolet radiation in Escherichia coli strains lacking superoxide dismutase. AB - In wild-type Escherichia coli, near-ultraviolet radiation (NUV) was only weakly mutagenic. However, in an allelic mutant strain (sodA sodB) that lacks both Mn- and Fe-superoxide dismutase (SOD) and assumed to have excess superoxide anion (O2 ), NUV induced a 9-fold increase in mutation above the level that normally occurs in this double mutant. When a sodA sodB double mutant contained a plasmid carrying katG+ (excess HP-I catalase), mutation by NUV was reduced to wild-type (sodA+ sodB+) levels. Also, in the sodA sodB xthA triple mutant, which lacks exonuclease III (exoIII) in addition to SOD, the mutational frequency by NUV was reduced to wild-type levels. This synergistic action of NUV and O2- suggested that pre-mutational lesions occur, with exoIII converting these lesions to stable mutants. Exposure to H2O2 induced a 2.8-fold increase in mutations in sodA sodB double mutants, but was reduced to control levels when a plasmid carrying katG+ was introduced. These results suggest that NUV, in addition to its other effects on cells, increases mutations indirectly by increasing the flux of OH. radicals, possibly by generating excess H2O2. PMID- 2557547 TI - UV-induced DNA damage is an intermediate step in UV-induced expression of human immunodeficiency virus type 1, collagenase, c-fos, and metallothionein. AB - UV irradiation of human and murine cells enhances the transcription of several genes. Here we report on the primary target of relevant UV absorption, on pathways leading to gene activation, and on the elements receiving the UV-induced signal in the human immunodeficiency virus type 1 (HIV-1) long terminal repeat, in the gene coding for collagenase, and in the cellular oncogene fos. In order to induce the expression of genes. UV radiation needs to be absorbed by DNA and to cause DNA damage of the kind that cannot be repaired by cells from patients with xeroderma pigmentosum group A. UV-induced activation of the three genes is mediated by the major enhancer elements (located between nucleotide positions 105 and -79 of HIV-1, between positions -72 and -65 of the collagenase gene, and between positions -320 and -299 of fos). These elements share no apparent sequence motif and bind different trans-acting proteins; a member of the NF kappa B family binds to the HIV-1 enhancer, the heterodimer of Jun and Fos (AP-1) binds to the collagenase enhancer, and the serum response factors p67 and p62 bind to fos. DNA-binding activities of the factors recognizing the HIV-1 and collagenase enhancers are augmented in extracts from UV-treated cells. The increase in activity is due to posttranslational modification. While AP-1 resides in the nucleus and must be modulated there, NF kappa B is activated in the cytoplasm, indicating the existence of a cytoplasmic signal transduction pathway triggered by UV-induced DNA damage. In addition to activation, new synthesis of AP-1 is induced by UV radiation. PMID- 2557551 TI - [The effect of acetylcholine and cholinergic transmission blockers on the neuronal impulse activity of the cat motor cortex in a conditioned reflex]. AB - Influence of acetylcholine, atropine, d-turbocurarine and hexonium on the background and evoked impulse activity of the motor cortex neurons was investigated in cat under instrumental placing reaction. Only those neurons were analyzed whose responses preceded conditional movement. It was shown that acetylcholine increased the background impulse activity in one group of cortical neurons and decreased it in the other neuronal group. During iontophoretic application of acetylcholine the background activity of some neurons declined in spite of the increase of their evoked conditional response. Atropine inhibited background impulse activity, the conditional impulse response being preserved. The background impulse activity increased and evoked conditional response was inhibited during iontophoretic application. Hexonium inhibited the background activity and conditional evoked impulse activity. It is suggested that cholinergic fibers in the natural state participate in supporting the background activity of cortical neurons by means of m-cholinoreceptors and at the sam time they evoke the impulse activity linked with conditional responses through n cholinoreceptors. PMID- 2557550 TI - Reactive oxygen-mediated damage to murine mammary tumor cells. AB - We have shown, in a preliminary report, that macrophages can induce strand breaks in the DNA of co-cultured tumor cells (Chong et al., 1988). The present study is designed to determine if oxygen-centered species generated by the cell-free enzyme-substrate combination of hypoxanthine and xanthine oxidase can induce similar lesions and to identify the specific mediator(s). We report that co incubation of murine mammary tumor cell lines with hypoxanthine and xanthine oxidase leads to the induction of DNA-strand breaks as determined by fluorescence analysis of DNA unwinding (FADU) assay or alkaline elution techniques. This damage is preventable by catalase which removes hydrogen peroxide but no protection is provided by agents to remove or prevent the formation of superoxide anion (superoxide dismutase), or hydroxyl radical (mannitol or the iron chelator o-phenanthroline). Likewise, cyclooxygenase or lipoxygenase inhibitors of arachidonate metabolism (indomethacin, nordihydroguaiaretic acid, caffeic acid) or bromophenacyl bromide do not alter the degree of DNA scission. Treatment with higher doses of oxygen species leads to significant toxicity as determined by evaluation of cell growth potential or colony-forming ability. Again, toxicity is prevented only by the presence of catalase. Tumor cells are able to rejoin strand breaks at lower, less toxic doses. When comparing different tumor cell subpopulations at various stages of progression, i.e., metastatic vs. nonmetastatic, for sensitivity to hydrogen peroxide-induced strand breakage, we found that at lower concentrations (less than 5 microM) metastatic populations are sensitive whereas nonmetastatic populations exhibit no significant breakage. At higher concentrations of hydrogen peroxide, all lines were sensitive, suggesting that a lower threshold of sensitivity may exist for more progressed tumor cell lines. PMID- 2557552 TI - [Numerical characteristics of the structure of the vesicular apparatus of the synapses in the dorsal horn of the cat spinal cord]. AB - Numerical parameters characterizing the structure of vesicular apparatus of presynaptic terminals were determined on the basis of the morphometrical investigation of the ultrastructure of 135 synapses from the dorsal horn of the cat spinal cord. Statistical estimates of the number of vesicles in the terminal was obtained (mean value 470). Bimodality of distribution of distances from a centre of each vesicle to the nearest site of the active zone has been demonstrated, that is regarded as a possible structural correlate of the two-pool model of the transmitter storage. The possibility of classification of synapses according to the spatial distribution of vesicles, relations between such distribution and parameters of the transmitter release and storage are discussed. PMID- 2557553 TI - [Excitation and inhibition processes in the neurons of the thalamic motor nuclei of normal cats and following an N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine induced lesion of the nigrostriatal dopaminergic system]. AB - Peculiarities of excitation and inhibition evoked in motor thalamic nuclei (VA VL) neurons by electrical stimulation of red nucleus were studied on intact cats and after injection of N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP; 5 mg/kg i.m., p.d. during five days). Two days after the last injection as much as 48% of nigral neurons were destroyed and the content of dopamine in the caudate nucleus fell to 30% as compared to intact animals. Before acute experiments all cats were anaesthetized with ketalar and immobilized with myorelaxine. It was found that MPTP injections caused a decrease of the inhibition duration and effectiveness in relay and nonrelay VA-VL neurons. The inhibition deficiency was accompanied by shortening of latencies of orthodromic responses evoked by red nucleus stimulation and facilitation of antidromic spikes invasion into somata of relay neurons after motor cortex stimulation. It was suggested that the reduction of GABAergic nigro-thalamic influences modulated by dopamine underlay the developing deficiency of inhibition. PMID- 2557554 TI - [Disordered associative long-term potentiation in the hippocampus after the tetanization of the reinforcing input]. AB - Field potentials (FPs) were recorded from stratum radiatum of CA1 region of mouse hippocampal slices after weak orthodromic stimulation (C2). Associative long-term potentiation (ALTP), induced by simultaneous tetanization of C2 with strong tetanization of another group of fibres (C1) was investigated. Effects of the additional tetanization of C1 delivered 50-300 ms before and 50-300 ms after conjoint tetanization of two inputs were compared. In experiments with intervals 50-200 ms preliminary tetanizations of C1 suppressed ALTP (the FPs amplitude increase was 10.4 +/- 5.2%) in comparison with procedure in which an additional tetanization of C1 follows conjoint tetanization (the FPs increase was 32.4 +/- 5.3%). There was no significant difference between two procedures in experiments with 300 ms intervals. Three mechanisms have been considered to explain the influence of the preliminary strong tetanus on ALTP: inactivation of "fast" calcium channels, afterburst hyperpolarization and synaptic inhibition. It is suggested that mechanisms of this suppression are similar to mechanisms underlying relative inefficacy of the "backward" association procedure in behavioral conditioning. PMID- 2557555 TI - A prospective study of a rapid method for diagnosing cytomegalovirus infections in immunosuppressed patients. AB - A rapid assay is described for detection of cytomegalovirus in peripheral blood lymphocytes. It consists of an indirect immunofluorescence technique for detection of cytomegalovirus antigens by means of a monoclonal antibody directed against early viral coded proteins. This assay was compared with the conventional cell culture system. Patients transplanted between December 1986 and May 1988 were studied, and of 27 patients identified, two were excluded due to early graft failure. A total of 320 blood specimens obtained were studied, and from 12 patients cytomegalovirus was isolated in at least one specimen by conventional cell culture (in total 25 specimens). Results were available with the new technique within 6 h, whereas the cell culture took an average of 13 days to develop the typical cytopathic effects changes. Sensitivity and specificity compared with that of viral isolation in conventional cell culture was 92% and 95%, respectively. This technique provides an accurate and rapid diagnosis of cytomegalovirus infections, and allows specific antiviral therapy to be started earlier. PMID- 2557556 TI - Involvement of the mu-opiate receptor in peripheral analgesia. AB - The intradermal injection of mu (morphine, Tyr-D-Ala-Gly-NMe-Phe-Gly-ol and morphiceptin), kappa (trans-3,4-dichloro-N-methyl-N[2-(1-pyrrolidinyl) cyclohexyl]benzeneactemide) and delta ([D-Pen2.5]-enkephalin and [D-Ser2] [Leu]enkephalin-Thr) selective opioid-agonists, by themselves, did not significantly affect the mechanical nociceptive threshold in the hindpaw of the rat. Intradermal injection of mu, but not delta or kappa opioid-agonists, however, produced dose-dependent inhibition of prostaglandin E2-induced hyperalgesia. The analgesic effect of the mu-agonist morphine was dose dependently antagonized by naloxone and prevented by co-injection of pertussis toxin. Morphine did not, however, alter the hyperalgesia induced by 8-bromo cyclic adenosine monophosphate. We conclude that the analgesic action of opioids on the peripheral terminals of primary afferents is via a binding site with characteristics of the mu-opioid receptor and that this action is mediated by inhibition of the cyclic adenosine monophosphate second messenger system. PMID- 2557557 TI - Mediation of primary afferent peripheral hyperalgesia by the cAMP second messenger system. AB - Cyclooxygenase (prostaglandin E2 and prostaglandin I2) and lipoxygenase [8(R), 15(S)-dihydroxyicosa-(5E-9,11,13Z)-tetraenoic acid] products of arachidonic acid metabolism are thought to produce peripheral hyperalgesia by a direct action on the primary afferent nociceptor. In this study we investigated the possibility that these eicosanoids generate hyperalgesia through a common second messenger in the rat. We report that 8-bromo cAMP, a membrane permeable analogue of cAMP, produces a dose-dependent hyperalgesia that is not affected by treatments that interrupt indirect routes of hyperalgesia production including sympathectomy with 6-hydroxydopamine, depletion of polymorphonuclear leukocytes (a source of hyperalgesic eicosanoids) with hydroxyurea, or blockade of the cyclooxygenase pathway of arachidonic acid metabolism with indomethacin. The phosphodiesterase inhibitor isobutyl-methylxanthine markedly prolongs the hyperalgesic effect of 8 bromo cAMP as well as those of the directly acting hyperalgesic agents prostaglandin E2, prostaglandin I2 and 8(R),15(S)-dihydroxyicosa-(5E-9,11,13Z) tetraenoic acid. We conclude that the effect of all known hyperalgesic eicosanoids is mediated by the cAMP second messenger system and suggest, therefore, that cAMP mediates peripheral hyperalgesia in primary afferent nociceptors. PMID- 2557558 TI - Excitatory amino acid receptors in the human cerebral cortex: a quantitative autoradiographic study comparing the distributions of [3H]TCP, [3H]glycine, L [3H]glutamate, [3H]AMPA and [3H]kainic acid binding sites. AB - The excitatory amino acids are probably the major neurotransmitters in the cerebral cortex, and they act through at least three receptors: the N-methyl-D aspartate, the quisqualate and the kainic acid receptors. Under the appropriate conditions, [3H]1-(1-(2-thienyl)-cyclohexyl)piperidine [( 3H]TCP), [3H]glycine and L-[3H]glutamate label different sites on the N-methyl-D-aspartate receptor, [3H]-alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid [( 3H]AMPA) labels the quisqualate receptor and [3H]kainic acid the kainic acid receptor. The anatomical localizations of these binding sites were studied in sections of blocks removed from the cerebral cortices of eight post-mortem human brains. The results showed that, in the human cerebral cortex, [3H]TCP, [3H]glycine and L [3H]glutamate binding sites had congruent distributions, with [3H]AMPA binding sites showing a similar distribution. In the hippocampus, these four ligands had high binding site densities in the CA1 region and the dentate gyrus molecular layer. With the exception of the striate cortex, in the neocortex, a tri-laminar pattern was seen consisting of a high density across laminae I-III, a layer of low density corresponding to the region of lamina IV, and a band of moderate density across laminae V and VI, except for [3H]AMPA where the middle zone of low density was usually wider. [3H]Kainic acid showed a binding pattern which was generally complementary to that of the other four ligands. There were low levels of [3H]kainic acid binding sites in the CA1 region of the hippocampus with higher levels in the CA3 region, the hilus, and the inner third of the dentate gyrus molecular layer. In the neocortex there was a band of high density corresponding to laminae V and VI, with a thin band of moderate binding corresponding to lamina I and the outer region of lamina II. An exception was the motor cortex where the highest level of [3H]kainic acid binding was in laminae I and II. The high degree of congruence between the binding patterns of [3H]TCP, [3H]glycine and L [3H]glutamate (using conditions appropriate for the N-methyl-D-aspartate receptor) supports data indicating that these ligands bind to different regions of the same receptor complex. The similar distribution of [3H]AMPA binding sites, with the exception of the striate cortex, supports observations made in rodents that N-methyl-D-aspartate receptors and quisqualate receptors have similar distributions and perform different but related functions in excitatory transmission.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557559 TI - Distribution of major neurotransmitter receptors in the motor and somatosensory cortex of the rhesus monkey. AB - The in vitro quantitative autoradiographic technique was used to characterize the distributions of alpha 1, alpha 2, beta 1 and beta 2 adrenergic, D1 and D2 dopaminergic, 5-HT1 and 5-HT2 serotonergic, M1 and M2 cholinergic, GABAA and benzodiazepine receptors in the motor (Brodmann's area 4) and somatosensory (Brodmann's areas 3, 1 and 2) cortex of the adult rhesus monkey. All receptor subtypes studied were present throughout all layers of both areas. In the somatosensory cortex, each receptor had its own laminar distribution. Some subtypes of the same receptor (5-HT1 and 5-HT2; alpha 1 and alpha 2) had complementary distributions while others (beta 1 and beta 2; D1 and D2; M1 and M2) had largely overlapping distributions. In contrast, different receptors had remarkably coincidental distributions in the motor cortex. In this area, they all tended to concentrate in layers I, II and the upper part of layer III. However, such coextensive distribution of many types of neurotransmitter receptors is not observed in motor cortex of rats and humans and therefore may be a distinctive feature of motor cortex in the rhesus monkey. The findings described in this paper indicate that somatosensory and motor areas are distinct in their receptor architecture and that receptor autoradiography provides a useful complement to classical histological techniques in elucidating areal differences in the cortex. PMID- 2557560 TI - On the properties and origin of the GABAB inhibitory postsynaptic potential recorded in morphologically identified projection cells of the cat dorsal lateral geniculate nucleus. AB - Intracellular recordings were performed from projection cells of the cat dorsal lateral geniculate nucleus in vitro to investigate the properties and origin of optic tract evoked inhibitory postsynaptic potentials mediated by GABAB receptors and their relationship to the physiologically different cell classes present in this nucleus. In all three main laminae of the dorsal lateral geniculate nucleus, stimulation of the optic tract evoked an excitatory postsynaptic potential followed by two inhibitory postsynaptic potentials. The first is a GABAA receptor mediated inhibitory postsynaptic potential since it was blocked by bicuculline, reversed in polarity following intracellular Cl- injection and had a reversal potential similar to the bicuculline sensitive hyperpolarizing effect of GABA. The second is a GABAB receptor mediated inhibitory postsynaptic potential. Its amplitude was not linearly related to membrane potential (maximal amplitude at 60 mV), it decreased when using frequencies of stimulation higher than 0.05 Hz and it was reversibly increased by addition of bicuculline to the perfusion medium. The reversal potential of GABAB inhibitory postsynaptic potentials was dependent on the extracellular K+ concentration but did not change in the presence of bicuculline or when recording with Cl- filled microelectrodes. While GABAA inhibitory postsynaptic potentials always abolished repetitive firing of projection cells, GABAB inhibitory postsynaptic potentials were able to block weak firing but unable to decrease strong activation of projection cells evoked by direct current injection. Optic tract evoked GABAB (as well as GABAA) inhibitory postsynaptic potentials could be recorded in slices which did not include the perigeniculate nucleus, thus indicating that they are generated by the interneurons of the dorsal lateral geniculate nucleus. Using intracellular injection of horseradish peroxidase, we have found that the GABAB inhibitory postsynaptic potentials are present in projection cells showing many different types of neuronal morphologies. In conclusion, GABA released from interneurons in the dorsal lateral geniculate nucleus is capable of evoking an early, short lasting GABAA and a late, long-lasting GABAB inhibitory postsynaptic potential in projection cells with diverse morphology, indicating that the late inhibition in the dorsal lateral geniculate nucleus can no longer be associated exclusively with the recurrent inhibitory pathway through the perigeniculate nucleus. PMID- 2557561 TI - Cytochemical restoration in the upper dorsal horn after transganglionic degenerative atrophy: temporospatial and fine structural correlates. AB - Reorganization of synaptic circuitry has been studied in the upper dorsal horn (Lamina II, substantia gelatinosa Rolandi) of the lumbar spinal cord in the adult rat, by means of electron histochemical visualization of thiamine monophosphatase after transganglionic degenerative atrophy. Thiamine monophosphatase, a highly specific and selective marker of Type C (small) dorsal root neurons, was demonstrated at light and electron microscopic levels by means of a Gomori-type cytochemical reaction, using thiamine monophosphate chloride (Sigma) as substrate and Pb2+ as a capturing agent. Transganglionic degenerative atrophy, induced by a crush injury of the sciatic nerve, results in partial depletion of thiamine monophosphatase from ipsilateral segments L2-S1. The extent of depletion was determined in a complete series of frozen cross-sections, by means of measuring the projections of active and depleted areas and their distances from the midline. Values were fed into a personal computer and maps demonstrating the distribution of intact and impaired areas were generated. The V-shaped area of depletion starts to shrink due to incipient regeneration on the 23rd postoperative day, in caudorostral and mediolateral gradients. Replenishment of thiamine monophosphatase is completed on the 60th postoperative day. Electron microscopic cytochemistry revealed the presence of the thiamine monophosphatase reaction end product in axonal growth cones, filopodia, young axons and their varicous swellings that are transformed into scallopped en passant terminals in the later course of regeneration. Axonal growth cones and regenerating sprouts undergo Wallerian degeneration and simultaneous redepletion of the marker enzyme after transection of dorsal roots L3, L4 and L5. Thiamine monophosphatase, located initially within the axoplasms of regenerating fibers, is successively translocated to the external axolemmal surfaces. Functional maturity of the terminals is achieved only later. The resulting redundant and transient wiring is thinned out in a following maturation period, in a manner resembling the sequence of events in embryonic development. The regenerative potency of central terminals of primary sensory neurons is not restricted to a single regeneration. By repeatedly crushing the sciatic nerve, four successive degenerative-regenerative cycles have been evoked, resulting in replenishment of the marker enzyme thiamine monophosphatase. PMID- 2557562 TI - Differential actions of the blockade of spinal opioid, adrenergic and serotonergic receptors on the tail-flick inhibition induced by morphine microinjected into dorsal raphe and central gray in rats. AB - Microinjection of morphine sulfate into dorsal raphe, ventrolateral central gray and dorsolateral central gray inhibits spinal nociceptive reflexes. The effects of the blockade of spinal opioid, adrenergic, and serotonergic receptors by intrathecal injection of naloxone, yohimbine and methysergide, respectively, on inhibition of the tail-flick response induced by morphine microinjected into dorsal raphe, ventrolateral central gray and dorsolateral central gray were studied. Naloxone (20 micrograms) given intrathecally effectively antagonized inhibition of the tail-flick response induced by morphine (4 micrograms) given into dorsal raphe and ventrolateral central gray, but not dorsolateral central gray. On the other hand, intrathecal injection of yohimbine (30 micrograms) antagonized inhibition of the tail-flick response induced by morphine given into ventrolateral central gray and dorsolateral central gray, but not dorsal raphe. Intrathecal injection of prazosin (30 micrograms) did not antagonize inhibition of the tail-flick response induced by morphine given into dorsal raphe or lateral central gray. Intrathecal injection of methysergide (30 micrograms) only partially antagonized inhibition of the tail-flick response induced by morphine given into dorsal raphe, but not ventrolateral central gray and dorsolateral central gray. It is concluded that the analgesia induced by morphine injected into dorsal raphe is mediated by spinal opioid receptors but not by spinal alpha 2-adrenergic receptors while the analgesia produced by morphine given into dorsolateral central gray is mediated by spinal alpha 2-adrenergic receptors. The analgesia induced by morphine given into ventrolateral central gray is mediated in part by both spinal alpha 2-adrenergic and opioid receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557563 TI - [Role of diet in the treatment of constipation. Review of the literature]. AB - Constipation represents a common social phenomenon today, correlated with the habits of diet and life. Many studies have shown the lack of alimentary fiber as a cause of constipation. In analyzing the different types, the characteristics, the physical and metabolical action of fiber, it has been shown that it is necessary to introduce fiber into the everyday diet to be able to prevent or cure constipation. In addition, it has been seen that there has been a reduction in the consumption of fiber in many nations and some reasons have been investigated. PMID- 2557564 TI - [Immunological aspects in anti-HIV seronegative drug addicts. Correlation with the duration of heroin addiction and other viral infections]. AB - In order to determine whether the immunological abnormalities described in intravenous drug addicts (IDA), are due to HIV infection, other viral infections or to the abuse of narcotic drugs, we studied the T lymphocyte subsets and serological markers of infection with hepatitis B and delta virus, cytomegalovirus and Epstein Barr virus, in 49 IDA. The immunological and serological features of IDA were compared with the control group, made up of 20 healthy subjects. In intravenous drug abusers we found a significant increase in the number of total lymphocytes (P less than 0.01), T-lymphocytes (P less than 0.05), T-suppressor cells (P less than 0.05), and serum IgG levels (P less than 0.0001) as compared with the control group. The prevalence of serological markers of infection with hepatitis B virus, hepatitis delta virus, cytomegalovirus and Epstein Barr virus was significantly higher in IDA as compared with the controls. In conclusion our study demonstrates that T-lymphocyte subsets in IDA seronegative for HIV infection are characterized by an enhancement of peripheral lymphocyte cells with a normal OKT4/OKT8 ratio. PMID- 2557565 TI - Extracellular adenosine 5'-triphosphate-evoked norepinephrine secretion not relating to voltage-gated Ca channels in pheochromocytoma PC12 cells. AB - Characteristics of extracellular ATP-stimulated norepinephrine (NE) secretion were investigated by the batch method in relation to membrane current measured with the whole cell voltage-clamp technique. Extracellular ATP stimulated [3H]NE secretion from PC12 cells in a concentration range from 10 microM to 1 mM. The maximal effect of ATP was about 5 times larger than that of nicotine. ATP stimulated secretion was extracellular Ca-dependent, but it was not inhibited by the Ca-channel blockers, cadmium (300 microM) or nicardipine (10 microM). Extracellular ATP activated an inward current in PC12 cells and the peak amplitude of the current was about 5 times larger than that activated by nicotine. These findings suggest that extracellular ATP activates receptor operated channels and causes NE secretion. PMID- 2557566 TI - Development of Na, K-ATPase in neocortical grafts. AB - Pieces of cerebral cortex from 14-day rat embryos were transplanted into freshly prepared cavities in the cerebral cortex of adult rats. At various time intervals after implantation. Na,K-ATPase, Mg-ATPase as well as the ratio of two molecular forms of Na,K-ATPase were determined in the grafts and compared with the values obtained from intact cortex at various stages of postnatal development. Up to the 57th postimplantation day (PID), Na.K-ATPase develops very slowly, reaching on PID 57, i.e. on postconceptional day (PCD) 71, only 40% of the enzyme activity of intact brain cortex (PND 49, PCD 71). At PID 90 (PCD 104) Na,K-ATPase activity attained that of the intact adult tissue (PND 82, PCD 104). Mg-ATPase activity in the grafts developed similarly to that of intact brain but was much higher in the early postimplantation phase. The maximum ratio of the two molecular forms of Na,K-ATPase [alfa(+)/alfa] was shifted toward later developmental periods in the grafts as compared with the intact cerebral cortex. PMID- 2557567 TI - Activation of protein kinase C by 4-aminopyridine dependent on Na+ channel activity in rat hippocampal slices. AB - The convulsant drug 4-aminopyridine (4-AP) stimulates the phosphorylation of the neuron-specific presynaptic protein B-50 in hippocampal slices. This effect could be attenuated by the protein kinase C (PKC) inhibitor staurosporine. Moreover, the endogenous phosphorylation of B-50 was found to be restricted to the 15 kDa Staphylococcus aureus protease fragment of B-50, known to contain the PKC acceptor site. The effect of 4-AP on B-50 phosphorylation was sensitive to the Na+ channel blocker tetrodotoxin. These results indicate that 4-AP stimulates PKC activity in hippocampal slices by a mechanism dependent on Na+ channel activity. PMID- 2557569 TI - Dietary regulation of the intestinal vitamin D receptor. AB - A low calcium diet resulted in significant upregulation of the concentration of the vitamin D receptor in the intestine of the rat. PMID- 2557568 TI - Ligand-binding characteristics of [3H]QNB, [3H]prazosin, [3H]rauwolscine, [3H]TCP and [3H]nitrendipine to cerebral cortical and hippocampal membranes of senescence accelerated mouse. AB - The senescence accelerated mouse (SAM) is known as a murine model of aging and memory dysfunction. In the cerebral cortical membranes of male 9-month-old SAM mice, the Bmax values of [3H]rauwolscine and [3H]nitrendipine binding, and the values of both Kd and Bmax of [3H]TCP binding in the accelerated aging strain SAM P/8, were significantly increased compared with the values in the control strain SAM-R/1. In hippocampal membranes, however, the Bmax values of [3H]quinuclidinyl benzilate and [3H]nitrendipine binding were significantly decreased in SAM-P/8 compared with those in SAM-R/1. These results suggest that muscarinic acetylcholine receptors, alpha 2-adrenoceptors, N-methyl-D-aspartate receptor channels and L-type Ca2+ channels are changed in cerebral cortex and hippocampus in SAM-P/8 at 9 months. PMID- 2557570 TI - Early effects of intra-arterial chemotherapy in patients with brain tumours studied with PET: preliminary results. AB - In ten patients with malignant gliomas the regional cerebral metabolic rate for glucose (rCMRGlc) was studied with positron emission tomography (PET) using 2-18F deoxyglucose (18FDG) before and within 1 to 7 days after intra-arterial chemotherapy with the nitrosourea derivative ACNU (iaACNU). Three patients were studied before and after two iaACNU courses and one patients before and after three iaACNU courses. The early effects of iaACNU on tumour rCMRGlc were highly variable and appeared to be more pronounced after the first course of iaACNU than in later iaACNU courses, i.e. more pronounced in untreated patients. Although there was no clear correlation between the change of rCMRGlc following the first course of iaACNU and the clinical outcome in this small group of patients, the patient with the most pronounced decrease of tumour metabolism (-16.5%) after the first course of iaACNU exhibited full tumour remission for 12 months, while the patient with the most pronounced increase of tumour metabolism (+65%) after the first course of iaACNU developed rapid tumour progression. The first results indicate that early effects of intra-arterial chemotherapy may be observed with 18FDG PET, especially following the first course of iACNU. Further studies are needed to evaluate the predictive value of such studies for therapy response. PMID- 2557571 TI - [Antimicrobial chemotherapy--its clinico-pharmacologic significance]. AB - Remembering of Markusovszky, the outstanding Hungarian physician of the last century, author quotes Markusovszky's comments on perspectives of infectious diseases, then outlines the development of antibacterial chemotherapy. Some problems are discussed concerning the clinical pharmacology of the aminoglycoside and cephalosporin antibiotics and those of the new quinolones as well. Some results of author's and her coworker's researches are presented. PMID- 2557572 TI - Low molecular weight heparin in the symptomatic treatment of chronic venous insufficiency. Controlled double blind study vs calcium heparin. AB - Ninety patients suffering from chronic venous insufficiency were submitted to subcutaneous treatment with calcium heparin in fixed doses (5000 I.U. t.i.d.) or with different doses of a low molecular weight heparin fraction 7,500 AXaU/day or 15,000 AXaU/day) for thirty days with a double blind random design. On days 0, 15 and 30, clinical parameters were assessed (edema, painfulness, cutaneous trophic lesions, paresthesia) and laboratory tests carried out (fibrinogenemia, plasma viscosity, euglobin lysis time, general tolerance parameters); at the beginning and at the end of the study the plethysmographic indexes of maximum venous flow and venous capacitance were evaluated. Treatment with 15,000 AXaU, of low molecular weight heparin caused modifications of the clinical and instrumental parameters which were on the whole better than those caused by heparin calcium t.i.d. throughout the study. The administration of 7,500 AXaU of low molecular weight heparin was practically equivalent to the control treatment at the end of the study, although at the intermediate assessment it was slightly less effective despite the fact that it had produced significant modifications with respect to the basal values. Treatment with low molecular weight heparin, with both doses used, produced fewer side effects than the reference drug. PMID- 2557573 TI - Liver transplantation for hepatoblastoma in a child with congenital absence of the portal vein. AB - Congenital absence of the portal vein with systemic visceral venous return in a 8 year-old girl with oculoauriculovertebral dysplasia (Goldenhar Syndrome) had been previously reported following its discovery during preoperative evaluation of a liver mass which was diagnosed as focal nodular hyperplasia after open biopsy. Subsequently, an enlarging diffuse hepatic neoplasm developed with associated elevated alpha feto-protein levels. Repeat biopsy and imaging showed the tumor to be a hepatoblastoma involving both lobes of the liver. The patient was treated by hepatic resection and orthotopic liver transplantation and is doing well at 18 months follow-up. PMID- 2557574 TI - Melanotic neuroectodermal tumor of infancy. MR findings and a review of the literature. AB - Melanotic neuroectodermal tumor of infancy is an uncommon neoplasm occurring primarily in the child one year or less in age. Difficulty in deciding the cellular origin of this tumor has led to numerous names, including congenital melanocarcinoma, melanotic epithelial odontoma, melanotic ameloblastoma, and retinal anlage tumor, to list a few. Electron microscopy and histochemical studies, however, have now established the neural crest as the most likely origin. The most frequent site of occurrence is the maxilla followed by the skull, the brain and the mandible. The genital organs are the most frequent extracranial site. Within the skull, there is a predilection for the anterior fontanel. The following is a case report of a young child with melanotic neuroectodermal tumor of infancy arising at the anterior fontanel. Included is a discussion of magnetic resonance (MR) findings, which to our knowledge, have not been previously reported in this tumor. PMID- 2557575 TI - Nucleotide sequence of a full length cDNA clone encoding a polyubiquitin gene from Pisum sativum. PMID- 2557576 TI - Cloning and sequencing of the polymerase gene (P) of Sendai virus (strain Fushimi). PMID- 2557577 TI - Cloning and sequencing of the polymerase gene (P) of Sendai virus (strain 6/94). PMID- 2557579 TI - The complete nucleic acid sequence of gene segment 3 of the IDIR strain of group B rotavirus. PMID- 2557578 TI - Nucleotide sequence of the gene encoding subunit 3 of cytochrome c oxidase (cox3) in the mitochondrial genome of Schizosaccharomyces pombe strain EF1. PMID- 2557581 TI - A simple and rapid method for the isolation of plasmid and lambda phage DNAs. PMID- 2557580 TI - Cloning and nucleotide sequence of the mouse Na,K-ATPase beta-subunit. PMID- 2557582 TI - A new series of CAT expression vectors. PMID- 2557583 TI - Screening and characterization of restriction endonucleases from a bacterial culture collection in Hong Kong. PMID- 2557584 TI - Rapid method for introducing restriction sites into double stranded plasmid DNA. PMID- 2557585 TI - Hemophilia B in a male with a four-base insertion that arose in the germline of his mother. PMID- 2557587 TI - The presence of the region on pBR322 that encodes resistance to tetracycline is responsible for high levels of plasmid DNA knotting in Escherichia coli DNA topoisomerase I deletion mutant. AB - Plasmid pBR322 DNA isolated from Escherichia coli DNA topoisomerase I deletion mutant DM800 is estimated to contain about 10% of the knotted forms (Shishido et al., 1987). These knotted DNA species were shown to have the same primary structure as usual, unknotted pBR322 DNA. Analysis of the knotting level of deletion, insertion and sequence-rearranged derivatives of pBR322 in DM800 showed that the presence of the region on pBR322 encoding resistance to tetracycline (tet) is required for high levels of plasmid knotting. When the entire tet region is present in a native orientation, the level of knotting is highest. Inactivating the tet promoter is manifested by a middle level of knotting. For deletion derivatives lacking various portions of the tet region, the level of knotting ranges from lowest to high depending on the site and length of the tet gene remaining. Inverting the orientation of tet region on the pBR322 genome results in a middle level of knotting. Deleting the ampicillin-resistance (bla)gene outside of its second promoter does not affect the level of knotting, if the entire tet gene remains. A possible mechanism of regulation of plasmid knotting is discussed. PMID- 2557586 TI - Evolutionary relationship between luteoviruses and other RNA plant viruses based on sequence motifs in their putative RNA polymerases and nucleic acid helicases. AB - Comparative studies of sequence motifs in the RNA polymerases and nucleic acid helicases of positive-sense RNA plant viruses have provided a new scheme for the classification of these pathogens. We propose a new luteovirus supergroup which should be added to the already described Sindbisvirus-like and picornavirus-like supergroups. Sequence motifs of nucleic acid helicases and RNA polymerases which previously were considered to be specific for each of the two supergroups now occur together within this new supergroup. We propose that this new viral supergroup provides an evolutionary link between the other two supergroups. PMID- 2557588 TI - Cytosine methylation in CTF and Sp1 recognition sites of an HSV tk promoter: effects on transcription in vivo and on factor binding in vitro. AB - We methylated specific cytosine residues within or immediately around the CTF and Sp1 binding sites of the Herpes simplex virus thymidine kinase promoter. The efficiency of transcription in vivo was reduced at least 50-fold compared with transcription from the unmethylated promoter. However, methylation within the CTF recognition site had no effect on the affinity of CTF for this site in vitro. Methylation of the Sp1 site resulted in only a small decrease in the affinity of this factor for its recognition site. In vivo studies showed that the same gene inserted in different vector DNAs was regulated differently by methylation in the promoter. These results show that cytosine methylation can inhibit transcription by a mechanism other than directly blocking the binding of transcription factors. PMID- 2557589 TI - Consensus DNA site for the Escherichia coli catabolite gene activator protein (CAP): CAP exhibits a 450-fold higher affinity for the consensus DNA site than for the E. coli lac DNA site. AB - We have synthesized two 40 base pair DNA fragments; one fragment contains the consensus DNA site for CAP (fragment 'ICAP'); the other fragment contains the E. coli lac promoter DNA site for CAP (fragment 'LCAP'). We have investigated the binding of CAP to the two DNA fragments using the nitrocellulose filter binding assay. Under standard conditions [( NaCl] = 200 mM, pH = 7.3), CAP exhibits a 450 fold higher affinity for ICAP than for LCAP. The salt dependence of the binding equilibrium indicates that CAP makes eight ion pairs with ICAP, but only six ion pairs with LCAP. Approximately half of the difference in binding free energy for interaction of CAP with ICAP vs. LCAP is attributable to this difference in ion pair formation. The pH dependence of the binding equilibrium indicates that the eight CAP-ICAP ion pairs and the six CAP-LCAP ion pairs do not involve His residues of CAP. PMID- 2557590 TI - Superhelical stress restrained in plasmid DNA during repair synthesis initiated by the UvrA, B and C proteins in vitro. AB - Purified UvrA, UvrB, UvrC, UvrD, PolA and Lig proteins from Escherichia coli have been used to assess the effect of nucleotide excision repair on the conformation of native negatively supercoiled plasmid DNA in an in vitro test system. The analysis of labeled reaction products on specific gel systems suggests that the Uvr excinuclease has the ability to restrain the superhelical stress in the template DNA during the repair process. This feature, observed in the case of the Uvr system is not found if the repair reaction is initiated by T4 endonuclease V or Micrococcus luteus UV endonuclease. PMID- 2557592 TI - Nucleotide and deduced amino acid sequence of the nonstructural protein, NS1, of the US bluetongue virus serotype 17. PMID- 2557591 TI - Hyperexpression and purification of Escherichia coli adenylate cyclase using a vector designed for expression of lethal gene products. AB - We describe the construction of a new generation of vectors (pRE) for the hyperexpression of lethal gene products such as adenylate cyclase in Escherichia coli. The pRE vectors are based on the lambda PL promoter and lambda cII ribosome binding site described by Shimatake and Rosenberg (Nature, 292, 128-132, 1981). They have a unique NdeI restriction endonuclease site 3' of the lambda cII ribosome binding site that includes the ATG initiation codon, multilinker cloning sites 3' to the NdeI site, and two lambda transcription terminators 5' and 3' of the lambda PL promoter to eliminate nonspecific transcription and reduce leaky PL transcription, respectively. For hyperexpression of adenylate cyclase, tight control of transcription was necessary since elevation of cAMP levels above the physiological range is lethal to E. coli. Lethality associated with the overproduction of adenylate cyclase was shown to be mediated through the cAMP receptor protein. We used this expression system to overproduce adenylate cyclase 7500 fold, corresponding to 30% of the total cellular protein. Under these conditions the enzyme precipitated with significant loss of activity. Reducing the rate and amount of adenylate cyclase expression to 16% of the total cell protein produced one fourth of the enzyme in a soluble form with high specific activity. The soluble adenylate cyclase was purified to near homogeneity. PMID- 2557593 TI - A screen for SVgpt in E. coli DH5 delta lac and DH5 alpha: small colony phenotype. PMID- 2557594 TI - Hsp82-neo transposition vectors to study insertional mutagenesis in Drosophila melanogaster and tissue culture cells. PMID- 2557595 TI - Four-base-pair deletion polymorphism at D5S71 (C11p11) linked to APC in the human chromosome 5q21-q22 region. PMID- 2557596 TI - Mixed germ cell tumour of the mediastinum (seminoma, embryonal carcinoma, choriocarcinoma and teratoma). Light and electron microscopic cytology and histological investigation. AB - A mixed germ cell tumour of the mediastinum was found in a 27-year-old subject. It had four components: seminoma, embryonal carcinoma, choriocarcinoma and teratoma. Material for light microscopy (L.M.) and ultrastructural studies were obtained with computerized tomography (CT) guided fine needle aspiration. Three histotypes were identified with ultrastructural studies while only seminoma was identified with L.M. The slides prepared for L.M. were reexamined in the light of findings with electron microscopy (E.M.) and previously unidentified cellular elements were found to be characteristic of choriocarcinoma and teratoma. Histological examination of surgical and autopsy material confirmed the cytological findings. Because of the singularity of this case, the rarity of this type of neoplasm and the difficulty in arriving at a differential diagnosis from other neoplastic and non neoplastic lesions, only the ultrastructural cytological examination allowed us to identify the mixed nature of the tumour. Such an identification is not always possible with L.M. but it is a determining factor in the prognosis and therapy. PMID- 2557597 TI - Multivisceral intestinal transplantation: surgical pathology. AB - We report the diagnostic surgical pathology of two children who underwent multivisceral abdominal transplantation and survived for 1 month and 6 months. There is little relevant literature, and diagnostic criteria for the various clinical possibilities are not established; this is made more complicated by the simultaneous occurrence of more than one process. We based our interpretations on conventional histology, augmented with immunohistology, including HLA staining that distinguished graft from host cells in situ. In some instances functional analysis of T cells propagated from the same biopsies was available and was used to corroborate morphological interpretations. A wide spectrum of changes was encountered. Graft-versus-host disease, a prime concern before surgery, was not seen. Rejection was severe in 1 patient, not present in the other, and both had evidence of lymphoproliferative disease, which was related to Epstein-Barr virus. Bacterial translocation through the gut wall was also a feature in both children. This paper documents and illustrates the various diagnostic possibilities. PMID- 2557598 TI - Fine needle aspiration cytology in advanced pediatric tumors. AB - Eighty-one children with clinically suspected malignant tumors were subjected to percutaneous fine needle aspiration cytology (FNAC) at the Pathology Department of the National Institute of Child Health, Jinnah Postgraduate Medical Centre, Karachi, from August 1986 through July 1987. There were 47 malignant diagnoses including lymphoma, neuroblastoma, nephroblastoma, Ewing's sarcoma, and leukemia. Histological findings confirmed the FNAC diagnoses in 36 cases in which a subsequent incisional biopsy or surgically removed specimen was available. FNAC results were confirmed in all benign cases. In 10 advanced cases of NonHodgkin's lymphoma, surgery was not possible because of marked malnourishment. One false negative and no false positive result was encountered. Forty-eight were females and thirty-three males. FNAC can be a quick, effective, and inexpensive alternative to open biopsy, particularly in advanced cases of malignancy in undernourished children where anesthesia and immediate surgery are contraindicated. PMID- 2557599 TI - Herpes simplex tracheobronchitis and pneumonitis. PMID- 2557600 TI - Thoracic disc herniation in acromegaly. AB - Herniation of a thoracic disc in an acromegalic giant is reported. Degenerative changes in the spine in association with dorsal kyphosis, and the additional strain, resulted in the disc prolapse. PMID- 2557602 TI - Mammography and the primary care management of breast lumps. AB - Breast cancer is the commonest cause of death from malignancy in women in the Western world. Although advances have been made in treatment, 11,000 women a year continue to die from the disease in Britain. Prognosis is undoubtedly better with early diagnosis, and this may best be achieved by prompt referral to a breast unit if a persistent mass is suspected clinically. PMID- 2557603 TI - Linear and cyclic alpha-melanotropin [4-10]-fragment analogues that exhibit superpotency and residual activity. AB - Two analogues of alpha-MSH (Ac-Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Lys-Pro Val-NH2), Ac-[Nle4, Asp5, D-Phe7, Lys10]alpha-MSH4-10NH2 and Ac-[Nle4, Asp5, D Phe7, Lys10] alpha-MSH4-10-NH2, were synthesized, and the melanotropic activities of the peptides were compared in several bioassays. Potencies were determined in the in vitro frog and lizard skin bioassays and in the S91 melanoma cell tyrosinase assay. Both analogues were equipotent or more potent than alpha-MSH in all bioassays, and the activities of the analogues were prolonged compared to alpha-MSH. The two analogues were very resistant to inactivation by purified proteolytic enzymes (alpha-chymotrypsin, trypsin, and pepsin). The two peptides could be topically applied and transdermally delivered across the skin of mice in vivo, resulting in a shift from pheomelanogenesis to eumelanogenesis within follicular melanocytes. The cyclic analogue exhibited greater potency, prolonged activity, and stability against enzyme inactivation than did the linear peptide. The significance of the findings for the further design of melanotropin analogues is discussed, as in the possible relevance of these melanotropin analogues for use in biomedical studies. PMID- 2557604 TI - Neural control of motile activity of light-sensitive iridophores in the neon tetra. AB - Experiments with skin pieces revealed that the sympathetic nervous system controls the activity of the light-sensitive iridophores in the stripes of the neon tetra. The spectral peak reflected from the cells was shifted toward longer wavelengths as a result of a direct interaction between norepinephrine and alpha adrenoceptors present on the cell membrane. Adenosine accelerated the recovery from the effects of the amine. Such regulation seems to operate when fish are in an excited state or under stress. Since alpha-melanophore-stimulating hormone (alpha-MSH), melanin-concentrating hormone (MCH) and melatonin caused the responses only at high concentrations, it is possible that these peptides and amine do not affect the properties of the light-reflecting cells in vivo. PMID- 2557605 TI - Absence of murine melanoma 5'-nucleotidase activity is not attributable to a detectable inhibitor. AB - Previous observations indicated the absence of demonstrable 5'-nucleotidase activity in six of seven cultured murine melanoma cell lines. It could not be determined from those studies whether the enzyme was absent, or whether an inhibitor was present. The current studies indicate that no inhibitor can be demonstrated, therefore the enzyme is absent. PMID- 2557606 TI - Use of an alpha-melanocyte-stimulating hormone analogue to improve alpha melanocyte-stimulating hormone receptor binding assay in human melanoma. AB - In order to optimize the detection and measurement of alpha-melanocyte stimulating hormone (alpha-MSH) receptivity in human melanoma cells, and the authors replaced the natural hormone by [Nle4,D-Phe7]-alpha-MSH, a more stable and potent analogue in the receptor binding assay commonly performed with alpha MSH. The following parameters were investigated: temperature, incubation time, number of cells, and ratio of labelled to unlabelled hormone. Optimal conditions for each assay were determined. The results demonstrate that the analogue has identical binding sites to alpha-MSH, as similar reciprocal displacements of each labelled (125I) hormone by serial dilutions of unlabelled alpha-MSH or [Nle4,D Phe7]-alpha-MSH (10(-12) M to 10(-6) M) were obtained. To further compare the two hormones, we performed a screening of various human cell lines: ten melanomas and five nonmelanomas. The assay with [Nle4,D-Phe7]-alpha-MSH yielded more receptor expression on six of ten melanoma lines against only four of ten with the natural hormone. In conclusion, the use of radiolabelled [Nle4,D-Phe7]-alpha-MSH analogue instead of labelled alpha-MSH improved both sensitivity and reproducibility in this receptor binding assay on human melanoma lines. PMID- 2557601 TI - Airway receptors. PMID- 2557607 TI - Alpha-melanocyte-stimulating hormone immunoreactivity in human melanoma metastases extracts. AB - Seven human melanoma metastases were extracted in order to check the possible presence of any alpha-melanocyte stimulating hormone (MSH) immunoreactivity. The aim of that study was to provide some explanation for, mainly, two observations that we have already made and reported: 1) increased plasma alpha-MSH levels in melanoma of tumour-bearing patients as compared with tumour-free patients; 2) the presence of specific alpha-MSH receptors on human melanoma cells in culture. We could measure large amounts of immunoreactive alpha-MSH in all tumours ranging from 0.31 to 4.27 pmoles per g of wet tissue. Further identification of the extracted material by high-performance liquid chromatography revealed compounds of higher molecular weight and more hydrophobic than synthetic alpha-MSH. In addition, purified extracts could also displace 125I-labelled alpha-MSH from its cellular binding sites in an alpha-MSH specific radio-receptor binding assay. Our findings would suggest a possible presence of some hormone precursor(s) inside the melanoma tumours. PMID- 2557608 TI - Expression of virally transduced mouse tyrosinase in tyrosinase-negative chick embryo melanocytes in culture. AB - A cDNA encoding mouse tyrosinase was inserted into a plasmid containing the provirus of a replication competent avian leukosis virus (ALV). A viral stock produced from the plasmid was used to infect cultured tyrosinase-negative (ca/ca) unpigmented chick embryo melanocytes. Five days after infection many cells were producing very dark discrete melanosomes. PMID- 2557610 TI - [Cushing's syndrome in unilateral diffuse-nodular hyperplasia of the adrenal cortex with a corticosteroma and in combination with a pheochromocytoma]. PMID- 2557609 TI - [Identification and characteristics of the beta-adrenergic receptors in the membranes of cultured endothelial cells of the human pulmonary artery]. AB - The properties of beta-adrenoreceptors (beta-AR) have been studied in endothelial cells from the human pulmonary artery (EPA) and umbilical vein (EUV). [125I] Iodosyanopindolol binding assay revealed the amount of 22 and 12 fmol/10(6) cells as well as Kd = 92 and 52 pM for EPA and EUV, respectively. Adrenergic agonists increased the cAMP levels in EPA in the order of potency characteristic for beta 2-AR: isoproterenol greater than epinephrine greater than norepinephrine. Basing on the results obtained in the present study and the literature data a conclusion is made that the properties of beta-AR in vascular endothelial cells do not practically depend on their localization, suggesting universality of their function within the vascular system. PMID- 2557611 TI - [Age-related morphofunctional characteristics of the adrenal cortical substance when ACTH is administered]. AB - The structural and morphometric features of the adrenocortical substance (ACS) were investigated in adult (5 mos.) and aged (24 mos.) albino male rats during single and prolonged ACTH administration (Synachten-Depot, Ciba). In single administration of a large dose of ACTH (50 micrograms per 100 g of body mass) to adult rats hypertrophy of ACS secretory cells was marked to a greater extent (by 105.9%), and the area of ACS cell nuclei was increased after the administration of a smaller dose of ACTH (25 micrograms per 100 g of body mass). In old animals the administration of a smaller dose of the hormone resulted in an increased area of nuclei and cytoplasm of ACS cells (by 55%). In prolonged administration of the hormone during 14 days an increase in the sizes of nuclei and cytoplasm of ACS cells was observed both in adult and aged rats. However in the adrenal glands of old rats, cell hypertrophy was less marked than that in adult animals (134 and 186.2%, respectively) and was accompanied by destructive changes in a portion of cells. In 95% the results of morphometry were obtained in the confidence interval using a Leitz-ASM unit (FRG) for semiautomatic image analysis. The differences were statistically significant. PMID- 2557612 TI - The Huntington disease locus is most likely within 325 kilobases of the chromosome 4p telomere. AB - The genetic defect responsible for Huntington disease was originally localized near the tip of the short arm of chromosome 4 by genetic linkage to the locus D4S10. Several markers closer to Huntington disease have since been isolated, but these all appear to be proximal to the defect. A physical map that extends from the most distal of these loci, D4S90, to the telomere of chromosome 4 was constructed. This map identifies at least two CpG islands as markers for Huntington disease candidate genes and places the most likely location of the Huntington disease defect remarkably close (within 325 kilobases) to the telomere. PMID- 2557613 TI - CpG island clones from a deletion encompassing the gene for adenomatous polyposis coli. AB - Adenomatous polyposis coli (APC), a dominantly inherited disorder, has been mapped to chromosome 5q15-q21 by family linkage studies. Cells from patients with deletions in this region, in one case associated with polyposis in a family, have been used to construct human hamster hybrid cell lines that retain either the normal or deleted chromosome 5. These lines have been used to identify markers from the region of the polyposis gene obtained by cloning the ends of 0.5- to 2 megabase BssHII fragments purified by pulsed-field gel electrophoresis. Three markers are described that map within the deletions and must therefore be close to the APC gene. PMID- 2557614 TI - Epstein-Barr virus/complement fragment C3d receptor (CR2) reacts with p53, a cellular antioncogene-encoded membrane phosphoprotein: detection by polyclonal anti-idiotypic anti-CR2 antibodies. AB - Epstein-Barr virus and the C3d fragment of the third component of complement are specific extracellular ligands for complement receptor type 2 (CR2). However, intracellular proteins that react specifically with CR2 and are involved in post membrane signals remain unknown. We recently prepared polyclonal anti-idiotypic anti-CR2 antibodies (Ab2) by using the highly purified CR2 molecule as original immunogen. We showed that Ab2 contained anti-idiotypic specificities that mimicked extracellular domains of CR2 and detected two distinct binding sites on CR2 for its specific extracellular ligands, Epstein-Barr virus and C3d. We postulated that Ab2 might also contain specificities that could mimic intracellular domains of CR2. Here we report that Ab2, which did not react with Raji B-lymphoma cell surface components, detected specifically, among all components solubilized from Raji cell membranes, a single intracellular membrane protein of apparent molecular mass of 53 kDa. This protein was identified as the p53 cellular antioncogene-encoded membrane phosphoprotein by analyzing its antigenic properties with Pab1801, a monoclonal anti-p53 antibody, and by comparing its biochemical properties with those of p53. Additionally, solubilized and purified CR2 bound to solubilized p53 immobilized on Pab1801-Sepharose. p53, like CR2, was localized only in purified plasma membranes and nuclei of Raji cells. These data suggest strongly that p53, a cellular antioncogene-encoded phosphoprotein, reacted specifically with CR2 in Raji membranes. This interaction may represent one of the important steps through which CR2 could be involved in human B-lymphocyte proliferation and transformation. PMID- 2557615 TI - Nerve growth factor induces growth and differentiation of human B lymphocytes. AB - Nerve growth factor (NGF) is known to affect peripheral sympathetic and sensory neurons as well as defined populations of neurons in the central nervous system. This paper presents evidence that NGF is also active in modulation of B-cell mediated immune responses. NGF receptors were immunoprecipitated from highly purified human B-cell populations, and to a lesser extent, from T-cell populations, by using a monoclonal antibody recognizing NGF receptors present on neural cells. NGF receptors were also detected in significant amounts in human spleen and lymph node tissue. In addition, NGF induced a dose-dependent increase in B-cell DNA synthesis as determined by incorporation of [3H]thymidine. This B cell growth-promoting activity was inhibited by a neutralizing anti-NGF monoclonal antibody. Immunoglobulin secretion, principally affecting IgM synthesis, was also modulated by NGF. The concentrations that affected B-cell proliferation are consistent with the presence of functional high-affinity NGF receptors. The results suggest that NGF, in addition to its neurotrophic function, also acts as an immunoregulatory cytokine. PMID- 2557616 TI - Cooperation of c-raf-1 and c-myc protooncogenes in the neoplastic transformation of simian virus 40 large tumor antigen-immortalized human bronchial epithelial cells. AB - Overexpression of c-raf-1 and the myc family of protooncogenes is primarily associated with small cell carcinoma, which accounts for approximately 25% of human lung cancer. To determine the functional significance of the c-raf-1 and/or c-myc gene expression in lung carcinogenesis and to delineate the relationship between protooncogene expression and tumor phenotype, we introduced both protooncogenes, alone or in combination, into human bronchial epithelial cells. Two retroviral recombinants, pZip-raf and pZip-myc, containing the complete coding sequences of the human c-raf-1 and murine c-myc genes, respectively, were constructed and transfected into simian virus 40 large tumor antigen-immortalized bronchial epithelial cells (BEAS-2B); this was followed by selection for G418 resistance. BEAS-2B cells expressing both the transfected c-raf-1 and c-myc sequences formed large cell carcinomas in athymic nude mice with a latency of 4 21 weeks, whereas either pZip-raf- or pZip-myc-transfected cells were nontumorigenic after 12 months. Cell lines established from tumors (designated RMT) revealed the presence of the cotransfected c-raf-1 and c-myc sequences and expressed morphological, chromosomal, and isoenzyme markers, which identified BEAS-2B cells as the progenitor line of the tumors. A significant increase in the mRNA levels of neuron-specific enolase was detected in BEAS-2B cells containing both the c-raf-1 and c-myc genes and derived tumor cell lines. The data demonstrate that the concomitant expression of the c-raf and c-myc protooncogenes causes neoplastic transformation of human bronchial epithelial cells resulting in large cell carcinomas with certain neuroendocrine markers. The presented model system should be useful in studies of molecular events involved in multistage lung carcinogenesis. PMID- 2557618 TI - Identification of cell surface receptors for the 86-kilodalton glycoprotein of human cytomegalovirus. AB - Cell surface receptors for the 86-kDa glycoprotein (gp86) of human cytomegalovirus (HCMV) were identified by using two monoclonal anti-idiotype antibodies that bear the internal image of gp86. These antibodies bound to cells permissive for HCMV infection by both ELISA and immunofluorescence assay and inhibited HCMV plaque formation in human embryonic lung (HEL) cells. Immunoblot analysis showed specific binding of both internal image anti-idiotype antibodies as well as gp86 to an HEL cell membrane protein with an approximate molecular mass of 92.5 kDa. In addition, immunoprecipitation of radiolabeled membrane and cell surface proteins from human foreskin tissue, human foreskin fibroblasts, or HEL cells showed specific binding of anti-idiotype antibody predominantly to the 92.5-kDa protein. PMID- 2557617 TI - Positive inotropic effects of the endogenous Na+/K(+)-transporting ATPase inhibitor from the hypothalamus. AB - Bovine hypothalamus contains a nonpeptidic substance that inhibits purified Na+/K(+)-transporting ATPase [ATP phosphohydrolase (Na+/K(+)-transporting), EC 3.6.1.37] reversibly with high affinity by a mechanism similar to, but not identical to, that of the cardiac glycosides. It possesses some of the characteristics ascribed to a putative endogenous "digitalis-like" compound that has been implicated in the control of renal sodium excretion and the pathogenesis of essential hypertension in man. To determine whether this hypothalamic Na+/K(+) transporting ATPase inhibitor might have physiologic properties in cardiac tissues, its effects on Na+ pump inhibition, accumulation of cytosolic free calcium, and contractile response were studied in cultured, spontaneously contracting neonatal rat cardiocytes. The hypothalamic factor potently inhibited the Na+ pump in these cells, increased myoplasmic free calcium in a dose dependent manner, and reversibly enhanced myocyte contractility by up to 40%, comparable in degree to maximal positive inotropic effects caused by the cardiac glycoside ouabain. Comparative studies further indicate that cardiotoxic effects of ouabain in the myocytes may be more complex than simple progressive elevation of intracellular free calcium concentration because at a free calcium concentration in excess of that produced by a toxic dose of ouabain, no toxicity with the hypothalamic Na+/K(+)-transporting ATPase inhibitor occurred. PMID- 2557619 TI - v-src induces clonal sarcomas and rapid metastasis following transduction with a replication-defective retrovirus. AB - v-src is an effective carcinogen when expressed from Rous sarcoma virus (RSV) in vivo. Whereas RSV tumors require sustained oncogene expression, their growth is largely a balance between viral recruitment of tissues and host immune destruction of infected cells. We have therefore examined the tumorigenic potential of v-src in the absence of viral recruitment and viral antigen expression. v-src was introduced with high efficiency into chicken wing web tissues using replication-defective (rd) retroviral vectors. Clonal sarcomas were induced rapidly, and, furthermore, v-src potentiated metastatic progression in approximately 0.1%-1% of tumor clones with unexpectedly short latency. rd vectors proved effective not only in transducing v-src into tissues but also as insertional markers of tumor clonality. The rd vector present in most primary and metastatic tumors was a highly truncated form of RSV derived by viral transmission of spliced v-src mRNA; this vector should thus avoid viral recruitment and host anti-viral immune reaction through its complete lack of viral structural genes. Under such conditions v-src maintains strong carcinogenicity in vivo when restricted to clonal tumor growth and can confer rapid metastatic potential on a discrete subset of tumor clones. PMID- 2557620 TI - Gamma-aminobutyrate type B receptor modulation of L-type calcium channel current at bipolar cell terminals in the retina of the tiger salamander. AB - Bipolar-cell axon terminals receive direct synaptic input from amacrine-cell processes, suggesting a possible pathway for modulation of transmitter release. In retinal slices, bath-applied baclofen, a gamma-aminobutyrate type B (GABAB) receptor agonist, reduced a patch-clamp-recorded L-type calcium channel current in a population of bipolar cells with axon terminals that ramify along the midline of the inner plexiform layer. Lucifer yellow staining revealed that this current was found only in bipolar cells that retain axon terminals and their associated telodendria, suggesting that the current is generated at the terminal and also possibly modulated there. T-type calcium currents were found in all bipolar cells, including those without axon terminals, but were not modulated by baclofen. The baclofen-induced reduction of calcium current was enhanced by guanosine 5'-[gamma-thio]triphosphate and eliminated by guanosine 5'-[beta thio]diphosphate added to the cytoplasm by the patch recording electrode, suggesting that the GABAB receptors act through a guanine nucleotide-binding regulatory protein (G protein). Baclofen also reduced an excitatory synaptic input to a population of amacrine cells with processes that ramify along the midline of the inner plexiform layer--cells probably postsynaptic to the bipolar terminals. This suggests that GABAB receptors modulate not only the calcium current but also transmitter release by a pathway involving G proteins and L-type calcium channels. PMID- 2557621 TI - Differential expression of three gap junction proteins in developing and mature brain tissues. AB - By using antibodies directed against gap junction proteins of liver (connexins 26 and 32) and heart (connexin 43), we have localized immunoreactivity to specific cell types in frozen sections of adult rodent brains. Connexin 32 reactivity was found in oligodendrocytes and also in a few neurons, whereas reactivity to connexins 26 and 43 was localized to leptomeningeal cells, ependymal cells, and pineal gland. Immunoreactivity with antibodies to connexin 43 also occurred in astrocytes. Furthermore, during embryonic and postnatal maturation of brain tissues, gap junction proteins were differentially expressed. Connexins 43 and 26 predominated in the neuroepithelium of embryonic brains, whereas connexin 32 was virtually absent. Between 3 and 6 weeks after birth, connexin 26 largely disappeared from immature brain; this time course corresponded to the increased expression of connexin 32. Expression of connexin 43 remained high throughout embryonic and postnatal development. These findings demonstrate that gap junction expression in the brain is diverse, with specific cell types expressing different connexins; this cell-specific distribution may imply differences in the function of these intercellular channels in different loci and developmental stages. PMID- 2557622 TI - Localization of the receptor site for alpha-scorpion toxins by antibody mapping: implications for sodium channel topology. AB - Site-directed and monoclonal antibodies recognizing different extracellular regions of the RII sodium channel alpha subunit have been used to determine the sequences that comprise the receptor for alpha-scorpion toxins by evaluating the effect of antibody on voltage-dependent binding of radio-labeled toxin isolated from Leiurus quinquestriatus to both reconstituted rat brain sodium channel and rat brain synaptosomes. Of six antibodies tested, two recognizing amino acid residues 355-371 and 382-400 located on an extracellular loop between transmembrane segments S5 and S6 of domain I and one recognizing residues 1686 1703 of a similar loop of domain IV inhibit binding by 30-55%. Inhibition is concentration-(EC50 = 0.4-2 microM) and time- (t1/2 = 40-80 min) dependent. Five different monoclonal antibodies recognizing the same extracellular loop in domain I inhibit binding completely with similar EC50 values as observed for site directed antibodies. Kinetic studies of the antibody effect are consistent with a slowly reversible competition for the toxin receptor site. Our results suggest that the extracellular loops between segments S5 and S6 of domains I and IV comprise at least part of the alpha-scorpion toxin receptor site and support the membrane topology models in which domains I and IV are adjacent in the tertiary structure of the channel protein and six transmembrane sequences are contained in each of the four homologous domains. PMID- 2557623 TI - Evolution of a bifunctional enzyme: 6-phosphofructo-2-kinase/fructose-2,6 bisphosphatase. AB - The bifunctional rat liver enzyme 6-phosphofructo-2-kinase/fructose-2,6 bisphosphatase (ATP:D-fructose-6-phosphate 2-phosphotransferase/D-fructose-2,6 bisphosphate 2-phosphohydrolase, EC 2.7.1.105/EC 3.1.3.46) is constructed of two independent catalytic domains. We present evidence that the kinase and bisphosphatase halves of the bifunctional enzyme are, respectively, structurally similar to the glycolytic enzymes 6-phosphofructo-1-kinase and phosphoglycerate mutase. Computer-assisted modeling of the C-terminal bisphosphatase domain reveals a hydrophobic core and active site residue constellation equivalent to the yeast mutase structure; structural differences map to length-variable, surface-located loops. Sequence patterns derived from the structural alignment of mutases and the bisphosphatase further detect a significant similarity to a family of acid phosphatases. The N-terminal kinase domain, in turn, is predicted to form a nucleotide-binding fold that is analogous to a segment of 6 phosphofructo-1-kinase, suggesting that these unrelated enzymes bind fructose 6 phosphate and ATP substrates in a similar geometry. This analysis indicates that the bifunctional enzyme is the likely product of gene fusion of kinase and mutase/phosphatase catalytic units. PMID- 2557624 TI - Sequence-specific recognition and cleavage of duplex DNA via triple-helix formation by oligonucleotides covalently linked to a phenanthroline-copper chelate. AB - Homopyrimidine oligodeoxynucleotides recognize the major groove of the DNA double helix at homopurine.homopyrimidine sequences by forming local triple helices. Phenanthroline was covalently attached to the 5' end of an 11-mer homopyrimidine oligonucleotide of sequence d(TTTCCTCCTCT). Simian virus 40 DNA, which contains a single target site for this oligonucleotide, was used as a substrate for the phenanthroline-oligonucleotide conjugate. In the presence of copper ions and a reducing agent, a single specific double-strand cleavage site was observed at 20 degrees C by agarose gel electrophoresis. The efficiency of double-strand cleavage was greater than 70% at 20 degrees C and pH 7.4. Secondary cleavage sites were observed when binding of the oligonucleotide to mismatched sequences was allowed to take place at low temperature. The exact location of the cleavage sites was determined by polyacrylamide gel electrophoresis of denatured fragments by using both simian virus 40 DNA and a synthetic DNA fragment containing the target sequence. The asymmetric distribution of the cleavage sites on the two strands revealed that the cleavage reaction took place in the minor groove even though the phenanthroline linker was located in the major groove. Linkers of different lengths were used to tether phenanthroline to the oligonucleotide and their relative efficacies of DNA cleavage were compared. Based on these comparative studies and on model building, it is proposed that the phenanthroline ring carried by the oligonucleotide intercalates from the major groove and that copper chelation locks the complex in place from within the minor groove where the cleavage reaction occurs. PMID- 2557625 TI - Isolation of a cDNA clone of the 14-kDa subunit of the signal recognition particle by cross-hybridization of differently primed polymerase chain reactions. AB - Using an enhancement of the polymerase chain reaction (PCR) technique, we have isolated a complementary DNA encoding SRP14 (14-kDa subunit), one of six proteins contained in the signal recognition particle (SRP). Several pools of degenerate oligonucleotides encoding different peptide sequences of SRP14 were used to generate amplified DNA by the PCR. A cross-hybridization procedure was developed to identify the authentic SRP14 cDNA clone among the amplified DNA products obtained by PCR. The basis of this approach is the assumption that a partial cDNA of SRP14 should be the only DNA product common to two amplification reactions primed with different degenerate oligonucleotide mixtures. The partial canine cDNA of SRP14 identified by this procedure served as a probe to isolate a complete cDNA clone of SRP14 from a mouse embryonic cDNA library in lambda phage gt10. PMID- 2557626 TI - Multiple functions of human single-stranded-DNA binding protein in simian virus 40 DNA replication: single-strand stabilization and stimulation of DNA polymerases alpha and delta. AB - The human single-stranded-DNA binding protein (human SSB) is required for simian virus 40 (SV40) DNA replication in vitro. SV40 large tumor antigen and human SSB can support extensive unwinding of SV40 origin-containing DNA in the presence of ATP and a topoisomerase that relieves positive superhelicity. Although SSBs from viral and prokaryotic sources substituted for human SSB in the DNA-unwinding reaction, they did not substitute in the replication of SV40 DNA. The specificity for human SSB in SV40 DNA replication can be explained, at least in part, by the finding that DNA polymerase alpha was stimulated 10-fold by human SSB but not by other SSBs. Human SSB also stimulated proliferating-cell nuclear antigen dependent DNA polymerase delta; however, other SSBs stimulated this polymerase as well. PMID- 2557627 TI - An ochre mutation in the vitamin D receptor gene causes hereditary 1,25 dihydroxyvitamin D3-resistant rickets in three families. AB - Hereditary 1,25-dihydroxyvitamin D3-resistant rickets is a rare autosomal recessive disease resulting from target-organ resistance to the action of the active hormonal form of vitamin D. Four affected children from three related families with the classical syndrome of hereditary 1,25-dihydroxyvitamin D3 resistant rickets and the absence of detectable binding to the vitamin D receptor (VDR) in cultured fibroblasts or lymphoblasts were examined for genetic abnormalities in the VDR gene. Genomic DNA from Epstein-Barr virus-transformed lymphoblasts of eight family members was isolated and amplified by polymerase chain reaction techniques. Amplified fragments containing the eight structural exons encoding the VDR protein were sequenced. The DNA from all affected children exhibited a single C----A base substitution within exon 7 at nucleotide 970 that resulted in the conversion of the normal codon for tyrosine (TAC) into a premature termination codon (TAA) at amino acid 292. This mutation causes a truncation of the VDR protein thereby deleting a large portion of the steroid hormone binding domain (amino acids 292-424). Although the affected children were all homozygotic for the mutation, the four parents tested all exhibited both wild type and mutant alleles, indicating a heterozygous state. The functional consequences of this mutation were confirmed after expression of the recreated mutant VDR cDNA in mammalian cells. Recreated mutant receptor exhibited no specific 1,25-[3H]dihydroxyvitamin D3 binding and failed to activate a cotransfected VDR promoter-reporter gene construct. Thus these findings identify an ochre mutation in a human steroid hormone receptor in patients with hereditary 1,25-dihydroxyvitamin D3-resistant rickets. PMID- 2557628 TI - Primary structures of the heterogeneous nuclear ribonucleoprotein A2, B1, and C2 proteins: a diversity of RNA binding proteins is generated by small peptide inserts. AB - We have isolated cDNAs for the major heterogeneous nuclear ribonucleoprotein (hnRNP) A2, B1, and C2 proteins and determined their nucleotide and deduced amino acid sequences. The A2 and B1 cDNAs are identical except for a 36-nucleotide in frame insert in B1. Similarly, the sequence of the C2 protein cDNA is related to that of C1 in that C2 contains an extra 39 in-frame nucleotides. Therefore, the B1 amino acid sequence is identical to A2 except for the insertion of 12 amino acids near its amino terminus, and C1 and C2 are also identical to each other except for an extra 13 amino acids near the middle of C2. All three proteins are members of a large family of RNA binding proteins that contain the consensus sequence-type RNA binding domain (CS-RBD). The A2 and B1 proteins have a modular structure similar to that of the hnRNP protein A1: they contain two CS-RBDs and a glycine-rich auxiliary domain at the carboxyl terminus. The CS-RBDs of A2 and B1 have approximately 80% amino acid identity with those of A1, whereas the glycine rich auxiliary domain is considerably more divergent with less than 30% of the amino acids being identical. These findings indicate that the addition of small peptides, probably by alternative pre-mRNA splicing, generates some of the diversity apparent among hnRNP proteins. PMID- 2557629 TI - Mapping the active-site tyrosine of vaccinia virus DNA topoisomerase I. AB - Site-directed mutagenesis of the vaccinia virus gene encoding a type I DNA topoisomerase implicates Tyr-274 as the active-site residue that forms a covalent adduct with DNA during cycles of DNA-strand breakage and reunion. Replacement of Tyr-274 by phenylalanine results in loss of the ability of the enzyme to relax negatively supercoiled DNA as well as to form the covalent DNA-protein intermediate. Substitution of phenylalanine for tyrosine at nine other sites in the protein has no apparent effect on enzyme activity. Amino acid sequence alignment reveals Tyr-274 to be homologous to Tyr-727 and Tyr-771, respectively, of the type I topoisomerases from Saccharomyces cerevisiae and Saccharomyces pombe; Tyr-727 and Tyr-771 have been shown to represent the active-site tyrosines of those enzymes. Sequence comparison of the active-site regions defines a motif Ser-Lys-Xaa-Xaa-Tyr common to the viral and cellular type I topoisomerases, including the human enzyme. PMID- 2557630 TI - Binding of the herpes simplex virus major regulatory protein to viral DNA. AB - Infected-cell protein 4 (ICP4), the major regulatory protein specified by herpes simplex virus 1 in infected cells, binds to homologs of the sequence ATCGTCnnnnYCGRC (A sites, where n is any nucleotide, Y is a pyrimidine, and R is a purine) and to unrelated sequences for which no consensus sequence has been derived (B sites). We have examined the binding of ICP4 to each of two A and two B binding sites by using Fab fragments of a monoclonal antibody that is reactive with an epitope located at the N terminus of ICP4 and that decreases the mobility of ICP4-DNA complexes in non-denaturing gels. The results indicate that each type of site binds two monomers of ICP4. Methylation-interference studies on the type B sites mapped the guanines whose methylation interfered with the binding of ICP4. The methylation-interference pattern obtained with one of the B sites was similar to that obtained on an A site but differed from that of the other B site. The ability of ICP4 to bind to DNA fragments containing the binding site appears to be dependent on length and on the proximity of the binding site to the fragment end. Short DNA fragments did not form stable complexes with ICP4 even though they contained all of the purines whose methylation interfered with the binding of the regulatory protein. PMID- 2557631 TI - Mossbauer study of the inactive Fe3S4 and Fe3Se4 and the active Fe4Se4 forms of beef heart aconitase. AB - Active beef heart aconitase contains an iron-sulfur cluster with an [Fe4S4]2+ core. This cluster can be converted into Fe3S4 with concomitant loss of enzymatic activity. We have reconstituted apo-aconitase with iron and selenide to obtain Fe4Se4 aconitase. The Se analog has higher catalytic activity than the native S containing enzyme when isocitrate is the substrate. Oxidation of [Fe4Se4]2+ with ferricyanide yields the inactive [Fe3Se4]1+ form. The Se-containing 3-Fe cluster can be reduced to [Fe3Se4]0. We have studied the [Fe3S4]1+,0, [Fe3Se4]1+,0, and [Fe4Se4]2+ states with Mossbauer spectroscopy from 1.3 K to 200 K in magnetic fields up to 6.0 T. The spectra of the S- and Se-containing enzymes were found to be remarkably similar. The spectra of the 3-Fe clusters were analyzed and the salient features of the electronic structure are discussed. PMID- 2557632 TI - In vivo 23Na and 31P NMR measurement of a tonoplast Na+/H+ exchange process and its characteristics in two barley cultivars. AB - A Na+ uptake-associated vacuolar alkalinization was observed in roots of two barley cultivars (Arivat and the more salt-tolerant California Mariout) by using 23Na and 31P in vivo NMR spectroscopy. A NaCl uptake-associated broadening was also noted for both vacuolar Pi and intracellular Na NMR peaks, consistent with Na+ uptake into the same compartment as the vacuolar Pi. A close coupling of Na+ with H+ transport (presumably the Na+/H+ antiport) in vivo was evidenced by qualitative and quantitative correlations between Na+ accumulation and vacuolar alkalinization for both cultivars. Prolongation of the low NaCl pretreatment (30 mM) increased the activity of the putative antiport in Arivat but reduced it in California Mariout. This putative antiport also showed a dependence on NaCl concentration for California Mariout but not for Arivat. No cytoplasmic acidification accompanied the antiporter activity for either cultivar. The response of adenosine phosphates indicated that ATP utilization exceeded the capacity for ATP synthesis in Arivat, but the two processes seemed balanced in California Mariout. These comparisons provide clues to the role of the tonoplast Na+/H+ antiport and compensatory cytoplasmic adjustments including pH, osmolytes, and energy phosphates in governing the different salt tolerance of the two cultivars. PMID- 2557633 TI - A ubiquitin carrier protein from wheat germ is structurally and functionally similar to the yeast DNA repair enzyme encoded by RAD6. AB - The RAD6 gene from the yeast Saccharomyces cerevisiae encodes a ubiquitin carrier protein (E2) required for a variety of cellular processes including DNA repair, induced mutagenesis, and sporulation. Here we identify an E2 from a higher plant, wheat, that is similar to RAD6 with respect to both structure and in vitro substrate specificity. The protein was purified from wheat germ by a combination of ubiquitin covalent affinity chromatography and anion-exchange HPLC and has an apparent molecular mass of 23 kDa [referred to as E2(23 kDa)]. E2(23 kDa) was capable of binding ubiquitin by means of a thiol ester linkage in an ATP dependent and ubiquitin-activating enzyme-dependent reaction. In the presence of a variety of target proteins, E2(23 kDa), like the RAD6 gene product, formed covalent ubiquitin-protein conjugates in vitro only with histones in a ubiquitin protein ligase-independent reaction. E2(23 kDa) recognized both core and linker histones with an apparent order of preference of H2A greater than or equal to H1 greater than H2B greater than H3 greater than H4. This E2 protein was approximately 17-fold more effective at conjugating ubiquitin to histones than three other purified wheat germ E2 proteins tested. Mouse anti-E2(23 kDa) antibodies were used to isolate E2(23 kDa) DNA sequences from a wheat cDNA expression library. Antibody-positive clones were confirmed by amino acid identity of the sequence deduced from the cDNA to the peptide sequence of an E2(23 kDa) tryptic fragment. Protein expressed in Escherichia coli by the E2(23 kDa) cDNA was capable of both thiol ester adduct formation and conjugation of ubiquitin to histones. Analysis of the E2(23 kDa) cDNA shows that it encodes a protein with considerable amino acid sequence similarity to the yeast RAD6 gene product. Similarities exist at the amino terminus, the region surrounding the putative ubiquitin binding site, and at the carboxyl terminus, which is unusually acidic. Based on both the structural and enzymatic similarities to the RAD6 gene product, E2(23 kDa) may represent the first DNA repair enzyme identified in higher plants. PMID- 2557634 TI - The human integrin VLA-2 is a collagen receptor on some cells and a collagen/laminin receptor on others. AB - The integrin heterodimer VLA-2, previously known as a collagen receptor, is now shown also to be a laminin receptor. Adhesion of the human melanoma cell line LOX to laminin was inhibited by anti-VLA alpha 2 antibodies. Because VLA-2-mediated LOX cell attachment to laminin was not inhibited by digestion with collagenase, collagen contamination of laminin was not a factor. In addition, VLA-2 from LOX cells bound to immobilized laminin, and binding was disrupted by EDTA but not by Arg-Gly-Asp (RGD) peptides. VLA-3 also bound to laminin-Sepharose, although less avidly than VLA-2. Thus, at least four separate members of the integrin beta 1 subfamily serve as laminin receptors--i.e., VLA-2 and VLA-3 (this study) together with VLA-1 and VLA-6 (other reports). Whereas LOX and other cell lines used VLA-2 as both a laminin and collagen receptor, fibroblast VLA-2 mediated collagen but not laminin binding. Likewise, VLA-2 from platelets did not interact with laminin. Despite this functional discordancy, VLA-2 from laminin-binding and nonbinding sources was indistinguishable by all immunochemical and biochemical criteria examined. Thus, functional differences in VLA-2 may be due to cell type specific modulation. PMID- 2557636 TI - Multiple actions of THA on cholinergic neurotransmission in Alzheimer brains. AB - 1,2,3,4-tetrahydro-9-aminoacridine (THA) is a cholinesterase inhibitor presently under investigation in clinical trials for treatment of Alzheimer's disease, senile dementia of Alzheimer type (AD/SDAT). To further analyse the underlying mechanisms for its effect in human brain, an in vitro model which allows measurement of acetylcholine (ACh) release from human postmortem brain slices has been used. In control cortical tissue THA induces a decreased release of ACh probably due to negative feedback mechanisms mediated via presynaptic muscarinic autoreceptors. In AD/SDAT cortex THA enhances the release of ACh to control level. This effect is prevented by nicotinic or muscarinic receptor antagonists, which suggest receptor mechanisms involving both nicotinic and muscarinic receptors. Subchronic treatment of rats with THA (10 mg/kg sc twice daily) or physostigmine (0.9 mg/kg sc five times daily) causes a significant increase in the number of high affinity nicotinic receptors in the cortex of THA treated rats whereas no change is found in the physostigmine treated rats. The number of muscarinic receptors are decreased following both THA and physostigmine treatment. PMID- 2557637 TI - Alterations in adrenergic receptors of frontal cortex and cerebral microvessels in Alzheimer's disease and aging. AB - Biochemical and pathological abnormalities are evident in the noradrenergic innervation of the cerebral cortex in Alzheimer's Disease (AD), and there is also a decline in aging, which may lead to changes in adrenergic receptors. To assess this question, we analyzed adrenergic receptor subtypes by ligand binding methods in prefrontal cortex and brain microvessels from subjects with AD and aging controls. Ligand binding to adrenoceptors and their subtypes did not change with postmortem delay in obtaining tissues. alpha 1-adrenergic receptors of the frontal cortex did not correlate with age but there was a small (approximately 25%), though significant, reduction in AD subjects. In cerebral microvessels, there were no changes in these receptors. alpha 2-receptors of the cortex significantly declined with age in controls and were also significantly reduced by approximately 50% in AD subjects. However, in cerebral microvessels alpha 2 adrenergic receptors were significantly increased by approximately 60% in AD. We suggest that presynaptic alpha 2-adrenoceptors on noradrenergic synapses may be those that are selectively decreased in the prefrontal cortex in AD. Total beta receptors in cortex did not correlate with age, nor were they altered in AD. However, beta 1-receptors were decreased but beta 2-receptors were significantly increased in AD, indicating a change in the relative ratio of beta 1/beta 2 receptors. Similarly, beta 2-receptors of cerebral microvessels were significantly increased in AD. These changes suggest receptor "up-regulation" in response to noradrenergic denervation in AD and may reflect functional changes at the blood-brain barrier. PMID- 2557635 TI - Induction of the transcription factor IRF-1 and interferon-beta mRNAs by cytokines and activators of second-messenger pathways. AB - Nuclear protein IRF-1 (interferon regulatory factor 1) was earlier shown to bind to cis-acting regulatory elements present on interferon (IFN)-alpha/beta genes and some IFN-inducible genes. Here we show that in both human FS-4 and murine L929 cells, steady-state levels of IRF-1 mRNA were increased by treatment with tumor necrosis factor (TNF), interleukin 1 (IL-1), poly(I).poly(C), or IFN-beta. IRF-1 mRNA induction was also demonstrated in cells treated with calcium ionophore A23187 or with phorbol 12-myristate 13-acetate, but not with epidermal growth factor, dibutyryl-cAMP, or the adenylate cyclase activator forskolin. To determine whether stimulation of IRF-1 mRNA levels correlates with IFN-beta induction, we compared IRF-1 and IFN-beta mRNA levels in cells exposed to various stimuli. In L929 cells, treatment with poly(I).poly(C) under conditions that failed to induce significant levels of IFN-beta mRNA led to a very low induction of IRF-1 mRNA, but "priming" cells with IFN prior to the addition of poly(I).poly(C) greatly increased both IRF-1 and IFN-beta mRNAs. In FS-4 cells an increase in IFN-beta mRNA (examined by the polymerase chain reaction) was seen after treatment with TNF, IL-1, A23187, or poly(I).poly(C), but not with IFN beta, epidermal growth factor, dibutyryl-cAMP, or forskolin. Thus, all treatments that increased steady-state levels of IFN-beta mRNA also enhanced IRF-1 mRNA levels. However, treatment with IFN-beta, which caused a marked stimulation in IRF-1 mRNA, failed to produce a detectable increase in IFN-beta mRNA. It appears that IRF-1 may be necessary but not sufficient for IFN-beta induction. The ability of TNF and IL-1 to increase both IRF-1 and IFN-beta mRNAs may be responsible for some similarities in the actions of TNF, IL-1, and the IFNs. PMID- 2557638 TI - Nerve growth factor receptor expressing human basal forebrain neurons: pathologic alterations in Alzheimer's and Parkinson's disease. AB - Normal aged human basal forebrain tissue immunohistochemically stained with a monoclonal antibody for nerve growth factor receptor (NGFR) revealed a continuum of NGFR containing neurons in brain regions corresponding to Ch1-Ch4. NGFR immunoreactive neurons were observed within the medial septum (Ch1), vertical (Ch2) and horizontal (Ch3) limb nuclei of the diagonal band and nucleus basalis (Ch4) complex. Occasional dystrophic NGFR containing neurons were observed within the Ch subfields. Colocalization experiments demonstrated that virtually all (95%) NGFR containing perikarya stained positively for the specific cholinergic marker acetylcholinesterase. On the other hand, occasional cholinergic negative NGFR positive neurons were seen in the putamen of the striatal complex. In Alzheimer's disease, subfields of the basal forebrain revealed extensive neuronal loss and shrinkage, dystrophic fibers as well as normal appearing neurons. Each AD case exhibited an individual pattern of cell loss. Within the Ch1-4 subfields NGFR and ChAT colocalized, indicating that the NGFR remained coupled to cholinergic neurons in AD. In some Parkinson's cases there was a severe loss of NGFR containing basal forebrain neurons. These findings indicate that NGFR containing basal forebrain neurons colocalize with ChAT, are severely affected in AD, in some PD cases, and NGFR remains coupled to cholinergic perikarya in both neurodegenerative disorders. PMID- 2557640 TI - Serum protein interactions with neurotransmitter receptors: implications for Alzheimer's disease. PMID- 2557639 TI - Reduced cAMP-signal transduction in postmortem hippocampus of demented old people. AB - The basal as well as the stimulated activity of the adenylate cyclase was determined in postmortem hippocampi. The tissue probes were obtained from 12 demented individuals (10 Alzheimer-type dementia; 1 Down's syndrome; 1 argyrophilic grains syndrome) and from 15 age-matched controls. The diagnoses were done in accordance with histopathological criteria. Adenylate cyclase was stimulated by isoprenaline, Gpp(NH)p, or forskolin. The amount of cAMP formed was determined by the protein binding method using a radioimmuno assay. In tissues of controls as well as of demented patients adenylate cyclase was stimulated in the rank order of isoprenaline less than Gpp (NH) p less than forskolin. In hippocampal tissues of demented individuals a significant reduction (50%, p less than 0.01) in basal as well as stimulated adenylate cyclase activity was found. This reduction in cAMP signal transduction is not caused by simple cell loss. PMID- 2557641 TI - Ubiquitin: a multifunctional regulatory protein associated with the cytoskeleton. AB - The discovery that ubiquitin is associated with the neurofibrillary tangles of Alzheimer's disease and inclusion bodies of other neurodegenerative disorders raises the possibility that ubiquitin is playing a role in the pathology. Ubiquitin is a small, highly conserved protein that is found both free and covalently attached to other proteins in all eukaryotic cells. Ubiquitin is conjugated with some proteins that are rapidly degraded and with a select set of regulatory proteins that are relatively stable, implying that ubiquitin is multifunctional. We have produced monoclonal antibodies to ubiquitin and identified the epitopes. Different monoclonal antibodies recognize different ubiquitin-protein conjugates; this could be due to different conformational states of ubiquitin or to steric interference by the conjugated protein. These observations suggest that ubiquitin is structurally and therefore functionally different in different contexts. Ubiquitin is localized in several compartments in cells and this may also affect function. We discovered that it is a normal component of the microtubule network and have identified ubiquitinated proteins in bovine brain that appear associated with microtubules. We discovered that ubiquitin has intrinsic proteolytic activity and proposed that ubiquitin-protein conjugates could function as ad hoc proteases. This proteolytic activity offers a heuristic basis from which to explore ubiquitin's cellular functions. The association of ubiquitin with the cytoskeletal networks suggests that proteolytic processing is a function of these structures. Disruption of these structures in neurodegenerative disorders, improper localization of ubiquitinated components or appearance of abnormal conjugates could lead to altered proteolytic processing pathways and contribute to the pathology of these diseases. PMID- 2557642 TI - Intermediate filaments and ubiquitin: a new thread in the understanding of chronic neurodegenerative diseases. AB - We have recently shown that there is a previously unsuspected link between the intracellular inclusions seen in several major chronic human degenerative diseases, including neurodegenerative diseases: the inclusions showing ubiquitin immunoreactivity. The conditions include Parkinson's disease, motor neurone disease, Alzheimer's disease, Pick's disease, and alcoholic liver disease as well as cerebellar astrocytomas and a myopathy. The inclusions found in these diseases are reported to contain intermediate filaments: neurofilaments are associated with Lewy bodies in Parkinson's disease, Pick's bodies in Pick's disease and neurofibrillary tangles in Alzheimer's disease, cytokeratins are found in Mallory bodies in alcoholic liver disease, glial fibrillary acidic proteins and vimentin are found in Rosenthal fibres in astrocytomas, and desmin is found in cytoplasmic bodies in cytoplasmic body myopathy. Therefore five classes of intermediate filaments are found in inclusions which also contain ubiquitin immunoreactivity; we have also shown that ubiquitin immunoreactivity is present in vesicles in some areas of granulovacuolar degeneration in Alzheimer's disease. Protein ubiquitination is considered a signal for extralysosomal protein degradation, (although ubiquitination may have several other important functions). We have recently shown that intermediate filaments are involved in protein sequestration before degradation by lysosomally mediated autophagy: therefore intermediate filament-containing ubiquitinated inclusions may be the hallmarks of cellular attempts to eliminate pathogenic insults by the activation of both extralysosomal and lysosomal mechanisms of intracellular protein degradation. We have recently been able to reproduce, at least in part, some of the clinical observations in tissue culture cells. Ubiquitinated protein conjugates accumulate in lysosomes in fibroblasts treated with the lysosomal cysteine protease inhibitor E-64, which may mimic aspects of granulovacuolar degeneration. PMID- 2557644 TI - Tau and ubiquitin immunoreactivity at different stages of formation of Alzheimer neurofibrillary tangles. AB - In his original 1911 publication Alois Alzheimer classified neurofibrillary tangles (ANT) into three morphologically defined subgroups according to their stage of maturation. The present study shows that changes in the morphological appearance of ANT during their maturation process are accompanied by changes in their antigenic profile. As shown by several immunocytochemical studies these abnormal phosphorylated microtubule-associated protein tau and of ubiquitin. In this study, immunoreactivity for the altered tau is not only seen in a subset of tangles but also in the cytoplasm of some nerve cells lacking ANT, which we believe to be at a stage of neuronal alteration preceding the formation of compact tangles (Stage 0 tangles). Similar numbers of Stage 0 tangles are present in the brains of age-matched non-demented individuals as in Alzheimer cases, but are absent in young controls lacking ANT. In extracellular "ghost tangles", the ultimate stage of neurofibrillary degeneration, immunoreactivity for tau is accessible to antibodies only when tissue sections are pretreated with formic acid to uncover the binding sites. In contrast to tau, presence/accessibility of an epitope residing on residues 50-65 of ubiquitin recognized by a monoclonal antibody raised to paired helical filaments (3-39) increases during the maturation of ANT and is most pronounced in "ghost tangles". Appearance/uncovering of the 3-39 epitope and masking of tau reactivity during tangle maturation may reflect degradation or conformational changes in the pathological filaments due to their aging and the final loss of their parent nerve cells. PMID- 2557643 TI - Ubiquitin and heat shock proteins in cultured nervous tissue after different stress conditions. AB - Ubiquitin (Ub) and Heat Shock Proteins (HSP) are believed to act jointly in the recovery process of an organism exposed to a stress. Ubiquitin has been recently found associated with paired helical filaments (PHF) in tangles of Alzheimer disease (AD) and Progressive Supranuclear Palsy (PSP) and with abnormal structures in other degenerative diseases, suggesting a role in their pathogenesis. To elucidate the mechanism of action, functions and relationship of Ub and the HSPs in the nervous tissue, we applied physical or chemical stress to neurons in culture. Rat dorsal root ganglia (DRG) explants were exposed either to aluminum lactate or to heat shock, the stress response was then studied with biochemical and immunological methods. The results show a clear dissociation in the formation of Ub-conjugates and the induction of the major HSPs, depending on the type of stress conditions. PMID- 2557645 TI - Mechanism of induction of long term potentiation by the mast cell degranulating peptide. AB - The properties of Long Term Potentiation (LTP), which is a sustained enhancement of the synaptic response produced by a brief train of electrical stimulation, and its mechanisms are briefly reviewed. Bath application of the mast cell degranulating peptide (MCD) - a peptide isolated from bee venom- also produces in hippocampal slices LTP which is indistinguishable from that produced by a train of electrical stimulation (no change in postsynaptic cell properties, APV sensitive). In vivo push-pull experiments indicate that this LTP is not associated with a sustained enhancement of release of glutamate and aspartate, the two principle transmitters of the hippocampus, but there is a delayed enhanced release of proteins in the extracellular space which may contribute to the maintenance of the potentiation. PMID- 2557646 TI - Effects of [D-Ala2] Met-enkephalinamide, a Met-enkephalin analog, on delayed response by squirrel monkeys. AB - Squirrel monkeys were tested on an indirect spatial delayed response task after subcutaneous injections of either physiological saline, 100, or 500 microgram/kg [D-Ala2] methionine enkephalinamide (DAME). During Experiment 1 monkeys completed two-choice delayed response problems with 0-, 6-, 12- and 18- sec delays; DAME significantly enhanced performance at 18-sec delays but not for the shorter delay. During Experiment 2, monkeys performed a nine-choice spatial delayed response task with 0-, 4- and 8-sec delays. In Experiment 2, low but not high doses of DAME impaired performance. Memory functions represented by a bias-free performance measure paralleled percentage correct in both experiments. Therefore, DAME effect, in enhancing and in impairing performance, were not attributable to changes in the relative number of systematic spatial errors (positional win shift; lose-stay, position perseveration, and position preference). PMID- 2557647 TI - The effect of portal and jugular infused glucose, mannitol and saline on food intake in dogs. AB - Hepatic glucoreceptors have been hypothesized to have an important role in determining normal hunger and satiety. In the present study 23 dogs were fitted with chronic hepatic portal and jugular vein cannulas. The dogs were fed for 1 hr/day. On infusion days (total of 318 infusions) the animals were infused into the portal or jugular veins with a 30% glucose solution (2.4 or 3.6 g/kg, b.wt.), 0.9% NaCl as a volume control or 30% mannitol as an osmotic control and then fed 10 minutes later. The data showed that the dog's food consumption was similar after they received glucose or the appropriate control infusion regardless of the infusion site. Some dogs had blood samples taken for glucose and insulin determinations prior to infusion, at the middle and end of infusion, just prior to food presentation and at the end of the feeding period. Saline and mannitol infusions did not alter plasma glucose or insulin concentrations; whereas there were marked increases in plasma glucose (6-8 x) and insulin (18-19 x) following glucose infusions. Postinfusion glucose values indicated approximately 72% of the infused dose glucose (approximately 43 g) had left the plasma prior to food presentation. Despite the large increases in plasma glucose and insulin, as well as glucose storage and/or oxidation, the dogs consumed amounts of food similar to that eaten after control infusions. Similarly, prefeeding the dogs 20% of their average daily intake prior to infusion did not alter the animals subsequent intake. These data are in agreement with earlier work from our laboratory and question the role of the hypothesized hepatic glucose satiety receptors. PMID- 2557648 TI - Affiliative behavior in voles: effects of morphine, naloxone, and cross fostering. AB - Species differences in affiliative behavior were examined in prairie and montane voles. Unfamiliar male-female pairs were placed in a test-cage for 2 hr and side by-side huddling was recorded during the third hour. Prairie vole pairs spent a mean of 31.2 minutes in contact whereas montane voles were in contact only 1.3 minutes. In order to examine the effects of experience on affiliative differences, pups of each species were cross-fostered. Fostered prairie vole parents did not survive longer than 7 days, whereas fostered montane voles were successfully weaned; cross-fostering had no effect on their huddling behavior when tested as adults. The effects of morphine (5 and 10 mg/kg) and naloxone (5 and 10 mg/kg) on side-by-side contact were evaluated in both species. Morphine (10 mg/kg) reduced huddling duration and activity levels in prairie voles. There were no other drug effects in either species. PMID- 2557649 TI - The topography of MK-801-induced locomotor patterns in rats. AB - Locomotor patterns in rats given systemic injections of the novel, noncompetitive N-methyl-D-aspartate (NMDA) antagonist, MK-801 were characterized using Digiscan Animal Activity Monitoring System. At low systemic doses, MK-801 produced an activity pattern most similar to patterns previously described for less potent noncompetitive NMDA antagonists; this was typified by hyperactive locomotor behavior, with increases in distance travelled, speed, and clockwise/anticlockwise locomotion, and a marked decrease in rearing behavior. Although MK-801 elicited some motor patterns similar to those previously described for sympathomimetic agents, including hyperactivity and increased stereotypy, it did not produce increased rearing behavior, the most prominent sympathomimetic effect. These results demonstrated that the topography of locomotion elicited by low systemic doses of MK-801 is most similar to locomotor patterns previously described for noncompetitive NMDA receptor antagonists. PMID- 2557650 TI - The ACTH (4-9) analog ORG 2766 in panic disorder: a preliminary study. PMID- 2557651 TI - Formation of PGI3 in the rat during dietary fish oil supplementation. AB - Conflicting results exist in the literature on the conversion of eicosapentaenoic acid (EPA) to trienoic prostaglandins and its influence on the formation of dienoic prostaglandins from arachidonic acid (AA). Tissues from animals fed fish oils produce little, if any, trienoic prostaglandins and reduced amounts of dienoic ones. Excretion of the major urinary metabolite of PGI2 is not reduced in humans taking fish oil, however, and substantial amounts of one derived from PGI3 have been found, by GC/MS. We have addressed this possible species difference by examining the urine of rats fed fish oil for 2.3 dinor-6-keto-PGF1 alpha and its delta 17 analog, formed from PGI2 and PGI3, respectively, and compared them with rats fed corn oil. Fatty acid differences in erythrocyte and aortic lipids were also determined. Rats fed fish oil do make PGI3 from eicosapentaenoic acid in vivo and do not suppress their production of PGI2, despite having more EPA than AA in aortic lipids. PMID- 2557652 TI - [Perspectives on chemotherapy of cancer of the bronchi]. AB - Chemotherapy of epidermoid bronchial cancer maintains, despite numerous therapeutic trails, a minimal efficacy and is shown to have no important effect on the duration of survival. Conversely, the immediate results obtained by chemotherapy for small cell carcinoma are remarkable particularly for the localised forms; there are numerous active drugs which can be usefully combined with radiotherapy to increase the level of complete remission and to attempt to influence the eventual outcome which remains unfavourable. However, this chemotherapy should benefit from the multiple research programmes which are currently underway to reinforce the effects of chemotherapy (new combinations of effective cytotoxic drugs, and the potential for changing biological responses) and to improve the techniques of application (for example, continuous infusions and chronomodulation). The intensification of chemotherapy (with or without the use of marrow transplants) seems to constitute, by the reduction of tumour mass that is induced, a factor determining the improvement in long-term results but is still limited only to small cell carcinomas. PMID- 2557653 TI - [Sulfidopeptide leukotrienes and asthma]. AB - The role of sulfidopeptide leukotrienes in asthma alone or in association with other mediators is still debated. Sulfidopeptide leukotrienes (s-LT) C4, D4 and E4 are 5-lipoxygenase derivatives of membrane arachidonic acid. All s-LT contract bronchial smooth muscle in vitro and provoke an acute bronchial obstruction in vivo in both healthy and asthmatic subjects. Numerous cells, including mast cells, alveolar macrophages, polynuclear basophils and eosinophils, are capable of secreting s-LT following such diverse stimuli as allergen exposure, platelet activating factor and calcium ionophore A 23187. In addition to constricting bronchial smooth muscle, s-LT promote the occurrence of mucosal oedema increase micro-vascular permeability in the bronchial wall, and cause hypersecretion of mucus by tracheo-bronchial glands. These effects lead to worsening of airways obstruction. s-LT increase non specific bronchial hyper-responsiveness. Clinical trials aiming to test the efficacy of new anti-leukotrienes are currently under way. In general these products have a real antagonistic effect on exogenous s-LT. Their efficacy vis-a-vis other types of stimulus such as allergens, histamine and exercise does not seem to be constant and depends largely on the composition and perhaps the route of administration (inhaled versus oral). The contribution of s LT as part of the therapeutic arsenal for the long term treatment of asthma remains to be established. PMID- 2557654 TI - [Small cell cancer of the bronchi: what is the role of surgery?]. AB - A series of 39 operated cases of small cell cancer were analysed. In 15 cases (38.4%) an exploratory thoracotomy only was carried out. 24 patients (61%) had a pulmonary excision: 12 pneumonectomies, 3 bilobectomies and 9 lobectomies were carried out. The hospital mortality was 7.7%. After a study of the surgical specimens these 21 cases were grouped as follows: 5 stage I, 1 stage II and 15 stage III. After surgery 4 patients were treated with radiotherapy alone, 3 chemotherapy alone and 10 by a combination of radiotherapy followed by chemotherapy, finally 4 patients had no post operative treatment. For the 21 patients who had resections the actuarial survival was 12.8% at 5 years and the mean survival 24 +/- 17.2 months. These results were judged as satisfactory and lead one to consider the current place of surgery in the treatment of small cell bronchial cancer. PMID- 2557655 TI - Hypothalamic pituitary adrenal function in patients with thalassemia major. AB - We examined the adrenocortical function in 14 italian thalassemic patients (8 f., 6 m.) aged 12 to 28. The following hormones were determined in each subject: plasma cortisol and aldosterone levels in both basal conditions and after maximal and submaximal stimulus (250 and 5 micrograms respectively) with synthetic corticotrophin beta 1-24 (Synacthen Ciba); corticotrophin and cortisol response to insulin-induced hypoglycaemia (0.15 U/kg iv.). Tests were repeated in all patients four years later. Normal controls were a group of 10 normal subjects matched for age, sex and body mass. Normal basal values of cortisol, aldosterone and ACTH were observed. Impaired cortisol response after stimulation with 5 micrograms of ACTH (6 patients) and after hypoglycaemia (4 patients) was identified. At the second test, four years later, one patient showed impaired cortisol response to both ACTH (5, micrograms) and hypoglycaemia, unlike the normal response to the first test. In all the others the cortisol response to stimulation did not differ from previous ones. In conclusion reduced ACTH and cortisol reserves detected in some thalassemic patients may be related to iron infiltration in the pituitary and adrenal glands. However, the present study did not indicate any significant correlation between either total blood load or serum ferritin and adrenal function parameters. Alteration in circulating hormones catabolism and impaired synthesis of transport proteins caused by chronic liver disease made adrenocortical hypofunction an even more complex picture to understand. PMID- 2557656 TI - The typical Reed-Sternberg phenotype and Ig gene rearrangement of Hodgkin's disease derived cell line ZO indicating a B-cell origin. PMID- 2557657 TI - Platelet-activating factor (PAF) stimulates phosphatidylinositol hydrolysis in human peripheral blood mononuclear leukocytes. AB - Human peripheral blood mononuclear leukocytes were labeled with myo-[3H]inositol and stimulated by platelet-activating factor (PAF). PAF-induced phosphatidylinositol hydrolysis and inositol-1,4,5-trisphosphate (IP3) formation in a dose-dependent manner. The response was very rapid and transient and parallels the time course of PAF-induced calcium mobilization in the same cells. However, the doses of PAF for inducing IP3 formation were much higher than those for calcium mobilization. PAF-induced IP3 formation was inhibited by PAF receptor antagonist, L-659, 989, but was not altered by either pertussis toxin or cholera toxin. These data suggest that PAF receptor in human peripheral blood mononuclear leukocytes may be coupled through a pertussis toxin-insensitive guanine nucleotide binding protein to a phosphoinositide-specific phospholipase C. PMID- 2557658 TI - Role of arachidonic acid metabolites in allergen-induced late responses. AB - The sheep model of allergic airway disease shares many pathophysiological similarities with allergic airway disease in humans. Studies performed in this animal model present strong evidence that the release of arachidonic acid metabolites plays an important role in the development of late bronchial responses to antigen challenge. The release of leukotrienes through the lipoxygenase pathway during the acute bronchial obstruction after inhalation of Ascaris suum antigen represents the key factor for the initiation of the subsequent events, namely the late phase response and the bronchial hyperreactivity. If this hypothesis can be substantiated in patients with bronchial asthma then pharmacologic modification of the lipoxygenase pathway and/or products may be important in the treatment of asthma. PMID- 2557659 TI - Ventilatory responses to hypoxia in healthy subjects: a comparison between young children and adults. AB - Ventilatory responses and breathing patterns during acute hypoxia have been studied at sea level in healthy subjects (8 children, 8 adults). While breathing ambient air it appears that, compared to adults, children hyperventilate when ventilation is standardized to body mass unit. During hypoxia, ventilatory strategy differs with age: when adults essentially increase tidal volume (VT), children increase VT and ventilatory frequency (fr). The ventilatory steady-state response to hypoxia is lesser in children than in adults, that is to say, children have a lower O2 ventilatory sensitivity. These results, which show that adults and children have qualitatively and quantitatively different ventilatory responses to hypoxia, are interpreted in terms of the ability to displace gaseous volumes. PMID- 2557660 TI - Effect of captopril and enalapril medication on the serum ACE test for sarcoidosis. AB - The presence of an intrinsic inhibitor of ACE in blood of approximately 25% of sera submitted for serum ACE assay in the diagnosis and evaluation of patients with sarcoidosis, and the common use of ACE inhibitors (captopril and enalapril) in the treatment of hypertension and congestive heart failure, stimulated these studies to compare the effects of the intrinsic and medicinal inhibitors upon serum ACE activity. Since the intrinsic ACE inhibitor in man is affected by serum dilution (inhibition is reversed) and by dialysis (inhibition becomes irreversible), these manipulations were also studied with medicinal inhibitors to provide guidelines for suspecting their presence in submitted serum samples without an accompanying history. Enalapril was found to have delayed onset of action after oral administration (1-2 hours), and was even further delayed (4 hours) when the intrinsic inhibitor also happened to be present. Inhibition by enalapril was not attenuated with refrigerated storage, with dilution or with dialysis of the serum. Captopril had a more rapid time of onset of ACE inhibition, but its inhibitory activity was markedly reduced with refrigerated storage of the serum; patients showed either a short half-life for the effect (1 4 days) or a prolonged half-life (10-17 days). Inhibitory activity of captopril was reversed following dilution of serum or following dialysis. The reversal of inhibition by captopril following dialysis, therefore, differed from the effect of dialysis on the intrinsic ACE inhibitor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557661 TI - Pulmonary masses in a 37 year-old man. PMID- 2557662 TI - Overuse syndromes in instrumental musicians. PMID- 2557663 TI - Polymorphonuclear neutrophils: an effective antimicrobial force. AB - The production and deployment of polymorphonuclear neutrophils (PMNs) are under close regulation. PMNs interact through cytokines with a number of cell types, including macrophages, lymphocytes, and endothelial cells. PMNs are guided by bacterial products and cytokines to target sites, where microbes are recognized and killed. Killing occurs through oxygen-dependent and oxygen-independent mechanisms. The frequent and severe infections seen in patients with defects (either congenital or acquired) in PMN function demonstrate the importance of PMNs in host defense against infection. PMNs are potent inflammatory cells and can exacerbate disease states such as myocardial ischemia, gram-negative bacterial sepsis, and the adult respiratory distress syndrome. PMID- 2557665 TI - [Adenomas of the liver and oral contraception. Clinical and therapeutic aspects apropos of a giant adenoma]. AB - The first cases of benign liver tumors following estroprogestative therapy were described in 1973. Since that time, this relation between cause and effect has been demonstrated. The authors report the case of a giant liver adenoma which developed after 13 years of uninterrupted use of oral contraceptives. The advantage of this disease is not so much its prevalence which is low, but its clinical diagnosis. In fact, failure to recognize this entity may result in dramatic complications such as haemorrhagic rupture of the liver. The therapeutic approaches, whether wait-and-see or aggressive, all present a common objective: permanent discontinuance of estroprogestative drugs. PMID- 2557664 TI - Prevention of cytomegalovirus infection after marrow transplantation. AB - Primary infection with cytomegalovirus (CMV) in seronegative patients is acquired most commonly from blood products or marrow taken from seropositive donors, whereas recurrent infection in seropositive patients appears to be derived predominantly from reactivation of latent endogenous virus. Not all infected patients develop symptomatic CMV disease; the factors that determine severity of infection and recovery are not yet fully defined. The epidemiology of CMV infection indicates that primary infection is preventable by use of seronegative blood products, including marrow. Such techniques should be implemented immediately. Leukocyte depletion of blood products is a potential alternative, but both efficacy and logistics of implementation require further study in patients who need large quantities of blood products. The efficacy of passive immunoprophylaxis with immunoglobulins remains uncertain, and this modality cannot be recommended until additional data are available. Among seropositive patients, antiviral agents may be used to suppress CMV infection until periods of improved immunocompetence. Although not effective for the treatment of established CMV infection, intravenous acyclovir significantly reduced the probability of CMV infection and disease and improved survival in a controlled trial. The newer antiviral agents ganciclovir and foscarnet should provide better protection, although the marrow toxicity of ganciclovir is problematic. Future directions will include attempts to restore specific immune responses with subunit or recombinant vaccines and adoptive immunotherapy with CMV-specific effector cells generated in vitro. PMID- 2557666 TI - [Uterine perforation revealing a rare placental tumor: trophoblastic tumor of the implantation site. Review of the literature. Apropos of a case]. AB - The trophoblastic tumor of the implantation site (TTIS), recently discovered, is a peculiar and rare choriocarcinoma; the authors are reporting the 36th case in the world. Clinically, it is different from the typical choriocarcinoma by amenorrhea, frequent uterine perforations and negative pregnancy tests. The diagnosis is made histologically; most important are the monomorphic tumor cells, and especially, the immunocytochemical labelling for human chorionic gonadotropic hormone and lactogenic placental hormone permitting the differentiation of TTIS in choriocarcinoma from TTIS in non-trophoblastic tumors. Its course is highly malignant in 25 p. cent of the cases without any absolute prognostic factor at this time. The treatment is not codified; it seems that, unlike in the typical choriocarcinoma, a place of choice should be given to surgery, since the usual chemotherapy of trophoblastic tumors has not proved to be effective in this indication. On the contrary, monitoring will be supported by serum hCG assays. PMID- 2557667 TI - Screening for hereditary colorectal carcinoma. PMID- 2557668 TI - Omeprazole: mode of action and effect on acid secretion in animals. AB - H+,K+-ATPase constitutes the final step in the acid secretory process that takes place in the parietal cell, and this enzyme has recently been recognized as a target for inhibitors of acid secretion. The gastric H+,K+-ATPase is located in the uniquely acidic environment of the parietal cell. Omeprazole, a weak base, is concentrated within the acid canaliculus of the parietal cell and is rapidly converted to an inhibitor of the H+,K+-ATPase in the acid compartments of the parietal cell. Omeprazole is thus activated close to its target enzyme, where it is present in high concentrations, and binds selectively to the gastric H+,K+ ATPase. Omeprazole is the first H+,K+-ATPase inhibitor to be used in the treatment of acid-related diseases, and inhibition of acid secretion is closely correlated to inhibition of gastric H+,K+-ATPase activity. Both basal and stimulated acid secretion are inhibited by omeprazole in a dose-dependent manner, irrespective of the nature of the stimulus. Omeprazole exerts a prolonged antisecretory effect, and in dogs, 4 days are required for return to normal acid secretion following a maximal inhibitory dose. PMID- 2557669 TI - The gastric H+,K+-ATPase: the site of action of omeprazole. AB - The mammalian parietal cell is dedicated to the secretion of HCl in response to various stimuli and second messengers. Oxidative metabolism in the cell increases about 10-fold in order to supply ATP to the gastric proton pump, the H+,K+ ATPase. This pump appears to be present only in the parietal cell. This membrane embedded enzyme uses the scalar energy of ATP hydrolysis to carry out the vectorial transport of H+ in one direction in exchange for K+ in the other direction. In the cytoplasmic vesicle, K+ does not permeate the membrane, whereas in the secretory canaliculus, there is a Cl- channel and a KCl cotransport pathway which allow K+ and Cl- to exit from the cell. The K+ is then recycled back into the cell by the ATPase, and H+ secretion occurs into the canalicular space. Although there are other proton pumps, only the gastric H+,K+-ATPase has this exchange mechanism and only the gastric H+,K+-ATPase is able to generate a pH of less than 4. Thus the gastric proton pump has a unique structure and mechanism, and produces a unique luminal pH. This enzyme is therefore an appropriate target for rational drug design. PMID- 2557670 TI - Omeprazole in the treatment of duodenal ulcer. AB - Omeprazole is the most effective antisecretory agent available today. Open and dose-comparative studies have documented that at dosages of 20 mg/day or more, the drug produces duodenal ulcer healing rates of 90-100% after 4 weeks. Controlled trials show that omeprazole, 20-40 mg/day, is superior to cimetidine and ranitidine in healing duodenal ulcer, with a median therapeutic gain of 21% at 2 weeks and 15% at 4 weeks. Ulcer symptom relief is also more pronounced and faster with omeprazole than with H2-receptor antagonists. No significant side effects attributable to treatment with omeprazole have appeared in any of these studies or in the accumulated experience from several thousand patients treated with omeprazole. No tendency to an increase in recurrence rate after discontinuation of treatment with omeprazole has been shown. In summary, omeprazole constitutes a major advance in the short-term treatment of duodenal ulcer, giving fast and pronounced healing and symptom relief. PMID- 2557671 TI - Nonclassical organ involvement in temporal arteritis. PMID- 2557672 TI - [Effects of intracisternal injection of naloxone on the depressor response to stimulation of renal afferent nerve in rabbits]. AB - The purpose of this study was to observe the effects of opioid receptor and alpha adrenergic receptors in the lower brain stem on the depressor response to electrical stimulation of renal afferent nerve, using intracisternal injection of blockers of these receptors. Experiments were conducted in sodium pentobarbital anesthetized rabbits. The intracisternal injection (ict) of artificial cerebrospinal fluid (CSF) did not greatly affect the depressor response to stimulation of renal afferent nerve (RAS) and of aortic nerve (ANS). Ict of 550 nmol naloxone significantly inhibited the depressor response to RAS (P less than 0.05) but enhanced the depressor response to ANS (P less than 0.05). Ict of 335 nmol phentolamine significantly inhibited the depressor response to both RAS and ANS. Ict of naloxone reversed the phentolamine inhibition of the depressor response to ANS but did not affect the phentolamine inhibition of the depressor response to RAS. Ict of both naloxone and phentolamine did not affect the inhibitory interaction between the RAS-induced depressor and aortic baroreflex. Ict of phentolamine significantly decreased MAP, and ict of naloxone did not decrease MAP but reversed the decrease in MAP by ict of phentolamine. These data suggest that the activation of opioid receptors in the lower brain stem enhances the depressor response to RAS and attenuates the depressor response to ANS. Moreover, activation of alpha-adrenergic receptors in the lower brain stem facilitates the depressor response to RAS and ANS. PMID- 2557673 TI - [Effects of kappa-opiate receptor antagonist MR-2266-BS on ACTH and prolactin release]. AB - The effect of intravenous injection of different doses of MR-2266-BS, a selective antagonist of kappa-opiate receptor, on plasma adrenocorticotropin (ACTH) and prolactin (PRL) in conscious male rats bearing an intrajugular cannulae was assessed. The results revealed that the MR-2266-BS of 3 mg/kg completely blocked the restraint stress-induced increase in plasma ACTH levels, and further elevated plasma PRL levels in these animals, while there were no effects on the resting levels of ACTH and PRL. MR-2266-BS of 6 mg/kg significantly increased the resting levels of plasma ACTH and also further elevated the restraint stress-induced increase of plasma ACTH and PRL. The present data suggest that kappa-opiate receptor and its endogenous ligand may be involved in the regulation of the resting and restraint stress-induced release of ACTH, and their action appears to be both stimulatory and inhibitory. Furthermore, kappa-opiate receptor and its endogenous ligand may only inhibit the stress-induced release of PRL. PMID- 2557674 TI - [Effects of aliphatic alcohols on slow action potential of guinea-pig papillary muscle]. AB - Effects of ethanol, butanol, hexanol and octanol on slow action potential (APA, APD50, Vmax, and V'max) and its correspondent ICa (Imax) and Ik (I'max) of guinea pig papillary muscle were measured and analysed by using phase-plane method. The results showed that all of four alcohols inhibited calcium current of cardiac membranes, but the action on potassium current changed from enhancement (ethanol) to impairment (hexanol and octanol) with the increase in carbon number of alcohols. Furthermore, the longer the chains of alcohols, the less the concentration needed to produce these effects. This phenomenon suggests a possible correlation between the actions of aliphatic alcohols and their hydrophobic properties. PMID- 2557675 TI - [Changes of Isi and Ix in sheep cardiac Purkinje fibers during alpha- and beta- adrenoceptor excitation]. AB - When alpha- and beta- adrenoceptors were activated respectively in sheep cardiac Purkinje fibers, changes of the slow inward current (Isi) and delayed rectifier current (Ix) were studied by using two-microelectrode voltage clamp technique. When beta-receptor was blocked by propranolol, phenylephrine 5.0 x 10(-6) mol/L increased the peak value of Isi from 17.5 to 26 nA (n = 6, P less than 0.05) and prolonged the ms recovery time of Isi from inactivation from 293 +/- 51 ms to 441 +/- 190 (n = 4, P less than 0.05). Phenylephrine had no effect on the delayed rectifier current (Ix). When alpha-adrenoceptor was blocked by phentolamine, isoprenaline of 4 x 10(-7) mol/L increased the peak value of Isi from 27.7 to 40.8 nA (n = 7, P less than 0.05) and had no effect on the kinetics of Isi, but it did increase the tail current (Ix) peak value from 6.7 to 14.4 nA (n = 6, P less than 0.05). The results suggest that alpha- and beta-adrenoceptor agonists differ in their effects on delay rectifier current. PMID- 2557676 TI - [Effect of myocardial alpha 1-adrenoceptor stimulation on the guinea-pig ventricular papillary muscle]. AB - The alpha-adrenoceptor agonist phenylephrine (5.0 x 10(-6) mol/L) was used to stimulate myocardial alpha-adrenoceptors of the guinea-pig ventricular papillary muscle, and changes of transmembrane action potential and contractile force of the muscle were observed. The alpha 1-adrenoceptor blocker prazosin (5.0 x 10(-7) mol/L) and the alpha 2-adrenoceptor blocker yohimbine (5.0 x 10(-7) mol/L) were used to determine which subtype of alpha-adrenoceptor is responsible for the effects. The beta-adrenoceptor blocker propranolol (1.0 x 10(-6) mol/L) was used throughout the experiment. The results show that the myocardial alpha 1 adrenoceptor stimulation (1) increases the contractile force of the guinea-pig ventricular papillary muscle, (2) prolongs the time to peak contractile force while the duration of relaxation is not altered, (3) prolongs the fast response action potential duration, and (4) increases the maximal rate of depolarization during the phase 0 of the slow response action potential. It is suggested that the electrophysiological and positive inotropic effects of myocardial alpha 1 adrenoceptor stimulation might be due to the activation of the slow inward current and an increase in Ca2+ influx. PMID- 2557678 TI - Significance of intraoperative ultrasonography in anterior spinal operation. AB - Intraoperative spinal ultrasonography (IOSU) was performed during anterior spinal operations in 124 patients. The IOSU proved very useful in ascertaining whether the bone-removing width was sufficient for the spinal transverse diameter. It was also possible to check intraoperatively for unremoved material that was still compressing the cord (in particular, posterior lateral osteophytes and hernial prolapse). When the insufficient width of bone removal or remaining compression factor was found, an additional technique could be performed immediately, and the IOSU was helpful for prevention of cervical reoperation and improvement of surgical results. The degree of spinal pulsations observed in IOSU was of great value in the prediction of the prognosis for spinal cord postoperatively. PMID- 2557677 TI - Postoperative instability of cervical OPLL and cervical radiculomyelopathy. AB - The presence of cervical spine instability with respect to preoperative and postoperative changes in angular, horizontal, and rotational displacement of the vertebral body were studied. With the anterior approach, the instability in the remaining unfused segments, and their relation to the kyphotic or lordotic fused segment were studied. With the posterior approach, postoperative ROM (range of motion) could be better maintained, and horizontal displacement was improved in more cases by laminoplasty compared with laminectomy. With the anterior approach, the compensatory function for the loss of motion of the segments resulting from fusion was most remarkable at the levels of C2-3 and C6-7. In the alignment of the anterior fused segments, it appears important that the physiologic lordotic position be maintained. PMID- 2557679 TI - Exploring the effect of a spinal manipulation on plasma beta-endorphin levels in normal men. PMID- 2557680 TI - [Polishing of titanium prosthetics (Part 6). The chemical polishing baths containing hydrofluoric acid and nitric acid]. AB - Titanium was polished using several chemical polishing baths containing different ratios of hydrofluoric acid and nitric acid. The meltage, surface roughness, and surface texture of titanium samples after chemical polishing were affected by the ratio of hydrofluoric acid and nitric acid. Generally the meltage increased and surface roughness decreased when the mole concentration of hydrofluoric acid was high and that of nitric acid was low. For example the chemical polishing bath containing 5 mole hydrofluoric acid and 5 mole nitric acid improved the surface texture in one minute, but SEM observation revealed a partially rough surface caused by the excessive solution. The chemical polishing bath containing 1 mole hydrofluoric acid and 5 mole nitric acid did not improve the surface texture in a short time because of low solubility, but improved the surface texture gradually with the extension of the immersion time and a good surface texture was observed by SEM. The chemical polishing using the chemical polishing bath with low solubility and immersion of the prosthetics for a rather long time were considered useful procedures to obtain a smooth surface of titanium prosthetics while maintaining their accuracy. PMID- 2557681 TI - [Self-setting apatite cement. VII. Barium-apatite as radio-opaque medium]. AB - Addition of barium hydroxyapatite (BaAp) successfully bestowed clinically acceptable radioopacity to the self-setting apatite cement consisting of an equimolar mixture of tetracalcium phosphate and dicalcium phosphate dihydrate. To accelerate the setting reaction which was retarded by Ba2+ released from BaAp at a lower pH during first stage of spatulation with 20 mM phosphoric acid, calcium hydroxyapatite (CaAp) was added to the cement mixture. At about 20 wt% of BaAp and 20 wt% CaAp, the setting reaction proceeded at a neutral or weak alkaline pH, which is one of the most promising aspects of the self-setting cement and assures that this type of cement may be the least irritating of the dental cements presently available. The cement spatulated at L/P = 0.4 set within 10 minutes and its radiopacity was comparable to or more than that of tooth enamel. The wet compressive strength of the set cement stored for one day in synthetic saliva at 37 degrees C was approximately 100 kgf/cm2. Although this value is almost one fourth that of 40 wt% CaAp cement, this cement appears to be strong enough to apply as root canal filling material. PMID- 2557682 TI - [Computer graphics of apatite crystals]. AB - Hydroxyapatite was studied by computer graphics. The data of structural coordinates of hydroxyapatite crystal was put into a protein graphics program in A units. The connection of each element, with what comes front and rear, was displayed by the shade-line erasing method, and the solid-image was expressed by the degree of lightness from the light source. The tint of color was drawn by Raster graphics method. A fine picture can be obtained with the program. Rotating the graphics freely in an angle around X, Y or Z axis, gives a view from any direction. Crystal graphics model of fluoridated hydroxyapatite, in which F- ions substituted in part into OH- position, was also drawn. PMID- 2557683 TI - [Effect of filler system on the mechanical properties of light-cured composite resins. I. Effect of various types of silica fillers on the mechanical properties of the composite resins]. AB - To investigate the effect of silica fillers on the mechanical properties of visible light-cured composite resins, Bis-GMA-based composites with four types of silica fillers were prepared. The mechanical properties of the composites with splinter-shaped silica fillers increased with increasing the filler fraction. Although the spherical silica filler could be filled more with resin monomer than the splinter-shaped silica filler, the mechanical properties of spherical silica filled composite were relatively lower than those of the composite with splinter shaped silica fillers. The micro particle silica-filled composite showed no obvious increase in the mechanical properties in either the dry or wet conditions. SEM observations of the fractured resin surface revealed that fracture occurred through the resin matrix as well as the resin/filler interface and the mechanical properties of each composite resin were correlated with the nature of crack propagation. PMID- 2557684 TI - [Effect of filler system on the mechanical properties of light-cured composite resins. II. Mechanical properties of visible light-cured composite resins with binary filler system]. AB - To improve the fracture resistance of the composite resin system under highly stressed conditions, a variable amount of microfiller was incorporated into the light-cured composite resins containing splinter-shaped silica filler. With the increasing the microfiller content (10-30 wt%) in the resin matrix, the elastic modulus, compressive proportional limit and compressive fracture strength of the composite resin were increased. The plastic deformation of the composite resins under compressive stress were decreased with increasing microfiller content. These findings suggested that the combination of a larger silica filler and microfiller would increase the compressive fracture resistance of the composite resin systems. PMID- 2557685 TI - [An in vitro study of dentin hypersensitivity using calcium phosphate cement]. AB - Calcium phosphate remineralizing slurry (CPRS), consisting of an equimolar mixture of tetracalcium phosphate (TTCP) and dicalcium phosphate dihydrate (DCPD), has previously been shown to form hydroxyapatite (HAp) spontaneously at physiological pH. Since considerable amounts of HAp may be formed in situ by CPRS, it may be effective for desensitizing hypersensitive dentin by obturating exposed dentinal tubule openings. In the present study, the effects of topically applied CPRS to etched dentin surfaces were investigated. The samples were randomly divided into 3 groups: (1) the control specimens were soaked in a saliva like (SL) solution (2) the specimens received either 10-minute or 1-hour treatment with CPRS and (3) the specimens were treated with viscous CPRS, containing carboxymethyl cellulose (CMC) gel, for 8 hours. The SEM observations revealed that the control specimens were covered with a thin layer of plate-like crystals indicative of octacalcium phosphate (OCP). No significant penetration of the crystals into the dentinal tubules was detected. In CPRS group, the 10-minute treatment produced a 10 to 15 microns thick dense layer of precipitation consisting of needle-, rod-, and plate-like crystals. The 1-hour treatment produced a similar precipitation except that the crystals were mostly needle like. A cross section view of the samples indicated remarkable penetration of the crystals into the tubules, thus providing significant obturation of tubule openings. Samples in the CMC-gel group were covered with an even more impervious layer of precipitation consisting of crystals of a variety of morphologies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557687 TI - Post-mortem stability of thyrotropin-releasing hormone and muscarinic cholinergic receptors in rat forebrain. PMID- 2557686 TI - Mapping cocaine binding sites in human and baboon brain in vivo. AB - The first direct measurements of cocaine binding in the brain of normal human volunteers and baboons have been made by using positron emission tomography (PET) and tracer doses of [N-11C-methyl]-(-)-cocaine ([11C]cocaine). Cocaine's binding and release from brain are rapid with the highest regional uptake of carbon-11 occurring in the corpus striatum at 4-10 minutes after intravenous injection of labeled cocaine. This was followed by a clearance to half the peak value at about 25 minutes with the overall time course paralleling the previously documented time course of the euphoria experienced after intravenous cocaine administration. Blockade of the dopamine reuptake sites with nomifensine reduced the striatal but not the cerebellar uptake of [11C]cocaine in baboons indicating that cocaine binding is associated with the dopamine reuptake site in the corpus striatum. A comparison of labeled metabolites of cocaine in human and baboon plasma showed that while cocaine is rapidly metabolized in both species, the profile of labeled metabolites is different, with baboon plasma containing significant amounts of labeled carbon dioxide, and human plasma containing no significant labeled carbon dioxide. These studies demonstrate the feasibility of using [11C]cocaine and PET to map binding sites for cocaine in human brain, to monitor its kinetics, and to characterize its binding mechanism by using appropriate pharmacological challenges. PMID- 2557689 TI - PCR and the cloning of receptor subtype genes. PMID- 2557688 TI - Selective inhibition of polymorphonuclear neutrophil activity by 2,3,7,8 tetrachlorodibenzo-p-dioxin. AB - Although the environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), via its interaction with the Ah receptor, is an extremely potent carcinogen and immunosuppressive agent in experimental animals, its possible actions on polymorphonuclear (PMN) function have not been determined. In addition to their importance against infectious organisms, PMNs have been implicated in antitumor resistance. The present studies examined the effects of in vivo exposure to TCDD on PMN function in B6C3F1 (TCDD sensitive, presence of high affinity Ah receptor) and DBA/2N (TCDD resistant at low doses, defective Ah receptor) mice. Animals received a single oral exposure of 5 or 10 micrograms/kg of TCDD and PMNs were obtained 5 days later from the peritoneal cavity following elicitation with sodium caseinate. TCDD reduced the cytolytic and cytostatic activity of PMA-activated PMNs in B6C3F1, but not in DBA/2N mice, suggesting that this response segregates with the Ah locus. Furthermore, TCDD was found to bind specifically to PMNs from Ah-responsive mice. Neither the production of superoxide and hydrogen peroxide nor degranulation, the latter measured by beta glucuronidase release, was impaired. Supernatants recovered from PMN cell cultures of TCDD-sensitive mice, but not from resistant DBA/2N mice, showed reduced killing capacity for actinomycin D-treated L929 tumor cells, while their ability to bind to tumor cells was not altered. These data suggest that TCDD interferes with PMN-mediated tumor cell killing by altering the production or secretion of a cytolytic factor. Examination of bone marrow stem cells revealed that granulocytic but not monocytic colonies were reduced after TCDD exposure in vivo and in vitro. Although mature PMNs had detectable levels of Ah receptor, exposure in vitro of these cells to TCDD had no effect on antitumor activity. Thus, it is possible that TCDD may affect PMNs at the level of hematopoiesis, via a direct interaction with granulocyte precursor cells, or modulate PMNs at different stages of maturation. PMID- 2557690 TI - Hormone and growth factor receptor-mediated regulation of phospholipase C activity. AB - The broad importance of receptor-activated phosphoinositide hydrolysis in the physiological action of hormones, neurotransmitters and growth factors has sparked interest in the study of transmembrane signalling events responsible for activation of phospholipase C. As with receptors involved in regulation of adenylyl cyclase, ion channels and phototransmission, it is clear that a guanine nucleotide regulatory protein (G protein) is a necessary component of the hormone and neurotransmitter-regulated phosphoinositide signalling mechanism. Recent evidence to support a possible second mode of regulation of phospholipase C by growth factor receptors is emerging in the form of realization that at least one isozyme of phospholipase C serves as a substrate for the tyrosine kinase activity of growth factor receptors known to stimulate phosphoinositide hydrolysis. In this review, Jose Boyer and colleagues summarize progress towards delineating the properties and identity of the G protein(s) involved in this pathway, recent advances in purification and molecular cloning of phospholipase C isozymes, and the current understanding of growth factor receptor-mediated regulation of phosphoinositide hydrolysis. PMID- 2557691 TI - Bilateral bronchioloalveolar carcinoma of the lungs in a 7 year old girl treated for Hodgkin's disease. AB - In a 7 year old girl who had previously been given intensive treatment for Hodgkin's disease with numerous courses of multiagent therapy including MVPP (nitrogen mustard, vinblastine, prednisone, procarbazine) and who received radiotherapy in a total dose of 32 Gy to the sternal, and 24 Gy to the cervical (left and right) and axillary (left and right) regions, an autopsy revealed bilateral bronchioloalveolar carcinoma of the lungs. PMID- 2557692 TI - Cytogenetic analysis in 28 radically operated non-small-cell lung cancer: preliminary considerations. AB - Twenty-eight radically operated non-small-cell lung cancer patients were analyzed with regard to chromosomal assessment and DNA content: in 13 cases, different quantitative/qualitative chromosome alterations were found. In particular, in 12 cases marker chromosomes and cytogenetic abnormalities in euploid cells were demonstrated. The prognostic value of these findings will be aim of further studies. PMID- 2557693 TI - Phase II trial with alternating two drug schedules, CAP/MEC', for advanced (stage III Mo/M1) non-small-cell lung cancer. AB - Sixty-eight evaluable patients with advanced squamous cell carcinoma (48), large cell carcinoma (2) and adenocarcinoma (18) of the lung were treated with a six drug regimen delivering two monthly alternated combinations. The combinations were cisplatin, adriamycin and cyclophosphamide (CAP) and methotrexate, etoposide and CCNU (MEC'). Following a minimum of two courses, the overall response rate was 22% (confidence limits, 12% to 32%) (15/68, 2 complete responses and 13 partial responses); 47% (32/68) had stable disease and 31% (21/68) had progressive disease. The responses lasted a median of 3 months (range, 1-15 months). The actuarial median survival was 11 months in responsive patients, 10 months in stable disease patients, and 5 months in progressive patients. The overall median survival obtained was 9 months (range, 2-28+ months). Toxicity was minimal, and subjective tolerance of the treatment appeared good. However, this alternating program did not improve response rate or survival. PMID- 2557695 TI - Evidence for altered hypothalamic-hypophyseal-gonadal axis in untreated patients with testicular germ-cell tumor. AB - To investigate the function of the hypothalamic-hypophyseal-gonadal axis in testicular germ cell tumors, we evaluated gonadotropin responses to gonadotropin releasing hormone (GnRH) in 12 untreated patients with testicular cancer (5 seminomas and 7 non-seminomas). GnRH was given i.v. at a dose of 100 micrograms as a bolus, and venous blood samples were collected at 0, 20, 60, and 120 min. As controls, 14 healthy males were studied. Basal levels of testosterone, estradiol and prolactin were also detected in each patient. Hormonal serum concentrations were measured by the radioimmunoassay. Mean basal testosterone, estradiol and prolactin levels were not significantly different from those of controls. Patients had a lower FSH and LH peak after GnRH than controls, without, however, any significant difference. As regards histology, non-seminoma patients lacked an FSH response to GnRH and had statistically lower mean peak levels than controls. Moreover, non-seminoma patients had statistically lower mean peak values of LH after GnRH than controls. These data show that patients with testicular germ cell tumor, and more particularly those with non-seminomas, have an altered function of the hypothalamic-hypophyseal-gonadal axis, which is already present prior to therapy. Further studies, particularly in stage I patients treated only with orchiectomy, should be performed to confirm and better define the physiopathologic significance of the altered hypothalamic-hypophyseal-gonadal axis in testicular cancer and to clarify the alteration of fertility, which is frequently present before treatment. PMID- 2557694 TI - Serum copper and ceruloplasmin in early and in advanced hepatocellular carcinoma: diagnostic and prognostic relevance. AB - To evaluate the prognostic value of serum copper (S-Cu) and ceruloplasmin and their pathophysiologic significance in human hepatocellular carcinoma (HCC), we studied 49 patients with HCC (20 of which were submitted to partial hepatectomy) compared with 110 patients with liver cirrhosis. In HCC both S-Cu and ceruloplasmin were higher than in cirrhosis; moreover, S-Cu was correlated with the extension of HCC, evaluated by instrumental data and by surgical inspection. In cirrhotic patients, mean S-Cu was 122.9 micrograms/dl (SD, 29.3), in early HCC, 153.0 micrograms/dl (SD, 34.5), and in advanced HCC, 193.1 micrograms/dl (SD, 37.7). Variance analysis gave F = 59.4. In HCC patients S-Cu was positively correlated with ceruloplasmin and with fibrinogen. Survival, evaluated by Mantel's test stratified for surgical therapy, was longer in patients with S-Cu levels lower than 175 micrograms/dl and in those at an earlier stage. We therefore conclude that S-Cu has a relevant diagnostic value in detecting HCC also in early stage and allows prognostic evaluation as regards survival. PMID- 2557697 TI - Effects of hydrostatic pressure and inert gases on twitch tension. AB - Effect of pressure and inert gases on the twitch tension (Tmax) was measured on electrically stimulated and spontaneously beating rat atria. In stimulated preparations, pressurization to 10 MPa increased Tmax by 20-60% depending on the stimulating frequency (60-240 beats/min). The introduction of 5 MPa N2 or 5 MPa H2 at 10 MPa hydrostatic pressure decreased the Tmax by 17 +/- 6% and 13 +/- 6%, respectively. Gas effect did not depend on the stimulating frequency. Nitrous oxide (0.15 and 0.45 MPa) decreased Tmax both at "surface" and at 10 MPa. Nitrous oxide effect was slightly potentiated at pressure. In spontaneously beating preparations, compression to 10 and 15 MPa decreased beating frequency (BF) by 24 +/- 10% and 31 +/- 8% and increased Tmax by 60 +/- 35% and 105 +/- 33%, respectively. The tension increase is partly due to the direct pressure effect and partly due to the negative force-frequency relation in the rat atria. Introduction of inert gas increased BF and decreased Tmax. The potency of the gases was in the same order for both variables: He less than H2 less than N2. PMID- 2557696 TI - In situ and microinvasive carcinoma with squamoid differentiation arising in a phyllodes tumor: report of a case. AB - A case of in situ and invasive ductal carcinoma with areas of squamoid differentiation arising in a phyllodes tumor is reported. This last aspect has never been described before and is discussed reviewing the previously reported cases in the literature. PMID- 2557698 TI - [Feasibility and relevance of tumor volumetry for stage classification and assessment of remission of germ cell tumors]. AB - Although tumor load has proven to be the most relevant prognostic factor in disseminated germ cell tumors (GCT), methods to determine tumor volume for staging have not been studied so far. In a prospective study, we therefore measured the volume of metastases before and during chemotherapy in 27 patients with disseminated GCT. Abdominal tumor volume was calculated using a General Electric CT scan 8800. Total volume was determined by cumulation of 1 cm slices measured by a cursor. Pulmonary volume was calculated by taking each metastasis as a sphere using V = 0.523 x d3, where V = volume and d = diameter. We used linear regression analysis to determine the dependence of tumor markers on volume. Before chemotherapy, the median tumor volume of all patients was 237 (range 4-2690) cm3. The tumor volume was 1-100 cm3 in 30%, 101-500 cm3 in 41%, and over 500 cm3 in 29% of the patients. NED (no evidence of disease) was achieved in 8/8 patients presenting with a small (1-100 cm3) and 9/10 with a moderate (101-500 cm3) tumor volume. In contrast, only 1/8 with advanced tumor load (greater than 500 cm3) achieved NED. While there was a significant correlation between the initial and the residual tumor volume (P = 0.0024, r = 0.72), there was none between the tumor volume and alpha fetoprotein, beta human chorionic gonadotropin, and lactate dehydrogenase. These results suggest that radiological determination of tumor volume is a reproducible and accurate staging method. PMID- 2557699 TI - [Resection of the right lobe of the liver in children with liver tumors]. PMID- 2557700 TI - Adenoviral hepatitis in a merlin (Falco columbarius). PMID- 2557701 TI - Winter dysentery in adult dairy cattle. PMID- 2557702 TI - Detection and survival of group A rotavirus in a piggery. AB - Samples of dust, faeces and effluent were collected from a piggery and examined for group A rotavirus, using a commercial ELISA test, electron microscopy and inoculation of MA-104 cells. Rotavirus antigen was demonstrated in samples collected from farrowing and weaner rooms but not from fattener and sow houses. Rotavirus antigen was also detected in samples collected from a weaner room which had been free of piglets for three months. A cytopathic porcine rotavirus (British isolate SW20/21) was kept at room temperature for four months; it survived with titres reduced by 2 log10. These observations suggest that the environment of commercial piggeries is an important source of rotaviral infection for young piglets. PMID- 2557703 TI - Bovine encephalitis herpesvirus. PMID- 2557704 TI - Persistent polyuria in two dogs following adrenocorticolysis for pituitary dependent hyperadrenocorticism. AB - In two dogs with pituitary-dependent hyperadrenocorticism, adrenocorticolysis with o.p'-DDD led to the disappearance of the signs and symptoms except for the polyuria. After a modified water-deprivation test the osmoregulation of vasopressin release was studied by hypertonic saline infusion. In both dogs the hypertonicity, thus induced, resulted in very minimal responses of the vasopressin secretion. PMID- 2557705 TI - Effects of polymyxin B on selected features of equine carbohydrate overload. AB - Gram negative endotoxins play a contributory role in the syndrome which results from over consumption of carbohydrates by horses and ponies. Since the antibiotic polymyxin B exerts a direct anti-endotoxin effect by chemically modifying the active lipid A moiety of endotoxin, it might be expected to protect horses after carbohydrate overload and provide a new therapeutic and experimental tool for this condition. The present study was undertaken to evaluate the effect of polymyxin B on hemostatic, hemodynamic, acid-base, and clinical aspects of the syndrome resulting from carbohydrate overload. Experimentally-induced carbohydrate overload resulted in lactic acidosis, hypercoagulability, hypovolemic shock and lameness. Although there was a slight delay in the onset of clinical signs resulting from experimental carbohydrate overload in treated animals, polymyxin B administered iv at 2.5 mg/kg every 6 hr failed to significantly ameliorate the coagulopathy, acidosis, lameness and shock induced by alimentary carbohydrate overload. PMID- 2557706 TI - Effect of paraquat on serum and lung angiotensin I converting enzyme (AICE) activity in beagle dogs. AB - The purpose of this study was to examine the relationship between lung or serum angiotensin I converting enzyme (AICE) activity and lung damage due to the administration of paraquat (PQ) which develops into PQ toxicity. Our experimental animals consisted of 19 beagle dogs (1-1.5 years old) that were allocated, randomly, into 2 treatment group, consisting of 11 PQ-treated animals and a control group of 8 saline-treated animals. These canines were administered PQ dichloride sc at a dose of 1.0 mg/kg body weight (PQ group) or sc saline at a dose of 0.5 ml/kg body weight (control group) for 7 to 11 consecutive days; these are doses which had been shown to be toxic to lungs. If a decrease in the animal's body weight was detected, the administration of PQ to that dog was discontinued. Body weight loss was observed in 10 of the 11 dogs treated with PQ. Representative dogs were sacrificed on days 11, 25 and 180 for the determination of both serum and lung AICE activity. The PQ group experienced a reduction in lung AICE activity and an elevation in the serum AICE activity. These results suggest that PQ damaged the endothelial cells of the lung and releasing lung AICE into the circulating blood, which led to an elevated total AICE activity in the serum. This pattern suggested damage to endothelial cells, indicating that PQ produced either continuing or repeated reinjury to lungs. Only the lungs were injured; therefore AICE from the rest of the body would not be expected to contribute to our AICE measurements.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557707 TI - Sepracell-MN for mononuclear cell separation from cattle: effect on mitogen-and antigen-induced lymphocyte blastogenesis. AB - Mononuclear leukocytes (MNC) were separated from heparinized and EDTA-treated whole bovine blood by centrifugation after mixing with a commercial colloidal silica preparation (Sepracell-MN (S-MN]. Cell yields and lymphocyte blast transformation (LBT) to pokeweed mitogen (PWM), phytohaemagglutinin (PHA), concanavalin A (Con A), and Brucella abortus antigens were tested against MNC obtained from heparinized whole blood using Ficoll-Hypaque (FH). Separation with S-MN was more rapid and less labor intensive than separation with FH. There was a higher average total yield of MNC but a lower percentage of monocytes in the FH- than in the S-MN-separated MNC. In mitogen-induced LBT assays, MNC responded comparably to each mitogen regardless of the separation technique or anticoagulant used, and a cell concentration effect was demonstrated. In general, FH-separated MNC responded greater to PWM than did S-MN/EDTA separated MNC, but S MN/heparin separated MNC had the greatest LBT responses to PWM. Overall, S MN/EDTA separated MNC had the greatest responses to PHA, and responses to Con A were variable among experiments with respect to the separation technique. In antigen-induced LBT assays, two B. abortus antigens were used: a heat-killed strain S1119 (HKA) and a gamma-irradiated strain 19 (gamma BA). The LBT responses of three steers vaccinated with live B. abortus strain 19 were compared with three nonvaccinated steers in three separate experiments. Using HKA, FH separation resulted in an overall greater LBT response for vaccinates than nonvaccinates and a greater differential between responses of vaccinates and nonvaccinates than did S-MN derived MNC regardless of the anticoagulant used. Using gamma BA, FH produced the most responsive MNC in one experiment and S MN/heparin produced the most responsive MNC in the other. At the highest cell concentration tested, FH-separated MNC had the greatest LBT responses for vaccinated calves, but differences between S-MN- and FH-separated MNC responses were not significantly different (P greater than 0.05). In conclusion, S-MN is a rapid and simple technique for separation of MNC from bovine blood. The technique produces an adequate cell population for mitogen-induced LBT studies; however, FH separated MNC were generally more responsive in the B. abortus-induced LBT assay. PMID- 2557708 TI - [Rehabilitative treatment in the early posthospital period of patients undergoing radical surgery of breast cancer]. PMID- 2557709 TI - [The genesis and role of fatty acids as the biologically active components of naftusia mineral water]. PMID- 2557710 TI - [Pathogenesis of the arthrosis process]. PMID- 2557711 TI - [The effect of neuropeptides on the neuronal background activity of the sensorimotor cortex]. PMID- 2557712 TI - [Effects of muscimol on aggressive behaviors induced by clonidine in mice]. AB - A behavioral study was carried out to clarify a relationship between the GABAergic and purinergic central system in aggressive behaviors induced by clonidine in mice. Mice administered a high dose of clonidine (20 mg/kg, i.p.) exhibited aggressive behaviors such as biting and attacking. These behaviors are inhibited by L-PIA (N6-L-phenylisopropyl adenosine) and stimulated by caffeine, which suggest that a blockade of adenosine receptors is involved in these behaviors. Muscimol (0.5-2 mg/kg, i.p.), a GABA-a receptor agonist, not only markedly potentiated clonidine (20 mg/kg i.p.)-induced aggressive behaviors but also elicited characteristic behaviors such as gnawing, reinforced irritability, and self-mutilation. Bicuculline (1, 2 mg/kg, i.p.), a GABA-a receptor antagonist, or picrotoxin (1 mg/kg, i.p.), a chloride channel blocker, did not significantly affect clonidine (20 mg/kg, i.p.)-induced aggressive behaviors. The potentiating effects of muscimol (0.5, 1 mg/kg, i.p.) on clonidine-induced aggressive behaviors were antagonized by bicuculline (1, 2 mg/kg, i.p.), but not affected significantly by picrotoxin (1 mg/kg, i.p.). L-PIA (0.2 mg/kg, i.p.) reduced clonidine-induced aggressive behaviors and also reversed the potentiating effects of muscimol. Stereotyped gnawing behaviors induced by combined treatment of muscimol (0.5, 1 mg/kg, i.p.) and clonidine (20 mg/kg, i.p.) were not affected by bicuculline (2 mg/kg, i.p.). The results suggest that the potentiating effects of muscimol on clonidine-induced aggressive behaviors may be induced via the stimulation of GABA-a receptors, although not necessarily associated with chloride channel functions, and may involve certain interactions between the stimulation of GABA-a receptors and the inhibition of adenosine receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557713 TI - [Diverse actions of lithium ions on transmembrane control]. PMID- 2557714 TI - [Carpal tunnel syndrome. Late results following surgical treatment]. AB - Of 64 patients, who had undergone operation because of a carpal tunnel syndrome in the years 1982 to 1984, 39 patients could be reinvestigated. The measurement of the distal motor latency was useful as well in the diagnosis as in the course of the CTS. After operation the distal motor latency was normal in about 70.7% of the cases. A subjective improvement showed 69.9% of the patients. Patients with preoperative normal motor latency reached similar postoperative results. The diagnosis cannot made only on the basis of electrophysiologic pathological values, but has to take into account the clinical picture and neurological findings. Phalen's and Tinel's signs are useful helps in the diagnosis of CTS. The results after decompression of the median nerve depend on the length of history. PMID- 2557715 TI - [Initial experiences with CA-15-3 determination in the serum of patients with breast cancer]. AB - CA 15-3 levels were determined immunoradiometrically in sera of 63 women with breast cancer (44 with stage I to III cancer, pre-operatively, 5 with local recurrence or lymphnode metastases and 14 patients with distant metastases) and 30 women with benign breast tumours or fibrocystic disease. 32% elevated levels (greater than 25 U/ml) have been found in all carcinomas compared with 7% in 44 patients without any evidence of disease (NED) and 0% in the benign lesions. In locally limited breast carcinomas, the very low pre-operative sensitivity of 16% prevents early tumour detection, whereas in patients with distant metastases a sensitivity of 86% (12 out of 14) has been found. In follow-up, continuous rising of CA 15-3 levels reveals metastatic or progressive disease with a great accuracy, decreasing serum values were only found in cases of remission. A positive correlation between the actual clinical situation in follow-up and the changes in these tumour marker levels has been observed in 87%. The CA 15-3 test system seems to be useful assay for patients with breast cancer with respect to earlier detection of distant metastases, especially of osseous type. This tumour marker is not suitable for screening or early diagnosis of small tumour recurrence in the follow-up of breast cancer patients. PMID- 2557716 TI - Radionuclide assessment of right ventricular regional wall motion abnormalities in anterior left ventricular infarction. AB - Because of its potential use in the detection of right ventricular myocardial infarction, we performed radionuclide ventriculography in 80 consecutive cases of electrocardiographically anterior acute MI. Regional wall motion of both ventricles was studied on amplitude-phase images. Forty-five patients (56%) showed normal right ventricular function and 35 (44%) regional right ventricular dyskinesia: 19 in the septal, 14 in the apical and 2 in the free wall region. Right ventricular septal, respectively apical asynergy were virtually always associated with asynergy in the homologous parts of the left ventricle. This could be a consequence of the proximity of these regions, or alternatively be due to a common vascular supply. Thus it remains uncertain if asynergy in these regions signifies necrosis of part of the right ventricular wall. Free wall asynergy was considered as evidence of right ventricular wall necrosis. Thus, in our study group the prevalence of right ventricular myocardial infarction in anterior left ventricular myocardial infarction was at least 2.5%. PMID- 2557717 TI - The somatostatin analogue SMS 201-995 in long-term treatment of vipoma. Case report. AB - In a 60-year-old woman with an inoperable vipoma, satisfactory improvement of severe watery diarrhea was obtained over a 12-month period by subcutaneous administration of the somatostatin analogue SMS 201-995. It is suggested that SMS 201-995 may be useful in such inoperable cases, or when surgery would carry a high risk. PMID- 2557718 TI - Fracture fixation with biodegradable rods. Forty-one cases of severe ankle fractures. AB - In a prospective study of 41 patients, severe ankle fractures of Lauge-Hansen types SE III-IV, PA III, and PE III-IV were treated by open reduction and internal fixation using biodegradable self-reinforced polyglycolide cylinder shaped rods. Disruption of the distal tibiofibular syndesmosis and/or fracture of the posterior tibial margin requiring reduction and fixation were the inclusion criteria for the study. The mean follow-up time after operation was 16 (12-32) months. Two failures of fixation necessitated reoperation. A secondary displacement of 1-2 mm of the lateral malleolus occurred in 3 cases. Transient accumulation of soluble polyglycolide mass complicated the course in 3 cases, but did not influence the radiographic or the functional result. Function became good in 30 patients. The advantage of the biodegradable implants is that they do not need to be removed at secondary operations. PMID- 2557719 TI - Stress shielding reduced by a silicon plate-bone interface. A canine experiment. AB - Effects of modifying the plate-bone interface with Silastic were tested. The experimental plate, i.e., a conventional plate with a Silastic backing was compared with the standard plate. In vitro four-point bending tests showed similar strain per load behavior in the experimental and standard plate models. In vitro plate-bone contact was greater and interface pressure was lower for the experimental as compared with the standard plate. An in vivo implant study was conducted where the plates were tested on intact canine femurs. At 27 weeks postimplantation, there was less porosity and remodeling in the haversian envelope of bone plated with a Silastic interface. PMID- 2557720 TI - Congenital adrenal hypoplasia and glycerol kinase deficiency. AB - An unusual case of salt-wasting in a male infant is reported. The cause was a small X-chromosomal deletion within Xp21 resulting in the syndrome of congenital adrenal hypoplasia with glycerol kinase deficiency. This syndrome can readily be diagnosed by routine biochemical tests. PMID- 2557721 TI - A prospective study of children with first acute symptomatic E. coli urinary tract infection. Early 99mtechnetium dimercaptosuccinic acid scan appearances. AB - Between 1985 and 1987 102 children, age 0-14 years, presented with a first acute symptomatic E. coli urinary tract infection. Investigations included early 99mtechnetium dimercaptosuccinic acid (DMSA) scan (which was performed at a median of 27 days), ultrasonography, micturating cysto-urethrography and indirect voiding radionuclide cystography using 99mTc DTPA. Follow-up DMSA scan was carried out after 6 months. Twenty-one of 102 of initial DMSA studies showed diminished uptake of radionuclide and 12 showed cortical scarring. Twenty-nine patients had significant vesicoureteral reflux (VUR). The finding of diminished uptake on the initial scan was significantly associated with fever, systemic upset, length of symptoms and a peripheral blood leucocytosis, (p less than 0.05). In addition the finding was associated with fever and loin pain in the older child. Both diminished uptake and scarring were more common in refluxing kidney units. We propose that, in children with UTI, diminished uptake on early DMSA scan localises infection in the renal parenchyma. PMID- 2557722 TI - Congenital cytomegalovirus infection associated with fetal ascites and intrahepatic calcifications. AB - A fetus at 20 weeks' gestation was shown by ultrasonography to have ascites and intrahepatic calcifications. We aspirated the fetal ascites at 29 and 30 weeks' gestation to decompress the fetal lungs due to the progression of the ascites and the concomitant compression in the fetal lungs. The newborn had neither hypoplasia of the lungs nor any respiratory complication, though congenital cytomegalovirus infection was present. This is the first report of such congenital cytomegalovirus infection associated with fetal ascites and intrahepatic calcifications. Careful monitoring and early intervention is necessary for a good prognosis. PMID- 2557724 TI - Lack of direct interaction of metamyzole with the brain opiate receptors of rats. AB - In earlier studies (Ivanov, Staneva-Stoycheva, 1984) we have found an inhibitory effect of metamyzole (Analgin-Pharmachim) on the contractile responses of electrically stimulated guinea-pig ileum and mouse vas deferens, which is antagonized by naloxone. In the present study an attempt is made to clarify whether these presynaptic effects of metamyzole are due to direct interaction with presynaptic opiate receptors. The method of radioligand binding of /3H/ naloxone to the opiate receptors in a crude membrane fraction of rat brain was used. It was found that in the wide concentration range used (10(-5)-10(-9) M), metamyzole does not compete with /3H/-naloxone for binding to the receptor sites. The biochemical study has shown that the presynaptic opiate receptors do not participate in the mechanism of the presynaptic effects of metamyzole, established in earlier studies. PMID- 2557723 TI - Oxytocin regulates Ca2+ level in myometrium by influencing phosphoinositide metabolism. AB - The effect of oxytocin on phosphoinositide metabolism as well as on membrane protein phosphorylation in myometrial tissue was studied. Oxytocin enhanced the 32P incorporation into phospholipids in myometrial tissue. The effect of oxytocin on phosphoinositide metabolism was also detected in plasma membrane of 20 days pregnant rats. Phosphorylated membrane lipids have been analysed and phosphatidylinositol 4, 5-bisphosphate proved to be the main reaction product. Oxytocin enhanced the 32P incorporation into phospholipids measured in the first 30 sec then the labeling decreased more rapidly then in case of the control. The effect of oxytocin proved to be concentration dependent. The protein phosphorylation was also influenced by oxytocin. However the amount of alkylphosphate formed depended on the presence or absence of Ca2+, Ca2+ calmodulin and cyclic AMP, oxytocin influenced the protein phosphorylation in the presence of Ca2+-calmodulin only. PMID- 2557725 TI - Counter current transfer in the female adnex. AB - The utero-ovarian veins and lymph vessels are intimately connected with the ovarian artery in the human female and in domestic animals, with the exception of the horse and the human female. A direct, local exchange of molecules from veins and lymph vessels to arteries (counter current transfer) has been documented for this anatomic structure. Countercurrent transfer of certain inert gases (133xenon, 85krypton), of prostaglandins (PGF2 alpha), of steroid hormones (e.g. progesterone, estradiol, testosterone), and of small peptide hormones (oxytocin, relaxin) has been shown to occur in laboratory and domestic animals as well as in the human female. The transfer of the inert gases takes place within seconds. The transfer of steroid hormones and peptides is detectable within minutes while the transfer of PGF2 alpha is delayed for 20 minutes. Red blood cells or albumin are not transferred. The existence of the local transfer is postulated to be of importance for: 1) the pregnancy/non-pregnancy signal from the uterus and tube to the ovary. The signal may be a combination of a luteotrophic signal from the embryo and lack of a "non-pregnant" luteolytic signal from the endometrium, the latter probably being PGF2 alpha in some species; 2) the unilateral influence of the ovarian hormones on the function of the ovarian, tubal, and possibly uterine tissues. An active corpus luteum may create in a mono-ovulatory animal a higher progesterone level in arterial blood supplying the ipsilateral tube and ovarian interstitial tissue than on equivalent contralateral organs. PMID- 2557726 TI - Modulatory interactions of neuropeptide Y (NPY) on sympathetic neurotransmission. AB - I. Based on experiments on the rat portal vein, it is suggested that NPY is capable of inhibiting TNS-induced release of 3H-NA from the sympathetic nerves via a prejunctional site of action, and that NPY enhances the spontaneous contractility and the vasoconstrictor response to TNS via a postjunctionally mediated mechanism. II. Based on experiments in the pithed rat, it is suggested that NPY inhibits the PNS-induced enhancement of plasma NA and A levels, and that this reflects an inhibitory effect of NPY on the release of NA from sympathetic nerve terminals and the secretion of A from the adrenal medulla. III. Based on experiments in the pithed rat, it is suggested that infusion of NPY, at subthreshold doses, potentiates the blood-pressure response to PNS and to injection of the alpha-agonist phenylephrine and that NPY in higher doses induces pressor responses, which are resistant to alpha- and beta-adrenoceptor blockade but sensitive to the calcium antagonist nifedipine. IV. Based on experiments in the pithed rat, it is suggested that A is secreted from the adrenal medulla directly into the circulating blood, since the peak plasma A level occurred immediately (0-20 s) after the initiation of PNS, and that NA and NPY are released from sympathetic nerve terminals from where the transmitters have to diffuse into the systemic circulation, since the maximal levels of circulating NA and NPY-LI were reached later (20-40 s) than A. V. Based on experiments in the pithed guinea-pig, it is suggested that the PNS-induced increase of the plasma NPY-LI level is modulated by a prejunctional alpha 2-adrenoceptor mediated inhibitory control mechanism, since the alpha 2-agonist clonidine was found to reduce and the alpha 2-antagonist yohimbine to markedly enhance the PNS-induced increase in plasma NPY-LI, whereas the alpha 1-antagonist prazosin had no effect. Furthermore, guanethidine reduced this parameter, lending further support to the assumption that circulating NPY is of sympathetic neuronal origin. VI. Based on experiments in the pithed guinea-pig, it is suggested that prejunctional facilitatory beta-adrenoceptors may also be involved in the regulation of the NPY release, since infusion of isoprenaline elicited a facilitation of the PNS induced increase of plasma NPY-LI levels which could be antagonised by propranolol.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557727 TI - Alkaline secretion by Necturus proximal duodenal mucosa. AB - Proximal duodenum from the amphibian Necturus was stripped of muscle layers and the mucosa was mounted as a tube for studies of alkali transport or as a flat sheet for intracellular impalement by voltage-sensitive glass micro-electrodes. The mucosa alkalinized the unbuffered luminal perfusate at a high rate (3.4 muequiv. cm-1 h-1) and developed a transepithelial electric potential difference of 5.7 mV (lumen negative). Transport was inhibited by 2,4-dinitrophenol (10(-4) M) and by furosemide (10(-3) M) and SITS (10(-3) M) on the seros but not on the mucosal side, indicating dependence on tissue metabolism and on serosal membrane Cl-/HCO3- exchange. Prostaglandin E2 (10(-7)-10(-5) M) and dibutyryl cyclic AMP (10(-6)-10(-4) M) had no effects on the secretion or transepithelial electrical potential difference. removal of serosal HCO3- decreased luminal alkalinization by 75%, indicating a contribution by passive migration of HCO3- and/or a dependence of transcellular transport on the nutrient supply of this ion. Administration of HCO3- (17.8 mM) to the luminal perfusate affected neither the transepithelial nor transmembrane electrical potential differences nor the resistance ratio. It is thus unlikely that the luminal membrane possesses any major HCO3- conductance. PMID- 2557728 TI - Characterization of the actions of botulinum neurotoxin type E at the rat neuromuscular junction. AB - Botulinum neurotoxin (BoTx) serotype E blocks spontaneous and evoked quantal release of acetylcholine at the rat neuromuscular junction. Increasing extracellular Ca2+ to 8 mmol l-1 or substituting Ca2+ with La3+ (0.1 and 1.0 mmol l-1) or depolarizing the nerve terminals by 20 mmol l-1 K+ markedly increases miniature end-plate potential frequency in normal muscle, but in BoTx-E poisoned preparations none of these ions, with the exception of 1 mmol l-1 La3+, was able to restore spontaneous quantal transmitter release to levels recorded at unpoisoned junctions. In absolute values the enhancement with La3+ was much less than that reported at normal junctions. Nerve stimulation in the presence of 3,4 diaminopyridine (10-20 mumol l-1) and high calcium (8 mmol l-1) evoked multiquantal end-plate potentials and muscle twitches. We conclude that the neuromuscular block produced by BoTx serotype E is similar to that previously described for BoTx serotype A but differs from that produced by BoTx serotypes B, D and F in not causing desynchronization of nerve impulse-evoked transmitter release. 3,4-Diaminopyridine might be useful in the treatment of poisoning by BoTx serotype E since it markedly enhanced synchronous transmitter release from poisoned motor nerve terminals. PMID- 2557729 TI - Perchlorate stimulates potassium-induced growth hormone release in cultured rat somatotrophs. PMID- 2557730 TI - Neurokinin A enhances the stimulatory effects of d-amphetamine on motor activity in the nucleus accumbens of the rat. PMID- 2557731 TI - Localization of [125I]endothelin-1 and [125I]endothelin-3 binding sites in the rat brain. PMID- 2557732 TI - Lipoxin A4 inhibits leukotriene B4-induced inflammation in the hamster cheek pouch. PMID- 2557733 TI - The trophic responses of avian sensory ganglia in vitro to N-acetylated and des acetyl forms of alpha-melanocyte stimulating hormone (alpha-MSH) are qualitatively distinct. AB - alpha-Melanocyte-stimulating hormone (alpha-MSH) accelerates the regrowth of peripheral nerve axons in the rat following their transection (Verhaagen et al., Expl Neurol. 92, 451-454, 1986). The cellular mechanisms of this trophic response were investigated for several naturally occurring derivatives of alpha-MSH using Nerve Growth Factor (NGF)-stimulated quail sensory ganglion explants in vitro in which both neurite outgrowth and non-neuronal cell behaviour could be more reliably observed and quantified. Neurite outgrowth was determined with a semi quantitative scoring assay. Glial migration into the outgrowth was quantified using a monoclonal antibody, GTE-52, which labels the nuclei of Schwann cells. Des-acetyl alpha-MSH caused a marginal increase in the neurite outgrowth density which was significant at concentrations of 0.04 and 0.1 microgram/ml. The response to acetylated (N-acetyl, N,O-diacetyl) forms of alpha-MSH was characterized by fascicle formation by neurites which resulted in an apparent decrease in the neurite score, and by the outgrowth of non-neuronal cells. Using monoclonal antibody GTE-52, which recognizes a glial nuclear antigen, these cells were identified as Schwann cells. N-Acetyl, but not des-acetyl alpha-MSH increased the number of GTE-52-labelled cells in the NGF-stimulated neurite outgrowth and stimulated their migration in the absence of neurites when NGF was omitted from the culture medium. Exposure of growing explants to two polyclonal antibodies against alpha-MSH resulted in an increased neurite outgrowth density. The results support the hypothesis that alpha-MSH peptides stimulate peripheral nerve growth by modulating the neurite sprouting response, and demonstrate that the nature of the neurotrophic response to naturally occurring melanotropins depends on the existence of acyl substitution at the N-terminal amino acid residue. A possible role of endogenous melanotropin peptides in the regulation of sensory nerve growth is discussed. PMID- 2557735 TI - Intramedullary spinal cord metastasis detected through intrathecal contrast medium computerized tomography. AB - A case of intramedullary spinal cord metastasis is reported in which the diagnosis was possible through intrathecal contrast medium computerized tomography. The case described suggests that this method of radiologic investigation can provide a prompt identification of ISM from other noncompressive complications of systemic cancers. This is very important for radiation treatment which can be effective in the early stages of ISM. PMID- 2557734 TI - L-acetylcarnitine attenuates the age-dependent decrease of NMDA-sensitive glutamate receptors in rat hippocampus. AB - NMDA-sensitive glutamate receptors are involved in the regulation of neuronal plasticity, and contribute to the synaptic mechanisms underlying the learning process. Aging is associated with a reduction in the maximal density of NMDA sensitive glutamate binding sites in rat hippocampus. This reduction is attenuated after long-term administration with L-acetylcarnitine (10 mg/Kg i.p. once a day for 4 months). These results support a neuroprotective and neurotrophic role for L-acetylcarnitine during aging. PMID- 2557736 TI - Regional distribution of angiotensin converting enzyme in the rat kidney. PMID- 2557737 TI - Immunohistologic characterization of angiotensin converting enzyme in the human kidney using monoclonal and polyclonal antibodies. PMID- 2557738 TI - Effects of mineralocorticoid on kininase activity along the distal nephron segments of the rat. PMID- 2557739 TI - Selective expression of des-Arg9-BK sensitive (B1) receptors in vivo and in vitro by angiotensin converting enzyme inhibitors. AB - LPS (E. Coli), captopril, enalapril, teprotide and BK-infusion induced after 18 h in rabbits an increased hypotensive response to the selective B1-receptor agonist des-Arg9-BK. A similar selective increase in responsiveness and sensitivity of the in vitro aortic strip to des-Arg9-BK taken from pretreated animals was also seen. These results suggest that angiotensin converting enzyme inhibitors may elevate endogenous kinin levels which may contribute towards the inflammatory side-effects seen with this group of drugs. PMID- 2557740 TI - Demonstration of in vivo effects of ACE inhibitors by the use of autoregressive modelling. AB - We investigated, with use of a technique of feedback analysis, the dynamic changes in and around the renin-angiotensin system in vivo induced by ACE inhibition. Prolonged studies were performed over a half year on rabbits, which were treated by daily injections of saline or one of the inhibitors as follows: captopril, foroxymithine, histargin and pepstatin. In spite of the apparently bizarre movements of the raw data, the analysis with autoregressive modelling unveiled the different modes of feedback regulations of the renin-angiotensin system under the influence of the various inhibitors. In order to interpret the in vivo actions of enzyme inhibitors, it seems essential to take feedback regulations into consideration. PMID- 2557741 TI - Increased insulin-responsiveness by ACE-inhibition in noninsulin dependent diabetes mellitus. PMID- 2557742 TI - Angiotensin I converting enzyme. PMID- 2557743 TI - Relationship between ANP, cyclic GMP and tissue kallikrein following saline infusion in healthy volunteers. AB - Infusion of saline in healthy volunteers produced a significant rise in plasma atrial natriuretic peptide, which was accompanied by the urinary excretion of cyclic GMP and sodium. Although the time-course of cyclic GMP excretion almost mirrored that of plasma ANP, that of sodium showed sustained phase, indicating involvement of additional humoral regulators. ANP did not stimulate the excretion of enzymic or immunoreactive tissue kallikrein. PMID- 2557744 TI - Pharmacokinetics of angiotensin converting enzyme inhibitors in plasma and tissue using radioinhibitor binding and displacement assays. PMID- 2557745 TI - Kininase activities in the human pituitary gland. PMID- 2557746 TI - Purification of human seminal plasma kininase II using affinity chromatography. PMID- 2557747 TI - The mechanism of degradation of bradykinin (lysyl-bradykinin) in human serum. PMID- 2557748 TI - Role of the endogenous angiotensin II in the antihypertensive effect of MK 421 in rats made hypertensive by norepinephrine or vasopressin. AB - To assess the mechanism by which inhibitors of angiotensin converting enzyme (ACE) lower blood pressure, we evaluated the role of endogenous angiotensin II in the antihypertensive effect of MK 421, a long-lasting ACE inhibitor, in rats made hypertensive by chronic infusion of norepinephrine or vasopressin. The hypertensive effect of norepinephrine (1.8 mg/kg/day, ip) or vasopressin (7.2 U/kg/day, ip) was inhibited by the simultaneous administration of MK 421 (6 mg/kg/day, ip). Additional administration of angiotensin II at a subpressor dose (36 micrograms/kg/day, ip) did not revert the antihypertensive effect of MK 421 in rats made hypertensive by chronic infusion of norepinephrine or vasopressin. The present results suggest that the hypotensive effect of ACE inhibitors may depend on a reduced sensitivity of the vasculature to vasoconstrictor substances. In addition, it is also suggested that the suppressed angiotensin II may not be essential for the antihypertensive effect of ACE inhibitors in rats made hypertensive by chronic infusion of norepinephrine or vasopressin. PMID- 2557749 TI - An iodinatable photoaffinity probe based on the structure of kallidin. PMID- 2557750 TI - Specific binding sites for bradykinin and its degradation process in cultured rat vascular smooth muscle cells. PMID- 2557751 TI - Kinin receptors on epithelial cells and smooth muscle of the trachea. AB - Kallidin and bradykinin produce a biphasic response, contraction followed by relaxation on the isolated guinea-pig tracheal muscle. Two population of receptors appear to regulate the action of kinins on the trachea. Receptors on the epithelium seem to release chemical mediators that control the relaxation response. The question whether similar or different receptors modulate contraction of the smooth muscle may be answered by the use of appropriate antagonists. The occurrence of kinins in bronchial lavage of patients with asthma raises the question of the source of tissue kallikrein involved in the formation of kallidin. We have identified immunoreactive tissue kallikrein in the submucous glands of the trachea. PMID- 2557752 TI - Human urinary kallikreins A, B and D: isolation, purification and properties. PMID- 2557753 TI - Tissue kallikrein effectively activates latent matrix degrading metalloenzymes. PMID- 2557754 TI - Therapeutic regime of enalapril maleate in Japan. PMID- 2557755 TI - Tonin and kallikrein-kinin system. AB - The kininogenase activity of tonin has been demonstrated by Ikeda and Arakawa, 1984. Tonin of the rat submandibular gland contracts the rat uterus independent of addition of the substrate. On repetition, the same dose of enzyme elicited desensitization. When a double dose was used the contraction again occurred. After desensitization to tonin the contraction to kallikrein was reduced about 80% of the control. The desensitization to kallikrein lightly reduced the contraction to tonin. When the muscle was desensitized to trypsin tonin did not evoke contraction. These experiments suggest the presence of two different substrates in the uterus, one more specific to kallikrein and the other for tonin. The experiments with the parallel uterus preparation strongly suggest release of kinin in the process of contraction of the uterus by tonin. PMID- 2557756 TI - Serum kininases I and II in patients with extrinsic and intrinsic asthma. PMID- 2557757 TI - Physiological role of renal kallikrein-kinin system in human. AB - The physiological role of renal kallikrein-kinin system in human is discussed by the three following topics. Localization of each component of renal kallikrein kinin system: Kallikrein is localized in the apical site of the distal tubular epithelial cells and collecting ducts by the immunostaining method. Following the stop-flow method in dog kidney, kallikrein and kinin are recognized in the distal tubules. Kininase I, II and neutral endopeptidase (enkephalinase) are localized not only in the proximal tubules, but also in the distal tubules. The localization of kininases is further confirmed by the stop-flow method pretreated with specific inhibitors for each of the kininases. Sodium metabolism and renal kallikrein-kinin system: In normal subjects, fractional excretions of sodium and inorganic phosphorus which reflect the total and proximal sodium reabsorption, show significantly positive correlations for both urinary kallikrein and kinin excretions. In the case of 0.9% saline infusion, the activity of renal kallikrein kinin system is augmented following the infusion as shown in the increases of urinary kallikrein and kinin excretions and the decreases of urinary kininases excretions. Relation to other renal depressor systems: The close relations among renal dopamine, kallikrein-kinin and prostaglandin systems have been suggested by a dopamine infusion study. Dopamine may augment the activity of the renal kallikrein-kinin system through both the increases of renal prekallikrein synthesis and kallikrein specific activity. From these studies reported up to the present, it is suggested that the renal kallikrein-kinin system produced in the distal nephron in the kidney may play a role in the sodium metabolism with other renal depressor systems in addition to its own action. PMID- 2557760 TI - The role of viral and cellular nuclear proteins in herpes simplex virus replication. AB - Following infection of cells by herpes simplex virus, the cell nucleus is subverted for transcription and replication of the viral genome and assembly of progeny nucleocapsids. The transition from host to viral transcription involves viral proteins that influence the ability of the cellular RNA polymerase II to transcribe a series of viral genes. The regulation of RNA polymerase II activity by viral gene products seems to occur by several different mechanisms: (1) viral proteins complex with cellular proteins and alter their transcription-promoting activity (e.g., alpha TIF), (2) viral proteins bind to specific DNA sequences and alter transcription (e.g., ICP4), and (3) viral proteins affect the posttranslational modification of viral or cellular transcriptional regulatory proteins (e.g., possibly ICP27). Thus, HSV may utilize several different approaches to influence the ability of host-cell RNA polymerase II to transcribe viral genes. Although it is known that viral transcription uses the host-cell polymerase II, it is not known whether viral infection causes a change in the structural elements of the nucleus that promote transcription. In contrast, HSV encodes a new DNA polymerase and accessory proteins that complex with and reorganize cellular proteins to form new structures where viral DNA replication takes place. HSV may encode a large number of DNA replication proteins, including a new polymerase, because it replicates in resting cells where these cellular gene products would never be expressed. However, it imitates the host cell in that it localizes viral DNA replication proteins to discrete compartments of the nucleus where viral DNA synthesis takes place. Furthermore, there is evidence that at least one specific viral gene protein can play a role in organizing the assembly of the DNA replication structures. Further work in this system may determine whether assembly of these structures is essential for efficient viral DNA replication and if so, why assembly of these structures is necessary. Thus, the study of the localization and assembly of HSV DNA replication proteins provides a system to examine the mechanisms involved in morphogenesis of the cell nucleus. Therefore, several critical principles are apparent from these discussions of the metabolism of HSV transcription and DNA replication. First, there are many ways in which the activity of RNA polymerase II can be regulated, and HSV proteins exploit several of these in controlling the transcription of a single DNA molecule. Second, the interplay of these multiple regulatory pathways is likely to control the progress of the lytic cycle and may play a role in determining the lytic versus latent infection decision.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557759 TI - The outbreak of poliomyelitis in Finland in 1984-1985: significance of antigenic variation of type 3 polioviruses and site specificity of antibody responses in antipolio immunizations. PMID- 2557758 TI - Human papillomaviruses and carcinomas. AB - The recognition of multiple types of human papillomaviruses has resulted in remarkable progress in the detection of persisting viral nucleic acid sequences in carcinomas. The consistent transcription in tumors of two early open reading frames, E6 and E7, with few exceptions (Lehn et al., 1985), indicates a role for the products of these genes in the induction and/or maintenance of the transformed state. A number of studies have shown that in vitro transformation can be achieved by transfection of E6/E7 DNA, and proteins encoded by these DNA sequences can be demonstrated in primary human keratinocytes immortalized by this DNA (Kaur et al., 1989). Mutagenesis experiments are needed to determine the absolute requirement for and function of these genes in transformation. A preferential association of some types with benign lesions while others may be frequently found in malignant tumors has been observed. HPV types 5 and 8 in epidermodysplasia verruciformis patients and types 16, 18, 31, 33, etc. in genital lesions are most frequently associated with progression to malignancy, whereas other types, such as HPV-6,-10, -11, and -20, are regularly identified in benign warts. Such distinctions are not absolute but provide the initial steps toward establishing a causal role for some human papillomaviruses in carcinomas. The need for well-designed epidemiological studies in concert with optimum molecular and serologic evaluations is evident (Armstrong et al., 1988). The data from human and animal studies indicate that papillomaviruses contribute significantly to the development of many, if not all, carcinomas, but we do not yet have a clear understanding of the importance of other interacting viral, chemical, or cellular factors. The application of gene cloning and non-stringent hybridization (Law et al., 1979) has provided us with an apparently ever increasing catalog of human papillomaviruses. More effort is now required to establish their prevalence, the natural history of infection, and the mechanism of neoplastic transformation. PMID- 2557762 TI - Chemical evolution of the fluoroquinolone antimicrobial agents. AB - In the past decade, significant progress has been made in understanding structure function relationships of the new quinolones, which have a N-1-substituted, 1,4 dihydro-4-oxo-pyridine-3-carboxylic acid moiety as the basic nucleus. Modification of the groups affixed to positions C-6, C-7, and C-8 has made a major change in the antimicrobial activity, pharmacokinetic, and metabolic properties of the quinolones as have changes in the moieties affixed to the N-1 nitrogen. The new quinolones have a carboxyl group at position 3 and a keto group at C-4. The presence of a fluorine atom at C-6 enhances the deoxyribonucleic acid (DNA) gyrase inhibitory activity as well as the ability of the compounds to inhibit staphylococci. Position C-7 has been one of the most modified sites. Addition of a piperazinyl group markedly increased gram-positive activity, primarily antistaphylococcal activity; lowered the minimal inhibitory concentrations against Enterobacteriaceae, Haemophilus spp., and Neisseria spp.; and added activity against Pseudomonas aeruginosa compared with nalidixic acid. Methyl derivatives of the piperazine group or of the pyrroles have longer half lives than do unsubstituted moieties. At the N-1 position, a cyclopropyl group appears to be most potent with respect to minimal inhibitory concentrations against Enterobacteriaceae and Pseudomonas. Ofloxacin is unique in that it has an oxygen substituted at C-8 with the substituent part of the ring system formed by fusion to the N-1 position. This has produced excellent in vitro activity against gram-positive species comparable with that of ciprofloxacin, excellent activity against the Enterobacteriaceae, and antipseudomonal activity superior to agents with an ethyl substitution at position N-1. The oxazine ring of ofloxacin provides excellent oral absorption with virtually 95 percent bioavailability; this modification also has prevented metabolism and has provided a long half-life of seven to eight hours. PMID- 2557761 TI - A single measurement of adrenocorticotrophic hormone in cerebrospinal fluid is of no value in the diagnosis of dementia. AB - It has been reported that adrenocorticotrophic hormone (ACTH) levels in the cerebrospinal fluid (CSF) of dementia patients are lower than in normal subjects. We have carried out ACTH estimations in the CSF of 17 patients suffering from dementia and on sex- and age-matched controls. No difference was found. PMID- 2557763 TI - Vasopressin receptors in human pregnant myometrium and decidua: interactions with oxytocin and vasopressin agonists and antagonists. AB - Saturation analysis and competition experiments were performed to identify and characterize [3H]arginine vasopressin binding to human myometrium and decidua in late pregnancy. [3H]Arginine vasopressin bound with affinity similar to that of [3H]oxytocin to both tissues (dissociation constant 1 to 2 nmol/L). The concentration of [3H]arginine vasopressin binding sites was high, particularly in decidua, but in all instances was about 50% to 60% of [3H]oxytocin binding. Analogs with selective oxytocic potency (4-threonine oxytocin, isotocin) had high affinity to both [3H]arginine vasopressin and [3H]oxytocin binding sites, as did analogs with both oxytocic and vasopressor activity (vasotocin). Analogs with selective antidiuretic activity (1-deamino-8-D-arginine vasopressin) showed drastically reduced affinity to [3H]oxytocin binding sites and relatively low but significantly higher affinity to [3H]arginine vasopressin binding sites. A new oxytocin antagonist (RW22164 or [1-deamino-2D-tyrosine-(O-ethyl)-4-threonine-8 ornithine]oxytocin) competitively bound to both binding sites. Its affinity to [3H]oxytocin binding sites was greater than to [3H]arginine vasopressin binding sites whereas the relative affinities of a predominantly vasopressor antagonist [Manning compound) were reversed, suggesting the presence of distinct receptors for oxytocin and arginine vasopressin in pregnant human myometrium and decidua. PMID- 2557765 TI - Increased cAMP in proximal tubules is acute effect of nicotinamide analogues. AB - The possible role of adenosine 3',5'-cyclic monophosphate (cAMP) in the mechanism of the acute inhibitory effects of nicotinamide and analogues on brush-border membrane (BBM) phosphate transport was investigated. Compared with basal values, cAMP content of rat renal proximal tubule suspensions was elevated two- to fivefold when incubated at 37 degrees C for 1 h with nicotinamide, 5 methylnicotinamide, or picolinamide at 1-3 mM and in the presence of a phosphodiesterase inhibitor. Thymidine had no effect on cAMP content. There was significant and specific inhibition of BBM transport of phosphate when proximal tubules were incubated with either nicotinamide or picolinamide at concentrations that increased tubule cAMP content. Thymidine had no effect on BBM transport of phosphate. These findings were independent of the dietary Pi intake of the rats. The absence of any effect of thymidine on phosphate transport strongly suggests that inhibition of poly(adenosine diphosphate ribose) polymerase does not play a role in nicotinamide action on phosphate transport. The change in phosphate transport induced by nicotinamide occurred with no change in NAD content. These findings indicate that an increase in cAMP, rather than NAD, is the important change that may mediate the acute inhibition of Na(+)-dependent phosphate transport by nicotinamide. PMID- 2557764 TI - Comparison of Cytobrush and cervicovaginal lavage sampling methods for the detection of genital human papillomavirus. AB - The development of an accurate method for the detection and typing of genital human papillomavirus is of substantial clinical importance. This virus has been implicated as an etiologic agent in the development of cervical neoplasia. To detect human papillomavirus infection with maximum sensitivity, cells must be collected and assayed for human papillomavirus deoxyribonucleic acid. We compared two noninvasive methods of sampling exfoliated cervical cells--cervicovaginal lavage and scrape-Cytobrush. Seventy-four patients newly referred to the colposcopy clinic were divided randomly for cell sampling by either cervicovaginal lavage followed by scrape-Cytobrush or, conversely, scrape Cytobrush followed by cervicovaginal lavage. Restriction analysis and Southern blot hybridization were used to test all the samples thus obtained for human papillomavirus. Overall, test results from 42 patients (56.8%) were positive for human papillomavirus deoxyribonucleic acid. Twenty-six (31.1%) tested positive for human papillomavirus by both sampling methods, and 32 (43.2%) tested negative for human papillomavirus by both methods. One (1.4%) tested positive with scrape Cytobrush sampling but negative with cervicovaginal lavage, while 15 (20.3%) tested negative with scrape-Cytobrush but positive with cervicovaginal lavage (p less than 0.001, McNemar's test). These data, combined with previous work from our group, suggest that, of the available methods, cervicovaginal lavage, coupled with human papillomavirus deoxyribonucleic acid hybridization, is the most sensitive noninvasive method for harvesting cells for molecular identification of human papillomavirus in the female lower genital tract. PMID- 2557766 TI - Sodium-phosphate cotransport in OK cells: inhibition by PTH and "adaptation" to low phosphate. AB - Sodium-phosphate (Na-Pi) cotransport is principally regulated by parathyroid hormone (PTH) and by the intrinsic ability to "adapt" to ambient phosphate concentration. In the present study, these two control mechanisms were examined in a cloned opossum kidney (OK) cell line. PTH inhibited Na-Pi cotransport, half maximal inhibition at 5 x 10(-12) M, by fractionally similar amounts irrespective of the initial transport rates predetermined by "adaptation" to media phosphate concentration. At maximal concentrations of PTH (10(-8) M), the residual Na-Pi cotransport activity was higher in cells exposed to low-phosphate media. Cells preexposed to PTH (10(-8) M) or dibutyryl adenosine 3'-5'-cyclic monophosphate (DBcAMP) (10(-5) M) and forskolin (10(-5) M) increase transport (adaptation) by fractionally similar amounts as control cells for any given external phosphate concentration. The protein kinase C inhibitor, staurosporine, prevented PTH action but did not alter the ability to adapt Na-Pi cotransport in response to low media phosphate concentration. These data support the notion that regulation of Na-Pi cotransport by PTH and the adaptive response to available media phosphate concentration are distinct regulatory control mechanisms. PMID- 2557767 TI - Y2-type receptors for peptide YY on renal proximal tubular cells in the rabbit. AB - By means of primary cell cultures and luminal and basolateral membrane vesicles a single class of high-affinity binding sites for peptide YY (PYY), a member of the pancreatic polypeptide (PP)-fold family of peptides, was identified on the vascular side of the tubular epithelium in the proximal convoluted tubule of rabbit kidney. The binding of mono-iodinated radiolabeled PYY was inhibited equally well by PYY and neuropeptide Y (NPY), whereas the potency of the third member of the family, PP, was 10(5) times lower. Because NPY immunoreactive nerves in the mammalian kidney are confined to vascular smooth muscle cells and the juxtaglomerular apparatus, we propose that the physiological ligand for this binding site is blood-borne PYY. The kidney PYY receptor was sensitive to guanine nucleotides and could be classified as belonging to the Y2-subtype of NPY receptors, thus resembling in its binding characteristics the hippocampal NPY receptor. The high amounts of Y2-type PYY receptors present on the proximal tubule cell in rabbit kidney should permit studies on the functions and mechanisms of actions of PYY. PMID- 2557768 TI - Modulation of ionic permeability in a nonpolarized cell: effect of cAMP. AB - To evaluate whether adenosine 3',5'-cyclic monophosphate (cAMP) modulates ionic permeabilities of nonpolarized cells, as reported in diverse polarized epithelia, relative ionic permeabilities were determined in human red cell ghosts by means of the potential-sensitive fluorescent probe 3,3'-dipropylthiadicarbocyanine iodide. Relative ionic chloride permeability (PCl/PK), but not PNa/PK, was significantly increased in ghosts prepared from normal red blood cells (RBCs) exposed to cAMP analogues or forskolin, with the latter at a concentration that significantly increased intracellular cAMP concentration. As basal RBC cAMP concentrations of untreated uremic subjects were also increased, relative permeabilities of ghosts and unstimulated RBC cAMP concentrations were compared in normal, uremic, and dialyzed subjects, PCl/PK was significantly increased in uremic compared with normal subjects; PNa/PK was not altered. PCl/PK and RBC cAMP concentrations were indistinguishable in normal and dialyzed subjects. Neither the kinetics nor number of Cl(-)-HCO3- antiporters, assessed with the pH sensitive probe acridine orange and the disulfonic stilbene 4,4' diisothiocyanostilbene-2,2'-disulfonic acid, respectively, were altered in uremic cells. These studies suggest that cAMP modulates ionic chloride permeability via increased chloride conductance of each Cl(-)-HCO3- antiporter or by activation/opening of new or existing channels in RBC membranes. PMID- 2557769 TI - Contributions of a transient outward current to repolarization in human atrium. AB - Conventional microelectrode recordings combined with enzymatic cell dispersion methods and a single microelectrode voltage-clamp technique were used to record transmembrane action potentials and ionic currents in isolated single myocytes and in excised segments of human right atrium. Recordings of the outward current(s), which is responsible for the resting potential and early repolarization of the action potential in human right atrium, consistently showed that this tissue has 1) a relatively small inwardly rectifying background potassium current (IK1) which generates the resting potential in mammalian ventricular tissue and Purkinje fibers, and 2) a large time- and voltage dependent, but Ca2(+)-independent, transient outward current. A somewhat similar K+ current was originally described in neurons and recently has also been identified in a variety of mammalian cardiac tissues. As expected from previous work, this transient outward current in human atrium is blocked by 4 aminopyridine (4-AP; 0.5 mM) and exhibits time- and voltage-dependent inactivation and reactivation. Measurements of action potential shape changes and phasic tension as a function of stimulus frequency, or after 4-AP application, show that in human atrium this current can produce pronounced changes in both the early repolarization of the action potential and force generation. PMID- 2557770 TI - Role of xanthine oxidase in postischemic microvascular injury in skeletal muscle. AB - Previous reports indicate that allopurinol, a xanthine oxidase inhibitor, attenuates the microvascular injury produced by reperfusion of ischemic skeletal muscle. To further assess the role of xanthine oxidase in ischemia/reperfusion (I/R) injury, we examined the effect of xanthine oxidase depletion or inhibition on the increase in microvascular permeability produced by I/R. Changes in vascular permeability were assessed by measurement of the solvent drag reflection coefficient for total plasma proteins (sigma) in rat hindquarters subjected to 2 h of ischemia and 30 min of reperfusion in xanthine oxidase-replete and -depleted animals and in animals pretreated with the xanthine oxidase inhibitor oxypurinol. Xanthine oxidase depletion was accomplished by administration of a tungsten supplemented (0.7 g/kg diet), molybdenum-deficient diet. In animals fed the tungsten diet, muscle total xanthine dehydrogenase plus xanthine oxidase activity was decreased to less than 10% of control values. Estimates of sigma averaged 0.85 +/- 0.04 in nonischemic (continuous perfusion for 2.5 h) hindquarters, whereas muscle xanthine oxidase activity averaged 3.3 +/- 0.4 mU/g wet wt. I/R was associated with a marked decrease in sigma (0.54 +/- 0.02), whereas xanthine oxidase activity was increased to 5.8 +/- 0.5 mU/g wet wt. These results indicate that I/R produced a dramatic increase in vascular permeability coincident with an increase in muscle xanthine oxidase activity. Xanthine oxidase depletion with the tungsten diet or pretreatment with oxypurinol attenuated this permeability increase (sigma = 0.72 +/- 0.03 and 0.77 +/- 0.7, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557771 TI - Adenosine stimulates glycolytic flux in isolated perfused rat hearts by A1 adenosine receptors. AB - This study was designed to assess the role of adenosine in the regulation of exogenous glucose utilization by myocardium. Perfusion of isolated rat hearts with buffer containing D-[3-3H]glucose and analysis of the coronary effluent for 3H2O production was used as an indicator of glycolytic flux. Initially, glycolytic flux was determined during five different conditions: 1) normoxia; 2) normoxia plus 100 microM adenosine; 3) normoxia plus 100 microM adenosine and 10 microM 8-(sulfophenyl)-theophylline (SPT), an adenosine receptor antagonist; 4) hypoxia; and 5) hypoxia plus 10 microM SPT. Both adenosine and hypoxia produced an approximate threefold increase in glycolytic flux that was attenuated by adenosine receptor blockade with SPT. Next, hearts were perfused during normoxic conditions with various concentrations of either R-phenylisopropyladenosine (PIA), an A1-adenosine receptor agonist, or 5'-N-ethylcarboxamidoadenosine (NECA), an A2-adenosine receptor agonist. Significant increases in glycolytic flux occurred with PIA, whereas NECA treatment resulted in only a marginal stimulation of glycolytic flux. These data provide evidence that: 1) exogenous adenosine stimulated glycolytic flux in the normoxic myocardium; 2) endogenous adenosine stimulated glycolytic flux during hypoxia; and 3) the effect of adenosine on glycolytic flux was mediated by interaction with A1-adenosine receptors. PMID- 2557772 TI - Hydrolysis of a synthetic angiotensin-converting enzyme substrate in dog lungs. AB - The present study was carried out to begin to evaluate the saturable kinetics of the hydrolysis of a synthetic substrate, benzoyl-phenylalanyl-alanyl-proline (BPAP), for angiotensin-converting enzyme (ACE), by the pulmonary endothelium of the dog using a multiple indicator dilution method. In the experiments, isolated dog lung lobes were perfused with a salt solution containing 5% bovine serum albumin. Boluses containing [3H]BPAP, and various amounts of unlabeled BPAP were injected into the lobar artery, and timed samples of venous effluent were collected. The samples were analyzed to determine the fractional hydrolysis of the injected BPAP. The BPAP hydrolysis on passage through the lungs exhibited the saturable behavior and the relative insensitivity to changing flow rate previously described. Since we have described previously that BPAP behaves as if it exists in two forms, one of which is virtually unhydrolyzable on a single pass through the lungs, a model was formulated to include the influence of the unhydrolyzable form, as well as the saturable hydrolysis of the hydrolyzable form, on the fractional hydrolysis of the injected BPAP. This model provides a new method for estimating the kinetic parameters of BPAP hydrolysis by pulmonary endothelial ACE, and it explains the observation that the fractional BPAP hydrolysis does not vary with flow rate and transit time to the extent predicted by previous models. PMID- 2557774 TI - Application of the avidin-biotin-peroxidase complex (ABC-peroxidase) technique to the identification of human papillomavirus antigens on semithin sections by electron microscopy. AB - A post-embedding immunohistochemical method using avidin-biotin peroxidase complex (ABC-peroxidase) is described for localizing specific antigens for papillomavirus on semithin sections of human warts fixed and embedded in epoxy resins for conventional electron microscopy. The use of very specific and sensitive antibodies and the etching procedure with a saturated solution of sodium ethoxide appears to give good results. Results indicate that this method applied routinely on semithin sections, before the ultrastructural examination, may be suitable as a screening method for positive cells, which will be further examined by electron microscopy. PMID- 2557773 TI - Paraventricular nucleus modulation of glycemia and insulinemia in freely moving lean rats. AB - The effect of norepinephrine (NE) injection (40 nmol) into the paraventricular nucleus (PVN) on plasma insulin and glucose levels was studied in freely moving lean Zucker rats bearing chronic right jugular catheters for blood sampling and unilateral intracerebral cannulas placed just above the PVN. Already 2.5 min after NE injection, plasma glucose levels rose significantly, reaching a peak at 10 min poststimulus, whereas the insulin output was strongly inhibited. This NE induced hyperglycemia was independent of the corticosterone levels. A ganglionic blockade performed by intravenous chlorisondamine (1 mg/kg body wt) reduced by 80% the 4.5-min NE-induced incremental glucose areas. NE-induced hyperglycemia was reduced to a large extent when the PVN alpha-adrenergic receptors were blocked with phentolamine and to a lesser extent when the beta-adrenergic receptors were blocked with propranolol. NE-induced inhibition of insulin output was not affected by these adrenergic blockers. It is concluded that, when administered locally into the PVN, NE can activate the sympathetic outflow expressed by a neurally mediated hyperglycemia through central alpha- and beta adrenoreceptor and an inhibition of insulin output through other types of receptors and/or mechanisms. PMID- 2557775 TI - Concentration of vitamin B6 and activities of enzymes of B6 metabolism in the blood of alcoholic and nonalcoholic men. AB - The purpose of this study was to compare concentrations of vitamin B6 compounds and the activities of enzymes that synthesize or catabolize pyridoxal 5' phosphate in the plasma and erythrocytes of nonalcoholic and alcoholic subjects. Blood was obtained from male nonalcoholics and chronic alcoholics with minimal liver damage and normal hematology. Plasma, erythrocyte, and urinary B6 compounds were analyzed by high performance liquid chromatography, and pyridoxal phosphate was also measured enzymatically. Erythrocyte pyridoxine kinase and pyridoxine phosphate oxidase and erythrocyte and plasma pyridoxine phosphate phosphatases were assayed. Plasma pyridoxal phosphate concentration was significantly lower in the alcoholics (31.3 +/- 3.6 nmol/liter) than in the nonalcoholics (58.7 +/- 7.5 nmol/liter). The concentrations of the other B6 compounds in plasma, erythrocytes, and urine were not different in the two groups. Plasma alkaline pyridoxine phosphate phosphatase activity was significantly higher in the alcoholics (4.05 +/- 0.36 nmol/(h.mg] than in the nonalcoholics (3.01 +/- 0.18 nmol/(h.mg]. The activities of erythrocyte kinase, oxidase, and phosphatases were not significantly different in the two groups. The relationship of plasma pyridoxal phosphate concentration to its metabolites and the activities of the enzymes involved in its metabolism was determined. Plasma pyridoxine phosphate phosphatase activity assayed at pH 9.0 or 7.4 correlated negatively with plasma pyridoxal phosphate concentration. The low pyridoxal phosphate concentration observed in the plasma of the alcoholic subjects may in part be related to increased plasma phosphatase activity. PMID- 2557776 TI - Effects of ethanol on human platelets stimulated with platelet-activating factor, a biologically active ether phospholipid. AB - The interaction between ethanol and 1-0-alkyl-2-acetyl-sn-glycerol-3 phosphocholine (platelet activating factor, PAF) was addressed using platelets obtained from normal nonalcoholic volunteers. Ethanol at concentrations of 20 to 100 mM inhibited PAF activation of human platelets. Ethanol inhibited prominently the second or arachidonic acid metabolite dependent wave of platelet aggregation, which occurs with human platelets in citrated plasma. It also inhibited serotonin release and thromboxane A2 formation associated with this secondary phase of aggregation. Ethanol did not readily inhibit the primary wave of PAF-induced aggregation. The incorporation of PAF into platelets or metabolism of PAF was not influenced by up to 100 mM ethanol. Since ethanol inhibited only the secondary response, a direct interaction between PAF, ethanol, and a platelet PAF receptor is unlikely. The effect of ethanol on PAF-induced platelet aggregation shows a selectivity similar to that demonstrated by other investigators for epinephrine and adenosine diphosphate. PMID- 2557777 TI - A dot assay for the erythropoietin receptor using human recombinant 125I erythropoietin. AB - A dot assay was developed for the detection of membrane receptor(s) for erythropoietin (Ep). A relatively homogeneous population of cells bearing the receptor for Ep was generated in the spleen of mice made anemic with phenylhydrazine and crude membrane extracts were prepared from spleen cell suspensions. Aliquots of the membrane extracts were applied to microdishes of nitrocellulose in a volume of 4 microliters. After free reactive sites were blocked, the microdishes were incubated for 2 h at 37 degrees C with 125I-labeled human recombinant Ep (125I-rEp), and nitrocellulose bound radioactivity was determined thereafter. Reproducible curves were obtained, and a significant correlation between bound radioactivity and the amount of membrane proteins applied to the nitrocellulose dishes was found. Specific binding was saturable, reaching a plateau at 2.5 nM. Binding parameters of nitrocellulose-immobilized receptor were not significantly different from the values calculated using intact cells. No appreciable binding of 125I-rEp to control membranes at low Ep-receptor content was observed. Among a panel of growth factors, only unlabeled rEp was able to compete for the binding of 125I-rEp to nitrocellulose-immobilized membrane proteins in a dose-dependent fashion. The technique described herein may be of use in the study of the Ep receptor and as an assay for its purification. Moreover, it may also be of general application in the study of receptor-ligand interactions. PMID- 2557778 TI - Quantitative determination of SH groups in low- and high-molecular-weight compounds by an electron spin resonance method. AB - To quantitatively determine SH groups in high- and low-molecular-weight compounds, a disulfide biradical (RS-SR), where R is imidazoline residue, has been used. The biradical is shown to participate in a thiol-disulfide exchange reaction with compounds containing SH groups. In this case the ESR spectra of the biradical RS-SR and the resulting monoradical R-SH are different. The reaction of the biradical with cysteine, glutathione, and human serum albumin has been studied using the ESR method and the rate constants kf of this reaction have been calculated. Studies of the pH dependence of kf indicate that the thiol-disulfide exchange occurs by reaction with mercaptidione. Protein human serum albumin and hemoglobin have been modified by RS-SR. It has been shown that the treatment of modified proteins with reduced glutathione leads to removal of the radical from the protein; such modifications are thus reversible. The method proposed has been used to quantitatively determine the SH groups of cysteine and glutathione in mouse and rat blood. The method is shown to coincide within experimental error with the determination of glutathione and cysteine by titration with p chloromercuribenzoate or reaction with Ellman's reagent. This method allows detection of 10(-6)-10(-7) M SH compounds even in colored and highly absorbing samples. The kinetics of the SH group modification can also be determined, leading to deduction about accessibility of the SH group in protein. PMID- 2557779 TI - Magnetic separation of pinocytic vesicles of defined age from Entamoeba histolytica. AB - We describe a rapid and simple method to isolate pinocytic vesicles of defined age (residing time within the cell) from Entamoeba histolytica. Amoebas are allowed to pinocytize for greater than 5 min a suspension of superparamagnetic iron oxide particles, washed, and resuspended for predetermined periods (up to 150 min) in iron oxide-free medium. Subsequently, the cells are homogenized and iron oxide-containing vesicles are separated magnetically. Recovery of vesicles (estimated with fluorescein isothiocyanate-dextran as a quantitative marker for pinocytosis) was 20-40%. Contamination with "older" vesicles or with plasma membrane (estimated with fluorescein isothiocyanate-dextran and with fluorescein isothiocyanate-conjugated, succinylated concanavalin A, respectively) was negligible. Using this method we obtained evidence that in E. histolytica, contrary to the situation in animal cells, pinocytic vesicles within 150 min after invagination neither shrunk nor fused with each other to any significant extent. The method should be generally applicable to protozoa for the isolation of pinocytic vesicles and digestive vacuoles. PMID- 2557780 TI - [Incidence of local complications after spinal anesthesia in patients treated with low molecular weight heparin]. PMID- 2557781 TI - [Inhalation of smoke causes hydrocyanic acid intoxication]. PMID- 2557782 TI - [Anesthetic properties of a new inhibitor of glutaminergic transmission in the central nervous system, riluzole]. PMID- 2557783 TI - Asthmatic patients have neutrophils that exhibit diminished responsiveness to adenosine. AB - Activation of neutrophils (PMN) within the airways results in the secretion of a number of products such as reduced oxygen metabolites that could contribute to the inflammatory response associated with asthma. However, mediators of allergy, such as histamine, prostaglandin E2 (PGE2), isoproterenol, and adenosine, may serve to mitigate this inflammation through feedback inhibition of neutrophil function. To test the hypothesis that PMN activation and feedback inhibition mechanisms may be abnormal in asthmatics, we compared both superoxide production and adenosine-induced suppression of superoxide production in 12 matched pairs of asthmatics and control subjects. PMN obtained from asthmatic patients generated significantly more superoxide in response to f-met-leu-phe (fMLP) than controls (2.94 +/- 55 nmol/5 x 10(5) PMN/5 min versus 1.38 +/- 0.35 at 2 x 10(-8) M fMLP and 3.81 +/- 0.68 nmol versus 2.04 +/- 0.45 nmol at 10(-7) M; p less than 0.01 for both). In contrast, the respiratory burst generated by two receptor independent stimuli, the calcium ionophore A23187 and phorbol myristate acetate, was equivalent between control and asthmatic subjects. At 10(-6) M, 2 chloroadenosine induced a 19.5 +/- 5.1% inhibition of fMLP-stimulated superoxide production in PMN from patients with asthma as compared to 55.6 +/- 24.6% inhibition in PMN from control subjects (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557785 TI - Morphologic characterization of alveolar macrophages from subjects with occupational exposure to inorganic particles. AB - Alveolar macrophages recovered by bronchoalveolar lavage from 43 nonsmoking or greater than 5-yr ex-smoking subjects with occupational exposure to inorganic particles (asbestos, n 1/2 19; silica, n 1/2 10; coal, n 1/2 14) were evaluated by light microscopy and transmission and scanning electron microscopy to determine the morphologic changes resulting in these cells from chronic inorganic particulate inhalation. Alveolar macrophages from dust-exposed subjects, including those who had been free of exposure to particles for more than 1 yr, contained particles of higher proportion than did those of normal unexposed subjects. Most of these particles were located within phagolysosomes. The frequency of multinucleated alveolar macrophages was significantly higher in the dust-exposed groups. Ultrastructural studies showed alterations of the morphologic aspects of the surfaces of alveolar macrophages from the dust-exposed subjects, including increased numbers of rufflings, filopodia, pinocytotic vesicles, subplasmalemmal linear densities, and increased frequency of macrophage macrophage and macrophage-lymphocyte interactions. Furthermore, the numbers of lysosomes were significantly increased in alveolar macrophages from the dust exposed subjects. Together, these morphologic changes are consistent with the sequelae of phagocytosis, and they emphasize both the role of alveolar macrophages in eliminating inorganic particles from the alveolar spaces and the consequences this role has in alveolar macrophage activation. PMID- 2557784 TI - Viral pneumonia attenuates adenylate cyclase but not beta-adrenergic receptors in murine lung. AB - Respiratory infections provoke increased airway reactivity in both asthmatic and otherwise healthy subjects in part through impaired beta-adrenergic relaxation of bronchial and tracheal muscle. The precise mechanism remains obscure, but some studies report a decrease in the number of beta-adrenergic receptors. The present study was designed to assess the effect of viral respiratory infection with influenza A on lung beta-adrenergic receptors and adenylate cyclase activation in mice under two separate protocols. First, to determine whether changes are due to a local or systemic effect, we compared mice with influenza infections limited to the upper respiratory tract to mice with infection of the total respiratory tract. Four days after upper respiratory tract infection there were no changes in either isoproterenol- or NaF-stimulated adenylate cyclase activity. In contrast, there was an 82% decrease in isoproterenol- and a 25% decrease in NaF-stimulated adenylate cyclase activity on the fourth day after total respiratory tract infection. There were no changes in beta-adrenergic receptors or receptor coupling to adenylate cyclase with either type of infection. Our second protocol compared acutely infected mice to postrecovery mice. Twelve days after infection the virus was no longer present in the lungs, and adenylate cyclase activity was restored to normal. These data suggest that viral respiratory infection may impair airway function through attenuation of receptor and postreceptor activation of adenylate cyclase activity. PMID- 2557787 TI - Effect of the selective leukotriene B4 antagonist U-75302 on antigen-induced bronchopulmonary eosinophilia in sensitized guinea pigs. AB - The selective leukotriene B4 (LTB4) antagonist, U-75302, 6-(6-(3-hydroxy-1E,5Z undecadien-1-yl)-2-pyridinyl)-1,5-hexa nediol) was examined for its ability to inhibit the "late-phase" bronchopulmonary eosinophilia that occurs 6 to 24 h after inhalation of specific antigen in sensitized guinea pigs. Groups of 6 male guinea pigs, sensitized with ovalbumin, were pretreated with U-75302, 1.0, 10.0, or 30.0 mg/kg, or vehicle 1 h before and 7 h after antigen inhalation. Twenty four hours after antigen provocation, the lungs were lavaged for the enumeration of inflammatory cell populations. Doses of U-75302 (1.0, 10.0 and 30.0 mg/kg) administered orally produced 12.2%, (p greater than 0.05), 43.2% (p less than 0.05), and 61.1% (p less than 0.05) inhibition, respectively, of the antigen induced influx of eosinophils into the bronchial lumen. Neutrophil populations were not significantly affected by treatment with U-75302. In a separate study, we compared the histopathological changes that occurred following antigen challenge in U-75302-treated or vehicle-treated guinea pigs. Vehicle-treated, sensitized animals exhibited marked changes in the airway at 8 min, 6 h, and 24 h after antigen challenge. U-75302 treatment produced a significant reduction in eosinophil adherence to peribronchial/peribronchiolar capillaries followed by a dramatic and specific reduction of peribronchial eosinophil infiltration (81% reduction at 6 h and 79% reduction at 24 h). Neutrophil migration appeared unaffected. These data implicate LTB4 as a mediator of antigen-induced bronchopulmonary eosinophilia in the guinea pig. PMID- 2557786 TI - Methyl prednisolone acetate modulation of infection and subsequent pulmonary pathology in hamsters exposed to parainfluenza-1 virus (Sendai). AB - Parainfluenza-1 virus (PI-1) has a wide host range; the disease process observed varies with the age, previous exposure, and species. This study was performed to determine possible effects of the corticosteroid methyl prednisolone acetate (MPA) on PI-1 infection in hamsters. Hamsters serologically negative for PI-1 were exposed to virus alone or were exposed to virus the day after pretreatment with a single subcutaneous injection of MPA. Serum antibodies to PI-1 were present in virus-only exposed hamsters by Day 8 and increased up to Day 20. PI-1 was recovered from lungs of virus-only exposed hamsters on Day 2 to 8. Virus antigen was detected by immunocytochemistry on Days 2 to 10 in lungs of virus only exposed hamsters. Virus-associated lesions in these hamsters began as acute bronchiolar epithelium degeneration and necrosis on Day 4 and were foci of fibrosis by Days 12 to 20. Hamsters exposed to virus after MPA treatment developed no antibodies to virus, had no virus detectable by plaque assays or immunocytochemistry, and had no pulmonary lesions. Hamsters treated with MPA had decreased total lymphocyte counts up to Day 20 after treatment. Treatment of hamsters with MPA one day prior to PI-1 virus exposure is associated with no detectable evidence of viral infection. Humoral and cellular immunity mediated by MPA-sensitive lymphocytes may mediate some of the manifestations of PI-1 pulmonary disease. PMID- 2557788 TI - Dibutyryl-cAMP blocks endotoxin-induced lung injury in rats. AB - We investigated the effect of dibutyryl-cAMP pretreatment on endotoxin-induced hemodynamic changes and lung vascular injury in rats. In catheter-implanted, unanesthetized rats, intraperitoneal injection of Salmonella enteritidis endotoxin (2 mg/kg) decreased cardiac output and systemic blood pressure while increasing total pulmonary vascular resistance. Db-cAMP (1 mg given intraperitoneally every 30 min), although not significantly affecting cardiac output and systemic blood pressure, blocked the increase in total pulmonary resistance caused by endotoxin. Ninety minutes after intraperitoneal endotoxin injection, perfused lungs from endotoxin-treated rats exhibited increased pulmonary vascular permeability, as assessed by increased extravascular accumulation of 125I-albumin and water. Db-cAMP treatment in vivo markedly attenuated the increases in lung albumin leak index and wet-to-dry weight ratio caused by endotoxin without affecting lung microvascular pressures. This protective action of db-cAMP is not due to its effect on prostaglandin or leukotriene synthesis since endotoxin-stimulated increases in lung tissue 6-keto prostaglandin F1 alpha, thromboxane B2 and leukotriene C4 were not inhibited. We conclude that db-cAMP blocks endotoxin-induced lung injury in the rat by a mechanism independent of eicosanoid products and speculate that agents that increase intracellular cAMP may be therapeutically useful in acute lung vascular injury. PMID- 2557789 TI - Neutrophil elastase augments acute edematous injury in isolated rat lungs perfused with neutrophil cytoplasts. AB - To assess the contribution of neutrophil elastase to neutrophil-mediated acute edematous lung injury, isolated rat lungs were perfused with human neutrophil cytoplasts that produce O2 metabolites normally but contain little of the neutrophil elastase usually found in neutrophils. We found that addition of neutrophil cytoplasts and phorbol myristate acetate (PMA) caused less edematous injury in isolated lungs than addition of neutrophils and PMA. However, addition of neutrophil elastase along with neutrophil cytoplasts and PMA increased the amounts of lung injury found in lungs perfused with neutrophil cytoplasts and PMA to levels that equaled the amount of lung injury found after addition of normal neutrophils and PMA. By comparison, addition of purified neutrophil elastase, PMA alone, or human neutrophils alone did not cause lung injury. The results indicate that neutrophil elastase is a necessary component for the maximal development of acute edematous injury in isolated lungs perfused with neutrophils and PMA. PMID- 2557790 TI - Cystic nephroma: an emerging entity. AB - Cystic nephroma is a rare lesion, but there has been an increased number of cases reported in the recent literature. Most cases have been discovered incidentally either on routine examination or during radiological investigations. There are two peaks of incidence, one in childhood and one in middle age and the condition is often wrongly presumed to be a malignant tumor. Cystic nephroma is a benign neoplasm and it can be confidently diagnosed using modern imaging techniques and tissue analysis. Classically, the treatment of cystic nephroma has been simple nephrectomy, although a few cases have been treated by enucleation. Simple nephrectomy is the treatment of choice in children as the diagnosis in the young is less exact and a cure is assured. In some adult patients it is reasonable to pursue a conservative policy with regular clinical and ultrasound examinations. PMID- 2557791 TI - Effect of acute smoke inhalation on angiotensin converting enzyme, plasminogen activator, and angiotensin-II in the dog. AB - Smoke inhalation injuries in humans are associated with many uncontrolled variables which impact on the lung and make the cause of the pulmonary response difficult to assess. In this report, an established model of smoke inhalation injury in the dog was used to study the early responses of tissue and serum angiotensin-converting enzyme (ACE), tissue plasminogen activator (PLA), and plasma angiotensin II. Animals were exposed to smoke from burning sawdust and kerosene for five minutes. The hemodynamic and pulmonary mechanical responses were typical with a rise in pulmonary artery pressure, pulmonary vascular resistance, and venous admixture (shunt fraction) while dynamic compliance fell. Within five minutes of smoke exposure, lung ACE declined without any change in serum ACE. Lung PLA dropped one hour after injury. Plasma angiotensin II increased within 30 minutes without evidence for systemic hypertension. These early enzymatic changes substantiate the presence of pulmonary endothelial damage known to occur in this form of chemical injury. These changes may condition the lung's physiologic response to the injury and to additional stresses which are multiple when smoke inhalation occurs in conjunction with a cutaneous burn. PMID- 2557793 TI - Abnormal cough reflex in canine acrylamide neuropathy. AB - Coughing in response to irritation of the airways is a fundamental protective reflex that is dependent on rapidly adapting bronchopulmonary receptors and their vagal afferent fibers; reflex airway constriction, which is effected by vagomotor efferent fibers, usually accompanies coughing. Although dysfunction of vagally mediated cardiovascular and gastrointestinal reflexes is a well-documented complication of autonomic neuropathy, to date there have been no studies of the effect of peripheral autonomic failure on the cough reflex. In the study reported here, we examined the effect of acrylamide-induced neuropathy, a distal axonopathy, on the ventilatory and tracheomotor components of the cough reflex in conscious dogs. There was a reduction in the cough reflex in response to mechanical irritation of the large airways in the preclinical phase of the neuropathy, and the cough reflex was virtually abolished when the dogs had moderate neuropathy. Following withdrawal of the neurotoxin, there was a substantial recovery of the cough reflex in surviving animals. It is possible that the cough reflex may be reduced in patients with vagal neuropathy and that this might compromise protection of the airway. PMID- 2557792 TI - Abnormalities of the electron transport chain in idiopathic Parkinson's disease. AB - Idiopathic Parkinson's disease may have a low-level familial association but does not follow mendelian patterns of inheritance. Since inheritance of some components of the electron transport chain is nonmendelian and since inhibition of the electron transport chain with the toxin 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine models Parkinson's disease in humans and animals, we evaluated catalytic activities of the electron transport chain in platelet mitochondria purified from patients with idiopathic Parkinson's disease. All 10 patients studied had significant reductions of complex I (NADH:ubiquinone oxidoreductase) activity. Succinate:cytochrome c oxidoreductase activity was less strikingly reduced. We hypothesize that the complex I abnormality may have an etiological role in the pathogenesis of Parkinson's disease and that this defect may be derived via the mitochondrial genome. PMID- 2557794 TI - PET imaging of human gliomas with ligands for the peripheral benzodiazepine binding site. AB - Human gliomas were imaged in vivo using ligands for the peripheral-type benzodiazepine binding site (or omega 3 binding site) and positron emission tomography (PET). Although gliomas have a high density of the peripheral-type benzodiazepine binding site, PET scans with a selective ligand for this site, [11C] Ro5-4864, failed to demonstrate higher radioactivity levels in human gliomas than in brain. In vitro studies of surgically removed specimens of human glioma demonstrated little binding of Ro5-4864 but high levels of binding of another selective ligand, PK 11195. Scans with [11C]PK 11195 demonstrated increased radioactivity in glioma compared to brain in 8 of 10 patients. Radioactivity in tumor and the ratios of radioactivity in tumor to that in remote gray and in white matter correlated significantly with the specific activity of [11C]PK 11195, suggesting that accumulation represents saturable high-affinity binding. We conclude that the PK 11195 manifests greater binding than Ro5-4864 to the peripheral-type benzodiazepine binding site on human gliomas and that human gliomas can be successfully imaged using [11C]PK 11195 and PET. PMID- 2557795 TI - Subdivisional involvement of nigrostriatal loop in idiopathic Parkinson's disease and striatonigral degeneration. AB - A topographical immunocytochemical analysis was performed on the substantia nigra from patients with idiopathic Parkinson's disease and striatonigral degeneration. Antibodies to tyrosine hydroxylase, a marker for nigrostriatal dopaminergic neurons, and to calcineurin, a marker for striatonigral projection fibers, were used in this study. There was a marked depletion of dopaminergic neurons in the substantia nigra of parkinsonian patients compared with control subjects, the reduction being greater in the lateral portion than in the medial portion (p less than 0.001). Calcineurin immunoreactivity was densely distributed throughout the substantia nigra of patients with Parkinson's disease and control subjects. The numbers of dopaminergic neurons and of calcineurin-immunoreactive fibers were markedly reduced in the lateral portion of the substantia nigra in all patients with striatonigral degeneration. Our results suggest that many symptoms of these two diseases may be due to disruption of the functions of the putamen and the lateral portion of the substantia nigra, which have dense reciprocal connections as part of the dopamine-related nigrostriatal loop. PMID- 2557797 TI - Gm haplotypes in inflammatory demyelinating polyneuropathies. AB - The possible association of Gm haplotypes with inflammatory neuropathies has been studied in 59 patients with Guillain-Barre syndrome and 55 patients with chronic inflammatory demyelinating polyradiculoneuropathy. The frequency of Gm haplotype 1,2,17;21 (or z,a,x;g) was significantly raised in the patients with Guillain Barre syndrome. These findings provide further evidence for an association of chromosome 14 genes with inflammatory neuropathies. PMID- 2557796 TI - The widespread alteration of neurites in Alzheimer's disease may be unrelated to amyloid deposition. AB - The structural changes of Alzheimer's disease (AD) include a widespread alteration of neuronal cell processes in addition to senile plaques and neurofibrillary tangles. Since the antigenic characteristics of these abnormal neurites are similar to those of the abnormal neurites associated with the senile plaques, the question has been raised as to whether the widespread neuritic alteration is secondary to the deposition of amyloid. To answer this question, we examined brains from 2 subjects with a longer-lasting form of subacute sclerosing panencephalitis (SSPE) characterized by the presence of numerous neurofibrillary tangles but no senile plaques, 3 subjects with AD, and 2 age-matched controls. Light and electron immunocytochemical analyses revealed that abnormal neurites are present diffusely in SSPE cerebral cortex in the absence of amyloid deposits. These abnormal neurites were qualitatively identical to the widespread abnormal neurites of AD. The abnormal neurites, in contrast to the neurites of control brains, immunoreacted with antibodies to tau and ubiquitin. These distinctive antigenic features were due to the presence in these abnormal neurites of straight filaments, 14 to 16 nm in diameter, mixed with a few paired helical filaments. The spatial distribution of the widespread neuritic alteration correlated with that of neurofibrillary tangles in both conditions, but not with that of senile plaques in AD. The present findings demonstrate that a diffuse alteration of neurites similar to that present in AD takes place independently of the deposition of amyloid in SSPE, and they are consistent with the hypothesis that in AD, also, this alteration is not secondary to the deposition of amyloid.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557798 TI - Competitiveness of different polysaccharide utilization mutants of Bacteroides thetaiotaomicron in the intestinal tracts of germfree mice. AB - Bacteroides thetaiotaomicron, an obligate anaerobe found in high numbers in human colons, can utilize a variety of polysaccharides. To determine which type of polysaccharide contributes most to the nutrition of B. thetaiotaomicron in vivo, we isolated and characterized transposon-generated mutants deficient in the ability to use different polysaccharides. Some mutants were deficient in polysaccharide utilization because of the inability to utilize a component monosaccharide. These mutants included a mutant that was unable to utilize L fucose (a component of goblet cell mucin), a mutant that was unable to utilize D galactose (a component of raffinose, stachyose, arabinogalactan, and goblet cell mucin), and a mutant that was unable to utilize either glucuronic acid (a component of mucopolysaccharides) or galacturonic acid (a component of polygalacturonic acid or pectin). Other mutants were unable to use the polysaccharide but could use the component sugars. These included four mutants that were unable to utilize starch and one mutant that was unable to utilize polygalacturonic acid. The mutants were tested for the ability to compete with the wild type for colonization of the intestinal tracts of germfree mice. The only mutants against which the wild type competed successfully in the intestinal tracts of germfree mice were a galactose-negative mutant and a uronic acid negative mutant. These mutations differed from the others tested in that they affected utilization of more than one type of polysaccharide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557799 TI - Serial 99mTc dimercaptosuccinic acid (DMSA) scans after urinary infections presenting before the age of 5 years. AB - Forty five children presenting with a first proven urinary tract infection under the age of 5 years were studied by sequential 99mTc dimercaptosuccinic acid (DMSA) scans. Forty nine kidneys in 40 children had definite defects at presentation, and 39 (80%) of these defects were still present when the DMSA scan was repeated. Changes in the appearance of defects were independent of the presence or degree of reflux at presentation and of symptomatic recurrence of infection, though the combination of new infection and grade 3 reflux (reflux reaching the renal calices with distension) was associated with deterioration. No kidney with a relative DMSA uptake of less than 35% showed any improvement in its cortical defects. Only two kidneys that were initially without defects, in a single patient who had bilateral grade 3 reflux and breakthrough infections, developed defects on subsequent scans. The outcome after urinary tract infection is dependent on the effect of the first infection on the kidney. Occasionally children with grade 3 reflux develop damage during subsequent infections. More widespread use of DMSA scans should improve our understanding of the factors that determine the development of renal damage. PMID- 2557800 TI - Formulation of malaria treatment policy for children in Cote d'Ivoire as chloroquine resistant Plasmodium falciparum spreads into west Africa. AB - To develop a malaria treatment policy for children with Plasmodium falciparum, an in vivo and in vitro chloroquine (CQ) sensitivity study was conducted in Cote d'Ivoire in September 1986. The efficacy of a single dose of CQ (10 mg base kg-1, C10) was tested with assessment of subjects on Days 2 and 7 after treatment; 108 (99%) of 109 children were aparasitaemic on Day 7. Of 33 isolates of P. falciparum tested in vitro, two (6%) were resistant to CQ. Although C10 appeared effective clinically and parasitologically in Cote d'Ivoire, a treatment dose of 25 mg of CQ base kg-1 (C25), over three days, was recommended as first-line therapy for malaria. This was because in vivo CQ-resistance will soon spread into other West African countries including Cote d'Ivoire, the C25 dose still retains clinical effectiveness in most partially-immune persons living in areas with low level chloroquine resistance, and alternate drugs are more expensive. PMID- 2557801 TI - Schistosoma mansoni in Sierra Leone: an invader extending its range? PMID- 2557802 TI - Detection of Entamoeba histolytica trophozoites in liver pus by the indirect fluorescent antibody test for the aetiological diagnosis of amoebic liver abscess. AB - The indirect fluorescent antibody test was used to detect trophozoites of Entamoeba histolytica in liver pus aspirated from patients with amoebic liver abscess. The test can be carried out in no more than two hours. Trophozoites with fluorescence were observed in 17 of the 18 patients with amoebic liver abscesses who were studied. Cells with fluorescence were not found in any negative control specimens from patients with bacterial liver abscess, primary liver cancer, cirrhosis or tuberculous peritonitis. These results indicate that this sensitive, specific and rapid test is very useful in aetiological diagnosis of amoebic liver abscess. PMID- 2557803 TI - Evidence for selection of virulent sub-populations of Entamoeba histolytica by cholesterol. AB - Quantitatively much higher Concanavalin A (Con. A) agglutinability, haemolytic potency, and activities of acid hydrolases, namely phosphatase (EC 3.1.3.2), ribonuclease (EC 2.7.7.16), deoxyribonuclease (EC 3.1.4.5) and proteinase--were observed in a virulent strain of Entamoeba histolytica (IP-106), as compared to attenuated and avirulent strains (200-NIH) and DKB respectively. In addition, significant differences in these parameters were observed among clonal cultures derived from the latter two cultures by cultivation of single amoebic cells picked out by micromanipulation. Repeated sub-culturing of parent cultures of both these strains in cholesterol-enriched medium resulted in marked enhancement of all the above activities, but no such change occurred in the derived clonal cultures following similar cholesterol treatment. The implication of these findings in relation to enhancement of the virulence of E. histolytica by cholesterol is discussed. PMID- 2557804 TI - The effect of pentostam and cimetidine on the development of leishmaniasis (Leishmania mexicana amazonensis) and concomitant malaria (Plasmodium yoelii). AB - BALB/c mice were infected with Leishmania mexicana amazonensis and/or Plasmodium yoelii in order to determine the impact of multiple parasitic infection on the efficacy of chemotherapeutic agents. Uninfected, P. yoelii-infected, L.m. amazonensis-infected, and L.m. amazonensis and P. yoelii-infected mice were inoculated with cimetidine (80 mg kg-1 day-1) or pentostam (200 mg kg-1 day-1) once a day for an initial 20-day period, and once a week thereafter. Leishmania mexicana amazonensis lesion development and P. yoelii parasitaemia were the criteria used to assay disease severity. Mice infected with both P. yoelii and L.m. amazonensis developed more severe disease than did animals infected with either parasite alone. Cimetidine and pentostam each slowed the development of L.m. amazonensis in animals infected with only that parasite and in animals infected with both P. yoelli and L.m. amazonensis. However, mice treated with pentostam developed more severe P. yoelii infections than did control animals, whereas cimetidine significantly reduced P. yoelii parasitaemia in all instances. PMID- 2557805 TI - [Hypoglycemic action of different doses of nopal (Opuntia streptacantha Lemaire) in patients with type II diabetes mellitus]. AB - To assess the relationship between the doses of O. streptacantha Lem. and its acute hypoglycemic action in diabetics, eight patients with type II diabetes mellitus were studied. Four test were performed to each patient with the intake of: (a) 400 ml of water, (b) 100 g (c) 300 g and (d) 500 g of broiled stems of O. streptacantha Lem. Serum glucose was measured at 0, 60, 120 and 180 minutes. Maximal decrease of serum glucose was noticed at 180 minutes, with a mean of 2.3, 10, 30.1 and 46.7 mg/dl less than basal value with 0, 100, 300 and 500 g respectively (P = NS, less than 0.05, less than 0.001 and less than 0.001 respectively). A significant direct correlation (r = 0.690, P less than 0.001) was noticed between the doses and the hypoglycemic effect. PMID- 2557806 TI - Depression of the vestibulo-ocular and optokinetic responses by intrafloccular microinjection of GABA-A and GABA-B agonists in the rabbit. AB - The functional implication of the cerebellar flocculus in regulation of the VOR and OKR gain has mostly been studied by lesion experiments, and the hypotheses derived from these experiments are not always in line with one another. In the present study, a reversible method was used to inhibit floccular Purkinje cells. The GABA-A agonist muscimol or the GABA-B agonist baclofen were bilaterally injected into the flocculus of rabbits, and the effects of these injections on the gain of the VOR and OKR were studied. Both drugs induced a reduction by at least 50% of the gain of the VOR in light and darkness, and of the OKR. Although GABA-A and GABA-B receptors are known to have different cerebellar localizations, muscimol and baclofen injections resulted in quantitatively similar effects. It is suggested that these GABA-agonists cause either direct or indirect inhibition of floccular Purkinje cells, thus reducing modulation of the firing rate of these neurons by afferent mossy and climbing fibers. Because the flocular Purkinje cells act out of phase with the vestibular neurons which drive the oculomotor neurons, a reduced output of floccular Purkinje cells would result in a reduction of the VOR and OKR gain. These experiments provide strong evidence that the cerebellar flocculus has a positive influence on the basic VOR and OKR gain. PMID- 2557808 TI - On 1,4-benzodiazepines and 1,5-benzodiazocines, XII: Solution-state conformation of KC-2846 and its interaction with hypothetical benzodiazepine receptor groups. PMID- 2557807 TI - Injections of a beta-adrenergic antagonist in pontine reticular structures modify the gain of vestibulospinal reflexes in decerebrate cats. AB - 1. The norepinephrine (NE)-containing locus coeruleus (LC) neurons control posture as well as the gain of the vestibulospinal reflexes either through direct coeruleospinal (CS) projections or by inhibiting the dorsal pontine reticular formation (pRF) and the related medullary inhibitory reticulospinal (RS) system. The question whether these inhibitory influences on the pRF are mediated through beta-adrenoceptors was investigated by injecting in precollicular decerebrate cats small doses of the non-selective beta-adrenergic antagonist propranolol in different pontine tegmental structures. 2. Injection of propranolol (usually 0.25 microliters at the concentration of 4.5 micrograms/microliters of saline) in dorsal pontine structures, which decreased the tonic contraction of limb extensors ipsilateral to the side of the injection, greatly increased the amplitude of the multiunit EMG responses of the ipsilateral triceps brachii to roll tilt of the animal at 0.15 Hz, +/- 10 degrees. Correspondingly, the response gain of the forelimb extensor to labyrinth stimulation increased. Moreover, a slight decrease in phase lead of the responses was observed. These responses were always characterized by an increased EMG activity during ipsilateral tilt and a decreased activity during contralateral tilt, as shown in the control records (alpha-responses). The same injection also produced in some instances an increase of the extensor tonus of the contralateral limbs, associated with an increased EMG activity of the contralateral triceps brachii; on the other hand, the amplitude of modulation and thus the response gain of this muscle to the same parameters of labyrinth stimulation decreased, while the response pattern reversed (beta-responses), thus being opposite to that displayed by the triceps brachii ipsilateral to the side of the injection. 3. The changes in posture and reflexes described above appeared 10-20 min after unilateral injection of propranolol in the pRF, reached in highest values in about 60-100 min and persisted for more that three hours before returning to the control level. These effects were not due to irritative phenomena following injection of the fluid, since neither changes in posture nor in the response gain of the triceps brachii to labyrinth stimulation were observed after injection of an equal volume of saline in the pRF of that side. Moreover, the magnitude of the effects increased to some extent in relation to the dose of the beta-adrenergic blocker. 4. Histological controls indicated that the structure responsible for these postural and reflex changes was located in the dorsal pontine tegmental region immediately ventral to the LC and included the peri-LC alpha and the surrounding dorsal pRF.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557809 TI - Rapid identification and typing of herpes simplex virus by a new enzyme immunoassay with peroxidase-labeled complement C1q. AB - A new enzyme-linked immunoassay system (TC-ELISA) was developed for the rapid, specific detection and typing of herpes simplex virus (HSV) using anti-HSV immune sera and enzyme-labeled complement component C1q (P*-C1q). The method is based on viral antigen produced in HSV-infected cells being detected by simultaneous addition of immune serum and P*-C1q (ELISA-CF). With this assay, it was found that HSV was detected with anti-HSV immune serum and P*-C1q and that HSV type 1 and HSV type 2 could be differentiated with anti-HSV-1 and anti-HSV-2 immune sera and P*-C1q. The TC-ELISA test was HSV specific and not reactive with measles virus, cytomegalovirus, mumps virus or varicella-zoster virus. The whole assay procedure can be completed within 3 hours, as long as HSV-inoculated cell cultures with more than 10% cytopathic effect are used. These results suggest that TC-ELISA is useful in the diagnosis of viral infections. PMID- 2557810 TI - Pattern of anti-cytomegalovirus IgM antibodies determined by immunoblotting. A study of kidney graft recipients developing a primary or recurrent CMV infection. AB - In order to improve the knowledge of the humoral immune response to CMV infection, we developed an immunoblotting technique which allowed a better analysis of the changes in the pattern of anti CMV-polypeptides IgM. We examined 234 sera belonging to 27 renal allograft recipients developing a primary or recurrent CMV infection and 12 non infected recipients. Thus we found that 11 main anti CMV-polypeptides IgM antibodies were present in over 25% of the infected patients. They reacted with proteins whose molecular weights ranged from 32K to 205K. We showed that anti-p 45-47 IgM antibodies were present in 100% of CMV infected recipients and never in the non-infected population. They appeared very early in the course of the infection (5.43 weeks post-graft for primary infection and 5.00 weeks for recurrent ones) and, therefore, constitute a good marker of active infection. Two other CMV-specific IgM antibodies (anti-p 60-64 and anti-p 100) were found exclusively in the course of primary infections. Anti p 60-64 IgM was observed at a high frequency (57.1%) and with a mean delay of 6.57 weeks post-graft. Therefore, the anti-p 60-64 IgM detection could be helpful for the diagnosis of primary infection. In almost 100% of both primary and recurrent infections, we observed anti-p 140 and anti-p 38 IgM antibodies. Only about 50% of non-infected patients had low levels of anti-p 140 and anti-p 38 IgM. The follow-up of recurrent infections showed that the anti CMV-polypeptides IgM antibodies appeared earlier than in primary infection. When we compared anti p 45-47 IgM detection by immunoblotting and anti-CMV IgM detected by ELISA we observed that immunoblotting permitted the diagnosis 2.5 weeks earlier for primary infection, and 1 week earlier for recurrent infection, than ELISA. In addition, the detection of anti-p 45-47 IgM antibodies also occurred earlier than virus excretion. PMID- 2557811 TI - Detection of pseudorabies virus DNA sequences by the polymerase chain reaction. AB - Genomic sequences of pseudorabies virus, a porcine herpesvirus, were amplified by the polymerase chain reaction from cells of infected cultures, nasal cells and organs from acutely diseased as well as from organs of latently infected pigs. PMID- 2557812 TI - [Adrenergic coronary vasoconstriction during myocardial hypoperfusion]. AB - The intramural branches of the coronary arteries of dogs were studied using venilite casts. The intramyocardial vessels have a general pattern with straight branches that cross from the epicardium toward the endocardium. Many branches were observed throughout the extension of these vessels, mainly in the subendocardium, where there is a rich interconnecting plexus. Some branches divides immediately in the subepicardium. These anatomic findings concur with the observations that the subendocardium is more susceptible to ischemia, as it is dependent on an extensive vascularization. These anatomic findings indicate that during constant underperfusion, the alpha adrenergically mediated vasoconstriction of the subepicardial vessels have an unexpected beneficial effect because of decreased transmural steal. PMID- 2557813 TI - [Cardiovascular effects of omega-3 fatty acids: its significance in preventing arteriosclerosis and coronary disease]. PMID- 2557814 TI - [The role of angiotensin-converting enzyme inhibitor (captopril) on the mechanism of hypoxic pulmonary vasoconstriction. Experimental study in dogs]. AB - In order to evaluate the action of an angiotensin converting enzyme inhibitor (Captopril) on the pulmonary hypoxic vasoconstriction, twenty one mongrel dogs were studied in two groups: group I with hypoxia, group II with normoxia. The dogs were anesthetized, intubated, and had their femoral vein and artery cannulated for blood-gas sampling and pressure records. They were mechanically ventilated with hypoxic gas mixtures (12.3% O2-87.7% N2)--group I and room air group II, at random. In both groups we measured, before and after administration of captopril 3 mg/kg intravenously, gas exchange and hemodynamic variables, as well as plasmatic levels of renin and angiotensin converting enzymes (ACE). Our results showed that the group I dogs decreased the systemic and pulmonary vascular resistances with small changes in pulmonary arterial pressures and no significant variations of pulmonary systemic resistances ratio. There were no significant variations of the same variables in the group II dogs. The gas exchange has not changed in either group of animals. In the group I dogs Captopril provoked systemic and pulmonary vasodilatation, with no gasometric and ventilation/perfusion ratio changes. In our experimental model we could not conclude that Captopril inhibited the hypoxic pulmonary vasoconstriction and/or that the angiotensin II had some action on the hypoxic pulmonary vasoconstriction mechanism, but there are some evidences favoring that hypothesis. PMID- 2557815 TI - Effects of bicarbonate-containing versus lactate-containing peritoneal dialysis solutions on superoxide production by human neutrophils. AB - Human neutrophils were isolated from healthy volunteers and exposed to either bicarbonate-containing (pH 7.4) or lactate-containing (pH 5.2) peritoneal dialysis solution in vitro. Superoxide production by neutrophils was measured by a method based on the superoxide dismutase-inhibitable reduction of ferricytochrome c. Bicarbonate-containing peritoneal dialysis solution was found to be superior to the lactate-containing one in facilitating the production of superoxide anion by human neutrophils. PMID- 2557816 TI - Magnetic resonance in obstructive jaundice. AB - Twelve cases of obstructive jaundice in whom ultrasound failed to demonstrate the site and/or the cause of obstruction of the biliary tract were examined with magnetic resonance imaging (MRI), correctly diagnosing the site and cause of obstruction in 10 of 12 surgically proven cases. In one case of cholangiocarcinoma, the site of obstruction was well shown on MR but a definite cause could not be ascertained. In another patient who developed intermittent jaundice following surgery for choledochal cyst, MR demonstrated a solitary stone in the common hepatic duct. Surgical confirmation could not be achieved as the patient was lost to follow up. There were 6 cases of choledocholithiasis, 3 cases of gall bladder carcinoma and one case each of pancreatic adenocarcinoma and cholangiocarcinoma. It is believed that MRI will provide obstructive jaundice and will be able to minimize the use of percutaneous transhepatic cholangiography (PTC) and endoscopic retrograde cholangiopancreatography (ERCP) in view of its ability to perform multiplanar imaging in multiple sequences. PMID- 2557817 TI - Primary malignant fibrous histiocytoma of the stomach: (a case report). AB - A case of primary malignant fibrous histiocytoma (MFH) of the stomach is reported. The patient was a 51-year-old man who had a laparotomy for a malignant gastric tumour diagnosed by radiological studies and endoscopic examination. An inoperable gastric malignancy was found at operation. Histopathological studies of tissue biopsied at surgery showed characteristic light microscopic findings and strong positivity for alpha-1-antitrypsin by immunoperoxidase technique indicating the diagnosis of malignant fibrous histiocytoma. The absence of any other soft tissue or retroperitoneal tumour strongly indicated the stomach is the primary site. PMID- 2557818 TI - Papillomavirus infection of the female genital tract before and after treatment: a cytological, colposcopic and histological study. AB - Papillomavirus infection (HPV) of the female genital tract is now recognized as a major risk factor for the development of neoplasia. With the combined investigative modalities of cytology, colposcopy and histopathology, it is clear that precancerous changes and HPV are closely associated. The differentiation of innocent HPV infection from progressive premalignant disease is not clear-cut in all situations. Despite various treatment modalities, close follow-up of these patients by cytology, colposcopy and histology reveals the presence of persistent genital tract infection by HPV. PMID- 2557819 TI - Phospholipase C-mediated hydrolysis of phosphatidylcholine is activated by muscarinic agonists. AB - The phospholipase C-catalysed breakdown of inositol-containing phospholipids is an important source of diacylglycerol in cells stimulated by several agonists. However, recent experimental evidence suggests that major phospholipids such as phosphatidylcholine may also be substrates of the phosphodiesteratic hydrolysis activated by hormones, growth factors and oncogene products. We show here that stimulation of muscarinic agonists activates the release of phosphocholine, which, along with diacylglycerol, is a metabolic product of phospholipase C mediated hydrolysis of phosphatidylcholine. Fluoroaluminates mimic this muscarinic effect, strongly suggesting that carbachol-activated release of phosphocholine may be mediated by a guanine-nucleotide-binding protein. Evidence for this was obtained from experiments using permeabilized cells in which non hydrolysable analogues of GTP activated phosphocholine release synergistically with carbachol. PMID- 2557820 TI - Bradykinin and thrombin effects on polyphosphoinositide hydrolysis and prostacyclin production in endothelial cells. AB - Prostacyclin (PGI2) production by thrombin- and bradykinin-stimulated bovine aortic endothelial cells (BAEC) and human umbilical vein endothelial cells (HUVEC) was related to the receptor-linked activation of inositide hydrolysis. Bradykinin caused a rapid and transient 3-fold increase in the formation of inositol polyphosphates in BAEC. The increase in InsP3 reflected changes mainly in the Ins(1,4,5)P3 isomer. Thrombin was less effective than bradykinin in increasing InsP3 levels and appeared to only minimally stimulate the production of PGI2 in BAEC. In HUVEC, thrombin caused a 5-fold elevation of Ins(1,4,5)P3, closely related to a rise in PGI2 production. However, bradykinin did not affect inositol phosphates and PGI2 production in HUVEC. Other inositol phosphates were also assessed to obtain information on putative metabolism of Ins(1,4,5)P3. The present study supports the notion that formation of Ins(1,4,5)P3 is linked to an increase in PGI2 production in endothelial cells and furthermore provides evidence for a large degree of heterogeneity in the responses of BAEC and HUVEC to thrombin and bradykinin. PMID- 2557821 TI - Secondary structural features of the bacteriophage Mu-encoded A and B transposition proteins. AB - The role of the bacteriophage Mu-encoded A and B proteins is to direct the transposition of Mu DNA. These are the first active DNA transposition proteins to have been purified and their mechanism of action at the biochemical level is under intensive study. Structural studies on these proteins, however, have lagged behind their biochemical characterization. We report here near- and far-u.v. c.d. spectra for these proteins and their secondary structural features derived from these data. The Mu A protein appears to be composed of primarily beta-sheet (40%) with 24% alpha-helix, 9% beta-turn and 27% random coil. In contrast, the Mu B protein contains 55% alpha-helix with only 13% beta-sheet and 3+ beta-turn and 29% random coil. The near-u.v. c.d. spectrum of the A protein was not unusual; however, the profile of the B protein suggested either buried or restricted chromophores within the protein or short-range interactions between aromatic residues. PMID- 2557822 TI - Fragments of human fibroblast collagenase. Purification and characterization. AB - On purification, human fibroblast collagenase breaks down into two major forms (Mr22,000 and Mr 27,000) and one minor form (Mr 25,000). The most likely mechanism is autolysis, although the presence of contaminating enzymes cannot be excluded. From N-terminal sequencing studies, the 22,000-Mr fragment contains the active site; differential binding to concanavalin A shows the 25,000-Mr fragment is a glycosylated form of the 22,000-Mr fragment. These low-Mr forms can be separated by Zn2+-chelate chromatography. An activity profile of this column, combined with data from substrate gels, indicates no activity against collagen in the 22,000-Mr and 25,000-Mr forms, but rather, activity casein and gelatin. The 27,000-Mr form has no activity. The 22,000/25,000-Mr form can act as an activator for collagenase in a similar way to that reported for stromelysin. The activity of the 22,000/25,000-Mr form is not inhibited by the tissue inhibitor of metalloproteinases (TIMP). The 27,000-Mr C-terminal part of the collagenase molecule therefore appears to be important in maintaining the substrate specificity of the enzyme, and also plays a role in the binding of TIMP. PMID- 2557823 TI - Properties of a microtubule-associated cofactor-independent protein kinase from pig brain. AB - A protein kinase activity was identified in pig brain that co-purified with microtubules through repeated cycles of temperature-dependent assembly and disassembly. The microtubule-associated protein kinase (MTAK) phosphorylated histone H1; this activity was not stimulated by cyclic nucleotides. Ca2+ plus calmodulin, phospholipids or polyamines. MTAK did not phosphorylate synthetic peptides which are substrates for cyclic AMP-dependent protein kinase, cyclic GMP dependent protein kinase. Ca2+/calmodulin-dependent protein kinase II, protein kinase C or casein kinase II. MTAK activity was inhibited by trifluoperazine [IC50 (median inhibitory concn.) = 600 microM] in a Ca2+-independent fashion. Ca2+ alone was inhibitory [IC50 = 4 mM). MTAK was not inhibited by heparin, a potent inhibitor of casein kinase II, nor a synthetic peptide inhibitor of cyclic AMP-dependent protein kinase. MTAK demonstrated a broad pH maximum (7.5-8.5) and an apparent Km for ATP of 45 microM. Mg2+ was required for enzyme activity and could not be replaced by Mn2+. MTAK phosphorylated serine and threonine residues on histone H1. MTAK is a unique cofactor-independent protein kinase that binds to microtubule structures. PMID- 2557825 TI - Ubiquitin-protein conjugates accumulate in the lysosomal system of fibroblasts treated with cysteine proteinase inhibitors. AB - Mouse fibroblasts (3T3-L1 cells) accumulate detergent- and salt-insoluble aggregates of proteins conjugated to ubiquitin when incubated in the presence of inhibitors of lysosomal cysteine cathepsins, including E-64. These ubiquitin protein conjugates co-fractionate with lysosomes on density gradients and are found in multivesicular dense bodies which by electron microscopy appear to be engaged in microautophagy. Both E-64 and ammonium chloride increase the intracellular concentration of free ubiquitin, but only E-64 leads to the formation of insoluble lysosomal ubiquitin-protein conjugates. The results are discussed in relation to the possible intracellular roles of ubiquitin conjugation. PMID- 2557824 TI - Characterization of hepatic lactogen receptor. Subcellular distribution and characterization of N-linked carbohydrate chains. AB - The types of carbohydrate chains present in a rat liver lactogenic hormone binding receptor species with an Mr of 82,000, and in its hormone-binding subunits with Mr values of 40,000 and 35,000, were characterized using carbohydrate-chain-cleaving enzymes and affinity cross-linking. The subcellular distribution of lactogenic hormone-binding species was studied in organelle enriched fractions. The monomeric Mr-40,000 and Mr-35,000 species contain N linked tri- or tetra-antennary complex and high-mannose chains respectively. The Mr-82,000 species exists in two forms, where the Mr-40,000 and Mr-35,000 subunits are each combined with unglycosylated and, with the technique used, unlabelled subunit(s). Studies with organelle-enriched fractions revealed that the Mr-35,000 species was found in an endoplasmic reticulum-enriched fraction. The Mr-40,000 species was the predominant monomeric binding species in Golgi/endosome- and plasma membrane-enriched fractions. It is suggested that the Mr-35,000 species is a precursor to the Mr-40,000 species. In lysosome/endosome- or lysosome-enriched fractions, a broad distribution in Mr (35,000-40,000) was characteristic of the hormone-binding species. The Mr-82,000 species was only found in a Golgi/endosome enriched fraction. Labelling of endosome lactogen receptor by injection of 125I labelled ovine prolactin in vivo and cross-linking yielded only the Mr-40,000 species. Thus, the Mr-40,000 and Mr-35,000 lactogenic hormone-binding species each appear to be combined with the unglycosylated receptor subunit(s) in the Golgi complex to form Mr-82,000 heterodimeric complexes. PMID- 2557826 TI - Cloning and expression in Escherichia coli of a rat hepatoma cell cDNA coding for 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase. AB - In liver, the 470-residue bifunctional enzyme 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase (PFK-2/FBPase-2) catalyses the synthesis and degradation of fructose 2,6-bisphosphate, a potent stimulator of glycolysis. In rat hepatoma (HTC) cells, this enzyme has kinetic, antigenic, and regulatory properties, such as insensitivity to cyclic AMP-dependent protein kinase and lack of associated FBPase-2 activity, that differ from those in liver. To compare the sequence of the HTC enzyme with that of the liver enzyme, we have cloned the corresponding fully-coding cDNA from HTC cells. This cDNA predicts a protein of 448 residues in which the first 32 residues of liver PFK-2/FBPase-2 including the cyclic AMP target sequence have been replaced by a unique N-terminal decapeptide. The rest of the protein is identical with the liver enzyme. An N-terminally truncated recombinant peptide of 380 residues containing the PFK-2 and FBPase-2 domains was expressed in Escherichia coli as a beta-galactosidase fusion protein. It was recognized by anti-PFK-2 antibodies but its enzymic activities were barely detectable. In contrast, a cDNA fully-coding for the HTC enzyme could be expressed in E. coli as a beta-galactosidase-free peptide that exhibited both PFK 2 and FBPase-2 activities. This peptide had those PFK-2 kinetic properties of the HTC enzyme that differ from the liver enzyme. These data, together with immunoblot experiments, suggest that the lack of associated FBPase-2 activity in HTC cells results from a post-translational modification of the enzyme rather than from the difference in amino acid sequence. As well as this peculiar type of PFK-2/FBPase-2 mRNA, HTC cells also contained low concentrations of the liver type mRNA. Unlike in liver, neither mRNA was induced by dexamethasone in these cells. PMID- 2557827 TI - Mapping the benzodiazepine photoaffinity-labelling site with sequence-specific gamma-aminobutyric acidA-receptor antibodies. AB - The gamma-aminobutyric acidA (GABAA) receptor purified from adult bovine cerebral cortex was photoaffinity-labelled with the agonist benzodiazepine [3H]flunitrazepam and the radioactivity shown to be coincident with a band with Mr 53,000 that was recognized by three anti-(GABAA receptor alpha 1 subunit sequence)-specific antibodies. Complete and limited CNBr cleavage of the purified photoaffinity-labelled receptor was carried out. The products of this reaction were analysed for radioactivity, for immunoreactivity with anti-[alpha 1-(1-15) peptide], anti-[alpha 1-(324-341)-peptide] and anti-[alpha 1-(413-429)-peptide] polyclonal antibodies and for carbohydrate by biotinylated concanavalin A lectin overlay. Complete CNBr cleavage gave a radioactive peptide with Mr 10,000-12,000 that was not recognized by the above-mentioned specific antisera. By using the deduced amino acid sequence of the alpha 1 subunit [Schofield, Darlison, Fujita, Burt, Stephenson, Rodriguez, Rhee, Ramachandran, Reale, Glencorse, Seeburg & Barnard (1987) Nature (London) 328, 221-227], it is proposed that the site of the benzodiazepine-agonist photoaffinity-labelling reaction does not lie within the amino acid sequences alpha 1 1-58 and alpha 1 149-429. PMID- 2557828 TI - Cyclosporin A inhibits phorbol ester-induced activation of superoxide production in resident mouse peritoneal macrophages. AB - Peritoneal resident macrophages from mice are sensitive to inhibition by cyclosporin A (CsA) of phorbol 12-myristate 13-acetate (PMA)-stimulated oxidative burst. Inhibition was assessed in terms of superoxide anion (O2.-) and H2O2 production. Key findings were as follows. (a) CsA inhibited in a dose-dependent manner the production of O2.- when cells were stimulated with PMA. CsA did not alter the respiratory burst induced by other stimuli (zymosan, concanavalin A and fMet-Leu-Phe). It was verified that CsA itself had no scavenger effect. (b) A concomitant decrease in H2O2 liberation following CsA exposure was found. This inhibition was observed both in the initial rate of synthesis and in the accumulation after 15 min of incubation. (c) NADPH oxidase activity in the crude supernatant was unaffected by the previous incubation of macrophages with CsA. CsA does not inhibit glucose transport measured as 14CO2 production. (d) The production of O2.- was strongly dependent on the glucose concentration. Sodium oleate also stimulated O2.- production in resident macrophages. These data might be correlated with the inhibitory effect of CsA upon other functions of macrophages. PMID- 2557830 TI - Site-directed mutagenesis of the Klebsiella pneumoniae nitrogenase. Effects of modifying conserved cysteine residues in the alpha- and beta-subunits. AB - The five conserved cysteine residues present in the alpha-subunit and the three conserved cysteine residues present in the beta-subunit of nitrogenase component 1 were individually changed to alanine. Mutations in the alpha-subunit at positions 63, 89, 155 and 275 and in the beta-subunit at positions 69, 94 and 152 all resulted in a loss of diazotrophic growth and component 1 activity and loss of the normal e.p.r. signal of the component 1 protein. Component 2 activity was retained. Replacement of cysteine-184 in the alpha-subunit with alanine greatly diminished, but did not eliminate, diazotrophic growth and component 1 activity. Substitution of serine for cysteine at position 152 in the beta-subunit, in contrast with the substitution of alanine at this position, resulted in the formation of active component 1. Replacement of the non-conserved cysteine-112 in the beta-subunit with alanine did not greatly perturb diazotrophic growth or the activity of component 1. Extracts prepared from a mutant, with cysteine-275 of the alpha-subunit replaced by alanine, complemented extracts of a mutant unable to synthesize the iron-molybdenum cofactor of nitrogenase, indicating that the alanine-275 substitution increases the availability of cofactor. Furthermore extracts of this mutant exhibited an e.p.r. signal similar to that of extracted iron-molybdenum cofactor. These data suggest a role for cysteine-275 as a ligand to the cofactor. PMID- 2557829 TI - Superoxide generation is inhibited by phospholipase A2 inhibitors. Role for phospholipase A2 in the activation of the NADPH oxidase. AB - The stimulation of O2.- generation by phorbol 12-myristate 13-acetate (PMA) in human neutrophil-derived cytoplasts was inhibited by a variety of phospholipase A2 inhibitors in a concentration-dependent manner. Inhibition was found to be independent of the order of addition of the inhibitor and PMA. The most potent inhibitor, RO 31-4639, inhibited O2.- generation with an IC50 value (concentration causing 50% inhibition) of 1.5 microM. The addition of either arachidonic acid or SDS, in the presence of the inhibitors, was able to restore O2.- generation. The results suggest that arachidonic acid, released by phospholipase A2, is necessary for both the activation and the maintenance of O2. generation by the NADPH oxidase. PMID- 2557831 TI - Electrochemical and spectroscopic characterization of the 7Fe form of ferredoxin III from Desulfovibrio africanus. AB - Desulfovibrio africanus ferredoxin III is a monomeric protein (Mr 6585) containing seven cysteine residues and 7-8 iron atoms and 6-8 atoms of acid labile sulphur. It is shown that reversible unmediated electrochemistry of the two iron-sulphur clusters can be obtained by using a pyrolytic-graphite-'edge' carbon electrode in the presence of an appropriate aminoglycoside, neomycin or tobramycin, as promoter. Cyclic voltammetry reveals two well-defined reversible waves with E0' = -140 +/- 10 mV and -410 +/- 5 mV (standard hydrogen electrode) at 2 degrees C. Bulk reduction confirms that each of these corresponds to a one electron process. Low-temperature e.p.r. and magnetic-c.d. spectroscopy identify the higher-potential redox couple with a cluster of core [3Fe-4S]1+.0 and the lower with a [4Fe-4S]2+.1+ centre. The low-temperature magnetic-c.d. spectra and magnetization properties of the three-iron cluster show that it is essentially identical with that in Desulfovibrio gigas ferredoxin II. We assign cysteine-11, 17 and -51 as ligands of the [3Fe-4S] core and cysteine-21, -41, -44 and -47 to the [4Fe-4S] centre. PMID- 2557832 TI - Electrochemical and spectroscopic characterization of the conversion of the 7Fe into the 8Fe form of ferredoxin III from Desulfovibrio africanus. Identification of a [4Fe-4S] cluster with one non-cysteine ligand. AB - Desulfovibrio africanus ferredoxin III is a protein (Mr 6585) containing one [3Fe 4S]1+,0 and one [4Fe-4S]2+,1+ core cluster when aerobically isolated. The amino acid sequence contains only seven cysteine residues, the minimum required to ligand these two clusters. Cyclic voltammery by means of direct electrochemistry at a pyrolytic-graphite-'edge' electrode promoted by neomycin shows that, when reduced, the [3Fe-4S]0 centre reacts rapidly with Fe(II) ion to form a [4Fe-4S]2+ cluster. The latter, which can be reduced at a redox potential similar to that of the other [4Fe-4S] cluster, must include non-thiolate ligation. We propose that the carboxylate side chain of aspartic acid-14 is the most likely candidate, since this amino acid occupies the position of a cysteine residue in the sequence typical of an 8Fe ferredoxin. The magnetic properties at liquid-He temperature of this novel cluster, studied by low-temperature magnetic-c.d. and e.p.r. spectroscopy, are diamagnetic in the oxidized state and S = 3/2 in the one electron-reduced state. This cluster provides a plausible model for the ligation states of the [4Fe-4S]1+ core in the S = 3/2 cluster of the iron protein of nitrogenase and in Bacillus subtilis glutamine:phosphoribosyl pyrophosphate amidotransferase. PMID- 2557833 TI - Membrane-entrapped microperoxidase as a 'solid-state' promoter in the electrochemistry of soluble metalloproteins. AB - Immobilization of biological systems in solid matrices is presently of great interest, in view of the many potential advantages associated with both the higher stability of the immobilized macromolecules and the potential utilization for biotechnology. In the present paper the electrochemical behaviour of the undecapeptide from cytochrome c (called microperoxidase) tightly entrapped in cellulose triacetate membrane is reported; its utilization as 'solid-state' promoter in the electrochemistry of soluble metalloproteins is presented. The results obtained indicate that: (i) membrane-entrapped microperoxidase undergoes rapid reversible electron transfer at a glassy carbon electrode; (ii) the electrochemical process is diffusion-controlled; (iii) entrapped microperoxidase acts as 'solid-state' promoter in the electrochemistry of soluble cytochrome c and of azurin. PMID- 2557834 TI - Characterization of two proteolytically derived soluble polypeptides from the neurofilament triplet components NFM and NFH. AB - We have purified to homogeneity the regions derived by chymotryptic digestion of the ox neurofilament polypeptides NFH and NFM; the regions, called M1 and M2, are thought to form part of the projecting sidearms of mammalian neurofilaments [Chin, Eagles & Maggs (1983) Biochem. J. 215, 239-252]. They were isolated and purified under non-denaturing conditions and showed no tendency to interact with each other in solution. The Mr values obtained by sedimentation are approx. 61,000 for M1 and 42,000 for M2, considerably lower than the values obtained by SDS/polyacrylamide-gel electrophoresis. These Mr values were unchanged in the presence of 6 M-guanidine hydrochloride, suggesting that the regions exist as monomers in solution. Both M1 and M2 are highly phosphorylated, and there is only a slight change in the sedimentation value upon dephosphorylation. Dephosphorylation of M1 with alkaline phosphatase was more than 90% efficient but was never absolute. Dephosphorylation of M2 was complete. Both M1 and M2 bind Ca2+; in the case of M1, this binding is phosphorylation-dependent. M1 also binds cytochrome c, and dephosphorylation affects binding. In similar conditions, neurofilaments bind at least twice their own mass of cytochrome c, owing to their opposite net charges. No interactions were observed between native or dephosphorylated M1 and M2, and intact neurofilaments under a wide variety of conditions. These results are discussed in terms of the possible roles that neurofilament sidearms might play and throw doubt upon their supposed function of rigidly cross-linking neurofilaments together within the axoplasm of neurons. PMID- 2557835 TI - Induction of ketogenesis and fatty acid oxidation by glucagon and cyclic AMP in cultured hepatocytes from rabbit fetuses. Evidence for a decreased sensitivity of carnitine palmitoyltransferase I to malonyl-CoA inhibition after glucagon or cyclic AMP treatment. AB - The effects of pancreatic hormones and cyclic AMP on the induction of ketogenesis and long-chain fatty acid oxidation were studied in primary cultures of hepatocytes from fetal and newborn rabbits. Hepatocytes were cultivated during 4 days in the presence of glucagon (10(-6) M), forskolin (2 x 10(-5) M), dibutyryl cyclic AMP (10(-4) M), 8-bromo cyclic AMP (10(-4) M) or insulin (10(-7) M). Ketogenesis and fatty acid metabolism were measured using [1-14C]oleate (0.5 mM). In hepatocytes from fetuses at term, the rate of ketogenesis remained very low during the 4 days of culture. In hepatocytes from 24-h-old newborn, the rate of ketogenesis was high during the first 48 h of culture and then rapidly decreased to reach a low value similar to that measured in cultured hepatocytes from term fetuses. A 48 h exposure to glucagon, forskolin or cyclic AMP derivatives is necessary to induce ketone body production in cultured fetal hepatocytes at a rate similar to that found in cultured hepatocytes from newborn rabbits. In fetal liver cells, the induction of ketogenesis by glucagon or cyclic AMP results from changes in the partitioning of long-chain fatty acid from esterification towards oxidation. Indeed, glucagon, forskolin and cyclic AMP enhance oleate oxidation (basal, 12.7 +/- 1.6; glucagon, 50.0 +/- 5.5; forskolin, 70.6 +/- 5.4; cyclic AMP, 77.5 +/- 3.4% of oleate metabolized) at the expense of oleate esterification. In cultured fetal hepatocytes, the rate of fatty acid oxidation in the presence of cyclic AMP is similar to the rate of oleate oxidation present at the time of plating (85.1 +/- 2.6% of oleate metabolized) in newborn rabbit hepatocytes. In hepatocytes from term fetuses, the presence of insulin antagonizes in a dose-dependent fashion the glucagon-induced oleate oxidation. Neither glucagon nor cyclic AMP affect the activity of carnitine palmitoyltransferase I (CPT I). The malonyl-CoA concentration inducing 50% inhibition of CPT I (IC50) is 14-fold higher in mitochondria isolated from cultured newborn hepatocytes (0.95 microM) compared with fetal hepatocytes (0.07 microM), indicating that the sensitivity of CPT I decreases markedly in the first 24 h after birth. The addition of glucagon or cyclic AMP into cultured fetal hepatocytes decreased by 80% and 90% respectively the sensitivity of CPT I to malonyl-CoA inhibition. In the presence of cyclic AMP, the sensitivity of CPT I to malonyl-CoA inhibition in cultured fetal hepatocytes is very similar to that measured in cultured hepatocytes from 24-h-old newborns. PMID- 2557837 TI - Purification and properties of uroporphyrinogen III synthase (co-synthase) from an overproducing recombinant strain of Escherichia coli K-12. AB - The Escherichia coli hemD gene, encoding the enzyme uroporphyrinogen III synthase (co-synthase), was cloned into multi-copy plasmids in E. coli cells that were used to generate strains producing up to 1000 times the concentration of the synthase in the wild-type. The enzyme was purified to homogeneity from these strains in milligram amounts. The enzyme is a monomer of Mr 28,000 with an isoelectric point of 5.2 and a pH optimum of 7.8. The specific activity of the purified synthase is 1500 units/mg and the Km for the substrate, pre uroporphyrinogen, is 5 microM. The N-terminal sequence of the enzyme is Ser-Ile Leu-Val-Thr-Arg-Pro-Ser-Pro-Ala-Gly-, in agreement with the gene-derived protein sequence. The enzyme contains four 5,5'-dithiobis-(2-nitrobenzoic acid) titratable groups, one reacting rapidly with the reagent and three further groups having lower reactivity. The enzyme is heat-sensitive, and during heat inactivation all four thiol groups become equally available for reaction. PMID- 2557838 TI - Modification of myo-inositol monophosphatase by the arginine-specific reagent phenylglyoxal. AB - myo-Inositol monophosphatase is inhibited by the arginine-specific reagent phenylglyoxal. The rate of inactivation is decreased in the presence of Pi, a competitive inhibitor of the enzyme. The effect of Pi is dependent on the presence of Mg2+, but is unaffected by Li+, an uncompetitive inhibitor. In the absence of Mg2+, the substrate, Ins(1)P, binds to the enzyme but is not converted into products, and affords only a small degree of protection against inactivation by phenylglyoxal. Li+ had no further effect under these conditions, but in the presence of Mg2+ caused a marked potentiation of the protective effect of substrate alone. In the absence of substrate, Li+ had no effect on activation by phenylglyoxal. Incorporation of 14C-labelled phenylglyoxal showed that inactivation was associated with modification of a single arginine residue per monomer in the dimeric enzyme. These findings support a mechanism in which Li+ inhibits monophosphatase by trapping a phosphorylated enzyme intermediate and preventing its hydrolysis. PMID- 2557839 TI - Decreased activity and impaired hormonal control of protein phosphatases in rat livers with a deficiency of phosphorylase kinase. AB - 1. Livers from gsd/gsd rats, which do not express phosphorylase kinase activity, also contain much less particulate type-1 protein phosphatases. In comparison with normal Wistar rats, the glycogen/microsomal fraction contained 75% less glycogen-synthase phosphatase and 60% less phosphorylase phosphatase activity. This was largely due to a lower amount of the type-1 catalytic subunit in the particulate fraction. In the cytosol, the synthase phosphatase activity was also 50% lower, but the phosphorylase phosphatase activity was equal. 2. Both Wistar rats and gsd/gsd rats responded to an intravenous injection of insulin plus glucose with an acute increase (by 30-40%) in the phosphorylase phosphatase activity in the liver cytosol. In contrast, administration of glucagon or vasopressin provoked a rapid fall (by about 25%) in the cytosolic phosphorylase phosphatase activity in Wistar rats, but no change occurred in gsd/gsd rats. 3. Phosphorylase kinase was partially purified from liver and subsequently activated. Addition of a physiological amount of the activated enzyme to a liver cytosol from Wistar rats decreased the V of the phosphorylase phosphatase reaction by half, whereas the non-activated kinase had no effect. The kinase preparations did not change the activity of glycogen-synthase phosphatase, which does not respond to glucagon or vasopressin. Furthermore, the phosphorylase phosphatase activity was not affected by addition of physiological concentrations of homogeneous phosphorylase kinase from skeletal muscle (activated or non activated). 4. It appears therefore that phosphorylase kinase plays an essential role in the transduction of the effect of glucagon and vasopressin to phosphorylase phosphatase. However, this inhibitory effect either is specific for the hepatic phosphorylase kinase, or is mediated by an unidentified protein that is a specific substrate of phosphorylase kinase. PMID- 2557836 TI - Role of guanine nucleotide regulatory proteins in insulin stimulation of glucose transport in rat adipocytes. Influence of bacterial toxins. AB - The potential role of guanine nucleotide regulatory proteins (G-proteins) in acute insulin regulation of glucose transport was investigated by using bacterial toxins which are known to modify these proteins. Cholera-toxin treatment of isolated rat adipocytes had no effect on either 2-deoxyglucose transport or insulin binding. Pertussis-toxin treatment resulted in an inhibition of both insulin binding and glucose transport. Insulin binding was decreased in pertussis toxin-treated cells by up to 40%, owing to a lowering of the affinity of the receptor for hormone, with no change in hormone internalization. The dose response curve for insulin stimulation of glucose transport was strongly shifted to the right by pertussis-toxin treatment [EC50 (half-maximally effective insulin concn.) = 0.31 +/- 0.04 ng/ml in control cells; 2.29 +/- 1.0 in treated cells), whereas cholera toxin had only a small effect (EC50 = 0.47 +/- 0.02 ng/ml). Correcting for the change in hormone binding, pertussis toxin was found to decrease the coupling efficiency of occupied receptors (50% of maximal insulin effect with 928 molecules bound/cell in control and 3418 in treated cells). Pertussis-toxin inhibition of insulin sensitivity was slow in onset, requiring 2 3 h for completion. Under conditions where pertussis-toxin inhibition of insulin sensitivity was maximal, a 41,000 Da protein similar to the alpha subunit of Gi (the inhibitory G-protein) was found to be fully ribosylated. These results are consistent with the concept that pertussis-toxin-sensitive G-protein(s) can modify the insulin-receptor/glucose-transport coupling system. PMID- 2557841 TI - The local anaesthetic benzyl alcohol attenuates the alpha 2-adrenoceptor-mediated inhibition of human platelet adenylate cyclase activity when stimulated by prostaglandin E1, but not that stimulated by forskolin. AB - Treatment of human platelets with concentrations of benzyl alcohol up to 50 mM augmented adenylate cyclase activity when it was assayed in the basal state and when stimulated by prostaglandin E1 (PGE1), isoprenaline or NaF. Benzyl alcohol antagonized the stimulatory effect exerted on the catalytic unit of adenylate cyclase by the diterpene forskolin. Benzyl alcohol did not modify the magnitude of the inhibitory response when the catalytic unit of adenylate cyclase was inhibited by using either low concentrations of guanosine 5'-[beta gamma imido]triphosphate, which acts selectively on the inhibitory guanine nucleotide regulatory protein Gi, or during alpha 2-adrenoceptor occupancy, by using adrenaline (+ propranolol). Some 34% of the potent inhibitory action of adrenaline on PGE1-stimulated adenylate cyclase was obliterated in a dose dependent fashion (concn. giving 50% inhibition = 12.5 mM) by benzyl alcohol, with the residual inhibitory action being apparently resistant to the action of benzyl alcohol at concentrations up to 50 mM. Treatment of membranes with benzyl alcohol did not lead to the release of either the alpha-subunit of Gi or G protein subunits. The alpha 2-adrenoceptor-mediated inhibition of adenylate cyclase was abolished when assays were performed in the presence of Mn2+ rather than Mg2+ and, under such conditions, dose-effect curves for the action of benzyl alcohol on PGE1-stimulated adenylate cyclase activity were similar whether or not adrenaline (+propranolol) was present. We suggest that (i) alpha 2-adrenoceptor- and Gi-mediated inhibition of PGE1-stimulated adenylate cyclase may have two components, one of which is sensitive to inhibition by benzyl alcohol, and (ii) the Gi-mediated inhibition of forskolin-stimulated adenylate cyclase exhibits predominantly the benzyl alcohol-insensitive component. PMID- 2557840 TI - Neutrophil degranulation inhibits potential hydroxyl-radical formation. Relative impact of myeloperoxidase and lactoferrin release on hydroxyl-radical production by iron-supplemented neutrophils assessed by spin-trapping techniques. AB - Hydroxyl radical (.OH) formation by neutrophils in vitro requires exogenous iron. Two recent studies [Britigan, Rosen, Thompson, Chai & Cohen (1986) J. Biol. Chem. 261, 17026-17032; Winterbourn (1987) J. Clin. Invest. 78, 545-550] both reported that neutrophil degranulation could potentially inhibit the formation of .OH, but differed in their conclusions as to the responsible factor, myeloperoxidase (MPO) or lactoferrin (LF). By using a previously developed spin-trapping system which allows specific on-line detection of superoxide anion (O2-) and .OH production, the impact of MPO and LF release on neutrophil .OH production was compared. When iron-diethylenetriaminepenta-acetic acid-supplemented neutrophils were stimulated with phorbol myristate acetate or opsonized zymosan, .OH formation occurred, but terminated prematurely in spite of continued O2- generation. Inhibition of MPO by azide increased the magnitude, but not the duration, of .OH formation. No azide effect was noted when MPO-deficient neutrophils were used. Anti-LF antibody increased both the magnitude and duration of .OH generation. Pretreatment of neutrophils with cytochalasin B to prevent phagosome formation did not alter the relative impact of azide or anti-LF on neutrophil .OH production. An effect of azide or anti-LF on spin-trapped-adduct stability was eliminated as a confounding factor. These data indicate that neutrophils possess two mechanisms for limiting .OH production. Implications for neutrophil-derived oxidant damage are discussed. PMID- 2557842 TI - Heparin binds to Leishmania donovani promastigotes and inhibits protein phosphorylation. AB - We show that promastigotes of Leishmania donovani, the causative agent of visceral leishmaniasis (kala-azar), possess heparin receptors on their surface. From a linear Scatchard plot of the binding data obtained using [3H]heparin and viable promastigotes, one derives a binding constant of 4.7 x 10(-7) M and an estimate of 860,000 receptors per parasite. The [3H]heparin bound to parasites could not be displaced by hyaluronic acid or by three other glycosaminoglycans (dermatan sulphate, chondroitin 4-sulphate and chondroitin 6-sulphate). It was demonstrated that exponential phase promastigotes growing in medium 199 supplemented with fetal bovine serum incorporate 35SO4 into a cell-associated macromolecule that has the properties of heparin proteoglycan. Heparin inhibits the activity of the cell-surface histone-protein kinase; incubation of viable promastigotes with [gamma-32P]ATP and MgCl2 (10 mM) in the absence and presence of heparin (0.01-0.5 mg/ml) for 10 min, followed by analysis by SDS/polyacrylamide-gel electrophoresis and autoradiography, revealed that the phosphorylation of 12 or 13 parasite proteins was inhibited by the glycosaminoglycan. These data suggest that heparin may play a role in the host parasite relationship. PMID- 2557843 TI - Phosphatidylinositol 4,5-bisphosphate hydrolysis in human sperm stimulated with follicular fluid or progesterone is dependent upon Ca2+ influx. AB - Hydrolysis of the phospholipid phosphatidylinositol 4,5-bisphosphate is thought to be intimately involved in agonist-induced changes in intracellular Ca2+ levels. Recently we have shown that human preovulatory follicular fluid, which induces exocytosis in human sperm, can stimulate a rapid, transient increase in sperm cytosolic [Ca2+] [Thomas & Meizel (1988) Gamete Res. 20, 397-411]. We report here that both a Sephadex G-75 column fraction, derived from follicular fluid, and progesterone (a component of both the G-75 fraction and whole follicular fluid) stimulate rapid hydrolysis of PtdIns(4,5)P2 and PtdIns4P in human sperm. We also report that progesterone stimulates a rapid influx of Ca2+ in human sperm. Human spermatozoa were labelled for 24 h with myo-[3H]inositol and then treated with either the G-75 fraction or progesterone. A 30-65% loss of label was detected in PtdIns(4,5)P2 and PtdIns4P within 15 s of stimulus addition; no changes were observed in PtdIns during 2 min of treatment. The loss of label from both lipids was accompanied by an increase in water-soluble inositol phosphates. Production of both InsP3 and InsP2 was seen within 10 s; however, InsP3 was rapidly removed and had reached control levels by 1 min. Similarly, formation of InsP2 reached a peak by 30 s and then began a decline accompanied by a corresponding increase in InsP. No increases in InsP4 were seen in sperm treated in this fashion. Stimulated hydrolysis of the phosphoinositides and release of inositol phosphates were both blocked by the Ca2+ antagonist La3+. Likewise, the progesterone-induced increase in intracellular Ca2+ was inhibited by La3+, and phosphoinositide hydrolysis stimulated by this hormone was dependent upon the presence of extracellular Ca2+. PMID- 2557844 TI - Functional heterogeneity of polyphosphoinositides in human erythrocytes. AB - After labelling of erythrocytes with [32P]P1 for 23 h, the specific radioactivities of the phosphomonoester groups of PtdIns4P and of PtdIns(4,5)P2 approached equilibrium values which were close to that of the gamma-phosphate of ATP (78-85%), showing that almost all of these phosphate groups were metabolically active. Phosphoinositidase C (PIC) activation, using Ca2+ and the ionophore A23187, of 32P-prelabelled erythrocytes was used to investigate a possible functional heterogeneity of the phosphoinositides. Hydrolysis of PtdIns(4,5)P2, measured from its radioactivity, decreased as function of the time of prelabelling up to a constant value equal to that measured from its content. In contrast, hydrolysis of PtdIns4P, determined both from radioactivity and from content, was always the same. These data suggest that newly labelled molecules of PtdIns(4,5)P2, initially accessible to PIC, then moved towards a PIC-resistant pool. This was further confirmed by measuring the fraction of labelled PtdIns(4,5)P2 molecules accessible to PIC after a prelabelling period of 5 min and different times of reincubation. Hydrolysis by PIC was also measured in erythrocytes in which the phosphoinositide content had been modified by activation (Mg2+-enriched cells) or inhibition (ATP-depleted cells) of the phosphoinositide kinases. The sizes of the PIC-resistant pools of polyphosphoinositides were not affected by these treatments, indicating that the kinases (and the phosphatases) act on the PIC-sensitive pools. This was also shown by the decrease in the production of Ins(1,4,5)P3 upon PIC activation in ATP-depleted erythrocytes. A model is presented in which the PIC-sensitive pools of polyphosphoinositides are those which are accessible to the kinases and the phosphatases and are rapidly turned over. PMID- 2557845 TI - Role of arginine residues in the stimulation of the smooth-muscle plasma-membrane Ca2+ pump by negatively charged phospholipids. AB - Negatively charged phospholipids strongly stimulate the purified plasma membrane Ca2+ pump of erythrocytes [Enyedi, Flura, Sarkadi, Gardos & Carafoli (1987) J. Biol. Chem. 262, 6425-6430] and of smooth muscle [Missiaen, Raeymaekers, Wuytack, Vrolix, De Smedt & Casteels, (1989) Biochem. J. 263, 687-694]. We have investigated the role of arginine residues in the interaction of these acidic phospholipids with the smooth-muscle Ca2+ transport ATPase. The arginine modifying reagent phenylglyoxal inhiibited the ATPase activity in a time dependent fashion by decreasing the Vmax. of the Ca2(+)-activation curve. Low concentrations of PtdIns, PtdIns4P, PtdIns(4,5) P2, phosphatidylserine and phosphatidic acid partially prevented this inactivation. This protective effect was however not apparent at higher concentrations of PtdIns4P, PtdIns(4,5) P2 and phosphatidic acid, which may be related to the previously observed inhibition of the enzyme at higher concentrations of these phospholipids. These findings indicate that the functionally important interaction of the acidic lipids with the protein occurs at least partially via arginine residue(s). PMID- 2557847 TI - Apparent inactivation of alpha 1-antiproteinase by sulphur-containing radicals derived from penicillamine. AB - alpha 1-Antiproteinase is the major inhibitor of proteolytic enzymes, such as elastase, in human plasma. Its elastase-inhibitory capacity can be inactivated by exposure to hydroxyl radicals (.OH) generated either by pulse radiolysis or by an Fe3+-EDTA/H2O2/ascorbic acid system. Inactivation of alpha 1-antiproteinase by radiolytically-generated .OH under anoxic conditions was decreased by adding a range of anti-inflammatory drugs to the reaction mixtures, including the thiol compound penicillamine. However, under conditions favouring formation of oxysulphur radicals, protection by thiols such as penicillamine was much decreased. It is proposed that sulphur-containing radicals resulting from attack of biologically-produced oxidants upon penicillamine in the presence of O2 can themselves inactivate alpha 1-antiproteinase, and that such radicals might contribute to the side-effects produced by penicillamine or gold thiol therapy in rheumatoid arthritis. PMID- 2557846 TI - Phospholipase D activation is functionally linked to superoxide generation in the human neutrophil. AB - Neutrophils stimulated with formylmethionyl-leucylphenylalanine (fMet-Leu-Phe) in the presence of butanol and ethanol formed phosphatidyl alcohols through a phospholipase D mechanism. The alcohols inhibited phosphatidic acid and diradylglycerol (DRG) formation, but did not block inositol 1, 4, 5-trisphosphate release. fMet-Leu-Phe-stimulated superoxide production was inhibited by alcohol concentrations which blocked DRG formation, whereas opsonized-zymosan-stimulated superoxide production was only partially decreased. These results suggest that phospholipase D activation is functionally linked to superoxide production in the human neutrophil. PMID- 2557849 TI - Induction by the calcium ionophore A23187 of a protective effect against cell injury in cultured gastric mucosal cells. AB - An intrinsic protective mechanism against cell injury seems to exist in cultured gastric mucosal cells. Cells, isolated from the stomachs of 10- to 12-day-old rats and subcultured, were examined for damage by the erythrosine B dye exclusion test. Pretreatment with 5 microM A23187 (a calcium ionophore) diminished the cell damage induced by acidified medium (pH 3.5) or 8 mM aspirin (pH 5.0). The effect of A23187 appeared 4 hr after its addition and was reversible. Protection by A23187 against cell injury diminished in the absence of extracellular Ca2+ and was dependent on Ca2+ concentration. An increase in intracellular Ca2+ may induce cell resistance against injury in cultured gastric mucosal cells. PMID- 2557848 TI - Influence of cardioprotective cyclooxygenase and lipoxygenase inhibitors on peroxidative injury to myocardial-membrane phospholipid. AB - Oxygenase-catalyzed and non-enzymatic polyunsaturated fatty acid peroxidations have potential pathogenic roles in ischemic-reperfusion damage to the myocardium. Certain oxygenase inhibitors protect heart muscle from irreversible ischemic injury, and some antiperoxidants can inhibit oxygenase enzymes. We investigated the antiperoxidative abilities of eight anti-ischemic, cardioprotective oxygenase inhibitors to prevent myocardial-membrane phospholipid peroxidation through superoxide-driven, iron-promoted reactions with xanthine oxidase as the source of superoxide. Flurbiprofen, ibuprofen, and REV-5901-5 did not affect peroxidation at concentrations up to 1000 microM. BW755C, AA-861, nafazatrom, dipyridamole, and propyl gallate did protect and cardiac lipids against oxidative injury in a concentration-dependent manner with respective and antiperoxidant IC50 values (concentrations at which peroxidation was inhibited by 50%) of 0.22, 1.25, 3.0, 3.6 and 50 microM. Catechin and phenidone, known oxygenase inhibitors not yet evaluated as anti-ischemic agents, were also found to be antiperoxidants at low micromolar concentrations. Four cyclooxygenase inhibitors ineffective against myocardial infarction (aspirin, indomethacin, naproxen, and sulfinpyrazone) evidenced no antiperoxidant properties at concentrations up to 500 microM. The oxygenase inhibitor-antiperoxidants identified could neither quench superoxide radical nor inhibit xanthine oxidase. However, they were able to interrupt the propagation of an on-going peroxidation reaction. Their antiperoxidant profiles resembled those of known antioxidants, such as alpha-tocopherol, which inhibit peroxidation by intercepting lipid free-radical intermediates. These data raise the possibility that at least some oxygenase inhibitors could exert cardioprotective effects by directly influencing the sensitivity of myocardial membrane phospholipid to peroxidative injury. Consequently, recognition of the antiperoxidant properties of these agents may aid dissection of their physiological and pharmacological actions. PMID- 2557850 TI - Method of extracting DNA from fine needle aspirates of human solid tumors for Southern blot analysis. AB - A rapid, simple, convenient method for extracting DNA from fine needle aspiration (FNA) samples of human solid tumors for Southern blot hybridization studies is described. After the preparation of an air-dried cytologic smear, the remaining sample in the needle was rinsed directly into a test tube for DNA extraction. The extraction procedure, in which manipulation of the sample is minimized, produced sufficient DNA for Southern blot analysis within 24 hours of the FNA biopsy in the ten consecutive cases studied. The DNA bound to the nylon membranes can be washed and reexamined with a variety of probes, allowing studies of lymphoid cell lineage, oncogene amplification or tumor progression. The assessment of cellularity on the cytologic specimen at the time of FNA provided a reliable guide to the need for further passes to obtain sufficient cells for DNA hybridization; the cytologic diagnosis could also be made on the smears. PMID- 2557851 TI - Methodologic aspects of DNA assessment by means of image cytometry in tumors of the salivary glands. A comparison between the results obtained using sections and cytospin preparations from the same paraffin-embedded specimens. AB - The image cytometric nuclear DNA assessments on paraffin-embedded tissue sections and on Cytospin preparations of disaggregated specimens from the same cases were compared in 98 salivary gland tumors, including 21 acinic cell carcinomas, 29 mucoepidermoid carcinomas, 21 adenocarcinomas and 27 adenoid cystic carcinomas. The histogram type (diploid, tetraploid or aneuploid) and the number of cells with DNA values greater than 2.5c (expressed in relative units) were considered as variables in the correlation. A high correlation between the results in different specimens was found in acinic cell carcinomas, mucoepidermoid carcinomas and adenocarcinomas; the histogram type and the number of cells with DNA values greater than 2.5c were essentially the same between specimen types in these three tumor entities. The cases of adenoid cystic carcinomas showed a considerably lower degree of correlation: in 8 of the 27 cases, the Cytospin preparations yielded diploid histograms, while the tissue sections yielded aneuploid histograms. The number of cells with DNA values greater than 2.5c was notably lower in the Cytospin preparations from adenoid cystic carcinoma; the reasons for this exceptional behavior of the cells of adenoid cystic carcinoma are discussed. These findings demonstrate that paraffin-embedded specimens of different tumor entities, even from the same organ, can be affected differently by disaggregation procedures. While retrospective studies on disaggregated paraffin-embedded specimens can yield reliable results, comparative assessments using both DNA analysis techniques, as in this study, should be performed before a large number of cases is evaluated. PMID- 2557852 TI - Cytologic, histologic, DNA ploidy and electron microscopic analyses of a case of vulvar Paget's disease. AB - The cells in a case of multifocal vulvar Paget's disease were studied by cytology, histology, ploidy analysis and electron microscopy. The Paget cells in smears were seen in an isolated or a sheetlike arrangement. The cells had peripherally located nuclei, prominent nucleoli and unevenly distributed melanin granules of various sizes in their cytoplasms. The histologic sections contained multifocal microscopic lesions that were larger than the macroscopic lesion. Paget cells in the histologic sections demonstrated positive cytoplasmic staining with the periodic acid-Schiff, Alcian blue, mucin, glandular cytokeratin, epithelial membrane antigen and carcinoembryonic antigen reactions. The nuclear DNA histogram of the Paget cells in the cytologic smears showed a polyploid pattern, with a sharp peak at 4c; the cells in sheets had a ploidy level between 2c and 4c while a few of the isolated cells had a ploidy level that extended past 8c. Various nuclear DNA patterns were observed in the histologic samples; a recurrent lesion was later found in an area adjacent to the primary lesions that had higher ploidy levels. Ultrastructurally, the Paget cells contained lysosomes and tiny electron-dense secretory granules in their cytoplasms, suggestive of a glandular cell origin. These findings suggest that Paget cells may be derived from the secretory portion of sweat glands. PMID- 2557853 TI - Relationship between computerized morphonuclear image analysis and histopathologic grading of breast cancer. AB - A pilot study analyzed the relationship between several morphonuclear parameters and the Bloom-Richardson score for 37 invasive, not-otherwise-specified (NOS) ductal breast carcinomas. The SAM-BA 200 cell image processor and its software were used to measure the nuclear features on Feulgen-stained imprint smears. Two parameters representing the numbers of large and dense chromatin clots and two parameters describing the heterogeneity of the chromatin among nuclei in a specimen evolved in a continuous manner parallel with the Bloom-Richardson score from stages NOS-4 to NOS-8. The systematic measurement of these four parameters on a large series of breast cancers may be able to define an objective and reproducible "scale" of differentiation that could be a helpful tool for pathologists and clinicians. PMID- 2557854 TI - Small cell undifferentiated carcinoma of the maxillary sinus: technical considerations for radiation therapy. AB - Treatment principles for extrapulmonary small cell undifferentiated carcinoma follow those established for primary lung presentations including systemic chemotherapy and prophylactic cranial irradiation. Radiation of carcinoma involving the maxillary sinus includes lateral and anterior portals which exit through the brain. Presented is a treatment strategy used for postoperative radiation of small cell undifferentiated carcinoma of the maxillary sinus in conjunction with prophylactic cranial irradiation. Specifics of dosimetry include achievement of 55 to 60 Gy within the tumor volume of the maxillary sinus, while achieving homogeneity of dose (30 Gy) within the cranial contents. Application of the described technique to analogous clinical presentations is also discussed. PMID- 2557855 TI - DNA damage stimulates human cell transformation by integrative but not episomal Epstein-Barr virus-derived plasmid. AB - Previous work has demonstrated that ultraviolet (UV) irradiation of SV40-based plasmids can strikingly enhance the frequency of stable transformation of human cells. In this study we compared the effect of UV-induced DNA damage on transformation mediated by integrative versus autonomously replicating plasmids derived from human Epstein-Barr virus (EBV). We report that transfection of human fibroblasts with UV-irradiated integrative EBV-based plasmid results in enhanced transformation. However, transfection of UV-damaged episomal EBV-based constructs into the same human cell line does not enhance transformation; in fact, the extrachromosomal status of the plasmid is maintained irrespective of the UV dose to the plasmid. We conclude that enhanced transformation of human cells by damaged DNA requires its chromosomal integration. PMID- 2557857 TI - [The effect of a submaximal physical load on the beta 2-adrenoreceptor-dependent adenylate cyclase system of the lymphocytes in hypertension]. AB - Effects of dynamic exercise on lymphocyte beta 2-adrenoreceptor density (BARD) and adenylate cyclase activity (ACA) basal and stimulated by isoproterenol (I). Gpp(NH)p, Gpp(NH)p+I, and forskolin (F), on plasma adrenaline and noradrenaline levels, renin activity (PRA) were compared in 6 healthy donors and 12 patients with essential hypertension (EH). Acute stimulation of sympathetic activity by dynamic exercise leads to a rapid increase in lymphocyte BARD in normotensive and hypertensive subjects. The rise in BARD was accompanied by a significant increase in lymphocyte basal ACA in normotensive subjects, but not hypertensive patients. In patients with EH, basal ACA changed after exercise according to their renin status: patients with low baseline PRA showed a decrease in basal ACA after exercise, whereas patients with normal baseline PRA showed an increase as did normotensive subjects. Patients with decreased basal ACA after exercise also showed a lower ACA stimulation by IPR, F, Gpp (NH)p and Gpp(NH)p+IPR, which did not change after exercise. These patients also exhibited substantial rises in PRA with exercise. These was an inverse relationship between exercise-induced changes in PRA and basal ACA. So the acute regulation of lymphocyte BARD-ACA system is changed in a subset of patients with EH. The pattern of the regulation of this lymphocyte system is associated that of baseline renin activity and of its response to exercise in patients with EH. PMID- 2557856 TI - Identification of a transformation-sensitive nuclear protein from normal human fibroblasts that specifically interacts with minute virus of mice DNA and correlates with cell resistance to the parvovirus. AB - Normal human fibroblasts (MRC-5, KMS-6) were compared to transformed derivatives induced by SV40 (MRC-5V1) or gamma rays (KMST-6) for the expression of nuclear proteins that interact with the genome of minute virus of mice (MVMp), using the southwestern blot technique. A protein of 100-104 kDa apparent molecular weight was found to form a specific complex with MVMp DNA and to have an especially high affinity for the 3' terminal portion of the viral genome. This protein (p102) was differentially expressed by normal and transformed cells, i.e., its availability or DNA binding activity (or both) was much reduced in the transformants. A high level of p102 cosegregated with resistance to MVMp in cell hybrids between normal human and transformed mouse fibroblasts. Taken altogether these data suggest that the p102 protein may be a candidate for a transformation-sensitive cellular marker and for a negative regulator of parvovirus replication. PMID- 2557858 TI - [Changes in the characteristics of the lymphocyte beta 2-adrenoreceptors under the influence of prostaglandin E2 infusions in hypertension patients]. AB - Lymphocyte beta 2-adrenoreceptor density and affinity for catecholamines (CA) were examined in 22 patients with essential hypertension (EH) which was resistant to therapy. Lymphocyte beta 2-adrenoreceptor density was 1.5 times more in EH patients than in normotensive donors, but the receptor affinity for CA was the same. Prostaglandin E2 infusions permitted the resistance to antihypertensive therapy be overcome in 17 patients; it was accompanied by a decrease in of lymphocyte beta 2-adrenoreceptor density, receptor's affinity for 1-isoproterenol showed a 7-fold increase. There was no steady antihypertensive effect on prostaglandin E2 infusions in 5 EH patients: in these cases lymphocyte beta 2 adrenoreceptor density increased and their affinity for CA decreased. PMID- 2557860 TI - [The diagnostic importance of single-photon emission-computed tomography in determining the location and extent of acute myocardial infarct]. PMID- 2557859 TI - [The amount of thrombocyte alpha 2-adrenoreceptors in patients with different clinical variants in the course of ischemic heart disease]. AB - Number of alpha-2-adrenoceptors on the thrombocyte membranes was measured with selective antagonists [H]3 yohimbine in 25 patients with ischemic heart disease (7 patients with unstable 12 with vasospastic and 6 with new onset angina pectoris) and in 16 patients without ischemic heart disease and coronary atherosclerosis (control group). The number of alpha-2-adrenoceptors in patients with unstable angina pectoris (571.0 +/- 92.9) was proved to exceed significantly that in patients with vasospastic (237.9 +/- 30.0) or new onset angina (126.2 +/- 19.3) and in the control group (200.0 +/- 22.5). The number of adrenoceptors tended to decrease during myocardial ischemia. PMID- 2557861 TI - High-speed DNA sequencing: an approach based upon fluorescence detection of single molecules. AB - We are developing a laser based technique for the rapid sequencing of large fragments (approximately 40 kb) of DNA based upon the detection of single, fluorescently tagged nucleotides cleaved from a single DNA fragment. We have demonstrated significant progress on several of the important steps of this technique. The projected rate of sequencing is several hundred bases per second which is orders of magnitude faster than existing methods. Once developed, this technology could be utilized by investigators for rapid sequencing of genetic material from virtually any source. PMID- 2557862 TI - Role of the environment in the interaction of nonintercalators with Z-DNA. AB - Interaction of the DNA binding nonintercalators Netropsin, Distamycin and the mPD derivative with Z-DNA has been studied. It has been found that environmental factors like the solvent and added cations significantly modulate the interaction of these ligands with Z-DNA. However no definite Z to B transition in presence of these ligands was found in any case, in contrast to previously reported results (Ch. Zimmer, C. Marck and W. Guschlbauer, FEBS Lett. 154, 156-160 (1983)). PMID- 2557863 TI - Existence of the C structure in poly(dA-dC).poly(dG-dT). AB - We show that the lithium salt of calf-thymus DNA can assume the C structure in nonoriented, hydrated gels. The transitions between the B and C structures showed little hysteresis and none of the metastable structural states which occur in oriented gels. Therefore crystal-lattice forces are not needed to stabilize the C structure. The occurrence of the alternative structures of the Li, Na and K salts of poly(dA-dC).poly(dG-dT) was measured as a function of hydration for nonoriented gels. Poly(dA-dC).poly(dG-dT).Li exists in the B structure at high hydrations and in the C structure at moderate hydrations with no A or Z structure at any hydration tested. The Na salt of poly(dA-dC).poly(dG-dT) exists in the B structure at high hydration, as mixtures of B and C at moderate hydrations and in the A structure at lower hydrations. The potassium salt behaves similarly except that mixtures of the C and A structures exist at lower hydrations. ZnCl2 and NaNO3, which promote the Z structure in duplex poly(dG-dC), promote the C structure in poly(dA-dC).poly(dG-dT). Information contained in the sequence of base pairs and not specific ionic interactions appear to determine the stability of the alternative structures of polynucleotides as hydration is changed. PMID- 2557864 TI - Altered susceptibility to viral respiratory infection during short-term exposure to nitrogen dioxide. AB - The studies reported here focus on the relation of nitrogen dioxide exposure to susceptibility to viral respiratory infection in a murine model of pneumonia, created by intratracheal inoculation of an endogenous murine pathogen, mouse cytomegalovirus. The purpose of this work is to clarify the potential role of nitrogen dioxide exposure in the pathogenesis of viral infection of the lower respiratory tract. Previous human epidemiologic studies have presented conflicting information about the relationship of nitrogen dioxide to acute, self limited episodes of respiratory illness, which are characteristic of viral respiratory infection. Some studies have found an association between exposure to elevated ambient levels of nitrogen dioxide and increased occurrence of acute respiratory illness. In one study this association was found to be strongest in children in the first two years of life. However, other epidemiologic studies have failed to observe this relation. To determine if there is scientific evidence for the possible relation of nitrogen dioxide exposure to human respiratory infection, our studies were performed to assess the impact of nitrogen dioxide on respiratory tract susceptibility to initial, or primary, infection, as well as to recurrent infection, or reinfection, with the identical virus. The latter mechanism of viral respiratory infection is of particular interest, since reinfection is a common method for the development of infection of the lower respiratory tract during early childhood. Outbred CD-1 mice were exposed to either air or nitrogen dioxide for six hours a day on two consecutive days prior to inoculation with murine cytomegalovirus, and then were reexposed to the same level of nitrogen dioxide for six hours a day on four consecutive days, beginning the day after viral inoculation. Susceptibility to primary infection was determined by inoculating animals with an amount of virus (10(2) plaque forming units) that is too small to produce viral infection in the lungs of normal animals. Mice exposed to 5 parts per million (ppm) nitrogen dioxide routinely developed viral replication in the lung and histologic evidence of pneumonitis after inoculation with this amount of virus, whereas air-exposed animals did not. Most importantly, animals exposed to 5 ppm nitrogen dioxide could be infected with a viral inoculum that was 100-fold smaller than that required to consistently produce viral infection in air-exposed mice. Enhanced susceptibility to infection was found after exposure to 5 ppm nitrogen dioxide, but was not observed with exposure to 2.5 or 1 ppm nitrogen dioxide.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557867 TI - Structure of copper(II) perchlorate hexahydrate. AB - Cu(ClO4)2.6H2O, Mr = 370.54, monoclinic, P2(1)/c, a = 5.137 (1), b = 22.991 (3), c = 13.849 (2) A, beta = 90.66 (1) degree, V = 1635.4 (4) A3, Z = 6, Dx = 2.26 g cm-3, lambda(Mo K alpha) = 0.71069 A, mu = 26.44 cm-1, T = 296 K, F(000) = 1122, 2911 unique reflections having I greater than sigma 1, R = 0.030. Each of the two inequivalent copper ions is coordinated by six water-molecule O atoms in a significantly distorted octahedral arrangement. Each of the three inequivalent but geometrically quite similar perchlorate groups is slightly distorted from regular tetrahedral geometry. The overall observed mean Cl-O distance is 1.429 (5) A while the overall observed mean perchlorate O-O distance is 2.333 (8) A. Both the Cu-O complexes and the perchlorate ions were tested and found to behave as rigid bodies. The perchlorate-ion parameters corrected for rigid-body motion are: overall mean Cl-O distance, 1.453 (6) A; overall mean perchlorate O-O distance, 2.372 (8) A. Location and refinement of the 18 H atoms gave a detailed account of hydrogen bonding, which occurs between oxygen octahedra, between oxygen octahedra and perchlorate groups, and, more weakly, within oxygen octahedra. PMID- 2557865 TI - Early markers of lung injury. AB - The goal of this study was to develop an early marker of lung injury that might change in response to exposure to a mobile source emission. Nitrogen dioxide (NO2)2 was chosen as an example of an atmospheric pollutant that is related to automobile emissions. Since reorganization of the connective tissue matrix of the lung occurs in response to injury, markers of connective tissue metabolism were selected as targets. Hydroxylysine became the marker of choice. It is an amino acid that is virtually exclusive to collagen, although it does occur in minimal amounts in other proteins. Furthermore, it is excreted in the urine, which makes it readily available for analysis using noninvasive techniques. Other markers evaluated as part of the study included angiotensin-converting enzyme as a marker of lung injury, desmosine as a marker of elastin degradation, and hydroxyproline as another marker of collagen metabolism. Male Fischer-344 rats were exposed in whole-body chambers to controlled concentrations of NO2 for various doses and periods of time. The concentrations of NO2 ranged from 0.5 to 30 parts per million (ppm); the rats were exposed for six hours per day for periods of two days to four weeks. Urine and bronchoalveolar lavage samples were collected and analyzed for the appropriate marker. In addition, pulmonary function studies and histologic examinations of the lungs were completed at selected time points. Urinary hydroxylysine concentration increased as a function of NO2 concentration during six-hour-per-day exposures for two days. This short-term exposure required relatively high doses to achieve significant changes in the hydroxylysine output. During one-week exposures to either 25 or 30 ppm NO2, there was an increase in urinary hydroxylysine associated with changes in lavage concentrations of angiotensin-converting enzyme and hydroxylysine. The lungs of these animals demonstrated histologic changes typical of oxidant injury. Four-week exposure protocols using 0.5 and 1 ppm NO2 were most interesting in terms of the sensitivity of the marker. There was minimal damage revealed by the histology and function studies, yet there were significant increases in the excretion of hydroxylysine. It appears that hydroxylysine can be indicative of exposure when other parameters are normal. It will require long-term follow-up of exposed rats to determine whether or not the change in marker concentration is predictive of damage. Hydroxylysine may be an excellent marker of exposure to oxidants in the human population. Controlled studies to establish base-line values are needed, followed by carefully controlled studies in individuals with connective tissue abnormalities of the lung. PMID- 2557868 TI - Fluoride uptake from NaF/pyrophosphate anticalculus dentifrices in vitro: effect of pyrophosphate dose. AB - Fluoride uptake from anticalculus dentifrices containing combinations of sodium fluoride and variable doses of soluble pyrophosphate has been evaluated in an in vitro fluoride uptake model. Artificial carious lesions were treated for 30 minutes in 25% w/w saliva slurries of dentifrices. Fluoride uptake into the partially demineralized enamel was assessed by microdrill biopsy. Results show equivalent fluoride uptake for dentifrices containing 0.243% sodium fluoride in combination with 0, 4.0 and 5.0% soluble pyrophosphate. These results, coupled with analytical measures of available fluoride, suggest no negative effects of pyrophosphate on fluoride availability or access to artificial carious lesions for soluble pyrophosphate concentrations up to 5.0% in a conventional (1,100 ppm fluoride) sodium fluoride dentifrice system. PMID- 2557866 TI - Structure of a kappa-opioid receptor misfit: (1S,5R,8R,9R)-2'-hydroxy-5,9 dimethyl-8,2-epoxyethano-6,7-benzomorphan hydrochloride. AB - C16H22NO+2.Cl-, Mr = 295.808, monoclinic, P2(1), a = 11.967 (1), b = 12.529 (1), c = 9.9369 (9) A, beta = 93.00 (1) degrees, V = 1487.8 (2) A3, Z = 4, Dm = 1.32 (2), Dx = 1.321 Mg m-3, lambda(Cu K alpha) = 1.54178 A, mu(Cu K alpha) = 2.289 mm 1, F(000) = 632, T = 291 K, final R = 0.040 for 2448 observed reflections. The two molecules present in the asymmetric unit are linked by an extensive network of hydrogen bonds, including several of the less common (C-)H...O and (C-)-H...Cl types. This interpretation is substantiated by a Mulliken population analysis resulting from CNDO/2 calculations. The major effect of the presence of the epoxyethano bridge is a marked flattening about the N atom of the piperidinium ring. Whether this is sufficient to explain the inactivity of the compound at the opioid kappa receptor is not clear. PMID- 2557869 TI - Clinical comparison of the anticalculus effect of two mouthrinses. AB - A three-month, double-blind clinical study was conducted to compare the effects of supragingival calculus deposits of a mouthrinse containing soluble pyrophosphate and a copolymer of methoxyethylene and maleic acid, as compared to a mouthrinse containing only soluble pyrophosphate and a placebo mouthrinse. Male and female adult subjects were stratified into three balanced groups according to baseline calculus scores obtained from a pre-test period. They received an oral prophylaxis and were assigned to the use of one of the three test mouthrinses. All subjects used a commercially available dentifrice containing 0.76% sodium monofluorophosphate in a silica base. The results from the three-month calculus examination indicated that the soluble pyrophosphate/copolymer mouthrinse reduced supragingival deposits by 31.70% (P less than 0.03), as compared to the placebo mouthrinse. The mouthrinse containing only soluble pyrophosphate did not provide a statistically significant reduction in calculus deposits, as compared to the placebo mouthrinse. PMID- 2557870 TI - [Choice for the treatment of congestive heart failure]. PMID- 2557871 TI - The platelet alpha 2-adrenoceptor as a potential biological marker in depression. PMID- 2557872 TI - [Histological study of malignant cerebral granular cell tumor]. AB - Granular cell tumor (GCT), which is suspected to be of Schwann cell origin, sometimes grows in the subcutaneous tissue, oral cavity and visceral sites and this tumor has a rather benign nature. Intracranial GCT also grows in the neurohypophysis but rarely in the brain parenchyma. We reported a case of intra cerebral GCT in the left hemisphere, which took a malignant course. The patient was a 62-year-old male with a history of slowly progressing right hemiparesis and aphasia since May 1986. He was in a drowsy state and showed right hemiplegia on admission (October 14, 1986). Radiological examinations revealed a tumor and surrounding edema in the left temporal lobe and basal ganglia . Resection of the tumor and both radiotherapy of 53 Grey and chemotherapy using ACNU (total 310 mg) and BrdU (500 mg, two times per week prior to radiation) were applied after the operation. Although the tumor disappeared once after these treatments, the patient died of recurrence on July 3, 1987. Histological examinations on the specimen taken at the first operation revealed that the tumor consisted of rather round, large and small cells with a few cell processes. The large cells often had bizarre and multiple nuclei. These large cells had rich eosinophilic granular particles of various size and vacuoles in their cytoplasm. The staining for antiglial fibrillary acidic protein (GFAP) was positive in a part of the cytoplasm and cell processes. Electron microscopically various sized and shaped granular structures and intermediate filaments were noticed in the cytoplasm of both large and smaller cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557873 TI - [Parkinson's disease: the distribution of Lewy bodies in the peripheral autonomic nervous system]. AB - The distribution of Lewy bodies (LBs) in the peripheral autonomic nervous system was examined in the following 3 groups: group A, 10 patients with Parkinson's disease (age range, 56-82 years); group B, 5 nonparkinsonian patients with many LBs in the central nervous system (CNS) (range, 26-79 years); group C, 176 nonparkinsonian patients without LBs in the CNS (range, 7-107 years). In group A, LBs were found in the paravertebral and/or celiac sympathetic ganglia in 9 cases, enteric nervous system in all cases, cardiac and pelvic plexuses in 4 cases each, and adrenal medullae in 3 cases. They were almost exclusively intraneural . LBs were also found in group B: sympathetic ganglia in 4 cases, enteric nervous system in 5 cases, and pelvic plexus and adrenal medullae in one case each. Interestingly, both the distribution and the number of LBs in a patient with diffuse Lewy body disease were similar to those in group A. LBs, although definitely fewer in number, were also found in group C: sympathetic ganglia in 5 out of 136 cases; enteric nervous system in 8 out of 40 cases; and cardiac plexus in 2 out of 25 cases. All of these positive cases were over age 60. The wide occurrence of LBs in the peripheral autonomic nervous system in patients with Parkinson's disease may play an important role in causing a variety of autonomic symptoms in the disease. On the other hand, LBs have been occasionally found in nonparkinsonian patients over age 60.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557874 TI - Procollagenase associated with the noncalcified matrix of bone and its regulation by parathyroid hormone. AB - Collagenolytic enzyme activity associated with the noncalcified pool of collagen was studied using calvarial matrices from which the periosteal cell envelope had been removed. Aminophenylmercuric acetate (APMA) stimulated degradation of about 5% of the noncalcified collagen in matrices prepared from freshly dissected bone. Significantly more activity was detected if intact calvaria were cultured 24 h before removal of the cells, in which case 20-30% of the noncalcified collagen was degraded following treatment with APMA. Trypsin elicited a similar response. The collagen being degraded was representative of the entire pool of noncalcified collagen and was not underhydroxylated. Treatment of intact calvaria with parathyroid hormone (PTH) before removal of the cells increased the level of both active collagenase and procollagenase activity associated with the matrix. Enhanced 3H release was noted for PTH treated intact bone in the prior 24 h. Inactivation of endogenous procollagenase by phenanthroline had no effect on the ability of isolated calvarial cells to resorb the bone upon treatment with resorptive agents. The data show that PTH-stimulated collagenolysis of noncalcified collagen involves increased deposition of procollagenase onto the noncalcified matrix in addition to activation of the enzyme. PMID- 2557875 TI - Inhalation anaesthetics block accommodation of pyramidal cell discharge in the rat hippocampus. AB - The effects of the three inhalation anaesthetics enflurane, isoflurane and halothane were tested in vitro on accommodation of rat CA1 neurones. At near clinical concentrations (approximately 2.5%) the anaesthetics slightly depressed antidromic field potential responses. At the same concentrations the anaesthetics also blocked accommodation reversibly and reduced the after hyperpolarization of CA1 neurones. No significant changes in the threshold potential were observed, although the resting membrane potential was often increased in the presence of the anaesthetics. The action of enflurane was not blocked by propranolol 20 mumol litre-1 and enflurane had no obvious effect on the duration of Ca2+ spikes of CA1 neurones. It is concluded that the anaesthetics may have a direct effect on membrane K+ channels such as the Ca2+-activated K+ conductance and that the block of accommodation is unlikely to account for the proconvulsant action of enflurane. PMID- 2557878 TI - Treatment of asthma in adults. PMID- 2557877 TI - Effects of antihypertensive agents on endothelium-dependent and endothelium independent relaxations. AB - 1. Antihypertensive agents normalize blood pressure and restore depressed endothelium-dependent relaxations in experimental models of hypertension, but little is known regarding whether antihypertensive agents themselves can directly modulate responses to agonists of endothelium-dependent or independent relaxations, or contractions. 2. Normal rats were treated with either tap water, captopril, hydralazine or enalapril in their drinking water for 2 weeks, following which endothelium-dependent and endothelium-independent relaxations were tested with acetylcholine and sodium nitroprusside, respectively, in aortic rings suspended in organ chambers. 3. All antihypertensive agents caused slight but similar potentiation of sodium nitroprusside-induced relaxations. However, their effects on acetylcholine-induced relaxations were quite different: captopril had a marked potentiating effect, hydrazaline a slight potentiating effect, and enalapril had no significant effect on these relaxations. 4. The relaxations induced by acetylcholine and potentiated by captopril were not altered when indomethacin was included in the tissue bath. However, pyrogallol, an inhibitor of endothelium-derived relaxing factor (EDRF), markedly inhibited these relaxations suggesting that captopril's effect may involve EDRF. 5. SQ 14,534, a stereoisomer of captopril which is 100 fold less potent in inhibiting angiotensin converting enzyme, also significantly enhanced acetylcholine induced relaxations. Thus the effects of both captopril and SQ 14,534 upon EDRF appear independent of the effects of these compounds on the angiotensin converting enzyme. 6. We conclude that certain antihypertensive agents may modulate endothelium-dependent relaxations in response to agonists, and that these properties may be of therapeutic importance in cardiovascular diseases. PMID- 2557876 TI - Comparisons in vitro, ex vivo, and in vivo of the actions of seven structurally diverse inhibitors of angiotensin converting enzyme (ACE). AB - 1. Seven drugs (captopril, zofenopril, enalapril, ramipril, lisinopril, fosinopril, and SQ 29,852) were compared in vitro in homogenates of aorta, brain, heart, lung, and kidney and in sera of spontaneously hypertensive rats (SHR) both with respect to potencies of their active moieties as inhibitors of angiotensin converting enzyme (ACE), and, where applicable, rates of hydrolysis of their prodrug ester functions. 2. In ex vivo dose-response and time-course studies, the inhibitory effects of the seven drugs on tissue ACEs and their relative distributions to SHR tissues were compared following oral administration. 3. The relative potencies of the inhibitory moieties of the drugs (in parentheses) and the normalized 'equiactive' oral doses employed for time-course studies were: SQ 29,852 (1.0), 100 mg kg-1; captopril (3.5), 30 mg kg-1; enalapril (12), 20 mg kg 1; fosinopril (13), 25 mg kg-1; zofenopril (20), 10 mg kg-1; lisinopril (24), 10 mg kg-1; and ramipril (51), 5 mg kg-1. 4. Following oral administration of the drugs to SHR, the degree and duration of ACE inhibition in aorta and lung correlated with the antihypertensive actions, with ramipril, lisinopril, and zofenopril producing effects of the greatest magnitude and duration. 5. Ramipril and enalapril did not inhibit brain ACE ex vivo; captopril and zofenopril had modest but short-lasting effects; and fosinopril, lisinopril, and SQ 29,852 had long-lasting inhibitory actions, which, with the latter two, were delayed in onset. 6. All of the drugs produced significant inhibition of kidney ACE, with ramipril and fosinopril having somewhat weaker effects, perhaps due to biliary routes of excretion. 7. Captopril, fosinopril, and particularly zofenopril inhibited cardiac ACE ex vivo with degrees and durations that were marked compared with those of the other drugs; preliminary studies with isolated hearts suggest a possible relationship between inhibition of cardiac ACE and preservation of cardiac function subsequent to ischaemia. PMID- 2557879 TI - Tissue expression of the tumour associated antigen CA242 in benign and malignant pancreatic lesions. A comparison with CA 50 and CA 19-9. AB - The expression of a novel tumour associated antigen CA 242, defined by the monoclonal antibody C 242, was studied by immunoperoxidase staining in formalin fixed, paraffin-embedded tissue sections from normal pancreata, pancreata with pancreatitis and benign and malignant pancreatic neoplasms. The antigenic determinant of the C 242 antibody is a sialylated carbohydrate structure, related but chemically different from tumour marker antigens CA 19-9 and CA 50. Thirty eight of 41 (93%) well to moderately differentiated ductal adenocarcinomas of the pancreas and all cystadenocarcinomas were positive for CA 242. The staining was most intense in the apical border of the cells, and in the intraluminal mucus. Only two out of seven poorly differentiated adenocarcinomas stained, and the number of positive cells was smaller than in well differentiated carcinomas. Only occasional cells were stained in one out of five anaplastic carcinomas. Part of large ducts were positive in 91% (21/23) specimens of chronic pancreatitis. In acute pancreatitis small terminal ducts, centro-acinar cells and some large ducts stained for CA 242. In normal pancreas only a few small terminal ducts were CA 242 positive. Carcinomas always stained more strongly for CA 242 than normal pancreatic tissue adjacent to the carcinoma. The results of CA 242 are compared with those of tumour marker antigens CA 50 and CA 19-9. Serum CA 242 levels were determined in 23 of the patients with pancreatic cancer using a fluoroimmunoassay. Fifteen (65%) patients had an elevated value. There was no clear-cut correlation between the serum levels and the immunohistochemical expression of the CA 242 antigen. The expression of CA 242 in pancreatic tissue resembles that of CA 50 and is similar to CA 19-9. The antigen is expressed in serum of many patients with pancreatic cancer and, therefore, is a potential candidate for a serum tumour marker. PMID- 2557881 TI - Recombinant alpha 2 interferon is superior to doxorubicin for inoperable hepatocellular carcinoma: a prospective randomised trial. AB - In a prospective trial of 75 Chinese patients with histologically proven inoperable hepatocellular carcinoma (HCC), 25 patients were randomised to receive doxorubicin 60-75 mg m-2 intravenously once every 3 weeks, 25 to receive recombinant alpha 2 interferon (rIFN) (Roferon) 9-18 x 10(6) IU m-2 intramuscularly (i.m.) daily and 25 to receive rIFN 25-50 x 10(6) IU m-2 i.m. three times weekly. Patients were switched to the other drug if: (a) there was progressive disease after 12 weeks, (b) unacceptable toxicity developed and (c) they had received a total of 500 mg m-2 of doxorubicin. Six patients had switching over of therapy, three on doxorubicin and three on rIFN. In the remaining 69 patients on single drug therapy, the median survival rate of patients on doxorubicin and rIFN was 4.8 and 8.3 weeks respectively (P = ns.). rIFN induced tumour regression of 25-50% in 12% of patients and of over 50% in 10% of patients. When compared with doxorubicin, rIFN was associated with more tumour regression (P = 0.00199) and less progressive tumours (P = 0.00017). It caused less prolonged and less severe marrow suppression (P = 0.01217), and had significantly less fatal complications than doxorubicin (P = 0.01383). Doxorubicin caused fatal complications due to cardiotoxicity and neutropenia in 25% of patients. rIFN was associated with fatal complications due to dementia and renal failure in 3.8% of patients. In the treatment of inoperable HCC, rIFN is superior to doxorubicin in causing more tumour regression, less serious marrow suppression and less fatal complications. PMID- 2557880 TI - Characterisation of human thyroid epithelial cells immortalised in vitro by simian virus 40 DNA transfection. AB - Human primary thyroid follicular epithelial cells were transfected with a plasmid containing an origin-defective SV40 genome (SVori-) to produce several immortal cell lines. Two of the 10 cell lines analysed expressed specific features of thyroid epithelial function (iodide-trapping and thyroglobulin production). These two lines were characterised in detail and found to be growth factor-independent, capable of anchorage-independent growth at low frequency but non-tumorigenic in nude mice. These differentiated, These differentiated, partially transformed cell lines were shown to be suitable for gene transfer at high frequency using simple coprecipitation techniques. PMID- 2557882 TI - Amelioration of established Sendai viral pneumonia in the nude mouse using a monoclonal antibody to the virus fusion protein. AB - The pathological effect of parainfluenza type I (Sendai virus) is known to be a bronchopneumonia, which becomes a chronic pneumonia in the immunodeficient athymic (nude) mouse. The severity of this established chronic pneumonia can be dramatically altered by providing the nude mouse with humoral monoclonal antibodies which are neutralizing, and are directed against the fusion protein, of the virus. The alveolitis, which is a significant part of the pathology, is suppressed due to a reduction (greater than 90%) in the number of virus-infected alveolar macrophages present in the alveoli. This clearly identifies the infected alveolar macrophage as the primary effector cell in the pathogenesis of alveolitis caused by parainfluenza virus type I. The implications of using virus neutralizing monoclonal antibodies, which have little immunomodulatory toxicity, in the treatment of viral pneumonias are discussed. PMID- 2557884 TI - A spectrophotometric method for the assay of pyrimidine 5'-nucleotidase in human erythrocytes. AB - A new spectrophotometric assay for human erythrocyte pyrimidine 5'-nucleotidase using a continuous spectrophotometric method is described. The activity is detected by measuring uracil, which is produced by a coupled assay including cytidine deaminase and uridine nucleosidase. The results obtained with the continuous spectrophotometric method have been confirmed by high pressure liquid chromatography (HPLC) analysis. This procedure, not based upon inorganic phosphate formation and avoiding the use of radioactive material, offers significant advantages with respect to commonly adopted assays. PMID- 2557885 TI - Aplastic anaemia following successful treatment of malignant epithelial tumours with radiation and/or chemotherapy. PMID- 2557883 TI - Free radicals, reactive oxygen species and human disease: a critical evaluation with special reference to atherosclerosis. PMID- 2557887 TI - Influence of post-rumen supply of nutrients on rumen digesta load and voluntary intake of a roughage by sheep. AB - The effects of post-rumen supply of nutrients on the rumen digesta load and voluntary consumption of roughage by Border Leicester X Merino ram lambs was investigated. A chopped wheaten hay was offered to the ram lambs (n 24), whose nutrient intake was altered by infusing into the abomasum a liquid supplement, containing reconstituted cow's milk, sodium caseinate, minerals and vitamins, for a period of at least 30 d. Either two or three lambs were allocated by live weight to each of ten rates of nutrient supplementation. The lambs were slaughtered at a target live weight of 31 kg. Voluntary intake of hay was estimated over the last 7 d of feeding. The amounts of digesta and organic matter (OM) in the rumen were measured by emptying after slaughter. The particle size distribution of the digesta was measured by wet sieving and the fractional outflow rate of particulate matter by reference to the marker lignin. Growth rate of the lambs increased linearly as total energy infused increased. The linear increase in growth rate indicated that roughage was not substituted for infused nutrients on a direct energy basis. OM intake from hay declined linearly as the amount of nutrients infused increased. The decline in intake was associated with a decline in both total digesta and the amount of rumen OM. Both the fractional digestion rate of OM and the fractional outflow rate of particulate matter from the rumen were unaffected by the amount of nutrients infused. The results indicated that the rumen digesta load of young sheep fed on a single roughage is directly related to their energy deficit, that is the difference between the capacity of the animals to use energy and the energy available to them for metabolism. PMID- 2557886 TI - Digestion of fibre polysaccharides of pea (Pisum sativum) hulls, carrot and cabbage by adult cockerels. AB - Characterization of the carbohydrates of pea (Pisum sativum) hulls, carrot and cabbage using both colorimetric and gas-liquid chromatographic techniques permitted a detailed investigation into the extent of digestion of differing types of fibre. These digestion studies were greatly aided by the development of a rapid bioassay employing starved adult cockerels. Total collection of undigested residues, uncontaminated by food spillage, could be made from trays placed under the cockerels. Chemical analysis showed that pea hulls consisted mainly of fibre with very little available carbohydrate present, whereas more than half of freeze-dried carrot and cabbage consisted of available carbohydrate (sucrose, glucose, fructose, starch) and consequently considerably less fibre was present. The fibre of carrot and cabbage was similarly composed of nearly equal amounts of neutral and acidic polysaccharides, whereas pea-hull fibre had four times as much neutral as acidic polysaccharides. The digestibility of total neutral polysaccharides from all three foodstuffs was extremely low. However, there appeared to be preferential digestion of polysaccharides composed of rhamnose, arabinose and galactose residues, all associated with pectic material, in contrast to the indigestibility of polysaccharides composed of fucose, xylose and glucose. Acidic polysaccharides were digested to a greater extent than neutral ones, and those of carrot and cabbage more so than pea hulls. The polysaccharides which were the most soluble were also the most digestible, but due to the arbitrariness of polysaccharide solubility, quantification of their total digestibility per se was considered not possible. PMID- 2557888 TI - Energy expenditure associated with sodium/potassium transport and protein synthesis in skeletal muscle and isolated hepatocytes from hyperthyroid sheep. AB - The object of the present study was to determine the effect of thyroxine (T4) treatment of sheep on protein synthesis and associated energy costs in skeletal muscle and hepatocytes. Protein synthesis, and ouabain-sensitive and cycloheximide-sensitive respiration in isolated intercostal muscle and hepatocytes were determined in sheep after 5 weeks of daily injections of either saline or T4. Plasma T4 and total triiodothyronine (T3) concentrations were doubled and free T3 concentrations were quadrupled by T4 injections. The fractional rates of protein synthesis increased in isolated external intercostal muscle and hepatocytes from hyperthyroid sheep. Fractional rates of protein synthesis in isolated external intercostal muscle and hepatocytes were linearly correlated with plasma free T3 concentrations. Total oxygen consumption of muscle and hepatocytes was unaffected by T4 injections. Ouabain-sensitive respiration increased in hepatocytes and muscle of T4-treated animals. Cycloheximide sensitive respiration was elevated in hepatocytes from hyperthyroid sheep. Cycloheximide-sensitive respiration in muscle was unaffected by T4 treatment. The present experiment demonstrates that T4 increases protein synthesis in ruminants. The energy expenditure in support of Na+, K(+)-ATPase and protein synthesis in skeletal muscle and hepatocytes may account for 34-60% of total cellular energy expenditure. PMID- 2557889 TI - Breast milk fatty acids in mothers of children with atopic eczema. AB - The total lipid fatty acid composition of mature breast milk has been analysed in a group of twenty-five mothers of children with atopic eczema, and compared with breast milk from twenty-two controls. Total lipids were extracted into chloroform methanol (2:1, v/v) and the methyl esters prepared by alkalicatalysed trans esterification were separated by gas-liquid chromatography and identified by comparison with standard fatty acid methyl esters. Results show that mothers of children with atopic eczema have a significantly greater proportion of linoleic acid, and a smaller proportion of dihomo-gamma-linolenic acid in their total breast milk lipid than the controls. Proportions of total derived fatty acids were similar between groups and there were no differences in the principal saturated and monounsaturated fats. It was concluded that mothers of children with atopic eczema have an abnormal breast-milk fatty acid composition. This supports previous evidence of a defect of conversion of linoleic acid into its long-chain polyunsaturated metabolites in the condition. PMID- 2557890 TI - Direct observation of the enzyme-intermediate complex of 5-enolpyruvylshikimate-3 phosphate synthase by 13C NMR spectroscopy. AB - The interaction of 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase, 4,5 dideoxyshikimate 3-phosphate (ddS3P), and [2-13C]-and [3-13C]phosphoenolpyruvate (PEP) has been examined by 13C NMR spectroscopy. Although no resonances due to a dead-end intermediate complex could be detected, an enzyme active site specific formation of pyruvate was observed. The interaction of EPSP synthase with shikimate 3-phosphate (S3P) and [2-13C]- or [3-13C]PEP has been examined by 13C NMR spectroscopy. With [2-13C]PEP, in addition to the resonances due to [2 13C]PEP and [8-13C]EPSP, new resonances appeared at 164.8, 110.9, and 107.2 ppm. The resonance at 164.8 ppm has been assigned to enzyme-bound EPSP. The resonance at 110.9 ppm has been assigned to C-8 of an enzyme-free tetrahedral intermediate of the sort originally proposed by Levin and Sprinson [Levin, J. G., & Sprinson, D. B. (1964) J. Biol. Chem. 239, 1142-1150] and recently independently observed by Anderson et al. [Anderson, K. S., Sikorski, J. A., Benesi, A. J., & Johnson, K. A. (1988) J. Am. Chem. Soc. 110, 6577-6579]. The resonance at 107.2 ppm has been assigned to an enzyme-bound intermediate whose structure is closely related to that of the tetrahedral intermediate. With [3-13C]PEP, new resonances appeared at 88.9, 26.2, 25.5, and 24.5 ppm. The resonance at 88.9 ppm has been assigned to enzyme-bound EPSP. The resonance at 26.2 ppm, which was found to correlate with 1.48 ppm by isotope-edited multiple quantum coherence 1H NMR spectroscopy, has been assigned to the methyl group 4-hydroxy-4-methylketoglutarate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557891 TI - Detection of an oxyferryl porphyrin pi-cation-radical intermediate in the reaction between hydrogen peroxide and a mutant yeast cytochrome c peroxidase. Evidence for tryptophan-191 involvement in the radical site of compound I. AB - Peroxide oxidation of a mutant cytochrome c peroxidase, in which Trp-191 has been replaced by Phe through site-directed mutagenesis, produces an oxidized intermediate whose stable UV/visible absorption spectrum is very similar to that of compound I of the native yeast enzyme. This spectrum is characteristic of an oxyferryl, Fe(IV), heme. Stopped-flow studies reveal that the reaction between the mutant enzyme and hydrogen peroxide is biphasic with the transient formation of an intermediate whose absorption spectrum is quite distinct from that of either the native ferric enzyme or the final product. Rapid spectral scanning of the intermediate provides a spectrum characteristic of an oxyferryl porphyrin pi cation-radical species. At pH 6, 100 mM ionic strength, and 25 degrees C, the rate constant for formation of the oxyferryl pi-cation radical has a lower limit of 6 X 10(7) M-1 s-1 and the rate of conversion of the transient intermediate to the final oxidized product is 51 +/- 4 s-1. Evidence is presented indicating that Trp-191 either is the site of the radical in CcP compound I or is intimately involved in formation of the radical. PMID- 2557892 TI - Raman/absorption simultaneous measurements for cytochrome oxidase compound A at room temperature with a novel flow apparatus. AB - A novel flow apparatus for continuously producing reaction intermediates of cytochrome oxidase was constructed and applied successfully to observe the transient absorption and resonance Raman spectra in its reaction with oxygen. Time-resolved difference absorption spectra in 500-650-nm region clearly indicated the formation of compound A upon photolysis of the fully reduced CO bound form at 5 degrees C, and at this stage electrons were not transferred from cytochrome c to cytochrome oxidase. However, at the stage of formation of compound B, cytochrome c was oxidized. Resonance Raman spectra of these intermediates measured simultaneously with the absorption spectra are also reported. PMID- 2557893 TI - Characterization of reductant-induced, tryptophan fluorescence changes in cytochrome oxidase. AB - We have measured the steady-state tryptophan fluorescence spectrum of cytochrome oxidase in its oxidized and fully reduced states. Reduction of the oxidized enzyme by sodium dithionite causes an apparent shift in the fluorescence emission maximum from 328 nm, in the oxidized enzyme, to 348 nm, in the reduced enzyme. This spectroscopic change has been observed previously and assigned to a redox linked, conformational change in cytochrome oxidase [Copeland, R. A., Smith, P. A., & Chan, S. I. (1987) Biochemistry 26, 7311-7316]. When dithionite-reduced enzyme sits in an open cuvette, the enzyme returns to the oxidized state, and the fluorescence maximum shifts back to 328 nm. However, the time course of the fluorescence change does not follow the redox state of the enzyme, monitored spectrophotometrically at 445,605, and 820 nm, but follows the disappearance of dithionite, which absorbs at 315 nm. Moreover, when the fluorescence emission spectrum of the dithionite-reduced enzyme is corrected for the absorbance due to dithionite, the fluorescence maximum is found 2 nm blue shifted, relative to that of the oxidized enzyme, at 326 nm. This dithionite-induced, red-shifted steady state tryptophan fluorescence is also seen with the non-heme-containing enzyme carboxypeptidase A. The tryptophan emission spectrum of untreated carboxypeptidase A is at 332 nm, whereas in the presence of dithionite the emission spectrum of carboxypeptidase A is at 350 nm. When corrected for the absorbance of dithionite, the tryptophan emission maximum is at 332 nm. We have also used the photoreductant 3,10-dimethyl-5-deazaisoalloxazine (deazaflavin) to reduce cytochrome oxidase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557894 TI - The axial ligands of heme in cytochromes: a near-infrared magnetic circular dichroism study of yeast cytochromes c, c1, and b and spinach cytochrome f. AB - Room temperature near-infrared magnetic circular dichroism and low-temperature electron paramagnetic resonance measurements have been used to characterize the ligands of the heme iron in mitochondrial cytochromes c, c1, and b and in cytochrome f of the photosynthetic electron transport chain. The MCD data show that methionine is the sixth ligand of the heme of oxidized yeast cytochrome c1; the identify of this residue is inferred to be the single conserved methionine identified from a partial alignment of the available cytochrome c1 amino acid sequences. A different residue, which is most likely lysine, is the sixth heme ligand in oxidized spinach cytochrome f. The data for oxidized yeast cytochrome b are consistent with bis-histidine coordination of both hemes although the possibility that one of the hemes is ligated by histidine and lysine cannot be rigorously excluded. The neutral and alkaline forms of oxidized yeast cytochrome c have spectroscopic properties very similar to those of the horse heart proteins, and thus, by analogy, the sixth ligands are methionine and lysine, respectively. PMID- 2557895 TI - Characterization of the multiple forms of cytochrome b559 in photosystem II. AB - Cytochrome b559 is an essential component of the photosystem II (PSII) protein complex. Its function, which has long been an unsolved puzzle, is likely to be related to the unique ability of PSII to oxidize water. We have used EPR spectroscopy and spectrophotometric redox titrations to probe the structure of cytochrome b559 in PSII samples that have been treated to remove specific components of the complex. The results of these experiments indicate that the low temperature photooxidation of cytochrome b559 does not require the presence of the 17-, 23-, or 33-kDa extrinsic polypeptides or the Mn complex (the active site in water oxidation). We observe a shift in the g value of the EPR signal of cytochrome b559 upon warming a low-temperature photooxidized sample, which presumably reflects a change in conformation to accommodate the oxidized state. At least three redox forms of cytochrome b559 are observed. Untreated PSII membranes contain one high-potential (375 mV) and one intermediate-potential (230 mV) cytochrome b559 per PSII. Thylakoid membranes also appear to contain one high potential and one intermediate-potential cytochrome b559 per PSII, although this measurement is more difficult due to interference from other cytochromes. Removal of the 17- and 23-kDa extrinsic polypeptides from PSII membranes shifts the composition to one intermediate-potential (170 mV) and one low-potential (5 mV) cytochrome b559. This large decrease in potential is accompanied by a very small g-value change (0.04 at gz), indicating that it is the environment and not the ligand field of the heme which changes significantly upon the removal of the 17- and 23-kDa polypeptides. PMID- 2557896 TI - Isolation of a protein fraction from Bordetella pertussis that facilitates entry of the calmodulin-sensitive adenylate cyclase into animal cells. AB - Bordetella pertussis, the pathogen responsible for whooping cough, releases a soluble calmodulin-sensitive adenylate cyclase into its culture medium. Several investigators have shown that the partially purified adenylate cyclase is capable of entering animal cells and elevating intracellular cAMP levels [Confer, D. L., & Eaton, J. W. (1982) Science 217, 948-950; Shattuck, R. L., & Storm, D. R. (1985) Biochemistry 24,6323-6328]. However, the mechanism for entry of the catalytic subunit of the adenylate cyclase into animal cells is unknown. Recently, it was determined that the purified catalytic subunit of the enzyme is unable to enter animal cells [Masure, H. R., Oldenburg, D. J., Donovan, M. G., Shattuck, R. L., & Storm, D. R. (1988) J. Biol. Chem. 263, 6933-6940]. On the basis of these data and other observations, we hypothesized that the culture medium of B. pertussis contains one or more additional polypeptides which facilitate entry of the adenylate cyclase catalytic subunit into animal cells. In this study, we report that a cell-invasive preparation of B. pertussis adenylate cyclase was rendered noninvasive after passage through a wheat germ lectin agarose column. A fraction was eluted from the wheat germ lectin-agarose column with N-acetyl-D-glucosamine. This fraction, when combined with the noninvasive adenylate cyclase, was able to restore the ability of the adenylate cyclase preparation to enter neuroblastoma cells and increase intracellular cAMP levels. Furthermore, the fraction eluted from the wheat germ lectin-agarose column was found to be trypsin and chymotrypsin sensitive, suggesting that this material was proteinaceous.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557897 TI - Biochemical and pharmacological properties of p170 and p180 forms of topoisomerase II. AB - The p170 and p180 forms of topoisomerase II have been compared. The concentration dependence of ATP for catalytic activity of the two forms of the enzyme was identical, and each was equally sensitive to novobiocin. Orthovanadate was found to be a potent inhibitor of catalytic activity of both p170 and p180, with an IC50 value of about 2 microM for each. Under standard reaction conditions, relaxation of supercoiled pBR322 by p180 was highly processive, while p170 performed the same reaction in a distributive manner. The optimal concentration of KCl for catalytic activity of p180 was 20-30 mM higher than that for p170. Comparison of their thermal stability showed that p180 was inactivated at twice the rate of p170. Teniposide and merbarone selectively inhibited catalytic activity of p170, requiring concentrations 3-fold and 8-fold lower, respectively, than those required for equivalent inhibition of p180. Similar selectivity for p170 was seen for teniposide-stimulated DNA cleavage or its inhibition by merbarone. Analysis of sites of DNA cleavage indicated a subset of sites that were either preferred or unique for each of the enzymes. A synthetic oligonucleotide representative of p170 sites selectively inhibited the p170 enzyme. Immunoblotting of p170 and p180 from U937 cells at different stages of proliferation showed that p170 levels declined as the cells reached the plateau phase of growth, while p180 levels were low during rapid proliferation and increased as the growth rate slowed. The data indicate that the p170 and p180 forms of topoisomerase II can be distinguished biochemically, pharmacologically, and by differential cellular regulation. PMID- 2557898 TI - Manganese and calcium requirements for reconstitution of oxygen-evolution activity in manganese-depleted photosystem II membranes. AB - The Mn complex of photosystem II and O2-evolution activity are reconstituted in Mn-depleted photosystem II membranes in a light-dependent process called photoactivation. Recovery of O2-evolution activity requires both Mn2+ and Ca2+ in the photoactivation medium. The Mn2+ and Ca2+ dependences of both the effective rate constant and yield of photoactivation have been determined. A comparison of these data with the predictions of mathematical models for photoactivation leads to the conclusion that photoactivation occurs in two stages. The first stage, photoligation of Mn, requires light and depends primarily on Mn2+. The second stage, binding of Ca2+, is required for expression of O2-evolution activity. This two-stage model affords an excellent fit to the data and provides dissociation constants and binding stoichiometries for Ca2+ and Mn2+. We conclude that one Mn2+ ion is bound and photooxidized in the rate-determining step(s) of photoactivation. On the basis of these results and data already in the literature, the molecular details of the elementary steps in photoactivation are discussed and a mechanism of photoactivation is proposed. PMID- 2557899 TI - Carbonyldiphosphonate, a selective inhibitor of mammalian DNA polymerase delta. AB - Twenty-three pyrophosphate analogues were screened as inhibitors of proliferating cell nuclear antigen independent DNA polymerase delta (pol delta) derived from calf thymus. Carbonyldiphosphonate (COMDP), also known as alpha oxomethylenediphosphonate, inhibited pol delta with a potency (Ki = 1.8 microM) 20 times greater than that displayed for DNA polymerase alpha (pol alpha) derived from the same tissue. Characterization of the mechanism of inhibition of pol delta indicated that COMDP competed with the dNTP specified by the template and was not competitive with the template-primer. In the case of pol alpha, COMDP did not compete with either the dNTP or the polynucleotide substrate. COMDP inhibited the 3'----5' exonuclease activity of pol delta weakly, displaying an IC50 greater than 1 mM. PMID- 2557900 TI - Purification of botrocetin from Bothrops jararaca venom. Analysis of the botrocetin-mediated interaction between von Willebrand factor and the human platelet membrane glycoprotein Ib-IX complex. AB - Interaction of von Willebrand factor (vWF) with its platelet receptor only occurs in vitro in the presence of a modulator such as ristocetin. We have recently confirmed that the human platelet membrane glycoprotein (GP) Ib-IX complex is the receptor involved in the ristocetin-dependent binding of vWF by reconstitution with the purified components [Berndt, M.C., Du, X., & Booth, W.J. (1988) Biochemistry 27, 633-640]. We have now developed a similar solid-phase reconstitution assay using an alternate modulator, botrocetin, for the competitive analysis of functional domains in both vWF and the GP Ib-IX complex. Botrocetin was purified from Bothrops jararaca venom by ammonium sulfate fractionation and subsequent DEAE-cellulose and hydroxylapatite chromatography. The purified protein was a 25-kilodalton (kDa) disulfide-linked dimer with apparent subunit molecular weights of 14,000 and 14,500. Binding studies with immobilized botrocetin demonstrated that botrocetin bound to vWF and to a 52/48 kDa region of vWF that contains the GP Ib binding domain, but not to glycocalicin, a proteolytic fragment of GP Ib that contains the vWF binding site. Binding of 125I-labeled vWF to GP Ib-IX complex coated beads and to platelets was strictly botrocetin-dependent with half-maximal binding at a botrocetin concentration of congruent to 0.27 microM. Botrocetin-dependent binding of vWF was specific, saturable, and comparable to that observed with ristocetin. An anti vWF monoclonal antibody, 3F8, inhibited ristocetin- but not botrocetin-dependent binding of vWF, suggesting the presence of distinct ristocetin and botrocetin modulator sites on vWF. The botrocetin reconstitution assay was at least an order of magnitude more sensitive than the corresponding ristocetin assay for the competitive analysis of functional domains on both vWF and the GP Ib-IX complex and has confirmed the localization of the vWF-binding domain to the 45-kDa N terminal region of GP Ib. PMID- 2557901 TI - Formation of free radical metabolites in the reaction between soybean lipoxygenase and its inhibitors. An ESR study. AB - Recent studies showed that soybean lipoxygenase inhibitors like phenidone and nordihydroguaiaretic acid (NDGA) reduce the catalytically active ferric lipoxygenase to its inactive ferrous form. Addition of 13(S)-hydroperoxy-cis 9,trans-11-octadecadienoic acid (13-HPOD) regenerated the active ferric form. In this paper, it is shown that in such a system the inhibitors are oxidized to free radical metabolites. Incubation of soybean lipoxygenase and linoleic acid with p aminophenol, catechol, hydroquinone, NDGA, or phenidone resulted in the formation of the one-electron oxidation products of these compounds. Free-radical formation depended upon the presence of the lipoxygenase and 13-HPOD. The free radicals were detected by ESR spectroscopy, and their structure was confirmed by analysis of the spectra, using a computer correlation technique. These data support the proposed mechanism for the inhibition of lipoxygenase by phenolic antioxidants. PMID- 2557902 TI - Identification of phosphorylation sites for adenosine 3',5'-cyclic phosphate dependent protein kinase on the voltage-sensitive sodium channel from Electrophorus electricus. AB - The voltage-sensitive sodium channel from the electroplax of Electrophorus electricus is selectively phosphorylated by the catalytic subunit of cyclic-AMP dependent protein kinase (protein kinase A) but not by protein kinase C. Under identical limiting conditions, the protein was phosphorylated 20% as rapidly as the synthetic model substrate kemptamide. A maximum of 1.7 +/- 0.6 equiv of phosphate is incorporated per mole. Phosphoamino acid analysis revealed labeled phosphoserine and phosphothreonine at a constant ratio of 3.3:1. Seven distinct phosphopeptides were identified among tryptic fragments prepared from radiolabeled, affinity-purified protein and resolved by HPLC. The three most rapidly labeled fragments were further purified and sequenced. Four phosphorylated amino acids were identified deriving from three consensus phosphorylation sites. These were serine 6, serine 7, and threonine 17 from the amino terminus and a residue within 47 amino acids of the carboxyl terminus, apparently serine 1776. The alpha-subunits of brain sodium channels, like the electroplax protein, are readily phosphorylated by protein kinase A. However, these are also phosphorylated by protein kinase C and exhibit a markedly different pattern of incorporation. Each of three brain alpha-subunits displays an approximately 200 amino acid segment between homologous repeat domains I and II, which is missing from the electroplax and skeletal muscle proteins [Noda et al. (1986) Nature (London) 320, 188; Kayano et al. (1988) FEBS Lett. 228, 1878; Trimmer et al. (1989) Neuron 3, 33]. Most of the phosphorylation of the brain proteins occurs on a cluster of consensus phosphorylation sites located in this segment. This contrasts with the pattern of highly active sites on the amino and carboxyl termini of the electroplax protein. The detection of seven labeled tryptic phosphopeptides compared to the maximal labeling stoichiometry of approximately 2 suggests that many of the acceptor sites on the protein may be blocked by endogenous phosphorylation. PMID- 2557903 TI - Substrate specificity and variables affecting efficiency of mammalian flavin adenine dinucleotide synthetase. AB - Substrate specificity and product inhibition have been evaluated by using purified rat liver FAD synthetase (ATP:FMN adenylyltransferase, EC 2.7.7.2), obtained by an improved purification protocol with optimized flavin affinity chromatography. FMN analogues studied fall into three general classifications: those with substitution on the pyrimidinoid ring and nitrogen replacement, those with substitution on the benzenoid ring, and those with N(10) side chain modifications. Substitutions on the pyrimidinoid ring and replacement of nitrogens have the greatest influence on binding to enzyme and FAD formation. When the hydrogen-bonding capacity of the NH group at position 3 is blocked or removed by substitution, such FMN analogues do not act as substrates or inhibitors of the enzyme. Substitutions on the benzenoid ring by small groups seem to be tolerated, while larger groups inhibit binding. Length of the N(10) side chain is optimal with five carbons and has greatest affinity for the natural ribityl side chain. Affinity matrices show similar binding characteristics in that the N(3)-(carboxymethyl)riboflavin-agarose does not bind enzyme, while agaroses linked to the flavin N(10) side chain provide varying degrees of purification. The C = O group at position 2, the NH group at position 3, and a five-carbon side chain at the N(10) position seem to be most crucial for flavin substrate binding to enzyme. Nucleoside triphosphates other than ATP do not act as substrates or inhibitors when sufficient Mg2+ is present. Products of the reaction, FAD and PPi, act as inhibitors against both ATP and FMN.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557904 TI - Effects of CapZ, an actin capping protein of muscle, on the polymerization of actin. AB - We have studied the interaction of CapZ, a barbed-end actin capping protein from the Z line of skeletal muscle, with actin. CapZ blocks actin polymerization and depolymerization (i.e., it "caps") at the barbed end with a Kd of approximately 0.5-1 nM or less, measured by three different assays. CapZ inhibits the polymerization of ATP-actin onto filament ends with ATP subunits slightly less than onto ends with ADP subunits, and onto ends with ADP-BeF3- subunits about as much as ends with ADP subunits. No effect of CapZ is seen at the pointed end by measurements either of polymerization from acrosomal processes or of the critical concentration for polymerization at steady state. CapZ has no measureable ability to sever actin filaments in a filament dilution assay. CapZ nucleates actin polymerization at a rate proportional to the first power of the CapZ concentration and the 2.5 power of the actin concentration. No significant binding is observed between CapZ and rhodamine-labeled actin monomers by fluorescence photobleaching recovery. These new experiments are consistent with but do not distinguish between three models for nucleation proposed previously (Cooper & Pollard, 1985). As a prelude to the functional studies, the purification protocol for CapZ was refined to yield 2 mg/kg of chicken breast muscle in 1 week. The activity is stable in solution and can be lyophilized. The native molecular weight is 59,600 +/- 2000 by equilibrium ultracentrifugation, and the extinction coefficient is 1.25 mL mg-1 cm-1 by interference optics. Polymorphism of the alpha and beta subunits has been detected by isoelectric focusing and reverse-phase chromatography. CapZ contains no phosphate (less than 0.1 mol/mol). PMID- 2557906 TI - Photoinduced electron-transfer kinetics of singly labeled ruthenium bis(bipyridine) dicarboxybipyridine cytochrome c derivatives. AB - Cytochrome c derivatives labeled at specific lysine amino groups with ruthenium bis(bipyridine) dicarboxybipyridine [RuII(bpy)2(dcbpy)] were prepared by using the procedure described previously [Pan, L. P., Durham, B., Wolinska, J., & Millett, F. (1988) Biochemistry 27, 7180-7184]. Four additional singly labeled derivatives were purified, bringing the total number to 10. These derivatives have a strong luminescence emission centered at 662 nm arising from the excited state, RuII*. Transient absorption spectroscopy was used to directly measure the rate constants for the photoinduced electron-transfer reaction from RuII* to the ferric heme group (k1) and for the thermal back-reaction from the ferrous heme group to RuIII (k2). The rate constants were found to be k1 = 14 X 10(6) s-1 and k2 = 24 X 10(6) s-1 for the derivative modified at lysine 72, which has a distance of 8-16 A between the ruthenium and heme groups. Similar rate constants were found for the derivatives modified at lysines 13 and 27, which have distances of 6-12 A separating the ruthenium and heme groups. The rate constants were significantly slower for the derivatives modified at lysine 25 (k1 = 1 X 10(6) s-1, k2 = 1.5 X 10(6) s-1) and lysine 7 (k1 = 0.3 X 10(6) s-1, k2 = 0.5 X 10(6) s-1), which have distances of 9-16 A. Transients due to photoinduced electron transfer could not be detected for the remaining derivatives, which have larger distances between the ruthenium and heme groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557905 TI - Free radicals induced by adriamycin-sensitive and adriamycin-resistant cells: a spin-trapping study. AB - The radicals generated by adriamycin-sensitive (CHO-AB) and adriamycin-resistant (CHO-C5) Chinese hamster ovary cells as well as by adriamycin-sensitive and resistant human breast cancer cells (MCF7-WT and MCF7-ADR) have been studied with spin-trapping and ESR spectroscopy. During anoxic exposure to adriamycin (ADR) both pairs of cell lines produced the broad ESR singlet characteristic of ADR semiquinone (AQ.). By use of tris(oxalato)chromate (CrOx) as an extracellular line-broadening agent, the distribution of AQ. between the intra- and extracellular compartments was studied. For cell densities of (1-3) X 10(7) cells/mL, CrOx eliminated most, though not all, of the ESR signal, indicating that the AQ. radicals freely diffuse and partition between the intra- and extracellular compartments proportionally to their respective volumes. Similar behavior was exhibited by all four cell lines studied. Upon introduction of oxygen to anoxic cells in the presence of the spin trap 5,5-dimethylpyrroline N oxide (DMPO), the AQ. signal was replaced by that of the DMPO-OH spin adduct. Metal chelators such as desferrioxamine had no effect on DMPO-OH or AQ. formation. Superoxide dismutase, not catalase, totally eliminated the ESR signal, indicating that DMPO-OH produced by ADR-treated cells originates from superoxide rather than from .OH produced from H2O2. In the presence of CrOx, the DMPO-OH signal was not distinguishable from the background noise, thus excluding any contribution to the signal by intracellular spin adducts.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557907 TI - Two-dimensional 1H and 31P NMR spectra and restrained molecular dynamics structure of an extrahelical adenosine tridecamer oligodeoxyribonucleotide duplex. AB - Assignment of the 1H and 31P NMR spectra of an extrahelical adenosine tridecamer oligodeoxyribonucleotide duplex, d(CGCAGAATTCGCG)2, has been made by two dimensional 1H-1H and heteronuclear 31P-1H correlated spectroscopy. The downfield 31P resonance previously noted by Patel et al. (1982) has been assigned by both 17O labeling of the phosphate as well as a pure absorption phase constant-time heteronuclear 31P-1H correlated spectrum and has been associated with the phosphate on the 3' side of the extrahelical adenosine. JH3'-P coupling constants for each of the phosphates of the tridecamer were obtained from the 1H-31P J resolved selective proton-flip 2D spectrum. By use of a modified Karplus relationship the C4-C3'-O3-P torsional angles (epsilon) were obtained. There exists a good linear correlation between 31P chemical shifts and the epsilon torsional angle. The 31P chemical shifts and epsilon torsional angles follow the general observation that the more internal the phosphate is located within the oligonucleotide sequence, the more upfield the 31P resonance occurs. Because the extrahelical adenosine significantly distorts the deoxyribose phosphate backbone conformation even several bases distant from the extrahelical adenosine, 31P chemical shifts show complex site- and sequence-specific variations. Modeling and NOESY distance-restrained energy minimization and restrained molecular dynamics suggest that the extrahelical adenosine stacks into the duplex. However, a minor conformation is also observed in the 1H NMR, which could be associated with a structure in which the extrahelical adenosine loops out into solution. PMID- 2557908 TI - Aspartokinase-homoserine dehydrogenase I from Escherichia coli: pH and chemical modification studies of the kinase activity. AB - The pH variation of the kinetic parameters was examined for the kinase activity of the bifunctional enzyme aspartokinase--homoserine dehydrogenase I isolated from Escherichia coli. The V/K profile for L-aspartic acid indicates the loss of activity upon protonation of a cationic acid type group with a pK value near neutrality. Incubation of the enzyme with diethyl pyrocarbonate at pH 6.0 results in a loss of enzymic activity. The reversal of this reaction by neutral hydroxylamine, the appearance of a peak at 242 nm for the inactivated enzyme, and the observation of a pK value of 7.0 obtained from variation of the inactivation rate with pH all suggest that enzyme inactivation occurs by modification of histidine residues. The substrate L-aspartic acid protects one residue against inactivation, which implies that this histidine may participate in substrate binding or catalysis. Activity loss was also observed at high pH due to the ionization of a neutral acid group with a pK value of 9.8. The reactions of AK HSD I with N-acetylimidazole and tetranitromethane have been investigated to obtain information about the functional role of tyrosyl residues in the enzyme. The acylation of tyrosines leads to inactivation of the enzyme, which can then be fully reversed by treatment with hydroxylamine. Incubation of the enzyme with tetranitromethane at pH 9.5 also leads to rapid inactivation, and the substrates of the kinase reaction provide substantial protection against inactivation. However, three tyrosines are protected by substrates, implying a structural role for these amino acids. PMID- 2557909 TI - Comparison of cooperative and isolated site binding of T4 gene 32 protein to ssDNA by 1H NMR. AB - Deuteriation of all aromatic protons of gene 32 protein (g32P) from phage T4, followed by selective introduction of specific protons, has allowed the precise identification of the number and magnitude of the chemical shift changes induced in the aromatic protons when g32P binds noncooperatively or cooperatively to nucleotides. Signals from five Tyr residues are shifted by binding of g32P to d(pA)8 or d(pA)40-60; however, the change from noncooperative, d(pA)8, to cooperative, d(pA)40-60, binding causes significant increases in the magnitudes of the shifts for only two of these Tyr signals. These two Tyr residues may interact directly with the nucleotide bases, while the shifts associated with the other three Tyr may be due to conformational changes in g32P upon ssDNA binding. Similar conclusions can be drawn for two of the six Phe residues whose protons undergo shifts upon nucleotide binding. Observation of selected proton signals allows for the first time detection by 1H NMR of changes in the proton signals from two Trp residues upon nucleotide binding. The side chains of two Tyr, one or two Phe, and one Trp are probably directly involved in nucleotide base-protein interactions. As assayed by the signals from the H2 and H8 protons of adenine, the bases of a bound nucleotide are undergoing a fast chemical exchange in the noncooperative mode of binding, but shift to slow exchange upon assuming the cooperative mode of ssDNA interaction. When bound to a polynucleotide, the A domain of g32P (residues 254-301) becomes more mobile, as reflected in sharpening of the 1H NMR signals from the A domain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557910 TI - Platelet-neutrophil-smooth muscle cell interactions: lipoxygenase-derived mono- and dihydroxy acids activate cholesteryl ester hydrolysis by the cyclic AMP dependent protein kinase cascade. AB - Fluid-phase interactions between hematologic cells and those of the vessel wall were studied in order to define a role for lipoxygenase products as cell signals in the control of vascular cholesterol metabolism. A functional parameter for hydroxy acids in this system has not been previously demonstrated. We report herein for the first time a biochemical effect of lipoxygenase-derived eicosanoids in the modulation of cholesterol metabolism in smooth muscle cells. Products of platelet-neutrophil interactions served as cell signals in vitro to modulate cholesterol metabolism. We demonstrate that 12-HETE, 12,20-DiHETE, and 12-HETE-1,20-dioic acid activate both lysosomal and cytoplasmic cholesteryl ester (CE) hydrolytic activities, although no effect was observed on CE synthetic (ACAT) activity. The platelet lipoxygenase product, 12-HETE, was the most effective stimulator of CE hydrolysis in the smooth muscle cell, and its conversion to 12,20-DiHETE and the dioic acid derivative by the neutrophils was not necessary for the activation of CE hydrolase. A 2-fold enhancement on CE hydrolysis occurred and was independent of any "cross-activation" by hydroxy acids on production of cyclooxygenase or other lipoxygenase products. The activation of cytoplasmic CE hydrolysis had a lesser cofactor dependence on bile salts in the presence of 12-HETE. This suggested a reduced requirement for surface-active agents in an enzyme-substrate interaction where enzymes are hydrolyzing insoluble lipid substrates. Moreover, 12-HETE induced an additive effect with several lipolytic hormones in the activation of CE catabolism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557911 TI - Role of the stereochemistry of the hydroxyl group of cholesterol and the formation of nonbilayer structures in phosphatidylethanolamines. AB - The phase behavior of mixtures of cholesterol or epicholesterol with phosphatidylethanolamine was studied by differential scanning calorimetry and by X-ray diffraction. Discrete domains of cholesterol are detected by X-ray diffraction in the L alpha phase of phosphatidylethanolamine from egg yolk and synthetic dielaidoylphosphatidylethanolamine beginning at mole fractions of 0.35 0.4 cholesterol. Separate domains of crystalline epicholesterol can also be detected in the L alpha phase of dielaidoylphosphatidylethanolamine by X-ray diffraction at as little as 0.16 mole fraction of epicholesterol. This is a result of poor miscibility of the epicholesterol with dielaidoylphosphatidylethanolamine. Epicholesterol does not alter the L beta----L alpha transition or bilayer spacing. Epicholesterol also has little effect on the diameter of the cylinders in the hexagonal phase. Formation of the inverted hexagonal phase is facilitated by addition of small amounts of cholesterol (mole fraction less than 0.2) in both egg phosphatidylethanolamine and dielaidoylphosphatidylethanolamine. However, at higher mole fractions of cholesterol, the stability of the liquid-crystalline phase is found to increase markedly for dielaidoylphosphatidylethanolamine but not for egg phosphatidylethanolamine, indicating the importance of the structure of the acyl chains in controlling the relative stability of the lamellar and nonlamellar phases in these systems. In contrast to cholesterol, epicholesterol markedly lowers the L alpha----HII phase transition temperature at low mole fraction of sterol. This result demonstrates the importance of the orientation and motional properties of an additive in determining the L alpha----HII transition temperature. PMID- 2557912 TI - Deletion analysis of the mouse m1 muscarinic acetylcholine receptor: effects on phosphoinositide metabolism and down-regulation. AB - Deletions have been constructed in the putative third cytoplasmic loop of the mouse m1 muscarinic acetylcholine receptor (mAChR) gene, and the effects of these mutations on mAChR coupling to phosphoinositide metabolism and agonist-induced down-regulation have been examined following expression in Y1 adrenal carcinoma cells. Deletion of up to 123 of the 156 amino acids in this loop has no effect on antagonist or agonist binding, or on coupling to stimulation of phosphoinositide metabolism. These results suggest that the membrane proximal portions of this loop are involved in determining the specificity of functional coupling of the receptor. Deletion of 75% of the loop has no effect on short-term agonist-induced internalization but does cause a significant decrease in the magnitude of agonist induced down-regulation of receptor number. Thus, this portion of the receptor may be involved in mediating the response to long-term agonist exposure. PMID- 2557913 TI - EPR signals from modified charge accumulation states of the oxygen evolving enzyme in Ca2+-deficient photosystem II. AB - Photosystem II enriched membranes were depleted of Ca2+ and the 17- and 23-kDa polypeptides by treatment with NaCl and EGTA. The 17- and 23-kDa polypeptides were then reconstituted. This preparation was incapable of O2 evolution until Ca2+ was added. An EPR study revealed the presence of two new EPR signals. One of these is a modified S2 multiline signal with an isotropic g value of 1.96 with at least 26 hyperfine peaks (average spacing 55 G) distributed over approximately 1600 G. The other is a near-Gaussian signal with an isotropic g value of 2.004, which is attributed to a formal S3 state. Experiments involving the interconversion of these signals and the effect of Ca2+ and Sr2+ rebinding provide evidence for these assignments. From these results the following conclusions are drawn: (1) These results are consistent with our earlier demonstration that charge accumulation is blocked after formation of S3 when Ca2+ is deficient. (2) Binding of the 17- and 23-kDa polypeptides to photosystem II in the absence of Ca2+ results in the perturbation of the Mn cluster. This is taken as a further indication that the Ca2+-binding site is close to or even an integral part of the Mn cluster. (3) The S3 signal may arise from an organic free radical interacting magnetically with the Mn cluster. However, other possible origins for this signal, including the Mn cluster itself, must also be considered. PMID- 2557914 TI - Specificity of the interaction of amino- and carboxy-terminal fragments of the mitochondrial precursor protein apocytochrome c with negatively charged phospholipids. A spin-label electron spin resonance study. AB - The contribution of the various regions of the mitochondrial precursor protein apocytochrome c to the interaction of the protein with phosphatidylserine dispersions has been studied with chemically and enzymatically prepared fragments of horse heart apocytochrome c and phospholipids spin-labeled at different positions of the sn-2 chain. Three amino-terminal heme-less peptides, two heme containing amino-terminal fragments, one central fragment, and three carboxy terminal fragments were studied. The electron spin resonance spectra of phospholipids spin-labeled at the C5 position of the fatty acid chain indicate that both amino-terminal and carboxy-terminal fragments of the apocytochrome c molecule cause a restriction of motion of the lipids, whereas the heme-containing peptides and protein have less effect. In addition, a second motionally more restricted lipid component, which is observed for apocytochrome c interacting with phosphatidylserine dispersions containing lipids spin-labeled at the C12 or C14 position [Gorrissen, H., Marsh, D., Rietveld, A., & de Kruijff, B. (1986) Biochemistry 25, 2904-2910], was observed both on binding the carboxy-terminal fragments and on binding of the amino-terminal fragments of the precursor protein. Interestingly, even a small water-soluble peptide consisting of the 24 carboxy-terminal residues gave rise to a two-component spectrum, with an outer hyperfine splitting of the restricted lipid component of 59 G, indicating a considerable restriction of the chain motion. This suggests that both the carboxy and amino-terminal parts of the protein penetrate into the center of the bilayer and cause a strong perturbation of the fatty acyl chain motion. The implications of these findings for the mechanism of apocytochrome c translocation across membranes are discussed. PMID- 2557915 TI - Mechanism of adenylate kinase. Is there a relationship between local substrate dynamics, local binding energy, and the catalytic mechanism? AB - Adenylyl (beta,gamma-methylene)diphosphonic acid (AMPPCP) labeled with deuterium at the adenine ring ([8-2H]AMPPCP) and at the beta,gamma-methylene group (AMPPCD2P), as well as adenosine 5'-monophosphate labeled at the adenine ring ([8 2H]AMP), was synthesized and used for deuterium nuclear magnetic resonance (NMR) determination of effective correlation times (tau c) of the free nucleotide and the complexes with adenylate kinase (AK). Extensive and rigorous control experiments and theoretical analysis were performed to justify the validity of the experimental approaches, particularly the fast exchange condition, and the reliability of the tau c values obtained. For the free nucleotide, the results suggest that the phosphonate group of free AMPPCP possesses appreciable local mobility relative to the adenine ring and that complexation with Mg2+ greatly reduced such a local mobility. For the complexes with AK, effective tau c values of 7, 15, 28, 28, and 27 ns were obtained for AMPPCD2P, MgAMPPCD2P, [8-2H]AMPPCP, Mg[8-2H]AMPPCP, and [8-2H]AMP, respectively. These results suggest that the adenine ring of substrates is rigidly bound in all cases, that the phosphonate chain of AMPPCP possesses considerable local mobility, and that Mg2+ reduces such local mobility but does not totally immobilize it. The local dynamics of the analogues bound to AK was correlated with local binding energies for the binding of MgAMPPCP and MgATP to AK estimated from the binding studies by proton NMR and other techniques, in conjunction with the binding theory of Jencks [Jencks, W. P. (1981) Proc. Natl. Acad. Sci. U.S.A. 78, 4046-4050]. The results suggest that no general correlation exists between the local rigidity of portions of a bound substrate and the corresponding (ground state) local binding energy contributed by these portions. In particular, the adenosine moiety contributes little to the binding energy despite the fact that the adenine ring is rigidly bound; the triphosphate (PPPi) moiety behaves oppositely; Mg2+ immobilizes the triphosphate chain but does not enhance binding. Finally, isomers of the substitution-inert beta,gamma-bidentate Cr(III) complexes of adenosine 5'-triphosphate (CrATP) were used to probe two unresolved catalytic problems implicitly related to the local mobility of the phosphonate chain of AMPPCP in the AK-MgAMPPCP complex. The first problem concerns the result of electron paramagnetic resonance (EPR) studies that (Rp)- but not (Sp)-[beta-17O]ATP caused a line broadening in the Mn(II) EPR spectrum of the AK-MnATP complex [Kalbitzer, H. R., Marquetant, R., Connolly, B. A., & Goody, R. S. (1983) Eur. J. Biochem. 133, 221-227].(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2557916 TI - Human deoxycytidine kinase: kinetic mechanism and end product regulation. AB - The kinetic properties of the monomeric deoxycytidine kinase (EC 2.7.1.74) from leukemic human T-lymphoblasts have been investigated. The results of steady-state initial-rate kinetic analysis and product inhibition studies at pH 7.5 and 37 degrees C indicate that substrate binding follows an ordered sequential pathway, with the magnesium salt of ATP being the first substrate to bind and dCMP the last product to dissociate. At subsaturating substrate concentrations, dCMP produced competitive inhibition against ATP, while against varied deoxycytidine concentrations dCMP exhibited mixed-type inhibition. ADP produced noncompetitive inhibition against either substrate. The limiting Km values for deoxycytidine and MgATP were 0.94 and 30 microM, respectively. The end product inhibitor dCTP exhibited competitive inhibition against varied ATP concentration, with a dissociation constant estimated to be 0.7 microM when extrapolated to zero ATP concentration. dCTP was purely noncompetitive against varied deoxycytidine concentration. On the basis of these kinetic results, and on the strong and specific inhibition by dCTP, it is proposed that this end product functions as a multisubstrate analogue, with its triphosphate group binding to the phosphate donor site of the enzyme and its deoxycytidine moiety overlapping and binding to the deoxynucleoside site in a highly specific manner. PMID- 2557917 TI - Stimulation of the topoisomerase II induced DNA cleavage sites in the c-myc protooncogene by antitumor drugs is associated with gene expression. AB - The antitumor drugs mAMSA and VM26 were shown to stimulate the topoisomerase II (Topo II) cleavage activity on the c-myc protooncogene in several human tumor cell lines (N417, HL60, EJ, H146, CaSki, A431, IGROV1, and CAL18A) and human peripheral lymphocytes. The mAMSA-induced gene cleavage was found to increase with the steady-state levels of c-myc transcripts in cell lines while no cleavage could be evidenced in the other genes so far tested. In mAMSA-treated N417 cells, the overall genomic DNA cleavage detected by alkaline elution was found to be about 20 times lower than the c-myc gene cleavage. Topo II mRNA levels were associated with the nuclear Topo II decatenating activity in cell lines and increased with c-myc cleavage. Topo II decatenating activity was found to be 3 times lower in quiescent than in exponentially growing N417 cells, but the c-myc cleavage induced by mAMSA was found as intense in quiescent as in growing cells. Thus, our data seem to indicate that c-myc gene cleavage is not related to cellular Topo II content but rather to c-myc gene transcription. Therefore, we suggest that only a small fraction of the Topo II is able to react with drug on the c-myc gene in relation to its transcriptional accessibility. Since c-myc overexpression is frequently found to be related to human cancer progression, we suggest that this gene could be an important target for Topo II related antitumor drugs. PMID- 2557919 TI - Fe2+ binding to apo and holo mammalian ferritin. AB - The binding of Fe2+ to both apo and holo mammalian ferritin has been investigated under anaerobic conditions as a function of pH. In the pH range 6.0-7.5, 8.0 +/- 0.5 Fe2+ ions bind to each apoferritin molecule, but above pH 7.5, a pH-dependent Fe2+ binding profile is observed with up to 80 Fe2+ ions binding at pH 10.0. This Fe2+ binding is reversible and is accompanied by up to two H+ being released per Fe2+ bound at pH 10.0. The Fe2+ binding to apoferritin probably occurs in the 3 fold channels. A much larger and more complex pH-dependent Fe2+ binding stoichiometry was observed for holoferritin with up to 300 Fe2+ ions binding at pH 10.0. This pH-dependent Fe2+ binding was interpreted as Fe2+ interaction at the FeOOH mineral surface with displacement of H+ from -OH or phosphate surface groups by the incoming Fe2+ ions. Mossbauer spectroscopic measurements using 57Fe labeled Fe2+ under anaerobic conditions showed that 57Fe2+ binding to holoferritin was accompanied by electron transfer to the core, yielding 57Fe3+, presumably bound to the mineral surface. Removal of added iron by Fe2+-specific chelating agents yielded 57Fe2+, demonstrating the reversibility of this electron transfer process. The Fe2+ bound to apo- and holoferritin is readily converted to Fe3+ by exposure to O2 and strongly retained by the respective ferritin species. PMID- 2557918 TI - General mechanism for the bacterial toxicity of hypochlorous acid: abolition of ATP production. AB - The adenylate energy charges (EC) of Escherichia coli 25922, Pseudomonas aeruginosa 27853, and Streptococcus lactis 7962 rapidly fell in nutrient-rich media from values in excess of 0.9 to below 0.1 when the organisms were exposed to lethal levels of HOCl. The same cells maintained in energy-depleted states were incapable of attaining normal EC values necessary for biosynthesis and growth when challenged with nutrient energy sources after HOCl exposure. These changes correlated quantitatively with loss of replicative capabilities. Initial rates of transport of glucose, succinate, and various amino acids that act as respiratory substrates and the ATP hydrolase activity of the F1 complex from the ATP synthase of E. coli 25922 also declined in parallel with or preceded loss of viability. These results establish that cellular death is accompanied by complete disruption of bacterial ATP production by both oxidative and fermentative pathways as a consequence of inhibition of inner membrane bound systems responsible for these processes. PMID- 2557920 TI - Development of nitroxides for selective localization inside cells. AB - The use of nitroxides to measure intracellular phenomena, especially oxygen concentrations, is a new and potentially important approach to a number of physiological and pathophysiological studies. This study provides data indicating the feasibility of developing nitroxides that localize selectively in the intracellular compartment; it is based on the use of readily hydrolysed ester linkages, such that the nitroxides become converted intracellularly to ionic derivatives that do not cross cell membranes readily. Up to 120-fold increased concentrations of intracellular nitroxides (and their one electron reduction product, the hydroxylamines) were obtained. The ESR spectra of the intracellular nitroxides were consistent with their conversion to the ionic species. Preliminary studies indicate that these nitroxides have the properties needed for their use as probes of intracellular concentrations of oxygen and that it should be feasible to synthesize nitroxides that will be even more effective for this purpose. PMID- 2557921 TI - Mechanical and metabolic toxicity of 3-(trimethylsilyl)propanesulfonic acid to porcine carotid arteries. AB - 3-(Trimethylsilyl)propanesulfonic acid (TMSPS) is used as a water-soluble NMR frequency marker. It has its major resonance at 0.00 ppm relative to trimethylsilane, and smaller resonances at 0.62, 1.77 and 2.85 ppm. Its toxicity was tested by exposing contracted porcine carotid strips to increasing concentrations of TMSPS. Up to 3 mM, no statistical change in tension was found. Tension decreased 94 +/- 2% (S.E.) after 30 min in 10 mM TMSPS. An intermediate concentration of TMSPS (6 mM) caused a small fall in phosphocreatine in unstimulated perfused porcine carotid arteries (82 +/- 2% S.E.). A larger decrease (59 +/- 6% S.E.) occurred during K+ contractures in the presence of 6 mM TMSPS. From those experiments it appears the TMSPS is non-toxic in concentrations up to 3 mM, but at greater concentrations inhibits both contraction and phosphorus metabolism. PMID- 2557922 TI - Modulation of cardiac Na channels by angiotensin II. AB - The modulation of Na channels by the vasoactive peptide angiotensin II (AT II) has been studied in isolated ventricular cells of guinea pigs using the patch clamp technique. In cell-attached patches the maximal probability of the channel being open was increased in a concentration range between 0.05 and 1 microM, but decreased at higher concentrations. A maximal increased of 2.5 +/- 0.86 was found at 1 microM AT II. The increase in the probability of the channel being open was due to a decrease in the number of nulls. In all affected cells (n = 17) we observed a delayed inactivation after application of AT II at concentrations between 0.05 and 10 microM. At -30 mV, the time constant of inactivation increased from 1.1 +/- 0.1 ms (controls) to 5.6 +/- 1.6 ms (10 microM AT II). This effect was due to an increased number of openings per sweeps. No significant effect on the mean open time and the first latency were observed. However, due to pronounced bursting, the averaged closed time was significantly increased from 0.8 +/- 0.1 ms to 1.3 +/- 0.1 ms in the presence of 1 microM AT II at -30 mV. An effect of AT II on cardiac Na channels via protein kinase C is discussed. PMID- 2557923 TI - Regulation of the pyruvate dehydrogenase complex by Ca2+ within toluene permeabilized heart mitochondria. AB - (1) Rat heart mitochondria, permeabilized to all low Mr solutes by toluene treatment, have been used to study the regulation in situ of the phosphatase and kinase components of the pyruvate dehydrogenase complex (PDH) by Ca2+. (2) Inactivation of the complex, resulting from phosphorylation by the kinase, and reactivation induced by the phosphatase, were both apparent first-order processes. This behaviour of the phosphatase differs from that observed with toluene-permeabilized adipose tissue mitochondria (Midgley, P.J.W., Rutter, G.A. and Denton, R.M. (1987) Biochem. J. 241, 271-377) where a 'lag phase' preceded reactivation of inactive complex. Further, reactivation due to phosphatase activity was stimulated by Ca2+ only at subsaturating Mg2+ concentrations, in contrast with the extracted enzyme which is stimulated by Ca2+ at all Mg2+ concentrations. (3) Maximum values of half-times observed for inactivation and reactivation were about 10 and 15 s, respectively, at 30 degrees C. (4) At Mg2+ concentrations where effects of Ca2+ on the activity of the phosphatase were apparent, no effect of Ca2+ on the activity of the kinase could be detected. (5) The sensitivity of the phosphatase to [Ca2+] was essentially unchanged in the presence of either ADP or ATP, with half-maximal effects at 0.7 microM in each case. PMID- 2557924 TI - The phosphorylation of p68, a calcium-binding protein associated with the human syncytiotrophoblast submembranous cytoskeleton, is modulated by growth factors, activators of protein kinase C and cyclic AMP. AB - The phosphorylation of the lipocortin-related protein, p68, found in Ca2+ dependent association with the submembranous cytoskeleton has been studied using isolated human placental syncytiotrophoblast plasma membrane vesicles. p68 undergoes rapid, cation-independent phosphorylation in unstimulated membrane vesicles which was inhibited, in a dose-dependent manner, by insulin, platelet derived growth factor, macrophage colony stimulating factor, protein kinase C activating phorbol esters and phosphatidylinositol-specific phospholipase C. Epidermal growth factor had no effect on overall p68 phosphorylation. Transferrin induced an increase in p68 phosphorylation. However, phosphotyrosine was detected in p68 after treatment with epidermal growth factor, macrophage colony stimulating factor or transferrin, whereas a reduction in p68 phosphorylation appeared to be restricted to serine. cAMP and both cholera and pertussis toxins inhibited p68 phosphorylation. Both toxins were synergistic with the effects of insulin and platelet-derived growth factor whilst being antagonistic to the effect of transferrin. Epidermal growth factor and both human and equine immunoglobulin G, all of which alone did not affect overall p68 phosphorylation, reduced cholera or pertussis toxin-induced inhibition of p68 phosphorylation. Several phosphatase inhibitors failed to prevent macrophage colony stimulating factor-induced reduction of p68 phosphorylation. These results indicate that (i) p68 is a potential substrate of receptor tyrosyl kinases, (ii) p68 is not phosphorylated by protein kinase C or cAMP-dependent kinase and (iii) p68 phosphorylation is inhibited by activation of multiple pathways including those employing diacylglycerol or cAMP as second messengers. PMID- 2557925 TI - Gastrin and CCK-8 induce inositol 1,4,5-trisphosphate formation in rabbit gastric parietal cells. AB - The role of phosphoinositide turnover in the mediation of acid secretion was examined in an enriched preparation of isolated rabbit parietal cells (75%). Both gastrin and CCK-8 (octapeptide of cholecystokinin) stimulated [14C]aminopyrine (AP) uptake by cells (EC50 0.07 +/- 0.03 nM (gastrin) and 0.093 +/- 0.065 nM (CCK 8] and increased [3H]inositol phosphates cellular contents (EC50 0.142 +/- 0.016 nM (gastrin) and 0.116 +/- 0.027 nM (CCK-8] in a parallel fashion. In addition, the EC50 values for both phenomenon were quite similar to the Kd values obtained from binding experiments. HPLC analysis of the different [3H]inositol phosphates produced under gastrin or CCK-8 stimulation showed a 2-fold increase in [3H]Ins(1,4,5)P3 levels within 5 s with a concomitant increase in [3H]Ins(1,4)P2 content within 15 s. A low but significant rise in [3H]Ins(1,3,4,5)P4 and [3H]Ins(1,3,4)P3 cellular contents was also observed. No difference between gastrin- and CCK-8-induced inositol phosphates production could be shown. We can conclude that gastrin and CCK-8 display an identical profile of action, suggesting that they stimulate the acid secretory function of parietal cells through the same receptor site coupled to the Ins(1,4,5)P3 production. PMID- 2557927 TI - A monoclonal antibody which inhibits H+/K(+)-ATPase activity but not chloride conductance. AB - A mouse monoclonal antibody was raised against hog gastric membranes. This antibody (95-111 mAb) has a very high affinity for the 95 kDalton band of H+/K(+) ATPase-enriched membranes, and does not react with Na+/K(+)-ATPase. The epitope is located on the tubulovesicles and canaliculi of the parietal cells. The 95-111 mAb also inhibits the ATP hydrolytic activity, decreases the steady-state phosphorylation level and inhibits the phosphatase activity of H+/K(+)-ATPase, strongly suggesting that the epitope is on the catalytic subunit of H+/K(+) ATPase. The 95-111 mAb also recognizes rat, rabbit and human gastric H+/K(+) ATPase. This mAb differs from the H+/K(+)-ATPase-inhibiting mAb previously described (Asano et al. (1987) J. Biol. Chem. 262, 13263-13268), in that it does not inhibit the chloride conductance opened by Cu-o-phenanthroline in gastric vesicles. PMID- 2557926 TI - Developmental patterns in rat brain of phosphatidylinositol synthetic enzymes and phosphatidylinositol transfer protein. AB - Phosphatidylinositol synthetic and intermembrane transfer activities were studied in rat in the developing whole brain and isolated cerebellum. Specific activities of CTP:phosphatidate cytidylyltransferase and CDPdiacylglycerol:inositol phosphatidyltransferase were found to have similar developmental patterns. Levels of phosphatidyltransferase seen in fetal animals (whole brain only) and neonatal (whole brain and cerebellum) were maintained through approximately postnatal day 15, peaked at day 28, and then declined to somewhat higher than fetal levels at day 60. Cytidylyltransferase activity varied from the phosphatidylinositol synthesizing enzyme in that specific activity continued to increase up to day 60. Whole brain phosphatidylinositol transfer specific activity showed a sharp peak at postnatal day 9 after which activity was maintained at or above the fetal levels to day 60. Cerebellum phosphatidylinositol transfer specific activity had a similar peak which was delayed 7-10 days compared to the whole brain. Phosphatidylinositol transfer protein was also determined immunologically: whole brain levels increased dramatically from fetal day 16 to 18 and then remained relatively constant, while cerebellum levels (measured from postnatal day 7) displayed a variable profile between days 7 and 28. The developmental pattern of CTP:phosphatidate cytidylyltransferase in rat brain is reported here for the first time. PMID- 2557928 TI - Inhibitory effects of newly synthesized sulfonamide derivatives on calmodulin dependent phosphodiesterase activity and their modulation of the external ATP dependent permeability change in mammalian cultured cells. AB - External ATP causes a passive permeability change in several types of transformed cells and this change is further enhanced by calmodulin antagonists, such as trifluoperazine. However, such drugs also have nonspecific effects on membrane permeability. We have synthesized several new sulfonamide derivatives, which were found to inhibit calmodulin-dependent phosphodiesterase. The drugs also enhanced the ATP-dependent permeability change in CHO-K1 cells, but their effective concentration ranges were wider than those of previously known antagonists, and thus they would be useful for pharmacological use. PMID- 2557929 TI - [Are the receptors of 1,25-dihydroxyvitamin D3 vitamin K dependent?]. AB - Alimentary deficiency of vitamin K in rats causes a decrease in the level of in vivo occupied nuclear 1,25 (OH)2D3 receptors in small intestinal mucosa and an 2 2.5-fold increase in the ability of cytosolic 1,25 (OH)2D3-receptor complexes to bind to heterologous DNA. The 1,25 (OH)2D3 binding by the receptors is thereby unaffected. Preincubation of kidney and intestinal cytosol of rats with the secondary K-avitaminosis induced by vitamin K antagonist with the microsomal vitamin K-dependent gamma-carboxylation system sharply decreases the binding of the 1.25 (OH)2D3-receptor complexes to DNA. In rats treated with the vitamin K antagonist in combination with a low calcium diet, the subsequent maintenance on a high calcium diet does not cause, in contrast with vitamin K-repleted animals, a sharp decrease of the level of the in vivo occupied 1,25 (OH)2D3 receptors. In vitro Ca2+ cations decrease the binding of the 1,25 (OH)2D3-receptor complexes to DNA only in vitamin K-repleted rats (ED50 = 2.5 x 10(-6) M). The existence of a vitamin K-dependent Ca-sensitive mechanism regulating the binding of the 1,25 (OH)2D3 receptor to DNA has been postulated for the first time. PMID- 2557930 TI - [Study of the inhibition of neutral phosphatase from Pseudomonadaceae by derivatives of its substrates]. AB - The inhibition of neutral phosphatase isolated from the bacteria of the Pseudomonadaceae family by various fragments of the enzyme-hydrolyzed R-O-PO3H2 substrates, inorganic orthophosphate (KH2PO4) and its analogs as well as by adenine, adenosine, alcohols, sugars and amino acids, was studied. It was demonstrated that among other compounds tested only the orthophosphoric acid anions (H2PO4-) exhibit the properties of strong associative inhibitors (K1Vi = 4.35.10(-6)M of the enzyme. The pH dependence of the Michaelis constant [pKm0 = f(pH)] and the inhibition constant for phosphatase by potassium orthophosphate [pK1Vi(KH2PO4) = f(pH)] was studied. The presence in the enzyme active center of a carboxylic (pK = 4.3 +/- 0.1) (presumably, glutamine) and an imidazole (pK = 7.15 +/- 0.1) amino acid residues was postulated. The data obtained were compared to those for neutral, alkaline and acid phosphatases. PMID- 2557931 TI - Insolubilization of apatite-collagen composites by UV irradiation. AB - The physico-chemical properties and biocompatibility of insolubilized apatite collagen composites were examined. A carbonate-containing hydroxyapatite with chemical composition and crystallinity similar to that of bone was synthesized at pH 7.4 and 60 degrees C. The apatite powder was mixed with collagen solution, whose antigenicity had been removed by enzymatic treatment and formed into apatite-collagen pellets. After insolubilization by UV irradiation for 4 h, the composites showed remarkably reduced disintegration and showed good biocompatibility when implanted into rat abdomen. PMID- 2557932 TI - Inactivation properties of T-type calcium current in canine cardiac Purkinje cells. AB - The kinetic behavior of T-type Ca2+ current (ICa-T) was studied in canine cardiac Purkinje cells using a single suction-pipette whole-cell voltage clamp method. ICa-T was studied without contamination of conventional L-type Ca2+ current (ICa L). Ca2+, Sr2+, or Ba2+ were used as the charge carrier. During maintained depolarization ICa-T decayed rapidly, and under most conditions the decay showed a voltage-dependent single exponential time course that did not depend on the species of charge carrier. The development of inactivation did not depend on Ca2+, but the time course required more than a single exponential process. Just negative to the threshold voltage for activating ICa-T, inactivation slowly developed and there was a delay in its onset. The time course of recovery from inactivation was dependent on the protocol used to measure it. As the duration of an inactivating voltage step was increased, recovery slowed markedly and there was a delay in its onset. The time course of recovery could be fit as a biexponential. The fast and slow time constants of recovery were relatively constant, however, the relative amplitudes were dependent on the duration of the inactivating voltage step. Recovery was not dependent on Ca2+, and it was slower at a less negative voltage. These results suggest that the T-type Ca2+ channel in cardiac Purkinje cells follows a complex kinetic scheme dependent only on voltage. This behavior can be accounted for by incorporating into a Markovian model several inactivated and closed states. PMID- 2557934 TI - Surgery of the spine in ankylosing spondylitis. Part I. PMID- 2557933 TI - Polymorphism in the assembly of polyomavirus capsid protein VP1. AB - Polyomavirus major capsid protein VP1, purified after expression of the recombinant gene in Escherichia coli, forms stable pentamers in low-ionic strength, neutral, or alkaline solutions. Electron microscopy showed that the pentamers, which correspond to viral capsomeres, can be self-assembled into a variety of polymorphic aggregates by lowering the pH, adding calcium, or raising the ionic strength. Some of the aggregates resembled the 500-A-diameter virus capsid, whereas other considerably larger or smaller capsids were also produced. The particular structures formed on transition to an environment favoring assembly depended on the pathway of the solvent changes as well as on the final conditions. Mass measurements from cryoelectron micrographs and image analysis of negatively stained specimens established that a distinctive 320-A-diameter particle consists of 24 close-packed pentamers arranged with octahedral symmetry. Comparison of this unexpected octahedral assembly with a 12-capsomere icosahedral aggregate and the 72-capsomere icosahedral virus capsid by computer graphics methods indicates that similar connections are made among trimers of pentamers in these shells of different size. The polymorphism in the assembly of VP1 pentamers can be related to the switching in bonding specificity required to build the virus capsid. PMID- 2557935 TI - The biomechanical evaluation of a new fixation technique for spondylolysis using single and double tension-band wiring. AB - Patients with spondylolysis or spondylolisthesis with persistent symptoms are often marked for surgical treatment. This paper presents the results of a biomechanical study which evaluated the effects of two tension-band wiring methods for this clinical anomaly. Experimental spondylolytic defects were created in canine cadaver lumbar spines, and both wiring techniques were evaluated in cantilever bending. The results demonstrated that experimentally created spondylolytic defects produce a significant decrease in bending stiffness (flexion/extension), and that both the intra- and intersegmental wiring techniques increase the bending stiffness to that of the normal intact spinal segment. PMID- 2557937 TI - Reproducibility of trunk isoinertial performances in the sagittal, coronal, and transverse planes. AB - A new triaxial dynamometer to quantify the performance of trunk muscles has been developed, and a study was conducted to determine the best control parameters to use with this triaxial isodynamic mode of testing. Nine male subjects were tested at three resistance levels in the sagittal, coronal, and transverse planes. The purpose was to establish the reproducibility of the performance parameters in each plane at all resistance levels, and to identify those parameters which gave the most reliable information for objective assessment of the low back functional state. Measured torque had the highest reproducibility rate, and the most reliable assessments were obtained at the highest resistance level. PMID- 2557936 TI - Biomechanics of the Ilizarov external fixator. AB - The rigidity of the Ilizarov external fixator was evaluated by means of the techniques of Briggs and Chao, and compared to their findings for the Hoffman Vidal external factor. A typical fibular fixator consisting of 8 crossed K wires and 4 rings with supporting struts was affixed to a fiberglass-filled epoxy bone fracture model. The ensemble was tested with an MTS servohydraulic testing machine in axial loading, bending (A-P, L-M), and torsion. Displacement transducers were placed on the frame and the bone to determine relative motion. The Ilizarov frame was relatively stiff in compression; failure occurred at about 100 kg due to slippage at the wire holders. In bending, it was much less rigid than the Hoffman-Vidal fixator due to bowing of the transverse wires and slippage of the bone along these wires. Stiffness is related to the wire-bone orientation: wires parallel or nearly parallel to the applied force provide little resistance to deformation. In torsion, the laxity in the system is due primarily to wire deflection or wide spacing between adjacent rings. Stiffness in compression and bending increased as a function of wire tension to about 130 kg (further tightness was not possible due to slippage at the wire holder). The Ilizarov fixator is less rigid than other fixators in all loading modes, particularly in axial compression. This may prove to be clinically beneficial as evidenced by increased osteosynthesis. However, the existing system has many sites of laxity. Care must be taken in frame construction to ensure adequate stability and necessary stiffness. PMID- 2557938 TI - The balance point of the intervertebral motion segment: an experimental study. AB - A loading or "balance" point was sought that could serve as a functional reference for mechanically testing spinal motion segments. This point is located above the in-vitro motion segment where, when an axial compressive load is applied, the segment exhibits minimal coupled rotation. The balance point is a reliable indicator of the mechanical characteristics of the segment. Segments exhibited increasing rotation as axial compressive loads were applied further and further away from the balance point. The location of the balance point was significantly affected by sustained static or cyclic flexion-compression loading and by brief flexion-compression overloads. PMID- 2557939 TI - Zirconia: the second generation of ceramics for total hip replacement. AB - Alumina ceramic was successfully introduced into orthopaedic surgery for total hip replacement 20 years ago. Its mechanical properties are currently optimized, and there is a need to develop new ceramics having higher mechanical properties, mainly in tension. Yttrium-oxide partially stabilized zirconia is one of the new ceramics that exhibits much more toughness than alumina. This article reviews the processing as well as the mechanical and biological properties of this new class of materials which are already in clinical use in Europe. PMID- 2557940 TI - Hydroxyapatite ceramics and the nature of the bone-ceramic interface. AB - Hydroxyapatite ceramics have been used as bone implants in a number of experimental systems and clinical applications. We have developed a unique experimental model that allows study of the interface between bone and implant materials. A comparison of titanium and hydroxyapatite materials, using this model, has demonstrated the osteoconductive nature of hydroxyapatite and its ability to bond directly to bone. PMID- 2557942 TI - A resolution restriction for Wolff's law of trabecular architecture. AB - A rigid constructionist view of Wolff's trajectorial theory of trabecular architecture, called the "orthodox theory," requires that the trabeculae of cancellous bone intersect at right angles exactly like the stress trajectories with which they are associated by the theory. It is well established that trabeculae do not always intersect at right angles. The author discusses the fallacy in the orthodox interpretation of Wolff's law, and suggests that a "resolution length restriction" be imposed on the trajectorial theory to avoid interpretations that lead to the fallacy. PMID- 2557941 TI - The effect of aluminum and gallium ions on the mineralization process. AB - Metal ions have various and significant effects on the skeletal system. Aluminum accumulation in renal dialysis patients causes osteomalacia, while gallium is an effective therapeutic agent for treating the hypercalcemia accompanying certain malignancies. Using in-vitro systems that stimulate in-vivo mineralization, the authors have investigated the physical-chemical mechanisms of the actions of aluminum and gallium and report some of their findings. PMID- 2557943 TI - The use of adhesives in chondrocyte transplantation surgery: in-vivo studies. AB - Two commercial adhesive preparations--fibrin glue and mussel adhesive protein (MAP)--were tested in-vivo for their ability to fix a chondrocyte allograft internally. While results for the fibrin, including additional testing for chondro inductive/conductive properties, were at best inconclusive, the results for MAP are highly promising. PMID- 2557944 TI - Rape and subsequent seroconversion to HIV. PMID- 2557945 TI - Pathway and kinetics of vitellogenin-gold internalization in the Xenopus oocyte. AB - After in vitro incubation of Xenopus oocytes with vitellogenin (VTG)-gold conjugate, the gold particles are distributed on the whole plasma membrane. Their concentration in coated pits still occurs at 0 degrees C. At +20 degrees C the label quickly (30 sec) appears in multi-vesicular endosomes (MVE) which segregate together with primary endocytic vesicles into distinct clusters below the plasma membrane. From this step up to crystallization of the yolk platelets, the gold particles stay in the same compartment. During 5.5 h the label progressively increases along the MVE membrane, first (1.5 h) by fusion of primary endocytic vesicles with consecutively enlarging endosomes, then (4 h) by decreasing of the MVE membrane. As concerns the yolk platelet formation, concentration of primordial yolk platelets (PYP) occurs at 5.5 h from the incubation onset, the labeling of preexisting yolk platelets starts at 7 h, while crystallization of PYP begins only after 12-13 h. Our results indicate that VTG receptors are not preclustered in coated pits and their lateral translation is not inhibited at 0 degrees C. The yolk protein processing takes place within one compartment only. The VTG condensation begins with a long concentration phase of receptor-VTG complexes still integrated in the endosome membrane. It occurs in MVE by: i) a repeated fusion of primary endocytic vesicles; ii) removing part of the endosome membrane by internal vesiculation. Fusion between endosomes occurs only after VTG has dissociated from its receptors and VTG dissociates only when when the density of the VTG-receptor complexes in the endosome membrane is sufficient. Crystallization begins after a 7-8 h delay. The endosome migration into the oocyte is also controlled by the binding of VTG to its receptors. Our results also demonstrate that binding of VTG colloidal gold modifies neither the vitellogenic pathway nor the duration of the vitellogenin internalization. However when vitellogenin is bound to colloidal gold, dissociation of ligand receptor complexes is delayed because the amount of ligand in the incubation medium is necessarily low. PMID- 2557946 TI - Microfilaments and cellular signal transduction: effect of cytochalasin D on the production of cAMP, inositol phosphates, and on calcium movements in rat parotid glands. AB - In rat parotid glands, the involvement of the microfilament system in the cellular signal transmission mechanism was tested by measuring the effect of cytochalasin D (which disturbs the microfilament system) on the production of intracellular second messengers. Cytochalasin D (CD) did not affect unstimulated calcium movements (measured by the 45Ca efflux technique) or inositol phosphate production or cAMP accumulation. Neither did it modify the generation of intracellular second messengers induced by activation of the cholinergic muscarinic receptor (calcium and inositol phosphates). CD dit not affect the cAMP accumulation induced by the activation of the beta-adrenergic receptor whereas it strongly inhibited the calcium movements induced by activation of the same receptor. These data suggest that, in rat parotid glands, calcium movements, induced by beta-adrenergic receptor stimulation need an intact microfilament system to occur, whereas the muscarinic pathway (via IP3) does not. PMID- 2557947 TI - Evaluation of an alpha agonist alone and in combination with a nonsteroidal antiinflammatory agent in the treatment of experimental rhinovirus colds. AB - The pathogenesis of symptoms of the common cold and their optimal treatment are incompletely understood. To evaluate the role of an oral alpha agonist alone and in combination with a nonsteroidal anti-inflammatory drug in the treatment of experimental rhinovirus colds, 58 subjects were randomized to receive pseudoephedrine 60 mg alone, pseudoephedrine 60 mg plus ibuprofen 200 mg, or placebo, four times daily for 4 1/2 days beginning 30 hours after intranasal rhinovirus inoculation under double-blind conditions. The frequencies of infection, colds occurrence, and viral shedding did not differ significantly between the groups. Total symptom scores were reduced by 59% by pseudoephedrine plus ibuprofen (p less than 0.05) and 48% by pseudoephedrine alone compared with placebo. Nasal symptom scores tended to be lower in recipients of pseudoephedrine plus ibuprofen compared with pseudoephedrine alone (p = 0.09), but other parameters showed no significant treatment differences between the groups. Rhinorrhea, as determined by nasal secretion weights, was significantly reduced in both treatment groups compared to placebo. Nasal patency measurements tended to show the greatest improvement in recipients of pseudoephedrine plus ibuprofen. Therapy was clinically well tolerated. The results suggest that an oral alpha agonist is effective in modifying certain manifestations of experimental rhinovirus infection and that the addition of a nonsteroidal anti-inflammatory drug may provide additional benefit in nasal symptoms and patency. Studies involving large numbers of patients with natural colds are needed to determine the clinical significance of these findings. PMID- 2557948 TI - Immune modulation by aspirin during experimental rhinovirus colds. PMID- 2557950 TI - A comparison of scar quality in wounds closed under tension with PGA (Dexon) and Polydioxanone (PDS). AB - Wounds closed under tension have a tendency to stretch. To assess whether this stretching can be limited by inserting absorbable subcuticular sutures, 28 wounds, which involved elliptical excisions, were closed with PGA and PDS sutures in opposite halves of the same wound. PGA scars were found to be significantly wider than their PDS counterparts and were associated with greater hypertrophy at 6 months. PMID- 2557949 TI - Calcium homeostasis and the activation of calcium channels in cells of the immune system. AB - In summary, calcium seems to play a central role in the activation of cells of the immune system. When the cells are stimulated, [Ca2+]i generally increases as a result of entry from the external medium, as well as mobilization of calcium from intracellular membrane-bound compartments. It is generally acknowledged that inositol 1,4,5-trisphosphate mediates the release of calcium from internal stores. By comparison, relatively little is known about the mechanism underlying entry of calcium across the plasma membrane. Currently available evidence seems to rule out the participation of classic voltage-gated channels. Similarly, it appears unlikely that the interaction of ligands with their surface receptors directly activates the channels. Instead, it is more likely that a second messenger produced by the formation of the receptor-ligand complex is responsible for gating the channels. Several mediators, including calcium itself and inositol phosphates, have been proposed to effect gating of the channels, but conclusive evidence is not yet available. Ongoing work is likely to reveal the nature of the putative second messenger(s) in the near future. PMID- 2557951 TI - Periventricular enhancement: a non-pathognomonic sign of intracerebral tumours. AB - Four cases of periventricular contrast enhancement on computed tomography due to different tumours are reported, emphasizing that periventricular contrast enhancement is a non-specific radiological sign. PMID- 2557952 TI - A localized hypersensitivity reaction to co-trimoxazole in a previously irradiated field simulating a recall phenomenon. PMID- 2557953 TI - Medullary carcinoma of the thyroid: management of persistent hypercalcitonaemia utilizing [99mTc] (v) dimercaptosuccinic acid scintigraphy. AB - Whole body scintigraphy with [99mTc] (v)dimercaptosuccinic acid (pentavalent DMSA) was performed in seven patients with histologically confirmed medullary carcinoma of the thyroid (MCT). Six of these patients had undergone previous thyroid resections for MCT and, although asymptomatic at the time of pentavalent DMSA scintigraphy, had persistent and serial elevations in their plasma calcitonin levels. One additional patient was scanned before and after total thyroidectomy for MCT. The pentavalent DMSA scintigram demonstrated either local neck recurrence (three patients) or distant metastases (two patients) in five of the six asymptomatic patients. In one asymptomatic patient only equivocal neck uptake was demonstrated. Since he had only minimal calcitonin elevations, repeat neck exploration was not performed. The one patient studied before thyroid resection for MCT demonstrated neck uptake before, but not after, total thyroidectomy. The results of the scintigrams had significant impact on patient care and resulted in neck re-exploration (three patients), neck biopsy (one patient), and lumbar spine biopsy and subsequent radiotherapy (one patient). These data demonstrate pentavalent DMSA to be a sensitive localizing agent in the evaluation of asymptomatic MCT patients with hypercalcitonaemia. Accurate targeting of treatment may be shown in due course to have a beneficial impact on survival. PMID- 2557954 TI - Calcifectomy for ductal carcinoma in situ. PMID- 2557955 TI - Paget's disease. PMID- 2557956 TI - Characterization of rat spinal cord receptors to FLFQPQRFamide, a mammalian morphine modulating peptide: a binding study. AB - An in vitro binding assay, using 125I-YLFQPQRFamide, a newly synthetized iodinated analog of FLFQPQRFamide, in which Phe1 (F) has been substituted by a Tyr (Y), was developed to demonstrate and characterize putative binding sites of this brain morphine modulating peptide. This radioligand bound in a time dependent manner to rat spinal cord membrane preparation. This binding was dose dependent, saturable and reversible. Both kinetic data and saturation measured at equilibrium lead to the existence of a homogenous population of high affinity binding sites with a Kd value of 0.09-0.1 nM and a maximal capacity Bmax of 14.5 +/- 2 fmol/mg protein. Results of competition experiments show that both FLFQPQRFamide and its analog YLFQPQRFamide had a similar capacity to inhibit the 125I-YLFQPQRFamide binding, suggesting that this radioiodinated analog is a good tool to study binding characteristics of FLFQPQRFamide receptors. The related octadecapeptide AGEGLSSPFWSLAAPQRFamide, another mammalian morphine modulating peptide competes for radioligand binding with similar potency. Our results also show that mu, delta and kappa opiate receptor agonists as well as the antagonist naloxone were not able to affect binding either in presence or in absence of 120 mM NaCl. Together, these data demonstrate that FLFQPQRFamide does not function as an endogenous opiate receptor antagonist and that is capacity to reduce opiate induced analgesia is supported by specific binding sites. PMID- 2557957 TI - Involvement of N-methyl-D-aspartate (NMDA) receptors in respiratory rhythmogenesis. AB - The involvement of N-methyl-D-aspartate (NMDA) subtype of glutamate receptors in the control of inspiratory termination was studied in paralyzed decerebrated cats. Cats were either vagotomized, or had intact vagus nerves and were ventilated with a ventilator driven by the discharge of the phrenic nerve. The systemic administration of NMDA antagonists acting non-competitively (MK-801, ketamine, phencyclidine) or competitively (2-amino-7-phosphonoheptanoic acid: AP7), produced an apneusis in vagotomized animals or in animals transiently deprived of vagal pulmonary feedback by the 'no inflation test'. After NMDA receptor blockade, the inspiratory phase could be terminated by lung inflation or sensory stimulation. Thus pharmacologically distinct mechanisms control the termination of inspiration: vagal afferents which are NMDA-independent, and a central mechanism acting through the activation of NMDA receptors. The apneustic pattern induced by NMDA receptor blockade was characterized by a decrease of the amplitude of integrated phrenic nerve activity, the persistence of CO2 sensitivity and an enhancement of apneusis by anaesthesia. After injection of NMDA antagonists there was a decrease of the duration of expiration which thereafter remained constant and dissociated from inspiratory duration. The possible mechanisms by which NMDA receptors may contribute to respiratory rhythmogenesis are discussed. PMID- 2557958 TI - Canine narcolepsy is associated with an elevated number of alpha 2-receptors in the locus coeruleus. AB - alpha 2-Receptors in the canine brain were pharmacologically characterized using [3H]yohimbine binding. Competition studies revealed a single class of binding sites in frontal cortex but two distinct subtypes in nucleus caudatus. The role of central alpha 2-receptors in narcolepsy was investigated in 5 normal and 5 narcoleptic Doberman pinschers. Scatchard analysis of [3H]yohimbine binding in different brain areas revealed an increase in the number of alpha 2-binding sites limited to the locus coeruleus. This suggests that altered autoinhibition of norepinephrine release may be associated with the narcoleptic symptomatology. PMID- 2557960 TI - Localization of oxytocin binding sites in the human brainstem and upper spinal cord: an autoradiographic study. AB - Two different ligands, tritiated oxytocin and a newly synthesized and monoiodinated oxytocin antagonist, were used to reveal sites which bind oxytocin in the brainstem and upper spinal cord of 12 human subjects. Tissue sections were incubated with either ligand at a concentration close to their respective dissociation constants determined in human uterus and rat brain. Specificity of binding was assessed in presence of unlabelled oxytocin in excess. Comparable results were obtained using tritiated or iodinated ligand. Labelling was most intense in the substantia nigra pars compacta, the substantiae gelatinosae of the caudal spinal trigeminal nucleus and of the dorsal horn of the upper spinal cord, as well as in the medio-dorsal region of the nucleus of the solitary tract. Binding was also detected in the rest of the nucleus of the solitary tract and in other areas, including the oral and interpolar parts of the spinal trigeminal nucleus, the hypoglossal nucleus and the area postrema. Presence of oxytocin binding sites in regions concerned with sensory, autonomic and motor processing suggests that oxytocin could act as a neurotransmitter or neuromodulator in the human central nervous system. PMID- 2557959 TI - Calcium-dependent displacement of haloperidol-sensitive sigma receptor binding in rat hippocampal slices following tissue depolarization. AB - To evaluate the possible existence of an endogenous ligand for the haloperidol sensitive sigma receptor, we developed an in vitro competition assay to measure endogenous ligand release. Depolarization of in vitro hippocampal slices by either veratridine or potassium reduced [3H]ditolylguanidine binding in a calcium dependent and transient manner. None of the drugs or iron substitutions directly affected [3H]ditolylguanidine binding to rat brain membranes. Veratridine-induced depolarization also reduced the binding of [3H](+)3-(3-hydroxyphenyl)-N-(1 propyl)piperidine, another sigma radioligand, in a calcium-dependent manner. Radioligand displacement was not associated with alteration in sigma receptor dissociation kinetics or receptor degradation in the hippocampal slice. In contrast, KC1 depolarization had no effect on [3H]ditolyguanidine binding to sigma receptors in liver slices. The results suggest that a calcium-dependent, depolarization-induced reduction in sigma receptor binding may have been caused by the release of an endogenous sigma ligand in rat hippocampal tissue. PMID- 2557962 TI - The N-methyl-D-aspartate receptor complex in Alzheimer's disease: reduced regulation by glycine but not zinc. AB - The binding of [3H]MK-801 to the N-methyl-D-aspartate receptor complex of well washed cortical membranes from brains of examples of Alzheimer's disease and controls has been determined in incubations containing either glutamate or glycine plus glutamate. No changes were detected in the IC50 values for inhibition by zinc in the Alzheimer's samples compared to control although 'glycine-dependent' binding of the [3H]-ligand was significantly reduced in Alzheimer's disease. PMID- 2557963 TI - Alpha-receptor and cholinergic receptor-linked changes in cytosolic Ca2+ and membrane potential in primary rat astrocytes. AB - Both phenylephrine and carbachol caused a sustained increase in Ca2+ influx and intracellular free Ca2+ of primary astrocytes as measured with 45Ca2+ and fura-2. The responses to phenylephrine and carbachol were additive, suggesting that they use different releasable pools of Ca2+. If extracellular Ca2+ was removed by EGTA only a transient rise in cytosolic Ca2+ was seen upon application of the agonists. Both compounds caused depolarization of the astrocyte membrane as determined with the optical probe 3,3-diethylthiadicarboxyamineiodide. Activation of protein kinase C with 12-tetradecanoylphorbol myristate acetate (TPA) or the diacylglycerol analogue dioctanoylglycerol (DiC8) also depolarized the cells. A prior activation of protein kinase C with TPA or DiC8 abolished the depolarizing effect of phenylephrine suggesting that they act through the same mediators. If the cells were made ideally permeable to K+ with the ionophore valinomycin, or the K+ channels had been blocked with Ba2+, neither TPA nor phenylephrine had any significant effect on the membrane potential. Neither TPA nor phenylephrine had any effect on the 86Rb+ equilibrium potential across the cell membrane. The results suggest that the depolarizing effect of these substances could be through a blocking of K+ channels. PMID- 2557964 TI - Regulation of alpha-melanocyte-stimulating hormone release from superfused slices of rat hypothalamus by serotonin and the interaction of serotonin with the dopaminergic system inhibiting peptide release. AB - Release of alpha-melanocyte-stimulating hormone (alpha-MSH) from frontal slices of rat hypothalamus superfused with oxygenated artificial cerebrospinal fluid (ACSF) was quantified by radioimmunoassay. Control depolarisations with 50 mM KCl containing ACSF produced significant increases in alpha-MSH release which were partially blocked by 10(-6) M cinanserin, a serotonin (5-HT) receptor antagonist. Superfusion of the tissues with varying concentrations of 5-HT (10(-7) M to 10( 4) M) resulted in an inverted U-shaped dose-response curve, maximum alpha-MSH release being obtained with 10(-6) M 5-HT. Addition of 10(-6) M cinanserin shifted the 5-HT dose-response curve to the right whilst the presence of 10(-8) M flupenthixol, a dopamine receptor antagonist, resulted in a sigmoidal 5-HT dose response curve. Superfusion with ACSF containing either 10(-7) M fluoxetine, a 5 HT re-uptake inhibitor, or 10(-7) M p-chloroamphetamine, an agent releasing 5-HT, induced significant increases in alpha-MSH release which were abolished in the presence of 10(-6) M cinanserin. These data demonstrate the presence of an endogenous 5-HT system that exerts a biphasic effect on alpha-MSH release. A stimulatory effect caused by lower 5-HT concentrations appears to be a direct action whilst an inhibitory effect at higher concentrations is mediated through an inhibitory endogenous dopaminergic system. A significant proportion of K+ stimulated peptide release is 5-HT-mediated. PMID- 2557961 TI - Ubiquitin in normal, reactive and neoplastic human astrocytes. AB - Ubiquitin, a protein important in regulating non-lysosomal proteolysis, has previously been shown to be present in cytoskeletal inclusions of the neurodegenerative diseases. Its role in other pathological processes of the central nervous system, such as neoplastic transformation of cells, is not known. The astrocytoma, a tumor of complex biology derived from the astrocyte, is the most common primary parenchymal human brain tumor in both children and adults. Until recently, ubiquitin was not known to form stable conjugates in cells. We have shown using immunocytochemistry on sections of astrocytomas that both glial fibrillary acidic protein (GFAP) (the major intermediate filament protein present in normal, reactive and neoplastic astrocytes) and ubiquitin are simultaneously present in the cytoplasm and cell processes of tumor cells. The presence of ubiquitin and GFAP was also found in astrocytoma cells in short- and long-term culture, and confirmed by immunostaining of blots of tumor homogenates subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis. PMID- 2557965 TI - Autoradiographic mapping of a selective cyclic adenosine monophosphate phosphodiesterase in rat brain with the antidepressant [3H]rolipram. AB - Rolipram is a clinically effective antidepressant with selective cAMP phosphodiesterase (PDE) inhibiting properties. (+/-)-[3H]Rolipram binds with high affinity (Kd = 2.52 +/- 0.47 nM) to sections of rat brain (Hill number = 0.90 +/- 0.05). Binding is stereospecific. Association of (+/-) [3H]rolipram to sections is rapid (47% of specific binding in the first minute, kobs = 0.52 min-1). Dissociation of (+/-)-[3H]rolipram exhibits non first order kinetics (3 component model; t1/2 = 2.5 min, 50 min and 6 h, respectively). A number of PDE inhibitors reduce (+/-)-[3H]rolipram binding to the level of nonspecific binding ((-) rolipram, IC50 = 0.9 nM; (+/-)-rolipram, IC50 = 1.5 nM; Ro 20-1724, IC50 = 11 nM; ICI 63.197, IC50 = 35 nM; medazepam, IC50 = 240 nM; diazepam, IC50 = 1200 nM; IBMX, IC50 = 3800 nM). In vitro autoradiography reveals high binding site densities in the cerebellum, olfactory bulb, lateral septal nucleus, frontal cortex, subiculum and CA1 of hippocampus. Most of the labeled structures are part of the limbic system. In vivo autoradiography of (+/-)-[3H]rolipram binding shows much more nonspecific binding than in vitro, nevertheless the distribution pattern of (+/-)-[3H]rolipram binding sites is similar. A comparison of the distribution pattern of (+/-)-[3H]rolipram binding sites with that of an antidepressant (monoamine oxidase inhibitor, monoamine uptake inhibitor) reveals no overlap. Limited, though significant correlations exist with the distribution of beta 1-adrenergic, adenosine1 and glutamate/quisqualate receptors as well as protein kinase C, but not with beta 2-adrenergic receptors and forskolin binding sites. PMID- 2557966 TI - [35S]TBPS binding sites are decreased in the colliculi of mice with a genetic predisposition to bicuculline-induced seizures. AB - Several differences have been found in GABAergic function between the long sleep (LS) and short sleep (SS) mice which were genetically selected for different ethanol-induced sleeptimes, and it has been suggested that these differences may explain their differential ethanol sensitivity. However, these lines also differ in seizure susceptibility, a behavior which may also be mediated by GABAergic pathways. Thus, it is difficult to associate differences in GABA neurochemistry with either of these behaviors, particularly when only two selected lines are used. We measured differences in the density and affinity of the [35S]TBPS binding site on the GABAA receptor/Cl- ionophore complex in discrete brain areas; and in order to determine the relationship between receptor binding and behavioral differences, we included mice from 5 of the LS and SS recombinant inbred strains (LS x SS RI) in addition to mice from the LS and SS lines. [35S]TBPS binding in sagittal brain sections was analyzed by quantitative autoradiography, and the amount of binding differed depending on whether bicuculline was added to inhibit endogenous GABA binding. In the presence of bicuculline, the number of [35S]TBPS sites in SS mice was highest in the colliculi (4.5 +/- 0.5 pmol/mg protein), cerebellum (4.8 +/- 0.6 pmol/mg), hippocampus (3.2 +/- 0.7 pmol/mg) and cortex (2.9 +/- 0.3 pmol/mg). The Bmax was two-fold lower in both superior and inferior colliculi (IC) of LS mice. There were no differences between lines in Bmax in any other area and in Kd values in any area (58 +/- 4.0 nM).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557968 TI - Specific lesions of rat spinal interneurons induced by systemic administration of acromelic acid, a new potent kainate analogue. AB - A single systemic injection of acromelic acid, a potent kainate analogue, caused behavioral and pathological effects distinct from those seen after systemic kainate. There was an initial marked tonic extension of the rat hindlimb, followed often by convulsions and, in surviving rats, by a transient flaccid paralysis and, ultimately, a persistent spastic paraplegia. Pathological examination suggested specific lesions of interneurons in the lower spinal cord with little or no damage to the hippocampal neurons preferentially affected by systemic kainate. PMID- 2557967 TI - Insulin-like growth factor 1 (IGF-1) receptors in the human brain: quantitative autoradiographic localization. AB - The distribution of Insulin-like growth factor 1 (IGF-1) receptors in large cryosections of human brain hemispheres (80-microns) was studied by quantitative autoradiography using 125I-IGF-1 as ligand. Postmortem tissue only from individuals free from neurological diseases was used. The highest densities of IGF-1 receptors were found in the hippocampus, amygdala and parahippocampal gyrus. Intermediate densities were observed in the cerebellum, cerebral cortex and caudate nucleus, whereas low densities of IGF-1 receptors were obtained in the substantia nigra, red nucleus, white matter and cerebral pedunculus. The cartography of IGF-1 receptors in the normal human brain will hopefully be of use in the study of the alteration of these receptors in diseased brain. PMID- 2557969 TI - [Biochemical characteristics of voltage-dependent sodium channels]. AB - Present knowledge on voltage dependent sodium channel is summarized in review form. Isolation procedures of channel proteins, subunit composition, posttranslational processing, distribution in various tissues, as well as channel subtypes are being dealt with. Further the properties of the native and reconstituted channels are compared and modifications of channel functions with some pharmacological agents are considered. PMID- 2557970 TI - [Role of liposomes in the presentation of HIV envelope glycoprotein and the immune response in mice]. AB - The role of antigen presentation in the induction of humoral as well as cell mediated immune responses has been investigated by anchoring HIV-1 envelope glycoprotein (gp 160/120) into the phospholipidic bilayer of preformed liposomes to produce HIV-Immunosomes. HIV-Immunosomes induced high titres of HIV-specific antibodies when tested by ELISA, IFA and neutralization, whereas equal amounts of purified glycoprotein alone produced lower antibody response. Similarly, HIV Immunosomes induced antigen-specific Interleukin-2 production and blastogenic response upon restimulation with the same antigen, in animals vaccinated with HIV Immunosomes, whereas no secondary response was observed in animals vaccinated with equal amounts of purified gp 160/120. Taken together, these results underline the importance of antigen presentation in the establishment of an adequate immune status and show the potential of HIV-Immunosomes as vaccine against AIDS. PMID- 2557971 TI - [Cloning and sequence analysis of cDNA of common precursors of three hypothalamic neuropeptides immunologically related to human somatocrinin 1-37, alpha melanotropin and salmon melanin-concentrating hormone]. AB - We have cloned and sequenced DNAs complementary to the mRNA encoding the precursor of the rat melanin-concentrating hormone. This allowed us to elucidate the primary structure of the 96 C-terminal residues of this precursor. It contains three possible sites for enzymatic cleavage enabling the generation of MCH and of two additional neuropeptides. These three peptides can bind one of our antisera raised against human somatocrinin 1-37, alpha-melanotropin and salmon melanin-concentrating hormone, which immunocytochemically stain the same neuron population in the hypothalamus. PMID- 2557972 TI - [Antibiotic resistance plasmids from Serratia marcescens and their elimination by DNA-gyrase inhibitors]. AB - We studied the effects of ciprofloxacin on 73 strains of Serratia marcescens. In the first place, we have tested their plasmidic content: 76% of Serratia marcescens strains contained plasmids by electrophoresis, and 29% of these plasmids were self-transmissible by conjugation. Secondly, we studied the plasmid stability with regard to ciprofloxacin. We obtained a spontaneous cure with 19% of plasmids, and ciprofloxacin, at very low concentrations (0.4 mg/l), increased the rate of cure and more efficiently than novobiocin, a compound used as a known curing agent. PMID- 2557973 TI - Staphylococcal alpha toxin induced cardiac dysfunction. AB - Myocardial dysfunction in isolated heart muscle preparations from Wistar rats was induced by low concentrations (3 hemolytic units/mL) of staphylococcal alpha toxin. Exposure of isolated, intact hearts to alpha toxin caused an increase in coronary perfusion pressure within 1 min that continued to rise throughout the experiment. Similar changes occurred with intraventricular systolic pressure. Diastolic pressure initially remained stable but increased after 5 to 6 mins of treatment. Coronary pressure increased approximately 68% while ventricular contractile response was altered 45%. Short exposure to alpha toxin, followed by perfusion with toxin-free buffer did not reverse the coronary vascular response. Pretreatment of the heart with the same concentration of alpha toxoid blocked alpha toxin. Tension development in isolated atria was insensitive to 3 hemolytic units/mL of alpha toxin but it was altered by 12 hemolytic units/mL. Proteinaceous extracts from ventricular cardiac muscle inhibited alpha toxin fourfold more than extracts from atrial tissue. These data demonstrate that purified alpha toxin, administered in low concentrations to cardiac muscle with an intact vascular supply, irreversibly changes pump function primarily through vascular constriction. Direct effects on cardiac muscle tissue could only be observed with substantially higher toxin concentrations. The mechanism of action appears to be mediated through a binding or receptor response that is more sensitive in ventricular than atrial muscle. PMID- 2557974 TI - The role of excitatory amino acids and intraneuronal calcium in the acute intoxicational effects of ethanol. PMID- 2557975 TI - [Influence of chronically-administered enalapril on isotonic and isometric exercise in patients with moderate grade hypertension]. AB - We studied 11 patients affected by mild essential hypertension during chronic therapy with enalapril (E). After a pharmacological wash-out the patients were treated with E once a day (10-20 mg) for 4 weeks. Before and after the treatment period the patients were studied by means of the isotonic exercise stress test on the cycloergometer with increments of 25 W every 2 min and by means of the Sustained Handgrip test (SHG) at 70% of maximal capacity for 1 min. During the study period E reduced the blood pressure at rest in all patients without untoward effects. During the isotonic test and particularly during SHG, E reduced systolic and diastolic blood pressure (BP) and the product systolic BP x heart rate. The treatment did not influence the time length of the isotonic exercise stress test. Our results suggest that E does not increase the MVO2 at rest and during different types of exercise: this can be very important because many patients affected by hypertension suffer from ischemic heart disease. E is utilized also in patients with heart failure, some of whom have an hypertensive or ischemic cardiomyopathy. PMID- 2557977 TI - Indications for use of porous hydroxyapatite at time of endosteal implant placement. AB - Placement of coralline porous hydroxyapatite (PHA) at the time of first stage endosteal implant surgery has been shown to be effective in preventing mucosal dehiscence when the buccal cortex is perforated during implant placement. PHA was used to obliterate the space around the occlusal end of endosteal implants placed during tooth extraction. When vital structures would be jeopardized by full implant insertion, PHA can be placed in the area around the protruding implant to prevent mucoperiosteal flap fenestration. PMID- 2557976 TI - [Evaluation of the cardiac and peripheral vascular effects of the single-dose administration of enalapril and captopril in essential arterial hypertension]. PMID- 2557978 TI - [The effect of colchicine and its derivatives on the activity of enzyme markers of T-lymphocytes]. AB - Colchicine and its less toxic derivatives (demecolcine and deacetyl colchicine), in dependence on the concentration in the incubation medium, inhibited the activity of adenosine deaminase in the lymphocytes. All the three substances increased the level of cyclic adenosine monophosphate which has the ability of influencing the proliferation of T-lymphocytes. Colchicine and its derivates also inhibited the activity of purine nucleooxidophosphorylase, which, similarly as adenosine deaminase, is a marker enzyme of T-lymphocytes. The results obtained lead to the assumption that the mentioned preparations inhibit the function of T lymphocytes. PMID- 2557979 TI - The podosomes of Rous sarcoma virus transformed chondrocytes show a peculiar ultrastructural organization. AB - The ultrastructure of F-actin-containing punctate adhesion structures (podosomes) and of their rosette-like clusters has been studied by transmission electron microscopy in Rous sarcoma virus transformed chick embryo chondrocytes. Peculiar "glove finger" invaginations were found to take origin from the ventral membrane at sites of close contact; they were directed toward the center of the cell perpendicularly from the substratum. These new structures may be the sites where the local release of proteases takes place at the side of cell-to-substratum adhesion in podosome-bearing cells. The cytoplasmic face of glove finger invaginations and of the plasma membrane at cell-to-cell contact is lined by thick accumulations of microfilamentous material. PMID- 2557980 TI - The formation of multivesicular structures in the adipose cells of chick embryos. AB - Observation of the cytogenesis of adipose tissue of the chick embryo revealed a quantity of multiversicular structures (MVs) which were found in the intercellular space. Some of them were attached to the adipocytes and others were independently located in the intercellular space. The origin of those MVs appeared to be part of the degenerating mitochondria. Centrally located vesicles and vacuoles in degenerating mitochondria formed a group of short tubules and vacuoles which protruded through the cytoplasmic membrane or bulged out at the edge of the cytoplasmic process. The MVs then spread over the cytoplasmic membrane and finally were discharged from the cell surface as in the manner of apocrine secretion. An invisible barrier between the mass of vesicles and the rest of the cytoplasmic structures appeared to segregate the extruding MVs from the intercellular components such as ribosomes, microtubules, and microfilaments. PMID- 2557981 TI - Oxygen radicals in CNS damage. AB - The products of univalent reduction of oxygen, superoxide anion radical, hydrogen peroxide, and the hydroxyl radical, are capable of causing cellular damage and death. They are, therefore, logical candidates as mediators of vascular and parenchymal injury in the central nervous system (CNS). This paper reviews the sources of oxygen radicals in the CNS, their effects on cerebral vessels and on brain and spinal cord parenchyma, and the evidence which implicates oxygen radicals in various pathological conditions of the CNS. PMID- 2557982 TI - On the mechanism of the Mn3(+)-induced neurotoxicity of dopamine:prevention of quinone-derived oxygen toxicity by DT diaphorase and superoxide dismutase. AB - Dopamine (DA) is rapidly oxidized by Mn3(+)-pyrophosphate to its cyclized o quinone (cDAoQ), a reaction which can be prevented by NADH, reduced glutathione (GSH) or ascorbic acid. The oxidation of DA by Mn3+, which appears to be irreversible, results in a decrease in the level of DA, but not in a formation of reactive oxygen species, since oxygen is neither consumed nor required in this reaction. The formation of cDAoQ can initiate the generation of superoxide radicals (O2-.) by reduction-oxidation cycling, i.e. one-electron reduction of the quinone by various NADH- or NADPH-dependent flavoproteins to the semiquinone (QH.), which is readily reoxidized by O2 with the concomitant formation of O2-.. This mechanism is believed to underly the cytotoxicity of many quinones. Two electron reduction of cDAoQ to the hydroquinone can be catalyzed by the flavoprotein DT diaphorase (NAD(P)H:quinone oxidoreductase). This enzyme efficiently maintains DA quinone in its fully reduced state, although some reoxidation of the hydroquinone (QH2) is observed (QH2 + O2----QH. + O2-. + H+; QH. + O2----Q + O2-.). In the presence of Mn3+, generated from Mn2+ by O2-. (Mn2+ + 2H+ + O2-.----Mn3+ + H2O2) formed during the autoxidation of DA hydroquinone, the rate of autoxidation is increased dramatically as is the formation of H2O2. Furthermore, cDAoQ is no longer fully reduced and the steady-state ratio between the hydroquinone and the quinone is dependent on the amount of DT diaphorase present. The generation of Mn3+ is inhibited by superoxide dismutase (SOD), which catalyzes the disproportionation of O2-. to H2O2 and O2. It is noteworthy that addition of SOD does not only result in a decrease in the amount of H2O2 formed during the regeneration of Mn3+, but, in fact, prevents H2O2 formation. Furthermore, in the presence of this enzyme the consumption of O2 is low, as is the oxidation of NADH, due to autoxidation of the hydroquinone, and the cyclized DA o-quinone is found to be fully reduced. These observations can be explained by the newly-discovered role of SOD as a superoxide:semiquinone (QH.) oxidoreductase catalyzing the following reaction: O2-. + QH. + 2H+----QH2 + O2. Thus, the combination of DT diaphorase and SOD is an efficient system for maintaining cDAoQ in its fully reduced state, a prerequisite for detoxication of the quinone by conjugation with sulfate or glucuronic acid. In addition, only minute amounts of reactive oxygen species will be formed, i.e. by the generation of O2-., which through disproportionation to H2O2 and further reduction by ferrous ions can be converted to the hydroxyl radical (OH.). Absence or low levels of these enzymes may create an oxidative stress on the cell and thereby initiate events leading to cell death. PMID- 2557983 TI - Heterocyclic quinones. XVI. Pharmacomodulation in the series of 11H-indolo[3,2 c]quinolinediones: synthesis, cytotoxicity and antitumor activity of 3 substituted 11H-pyrido[3',4':4,5]pyrrolo[3,2-c]quinoline-1,4-diones. AB - With the aim of obtaining new antitumor drugs more active than previously described 11H-indolo[3,2-c]quinoline-1,4-diones and 7,8,9,10-tetrahydro-11H indolo[3,2-c]quinoline-1,4-diones, the synthesis and activities of a series of 3 substituted 11H-pyrido[3',4':4,5]pyrrolo[3,2-c]quinoline-1,4-diones and of 7,8,9,10-tetrahydro-11H-pyrido-[3',4':4,5]pyrrolo[3,2-c] quinoline-1,4-diones were studied. Some quinones were more cytotoxic in vitro towards L1210 leukemia cells but were not active in vivo towards murine P388 leukemia. PMID- 2557984 TI - Studies on 2',3'-dideoxy-2',3'-didehydropyrimidine nucleosides. II. N4-benzoyl 2',3'-dideoxy-2',3'-didehydrocytidine as a prodrug of 2',3'-dideoxy-2',3' didehydrocytidine (DDCN). AB - N4-Benzoyl-2',3'-dideoxy-2',3'-didehydrocytidine (Bz-DDCN) was synthesized as a novel prodrug of 2',3'-dideoxy-2',3'-didehydrocytidine (DDCN), which is a reverse transcriptase inhibitor and is considered to be a potential anti-acquired immunodeficiency syndrome agent. Chemical and enzymatic regeneration of DDCN from the prodrug has been investigated in both in vitro and in vivo experiments. Bz DDCN regenerated DDCN under basic conditions (greater than pH 8), while cleavage of the N-glycosidic linkage and production of N4-benzoylcytosine were observed under acidic conditions (less than pH 6). DDCN was enzymatically regenerated from the prodrug in the presence of several enzyme preparations, including human plasma. DDCN and Bz-DDCN were intravenously administered to mice and the plasma concentrations of DDCN and the prodrug were measured. Though DDCN levels following direct DDCN administration decreased exponentially with a half-life of 14.5 min, the plasma levels of DDCN following the prodrug administration were sustained above 2 microM for over 3 h. PMID- 2557985 TI - Pyrimidine 5'-nucleotidase deficiency: improved detection of carriers. AB - The activities of pyrimidine 5'-nucleotidase (P5N) and the nucleotide pools in the erythrocytes from 19 members of a Dutch family with P5N deficiency were measured. In the erythrocytes of 5 (out of 6) apparent heterozygotes (based on P5N activities), an increased amount of UDP-N-acetylhexosamines was found. This increase was also found in the erythrocytes of 2 (out of 3) questionable heterozygotes (P5N activity below normal range, but not below normal mean--2 X SD) and not in the erythrocytes of family members with a normal P5N activity nor in erythrocytes from healthy donors. We conclude that analysis of the ribonucleotide patterns, in combination with determination of P5N activity, allows a more accurate diagnosis of heterozygosity for P5N. PMID- 2557986 TI - Plasma ACTH responses to cortisol infusion are similar in patients with primary hypoadrenalism and patients studied some years after bilateral adrenalectomy for Cushing's syndrome. AB - It has been suggested that the first-phase feedback of steroid feedback on ACTH secretion is abnormal in Cushing's disease patients studied after adrenalectomies. We have performed hydrocortisone infusions (100 mg over 2 h) in 15 patients who had previously undergone bilateral adrenalectomy for Cushing's disease. None had had any pituitary-directed therapy and none had a pituitary macroadenoma. Their responses were compared with a group of seven hypoadrenal patients. A significant rise in serum cortisol was observed between 5 and 10 min in both groups. The first significant fall in ACTH occurred between 30 and 45 min in both groups. There was no abnormality of first-phase feedback in the Cushing's group and our results do not therefore support previous speculation that such an abnormality provides evidence for a higher central nervous system (CNS) aetiology of Cushing's disease. PMID- 2557987 TI - Elevated serum levels of calcitonin gene-related peptide (CGRP) but no evidence for CGRP gene expression in non-small cell lung carcinomas. AB - Calcitonin gene-related peptide (CGRP) is a hormone formed by alternative post transcriptional processing of the calcitonin gene. It is a neuropeptide localized to discrete regions of the central nervous system (CNS) and in nerve fibres associated with blood vessels. It is also expressed in medullary carcinomas of the thyroid and lung carcinoma cell lines. The latter finding suggests a possible value for CGRP as tumour marker in lung carcinomas. In this investigation of 22 patients undergoing operation for lung tumours, pre and post-operative levels of serum CGRP were measured. Preoperative as well as postoperative serum CGRP levels were significantly elevated when compared to age-matched normals. However, no evidence could be found for CGRP gene expression in tumour tissue from the same patients as judged by immunocytochemistry or in-situ hybridization using CGRP cRNA probes. CGRP has been localized to nerve fibres in relation to pulmonary blood vessels and has been shown to be a potent vasodilator. These findings, and the absence of evidence for synthesis in tumours, as opposed to cell lines derived from lung carcinomas, suggests that the lack of post-operative normalization of serum CGRP concentrations may be related to physiological changes in cardiovascular haemodynamics following surgery. Elevated pre-operative serum CGRP levels may also reflect a consequence of the lung carcinoma leading to increased release of CGRP from sites in the vasculature yet to be determined, but does not indicate synthesis de novo and secretion of CGRP by the tumours. PMID- 2557988 TI - Acute dexamethasone suppression of ACTH secretion stimulated by human corticotrophin releasing hormone, AVP and hypoglycaemia. AB - In order to obtain more insight into the mechanisms regulating endogenous ACTH secretion in humans we studied the inhibitory effect of acute i.v. dexamethasone administration on ACTH release under various conditions. Six male volunteers were subjected to six different protocols. After combined i.v. injection of 100 micrograms corticotrophin releasing hormone (CRH) and 100 micrograms growth hormone releasing hormone (GRH) there was the expected rise in ACTH (area under the curve, 1053 +/- 204 (SE) (pmol/l) min) and cortisol (59788 +/- 10098 (nmol/l) min) rise which was suppressed by prior i.v. injection of 2 mg dexamethasone (ACTH: 444 +/- 63 (pmol/l) min; cortisol: 28528 +/- 2152 (nmol/l) min). Insulin hypoglycaemia (IH) led to a more pronounced ACTH and cortisol rise compared with CRH (6307 +/- 817 (pmol/l) min and 82080 +/- 21934 (nmol/l) min, respectively) which was not completely suppressed by prior pretreatment with dexamethasone (ACTH, 580 +/- 103 (pmol/l) min; cortisol: 55649 +/- 5821 (nmol/l) min). Combined AVP/CRH injection (10 IU/100 micrograms) after pretreatment with dexamethasone (344 +/- 41 (pmol/l) min for ACTH; 32832 +/- 3173 (nmol/l) min for cortisol) could not reproduce the ACTH secretion following IH after pretreatment with dexamethasone (579 +/- 103 (pmol/l) min for ACTH and 55649 +/- 5821 (nmol/l) min for cortisol). In all subjects a saline control with 2 mg dexamethasone was performed. These findings confirm the acute inhibitory effect of glucocorticoids on CRH-stimulated ACTH secretion. Since CRH-induced ACTH secretion is almost completely abolished by administration of dexamethasone the ACTH rise following IH after dexamethasone can not be mediated by endogenous CRH alone. Moreover, since the addition of AVP to CRH (after dexamethasone suppression) could not reproduce the ACTH rise during IH after dexamethasone pretreatment, an additional, yet unknown factor stimulating ACTH secretion may be involved. In the same protocols, no significant difference could be observed comparing IH and GRH induced GH secretion (4948 +/- 1172 (mU/l) min vs 3596 +/- 820 (mU/l) min, NS); furthermore, in contrast to results obtained by chronic steroid administration, acute i.v. dexamethasone pretreatment did not affect IH or GRH-induced GH secretion (4110 +/- 666 (mU/l) min vs 2916 +/- 462 (mU/l) min, NS). The GRH stimulated GH secretion (3596 +/- 820 (mU/l) min) was not suppressed by prior intravenous treatment with dexamethasone (2916 +/- 504 (mU/l) min, NS). PMID- 2557989 TI - Genome juggling by transposons: Tam3-induced rearrangements in Antirrhinum majus. AB - Transposable elements are well known for their ability to generate large- and small-scale rearrangements of the sequences flanking their insertion sites. These include deletions, inversions, and duplications. Tam3, a transposon from the Snapdragon (Antirrhinum majus), is highly active in the generation of such rearrangements. We have analysed a number of Tam3-induced rearrangements at the nivea (niv) locus by Southern blotting, cloning, and sequence determination. The data obtained from these analyses have led to an understanding of the mechanisms by which these complex alleles were formed. We have shown that the primary rearrangements usually occur without excision of the element and therefore result from aberrant transposition attempts. Subsequent rearrangements may occur on excision of the element. Finally, we suggest how the analysis of such rearrangements may not only provide information about Tam3 transposition but also show how transposon-induced rearrangements may influence the structure and function of the genome as a whole. PMID- 2557990 TI - Characterization of bz1 mutants isolated from mutator stocks with high and low numbers of Mu1 elements. AB - The high frequency of mutations in Mutator stocks of maize is the result of transposition of Mu elements. Nine different Mu elements that share the 220 bp Mu terminal inverted repeats have been described. Mu1 elements have been found inserted into most of the molecularly characterized mutant alleles isolated from Mutator stocks, and most Mutator stocks contain a high number of Mu1 elements (10 60). However, it is clear that additional Mu elements, which share the Mu1 termini but have unrelated internal sequences, can also transpose in Mutator stocks. We were interested in comparing the mutation frequency and type of elements that inserted into a particular locus when Mutator stocks with differing numbers of Mu1 elements were utilized. Furthermore, previous studies with Mu induced mutations have demonstrated that the element that inserted most frequently was Mu1. Therefore, to try to obtain Mu elements different from Mu1 we utilized a stock that had a low number (3-6) of Mu1 elements as well as a Mutator stock with a more typical number of Mu1 elements (20-60). Utilizing both stocks, we isolated numerous mutants at one gene, Bronze 1 (Bz1), and compared the type of elements inserted. In this paper we report that both the high and low Mu1 stocks produced bz1 mutants at frequencies characteristic of Mutator stocks, 6.6 and 4.3 x 10(-5), respectively. We describe the isolation of 20 bz1 mutations, and the initial molecular characterization of eight unstable mutations: two from the high Mu1 stock and six from the low Mu1 stock. The six alleles isolated from the low Mu1 stock appear to contain deleted Mu1 elements, and the two alleles isolated from the high Mu1 stock contain elements very similar to Mu1. When the mutants from the low Mu1 stocks were examined, it was found that the Mu1-related elements increased from 3-6 copies to 9-20 copies in one generation. The high number of Mu1-related elements was maintained in subsequent outcrosses. This spontaneous activation and amplification of Mu1-related elements occurred in at least 1% of the low Mu1 plants. PMID- 2557991 TI - Genetic and molecular characterization of a-mrh-Mrh, a new mutable system of Zea mays. AB - A new allele of the maize A1 gene, a gene required for anthocyanin pigment biosynthesis, was identified in a genetic stock exhibiting a high frequency of chromosome breakage at the second microspore mitosis. This allele, a-mrh, is unstable in both somatic and germinal tissue when an independent locus, Mrh, is present in the genome. a-mrh was molecularly cloned, and a 246 bp DNA insertion with characteristics of a transposable element was identified within the fourth exon of the gene. Southern blot analysis of germinal derivatives of a-mrh suggests that the DNA insert rMrh is excised from the locus when a wild-type phenotype is restored. Genetic crosses with components of other two-element mutable systems of maize failed to induce mutability. We therefore conclude that rMrh is a member of a new, two-element transposon system of maize. The genetic and molecular characteristics of the elements involved are discussed with respect to stress-activated transposition, response of an element to developmental signals, and a possible new role of plant transposons in gene evolution. PMID- 2557992 TI - Developmental and genetic aspects of Mutator excision in maize. AB - The regulation of excision of Mu elements of the Mutator transposable element family of maize is not well understood. We have used somatic instability of Mu receptor elements from the Bronze 1 and Bronze 2 loci to monitor the frequency and the timing of excision of Mu elements in several tissues. We show that spot size in the aleurone of a bz2::mu1 stock varies between one to approximately 256 cells. This indicates that excision events begin eight divisions prior to full aleurone differentiation and end after the last division of the aleurone. We show that excision is equally biased for late events in all other tissues studied. A locus on chromosome 5 has been identified that affects spot size, possibly by altering the timing of Mu excision. Using somatic excision as an assay of Mutator activity, we found that activity can change in small sectors of the tassel; however, there are no overall activity changes in the tassel during the period of pollen shedding. We also report the recovery of germinal revertants for the bz1::mu1 and bz2::mu1 alleles. One of these revertant alleles was characterized by Southern blot analysis and found to be similar to the progenitor of the mutable allele. PMID- 2557993 TI - The mechanisms of action of nonsteroidal antiinflammatory drugs. AB - In mammalian species and in the oldest of multicellular animal forms, NSAIDs inhibit cell activation, apparently in the absence of effects on PG biosynthesis. Thus, an alternative hypothesis can be proposed to account for the antiinflammatory effects of these drugs. Clearly, at low doses aspirin and most of the newer NSAIDs inhibit the biosynthesis of PGs from arachidonic acid, and stable PGs have been shown to mediate fever, hyperalgesia, vasodilation (edema), and several interleukin-1-dependent responses. At high doses, however, aspirin, sodium salicylate, and the newer NSAIDs (at antiinflammatory doses) inhibit non PG-dependent processes, such as the activity of a variety of enzymes, proteoglycan synthesis by chondrocytes, transmembrane ion fluxes, and chemoattractant binding. These effects are most likely due to the capacity of aspirin-like drugs to insert into the lipid bilayer of plasma membranes, where they disrupt normal signaling events and protein-protein interactions. The ability of NSAIDs to thereby inhibit the activation of inflammatory cells such as the neutrophil may contribute to the antiinflammatory properties of this class of drugs. PMID- 2557995 TI - [Migrating multiple mononeuritis and nonsystemic angitis]. AB - Multiple mononeuritis of migrating nature at onset occurs in a variety of disease conditions. "Migrant sensory neuritis" without a systemic underlying disorder described by Wartenberg (1958) is the most distinctive form of this type of neuropathy. Its pathomechanism is not uniform, but angiopathy has been suggested by Matthews et al. (1981). In this report, we describe five cases of sensory dominant multiple mononeuritis with migrating nature and without systemic visceral involvement. The patients consisted of four females and one male between 26 and 71 years of age. All showed recurrent episodes of sensory involvement along the distal branches of the cutaneous nerve. The patients presented with sudden onset of numbness or pain radiating along the cutaneous branches of the involved nerves often followed by persistent sensory deficit. In mild episodes, the sensory symptom with numbness almost completely subsided. The frequency of these episodes ranged from 7 times in 6 months to 6 times in 9 years. The clinical manifestations were similar or identical to those of migrant sensory neuritis reported by Watenberg and Matthews et al. Cranial nerve involvement and less frequent episodes in the present series differed from those in the previous reports. Laboratory examinations did not disclose any underlying disorder, suggesting systemic collagen vascular disease. Sural nerve biopsy study in two patients revealed vasculitis in small or medium-sized arteries in the epineurium as well as reduced population of large myelinated fibers. Small myelinated fibers were slightly increased in number, probably to regenerating fibers. Sensory action potentials were low or not evoked in the nerves examined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557996 TI - [Acute autonomic and sensory neuropathy associated with galactorrhea-amenorrhea syndrome and intractable anorexia]. AB - Acute autonomic and sensory neuropathy (AASN), one subtype of acute pandysautonomia, in which dorsal root ganglia and autonomic ganglia are involved is uncommon. Little is so far known on central nervous system involvement in AASN. In the present paper we described a rare case of AASN associated with the central nervous system manifestations such as galactorrhea-amenorrhea syndrome and intractable anorexia. A 30-year-old woman rapidly developed burning pain and numbness in her arms and legs as well as orthostatic syncope. She had severe anorexia and no no menstruation from onset. On physical examination, she was emaciated. There was marked orthostatic hypotension with tachycardia. Skin was dry. Moderate galactorrhea was detected. Neurological examination showed prominent paresthesia and dullness of superficial sensation, predominantly to pinprick and thermal stimuli, segmentally over the neck, occipital scalp, and extremities. Deep sensation was intact. She had no weakness or ataxia. Deep tendon reflexes were almost normal. NCV and SEP were normal, while EEG was abnormal. Sural nerve biopsy demonstrated axonal degeneration with the loss of myelinated, predominantly in small-caliber fibers, and unmyelinated fibers. The levels of HVA and MHPG in CSF were decreased. The autonomic nervous function tests revealed postganglionic dysfunction. alpha-adrenergic system was predominantly impaired, while beta-adrenergic system was relatively preserved. The endocrinological studies demonstrated mild or moderate elevation of PRL basal value and hyper-response of PRL and LH for TRH and LH-RH loading test, which suggested disorder of the hypothalamo-hypophysial system. Cranial MRI showed moderate dilatation of the 3rd ventricle.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557997 TI - [Subacute sensory neuropathy manifestated by tonic pupils associated with small cell carcinoma of the lung]. AB - Only a few cases of carcinomatous neuropathy with tonic pupils have been reported. In the present paper we described a 53-year-old woman with subacute sensory neuropathy who had presented with bilateral tonic pupils. She noticed numbness over the medial aspect of the left thigh in March, 1988, and then developed dysesthesia over the left forearm and hand, mild weakness of left upper and lower extremities and urinary disturbance. Neurological symptoms were subacutely progressive and she was bed-ridden in May. She was admitted to our hospital in June, 1988. On examination, she had body-weight loss of 6 kg during the last six months and general status was otherwise unremarkable. She had anisocoria; the left pupil was larger in daylight than the right, while smaller in dim light. The left pupil scarcely reacted to light, but promptly constricted to near vision. The right pupil constricted normally to light and near vision. An instillation of 0.0625% pilocarpine solution showed supersensitive response of both pupils. An instillation of 1.25% epinephrine solution demonstrated mild dilation of both pupils. Thus, it was conceivable that she had postganglionic ciliary nerve damage characteristic of tonic pupil as well as the lesion of sympathetic nerve innervating pupillary dilator. She had severe sensory ataxia and pseudoathetosis of the hands. Weakness was mild to moderate in extremities. Almost all deep tendon reflexes were absent. All modalities of sensation, particularly on deep sense, were severely involved with sea-level-type distribution below Th7, and over C2 to C3 regions on the left side.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2557998 TI - New perspectives for diagnosis of nodular goiter by technetium-thallium subtraction scanning. AB - To improve the scintigraphic differential diagnosis of thyroid nodules, Tc-99m pertechnetate-Tl-201 Cl subtraction scintigraphy (Tc-Tl subtraction scanning) was performed in 106 patients with various histologically proven thyroid nodules. Results were evaluated by comparison with surgical findings and preoperative ultrasonographic results. Thyroid nodules were more successfully detected by Tc Tl subtraction scanning than by either Tc-99m pertechnetate or Tl-201 Cl scintigraphy alone. Detection of nodules was further improved when images recorded by the three methods were integrally observed (sensitivity 83%, accuracy 89%), with detectability approaching that of ultrasonography. False-positive or negative Tc-Tl subtraction scans were obtained principally when multiple nodules were present (7 cases) or when no discrepancy existed between the accumulations of Tc-99m pertechnetate and Tl-201 Cl (18 cases). Most colloid nodules exhibited irregular margins, heterogeneous internal accumulations of Tl-201 Cl, and distorted shapes on Tc-Tl subtraction scans, while the majority of adenoma were oval-shaped with smooth margins. Carcinoma were characterized by homogeneous internal accumulation of Tl-201 Cl and distorted shapes. PMID- 2557994 TI - Possible mechanisms of action of the antiphospholipid binding antibodies. AB - If antibodies directed against the phospholipid itself play any part in the pathogenesis of the thrombotic events, the very ubiquitous nature of the antigen dictates that no single pathogenetic mechanism can be implicated. The majority of in vivo studies recently performed have failed to show that antiphospholipid (aPL) antibodies bind cultured endothelial cells, or that they affect prostacyclin production by these cells. Previously postulated actions showing defective fibrinolysis and impairment of coagulation inhibitors (protein C, thrombomodulin and antithrombin III) have yet to be substantiated. Studies have thus far failed to demonstrate any significant binding of aPL antibodies to intact platelets, although binding to disrupted platelets has been documented. PMID- 2557999 TI - Scintigraphic depiction of an insulinoma by I-131 metaiodobenzylguanidine. AB - Scintigraphy with I-131 metaiodobenzylguanidine (MIBG) was effective in depicting a pancreatic insulinoma in a patient suffering from intermittent hypoglycemia. This observation widens the range of neuroendocrine tumors that take up to I-131 MIBG and supports the concept that many tumors of the amine precursor uptake and decarboxylation system may be imaged in this way. PMID- 2558001 TI - [Calcium channel as molecular target of drug action]. AB - The authors discuss the recent acquisitions concerning the function and the molecular structure of Ca++ channels in excitable cells. Furthermore, they focus on the clinical pharmacology of Ca++ entry blockers in order to give more insight into the pharmacological basis for their therapeutic use. PMID- 2558000 TI - Stimulation of erythrocyte and renal Na+,K+-adenosine triphosphatase activity by antidigoxin antibody in normal rats. AB - 1. A circulating ouabain-like factor which inhibits the Na+,K(+)-pump has been implicated in volume-expanded states. To assess the role of this putative factor in normovolaemic rats, we measured erythrocyte and renal Na+,K(+)-adenosine triphosphatase activity after the infusion of a mixture of high-affinity digoxin binding Fab fragments (Digibind) capable of removing digoxin from pump sites. 2. Compared with either saline (vehicle) or sheep immunoglobin G, infusion of the antidigoxin antibody caused a moderate increase of Na+,K(+)-adenosine triphosphatase activity in the erythrocyte (saline 348 +/- 12; immunoglobulin G 339 +/- 16; antidigoxin antibody 432 +/- 22 nmol h-1 mg-1; P less than 0.005 by analysis of variance) and a larger increase in the renal cortex (saline 9.7 +/- 0.9; immunoglobulin G 9 +/- 1.4; antidigoxin antibody 24.3 +/- 1.8 mumol h-1 mg 1; P less than 0.0005 by analysis of variance) without a change in blood pressure. 3. These results are consistent with the presence of a digoxin-like inhibitor of the Na+,K+-pump in normal rats. PMID- 2558002 TI - Outpatient diagnosis and treatment of AIDS-related pulmonary infections. AB - A large number and variety of organisms can infect the lungs of persons with AIDS. An abnormal chest X-ray associated with pulmonary symptoms should trigger a vigorous search for an infectious agent. In many cases, if the patient is ambulatory, the workup can proceed on an outpatient basis. Pneumocystis carinii pneumonia is the most common infection and also one of the most readily treatable illnesses that these patients encounter. A careful and considered diagnostic approach is necessary to conserve the time and resources of both patients and healthcare providers. PMID- 2558003 TI - Long-term starvation in Xenopus laevis Daudin--III. Effects on enzymes in several tissues. AB - 1. Adult, female Xenopus laevis were subjected to 12 months of starvation. 2. Starvation resulted in a continuous reduction in the activity of both hepatic and renal glucose-6-phosphate dehydrogenase. 3. Fructose-1,6-diphosphatase was significantly reduced at months 10 and 12 in the liver, and at months 4, 10, and 12 in the kidney. 4. Pyruvate kinase activity of muscle and liver decreased during the experimental period whereas the renal enzyme remained essentially unchanged. 5. Both hepatic and renal glutamate-pyruvate transaminase (GPT) and hepatic glutamate-oxaloacetate transaminase (GOT) showed a reduction of activity after 2 and 4 months of starvation followed by an increase in GPT but not in GOT. PMID- 2558004 TI - RECPAM: a computer program for recursive partition amalgamation for censored survival data and other situations frequently occurring in biostatistics. II. Applications to data on small cell carcinoma of the lung (SCCL). AB - The RECPAM methodology previously presented in part I (A. Ciampi et al., Comput. Methods Programs Biomed. 26 (1988) 239-256) is applied to the analysis of survival data on small cell carcinoma of the lung (SCCL). It is shown how RECPAM can help answer the following questions which occur frequently in the analysis of clinical data: Is it possible to find a classification of patients with a certain disease into distinct prognostic groups? Given a covariate of special interest, does it have an independent prognostic significance even after confounding is taken into account? Does the prognostic significance of a covariate of special interest vary across patient subgroups? For the SCCL data, a prognostic classification is obtained and the tumor marker LDH is treated as a variable of special interest. Many features of RECPAM are illustrated, including, among others, Forward and Backward (Pruning) Stopping Rules, treatment of missing data, and use of several dissimilarity measures. PMID- 2558005 TI - Dietary fiber and health. AMA council on Scientific Affairs. AB - During the last 18 years, considerable research has been conducted on the role of dietary fiber in health and disease. Interest was stimulated by epidemiologic studies that associated a low intake of dietary fiber with the incidence of colon cancer, heart disease, diabetes, and other diseases and disorders. Dietary fiber is not a single substance. There are significant differences in the physiological effects of the various components of dietary fiber. A Recommended Dietary Allowance for dietary fiber has not been established. However, an adequate amount of dietary fiber can be obtained by choosing several servings daily from a variety of fiber-rich foods such as whole-grain breads and cereals, fruits, vegetables, legumes, and nuts. PMID- 2558006 TI - Evidence of the non-infectivity of herpes simplex viral particles in Trichomonas vaginalis. AB - Fan et al., employing an indirect immunofluorescent antibody (IFA) technique, reported the presence of herpes simplex virus type 2 (HSV-2) in Trichomonas vaginalis. However, using the same method, we found that the protozoa showed autofluorescence immediately after acetone fixation. In order to demonstrate the non-infectivity of HSV-2 in T. vaginalis, several other methods were performed in this study. Trichomonads were experimentally incubated together with HSV-2 and examined after immunofluorescent staining procedures. Organisms without any contact with HSV-2 were used as control. Results obtained from both vital stained and methanol-fixed organisms failed to show any fluorescence. Data from DNA fluorochrome staining, immunoblot, electron microscopy and viral titer assay were in accordance with the results of the IFA method. No obvious difference between the freshly isolated and the long term-cultured organisms could be detected by these methods. All strains of T. vaginalis investigated by this study failed to show the presence of intracellular HSV-2. PMID- 2558007 TI - Detection of lactobacilli and their interaction with clostridia in human gastrointestinal tracts and in vitro. AB - Culture counts of aerobic lactobacilli in the feces of vegetarians ([8.1 +/- 0.7] of log10 bacteria per gram dry weight) were higher than those of meat consumers ([5.2 +/- 0.1] of log10 bacteria per gram dry weight). Co-culture of Lactobacillus acidophilus and Clostridium perfringens in various media revealed that an interaction between these two species. The pH was the most important factor controlling their relative growth, which might explain the relative abundance of lactobacilli in feces of men and animals on vegetarian diets. Lactobacilli but not clostridia were detected in feces of newborn infants. These findings are discussed in relation to health. PMID- 2558009 TI - Peripheral neuropathy in spinal cord injured patients. AB - We performed electrophysiological studies on the lower limbs of a group of clinically complete spinal cord injured patients. Our findings indicated a frequent asymmetric sensory and motor nerve involvement at proximal and distal sites. We suggest that mechanical compression and traction is one of etiological factors for such findings. PMID- 2558008 TI - Isolated symptomatic peripheral neuropathy in type 1 insulin-dependent diabetes mellitus. AB - Diabetic neuropathy is probably the most frequent of the chronic complications of diabetes, and is usually found in association with diabetic retinopathy and/or nephropathy. We report seven patients with long-standing insulin-dependent diabetes mellitus in whom symptomatic peripheral neuropathy was the first and only documented complication. The diagnosis of peripheral symmetrical neuropathy was based on the presence of symptoms and abnormal physical findings, confirmed with abnormal electrophysiological and/or vibratory and thermal threshold measurements. Diabetic retinopathy and nephropathy were absent. We conclude that in some type 1 insulin-dependent diabetic patients, similar to what has been reported in type 2 non-insulin-dependent diabetes, peripheral neuropathy may be the first chronic complication to become manifest. This observation provides additional evidence to suggest that each of the diabetic complications may have a different pathogenic mechanism. PMID- 2558010 TI - Crosslinking of cytochrome c and cytochrome b5 with a water-soluble carbodiimide. Reaction conditions, product analysis and critique of the technique. AB - A water soluble carbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), has been used to crosslink horse heart cytochrome c and trypsin solubilized bovine liver microsomal cytochrome b5. The reaction was conducted under a variety of solution conditions, and the products were purified by a combination of gel filtration and ion-exchange chromatography. Under all conditions of pH, ionic strength, EDC/protein ratio and reaction time that were studied, multiple 1:1 crosslinked complexes were observed with no evidence of a single, dominant species. Acetate, which is often used as a quencher of such reactions, was found to increase the complexity of the reaction products, presumably through EDC-promoted coupling to cytochrome c. Hydroxylamine treatment of the crosslinked complexes, a procedure frequently used to reverse EDC modification of tyrosyl residues, did not reduce the number of crosslinked components observed. The cytochrome b5 heme group was readily extracted from each of the 1:1 crosslinked complexes by standard techniques, so the crosslinking of heme propionate 7 with Lys79 of cytochrome c that might have been anticipated on the basis of molecular graphics modeling [Salemme, F.R. (1976) J. Mol. Biol. 102, 563-568] was not evident from this analysis. Analysis of HPLC tryptic peptide maps produced from crosslinked complexes revealed reduced specificity of trypsin in hydrolysis of EDC-crosslinked protein-protein complexes and unsatisfactory resolution of crosslinked or branched peptides. Nevertheless, it was possible to demonstrate that residues 52-72 of cytochrome b5, a region predicted to be critical to interaction with cytochrome b5 [Salemme, F.R. (1976) J. Mol. Biol. 102, 563-568] was absent from all peptide maps of 1:1 cytochrome c.cytochrome b5 complexes. Based on these results and a review of the literature involving EDC crosslinking of electron transfer proteins, we conclude that the techniques available for specific protein hydrolysis and separation of crosslinked peptides are not adequate to permit routine unambiguous identification of crosslinking sites in carbodiimide-crosslinked complexes. PMID- 2558011 TI - Restriction of poliovirus RNA translation in a human monocytic cell line. AB - The infection of the human monocytic cell line U-937 by poliovirus was characterized by a low level of virus production and a slow progression of the cytopathic effect. Infection took place in greater than 99% of the cells as revealed by a limiting dilution assay. No viral protein synthesis was evident in the infected U-937 cells when analyzed by polyacrylamide gel electrophoresis. However, a low level of poliovirus RNA translation was detected by immunofluorescence analysis using a mixture of polyclonal antisera against non structural proteins. Although there was only a low level of viral protein synthesis, a gradual accumulation of viral mRNA took place in U-937 cells as revealed by RNA blot analysis. Similar results were obtained when the erythroleukemic cell line K-562 was used as a host cell for poliovirus. RNA extracted from infected U-937 cells was efficiently translated in rabbit reticulocyte extracts giving rise to a pattern of viral polypeptides similar to that detected when virion-purified RNA was the template used for the in vitro translation assay, suggesting that the poliovirus RNA present in infected U-937 cells is functional. The existence in U-937 cells of a discriminatory mechanism which differentially interferes with poliovirus RNA translation is discussed. PMID- 2558012 TI - Poly(A) polymerase from Vigna unguiculata seedlings. A bifunctional enzyme responsible for both poly(A)-polymerizing and poly(A)-hydrolyzing activities. AB - Poly(A)-specific ribonuclease was co-purified with poly(A) polymerase from Vigna unguiculata seedlings. Both activities were separated into two forms (enzymes I and II) by a final hydrophobic column chromatography. The enzyme I preparation, which was homogeneous as examined by SDS/PAGE, had both poly(A) polymerase and poly(A)-specific ribonuclease activities. The antibody raised to the enzyme I preparation precipitated both enzyme activities. These indicate that a single polypeptide (Mr 63,000) is responsible for both poly(A)-polymerizing and poly(A) hydrolyzing activities. The poly(A)-specific ribonuclease was a 3'-exonuclease specific to single-stranded poly(A), forming 5'AMP as the sole reaction product. The hydrolytic activity required either Mn2+ or Mg2+ with different optimum concentrations, whereas the polymerizing activity required Mn2+ but not Mg2+. ATP and PPi had little or no effect on the poly(A)-specific ribonuclease activity. PMID- 2558013 TI - Regulation of protein phosphatase-1G from rabbit skeletal muscle. 1. Phosphorylation by cAMP-dependent protein kinase at site 2 releases catalytic subunit from the glycogen-bound holoenzyme. AB - The glycogen-associated form of protein phosphatase-1 (PP-1G) is a heterodimer comprising a 37-kDa catalytic (C) subunit and a 161-kDa glycogen-binding (G) subunit, the latter being phosphorylated by cAMP-dependent protein kinase at two serine residues (site 1 and site 2). Here the amino acid sequence surrounding site 2 has been determined and this phosphoserine shown to lie 19 residues C terminal to site 1 in the primary structure. The sequence in this region is: (sequence; see text) At physiological ionic strength, phosphorylation of glycogen bound PP-1G was found to release all the phosphatase activity from glycogen. The released activity was free C subunit, and not PP-1G, while the phospho-G subunit remained bound to glycogen. Dissociation reflected a greater than or equal to 4000-fold decrease in affinity of C subunit for G subunit and was readily reversed by dephosphorylation. Phosphorylation and dephosphorylation of site 2 was rate-limiting for dissociation and reassociation of C subunit. Release of C subunit was also induced by the binding of anti-site-1 Fab fragments to glycogen bound PP-1G. At near physiological ionic strength, PP-1G and glycogen concentration, site 2 was autodephosphorylated by PP-1G with a t0.5 of 2.6 min at 30 degrees C, approximately 100-fold slower than the t0.5 for dephosphorylation of glycogen phosphorylase under the same conditions. Site 2 was a good substrate for all three type-2 phosphatases (2A, 2B and 2C) with t0.5 values less than those toward the alpha subunit of phosphorylase kinase. At the levels present in skeletal muscle, the type-2A and type-2B phosphatases are potentially capable of dephosphorylating site 2 in vivo within seconds. Site 1 was at least 10-fold less effective than site 2 as a substrate for all four phosphatases. In conjunction with information presented in the following paper in this issue of this journal, the results substantiate the hypothesis that PP-1 activity towards the glycogen metabolising enzymes is regulated in vivo by reversible phosphorylation of a targetting subunit (G) that directs the C subunit to glycogen--protein particles. The efficient dephosphorylation of site 2 by the Ca2+/calmodulin-stimulated protein phosphatase (2B) provides a potential mechanism for regulating PP-1 activity in response to Ca2+, and represents an example of a protein phosphatase cascade. PMID- 2558014 TI - Regulation of protein phosphatase-1G from rabbit skeletal muscle. 2. Catalytic subunit translocation is a mechanism for reversible inhibition of activity toward glycogen-bound substrates. AB - The glycogen-associated form of protein phosphatase-1 (PP-1G) comprises a 37-kDa catalytic (C) subunit and a 161-kDa glycogen-binding (G) subunit. In the preceding paper in this issue of the journal we showed that the C subunit is released from PP-1G in response to phosphorylation of the G subunit by cAMP dependent protein kinase. We now show that at 0.15-0.2 M KCl the phosphorylase phosphatase activity of glycogen-bound PP-1G is 5-8 times higher than that of released C subunit or unbound PP-1G, which are strongly inhibited at these ionic strengths. The activity of glycogen-bound PP-1G towards glycogen synthase was about 5-fold higher than that of released C subunit at 0.15M KCl. Studies with glycogen-bound substrates and myosin P-light chain (which does not interact with glycogen) indicated that PP-1G activity is only enhanced compared to free C subunit at near physiological ionic strength and when both PP-1G and substrate are glycogen-associated. The inhibition by increasing ionic strength and enhanced activity upon binding to glycogen reflected changes in K'm, but not Vmax. From the determined specificity constant, k'cat/K'm approximately 4 x 10(6) s-1 M-1, it was calculated that at physiological levels of glycogen-bound PP-1G (200 nM) and phosphorylase (70 microM), dephosphorylation of the latter could occur with a half time of 15 s, sufficient to account for inactivation rates in vivo. The much higher catalytic efficiency of glycogen-bound PP-1G toward the glycogen metabolising enzymes at physiological ionic strength compared to free C subunit substantiates the role of PP-1G in the regulation of these substrates, and establishes a novel mechanism for selectively regulating their phosphorylation states in response to adrenalin and other factors affecting phosphorylation of the G subunit. PMID- 2558015 TI - Biochemical characterization of phospholipase D activity from human neutrophils. AB - We have found a phospholipase D activity in the postnuclear fraction of human neutrophils, employing phosphatidylinositol as exogenous substrate. This phospholipase D activity was assessed by both phosphatidate formation and by free inositol release in the presence of 15 mM LiCl in the reaction mixture and in the absence of Mg2+ ions to prevent inositol-1-phosphate phosphatase activity. To assess further the phospholipase D activity, we studied its capacity to catalyze a transphosphatidylation reaction, as a unique feature of the enzyme. It was detected as [14C]phosphatidylethanol formation when the postnuclear fraction was incubated with [14C]phosphatidylinositol in the presence of ethanol. The phospholipase D showed a major optimum pH at 7.5 and a minor one at pH 5.0. Neutral and acid phospholipase D activities were differentially located in subcellular fractionation studies of resting neutrophils, namely in the cytosol and in the azurophilic granules, respectively. Neutral phospholipase D required Ca2+ ions to the active, whereas the acid enzyme activity was Ca2(+)-independent. The neutral phospholipase D activity showed a certain specificity for phosphatidylinositol, as it was able to hydrolyze phosphatidylinositol at a much higher rate than phosphatidylcholine, in the absence and in the presence of different detergents. This neutral phospholipase D activity behaved as a protein of high molecular mass (350-400 kDa) by gel filtration chromatography. Moreover, neutral phospholipase D activity was detected in the postnuclear fraction of human monocytes, by measuring free inositol release from phosphatidylinositol as exogenous substrate, under the same experimental conditions as those used with neutrophils. The enzyme displayed similar specific activities in both cell types as well as the same degree of activation after cell stimulation with the calcium ionophore A23187. These results demonstrate the existence of two phospholipase D activities with different pH optima and intracellular location in human neutrophils. Furthermore, these results suggest that this phospholipase D can play a role in signal-transducing processes during cell stimulation in human phagocytes. PMID- 2558016 TI - Identification of a new heparin-binding protein localized within chick basement membranes. AB - A protein with a molecular mass of 19 kDa has been purified to homogeneity from 11-day-old chick embryos using a procedure involving chromatography on heparin - Sepharose, immunoaffinity resin and C4 reversed-phase. Indirect immunofluorescence studies, using polyclonal and monoclonal antibodies raised against this protein, indicate that it is essentially localized within the basement membranes in early embryonic tissues. After the 18th day of embryonic life and in post-hatched chicken, this protein could only be detected in some eye basement membranes. It appears to be bound to heparan sulfate chains of the proteoglycan present in these structures. Thus, the protein exhibits similar properties to those previously described for fibroblast growth factors (FGF), such as heparin affinity, molecular mass and localization in the basement membranes. In contrast, this protein is present in much larger amounts than FGFs, at least in 11-day-old embryos. Furthermore, the first 17 amino acid residues of the N-terminal sequence show that it does not strictly correspond to any previously described protein. PMID- 2558017 TI - Direct photoaffinity labeling of leukotriene binding sites. AB - Due to their conjugated double bonds the leukotrienes themselves are photolabile compounds and may therefore be used directly for photoaffinity labeling of leukotriene binding sites. Cryofixation eliminates unspecific labeling taking place in solution by photoisomers and photodegradation products of leukotrienes. After fixation of receptor ligand interactions by shock-freezing of the samples, irradiation-induced highly reactive excited states and/or intermediates can form covalent bonds with the respective binding site in the frozen state. After cryofixation of a solution of albumin incubated with [3H8]leukotriene E4, irradiation at 300 nm resulted in time-dependent incorporation of radioactivity into the protein. Photoaffinity labeling of rat as well as of human blood serum with [3H8]leukotriene E4 after cryofixation revealed that only one polypeptide with an Mr of 67,000 was labeled. This polypeptide was identified as albumin. Photoaffinity labeling of rat liver membrane subfractions enriched with sinusoidal membranes resulted in the labeling of a polypeptide with an apparent Mr of 48,000, whereas no polypeptide was predominantly labeled in the subfraction enriched with canalicular membranes. Photoaffinity labeling of isolated hepatocytes disclosed different leukotriene E4 binding polypeptides. In the particulate fraction of hepatocytes a polypeptide with an apparent Mr of 48,000 was labeled predominantly, whereas in the soluble fraction several polypeptides were labeled to a similar extent. One of these, with an apparent Mr of 25,000, was identified as subunit 1 of glutathione transferases by immunoprecipitation. The method of direct photoaffinity labeling in the frozen state after cryofixation using leukotrienes as photoactivatable compounds, as exemplified by leukotriene E4, may be most useful for the identification and characterization of various leukotriene binding sites, including receptors, leukotriene-metabolizing enzymes, and transport systems. PMID- 2558018 TI - Two siblings with vitamin-D-dependent rickets type II: no recurrence of rickets for 14 years after cessation of therapy. AB - Rickets in a 3-year-old boy and his 1-year-old sister, both with alopecia, was cured by treatment with 50,000 IU of vitamin D2 daily for 2 years and did not recur within 14 years after cessation of therapy. A diagnosis of vitamin-D dependent rickets type II was made in these patients at the ages of 20 and 18 years based on the findings that 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] did not inhibit DNA biosynthesis in phytohaemagglutinin-stimulated lymphocytes and that cultured skin fibroblasts showed impaired nuclear uptake and normal cytosol binding of [3H]1,25(OH)2D3. Surprisingly, the serum 1,25(OH)2D levels of these patients were high and their serum 24,25-dihydroxyvitamin D levels were low, although neither patient showed any symptoms except alopecia. The presence of vitamin D metabolite imbalances in the absence of rickets in these patients might be explained by differences in sensitivity to 1,25(OH)2D3 of bone formation and vitamin D metabolism. In addition, changes of sensitivity to treatment with vitamin D derivatives might be a consequence of differentiation of target cells. From the present findings, it is suggested that in this disease treatment with a sufficient dose of vitamin D derivatives should be initiated in the active phase of rickets. PMID- 2558019 TI - Nedocromil sodium cream in the treatment of atopic dermatitis. PMID- 2558020 TI - Disparate functions of I-A and I-E molecules on B cells as evidenced by the inhibition with anti-I-A and anti-I-E antibodies of polyclonal B cell activation. AB - Polyclonal differentiation of unprimed B cells into IgM-producing cells induced by lipopolysaccharide (LPS) or T cell-derived lymphokine B151-TRF2 has been shown to contain a process of I-A-restricted B-B cell interaction, so that the B cell responses are inhibited by monoclonal antibodies (mAb) specific for I-A molecules. On the other hand, the B cell responses are also inhibited by anti-I-E mAb, although I-E molecules are not involved in such B-B cell interaction. In this study, we examined the mechanism underlying the anti-I-E-mediated inhibition of the B cell responses. The B cell responses induced by LPS or B151-TRF2 were inhibited by either anti-I-A or anti-I-E mAb added on day 0 over a 5-day culture period, whereas when added on day 3 the responses were inhibited only by anti-I-E mAb and not by anti-I-A mAb. To gain insight into the mechanism underlying the anti-I-E-mediated inhibition, we prepared monovalent Fab and divalent F(ab')2 fragments of anti-I-A and anti-I-E mAb and examined their effects on the B cell responses. We found that the B cell responses were inhibited by the F(ab')2 but not Fab fragment of anti-I-E mAb, whereas the Fab fragment of anti-I-A mAb still gave effective inhibition. The F(ab')2 but not Fab fragment of anti-I-E mAb induced increases in cyclic AMP (cAMP) levels in B cells, whereas the undigested anti-I-A mAb did not induce such increases. Furthermore, adenylate cyclase inhibitors, which inhibit cellular cAMP accumulation, circumvented the B cell responses inhibited by anti-I-E but not anti-I-A mAb. Thus, these results indicate that the anti-I-E-mediated inhibition of the B cell responses requires increases in intracellular cAMP levels induced by cross-linking of I-E molecules. In contrast, anti-I-A mAb inhibits the B cell responses without cross-linking of I-A molecules and cAMP accumulation. These results reinforce a unique function of I-A molecules as restriction elements in the Ia-restricted B-B cell interaction. PMID- 2558021 TI - Immunological alterations inducible by mercury compounds. III. H-2A acts as an immune response and H-2E as an immune "suppression" locus for HgCl2-induced antinucleolar autoantibodies. AB - In responder mouse strains repeated injections of subtoxic doses of HgCl2 induce formation of antinuclear autoantibodies (ANA) and antinucleolar autoantibodies (ANolA). Others have shown that responsiveness to HgCl2-induced formation of ANA and ANolA is linked to H-2. Here, we extend these studies to a variety of mouse strains not tested previously. After confirming that strain B10.S (H-2s) is a high responder we have shown that strains B10.D2 (H-2d) and B10.BR (H-2k) are nonresponders. By comparing a panel of strains carrying appropriate intra-H-2 recombinant haplotypes derived from d, k and s, we were able to map responsiveness to As. Interestingly, among four strains all of which were As, and thus responsive, only the two H-2E- ones, B10.S and B10.RSD2, were high responders whereas the two H-2E+ ones, B10.HTT and B10.S(9R), were significantly less responsive. Thus, the genetics of HgCl2-induced autoantibody formation follow the rules established for immune responses to a variety of different antigens in that expression of H-2E "suppressed" the response. PMID- 2558022 TI - Derivation of a T cell hybridoma variant deprived of functional T cell receptor alpha and beta chain transcripts reveals a nonfunctional alpha-mRNA of BW5147 origin. AB - We have isolated a variant of the DO-11.10.7 mouse T cell hybridoma which does not express functional T cell receptor alpha/beta chains. This variant, denoted 58 alpha-beta-, can be used as a recipient for T cell receptor alpha/beta gene transfer experiments to obtain cell lines which express only the products of the transfected alpha/beta genes at their surfaces. In the process of characterizing the defects affecting the 58 alpha-beta-T cell receptor genes, we have found that the parental BW5147 thymoma has undergone a previously unnoticed V alpha-J alpha rearrangement. This alpha rearrangement involves a V alpha pseudogene segment and accounts for the high level of alpha-mRNA transcripts present in the BW5147 alpha beta- variant. Knowledge of the existence of this second, albeit nonfunctional, alpha-mRNA in BW5147 is of importance, since it could be, and actually already has been, mistakenly identified (due to partial nucleotide sequencing) in T hybrids as a functionally significant message donated by the normal T cell parent. PMID- 2558023 TI - Structure and expression of the T cell receptor gamma locus in pre-B and early hemopoietic cells. AB - The genetic structure and expression of the T cell receptor (TcR) loci were examined in pre-B and early hemopoietic cells. Thirty-eight percent of Abelson murine leukemia virus-transformed pre-B cell lines were rearranged at TcR gamma. Moreover, many pre-B cell lines were rearranged at two distinct gamma loci, V gamma 1.2 and V gamma 2. The gamma rearrangements in the pre-B cell lines were similar to those observed previously in T cell lines. V + C-containing gamma mRNA was detected in two pre-B cell lines. In all other pre-B and interleukin (IL) 3 dependent lymphoid and myeloid lines examined, smaller C gamma-containing mRNA were detected. These C gamma transcripts were independent of the genetic configuration of the gamma locus. In contrast, the TcR alpha and beta loci were in the germ-line configuration in all non-T cell lines examined and mRNA encoding these loci were not detected. When IL3-dependent lymphoid and myeloid cell lines were transformed to growth factor independence by a non-autocrine mechanism, no mRNA transcripts encoding TcR C gamma were detected. However, TcR C gamma mRNA transcripts were detected in factor-independent cell lines that arose by an autocrine mechanism. The cell cycle expression of C gamma was compared with protooncogenes and other marker genes previously shown to be cell cycle specific. mRNA transcripts encoding C gamma were detected in the highest amounts 4-8 h after IL 3, but not phorbol myristate acetate, addition. A similar time period of expression was observed with ornithine decarboxylase which has been shown to be expressed in G1 phase. These observations indicate that TcR gamma is often rearranged in pre-B cell lines and may be directly regulated by IL3 in IL3 dependent cells. PMID- 2558024 TI - Effects of muscarinic receptor agonists and antagonists on alpha 2-adrenoceptors in rat brain. AB - The specific binding of [3H]clonidine to alpha 2-adrenoceptors on neural membranes isolated from six brain areas was determined with rats treated for various periods of time with the muscarinic agonists, oxotremorine or pilocarpine, or with the muscarinic antagonists atropine, atropine methyl nitrate, scopolamine and scopolamine methyl bromide. Administration of pilocarpine, 10 mg/kg, twice daily i.p. for 1 and 14 days increased markedly the number of alpha 2-adrenoceptors on neural membranes from all six brain areas. In contrast, oxotremorine, 0.3 mg/kg, twice daily i.p., for 7 days decreased the number of alpha 2-adrenoceptors on membranes from all brain areas except the brainstem and caudate nucleus. Both atropine and scopolamine increased the density of alpha 2-adrenoceptors in specific brain areas. Neither atropine methyl nitrate nor scopolamine methyl bromide had an appreciable effect upon the specific binding of [3H]clonidine to neural membranes from most brain areas. PMID- 2558025 TI - NMDA receptor blockade in the periaqueductal grey prevents stress-induced analgesia in attacked mice. AB - Microinjections of N-methyl-D-aspartate (NMDA, 0.1 and 1.0 nmol) into the periaqueductal grey (PAG) of the mouse resulted in potential antinociception. In a social conflict situation, attacked mice exhibited a marked analgesia that was prevented by prior injection of the competitive NMDA antagonist, AP-7 (2.0 nmol) or naloxone (6.0 nmol) into the PAG and also by i.p. injection of the non competitive NMDA antagonist, MK-801 (33 nmol). These results demonstrate that NMDA receptors are involved in endogenous analgesic mechanisms activated by stress. PMID- 2558026 TI - Direct autoradiographic localization of adenosine A2 receptors in the rat brain using the A2-selective agonist, [3H]CGS 21680. AB - The regional distribution of adenosine A2 receptors in the rat brain was determined using the A2-selective agonist ligand [3H](2-p carboxyethyl)phenylamino)-5'-N-carboxamidoadenosine (CGS 21680) by quantitative receptor autoradiography. [3H]CGS 21680 binding was highly localized in the striatal region of the rat brain with the greatest density of binding found in the caudate-putamen, nucleus accumbens and olfactory tubercle. Additionally, lower levels of binding were also found in the globus pallidus. No significant amounts of [3H]CGS 21,680 binding were detected in other brain regions. This localization of brain adenosine A2 receptors was markedly different from the known regional distribution of A1 receptors which are highly concentrated in cerebellum, hippocampus, thalamus and cortex. The present results provide further evidence for a specific contribution of adenosine in the modulation of central neurotransmission. PMID- 2558027 TI - Antagonism of methoxyflurane-induced anesthesia in rats by benzodiazepine inverse agonists. AB - Injection of the partial benzodiazepine inverse agonist Ro15-4513 (1-32 mg/kg i.p.) or nonconvulsant i.v. doses of the full benzodiazepine inverse agonist beta CCE immediately following cessation of exposure of rats to an anesthetic concentration of methoxyflurane significantly antagonized the duration of methoxyflurane anesthesia as measured by recovery of the righting reflex and/or pain sensitivity. This antagonism was inhibited by the benzodiazepine antagonist Ro15-1788 at doses which alone did not alter the duration of methoxyflurane anesthesia. In addition, high-dose Ro15-4513 pretreatment (32 mg/kg) antagonized the induction and duration of methoxyflurane anesthesia but was unable to prevent methoxyflurane anesthesia or affect the induction or duration of anesthesia induced by the dissociative anesthetic ketamine (100 mg/kg). These findings indicate that methoxyflurane anesthesia can be selectively antagonized by the inverse agonistic action of Ro15-4513 and beta-CCE. PMID- 2558028 TI - Parasagittal zonation of GABA-B receptors in molecular layer of rat cerebellum. PMID- 2558029 TI - GABA preincubation of rat brain sections increases [3H]GABA binding to the GABAA receptor and compromises the modulatory interactions. AB - Receptor autoradiography has been employed to investigate the effect of gamma aminobutyric acid (GABA) preincubation on the interaction of the GABAA receptor with its ligands. [3H]GABA (50 nM) binding to the GABAA receptors is increased by 60% compared to control sections after GABA (100 microM) preincubation. Receptor autoradiography shows that the increase is more pronounced in certain brain areas. The allosteric interactions between the GABA and benzodiazepine recognition sites were also examined. An increase in [3H]GABA (50 nM) binding to rat brain sections by co-incubation with the benzodiazepine, flunitrazepam (FNZ) has been observed autoradiographically. This effect has been quantitated in several brain regions; the overall brain increase in [3H]GABA binding induced by 1 microM FNZ was 20%. The increase in [3H]FNZ (1 nM) binding by co-incubation with GABA has also been observed autoradiographically, and the effect quantitated in four brain regions. The overall brain increase in [3H]FNZ binding induced by 100 microM GABA was 34%. After GABA preincubation these allosteric responses are significantly reduced in size. The increase in the [3H]GABAA binding as a consequence of GABA preincubation appears to reflect an increase in receptor affinity for [3H]GABA with no significant change in the maximum number of binding sites. We suggest that GABA preincubation converts the GABAA receptor to a higher affinity desensitised receptor conformation. PMID- 2558030 TI - Comparative study of visual inspections and microbiological sampling in premises manufacturing and selling high-risk foods. AB - The possible relationship between the results of a microbiological sampling programme and visual inspections carried out in local food-manufacturing premises was examined. Using five main parameters - overall appearance, personal hygiene, risk of contamination, temperature control, and training and education - a visual inspection rating score was established for each of the premises. A variety of high-risk processed foods, and specimens from hands, wiping cloths and environmental swabs were examined. The results from two study periods indicated that there was an overall poor agreement between microbiological results and inspection ratings. On its own, neither sampling nor visual assessment reliably monitored the performance of the premises. A combined approach, using selective microbiological examination to support a system of standardized inspections, is suggested for monitoring food hygiene standards in premises selling high-risk foods. PMID- 2558031 TI - The survival patterns of selected faecal bacteria in tropical fresh waters. AB - The survival of various faecal bacteria used as indicators of the faecal contamination of water supplies has been investigated in a tropical environment (Sierra Leone). Isolates representing the thermotolerant coliform (TtC) and faecal streptococcus (FS) groups, Clostridium perfringens and Salmonella spp. were studied over a 48 h period of immersion in water from three different sources. Survival patterns varied according to source type, but some general observations were made: a portion of the TtC group was apparently capable of substantial regrowth; FS organisms died off at a faster rate than TtC organisms initially, but survived longer; vegetative cells of C. perfringens died off rapidly; and Salmonella spp. could survive for as long as the other faecal organisms tested. The implications of results for the analysis of tropical waters for faecal contamination are discussed. PMID- 2558033 TI - The time course of the humoral immune response to rhinovirus infection. AB - The specific humoral immune response of 17 volunteers to infection with human rhinovirus type 2 (HRV-2) has been measured both by neutralization and by ELISA. Six volunteers who had HRV-2-specific antibodies in either serum or nasal secretions before HRV-2 inoculation were resistant to infection and illness. Of the remaining 11 volunteers who had little pre-existing HRV-2-specific antibody, one was immune but 10 became infected and displayed increases in HRV-2-specific antibodies. These antibodies first increased 1-2 weeks after infection and reached a maximum at 5 weeks. All six resistant volunteers who had high pre existing antibody and eight of the volunteers who became infected maintained their HRV-2-specific antibody for at least 1 year. At this time they were protected against reinfection. Two volunteers showed decreases in HRV-2-specific antibodies from either serum or nasal secretions. They became infected but not ill after HRV-2 inoculation 1 year later. PMID- 2558032 TI - Lymphoproliferative response to fusion proteins of human papillomaviruses in patients with cervical intraepithelial neoplasia. AB - The cell-mediated immune response (CMI) to E6 and E4 fusion proteins of human papillomavirus type 16 (HPV-16), E6 fusion protein of HPV-18, and to control proteins similarly produced, was analysed in 29 patients with cervical intraepithelial neoplasia (CIN) and in 15 age-matched laboratory personnel using a lymphocyte proliferation assay (LPA). Compared to controls without any added proteins, a positive response (stimulation index greater than 2.0) to the highly purified E6 control protein was found in only one patient. Positive responses to the E4 control protein which contained beta-galactosidase were noted in three patients and two controls. With control proteins as baseline, the lymphocytes from nine patients (28%) and three laboratory personnel (20%) responded to at least on HPV fusion protein after 7 days in culture. Stimulation indices were low in both groups with a range of 2.06-4.69 and the difference in incidence of positive responses between the groups is not significant. Proliferative responses to HPV-1 and HPV-2 virion antigens were noted in 6/23 (26%) of the patients and 2/15 (18%) of the other group. No correlation between responsiveness and degree of dysplasia or presence of koilocytes was found in the patient group. The relevance of the low proliferative responses is discussed. PMID- 2558034 TI - Restricted variability of a 17 nucleotide stretch within the 5'-noncoding region of poliovirus genome. AB - The outbreak of poliomyelitis in Finland in 1984 was caused by a wild strain of poliovirus 3 with uncommon molecular and antigenic properties. We prepared a synthetic oligonucleotide probe complementary to nucleotides 494-510 in the 5' noncoding part of the genome of a representative strain of the outbreak. This short nucleotide stretch was found to be relatively well conserved within the outbreak and uncommon among 82 independent poliovirus isolates. It may thus be a useful marker for screening isolates to identify those requiring more detailed genetic comparison. The sequences of the corresponding region of the genome are known for 32 separate poliovirus strains and 3 coxsackie B virus strains and show 6 fully conserved nucleotides that could assume a constant hairpin-loop position in a hypothetical secondary structure of the RNA. This could explain the persistence of a particular 17 nucleotide sequence for 40 years in nature in this highly variable region of the poliovirus genome. PMID- 2558035 TI - Approaches to the molecular epidemiology of rickettsioses. AB - This review deals with the developments of molecular approaches to the investigation of rickettsial disease epidemiology. The data presented include changes in the incidence and geographic distribution of endemic rickettsioses. Use of the DNA restriction enzyme technique, in combination with DNA probe analysis, for the molecular genetic differentiation of tick spotted fever--and typhus fever--group rickettsiae and correlation between with the analysis of polypeptide composition of the above group of rickettsiae are discussed. The data are presented on progress in the identification of various Coxiella burnetii strains as a result of restriction analysis of plasmid DNA as well as chromosomal DNA in combination with DNA probe. New and detailed characteristics of classified and newly isolated strains of rickettsiae and Coxiella burnetii revealed by molecular genetic differentiation techniques are discussed. New identification techniques using DNA probes in combination with restriction analysis of chromosomal from rickettsiae and both plasmid and chromosomal DNA from Coxiella burnetii are considered to have good prospects for future use in epidemiological assessment. The establishment of reference file banks containing restriction endonuclease data on the available typical and atypical strains of rickettsiae and Coxiella burnetii is suggested. PMID- 2558036 TI - Structural analysis of proviruses in additional hamster tumour cell lines transformed by provirus II rescued from XC cells and definition of a new cell line harbouring amplified proviruses. AB - Additional 18 Syrian hamster tumour cell lines induced independently with the virus rescued from XC cells (provirus type II) have been analysed by restriction mapping. Taking into account the already published results of an analysis of other six tumour cell lines, altogether 24 tumour cell lines were characterized. In 62.5% of these lines a complete proviral unit has been integrated; 20.8% of the lines harbour altered proviral structures where the 3' end of the gag gene and the entire pol gene had been deleted; 8.3% of the lines carry both such a deleted provirus and a complete provirus. Apart from the already published cryptic provirus found in H-19 cells, no such provirus has been found in another cell line. Therefore the appearance of the cryptic provirus represents an infrequent event (4.16%). Similarly, the significant amplification of proviruses accompanied with a rearrangement in the env gene was found only in one tumour cell line. H-42, in the genome of which five proviral units are accommodated. The probable arrangement of amplified proviruses and the mechanism of their genesis are discussed. PMID- 2558037 TI - Colonic lymphoid cell subsets and epithelial HLA-DR antigens in familial polyposis coli. AB - Although there has been some evidence suggesting that immunological mechanisms are involved in the pathophysiology of familial polyposis coli (FPC), there has not been any report as to whether there is any abnormality of lymphoid cell subsets as background, whether polyps (adenomas) show different lymphoid cell subsets from the normal mucosa, or whether HLA-DR antigens are expressed on the epithelia of adenomas. Lymphoid cell subsets (CD5, CD4, CD8, IgA1, IgA2, IgM, IgD, IgG, and IgE positive cells) in the lamina propria, and HLA-DR antigens on the epithelia were studied in 7 patients with FPC. From each patient, 2 specimens were obtained from both the normal (non-polypoid) area and the polyp. Normal colonic mucosa, taken from 15 patients with conventional polyps or colorectal cancer, served as the normal control mucosa. Lymphoid cell subsets and HLA-DR antigens were identified by indirect immunoperoxidase staining using mouse anti human monoclonal antibodies. In the normal area of FPC, lymphoid cell subsets were similar to those of normal control mucosa except for an increase in IgD positive (IgD+) cells. However, definite alterations were observed in the polyp. There were significant increases in the number of CD4+ and IgG+ cells, and in the sum of five classes of Ig+ cells compared to the normal area or normal control mucosa. HLA-DR antigens were not expressed in the normal control mucosa or in the normal areas, and only on the epithelia of the polyp in 5 out of 7 specimens (71%). These results clearly demonstrate that immunological reactions are involved in FPC polyps. PMID- 2558038 TI - Production of leukotriene B4 in parenchymal and sinusoidal cells of the liver in rats treated simultaneously with D-galactosamine and endotoxin. AB - A study was conducted to investigate production rate of leukotriene B4 (LTB4) in parenchymal and sinusoidal liver cells of rats with acute hepatic failure (AHF). AHF was induced by simultaneous administration of D-galactosamine (GalN) and endotoxin (LPS), and parenchymal as well as sinusoidal liver cells were isolated by collagenase perfusion method. Following preincubation for 15 min, isolated cellular fractions were incubated with Ca-ionophore (2 microM) for 5 min, and levels of LTB4 in culture media before and 5 min after addition of Ca-ionophore were analyzed by HPLC. Following results were obtained: The production rate of LTB4 was found to be the highest in Kupffer cells (7.2ng/10(6) cells/5 min), followed by endothelial cells (1.1), stellate cells (0.2) and parenchymal cells (not detectable). The production rate of LTB4 in both Kupffer cells and endothelial cells was found to reach a maximum in the fraction isolated 60 min after administration of GalN and LPS. Treatment with AA861, one of the selective inhibitors of 5-lipoxygenase, was shown to reduce the production of LTB4 in Kupffer cells to 53% at 10(-7)M and above 99% at higher than 10(-5)M. In conclusion, the majority of LTB4 generated in the liver of rats with AHF was found to be synthesized in Kupffer cells and, to a lesser extent, in endothelial cells, and the enhanced production of LTB4 was found to be greatly inhibited by treatment with AA861. PMID- 2558039 TI - A female case of type VIII glycogenosis who developed cirrhosis of the liver and hepatocellular tumor. AB - The case of a 17-year-old female with a rare form of type VIII glycogenosis who developed cirrhosis of the liver and hepatocellular tumor is reported. Laparoscopy showed a tumor 50 mm in diameter in the lower portion of the right lobe of the liver. The tumor was biopsied under ultrasonic guidance, and tentatively diagnosed as adenomatous hyperplasia. The patient was also diagnosed as having type VIII glycogenosis (phosphorylase kinase deficiency). PMID- 2558040 TI - Age-related changes in rat brain muscarinic receptors and beta-adrenoreceptors. AB - 1. In experiments on 2-, 10- and 22-month old rats, it was found that the Bmax values for muscarinic receptors and beta-adrenoreceptors increased in the cerebral cortex, striatum and hippocampus of 10-month old rats as compared to those in 2-month old rats. 2. The Bmax values for both receptor types significantly decreased in the same brain structures of 22-month old rats as compared to those in 10-month old rats. In the striatum and hippocampus of 22 month old rats the binding capacity decreased as compared also to those in 2 month old rats. 3. In the hypothalamus there was also a tendency towards increasing the binding capacity of 10-month old rats and towards decreasing the binding capacity of 22-month old animals only for muscarinic receptors. The beta max of beta-adrenoreceptors remained unchanged in all age groups studied. 4. The receptor affinity of both receptor types was in most cases unaltered with advancing age. The Kd values were slightly increased only in the striatum and hippocampus of 22-month old rats as compared to 10-month old rats. 5. The role of age for the changes in the activity of brain muscarinic and beta-adrenoreceptor systems is discussed. PMID- 2558041 TI - Pentylenetetrazole-induced seizure activity produces an increased release of calcium from endoplasmic reticulum by mediating cyclic AMP-dependent protein phosphorylation in rat cerebral cortex. AB - 1. To determine the involvement of the convulsant agent pentylenetetrazole (PTZ) in intracellular calcium release in neurons, its effect on stored calcium in the endoplasmic reticulum of rat cortical neurons was tested. 2. Intraperitoneal injection of PTZ caused marked release of calcium from the endoplasmic reticulum which was similar to that observed when cortical slices were incubated with this convulsant. 3. Superfusion of dibutyryl cAMP and isobutylmethylxanthine to the cortical slices mimicked PTZ-induced calcium release from this reservoir. A similar effect was observed under depolarizing conditions brought about by either an elevation of extracellular K+ concentration or addition of veratridine. 4. Isoquinolinesulfonamide, a protein kinase inhibitor, reduced PTZ-stimulatory effect of calcium release and blocked the cAMP-induced calcium release. 5. Intracellular cAMP level was enhanced at about 3-fold by both intraperitoneal injection of PTZ and its superfusion. 6. These findings are taken to suggest that PTZ may release stored calcium in the endoplasmic reticulum by mediating a cAMP dependent protein phosphorylation in cortical neurons. PMID- 2558042 TI - Age-related changes in [3H]prazosin binding and phosphoinositide hydrolysis in rat ventricular myocardium. AB - 1. We investigated, using rats at 2, 12 and 24 months of age, the effect of aging on ventricular alpha 1-adrenoceptor responsiveness by measuring the [3H]prazosin binding, [3H]inositol phosphate formation, the cholesterol/phospholipid ratio and membrane viscosity. 2. The density of specific [3H]prazosin binding sites on membranes was significantly reduced with advanced age, and the binding affinity for [3H]prazosin was significantly increased. 3. Norepinephrine (NE)-induced formation of [3H]inositol phosphates in slices was also reduced with advanced age. However, the EC50 for NE stimulation of [3H]inositol phosphates was significantly higher when compared to those at adult ages. 4. The cholesterol content in membranes was significantly decreased with advanced age; concomitantly, a decrease in the cholesterol/phospholipid ratio was observed. The membrane viscosity as measured by using 1,6-diphenyl-1,3,5-hexatriene significantly decreased with advanced age. 5. These results suggest that the altered cholesterol/phospholipid ratio and/or the membrane viscosity during aging may account for the increase in the binding affinity for [3H]prazosin and/or the reduced responsiveness of aged ventricular myocardium to NE. PMID- 2558043 TI - Inhibition of tetrodotoxin-sensitive plateau sodium current by amiodarone in guinea pig cardiac muscles. AB - 1. The acute effects of amiodarone (AM), a potent antiarrhythmic drug, on tetrodotoxin (TTX)-sensitive component of action potentials of Purkinje fibers from guinea pig were studied by use of conventional microelectrode techniques, and compared the findings with the results obtained in the papillary muscle. 2. The present study showed that the action potentials of Purkinje fibers (PF) were more sensitive to AM, compared to those of papillary muscle. 3. Acute exposure (30 min) to 4.4 x 10(-5) M AM led to a depression of plateau potential of PF action potential, but the drug did not affect the total action potential duration. 4. TTX (2-4 x 10(-6) M) shortened the PF action potential duration at all levels of repolarization. 5. The depression of PF plateau potential in the presence of AM was, at least in part, involved in a decrease in TTX-sensitive plateau sodium current, because no further depression of the plateau potential was observed by addition of TTX in the presence of AM. PMID- 2558044 TI - Retinoic acid receptor expression vector inhibits differentiation of F9 embryonal carcinoma cells. AB - Expression vectors have been constructed for a region of the human retinoic acid receptor-alpha (hRAR-alpha) and transferred into F9 embryonal carcinoma (EC) cells. When the vectors are overexpressed in F9 cells, clones can be selected for resistance to retinoic acid-induced differentiation. This effect is obtained even when the hRAR-alpha region is expressed as a beta-galactosidase fusion protein. Using the beta-galactosidase component of the fusion protein as a marker, overexpression of the fusion protein has been correlated with the retinoic acid resistance effect. The clones resistant to retinoic acid no longer exhibit the normal retinoic acid induction of endo B cytokeratin, laminin B-1, and tissue plasminogen activator mRNAs observed with normal F9 cells. Retinoic acid induction of type IV alpha-1 collagen and Hox-1.3 RNAs is observed with these clones. When transfected with a thyroid receptor DNA-binding sequence (TRE)/thymidine kinase promoter/luciferase construct, the retinoic acid-resistant clones do not yield the same retinoic acid-induced level of luciferase obtained with F9 cells. It is hypothesized that the RAR vectors are interfering with endogenous RAR(s) in a dominant-negative manner to inhibit retinoic acid-induced differentiation of F9 EC cells. PMID- 2558045 TI - Four factors are required for 3'-end cleavage of pre-mRNAs. AB - We reported previously that authentic polyadenylation of pre-mRNAs in vitro requires at least two factors: a cleavage/specificity factor (CSF) and a fraction containing nonspecific poly(A) polymerase activity. To study the molecular mechanisms underlying 3' cleavage of pre-mRNAs, we fractionated CSF further and show that it consists of four separable subunits. One of these, called specificity factor (SF; Mr, approximately 290,000), is required for both specific cleavage and for specific polyadenylation and thus appears responsible for the specificity of the reaction. Although SF has not been purified to homogeneity, several lines of evidence suggest that it may not contain an essential RNA component. Two other factors, designated cleavage factors I (CFI; Mr, approximately 130,000) and II (CFII; Mr, approximately 110,000), are sufficient to reconstitute accurate cleavage when mixed with SF. A fourth factor, termed cleavage stimulation factor (CstF; Mr, approximately 200,000), enhances cleavage efficiency significantly when added to a mixture of the three other factors. CFI, CFII, and CstF do not contain RNA components, nor do they affect specific polyadenylation in the absence of cleavage. Although these four factors are necessary and sufficient to reconstitute efficient cleavage of one pre-RNA tested, poly(A) polymerase is also required to cleave several others. A model suggesting how these factors interact with the pre-mRNA and with each other is discussed. PMID- 2558047 TI - Genetic interactions underlying flower color patterns in Antirrhinum majus. AB - Diverse spatial patterns of flower color in Antirrhinum can be produced by a series of alleles of pallida, a gene encoding an enzyme required for pigment biosynthesis. The alleles arose by imprecise excision of a transposable element, Tam3, and we show that they carry a series of deletions involving progressive removal of sequences adjacent to the excision site. This has enabled us to define three cis-acting upstream regions, A, B, and C, which differentially affect the level of pallida expression in distinct areas of the flower. We show further that an unlinked locus, delila, regulates the spatial distribution of pallida transcript. Deletion of regions ABC at the pallida locus uncouples pallida from regulation by delila, whereas deletion of A or AB brings pallida under regulation by delila in a new area of the flower. These results suggest that diverse patterns of pallida expression reflect the different ways in which alleles interact with a prepattern of both common and spatially specific genetic signals in the flower. PMID- 2558046 TI - Phosphorylation and dephosphorylation of a bacterial transcriptional activator by a transmembrane receptor. AB - Signal transduction in the bacterial Omp, Che, and Ntr systems involves the phosphorylation and dephosphorylation of response regulators (OmpR, CheY and CheB, NRI) that share a homologous domain. We show that in the Omp system, the transmembrane sensor EnvZ, catalyzes both the phosphorylation of OmpR and the dephosphorylation of OmpR-P. The phosphorylation reaction proceeds by a mechanism shared with the Ntr and Che kinases, NRII, and CheA. EnvZ can phosphorylate NRI and can stimulate transcription from the glnAp2 promoter, and similarly, CheA can phosphorylate OmpR and can stimulate transcription from the ompF promoter. OmpR-P formed by either CheA or EnvZ is much more stable than CheY-P and NRI-P, but is rapidly hydrolyzed to OmpR and Pi by EnvZ in the presence of ATP, ADP, or nonhydrolyzable analogs of ATP. Because EnvZ is normally a transmembrane receptor with a periplasmic sensory domain, our results suggest that the role of EnvZ may be to control the intracellular concentration of OmpR-P in response to environmental signals. PMID- 2558048 TI - In situ nucleoprotein structure at the SV40 major late promoter: melted and wrapped DNA flank the start site. AB - New in situ probing methods have been developed and used to probe the nucleoprotein structures at the SV40 major late promoter in infected monkey cells. The region that contains the three proximal transcription elements was probed with DNase I and micrococcal nuclease in transcriptionally active, permeabilized cells, and with the single-strand selective reagent KMnO4 in intact cells. The downstream element is included in a region of enhanced DNase I reactivity at 10- to 11-bp intervals for approximately 140 bp, presumably because of DNA wrapping around a specifically positioned nucleosome particle. The two other proximal DNA elements appear to be mostly melted, with a protecting factor bound primarily to the template DNA strand. The protecting factor directly borders the wrapped particle. These observations provide an initial description of parts of the biological transcription machinery and suggest that the SV40 major late promoter elements are part of a higher order nucleoprotein complex that involves wrapped and melted DNA. PMID- 2558049 TI - Searching for pattern and mutation in the Drosophila genome with a P-lacZ vector. AB - A P-element vector has been constructed and used to generate lines of flies with single autosomal P-element insertions. The lines were analyzed in two ways: (1) the identification of cis-acting patterning information within the Drosophila genome, as revealed by a lacZ reporter gene within the P element, and (2) the isolation of lethal mutations. We examined 3768 independent lines for the expression of lacZ in embryos and looked among these lines for lethal mutations affecting embryonic neurogenesis. This type of screen appears to be an effective way to find new loci that may play a role in the development of the Drosophila nervous system. PMID- 2558050 TI - P-element-mediated enhancer detection: a versatile method to study development in Drosophila. AB - We generated and characterized greater than 500 Drosophila strains that carry single copies of a novel P-element enhancer detector. In the majority of the strains, the beta-galactosidase reporter gene in the P-transposon responds to nearby transcriptional regulatory sequences in the genome. A remarkable diversity of spatially and temporally regulated staining patterns is observed in embryos carrying different insertions. We selected numerous strains as markers for different embryonic organs, tissues, and cells. Many of these strains should allow the study of complex developmental processes, such as nervous system development, which have not been convenient to analyze previously. Also, we present genetic evidence that some of the detected regulatory elements control nearby Drosophila genes. In light of our results, we discuss the diversity and complexity of cis-acting regulatory elements in the genome and the general applications of the enhancer detector method for the study of Drosophila development. PMID- 2558051 TI - P-element-mediated enhancer detection: an efficient method for isolating and characterizing developmentally regulated genes in Drosophila. AB - We describe a new approach for identifying and studying genes involved in Drosophila development. Single copies of an enhancer detector transposon, P[1ArB], have been introduced into flies at many different genomic locations. The beta-galactosidase reporter gene in this construct is influenced by a wide range of genomic transcriptional regulatory elements in its vicinity. Our results suggest that a significant proportion of these regulatory sequences are control elements of nearby Drosophila genes. These genes need not be disrupted for their regulatory elements to be identified by P[1ArB]. The P[1ArB] transposon has been designed to facilitate both rapid cloning and deletion analysis of genomic sequences into which it inserts. Therefore, the enhancer detection system is an efficient method of screening for genes primarily on the basis of their expression pattern and then rapidly analyzing those of particular interest at the molecular and genetic levels. PMID- 2558052 TI - CCAAT/enhancer binding protein activates the promoter of the serum albumin gene in cultured hepatoma cells. AB - An expression vector capable of encoding full-length CCAAT/enhancer-binding protein (C/EBP) has been constructed and tested in transient transfection assays for its capacity to activate transcription from the promoter of the serum albumin gene. When tested in cultured hepatoma cells, the C/EBP expression vector achieved potent trans-activation of the albumin promoter. Less substantial activation was observed when the same experiment was conducted using cultured mouse fibroblasts. Expression vectors that encoded defective forms of C/EBP failed to activate the albumin promoter. Moreover, mutated variants of the albumin promoter that lack the C/EBP-binding site failed to be trans-activated. The data are consistent with the interpretation that C/EBP is a bona fide transcription factor. During the course of these experiments it was noted also that C/EBP is more than an order of magnitude less concentrated in cultured hepatoma cells than it is in adult liver cells. Given these findings, we speculate that C/EBP may play a general role in establishing and maintaining the differentiated, nonproliferative state. PMID- 2558053 TI - Loss of Ras activity in Saccharomyces cerevisiae is suppressed by disruptions of a new kinase gene, YAKI, whose product may act downstream of the cAMP-dependent protein kinase. AB - The yeast Saccharomyces cerevisiae contains two functionally redundant genes RAS1 and RAS2, which are homologous to the mammalian ras gene family and are required for vegetative growth. We isolated and characterized five temperature-sensitive alleles of RAS2. In a ras1 strain, these alleles cause growth arrest at the G1 stage of the cell cycle. Revertants capable of growth at the nonpermissive temperature define four recessive, extragenic complementation groups. Suppressors in one complementation group (designated yak1) are particularly intriguing because they appear to alleviate only the growth defect of the temperature sensitive ras mutants and do not show any of the phenotypes, such as heat shock sensitivity or starvation sensitivity, associated with increased production of cAMP. The YAK1 gene has been cloned, and disruptions generated in vitro reveal that it is not essential for growth and that its loss confers growth to a strain deleted for tpk1, tpk2, and tpk3, the structural genes for the catalytic subunit of the cAMP-dependent protein kinase. These results place Yak1 downstream from, or on a parallel pathway to, the kinase step in the Ras/cAMP pathway. Finally, the coding region predicts a protein with significant homology to the family of protein kinases, suggesting that loss of cAMP-dependent protein kinase function can be suppressed by the loss of a second protein kinase. PMID- 2558054 TI - STE12, a protein involved in cell-type-specific transcription and signal transduction in yeast, is part of protein-DNA complexes. AB - The STE12 gene of Saccharomyces cerevisiae is essential for the expression of genes required for mating, such as those involved in pheromone response, and for genes unrelated to mating but regulated by the presence of an adjacent copy of the transposable element Ty1. We show that the STE12 protein is a component of specific DNA-protein complexes that form with transcriptional control elements from Ty1 and the alpha-pheromone receptor gene STE2. Although a sequence involved in pheromone-dependent transcriptional activation is protected in both complexes, competition experiments indicate that the complexes are intrinsically different from each other. We show that another factor involved in cell-type-specific transcription, PRTF/GRM, is a component of the complex with the STE2 fragment but not the Ty1 fragment. We propose that the STE12 product interacts with different transcription factors in different sequence contexts and that PRTF/GRM is one of these factors. PMID- 2558055 TI - Purification of a set of cellular polypeptides that bind to the purine-rich cis regulatory element of herpes simplex virus immediate early genes. AB - Expression of herpes simplex virus type 1 (HSV1) immediate early (IE) genes is activated by a polypeptide component of the mature virion termed viral protein 16 (VP16). Stimulation of IE expression by VP16 operates via two cis-regulatory sequences: TAATGARAT, and the purine-rich hexanucleotide sequence GCGGAA. VP16 does not bind directly to either of the IE cis-regulatory sequences. Rather, these elements appear to represent binding sites for host cell proteins. Herein, we report the purification of a host cell factor that binds to the GCGGAA motif. We show further that this factor is capable of binding in vitro to an oligomerized form of the hexanucleotide sequence GAAACG, which is common to a variety of virus- and interferon-inducible genes. The GAAACG repeats of interferon- and virus-inducible genes, and the GA-rich repeats of HSV1 IE genes confer similar functional properties when appended to the promoter of a heterologous gene. These observations raise the possibility that HSV1 may activate its IE genes in a manner that exploits one of the components used by mammalian cells to combat virus infection. PMID- 2558056 TI - Structure of the human lck gene: differences in genomic organisation within src related genes affect only N-terminal exons. AB - Although cDNA sequences coding for several Rous sarcoma virus Src-related protein tyrosine kinases (PTKs) have been reported for several years, knowledge of the structure and organisation of genes of the src family is still limited. In this work, a detailed structure and organisation of the human lck gene is reported. A 17-kb genomic clone encoding human p56 Lck, a lymphocyte-specific PTK of the Src related subfamily, has been isolated. The human lck gene is organized in 13 exons, one more than in the human cellular (c)-src gene. The twelve coding exons are located in this clone, whereas the putative 5'-noncoding exon is probably located very far upstream from the second exon. Splicing sites for exons 4 to 12, which encode both conserved phospholipase-C-like and catalytic domains of the Src like PTKs, arise exactly at the same position for the human lck, human c-src and c-fgr genes. The only differences concern the splice sites of exons 1' and 2, which encode the unique N-terminal domain of human Lck. These results give further evidence that the different PTKs of the Src-like family have probably evolved through the mechanism of exon shuffling. PMID- 2558057 TI - Initiation of pro-opiomelanocortin mRNA from a normally quiescent promoter in a human small cell lung cancer cell line. AB - We describe the characteristics of pro-opiomelanocortin (POMC) mRNA synthesized by a human small cell lung cancer (SCLC) cell line that secretes a peptide immunoreactive with antibodies to the POMC-derived component, adrenocorticotropin. While no alteration in restriction endonuclease pattern or structure was found for the SCLC-derived pomc gene vs. the previously described human pomc gene cloned from a fetal liver library, Northern-blot analysis of SCLC RNA using pomc-derived probes showed a hybridizing transcript more than 300 nucleotides longer than POMC mRNA isolated from human pituitaries, as well as a pomc-gene-hybridizing mRNA the same length as pituitary-derived transcripts. 5' end mapping and primer extension analyses showed that the novel mRNA species is initiated at a site 371 bp upstream from the 5' end identified for pituitary derived POMC mRNA. We conclude that synthesis of POMC transcripts occurs from an ordinarily quiescent promoter in the SCLC cell line we have studied, as well as from the pomc promoter normally used in pituitary cells. PMID- 2558058 TI - Sequence analysis of the Clostridium cellulolyticum endoglucanase-A-encoding gene, celCCA. AB - The nucleotide sequence of a Clostridium cellulolyticum endo-beta-1,4- glucanase (EGCCA)-encoding gene (celCCA) and its flanking regions, was determined. An open reading frame (ORF) of 1425 bp was found, encoding a protein of 475 amino acids (aa). This ORF began with an ATG start codon and ended with a TAA ochre stop codon. The N-terminal region of the EGCCA protein resembled a typical signal sequence of a Gram-positive bacterial extracellular protein. A putative signal peptidase cleavage site was determined. EGCCA, without a signal peptide, was found to be composed of more than 35% hydrophobic aa and to have an Mr of 50715. Comparison of the encoded sequence with other known cellulase sequences showed the existence of various kinds of aa sequence homologies. First, a strong homology was found between the C-terminal region of EGCCA, containing a reiterated stretch of 24 aa, and the conserved reiterated region previously found to exist in four Clostridium thermocellum endoglucanases and one xylanase from the same organism. This region was suspected of playing a role in organizing the cellulosome complex. Second, an extensive homology was found between EGCCA and the N-terminal region of the large endoglucanase, EGE, from C. thermocellum, which suggests that they may have a common ancestral gene. Third, a region, which extended for 21 aa residues beginning at aa + 127, was found to be homologous with regions of cellulases belonging to Bacilli, Clostridia and Erwinia chrysanthemi. PMID- 2558059 TI - The block to transcription elongation at the SV40 attenuation site is decreased in vitro by oligomers complementary to segments of the attenuator RNA. AB - We have previously reported that a mechanism resembling attenuation in prokaryotes regulates simian virus 40 (SV40) late gene expression. We have suggested that modulation of the attenuator RNA secondary structure is an integral element regulating the elongation block at the attenuation site [Hay et al., Cell 29 (1982) 183-193]. In the present study, oligodeoxyribonucleotides (oligos), 13-19 nucleotides long, were used to probe the involvement of the attenuator RNA secondary structure in the control of elongation block at the SV40 attenuation site. These oligos are complementary to segments of the attenuator RNA suggested to play a role in the regulation of attenuation. The oligos were added to an in vitro transcription reaction containing SV40 transcription complexes, and their effect on transcription through the attenuation site was measured. As predicted, the three oligos caused specific decreases in the elongation block at the SV40 attenuation site. These results provide direct evidence for the involvement of RNA secondary structure in the attenuation mechanism in SV40. PMID- 2558062 TI - [Molecular pharmacology of opioid receptor mechanisms]. AB - The molecular basis of opioid receptor mechanisms was studied in reconstitution experiments using purified or membrane-bound opioid receptors and purified GTP binding proteins (G-proteins). mu-Opioid receptor exclusively purified from rat brains was reconstituted with G-proteins in lipid vesicles. The mu-agonist stimulated the G-protein activity in both G1 or Go-reconstituted vesicles. The stoichiometry revealed that one molecule of mu-receptor is functionally coupled to plural numbers of Gi or Go molecules and that mu-receptor exists in at least two different subtypes, mu i and mu o, separately coupled to Gi and Go, respectively. In addition, when the mu-receptor was phosphorylated by cAMP dependent protein kinase, the mu-agonist-stimulation of G-protein activity disappeared, while the guanine nucleotide-sensitivity of agonist binding was unchanged. These findings suggest that there are independent domains in the receptor which are related to functional coupling to G-protein and to the agonist binding modulation by G-protein. kappa-Opioid receptor agonist inhibited the G protein activity in guinea pig cerebellar membranes. Further experiments revealed that the kappa-opioid receptor is functionally coupled to an inhibition of phospholipase C activity via an inhibition of Gi-activity. Such a receptor mediated inhibition of G-protein activity may be the first demonstration of a signal transduction mechanism. The delta-opioid receptor agonist showed no effect on G-protein activity in guinea pig striatal and rat cortical membranes, while it stimulated it in NG108-15 cells. In all these membranes, the delta-agonist binding was markedly reduced by GTP gamma S in the presence of MgCl2. These findings suggest that delta-receptors in the brain might be coupled to G-protein without signal transduction. PMID- 2558061 TI - Vascular endothelial cell killing by combinations of membrane-active agents and hydrogen peroxide. AB - Previous studies have demonstrated that a number of membrane-active agents are capable of binding to the surface of polymorphonuclear leukocytes (PMN) resulting in an augmentation of superoxide anion and hydrogen peroxide (H2O2) production in response to soluble stimuli. It is now demonstrated that these same membrane active agents can bind to the surface of endothelial cells and enhance their susceptibility to killing by H2O2. Membrane-active agents which are capable of synergizing with H2O2 include cationic proteins, cationic poly-amino acids, lysophosphatides and enzymes which are capable of degrading membrane phospholipids (e.g., phospholipase C, phospholipase A2 and streptolysin S). In each case, treatment of the target cells with the membrane-active agent and H2O2 produces greater damage than the sum of the damage produced by either agent separately. Since inflammatory lesions, particularly sites of bacterial infection, may contain a rich mixture of cationic substances, phospholipases and phospholipid breakdown products, these substances may contribute to the tissue damage observed at sites of inflammation by enhancing endothelial cell sensitivity to PMN-generated H2O2 as well as by augmenting the generation of H2O2 by PMNs. PMID- 2558060 TI - Mutational analysis of the primer RNA template region in the replication origin (oric) of bacteriophage G4: priming signal recognition by Escherichia coli primase. AB - The primase-dependent phage G4 origin of complementary DNA strand synthesis (G4oric) contains three stable stem-loops (I, II, and III) upstream from the initiation point of primer RNA (pRNA). Site-directed mutagenesis was used to introduce alterations into the nucleotide (nt) sequence of the G4oric pRNA template region. Mutations in stem-loop I, that changed the length of the stem and the sequence of the loop, slightly depressed, but did not abolish, G4oric activity. However, functional G4oric activity was destroyed when the sequence containing the starting position of pRNA synthesis was deleted, or when insertions were introduced between the pRNA starting position (5'-CTG-3') and stem-loop I. Reintroducing a CTG as part of a PstI linker close to stem-loop I, however, resulted in recovery of G4oric functional activity. These results suggest that the specific nt sequence, containing 5'-CTG-3', between nt 3994 and 4007, and also the distance between the starting position of pRNA synthesis and stem-loop I, are essential structural features for G4oric function. PMID- 2558063 TI - Carcinogenicity study of 3,3'-dimethylbenzidine dihydrochloride in BALB/c mice. AB - BALB/c mice (120/sex/dose) were given 0, 5, 9, 18, 35, 70 or 140 ppm of 3,3' dimethylbenzidine dihydrochloride in their drinking-water and killed after 13, 26, 39, 52, 78 or 116 wk. Full histopathological evaluations were performed on all animals that were found dead or moribund, or that were killed on schedule. Fatal lung alveolar cell neoplasms began to appear in males receiving 140 ppm at 78 wk and there was a significant dose-related decrease in the time-to-death from this cause. There were no significant dose-related trends for this neoplasm in females, nor were there treatment-related effects on body weight, water consumption or other lesions in either sex. This is the first report of a neoplastic response to 3,3'-dimethylbenzidine dihydrochloride in mice. PMID- 2558064 TI - Alpha-adrenoceptor properties in rat strains sensitive or resistant to salt induced hypertension. AB - Cerebral and renal alpha 2-adrenoceptors are implicated in the control of sympathetic activity and of sodium reabsorption respectively. In addition, sodium ions play an important role in the regulation of either alpha 2-adrenoceptor densities and affinities for adrenergic agonists. In the present study, alpha adrenoceptor properties were investigated in genetically predetermined salt sensitive and salt-resistant Dahl and Sabra rats. Cerebral alpha 2-adrenoceptor densities were higher in salt-resistant than in salt-sensitive Dahl and Sabra rats. In contrast, renal alpha 2-adrenoceptor density was higher in salt sensitive than in salt-resistant rats. No difference in cerebral and renal alpha 1-adrenoceptor densities was observed between Dahl and Sabra substrains. Noradrenaline content in cerebral and renal cortex were also similar in both these rat substrains. Sodium ions markedly increased cerebral and renal high affinity alpha 2-adrenoceptor densities in salt-sensitive but not in salt resistant rats. Cerebral and renal alpha 1-adrenoceptor densities were unchanged in salt-sensitive and salt-resistant substrains of Dahl and Sabra rats. In addition, sodium ions reduced the affinity of adrenaline for renal alpha 2 adrenoceptors in salt-sensitive rats but not in salt-resistant rats. We can conclude that there exist genetically determined differences in the densities and properties of cerebral and renal alpha 2-adrenoceptors between salt-sensitive and salt-resistant rat strains. Abnormal densities of alpha 2-adrenoceptors may play a primary role in the role in the development of hypertension in salt-sensitive animals. These results also suggest an association between absence of sodium regulation of alpha 2-adrenoceptors and resistance to salt-induced hypertension. The absence of sodium regulation in salt-resistant rats may be linked either to a particular receptor conformation or to an abnormal structure of the receptor system. This property may represent a genetically-mediated change responsible for the resistance to the development of salt-induced hypertension. PMID- 2558065 TI - Expression of desmin and myoglobin in rhabdomyosarcomas and in developing skeletal muscle. AB - Immunohistochemical staining for desmin and myoglobin was investigated in 35 rhabdomyosarcomas from young people and in skeletal muscle from 16 human fetuses of known gestational age. Twenty-nine of the rhabdomyosarcomas expressed desmin but six undifferentiated or poorly-differentiated tumours were desmin negative. Of the desmin positive cases, most undifferentiated or poorly-differentiated sarcomas expressed desmin alone (12/35). Tumours with increasing rhabdomyoblastomic differentiation co-expressed myoglobin (9/35) and well differentiated examples also contained cross-striations (7/35). Skeletal muscle from fetuses aged 8 weeks or less consisted mainly of primitive desmin negative round cells. As the cells began to differentiate they quickly expressed desmin and, at approximately 10 weeks, myoglobin was expressed and cross-striations were seen. The combined results strengthen the view that desmin (within a strictly defined context of round cell tumours in young people) is a reliable marker for rhabdomyoblastic differentiation. Support is also given to the notion that very primitive rhabdomyosarcomas may be desmin-negative, although the difficulties of establishing firm diagnoses for some of these tumours is emphasized. PMID- 2558066 TI - Mucinous carcinoma of the breast: further characterization of its three subtypes. AB - Nineteen cases of mucinous carcinoma of the breast were studied. Twelve tumours were of the pure type, and seven were mixed. All had abundant neutral and acidic mucin, and stained strongly with CAM 5.2. Of the 12 pure mucinous tumours, six were devoid of argyrophilic granules and were S-100 negative, and only one was CEA positive. All six patients are alive with no evidence of recurrence (mean follow-up 42 months). The other six pure mucinous tumours were rich in argyrophilic granules. Five of these showed S-100 positivity and all were CEA positive. One patient developed local recurrence and one died of myocardial infarction with no evidence of tumour recurrence (mean follow-up 80 months). Of the seven mixed tumours, only one contained an occasional cell with argyrophilic granules and four had variable degrees of CEA positivity. Two patients died and one developed bony metastasis (mean follow-up 40 months). Our findings emphasise the microscopic and prognostic differences between the three subtypes of mucinous carcinoma of the breast, and support the concept of dividing pure mucinous tumours into two distinct subtypes. We suggest that the latter subtyping can be qualitatively made on the basis of the presence or absence of argyrophilic granules in the tumour cells. PMID- 2558067 TI - Cytogenetic and molecular analysis of an unbalanced translocation (X;7) (q28;p15) in a dysmorphic girl. AB - A severely retarded and dysmorphic girl, carrying an unbalanced X/7 translocation with breakpoints at Xq28 and 7p14, was analyzed by cytogenetic, biochemical and molecular techniques. The X/7 translocated chromosome was found to replicate consistently late in the 105 metaphases analyzed. In 83 of these cells, late replication was limited to the X portion of the abnormal chromosome, whereas in 22 cells incomplete spreading into the autosomal fragment was observed. Southern blot and in situ hybridization experiments with probe G80 (locus D7S373) (previously localized to 7p13-15) and G98 (localized to 7p14-15) assigns the former to 7p15 and the latter to 7p14, thus suggesting the order 7ter-G80-G98 cen. The activity of the enzyme phosphoserine phosphatase localized to 7pter-p14 was increased. Southern blotting experiments with 19 probes spanning the entire X chromosome demonstrated that the translocated chromosome had lost a portion of Xq28 (locus DXS51) but still retained part of Xq27 (F9 locus). The results confirm that the proband is trisomic for the region 7p15-pter and monosomic for the region Xq28-qter. Comparing her phenotype with those of other cases of partial trisomy or monosomy 7p, we confirm that band 7q21 is probably involved in skull development. PMID- 2558068 TI - Localization of the active type I DNA topoisomerase gene on human chromosome 20q11.2-13.1, and two pseudogenes on chromosomes 1q23-24 and 22q11.2-13.1. AB - Different subfragments of a cDNA coding for DNA topoisomerase I were used as probes to determine the chromosomal localization of topoisomerase I sequences in human cells. Southern blotting of restricted DNA from a panel of rodent-human somatic cell hybrids revealed the localization of the complete gene on chromosome 20 and the presence of two truncated topoisomerase I pseudogene sequences on chromosomes 1 and 22. In situ chromosome hybridization experiments confirmed these results showing the location of the complete gene on band q11.2-13.1 of chromosome 20, and the location of the pseudogene sequences on band q23-24 of chromosome 1 and q11.2-13.1 of chromosome 22. PMID- 2558069 TI - DNA fingerprinting with the oligonucleotide probe (CAC)5/(GTG)5: somatic stability and germline mutations. AB - DNA fingerprints were generated from various human somatic tissues and from peripheral blood of 179 children and their 80 parents using (CAC)5/(GTG)5 oligonucleotide probes. Whereas somatic stability of the fingerprint patterns was demonstrated, the average rate for germline mutations was estimated to be approximately 0.001 per DNA locus and gamete, with the three different restriction enzymes used. Seven out of eight mutations observed appeared to be of paternal origin. PMID- 2558070 TI - Studies on the uptake, binding and metabolism of leukotriene B4 by human neutrophils. AB - Human polymorphonuclear granulocytes (PMN) generate the inflammatory mediator leukotriene B4 (LTB4) as a response to cell activation. In addition, PMN inactivate LTB4 by omega-oxidation resulting in the formation of 20-OH- and 20 COOH-LTB4. The transport of exogenous LTB4 to the metabolizing enzymes is mediated via high- and low-affinity receptor subsets. Uptake of [3H]LTB4 by the cells was carried out in a time-dependent fashion, reaching maximal values after 5 min of incubation. No additional uptake of [3H]LTB4 then occurred. Prestimulation of PMN with phorbol myristate acetate or sodium fluoride resulted in the loss of high- and low-affinity receptors. Deactivating concentrations of LTB4 specifically reduced the high-affinity receptor subset. Prestimulation of PMN with cytochalasin B or with the membrane fluidizer butanol shifted the low affinity receptors to the high-affinity state. The polyene antibiotic amphotericin B shifted high-affinity receptors to the low-affinity subset. The changes in the receptor expression pattern correlated with the respective conversion rate of exogenously added LTB4. Our results suggest that the distribution of high- and low-affinity receptors is regulated by GTP-binding proteins, the activation of protein kinase C and the organization of the membrane bilayer. In this way, human neutrophils control the respective level of the lipid mediator LTB4. PMID- 2558072 TI - Cyclic AMP level of lymphocytes in patients with systemic lupus erythematosus and its relation to disease activity. AB - The basal and stimulated intracellular cyclic AMP (cAMP) levels of peripheral blood mononuclear cells (PBMC) of 16 control subjects and 14 patients with systemic lupus erythematosus (SLE), all fulfilling the ARA criteria, were studied. No significant difference in basal cAMP level was observed between SLE patients and controls. SLE lymphocytes (both active and inactive) elicited a diminished response to aminophylline and prostaglandin E2 (PGE2). No correlation was seen between disease activity and either baseline cAMP levels or response to these stimulators. We suggest an intrinsic (not disease activity-related) impairment of the adenylate cyclase-dependent regulatory mechanism in the PBMC of SLE patients, which may result in a defective IL-2 production and IL-2 dependent biological functions. PMID- 2558073 TI - Influence of prolactin on neural and glial cellular adenosine triphosphatases in immature male bonnet monkeys (Macaca radiata, Geoffroy). AB - The effect of prolactin on specific activities of adenosine triphosphatases (ATPases) in neural and gliar cells of cerebral cortex, cerebellum and pons medulla of immature male bonnet monkeys was studied. Na+, K+ dependent ATPase was stimulated, while Mg2+ and Ca2+ dependent ATPase activities showed reduction in neural as well as glial cells of cerebral cortex and cerebellum. However, in pons medulla, Na+, K+ and Mg2+ dependent ATPases showed the same trend in neural and glial cells, respectively, as in the other two regions. The data obtained reveal that prolactin has specific effect on different ATPases, in different regions of the brain. PMID- 2558074 TI - In vitro effect of lorazepam on Mg2+ and (Na+,K+) ATPases of human foetal brain. AB - Lorazepam (LZ), a benzodiazepine group of drug, inhibits Mg2+ and (Na+,K+) ATPases (EC 3.6.1.3) activity of human foetal and adult brain. The inhibitory effect neither varied with respect to the region (i.e. cerebrum and cerebellum) nor with the age of the foetus. The inhibition of ATPases activity indicates that the neuronal transmission processes, may be affected and raises the possibility of developmental disturbances. PMID- 2558071 TI - Monocyte superoxide secretion triggered by human IgA. AB - While there is much evidence for a key role of IgA in mucosal defence, its mode of action is incompletely understood. The finding of Fc receptors for IgA on various phagocytic cells has led to examination of the ability of IgA to mediate the protective functions of these cells. We studied the ability of human peripheral monocytes to secrete superoxide upon interaction with human IgA, IgG or IgM bound to a solid phase. Both secretory and serum IgA triggered the superoxide response, producing superoxide levels comparable to those induced by IgG, whereas IgM and mouse IgA were inactive. A combination of monomeric IgG and a monoclonal anti-IgG Fc receptor antibody inhibited superoxide secretion mediated through IgG but failed to block the IgA-triggered response, demonstrating that IgA was recognized through specific receptors. In addition IgA was capable of mediating phagocytosis when attached to erythrocytes. PMID- 2558075 TI - Demonstration of histamine receptors on the surface of Entamoeba histolytica. AB - Histamine receptors on the surface of E. histolytica could be demonstrated by histochemical method using three isolates of the protozoa grown and maintained in modified Boeck and Drbohlav's medium. Prior treatment of E. histolytica with cimetidine a H2 blocker, blocked the histamine uptake. Similar treatment with mepyramine maleate, a H1 blocker, did not prevent histamine uptake by the protozoa. It is postulated that E. histolytica has H2 receptors on its surface. PMID- 2558076 TI - Transforming activity of nasopharyngeal carcinoma DNA detectable in mouse JB6 cells. AB - A JB6 mouse epidermal recipient cell line has been used to detect nasopharyngeal carcinoma (NPC) DNA-associated transforming activity that is not detectable in the NIH 3T3 focus assay. NPC DNA showed both transforming activity and activity for transferring sensitivity to tumor-promoter-induced neoplastic transformation, assayed in 2 different variants of mouse JB6 cells. Comparison of DNAs from various NPC sources that did or did not harbor EBV DNA and that varied in degree of differentiation showed similar transforming activities and similar activities for transferring promotion sensitivity. Thus both a NPC DNA-associated promotion sensitivity and an oncogenic activity function independently of concurrent EBV gene expression. PMID- 2558078 TI - Monoclonal antibody UJ13A recognizes the neural cell adhesion molecule (NCAM). AB - Monoclonal antibody (MAb) UJ13A, raised against 16-week-old human foetal brain, recognizes an antigen present on the majority of tissues of neuro-ectodermal origin. The binding profile of this antibody is similar to the known distribution of the neural cell adhesion molecule (NCAM). Although detailed immunohistological and immuno-cytochemical studies with the MAb have been published previously, we demonstrate here, through investigations of selected tissues and cell lines, that the binding profile of an anti-NCAM antiserum is similar to UJ13A. Additional indirect evidence suggesting that UJ13A recognizes NCAM comes from Northern blot analysis of cell lines either binding or not binding UJ13A as determined by indirect immunofluorescence. Only those cell lines known to bind UJ13A express high levels of NCAM mRNA. Western blot analysis of extracts of human brain show that UJ13A recognizes proteins of 180 and 140 kDa in addition to a very weak band of 120 kDa. This data corroborates the suggestion that UJ13A recognizes NCAM as these 3 isoforms of the protein are identified in human brain. Final confirmation of the specificity of UJ13A comes from the study of 3T3 fibroblasts transfected with a cDNA coding for the 125 kDa isoform of human muscle NCAM. UJ13A selectively recognizes these transfectants and binds to a 125 kDa protein isolated from the cells by Western blot analysis. Thus we conclude from these immunological, biochemical and molecular studies that the antigen recognized by the UJ13A MAb is the neural cell adhesion molecule. PMID- 2558077 TI - Antioxidant enzyme activities in normal and transformed mouse liver cells. AB - Copper- and zinc-containing superoxide dismutase (CuZnSOD), manganese-containing superoxide dismutase (MnSOD), catalase (CAT), glutathione peroxidase (GPX, both Se-dependent and Se-independent), and glutathione reductase (GR) were measured in normal, nitrosoguanidine-transformed and SV40-transformed mouse liver cells in culture, as well as in mouse liver homogenates. Enzyme activities were compared on the basis of 3 different endpoints: per mg protein, per mg DNA, and per 10(6) cells. Except for GR, activity of all the measured anti-oxidant enzymes was much higher in vivo than in vitro. All of the anti-oxidant enzyme activities were lower in general in the 2 transformed cell lines than in the in vitro normal cell line, except Cu-ZnSOD, which showed little change. However, MnSOD was the only enzyme which showed lowered activity in both transformed cell lines, no matter what endpoint was used. This finding is in agreement with previous work showing lowered MnSOD activity in tumor cells. PMID- 2558080 TI - Transformation of normal human melanocytes and non-malignant nevus cells by adenovirus 12-SV40 hybrid virus. AB - Infection of normal human melanocyte and nevus cultures with an adenovirus 12 Simian Virus 40 hybrid virus (Ad12-SV40) produced transformed cells that expressed SV40-T antigen. The Ad12-SV40 cells exhibited rapid cell proliferation to high cell densities and efficient growth in soft agar, but none of 15 transformed melanocyte and nevus cultures formed tumors when injected s.c. or under the renal capsule into athymic nude mice. While the Ad12-SV40-transformed cells lost certain properties associated with the melanocytic phenotype, i.e., pigmentation, tyrosinase activity and melanosome content, the expression of melanoma-associated antigens, including nerve growth factor receptor, p97 melano transferrin, and chondroitin sulfate proteoglycan, remained stable. The transformed melanocytes acquired the ability to express HLA-DR antigen, which is found on nevus and melanoma cells. Total ganglioside patterns in Ad12-SV40 transformed cells changed to reflect more advanced stages of tumor progression. Transformed melanocytes, like nevus and melanoma cells, showed increased GD3 content and transformed nevus cells increased GD2 which is a feature of malignant melanoma cells. Ad12-SV40-transformed human melanocytes and nevus cells are useful tools for studying tumor progression under experimental conditions. PMID- 2558081 TI - Cytogenetic analysis of eight human papillomavirus immortalized human keratinocyte cell lines. AB - Cytogenetic analysis was performed on human papillomavirus (HPV)-immortalized cell lines. Lines were established by co-transfection of primary human keratinocyte cells with HPV type 16 or 18 DNA and pSV2neo. The resulting clonal lines contained integrated HPV DNA and exhibited extended life spans in culture but were non-tumorigenic in nude mice. Two HPV16-immortalized lines (FEPE1L8 and FEPE1L9) and 4 HPV18-immortalized lines (FEA, FEH18L, FEP18-5 and FEP18-11) were established. Two additional lines were derived by subsequent treatment of the FEA line with TPA and by further transfection with HSVII DNA. Cytogenetic analysis revealed that all lines were abnormal, containing a variety of numerical and structural aberrations. Six of the 8 lines were hyper-triploid and 2 were near diploid. Examination of lines FEA, FEH18L and FEP18-11 at multiple passages in culture revealed that the lines were clonal and chromosomally stable over extended passage in culture. Structural rearrangements were most common in chromosomes 1 and 3 but also occurred in chromosomes 5, 7, 8, 12, 16 and 22. Marker chromosomes were present in all cell lines. A small metacentric marker, possibly an isochromosome for the short arm of chromosome 5, was consistently present in the FEA line and its derivatives (FEAB10 and FEAT) as well as the FEH18L line. A loss or reduction in copy number of chromosome 13 was seen in 5 of the 8 cell lines. PMID- 2558079 TI - Volatile nitrosamine levels and genotoxicity of food samples from high-risk areas for nasopharyngeal carcinoma before and after nitrosation. AB - Traditional life-style, especially food habits, infection by Epstein-Barr virus (EBV) and genetic factors, have been associated with an increased risk of nasopharyngeal carcinoma (NPC). N-Nitroso compounds and other carcinogens either present in food or formed endogenously, as well as food constituents that activate EBV, have been suspected as etiological factors in NPC pathogenesis. For their characterization preserved food items, frequently consumed in NPC endemic areas in Tunisia, South China and Greenland, were sampled and screened for the presence of mutagens and volatile nitrosamines before and after nitrosation. Aqueous extracts as well as 2 organic extracts of the samples were assayed for genotoxicity in 2 Salmonella typhimurium strains and the SOS chromotest. The same extracts had previously been analyzed for volatile nitrosamines and for EBV activating substances in Raji cells. In our study, 13 out of 16 food samples showed a weak, directly-acting genotoxicity in the SOS chromotest in at least one of the extracts, but only one sample from Greenland was found to be weakly mutagenic in Salmonella TA 98. Chemical nitrosation for 9 out of 15 samples of aqueous food extracts increased the genotoxic effect in the SOS chromotest. Levels of volatile nitrosamines were also elevated for 12 out of 15 samples; highest levels of N-nitrosodimethylamine were found in hard salted and dried fish from China (1,200 micrograms/kg) and highest N-nitrosopyrrolidine levels in a Tunisian spice (3,840 micrograms/kg). In non-nitrosated aqueous food extracts, the level of volatile nitrosamines and genotoxic activities were not correlated with the EBV-inducing activity of the same samples. After chemical nitrosation, EBV-inducing activity was decreased or showed no change and was not correlated with increases in either the genotoxicity or the nitrosamine levels. Our results suggest that EBV-activating compounds belong to a different class of substances. However, there was an association between the changes in genotoxicity and nitrosamine levels due to nitrosation. PMID- 2558083 TI - Metastatic mixed mullerian tumour of the esophagus. AB - Involvement of the esophagus by metastatic disease from malignancies of other distant organs is uncommon. A case of malignant mixed mullerian tumour metastatic to the esophagus is reported. Though a variety of malignancies is known to metastasize to the esophagus, metastasis from malignant mixed mullerian tumour has not been reported, as yet. PMID- 2558084 TI - Carbon-13 magnetic resonance spectroscopy. Problems and promise. AB - Carbon-13 magnetic resonance spectroscopy (13C MRS) is being used to investigate metabolism in the brain, liver, heart, and skeletal muscle of animals. The tricarboxylic acid cycle, glycolysis, gluconeogenesis, hepatic ketogenesis, and metabolism of ethanol are some of the metabolic processes amenable to 13C MRS. Clinical investigation using 13C in humans will become widespread as improvements in sensitivity, localization, enrichment, and decoupling are realized. Carbon-13 MRS offers potential as a new investigative probe into organ physiology. PMID- 2558082 TI - In vivo investigation of myocardial perfusion, metabolism and receptors by positron emission tomography. AB - Positron emission tomography (PET) is a safe noninvasive visualization technique which gives an accurate and quantitative representation of the spatial distribution of a positron-emitting radionuclide in any desired transverse section of the body. Short-lived positron emitters such as Carbon-11 (half life:20 min.). Nitrogen-13 (10 min), Fluorine-18 (110 min) or Bromine 75 can be incorporated into biological molecules:amino acids, sugars, fatty acids, hormones and receptor agonists or antagonists. The PET approach may be compared to quantitative autoradiography with the added advantage of allowing in vivo kinetic studies in man, under normal and pathological conditions. Regional myocardial perfusion is measured with rubidium-82, 13N-labelled ammonia, H2(15) O or albumin microspheres labelled with Gallium-68. Regional myocardial metabolism is assessed with 11C-palmitate and 18F-fluorodeoxyglucose. A high and spatially homogenous accumulation of 11C-palmitate is seen in normal myocardium. PET delineates an increased regional accumulation of 18F-2-FDG in ischemic areas while 11C palmitate accumulation is markedly reduced. Positron Emission Tomography offers a unique opportunity to study receptors in vivo in normal and disease conditions. A potent muscarinic antagonist, 11C-MQNB is used to study the myocardial acetylcholine receptor and beta-blockers, 11C-Pindolol and 11C-CGP 12177 are used to study the beta-adrenergic receptor. The peripheral-type benzodiazepine receptor has been characterized with 11C-PK 11195. PMID- 2558085 TI - Shift-reagent-aided 23Na NMR spectroscopy. AB - Nondestructive observation of intracellular sodium (Na+i) levels is of utmost clinical and biochemical importance, 23Na is a relatively sensitive nuclear magnetic resonance (NMR)-observable nuclide, and NMR is intrinsically noninvasive. Since, however, the frequency position of Nai+ and Nao+ is identical, paramagnetic dysprosium complexes have to be used as shift reagents. The latter differentiate between signals from intracellular and extracellular spaces, providing a nondestructive, continuous method to monitor intracellular cation concentration, in real time. In this article, the methodology of shift reagent-aided 23Na NMR spectroscopy is discussed. Application is demonstrated by results from two studies in isolated perfused rat hearts. In hearts subjected to hypoxia (or ischemia) followed by reoxygenation (or reflow), the authors show that recovery following an ischemic or hypoxic insult can be predicted by monitoring Na+i levels by NMR. In a study in paced hearts, the authors show an accumulation of Na+i with increased heart rate, and also a positive coupling between elevated sodium levels and increased systolic pressure during the pacing up period, as predicted by the Na-pumplag theory. PMID- 2558086 TI - Clinical MRS studies of the brain. AB - Image-guided 31P and 1H magnetic resonance localized spectroscopy was performed on patients with brain tumors, temporal lobe epilepsy, chronic brain stroke, and deep white matter lesions. Absolute molar concentrations of metabolites, peak area ratios, and pH were obtained. The important findings were that 31P metabolite concentrations were significantly reduced in tumors, infarcts, and deep white matter lesions. Similarly, 1H metabolite intensities were reduced in chronic stroke. In the seizure foci of epilepsy patients, in tumors, and in chronic stroke, the pH was more alkaline than the normal pH. Peak area ratios were altered in tumors (reduction of phosphocreatine/inorganic phosphate (PCr/Pi) and in chronic stroke (large increases in Cr/NAA and Cho/NAA). Finally, the spectroscopic imaging technique offers a versatile alternative to the "single point" techniques, producing spectra or images of the spatial distribution of individual 31P metabolites. PMID- 2558087 TI - Separation of lipids from lactate in experimental allergic encephalomyelitis by zero quantum NMR techniques. AB - Using localized proton spectroscopy, the author and associates previously have observed an increase in the resonance at 1.2 +/- 0.2 ppm in the brain of monkeys with experimental allergic encephalomyelitis (EAE). In proton nuclear magnetic resonance (NMR) spectroscopy, the lactate methyl protons resonate at dose to the same chemical shift frequency as the lipid methylene protons (1.2 +/- 0.2 ppm). Noninvasive zero quantum NMR techniques were used to separate lipids from lactate in the brain during the course of EAE development. Out of 53 zero quantum NMR measurements (from three animals), 16 measurements were made at a time when a resonance at 1.2 ppm appeared (after the animals developed EAE). In all these cases, lactate was not detectable. The fact that the resonance (identified as lipids with zero quantum techniques) correlated with histochemical Oil red O staining suggests that these mobile NMR signals are associated with demyelination. PMID- 2558088 TI - NMR spectroscopic assessment of altered myocardial lipid metabolism. AB - Proton nuclear magnetic resonance (NMR) spectroscopy of myocardial lipids can be used to monitor the metabolic activity of the heart. Alterations in lipid NMR signals occur with disorders such as myocardial ischemic events and postischemic dysfunction ("stunned myocardium"). Proton NMR-derived information may prove useful in evaluating the extent and stage of the ischemic injury and viability of the myocardium. PMID- 2558089 TI - Tumor characterization using unconventional MR modalities. AB - This article reviews many of the promising new magnetic resonance (MR) modalities for the characterization of tumor structure and metabolism, excluding 31P magnetic resonance spectroscopy (MRS). The report focuses on recent work with localized proton MRS, MR imaging of diffusion and perfusion, and work on imaging bound pools of sodium ions. The technical problems associated with these methods are discussed, along with the unique information each provides. Many of these techniques have already been used in studies of human disease, or are of sufficient importance that clinical applications are imminent. PMID- 2558090 TI - Antibodies to Epstein-Barr viral antigens in familial rheumatoid arthritis. AB - Serum antibodies to Epstein-Barr virus (EBV) were measured in members of twenty nine families in which two or more first degree relatives had rheumatoid arthritis (RA). These patients were assessed clinically for activity of disease and their treatment recorded. A higher proportion of seropositive individuals was found among patients and affected kindred than among the non-affected groups. Antibody titres to Viral Capsid Antigen (VCA) tended to be higher in patients and affected kindred. In those individuals in whom antibody to Early Antigen (EA) was present there was a lower titre of antibody to Epstein-Barr Nuclear Antigen (ENBA). The results suggest that an immunoregulatory defect may exist in some patients with RA which allows for enhanced expression of EBV. PMID- 2558091 TI - In vitro effects of an acyltripeptide, FK565, on antitumor effector activities and on metabolic activities of human monocytes and granulocytes. AB - In vitro effects of an immunostimulatory acyltripeptide, FK565, on antitumor and metabolic activities of human leukocytes were studied. Monocyte cytotoxicity against A375 melanoma targets was significantly increased following pretreatment with FK565 at concentrations of 1 microgram/ml or more. The tripeptide also up regulated anti-tumor cytostasis by monocytes and showed a strong stimulatory effect on superoxide generation by resting monocytes over a wide range of FK565 concentrations after 18 h preincubation. The monocyte preparations contained an average of 76% LeuM3+HLA-DR+, 12% LeuM3+HLA-DR- and 10% LeuM3-HLA-DR+ cells, and this phenotype distribution was not altered after incubation with FK565. At concentrations above 1 microgram/ml and after 2 h preincubation, FK565 also increased superoxide generation by resting but not stimulated granulocytes. Pre exposure of cultured bovine endothelial cells to the peptide resulted in a significant inhibition of fMLP-stimulated granulocyte adherence to these cells. These data indicate that in vitro incubation of human monocytes and granulocytes with FK565 (0.1-100 micrograms/ml) had resulted in simultaneous up-regulation of several anti-tumor functions mediated by these cells. PMID- 2558092 TI - [A new concept of the etiopathogenesis and prevention of atopic dermatitis]. AB - The hypothesis proposed for the pathogenesis of atopy links the well-known alterations in cell-mediated and humoral immunity, the disturbances of mediator metabolism and the increased disposition of atopic epidermis for inflammation to a common underlying deficiency in the production of prostaglandin E1. The PGE1 deficiency is explained as the result of reduced delta-6-desaturase activity in atopic patients. The development of depressed cell-mediated immunity is regarded as a PGE1-dependent T-cell-maturation defect of the newborn's immune system post partum. Our hypothesis offers a novel approach to the prevention of atopy by administration of gamma-linolenic acid to newborns with increased risk of atopy and to nursing atopic mothers. Furthermore, it provides an explanation for the beneficial therapeutic effects of dietary supplementation of gamma-linolenic acid in patients with atopic dermatitis. PMID- 2558093 TI - Factors affecting the placental transfer of actinides. AB - The primary goal of this paper is to consider factors that affect the availability and transport of actinides from maternal blood, through the placenta, to the conceptus. These factors, of particular importance in scaling results from animals to man, include the route and temporal pattern of administration, the mass and physicochemical state of material administered, metabolism of the pregnant animal and fetal organs or tissue, and species specific changes in placental structure relative to stage of gestation at exposure. Preliminary concepts for descriptive and kinetic models are proposed to integrate these results, to identify additional information required for developing more comprehensive models, and to provide a basis for scaling to human pregnancies for purposes of radiation dosimetry. PMID- 2558094 TI - The use of animal experiments for assessing annual limits on intake and interpreting chest-monitoring data for workers exposed to industrial actinide bearing dusts. AB - The metabolic behavior of 239Pu and 241Am present in three industrial dusts has been studied after their inhalation by the rat. A comparative experiment has also been carried out with a mixture of these actinides, inhaled as their nitrates. The aim of this work was to provide an experimental basis for assessing limits on intake and to establish whether the 239Pu content in the lungs could be interpolated from measurements of 241Am. The results (1) demonstrate the wide differences in the lung retention kinetics of the actinides and in the absolute and relative amounts which translocate to the blood that can occur for industrially produced materials; (2) show that the annual limits on intake (ALI) for the different materials vary between those postulated for class W and Y compounds by the International Commission on Radiological Protection; (3) indicate that, depending on the nature of the dust, acute intakes of 239Pu equivalent to the ALI can be estimated from 241Am chest-monitoring data at times from a few days up to about 3 y after exposure. PMID- 2558095 TI - The current approach of the ICRP Task Group for modeling doses to respiratory tract tissues. AB - For radiation protection purposes, the International Commission on Radiological Protection (ICRP) Task Group proposes to apportion radiation risk within the respiratory tract according to the tumor mortality rates observed in the different anatomical regions. This approach requires that doses absorbed by extrathoracic tissues must be considered, in addition to those in the lung. For the extrathoracic region, the tissues at highest potential risk are the pharyngeal parts of the nasopharynx and oropharynx and a part of the larynx. In the lung, all tissues are potentially at risk, and it is necessary to consider doses absorbed by bronchial tissues, the lung parenchyma, and lymph nodes. This paper outlines the methods proposed by the Task Group to evaluate the heterogeneous doses absorbed by sensitive cells in these tissues from radioactive decays of alpha-emitters. The objective is to evaluate doses to broad regions of the respiratory tract, where the regions are defined to reflect substantial differences in potential risk when taking into account deposition and clearance behavior. The Task Group proposes to represent the respiratory tract by three generic regions: an extrathoracic region and two thoracic regions, one clearing fast and one slowly. The models of aerosol deposition and clearance applied for each region are outlined. To illustrate the use of the model, doses are evaluated for the key cases of short-lived radionuclides and long-lived insoluble alpha emitters and are discussed with regard to current ICRP recommendations. PMID- 2558096 TI - Purposeful alteration of [99mTc]-pertechnetate biodistribution. AB - A series of imaging studies were conducted in rats to assess the effect of stannous containing chemical species on the normal biodistribution of [99mTc]pertechnetate. The goal of the study was to determine if tissue activity could be altered by use of selected chemical agents and if such alteration could be used to clear non-target activity for enhanced image interpretation and/or to visualize two or more organ systems following a single injection of radioactivity. Two distinct patterns of tissue activity alteration could be induced. Tissue distribution studies in rats demonstrated statistically significant differences between tissue radioactivity in groups selectively studied for the type and magnitude of the induced alteration. PMID- 2558097 TI - Muscle fibre size and number following immobilisation atrophy. AB - Muscle fibre number and cross sectional area were studied in the response to immobilisation atrophy of the long head of the triceps brachii. Following eight weeks of immobilisation, fibre number of the muscle from the immobilised limb was compared to that of the contralateral control limb in six rats. Mean fibre cross sectional area of the LHT from the immobilised limb was compared to that of the contralateral control for another six animals. Atrophy, as estimated by a decrease in wet muscle weight, was 38.0% for the group used for fibre number estimations and 45.7% for the group used for fibre area. Fibre counts revealed no difference between muscles from immobilised and control limbs. Mean fibre area was 42.1% less for the muscle from the immobilised limb compared to the control limb. The results of this study indicate that atrophy of the LHT produced by immobilisation of the forelimb is the result of atrophy of the muscle fibres without a decrease in muscle fibre number. PMID- 2558098 TI - Marchi-positive myelinoid bodies at the transition between the central and the peripheral nervous system in some vertebrates. AB - The CNS-PNS (central nervous system-peripheral nervous system) transitional region of cranial and spinal nerve roots in some vertebrate species was analysed with respect to the occurrence and the distribution of myelinoid Marchi-positive bodies. Both cranial and spinal nerve roots contained more Marchi-positive bodies in their CNS than in their PNS segments. An accumulation of Marchi-positive bodies was usually noted just central to the CNS-PNS borderline. Comparisons between calibre spectra and Marchi index in the cat revealed a particularly high number of Marchi-positive bodies in nerve roots with a high content of myelinated fibres with diameters greater than or equal to 5 microns. Marchi-positive bodies were absent in CNS tissue lacking myelinated nerve fibres. CNS borderline internodes measuring between 200 and 300 microns in length were noted in fibres as thick as 15 microns in feline S1 ventral and dorsal roots. The general picture was similar in all analysed species. Noteworthy however, was the small difference in number of Marchi-positive bodies between CNS and PNS tissue in Xenopus. The chicken contained many myelinoid bodies of similar size and texture as the Marchi positive bodies but without the Marchi-positive staining properties. The results show that normally occurring Marchi-positive bodies in the CNS are more numerous along paranodal segments than along mid-internodal segments of myelinated nerve fibres and thus support the hypothesis that Marchi-positive bodies are preferentially derived from paranodal myelin. PMID- 2558100 TI - Extracorporeal bicarbonate space after bicarbonate or a bicarbonate-carbonate mixture in acidotic dogs. AB - The effects of sodium bicarbonate and a bicarbonate-carbonate mixture on expired CO2 and the volume of distribution of bicarbonate were studied in eight anesthetized, paralyzed, and ventilated dogs made acidotic with HCl (5 mmol/kg) infused over 90 min. Both sodium bicarbonate and Carbicarb resulted in systemic alkalinization and comparable increases in the serum bicarbonate at 50 min (7.07 +/- 0.91 vs. 7.99 +/- 0.77, respectively; P = NS). Sodium bicarbonate infusion resulted in an increase in CO2 excretion that accounted for a fractional CO2 excretion of 0.20 +/- 0.09, whereas infusion of a bicarbonate-carbonate mixture resulted in a fractional CO2 excretion of -0.06 +/- 0.09 (P less than 0.01). The uncorrected volume of distribution of bicarbonate after sodium bicarbonate infusion was higher than that seen with the bicarbonate-carbonate mixture (0.60 +/- 0.07 vs. 0.34 +/- 0.03 l/kg; P less than 0.01). However, when the volume of bicarbonate distribution was corrected for expired CO2, there was no difference between treatment with sodium bicarbonate and the bicarbonate-carbonate mixture (0.44 +/- 0.07 vs. 0.38 +/- 0.04 l/kg; P = NS). These data demonstrate that, in this animal model of acidosis, sodium bicarbonate treatment of systemic acidosis is accompanied by a generation of a considerable amount of CO2, whereas treatment with a bicarbonate-carbonate mixture is not. This suggests that in states of impaired ventilation, a bicarbonate-carbonate mixture may offer more efficient systemic alkalinization and may be associated with less CO2 generation than sodium bicarbonate. PMID- 2558101 TI - Zymosan-activated plasma causes prolonged decreases in PMN superoxide release in sheep. AB - It is well established that activation of neutrophils within the pulmonary circulation produces acute lung injury in which adherence of neutrophils to endothelial cells is an obligatory step in the mechanism of injury. The effects of in vivo activation of neutrophils on the in vitro responses of these cells to stimulation have not been determined, although such information may be important in understanding how different etiological factors may interact to produce infection or acute respiratory failure. By using an assay to sequentially measure superoxide anion (O2-) release from adherent neutrophils stimulated with phorbol myristate acetate (PMA), we measured the in vitro activation response of peripheral blood neutrophils isolated before and 24 h after infusion of zymosan activated plasma (ZAP; or untreated plasma as a control), air bubbles, or PMA in awake, instrumented sheep. Each of the three inflammatory agents produced an increase in lung microvascular permeability characteristic of acute lung injury; control plasma did not. For the in vivo ZAP experiments, stimulated O2- release in vitro by using PMA was approximately 50% lower (P less than 0.05) for neutrophils isolated 24 h after the in vivo infusion (4.3 +/- 0.8 nmol/500,000 cells) than before (8.1 +/- 0.2 nmol/500,000 cells). For the air emboli or PMA in vivo experiments, there were no changes in neutrophil activation responses in vitro. Similarly, infusion of control plasma did not result in reduced neutrophil O2- release. These results show that alterations in the inflammatory potential of neutrophils may occur in vivo and that such alterations appear to be dependent on the mechanism and agent by which lung injury is produced. PMID- 2558102 TI - Degradative enzymes modulate airway responses to intravenous neurokinins A and B. AB - We studied the effects of the neutral endopeptidase (NEP) inhibitor thiorphan (1.7 mg/kg iv) and the angiotensin-converting enzyme (ACE) inhibitor captopril (5.7 mg/kg iv) on airway responses to rapid intravenous infusions of neurokinin A (NKA) and neurokinin B (NKB) in anesthetized, mechanically ventilated guinea pigs. The dose of NKA required to decrease pulmonary conductance to 50% of its base-line value (ED50GL) was fivefold less (P less than 0.0001) in animals treated with thiorphan compared with controls. NKA1-8, a product resulting from cleavage of NKA by NEP, had no bronchoconstrictor activity. Similar results were obtained by using NKB as the bronchoconstricting agent. Captopril had no significant effect on airway responses to NKA or NKB. In contrast, both thiorphan and captopril decrease the ED50GL for substance P (SP). We also compared the relative bronchoconstrictor potency of NKA, NKB, and SP. In control animals, the rank order of ED50GL values was NKA much less than NKB = SP. NKA also caused a more prolonged bronchoconstriction than SP or NKB. Thiorphan had no effect on the rank order of bronchoconstrictor potency, but in animals treated with captopril, the rank order of ED50GL values was altered to NKA less than SP less than NKB. These results suggest that degradation of NKA and NKB by NEP but not by ACE is an important determinant of the bronchoconstriction induced by these peptides. The degradation by ACE of SP but not NKA or NKB influences the observed relative potency of the three tachykinins as bronchoactive agents. PMID- 2558099 TI - Muscle malonyl-CoA decreases during exercise. AB - Malonyl-CoA, the inhibitor of carnitine acyltransferase I, is an important regulator of fatty acid oxidation and ketogenesis in the liver. Muscle carnitine acyltransferase I has previously been reported to be more sensitive to malonyl CoA inhibition than is liver carnitine acyltransferase I. Fluctuations in malonyl CoA concentration may therefore be important in regulating the rate of fatty acid oxidation in muscle during exercise. Male rats were anesthetized (pentobarbital via venous catheters) at rest or after 30 min of treadmill exercise (21 m/min, 15% grade). The gastrocnemius/plantaris muscles were frozen at liquid N2 temperature. Muscle malonyl-CoA decreased from 1.66 +/- 0.17 to 0.60 +/- 0.05 nmol/g during the exercise. This change was accompanied by a 31% increase in cAMP in the muscle. The decline in malonyl-CoA occurred before muscle glycogen depletion and before onset of hypoglycemia. Plasma catecholamines, corticosterone, and free fatty acids were all significantly increased during the exercise. This exercise-induced decrease in malonyl-CoA may be important for allowing the increase in muscle fatty acid oxidation during exercise. PMID- 2558103 TI - Dibutyryl cAMP, aminophylline, and beta-adrenergic agonists protect against pulmonary edema caused by phosgene. AB - Phosgene is a toxic oxidant gas that causes the adult respiratory distress syndrome in exposed workers. Phosgene exposure markedly increased lung weight gain in buffer-perfused isolated rabbit lungs (31 +/- 5 g over 60 min after phosgene vs. 7.7 +/- 1.2 in control lungs, P less than 0.01) and markedly increased the lung leak index for 125I-albumin (0.28 +/- 0.03 after phosgene vs. 0.02 +/- 0.01 in control lungs, P less than 0.01). Pretreatment with dibutyryl adenosine 3',5' -cyclic monophosphate (DBcAMP), aminophylline, or terbutaline plus isoproterenol prevented the increase in lung weight caused by phosgene (31 +/- 5 g phosgene, 11.7 +/- 2.8 DBcAMP, 7.5 +/- 2.5 aminophylline, 6.1 +/- 1 terbutaline and isoproterenol, 6.1 +/- 1.2 control + aminophylline, and 7.7 +/- 1.2 control; all treatments were P less than 0.01 vs. the untreated phosgene group and not significantly different from control lungs). Pretreatment with aminophylline prevented the increase in lung leak index for 125I-albumin (0.28 +/ 0.03 after phosgene vs. 0.06 +/- 0.02 in aminophylline-treated lungs, P less than 0.01). Posttreatment with aminophylline and terbutaline also prevented the increase in lung weight caused by phosgene. These results indicate that phosgene dramatically increases the movement of fluid and protein across the pulmonary vasculature and that treatment with DBcAMP, aminophylline, terbutaline, or isoproterenol markedly reduces the pulmonary edema caused by phosgene. PMID- 2558104 TI - Prognostic factors in breast cancer. AB - A variety of tumor characteristics can provide prognostic information useful in managing the patient with primary breast cancer. Some of these characteristics are firmly established, whereas others are observer dependent or require prospective validation. No single characteristic, however, is likely to fully define which patient with primary breast cancer is destined to relapse. This clinical dilemma--recognition of the high-risk patient--is particularly important in the management of women with node-negative breast cancer. Because most women with this early stage of disease will be cured by surgery alone, the use of adjuvant chemotherapy must be limited to high-risk subsets. Tumor size and ER status are established prognostic factors. Histologic and nuclear grade may be important, but problems of interobserver variability remain. Some studies have shown that aneuploidy or a high S-phase fraction may be independent, high-risk characteristics. Flow cytometric DNA content analysis must be applied with caution, however, because the calculation of S-phase fraction has not been standardized and because the prognostic utility of this approach has not been prospectively confirmed. For now, a prudent approach might be to gather as much prognostic information about each patient's tumor as possible. Those with several of the high-risk characteristics listed in Table 2 should receive strongest consideration for adjuvant treatment. Some of these same prognostic factors, along with several others, can be used to characterize the high-risk node positive patient. The number of involved axillary nodes is the most important established predictor. Progesterone-receptor status is associated with both disease-free and overall survival, whereas ER status is independently related only to overall survival. Histologic grade, DNA ploidy, and S-phase fraction can also be used to help define the high-risk patient. Finally, tumors that amplify or overexpress the HER-2 gene may have a higher risk of relapse, although this finding has been questioned. Management of patients with breast cancer requires an individualized approach that is based on a careful weighing of a variety of prognostic considerations. The relative importance of these factors will require further large-scale, prospective, multiparameter studies. Although results from such studies are awaited, an understanding of the clinical heterogeneity of breast cancer must be based on a multiplicity of observations, each of which characterizes, in a limited way, the biology of this disease. PMID- 2558105 TI - Therapy of in situ cancer. AB - In situ carcinoma of the breast is divided into ductal carcinoma in situ (DCIS) and lobular carcinoma in situ (LCIS). These two entities have different morphologic appearances and natural histories. The traditional treatment of DCIS has been mastectomy, which has been shown to yield a very high cure rate. Other options include excision with or without radiotherapy. Preliminary evidence suggests that these techniques can be successful in the great majority of patients, with most treatment failures being manageable by salvage mastectomy. Careful observation of these patients appears associated with a low risk of breast cancer mortality and is generally the recommended policy. PMID- 2558107 TI - N-terminal half of a mitochondrial presequence peptide takes a helical conformation when bound to dodecylphosphocholine micelles: a proton nuclear magnetic resonance study. AB - Two-dimensional proton nuclear magnetic resonance (NMR) spectra of a synthetic peptide (p25) corresponding to the amino-terminus of the yeast mitochondrial cytochrome oxidase subunit IV precursor protein have been analyzed. Sequence specific resonance assignments of the peptide have been made in the presence of micelles of a phospholipid analog, perdeuterated dodecylphosphocholine (DPC), with the aid of such techniques as HOHAHA, DQF-COSY, and NOESY. The interresidue nuclear Overhauser effects (NOEs) indicate that the N-terminal half of p25 (S3 F11) takes a helical structure while the C-terminal half does not take a regular secondary structure. Addition of DPC to the solution of p25 induced chemical shift changes only of the resonances from the residues in the N-terminal half, suggesting that the N-terminal half of p25 is directly involved in binding to DPC. The induced helical structure in the N-terminal half at a lipid-water interface may be important in the ability of this presequence to direct a "passenger" protein into mitochondria. PMID- 2558108 TI - Spectrophotometric, electron paramagnetic resonance and oxygen binding studies on the hemoglobin from the marine polychaete Perinereis aibuhitensis (Grube): comparative physiology of hemoglobin. AB - The physicochemical properties of giant hemoglobin (Hb) of the marine polychaete Perinereis aibuhitensis were extensively studied and the following results were obtained. (1) Light absorption spectra of the oxy, deoxy, CO, met, and cyanomet derivatives were similar to those for human Hb, except for a somewhat peculiar shape and pH-dependence of the met derivative, and high absorbance values around 277 nm for all these derivatives of Perinereis Hb. Abnormal pH dependence for the met derivative was confirmed by powder electron parmagnetic resonance (EPR) spectroscopy, which revealed that a water molecule does not coordinate to the heme iron as a sixth ligand. The high absorption around 277 nm is indicative of the existence of some non-heme polypeptide chains and/or a high content of aromatic residues in the molecule. (2) UV difference and derivative spectra revealed oxygenation-induced conformational changes in the protein moiety that are related to the degree of cooperativity. (3) The EPR spectrum for the nitrosyl derivative showed well-resolved triplet-triplet splittings due to 14N, indicating that the proximal residue is probably a histidine. (4) The oxygen affinity and cooperativity of this Hb were pH-dependent. Mg2+ markedly increased the oxygen affinity, the Bohr effect, and the cooperativity, which was maximal at physiological pH. CO2 and anions such as 2,3-diphosphoglycerate and inositol hexaphosphate had no effect on the oxygenation properties. Thus, different from vertebrate Hb, the oxygen-binding properties of this Hb are regulated by divalent cations which bind preferentially to the oxy form. The low temperature-dependence of oxygen affinity observed for this Hb is a sign of adaptation to the environment by this poikilothermic organism. (5) By using a graphic method, the minimal functional unit that preserves the full cooperativity (allosteric unit) was inferred to be the one containing 6 heme groups and its significance is discussed in connection with the structural hierarchy of the molecule. PMID- 2558106 TI - Protein factors which regulate cell motility. AB - Cell motility (i.e., movement) is an essential component of normal development, inflammation, tissue repair, angiogenesis, and tumor invasion. Various molecules can affect the motility and positioning of mammalian cells, including peptide growth factors, (e.g., EGF, PDGF, TGF-beta), substrate-adhesion molecules (e.g., fibronectin, laminin), cell adhesion molecules (CAMs), and metalloproteinases. Recent studies have demonstrated a group of motility-stimulating proteins which do not appear to fit into any of the above categories. Examples include: 1) scatter factor (SF), a mesenchymal cell-derived protein which causes contiguous sheets of epithelium to separate into individual cells and stimulates the migration of epithelial as well as vascular endothelial cells; 2) autocrine motility factor (AMF), a tumor cell-derived protein which stimulates migration of the producer cells; and 3) migration-stimulating factor (MSF), a protein produced by fetal and cancer patient fibroblasts which stimulates penetration of three dimensional collagen gels by non-producing adult fibroblasts. SF, AMF, and MSF are soluble and heat labile proteins with Mr of 77, 55, and 70 kd by SDS-PAGE, respectively, and may be members of a new class of cell-specific regulators of motility. Their physiologic functions have not been established, but available data suggest that they may be involved in fetal development and/or tissue repair. PMID- 2558109 TI - Purification of human lung angiotensin-converting enzyme by high-performance liquid chromatography: properties and N-terminal amino acid sequence. AB - Angiotensin-converting enzyme from the human lung was purified to apparent homogeneity, using high-performance liquid chromatography following trypsin treatment of the detergent-extract. A 1,750-fold purification was achieved with a 26% yield. The specific activity of the enzyme was 105 units per mg protein with the substrate hippuryl-L-histidyl-L-leucine (HHL) at 37 degrees C, and the Km value for HHL was 1.9 mM. The molecular weight was estimated to be 170,000 by sodium dodecyl sulfate gel electrophoresis, and the isoelectric point was about 4.8, by chromatofocusing. The N-terminal amino acid sequence was (NH2)-X-X-Pro Gly-Leu-Glu-Pro-Gly-X-Phe-Ser-Ala-Arg-Glu-Ala-Gly-Ala. This is highly homologous to the corresponding sequences of the enzymes from bovine and rabbit lung and from pig, bovine, and mouse kidney, but significantly different from that of the human kidney enzyme. PMID- 2558110 TI - Molecular cloning and nucleotide sequence of a cDNA encoding Euglena gracilis cytochrome c1. AB - The amino acid sequence of the mature protein of Euglena gracilis cytochrome c1 was determined by sequencing of its cDNA. A cDNA expression library was constructed from Euglena poly(A)+ RNA in phage lambda gt11 and screened with an antiserum raised against cytochrome c1 polypeptide isolated from purified E. gracilis complex III. An isolated cDNA clone consisted of 872 base pairs and encoded the mature protein with 243 amino acids. The deduced amino acid sequence contained the unusual heme binding sequence-Phe-Ala-Pro-Cys-His- (Mukai, K. et al. (1989) Eur. J. Biochem. 178, 649-656) instead of the typical sequence,-Cys-X Y-Cys-His-, commonly found in C-type cytochromes. Comparison of the sequence with those of several other cytochromes c1 revealed that Euglena cytochrome c1 conserved the residues probably ligating heme-iron, those supposed to interact with cytochrome c and regions anchoring the mitochondrial inner membrane. PMID- 2558111 TI - UDP-glucose pyrophosphorylase from potato tuber: purification and characterization. AB - UDP-glucose pyrophosphorylase from potato tuber was purified 243-fold to a nearly homogeneous state with a recovery of 30%. The purified enzyme utilized UDP glucose, but not ADP-glucose, as the substrate, and was not activated by 3 phosphoglyceric acid. Product inhibition studies revealed the sequential binding of UDP-glucose and MgPPi and the sequential release of glucose-1-phosphate and MgUTP, in this order. Analyses of the effects of Mg2+ on the enzyme activity suggest that the MgPPi and MgUTP complexes are the actual substrates for the enzyme reaction, and that free UTP acts as an inhibitor. The enzyme exists probably as the monomer of an approximately 50-kDa polypeptide with a blocked amino terminus. For structural comparison, 29 peptides isolated from a tryptic digest of the S-carboxymethylated enzyme were sequenced. The results show that the potato tuber enzyme is homologous to UDP-glucose pyrophosphorylase from slime mold, but not to ADP-glucose pyrophosphorylase from Escherichia coli, and provide structural evidence that UDP-glucose and ADP-glucose pyrophosphorylase are two different protein entities. PMID- 2558112 TI - Existence of Mg2+-dependent, neutral sphingomyelinase in nuclei of rat ascites hepatoma cells. AB - A sphingomyelinase, which specifically hydrolyzes sphingomyelin into ceramide and phosphocholine, was solubilized from nuclear matrix fraction of rat ascites hepatoma, AH7974 cells. The solubilized enzyme was subjected to Mono Q column chromatography in an FPLC system. The sphingomyelinase which was adsorbed on the column and eluted at 0.25-0.5 M NaCl was characterized. The enzyme required 10 mM MgCl2, 0.01% Triton X-100, 1 mM dithiothreitol, and a higher concentration of buffer than 1 M for its maximal activity, and the optimal pH was 6.7-7.2 in 2 M Tris/acetic acid or 7.5 in 2 M potassium acetate/acetic acid. N-Ethylmaleimide completely inhibited the enzyme activity at 0.2 mM. Therefore, this enzyme is classified as a Mg2+-dependent, neutral sphingomyelinase. The sphingomyelinase sedimented at 4.3S through a 10-30% glycerol gradient containing 2 M potassium acetate. This enzyme was highly specific to sphingomyelin and did not hydrolyze phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylinositol. Various characteristics of the nuclear sphingomyelinase were similar to those of the plasma membrane enzyme except its requirement for a high concentration of buffer and SH-reagent. PMID- 2558114 TI - Purification and characterization of membrane-bound inositolpolyphosphate 5 phosphatase. AB - Membrane-bound inositolpolyphosphate 5-phosphatase was solubilized and highly purified from a microsomal fraction of rat liver. Its physiochemical and enzymological properties were compared with those of highly purified preparations of two types of soluble enzyme (soluble Type I and Type II) from rat brain. The molecular masses of the membrane-bound and soluble Type I enzymes were 32 kDa, while that of soluble Type II enzyme was 69 kDa, as determined by molecular sieve chromatography. The membrane-bound and soluble Type I enzymes showed similar broad peaks on isoelectric focusing (pI 5.8-6.4), while soluble Type II enzyme showed multiple peaks in the region between pI 4.0-5.8. All three enzymes required divalent cation for activity. Mg2+ was the most effective for both the membrane-bound and soluble Type I enzymes, while Co2+ enhanced soluble Type II enzyme activity about 1.5-fold relative to Mg2+ at 1 mM. The optimal pH of both the membrane-bound and soluble Type I enzymes was 7.8, while that of soluble Type II was 6.8. The Km values for inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] of all three enzymes were similar (5-8 microM), but those for inositol 1,3,4,5 tetrakisphosphate [Ins(1,3,4,5)P4] were quite different, the Km values of membrane-bound and soluble Type I enzymes being 0.8 microM, while that of soluble Type II was 130 microM. These similarities between the membrane-bound and soluble Type I enzymes suggest that these two molecules may be the same protein, and that concentrations of Ins(1,4,5)P3 and Ins(1,3,4,5)P4, both of which are considered to play critical roles in the regulation of intracellular Ca2+-concentration, may be differently regulated by two functionally distinct enzymes. PMID- 2558113 TI - Immunochemical and biochemical distinction of subtypes of atrial natriuretic peptide receptor. AB - The presence of subtypes of atrial natriuretic peptide (ANP) receptor has been demonstrated by examining the immunological features and ligand specificities of the receptors in various bovine tissues. The antibody probe used was the antiserum raised against the bovine lung ANP receptor, and the tissues examined for the possible presence of different types of ANP receptor were the lung, kidney, adrenal cortex, ovary, choroid plexus, and vascular tissues. When incubated with Triton extracts of these tissues, the antiserum strongly cross reacted with the ovary, kidney, and choroid plexus receptors as well as the homologous lung receptor (type I). The adrenal and vascular receptors were recognized only weakly, however, suggesting the presence of distinct ANP receptors (type II). In support of this immunochemical subtyping, type I and type II receptors showed a marked difference in their ability to bind the ANP analog atriopeptin I (ANP5-25): type I receptors in the lung exhibited a moderate affinity for atriopeptin I with a KD of 10(-9) M; however, type II receptors in adrenal and artery showed only a weak affinity for the analog with a KD of 10(-6) M. Structural analysis of affinity-labeled ANP receptors by SDS-PAGE indicated that type I and type II receptors have similar disulfide-linked dimeric structures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558115 TI - Phosphorylation of the 27-kDa gap junction protein by protein kinase C in vitro and in rat hepatocytes. AB - We previously demonstrated that the 27-kDa major component protein in rat liver gap junctions was phosphorylated by protein kinase C in vitro (Takeda, A. et al. (1987) FEBS Lett. 210, 169-172). In this study, we examined this further and examined the phosphorylation of the 27-kDa gap junction protein in rat hepatocytes by metabolically labeling cells with [32P]orthophosphate and using a monoclonal antibody to immunoprecipitate the protein. The in vitro phosphorylation was inhibited by monoclonal antibodies recognizing the carboxyl- (C-)terminal domain of the 27-kDa protein. Protease digestion analysis revealed that phosphorylation occurred at the C-terminal domain. In rat hepatocytes, the phorbol esters, 12-O-tetradecanoylphorbol-13-acetate and phorbol-12,13 dibutyrate, stimulated the 27-kDa protein phosphorylation, whereas 4 alpha phorbol-12,13-didecanoate did not. 1-Oleoyl-2-acetyl-sn-glycerol also stimulated the 27-kDa protein phosphorylation. In addition, norepinephrine stimulated the phosphorylation and pretreatment of hepatocytes with staurosporine, a potent inhibitor of protein kinase C, inhibited this stimulatory effect of norepinephrine. Both in vitro and in hepatocytes, analysis of chemical cleavage of the 27-kDa phosphoprotein revealed that phosphorylation occurred mainly at a 10-kDa fragment which the antibodies recognized. These results indicate that protein kinase C phosphorylates the 27-kDa gap junction protein, not only in vitro but also in hepatocytes, at the C-terminal domain of the protein. PMID- 2558116 TI - Toxic effects of aldehydes released from fixed pericardium on bovine aortic endothelial cells. AB - Glutaraldehyde (GA) and formaldehyde (FA) were shown to be released from 1 cm2 fixed pericard patches into 2-mL storage solutions after three 2-min washings in concentrations of about 1 mg/L. The cytotoxicity of these aldehyde concentrations on bovine aortic endothelial cells (BAECs) was evaluated in vitro, by proliferation capacity, cellular ATP content, PGI2 release and cyclic AMP synthesis. Continuous incubation of BAECs with GA greater than 0.1 mg/L and FA greater than 0.5 mg/L resulted in a significantly inhibited proliferation and in an increase of the intracellular Ca2+ triggered parameters, PGI2 and cyclic AMP, up to three fold. This strongly suggests an aldehyde-induced inhibition of the plasma-membrane bound Ca2+-ATPase, an enzyme which normally maintains low intracellular Ca2+-level. From these findings there is evidence that aldehydes released from bioprosthetic valve tissue may contribute to the lack of endothelial cell coverage in human implants. PMID- 2558117 TI - Tetanus toxin as a marker for small-cell lung cancer cell lines. AB - Tetanus toxin labeling of human lung cancer cell lines was investigated using direct and indirect immunofluorescence and immunohistochemical staining. Cells of characterized permanent cell lines, eight small-cell lung cancer (SCLC) cell lines of classic subtype, six SCLC cell lines of variant subtype and seven non small-cell lung cancer (NSCLC) cell lines, were incubated with a saturating concentration of tetanus toxin. For staining, fluorescein-isothiocyanate-(FITC) conjugated anti-(tetanus toxin) antibodies were used or a mouse monoclonal anti (fragment C) antibody with subsequent binding of FITC-conjugated anti-(mouse Ig) antibody or peroxidase-anti-peroxidase complex. Only SCLC showed an intense fluorescence/immunoreactivity restricted to the cell membrane. Quantitative analysis of tetanus toxin labeling by flow cytometry revealed the percentage of positive cells to be between 35% and 95% in SCLC without obvious differences between the classic and variant subtypes of SCLC. In NSCLC the percentage of positive cells was lower than 10%. These results demonstrate that SCLC in contrast to NSCLC can be labeled with tetanus toxin, emphasizing the neuroendocrine properties of this tumor, and that tetanus toxin labeling may become a useful diagnostic marker for SCLC cells in cytology. PMID- 2558118 TI - Plasma glutamate levels, lymphocyte reactivity and death rate in patients with bronchial carcinoma. AB - Elevated glutamate concentrations are commonly observed in patients with advanced carcinoma, and glutamate was recently found to inhibit the membrane transport of cystine and to impair the function of macrophages and lymphocytes in vitro. We therefore investigated the possibility that elevated plasma glutamate levels may be quantitatively correlated with reduced lymphocyte reactivity and an impaired host response to the tumor. Here we report the results of a study on patients with bronchial carcinoma, which show that patients with plasma glutamate levels above 120 microM have a lower lymphocyte response to mitogens and a substantially higher death rate than those with glutamate levels below 120 microM. This correlation does not prove a causal role of glutamate, but it confirms predictions from the in vitro laboratory data. PMID- 2558119 TI - Protection of melanoma cells against superoxide radicals by melanins. AB - Human melanoma cells transplanted into immunocompetent mice by the 6-day subrenal capsule technique are characterized by high resistance against immunological attack. This resistance is suggested to be the consequence of scavenging of superoxide free radicals by melanin. Scavenging of superoxide radicals by the melanoma cells was clearly demonstrated using electron spin resonance techniques. From comparison with synthetic melanins it is concluded that the scavenger effect can be attributed mainly to low-molecular-mass melanins synthesized in the melanoma cells whereas high-molecular-mass melanins are practically ineffective. PMID- 2558120 TI - Changes in nuclear distribution of large tumor antigen correlate with the proliferative activity of simian virus 40-transformed cells. AB - A correlation between intranuclear distribution of large tumor antigen (T-ag) and proliferative activity in Simian virus 40 (SV40)-transformed cells has been established. Nuclear T-ag was cytochemically detected by the immunoperoxidase reaction. Cell populations with a high growth rate displayed a heterogeneous pattern with patches of positive reaction surrounded by large and irregular negative areas. By contrast, cell populations with a low growth rate presented a rather homogeneous distribution of nuclear T-ag with a finely granular positive reaction all over the nucleus, excluding nucleolus-like areas. The latter pattern was also observed in cells that had been inhibited from traversing S phase. In this case, the heterogeneous distribution of T-ag within the nucleus was recovered once the cells were allowed to proceed again through S phase, or through G2 phase. Highly condensed chromatin in mitotic chromosomes appeared to exclude T-ag fully as early as in prophase. Then, nuclear distribution of T-ag changes throughout the cell cycle, thus suggesting it might be a cell cycle dependent event. Interestingly, thymidine-labeled cells from cultures with a high growth rate presented a heterogeneous distribution of autoradiographic grains. Regardless of their mechanism(s) of origin, T-ag-enriched areas might correspond to nuclear sites where a specific function, such as DNA synthesis, is being carried out. PMID- 2558123 TI - [Leukotriene B4, leukotriene C4 and prostaglandin E2 in the serum, synovial fluid and synovium in patients with rheumatoid arthritis]. AB - This study investigated the presence of arachidonic acid metabolites, (leukotriene B4, leukotriene C4 and prostaglandin E2) in the serum, synovial fluid and synovium of rheumatoid arthritis patients (RA). We obtained the serum, synovial fluid and synovium from 16 female patients with RA during knee operations. Leukotriene B4 (LTB4), leukotriene C4 (LTC4) and prostaglandin E2 (PGE2) were detected by RIA analysis. In the patients administered steroids, a positive correlation was found between the concentrations of LTB4 and PGE2 in the synovium. In the patients not administered steroids, significant correlations were found between the concentrations of LTB4 in the synovium and serum, and also between the concentrations of LTB4 and LTC4 in the synovium. The levels of immunoglobulins and LTB4 in the serum were determined and a significant correlation was found between the content of IgG and the concentration of LTB4. In the serum, the concentration of LTB4 was elevated in RA patients relative to the control normal subjects, and the difference was statistically significant. In conclusion, these data suggest that LTB4 in the serum reflects the concentration of LTB4 in the synovium. PMID- 2558122 TI - Inhibitors of topoisomerases I and II arrest DNA replication, but do not prevent nucleosome assembly in vivo. AB - Specific inhibitors of eukaryotic DNA topoisomerases I and II (camptothecin and VM-26, respectively) were used to examine the involvement of topoisomerases in DNA replication and chromatin assembly in vivo. When used singly, either camptothecin or VM-26 inhibited DNA synthesis in HeLa cells by more than 80%; when used simultaneously, the inhibitors effectively stopped replication, demonstrating that at least one class of topoisomerase must be active for fork propagation in vivo. To study nucleosome assembly during topoisomerase inhibition, three experimental strategies were employed: (1) pulse-chase experiments; (2) analyses of chromatin synthesized during residual replication in the presence of either camptothecin or VM-26; and (3) the assembly of previously replicated, unassembled DNA, generated in the presence of protein synthesis inhibitors. Using sensitivity to micrococcal nuclease and the maturation of non nucleosomal replication intermediates as criteria, neither camptothecin nor VM 26, alone or in concert, inhibited nucleosome assembly under any experimental protocol tested. These data provide evidence that, although topoisomerase activity is essential for DNA replication, neither continuous fork propagation nor topoisomerase activity is required for chromatin assembly on new DNA. PMID- 2558121 TI - Inositol tris- and polyphosphate formation during chemotaxis of Dictyostelium. AB - Using the technique of HPLC with Partisil SAX columns, we have found that stimulation of amoebae of Dictyostelium discoideum with the chemoattractant cyclic AMP induces the rapid accumulation of inositol 1,4,5-trisphosphate (Ins(1,4,5)P3), with a peak at 5 s. A smaller HPLC peak (designated P3) that elutes just after the Ins(1,4,5)P3 peak accumulates more slowly to a maximum at 20 s. In control studies, the changes in Ins(1,4,5)P3 were shown not to be due to varying recovery from the cell extracts and a comparison of reverse-phase and Partisil SAX HPLC columns showed similar values for determinations by either method. The involvement of a G-protein in this chemotactic system was confirmed by the finding that accumulation of Ins(1,4,5)P3 was elicited by the addition of GTP gamma S (5'-[gamma-thio]triphosphate) to saponin-permeabilized amoebae. A study of the changes in the lipid-soluble phosphatidyl inositol phosphates demonstrated that cyclic AMP also stimulated a rapid loss of radioactivity from 32P-labelled phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2), which corresponded in its timing to the rise in Ins(1,4,5)P3, indicating that a phosphoinositidase C (phospholipase C) is present that can be stimulated by occupation of the cell surface cyclic AMP receptors. PMID- 2558124 TI - Immunohistochemical localization of Na+, K+-ATPase in human normal and malignant pancreatic tissues. AB - Localization of Na+, K+-ATPase in the human pancreas was investigated immunohistochemically using rabbit antisera against Na+, K+-ATPase of the human kidney. The reaction product existed only on the luminal surfaces of both centroacinar and ductal cells in normal pancreatic tissue, whereas in chronic pancreatitis the localization of Na+, K+-ATPase was found frequently on the luminal surfaces of both centroacinar and ductal cells, and on the basolateral surfaces of some ductal cells. However, in acinar cells, the distribution of Na+,K+-ATPase was not detected in either the normal pancreas or chronic pancreatitis. In pancreatic carcinoma tissues, Na+,K+-ATPase existed very rarely in malignant cells. These results indicate that Na+,K+-ATPase is immunohistochemically localized on the membranes of centroacinar and ductal cells of the human pancreas, and that the antigenicity of Na+,K+-ATPase in pancreatic carcinoma cells differs from that in normal cells. PMID- 2558125 TI - [Angiographic study of peripheral lung field mass lesions by superselective pulmonary arteriography]. AB - Studies of pulmonary arteriography have a long history in the diagnosis of lung tumors (Steinberg and Robb, 1938). The main purpose of these studies was mostly the determination of the resectability of neoplasms. So that the role of pulmonary arteriography on the diagnosis of peripheral lung tumors has yet been obscure. Neither conventional nor selective pulmonary arteriography can clearly visualize vascular structures of mass lesions in the peripheral lung fields, because of overlaid other segmental arteries, dilution of the contrast medium and blood flow shift. In the present study, direct catheterizations to the segmental branches of peripheral lung field mass lesions were performed on 62 cases, in order to visualize and investigate more detailed vascular structures of them. Twenty nine cases were of primary lung cancer; 14 of metastatic and 19 of inflammatory mass. By means of the superselective pulmonary arteriography, vascular wall irregularities and tumor stain were frequently observed in the cases of malignancies. Either occlusion of minute branches (1 mm greater than or equal to) or encasement of pulmonary arterial branches (1 mm less than) was seen more frequently in the cases of primary lung cancers (86%) than in the cases of metastatic lung cancers (64%). Tumor stain was also observed both in the cases of primary lung cancers (55%) and metastatic lung cancers (64%). In the cases of primary lung cancers, tendency for visualization of vascular wall irregularities and tumor stain seemed to depend upon rather their histological types and differentiation grades than the size of the mass. Encasement or obstruction of the arterial branches (1 mm less than) was seen more frequently in the cases of poorly differentiated adenocarcinoma, moderately differentiated squamous cell carcinoma and anaplastic carcinoma than in the cases of other histologic types. On the other hand, tumor stain was observed more frequently in the cases of moderately differentiated adenocarcinoma and anaplastic carcinoma than in the cases of other histologic types. Obstruction of arterial branches (1 mm less than) was seen also in a few cases of the inflammatory masses, but that of inflammatory masses was usually distinguished from that of malignant masses by their characteristic surrounding vascular natures such as central vascular convergence, widespread simple narrowing of arterial branches and diminution or lack of ramification of minute branches. Tumor stain was not observed in the cases of inflammatory masses. PMID- 2558126 TI - Improvement of survival and fertilizing capacity of human spermatozoa in an IVF programme by selection on discontinuous Percoll gradients. AB - Two techniques for the separation of spermatozoa were compared: swim-up migration (SUM) and centrifugation on a discontinuous Percoll gradient (CPG). Their respective effects on sperm motility were analysed by computer-assisted videomicrography in either normal or asthenozoospermic groups. In both groups, there was no difference in any of the motion parameters between the two treatments after 1-h incubation. However, a clear difference was observed after 24 h when excellent motility was retained only in the CPG-treated group. A total of 350 ejaculates were produced by the husbands of women undergoing oocyte retrieval in an IVF programme. Spermatozoa were treated by CPG when the infertility was due to poor quality spermatozoa (n = 91), when there was a known previous history of semen infection (n = 73) or when frozen semen, originating from a donor, was used (n = 36). In all other cases (n = 150), spermatozoa were treated by SUM. The cleavage rates obtained were 32.2, 70.1, 60.9 and 68.6% respectively in the four categories. The clinical pregnancy rates per oocyte retrieval were 19.8, 31.5, 22.2 and 18.0% respectively. Forty-eight births occurred in the CPG group: 28 boys and 20 girls, all normal. We conclude that CPG is useful, both in cases of poor semen quality and in tubal infertility, in which the clinical pregnancy rate increased significantly from 18.0 to 31.5%. PMID- 2558127 TI - Two rapid and simple enzyme immunoassays for human antibodies to Entamoeba histolytica. AB - Two rapid and simple enzyme immunoassays (EIA) for antibodies to E. histolytica the protozoa causing ambiasis, are described. In the rapid dot EIA, a qualitative procedure, antigens were dried as a small dot (3 mm in diameter) on a thin white opaque polystyrene strip and serum samples were assayed undiluted. The assay required 3 incubation periods, 1 to 3 minutes each, and was completed in 9 minutes, with a positive reaction revealed as a blue color (precipitate) on the antigen dot and negative as colorless. The developed color is stable for permanent record. In the Microwell EIA, a quantitative procedure, antigens were dried in the Microwells. The assay also consisted 3 incubation periods of 15 minutes each, and was completed in 50 minutes. The results in absorbance values were normalized to EIA units (EU). Both tests had good reproducibility, sensitivity and specificity; and highly correlated with 3 other serologic tests. Their reagents can be stored for more than a year. Both tests could be suitable for small and physicians' office laboratories, especially in developing countries. PMID- 2558128 TI - Comparative pathology of canine mammary tumours. AB - The pathological features of 52 canine mammary tumours were studied and compared with those of human breast neoplasms. In many of the former, the constituent cells were both epithelial and myoepithelial. Cartilaginous and osseous metaplasia of the stromal tissue was striking in a few tumours. Carcinosarcomas were also encountered. On a careful search, virus-like particles were observed in five tumours. The similarities and differences between the human and canine mammary tumours were discussed with a view to assessing the suitability of dog mammary tumour as an animal model for human breast tumours. PMID- 2558130 TI - Hemophagocytic syndrome associated with coxsackie virus A 9 infection in a non immunosuppressed adult. PMID- 2558129 TI - An aortic body carcinoma with multifocal thoracic metastases in a cat. AB - A necropsy performed on an aged cat with respiratory signs revealed a large white firm mass cranioventrally to the heart, and multiple similar masses scattered throughout the lungs, diaphragm, pericardial sac and intercostal muscles. The histological examination of the primary site and the metastases are consistent with a well-differentiated aortic body carcinoma. PMID- 2558131 TI - Rapidly enlarging gingival mass. AB - A case of small cell carcinoma metastatic to the mandible has been detailed. The lesion involved both hard and soft tissues and it was impossible to determine the site to which it first spread. Because of the histological similarity to the primary lung lesion and evidence of metastatic spread to other body organs, this lesion was considered to be metastatic rather than a primary tumor. The importance of accurate diagnosis was discussed, and treatment options were presented. PMID- 2558133 TI - Immunodetection of cathepsin D in neuritic plaques found in brains of patients with dementia of Alzheimer type. AB - The aspartic endopeptidase cathepsin D was immunolocalized in 21 human cadaver brains from patients with dementia and controls. Immunoreactive cathepsin D was found to be present in multiple neurons, neuritic plaques, some macroglial cells, and microvessels. It is suggested that the enzyme might be involved in certain posttranslational changes of cystoskeletal compounds that lead to the formation and/or further growth of neuritic plaques and neurofibrillary tangles. PMID- 2558132 TI - The effect of acute hypoxaemia on ventricular function during beta-adrenergic and cholinergic blockade in the fetal sheep. AB - The effect of acute hypoxaemia on right and left ventricular function was investigated in 8 fetal sheep (137-140 days gestation). Fetuses were instrumented with electromagnetic flow sensors on the ascending aorta and the main pulmonary artery. After 8 days recovery, hypoxaemia was achieved by reducing the maternal ewe's inspired O2 concentration to 13.1 +/- 1.5%. Control and hypoxaemic arterial blood values were pH 7.37 +/- 0.04 (SD) and 7.35 +/- 0.06, PCO2 48.0 +/- 2.8 and 47.6 +/- 5.1 mmHg, PO2 19.9 +/- 2.2 and 11.4 +/- 1.5 mmHg, haematocrit 37.5 +/- 1.2 and 39.5 +/- 2.2, respectively. Arterial pressure increased insignificantly with acute hypoxaemia (50.2 +/- 3.9 to 53.6 +/- 8.1 mmHg). Left and right ventricular performance was assessed by generating biventricular function curves relating stroke volume to mean atrial pressure. All function curves were composed of steep ascending and plateau limbs that intersected at a breakpoint. Comparing control and hypoxaemia function curves, the left ventricular stroke volume breakpoints were 0.79 +/- 0.20 and 0.78 +/- 0.21 ml/kg, respectively, while the right ventricular stroke volume breakpoints were 0.99 +/- 0.11 and 0.88 +/- 0.21 ml/kg (n.s.). In 4 fetuses, acute hypoxaemia was associated with significant increases in arterial blood pressure (P less than 0.05). In these fetuses, the right ventricular function curve was shifted significantly downward compared to the control right ventricular curve. When nitroprusside was given to these hypertensive fetuses to return blood pressure to control levels, the right ventricular function curve returned to baseline. We conclude that even under conditions of extreme hypoxaemia, ventricular function is well preserved in the normotensive fetal sheep. However, when increases in arterial pressure also accompany hypoxaemia, detectable changes in right ventricular function can be accounted for by changes in arterial pressure. PMID- 2558134 TI - Beta-adrenergic receptor differences in cultured arterial smooth muscle cells between spontaneously hypertensive and Wistar-Kyoto rats. AB - In this study, beta-adrenergic receptors of cultured arterial smooth muscle cells (ASMC) from normotensive Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) were compared. Scatchard analysis of [3H]-dihydroalprenolol hydrochloride (DHA) binding revealed that the maximum binding capacity (Bmax) for [3H]-DHA binding was significantly higher in WKY than in SHR, by twofold (n = 6, P less than 0.0001). Analysis of isoproterenol competition for [3H]-DHA binding by computerized non-linear regression indicated that the two strains have an equal number of high-affinity-state receptors, but that WKY rats have about three times more low-affinity-state receptors than SHR. Isoproterenol stimulated cyclic adenosine monophosphate (cAMP) production in a dose-dependent manner, and dose response experiments revealed an effective concentration of ligand at which 50% of the maximum effect is observed (EC50) of 5-10 x 10(-8) mol/l isoproterenol for both WKY and SHR. Also, isoproterenol-stimulated cAMP production was inhibited by propranolol in a dose-dependent manner. This study demonstrates that the previously described difference observed for in vivo binding of beta-adrenergic receptors of vessels from SHR is retained in tissue culture of vascular smooth muscle and is, therefore, independent of the elevated plasma catecholamine and blood pressure levels found in SHR. The finding that there is equal cAMP stimulation in WKY and SHR is consistent with the discovery of an equal number of high-affinity-state receptors in the two strains. The difference in total beta adrenergic receptor number is secondary to the greater number of low-affinity state receptors found in WKY. PMID- 2558135 TI - Antihypertensive contribution of sodium depletion and the sympathetic axis during chronic angiotensin II converting enzyme inhibition. AB - The known physiological adaptation of cardiovascular sensitivity to variations in angiotensin II (Ang II) levels would predict that the blood pressure (BP) lowering effect of Ang II inhibition might be at least partly counterbalanced by enhanced Ang II reactivity. Therefore, factors other than Ang II inhibition per se may contribute to the antihypertensive mechanisms of angiotensin converting enzyme (ACE) inhibitors. In order to further investigate this, the body sodium blood volume state as well as the pressor reactivity to infused Ang II or norepinephrine (NE) were assessed in 12 normal subjects and 16 patients with essential hypertension given a placebo, and after 6 weeks of intervention with enalapril (20-40 mg/day). Enalapril produced in both groups similar falls in plasma ACE activity (P less than 0.0001) and upright plasma aldosterone (P less than 0.01), and a rise in plasma renin activity (PRA; P less than 0.05). BP decreased from 156/107 +/- 3/2 (mean +/- s.e.m.) to 142/94 +/- 5/3 mmHg (P less than 0.001) in the hypertensives and from 118/84 +/- 4/2 to 111/73 +/- 4/3 mmHg (P less than 0.01) in the normal subjects. In the hypertensive patients only, the Ang II pressor reactivity relative to Ang II plasma levels during Ang II infusion was increased (P less than 0.01), while the NE pressor reactivity relative to NE plasma levels during NE infusion (P less than 0.01) as well as the exchangeable body sodium (-5%, P less than 0.001) were reduced significantly. Blood and plasma volume, levels of plasma atrial natriuretic factor and catecholamines, and the heart rate and its response to isoproterenol were unchanged in both groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558136 TI - A hybrido-immunocytochemical assay for the in situ detection of cytomegalovirus DNA using digoxigenin-labeled probes. AB - A non-radioactive hybrido-immunocytochemical assay for the detection of cytomegalovirus (CMV) DNA in infected cells was developed. Two different DNA fragments belonging to the repeated sequences of CMV genome were used to construct the hybridization probe. The probe was constructed by incorporating deoxyuridine triphosphate labeled with digoxigenin. The in situ hybridized CMV DNA probe was immunocytochemically visualized by anti-digoxigenin. Fab fragments labeled with alkaline phosphatase. This procedure permitted the DNA detection, in the nuclei of infected cells fixed at 48 h after infection, of the Towne CMV reference strain and 21 different laboratory-isolated CMV strains. Our assay demonstrated a high specificity, sensitivity and reproducibility. PMID- 2558137 TI - A microplate immunofluorescence assay for the detection of monoclonal antibodies against viral antigens in cell culture. AB - A microplate immunofluorescence assay for the screening of culture fluids from large numbers of hybridoma clones has been developed. This method enables the simultaneous screening of a small volume of culture fluid from initial hybridoma clones on several different viruses or several strains of the same virus, enabling the early isolation and stabilization of the clones with the desired specificity. PMID- 2558138 TI - Catalyzed reporter deposition, a novel method of signal amplification. Application to immunoassays. AB - A novel signal amplification method, catalyzed reporter deposition (CARD), and its application to immunoassays is described. The method involves utilizing an analyte-dependent reporter enzyme (ADRE) to catalyze the deposition of additional reporter on the surface in a solid-phase immunoassay. In the examples described, deposition of reporter is facilitated by using a horseradish peroxidase (HRP) ADRE to catalyze the deposition of biotin labeled phenols. The deposited biotins are then reacted with streptavidin-labeled enzyme, thereby resulting in deposition of enzyme. Using the ADRE to catalyze the deposition of additional enzyme results in an amplification of the signal of the ADRE alone and improves the detection limit of the assay. The method is highly sensitive, simple, flexible, and easy to implement. PMID- 2558139 TI - The influence of adsorption of native and modified antibodies on their activity. AB - Immobilization of biomolecules to solid phase materials has been widely used in many areas (e.g., purification, analytical chemistry, and catalysis). The interfacial properties of immobilized antibodies on pretreated silica and hydrogel surfaces were explored by comparing native and modified antibodies with respect to their surface activity. The antibody was modified by exposing it to a low pH solution prior to immobilization. Both physical adsorption and covalent immobilization methods were studied. It was found that the surface activity of the modified antibody is higher than that of the native antibody on two silica surfaces. The results of this study demonstrate that the adsorption properties of the antibodies play an important role in their covalent immobilization on certain types of solid supports. PMID- 2558141 TI - Uncoupling of receptor-mediated cellular responses by ionic lithium. AB - The propensity of ionic lithium to interfere with the coupling of receptors to guanine nucleotide binding proteins (G-proteins) has only recently been investigated using rat cortical membranes. In the present study we have used intact isolated osteoclasts to investigate lithium-induced uncoupling of the receptor-mediated actions of calcitonin. All actions of calcitonin on the osteoclast were abolished by ionic lithium. We believe that the cation prevents signal transduction by inhibiting G protein-receptor interaction, the first step in intracellular signalling. PMID- 2558140 TI - Malignant fibrous histiocytoma of the nasopharynx. AB - Malignant Fibrous Histiocytomas (MFH) are uncommon in the head and neck and extremely rare in the nasopharynx. We report a case of MFH arising in the nasopharynx and describe its diagnosis and management. The literature is reviewed and the likely origins of the tumour and its behaviour are discussed. PMID- 2558142 TI - The hunt for the CIA: factors which demonstrate corticotrophin-inhibitory activity. PMID- 2558143 TI - Leydig cell recovery following a second challenge with ethane-1,2 dimethanesulphonate is enhanced by short intervals between cytotoxin administration to rats. AB - Ethane-1,2-dimethanesulphonate (EDS) destroys Leydig cells in the testis of the adult rat and subsequently a new population of Leydig cells develops. It has been reported that EDS is not cytocidal to the new immature Leydig cell population. In the present study, the effect of increasing the time-interval between injections of EDS on cytotoxicity to Leydig cells was examined. At time-intervals of 4-10 weeks between injections the response was similar to that seen after a single injection of EDS to the adult rat. Four days after the second injection, EDS was found to reduce substantially serum testosterone concentrations and in-vitro binding of 125I-labelled human chorionic gonadotrophin (hCG) to testicular LH receptors which can be correlated with Leydig cell destruction. However, when the interval was only 2 or 3 weeks there was no reduction in serum testosterone, and 125I-labelled hCG binding was not so markedly reduced. During days 1-6 after a second injection of EDS, administered 3 weeks after the first, there were marked reductions in serum testosterone concentrations and in 125I-labelled hCG binding to testis homogenates within 24 h. Recovery from the effects of EDS was rapid, and increased Leydig cell activity was seen from 2 to 6 days after injection. In contrast to the established changes in the adult rat, there was only a 50% reduction in the number of Leydig cells positive for 3 beta-hydroxysteroid dehydrogenase 2 days after the second injection of EDS, and after 6 days the number of cells had increased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558144 TI - Specific effect of oestrone sulphate on protein synthesis and secretion by cultured epithelial cells from guinea-pig endometrium. AB - Patterns of induced protein synthesis and secretion in guinea-pig endometrial epithelial cell cultures in response to oestrone sulphate alone and oestrone sulphate plus progesterone were investigated. Epithelial cells were cultured for 3 days in growth medium, then washed three times in a steroid-free medium. For each experiment, anticytokeratin immunostaining was used to discriminate the epithelial cells from the stromal cells. Only experiments in which the control dishes displayed more than 80% of anticytokeratin-immunostained cells were further processed. After this period oestradiol-17 beta (20 nmol/l; control), oestradiol-17 beta (20 nmol/l) plus progesterone (0.5 mumol/l), oestrone sulphate (1 mumol/l) or oestrone sulphate (1 mumol/l) plus progesterone (0.5 mumol/l) were added to the medium for 48 h. An immunocytochemical progesterone receptor assay showed that oestradiol-17 beta increased the progesterone receptor content of cells, and progesterone added to cultured cells in the presence of oestradiol-17 beta induced a significant increase in oestrogen sulphotransferase activity assessing the hormone responsiveness of the cultured cells. In these culture conditions and after 16 h of incubation, oestradiol-17 beta induced a 1.7-fold increase in [3H]thymidine incorporation into DNA, and [35S]methionine incorporation into cellular proteins was linearly increased up to 8 h. Biochemical changes induced by the different hormone treatments were studied by labelling the proteins with a 6-h pulse of [35S]methionine. The proteins present in the medium and in cells were analysed by two-dimensional polyacrylamide gel electrophoresis, followed by fluorography.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558145 TI - Regulation by oestradiol of serum levels of LH and FSH and pituitary levels of gonadotrophin-releasing hormone receptors after ovariectomy in the rat. AB - We studied cyclic and ovariectomized, oestradiol treated rats to investigate whether oestradiol concentrations before ovariectomy determine the dynamics of the rises in serum levels of LH and FSH, and in pituitary gonadotrophin-releasing hormone (GnRH) receptors after ovariectomy. In cyclic rats, ovariectomy on metoestrus (oestradiol = 44 pmol/l), but not at midnight of dioestrus (oestradiol = 254 pmol/l) was followed by a rise in GnRH receptors 40 h later. Randomly cyclic females were ovariectomized under ether anaesthesia and treated with s.c. oestradiol implants for 2 days to provide serum levels spanning the physiological range. Exposure to lower oestradiol concentrations (48 or 74 pmol/l) resulted in significant rises in levels of LH and FSH, and in GnRH receptors earlier than exposure to higher (148 or 309 pmol/l) oestradiol levels (2 compared with 4 days). GnRH given for 24 h after removal of the implant was unable to induce a rise in GnRH receptors in any group, while LH and FSH responses to GnRH were proportional to the pretreatment levels of oestradiol. This suggests that the concentration of oestradiol before ovariectomy determines the subsequent dynamics of LH and FSH and of GnRH receptors through actions at the hypothalamic and pituitary level. PMID- 2558146 TI - Ontogeny of ACTH(1-24) receptors in rat adrenal glands during the perinatal period. AB - Binding of ACTH to receptors was studied on crude adrenal membranes from fetal and newborn rats. 125I-Labelled ACTH(1-24) was used as the radioligand, the steroidogenic potency of which was 100-fold lower than that of unlabelled ACTH(1 24). Binding was specific, rapidly equilibrated and temperature dependent. Scatchard analysis of the binding data revealed a single class of binding sites with a dissociation constant of about 100 nmol/l at all stages of development studied. The concentration of ACTH receptors expressed per mg membrane proteins decreased in fetuses between days 17 and 21 of gestation and remained stable in newborn rats from weeks 1 to 4. The number of ACTH receptors expressed per adrenal increased regularly in fetal and newborn rats. The perinatal evolution of these concentrations of ACTH receptors is related to the increase in the size of the adrenals and the changes in cytoplasmic structures of the adrenocortical cells. When the number of ACTH-binding sites was expressed per microgram DNA, maximum values occurred in fetuses on day 19 of gestation, and minimum values in newborn rats, 1 week after birth. There was an excellent correlation between the plasma levels of immunoreactive ACTH and corticosterone and the number of ACTH receptors per microgram DNA during the perinatal period. Other results suggest that ACTH is able to up-regulate the number of its own receptors. PMID- 2558147 TI - Differential activation of the pituitary-adrenocortical axis after stress in the rat: use of two genetically selected lines (Roman low- and high-avoidance rats) as a model. AB - We have examined the activation of the pituitary-adrenal axis in two lines of rats, the Roman high (RHA)- and low (RLA)-avoidance rats known to be emotionally different. These rats are selected for rapid acquisition of a conditioned avoidance response (RHA) compared with failure to acquire this response (RLA). In this study the endocrine response (ACTH, corticosterone, aldosterone) of RLA and RHA rats to two types of stress was examined: exposure to open-field stress for 10 min (Op) or exposure to ether vapours for 3 min (E). Basal plasma ACTH concentrations were lower in RLA than in RHA rats (RLA: 110.8 +/- 24.5 ng/l; RHA: 252.7 +/- 60.8 ng/l, P less than 0.05) but the absolute values of ACTH reached after both types of stress were comparable between RLA and RHA rats. Plasma corticosterone and aldosterone under resting conditions were not different between RLA and RHA rats. Plasma corticosterone was higher in RLA following openfield stress (P less than 0.05) while no differences between RLA and RHA were observed after ether stress (RHA: basal = 66 +/- 14.nmol/l, Op = 384 +/- 55, E = 606 +/- 75; RLA: basal = 121 +/- 52, Op = 612 +/- 92, E = 698 +/- 89). Stress induced increases in plasma aldosterone were higher in the RLA line after both types of stress (RHA: basal = 175 +/- 36 pmol/l, Op = 546 +/- 53, E = 563 +/- 47; RLA: basal = 272 +/- 64, Op = 1246 +/- 91, E = 863 +/- 72). PMID- 2558148 TI - Activity of 17 beta-hydroxysteroid oxidoreductase in tissues of the human fetus. AB - The interconversion of oestrone and oestradiol, androstenedione and testosterone, and dehydroepiandrosterone and 5-androstene-3 beta,17 beta-diol in mammalian tissues is catalysed by 17 beta-hydroxysteroid oxidoreductase (17 beta-HSOR). To identify tissue sites of 17 beta-HSOR activity in the human fetus, microsomal fractions from 15 different fetal tissues obtained from first and second trimester pregnancies were used for evaluation of enzymatic activity by use of [17 alpha-3H] oestradiol as the substrate and NADP+ as the co-factor. With these reagents, the enzyme-catalysed reaction led to the production of both non radiolabelled oestrone and NADP3H in equimolar amounts; the radioactivity associated with NADP3H was used to quantify 17 beta-HSOR activity. Activity of 17 beta-HSOR was present in microsomes of all the tissues evaluated. The specific activity of the enzyme was highest in liver and placental microsomes. The interconversion of oestradiol and oestrone in microsomal fractions of nine different fetal tissues was studied by the use of substrates labelled with tritium at stable nuclear positions ([6,7-3H]oestradiol and [6,7-3H]oestrone). The products, [3H]oestrone and [3H]oestradiol, were quantified by the use of established techniques; other metabolites formed in these incubations were not identified. The reductive pathway of metabolism (oestrone to oestradiol) appeared to be favoured in microsomal fractions prepared from placenta, fetal zone of the adrenal gland and, possibly, lung. The oxidative pathway (oestradiol to oestrone) appeared to be favoured in microsomes prepared from liver, intestine, stomach, kidney, brain and heart. 17 beta-HSOR activity in fetal liver also was assessed by the use of fresh and frozen-thawed tissue, homogenate, subcellular fractions, and, also, in primary hepatocytes maintained in culture; the specific activity of the enzyme was highest in the microsomal fraction of liver tissue and 17 beta HSOR activity in liver microsomes was linear with time of incubation up to 1 h. In hepatocytes, the enzymatic activity was linear with time of incubation up to 2 h and with cell number up to 2.5 x 10(5) cells/ml; the apparent Michaelis constant of hepatocyte 17 beta-HSOR for oestradiol was 11 mumol/l. The specific activity of 17 beta-HSOR did not change after pretreatment of hepatocytes for 24 h with insulin, glucagon or dexamethasone. PMID- 2558149 TI - Atrial natriuretic peptide protects hepatocytes against damage induced by hypoxia and reactive oxygen. Possible role of intracellular free ionized calcium. AB - Elevated concentrations of atrial natriuretic peptide reportedly mitigate acute renal failure in vivo and in the isolated perfused kidney (M. Nakamoto, J.I. Shapiro, P.F. Shanley, L. Chan & R.W. Shrier (1987) J. Clin. Invest. 80, 698-705; S.G. Shaw, J. Weidmann, J. Hodler, A. Zimmermann & A. Paternostro (1987) J. Clin. Invest. 80, 1232-1237). Since atrial natriuretic peptide has been shown to be a potent vasodilator, this beneficial effect may be due entirely to improved haemodynamics. To determine whether atrial natriuretic peptide also has a protective effect at the cellular level, rat hepatocyte cell cultures were treated with atrial natriuretic peptide prior to or after induction of cell damage by hypoxia (0.5% O2 for 4 h) or reactive oxygen (hypochlorous acid). Bleb formation, degradation of radiolabeled trichloroacetic acid-precipitable peptides, release of lactate dehydrogenase and trypan blue exclusion were used as indicators of cell damage. Atrial natriuretic peptide treatment distinctly protected the cell cultures against damage in both cases. This beneficial effect of atrial natriuretic peptide was partly mimicked by sodium nitroprusside, which, like atrial natriuretic peptide, largely increased the cellular cGMP content. 6 Anilino-5,8-quinolinedione (Ly 83583), an inhibitor of particulate guanylate cyclase, blocked the protective effect of atrial natriuretic peptide. Therefore a cGMP-mediated mechanism seems to be involved in the cytoprotective action of atrial natriuretic peptide. Fluorometric measurements using the Ca2+-sensitive dye Quin-2 showed that the elevation of intracellular Ca2+ after cellular insult by hypochlorous acid is prevented by atrial natriuretic peptide. These results suggest that atrial natriuretic peptide may attenuate hypoxic and toxic cell damage by increasing cGMP and reducing intracellular Ca2+. PMID- 2558150 TI - Early events in peptide-stimulated ecdysteroid secretion by the prothoracic glands of Manduca sexta. AB - The prothoracic glands of the tobacco hornworm, Manduca sexta, have been an advantageous model for investigating the cellular mechanisms underlying hormone stimulated ecdysteroid secretion in insects. The cerebral neuropeptide prothoracicotropic hormone (PTTH) is currently thought to activate the prothoracic glands via a calcium-dependent increase in cAMP synthesis, activation of cAMP-dependent protein kinase, and protein phosphorylation (Gilbert et al.: Bioessays, 8:153-158, '88). The present paper discusses current research regarding early changes in cell function elicited by PTTH, with emphasis on the regulation of cAMP synthesis and degradation and the involvement of translational events in PTTH action. PMID- 2558151 TI - A novel calcium current in dysgenic skeletal muscle. AB - The whole-cell patch-clamp technique was used to study voltage-dependent calcium currents in primary cultures of myotubes and in freshly dissociated skeletal muscle from normal and dysgenic mice. In addition to the transient, dihydropyridine (DHP)-insensitive calcium current previously described, a maintained DHP-sensitive calcium current was found in dysgenic skeletal muscle. This current, here termed ICa-dys, is largest in acutely dissociated fetal or neonatal dysgenic muscle and also in dysgenic myotubes grown on a substrate of killed fibroblasts. In dysgenic myotubes grown on untreated plastic culture dishes, ICa-dys is usually so small that it cannot be detected. In addition, ICa dys is apparently absent from normal skeletal muscle. From a holding potential of -80 mV. ICa-dys becomes apparent for test pulses to approximately -20 mV and peaks at approximately +20 mV. The current activates rapidly (rise time approximately 5 ms at 20 degrees C) and with 10 mM Ca as charge carrier inactivates little or not at all during a 200-ms test pulse. Thus, ICa-dys activates much faster than the slowly activating calcium current of normal skeletal muscle and does not display Ca-dependent inactivation like the cardiac L type calcium current. Substituting Ba for Ca as the charge carrier doubles the size of ICa-dys without altering its kinetics. ICa-dys is approximately 75% blocked by 100 nM (+)-PN 200-110 and is increased about threefold by 500 nM racemic Bay K 8644. The very high sensitivity of ICa-dys to these DHP compounds distinguishes it from neuronal L-type calcium current and from the calcium currents of normal skeletal muscle. ICa-dys may represent a calcium channel that is normally not expressed in skeletal muscle, or a mutated form of the skeletal muscle slow calcium channel. PMID- 2558152 TI - Human cytomegalovirus: recent aspects from molecular biology. PMID- 2558153 TI - Gene UL11 of herpes simplex virus type 1 encodes a virion protein which is myristylated. AB - We have investigated whether herpes simplex virus (HSV) contains structural polypeptides which are modified by myristic acid. We demonstrate that herpes simplex virions contain a family of myristylated proteins, Mr approximately 13,000 to 16,000. These were mapped, using HSV-1/HSV-2 intertypic recombinants, to 0.130 to 0.204 map units on the virus genome. Using anti-peptide sera, raised against the carboxyterminus of the predicted UL11 gene product, we have established that the myristylated virion polypeptides are products of the viral gene UL11. PMID- 2558154 TI - The unique N-terminal domain of the large subunit of herpes simplex virus ribonucleotide reductase is preferentially sensitive to proteolysis. AB - Using antisera made against peptides corresponding to different regions of the large subunit of herpes simplex virus type 1 ribonucleotide reductase we have probed proteolytic fragments of this protein and found that at least a part of its unique N-terminal domain is not necessary for enzyme activity. This non essential region encompasses the domain previously predicted to be composed of beta sheets with a well buried core of hydrophobic residues. Truncated forms of the large subunit are generated in vivo and are located almost exclusively in the nucleus. PMID- 2558155 TI - Presence of antibody to human herpesvirus 6 in monkeys. AB - A serological survey of monkeys was conducted to determine the prevalence of antibody to human herpesvirus 6 (HHV-6). Two-hundred and fifteen sera from 10 species of monkeys were examined by an immunofluorescent antibody (IF) assay. The antibody was found in monkeys from eight of the 10 species examined, but was not detected in silvered lutongs or cotton-top tamarins. The prevalence of antibody was highest in squirrel monkeys. Sera with high antibody titres were examined further by Western blot analysis and the neutralizing antibody test and the antibody levels were compared with that from a patient with exanthem subitum. On Western blotting, monkey and human sera that were antibody-positive to HHV-6 antigen gave similar reactions with antigen components of almost the same Mr. Furthermore, sera that were antibody-positive by the IF test were also positive by the neutralizing antibody test and their titres in the two tests were comparable. These results suggest the existence of HHV-6 or an HHV-6-related virus in monkeys. PMID- 2558156 TI - Preinfection prophylaxis with herpes simplex virus glycoprotein immunogens: factors influencing efficacy. AB - Using a guinea-pig model of genital herpes simplex virus (HSV) infection we explored the protection afforded by preinfection immunization with HSV glycoproteins. Glycoprotein immunogens prepared by recombinant DNA technology were found to be as effective as immunogens purified from HSV-infected cell cultures. Immunized animals developed less severe primary disease and also experienced less frequent recurrent infections. Protection was influenced by both adjuvant and route of administration. These studies suggest that recombinant HSV glycoproteins may be effective immunogens for human clinical trials, but that the development of an effective vaccine will require identification of new potent adjuvants that are safe for human use. PMID- 2558157 TI - Polymerase chain reaction for human picornaviruses. AB - We have used enzymic amplification of specific nucleic acid sequences followed by hybridization, for the rapid detection and typing of human picornaviruses after cell culture isolation. The test is based on the synthesis of cDNA, the polymerase chain reaction and the use of oligonucleotide probes. The primers were selected from the 5' non-coding region of the genome representing highly conserved regions. Sequences specific to enteroviruses and rhinoviruses were used as probes. The assay was able to identify all the picornavirus reference strains analysed and it was also possible to discriminate between enteroviruses and rhinoviruses by the hybridization procedure. When 29 picornavirus clinical isolates were analysed, all except one were detected by gel electrophoresis and a specific hybridization signal was obtained with all except three strains using the oligonucleotide probes. PMID- 2558158 TI - The nucleotide sequence of coxsackievirus A9; implications for receptor binding and enterovirus classification. AB - The complete nucleotide sequence of the genome of coxsackievirus A9 (CAV-9) has been determined from cDNA cloned in Escherichia coli. Excluding the 3' poly(A) stretch, the RNA genome is 7452 nucleotides long and encodes a single polyprotein of 2201 amino acids. Comparison of the nucleotide and predicted amino acid sequences with those of the coxsackieviruses B1, B3 and B4 reveals a surprising degree of homology, with overall amino acid homologies of 86.9%, 86.2% and 87.0%, respectively. In contrast, there is much less homology to another coxsackie A virus, CAV-21, 60.4% overall amino acid homology. This demonstrates the high degree of diversity within the CAV group and indicates that the current classification does not directly correlate with molecular genetic properties. One major feature of CAV-9 is an insertion, relative to all other enteroviruses sequenced to date, which is located at the C terminus of VP1, and includes an arginine-glycine-aspartic acid tripeptide. Such sequences in a number of other proteins are known to have activity in promoting attachment to cell receptors and the implications for CAV-9 receptor binding are discussed. PMID- 2558159 TI - Antigenic conservation of H1N1 swine influenza viruses. AB - Influenza viruses of the H1N1 subtype have been continually circulating in pigs in the U.S.A. for at least 50 years. To examine the level of antigenic variation in these swine viruses, a panel of 60 monoclonal antibodies (MAbs) to the haemagglutinin (HA) of recent swine isolates was prepared. Evaluation of neutralization escape mutants selected with these MAbs defined four antigenic sites on the HA, two of which overlap. Swine viruses isolated over 24 years in an enzootic area in Wisconsin were examined by ELISA and haemagglutination inhibition (HI) with these MAbs and the results indicated that the antigenic sites defined by these MAbs were highly conserved in these viruses. In comparing recent H1N1 viruses from pigs, turkeys, ducks and humans, changes in the antigenic sites were detected on the basis of HI reactivity. However, results of ELISA with these viruses clearly showed that the antigenic sites were still present on almost all H1N1 viruses of swine origin; thus, altered reactivity of these viruses in HI tests with MAbs was not a reflection of changes in the antigenic sites defined by the MAbs. It seems likely that the variation detected in these viruses occurs by a mechanism other than immune selection. PMID- 2558160 TI - Mutation of host cell determinants which discriminate between lytic and persistent mouse hepatitis virus infection results in a fusion-resistant phenotype. AB - The expression of mouse hepatitis virus (MHV) E2-specific mRNA, the E2 polypeptide and its associated cell fusing activity was monitored in various cell types inoculated with a recombinant vaccinia virus, designated vMS containing the E2 gene. The results suggest that host cell permissiveness to MHV infection correlates with cellular susceptibility to membrane fusion mediated by the MHV E2 glycoprotein. In addition, we utilized a genetic approach to the analysis of host cell functions involved in determining permissiveness to MHV. By using the chemical mutagen ethyl methanesulphonate, mouse fibroblast cell mutants were generated and selected for their resistance to cell killing by MHV. When challenged with MHV, all five mutants examined gave rise to persistent infections, in contrast to wild-type L-2 cells which were rapidly killed by the virus. The results provide genetic evidence in support of a previous correlation proposed between MHV permissiveness and two host determinants, namely susceptibility to MHV infection and to MHV-mediated cell fusion. Fusion resistance was specific to fusion mediated by the MHV E2 glycoprotein as shown in contact fusion assays between uninfected cells and cells infected either with MHV or with an E2-expressing recombinant vaccinia virus. In contrast, mutant cells were not resistant to fusion after treatment with polyethylene glycol. The observed high rate of generation of these mutants suggests that the conversion of a fully MHV-susceptible cell to a semi-resistant one is a fairly common event, possibly involving a single mutation. In this case, resistance to MHV infection and to E2-mediated membrane fusion may depend on a common host function. This result provides prospects for the precise genetic and biochemical characterization of the steps involved in host cell permissiveness to MHV infection. PMID- 2558161 TI - The site of bluetongue virus attachment to glycophorins from a number of animal erythrocytes. AB - Bluetongue virus (BTV) was shown to agglutinate human, ovine and porcine erythrocytes. Removal of neuraminic acid (NA) from erythrocytes by Vibrio cholerae neuraminidase prevented their agglutination. Haemagglutination was also inhibited by N-acetyl neuraminic acid (NANA), N-glycol neuraminic acid (NGNA) and N-acetyl neuramin-lactose. The ability of BTV to agglutinate trypsin-treated human erythrocytes, which lack the amino-terminal domain and the single N-linked oligosaccharide of glycophorin A, suggests that the virus bound to human erythrocytes via NANA-containing, O-linked oligosaccharides. Glycoproteins with NA-containing oligosaccharides of known structure such as mucin, fetuin, alpha 1 acid glycoprotein, ovomucoid and ovine, porcine, human and equine glycophorin were examined for their ability to inhibit BTV-mediated agglutination of human, ovine and porcine erythrocytes. All glycoproteins containing NANA- or NGNA alpha 2-6GalNAc were capable of inhibiting the agglutination of human and porcine erythrocytes. Treatment of human erythrocytes with Newcastle disease virus neuraminidase and of porcine erythrocytes with Clostridium perfringens neuraminidase to cleave preferentially the NANA- and NGNA alpha 2-3Gal linkages respectively, were shown to have little effect on the ability of the erythrocytes to be agglutinated by BTV. The results suggested that BTV binds to NANA- and NGNA alpha 2-6GalNAc residues in the O-linked oligosaccharides of human and porcine glycophorins respectively and indicated the presence of different binding sites on the virus for erythrocytes from other species. PMID- 2558163 TI - Detection of defective genomes in hepatitis A virus particles present in clinical specimens. AB - Hepatitis A virus (HAV) particles harbouring a physically defective RNA genome have been reported to occur in all HAV-infected cell culture systems analysed so far. The most prominent defects consist of three distinct overlapping deletions in the region of the HAV genome encoding the structural proteins. By probing for the endpoints of these deletions in RNA samples using S1 nuclease and exonuclease VII mapping, we obtained suggestive evidence for the existence also of defective genomes in HAV particles present in faecal specimens, in viraemic blood collected in the course of hepatitis A virus infection in man, as well as in the liver of an experimentally infected marmoset monkey. The deletions identified extend from nucleotide (nt) 1200 to nt 3820 and from nt 1200 to nt 3240 of the HAV genome. They are compatible with two of the deletions detected in particles grown in vitro in cell cultures and shown to interfere with the replication of standard hepatitis A virions. PMID- 2558164 TI - Studies on inhibitors of skin tumor promotion, VI. Inhibitory effects of quinones on Epstein-Barr virus activation. AB - To search for possible antitumor promoters, we carried out a primary screening of fifty-one quinones (anthraquinones, naphthoquinones, azaanthraquinones, and azafluorenones) and related compounds, using their possible inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O tetradecanoylphorbol-13-acetate (TPA) in Raji cells. Some of these quinones, notably 5-hydroxy-1,2-methylenedioxy-anthraquinone [9], shikonin [29], 2 acetylfuranonaphthoquinone [32], 5,8-dihydroxycleistopholine [37], and 5,8 dihydroxy-2-methyl-1-azaanthraquinone [45], were observed to significantly inhibit the EBV-EA activation at low doses. The position and number of hydroxyl groups on phenyl rings of these quinones affected the inhibitory activity on EBV EA activation. The investigation indicates that 9, 29, 32, 37, and 45 might be valuable anti-tumor promoters. PMID- 2558162 TI - Asynchronous mixed infection of Culicoides variipennis with bluetongue virus serotypes 10 and 17. AB - Culicoides variipennis (Diptera: Ceratopogonidae) the primary vector of bluetongue virus (BTV) in the U.S.A. were asynchronously mixedly infected with two BTV serotypes (BTV-10 and BTV-17); flies first ingested a blood meal that contained BTV-17 and 1, 3, 5, 7, and 9 days later selected flies ingested a second blood meal that contained BTV-10. Control flies ingested each parental virus separately, or both viruses simultaneously, in a single blood meal. Electrophoretic analysis of progeny virus clones indicated that superinfection with BTV-10 occurred when the flies ingested the second virus 1, 3 and 5 days post-initial infection. Parental BTV-17 and reassortant virus clones were isolated from these flies, but parental BTV-10 virus was not isolated from any flies. Reassortant clone frequencies were 67%, 71% and 17% when superinfection occurred on days 1, 3 and 5 after initial infection, respectively, as compared to 48% for simultaneously infected flies. Only parental BTV-17 clones were isolated from flies that ingested the second virus on days 7 and 9 after initial BTV-17 infection. The results indicated that interference to superinfection occurred in C. variipennis by 5 days and flies were refractory to superinfection by 7 days post-initial infection. Analysis of segregation of the parental origin of genome segments in the reassortant clones indicated selection against most segments of BTV-10 parental origin. This occurred both in individual flies and in individual groups. The fact that C. variipennis readily fed on a second blood meal and their ability to produce new viral genotypes suggested that these vectors are highly permissive hosts for evolution of BTV by genome reassortment. PMID- 2558165 TI - Chondroid syringoma. A review of the literature and case report. AB - The third documented case of chondroid syringoma occurring in a toe, involving the largest tumor of the three, along with a review of the literature, has been presented. The possibility of a mixed tumor should be kept in mind when considering the differential diagnosis of a solid nodule in the skin, particularly if it is of long duration. These tumors usually will present as simple, asymptomatic, subcutaneous nodules that are shelled out easily during surgical excision. However, there is a tendency toward local recurrence that usually is attributed to incomplete surgical removal, which easily may occur because of the lobulation of the tumor. Complete excision of the primary tumor always is recommended, as diagnosis can be made only by histopathologic examination. Six reported cases revealed that the recurrence of the lesion following primary excision resulted in malignant degeneration. In nine reported cases where metastasis had occurred, the initial site of metastasis in five patients was the regional lymph nodes, and, in four patients, there was metastasis in the lung. The period of time between the removal of the primary lesion and the malignant local recurrence or metastasis has been reported to be from 1 to 4 years. Recognition of chondroid syringoma is clinically difficult because of its rarity. Its diagnosis requires histologic examination. PMID- 2558166 TI - Eccrine poroma. PMID- 2558167 TI - Regulation of N-methyl-D-aspartate receptors revealed by intracellular dialysis of murine neurones in culture. AB - 1. The whole-cell patch clamp recording technique was employed to investigate the intracellular regulation of N-methyl-D-aspartate (NMDA) receptors in cultured murine hippocampal neurones. Excitatory amino acids were repeatedly applied at regular intervals during intracellular dialysis with solutions of various composition. 2. Currents evoked by L-aspartate, an agonist of NMDA receptors, gradually 'washed out' to approximately 50% of their initial amplitude during dialysis with an intracellular solution containing CsCl and EGTA as a calcium buffer. In contrast, responses to kainate did not wash out. The wash-out of L aspartate currents followed an exponential time course with a time constant of about 150 s. Wash-out did not appear to be related to desensitization of NMDA receptors. 3. Following wash-out, L-aspartate responses were blocked by Mg2+, ketamine or D-2-amino-5-phosphonovalerate indicating that these responses were still mediated by NMDA receptors. Furthermore, responses to NMDA itself showed wash-out to the same extent and with a time course similar to that for L aspartate responses. 4. Neither the time course nor the extent of the wash-out of responses to L-aspartate was affected when the Ca2+ concentration of the dialysate was varied from zero to 1.5 x 10(-5) M. In addition, wash-out was unaffected by substitution of BAPTA for EGTA, indicating that wash-out was not a consequence of changes in intracellular pH related to the binding of Ca2+ to the buffer or to the kinetics of this binding. Therefore, the wash-out of NMDA currents could not be attributed to a gradual elevation of the concentration of intracellular Ca2+. 5. The extent of the wash-out of L-aspartate currents was similar for cells held at +40 versus -60 mV although the rate of wash-out was slower at the depolarized potential. In addition, the reversal potential of these currents was not altered, demonstrating that a change in driving force did not account for a component of the wash-out. 6. Inclusion of an ATP regeneration solution (Forscher & Oxford, 1985) in the dialysate prevented the wash-out of L aspartate currents. ATP alone was less effective in preventing wash-out whereas phosphocreatine and creatine phosphokinase were ineffective by themselves. Wash out also occurred when ATP was replaced with the non-hydrolysable analogue, beta, gamma-methyleneATP, or with GTP. In cells where wash-out of L-aspartate currents had been established, subsequent dialysis with the ATP regenerating solution partially reversed this wash-out.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2558168 TI - Voltage-activated membrane currents in rat cerebellar granule neurones. AB - 1. Voltage-activated currents have been recorded from cerebellar granule neurones in explant cultures from young rats (1-9 days old). Cells were examined with whole-cell patch-clamp methods. Depolarizing pulses from a pre-pulse potential of -100 mV evoked a rapidly activated transient inward current, and an outward current which decayed in two phases. The ionic dependence, kinetics and pharmacological properties of these currents have been studied. 2. Peak inward Na+ currents in cells from 7-day-old rats were in the range 350-450 pA. No evidence was found for the presence of calcium currents. Thus, inward current was unchanged in zero Ca2+, 1 mM-EGTA solution. No inward current was obtained in medium containing 10 mM-Ba2+ and tetrodotoxin (TTX). Supplementing the pipette (i.e. intracellular) solution with Mg-ATP did not reveal any Ca2+ current. 3. Depolarizing steps (from -100 mV) in TTX-containing solution gave an early transient outward current and a late outward current. The transient current resembled IA described in other cells, and reversed close to EK in both normal and elevated potassium concentrations, indicating that K+ is the predominant charge carrier. Depolarizing steps from -50 mV failed to give a transient outward current, and gave only a slowly rising current which resembled the late potassium current, IK. 4. Inactivation of the transient current was examined by applying test depolarizations from increasingly negative pre-pulse potentials (-50 to -120 mV): half-inactivation occurred at -72 mV. Transient outward currents decayed exponentially with time constants, tau, of 7.3-25.3 ms at 0 mV. The time course of removal of inactivation in cells held at -50 mV, and given increasingly long pre-pulses to -100 mV, was exponential with tau = 35 ms. 5. Both transient and late outward currents were reversibly abolished by addition to the bathing medium of 10 mM-Ba2+ or 1 mM-quinine. Outward K+ current was not dependent on external calcium. Tetraethylammonium (20 mM) selectively reduced the late outward current; the peak transient current was reduced by less than 20%. 4-Aminopyridine (2 mM) showed little selectivity between transient and late outward currents. 6. It is concluded that cerebellar granule cells from young rats possess voltage-activated inward Na+ current as well as two types of K+ current, IA and IK. In terms of neuronal functioning, the properties of the transient outward current may confer a role in regulating excitability and in repolarization, but a definitive statement will require knowledge of the cellular location and relative densities of channels in granule cells in vivo. PMID- 2558169 TI - Increased sodium pump activity following repetitive stimulation of rat soleus muscles. AB - 1. Soleus muscles of anaesthetized rats were stimulated tetanically (4 s at 20 Hz every 5 s for 5 min), following which the resting and action potentials were measured in surface fibres. 2. At the end of the stimulation period, the mean resting potential was found to have increased from a control value of -79.5 +/- 4.8 mV (mean +/- S.D.) to -90.5 +/- 6.3 mV. The hyperpolarization started to decline after 9 min but was still present at 15 min. 3. Associated with the membrane hyperpolarization was an increase in the mean amplitude of the muscle fibre action potential, from 82.2 +/- 10.8 to 96.8 +/- 10.0 mV. 4. Both the hyperpolarization and the enlargement of the muscle fibre action potential were abolished by 1.25 X 10(-4) M-ouabain, cooling the bathing fluid to 19 degrees C or removing K+ from the bathing fluid. 5. The results are explained in terms of an increase in electrogenic sodium pump activity resulting from tetanic stimulation. When the bathing fluid contained 20 mM-K+, the mean resting potential of stimulated fibres was approximately -30 mV greater than that calculated from the Goldman-Hodgkin-Katz equation. 6. The increase in sodium pumping not only acts to restore the concentrations of Na+ and K+ on either side of the muscle fibre membrane, but, through its electrogenic effect, enables fibres to remain excitable during continuous contractile activity. PMID- 2558170 TI - Modulation of an electrical synapse between solitary pairs of catfish horizontal cells by dopamine and second messengers. AB - 1. Retinas from channel catfish were dissociated and the cells maintained in culture. Horizontal cells that normally receive input from cone photoreceptors were identified. The conductance of the electrical junction formed between a pair of 'cone' horizontal cells was measured by controlling the membrane voltage of each cell with a voltage clamp maintained through either a micropipette or a patch pipette. The two techniques yielded similar results. 2. Transjunctional current was measured while transjunctional voltage was stepped to values between +/- 60 mV. The current (measured 5 ms after a step) was proportional to voltage over the range tested. For steps to voltages greater than +/- 45 mV, the current exhibited a slight time-dependent decline. 3. Dopamine decreased junctional conductance in a dose-dependent fashion. A 50% reduction was obtained with 10 nM dopamine. The D1 agonist fenoldopam (100 nM) also decreased junctional conductance. The uncoupling produced by either agent was rapid and reversible. 4. The introduction of 100 microM-cyclic AMP into one cell of a pair decreased junctional conductance by, on average, 40%. Forskolin (1-10 microM), an activator of adenylate cyclase, decreased junctional conductance 50-90%. 5. The introduction of 80 microM-cyclic GMP into one cell of a pair decreased junctional conductance by, on average, 40%. Nitroprusside (1-10 microM), an activator of guanylate cyclase, reduced junctional conductance 40-65%. 6. The introduction of a peptide inhibitor specific for the cyclic AMP-dependent protein kinase reversed a decrease in junctional conductance produced by superfusion with either dopamine (1 microM), fenoldopam (100 nM) or forskolin (5-10 microM). 7. Intracellular Ca2+ concentration was measured with the fluorescent indicator Fura-2. The intracellular Ca2+ concentration was increased by activation of a Ca2+ current. Junctional conductance remained constant as the internal Ca2+ concentration changed from 100 to 700 nM. 8. Intracellular pH was measured with the fluorescent indicator bis-carboxyethylcarboxyfluorescein. The application of acetate (2.5 mM) reduced intracellular pH by 0.2-0.3 units and decreased junctional conductance by approximately 50%. A subsequent application of fenoldopam did not alter intracellular pH, but decreased junctional conductance by more than 50%. 9. The sensitivity of the junctional conductance between isolated horizontal cells to dopamine is consistent with dopamine having a direct effect on coupling in intact retina. Dopamine regulates the activity of a cyclic AMP-dependent protein kinase which in turn modulates junctional conductance. Changes in intracellular pH and Ca2+ concentration are not involved in mediating the effect of dopamine on coupling. Cyclic GMP and intracellular pH may participate in regulatory pathways independent of that used by cyclic AMP. PMID- 2558171 TI - Dual action (stimulation, inhibition) of D600 on contractility and calcium channels in guinea-pig and cat heart cells. AB - 1. We examined the effects of D600 (0.2-40 microM, generally 2 microM) on the following (i) developed tension in guinea-pig papillary muscles, (ii) calcium current (Ica) and tension in cat ventricular muscle strands, (iii) Ica in guinea pig and cat ventricular myocytes, (iv) single Ca2+ channel currents carried by Ba2+ in cell-attached membrane patches of guinea-pig ventricular myocytes, and (v) Ba2+ currents through dihydropyridine (DHP)-binding sites (skeletal muscle) reconstituted into single functional Ca2+ channels in lipid bilayers. 2. In 27 of 140 preparations studied, D600 elicited a transient stimulation that preceded marked inhibition. The stimulation was normally of short duration (less than 5 min) and moderate strength (less than 50% increase). 3. D600 had no effect on the unit conductance of single cardiac Ca2+ channels. Stimulation was characterized by a decrease in the number of records with no openings (blanks) and an increase in the open-state probability of non-blanks (longer open times, shorter closed times). Inhibition began with an increase in the number of blanks and later included a curtailment of open times and a prolongation of closed times. The net effect after 9 min D600 was a 75% reduction in average current amplitude. 4. A similar pattern of changes in channel open and closed times produced enhancement and then depression of time-averaged open-state probability in single reconstituted channels. 5. Single Ca2+ channel current that was stimulated by adrenaline was only slightly depressed after 2 microM-D600 for 30 min. It may be that channel phosphorylation or Gs-protein activation following beta-receptor stimulation reduces channel affinity for D600. 6. Short-lived binding of D600 to a single inhibitory site may enhance association/activation of Gs-protein and thereby cause transient up-regulation prior to increased drug occupancy and inhibition. Alternatively, there may be separate stimulatory and inhibitory sites. One aspect of inhibition, the increased frequency of blanks, is attributed to a stabilization of the inactivated state; the other aspect, changes in fast kinetics, seems to require a different explanation. PMID- 2558172 TI - Intrinsic properties of nucleus reticularis thalami neurones of the rat studied in vitro. AB - 1. Neurones of the nucleus reticularis thalami of the rat were studied by intracellular recordings from in vitro slices. The resting membrane potential was -56.28 +/- 5.86 mV (mean value +/- S.D.); input resistance was 43.09 +/- 9.74 M omega; the time constant tau was 16.51 +/- 3.99 ms. At the resting membrane potential tonic firing is present, while at membrane potentials more negative than -60 mV a burst firing mode gradually prevails. 2. Prolonged depolarizing current pulses superimposed on a steady hyperpolarization consistently activated sequences of burst-after-hyperpolarization complexes. The all-or-none burst response consisted of Na+-mediated, TTX-sensitive fast action potentials superimposed on a low threshold spike (LTS). The burst was followed by a stereotyped after-hyperpolarization lasting 100-120 ms (BAHP), with a maxima -85 mV. The BAHP was blocked by Cd2+ and apamine but not by 8-Br cyclic AMP. The early component of BAHP was significantly attenuated by TEA. The oscillatory rhythmic discharges were abolished by agents which blocked the BAHP. 3. The presence of strong after-hyperpolarizing potentials (SAHP and BAHP) in RTN neurones plays a significant role in determining two different functional states, defined as tonic and oscillatory burst firing modes, respectively. PMID- 2558173 TI - Low-voltage-activated calcium current in rat aorta smooth muscle cells in primary culture. AB - 1. Electrical and pharmacological properties of the low-voltage-activated Ca2+ current (ICa, LVA) in rat aorta smooth muscle cells (SMC) in primary culture were examined, particularly in comparison with the high-voltage-activated Ca2+ current (ICa, HVA). Both types of Ca2+ currents were recorded in external solution containing 20 mM-Ca2+, using the whole-cell voltage-clamp technique. 2. ICa, LVA was evoked by step depolarizations to potentials more positive than -60 mV from a holding potential of -100 mV, and reached a peak in the current-voltage (I-V) relationship around -30 mV. ICa, HVA was activated at -20 mV, and reached a peak at +20 mV. 3. The intracellular dialysis of 5 mM-F- irreversibly suppressed ICa, HVA, with time, while it has little effect on the ICa, LVA. The ICa, LVA could be separated from the ICa, HVA by either selecting the holding and test potential levels or by perfusing intracellularly with F-. 4. The ratio of peak amplitude of Ba2+, Sr2+ and Ca2+ currents in the respective I-V relationship was 1.6:1.2:1.0 for high-voltage-activated Ca2+ channels and was 1.0:1.4:1.0 for low-voltage activated ones. 5. The inactivation phase of ICa, HVA was fitted by a sum of double-exponential functions, the time constants of which were larger when the current was carried by Ba2+ than by Ca2+. The inactivation time course of ICa, LVA was fitted by a single-exponential function, and the time constant was practically the same when the current was carried by Ba2+ or by Ca2+. Activation and inactivation processes of ICa, LVA were potential-dependent. 6. The steady state inactivation curve of ICa, LVA was fitted by the Boltzmann equation, having a mid-potential of -80 mV and a slope factor of 5.0. The recovery time course from steady-state inactivation was fitted by a sum of two exponential functions. The time constants of the faster phase were 230 and 380 ms, and those of slower phase were 2.8 and 1.8 s at the repolarization potentials of -120 and -100 mV, respectively. 7. The amplitude of ICa, LVA depended on the external Ca2+ concentration ([Ca2+]o), approaching saturation at 95 mM [Ca2+]o. 8. Various polyvalent cations blocked both types of Ca2+ current reversibly in the order (IC50 in M): La3+ (8 x 10(-8)) greater than Cd2+ (6 x 10(-6)) greater than Ni2+ (1 x 10(-5)) greater than Zn2+ (2 x 10(-5)) for ICa, HVA, and La3+ (6 x 10(-7)) greater than Zn2+ (3 x 10(-5)) greater than Cd2+ (4 x 10(-4)) greater than Ni2+ (6 x 10(-4)) for ICa, LVA.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2558175 TI - Electromyographic cross-talk within a compartmentalized muscle of the cat. AB - 1. Experiments were conducted to test the extent to which the electromyographic (EMG) activity generated by the activation of single motor units is conducted from one neuromuscular compartment of the cat lateral gastrocnemius (LG) muscle into adjacent compartments. 2. Potentials produced by stimulation of forty-five single motor units were monitored from bipolar fine-wire EMG electrodes which had been implanted either into the centres of each of the four neuromuscular compartments of LG or into regions of the muscle known to lie on the border of contiguous compartments. 3. In all cases single unit potentials could be recorded from the electrodes in the centre of the compartments which clearly identified the compartment of residence of the muscle unit. Regardless of unit type, the amplitude of the potential recorded from electrodes in one compartment was always greater than that recorded from any other compartment. 4. Smaller potentials could be recorded from electrodes in the centre of compartments adjacent to the compartment of residence of the muscle unit. For those motor units where the amplitude of the EMG potentials recorded from the compartment of residence was large, the amplitude of such 'cross-talk' could be greater than the amplitude of potentials recorded from the compartment of residence of smaller motor units. 5. In the case of electrodes placed at compartment boundaries, no clear compartment selectivity of recording of motor unit potentials was evident. 6. These results indicate that great care must be taken in choosing sites of EMG electrode placement when performing kinesiological studies, especially when the amplitude of the EMG activity recorded is of consideration. PMID- 2558174 TI - Desensitization of swine arterial smooth muscle to transplasmalemmal Ca2+ influx. AB - 1. The effect of transplasmalemmal Ca2+ influx on the [Ca2+]i dependence of smooth muscle contraction was evaluated by measuring intracellular [Ca2+] (as estimated by aequorin), myosin phosphorylation, and isometric stress in swine carotid media. 2. Extracellular Ca2+ was removed by incubation in physiological saline with 1 mM-EGTA and no added CaCl2 for 20 min (termed EGTA treatment). In some preparations, intracellular Ca2+ was released by a brief (5 min) histamine stimulation while in this Ca2(+)-free EGTA solution (termed histamine treatment). 3. Restoration of extracellular CaCl2 to EGTA and histamine-treated preparations in the continued presence of histamine was associated with an initial large aequorin light transient. However, this light transient was not initially associated with an increase in myosin phosphorylation or rapid stress development, suggesting that the contractile apparatus was desensitized to aequorin-estimated myoplasmic [Ca2+]. The desensitization was temporary, and resolved by 10 min after restoration of extracellular CaCl2. 4. The light transient observed upon restoration of extracellular CaCl2 was smaller in preparations only EGTA treated when compared to preparations treated with both EGTA and histamine, suggesting that histamine treatment further desensitized the contractile apparatus. 5. The stress development rate was not slowed when histamine and extracellular CaCl2 were simultaneously added to EGTA-treated preparations, suggesting that the desensitization was only to transplasmalemmal Ca2+ influx (from extracellular CaCl2 readdition), and not intracellular Ca2+ release (from the histamine stimulation). 6. In EGTA and histamine-treated preparations, restoration of extracellular CaCl2 in the presence of 109 mM-KCl was associated with a larger aequorin light signal than was observed upon readdition of CaCl2 in the presence of histamine, suggesting that depolarization also further desensitized the contractile apparatus. 7. Depolarization of EGTA treated preparations did not increase [Ca2+] or stress, suggesting that depolarization did not release intracellular Ca2+ stores. 8. No significant light transient was observed upon addition of extracellular LaCl3, suggesting that tissue damage or leakage of aequorin into the extracellular space was not the cause of the Ca2(+)-reintroduction light signal. 9. These data suggest that removal of extracellular CaCl2 desensitizes the contractile apparatus of smooth muscle to transplasmalemmal Ca2+ influx. This desensitization is only to readdition of extracellular Ca2+; the contractile apparatus still responds to intracellular Ca2+ release. The desensitization is increased by prior depolarization or brief histamine treatment (potentially by depleting intracellular Ca2+). The source of activator Ca2+ appears to affect the relationship between aequorin light and phosphorylation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2558176 TI - Effects of hypoxia and metabolic inhibition on the intracellular sodium activity of mammalian ventricular muscle. AB - 1. Intracellular Na+ activity (aiNa) has been measured in Purkinje fibres from sheep heart and in ventricular muscle from rabbit heart during hypoxia and metabolic inhibition. The aiNa was measured using liquid sensor ion-sensitive microelectrodes. 2. Hypoxia, produced by replacement of O2 with N2 in the superfusate, produced an increase in aiNa. This increase was larger if sucrose replaced glucose in the superfusing Tyrode solution. The increase in aiNa was accompanied by a small depolarization. Upon reoxygenation aiNa decreased and cells rapidly repolarized. 3. When oxidative phosphorylation was inhibited by application of 2 mM-cyanide, aiNa increased. This increase was also accompanied by a small depolarization. Upon removal of cyanide, aiNa and membrane potential recovered to control levels. 4. After inhibiting glycolysis, by replacing glucose with 2-deoxy-D-glucose, inhibition of oxidative phosphorylation (by addition of cyanide or exposure to hypoxia) produced a much more rapid increase in aiNa and a large contracture. The rise in aiNa and the occurrence of a contracture could not be inhibited by application of amiloride (1 mM) or tetrodotoxin (1 microgram ml 1). Removal of cyanide or reoxygenation and replacement of glucose resulted in a rapid relaxation of the contracture and a slower decrease in aiNa. 5. The relative rates of increase in aiNa during metabolic inhibition were compared with the rate observed when Na+-K+-ATPase was inhibited by application of 10 mumols l 1 of the cardio-active steroid strophanthidin. The rate of increase of aiNa when both oxidative phosphorylation and glycolysis were inhibited was approximately twice that observed with only oxidative phosphorylation inhibited and approximately half that observed in the presence of 10 microM-strophanthidin. 6. Cyanide, applied when aiNa had been elevated (i.e. during exposure to 10 microM strophanthidin to inhibit Na+-K+-ATPase), did not produce a contracture. The contracture observed in the presence of cyanide and 2-deoxy-D-glucose still occurred when Ca2+ was removed from the superfusate. PMID- 2558177 TI - Effects of defolliculation on membrane current responses of Xenopus oocytes. AB - 1. Catecholamines, adenosine, gonadotrophins, vasoactive intestinal peptide (VIP) and E-series prostaglandins all elicit K+ currents in follicle-enclosed Xenopus oocytes. Evidence suggests that cyclic nucleotides act as intracellular messengers in the activation of this K+ conductance. Muscarinic agonists and some divalent cations (e.g. Co2+, Mn2+, Ni2+ and Cd2+) elicit slow oscillatory Cl- currents, which are activated through hydrolysis of inositol phospholipids and mobilization of intracellular calcium by inositol phosphates. 2. We investigated whether these membrane current responses were generated in the oocyte itself or in enveloping follicular cells which are coupled to the oocyte by gap junctions. Oocytes were defolliculated, either enzymatically using collagenase, or by manual dissection combined with rolling over poly-L-lysine-coated slides. Removal of follicular cells was checked using scanning electron microscopy. Membrane current responses of defolliculated oocytes were compared with responses seen in follicle enclosed oocytes taken from the same ovary. 3. The K+ responses evoked by all the various hormones/neurotransmitters were either drastically reduced (greater than 90%) or abolished by defolliculation. K+ currents generated by the adenylate cyclase activator forskolin and by intraoocyte injection of adenosine 3',5' cyclic monophosphate (cyclic AMP), or guanosine 3',5'-cyclic monophosphate were similarly reduced in defolliculated oocytes. In contrast, oscillatory Cl- currents to acetylcholine and divalent cations were selectively preserved through defolliculation. 4. Injection of cyclic AMP (1-20 pmol) into defolliculated oocytes had little or no effect on oscillatory Cl- currents elicited by ACh. However, the calcium-dependent transient Cl- current, activated by depolarization of the oocyte membrane, was consistently potentiated (100-900%) by injections of cyclic AMP (1-10 pmol). 5. These experiments suggest that cyclic nucleotide activated K+ currents arise essentially in follicular cells and are monitored within the oocyte through electrical coupling by gap junctions. Oscillatory Cl- responses evoked by ACh and divalent cations are produced largely or wholly in the oocyte itself. PMID- 2558179 TI - Explants of human oral epithelium exposed to viruses and cancer chemotherapeutics. AB - Cultures of proliferating epithelial cells were established from explants of normal human oral epithelium from healthy young volunteers. The epithelial cells were found permissive for herpes simplex virus type 1 and type 2, coxsackie virus A-4 and A-16, adenovirus type 5, measles vaccine, rubella and influenza type A virus-. Medium from DEAE-pretreated epithelial cultures infected with two subtypes of human immunodeficiency virus-1 showed an increasing content of virusprotein with time by antigen ELISA testing. In contrast there was no evidence of infection with coxsackie virus type B-2, cytomegalovirus, Epstein Barr virus and varicella zoster virus. Treatment of the epithelial cells with a non-cytotoxic dose of cancer chemotherapeutic prior to or after infection with coxsackie virus A-4 or herpes simplex virus type 1 influenced the virus production dependent on both compound, mode of application, and virus. Adriamycin (doxorubicin) in low dose was found to stimulate the production of the two viruses. PMID- 2558178 TI - Heterogeneous distribution of fast and slow potassium channels in myelinated rat nerve fibres. AB - 1. Potassium currents were measured in voltage-clamped single myelinated rat nerve fibres before and after paranodal demyelination with 0.2% pronase or 0.2% lysolecithin added to the external solution. Sodium currents were blocked by 300 nM-tetrodotoxin. For the purpose of comparison, intact frog nerve fibres were also investigated. 2. Our results suggest the existence of at least two distinct types of K+ channels in the intact node of Ranvier, one with slow and another with fast gating kinetics, in the ratio 4:1. 3. In the rat nodal membrane, slow K+ channels have voltage-dependent time constants of K+ deactivation with tau n = 68 ms at E = -105 mV and tau n = 26 ms at E = -150 mV at 20 degrees C. The activation curve of the slow K+ conductance is sigmoid with an inflexion point at -60 mV. This means that about 35% of the slow K+ channels are in the open state at the resting potential of -77 mV. Slow K+ channels could be blocked by 10 mM tetraethylammonium chloride, but were insensitive to 4-aminopyridine. 4. After paranodal demyelination the ratio of fast to slow K+ channels increased from 17 to 83%. As in the frog (Dubois, 1981 alpha), the population of fast K+ channels in the rat may consist of two different subgroups, both of which can be blocked by 4-aminopyridine. 5. Demyelination was accompanied by an increase in the capacity current which was used to estimate the exposed membrane area. The density of slow and fast K+ channels was calculated from the quotient of the steady-state K+ conductance to membrane area. The density of the slow K+ channels is maximal in the nodal membrane and decreases to 1/31 in the internode. By contrast, the distribution of the fast K+ channels differs, their density being maximal in the paranode and decreasing to one-sixth in the node and internode. PMID- 2558180 TI - [The gating mechanism of K(+)-channels coupled to the FMRFamide receptor in the ganglion cells of Aplysia]. AB - 1. Neurons with a receptor responded to FMRFamide (Phe-Met-Arg-Phe-NH2) were identified in the ganglion of Aplysia kurodai. Ionic mechanism and channel gating system of the FMRFamide-induced responses were investigated by current clamp and voltage clamp methods. 2. The reversal potential of FMRFamide-induced response exactly coincided with the equilibrium potential for K+. This proved that the response was produced by a specific increase in membrane permeability toward K+, exclusively. 3. The FMRFamide-induced response was not affected by the inhibitors for Ca2(+)-activated K(+)-current, i.e., TEA, apamin, and EGTA. This excluded a possibility that FMRFamide-activated K(+)-channel is a Ca2(+)-activated K(+) channel. 4. Intracellular injection of pertussis-toxin (PTX) caused no change in either resting potential or conductance, but it irreversibly blocked the FMRFamide-induced outward current within 30 min. Similarly applied cholera toxin (CTX) showed no effect on the FMRF-amide response. 5. Intracellular application of guanosine 5'-0-(2-thiodiphosphate) (GDP beta S) caused no effect on either resting potential or conductance, but it blocked the FMRFamide-induced K(+) current within 3 min. 6. Intracellular application of guanosine 5'-0-(3 thiotriphosphate) (GTP gamma S) alone induced a slowly developing, irreversible outward current associated with an increase in membrane conductance. However, repetitive applications of FMRFamide immediately after the start of GTP gamma S application markedly facilitated the effect of GTP gamma S on the resting membrane. 7. Intracellular application of either adenylate cyclase inhibitor (3' deoxyadenosine) or A-kinase inhibitor (H-8) did not affect the FMRFamide-induced response. 8. It was concluded that the FMRFamide-induced K(+)-current is mediated by PTX-sensitive GTP-binding protein Gi, Go or Gk. It was also suggested that the FMRFamide-induced response is produced independently of the changes in intracellular Ca2+ or cyclic AMP. PMID- 2558181 TI - Rehabilitation of a patient with a bilateral partial maxillary resection. A clinical report. PMID- 2558182 TI - Is yawning an arousal defense reflex? AB - Despite the fact that yawning is a reality of everyday life, its study is not included in the curriculum of medical schools, and most medical textbooks barely mention its existence. Two factors may help to explain this puzzling situation: (a) yawning's borderline position between psychology and neurology, and (b) researchers' lack of understanding as to why people yawn. After review of the literature and personal observation, it is concluded that yawning is a complex arousal defense reflex located in the reticular brainstem with a peripheral and central arche, whose aim is to reverse brain hypoxia. Yawning occurs with loss of interest (boredom) and may or may not be associated with fatigue. By reversing drowsiness, yawning avoids a decreased concentration capacity resulting from borderline hypoxia. It is hoped that this article will stimulate further research on the phenomenon. PMID- 2558183 TI - Practice budgets: lifting the veil of ignorance. PMID- 2558184 TI - Classification of psychosocial disturbance in general practice. PMID- 2558185 TI - Survey to establish the incidence of minor side effects in infants following protective immunization. AB - A study of the minor side effects from the immunization of children against diphtheria/pertussis/tetanus, diphtheria/tetanus, and measles is reported. The sample of 306 children received 1028 vaccinations. A secondary study of measles vaccine was made on 177 immunizations. A diary card was used to provide control data before injections and to measure the increase in incidence of minor symptoms after injection. The reported incidence of side effects after both diphtheria/tetanus and measles vaccinations was low and the patterns similar. The increase in side effects was greater after diphtheria/pertussis/tetanus injection, particularly when there was soreness at the injection site. The incidence of soreness was lower when the site of the injection was the buttock rather than the arm. The diary card was found to be an effective method of providing control data and of monitoring any increase in the incidence of minor symptoms following immunizations. The information obtained should assist health care workers to provide accurate advice and to reassure parents who are concerned about their children's protection. PMID- 2558187 TI - Evaluation of a practice-based programme of health checks: financial cost and success at risk detection. AB - With the help and supervision of a facilitator, a scheme of nurse-run health checks was established in 1983 in a suburban group practice of approximately 12,000 patients. The practice receptionists recruited patients opportunistically when they presented to consult a doctor. After two and a half years, 1382 patients had been screened for cardiovascular risk factors; this was 94.0% of those invited to attend and 24.9% of the target population of patients aged 25-64 years. Recording of risk factors increased greatly between 1983 and 1986, although uptake of immunizations was low. Sixty five previously undiagnosed hypertensives were identified, as were 11 new diabetics. This was achieved with two sessions per week; thus to succeed in screening the whole population within five years, four weekly sessions would have been necessary. When the cost of equipment and staff salaries were offset against 70% reimbursement of salaries and item of service fees the net financial cost to practice from 1983-86 was approximately 1.73 pounds per patient, or 477.00 pounds per clinic session per year. PMID- 2558186 TI - Pyridoxine (vitamin B6) and the premenstrual syndrome: a randomized crossover trial. AB - A randomized double-blind crossover trial was conducted to study the effects of pyridoxine (vitamin B6) at a dose of 50 mg per day on symptoms characteristic of the premenstrual syndrome. Sixty three women aged 18-49 years, identified by means of a general practice based survey of menstrual patterns in the community, entered the trial. All of the women had noticed moderate to severe premenstrual symptoms during the previous year. The women kept a daily menstrual diary which graded the severity of nine individual symptoms from zero to three. After completing a diary for an initial month the women were randomized to receive either drug or placebo for three months, after which the treatments were crossed over for a further three months. Thirty two women completed the full seven months of the study. In these women a significant beneficial effect (P less than 0.05) of pyridoxine was observed on emotional type symptoms (depression, irritability and tiredness). No significant effect was observed on premenstrual symptoms of any other type. PMID- 2558189 TI - Study of written communication between general practitioners and a community physiotherapist. AB - Previous studies have shown that written communication between hospitals and general practitioners could be improved. This study investigated whether communication problems exist between general practitioners and a community physiotherapist in primary care. Deficiencies were evident in the written referral information received by the physiotherapist. The general practitioners felt they would like to receive feedback information from the physiotherapist after treatment. The information needs of the general practitioners and community physiotherapist were only partly met, and possible improvements are discussed. PMID- 2558188 TI - Human immunodeficiency virus in drug misusers and increased consultation in general practice. AB - The use of general practitioner services by a group of intravenous drug users was recorded over two two-year time periods 1984-85 and 1986-87. This was felt to represent the period of maximum change in awareness of human immunodeficiency virus (HIV) infection by patients and medical staff. Fifty patients were randomly selected: 25 who were HIV positive and 25 who were HIV negative. Between the two time periods a dramatic increase in consultation rate for both high risk and infected patients attending their general practitioner was recorded (318% and 172% increase, respectively). A small increase in attendance at the accident and emergency department (30% and 34% increase, respectively) was recorded for high risk and infected patients, and there was a large increase in attendance at the infectious diseases unit for infected patients but there was little effect on use of other hospital services. The implications for resource needs in the community are discussed. PMID- 2558191 TI - Enterobius vermicularis: a possible cause of intestinal colic? PMID- 2558190 TI - The exceptional potential of the consultation revisited. AB - A new framework is proposed for analysing the consultation in primary care, and for integrating other models relevant to the consultation. Use of communication skills helps to reconcile the respective agendas of doctor and patient into a negotiated plan, which includes both management of problems and health promotion. Achieving the potential of consultations depends on cooperation between patient and doctor, and on sufficient time for appointments. PMID- 2558192 TI - Effectiveness of anti-smoking advice from doctors who smoke. PMID- 2558193 TI - 'Patient care' and patient benefit. PMID- 2558194 TI - Breast self examination. PMID- 2558195 TI - Screening and the new contract. PMID- 2558196 TI - Screening: the case against. PMID- 2558197 TI - Effect of small group education on the outcome of chronic asthma. PMID- 2558198 TI - GPs' use of hospital investigative facilities. PMID- 2558199 TI - Working for patients. PMID- 2558200 TI - The efficient use of time in general practice. PMID- 2558201 TI - Chronic non-malignant pain: time to take on the challenge. PMID- 2558202 TI - Study of the effect of time availability on the consultation. AB - This study looked at the effect of different appointment time intervals on process and outcome measures in the consultation. Over a five-month period patients attending a two-partner surgery were non-systematically allocated to appointments at five, 10 or 15 minute intervals. Consultations were audiotaped and analysed. When appointments were scheduled at longer intervals, doctors asked significantly more questions and made significantly more statements explaining the problem and its management, while patients asked significantly more questions and made significantly more statements of their own ideas about the problem. In consultations booked at shorter intervals patients were significantly more likely to report in satisfaction questionnaires that they had little or far too little time available. The implications of the results for future planning are discussed. PMID- 2558203 TI - Mathematical relationship between waiting times and appointment interval for doctor and patients. AB - Appointment systems try to achieve a balance between the time the doctor waits for patients to arrive and the time patients spend waiting to be seen. Mathematical analysis reveals that the time a patient can expect to wait increases exponentially as the appointment interval is reduced. An appointment interval that is less than the median consultation length can result in long waits for patients with no saving of time for the doctor. More frequent, shorter surgeries can save time for patients with no increase in the doctor's waiting time. Methods of improving the efficiency of use of surgery time are discussed. PMID- 2558204 TI - Factors affecting the decision to consult with dyspepsia: comparison of consulters and non-consulters. AB - To identify factors associated with the decision to consult with dyspepsia, patients with dyspepsia were identified from a postal survey in the community. A random sample of 69 patients who had consulted their general practitioner and 66 patients with dyspepsia who had not consulted were interviewed in their homes. Differences in consultation behaviour were not explained by differences in self reported severity or frequency of symptoms or by the presence of associated symptoms. The most striking difference between the two groups was concern among the consulters about the possible seriousness of symptoms. Consulters were also more likely to be worried about cancer and heart disease and to have experienced more disruptive or threatening life events than the non-consulters. These results emphasize the importance of looking beyond the presentation of common symptoms in general practice to patients' fears about the significance of the symptoms and to non-physical determinants of consultation behaviour. PMID- 2558205 TI - How well do general practitioners manage dyspepsia? AB - This paper reports the incidence of dyspepsia in general practice, the characteristics of patients, the types of complaints presented and the management of the dyspeptic patient by general practitioners. Fourteen general practitioners in the Maastricht region of the Netherlands studied 318 consecutive patients presenting with dyspepsia. Two questionnaires were used: one filled in by the patient (82% response), the other by the physician (100% response). The diagnostic conclusions which were established after three months of follow-up were compared with the diagnostic hypotheses at the initial consultation. The annual consultation rate for dyspepsia was calculated as 27 per 1000 registered subjects. One third of the patients had an earlier history of dyspepsia. Almost all patients (95%) complained of pain, and 37% had been suffering from pain for more than three months before consulting the general practitioner. The general practitioner prescribed medication in 70% of cases; less commonly the patient was referred for x-ray (14%), endoscopy (13%) or to a specialist (11%). A higher age was associated with a higher probability of referral, and with the finding of organic disease. A history of ulcer disease was strongly correlated with the diagnosis of an ulcer during the current episode. The overall concordance between the general practitioner's diagnostic hypothesis at the initial consultation and the diagnostic conclusion after three months of follow-up was 78%; it was highest when minor pathology was suspected. We conclude that dyspepsia is managed well in general practice and is only rarely associated with major lesions. Dyspeptic patients referred to a specialist therefore constitute a highly selected population. PMID- 2558207 TI - A diagnostic centre for general practitioners: results of individual feedback on diagnostic actions. AB - A diagnostic centre, managing diagnostic tests for general practice, can improve the service provided by primary health care and the communication between general practitioners and specialists. In addition, it can evaluate the use and misuse of tests. This paper describes the work of a diagnostic centre in the Netherlands serving 80 general practitioners. Following the introduction of individual feedback to general practitioners on their use of diagnostic tests there was a decrease in the number of requests for tests. PMID- 2558206 TI - Survey of the use of homeopathic medicine in the UK health system. AB - An analysis of 7218 consultations showed that homeopathic medicines are being used to treat a wide range of morbidity in the United Kingdom. The data were derived from all consecutive consultations during one week by 73 doctors who used homeopathic medicine. Of these consultations 88% were conducted as part of the National Health Service (the majority in general practice). Thirty five per cent overall and 25% of general practice consultations were managed using homeopathic medicines, and these were combined with conventional drugs in 8.5% of the prescriptions. PMID- 2558208 TI - Estimates of general practitioner workload: a review. AB - This paper reviews four studies sponsored by the Department of Health which have attempted to measure workload in general practice and compares these with data from the general household survey. Despite the considerable differences in the objectives and methods employed by the four studies, they were found to contain remarkably consistent measurements of general practitioner workload. In a 'normal working week' general practitioners spend 38 hours on general medical service duties (including 24 hours of patient contact and five hours of travel to home visits), they see 150 patients or their representatives in surgery, and make 26 home visits. In an 'annual average week', taking into account holidays and sick leave, general practitioners undertake 90% of this workload. The studies show consistently large variations in the workload of general practitioners measured in this way, but fail to identify the key determinants of such variations. The reasons underlying the variation in general practitioner workload will remain unclear until we can distinguish between the expected, measurable variation and the residual, unexplained variation which may be due to the personal preferences of general practitioners. PMID- 2558209 TI - Liaison psychiatry in general practice: a comparison of the liaison-attachment scheme and shifted outpatient clinic models. AB - Most psychiatrists who visit health centres use the shifted outpatient clinic model, the main aim of which is to improve secondary care by providing it in the primary care setting. For five years we have employed a liaison-attachment scheme in which support and advice from the psychiatrist enables general practitioners to improve their care of patients with psychiatric and psychological problems. One of the advantages of the latter model is that the psychiatrist can contribute to the care of patients not seen by the specialist psychiatric service and also to the development of the primary care team. The scheme is cost effective as psychiatrists can advise on the care of far more patients than they could see in formal referrals, fewer patients are taken on for a course of psychiatric treatment that could be provided by general practitioners and the skills of general practitioners and their trainees are enhanced. It is hoped that more general practitioners will adopt this pattern of working so that it can be fully developed and evaluated. PMID- 2558210 TI - Snowballing hypothermia alert system or rural noseyness? PMID- 2558211 TI - Classification of psychosocial disturbances in general practice. PMID- 2558212 TI - Adverse effects of screening. PMID- 2558213 TI - Treatment of asthma. PMID- 2558214 TI - Audit projects for medical students. PMID- 2558215 TI - Core data for practice annual reports. PMID- 2558216 TI - The exceptional potential of the consultation revisited. PMID- 2558217 TI - Continuing medical education. PMID- 2558218 TI - Torture of prisoners. PMID- 2558219 TI - New contract. PMID- 2558220 TI - Future of the College. PMID- 2558221 TI - Plasma concentrations of pituitary hormones in 2,3,7,8-tetrachlorodibenzo-p dioxin-treated male rats. AB - Experiments were conducted to test the hypothesis that acute TCDD toxicity is associated with pituitary hypofunction. Sexually mature male Sprague-Dawley rats were given graded doses of TCDD (0-100 micrograms/kg) and evaluated 7 days later. Despite pronounced hypophagia and body weight loss, plasma concentrations of growth hormone (GH), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) were not significantly affected by any dose of TCDD. Only prolactin (PRL) concentrations were reduced, while, as previously reported, thyroid-stimulating hormone concentrations were elevated. Also, plasma LH, PRL, and adrenocorticotropic hormone (ACTH) concentrations were not significantly affected 1, 2, 3, 4, 5, or 7 days after a single dose of TCDD (50 micrograms/kg). We conclude that (1) pituitary hypofunction is not a major cause of the initial stages of acute TCDD toxicity, (2) growth retardation in TCDD-treated rats is not the result of a deficiency of GH, (3) alterations in plasma corticosterone concentrations are due to altered responsiveness of the adrenal to ACTH stimulation rather than to changes in plasma ACTH concentrations, and (4) that impaired spermatogenesis is not associated with a decrease in plasma FSH concentrations. In addition, the lack of a consistent effect on plasma PRL concentrations suggests that alterations in plasma PRL concentrations do not play a critical role in the toxicity of TCDD. Finally, because TCDD treatment causes a serious androgenic deficiency without increasing the rates at which androgens are catabolized or excreted, the fact that plasma LH concentrations were unaffected indicates that TCDD treatment must reduce the responsiveness of the testis to LH stimulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558222 TI - Adenosine production and energy metabolism in ischaemic and metabolically stimulated rat heart. AB - Adenosine may modulate blood flow and electrical activity in heart in response to changes in myocardial energy metabolism. In the present study, 31P NMR spectroscopy was used to examine the relation between cytosolic phosphate metabolite levels and release of adenosine into the venous effluent of isovolumic heart during graded low-flow ischaemia or metabolic stimulation with isoproterenol. When coronary flow rate was varied in steps between 1.6 and 12 ml/min/g, cytosolic ATP levels did not change significantly but the phosphorylation potential exhibited a linear correlation with flow rate below approximately 7 ml/min/g. Purine release (adenosine and inosine) correlated linearly with the cytosolic phosphorylation potential and free AMP concentration. Metabolic stimulation of hearts with isoproterenol (0.4, 3.0, and 60 nM), produced a significant fall in cytosolic ATP levels and decreased the cytosolic phosphorylation potential. Purine release in these hearts increased exponentially as the cytosolic phosphorylation potential dropped, and as cytosolic free AMP increased. These results support a link between the phosphorylation potential and the mechanism of adenosine production during ischaemia and metabolic stimulation. Presumably, this link is the activity of the enzyme 5'-nucleotidase, which is responsible for converting AMP to adenosine, together with the concentration of its substrate, AMP. In low-flow ischaemia, cytosolic AMP may control adenosine formation. With isoproterenol stimulation, a more complex relationship exists, indicating possible allosteric regulation of the enzyme(s) responsible for adenosine formation, in addition to changes in AMP concentration. PMID- 2558223 TI - Oxidative injury to myocardial membrane: direct modulation by endogenous alpha tocopherol. AB - Peroxidation of myocardial-membrane phospholipid is considered an important pathogenic component of heart muscle damage in ischemia and reperfusion. The extent to which membrane alpha-tocopherol (vitamin E) in the heart can modulate such damage and protect against it is a matter of controversy. The relative alpha tocopherol deficit of spontaneously-hypertensive (SH) rat myocardium as compared to the myocardium of the Wistar-Kyoto (W/K) normotensive parent strain prompted use of these animals to identify and characterize any protective antiperoxidant role of endogenous, myocardial-membrane alpha-tocopherol. With exposure to a superoxide- and iron-containing initiator of peroxidation, the membrane complements from the ventricular myocardia of the SH rat and the W/K parent strain were found to have very different peroxidative-injury profiles. SH-rat myocardial membrane demonstrated a marked sensitivity to peroxidation as reflected in the acute onset and rapid progression of phospholipid damage. The greater susceptibility of SH-rat myocardial membrane to free-radical attack could not be explained by inter-strain compositional differences in membrane polyunsaturated fatty acids or fatty aldehydes. Rather, the basis for the enhanced peroxidation was identified as the 3-fold lower alpha-tocopherol content of SH-rat myocardial membrane with respect to the heart-muscle membrane from the normotensive animal. The relative alpha-tocopherol deficit not only increased the susceptibility of SH-rat cardiac membrane to damage under pro-oxidant conditions, but also reduced the efficacy of exogenously supplied antioxidant intervention. These findings demonstrate that membrane alpha-tocopherol tone is a critical protectant of myocardial phospholipid against oxidative injury and acts as a determinant of the course of heart-membrane peroxidative damage. PMID- 2558224 TI - Developmental changes of sarcolemmal Na+-H+ exchange. AB - We previously demonstrated that the effect of respiratory acidosis on cardiac contractility in the newborn was less than in the adult rabbit, and these data suggested a higher [Na+]i and [Na+]i-[Ca2+]o exchange in the newborn as compared to the adult. In this study, we investigated developmental changes of Na+-H+ exchange in isolated sarcolemmal vesicles. Sarcolemmal purification for Na+-K ATPase was 61.9 and 67.1 fold in the newborn and the adult rabbit heart, respectively. In the presence of an outwardly directed proton gradient across the vesicular membrane, sarcolemmal 22Na uptake rate in the newborn (0.22 +/- 0.01 nmol Na+/mg prot/s) was significantly higher than than in the adult (0.16 +/- 0.01 nmol Na+/mg prot/s). 1.0 mM amiloride inhibited 22Na uptake by 75% and 80% in the newborn and the adult, respectively. In the absence of a pH gradient, vesicular 22Na uptake in the newborn and the adult were not significantly different. In conclusion, the higher Na+-H+ exchange in the newborn may lead to a higher [Na+]i and subsequent calcium influx via Na+-Ca2+ exchange as compared with the adult during acidosis. This may explain the greater recovery of mechanical function in the newborn heart as compared to the adult heart during acidosis. PMID- 2558225 TI - Serological survey of antibodies to cytomegalovirus in the Santa Cruz region of Bolivia. AB - Cytomegalovirus (CMV) infection is known to be worldwide, and prevalent in developing countries (Krech & Tobin 1981). To our knowledge, no previous serologic studies of CMV infection have been reported from Bolivia. The present study was undertaken to determine the prevalence of CMV infection in the Santa Cruz region, south-eastern Bolivia. PMID- 2558226 TI - Poliomyelitis: 20 years--the Pondicherry experience. AB - During a 20-year period, 12,830 cases of acute poliomyelitis in children (aged 1 month-12 years) were seen in a South Indian teaching hospital (Pondicherry). Almost three-quarters (72.4%) of children were below the age of 2 years. The median age at onset of poliomyelitis shifted from 19.5 months (in the period 1968 1977) to 11 months (in the years 1978-1987). After 1981, epidemic peaks were noted every third year. 66.3% of cases had not received even a single dose of oral polio vaccine and 14.7% cases were vaccine failures (in the years 1983 1987). Provocation of poliomyelitis occurred in 66% of children and usually followed intragluteal injections associated with treatment of non-specific fevers. The overall mortality was 2.13%, the major cause of death being respiratory failure (60%). Polio virus type 1 was the major isolate among selected cases studied during 1983-1987. The risk factors identified in our study are lack of immunization and administration of intramuscular injections during the pre-paralytic phase. Ensuring early administration of multiple doses of oral polio vaccine to infants and avoiding intramuscular injections in young febrile children are important measures for the control of poliomyelitis--a continuing problem in Southern India. PMID- 2558227 TI - Hepatitis A antibodies: prevalence in Saudi Arabia. AB - Hepatitis caused by hepatitis A virus (HAV) occurs at a variable prevalence in different countries, and the exposure to HAV is as high as 100% in some areas. There appears to be a close association between the prevalence of anti-HAV and socioeconomic status and age. In this paper, we report the prevalence of antibody to HAV (anti-HAV) in five regions covering different parts of Saudi Arabia. The results of this study show that the overall prevalence of anti-HAV is 96% and 95.1% in the total male and female population respectively. In different age groups the prevalence varies from 78-100% with a concomitant increase with age. These results show that the rate of exposure to HAV is high in Saudi Arabia and predominant exposure occurs in early childhood. PMID- 2558229 TI - [A case of adenoid cystic carcinoma of the esophagus]. AB - A surgical case of an adenoid cystic carcinoma (ACC) of the esophagus in a 75 year-old man is reported. Histologically, the tumor consisted of an ACC, a squamous cell carcinoma and a small tubular adenocarcinoma. The ACC and the tubular adenocarcinomatous regions in the submucosa and the lamina propria were continuous with the overlying squamous cell carcinoma and atypical squamous epithelium. Immunohistochemically, two types of tumor cells were detected in the ACC. One was found to have EMA-positive epithelial cells whereas the other had actin-positive myoepithelial cells. In the normal esophageal gland, actin positive cells are found at the periphery of acini and around the layer of epithelial cells in the small duct but they have not been detected in the main duct. These findings suggest that the tumor developed from the small duct and differentiated into two directions: an ACC and a squamous cell carcinoma. PMID- 2558228 TI - [A clinico-pathological study of hepatocellular carcinoma patients with other primary malignancies]. AB - In examining 293 patients with a hepatocellular carcinoma (HCC), 27 patients (9.2%) were found to have another primary malignancy located elsewhere. Of this number, fifteen were male and twelve were female, with ages that ranged from 53 to 81 years (mean 67.3 + 8.9 years). Twenty-four of these patients were found to have two other malignancies, where in 13 had both malignancies with a one year interval and other 11 after more than one year. The remains, three patients, had triple malignancies. These associated cancers included eight gastric cancers, four uterine, four colon cancers and other such cancers. The proportion of females with double cancers was higher than in those with only an HCC, although there were no difference in the HBsAg, age, past history of blood transfusion, the positive rate of familial malignancy, and the HCC between those with solely an HCC and those with other cancers as well. This suggests that the occurrence of an HCC concomitant with or followed by the development of another primary malignancy is not rare. PMID- 2558231 TI - [Pseudomyxoma peritonei with myxoglobulosis--a case report and the pathogenesis of the myxoglobules]. AB - A case of pseudomyxoma peritonei with myxoglobulosis is described. The myxoglobulosis was characterized by globules consisting of firm opaque whitish gray nodule that measured up to 1.0 cm in diameter. Many globules in the abdominal cavity were filled with mucinous material. Electron microscopically, these globules were found to be covered by mesothelial cells with microvilli and contained mucous material, fibrous tissue, and a few myofibroblasts. PMID- 2558230 TI - [A case of hepatocellular carcinoma (HCC) with bleeding due to duodenal perforation by the tumor]. AB - On June 11, 1986, a 70-year-old man was introduced to our hospital because of an elevated AFP and hepatomegaly. He was diagnosed as having an HCC in the left medial segment and a transcatheter arterial embolization (TAE) was able to reduce his AFP level. In December, 1986, repeated tarry stool was noted, and he was readmitted to hospital on January, 28, 1987, because of severe anemia. An ordinary X-ray revealed an abnormal gas shadow in the right upper abdomen. A subsequent endoscopic examination showed a tumoral mass protruding into the duodenal lumen through a duodenal perforation. After death an autopsy revealed that the perforation was due to the expansive growth of the tumoral mass to the duodenum. PMID- 2558233 TI - [Malignant lymphoma displaying rearrangements of both immunoglobulin and T-cell receptor genes]. AB - A 65-year-old male was admitted to our hospital due to recurred malignant lymphoma of left tonsil origin. Studies using flow cytometry on mononuclear cells in peripheral blood revealed the appearance of intermediate B cells, and examinations on gastrointestinal tract showed diffuse infiltration of medium sized lymphoid cells into stomach and colon, that had the same phenotype as tumor cells in peripheral blood. They suggested leukemic change and gastrointestinal tract infiltration of malignant lymphoma. Southern blot analysis revealed T-cell receptor beta-chain gene rearrangement as well as immunoglobulin gene rearrangement. Analysis by histo in situ hybridization on infiltrated gastric mucosa specimen showed diffuse expression of immunoglobulin mRNA in tumor cells, but no message of T-cell receptor beta-chain gene. Northern blot analysis showed the same result. It suggests that in this case, the rearrangement of T-cell receptor beta-chain gene is ineffective rearrangement without transcription to mRNA. PMID- 2558232 TI - [Interstitial pneumonia (IP) in bone marrow transplantation in leukemia--120 cases analysis in Nagoya Bone Marrow Transplantation Group]. AB - Results of the bone marrow transplantation (BMT) for 120 cases of leukemia, which were done in nine institutes in Nagoya (Nagoya Bone Marrow Transplantation Group) last ten years, were analyzed to determine the factors associated with an increased risk of developing interstitial pneumonia (IP). IP developed 49 out of 120 patients (49.8%) and case fatality rate was 63.3%. The median time from transplantation to onset of IP was 81 days (range 13-575 days), in 30 out of 49 cases (61.2%), this complication developed within 100 days after transplantation. Of the 49 patients who developed IP, cytomegalovirus (CMV) infection was associated in 18 cases (36.7%), no cases of P. carinii infection was detected. Five factors were associated with an increased risk for developing IP, (1) older age (greater than or equal to 47.0%: less than 10 y. 10.0%) (p less than 0.01) (2) disease stage at BMT (non-remission 76.2%: remission 32.5%) (p less than 0.01) (3) presence of acute GVHD ((+) 52.5% (-) 28.8%) (p less than 0.05) (4) onset day after BMT (less than or equal to 100 days 61.2%: greater than 100 d. 38.8%) (p less than 0.01) (5) sex matching between donor and patient (sex match 28.8%: sex mismatch 57.1%) (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558234 TI - [Philadelphia-positive acute mixed leukemia with monosomy 7]. AB - A 26-year-old male was admitted to our hospital because of fever and leukocytosis. On admission, a white blood cell count was 28,300/microliters with 46.5% blast cells and 16.0% atypical monocytoid cells, a hemoglobin level 13.7 g/dl, and a platelet count 15.0 X 10(4)/microliters. Bone marrow contained 58.8% of peroxidase-negative blast cells. He was diagnosed as acute lymphoblastic leukemia (ALL L2) according to the FAB classification. Chromosome analysis revealed the marrow cells to contain 45, XY, -7, t(9; 22) (q34; q11). On surface marker analysis, the leukemic cells were positive for both lymphoid (CD10) and myeloid markers (CD13). Two color flow-cytometric analysis showed two distinct populations with CD10 and CD1 3, respectively. Rearrangements of both immunoglobulin heavy chain and T cell receptor beta-chain were observed. The "breakpoint cluster region" on chromosome 22 was not rearranged. On the basis of these findings, we thought this case being acute mixed leukemia. He was refractory to AdVP therapy and BHAC-DMP therapy. He is now under treatment with A Triple-V therapy. PMID- 2558235 TI - [Specific cytoplasmic crystalline inclusions in multiple myeloma with amyloidosis]. AB - It is widely known that multiple myeloma is sometimes followed by amyloidosis. It is also not particularly rare that inclusions exist in myeloma cells. However, there has been no previous report of a case of myeloma with both inclusions and amyloidosis. A 60-year-old female initially complained of a tendency to bleed, which was caused by fibrinolysis. Amyloid deposition in bone marrow stroma and the gastric submucosa was recognized, in addition to crystalline inclusion in the cytoplasm of myeloma cells. An immunoelectron microscopic study demonstrated the amyloid fibrils and the crystals to react positively to anti-lambda serum. No crystals were found in macrophages, and no relationship was recognized between lysosomes and crystalline inclusions in the cytoplasm of myeloma cells. This case had disturbed transportation or secretion of the lambda type L chain and it was considered that crystals derived from the lambda type L chain were formed in the cytoplasm of myeloma cells. PMID- 2558236 TI - [Childhood Ki-1 lymphoma complicated with multiple bone destruction]. AB - A 4-year-old girl was admitted because of fever, swelling of left chest wall and left axillary lymphadenopathy. Chest XP revealed left pleural effusion. Ga scintigram showed multiple accumulation in skull, left ribs and iliac bone. A diagnosis of childhood Ki-1 lymphoma was made from the pathological findings of tumor in the skull. Immunopathological study revealed that the neoplastic cells were CD8 positive (suppressor phenotype). PMID- 2558239 TI - [Molecular biology of 1,25(OH)2D3 receptor and vitamin D response element]. PMID- 2558237 TI - [Thyrotropin (TSH)--gene organization and expression]. PMID- 2558238 TI - [Beta-adrenergic receptor structure]. PMID- 2558240 TI - [Turnover of inositol phospholipids and intracellular messenger systems]. PMID- 2558241 TI - Herpes simplex virus in oral mucosal ulcers in patients with hematological malignancy. AB - Developments of oral mucosal ulcers induced by herpes simplex virus (HSV) were studied in patients with hematologic malignancy. Herpes simplex virus type-1 (HSV 1) was identified by immunological staining using virus-specific monoclonal antibodies in the epithelial cells of such ulcers from two patients with malignant lymphoma (ML), three with acute myeloblastic leukemia (AML), one with refractomy anemia with excess blasts, two with chronic myelocytic leukemia (CML), one with acute lymphoblastic leukemia (ALL) and one with aplastic anemia (AA). Herpes simplex virus type-2 (HSV-2) was also identified in an ulcer from a patient with AML. Isolation of HSV-1 was successful in the two patients with ML, one with CML, one with AML, the one with ALL and the one with AA. The ulcers developed on the tongue (four cases), buccal membrane (five cases), hard palate (one case), soft palate (one case), soft palate (one case) and gingiva (two cases). Only one patient with CML and one with AML had accompanying labial vesicular lesions. All patients except the one with AA had previously been given combination chemotherapy with anti-neoplastic agents. The results indicate that HSV may have an important role to play in the development of chemotherapy-related oral mucosal ulcers in patients with hematological malignancy. PMID- 2558242 TI - Parathyroid hormone-related protein and transforming growth factor activities in an extract from a breast cancer associated with humoral hypercalcemia of malignancy. AB - The pathogeneses of hypercalcemia and hypophosphatemia which developed in a patient with metastatic invasive ductal breast carcinoma were studied. The patient had low plasma levels of immunoreactive parathyroid hormone (PTH) and 1,25(OH)2D, increased nephrogenous cyclic adenosine monophosphate (cAMP) excretion and low TmPO4/GFR, suggesting the presence of humoral PTH-like activity. The tumor extract showed activities which would stimulate bone resorption in vitro and cAMP generation in the osteogenic cell line, MC3T3 E1, and in the rat kidney cortex. In addition, the extract stimulated epidermal growth factor (EGF)-independent colony formation of the NRK 49F cells in soft agar, and inhibited the binding of EGF to A431 cells, indicating it to have transforming growth factor (TGF)-alpha activity. The extract contained appreciable amounts of immunoreactive PTH-related protein (PTH-rP) but negligible amounts of immunoreactive PTH. Thus, the PTH-like activity for stimulating cAMP generation in the bone and kidney was attributed to PTH-rP. Chromatographic analyses on reverse phase high performance liquid chromatography (HPLC) separated the PTH-rP activity from that of TGF-alpha and the bone resorbing activity in vitro was found only in the fractions of PTH-rP. It was concluded that this breast cancer produced PTH-rP as well as TGF-alpha, and the former was thought to have a major role to play in the humoral hypercalcemia of malignancy observed in this patient. PMID- 2558243 TI - [Clinical evaluation of serum CA-50 and CA 19-9 in gynecological tumors]. AB - We conducted a study to evaluate cancer associated antigen CA-50 and CA 19-9 as tumor markers of gynecological malignancies. The positive rates of CA-50 and CA 19-9 for ovarian cancer were 35.5% and 48.8%, respectively, and thus were not very high. In terms of histological typing, relatively high positive rates were noted in mucinous type-42.9% for CA-50 and 71.4% for CA 19-9. Both antigens showed high false positive rates for benign ovarian tumors, especially for dermoid cyst, but produced few false positive cases of endometrial cyst. For cervical cancer and endometrial cancer, these antigens were positive at low rate. In conclusion, the present evaluation indicated that both CA-50 and CA 19-9 do not necessarily suffice as screening markers for gynecological malignancies; that they could potentially be of help for diagnosis of ovarian cancer of mucinous type; and that their false positive rates for endometrial cyst were very low. PMID- 2558244 TI - Flow cytometric analysis of the nuclear DNA content of hepatocellular carcinoma. AB - The nuclear DNA content of 77 resected specimens from 65 cases of hepatocellular carcinoma (HCC) was measured by means of flow cytometry. The DNA index (DI) was calculated and the correlation between the DNA ploidy pattern and clinicopathological findings was studied. In the cases of HCC with a diameter of less than 5 cm, the 3-year survival rate of the aneuploid cases was 44.5 per cent, which was significantly lower than the 91.4 per cent of the diploid cases (p less than 0.001). Serum AFP levels were over 1000 ng/ml in 46.4 per cent of the aneuploid tumors and 18.5 per cent of the diploid tumors (p less than 0.05). The DI's were investigated in several sites of the same tumor and no difference was seen among the different sites in 16 out of 17 tumors. From 8 recurrent cases out of 12 who underwent a second resection, seven did not show any significant differences in DI from their primary tumor. On the other hand, four cases of second primary tumors showed different DI's to those of their first primary tumor. Intra-hepatic metastatic tumors exhibited the same DI's as their primary tumors. Thus, the nuclear DNA ploidy pattern may serve as a stable and valuable marker in predicting the malignant potential and prognosis of HCC. PMID- 2558245 TI - Multimodality therapy for small cell carcinoma of the lung--the role of surgical treatment. AB - Reviewing the outcome of 70 cases of clinically localized small cell lung cancer (SCLC) treated with combined modality treatment, we attempted to define the role of resection in this disease. The survival rate for all cases was 37 per cent at 2 years and 23 per cent at 3 years with a median survival time (MST) of 14 months. For 25 resected cases the overall 5-year survival rate was 37 per cent with an MST of 26 months. According to clinical staging, 5-year survival was 64 per cent for stage I and 20 per cent for stage II. However, none of the stage III cases achieved long-term survival, of over 3 years. In 45 non-resected cases, the overall response rate was 84 per cent with a 44 per cent complete response. The overall survival rate was 27 per cent at 2 years and 14 per cent at 3 years with an MST of 11 months. The 20 cases who achieved complete response had an MST of 26 months with 51 per cent alive at 2 years and 19 per cent at 5 years. Thus, we consider that lung resection is definitely indicated in cases with stage I and probably stage II SCLC. For stage III, however, particularly in cases with N2 disease, resection seems to offer no special benefit in favor of survival compared to combination chemotherapy and radiotherapy. PMID- 2558246 TI - Double cancer of the liver and stomach with situs inversus totalis--a case report. AB - A case of a 66-year-old woman with situs inversus totalis who developed hepatocellular carcinoma (HCC) as well as stomach cancer, is reported herein. The patient was successfully treated with a left hepatic lobectomy for the HCC and a B-I gastrectomy for the stomach cancer. Careful anatomical mapping made it possible to perform a combined resection of the liver and stomach in the presence of this congenital anomaly. PMID- 2558247 TI - [Tracheobronchial reconstruction of malignant tumors]. AB - To August, 1988, we performed tracheobronchial reconstruction for 159 patients with malignant tumors. Operative methods in these patients and the operative results studied in the end of March, 1989, was reported. As for adenoid cystic carcinoma, we stressed, by illustrating cases, that postoperative radiation therapy was useful and that it should be given for all patients with this disease. About one half of patients with thyroid carcinoma infiltrating the trachea died of symptoms related with tracheal stenosis. However, tracheobronchial reconstruction improved 10 year survival rate in patients with advanced thyroid carcinoma remarkably, which became close to that in patients with non-infiltrating thyroid carcinoma. As for lung cancer, tracheobronchial reconstruction had good indication for squamous cell carcinoma. PMID- 2558248 TI - Enhancement of neutralizing activity of anti-feline herpesvirus type 1 sera by complement supplementation. PMID- 2558250 TI - Pathogenicity of a reovirus type 2 in suckling mice: insulitis and lymphoid atrophy. PMID- 2558249 TI - Enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to glycoprotein antigen of bovine herpesvirus type 1. PMID- 2558251 TI - Characteristics of in vitro passaged cells derived from a rat transplantable malignant fibrous histiocytoma. AB - Morphologic and functional characteristics were investigated on in vitro passaged cells (MT-P) derived from a rat transplantable malignant fibrous histiocytoma (MFH-MT). There were spindle, polygonal, and giant cell types in MT-P. Ultrastructurally, the polygonal and giant cells had the abundant cytoplasm with many lysosomes and processes, whereas the spindle cells possessed smooth cell surface and a small number of lysosomes in their cytoplasm. Immunorosette formation for Fc- and C3-surface receptors and phagocytic activity were demonstrated in 10-20% of MT-P. MT-P were positive for acid phosphatase, nonspecific esterase and alkaline phosphatase. Chromosomes counted in 100 MT-P ranged from 32 to 100 with two peaks of 64 and 76. Tumors induced in syngeneic rats by inoculating MT-P showed variable histologic patterns. They were composed partly of histiocytic cells arranged in a compact sheet. Fibroblastic cells often arranged in a storiform pattern or were supported by myxoid matrix. Osteosarcoma like structures were occasionally found in the tumors. These results suggest that MFH-MT is heterogeneous, although some cells constituting the tumors have histiocytic markers. PMID- 2558252 TI - Immunocytochemical and morphometric studies on the pars distalis of the golden hamster from perinatal to senile stage. AB - Six cell types of the pars distalis were studied from perinatal to senile stage in the golden hamster in combination with morphometric analysis and immunocytochemistry. GH cells occupied only a small area at 15 days of gestation and increased remarkably after birth to occupy a significantly larger area in the young adult than that in the other stages. PRL cells first appeared at 3 days after birth and increased rapidly thereafter to be distributed throughout the pars distalis. They always occupied a larger area in females than in males from 3 weeks after birth onward. ACTH cells were distributed mainly in the peripheral region of the pars distalis. They were dominant at 15 days of gestation, but relatively decreased after birth. The gonadotrophic cells were divided into LH cells and FSH cells. About 70% of LH cells were also immunoreactive to anti-FSH serum. LH cells occupied a larger area than that of FSH cells in every stage examined. TSH cells occupied the second largest area at 15 days of gestation, but rapidly decreased at 3 weeks after birth to occupy the smallest area among the immunoreactive cell types throughout the life-span. The chronological changes in the percentages of all cell types constituting the pars distalis seemed to reflect the activity of these cell types at a given stage. PMID- 2558253 TI - Adrenocorticotropin and cortisol levels in the plasma of bovine fetuses and neonates. AB - Plasma ACTH and cortisol levels in the bovine fetuses over the period of 5 to 9 months of gestation and in the neonates immediately after birth and at 5 days old were studied. In the bovine fetuses, the plasma ACTH levels ranged from 60.8 +/- 17.8 to 71.3 +/- 19.7 pg/ml over the period of 5 to 7 months of gestation. It increased rapidly to 239.2 +/- 261.5 pg/ml at 8 months and to 406.9 +/- 409.4 pg/ml at 9 months of gestation. In the neonates immediately after birth it decreased to 182.3 +/- 110.7 pg/ml. The plasma cortisol levels ranged from 3.23 +/- 2.12 to 3.85 +/- 2.52 ng/ml over the period of 5 to 8 months of gestation and increased to 8.10 +/- 4.88 ng/ml at 9 months of gestation. It then increased rapidly to 88.35 +/- 42.78 ng/ml in the neonates immediately after birth. The correlation between plasma ACTH and cortisol levels in the fetuses of 5 to 7 months of gestation was not significant, but in the fetuses of 8 and 9 months of gestation and neonates were significant. However, especially immediately after birth, the increase in plasma cortisol occurred without a concomitant rise in plasma ACTH. According to these findings, it is suggested that the pituitary adrenocortical axis in bovine fetus matures in the later stage of gestation and an increase of sensitivity in the fetal adrenal gland to ACTH may serve as a trigger for the onset of parturition. PMID- 2558256 TI - A SepPak HPLC method for tricyclic antidepressant drugs in human vitreous humor. AB - Death from tricyclic antidepressant overdose has become an all-too-common occurrence. Several factors, including postmortem concentration changes, can render blood and tissue samples useless for the determination of antidepressant drug concentrations. We present here an efficient method of solid-phase extraction for these drugs from vitreous humor and a reversed-phase, isocratic high-performance liquid chromatographic method for the simultaneous quantitation of amitriptyline, doxepin, and imipramine and their desmethylated metabolites. PMID- 2558257 TI - Full length L1 retroposons contain tRNA-like sequences near the 5' termini- hypothesis on the replication mechanism of retroposons. AB - Retrotransposons replicate via a complex mechanism which depends on, among other things, the presence of long terminal repeats (LTRs) and a tRNA binding site just 3' of the 5' LTR. The LINES 1 (L1) family of sequences, which similar to retrotransposons in many other properties, represents a new class of retroposon which does not possess LTRs. However, we show here that the repetitive 5' motif associated with murine L1 elements contains a tRNA-like sequence in a location analogous to the position of the retro-transposon tRNA binding site. Although the repetition of such a 5' motif has only been found associated with murine L1 elements, we have found an analogous tRNA-like sequence near the 5' ends of the L1 elements from each of the other analyzed species for which the L1 family has been characterized, that is rat (L1Rr), human (L1Hs), drosophila (I element) and trypanosome (INGI). The conservation of this tRNA-like sequence near the 5' terminus of L1 elements from such diverse species suggests that it plays a functional role in the life of the L1 class of retroposon. PMID- 2558254 TI - [Sequential induction chemotherapy and radiation treatment of inoperable small cell bronchial cancer. Results of a prospective randomized study]. AB - To study the potential benefit of sequential chemotherapy in inoperable small cell lung cancer (SCLC), from 1982 to 1986 ninety-one patients with histologically proven and previously untreated SCLC (median age: 53 years; median Karnofsky status: 80%) were randomly assigned to an initial therapy with adriamycin (since 1984 epirubicin), cyclophosphamide, vincristine (ACO resp. EPICO) or etoposide/cisplatin (VP16/DDP). Treatment courses were repeated every 3 weeks for a total of less than or equal to 6 courses with a crossover after a maximum of 3 cycles of either regimen. Limited disease (LD) patients with bronchoscopical, computertomographical and (re-) mediastinoscopical complete remission (CR) randomly received either a thoracic irradiation with 40 Gy or observation only. Overall, 60 out of 85 evaluable patients achieved an objective remission. A CR was observed in 24/51 patients (47%) with limited disease, and in 8/34 patients (24%) with extensive disease. Both, ACO (EPICO) and VP16/DDP were equally effective as initial and second-line therapy. Moreover, after failure to the initial therapy an objective remission could be achieved in 13% of the patients following the alternative second line combination. In 28% of LD patients with an otherwise complete remission residual tumor was detected by (re-) mediastinoscopy. Median survival times were 14 (CR: 16) months in LD patients and 10 (CR: 15) months in ED patients. At present, median survival is significantly improved in irradiated versus non-irradiated LD patients (25 vs. 13 months, p less than 0.04). The remission rates and median survival times observed in this study are comparable to those of a historical control group treated with ACO plus radiotherapy alone. PMID- 2558255 TI - [Relation between post-thrombotic syndrome, ADP-induced thrombocyte aggregation and intrathrombocyte calcium content]. AB - Correlation between postthrombotic syndrome, ADP-induced aggregation and intracellular calcium concentration. This study presents a comparison of the ADP induced platelet aggregation and free cytosolic platelet calcium concentration between patients with postthrombotic syndrome and healthy volunteers. The half maximal effective dose of the platelet aggregation induced by ADP was significantly decreased in postthrombotic syndrome (p less than 0.005). The mean values were 0.42 mumol/l in postthrombotic syndrome (n = 22) and 1.00 mumol/l in normal controls (n = 25). The free intracellular calcium concentration was slightly but significantly diminished in patients compared with healthy volunteers. In both groups there was seen a remarkable interindividual spread of platelet calcium concentrations. There was seen no significant correlation between cytosolic calcium concentration of platelets and the half maximal effective dose of ADP-induction. Therefore the considerable enhanced aggregability in postthrombotic syndrome cannot explained with altered cytosolic calcium content of platelets. PMID- 2558258 TI - A mathematical modelling study of epiphyseal cartilage calcification. AB - Recent studies in this laboratory have suggested that proteoglycan may function as a Ca ion-exchanger in the calcification of epiphyseal growth plate cartilage. Specifically, it has been proposed that phosphate liberated from hypertrophic chondrocytes may displace calcium ions bound to the anionic groups of proteoglycans, thereby raising the Ca x PO4 activity product above the threshold for precipitation of hydroxyapatite. In order to determine whether this mechanism is quantitatively feasible, a mathematical model of the interaction between Ca, Na, proteoglycan and phosphate has now been developed. This model is based on a general binding theory, and utilizes previously-determined values for the binding constants of the Ca-proteoglycan interaction, inhibition constants for the effect of Na and phosphate on this interaction, and literature values for the concentrations of proteoglycan, Na and Ca in epiphyseal cartilage. Using this approach, it was predicted that the free Ca concentration in epiphyseal cartilage in the absence of phosphate will be 1.55 mM. At 0.7 mM phosphate, the approximate concentration in non-calcified cartilage matrix, the free Ca concentration will be 2.40 mM, corresponding to a Ca x PO4 product of 1.68 (mM)2. In order to achieve a Ca x PO4 product sufficient for spontaneous precipitation of hydroxyapatite [approximately 4.3 (mM)2], a phosphate concentration of approximately 1.40 mM is required. Therefore, calcification of epiphyseal cartilage matrix by the mechanism described above will require an approximate doubling of the phosphate concentration in the pre-calcifying zones, indicating that the release of a fraction of the intracellular phosphate could trigger the calcification process. PMID- 2558259 TI - Ph-negative T cells in a patient with chronic myelogenous leukemia for twenty eight years. AB - The frequency of metaphases without a Philadelphia chromosome was determined in mitogen-stimulated cultures of peripheral blood mononuclear cells (PBMC) and purified T lymphocytes (93% CD2-positive) from a patient with chronic myelogenous leukemia (CML) for 28 years. The PBMC cultures contained few Ph-negative cells (8%), but they constituted 92% of the metaphases in T cell cultures, indicating few if any Ph-positive T cells in the patient's circulation. The results demonstrate that T cells derived from the leukemic clone may fail to replace the non-neoplastic population even when CML arises in childhood and the patient survives for many years. This raises questions concerning the normal role of the bone marrow as a source of T cells after infancy, and also whether Ph-positive lymphocytes may be at a disadvantage for growth. PMID- 2558260 TI - Double labeling of cytochrome oxidase and gamma-aminobutyric acid in central nervous system neurons of adult cats. AB - The relationship between the levels of cytochrome oxidase and gamma-aminobutyric acid (GABA) was investigated within single neurons by double labeling the 2 markers in the same section. Double staining was equally effective when immunogold-silver staining of GABA was followed by indirect immunoperoxidase labeling of cytochrome oxidase, or when cytochrome oxidase histochemistry was followed by immunogold-silver staining of GABA. Neurons in the perigeniculate nucleus (PGN) and basket cell terminals in the cerebellum were GABA positive and rich in cytochrome oxidase. Interneurons of the lateral geniculate nucleus (LGN) as well as stellate and Golgi cells of the cerebellum were GABA-rich but poor in cytochrome oxidase. These results demonstrate that there is no consistent relationship between the levels of cytochrome oxidase and GABA in neurons. PMID- 2558261 TI - [Detection of early cytomegalovirus antigen in cell culture]. AB - The reference technique for the diagnosis of active cytomegalovirus infection is the isolation in cellular culture. Its major drawback is the interval between the inoculation of the sample and the development of the characteristic cytopathic effect. Occasionally, this delay may be longer than four weeks. The centrifugation of the sample on the cell monolayer at the time of inoculation and the use of a fluorescein-labeled monoclonal antibody for the detection of the early antigen in cells may considerable reduce the time required for the diagnosis of cytomegalovirus infection. In the present study the technique of detection of the early antigen by immunofluorescence was compared with conventional cell culture in 258 clinical samples referred to the laboratory for study. Fifty-one of them were positive: 28 with both techniques, 12 only with cell culture and 11 only with immunofluorescence. The mean time to obtain positive results was 25 hours for immunofluorescence and 13 days for culture. PMID- 2558262 TI - [Hemolytic anemia caused by pyrimidine 5'-nucleotidase (P5N) deficiency 15 years later. Apropos of 2 new cases of hereditary deficit and another one of lead poisoning]. AB - Congenital pyrimidin 5'nucleotidase deficiency manifests as hemolytic anemia with basophilic stippling. In lead poisoning, anemia, basophilic stippling and inhibition of erythrocyte pyrimidin 5' deficiency are also observed. In the present work, we report two cases of hemolytic anemia secondary to congenital deficiency of pyrimidin 5' nucleotidase and another case secondary to lead poisoning. Since 1974, when pyrimidin 5' nucleotidase deficiency was isolated, is known that hemolysis is related to the accumulation of pyrimidin nucleotides within the erythrocytes that behave as metabolic inhibitors. However, the precise metabolic process whose inhibition leads to the shortening of erythrocytes half life has not been elucidated yet. PMID- 2558264 TI - Sensory recovery in the plantar aspect of the foot after surgical decompression of posterior tibial nerve. Possible role of steroids along with decompression. AB - In leprosy, involvement of the posterior tibial nerve leads to sensory loss in the plantar aspect of the foot. As a result plantar ulcers are common and lead to deformity and disability. Restoration of plantar sensation can prevent ulcer formation. Posterior tibial decompression was done for the recovery of sensation in the plantar aspect of the foot. Seventy-two patients under went decompression on 84 feet, 25 received steroids pre- and post-operatively. The recovery of sensation was better if surgery was done before 6 months of onset of anaesthesia. Decompression along with steroids gave better results than decompression alone in patients with active neuritis especially in BT cases whereas in BB, BL and LL cases there was no significant improvement of sensation. The results are discussed. PMID- 2558263 TI - [Dementia caused by miliary cerebral metastasis of a hepatocarcinoma]. AB - Different types of tumors account for the etiology of 8.6% of all cases of dementia. Cerebral computed tomography (CT) permits the detection of most of them. The miliary metastasis variety commonly presents as dementia, and CT may then be normal. A patient with late epilepsy after cerebral infarction developed subacute dementia and he subsequently died. Post mortem study disclosed miliary cerebral metastases of a clinically unsuspected hepatocarcinoma. Cerebral CT did not detect the metastases. Although the incidence of hepatocarcinoma is increasing, we are unaware of any similar reported case, as cerebral involvement is exceptional in this condition. Despite its rarity, this case highlights the need to perform thorough neuropathological investigations in dementia. PMID- 2558266 TI - Insulin-like growth factor (IGF)-I and IGF-binding activity in normal and fast growing chickens. AB - Insulin-like growth factor(IGF)-binding activity were characterized in sera normal, fast growing, and very fast growing chicken strains. In contrast to a previous report, specific IGF-binding activity was observed in this nonmammalian species. Age-related IGF-binding activity levels did not differ between strains. IGF-I levels were significantly higher for the normal as compared to the 2 fast growing strains. Chromatographic studies show that IGF-I is associated with acid dissociable high MW complexes in chicken serum, and the MW patterns are similar to human serum. Circulating levels of IGF-I or IGF-binding activity do not account for accelerated somatic growth in inbred large-bodied chicken strains. PMID- 2558265 TI - Kappa opioid receptors of human placental villi modulate acetylcholine release. AB - Human placental villus tissue is non-innervated, yet it contains components of the opiate and cholinergic systems. We investigated whether opioids modulate a calcium dependent acetylcholine release from the villus tissue in a manner similar to that demonstrated by the parasympathetic nerve-smooth muscle junction. We reported that the kappa receptor agonist ethylketocyclazocine (EKC) inhibits acetylcholine release, and that the inhibition is reversed by the selective antagonist, Mr2266. Findings reported here substantiate the role of opioids as modulators of acetylcholine release from villus tissue. The nonselective agonist, morphine, also inhibits acetylcholine release. Inhibition caused by morphine is reversed by low concentrations of non-selective antagonists, naloxone and naltrexone. Naloxone at high concentrations potentiates the inhibition of acetylcholine release caused by morphine. In addition, the calcium channel blocker, diltiazem, was found to inhibit the release of acetylcholine. The combination of morphine and diltiazem resulted in a greater inhibition of acetylcholine release than by either alone. These results suggest that opiate cholinergic interactions occur in non-neural tissue with a mechanism similar to that known to occur at certain cholinergic synapses. PMID- 2558267 TI - The effect of mycoplasma on the autocrine stimulation of human small cell lung cancer in vitro by bombesin and beta-endorphin. AB - The tumor stem cell clonogenic assay was utilized to investigate the autocrine growth response of small cell lung cancer (SCLC) to bombesin (BN) and beta endorphin (beta-E). Mycoplasma contamination was detected in the human SCLC cell line NCl-H345 by a nucleic acid hybridization assay which detects mycoplasma ribosomal RNA. Clonogenic assays of mycoplasma (+) cells were compared to assays of the same cell line following treatment for mycoplasma. Concentrations of beta E ranging from 0.1nM to 25nM or BN (0.1nM-100nM) were added to cells, media and agarose and applied to prepared base layers. Following incubation for 12-14 days at 37 degrees C, the degree of clonal growth stimulation was determined by colony counts greater than or equal to 42 mu. The non-infected cell population grew in the presence of 25nM BN up to 69% over control growth. The infected cells, however, did not grow more than 27% above control. In the presence of 10nM beta E, colony counts of non-infected cells exceeded the control values by up to 187% whereas the mycoplasma (+) colonies did not grow more than 20% over the control values. These results indicate a marked reduction in the response of SCLC cell lines to the peptides BN and beta-E when infected with mycoplasma. Since infecting mycoplasma typically adhere to cellular membranes, these adherent mycoplasma may interfere with membrane receptors or alter signal transduction, thus, inhibiting the development of the autocrine response. PMID- 2558269 TI - [Flumazenil (a specific antagonist of benzodiazepine receptor]. PMID- 2558268 TI - Sanded nuclei in delta patients. AB - Sanded nuclei are nuclei with eosinophilic inclusions identified by light microscopy in cases of chronic hepatitis B virus infection. In hematoxylin and eosin-stained sections, these inclusions have an almost homogeneous, finely granular texture giving a sandy appearance. They have been related to excess hepatitis B core antigen formation. We have studied liver biopsies from two HBsAg positive immunosuppressed patients with numerous sanded nuclei, morphologically identical to those previously described in hepatitis B. Immunohistochemically, sanded nuclei showed a strong nuclear positivity for delta antigen, but were negative for hepatitis B core antigen. Hepatitis B core particles were not demonstrable by electron microscopy. To our knowledge, this is the first time that sanded nuclei have been related to hepatitis delta virus (HDV) infection. PMID- 2558270 TI - Use of host modified bacteriophages in development of a phage typing scheme for Clostridium perfringens. AB - A bacteriophage isolated from a faecal strain of Clostridium perfringens was adapted to a number of host strains of clinical swab and faecal isolates. A typing scheme was developed using nine host modified phages. Of 109 strains the phage types of 57 (52.3%) were identified. Nine (8.2%) other strains were sensitive to the phages at varying degrees. The remaining 43 (39.4%) strains were resistant. Eleven of the 57 typable strains yielded cell-surface mutants which belonged to different phage types from their parent strains. The phages were applied at two different dilutions in a reverse spotting method to eliminate false positive reactions due to inhibition by phage adsorption or bacteriocins. The method was simplified with the aid of a multipoint inoculator for routine use. PMID- 2558271 TI - Screening for cervical carcinoma. AB - At present, cytological examination of cervical smears is the method used to screen the female population for cervical abnormalities. This employs the Papanicolaou staining technique and cell morphology to detect abnormal cells from the susceptible transformation zone of the cervix. However, with the recent discovery of a possible link between human papillomavirus (HPV) infection and cervical cancer, a test to determine the presence of HPV infection may be appropriate. Tests are still at a developmental stage and at present used only for research purposes. Various DNA-DNA or DNA-RNA hybridisation techniques are employed in experimental screening studies, and these involve probing cells or DNA extracted from cells for HPV DNA using radio-labelled probes. PMID- 2558272 TI - Phosphodiesters in the liver: the effect of field strength on the 31P signal. AB - 31P NMR spectra of the rat liver were recorded in vivo at 2.35 and 8.5 T. There was a large peak in the phosphodiester region of spectra obtained at 2.35 T which was much reduced at 8.5 T. The peak at 2.35 T is unlikely to be primarily from free cytosolic phosphodiesters, which would not be expected to display such a marked field dependence. PMID- 2558273 TI - Proton magnetic resonance of paramagnetic metalloproteins. PMID- 2558274 TI - Determination of equilibrium constants of enzyme-bound reactants and products by nuclear magnetic resonance. PMID- 2558275 TI - Spin labeling of proteins. PMID- 2558276 TI - Quantitative evaluation of receptor-mediated endocytosis. PMID- 2558278 TI - Susceptibility of laboratory and domestic animals to experimental infection with Potiskum virus. AB - The biological characteristics of Potiskum virus, a hitherto undescribed virus isolated in Nigeria from the liver of a giant rat (Cricetomys gambianus), were studied by experimental infections of laboratory and domestic animals. The laboratory animal hosts used included mice, rats, rabbits and chicks. Suckling and weaning mice succumbed to fatal infection when infected with Potiskum virus by intracerebral or intraperitoneal routes. Infected mice had high titres of virus and mild histopathological lesions which were confined to the brain. Chicks also developed a fatal disease following subcutaneous or oral infections with Potiskum virus. In contrast, albino rats and rabbits failed to succumb to overt disease by subcutaneous and intraperitoneal routes of inoculation. Albino rats did not develop antibody but rabbits developed haemagglutination inhibiting, neutralising and complement fixing antibodies. PMID- 2558277 TI - Using temperature-sensitive mutants of VSV to study membrane protein biogenesis. PMID- 2558279 TI - [The hemagglutinating properties and adsorption on erythrocytes of Coxsackie B-3 viruses and their selected bentonite variants]. AB - The paper is devoted to the study of the Coxsackie B-3 viruses and their bentonite variants passaged on the primary and transplanted cell cultures for their hemagglutinating (HA) properties and a degree of adsorption on erythrocytes. It is shown possible to detect the HA activity of the Coxsackie B-3 viruses which are passaged on the transplanted cell cultures: the variant Abent has the hemagglutinating activity relative to human and rabbit erythrocytes. The variant Abent+ of the Coxsackie B-3 virus passaged on the cell culture Vero is established not to be absorbed on erythrocytes. The variant Abent and the initial population of the Coxsackie B-3 virus are adsorbed on the human erythrocytes by 90-99%. Coxsackie B-3 virus and its bentonite variants reproduced on the primary cell culture of human embryonic fibroblasts and possessing the hemagglutinating activity are 100-1000 times better adsorbed on human erythrocytes than the viruses passaged in the transplanted culture. PMID- 2558280 TI - [A method for detecting the hemagglutinating activity of enteroviruses]. AB - Optimal conditions to reveal the hemagglutinating activity in the Coxsackie viruses of group B and hemagglutination test (HAT) performed by the micromethod are studied and determined. The titre of hemagglutinins revealed in HAT is established to depend on pH of the phosphate-buffer solution, concentration of human erythrocytes and their group attribution. The viruses passaged at the suboptimal temperature 33 degrees C are shown to possess the maximal hemagglutinating activity. PMID- 2558281 TI - [Basic and clinical research on intramural esophagography]. AB - Preoperative diagnosis of the penetration of esophageal cancer is important in its treatment. Lipiodol Ultra Fluid was injected into the oral side of the lesion, and after consideration of the state of its transfer linearly and histologically, the contrast medium had mostly transferred into the spaces between tissue. As a preoperative method of diagnosis, the ratio of the distance of transfer from the proximal edge of the tumor to the distal edge of the tumor in relation to the vertical length of the tumor was calculated and called the transfer ratio, and also (1) the distance from the assumed edge of tumor to the projecting portion in the inner cavity for the deepest part of the tumor was calculated, as was (2) the same calculation for the greatest penetration of contrast medium in the layer of adventitia, and the transfer ratio was multiplied by (2)/(1) and the result was called the section ratio. Using the transfer ratio and the section ratio, the preoperative depth of the invasion by the esophageal cancer was classified into four classifications: A0(sm), A0(pm), A1-2, and A3. The accuracy of diagnosis, comparing the preoperative diagnosis and the histological examination was: A0(sm): 100%, A0(pm): 75%, A1-2: 91.7%, A3: 80% and was 85.7% overall, showing very good result. PMID- 2558282 TI - Novel genes influencing the expression of the yellow locus and mdg4 (gypsy) in Drosophila melanogaster. AB - We used a system with a mobilized Stalker transposable element, sometimes in combination with P-M hybrid dysgenesis, in the search for new mutations interfering with the y2 mutation induced by mdg4 (gypsy) insertion into the yellow locus. A novel gene, modifier of mdg4, was detected in chromosome 3. The mutation mod(mdg4) either enhanced or suppressed phenotypic changes in different mutations induced by mdg4 insertions. Thus, mod(mdg4) seems to be involved in the control of mdg4 expression. Six other loci designated as enhancers of yellow were also detected. The e(y)n (with n from 1-6) mutations enhanced the expression of several y mutations induced by different insertions into the yellow locus. The major change is a damage of bristle and hair pigmentation which is not suppressed by su(Hw) mutations. On the other hand, e(y)n alleles do not interact with mdg4 induced mutations in other loci. All e(y)n genes are located in different regions of the X chromosome. One may speculate that e(y)n genes are involved in trans regulation of the yellow locus and possibly of some other loci. PMID- 2558283 TI - Molecular characterization of a specific p-nitrophenylphosphatase gene, PHO13, and its mapping by chromosome fragmentation in Saccharomyces cerevisiae. AB - The structural gene, PHO13, for the specific p-nitrophenyl phosphatase of Saccharomyces cerevisiae was cloned and its nucleotide sequence determined. The deduced PHO13 protein consists of 312 amino acids and its molecular weight is 34635. The disruption of the PHO13 gene produced no effect on cell growth, sporulation, or viability of ascospores. The PHO13 locus was mapped at 1.9 centimorgans from the HO locus on the left arm of chromosome IV. By chromosome fragmentation, the PHO13 locus was found to be located about 72 kb from the left hand telomere of chromosome IV and distal to the HO locus. PMID- 2558284 TI - Complete maps of IS1, IS2, IS3, IS4, IS5, IS30 and IS150 locations in Escherichia coli K12. AB - In this paper complete distribution maps are presented of the seven IS elements 1, 2, 3, 4, 5, 30 and 150. These maps were obtained during the construction of an almost complete restriction map of the Escherichia coli genome of K12 strain BHB2600. The positions of IS elements were correlated to this map. The distribution of integration sites of all IS types is nonrandom. Besides a large gap from 79 min to 96 min, there is a pronounced IS cluster at 6 min and another at 97 min, map locations that have low gene incidences on the classical map. One cluster coincides with a region of IS induced rearrangements. The IS distribution pattern was compared to patterns of strains W3110 and HB101. PMID- 2558285 TI - Analysis of the HO-cleaved MAT DNA intermediate generated during the mating type switch in the yeast Saccharomyces cerevisiae. AB - A galactose-inducible HO gene was used to induce mating type switching in heterothallic Saccharomyces cerevisiae cells arrested in G1, in rad52 mutants defective in DNA damage repair, and in cells lacking the donor cassettes. The HO cleaved MAT intermediate is stable over significant lengths of time, i.e. HO cleavage is not coupled to the subsequent gene conversion event. The in vivo cleavage site was mapped to single base resolution by primer extension experiments on total genomic DNA. Cells arrested in G1 with alpha-factor switched mating type thus demonstrating that switches can occur in the absence of replication of the genome. rad52 mutants did not produce MAT DNA of the opposite mating type indicating that the block is prior to the gene duplication stage of the switch. In strains in which the HM donor cassettes are deleted the cut MAT DNA was degraded after induction of the HO gene. PMID- 2558287 TI - Structural analysis of Doc transposable elements associated with mutations at the white and suppressor of forked loci of Drosophila melanogaster. AB - DNA sequences from two spontaneous mutations of Drosophila melanogaster associated with insertion of a Doc transposable element have been cloned. In white-one, the element is inserted in the white locus close to where transcription initiates. In a lethal allele of suppressor of forked, su(f)S2, the element is inserted within the transcription unit in the protein coding region. Four other Doc elements have been cloned from a wild-type strain. Doc is a member of the class of transposable elements known as retroposons, which includes the D. melanogaster F, G, Jockey, and I elements. There is no sequence homology between the ends of the Doc element. The 3' or right end terminates with a polyadenylation signal sequence followed by a stretch of oligo-A. The length of the oligo-A varies between elements, and a duplication of variable size is found as a direct repeat flanking inserted Doc elements. Members of the family are conserved at the 3' end, but may be truncated at the 5' or left end. These structural features suggest a mechanism of transposition via an RNA intermediate. PMID- 2558286 TI - Complete coding sequence of wheat phosphoribulokinase: developmental and light dependent expression of the mRNA. AB - We have isolated and sequenced a cDNA clone which contains the entire coding sequence of the precursor to a subunit of wheat phosphoribulokinase (PRKase). (The enzyme is a homodimer). The cDNA contains 1533 bp and has an open reading frame of 1212 nucleotides. This encodes a protein with an amino-terminal transit sequence of 53 amino acids, while the part that forms the mature protein contains 351 amino acids and has a molecular weight of 39,200 daltons. A comparison of the wheat amino acid sequence with that already known for the mature protein of spinach reveals that there are identical residues in 86% of the positions but their transit peptides differ substantially from one another. The mature wheat and spinach proteins are identical in a segment of over 50 amino acids near the amino-terminus which is the region believed to be involved in ATP binding and in regulation by light of the catalytic activity of the enzyme. We further demonstrate that the expression of PRKase mRNA in wheat leaves is regulated in a developmental, tissue-specific and light dependent manner. We also show that the light-induced increase in the steady-state levels of this mRNA is dependent on the developmental stage of the leaf. PMID- 2558288 TI - DNA superhelicity enhances the assembly of transcriptionally active chromatin in vitro. AB - Using an in vitro chromatin assembly system, we analyzed the influence of DNA superhelicity on the development of transcriptionally active minichromosomes. Plasmid DNA molecules containing either a Xenopus borealis 5S RNA gene or an X. laevis methionine tRNA gene were utilized as templates for the assembly of chromatin. Both plasmids were processed into active minichromosomes if introduced as supercoiled molecules into the extract (S-150). The degree of superhelicity is a determining factor in the assembly of active chromatin. Molecules containing varying superhelical densities were processed into minichromosomes with different transcriptional activities. The absence of supercoils leads to the assembly of chromatin with substantially lower transcriptional activity. Assembled minichromosomes are stable enough to be isolated by sucrose gradient centrifugation while retaining their transcriptional phenotype. The formation of nucleosomes with a periodic spacing occurred with the same efficiency and to the same degree regardless of the initial DNA topology. Hence, a determining factor in the development of transcriptionally active chromatin may be the initial superhelicity of the DNA molecule to which activator (trans-acting factors) or repressor (histones) proteins bind. Once the chromatin assembly process has begun, the transcriptional activity of the resulting minichromosome may already have been determined. PMID- 2558289 TI - Cauliflower mosaic virus promoters direct efficient expression of a bacterial G418 resistance gene in Schizosaccharomyces pombe. AB - A system is presented for transformation of the fission yeast Schizosaccharomyces pombe to resistance against the antibiotic G418. The bacterial resistance gene of the transposon Tn5 is expressed under the control of promoters and transcription terminators from cauliflower mosaic virus (CaMV). The promoter of the S. pombe alcohol dehydrogenase gene has also been used. Transformants can be selected directly on medium containing G418 (up to 1 mg/ml) due to inactivation of G418 by the Tn5 gene product, the aminoglycoside 3'-phosphotransferase (II). The plant viral promoter 35S confers higher resistance to G418 than the 19S promoter. This corresponds to the relative strengths of these promoters in plant cells. The strong plant promoter 35S yields resistance comparable to that obtained with the strong S. pombe promoter from the alcohol dehydrogenase gene. The constructions with the two plant promoters have been used on multicopy shuttle plasmids that replicate autonomously in S. pombe and Escherichia coli. In addition the 35S and the 19S constructions have been inserted into the S. pombe genome where they confer G418 resistance as single copy genes. Since vector sequences are excluded in this case, all the necessary signals for expression of G418 resistance are contained within the DNA fragments containing the plant promoters, the resistance gene and the plant terminators. This transformation system is independent of S. pombe mutants. It may be useful for the transformation of other lower eukaryotes. The activity of the CaMV promoters in S. pombe may be exploited for the expression of plant genes in fission yeast. PMID- 2558290 TI - [Characteristic features of the structure of co-integrating plasmids and simple insertions formed during transposition of the IS1 element in transposon Tn9']. AB - Earlier we have studied unstable dissociating IS1/Tn9'-mediated cointegrates between the plasmids pDK57 (pBR322::Tn9') and pRP3.1, a deletion derivative of RP1, and two types of such cointegrates containing three and four copies of IS1 were revealed. In the present paper we studied the structure of stable IS1/Tn9' mediates cointegrates and simple insertions formed by interaction between the plasmids pDK57 and pRP3.1 in the E. coli recA- cells. It was shown, that the stable cointegrates were formed by insertion of pDK57 in different loci of pRP3.1 and these cointegrates contain three copies of IS1, i.e. one copy of IS1 and a copy of Tn9' at the junction of the two replicons. The cointegrates are formed predominantly due to the activity of the left copy of Tn9', which occupies a proximal position in regard to the promoter of the cat gene. It was found that the integration of pDK57 into the kan gene region of pRP3.1 leading to the formation of the KmS cointegrates occurs only in one of the two possible orientations. Meanwhile the insertions of the transposon Tn9' into the kan region of pRP3.1 leading to simple insertions occurs in the orientation opposite to the orientation of the transposon in the KmS cointegrates. It is proposed that simple insertions are not the products of direct transposition of Tn9', but they are formed from unstable cointegrates under the action of IS1-specific resolvase. PMID- 2558291 TI - [Comparative analysis of sequences of protein D2 from photosystem 2 reaction center and various Ca 2+-binding proteins]. AB - The amino acid sequence of D2 protein was compared with those of calcium binding proteins and receptor for calcium channel blockers in connection with the data showing the participation of Ca2+ in photosystem 2 electron transport and the inhibition of this process by calmodulin antagonists, calcium channel blockers and local anesthetics. Protein D2 possesses a pattern analogous to the "EF-hand" sites of the calcium binding proteins. Comparison of the amino acid sequence of the calmodulin fragment binding the phenothiazine type calmodulin antagonists with the amino acid sequence of D2 protein and calcium channel protein revealed a high degree of sequence identity. Common structural features take place also between the membrane spanning segment III of D2 protein, which contains the tyrosine residue (161), responsible for ESR-signal IIS, and the membrane segment IVS5 of calcium channel protein. A model explaining the mechanism of calcium function in the oxygen-evolving system is proposed. PMID- 2558292 TI - [The family of env genes of avian retroviruses: molecular analysis of Rous sarcoma virus adapted to duck cells]. AB - For the elucidation of the molecular basis of RSV adaptation to conditionally permissive host from the genome library of duck embryo fibroblasts, transformed by Rous sarcoma virus in 30 passages on these cells, recombinant bacteriophages that include provirus sequences, were obtained. Complete and transformation defective proviruses were characterized, nucleotide sequences of their env-genes were compared with their counterparts the original RSV (Pr-RSV-C) and with viruses of other subgroups (A, B, D and E). The possible relation of the revealed changes in domains coding gp85 and gp37, with the changes of chicken RSV characteristics during adaptation to duck cells is discussed. PMID- 2558293 TI - [Cloning of tobacco DNA fragment with promoter properties in a transgenic plant]. AB - A selection system for isolating DNA sequences with transcription-promoting activity by their functioning in bacterial cells has been proposed. Tobacco nuclear DNA fragments were inserted in front of the promoterless neomycin 3' phosphotransferase II (NPT-II) gene and promoter-like sequences were identified by their ability to restore NTP-II activity in E. coli cells. One of these recombinant plasmids was introduced in tobacco protoplasts by direct gene transfer and transformed calli were isolated by kanamycin selection. The NTP-II expression in regenerated transgenic plants were highest in root, slightly lower in stem and were practically absent in leaf. Sequence analysis of cloned segment showed the presence of conserved sequences essential for promoter activity in eukaryotic cells. A transcription start site was observed by S1 mapping. The size of protected fragments corresponds to the initiation of transcription 176 and 179 base pairs upstream the initiation codon in tobacco and 75 base pairs in E. coli. PMID- 2558295 TI - Differentiated mammalian cell lines immortalized by temperature-sensitive tumor viruses. PMID- 2558294 TI - [Physico-chemical properties of low density lipoproteins isolated in an equilibrium density gradient]. AB - The tryptophanyls of total low density lipoproteins (LDL) (1.006-1.063 g/ml) from coronary heart disease (CHD) patients and subjects without CHD signs had different accessibility to fluorescence quenchers (I-and acrylamide). LDL were separated into subfractions in equilibrium density gradient. The coefficient of extinction , quantum yield and other spectral characteristics of LDL intrinsic fluorescence, rotational mobility of maleimide spin labels and fatty acid spin probe were different in LDL subfractions from healthy subjects. LDL subfractions with hydrated density 1.045-1.05 g/ml bound to B,E-receptors of cultured fibroblasts more effectively than did subfractions with density 1.01-1.03 g/ml. Structural differences of apo-B in the particles with different lipid to protein ratio are supposed. PMID- 2558296 TI - Regulation of human corticotropin-releasing hormone gene expression by 3',5' cyclic adenosine monophosphate in a transformed mouse corticotroph cell line. AB - In order to characterize potential mechanisms regulating the expression of the human CRH (hCRH) gene, an intact genomic fragment including 5'-flanking sequence of the hCRH gene was stably transfected into the mouse corticotroph AtT20 cell line. The exogenous hCRH gene was expressed at a high frequency with accurate and efficient transcription in transformed cells. Northern blot analysis revealed a single species of CRH mRNA of 1.6 kilobases which was identical in size to human placental CRH mRNA. S1 analysis demonstrated a single cap site in both placenta and transformed AtT20 cells, corresponding to a site 23 base pairs downstream of the TATA box. Treatment with 8-bromo cAMP and phorbol ester resulted in a dose dependent increase in CRH secretion during a 1-h incubation. Treatment with forskolin or 8-bromo-cAMP also produced a dose-dependent 4- to 10-fold increase in CRH mRNA levels, which was rapid (1 h) and sustained (6, 12, 24, and 48 h). These effects in a well characterized continuous cell culture system demonstrate pretranslational regulation of CRH expression by a cAMP-dependent pathway. PMID- 2558297 TI - A domain containing leucine-zipper-like motifs mediate novel in vivo interactions between the thyroid hormone and retinoic acid receptors. AB - The thyroid hormones and retinoic acid are potent modulators of differentiation, development, and gene expression. The transcriptional activities of these ligands are mediated by closely related nuclear receptors which bind and activate identical hormone responsive DNA elements. We noticed that a region within the ligand binding or E domain is well conserved between receptors for these hormones. This region contains hydrophobic heptad repeats that are structurally similar to the leucine-zipper dimerization domain. To study the function of this conserved domain, we examined the transcriptional responses of thyroid hormone receptor/c-erbA deletion mutants which lacked the heptad repeats. We previously reported that the chick c-erbA-alpha possesses hormone-independent (constitutive) activity in cells which express endogenous rat thyroid hormone receptor. We now demonstrate that this activity is abolished upon deletion of the conserved heptad repeats. This suggests that the heptad repeats mediate in vivo interactions between chick c-erbA and rat thyroid hormone receptors. To further test this hypothesis deletion mutants of chick c-erbA were constructed which contained all eight heptad repeats but which lacked the zinc-finger DNA binding domain. Although these mutants are transcriptionally inactive, they act in a dominant negative fashion to block trans-activation by both the chick c-erbA-alpha and the endogenous thyroid hormone and retinoic acid receptors. We suggest that the heptad repeats mediate the formation of inactive mutant/wild-type hetero-dimers. Dimer formation suggests a mechanism to account for the dominant-negative phenotypes displayed by nonhormone binding variants of c-erbA, the proto oncoprotein v-erbA and patients with the generalized thyroid hormone resistance syndrome. PMID- 2558298 TI - Glucocorticoid-regulated and constitutive trafficking of proteolytically processed cell surface-associated glycoproteins in wild type and variant rat hepatoma cells. AB - Glucocorticoids regulate the trafficking of mouse mammary tumor virus (MMTV) glycoproteins to the cell surface in the rat hepatoma cell line M1.54, but not in the immunoselected sorting variant CR4. To compare the localization of MMTV glycoproteins to another proteolytically processed glycoprotein, both wild type M1.54 cells and variant CR4 cells were transfected with a human insulin receptor (hIR) expression vector, pRSVhIR. The production of cell surface hIR was monitored in dexamethasone-treated and -untreated wild type M1.54 and variant CR4 cells by indirect immunofluorescence, direct plasma membrane immunoprecipitation, and by [125I] insulin binding. In both wild type and variant rat hepatoma cells, hIR were localized at the cell surface in the presence or in the absence of 1 microM dexamethasone. In contrast, the glucocorticoid-regulated trafficking of cell surface MMTV glycoproteins occurred only in wild type M1.54 cells. We conclude that the hIR, which undergoes posttranslational processing reactions similar to MMTV glycoproteins, does not require glucocorticoids to be transported to the plasma membrane and is representative of a subset of cell surface glycoproteins whose trafficking is constitutive in rat hepatoma cells. Thus, MMTV glycoproteins and hIR provide specific cell surface markers to characterize the glucocorticoid-regulated and constitutive sorting pathways. PMID- 2558299 TI - Regulation of transfected glycoprotein hormone alpha-gene expression in primary pituitary cell cultures. AB - Studies of gene regulation are greatly facilitated by the ability to transfect DNA into cultured cells. We examined a variety of transfection techniques to optimize transient expression of the human glycoprotein hormone alpha-gene in primary pituitary cells and subsequently investigated the regulation of alpha promoter transcription. Expression vectors driven by either the rous sarcoma virus-chloramphenicol acetyl transferase (RSVCAT) or the human alpha-gene (alpha CAT) promoters were transfected into cultures of dispersed female rat pituitary cells using calcium phosphate (CaPO4), diethylaminoethyl-dextran, lipofection, and electroporation procedures. CAT activity was optimal using the CaPO4 technique, resulting in 511 +/- 49% and 57 +/- 5% conversion/100 micrograms protein/4 h for RSVCAT and alpha CAT, respectively. Immunohistochemical analyses of alpha CAT expression using anti-CAT monoclonal antibodies demonstrated that the alpha-gene promoter is expressed in pituitary cells, predominantly if not exclusively, in gonadotropes and thyrotropes. Hormonal regulation of alpha promoter activity was assessed using both the CAT and the luciferase (LUC) reporter systems. alpha-Promoter activity was significantly (P less than 0.001) stimulated by 8-bromo-cAMP (217% increase), GnRH (75% increase), GnRH agonist analog (141% increase), and TRH (75% increase). The expression of control plasmids (RSVLUC, TKLUC, pOLUC) was not affected by treatment with these agents. We conclude that CaPO4-mediated transfection allows analyses of transient gene expression in primary pituitary cells. The alpha-promoter directs expression specifically in pituitary cells, predominantly gonadotropes and thyrotropes. alpha-Gene transcription is stimulated by GnRH, TRH, and 8-bromo-cAMP. PMID- 2558300 TI - Tumor-promoting phorbol ester and ras oncogene expression inhibit the glucocorticoid-dependent transcription from the mouse mammary tumor virus long terminal repeat. AB - Oncogene activation has been suggested to play some role in determining the hormone independency of tumors. In order to study the role of protein kinase C in mediating the inhibition of the glucocorticoid-dependent transcription from the Mouse Mammary Tumor Virus (MMTV)-Long Terminal Repeat induced by overexpressed activated ras oncogene, we studied the effects of protein kinase C activators [the tumor promoting phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA)] and inhibitors [1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7)] on the dexamethasone (DEX)-mediated activation of a MMTV-Long Terminal Repeat chloramphenicol acetyltransferase (pMMTV-CAT) chimeric reporter gene transiently transfected into NIH-3T3 cells and in Ha-ras-transformed fibroblasts (T24-NIH 3T3). TPA (30 ng/ml) together with DEX (0.1 microM) treatment of NIH-3T3 cells resulted in a significant decrease of CAT activity from pMMTV-CAT, compared to DEX treatment alone. The addition of H-7 (40 microM) was able to overcome the TPA induced inhibition of DEX-dependent transcription from pMMTV-CAT. DEX-dependent expression of pMMTV-CAT was significantly reduced in T24-NIH-3T3 with respect to wild-type NIH-3T3 cells. Treatment of T24-NIH-3T3 cells with either H-7 or TPA significantly enhanced or decreased, respectively, the DEX-dependent expression of pMMTV-CAT. TPA and/or H-7 did not affect CAT activity from either pMMTV-CAT in the absence of DEX or from CAT gene under the control of the SV40 promoter. Similar glucocorticoid receptor sites and binding affinities were observed in T24 NIH-3T3 or TPA-treated NIH-3T3 cells compared to wild-type untreated cells. Our data suggest that activation of PKC is involved in the reduced transcriptional regulatory activity of glucocorticoid hormone induced by overexpressed Ha-ras oncogene in NIH-3T3 fibroblasts. PMID- 2558301 TI - Activin stimulation of inhibin secretion and messenger RNA levels in cultured granulosa cells. AB - Recent reports suggest that activin (the dimer of inhibin beta subunits with FSH releasing activity) has specific receptors on ovarian granulosa cells. The present study examined the effects of purified porcine activin on inhibin secretion and mRNA levels in granulosa cells obtained from immature, estrogen treated rats. Cells were cultured for 48 h in culture media, or media containing FSH (10 ng/ml) and/or activin (30 ng/ml). Western blot analyses performed with affinity-purified antisera to inhibin alpha- and beta A-subunits revealed that treatment with either FSH or activin increased the secretion of inhibin alpha beta dimer (Mr 30,000), with a further increase after cotreatment. These results were confirmed by an inhibin alpha-subunit RIA, which revealed 7-, 14-, and 71 fold increases in the secretion of immunoreactive inhibin-alpha by activin, FSH, and activin plus FSH, respectively. TGF beta, a structural homolog of activin, also stimulated inhibin release, whereas follistatin was ineffective. Total RNA from cultured cells was hybridized with 32P-labeled inhibin alpha-subunit cRNA or beta-actin cDNA probes, and inhibin-alpha message levels were normalized with beta-actin mRNA levels. Northern blot analysis revealed that treatment with FSH and activin increased hybridization of a 1.5 kilobase (kb) message, corresponding to the inhibin alpha-subunit mRNA. Slot blot analyses indicated a 6- and 8-fold stimulation of inhibin alpha-subunit mRNA levels by FSH and activin, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558302 TI - Insulin-like growth factor-II (IGF-II): a potential autocrine/paracrine growth factor for human breast cancer acting via the IGF-I receptor. AB - Insulin-like growth factor-II (IGF-II) is a potent mitogen for several types of cultured cells and tissues. We have studied the interaction of IGF-II with a panel of cultured human breast cancer cell lines, examining the possibility that these cells synthesize and secrete IGF-II activity which could have autocrine/paracrine functions. Synthetic IGF-II was mitogenic in five of seven cell lines tested, including the estrogen receptor-positive lines MCF-7L, ZR75-1, and T47D and the estrogen receptor (ER)-negative lines Hs578T and MDA-231. IGF-II was slightly less potent than IGF-I in stimulating DNA synthesis in MCF-71 cells, an effect that paralleled its ability to compete for [125I]IGF-I binding in these cells. Affinity labeling studies revealed that IGF-II could also compete for binding to the 130,000 mol wt alpha-subunit of the IGF-I receptor. A monoclonal antibody to the IGF-I receptor inhibited the mitogenic effects of IGF-II in MCF 7L and MDA-231 cells, suggesting that this receptor mediates the growth effects of IGF-II in these breast cancer cells. Using a RIA and a RRA, IGF-II-like activity was detected in conditioned medium extracts processed to remove IGF binding proteins from several breast cancer cell lines, with the highest levels found in conditioned medium from MCF-7L and T47D cell lines. IGF-II mRNA transcripts in MCF-7L and T47D cells were identified by Northern blot analysis and were confirmed by RNase protection assay. IGF-II mRNA was increased by estrogen in MCF-7L cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558303 TI - Rapid induction of prostaglandin endoperoxide synthase in rat preovulatory follicles by luteinizing hormone and cAMP is blocked by inhibitors of transcription and translation. AB - Prostaglandin endoperoxide synthase (PGS) catalyzes the rate-limiting step in the synthesis of prostaglandins E and F2 alpha that are obligatory for ovulation. To understand the molecular mechanisms by which LH regulates the induction of PGS in rat preovulatory (PO) follicles, we established an in vitro system which mimics in vivo induction of the enzyme. We show that the rapid increase in PGS enzyme: 1) is stimulated by LH, FSH, and forskolin (cAMP) in a time- and dose-dependent manner that is distinct from changes in steroidogenic enzymes analyzed in the same follicles; 2) is unaltered by end products (PGE and PGF2 alpha) of the reaction or inhibitors (indomethacin) of enzyme activity; 3) is blocked by inhibitors of transcription (alpha-amanitin) and translation (cycloheximide) at a step distal to production of cAMP and activation of A-kinase. Analyses of PGS mRNA by Northern blots using a mouse PGS cDNA probe revealed a PGS transcript of 2.8 kilobases that was present but in low abundance in PO follicles and decreased rapidly (1-4 h) as a consequence of LH/human CG stimulation. This hormone-induced decrease in PGS mRNA appeared to be transient because PGS transcripts were present in corpora lutea at a level similar to that in PO follicles. These results raise the possibility that the marked increase in PGS enzyme in granulosa cells of PO follicles that occurs as a consequence of the LH/human CG surge may not involve a cAMP regulated increase in transcription of the PGS gene itself. Or, if increased transcription of PGS mRNA does occur, it is rapid and coupled with cotranslational degradation of the mRNA. Alternatively, transcriptional (alpha-amanitin-sensitive) regulation of a separate gene may be obligatory for increased translation of PGS mRNA or posttranslational modification (stabilization?) of the enzyme. In summary, the LH-stimulated appearance of PGS in granulosa cells of PO follicles before ovulation is mediated by cAMP in a complex manner involving transcriptional regulation (PGS gene?) and translational control of PGS mRNA. The transient appearance of the PGS enzyme represents a unique pattern of response by granulosa cells of PO follicles to LH/cAMP and thereby may involve novel intracellular factors and regulatory processes. PMID- 2558304 TI - Autophosphorylation of rat liver type II cAMP-dependent protein kinase. AB - A monoclonal antibody was prepared against the regulatory subunit (RII) of rat liver type II cAMP-dependent protein kinase. Autophosphorylated and nonphosphorylated RII in extracts from rat liver or hepatocytes were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and quantified by immunoblot analysis with this antibody. Under basal conditions, 90% of hepatocyte RII was in the phosphorylated form. Incubating hepatocytes with 8-bromo-cAMP and a phosphodiesterase inhibitor resulted in activation of cAMP-dependent protein kinase and glycogenolysis but did not affect phospho RII levels. RII phosphorylation was also unaffected by the inclusion of sufficient insulin to cause a decrease in cAMP-dependent protein kinase activity and glycogenolysis. The results indicate that unlike other cell types, dissociation of rat hepatocyte type II cAMP-dependent protein kinase does not result in dephosphorylation of RII. The biochemical basis for the apparent lack of RII dephosphorylation in intact hepatocytes was examined by comparison with smooth muscle where RII is rapidly dephosphorylated. Rat liver extract contained 4-fold less RII and had an 80-fold lower rate of dephosphorylation of endogenous RII compared to bovine smooth muscle extract. The differences in the rates of RII dephosphorylation in tissue extracts were not observed using purified RII from either tissue. These data suggested that the slow rate of RII dephosphorylation in rat hepatocytes is due to a difference in the susceptibility of endogenous rat liver RII to dephosphorylation rather than a difference in phosphatase activity. PMID- 2558305 TI - Molecular cloning of a mineralocorticoid (type I) receptor complementary DNA from rat hippocampus. AB - Rat brain expresses two types of corticosteroid-binding proteins. The type I receptor binds corticosterone with high affinity and is structurally related to the kidney mineralocorticoid receptor (MR), while the type II or classical glucocorticoid receptor binds corticosterone with lower affinity and displays an in vivo preference for dexamethasone. Here we describe the isolation and characterization of a cDNA coding for the MR, from a rat hippocampus cDNA library, by low stringency hybridization to radiolabeled human glucocorticoid receptor cDNA. The nucleotide and deduced amino acid sequence for rat hippocampal MR displays extensive homology to a MR cDNA isolated from human kidney, suggesting that they are orthologous genes. Southern analysis suggests that there is only one gene for the MR, and in vitro expression of the receptor generates a high affinity corticosterone-binding protein. These data provide evidence to support the contention that a single gene gives rise to the MR in renal tissues and type I receptors in the brain. PMID- 2558306 TI - Localization and regulation of glucocorticoid and mineralocorticoid receptor messenger RNAs in the hippocampal formation of the rat. AB - Messenger RNAs coding for glucocorticoid (GR) and mineralocorticoid (MR) receptor proteins were localized to discrete subfields of the hippocampal formation by in situ hybridization histochemistry, using cRNA probes of approximately equivalent specific activity. Both GR and MR mRNAs were present in all subfields examined; GR mRNA was of greatest abundance in CA1, while MR mRNA was most densely labeled in CA3. In all subfields examined, MR mRNA was considerably more abundant than GR mRNA. Removal of circulating glucocorticoids by adrenalectomy precipitated an up regulation of GR mRNA in subfields CA1-2 and the dentate gyrus, which was reversed by dexamethasone replacement. High doses of dexamethasone significantly down-regulated GR mRNA in CA3. In contrast, adrenalectomy produced significant up regulation of MR mRNA only in subfield CA1-2. The data indicate that steroid receptor mRNAs are differentially distributed in hippocampus, and that sensitivity to steroids occurs within defined structural domains of the hippocampal formation. PMID- 2558308 TI - Aldosterone induces a rapid increase in the rate of Na,K-ATPase gene transcription in cultured kidney cells. AB - Aldosterone (300 nM) induces a 4-fold increase over a 6-hr stimulation in A6 kidney cells from Xenopus laevis in the abundance of mRNA beta 1 and a 2-fold in that of mRNA alpha 1 coding for each Na,K-ATPase subunit, which is in agreement with a previous report. After a 3-hr stimulation already, aldosterone elicited a significant increase of mRNA beta 1 (2.51-fold +/- 0.47, n = 3, P less than 0.05) and a nonsignificant increase of mRNA alpha 1 (1.26-fold +/- 0.30, n = 3, NS). Increasing doses of cycloheximide up to 3 micrograms ml-1 led to 90% inhibition of protein synthesis, but failed to block the differential effect of aldosterone on mRNA abundance over a 6-h incubation period. The rate of transcription was measured by a nuclear run-on assay. The basal rate of mRNA alpha 1 transcription exceeded that of mRNA beta 1 by 2.8-fold. Aldosterone (300 nM) stimulated the transcription of the two subunit genes. Fifteen minutes after aldosterone addition there was a significant and parallel increase in the rate of transcription of the alpha 1 subunit (1.98-fold +/- 0.20, n = 3, P less than 0.02) and that of the beta 1 subunit (2.13 +/- 0.32, n = 3, P less than 0.04). After a 45-min stimulation period the transcription rate of the alpha 1 subunit remained at the level observed at 15 min (1.84-fold +/- 0.14, n = 4, P less than 0.01), while the transcription rate of the beta 1 subunit increased further (2.89 fold +/- 0.38, n = 4, P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558309 TI - Identification of a cDNA encoding a long form of prolactin receptor in human hepatoma and breast cancer cells. AB - Human PRL receptor cDNA clones from hepatoma (Hep G2) and breast cancer (T-47D) libraries were isolated by using a rat PRL receptor cDNA probe. The nucleotide sequence predicts a mature protein of 598 amino acids with a much longer cytoplasmic domain than the rat liver PRL receptor. Although this extended region has additional segments of localized sequence identity with the human GH receptor, there is no identity with any consensus sequences known to be involved in hormonal signal transduction. This cDNA will be a valuable tool to better understand the role of PRL in the development and growth of human breast cancer. PMID- 2558307 TI - Pituitary thyrotropin-releasing hormone receptors: local anesthetic effects on binding and responses. AB - Pharmacological agents are widely used to probe the mechanism of action of TRH. A number of these drugs behave as local anesthetics at high concentrations. The effect of local anesthetics on the binding of [3H]Me-TRH to specific receptors was studied using the GH4C1 line of rat pituitary tumor cells. [3H]Me-TRH binding was inhibited by classical local anesthetics with the order of potency (IC50 values): dibucaine (0.37 mM) greater than tetracaine (1.2 mM) greater than lidocaine (3.3 mM) greater than procaine and benzocaine (greater than 10 mM). IC50 values for other drugs with local anesthetic properties that inhibited [3H]Me-TRH were: 100 microM trifluoperazine, 100 microM imipramine, 170 microM chlorpromazine, 300 microM verapamil, and 700 microM propranolol. Inhibition by tetracaine and verapamil increased as the pH was raised from 6 to 8.5, indicating that the free base form of the amine drugs was the inhibitory species, and the local anesthetic effect was greater at 37 C than at 24 C or 0 C. [3H]Me-TRH binding to receptors in isolated membranes was inhibited to the same extent as binding to receptors on intact cells. Local anesthetics were 3- to 20-fold less potent at inhibiting [3H]Me-TRH to digitonin-solubilized receptors than binding to intact cells. In contrast, the potency of chlordiazepoxide, a putative TRH antagonist, to inhibit [3H]Me-TRH binding was equal using cells and solubilized receptors (IC50 = 10 microM). Local anesthetics inhibited TRH-stimulated PRL release and also inhibited basal PRL secretion and secretion stimulated by two nonhormonal secretagogues, (Bu)2cAMP and a phorbol ester.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558310 TI - AAEE case report #19: ischemic monomelic neuropathy. AB - A case study of a 50-year-old man who awakened from aortobifemoral bypass graft surgery with painful burning and numbness of the left foot is reported. Angiography showed embolic occlusion of the left popliteal artery proximal to the knee. Nerve conduction studies performed 1 year after occlusion showed loss or severe reduction of distal left sensory and motor amplitudes, and needle electrode examination showed fibrillation potentials in foot and distal leg muscles. The findings were felt to represent the result of acute ischemic nerve damage in the left leg. The clinical and electromyographic features of this entity will be described and comparisons will be drawn to experimental ischemic neuropathy and diabetic neuropathy. PMID- 2558311 TI - Thyrotoxic periodic paralysis and the sodium/potassium pump. AB - The hypothesis of altered Na+/K+ transport in thyrotoxic periodic paralysis (TPP) was tested in an investigation of the K+ influx into erythrocytes from two patients with episodes of thyrotoxic muscle weakness. A patient with primary hypokalemic periodic paralysis (HPP) and three healthy volunteers served as controls. The TPP patients were of Oriental and Caucasian origin and differed in their clinical symptoms. For the Caucasian patient, the Na+ content of the erythrocytes was twice the control, for the Oriental patient it was normal. The K+ dependence of the ouabain-inhibitable K+ influx (the pump action) was also abnormal in the Caucasian patient, the flux being 70% of control at 2 mM [K+]e and normal at 4 mM [K+]e. The K+ influx was normal in the Oriental patient. By contrast, the K+ leak of the cells was normal in the Caucasian and was increased in the Oriental patient. The pump/leak ratio was thus reduced in both TPP patients. All parameters investigated were normal in the patient with primary HPP. It is concluded that the ion transport systems of muscle may be altered in TPP, but that the patho-mechanism might be different in the rare Caucasian cases and the rather more common Oriental cases. PMID- 2558312 TI - Slow recovery of central axons in acrylamide neuropathy. AB - To study the differential vulnerability of central versus peripheral axons, we observed serial changes in conduction over comparable segments of central and peripheral axons of the primary sensory neuron in 17 rats with acrylamide neuropathy using somatosensory evoked potentials. Central conduction abnormalities persisted even after peripheral conduction and clinical abnormalities had recovered. Morphometric studies showed prompt restoration of the largest-diameter fibers in the peripheral nerve after clinical recovery but persistent or even more severe loss of large- and medium-sized fibers in the cervical gracile tract. This finding suggests that recovery from central peripheral distal axonopathy begins in the largest peripheral axons, perhaps even at the expense of central axons, and that clinical recovery can occur at a time when central conduction remains abnormal. The selective central axonopathy found in certain chronic degenerative disorders may be a consequence of this slow central recovery process associated with chronic or intermittent metabolic derangements. PMID- 2558313 TI - SFEMG improvement with remission in the cancer-associated Lambert-Eaton myasthenic syndrome. AB - In two patients with the Lambert-Eaton myasthenic syndrome (LEMS) and small-cell lung carcinoma (SLLC), therapy for LEMS was effective when combined with cancer therapy. The serial SFEMG tests showed a corresponding improvement with remission of the LEMS and proved to be a better and earlier indicator of the changing clinical status than the repetitive nerve stimulation test. Thus, we conclude that the SFEMG is useful in the follow-up evaluation of patients with this condition. PMID- 2558314 TI - Reversible axonal neuropathy from the treatment of AIDS and related disorders with 2',3'-dideoxycytidine (ddC). AB - A total of 20 patients with acquired immune deficiency syndrome (AIDS) or AIDS related complex (ARC) received the anti-retroviral drug 2',3',dideoxycytidine in a phase I study at doses ranging from 0.03 mg/kg every 8 hours to 0.25 mg/kg every 8 hours. Of the 11 patients who participated in the study for more than 5 weeks, 9 developed symptoms and signs of a mainly sensory painful neuropathy that was confirmed by electromyography to be mixed sensory and motor neuropathy of axonal type. The neuropathy which developed on dideoxycytidine occurred between 9 and 12 weeks of treatment. One patient, who had the drug stopped at 3 weeks owing to thrombocytopenia, developed a similar clinical picture of a sensory peripheral neuropathy after 2 weeks off dideoxycytidine. However, he did not have electromyographic evidence of a neuropathy, and he subsequently returned to normal clinically while taking a lower dose of dideoxycytidine in an alternating regimen. Five patients were withdrawn from the study because of the neuropathy. The pattern of this neuropathy was different from that of the slowly progressive painful neuropathy of AIDS, in that there was (1) a sudden onset of intense burning discomfort in both feet sparing the hands at about the tenth week (mean 10.4 weeks) of treatment, (2) there was motor involvement in some patients without progression, and (3) onset of the neuropathy was temporally related to the administration of dideoxycytidine and began to resolve 3-5 weeks after its discontinuation. We believe that dideoxycytidine can be an axonal toxin, especially when given in high dose continuous regimens.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558315 TI - Adynamia episodica hereditaria: what causes the weakness? AB - The cause of weakness was investigated in a patient with adynamia episodica hereditaria without myotonia. A pattern of exercise and rest produced episodes of hyperkalemic periodic paralysis. In addition, local muscle weakness was induced by forearm cooling. Investigations on isolated intercostal muscle demonstrated that a high potassium concentration in the bathing solution triggered a noninactivating membrane current causing depolarization of the muscle fibers. This current was carried by sodium as it could be inhibited by tetrodotoxin. The abnormal sodium conductance led to an increase of sodium within the fibers. This was demonstrated directly by intracellular recordings. Weakness induced by rest after exercise and cold-induced weakness appeared to have different pathomechanisms. In the cold, the muscle fibers retained a normal resting potential, but their excitability was reduced and their mechanical threshold was increased. These findings also provide evidence that the mechanism of cold induced weakness in adynamia episodica is distinctly different from the cold induced weakness that occurs in paramyotonia congenita. PMID- 2558316 TI - Ultrastructural localization of calcium binding sites on human muscle cell surface. AB - Calcium (Ca2+) is mainly bound to anionic phospholipids and to sialic acid at the cell surface. We studied the ultrastructural localization of these Ca2+ binding sites in normal human muscle fibers, using Polymyxin B as a marker for anionic phospholipids and the lectin Limulus Polyphemus as a probe for sialic acid. We found that anionic phospholipids have a patchy distribution along the muscle sarcolemma, with a preferential localization at the I band level and at the junction between the I and A band. Sialic acid has an uniform distribution along the muscle plasma membrane and basal lamina. Our observations suggest that the plasma membrane, basal lamina, and transverse tubular system play an important role in providing the negative charge of the human muscle cell surface and that these structures may be involved in the binding of calcium. PMID- 2558317 TI - Diagnostic use of magnetic nerve stimulator. PMID- 2558318 TI - Specific discontinuities in Leishmania tarentolae minicircles map within universally conserved sequence blocks. AB - Specific discontinuities remaining in the daughter strands of newly synthesized minicircles from Crithidia fasciculata and Trypanosoma equiperdum have been shown to lie within conserved sequence blocks (CSBs) present in minicircles from all species of trypanosomes. The presence of ribonucleotides at the 5' ends of the L strand in both cases suggests that the conserved sequences may constitute a replication origin. However, specific discontinuities have only been mapped in organisms having unusually homogeneous populations of minicircles. The specific nicks in minicircles from the heterogeneous population present in Leishmania tarentolae have now been mapped to predicted sites within the CSBs. These results are consistent with a universal mechanism of minicircle replication dictated by a conserve sequence motif present both in minicircles having a high degree of sequence heterogeneity as well as in relatively homogeneous populations. PMID- 2558319 TI - Presence of a fructose-2,6-bisphosphate-insensitive pyrophosphate: fructose-6 phosphate phosphotransferase in the anaerobic protozoa Tritrichomonas foetus, Trichomonas vaginalis and Isotricha prostoma. AB - Extracts of the anaerobic protozoa Tritrichomonas foetus, Trichomonas vaginalis and Isotricha prostoma contained a high activity (0.5-1 mumol min-1 (mg protein) 1) of pyrophosphate:fructose-6-phosphate phosphotransferase (PPi-PFK), but no detectable ATP: fructose-6-phosphate phosphotransferase. PPi-PFK from I. prostoma was purified close to homogeneity by adsorption on phospho-Ultrogel and elution with fructose-1,6-bisphosphate, and subsequent anion-exchange chromatography. The enzyme had an Mr of 95,000 as determined by gel filtration and consisted of subunits of Mr 48,000. PPi-PFK from I. prostoma and from T. foetus displayed hyperbolic kinetics with respect to their substrates and were not affected by fructose-2,6-bisphosphate. In sharp contrast with what has been found in other eukaryotes, no evidence could be found for the presence of fructose-2,6 bisphosphate in the two trichomonads, in I. prostoma and in Entamoeba histolytica. PMID- 2558320 TI - A kinetoplast DNA probe diagnostic for Leishmania major: sequence homologies between regions of Leishmania minicircles. AB - A restriction fragment from a cloned kinetoplast minicircle DNA has been shown to be diagnostic for Leishmania major. This 402-bp TaqI fragment has been used routinely (as a radiolabelled probe) to detect 10(4) parasites in simple dot blots, both experimentally and in epidemiological surveys. It positively identified all stocks of L. major tested (including all six known zymodemes) and showed very low homology to kinetoplast DNA (kDNA) and chromosomal DNA of Leishmania infantum and Leishmania tropica, two species commonly isolated from patients and wild hosts within foci of L. major in the Old World. DNA sequence analysis of a minicircle of L. major is reported for the first time, and it is demonstrated that this species shares with Leishmania aethiopica, Sauroleishmania tarentolae and several species of Trypanosoma a region of conserved sequence that is involved in DNA replication, a process that could present targets for selective chemotherapeutic attack. Sequence and restriction fragment analyses have indicated the difficulties of selecting species-specific sequences from kDNA which, even in the same parasite clone, contains several predominant minicircle classes, not all of which contain diagnostic sequences. PMID- 2558321 TI - [Tumor recurrence in the salvaged breast; a second chance?]. PMID- 2558324 TI - Naloxone inhibition of postprandial growth hormone releasing hormone-induced growth hormone release in obesity. AB - The effects of opiate receptor antagonist naloxone on growth hormone (GH) release after growth hormone-releasing hormone (GHRH) administration were investigated, before or after feeding, at 13.00 h, in 20 obese women and in 10 normal women. When GHRH was administered to obese women before a meal at lunch time, the mean peak plasma GH levels were very low, while plasma GH responses significantly increased after feeding. Naloxone, infused at a rate of 1.6 mg/h starting 1 h before GHRH administration (50 micrograms i.v. as a bolus), was capable of inhibiting GH release induced by administration of GHRH after feeding. On the contrary, naloxone did not induce significant variations on the fasting GHRH induced GH release. In normal women, naloxone did not significantly modify the GH response to GHRH, both before and after lunch. The inhibitory effect of naloxone indicates that in obese women there is an increased opioid activity, which could represent an abnormal response of the gastrointestinal tract to food ingestion. PMID- 2558325 TI - Immunoreactive delta sleep-inducing peptide secretion from mouse dissociated, anterior pituitary cells: regulation by corticotropin-releasing factor and arginine vasopressin. AB - Immunoreactive delta sleep-inducing peptide (IR-DSIP) has previously been localized to the ACTH/MSH cells of the human and porcine pituitary gland. In the present report, the distribution of IR-DSIP in the mouse pituitary gland was examined by immunocytochemistry. In this species, IR-DSIP was found to be co localized with thyroid-stimulating hormone (TSH) in anterior pituitary thyrotrophs and was also present in nerve fibers in the posterior and intermediate lobes. The effect of synthetic DSIP on IR-ACTH release from dissociated mouse anterior pituitary cells was also studied. DSIP (greater than or equal to 10(-9) M) inhibited both basal and CRF-induced IR-ACTH release from these cells. In addition, the effect of synthetic rat corticotropin-releasing factor (CRF) and arginine vasopressin (AVP) on IR-DSIP secretion was investigated. CRF and AVP at concentrations of 10(-11)-10(-7)M inhibited release of IR-DSIP from mouse anterior pituitary cells by 63%. When CRF and AVP (10(-10) 10(-7) M) were given concomitantly, the maximal inhibition of IR-DSIP release was observed at a concentration of 10(-8)M of CRF and AVP. However, these two peptides when given together showed no additive effect on IR-DSIP secretion. These findings suggest that during CRF induction of ACTH secretion from anterior pituitary corticotrophs, CRF may also act simultaneously to inhibit DSIP secretion from the thyrotrophs. PMID- 2558323 TI - The presence of ATP + ubiquitin-dependent proteinase and multicatalytic proteinase complex in bovine brain. AB - The presence of two distinct high-molecular-weight proteases with similar pH optima in the weakly alkaline region was shown in cytosol of the bovine brain cortex. They were separated by ammonium sulfate fractionation and each was further purified by DEAE-Sephacel, Sephacryl S-300, DEAE-Cibacron Blue 3GA agarose, heparin-agarose, and Sepharose 6B chromatography. The larger enzyme (Mr 1,400 kDa), which precipitates at 0-38% ammonium sulfate saturation, seems to be active in ATP + ubiquitin (Ub)-dependent proteolysis; it has low basal caseinolytic activity that is stimulated 3-fold by ATP, and when Ub is present ATP causes a 4.5-fold stimulation. A second proteinase was also found to be present (Mr 700 kDa) that precipitates at 38-80% ammonium sulfate saturation, is composed of multiple subunits ranging in Mr from 18 to 30 kDa, and degrades both protein and peptide substrates, demonstrating trypsin-, chymotrypsin- and cucumisin-like activities. Catalytic, biochemical, and immunological characteristics of this proteinase indicate that it is a multicatalytic proteinase complex (MPC), whose enzyme activity, in contrast to that of MPC from bovine pituitaries (1-3), is stimulated 1.7-fold by addition of ATP in the absence of ubiquitin at the early steps of purification; this property is lost during the course of further purification. Both proteinases are present in the nerve cells, since the primary chicken embryonic telencephalon neuronal cell culture extracts contain both ATP + Ub-dependent proteinase and MPC activities. PMID- 2558322 TI - Effects of taurine on cell morphology and expression of low-affinity GABA receptors in cultured cerebellar granule cells. AB - Effects of taurine and THIP were studied on the development of cultured cerebellar granule cells with regard to GABA receptor expression and morphological development. Culturing in the presence of taurine or THIP led to the formation of low affinity GABA receptors as revealed from Scatchard analysis of [3H]GABA binding. This formation of receptors was susceptible to inhibition upon culturing in the simultaneous presence of taurine and bicuculline demonstrating the involvement of the high affinity GABA receptors which are present on the cells regardless of the culture condition. Superfusion experiments on cells cultured under the different conditions demonstrated that the low affinity GABA receptors expressed after culturing in the presence of THIP or taurine mediated an inhibition by GABA of evoked transmitter release from the granule cells. Cells cultured in either plain culture media or in the presence of taurine were indistinguishable with respect to the number of neurite extending cells observed after 4 days in culture. In contrast, culturing in the presence of THIP increased the number of neurite extending cells by 8% relative to the controls. PMID- 2558326 TI - Neuroendocrine, sympathetic and metabolic responses induced by interleukin-1. AB - Effects on turnover of vasopressin (AVP) in the hypothalamus and on secretion of pituitary hormones, catecholamines and insulin after intraperitoneal injection of recombinant interleukin-1 (beta) (IL-1) were investigated in male wistar rats. Intraperitoneal administration of IL-1 in a dose (1 microgram) that maximally activated pituitary-adrenal activity failed to alter plasma concentrations of prolactin, luteinizing hormone and melanocyte-stimulating hormone. Rats chronically cannulated in the right jugular veins showed a time-related increase in plasma corticosterone concentrations in response to intraperitoneal administration of IL-1 that lasted up to 4 h. In the same rats, plasma epinephrine (E) and norepinephrine (NE) concentrations were only slightly elevated (2-fold increase) at 30 min and at 1 h after IL-1 administration. Unlike in endotoxin-resistant C3H/HeJ mice, where IL-1 induces hypoglycemia, IL-1 did not affect plasma concentrations of glucose and insulin in Wistar rats. In the zona externa of the median eminence, IL-1 stimulated corticotropin-releasing factor (CRF) turnover at an approximate rate of 15%/h, but did not cause a concomitant change in AVP turnover as can be observed after insulin-induced hypoglycemia. Since half of the hypothalamic CRF neurons have been shown to costore AVP, the data favor the view of a selective effect of IL-1 on a subtype of CRF neurons. We conclude that pituitary-adrenal activation in response to Il-1 is caused by CRF secretion from a subtype of CRF neurons (not storing AVP) in the rat hypothalamus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558327 TI - Immunohistochemical evidence that most rat corticotrophs contain beta-adrenergic receptors. AB - Cells bearing beta-adrenergic receptors (beta-AdRs) were found in the anterior lobe of the rat pituitary gland using antiserum generated against affinity purified beta 2-AdRs, which also cross-reacts with beta 1-AdRs. beta-AdR-positive cells were sparsely distributed in the anterior pituitary gland. The ultrastructure of these cells was similar to that of corticotrophs. Subsequent analysis of adjacent serial sections indicated that 75% of the ACTH-positive cells were beta-AdR-positive. Our findings demonstrate the presence of beta-AdRs on pituitary corticotrophs, which provides an anatomical basis for the direct regulation of corticotroph secretions by beta-adrenergic agents. PMID- 2558328 TI - Neonatal handling alters adrenocortical negative feedback sensitivity and hippocampal type II glucocorticoid receptor binding in the rat. AB - Adult rats handled (H) daily for the first 3 weeks of life show a dramatically altered adrenocortical response to stress. We found that H animals secreted less ACTH and corticosterone (B) during and following the termination of stress than did nonhandled (NH) controls. In contrast, H and NH animals did not differ in basal B secretion at any point in the diurnal cycle, nor in adrenocortical responses to exogenously administered oCRF or ACTH. Moreover, the clearance rate for B was similar in H and NH animals. H animals were more sensitive than NH animals to the inhibitory effects of either B or dexamethasone on stress-induced adrenocortical activity. In a dose-response study, both glucocorticoids administered 3 h prior to testing suppressed the adrenocortical response to a 20 min restraint stress to a greater extent in the H animals. Handling increased type II, glucocorticoid receptor binding capacity in the hippocampus of adult animals (approximately 50% increase in capacity, with no change in affinity). There were no handling-induced changes in type II receptor binding capacity in the hypothalamus or pituitary, nor in type I receptor binding capacity in the hippocampus. Following chronic (5 mg/kg/day) treatment with B, hippocampal type II receptor binding capacity was significantly reduced in the B-treated H animals, compared with saline-treated H animals, and indistinguishable from saline-treated NH animals. Down-regulated H animals, like NH animals, hypersecreted B following the termination of stress in comparison to the saline treated H animals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558330 TI - Seizure-triggering mechanisms in the kindling model of epilepsy: collapse of GABA mediated inhibition and activation of NMDA receptors. AB - Our recent studies on seizure-triggering mechanisms in the kindling model of epilepsy are reviewed. Electroencephalographic (EEG) events during kindling inducing tetanic stimulation from the site of stimulation were recorded, with an emphasis on EEG suppression and rhythmic synchronous discharge. From electrophysiological and pharmacological analyses of these events, it is hypothesized that activation and subsequent collapse of GABA-A-mediated inhibition is an essential precondition in the initiation of kindled seizures. The excitatory role of NMDA receptors in kindling were also investigated by examining the effects of a noncompetitive antagonist of NMDA receptors (MK-801) on amygdala kindling and hippocampal long-term potentiation (LTP). The results indicate that activation of NMDA receptor complex combined with the collapse of GABA-A-mediated inhibition may be critical for kindling development. PMID- 2558329 TI - [Comparison of the efficacy of monotherapy with a beta-blocker, a diuretic, and ACE inhibitors in the control of blood pressure during stress]. AB - In order to compare the efficacy of beta-blocking, diuretics and ACE-inhibiting monotherapy in controlling the blood pressure increase to stress, a study was conducted on 30 subjects (10 treated with atenolol, 10 with hydrochlorothiazide/amiloride combination, 10 with enalapril) with mild or moderate essential hypertension whose resting blood pressures were normalised by therapy. In the 3 groups of subjects blood pressure values at rest, during mental stress, static and dynamic exercise did not significantly differ before antihypertensive therapy. Atenolol and enalapril significantly reduced systolic and diastolic pressure below pretreatment values throughout and immediately after each test, differing from diuretic therapy which did not show any significant reduction in diastolic rises at the peak of hand-grip or in both systolic and diastolic pressures at the highest work-loads during dynamic exercise. In the recovery period of the exercise cycle test diuretics also produced a later normalisation of diastolic pressure. In conclusion, beta-blockers and ACE inhibitors seem to be more effective than diuretics in the control of the blood pressure response to stress in hypertensive patients, suggesting that these drugs are the first choice treatment of mild to moderate hypertension. PMID- 2558331 TI - TCP binding: a tool for studying NMDA receptor-mediated neurotransmission in kindling. AB - Findings from numerous pharmacological and electrophysiological studies have uniquely implicated the N-methyl-D-aspartate (NMDA) receptor in kindling. Recent findings indicate that this receptor is regulated by ligands acting at both amino acid (NMDA and glycine) and ion (Zn++ and Mg++) binding sites. To examine the role of the NMDA receptor in kindling it will be necessary to understand how ligands for these different binding sites interact to control activation of the NMDA receptor. To this end we examined a biochemical tool for measuring opening of the NMDA receptor-gated ion channel (NMDA channel). [3H]N-(1-[thienyl] cyclohexyl)piperidine (TCP) binding to brain membranes is stimulated by NMDA and glycine receptor agonists. We have shown that NMDA and glycine increase TCP binding by increasing the access of TCP to its site. Moreover, the pharmacology of the NMDA and glycine binding sites regulating TCP binding is identical to that of the sites regulating NMDA evoked currents. These findings strongly suggest that glycine and NMDA regulate TCP binding by increasing the opening of the NMDA channel. That is NMDA and glycine increase the overall time that the channel is open thereby increasing the time available for TCP to diffuse to its binding site. These findings support the use of TCP binding (association rate) as a marker of channel opening and thereby permit measurement of NMDA receptor activation and ligand binding under identical conditions. This will allow direct testing the hypothesis that an alteration in the NMDA receptor/channel complex itself underlies the increased seizure response of kindled animals. PMID- 2558332 TI - Pyriform cortex involvement in kindling. AB - Evidence suggests that the pyriform cortex (PC) may play an important role in the genesis, if not the maintenance, of secondarily generalized limbic kindled seizures. For example, it has been shown that the fastest rates of kindling are observed from structures most directly related to the PC, and that the latter develops epileptic burst responses before all other structures, independent of the kindling site. This seizure sensitivity of the PC is reflected additionally in its inevitable loss in the face of protracted seizure, i.e., status epilepticus. In conclusion we briefly review our electrophysiological data from amygdala-pyriform cortex slices which show that the PC and overlying perirhinal cortex (PRC) possess a strong disposition for developing spontaneous rhythmic burst discharges. Further it was observed that the PRC response always led the PC event in control tissue, but rarely in kindled tissue. This suggests a functional change in the relationship between these two areas as a result of amygdala kindling. The significance of this alteration is yet undetermined. PMID- 2558333 TI - [Immunomodulation by leukocytolysis. On the significance of lectin-polysaccharide complexes]. AB - Polymorphonuclear granulocytes (PMNs) are thought to function only as phagocytes. The ability of PMNs for a physiologically lysis has not been noticed so far. Therefore, PMNs are not only a clamp between the specific and unspecific immune system but an important regulator of the homoeostasis. All immune modifiers therapeutically used seem to stimulate the physiologically lysis of PMNs. This has been shown with mistletoe lectins. Most remarkable are findings that show lectins bound to polysaccharides drastically rise the ability of PMNs to undergo lysis. PMID- 2558334 TI - The chromosome 11 region flanking the t(11;14) breakpoint in human T-ALL is deleted in Wilms' tumor hybrids. AB - Human chromosomal band 11p13 has been implicated in T cell malignancies carrying t(11;14)(p13;q11) reciprocal translocations and has also been associated with Wilms' tumor and aniridia in a mechanism characterized by overlapping hemizygous constitutional deletions spanning this region. Using probes derived from the T cell receptor delta gene, we have cloned the chromosomal breakpoint in an acute T cell leukemia (T-ALL). Southern blotting analyses of mouse-human somatic cell hybrids from this human T-ALL sample and Chinese hamster-human somatic cell hybrids derived from Wilms' tumor lines have indicated that the 11p13 locus, tcl 2, juxtaposed to the TCR (T cell receptor) delta locus in T cell leukemia, is within the constitutional deletion of two Wilms' tumor-aniridia cases. PMID- 2558335 TI - Renin-promoter SV40 large T-antigen transgenes induce tumors irrespective of normal cellular expression of renin genes. AB - Chimeric genes containing the 5'-flanking regions of the mouse renin genes, Ren1 and Ren2, associated with the early region of the simian virus 40 (SV40) were constructed. The two recombinant genes which contain, respectively, 0.45- and 2.5 kb the Ren1 and Ren2 5'-flanking sequences, named Ren1Tag and Ren2Tag, were microinjected into fertilized eggs. Tumors arose after a latency of 5-9 months in mouse lines harboring these hybrid genes except for one, in which a different and earlier pathology was observed (peripheral neuropathies). Most of the pathologies developed by these transgenic mice reflect the tumorigenic spectrum of the SV40 early region gene (choroid plexus, kidney, intestinal tumors, and peripheral neuropathies). None of these tumors arose from renin-producing cells nor produced renin. As suggested by the tumor pathology, the expression of the SV40 large T antigen did not follow the normal expression of the Ren1 and the Ren2 genes since SV40 large T-antigen mRNA was found in tissues which normally do not express renin. PMID- 2558336 TI - Expression level of the retinoblastoma susceptibility gene is elevated in simian virus 40-transformed cells. AB - We examined the level of mRNA of the retinoblastoma-susceptibility gene (Rb gene) in simian virus 40 (SV40)-transformed primate cells. The mRNA level of Rb gene was elevated in SV40-transformed cells compared with that in untransformed parental cells. The level of Rb message was also increased by SV40 infection on monkey kidney CV-1 cells. The level of Rb gene mRNA in SV40-transformed or SV40 infected cells was about fourfold higher than that in controls. These observations suggest that Rb gene is one of the cellular genes which can be elevated by SV40 directly or indirectly. PMID- 2558337 TI - Study on genetic polymorphism of Leishmania infantum through the analysis of restriction enzyme digestion patterns of kinetoplast DNA. AB - Twenty-nine Leishmania infantum strains characterized by different host source, tropism and belonging to 6 zymodemes, were examined by restriction enzyme analysis of kinetoplast DNA (kDNA) using 15 endonucleases. The enzymes which produced only one fragment revealed full identity between all the strains examined, while those producing many bands gave different electrophoretic patterns. They were interpreted with the aid of numerical analyses (cluster and multifactorial analysis). The results show a cline of genetic variability among the strains, the highest similarity being observed between most of the viscerotropic strains isolated from man, dog, black rat and sandflies. The strain agents of human cutaneous leishmaniasis show a varying degree of genetic divergence from this group, which appears more evident when characters from isoenzymes are considered. PMID- 2558338 TI - A preliminary assessment of the feasibility of evaluating promising antifilarials in vitro against adult Onchocerca volvulus. AB - The suitability of motility indices and tetrazolium-based colorimetric assays for the determination of the viability of adult Onchocerca volvulus after in vitro exposure to potential macrofilaricides has been examined. Experimentation showed that both techniques could be applied to adult O. volvulus, although the variability between individual worms necessitated the use of large experimental groups. The potential of using cut anterior tips of female O. volvulus for screening was also investigated. These were shown to give reasonably consistent motility indices, and drug effects were discernible even after 72 h in vitro culture. Application of these viability criteria to studies on the short-term in vitro survival of intact male and female O. volvulus incubated in Eagles MEM plus serum, under 5% CO2 in air, showed this medium to be suboptimal with a greater than 50% loss of worm viability within 144 h of nodulectomy. Males isolated by the collagenase technique were shown to be significantly less viable than dissected males, by both motility indices and tetrazolium reduction. The results highlight the need to use either dissected males, or in the case of females, the need to minimize exposure to collagenase solution. A possible mechanism for selecting a more uniformly viable female worm population is discussed. Examination of the in vitro effects of CGP 20376 using these viability criteria/assay systems showed some delayed suppression of worm motility, but after 120 h in vitro CGP 20376 was not macrofilaricidal against male or female O. volvulus. Male worms were also implanted subcutaneously into gerbils.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558339 TI - [Production, purification and evaluation of human cytomegalovirus 66 kD structural antigen for antibody detection by ELISA]. AB - An ELISA method using human CMV major polypeptide (66 kD) purified by electroelution was used to detect IgG and IgM antibodies and was compared with a conventional ELISA using the whole virus. The ELISA-66 kD showed a good sensitivity (98.2%) and had a better specificity than the ELISA-whole virus (97.5% versus 80.3%). Cross reactions observed, particularly with sera containing anti-EBV antibodies, were ruled out. PMID- 2558340 TI - [Antioncogenes]. PMID- 2558341 TI - [Papillomavirus and cancer of the uterine cervix. The virologist's point of view]. PMID- 2558342 TI - [Papillomavirus and cancer of the uterine cervix. The clinician's point of view]. PMID- 2558343 TI - [Therapeutic intensification with bone marrow autografting in cancer in children]. PMID- 2558344 TI - [Genetic factors in colorectal cancer]. PMID- 2558345 TI - [Insulinomas in children (characteristics of clinical picture and surgical treatment)]. AB - The paper treats of the main characteristics of the clinical picture and diagnosis of insulinoma in children as compared to adults. Seven children were operated on for insulinomas at the Surgery Department of the All-Union Research Endocrinology Center of the USSR AMS. The clinical course of insulinomas in children was characterized by a short-term disease history, the lack of overweight, and the convulsive syndrome as the leading symptom of hypoglycemia. As to the diagnostic tests, the fasting test appeared not desirable in the majority of children because of the low blood content of glucose in the morning hours and development of a marked hypoglycemic attack. Examination of immunoreactive insulin was not so indicative as in adults. During convulsions, electroencephalography in children was not feasible. Visceral arteriography turned out a reliable method of topical diagnosis of insulinoma in children. Tumor was most frequently located in the tail of the pancreas. The postoperative period in children ran a more favourable course than in adults. No clinical signs of pancreatitis were recorded. According to follow-up studies, the patients did not show any clinical or biochemical signs of hypoglycemia. Histological examination demonstrated that children had mainly neoplasms from beta-cells of islets of Langerhans. It is suggested that children have very low power to adjust themselves to acute and chronic hypoglycemia as compared to adults. PMID- 2558346 TI - [Precocious puberty syndrome in a body with hormone-producing tumor of the liver]. PMID- 2558347 TI - Alkylated cAMP derivatives: selective synthesis and biological activities. AB - Many alkylated cAMPs have been prepared and tested for their antitumor and cardiac activities. Treatment of cAMP with several bases and alkyl bromides gave alkyltriesters of cAMP (2), N6,N6,2'-O-trialkyl cAMPs (3), N6,2'-O-dialkyl cAMPs (4) and 2'-O-monoalkyl cAMPs (5) in one step. N6,N6-dialkyl cAMPs (6) were prepared from 2'-O-tosyl cAMP by the similar alkylation, followed by detosylation. Synthesis of N6-monoalkyl cAMPs (7) was achieved by the reductive alkylation of cAMP with aldehydes in one step. Alkyl triesters of cAMP exhibited antitumor activities against P815 cells. N6-mono and N6,N6-dialkyl cAMPs showed significant cardiac activities. PMID- 2558348 TI - Synthesis of imidazo[4,5-d][1,3]thiazine ribosides and related compounds of biological interest. AB - Several tri-O -acetyl-1-beta-D -ribofuranosyl- and 2 acetoxyethoxymethylimidazo[4,5-d][1,3]thiazine derivatives 5-10 were synthesized from imidazo[4,5-d][1,3]thiazine 4 and tetra-O -acetyl-beta-D -ribofuranose or 2 acetoxyethoxymethyl acetate by fusion method. In each case, depending on the substituent on position 5 of thiazine skeleton, only N3 isomer or both N1 and N3 isomers (in different ratios) were obtained. PMID- 2558349 TI - Effect of pressure on plant endonuclease reactions. AB - Effect of pressure on plant endonucleases, nuclease P1 from penicillium and an endonuclease from potato, was investigated especially on the influence on phosphomonoesterase and phosphodiesterase activities shown on substrates of XpYp type, as well as their intrinsic pressure-stability. The potato enzyme was found to be far less pressure-sensitive in both senses. PMID- 2558350 TI - Synthesis and selected properties of oligonucleotidyl-(Pm----O)-amino acids. AB - N-Cbz-Ser(OMe)-(Pm----O)-d(TpT) was synthesized as a diastereomeric mixture of approx. (1:1) by reacting d(TpT) with 2,4,6-triisopropylbenzenesulfonyl chloride and N-Cbz-Ser-OMe. The phosphoesteric bond of N-Cbz-Ser (OMe)-(Pm----O)-d(TpT) was found to be stable in acid (1 N HCl, 1 h, 37 degrees C), but labile in alkaline solution (0.1-1 N NaOH, 1 h, 37 degrees C). The products of alkaline hydrolysis were determined to be d(TpT) and the amino acid derivatives. Furthermore, the phosphotriester N-Cbz-Ser(OMe)-(Pm----O)-d(TpT) was more labile than the diesteric analogue N-Cbz-Ser (OMe)-pdT. The internucleotide phosphotriester linkage of N-Cbz-Ser (OMe)-(Pm----O)-d(TpT) was also found to be resistant to enzymatic digestion with spleen and snake venom phosphodiesterases. PMID- 2558351 TI - Syntheses and properties of oligodeoxyribonucleotide analogues having diastereochemically pure phosphoramidate linkages. AB - The oligothymidilate analogues, having stereoregular and alternative phosphormorpholidate/phosphodiester backbone, were synthesized from the diastereochemically pure dimer blocks by phosphorbisamidite method. The phosphormorpholidate linkage of the oligothymidilate analogues were resistant to snake venom phosphodiesterase, and no cleavage of the phosphodiester linkage of the analogues were slowly cleaved by the nuclease was slow. The abilities of isomers to form the complexes with poly(dA) greatly depended on their structures. PMID- 2558352 TI - Streptomyces ATP nucleotide 3'-pyrophosphokinase is also an ApppA nucleotide 2',3'-cyclic phosphokinase. PMID- 2558353 TI - Three-dimensional structure of ribonuclease Ms*3'-guanylic acid complex at 2.5 A. PMID- 2558354 TI - ESR behaviour of spin-labelled oligoDNAs. AB - Oligodeoxyribonucleotides (oligoDNAs) were spin-labelled at the 5'-end internucleotide linkage, directly or through the spacers of different lengths, with TEMPO and the labelled oligoDNAs were explored for the ESR spectroscopical behaviour in solution. Oxidation of H-phosphonate intermediates in CCl4 solutions of TEMPO and diamines was taken for the introduction of TEMPO and spacers, respectively. ESR lines of TEMPO attached to oligoDNAs, directly or through spacers, were already broadened compared to those of free TEMPO, and the broadening decreased with increasing chain length of the spacer and increased with increase in the chain length of the oligoDNAs. The ESR lines of the oligoDNA labelled directly were further broadened in the presence of a DNA complementary to the labelled oligoDNA. In addition, the broadened lines were further broadened in the presence of a larger DNA. This observation implies that spin labelling at the internucleotide linkage is useful in the DNA probe method to avoid B/F separation. PMID- 2558355 TI - 8-Methyladenosine-substituted analogs of 2-5A: synthesis and properties. AB - Four 8-methyladenosine-substituted analogs of 2-5A were obtained by a synthesis which involved a modification of the lead ion-catalyzed ligation reaction. Substituent of an 8-methyladenosine residue at the third position (2'-terminal) of the oligonucleotides increased the stability to snake venom phosphodiesterase digestion. PMID- 2558356 TI - Measurement of the proliferative status of colonic epithelium as a risk marker for colon carcinogenesis: effect of bile acid and dietary fiber. AB - The proliferative status of mouse colonic epithelium, as affected by dietary fibers with or without cholic acid (CA), was studied by autoradiography and the metaphase arrest technique. In the first study, groups of mice were fed natural ingredient (laboratory chow) or semisynthetic diets containing 0% (control) or 0.2% (test) CA. After the mice were fed two weeks, the effect of CA was significantly more pronounced in the semisynthetic diet group than in the natural ingredient diet group with respect to labeled cells/crypt section (7.8 +/- 0.8 vs. 2.9 +/- 0.4) and mitotic figure (MF)/crypt section (3.0 +/- 0.5 vs. 1.8 +/- 0.2). In the second study, diets formulated to contain 5 or 10% cellulose (C), pectin (P), or wheat bran (WB) with or without CA (0.2%) were fed to animals for two weeks and colonic proliferative indices were measured. When compared with 5% C group, the 10% WB group exhibited lower labeling index (LI) values (4.2 +/- 0.5 vs. 6.4 +/- 1.0) and the 10% P group exhibited higher LI values (10.0 +/- 1.1 vs. 6.4 +/- 1.0). CA-induced increases in the LI and MF values responded independently in some cases to dietary fiber. Among the CA-treated groups, only the 10% P diet resulted in lower LI when compared with the 5% C group (p less than 0.05) (7.4 +/- 0.8 vs. 12.5 +/- 2.8) but had no effect on MF/crypt section. However, the 5 or 10% WB diet resulted in lower MF values (1.7 +/- 0.2 and 1.8 +/ 0.6 vs. 2.6 +/- 0.3). A long-term feeding study comparing 10% P with 10% C diets also demonstrated that the LI was elevated in the 10% P group without any effect on the mitotic activity of the colonic epithelium. This paradoxical finding suggests that the value of the LI and/or mitotic index as a risk marker of colon carcinogenesis should be further investigated. PMID- 2558357 TI - Levels of vitamin A and cellular retinol binding protein in human hepatocellular carcinoma and adjacent normal tissue. AB - The levels of vitamin A (retinol) and vitamin E were measured in the blood, in tissues of human hepatocellular carcinoma (HCC), and in adjacent liver parenchyma. The median values of vitamin A were 11.5 micrograms/g (ranging 0-82.5 micrograms/g) in HCC and 52.1 micrograms/g (ranging 0.4-895.2 micrograms/g) in normal liver tissues; the difference was statistically significant (p less than 0.05). By contrast, there was no significant difference in vitamin E levels between the two tissues. Although the levels of vitamin A were significantly lower in HCC in 10 patients, no significant difference was noted in the cellular retinol binding protein levels in the normal and malignant tissues. These results suggest that the decreased levels of vitamin A in HCC are not due to altered cellular retinol binding protein levels in tumors and the different vitamin A blood supply system. We conclude that either the decreased uptake of vitamin A, but not vitamin E, by HCC cells or the lack of vitamin A-storing cells in tumors might be responsible for the low levels of vitamin A in HCC. PMID- 2558358 TI - The prognostic significance of basic anthropometric data in children with advanced solid tumors. AB - In pediatric cancer patients, malnutrition is commonly observed. This may represent the metabolic effect of the primary disease or it may be a consequence of multimodal therapy. This report evaluates the efficacy of using basic anthropometric measurements to predict morbidity during therapy. Twenty children with Wilms' tumor (Stage III, IV, and V) or neuroblastoma (Stage IV) diagnosed at Children's Hospital (Columbus, OH) between January 1983 and December 1985 were evaluated. When compared with the Wilms' tumor patients, the children with neuroblastoma had a significantly lower weight for age at diagnosis. At the completion of therapy, both weight-for-height and weight-for-age measurements were statistically lower in the neuroblastoma group (p less than 0.05). Significant differences were observed between the neuroblastoma and Wilms' tumor patients in the morbidity reported during therapy. Children with neuroblastoma had more frequent hospital admissions, spent a much greater proportion of their treatment time as hospital inpatients, experienced longer delays in therapy, and sustained many more complications. Each of the anthropometric indices was evaluated as a predictor of the complications observed during treatment. In the Wilms' tumor group, the patients with lower weight-for-height percentiles had an increased incidence of incomplete drug infusions, many more complications, more frequent hospital admissions, and an increase in the percentage of time spent as hospital inpatients. In the neuroblastoma group, the anthropometric measurements had no correlation with the subsequent development of complications. Nutritional staging based on anthropometric measurements recorded at diagnosis may be useful in predicting an increased risk of morbidity during therapy in children with Wilms' tumor. PMID- 2558359 TI - Supraglottitis in three young infants. AB - Three case histories of young infants with supraglottitis are presented. At this age, supraglottitis rarely occurs. Atypical features, as compared to older children, include a viral prodrome, lack of fever, stomatitis, and negative blood cultures. Although viral supraglottitis has been previously reported, this is the first report of epiglottitis associated with parainfluenza virus. PMID- 2558360 TI - Effects of calcitonin gene-related peptide on airway epithelial functions in dogs. AB - We studied the effects of calcitonin gene-related peptide (CGRP) on ciliary beat frequency (CBF) and electrical properties of canine tracheal epithelium by a photoelectric method and Ussing's short-circuit technique, respectively. CGRP dose dependently increased CBF, an effect that was accompanied by elevation of intracellular cyclic AMP but not affected by blockade of either Ca2+-influx or arachidonic acid metabolism. In contrast, CGRP elicited only a small and transient increase in short-circuit current without significant alterations in transepithelial potential difference or tissue conductance. These results suggest that CGRP may play a role in regulating airway mucociliary transport function. PMID- 2558361 TI - Recovery of VIP/helodermin- and prostaglandin E1-stimulated adenylate cyclase activities in desensitized SUP-T1 human lymphoblasts. AB - VIP/helodermin receptors and PGE1 receptors coupled to adenylate cyclase underwent rapid homologous desensitization and/or down regulation in the human lymphoma SUP-T1 cell line: helodermin- and PGE1-stimulated adenylate cyclase activities in membranes decreased by 75% and 80%, respectively, after a 16-hr incubation of cells with 30 nM VIP or 0.1 microM PGE1. The adenylate cyclase response to helodermin doubled within 120 min of incubation with fresh medium, this part of the resensitization process being not significantly reduced by cycloheximide. The second slower phase of recovery attained 80% of control values after 8 hr and was significantly affected by cycloheximide added at time 0. These data were corroborated by our observations on [125I]helodermin binding to intact cells. In the case of functional PGE1 receptors, sixty percent of the adenylate cyclase response reappeared within 30-60 min, with the second phase of recovery leading, after 2-3 hr to 80-85% of control values of PGE1-stimulated enzyme activity. This resensitization process to PGE1 was, as a whole, cycloheximide sensitive. PMID- 2558362 TI - Decreased adenylate cyclase activation by helodermin and PGE1 in the lectin resistant variant Wa4 of the mouse melanoma cell line B16. AB - We examined the cultured mouse melanoma cell line B16 (clone F1) and its wheat germ agglutinin-resistant variant Wa4 that suffers from abnormal protein glycosylation (a high fucose:sialic acid ratio in glycoproteins). In both cell lines the adenylate cyclase system was endowed with a functional guanine nucleotide binding protein Gs and was efficiently coupled to alpha-MSH receptors. In the B16 cell line F1 studied we also observed an efficient stimulation of adenylate cyclase activity by helodermin, VIP and the VIP analogue [acetyl His1]VIP, and also by PGE1. In membranes from the lectin-resistant variant Wa4, the stimulations by VIP-like peptides and by PGE1 were reduced by 60% and 50%, respectively, while the stimulation by alpha-MSH remained normal. As other components of the adenylate cyclase system (Gs site, catalytical unit) appeared unchanged in the Wa4 variant, we conclude that impaired glycosylation essentially affected the number of both VIP-like peptide receptors and PGE1 receptors. PMID- 2558363 TI - Adrenocorticotropic hormone fragment ACTH/MSH(4-10 can act as a discriminative stimulus in rats. AB - Five of eight male albino rats learned to discriminate the effects of ACTH/MSH(4 10) (100 micrograms/kg, SC) from those of saline vehicle in a food-reinforced T maze task. Generalization tests showed that the ACTH/MSH(4-10) cue was dose related with an ED50 of approximately 68.0 micrograms/kg. These findings indicate that ACTH/MSH(4-10) has interoceptive stimulus properties discriminable to some animals, which might provide a useful model for studying the pharmacology of this neuroactive peptide. Moreover, since ACTH/MSH(4-10) induces effects similar to those of ACTH/MSH in the central nervous system but not hormonally, these data suggest that endogenous ACTH or MSH may also have interoceptive stimulus effects unrelated to peripheral endocrine activity. PMID- 2558364 TI - Autoradiographic distribution of 125I-VIP binding in the rat adrenal cortex. AB - Using in vitro autoradiography, 125I-VIP binding was found to be concentrated in the capsule and glomerulosa of the rat adrenal cortex. The densest receptor distribution was coincident with the distribution of VIP nerve fibers that arborize extensively in the capsule and glomerulosa. The specificity of this binding was demonstrated using unlabelled VIP, ACTH and angiotensin II. The presence and distribution of 125I-VIP binding sites provides the link between the previously found VIP nerves and the steroidogenic effect of exogenous VIP, thereby substantiating the physiological role of VIP-containing autonomic nerves in the regulation of adrenocortical cell function. PMID- 2558365 TI - Effect of neurokinin A on human temporal muscle blood flow. AB - The effect of neurokinin A on human temporal muscle blood flow was compared to saline when injected into the muscle in six normal subjects. The 133-Xenon washout technique was used and the test solutions administered in a double-blind, cross-over manner. Neurokinin A (0.02 ml, 10(-5)M) caused a blood flow increase of 193%, while saline caused an increase of 23%. The difference between neurokinin A and saline was significant (p less than 0.05). It is suggested that a possible pathophysiological role of neurokinin A in migraine must involve modulation of vascular response as well as of primary nociception. PMID- 2558366 TI - Dissociation of the effects of neuropeptide Y on feeding and memory: evidence for pre- and postsynaptic mediation. AB - In mice not deprived of food, centrally administered neuropeptide Y (NPY) increases feeding and improves retention. In this study, we examined the effect of C-terminal NPY fragments on feeding and on memory retention. Mice were trained to avoid footshock in a T-maze. After training NPY, NPY fragments (20-36 and 26 36) or saline were administered intracerebroventricularly. Food consumption was measured during the first hour after training and memory retention was measured one week after training. NPY elicited a 544% increase in feeding compared to the saline control. Neither NPY fragment significantly increased feeding. Both NPY and NPY(20-36) improved retention compared to the saline-treated group. NPY(26 36) did not improve retention. NPY administered to well-trained mice results in amnesia. As a further test of the differential effect of NPY on memory processing and eating, we determined in well-trained mice whether administration of NPY and NPY(20-36) resulted in amnesia. Both NPY and NPY(20-36) resulted in amnesia, but only NPY stimulated feeding. These results are compatible with NPY effects on feeding being mediated through postsynaptic (Y1)NPY receptors and effects on memory retention being mediated through presynaptic (Y2)NPY receptors. PMID- 2558367 TI - Endogenous opioid peptides in the control of food intake in cats. AB - In this report, we investigated the role of exogenous and endogenous enkephalins on food intake in the cat, using, respectively, exogenous [D-Ala2-Met5] enkephalin (DAME) and acetorphan (Ac) in order to inhibit the degradation of endogenous enkephalins. In addition, the selective peripheral antagonist naltrexone methylbromide (NTxMB) and the nonselective antagonist naloxone (Nx) were used in an attempt to discriminate central and peripheral opioid receptors. In 18-hours food-deprived animals, Ac (5 mg/kg IV) increased milk intake during sham feeding (+18%, p less than 0.05), but did not modify it in feeding conditions. Nx (1 mg/kg SC) reduced milk intake in sham-feeding experiments ( 67%, p less than 0.01) more than in milk-feeding conditions (-30%, p less than 0.01). NTxMB (1 mg/kg SC) did not modify milk intake in sham-feeding but decreased it in feeding experiments. In nonfasted animals, Ac did not modify food intake. IV infusion of DAME (50 micrograms/kg) resulted in a reduction of daily food intake (-32%, p less than 0.01). Nx (1 mg/kg SC) decreased the earlier 30 min intake followed by reduction of daily intake (-30%, p less than 0.01). NTxMB (1 and 4 mg/kg SC) increased the 30-min intake dose dependently, without significant change in daily intake. In conclusion, Ac increases food intake in sham-feeding conditions, suggesting that endogenous enkephalins are likely to be involved in the stimulation of food intake. The effects of Nx and NTxMB furthermore suggest both a central activation, and a peripheral inhibition of food intake by opiates when food is allowed to proceed normally through the digestive tract. PMID- 2558368 TI - Induction of V2 receptors in renal medulla of homozygous Brattleboro rats by arginine vasopressin. AB - Homozygous Brattleboro rats display pronounced diabetes insipidus and when treated continuously with arginine vasopressin (AVP) acquire the ability to produce concentrated urine. In this study, the effects of continual AVP replacement on the pharmacological properties of the renal medullary V2 receptor and coupling to adenylate cyclase were examined. Osmotic minipumps that delivered AVP at four different rates were implanted into male homozygous Brattleboro rats. At the end of the 14 day treatment period, urine osmolalities were 280 +/- 24, 474 +/- 105, 1777 +/- 304 and 2202 +/- 175 mOsm/kg H2O for the 0, 31.25, 62.5 and 125 ng/hr treatment groups, respectively. Plasma AVP levels were below the level of detection for the 0 and 31.25 ng/hr treatment groups, and were 2.5 +/- 0.5 and 6.5 +/- 1.8 pg/ml for the 62.5 and 125 ng/hr treatment groups. Saturation experiments using [3H] AVP and renal medullary membranes revealed binding site concentrations of 57 +/- 9, 84 +/- 23, 164 +/- 17 and 150 +/- 18 fmol/mg protein for the 0, 31.25, 62.5 and 125 ng/hr treatment groups, respectively. AVP stimulated cyclic AMP accumulation was enhanced in renal medullary membranes prepared from the 62.5 and 125 ng/hr treatment groups when compared to that in the 0 and 31.25 ng/hr treatment groups. From these results, it appears that circulating AVP is necessary for expression of functional V2 receptors in the homozygous Brattleboro rat renal medulla. PMID- 2558369 TI - Evidence that vascular actions of PHI are mediated by a VIP-preferring receptor. AB - Vasoactive intestinal peptide (VIP) and peptide histidine isoleucine (PHI) are homologous neuropeptides which share vasodilatory properties. This paper addresses the question of whether PHI exerts its vascular action via a receptor distinct from that for VIP. Radioligand binding experiments were done using [Tyr(125I)10]VIP, [Tyr(125I)22]porcine PHI, [Tyr(125I)10]rat PHI and arterial preparations from rat, bovine and porcine species. The radioiodination of rat PHI by the lactoperoxidase-glucose oxidase method and analysis of the structure of the major radiolabeled derivatives were described. All the receptor binding experiments identified a VIP-preferring receptor irrespective of which radioligand or arterial preparation was utilized. VIP and PHI peptides demonstrated cross-desensitization in studies of relaxation of porcine coronary arterial strips in vitro. The present results favor the conclusion that the vascular actions of the PHI peptides are best explained by binding to a VIP preferring receptor. PMID- 2558370 TI - Chemotherapy in lung cancer. PMID- 2558372 TI - Interactions of vitamins A, D3, E, and K in the diet of broiler chicks. AB - A total of 3,888 broiler chicks (Vantress x Arbor Acre) were used in a study involving 81 dietary treatments to determine the interactions among vitamins A, D3, E, and K in broiler chicks. Three levels of each fat-soluble vitamin representing deficient, optimum, and excessive amounts were included. Significant observations were: effect of vitamin A levels on feed efficiency (P less than .01), plasma vitamin A (P less than .01), and plasma vitamin E (P less than .01); effect of vitamin D levels on body weight gain (P less than .01) and mortality (P less than .05); effect of vitamin E levels on plasma vitamin A (P less than .01); effect of vitamin A x vitamin D interaction on body weight gain (P less than .02) and plasma vitamin E (P less than .05); effect of vitamin A x vitamin E interaction on mortality (P less than .01), plasma vitamin A (P less than .03), and plasma vitamin E (P less than .01); effect of vitamin D x vitamin K interaction on feed efficiency (P less than .05); effect of vitamin A x vitamin D x vitamin E interaction on plasma vitamin E concentration (P less than .01); effect of vitamin A x vitamin E x vitamin K interaction on mortality (P less than .05). The results of this study suggest that higher supplemental levels of vitamins D and K would improve performance of poultry occasionally being fed high supplemental levels of vitamins A and E. PMID- 2558371 TI - Recent advances in receptor research. PMID- 2558373 TI - Effect of vitamin C, environmental temperature, chlortetracycline, and vitamin D3 on the development of tibial dyschondroplasia in chickens. AB - Seven experiments were conducted to test the influence of dietary supplementary ascorbic acid on the development of tibial dyschondroplasia in broiler chickens. Ascorbic acid supplementation significantly reduced the incidence and number of birds with a large mass of cartilage in the tibia in the first experiment but not in the two subsequent experiments. Because environmental temperature, microbial infection, and vitamin D3 status had been reported in the literature to influence ascorbic acid metabolism in the chicken, experiments were conducted to see if these variables could influence supplemental ascorbic acid effects on development of tibial dyschondroplasia. Results of the experiments indicated that none of these factors influenced the effect of ascorbic acid on the development of tibial dyschondroplasia. The presence of vitamin D3 in the diet significantly influences the incidence of this disorder. PMID- 2558374 TI - Analytical method for the quantification of 2',3'-didehydro-3'-deoxythymidine, a new anti-human immunodeficiency virus (HIV) agent, by high-performance liquid chromatography (HPLC) and ultraviolet (UV) detection in rat and monkey plasma. PMID- 2558375 TI - Variability in contents of thiamine and riboflavin in brown rice, crude oil in brown rice and bran-polish, and silicon in hull of IR rices. AB - Among 30 IR varieties, variety had greater effect on thiamine content of brown rice than season. Both season and variety affected riboflavin content. Purple- and red-pericarped Philippines rices had higher riboflavin and crude protein contents than IR rices. Crude oil contents of brown rice and bran-polish of 19 IR rices were significantly affected by variety, but only brown rice oil content was significantly affected by season. Bran-polish of waxy rice IR29 had the highest oil content. Silicon content of hull of 16 IR rices showed significant variation due to variety and season and tended to be higher in the dry season crop. Variety rankings in nutrient composition were not consistent for the two crops. The levels of nutrients of IR rices were within the range of values reported for other rices. PMID- 2558376 TI - Characterization of 1,25-dihydroxyvitamin D3 receptors and in vivo targeting of [3H]-1,25(OH)2D3 in the sheep placenta. AB - We sought to detect the presence of receptors for 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] in placental tissues of five late gestational pregnant sheep and to quantitate their biochemical properties and abundance. Cytosol prepared from cotyledonary tissue was found to contain two [3H]-1,25(OH)2D3 binding macromolecules that sedimented at 3.2 S and 4.1 S, respectively, on linear (4-20 per cent) hypertonic sucrose gradients. The 4.1 S component cosedimented with serum that had been prelabelled with [3H]-25-hydroxyvitamin D3 (25-OHD3) and was present in cytosols despite extensive washing of the tissue prior to homogenization. Concurrent incubation of the cytosol with [3H]-1,25(OH)2D3 and a tenfold molar excess of radioinert 25-OHD3 resulted in complete resolution of the 3.2 S macromolecule and disappearance of the 4.1 S binding component. The binding of [3H]-1,25(OH)2D3 to the 3.2 S component was completely abolished by coincubation with a 100-fold molar excess of radioinert 1,25(OH)2D3 and was replaced by a well resolved peak in the 4.1 S region. Scatchard analysis of cytosol binding to [3H]-1,25(OH)2D3 in the presence of a tenfold molar excess of radioinert 25OHD3 revealed a single class of non-interacting saturable binding site in the cotyledon and the endometrium of high affinity and low capacity. The mean +/- s.e. of the dissociation constant of the cotyledonary receptor of 0.21 +/- 0.06 nM was not different from that of 0.16 +/- 0.03 nM for the endometrial receptor. However, the abundance of the cotyledonary receptor was fourfold higher than that in the endometrium (110 +/- 20 versus 28 +/- 7 fmol/mg protein). Since it is not possible to completely separate endometrial tissue from cotyledonary tissue, the low abundance of receptor in endometrial cytosols may merely represent contamination of endometrial tissue with cotyledonary tissue. Further analysis of the [3H]-1,25(OH)2D3 occupied receptor in cotyledonary cytosols showed that it bound to DNA cellulose and was eluted with 0.16 M KCl. This in vitro binding of [3H]-1,25(OH)2D3 to DNA was confirmed in vivo by the finding of preferential nuclear targetting of [3H]-1,25(OH)2D3 (56 per cent of total cellular activity), 4 h after fetal intravenous administration of [3H] 1,25(OH)2D3 to five chronically catheterized fetal sheep. Total placental uptake of [3H]-1,25(OH)2D3 at this time amounted to 3.7 +/- 0.9 per cent of the injected dose. Preliminary analysis of ovine placental cytosols revealed a calcium binding protein of similar molecular weight to that found in the ovine intestine and in the intestine and placenta of rodents.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2558377 TI - Structural analysis of antiviral agents that interact with the capsid of human rhinoviruses. AB - X-Ray diffraction data have been obtained for nine related antiviral agents ("WIN compounds") while bound to human rhinovirus 14 (HRV14). These compounds can inhibit both viral attachment to host cells and uncoating. To calculate interpretable electron density maps it was necessary to account for (1) the low (approximately 60%) occupancies of these compounds in the crystal, (2) the large (up to 7.9 A) conformational changes induced at the attachment site, and (3) the incomplete diffraction data. Application of a density difference map technique, which exploits the 20-fold noncrystallographic redundancy in HRV14, resulted in clear images of the HRV14:WIN complexes. A real-space refinement procedure was used to fit atomic models to these maps. The binding site of WIN compounds in HRV14 is a hydrophobic pocket composed mainly from residues that form the beta barrel of VP1. Among rhinoviruses, the residues associated with the binding pocket are far more conserved than external residues and are mostly contained within regular secondary structural elements. Molecular dynamics simulations of three HRV14:WIN complexes suggest that portions of the WIN compounds and viral protein near the entrance of the binding pocket are more flexible than portions deeper within the beta-barrel. PMID- 2558378 TI - Molecular dynamics simulations of ribonuclease T1: comparison of the free enzyme and the 2' GMP-enzyme complex. AB - Molecular dynamics simulations were performed on free RNase T1 and the 2'GMP RNase T1 complex in vacuum and with water in the active site along with crystallographically identified waters, allowing analysis of both active site and overall structural and dynamics changes due to the presence of 2'GMP. Differences in the active site include a closing in the presence of 2'GMP, which is accompanied by a decrease in mobility of active site residues. The functional relevance of the active site fluctuations is discussed. 2'GMP alters the motion of Tyr-45, suggesting a role for that residue in providing a hydrophobic environment for the protein-nucleic acid interactions responsible for the specificity of RNase T1. The presence of 2'GMP causes a structural change of the C-terminus of the alpha-helix, indicating the transmission of structural changes from the active site through the protein matrix. Overall fluctuations of both the free and 2'GMP enzyme forms are in good agreement with X-ray temperature factors. The motion of Trp-59 is influenced by 2'GMP, indicating differences in enzyme dynamics away from the active site, with the calculated changes following those previously seen in time-resolved fluorescence experiments. PMID- 2558380 TI - Assignment to mouse chromosomes of candidate genes for the rd mutation. PMID- 2558379 TI - Computational and site-specific mutagenesis analyses of the asymmetric charge distribution on calmodulin. AB - Calmodulin's calculated electrostatic potential surface is asymmetrically distributed about the molecule. Concentrations of uncompensated negative charge are localized near certain alpha-helices and calcium-binding loops. Further calculations suggest that these charge features of calmodulin can be selectively perturbed by changing clusters of phylogenetically conserved acidic amino acids in helices to lysines. When these cluster charge reversals are actually produced by using cassette-based site-specific mutagenesis of residues 82-84 or 118-120, the resulting proteins differ in their interaction with two distinct calmodulin dependent protein kinases, myosin light chain kinase and calmodulin-dependent protein kinase II. Each calmodulin mutant can be purified to apparent chemical homogeneity by an identical purification protocol that is based on conservation of its overall properties, including calcium binding. Although cluster charge reversals result in localized perturbations of the computed negative surface, single amino acid changes would not be expected to alter significantly the distribution of the negative surface because of the relatively high density of uncompensated negative charge in the region around residues 82-84 and 118-120. However, this does not preclude the possibility of single amino acid charge perturbations having a functional effect on the more intimate, catalytically active complex. The electrostatic surface of calmodulin described in this report may be a feature that would be altered only by cluster charge reversal mutations. Overall, the results suggest that the charge properties of calmodulin are one of several properties that are important for the efficient assembly of calmodulin protein kinase signal transduction complexes in eukaryotic cells. PMID- 2558381 TI - Failed assembly of phosphodiesterase complex in developing photoreceptors of rd mice. PMID- 2558383 TI - Localization of 5'-nucleotidase activity in RCS rat retinas. PMID- 2558382 TI - Functional and biochemical abnormalities in the retinas of mice heterozygous for the rd gene. PMID- 2558385 TI - Intense-light mediated changes in rat rod outer segment lipids and proteins. PMID- 2558386 TI - Photoreceptor cell rescue in the RCS rat by RPE transplantation: a therapeutic approach in a model of inherited retinal dystrophy. PMID- 2558384 TI - Analysis of normal and rcdl Irish setter retinal proteins. AB - In both of the early onset systems, the rd mouse and the rcdl Irish Setter, early elevation of cyclic GMP may be the ultimate cause of accelerated photoreceptor degeneration. This would be consistent with the data utilizing in vitro systems in which retinal samples, in culture, undergo degeneration in response to constant exposure to high levels of this nucleotide. However, the ultimate cause of the elevated cyclic GMP in the rd mouse or in the rcdl Irish Setter still remains a mystery. It appears that all of the necessary proteins of the visual cascade are produced, although they are lost at different rates. The phosphodiesterase appears to be reduced faster than other proteins. This may, in turn, account for the elevation in cyclic GMP levels. The cause of this enhanced disappearance could reside in the phosphodiesterase protein itself, or in other more distal components. The alteration in rhodopsin reaction to the specific rhod 4 antisera suggests that this protein is not properly oriented in the disc membrane. Although this may or may not alter the visual cascade, it does suggest that these membranes are not identical to those of the normal dog retina. Future studies should focus on the individual functional activities of each component, on their structures, and on their proper assembly within the disc. PMID- 2558388 TI - Human monocytes convert leukotriene B4 to two dihydro-leukotriene B4-metabolites. AB - Human monocytes metabolize LTB4 by an additional pathway different from omega oxidation. Reverse-phase high performance liquid chromatography showed four metabolites: 20-COOH-LTB4, 20-OH-LTB4 and two metabolites less polar than LTB4 with an UV maximum at 232 nm. Gas-chromatography mass-spectrometry showed nearly identical mass spectra for both metabolites. The main mass fragments of the two metabolites were increased by two mass units compared to LTB4. Our findings suggest that LTB4 had been reduced to a known and a new dihydro-metabolite of LTB4. Both metabolites together amounted to 85% of total metabolites. The remaining 15% were omega-oxidation products. Thus, the major pathway of LTB4 metabolism by human monocytes is reduction to dihydro-LTB4. PMID- 2558387 TI - Evidence of direct positive inotropic and chronotropic actions by PGE1 not mediated by cyclic AMP in conscious sheep. AB - The underlying mechanism for the cardiac responses to PGE1 has not yet been fully elucidated. In order to investigate a possible role for cyclic AMP in the positive inotropic and chronotropic actions of prostaglandin E1 (PGE1) in conscious sheep, theophylline-ethylenediamine was used to inhibit phosphodiesterase activity. Any significant potentiation or the lack of potentiation of the measured cardiac response to PGE1 was then used as a criterion to establish whether the cardiac actions of PGE1 were produced by an alteration in the intracellular levels of cyclic AMP. The results suggest that PGE1 produced positive inotropic and chronotropic actions in conscious sheep, which is neither caused by the autonomic nervous system (baroreflexes) nor by changes in intracellular cyclic AMP levels. Further research seems warranted to establish whether a relationship exists between Ca2+ and the contractile response of PGE1. Such a relationship could then possibly explain the positive inotropic action of PGE1 in conscious sheep. PMID- 2558389 TI - Prostaglandin catabolism in fetal and maternal tissues--a study of 15 hydroxyprostaglandin dehydrogenase and delta 13 reductase with specific assay methods. AB - Recent studies have demonstrated that extraductal tissues such as lung are important sources of prostaglandin E2 which maintains the patency of ductus arteriosus in fetuses and prematurely-born infants. Also, organs such as lung are known to be active in the catabolism of PGE2. Earlier studies of enzymes involved in the catabolism of PGE2 such as 15-hydroxyprostaglandin dehydrogenase (15-PGDH) and delta 13 reductase all used non-specific methods. In the present report, we studied 15-PGDH in fetal and maternal rat lung, kidney, and fetal lamb lung, kidney and ductus arteriosus with the use of a specific substrate (15-S)-[15(3)H PGE2]. In addition, we measured the activity of delta 13 reductase in these tissues by measuring the conversion of [1-14C]-15-keto PGE2 to [1-14C]-15-keto 13,14-dihydro PGE2. The results from these studies demonstrated that in fetal rat lung and kidney, 15-PGDH activities increased rapidly while delta 13 reductase remained unchanged during late gestation. Ductus arteriosus possessed little 15 PGDH activities. These results strongly suggest that extraductal regulation of PGE2 metabolism is important in determining ductal caliber in fetuses and prematurely delivered neonates. PMID- 2558390 TI - Kinetic resolution of racemic leukotriene A4 by mammalian cytosolic epoxide hydrolases. AB - Cytosols of rat and guinea pig liver and of human placenta were screened for their capacity to catalyze the conversion of racemic leukotriene A4 into 5S, 12R dihydroxy-(Z,E,E,Z)-6,8,10,14-eicosatetraenoic acid (leukotriene B4). The epoxide hydrolase activities showed some specificity for the 5S,6S-oxido-(E,E,Z,Z) 7,9,11,14-eicosatetraenoic acid (LTA4) and produced mixtures of leukotriene B4 and its enantiomer containing up to 78-87% of leukotriene B4. PMID- 2558391 TI - Excitatory amino acid receptors in the vertebrate central nervous system. PMID- 2558392 TI - Local anesthetics (benzyl alcohol, lidocaine, procainamide) inhibit aminopyrine accumulation in isolated rat parietal cells. AB - The present studies were designed to examine the effect of local anesthetics (benzyl alcohol, lidocaine, and procainamide) on the secretory response of parietal cells to histamine, dbcAMP, and carbachol. Studies were performed in vitro using isolated cells from rat stomachs, and acid production was determined by 14C-aminopyrine accumulation. In addition, the (H(+)-K+)-ATPase activity of microsomal vesicles isolated from parietal cells was determined. Lower concentrations of the drugs studied increased the basal aminopyrine accumulation and potentiated the secretory response of parietal cells to histamine and dbcAMP. At higher concentrations local anesthetics progressively inhibited both the basal 14C-aminopyrine accumulation and that stimulated by histamine, dbcAMP or carbachol. While a low concentration of local anesthetics increased gastric microsomal (H(+)-K+)-ATPase activity, higher concentrations inhibited enzyme activity to about 80% of those activities found in resting parietal cells. We conclude that increased aminopyrine accumulation may reflect the activation of membrane-bound enzyme(s) involved in the cAMP-dependent signal transduction pathway mediating acid secretion by parietal cells. In turn, it is possible that the inhibition of aminopyrine accumulation by local anesthetics at higher concentrations can relate to two different mechanisms: (1) the nonspecific effect of local anesthetics that causes simple proton neutralization (as weak bases), and (2) to a minor extent their inhibitory effect on proton pump activity. PMID- 2558393 TI - [Full inpatient versus partial inpatient psychiatric after-care--a comparative retrospective study]. AB - In a retrospective study 80 inpatients (Sociotherapy, Therapeutic Community) were compared to 160 outpatients (80 patient Day Clinic, 80 patients Night Clinic). With a mean treatment duration of 4-5 1/2 month a significant decrease was found for each group in the frequency and duration of further hospital admissions after end of therapy. Day and Night Clinic patients tending to a better outcome than the inpatients. Advantages of the Day and Night Clinic are seen e.g. in a higher degree of acceptance by patients and family doctors, disadvantages in the increased rate of suicide found in our sample. PMID- 2558394 TI - [Intracavitary afterloading-therapy of cervicofacial tumors--results and experiences to date]. AB - Since 1984 we have used the intracavitary afterloading brachytherapy also in therapy of cervicofacial tumors. We report on 21 patients, 12 of them with therapy of recurrences and 9 with a primary therapy. The patients represent an inhomogeneous selected group. With recurrences also long tumor-free survival periods (till now 44, 52, 53 months) can be attained by means of contact therapy alone or in combination with percutaneous irradiation. Also after primary intensive irradiation curative radiation doses are applicable again as therapy of recurrences. Procedure can likewise be applied in primary therapy of cervicofacial tumors as a boost-irradiation in percutaneous radiotherapy. It should be reserved to limited tumor situations in compliance with irradiation of small volumes. PMID- 2558395 TI - [Series CT of hormone-active pancreatic tumors]. AB - Localisation of hormonally active pancreatic tumours with serial CT has gained practical significance, although these tumours are rare. Its sensitivity was approximately 80%. The methodologic details and problems of image analysis and interpretation are discussed. The analysis of pathologic findings, normal vessel topography and artefacts is shown to improve the diagnosis of hormonally active pancreatic tumours. PMID- 2558396 TI - Opioid peptides and receptors. Localization, interactions and relationships to other molecules in the rodent brain, especially the hippocampal formation. PMID- 2558397 TI - [Molecular assembly of Na+, K(+)-ATPase and its dynamic aspects--determination of membrane protein molecular weight by low-angle laser light-scattering photometry]. PMID- 2558398 TI - [Primary tumor of the liver. Description of a case with atypical clinico radiological onset]. PMID- 2558399 TI - Nedocromil sodium workshop. PMID- 2558401 TI - Malignant fibrous histiocytoma of the anterior mediastinum: a rare case with 19 years survival. PMID- 2558400 TI - Nedocromil sodium: an overview. PMID- 2558402 TI - [Lymphomatous hepatic infiltration]. PMID- 2558403 TI - [Sepsis caused by Clostridium in patients with neoplasms]. PMID- 2558404 TI - [Meta-analysis on randomized trials comparing the results of low-molecular weight heparins to those of fractioned heparins in the prevention of deep venous thrombosis]. AB - The main reason for using low molecular weight heparin (LMWH) is its capacity to keep the antithrombotic effect of standard non-fractioned heparin (NFH) while reducing its hemorrhagic power. Various studies comparing LMWH to NFH gave contradictory results, so we carried out a meta-analysis of randomized trials comparing these two treatments in prevention of deep-vein thrombosis (DVT) (and in prevention of deep-vein thrombosis or pulmonary embolism). From a selection of ten trials we could not show any significant difference in the DVT rates, as well as for the whole of surgical indications as for the subgroups of abdominal and orthopedic surgery. The weighted global difference between the DVT rates is so low that, in spite of the use of meta-analysis, the power is clearly insufficient. The hemorrhagic risk is not statistically reduced as well for the whole of surgical indications as for the subgroups. PMID- 2558405 TI - Acute and chronic effects of eicosapentaenoic acid (EPA) on the cardiovascular system. AB - Several studies have shown that the development of high blood pressure in spontaneously hypertensive rats (SHR) can be attenuated by feeding them modified fat diets. In the present study, eight weeks after treatment with eicosapentaenoic acid (EPA; 100 mg/kg/day s.c.) SHR had lower systolic blood pressure (BP); (tail-cuff plethysmography) compared to saline-injected SHR: 180.0 +/- 2 vs. 204.0 +/- 1 mmHg, respectively, (p less than 0.001). There was no significant difference in the BP of EPA and saline-treated WKY (Wistar-Kyoto) rats. Heart rate (HR) decreased with age in both the SHR and WKY rats and no significant effect of EPA was observed in WKY rats; the decrease in heart rate in the SHR group was significantly diminished. EPA did not significantly alter growth rate of SHR and WKY rats. However, aged-matched WKY rats weighed more than the SHR. The pressor responses to norepinephrine in doses of 0.3 and 3.0 microgram/kg, i.v., as well as plasma NE, DOPA and MHPG, were also not significantly affected by EPA-supplementation in both SHR and WKY rats. Data from the present study support the view that EPA might be an effective treatment of hypertension that develops via mechanisms unrelated to sympathetic activity or vascular reactivity to adrenergic neurotransmitters. PMID- 2558406 TI - Evaluation of zinc complexes on the replication of rhinovirus 2 in vitro. AB - The effect of zinc salts and complexes were evaluated on the replication of rhinovirus 2 in vitro. Zinc chloride inhibited the replication of rhinovirus 2 at concentrations between 3 and 12 micrograms/ml. Influenza virus was not affected. A number of zinc complexes were tested and compared to zinc chloride. The results indicated that the activity and toxicity of all zinc complexes in the rhinovirus cytopathogenic effect (CPE) assay were directly related to the amount of unbound zinc available. PMID- 2558407 TI - [Psychosine: a "toxin" produced in the brain--its mechanism of action]. AB - Psychosine (galactosylsphingosine) is enzymatically synthesized from UDP galactose and sphingosine and degraded by galactosylceramidase. Galactosylceramidase is genetically deficient in Krabbe disease (globoid cell leukodystrophy) and psychosine accumulates in the brain of humans, dogs or mice affected by the disease. Psychosine has a very potent inhibitory effect on cytochrome c oxidase (COX) in mitochondria. When COX is purified, psychosine does not suppress the enzyme activity. However, a clear inhibitory effect is seen when the enzyme is "reconstituted" with sonicated phosphatidylcholine, suggesting that the inhibition of the enzymatic activity is caused by perturbation of the environment of COX in the mitochondrial membrane. Studies using analogues suggest that the free amino group in the sphingosine moiety plays an important role in exerting the effects of psychosine. The effects of psychosine are not only potent but fast and reversible. It is noteworthy that a powerful inhibitor of cellular respiration is synthesized in mammals' brain, an organ vulnerable to hypoxia. PMID- 2558408 TI - Augmentation of lymphocyte and macrophage proliferation by caprine arthritis encephalitis virus contributes to the development of progressive arthritis. AB - Infection by the lentivirus, caprine arthritis-encephalitis virus (CAEV), may lead to an intermittent arthritis due to the presence of lymphocytes and macrophages in synovial membranes. We have observed that the presence of CAEV increases the division of lymphocytes and macrophages. In association, antigen induced arthritis is more severe in goats concurrently infected by CAEV than in noninfected goats. From these observations we propose that infection by this lentivirus increases reactivity of lymphocytes and macrophages to immune and nonimmune stimuli, leading to the increased likelihood of progressive arthritis in infected animals. PMID- 2558409 TI - Pathogenesis of lentivirus-induced arthritis. A review. AB - While it has been known for some years that there is an association between lentiviruses and slowly progressive joint diseases in ruminants, the realization that the human immunodeficiency virus, the cause of AIDS, is a lentivirus has made this group of virus the focus of a considerable research effort. The manifestations of lentivirus infection in animals are discussed and reference is made to the possibility of using them as models for human rheumatoid arthritis and for AIDS. PMID- 2558411 TI - Activity of some lysosomal enzymes and postnatal ontogenetic development of epileptic reactivity in rabbit brain. AB - The beta-glycerophosphatase (GP), p-nitrophenyl phosphatase (PNFP) and beta glucuronidase (BG) activities in four neocortical rabbit areas have been determined in six successive electrocorticographic stages of postnatal ontogenetic development of epileptic reactivity. The lysosomal enzyme activity decreases gradually from rabbits aged 1 day to adult ones and is similar within the four neocortical areas. The activity of the three lysosomal enzymes is not parallel. Statistically significant decreases in beta-glycerophosphatase activity are present in rabbits at the age of 30 days versus adult age in all the areas investigated while beta-glucuronidase is higher at the same developmental stage only in beta retrosplenial (III) and motor (II) areas. Correlation of the electrophysiological and biochemical data cannot provide explanation for the particular phenomena of the epileptogenic reactivity in rabbit cerebral neocortex. PMID- 2558412 TI - [Calcium ions, calcium antagonists and ischemic arrhythmia]. PMID- 2558410 TI - Differential immunological response of patients with rheumatoid arthritis towards two different Epstein-Barr virus strains: inhibition of interleukin-1 release by the B95-8, but not the P3HR-1 virus strain. AB - An abnormal immune response towards the Epstein-Barr virus (EBV) has been documented in patients with rheumatoid arthritis (RA). To investigate whether these findings are due to the transformation event caused by EBV, RA blood mononuclear cells and monocyte-enriched preparations were incubated with two different EBV strains: the transforming virus secreted by the cell line B95-8 and the virus released by the P3HR-1 cell line that is not able to transform due to a small deletion in the U2 region of the virus genome. Immunological response was determined by the production of interleukin-1 (IL-1) and tumor necrosis factor (TNF) using ELISA and bioassay systems. There was a striking difference in cytokine measurements with a strong inhibition of IL-1 and TNF production after incubation with the B95-8 virus, but not the P3HR-1 virus. These data indicate that the disturbed reaction of the immune system towards EBV is either dependent on the full transformation of B cells in RA patients or alternatively due to the secretion of a cytokine inhibitor by the B95-8 cell line. PMID- 2558413 TI - [Biochemistry and physiopathology of thrombomodulin]. PMID- 2558416 TI - The selenium method for treatment of lakes for elevated levels of mercury in fish. AB - Selenium in the form of sodium selenite (Na2SeO3) incorporated in a rubber matrix was introduced into a Swedish lake which is black-listed for fishing because of high mercury levels in fish tissues. The lake is not acidified. The level of selenium in the lake was raised from 0.4 to approximately 3-5 micrograms Se l-1, and sustained at this level for a period of greater than 3 years. The mercury content of pike (Esox lucius) was found to have fallen markedly after the lake had been treated for only 1 year. The mercury content was halved over the whole weight range and stayed below the black-list limit during the treatment period. After the first year the mercury content of perch (Perca fluviatilis) muscle had also decreased. The reduction for those between 8 and 11 cm long (3-6 years old) was found to be 77%; for those between 12 and 16 cm long (7-9 years old), 65%; and for those between 17 and 25 cm long (greater than 9 years), 77%. After 2 years, the total reduction was 85% for perch 12-16 cm long and 75% for those 17 25 cm long. By the end of the third year, the mercury content had decreased by 84% in those that measured between 17 and 25 cm. The selenium treatment also resulted in a very marked decrease in the mercury content of the muscle of roach (Leuciscus rutilus). The decrease, from 0.5-1.0 to 0.06-0.08 mg Hg/kg muscle, was independent of size. The treatment of lakes with selenium has been found to be one of the few known methods of lowering the mercury content of fish quickly and easily. PMID- 2558415 TI - [One type of excitatory amino acid receptor]. PMID- 2558414 TI - [Electrophysiologic properties of pancreatic beta-cells]. PMID- 2558417 TI - Cholangiocarcinoma masquerading as liver abscess. AB - A 60-year-old man from Eastern Thailand was admitted to hospital because of right upper quadrant abdominal pain and fever. Ultrasonographic examination revealed two cavitary lesions in the right lobe of the liver. Needle aspiration obtained 110 ml of anchovy sauce-like pus which showed no bacteria on gram stain and routine culture. Serological test for E. histolytica antibody was negative. Initially, the patient responded well to metronidazole. Two weeks later, the symptoms recurred and sonography revealed one large cavitary lesion with three adjacent locules in the right lobe of the liver. Repeated needle aspiration again showed anchovy sauce-like pus which grew Enterobacter agglomerans. O. viverrini ova were detected in the stool. Laparotomy revealed histologically proven cholangiocarcinoma. This report indicates that O. viverrini infection associated with CCC can masquerade as liver abscess. PMID- 2558418 TI - Phyllode tumours of the breast. A review of 21 cases. AB - Twenty-one cases of phyllode tumours of the breast (9 malignant, 4 borderline and 8 benign) were reviewed. Three patients with malignant tumours developed metastases, and all died. Eight patients developed local recurrence, 6 in the malignant and borderline groups and 2 in the benign group. The classification into malignant, borderline and benign groups is a reliable guide for use when deciding on the extent of surgery. Malignant and borderline tumours require local mastectomy, while tumour excision suffices for benign disease. The customary pathological separation of malignant phyllode tumours from primary sarcoma of the breast is questioned. PMID- 2558419 TI - Giant fibro-adenomas in black and Indian adolescents. AB - Over a 5-year period, 47 giant fibro-adenomas were managed in the benign breast clinic at King Edward VIII Hospital, Durban. These tumours usually present in the perimenarchal period with gross breast asymmetry; smaller fibro-adenomas are present in 10% of these patients. A hormonal basis for their development is likely. Local excision with preservation of the breast tissue is recommended. PMID- 2558420 TI - Motor effects of two sigma ligands mediated by nigrostriatal dopamine neurons. AB - (+)-Pentazocine, a potent sigma ligand that lacks affinity for PCP receptors, produced dose-dependent contralateral circling behavior following microinjections in the substantia nigra of rats. This effect was attenuated by 6-hydroxydopamine (6-OHDA) lesions of ascending dopamine neurons and enhanced by systemic injections of amphetamine, 6-OHDA lesions also attenuated the circling produced by another selective sigma ligand, 1,3-di-o-tolylguanidine (DTG). These findings suggest that sigma receptors are involved in the neural control of movement and the regulation of the ascending dopamine system. Since all typical antipsychotic drugs tested bind to sigma receptors with Ki values less than 1 microM, these findings further suggest that sigma receptors may mediate some of the motor side effects of antipsychotic drug therapy. PMID- 2558421 TI - Ontogeny of oxytocin receptors in rat forebrain: a quantitative study. AB - The ontogeny of oxytocin receptors in rat forebrain was studied using the selective oxytocin receptor antagonist 125I-d(CH2)5[Tyr(Me)2, Thr4, Tyr-NH29]OVT [( 125I]-OTA). With in vitro receptor autoradiography, binding wa noted on the first postnatal day in dorsal subiculum and thalamus. On postnatal days 5-18, intense labeling was evident in posterior cingulate cortex, dorsal subiculum, lateral septum, and the CA1 subfield of hippocampus. Of these regions only the lateral septum expressed oxytocin receptors in adult brain. Competition studies on coronal sections through posterior cingulate, septum, and dorsal subiculum at P10 demonstrated that transient binding sites in these areas were indeed oxytocin selective (OXY greater than AVP greater tha V1 greater than V2). Result of saturation studies on cingulate membranes from 10-day-old pups agreed favorably with previous reports of the kinetics of [125I]-OTA binding to adult oxytocin receptors (Kd = 0.1 nM in P10 cingulate cortex vs. 0.07 nM for adult ventral subiculum). In contrast to these evanescent developmental sites, oxytocin receptors in the bed nucleus of the stria terminalis and the ventromedial nucleus of the hypothalamus only appeared in adulthood, presumably in response to the surge of gonadal steroids at puberty. PMID- 2558422 TI - [Primary cholangiocellular cancer of the liver with an eosinophilic leukemoid reaction]. PMID- 2558423 TI - The comparison of the pharmacokinetics of a low molecular weight heparin in the newborn and adult pig. AB - Standard heparin (SH) is frequently used in the sick neonate to prevent catheter related thrombosis. SH can cause significant bleeding complications in the adult and its use in the neonate is linked to an increased incidence of intraventricular hemorrhage. Recently available low molecular weight heparins (LMWH) offer potential advantages over SH in the adult by exhibiting a longer half life and decreased bleeding side effects compared to SH. Whether LMWHs would offer similar therapeutic advantages to the sick neonate is unknown. Using the porcine model of neonatal hemostasis we measured the pharmacokinetics of a LMWH (Choay 222) in the pig (ATIII level: 100%), in the piglet (ATIII level:50% of adult) and in the piglet given exogenous ATIII. All pigs were bolused with 125I LMWH (5, 25 or 100 anti-factor Xa units/kg) and blood samples collected for the measurement of 125I-radioactivity, and anti-factor Xa activity. The half life of LMWH, measured as either 125I-radioactivity or as anti-factor Xa activity, was not dose dependent and was similar in pigs and piglets; however, the volume of distribution was greater in the piglet resulting in an increased total clearance compared to the pig. As well, the supplementation of the piglet with exogenous ATIII did not influence the pharmacokinetics of LMWH. The half life of the LMWH in both pigs and piglets was approximately twice as long as previously reported values for SH in the same animal model. Thus the longer half life of LMWH in the piglet, and the similarity of the half life in piglets and pigs suggest that LMWH may have a therapeutic advantage in the newborn over SH. PMID- 2558424 TI - Influence of a low molecular weight heparin on the inhibition of factor Xa and thrombin in hip surgery. PMID- 2558425 TI - [Rotavirus in stools from hospitalized children]. AB - Electron microscopy, ELISA for rotavirus and virus cultivation in cell cultures were used to study the occurrence of rotavirus in 451 fecal samples collected from hospitalized children. Rotavirus was found in samples from 24 of 27 patients admitted with acute diarrhoea. All specimens from randomly selected patients without diarrhoea were negative. Two patients developed nosocomial rotavirus infection. ELISA was positive, but electron microscopy was negative in two specimens obtained from asymptomatic newborns. In this investigation rotavirus was the dominating agent isolated in infectious gastroenteritis. We found no rotavirus excretors without diarrhoea. PMID- 2558426 TI - Age-related reduction in the number of rabbit erythrocyte Na, K-ATPase. AB - The number of Na, K-ATPase units, assessed by [3H]-ouabain binding assay, was significantly less in erythrocytes from 17 mature (30-week) rabbits than in those from 17 young (16-week) ones (0.179 +/- 0.010 vs. 0.263 +/- 0.014 pmol/10(9) cells, p less than 0.01). Consistent with this finding, the erythrocyte sodium concentration was significantly higher in mature rabbits as compared with young animals (17.1 +/- 1.30 vs. 10.8 +/- 0.68 mmol/liter, p less than 0.01). When the assays were repeated in 8 of the young rabbits 8 weeks after the initial determination, both variables were found to be similar to the levels from mature animals. The data suggest that the activity of erythrocyte sodium pump declines with aging or growth of donor rabbits, as a result of a reduction in the number of Na, K-ATPase units. PMID- 2558427 TI - Effect of verapamil on organophosphorus-induced delayed neuropathy in hens. AB - Verapamil, a calcium channel blocker, was administered to adult white leghorn hens to determine if inhibition of calcium entry could alter delayed neuropathy induced by administration of phenyl saligenin phosphate (PSP). Verapamil was given im in doses of 7 mg/kg/day for 4 days beginning 24 hr before administration of PSP (2.5 mg/kg im). Ataxia was less pronounced in hens given PSP plus verapamil than in hens given PSP alone during observations made 8-28 days after PSP administration. Myelinated fiber lesions were less extensive and regeneration more notable in the biventer cervicis nerve in chickens given PSP plus verapamil, with samples obtained both 17 and 28 days after PSP. In the absence of verapamil, rheobase and chronaxie values of strength-duration curves were higher and shorter, respectively, and sensitivity to acetylcholine was increased in biventer cervicis nerve-muscle preparations from hens given PSP. Verapamil did not alter PSP-induced inhibition of neurotoxic esterase, indicating that the mechanism involved in amelioration of these indices of delayed neuropathy was not associated with initial enzyme inhibition caused by this organophosphorus ester. PMID- 2558428 TI - Influence of 2,3-dimercaptosuccinic acid on gastrointestinal lead absorption and whole-body lead retention. AB - 2,3-Dimercaptosuccinic acid (DMSA) is a new orally active heavy metal chelator for the treatment of childhood Pb intoxication on an outpatient basis. The influence of DMSA, as well as other chelating agents, on gastrointestinal 203Pb absorption and whole-body 203Pb retention was examined. Groups of Sprague-Dawley rats (230-260 g) were gavaged with a solution containing approximately 25 mg/kg Pb [as Pb(NO3)2] plus 15 microCi 203Pb. Some groups were then immediately given 0.11 mmol/kg of either DMSA, CaNa2EDTA, D-penicillamine, or BAL by oral gavage, while other groups received the same drugs by ip injection. Control groups received solutions of the drug vehicles po or ip. Whole-body Pb retention and gastrointestinal Pb absorption (whole body retention + urinary Pb excretion) were significantly decreased in rats that received DMSA po. This finding implies that the use of DMSA to treat childhood lead intoxication on an outpatient basis is not associated with a risk for increased Pb absorption. PMID- 2558429 TI - 2,2',4,4',5,5'-hexachlorobiphenyl as a 2,3,7,8-tetrachlorodibenzo-p-dioxin antagonist in C57BL/6J mice. AB - At doses as high as 750 to 1000 mumol/kg, 2,2',4,4',5,5'-hexachlorobiphenyl (HCBP) did not cause fetal cleft palate, suppress the splenic plaque-forming cell response to sheep red blood cells, or induce hepatic microsomal ethoxyresorufin O deethylase (EROD) in C57BL/6J mice. Despite the lack of activity of HCBP in eliciting any of these aryl hydrocarbon (Ah) receptor-mediated responses, competitive binding studies indicated that HCBP competitively displaced 2,3,7,8 [3H]tetrachlorodibenzo-p-dioxin (TCDD) from the murine hepatic cytosolic receptor. Cotreatment of C57BL/6J mice with TCDD (3.7 nmol/kg) and HCBP or 4,4' diiodo-2,2',5,5'-tetrachlorobiphenyl (I2-TCBP) (400 or 1000 mumol/kg) showed that both compounds partially antagonized TCDD-mediated cleft palate and immunotoxicity (i.e., suppression of the splenic plaque-forming cell response to sheep red blood cells), and HCBP antagonized TCDD-mediated hepatic microsomal EROD induction. Thus, HCBP and I2-TCBP, like the commercial polychlorinated biphenyl mixture Aroclor 1254, were partial antagonists of TCDD action in C57BL/6J mice; however, it was also apparent from the results that Aroclor 1254 was the more effective antagonist at lower doses. Using [3H]TCDD, it was also shown that some of the effects of HCBP on TCDD-mediated cleft palate may be due to the decreased levels of TCDD found in the fetal palates after cotreatment with TCDD and HCBP. 4,4'-[125I2]diiodo-2,2',5,5'-tetrachlorobiphenyl ([125I2]TCBP) of high specific activity (3350 Ci/mmol) was synthesized and used to investigate the direct binding of this compound to the murine hepatic Ah receptor or other cytosolic proteins. No direct specific binding was observed between 125I2-TCBP and any cytosolic proteins using a sucrose density gradient assay procedure. These results contrasted with previous studies with Aroclor 1254 that suggested that this mixture acted as a competitive Ah receptor antagonist. PMID- 2558430 TI - The mobilization of intracellular cadmium by butyl and amyl esters of meso-2,3 dimercaptosuccinic acid. AB - The esters of the general structure, [CH(SH)COOR]2, i.e., Di-BDMS, R = CH2CH(CH3)2; Ds-BDMS, R = CH(CH3)CH2CH3; Di-ADMS, R = CH2CH2CH(CH3)2; and D3 ADMS, R = CH(CH2CH3)2 from the reaction of meso-2,3-dimercaptosuccinic acid with isobutyl, sec-butyl, isoamyl, and 3-amyl alcohols, respectively, have been prepared, characterized, and examined as chelating agents for the removal of cadmium from its aged intracellular deposits. All of these compounds depleted cadmium from such deposits and significantly reduced the whole body levels of cadmium. In the case of three (Ds-BDMS, Di-BDMS, and Di-ADMS) of these compounds, the reductions achieved are equal to or greater than that produced by 2,3 dimercapto-1-propanol (BAL) under similar circumstances. None of these compounds caused any redistribution of cadmium to the brain, and two of them (Di-BDMS and Di-ADMS) caused a very much larger reduction in the liver levels of cadmium than BAL. None was as effective as BAL in reducing kidney levels of cadmium. These compounds are not soluble in water and are administered as solutions in peanut oil. A comparison of the behavior of these compounds with others which have been reported to be effective in reducing body burdens of cadmium in chronic cadmium intoxication reveals that they are among the most effective. An analysis of the manner in which mobilizing efficacy changes with structure indicates that higher, purely alkyl analogs are not expected to be superior to these compounds, though other structural variations may be. PMID- 2558431 TI - Chemotherapy-induced premature ovarian failure: mechanisms and prevention. AB - Significant advances have been made in the previously unexplored areas of the mechanisms involved in cyclophosphamide (CTX)-induced ovarian toxicity and the protective effects of luteinizing hormone-releasing hormone (LHRH agonists. The structure and function of granulosa cells and oocytes are affected by the chemotherapeutic agent, CTX. Results of experiments in female rats indicate that LHRH agonists may protect the ovaries from the toxic effects of chemotherapy. The protective effect may be related to the inhibition of ovarian mitotic activity during LHRH agonist administration. This inhibition is much more pronounced in female compared to male rats. This may be related to the observed better gonadal protective effects in females compared to males. Further experiments are underway to determine whether similar protective effects occur in female primates. PMID- 2558432 TI - Effects of hormones and intracellular mediators on differentiated functions of cultured Leydig tumor cells. AB - Cellular regulation by hormones that utilize a myriad of intracellular signaling pathways is recognized to be quite complex. To investigate some of these effects in an established cell line, we tested a panel of hormones and modulators for their effects on cyclic AMP (cAMP) and progesterone production, both alone and in combination with human chorionic gonadotropin (hCG), using the MA-10 cultured Leydig tumor cell line. None significantly affected intracellular levels of cAMP, and only epidermal growth factor (EGF) and 12-O-tetradecanoyl-phorbol-13-acetate (TPA) stimulated progesterone production. While EGF, basic fibroblast growth factor, insulin, insulin-like growth factor-1, and transforming growth factor beta all decreased cAMP production only, TPA decreased hCG-stimulated cAMP and progesterone production. Those factors that stimulated progesterone production also induced a characteristic morphological change ("rounding") of these cells. In addition, EGF, insulin, and TPA, like hCG, elevated mRNA levels of competence oncogenes (c-fos and c-myc), albeit to different extents. These data demonstrate the wide range of hormones to which the cultured Leydig tumor cell will respond, as well as the varying degree of responses observed in the intracellular signaling pathways that we examined. PMID- 2558434 TI - Suitability of the Phast system for discrimination of Entamoeba isolates by isoenzyme isoelectrofocusing. PMID- 2558433 TI - Reversal of drug resistance in Trypanosoma cruzi and Leishmania donovani by verapamil. AB - Following previous studies of verapamil reversal of chloroquine resistance in malaria and multi-drug resistance in cancer cells, the effect of verapamil was investigated on nifurtimox-resistant Trypanosoma cruzi in vitro and antimony resistant Leishmania donovani in vitro and in vivo. Verapamil alone was not active against either parasite, but in combination with nifurtimox it reversed the drug resistance of T. cruzi and in combination with sodium stibogluconate reversed the drug resistance of L. donovani. PMID- 2558436 TI - The E. coli dnaA initiation protein: a protein for all seasons. PMID- 2558435 TI - Cytomegalovirus antibodies in Nigeria. PMID- 2558437 TI - [Klippel-Trenaunay-Weber syndrome. Magnetic resonance imaging diagnosis of medullary involvement]. AB - The Klippel-Trenaunay-Weber (KTW) syndrome is a rare congenital syndrome of unknown etiology consisting of the triad: a large cutaneous naevus, congenital varicosities and hypertrophy of bones and soft tissues. A heterogenous group of vascular malformations may also occur. The case record of acute myelopathy in a patient aged 42 years with recognized KTW syndrome is presented. It is concluded that magnetic resonance imaging is indicated in cases of suspected intramedullary haemorrhage in patients with congenital vascular malformations. PMID- 2558438 TI - Adrenal venous sampling and adrenal biopsy. AB - The current status of adrenal vein sampling and the role of adrenal gland percutaneous biopsy will be reviewed. PMID- 2558439 TI - Necrotic hepatitis of rabbits in Mexico: a parvovirus. PMID- 2558440 TI - Purification of infectious bronchitis coronavirus by Sephacryl S-1000 gel chromatography. AB - A procedure was developed to purify infectious bronchitis virus (IBV) by gel chromatography (GC) with a Sephacryl S-1000 column. Virus samples concentrated by centrifugation were applied to a Sephacryl S-1000 column and eluted by 0.02 M phosphate buffer (pH 7.2) containing 0.15 M NaCl. Virus particles were recovered mainly in the first peak. Purity of the samples was evaluated by both sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electron microscopy. Using electron microscopy, it was found that there were more spike-rich particles in the virus samples purified by GC than in those purified by sucrose density gradient centrifugation (SDGC). In addition, the hemagglutination unit [log10 (infectivity titer/hemagglutination titer)] of GC-purified virus samples was approximately 10 times lower than that of SDGC-purified virus samples. These results indicate that Sephacryl S-1000 gel chromatography is useful for purification of IBV. PMID- 2558441 TI - Efficacy of a live Pasteurella multocida vaccine for the prevention of experimentally induced bovine pneumonic pasteurellosis. AB - Seventeen Holstein-Friesian calves weighing an average of 139.8 +/- 13.5 (mean +/ standard deviation) kg were used in a study to determine the efficacy of a live vaccine containing of Pasteurella multocida A:3 and Pasteurella haemolytica A:1. Eleven calves received the vaccine by intramuscular injection in the right shoulder, whereas six calves received vaccine diluent and served as non vaccinated controls. Fourteen days following vaccination (Day 15) all calves were inoculated deep intranasally with 3.6 X 10(7) TCID50 bovine herpes virus-1. On Day 16, calves were stressed by transports, and on Day 17 calves were challenged intratracheally with P. multocida A:3. On Day 22 calves were euthanized and necropsied, and tissues were collected for pathological and microbiological evaluations. Scores were assigned to each calf based on the severity of observed clinical signs. Macroscopic lung lesions were expressed as percentage of tissue involved relative to the total lung tissue of a calf. Plasma fibrinogen concentration, rectal temperature, serum antibody level, microscopic appearance of lung, and microbiologic results were also recorded for analyses. The control calves had significantly higher clinical-sign scores (P less than 0.05) and more severe gross lesions (P less than 0.05) than the vaccinated calves. Although the vaccinated calves had a slight increase of immunoglobulins M and G classes, the differences were not statistically significant (P greater than 0.05, P greater than 0.05). The results of the study indicate that the live Pasteurella vaccine is effective against experimental P. multocida infection in calves. PMID- 2558443 TI - [Clinical x-ray and morphological changes in breast carcinomas during radiothermotherapy]. AB - Clinico-roentgeno-morphological investigation established a marked regression of tumor in as many as 86.1% out of 230 patients with stage IIB and III breast cancer who had undergone radiothermotherapy. Morphologically confirmed complete regression was observed in 12.6% of patients with lesions smaller than 3 cm in diameter at presentation. Treatment was followed by massive necrosis, dystrophic changes and inhibition of mitotic activity of tumor cells in patients with stage III cancer and those with stage IIB who had presented with tumors larger than 3 cm in diameter. However, in all these cases viable tumor cells were still observed at the periphery of tumor. PMID- 2558442 TI - Experimental infection of bulls with a genital isolate of bovine herpesvirus-4 and reactivation of latent virus with dexamethasone. AB - Five 13- to 18-month old Belgian Blue bulls were used in this experiment. Four bulls (Nos. 2, 3, 4 and 5) were inoculated intratesticularly with 10(5) plaque forming units of bovine herpesvirus-4 (BHV-4) in each testicle (Day 0). The challenge BHV-4 strain was previously isolated from testicle cells of a bull exhibiting orchitis and azoospermia. The fifth bull (No. 1) was used as a control and received the same volume of uninfected cell culture supernatant. For 5 days, beginning on Day 51 post-infection, two bulls (Nos. 4 and 5) and the control bull (No. 1) received 0.1 mg kg-1 of dexamethasone. Unilateral castrations were then performed at regular intervals for viral examination. Treatment with dexamethasone reactivated latent BHV-4, but no clinical signs were observed in treated bulls until the end of the experiment (Day 93). Only Bull 3 showed conjunctivitis and temporary azoospermia. The virus was recovered from various samples showing that: (i) BHV-4 can be present in a latent state in the testicles and mononuclear blood cells; (ii) dexamethasone reactivates the virus; (iii) the virus is excreted by nasal and ocular routes. Each infected bull seroconverted and a booster antibody response appeared after dexamethasone treatment as shown by immunofluorescence. Neutralizing antibodies were detected in each bull by complement-dependent neutralization test with titres higher than those obtained by a classical neutralization test. No booster response of neutralizing antibodies was observed after dexamethasone treatment. The antigenically relevant envelope BHV-4 proteins were identified by Western blotting using sera samples from the animals. DNA restriction endonuclease profiles of viruses reisolated after primary infection and reactivation showed only small differences. PMID- 2558444 TI - [Induction of renal tumors in monkeys as a result of probable prenatal effects of 1,2-dimethylhydrazine]. AB - Prenatal treatment with 1,2-dimethylhydrazine induced embryonal nephroma in two out of six monkeys. In one case, tumor of the right kidney disseminated to the lower lobes of both lungs. Induced embryonal nephroma of monkey proved similar to Wilms' tumor, a frequent pediatric pathology, in terms of histologic pattern, duration of latent period and clinical course. PMID- 2558445 TI - [The activity of cationic proteins, peroxidase and alkaline phosphatase in the blood neutrophils of diabetics]. AB - Data on the function of neutrophils in diabetes mellitus not infrequently accompanied by inflammatory complications are scarce. The aim of this work is to investigate the state of some components of neutrophil granules (cationic proteins, activity of myeloperoxidase, alkaline phosphatase) that play a major role in the mechanisms of destruction of microorganisms in phagocytosis. PMID- 2558446 TI - [Diseases of the peripheral nervous system in pregnant women]. PMID- 2558447 TI - [Differential diagnostic criteria of blast cells in the peripheral blood in acute myeloid leukemias]. AB - The authors present their data on results of an investigation of blast cells including morphocytochemical examination, determination of the presence of receptors to Fc-fragments of Ig, to C3 component of the complement. Criteria were obtained permitting to differentiate myeloid forms of acute leukosis. PMID- 2558448 TI - [Study of the antiviral activity of nonsteroidal anti-inflammatory agents in cell culture]. PMID- 2558449 TI - [Cellular and humoral immunity in children in cytomegalovirus infections]. PMID- 2558450 TI - [Quantitative relationship between fatty acids comprising lipids of viruses and cells]. PMID- 2558451 TI - [Optimization of conditions of culturing rotaviruses in heteroploid cell lines]. PMID- 2558452 TI - [Optimizing enteral nutrition formulas from the viewpoint of the producer]. PMID- 2558453 TI - Changes in the restriction endonuclease patterns of four modified-live infectious bovine rhinotracheitis virus (IBRV) vaccines after one passage in host animal. AB - Four genomically different bovine herpes virus 1 (BHV 1) vaccines were used to inoculate BHV 1-negative heifers. Restriction endonuclease analysis (REA) of virus isolates from animals during acute infection, after reactivation of virus from latency, and after reactivation followed by superinfection, showed that changes occurred in the BHV 1 genome after one passage in the host animal. Furthermore, the changes varied according to the tissue from which virus was isolated. Southern probe analyses identified the left terminus of the unique long fragment and the inverted repeats as regions of the genome where variability was most often observed. It was concluded that, when REA is used in the diagnostic differentiation of BHV 1 isolates, caution must be applied in the interpretation of results. PMID- 2558455 TI - Malignant brachial artery embolism. AB - We present a rare case of malignant embolization of the brachial artery from a primary pulmonary tumor. The arterial embolism was the initial event in the clinical course of the disease. A review of the literature revealed forty three cases of malignant peripheral arterial embolism reported, and only two similar cases in which the embolization was the initial event of the disease. PMID- 2558456 TI - [The clinical picture and pathogenetic aspects of Kaposi's sarcoma]. PMID- 2558454 TI - Association of steroid therapy with vaccine-associated rashes in children with acute lymphocytic leukaemia who received Oka/Merck varicella vaccine. NIAID Varicella Vaccine Collaborative Study Group. AB - Oka strain varicella vaccine generally has been well tolerated by children with acute lymphocytic leukaemia (ALL) in remission and has induced protection against disease caused by wild-type varicella virus. At the end of 1985, four extensive vaccine-associated rashes were reported among children on maintenance chemotherapy. Steroid therapy in the week before vaccination or in the week following vaccination was significantly associated with rash in a retrospective analysis (odds ratio = 3.84, p = 0.0006). These findings were confirmed prospectively (OR = 2.38, p less than 0.05, one-sided) in a second smaller group of children with ALL on maintenance therapy who received varicella vaccination after the end of the data collection for the initial study but before the relationship between rash and steroids was discovered. As a result of these studies, investigators have been asked to withhold steroids for 1 week before vaccination and to delay resumption of steroid therapy for at least 2 weeks after vaccination. These results should serve as a caution that vaccination of these children should be undertaken only with full knowledge of their therapeutic regimen and a thorough clinical understanding of the competing risks of varicella, vaccination and modification of the child's chemotherapy. PMID- 2558457 TI - [The function of the hypophyseo-adrenal system in patients with disseminated lupus erythematosus]. AB - Radioimmunologic examination of the hypophyseo-adrenal system in patients suffering from disseminated lupus erythematosus with insulin glycemia has revealed changes in the system's reactivity, confirmed by a slow reduction of the blood plasma corticotropin and somatotropin, reaching the initial values by the 120th minute after insulin injection. Decreased functional activity of the adrenal cortex was revealed, manifesting by a slow rise of the blood plasma hydrocortisone by the 30th minute after insulin injection. The examined reserve potentials of the hypophyseo-adrenal system indicate not so much an impairment of the corticotropin secretion central mechanisms, but mostly a decrease of the adrenocortical functional activity. PMID- 2558459 TI - Detailed structural analysis of a genome rearrangement in bovine rotavirus. AB - A genome rearrangement involving RNA segment 11 of a bovine rotavirus has been analysed by molecular cloning and sequencing. This revealed that the rearranged genome segment was generated by a head to tail concatemerisation of two almost full length copies of segment 11. The upstream copy of the gene has lost its 3' end and the downstream copy its 5' end. The truncation of the upstream copy of the gene occurs within the termination codon for VP11 converting it from a UAG to a UGA, the rearranged gene is therefore still able to encode a normal VP11. The possible mechanisms by which this rearrangement may have been generated are discussed. PMID- 2558458 TI - Distinct modes of poliovirus polyprotein initiation in vitro. AB - RNAs of poliovirus type 1 and type 3 were translated in extracts from Krebs-2 cells after annealing with oligodeoxyribonucleotides complementary to different sites in the 5'-untranslated region (5'-UTR). Due to a high level of endogenous RNase H activity in the extracts, such RNAs appeared to be efficiently 5' truncated prior to translation. The observed levels of initiation on differently truncated templates suggested that a region in the middle of the poliovirus 5' UTR is essential for the cap-independent initiation of viral polyprotein synthesis. The data reported here, in conjunction with the results from other laboratories, permitted to relate the essential cis-acting control elements to the 5'-UTR secondary structure domains defined previously (E.V. Pilipenko et al., Virology 168, 201-209). However, the removal of these domains activated another mode of polyprotein initiation, which appeared to require another set of translation initiation factors. PMID- 2558460 TI - The effect of virus-immune hosts on Thogoto virus infection of the tick, Rhipicephalus appendiculatus. AB - Thogoto (THO) virus infections of Rhipicephalus appendiculatus ticks were examined using tick hosts immune to the virus. In the first set of experiments, ticks were infected by feeding on viraemic hamsters. Inter-stadial infection of THO virus was not affected when ticks ingested a virus-immune bloodmeal but there was an effect on persistence of the virus. The incidence of intra-stadial infection was reduced by at least 40% when nymphs partially fed on viraemic hamsters and completed their bloodmeal on a virus-immune guinea pig. When the reverse situation was examined--feeding on a virus-immune host and then a viraemic host--no difference was observed in the number of ticks infected. In the second set of experiments, uninfected ticks acquired virus by co-feeding with infected ticks on apparently non-viraemic guinea pigs. Non-viraemic transmission of the viruses was inhibited when the guinea pigs were immune to either the Sicilian (SiAr 126) or prototype (IIA) isolates of THO virus. The laboratory data indicate that virus-immune hosts may have a significant effect on the role played by ticks in the epidemiology of tick-borne viruses. PMID- 2558461 TI - Budding efficiency of Sendai virus nucleocapsids: influence of size and ends of the RNA. AB - The budding efficiency of Sendai virus antigenomes, as well as of defective interfering (DI) nucleocapsids of the deletion and copy-back types, was compared to that of the viral genome during infections of baby hamster kidney (BHK) cells. The antigenomes were shown to bud into virus particles as efficiently as the genomes, arguing for the irrelevance of the nucleocapsid-RNA ends in regulating the efficiency of budding. The DI nucleocapsids, however, were restricted in their budding by factors inversely proportional to their size, arguing for an effect of nucleocapsid size in this process. This restriction in budding, however, appeared to be only expressed under conditions of very efficient DI-RNA replication. PMID- 2558462 TI - Detection of HSV-1 proteins prior to the appearance of infectious virus in mouse trigeminal ganglia during reactivation of latent infection. AB - Following corneal inoculation of mice HSV-1 produces an acute infection and establishes a latent infection in trigeminal ganglia. The latent virus can be reactivated in vitro by explantation of ganglionic tissue. Viral protein expression was studied in trigeminal ganglia during acute infection of mice and explant reactivation of latent infection. HSV-1 proteins were detectable by immunoprecipitation and immunostaining, in mouse ganglia only from 3-5 days post infection. Although during explant reactivation it has been demonstrated that at 24 h post-explant the trigeminal ganglia are all infectious virus negative (Spivack, O'Boyle II and Fraser (1987) J. Virol. 61, 3288-3291), we have found that three HSV-1 proteins, of 175 kDa, 110 kDa and 90 kDa, are present in latently infected trigeminal ganglia as early as 6-21 h post explantation. Initially, only neuronal cells were positive by immunostaining with anti HSV-1 polyclonal serum for HSV-1 antigens, but at later times HSV-1 antigens were seen in non neuronal cells as well. These proteins may play a role in the initial stages of the reactivation process. PMID- 2558463 TI - Removal of contaminating hemoglobin from peroxidase in traumatic skin lesions. AB - A method is described for the removal of contaminating hemoglobin from the peroxidase enzyme in traumatic skin lesions. The procedure is based on hemoglobin precipitation in a combination of ammonium sulfate half-saturation, and chloroform shaking of the cetyltrimethylammonium-bromide extract. The procedure as such somewhat increases the activity of the peroxidase extract if the extract contains no hemoglobin. On the other hand, the peroxidase activity of the extract decreases as the amount of precipitating hemoglobin increases. On average, about 90% of the peroxidase activity persists after hemoglobin precipitation if the hemoglobin concentration in the extract does not exceed 25 mg/100 ml. In experimental incision wounds, the peroxidase activities obtained with this procedure were the same as when enzyme determinations were done without the removal of hemoglobin or slightly higher. In addition, the amount of peroxidase activity in the wounds was estimated, based on the granulocytes of the contaminating blood. PMID- 2558464 TI - [Structural changes in the dental hard substances and jaw bones of animals due to short-duration weightlessness]. AB - Molars, incisors and parts of corpus mandibulae from a total of 40 male, 30 female (WISTAR SPF), who were exposed for a short-time microgravity, and of their 90 d old offspring were examined. It was calculated the carbonateapatite content in the teeth and in bone in percent by infrared spectroscopy. In terms of quantitative parameters the substances of the space flight group versus the controls varied clearly. Quality and size of these changes depend from sex, investigated material and time of space flight. It can be concluded that an accommodation to the flight stress is possible without any functional disorders also in the oral system. PMID- 2558465 TI - Experimental infection of sheep with bovine leukemia virus: infectivity of blood, nasal and saliva secretions. AB - This study was designed to determine the relative infectivity of lymphocytes and secretions from BLV-infected cattle with and without persistent lymphocytosis (BLV+PL+ and BLV+PL-). Ninety-seven sheep of mixed sex and age were assembled into 21 experimental groups. The recipient sheep were inoculated intravenously with serial dilutions of whole blood, saliva or nasal secretions from BLV+PL+ and BLV+PL- donor cows. Between 200 to 20,000 cells from single and mixed BLV+PL+ or single and mixed BLV+PL- donor cattle were used for inoculation. A very small number of BLV-infected lymphocytes (200 cells) was sufficient to induce BLV infection in sheep inoculated with diluted whole blood from BLV+PL+ cattle. The inoculation of whole blood (containing up to 20,000 lymphocyte cells) from BLV+PL cattle did not induce BLV infection in recipient sheep. Saliva and nasal secretions also failed to bring about BLV transmission. PMID- 2558466 TI - S-phase evaluation with bromodeoxyuridine in lymphocytes from cattle infected with bovine leukemia virus (BLV). AB - Bromodeoxyuridine (BrdUrd), an analogue of thymidine, can be detected by means of monoclonal antibodies and utilized as a marker of the S-phase. In this paper a determination of the S-phase in BLV+ cattle with lymphocytosis has been performed by incorporating bromodeoxyuridine in the DNA. This evaluation was compared to the DNA content, demonstrating that i) bromodeoxyuridine incorporation is a reliable marker of S-phase in BLV+ cattle with lymphocytosis and ii) cytofluorimetry is the method of choice, together with immunocytochemistry, to demonstrate bromodeoxyuridine incorporation. PMID- 2558467 TI - Herpesvirus in harbour seals (Phoca vitulina): isolation, partial characterization and distribution. AB - From post-mortem material (liver and lung) and leucocytes of four (3.6%) out of 112 examined harbour seals during the seal epizootic in 1988 six cytopathogenic viral isolates were obtained which were provisionally classified as herpes-like viruses. Results of physico-chemical and electron microscopic investigations suggested their relationship to the herpesvirus family. Serological examinations were carried out with sera from wildlife as well as captive animals using herpesvirus isolates from four different seals. The neutralization tests revealed as only moderate distribution of seropositive reagents up to 53% of the wildlife seal population. Amongst the seals in the orphanage of Norddeich a very small number of seropositive animals was found. The results obtained indicated a minor role of herpesviruses as primary cause of seal mortality in the North Sea during the 1988 season. PMID- 2558468 TI - Herpesvirus in harbour seals (Phoca vitulina): transmission in homologous host. AB - In the Norddeich orphanage seals about four to five months of age were exposed intranasally and by contact to a phocine herpesvirus which originated from a young seal in this station. Virus excretion by the nasal route was observed in two animals on day 2 and 15 post inoculation, respectively. Production of neutralizing antibodies was detected one week after exposure. The antibody titre range was approximately tenfold higher in the inoculated seals than in animals of the contact group. The seals did not become seriously sick. PMID- 2558469 TI - [The molecular bases of the pathogenesis of AIDS]. AB - The comparative study of the amino acid sequence of gp120 in human immunodeficiency virus (HIV) strains HTLV-III and ARV-2 and the amino end areas of the growth hormone receptors of human skin and the insulin receptors has been carried out, thus making it possible to predict the existence of two compact domains connected with an area of a peptide chain. This area is incapable of the formation of a compact globular structure due to a high content of the remnants of proline. The data obtained as the result of electron microscopic study in combination with image processing have confirmed the predicted three-dimensional structure of gp120. This study has also shown that the amino acid sequence of some regions in the domains of gp120 has a significant degree of homology with similarly located regions of the growth hormone and insulin receptors; in its turn, this amino acid sequence is homologous to the framework regions of the VH domain of immunoglobulin. Antibodies to this VH domain specifically react with recombinant HTLV-III antigen. On the basis of the data obtained in our experiments and from the analysis clinico-immunological information, we have come to the conclusion that AIDS is an autoimmune disease induced by HIV due to the structural homology of gp120 with highly important receptors of human cells. PMID- 2558470 TI - [The epidemiology of rotavirus gastroenteritis]. AB - The materials, comprehensively characterizing the water outbreaks of gastroenteritis with its etiology established and its clinico-epidemiological interpretation made, are presented. Along with other data, the water nature of the outbreak was verified by the detection of rotavirus antigen, made in the enzyme immunoassay, not only in patients, but also in water samples from the surface water source and from sewage. To isolate the infective agent from environmental objects, bentonite, a mineral sorbent, has been used for the first time. The authors believe that the water route plays the main (primary) role in the spread of rotavirus gastroenteritis and the adult population is the main reservoir of this infection. The fact the disease is registered mainly among younger age groups of the child population reflects only the maximum manifestation of rotavirus infection among these age groups. PMID- 2558471 TI - [HLA system antigens in persons with differing susceptibility to the causative agents of acute respiratory diseases]. AB - The relationship between the susceptibility of the body to infections caused by influenza A and B viruses, parainfluenza viruses, adenoviruses, Mycoplasma pneumoniae and antigens of the HLA system was studied on a group of 400 adolescents placed under clinico-epidemiological surveillance for two years. The relationship between histocompatibility antigens and acute respiratory diseases was manifested in a decrease or increase in the occurrence of recurrent diseases and infections or in the probability of the development of the diseases in infected persons. HLA B40 was associated with resistance to influenza A, B18 and B21 were associated with resistance to parainfluenza, B15 and B35 were associated with resistance to M. pneumoniae infection; susceptibility to influenza B was registered in persons with HLA B12 and to M. pneumoniae infection, in persons with HLA B16 and B18. With respect to different infective agents, the relative risk of infection varied within 1.7 and 5.0. PMID- 2558472 TI - [Neurologic complications of myocardial infarction]. AB - Investigated were 275 patients, 30 autopsies and 35 animals with experimental cardiac ischemia and myocardial infarction. Polymorphic neurological complications were detected among which most important were: 1. Cardiocerebral syndromes, crises, strokes; 2. Circulation disorders in anterior and posterior spinal, and posterior radicular arterial beds leading to cardio-spinal crises and spinal strokes; 3. Cardio-brachial, cardio-intercostal, radiculo-cardio-lumbo sacral, cervico-thoracic, brachio-plantar, brachioscapular syndromes. PMID- 2558473 TI - Plasminogen activator and plasmin-like activities in experimental rat tumours. AB - Plasminogen activator activity and plasmin-like amidolytic activity were investigated in two experimental rat tumours, using human plasminogen and chromogenic peptide substrate, S-2251. The invasive hepatocarcinoma and non invasive nephroma were induced with the same chemical carcinogen, dimethylnitrosamine, in F-344 rats and they were continuously transplanted under the renal capsule. While there was no difference in plasmin-like activities of the tumours, the plasminogen activator activity was very low in the nephroma, but high in the hepatocarcinoma. Since the activator activity was completely inhibited by amiloride, it was considered to be of urokinase-type. These results were in accordance with the assumed role of urokinase in the invasion. However, of the respective control organ, kidney was rich in both activities but rat liver contained only very low activities. Therefore the comparison of the plasminogen activator activity of a tumour to the control organ probably does not provide information concerning the malignant transformation as it is suggested in the literature. PMID- 2558474 TI - Cyclic AMP influences protein synthesis in larval brains of Drosophila melanogaster. AB - The protein synthesis in dissected whole larval brains of Drosophila melanogaster has been monitored by [35S] methionine incorporation, as revealed by two dimensional gel-electrophoresis and fluorography. In wild type brains, drugs known to increase cAMP level increased the labelling of at least two proteins in the Mr range 30 to 120 kD and pI range 4.8 to 6.2. One of these proteins, Mr = 78 kD and pI = 5.9, was also enhanced in the dunceM11 memory-mutant, which has an elevated cAMP level, whereas it was hardly affected in the rutabaga memory mutant, which has a subnormal cAMP level. It is suggested that cAMP-induced alterations in protein composition and/or turnover of nerve cells may contribute to the development of memory deficit in the dunce strains. PMID- 2558475 TI - Cation and guanine nucleotide effects on ligand binding properties of mu and delta opioid receptors in rat brain membranes. AB - Monovalent cations such as Na+, K+ and Li+ at 100 mM concentration inhibit the binding of mu and delta opioid agonists (e.g. (3H) dihydromorphine and (3H) D Ala2-Leu5-enkephalin), enhance opioid antagonist [(3H) naloxone] binding in rat brain membranes. Divalent cations have an opposite effect: Mg2+ or Mn2+ (2mM) increase both mu and delta agonist binding, and decrease the antagonist binding. The effect of guanosine 5'-triphosphate and its non-hydrolysable analogue 5' guanylyl-imidodiphosphate at micromolar concentrations is similar to that of sodium ion. In kinetic experiments, guanine nucleotides promote the dissociation of radiolabelled agonists from their binding sites by increasing the rate of dissociation. Equilibrium saturation binding studies show that only binding of the opioid agonist ligands are significantly inhibited by 5'-guanylyl imidodiphosphate. PMID- 2558476 TI - Drugs recently released in Belgium. Lisinopril--ganciclovir. PMID- 2558478 TI - Immunocytochemical localization of the sex hormone-binding globulin in a human hepatoma cell line. AB - The human hepatoma cell line HEP G2 was investigated in an indirect immunofluorescence study for localization of the sex hormone-binding globulin. Cells were grown directly to confluency on the slides used for the immunocytochemical staining. A fine granular cytoplasmatic fluorescence pattern revealed the presence of sex hormone-binding globulin. Chang-liver cells used as controls, incubated under the same conditions, did not fluoresce. The synthesis of the hormone-binding protein in HEP G2 cells, which could be stimulated by L thyroxine in the culture medium, was monitored by either indirect immunofluorescence or by immunoradiometric determination of sex hormone-binding globulin in the supernatant. These studies demonstrate that immunofluorescence techniques are able to detect sex hormone-binding globulin in human hepatoma cells. PMID- 2558477 TI - Insulin-like growth factors I and II in healthy man. Estimations of half-lives and production rates. AB - IGF-I and -II share specific serum carrier proteins which elute on neutral Sephadex G-200 gel permeation chromatography at apparent molecular masses of 50 and 200 kD. The half-lives of free and carrier protein-bound 125I-IGF-I and -II were determined after bolus injections of the tracers into two normal adults. Labelled IGF-I and -II migrated first with the 50-kD and later with the 200-kD complex. In these complexes their apparent half-lives were 20-30 min and 12-15 h, respectively. The apparent half-life a free 125I-IGF-I and -II was 10-12 min. In a second set of experiments, recombinant human insulin-like growth factor I was infused during 6 days in two healthy adults at a dose of 20 micrograms.kg-1.h-1 (corresponding to around 30 mg/day). Serum obtained before and during the infusion was subjected to neutral Sephadex G-200 gel permeation chromatography and fractions were pooled according to the apparent molecular masses at which the carrier protein complexes elute. IGF-I and -II in these pools were determined by RIA. Before the IGF-I infusion, 92 and 272 micrograms/l of IGF-I and -II were found in the 200-kD complex, 45 and 91 micrograms/l in the 50-kD complex, and 15 and 5 micrograms/l were present in the free form. Corresponding figures during the IGF-I infusion were 389 and 18 micrograms/l for the 200-Kd complex, 201 and 54 micrograms/l for the 50-kD complex, and 80 and less than 1 microgram/l for free IGF-I and -II. Using the half-lives of the tracer studies and the levels of the different molecular weight forms of IGF in serum, the production rates for IGF-I and -II were calculated to be 10 mg and 13 mg per day. PMID- 2558480 TI - Codistribution of lectin reactive glycoderivatives and PA-TCH-SP positive sites in rat oviduct. AB - Specimens from rat ampulla and isthmus were stained with a battery of 10 lectin horseradish peroxidase conjugates and lectin binding patterns were correlated with the distribution of periodate-reactive vicinal diol groups as determined by the PA-TCH-SP (periodic acid-thiocarbohydrazide-silver proteinate) sequence. The free surface of ciliated and non-ciliated cells was stained moderately to intensely by all lectins and PA-TCH-SP sequence. Binding sites for WGA, UEA I, LTA, Con A were also detected on the tight junctional regions. Secretory granules reacted with all stainings, except for LPA. The localization of certain sugars (sialic acid, fucose) appeared useful for understanding the functional meaning of positive sites. PMID- 2558479 TI - Relationship of nocturnal plasma bioactive and immunoactive ACTH concentrations to cortisol secretion in normal men. AB - Plasma concentration of ACTH and cortisol were measured at 15-min intervals from 22.30 to 07.00 h in 8 normal male volunteers. ACTH was measured both by a modified and sensitized isolated adrenal cell bioassay and by radioimmunoassay. Within-subject correlations between bioactive ACTH and cortisol concentrations ranged between 0.86 and 0.95, with an average correlation of 0.93 (86% shared variance). In contrast, within-subject correlations between immunoassayable ACTH and cortisol ranged between 0.03 and 0.92, with an average correlation of 0.69 (48% shared variance). These results suggest that plasma ACTH concentrations determined by RIA might not accurately reflect the capacity of plasma to regulate the secretion of adrenal glucocorticoids. PMID- 2558483 TI - Effects of unilateral adrenalectomy on the remaining adrenal cortex of adrenocorticotropic hormone-treated male and female hamsters. AB - Sham-operated and unilaterally adrenalectomized male and female hamsters were administered 25 micrograms adrenocorticotropic hormone (ACTH) for 5 days after the operation in order to examine the effects of ACTH on compensatory adrenal growth. In ACTH-treated male and female hamsters, unilateral adrenalectomy did not change the relative weight of the remaining adrenal. There were no significant differences in the volumes of the adrenocortical zones and their parenchymal cells, as well as in the number of adrenocortical cells per gland if compared with unilaterally adrenalectomized and sham-operated ACTH-treated male hamsters, while 3H-thymidine incorporation per gland was lower in monoadrenalectomized animals. On the contrary, in ACTH-treated females, unilateral adrenalectomy resulted in a significant hypertrophy of zona fasciculata cells and in an enhanced 3H-thymidine uptake by the remaining gland. These findings stress the existence of notable sex-related differences in the compensatory adrenal growth in hamsters. PMID- 2558482 TI - Distribution of intraperitoneal gold colloid (198-Au). AB - The distribution and clearance of intraperitoneally injected 198-Au colloid were investigated in mice. 49% of the radioactivity persisted throughout the first 8 hours after injection and 9% were measured after 7 days. The highest concentrations were then measured within the greater omentum and the mesenteric lymph nodes. PMID- 2558481 TI - Two mechanisms of chromatin compaction. AB - 2 types of chromatin compaction were studied by cytochemical and ultrastructural approach. The 1st type was induced by cultivating transformed cells with dimethylsulfoxide. It caused cell transition into a more differentiated state coupled with the appearance of multiple small chromatin condensates. The 2nd type was induced by nonionic detergent Triton X-100 and/or saline penetration inside the nuclei. It was found to be connected with hypercompaction of chromocenters with their integral areas left unincreased. Both types of chromatin compaction differ in their capacity to induce toluidine blue polymerization on DNA substrate and in stability of DNA towards an early step of acid hydrolysis. The relation of the 2 mechanisms of chromatin compaction to different ways of DNA folding is discussed. PMID- 2558484 TI - Severe neuropathy in a patient with acquired immune deficiency syndrome (AIDS). Evidence for widespread cytomegalovirus infection of peripheral nerve and human immunodeficiency virus-like immunoreactivity of anterior horn cells. AB - A patient with acquired immune deficiency syndrome (AIDS) developed a progressive neuromuscular disorder which included a sensory component, severe weakness and muscle wasting, and fasciculations. At autopsy, there was evidence of severe peripheral neuropathy, as well as widespread cytomegalovirus (CMV) infection within the central and peripheral nervous system. Although the anterior horn cell complement within the spinal cord appeared normal, there was also evidence of human immunodeficiency virus (HIV)-like immunoreactivity of rare anterior horn cells, as judged by immunohistochemical staining. This patient illustrates the complexity of pathogenetic mechanisms operative in AIDS patients with neuromuscular disease, and suggests that at least some examples of neuromuscular disease in patients with this syndrome may be related to widespread CMV infection of the peripheral nerve (including microvascular endothelial cells) and, more rarely, direct HIV infection of some anterior horn cells. PMID- 2558485 TI - Susceptibility of brain cells to murine cytomegalovirus infection in the developing mouse brain. AB - Mouse embryos were infected with murine cytomegalovirus (MCMV) by injecting the virus into the cerebral ventricles in the late stage of gestation; the brains of the offspring were than analyzed using the histological and immunohistochemical methods. Brains of the offspring, which were injected with relatively high titers of MCMV [1 X 10(4) plaque-forming units (pfu)] on day 13 of gestation exo utero or on day 15 of gestation in utero, showed massiv necrosis of the cerebral cortex with gliomesodermal proliferation around 9 to 10 days after birth. In these brains, viral antigen-positive cells were observed in zonal arrangement in the lesion-free cortex and in the hippocampus. Immunohistochemical double staining showed that some of the viral antigen-positive cells had also reacted with antibody to neuron-specific enolase at the same time, but had hardly reacted with antibodies to brain-type creatine kinase or glial fibrillary acidic protein. Brains of the offspring, which were injected with relatively low titers of virus (1 x 10(3) pfu) on day 15 of gestation, showed zonal arrangement of viral antigen positive cells mainly in the cerebral cortex and in the hippocampus 7 days after birth, although the numbers of the positive cells were low. Fourteen days after birth, some of these offspring showed atrophy of the cerebral cortex and the hippocampus. These results suggest that some of the neuronal cells in the cerebral cortex and the hippocampus have special susceptibility to MCMV infection. PMID- 2558486 TI - Peripheral nerve disorders in Lyme-Borreliosis. Nerve biopsy studies from eight cases. AB - Clinical, cerebrospinal fluid and nerve biopsy findings from eight patients with peripheral nervous system complications of Lyme-Borreliosis are reported. Five cases showed the typical features of the Garin-Bujadoux-Bannwarth syndrome (meningoradiculoneuritis), one patient had a multiple mononeuritis associated with acrodermatitis chronica atrophicans Herxheimer. Two cases could not be classified under these diagnostic categories. In all patients we observed a prompt relief of signs and symptoms after antibiotic treatment. Nerve biopsy studies showed gross infiltrations of epineurial vasa nervorum and small infiltrations around endoneurial capillaries. The infiltrations consisted of lymphocytes, histiocytes and plasma cells. We did not find necrotizing changes of the vessel walls, but thrombosis and recanalization was observed in some epineurial vessels. Seven biopsies showed a significant loss of myelinated axons due to axonal degeneration. Only in one biopsy did we observe segmental demyelination next to axonal degeneration. We conclude that the PNS complications of Lyme-Borreliosis in early and late stages of the disease are angiopathic due to vasculitis of the vasa nervorum and primarily caused by axonal degeneration. PMID- 2558488 TI - Neuropathology of gracile axonal dystrophy (GAD) mouse. An animal model of central distal axonopathy in primary sensory neurons. AB - A new neurological mutant mouse shows a gracile axonal dystrophy (GAD). The degenerative lesion develops by postnatal day 80, first appearing in the most rostral portion of the gracile fascicles. This lesion then extends caudally to involve the entire gracile fascicles. Many axonal swellings (dystrophies) also appear in the degenerative lesions in proportion to their severity. The clinical findings develop in keeping with these pathological changes, and are characterized by tremor, ataxia and difficulty in moving the hind limbs. These start around day 80, and progress gradually to death about day 150. The lumbar dorsal roots, their spinal root ganglia and peripheral nerves are normal. Electron microscopic study shows dystrophic axons packed with neurofilaments, mitochondria and tubulovesicular structures. These may reflect some stagnation of axonal transport. The distribution of the lesions suggest that the GAD mouse has a central distal axonopathy involving primary sensory neurons of the lumbar dorsal root ganglia. PMID- 2558487 TI - Cytomegalovirus (CMV) disease of the brain in AIDS and connatal infection: a comparative study by histology, immunocytochemistry and in situ DNA hybridization. AB - Brain tissues from 45 patients with AIDS and two brains with connatal cytomegalic inclusion body disease were investigated for a cytomegalovirus (CMV) etiology of encephalitic lesions. Nineteen brains showed evidence of CMV infection by histology, immunocytochemistry (ICC) using two different antibodies (mono- and polyclonal), and in situ hybridization (ISH). Fourteen cases with typical cytomegalic cells in conventional histology [eight with focally necrotizing encephalitis/ventriculitis including the two connatal infections and six with nodular encephalitis (NE)] revealed CMV with any method. In 5 of 15 AIDS cases of NE without cytomegalic cells, CMV infection was established by ISH, whereas ICC remained negative in these cases. Typical lesions of human immunodeficiency virus (HIV)-induced multi-focal giant cell encephalitis (HIV encephalitis) in 13 brains were never labeled for CMV. In necrotizing encephalitis/ventriculitis, cell types which labeled for CMV, with and without cytomegalic change, comprised neurons, astrocytes, oligodendrocytes, ependyma, choroid plexus, endothelia, and cells in peri- and endoneurium, and in leptomeninges. Both ISH and ICC were able to detect widespread non-cytomegalic CMV-infected cells in normal parenchyma, well beyond the necrotizing lesions, in two AIDS cases. Labeling patterns of nuclei versus cytoplasms varied between the three methods for CMV detection. We conclude that in CNS tissues with cytomegalic cells, ICC and ISH are of comparable sensitivity; however, a diagnosis of CMV disease is possible in such cases by conventional histology. For an in situ diagnosis of CMV infection in NE without cytomegalic cells in AIDS, ISH is the method of choice. A selective vulnerability to CMV infection of any specific cell type of the human CNS is absent.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558489 TI - Cytomembranous inclusions in the peripheral nerves in AIDS. AB - We report finding tubuloreticular inclusions (TRI) in the endothelial cells of endo- and epineurial vessels in the sural nerve of 11 patients with AIDS. Six patients had a painful peripheral neuropathy, one a non-painful sensory neuropathy, one an acute inflammatory demyelinating polyradiculoneuropathy and one a thalidomide-related neuropathy. Two patients had no clinical evidence of neuropathy. The TRI are not specific to one neuropathy and are unlikely to contribute to the pathogenesis of peripheral nerve syndromes in AIDS. PMID- 2558490 TI - Immunoregulation mother-fetus/newborn, a role for anti-idiotypic antibodies. PMID- 2558491 TI - Syndrome of resistance to thyroid hormone in an infant with congenital cytomegalovirus infection. AB - We describe an infant with congenital cytomegalovirus infection who developed the syndrome of resistance to thyroid hormone at three weeks of age, presenting with elevated thyroxine levels and non-suppressible thyroid-hormone-stimulating hormone secretions without any features of thyrotoxicosis. The resistance was present at the peripheral and pituitary levels and resolved spontaneously by 18 months of age. Cytomegalovirus infection was confirmed by the positive urine culture of this virus. This case demonstrates for the first time a possible association of thyroid hormone resistance with congenital cytomegalovirus infection. PMID- 2558492 TI - Chronic Epstein-Barr virus infections associated with coronary aneurysms. AB - We recently reported virological studies in a patient with chronic active Epstein Barr virus (EBV) infection in which T-lymphocytes were infected, and coronary aneurysms were detected by echocardiography. We report here the clinical features of this patient and suggest that EBV may cause coronary aneurysms and that an echocardiographic study should be performed in patients with chronic active EBV infection. PMID- 2558493 TI - Duct-islet cell tumor of the pancreas. A case report with immunohistochemical and electron microscopic findings. AB - A case of pancreatic tumor with features of both duct and islet cell components was found incidentally at autopsy in a 76-year-old male who had died of intrahepatic cholangiocarcinoma. The tumor, measuring about 1.0 cm in diameter, was located in the pancreatic tail. The tumor was composed of two distinct cell populations, islet cells and duct cells. Immunocytochemically, nearly all of the former cells were positive for insulin but negative for cytokeratin, carcinoembryonic antigen (CEA) and mucin, while the latter were positive for cytokeratin, CEA and mucin but negative for insulin. Additionally, a majority of the tumor cells that had formed islet-like structures were positive for neuron specific enolase (NSE), whereas NSE-positive cells were found only rarely in duct components. Electron microscopy confirmed the presence of two cell populations. Simultaneous occurrence of duct and islet cell components in a single pancreatic tumor indicates an intimate histogenetic relationship between pancreatic endocrine and duct cells. PMID- 2558494 TI - [Pharmacological study on a new irreversible agonist of opioid receptors, 7 alpha bis(beta-chloroethyl) aminomethyl-6, 14-endoetheno-tetrahydrooripavine]. AB - 7 alpha-bis (beta-chloroethyl) aminomethyl-6, 14-endoetheno-tetrahydro oripavine (alpha-CAM) is a new irreversible opioid receptor agonist. Its effect on isolated tissues (guinea pig ileum, mouse vas deferens, rat vas deferens and rabbit vas deferens) were studied. It was shown to be bound irreversibly to rat brain P2 membrane preparations. The ED50 of its analgesic effect in mice (icv) was found to be 0.12 nmol/mouse, and the effect may last as long as 2-3 days. It is a compound which produces the longest analgesia known up to date. A single dose (icv) of alpha-CAM was sufficient to produce dependence in mice. Thus, the compound may serve as an agent for studying the mechanism of physical dependence. PMID- 2558495 TI - [Solubilization and characterization of benzodiazepine receptors in frontal cortex of rabbit brain]. AB - [3H]Flunitrazepam binding to benzodiazepine receptors solubilized by the detergent 3-[(3-cholamidopropyl)-dimethyl-ammonio]-l-propanesulfonate (CHAPS) was saturable and showed non-linear Scatchard plot with KD1 0.31 nmol/L and KD2 6.7 nmol/L. The affinities of soluble receptors to benzodiazepine were consistent with P2 membrane. One radioactive zone was found by SDS-PAGE after photoaffinity labelling of soluble membrane and the apparent molecular weight of 55,000 was reported. [3H]Flunitrazepam binding to soluble receptors was enhanced by GABA, NaCl or KCl and barbiturates, but inhibited by bicuculline and picrotoxinin. The enhancement of GABA on [3H]flunitrazepam binding was amplified by NaCl or KCl and antagonized by bicuculline and picrotoxinin. These results suggest that the benzodiazepine receptors solubilized by CHAPS have their pharmacological properties and are still associated with GABA receptors and chloride channel. PMID- 2558496 TI - [Effects of intrahippocampal delta-receptors on inhibition of electroconvulsive shock by electro-acupuncture]. AB - The relationship between the delta-receptors of the hippocampus and electro acupuncture (EA) anticonvulsions was analyzed by radioimmunoassay and microinjection. The content of leu-enkephalin-like immunoreactive substance (LEK LIS) in perfusate from the hippocampus increased after repeated electroconvulsive shock (ECS), while the content of LEK-LIS was not changed after single ECS. EA inhibited epileptiform spikes induced by ECS, and the content of LEK-LIS decreased markedly after EA (30 +/- 9 pg/0.1 ml lower than that before EA). Leu enkephalin (LEK), an agonist of the delta-receptors, 10 micrograms injected into the hippocampus produced epileptiform spikes. However, ICI 174,864 (4 micrograms), a specific antagonist of the delta-receptors, suppressed convulsions significantly. The results suggest that the delta-receptors of the hippocampus participate in EA anticonvulsions. PMID- 2558497 TI - P-7521--a new irreversible opioid ligand. AB - In the receptor binding assay, P-7521 was a potent opioid ligand which acted mainly on mu receptor. The relative affinity ratio at mu, delta and kappa sites was 66:8:1. The inhibitory effects of P-7521 were 1868 and 6060 times more potent than morphine on the electrically evoked contractions in guinea pig ileum and mouse vas deferens, respectively and were readily antagonized by naloxone and Mr2266. These results indicate that P-7521 acted on mu receptor in guinea pig ileum and mouse vas deferens. In rabbit vas deferens, the compound had no agonist activity, but could antagonize the inhibitory effect of U-50488 H, a kappa agonist, showing the antagonistic characterization was on kappa receptor. The dissociation of P-7521 binding to opioid receptor were very difficult in mu binding assay and bioassays. PMID- 2558498 TI - [Effects of tetrandrine on vascular permeability and neutrophil function in acute inflammation]. AB - The effects of tetrandrine (Tet) on vascular permeability and neutrophil (Neu) functions in carrageenin induced subcutaneous air pouch inflammation in rats were studied. It was found that the vascular permeability, Neu emigration, beta glucuronidase (beta-G) release and superoxide anion (O2-) generation were increased in the carrageenin induced inflammation. The vascular permeability, Neu emigration, beta-G release and O2- generation were suppressed by Tet ip (20, 40, 100 mg/kg), but the intracellular superoxide dismutase (SOD) activity and the cAMP level in Neu were increased by the same dose of Tet. The results indicate that Tet inhibits prostaglandin synthesis and scavenges free radicals. The mechanism of the inhibitory effects of Tet on Neu functions may be related to the increases in SOD activity and cAMP levels in Neu. PMID- 2558499 TI - [Inhibition of release of prostaglandins and leukotrienes from calcimycin-induced mouse peritoneal macrophages and bovine aorta endothelial cells by anisodamine]. AB - [3H]Arachidonic acid (AA)-prelabeled mouse peritoneal macrophages were stimulated by calcium ionophore A-23187 to release [3H]AA metabolites. The major labeled products which were co-chromatographed with the authentic PG and LT standards by TLC and determined by liquid scintillation were 6-keto-PGF1 alpha, PGE2, LTC4, and LTB4. Anisodamine (Ani) significantly inhibited the A-23187-induced release of PG and LT from mouse macrophages in a dose-dependent manner. In the presence of Ani at 0.5 mmol/L, the A-23187-induced release of 6-keto-PGF1 alpha, PGE2, LTC4 and LTB4 was reduced by 57%, 20%, 53% and 49%, respectively. A-23187-induced release of 6-keto-PGF1 alpha measured by RIA from bovine aorta endothelial cells was also significantly inhibited by Ani in a dose-dependent manner. These results indicate that the calcium-antagonistic effects of Ani may not only play a significant role in its inhibiting release of PG and LT, but also contribute to its salutary effects in the treatment of septic shock. PMID- 2558500 TI - [Effects of dauricine on the metabolism of arachidonic acid in rat pleural neutrophils]. AB - The effects of dauricine (Dau), an isoquinoline alkaloid and anti-arrhythmic agent used in China recently, on the biosynthesis of metabolites of arachidonic acid in rat pleural neutrophils, comparing with dazoxiben, indomethacin and BW 755 c, were studied. The major products of metabolism by 5-lipoxygenase (5-LPO), measured by HPLC, were LTB4 and 5-HETE, whereas the major cyclooxygenase products measured by HPLC and RIA, were HHT and TXB2. The formation of all products by neutrophils was significantly depressed by Dau in a dose-dependent manner. The concentration of Dau required to obtain 50% inhibition (IC50) of formation of HHT, TXB2, LTB4 and 5-HETE was 25.3, 59.3, 31.8 and 59.5 mumol/L, respectively. These results indicate that the two major metabolic pathways of arachidonic acid in rat pleural neutrophils are inhibited by Dau. PMID- 2558501 TI - Early stages of development of rat brain tumors induced by JC virus: a sequential histological and immunohistochemical study. AB - In order to clarify the origin of JC virus-induced brain tumors in rats, the development of tumors was sequentially analyzed histologically and immunohistochemically. Twenty-two of 30 rats (73%), which were intracerebrally inoculated with JC virus within 24 h of birth (group 1), developed, as a group, 45 brain tumors after 12 to 26 weeks. Seventeen of 27 rats (63%), which were inoculated on the 7th day after birth (group 2), developed 37 brain tumors as a group after a time 12 to 40 weeks. The tumors were found exclusively in the cerebrum. The microtumors, which were defined as tumors less than 2 mm in diameter, were located in the subependymal plate around the ventricular system. The microtumors and most part of the macrotumors consisted of cells of undifferentiated neuroectodermal nature, showing nuclear palisades and Homer Wright-pseudorosette-like structures. Some tumor cells of macrotumors had an astrocytic nature and were positive for glial fibrillary acidic protein, S-100, Leu 7, and vimentin. In conclusion, the target cells of JC virus in rats may be undifferentiated subependymal cells of the cerebrum. The tumor cells show partial glial differentiation as they grow. PMID- 2558502 TI - Rescue of infectious virus from nonproducer Rous cells by chick cellular DNA. AB - The rescue of infectious virus from nonproducer BH RSV(-) cells by chick cellular DNA was attempted in order to investigate the functional state of endogenous and exogenous retroviral genes integrated within the cellular DNA. No infectious virus was rescued by transfection with DNAs of chick helper factor (chf)-negative chick embryo cells (CEC), chf-positive CEC or uninfected CEC producing endogenous Rous associated virus (RAV-0). On the other hand, infectious Rous viruses with the phenotype of RAV-0 and RAV-1 were rescued by transfection with DNAs of CEC which had been infected with RAV-0 and RAV-1. From these results, it seems that exogenous retroviral genes integrated in the cellular DNA are expressed rather easily by transfection while those present endogenously are not. PMID- 2558503 TI - Rapid, simple identification of individual osteoblastic cells and their specific products by cell blotting assay. AB - Biochemical and molecular biological studies of osteoblastic cell function and hormonal regulation are frequently confounded by the inherent cellular heterogeneity and phenotypic instability of existing in vitro and in vivo model systems. A new technique (derived from Western blotting or antibody-based detection of protein molecules bound to nitrocellulose paper) is described for identification of individual cells which synthesize osteoblast-specific gene products (bone Gla-protein, type I collagen, and alkaline phosphatase) or produce cAMP in response to parathyroid hormone (PTH) or isoproterenol. Dispersed primary neonatal rat calvariae or osteogenic sarcoma cells were "plated" on Immobilon-P (a hydrophobic transfer membrane with very high protein-binding capacity) for 30 minutes to several hours, followed by agonist treatment, formalin fixation, hematoxylin staining, and immunostaining with a battery of antibodies specific for osteoblastic products. Individual cells and their secretory zones were visualized by light microscopy and counted. Treatment with PTH with or without isoproterenol resulted in increases in the percentages of osteoblastic cells elaborating cAMP, as well as the intensity of immunostaining, but had no effects on MCF-7 cells, a nonosteoblastic breast carcinoma control line. The percentage of cells within each primary osteoblastic cell population isolated or rat osteogenic sarcoma cell clone (G2 or C12) that elaborated bone-specific proteins or that generated cAMP in response to PTH varied with time and the individual cellular preparation, reconfirming the cellular heterogeneity of these systems. This method, in conjunction with techniques such as in vitro hybridization, should prove useful in characterizing discrete osteoblastic bone cell subpopulations and in clarifying mechanisms of hormonal regulation by local and systemic agents. PMID- 2558504 TI - WR-2721 reduces bone loss after hindlimb tenotomy in rats. AB - WR-2721 is a thiophosphate analog of cysteamine that produces hypocalcemia in vivo. Previous studies suggest that WR-2721 produces hypocalcemia by independent inhibitory effects on parathyroid hormone (PTH) secretion, osteoclastic bone resorption, and tubular reabsorption of calcium. We sought to determine if WR 2721 would decrease bone loss in an animal model of disuse osteoporosis produced by unilateral knee tenotomy in the rat. Tenotomy significantly increased osteoclast number in tibias on the side of the procedure compared with tibias on the opposite side which had not undergone the procedure at 3 and 14 days. Femoral weight of tenotomized limbs were also reduced significantly compared with the contralateral limb at 3 and 14 days. WR-2721 treatment (240 mg/kg daily) prevented 26% of the loss of femoral dry weight and 29% of the loss of femoral ashed weight produced 14 days after tenotomy. In addition, WR-2721 treated (240 mg/kg daily) animals had fewer osteoclasts in tenotomized tibias than control animals at 3 days (6.6 +/- 0.7/mm versus 10.3 +/- 0.9/mm, p less than 0.02) and at 14 days (5.8 +/- 0.3/mm versus 8.7 +/- 0.4/mm, p less than 0.02). These data suggest that WR-2721 decreases bone loss in this model by decreasing osteoclastic bone resorption. PMID- 2558505 TI - Human kallistatin, a new tissue kallikrein-binding protein: purification and characterization. AB - A new and specific tissue kallikrein-binding protein was identified in mammalian serum and in secreted transformed-cell culture media (Chao et al., Biochem. J. 239: 325-331, 1986). We have designated this kallikrein-binding protein as "kallistatin". Human kallistatin has been purified from serum, using chromatographic steps including DEAE-Sephadex, hydroxylapatite, Cibacron blue Sepharose, Sephacryl S200, and preparative polyacrylamide gel electrophoresis. The purified kallistatin consists of a single polypeptide chain with an apparent molecular weight of approximately 54 kDa and isoelectric point of approximately 5.0. Kallistatin was eluted as a single peak on reverse-phase HPLC. The purified kallistatin and 125I-labelled human tissue kallikrein form a approximately a 92 kDa SDS- and heat-stable complex. The complex formation is pH dependent and is inhibited by 0.1% (W/V) of deoxycholate or SDS but not by 0.5% (W/V) of Triton X 100, digitonin, Lubrol or CHAPS. A approximately 54 kDa protein was identified in partially purified kallistatin by polyclonal anti-kallistatin antibodies in Western blot analysis and by its binding to 125I-labelled-human tissue kallikrein in ligand blotting. The role of kallistatin in regulating tissue kallikrein activity and metabolism may now be evaluated. PMID- 2558506 TI - Renal kininases in primary aldosteronism. AB - In order to further clarify the role of renal kallikrein-kinin (K-K) system in primary aldosteronism (PA), daily urinary excretions of renal K-K system components including kallikrein (KAL), kinin (KIN), total kininase (K-ase), K-ase I, K-ase II and neutral endopeptidase (NEP) were measured in PA and normotensives (NT). In this study, a new method for the simultaneous determination of human urinary K-ase I, II and NEP was established and employed. The daily excretions of KAL was significantly higher in PA than that in NT, while no difference was found in KIN between PA and NT. On the other hand, total K-ase in PA (897 +/- 258 micrograms/min/day) was significantly higher than that in NT (209 +/- 6). NEP was also significantly higher in PA (262 +/- 22 micrograms/min/day) than that in NT (127 +/- 6), whereas there were no differences in K-ase I and K-ase II between PA and NT. The relative contributions of K-ase I, II and NEP to total K-ase in NT were 14, 27 and 59%, while those in PA were 12, 17 and 36%, respectively. As a result, these three K-ase contributed only 64% to the total K-ase in PA. These findings suggested that 1) NEP may play a major role in the catabolism of renal KIN in human, 2) NEP is accelerated in PA, 3) unknown K-ase, different from K-ase I, II or NEP, may exist in PA, and 4) accelerated renal K-ase activity may play some role on the disorder of renal water-sodium metabolism and high blood pressure in PA. PMID- 2558507 TI - Monoclonal antibodies against the complex between HMW kininogen and calpain I. PMID- 2558508 TI - A sensitive method for differential determination of kininase I, II and neutral endopeptidase (NEP) in human urine. AB - In order to clarify the significance of NEP in human renal kallikrein-kinin system, an assay system was developed for the simultaneous determination of kininase I, II and NEP activities in human. Each kininase activity was determined by measuring the hydrolysis of bradykinin in the presence of specific inhibitors of kininase I (2-mercaptomethyl-3-guanidinoethylthiopropanoic acid), kininase II (captopril) and NEP (phosphoramidon) in 8 normal subjects. The effects of the different assay buffers on kininase activities were also investigated by using a phosphate buffer. Total kininase, kininase I, II and NEP activities were 499 +/- 65 ng/min/ml (mean +/- S.E.), 55 +/- 8, 141 +/- 21 and 299 +/- 42, respectively in our method using a tris buffer, while a phosphate buffer brought about activities of 358 +/- 43, 45 +/- 5, 156 +/- 21 and 135 +/- 25 ng/min/ml. The relative contributions of kininase I, II and NEP to total kininase activity were 11, 29 and 59% in our assay system, while they were 13, 44 and 35% when a phosphate buffer was used. From these results it was suggested that 1) phosphate may inhibit urinary NEP activity, so that a tris buffer should be used as the incubation buffer, 2) NEP is the major component of human urinary kininases, and 3) NEP may play an important role in the renal kallikrein-kinin system. PMID- 2558509 TI - Localization of neutral endopeptidase in the kidney determined by the stop-flow method. AB - Recently, the existence of neutral endopeptidase (NEP) as a new kininase in the kidney has been reported. In this study, the localization of NEP in the nephron was investigated and compared with other components of the renal kallikrein-kinin (K-K) system by using a stop-flow method in dog kidneys. The stop-flow method was performed according to the procedures previously reported by Scicli et al and Malvin et al. Five mongrel dogs (weighing 15-20 kg) were used in this study. Kininase I, II and NEP were measured by the modified procedure of Ura et al. Kallikrein and kinin were found in the distal tubules, and kininase I and II were observed in both the distal and proximal tubules. NEP was localized mainly in the proximal tubules. A small peak was also recognized in the distal tubules. From these results, it was suggested that, not only kininase I and II but also NEP existing in the proximal tubules may destroy kinin filtered from the glomeruli, and these kininases existing in the distal tubules may play an important role in connection with kinin producing enzymes on the regulation of activity in the renal kallikrein-kinin system. PMID- 2558510 TI - Purification of angiotensin-converting enzyme from human intestine. AB - Angiotensin-converting enzyme (ACE) activity in the intestinal whole homogenate was showed as three peaks on a column of Sephacryl S-300 HR gel filteration. Over 90% of total ACE activity was found in a soluble fraction separated with an ultracentrifuge of the intestinal homogenate, and the ACE activities were detected as two peaks on the same column. On the other hand, two peaks of ACE activities were found in a membrane-bound fraction of treated with trypsin on the Sephacryl column and confirmed with the two peaks of the soluble fraction, while the fraction extracted with Triton X-100 of the membrane-bound fraction showed only one peak as major peak. All ACE peaks were inhibited by addition of EDTA or captopril and by absence of chloride ion completely. We purified one ACE from the soluble fraction by lisinopril-linked Sepharose 6B affinity column chromatography and Cellulofine GCL-200 gel filteration. This enzyme was a 1323-fold purification and its final recovery was 25%. The molecular weight of this enzyme (180,000) was larger than that of ACE from human kidney (170,000), estimated by 7.5% SDS-PAGE. The Km value of the enzyme for HHL was 2.1 mM. The enzyme activity was competitively inhibited by captopril. PMID- 2558511 TI - Distribution of angiotensin I converting enzyme in male reproductive systems of various vertebrates and properties of the genital enzymes. AB - The distribution of the angiotensin I converting enzyme (EC 3.4.15.1, ACE) in male reproductive systems of various vertebrates including non-mammalian species and properties of the genital ACEs were studied. In such mammals as rat, dog and pig, it has been found that ACE activity is only distributed in testis and epididymis (especially in the epididymal semen), but not in accessory sex glands such as prostate, seminal vesicle and coagulating gland. In the rat and dog, most of or all epididymal ACE has been found to resemble testicular ACEs rather than pulmonary ACE in molecular weight. Besides the studies on mammals, it has been found that the enzymes having characteristics of ACE are present in the genital systems of such lower vertebrates as bird (domestic fowl) and fish (carp). PMID- 2558512 TI - Kinin receptors in smooth muscle and vascular effects in sodium depleted rats. PMID- 2558514 TI - Glandular kallikrein, renin and angiotensin converting enzyme of diabetic and hypertensive rats. AB - To clarify the relationship between kallikrein-kinin and renin-angiotensin systems, glandular kallikrein, renin and angiotensin converting enzyme in the submandibular gland, the kidney and plasma were investigated in streptozotocin diabetic and spontaneously hypertensive rats. Kallikrein content in the submandibular gland, the kidney and plasma of diabetic rats was found to be decreased compared with nondiabetic controls. Renin activity in diabetic rats was also reduced in the submandibular gland, but the activity showed no significant changes in the kidney and plasma. The activity of angiotensin converting enzyme (ACE) in plasma significantly increased in diabetic rats. On the other hand, kallikrein content in hypertensive rats was depressed in the kidney, while the content was unchanged in the submandibular gland and plasma. Renin activity in hypertensive rats was found to be higher than that of normotensive rats in the submandibular gland, but the activity showed no remarkable changes in the kidney and plasma. ACE activity in plasma markedly decreased in hypertensive rats in contrast to diabetic rats. In hypertensive-diabetic rats, changes in the levels of these enzymes in tested materials were similar to those of diabetic rats. From these results it is reasonable to assume that (1) reduced kallikrein generation and elevated ACE activity may induce impaired kinin formation and contribute to the development of diabetes mellitus apart from the presence of hypertension and (2) low kallikrein content in the kidney could cause hypertension. PMID- 2558513 TI - Effect of the substance(s) released in vitro by the interaction of Bothrops jararaca (BJ) venom or trypsin and BJ plasma on BJ blood pressure and uterus. PMID- 2558515 TI - Aldosterone, kallikrein, kininase I and II in normal and hypertension complicated pregnancy. PMID- 2558517 TI - Kallikrein-kinin system in newborns of the drug addicted. PMID- 2558516 TI - Kinin-kininase system in drug addict woman in pregnancy and puerperium. PMID- 2558518 TI - Physiologic role of the peripheral enkephalinergic system in regulating cardiovascular homeostasis: evidence of interactions with the renin-angiotensin and kallikrein-kinin systems. AB - On isolated heart preparation, it was found that Leu5-Enkephalin (Leu5-ENK) did not influence the cardiac function. On the other hand, Leu5-ENK induced a specific dose-related inhibition, in the cardiac perfusate, of the activities of kininase II (KII) and angiotensin converting enzyme (ACE) (but not of kininase I KI). Instead no detectable alterations of the above enzymatic activities with the used concentrations of Leu5-ENK were observed in vitro. This opioid also increased specifically the effects induced by some of the autacoids, related to both renin-angiotensin and kallikrein-kinin systems, on the KII and ACE activities. A specific correlation between these Leu5-ENK-induced modifications and the functional responses of the heart to the same autacoids was observed. Naloxone (NAL) and more significantly ICI 174864 (ICI) opposed or reversed the inhibitory effect of the used opioid whereas they had neither inhibitory nor synergic effect on both KII and ACE activity by themselves. The possible physiologic role of the enkephalins in regulating cardiovascular function by acting peripherally on some humoral systems through modulatory mechanism was discussed. PMID- 2558519 TI - Utilization of kininase activities as indicators of malignant disease. PMID- 2558520 TI - Angiotensin converting enzyme inhibitors, captopril and enalaprilat, augment bradykinin-induced prostacyclin synthesis in cultured rat vascular smooth muscle cells. PMID- 2558521 TI - Studies on coagulation-fibrinolysis and kallikrein-kinin systems and kininase activity and kininase II quantity in amniotic fluid. PMID- 2558522 TI - Studies on urinary coagulation-fibrinolysis and kallikrein-kinin systems and kininase in normal pregnancy, labor and puerpreium. PMID- 2558523 TI - Activation of calcium ion-dependent proteinases by bradykinin in dental pulp of the rat. AB - The present study was aimed to examine whether BANA-degrading enzyme activities could be enhanced by bradykinin(BK) in dental pulp of the rat in vitro. The results showed that BK(0.1-10 microM) dose-dependently enhanced BANA-degrading enzyme activity at pH 7.4. The effects of BK(1 microM) were found to be most effective at both pH 7 and 8, with enhancement of the enzyme activities at a wide range of pH. The BK effects at both the pH were not inhibited by FOY-305(0.1 microM), an inhibitor of trypsin-like enzymes, differing from that at pH 6 in adrenal medulla of the rat. On the other hand, the effects of BK at both the pH were remarkably inhibited by EGTA (2 mM), followed by reversal with calcium ion (2.42 mM). These results suggested as follows: 1) there might be two kinds of BANA-degrading enzymes activated by BK in the pulp. 2) it was conceivable that BANA-degrading enzymes activated by BK were quite different from serine proteinases and were interfered with them in the pulp. 3) calcium ion might play a role in BK-induced enhancement of BANA-degrading enzyme activities which were regarded as met-enkephalin (ME) processing enzyme activities in the pulp. PMID- 2558524 TI - Enhancement of primary and secondary immune responses by interferon-gamma. AB - We have investigated the effects of interferon-gamma (IFN-gamma) administered with "G" glycoprotein of vesicular stomatitis virus (VSV), on the neutralizing antibody response. Treatment of mice or cattle with recombinant DNA-derived IFN gamma at the time of primary immunization with "G" glycoprotein enhanced the secondary virus-neutralizing antibody response that followed a booster administration of the same antigen without IFN-gamma treatment. Enhancement was statistically significant, and occurred at relatively low doses of IFN-gamma in the absence of any additional adjuvants. Cattle treated with IFN-gamma at the time of primary immunization were also more resistant to VSV challenge than those immunized without IFN-gamma treatment. Such treatment in conjunction with primary immunization may therefore provide a practical means of enhancing protection from viral challenge without inflammatory adjuvants or boosters. PMID- 2558525 TI - Protection induced by synthetic peptides corresponding to three serotypes in foot and mouth disease virus. PMID- 2558526 TI - Recombinant-derived modified-live herpesvirus vaccines. PMID- 2558527 TI - SIV infected rhesus macaques: an AIDS model for immunoprevention and immunotherapy. AB - SIV infection of macaques constitutes a valuable model for development of AIDS vaccines and antiviral therapies. Initial results using inactivated whole SIV immunogens for immunoprevention and post-infectious immunotherapy in the SIV rhesus monkey system are summarized. PMID- 2558528 TI - Rat liver phosphoribosylpyrophosphate synthetase: existence as heterogeneous aggregates and identification of the catalytic subunit. PMID- 2558529 TI - Regulation of soluble 5'-nucleotidase I from rabbit heart. AB - Rabbit heart contains two soluble 5'-nucleotidases, termed N-I and N-II, which can be separated using phosphocellulose chromatography. N-I prefers AMP over IMP as substrate, in contrast to N-II which prefers IMP over AMP. Both enzymes require Mg2+, but the optimum Mg2+ concentrations for the two enzymes are different. High concentrations of NaCl inhibit N-I and activate N-II. Purified N I is activated by ADP but not by ATP. According to Itoh et al. (1986), purified N II is activated by both ADP and ATP. N-I has been purified approximately 1000 fold to a specific activity of approximately 100 mumol/mg protein/min. The properties of N-I suggest that it is the enzyme responsible for the release of adenosine from AMP under conditions of hypoxia or increased work load. PMID- 2558530 TI - Purine 5'-nucleotidase--its reestimated subunit molecular mass and immunocytochemical localization in chicken liver. PMID- 2558531 TI - The application of affinity chromatography for the separation of "high Km" and "low Km" 5'-nucleotidase and other AMP metabolizing enzymes. AB - AMP-sepharose 4B has been widely used as a general ligand affinity chromatography for purification of AMP deaminase, 5'-nucleotidase, adenosine kinase and other adenine nucleotide metabolizing enzymes. Since these enzymes generally differ in their kinetic properties related to the values of Km for AMP and analogous compounds, it was assumed that there may be a specific elution pattern of some of the enzymes which would enable sequential elution from the column during a single run. Using 0.5 M NaCl, 10 mM ATP and 5 mM adenosine as eluting agents, it was possible to separate on AMP-sepharose column AMP deaminase "high Km" and "low Km" 5'-nucleotidase and adenosine kinase. Adenylate kinase, adenosine deaminase and nonspecific phosphatase did not bind to the column. Using human placental extract, AMP deaminase, "high Km" and "low Km" 5'-nucleotidase and adenosine kinase were purified 2.8, 2.9, 105 and 1240 fold, respectively. AMP deaminase and "high Km" 5'-nucleotidase were further separated using phosphocellulose column chromatography and the final purification was 227 and 143 fold, respectively. The specific activities of purified enzyme preparations were 9.1, 1.0, 0.4 and 0.5 mumols/min/mg protein of AMP deaminase, "high Km" 5'-nucleotidase and adenosine kinase, respectively. This approach provides a rapid method for initial purification of these enzymes from crude soluble extracts. PMID- 2558532 TI - Evidence for "low Km" and "high Km" soluble 5'-nucleotidases in human tissues and rat liver. AB - Two kinetically distinct purine 5'-phosphomono-esterase activities were isolated from soluble fractions of human placenta, cultured human T- and B-lymphoblasts and rat liver using AMP-sepharose chromatography. We have defined these activities as "high Km" and "low Km" 5'-nucleotidase. The relative content of "high Km" and "low Km" activities in the tissues studied ranged from 2 to 264. The optimum pH of "low Km" 5'-nucleotidases ranged from 7.4 to 9.0, Km for AMP from 7 to 15 uM and for IMP from 10 to 26 uM. ATP and ADP were inhibitors of "low Km" enzymes with the apparent Ki values of 55 to 20 uM and 8 to 20 uM for ATP and ADP, respectively. "High Km" 5'-nucleotidases had an optimum pH at 6.5, Km for IMP of 0.3 to 0.5 mM and Km for AMP of 1.0 to 9.4 mM. "High Km" enzymes were activated by ATP with A0.5 values, of 1.7 to 2.3 mM at 100 microM IMP. The data indicate that soluble "low Km" and "high Km" 5'-nucleotidases coexist in mammalian cells and fulfill different functions. These observations suggest a complex system for the regulation of AMP and IMP dephosphorylation. PMID- 2558533 TI - Low 5'nucleotidase activity in mononuclear cells of patients with defect T-cell function. PMID- 2558534 TI - Stimulation by glycerate 2,3-bisphosphate: a common property of cytosolic purine 5'-nucleotidase in various tissues. PMID- 2558536 TI - Anti-5'-nucleotidase antibodies cause human peripheral blood T cells to proliferate. PMID- 2558535 TI - Functional characterization of ecto-5'-nucleotidase (ecto-5'-NT) positive and negative human lymphocytes. PMID- 2558537 TI - Erythrocyte pyrimidine 5'-nucleotidase: HPLC assay, observation on the enzyme behavior. PMID- 2558538 TI - Mechanism of ATP catabolism induced by deoxyadenosine and other nucleosides in adenosine deaminase-inhibited human erythrocytes. PMID- 2558539 TI - Reduction of ara-C cytotoxicity in HL 60 cells by addition of deoxycytidine, cytidine or increased level of cytidine deaminase. PMID- 2558540 TI - The relationship between dihydroorotic acid dehydrogenase and in vitro and in vivo cytostatic effects of brequinar sodium (DUP-785; NSC 368390). PMID- 2558541 TI - Effects of 3'-azido-3'-deoxythymidine on the deoxynucleoside triphosphate pools of cultured human cells. PMID- 2558542 TI - The antiretroviral and cytostatic activity, and metabolism of 3'-azido-2',3' dideoxythymidine, 3'-fluoro-2',3'-dideoxythymidine and 2',3'-dideoxycytidine are highly cell type-dependent. PMID- 2558544 TI - Coordinate regulation of adenosine A1 and A2 receptors. PMID- 2558543 TI - Dideoxycytidine metabolism in wild type and mutant CEM cells deficient in nucleoside transport or deoxycytidine kinase. AB - The growth inhibitory effects and metabolism of 2',3'-dideoxycytidine (ddC) were examined in wild type human CEM T lymphoblasts and in mutant populations of CEM cells that were genetically deficient in either nucleoside transport or deoxycytidine kinase activity. Whereas ddC at a concentration of 4 uM inhibited growth of the wild type CEM parental strain by 50%, two nucleoside transport deficient clones were four-fold resistant to the pyrimidine analog. The deoxycytidine kinase-deficient cell line was virtually completely resistant to growth inhibition by the dideoxynucleoside (ddN) at a concentration or 1024 uM. An 80% diminished rate of [3H]ddC influx into the two nucleoside transport deficient lines could account for their resistance to the ddN, while the resistance of the deoxycytidine kinase deficient cells to ddC toxicity could be explained by a virtually complete failure to incorporate [3H]ddC in situ. Two potent inhibitors of mammalian nucleoside transport, 4-nitrobenzylthioinosine and dipyridamole, mimicked the effects of a genetic deficiency in nucleoside transport with respect to ddC toxicity and incorporation. These data indicate that the intracellular metabolism of ddC in CEM cells is initiated by the nucleoside transport system and the cellular deoxycytidine kinase activity. PMID- 2558545 TI - Purine de novo synthesis and inosinic branch point in vivo in different tissues: a biomathematical model. PMID- 2558546 TI - Use of purine nucleotide and nucleoside metabolising enzymes as tools to determine the presence of purinergic nerve transmission in smooth muscle. PMID- 2558547 TI - The inosinic branch point and its hormonal regulation: evaluation through a mathematical model. PMID- 2558548 TI - Nucleotide pyrophosphatase antagonizes contractions due to the twitch component of the sympathetic nerve response and to applied ATP in the guinea-pig vas deferens. PMID- 2558549 TI - Potent and specific inhibitors of mammalian phosphoribosylpyrophosphate (PRPP) synthetase. PMID- 2558550 TI - Properties of highly purified human T lymphoblast (MOLT-4) deoxycytidine kinase. PMID- 2558552 TI - [A case of giant renal hamartoma considered as congenital mesoblastic nephroma of adult]. AB - A case of giant renal hamartoma considered as congenital mesoblastic nephroma in an adult is reported. A 24-year-old woman was admitted to our hospital with complaints of macroscopic hematuria and left flank pain. On clinical examination, a large left renal tumor was recognized and radical left nephrectomy was performed. The tumor weighed 2,500 g. On histological examination, the tumor was composed of spindle-shaped cells, that were considered as fibroblasts, surrounded by abundant collagenous interstitial material, and many cystic or tubular structures lined by cuboidal epithelium were also seen in the peripheral region. Because of histological similarity we considered this tumor as congenital mesoblastic nephroma in an adult. PMID- 2558551 TI - [Rapid diagnosis of adenoviral conjunctivitis]. AB - Adenoviral conjunctivitis is contagious and it should be diagnosed rapidly. Adenoviral ocular infection showed epidemic keratoconjunctivitis due to Ad8 and pharyngoconjunctival fever due to Ad3 but those clinical pictures are changing because of the appearance of new serotypes of adenovirus such as Ad4, 19 and 37 which are related with acute conjunctivitis. The diagnosis of viral infection is generally time consuming and complex. Enzyme-linked immunosorbent assay (ELISA) and immunofluorescent antibody (IFA) test, using monoclonal antibody to adenovirus hexon have been applied for rapid diagnosis. ELISA showed 75% sensitivity and 100% specificity in 59 cases with adenoviral conjunctivitis and was therefore shown to be more practical than IFA which showed only 67% sensitivity. PMID- 2558553 TI - Diverticular disease of the colon in Ibadan, Nigeria. AB - Eleven cases of diverticular disease of the colon were seen in a review of 603 adult barium enema examinations carried out over a 2-year period (January 1984 December 1985) at the University College Hospital, Ibadan, Nigeria--a prevalence of 1.85%. All the cases were clinically unsuspected and the diagnosis was established only at barium examination. Five of the 11 patients presented with rectal bleeding, six with alteration in bowel habit, six with abdominal pain and associated fever and one with right iliac fossa pain and tenderness mimicking appendicitis. Although an uncommon disease in Nigerians, clinicians are urged to suspect diverticular disease in their differential diagnoses of disorder of the colon in Africans in order not to miss a potentially lethal but treatable condition. PMID- 2558554 TI - The influence of undergraduate clinical training on the attitude of medical students to rural medical practice in Nigeria. AB - A study of a cohort of Nigerian medical students before entry and at the completion of their clinical years of training, has shown that their overall attitude towards rural medical practice deteriorated slightly but not significantly in the process. The attitude had improved slightly in students who had grown up largely in rural areas and deteriorated in those brought up in the townships. Similarly, clinical training did not significantly affect their attitude towards the compulsory 1 year National Youth Service Corps which may involve their service in the rural areas after graduation. Factors which were important in the development of these attitudes, as volunteered by the students, included the area of upbringing (urban or rural), the individual's life plan, and professional exposure before and during training. The importance of the knowledge gained from this study in the educational and health service planning in Nigeria are discussed. PMID- 2558556 TI - Ventilatory pulmonary function study in healthy young Nigerian adults. AB - Ventilatory pulmonary function was studied in 108 healthy young Nigerian adults using an electronic spirometer, with a view to determining the normal values of function for healthy Nigerians. The measured values of forced vital capacity, forced expiratory volume in 1 sec, maximum voluntary ventilation, and peak flow rate were compared with those obtained from similar studies on Nigerians and the predicted values for Caucasians. Higher values of function were obtained in Nigerian males than in the females. Nigerian males, however, showed lower values when compared with Caucasians of similar anthropometric measurements; there was no difference between the females in this study and their Caucasian counterparts. There were also some similarities and differences between the values obtained in this study and those of similar studies in Nigerians of different socio-ethnic backgrounds. The results emphasize the necessity for a ventilatory normogram for each socio-economic or ethnic grouping of individuals. PMID- 2558555 TI - Clinical, hormonal and biochemical features of menopausal women in Ibadan, Nigeria. AB - As a result of the general lack of information about the menopause in Nigeria this study was conducted to determine the prevalence of climacteric symptoms in a cohort of healthy perimenopausal women and also to evaluate the hormonal and biochemical profile of the subjects. Seventeen menopausal and 19 premenopausal women aged 40 years or more were interviewed and had venepuncture and collection of 24-h urine samples. Plasma from the blood samples was used for the assay of LH, FSH, progesterone, 17 beta-oestradiol, calcium and cholesterol while urinary calcium was determined from the 24-h urine samples. The data revealed a significant difference in the prevalence of the symptoms of vasomotor instability (hot flushes, palpitations and excessive sweating) among the menopausal group when compared with the premenopausal group. Furthermore, plasma gonadotrophin levels (LH and FSH) were significantly elevated while plasma oestradiol and progesterone levels were lower in the menopausal women than in the premenopausal group. With regards to the biochemical indices evaluated, only 24-h urinary calcium levels were found to be significantly different for the two groups, with higher levels in the menopausal women. The significance of these findings and the need for adequate screening and treatment of selected menopausal women are discussed. PMID- 2558557 TI - Congenital chest wall malformations in Nigerians. AB - The pattern of congenital chest wall malformations has been studied prospectively at the University of Ilorin Teaching Hospital between 1983 and 1987. A clinical series of 20 patients was found among 2195 patients selected randomly from 10,031 patients obtaining radiographs, and also from 1070 patients seen in the Cardiothoracic Surgery Clinic. An autopsy series of 15 foetuses and newborns was found among 205 studied for malformations. The 35 cases were found to have supernumerary ribs (10 clinical, eight autopsy), absent or defective ribs (three clinical, four autopsy), sternocostal deformities (three clinical, three autopsy) and four clinical cases with thoracic/thoraco-lumbar scoliosis and neonatal dwarfism with short ribs. Apart from two autopsy specimens (Cantrell's pentalogy with acrania and extensive abdominoperineal omphalocele) whose associated malformations appeared to be incompatible with life, all foetuses and newborns apparently had usual perinatal causes of death. One patient presented with costoclavicular syndrome while another had chest wall excision for a suspected tumour in a bifid rib. A patient with Poland's Syndrome and absent digits, and another infant with Cantrell's Syndrome are awaiting surgical reconstruction. The incidence of the malformations is approximately 1.01%. Other clinicopathological features are presented and the literature is reviewed. PMID- 2558558 TI - Schistosomiasis in Zambia: an historical overview and review of the literature. AB - An historical overview and review of the literature on schistosomiasis in Zambia is presented. The review covers a period of approximately 130 years, from Livingstone's exploratory incursion in 1855 to present-day studies. Five species of mammalian schistosomes occur in Zambia and they are reviewed with respect to their distribution, snail hosts, human prevalence, and their role in producing morbidity and disease in man. The present status of infection and disease in Zambia, as well as considerations of transmission and control in the future, are discussed. The references cited represent 90-95% of the published literature on schistosomiasis in Zambia, excluding case reports. PMID- 2558559 TI - Platelet-aggregating activity of released factor(s) from Trypanosoma brucei brucei. AB - The effect of factors derived from Trypanosoma brucei brucei on rat platelets was studied. T. brucei at a concentration of 4 X 10(9) trypanosomes/ml phosphate saline glucose (PSG) was stored at -20 degrees C for 18 h, thawed, and a supernatant fraction, trypanosome-derived supernatant (TDS) was obtained by spinning the sample at 3000 g for 10 min at 20 degrees C. Normal rat platelets, prepared as platelet-rich plasma (PRP), were then incubated with TDS in the absence or presence of ADP (0.05-0.1 microM). The results showed that approximately 83% platelet aggregation was induced by addition of TDS (50 microliters; 113 micrograms protein) to 100 microliters PRP with a platelet count of 10(6). simultaneous addition of ADP and TDS to PRP produced a synergistic effect. It was also shown that a supernatant fraction, obtained by incubating live T. brucei (4 X 10(9)/microliters PSG) at 0 degrees C 1 h and spinning down the trypanosomes (3000 g for 10 min), also induced platelet aggregation. The nature of the factor(s) derived from, or released by, T. brucei inducing platelet aggregation is being investigated but it has been shown not to be ADP. PMID- 2558560 TI - Intestinal parasitic infestation in rural communities: a focus for primary health care in Nigeria. AB - A study carried out in four villages in two local government areas of Oyo State in the southwestern part of Nigeria, revealed that intestinal helminthiasis is still very prevalent in Nigeria. Ascaris is the most common helminth encountered and multiple infestation is also quite common. Schoolchildren and pre-school children are the greatest sufferers of Ascaris, while students and farmers are the greatest target for hookworm. The usefulness of a community-based distribution programme of health services using trained traditional birth attendants and voluntary health workers for data collection and distribution of antihelminth drugs is discussed. PMID- 2558561 TI - Serum copper levels in users of multiload intra-uterine contraceptive devices. AB - The systemic absorption of copper incorporated into multiload intra-uterine contraceptive devices (IUDs), as indicated by serum copper levels in users of such devices, was assessed in a prospective longitudinal study. One hundred and ten healthy Nigerian women using either multiload copper 250 (MLCU 250) or multiload copper 375 (MLCU 375) IUDs participated in the study. Their serum copper levels were estimated serially during 12 months of continuous use of the devices. The mean (+/- s.e.m.) pre-insertion serum copper levels of our subjects using MLCU 250 (17.0 +/- 3 mumol/l) and MLCU 375 (16.7 +/- 0.5 mumol/l) were found to be lower than those reported in Americans (22.2 mumol/l) and in Germans (20.2 mumol/l), although similar to levels in Indians (17.0 mumol/l). There was no significant difference in the mean serum copper levels estimated before and after 1 month of continuous use of the device. Serial estimations of the serum copper levels in users showed that there was no alteration in these levels after a period of 12 months of continuous IUD use. We therefore conclude that the copper incorporated into multiload IUDs appears not to influence the concentration of serum copper of users. PMID- 2558562 TI - Burkitts lymphoma of the ovary in Nigerian adults--a 27-year review. AB - Ten cases of adult Burkitts lymphoma managed over a 27-year period at the University College Hospital, Ibadan were reviewed. The incidence of adult ovarian Burkitts over the period was 1.3%. Menstrual abnormalities and abdominal masses were the most common symptoms while bilateral multinodular ovarian masses were the most constant findings at laparotomy. Diagnosis was mainly by histopathology. Surgery and chemotherapy were the main modalities of treatment. Mortality was high with cerebral metastases being the most common cause of death. PMID- 2558563 TI - Comparative in-vitro antibacterial activity of two brands of antibiotics against clinical isolates of some bacterial genera. AB - Six brands of ampicillin and four of gentamicin were compared for their in-vitro antibacterial activity against clinical isolates of Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa. The minimum inhibitory concentrations obtained for each brand against each bacterial isolate compared very well with one another, and the kinetics of bactericidal activity showed that the brands of each antibiotic possessed similar activity against the clinical isolates tested. PMID- 2558564 TI - Pretreatment urographic evaluation in invasive carcinoma of the cervix uteri. AB - The urographic findings in 52 patients with histologically proven carcinoma of the cervix were studied. Obstructive changes observed were classified according to the severity of calyceal distension and related to the clinical staging of the disease. Obstructive urographic changes were present in 46% of cases, which is three times as high as reported in Caucasian populations, and the severity correlated significantly with the stage of disease. Urography is advocated as a useful pre and post-treatment index of the progress of cervical treatment. PMID- 2558565 TI - 13th Alexander Brown memorial lecture given on 22 February 1985 by G. Onuaguluchi. "Cardiotonic Drugs in the Management of Chronic Congestive Heart Failure: Digitalis Revisited ". PMID- 2558566 TI - Effect of adjuvants on IgG and IgE response to ovalbumin in rats. AB - Specific and total IgG and IgE serum levels were measured in Sprague-Dawley rats after immunization with ovalbumin (OA). OA was administered in an aqueous solution, suspended in alum or in Complete Freund's Adjuvant (CFA) with or without simultaneous administration of Bordetella pertussis (Bp). Control groups received only saline, alum or CFA. Specific IgG, total IgG and total IgE were determined by ELISA methods. Specific IgE was biologically evaluated. The highest specific IgG and IgE antibody responses were obtained with OA suspended in alum, while Bp failed to potentiate this response. The low specific response seen with OA suspended in CFA was potentiated when Bp was simultaneously dosed. Total IgE levels were increased in those groups receiving Bp. The contribution of specific IgG or IgE response to the total IgG or IgE levels was not distinguished with this experimental procedure. It can be concluded that immunization with OA suspended in alum induces appreciable increases in specific IgG and IgE antibodies without significantly affecting total IgG and IgE levels. PMID- 2558567 TI - Dioxins and the Ah receptor. AB - Despite continuing controversies related to public policy, information on the molecular biology of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has advanced significantly over the past decade. Current understanding of the biological mechanisms of TCDD action is based upon the interactions of TCDD with a genetically expressed cytosolic macromolecule that functions as a receptor in many cells across many species. The Ah receptor recognizes TCDD and structurally similar molecules and serves as the transducing step whereby TCDD alters gene expression through the association of the TCDD:receptor complex with specific TCDD-responsive elements on the genome. Understanding these molecular events and their relevance to the organ-level manifestations of TCDD toxicity may be critical to formulating scientifically based assessments of the risk of TCDD exposure. PMID- 2558568 TI - Endothelin, a peptide inhibitor of Na(+)-K(+)-ATPase in intact renaltubular epithelial cells. AB - Endothelin, a potent vasoconstrictor released by vascular endothelial cells, can induce natriuresis in vivo. These studies examined the regulation of Na+ transport by endothelin in suspensions of rabbit proximal tubule (PT) and inner medullary collecting duct (IMCD) cells. Endothelin reduced oxygen consumption (QO2) by 18 +/- 1% in IMCD cells but did not alter QO2 in PT cells. In IMCD cells, endothelin inhibited QO2 half maximally at approximately 5 x 10(-12) M. Several lines of evidence indicate that endothelin reduces QO2 by inhibiting the Na(+)-K(+)-ATPase. 1) Endothelin gave no further inhibition of QO2 after ouabain and blunted the stimulatory effect of amphotericin B on QO2 (+29 +/- 4% in absence of endothelin, 0 +/- 5% in presence of endothelin; n = 6 preparations, P less than 0.001). 2) Endothelin inhibited ouabain-sensitive 86Rb+ uptake by 46.6 +/- 8.6% at 10 s and by 35.4 +/- 5.3% at 30 s without altering uptake at 60 min. 3) Addition of endothelin to IMCD cells induced a net K+ efflux with an initial rate of 32.2 +/- 4.8 nmol.min-1.mg protein-1, consistent with inhibition of the Na(+)-K(+)-ATPase. In contrast to the response observed in intact cells, in permeabilized IMCD cells endothelin did not inhibit ouabain-sensitive ATPase. Several observations indicated that prostaglandin E2 (PGE2) mediates endothelin inhibition of Na(+)-K(+)-ATPase activity. 1) The response to endothelin was blocked by ibuprofen in assays of QO2, net K+ flux, and 86Rb+ uptake. 2) Endothelin and PGE2 gave equivalent, nonadditive inhibition of ouabain-sensitive 86Rb+ uptake.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558569 TI - p-aminohippurate transport in the airways: role of Na+ and HCO3-. AB - The role of Na+ and HCO3- in the transport of p-aminohippurate (PAH) across the canine tracheal epithelium was investigated using Ussing chamber techniques and radiolabeled PAH. Under control conditions, net PAH absorption or a tendency toward net PAH absorption was observed. Neither amiloride (10(-4) M), furosemide (10(-3) M), ouabain (2 x 10(-4) M), nor Na+ substitution of the Ringer solution with choline had any effect on unidirectional PAH fluxes. When the Ringer solution was replaced with a HCO3(-)-free solution, net PAH absorption was consistently observed. In HCO3(-)-free experiments, unidirectional PAH absorptive fluxes were inhibited by mucosal addition of either of the stilbene derivatives, 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS, 10(-4) M) or 4-acetamido 4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS, 10(-4) M). DIDS was more effective than SITS and was also effective in inhibiting PAH absorption in tissues bathed in Ringer solution. Submucosal DIDS or SITS had no effect on PAH fluxes either in HCO3(-)-free or Ringer experiments. We conclude that PAH transport in canine tracheal epithelium occurs by a HCO3(-)-PAH exchange process located on the luminal membrane. PAH transport is not Na+ dependent but is inhibited by both DIDS and SITS. PMID- 2558570 TI - Impaired cell volume regulation in Na(+)-H+ exchange-deficient mutants. AB - To elucidate the mechanism of regulatory volume increase (RVI) in Chinese hamster ovary cells, Na(+)-H+ exchange-deficient mutants, called AP-1, were derived from WT-5 cells, a wildtype subclone. The absence of functional antiports in AP-1 cells was established through measurements of intracellular pH (pHi) and Na+ uptake. Cells exposed to hypotonic medium initially swelled but regained near normal volume within minutes. When isotonicity was then restored, WT-5 cells shrank immediately and then carried out RVI, which was inhibited by 0.1 mM amiloride. This amiloride-sensitive RVI was absent in the AP-1 mutants, suggesting involvement of Na(+)-H+ exchange. In some cell types, RVI is mediated by Na(+)-K(+)-2Cl- cotransport. Bumetanide-sensitive 86Rb+ (K+) influx was detectable in both WT-5 and AP-1 cells, suggesting the presence of Na(+)-K(+)-2Cl cotransport. Bumetanide-sensitive influx was stimulated by osmotic shrinking in WT-5 cells, and only slightly in AP-1 cells. However, Na(+)-K(+)-2Cl- cotransport did not contribute to volume regulation, since bumetanide (50 microM) failed to inhibit RVI in osmotically shrunken WT-5 cells. The inability of cotransport to induce a volume gain in WT-5 cells was attributable to the simultaneous stimulation of Na(+)-K(+)-2Cl- efflux. The rate of efflux was similar in magnitude to the corresponding influx rate so that net Na(+)-K(+)-2Cl- cotransport was negligible. These results show that RVI in osmotically shrunken Chinese hamster ovary cells is mediated by the Na(+)-H+ antiport and that, although stimulated, Na(+)-K(+)-2Cl- cotransport does not contribute to anisosmotic volume regulation. PMID- 2558572 TI - Endothelin-1 activates phospholipase C and mobilizes Ca2+ from extra- and intracellular pools in osteoblastic cells. AB - The effect of endothelin-1 (ET), a novel vasoactive peptide derived from endothelial cells, on osteoblastic MC3T3-E1 cells was studied. ET specifically binds to a single class of high-affinity receptors in MC3T3-E1 cells and induces phospholipase C activation with the production of two second messengers, inositol trisphosphate and 1,2-diacylglycerol, and a biphasic increase in intracellular free Ca2+ concentration ([Ca2+]i), which consists of an initial transient increase and an ensuing sustained plateau, as measured with a fluorescent indicator, fura-2. The second plateau phase but not the initial transient increase in [Ca2+]i induced by ET is abolished by removal of extracellular Ca2+ but not by either nicardipine, verapamil, or diltiazem. The ET-stimulated production of inositol trisphosphate is not abolished by removal of extracellular Ca2+, indicating that ET-stimulated phospholipase C activation is not a consequence of an increase in Ca2+ influx across the plasma membrane. ET causes stimulation of DNA synthesis and reduction of alkaline phosphatase activity in MC3T3-E1 cells. A protein kinase C activator phorbol 12,13-dibutyrate mimics these effects of ET. The results demonstrate that ET activates the inositol lipid signaling pathway and induces mobilization of Ca2+ from both extra- and intracellular pools and activation of protein kinase C in osteoblastic MC3T3-E1 cells. PMID- 2558571 TI - Catecholamine modulation of calcium currents in clonal pancreatic beta-cells. AB - The mechanisms by which norepinephrine and epinephrine activate alpha 2 adrenergic receptors and inhibit insulin release from the pancreatic beta-cell (19, 21, 23) are not yet clear but may involve modulation at several sites. Because intracellular calcium has been implicated in the secretory process, it has been suggested that catecholamines may inhibit secretion by blocking calcium influx, thus reducing the free cytosolic calcium concentration (23). The present study examines the effects of epinephrine, norepinephrine, and clonidine on calcium current in an SV40-transformed hamster beta-cell line (HIT cells). Under voltage-clamp conditions, calcium currents were reversibly inhibited by norepinephrine, epinephrine, and clonidine in the low nanomolar range. The effects were blocked by 1) the alpha 2-antagonist yohimbine, 2) preincubation of the cells with pertussis toxin (PTX), and 3) guanosine 5'-O-(2-thiodiphosphate) (GDP beta S), the nonhydrolyzable GDP analogue that competitively inhibits the interaction of GTP with G proteins. In contrast, guanosine 5'-O-(3 thiotriphosphate) (GTP gamma S) caused irreversible blockade by catecholamines. These effects could not be overcome by adenosine 3',5'-cyclic monophosphate (cAMP), suggesting that the adenylate cyclase pathway is not involved in the G protein coupling with the channels. These studies show that catecholamines inhibit calcium currents in beta-cells through an alpha 2-adrenoreceptor PTX sensitive G protein pathway and could inhibit insulin secretion by this mechanism. PMID- 2558573 TI - Distinct hemodynamic and renal effects of calcitonin gene-related peptide and calcitonin in men. AB - Cardiovascular and renal actions of human calcitonin gene-related peptide II (or beta) (CGRP) and of human calcitonin (CT) infused intravenously for 1 h each (79 and 263 pmol.kg-1.h-1) have been compared in normal men (n = 10 for CGRP, n = 6 for CT and vehicle alone). CGRP lowered diastolic blood pressure by 26% and increased the heart rate by 35% and raised plasma levels of norepinephrine, epinephrine, and dopamine and renin activity (P less than 0.01). The fractional excretion rates (FE) of sodium and chloride were doubled (P less than 0.05-0.01) in the presence of an unaltered glomerular filtration rate. CT, on the other hand, did not affect the diastolic blood pressure, but the stimulation of diuresis and of the FE of sodium and chloride was more pronounced with CT than with CGRP (P less than 0.01). Moreover, CT lowered serum calcium levels and stimulated urinary adenosine 3',5'-cyclic monophosphate and phosphate excretion (P less than 0.01). In conclusion, the cardiovascular effects of CGRP are contrasted by weaker renal tubular actions of the neuropeptide in relation to CT. PMID- 2558574 TI - A1-adenosine receptor-mediated inhibition of adipocyte adenylate cyclase and lipolysis in Zucker rats. AB - Hormone-stimulated lipolysis is reduced in genetically obese rodents and may contribute to the increased adiposity characteristic of the obese state. Endogenously released adenosine, acting via the A1 receptor coupled to the inhibitory guanosine 5'-triphosphate binding protein, Gi, provides a tonic inhibition of lipolysis in rat adipocytes. Removal of this inhibition by the addition of adenosine deaminase frequently results in maximal lipolytic activity. Adipocytes isolated from lean Zucker (Fa/?) rats responded normally to adenosine deaminase, where lipolysis in adipocytes from obese Zucker (fa/fa) rats remained approximately 50% inhibited. Adipocyte adenylate cyclase was equally responsive to activation by forskolin, but lipolytic hormones were significantly less effective in stimulating adenosine 3',5'-cyclic monophosphate (cAMP) production in the obese adipocytes. These cells also exhibited an increased sensitivity to inhibition by the adenosine agonist, N6-(L-2-phenylisopropyl)-adenosine, either in combination with forskolin or beta-adrenergic hormone stimulation. Treatment of isolated adipocytes with pertussis toxin, which uncouples receptor-mediated Gi function, had little effect in cells from lean rats but increased isoproterenol stimulated cAMP production of cells from obese rats to levels observed in the lean cells. In addition, the adenosine A1 antagonist, 8-phenyltheophylline, increased cAMP and lipolytic activity in the obese adipocytes while having little significant effect in the lean adipocytes. These results suggest that hormonal control of lipolysis is altered in the obese Zucker rat because of an alteration in A1-adenosine receptor-mediated inhibition of adenylate cyclase. PMID- 2558576 TI - Estrogens and progestogens conserve bone in rats deficient in calcitonin and parathyroid hormone. AB - To examine the abilities of estrogens and progestogens to slow bone resorption and conserve bone in ovariectomized rats deficient in calcitonin (CT) or parathyroid hormone (PTH), nine groups of animals with 45Ca-labeled bones were studied for 12 wk. Rats were thyroidectomized (TX), parathyroidectomized (PTX), or given sham neck operations (Sham) and treated orally with either estrogen, 300 micrograms 17 beta-estradiol.kg body wt-1.wk-1; progestogen, 500 micrograms norethisterone acetate.kg body wt-1.wk-1; or placebo (Plac). The TX rats had parathyroid autografts and thyroxine replacement. In all surgical groups, estradiol (E2) and norethindrone (Nor) slowed urinary 45Ca excretion and conserved bone (P less than 0.001). However E2 lowered urinary hydroxyproline more than Nor. Total body Ca values (mg +/- SD) were Sham + Plac, 3,079 +/- 201; Sham + E2, 3,886 +/- 335; Sham + Nor, 3,567 +/- 459; TX + Plac, 3,123 +/- 159; TX + E2, 3,869 +/- 235; TX + Nor, 3,540 +/- 422; PTX + Sham, 3,067 +/- 249; PTX + E2, 3,775 +/- 414; PTX + Nor, 3,635 +/- 467. Importantly, E2 and Nor conserved bone as effectively in TX and PTX groups as in Sham rats, although the PTX rats had slower bone resorption and lower plasma 1,25-dihydroxyvitamin D values (P less than 0.001) than groups with intact parathyroids. We conclude that the effects of estrogens and progestogens to slow bone resorption and conserve bone are independent of CT and PTH. These findings appear relevant to the pathogenesis and treatment of postmenopausal osteoporosis. PMID- 2558575 TI - Basolateral and apical binding, internalization, and degradation of insulin by cultured kidney epithelial cells. AB - In vivo, filtered insulin is absorbed and degraded in proximal tubules after binding to the apical membrane. Peritubular removal also occurs and involves basolateral receptor binding and degradation. Whether basolateral degradation proceeds within the cell or on the cell surface is unknown. Because of the difficulties in addressing this question in vivo, this study was carried out with a cultured opossum kidney epithelium cell line with proximal-like features and insulin receptors. Cells were grown in partitioned wells on polycarbonate filters and, when confluent, the monolayer effectively separated the culture well into apical and basolateral compartments. Apical and basolateral binding, internalization, and degradation were studied separately by incubating monolayers with 125I-insulin added to either the apical or basal compartment. At 37 degrees C insulin associated with either pole in a time-dependent manner. This interaction was specific, for it was competitively inhibited by cold insulin but not by unrelated peptides. Separation of surface-bound from internalized insulin was achieved by lowering extracellular pH. At 4 degrees C, 92% of the radioactivity added to either side of the monolayer was surface-bound, whereas at 37 degrees C and after 1 h, 57% was surface-bound and 43% internalized. Affinity of apical and basolateral receptors were similar (1-2 nM), but basolateral receptor number was greater, for at high insulin concentrations (5 x 10(-8) M) basolateral membrane binding exceeded apical by fivefold (250 +/- 81 vs. 56 +/- 11 fm/10(6) cells). Degradation followed exposure to either pole of the cell.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558577 TI - Nonesterified fatty acids modulate steroidogenesis in mouse Leydig cells. AB - The effects of nonesterified fatty acids (NEFA) in modulating testosterone synthesis stimulated by luteinizing hormone (LH, 10 ng/sample) were investigated in isolated adult mouse Leydig cells. LH-stimulated testosterone production was inhibited by triglycerides (0-500 mg/dl, 50 mg/dl, 94% of the control and 500 mg/dl, 40%) and by a mixture of NEFA (100 microM, 70% of control, 200 microM, 54%, and greater than 200 microM, less than 50%). Oleic acid was a more potent inhibitor than linoleic, stearic, or palmitic acids. 8-Bromoadenosine 3',5' cyclic monophosphate (8-BrcAMP, 5 mM) stimulated testosterone production comparable to LH but failed to reverse the inhibition of steroidogenesis produced by the NEFA. The inhibition produced by NEFA was dependent on extracellular Ca2+; and a Ca2+ channel antagonist, verapamil (10 microM), enhanced the inhibition of chylomicrons and fatty acids. 22(R)-hydroxycholesterol (10 microM) reversed the inhibition produced by NEFA. The inhibitory effects of NEFA were reversible by removal of the fatty acids. The results indicate that NEFA are potent modulators of testosterone synthesis in Leydig cells stimulated with either LH, cAMP, or intracellular Ca2+. NEFA inhibit steroidogenesis at one of the steps preceding conversion of cholesterol to pregnenolone. PMID- 2558578 TI - Comparison of the effects of leukotrienes B4 and D4 on distal colonic motility in the rabbit in vivo. AB - Leukotrienes (LTs) have been shown to contract smooth muscle of the small and large intestine in vitro, and an increased mucosal synthesis of leukotrienes has been reported to occur in ulcerative colitis. The purpose of this study was to evaluate the effects of LTs B4 and D4 on the myoelectric and mechanical activity of the rabbit distal colon in vivo and to determine how these effects were modified by indomethacin. LTB4 had a weak excitatory action on both electrical and mechanical activity but this was not statistically significant; LTD4 on the other hand caused a significant dose-related increase in spike potential frequency and contractile activity. Indomethacin alone (1 mg/kg) had no significant effect on the electrical or mechanical activity of the colon but blocked the effects of subsequently administered LTD4. It is concluded from these data that at least part of the excitatory action of LTD4 results from its causing the synthesis and release of excitatory prostaglandins. However, whereas leukotrienes have the potential to play an important role in modulating colonic motility, it seems most likely that this effect would occur after pathological stimulus rather than in the normal healthy colon. PMID- 2558579 TI - Fe-saturation and proteolysis of human lactoferrin: effect on brush-border receptor-mediated uptake of Fe and Mn. AB - We have previously characterized a brush-border membrane receptor that facilitates iron uptake from human lactoferrin. The receptor is specific for human and monkey lactoferrin and does not recognize human transferrin or bovine lactoferrin. In this study, iron uptake from lactoferrin fragments was studied, as well as from lactoferrin partially saturated with iron. Brush-border membrane vesicles (BBMV) prepared from infant rhesus monkey small intestine efficiently accumulated iron from lactoferrin half-molecules, although competition experiments showed that intact lactoferrin has a higher affinity toward the receptor. Lactoferrin partially saturated with iron also effectively delivered iron to the receptor, whereas the affinity was lower than for lactoferrin saturated with iron. Lactoferrin also carries a large proportion of human milk manganese, and receptor-mediated uptake of lactoferrin-bound manganese into BBMV was demonstrated, although this complex had lower affinity than that found for iron-lactoferrin. Thus, although the receptor has a preference for intact iron saturated lactoferrin, partially digested lactoferrin and partially iron saturated lactoferrin can also deliver iron to the receptor. Therefore, these molecular species, which are likely to occur in the gastrointestinal tract of the infant, may contribute to the high degree of iron absorption from human milk lactoferrin. PMID- 2558581 TI - Receptors for extracellular matrix components. AB - The extracellular matrix determines the shape and function of multicellular organisms and permits orderly repair after injury. Although the inherent self assembly properties of many extracellular matrix components such as collagen and elastin foster their deposition and supramolecular organization, cells ultimately dictate the location and composition of regional matrices. These matrices in turn communicate with cells and regulate their attachment, movement, growth, and gene expression. This complex interaction between cells and their matrix is mediated via specific cellular receptors for matrix components, including peripheral and integral membrane glycoproteins, proteoglycans, and glycosyltransferases. This commentary reviews recent advances in the structure and function of receptors for extracellular matrix components. PMID- 2558580 TI - Effect of pH on membrane potential and K+ conductance in cultured rat hepatocytes. AB - We have investigated the effects of extracellular (pHo) and intracellular (pHi) pH on membrane potential difference (PD) and cell conductance (gcell) in rat hepatocytes in primary culture. PD and pHi were measured continuously by using intracellular microelectrodes and the pH-sensitive fluorochrome 2',7'-bis(2 carboxyethyl)-5(6)-carboxy fluorescein (BCECF), respectively, during abrupt changes in the pHo or ionic composition of extracellular perfusate. In the presence of 25 mM HCO3-, PD, gcell, and pHi averaged (+/- SE) -32 +/- 1 mV, 16.4 +/- 1.0 nS, and 7.32 +/- 0.01, respectively. The transference number for K+ (tk+), which reflects the fractional contribution of K+ conductance to gcell, averaged 0.36 +/- 0.03. Exposure to 1 mM Ba2+ produced membrane depolarization and decreased tK+ by approximately 90%. Lowering pHo by a variety of maneuvers in the presence and absence of HCO3- consistently decreased pHi, decreased gcell (approximately 30 nS per unit change in pHi), and depolarized PD. Increasing pHo had opposite effects, but the changes in gcell were generally greater with intracellular acidification than alkalinization. The decrease in PD produced by lowering pHo was associated with a decrease in tK+ of 73 +/- 2% and was inhibited by Ba2+. Exposure to butyrate or withdrawal of NH+4, which lowered pHi without changing pHo, also caused depolarization of PD and a decrease in gcell that was inhibited by Ba2+. These observations indicate that the PD of hepatocytes is strongly influenced by pHi, with or without changes in pHo, and they further suggest that the effects of pH on PD are mediated through changes in plasma membrane K+ conductance. PMID- 2558582 TI - Tumor necrosis factor primes neutrophils for hypochlorous acid production. AB - Tumor necrosis factor (TNF) has a weak direct effect on neutrophil oxidative metabolism and primes neutrophils for oxidant release in response to other stimuli. We examined the effect of recombinant human TNF alpha (rTNF alpha) on production of hypochlorous acid (HOCl) by human neutrophils. TNF alone, even at concentrations of 1,000 U/ml, did not stimulate HOCl production. In contrast, rTNF alpha, in a dose-dependent manner, primed neutrophils for HOCl production in response to the weak agent unopsonized zymosan. rTNF alpha concentrations as low as 10 U/ml resulted in a fivefold increase in HOCl in this system. rTNF alpha primed cells also exhibited increased phagocytosis. Priming in this model system occurred regardless of whether cells were preincubated with rTNF alpha before addition of zymosan or coincubated with both rTNF alpha and zymosan. rTNF alpha priming for HOCl production could not be washed away and required a lag period of approximately 10 min. rTNF alpha priming was not dependent on extracellular Ca2+ and Mg2+. Preincubation experiments demonstrated that rTNF alpha priming was not inhibited by the microfilament blocker cytochalasin B. Although the mechanism remains unclear, these findings demonstrate that rTNF alpha has an important priming effect on the neutrophil myeloperoxidase pathway. PMID- 2558584 TI - [Some pharmacologic comments on the action of glycine derivatives in relation to peculiarities of the central glycinergic system]. AB - The antagonism of the effect of reserpine on the thermoregulation in rats, the potentiation of the reserpine catalepsy with the simultaneous increase of the irritability of rats, the hyperthermia in mice, etc., obtained with some new glycine derivatives, are discussed in relation with recent data on the distribution of the strychnine-sensitive, or strychnine-insensitive-glutamatergic (NMDA) activating, glycinergic receptors. PMID- 2558583 TI - Oxygen tolerance in neonatal rats: role of subcellular superoxide generation. AB - We compared the superoxide anion generating capacity of subcellular fractions from the lungs of neonatal and adult rats. Microsomal and mitochondrial fractions from adult rats produced approximately three times more superoxide (nanomoles per minute per milligram protein) than fractions from neonatal rats in the presence of 100% O2. Subcellular superoxide anion generating capacity was also examined in adult and neonatal rats exposed to greater than 95% fractional concentration of O2 in inspired gas. The O2- produced by mitochondrial and microsomal fractions of adult and neonatal rats increased above control levels for the first 24 h and declined below control values after 48 h of exposure in adults, whereas the elevated O2- production was sustained in microsomal fractions of neonates through 60 h. During the course of hyperoxic exposure, the largest difference in the superoxide generating capacity between adult and neonate was observed after 8-24 h of hyperoxia. The microsomal and mitochondrial fractions from adult rats produced three to seven times more O2- compared with neonatal rats. Cu,Zn superoxide dismutase (SOD) increased during the course of hyperoxia only in neonates at 8, 24, and 48 h of exposure. No change was observed in the activity of Mn SOD. The ratio of SOD activity (units per lung) to subcellular superoxide generating capacity (nanomoles per minute per lung) was calculated for the normal adults and neonates. The ratio for adult rats averaged 23 and 17 for mitochondrial and microsomal fractions, respectively, and 51 for neonatal rats for both subcellular fractions under normoxic conditions. These results suggest that O2- tolerance of neonates may be explained by the favorable balance between antioxidant defenses and subcellular superoxide generating capacity. The role of increased activity of Cu,Zn SOD as an accompanying or a causative phenomenon in O2 tolerance of neonates could not be determined from these experiments. PMID- 2558585 TI - [Effect of glutamic acid palmitamide or linoleamide on convulsions induced by pentetrazole]. AB - Female Balb-C mice received glutamic acid palmitamide (PGt) or linoleamide (LGt) at doses included between 10 and 135 mg.kg-1 (i.p.) and, 90 min after these injections, pentetrazol (PTZ) at 90 mg.kg-1, i.p. PGt (10 or 100 mg.kg-1, i.p.) reduced significantly latencies of first convulsions and lethality and increased the intensity of convulsions after PTZ. This effect disappeared at 135 mg.kg-1, i.p. and, at high doses (up to 150 mg.kg-1, i.p.) PGt antagonized the PTZ convulsions. LGt (10 mg.kg-1, i.p.) had not any significant effect on the PTZ convulsions and antagonized them when administered at a dose of 100 mg.kg-1, i.p. These results, as well as them obtained with PGt and LGt on the haloperidol or reserpine catalepsy, could be the reflection of the preferential impact of LGt in the striatum and of PGt in the limbic structures and in the substantia nigra. At high doses these compounds could reach brain regions with low density of glutamatergic receptors (thalamus, reticular formation) and develop an activating effect able to antagonize the propagation of the PTZ convulsions. These preferential impacts could augur an antiepileptic action for LGT and a promnesient effect for PGt and LGt. PMID- 2558586 TI - [Changes in the strength of the beta-adrenoreceptor inhibiting mechanism on the eve of and during labor determined by the partusisten test]. AB - Partusisten was examined for effects on uterine contractility in 28-36- or 38-42 week pregnant women and parturients. The agent was shown to result in inhibited uterine contractility in the pregnant and parturient females, the contractility was decreased before and particularly during labor, which was indicative of lower beta-adrenoreceptor inhibitory mechanism force. PMID- 2558587 TI - [The importance of rapid methods for the diagnosis of consumption thrombocytopathy in labor]. PMID- 2558588 TI - "The derivative assay"--an analysis of two fast components of DNA rejoining kinetics. AB - The DNA rejoining kinetics of human U-118 MG cells were studied after gamma irradiation with 4 Gy. The analysis of the sealing rate of the induced DNA strand breaks was made with a modification of the DNA unwinding technique. The modification meant that rather than just monitoring the number of existing breaks at each time of analysis, the velocity, at which the rejoining process proceeded, was determined. Two apparent first-order components of single-strand break repair could be identified during the 25 min of analysis. The half-times for the two components were 1.9 and 16 min, respectively. PMID- 2558589 TI - Fourier transform infrared assay of membrane lipids immobilized to silica: leaching and stability of immobilized artificial membrane-bonded phases. AB - A nondestructive, sensitive assay to monitor the hydrocarbon content of silica based chromatography particles has been developed. The assay requires a microscope accessory interfaced with a Fourier transform infrared (FTIR) spectrometer. For determining hydrocarbon content, undiluted alkyl-silica-bonded phases were pressed into a thin wafer. Hydrocarbon content was quantitated using the integrated hydrocarbon band intensity between 2995 and 2825 cm-1 [i.e., band area C-H] and the integrated silica oxide band intensity between 1945 and 1780 cm 1 [i.e., band area Si-O]. Plotting the [band area C-H]/[band area Si-O] ratio vs the carbon content determined by elemental analysis gave a correlation coefficient of r = 0.997. The FTIR assay was validated on 5-, 7-, and 12-microns silica particles using three different immobilized artificial membrane (IAM) silica-bonded phases. The utility of the FTIR assay in determining hydrocarbon content was demonstrated by evaluating hydrocarbon leaching from IAM phases exposed to mobile-phase solvents. The ability of organic solvents to leach hydrocarbon from IAM phases containing phosphatidylcholine (PC) as the immobilized ligand was chloroform greater than ethanol approximately methanol greater than ethyl acetate greater than methylene chloride greater than acetonitrile greater than acetone. Acetone and acetonitrile cause very little hydrocarbon leaching from HPLC-IAM.PC columns. When challenged with different mobile phases, IAM.PC columns perfused with mobile phase are more stable than IAM.PC-bonded phases stirred in mobile phases. IAM.PC contains lecithin linked to silica by amide bonds.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558590 TI - Fluorometric methods employing low concentrations of ethidium bromide for DNA topoisomerase and endonuclease assays. AB - DNA topoisomerase activity can be rapidly assayed by measuring the change in ethidium bromide fluorescence intensity after treatment of closed duplex DNA with enzyme. The sensitivity of the fluorometric assay has been enhanced 3-fold by a 10-fold reduction in ethidium bromide concentration to 0.1 microgram/ml. The results of the fluorometric assays are in close agreement with agarose gel electrophoretic analyses of reacted DNA. A sensitive fluorometric method using 0.1 microgram/ml ethidium bromide has also been developed to determine the fraction of nicked and linear DNAs in a mixture containing closed duplex DNA by measuring the fluorescence intensities of ethidium-DNA complexes at pH 7.0 and pH 12.0. These methods make possible very rapid and sensitive measurements of DNA topoisomerase and endonuclease activities. PMID- 2558591 TI - A micromethod for the assay of cellular secretory physiology: application to rabbit parietal cells. AB - A micromethod for investigating secretory physiology in isolated cells was evaluated. The method utilized a specially designed polycarbonate incubation chamber to provide constant oxygenation to cells incubating in a 96-well microtiter plate. Cells were rapidly separated from media by vacuum filtration. Isolated parietal cells were utilized to demonstrate the versatility of the method for assay of intracellular accumulation of [14C]-aminopyrine, secretion of intrinsic factor into the medium, and assay of intracellular cAMP. Histamine stimulated the uptake of [14C]aminopyrine and intrinsic factor secretion in a sustained and linear fashion. At the end of the 2-h period uptake of aminopyrine and secretion of intrinsic factor were increased 17- and 5-fold, respectively. This response to histamine was accompanied by a rapid and sustained 3-fold rise in intracellular cyclic AMP. In contrast, carbamylcholine caused a transient increase in [14C]aminopyrine accumulation and intrinsic factor secretion which was most pronounced during the first 10 min and had almost ceased by 30 min. Carbamylcholine had no effect on intracellular cAMP levels. This new method, which can handle 400 replicates using parietal cells from the fundic mucosa of a single rabbit, is suitable for studying the time course of intracellular events which accompany general secretory processes. PMID- 2558592 TI - Direct determination of macromolecular charge by equilibrium electrophoresis. AB - Charge is a fundamental property of macromolecules in solution. However, estimation of the apparent charge on polyions has confounded science for decades. Presented here is a general method to determine directly the apparent charge on a polyion, regardless of its size or shape. This new method uses equilibrium electrophoresis, a procedure in which opposing solute flows from electrophoresis and from diffusion balance everywhere as the system reaches a steady-state distribution. The method uses only small quantities of materials, is nondestructive, and requires only simple, inexpensive instrumentation. Here we describe a prototype apparatus, demonstrate the phenomenon, and present experimental examples of the procedure. PMID- 2558593 TI - Bovine testicular beta-galactosidase: purification of enzyme fractions that exhibit high affinity for phosphomannosyl receptors. AB - An improved method is described for the preparation of bovine testicular beta galactosidase that allows the isolation of enzyme fractions that bind avidly to phosphomannosyl receptors. The procedure permits removal of a contaminating beta hexosaminidase and yields nearly homogeneous beta-galactosidase. Enzyme eluted from DEAE-Sephacel was arbitrarily divided into pools that exhibited differing ability to bind phosphomannosyl receptors. A high binding fraction was rapidly assimilated by cultured cells and bound to both low and high molecular weight phosphomannosyl receptors. Carbohydrate analysis of the high binding fraction indicates an average content of one complex and one high mannose oligosaccharide chain per molecule and an average mannose 6-phosphate content of two residues per molecule. However, electrofocusing studies indicated that all the fractions were heterogeneous with respect to sialic acid and phosphate content. The purification procedure also provides highly purified beta-galactosidase suitable for removing beta-galactosidase residues from a variety of complex carbohydrates. PMID- 2558594 TI - Spectrophotometric determination of penicilloates in penicillins. AB - A procedure has been developed for the determination of penicilloates based on the reduction of CuII and detection of CuI using neocuproine. The sensitivity range is 5.6-56 microns penicilloate and the standard deviation was found to be between 1 and 2%. All the penicilloates tested reduced CuII under the conditions of the assay and a slight modification allowed a reasonably accurate determination of the level of this biologically inactive ingredient in a mixture with the parent penicillin. The deterioration of solutions of benzylpenicillin was also studied. PMID- 2558595 TI - A method for kappa-casein genotyping of bulls. AB - A method for kappa-casein genotyping in bulls has been developed. By analysis of DNA polymorphisms we are able to discriminate between the kappa-casein variant A and B in the bulls. This method will be an efficient tool in selection for the most desirable kappa-casein variant. PMID- 2558596 TI - Genetic aspects of bovine leukaemia virus infection and disease progression. PMID- 2558597 TI - Enzyme amplified immunoassays. AB - Enzyme amplification has been applied to improve the sensitivity of many immunoassays which use alkaline phosphatase as the label. The activity of the enzyme is amplified by using NAD, derived by hydrolysis of NADP, to activate catalytically a substrate cycle that generates a coloured product. In routine use, an amplification factor of 100-fold is easily achieved and, with longer incubation times, this can be increased to allow the detection of as few as 3,000 enzyme molecules. In order to avoid the limitations of spectrophotometry, an electrochemical version of the amplifier has recently been developed which performs with comparable sensitivity in a prototype device. The many advantages of electrochemical detection may ultimately herald the arrival of a new generation of simple, sensitive and inexpensive diagnostic systems. PMID- 2558598 TI - The development of a novel immunoassay amplification system and its use in viral detection. AB - A novel amplification system has been developed for the detection of free or antibody-conjugated alkaline phosphatase. The amplification system provides a 100 fold enhancement in the detection of the enzyme, compared to direct detection with chromogenic substrates. The key to the amplification system is the dephosphorylation of a potent phosphorylated inhibitor, and the visualization of this inhibitor using a second, indicator, reaction. This system is shown to provide increased sensitivity for immunoassays detecting either herpes simplex virus or respiratory syncytial virus in clinical samples. In addition, this general concept for amplification may be applicable to a variety of other hydrolytic enzymes, and is demonstrated for the enhanced detection of beta galactosidase. PMID- 2558599 TI - [Synergic effect of 5-fluorocytosine and imidazole derivatives against yeast strains resistant to 5-fluorocytosine]. AB - Antifungal agents associations are widely used in therapy of deep mycotic diseases, particularly amphotericin B-5-fluorocytosine association. Synergistic effect has also been described between 5-fluorocytosine and imidazole derivates. The authors have tested here eventual synergy between 5-fluorocytosine and imidazole derivatives (miconazole, ketoconazole, fluconazole, itraconazole) against 57 years isolates resistant to 5-fluorocytosine by a semi-automated methods in liquid medium (Yeast Nitrogen Base and Brain Heart Infusion). The synergistic effect between 5-fluorocytosine and antifungal imidazoles varies widely with the drug tested. It's more frequent with ketoconazole. Itraconazole and fluconazole present very little synergistic effects in vitro. PMID- 2558600 TI - [Apropos of the monitoring of treatment with low molecular weight heparins]. PMID- 2558601 TI - Genetic and serologic analysis of feline cell-associated herpesvirus-induced infection of the urinary tract in conventionally reared cats. AB - The genetic and antigenic nature of feline cell-associated herpesvirus (FeCAHV) was characterized by use of DNA restriction endonuclease analysis, and direct and indirect fluorescent antibody (FA) techniques. Serologic responses of 6 conventionally reared cats with induced FeCAHV urinary tract infection were retrospectively evaluated, using an indirect FA test. The EcoRI, HindIII, and Pst I restriction endonuclease cleavage patterns of FeCAHV DNA were similar to those of bovid herpesvirus 4 (BHV-4; DN599 strain) DNA. Specific fluorescence was observed when FeCAHV-inoculated cell monolayers were reacted with fluorescein conjugated BHV-4 (DN599 strain) antiserum. Conversely, specific fluorescence was also observed when feline anti-FeCAHV serum and fluorescein-conjugated caprine anti-feline IgG was reacted with BHV-4 (DN599 strain)-infected cell monolayers. At postinoculation week 10, serum antibody titer in cats with FeCAHV-induced urinary tract infection ranged from 1:2,560 to 1:10,240, as measured by use of indirect FA testing. It was concluded that FeCAHV is a member of the BHV-4 group. In addition, the FeCAHV indirect FA test provides a sensitive and specific means of evaluating FeCAHV antibody concentration in exposed cats. PMID- 2558602 TI - Induction of Escherichia coli mastitis in cows fed selenium-deficient or selenium supplemented diets. AB - Ten Holstein heifers were fed a selenium-deficient (SeD) diet (0.04 mg of Se/kg on a total ration dry-matter basis) 3 months before calving and throughout their first lactation. A selenium-supplemented (SeS) diet (2 mg of Se/head/d) was fed to a group of 10 heifers. In about the 14th week of lactation, the cows were challenge-exposed to Escherichia coli by administering 15 to 40 colony-forming units (CFU) into 1 mammary gland. Selenium concentration (microgram/ml) in blood around the time of challenge exposure was 0.033 +/- 0.002 (mean +/- SEM) in SeD and 0.132 +/- 0.006 in SeS cows. Infections were established in all challenge exposed quarters. The frequency of quarter atrophy and agalactia, and reduction in whole-udder milk yield in the first 4 days after challenge exposure, were greater (P less than 0.05) in the SeD cows. Log10 peak bacterial concentrations in milk were higher (P less than 0.05) in SeD (7.63 +/- 0.34 CFU/ml) than in SeS cows (5.57 +/- 0.66 CFU/ml). Mean log bacterial concentration was significantly higher (P less than 0.05) from 12 to 20 hours after challenge exposure in SeD than in SeS cows. Duration of infection was significantly greater (P less than 0.05) in SeD (162.0 +/- 12.0) than in SeS cows (114.4 +/- 18.0 hours). Milk somatic cell counts increased significantly more slowly (P less than 0.05) in SeD than in SeS cows from 8 to 16 hours after challenge exposure. Ratios of milk somatic cells to bacteria in milk were significantly lower (P less than 0.05) in SeD than in SeS cows at 12 and 16 hours after challenge exposure. PMID- 2558603 TI - [Delta agent and hepatocellular carcinoma]. PMID- 2558604 TI - [A new case of Goltz's syndrome]. PMID- 2558605 TI - [Hypothalamic hypophysiotropic neuropeptide receptors]. AB - The present review is dealing with the five major hypothalamic hypophysiotropic neuropeptides (H.H.N.P.) purified and synthesized so far. Four of them specifically stimulate the secretion of one or several anterior pituitary (A.P.) hormones, i.e. thyroliberin (TRH) on TSH and prolactin, gonadoliberin (GnRH) on LH and FSH, corticoliberin (CRF) on ACTH and precursor peptides and somatocrinine (GRF) on GH. The fifth one, somatostatin (SRIF), inhibits the secretion of all A.P. hormones, excepted LH and FSH. All H.H.N.P. affect, positively or negatively, in a dose- and time-dependent manner, the release of stored hormones and their neosynthesis. These responses are submitted to multihormonal modulations. They are initiated by the occupancy of high affinity specific binding sites which have been extensively characterized and morphologically localized. Informations concerning molecular characterization and cloning of receptors for any H.H.N.P. are still awaited. By contrast, the transduction mechanisms which are activated by the occupation of receptors have been extensively studied. They vary depending on H.H.N.P.: TRH and GnRH activate the catabolism of polyphosphoinositides and ensuing pathways, CRF and GRF activate and SRIF inhibits adenylate cyclase dependent pathways. In addition, Ca2+, from extracellular and intracellular sources, play a pivotal role in all cases. The intracellular mechanisms responsible for the last steps of H.H.N.P. action, i.e. exocytosis of secretory granules and transcription of target genes, are however still unknown. PMID- 2558606 TI - [ACTH receptors]. AB - Corticotropin (ACTH) has two main actions in mammalian adrenal cortex: acute stimulation of glucocorticoids secretion and trophic effect which allow the expression of genes encoding for the steroidogenic enzymes. The ACTH membrane bound receptor was one of the first to be demonstrated by direct binding of labeled hormone to subcellular preparations of the adrenal cortex. However, detection and characterization of physiological relevance to ACTH receptors has been difficult, because of the low biological activity of the labeled ACTH. Introduction of a bulky iodine atom into Tyr2 and the oxidation of Met4 appear to contribute most to the loss of activity. These difficulties were overcome recently by using an [125I]-ACTH labeled only in Tyr23, which retains full biological activity. Using this labeled hormone, physiologically relevant ACTH receptors, with high affinity (KD congruent to 10-10M) and low capacity (congruent to 2000 sites/cell) have been characterized in several mammals. A second site of low affinity (KD congruent to 10(-7M) and high capacity, has been found in some studies but the significance of this second site is unknown since it cannot be related to any physiological response of adrenal cells to ACTH. In contrast with the loss of receptors and desensitization of target cells caused by most polypeptide hormones, ACTH seems to regulate positively its own receptors and the cAMP response. The molecular weight of the ACTH receptor appears to be between 83 and 100 KD.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558607 TI - [Glucagon receptors]. AB - Glucagon is a hormonal polypeptide secreted by the A cells of the endocrine pancreas. Its major physiological effects are stimulation of hepatic glycogenolysis and gluconeogenesis. In this review, the current knowledge of receptors and transduction mechanisms involved in the action of glucagon are briefly presented. Receptors and/or an adenyl cyclase system sensitive to glucagon have been identified in the liver, adipocytes, B and D cells of the endocrine pancreas, heart, kidney and brain. In hepatocytes and cytoplasmic membranes of the liver, two populations of receptors with dissociation constants of the oder of 0.1-1 and 10-100 nM respectively have been described. High affinity receptors (10,000-50,000 sites per cell; 2 to 3 pmol/mg of membrane protein) represent approximately 1 to 10% of total receptors. A remarkable property of the glucagon-receptor interaction in the membrane is the decrease in its affinity which can be induced by guanyl nucleotides. Morphologically and biochemically, two events characterise the fate of the glucagon-receptor complex in the hepatocyte: endocytosis of the ligand, and probably the receptor, into an acid cellular compartment and degradation of the ligand. Two of the recently identified degradation products, correspond to sequences 4-29 and 1-13 of the peptide. The major functional consequence of occupation of the receptors is stimulation, via a regulatory protein Gs, of adenyl cyclase activity. More recently, two other effects have been discovered--stimulation of cellular mobilisation of calcium (secondary to an increase in inositol 1,4,5-triphosphate production) and inhibition of the calcium pump leading to an increase in free cytoplasmic calcium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558608 TI - [Cell surface receptors in digestive epithelial cells]. AB - A large number of peptides (neurodigestive peptides) contributes to the regulation of events in the gastrointestinal tract. The demonstration that these mediators interact with receptors as the first step of a direct cellular effect and that the expression receptors at the cell surface is specific for each type of cells and related to the cell function was decisive advances in the understanding of the digestive physiology. Receptors comprise an extracellular ligand binding domain that recognizes specifically each neurodigestive peptide and which is linked to cytoplasmically oriented catalytic domain which transduces the peptide signal and generates a biochemical message. This organization represent a unique system for allosteric modulation of each regulatory biological signal. The modality of the signal activation and control is different for the 4 main families of ligand sensitive receptors linked to 1) the adenylate cyclase activity, 2) the opening of the ionic channels, 3) the activation of the phospholipase C or possessing 4) an intrinsic protein kinase activity. Finally, the transduced message is modulated according to the dynamic state of stimulation or inhibition triggered through the receptor network of the cell and to ratio of cell surface versus internalized receptors. Therefore, the overall functional physiological effect on the digestive cells depends on the integration of the numerous cell surface events induced by the different receptor-activated signaling pathways characteristic of each cell type. In this minireview, the general characteristics of the neurohormonal regulation of the epithelial function and of the ligand receptor interaction, the cartography of the receptors in the different gastric and intestinal epithelial cells their role in the main digestive function (hydroelectrolytic exchanges, cell secretion of products, cellular growth and mitogenis) are briefly exposed and summarized in tables. Recent advances regarding the oncogenic potential of tyrosine kinase receptors in relation to digestive cancer are summarized. PMID- 2558610 TI - [Papilloma of the anus. Management]. PMID- 2558609 TI - [Parathyroid hormone and calcitonin receptors]. AB - Parathyroid hormone and calcitonin are involved in the regulation of calcium metabolism via specific receptors in target organs principally bone and kidney. The pertinent findings in this rapidly evolving area of research are briefly summarized. The receptors for these two hormones have been extensively localised, the transduction of their signal studied, the structure and the proteins composing the receptors characterised by covalent linking and purified by affinity chromatography or specific immunoprecipitation. The sequence of the receptors will be established in a near future using genetic engineering. PMID- 2558611 TI - In vitro activity of amifloxacin against outer membrane mutants of the family Enterobacteriaceae and frequency of spontaneous resistance. AB - Amifloxacin showed potent inhibitory activity against DNA gyrase of Escherichia coli. The difference in the susceptibilities of lipopolysaccharide-deficient Salmonella typhimurium mutants and their parent strain was less than twofold, and the difference in the susceptibilities of porin-deficient E. coli mutants and their parent strain was less than twofold. There was cross resistance among the quinolone group of agents; however, the decrease in MIC for norB mutants was slightly lower than that of other fluoroquinolones. Cell lysis was induced with combined treatment of amifloxacin and sodium dodecyl sulfate in E. coli. The frequency of mutants spontaneously resistant to amifloxacin was extremely low in all species tested. PMID- 2558612 TI - Quinolones and coumarins eliminate chloroplasts from Euglena gracilis. AB - Quinolones and coumarins were potent eliminators of chloroplasts from Euglena gracilis. There was a remarkable similarity between antichloroplastic and antibacterial activities of DNA gyrase inhibitors. Quinolones produced 100% chloroplast-free cells in concentrations which do not affect cell viability. Optimal conditions were exponential growth, continuous illumination, and neutral or slightly alkaline pH. Coumarins were more toxic than quinolones. Among the quinolones, ofloxacin was the most potent in eliminating chloroplasts. Among the coumarins, coumermycin A1 was the most potent. New quinolones and coumermycin A1 were able to induce the complete inability of originally green cells to form green colonies after 24 h of drug exposure, while clorobiocin and novobiocin required several days of exposure. Darkness, heat shock (42 degrees C, 10 min), or simultaneous treatment with chloramphenicol or rifampin decreased the potency of DNA gyrase inhibitors for producing chloroplast-free cells. Remarkably, in cells in which division was blocked by three different methods (resting medium, hyperthermic conditions [37 degrees C], or addition of cycloheximide), new quinolones and coumermycin A1 nevertheless eliminated chloroplasts. The antichloroplastic activity of DNA gyrase inhibitors is additional data suggesting an evolutionary relationship between chloroplasts and eubacteria. PMID- 2558613 TI - Toxicity and efficacy of 2',3'-dideoxycytidine in clinical trials of pigtailed macaques infected with simian retrovirus type 2. AB - Four dosing regimens of 2',3'-dideoxycytidine (ddC) were administered intravenously for 10 to 28 days to 18 pigtailed macaques with simian acquired immunodeficiency syndrome. Ten macaques naturally infected with simian acquired immunodeficiency syndrome retrovirus serotype 2 (SRV-2), the etiologic agent of simian acquired immunodeficiency syndrome, received ddC by continuous intravenous infusion or by a daily bolus injection for 10 to 12 days. Another eight macaques that were negative for SRV-2 and antibody received ddC prophylaxis prior to challenge with virus and continued to receive ddC therapy for up to 28 days postchallenge. All monkeys treated with a continuous intravenous dose of ddC, which maintained plasma concentrations of ddC at levels known to inhibit SRV-2 in vitro, developed dose-related toxic effects, including leukopenia, anemia, lethargy, and decreased appetite. Monkeys treated with a daily bolus injection of ddC experienced more severe toxic effects than those on the continuous intravenous regimen, including exfoliative dermatitis and peripheral neuropathy. At the concentrations of ddC administered, no significant inhibition of SRV-2 replication was detected in naturally infected macaques. However, a prophylactic regimen of ddC did have an inhibitory effect on SRV-2. Our findings suggest that ddC may be valuable as a short-term prophylactic treatment rather than as a long term therapy. PMID- 2558614 TI - Multiplicity of TEM-derived beta-lactamases from Klebsiella pneumoniae strains isolated at the same hospital and relationships between the responsible plasmids. AB - Five plasmid-mediated beta-lactamases conferring high-level resistance to ceftazidime were isolated from Klebsiella pneumoniae strains in the same hospital. These enzymes had isoelectric points ranging from 5.3 to 6.5 (CAZ-1, 5.55; CAZ-2, 6.0; CAZ-3, 5.3; CAZ-6, 6.5; and CAZ-7, 6.3). All isolates and their Escherichia coli transconjugants were highly resistant to amoxicillin (MICs, greater than 4,096 micrograms/ml), piperacillin (64 to 256 micrograms/ml), cephalothin (32 to 256 micrograms/ml), and ceftazidime (32 to 512 micrograms/ml) but remained moderately susceptible to cefotaxime (0.5 to 8 micrograms/ml). Only CAZ-6- and CAZ-7-producing strains were highly resistant to aztreonam (64 to 128 micrograms/ml). All the isolates remained susceptible to moxalactam and imipenem. The reduced activity of piperacillin, cefotaxime, ceftazidime, or aztreonam was restored by 2 micrograms of clavulanate, sulbactam, tazobactam, or brobactam per ml for E. coli producing CAZ-2, CAZ-3, and CAZ-7. Sulbactam had a lower protective effect than other inhibitors for E. coli harboring CAZ-1 and especially CAZ-6. Except for CAZ-1, which was mediated by a 150-kilobase (kb) plasmid (pCFF14), the other ceftazidimases were mediated by plasmids of 85 kb with EcoRI digestion patterns similar to that of pCFF04 encoding CTX-1 beta lactamase. A TEM probe hybridized with a 19-kb EcoRI fragment of all these closely related plasmids. PMID- 2558615 TI - Comparative in vitro and in vivo activities of piperacillin combined with the beta-lactamase inhibitors tazobactam, clavulanic acid, and sulbactam. AB - Tazobactam (YTR-830H), a novel beta-lactamase inhibitor, was compared with clavulanic acid and sulbactam for enhancement of the activity of piperacillin against beta-lactamase-producing, piperacillin-resistant clinical isolates. Piperacillin MICs were determined in media containing a fixed concentration of 2 or 4 micrograms of the inhibitors per ml. The higher concentration was generally more effective. Tazobactam was superior to sulbactam in enhancing the spectrum and potency of piperacillin. Although the calvulanic acid combination was more potent, tazobactam was effective for a similar spectrum of resistant gram negative clinical isolates containing beta-lactamase. MICs were reduced to the susceptible range for Escherichia coli, Klebsiella pneumoniae, Proteus spp., Salmonella spp., and Shigella spp. Combinations with tazobactam and sulbactam, but not clavulanic acid, were effective against Morganella spp. Some antagonism of the activity of piperacillin was observed with clavulanic acid but not with tazobactam or sulbactam. The inhibitors were similarly effective with piperacillin against beta-lactamase-positive Staphylococcus spp. and the Bacteroides fragilis group. Piperacillin-tazobactam was more effective against a broader spectrum of gram-negative enteric bacteria than ticarcillin plus clavulanic acid was. Combinations with tazobactam or clavulanic acid had a broader spectrum of activity than combinations with sulbactam against bacteria that produce characterized plasmid-mediated enzymes of clinical significance. In particular, piperacillin with tazobactam or clavulanic acid, but not with sulbactam, inhibited TEM-1, TEM-2, and SHV-1 enzymes. In vitro activity was reflected in vivo. Tazobactam and clavulanic acid were superior to sulbactam in enhancing the therapeutic efficacy of piperacillin in mice infected with beta lactamase-positive E. coli, K. pneumoniae, Proteus mirabilis, and Staphylococcus aureus. Only combinations with tazobactam and sulbactam were effective against the Morganella infection. Tazobactam has a good potential for enhancing the clinical efficacy of piperacillin. PMID- 2558616 TI - Combined antibody and ganciclovir treatment of murine cytomegalovirus-infected normal and immunosuppressed BALB/c mice. AB - The efficacy of treatment with ganciclovir (DHPG) and antibody activity containing ascitic fluid (AF) separately and in combination was studied in normal and immunosuppressed BALB/c mice challenged intraperitoneally with a lethal dose (10(6) PFU) of murine cytomegalovirus (CMV). With combination therapy, lower doses of both DHPG and AF were often as effective as a higher dose of either agent given singly. For instance, the survival rate of murine CMV-challenged immunosuppressed mice was doubled when 4 mg of DHPG per kg and a 1:16 dilution of AF were both administered in contrast to when each was used alone. In both groups of animals, combination therapy was shown to be more effective than either therapy individually, even when initiation of therapy was delayed as long as 48 h. Such an approach holds promise for decreasing the expense associated with antibody use and the dose-related toxicity associated with DHPG use while maintaining or possibly increasing the efficacy of prophylaxis and therapy of serious CMV disease in humans. PMID- 2558617 TI - In vitro activity of AT-4140 against clinical bacterial isolates. AB - The activity of AT-4140, a new fluoroquinolone, was evaluated against a wide range of clinical bacterial isolates and compared with those of existing analogs. AT-4140 had a broad spectrum and a potent activity against gram-positive and negative bacteria, including Legionella spp. and Bacteroides fragilis. The activity of AT-4140 against gram-positive and -negative cocci, including Acinetobacter calcoaceticus, was higher than those of ciprofloxacin, ofloxacin, and norfloxacin. Its activity against gram-negative rods was generally comparable to that of ciprofloxacin. Some isolates of methicillin-resistant Staphylococcus aureus (MIC of methicillin, greater than or equal to 12.5 micrograms/ml) were resistant to existing quinolones, but many of them were still susceptible to AT 4140 at concentrations below 0.39 micrograms/ml. The MICs of AT-4140, ciprofloxacin, ofloxacin, and norfloxacin for 90% of clinical isolates of methicillin-resistant S. aureus were 0.2, 12.5, 6.25, and 100 micrograms/ml, respectively. AT-4140 was bactericidal for each of 20 clinical isolates of Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Serratia marcescens, and Pseudomonas aeruginosa at concentrations near the MICs. AT-4140 inhibited the supercoiling activity of DNA gyrase from E. coli. PMID- 2558618 TI - DNase activity of micrococcal endonuclease insolubilized on corn cob. AB - The endonuclease from S. aureus has been immobilized on ground maize cob, previously activated with tosyl chloride. Pretreatment of the support on acid before tosylation yielded the best insoluble enzyme derivatives. The catalytic activity has been evaluated as percent of total hydrolysis attained in a batch reactor using DNA as a model substrate. The derivatives prepared are very resistant to high temperatures under conditions of catalysis (24 h at 45 degrees C). For these long reaction times, the extent of hydrolysis in the presence of small amounts of organic solvent (dimethyl sulfoxide at 2 percent) is larger than in plain buffer (Tris). This type of derivative could be very useful for the removal of nucleic acids from single-cell protein concentrates. PMID- 2558619 TI - Transendothelial cell diapedesis of neutrophils in inflamed human skin. AB - The mode of extravasation of neutrophils (PMNs) in cutaneous inflammation was studied in sequential biopsy specimens taken from human skin. Inflammatory skin reactions were produced by intracutaneous injection of endogeneous mediators of inflammation--C5ades arg, LTB4, neutrophil-activating peptide (NAP) and interleukin-1 (IL-1). Within 30 min after injection neutrophils were observed in close contact with endothelial cells of postcapillary venules and, following cytoplasmic engulfment, the cells were found to be transported transcellulary through the endothelial layer. In a total of 20 biopsy specimens taken at various times, cell migration via interendothelial gaps was absent. Instead, the transcellular pathway appeared to be the first and foremost mode of diapedesis. During this migratory process PMNs lacked signs of degranulation and numerous electron-lucent vesicles and secondary lysosomes were found. In addition, coated pits present on leukocyte as well as endothelial-cell membranes were indicative of receptor-mediated endocytotic processes. PMID- 2558620 TI - [Problems of the molecular mechanism of the effects of synthetic antiviral drugs]. AB - Results of experimental study on the effect of bonafton, tebrofen, florenal and riodoxol on synthesis of virus specific proteins and RNA are presented. Selective inhibition of RNA-transcripts and respective proteins under the effect of bonafton was revealed. Florenal had a selective inhibitory effect on synthesis of virus specific proteins. Riodoxol did not inhibit synthesis of viral RNA and proteins of the influenza virus. PMID- 2558621 TI - [Genital human papillomavirus infections in certain groups of women in the Antwerp region]. AB - Different groups of women, living in the Antwerp region, are examined for the occurrence of genital human papillomavirus (HPV)-infections. To detect these infections, physical examination and DNA-DNA-hybridisation technics (Southern Blotting with the probes of HPV 6, 11, 16 and 18) have been used. Part of these women are also examined colposcopically. The following groups of women have been included in the study: --women who present for cervical cancer screening with their general practitioner after a communal campaign. (HPV detection by physical examination, cytology and hybridisation); --women who present for cervical cancer screening with the mobile team after a communal campaign. (HPV detection by physical and colposcopical examination, by cytology and by hybridisation); - women who are routinely screened for cervical cancer in a gynecological private practice, situated in the same region. (HPV detection by physical examination and hybridisation). Prevalences of HPV infections in these groups will be presented and the detection methods used will be discussed. PMID- 2558622 TI - [Evaluation of the effectiveness of a cicletanine-enalapril combination in hypertensive patients]. AB - In this multicentre controlled single blind trial the effectiveness and safety of cicletanine (100 mg/day) were compared with those of enalapril (20 mg/day) and of the combination of both drugs in the same doses in 72 patients (41 men, 31 women, mean age 64.1 +/- 8.3 years) with permanent moderate essential hypertension without severe cardiovascular complications. In the course of the trial, one patient in each of the three therapeutic groups was excluded either for insufficient effectiveness in monotherapy or for photosensitization under the combined treatment. After two months of treatment, the fall in blood pressure and the number of patients with normalized BP were similar in the groups treated with cicletanine or enalapril alone. In contrast, the cicletanine-enalapril combination produced a significantly greater fall of diastolic arterial pressure than cicletanine alone. In addition, there was a greater reduction of functional symptoms associated with arterial hypertension. Apart from the lone case of photosensitization observed with the combined treatment, only minor side-effects were encountered, including an episode of diarrhoea and a case of extrasystoles with the combination, and a case or nausea with lipothymia under cicletanine alone. There were no significant variations of biochemical values. PMID- 2558623 TI - Cicletanine stimulates cholesteryl ester hydrolase activities and prostacyclin production in arterial smooth muscle cells. AB - Cicletanine (1,3,-dihydro-6-methyl-7-hydroxy-3-(4-chlorophenyl) furo (3,4 c)pyridine) is a novel antihypertensive agent whose principal mechanisms of action may be stimulation of arterial PGI2 synthesis. Since PGI2 and related eicosanoids promote cholesteryl ester hydrolytic activities in arterial smooth muscle cells via cyclic AMP, we evaluated the effects of cicletanine on PGI2 production, the enzymes comprising the arterial cholesteryl ester cycle, and cyclic AMP generation by arterial smooth muscle cells in tissue culture. Toxicity was monitored using dye-exclusion tests. Monocyte adhesion essays were also performed to determine if cicletanine would make blood-derived monocytes less adherent to endothelium, thus less prone to initiating a pre-atherosclerotic event. Cells were exposed to a standard dose of 1.7 mM cicletanine for 2 hours. Cicletanine stimulated PGI2 production 2-3 fold over basal levels. Furthermore, cicletanine stimulated both lysosomal and cytoplasmic CE-hydrolases, with modest inhibitory effect on CE synthesis. However, cyclic AMP was not increased following cicletanine treatment. These data suggest that cicletanine may stimulate CE-hydrolase activity directly, and independently of endogenously synthesized eicosanoids. These effects were not due to toxic effects of this compound, as determined by dye exclusion assays. No effects were seen on monocyte adhesion to endothelial cells in vitro. These results suggest that in addition to being a novel, potent antihypertensive agent, cicletanine may also contribute to cholesterol mobilization in smooth muscle cells by stimulating CE hydrolysis. PMID- 2558624 TI - Characterization of phosphoenolpyruvate synthase mutants in Salmonella typhimurium. AB - The enteric bacteria are able to grow by utilizing three-carbon compounds (pyruvate, lactate, and alanine) as sole carbon sources only if they have a functional phosphoenolpyruvate synthase (PEP synthase). PEP synthase catalyzes the phosphorylation of pyruvate to PEP with the hydrolysis of ATP to AMP. This anaplerotic reaction is needed for the synthesis of carbohydrates and citric acid cycle intermediates that are essential for continued cell growth. Insertion mutants were isolated in Salmonella typhimurium that specifically lack the ability to grow on three-carbon compounds. These mutants also fail to utilize acetate as a sole carbon source. Enzyme assays were performed and the results showed that these mutants contain no PEP synthase activity. By using bacteriophage P22, the pps mutations isolated in this study were found to be contransducible with genetic markers in both the aroD and btuC genes. Three factor crosses pinpointed the order of these genes and their distances with respect to each other. One of the mutants carries a pps::lac operon fusion. This fusion was used to explore the transcriptional regulation of the pps gene. A functional copy of the pps gene is required for its own induction. The pps gene is also under catabolite repression, but the addition of adenosine 3',5'-cyclic monophosphate (cyclic AMP) to cells grown in the presence of glucose does not relieve this repression. These results indicate that the synthesis of PEP synthase is regulated in a more complex manner than has been previously recognized. PMID- 2558625 TI - The influence of cold or isolation stress on neuroinvasiveness and virulence of an attenuated variant of West Nile virus. AB - The effect of cold or isolation stress on neuroinvasiveness and virulence was investigated in mice inoculated with an attenuated WNV (WN-25) strain. The WN-25 variant differed from the parent strain by its inability to kill mice after I.P. injection though it was able to immunize even after injection with low doses of virus. Exposure of inoculated mice for 5 minutes a day to cold water (1 +/- 0.5 degrees C) for 8 days resulted in 60% mortality, while in nonstressed infected mice no death was observed. Cold or isolation stress increased the virus level in the brain to 8.9 and 7.4 log 10 PFU as compared to no virus in the infected control. Moreover, it was found that virus level in the spleen of stressed mice reached 3.4 and 3.7 log 10 PFU respectively, while in non-stressed mice no virus was detected. The virus which was isolated from the brain of moribund stressed mice was extremely virulent: I.P. inoculation of as little as 10 PFU caused death to normal non-stressed mice. We suggest that cold or isolation stress conditions in mice inoculated with an attenuated strain induce a selection process. The virus which was isolated from the brain of stressed mice changes its virulence and kills like wild type WNV. PMID- 2558626 TI - Structural analysis of proviral DNA in simian foamy virus (LK-3)-infected cells. AB - Proviral DNA of the T-cell lymphotropic simian foamy virus strain LK-3 was characterized. In infected cells, multiple copies of unintegrated linear duplex viral DNA of about 13 kbp length are present. Nuclease S1 treatment of the DNA generated two fragments of 6.5 and 6.0 kbp length that were cloned in phage and plasmid vectors. The proviral DNA contains a single-stranded gap of 109 nucleotides. A DNA fragment spanning the gap was cloned after completing the double strand by DNA synthesis in vitro. At the 3' end, the gap contains a polypurine tract (PPT) similar to the putative initiation site of retroviral plus strand DNA synthesis, suggesting discontinuous DNA synthesis. Further analysis of the genome architecture revealed LTRs of 1.7 kbp length. An additional 1.7 kbp DNA fragment was detected after nuclease S1 digestion of proviral DNA and probably represents trimmed intermediates of "strong-stop" DNA. PMID- 2558628 TI - A new subgroup 2 bovine adenovirus proposed as the prototype strain 10. AB - A slowly growing subgroup 2 bovine adenovirus (BAV) strain designated Ruakura 78 5371 was isolated from a yearling heifer with systemic adenovirus infection. Cross neutralization tests and restriction endonuclease analysis of the viral DNA showed the virus to be distinct from the other 9 recognised types of BAV. It is proposed that this strain should be regarded as the prototype strain of the new type BAV-10. PMID- 2558627 TI - Unusual human rotavirus strains having subgroup I specificity and "long" RNA electropherotype. AB - During an epidemiological study of human rotavirus in Metro Manila, Philippines, we found 20 unusual strains which belong to subgroup I but have "long" RNA pattern typical of subgroup II human rotavirus. The RNA patterns of the 20 strains were classified into four groups though they were very similar to each other. Four strains, designated L4, L26, L27, and L34 were isolated in MA104 cells from stool specimens. They possessed subgroup I specificity and long RNA pattern identical to that of the viruses in the original stool samples. The serotype specificity of these strains could not be determined by either enzyme linked immunosorbent assay (ELISA) or neutralization test, while one strain (L27) was neutralized by serotype 2-specific anti-VP4 monoclonal antibody. These strains were suggested to have an unusual antigenicity on VP7. PMID- 2558629 TI - Comparison of genomes of malignant catarrhal fever-associated herpesviruses by restriction endonuclease analysis. AB - The restriction endonuclease DNA cleavage patterns of eight isolates of malignant catarrhal fever-associated herpesviruses were examined using the restriction endonucleases HindIII and EcoRI. The eight viruses could be assigned to two distinct groups. Virus isolates from a blue wildebeest, a sika deer and an ibex had restriction endonuclease DNA cleavage patterns that were in general similar to each other. The restriction pattern of these three viruses was distinct from the other five. Of these five, four were isolated from a greater kudu, a white tailed wildebeest, a white bearded wildebeest, and a cape hartebeest. The fifth isolate C500, was isolated from a domestic cow with malignant catarrhal fever. These five viruses had similar DNA cleavage patterns. PMID- 2558630 TI - Pathogenic properties of encephalomyocarditis virus isolates in swine fetuses. AB - The pathogenicity of 5 different encephalomyocarditis (EMC) virus isolates was investigated in swine fetuses following injection of each virus in utero. Laparotomies were performed on 3 pregnant sows in early mid-third (39-40 days) of gestation and on 5 sows in the late mid-third (70-72 days) of gestation, and groups of fetuses were inoculated with different viruses into the amniotic sacs. The uninoculated fetuses served as controls. Thirty-five (71.4%) of 49 infected and 1 of 26 control fetuses were grossly abnormal. Virus was recovered from 18 of 28 infected fetuses and 1 of 16 control fetuses examined. Antibody to EMC virus was detected in all of 14 fetuses infected at 70-72 days of gestation and examined 11-26 days post-infection. The fetal pathogenicity was different depending on the virus strains and the fetal age at the time of virus infection. The EMC ATCC-VR 129 virus was not pathogenic but NVSL-PR, MN-25 and MN-30 were highly pathogenic to the fetuses in both early and late mid-thirds of gestation, while NVSL-MDV was pathogenic to the fetus in early but not in late mid-third of gestation. Possible mechanisms for differences in the pathogenicity between the virus strains are discussed. PMID- 2558631 TI - Drug resistant rhinoviruses from the nose of experimentally treated volunteers. AB - Viruses were isolated from nasal washings of volunteers receiving experimental therapy for rhinovirus type 9 infection with intranasal sprays of a new synthetic antiviral R61837. On a screening test nine subjects yielded drug sensitive virus and four resistant virus. In four others the virus was sensitive at first but became resistant later, while in one the reverse occurred. Evidence is given that at least some of the resistant viruses were present in the respiratory tract and were not selected during virus isolation. Of six viruses studied in detail, five had a low degree of resistance and one was highly resistant. The degree of resistance of the five was similar for an antiviral chalcone, dichloroflavan and disoxaril. The sixth was different in that the resistance to disoxaril was relatively less than to the other drugs. The significance of these results is discussed--these are the first experiments in man to show the selection of drug resistant rhinovirus. PMID- 2558632 TI - Simian alphaherpesviruses and their relation to the human herpes simplex viruses. AB - Biochemical and immunological properties of structural and non-structural polypeptides of the human simplex viruses (HSV1 and HSV2) and four related herpesviruses of non-human primates [Herpesvirus simiae (B virus), H. cercopithicus (SA8), H. saimiri 1 (HVS 1), and H. ateles 1 (HVA 1)] were compared. Using a radioimmunoassay (RIA), the presence of antigenic determinants shared among all six viruses was demonstrated. The relative degree of antigenic cross-reactivity among these viruses was further assessed by competition RIA. Antigenically, HSV 1 and HSV 2 were most closely related to each other although both SA 8 and B virus were also very closely related to HSV 1. Considerably less cross-reactivity existed between either HVS 1 or HVA 1 and the other four primate herpesviruses. Cross-hybridization between simian and human herpesvirus genomes demonstrated that extensive homology exists between each of the simian viruses and both HSV1 and HSV 2. Viral polypeptides bearing common antigenic determinants were identified by immune precipitation of infected cell polypeptides and by immunoblotting. Among the polypeptides of HSV which were recognized by antisera to simian viruses were the VP 5 and p40 proteins, both of which are structural components of the virion nucleocapsid. Using recombinant plasmids containing sequences of the HSV 1 VP5, p40, DNA polymerase, major DNA binding protein, and TK enzyme genes, homologous sequences were detected in all four simian viruses. Together, these results demonstrate that HSV 1, HSV 2, SA 8, and B virus form a closely related sub-group of the primate herpesviruses; HVS 1 and HVA 1 are also related to the other four primate herpesviruses, albeit more distantly. PMID- 2558633 TI - Comparative sequence analysis of VP7 genes from five Australian porcine rotaviruses. AB - The genes coding for the rotavirus major neutralizing protein, VP7, from 5 Australian porcine rotaviruses representing glycoprotein (i.e. VP7 or G) serotypes 3, 4, and 5, were sequenced. The genes were each 1,062 nucleotides long with two long open reading frames for proteins of either 326 or 297 amino acids and containing only one potential glycosylation site at amino acid position 69. When compared to the corresponding genes of human viruses, the porcine genes showed very high nucleotide and deduced amino acid homology. Sequence comparison also revealed that Australian porcine rotaviruses of G serotype 4 and 5 were similar to the corresponding porcine strains found in the U.S.A. and U.K., while G serotype 3 and 4 porcine rotaviruses were closely related to human G serotype 3 strain, RV-3 and serotype 4 strain, ST-3, respectively. These Australian rotavirus VP7 sequences were found to correlate with serological data we reported previously. PMID- 2558635 TI - Studies on attenuation of rotavirus. A comparison in piglets between virulent virus and its attenuated derivative. AB - The development of rotavirus vaccines against acute gastroenteritis for human infants has been accorded a very high priority. Several vaccine candidates all of which are live cultivated strains of animal origin have been tested in humans. However the nature of attenuation of these viruses for humans is unknown. In this study we have attenuated a pig rotavirus by 15 sequential passages in cell culture after which the virus no longer causes diarrhoea in piglets. The pathogenesis of infection of the attenuated rotavirus strain (AT/76 P15) in gnotobiotic piglets was compared with that of the virulent parent strain (AT/76). The pattern of virus replication in the small intestine was judged by histology, disaccharidase assay, immunoperoxidase labelling of gut sections using group A specific rotavirus antibody, and rotavirus antigen assay of gut contents. The parent strain caused variable but extensive infection that resulted in the complete destruction of mature small intestinal enterocytes and villous contraction within 3 days. Membrane bound digestive enzymes were lost, and profound watery diarrhoea and dehydration resulted in causing piglets to become moribund. In contrast attenuated virus appeared to propagate at a much slower pace. Fewer infected epithelial cells were detected at any one time. Destruction of enterocytes was never extensive enough to cause marked mucosal changes in histology. Membrane bound digestive enzymes remained near normal levels and there was little or no diarrhoea. Virus replication ceased after 6 days. It is concluded that attenuation of the porcine rotavirus strain studied was associated with its decreased ability to propagate in enterocytes after adaption to culture. PMID- 2558634 TI - Characterization of a temperature sensitive feline infectious peritonitis coronavirus. AB - The characteristics of a temperature sensitive feline infectious peritonitis virus (TS-FIPV) were examined. TS-FIPV, unlike its parent strain, DF2 wild type FIPV (WT-FIPV), propagated at 31 degrees C (permissive temperature) but not at 39 degrees C (nonpermissive temperature). This temperature preference of TS-FIPV was also demonstrated in cats by the ability of the virus to replicate only at the lower temperature in the upper respiratory tract and not at systemic sites where higher temperatures (38-39 degrees C) prevail. Viral structural proteins and RNA were synthesized at 39 degrees C but some undefined maturational defect prevented the formation of infectious TS-FIPV at its nonpermissive temperature. TS-FIPV was more thermolabile than WT-FIPV which indicated alterations in the structural proteins of TS-FIPV, and a difference in the envelope protein of the two viruses was revealed by Western blot analysis. Plaque assay characterization showed that TS-FIPV produced small plaques in comparison to the large plaques of WT-FIPV. These unique characteristics possessed by TS-FIPV may account for its nonvirulent nature and ability to stimulate protective immune responses in cats. PMID- 2558636 TI - HM-PAO-imaging and herpes encephalitis. AB - Selective uptake of the cerebral blood-flow imaging agent 99mTc hexamethylpropyleneamine oxime (HM-PAO) by Human Herpesvirus 1 (HSV-1) infected cells was investigated in vivo and in vitro. No specific uptake of HM-PAO was observed either in encephalitic rats (by brain scintigraphic imaging or by immunoperoxidase staining/autoradiography of brain sections) or in HSV-1 infected Vero cells. PMID- 2558637 TI - Cystein 402 of HIV gp 120 is essential for CD4-binding and resistance of gp 120 to intracellular degradation. AB - A DNA fragment encoding the CD4-binding region of human immunodeficiency virus type 1 (HIV) gp 120 was excised from an SV40-based expression vector containing gp 160, and subcloned into phage M13 for site-directed mutagenesis. Mutant vectors were constructed and CV-1 cells were transfected with constructs, where Cys402 was substituted for a serine, and metabolically labelled with [3H]-N acetylglucosamine (GlcN). Radioimmunoprecipitation with an hyperimmunserum, specific for gp 120/gp 160, and subsequent SDS-polyacrylamide gel electrophoresis demonstrated presence of gp 160, whereas gp 120 was replaced by [3H]-GlcN labelled material, migrating as a diffuse band corresponding to 80-105k, suggesting increased sensitivity of mutant env gene products to proteolysis after cleavage to gp 120. Wild type gp 120 and gp 160 bound to CD4, whereas neither gp 160 nor gp 120 from mutant-transfected cell lysates did bind to CD4. Altogether the results indicated that Cys402, probably by participating in a disulfide bridge, is essential for (i) the CD4-binding ability of env gene products and for (ii) the physical stability of gp 120. PMID- 2558638 TI - [Malignant fibrous histiocytoma of the neck with metastases to the brain]. AB - A case of malignant fibrous histiocytoma with multiple brain metastases in a 48 year-old man is described. The tumour (5 X 6 cm in size) located under m. sternocleidomastoideus, surrounded inner carotid artery and grew into its wall. Pseudoaneurysm (4 X 5 cm in size) with thrombosis was found 2 cm over the tumour. There were multiple metastases (from 1 to 3 cm in diameter) in the right brain hemisphere. The tumour consisted of fibroblast-like cells, polymorphic histiocyte like cells, with one or several nuclei, Touton cells, and osteoclast-like cells and groups of xanthoma cells. PMID- 2558639 TI - Measurements of probing velocity with an automated periodontal probe and the relationship with experimental periodontitis in the Cynomolgus monkey (Macaca fascicularis). AB - The Toronto Automated Periodontal Probe allows simultaneous measurements of gingival attachment level and probing velocity. The efficacy of probing velocity in measuring the integrity of the dento-gingival junction was studied and compared with loss of gingival attachment level, gingival inflammation, tooth mobility and gingival crevicular fluid collagenolytic activity. The gingiva around the maxillary and mandibular incisor teeth of 2 monkeys were first brought to clinical health by daily prophylaxis. Baseline measurements were made and inflammation was then induced around the maxillary incisors by withdrawing plaque control and by placing silk ligatures subgingivally. The mandibular incisors served as controls. After 10 weeks, the ligatures were removed and the teeth scaled and polished daily for 1 week. The plaque index and gingival index reached maximum values within 1 week; crevicular fluid flow was significantly increased at 1 and 3 weeks but mobility did not change significantly. There was progressive loss of the gingival attachment level, which peaked at 10 weeks on experimental teeth; there was no significant loss on control teeth. Probing velocity around experimental teeth exhibited large and significant increases above controls between 1 and 3 weeks, coinciding with the largest weekly losses of the gingival attachment level. Thus probing velocity may be a useful clinical method for the ancillary measurement of early loss of the gingival attachment level. It can also detect biophysical changes in the dento-gingival junction of progressive inflammatory lesions. PMID- 2558640 TI - Characteristics of collagenase in experimental periapical granulomas from the teeth of dogs. AB - Periapical granulomas were induced, with a success rate of about 60% (66 granulomas produced out of 109 roots treated) in mandibular premolars. The average wet weight of the granulomas, 46.1 +/- 34.5 mg (mean +/- SD, n = 22), was sufficient to allow individual specimens to be used for most of the biochemical analyses. High collagenase activity was extracted directly from the granulomas with 4 M urea solution. The enzyme was a typical animal collagenase (EC 3.4.24.7) which clove native collagen molecules into three-quarter (alpha A) and one quarter (alpha B) length fragments. The collagenase was activated by 1 mM p aminophenylmercuric acetate. This activated enzyme broke down collagen I rather than collagen III preferentially, which is similar to the activity of human polymorphonuclear leucocyte collagenase. The molecular weights of the latent and activated collagenases were 67 and 49 K, respectively. PMID- 2558641 TI - The hydroxyapatite solubility product of human dental enamel as a function of pH in the range 4.6-7.6 at 20 degrees C. AB - Samples of 200 mg powdered enamel apatite were suspended in 10 ml of aqueous solutions made from phosphoric acid and calcium hydroxide for 60 days at 20 degrees C. Calcium and phosphate concentrations were determined by spectroscopy; pH and the concentrations of fluoride and carbonate by electrometry. The solubility product of hydroxyapatite varied linearly from 10-56.9 at pH 4.6 to 10 52.8 at pH 7.6. The carbonate concentration in the aqueous phase was too low to account for calcium carbonate complex formation of significance to the solubility product. After the equilibration, X-ray diffraction analysis of the powder showed that the salt was unchanged pure apatite with no trace of a transformation to brushite or any other calcium phosphate. A re-equilibration of some of the powder samples for another 60 days produced similar solubility products. Thus the change of the solubility product was most likely attributable to a pH-induced change of the ionic composition of the enamel crystal surface. PMID- 2558642 TI - Solubility, unit cell dimensions and crystallinity of fluoridated human dental enamel. AB - The aim was to describe the dimensions of the fluorhydroxyapatite unit cell and the solubility of fluorhydroxyapatite as a function of the fluoride concentration in the apatitic lattice. Various amounts of powdered human dental enamel were suspended in a pH 4.5 acetate buffer containing fluoride. The suspensions were equilibrated for 365 days at 20 degrees C. After equilibration the fluoride concentration in the apatite varied from 0.003 to 0.89 (mole fraction). X-ray diffraction showed that the length of the alpha-axis decreased with increasing fluoride concentration, whilst a broadening of the (300) reflection indicated the presence of mixtures of various fluorhydroxyapatites. The solubility of the apatite after the long equilibration decreased slightly with increasing fluoride concentration in the solid. In a few samples the fluoride concentration in the aqueous phase was still discernible, indicating that even after 365 days, equilibrium had not been attained. In these samples the amount of dissolved apatite was extraordinarily low. PMID- 2558645 TI - Formation of chloramine derivatives of histamine: role of histamine chloramines in bronchoconstriction. AB - Hypochlorous acid generated by myeloperoxidase reacts with histamine to produce chloramines. At pH 7, one mole of histamine monochloramine (HisCl) was generated per mole of H2O2 provided as substrate for myeloperoxidase. At pH 5, one mole of histamine dichloramine (HisCl2) was generated per two moles of H2O2. HisCl and HisCl2 had two and four oxidizing equivalents per molecule, respectively. In vitro, 30 microM HisCl and HisCl2 induced mepyramine-sensitive guinea pig lung parenchyma contraction with 89 and 56 percent of the response of an equivalent concentration of histamine. Pretreatment of lung strips with chloramines reduced the subsequent contractile response of the tissues to methacholine. These results suggest that H-1 histamine receptors provide for targeting of histamine chloramines to pulmonary tissue which may facilitate modification of tissue responses. PMID- 2558643 TI - Radiographic and histological analysis of tooth eruption through calcium phosphate ceramics in the cat. AB - The effect of implanting calcium phosphate ceramics (CPC) into metabolically active sites within kitten mandibles during permanent premolar tooth eruption was examined. Forty kittens, 3-4 months of age were used: the deciduous second and third mandibular premolars were extracted and their sockets implanted with autologous blood clot, autogenous cancellous marrow, and the calcium phosphate ceramics, non-porous beta-tricalcium phosphate or porous hydroxylapatite. Animals were killed at 1, 2, 3, 4 and 5 months after implantation and undermineralized sagittal sections were evaluated by light microscopy. Eighty percent of hydroxylapatite implanted mandibles showed delay in tooth eruption concurrent with distortion in crown development, and a dense cellular fibro-proliferative response within the follicle of unerupted teeth. This response occurred in only one specimen with tricalcium phosphate, whereas normal eruptive patterns and crown development were routinely noted. Both the tricalcium phosphate and hydroxylapatite were integrated into the surrounding alveolar bone without evidence of an inflammatory response. Thus hydroxylapatite initiated a dense cellular fibrous network within the dental follicle preventing formation of an eruptive pathway, delaying tooth eruption and causing crown deformation. This was rarely seen with tricalcium phosphate, and may be due to the resorbability of tricalcium phosphate when compared to hydroxylapatite. Hydroxylapatite should therefore be used with caution for implanting into areas containing unerupted teeth with a metabolically active dental follicle. PMID- 2558646 TI - The role of the carboxyl-terminal 6 amino acid extension of human TSH beta subunit. AB - The nucleotide sequence of human thyroid stimulating hormone (hTSH) gene can encode a protein of 138 amino acids. However, the mature polypeptide is lacking 6 amino acids of the carboxyl-terminus (C-terminus), suggesting posttranslational cleavage of these residues. To analyze a possible function of these 6 amino acids, we expressed two hTSH beta cDNAs with or without the 6 codons for C terminal extension, together with alpha subunit cDNA in CHO cells, and determined the amino acid sequence of C-terminus of hTSH beta. hTSH beta propeptides without C-terminal extension were glycosylated, associated with alpha subunit and secreted, as normal propeptides were, and its heterodimer with alpha subunit showed normal TSH bioactivity in FRTL-5 bioassay. These data indicate that the 6 amino acid C-terminal extension is not necessary for the hTSH maturation in the process of the biosynthesis and for its bioactivity. PMID- 2558644 TI - Detection of hydroxyl radical in the mitochondria of ischemic-reperfused myocardium by trapping with salicylate. AB - Although the presence of free radicals has been indicated in ischemic-reperfused heart, the exact nature and source of these free radicals are not known. The present study utilized a chemical trap, salicylic acid, to trap hydroxyl radical which could be detected as hydroxylated benzoic acid using high pressure liquid chromatography. Since the hydroxylated product is extremely stable, heart was subjected to subcellular fractionation after ischemia and reperfusion, and each fraction was separately examined for the presence of hydroxyl radical. The results indicated for the first time the presence of hydroxyl radical in the mitochondrial fraction during early reperfusion, which decreased in intensity as the reperfusion progressed. PMID- 2558647 TI - Endotoxin increases the liver fructose 2,6-bisphosphate concentration in fasted rats. AB - Following endotoxin administration to fasted rats, the liver fructose 2,6 bisphosphate level is significantly increased within 1 hr, is elevated 2.3-fold by 3 hrs, and remains elevated 2 to 3-fold for at least 24 hrs. This increase in the potent allosteric activator of phosphofructokinase occurs when there is no change in the liver Glc 6-P, glycogen or cAMP concentrations, or in the activities of phosphoenolpyruvate carboxykinase or pyruvate kinase. The increase in fructose 2,6-bisphosphate concentration accounts for the increased phosphofructokinase activity previously observed in hepatocytes isolated 18 hours following endotoxin administration to rats (1). By stimulating the phosphofructokinase/Fru 1,6-bisphosphate cycle in the direction of glycolysis, fructose 2,6-bisphosphate is likely the factor responsible for decreased gluconeogenesis in endotoxemia. PMID- 2558648 TI - Effects of recombinant interleukin-1 beta on phospholipase A2 activity, phospholipase A2 mRNA levels, and eicosanoid formation in rabbit chondrocytes. AB - We have investigated the effects of recombinant interleukin-1 beta (rIL-1 beta) on phospholipase A2 activity (PLA2), PLA2 messenger RNA levels, and eicosanoid production in rabbit chondrocytes. Phospholipase A2 activity increased 5 fold with exposure to 1.6 x 10(-11) M rIL-1 beta for 20 hours. An mRNA specific for an extracellular PLA2 was detected by RNA blot hybridization after treatment of the cells with rIL-1 beta. Hydroxylated derivatives of arachidonic acid, including prostaglandins, leukotriene B4, 5-hydroxyeicosa 6E, 8Z, 11Z, 14Z-tetraenoic acid, 12-hydroxyeicosa 5Z, 8Z, 10E, 14Z-tetraenoic acid and 15-hydroxyeicosa 5Z, 8Z, 11Z, 13E-tetraenoic acid increased in cells after rIL-1 beta treatment. PMID- 2558649 TI - A novel a-type terminal oxidase from Sulfolobus acidocaldarius with cytochrome c oxidase activity. AB - Cytochrome C oxidase was solubilized with a nonionic detergent n-decanoyl-N methyl glucamide from the membranes of Sulfolobus acidocaldarius, a thermoacidophilic archaebacterium, and was purified. The enzyme oxidized horse heart cytochrome C with a Vmax of 63 mumols/min/mg at 50 degrees C. The activity was sensitive to cyanide. The enzyme also catalyzed oxygen uptake detergent on N, N, N', N'-tetramethyl p-phenylene diamine. An apparent molecular mass was estimated to be 150 kDa. The enzyme is composed of three subunits of 37, 23 and 14 kDa. Spectral characteristics were similar to typical bacterial aa3 except for the presence of a novel 583 nm peak observed in reduced minus oxidized difference spectrum. PMID- 2558651 TI - Molecular cloning, sequence and functional expression of the cDNA for the human thyrotropin receptor. AB - We screened a human thyroid cDNA library with two synthetic oligonucleotides based on the reported amino acid sequence of the 3rd and 4th transmembrane domains of a putative human TSH receptor and related receptors. The nucleotide sequence of a 4 kb clone revealed an open reading frame of 764 amino acids (86,816 Daltons) with a putative signal peptide, seven transmembrane domains, five potential glycosylation sites, and a very short intracytoplasmic region. Homology with the extracellular domain of the pig LH/CG receptor was only 33%. Chinese hamster ovary cells stably transfected with this cDNA in an expression vector generated a functional receptor, able to activate adenylate cyclase, specifically in response to TSH stimulation. PMID- 2558650 TI - c-myc gene expression in human cells is controlled by glucose. AB - The c-myc oncogene is implicated in normal growth and differentiation processes. Human cell lines IM9 and HepG2 stably cultured at "low" glucose concentrations (5.5 mM) show c-myc mRNA levels 3-4 times higher than cells cultured at "high" glucose concentrations (25 nM). D-fructose (a metabolizable exose) substitutes for D-glucose in reducing c-myc expression while 3-ortho-methylglucose (a non metabolizable exose) is uneffective. c-myc expression is up-regulated (by PMA) or down-regulated (by dexamethasone and long-term exposure to FCS) in human cells cultured at "low" glucose but not in cells cultured at "high" glucose. We previously demonstrated that insulin receptor gene expression in human cell lines in enhanced by glucose. Therefore, glucose controls in an opposite way the expression of two genes important in the regulation of eukaryotic cell growth and differentiation. PMID- 2558652 TI - Hypericin and pseudohypericin specifically inhibit protein kinase C: possible relation to their antiretroviral activity. AB - Hypericin and pseudohypericin which have been isolated from plants of the Hypericum family are aromatic polycyclic diones. Daniel Meruelo et. al. have reported that hypericin and pseudohypericin showed potent antiretroviral activity including anti-human immunodeficiency virus (1,2). However, the mechanism of these antiretroviral activities has not been clarified. In the course of screening specific inhibitors of protein kinase C we have found that both compounds specifically inhibit protein kinase C with IC50 values 1.7 micrograms/ml and 15 micrograms/ml, respectively, and show antiproliferative activity against mammalian cells. These data suggest that antiretroviral activity of hypericin and pseudohypericin could be attributable to the inhibition of some phosphorylation involved by protein kinase C during viral infection of cells. PMID- 2558654 TI - The second messenger system as the morphogenetic field. AB - For sixty years morphogenetic fields have been assumed. We suggest a realization for such a field: the second messenger (in particular cyclic AMP) and its substrate (in particular ATP) are suggested to be the activator-morphogen and inhibitor respectively. Gene transcription is derived as an hysteretic function of the second messenger (cyclic AMP). PMID- 2558653 TI - Structure of mouse 2',3'-cyclic-nucleotide 3'-phosphodiesterase gene. AB - The mouse 2',3'-cyclic-nucleotide 3'-phosphodiesterase gene was isolated from a mouse gene library. Restriction endonuclease mapping and DNA sequencing analysis revealed that this gene is about 6 kb long and is separated into three exons by two introns. The transcription initiation site was identified. The mouse cDNA of 2374 bp was obtained and used for the screening and analysis of the gene. PMID- 2558655 TI - Selective inhibition of tyrosine protein kinase by a synthetic multisubstrate analog. AB - Synthetic compounds were designed in an attempt to mimic the possible transition state of tyrosine protein kinases. One representative compound (RP 53801) inhibited the enzyme purified from RSV-transformed cells. A serine/threonine kinase (kinase C) was 45 fold less sensitive. The inhibition was competitive with respect to ATP and noncompetitive with respect to the phosphate acceptor poly glu4-tyr1. The degree of inhibition (IC50 = 22 microM) was however lower than that expected from a transition state analog. The compound was capable of reducing tyrosine protein kinase activity in intact cells with some selectivity at 100 microM. PMID- 2558656 TI - Sequence of rat skeletal muscle phosphoglycerate mutase cDNA. AB - A cDNA clone coding rat skeletal muscle phosphoglycerate mutase was isolated from a rat muscle lambda gt10 cDNA library and its sequence was determined. The deduced protein possesses 252 amino acids and is 94% homologous with respect to human muscle phosphoglycerate mutase. No amino acids changes occur at the active site and structural predictions suggest strong conformational homologies with other enzymes of the mutase family. PMID- 2558658 TI - Heterogeneity of rat ovarian lactogen receptor species. AB - Three distinct ovarian lactogen receptor species with unique and highly reproducible HPLC retention times gave corresponding peaks of binding activity and migrated as single bands of 80, 40, and 34 kDa on SDS-PAGE. Reduction of the 80 kDa protein failed to reveal any conversion to the lower molecular weight proteins by either SDS-PAGE analysis or reverse phase HPLC, suggesting that the three binding proteins are not related by disulfide bond formation. Immunological studies indicate an amino acid homology at a C terminal region among the 3 receptor forms and with the rat liver receptor. However, the 80 kDa ovarian receptor also contains a unique sequence derived from microsequencing that is not immunologically apparent in the ovarian lower molecular weight forms of the rat liver receptor. These findings substantiate the existence of at least two populations of ovarian lactogen receptors perhaps originating within the same gene, a high Mr form potentially capable of signal transduction, and truncated forms that could be involved in transport and/or clearance. PMID- 2558657 TI - Evidence for a second isoform of the catalytic subunit of calmodulin-dependent protein phosphatase (calcineurin A). AB - We have used a previously characterized rat cDNA clone for the catalytic (A) subunit of calmodulin-dependent protein phosphatase (calcineurin), which we designated A alpha, to isolate cDNA clones coding for a second isoform of the A subunit, A beta. The A beta cDNA encodes a protein of 525 amino acids that is 81% identical with A alpha. The N-terminal region is dissimilar and contains a characteristic proline-rich sequence. The region homologous to protein phosphatases 1 and 2A (region between residues 87 and 338, 91% identical) and the calmodulin binding domain (region between residues 401 and 424, 96% identical) are highly conserved. The presence of two genes coding for calcineurin A suggests the possibility of important functional differences in the two enzymes. PMID- 2558659 TI - Fibronectin modulates the activation of human platelets. AB - In this paper we studied the effect of plasma fibronectin on platelet responsiveness to low doses of thrombin or ADP. Fibronectin causes: (i) a significant lowered cytoplasmic calcium movement in platelets activated both with low doses of thrombin and with ADP, (ii) a lowered decrease of the cAMP level induced by low thrombin, but not by ADP, (iii) a dramatic decrease of protein phosphorylation in low thrombin-treated platelets. The results reported here demonstrate that plasma fibronectin behaves like an inhibitor of mild activations of human platelets; its effect is probably mediated by the glycoprotein llb-llla complex in thrombin activated platelets, whereas a different and unknown mechanism must be supposed to explain the results obtained with ADP-activated platelets. PMID- 2558660 TI - Influences of tromantadine and nonoxinol 9 on the stability of red cell membrane. AB - The antiviral compound tromantadine (ViruMerz) and the detergent nonoxinol 9 have been investigated in their effects on biophysical parameters of red cell membranes. Up to a maximum ratio of 1 mol per 800 mol of phospholipids tromantadine enhances the membrane phase transition/separation break at 16-20 degrees C measured by 1-anilinonaphthalene-sulfonate (ANS) fluorescence. It also increases order parameters obtained from spin labeling experiments with 5-doxyl stearic acid. Nonoxinol 9 interacts with the membrane at a maximum ratio of 1 mol per 40 mol of phospholipids determined by UV spectrophotometry. The substance decreases the intensity of the above phase transition/separation break and the order parameters of the spin label 5-doxyl stearic acid. These experiments indicate that tromantadine probably stabilizes the membrane whereas nonoxynol 9 exhibits opposite effects. Combination of both compounds equalizes the influences of the single substances on the above biophysical parameters of red cell membrane with predominance of the nonoxinol 9 effect. PMID- 2558662 TI - Effect of lysophosphatidylcholine on atherosclerotic rabbit arteries. AB - We report here on the effect of an endothelium-dependent vascular smooth muscle relaxant, lysophosphatidylcholine (LPC) on rabbit aortic strips and on hemodynamic changes by LPC in atherosclerotic animals. Cyclic GMP changes induced by LPC in atherosclerotic vessels were also determined. Atherosclerosis was produced by feeding a high cholesterol and saturated fatty acid diet. LPC was injected into the left atrium and coronary flow was measured by radioactive microspheres; in vitro, relaxation of precontracted aortic strips by lysophosphatidylcholine was also recorded. LPC failed to increase coronary flow in the presence of atherosclerosis. In isolated aortic strips, dose-response curves with acetylcholine and LPC showed diminished relaxation in atherosclerotic preparations, and cyclic GMP production following LPC was reduced. The results demonstrate that vascular relaxation by LPC, together with its ability to activate guanylate cyclase is dependent on the functional and morphological integrity of the vascular wall. PMID- 2558661 TI - Antiproliferative action of cyclic GMP-elevating vasodilators in cultured rabbit aortic smooth muscle cells. AB - In cultured rabbit aortic smooth muscle cells (SMCs), sodium nitroprusside (SNP) (10(-7) to 10(-4) M), atrial natriuretic peptide (ANP) (10(-9) to 10(-6) M) and 8 bromo-cyclic GMP (10(-6) to 10(-3) M) inhibited the whole blood serum (WBS) induced DNA synthesis by about 30%. The doses of SNP and ANP necessary for the inhibition of the WBS-induced DNA synthesis were similar to those necessary for the formation of cellular cyclic GMP (cGMP). These agents were effective even when added 6 h after stimulation of the cells with WBS. These results suggest that cGMP inhibits the proliferation of rabbit aortic SMCs by inhibiting the progression from the G1 into S phase of the cell cycle and raise the possibility that cGMP-elevating vasodilators may suppress the atherogenic process by inhibiting vascular SMC proliferation. PMID- 2558664 TI - [Association of an adenoid cystic epithelioma with a clear cell carcinoma of the palate]. AB - An infrequent polymorphous minor salivary gland tumor is described. It involved the palate and later invaded the upper jaw. The morphologic features are those of both adenoid cystic carcinoma and clear cell carcinoma. The clinical course, the cellular admixture of tumour components, and the immunohistologic results allow us to propose that clear cell carcinoma originating from adenoid cystic carcinoma offers the worst prognosis. PMID- 2558663 TI - [Cell markers, hormone receptors and proliferation index in breast cancers. Role of immunodetection, image analysis and flow cytometry]. PMID- 2558665 TI - [Heart transplant: development and maturity of the program at the University of South Florida and the General Hospital of Tampa]. AB - Cardiac transplantation has evolved from an experimental procedure to an accepted mode of therapy that prolongs life in patients with severe heart failure. The University of South Florida-Tampa General Hospital began performing cardiac transplantation for the treatment of end-stage cardiac disease in June 1985. Since then, 50 heart transplantations have been performed and 37 patients are alive and well. The one-year actuarial survival is 78% and the two-year actuarial survival is 72%. Multiple complications have been encountered, most notably rejection and infection. The recent approval of the USF/TGH program as a Medicare funded cardiac transplantation center is expected to greatly expand the number of potential recipients and will provide the residents of Florida, the Southeastern United States and the Caribbean with an additional health care resource. PMID- 2558666 TI - [Paranasal sinus tumors with orbital involvement]. AB - Paranasal sinus tumors frequently extend into the bony orbital contents. In the past, their treatment often resulted in severe visual defects or consisted in enucleation. In an effort to determine whether an effective but more conservative form of therapy, is possible, 40 patients with such lesions were treated over the past 11 years by microsurgical resection followed by radiotherapy and/or chemotherapy. The present paper describes the authors' surgical technique, i.e. the transfrontal extradural approach and the midface-degloving-technique. In addition, the following parameters are evaluated specifically: 1) the effects of conservative resection on overall survival rate, 2) frequency of local disease recurrence and 3) changes in orbital function as a result of the combined therapy. PMID- 2558667 TI - Anti-idiotypic antibodies as vaccines to a murine Sendai virus infection. PMID- 2558668 TI - Subpopulations of human peripheral T gamma delta lymphocytes. PMID- 2558669 TI - Binding of radiation leukemia viruses to a thymic lymphoma involves some class I molecules on the T cell as well as the T cell receptor complex. AB - Radiation leukemia virus (RadLV)-induced thymomas and malignant thymocytes from AKR mice have been shown to bind specifically retrovirus produced by these cell lines. Each lymphoma has been shown to have greatest specificity for cognate virus suggestive of an immune-specific receptor. The question of receptor identity has been addressed here using the RadLV-induced murine T cell lymphoma, C6VL/1, and antibodies specific for known cell surface determinants present on these cells. This lymphoma has been shown to bind both homologous and heterologous RadLV isolates, but to have greatest specificity for homologous retrovirus since homologous free virions can best block the interaction between cells and virus adhered to the wells of a microtitre plate. A clonotypic anti-TCR antibody has been shown to completely inhibit C6VL/1 binding to the homologous virus, RadLV/C6VL, but not to the heterologous virus, RadLV/VL3. Anti-CD4, anti Thy1.2 as well as anti-H-2Kb and not anti-H-2Db antibodies were found to partially inhibit the interaction with both RadLV/C6VL and RadLV/VL3, yet neither of these virus preparations appears to be contaminated with Class I molecules as measured by radioimmunoassay. The binding interaction between C6VL/1 and RadLV/C6VL appears specifically to involve the TCR since antibody against the clonotypic site on the TCR heterodimer uniquely inhibits this interaction, while the binding of C6VL/1 to RadLV/VL3 appears to involve the H-2Kb molecule. When free virus particles were absorbed to receptors on C6VL/1, both RadLV/VL3 and RadLV/C6VL inhibited the binding of antibody to the TCR and CD4 molecules, while the binding of several anti-H-2Kb antibodies was specifically inhibited by RadLV/VL3. There are at least two known T cell surface structures involved in the interaction of the T cell lymphoma, C6VL/1, with RadLV. These are the TCR complex (comprising the TCR heterodimer and CD4), and the Class I H-2Kb molecule. Since the TCR molecule has been shown to comodulate with H-2Kb molecules when cells were cultured in the presence of anti-H-2Kb antibodies, and the CD4 and H-2Kb molecules have been shown to comodulate with the TCR on only a subpopulation of C6VL/1 cells treated with anti-TCR antibody, this suggests that the H-2Kb molecule may also be part of the larger molecular complex including CD4/8 which can form around the TCR heterodimer.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2558670 TI - [Chemical constituents of Dysosma aurantiocaulis (H.-M.) Hu and Dysosma pleianthum Woods]. AB - Roots of D. aurantiocaulis and D. pleianthum contain ten lignans: picropodophyllin, podophyllotoxin, 4'-demethylpodophyllotoxin, diphyllin, dehydropodophyllotoxin, podophyllotoxone, picropodophyllone, isopicropodophyllone, 4'-demethylpodophyllotoxone and dysosmajol; two anthraquinones: physcion and dysoanthraquinone. A HPLC method for the identification of aryltetralin lignans is described. PMID- 2558671 TI - Ultrasound case study: primary hepatocellular carcinoma. PMID- 2558672 TI - Signal transduction during craniofacial development. PMID- 2558673 TI - The Ah locus: genetic differences in toxicity, cancer, mutation, and birth defects. PMID- 2558674 TI - Opioidlike effects of intraoral infusions of corn oil and polycose on stress reactions in 10-day-old rats. AB - The effects of intraoral infusions of corn oil and the polysaccharide Polycose on behavioral reactions to pain and to social isolation were studied in 10-day-old albino rat pups. Both substances significantly increased paw-lift latencies (a measure of pain response) and reduced the number of ultrasonic vocalizations (a measure of isolation distress). Moreover, elevated pain thresholds were normalized by naltrexone (0.25 mg/kg) pretreatment, and the quieting of vocalizations was abolished by pretreatment. These findings indicate an interaction between ingestion, pain, and distress systems in neonatal rats and suggest that fats and polysaccharides influence these systems via endogenous opioids. PMID- 2558675 TI - Role of ACTH in recovery from retrograde amnesia induced by hypothermia in rats. AB - The present investigation assessed whether increased congruency between ACTH state present shortly after training and that at testing contributed to memory recovery. If recovery were related to an increased correspondence between internal state present after training and that at testing, then suppressing ACTH release should block memory recovery. This was the hypothesis that was examined in the present investigation. Specifically, animals were trained on a passive avoidance task, administered hypothermia (the amnestic agent) and, shortly prior to testing, given treatments known to be effective in reversing memory loss induced by hypothermia. Before training (Experiment 1) or testing (Experiment 2) animals were injected with either dexamethasone (an agent that suppresses ACTH release) or saline. Results, in general, indicated that when ACTH release was suppressed, a blunted recovery effect was obtained. This reduction in the extent of memory recovery was observed when ACTH was suppressed either at training or at testing. These data are interpreted as providing support for an ACTH-related, state-dependent retention mechanism contributing to recovery from hypothermia induced retrograde amnesia in rats. PMID- 2558676 TI - Angiotensin-converting enzyme in subfornical organ mediates captopril-induced drinking. AB - These experiments tested whether angiotensin-converting enzyme (ACE) located within the subfornical organ (SFO) participates in the generation of water intake during peripheral ACE blockade with captopril (CAP). Lesions of the SFO virtually abolished drinking in response to intraperitoneal CAP injection. Intracranially injected CAP suppressed drinking induced by intraperitoneal CAP more completely with direct SFO injection compared with intraventricular or control tissue injections. This central captopril treatment did not alter the drinking response to subcutaneous hypertonic saline. Intraventricular injections of the angiotensin II (ANG II) receptor blocker sarile reduced drinking during oral captopril treatment in rats rehydrating from water deprivation. The results indicate that (a) the SFO mediates drinking caused by peripheral ACE inhibition; (b) the ACE located within the SFO may locally convert ANG I to ANG II, which then stimulates thirst; and (c) central ANG II receptors mediate thirst caused by peripheral ACE inhibition. PMID- 2558678 TI - Ability of periaqueductal gray subdivisions and adjacent loci to elicit analgesia and ability of naloxone to reverse analgesia. AB - This study is an investigation of the effects of stimulation of regions within and adjacent to the periaqueductal gray (PAG) matter. Eighty-five rats were implanted with 1 monopolar stimulating electrode into 1 of 5 loci. Potency of analgesia was evaluated by relative increases in tailflick latencies after brain stimulation, and threshold current intensity was used to elicit analgesia. The ability of naloxone to reverse the stimulation-induced analgesia was also evaluated. Results replicate the previous finding of differential naloxone reversibility of ventral vs. dorsal PAG sites, but they do not support a regional distinction in the potency of analgesia induced. The results suggest that dorsal PAG sites are involved in a separate nonopiate pain-inhibitory system, whereas ventral sites are involved in an opiate system. These systems, however, do not respect the cytoarchitectural boundaries of the PAG because sites adjacent to the PAG elicit similar effects with a corresponding dorsal-ventral distinction. PMID- 2558677 TI - Lesions of the nucleus accumbens in rats reduce opiate reward but do not alter context-specific opiate tolerance. AB - Bilateral electrolytic lesions of the nucleus accumbens in rats eliminated the capacity of 10 mg/kg morphine to produce a conditioned place preference (Experiment 1). However, these lesions did not alter the capacity to establish context-specific tolerance to the analgesic effects of 5 mg/kg of morphine (Experiment 2). This latter finding indicates that rats with nucleus accumbens lesions are not impaired in associating the effects of morphine with a particular location. Thus, the failure of morphine to produce a conditioned place preference in these lesioned rats probably cannot be attributed to an inability to associate the effects of morphine with a particular chamber, i.e., the initially nonpreferred chamber. Rather, morphine may fail to establish a conditioned place preference in these rats because nucleus accumbens lesions disrupt a pathway that is critical in mediating the rewarding effects of opiates. PMID- 2558680 TI - Frontal cortex lesions block the opioid and nonopioid hypoalgesia elicited by brief shocks but not the nonopioid hypoalgesia elicited by long shocks. AB - Previous research (Grau, 1987a, 1987b) suggests that forebrain systems play an essential role in the hypoalgesia observed after brief shock but not long shock. Additional research has shown that pentobarbital anesthesia and decerebration block the hypoalgesia observed after 3 brief (0.75-s) shocks but not the hypoalgesia observed after 3 long (25-s) shocks. This is a study of whether a specific forebrain lesion, a frontal cortex lesion, would have a similar impact on hypoalgesia induced by brief (0.75 s) and long (25-s) shocks. Frontal cortex lesions, like decerebration and pentobarbital anesthesia, eliminated the hypoalgesia observed after brief but not long shocks. Because other research suggests that the stress of surgery may influence whether the hypoalgesia elicited by shock is opioid or nonopioid, the 2nd experiment was to examine whether the sham operation per se alters the form of the hypoalgesia observed after brief shock. It does not; in the sham-treated subjects, brief shock induced the usual transient nonopioid hypoalgesia followed by prolonged opioid hypoalgesia. These data suggest that frontal cortex lesions block nonopioid and opioid hypoalgesia observed after brief shock. PMID- 2558679 TI - Assessment of the neural substrate for intracranial self-stimulation by the postreinforcement pause technique. AB - The poststimulation excitability of neurons mediating intracranial self stimulation (ICSS) was evaluated by the paired-pulse method. Stimulus effectiveness was assessed by the postreinforcement pause (PRP) and by frequency threshold (FT) determinations in 7 rats performing ICSS in the medial forebrain bundle (MFB) and in the ventral tegmental area (VTA). Stimulus effectiveness values were minimal at conditioning-test (C-T) pulse intervals of 0.6 and 0.8 ms for MFB and VTA animals, respectively, because of neuronal refractoriness. Local potential summation could account for the increase in effectiveness at very short C-T intervals, and an additional peak of enhanced effectiveness at a C-T interval of 2.0 ms, perhaps reflecting synaptic events, was observed only in VTA animals with the PRP method. Important advantages of the PRP method were that the C-T interval was the only stimulus parameter that was varied, and the behavioral output of the animal remained relatively constant. PMID- 2558681 TI - Phase-transfer-catalysed synthesis of N-acetylneuraminic acid alpha-thioketosides and inhibitor studies with Clostridium perfringens sialidase. AB - The alpha-thioketosides of methyl 5-acetamido-4,7,8,9-tetra-O-acetylneuraminate with thioacetic acid, thiophenol, 4-nitrothiophenol, 4-aminothiophenol, 2 mercaptopyridin and mercaptobenzothiazol as aglycones were synthesized by phase transfer catalysis in good yields. The methyl 5-acetamido-4,7,8,9-tetra-O-acetyl 2-thioacetylneuraminate is the analogous thio compound to the methyl 5-acetamido 2,4,7,8,9-penta-O-acetylneuraminate and can be used as intermediate for preparing S-ketosides of Neu5Ac. By Zemplen saponification and mild hydrolysis of the methyl-ester group the free Neu5Ac-alpha-thioketosides with thiophenol, 4 nitrothiophenol, 4-aminothiophenol and 2-mercaptopyridin could be prepared. These ketosides were found to be inhibitors of C. perfringens sialidase with Ki-values between 2.3mM and 6.6mM. The free Neu5Ac-alpha-mercaptobenzothiazolyl ketoside could not be prepared by this procedure. It was completely hydrolysed during Zemplen saponification and methyl-ester hydrolysis in alkaline medium. PMID- 2558682 TI - Fc receptors mediate prostaglandin and superoxide synthesis in cultured rat Kupffer cells. AB - Latex beads with covalently bound bovine serum albumin were prepared and coated with anti-BSA immunoglobulin G. These particles were shown to possess on their surfaces a defined quantity of the antibody with the Fc portions exposed to the medium. One homologous and two heterologous antibodies of the G class were used and compared in terms of their binding to the rat Kupffer cells and their ability to elicit the typical phagocytotic responses. These particles were phagocytosed by rat Kupffer cells and elicited synthesis of prostaglandins and superoxide anion radicals. A significant release of superoxide into the medium was observed in the presence of cytochalasin B only. The data presented here suggest that a) Fc-carrying particles can be bound to Kupffer cells and elicit responses via specific receptors; b) coating with the homologous antibody yields the most effective particles; c) superoxide release into the surrounding medium is most abundant when the particle-binding membrane areas are prevented from forming phagocytotic vesicles. PMID- 2558683 TI - Protection of mice from Semliki Forest virus infection by lymphocytes treated with low levels of interferon. AB - These studies provide the first evidence that adoptive transfer of syngeneic mouse (BALB/c) lymphocytes treated with low levels of mouse interferon (IFN) alpha/beta can result in sufficient protection to protect mice from Semliki Forest virus (SFV) infection. Specifically, intraperitoneal inoculation of noncytotoxic lymphocytes treated exogenously with IFN (3 to 50 U/ml), washed exhaustively, and mixed with antibody to IFN-alpha/beta to neutralize any residual or early produced IFN, resulted (after repeated studies) in a 35% to 40% reduction in mortality of mice challenged with SFV (P less than or equal to .01), while inoculation of control lymphocytes had no effect. Direct administration of relatively high levels of IFN-alpha/beta (2,000 U/d) only moderately reduced the mortality (by 20%) in mice. Passive transfer of IFN-treated BALB/c mouse embryo cells also did not protect. The protection could not be attributed to carryover of IFN by the lymphocytes, endogenous IFN induction, enhanced cytotoxicity of endogenous splenocytes or peritoneal leukocytes, or early appearance of antiviral neutralizing antibody. Thus, the most likely cause of the observed protection is consistent with a unique mechanism that can be activated by the IFN-treated lymphocytes. PMID- 2558684 TI - Human trials of AIDS vaccines: current status and future directions. PMID- 2558685 TI - HIV genome variability in vivo. PMID- 2558686 TI - Use of simian immunodeficiency virus for evaluation of AIDS vaccine strategies. PMID- 2558687 TI - Molecular biology of HIV: new insights into the virus life-cycle. PMID- 2558688 TI - Feline immunodeficiency virus is a lentivirus associated with an AIDS-like disease in cats. PMID- 2558689 TI - Structures of three DNA cross-linking agents, ethane-1,2-di(methylsulfonate), propane-1,3-di(methylsulfonate) and n-butane-1,4-di(methylsulfonate). AB - (I): C4H10O6S2, Mr = 218.25, P2(1)/c, a = 7.2611 (9), b = 5.8726 (4), c = 10.6628 (19) A, beta = 103.95 (1) degree, V = 441.3 (2) A3, Z = 2, Dm = 1.65, Dx = 1.642 Mg m-3, lambda(Cu K alpha) = 1.54184 A, mu = 5.43 mm-1, F(000) = 228, R = 0.0336 and wR = 0.0346 for 745 unique reflections with F greater than or equal to 3 sigma (F), T = 298 K. (II): C5H12O6S2, Mr = 232.28, P2(1)/c, a = 11.030 (1), b = 8.452 (1), c = 11.162 (1) A, beta = 104.65 (1) degree, V = 1006.8 (3) A3, Z = 4, Dm 1.54, Dx = 1.532 Mg m-3, lambda(Cu K alpha) = 1.54184 A, mu = 4.79 mm-1, F(000) = 488, R = 0.0418 and wR = 0.0430 for 1666 unique reflections with F greater than or equal to 3 sigma(F), T = 298 K. (III): C6H14O6S2, Mr = 246.30, P1, a = 5.6147 (9), b = 6.8343 (6), c = 7.5434 (6) A, alpha = 110.61 (1), beta = 92.08 (1), gamma = 76.15 (1) degree, V = 262.7 (3) A3, Z = 1, Dm = 1.56, Dx = 1.557 Mg m-3, lambda(Cu K alpha) = 1.54184 A, mu = 4.79 mm-1, F(000) = 130, R = 0.0397 and wR = 0.0425 for 907 unique reflections with F greater than or equal to 3 sigma(F), T = 298 K. All three compounds have a common conformation in the C.O.SO2.CH3 part of the molecules due to weak O...H interactions. Compounds (I) and (III) have a trans conformation about the central C--C bond, whereas (II) is cis. PMID- 2558690 TI - Red blood cell Na+/H+ and Li+/Na+ exchange in patients with essential hypertension. AB - The increased red blood cell Li+/Na+ exchange found in a subgroup of patients with essential hypertension (EH) may reflect an increased activity of the Na+/H+ exchange. The maximal velocity of the red cells' Na+/H+ (Na+ influx promoted by an outward H+ gradient) and Li+/Na+ (Li+ efflux promoted by external Na+) exchange were therefore measured in 41 EH and in 21 normotensive controls (NT). Both transporters were significantly higher in EH than in NT (74 +/- 39 mmol/L cell x h v 43 +/- 27 for the former, P less than .03, and 0.35 +/- 0.16 v 0.26 +/ 0.10 for the latter, P less than .05). Even though more than 100 times faster, Na+/H+ exchange was weakly but significantly correlated to Li+/Na+ exchange (r = 0.29, P less than .05). Proximal tubule Na+ reabsorption (fractional renal Li+ reabsorption) was significantly greater in EH than in NT (0.78 +/- 0.07, n = 32, v 0.73 +/- 0.06, n = 10, P less than .05) but it was not correlated to either the red cells' Na+/H+ or Li+/Na+ exchanges. Therefore, hyperactivity of Na+/H+ exchange in EH may play a role in blood pressure elevation through mechanisms other than stimulation of renal Na+ reabsorption. PMID- 2558691 TI - Low-molecular-weight heparin Fraxiparin in chronic hemodialysis. A dose-finding study. AB - A two-step dose-ranging study was undertaken with CY216 (Fraxiparin) in 8 patients on 7 sessions each. The different doses were administered each time as a single bolus injection at the start of hemodialysis without heparinized priming nor further administration during the 4-hour session. In the first step, the clinical efficacy of 4 different doses of Fraxiparin was compared with that of standard heparin on the percentage of sessions free of clot formation in the extracorporeal circulation (ECC). Safety was evaluated by the compression time for hemostasis at the puncture sites. The second step was a randomized comparison of 3 Fraxiparin dosages. In addition to the clinical assessment of efficacy, the following biological parameters were measured: fibrinopeptide A (FPA), anti-Xa (AXa) activity calibrated against Fraxiparin, thrombin time (TT), activated partial thromboplastin time, blood counts, hemoglobin, hematocrit, plasma creatinine and urea, and residual blood volume in the dialyzer. A 'standard' dosage of 10,000 AXa Institut Choay units of Fraxiparin was shown to prevent clot formation in the ECC. It resulted in a marked increase in TT, without any lengthening of the puncture site compression time. After 4 h, AXa and FPA levels in the venous line were related to the doses used (p less than 0.05). After 48 h AXa activity was very low. Dialysance and tolerance were excellent. Thus, a single dose of Fraxiparin unrelated to body weight and not determined by the measurement of the whole-blood activated clotting time appeared to be a safe and effective means for preventing fibrin formation in 4-hour dialysis sessions. PMID- 2558692 TI - Mitogenic properties of two distinct forms of toxic shock syndrome toxin-1 separated on hydroxyapatite by high-performance liquid chromatography. AB - The homogeneity of a purified staphylococcal toxic shock syndrome toxin-1 (TSST 1) was tested by high-performance methods. This preparation was homogenous in ion exchange chromatography and isoelectric focusing (pI = 7.4), but was resolved into two distinct peaks by high-performance hydroxyapatite chromatography. Both components, TSST-1hA and TSST-1hB had similar molecular weights (22 kD) and amino acid compositions. TSST-1 did not dimerize or polymerize upon heating at 60 degrees C for 30 min or in solutions with pH varying from 4.0 to 8.5. TSST-1hA and TSST-1hB showed similar immunological reactivity to native TSST-1 goat polyclonal antibodies. TSST-1hA and TSST-1hB as well as staphylococcal enterotoxin A and staphylococcal exfoliative toxin were potent mitogens in lymphocyte proliferation assays. The lymphocyte proliferative response to 10 pg of TSST-1hB was comparable to a response elicited by 10 ng of TSST-1hA, suggesting that the former component is a more potent mitogen. Rabbit or goat polyclonal antibodies to native TSST-1 efficiently neutralized both TSST-1 components. Heat treatment at 80 degrees C for 15 min had minimal or no effect on the mitogenic properties of TSST-1hA and TSST-1hB. PMID- 2558693 TI - Trends in incidence and results of treatment of testicular germ cell tumours in Finland 1972-1983. AB - A nationwide series of 422 patients with testicular germ cell cancer diagnosed in Finland in 1972-1983 was analysed. The age-adjusted incidence rate, although very low (1.6 per 10(5) male population per year), has increased compared to that in previous decades. The 3-year survival rate has improved markedly and was during the last part of the period high in patients with local (stage I) and regional (stages IIA-B) disease (100% for seminoma and over 90% for non-seminoma patients) but still fairly poor in advanced stages (stages IIC-IV) (58% for seminoma and 26% for non-seminoma patients). The improvement of the survival rate was most marked in non-seminoma patients below the median age (28.5 years). Cisplatin based chemotherapy was one of the major reasons for the improved prognosis, not only in non-seminoma patients but also in those with seminoma. There was no trend with time concerning the stage distribution of the disease. On the basis of relapse rates in stage I non-seminoma and seminoma patients staged surgically and clinically respectively, accuracy of clinical staging but not of surgical staging seemed to have improved. During the early period surgically staged stage I-II non seminoma patients had a slightly better prognosis than clinical stage I-II patients but a similar difference was not observed during the cisplatin era. PMID- 2558694 TI - Chromosome analysis of in situ breast cancer. AB - Eleven ductal carcinoma in situ (DCIS), 5 DCIS with microinvasion and 4 benign lesions have been investigated cytogenetically. Twelve of the 20 breast tumors (60%) had sufficient mitotic activity for chromosome analysis. All ductal carcinoma in situ had abnormal karyotypes, and clonal marker chromosomes could be identified in all tumors analyzed. None were cytogenetically normal. All but one of the 12 DCIS showed genetic heterogeneity. Tumor progression seems to be associated with loss of chromosomes. PMID- 2558695 TI - The treatment of in situ breast cancer. AB - Carcinoma in situ is the earliest histologically recognisable form of malignancy and as such provides an opportunity to treat the disease in a curative way. However, due to the comparative rarity of in situ breast carcinoma, there is no available information derived from controlled clinical trials. The two major variants, ductal carcinoma in situ (DCIS) and lobular carcinoma in situ (LCIS) will be considered separately as the two conditions have divergent natural histories. DCIS is increasing in incidence since microcalcification, which is a frequent accompaniment, may be detected radiologically in the screening of asymptomatic women. The extent of microcalcification may not indicate the extent of disease. It has yet to be determined whether there is a difference in behaviour of the tumour forming and the asymptomatic types of DCIS. After a biopsy has shown DCIS there will be residual DCIS at the biopsy site in one-third of patients, and multifocal DCIS in another third. A coexistent infiltrating carcinoma may be present in up to 16%. Due to sampling problems areas of invasion may be missed. Axillary nodal metastases are found in only 1% of patients with histological DCIS. Radical surgery by total or modified mastectomy is almost curative, but 3% of patients will die of metastases. Taking results of uncontrolled trials, local relapse rates are as follows: excision alone 50%, wide excision 30%, wide excision plus radiotherapy 20%. Two prospective trials are underway run by the EORTC and NSABP in which patients with DCIS are treated by wide excision with or without external radiotherapy. LCIS is usually an incidental finding with a bilateral predisposition to subsequent infiltrating carcinomas. Curative procedures such as bilateral mastectomy with reconstruction may represent overtreatment. A systemic rather than local approach would seem appropriate and a trial is now underway run by the EORTC in which patients with histologically confirmed LCIS are randomised to observation alone or to receive tamoxifen 20 mg daily for 5 years. Cooperative studies will provide the way of acquiring important data on treatment regimens of both DCIS and LCIS. It is timely that treatment regimens for in situ carcinoma of the breast be examined by controlled clinical trials. PMID- 2558696 TI - National surveillance of dialysis-associated diseases in the United States, 1987. AB - The Centers for Disease Control surveyed 1,630 chronic hemodialysis centers in the United States in 1987 in conjunction with the annual facility survey done by the Health Care Financing Administration. Information was obtained on the following diseases and practices: 1) hepatitis B virus (HBV) infection in patients and staff members; 2) infection control procedures for hepatitis B surface antigen (HBsAg)-positive patients; 3) frequency of HBsAg serologic screening; 4) use of hepatitis B vaccine; 5) non-A, non-B hepatitis in patients and staff members; 6) pyrogenic reactions and septicemia; 7) dialysis dementia; 8) new dialyzer syndrome; 9) high flux dialysis; 10) reuse of dialyzers, dialyzer caps, bloodlines, transducer filters; 11) cleaning and disinfection procedures; and 12) human immunodeficiency virus (HIV) infection. The response rate to a mailed questionnaire was 91%. These 1,486 centers represented 97,225 patients and 27,123 staff members. During the last 12 years, the incidence of HBV infection decreased from 3.0% to 0.2% among patients and from 2.6% to 0.1% among staff members. Over the same time, the prevalence of HBsAg-positivity declined from 7.8% to 1.7% among patients and from 0.9% to 0.4% among staff members. Hepatitis B vaccine was given by 88% of the centers. By the end of 1987, 14% of susceptible patients and 49% of susceptible staff members had received all three doses of hepatitis B vaccine. From 1982 to 1987, as a result of receiving vaccine, the prevalence of antibody to HBsAg (anti-HBs) increased from 12% to 18% among patients and from 18% to 50% among staff. The incidence of non-A, non-B hepatitis in 1987 was reported to be 1.2% among patients and 0.2% among staff members. Fourteen percent of the centers reported pyrogenic reactions among their patients, and 46% reported septicemia in the absence of pyrogenic reactions. Pyrogenic reactions were significantly more likely to be reported by centers that practiced high flux dialysis. The reported incidence of dialysis dementia among hemodialysis patients was 0.2%, with a case fatality rate of 29%. Among patients developing dialysis dementia, the case fatality rate was higher in those centers that used deionization (DI) without reverse osmosis (RO) (47%) compared with centers that used RO (28%) (not significant, p greater than 0.05). In 1987, 64% of centers reported that they reused disposable dialyzers. These centers treated 70% of the dialysis patient population.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2558697 TI - Clinical trials with a new iodinated lipid emulsion for computed tomography of the liver. AB - A new iodinated lipid emulsion, Intraiodol, for which animal studies have indicated better tolerance than for other iodinated lipid emulsions, was tested in 15 patients with malignant lesions, and in one patient with focal nodular hyperplasia. Repeated CT scans of the liver and spleen and blood tests were performed for 24 hours after intravenous injection of Intraiodol. The uptake of Intraiodol in the liver (peak mean 28.6 HU) was higher than in the spleen (peak mean 21.8 HU). The uptake of Intraiodol in malignant lesions was minimal (peak mean 2.8 HU). The detection rate of hepatic lesions was equal to or better than that achieved by US, CT, and/or CT angiography. However, liver uptake of Intraiodol was low in 2 patients with severe fatty infiltration. Intraiodol produced vascular enhancement up to one hour after injection since it was eliminated slowly from the circulation. The observed adverse reactions consisted of temporary metallic taste in 5 of the patients, fever and exacerbation of back pain in one patient, and transient thrombocytopenia in one patient. Alkaline phosphatase increased (17%, p less than 0.01) only at two hours, and erythrocyte count (6%, p less than 0.05) at 24 hours after injection. Our initial results indicate diagnostic advantages of Intraiodol without serious adverse reactions. Further clinical studies are required to confirm these findings. PMID- 2558698 TI - Computed tomography of adrenal metastases in hepatocellular carcinoma. Report of four cases. AB - Four patients with adrenal metastases from hepatocellular carcinoma (HCC) diagnosed with computed tomography are described. In three, the metastases showed a central area of low density due to tumor necrosis, irregular contrast enhancement, and a well-defined margin. They measured 8 cm or more in diameter. In one, the adrenal metastasis was at first a relatively small homogeneously solid mass, but later, when it grew larger, a central low density developed. Since the adrenal gland is the second most common site of metastasis from HCC at autopsy, a combination of an adrenal tumor and a liver tumor should suggest this possibility. PMID- 2558699 TI - Comparative potency of pipecuronium bromide and pancuronium bromide. AB - Cumulative dose-response curves were constructed to determine the comparative potency of pipecuronium and pancuronium. From these, the ED50 and ED95 values were calculated. These were 24.96 micrograms kg-1 and 44.96 micrograms kg-1, respectively, for pipecuronium and 30.42 micrograms kg-1 and 61.12 micrograms kg 1, respectively, for pancuronium. PMID- 2558702 TI - Prevention of deep vein thrombosis--use of the low molecular weight heparin enoxaparin. PMID- 2558701 TI - Beta-adrenoceptor responses to inhaled salbutamol in the elderly. AB - The purpose of the present study was to evaluate and compare the responsiveness of beta 2-adrenoceptors in elderly and young subjects. Seven healthy elderly volunteers (72 +/- 3 years) were given cumulative doses of inhaled salbutamol (100 micrograms-4000 micrograms) or placebo, following pre-treatment with propranolol 40 mg or placebo. Finger tremor (Tr), plasma potassium (K), and heart rate (HR) were measured at each dose step. There were dose-dependent increases in Tr (P less than 0.001) and HR (P less than 0.001) and falls in K (P less than 0.001), which were completely attenuated by propranolol (P less than 0.001). Comparison with dose-response curves in a group of young (Y) subjects (24 +/- 3 years) given an identical dose protocol of salbutamol showed no evidence of subsensitivity of beta 2-adrenoceptor responses in the elderly (E) group (mean and 95% confidence intervals for maximum responses): delta K -0.90 (-1.1(-)-0.82) mmol l-1 Y, -0.82 (-1.04(-)-0.60) mmol l-1 E, delta Tr 274 (213-335)% Y, 269 (197 342)% E, delta HR 25 (21-28) beats min-1 Y, 26 (21-31) beats min-1 E. PMID- 2558700 TI - Altered adrenoceptor responsiveness during adrenaline infusion but not during mental stress: differences between receptor subtypes and tissues. AB - 1. Effects of 3 h infusions of adrenaline (0.4 nmol kg-1 min-1) or placebo and of mental stress evoked by a colour word test (CWT) on adrenergic receptor function were investigated in healthy men. Responses of heart rate, blood pressure, plasma catecholamines, plasma cyclic AMP and plasma free fatty acids (FFA) were evaluated during infusions and CWT. In vitro beta 2-adrenoceptor numbers [( 125I] HYP binding) and function (isoprenaline induced cyclic AMP accumulation) were studied on lymphocytes in all experiments. alpha 2-adrenoceptor binding [( 3H] yohimbine and adrenaline) to intact platelets was evaluated in the infusion experiments only. 2. Placebo infusion evoked no major alterations of any parameter. 3. Adrenaline infusion raised venous plasma adrenaline levels to 4-5 nmol l-1, increased heart rate by 14 +/- 3 beats min-1 and plasma cyclic AMP by 17 +/- 3 nmol l-1, and decreased diastolic blood pressure by 15 +/- 5 mm Hg. These responses persisted throughout the infusion. Plasma FFA levels, on the other hand, increased at 30 min of infusion (from 236 +/- 44 to 717 +/- 92 mumol l-1) and returned to basal levels after 3 h of infusion. 4. In vitro, lymphocytes showed increased beta 2-responsiveness after 30 min of adrenaline infusion (delta cyclic AMP increased from 1.86 +/- 0.24 to 3.06 +/- 0.58 pmol/10(6) cells), but a decreased response (0.47 +/- 0.10 pmol/10(6) cells) after 3 h of infusion. [125I] HYP binding to lymphocyte membranes showed a three-fold increase of Bmax at 30 min of adrenaline infusion followed by a return to basal values after 3 h of infusion. [125I]-HYP binding reflected the functional responsiveness of the lymphocytes in vitro poorly. alpha 2-adrenoceptors on platelets were not altered with regard to Bmax or Kd for [3H]-yohimbine binding or Ki for adrenaline displacement of [3H]-yohimbine binding. 5. CWT evoked marked circulatory changes, a four-fold increase in plasma adrenaline and a 60% increase in beta 2 adrenoceptor binding sites without changes in functional responsiveness of the lymphocytes. 6. We conclude that exposure to high physiological levels of adrenaline in vivo alters lymphocyte beta-adrenoceptor responsiveness in a biphasic manner, with an early increase followed by a later decrease, but that most beta-adrenoceptor mediated responses to adrenaline in vivo remain intact. Lymphocyte alterations may reflect recruitment of cells into the circulation during sympathoadrenal stimulation. Platelet alpha 2-adrenoceptors are apparently not easily subjected to agonist induced dynamic receptor regulation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2558703 TI - Polyclonal rearrangements of the T-cell receptor beta-chain in fatal angioimmunoblastic lymphadenopathy. AB - Genomic rearrangement of germline T-cell antigen receptor (TcR) and immunoglobulin (Ig) genes was studied by Southern blot analysis in seven patients with angioimmunoblastic lymphadenopathy (AILD). In three cases clinically suspected of transformation into malignant lymphoma, hybridization with the TcR beta probe showed markedly dimished intensity in the 11.5 kb germline band after Eco RI digestion and normal germline configuration after Hind III and Bam HI digestion, indicating polyclonal T cell rearrangements. A clonal rearrangement of the TcR beta gene was detected in only one case at initial biopsy. No monoclonal rearrangement of Ig genes was observed. These data show that in some cases of AILD disease progression is indicated by polyclonal TcR rearrangements and not by outgrowth of a malignant clone, supporting the concept of AILD as an immunoregulatory disorder. PMID- 2558704 TI - Pure red cell aplasia in association with virus associated haemophagocytic syndrome (VAHS). PMID- 2558706 TI - The choice of suture materials and techniques for repair of perineal trauma: an overview of the evidence from controlled trials. AB - A systematic review of the 14 relevant controlled trials was conducted because there is no agreement about the choice of material and technique for repair of perineal trauma sustained during childbirth. Derivatives of polyglycolic acid (marketed as Dexon and Vicryl) appear to be the absorbable material of choice for both deep and skin closure. Compared with catgut their use is associated with about a 40% reduction in short-term pain and need for analgesia. The main drawback is that some material often needs removal during the puerperium. Glycerol-impregnated catgut is ruled out because of its link with long-term dyspareunia. Compared with the non-absorbable materials (silk and nylon) polyglycolic acid skin sutures were associated with less short-term perineal pain, and had no clear disadvantages. Continuous, subcuticular stitching appears preferable to interrupted, transcutaneous suturing, particularly in terms of perineal pain in the early puerperium. PMID- 2558705 TI - The Southmead perineal suture study. A randomized comparison of suture materials and suturing techniques for repair of perineal trauma. AB - Commonly used suture materials and techniques for perineal repair following vaginal delivery were compared in a randomized controlled trial involving 1574 women. Three comparisons were made using a modified factorial design. In the comparison of teflon-coated polyglycolic acid (Dexon plus) with chromic catgut for repair of the vagina and deep perineal tissues there was no clear difference other than less short-term analgesia being required in association with polyglycolic acid. Outcome was also similar after skin repair with either polyglycolic acid or chromic catgut or silk, although silk repair required more packets of material and was associated with delay in resuming sexual intercourse; polyglycolic acid was more likely to need removal than chromic catgut but it appeared to reduce the need for resuturing. There was no clear difference between continuous subcuticular and interrupted transcutaneous sutures for repair of perineal skin. PMID- 2558707 TI - Addisonian crisis in pregnancy. Case report. PMID- 2558709 TI - Long-range electron transfer in multisite metalloproteins. PMID- 2558708 TI - Long-term outpatient treatment of CMV retinitis with ganciclovir in AIDS patients. AB - Cytomegaloviral retinitis was diagnosed in nine eyes of seven patients with acquired immune deficiency syndrome (AIDS) on the basis of the characteristic ocular findings and a positive culture for cytomegalovirus (CMV) obtained systemically. Treatment with ganciclovir was begun on a protocol which provided two weeks of inpatient therapy at daily doses of 2.5 to 10 mg/kg followed by outpatient therapy at a reduced dosage three to six days per week. Outpatient maintenance dosage ranged from 15 mg/kg per week to 30 mg/kg per week. In seven eyes of six patients the treatment decreased retinal inflammation and stabilised the margins of the lesions. Six patients have tolerated long term maintenance therapy for 10-30 weeks. Six of seven patients (85%) in this study developed side effects from ganciclovir which required periods of a reduction in dosage or interruption of therapy. The side effects from ganciclovir included neutropenia, thrombocytopenia, drug fever, and neuropathy. Physicians using ganciclovir in AIDS patients must watch for the signs of drug toxicity and adjust treatment accordingly. Ganciclovir appears to be a promising therapy for CMV retinitis, but further work is necessary to determine the best regimen for optimal efficacy with minimal side effects. PMID- 2558710 TI - Identification by proton nuclear magnetic resonance of the histidines in cytochrome b5 modified by diethyl pyrocarbonate. AB - Diethyl pyrocarbonate (DEP) is an electrophilic reagent that is used to modify reversibly the histidine residues of proteins. Unfortunately, the lability of the acylated histidine adduct usually does not permit the isolation and identification of the modified histidine. By use of 500-MHz proton NMR spectroscopy, it has been possible to identify the C-H resonances of the nonaxial histidines of trypsin-solubilized bovine, rabbit, and porcine cytochrome b5 and therefore observe the interaction of DEP with specific histidine residues of cytochrome b5. In addition, the pKa of the peripheral histidines of bovine and rabbit cytochrome b5 have been measured in D2O. In the bovine protein it was found that the histidines are modified sequentially with increasing DEP concentration in the order His-26 greater than His-15 greater than His-80. This order is maintained in the rabbit protein with the following additions: His-26 approximately His-27 greater than His-15 greater than or equal to His-17 greater than His-80. The relative reactivity of the peripheral histidines with DEP was rationalized by considering three of their characteristics: (1) the pKa of the histidine, (2) the fraction of the side chain exposed to the solvent, and (3) the hydrogen-bond interactions of the imidazole ring. PMID- 2558712 TI - Structural studies on transmembrane proteins. 2. Spin labeling of bacteriorhodopsin mutants at unique cysteines. AB - Site-directed mutagenesis was used to produce mutants of bacteriorhodopsin where either glycine-72, threonine-90, leucine-92, or serine-169 was replaced by a cysteine. Two different spin labels were then covalently attached to these sites. The selection of attachment sites covered two postulated loops (72,169) and a membrane-spanning segment (90,92). It was not possible to properly refold the protein labeled at position 90, presumably due to steric problems, but the EPR spectra of the other mutants that were successfully reconstituted in phospholipid vesicles provided information on the dynamics of protein side chains in the vicinity of the label site. A power saturation approach was used to investigate the spin relaxation times, which in turn can be influenced by collisions with paramagnetic species. The differential effect of oxygen and a water-soluble chromium complex on the power-saturation behavior of the spin-labeled mutants was used to obtain topographical information on the sites in the membrane-bound protein. The results are consistent with residues 72 and 169 being located in structured loops exposed to the aqueous phase and residue 92 being localized in the membrane interior, possibly near a helix-helix contact region. PMID- 2558711 TI - Castanospermine inhibits the function of the low-density lipoprotein receptor. AB - Castanospermine, a plant alkaloid that inhibits the glycoprotein processing enzyme glucosidase I, has been used to inhibit N-linked oligosaccharide modification, resulting in the production of glycoproteins having Glc3Man7 9(GlcNAc)2 oligosaccharides. This alkaloid caused a significant inhibition of LDL endocytosis in cultured primate smooth muscle cells and human skin fibroblasts. At an optimum concentration of 250 micrograms/mL, castanospermine caused a 40% decrease in cell surface receptor-mediated LDL binding at 4 degrees C, with no apparent change in affinity. Further, the inhibitor had no direct effect on LDL metabolism. This inhibition of LDL receptor expression and function occurred only when the drug was present during de novo receptor synthesis, i.e., during up regulation. Although the number of cell surface LDL receptors was significantly reduced in the presence of castanospermine, the total number of receptors in the cell was only slightly reduced, indicating that castanospermine induced a redistribution rather than a reduction in the number of receptors. Similarly, subcellular fractionation studies confirmed that castanospermine treatment of fibroblasts results in an altered distribution of receptor activity compared with controls. These findings are consistent with the conclusion that the decrease in specific LDL binding to cells grown in the presence of castanospermine is due to intracellular redistribution of the LDL receptor so that more receptor remains in internal compartments as a result of a diminished rate of transport. PMID- 2558713 TI - Subunit composition of the purified dihydropyridine binding protein from skeletal muscle. AB - The dihydropyridine (DHP) receptor from rabbit skeletal muscle has been characterized by affinity labeling and purification. Two procedures were used for purification: one that was a procedure modified from that of Curtis and Catterall (1984) and one that employed an anti alpha 1 monoclonal antibody (Mab) affinity column. In addition, both digitonin and CHAPS solubilizations were utilized with each purification technique. The major findings are as follows: (1) In contrast to the behavior in digitonin, neither the 52K (beta) nor the 140K (alpha 2) polypeptide quantitatively copurifies with the 170K (alpha 1) polypeptide when the purification is carried out in CHAPS. This has been shown by use of both wheat germ and monoclonal antibody columns. The digitonin-extracted receptor complex bound to the Mab affinity column loses alpha 2 and beta when the digitonin is replaced by CHAPS, and when the complex is bound to a WGA column, a CHAPS wash causes dissociation of alpha 1, beta, and gamma from alpha 2. Loss of binding of dihydropyridines occurs with the CHAPS wash but can be partially restored by the addition of the CHAPS wash to the material eluted from the column with N-acetylglucosamine. (2) Although both detergents solubilized greater than 80% of the polypeptides associated with the DHP binding site, the ability of these proteins to bind dihydropyridines is reduced more by CHAPS treatment than by digitonin treatment, raising the possibility that subunit interactions contribute to high-affinity binding. Alternatively, CHAPS may remove tightly bound lipids necessary for binding or cause irreversible denaturation of the binding site.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558714 TI - Contribution of a single hydroxyl group to transition-state discrimination by adenosine deaminase: evidence for an "entropy trap" mechanism. AB - Adenosine deaminase was found to bind 6-hydroxy-1,6-dihydropurine ribonucleoside (II), formed by reversible addition of water to purine ribonucleoside (I) in a reaction analogous to formation of a tetrahedral intermediate in substrate deamination, with an apparent Ki value of 3 x 10(-13) M at 20 degrees C. 1,6 Dihydropurine ribonucleoside (IV), synthesized by photolysis of purine ribonucleoside in the presence of NaBH4, exhibited a Ki value of 5.4 x 10-6 M. After correction for differences between the relative free energies of solvation of II and IV, the 6-hydroxyl group of II was estimated to contribute more than 16 kcal to the free energy of binding, approaching the enthalapy of formation of a single hydrogen bond to charged group in the vapor phase. The relatively weak binding of IV and of substrate water suggests that entropic effects, arising from the cooperative action of binding determinants contained within these separate molecules, contribute more than 10 kcal/mol to the free energy of binding of II in which these binding determinants are contained within a single molecule. In free solution, the entropy of reversible hydration of I was evaluated by measuring the temperature dependence of equilibria of protonation of I and of pseudobase formation from I-methylpurinium ribonucleoside as -35 eu, comparable with the entropy of activation for the uncatalyzed hydrolysis of adenosine. In the active site of adenosine deaminase, this thermodynamic obstacle is evidently climbed spontaneously as a result of attractive interactions between the active site and the critical hydroxyl group at the 6-position.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558715 TI - Identification of the site on calcineurin phosphorylated by Ca2+/CaM-dependent kinase II: modification of the CaM-binding domain. AB - The catalytic subunit of the Ca2+/calmodulin- (CaM) dependent phosphoprotein phosphatase calcineurin (CN) was phosphorylated by an activated form of Ca2+/CaM dependent protein kinase II (CaM-kinase II) incorporating approximately 1 mol of phosphoryl group/mol of catalytic subunit, in agreement with a value previously reported (Hashimoto et al., 1988). Cyanogen bromide cleavage of radiolabeled CN followed by peptide fractionation using reverse-phase high-performance liquid chromatography yielded a single labeled peptide that contained a phosphoserine residue. Microsequencing of the peptide allowed both the determination of the cleavage cycle that released [32P]phosphoserine and the identity of amino acids adjacent to it. Comparison of this sequence with the sequences of methionyl peptides deduced from the cDNA structure of CN (Kincaid et al., 1988) allowed the phosphorylated serine to be uniquely identified. Interestingly, the phosphoserine exists in the sequence Met-Ala-Arg-Val-Phe-Ser(P)-Val-Leu-Arg-Glu, part of which lies within the putative CaM-binding site. The phosphorylated serine residue was resistant to autocatalytic dephosphorylation, yet the slow rate of hydrolysis could be powerfully stimulated by effectors of CN phosphatase activity. The mechanism of dephosphorylation may be intramolecular since the initial rate was the same at phosphoCN concentrations of 2.5-250 nM. PMID- 2558716 TI - Vanadate and molybdate increase tyrosine phosphorylation in a 50-kilodalton protein and stimulate secretion in electropermeabilized platelets. AB - Addition of vanadate and molybdate to electropermeabilized human platelets caused a time- and dose-dependent increase in the phosphotyrosyl content of 50- and 38 kDa proteins. This effect can most likely be attributed to an inhibition of protein-tyrosine-phosphatase activity because vanadate and molybdate inhibited this activity in platelet extracts by greater than 97% while causing an increase in tyrosyl phosphorylation of artificial substrates that had been added to the same extracts. The addition of vanadate and molybdate to the electropermeabilized platelets also induced an increase in serotonin and PDGF secretion. Interestingly, the secretion of these components tightly correlated in a time- and dose-dependent fashion with the phosphorylation of the 50-kDa protein on tyrosyl residues. This suggests that the tyrosine phosphorylation of this protein may be closely linked to the platelet activation cascade. PMID- 2558717 TI - 31P and 1H NMR studies of the structure of enzyme-bound substrate complexes of lobster muscle arginine kinase: relaxation measurements with Mn(II) and Co(II). AB - The paramagnetic effects of Mn(II) and Co(II) on the spin-lattice relaxation rates of 31P nuclei of ATP and ADP and of Mn(II) on the spin-lattice relaxation rate of the delta protons of arginine bound to arginine kinase from lobster tail muscle have been measured. Temperature variation of 31P relaxation rates in E.MnADP and E.MnATP yields activation energies (delta E) in the range 6-10 kcal/mol. Thus, the 31P relaxation rates in these complexes are exchange limited and cannot provide structural information. However, the relaxation rates in E.CoADP and E.CoATP exhibit frequency dependence and delta E values in the range 1-2 kcal/mol; i.e., these rates depend upon 31P-Co(II) distances. These distances were calculated to be in the range 3.2-4.5 A, appropriate for direct coordination between Co(II) and the phosphoryl groups. The paramagnetic effect of Mn(II) on the 1H spin-lattice relaxation rate of the delta protons of arginine in the E.MnADP.Arg complex was also measured at three frequencies (viz., 200, 300, and 470 MHz). These 1H experiments were performed in the presence of sufficient excess of arginine to be observable over the protein background but with MnADP exclusively in the enzyme-bound form so that the enhancement in the relaxation rates of the delta protons of arginine arises entirely from the enzyme-bound complex. Both the observed frequency dependence of these rates and the delta E less than or equal to 1.0 +/- 0.3 kcal/mol indicate that this rate depends on the 1H-Mn(II) distances.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558718 TI - 1H NMR structural characterization of a recombinant kringle 2 domain from human tissue-type plasminogen activator. AB - The kringle 2 domain of human tissue-type plasminogen activator (t-PA) has been characterized via 1H NMR spectroscopy at 300 and 620 MHz. The experiments were performed on the isolated domain obtained by expression of the 174-263 portion of t-PA in Escherichia coli [Cleary et al. (1989) Biochemistry 28, 1884-1891]. The spectrum of t-PA kringle 2 is characteristic of a globular structure and shows overall similarity to that of the plasminogen (PGN) kringle 4. Spectral comparison with human and bovine PGN kringle 4 identifies side-chain resonances from Leu46, which afford a fingerprint of kringle folding, and from most of the aromatic ring spin systems. Assignment of signals arising from the His13, His48a, and His64 side chains, which are unique to t-PA kringle 2, was assisted by the availability of a His64----Tyr mutant. Ligand-binding studies confirm that t-PA kringle 2 binds L-lysine with an association constant Ka approximately 11.9 mM-1. The data indicate that homologous or conserved residues relative to those that compose the lysine-binding sites of PGN kringles 1 and 4 are involved in the binding of L-lysine to t-PA kringle 2. These include Tyr36 and, within the kringle inner loop, Trp62, His64, Trp72, and Tyr74. Acid/base titration of aromatic singlets in the presence of L-lysine yields pKa* approximately 6.25 and approximately 4.41 for His13 and His64, respectively, and shows that the His48a imidazole group does not protonate down to pH* approximately 4.3. Thus, the His48a and His64 side chains are in solvent-shielded locations. As observed for the PGN kringles, the Trp62 indole group titrates with pKa* approximately 4.60, which indicates proximity of the side chain to a titratable carboxyl group, most likely that of Asp57 at the binding site. Several labile NH protons of t-PA kringle 2 exhibit retarded H-exchange kinetics, requiring more than a week in 2H2O for full deuteration in the presence of L-lysine at 37 degrees C. This reveals that kringle 2 is endowed with a compact, dynamically stable conformation. Proton Overhauser experiments in 1H2O, centered on well-resolved NH resonances between 9.8 and 12 ppm, identify signals arising from the His48a imidazole NH3 proton and the three Trp indole NH1 protons. A strong dipolar interaction was observed among the Trp25 indole NH1, the Tyr50 amide NH, and the His48a imidazole CH2 protons, which affords evidence for an aromatic cluster in t PA kringle 2 similar to that found at the hydrophobic kernel of PGN kringles.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2558719 TI - Interplay of phosphorylation and dephosphorylation in vision: protein phosphatases of bovine rod outer segments. AB - Two types of protein phosphatases were identified in carefully prepared bovine rod outer segments (ROS). Extraction of the ROS with a medium-salt buffer solubilized protein phosphatase activity that was mainly type 2A, since it was active toward phosphorylase a in the absence of divalent cations, was not retained by heparin-Sepharose, dephosphorylated the alpha-subunit of phosphorylase kinase faster that the beta-subunit, and was unaffected by inhibitor 2. Further extraction of the resulting membranes with a high-salt buffer solubilized additional phosphatase activity which was predominantly type 1, since it was retained by heparin-Sepharose and was blocked by inhibitor 2. The molecular mass of the type 2A phosphatase estimated by gel permeation chromatography on Superose 12 was 100 kDa, suggesting it may be the 2A2 form. Only the ROS type 2A phosphatase dephosphorylated opsin and rhodopsin efficiently. Concordant with this finding, the purified catalytic subunit of protein phosphatase 2A from rabbit skeletal muscle dephosphorylated opsin efficiently, while the type 1 catalytic subunit isolated from this tissue was inactive. Together, the results suggest that the ROS type 2A protein phosphatase plays an important role in regenerating rhodopsin from the various phosphorylated species in vivo. The activity of the enzyme per retina (approximately 85 pmol of Pi released/min) is comparable to that of rhodopsin kinase (100 pmol of phosphate transferred/min). PMID- 2558720 TI - A calcium-specific site influences the structure and activity of the manganese cluster responsible for photosynthetic water oxidation. AB - EPR studies have revealed that removal of calcium using citric acid from the site in spinach photosystem II which is coupled to the photosynthetic O2-evolving process produces a structural change in the manganese cluster responsible for water oxidation. If done in the dark, this yields a modified S1' oxidation state which can be photooxidized above 250 K to form a structurally altered S2' state, as seen by formation of a "modified" multiline EPR signal. Compared to the "normal" S2 state, this new S2'-state EPR signal has more lines (at least 25) and 25% narrower 55Mn hyperfine splittings, indicative of disruption of the ligands to manganese. The calcium-depleted S2' oxidation state is greatly stabilized compared to the native S2 oxidation state, as seen by a large increase in the lifetime of the S2' EPR signal. Calcium reconstitution results in the reduction of the oxidized tyrosine residue 161YD+ (Em approximately 0.7-0.8 V, NHE) within the reaction center D1 protein in both the S1' and S2' states, as monitored by its EPR signal intensity. We attribute this to reduction by Mn. Thus a possible structural role which calcium plays is to bring YD+ into redox equilibrium with the Mn cluster. Photooxidation of S2' above 250 K produces a higher S state (S3 or S4) having a new EPR signal at g = 2.004 +/- 0.003 and a symmetric line width of 163 +/- 3 G, suggestive of oxidation of an organic donor, possibly an amino acid, in magnetic contact with the Mn cluster. This EPR signal forms in a stoichiometry of 1-2 relative to YD+.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558721 TI - Sequence- and structure-dependent DNA base dynamics: synthesis, structure, and dynamics of site and sequence specifically spin-labeled DNA. AB - A nitroxide spin-labeled analogue of thymidine (1a), in which the methyl group is replaced by an acetylene-tethered nitroxide, was evaluated as a probe for structural and dynamics studies of sequence specifically spin-labeled DNA. Residue 1a was incorporated into synthetic deoxyoligonucleotides by using automated phosphite triester methods. 1H NMR, CD, and thermal denaturation studies indicate that 1a (T*) does not significantly alter the structure of 5' d(CGCGAATT*CGCG) from that of the native dodecamer. EPR studies on monomer, single-stranded, and duplexed DNA show that 1a readily distinguishes environments of different rigidity. Comparison of the general line-shape features of the observed EPR spectra of several small duplexes (12-mer, 24-mer) with simulated EPR spectra assuming isotropic motion suggests that probe 1a monitors global tumbling of small duplexes. Increasing the length of the DNA oligomers results in significant deviation from isotropic motion, with line-shape features similar to those of calculated spectra of objects with isotropic rotational correlation times of 20-100 ns. EPR spectra of a spin-labeled GT mismatch and a T bulge in long DNAs are distinct from those of spin-labeled Watson-Crick paired DNAs, further demonstrating the value of EPR as a tool in the evaluation of local dynamic and structural features in macromolecules. PMID- 2558722 TI - Highly efficient DNA delivery mediated by pH-sensitive immunoliposomes. AB - We have previously shown that pH-sensitive immunoliposomes can mediate a target specific delivery of plasmid DNA to tumor cells grown in a mouse model [Wang, C. Y., & Huang, L. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 7851-7855]. The efficiency of delivery in terms of the target cell transformation frequency has now been characterized for both short- and long-term gene expression in a tissue culture system. Herpes simplex virus thymidine kinase (TK) gene was used as a reporter gene. It was placed under the control of the promoter for the rat phosphoenolpyruvate carboxykinase gene, which contains a cAMP regulatory element. Therefore, the expression of the exogenous gene in the target cell, mouse Ltk- cells, can be regulated by cAMP drugs. The plasmid DNA was encapsulated in liposomes using a detergent dialysis method. The efficiency of gene delivery was optimized with respect to the time course and dose of liposome-associated DNA. The existence of antibody of the liposomes was essential for the maximal level of DNA delivery. Delivery was also dependent on the lipid composition of the liposome. The pH-sensitive lipid composition gave 8-fold higher efficiency than the corresponding pH-insensitive composition. The transformation efficiency of the target cell also depended on the regulation of gene expression; cells incubated with dibutyryl-cAMP and theophylline showed a much higher level of transformation frequency than cells incubated without the drugs. When all liposome and incubation parameters are optimized, the Ltk- cells showed a 47% efficiency for the short-term transformation, and 2% for the long-term transformation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558723 TI - Substrate properties of 25-nt parallel-stranded linear DNA duplexes. AB - Four 25-nt oligonucleotides consisting of sequences of dA and dT (D1-4) have been synthesized. As shown in a companion paper (Rippe et al., 1989), the two combinations D1.D3 and D2.D4 form normal antiparallel duplexes, whereas the pairs D1.D2 and D3.D4 constitute duplexes with the same sequences, but with the two strands parallel to each other. The activities of the following DNA processing enzymes and chemical reagents on the parallel stranded (ps) and antiparallel stranded (aps) duplexes were tested. (i) The restriction endonucleases DraI, SspI, and MseI do not cut the ps duplexes. (ii) DNase I and exonuclease III exhibit a much lower activity with the ps duplexes. (iii) The nuclease activities of S 1 nuclease, micrococcal nuclease (S 7), phage lambda 5'-exonuclease, and the 3'-5' nuclease activity of Escherichia coli DNA polymerase I and its large fragment are higher with the ps than with the aps substrates. (iv) Bal 31 nuclease and the chemical nuclease 1,10-phenanthroline-copper ion [(OP)2Cu+] degrade ps-DNA and aps-DNA at approximately the same rate but show preferred cutting sites only with the aps molecules. (v) The iron(II)-EDTA complex has equivalent nuclease activities with the ps and the aps molecules. (vi) The ps duplex is not a substrate for blunt-end ligation with phage T4 DNA ligase. PMID- 2558724 TI - Spin-labeling proton NMR study on aromatic amino acid residues in the guanine nucleotide binding site of human c-Ha-ras(1-171) protein. AB - A truncated human c-Ha-ras gene product, ras(1-171) protein, was prepared and chemically modified with maleimide spin-label (MSL). By trypsin digestion of the MSL-labeled ras(1-171) protein, MSL-labeled peptide fragments were isolated and sequenced. The cysteine residue in position 118 of the protein, but not the other cysteine residues, Cys-51 or Cys-80, was found to be specifically labeled by MSL. The ESR spectrum of the MSL-labeled ras(1-171) protein indicates that the MSL group attached to Cys-118 is strongly immobilized. Proton NMR spectra at 400-MHz were measured for this MSL-labeled ras(1-171) protein and also for a control sample of a labeled ras(1-171) protein whose MSL was reduced by sodium ascorbate. In the difference spectra for these two proteins, resonances of protons in the vicinity of the MSL group attached to Cys-118 of the ras(1-171) protein were observed. Thus, the MSL group was found to be in the vicinity of the protein bound GDP. A phenylalanine residue and two histidine residues, which were characterized by 2D HOHAHA and DQF-COSY spectra, were also found to be in the vicinity of MSL. NOE and pH titration analyses indicate that this phenylalanine residue is close to the bound GDP and one of the two histidine residues. By carboxypeptidase digestion, the two histidine residues near MSL were identified as His-27 and His-94.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558725 TI - Thermostability of cytochrome c-552 from the thermophilic hydrogen-oxidizing bacterium Hydrogenobacter thermophilus. AB - The denaturation of the c-type cytochrome of the thermophilic bacterium Hydrogenobacter thermophilus cytochrome c-552 by heat and guanidine hydrochloride was studied by measuring the change in circular dichroic spectra. The melting temperature (T1/2) of cytochrome c-552 in the presence of 1.5 M guanidine hydrochloride was 34 degrees C higher than that of the c-type cytochrome of Pseudomonas aeruginosa cytochrome c-551. Hydrogenobacter cytochrome c-552 is a much more stable protein than cytochrome c-551 of the mesophilic bacterium P. aeruginosa, even though their amino acid sequences are 56% identical and they have numerous other similarities. However, notwithstanding these similarities between the sequences of the cytochromes c-552 and c-551 that were compared, it is very likely that these differences in stability could be due to some heretofore undefined differences in their spatial structures. It has been suggested that alpha-helix structure and electrostatic interaction could be the source of the stable spatial structure of cytochrome c-552. PMID- 2558726 TI - The substrate proton of the pyruvate kinase reaction. AB - The pyruvate kinase reaction occurs in separate phosphate- and proton-transfer stages: (formula; see text) K+, Mg2+, and Mg.ADP are known to be required for the phosphoryl transfer step, and K+ and Mg2+ with allosteric stimulation by MgATP are important for proton transfer. This paper uses the isotope trapping method with 3H-labeled water to identify the proton donor and determine when in the sequence of the catalytic cycle it is generated. When the enzyme was allowed to exchange briefly with 3H2O (pulse phase) and then diluted into a mixture containing PEP, ADP, and the cofactor K+, Mg2+, or Co2+ in D2O (chase phase), an amount of [3H]pyruvate was formed in great excess of the amount expected from steady-state catalysis in the diluted 3H-labeled water. With K+, Mg2+, and ADP at pH 6-9.5 in the pulse phase, a limit of 1.25 enzyme equiv of 3H were trapped. The concentration of PEP required for half-maximum trapping was 14-fold greater than its steady-state Km. Therefore, the rate constant for dissociation of the donor proton is estimated to be 14 times the steady-state rate of [3H]pyruvate formation, approximately 109 s-1, or 1500 s-1. At pD 6.4, Mg2+ and ADP were required in the chase, indicating that the ADP in the pulse was not bound tightly enough to be used in the chase. At pD 9.4, ADP was not required in the chase, only Mg2+ or Co2+, making it possible to limit the chase to one turnover from hybrid labeled complexes such as E.K.Mg.CoADP or E.K.Co.MgADP and PEP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558727 TI - Alternative proton donors/acceptors in the catalytic mechanism of the glutathione reductase of Escherichia coli: the role of histidine-439 and tyrosine-99. AB - The cloned Escherichia coli gor gene encoding the flavoprotein glutathione reductase was placed under the control of the tac promoter in the plasmid pKK223 3, allowing expression of glutathione reductase at levels approximately 40,000 times those of untransformed cells. This greatly facilitated purification of the enzyme. By directed mutagenesis of the gor gene, His-439 was changed to glutamine (H439Q) and alanine (H439A). The tyrosine residue at position 99 was changed to phenylalanine (Y99F), and in another experiment, the H439Q and Y99F mutations were united to form the double mutant Y99FH439Q. His-439 is thought to act in the catalytic mechanism as a proton donor/acceptor in the glutathione-binding pocket. The H439Q and H439A mutants retain approximately 1% and approximately 0.3%, respectively, of the catalytic activity of the wild-type enzyme. This reinforces our previous finding [Berry et al. (1989) Biochemistry 28, 1264-1269] that direct protonation and deprotonation of the histidine residue are not essential for the reaction to occur. The retention of catalytic activity by the H439A mutant demonstrates further that a side chain capable of hydrogen bonding to a water molecule, which might then act as proton donor, also is not essential at this position. Tyr-99 is a further possible proton donor in the glutathione-binding pocket, but the Y99F mutant was essentially fully active, and the Y99FH439Q double mutant also retained approximately 1% of the wild-type specific activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558728 TI - Inhibition of restriction endonuclease cleavage via triple helix formation by homopyrimidine oligonucleotides. AB - A 17-mer homopyrimidine oligonucleotide was designed to bind to the major groove of SV40 DNA at a 17 base pair homopurine-homopyrimidine sequence via Hoogsteen base pairing. This sequence contains the recognition site for the class II-S restriction enzyme Ksp 632-I. The oligonucleotide was shown to inhibit enzymatic cleavage under conditions that allow for triple helix formation. Inhibition is sequence-specific and occurs in the micromolar concentration range. Triple helix formation by oligonucleotides opens new possibilities for sequence-specific regulation of gene expression. PMID- 2558729 TI - Evidence for the formation of strand-break precursors in hydroxy-attacked thymidine 5'-monophosphate by the spin trapping method. AB - A method combining spin trapping, ESR, and HPLC was employed to obtain evidence for the formation of sugar radicals in OH-attacked TMP with special emphasis on the detection of strand-break precursors of DNA. OH radicals were produced by irradiating an N2O-saturated aqueous solution with X-rays. When an N2O-saturated aqueous solution containing TMP and a spin trapping reagent, MNP, was irradiated with X-rays, it was estimated on the basis of theoretical calculations using rate constants that 94% of the TMP radicals were induced by OH radicals. Since several spin adducts between TMP radicals and MNP, as well as the byproducts of the spin trapping reagent itself, were produced, reverse-phase HPLC was used to separate them. The presence of six spin adducts was confirmed by ESR examination. Further examination of these spin adducts by UV absorbance spectrophotometry showed the presence of a chromophore at 260 nm in three adducts. Since a gradual increase in the release of unaltered base from these adducts was observed when they were allowed to stand for 0-22 h at room temperature, they could be regarded as the spin adducts of sugar radicals and MNP. ESR spectra from the spin adducts were consistent with hydrogen abstraction radicals at the C1', C4', and C5' positions of the sugar moiety. These radicals appeared to be precursors of AP sites and strand breaks. In addition to these spin adducts, ESR spectra that were consistent with the spin adducts of base radicals (the C5 and C6 radicals) and MNP were observed. PMID- 2558730 TI - Replacement of a conserved proline and the alkaline conformational change in iso 2-cytochrome c. AB - Although point mutations usually lead to minor localized changes in protein structure, replacement of conserved Pro-76 with Gly in iso-2-cytochrome c induces a major conformational change. The change in structure results from mutation induced depression of the pK for transition to an alkaline conformation with altered heme ligation. To assess the importance of position 76 in stabilizing the native versus the alkaline structure, the equilibrium and kinetic properties of the pH-induced conformational change have been compared for normal and mutant iso 2-cytochrome c. The pKapp for the conformational change is reduced from 8.45 (normal iso-2) to 6.71 in the mutant protein (Gly-76 iso-2), suggesting that conservation of Pro-76 may be required to stabilize the native conformation at physiological pH. The kinetics of the conformational change for both the normal and mutant proteins are well-described by a single kinetic phase throughout most of the pH-induced transition zone. Over this pH range, a minimal mechanism proposed for horse cytochrome c [Davis, L. A., Schejter, A., & Hess, G. P. (1974) J. Biol. Chem. 249, 2624-2632] is consistent with the data for normal and mutant yeast iso-2-cytochromes c: NH KH----N + H+ kcf in equilibrium kcb A NH and N are native forms of cytochrome c with a 695-nm absorbance band, A is an alkaline form that lacks the 695-nm band, KH is a proton dissociation constant, and kcf and kcb are microscopic rate constants for the conformational change. The Gly-76 mutation increases kcf by almost 70-fold, but kcb and KH are unchanged.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558731 TI - [A method of determining fragments of peptide bioregulators responsible for interaction with receptors]. AB - A method for investigating the relationship between chemical structure of peptide molecules and their biological activity is suggested. It is based on a few statistical algorithms which are described. The results of the method testing on the thyrotropin-releasing hormone analogs are presented. PMID- 2558732 TI - [The effect of N-(1-methyldodecyl)-N,N-dimethylaminoxide on the conformation of hydrocarbon chains in phospholipid bilayers isolated from Escherichia coli]. AB - Using the method of ESR spectroscopy of stearic acid spin probes labeled by the doxyl group on the 12th or 16th carbon, it has been found that bactericidal surfactant N-(1-methyldodecyl)-N,N-dimethylamine oxide increases the effective energy difference between trans- and gauche conformers Eg and decreases the probability of gauche conformers formation pg in lipid hydrocarbon chains in multilamellar liposomes prepared from Escherichia coli-isolated phospholipids, at low surfactant concentrations. Above the surfactant: phospholipid molar ratio of 1:14 to 1:17, the value of Eg decreases and that of pg increases. The results are interpreted using the cluster model of lipid bilayer. At low concentrations the surfactant molecules are inserted into the dynamical defects between the clusters, thereby increasing the packing density of chains in the bilayer. At high concentrations the surfactant molecules penetrate into the clusters perturbing the dense packing of chains in clusters. PMID- 2558733 TI - Thermodynamics of phosphatidylcholine-cholesterol mixed model membranes in the liquid crystalline state studied by the orientational order parameter. AB - It is shown that good estimates of the activity of cholesterol in phosphatidylcholine-cholesterol mixed model membranes are obtained by examining the orientational order parameter S of cholestane spin probe (CSL) that is obtained from electron spin resonance by spectral simulation. By introducing thermodynamic stability conditions of liquid mixtures, the variation of activity (or S) as a function of cholesterol mole fraction is utilized to predict the concentration at which the phase separation occurs. These results for DMPC and cholesterol binary mixtures agree very well with those of Tempo-partitioning experiments. The comparison of activity coefficients and the phase boundary in DMPC/cholesterol mixtures with those of POPC/cholesterol mixtures suggests that acyl chain unsaturation leads to poorer mixing of cholesterol in phosphatidylcholine model membranes at higher temperatures (i.e., greater than 35 degrees C). In ternary solutions of DMPC, POPC, and cholesterol, it is found that cholesterol shows less deviation from ideality than in either of the two binary mixtures, and this implies that the phase separation occurs at higher cholesterol concentration than in either of the two binary mixtures. The present analysis suggests that there may not be a critical point in DMPC/cholesterol mixtures, even though phase separation does occur. PMID- 2558734 TI - Conformation of spin-labeled melittin at membrane surfaces investigated by pulse saturation recovery and continuous wave power saturation electron paramagnetic resonance. AB - Melittin spin-labeled specifically with a nitroxide at positions 7, 21, 23, or the amino terminus was bound to phospholipid membranes, and the exposure of the spin label to the aqueous phase was investigated by measurement of Heisenberg exchange with chromium oxalate in the solution. The exchange frequency was determined by saturation recovery electron paramagnetic resonance (EPR) using a loop-gap resonator. This method allows use of very low concentrations (less than 1 mM) of chromium oxalate compared with conventional measurements of EPR line broadening (typically 50 mM), thus avoiding problems associated with high metal ion concentration. Differences in exchange frequency between the various positions were also estimated by continuous wave power saturation methods. In either approach, the spin label at lysine 7 was found to be the most exposed to chromium oxalate whereas that at lysine 23 was found to be the least exposed. This is consistent with a model for the membrane bound peptide in which an amphiphilic helix lies with its axis parallel to the bilayer surface and the hydrophobic moment points toward the bilayer interior. PMID- 2558735 TI - Guanosine-5'-O-(3-thiotriphosphate) modifies kinetics of voltage-dependent calcium current in chick sensory neurons. AB - Internal perfusion with the G-protein activator guanosine-5'-O-(3 thiotriphosphate) (GTP-gamma S) mimics the effect of noradrenaline and dopamine on the voltage-dependent calcium current in chick dorsal root ganglion (DRG) cells. With 100 microM GTP-gamma S in the pipette, the current at +10 mV was depressed by approximately 50%, with a 10-fold increase of its time to peak. The activation time course of the control calcium current could be approximated with a single exponential curve, whereas with GTP-gamma S the activation time course was double exponential, with time constants tau 1 and tau 2. 2 mM Mg-ATP in the pipette prevented the GTP-gamma S-induced current decrease in 70% of the cells, but the time course of the current was always double exponential. From -50 mV, the current at +10 mV was best fitted with tau 1 = 1.7 +/- 0.5 and tau 2 = 25.6 +/- 5.5 in seven cells. Both time constants decreased with increasing depolarizations. In the first 2 min of recording, the current changed with time. However, both tau 1 and tau 2 were constant, whereas the relative contribution of the slow component increased from 10 to 70%. In addition, the effect was independent of the holding potential in the range from -100 to -30 mV. These results suggest that the activation of a G-protein causes a fraction of the high threshold calcium channels to switch to a new closed state, with slower opening kinetics. PMID- 2558736 TI - Microcalorimetric studies of conformational transitions of ferricytochrome c in acidic solution. AB - The conformational transitions of ferricytochrome c in acidic solutions with different NaCl concentrations have been studied by scanning and isothermal microcalorimetry. It is shown that ferricytochrome c adopts three different forms which are realized under the considered conditions: native, denatured (unfolded) and a compact native-like form with unique tertiary structure. The thermodynamic parameters of the corresponding transitions have been measured and the changes in the number of bound ligands (H+ and Cl-), accompanying these transitions, have been determined by analyzing the temperature, pH and ionic strength dependence of these parameters. PMID- 2558738 TI - Export of proteins across membranes: the helix reversion hypothesis. AB - A model is presented which explains the biological role of the leader peptide in protein export. Along the lines of this model, the conformational changes of a protein with environment serves as a general mechanism for translocation. The leader peptide in the cytoplasm takes a hairpin like conformation which reverts to an extended helix upon integration into the membrane. The essential features of this model are in accord with recent results of protein export. PMID- 2558737 TI - A theoretical method for distinguishing between soluble and membrane proteins. AB - A method for distinguishing between membrane and soluble proteins in an amino acid sequence was developed, using only two parameters associated with the hydrophobicity: the average hydrophobicity and the power spectral density of period longer than 30 residues. The power spectral density was calculated by a maximum entropy method of Fourier transformation. Membrane proteins could be distinguished from soluble proteins with a distinction rate as high as 97%. This fact strongly suggests that the morphology of proteins, i.e., membrane or soluble forms, is determined thermodynamically through the hydrophobicity of polypeptides. PMID- 2558739 TI - [Stress-induced changes in the functioning of the Ca-transport system of the cardiac sarcoplasmic reticulum and its resistance to endogenous damaging factors]. AB - The kinetic characteristics of a calcium transport by sarcoplasmic reticulum was measured in homogenate of rat heart by means of Ca-selective electrode in the presence of oxalte and azide. As a function of the increasing CaCl2 concentration, the rate of Ca2+ uptake demonstrate a linear (in the physiological concentration field) and saturable components. Prolonged immobilization was accompanied by a decrease in Ca-concentration dependence of Ca2+ uptake. Nonsignificant fall of Vm was accompanied by simultaneous decrease of KM by 30% after stress. Storage of heart homogenates revealed three-fold decrease of inactivation rate in the stress series. Initial concentration of free Ca2+ and its raise during storage was increased in heart homogenates, prepared after stress. The role of calcium pump resistance to endogenous damage factors in the stress-induced heart disturbance are discussed. PMID- 2558740 TI - [A comparative evaluation of the level of sphingomyelinase activity in liver cell nuclei and in the brain]. AB - It was discovered that there is sphingomyelinase activity in the rat liver nuclei. The maximum of enzyme activity is at pH 7.1. The data obtained demonstrated that the main part of sphingomyelinase is located in the nuclear membrane. Comparison of sphingomyelinase activity in cell nuclei, liver and brain homogenates shows high level of enzyme activity in the nuclei. The authors discuss possible participation of sphingomyelinases in changes of phospholipids composition in nuclear structure under different functional activity of cell nuclei. PMID- 2558741 TI - [The action of emoksipin on the basal activity of cyclic nucleotide phosphodiesterase and on the late receptor potential of the isolated retina]. AB - The influence of emoxypin (derivate of 3-hydroxypyridine) upon the late receptor potential (LRP) and activity of the cyclic 3',5'-nucleotide phosphodiesterase (PDE) have been investigated. The inhibition of PDE and increase of the amplitude of LPR have been shown. The curve (RP as a function of the stimulus light intensity) was moved towards the lesser lighting and the time of the achievement of the maximum was increased. Thus, emoxypin produces an effect on the LRP like classical inhibitors of PDE. It is suggested that increase of the functional activity of the retinae upon the influence of emoxypin in caused by the influence of the one towards the system of the cyclic nucleotides. PMID- 2558742 TI - [Changes in pain reactions and 3H-naloxone binding to opiate receptors of the hypothalamus and midbrain in rats after repeated swimming in cold water]. AB - The dynamics of nociceptive reactions and character of 3H-naloxone binding to hypothalamus and midbrain synaptic membranes were studied in rats subjected to repeated cold swim stress (3 min. daily during 3, 5 and 15 days). It was shown that an increase of latencies of background nociceptive reactions (hot-plate and tail-flick tests) was accompanied by an ambiguous changes of kinetic parameters of 3H-naloxone binding in the studied brain structures. The results suggest that an increase of antinociceptive systems tone under repeated cold swim stress may be caused by a dynamic transformation of opiate u-receptor apparatus in various brain structures. PMID- 2558743 TI - [Species and functional differences in NMDA receptors]. AB - N-methyl-DL-aspartate induced in mice and rats hyperactivity and convulsions. In the mice convulsions effectively suppressed by selected antagonists of NMDA receptors AP 5 and AP 7, and also diazepam and ketamine. But hyperactivity very little attenuated by these substances in high doses. In the rats all these substances on the contrary suppressed hyperactivity and convulsions in the same degree. In the rats hynurenic acid prevented hyperactivity and convulsions. Observations point out to the fact that in mice there are two pharmacological and functional types of NMDA receptors whereas in rats there is only one type which is different from NMDA mice receptors. PMID- 2558745 TI - [The effect of morphine and opiate receptor agonists on arterial pressure and the bioelectrical activity of the renal nerve during stimulation of the midbrain central gray substance]. AB - It has been demonstrated in experiments on unrestrained and unanesthetized curarized cats that periaqueductal gray matter stimulation produce sympathetic activating action, raise arterial pressure and heart rate, but at the same time is not effective enough to suppress the nociceptive shifts of haemodynamic reactions. Opioid mechanisms of spinal cord plays an essential role in sympathetic-activating action of periaqueductal gray matter. It is suggested that the influence of antinociceptive areas of the brain stem on sympathetic haemodynamic regulation is one of the causes of resistance of nociceptive haemodynamic reactions to narcotic analgetics. PMID- 2558744 TI - [Adaptive changes in sigma and phencyclidine receptors during the long-term use of haloperidol and raclopride in rats]. AB - The experiments on male albino rats have shown that 15 days haloperidol (0.5 mg/kg) and raclopride (1 mg/kg) treatment, but not acute administration, causes the increase of density of sigma receptors in the brain. The number of phencyclidine receptors was also elevated, but this increase was not statistically evident. The behavioral effects of ketamine (5 mg/kg) were evidently decreased after long-term haloperidol and raclopride treatment. The motor stimulation and stereotyped behavior induced by apomorphine (0.15 mg/kg) were increased only after treatment of haloperidol, but not raclopride. It seems probable that repeated neuroleptic (haloperidol and raclopride) treatment causes the hyposensitivity of sigma and phencyclidine receptors, despite the increase of their number. It is possible that this change is related to the depolarization inactivation of dopamine neurons caused by repeated neuroleptic administration. PMID- 2558747 TI - Familial colorectal cancer and familial adenomatous polyposis. AB - Familial adenomatous polyposis (FAP) affects around 1 in 10,000 individuals; the gene for this condition was recently shown to be located on chromosome 5, and it is only a matter of time before its precise location and function are determined, making prephenotypic, and even prenatal, diagnosis more generally available and reliable. In the mean time, care of FAP families will continue to depend on careful registration of family information, prophylactic bowel surgery and surveillance for other potentially serious manifestations of the disease. Upper gastrointestinal malignancies and desmoid tumours have overtaken colorectal cancer as the leading causes of death in some centres. Other dominantly-inherited colorectal cancer syndromes produce less striking phenotypes, but affect far more individuals than FAP. It appears that there are two patterns of hereditary non polyposis colorectal cancer (HNPCC) syndromes, one involving cases of bowel cancer alone, the other associated with breast and gynaecological cancers; these may prove to be variable expressions of a common gene abnormality. More effort is required by clinicians managing cases of colorectal cancer to identify affected families in order to offer surveillance and appropriate treatment in the hope that such measures may prevent cancer in family members. PMID- 2558748 TI - Isotope dilution assay for urinary methanesulfinic and methanesulfonic acids: application to detection of methylthio turnover. AB - Methanesulfinic and methanesulfonic acids were shown to be present in urine from rats fed laboratory rat diet. The daily excretions of each acid were equivalent at 0.47 mumol day-1. The excretion of methanesulfinic acid increased about ninefold and that of methanesulfonic doubled when rats were dosed with 17 mumol of pentachlorothioanisole. The excretion of methanesulfinic acid above control levels in urine may be a method for detecting methylthio displacements that occur during the intermediary metabolism of xenobiotics. PMID- 2558746 TI - [The effect of the stimulation of opioid receptors on lymphocyte functional activity in vivo]. AB - With the aid of lymphocytes' spontaneous adhesion test and the reaction of blast transformation with phytohemagglutinin (PHA) the influence of met-enkephaline upon the functional activity of lymphocytes of healthy donors was studied in vitro. It was shown, that met-enkephaline stimulate lymphocyte spontaneous adhesion and the lymphocyte PHA-induced proliferative activity. The stimulating effects of met-enkephalin revealed in vitro were blocked by the opioid receptor blocker naloxone. PMID- 2558750 TI - Antiestrogen and antiandrogen administration reduce bone mass in the rat. AB - In mature female and male rats sex hormone deficiency was produced by surgical castration and by antiestrogen or antiandrogen administration. For the latter purpose we used the nonsteroidal antiestrogens tamoxifen, keoxifene (LY156758) and tetramethylhexestrol, and the steroidal antiandrogen cyproterone acetate. Dosages of 0.4 mg tamoxifen/rat/day and isomolar dosages of keoxifene and tetramethylhexestrol led to a bone mass reduction which was comparable to ovariectomized rats. Cyproterone acetate showed, at 10 mg/rat/day, a similar decrease in bone mass like orchidectomy. The often discussed intrinsic estrogen activity of the antiestrogens was present only in the highest dosage tested of tamoxifen. Keoxifene and tetramethylhexestrol showed no estrogenic effects, but this may be a dosage problem. Cyproterone acetate revealed no androgenic side effects. These results indicate that antigonadal hormone drugs reduce bone mass to a varying extent. PMID- 2558749 TI - Calcium modulation of the parathyroid hormone-sensitive adenylate cyclase in ROS 17/2.8 cells: effects of N-(6-aminohexyl-5-Cl-naphthalene sulfonamide) (W-7) and trifluoperazine (TFP). AB - The calcium modulation of the cyclic 3',5'-adenosine monophosphate (cAMP) response to parathyroid hormone (PTH) was studied in a clonal osteosarcoma cell line ROS 17/2.8. CaCl2 was found to stimulate the PTH-sensitive cAMP response of intact cells. At the maximal concentration of 1 mM CaCl2, the maximum response to PTH was increased, but the ED50 for PTH and the time course of maximal cAMP production were not affected. Verapamil blunted, while the cation ionophore A23187 enhanced, the stimulatory effect of CaCl2. Trifluoperazine (TFP) and N-(6 aminohexyl-5-Cl-naphthalene sulfonamide) (W-7) inhibited the stimulatory effect of CaCl2. In membranes prepared in the presence of 0.1 mM CaCl2, a biphasic effect of CaCl2 was demonstrated: stimulation at concentrations of 60-100 microM, and an inhibition above 200 microM, when adenylate cyclase was assayed in the presence of 200 microM EGTA. Addition of exogenous calmodulin to membranes prepared in the presence of EGTA did not have any effect on the PTH-sensitive adenylate cyclase activity, suggesting that endogenous calmodulin was not effectively stripped from the membranes by EGTA treatment. It is concluded that Ca2+ has both a stimulatory and an inhibitory role in modulating PTH-sensitive adenylate cyclase in ROS 17/2.8 cells by as yet unknown mechanisms, and that the involvement of endogenous calmodulin is implicated. PMID- 2558751 TI - Prostaglandin E2 initially inhibits and then stimulates bone resorption in isolated rabbit osteoclast cultures. AB - Osteoclasts were isolated from the long bones of neonatal rabbits and cultured on devitalized bovine bone slices for 8, 24, 48 and 72 h with and without prostaglandin E2 (PGE2) (10(-6) M). The number of osteoclasts present at the end of the culture periods was counted after staining the cells for tartrate resistant acid phosphatase (TRAP). After removal of the cells, the resorption lacunae excavated by the osteoclasts were observed by scanning electron microscopy (SEM) and their size and depth calculated by computer-assisted morphometric and stereomorphometric techniques. PGE2 had no effect on the number of TRAP positive multinucleated osteoclasts, but decreased the number of TRAP positive mononuclear cells. The total area of the excavated pits and the area excavated per osteoclast in PGE2-treated cultures were decreased by 62 and 58% respectively after 8 h in culture. After 24 h in culture, the total excavated area and the excavated area per osteoclast were still 44 and 38% lower in the PGE2-treated cultures than in the corresponding control cultures. However, after 48 h of culture, resorptive activity in PGE2-treated cultures was consistently greater than in control cultures. In the course of a 48 h culture period, the PGE2 concentration decreased from 1.0 x 10(-6) to 0.3 x 10(-6) M. Thus, despite the continuous presence of PGE2, the resorptive activity of osteoclasts not only recovered from the transient inhibitory effect of PGE2, but was actually greater than in the control cultures. This confirms that the effects of PGE2 in isolated osteoclast preparations are inhibitory in short term cultures, but shows that the effects of PGE2 in such preparations are stimulatory in longer term cultures. Proliferating stromal cells with osteoblast-like characteristics comprised approximately 45% of the 'osteoclast' cultures at the start of the cultures, but their number increased to 93% of the total cell population at 48 h and to 98% at 72 h. Our results suggest that the PGE2-induced stimulation of osteoclastic activity represents an indirect effect mediated by stromal cells derived from bone marrow. Our results also indicate that the increased resorptive activity in PGE2 treated cultures can be accounted for by an increase in the size of the resorption lacunae and is not caused by an increase in osteoclast number. PMID- 2558752 TI - Rheumatology in Rasigueres, 1989. PMID- 2558753 TI - Epidemiologic survey of bovine diseases in Suriname. AB - A seroepidemiologic survey of cattle diseases was undertaken in Suriname in 1985 to help assess the livestock disease situation in that country. The six diseases covered by the survey were bovine coronavirus infection, bovine rhinotracheitis, bovine virus diarrhea, brucellosis, parainfluenza-3 infection, and respiratory syncytial virus infection. The results indicated relatively low prevalences of these diseases compared to the prevalences found in most developed countries. The reasons for this are uncertain, but the finding suggests that the cattle population in Suriname could lack extensive exposure to these diseases and so could be highly susceptible to them. In addition, the evident need for more thoroughgoing survey data points up the need to establish a continuous animal data health monitoring system in Suriname--as well as in other developing countries where there is a need to objectively assess the livestock disease picture. PMID- 2558754 TI - The value of nutrition surveillance: a case study from the United States. PMID- 2558755 TI - Ubiquitin: new insights into chronic degenerative diseases. AB - Ubiquitin is a 76-amino-acid protein and is perhaps the most conserved gene product in evolution. It modulates degradation of abnormal or damaged proteins and belongs to the class of heat-shock proteins induced in conditions of cell stress. Recent work shows that ubiquitin is involved in several chronic degenerative diseases characterized by the formation of cellular inclusion bodies. The ubiquitin response to cell injury appears to be cytoprotective and particularly important in diseases of the nervous system. PMID- 2558756 TI - An in vitro study of the relationship between GABA receptor function and propulsive motility in the distal colon of the rabbit. AB - 1. The effects of gamma-aminobutyric acid (GABA), 3-aminopropane sulphonic acid (3-APS) and baclofen on spontaneous, electrically-induced and propulsive motility were investigated in rabbit distal colon. 2. In unstimulated longitudinal (LMPs) and circular muscle strip preparations (CMPs) 3-APS (10-200 microM) and GABA caused a clear-cut relaxation susceptible to desensitization. Baclofen (10-200 microM) caused relaxation in a minority (30%) of preparations. The 3-APS response was sensitive to tetrodotoxin (TTX; 1 microM), SR 95531 (a novel competitive GABAA-receptor antagonist) (10 microM), picrotoxinin (30 microM), and insensitive to hyoscine (1 microM) and to a combination of prazosin (1 microM) and propranolol (1 microM). The baclofen response was antagonized by 5-aminovaleric acid (DAVA, 500 microM), TTX and hyoscine and resistant to GABAA-receptor and adrenoceptor blockade. GABAA-receptors were therefore associated with non adrenergic non-cholinergic (NANC) inhibitory nerve activation while GABAB receptors were involved in depression of cholinergic tone of smooth muscle. GABA (10-200 microM) elicited both above mentioned effects. 3. In LMPs, baclofen (10 200 microM) dose-dependently inhibited submaximal responses to both cholinergic and NANC inhibitory nerve stimulation. This effect was resistant to SR 95531 and picrotoxinin and prevented by DAVA and baclofen desensitization. GABA (10-200 microM) mimicked the action of baclofen. GABA inhibitory effects persisted in the presence of GABAA-receptor blockade. 4. In segments of distal colon, GABA and baclofen (1-200 microM), but not 3-APS (1-200 microM), dose-dependently decreased the velocity of propulsion of an intraluminally-distended balloon. This effect was antagonized by DAVA and GABA or baclofen desensitization and resistant to SR 95531 and picrotoxinin. These antagonists per se had no effect on propulsion. In preparations in which propulsion was slowed by hyoscine (1 microM), baclofen caused no consistent further depression of propulsive activity. 5. Our results show that GABAA- and GABAB-receptors are present in rabbit colon. GABAA-receptor stimulation activates NANC inhibitory nerves without apparently affecting propulsion. GABAB-receptors are associated with a reduction of neural (mainly cholinergic) activity subserving muscular tone and peristalsis and appear to be located on both cholinergic and NANC inhibitory nerves. However, the persisting propulsive activity during suppression of GABAA- and GABAB-receptor function suggests that GABA in enteric neurones is not crucial for the neural circuitry subserving colonic peristalsis in this species. PMID- 2558758 TI - Studies on the adrenomedullary dependence of kappa-opioid agonist-induced diuresis in conscious rats. AB - 1. The dependence of kappa-opioid agonist-induced diuresis, upon an intact and functional adrenal medulla in conscious rats, was investigated in order to test the hypothesis that the diuresis is mediated by a blood-borne 'diuretic factor', of adrenomedullary origin, released by kappa-opioid receptor stimulation. 2. Confirming previous observations, adrenal demedullation significantly attenuated diuretic responses to the kappa-opioid agonists U50488H, ethylketocyclazocine (EKC) and tifluadom, but did not affect basal urine output, furosemide-induced diuresis or the antidiuretic response to the mu-opioid agonist, buprenorphine. Naloxone abolished U50488H-induced diuresis, confirming an involvement of opioid receptors. 3. Transfusion studies established that blood, from intact rats treated with U50488H, induced diuresis in intact and demedullated recipient rats, whether or not the recipients had been pretreated with naloxone. However, blood from demedullated rats treated with U50448H was unable to induce diuresis when administered to intact or demedullated recipients. 4. It is concluded that kappa opioid agonist-induced diuresis is dependent upon an intact and functional adrenal medulla and appears to be mediated by a blood-borne 'diuretic factor' of adrenomedullary origin. PMID- 2558757 TI - [3H]-idazoxan binds with high affinity to two sites on hamster adipocytes: an alpha 2-adrenoceptor and a non-adrenoceptor site. AB - 1. [3H]-idazoxan labels a single population of high affinity sites (Kd 2.26 +/- 0.02 nM; Bmax 372 +/- 25 fmol mg-1 protein) in hamster adipocyte membranes. In the presence of 1 microM yohimbine to preclude binding to alpha 2-adrenoceptors, the density of [3H]-idazoxan binding sites was reduced (287 +/- 18 fmol mg-1 protein) without an apparent decrease in the affinity (Kd 2.19 +/- 0.24 nM) of the radioligand. 2. Displacement studies indicate that alpha-adrenoceptor ligands with an imidazoline side chain completely inhibit [3H]-idazoxan binding to hamster adipocyte membranes; in contrast, the alpha 2-adrenoceptor antagonists yohimbine, rauwolscine, BDF 6143 and phentolamine inhibited only 20-30% of the specific binding with affinity values consistent with an interaction at alpha 2 adrenoceptors. 3. The low potency of noradrenaline and adrenaline in displacing [3H]-idazoxan binding to the second site on hamster adipocyte membranes indicates that it is unlikely that this site is a type of adrenoceptor. 4. These results suggest that [3H]-idazoxan binds with high affinity to two sites in hamster adipocytes: an alpha 2-adrenoceptor and a non-adrenoceptor imidazoline site. PMID- 2558759 TI - Modulation of sodium current kinetics by chlorpromazine in freshly-isolated striatal neurones of the adult guinea-pig. AB - 1. The neurones of the striatum were freshly dissociated from the adult guinea pig brain by enzymatic and mechanical treatments. Sodium channel current kinetics in these neurones were measured using a whole cell variation of the patch-clamp technique. 2. Chlorpromazine, a neuroleptic, in micromolar concentrations reversibly reduced the amplitude of the sodium currents. Activation and inactivation time constants were not affected. The inhibition followed one-to-one binding stoichiometry. 3. The concentration-response curve shifted to the left when the holding potential was less negative. The EC50 shifted from 4.8 microM to 0.9 microM when the holding potential was changed from -120 mV to -70 mV. 4. The steady-state activation curve of the sodium current was not affected by chlorpromazine, whereas the steady-state inactivation curve was shifted in the negative direction. Consequently, the window current which is normally present at a potential range around -50 mV was decreased in the presence of chlorpromazine. 5. Successive sodium currents evoked by a train of depolarizing pulses (30 ms duration) to -10 mV showed a cumulative decrease in size during the application of chlorpromazine. However, such 'use-dependent' block was not observed when the pulse duration was reduced to 1 ms. 6. The recovery from inactivation in the presence of chlorpromazine, was expressed as a second order process. The faster component was similar to the recovery time course of the normal sodium channels. The slower component accounted for the use-dependent effect of chlorpromazine. 7. The results indicate that chlorpromazine binds to the resting sodium channels producing steady-state block at a very negative holding potential. When the membrane is depolarized, chlorpromazine binds to the inactivated form of the sodium channels with much higher affinity and stabilizes them in the inactivated state, slowing their kinetics. PMID- 2558760 TI - Muscarinic receptors coupled to phosphoinositide hydrolysis and elevated cytosolic calcium in a human neuroblastoma cell line SK-N-SH. AB - 1. The effects of the muscarinic agonist carbachol on phosphoinositide metabolism and its relationship to alteration of intracellular calcium were examined in SK-N SH human neuroblastoma cells. Muscarinic receptors on these cells are coupled to phospholipase C and the myo [2-3H]-inositol phosphates resulting from receptor activation of cells labelled with [3H]-inositol accumulate rapidly. The breakdown of both inositol monophosphate (InsP1) and inositol bisphosphate (InsP2) is sensitive to lithium with inhibition of the latter only observed at higher concentrations of this ion. 2. Use of the calcium indicator dye Fura 2 revealed that carbachol stimulates a biphasic increase in intracellular calcium. 3. Carbachol was able to stimulate both [3H]-inositol phosphate production and intracellular calcium levels with respective EC50 values of 15.9 +/- 1.0 microM and 10.7 +/- 3.2 microM, indicating that no amplification occurs between these steps in the signal transduction pathway. 4. Inositol 1,4,5 trisphosphate (Ins(1,4,5)P3) released 45Ca2+ in a stereospecific and dose-related manner from intracellular stores of permeabilised cells. 5. These results suggest that this cell line may represent a useful model system to investigate receptor-mediated phosphoinositide metabolism and calcium homeostasis. PMID- 2558761 TI - A phorbol diester-induced enhancement of synaptic transmission in olfactory cortex. AB - 1. Extracellular field synaptic potentials were recorded from pial surface slices of guinea-pig olfactory cortex maintained in vitro. 2. Phorbol 12,13-dibutyrate (0.1-10 microM) enhanced the amplitude of the evoked potential (by 51.2 +/- 10.4% with 1 microM) in normal solution. When the evoked potential was partially depressed by Cd, Co, Mn or a reduced Ca concentration, phorbol 12,13-dibutyrate (1 microM) induced a much larger enhancement of the evoked potential (196.5 +/- 24.4% increase). Phorbol 12,13-diacetate and mezerein had similar effects but were less potent. 4 beta-Phorbol (10 microM) had no effect. 3. The diacylglycerol analogues, dioctanoylglycerol (100-1000 microM), 1-oleoyl-2-acetylglycerol (100 500 microM) or diolein (100 microM) had no effect on the evoked potentials, either alone or in the presence of Cd. 4. The isoquinolinylsulphonamide inhibitor (H-7) of protein kinase C slightly enhanced the e.p.s.p. and had no effect on the potentiation produced by phorbol ester. Another protein kinase C inhibitor, acridine orange (100-1000 microM), had no effect on the action of phorbol ester. 5. These results show that transmitter release, as at other synapses, is enhanced by phorbol esters but Ca did not potentiate this action. The pharmacological profile of the effect on transmitter release differed from that of protein kinase C in cell-free preparations and therefore it is unclear whether protein kinase C was involved in the present study. PMID- 2558764 TI - Evidence for pre-junctional muscarinic receptors on pulmonary sympathetic nerves in the guinea-pig. PMID- 2558762 TI - Temporal changes in the calcium-dependence of the histamine H1-receptor stimulation of cyclic AMP accumulation in guinea-pig cerebral cortex. AB - 1. 2-Chloroadenosine (2CA) causes a maintained rise in adenosine 3':5'-cyclic monophosphate (cyclic AMP) content of guinea-pig cerebral cortical slices which is augmented by addition of histamine. We have investigated the temporal profile of the sensitivity of this response to calcium. 2. Rapid removal of extracellular calcium with EGTA (5 mM) at 2CA (30 microM)-induced steady state caused a slight increase in the cyclic AMP response to 2CA alone and completely abolished the augmentation produced by histamine (0.1 mM) added 20 min later. When EGTA was added only 2 min before histamine, the augmentation was reduced by 72%. 3. The calcium sensitivity of the histamine response was also indicated in studies in which EGTA was added 1 or 3 min after histamine at 2CA-induced steady state. Following addition of EGTA at either of these times, the augmentation was not maintained. 4. When calcium was rapidly removed with EGTA once a steady state level of cyclic AMP had been achieved with histamine, the augmentation response was maintained. This was despite the fact that EGTA had a similar effect on both extracellular free calcium and tissue calcium content when it was applied before or after histamine. 5. The 2CA response was augmented by phorbol esters (which mimic the actions of diacylglycerol) in a calcium-independent manner. 6. These results suggest that calcium is important for the initiation and early stages of the histamine-induced augmentation response. The apparent lack of calcium sensitivity of the response at later stages could mean that calcium is not involved in the maintenance of the response or that the intracellular machinery involved in the augmentation process becomes more sensitive to calcium as the response progresses, such that it becomes able to operate at a much lower level of intracellular calcium. A possible role for diacylglycerol in the maintenance of the response is discussed. PMID- 2558765 TI - The effect of sensory nerve depletion on cholinergic neurotransmission in guinea pig airways. PMID- 2558763 TI - Pharmacological profile of leukotrienes E4, N-acetyl E4 and of four of their novel omega- and beta-oxidative metabolites in airways of guinea-pig and man in vitro. AB - 1, The biological effects of metabolites of leukotriene E4 (LTE4) i.e. N-acetyl LTE4 (N-AcLTE4), 20-COOH-LTE4, 20-COOH-N-AcLTE4, as well as 18-COOH-19,20-dinor LTE4 (dinor-LTE4) and 16-COOH-17,18,19,20-tetranor-14,15-dihydro-LTE4 (tetranor LTE4) were investigated on superfused strips of guinea-pig trachea (GPT) and lung parenchyma (GPP) in vitro. 2. The actions of LTE4 were studied in isolated, superfused strips of human lung parenchyma (HP) and bronchus (HBr), in comparison with LTD4 and histamine. Effects of N-AcLTE4, the 20-carboxy metabolites, dinor LTE4 and tetranor-LTE4 were also investigated in HBr. 3. N-AcLTE4 (0.1-10 nmol) induced dose-related contractions of GPT and was approximately 100 times less active than LTD4 (3-100 pmol). 4. In GPP, N-AcLTE4 (0.01-3 nmol) was equiactive with LTE4 (0.01-1 nmol) and approximately one order of magnitude less active than LTD4 (1-300 pmol). Contractions caused by N-AcLTE4 and LTE4 were very similar and approximately twice as sustained as those due to LTD4. 5. LTE4 (0.1-30 nmol) contracted strips of HP and HBr and was about 2-3 orders of magnitude less active than LTD4. As in GPP, the effect of LTE4 was more protracted than that of LTD4. Actions of N-AcLTE4 were similar to those of LTE4 in HBr. 6. 20-carboxy-LTE4, 20 carboxy-N-AcLTE4, dinor-LTE4 and tetranor-LTE4, all at 0.3-30 nmol, were inactive in GPT, GPP and HBr. 7. Indomethacin (2.8 microM) potentiated the effect of N AcLTE4 in GPT, inhibited its contraction in GPP but did not affect that due to LTE4 in either HP or HBr. FPL 55712 (1.9 microM) antagonised leukotriene-induced contractions in GPT, GPP and HBr. 8. In conclusion, the metabolism of LTD4 to LTE4 or N-AcLTE4 may represent a detoxification but not an inactivation of cysteinyl-containing leukotrienes, since both metabolites still retain considerable biological activity in guinea-pig and human airways in vitro. However, further metabolism of LTE4 and N-AcLTE4 appears to result in inactivation of leukotrienes. PMID- 2558766 TI - Lower cortical beta-adrenoceptor binding sites in post-mortem samples from depressed suicide victims. PMID- 2558767 TI - Modulation of inositol (1,4,5)-trisphosphate concentration by cyclic AMP in rat cerebral cortex slices. PMID- 2558768 TI - Involvement of excitatory amino acid receptors in epileptiform activity in the rat entorhinal cortex and dentate gyrus in vitro. PMID- 2558769 TI - Are changes in glutamatergic mechanisms responsible for kindling to the benzodiazepine receptor inverse agonist, FG 7142? PMID- 2558770 TI - Effects of the calpain inhibitor E64-d on platelet activation responses. PMID- 2558771 TI - Consistent reduction of ACTH responses to stimulation with CRH, vasopressin and hypoglycaemia in patients with major depression. AB - Eleven patients with major depression and 12 control subjects were administered corticotropin-releasing hormone (CRH), aqueous arginine vasopressin (AVP), and insulin hypoglycaemia (IH) to test for differences in hypothalamic-pituitary adrenal (HPA) axis function. Patients with major depression demonstrated lower ACTH responses to CRH when compared with controls, and a trend toward such after administration of AVP. Despite lower ACTH responses in patients with depression, there were no differences in cortisol responses to these stimuli. In the CRH and AVP tests, there was no correlation between the basal cortisol and ACTH responses in either controls or patients, but in the IH test there was a negative correlation between these responses for both groups. The ACTH responses to CRH and AVP were positively correlated in controls and patients. Cortisol responses to all three provocative stimuli were positively correlated in both subject groups. These findings are consistent with the hypothesis that hypothalamic or supra-hypothalamic overactivity may be involved in the development of HPA-axis abnormalities in patients with depression. PMID- 2558772 TI - The measurement of transmembrane cation transport in vivo in acute manic illness. AB - We have used a novel technique to assess the transport of cations across the erythrocyte membrane in vivo in unmedicated patients suffering an acute manic illness. The results show that erythrocyte cation transport via the sodium-pump enzyme Na+,K+-ATPase is increased in manic patients compared with healthy controls. PMID- 2558773 TI - The effect of lithium on cation transport measured in vivo in patients suffering from bipolar affective illness. AB - We have investigated cation transport in vivo in patients being treated with lithium for bipolar affective illness by studying the disposition of rubidium after an oral load of rubidium chloride. The rate of erythrocyte cation transport was increased in the patients when compared with matched healthy volunteers. However, the rate of in-vivo erythrocyte rubidium accumulation in the euthymic treated patients was significantly lower than in a matched group of unmedicated manic patients. The regulation of specific pathways for cation transport may be altered in individuals predisposed to affective illness. PMID- 2558774 TI - Paraneoplastic hypercalcaemia in association with schistosomal bladder cancer. PMID- 2558775 TI - A comparison of the anticonvulsant effects of competitive and non-competitive antagonists of the N-methyl-D-aspartate receptor. AB - The anticonvulsant activity of two competitive antagonists of the N-methyl-D aspartate (NMDA) receptor, 2-amino-7-phosphonoheptanoic acid (APH) and 3-[2 carboxypiperazin-4-yl]-propyl-1-phosphonate (CPP), and two non-competitive NMDA antagonists, phencyclidine (PCP) and (+)-5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine (MK-801), were compared in 4 models of induced seizures in mice. All 4 drugs protected against tonic extensor seizures induced by pentylenetetrazol (PTZ), by submaximal (15 mA) electroconvulsive shock (ECS) and by maximal (50 mA) ECS. Similar orders of potency (i.e., MK-801 greater than PCP greater than or equal to CPP greater than APH) were seen in each of the 3 seizure models. All 4 drugs failed to block clonic seizures induced by picrotoxin in the dose ranges that protected from tonic seizures. These data are consistent with other data demonstrating that competitive and non-competitive NMDA antagonists have similar pharmacologic effects. These results also support the suggestion that the anticonvulsant effects of competitive and non-competitive NMDA antagonists are mediated by the NMDA receptor-ionophore complex. PMID- 2558776 TI - Regulation by estradiol of GABAA and GABAB binding sites in the diencephalon of the rat: an autoradiographic study. AB - Using in vitro quantitative autoradiography we studied the in vivo effects of estradiol on GABAA and GABAB receptors in the rat brain. In all the areas studied (suprachiasmatic nucleus, medial preoptic area, striatum, frontal cortex), estradiol failed to significantly affect the GABAA receptor density. Chronic treatment with estradiol led however in the suprachiasmatic nucleus and the striatum to a decrease in the density of GABAB receptors. GABAB receptor regulation by estradiol was found to be area-specific within the hypothalamus since it was not observed in the medial preoptic area. The down regulation of GABAB receptors in the suprachiasmatic nucleus induced by estradiol treatment might thus explain the inhibitory effect of the steroid on the GABA control of serotonin metabolism we recently reported. PMID- 2558777 TI - Non-N-methyl-D-aspartate receptors may mediate ipsilateral excitation at lateral superior olivary synapses. AB - Principal cells of the lateral superior olivary nucleus (LSO) are thought to receive a direct excitatory input from spherical bushy cells located in the ipsilateral ventral cochlear nucleus (VCN) and an indirect input from the contralateral VCN globular bushy cells via a secure synapse in the medial nucleus of the trapezoid body (MNTB). MNTB bushy cells project to the somata and proximal dendrites of LSO principal cells. LSO neurons display phasic 'chopper' temporal response patterns to ipsilateral tone-burst stimuli at characteristic frequency (CF), while binaural stimuli suppress this ipsilaterally evoked activity. This suppression is sensitive to interaural differences in intensity, phase and time, suggesting a role for these neurons in the localization of sound in space. In the present study, the nature of the neurotransmitter mediating fast ipsilateral excitation of LSO neurons was examined using iontophoretic application of excitant amino acid (EAA) agonists and antagonists. N-methyl-D-aspartate (NMDA) and quisqualate (QUIS) were used as agonists, while the selective NMDA receptor antagonist D. L-2-amino-5-phosphonovaleric acid (APV), and the non-selective receptor EAA antagonist cis-2,3-piperidine-dicarboxylic acid (PDA) were used to study ipsilaterally evoked neuronal responses. In 3 additional experiments the selective non-NMDA receptor antagonist 6,7-dinitroquinoxaline-2,3-dione (DNQX) replaced PDA. Ipsilateral, tone-evoked and spontaneous activities were generally enhanced by EAA agonists while partial blockade of tone-evoked, ipsilateral excitation was observed with EAA antagonists. Both PDA and DNQX more effectively blocked ipsilateral tone-evoked excitations and spontaneous activity than did the NMDA-receptor antagonist, APV.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558778 TI - Activation by high intensity peripheral nerve stimulation of adrenergic and opioidergic inhibition of a spinal reflex in the decerebrated rabbit. AB - The short-latency sural to gastrocnemius reflex in the decerebrated rabbit was depressed for 20-30 min following high intensity conditioning stimulation of the common peroneal nerve. This effect was observed in animals with or without spinal section, but was greater in non-spinalized preparations. Graded conditioning stimuli showed that it was necessary to activate fine myelinated common peroneal axons to inhibit the reflex. In spinalized rabbits, maximal inhibition was achieved with conditioning stimulation of fine myelinated axons and was completely reversed by the opioid antagonist naloxone. In non-spinalized rabbits, maximal inhibition was only obtained with conditioning stimuli which activated non-myelinated axons. In these preparations the effects of common peroneal nerve stimuli were only blocked by co-administration of naloxone with the alpha 2 adrenoceptor antagonist idazoxan. Thus high intensity peripheral nerve stimuli activated a segmental opioidergic and a supraspinal adrenergic suppression of the sural-gastrocnemius withdrawal reflex. Such long-lasting suppression of reflex excitability may contribute to recovery from intensely noxious stimuli. PMID- 2558779 TI - Characterization of opioid receptors in rat nucleus accumbens following mesolimbic dopaminergic lesions. AB - The present study investigated the cellular localization of mu, delta and kappa opioid receptors in the rat nucleus accumbens in relation to dopaminergic neurons. Dopaminergic terminals were destroyed by intra-accumbens injections of the neurotoxin 6-hydroxydopamine (6-OHDA). Fourteen days after dopaminergic denervation, receptor binding assays and quantitative in vitro autoradiography with highly selective radioligands demonstrated that the density of mu opioid receptors in the nucleus accumbens was decreased by 30 +/- 6%. There was no change in delta or kappa receptors in the accumbens, a finding which indicates that the loss of mu opioid receptors was specific. A time course study demonstrated that the loss of mu receptors lagged behind the depletion of dopamine by about 5 days. Destruction of intrinsic neuronal cell bodies and dendrites by injection of ibotenic acid into the accumbens resulted in a loss of 36 +/- 3% of mu opioid receptors. Co-injection of 6-OHDA and ibotenic acid decreased mu receptors by 41 +/- 4%, only slightly more than the loss caused by ibotenic acid alone. These results suggest that only a small number of mu opioid receptors in the nucleus accumbens are located on dopaminergic terminals and are consistent with the possibility that the loss of opioid receptors following denervation of dopaminergic fibers in the accumbens is the result of transsynaptic degeneration. PMID- 2558780 TI - Melatonin inhibits cyclic AMP and cyclic GMP accumulation in the rat pituitary. AB - Subnanomolar concentrations of melatonin inhibit cyclic AMP and cyclic GMP accumulation in neonatal rat anterior pituitary stimulated in vitro with luteinizing-hormone releasing-hormone. Melatonin also inhibited forskolin stimulated cyclic AMP accumulation in pars tuberalis. Inhibition of cyclic AMP accumulation is specific for melatonin, since its analogs N-acetylserotonin and 5 methoxytryptamine are 1000 times less potent. Cyclic nucleotides may thus serve as second messengers transducing the effect of melatonin on cellular level. PMID- 2558781 TI - Nerve growth factor (NGF) reverses axotomy-induced decreases in choline acetyltransferase, NGF receptor and size of medial septum cholinergic neurons. AB - Intraventricular nerve growth factor (NGF) infusion in the adult rat can prevent and also, if delayed, reverse the disappearance of most of the axotomized medial septum cholinergic neurons immunostained for choline acetyltransferase (ChAT). We have utilized the delayed NGF treatment protocol to (i) extend to 3 months the delay time between axotomy and NGF treatment, (ii) define the time course of their recovery, (iii) determine that immunostaining for the (lower affinity) NGF receptor (NGFR) parallels loss and reversal of the ChAT marker, and (iv) evaluate changes in cholinergic somal size following axotomy and subsequent NGF treatment. While NGF treatments starting only 7 days after the fimbria-fornix transection (axotomy) almost entirely restored the number of both ChAT- and NGFR-positive medial septum neurons, longer delayed (2-3 weeks) treatment brought about recovery from the baseline of 20-25% to only about 70% of the control numbers. This limited recoverability, however, persisted even after a 95 day delay period. In all cases examined maximal recoveries were achieved within 3-7 days of NGF treatment. Neuronal size analyses provided evidence for an axotomy-induced atrophy. NGF treatments, started with 1 or 2 week delays, not only reversed fully the average somal size loss but also induced an actual hypertrophy of several of those neurons. These results provide additional evidence that at least half of the apparent loss of cholinergic medial septum neurons upon axotomy is due to a loss of markers such as the transmitter-related enzyme ChAT and NGFR rather than to actual neuronal cell death. These results also show that NGF exerts a genuine trophic influence by regulating the size of its target neurons as well as their content of several proteins. PMID- 2558782 TI - Antipyretic effect of central alpha-MSH summates with that of acetaminophen or ibuprofen. AB - The neuropeptide alpha-melanocyte stimulating hormone has potent antipyretic properties when given centrally or systemically. Little is known about the mechanism of the antipyretic action and virtually nothing is known about interactions among the antipyretic effects of alpha-MSH and commonly used antipyretic agents. Randomized studies in which alpha-MSH, acetaminophen or ibuprofen, or a combination of alpha-MSH and antipyretic drug, were given IV to rabbits made febrile by endogenous pyrogen, indicate that the antipyretic effect of the peptide is summative with those of the drugs. alpha-MSH has been shown to have a wide safety margin in earlier research and the results suggest that combinations of peptide and antipyretic drugs might be safer and have fewer side effects than larger quantities of the drugs alone. PMID- 2558783 TI - Alpha-adrenergic receptor modulation of the intrathoracic efferent sympathetic nervous system regulating the canine heart. AB - The augmentation of ventricular inotropism induced by electrical stimulation of acutely decentralized efferent sympathetic preganglionic axons was reduced, but still present, following administration of hexamethonium (10 mg/kg i.v.). While hexamethonium continued to be administered, the cardiac augmentations so induced were enhanced significantly following administration of the alpha-adrenergic receptor blocking agent, phentolamine myselate (1 mg/kg i.v.). Stimulation of the sympathetic efferent postganglionic axons in cardiopulmonary nerves induced cardiac augmentations that were unchanged following administration of these agents singly or together. The cardiac augmentations induced by stimulation of efferent preganglionic sympathetic axons were unchanged when phentolamine was administered alone. The augmentations of cardiac inotropism induced by efferent postganglionic sympathetic axonal stimulation were decreased following local administration of the beta-adrenergic antagonist timolol into the ipsilateral stellate and middle cervical ganglia. Thereafter, these augmentations were unchanged following the subsequent intravenous administration of phentolamine. It is concluded that the activation of cardiac neurons in the stellate and middle cervical ganglia by stimulation of efferent preganglionic sympathetic axons can be modified by alpha-adrenergic receptors and that these effects are dependent upon beta-adrenergic receptors, not nicotinic ones, in intrathoracic ganglia. PMID- 2558784 TI - Kappa-opioid agonist induced diuresis. Is it mediated by a blood-borne factor of adrenomedullary origin? AB - Intravenous administration of the kappa-opioid agonists U50488H, tifluadom, and ethylketocyclazocine induced a characteristic diuresis in conscious, intact, saline-loaded rats. Naloxone pretreatment antagonized U50488H-induced diuresis. The diuretic response to the kappa-opioid agonists was significantly attenuated in adrenal demedullated rats. However, basal urine output, the diuretic response to furosemide, and the antidiuretic response to the mu-opioid agonist buprenorphine were unaffected. Transfusion studies established that 1 mL of blood, from intact donor rats treated with U50488H, induced a diuretic response when administered to intact or demedullated recipient rats, whether or not the recipients had been pretreated with naloxone. However, blood from demedullated rats treated with U50488H was unable to induce diuresis in intact or demedullated recipients. The results indicate that kappa-opioid agonist induced diuresis appears to be mediated by a nonopioid blood-borne "diuretic factor" of adrenomedullary origin and that this factor might be responsible for the dependence of the diuretic response upon an intact and functional adrenal medulla in conscious rats. PMID- 2558785 TI - Effect of dietary fibers on glycemia and insulinemia and on gastrointestinal function in rats. AB - The effects of purified and semipurified dietary fiber supplements on glycemia and insulinemia were measured simultaneously with their effects on digestive tract function in the rat. An insoluble fiber (cellulose) and four soluble fibers (guar gum, carboxymethylcellulose, mustard mucilage, and oat beta-glucan) were added separately to a fiber-free solid diet and fed to Sprague-Dawley rats for 10 days. Guar gum and oat beta-glucan reduced the food intake, whereas cellulose increased it. Guar gum reduced weight gain. Cellulose increased the protein efficiency ratio. After a 13-h fast, glycemia and insulinemia were measured 45, 90, 210, and 360 min after the beginning of a voluntary short meal. Addition of fibers did not change the glycemic response, but soluble fibers significantly decreased insulinemia 45 min after the meal. All fibers significantly delayed gastric emptying, cellulose and mustard mucilage being the most effective. Dry matter contents of the small intestine were increased especially by guar gum and oat beta-glucan. All fibers seemed to slow down small intestinal transit and decreased intestinal absorption. In the present experimental situation, both gastric and intestinal components played a role in the hypoinsulinic effect of dietary fibers. The intestinal component appeared to be more determinant for all soluble fibers, except mustard mucilage where the gastric component was more important. PMID- 2558786 TI - Developmental changes in the effects of carbachol and morphine on cGMP contents of plasma, heart, and lung of mice. AB - It is considered that carbachol increases plasma cGMP levels by acting on muscarinic receptors and morphine increases these levels by acting on opioid receptors, followed by stimulation of muscarinic receptors. We investigated the ability of carbachol and morphine to increase cGMP contents of plasma, heart, and lung and the guanylate cyclase activity of heart and lung homogenate in 1-, 2-, 3 , and 7-week-old mice. The increase in plasma cGMP levels induced by carbachol showed a peak at 2 and 3 weeks of age. The basal cGMP contents in heart and lung and their rise induced by carbachol, as well as the guanylate cyclase activity of these organs, were decreased in 7-week-old mice. The effects of morphine on the cGMP contents showed a similar developmental change, except for no effect in 1 week-old mice. These changes in the effects of carbachol and morphine may be the result of developmental changes of the muscarinic receptor--guanylate cyclase system and opioid receptors. PMID- 2558787 TI - Ionic mechanisms of action of GABA on dorsal and ventral root myelinated fibers: effects of K+ channel blockers. AB - Excitability changes evoked by the inhibitory neurotransmitter, GABA (gamma aminobutyric acid) in myelinated axons of dorsal and ventral roots of the isolated bullfrog sciatic nerve were compared in the absence and presence of K+ channel blockers. Half-maximal A-fiber responses to a 0.5-Hz stimulation of the whole nerve were recorded from individual roots. Direct applications of Ringer with raised K+ levels to the site of stimulation caused increases in excitability of both dorsal and ventral root fibers, which resembled those evoked in the ventral root by the GABA agonist THIP (4,5,6,7-tetrahydroisoxazolo[5,4-c]ol). The increases in dorsal root fiber responses produced by GABA were depressed by tetraethylammonium (TEA) (3 mM), 4-aminopyridine (4-AP) (50 microM), Cs (2 mM), and Ba (1 mM). Ventral root fibers were less consistently affected. The early component of GABA-evoked excitability increases was depressed by 4-AP, Cs, and Ba, but greatly augmented by TEA. THIP-evoked changes in the excitability of the dorsal and ventral root fibers were, respectively, depressed and enhanced by TEA. The augmenting effect of TEA on the early component of GABA agonist effects on the ventral root fibers is attributed to their high resting K+ conductance and the presence of a slowly inactivating, fast K+ current (If1). The depressant effects of K+ channel blockade on depolarizing components of agonist-evoked changes in dorsal and ventral root responses indicate interference with release and (or) sensitivity to K+ and a possible contribution from a mechanism involving voltage-dependent delayed rectifier K+ currents. PMID- 2558788 TI - Pharmacological characterization of the binding of [3H]bremazocine in guinea-pig brain: evidence for multiplicity of the kappa-opioid receptors. AB - In guinea-pig brain, [3H]bremazocine has a binding capacity of 27.2 pmol/g wet tissue, which is statistically different from that of [3H]ethylketazocine (14.7 pmol/g wet tissue) or the sum of the individual binding capacities of mu-, delta , and kappa-selective ligands (15.0 pmol/g wet tissue). Saturation studies of [3H]bremazocine performed in the presence of unlabelled mu-, delta-, and kappa blockers still reveal a homogeneous population of binding sites. [3H]Bremazocine under suppressed conditions displays at these sites a Kd of 2.51 nM with a binding capacity of 9.15 pmol/g wet tissue. We have performed the pharmacological characterization of these additional opioid binding sites. Displacement curves measured with a number of opioid substances were all best fitted to a one-site model. The stereoselectivity of these additional sites was demonstrated by using two groups of stereoisomers. Oripavine and benzomorphan opioids were among the most potent drugs at the [3H]bremazocine sites (mu + delta + kappa suppressed). Diprenorphine, bremazocine, cyclazocine, and ethylketazocine displayed apparent affinities constants (1/Ka) of 8.66, 7.57, 21.4, and 38.0 nM, respectively at those sites. The kappa-selective drugs U50488, U69593, PD117302, and tifluadom were inhibitors of the binding of [3H]bremazocine at these sites with apparent affinities of 113, 268, 76.9, and 47.9 nM. All mu- or delta-selective drugs tested in this study have caused weak or no inhibition of the binding. Correlation analyses were done between the different affinities measured at the [3H]bremazocine sites (mu + delta + kappa suppressed) and those observed at the known mu-, delta-, and kappa-sites of the guinea-pig brain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558789 TI - Isolation of marine bacteria with antiviral properties. AB - We report in this study the isolation of marine bacteria with antiviral properties that have been tentatively classified as Moraxella. These bacteria retained their virucidal capacity after prolonged subcultivation in the laboratory. The virus-inactivating agent could not be separated from the viable marine bacteria, indicating that the active agent(s) either remains associated to the microorganisms or has a very short lifetime, or both. The antiviral capacity of the isolated microorganisms was highly specific for poliovirus. No virucidal effect was observed against other strains of enteroviruses, such as Coxsackie and ECHO virus, rotavirus SA11, or bacteriophages proposed as indicators of the virological quality of water, such as coliphage f2 and bacteriophage B40-8, which infects Bacteroides fragilis. PMID- 2558791 TI - Decreased cytosolic free calcium concentration of aged human lymphocytes in resting state. AB - Cytosolic free calcium concentration was measured in lymphocytes from individuals over the age of 80, using quin2 and fura-2 calcium indicators. The average intracellular free calcium concentration of the samples was 62 nM, which value is roughly half the adult (age between 35 and 55) level (116 nM). It is supposed that the decline in immune function of aged individuals is connected to the decrease in free calcium concentration in their lymphocytes. We also discuss the consequences and the adaptive character of this decrease. PMID- 2558790 TI - Drugs acting on sigma and phencyclidine receptors: a review of their nature, function, and possible therapeutic importance. PMID- 2558792 TI - Renal and adrenal resistance against atrial natriuretic peptide in congestive heart failure: effect of angiotensin I-converting-enzyme inhibition. AB - We compared the natriuretic and diuretic effect of an intravenous infusion of 1 28 human atrial natriuretic peptide (hANP) (0.1 micrograms/kg/min over 30 min) in 10 patients with congestive heart failure (CHF) and in 10 control subjects of similar age and sex. In the controls, urine volume rose from 36.8 +/- 8.55 to 115.6 +/- 34.2 ml/30 min and urinary sodium excretion from 4.55 +/- 0.8 to 11.2 +/- 2.24 mEq/30 min before and during the infusion of ANP, respectively. In patients, baseline urine volume and sodium output were similar, however, rise in urine volume and urinary sodium was greatly reduced during the infusion of hANP. In patients with CHF, baseline plasma ANP levels (604.1 +/- 135.3 vs. 39.4 +/- 5.85 pg/ml; p less than 0.005) and urinary excretion of cyclic GMP (cGMP) (41.8 +/- 5.22 vs. 15.2 +/- 4.19 nmol/30 min; p less than 0.05) were significantly elevated compared to controls. The absolute and relative rise in cGMP excretion, however, was blunted in patients with CHF. In the controls, angiotensin I converting-enzyme (ACE) inhibition by enalapril significantly reduced the urinary output of sodium and water after ANP infusion. Plasma ANP levels and urinary cGMP remained unaltered by ACE inhibition. Furthermore, treatment with enalapril resulted in a rise in renin and a drop in aldosterone levels. The reduction of plasma renin and serum aldosterone by ANP was maintained after ACE inhibition. In the patient group, administration of enalapril (3 X 2.5 mg every 6 h) reduced ACE activity in the serum from 84.7 +/- 16.9 to 2.13 +/- 0.88 U/l. Arterial blood pressure was lowered from 114.7 +/- 6.69 to 106.1 +/- 7.25 mm Hg systolic and from 76.9 +/- 3 to 69.2 +/- 3.7 mm Hg diastolic. However, natriuresis and diuresis and creatinine clearance following infusion of ANP remained unaltered. PMID- 2558793 TI - Biochemical characterization of a digitalis-like substance in human urine, partially purified by high pressure liquid chromatography. AB - We have partially purified a digitalis-like extract from human urine by means of high pressure liquid chromatography (HPLC). Four fractions, giving positive activity for the cross-reaction in the digoxin radioimmunoassay, Na-K-ATPase inhibition or competition with ouabain binding to its receptor, were detected on HPLC. Among these fractions, the first (F-1) was shown to contain the principle fraction, sharing all the activities, and to exhibit quite similar characteristics to ouabain in the modes of Na-K-ATPase inhibition, ouabain displacement activity and positive inotropic action. PMID- 2558794 TI - Calcitonin gene related peptide (CGRP) activates adenylate cyclase of bovine aortic endothelial cells: guanosine 5' triphosphate dependence and partial agonist activity of a tyrosinated analogue. AB - Human calcitonin gene related peptide (hCGRP) has been shown to release prostacyclin from cultured human endothelial cells and activate adenylate cyclase in endothelial cell membrane preparations. Human endothelial cells are difficult to obtain, so a bovine aortic endothelial cell model has been used to investigate the actions of hCGRP further. Bovine aortic endothelial cells were cultured and membranes prepared from them. In these membrane fractions there was a concentration dependent activation of adenylate cyclase by hCGRP, which required the presence of guanosine 5' triphosphate. Basal activity of adenylate cyclase was 41.5 pmol.min-1.mg-1 protein. The concentration for half maximum activation of adenylate cyclase (Kact) by hCGRP was 842 nM. The maximum increase in adenylate cyclase activity above basal in response to hCGRP was 180 pmol cyclic adenosine monophosphate.min-1.mg-1 protein. A [tyr degree] substituted analogue of hCGRP [( tyr degree]-hCGRP) also activated adenylate cyclase (Kact 2.2 microM) but the maximum stimulation by [tyr degree]-hCGRP was less than that obtained with native peptide. The same analogue partially inhibited the hCGRP dependent activation of adenylate cyclase (Ki = 2 microM). These studies show that hCGRP acts on a receptor on endothelial cells to activate adenylate cyclase and that [tyr degree]-hCGPO is a partial agonist at this receptor. The potent vasodilator properties of hCGRP may be mediated through or modified by its interaction with endothelial cells. PMID- 2558795 TI - Electrophysiological effects of FK664, a new cardiotonic agent, on preparations from guinea pig ventricle and from rabbit sino-atrial node. AB - Effects of the cardiotonic agent FK664, 6-(3, 4-dimethoxy-phenyl)-1-ethyl-4 mesitylimino-3-methyl-3,4-dihydro-2 (1H)-pyrimidone, on isolated guinea pig ventricular muscles and rabbit sinus node pacemaker cells were studied using micro-electrode techniques. In ventricular muscles driven at 0.5-1.0 Hz, FK664 above 3 mumol.litre-1 caused an increase in contractile force and a shortening of time to peak tension. This positive inotropic effect of FK664 was accompanied by a slight elevation of the early plateau phase of the action potential, while other action potential variables were unaffected. The change in contractile force induced by FK664 was abolished in a low Ca2+ medium (0.12 mmol.litre-1) or by treatment with ryanodine (2 mumol.litre-1), whereas it was relatively well preserved in the preparations pretreated with nefedipine (1 mumol.litre-1). The slow action potentials induced by isoprenaline (0.3 mumol.litre-1) in high K+ medium (30 mmol.litre-1) and the slow inward current measured by single sucrose gap voltage clamp at a holding potential of -40 mV were unaffected by FK664. In sinus node pacemaker cells, FK664 (1-10 mumol.litre-1) caused a dose dependent acceleration of phase 4 depolarisation and a shortening of spontaneous firing cycle length. This positive chronotropic effect of FK664 was markedly inhibited in a low Ca2+ medium (0.3 mmol.litre-1). These findings suggest that FK664 has positive inotropic and chronotropic effects on the heart, due to an enhancement of transsarcolemmal calcium influx through the low threshold, dihydropyridine insensitive Ca2+ channel population. PMID- 2558796 TI - An increase in myocardial beta-adrenoceptors to compensate for postischaemic dysfunction following coronary micro-embolisation in dogs. AB - This study examined whether beta-adrenoceptors increase in number during recovery from prolonged myocardial stunning and whether they compensate for lack of physiological response to beta-adrenergic stimulation in this abnormality. The left coronary artery was embolished in anaesthetised dogs with non-labelled microspheres (15 +/- 1 micron; 1.2 X 10(6).kg-1 body weight). Haemodynamic studies were performed before (control) and 24 h and 1 week after embolisation, in the conscious state. Myocardial noradrenaline content, plasma catecholamine concentrations and the density of beta-adrenoceptors (Bmax) were also assessed at three study intervals. At 24 h after embolisation, both systolic and diastolic cardiac function was significantly depressed. The inotropic response to isoprenaline was preserved, but the response to forskolin was markedly depressed. One week after embolisation, resting systolic function was restored to control levels and histological examination showed absence of myocardial necrosis. Although plasma noradrenaline concentration had returned to normal, myocardial noradrenaline content had decreased by 36% and the density of beta-adrenoceptors had increased by 48%. Myocardial relaxation was still impaired and the inotropic response to forskolin was also still depressed, whereas the response to isoprenaline was normal. Moreover, the down regulation of the increased beta adrenoceptors by isoprenaline infusion for 24 h unmasked the latent systolic dysfunction. These results indicate that the density of beta-adrenoceptors increases during the recovery process from prolonged myocardial stunning and that this increase may compensate, at least in part, for impairment of the inotropic mechanism distal to the beta-adrenoceptors. PMID- 2558797 TI - Effects of enalapril on the cardiovascular response to treadmill exercise in patients with mild to moderate systemic hypertension. AB - The effects of the angiotensin converting enzyme inhibitor enalapril on exercise induced changes in blood pressure and heart rate were evaluated in 15 patients in the early stages of systemic hypertension. Multistage treadmill exercise was performed before and after eight weeks of enalapril administration, and the results of the two trials were compared. In patients at rest, enalapril decreased systolic blood pressure from 172 +/- 18 to 147 +/- 14 mmHg and diastolic blood pressure from 99 +/- 9 to 88 +/- 8 mmHg (both P less than 0.001). In patients at peak exercise, enalapril decreased systolic blood pressure from 216 +/- 13 to 195 +/- 18 mmHg and diastolic blood pressure from 106 +/- 12 to 99 +/- 12 mmHg (both P less than 0.001). There was also a significant decrease in blood pressure during the recovery period after treadmill exercise. Enalapril reduced heart rate at peak exercise (P less than 0.05), but not at rest or during recovery. Thus enalapril alleviated the response of blood pressure to exercise in hypertensive patients and may help prevent hypertensive complications during daily activities. PMID- 2558799 TI - Repository corticotropin injection as an adjunct to smoking cessation during the initial nicotine withdrawal period: results from a family practice clinic. AB - Fifteen white patients participated in this study of a family-practice-based smoking cessation program in which corticotropin (ACTH) was used to assist patients during the first one to two weeks of abstinence from nicotine. All patients were habitual smokers who had made one or more attempts to quit smoking before entering this program. Treatment consisted of three single intramuscular injections of ACTH. In most cases, declining doses of 160, 80, and 40 U were administered at three-day intervals. The decision to provide additional treatment was based on the response by each volunteer and the investigator's judgment. The mean duration of follow-up was 33 days (range, 17 to 49 days), with the exception of two patients, who have just recently completed therapy. Evidence supporting complete smoking cessation or a significant reduction (80% to greater than 90%) in the number of cigarettes smoked per day was achieved in 13 of the 15 patients. The single nonresponder expressed an initial interest in the program, but showed a lack of motivation thereafter. There was insufficient follow-up on one other patient, who has not been available for monitoring since completion of therapy. Symptoms associated with the tobacco withdrawal syndrome that were reported by the patients included mild irritability and restlessness. PMID- 2558800 TI - Discrepancy in hormone binding and information transfer between sister cells of Tetrahymena clones. AB - Tetrahymena cells treated with insulin in mass cultures were separated to single cell clones or one of the "sister-cells" of dividing Tetrahymena (in single-cell culture) was treated with insulin. In both cases the FITC-insulin binding of sister-cells were compared. The insulin imprinting significantly increased the insulin binding of cells. There was also a significant difference between the imprinted and not imprinted sisters as well as between the not imprinted sisters. This demonstrates the existence of a difference (in hormone binding) between sister-cells and justifies that the information of the first hormone treatment (imprinting) is not equally divided between the sister-cells. PMID- 2558798 TI - Nicotine, corticotropin, and smoking withdrawal symptoms: literature review and implications for successful control of nicotine addiction. AB - The literature on corticotropin (ACTH) regulation of cortisol secretion, effects of nicotine on plasma cortisol and ACTH levels, and the cortisol response to smoking cessation is reviewed. A dose-dependent increase in plasma cortisol levels is found in habitual smokers after smoking two cigarettes. Nicotine mediates this response via a central mechanism to release cortisol from the adrenal cortex. The sites of action of nicotine appear to be in the hypothalamus or brain stem. Complete cessation of smoking is followed by a fall in plasma cortisol levels that is associated with the withdrawal of the nicotine stimulus. In addition, anxiety levels increase and plasma epinephrine levels decrease. Nicotine withdrawal symptoms confront smokers who want to quit. Many of the symptoms seem to be related to the body's response to changes in cortisol levels. As part of a comprehensive smoking cessation program, one or two intramuscular injections of ACTH gel have been shown to help smokers stop and continue to abstain from smoking. PMID- 2558801 TI - Cytochemical demonstration of glucose-6-phosphatase in human liver. AB - To determine the cytochemical localization of glucose-6-phosphatase in the human hepatocyte, lead - based and cerium - based media were used. By studying the effects of systematic variation of the incubation medium components, the optimal experimental conditions were determined. The exclusive localization of the cytochemical reaction in the endoplasmic reticulum and nuclear envelope, together with the results of control experiments ensured that these findings could be correlated with the phosphohydrolase activity of the multicomponent glucose-6 phosphatase system. PMID- 2558803 TI - Receptor autoradiographic analysis of insulin-like growth factor-I (IGF-I) binding sites in rat forebrain and pituitary gland. AB - 1. Specific 125I-labeled insulin-like growth factor-I [( 125I] IGF-I) binding sites in the rat forebrain and pituitary gland were investigated using quantitative receptor autoradiography. 2. High densities of [125I]IGF-I binding sites were present in the olfactory nerve layer, olfactory glomerular layer, choroid plexus, CA3 and CA4 of the hippocampus, basolateral amygdaloid nucleus, and endopiriform nucleus. Moderate to high binding densities were found in the cerebral cortex (II, VI), bed nucleus stria terminalis, accumbens nucleus, lateral septum, median preoptic nucleus, supraoptic nucleus, paraventricular hypothalamic nucleus, and ventroposterior thalamic nucleus. In the circumventricular organs, subfornical organ, vascular organ of the lamina terminalis, and median eminence, the binding sites were numerous. High densities of [125I]IGF-I binding sites were also observed in the anterior pituitary gland. 3. In kinetic experiments, [125I]IGF-I binding sites in the olfactory glomerular layer, choroid plexus, median eminence, and anterior pituitary gland were found to be single and of a high affinity. 4. Noteworthy was the difference in the potency of insulin in inhibiting the binding among the areas examined, a finding which suggests heterogeneity of IGF-I receptors. 5. The possibility that IGF-I plays the role of a neurotransmitter and/or neuromodulator in the central nervous system warrants further investigation. PMID- 2558802 TI - Binding sites for brain trace amines. AB - 1. Neurochemical, neuropharmacological, and neurophysiological studies suggest that some of the so-called trace amines may have a role in the modulation of neurotransmission. This review examines the possible existence and characterization of brain binding sites for the trace amines. 2. The results of radioligand binding studies carried out so far suggest the existence of tryptamine binding sites that possibly constitute a true functional receptor. This is supported by evidence obtained from the saturation studies, drug-mediated inhibition of binding, and the changes in the number of sites induced by pharmacological and lesion studies. In addition, the existence of a functional tryptamine binding site is supported by the increased neurophysiological responses of tryptamine obtained from the striatum of rat with unilateral substantia nigra lesions. 3. It has been shown that the brain contains saturable binding sites for rho-tyramine that appear to be related to the transport of dopamine into synaptic vesicles. There are, however, some questions with respect to the homogenization technique employed and some inconsistencies with respect to the number of binding sites estimated in neuronal membrane preparations. 4. The existence of rho-octopamine binding sites has been demonstrated in crude membranes obtained from fruitflies but not shown so far in vertebrates. 5. The presence of brain binding sites for beta-phenylethylamine are suggested but they are not so well defined and its physiological implication remains to be elucidated. PMID- 2558805 TI - An autoradiographic study of alpha 1-adrenergic receptors in the brain of the Japanese quail (Coturnix coturnix japonica). AB - The neuroanatomical distribution of alpha 1-adrenergic receptors was studied in Japanese quail by quantitative in vitro autoradiography using the specific antagonist [3H]prazosin as the ligand. The presence of saturable (Bmax less than 200 fmol/mg protein) high affinity (Kd less than 0.12 nM) binding sites was detected by saturation analysis. High concentrations of [3H]prazosin binding sites were detected in the archistriatum/pars ventralis, the hippocampus, the cortex piriformis, the area corticoidea dorsolateralis, the dorsal thalamus, and the nucleus praetectalis. Lower concentrations were seen in the intercollicular nucleus, the lateral septum, and the posterior and tuberal hypothalamus. Very little binding was seen in the preoptic and anterior hypothalamic areas. The relatively high number of binding sites identified in the telencephalic structures agrees well with previous mammalian studies. This is in contrast with the pattern in the anterior hypothalamus where, in mammals, a number of nuclei have been reported to contain a high receptor density. PMID- 2558807 TI - [A serological study on anti-CMV and anti-EBV antibodies in population of Beijing, Changzhi Shanxi and Yichang Hubei]. AB - 1358 serum samples of healthy persons in Beijing (785) Changzhi, Shanxi (309) and Yichang, Hubei (264) of different age groups were detected for anti Cytomegalovirus (CMV) IgG and anti-Epstein-Barr virus (EBV) IgG antibodies by using ELISA. The positive rates and GMT of anti-CMVIgG antibody in each group were 88.8%, GMT 1:1661.6; 89.4%, GMT 1:749.9; 98.4%, GMT1:1448.2 respectively. The positive rates and GMT of anti-EBVIgG antibody in each group were 98.1%, GMT 1:258.8; 94.3%, GMT 1:99.2; 100%, GMT 1:539.4 respectively. The data showed that there were significant differences in the positive rates between the urban and rural areas. PMID- 2558806 TI - Ca2+/calmodulin-dependent protein phosphatase in bovine parotid gland: purification and characterization. AB - Calmodulin-dependent protein phosphatase (CaM-PPase) was isolated from bovine parotid gland by sequential application of DEAE-52, Affi-gel blue and calmodulin affinity chromatography followed by gel filtration and high performance liquid chromatography. The enzyme was activated in the simultaneous presence of Ni2+ or Mn2+ and Ca2+ plus calmodulin. Ca2+/calmodulin-dependent activation of CaM-PPase was antagonized by inhibitors of calmodulin action, such as W-7 and trifluoperazine. Tryptophan fluorescence was quenched in the presence of Ni2+. CaM-PPase was a heterodimer. The molecular weights of large subunits which bound calmodulin (CaM) were 68 kD and 58 kD - the 68 kD subunit was predominant. Polyclonal antibodies against bovine calcineurin cross-reacted with both types of larger subunits. Using polyclonal antibodies against bovine calcineurin or the monoclonal antibody against subunit B of bovine calcineurin, the smaller molecular weight subunit (19 kD) was found to be immunologically identical to subunit B of bovine calcineurin. In bovine parotid gland, CaM-PPase was found both in acinar and duct cells. PMID- 2558804 TI - Enzymatic inactivation of bradykinin by rat brain neuronal perikarya. AB - 1. Bradykinin (Bk; Arg1-Pro2-Pro3-Gly4-Phe5-Ser6-Pro7-Phe8-Arg8) inactivation by bulk isolated neurons from rat brain is described. 2. Bk is rapidly inactivated by neuronal perikarya (4.2 +/- 0.6 fmol/min/cell body). 3. Sites of inactivating cleavages, determined by a kininase bioassay combined with a time-course Bk product analysis, were the Phe5-Ser6, Pro7-Phe8, Gly4-Phe5, and Pro3-Gly4 peptide bonds. The cleavage of the Phe5-Ser6 bond inactivated Bk at least five fold faster than the other observed cleavages. 4. Inactivating peptidases were identified by the effect of inhibitors on Bk-product formation. The Phe5-Ser6 bond cleavage is attributed mainly to a calcium-activated thiol-endopeptidase, a predominantly soluble enzyme which did not behave as a metalloenzyme upon dialysis and was strongly inhibited by N-[1(R,S)-carboxy-2-phenylethyl]-Ala-Ala Phe-p-aminobenzoate and endo-oligopeptidase A antiserum. Thus, neuronal perikarya thiol-endopeptidase seems to differ from endo-oligopeptidase A and endopeptidase 24.15. 5. Endopeptidase 24.11 cleaves Bk at the Gly4-Phe5 and, to a larger extent, at the Pro7-Phe8 bond. The latter bond is also cleaved by angiotensin converting enzyme (ACE) and prolyl endopeptidase (PE). PE also hydrolyzes Bk at the Pro3-Gly4 bond. 6. Secondary processing of Bk inactivation products occurs by (1) a rapid cleavage of Ser6-Pro7-Phe8-Arg8 at the Pro7-Phe8 bond by endopeptidase 24.11, 3820ACE, and PE; (2) a bestatin-sensitive breakdown of Phe8 Arg9; and (3) conversion of Arg1-Pro7 to Arg1-Phe5, of Gly4-Arg9 to both Gly4 Pro7 and Ser6-Arg9, and of Phe5-Arg9 to Ser6-Arg9, Phe8-Arg9, and Ser6-Pro7, by unidentified peptidases. 7. A model for the enzymatic inactivation of bradykinin by rat brain neuronal perikarya is proposed. PMID- 2558808 TI - Two genes encode 7SL RNAs in the yeast Yarrowia lipolytica. AB - We have identified an abundant cytoplasmic 7S RNA in crude extracts of the yeast Yarrowia lipolytica. A cDNA probe was prepared from this RNA and used to screen a genomic library. The DNA sequence of a positive clone was determined and the end positions of the 7S RNA gene established by comparison with the sequence of the extremities of 7S RNA. This gene, designated SCR2, encodes a 270-nucleotide RNA that can be folded into a secondary structure similar to that of 7SL RNAs. This RNA is 94.4% homologous to a previously identified 7S RNA from this yeast, but is encoded by a separate gene with highly divergent flanking sequences. PMID- 2558810 TI - High-performance liquid chromatography of arachidonic acid metabolites and its application to the determination of leukotriene B4 in stimulated leukocytes. AB - A highly sensitive high-performance liquid chromatography with fluorescence detection for the determination of arachidonic acid metabolites is described. The metabolites are converted into corresponding fluorescent derivatives by reaction with 3-bromomethyl-6,7-methylenedioxy-1-methyl-2(1H)-quinoxalinone in the presence of potassium hydrogen carbonate and 18-crown-6 in acetonitrile. The derivatives are separated on a reversed-phase column (Inertsil ODS) with aqueous acetonitrile and detected fluorimetrically. The detection limits are 5-15 fmol at a signal-to-noise ratio of 3 in a 10-microliter injection volume. The method is applied to the determination of leukotriene B4 produced in stimulated leukocytes. PMID- 2558811 TI - [In vivo study of the inhibitory effect of sodium selenite on mouse complement activation]. AB - A simple and sensitive rocket immunoelectrophoresis technique for the assessment of complement activation was employed to study the effect of sodium selenite on mouse complement activation. It was shown that selenite can cause both in vitro and in vivo inhibition of mouse complement activation. The most significant effect was observed in the inulin-activated group; and not in the mouse aggregated IgG activated group. These results suggest that selenite has some inhibiting effect on the alternative complement activation pathway but not on the classical pathway. Therefore, sodium selenite may be applicable in the alleviation or prevention of symptoms of diseases accompanied by alternative pathway complement activation. PMID- 2558812 TI - [The in vitro effect of silica fibers on macrophage-mediated cytotoxic activities in mice]. AB - The in vitro effects of silica fibers on macrophage-mediated cytotoxic activities in mice were studied, including 1) macrophage-mediated tumor cytolysis (MTC), 2) macrophage-mediated tumor cytostasis (MTCS) and 3) antibody-dependent cell mediated cytolysis (ADCC), we found that silica fibers markedly inhibited both MTC and ADCC but slightly promoted MTCS. Silica fibers were active during the activation phase mediated by macrophage-activating factor as well as in the effector phases of MTC and MTCS. In ADCC, increasing the antibody titer could not abrogate the inhibitory effect of silica fibers. PMID- 2558809 TI - DNA sequence analysis of the 24.5 kilobase pair cytochrome oxidase subunit I mitochondrial gene from Podospora anserina: a gene with sixteen introns. AB - The DNA sequence of a 26.7 Kilobase pair (10(3) base pairs = 1 Kb) region of the mitochondrial genomes of races s and A from Podospora anserina was determined. Within this region, the 24.5 Kb cytochrome oxidase subunit I gene was located and its exon sequences determined by computer analysis comparisons with other fungal genes. The Podospora COI gene was interrupted by two group II introns (one in race s) and fourteen group I introns ranging in size from about 2.2 Kb to 404 bp. Earlier studies on secondary structure analysis, as well as comparison of their open reading frames (ORFs), showed that the two group II introns were closely related. The fourteen group I introns were representatives of three subgroupings (IB, C and a new category, subgroup ID). Two of these group I introns were separated by just a single exon codon. The analysis of all these introns is discussed in comparison with other fungal introns as well as with the known Podospora anserina introns. PMID- 2558813 TI - Characterization of dolichyl diphosphate phosphatase from rat liver. AB - Dolichyl diphosphate phosphatase (DolPPase) has been characterized in rat liver. Subcellular distribution studies indicate that the enzyme is localized in the endoplasmic reticulum. The in vitro enzymatic activity is stimulated by EDTA, due to release of inhibition by trivalent cations found in the assay tubes. All di- and trivalent cations tested were inhibitory, with the trivalent ions Al3+ and Fe3+ showing greater than 70% inhibition at a concentration of 10 microM. The assay requires the presence of a detergent for optimal activity, with Triton X 100 giving maximum activity at 0.1%. The substrate specificity of DolPPase toward polyprenyl diphosphates has been determined and indicates that there is little preference of the enzyme for substrates of different chain length, and either stereochemical orientation or degree of saturation of the alpha-isoprene unit. Km values of 11-14 microM were obtained for all substrates tested. Preliminary studies on the transmembrane topology of the DolPPase using latency assays, indicate that the active site of the enzyme may reside on the cytoplasmic face of the endoplasmic reticulum. PMID- 2558815 TI - Adrenocortical activity and plasma cAMP levels of humans under high altitude stress. PMID- 2558814 TI - Absence of "A"-esterase activity in the serum of a patient with Tangier disease. AB - The levels of apolipoprotein A-I, A-II and B in subjects who are homozygous or heterozygous for Tangier disease are reported and compared with the amount of "A" esterase in the serum. The "A"-esterases hydrolyse toxic organophosphate pesticides and are currently classified by the nomenclature committee of the International Union of Biochemistry as arylesterases (EC 3.1.1.2) although recent evidence has cast doubt on this classification. The apolipoprotein data are consistent with previous data reported for a number of Tangier patients. The homozygote has a marked reduction in apo A-I and A-II levels and a 30% reduction in apo B. The heterozygotes have about a 50% reduction of apo A-I, a slight reduction in apo A-II and no change in apo B. These apolipoprotein values correspond to a marked reduction in HDL cholesterol for the homozygote and substantial reductions in the heterozygotes. The "A"-esterase activity is zero in one homozygote while heterozygotes have about 5% of the levels in control subjects. Arylesterase activity appears to be essentially normal. The data thus support previous observations that the HDL "A"-esterase activity is greatly reduced in those conditions where HDL apo A-I is markedly reduced, e.g., in "Fish eye" Disease. PMID- 2558816 TI - Bilateral inferior petrosal sinus sampling as a routine procedure in ACTH dependent Cushing's syndrome. AB - Bilateral inferior petrosal sinus sampling was successfully performed in 12 of 13 consecutive patients with ACTH-dependent Cushing's syndrome. Ten of the patients subsequently had transsphenoidal pituitary microsurgery. Eight patients in whom the inferior petrosal sinus to peripheral vein ACTH level ratio was 1.5 or greater were found to have a pituitary adenoma. One of the remaining two patients who had ratios less than 1.5 had pituitary hyperplasia while the other had no identified abnormality. In five of the patients with pituitary tumour a ratio above 1.5 was present on only one side. Bilateral petrosal sampling is therefore always necessary. Tumour localization within the pituitary was only poorly predicted by either petrosal sinus sampling (four of eight) or computed tomography scanning (three of eight). If petrosal sinus sampling is used early in the differential diagnosis of ACTH-dependent hypercortisolism, then the use of other differential diagnostic tests may not always be necessary. PMID- 2558817 TI - The effect of ovine corticotrophin-releasing factor on the hormonal response to insulin-induced hypoglycaemia. AB - In order to investigate the role of hypothalamic corticotrophin-releasing factor (CRF41) in the mediation of the pituitary ACTH response to hypoglycaemia, eight normal adult males were studied on four occasions. Commencing at 0830 h after an overnight fast, each received, in double-blind, random order, intravenous boluses of: A, normal saline control; B, soluble insulin 0.15U/kg; C, ovine CRF41 (oCRF41) 100 micrograms; D, soluble insulin 0.15U/kg followed immediately by oCRF41 100 micrograms. Adequate hypoglycaemia (blood glucose less than 2.2 mmol/l) was achieved in each subject when insulin was given alone or with oCRF41, and there was no difference in the glucose nadir between the 2 days. Peak plasma ACTH was significantly higher after insulin plus oCRF41 than after insulin alone (P less than 0.05) or oCRF41 alone (P less than 0.01) and this enhancement of ACTH release was most marked in the first phase of the response at 30 min (P less than 0.001, b vs d). There was no difference in the peak serum cortisol response whether oCRF41 and insulin were given alone or together and although the area under the cortisol curve was greater after insulin plus oCRF41, this difference was explicable simply on the basis of the earlier onset of the cortisol response to oCRF41. There were no differences in the serum GH responses to hypoglycaemia on the 2 days.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558818 TI - Neurocirculatory regulation in cortisol-induced hypertension. AB - Effects of chronic glucocorticoid treatment on arterial baroreflex function and on cardiac beta- and vascular alpha-adrenoceptor-mediated responses were assessed in conscious, unrestrained Wistar-Kyoto rats. Cortisol (25 mg/kg/day) was administered for seven days using a subcutaneous reservoir pump. Arterial baroreflex-cardiac sensitivity was assessed by examining the relationship of the cardiac interbeat interval to the mean arterial blood pressure during phenylephrine or nitroprusside challenge; baroreflex-sympathoneural sensitivity was assessed from the ratio of the increase in the arterial norepinephrine concentration to the decrease in mean arterial pressure at 15 min during intravenous infusion of nitroprusside; cardiac beta-adrenoceptor-mediated responsiveness was estimated from heart rate responses to bolus-injected isoproterenol; and vascular alpha-adrenoceptor-mediated responsiveness was estimated from peak mean arterial pressure responses to bolus-injected phenylephrine. Cortisol treatment increased mean arterial pressure, decreased heart rate, and increased heart rate responses to isoproterenol, whereas baroreflex-vagal sensitivity, baroreflex-sympathoneural sensitivity, and pressor responses to phenylephrine were unaffected. The results indicate that hypertension due to chronic cortisol administration is not associated with decreased sensitivity of the baroreceptor-cardiac reflex. Baroreflex sympathoneural sensitivity and alpha 1-adrenoceptor responsiveness also remain normal, whereas beta-adrenoceptor responsiveness is increased. The findings suggest that the pattern of neurocirculatory adjustment in glucocorticoid hypertension differs from that seen in other forms of hypertension. PMID- 2558819 TI - Measurement of angiotensin II concentration in rat plasma: pathophysiological applications. AB - A very reliable isocratic reverse phase high performance liquid chromatography (HPLC) system has been developed to separate angiotensins, which combined with a very sensitive radioimmunoassay, provided precise measurements of the endogenous angiotensin II (AII) concentration in the rat plasma in different experimental circumstances. The overall recoveries of AII were 95.2 +/- 15.8% (means +/- SD) when 10 pg of this peptide was added to 1 ml of human plasma. The coefficient of variation for within-assay precision was 10% (n = 6). The plasma AII, measured by this method, of normal male pentobarbital-anesthetized rats was 53.0-141.6 pg/ml (mean: 103.9 +/- 29.7 pg/ml). The plasma AII of rats fed a sodium deficient diet was 300.0 +/- 100.6 pg/ml, while that of rats given oral Enalapril, an angiotensin converting enzyme (ACE) inhibitor, for 1 week was 35.7 +/- 28.0 pg/ml. The plasma AII of bilaterally nephrectomized rats was 2.7 +/- 2.9 pg/ml 24 hours after nephrectomy and below the detection limit 48 hr after nephrectomy. This method, therefore, can be used to study AII in different pathophysiological states or after treatment with renin-angiotensin system inhibitors. PMID- 2558820 TI - Juvenile nasopharyngeal angiofibroma in a static population: the implications of misdiagnosis. AB - We present a 20-year review of juvenile angiofibroma in the relatively static population of Northern Ireland. Seventeen cases were identified and new slides were prepared from the stored paraffin blocks of all their original biopsy material, and re-examined. Five females, a 36-year-old and an 18-year-old male had their diagnoses revised. We suggest clinical criteria, which in conjunction with radiological investigations, should be strictly applied in all cases. Such application would, in retrospect, have identified those cases excluded by pathological re-examination, thus avoiding unnecessary surgery and radiotherapy. Atypical cases which do not satisfy the clinical criteria may be subjected to repeat biopsy but routine initial biopsy is not recommended. PMID- 2558821 TI - Staphylococcus aureus-stimulated mononuclear leucocyte-conditioned medium increases the neutrophil bactericidal activity, and augments oxygen radical production and degranulation in response to the bacteria. AB - Conditioned medium produced by human mononuclear leucocytes (MNL) stimulated with formalin-fixed, heat-killed Staphylococcus aureus enhanced the killing of opsonized S. aureus by human neutrophils. This enhancement was not seen when conditioned medium from non-stimulated MNL or medium cultured in the absence of both bacteria and MNL was used. There was a five-fold decrease in survival of bacteria when neutrophils were treated with conditioned medium. Conditioned medium-treated neutrophils showed increased initial rate of killing but after 20 30 min the rate of killing was similar to that of non-conditioned medium-treated leucocytes. The neutrophils treated with conditioned medium showed increased production of chemiluminescence, superoxide and lysosomal enzyme release (from both azurophilic and specific granules) in response to opsonized S. aureus. The results demonstrate that bacterial interaction with MNL leads to the release of mediators (cytokines) which potentiate the anti-bacterial function of neutrophils. PMID- 2558822 TI - IgG and IgA enhance the chrysotile-induced production of reactive oxygen metabolites by human polymorphonuclear leucocytes. AB - Chrysotile asbestos fibres induce a rapid generation of reactive oxygen metabolites by human polymorphonuclear leucocytes (PMNL) in vitro. This effect was markedly enhanced by the presence of 10-200 micrograms/ml of human gammaglobulin, purified polyclonal IgG, and monoclonal IgG and IgA myeloma proteins. Purified monoclonal IgD, IgM, kappa light chain proteins, and secretory IgA inhibited this chrysotile-induced response. No enhancing effect of IgG was observed when quartz dust or opsonized zymosan were used as stimulators of PMNL metabolism. The enhancing effect of IgG was shown to depend on opsonization of the asbestos fibre. We suggest that the IgG and IgA potentiating effect on the asbestos fibre-induced production of tissue-damaging reactive oxygen metabolites by inflammatory cells is dependent on a particle-specific binding of immunoglobulin to the fibre surface, with subsequent Fc receptor-mediated effects on cells. Such an interaction between certain immunoglobulins and asbestos may explain a number of in vivo phenomena in which immunological responses (hypergammaglobulinemia, circulating immune complexes, etc.) have been shown to relate to the progression of pulmonary asbestosis. The differences between various immunoglobulin classes and monoclonal immunoglobulins could represent an individual inflammation-modulating mechanism in the development of acute or chronic pulmonary asbestosis. PMID- 2558823 TI - Cells producing antibody to measles and herpes simplex virus in cerebrospinal fluid and blood of patients with multiple sclerosis and controls. AB - The B cell response against measles and herpes simplex virus (HSV) was evaluated in cerebrospinal fluid (CSF) and peripheral blood from patients with multiple sclerosis (MS) and controls by enumeration of cells secreting anti-measles and anti-HSV antibodies of IgG, IgA and IgM isotypes. We used a nitrocellulose immunospot assay which enables parallel enumeration of numbers of cells secreting total IgG, IgA and IgM. Anti-measles IgG antibody-secreting cells were present in CSF from 21 of 24 MS patients (mean 24 cells/10(4) mononuclear cells), and against HSV in CSF from seven of eight patients (mean 23/10(4) cells). No antibody-secreting cells were detectable in the patients' blood. Ten MS patients examined were negative for cells in CSF and blood producing anti-measles antibodies of IgA and IgM isotypes. Anti-measles IgG antibody secreting cells were also found in CSF from four of 18 controls, and anti-HSV IgG antibody secreting cells in six of 13, especially in patients with subacute or chronic inflammatory nervous system diseases. Our results confirm that viral antibodies in MS are produced within CSF and that this B cell response is preferentially sequestered to this compartment. Whether this viral B cell response in MS reflects specific activation due to persistence of viral antigens or an epiphenomenon remains to be clarified. PMID- 2558824 TI - Virus infection induces redistribution and membrane localization of the nuclear antigen La (SS-B): a possible mechanism for autoimmunity. AB - To investigate the possibility that anti-La (SS-B) antibodies in Sjogren's syndrome were induced by virus infection we studied the distribution of La in virus-infected human cell lines. Three monoclonal antibodies to La were used with monoclonal anti-Sm (derived from MRL/lpr lupus mice) and anti-rat immunoglobulin antibodies as controls. In uninfected cells La was predominantly in the nucleus. Twenty-four hours after infection of HEp-2 cells with adenovirus 2, the La and Sm antigens appeared to aggregate and accumulate in the periphery of the nucleus and, after 48 h, La was seen in the cytoplasm and cell membrane. No cytoplasmic or membrane expression of Sm was seen. Infection with adenovirus or cytomegalovirus caused a 2-13-fold increase in the concentration of La in three cell lines. Treatment of HE--2 cells with interferon-gamma (IFN-gamma) and infection with Epstein-Barr virus and cytomegalovirus caused cytoplasmic, but no definite membrane expression of La. The appearance of La on the surface of virally infected epithelial cells together with IFN-gamma induced class II expression could form the basis of a T cell dependent mechanism for anti-La autoantibody induction. PMID- 2558825 TI - Arrhythmogenic effect of forskolin in the isolated perfused rat heart: influence of nifedipine or reduction of external calcium [corrected]. AB - 1. This study investigated first the effects of forskolin on cardiac rhythm, and second the roles of calcium in cardiac arrhythmogenesis by cAMP. 2. Two series of experiments were performed. In the first series, forskolin was administered into the isolated perfused rat heart. In the second series, forskolin administration was preceded by administration of nifedipine, a calcium channel blocker, or infusion of a low concentration calcium solution. In both experiments, the myocardial cAMP level and electrocardiogram were determined. 3. It was found that forskolin increased cAMP level as well as inducing arrhythmia. Pretreatment with nifedipine or a reduction of external calcium, that either maintained or further enhanced the forskolin-induced increase in the cAMP level, abolished the forskolin-induced arrhythmia. 4. The results of the present study support the hypothesis that myocardial cAMP mediates cardiac arrhythmia, and provide evidence that calcium is essential in arrhythmia mediated by cAMP. PMID- 2558826 TI - Adrenocortical steroid requirements for initiation of ACTH-dependent hypertension in sheep. AB - 1. Previous studies demonstrated that the combined infusion of cortisol (F), aldosterone (ALDO), deoxycorticosterone (DOC), corticosterone (B), 11 deoxycortisol (S), 17 alpha-hydroxyprogesterone (17 alpha OHP) and 17 alpha, 20 alpha- dihydroxy-4-pregnane-3-one (17 alpha 20 alpha OHP), at rates equivalent to their production during adrenocorticotrophic hormone (ACTH) treatment, reproduced the pressor and metabolic responses to ACTH administration in sheep. 2. This study examined which of these adrenocortical steroids were necessary for the initiation of the hypertension produced by these steroids in sheep. 3. Infusion of F, ALDO, 17 alpha OHP and 17 alpha 20 alpha OHP together, increased MAP by 19 mmHg, similar to both complete steroid cocktail (+25 mmHg) or ACTH administration (+21 mmHg). Infusion of F, 17 alpha OHP and 17 alpha 20 alpha OHP increased MAP by +7 mmHg. Infusion of ALDO, 17 alpha OHP and 17 alpha 20 alpha OHP had no effect on MAP. Thus F and ALDO were essential for the pressor effects of the steroid infusion. 4. To determine the role of glucocorticoid activity in the MAP rise, prednisolone, a non-pressor glucocorticoid, was substituted for cortisol. Combined prednisolone, ALDO, 17 alpha OHP and 17 alpha 20 alpha OHP infusion did not raise blood pressure. This suggested that the mineralocorticoid component rather than glucocorticoid component of cortisol's activity was involved in the pressor response. 5. Aldosterone (7 micrograms/h) was substituted for cortisol, giving a total of 10 micrograms/h aldosterone. High dose ALDO (10 micrograms/h), 17 alpha OHP and 17 alpha 20 alpha OHP infusion raised blood pressure by 18 mmHg. Thus, the essential role of cortisol appeared to be due to its occupancy of mineralocorticoid receptors, rather than glucocorticoid receptors. 6. Given that ACTH produces a transient initial increase in aldosterone secretion of up to 10 micrograms/h, it appears that aldosterone and not cortisol is essential for the pressor effects of ACTH. 7. Hypertension resulting from the combined steroid infusion in the sheep appears to be produced by a mechanism which involves a complex interaction between ALDO, F, 17 alpha OHP and 17 alpha 20 alpha OHP. Therefore, the putative 'hypertensinogenic' receptor may be multivalent with binding sites for F, ALDO and 17 alpha 20 alpha OHP, or is a site of single interactive receptors for these steroids and that F exerts its permissive action by occupying the same site as ALDO on the hypertensinogenic receptors. PMID- 2558827 TI - CV 6209 is a non-competitive antagonist of platelet-activating factor receptors on guinea-pig resident peritoneal macrophages. AB - 1. The characteristics of antagonism of platelet-activating factor (Paf) receptors by the phospholipid Paf analogue, CV 6209, were studied in rabbit platelets and polymorphonuclear leucocytes (PMN) and in guinea-pig macrophages. 2. Paf-induced aggregation of PMN or platelets was antagonized in a competitive and specific manner by CV 6209 with no detectable difference between the pA2 values (approximately 9.5). 3. The specificity of CV 6209 (1-100 nmol/L) for Paf receptors in platelets and PMN was indicated by a lack of effect on A23187 (10 mumols/L) or fMLP (1 mumol/L) induced aggregation, respectively. 4. CV 6209 (1 100 nmol/L) was also a potent antagonist of Paf-induced prostacyclin (PGI2) generation by guinea-pig peritoneal macrophages. However, CV 6209 caused significant depression of the maximum response to Paf and a non-parallel shift in the concentration-response curve indicating a non-competitive type antagonism. 5. PGI2 generation induced by the ionophore A23187 was unaffected by CV 6209 (up to 100 nmol/L) whereas basal PGI2 production by macrophages was reduced by lower concentrations (10-100 nmol/L). These observations are not consistent with a direct effect of CV 6209 on the enzymes involved in PGI2 synthesis but do suggest that endogenous Paf regulates basal PGI2 generation. 6. The non-competitive antagonism of guinea-pig macrophage Paf receptors gives further support to the contention that these receptors are distinct from those mediating aggregation of platelets and PMN. PMID- 2558828 TI - Evidence for a role of nitric oxide in the neurotransmitter system mediating relaxation of the rat anococcygeus muscle. AB - 1. The nitric oxide (NO) synthesis inhibitor NG-monomethyl-L-arginine (L-NMMA), but not D-NMMA, inhibited the NANC-mediated relaxations of the rat anococcygeus muscle, but did not affect the relaxation produced by sodium nitroprusside. 2. The inhibitory effect of L-NMMA was reversed by L-arginine but not by D-arginine, and prior exposure to L-arginine blocked the effect of L-NMMA. 3. The noradrenergically mediated contractions of the anococcygeus elicited by field stimulation were slightly enhanced by L-NMMA, but the response to noradrenaline was not affected. 4. The results suggest that NANC transmission in the rat anococcygeus muscle involves the generation of NO from arginine. PMID- 2558829 TI - A low molecular weight heparin ("fragmin") for routine hemodialysis: a crossover trial comparing three dose regimens with a standard regimen of commercial unfractionated heparin. AB - In 20 hemodialysis patients using mainly flat plate dialyzers, we have conducted a controlled randomized crossover trial of three dose regimens of a low molecular weight heparin (LMWH), "Fragmin" (Kabi 2165), in comparison with a standard dose regimen of commercial unfractionated heparin (UFH) that had previously been shown to provide effective anticoagulation. The aim of the present study was to find the lowest dosage regimen of the LMWH which would be as effective as the standard UFH regimen. The UFH regimen comprised a prime with heparinized saline, an initial intravenous bolus of 5,000 international units (IU) and an infusion of 1,500 IU/h. The three LMWH regimens comprised a LMWH-saline prime, an infusion of 750 anti-factor Xa (aXa) U/h and three different bolus doses: 1) LMWH-low: 3,000 aXa U. 2) LMWH-medium: 4,000 aXa U. 3) LMWH-high: 5,000 aXa U. With the UFH regimen, plasma heparin levels of around 1.0 IU/ml were maintained during the heparin infusion, declining to 0.71 IU/ml an hour after the infusion was terminated. The LMWH-medium regimen produced very similar plasma aXa levels. The LMWH-high regimen also produced similar plasma aXa levels: therefore, it had no advantage over the LMWH-medium regimen. The LMWH-low regimen produced significantly lower levels than the other regimens during the heparin infusion (0.81-0.85 aXa U/ml, p less than 0.025). Dialysis proceeded uneventfully at all times and plasma levels of fibrinopeptide A (FPA) were suppressed well with all 4 regimens (2.77-5.74 pmol/ml) but tended to rise after the infusion was switched off (5.52-8.45 pmol/ml).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558832 TI - [Aluminum deposition and Ca-hydroxyapatite formation in frontal cortex tissue of amyotrophic lateral sclerosis]. AB - To investigate a possible participatory role of an environmental metals in a pathogenetic process of amyotrophic lateral sclerosis (ALS), elementary and structural analyses of calcified substance in frontal cortex (percental gyrus) tissues of ALS patients were conducted using particle-induced X-ray emission (PIXE) and X-ray diffraction methods, as compared with control subjects. In the results of PIXE analysis, aluminum (Al) and calcium (Ca) contents in the frontal cortex tissues of the ALS patients were significantly increased with a high calcium (Ca)/phosphorus (P) Ca-hydroxyapatite (Ca-HAp). Although the content of peak ratio, as compared with those of control subjects. Only the Al content in the ALS patients showed significantly negative correlations with age at onset and/or at death. X-ray diffraction patterns from the sample (ashed at low temperature) of the frontal cortex tissues of the ALS patients were very similar to those from a standard sample of Ca-hydroxyapatite (Ca-HAp). Although the content of Ca-HAp in the mixed sample of frontal cortex tissues from the ten control subjects were not detectable (less than 3ppm), those of the ALS patients were markedly high, i.e., 111ppm in the mixed sample of five young patients with age under 60 and 168ppm in that of five old patients with age over 60. These results strongly indicated that Al may induce a calcifying degeneration in the CNS tissue of ALS patients, consequently leading to Ca-HAp formation as the final chemical compound. PMID- 2558831 TI - [Expression of nerve growth factor receptor in human benign peripheral nerve sheath tumor]. AB - We examined in vivo and in vitro expression of nerve growth factor (NGF) receptors in 13 dermal and 9 plexiform neurofibromas, 28 schwannomas, and 4 traumatic neuromas with an immunohistochemical method using a monoclonal antihuman NGF receptor antibody (ME20-4) and anti S-100 beta protein antibody. Immunoreactivity for NGF receptor and S-100 beta protein was universally observed on the principal cells of both neurofibroma and schwannoma in vivo. Moreover, we examined the NGF receptor and S-100 beta protein immunoreactivity in the culture system of the neurofibroma and schwannoma with an immunofluorescent double staining method. The principal cells of both tumors were positively stained by both ME20-4 and anti S-100 beta protein antibodies. These cells also express the ability for 125I-NGF binding, which was examined with an autoradiographic technique. These results suggest that NGF receptors are universally expressed on the S-100 beta protein positive Schwann-like cells composing of these tumors both in vivo and in vitro. In traumatic neuroma, however, a positive staining of NGF receptor was more strongly observed in the perineurium rather than in the endoneurial cells. PMID- 2558830 TI - [Ultrastructural immunocytochemical localization of electron-transport enzymes in mitochondrial myopathy]. AB - Ultrastructural localization of mitochondrial electron transport enzymes in biopsied muscle from a patient with mitochondrial myopathy was studied. We applied the immunoelectron microscopic technique using colloidal-gold labeled antibodies. The results demonstrated a very distinct and dense labeling by gold particles of complexes I, III, and IV in the inner mitochondrial membrane. The density of gold particles reacted to complex IV was cleanly decreased. This result was correlated with the decreased biochemical activity of complex IV. Labeling on the paracrystalline inclusions was substantially decreased in abnormal giant mitochondrial, but gold particles were extensively confined on the membrane of cristae surrounding the inclusions. PMID- 2558833 TI - [A case of Sjogren-Larsson syndrome with peripheral nerve involvement]. AB - A case of Sjogren-Larsson syndrome (SLS) with peripheral nerve involvement was described. A patient was a 37-year-old man with marked mental retardation who had ichthyosis since several months of age. He developed spastic gait at age 20 and noticed muscle weakness and atrophy in lower extremities at age 30. On examination, electromyogram showed neurogenic changes with polyphasic motor unit potentials of high amplitude and long duration, and motor and sensory conduction velocities were slightly decreased in lower extremities. Muscle biopsy specimens of the left quadriceps showed grouped atrophy and pyknotic nuclear clumps, suggestive of neurogenic changes. Left sural nerve biopsy showed decreased number of myelinated fibers. On electron microscopy, some myelinated fibers revealed proliferation of the organelles in axons with disruption of the myelin sheath, and glycogen granules in Schwann cell cytoplasm. Unmyelinated fibers showed no reduction in the density, but accompanied numerous collagen pockets and Schwann cell cytoplasmic processes arranged in many stacks, suggesting the presence of some degenerative changes. With the present findings at peripheral level and a review of the literature, we may assume that SLS has extensive disorders of the ectodermal tissues including the peripheral nerve as well as the skin and the central nervous system. PMID- 2558834 TI - Evaluation of immunofluorescent reagents, centrifugation, and conventional cultures for the diagnosis of adenovirus infection. AB - In this study we have evaluated four fluorescent antibody reagents, three monoclonal and one polyclonal, for identification of adenovirus isolates and compared four conventional cultures (human embryonic kidney, A549, HEp-2, and MRC 5 cells) with centrifugation culture for rapid diagnosis. For identification of adenovirus isolates by immunofluorescence, CDC reagent and Adenoclone, both monoclonal antibodies to the hexon group-reactive antigen, were more sensitive and easier to interpret than the other two reagents tested. HEK and A549 cells were the most sensitive for isolation of adenovirus. Although A549 cells were an inexpensive alternative to HEK, A549 cell monolayers deteriorated more rapidly and passages were more often required. Centrifugation cultures with A549 cells detected 77% of positives within 2 days and 100% within 5 days, whereas isolation in conventional culture required up to 10 days for HEK and up to 20 days for MRC 5 cells. PMID- 2558835 TI - [The effect of a single dose of dexamethasone on the production of superoxide anion and on the chemotaxis of blood polynuclear neutrophils in the horse]. AB - Chemotaxis and superoxide anion production of neutrophils in healthy horses were investigated before and 8 h after, a single injection of dexamethasone at a dose of 0.045 mg/kg. Chemotaxis was studied by the technique of migration under agarose and superoxyde production was measured by ferricytochrome c reduction. Superoxide anion production was not changed, but the chemotactic index, with zymosan activated horse serum, was increased. The FMLP chemoattractant effect, at 10(-4) M, was slightly enhanced. PMID- 2558836 TI - Detergent solubilised bovine viral diarrhea virus elicits a similar immune response as the inactivated virus. AB - The immune response to two detergent solubilised preparations of Bovine Viral Diarrhea Virus (BVDV) proteins were compared with the response to a chemically inactivated BVDV preparation. The antibody titres were measured with the standard immunofluorescence assay and with a newly developed enzyme linked immunosorbent assay (ELISA). The range of viral protein specificity of the antibodies was compared using SDS-polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blot analysis. The results clearly demonstrate that the detergent solubilised viral proteins are equally as effective immunogens as the killed BVDV vaccine both in terms of antibody titers and specificity. PMID- 2558837 TI - An enzyme-linked immunosorbent assay using canine coronavirus-infected CRFK cells as antigen for detection of anti-coronavirus antibody in cat. AB - From the reasons that canine coronavirus (CCV) grows more efficiently than feline coronavirus in a cell culture and they are mutually related in their antigenicities, an enzyme-linked immunosorbent assay (ELISA) using CCV-infected feline kidney (CRFK) cells as substrate antigens was developed for detection of anti-coronavirus antibodies in cats. It was indispensable for generating coronavirus-specific ELISA antibody activities that the sample was applied to the mock-infected, normal CRFK cells in parallel with the CCV-infected cells and then the optical density values given by the mock-infected cell antigen were subtracted from those given by the virus-infected cell antigen. On the basis of ELISA antibody titers obtained in sera from the cats experimentally infected with CCV and from the spontaneous feline infectious peritonitis (FIP) cases, the ELISA described in the present study was found to be applicable as a simple and easy serologic test which was able to detect anti-coronavirus antibodies as efficiently as the indirect immunofluorescence assay with homologous FIP virus. PMID- 2558838 TI - Detection of rotavirus in faecal specimens with a monoclonal antibody enzyme linked immunosorbent assay: comparison with polyclonal antibody enzyme immuno assays and a latex agglutination test. AB - Monoclonal antibodies have been produced against the 81/36F strain of rotavirus. One of them, was chosen as diagnostic reagent: it showed high ELISA reactivity with all the bovine, human and porcine rotavirus strains tested and reacted with VP6, structural protein product known to support the common rotavirus antigen. A sandwich ELISA procedure using the chosen monoclonal as "capture and detecting" antibody was performed to detect rotavirus in faecal samples from experimentally inoculated newborn calves: it always gave a negative response with meconium and a positive response for the stool specimens which rotavirus have been isolated. This assay was compared with Enzygnost and Slidex Rota Kit tests and with a non commercial sandwich ELISA test using polyclonal antibodies: it showed more sensitivity than the agglutination test and was as sensitive as the other two tests to detect rotavirus in routine diagnostic material. The test evaluated showed no equivocal results. PMID- 2558839 TI - Debut of a somatostatin analog: octreotide in review. AB - Octreotide is a long-acting cyclic octapeptide with pharmacologic actions mimicking those of the natural hormone somatostatin. It can suppress the secretion of serotonin, as well as the gastroenteropancreatic peptides gastrin, vasoactive intestinal peptide (VIP), insulin, glucagon, secretin, motilin, and pancreatic polypeptide. It also suppresses growth hormone and decreases splanchnic blood flow. Octreotide is completely and rapidly absorbed following subcutaneous injection and has an elimination half-life of 1.5 hours. Clinical trials reviewed here show octreotide useful in the treatment of diarrhea associated with VIP secreting tumors, as well as diarrhea and flushing associated with carcinoid syndrome, both conditions for which the drug is approved. Clinical trials involving the use of octreotide in the treatment of acromegaly are also reviewed. Adverse reactions to octreotide are mild to moderate and most commonly involve injection site pain and diarrhea. Drug interactions are apparently related to the drug's pharmacologic effects. Octreotide is given subcutaneously two to three times daily, with daily doses ranging from 50mcg to 1,500mcg per day. Further research appears necessary to clarify dosing issues. PMID- 2558840 TI - Functional and molecular changes in colony stimulating factor secretion by osteoblasts. AB - Two populations of primary osteoblasts and the cloned murine osteoblast cell line MC3T3 were activated with osteotropic agents and the conditioned media tested for the presence of macrophage colony stimulating factor (M-CSF). Differences in ability of the three populations to secrete the cytokine as well as in the efficacy of the activating agents used to induce it were observed. The ability of these agents to modulate M-CSF mRNA levels was also examined. The data indicate that osteoblasts do not respond uniformly and the reasons for this are discussed. Osteoblasts may exert major regulatory influences on hematopoiesis via CSF secretion as well as function in their more traditional role as the cells responsible for new bone formation. PMID- 2558841 TI - Hormonal regulation of collagen gene expression in osteoblastic cell-overview and new findings. AB - Type I collagen synthesis in osteoblasts is regulated by a variety of systemic hormones, growth factors and locally produced factors. With the use of cDNA probes and hybridization analysis, it appears that many of these agents alter collagen synthesis by a pretranslational mechanism. PMID- 2558842 TI - Methoxatin (PQQ), coenzyme for copper-dependent amine and mixed-function oxidation in mammalian tissues. AB - The newly discovered coenzyme PQQ can now be measured at picomole levels with redox cycling methods developed in our laboratory. PQQ-peptides have been obtained from digests of the copper quinoenzymes, diamine oxidase, lysyl oxidase and dopamine-beta-hydroxylase. PQQ is present in egg yolk and milk, suggesting its immediate availability for developing embryos and newborn animals. We suggest that PQQ, when exposed in traumatized, ischemic, inflammed or pathological tissues, may catalyze the formation of large amounts of superoxide and should be considered as a source of oxidative stress when planning pharmacotherapeutic intervention. PQQ and quinoproteins play a role in the redox metabolism and structural integrity of cells and tissues. PMID- 2558843 TI - Transforming growth factor-beta regulation of collagenase, 72 kDa-progelatinase, TIMP and PAI-1 expression in rat bone cell populations and human fibroblasts. AB - Quiescent cultures of normal fetal rat calvarial bone cell populations (RC I and RC IV) and human fibroblasts were incubated with 1.0 ng/ml TGF-beta and the conditioned culture media were processed individually to separate collagenase and 72 kDa-progelatinase from TIMP, the tissue inhibitor of matrix metalloendoproteinases, using mini-columns of heparin- and gelatin-Sepharose. Collagenase synthesis was decreased progressively by TGF-beta in fibroblasts despite a 1.6-fold increase in secreted protein levels and a approximately 1.8 fold increase in 72 kDa-progelatinase synthesis. The human fibroblasts and the osteoblast-enriched RC IV cells showed a greater TGF-beta-induced stimulation in 72 kDa-progelatinase levels over controls compared with the RC I cells. In contrast to RC IV cells, in which TIMP mRNA levels were increased 2.9-fold by TGF beta, the constitutive level of TIMP transcripts in the RC I cells was greater than 20-fold over that of the RC IV cells, but was not elevated by TGF-beta. TGF beta also increased TIMP expression in fibroblasts approximately 1.7-fold and PAI 1 levels approximately 5-fold in RC IV cells, and greater than 10-fold in fibroblasts. PMID- 2558844 TI - Morphofunctional examination of the heart in rats with the alcohol withdrawal syndrome. AB - Using histochemical methods, light and electron microscopy, authors examined rat heart 2-6 hours, 1, 3, and 7 days after discontinuation of forced intoxication with alcohol. At the same time, they assessed the contractile function and creatine phosphokinase (CPK) activity in the isolated perfused heart, and the development of animal destruction. Ethanol withdrawal was followed by escalation of vascular disorders in the heart, dystrophic changes in the subcellular structures, considerable polymorphism in enzyme distribution and activity, and formation of foci containing disintegrating myocytes with contractures. The contractile function was impaired and CPK release increased in the isolated heart. The changes were most marked 3 days after ethanol discontinuation to disappear after 7 days. Two to seven days after ethanol cessation, 13.1% of rats perished. Cardiac injury due to alcohol withdrawal syndrome may be one of the factors leading to the development of alcohol cardiomyopathy and a cause of sudden death in patients with documented alcohol abuse. PMID- 2558845 TI - Prefeeding of aldose reductase inhibitor and galactose cataractogenesis. AB - Our recent investigations have shown that the Eisai compound, E-0722, (2R-4S-6 fluoro-1-2-methylspirochroman 4,4'-imidazolidine 2,5'-dione) is a more potent aldose reductase inhibitor than Sorbinil (D-6-fluorospirochroman 4,4' imidazolidine 2,5'-dione). In the previous studies these aldose reductase inhibitors were added to the 50% galactose diet fed to rats to determine their effect on galactose-induced alterations in the lens and the development of cataract. In this report we present our results on the effect of prefeeding the aldose reductase inhibitor, E-0722, on the alterations in rat lens following subsequent feeding of galactose. For this study, young Sprague Dawley rats were prefed either rat chow or rat chow plus 50% galactose containing 1mg/day/Kg body weight of E-0722 for 1 or 2 weeks. After this dietary regimen, the animals were transferred to diets containing 50% galactose for different periods. For controls, rats were fed either rat chow or 50% galactose without the prefeeding of E-0722. Our results obtained through gross observation of the lenses, light microscopic studies of lens sections and assay of Na+-K+-ATPase (NPPase) activity show that the prefeeding of E-0722 prior to galactose feeding delays galactose induced alterations and the development of mature cataract. PMID- 2558846 TI - Coupling of 22Na and 36Cl uptake in cultured pigmented ciliary epithelial cells: a proposed role for the isoenzymes of carbonic anhydrase. AB - Uptake studies with 22Na and 36Cl were performed in cultured bovine pigmented ciliary epithelial cells (PE) to investigate interdependence of Na+ and Cl- transport. (1) 22Na uptake into NaCl depleted cells was stimulated by Cl-. This stimulation was abolished by the simultaneous application of amiloride (1 mM) and bumetanide (0.1 mM), indicating two independent mechanism for Cl- stimulated Na+ uptake: loop diuretic sensitive Na+/Cl- symport and an indirect stimulation of Na+/H+ exchange by Cl-. The latter component of Cl- stimulated Na+ uptake was HCO3- dependent. (2) 36Cl uptake was increased by extracellular Na+. Na+ stimulated Cl- uptake also consisted of two components. One was bumetanide sensitive and the other was blockable by amiloride and partly inhibited by the carbonic anhydrase (CA) inhibitor methazolamide (0.1 mM). (3) Homogenized PE cells were tested for biochemical CA activity using an electrometric method. The cytoplasmic as well as the membrane fraction contained specific CA activity. (4) A model is presented for Na+ and Cl- transport into PE: in addition to Na+/Cl- symport, Na+/H+ and Cl-/HCO3- double exchange may operate in the ciliary epithelium. The latter mechanism provides NaCl uptake into the cell in exchange for H+ and HCO3-, which recycle as CO2 across the membrane. This recycling of CO2 and HCO3-/H+ (and hence indirectly NaCl uptake) is facilitated by the cooperation between membrane bound and cytoplasmic CA. PMID- 2558848 TI - Histopathologic study of herpes virus-induced retinitis in athymic BALB/c mice: evidence for an immunopathogenic process. AB - In order to determine whether antiviral immunity is pathogenic in mouse eyes, HSV 1 was injected into the anterior chamber of one eye of adult athymic BALB/c mice. The eyes of these T cell deficient mice were examined clinically and histopathologically for ocular disease. The anterior segment of injected eyes developed progressive inflammatory reactions that eventually destroyed the ciliary body and then progressed to the posterior compartment where partial necrosis occurred, but only in the inner layers of the retina. A milder form of the same process developed between 7 and 10 days in the contralateral eye. Uninoculated eyes displayed little evidence of choroiditis, hemorrhage, massive necrosis, or disintegration of the architecture of the retina. Since these are features that are found in contralateral retinas of euthymic BALB/c mice infected in one eye via the anterior chamber route, it is concluded that acute retinitis found in contralateral eyes of immunocompetent mice has an immunopathogenic basis. However since euthymic mice develop anterior chamber associated immune deviation (ACAID) (and therefore do not display virus-specific delayed hypersensitivity), the identity of the relevant immune effector remains unknown. Based on these observations and our previous ocular findings following intracameral inoculation of HSV-2, we suggest that in susceptible mice, herpes simplex viruses can induce several pathogenetically distinct forms of retinitis, some of which are mediated by virus-specific immune effector cells. PMID- 2558847 TI - Experimental parameters affecting the ultraviolet-induced tryptophan modification in the lens. AB - Experimental parameters affecting photolysis of tryptophan in the lens were evaluated, including: (a) the use of previously-frozen vs. freshly excised lenses, (b) the use of lenses in aqueous buffered solutions vs. in air, (c) the species source of the lens, and (d) the effects of oxygen depletion. Lenses were subjected to monochromatic ultraviolet radiation followed by monitoring of the tryptophan fluorescence spectra while exciting at two or more wavelengths. Previously frozen lenses showed faster UV-induced degradation of tryptophan, particularly in aqueous media. A rapid leakage of fluorescent components (including amino acids and peptides) from intact lenses was observed. Evidence for the artifactual production of free radicals during photometric monitoring of samples in solution was obtained. Species dependent differences were also observed. PMID- 2558849 TI - An improved model of recurrent herpetic eye disease in mice. AB - Mice were passively immunized with serum containing antibodies to herpes simplex virus type 1 (HSV-1) before inoculation on the cornea with HSV-1 strain McKrae. After such immunization most mice survived and most had normal eyes. When primary infection had subsided, mice with normal eyes were selected and treated with cyclophosphamide, dexamethasone and UV irradiation of the inoculated eye or UV irradiation alone, to reactivate latent virus. After either treatment mice developed signs of recurrent infection (virus in eyewashings and recurrent corneal and/or lid disease). The incidence of such signs was 17/33 (52%) in mice receiving immunosuppressive drugs and UV irradiation and 19/32 (59%) in mice given UV irradiation alone. In mice treated with either stimulus dendritic or geographic ulceration of the cornea was seen. These closely resembled the herpetic lesions seen in humans. There was good correlation between the pattern and distribution of recurrent corneal disease and the distribution of cells containing virus antigens in corneal epithelial sheets. Again, as in humans, the induction of recurrent infection was found to correlate poorly with a rise in the level of serum neutralizing antibody. In mice treated with UV irradiation alone corneal ulcers healed and the eyes returned to normal. By contrast, in mice given immunosuppressive drugs and UV irradiation, the ulceration became more severe and the eyes became opaque and vascularized. The use of passive immunization has greatly improved our previously reported model of recurrent herpetic eye disease since it has increased the incidence of mice suitable for the induction of recurrent infection and has increased the incidence of such infection. PMID- 2558850 TI - VIP modulation of cultured glial cells of the rat retina. AB - Cultured retinal glial cells from the rat are responsive to modulation by vasoactive intestinal peptide (VIP). VIP (1 X 10(-6) M) elevated the intracellular cyclic AMP concentration from the basal level of (4.4-11.1) p mole/mg protein to (354-440) p mole/mg protein in three minutes at 25 degrees C. The half-maximal concentration is 4.8 X 10(-8) M, which is similar to that observed in the cultured retinal glial cells from the chick embryo. PMID- 2558851 TI - Short-term desensitization of prostaglandin F2 alpha receptors increases cyclic AMP formation and reduces inositol phosphates accumulation and contraction in the bovine iris sphincter. AB - The effect of short-term prostaglandin (PG) desensitization on PGF2 alpha receptor-mediated inositol phosphates accumulation, 1,2-diacylglycerol production, measured as phosphatidic acid (PA), myosin light chain (MLC) phosphorylation, cAMP formation and contraction was investigated in bovine iris sphincter smooth muscle. We have found that incubation of the sphincter with 25 microM PGF2 alpha for 45 min leads to: (a) significant loss in sensitivity of the tissue to PGF2 alpha receptor-stimulated inositol phosphates accumulation, PA production, MLC phosphorylation and contraction, and (b) significant increase in both basal and PGF2 alpha-stimulated cAMP formation. These changes are probably not due to reduction in phospholipid synthesis because there were no detectable differences in basal phospholipid labeling, either from 3H-inositol or from 32P, between normal and desensitized muscles. Preincubation of the sphincter in the absence of PGF2 alpha for 45 min did not lead to alterations in the biochemical pharmacological responsiveness of the control muscle to PGF2 alpha. Our results suggest that desensitization of PG receptors in the iris sphincter occurs by a receptor-specific process. The PG receptor mediating contraction (IP3-Ca2+) is selectively susceptible to desensitization, in contrast with the receptor mediating smooth muscle relaxation (cAMP). These findings add further support to the developing hypothesis that there are functional and biochemical reciprocal interactions between the IP3-Ca2+ and cAMP messenger systems in the iris of the mammalian eye. PMID- 2558852 TI - Elephantiasis-like cutaneous metastases from signet-ring cell carcinoma of the stomach. AB - We report the clinical features and course of a 66-year-old patient with gastric carcinoma, in whom multiple cutaneous metastases occurred one year after he underwent subtotal gastrectomy. Although internal malignancies rarely metastasize to the skin, when they do this may have profound diagnostic, therapeutic, and prognostic significance. Cutaneous metastases are usually synonymous with a rapidly progressive disease and a poor prognosis. This case, in which cutaneous metastases were found in the lower limbs, is notable since usually less than 4 percent of cutaneous metastatic lesions are found in this region. Moreover, the case seems unique because of the predominating dermiotrophic metastases, with only minor internal involvement. PMID- 2558853 TI - Localization of the gene encoding the alpha-subunit of the acetylcholine receptor on chromosome 2 of the mouse. PMID- 2558854 TI - Refined mapping of the three DNA repair genes, ERCC1, ERCC2, and XRCC1, on human chromosome 19. AB - Three DNA repair genes, ERCC1, ERCC2, and XRCC1, have been regionally mapped on human chromosome 19. ERCC2 and XRCC1 have been assigned to bands q13.2----q13.3 by in situ hybridization using fluorescently-labeled cosmid probes. ERCC1 and ERCC2 have been found to be separated by less than 250 kb by large fragment restriction enzyme site mapping. PMID- 2558855 TI - Human differentiation-stimulating factor (leukemia inhibitory factor, human interleukin DA) gene maps distal to the Ewing sarcoma breakpoint on 22q. AB - The human gene encoding differentiation-stimulating factor (D-factor) has previously been isolated and shown to be identical to leukemia inhibitory factor (LIF). We have determined a fine structure map of approximately 20-kb surrounding the D-factor/LIF gene. Southern blot analysis using a somatic cell hybrid panel shows that the gene maps to chromosome 22. D-factor/LIF was further sublocalized to 22q11.2----q13.1, distal to a Ewing sarcoma (ES) breakpoint, using a second somatic cell hybrid panel. Probes to the 5' and 3' regions of the locus and the cDNA were used to screen for restriction fragment length polymorphisms, but none were detected. Analysis by pulsed field gel electrophoresis suggests that D factor/LIF is not near the ES breakpoint. PMID- 2558856 TI - The human mineralocorticoid receptor gene (MLR) is located on chromosome 4 at q31.2. AB - The gene for the human mineralocorticoid receptor (MLR) was previously localized to chromosome 4. Here, we have localized this gene to 4q31.2 by in situ hybridization. This precise mapping of MLR will assist in the linkage analysis and genetic characterization of pseudohypoaldosteronism, an autosomal recessive disorder which likely results from a defect in the MLR gene. PMID- 2558857 TI - A somatic cell hybrid panel to facilitate identification of DNA sequences in the vicinity of the incontinentia pigmenti locus (IP1). AB - Somatic cell hybrids that retain derivative X chromosomes from women with sporadic incontinentia pigmenti (IP1) and de novo X/autosomal translocations with consistent breakpoints at Xp11.21 were constructed. An assembled hybrid panel was used to physically map DNA sequences in relationship to the IP breakpoint. DSX14 was found to map to region Xp11.21----p11.1. Regional assignments of 19 X chromosomal loci were reviewed. PMID- 2558858 TI - [Role of local flaps in the treatment of interdigital commissure contractures]. AB - The authors present a review of the most reliable and effective local flaps in the treatment of moderate web contractures. In their opinion, they are very helpful in that pathology's treatment provided that indications and limits are well pointed out. Advantages and disadvantages of each described procedure reported. PMID- 2558859 TI - [Reconstruction of the first interdigital commissure with distal flaps]. AB - Authors report some examples of distant flaps which they judge excellent in first web reconstruction following severe posttraumatic skin retraction. The utilization of these procedures is suggested in those cases, particularly if complicated by skin loss, which need fairly large amount of skin with thin subcutaneous tissue like the one of the three described flaps. PMID- 2558860 TI - Anti-topoisomerase II recognizes meiotic chromosome cores. AB - At meiotic prophase the chromatin becomes arranged in loops on newly formed chromosome cores. The cores of homologous chromosomes become aligned in parallel and thus form the synaptonemal complex (SC), a structure found in the meiocytes of nearly all recombinationally competent, sexually reproducing organisms. We report that two polyclonal antibodies against topoisomerase II (topo II), which recognize the mitotic metaphase chromosome scaffold give, at pachytene, a positive immunocytological reaction with the chromatin and, predominantly, with the cores and centromeric regions of the paired chromosomes. It therefore appears that during meiotic prophase, topo II - a DNA-binding enzyme implicated in transient double-strand breaks, chromosome condensation, and anaphase separation is associated with the chromatin and SCs of the pachytene and diplotene chromosomes. PMID- 2558861 TI - The value of fine-needle aspiration biopsy in the cytodiagnosis of salivary gland lesions. AB - Fine-needle aspiration cytology (FNAC) was performed on 195 cases of salivary gland lesions. The smears were technically adequate in 178 cases. Tissue examination was available for subsequent histocytologic correlation in 57 cases. The cytodiagnosis included inflammatory lesions (59) and benign (68) and malignant (51) tumors (total, 119). The accuracy of cytodiagnosis was 87.7% with a sensitivity of 80.9% and a specificity of 94.3%. Exact histologic typing was possible in 61.9% of the malignant tumors. Mucoepidermoid tumors and cellular atypical pleomorphic adenoma posed difficulties in cytodiagnosis. PMID- 2558862 TI - Cytology of primary versus secondary genital herpesvirus infection. AB - Certain authors state that primary genital herpes infections mainly produce cells with ground-glass nuclei, whereas secondary infections produce a higher percentage of multinucleated or single cells with prominent intranuclear inclusions. The cervicovaginal smears of 11 patients with primary genital herpes infection and 16 patients with secondary infection were studied to determine the validity of this differentiation. The percentage per patient of virally infected cells containing distinct inclusions ranged from 0% to 35% in the primary herpes group and 0% to 100% in the secondary herpes group. In eight additional patients, the initial herpes simplex virus (HSV)-positive smear was compared with recurrent positive smears taken from 2 to 120 mo later. No statistical difference in the incidence of distinct inclusions in the infected cells was detected. Finally, primary rabbit kidney and the heteroploid A549 cell cultures were inoculated with herpes viruses and followed by fixation with several agents at various time intervals. Alcohol and Bouin's fixative favored the recognition of distinct inclusions, as did the duration of the infection. PMID- 2558863 TI - Ductal carcinoma-in-situ of the breast: fine-needle aspiration cytology of 12 cases. AB - Aspiration specimens from 12 patients with histologically documented ductal carcinoma-in-situ (DCIS) of the breast (seven patients) or DCIS with minute foci of stromal invasion (five) were evaluated. Five patients presented with palpable masses, 1.5-4.0 cm, and four patients presented with localized thickening, associated with nipple erosion and discharge in two of them. One patient had nipple inversion, and one patient had bilateral nipple discharge. In one patient, no apparent abnormality of the breast was present. Mammography was either suspicious for or strongly suggestive of carcinoma in 10 patients and negative in two. Aspirates from all patients were composed of fragments of atypical ductal epithelium and numerous single epithelial cells. In nine cases, the smears were hypercellular and similar to aspirates of typical invasive ductal carcinoma. Calcifications were present in six cases. In four of these, associated tumor necrosis was evident. Cytologic features separating DCIS patients from those showing minimal stromal invasion or common types of invasive ductal carcinoma (IDC) were not identified. We conclude that fine-needle aspiration cytology of DCIS is identical to that of IDC. If preoperative radiotherapy or chemotherapy is considered in the management of invasive breast carcinoma, cutting-needle biopsy for confirmation of tumor invasion is necessary. PMID- 2558866 TI - Superpolylinkers in cloning and expression vectors. AB - Versatile DNA polylinkers of more than 300 bp were constructed. They contain the recognition sequences of all restriction enzymes--whether known or still to be discovered--that recognize palindromic hexamers. In addition to these 64 uninterrupted hexameric recognition sites, a number of sites containing interrupted palindromes and nonpalindromic sequences and two recognition sequences with 8 bp are present. Polylinkers (in several variants) were inserted into frequently utilized Escherichia coli cloning vectors such as pBluescript (yielding pSLJ10, pSL250, pSL260, pSL270, and pSL300), pUC18/pUC19 (yielding pSL180 and pSL190, respectively), or pUC118/pUC119 (yielding pSL1180 and pSL1190, respectively). A subtle color discrimination between presence and absence of insert in pSL300 (mid-blue to light-blue or white) was seen in a number of test ligations. The mid-blue color that is generated by pSL300 is presumably due to translational restarts. A different intergenic region for translational restarts was used in plasmids pSL251, pSL261, pSL271, and pSL301. The polylinker was also inserted into expression vector pUC120, yielding pSE1200, and into expression vector pKK233-2, yielding pSE220 and a shortened version thereof, pSE280. Finally, the polylinker was inserted into pTrc99A, resulting in pSE380, which carries a lac repressor gene. This expands the use of the expression system beyond lacIq strains to other bacterial hosts. These versatile vectors have broad applications in genetic engineering. PMID- 2558864 TI - Alkali therapy of diabetic ketoacidosis: biochemical, physiologic, and clinical perspectives. PMID- 2558865 TI - Mapping contacts between unpurified human progesterone receptor and the hormone response element of mouse mammary tumor virus. AB - Binding of steroid hormone receptors to specific recognition sites of hormone inducible genes is one of the events required for hormonal regulation of gene transcription. We have employed an immunoprecipitation assay to map the interaction between unpurified human progesterone receptors from crude nuclear extracts of T47D cells and the hormone response element of the mouse mammary tumor virus (MMTV). DNase I footprints and methylation interference patterns are similar to those reported with highly purified rabbit progesterone receptors, suggesting that both human and rabbit receptors recognize similar features in the hormone response element. More importantly, these patterns suggest that if other factors are associated with unpurified nuclear receptor, they do not alter the contacts made by receptor nor do they make contacts themselves with MMTV DNA in a manner detected by DNase I or methylation interference assays. The sites of interaction of receptors bound with the clinically important progestin antagonist, RU 486, are comparable to those observed with an agonist-receptor complex. These results suggest that the antagonist prevents receptor action at a step after its recognition and binding to specific sites on a hormone-responsive enhancer element. PMID- 2558867 TI - Alkaline phosphatase fusions to the respiratory syncytial virus F protein as an approach to analyze its membrane topology. AB - Manoil and Beckwith (1985) have constructed a transposon, TnphoA, that permits the generation of hybrid proteins composed of alkaline phosphatase (AP) lacking its signal peptide fused to amino-terminal sequences of other proteins. This transposon has been used to localize export signals and analyze membrane topology of bacterial proteins. We have applied this approach to the membrane fusion protein (F) of respiratory syncytial virus (RSV). The transposon TnphoA and a plasmid directing bacterial expression of the F gene were used to construct F-AP hybrids. These hybrids yielded AP activity, indicating the presence of viral sequences that promoted protein transport through the cytoplasmic membrane. Sequence analysis showed that TnphoA was inserted at four different positions within the F1 subunit. Deletion of the hydrophobic F1 amino-terminus (fusion related domain) resulted in AP transport to the periplasm, suggesting that the hydrophobic amino-terminus of the F2 subunit is sufficient to promote protein export. Some hybrids were apparently cleaved at or near the F2/F1 junction. The periplasmic localization of an uncleaved hybrid strongly suggested that the fusion-related domain of the F protein, when in the uncleaved F0 precursor, can be moved across the bacterial cytoplasmic membrane. Although these results apply to the recombinant F protein, they agree with the presumed signal sequence and membrane topology of the native F glycoprotein. Thus, this method may be useful in determining membrane topology and in localizing important domains of viral proteins. PMID- 2558868 TI - Isolation and sequence of a cDNA clone for human calcineurin B, the Ca2+-binding subunit of the Ca2+/calmodulin-stimulated protein phosphatase. AB - We have identified and cloned human cDNA for the Ca2+-binding subunit of calcineurin, the brain isozyme of the Ca2+/calmodulin-stimulated protein phosphatase. The 2.5-kb cDNA has an open reading frame of 510 bp, a leader sequence of at least 500 bp, and a 1,277-bp 3'-noncoding sequence. The deduced sequence of the human protein differs from bovine brain calcineurin B by an additional valine at the carboxyl terminus and substitution of Met-11 and Ser-153 by cysteine. A partial clone of the mouse protein corresponding to amino acids 75 150 was also isolated. This portion of the human and mouse protein sequence is identical, with the DNA sequences showing 94% identity. The respective mRNAs in human and mouse are also of similar size. As was observed with protein levels, mRNA abundance in brain is 20-60 times that found in other tissues with the exception of HeLa cells which, like brain, contain abundant calcineurin B mRNA. PMID- 2558869 TI - [Monoclonal antibodies against gastric cancer and their selective reaction on various tissues]. AB - Spleen cells from Balb/c mice immunized in sequence with five human gastric cancer cell lines were fused with murine myeloma cell line SP2/0. Hybridomas 3F4, 3G9 and 3H11 secreting monoclonal antibodies (mAb) against gastric cancer were obtained through selective culture and screening. These mAb produced by the immunization procedure have good selectivity and high positive rate in reaction on gastric cancer. The positive rate of reaction on gastric cancer cells and tissues could reach 5/5 and 84.8-93.5%, respectively, whereas there was almost no positive reaction on normal cells and tissues As there was no correlation between the positive reaction of gastric cancer and their histopathologic typing, and the cross reaction of mAb with other tumors and fetal gastrointestinal tissues was quite high, the corresponding antigens of these mAb were considered as extensive oncofetal antigens. PMID- 2558870 TI - [Paget's disease of the nipple]. AB - Fifty-five female patients with Paget's disease of the nipple treated in our Hospital from 1958 to 1987 are reported. It comprised 1.1% of all the breast carcinomas. The lesions was on the left side in 27 and on the right side in 28. The average age was 48 years. The primary lesion was in breast ducts and then invaded the nipple, areola superficially and deeper breast tissue. Dermal manifestation of Paget's disease is easily confused with chronic dermopathy. In this series, four patients (7.2%) did not have the typical symptoms so pathologic or cytologic examination was relied upon for correct diagnosis. Thirty-one patients were treated by radical mastectomy giving a 3-year disease-free rate of 70.9%. It seems that radical mastectomy is more superior to the other surgical procedures. The 3-year recurrent rate was 38% for patients with Paget's disease of the nipple coexisting with breast carcinoma, but no recurrence was found in patients with simple Paget's disease of the nipple. The 3-year survival rate was 46.4% in patients with axillary lymph node metastasis, 85.2% in patients without, 54.3% in patients with palpable masses in the breast and 85.0% in patients without. Paget's disease of the nipple coexisting with breast carcinoma, axillary lymph node metastasis, palpable breast masses has poor prognosis. PMID- 2558871 TI - [Preoperative intensive chemotherapy of operable breast cancer]. AB - Eighty-one patients with operable breast cancers received the short-term and intensive preoperative chemotherapy, followed by radical mastectomy are presented. The total response rate in the primary tumor to chemotherapy was 59%. Seven patients (8.6%) had complete remission, 41 (50.6%) partial remission and 33 (40.7%) stable lesions. No cancer cells were found histopathologically by serial sections from two primary lesions after chemotherapy. The authors found that tumor with small size, short course and regular shape had a better response rate. Histopathological type, firmness and boundary of tumor, menopausal status, and estrogen receptor (ER) status were not related to the chemotherapeutic effect. No obvious toxicity and side effects were found during the chemotherapy. The chemotherapy did not influence the wound healing. The results indicate that the chemotherapy has a favorable effect on breast cancer and is safe and valid. The response rate of breast cancer to chemotherapy before operation can be used as breast cancer chemosensitivity test in vivo. PMID- 2558872 TI - [Possible participation of specific receptors in the transport of nucleic acids in cells]. PMID- 2558873 TI - PDGF receptors on cells of the oligodendrocyte-type-2 astrocyte (O-2A) cell lineage. AB - It has been shown previously that cultures of rat optic nerve contain three types of macroglial cells--oligodendrocytes and two types of astrocytes. Type-1 astrocytes develop from their own precursor cells beginning before birth, while oligodendrocytes and type-2 astrocytes develop postnatally from a common bipotential precursor called the O-2A progenitor cell. Proliferating O-2A progenitor cells give rise to postmitotic oligodendrocytes beginning around birth, and to type-2 astrocytes beginning in the second postnatal week. Studies in vitro have suggested that platelet-derived growth factor (PDGF), secreted by type-1 astrocytes, plays an important part in timing oligodendrocyte development: PDGF seems to keep O-2A progenitor cells proliferating until an intrinsic clock in the progenitor cells initiates the process leading to oligodendrocyte differentiation. The clock apparently determines when a progenitor cell becomes unresponsive to PDGF, at which point the cell stops dividing and, as a consequence, automatically differentiates into an oligodendrocyte. Here we have used radiolabelled PDGF to show that O-2A progenitor cells have PDGF receptors, suggesting that these cells respond directly to PDGF. The receptors resemble the type A PDGF receptor previously described on human fibroblasts and are initially retained when progenitor cells stop dividing and develop in vitro into oligodendrocytes. The latter finding indicates that receptor loss is not the reason that progenitor cells initially become mitotically unresponsive to PDGF. PMID- 2558874 TI - Fasting insulin levels in normal Kenyan Africans and a case report on insulinoma. AB - The mean fasting insulin level in 30 apparently normal Kenyan Africans was determined by Radio-Immuno Assay (RIA) technique based on coat A-count. The mean value was found as 16.33 microIu/ml with a standard deviation (s.d) of 10.51 microIu/ml. The mean at 95% confidence interval (CI) for the population studied was 16.33 +/- 1.92. The findings are similar to what have been reported among the caucasians. This paper also reports on a case of insulinoma in a young female patient at Kenyatta National Hospital (KNH). PMID- 2558875 TI - Interaction of antiaggregant molecule ajoene with membranes. An ESR and 1H, 2H, 31P-NMR study. AB - The structure of ajoene, a molecule extracted from garlic, has been studied by 1H NMR and its interaction with model membranes by 1H-, 2H-, 31-P-NMR and ESR experiments. This study clearly shows that the ajoene molecule is located deep in the layer and is close to the interlayer medium. Moreover while NMR experiments show that the membrane structure is only slightly affected by the presence of ajoene, ESR experiments reveal significant modifications in phospholipid dynamics. This interaction, observed before with the phenothiazine derivative, promazine, results in an increase of the membrane fluidity in its hydrophobic part and could be related to clinical properties of ajoene. PMID- 2558876 TI - Catecholaminergic modulation of corticotropin-releasing factor and adrenocorticotropin secretion. PMID- 2558877 TI - Parathyroid hormone-like proteins: biochemical responses and receptor interactions. PMID- 2558878 TI - Role of calcium in angiotensin II-mediated aldosterone secretion. PMID- 2558879 TI - Effect of particle size on quartz-induced hemolysis and on lung inflammation and fibrosis. AB - To assess the role of crystal size in biologic responses, we quantitated red blood cell lysis and lung inflammation and fibrosis in the mouse using 4 alpha quartz preparations with average diameters of 1, 5, 7.8, and 11.2 microns. When compared on the basis of identical crystal surface areas, the 1-micron fraction was more hemolytic than the other 3 fractions. The three larger fractions had equivalent membranolytic activities. After 6 weeks of postintratracheal instillation of the crystals into mice, the 1-micron-diameter crystal fraction increased wet lung weights by 1.25 x that of saline controls, while a 1.75 x increase was found for the three larger crystal fractions. A similar response was found when evaluating fibrosis development by determining lung hydroxyproline levels. Measurement of the percentage of the crystal dose remaining in the lungs revealed that the biologic differences observed were not due to a difference in the clearance of the smaller crystal fraction. Thus, larger crystals of alpha quartz produce a greater degree of inflammation and fibrosis when instilled into the lung than those of 1 micron diameter, even though the smaller crystals are more membranolytic in vitro and appear to be cleared from the lung at the same rate as the larger crystals. PMID- 2558880 TI - Functional and morphologic comparison of silica- and elastase-induced airflow obstruction. AB - We have previously shown that intratracheally instilled silica (quartz) produces both morphologic evidence of emphysema and small-airway changes, and functional evidence of airflow obstruction. To further define the nature of the airflow associated lesions induced by silica, we compared silica-exposed rats to rats given intratracheal elastase, a standard model of emphysema. Both silica and elastase produced increases in RV and FRC along with upward shifts in the pressure-volume curves. Flows were decreased in both groups, but the changes were more severe in the silica-treated animals. Morphologically, the two treatments produced about the same degree of airspace enlargement, present in both alveoli and alveolar ducts, as well as decreased alveolar and alveolar duct surface area/unit lung volume. Elastic fiber length per unit volume was also decreased in both groups. However, small-airway walls were markedly thickened in the silica treated compared to the elastase-treated group. We conclude that, in the parenchyma, both these agents produce morphologically similar airspace dilatation, and both induce destruction of elastic fibers. These lesions appear to correlate with abnormalities in the pressure-volume curve. Flow reductions seem to be primarily correlated with changes in small-airway wall structure. PMID- 2558881 TI - Postnatal changes in an alpha subunit isoform, alpha(S), of Na+,K(+)-ATPase in the submandibular gland of rats. AB - The alpha and alpha(+) isoforms of Na+,K(+)-ATPase were isolated from the kidney and brain of rats and purified. Their antisera were raised to analyze the alpha isoforms in rat tissues. We found that the submandibular gland (SMG) contains a new immunoreactive alpha subunit isoform, designated alpha(S) in this report, in addition to alpha identical with those found in the kidney or brain. The new alpha(S) strongly reacted with anti-alpha-antiserum but to a much lesser extent with anti-alpha(+)-antiserum. The alpha(S) had a slightly lower molecular weight (approximately 90,000) than the brain and kidney alpha isoforms. Various fractions of SMG tissues were added to the SMG microsomes and incubated in order to test whether or not the alpha(S) is formed artificially; no increase of alpha(S) was observed by these treatments, suggesting that the alpha(S) was not the product formed from alpha during the preparation of microsome sample, but was rather a protein originally present in the SMG. The alpha(S) protein was not detected in the SMG of 2- or 5-week-old rats, but it gradually increased in rats older than 8 weeks, reaching the maximum in 30-week-old animals. The Na+,K(+) ATPase activity in the SMG increased concomitantly with the increase of alpha(S), indicating that Na+,K(+)-ATPase comprising alpha(S) also shows enzyme activity; it is speculated that alpha(S) may have some unique and unknown function(s) in older rats. PMID- 2558882 TI - Dietary salt and the glycaemic response to meals of different fibre content. AB - In attempting to resolve the existing controversy on the effect of dietary salt intake on glycaemic responses, we investigated post-prandial plasma glucose levels in 10 healthy normal weight non-diabetic Nigerian subjects (aged 23.1 years +/- 1.3 (s.e.m.) with body mass index, BMI 19.9 +/- 0.6 kg/m2) consuming equal amounts of carbohydrate from glucose, boiled yam (low fibre content of 0.9 per cent raw tuber weight) and boiled black-eyed peas (high fibre content of 4.8 per cent dry weight) with and without added table salt (4.25 g). The results indicated no significant differences in fasting, peak and 2-h plasma glucose concentrations and total and incremental areas under the 3-h glucose/time curves in the subjects consuming each meal with and without added salt. Added salt had no influence on the glycaemic index of each meal. We conclude that salt has no effect on the glycaemic response to plain glucose or meals with varying fibre content even in a population known to demonstrate defects in salt handling. PMID- 2558884 TI - Polyacrylamide gradient gel electrophoresis of cytosolic retinol- and retinoic acid-binding proteins: application to rat testis and liver. AB - Distribution and cellular levels of retinol-binding protein and retinoic acid binding protein, involved in the molecular action of retinoids, were analyzed in rat testis and liver. Both binding proteins of cytosolic extracts were separated by linear-polyacrylamide gradient gel electrophoresis and following electrophoretic separation, could be visualized by complementary identification tests such as autoradiography and marker proteins. The concentration of the binding proteins were evaluated by scanning the polyacrylamide gradient gels and the resulting data were found to be in accordance with those obtained by counting radioactivities. Polyacrylamide gradient gel electrophoresis appears suitable to detect and quantitatively evaluate cytosolic retinol- and retinoic acid-binding proteins. PMID- 2558883 TI - A procedure for analysis of densitometric spectra. AB - Computer-aided quantitative analysis of densitometric spectra is presented. The densitometric spectra are decomposed into component bands using the Powell and Marquardt minimizers. Several different functions for the component bands are utilized. It is shown that the densitometric spectra can be decomposed into component bands with high accuracy only if the proper shapes of the bands are chosen. The method described was used for quantitative analysis of densitometric spectra of DNA cleaved by neocarzinostatin. The procedure is general and can be applied to analyses of autoradiographic films and to direct scans of electrophoretic gels. It is shown that densitometric spectra which show highly overlapped bands are well approximated by asymmetric Cauchy and Gauss functions. Well separated densitometric bands which have substantial asymmetry can be fitted to more sophisticated shapes formed by combinations of symmetric and asymmetric Cauchy and Gauss functions. High accuracy fitting to asymmetric densitometric curves may only be achieved using the cosine function introduced by Mignot and Rondot, J. Appl. Cryst. 1976, 9,460-465. PMID- 2558885 TI - Analysis of virus protein heterogeneity among group B coxsackie viruses using a "mini" two-dimensional gel electrophoresis system. AB - The application of a small format two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) system to the study of protein heterogeneity among group B coxsackie virus (CVB) isolates is described. Under the conditions of electrophoresis developed during this study, protein samples could be processed within 7 h and up to 300 intracellular proteins were resolved from uninfected HEp 2 cell lysates. 2D-PAGE was used to characterise the intracellular proteins of clinical CVB isolates of serotypes 4 and 5. Intracellular proteins from virus infected cells were radiolabelled using a pulse-chase protocol under conditions which promoted inhibition of cellular protein synthesis. Depending on the CVB serotype up to 11 intracellular virus proteins were identified, ranging in molecular weight between 14,000 and 54,000. Although the overall two-dimensional protein profiles were characteristic for the two CVB serotypes, within a CVB serotype there was some heterogeneity of the virus proteins, mainly affecting the proteins' net charge. The sensitivity of 2D-PAGE in detecting subtle differences in virus proteins combined with the convenience of the small gel format makes this a suitable approach for the study of the molecular epidemiology of human virus pathogens. PMID- 2558886 TI - Transferrin receptors and cation-independent mannose-6-phosphate receptors deliver their ligands to two distinct subpopulations of multivesicular endosomes. AB - The distribution of transferrin receptors (Tf-R) was determined in Clone 9 hepatocytes and compared to that of 215 kDa, cation-independent mannose-6 phosphate receptors (M6P-R) by double labeling. Cells were allowed to take up exogenous human transferrin (Tf) for 5 to 30 min, after which Tf, Tf-R, and M6P-R were localized by immunofluorescence using specific antibodies. All these proteins were found to be concentrated in the juxtanuclear or Golgi region. When Clone 9 cells were treated with NH4Cl to trap M6P-R in endosomes (Brown, W. J., J. Goodhouse, M. G. Farquhar: J. Cell Biol. 103, 1235-1247 (1986)), the distribution of the two receptors differed: Tf-R remained the same as in controls, but M6P-R were localized in large vacuolated endosomes. To carry out double labeling experiments at the electron microscope level, transferrin gold conjugates (Tf-Au) were prepared, and M6P-R were detected by immunoperoxidase labeling. Tf-Au binding to the cell surface was specific as it was reduced approximately 70 to 79% in the presence of excess native Tf. When Clone 9 cells were incubated with Tf-Au at 37 degrees C for 5 to 30 min, or binding of Tf-Au was carried out at 4 degrees C followed by warming to 37 degrees C, Tf-Au was found within a peripheral tubulovesicular network and within multivesicular endosomes that were not labeled with anti-M6P-R. Other multivesicular endosomes of similar size and morphology were heavily labeled for M6P-R but contained little or no Tf-Au. Tf-Au and M6P-R were also found in separate endosomes in cells treated with NH4Cl. Native Tf was localized in the same compartments as Tf Au by immunoperoxidase labeling of both Clone 9 cells and mouse myeloma cells. We conclude that in Clone 9 hepatocytes, Tf/Tf-R internalized from the cell surface and M6P-R bearing newly synthesized lysosomal enzymes from the Golgi deliver their ligands to two different subpopulations of multivesicular endosomes. The endosomal subpopulation visited by Tf/Tf-R is known to correspond kinetically to early endosomes. The endosomal subpopulation heavily labeled for M6P-R presumably represent a later endosomal compartment which serves as the junction point where endocytosed ligands and newly synthesized lysosomal enzymes enroute to lysosomes meet. PMID- 2558887 TI - Cytochemical study of the localization and organization of parental herpes simplex virus type I DNA during initial infection of the cell. AB - Changes in the location and structural organization of parental herpes simplex virus type 1 (HSV-1) DNA during its migration from the extracellular space to the interior of the nucleus of the target cell were examined by in situ hybridization using an HSV-1 DNA probe, specific DNA staining, and autoradiography after infection of cells with tritium-labeled viruses. In situ hybridization was carried out on denatured DNA to reveal as much as possible of the HSV-1 sequence present at the surface of the sections, and also on non-denatured DNA which revealed the presence of single-stranded portions of parental DNA, both prior to and during its intracellular migration. The results from in situ hybridization and autoradiography demonstrated that a short interval of about 15 min separated the initial contact of the viruses with the cells from the entry of parental viral DNA into the nucleus. In transit, morphologically intact nucleoids were released into the cytoplasm, and swollen nucleoids which contained partially decondensed viral DNA became detectable in the juxtanuclear cytoplasm and the periphery of the nucleus among the cell chromatin fibers. Completely decondensed parental viral DNA fibers could not be distinguished structurally from cellular DNA, but their position could be revealed by the in situ hybridization label. The infective DNA became randomly distributed within all compartments of the nucleus except the matrix-associated clusters of interchromatin granules. PMID- 2558889 TI - Terminal elimination plasma half-life of delta 1-tetrahydrocannabinol (delta 1 THC) in heavy users of marijuana. AB - The terminal elimination half-life of delta 1-tetrahydrocannabinol (delta 1-THC) was investigated in eight men who were heavy users of marijuana. A stable isotope assay, following smoking deuterium-labeled delta 1-THC, was used to determine plasma concentrations. In two additional users plasma levels were followed after administration of unlabeled delta 1-THC. The subjects were asked to smoke a "loading dose" of 56 mg delta 1-THC during two days and then abstain from all marijuana use for 4 weeks. The pharmacokinetic behavior was consistent with a multicompartment model with a mean plasma elimination half-life of delta 1-THC of 4.3 days when concentrations were followed for 10-15 days after smoking. In the two subjects with detectable plasma levels during 4 weeks, half-lives of 9.6 and 12.6 days was obtained. PMID- 2558888 TI - Wilms tumour in a patient with growth hormone replacement therapy. AB - Wilms tumour was found in a Japanese boy aged 5 years 9 months with isolated growth hormone (GH) deficiency and some congenital anomalies. He had received pituitary GH replacement therapy from the age of 2 years 1 month to 4 years 7 months and after a 1 year interval he received biosynthetic GH for 2 months until the tumour became clinically apparent. This was the sixth known patient with GH deficiency to develop a malignant neoplasm during or after GH replacement therapy and the first with a solid tumour in Japan since 1975, when treatment with pituitary GH for patients with GH deficiency was introduced. PMID- 2558890 TI - Lack of effect of repirinast on the pharmacokinetics of theophylline in asthmatic patients. AB - A possible pharmacokinetic interaction between theophylline and repirinast has been investigated in asthmatic patients. The kinetics of theophylline was studied in seven adult in-patients given theophylline 400-800 mg b.d. alone and after three weeks of co-administration of repirinast. There was no effect on the kinetics of the combined treatment. PMID- 2558891 TI - Rapid development of brain beta-adrenoceptor down-regulation induced by phenelzine: subcellular studies. AB - The distribution of the specific binding of [3H]dihydroalprenolol in sucrose gradients containing homogenates of rat cerebral cortex centrifuged to equilibrium was examined after acute treatment with phenelzine (50 mg/kg i.p.). There was a time-dependent decrease in the number of beta-adrenoceptors in the plasma membrane fractions (0.8-1.0 M sucrose), without any significant alteration in the light density fractions (0.4-0.5 M sucrose). These results suggest that agonist-induced beta-adrenoceptor down-regulation is promoted by a process other than receptor internalization. PMID- 2558892 TI - Solubilization of endothelin/sarafotoxin receptors in an active binding form. PMID- 2558893 TI - Glutamine mimics glycine to enhance [3H]TCP binding at the NMDA receptor complex. PMID- 2558894 TI - Clonidine-induced hypoactivity and mydriasis in mice are respectively mediated via pre- and postsynaptic alpha 2-adrenoceptors in the brain. AB - Since brain alpha 2-adrenoceptors occur both pre- and postsynaptically, experiments were carried out to determine the synaptic locations of those receptors mediating clonidine-induced hypoactivity and mydriasis. Intraperitoneal (i.p.) injection of clonidine (1-3000 micrograms/kg) to mice dose dependently induced these two responses and also decreased brain concentrations of 3-methoxy 4-hydroxyphenylglycol (MHPG). The ED50 values were: 120 micrograms/kg for hypoactivity (95% confidence limits 103-140 micrograms/kg), 54 micrograms/kg for mydriasis (95% confidence limits 40-74 micrograms/kg) and 18 micrograms/kg for MHPG reduction (95% confidence limits 8-36 micrograms/kg) suggesting that these responses could all be presynaptically mediated. However, methamphetamine which increases noradrenaline turnover was found to dose dependently produce mydriasis, but not hypoactivity, after peripheral (0.1-5 mg/kg i.p.) or central (0.5-10 micrograms i.c.v.) injection. The mydriasis produced by methamphetamine (0.5 mg/kg i.p.) was abolished by i.c.v. injection of 1 micrograms idazoxan or yohimbine, but not 2.5 micrograms prazosin or pindolol, showing this effect was mediated by central alpha 2-adrenoceptors. Methamphetamine (1-10 micrograms i.c.v.) potentiated the mydriasis induced by clonidine (50 micrograms/kg i.p.) suggesting this was a postsynaptic alpha 2-adrenoceptor response. By contrast, methamphetamine (1-10 micrograms i.c.v.) dose dependently reversed clonidine (100 micrograms/kg i.p.) hypoactivity indicating this response was mediated by presynaptic alpha 2-adrenoceptors. These hypotheses were confirmed by destruction of noradrenergic neurones using DSP-4 (100 mg/kg i.p. x 2). This treatment prevented the mydriasis response to methamphetamine (0.5 mg/kg i.p.), but not clonidine (100 micrograms/kg i.p.) and markedly attenuated clonidine (100 micrograms/kg i.p.) hypoactivity. PMID- 2558895 TI - Effects of a selective kappa-opioid agonist, U-50,488H, on morphine dependence in rats. AB - Rats were made dependent on morphine by mixing the drug with their only source of food. Naltrexone (0.5 mg/kg) injection precipitated a syndrome of withdrawal signs including weight loss. Pretreatment with the selective kappa agonist, U 50,488H (1.0, 30.0 or 10.0 mg/kg), generally had no effects on the signs of morphine withdrawal. In other subjects, U-50,488H was repeatedly administered (1.0, 3.0 or 10.0 mg/kg per 12 h) during the development of morphine dependence. In these subjects, the course of naltrexone-precipitated withdrawal was unchanged. These results suggest that agonist activity at kappa receptors is not sufficient to alter morphine dependence or withdrawal. PMID- 2558896 TI - Peripheral benzodiazepine binding sites in Nb 2 node lymphoma cells: effects on prolactin-stimulated proliferation and ornithine decarboxylase activity. AB - [3H]Ro 5-4864 binds to Nb 2 node lymphoma cells in a specific saturable and reversible fashion. Scatchard analysis of specific binding data reveals a single, homogeneous class of whole cell binding sites with a Kd of 3.94 +/- 0.22 nM and a Bmax value of 155 +/- 11 fmol (Ro 5-4864 bound)/2 x 10(6) cells. Ro 5-4864, a reported peripheral benzodiazepine receptor agonist both inhibits (10(-6) M) and potentiates (10(-9) M) the mitogenic action of prolactin on the Nb 2 node lymphoma cells. Interestingly, PK 11195, an antagonist, potentiates (10(-9) M) the mitogenic activity of prolactin in these cells. The actions of both Ro 5-4864 and PK 11195 seem to be mediated through a common receptor type since a 10(-6) M concentration of either agent will block the others potentiating action. Furthermore, the simultaneous addition of a 10(-9) M concentration of Ro 5-4864 and PK 11195 does not further increase the effect on prolactin stimulated mitogenesis. Clonazepam, a central benzodiazepine receptor agonist has no effect on prolactin-stimulated mitogenesis in this system. These data suggest that the Nb 2 node lymphoma cells possess a peripheral-type benzodiazepine receptor. In these cells, this receptor seems to serve the function of modulating the ability of the growth factor, prolactin to initiate the mitogenic process. These studies also suggest that Ro 5-4864 is functioning as a partial agonist rather than a 'pure' agonist for the peripheral benzodiazepine receptor in this system. PMID- 2558897 TI - Biologically active conformation of [Met5]- and [Leu5]enkephalin on delta opioid receptor. AB - Investigations of the conformation of endogenous enkephalins are generally based on structural comparisons of enkephalins with other opiates and on experimental pharmacological studies. Based on such investigations, we now propose a novel model of the biologically active (receptor-bound) conformation of [Met5]- and [Leu5]enkephalin on the delta opioid receptor. The model helps with the design of new opioid analgesics. PMID- 2558898 TI - Pertussis toxin treatment counteracts intramembrane interactions between neuropeptide Y receptors and alpha 2-adrenoceptors. AB - The effect of intracerebroventricular injections of pertussis toxin were investigated on the neuropeptide Y-induced modulation of alpha 2-adrenoceptor binding in membranes from the dorsomedial medulla oblongata of the rat. Concentration-response experiments showed that neuropeptide Y reduced the binding affinity of the alpha 2-agonist, p-[3H]aminoclonidine, with a maximal effect of 30% at 3-30 nM. Pertussis toxin treatment (10 micrograms, 24 h) counteracted this modulation, without reducing the binding of neuropeptide Y to its own receptor. The results indicate that pertussis toxin-sensitive G-proteins are essential for the mediation of the intramembrane interaction between neuropeptide Y receptors and alpha 2-adrenoceptors. PMID- 2558899 TI - 4-Aminopyridine induces the release of neuropeptide Y (NPY) to produce an atropine- and tetrodotoxin-resistant contraction in rabbit isolated jejunum. AB - 4-Aminopyridine (4-AP) induced an atropine- and tetrodotoxin (TTX)-insensitive contraction (resistant contraction), in a concentration-dependent manner, in the isolated jejunum of rabbits. The failure of specific antagonists of histamine, serotonin and substance P to affect this resistant contraction ruled out the participation of histamine, serotonin and/or substance P. Antiserum against neuropeptide Y (NPY) reduced this resistant contraction in a concentration dependent manner and inhibited the action of 4-AP totally at a high concentration (1.25% dilution) whereas normal serum lacked this ability. This suggested that the release of NPY was involved in this 4-AP-induced resistant contraction. Radioimmunoassay of NPY-like immunoreactivity in isolated synaptosomal preparations indicated that 4-AP possessed the ability to induce the release of NPY. However, guanethidine did not affect the actions of 4-AP, indicating that NPY is released mainly from non-adrenergic nerves. Our results indicate that 4-AP induces the release of NPY from non-adrenergic nerves to produce an atropine- and TTX-resistant contraction in the isolated jejunum of rabbits. PMID- 2558900 TI - Isolated human bladder: evidence for an adenine dinucleotide acting on P2X purinoceptors and for purinergic transmission. AB - In isolated strips of human urinary bladder detrusor muscle, ATP, alpha, beta methylene ATP and P1,P6-diadenosine hexaphosphate caused concentration-dependent contractions. ATP was less potent than the two synthetic purine compounds and gave smaller maximum responses. Responses to ATP, P1,P6-diadenosine hexaphosphate and noncholinergic nerve stimulation were blocked following desensitization of P2X-purinoceptors by alpha,beta-methylene ATP. Thus, adenine dinucleotides can act on P2X-purinoceptors and there is an element of purinergic neuromuscular transmission in the human urinary bladder. PMID- 2558901 TI - Modulation of uterine GABAA receptors during gestation and by tetrahydroprogesterone. AB - Binding of a GABAA receptor agonist, [3H]muscimol, was studied in rat uterine membranes of non-pregnant rats and those at days 15 and 19 of pregnancy. Also, an interaction of the uterine GABAA receptors with tetrahydroprogesterone was examined. At day 15 of gestation [3H]muscimol binding was twice as high as in non pregnant rats, but at day 19 it was reduced. These changes resulted from an increase of the receptor affinity and decrease of the receptor density, at days 15 and 19, respectively. Since tetrahydroprogesterone, in vitro, also increased [3H]muscimol binding, we propose that this steroid may participate in regulation of uterine function during pregnancy. PMID- 2558903 TI - Age-related changes in calpain II and calpastatin in rat lens. AB - The purpose of these experiments was to determine how the activity and regulation of calpain in rat lens changed during aging. Calpain II enzyme activity and immunoreactivity decreased with both chronological and anatomical age. Two pieces of data suggested that loss of soluble calpain II was a result of both autolysis and insolubilization during aging: (i) proteolytic fragments of calpain were detected in lenses with molecular weights similar to fragments produced during incubation of purified calpain II with calcium; (ii) the water-insoluble fraction of lens cortex contained increasing amounts of calpain antigen during aging both the 75-kDa calpain subunit and a unique high-molecular-weight immunoreactive protein. The regulation of calpain II also appeared to change with age. The activity of calpain II in vivo may be regulated by the relative concentrations of calpain II and its endogenous inhibitor calpastatin. Calpain II concentrations decreased in the rat lens with age, whereas levels of the endogenous inhibitor calpastatin were maintained. Assays of calpain II and calpastatin indicated that upon aging there was insufficient activity of calpain II to overcome the inhibition of calpastatin in the nucleus. These findings were confirmed by incubation of crude lens homogenates of 2-week- and 7-month-old rat lens regions with calcium. It is hypothesized that binding of calpain II to membrane may be important for calpain II activation, especially in older lens regions, because it may allow escape from the inhibitory action of calpastatin. PMID- 2558902 TI - Presynaptic alpha 2-adrenoceptor and kappa-opiate receptor occupancy promotes closure of neuronal (N-type) calcium channels. AB - Synaptosomes prepared from rat cerebral cortex by homogenization in isotonic sucrose and centrifugation on four-step discontinuous percoll density gradients were loaded with the fluorescent indicator fura-2 to allow measurement of intrasynaptosomal free calcium concentrations [( Ca2+]i). Incubation of fura-2 loaded synaptosomes with either the kappa-opiate agonist U-50,488H (0.1-100 microM) or the alpha 2-adrenoceptor agonist clonidine (0.1-100 microM), resulted in a dose-dependent reduction in [Ca2+]i and these changes were completely antagonised by prior inclusion of naloxone (20 microM) or idazoxan (RX781094) (2 microM) respectively. When the 1,4-dihydropyridine Ca2+-channel blocker nifedipine (1 microM) was incubated with synaptosomes for 1 min, there was a 17.0% decrease in [Ca2+]i and when it was combined with either U-50,488H (1 microM) or clonidine (1 microM) there was a reduction in [Ca2+]i of 35.0 and 48.1% respectively i.e. the effects were additive. The increases in the depression of [Ca2+]i produced by these drug combinations were antagonised by the inclusion of naloxone (20 microM) or idazoxan (2 microM) which resulted in decreases in free [Ca2+]i of 26.5 and 14.1% respectively. These data indicate that the effects of clonidine and U-50,488H are not mediated by L-type Ca2+ channels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558904 TI - Regional differences in the morphology and enzyme distribution of the spiny dogfish (Squalus acanthias) ciliary epithelium. AB - The ciliary epithelium of spiny dogfish eyes has previously been used for studies of epithelial electrolyte transport and the results are found to be comparable to those obtained in mammals. In this study we report the ultrastructure and enzyme histochemistry of the ciliary epithelium of spiny dogfish in comparison with that of mammals. In contrast to the mammals, the lens in spiny dogfish is connected to the anterior and middle part of the ciliary epithelium by a broad zonula-like suspensory ligament and by a short ligament to the ventral papilla, while the posterior region is mainly free of zonula-like fibers. The non-pigmented epithelium (NPE) of the posterior region shows numerous membrane infoldings, interdigitations and many mitochondria in the cytoplasm as well as histochemical staining for Na+ K(+)-ATPase and carbonic anhydrase (CA). These findings are similar to those seen in the anterior pars plicata of mammals which is mainly involved in aqueous secretion. In the anterior and middle part, the NPE cells have only a few membrane infoldings and few mitochondria in the cytoplasm, but abundant surfaces of rough endoplasmic reticulum, numerous ribosomes and Golgi material, indicating protein synthesis. Histochemically the cells stain for Na+, K(+)-ATPase but not for CA. These findings are comparable to the pars plana in mammals. In contrast to mammals, where the pigmented epithelium (PE) shows nearly no staining for Na+, K(+)-ATPase but an intensive one for CA, the PE cells of spiny dogfish are heavily stained for Na+, K(+)-ATPase in all regions of the eye, but show nearly no staining for CA. PMID- 2558905 TI - Human retinal capillary basement membrane leaflets are morphologically distinct: a correlated TEM and SEM analysis. AB - Because retinal capillaries and their associated basement membranes (BMs) are significantly altered in a number of diseases (most notably diabetic retinopathy), the human retinal microvasculature is of interest to both basic scientists and clinicians. Consequently, numerous TEM studies centered primarily on cellular elements of retinal microvessels have been carried out. Ultrastructural studies emphasizing retinal capillary extracellular matrix (ECM) materials including BMs however, are nearly non-existent. Accordingly, the current correlated TEM/SEM investigation was undertaken. The study shows that retinal capillary walls are comprised of a continuous layer of endothelial cells and a discontinuous layer of intramural pericytes which are in frequent contact. These are underlain and/or surrounded by a retinal capillary BM complex which includes pericytic matrix, fibrillar collagen, and subendothelial, pericytic and Muller cell BM leaflets. Following sequential detergent treatment, all retinal cells are solubilized. Vessel ECM components, however, maintain their in vivo histoarchitectural relationships. Moreover, on the basis of substructure, susceptibility to non-specific proteases and anionic site density, BM leaflets are morphologically distinct. In addition, high-resolution SEM studies show that BM surface characteristics are tissue specific. It is concluded that retinal capillary BM complexes are comprised of structurally unique subcomponents the characteristics of which should be considered in future studies of retinal capillary BM structure, composition and function and particularly in investigations in which retinal capillaries are pathologically altered. PMID- 2558906 TI - Non-allelism of three genes (rcd1, rcd2 and erd) for early-onset hereditary retinal degeneration. AB - Cross-breeding experiments were utilized to study the genetics of three autosomal recessive, early onset retinal degenerations in dogs. Irish setters affected with rod-cone dysplasia type 1 (rcd1) were bred to Norwegian elkhounds affected with early retinal degeneration (erd). All offspring (15 pups-two litters) surviving to diagnostic age were phenotypically normal, as assessed by electroretinography, retinal morphology and assay of retinal cyclic nucleotide content. One phenotypically normal female Irish setter-Norwegian elkhound crossbred dog (the progeny of the above breeding and thus heterozygous at both the rcd1 and the erd locus) was bred to a collie dog affected with rod-cone dysplasia type 2 (rcd2). All 11 pups from this breeding also proved phenotypically normal by the above methods. These results establish that the genes rcd1, rcd2 and erd are non allelic. Biochemical data are also presented that establish that erd, unlike rcd1 and rcd2, is not associated with abnormal metabolism of retinal cyclic guanosine monophosphate (cyclic GMP). PMID- 2558907 TI - Distribution of cholinergic receptors in the rat and human neocortex. AB - Autoradiographic labelling of muscarinic (M1, M2, NMS binding sites) and nicotinic receptors shows an inhomogeneous distribution over architectonically identified cortical areas of the rat brain with highest concentrations in the medial prefrontal and frontal areas. Beside this general trend the areal patterns of different receptors are slightly varying. The laminar distribution of these receptors in the rat and human neocortex is characterized by two different patterns, one with highest receptor densities in the supragranular layers (M1 receptors, NMS binding sites), the other with a preferential labelling of layer IV and (with a lower intensity) layer V (M2 and nicotinic receptors). M1 receptors and NMS binding sites are codistributed at the laminar level with each other and with GABAA, D1, 5-HT1 and glutamate receptors; M2 receptors are codistributed only with nicotinic receptors. Immuno-histochemical studies with antibodies against muscarinic and nicotinic receptors demonstrate that these structures occur mainly in pyramidal and spiny stellate cells and to a lesser extent (13%) in a variety of interneurons. The immunoreactivity is visible in the perikaryon, dendrites and postsynaptic membranes. Neurons are found in the human neocortex, which react exclusively with one of the two antibodies, but a fraction of the neurons (about 30%) contains antigenic sites reacting with both antibodies. This is interpreted as colocalization of nicotinic and muscarinic receptors in some cortical neurons. PMID- 2558908 TI - Topography of beta NGF receptor-positive and AChE-reactive neurons in the central nervous system. AB - Recent reports have led to widespread interest in the role of beta-nerve growth factor (beta NGF) in the central nervous system. To learn more about the action of beta NGF in the central nervous system we have mapped the distribution of beta NGF receptors and compared it with that of acetylcholinesterase (AChE), a sensitive enzyme marker for cholinergic neurons. In situ autoradiography revealed strong and saturable beta NGF binding to several groups of neurons in basal forebrain and brainstem. They also contain significant levels of mRNA coding for beta NGF receptors. beta NGF receptors and AChE are codistributed on the medial septal nuclei and in the basal forebrain, including the striatum. In the brainstem, beta NGF receptors are present on the neurons in the lower part of the reticular formation and in cochlear nuclei but do not correspond to the distribution of AChE reactivity. PMID- 2558909 TI - Mediation of acetylcholine's excitatory actions in central neurons. AB - In experiments on the hippocampus in situ (in rats under urethane), neither cyclic GMP nor H-8 (an antagonist of cyclic nucleotide-dependent kinases) had much effect on CA1/CA3 population spikes or on the excitatory action of ACh. This is further evidence against the idea that cyclic nucleotides play a major role as cholinergic second messengers. On the other hand, the results of tests with a PKC antagonist sphinganine are in keeping with some involvement of PKC in cholinergic actions. (Another PKC antagonist, H-7, proved to be a very powerful excitant, probably via disinhibition). Preliminary experiments on CA1 neurons in hippocampal slices (by single electrode voltage clamp), confirmed previous reports that carbachol depresses A- and C-type K currents, as well as inward Ca2+ currents; though the latter effect was sometimes mainly due to frequency dependent inactivation of Ca currents. It is suggested that a single, primary muscarinic action, the acceleration of phosphinositide turnover, may account for a variety of secondary effects: on the one hand, via activation of PKC, a number of possible PKC-mediated actions, such as block of the slow AHP; on the other, via IP3 formation, a block of IM and a rise in cycloplasmic free Ca2+ that may cause inactivation of both Ca2(+)-inward currents, and Ca2(+)-dependent GKs. PMID- 2558910 TI - Opposing effects of acetylcholine on the two classes of voltage-dependent calcium channels in hippocampal neurons. AB - Acetylcholine (Ach) was tested for its effect on calcium currents in primary cultures of embryonic rat hippocampal neurons. Ach reversibly depressed, in a dose-dependent way, the high voltage activated (HVA) Ca currents. The effect was antagonized by atropine. Our results suggest that a pertussis toxin (PTX) sensitive GTP binding protein (G-protein) is involved in the signal transduction mechanism between the Ach receptor and the HVA Ca channel. Activating rather than depressive effects of Ach were observed on the low voltage-activated component of Ca currents. This effect was also antagonized by atropine but is not mediated by a PTX-sensitive G-protein. PMID- 2558911 TI - [Current state of the problem of cortical inhibition]. AB - Results concerning neuronal, synaptic and ionic mechanism of inhibition in cortical neurons obtained during the last years are presented. PMID- 2558912 TI - [Reactions of cat motor cortex neurons to stimulation of the pyramidal tract and ventroposterolateral thalamic nuclei]. AB - Responses of neurons of motor cortex evoked by stimulations of pyramidal tract (PT) and ventroposterolateral (VPL) nucleus of thalamus were studied in cats immobilized by Myorelaxin. Antidromic spikes were found in 22.6% and in 9.9% of cortical cells when PT and VPL were stimulated, respectively. Fast- and slow conducting PT-neurones could be differentiated according to antidromic excitation latencies. PT stimulation evoked EPSPs in 46.3% of studied neurones and VPL stimulation--in 48.2% ones. Monosynaptic EPSPs were identified in responses of fast- and slow-conducting PT-units and of neurones projecting in VPL; mechanisms and functional role of such reactions are discussed. Di- and polysynaptic IPSPs were evoked in 74.5% of units by PT stimulation and in 94.4%--by VPL stimulation. Three groups of IPSPs were classified with durations to 120, 130-280 and more than 300 ms. Duration of PT-evoked IPSPs was higher in cortical neurones from surface layers and VPL-evoked ones--in units localized in deep layers. PMID- 2558913 TI - [Effect of damage of the nigro-neostriatal dopaminergic system by MPTP on the transmission of corticofugal impulses to the neurons of the caudate nucleus]. AB - It is shown in acute experiments on cats (males) that the induced responses as action potentials (AP) by the latent period (LP) less than 8.0 ms in the caudate nucleus neurons (CN) to a single stimulation of the motor zone of the cortex (MI) are more frequently inhibited than facilitated after specifying single stimulation of the compact part of the black substance (BS) in the intervals between stimuli 10-100 ms. As a result of system multiple injection of MPTP neurotoxin during 5 days per 5 mg/kg the number of CN neurons responding to stimulation of MI, AP, LP less than 8.0 Usec and to stimulation of BS-LP less than 10.0 ms reliably decreases. A conclusion is made that dopaminergic nigro striatum system exerts a protective action on the impulse transfer on monosynaptic connections from the cortex to striatum. PMID- 2558914 TI - [Reaction of neurons of the medial geniculate body to acoustic stimulation]. AB - Responses of medial geniculate body (MGB) neurons to pure tones and clicks were studied in acute experiments in immobilized cats, preliminary operations being performed under calypsol anaesthesia. MGB units were identified by their reactions to cortical zone AI and brachium of inferior colliculus stimulations. When tonal stimuli were applied relay neurons of pars principalis of MGB usually demonstrated either unimodal tuning curves with narrow frequency band or fragmental ones with several narrow bands. On-response with subsequent inhibition of the background activity or without such an inhibitory period was most frequent type of the reaction (66.6%) of relay MGB neurons to tonal stimulation. The group of relay neurons with the tonic type of reaction (9.1%) was classified for which the duration of tonic response depends on the duration of tonal stimulus. Change of the excitatory reaction to the inhibitory one when the characteristic tone frequency is changed by non-characteristic++ ones is supposed to be a mechanism supplying sharpness of tuning at relay MGB neurons. It is concluded that responses of acoustic cortical neurons to sound stimulation depend to a great extent on the pattern of impulsation that comes from MGB relay units. PMID- 2558915 TI - [The membrane potential of the secretory cells of the mamma under the action of mezaton and izadrin]. AB - The development of depolarization mesaton-induced and hyperpolarization isadrine induced changes of the membrane potential of the mice mammary gland secretory cells was followed by a decrease in transepithelial electrical resistance and in potassium ions activity in the alveoli. The isadrine-induced hyperpolarization could be reversibly depressed by a substitution of chloride ions with sulphuric acid ions. Neither mesaton nor isadrine altered the MP after blocking of the Na+/K+ pump. Catecholamines seem to exert a direct alpha-adrenomimetic and an indirect beta-adrenomimetic reversible blocking effect upon the activity of the Na+/K+-ionic pump. PMID- 2558916 TI - [The role of the posterior lateral thalamic nucleus in the rabbit in the mechanisms of information transmission in the visual cortex]. AB - In chronic alert rabbits, both transient and sustained influence of the nucleus lateralis posterior upon visual cortical responses to the light flashes, were shown. The transient influences were revealed by an increase of the responses amplitude in the nucleus lateralis posterior and by a decrease of the amplitude during the K depression. The sustained influences took part in forming of thalamo cortical responses in visual cortex. This reciprocity was particularly obvious during pulse stimulation of the nucleus lateralis posterior. The sustained influences were more significant than the transient ones. PMID- 2558919 TI - [Stimulation of sodium transport by cobalt ions in the skin of amphibia against a background of aldosterone action]. AB - Sodium transport across toad Bufo marinus and frog Rana temporaria skin was enhanced by aldosterone added to Ringer solution at the basal plasma membrane. Cobalt ions applied to the outer surface of amphibian skin have increased sodium transport stimulated by aldosterone. The regulatory sites of the sodium channel macromolecule seem to be located on both surfaces of the channel apical membrane. PMID- 2558918 TI - [Changes in the transsinusoidal fluid transfer in the cat liver during noradrenaline infusion]. AB - The blood microcirculation and the lymph flow were studied in the cat hemodynamically isolated liver perfused under conditions of a regulated venous outflow and i.a. and intraportal administration of noradrenaline. An increase in the fluid transsinusoidal transfer revealed by the lymph flow acceleration in the liver, was determined by an increased hydrostatic pressure in the sinusoids at practically unaltered level of sinusoidal filtration. The above shifts were observed against the background of constriction of arterial, portal, venous and lymphatic vessels and of a decrease in capacitance of the liver vessels which were due to alpha-adrenocereptors excitation by noradrenaline. PMID- 2558920 TI - The activity of some phosphatases in tissues of adult Hymenolepis nana Siebold (Csetoda). AB - Histochemical methods were used to study the localization and activity of acid and alkaline phosphatases, ATP-ase, 5-nucleotidase, and glucose-6-phosphatase in tissues of the mature form of Hymenolepis nana. Considerable differences in activity and localization of particular enzymes were observed in the organs of the parasite. The results obtained permit the statement that the integument is the most active enzymatically; in connection with the literature data, this gives grounds for the thesis that the integument of the cestodes functions as an absorbent-digestive organ. PMID- 2558917 TI - [The role of catecholaminergic synapses in the mechanisms of the formation of adaptation with the participation of polyphenol adaptogens]. AB - The analysis of the role of catecholaminergic synapses in the process of adaptation showed that the main point of the influence of polyphenolic adaptogens was a catechol-O-methyltransferase. Inhibiting the latter, the polyphenolic adaptogens exert a correcting effect on catecholaminergic (mainly dopaminergic) synapses and prevent the transmitter from reduction in them. PMID- 2558921 TI - Recurrent/persistent pneumonia in a 3 1/2-year-old-girl due to acquired immune deficiency syndrome. PMID- 2558922 TI - Diadenosine 5",5"'P1,P4-tetraphosphatase in Drosophila embryos: developmental regulation and characterization. AB - 1. An enzyme has been isolated from Drosophila embryos which specifically hydrolyzes dinucleoside tetraphosphates to the corresponding nucleoside tri- and tetraphosphates, with Km values around 4 microM. 2. Nucleoside mono-, di- and triphosphates are competitive inhibitors with K1 values i the 0.01 mM range. 3. The inhibition is particularly strong by adenosine tetraphosphate (Ki = 10 nM). 4. The enzyme is maximally active at pH 7.5 and is quite stable at acid pH. 5. The enzyme requires divalent cations for activity: Co(2+) much greater than [corrected] Mn(2+) Mg(2+) x Co(2+) stimulated about 90-fold at 6 mM. 6. The specific stimulation by Co(2+) has been described before, but at lower concentrations, for the enzyme of procaryotes which splits diadenosine tetraphosphate symmetrically. Zn(2+) and Ca(2+) are inhibitors of the Drosophila enzyme. Co(2+) is also inhibitor in the presence of Mg(2+). 7. The Drosophila enzyme has essential sulphydryl group(s) and a molecular weight of 26,000. 8. Diadenosine tetraphosphatase is present in mature oocytes and increases after fertilization to reach a peak 1.5 hr later. 9. From this time to 3.5 hr the activity decreased to remain at a plateau until the end of embryogenesis. 10. The profile of activity is compatible with its involvement in the regulation of nuclear division. PMID- 2558923 TI - Effects of epinephrine on glucose-1,6-bisphosphate and carbohydrate metabolism in skin. AB - 1. Injection of epinephrine induced in skin a decrease in the level of glucose 1,6-bisphosphate (Glc-1,6-P2), which was accompanied by correlated changes in the activities of several enzymes which are modulated by this regulator. 2. These effects were blocked by the alpha adrenergic blocker phentolamine, in contrast to muscle where the hormone increases Glc-1,6-P2, acting through beta receptors. 3. The changes in the enzymes' activities, as well as in glycogen and lactate content induced by epinephrine, reveal that the hormone causes, in skin, a stimulation of glycogenolysis and glycolysis, as well as an acceleration of pentose phosphate pathway. 4. The reduction in glycogen content induced by epinephrine, was blocked by the beta adrenergic blocker propranolol, whereas the hormone's effects on the other processes were mainly mediated through alpha receptors. PMID- 2558924 TI - Interaction of pyridoxal phosphate modified cytochromes c with mitoplasts. AB - 1. The stability of the native conformation of the heme crevice of pyridoxal phosphate (PLP)-ferricytochromes c as assayed by the pK, for 695 nm absorption band varies considerably. The pKa values are 8.76 for cytochrome c modified by PLP at lysine 79[PLP(Lys 79)-cyt. c], 9.23 for cytochrome c modified by PLP at lysine 86 [PLP(Lys 86)-cyt.c], 9.34 for doubly PLP substituted cytochrome c at lysines 79 and 86 [(PLP)2-cyt. c], 9.50 for triply substituted cytochrome c [(PLP)3-cyt. c] and 9.06 for native cytochrome c, which indicates less stable heme crevice of PLP-cytochrome c. 2. The singly PLP-modified cytochrome c indicate decreased activities with mitochondrial cytochrome c oxidase in the following order: PLP(Lys 86)-cyt. c less than PLP(Lys 79)-cyt. c less than native cytochrome c. The high affinity Km for PLP(Lys 86)-cyt. c, PLP(Lys 79)-cyt. c and native cytochrome c are 0.28 microM, 0.16 microM and 0.02 microM respectively. 3. PLP-cytochromes c show decreased binding affinities to fluorescence probes 12-(9 antroyl)-stearic acid and pyrene-labelled mitoplasts. The quenching of singly PLP modified cytochrome c depends significantly on the ionic strength. PMID- 2558925 TI - Effect of enzymatic methylation of apocytochrome c on holocytochrome c formation and proteolysis. AB - 1. Methylation of the lysine at residue 72 of yeast apocytochrome c increases its import into mitochondria. 2. Using methylated and unmethylated apocytochrome c as substrate and intact yeast mitochondria and a solubilized mitochondrial fraction as a source of cytochrome c heme lyase, the results show that the methylation state of the apoprotein has no significant effect on its conversion to holoprotein. 3. The above result suggests that the import mechanism is separate from the heme-attaching activity. 4. Unmethylated apocytochrome c was less resistant to a yeast homogenate fraction that methylated apocytochrome c, suggesting that methylation of apocytochrome c alters the conformation of the whole protein. PMID- 2558926 TI - Induction of c-fos-like protein in the rat adrenal cortex by acute stress- immunocytochemical evidence. AB - The effect of immobilization stress on the expression of c-fos protein in the adrenal cortex of adult rats was investigated immunocytochemically. After immobilization stress lasting for longer than 30 min, an enhanced c-fos-like immunoreactivity was observed in the cortical cells of the zona fasciculata and zona reticulata. Compared to unstressed controls, an about 5-fold increase in the density of the immunoreactive cells in a unit of the cortical area was seen following a 1-h immobilization. The enhanced immunoreactivity lasted for at least 3 h after 1-h immobilization and it began to diminish 5 h after the stress. Furthermore, administration of dexamethasone 2 h prior to 1-h immobilization attenuated the stress-enhanced immunostaining for the c-fos-like protein. These results suggest that an acute stress may cause a dramatic and long-persisting induction of c-fos-like protein in the cortical cells of rat adrenals. The characteristic zonal distribution of the c-fos induction in rat adrenals as well as the effect of dexamethasone suggest involvement of the pituitary adrenocorticotropic hormone (ACTH) in the induction. PMID- 2558927 TI - Reversal of the antagonism of deglycosylated human chorionic gonadotropin by aggregation with wheat germ agglutinin. AB - Human chorionic gonadotropin (hCG) behaves as an antagonist upon chemical deglycosylation with hydrogen fluoride. In this study it was found that the alpha and beta-subunits of deglycosylated hCG (DGhCG) still retained their ability to bind to wheat germ agglutinin (WGA) but not to concanavalin A. The antagonism of DGhCG against activation of adenylate cyclase and steroidogenesis was reversed, when WGA was bound to the N-linked carbohydrate moieties of hCG alpha- and beta subunits followed by addition of purified Leydig cells. The complex formed by incubation at an approximately equimolar ratio induced the maximal reversal of antagonism. This reversal of antagonism was diminished by addition of N acetylglucosamine. No increase of steric hindrance at the receptor sites was seen in binding studies of the [125I]DGhCG-WGA complex, indicating that the receptor binding domain of hCG may not be adjacent to the carbohydrate moieties. Kinetic studies of the hormonal response showed that the DGhCG-WGA complex terminated cAMP accumulation after 30 min of incubation, but not testosterone production. Our results suggest that tetravalent WGA can also reverse the antagonism of DGhCG, as in bivalent antibodies to hCG beta. PMID- 2558928 TI - The preparation of D-myo-inositol 1,4-bisphosphate and D-myo-inositol 1,4,5 trisphosphate in milligram quantities from a readily available starting material. AB - The optimisation of a reaction for the conversion of glycerophosphoinositols to phosphoinositols is described. This reaction has been used in a scheme, described in detail, for the formation of D-myo-inositol 1,4-bisphosphate and D-myo inositol 1,4,5-trisphosphate in mg quantities from a readily available preparation of mixed phosphoinositides. An optimised procedure is also detailed for the recovery of these products to high yield and purity. The identity of the products has been confirmed both by high resolution anion-exchange column chromatography and by 1H nuclear magnetic resonance studies. We report for the first time the 1H nuclear magnetic resonance spectrum for D-myo-inositol 1,4 bisphosphate. PMID- 2558929 TI - The antiglucocorticoid RU38486 is a potent accelerator of adipose conversion of 3T3-F442A cells. AB - We examined the effects of RU38486, a potent glucocorticoid and progestin antagonist, upon several aspects of 3T3-F442A adipocyte differentiation. RU38486 accelerated the onset of differentiation, as monitored by cell morphological changes, accumulation of lipid droplets and widespread increases in the rate of expression of several enzyme adipose markers and specific mRNAs. RU38486, at a maximal concentration of 1 microM, dramatically hastened the emergence of both fatty-acid synthetase (FAS) and glycerol-3-phosphate dehydrogenase (G3PDH) enzyme activities (550% and 450% above control values 4 days after confluence, respectively). RU38486 induction of G3PDH-specific activity ran parallel to an increase in G3PDH mRNA content (2.4-fold the control content 4 days after confluence). Moreover, RU38486-treated cells exhibited enhancement of adenylate cyclase sensitivity to both isoproterenol and ACTH (160% and 350% above control activities 8 days after confluence, respectively). While the level of expression of lipogenic markers reached similar values at the mature stage, RU38486 enabled cells to acquire hypersensitivity in terms of ACTH-stimulated adenylate cyclase activity. Similarly, adipsin gene expression was highly potentiated by the drug at day 15 post-confluence (5-fold the control value). RU38486 responsiveness observed in differentiating 3T3-F442A cells is dependent upon their prior developmental activation; none of the studied markers could be induced by the drug in the undifferentiating 3T3-C2 cell subclone. Finally, this antiglucocorticoid appears to be a useful tool for studies on adipose conversion in vitro; it could permit a re-evaluation of the role of glucocorticoids in the understanding of adipocyte development. PMID- 2558930 TI - Parathyroid hormone acute vascular effect is mediated by decreased Ca2+ uptake and enhanced cAMP level. AB - Cardiovascular effects of parathyroid hormone (PTH) have been recently described. Pharmacological doses of PTH both reduce arterial pressure and increase blood flow of vascular beds. Two possible cellular mechanisms were investigated: (a) transmembrane Ca2+ fluxes and (b) cyclic AMP response in vascular smooth muscle. In vivo, results in the rat show that injection of synthetic bovine 1-34 fragment of PTH (bPTH-(1-34] produced a rapid (1-2 min) but transient (5-16 min) hypotensive effect which was dose-related (0.4-4 nmol.kg-1). In the in vitro studies on isolated rat aorta, bPTH-(1-34) partially inhibited noradrenaline (NA) induced contractions by decreasing the sustained tonic component dependent on extracellular Ca2+. bPTH-(1-34) also produced relaxation of aorta preconstricted with NA or prostaglandin F2 alpha. Measurements of the lanthanum-resistant Ca2+ pool using 45Ca2+ showed that bPTH-(1-34) decreased basal Ca2+ uptake and partially inhibited Ca2+ uptake stimulated by NA or K+-depolarizing solution in a concentration-dependent fashion. In addition, bPTH-(1-34) caused a concentration related increase in cyclic AMP in rat isolated aortic tissues. Hypotensive and vasorelaxing effects of bPTH-(1-34) thus appear to be mediated by a decrease in the amount of Ca2+ available for contraction and by an increase in cyclic AMP response in vascular smooth muscle cells. PMID- 2558932 TI - Proglucagon expression, posttranslational processing and secretion in SV40 transformed islet cells. AB - HIT T15 is a B cell line derived from SV40 transformation of hamster islets. We describe here a HIT T15 variant, designated HIT T15-G, which appears to have evolved spontaneously and which expresses glucagon. Regulation of glucagon gene expression, posttranslational processing of proglucagon, and secretion of glucagon were studied in this cell line. Glucagon mRNA concentrations were increased approx. 2-fold following incubation of cells for 18 h in 10 microM forskolin but were unaffected by treatment with a phorbol ester (12-O tetradecanoylphorbol 13-acetate; TPA) or with ionomycin. Proglucagon was processed to glucagon, and several large molecular weight forms of GLP-I and GLP II which may include the major proglucagon fragment (MPF). The secretion of glucagon was stimulated by forskolin (5-fold), adrenalin (2-fold), arginine (3 fold) and KCl (2-fold) but was unaffected by glucose. These results suggest that the HIT T15-G cells may represent a less differentiated form of the parental HIT T15 cell line in which A cell phenotype is dominant but not complete. PMID- 2558931 TI - Stimulus-secretion coupling of arginine-induced insulin release: metabolism of L arginine and L-ornithine in tumoral islet cells. AB - The metabolism of L-arginine and L-ornithine was examined in tumoral islet cells of the RINm5F line and compared to the situation previously characterized in normal rat islets. The maximal velocity of arginase in cell homogenates, as well as either the production of 14C-urea or the steady-state content of 14C-labelled ornithine in intact cells exposed to L-[U-14C]arginine were about one order of magnitude lower in tumoral than normal islet cells. The activity of ornithine glutamate transaminase was similar in both cell types, and this coincided with a comparable rate of 14C-labelled L-glutamate generation by intact cells exposed to L-[1-14C]ornithine. Despite a comparable cell content in 14C-labelled ornithine of normal and tumoral cells exposed to exogenous ornithine, the rate of di- and polyamine generation was about one order of magnitude higher in tumoral than normal islet cells, this coinciding with a much higher activity of ornithine decarboxylase in RINm5F cell than islet homogenates. PMID- 2558933 TI - Diurnal variations in melatonin binding sites in the hamster brain: impact of melatonin. AB - The distribution of 125I-melatonin binding sites in the male Syrian hamster brain was recorded at 3 times over a 24 h period. The binding in the hypothalamus, hippocampus, medulla-pons and midbrain of the hamsters varied significantly over the 24 h period with different patterns and phases. No such variations were observed in the parietal cortex. Daily morning (10.00 h) or late afternoon (18.00 h) injections of melatonin for 28 days markedly increased the serum concentrations of melatonin at all times recorded. Serum concentrations of testosterone were significantly lower in animals injected with melatonin in the late afternoon than in the untreated controls; no such decrease was observed in animals injected in the morning despite the continuously elevated levels of circulating melatonin. The daily melatonin injections did not significantly affect 125I-melatonin binding in the hypothalamus, parietal cortex and medulla pons. In the midbrain, 125I-melatonin binding decreased regardless of the time of injection. In the hippocampus, morning melatonin injections caused a marked decrease in 125I-melatonin binding at all times recorded whereas melatonin injected in the late afternoon led to a decrease in 125I-melatonin binding at 10.00 h only. These results indicate diurnal variations in 125I-melatonin binding sites in discrete brain areas of the golden hamster, persisting despite prolonged duration of elevated levels of circulating melatonin. The differential effects of timed melatonin injections on the hippocampal 125I-melatonin binding sites are positively correlated with the counter-antigonadal response produced by morning melatonin injections. PMID- 2558934 TI - Examination of parathyroid hormone antisera for the presence of receptor antibodies. AB - Proteins from bovine kidney membranes were separated by denaturating polyacrylamide gel electrophoresis and blotted onto nitrocellulose paper. The blots were immunostained with parathyroid hormone (PTH) antisera, and the effect of the presence of PTH on immunostaining was determined. Immunostaining of membrane proteins by two specific antisera was altered by PTH. With one antiserum, the immunostaining of two specific proteins (apparent mass 90 and 105 kDa) was prevented by PTH. With the second antiserum the immunostaining of a 150 kDa protein was prevented by the hormone. These effects were strongest with the 90 and 150 kDa proteins and these were investigated further. Antibody binding was prevented either by co-incubation or by preincubation of the blots with PTH, followed by washing and subsequent exposure to the antisera. Concentrations of PTH as low as 1 nM prevented antibody binding to the 90 kDa species, but somewhat higher PTH concentrations were required with the 150 kDa protein. Oxidation of the PTH methionine residues in the amino terminal segment of PTH, and deletion of the first nine residues in the hormone greatly reduced the competition with the 90 kDa protein, but had no effect on immunostaining of the 150 kDa species. The 35-84 fragment of PTH was not a competitor for the 90 kDa species, while the 1-34 fragment was ineffective with the 150 kDa protein.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2558935 TI - Insulin does not activate a phosphoinositide-specific phospholipase C in adipocytes. AB - Phosphoinositide-specific phospholipase C (PI-PLC) activity was determined in homogenates of adipocytes treated with maximal concentrations of insulin. PI-PLC activity measured using exogenous [3H]phosphatidylinositol [( 3H]PI) and exogenous [3H]phosphatidylinositol 4,5-bisphosphate [( 3H]PIP2) was not altered by prior exposure of adipocytes to insulin. It was possible to see oxytocin induced breakdown of phosphoinositides but no effect of insulin was seen in intact adipocytes. PMID- 2558936 TI - Cyclic AMP-mediated cytoskeletal effects in adrenal cells are modified by serum, insulin, insulin-like growth factor-I, and an antibody against urokinase plasminogen activator. AB - In adrenocortical cells in culture, increased intracellular cyclic AMP resulting from exposure to agents such as ACTH and cholera toxin causes a change in cell morphology termed 'retraction' or 'rounding'. The breakdown of actin-containing stress fibers in rounding suggested a role for microfilaments in steroidogenesis. Previously, we showed that cultured bovine adrenal cells under standard conditions (medium with 10% fetal bovine serum) do not round in response to intracellular cyclic AMP. Here, we show that these cells do round in defined, serum-free medium. Rounding was maximal within 1 h of addition of 1 nM cholera toxin and after 10 h most cells remained rounded. Cycloheximide at 100 micrograms/ml did not inhibit the response to cholera toxin. The rounding response was abolished when 10% fetal bovine serum, horse serum, or ether extracted fetal bovine serum was included in the medium. The inhibitory effect of serum was not mimicked by growth factors with the exception that insulin and insulin-like growth factor-I (IGF-I), while not preventing rounding, accelerated the return of cells to a flattened morphology. A monoclonal antibody against urokinase plasminogen activator completely prevented rounding whereas a monoclonal antibody against tissue plasminogen activator had only a slight effect. Fluorescence visualization of F-actin with N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl)-phallacidin showed that rounding in cultured bovine adrenocortical cells resembles that defined earlier for human and rat adrenocortical cells and includes depolymerization of actin microfilaments. These cytoskeletal changes in adrenal cells are unlikely to play a role in steroidogenesis; however, they may be involved in tissue remodeling occurring as part of the indirect mitogenic effects of ACTH. PMID- 2558938 TI - Interdigestive gastroduodenal motility in patients with active and inactive duodenal ulcer disease. AB - The interdigestive gastroduodenal motility was studied by means of a manometric probe in 6 patients with active duodenal ulcer and acid hypersecretion, in 6 patients with ulcer disease in remission (inactive) and normosecretion and in 8 healthy subjects with normosecretion. After a basal recording period sufficient to record at least two activity fronts of the migrating motor complex (MMC), an intraluminal infusion of isotonic NaHCO3 was carried out for 180 min in patients with active duodenal ulcer, whereas in patients with ulcer in remission an HCl solution was infused for 180 min. Patients with active duodenal ulcer showed a basal motility with a longer than normal MMC cycle and a shorter than normal activity front, while patients with ulcer in remission showed a cyclic motor activity not significantly different from that of normal subjects. The NaHCO3 infusion in patients with active ulcer restored a near-normal motility, whereas the HCl infusion in patients with ulcer in remission induced a motility similar to that of patients with active ulcer. These data indicate that the increase in gastric acid secretion is responsible for the decrease in frequency and duration of MMC activity fronts, which have the function of cyclically clearing the gastroduodenal lumen. Consequently, acid and bacteria may remain a longer than normal time in contact with the gastroduodenal mucosa, which, in this manner, may be greatly exposed to the risk of peptic lesions. PMID- 2558937 TI - Functional receptors for insulin-like growth factors I and II in rat thymocytes and mouse thymoma cells. AB - Functional receptors for insulin-like growth factors (IGF) I and II have been identified in rat thymocytes and mouse thymoma cell lines R1.1 and S49.1. IGF-I receptor alpha-subunit (MW 130,000) bind IGF-I and IGF-II with equal affinity (Kd approximately 4-7 nM), and insulin with approximately 100 times lower affinity. Tyrosine kinase activity and autophosphorylation of the IGF-I receptor beta subunit (MW 95,000) are stimulated by IGF-I and IGF-II with equal potency (ED50 approximately 0.5 nM). IGF-II receptors (MW 250,000) bind IGF-II with Kd approximately 0.3 nM and IGF-I with 30 times lower affinity, but not insulin. IGF I and IGF-II do not cross-react with the insulin receptor to which insulin binds with an apparent Kd approximately 1 nM, and stimulates its tyrosine kinase activity with ED50 approximately 3 nM. In thymocytes, alpha-aminoisobutyric acid transport is stimulated 2-fold by IGF-I and IGF-II with identical potency (ED50 approximately 2 nM), and by insulin with ED50 approximately 10 nM. Activation of thymocytes by concanavalin A increased the number of IGF-II receptors 2-fold, whereas IGF-I receptor binding and IGF-stimulated amino acid transport were unaltered. We conclude that the effect of IGF-I and IGF-II in thymocytes is mediated via binding to the IGF-I receptor and stimulation of its tyrosine kinase. The presence of functional IGF receptors on thymocytes and thymoma cells suggests that IGF-I and IGF-II play a role in the regulation of thymic functions. PMID- 2558939 TI - 5'-flanking DNA of the human insulin receptor gene and long terminal repeat of mouse mammary tumour virus bind to the same nuclear protein(s). AB - The interaction of nuclear protein extracted from rat liver and 5'-flanking DNA of the human insulin receptor gene was investigated with the aid of gel mobility shift analysis. When 5'-flanking DNA (-1255/-1206 or -385/-345 base pairs) was incubated with nuclear protein, two or three 32P-DNA species (protein binding DNA fragment(s) and free DNA fragment) were detected. These bands did not disappear in spite of increasing amounts of synthetic poly(dI-dC), showing that nuclear protein binds specifically to 5'-flanking DNA of the insulin receptor gene. Increasing amounts of long terminal repeat of mouse mammary tumour virus resulted in a reciprocal decrease in nuclear protein binding to 5'-flanking DNA of insulin receptor gene. These results suggest that 5'-flanking DNA of insulin receptor gene binds to the same nuclear protein to which long terminal repeat of mouse mammary tumour binds. PMID- 2558940 TI - Lamellar body formation precedes pulmonary surfactant apoprotein expression during embryonic mouse lung development in vivo and in vitro. AB - The purpose of this investigation was to determine whether lamellar inclusion body (LB) formation and surfactant apoprotein (SP-35) production are directly coordinated by temporal and positional information during development. In the present study we report a comparison between embryonic B10.A mouse lung morphogenesis and cytodifferentiation in vivo with that observed during organ culture in serumless medium. Precursor LB were first detected at embryonic day 12 (E12d), and progressively larger numbers and forms were produced during subsequent differentiation of respiratory alveolar duct epithelium. SP-35 was first detected during the canalicular period (E16.5d). Lung cultures (E12d) showed pseudoglandular and canalicular periods of morphogenesis, and both ciliated epithelial and type II cell differentiation. Nonciliated cells produced increasing numbers of lamellar inclusion bodies throughout the culture period. SP 35 was detected at 9 days in vitro (d.i.v.). These observations indicate (i) precursor LB formation precedes SP-35 expression and is not dependent on apoprotein synthesis; (ii) E12d lung development in vitro using serumless medium proceeds at a rate equivalent to 0.5 days in vivo through 11 d.i.v.; and (iii) morphogenesis and differentiation occur in the absence of exogenous hormones and growth factors. The cell-cell interactions that play a role in morphogenesis and cell differentiation appear to be intrinsic to the developmental program for embryonic lung development and are likely to be mediated by autocrine and/or paracrine factors. PMID- 2558942 TI - Induction of ornithine decarboxylase in the stomach mucosa of swine with NaCl or 12-O-tetradecanoylphorbol-13-acetate. AB - The effect of sodium chloride and 12-O-tetradecanoylphorbol-13-acetate on ornithine decarboxylase (ODC) activity in gastric mucosa of miniature swine was investigated as a model for gastric inflammation. The level of the enzyme was lower in the pylorus than in the fundic or cardiac regions of the stomach in untreated animals. Treatment with sodium chloride at 1 g/kg produced large increases in all three regions, with the greatest relative increase in the pylorus. Treatment with sodium chloride at 0.25 g/kg or the phorbol ester at 2.0 mg/pig produced significant but less dramatic increases. ODC activity in control and treated mucosal extracts was inhibited by the specific ODC inhibitor difluoromethylornithine. Most of the enzyme activity was associated with superficial and exfoliated cells that could be scraped from the mucosal surface. No increase in the inflammatory mediator leukotriene B4 was observed in the mucosal extracts. Ornithine decarboxylase appears to be a useful enzymatic marker for the regenerative events that occur after tissue damage and may correlate with the putative tumor-promoting function of sodium chloride in gastric tissues. PMID- 2558941 TI - Transforming growth factor-beta and its receptor are differentially regulated in human embryonal carcinoma cells. AB - The human embryonal carcinoma cell lines Tera-2 clone 13 and NTera-2 clone D1 can be induced by retinoic acid to differentiate in vitro into neuroectodermal derivatives. The undifferentiated cells are rapidly proliferating and tumorigenic, whereas retinoic-acid-treated cells possess a decreased growth rate, lose their transformed phenotype and show a finite lifespan. Differentiation is accompanied by a marked increase in the levels of mRNA for TGF-beta 1 and TGF beta 2 and the production of TGF-beta activity. Just like murine embryonal carcinoma cells the growth of Tera-2 clone 13 cells is not affected by the addition of either TGF-beta 1 or TGF-beta 2 to the culture medium. In contrast to published data on murine embryonal carcinoma cells, Tera-2 clone 13 and NTera-2 clone D1 cells bind TGF-beta 1 with high affinity, which is due to the presence of type-III TGF-beta receptors. Furthermore, and again in contrast to murine embryonal carcinoma cells, treatment of the human embryonal carcinoma cells with retinoic acid causes a nearly complete loss of TGF-beta 1 binding sites. These results are discussed in the light of similarities and differences in the regulation of growth and differentiation of human and murine embryonal carcinoma cell lines. PMID- 2558943 TI - [Cryptococcus neoformans meningitis and cirrhosis. Value of fluconazole]. AB - We report the case of a 80-old-year woman with cryptococcal meningitis and cirrhosis. The diagnosis of this rare infectious complication is probably underestimated in patients with cirrhosis. Of diagnostic value is the detection of cryptococcal antigen in cerebral fluid. Fluconazole could improve an otherwise poor prognostic. PMID- 2558945 TI - Investigations of excitability of isolated and non-isolated neurons from the terminal ganglion of Periplaneta americana by current/voltage clamp and intracellular perfusion. AB - It was shown by means of current and voltage clamp measurements in combination with the intracellular perfusion method that the isolated motoneuron nerve cell bodies of the cockroach Periplaneta americana show overshoot action potentials. A regenerative Na current was found to be responsible for the excitability of these membranes; however, the results are not in agreement with a de novo synthesis of these channels. PMID- 2558944 TI - [Long-term studies of patients with borderline tumors of the ovaries]. AB - This retrospective investigation comprises a follow-up observation of 51 patients with borderline tumours of the ovaries, who had been treated at the Department of Gynaecology of the University of Erlangen between January 1st 1966 and December 31st 1986. The average age was approx. ten years younger that of patients with invasive carcinomas and 65% of the patients were in stage I. For women in stage Ia desiring children, unilateral adnexectomy is sufficient in the context of meticulous staging laparotomy. The remaining ovary should be further exposed by wedge excision. Once the family is complete, hysterectomy and adnexectomy should then be carried out retrospectively in view of the high risk of bilaterality. In ovarian tumours of borderline malignancy stages II and III, the surgical therapy should correspond to the procedure for invasive carcinomas, with the objective of postoperative freedom from tumour. Lymphonodectomy is not obligatory in view of the very rare involvement of lymph nodes. The question as to the necessity of adjuvant therapy is still open. Our observations indicate, that combination chemotherapy, containing cisplatinum is effective as adjuvant therapy in advanced stages as well as in recurrences, which cannot be completely resected surgically. 18% of our patients with borderline tumours suffered a recurrence or tumour progression, in consequence of which five died (10%). The recurrences were manifested clinically after 33 months at the earliest, and after 164 months at the latest, so that the follow-up in these patients should extend over a period of more than ten years. PMID- 2558946 TI - The interaction of fructose-1,6-biphosphate aldolase with liposome membranes: a spin probe technique study. AB - Thermotropic properties of liposome membranes prepared of bulk bovine erythrocyte membrane lipids, native, or aldolase-modified, were investigated by the ESR method. Breaks were observed in the log 2T parallel vs 1/T plots for two spin labels: tempopalmitate and 5-doxyl-palmitate methyl ester. These phenomena have been interpreted as reflecting structural changes near the lipid bilayer polar heads region. Upon modification with aldolase, the temperature at which the breaks occurred was decreased for both spin probes. PMID- 2558947 TI - Functional characteristics of phosphatidylinositol-specific phospholipases C from Bacillus cereus and Bacillus thuringiensis. AB - Phosphatidylinositol-specific phospholipase C was purified from the culture medium of B. thuringiensis to high specific activity using a procedure we recently described for purification of PI-PLC from B. cereus (Volwerk et al. (1989) J. Cell. Biochem. 39, 315-325). The purified enzymes from B. thuringiensis and B. cereus have similar specific activities towards hydrolysis of the membrane lipid phosphatidylinositol, and also towards hydrolysis of the glycosyl phosphatidylinositol-containing membrane anchor of bovine erythrocyte acetylcholinesterase. These results indicate very similar catalytic properties for the structurally homologous PI-specific phospholipases C secreted by these bacilli. PMID- 2558948 TI - The outer membrane protein of enteropathogenic Escherichia coli, described as the 'localised adherence factor', is OmpF and probably not involved in adhesion to HEp-2 cells. AB - Strains of enteropathogenic Escherichia coli (EPEC) were examined for a factor, described as an outer membrane protein (OMP) of 32 kilodaltons (kDa) and reported to be involved in the adhesion of EPEC to HeLa cells. A comparable OMP of 35 kDa was detected in strains of EPEC, although expression of this protein was not related to the ability of strains to adhere to HEp-2 cells. The 35 kDa OMP was found to be heat-modifiable and peptidoglycan associated, and considered to be the porin protein OmpF. PMID- 2558949 TI - Mutations affecting hemolysin production in Listeria monocytogenes located outside the listeriolysin gene. AB - We have investigated the molecular basis of spontaneous mutations leading to non hemolytic and avirulent variants of the Listeria monocytogenes serotype 1/2a strain NCTC 7973 using Southern hybridization to DNA fragments that harbor the listeriolysin gene (hlyA) and adjacent regions cloned from a L. monocytogenes serotype 1/2a strain. The analysis of such non-hemolytic variants revealed the presence of a deletion of 300 base pairs, located 1.6 kb upstream of an otherwise intact listeriolysin gene. The importance of regions upstream of the hlyA gene in controlling the expression of the listeriolysin gene was further emphasized by the detection of a transposon-derived nonhemolytic mutant in which the transposon had inserted approximately 200 bp upstream of the listeriolysin gene. We conclude that at least two elements, contained within a region encompassing 1.6 kb of sequences upstream of the hlyA gene, may be required for expression of the listeriolysin gene. PMID- 2558950 TI - Increased expression of biodegradative threonine dehydratase of Escherichia coli by DNA gyrase inhibitors. AB - The synthesis of inducible biodegradative threonine dehydratase of Escherichia coli increased several-fold in the presence of the DNA gyrase inhibitors, nalidixic acid and coumermycin. Temperature-sensitive gyrB mutants expressed higher levels of dehydratase as compared to an isogenic gyrB+ strain. Immunoblotting experiments showed increased synthesis of the dehydratase protein in the presence of gyrase inhibitors; addition of rifampicin and chloramphenicol to cells actively synthesizing enzyme preventing new enzyme production. Increased expression of dehydratase by gyrase inhibitors was accompanied by relaxation of supercoiled DNA. PMID- 2558951 TI - Expression of foreign DNA in Chlamydomonas reinhardtii. AB - A chimeric octopine synthase-neomycin phosphotransferase (ocs-nptII) gene was used to transform Chlamydomonas reinhardiii to kanamycin resistance. Southern hybridization using DNA isolated from one transformant, T6.1, indicated that the entire ocs-nptII gene and at least part of the plasmid were integrated into nuclear DNA. Neomycin phosphotransferase II activity has been detected in T6.1 cell extracts. Northern hybridizations, employing a radiolabeled ocs-nptII sequence, revealed a T6.1 transcript of approximately the same size as a homologous transcript isolated from E. coli carrying the nptII gene. Although T6.1 is an extremely rare example of a stable C. reinhardtii transformant, its occurrence nevertheless indicates that bacterial genes can be expressed in the nucleus of the alga. PMID- 2558952 TI - Identification of the rhaA, rhaB and rhaD gene products from Escherichia coli K 12. AB - The three structural genes rhaA, rhaB and rhaD, that specify the enzymes rhamnose isomerase, rhamnulose kinase and rhamnulose 1-phosphate aldolase respectively, have been cloned from Escherichia coli K-12. The precise location of the genes has been determined by gene complementation analysis and by enzymatic assays of strains transformed with recombinant plasmids containing different parts of the cloned region. The corresponding gene products have been studied by their expression in maxicells. Protein products of 47 kDa, 52-54 kDa and 32 kDa have been assigned to rhamnose isomerase, rhamnulose kinase and rhamnulose 1-phosphate aldolase respectively. PMID- 2558953 TI - Cloning of the virulence regulatory (vir) locus of Bordetella pertussis and its expression in B. bronchiseptica. AB - Hybridisation with cosmid pRMB2, containing the vir locus of bordetella pertussis, showed that the Tn5 insertion in B. pertussis BP347 (Vir-) was located with 1 2.7 kB EcoRI fragment. When subcloned, this fragment alone was unable to complement BP347, but a larger 8.0 kb region which included the 2.7 kb EcoRI fragment, did restore expression of virulence associated properties to BP347, and to avirulent phase variants of both B. pertussis and B. bronchiseptica. EcoRI digested DNA from stains of all four species of Bordetella showed homology to the cosmid genomic insert and to the 2.7 kb subclone, but B. avium showed a markedly different pattern of homology to the other species. PMID- 2558954 TI - Genetic analysis of temperature-sensitive lethal mutants of Salmonella typhimurium. AB - We have isolated 440 mutants of Salmonella typhimurium that show temperature sensitive growth on complex medium at 44 degrees. Approximately 16% of the mutations in these strains have been mapped to 17 chromosomal locations; two of these chromosomal locations seem to include several essential genes. Genetic analysis of the mutations suggests that the collection saturates the genes readily mutable to a ts lethal phenotype in S. typhimurium. Physiological characteristics of the ts lethal mutants were tested: 6% of the mutants can grow at high temperature under anaerobic conditions, 17% can grow when the medium includes 0.5 M KCl, and 9% of the mutants die after a 2-hr incubation at the nonpermissive temperature. Most ts lethal mutations in this collection probably affect genes required for growth at all temperatures (not merely during high temperature growth) since Tn10 insertions that cause a temperature-sensitive lethal phenotype are rare. PMID- 2558955 TI - Use of pulsed field gel electrophoresis and transposon mutagenesis to estimate the minimal number of genes required for motility in Caulobacter crescentus. AB - To facilitate the mapping of transposon insertion mutations in Caulobacter crescentus, we have used pulsed field gel electrophoresis to construct a detailed physical and genetic map of the C. crescentus genome. Restriction fragments were generated by DraI, AseI, or SpeI which cleave the C. crescentus 40, 13, and 26 times, respectively, and Tn5 insertions were used to align the restriction fragments generated by each of the enzymes. The utility of the resulting map was demonstrated by determining the chromosomal locations of a collection of flagellar mutations. As a result of this study, we were able to identify ten new flagellar genes at various locations on the chromosome. Thus, at least 48 genes are required for the assembly of a functional flagellum in C. crescentus. PMID- 2558957 TI - Yeast intrachromosomal recombination: long gene conversion tracts are preferentially associated with reciprocal exchange and require the RAD1 and RAD3 gene products. AB - A yeast intrachromosomal recombination system based on an inverted repeat has been designed to examine mitotic gene conversion tract length and the association of crossing over with gene conversion as a function of the conversion tract length. Short conversion tracts are found to be preferentially noncrossover while conversion tracts longer than 1.16 kb show a 50% association with crossover. Mutation in the excision repair gene RAD1 leads to a reduction in conversion tracts of at least 1.16 kb and a reduction in crossovers associated with conversion, regardless of the length of the conversion tract. Mutation in the excision repair gene RAD3, which encodes a DNA helicase, also leads to a reduction in conversion tracts of at least 1.16 kb, but has no effect on the frequency of associated crossovers. The roles of RAD1 and RAD3 in recombination are discussed. PMID- 2558956 TI - Analysis of yeast retrotransposon Ty insertions at the CAN1 locus. AB - The target site distribution for 55 independent Ty insertions that inactivate the function of the Saccharomyces cerevisiae CAN1 gene is reported. Under some selection conditions Ty elements inserted preferentially into the promoter and exhibited an orientation bias. In contrast, under other conditions no insertions were detected in the promoter region and transposition appeared to occur randomly throughout the CAN1 coding sequence. These results show that the target site distribution for Ty insertions may be a function of the selection conditions. PMID- 2558959 TI - Modified P elements that mimic the P cytotype in Drosophila melanogaster. AB - Activity of the P family of transposable elements in Drosophila melanogaster is regulated primarily by a cellular condition known as P cytotype. It has been hypothesized that P cytotype depends on a P element-encoded repressor of transposition and excision. We provide evidence in support of this idea by showing that two modified P elements, each with lesions affecting the fourth transposase exon, mimic most of the P cytotype effects. These elements were identified by means of two sensitive assays capable of detecting repression by a single P element. One assay makes use of cytotype-dependent gene expression of certain P element insertion mutations at the singed bristle locus. The other measures suppression of transposase activity from the unusually stable genomic P element, delta 2-3(99B), that normally produces transposase in both germinal and somatic tissues. The P cytotype-like effects include suppression of snw germline hypermutability, snw somatic mosaicism, pupal lethality, and gonadal dysgenic sterility. Unlike P cytotype, however, there was no reciprocal cross effect in the inheritance of repression. PMID- 2558960 TI - Somatic reversion of P transposable element insertion mutations in the singed locus of Drosophila melanogaster requiring specific P insertions and a trans- acting factor. AB - Destabilization in somatic cells of P-element insertions in the X-linked singed gene of Drosophila melanogaster has been studied. We have shown that some but not all unstable P-element insertions in singed can form mosaics. The cause of this variation is not clear from studies of the restriction maps of the mutations tested. The transposable element movements occur early in development and require, in addition to an appropriate P-element insertion in singed, a trans acting maternal effect component. Movements appear to occur preferentially in attached-X stocks. However, the maternal effect component maps to the central region of chromosome 2. PMID- 2558958 TI - Isolation and characterization of temperature-sensitive mutations in the RAS2 and CYR1 genes of Saccharomyces cerevisiae. AB - The yeast Saccharomyces cerevisiae contains two ras homologues, RAS1 and RAS2, whose products have been shown to modulate the activity of adenylate cyclase encoded by the CYR1 gene. To isolate temperature-sensitive mutations in the RAS2 gene, we constructed a plasmid carrying a RAS2 gene whose expression is under the control of the galactose-inducible GAL1 promoter. A ras1 strain transformed with this plasmid was subjected to ethyl methanesulfonate mutagenesis and nystatin enrichment. Screening of approximately 13,000 mutagenized colonies for galactose dependent growth at a high temperature (37 degrees) yielded six temperature sensitive ras2 (ras2ts) mutations and one temperature-sensitive cyr1 (cyr1ts) mutation that can be suppressed by overexpression or increased dosage of RAS2. Some ras2ts mutations were shown to be suppressed by an extra copy of CYR1. Therefore increased dosage of either RAS2 or CYR1 can suppress the temperature sensitivity caused by a mutation in the other. ras1 ras2ts and ras1 cyr1ts mutants arrested in the G1 phase of the cell cycle at the restrictive temperature, and showed pleiotropic phenotypes to varying degrees even at a temperature permissive for growth (25 degrees), including slow growth, sporulation on rich media, increased accumulation of glycogen, impaired growth on nonfermentable carbon sources, heat-shock resistance, impaired growth on low concentrations of glucose, and lithium sensitivity. Of these, impaired growth on low concentrations of glucose and sensitivity to lithium are new phenotypes, which have not been reported for mutants defective in the cAMP pathway. PMID- 2558961 TI - A study of ten families of transposable elements on X chromosomes from a population of Drosophila melanogaster. AB - Data were collected on the distribution of ten families of transposable elements among fourteen X chromosomes isolated from a natural population of Drosophila melanogaster, by means of in situ hybridization to polytene chromosomes. It was found that, with the exception of roo, the copy number per chromosome followed a Poisson distribution. There was no evidence for linkage disequilibrium, either within or between families. Some pairs of families of elements were correlated with respect to the identity of the sites that were occupied in the sample, although there was no evidence for a correlation with respect to the sites at which elements attained relatively high frequencies. Elements appeared to be distributed randomly along the distal part of the X chromosome. There was, however, a strong tendency for elements to accumulate at the base of the chromosome. Element frequencies per chromosome band were generally low, except at the base of the chromosome where bands in subdivisions 19E and 20A sometimes had high frequencies of occupation. These results are discussed in the light of models of the population dynamics of transposable elements. It is concluded that they provide strong evidence for the operation of a force or forces opposing transpositional increase in copy number. The accumulation of elements at the base of the chromosome is consistent with the idea that unequal exchange between elements at non-homologous sites is such a force, although other possibilities cannot be excluded at present. The data suggest that the rate of transposition per element per generation is of the order of 10(-4), for the elements included in this study. PMID- 2558962 TI - The occurrence of long ribosomal transcripts homologous to type I insertions in bobbed mutants of Drosophila melanogaster. AB - In Drosophila melanogaster up to two thirds of the rDNA genes contain insertion sequences of two types in the 28S coding region. Comparison of the ribosomal insertion transcripts in the wild type and in two bobbed mutants reared at two temperatures showed that the level of type I transcripts is dependent on both the number of genes with type I insertions in the bobbed loci and the intensity of bobbed phenotype. Importantly, a long transcript of 8.7 kb hybridized to the ribosomal probe, the INS I probe and also to the restriction fragment of the rDNA downstream of the point of insertion was found in one bobbed mutant. This result and also those from sandwich hybridization indicate that some interrupted ribosomal genes are functional. PMID- 2558963 TI - P-element transposase induces male recombination in Drosophila melanogaster. AB - Male recombination in P-M dysgenic crosses has been viewed as a reflection of P element transposase interacting with P elements. However, recent studies suggest that the transposase may catalyse double-stranded breaks in chromosomal DNA. We have, therefore, introduced P(delta 2-3 ry+) (99B), a single non-mobile P-element transposase source, into the long-standing laboratory true M strains of a flanking lethal crossover selective system, thus facilitating the examination of rare male recombination events as an assay for transposase activity. We find that the rate of male recombination in the presence of this non-mobile P element is greater than twenty times the background rate of male recombination in the control examined prior to introduction of the transposase source. PMID- 2558964 TI - Origin of rare Ha-ras alleles: relationship of VTR length to a 5' polymorphic Xho I site. AB - Amongst the four common Ha-ras alleles in both controls and cancer patients, we detected the presence of a polymorphic Xho I site associated specifically with the 6.6 and 7.7 kb Bam HI fragments but absent from the 7.1 and 8.2 kb alleles, as recently reported by others. We have extended this study and report here, the consistent appearance of this Xho I site in unusual alleles close in size to the two common alleles of 6.6 and 7.7 kb, in control lymphoblastoid DNA samples in a variety of tumor DNAs. Unusual alleles grouped around the 7.1 and 8.2 kb common alleles on the other hand, did not possess the Xho I site. The consistent presence of the Xho I site polymorphism, in the unusual Ha-ras alleles surrounding the 6.6 and 7.7 kb common alleles and its absence in alleles around the 7.1 and 8.2 kb common alleles, suggests that the unusual ones are derived from the corresponding common alleles to which they are closest in size. PMID- 2558965 TI - The need for enhancers in gene expression first appears during mouse development with formation of the zygotic nucleus. AB - Microinjection of the firefly luciferase gene coupled to a thymidine kinase (tk) promoter provided a quantitative assay to evaluate the requirements for gene expression in individual mouse oocytes and embryos. Polyoma virus (PyV) enhancers had no effect on the level of gene expression or competition for transcription factors as long as the DNA remained either in the oocyte germinal vesicle or the pronuclei of one-cell embryos. Expression of injected genes could be observed in pronuclei because the signal that normally triggers zygotic gene expression in two-cell embryos still occurred in one-cell embryos arrested in S phase. However, when the tk promoter was injected into zygotic nuclei of two-cell embryos, enhancers increased the number of embryos that expressed luciferase as well as the level of luciferase activity per embryo. PyV enhancer mutation F101, selected for growth in mouse embryonal carcinoma F9 cells, stimulated expression in developing two-cell embryos about seven times better than the wild-type PyV enhancer and competed effectively for factors required for transcription. These results were consistent with the fact that enhancers are required to activate the PyV origin of DNA replication in developing two-cell embryos but not in one-cell embryos. The maximum levels of gene expression in oocytes, one-cell embryos, and developing two-cell embryos (1:67:21) were inversely related to the extent of chromatin assembly, but the need for enhancers was independent of chromatin assembly. Therefore, it appears that the need for enhancers to activate promoters or origins of replication results from some negative regulatory factor that first appears as a component of zygotic nuclear structure. PMID- 2558967 TI - The nature of an intragenic suppressor of the Escherichia coli dnaA508 temperature-sensitive mutation. AB - Escherichia coli strain E508 (dnaA508) is temperature-sensitive for dnaA function. A mutant with an intragenic suppressor of the dnaA508 mutation, called PR1, has been isolated. The suppressor mutation(s) allow initiation of DNA synthesis at 42 degrees C and, like dnaA cold-sensitive mutants, PR1 grows poorly at 32 degrees C. Two-dimensional gel analysis indicates that DnaA protein is overproduced in PR1. Transcriptional analysis indicates two to three times the number of dnaA and dnaN transcripts in PR1, as compared to a wild-type dnaA+ strain. The dnaA gene from PR1 has been cloned and found to complement the original dnaA508 mutation, as well as dnaA46, but not dnaA5. Sequencing of the dnaAPR1 gene reveals three separate base changes, two of which result in nonconservative amino acid substitutions and the third is a change in the start codon from GTG to ATG. PMID- 2558966 TI - Mammalian pre-mRNA branch site selection by U2 snRNP involves base pairing. AB - SV40 early pre-mRNA is alternatively spliced to produce large T and small t mRNAs by use of different 5'-splice sites and a shared 3'-splice site. The large T splicing pathway uses multiple lariat branch sites, whereas small t splicing, constrained by its small intron size, can use only one. We exploited this situation to test the hypothesis that RNA-RNA base pairing between U2 snRNA and the branch site sequence is important in mammalian pre-mRNA splicing by constructing and analyzing several mutations in the small t pre-mRNA branch site (UUCUAAU). All of the mutations resulted in substantial decreases in small t splicing relative to large T. To test whether these effects resulted from decreased base pairing with U2 snRNA, compensatory mutations were introduced at the appropriate positions (nucleotides 34-36) in a cloned human U2 gene. All branch site mutations tested (four separate single base substitutions and two triple mutations) were suppressed (i.e., small t splicing was increased) by the appropriate U2 mutations. These results establish that recognition of the poorly conserved mammalian pre-mRNA branch site sequence by U2 snRNP can involve base pairing. PMID- 2558968 TI - Molecular cloning and in vitro expression of a silent phenoxazinone synthase gene from Streptomyces lividans. AB - Phenoxazinone synthase (PHS) catalyzes a step in actinomycin D biosynthesis in Streptomyces antibioticus. Two sequences from Streptomyces lividans that hybridize to the phs gene of S. antibioticus have been cloned in Escherichia coli K-12 using the plasmid pBR322. Although there was some similarity in the restriction maps of the two cloned fragments, neither insert appeared to be a direct subset of the other nor of the S. antibioticus phs gene. In vitro expression studies, in a streptomycete coupled transcription-translation system, showed that a 3.98-kb SphI fragment encoded a PHS-related protein. These observations provide additional support for the existence of silent genes for antibiotic production in streptomycetes. PMID- 2558969 TI - Cloning of a new bidirectionally selectable marker for Aspergillus strains. AB - Mutants that lack adenosine triphosphate sulfurylase (ATPsase; EC 2.7.7.4) are unable to use sulfate as sole source of sulfur and are also resistant to selenate. These mutants, denoted sC-, are readily obtained from any strain of Aspergillus niger or Aspergillus nidulans by the strong selection for selenate resistance. We have cloned the gene encoding ATPsase from A. nidulans by complementation of an sC mutant strain of A. nidulans with a gene library and show that plasmids containing this gene transform both A. niger and A. nidulans sC- strains, restoring their ability to grow on sulfate as sole sulfur source. The fact that strong selection for either sC+ or sC- can be applied provides a simple way of delivering genetically engineered constructs to any strain of A. niger including strains of industrial importance. In addition, this system is useful for gene replacements and other genomic DNA manipulations in Aspergillus species. PMID- 2558970 TI - Rapid identification of DNA fragments containing promoters for RNA polymerase II. AB - We describe a direct procedure for screening genomic recombinant DNA libraries or restriction fragments of cloned DNA regions for RNA polymerase II promoters. Cellular polyadenylated mRNA is chemically de-capped by beta-elimination reaction and enzymatically re-capped with [alpha-32P]GTP by vaccinia guanylyl transferase. Since this enzyme only accepts di- or triphosphorylated 5' termini as a substrate, the mRNAs are labeled exclusively at the first nucleotide, irrespective of whether the mRNA was intact or fragmented before in vitro capping. By using in vitro-capped mRNA as a hybridization probe, recombinant DNA molecules or restriction fragments that carry a cap site (and thus likely an RNA polymerase II promoter) can directly be identified. Here, we demonstrate the applicability of this procedure by the isolation and characterization of several genomic DNA clones containing RNA polymerase II promoter sequences, that are highly active in liver. PMID- 2558971 TI - An auto-inducible vector conferring high glucocorticoid inducibility upon stable transformant cells. AB - A new gene expression system in mammalian cells was developed by using the glucocorticoid receptor (GR) as an inducible positive feedback factor. Mouse Ltk- cells were transfected with plasmids carrying the GR-encoding gene and the lacZ reporter gene, both of which were fused with the glucocorticoid-inducible enhancer/promotor of the mouse mammary tumor virus (MTV). The GR gene was first induced to supply the receptor protein, which further induced the expression of both GR and reporter genes. Stable transformants induced with dexamethasone, a synthetic glucocorticoid hormone, demonstrated beta-galactosidase activity 60-140 fold higher than uninduced controls. Similarly, the human alpha-interferon encoding gene fused with the MTV enhancer/promoter was induced more than 12,000 fold. This system allowed us to increase the expression of the reporter or target genes without augmenting basal levels of expression significantly, and may be useful to investigate the unknown function of a cloned gene, particularly when the gene product of interest is cytotoxic or growth-inhibiting. PMID- 2558972 TI - Homologies between herpesvirus of turkey and Marek's disease virus type-1 DNAs within two co-linearly arranged open reading frames, one encoding glycoprotein A. AB - The genomes of two avian herpesviruses, Marek's disease virus type 1 (MDV1) and herpesvirus of turkey (HVT), share close homology only within certain DNA regions. One such homologous region of HVT DNA was cloned and sequenced. Two open reading frames (ORFs) were found in the long unique region, ORF1 encoding the glycoprotein A (gA), and ORF2 encoding a still unidentified protein. These two HVT-ORFs are located at almost the same positions as the homologous MDV1-ORFs. The nucleotide sequence homologies between HVT and MDV1 were 73% and 68% for ORF1 and ORF2, respectively. Both the 5'- and 3'-noncoding regions, however, are less conserved. The third letter within every codon of ORF1 and ORF2 showed a mismatch of greater than 50% between the two viruses. The amino acid (aa) sequence homologies between the corresponding putative viral proteins are 83% and 80% for ORF1 (gA) and ORF2, respectively. More than 90% homology was observed in the C terminal region of ORF1 (gA). Furthermore, the deduced aa sequences for both of the ORFs in these two viruses showed considerable homology to two adjoining genes in herpes simplex virus type 1, the glycoprotein C and UL45 genes. PMID- 2558973 TI - Chimerasome-mediated gene transfer in vitro and in vivo. AB - Proteoliposome delivery vesicles can be prepared by the protein-cochleate method [Gould-Fogerite and Mannino, Anal. Biochem. 148 (1985) 15-25; Mannino and Gould Fogerite, Biotechniques 6 (1988) 682-690]. Proteins which mediate the entry of enveloped viruses into cells are integrated in the lipid bilayer, and materials are encapsulated at high efficiency within the aqueous interior of these vesicles. We describe proteoliposome-mediated delivery of proteins and drugs into entire populations of cells in culture. Material can be delivered gradually by Sendai-virus-glycoprotein-containing proteoliposomes. Alternatively, synchronous delivery to a population can be achieved by exposing cell-bound influenza glycoprotein vesicles briefly to low pH buffer. When DNA is encapsulated, chimeric proteoliposome gene-transfer vesicles (chimerasomes), which mediate high efficiency gene transfer in vitro and in vivo, are produced. Stable expression of a bovine papilloma virus-based plasmid in tissue-cultured cells, at 100,000 times greater efficiency than Ca.phosphate precipitation of DNA, with respect to the quantity of DNA used, has been achieved. Stable gene transfer and expression in mice has been obtained by subcutaneous injection of chimerasomes containing a plasmid expressing the early region of polyoma virus. In one experimental group, 50% of the mice developed tumors which were shown to express polyoma virus early proteins and contain the transferred DNA. This is the first report of stable gene transfer in animals mediated by a liposome- or proteoliposome-based system. PMID- 2558974 TI - A transcriptionally regulated expression vector for the fission yeast Schizosaccharomyces pombe. AB - An expression vector for the fission yeast Schizosaccharomyces pombe is described. The vector is designed to facilitate the construction of transcriptional fusions to the promoter of the S. pombe fructose bisphosphatase gene. Transcription from this promoter is regulated by glucose repression over a range of greater than 100-fold. The tight regulation by this promoter should allow for the maintenance of genes whose products are lethal to S. pombe and for the high level production of their protein or RNA products. Intermediate levels of expression can also be achieved by growth on different carbon sources. PMID- 2558975 TI - The human serum amyloid A (SAA)-encoding gene GSAA1: nucleotide sequence and possible autocrine-collagenase-inducer function. AB - We have determined the genomic sequence of the human GSAA1 gene, a member of the family of acute-phase human serum amyloid A (SAA)-encoding genes. This sequence predicts a mature protein of 104 amino acids (aa), several of which differ from residues usually conserved in the sequence of SAA proteins isolated from serum. Despite coding differences, however, the four-exon structure of GSAA1 resembles that of other SAA genes in humans and mice. The N-terminal 25 aa of the mature GSAA1 protein are virtually identical to those of an 'SAA-like' autocrine collagenase inducer produced by rabbit synovial fibroblasts; the latter also differ from the corresponding aa found in SAA in serum. We propose that GSAA1 is the human gene coding for a protein closely related to the SAA, but which is adapted to this important autocrine cytokine function. PMID- 2558976 TI - [Pathogenetic characteristics of diseases of the peripheral nervous system in workers having contact with chromium compounds]. AB - When analyzing the results of clinical neurologic, neurophysiologic, psychologic and laboratory instrumental study of 105 chromium-exposed patients with lumbosacral diseases of the peripheric nervous system there were detected clinical and pathogenetic characteristics of this pathology: distinct manifestations of segmental (homolateral) and generalized vegetovascular disorders, deficiency of sympathetic activation, emotional shifts accompanied by disquieting, depressive and hypochondriac tendencies, elevation of the cerebrovascular tension, body sensitization to chromium, increase in its blood serum and urine content. The account of these characteristics is very important for the improvement of the diagnostic process and raising efficacy of treatment of diseases of the peripheric nervous system in chromium-exposed patients. PMID- 2558977 TI - [Determination of potential carcinogenicity of nickel-containing dust using a micronuclear test]. AB - Mutagenic activity of 3 samples of nickel-containing dusts was experimentally studied in white random-bred rats. Quantity of micronuclei in polychromatophilic erythrocytes of bone marrow smear depended on nickel dust content. Micronuclear test could be used as a rapid method for the determination of potential carcinogenic activity of polymetallic nickel-containing dusts and also as a factor confirming oncologic hazard of chemical substances combined with epidemiologic studies and chronic experiment on animals. PMID- 2558978 TI - Superoxide anion radical-independent pathway for reduction of tetrazolium salts in aerobic mixtures consisting of NADH and 5-methylphenazinium methyl sulfate in the presence of aqueous micelles of nonionic and cationic detergents. AB - The effect of Triton X-100 and certain other nonionic as well as cationic detergents on 5-methyl-phenazinium methyl sulfate (PMS)-mediated reduction of tetrazolium salts was studied under aerobic conditions using an exogenous source of reducing equivalents, such as NADH or by generating NADPH through an enzymatic reaction. In the absence of detergents, 5,10-dihydro-5-methylphenazine (MPH), formed on reduction of 5-methylphenazinium cation (MP+) of PMS by NAD[P]H, was reoxidized allowing first the univalent reduction of molecular oxygen (O2) to the superoxide anion radical (O2-.) which, in turn, reduced tetrazolium salts. In the presence of detergents, however, a significant fraction of the PMS-mediated reduction of tetrazolium salts appeared to proceed without the intervention of O2 . The reasons for this were examined experimentally and it was suggested that the reduced phenazine (i.e., MPH), which is sparingly soluble in aqueous solutions, migrates into detergent micelles where tetrazolium salts are reduced in preference to O2. By lowering the pH and thereby facilitating the H+-mediated dismutation of O2-., it was possible to obtain the reduction of tetrazolium salts, mediated selectively and directly by MPH in the micellar pseudophase. Employing the technique of saturation analysis, further evidence was obtained that lends support for preferential reduction of tetrazolium salts (e.g., nitroblue tetrazolium chloride) to that of O2 by the micelle-bound MPH. PMID- 2558979 TI - Oxidative modification of lens crystallins by H2O2 and chelated iron. AB - Crystallins are the soluble structural proteins that constitute approximately 90% of the dry mass of the eye lens. The present study attempts to elucidate possible mechanisms whereby the H2O2 present in the eye could contribute to the oxidative modification of lens crystallins. The data indicate that exposure of solutions of crystallins to H2O2 and EDTA-chelated iron leads to covalent crosslinking of polypeptides, loss of intrinsic protein fluorescence, and the generation of a novel fluorophor emitting in the 420 nm range. These changes closely mimic oxidative modifications that occur in lens proteins in vivo. Exposure of the proteins to H2O2 in the absence of chelated iron failed to generate detectable levels of these modifications. These findings are contrasted with earlier studies of lenses in organ culture where H2O2 alone produced marked damage while the further addition of chelated iron protected the lenses from oxidation. PMID- 2558980 TI - Source of superoxide anion radical in aerobic mixtures consisting of NAD[P]H, 5 methylphenazinium methyl sulfate and nitroblue tetrazolium chloride. AB - The source of superoxide anion radical (O2-.) in aerobic mixtures consisting of NAD[P]H, 5-methylphenazinium methyl sulfate (or its 1-methoxy derivative) and tetrazolium salt was investigated using superoxide dismutase (SOD), Mn(II), ferricytochrome-C, and epinephrine as probes. NAD[P]H + phenazine + O2 was found to reduce nitroblue tetrazolium, iodonitrotetrazolium, and thiazolyl blue in a manner sensitive to agents that dismutase O2-., viz., SOD and Mn(II). It also mediated the reduction of ferricytochrome-C, and augmented the autooxidation of epinephrine to the adrenochrome, without a tetrazolium salt present in the medium. The autooxidation of epinephrine, but not the reduction of ferricytochrome-C, was found to be sensitive to SOD. Nitroblue tetrazolium, either singly or in combination with SOD, did not stimulate the reduction of ferricytochrome-C. The oxidation of NADH, mediated by a catalytically low concentration of phenazine(+O2), was augmented two-fold by SOD. These observations are consistent with, and lend support to, a scheme of redox events (Scheme-3) wherein it is proposed that the source of O2-. in the NAD[P]H + phenazine + tetrazolium(+O2) system is the reduced phenazine, that the tetrazoinyl radical (a one-electron reduction product of tetrazolium) may not reduce O2 to O2-., that the redox reaction between semiquinone radicals of phenazine and O2 is reversible, and that the disproportionation of semiquinone radicals constitutes an important rate-limiting reaction in the expression of phenazine redox couple. PMID- 2558981 TI - Macroxyproteinase (M.O.P.): a 670 kDa proteinase complex that degrades oxidatively denatured proteins in red blood cells. AB - Erythrocytes and reticulocytes are shown to undergo rapid rates of protein degradation following exposure to oxidative stress. Experiments with ATP depletion revealed that, unlike the proteolysis of many other abnormal proteins, the degradation of oxidatively modified proteins is an ATP-independent process. Ion exchange chromatography (DEAE Sepharose CL-6B), ammonium sulfate precipitation, gel filtration chromatography (Sephacryl S-300 or Sepharose CL 6B), and a second ion exchange step were used to resolve the activity responsible for degrading oxidatively modified proteins from (dialyzed) cell-free extracts of erythrocytes and reticulocytes. Gel filtration studies revealed that some 70-80% of the activity in erythrocytes, and some 60-70% of the activity in reticulocytes, is expressed by a 670 kDa proteinase complex that is not stimulated by ATP (in fact, ATP is slightly inhibitory). This proteinase complex is inhibited by sulfhydryl reagents, serine reagents, and transition metal chelators, and has a pH optimum of 7.8. We propose the trivial name "macroxyproteinase" or "M.O.P." (abbreviated from Macro-Oxy-Proteinase) for the complex because of its large size, substrate preference (oxidatively modified proteins), and inhibitor profile (which indicates multiple catalytic sites). Electrophoresis studies of the 670 kDa M.O.P. complex revealed the presence of 8 distinct polypeptide subunits with the following apparent molecular sizes: 21.5, 25.3, 26.2, 28.1, 30.0, 31.9, 33.3, and 35.7 kDa. The large molecular size of the M.O.P. complex, its ATP- and ubiquitin-independence, its inhibitor profile, its distinctive subunit banding pattern in denaturing electrophoresis gels, its pH optimum, and its proteolytic profile with fluorogenic peptide substrates all indicate that M.O.P. is identical to 600-700 kDa neutral/alkaline proteinase complexes that have been isolated from a wide variety of eucaryotic cells and tissues, but for which no function has previously been clear. We propose that macroxyproteinase is responsible for catalyzing most of the selective degradation of oxidatively denatured proteins in red blood cells. We further suggest that M.O.P. may perform the same function in other eucaryotic cells and tissues. PMID- 2558982 TI - Papers presented at the Research Society on Alcoholism Symposium. June 3, 1988, Wild Dunes, S.C. PMID- 2558983 TI - Lipid peroxidation, iron mobilization and radical generation induced by alcohol. AB - Increasing evidence points to a major role for free radicals in the pathogenesis of alcohol-induced liver injury. In vitro, free radicals may be generated during ethanol metabolism by the further metabolism of acetaldehyde by molybdenum dependent oxidases such as xanthine oxidase. Ferritin iron mobilized by such free radicals may serve as catalytic iron. Increased stores of ferritin iron and induction of microsomal P-450 reductase activity are mechanisms by which chronic alcohol feeding may potentiate the acute effects of alcohol. PMID- 2558984 TI - Oxygen radical generation by microsomes: role of iron and implications for alcohol metabolism and toxicity. AB - Experiments were carried out to evaluate whether the molecular mechanism for ethanol oxidation by microsomes, a minor pathway of alcohol metabolism, involved generation of hydroxyl radical (.OH). Microsomes oxidized chemical .OH scavengers (KMB, DMSO, t-butyl alcohol, benzoate) by a reaction sensitive to catalase, but not SOD. Iron was required for microsomal .OH generation in view of the potent inhibition by desferrioxamine; however, the chelated form of iron was important. Microsomal .OH production was effectively stimulated by ferric EDTA or ferric DTPA, but poorly increased with ferric ATP, ferric citrate, or ferric ammonium sulfate. By contrast, the latter ferric complexes effectively increased microsomal chemiluminescence and lipid peroxidation, whereas ferric EDTA and ferric DTPA were inhibitory. Under conditions that minimize .OH production (absence of EDTA, iron) ethanol was oxidized by a cytochrome P-450-dependent process independent of reactive oxygen intermediates. Under conditions that promote microsomal .OH production, the oxidation of ethanol by .OH becomes more significant in contributing to the overall oxidation of ethanol by microsomes. Experiments with inhibitors and reconstituted systems containing P-450 and NADPH P-450 reductase indicated that the reductase is the critical enzyme locus for interacting with iron and catalyzing production of reactive oxygen species. Microsomes isolated from rats chronically fed ethanol catalyzed oxidation of .OH scavengers, light emission, and inactivation of added metabolic enzymes at elevated rates, and displayed an increase in ethanol oxidation by a .OH-dependent and a P-450-dependent pathway. It is possible that enhanced generation of reactive oxygen intermediates by microsomes may contribute to the hepatotoxic effects of ethanol. PMID- 2558985 TI - Duodenal mucosal bicarbonate secretion in man. Stimulation by acid and inhibition by the alpha 2-adrenoceptor agonist clonidine. AB - A multi-channel small diameter tube was used to study the secretion of bicarbonate by 3 cm long segments of the proximal duodenum isolated between balloons. The tube had an outer diameter of 5.3 mm and two central and four smaller, peripheral channels. Measurements of infused phenol red, 14C-PEG and vitamin B12 and of trypsin activity were performed to rule out contamination of the perfusate by gastric and pancreatic secretions. Basal secretion of bicarbonate by the duodenal mucosa in healthy subjects varied between 135 and 220 mumol/cm of intestine per hour. Perfusion of the lumen with acid (100 mM HCl for five minutes) increased the secretion to greater than 400 mumol/cm/h and the alpha 2-adrenoreceptor agonist clonidine (150 micrograms iv) decreased the HCO3- secretion by 70 mumol/cm/h. Clonidine simultaneously reduced the mean arterial blood pressure and plasma noradrenaline concentration, but did not affect the plasma glucose or adrenaline concentration. Duodenal bicarbonate secretion is important in the protection of this mucosa against acid discharged from the stomach. Increased sympathetic activity may, by inhibiting the bicarbonate secretion, decrease the protection in proximal duodenum in man and facilitate ulceration. PMID- 2558987 TI - [Analysis of PAP III D findings as a discussion principle for clinical consequences]. PMID- 2558986 TI - [Cytologic signs of human papillomavirus infection in HIV-positive patients]. PMID- 2558988 TI - [Non-immune hydrops fetalis caused by parvovirus infection: diagnosis and successful treatment]. PMID- 2558989 TI - [Micrometastases in axillary lymph nodes in breast cancer]. PMID- 2558990 TI - [The value of amniofetography for prenatal diagnosis]. PMID- 2558991 TI - [Treatment of in situ tubal pregnancy with prostaglandins]. PMID- 2558992 TI - [Cell receptors and proliferation markers in breast cancer]. PMID- 2558993 TI - [Detection of papillomavirus types 6/11, 16/18 and 31/33/35 using DNA/RNA hybridization. Initial experiences]. PMID- 2558994 TI - [Clinical significance of human papillomavirus infection]. PMID- 2558995 TI - [The value of cervix curettage in dysplasia of the cervix]. PMID- 2558996 TI - [Increased incidence and value of cytologic findings PAP III D]. PMID- 2558997 TI - [Clinical and histologic findings in persistent PAP III D]. PMID- 2558998 TI - [The incidence of papillomavirus types 6/11 and 16/18 in cervix smears]. PMID- 2559000 TI - [Follow-up of papillomavirus-induced changes of the cervix in pregnancy]. PMID- 2558999 TI - [Human papillomavirus infection as an oncogenic factor in the pathogenesis of cervix cancer]. PMID- 2559001 TI - [Microbiologic findings including Chlamydia serology in cervix dysplasia and human papillomavirus infection]. PMID- 2559002 TI - [The significance of multiple bacterial and viral infections in the origin of cervix dysplasia]. PMID- 2559003 TI - [The treatment of virus-associated cervix dysplasia with recombinant interferon alpha-2c or gamma]. PMID- 2559004 TI - [Laboratory diagnosis of Entameba histolytica]. PMID- 2559005 TI - [Antepartum prophylaxis: the final step in the Rh isoimmunization problem?]. PMID- 2559006 TI - [Detection of antigen and antibody in herpes simplex encephalitis]. AB - Laboratory confirmation of herpes simplex (HSV) infection in patients suspected of HSV encephalitis (HSV-E) at the earliest stage of the disease, may contribute greatly to the differential diagnosis and to the initiation of effective antiviral treatment. Our diagnosis of HSV infection was based on: a) detection of viral antigen in CSF cells in the first week of disease by immunofluorescence assay employing monoclonal antibodies against HSV-1 or HSV-2; b) detection of local IgM- and IgG-specific antibodies to HSV in the CSF and in the serum; c) ratio of titers of HSV antibodies in CSF and in serum less than 1:20; d) 4-fold rise in antibody titer to HSV in CSF and/or serum. The incidence of HSV-E was examined through 1987 in 270 patients suspected of viral infection of the central nervous system. In 187 (69.5%) material for laboratory diagnosis was inadequate and in 75 (27.8%) no evidence for infection by HSV was found. HSV infection was confirmed in 8 (2.9%), comprising 9.6% of those with adequate material for laboratory diagnosis. In no case was HSV isolated from the CSF. The importance of adequate material for viral diagnosis by the laboratory is emphasized. PMID- 2559008 TI - [[Klippel-Trenaunay syndrome--an embryonic developmental disorder. A case report with reference to the orthodontic symptoms]. AB - The Klippel-Trenaunay syndrome is a disturbance in the development of the mesodermal and ectodermal tissues occurring in utero. It is characterized by the clinical triad of unilateral soft tissue and bone hypertrophy, varicosities and hemangiomas. For dentists the orofacial manifestations of the syndrome are interesting: unilateral hypertrophy of the maxilla and mandible, mandibular joint and alveolar processes and tooth germs. In addition, the gingival margins are thickened, eruption is premature and there is marked facial asymmetry. PMID- 2559007 TI - Drug identification problems in two suicides with neuromuscular blocking agents. AB - Cases of suicide committed with neuromuscular blocking agents have been investigated. Both cases involved anesthesiologists who took the muscle relaxants suxamethonium chloride (Sukolin) or pipecuronium bromide (Arduan) and a rapid acting barbiturate, 5-sec-butyl-5-ethyl-2-thiobarbituric acid (Inactin). The examinations were performed by using gas chromatography and mass spectrometry. The quantitative data obtained showed that doses useful in general anesthesiological practice were applied. Metabolites were found despite their fast metabolizing character. PMID- 2559009 TI - [Adrenoleukodystrophy and adrenomyeloneuropathy--clinical spectrum, pathobiochemical aspects, diagnosis and therapy]. AB - Adrenoleukodystrophy (ALD) is characterized by adrenal cortical insufficiency and progressive demyelination of the central nervous system with lethal outcome in childhood. Adrenomyeloneuropathy (AMN) represents a clinical variant of ALD with later manifestation during adolescence. The neurological symptoms of this form are slowly progressive spastic paraparesis and peripheral neuropathy. Other intermediate forms, which might be classified as adrenoleukomyeloneuropathy (ALMN), possibly represent an overlap of the two conditions. Heterozygote carriers of the X-linked disease may also show neurological symptoms, such as spastic paraparesis and peripheral neuropathy. Biochemically, all these X-linked forms of ALD/AMN are characterized by an accumulation of very-long-chain fatty acids in various tissues and body fluids indicating an impaired peroxisomal metabolism. In this study two families with ALD are presented. The two forms of ALD and AMN were observed in different members of the families including a heterozygote female carrier with neurological symptoms. The different clinical syndromes, the biochemical and genetic basis, and new therapeutical strategies are discussed. PMID- 2559010 TI - [Mucin-producing signet ring cell carcinoma of stomach accompanied by microangiopathic hemolytic anemia and disseminated intravascular coagulation: a case report]. AB - Diffuse bone marrow metastasis of carcinoma sometimes accompanies microangiopathic hemolytic anemia (MHA) which is characteristic with red cell fragmentation. We report on a 56-year-old male case of signet ring cell type gastric carcinoma complicated with MHA and DIC. Bone marrow aspiration disclosed the metastasis of carcinoma cells. However, no other distant metastasis was evident by pathological studies of autopsy samples. PMID- 2559011 TI - Activators and inactivators of calcium channels: effects in the central nervous system. AB - The interactions of calcium antagonists or channel activators with the different classes of calcium channel are reviewed with particular emphasis on interactions with neuronal tissue; reasons for the failure of calcium antagonists to inhibit neurotransmitter release under normal circumstances are outlined. Calcium antagonists may be protective in several pathological situations and the possibilities of protection against ischaemic damage in the central nervous system are evaluated. PMID- 2559012 TI - Central neurochemical effects of dihydropyridine calcium antagonists. AB - Recent advances in central dihydropyridine (DHP)-binding sites are reviewed. DHP binding sites are pre-synaptically and post-synaptically localized in the brain. The functional role of post-synaptic sites is still unknown, whereas pre-synaptic sites seem to contribute to the control of calcium uptake and of neurotransmitter release. DHP-binding sites may be modualated in physiological (age, sex) and pathological events (hypertension, ischaemia, neurological diseases) or after drug treatments (alcohol, morphine, etc.). The reviewed data suggest new therapeutic implications of DHP calcium channel antagonists in the treatment of other diseases and of drug withdrawal syndrome. PMID- 2559013 TI - Decreased melatonin concentration in Cushing's syndrome. AB - To determine the effect of hypercortisolaemia on the melatonin circadian secretion 12 patients with pituitary or adrenal dependent Cushing's syndrome and 5 healthy controls were studied. The melatonin circadian rhythm of secretion, observed in the control group, was abolished in the patients with hypercortisolaemia. Mean nocturnal melatonin levels and the integrated 24-hour secretion were significantly lower in the patients studied than those of the controls. Thus, in patients with Cushing's syndrome the melatonin levels are decreased and the circadian rhythm of this hormone is abolished. PMID- 2559014 TI - Hyperprolactinemia in hepatic encephalopathy may result from impaired central dopaminergic neurotransmission. AB - Ten patients with liver disease and hepatic encephalopathy (HE) and eight normal controls were studied. Five of the 10 HE patients had hyperprolactinemia. The administration of L-dopa produced a decrease of serum prolactin in all. Prior administration of Carbidopa, a peripheral decarboxylase inhibitor, did not change the prolactin suppression by L-dopa in the normal controls or in the patients with normal baseline prolactin levels. In the hyperprolactinemic group, Carbidopa significantly inhibited the response to L-dopa. Impaired central neurotransmission, at least involving the hypothalamic-pituitary dopaminergic system, may underlie the hyperprolactinemia in HE. PMID- 2559015 TI - Isolation of IgG islet cell autoantibody-producing B lymphocytes from the peripheral blood of type 1 diabetic patients and an ICA-positive non-diabetic individual. PMID- 2559017 TI - Possibilities and limits of the medical treatment for primary hyperparathyroidism. AB - Clinically obvious primary hyperparathyroidism is only curable by surgery. Medical treatment is debatable under other circumstances: mild chronic hypercalcemia, patients who refuse surgery, serious coexisting medical problems and recurrence or persistence of PHPT after surgical treatment. Prior to medical treatment, the usual common management of any mild hypercalcemia must be taken. The potential medical treatments are: (1) the inhibition of parathyroid hormone (PTH) secretion, and (2) the inhibition of the effects of PTH. The substances of these two main types are successively described. Nevertheless, no ideal medical treatment of PHPT is actually available. PMID- 2559016 TI - Effects of short-term glucocorticoid deprivation on growth hormone (GH) response to GH-releasing hormone in man. PMID- 2559018 TI - Ability of cofactors to support peptide amidation is cell-type specific. AB - The ability of various cofactors to substitute for ascorbate in the biosynthesis of alpha-aidated peptides from pro-ACTH/endorphin (PAE) was compared in corticotrope tumor cells (AtT-20) and in primary anterior and intermediate pituitary cultures. In all three systems, ascorbate was the most potent cofactor tested. In AtT-20 cells, dopamine, norepinephrine, epinephrine, dehydroascorbate and dihydro- and tetrahydrobiopterin supported significant alpha-amidation of joining peptide [PAE(77-94)NH2]. In contrast, amidation of joining peptide by primary corticotropes was stimulated only slightly by catecholamines and not by tetrahydrobiopterin. Neither catecholamines nor tetrahydrobiopterin stimulated peptide amidation by melanotropes. The ability of cofactors to support the synthesis of alpha-amidated peptides is cell-type specific. PMID- 2559019 TI - Factors that determine cell-specific gene expression in pancreatic endocrine tumor cells. AB - Phenotypically distinct islet tumor cell lines may recapitulate certain of the developmental pathways of normal islet cell differentiation by expressing a combinatorial set of positively and negatively acting DNA-binding proteins to allow for the programmed expression of genes encoding polypeptide hormones. The structure of one of these DNA-binding proteins, a cyclic AMP-responsive protein (CREB) that binds specific DNA regulatory elements in the somatostatin gene, has been deduced from the sequence of a cloned cDNA. The CREB protein contains a DNA binding domain separate from a cAMP-dependent protein kinase A activation domain. Further characterizations of the genes encoding the DNA-binding proteins should help to elucidate the cellular processes involved in islet cell differentiation and the genesis of tumors. PMID- 2559020 TI - Long-acting somatostatin analogues in pancreatic islet cell carcinoma. AB - In secreting islet cell carcinoma, the long-acting somatostatin analogue, Sandostatin, reduces symptoms of endocrine secretion both by inhibiting peptide secretion and by acting on the target organs. It could be used during the initial evaluation of patients with such tumors and thereafter when the tumor cannot be found or is metastatic. Its efficacy depends upon the tumor type and probably the presence of somatostatin receptors on the tumoral cells. It could decrease with time, especially in VIPomas. Side effects are few and usually mild. Hypoglycemia attacks in insulinoma could be worsened during treatment by the complete inhibition of hyperglycemic hormones. The inhibition of tumoral growth, based on animal models, appears infrequent in clinical practice. PMID- 2559021 TI - Pancreatic islet cell tumors metastatic to the liver: treatment by hepatic artery chemo-embolization. AB - The embolization of hepatic metastases of pancreatic islet cell tumors achieves a tumor necrosis without liver failure owing to double hepatic blood supply. The arterial chemotherapy performed at the same time delivers a large amount of cytotoxic agent directly into the tumor. Tumor bulk reducing and decreasing of production of pharmacologically active substances may be obtained to some degree. Although an objective documentation of the therapeutic effect is difficult to obtain, several series emphasize that the embolization provides a partial and transient palliative remission. Therefore, the chemo-embolization should be included in a multidisciplinary approach of metastatic endocrine malignancies in combination with surgical resection, systemic antineoplastic chemotherapy, and antihormonal therapy. PMID- 2559022 TI - Detection of human papillomavirus DNA in semen from patients with intrameatal penile warts. AB - Fifteen semen specimens from 10 men with intrameatal penile warts attending a genitourinary clinic were tested by Southern blot hybridisation for the presence of human papillomavirus (HPV) DNA. Five specimens were positive for HPV types 6/11. This observation may have implications for screening of semen used for artificial insemination by donor. PMID- 2559023 TI - Acquisition and retention of viruses by Trichomonas vaginalis. AB - Recently described occurrence of virus-like particles (VLP) in some strains of Trichomonas vaginalis suggests the possibility that the pathogenic significance of this organism may be broadened by its potential for viral transmission. Inasmuch as neither the source nor the host range of the VLP are known, any hazard which they may present for man cannot be estimated. A model has been established for the study of acquisition of known human viruses by T vaginalis. Tissue cultures were infected with two reoviruses and a fresh isolate of genital herpes simplex virus (HSV). A squirrel monkey reovirus was also included in the study. T vaginalis was inoculated into the virus cultures three days later. The progress of virus acquisition by the trichomonads was monitored by transmission electron microscopy and by culture. Virus-containing cell fragments were engulfed by trichomonads and internalised in vacuoles. After digestion of cellular debris only virus particle aggregates were retained. Viable reoviruses were recovered from the trichomonads for nine days, and HSV for six days. The results suggest the possibility of transmission of at least some viruses by T vaginalis. PMID- 2559024 TI - Characterization of two genes for the human Na,K-ATPase beta subunit. AB - A total of 29 human genomic DNA clones that hybridize with cDNAs for the sheep and rat Na,K-ATPase beta subunits have been isolated, classified by restriction endonuclease mapping and Southern blot hybridization analysis, and sequenced. One class of clones, designated ATP1BL1, represents a processed pseudogene for the beta subunit. The second class, designated ATP1B, includes 15 overlapping genomic clones and represents a functional gene for the human Na,K-ATPase beta subunit. ATP1B spans about 26.7 kb of genomic DNA and includes 24 kb of intron sequence. The complete mRNA transcript for the human beta subunit is encoded by six exons, ranging in size from 81 to 1427 bp. Primer extension and S1 nuclease protection experiments with human kidney RNA indicate the presence of two major transcription initiation sites at -510 and -201 to -191, with minor initiation sites at -268, -182 to -174, and -142. The distal initiation site at -510 is preceded by consensus sequences for CAAT and TATA boxes. The DNA sequence preceding the proximal heterogeneous initiation sites contains a CAAT box, but no TATA box. Two of the 12 GC boxes (GGCGGG and CCCGCC) located in the 5' region of ATP1B are located between this CAAT box and the proximal clusters of transcription initiation sites. PMID- 2559025 TI - Regional localization of the human thrombomodulin gene to 20p12-cen. PMID- 2559026 TI - [Genetic regulatory mechanisms of bacterial virulence]. AB - Bacterial virulence is due to the action of different virulence factors (e.g. adhesins, toxins, capsules). Virulence factors are encoded by special genes termed as virulence genes or virulence determinants. The degree of virulence of one particular bacterial strain depends on the activity of the corresponding virulence genes. Several mechanisms are involved in the regulation of activity of virulence determinants: plasmids and bacteriophages may be received or lost and chromosomal sequences may be deleted. Such processes are directly responsible for the presence or absence of virulence genes in the bacterial genome. Programmed genetic rearrangements may lead to a switch of promoter sequences and are therefore responsible for a variation of virulence expression. "Jumping" of DNA sequences (transposition) and subsequent recombinational events may also cause antigenic variation of virulence factors. In addition frame shift mutations may influence the expression of virulence genes. Transregulatory factors may also influence the expression of virulence factors of pathogenic bacteria. These systems positively influence the expression of different virulence factors in a coordinative manner. They can be triggered by environmental signals. PMID- 2559027 TI - Mechanisms of human neutrophil-mediated cartilage damage in vitro: the role of lysosomal enzymes, hydrogen peroxide and hypochlorous acid. AB - Cartilage is a focal point of attack by cellular and molecular elements of the inflammatory response which occurs in arthritic diseases. Neutrophils damage articular cartilage by degrading matrix components and inhibiting their synthesis. The aim of this study was to elucidate mechanisms of this damage. Human neutrophils were isolated from blood by centrifuging through Ficoll-Hypaque and granule extract prepared from them. Articular cartilage from adult humans and cattle was maintained in organ culture. Cartilage degradation (release of 35S labelled proteoglycan) or synthesis (incorporation of 35S into proteoglycan) was determined after various treatments. Human neutrophils and neutrophil granule extract degraded proteoglycan and inhibited proteoglycan synthesis. The specific leucocyte elastase inhibitor N-methoxysuccinyl-(ala)2-pro-val-chloromethylketone (MAAPVCMK) partially reversed these effects. H2O2, a product of the neutrophil respiratory burst, when added directly at 10(-6)mol/L, or generated by glucose oxidase (GO)/glucose inhibited proteoglycan synthesis but had no effect on degradation. Hypochlorous acid (OHCl), a product of the myeloperoxidase (MPO)/H2O2/Cl system at 50 mumol/L degraded proteoglycan and inhibited its synthesis. OHCl produced by granule extract (as a source of MPO) + GO-generated H2O2 + Cl- degraded proteoglycan. The results indicate that neutrophil-mediated proteoglycan degradation and inhibition of synthesis is largely attributable to elastase and secondarily to OHCl, whereas H2O2 impairs synthesis without affecting degradation of proteoglycan. PMID- 2559028 TI - [Differential diagnosis of multiple gliomas, multicentric gliomas and the other main endocranial multifocal lesions. Presentation of a case]. AB - Multifocal gliomas represent an uncommon lesion, particularly if the supra tentorial and infra-tentorial spaces are involved. The Authors describe a particular case and they outline the main differences between multiple gliomas and multicentric gliomas. Then, they consider the various possibilities of diagnosis and differential diagnosis of multiple endocranial lesions. PMID- 2559029 TI - Purification of phosphatidylinositol-specific phospholipase C from NG108-15 cells. AB - The phosphatidylinositol-specific phospholipase C from the membrane of NG108-15 cells has been purified to homogeneity by using DEAE Bio-Gel A agarose, hydroxyapatite, and heparin agarose chromatography. The purified phosphatidylinositol-specific phospholipase C has been purified 422-fold and its molecular weight has been estimated to be 28,000. We have obtained a final specific activity of 3.8 mumols of phosphatidylinositol hydrolyzed/min/mg of protein. The purified enzyme is specific to phosphatidylinositol. The purified enzyme has an apparent pH optimum of 7.0. Calcium ions are required for its activity. PMID- 2559030 TI - Alterations in complement-induced shape change and stimulus-specific superoxide anion generation by neonatal calf neutrophils. AB - Increased susceptibility of neonates to infection may be related to defects in newborn neutrophil (PMN) functional activities, including altered responses to complement fragments (Cf) and defective microbicidal activity. We therefore compared the kinetics of newborn and adult bovine PMN membrane shape change responses following stimulation with zymosan-activated plasma (ZAP) as a source of Cf. Measurement of PMN membrane shape change was a rapid, sensitive, and reproducible measure of Cf stimulation within a population of PMNs; a maximum of 67-85% of the PMNs exhibited easily detectable membrane ruffling, lamellipodia formation, and polarity within 2 min. Newborn PMNs exhibited significantly increased (P less than 0.01) membrane shape change at 20, 30, 60, 120, and 300 sec after Cf stimulation. A maximum of 85.8 +/- 3.2% of newborn PMNs exhibited such Cf-induced shape changes by 120 sec. which was significantly greater (P less than 0.01) than the maximum stimulation (67.7 +/- 4.3%) attained with adult PMNs. These data indicate enhanced kinetics of induced newborn PMN membrane shape change in response to Cf stimulation. We also compared stimulus-specific superoxide anion (O2-) generation as a measure of respiratory burst activity after incubation of newborn and adult PMNs with soluble (phorbol myristate acetate, PMA) and particulate (opsonized zymosan, OZ) stimuli. When PMA was used as the stimulus, newborn PMNs generated significantly less O2- (9.3 +/- 0.5 nmol O2-/10(6) PMN, P less than 0.05) than did adult PMNs (12.4 +/- 0.3 nmol O2-/10(6) PMN). This finding was reversed when OZ was used as the stimulus; newborn PMNs generated significantly more O2- (7.7 +/- 0.4 nmol O2-/10(6) PMN, P less than 0.05) than did adult PMNs (5.5 +/- 0.5 nmol O2-/10(6) PMN). These findings collectively document biochemical and morphological differences between newborn and adult PMNs as determined by stimulus-specific O2- generation and Cf-induced membrane shape change. Such differences may be important to neonatal disease susceptibility. PMID- 2559031 TI - Inhibition by halothane, but not by isoflurane, of oxidative response to opsonized zymosan in whole blood. AB - In an attempt to clarify some apparent discrepancies in reports of the effects of anesthetic agents upon granulocyte function, we studied the effects of halothane and isoflurane, using techniques that allowed us to perform the assays in and in sealed vials to prevent volatile gas evolution: assay gas concentrations were measured, rather than inferred. Chemiluminescence, superoxide production, and hydrogen peroxide production were assessed after presentation of opsonized zymosan as a phagocytic stimulus. Incubation with halothane led to a highly statistically significant dose-related inhibition of chemiluminescence (maximum 66%), H2O2 production (67%) and .O2- production (61%), within the concentration range observed in blood from patients undergoing general anesthesia. In contrast, the presence of isoflurane led to no statistically significant changes in any of the functions measured. Cells harvested from patients undergoing elective halothane anesthesia showed the same functional inhibition, but for quantitative differences likely due to the inability to control for dilution effects in clinical samples. It has been suggested that halothane anesthesia may be associated with excess mortality in septic patients; although the results we report are readily reversible, their presence during a prolonged anesthesia could be harmful in a patient who is not immunologically normal and/or who is already infected. Careful clinical trials will be necessary to determine if isoflurane is a superior agent in this context. PMID- 2559033 TI - Localization of islet cell tumors. AB - In summary, although similar imaging techniques are used for the localization of insulin and gastrin secreting islet cell tumors, the success rates are very different. Fortunately, the best results are obtained in the tumor for which effective medical treatment is less available, namely insulinomas, which can be found and successfully resected in over 90 per cent of patients. Both portal venous sampling and experienced intraoperative ultrasound are critical for successful surgery of small insulin-secreting tumors. Facilities without these resources should not explore patients when the conventional imaging studies are negative. Gastrinomas, on the other hand, will elude detection by even the most experienced surgeons in over 20 per cent of patients with sporadic Zollinger Ellison syndrome in spite of positive portal venous sampling and intra-arterial secretin studies. The very small size of these tumors and their occurrence in more difficult to explore extrapancreatic sites provides the basis for this difference. However, patients with negative imaging studies and negative surgical explorations have an excellent prognosis on long-term follow-up when gastric acid hypersecretion is controlled. An annual computed tomographic scan is recommended since the appearance of a tumor would mandate surgical resection because of the significant incidence of malignancy in larger tumors. However, progression to imageable tumors has been unusual in our experience with this group of patients. There remains a small group of patients with highly malignant, rapidly metastasizing, gastrin-secreting islet cell carcinomas for whom localization is simple but of little relevance. However, locally invasive gastrinomas may often be resectable and provide prolonged remission and even cure. Aggressive surgery, supported by detailed cross-sectional and angiographic localization, has a role in this small group of patients. PMID- 2559034 TI - The role of surgery in the management of islet cell tumors. AB - Endocrine tumors of the pancreas are rare disorders that can cause life threatening symptoms related to the excessive hormonal secretion and the malignant nature of the neoplasm. In addition, these neoplasms can present as part of familial endocrinopathy syndromes, especially multiple endocrine neoplasia type 1 (MEN-1). The initial step in evaluation of a patient with an islet cell tumor is definitive diagnosis of the hormonal syndrome that usually can be achieved biochemically. The next step is to reverse the life-threatening hormonal syndrome medically to allow radiographic localization of the islet cell neoplasm and to prepare the patient for elective surgery. In general, the goal of surgery is to accurately stage the extent of disease and to totally resect the tumor resulting in complete correction of the excessive hormonal condition and freedom from malignant progression of the tumor. Resection of the neoplasm should encompass metastatic disease in select individuals in whom complete or nearly complete resection can be achieved. Surgery must be performed with acceptable morbidity and mortality because the medical management of the excessive hormonal secretion in most patients is adequate and the progression rate of malignant islet cell carcinomas is usually slow. Patients with MEN-1 should be identified during evaluation by careful history and should be managed differently. These patients always have multiple islet cell tumors and may have malignant islet cell carcinomas. Large (3 cm) imageable pancreatic neoplasms in patients with MEN-1 should be resected because nearly 50 per cent are malignant. Zollinger-Ellison syndrome in patients with MEN-1 and primary hyperparathyroidism is best managed initially by surgery directed at the hyperparathyroidism. It may be impossible to correct the biochemical abnormalities of Zollinger-Ellison syndrome by islet cell tumor resection in patients with MEN-1. However, insulinoma or VIPoma syndromes can often be corrected by islet cell tumor resection. Portal venous sampling for hormones may be helpful in determining what hormone a specific tumor is secreting. PMID- 2559035 TI - Glucagonomas and insulinomas. AB - The clinical features, diagnosis, differential diagnosis, and treatment of the glucagonoma and insulinoma syndromes as well as the prevalence, distribution, malignancy, and localization of the two tumors are described. PMID- 2559032 TI - Inhibition of inflammatory cell infiltration by bicyclic imidazoles, SK&F 86002 and SK&F 104493. AB - The mode of action of the dual inhibitors of eicosanoid metabolism, SK&F 86002 and SK&F 104493 was evaluated on inflammatory cell infiltration induced in mice by carrageenan, monosodium urate crystals, and arachidonic acid. The results were compared to those seen with standard antiinflammatory compounds. Inflammatory cell infiltration was inhibited by SK&F 86002. SK&F 104493, colchicine, and phenidone but not naproxen. In vivo, PMN infiltration induced by LTB4 was inhibited by colchicine but not by SK&F 86002, SK&F 104493, or phenidone treatment. Similarly, in vitro chemotaxis to LTB4 was not inhibited by SK&F 86002. The 5-lipoxygenase inhibitors, SK&F 86002, SK&F 104493, and phenidone inhibited LTB4 production in vivo as well as inflammatory cell infiltration induced by arachidonic acid. The data are consistent with the suggestion that the bicyclic imidazoles inhibit PMN infiltration by virtue of inhibition of LTB4 production. PMID- 2559036 TI - The use of the long-acting somatostatin analogue, octreotide acetate, in patients with islet cell tumors. AB - Octreotide lowers plasma concentrations of the marker peptide in the majority of patients with islet cell tumors. However, as described above the effect of octreotide on plasma concentrations of marker peptides is not necessarily related to the effect on symptoms. Nevertheless octreotide is capable of producing symptomatic relief in a large proportion of patients with islet cell tumor syndromes. The data on the effect of octreotide on the symptoms due to VIPoma and due to the carcinoid syndrome (presumably including some who have islet cell tumors) are strong and the drug has been approved for these indications by the Food and Drug Administration. With respect to the other islet cell tumor syndromes, the published data suggest that the utility of octreotide differs in the different syndromes. Insulinomas are usually single, benign, and can and should be removed surgically, resulting in cure. Octreotide therefore has no role to play in such patients, particularly since the response of insulinomas is variable. However in the 10 per cent of insulinomas that are malignant octreotide is certainly effective in at least a portion of cases, although as yet the true response rate and efficacy compared with diazoxide is not clear. Although octreotide is effective at reducing acid output, and thus improving symptoms in patients with Zollinger-Ellison syndrome, because of the effectiveness of histamine H2-receptor antagonists and omeprazole, there is no need for octreotide in this syndrome. For patients with glucagonoma, GHRHoma, Cushing's syndrome, and other rare islet cell tumor syndromes octreotide may well be of benefit and should be considered. The current data do not support the use of octreotide for an antitumor effect. PMID- 2559037 TI - Systemic and regional therapy of advanced islet cell tumors. AB - Asymptomatic patients with islet cell tumors should be observed every 3 to 6 months. Once symptomatic, many therapeutic choices exist. First choice would be symptomatic management resulting in minimum side effects. More aggressive approaches include systemic therapy with cytotoxic or biologic agents. Vascular occlusion provides useful palliation of liver metastases. PMID- 2559039 TI - Phorbol myristate acetate stimulates formation of diphosphoinositide in rat mast cell granules. AB - Rat mast cell granules were obtained by homogenization of highly purified rat mast cells and isolated in a Percoll gradient. Diphosphoinositide (DPI) synthesis in rat mast cell granules was assayed by measuring the incorporation of 32P from [gamma 32P]ATP into DPI in the absence of exogenous phosphatidylinositol (PI). Lipids were isolated with methanol/chloroform/HCl and were separated by thin layer chromatography on oxalic acid impregnated silica gel plates. DPI areas were identified by staining with iodine, scraped and measured for 32P radioactivity. The addition of phorbol myristate acetate (PMA) to the granules caused an increase of 32P incorporation from [gamma 32P]ATP in the DPI fraction, which can be catalyzed by PI kinase. This effect of PMA in the DPI synthesis was dose dependent and maximal effects were observed at 10 ng/ml. PMID- 2559038 TI - Influence of Ly146032 on chemotaxis, chemiluminescence of PMN and lymphocyte transformation in vitro. AB - The influence of Ly146032, a new acidic lipopeptide, on chemotaxis, luminol dependent chemiluminescence of human polymorphonuclear leukocytes (PMN) and phythemagglutinin induced lymphocyte transformation of murine cells was investigated. At therapeutic range there was no remarkable effect on the parameters tested. Incubation with more than 20 mg/l Ly146032 was followed by depression of chemiluminescence, whilst transformation of maximally PHA stimulated lymphocytes was suppressed by more than 32 mg/l Ly146032. PMID- 2559041 TI - Muramyl peptides confer hepatoprotection against murine viral hepatitis. AB - The hepatoprotection induced by synthetic muramyl peptides was investigated using a model of lethal murine mouse hepatitis MHV-3 virus infection. MDP and a nonpyrogenic analog, Murametide, inhibited the steep elevation of serum transaminases induced by MHV-3 irrespective of whether the immunomodulators were administered before or after the infection. A significant proportion of MDP or Murametide-treated animals, in contrast to controls, survived the MHV-3 infection. The histopathological examination of the liver revealed marked necrosis of the hepatic parenchymal cells and infiltration of the inflammatory cells in controls but not in MDP-treated animals. PMID- 2559040 TI - Effects of oral base therapy on serum ionized calcium, phosphorus and parathyroid hormone in chronic hemodialysis patients. AB - The purpose of this study was to evaluate the effects of oral base therapy on selected chemical parameters in chronic hemodialysis patients. Oral base supplements were administered to 20 acidotic chronic hemodialysis patients for one month. Serum bicarbonate levels rose from 18.6 +/- 2.9 to 22.5 +/- 4.0 mEq/L (p less than 0.0005) and pH rose from 7.35 +/- 0.03 to 7.39 +/- 0.04 (p less than 0.0005). Serum ionized calcium levels fell from 5.03 +/- 0.37 to 4.83 +/- 0.34 mg/dL (1.25 +/- 0.09 to 1.21 +/- 0.08 mmol/L) (p less than 0.01), while intact parathyroid hormone (PTH) levels rose from 547 +/- 697 to 619 +/- 776 pg/mL (p less than 0.05). Base therapy did not result in significant changes in serum levels of total calcium, phosphorus, alkaline phosphatase, urea nitrogen, creatinine, total protein, albumin or potassium. If empiric therapy with exogenous base is given to dialysis patients, ionized calcium levels should be closely monitored since changes in calcium supplement or vitamin D therapy may be required to maintain ionized calcium and parathyroid hormone values at the pre treatment levels. PMID- 2559042 TI - Comparative effect of the naphthalenic ansamycins rifamycin SV, rifampin and cyclopentylrifampicin on murine neutrophil function. AB - The purpose of this study was to evaluate the in vitro effect of the naphthalenic ansamycins rifamycin SV, rifampin and cyclopentylrifampicin on neutrophil degranulation and cytokinesis using murine peritoneal exudate cells. It was found that the FMLP-stimulated secretion of myeloperoxidase was significantly inhibited by 80 micrograms rifamycin SV (P less than 0.05) and cyclopentylrifampicin (P less than 0.01) per ml. Nondirected and FMLP-directed migration was significantly (P less than 0.01) inhibited by rifamycin SV and rifampin at assay concentrations above 0.31 and 5.0 micrograms/ml respectively thus confirming the low dose rifamycin effect observed by others using human neutrophils. Finally, cyclopentylrifampicin was shown to have no significant effect on nondirected or FMLP-directed neutrophil migration at assay concentrations of 1.25 to 80 micrograms per ml. PMID- 2559043 TI - Sympathetic control of specific and non-specific immune response. AB - Following antigenic stimulation with specific (alloantigen) and non-specific (sheep red blood cells (SRBC) and lipopolysaccharide (LPS] antigens the expression of beta-adrenergic receptors and the interference of immune sera IgG, on uterine smooth muscle beta-adrenoceptor-specific ligands were studied. The binding of alloimmune IgG to beta-adrenoceptors appeared to be specific, because immunization with SRBC and LPS did not induce anti-beta-adrenoceptors antibodies. On the contrary, the decrease in beta-adrenergic receptor expression was observed even with alloantigens and with the conventional antigenic challenge, suggesting that this phenomenon could be a non-specific immunoregulatory mechanism. PMID- 2559044 TI - VLCD compliance and lean body mass. AB - Very low calorie diets (VLCD) have many advantages, as they are inexpensive, safe and easy to comply with, and give rapid and encouraging weight loss. On the other hand, many patients complain of hunger and constipation. We have shown that these drawbacks can be reduced by the supplementation of 30 g of dietary fibre to VLCD. By the use of a VLCD which provides approximately 60 g of protein for women and approximately 70 g for men, the dietary regimen is safe and no excessive loss of lean body mass seems to occur during VLCD in obese patients. However, the changes in body composition that may occur after cycles of weight loss and regain have not been clarified. After weight cycles with weight losses obtained by conventional diets, obese women have lower lean body mass than obese non-dieting controls. Hence, more information about the changes in body composition during dieting on conventional diets and VLCD are needed. PMID- 2559045 TI - Cellular energy status of the gastric mucosa and gastric mucosal prevention by vitamin A in indomethacin-treated rats. AB - To evaluate the cellular energy status of gastric mucosa during the development of gastric mucosal damage and its prevention by application of a cytoprotective scavenger (vitamin A) in rats treated with indomethacin, the tissue levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP) and lactate were measured enzymatically, while the concentration of cyclic adenosine monophosphate (cAMP) was measured by radioimmunoassay, 4 h after the treatment with indomethacin and vitamin A simultaneously. It was found that a) the tissue levels of ATP, cAMP, AMP and the ratio of ATP/ADP were increased dose-dependently in the gastric mucosa in connection with the development of gastric mucosal prevention produced by vitamin A, b) the tissue level of ADP was decreased dose-dependently by vitamin A in indomethacin-treated rats, and c) the value of "energy charge" and the ratio of the ATP-ADP was unchanged during the vitamin A treatment in indomethacin-treated rats. The presence of tissue hypoxia could not be proved either in the development of indomethacin-induced gastric mucosal damage nor in the prevention of gastric mucosal damage by vitamin A. PMID- 2559046 TI - Rapid enhancement of cAMP accumulation in rat brain particulate fraction after ischaemia. AB - Ischaemia rapidly increases the concentration of cAMP in the brain by mechanism(s) which still remain undefined. In the present study significant enhancement of cAMP generation was found in vitro after brain ischaemia induced by decapitation or cardiac arrest. The particulate fraction from ischaemic brain accumulated considerably more cAMP and responded more effectively to stimulation by noradrenaline (NE), histamine (Hi) and adenosine than that prepared from normoxic controls. The most pronounced effect was observed immediately after ischaemic insult and proceeded to normalize during 24 hours of postischaemic recovery. The activation of cAMP production by NE and Hi, but not 2 chloradenosine (2-CA), was totally dependent on the presence of endogenous adenosine and calcium. The synergism of 2-CA with NE or Hi on cAMP accumulation was observed. Its disappearance after adenosine deaminase treatment indicates that the postischaemic activation of adenosine A2 receptors may be positively modulated by NE and Hi. These results strongly suggest that primary functional changes in the cell-membrane signalling system had been induced by the ischaemic insult. PMID- 2559047 TI - High affinity binding sites for Perindopril a new inhibitor of angiotensin-I converting enzyme (ACE) in the rabbit kidney: possible evidence for localization of ACE in endothelial structures and in glomerular mesangium. AB - The tissue distribution of Perindopril, a new potent inhibitor of the angiotensin converting enzyme (ACE), was studied after in vivo intravenous injection into rabbits of tracer amounts of the tritiated drug either alone (no modification of the renin angiotensin system) or along with a pharmacologic dose of 10 mg/kg unlabelled Perindopril. Lung and kidneys were the most densely labelled tissues. Kidney distribution of tritiated Perindopril was studied either by histo autoradiography or by measurement of radioactivity in the homogenates of dissected kidney zones. A close parallelism was found between the distribution patterns of ACE and radioactivity throughout the kidney. Tritiated-Perindopril binding was inhibited by concurrent treatment of the animals with the unlabelled drug or pretreatment with Captopril. Autoradiographic study of kidney slices after the administration of tracer amounts of Perindopril showed an intense labelling of the glomerular mesangium and of endothelial structures of blood vessels, and a lack of labelling of tubular epithelial cells. The possible occurrence of two pools of "ACE-like" activity in the kidney is discussed, namely i) one pool which is labelled by tritiated Perindopril, located in glomerular mesangium and endothelial structures which may be involved in the pharmacological action of ACE inhibitors; and ii) a second pool located in the proximal-tubule cell brush-border, remaining unlabelled by Perindopril, for which the high amount of neutral endopeptidase present at this site may be responsible. PMID- 2559049 TI - Autoimmunity to receptors. PMID- 2559048 TI - The role of impairment of adrenal mineraloglucocorticoid function in the development of infertility in varicocele patients. AB - The study was aimed at the understanding of pathophysiological mechanisms of the impairment of spermatogenesis in varicocele patients. The crucial role of tension increase in the venous plexus of the spermatic cord in spermatogenesis damage in the testis on the varicocele side and absence of any effect of haemodynamic abnormalities on spermatogenesis in the complementary testis has been determined. Retrograde blood flow through the central vein of the left adrenal gland in varicocele has been evidenced by X-ray examination. The role of this phenomenon in the changes of functional activity of the adrenal gland is discussed. Results of the study of functional status of adrenal glands revealed their tendency to provide hyperactivity in synthesis of mineraloglucocorticoids. Correlation between cortisol level in peripheral blood and percentage of abnormal sperm in ejaculate was shown. This fact supported the idea about the existence of a causal interrelationship between abnormalities in the functional status of adrenal glands and development of infertility in varicocele patients. PMID- 2559050 TI - Atomic bomb and accident dosimetry with ESR: natural rocks and human tooth in vivo spectrometer. AB - ESR dosimetry of some construction materials at Hiroshima and Nagasaki was carried out to determine the A-bomb radiation dose. Some minerals exposed to low level natural radiation over a given geological time period can be also used to determine the intense A-bomb radiation dose. Finally, an ESR cavity and a special NdBFe (Neomax) magnet system for in-vivo measurement of radiation dose of a human tooth without extraction is designed and manufactured. PMID- 2559051 TI - Human tooth dosimetry for gamma-rays and dental x-rays using ESR. AB - The effect of secondary electron equilibrium on the ESR dosimetry of x- and gamma ray irradiation has been studied by using acrylic plates to simulate the soft tissue of the human mouth. Dental x-rays create a different ESR signal pattern at the front surface of the tooth vs the back surface. The effect on radiation build up of the acrylic plate on gamma-irradiation is dominant at the front but is not appreciable at the back side of the tooth. PMID- 2559052 TI - Dose estimation by ESR spectroscopy at a fatal radiation accident. AB - In Norway, 1982, a fatal radiation accident occurred at a radioactive cobalt source used for radiation processing. There was a lack of realistic estimates on the victim's tissue or body doses up to his death. GSF was able to provide the first reliable dose values, upon request from Norwegian colleagues. Dosimetry was performed by ESR spectroscopy of long-lived radiation-induced radicals that could be measured in the heart tablets of the victim. The concentration of the free radicals proved, by proper calibration, to be proportional to dose. Procedures and dose results are reported; comparison is made with data from literature dealing with the same subject and also using TL as well as ESR. For biological relevance, the doses as determined from the tablets are converted into whole-body doses. PMID- 2559053 TI - Mechanically-induced generation of radicals in tooth enamel. AB - Mechanical instrumentation of enamel leads to the formation of long-lived free radicals that can be conveniently measured by electron paramagnetic resonance (EPR) spectroscopy. Powdered enamel tissue exhibited EPR signals remarkably similar to the radicals formed by ionizing radiation. The observations described below lead to the conclusion that physical stress will induce a free-radical formation in dental tissues. These observations have significance for other areas of study such as dosimetry and archeological dating. PMID- 2559054 TI - ESR dating of elephant teeth and radiation dose rate estimation in soil. AB - Chemical analysis of 238U, 232Th and 40K in the dentine as well as enamel of elephant tooth fossil has been carried out in order to estimate the internal absorbed dose rate of the specimens, which was estimated to be (39 +/- 4) mrad/y on the assumption of early uptake model of radionuclides. The external radiation dose rate in the soil including the contribution from cosmic rays was also estimated to be (175 +/- 18) mrad/y with the help of gamma-ray spectroscopic techniques of the soil samples in which the specimens were buried. The 60Co gamma ray equivalent accumulated dose of (2 +/- 0.2) x 10(4) rad for the tooth enamel gave "ESR age" of (9 +/- 2) x 10(4) y, which falls in the geologically estimated range between 3 x 10(4) and 30 x 10(4) y before the present. PMID- 2559055 TI - Identification of irradiated foods--methods, development and concepts. PMID- 2559056 TI - The ESR detection of irradiated food. AB - Previous work has shown that the calcified tissues in several foods give rise to characteristic ESR spectra on irradiation. Further foods have now been examined. Mussel and crab shells give large signals, compared with bones of poultry, beef or frog, while prawn cuticle gives a smaller signal. The limits of detection of irradiation vary between species but are below the doses likely to be used commercially. Quantitative estimation of dose can be achieved by re-irradiation and extrapolation to zero signal. PMID- 2559057 TI - ESR analysis of irradiated frogs' legs and fishes. AB - Electron spin resonance (ESR) spectral analysis of different parts (bones, scales, jaw, etc.) from ionized (irradiated) frozen frogs' legs and fishes (brown trout and sardine) were recorded. There is always present, after treatment, a signal due to the irradiation. ESR and ENDOR experiments lead us to assign it to h1 centers from hydroxyapatite, as in the case of other irradiated meat bones. The use of ESR to prove whether one of these foods has been irradiated or not is discussed. PMID- 2559059 TI - EPR detection of free radicals in (I) coffee and (II) gamma-ray irradiated foodstuffs. PMID- 2559058 TI - Quality assessment of coffee beans with ESR and gamma-ray irradiation. AB - Peroxy radical formation in raw coffee beans of different qualities and origins from all over the world has been studied with electron spin resonance (ESR) analysis. The gamma-ray equivalent absorbed dose (ED) which creates the same concentration of radicals is obtained by the additive gamma-ray irradiation of the coffee beans. The ED and the cup quality is somewhat inversely related suggesting that the peroxidation of the unsaturated fatty acid is somewhat indicative of the degree of the aromatic decomposition and rancidity. PMID- 2559060 TI - ESR dosimetry and applications. Proceedings of the 2nd international symposium, 10-13 October 1988, Munich/Neuherberg. PMID- 2559062 TI - Elimination of contaminant Kryptofix 2.2.2 in the routine production of 2 [18F]fluoro-2-deoxy-D-glucose. AB - Radiopharmaceutical solutions of 2-[18F]fluoro-2-deoxy-D-glucose (2-[18F]FDG) prepared via an aminopolyether-supported nucleophilic-substitution mechanism were analyzed for contaminant 4,7,13,16,21,24-hexaoxa-1,10-diazabicyclo-[8,8,8] hexacosane (Kryptofix 2.2.2). Washing the C18-immobilized [18F]fluoro tetraacetylated intermediate with 10 mL 0.1 M HCl was found to remove impurity Kryptofix 2.2.2 from the final product. Inclusion of this synthetic step allowed the robotic production of drug-quality 2-[18F]FDG in 52-56% radiochemical yield within 75 min. A thin-layer chromatographic system for the clinical screening of the radiochemical and chemical purity of this radiopharmaceutical is described. PMID- 2559061 TI - Heavy-ion-induced free radical formation in solid DNA-constituents: quantitative and structural aspects. AB - Electron spin resonance (ESR) spectroscopy was used to study free radical formation in solid, polycrystalline pellets of DNA-constituents. Dose-yield curves were obtained at low (ca 90 K) and ambient temperatures, and were analyzed for initial G-values (radicals per 100 eV absorbed energy). At both temperatures, values of one or more orders of magnitude below the corresponding x-ray values were found. Combination of x-ray and heavy-ion irradiation indicated an enhanced radical destruction as probable cause of reduced G-values. Structural aspects as analyzed from the ESR-powder spectra revealed differences in initial, low temperature radical population and in decay reactions upon annealing between heavy-ion bombardment and low-LET irradiation. PMID- 2559064 TI - Characterization and in vivo distribution of 99mTc- and 111In-labelled magnetite. AB - Magnetite (Fe3O4) particles 0.1-0.5 microns in diameter doped with either 99mTc or 111In have been prepared and characterized by electron microscopy and Mossbauer spectroscopy. The time courses of their distributions after injection into rats were determined, including studies by gamma-camera imaging; the main targets were the liver and lung. The radiolabelled magnetic agents may be of value in research studies combining radio- and magnetic resonance imaging procedures. PMID- 2559063 TI - Design, synthesis and 64Cu labeling of fatty acid analogs containing dithiosemicarbazone chelate. AB - For the development of 62Cu labeled fatty acid analogs, two fatty acid analogs, containing dithiosemicarbazone (DTS) molecule as the 62Cu coordinating site, were designed and synthesized: a fatty acid analog containing DTS molecule at the omega-position, (a) the 12,13-dioxotetradecanoic acid di(N-methyl thiosemicarbazone) (FA-DTS), and an omega-phenyl fatty acid analog containing DTS molecule at the para-position, (b) the p-carboxyundecylphenylglyoxal-di (N methylthiosemicarbazone] (PFA-DTS). FA-DTS was synthesized by the reaction of ethyl diethoxyacetate with ethyl 11-bromonundecanate by successive decarboxylation and hydrolysis and final condensation with N methylthiosemicarbazide. PFA-DTS was synthesized by the Friedel-Craft acylation of ethyl 11-phenylundecanate, selenium oxidation of the acetophenone derivative, followed by the condensation with N-methylthiosemicarbazide. Radiolabeling of FA DTS and PFA-DTS with [64Cu]copper acetate was simple, rapid and quantitative. When injected into mice, both compounds were distributed and retained in the myocardium. These results offer a good basis for further development of 62Cu labeled fatty acid analogs. PMID- 2559065 TI - Direct electrophilic iodination of 2-thiouracil using Iodo-Gen. AB - 5-Iodo-2-thiouracil (ITU) is of interest due to its ability to bind specifically to the pigment melanin during melanogenesis and is of potential value in the diagnosis and treatment of malignant melanoma. Radioiodinated ITU was prepared directly from 2-thiouracil in a two-phase reaction using Iodo-Gen in 0.05 M phosphate buffer pH 7.0. The identity radiochemical purity and stability of the product were checked by reversed-phase high pressure liquid chromatography (HPLC). ITU labeled with 123I, 125I or 131I has been produced in millicurie amounts and isolated on a semi-preparative reversed-phase HPLC column. Production time was 2-3 h, overall radiochemical yields averaged 80%; the radiochemical purity was greater than 98%. Specific activities on the order of 20 Ci/mmol have been obtained. PMID- 2559066 TI - Labeling antibodies with copper radionuclides using N-4-nitrobenzyl-5-(4 carboxyphenyl)-10,15,20-tris(4-sulfophenyl) porphine. AB - Antibody conjugates labeled with copper-64 and -67 (64Cu and 67Cu) were prepared using the porphyrin chelator N-4-nitrobenzyl-5-(4-carboxyphenyl)-10,15,20-tris(4 sulfophenyl) porphine (N-bzHCS3P). N-bzHCS3P was chosen because it has only one carboxylate group available for activation and coupling to antibody. The conjugates were characterized with respect to (1) the location of the porphyrin on the antibody, (2) the retention of immunoreactivity, and (3) the serum stability of the amide bond linking porphyrin to antibody. These studies showed that porphyrin attachment on the antibody surface is random. The conjugates exhibited high retention of immunoreactivity and reasonable serum stability for potential application in nuclear medicine. PMID- 2559067 TI - Immunocytochemical localization of cathepsins B and H in human pancreatic endocrine cells and insulinoma cells. AB - Cathepsins B and H are representative cysteine proteinases localized to lysosomes of a variety of mammalian cells. Previous studies indicated the presence of these enzymes also in secretory granules of endocrine cells. Therefore, the human endocrine pancreas and human insulinomas were investigated by light microscopical immunohistochemistry on serial semithin plastic sections immunostained sequentially for cathepsins B or H and pancreatic hormones. Out of the four established endocrine cell types, insulin (B-) and glucagon (A-) cells showed immunoreactivities for these cathepsins. Cathepsin B immunoreactivities showed a dot-like appearance in A- and B-cells and in insulinoma cells. Immunoreactivities for cathepsin H additionally were found in cell parts containing secretory granules of B-cells and insulinoma cells. By single and double immunoelectron microscopy the dot-like immunoreactivities for cathepsin B were identified as immunoreactive lysosomes of A- and B-cells and insulinoma cells. In addition, some of the secretory granules of A- and B-cells showed cathepsin B immunoreactivities. Cathepsin H immunoreactivities showed an other pattern: they were found regularly in the secretory granules of A- and B-cells and insulinoma cells, and in lysosomes of A-cells. These findings suggest that cathepsins B and H in lysosomes of A- and/or B-cells are involved in the degradation of lysosomal constituents. In secretory granules of these cells, these cysteine proteinases may participate in the processing of the corresponding hormones from their precursor proteins. PMID- 2559068 TI - cGMP immunocytochemistry in aorta, kidney, retina and brain tissues of the rat after perfusion with nitroprusside. AB - The distribution of cyclic guanosine 3',5' monophosphate (cGMP) producing cells in various organs of the rat were studied immunocytochemically using antibodies raised against formaldehyde-fixed cGMP. Sodium nitroprusside (SNP), a direct activator of guanylate cyclase and vasodilator, was used to enhance cGMP levels. In order to reach all organs optimally, whole body perfusion was performed using a modified Krebs-Ringer buffer at 37 degrees C, aerated with 5% CO2/95% O2, also containing isobutyl methyl xanthine (IBMX); a phosphodiesterase inhibitor. After 15-min pre-perfusion, SNP was added to the perfusate, followed by fast fixation with ice-cold 4% paraformaldehyde-phosphate buffer. After vehicle perfusion, only the retina showed cGMP immunoreactivity in the photoreceptor and ganglion layer, while other organs lacked cGMP immunoreactivity. After 15-min perfusion with SNP (10 microM), enhanced cGMP immunostaining was seen in smooth muscles of the aorta, amacrine-like cells in the retina, glomeruli of the kidney cortex, blood vessels in the dura mater, as well as cells in the pineal and in the median eminence. The results indicate that the distribution and the reactivity of cGMP producing cells, situated outside the blood brain barrier, can be studied by immunocytochemistry after pharmacological manipulations of the intact tissue with a nitrovasodilator using whole body perfusion. PMID- 2559071 TI - Analysis of plasmid profiling as a method for rapid differentiation of food associated Clostridium perfringens strains. AB - Plasmid analysis of over 120 strains of Clostridium perfringens, isolated during food-poisoning incidents and from animal carcasses and food constituents with no association with food poisoning, showed the potential of plasmid profiling as a means of differentiating epidemiologically related strains. On average 65% of freshly isolated strains contained one or more plasmids which could be used in the analysis. Comparison of profiles of strains from unrelated sources or unrelated strains from the same source showed a particularly wide variety of plasmid profiles. Thus the possibility that epidemiologically-unrelated strains might possess similar profiles appears to be very low in this organism. Analysis of serologically-related strains from the same source revealed similar plasmid profiles in all the plasmid-bearing strains examined. A high proportion (71%) of fresh and well-characterized food-poisoning strains possessed plasmids of 6.2 kb in size (compared with 19% of non-food-poisoning strains). The possible role of these plasmids is discussed, since the structural gene encoding the enterotoxin type A was not present on any of the plasmids in the food-poisoning strains tested. PMID- 2559070 TI - Diacylglycerols enhance the anti-tumor effect of glucocorticoid on L5178Y lymphoblasts in vivo. AB - 1,2-Dioleoyl-rac-glycerol, a potent activator of protein kinase C, was found to enhance the growth-inhibitory effect of triamcinolone acetonide on L5178Y lymphoblasts in adrenalectomized, male DBA/2 mice. On the other hand, in mice without adrenalectomy, it markedly inhibited tumor growth without increasing the plasma level of corticosterone or adrenocorticotropic hormone or reducing the body weight. These results suggest that diacylglycerol enhances the action of endogenous glucocorticoid to a sufficient level to inhibit the growth of lymphoblasts. Of various diacylglycerols with different carbon chain lengths tested, 1,2-dioleoyl-rac-glycerol was the most potent growth inhibitor and was maximally effective at a dose of above 30 micrograms/100 g body weight. This finding suggests that diacylglycerols may be useful for enhancing the antitumor effect of a low dose of glucocorticoid or endogenous glucocorticoid on lymphoblasts without any significant side effect. PMID- 2559069 TI - Comparative study of placental protein 19, human chorionic gonadotrophin and pregnancy-specific beta 1-glycoprotein as immunohistochemical markers for extravillous trophoblast in pregnancy and trophoblastic disease. AB - PP19, a new placental tissue protein, has alpha 1-beta 1 electrophoretic mobility, a molecular weight of 36,500 and 3.9% carbohydrate. To study immunocytochemical PP19 localization in extravillous trophoblast, we obtained formalin-fixed specimens from extravillous tubal pregnancy at gestational weeks (GW) 7-9 (12 blocks); four early intrauterine pregnancies at GW 7-13 (12 blocks); four late pregnancies at GW 28-38 complicated with intramural uterine myoma, placenta increta and abruptio placenta (8 blocks); four invasive complete moles (9 blocks); and seven primary and metastatic gestational choriocarcinomas (12 blocks). Immunohistochemical staining was done for PP19, pregnancy-specific beta 1-glycoprotein (SP1) and human chorionic gonadotrophin (hCG) using the indirect labeled antibody method [purified PP19 (Lot no. 225/242) and antibody against PP19 (Lot no. 632ZA) prepared by H. Bohn, antibodies against hCG (Behringwerke, Marburg, FRG) and SP1 (Dakopatts, Copenhagen, Denmark)]. In both early and late intrauterine pregnancies, the extravillous syncytiotrophoblastic cell (XST) showed positive staining for hCG and SP1 in the cytoplasm, as well as for PP19, which stained more intensively in the nucleus than in the cytoplasm. The three proteins were not seen in the evtravillous cytotrophoblastic cell (XCT) in the trophoblastic cell column and shell. The interstitial cytotrophoblast-like cell (ICT), which infiltrated into the decidua and myometrium, and their blood vessels, was immunoreactively positive for PP19 but negative for hCG and SP1 with the exception of SP1-positive ICT in the myometrium in late pregnancy. XST and ICT in the endosalpinx of tubal pregnancy stained for all three proteins.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559072 TI - Comparison of the in-vitro effect of several macrolides on the oxidative burst of human neutrophils. AB - We have compared the in-vitro interaction of five macrolides (roxithromycin, erythromycin, spiramycin, oleandomycin and josamycin) with human neutrophils (PMN). Only roxithromycin strongly impaired the oxidative burst of PMN assessed by luminol amplified chemiluminescence, superoxide anion generation, and myeloperoxidase-mediated iodination of proteins. This effect was observed only for high concentrations of this drug (100 and 50 mg/l). Furthermore, the sensitivity of PMN to the depressive effect of roxithromycin permitted the definition of two kinds of PMN: in Highly Sensitive (HS)-PMN, the oxidative response was completely abolished while in Moderately Sensitive (MS)-PMN, a decreased, but yet measurable (20-50% of the control), response was obtained. The roxithromycin-induced depression of PMN was time-dependent and partly reversed by washing. Chemotaxis was also impaired by roxithromycin (100 mg/l) but phagocytosis of Klebsiella pneumoniae was unaltered even at high concentrations of the drug. Since roxithromycin displays the highest intracellular uptake, compared with the other macrolides assessed in this study, this could explain the results observed here. The relevance to the clinical situation needs further study. This effect of roxithromycin could be useful to control the inflammatory process associated in certain infectious diseases, in particular if high concentrations of the drug are obtained in tissues. PMID- 2559073 TI - High fibre diet-methi seeds--in diabetes mellitus. PMID- 2559074 TI - Ultrasonic detection of Budd Chiari syndrome in hepatoma. AB - A case of Budd Chiari syndrome in a 50 years female due to hepatoma is reported, which was worked up ultrasonically. A brief review of literature is also presented. PMID- 2559075 TI - Long term follow-up of depot iodine in goitre. PMID- 2559076 TI - Rapid purification and structural study of DNA topoisomerase I from human Burkitt lymphoma Raji cells. AB - We have developed a rapid purification method for DNA topoisomerase I from Raji cells, a human Burkitt lymphoma cell line, using ammonium sulfate fractionation followed by chromatography on a Mono S column (FPLC, Pharmacia). By this method, the enzyme could be purified to near homogeneity within one day. Electrophoresis on sodium dodecyl sulfate polyacrylamide gel revealed that the final preparation is mainly composed of a 100-kDa protein. The major enzyme activity sedimented through a glycerol density gradient at 5.7S, accompanied with a minor peak at 8.7S. The former may correspond to the monomer of the 100-kDa polypeptide, and the latter, to its dimeric form. The gel filtration study of the crude extract revealed an active molecular species of 200 kDa, in addition to 100 kDa, and lower molecular weight forms. These results suggest that DNA topoisomerase I is largely in monomeric form, but also has a minor population of the dimeric form. PMID- 2559077 TI - Ca-release by phosphoinositides from sarcoplasmic reticulum of frog skeletal muscle. AB - To clarify the biological role of phosphoinositides including inositol trisphosphate (IP3) in the skeletal muscle, we examined the Ca-releasing action on the heavy fraction of sarcoplasmic reticulum (HFSR) from bullfrog skeletal muscle of IP3, phosphatidylinositol monophosphate (PIP), phosphatidylinositol 4,5 bisphosphate (PIP2), and glycerophosphoinositol 4,5-bisphosphate (GPIP2). Only PIP2 caused dose-dependent Ca release. IP3 (up to 55 microM), PIP (up to 37 microM), and GPIP2 (up to 33 microM) were ineffective. The PIP2-induced Ca release is due to the direct action of PIP2, but not its metabolite(s). The properties of the PIP2-induced Ca release are unique and cannot be accounted for by the Ca release mechanisms already reported, such as Ca2+-induced, ionic substitution-induced, or IP3-induced Ca release. The rate of the PIP2-induced Ca release, however, is so slow that it may have no physiological relevance unless stimulating factors or agents exist. PMID- 2559079 TI - Bi-directional regulation of dephosphorylation of cAMP-dependent phosphorylated proteins by cAMP and calcium in permeabilized rat heart cells. AB - We studied the regulation of dephosphorylation of cAMP-dependent phosphorylated proteins of isolated, permeabilized (skinned) myocardial cells from adult rat. Staurosporine, a potent inhibitor of protein kinase, inhibited cAMP-dependent phosphorylation of phospholamban and troponin-I, the key proteins in the control of contraction and relaxation of the myocardial cells. Staurosporine antagonized the stimulatory action of cAMP on the spontaneous beating of the myocytes accompanied by dephosphorylation of phospholamban but not of troponin-I at pCa 7 8. In cold ATP dilution experiments with apparent stoppage of protein phosphorylation, dephosphorylation of phospholamban was accelerated both by Ca2+ and staurosporine but that of troponin-I took place only in the presence of Ca2+ ion (pCa less than 6.5). These phenomena suggest a bi-directional regulation of dephosphorylation of the key proteins by the intracellular messengers cAMP and Ca2+. PMID- 2559078 TI - Mono-sulfated globotetraosylceramide from human kidney. AB - A novel sulfated glycosphingolipid that belongs to "globo-series" was isolated from human kidney. This lipid was purified from a pooled kidney preparation by chloroform-methanol extraction, mild alkaline treatment, DEAE-Sephadex and silicic acid column chromatographies, and preparative TLC. The structure and the properties were studied by IR spectroscopy, proton NMR spectroscopy, negative secondary ion-mass spectrometry, solvolysis, periodate oxidation, compositional and methylation analyses, monoclonal antibodies, and a sulfatide-binding protein. From the results of the above analyses, the structure of this glycolipid was proposed to be HSO3-3GalNAc beta 1-3Gal alpha 1-4Gal beta 1-4Glc beta 1 1ceramide. This sulfated lipid reacted with a monoclonal anti-SSEA-3 (stage specific embryonic antigen-3) (MC-631) (Kannagi, R., Cochran, N.A., Ishigami, F., Hakomori, S., Andrews, P.W., Knowles, B.B., & Solter, D. (1983) EMBO J. 2, 2355 2361), whose epitope is R-3GalNAc beta 1-3Gal alpha 1-4Gal beta 1-R', on TLC and solid-phase radioimmunoassay. This lipid also bound to the 125I-labeled sulfatide binding protein, thrombospondin. The yield of this sulfated glycolipid was 34 pmol/g of tissue, which was about 0.028, 0.16, and 18 mol% of galactosyl- and lactosylceramide sulfates, and globopentosylceramide sulfate (Nagai, K.-i., Roberts, D.D., Toida, T., Matsumoto, H., Kushi, Y., Handa, S., & Ishizuka, I. (1989) J. Biol. Chem. 264, in press), respectively, in human kidney. PMID- 2559080 TI - 3(20)alpha-hydroxysteroid dehydrogenase activity of monkey liver indanol dehydrogenase. AB - Homogeneous indanol dehydrogenase from monkey liver catalyzed the reversible conversion of 3 alpha- or 20 alpha-hydroxy groups of several bile acids and 5 beta-pregnanes to the corresponding 3- or 20-ketosteroids. The kcat values for the steroids determined at pH 7.4 were low, but the kcat/Km values for the 3 ketosteroids were comparable to or exceeded those for 1-indanol and xenobiotic carbonyl substrates. The enzyme transferred the 4-pro-R-hydrogen atom of NADPH to the 3 beta- or 20 beta-face of the ketosteroid substrate. Competitive inhibition of the hydroxysteroid dehydrogenase activity of the enzyme by medroxyprogesterone acetate, hexestrol, and 1,10-phenanthroline suggests that both 1-indanol and hydroxysteroid are oxidized at the same active site on the enzyme. The specific inhibitor of the enzyme, 1,10-phenanthroline, suppressed the 3 alpha hydroxysteroid dehydrogenase activity in the crude extract of monkey liver by 50%. The results strongly suggest that indanol dehydrogenase acts as a 3(20)alpha hydroxysteroid dehydrogenase in the metabolism of certain steroid hormones and bile acids. PMID- 2559081 TI - Action of a bacterial Achromobacter collagenase on the soft carious dentine: an in vitro study with the scanning electron microscope. AB - Samples of carious human teeth treated in vitro for 48-92 hours with a bacterial Achromobacter collagenase in solution in borate buffer at 33 degrees were examined with the scanning electron microscope. The enzyme was seen to destroy soft carious dentine but not sound layers of dentine beneath the lesion. This finding may have clinical implications. PMID- 2559082 TI - The effect of cyclodextrin on lipopolysaccharide production in cultures of Bordetella pertussis. AB - The effect of adding 500 micrograms of (2,6-0-dimethyl) beta-cyclodextrin (Me beta-CD) per ml of Stainer-Scholte (SS) medium in two-day shaker flask cultures of Bordetella pertussis on the production of lipopolysaccharide (LPS) was investigated. The amount of LPS per 10(9) cells found in the supernatants of these cultures was either somewhat reduced or unaffected by comparison with the amounts in cultures grown in SS-medium alone. In addition, the time course of LPS release from cultures of B. pertussis strain 3843 cells during a 96-h growth period in normal and Me-beta-CD-enriched SS medium is described. By using the enriched medium bacterial growth, the production of filamentous haemagglutinin (FHA) and of pertussis toxin (Pt) and the levels of haemagglutination and lymphocytosis-promoting activity were enhanced to various degrees. Measurements made on sedimented whole and on sonicated B. pertussis cells grown in the two media showed no differences in LPS content. The reasons for the reduced/unaffected LPS production are discussed. It has been suggested that an interaction between hydrophobic cavities of the Me-beta-CD molecules and the 'lipid A' part of LPS reduces the reactivity of LPS in the Limulus Amoebocyte Lysate (LAL) assay. This possibility, however, was rejected as the reactivity of Me-beta-CD-spiked purified B. pertussis strain 3803 LPS, compared with unspiked samples, remained unchanged. PMID- 2559083 TI - An international collaborative study of an anti-infectious bursal disease virus reference serum. AB - Fourteen laboratories participated in a collaborative study of a freeze-dried preparation of anti-infectious bursal disease virus serum to assess the suitability of the serum as a standard for use in the infectious bursal disease virus neutralization test. Ten laboratories carried out micro-virus neutralization tests and six carried out plaque reduction tests, two laboratories carrying out both tests. When titres were expressed as a proportion of that obtained for a reference preparation there was a marked reduction in variation between results from different laboratories. The use of a reference preparation was therefore of value when comparing results from different laboratories. It is proposed that the reference preparation used in this study be used as a standard to facilitate the comparison of results from different laboratories. The proposed standard contains by definition 10,000 UK units. PMID- 2559084 TI - Heat inactivation of human immunodeficiency virus type 2 (HIV-2). PMID- 2559085 TI - Spectrometric and chromatographic methods for the analysis of polymeric explant materials. AB - Several analytical pyrolysis methods, namely pyrolysis mass spectrometry (Py-MS), time-resolved pyrolysis mass spectrometry (TRPy-MS), and pyrolysis short column gas chromatography mass spectrometry (Py-GC/MS) were used to analyze polymers of clinical interest both before and after implantation. A sample of Biomer, a poly(ether urethane urea) used in the Utah artificial heart, was analyzed using these methods. Two poly(ether urethanes) (Tecoflex and Pellethane) and a poly(dimethylsilicone) (Silastic) sample were analyzed using Py-GC/MS. The direct Py-MS of Biomer identified the components used in the manufacture of Biomer. Py GC/MS of Biomer, Tecoflex, Pellethane, and Silastic also identified the components used in their manufacture. The analysis of explanted Biomer detected the presence of adsorbed cholestadiene, the reaction of chloride ions with a stabilizer, and the presence of a siloxane contaminant. The cholestadiene was detected on the outside housing of an artificial heart which had been implanted for 297 days. The cholestadiene was detected at low levels and was identified by library search on the MS data system. The siloxane contaminant was also identified by the MS data system. All of the methods demonstrated required only short instrumental analysis times (10 min or less). Data analysis required much more time, but much of the data analysis can be automated. PMID- 2559086 TI - Culture of primary lung tumors using medium conditioned by a lung carcinoma cell line. AB - Medium conditioned by the established lung tumor cell line A549 was used as a supplement to culture cells from primary solid lung tumors. Of 36 cases placed into culture, primary cells were obtained in 33 (91.7%). Of 29 cases in which subcultures were attempted, 18 (62.1%) were successful. Nine cell lines have been established by this technique to date. In growth assays, conditioned medium (CM) was found to stimulate both monolayer colony formation and growth in semi-solid medium of cells cultured from primary solid tumors. CM has been found to contain factors with the properties of both transforming growth factor alpha (TGF alpha) and insulin-like growth factor-I (IGF-I). The addition of a combination of these factors as purified peptides to basal medium at levels found in CM (0.1-0.5 ng/ml) stimulated colony formation of lung tumor cells by up to fourfold. These results indicate that secretion of growth factors may be important in tumor growth in vivo, and that use of CM may be a valuable tool for obtaining cultures from primary solid tumors. PMID- 2559087 TI - Changes in gap junction protein (connexin 32) gene expression during rat liver carcinogenesis. AB - A rat liver gap junction (GJ) cDNA probe that detects mRNA encoding the 32 Kd GJ protein (connexin 32) was employed to study GJ-protein gene expression in rat liver tumors induced by a single exposure to diethylnitrosamine (DEN) followed by exposure to 2-acetylaminofluorene (AAF)/CCl4/AAF or induced by systemic administration of N-ethyl-N-hydroxyethylnitrosamine (EHEN). All carcinomas generated by these carcinogens showed markedly reduced levels of GJ-protein mRNA. This may indicate that GJ-protein levels and gap-junctional intercellular communication (GJIC) capacity are also severely compromised. Moreover, all hyperplastic nodules also showed a reduced level of GJ-protein mRNA. Taken together with our earlier finding that the liver tumor promoter phenobarbital inhibits GJ-protein gene expression, these results suggest that deranged GJIC is a relatively early event in liver multistage carcinogenesis. A range of other cDNA probes was also used to characterize gene expression in the DEN-induced tumors. Induction of expression was seen for glutathione S-transferase (placental form) (GST-P), gamma-glutamyltranspeptidase (GGT), and c-raf but not for c-Ha-ras or c-myc. PMID- 2559088 TI - Identification of immunoglobulin heavy chain binding protein as glucose-regulated protein 78 on the basis of amino acid sequence, immunological cross-reactivity, and functional activity. AB - Immunoglobulin heavy chain binding protein (BiP) associates transiently with various proteins destined for the secretory pathway. To investigate the relationship between BiP and the 78K (K = 10(3) Mr) glucose-regulated protein (GRP78), we have determined a partial amino acid sequence of purified mouse BiP and isolated and sequenced a full-length cDNA clone encoding mouse GRP78. The 26 amino-terminal residues of the mature BiP protein are identical to a sequence of amino acids located near the start of the open reading frame encoding GRP78. A polyclonal antiserum raised against mouse GRP78 protein expressed in bacteria from the cloned GRP78 cDNA could immunoprecipitate complexes consisting of BiP and unfolded forms of immunoglobulin heavy chains. Furthermore, a monoclonal antibody raised against mouse BiP immunoprecipitated mouse GRP78 expressed in monkey CV-1 cells from an SV40-GRP78 recombinant vector. Finally, like the endogenous BiP of simian cells, mouse GRP78 associated with malfolded, non glycosylated forms of influenza hemagglutinin (HA) when GRP78 and HA were co expressed from SV40 vectors in CV-1 cells. These studies confirm that BiP is identical to GRP78. Comparison of the nucleic acid and deduced amino acid sequence of mouse GRP78 with those of other rodent and human GRP78s revealed an extremely high degree of sequence identity. BiP/GRP78 is closely related (approximately 60% identity) to the cytoplasmic 70K heat-shock proteins. Surprisingly, the carboxy-terminal 29 amino acids of BiP/GRP78, which are not conserved in HSP70 proteins, are almost identical in sequence to the steroidogenesis activator peptide found in the cytoplasm of rat Leydig tumor cells. Possible relationships between these polypeptides are discussed. PMID- 2559089 TI - The structure and function of a mannose 6-phosphate receptor-enriched, pre lysosomal compartment in animal cells. AB - The characteristics of a distinct prelysosomal compartment, evident in many animal cells, are described. The main feature of this compartment is that it is an acidic, tubulo-vesicular late endosome structure that contains high concentrations of the cation-independent mannose 6-phosphate receptor. The pathway to the lysosomes from the early endosomes, as well as that of newly synthesized lysosomal proteins from the trans-Golgi network, appear to converge in this prelysosomal compartment. PMID- 2559090 TI - Import of proteins into the various submitochondrial compartments. AB - Import of proteins into mitochondria can be subdivided into several distinct steps. (1) Mitochondrial proteins are synthesized on free ribosomes and are released into cytosolic pools. Nucleoside triphosphates are required to keep precursors in a conformation competent for import. (2) Precursors are directed to mitochondria by specific targeting signals (in most cases contained in N-terminal presequences) and by binding to receptors on the surface of the outer membrane. (3) Precursors interact with a component in the outer membrane which is believed to facilitate membrane insertion ('general insertion protein'). (4) Outer membrane proteins are then directly routed to their final location. Proteins of all other submitochondrial compartments are directed into translocation contact sites between outer and inner membranes. Transfer into contact sites is dependent on the membrane potential (delta psi) across the inner membrane. (5) Presequences of precursors are cleaved in the matrix by the mitochondrial processing peptidase in cooperation with the processing enhancing protein. (6) Precursors of the intermembrane space or the outer surface of the inner membrane have to be re translocated back across the inner membrane ('conservative sorting'). PMID- 2559092 TI - Nucleoside diphosphokinase, an enzyme with step changes in activity during the cell cycle of the fission yeast Schizosaccharomyces pombe. II. Dissociation of the steps from the DNA-division cycle after induction synchronization. AB - Synchrony was induced in cultures of the mitotic mutant cdc2.33 of Schizosaccharomyces pombe by shifting up an asynchronous culture to the restrictive temperature for a period of 3.5-4.5 h and then shifting down to the permissive temperature. The resulting synchronous divisions had short cycle times, down to 50% of the normal cycle. The oscillatory control of nucleoside diphosphokinase activity was also synchronized by the shift-down and the activity rose in a step pattern. Unlike the situation in the normal cycle, this step pattern was dissociated from the shortened cell cycle and had a longer period and different phase relations. It may be that the normal entrainment or coupling between the cell cycle and the activity control fails if the cell cycle is too short. The period of the activity control (equal to the protein doubling time at the restrictive temperature) appears to be temperature-compensated. PMID- 2559091 TI - Injection of Xenopus eggs before activation, achieved by control of extracellular factors, improves plasmid DNA replication after activation. AB - Injection of molecular probes into unfertilized Xenopus eggs requires suppression of activation. But the unfertilized egg is poised for activity, and pricking, like sperm penetration, triggers the start of the first cell cycle. Methods of suppressing activation generally rely on introduction of drugs into the cell, but some of these techniques are irreversible. I report here that injection without activation can also be accomplished by simply limiting extracellular free Ca2+ to 1-2 microM. The site of injection heals, but the cortex does not contract. Gentle modification of the vitelline envelope, which causes it to become tougher, improves the rate of healing to about 100%. Healed eggs are stable for hours and can be activated when needed. Injection of a plasmid derived from type 1 bovine papilloma virus revealed that replication occurs only after activation, but preloading the DNA markedly increased the efficiency of first-round replication. DNA interaction with the unactivated egg cytoplasm may therefore be required for efficient replication of exogenous DNA. The new procedures described here are likely to be of general utility. PMID- 2559093 TI - Lithium ions induce prestalk-associated gene expression and inhibit prespore gene expression in Dictyostelium discoideum. AB - We investigated the effect of Li+ on two types of cyclic AMP-regulated gene expression and on basal and cyclic AMP-stimulated inositol 1,4,5-triphosphate (Ins(1,4,5)P3) levels. Li+ effectively inhibits cyclic AMP-induced prespore gene expression, half-maximal inhibition occurring at about 2 mM-LiCl. In contrast, Li+ (1-3 mM) promotes the cyclic AMP-induced increase of cysteine proteinase-2 mRNA levels, and induces the expression of this prestalk-associated gene in the absence of cyclic AMP stimuli. At concentrations exceeding 4-5 mM, LiCl inhibits cysteine proteinase-2 gene expression. LiCl reduces basal Ins(1,4,5)P3 levels and decreases the cyclic AMP-induced accumulation of Ins(1,4,5)P3; both effects occur half-maximally at 2-3 mM-LiCl. These results indicate that the induction of the cysteine proteinase-2 gene by Li+ is not due to elevated levels of Ins(1,4,5)P3. It is, however, possible that inhibition of prespore gene expression by Li+ is caused by Li+-induced reduction of basal and/or stimulated Ins(1,4,5)P3 levels. PMID- 2559095 TI - Induction of DNA synthesis by cholera toxin in the temperature-sensitive cell cycle mutants of rat 3Y1 fibroblasts at a restrictive temperature. AB - Four temperature-sensitive mutants of rat 3Y1 fibroblasts representing separate complementation groups (3Y1tsD123, 3Y1tsF121, 3Y1tsG125 and 3Y1tsH203) are arrested at a restrictive temperature of 39.8 degrees C mainly with a G1-phase DNA content (temperature arrest). Cholera toxin (CT) (3 micrograms ml-1) induced DNA synthesis at 39.8 degrees C in the temperature-arrested cultures of two mutants (3Y1tsD123 and 3Y1tsG125). This effect of CT was not mimicked by other agents known to elevate the cellular level of cyclic AMP, such as dibutyryl cyclic AMP, prostaglandin E1 and forskolin, suggesting that the elevation of cellular cyclic AMP level per se is not responsible for the induction of DNA synthesis by CT. Addition of the B subunit of CT to the temperature-arrested cultures of 3Y1tsD123 and 3Y1tsG125 did not induce DNA synthesis at 39.8 degrees C, indicating that the binding of CT to the cell surface alone is insufficient for the induction. The CT-treated cell membrane fraction prepared from temperature-arrested 3Y1tsG125 cells had similar activity for [32P]ADP ribosylation of the 45 X 10(3) Mr protein to that prepared from cells proliferating at a permissive temperature of 33.8 degrees C. All these results suggest that 3Y1tsG125 cells utilize a CT-responsive signal transduction pathway, different from adenylate cyclase cascade, for preparation for entry into S phase in the temperature-arrested 3Y1tsG125.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559094 TI - Immunochemical localization of amiloride-sensitive sodium channels in sodium transporting epithelia. AB - The localization of amiloride-sensitive Na+ channels in Na+-transporting epithelia was examined using antibodies made against amiloride-binding Na+ channel protein purified from bovine kidney. The distribution of the channel protein was determined in thick frozen sections at the light-microscopic level using indirect immunofluorescence, and at the electron-microscopic level using immunogold labelling. In the cells of both the intact bovine collecting tubule and A6 confluent monolayers, only the luminal or apical-facing surface membranes showed staining. Sodium channel protein was characteristically localized on microvillar domains of the apical plasma membrane. Little or no basolateral membrane staining was evident. Channel protein was also absent from subapical vesicles and tight junctions, and was not found in bovine renal proximal tubules, cultured human secretory sweat coils, non-epithelial Chinese hamster ovary (CHO) cells or human skin fibroblasts. Trypsinization of intact A6 monolayers prior to cell fixation abolished specific staining with antibody. Pretreatment with amiloride protected against this loss of staining. Thus, our probes are specific for amiloride-binding Na+ channel protein, and this channel protein is largely or completely confined to the apical membrane of Na+-transporting epithelia. The level and distribution of specific immunostaining in A6 cells was unchanged by aldosterone treatment, although channel activity, as measured by short-circuit current, increased threefold. This result demonstrates that Na+ channel protein is ever present at the cell surface and exists in both an active and an inactive form. We find no evidence that stimulation of Na+ uptake by aldosterone involves recruitment of new channels from a cytoplasmic pool. PMID- 2559096 TI - Chromatographic method for the preparation of apo-cellular retinol-binding protein and apo-cellular retinoic acid-binding protein from their holo-types. AB - A chromatographic method is described for the preparation of apo-cellular retinol binding protein (CRBP) and apo-cellular retinoic acid-binding protein (CRABP) from their corresponding holoproteins. Elimination of retinoids from either purified CRBP or CRABP holoprotein complex could be performed quantitatively by DEAE-cellulose chromatography without any alteration in the inherent properties of the native proteins. In contrast, the usual methods, involving UV irradiation or acetone precipitation, resulted in some modification of these binding proteins. This chromatographic method was also applicable to the preparation of apo-fatty acid-binding protein (FABP) from FABP-palmitic acid holoprotein complex. PMID- 2559098 TI - Influence of perchloric acid on ion-pair high-performance liquid chromatography of nucleotides. PMID- 2559097 TI - Method for the determination of the specific activities of UTP and CTP in mouse kidney by high-performance liquid chromatography. PMID- 2559099 TI - Non-isotopic detection of HPV DNA in cervical smears using dot-blot hybridization. AB - Infection of the cervix with specific types of the human papillomavirus (HPV) results in condylomatous, dysplastic and/or neoplastic epithelial changes. To enable routine screening of premalignant cervical lesions for HPV DNA, we have developed a sensitive method of detecting HPV -6, -11, -16, -18 and -33 in cervical scrapes using a non-isotopic, dot-blot hybridization assay. Cloned, genomic HPV DNA is labelled with biotin-11-dUTP by nick translation, and used to screen for HPV types under high stringency conditions. DNA is extracted from cervical cells and spotted on nitrocellulose membranes. Cervical cells, from 20 patients attending colposcopy were screened for HPV types. All patients had a dysplasia or condyloma. HPV DNA was isolated from 80%. The method visualized an amount of target DNA as low as 1 pg without background. Compared with Southern blot hybridization utilizing radiolabelled probe, this assay is equally sensitive and specific. The technique is easy to do, and may significantly contribute to the management of cervical dysplasia. PMID- 2559101 TI - Increased sensitivity for rapid detection of cytomegalovirus by shell vial centrifugation assay using mink lung cell cultures. AB - A comparative study was made of various human and non-human cell cultures to determine their sensitivity for cytomegalovirus (CMV) as detected by the production of CMV early antigen using the shell vial centrifugation assay. Mink lung cell cultures, frequently used for detection of herpes simplex virus in clinical specimens, were found to be significantly more sensitive to infection by CMV than other cell cultures tested. Using the shell vial centrifugation assay, the mink lung cell cultures were more sensitive than human diploid fibroblasts for the detection of the Davis strain of human CMV and CMV from clinical specimens. PMID- 2559100 TI - Enzyme-linked immunosorbent assay for IgG and IgM antibodies against human parvovirus B19: use of monoclonal antibodies and viral antigen propagated in vitro. AB - Enzyme-linked immunosorbent assay (ELISA) systems for serum IgG and IgM antibodies to human parvovirus B19 were established by utilizing anti-B19 monoclonal antibodies (mAbs) and human plasma B19 antigen. The specificities of IgG and IgM ELISA were confirmed by indirect immunofluorescence staining and Western blot immunoassay with panel sera. The series of serum specimens obtained from two B19-infected patients were examined with ELISA. The IgM antibody titers increased quickly after the onset of the symptoms and returned to a negative range after five months. The IgG antibody titers also increased just after the increasing of IgM titers and the elevated levels continued for more than a year. We also established the same ELISA systems by utilizing in vitro propagated B19 antigen and similar results were obtained. PMID- 2559102 TI - Parainfluenza virus infection of cultured airway epithelial cells. AB - Studies of the cellular effects of respiratory viruses have generally used cultures of non-airway (particularly renal) epithelial cells. This requires the assumption that, despite the marked differences between renal epithelium and airway epithelium, the virus-host cell interactions in cultures of renal epithelium will be relevant to those in airway epithelium. To study viral infection of airway epithelial cells, we removed the epithelial cells from ferret tracheas using 0.1% pronase solution, and plated them at a density of 5 X 10(5) cells/cm2 in collagen-coated plastic tissue culture wells. Cultures grew to confluence after 5-7 days. Viral inocula, consisting of supernatants from parainfluenza type 1-infected rhesus monkey kidney cell monolayers, were added to the culture medium in a concentration 10(3) times that sufficient to produce infection in 50% of rhesus monkey kidney monolayers (TCID50). Cytopathic changes, consisting of cellular elongation and detachment, became apparent after 3-6 days, at which time the medium contained 5 X 10(8) TCID50/ml. The monolayer appeared to be uniformly infected as revealed by adsorption of guinea pig erythrocytes. Specific immunofluorescence revealed uniformly positive staining for parainfluenza type 1 antigens. The ability to infect pure cultures of airway epithelial cells with viruses will allow us to examine the effects of these viruses on epithelial cell function, and to study virus-host cell interactions in cell cultures derived from the natural host cell. PMID- 2559103 TI - A method for the enumeration of poliovirus in selected molluscan shellfish. AB - A virus extraction procedure was developed and evaluated on five commercially important molluscan shellfish species: Crassostrea virginica (Eastern oyster), Mya arenaria (softshell clam), Mytilus edulis (blue mussel), Mercenaria mercenaria (hardshell clam), and Crassostrea gigas, (Pacific oyster). Shellfish tissue homogenates were spiked with poliovirus, extracted, and plaque assayed. Mean virus recoveries were: C. virginica, 63.8%; M. arenaria, 42.1%; M. edulis, 67.3%; M. mercenaria, 48.3%; and C. gigas 10.1%. Shellfish were also allowed to accumulate poliovirus from spiked seawater (10 to 20 PFU/ml of water) over 48 to 72 h. The results indicate that poliovirus could be extracted from four shellfish species exposed to near environmental levels of virus. Virus recoveries per gram of tissue were: M. arenaria, 11.7 PFU; M. mercenaria, 26.0 PFU; M. edulis, 21.5 PFU; and C. virginica, 2.0 PFU. The results of this study indicate that the procedure is effective in extracting ingested viruses from several shellfish species. This procedure may have practical application for enumeration of enteric viruses in environmental samples. PMID- 2559104 TI - A simplified procedure for oligonucleotide fingerprint analysis of multiple bluetongue virus genome segments utilizing small gels. AB - The technique of oligonucleotide fingerprinting is a useful tool for analyzing sequence homology among RNA molecules. A rapid method for the simultaneous production of multiple fingerprints has been developed using a commercially available electrophoresis apparatus. The system makes use of relatively small gels, yielding fingerprints that when compared to conventional systems are of reduced size but of comparable resolution. The system described should have particular application to the analysis of RNA viruses with multiple genome segments, and in epidemiologic studies concerned with the analysis of multiple virus isolates. PMID- 2559105 TI - Rapid sampling of multiple enzyme reactions. AB - A simple method of initiating and sampling six simultaneous reactions was devised. A commercially available vial rack was fitted with a Plexiglas overlaying sheet to stabilize the vials for the addition and sampling procedures. Glass vials were routinely used because of their thermal conductivity advantages. Samples were added and removed, and the reactions were mixed with a multichannel pipet using every other channel. The data showing six simultaneous progress curves for the rapid inactivation of herpes simplex virus ribonucleotide reductase were presented and analyzed. In addition, the time course of 12 reactions catalyzed by varicella zoster virus thymidine kinase were assayed at one min intervals generating 96 data points within 8.5 min. A second experiment generated data points every 30 s for six simultaneous replicate thymidine kinase reactions. The ease of use and high reproducibility of the method are demonstrated by these data. PMID- 2559106 TI - Effect of gonadotrophin-releasing hormone and related analogue on human luteal cell function in vitro. AB - The possible direct effect of gonadotrophin-releasing hormone (GnRH) and GnRH agonist (GnRH-A; buserelin) on basal and human chorionic gonadotrophin (HCG) stimulated progesterone (P) and cyclic AMP (cAMP) production by cultured human luteal cells was examined. Luteal cells from the early or mid-luteal phase were incubated in long-term cultures. They responded to HCG stimulation with a 2- to 3 fold increase in P production and a 2-fold increase in cAMP production. The addition of GnRH (10(-7) and 10(-5) M) or GnRH-A (10(-7) and 10(-5) M) to the medium had no effect on either basal or HCG-stimulated secretion. These results indicate that both GnRH and GnRH-A have no direct effect on human luteal steroidogenesis in vitro. PMID- 2559107 TI - Isolation and characterization of gelsolin from cultured BHK cells. AB - The Ca2+-dependent actin-polymerization nucleating protein of the cytoplasmic fraction of Baby hamster kidney (BHK) C13 cells has been isolated by anion exchange, hydroxyapatite and gel-filtration chromatography. This protein has been identified as a cytoplasmic gelsolin by the following criteria: molecular mass of 90 kDa on SDS-PAGE, immunocrossreactivity with pig plasma gelsolin and similar actin-binding properties to gelsolins purified from other sources. BHK gelsolin forms a 1:2 ternary complex with rabbit muscle actin that is dependent on the presence of Ca2+. The ternary complex is dissociated on chelation of Ca2+ with EGTA to a binary complex and free actin. BHK gelsolin nucleates the polymerization of pyrene-labelled G-actin in a Ca2+-dependent manner. The proportion of unpolymerized monomer is increased in the presence of BHK gelsolin by an amount consistent with capping of the positive filament ends. The rate of actin depolymerization induced by diluting F-actin to below its critical concentration (Cc) is unaffected by the presence of BHK gelsolin in EGTA. However, in the presence of Ca2+ the rate of depolymerization is increased indicating that BHK gelsolin severs actin filaments in a Ca2+-dependent manner. PMID- 2559108 TI - Cytomegalovirus and autoimmune liver disease. PMID- 2559109 TI - Autoradiographic localization of thyrotropin-releasing hormone and substance P receptors in the rat dorsal vagal complex. AB - We utilized quantitative autoradiography to localize receptors for thyrotropin releasing hormone (TRH) and substance P in individual subnuclei of the rat nucleus tractus solitarii (NTS) and the dorsal vagal complex. Within the NTS, TRH receptor concentrations were highest within the gelatinosus and centralis subnuclei and the medial subnucleus rostral to the area postrema, moderate within the intermediate subnucleus and the medial subnucleus adjacent to the area postrema, and low within the ventrolateral and commissural subnuclei and the medial subnucleus caudal to the area postrema. In contrast, substance P receptor concentrations were high throughout the medial subnucleus, moderate in all other subnuclei medial to the tractus solitarius, and relatively low in subnuclei lateral to the tractus solitarius. The dorsal motor nucleus of the vagus contained high concentrations of both TRH and substance P receptors, whereas we observed low TRH and moderate substance P receptors in the area postrema. High TRH and moderate substance P receptors were observed in the adjacent hypoglossal nucleus. In addition, we compared the concentrations of TRH receptors between chloroform-defatted and nondefatted tissue sections, and noted little effect of white matter tritium quench upon the observed TRH receptor concentrations. These results suggest that neurotransmitter receptors within the rat dorsal vagal complex are organized in a manner consistent with previous cytoarchitectural and hodological partitioning of the NTS and that the distribution of an individual neurotransmitter receptor in the NTS may correspond to the role of that transmitter in modulating autonomic function. PMID- 2559110 TI - Immunohistochemical localization and biochemical characterization of nerve growth factor receptor in adult rat brain. AB - The expression of nerve growth factor (NGF) receptor in adult rat brain was studied by immunohistochemistry with a specific anti-rat NGF receptor monoclonal antibody, 192-IgG. Intense NGF receptor immunoreactivity (NGFRI) was found in structures known to be NGF responsive, including forebrain cholinergic neurons in medial septum, diagonal band of Broca, and basal nucleus of Meynert; central processes of neural-crest-derived sensory ganglion neurons and their innervated nucleus also contained such immunoreactivity. Distinct NGFRI staining was also found in many brain areas and cell types not known to be NGF responsive, including some hypothalamic regions, circumventricular organs, some areas related to the optic system, olfactory glomeruli, ependymal and subependymal cells in some locations, mesencephalic nucleus of the trigeminal nerve, cerebellar molecular layer, central linear nucleus, solitary tract and its nucleus, and inferior olive. The NGFRI in the circumventricular organs was further studied by in vivo labeling of 125I-ligands. Intravenously injected 125I-NGF, but not 125I cytochrome c, was specifically accumulated in the area postrema. Biochemical study of the NGF receptor showed a major band of molecular weight of approximately 90 KDa in the area postrema, choroid plexus, median eminence, and medial septum with the relative content consistent with that seen by immunohistochemistry. No evidence of a truncated NGF receptor was observed. The results of this study suggest that NGF and its receptor have broader roles in adult mammalian brain than previously thought. PMID- 2559112 TI - Type VII collagen is a component of cylindroma basement membrane zone. AB - Cylindroma basement membranes (BMs) were studied by electron microscopy and immunofluorescence with antibodies to Types IV and VII collagen in order to correlate the ultrastructure with the biochemical characteristics. Type IV collagen served as a marker for lamina densa structures and Type VII collagen for the anchoring fibrils. Ultrastructurally, the cylindroma BMs were composed of a wide band of BM-like material in which numerous anchoring fibrils were embedded. The BMs surrounding the tumor cell clusters and nodules of BM-like material entrapped within the tumor cell islands stained positively for both Types IV and VII collagen. We conclude, that anchoring fibrils are a major component of cylindroma BM zone. A trichoepithelioma investigated as control showed a BM similar to the one found at the dermoepidermal junction, i.e. a BM distinctly different from the one of cylindroma. The trichoepithelioma BMs and nodules of BM like material included within the tumor also stained positively for both Type IV and VII collagen. PMID- 2559111 TI - Papillomavirus antigen in the epidermoid cyst of the sole. Immunohistochemical and ultrastructural study. AB - An epidermoid cyst of the sole was studied immunohistochemically and ultrastructurally. Papillomavirus particles were present in the horny layer and in the upper layers of the epidermis of the cyst and within the acrosyringeal epithelium overlying the cyst. Thickened basal lamina-like structures similar to those found in the eccrine sweat duct tumors such as in cylindroma and eccrine spiroadenoma existed at the epidermal-dermal junction of the cyst wall. Carcinoembryonic antigen was immunohistochemically demonstrated in the center of the epidermoid cyst and the DAB reaction products took the shape of a circle resembling that of the sweat ducts in the horny layer. PMID- 2559113 TI - Effects of intrauterine exposure to parathion on the activity of renal ATPases in offspring. AB - The effects of ethyl parathion on the activities of various renal enzymes were studied in the offspring from dams treated with this insecticide during pregnancy. The enzymes tested were the (Na+-K+)- and the Mg2+-dependent ATPases, the glutathione S-transferases and carboxylesterases. The postnatal effects of parathion on kidney ATPases from undernourished rats were also assessed. The organophosphate was administered per os to pregnant rats at a dose of 1 mg kg-1 body weight per day throughout gestation, and suspended after delivery. The offspring were divided in groups of normally-fed and undernourished rats. In the undernourished group, food restriction produced a decrease of 43% in body weight as compared to the normally-fed group. Offspring were sacrificed 6 weeks after birth and the enzymatic activities were determined in kidney homogenates. We found a decrease in the enzymatic activity of total ATPases, at the expense of the Mg2+-dependent ATPase. However, the activities of the (Na+-K+)-dependent ATPase, the glutathione S-transferases and the carboxylesterases did not show significant changes. On the other hand, undernutrition did not potentiate the effects of parathion on the ATPases. Thus, this organophosphate administered during pregnancy produced a selective inhibition on the renal Mg2+-dependent ATPase from offspring, which was not potentiated by our undernutritional model. PMID- 2559114 TI - Neutrophil superoxide production measurement by means of stable nitroxide radical accumulation detected by ESR. AB - A new assay for superoxide radicals is based on the interaction of hydroxylamine (1-oxy-2,2,6,6-tetramethyl-4-oxopiperidine) with superoxide, giving rise to a stable nitroxide radical. Working concentration ranges of hydroxylamine and cells are determined. It was shown that the amount of superoxide generated was proportional to the concentration of nitroxide radicals. The sensitivity and specificity of the proposed assay were compared to chemiluminescence and cytochrome-c reduction. PMID- 2559115 TI - Control and ontogeny of hypothalamic-pituitary-adrenal function in the fetal rat. PMID- 2559116 TI - Continuous central vasopressin infusion increases peripheral fetal corticotropin and cortisol in the fetal sheep. AB - In previous studies on regulation of fetal adrenocorticotropin (ACTH) secretion, corticotropin releasing factor (CRF) and arginine vasopressin (AVP) have been administered by peripheral intravascular infusion. In order to look at an alternate route of administration, we investigated the effect of continuous intracerebroventricular administration of AVP to the fetus on fetal plasma ACTH and fetal and maternal plasma cortisol concentrations. Sheep fetuses (n = 9) were instrumental with carotid artery and lateral cerebral ventricular catheters. Fetuses were given intracerebroventricular infusion from 125-134 days gestational age of artificial cerebrospinal fluid vehicle (n = 4), or AVP 250 mu U.min-1 continuously in artificial cerebrospinal fluid vehicle (n =5). Fetal blood was obtained daily between 09.00 and 12.00h and 20.00 and 23.00h. Over the infusion period, fetal plasma ACTH and cortisol concentrations in AVP infused fetuses increased (P less than 0.05) compared with the vehicle infused group. Gestation length for the fetuses in the AVP and vehicle infused groups were 139 +/- 4.9 (n =4) and 145 +/- 4.6 (n = 3) days respectively (n.s.). Fetal plasma AVP concentrations in the AVP infused group were not different from the vehicle infused group. PMID- 2559117 TI - In vivo regulation of adrenocorticotrophin secretion in the immature ovine fetus. Modulation by ovine corticotrophin releasing hormone and arginine vasopressin. AB - This study investigates the in vivo regulation of ACTH secretion in the immature ovine fetus by AVP and oCRH. Previously we have demonstrated that whilst AVP containing neurones are present from 42 days, oCRH-containing neurones cannot be detected in the fetal paraventricular nucleus or median eminence until after 90 or 100 days respectively. In acutely exteriorized fetuses aged between 64-90 days (n = 5), a haemorrhagic stress elicited a significant increase (P less than 0.01) in ACTH values. There was also a significant correlation between plasma ACTH and AVP concentrations in these fetuses. In chronically cannulated fetuses less than 100 days (n = 6) injection of AVP (200 ng) significantly elevated fetal plasma ACTH values at 10 min (P less than 0.01) post injection. Simultaneous injection of AVP (200 ng) and oCRH (10 micrograms) into these fetuses produced a plasma ACTH value that was significantly greater at 10 min (P less than 0.05) than the summed response obtained with separate injection of oCRH and AVP. When AVP and oCRH were injection in equimolar amounts to fetuses between 101-118 days, AVP (2 micrograms) was found to have a greater effect on ACTH than was oCRH (2 micrograms). Pretreatment with a specific vascular antagonist of AVP d(CH2)5Tyr(Me)AVP failed to significantly inhibit the increase in mean arterial pressure associated with AVP (2 micrograms) injection but partially antagonized (P = 0.04) the decrease in fetal heart rate. The antagonist however completely abolished any effect of AVP on fetal plasma ACTH values. This study suggests that AVP most likely acts through receptors in the fetal pituitary with V1 characteristics. PMID- 2559118 TI - Entamoeba histolytica and other parasitic infections in south Kanara district, Karnataka. AB - Single stool specimens collected from 1,020 apparently healthy people of the South Kanara District, were processed for intestinal parasites using three parasitological methods viz. (a) Direct smear in physiological saline and D'Antoni's iodine, (b) Zinc sulfate concentration method and (c) by culture in modified Boek and Drbolhav medium. Of these 781 (76.51 per cent) were found to be infected with parasites. The prevalence of various intestinal parasites was: Ascaris lumbricoides (48.33 per cent), Necator americanus (46.86 per cent), Trichuris trichiura (42.75 per cent), Entamoeba coli (23.24 per cent), Entamoeba histolytica (7.94 per cent), Enterobius vermicularis (2.84 per cent), Giardia intestinalis (2.45 per cent), Iodamoeba buitschlii (1.57 per cent), Entamoeba hartmanni (1.37 per cent), Trichomonas hominis (0.88 per cent), Strongyloides stercoralis (0.68 per cent), Hymenolepis nana (0.49 per cent), Chilomastis mesnili (0.10 per cent) and Endolimax nana (0.10 per cent). High incidence of parasitic infections was recorded in females and age group of 6-14 years. PMID- 2559119 TI - Susceptibility status of vectors of Japanese encephalitis to insecticides in south Arcot and Tirunelveli districts of Tamil Nadu. AB - Villages in South Arcot and Tirunelveli districts with high incidence of Japanese Encephalitis were selected for the study of insecticidal resistance to vectors. Culex vishnui and Anopheles hyrcanus group were found to be resistant to DDT (4 per cent for 60 min. exposure) and dieldrin (0.4 per cent for 60 min. exposure) but susceptible to Malathion (5 per cent for 60 min. exposure). So far, this is perhaps the first report of Anopheles hyrcanus group resistant to DDT and dieldrin to be reported from this area. The resistance may be due to extensive use of DDT and HCH for agricultural purposes. PMID- 2559120 TI - Genetic and morphological divergence among sympatric canids. AB - Numerous studies have suggested that the extent of character divergence observed between two sympatric species reflects the intensity of competition for resources or space. However, the influence of time on divergence is often overlooked. We examined the relationship between time and character divergence in two groups of congeneric, sympatric canids on two continents: South American foxes and African jackals. Character divergence was assessed from measurements of body mass and dental and cranial shape. Divergence time was estimated from data on mitochondrial DNA restriction site polymorphisms. Our findings indicate that African jackals are morphologically similar despite having diverged more than 2 million years ago. By contrast, South American foxes differ substantially in both size and morphology after only 250,000 years of evolution. Thus, the lack of character divergence among the African jackals cannot be explained as a result of very recent common ancestry. PMID- 2559121 TI - [A clinicopathological study on cytomegalovirus infection]. AB - A clinicopathological study was carried out in 200 autopsied cases experienced in our department from 1981 to 1988. Cytomegalovirus infection was detected in 18 cases (9.0%). Eleven patients were male and 7 were female, and their ages ranged from 21 to 72 with a mean of 58.1 years. Primary diseases were mainly Non Hodgkin's lymphoma (7 cases) and Adult T-cell leukemia (4 cases), and corticosteroid had been administered to all of them. The most commonly involved organ was lung (77.8%), followed by adrenal (55.6%), esophagus, pancreas, ovary (22.2%), stomach, small intestine, thyroid (16.7%), liver, kidney, tongue (11.1%), and so on. Concomitant infections were frequently complicated, which were bacterial pneumonia (5 cases), fungal pneumonia (3 cases), disseminated varicella-zoster infection (2 cases) and herpes simplex virus esophagitis or stomatitis (5 cases), while, ten patients died of cytomegalovirus pneumonia. Cytomegalovirus infection was one of the fatal opportunistic infections in immunocompromised, especially cell-mediated immunity impaired, hosts such as the patients with lymphocytic malignancies. PMID- 2559122 TI - [Efficacy of ganciclovir against cytomegalovirus nephropathy after kidney transplantation]. AB - A 32-year-old man who had received a kidney transplant from a living related donor, contracted cytomegalovirus (CMV) pneumonitis in the 8th month. He was treated with human interferon-beta and cured of the pneumonitis. After that, his serum creatinine value increased gradually. Renal biopsy revealed the cells with intranuclear inclusion bodies in the renal tubulus and the cells were positive for CMV antigens by direct immunofluorescence test using FITC-labeled mouse monoclonal antibody against an early antigen. He was hospitalized with persistent CMV viruria and treated with ganciclovir. Ganciclovir was administered daily in doses of 3 mg per kg per day for 32 days by intravenous drip infusion and thereafter the same dose was given 3 times weekly for 8 weeks. His urine was positive for CMV before the ganciclovir treatment and became negative on the 31st day after the treatment. The anti-CMV effect of ganciclovir was evidenced by gradual decrease in titer (PFU) of infectious CMV in the urine samples. His serum creatinine value decreased from 3.2 mg/dl to 2.8 mg/dl, and no adverse effect was noticed. Thus, ganciclovir is considered to be efficacious against CMV infections in kidney transplant recipients. PMID- 2559123 TI - [Epidemiology of rotavirus infection around Tokyo area]. AB - Epidemiology of rotavirus infection was studied from 1981 to 1988 mainly in three hospitals around Tokyo area. Major serotypes of rotaviruses in the three places were different from those in two hospitals around Kansai area in Japan (Ref. 6, 13), while, major serotypes were same among three hospitals. Both of serotypes 1 and 4 in group A were mostly found around Tokyo area. Frequencies of type 2, 3, and 9 in group A were low, although the frequencies were various among periods. Detail examinations of rotavirus RNA electropherotypes showed the results as follows; different electropherotypes were found during one winter season and at one hospital, the identical electropherotype was found cross a year and cross a hospital. We could not find the identical electropherotype which belong to two serotypes so far. Seven group C rotaviruses were found since 1987 in three hospitals. It would be important to examine RNA electropherotypes and serotypes for long period not only for epidemiological studies but also for development of vaccine. PMID- 2559124 TI - [Role of nerve growth factor in CNS]. PMID- 2559126 TI - [Regulatory mechanism of the SV40 enhancer activity]. PMID- 2559125 TI - [Transcriptional activation mechanisms of TPA- and cAMP-inducible genes: TRE and CRE]. PMID- 2559127 TI - [NK activity in various liver diseases--especially the relation between NK activity and reserve function of the liver in liver cirrhosis and hepatocellular carcinoma]. AB - NK activity in acute viral hepatitis, chronic hepatitis, liver cirrhosis (LC), metastatic liver cancer and hepatocellular carcinoma (HCC) was preserved and had no significant change compared with the normal control in each disease. In acute hepatitis, NK activity was higher in convalescent phase than in acute phase. Although in LC and HCC, some liver functions such as total bilirubin, the rate of ICG 15' excretion, AFP and IAP did not have significant correlation with NK activity, advanced cases according to Child's classification and E factor which was anatomical extent of HCC in liver showed significantly low NK activity. The low NK activity group advanced much more in Child's classification, performance status, stage and E factor of HCC than high NK activity group significantly. According to those facts, LC or HCC is not a single entity of disease in view point of NK activity. NK activity may change in accordance with the advancement of hepatic disease and reserve function of the liver in LC and HCC. PMID- 2559128 TI - [The clinical studies of intermittent hepatic artery occlusion with infusion chemotherapy--19 cases of unresectable hepatocellular carcinoma and 14 cases of metastatic liver cancer]. AB - For unresectable liver tumors, intermittent occlusion of the feeding artery by means of surgical placement of double lumen balloon catheter in the hepatic artery was combined with chemotherapy. Through the catheter, fluorouracil was continuously infused and one-shot injections of mitomycin or adriamycin were given at the time of balloon inflations. Nineteen patients with primary hepatocellular carcinoma (7 were at Stage III and 11 were at Stage IV) and 14 patients with metastatic liver tumors (4 were at H2 and 9 were at H3 stages) received this newly developed anticancer methodology. In terms of Karnofsky's criteria, 17 of 19 cases with hepatocellular carcinoma achieved Group I responses (89.5%). Fourteen were classified to I-B (73.7%), and one year survival rate was 51.6%. Of 14 cases with metastatic tumors, 13 achieved Group I responses (92.9%). Five were classified to I-B (35.7%), and one year survival was 23.1%. Intermittent occlusion method has advantages in minimizing adverse effects, enabling controlled arterial infusion of anticancer agents and stepwise occlusion of the feeding artery. Despite advanced stages and poor conditions of the patients, remarkable responses were achieved without any serious problems. We are at a conclusion that intermittent occlusion of the hepatic artery when combined with infusion-chemotherapy would, in synergism, be a potential modality against unresectable liver tumors. PMID- 2559129 TI - Clinical effects of cepharanthin (Ceph.) on leukopenia by chemotherapy in lung cancer patients. AB - In order to examine the clinical efficacy of cepharanthin (Ceph.) for preventing leukopenia caused by chemotherapy in lung cancer patients, a randomized trial was carried out. Subjects were 45 patients with non-small cell lung cancer (NSCLC) receiving initial treatment with cisplatin (CDDP) +adriamycin (ADM) +mitomycin C (MMC). Ceph. was given at a dose of 1 mg/kg to the administration group (Ceph. group) and changes in the peripheral leukocyte count were examined in both the Ceph. and non-Ceph. groups. The Ceph. group showed a reduction in the number of cases with a count nadir less than 3,000/microliters (p less than 0.05), a shortening of the time that leukocyte counts stayed under 2,000/microliters, corresponding to the time in which granulocyte counts were less than 500/microliters (p less than 0.05), and also a reduction of the time for leukocyte counts to recover from nadir to 2,000 or 3,000/microliters (p less than 0.05) measured from the commencement of chemotherapy. These results suggest that administration of Ceph. has an antileukopenic effect in anticancer treatment, and makes it possible to reduce the period of risk for infection, allowing effective regimens to be administered repeatedly in shorter intervals. PMID- 2559130 TI - Toxicity of vincristine overdose in a patient with invasive mole. AB - A few reports have documented overdoses of vincristine sulfate. The present report describes our experience with serious complications of a vincristine overdose in an 18-year-old female who had methotrexate-resistant invasive mole. The patient received VAC therapy as the second line chemotherapy after 2 courses of MTX therapy. In the 6th course of VAC therapy, she was given 5 consecutive daily doses of VCR by mistake. On the 5th day of this VAC therapy, she showed the following toxic symptoms: abdominal pain, lumbago, insomnia, bleeding tendency, absence of motor reflex, leukopenia, and paralytic ileus. These symptoms led to realization of the VCR overdose. Leucovorin calcium administration and supportive treatment were carried out. Although it was difficult to evaluate the efficacy of leucovorin calcium on the vincristine toxic symptoms, she recovered and was discharged on the 36th hospital day. PMID- 2559131 TI - [Successful therapy with tegafur and lentinan after TAE for hepatocellular carcinoma--a case report]. AB - A 46-year-old male (performance status 1) with hepatocellular carcinoma (HCC), which diffusely involved the anterior segment of the right lobe, occluded the second portal branch and metastased to the left lobe (stage IV-A), was on combined therapy with TAE of lipiodol (LPD), mitomycin C (MMC) 10 mg and gelatin sponge at the time of angiography on March 16, 1987. Immunochemotherapy with tegafur suppository (Teg sp) 1,000 mg/day and OK432 i.m. was performed for one month after TAE. Instead of OK432, lentinan (LNT) 2 mg in 20% glucose sol. 20 ml i.v./wk was combined with Teg sp 1,000 mg/day from May 13, 1987. Teg sp was reduced to 1,000 mg/2 days from June of the same year, and since then the same therapy of Teg + LNT has been continuing even now. Serum AFP values showing 150 X 10(4) ng/ml at maximum were decreased down to less than 20 ng/ml 13 months after this therapy (total doses: Teg ca. 250 g, LNT 108 mg), remaining still normal. Marked reduction in the size of the lesion (PR) by the therapy has also been proved by the body CT. His PS is now in grade 0. Although in this case the initial steep decline in AFP values was possibly brought about by TAE, the successive antitumor effect indicated by persistent low AFP values may have been mainly due to Teg + LNT. Collection of data on the therapy is needed to clarify this point. PMID- 2559132 TI - [Infectious diseases and monoclonal antibodies]. PMID- 2559133 TI - [A case of Weber-Christian disease with elevated angiotensin-converting enzyme (ACE) activity and myopathy]. PMID- 2559134 TI - Surgical management of large glomus jugulare tumours: infra- and trans-temporal approach. AB - Surgery may have to be considered in the management of large glomus tumours since other modalities of treatment are not curative. The judicious use of surgical intervention is imperative since a neurological deficit involving the last four cranial nerves may ensue and consideration of the quality of life of the patient in the post-operative period is of paramount importance. A brief description of the historical aspects and pathology of these tumours is followed by an analysis of the clinical presentation, investigation and assessment of a series of 10 patients. Six patients underwent surgery and their tumours were removed by the infra- and trans-temporal approach. This is described and illustrated in detail and the surgical results presented. PMID- 2559135 TI - DNA content and proliferative activity of myoepitheliomas. AB - This report adds 16 myoepitheliomas of salivary glands to the 47 already recorded in the literature. It includes, for the first time, a flow cytometric analysis of their ploidy (DNA content) and proliferative capacity (S-phase fraction). Thirteen myoepitheliomas were diploid; three were aneuploid in their DNA content. A high proliferative capacity was always associated with an abnormal DNA content. Only one diploid myoepithelioma had a high S-phase fraction. Both flow-cytometric parameters are good predictors of an aggressive biological behaviour. Recurrences, however, were all the outcome of incomplete primary removal of the myoepitheliomas. Four of the twelve (33 per cent) diploid myoepitheliomas recurred and one, with high S-phase fraction, led to the death of the patient. Two of the three (67 per cent) aneuploid myoepitheliomas recurred. Extensive loco regional invasion by one killed the patient. The other has clinical evidence of distant metastasis. PMID- 2559136 TI - Eicosapentaenoic acid metabolism in brain microvessel endothelium: effect on prostaglandin formation. AB - Mouse brain microvessel endothelial cells convert eicosapentaenoic acid (EPA) to prostaglandin (PG) E3, PGI3, and several hydroxy fatty acid derivatives. Similar types of products are formed by these microvessel endothelial cells from arachidonic acid. The formation of PGI2 and PGE2 is reduced, however, when the brain microvessel endothelial cultures are incubated initially with EPA. Exposure to linolenic or docosahexaenoic acid also decreased the capacity of these microvessel endothelial cells to form PGI2 and PGE2, but the reductions were smaller than those produced by EPA. Like the endothelial cultures, intact mouse brain microvessels convert EPA into eicosanoids, and incubation with EPA reduces the subsequent capacity of the microvessels to produce PGI2 and PGE2. Brain microvessel endothelial cells took up less EPA than arachidonic acid, primarily due to lesser incorporation into the inositol, ethanolamine, and serine glycerophospholipids. By contrast, considerably more EPA than arachidonic acid was incorporated into triglycerides. These findings suggest that the microvessel endothelium may be a site of conversion of EPA to eicosanoids in the brain and that EPA availability can influence the amount of dienoic prostaglandins released by the brain microvasculature. Furthermore, the substantial incorporation of EPA into triglyceride suggests that this neutral lipid may play an important role in the processing and metabolism of EPA in brain microvessels. PMID- 2559137 TI - Functional size of acyl coenzyme A:diacylglycerol acyltransferase by radiation inactivation. AB - Rat liver acyl coenzyme A:diacylglycerol acyltransferase, an intrinsic membrane activity associated with the endoplasmic reticulum, catalyzes the terminal and rate-limiting step in triglyceride synthesis. This enzyme has never been purified nor has its gene been isolated. Inactivation by ionizing radiation and target analysis were used to determine its functional size in situ. Monoexponential radiation inactivation curves were obtained which indicated that a single-sized unit of 72 +/- 4 kDa is required for expression of activity. The size corresponds only to the protein portion of the target and may represent one or several polypeptides. PMID- 2559138 TI - Actions of dendrotoxin on K+ channels and neuromuscular transmission in Drosophila melanogaster, and its effects in synergy with K+ channel-specific drugs and mutations. AB - The blockade of K+ channels and enhancement of neuromuscular transmission by dendrotoxin (DTX), a convulsant peptide from mamba snake venom, were examined in normal and mutant larval preparations of Drosophila. Two-microelectrode voltage clamp experiments showed that DTX reduced the transient K+ current, IA, in muscle membrane. This effect was suppressed by raising the Mg2+ concentration or by lowering the temperature. The interaction of DTX with Mg2+ was further analyzed at a low cation concentration, at which DTX reduced both IA and the delayed rectifier IK. These results were correlated with the action of DTX on the neuromuscular junction. Its facilitatory effect on excitatory junctional potentials (EJPs) was relatively mild but the effect was drastically enhanced when combined with certain mutations and K+ channel blocking drugs, leading to repetitive or prolonged giant EJPs. Only the mutations or drugs that reduced IK or the Ca2(+)-dependent K+ current, ICF, could yield these synergistic effects with DTX. In contrast, the abnormal EJPs caused by the mutation or drug that blocked IA were not further enhanced by DTX, indicating that DTX also affects IA at the neuromuscular junction. Thus, the A-type K+ channels in muscle and nerve terminals appeared very similar in their sensitivity to the specific toxin, drugs and mutations examined here. PMID- 2559139 TI - Nimodipine block of calcium channels in rat vascular smooth muscle cell lines. Exceptionally high-affinity binding in A7r5 and A10 cells. AB - Calcium channel currents were studied in the A10 and A7r5 cell lines derived from rat thoracic aorta muscle cells. The whole-cell variation of the patch voltage clamp technique was used. Results with each cell line were nearly identical. Two types of Ca channels were found in each cell line that are similar to the L-type and T-type Ca channels found in excitable cells. Nimodipine block of the L-type Ca channels in both cell lines is more potent than in previously studied tissues. The kinetics of nimodipine block are accounted for by a model that postulates 1:1 drug binding to open Ca channels with an apparent dissociation constant (KO) of 16-45 pM. In A7r5 cells, the rate of onset of nimodipine block increases with the test potential, in quantitative agreement with the model of open channel block. The apparent association rate (f) is 1.4 x 10(9) M-1 s-1; the dissociation rate (b) is about 0.024 s-1. In anterior pituitary cells (GH4C1 cells), KO is 30 times larger; b is only twice as fast, but f is 15 times slower. The comparative kinetic analysis indicates that the high-affinity binding site for nimodipine is similar in both GH4C1 and A7r5 cells, but nimodipine diffuses much faster or has a larger partition coefficient into the plasmalemma of A7r5 cells than for GH4C1 cells. Unusually high-affinity binding was not observed in earlier 45Ca flux studies with A10 and A7r5 cells. The model of open channel block accounts for the discrepancy; only a small fraction of the Ca channels are in the high affinity open state under the conditions used in 45Ca flux studies, so an effective binding constant is measured that is much greater than the dissociation constant for high-affinity binding. PMID- 2559143 TI - Bacterial ethylene synthesis from 2-oxo-4-thiobutyric acid and from methionine. AB - The ability of selected bacterial cultures to synthesize ethylene during growth in nutrient broth supplemented with methionine or 2-oxo-4-methylthiobutyric acid (KMBA) was examined. Although most cultures transformed KMBA into ethylene, only those of Escherichia coli SPAO and Chromobacterium violaceum were able to convert exogenously added methionine to ethylene. In chemically defined media, E. coli SPAO produced the highest amounts of ethylene from methionine and KMBA. This capability was affected by the nature of the carbon source and the type and amount of nitrogen source used for growth. When glutamate was used as sole source of carbon and nitrogen for growth, the activity of the ethylenogenic enzymes was reduced to 25% of that observed with cultures grown with glucose and NH4Cl. Neither methionine nor KMBA significantly affected the ethylenogenic capacity of E. coli SPAO. Menadione and paraquat, compounds that generate superoxide radicals, stimulated ethylene synthesis by harvested cells, but not by cell-free extracts of E. coli SPAO. In addition, cells of Pseudomonas aeruginosa, which produced no ethylene in culture in the presence of exogenously added KMBA, yet possessed the necessary enzymes in an active form, were able to synthesize ethylene from KMBA when incubated with menadione or paraquat. PMID- 2559141 TI - Kinetics of contraction initiated by flash photolysis of caged adenosine triphosphate in tonic and phasic smooth muscles. AB - Laser flash photolysis of caged adenosine triphosphate (ATP), in the presence of Ca2+, was used to examine the time course of isometric force development from rigor states in glycerinated tonic (rabbit trachealis) and phasic (guinea-pig ileum and portal vein) smooth muscles. Photolytic liberation of ATP from caged ATP initiated force development, at 20 degrees C, with half-time (t1/2) of 5.4 s in trachealis and 1.2-2.2 s in the phasic muscles. Prior to photolysis, some muscles were phosphorylated with ATP plus okadaic acid (an inhibitor of myosin light-chain phosphatase) or thiophosphorylated with ATP gamma S to fully activate the regulatory system, before turning on the contractile apparatus. In these prephosphorylated muscles, force development, after caged ATP photolysis, was more rapid than in the unphosphorylated muscles, but the t1/2 values for trachealis (0.8-1.1 s) were still longer than for ileum and portal-vein muscles (0.20-0.25 s). The results suggest that both the contractile machinery and the regulatory system are slower in the tonic than in the phasic smooth muscles. The time course of force development for each muscle type was sigmoidal, with an initial delay (td) of approximately 10% of the t1/2 value. Some possible chemical and mechanical origins of the delay are discussed. PMID- 2559140 TI - Effects of magnesium on inactivation of the voltage-gated calcium current in cardiac myocytes. AB - The effects of changes in intracellular and extracellular free ionized [Mg2+] on inactivation of ICa and IBa in isolated ventricular myocytes of the frog were investigated using the whole-cell configuration of the patch-clamp technique. Intracellular [Mg2+] was varied by internal perfusion with solutions having different calculated free [Mg2+]. Increasing [Mg2+]i from 0.3 mM to 3.0 mM caused a 16% reduction in peak ICa amplitude and a 36% reduction in peak IBa amplitude, shifted the current-voltage relationship and the inactivation curve approximately 10 mV to the left, decreased relief from inactivation, and caused a dramatic increase in the rate of inactivation of IBa. The shifts in the current-voltage and inactivation curves were attributed to screening of internal surface charge by Mg2+. The increased rate of inactivation of IBa was due to an increase in both the steady-state level of inactivation as well as an increase in the rate of inactivation, as measured by two-pulse inactivation protocols. Increasing external [Mg2+] decreased IBa amplitude and shifted the current-voltage and inactivation curves to the right, but, in contrast to the effect of internal Mg2+, had little effect on the inactivation kinetics or the steady-state inactivation of IBa at potentials positive to 0 mV. These observations suggest that the Ca channel can be blocked quite rapidly by external Mg2+, whereas the block by [Mg2+]i is time and voltage dependent. We propose that inactivation of Ca channels can occur by both calcium-dependent and purely voltage-dependent mechanisms, and that a component of voltage-dependent inactivation can be modulated by changes in cytoplasmic Mg2+. PMID- 2559142 TI - Activation of three types of membrane currents by various divalent cations in identified molluscan pacemaker neurons. AB - We investigated membrane currents activated by intracellular divalent cations in two types of molluscan pacemaker neurons. A fast and quantitative pressure injection technique was used to apply Ca2+ and other divalent cations. Ca2+ was most effective in activating a nonspecific cation current and two types of K+ currents found in these cells. One type of outward current was quickly activated following injections with increasing effectiveness for divalent cations of ionic radii that were closer to the radius of Ca2+ (Ca2+ greater than Cd2+ greater than Hg2+ greater than Mn2+ greater than Zn2+ greater than Co2+ greater than Ni2+ greater than Pb2+ greater than Sr2+ greater than Mg2+ greater than Ba2+). The other type of outward current was activated with a delay by Ca2+ greater than Sr2+ greater than Hg2+ greater than Pb2+. Mg2+, Ba2+, Zn2+, Cd2+, Mn2+, Co2+, and Ni2+ were ineffective in concentrations up to 5 mM. Comparison with properties of Ca2(+)-sensitive proteins related to the binding of divalent cations suggests that a Ca2(+)-binding protein of the calmodulin/troponin C type is involved in Ca2(+)-dependent activation of the fast-activated type of K+ current. Th sequence obtained for the slowly activated type is compatible with the effectiveness of different divalent cations in activating protein kinase C. The nonspecific cation current was activated by Ca2+ greater than Hg2+ greater than Ba2+ greater than Pb2+ greater than Sr2+, a sequence unlike sequences for known Ca2(+)-binding proteins. PMID- 2559144 TI - Characterization of Pseudomonas mercury-resistance transposon Tn502, which has a preferred insertion site in RP1. AB - Tn502mer differs in size and restriction map from the well-characterized Tn501mer. It also differs in its preferential and high-frequency insertion into the 6 kb PstI-C region of RP1. The affinity for this region is perpetuated in pVS76, a clone of RP1 PstI-C in pBR322. Restriction mapping of independent pVS76::Tn502 derivatives revealed that Tn502 inserted at the same site (or small region) in PstI-C corresponding to the 35 kb coordinate in RP1. Insertion occurred in both orientations, but one was preferred. When PstI-C was deleted from RP1, acquisition of Tn502 was reduced and the sites of insertion randomized. PMID- 2559145 TI - Chromosomal location of the Bacillus subtilis aspartokinase II gene and nucleotide sequence of the adjacent genes homologous to uvrC and trx of Escherichia coli. AB - The aspartokinase II (ask) operon of Bacillus subtilis consists of two in-phase overlapping genes that encode the two subunits of the lysine-sensitive isoenzyme of aspartokinase (ATP:L-aspartate 4-phosphotransferase, EC 2.7.2.4). Transduction mapping of the ask operon, inactivated by recombinational insertion of a cat marker, indicates a chromosomal location (about 253 degrees) between leuA and aroG. ask is thus remote from aecB, eliminating aecB as a possible locus for the structural gene of aspartokinase II, but close to aecA and uvrB. The nucleotide sequence of a 2 kb DNA fragment just upstream of the ask operon was determined and found to contain two open reading frames. The deduced amino acid sequence of the distal reading frame exhibits extensive homology with Escherichia coli thioredoxin and that of the proximal one, which overlaps with the ask promoter, is homologous to the deduced product of the E. coli uvrC gene. Insertional mutagenesis of the proximal open reading frame led to a mitomycin-sensitive phenotype, consistent with a role in DNA repair. In conjunction with the data of M. Petricek, L. Rutberg & L. Hederstedt [FEMS Microbiology Letters 61, 85-88] our results define the nucleotide sequence of an 8.8 kb segment of the B. subtilis chromosome near 253 degrees and the following order of genes: trx-uvrB-ask-orfX sdhC-sdhA-sdhB-orfY++ +-gerE. PMID- 2559146 TI - Transposon-916-like elements in clinical isolates of Enterococcus faecium. AB - Tetracycline (Tc) resistance was found in nine out of ten clinical isolates of Enterococcus faecium. Conjugative transposons, designated Tn5031, Tn5032 and Tn5033, were present in the chromosome of three isolates. The transposons were similar both structurally and functionally to Tn916 containing the tetM determinant. A large non-conjugative plasmid found in a fourth isolate contained an element homologous to Tn916. The four isolates containing the element showing homology to Tn916 exhibited a substantially higher level of Tc resistance than the remaining five Tc-resistant isolates. Tc-resistance genes which have not been identified are apparently responsible for the low-level Tc resistance in five clinical isolates. PMID- 2559147 TI - Molecular cloning and expression in Escherichia coli of the recA gene of Legionella pneumophila. AB - Interspecific complementation of an Escherichia coli recA mutant with a Legionella pneumophila genomic library was used to identify a recombinant plasmid encoding the L. pneumophila recA gene. Recombinant E. coli strains harbouring the L. pneumophila recA gene were isolated by replica-plating bacterial colonies on medium containing methyl methanesulphonate (MMS). MMS-resistant clones were identified as encoding the L. pneumophila recA analogue by their ability to protect E. coli HB101 from UV exposure and promote homologous recombination. Subcloning of selected restriction fragments and Tn5 mutagenesis localized the recA gene to a 1.7 kb Bg/II-EcoRI fragment. Analysis of minicell preparations harbouring a 1.9 kb EcoRI fragment containing the recA coding segment revealed a single 37.5 kDa protein. Insertional inactivation of the cloned recA gene by Tn5 resulted in the disappearance of the 37.5 kDa protein, concomitant with the loss of RecA function. The L. pneumophila recA gene product did not promote induction of a lambda lysogen; instead, the presence of the heterologous recA gene caused a significant reduction in spontaneous and mitomycin-C-induced prophage induction in recA+ and recA E. coli backgrounds. Despite the lack of significant genetic homology between the L. pneumophila recA gene and the E. coli counterpart, the L. pneumophila RecA protein was nearly identical to that of E. coli in molecular mass, and the two proteins showed antigenic cross-reactivity. Western blot analysis of UV-treated L. pneumophila revealed a significant increase in RecA antigen in irradiated versus control cells, suggesting that the L. pneumophila recA gene is regulated in a manner similar to that of E. coli recA. PMID- 2559148 TI - Coat and enterotoxin-related proteins in Clostridium perfringens spores. AB - Coat proteins from mature spores of two enterotoxin-positive (Ent+) and two enterotoxin-negative (Ent-) strains of Clostridium perfringens were solubilized using 50 mM-dithiothreitol and 1% sodium dodecyl sulphate at pH 9.7, and alkylated using 110 mM-iodoacetamide to prevent aggregation. The coat proteins and C. perfringens type A enterotoxin (CPE) were separated by SDS-PAGE and analysed by Western blotting using anti-CPE antibody. As previously reported, CPE aggregated in the presence of SDS, but no aggregation occurred at concentrations below 15 micrograms CPE ml-1. Two CPE-related proteins (34 and 48 kDa) were found in the solubilized spore coat protein of Ent+ strains while only the 48 kDa CPE related protein was found in the spore coat fraction of Ent- strains. CPE-related proteins comprised 2.7% and 0.8% of the total solubilized coat protein of Ent+ and Ent- strains respectively. CPE-related proteins could be extracted from the spores with 1% SDS alone. They could also be released by disruption of whole spores, indicating that the CPE-related proteins may be in the spore core or trapped between the core and coat layers. The results suggest that CPE is not a major structural component of the coat fraction of C. perfringens spores. PMID- 2559149 TI - Antibiotic susceptibility of Salmonella spp. at different pH values. AB - We have examined the effects of acidic pH, in the range of those prevailing within phagosomes and lysosomes, on the growth and the susceptibility to different antibiotics of several strains of Salmonella spp. The minimal inhibitory concentration and the minimal bactericidal concentration of several beta-lactams were increased considerably during culture at pH 5.2. The extent of the increase was a function of: (1) the beta-lactam structure and, more particularly, the hydrophobicity of the side-chain of the molecule; and (2) the bacterial serotype. This phenotypic resistance at acid pH was not due to beta lactamase activity or to a lower growth rate. In contrast, rifamycin SV was more active at acidic pH than at neutral pH and chloramphenicol, another highly hydrophobic drug, was equally efficacious at both pH values. Membrane lipopolysaccharide mutants, but not porin mutants, cultivated at an acidic pH were inhibited by lower concentrations of the beta-lactams. This suggests that the increased resistance to beta-lactams, and the increased susceptibility to rifamycin SV, at acidic pH, could have resulted from modified permeability of the outer membrane to antibiotics. PMID- 2559150 TI - cAMP- and RAS-independent nutritional regulation of plasma-membrane H+-ATPase activity in Saccharomyces cerevisiae. AB - The plasma-membrane ATPase of Saccharomyces cerevisiae is a proton pump whose activity, essential fro proliferation, is subject to regulation by nutritional signals. The previous finding that the CDC25 gene product is required for the glucose-induced H+-ATPase activation suggested that H+-ATPase activity is regulated by cAMP. Analysis of starvation-induced inactivation and glucose induced activation of the H+-ATPase in mutants affected in activity of the RAS proteins, adenylyl cyclase or cAMP-dependent protein kinase showed that nutritional regulation of H+-ATPase activity does not depend directly on any of these factors. We conclude that adenlyl cyclase does not mediate all nutritional responses. This also indicates that the specific CDC25 requirement for the glucose-induced activation of the H+-ATPase identifies a new function for the CDC25 gene product, a function that appears to be independent of CDC25-mediated modulation of the RAS/adenylyl cyclase/cAMP pathway. PMID- 2559151 TI - Analysis of separate isolates of Bordetella pertussis repeated DNA sequences. AB - Two independent isolates of a Bordetella pertussis repeated DNA unit were sequenced and shown to be an insertion sequence element with five nucleotide differences between the two copies. The sequences were 1053 bp in length with near-perfect terminal inverted repeats of 28 bp, had three open reading frames, and were each flanked by short direct repeats. The two insertion sequences showed considerable homology to two other B. pertussis repeated DNA sequences reported recently: IS481 and a 530 bp repeated DNA unit. The B. pertussis insertion sequence would appear to comprise a group of closely related sequences differing mainly in flanking direct repeats and the terminal inverted repeats. The two isolates reported here, which were from the adenylate cyclase and agglutinogen 2 regions of the genome, were numbered IS48lvl and IS48lv2 respectively. PMID- 2559152 TI - Inhibition of growth of Chlamydia trachomatis by the calcium antagonist verapamil. AB - Treatment of BGM (African Green Monkey kidney) cells with the calcium antagonist Verapamil resulted in a reduced yield of chlamydial infectious particles. The inhibitory effect was concentration-dependent, the maximal effect being achieved at 200 microM-Verapamil, which produced a 99.99% reduction of infectious particle yield. Electron microscopy showed that control Chlamydia trachomatis-infected BGM cells contained typical large inclusions in which most of the particles were elementary bodies, whereas Verapamil-treated infected cells contained small inclusions consisting predominantly of reticulate bodies. The findings indicate a possible therapeutic use of this calcium antagonist as an anti-chlamydial drug. PMID- 2559153 TI - Mutations affecting the cytochrome d-containing oxidase complex of Escherichia coli K12: identification and mapping of a fourth locus, cydD. AB - A mutant of Escherichia coli K12 has been isolated affected in a gene, designated cydD, distinct from the three previously described loci involved in the synthesis of assembly of the cytochrome bd oxidase complex. The mutant, obtained by nitrosoguanidine mutagenesis, lacks the spectroscopically detectable components of this oxidase, namely cytochromes b558, b595 and d. Cytochrome oxidase o is the sole CO-binding cytochrome in membranes of the mutant, but the soluble haemoprotein b-590 and catalase activity appear unaffected. Discrimination between Cyd+ and Cyd- strains is facilitated by the development of a defined low phosphate medium that allows the inclusion of Zn2+ as well as azide, inhibitors of respiratory electron transfer particularly via cytochrome o. Mapping with F prime factors and by P1 cotransductional frequencies shows the mutation to map near 19.3 min on the E. coli chromosome, distinct from cydC, which maps at 18.9 min. The gene order in this region was tested in a three-factor cross and demonstrates the order zbj::Tn10(YYC199)-cydD-aroA, consistent with cotransduction frequencies. PMID- 2559154 TI - Interaction of Candida albicans with neutrophils: effect of phenotypic changes in yeast cell-surface composition. AB - The susceptibility of four strains of Candida albicans to phagocytosis and intracellular killing by rabbit peritoneal neutrophils was investigated. Two of the strains, isolated from active infections, were known to synthesize a surface layer of mannoprotein fibrils in response to growth on 500 mm-galactose; the other strains, from asymptomatic carriers, lacked this capability. The presence of serum opsonins greatly enhanced phagocytosis of all four strains and, following opsonization, phagocytosis of an infective strain was equally rapid after growth on either 500 mm-galactose or 50 mm-glucose. In the absence of opsonins, galactose-grown infective strains were phagocytosed faster than either glucose-grown infective strains or galactose-grown carrier strains. These differences in phagocytic uptake were paralleled by differences in neutrophil chemiluminescence response. Intracellular killing of galactose-grown infective strains was only half that of glucose-grown infective strains or galactose-grown carrier strains after incubation for 60 min. Pretreatment of neutrophils with extracellular polymeric material, which contains the surface fibrils, completely inhibited intracellular killing. These results indicate that production of the fibrillar layer promotes yeast virulence by increasing resistance to intracellular killing, although it may enhance phagocytosis in locations where opsonic activity is poor. PMID- 2559155 TI - Steroid catechol degradation: disecoandrostane intermediates accumulated by Pseudomonas transposon mutant strains. AB - Eleven transposon mutant strains affected in bile acid catabolism were each found to form yellow, muconic-like intermediates from bile acids. To characterize these unstable intermediates, media from the growth of one of these mutants with deoxycholic acid was treated with ammonia, then the crude product was methylated with diazomethane. Four compounds were subsequently isolated; spectral evidence suggested that they were methyl 12 alpha-hydroxy-3-oxo-23,24-dinorchola-1,4-dien 22-oate, methyl 4-aza-12 beta-hydroxy-9(10)-secoandrosta-1,3,5-triene-9,17-dione 3-carboxyl ate, 4-aza-9 alpha, 12 beta-dihydroxy-9(10)-secoandrosta-1,3,5-trien 17-one-3- methyl carboxylate and 4 alpha-[3'-propionic acid]-5-amino-7 beta hydroxy-7 alpha beta-methyl- 3a alpha, 4,7,7a-tetrahydro-1-indanone-delta-lactam. It is proposed that the mutants are blocked in the utilization of such muconic like compounds as the 3,12 beta-dihydroxy-5,9,17-trioxo-4(5),9(10)- disecoandrostal (10),2-dien-4-oic acid formed from deoxycholic acid. A further mutant was examined, which converted deoxycholic acid to 12 alpha-hydroxyandrosta 1,4-dien-3,17-dione, but accumulated yellow products from steroids which lacked a 12 alpha-hydroxy function, such as chenodeoxycholic acid. The products from the latter acid were treated as above; spectral evidence suggested that the two compounds isolated were methyl 4-aza-7-hydroxy-9(10)-secoandrosta-1,3,5- triene 9,17-dione-3-carboxylate and 4 alpha-[1'alpha-hydroxy-3'-propionic acid]-5-amino 7a beta-methyl-3a alpha,4,7,7a-tetrahydro-1-indanone-delta-lactam. PMID- 2559156 TI - The genetics of bile acid degradation in Pseudomonas spp.: location and cloning of catabolic genes. AB - Four Pseudomonas spp. capable of utilizing bile acids as sole carbon source were examined for the presence of plasmids. One plasmid was found in Pseudomonas sp. RAL8, but no plasmids could be detected in the other three strains. Mitomycin C curing of RAL8 did not affect the ability of the strain to grow on bile acids. This suggested that the genetic information for bile acid catabolism in all four strains was chromosomally located. To isolate bile acid catabolic genes. DNA from RAL8 was partially digested with Sau3A, then the DNA fragments cloned into the broad-host-range cosmid vector pMMB33. The resulting gene bank was screened by plate-mating with two stable RAL8 mutants. Four of the gene bank clones were found to give a positive complementation with one or both mutants. Examination of the plasmids in the four clones revealed that they were unstable, but detailed mapping enabled a 52 kb restriction map to be derived. Further complementation work showed that two of the bile acid catabolic genes are located close together on the map, and may be contiguous. PMID- 2559157 TI - Seroepidemiology of hepatitis A, B, and D viruses and human T-lymphocyte tropic viruses in Japanese drug abusers. AB - To evaluate the prevalence of hepatitis virus markers and human T-cell lymphotropic virus infections among drug abusers in Japan, serum samples were collected from 91 male drug abusers at the Shinshu University Hospital and the rehabilitation facility in Matsumoto and from 519 healthy male blood donors as controls. Sera were tested for antibody to hepatitis A virus (anti-HAV), hepatitis B surface antigen (HBsAg), antibody to HBsAg (anti-HBs), antibody to hepatitis B core antigen (anti-HBc), immunoglobulin M anti-HBc (IgM anti-HBc), antibody to hepatitis D virus (anti-HDV), antibody to HTLV type 1 (anti-HTLV 1), and antibody to human immunodeficiency virus (anti-HIV). The prevalence of anti HAV was 13.2% in drug abusers and 10.8% in controls (not significant). The prevalences of HBsAg, anti-HBs, anti-HBc and exposure rate to hepatitis B virus (HBV) were 4.4%, 24.2%, 31.9%, and 35.2%, respectively, in drug abusers and 0.8%, 6.7%, 9.6%, and 9.6% in controls. The exposure rate to HBV was significantly different (P less than 0.001). IgM anti-HBc and anti-HDV were not detected in any sera. Anti-HTLV I was detected in three drug abusers (3.3%) and in one (0.2%) of the controls (P less than 0.01). All sera were negative for anti-HIV in all subjects. Infection with HBV and HTLV I is more common among drug abusers than in the general population of blood donors in Japan. PMID- 2559158 TI - Formalin inactivated Junin virus: immunogenicity and protection assays. AB - The aim of this study was to determine if Junin virus inactivated with formalin (FA) was immunogenic and able to elicit a protective response in the guinea pig. The XJ-Clone 3 strain of Junin virus grown in Vero cells was exposed to FA at 0 degrees C. The following inactivated antigens were prepared: A1, 0.1% FA for 50 hr; A2, 0.1% FA for 50 hr followed by concentration with polyethylene glycol (PEG); B1, 0.05% FA for 70 hr; B2, 0.05% FA for 70 hr plus PEG concentration; C, 0.1% FA for 50 hr followed by ultracentrifugation and purification by sucrose gradient. No residual infectivity was detected in any inactivated antigen either after two passages in newborn mice or by coculture with Vero cells. After immunization, high-neutralizing and low-immunofluorescent antibody titers wer obtained in adult mice and guinea pigs, thus showing that antigenicity was preserved. However, in spite of the presence of neutralizing antibodies, guinea pigs gained no protection against challenge with the highly pathogenic XJ strain. These results suggest that antibody amount and/or quality may be inadequate; alternatively, mechanisms other than humoral immunity, such as cellular response, not elicited by inactivated antigens may be essential for protection against Junin virus. PMID- 2559159 TI - Laboratory diagnosis of Japanese encephalitis using monoclonal antibodies and correlation of findings with the outcome. AB - Detection of virus, viral antigen, and class-specific antibody was carried out in cerebrospinal fluid (CSF) and sera of 27 children with Japanese encephalitis. The diagnosis could be confirmed in 78.57% of cases (22/27) by demonstration of virus specific IgM in CSF (15/22), viral antigen in CSF (5/22), or by virus isolation (2/22). Absence of virus specific IgM in CSF was associated with a fatal outcome (P = 0.05). PMID- 2559160 TI - Herpes viruses and duodenal ulcer disease. AB - Antibodies to herpes simplex virus and Epstein-Barr virus were sought in the sera of 33 patients with endoscopically proved duodenal ulcer and 33 healthy controls. Neither the prevalence nor the antibody levels to both viruses were significantly different between the two groups. These results suggest that herpes viruses may not play a role in the pathogenesis of duodenal ulcer disease. PMID- 2559161 TI - The spectrum of polyneuropathies in patients infected with HIV. AB - Twenty five patients with peripheral neuropathy at different stages of human immunodeficiency virus (HIV) infection are reported. Cerebrospinal fluid (CSF) findings were available in 17 cases, electrophysiology in all and a neuromuscular biopsy in 11. Of six otherwise asymptomatic HIV+ patients, five had chronic inflammatory demyelinating polyneuropathy (CIDP) and one acute inflammatory demyelinating polyneuropathy (AIDP). CSF showed pleocytosis in all cases. Infiltration of the endoneurium and/or the epineurium by mononuclear cells was seen in biopsies from three cases. These six patients recovered either spontaneously, or with corticosteroids or plasmaphereses. Of five patients with AIDS related complex (ARC), three had distal predominantly sensory peripheral neuropathy (DSPN), one CIDP and one mixed neuropathy. Of 14 patients with AIDS, one had mononeuropathy multiplex and 13 painful DSPN. Electrophysiological studies were consistent with an axonopathy. Nerve biopsies in six cases showed axonal changes but surprisingly associated with marked segmental demyelination in two cases. Cell infiltration was present in nerve samples in two cases. Five patients died within six months after the onset of the neuropathy. PMID- 2559163 TI - Amyloidosis of the nervous system. AB - Various types of amyloid fibril deposits occur in the nervous system with unique clinical characteristics and pathogeneses. Genetic mutations cause the familial amyloidotic polyneuropathies and acquired polyneuropathies occurring particularly in patients suffering from hypernephromas and myelomas also result from the production of abnormal proteins. Amyloid fibril deposits in cerebral plaques and vessels consisting of beta-protein are seen in acquired and familial Alzheimer's disease and in Down's syndrome individuals over 40 years of age. This amyloid fibril deposition could result from a mutational, transcriptional or post translational alteration in these pathologic processes with most evidence supporting the latter. Other diseases including hereditary cerebral hemorrhage of the Dutch type and Batten's disease involve beta-amyloid deposition. The features of the familial and transmissible forms of the spongiform encephalopathies are associated with the prion protein which comprises the amyloid fibril deposits in these conditions. This wide variety of nervous system disorders having amyloid deposits as their primary or subsidiary characteristic make studies of these conditions intriguing models for research workers in clinical, pathologic and molecular biologic fields. PMID- 2559162 TI - Hypothyroidism and polyneuropathy. AB - The prevalence and characteristics of polyneuropathy were assessed using standard clinical and electrophysiological criteria in 39 consecutive outpatients with primary hypothyroidism, 15 of whom were previously untreated. Subjective complaints, mainly paraesthesiae, were recorded from 25 cases (64%) and objective findings supporting a clinical diagnosis of polyneuropathy were present in 13 (33%). Using standard electrophysiological criteria, a definite diagnosis of polyneuropathy was made in 28 cases (72%). The commonest sites of abnormal nerve conduction were the sensory nerves, especially the sural nerve. Polyneuropathy was generally mild. None of the clinical and biochemical indicators of hypothyroidism were significantly correlated with the electrophysiological signs of peripheral nerve impairment or the diagnosis of polyneuropathy. PMID- 2559164 TI - Effects of different opiates on hypothalamic monoamine turnover and on plasma LH levels in pro-oestrous rats. AB - Opiate inhibition of luteinizing hormone (LH) appears to involve changes in hypothalamic monoaminergic activity. Agonists of mu-, kappa- and sigma-opioid receptors and an opiate antagonist were administered at the onset of the preovulatory LH surge and their effects on hypothalamic monoamine turnover, and on plasma LH levels, investigated. The opiate antagonist, naloxone, significantly increased both noradrenaline (NA) turnover and plasma LH levels. Morphine (mu agonist), significantly decreased NA concentration and plasma LH levels, but significantly increased dopamine (DA) and serotonin (5-HT) activity. Levorphanol (another mu-agonist) significantly decreased both NA and 5-HT concentrations and had no effect on circulating LH. Cyclazocine, ketocyclazocine and tifluadom (kappa-agonists) increased NA turnover but only tifluadom increased 5-HT turnover, also reducing LH levels significantly. N-Allylnormetazocine (SKF 10,047; sigma-agonist) increased 5-HT activity but did not alter LH levels. This study has confirmed the existence of a heterogenous group of opioid receptors within the hypothalamus which modulate monoamine neurotransmitters controlling LH release. PMID- 2559165 TI - Glial cytoplasmic inclusions in the CNS of patients with multiple system atrophy (striatonigral degeneration, olivopontocerebellar atrophy and Shy-Drager syndrome). AB - Glial cytoplasmic inclusions (GCIs) were demonstrated by silver staining, immunocytochemistry and by electron microscopy in the central nervous system (CNS) of 11 patients with various combinations of striatonigral degeneration, olivopontocerebellar atrophy and Shy-Drager syndrome. Although their configuration in light microscope can sometimes resemble neurofibrillary tangles, their cellular localisation, measurements, ultrastructure, immunocytochemical characteristics and regional distribution all differ from these Alzheimer type changes. The majority of GCIs were localized in the white matter and appeared to be accompanied by an increase in the number of interfascicular oligodendroglial cells and pallor or loss of myelin staining. Our histological, ultrastructural and immunocytochemical findings all indicate that the cells which contain GCIs are oligodendrocytes and the inclusions themselves are composed of tubular structures. The presence of the until now unknown GCIs in all the 11 CNS, but not in age- and sex-matched control brains, indicates that GCI is a cellular change characteristic of multiple system atrophy and the three syndromes are various manifestations of the same disease. PMID- 2559166 TI - Histochemical characterization of sensory neurons with high-affinity receptors for nerve growth factor. AB - Approximately one half of the neurons in the lumbar dorsal root ganglion of adult rats display high-affinity receptors for nerve growth factor (NGF). To ascertain which types of sensory neurons are potentially responsive to NGF, adjacent cryostat sections of rat dorsal root ganglia were processed either for NGF receptor using radioautography or by one of four histochemical procedures. Histograms of the densities of neuronal labelling by radioiodinated NGF were examined for subpopulations of lumbar sensory neurons with thiamine monophosphatase enzyme activity or with immunoreactivity for calcitonin gene related peptide (CGRP), substance P, or somatostatin. Virtually all neurons with strong CGRP immunoreactivity had high-affinity NGF binding sites, although some neurons with faintly positive CGRP immunoreactivity lacked such NGF binding. A subpopulation of large neurons, approximately 5% of the total, had dense labelling by 125I-NGF but were not stained by this immunohistochemical technique for CGRP. Of the three major populations of small neurons those with substance P immunoreactivity were consistently and heavily labelled by radioiodinated NGF whereas those with somatostatin immunoreactivity or thiamine monophosphatase activity were not specifically labelled by radioautography. For these primary sensory neurons in mature rats the genes for substance P and CGRP seem to be strongly expressed only in neurons capable of responding to NGF. On the other hand, neurons containing somatostatin and thiamine monophosphatase invariably lack high-affinity NGF receptors. PMID- 2559167 TI - Rapid quantitative immunohistochemical assessment of human peripheral neuropathies using a monoclonal antibody against nerve growth factor receptor. AB - An analysis of nerve growth factor (NGF) receptor expression and density in human sural nerve biopsies was performed by immunocytochemistry with a murine monoclonal antibody against the human NGF receptor. Quantitative assessment of immunostaining density was made by histospectrophotometry on frozen sections. Although there was enhanced expression of NGF receptor within endoneurium in all patients with clinical neuropathies, expression was highest in nerves with axonal disease, consistent with the proposal that disruption of axon-Schwann cell interactions triggers the re-expression of the NGF receptor. These results with human nerves, together with previous studies with animal models, suggest that NGF and NGF receptor play important roles in the general response to neuronal injury. PMID- 2559168 TI - C-type virus particles in placentas of rhesus monkeys after maternal treatment with recombinant leukocyte A interferon. AB - Interferon inhibits the normal replication of C-type virus particles grown in vitro. The ability of interferon to produce a similar effect in vivo was tested in rhesus monkey placentas which are known to contain C-type virus particles. Samples of placenta from 6 monkeys treated with recombinant leukocyte A interferon (25 x 10(6) units/kg) for 7 to 21 days and from 6 control monkeys were examined in the electron microscope. Budding and immature C-type virus particles were present in every placenta examined from both interferon-treated and control monkeys. Mature particles were found in placentas from 3 of 6 monkeys in both treated and control groups. Possible explanations for the absence of detectable effects of interferon in C-type virus particle formation in these placentas are unresponsiveness of the trophoblast to interferon or inadequate exposure time to interferon of placental sites from which C-type particles are produced. PMID- 2559169 TI - The effect of angiotensin converting enzyme inhibitors on isolated glomeruli. AB - The following study was designed to examine the effect of the angiotensin converting enzyme (ACE) inhibitors captopril and enalapril on certain biological and physiological activities of isolated preparations of glomeruli from normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive SHR rats. The results suggest that the contractile response of glomeruli from SHR to exogenous angiotensin II (Ang II) is significantly enhanced by captopril whereas it has a different effect on those from normotensive (WKY) rats. These effects are unrelated to changes in glomerular Ang II binding or the level of plasma renin activity (PRA), but they do depend partially on the sodium status of the animal. Whereas captopril induced a modest increase in the basal rate of prostaglandin E2 (PGE2) synthesis by glomeruli from normotensive (WKY) but not by those from SHR, the reverse occurred in the presence of Ang II. Prostaglandin E2 synthesis was increased in both strains of rats in the presence of excess arachidonic acid. Further increases occurred with the addition of captopril, these increases being significantly greater by glomeruli from WKY rats than by those from SHR, while enalaprilat had no effect on PGE2 production. The glomerular synthesis of PGE2 was not influenced in either strain of rats by in vitro administration of captopril. PMID- 2559170 TI - Angiotensin converting enzyme inhibition and renin release from the kidney. AB - Experiments were performed in anaesthetized cats, to determine whether renal nerves interfere with the negative feedback action of angiotensin II (Ang II) on renin release. The increase in renin release from the innervated kidney in response to captopril-induced inhibition of Ang II generation was compared with the response of the contralateral denervated kidney. Renin release was measured before (control), and 5, 15 and 30 min after the beginning of captopril infusion (1 mg/kg priming dose followed by 1 mg/kg per h for 30 min intravenously), and 60 min after the end of the infusion. During the captopril treatment renin release from both kidneys increased, but after 15 and 30 min the increase in renin release from the innervated kidneys was significantly larger than that observed in the denervated kidneys. After the captopril infusion was stopped, renin release from both kidneys returned towards control values. These results could not be explained on the basis of the changes in renal haemodynamics, excretory functions and efferent renal nerve activity observed during the captopril infusion. The data suggest that the renal nerves influence the changes in renin release caused by captopril by increasing the sensitivity of juxtaglomerular cells to the negative feedback action of Ang II. PMID- 2559171 TI - Molecular biology and pathophysiology of the intrarenal renin-angiotensin system. AB - There has been considerable interest in the existence of an intrarenal renin angiotensin system and its physiological implications. Recent demonstrations of renin, angiotensinogen and angiotensin converting enzyme messenger (m)RNAs in the kidney have provided strong evidence for the presence of an independent local system. This has been further supported by the demonstration of tissue-specific regulation of renin and angiotensinogen mRNA expression which may lead to differential systemic and intrarenal angiotensin activities. Using in situ hybridization, we have localized the intrarenal sites of gene expression and possible angiotensin production. One major site appears to be the proximal tubule, where local angiotensin can regulate sodium reabsorption and urine pH. Renin and angiotensinogen mRNA expressions are regulated by several common factors. In particular, sodium depletion stimulates the expression of both genes in the kidney, increasing the production of intrarenal angiotensin that is important in maintaining sodium homeostasis. Renal renin and angiotensinogen mRNA levels are altered in experimental heart failure and the spontaneously hypertensive rat (SHR). These changes in intrarenal renin and angiotensinogen mRNA expression may be important in the renal pathophysiology of these diseases. PMID- 2559172 TI - Restraining effect of captopril on cardiovascular sympathetic efferent neural activity. AB - In 13 decerebrate cats we studied the effects of captopril (10 mg/kg, intravenous bolus) on the background cardiac preganglionic sympathetic discharge and on its responsiveness to brief baroreceptor deactivations induced by premature ventricular contractions. Captopril caused a significant reduction of 57 +/- 7% (from 2.4 +/- 0.4 impulses/0.1s) in sympathetic discharge, with a mean latency of 35 +/- 3 min from the time of drug administration. In addition, the increase in cardiac sympathetic firing during premature ventricular contractions was markedly reduced, from 257 +/- 30% to 70 +/- 17%, before and after administration of captopril. These data indicate that captopril is not only likely to inhibit sympathetic discharge to the heart, but is also likely to restrain the excitatory sympathetic responses to baroreceptor deactivation. PMID- 2559173 TI - Angiotensin converting enzyme inhibition delays the progression of chronic renal failure in hypertensive patients with immunoglobulin A nephropathy. AB - Ten hypertensive, non-nephrotic patients with immunoglobulin (IgA) nephropathy treated with conventional therapy (vasodilators, beta-blockers, diuretics) were observed for 21.4 +/- 9 months. Treatment was then changed to an angiotensin converting enzyme (ACE) inhibitor alone for an additional 22.6 +/- 9 months. The ACE inhibitor therapy was associated with a lower diastolic blood pressure and a slowing in the rate of decline in renal function, but no change in proteinuria. PMID- 2559174 TI - The angiotensin converting enzyme in the kidney. AB - Immunohistochemical studies and experiments with microdissected nephron segments indicate that the angiotensin I converting enzyme (ACE) in the kidney is expressed in the vascular endothelial cells of the renal vessels and in the epithelial cells of the proximal convoluted tubule and the pars recta. Angiotensin converting enzyme is a membrane-bound zinc metallopeptidase and the primary structure has recently been determined by protein sequencing and molecular cloning. It is probably anchored to the cell membrane by a single, short, transmembrane domain located near the carboxy-terminal extremity. The larger, externally situated, amino-terminal part of the molecule is organized in two large, highly homologous domains, each with a putative active site. The function of the endothelial enzyme in the renal vessels is primarily related to angiotensin II (Ang II) formation. However, its level of expression in renal vessels, especially at the glomerular level, appears to be very low in the adult human kidney, and there is evidence that the conversion of angiotensin I (Ang I) may be a rate-limiting step in Ang II formation in the kidney. The vascular enzyme may also contribute to the inactivation of kinins in the peritubular circulation. In the epithelial cells of the proximal tubule, ACE is present in both the brush border and the basolateral membrane. Although the basolateral enzyme may be involved in Ang II formation in the peritubular interstitium, the function of the enzyme on the brush border is unknown. The effects of ACE inhibitors on renal function are primarily, if not exclusively, related to Ang II suppression and perhaps kinin potentiation in the renal circulation. PMID- 2559176 TI - [Expression of an Epstein-Barr virus early antigen and cell fusion]. AB - Superinfection of Raji cells with Epstein-Barr virus (EBV) derived from P3HR-1 (P3H-EBV) leads to syncytium formation. Studies using inhibitors of DNA and protein synthesis suggest that the fusion-inducing factor belongs to the early antigen of virus-specified proteins. Induction of early antigen (EA) in Raji cells by using various chemicals also leads to syncytium formation. However, A2L/AH hybrid cells, which were prepared by fusion between epithelial cells of adenoid origin and lymphocytes transformed by B95-8 virus, could not be superinfected with P3H-EBV. But, the syncytium formation was observed when EBV-EA was induced by chemicals in A2L/AH cells. It was considered that the synthesis of viral protein in superinfected Raji cells is essential for fusion and that the viral genes, inducible in Raji and A2L/AH cells by treatment with chemicals code for the necessary functions that lead to syncytia formation. PMID- 2559175 TI - Cytomegalovirus myocarditis in a heart transplant recipient: sensitive monitoring of viral DNA by the polymerase chain reaction. AB - A 54-year-old female heart transplant recipient had an acute episode of graft rejection that was treated with high-dose immunosuppression therapy. During this therapy a second febrile illness developed, which was accompanied by hypotension, anemia, and rash. Findings for subsequent myocardial biopsy specimens were negative for cytomegalovirus by culture but were strongly positive for cytomegalovirus deoxyribonucleic acid by the polymerase chain reaction. Histologic observation of viral inclusions in the myocardial fibers supported the diagnosis of cytomegalovirus myocarditis. The polymerase chain reaction therefore can provide a rapid and highly sensitive method for heart transplant patients with suspected cytomegalovirus myocarditis. PMID- 2559179 TI - Histopathological grading of cervical intraepithelial neoplasia (CIN)--is there a need for change? PMID- 2559177 TI - Properties of benzodiazepine binding sites in peripheral blood lymphocytes. AB - Benzodiazepine (BDZ) binding sites were studied by using 3H-diazepam and 3H-Ro 5 4864 in intact lymphocytes from peripheral blood (PBL), in comparison to kidney and cerebellum. Experiments with 3H-diazepam performed at equilibrium and measuring kinetics revealed that BDZ binding sites are indeed present in rat PBL. The binding is saturable (Bmax 557 fmoles/10(6) cells), with high affinity (KD = 9.3 nM) and reversible. Specific binding sites are also observed by saturation experiments with 3H-Ro 5-4864 (Bmax 175 fmoles/10(6) cells, KD 2.2 nM). In addition, analysis of saturation isotherms obtained with 3H-diazepam indicates that BDZ binding sites are also present in human PBL. Scatchard plot of binding isotherms revealed an apparent single population of sites in all cases. The pharmacological characterization of BDZ binding sites in PBL, as compared with those of kidney and cerebellum, showed that these sites belong to the so-called "peripheral type." PMID- 2559178 TI - Detection of cytomegalovirus-specific IGM in renal transplant recipients. AB - We have compared two IgM-specific cytomegalovirus (CMV) antibody assays, an immunofluorescence assay (IFA-M) and an enzyme-linked antigen immunoassay (ELA M), with an assay for CMV total antibody (ELISA) and viral culture for the detection of active CMV infection in renal transplant recipients. Of 75 patients (49 ELISA negative pretransplant, 26 ELISA positive), CMV-specific IgM was detected in 35 (27 ELISA negative pretransplant, 8 ELISA positive) using the IFA M assay and in 25 (16 ELISA negative pretransplant, 9 ELISA positive) using the ELA-M test. Of the 25 patients identified as positive by ELA-M, 21 had positive viral cultures post-transplant, two seronegative patients had evidence of infection indicated by post-transplant seroconversion, and two patients were seropositive pretransplant but remained viral culture negative throughout the follow-up period. ELA-M and CMV total antibody ELISA detected primary infection in renal transplant recipients equally well, but ELA-M was found to be superior to ELISA and IFA-M for detecting reinfection and reactivation infections. PMID- 2559180 TI - A simple micropore system for experimental studies on trichomonad parasites. AB - A simple leak-free micropore chamber containing protozoan parasite species was implanted subcutaneously on the back of hamsters and evaluated for viability and multiplication of protozoan parasites. Trophozoites of defined strains of Entamoeba histolytica, Giardia lamblia, Trichomonas vaginalis, and Tritrichomonas foetus were used; their survival and multiplication in the chambers formed the basis of evaluation. Entamoeba histolytica and G. lamblia did not survive more than 6 hr and succumbed due to cellular adhesion. Trichomonas vaginalis and T. foetus survived 3 and 6 days and multiplied a maximum of 3.6 and 26 times, respectively. This indicated that exchange of body fluids and cells needed for the survival and multiplication of trichomonads readily occurs. This preliminary observation showed that micropore chambers may be useful for chemotherapeutic and immunological studies on trichomonads in ectopic sites. PMID- 2559182 TI - Partial purification of gelatinases and effect of anti-inflammatory drugs on the tumor necrosis factor-stimulated production of active gelatinase by granulation tissue in culture. AB - Gelatinases produced by granulation tissue in culture were separated into two fractions when the conditioned medium was chromatographed on diethylaminoethyl Sephacel; one contained latent and active gelatinases with molecular weight of about 74 kDa (low-Mr gelatinase), and the other contained an active gelatinase with molecular weight of about 112 kDa (high-Mr gelatinase) as estimated by gel filtration on Sephadex G-150. The former was unbound and the latter was bound to Zn-chelating Sepharose. Both low- and high-Mr gelatinases, however, occurred in mainly two bands corresponding to molecular weights of 64 and 57 kDa on sodium dodecyl sulfate-substrate polyacrylamide gel electrophoresis. Recombinant human tumor necrosis factor-alpha (TNF) markedly enhanced the production of gelatinases, especially high-Mr active gelatinase. Dexamethasone and hydrocortisone strongly suppressed TNF-mediated production of high-Mr active gelatinase, whereas indomethacin, piroxicam and mepacrine had no effect. The results suggest that steroidal anti-inflammatory drugs suppress a rapid collagen breakdown in granulation tissue through their inhibitory actions--one of which is the suppression of active gelatinase production enhanced by cytokines including TNF. PMID- 2559183 TI - Hydrolysis of 3',5'-dioctanoyl-5-fluoro-2'-deoxyuridine (FdUrd-C8) in rabbit and human plasma and its release from oily solution. AB - Hydrolysis of 3',5'-dioctanoyl-5-fluoro-2'-deoxyuridine (FdUrd-C8), a lipophilic prodrug of 5-fluoro-2'-deoxyuridine (FdUrd), 3'-octanoyl-5-fluoro-2' deoxyuridine(3'-octanoyl FdUrd) and 5'-octanoyl-5-fluoro-2'-deoxyuridine (5' octanoyl FdUrd) in the rabbit and human plasma and release of FdUrd-C8 and its hydrolyzed species from Lipiodol Ultra-Fluid (Lipiodol), an oily lymphographic agent, containing FdUrd-C8 were examined. The rates of hydrolysis of FdUrd-C8, 3' octanoyl FdUrd and 5'-octanoyl FdUrd in the rabbit plasma were fast and almost the same among the three compounds; half lives were 2.4, 3.6 and 3.9 min for 5' octanoyl FdUrd, FdUrd-C8 and 3'-octanoyl FdUrd, respectively. In the human plasma, however, the rates of hydrolysis were much different among the three compounds and were slower than those in the rabbit plasma; half lives were 11.5, 130 and 1020 min for 5'-octanoyl FdUrd, FdUrd-C8 and 3'-octanoyl FdUrd, respectively. Ratios of the rate constant of 5'-octanoyl FdUrd to that of 3' octanoyl FdUrd were 1.6 and 85.7 in the rabbit plasma and in the human plasma, respectively. In a release study, although detected species in a release medium were different between the rabbit plasma and the human plasma and the amount of each species reflected the characteristics of esterase in each plasma, the total amounts of compounds released were almost the same both in the rabbit plasma and the human plasma.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559181 TI - Comparison of decalcified freeze-dried bone allograft and porous particulate hydroxyapatite in human periodontal osseous defects. AB - The purpose of this study was to clinically compare the healing potential of the osteoinductive decalcified freeze-dried bone allograft (DFDBA) with an osteoconductive synthetic graft, particulate porous hydroxyapatite (HA). Six patients ranging in age from 28 to 52 (mean age 42.6) participated in this investigation. They were without significant systemic disorders and had advanced periodontitis with at least two comparable periodontal defects. Each patient received initial therapy consisting of oral hygiene instruction, scaling, root planing, and occlusal adjustment as indicated. Probing pocket depth, gingival recession, attachment levels, and bleeding on probing were recorded in the posthygiene phase of therapy. Alveolar crest height and depth of osseous defect were obtained at the time of surgery. The cemento-enamel junction was used as the fixed reference point. All measurements were repeated at the time of a 6-month reentry. There was no significant difference in any of the soft tissue measurements when DFDBA and HA were compared. However, both treatment modalities reduced pocket depth and demonstrated a gain in clinical attachment levels. There was 2.2 mm of bone repair with DFDBA and 2.1 mm with HA. These values corresponded to a percent defect fill of 61% for DFDBA and 53% for HA. These values were likewise not statistically different. PMID- 2559184 TI - Three binding sites of 125I-iodocyanopindolol, i.e. beta 1, beta 2-adrenergic and 5HT1B-serotonergic receptors in rat brain determined by the displacement and Scatchard analysis. AB - The selectivity of binding of 125I-iodocyanopindolol (125I-ICYP) to beta 1-, beta 2-adrenergic and 5HT1B-serotonergic receptors were analysed using both the Scatchard and displacement methods. When the binding of 125I-ICYP to the rat brain membrane was examined by means of the displacement analysis with l metoprolol, triphasic displacement curves were observed. Analyses carried out on the basis of the displacement of 125I-ICYP by beta-selective antagonists, alpha adrenergic agonists and antagonists, and serotonergic agonists and antagonists indicated the correspondence of the super high, high- and low-affinity sites of 125I-ICYP binding to beta 2-, beta 1-adrenergic and 5HT1B-receptors, respectively. By contrast, the Scatchard analysis revealed the biphasic character of the 125I-ICYP binding and the complete inhibition of the binding to high- and low-affinity sites by l-metoprolol (30 microM) and 5-hydroxytryptamine (5HT, 10 microM), respectively, suggesting the correspondence of the high- and low affinity sites to beta-adrenergic and 5HT1B-receptors. Thus, regarding the determination of the selectivity of binding of a radioligand, the displacement analysis is more informative than the Scatchard analysis. In the case of 125I ICYP the separation of beta 1- and beta 2-adrenergic receptors becomes feasible only with the displacement method. PMID- 2559185 TI - Inter-organ relation between salivary gland and kidney in lithium excretion. II. Salivary, renal and systemic clearances of lithium under continuous stimulation of salivation in water loaded dogs. AB - This study was undertaken to investigate the effects of water loading on lithium clearance in dogs under the continuous stimulation of salivation as well as to clarify the mechanism of the inter-organ relation between salivary gland and kidney in lithium excretion. Dogs were given intravenously 0.145 meq/kg of lithium chloride followed by the continuous stimulation of salivation with citric acid solution. Fifty ml of water was loaded orally 7 times at 1-h intervals. Parotid and mandibular-sublingual salivas were collected separately by means of permanent fistulae. (1) Plasma concentrations of lithium were not significantly different from either those in the experiment with continuous stimulation under no water loading or those in the control experiment without continuous stimulation. (2) Salivary clearance of lithium was markedly increased compared with that in the control experiment. The urinary flow rate, which was decreased under the continuous stimulation of salivation without water loading, was restored by the water loading. The renal clearance of lithium, which was also decreased under the continuous stimulation, remained at the reduced level under the condition of this study. Consequently, the systemic clearance of lithium did not change from that in the experiment under the continuous stimulation without water loading or that in the control experiment. (3) It was suggested that the salivary and renal excretion mechanisms of lithium might be similar to those of potassium, since a similar interrelation between salivary and renal clearances was observed for potassium. (4) It was suggested that the reduction in the renal clearance of lithium under the continuous stimulation of salivation was attributed to the sodium loss caused by the excessive salivation. PMID- 2559186 TI - Selectivity for opioid receptor subtypes of enkephalin analogues in isolated smooth muscle and in the analgesic effect in mice. AB - Selectivity for opioid receptor subtypes of enkephalin analogues (KK-1, -2, -3 and -4) of Tyr moiety on the N-terminal, and Phe-ol group on the C-terminal, connected with the methylene group (n = 1-4) were examined in isolated smooth muscle preparations and in the analgesic effect in mice. In the longitudinal muscle preparations of guinea pig ileum (GPI), morphine, U-50488H and all the enkephalin analogues inhibited electrically evoked contractions, and the inhibitory effects of morphine, KK-1, KK-2 and KK-3 were antagonized by naloxone with relatively high pA2 values, while that of U-50488H and KK-3 were preferentially antagonized by norbinaltorphimine. In the rabbit vas deferens preparations (RVD), on the other hand, U-50488H, KK-3 and KK-4 showed weak inhibitory effects and the inhibition of U-50488H and KK-3 were antagonized by norbinaltorphimine. By intracerebroventricularly (i.c.v.) injection, enkephalin analogues produced analgesia in the acetic acid (AcOH) writhing test, and the effect of KK-1 and KK-2 as well as morphine was antagonized by 1 mg/kg naloxone, while those of U-50488H and KK-3 were sensitive to 1 mg/kg Mr2266. In conclusion, enkephalin analogues with a short methylene chain between the functional groups, KK-1 and KK-2, mainly exert their effect through opioid mu-receptors, while those of longer chain, KK-3 and KK-4, act through kappa-receptors preferentially, and KK-3 is situated in the alternating point of the selectivity for mu- and kappa receptors. PMID- 2559187 TI - [Genetic and biochemical studies on the mechanism of mammalian chromosome replication]. AB - It has been clarified that basic mechanism of deoxyribonucleic acid (DNA) replication is conserved from bacteria to higher cells. What distinguishes prokaryotic and eukaryotic modes of DNA replication most clearly is that bacterial chromosomes form a single replicon copied from a single initiation point and eukaryotic chromosomes consist of multiple replicons that initiate at multiple points. Thus, eukaryotes have to coordinate orderly replication of the genome. In order to understand this complex problem as a whole, three approaches were chosen. First approach is a genetic one. Certain number of temperature sensitive (ts) mutants were isolated from mouse FM3A cells. One of the ts mutants, designated as tsFT20, was shown to contain heat-labile DNA polymerase alpha (pol. alpha). By the use of this mutant strain, it was proved that pol. alpha is essential for mammalian DNA replication. In addition, the human gene for pol. alpha on the X chromosome was assigned. Second approach is an enzymological one. FM3A cells were used for the identification and characterization of enzymes and proteins supposed to be involved in DNA replication. Four DNA-dependent ATPases, three pol. alpha stimulation factors, DNA topoisomerases I and II have been identified, as well as a stimulation factor for the assembly of nucleosome. DNA helicase activity was detected in two of the DNA-dependent ATPase (B and C1). Third approach is the reconstitution of DNA replication in cell-free system. By use of polyoma virus DNA as a template, cell-free extract from FM3A cells supported DNA replication in the presence of polyoma virus large T-antigen. This cell-free system will be useful for the analysis of the function of replication enzymes and proteins as well as the characterization of ts mutants. PMID- 2559189 TI - [Fractional determination of bile acids bound with protein in serum by high performance liquid chromatography using dual-column switching system]. AB - A simple and rapid technique for the fractional determination of bile acids bound with protein in the serum was developed by using high performance liquid chromatography with a dual-column switching system. The serum samples were directly injected onto a first column (hydroxyapatite), which was initially flushed with 1 mM phosphate buffer. Serum proteins were strongly retained on a column of hydroxyapatite, but free bile acids were not retained. The bile acids were adsorbed on a second column (Serumout-25) and eluted onto a column of immobilized 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD) with an elution solvent (CH3CN-MeOH-30 mM ammonium acetate, 30:30:40, v/v/v). Reduced nicotinamideadenine dinucleotide was produced on the immobilized 3 alpha-HSD column and then determined fluorometrically. Subsequently, the hydroxyapatite column was flushed with 20 mM phosphate buffer. Bile acids bound with albumin were eluted and condensed on Serumout-25. The phosphate buffer (400 mM) was finally used for the elution of bile acids bound with globulin from the hydroxyapatite column. Each condensed bile acid was eluted onto the immobilized 3 alpha-HSD column as described above. PMID- 2559188 TI - [Anti-tumor promoting activities and inhibitory effects on Epstein-Barr virus activation of Shi-un-kou and its constituents]. AB - The Kampo-prescription, Shi-un-kou, and its constituent crude drugs [Lithospermum erythrorhizon (1), Macrotomia euchroma (2) and Angelica acutiloba (3)] were assayed for their inhibitory effects on Epstein-Barr virus activation induced by the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA). The crude drugs exhibited inhibitory activity singly and in combinations. In particular, the combination of 2 and 3 yielded enhanced inhibition and lower cytotoxicity. The anti-tumor promoter activity suggested by these results was further investigated in an in vivo study, which demonstrated that Shi-un-kou markedly inhibited TPA induced skin tumor formation in mice. PMID- 2559190 TI - Pharmacokinetics of an ACE inhibitor, S-9780, in man: evidence of tissue binding. AB - Pharmacokinetic data from 20-min constant rate infusions of the ACE inhibitor S 9780 1 mg to 16 subjects were studied for evidence of nonlinearity. A hierarchy of standard compartmental models and of nonlinear binding models was fitted to the data by least squares nonlinear regression and the most appropriate model was chosen on the basis of F-ratio tests, Schwarz criteria, and residual plots. A one compartment model which included saturable tissue and plasma binding components allowed the best overall description of the data. Median parameter estimates from this model suggest that approximately 308 nmol of plasma binding sites and 572 nmol of tissue binding sites were present and that the total plasma concentration of S-9780 at 50% saturation of binding sites was 16.5 nmol L-1. The elimination half-life for free drug in plasma was only 30 min. This model describes the discrepancy previously noted between accumulation and apparent elimination half lives for long-acting ACE inhibitors and offers a noninvasive method for assessment of tissue-bound ACE inhibitor in vivo. PMID- 2559191 TI - Intestinal absorption mechanism of dipeptide angiotensin converting enzyme inhibitors of the lysyl-proline type: lisinopril and SQ 29,852. AB - The intestinal absorption mechanism of two nonsulfhydril lysyl-proline angiotensin converting enzyme (ACE) inhibitors, lisinopril (1) and SQ 29,852 (2; [(S)-1-[6-amino-2-[[hydroxy (4-phenylbutyl)-phosphinyl]oxy[-1-oxohexyl]-L proline) were investigated in rats using a single-pass perfusion method. Compound 2 is well absorbed from rat jejunum, whereas lisinopril absorption is relatively low. The permeability of both ACE inhibitors is concentration dependent and is decreased by the dipeptide Tyr-Gly and by cephradine, indicating a nonpassive absorption mechanism via the peptide carrier-mediated transport system. Compound 2 is well absorbed by a nonpassive mechanism, in parallel with a small passive component. The estimated dimensionless carrier parameters for 2 are J*max = 0.16, Km = 0.08 mM, P*c = 2.0, and P*m = 0.25; for lisinopril, passive absorption is not significant and its absorption is nonpassive: J*max = 0.032, Km = 0.082 mM, and P*c = 0.39 (where J*max is the maximal flux, Km is the Michaelis constant, P*c is the carrier permeability, and P*m is the passive permeability). These results offer a mechanistic explanation for the prolonged ACE inhibition and the low oral bioavailability of lisinopril, and for the nonlinear pharmacokinetics of 2. PMID- 2559192 TI - Voltage-dependent potassium channels in mouse Schwann cells. AB - 1. Ionic currents in Schwann cells cultured from enzymatically dissociated sciatic nerves of newborn mice were recorded by the whole-cell variation of the patch-clamp technique. 2. In these cells only the voltage-dependent K+ currents were recorded. The K+ current was suppressed by quinine, 4-aminopyridine (4-AP) or tetraethylammonium (TEA), their half-suppression concentrations being 22 microM, 0.3 mM and 15 mM, respectively. 3. The peak amplitudes and density of the K+ currents in these Schwann cells increased rapidly during the first 2 days of the culture. 4. In an investigation of the linkage between K+ channels and Schwann cell proliferation, three different K+ channel blockers (quinine, 4-AP and TEA) were added to the medium at different stages of the culture. In media containing sublethal doses of quinine or 4-AP, the start of cell proliferation was delayed when these drugs were added at 12 h or on day 3. The same doses of these drugs applied on day 6, when the Schwann cells were proliferating, did not affect cell proliferation. TEA showed a discrepancy between the dose-dependent blocking of K+ channels and cell proliferation because of its additional cytotoxic effects. 5. It is concluded that voltage-dependent K+ channels in mouse Schwann cells are similar to those observed in human and murine T lymphocytes. These K+ channels are suggested to be involved in Schwann cell proliferation at early stages of development. PMID- 2559193 TI - An inwardly directed electrogenic sodium-bicarbonate co-transport in leech glial cells. AB - 1. We have used double-barrelled ion-sensitive microelectrodes to measure the intracellular pH, pHi, the intracellular Na+ activity, aiNa, and the membrane potential in identified glial cells of the central nervous system of the leech Hirudo medicinalis to study the effect of CO2-HCO3-. 2. When a HEPES-buffered saline was exchanged for a saline buffered with 2% CO2 + 11 mM-HCO3-, keeping the pH constant at 7.4, the mean steady-state pHi of the glial cells increased from 6.85 +/- 0.06 to 7.18 +/- 0.13 (mean +/- S.D., n = 25). 3. This CO2-HCO3- dependent alkalinization was inhibited in the absence of external Na+ (exchanged by N-methyl-D-glucamine), but was unaffected by the inhibitor of Na+-H+ exchange, amiloride (2 mM). 4. The aiNa of the glial cells increased by 2-4 mM from a mean steady state of 7.2 +/- 2 mM (mean +/- S.D., n = 6) upon introduction of CO2-HCO3 -buffered saline. This CO2-HCO3- -dependent rise in aiNa increased to about double when the pHi had been decreased by acid loading the cells (addition and subsequent removal of NH4+). 5. The CO2-HCO3- -dependent increases of pHi and aiNa were inhibited by the stilbene 4,4-diisothiocyanostilbene-2,2'-disulphonic acid (DIDS, 0.5-1.0 mM). 6. Removal of external Cl- and depletion of intracellular Cl- did not inhibit the CO2-HCO3- -dependent alkalinization. 7. The CO2-HCO3- -dependent alkalinization was unaffected by inhibitors of the carbonic anhydrase, acetazolamide (0.2 mM) or ethoxzolamide (2 microM). 8. The membrane potential became more negative by 3-20 mV upon addition of CO2-HCO3-. This hyperpolarization was even further enlarged in the presence of Ba2+ (which reduces the K+ permeability) or at increased external K+ concentration (which depolarizes the membrane and brings the membrane potential to the K+ equilibrium potential). The CO2-HCO3- -induced membrane hyperpolarization was inhibited in Na+-free saline and in the presence of DIDS. Ouabain (0.5 mM) sometimes reduced, but never abolished, the hyperpolarization. 9. The stoichiometry of the co transport is suggested to be 2 HCO3-:1 Na+ with an equilibrium potential of -90 mV calculated for this coupling ratio in the steady state. 10. It is concluded that in the presence of CO2-HCO3- an inwardly directed electrogenic Na+-HCO3- co transport is stimulated across the glial membrane, which greatly determines the pHi and thereby affects the intracellular buffering power of the glial cells. PMID- 2559194 TI - Comparison between sympathetic adrenergic and purinergic transmission in the dog mesenteric artery. AB - 1. Electrical transmural stimulation evoked a sympathetic contraction in the isolated dog mesenteric artery. This contraction consisted of adrenergic and purinergic components, which were separately observed under conditions where postjunctional P2 purinoceptors were desensitized with alpha, beta-methylene ATP or postjunctional alpha 1 adrenoceptors were blocked by prazosin, respectively. 2. The purinergic component was transient and developed immediately after the start of stimulation and declined rapidly after a peak. In contrast, the adrenergic contraction slowly developed and lasted longer than the purinergic component. Thus, the purinergic and adrenergic components predominantly contributed to the early and later phases of the total sympathetic response, respectively. 3. The peak amplitudes of contraction of both components were similar at 1 Hz, but the adrenergic component occurred more dominantly in the responses to higher frequency stimulation. 4. Cocaine, a neuronal uptake inhibitor of noradrenaline, potentiated the adrenergic component but attenuated the purinergic component. This attenuation was reversed by DG-5128, a prejunctional alpha 2 adrenoceptor antagonist. Treatment with DG-5128 alone augmented both adrenergic and purinergic components. 5. 8-Phenyltheophylline, a P1 purinoceptor antagonist, potentiated the purinergic component without affecting the adrenergic component. 6. Exogenous noradrenaline produced a sustained contraction, which was potentiated by cocaine and was competitively inhibited by prazosin. alpha, beta-Methylene ATP and 8-phenyltheophylline had no effect on the response to noradrenaline. Exogenous ATP produced a transient contraction, which was abolished under conditions where postjunctional P2 purinoceptors were desensitized with alpha, beta-methylene ATP. 8 Phenyltheophylline potentiated but cocaine or prazosin did not affect the ATP response. 7. Electrical stimulation produced an increase in 3H efflux from the sympathetic nerve terminals in the mesenteric arteries pre-incubated with [3H]noradrenaline. The evoked efflux was significantly augmented by cocaine or DG 5128 and was inhibited by guanethidine or tetrodotoxin. alpha, beta-Methylene ATP and 8-phenyltheophylline were without effect. Adenosine reduced the 3H efflux and the inhibition was suppressed by 8-phenyltheophylline. 8. These results suggest that sympathetic contraction of the dog mesenteric artery is caused through not only adrenergic but also purinergic mechanisms, and that both the sympathetic transmissions are predominantly modulated through prejunctional adrenergic mechanisms. PMID- 2559195 TI - Possible involvement of prostaglandin E in development of ACTH response in rats induced by human recombinant interleukin-1. AB - 1. Intravenous (I.V.) injection of human recombinant interleukin-1 alpha (IL-1 alpha) produced dose-dependent monophasic fevers in rats. Moreover, the I.V. injection of IL-1 alpha produced dose-dependent rises in the plasma concentrations of adrenocorticotrophic hormone (ACTH) 30 min after injections with dosages of 5 micrograms/kg and 15 micrograms/kg of IL-1 alpha. 2. The febrile responses induced by the I.V. injection of IL-1 alpha (15 micrograms/kg) were completely abolished, and conversely hypothermia occurred, when the animals were pre-treated with a cyclo-oxygenase inhibitor, indomethacin (INDO). Pre treatment with INDO also inhibited the increase in the plasma concentrations of ACTH induced by I.V. injection of IL-1 alpha (15 micrograms/kg), indicating that enhancement of plasma concentrations of ACTH induced by I.V. injection of IL-1 alpha is processed through the action of prostaglandins. 3. Intrapreoptic injection of prostaglandin E2 produced a dose-dependent fever with a rapid onset at doses of 25 and 100 ng. Moreover, the intrapreoptic injection of prostaglandin E2 increased the plasma concentrations of ACTH in a dose-dependent manner 30 min after injections. 4. The intrapreoptic injection of IL-1 alpha (20 ng) caused slow monophasic fever. However, no significant elevation of plasma concentrations of ACTH was observed 30, 90 and 180 min after the intrapreoptic injection of IL-1 alpha, as compared with the ACTH levels at each time in the control group which received an intrapreoptic injection of saline. 5. These results suggest that intrapreoptic prostaglandin E plays an important role in the ACTH response by inducing the release of corticotrophin-releasing factor (CRF). PMID- 2559196 TI - Central neurotensin nerves modulate colo-colonic reflex activity in the guinea pig inferior mesenteric ganglion. AB - 1. The effects of neurotensin and of stimulation of preganglionic nerves on peripheral afferent synaptic input from segments of distal colon to neurones in the inferior mesenteric ganglia of guinea-pigs were studied using intracellular recording techniques in vitro. 2. Electrical stimulation of colonic afferent nerve fibres evoked fast, nicotinic synaptic responses (fast EPSPs or action potentials) followed by a slow depolarizing response (slow EPSP). 3. Neurotensin (1 microM) increased the amplitude and duration of slow EPSPs evoked by stimulation of colonic afferents. 4. Distention of a segment of distal colon left attached to an inferior mesenteric ganglion evoked a slow depolarization. Neurotensin (1 microM) increased the amplitude and duration of distention-induced depolarizations. 5. Electrical stimulation of central preganglionic nerve fibres present in the third and fourth lumbar ventral roots increased the amplitude and duration of slow EPSPs evoked by electrical stimulation of colonic afferent nerves. This facilitatory effect was abolished after desensitization to neurotensin. 6. Slow depolarizations evoked by neurotensin and by stimulation of central preganglionic nerves converted subthreshold fast EPSPs due to mechanosensory synaptic input from an attached segment of distal colon to action potentials. This increase in firing rate of sympathetic ganglion cells led to a decrease in colonic intraluminal pressure. 7. Taken together these data support the hypothesis that neurotensin or a closely related substance contained in central preganglionic nerves facilitated release of a non-cholinergic excitatory transmitter from colonic mechanosensory nerves. The slow depolarization evoked by the non-cholinergic transmitter converted on-going subthreshold fast EPSPs to action potentials thereby increasing sympathetic output to the colon. 8. It is suggested that under normal in vivo conditions, central preganglionic fibres containing neurotensin or a closely related peptide modulate peripheral reflex activity through prevertebral ganglia in guinea-pigs. PMID- 2559197 TI - Endothelium-derived relaxing factor inhibits the formation of inositol trisphosphate by rabbit aorta. AB - 1. The effects of endothelium-derived relaxing factor (EDRF), sodium nitroprusside, 8-bromo-cyclic GMP and atrial natriuretic factor (ANF) on inositol trisphosphate (IP3) levels were studied in isolated rabbit aortic preparations stimulated with noradrenaline. 2. In endothelium-containing preparations, acetylcholine, which stimulated EDRF release, inhibited noradrenaline-stimulated IP3 formation. The EDRF inhibitor haemoglobin reversed this effect. 3. In endothelium-denuded preparations, sodium nitroprusside, 8-bromo-cyclic GMP and ANF each similarly inhibited the rise in IP3 levels stimulated by noradrenaline. 4. These findings show that in rabbit aorta, agents which increase cyclic GMP inhibit the noradrenaline-induced rise in IP3 levels and may provide an explanation for the previously reported observations that cyclic GMP inhibits the noradrenaline-stimulated increase in calcium influx and release of intracellular calcium in vascular smooth muscle. PMID- 2559199 TI - Tetrodotoxin block of single germitrine-activated sodium channels in cultured rat cardiac cells. AB - 1. The open time of single Na+ channels in excised (outside-out) patches from cultured late-fetal rat ventricular myocytes was prolonged to several minutes by germitrine (0.5 mM) in order to analyse tetrodotoxin (TTX) blocking kinetics. 2. The germitrine modification appeared during depolarizing pulses that activated normal Na+ channels. Following repolarization to -100 mV, the modified Na+ channel remained activated for 136 +/- 186 s (mean +/- S.D., n = 54) with an open channel current amplitude of -0.5 pA. The predominant open state with a mean open time of 0.13 s was interrupted by brief closing events lasting for milliseconds. Replacing extracellular Na+ by Cs+ decreased the current amplitude to -0.1 pA. 3. Extracellular superfusion with TTX (3 x 10(-7) M) of a single germitrine activated Na+ channel induced full channel closures lasting seconds (blocked events) separated by channel reopenings (unblocked events) that were indistinguishable in terms of amplitude and gating kinetics from the germitrine activated state in the absence of TTX. 4. Cumulative probability histograms of blocked and unblocked events (n greater than 140) collected during long-lasting germitrine modifications at 10(-7) and 3 x 10(-7) M-TTX are well described by single exponentials. The 3-fold increase in [TTX] decreased the time constant of the unblocked state, tau o, from 11.9 to 4.7 s, while the time constant of the blocked state, tau c, was not significantly altered from 8.6 to 9.7 s. A microscopic association rate constant of 7.7 x 10(5) M-1 s-1, dissociation rate constant of 0.11 s-1, and equilibrium dissociation constant of 1.4 x 10(-7) M (at -100 mV) were calculated (20 degrees C). 5. Increasing [TTX] to 10(-5) M decreased tau o to 86 ms. This argues against the existence of a slower conformational step interposed between the binding of TTX to an open channel and the resultant channel closure. 6. Setting the membrane potential to -50 or 0 mV subsequent to a germitrine modification at -100 mV did not significantly alter TTX (3 x 10(-7) M) blocking kinetics: tau o was 6.7 s at -50 mV and 5.2 s at 0 mV; tau c was 8.9 and 8.1 s, respectively. 7. These results suggest that blocked events correspond to the random times that a TTX molecule resides on the Na+ channel before it dissociates, and unblocked events correspond to the random waiting times of an unoccupied channel before it binds another toxin molecule.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2559198 TI - Effect of nocodazole on the water permeability response to vasopressin in rabbit collecting tubules perfused in vitro. AB - 1. The effect of the microtubule-disruptive agent, nocodazole (methyl [5-(2 thienylcarbonyl)-1H-benzimidazol-2-yl] carbamate), on the water permeability response to vasopressin or the synthetic cyclic AMP analogue, 8 parachlorophenylthio-cyclic AMP (8-CPT-cAMP), has been investigated in isolated cortical collecting tubules from rabbit kidneys, perfused in vitro. 2. Pre treatment with nocodazole, 1-4 micrograms ml-1, had no significant effect on basal water permeability, but inhibited the increase in hydraulic conductivity elicited by vasopressin, 50 microU ml-1, in a dose-dependent manner. Inhibition of the response to the hormone averaged 65 +/- 6% (n = 8, P less than 0.001) at a nocodazole concentration of 4 micrograms ml-1. 3. Nocodazole, 1-4 micrograms ml 1, had no effect on the increase in lumen-negative potential difference (PD) induced by the hormone. 4. Pre-treatment with nocodazole, 4 micrograms ml-1, inhibited the development of the water permeability response to 8-CPT-cAMP, 1.8 x 10(-5) M, by 45 +/- 7% (n = 7, P less than 0.001). 5. When collecting tubules were exposed to nocodazole, 4 micrograms ml-1, after the hydrosmotic response to vasopressin had been fully established, the drug had no inhibitory effect on the maintenance of a high water permeability. 6. The results are consistent with the view that cytoplasmic microtubules play a role in the initiation of the water permeability response to vasopressin in the mammalian cortical collecting tubule at a cellular site beyond the generation of cyclic AMP. PMID- 2559201 TI - Fatty-acid spin probe interactions with erythrocyte ghosts and liposomes prepared from erythrocyte ghosts. AB - A model for the binding of 5-nitroxide stearate, I(12.3), to human erythrocyte ghosts was developed by comparing spin probe interactions with ghosts and liposomes prepared from ghosts. At low probe/lipid (P/L less than 1/2500), I(12.3) binds to a similar class of high-affinity, noninteracting sites in both ghosts and liposomes, indicating that lipid moieties are responsible for probe uptake. Saturation occurs in both systems with increasing P/L, and, at higher loading (e.g., P/L = 1/360 for ghosts and liposomes), the probe inserts itself at initially dilute sites to form a class of low-affinity sites consisting of clusters of variable size. At still higher P/L ranges (greater than 1/100), much increased probe uptake was observed in ghosts than in liposomes, which was attributed to another class of low-affinity sites, representing nonspecific interactions of I(12.3) with membrane proteins. The nature of the spectral components and ultrafiltration experiments with ghosts labeled at high P/L indicate that both 'dilute' and 'clustered' I(12.3) are due to membrane incorporated probe. PMID- 2559200 TI - Antipeptide antibodies to the beta 2-adrenergic receptor confirm the extracellular orientation of the amino-terminus and the putative first extracellular loop. AB - We developed site-directed rabbit antisera against synthetic peptides selected from the deduced amino acid sequence of the hamster lung beta 2-adrenergic receptor (amino acids 16-31 and 174-189, respectively). All antisera directed against peptide 1 (four of four rabbits) as well as two antisera directed against peptide 2 (two of four rabbits) recognized the purified beta 2-adrenergic receptor in immunoblot conditions when used at a dilution of 1:500. Antisera directed against peptide 1 as well as peptide 2 were able to immunoprecipitate iodinated as well as 125I-cyanopindolol labeled beta 2-adrenergic receptor. This last result implies that the recognized epitopes do not contain the 125I cyanopindolol binding domain of the beta 2-adrenergic receptor. Immunoblot experiments performed on membrane fractions from hamster lung tissue showed that immunoreactive bands at 64,000, 57,000, 47,000, 44,000 and 38,000 daltons were specifically detected. When purified beta 2-adrenergic receptor was iodinated and submitted to glycolytic and/or tryptic treatments, species with similar molecular weights could be recovered. Then, the immunoreactive bands probably correspond to native beta 2-adrenergic receptor and to degradative or nonglycosylated species of this molecule. The antisera were also able to detect immunoreactive molecules in murine and human cell lines, suggesting conservation of the probed sequences between these species. Enzymatic linked immunosorbent assay tests on intact cells and immunofluorescence studies confirmed that the amino-terminus and putative first extracellular loop are extracellularly located. Immunofluorescence studies on mouse brain primary cultures showed that cells expressing beta 2-adrenergic receptor-like molecules exhibited a neuronal phenotype. PMID- 2559202 TI - Activation and conductance properties of ryanodine-sensitive calcium channels from brain microsomal membranes incorporated into planar lipid bilayers. AB - Rat brain microsomal membranes were found to contain high-affinity binding sites for the alkaloid ryanodine (kd 3 nM, Bmax 0.6 pmol per mg protein). Exposure of planar lipid bilayers to microsomal membrane vesicles resulted in the incorporation, apparently by bilayer-vesicle fusion, of at least two types of ion channel. These were selective for Cl- and Ca2+, respectively. The reconstituted Ca2+ channels were functionally modified by 1 microM ryanodine, which induced a nearly permanently open subconductance state. Unmodified Ca2+ channels had a slope conductance of almost 100 pS in 54 mM CaHEPES and a Ca2+/TRIS+ permeability ratio of 11.0. They also conducted other divalent cations (Ba2+ greater than Ca2+ greater than Sr2+ greater than Mg2+) and were markedly activated by ATP and its nonhydrolysable derivative AMP-PCP (1 mM). Inositol 1,4,5-trisphosphate (1-10 microM) partially activated the same channels by increasing their opening rate. Brain microsomes therefore contain ryanodine-sensitive Ca2+ channels, sharing some of the characteristics of Ca2+ channels from striated but not smooth muscle sarcoplasmic reticulum. Evidence is presented to suggest they were incorporated into bilayers following the fusion of endoplasmic reticulum membrane vesicles, and their sensitivity to inositol trisphosphate may be consistent with a role in Ca2+ release from internal membrane stores. PMID- 2559203 TI - Agonist-induced activation of Na+/H+ exchange in rat parotid acinar cells. AB - The present studies were designed to test our previous suggestion that Na+/H+ exchange was activated by muscarinic stimulation of rat parotid acinar cells. Consistent with this hypothesis, we demonstrate here that intact rat parotid acini stimulated with the muscarinic agonist carbachol in HCO3- -free medium show an enhanced recovery from an acute acid load as compared to similarly challenged untreated preparations. Amiloride-sensitive 22Na uptake, due to Na+/H+ exchange, was also studied in plasma membrane vesicles prepared from rat parotid acini pretreated with carbachol. This uptake was stimulated two-fold relative to that observed in vesicles from control (untreated) acini. This stimulation was time dependent, requiring approximately 15 min of acinar incubation with carbachol to reach completion, and was blocked by the presence of the muscarinic antagonist atropine (2 x 10(-5) M) in the pretreatment medium. The effect of carbachol was dose dependent with K0.5 approximately 3 x 10(-6) M. Stimulation of the exchanger was also seen in vesicles prepared from acini pretreated with the alpha adrenergic agonist epinephrine, but not with the beta-adrenergic agonist isoproterenol, or with substance P. Kinetic analysis indicated that the stimulation induced by carbachol was due to an alkaline shift in the pH responsiveness of the exchanger in addition to an increased apparent transport capacity. Taken together with previous results from this and other laboratories, these results strongly suggest that the Na+/H+ exchanger and its regulation are intimately involved in the fluid-secretory response of the rat parotid. PMID- 2559204 TI - Calpain I activates Ca2+ transport by the reconstituted erythrocyte Ca2+ pump. AB - Calpain I purified from human erythrocyte cytosol activates both the ATP hydrolytic activity and the ATP-dependent Ca2+ transport function of the Ca2(+) translocating ATPase solubilized and purified from the plasma membrane of human erythrocytes and reconstituted into phosphatidylcholine vesicles. Following partial proteolysis of the enzyme by calpain I, both the initial rates of calcium ion uptake and ATP hydrolysis were increased to near maximal levels similar to those obtained upon addition of calmodulin. The proteolytic activation resulted in the loss of further stimulation of the rates of Ca2+ translocation or ATP hydrolysis by calmodulin as well as an increase of the affinity of the enzyme for calcium ion. However, the mechanistic Ca2+/ATP stoichiometric ratio was not affected by the proteolytic treatment of the reconstituted Ca2(+)-translocating ATPase. The proteolytic activation of the ATP hydrolytic activity of the reconstituted enzyme could be largely prevented by calmodulin. Different patterns of proteolysis were obtained in the absence or in the presence of calmodulin during calpain treatment: the 136-kDa enzyme was transformed mainly into a 124 kDa active ATPase fragment in the absence of calmodulin, whereas a 127-kDa active ATPase fragment was formed in the presence of calmodulin. This study shows that calpain I irreversibly activates the Ca2+ translocation function of the Ca2(+) ATPase in reconstituted proteoliposomes by producing a calmodulin-independent active enzyme fragment, while calmodulin antagonizes this activating effect by protecting the calmodulin-binding domain against proteolytic cleavage by calpain. PMID- 2559205 TI - Temperature dependence of gating current in myelinated nerve fibers. AB - Asymmetrical displacement currents and Na currents of single myelinated nerve fibers of Xenopus laevis were studied in the temperature range from 5 to 24 degrees C. The time constant of the on-response at E = 4 mV, tau on, was strongly temperature dependent, whereas the amount of displaced charge at E = 39 mV, Qon, was only slightly temperature dependent. The mean Q10 for tau on-1 was 2.54, the mean Q10 for Qon was 1.07. The time constant of charge immobilization, tau i, at E = 4 mV varied significantly (alpha = 0.001) with temperature. The mean Q10 for tau i-1 was 2.71 +/- 0.38. The time constants of immobilization of gating charge and of fast inactivation of Na permeability were similar in the temperature range from 6 to 22 degrees C. The Qoff/Qon ratio for E = 4 mV pulses of 0.5 msec duration decreased with increasing temperature. The temperature dependence of the time constant of the off-response could not be described by a single Q10 value, since the Q10 depended on the duration of the test pulse. Increasing temperature shifted Qon (E) curves to more negative potentials by 0.51 mV K-1, but shifted PNa (E) curves and h infinity (E) curves to more positive potentials by 0.43 and 0.57 mV K-1, respectively. h infinity (E = -70 mV) increased monotonously with increasing temperature. The present data indicate that considerable entropy changes may occur when the Na channel molecule passes from closed through open to inactivated states. PMID- 2559206 TI - Specificity and mechanism of antitermination by Q proteins of bacteriophages lambda and 82. AB - Lambdoid phage late gene operons are positively regulated by genome-specific antiterminator proteins encoded by the Q gene of each phage. In this paper, we compare the activity of phage lambda and phage 82 Q proteins. Q82-mediated antitermination, like that of Q lambda, involves a transcription pause during which the regulator can modify RNA polymerase. We show that the activities of both Q82 and Q lambda are genome-specific in chasing RNA polymerase out of the early pause sites and in mediating antitermination. Finally, we show that the length of the RNA in the paused complex, or the exact position of the pause, affects the efficiency with which Q82 chases RNA polymerase out of the pause. PMID- 2559208 TI - Activation of Chi recombinational hotspots by RecBCD-like enzymes from enteric bacteria. AB - Chi sites, 5'G-C-T-G-G-T-G-G-3', enhance homologous recombination in Escherichia coli and are activated by the RecBCD enzyme. To test the ability of Chi to be activated by analogous enzymes from other bacteria, we cloned recBCD-like genes from diverse bacteria into an E. coli recBCD deletion mutant. Clones from seven species of enteric bacteria conferred to this deletion mutant recombination proficiency, Chi hotspot activity in lambda Red- Gam- vegetative crosses, and RecBCD enzyme activities, including Chi-dependent DNA strand cleavage. Three clones from Pseudomonas aeruginosa and Ps. putida conferred recombination proficiency and ATP-dependent nuclease activity, but neither Chi hotspot activity nor Chi-dependent DNA cleavage. These results imply that Chi has been conserved as a recombination-promoting signal for RecBCD-like enzymes in enteric bacteria but not in more distantly related bacteria such as Pseudomonas spp. We discuss the possibility that other, presently unknown, nucleotide sequences serve the same function as Chi in Pseudomonas spp. PMID- 2559209 TI - Tetragonal crystals of canine myeloperoxidase suitable for X-ray structural analysis. AB - A new tetragonal crystal form of canine myeloperoxidase grown by precipitation with ammonium sulfate is described. The space group is P4(1)2(1)2 or P4(3)2(1)2 with unit cell dimensions. a = b = 133.0 A, c = 203.6 A, and a single molecule in the asymmetric unit. The crystals diffract to Bragg spacings of 2.5 A and are suitable for a medium-resolution structure determination. PMID- 2559207 TI - Sequences required for antitermination by phage 82 Q protein. AB - The gene Q antiterminator proteins of phages lambda and 82 modify RNA polymerase at sites (named qut) that are close to, and apparently inseparable from the promoters themselves. Modification occurs while RNA polymerase has paused close to the start site, at nucleotide 16 for lambda, and nucleotides 15 and 25 for phage 82. We present a deletion analysis of the phage 82 qut site that identifies sequences required for pausing and shows that these sequences also are required for efficient Q function in vivo and in vitro. We show (1) that deletions as close as +5 to the RNA start site retain some ability to be modified by Q82, suggesting that part of the qut site is in the non-transcribed region of the promoter; (2) that NusA protein is required for activity of Q82 on certain qut82 site deletions, whereas it only modestly stimulates antitermination from the native qut82 site; and (3) that qut82 is active only on RNA polymerase that initiates at the qut-associated promoter, and not on RNA polymerase that initiates upstream and passes through an otherwise active qut82 site. PMID- 2559210 TI - Nonuniform evolution of duplicated, developmentally controlled chorion genes in a silkmoth. AB - We report the sequence of A/B.L1, a tightly linked pair of genes from the A and B chorion families in Bombyx mori. Comparison with the previously characterized A/B.L11 and A/B.L12 pairs from the same species reveals moderate sequence divergence, which is clearly nonuniform. Although the average divergence of A/B.L12 from the other two pairs is more than double that between A/B.L11 and A/B.L1, the ratio differs by more than 30-fold in different DNA regions. One domain of the A gene is highly divergent between A/B.L12 and A/B.L1 or A/B.L11, but essentially invariable in the latter two. In well-aligned DNA segments, nearly all mutated sites (111/112) show variants shared by two of the three sequences, in 42% of the cases between the more distantly related pairs (A/B.L12 and either A/B.L1 or A/B.L11). Eight of the variants shared by distantly related pairs are clustered within 51 bp, suggesting the possibility that they arose through sequence transfers between gene pairs, rather than being primitive or resulting from independent mutations. The short intergenic, putatively regulatory DNA of the developmentally middle A/B.L1 and A/B.L11 pairs resembles that of the late HcA/HcB pairs, particularly in patches that may correspond to cis-regulatory elements. PMID- 2559211 TI - Transgenic regulation of moth chorion gene promoters in Drosophila: tissue, temporal, and quantitative control of four bidirectional promoters. AB - Bidirectional chorion gene promoter regions from three silkmoth species, Bombyx mori, Antheraea pernyi, or Antheraea polyphemus (members of two different moth families), were tested for their ability to transcriptionally activate a bacterial marker gene (chloramphenicol acetyltransferase) in transformant Drosophila. Relatively short 5' flanking DNA fragments (272-367 bp) of chorion gene pairs are sufficient to confer a high degree of tissue and choriogenic stage specificity of expression to the marker gene. Thus, significant conservation of molecular interactions controlling transcription during choriogenesis is observed between the distantly related orders, Lepidoptera and Diptera. However, quantitative and fine temporal regulation in the Drosophila host does not fully parallel the in situ regulation in moths, indicating that some regulatory protein DNA interactions have diversified in the approximately 250 million years since the last common ancestor of these insect groups. Limited in vitro mutagenesis of a B. mori promoter DNA has shown that a central 189-bp region includes elements sufficient for the qualitative specificity of chorion-specific expression. The same experiments have shown that a previously identified essential element, centered on the TCACGT hexamer, is not sufficient for chorion-specific expression: an additional essential element or elements are found farther upstream, within a 112-bp DNA region. Comparisons of silkmoth and Drosophila chorion gene promoter sequences have identified some candidates for cis-acting elements involved in the developmental regulation of chorion gene expression. PMID- 2559214 TI - [Molecular pathology of HBV induced hepatocarcinogenesis]. PMID- 2559213 TI - Central and peripheral beta-adrenergic influences on reticulo-rumen and upper-gut myoelectrical activity in sheep. AB - The effects of intravenous (i.v.) and intracerebroventricular (i.c.v.) administration of beta-adrenoceptor agonists were evaluated on the reticulo-rumen and upper-gut myoelectrical activity in six ewes chronically fitted with intraparietal electrodes and a cannula in a lateral ventricle of the brain. Intravenous infusion of the beta 1 agonist dobutamine (30 micrograms/kg/min for 15 min) reduced the frequency of reticulo-ruminal and abomasal contractions and stimulated duodeno-jejunal motility, inducing a Phase III on the jejunum. These effects were reproduced by i.c.v. dobutamine at a dose of 10 micrograms/kg. Intravenous infusion of the beta 2 agonist ritodrine (15 micrograms/kg/min for 15 min) selectively inhibited antral and duodenal motility. Ritodrine i.c.v. (15 micrograms/kg) did not affect forestomach or gastrointestinal motility. The mixed beta 1, beta 2 agonist isoprenaline infused i.v. (0.6 micrograms/kg/min for 15 min) reproduced the effects of i.v. dobutamine, except at the antro-duodenal level which was strongly inhibited. The effects of i.v. dobutamine were antagonized by i.v. or i.c.v. acebutolol, a specific beta 1 antagonist. The effects of i.v. ritodrine were blocked by i.v. but not i.c.v. administration of propranolol, a mixed beta 1, beta 2 antagonist. These data indicate that the stimulation of central beta 1 adrenoceptors inhibits forestomach and antral motility and stimulates duodeno-jejunal motility. Stimulation of peripheral beta 2 adrenoceptors selectively inhibits duodeno-jejunal motility. PMID- 2559212 TI - Effects of the N-methyl-D-aspartate receptor blocker MK-801 on neurologic function after experimental brain injury. AB - Pharmacologic inhibition of excitatory amino acid (EAA) neurotransmission attenuates cell death in models of global and focal ischemia and hypoglycemia and improves neurologic outcome after experimental traumatic spinal cord injury. The present study examined the effects of the noncompetitive N-methyl-D-aspartate (NMDA) receptor blocker MK-801 on cardiovascular and neurologic function after experimental fluid-percussion (FP) brain injury in the rat. Animals received either an intravenous bolus of MK-801 (1 mg/kg) or saline (equal volume) 15 min prior to FP brain injury or 15 min following FP brain injury. MK-801 pretreatment significantly improved postinjury cardiovascular variables and attenuated postinjury neurologic dysfunction. Postinjury treatment with MK-801 also significantly improved cardiovascular variables, but had little effect on postinjury neurologic scores. These results suggest that EAA neurotransmitters may be involved in the pathophysiological sequelae of traumatic brain injury and that noncompetitive blockade of the NMDA receptor prior to brain injury may reduce EAA-induced damage and limit neurologic dysfunction. PMID- 2559215 TI - [A case of herpes zoster duplex with generalized eruption--study of the pathogenesis of the eruptions]. AB - A 77-year-old man who developed herpes zoster duplex with generalized eruptions is presented. Three strains of varicella-zoster virus (VZV) were isolated from 2 skin lesions of herpes zoster and one of the generalized eruptions. For strain differentiation, these 3 strains were investigated by restriction endonuclease analysis of the viral DNA. The 3 strains were identical, suggested that a single strain of VZV had been residing in a latent form in multiple spinal ganglions of this patient. We speculate that this strain of VZV in remote ganglions was reactivated by the operation of gastric cancer as a trigger, and, as a result, the 2 skin lesions of herpes zoster appeared, followed by development of generalized eruptions due to virus propagation from zoster lesions. PMID- 2559216 TI - [Two cases of oral leukoplakia possibly associated with human papillomavirus]. AB - Two cases of oral leukoplakia were studied immunohistologically and ultrastructurally. By means of avidin-biotin peroxidase complex technique using papillomavirus specific antibody, distinct nuclear staining of epithelial cells of middle and upper layers was observed in both cases. Further evidence for the presence of human papillomavirus was obtained by the electron micrograph, which revealed virus-like particles (30-40 nm in diameter) within the nucleus. PMID- 2559217 TI - Potential usefulness of a cultured glioma cell line induced by Rous sarcoma virus in B10.A mouse as an immunotherapy model. AB - A cultured glioma cell line, SR-B10.A, which was derived from a brain tumor induced in an adult female B10.A mouse by Rous sarcoma virus (RSV), has been established. The morphological appearance of the tumor produced by s.c. inoculating SR-B10.A cells was analogous to an astrocytoma of human glioma. Glial fibrillary acidic protein as well as S-100 protein was positive in these SR-B10.A tumor cells. A population doubling time of the cultured cells was 18.5 hours. Chromosomal analysis revealed a defect in one of the sex chromosomes. Integration of RSV genome was proven to be positive in SR-B10.A cells. It was possible to generate cytotoxic effector cells in the syngeneic B10.A mouse against SR-B10.A. The tumor-bearing syngeneic hosts harbored a suppressor activity in the splenocytes. Although recombinant human tumor necrosis factor (rH-TNF) had no growth inhibitory effect on the SR-B10.A cells in vitro, the s.c. implanted and growing tumor regressed when rH-TNF was administered intratumorally several times. In addition, this anti-tumor effect was completely abrogated when the host mice were treated with wholebody x-ray irradiation prior to the tumor cells inoculation. In contrast, neither rH-TNF (i.v.) nor cyclophosphamide (i.p.) induced the regression of SR-B10.A, indicating that efficacy of the locally administered rH-TNF is dependent on the host immune mechanism. These results suggest that SR-B10.A is a potentially useful tumor model in evaluating efficacy of immunomodulators. PMID- 2559218 TI - Thermal injury-induced non-specific resistance to fatal Pseudomonas aeruginosa burn-infection in mice. AB - Nonlethal thermal injury in mice results in rapid death by immediate injection of 10(3) viable P. aeruginosa in the skin of the burn sites. Resistance to the lethal burn combined with P. aeruginosa infection developed 24 h after initial thermal injury and reached maximal effect 7 days later; it then continued for at least 21 days. The optimal survival was achieved when the first thermal injury was made for 7 seconds at 350 degrees C. Increased resistance, but for a short period could also be obtained by injection of lipopolysaccharide (LPS) 1-4 days prior to the burn-P. aeruginosa infection. However, when the LPS was injected immediately after the burn-infection, the lethal effect was increased. The induction of late protection after thermal injury and bacterial infection was demonstrated with P. aeruginosa organisms only. Under similar schedule of thermal injury resistance was not induced by infection with Semliki forest virus. On the contrary viral infection increased the susceptibility of burned mice to a fatal outcome. Immune or natural antibodies were not elevated in the sera of post burn mice. Furthermore, delayed type hypersensitivity response, as evaluated by a footpad weight assay was inhibited and this inhibition persisted at least for 7 days post burn. The thymus weight and its lymphoid cell content in thermally injured mice decreased significantly 7 days post burn, whereas the weight of the spleen increased and it contained fewer lymphocytes per gram tissue. We suggest that endotoxin entering the systemic circulation post-burn might be one of the factors contributing to the early sensitivity and the late protection against the fatal P. aeruginosa burn-infection. PMID- 2559219 TI - Ultrastructural observations on damage to schistosomula of Schistosoma japonicum by mouse neutrophils in vitro. AB - Ultrastructure of schistosomula of Schistosoma japonicum was studied in relation to the effect of mouse neutrophils in vitro. In the presence of antibody and complement, damage to the schistosomular tegument covered with many neutrophils began to appear within 1 h incubation. Exfoliation of the granular cytoplasm of the tegument, disorganization of the muscle layers and vacuolation in the inner tissues were seen in a part of neutrophil-attached parasites by 2 h incubation. However, the majority of schistosomula with fewer cells retained their integrity till 16 h. The cytochemical examination of schistosomula incubated with mouse neutrophils for 1 and 2 h demonstrated clearly the localization of the peroxidase activity on the surfaces of the cells and the parasites. PMID- 2559220 TI - [A study of 75 untreated lung cancers in the elderly]. AB - The records of 1280 patients autopsied at the Yokufukai Geriatric Hospital from October 1, 1973 to August 31, 1987 were reviewed and 75 patients with untreated lung cancer, aged 70 or older, were selected. The mean age and standard deviation was 82.1 +/- 5.4 years. Male consisted of 34 subjects and 41 were female of. Histological study revealed 42 cases of adenocarcinoma, 19 cases of squamous cell carcinoma, 7 cases of small cell carcinoma, 2 cases of large cell carcinoma, 1 case of carcinoid and 4 cases of the other types. The mean survival period of 44 untreated patients diagnosed as lung cancer during life was 21.1 +/- 24.1 months. The mean survival periods for 24 patients with adenocarcinoma and 11 patients with squamous cell carcinoma were 24.0 +/- 29.3 and 12.9 +/- 11.7 months, respectively. There was no statistically significant difference in the mean survival period of adenocarcinoma and squamous cell carcinoma. 9% of 44 untreated lung cancers survived for at least 5 years, although the survival rate was slightly lower than that generally reported in the literature. On the basis of staging of TNM classification at the autopsy, the mean survival period from the diagnosis for 13 patients with stage 1 and 27 patients with stage 4 were 27.5 +/- 33.3 and 18.5 +/- 19.7 months, respectively. The incidence of brain metastasis in 75 cases was 14.7%. In this study, adenocarcinoma was more predominant in the elderly (56.0%). An inverse relationship of age to stage was partially observed. PMID- 2559221 TI - [Dilated cardiomyopathy associated with tumor pulmonary microembolism of small cell lung cancer, an elderly case report]. PMID- 2559222 TI - [Changes in cyclic-AMP levels in the brain of rat administered methylmercury chloride]. PMID- 2559223 TI - Comparison of Mg, Mn, and Co ions affecting the beta-adrenoceptor-mediated membrane response in the guinea-pig taenia caeci. AB - Electrical activity was recorded intracellularly from the muscle cell of guinea pig taenia caeci in Locke solution. Membrane potential was -46.4 mV, and spike potentials were discharged spontaneously. Isoprenaline (3 microM) hyperpolarized the membrane and suppressed the spike discharge. The hyperpolarization by isoprenaline was increased at low K (2 mM), while decreased at high K (11.8, 29.5, 59 mM). The hyperpolarization by isoprenaline was potentiated in the presence of external Mg ions, depending on the concentration of Mg (0-9.6 mM). Forskolin (3 microM) and papaverine (30 microM) hyperpolarized the membrane; the effects were augmented by 1.2 mM Mg. The hyperpolarization in response to 3 microM isoprenaline or 100 microM papaverine was inhibited by Mn, Co, and low Ca (1 mM) whereas it was not affected by high Ca (7.5 mM). Verapamil (0.5, 2 microM) had no influence of the hyperpolarization caused by isoprenaline. It was discussed that extracellular and/or intracellular Mg and Ca ions played important roles in the beta-adrenoceptor-mediated action on the smooth muscle membrane of taenia caeci. PMID- 2559224 TI - A voltage-dependent calcium current in mouse MC3T3-E1 osteogenic cells. AB - MC3T3-E1 osteogenic cells in a growing state were voltage-clamped by the whole cell patch-clamp method. The MC3T3-E1 cells exhibited a transient, fast inactivating Ca inward current upon depolarizing pulses from a holding potential of -80 mV. This current had a threshold of activation of about -50 mV and was insensitive to the dihydropyridine, nifedipine. These results show that MC3T3-E1 cells have a voltage-dependent Ca channel corresponding to the "T-type." PMID- 2559225 TI - Effects of KT-362, a new Na and Ca influx and Ca release inhibitor, on canine ventricular arrhythmias. AB - Antiarrhythmic effects of the new drug KT-362, which was reported to suppress Na and Ca currents of cardiac cells and also to suppress intracellular Ca release in isolated smooth muscle preparations, were examined using two-stage coronary ligation-, digitalis- and adrenaline-induced ventricular arrhythmias in the dog. Intravenous KT-362 at 10 mg/kg suppressed coronary ligation arrhythmia both at 24 and 48 hr after ligation, and the minimum effective plasma concentrations for arrhythmias induced by 24 hr coronary ligation and 48 hr coronary ligation were 6.1 +/- 1.7 and 8.6 +/- 2.7 micrograms/ml, respectively. Antiarrhythmic effects were accompanied by transient hypotension. Oral administration of 70-100 mg/kg was also effective on 24 hr coronary ligation arrhythmia. However, there was no prominent hypotension in these experiments. Intravenous KT-362 at 3 mg/kg suppressed digitalis arrhythmia; and the minimum effective plasma concentration was 3.3 +/- 1.2 micrograms/ml, which was lower than the effective plasma concentrations for coronary ligation arrhythmias. Intravenous KT-362 at 1 mg/kg also suppressed adrenaline arrhythmia; and the minimum effective plasma concentration was 1.0 +/- 0.1 microgram/ml, the lowest among the effective plasma concentrations. These pharmacological profiles of KT-362 are quite different from those of class 4 Ca antagonists, but similar to those of class 1 drugs such as propafenone. Though KT-362 has a hypotensive effect, it is effective on canine ventricular arrhythmias; thus its clinical usefulness for supraventricular and ventricular arrhythmias is expected. PMID- 2559226 TI - [Serum antibody titers against various viruses in idiopathic interstitial pneumonia]. AB - As the first step to investigate a possibility that viral infection is involved in the pathogenesis of idiopathic interstitial pneumonia (IIP), serum antibody titers against various viruses were studied in 98 IIP patients. The patients were positive for herpes simplex virus, cytomegalovirus, rubella virus, parainfluenza virus 3, EBV-VCA IgG and adeno virus 1, which were also assessed in 45 normal subjects. As results, IIP patients showed higher titers of EBV-VCA IgG (p less than 0.01). When the frequency of positive populations for each of the 6 viral antibodies was calculated, the values of the frequency in all the 6 viruses were found to be very high in both patients and normal subjects, and there was no significant difference between IIP patients and normal controls. In conclusion, a specific relationship between IIP and a certain viral infection was not observed by studying serum antibody titers against various viruses in IIP patients. It might be worthwhile studying why EBV-VCA IgG titers were shown to be significantly higher in IIP patients than in normals. PMID- 2559228 TI - [A case of Recklinghausen's disease associated with multiple emphysematous bullae and lung cancer]. AB - A 36-year-old man was admitted with cough and sputum. He had cafe-au-lait spots and multiple subcutaneous neurofibromas. Chest X-ray revealed multiple emphysematous bullae in bilateral upper lobes and a tumor in the bulla of the right upper lobe. Needle aspiration biopsy of the tumor showed small cell carcinoma. Although chemotherapy and radiation resulted in decrease in tumor size, it subsequently increased in size and he died 11 months after admission. Including this case there have been 7 reports of Recklinghausen's disease associated with multiple lung cysts and 8 reports with lung cancer in Japan. However, cases with lung cysts and cancer are very rare. The cancer of this case was considered to be associated with emphysematous bulla rather than Recklinghausen's diseases. PMID- 2559227 TI - [A pathological study of carcinoma of the lung and pneumoconiosis]. AB - Pathological studies on lung carcinoma associated with pneumoconiosis were made in order to clarify the etiological significance of pneumoconiosis in carcinoma of the lung. Between 1960 and 1986, the author evaluated approximately 450 autopsies of pneumoconiosis. Of these, 150 were consecutive autopsies in our over laboratory and the remaining 300 were kindly provided by other hospitals. Carcinoma of the lung was seen in 48 of the autopsies. Pathological studies determine histologic type, cancer site, and severity of pneumoconiosis were performed in the 48 cases. The severity of pneumoconiosis was determined by the extent of progressive massive fibrosis (PMF). Simple pneumoconiosis was classified as mild, pneumoconiosis with PMF smaller than segmental region as moderate and pneumoconiosis with PMF larger than lung segment as severe. Carcinoma of the lung was seen in 25 of our own consecutive autopsies, an incidence of 17.9%. The incidence of lung carcinoma was high among cases of mild pneumoconiosis and low among cases of severe pneumoconiosis. Overall, the predominant cancer was squamous cell carcinoma (54.2%) followed by small-cell carcinoma (22.9%) and adenocarcinoma (14.6%). There was a clear trend that most of the squamous cell carcinomas were found in the larger airways, whereas the adenocarcinoma was found only in the peripheral lung tissues. In cases of mild pneumoconiosis, the majority of tumors arose in the right, upper and larger airways. On the other hand, in cases of moderate and severe pneumoconiosis, more tumors arose in the left, lower and peripheral lung areas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559229 TI - [A case report of large cell lung cancer detected in the course of antituberculous therapy after a giant bullectomy]. AB - A case of a 60-year-old who male developed lung cancer after surgical resection of a emphysematous giant bulla. At first, he showed a emphysematous giant bulla associated with infected bulla in the right lung on the chest roentgenograph. After a giant bullectomy the resected specimen, suggested possible pulmonary tuberculosis. Thereafter, he was followed for one year with antituberculous therapy. Nevertheless, the chest roentgenograph taken one year after surgery, showed a new tumor density (1.5 x 2.0 cm) in the right lung. Due to the rapid growth of the lung tumor, right upper lobectomy was performed. The histological diagnosis was a large cell lung cancer (giant cell type). The postoperative staging of the tumor proved to be T2N0M0. He was treated with systemic chemotherapy of CDDP 120 mg, VDS 4.5 mg, and MMC 12 mg. Unfortunately, he died of cancer recurrence 5 months after lobectomy. Clinical evaluation of the emphysematous giant bulla associated with lung cancer was performed. PMID- 2559230 TI - [Pleomorphic adenoma of the lung]. AB - A 41 year-old male was admitted to our hospital, because a suspicious shadow in the right lower lung field was on X-ray. Investigations could not confirm cancer but were highly suggestive of it. Finally, right lower lobectomy was performed, and the histopathological finding of the tumor resected were a mixture of epithelial components in myxomatous tissue, so a diagnosis of pleomorphic adenoma of the lung was made. PMID- 2559231 TI - Cytotoxic effect of acyclovir on cultured mammalian cells to which herpesvirus thymidine kinase gene was introduced. AB - The cytotoxic effects of acyclovir, which is a purine nucleoside analogue and is known as an antibiotic substance, were examined on three lines of rat skin fibroblast FR cells; normal FR cells, FRtk- cells which are deficient in the activity of thymidine kinase (tk) and FRtk-HSVtk+ cells which were prepared by introducing herpes simplex virus' tk gene to FRtk- cells. When FRtk-HSVtk+ cells growing exponentially were incubated in the presence of acyclovir for 4 h, the surviving fractions of the cells decreased in a concentration-dependent manner. Whereas, decrease of the surviving fractions was almost indiscernible in both FR cells and FRtk- cells at the whole ranges of drug-concentrations tested. These results indicate that acyclovir is phosphorylated by the herpes simplex virus' tk and becomes toxic to FRtk-HSVtk+ cells. This also means that FRtk-HSVtk+ cells are useful for the investigation of the biological activity of nucleoside analogues. PMID- 2559232 TI - [Dilated cardiomyopathy: sequela of viral myocarditis?]. AB - The study was undertaken to examine 100 patients with dilated cardiomyopathy (DCM) and 62 with viral myocarditis (VM). Serological studies revealed that 60% of the VM patients and 25% of the DCM patients had diagnostically significant antibody titers to Coxsackie B virus. The patients from the two groups displayed NK cell deficiency, quantitative and functional suppressor defect, and severe derangements in the system of interferons. Unlike DCM patients, some VM patients showed greater quantities of cells having phenotype OKT 8. A follow-up demonstrated that 11% of the VM patients developed a symptom complex typical of DCM. Changes in immunological parameters appeared as smaller numbers of the cells with phenotype T 8 in the patients. Immunogenetic study revealed the genetic marker HLA-B12 common in VM and DCM. VM evolved into DCM in 75% of the VM patients who had antigen B12. Clinical, serological, and immunological studies enabled a risk group of patients with VM in whom the disease might develop to DCM to be detected. PMID- 2559233 TI - Renal medullary Na-K-ATPase and hypoxic injury in perfused rat kidneys. AB - We wished to see if chronic alterations in Na-K-ATPase activity in the medullary thick ascending limb would modify the susceptibility of its cells to the hypoxic injury produced by perfusion of the isolated kidney. Rats were fed a diet high (64%) or low (8%) in protein for three weeks. Renal medullary Na-K-ATPase was 75 +/- 12 U/mg protein/hr (mean +/- SE) in the high protein group and 44 +/- 3 in rats given low protein. After 90 minutes of perfusion, the kidneys of rats fed a high protein diet showed almost all mTAL cells near the inner medulla with severe damage (93 +/- 4.8%), whereas the same zone in perfused kidneys of rats on a low protein diet showed only 47 +/- 7.7% injury. In a similar fashion, damage to mTAL cells seen in perfused kidneys was greatly augmented by compensatory renal hypertrophy produced by removal of the contralateral kidney two weeks earlier, and by a diet high in potassium given for two weeks, procedures which also increased the activity of medullary Na-K-ATPase. The results suggest that the level of transport work of medullary cells mediated by Na-K-ATPase is a determinant of the vulnerability of mTAL cells to hypoxic injury. PMID- 2559234 TI - Correction of glucose intolerance and the impaired insulin release of chronic renal failure by verapamil. AB - Insulin release from pancreatic islets is impaired in chronic renal failure (CRF), and this is due to the state of secondary hyperparathyroidism of CRF. This defect in association with resistance to the peripheral action of insulin-caused glucose intolerance in CRF. It has been suggested that the impaired insulin release induced by excess parathyroid hormone (PTH) is related to the ability of the hormone to augment calcium entry into the pancreatic islets, resulting in accumulation of calcium in the pancreas. Therefore, a calcium channel blocker may antagonize this effect of PTH, and hence normalize glucose tolerance in CRF. The present study examined this postulate by studying intravenous glucose tolerance and insulin release from pancreatic islets in normal and CRF rats and in CRF animals treated with the calcium channel blocker, verapamil. Rats with 42 days of CRF displayed impaired glucose tolerance, significant reduction (P less than 0.01) in insulin release by islets, and doubling of calcium content of the pancreas (P less than 0.01) as compared to normal rats. Simultaneous treatment of CRF rats with verapamil for 42 days resulted in normal glucose tolerance, higher blood insulin levels during glucose infusion, normal calcium content of the pancreas, and normal insulin secretion by the islets. Treatment of normal rats with verapamil for 42 days did not affect any of the parameters studied. The results show that the calcium channel blocker, verapamil, by preventing calcium accumulation in the pancreas, reversed the abnormalities in insulin release that occur in CRF. This effect allowed a greater rise in blood levels of insulin during glucose infusion in CRF rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559235 TI - Human mesangial cells secrete a GBM-degrading neutral proteinase and a specific inhibitor. AB - The factors operative during normal and disease states which govern the degradation of the glomerular extracellular matrix (ECM, glomerular basement membrane and mesangial matrix) remain poorly understood. These structures consist primarily of types IV and V collagen, which are highly resistant to the action of classical interstitial collagenases. Here we report that an intrinsic glomerular cell type, the human mesangial cell, secretes: 1) a neutral proteinase with specific activity against type IV basement membrane collagen and, 2) an inhibitory protein with the characteristics of the tissue inhibitor of metalloproteinases (TIMP). The purified enzyme has a neutral pH optimum, consists of a basic (pI 8.4) protein doublet with molecular weight of 66 and 68 kilodaltons, and is inhibited by calcium and zinc chelators. The enzyme degrades both soluble and basement membrane type IV collagen, but does not specifically degrade types I or V collagen or casein. Immunohistochemistry of cultured cells demonstrated homogeneous staining for the neutral proteinase antigen, while the TIMP antigen was detected in fewer than one-third of cultured cells. These findings suggest that the synthesis of these proteins may be independently regulated. The secretion of these factors may play an important role in the turnover of the glomerular ECM under basal or pathologic conditions. PMID- 2559236 TI - Effect of converting enzyme inhibition on the course of adriamycin-induced nephropathy. AB - The effect of the converting enzyme inhibitor (CEI) enalapril was assessed in Munich-Wistar rats with established adriamycin nephrosis. Rats were given a single dose of adriamycin and one month later divided into four groups matched for albuminuria, blood pressure, and plasma albumin concentration. Groups 1 and 3 remained untreated while groups 2 and 4 received enalapril. Groups 1 and 2 underwent micropuncture studies after 10 days. These short-term studies showed that enalapril reduced arterial blood pressure (101 +/- 2 vs. 124 +/- 3 mm Hg, group 2 vs. 1, P less than 0.05) and glomerular capillary pressure (54 +/- 1 vs. 61 +/- 2 mm Hg, P less than 0.05) without reducing albuminuria (617 +/- 50 vs. 570 +/- 47 mg/day) or GFR (1.03 +/- 0.04 vs. 1.04 +/- 0.11 ml/min). Groups 3 and 4 were studied at four and at six months to assess the effect of enalapril on progression of renal injury in adriamycin nephrosis. Chronic enalapril treatment reduced blood pressure without reducing albuminuria in group 4. Untreated group 3 rats exhibited a progressive reduction in GFR (0.35 +/- 0.08 ml/min at 4 months, 0.27 +/- 0.07 ml/min at 6 months). Enalapril treatment blunted but did not prevent reduction in GFR in group 4 (0.86 +/- 0.15 ml/min at 4 months, 0.69 +/- 0.13 ml/min at 6 months, both P less than 0.05 vs. group 3). Reduction in GFR was associated with the development of glomerular sclerosis in both treated and untreated rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559237 TI - Rapid hypertonic cell volume regulation in the perfused inner medullary collecting duct. AB - Differential interference contrast microscopic images were used to assess the cell volume regulatory increase (VRI) response of rat IMCD segments isolated from the mid-inner medullary region of pathogen-free Sprague-Dawley rats and perfused in vitro at 37 degrees C. In the absence of ADH. IMCD cells behaved in an osmometric fashion over the range of extracellular osmolalities 290 to 386 mOsm/kg H2O and had an osmotic space equal to 54.2% of total geometric volume. After initial shrinkage in hypertonic perfusing and bathing solutions (340 mOsm/kg H2O using sucrose), cell volume increased rapidly to the isotonic value only in tubules preincubated in ADH (100 microU/ml). The rates of VIR were: ( ADH) 0.0142 +/- 0.0046 nl.min-1.cm-1 or 0.30 +/- 0.10%/min and (+ADH) 0.7225 +/- 0.1278 nl.min-1.cm-1 or 15.42 +/- 2.31%/min (N = 4; P less than 0.01). An overshoot in cell volume was observed on return to isotonic media only in the ADH exposed tubules showing a hypertonic VRI response, indicating that IMCD cells accumulated solute during hypertonic VRI. In the absence of ADH, one mM dibutyryl cyclic AMP mimicked the effect of hormone on hypertonic VRI. This ADH-dependent VRI process required Na+ and (CO2 + HCO3-) in external media and was reduced or abolished by 0.1 mM amiloride, 0.1 mM 4,4'-diisothiocyanatostilbene-2,2-' disulfonic acid (DIDS) in peritubular solutions. These data suggest that ADH dependent, rapid hypertonic cell volume regulation in rat inner medullary collecting duct depends on NA+ uptake, which may be mediated by parallel Na+-H+ and an HCO3(-)-dependent. DIDS-sensitive pathway (such as, Cl+-HCO3- exchanger) in basolateral cell membrane. In addition, a luminal amiloride-sensitive pathway (most likely the cation-selective channel) may contribute to cell volume regulation in the rat IMCD. PMID- 2559238 TI - Epinephrine and dDAVP administration in patients with congenital nephrogenic diabetes insipidus. Evidence for a pre-cyclic AMP V2 receptor defective mechanism. AB - We recently showed that the administration of the antidiuretic V2 specific agonist, 1-desamino[8-D-arginine]vasopressin (dDAVP), to seven male patients with congenital nephrogenic diabetes insipidus (CNDI) did not cause a decrease in blood pressure nor an increase in plasma renin activity or factor VIIIc or von Willebrand factor release. In normal subjects, plasma renin activity, coagulation factors and plasma cyclic AMP are stimulated not only by dDAVP but also by the administration of epinephrine. In the present study, we measured tissue plasminogen activator (activity and antigenicity), von Willebrand factor multimers, plasma and urinary cyclic AMP concentrations following dDAVP or epinephrine administration. We infused epinephrine into three male patients with CNDI. Factor VIIIc and tissue plasminogen activator augmented by 75 to 100% and von Willebrand Factor multimers were increased; plasma renin activity and plasma cyclic AMP concentration increased by 200%. None of these values changed when the same subjects as well as eleven other male patients with CNDI received dDAVP. Furthermore, dDAVP administration increased plasma cyclic AMP concentrations in normal subjects, but not in 14 male patients with CNDI. These results demonstrate the specificity of the extrarenal V2 receptor defect expressed in our patients. The lack of a plasma cyclic AMP response to the administration of dDAVP would suggest an altered pre-cyclic AMP stimulation mechanism. PMID- 2559239 TI - [Use of non-traditional methods of examination in the differential diagnosis of cancer and tuberculosis of the lungs]. AB - The immunoradiometric assay of the blood serum was performed in 61 patient with cancer and in 57 patients with tuberculosis of the lungs to determine the content of adrenocorticotropic hormone (ACTH), hydrocortisone, triiodothyronine++ (T3), thyroxin (T4), ferritin and carcinoembryonic antigen (CEA). In 200 patients (104 suffered from pulmonary tuberculosis, 9--pulmonary cancer), entropy of the protein fractions of the blood serum was determined. It was established that in differential diagnosis of pulmonary cancer and tuberculosis, the determination of CEA, hydrocortisone and entropy of the protein fractions of the blood serum in the complex with the other methods were of significant importance, the determination of ACTH and ferritin content were less important. PMID- 2559240 TI - [Surgical treatment of nonchromaffin paraganglioma]. PMID- 2559241 TI - Beta-adrenergic blood pressure regulation in Shy-Drager syndrome and pheochromocytoma. AB - Both Shy-Drager syndrome and pheochromocytoma are characterized by an abnormal catecholamine secretion, e.g. a reduced secretion in Shy-Drager syndrome, and an excessive stimulation in pheochromocytoma resulting in adrenergic dysfunction and in adrenergic hyperactivity, respectively. The relationship between extreme variations in circulating catecholamines and beta-adrenergic receptor activity was studied in two patients with severe orthostatic hypotension (Shy-Drager syndrome) and in a patient with pheochromocytoma with excessive spontaneous catecholamine increases using the lymphocyte beta 2-adrenoceptor assay. In both patients with Shy-Drager syndrome, basal plasma concentrations of epinephrine and dopamine were low under resting conditions and could not be stimulated in the upright position. Norepinephrine was low in the first patient, and could not be stimulated; whereas the second patient had a normal basal concentration of norepinephrine, which could be moderately stimulated. There was no beta adrenoceptor abnormality in the first patient: however, in the second patient, there were no measurable beta-adrenoceptors on membrane fractions, whereas a population of receptors only in the low affinity state could be identified on intact cells. Alpha-adrenoceptor density on thrombocyte membranes was slightly increased in both patients with Shy-Drager syndrome and showed no substantial change during upright posture. Catecholamine increases in the pheochromocytoma patient were accompanied by a rise in blood pressure, bradycardia, and an acute up-regulation of beta-adrenoceptors. Plasma concentrations of cAMP paralleled the increase in receptor density and blood pressure. The findings in pheochromocytoma add support to the theory that an acute catecholamine stimulation gives rise to an acute beta-adrenergic sensitization leading to blood pressure elevation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559243 TI - [Circadian rhythm of the pituitary-adrenal system in health and disease]. PMID- 2559244 TI - [Difficulties in the diagnosis of primary cancer of the liver]. AB - Case records have been analyzed for 92 patients with primary cancer of the liver. The discrepancies between the clinical and postmortem diagnosis reached 31%. It appeared that the exact diagnosis depended on valid evaluation of the disease history, clinical signs and findings at special examinations rather than on the number of investigations. Associated liver cirrhosis recorded in 24% of cases put another obstacle for a precise vital diagnosis. The study has furnished evidence for a judgment on the incidence of hepatic cancer, concomitant cirrhosis, possible etiologic factors, the state of clinical diagnosis and causes of death. PMID- 2559242 TI - Comparison of glycemic response and insulin requirements after mixed meals of equal carbohydrate content in healthy, type-1, and type-2 diabetic man. AB - The postprandial insulin requirements after three mixed meals of equal carbohydrate and energy content were assessed in 10 type-1 and 12 type-2 diabetics by a glucose-controlled insulin infusion system. These were compared with the glycemic response to the same meals of 10 healthy individuals (glycemic index). In type-1 diabetics, we found the highest insulin requirements after consumption of a continental breakfast (low fibre, low protein, high fat). Ten percent less insulin was infused after milk (low fat, high protein) and 30% less after an English breakfast (high fibre, high protein). Type-2 diabetics showed no significant differences in insulin requirements between the three test meals. The glycemic response in healthy individuals had no relation to these insulin requirements. Continental and English breakfast had a similar glycemic effect, whereas milk produced only 30% of the blood glucose response observed after the continental breakfast. These results indicate that neither the carbohydrate content (exchange lists) nor the glycemic index enable prediction of postprandial insulin requirements in insulin-deficient diabetes. For this purpose, we propose the insulin-need index, elaborated by testing whole meals in closed-loop experiments with type-1 diabetics. PMID- 2559245 TI - Cytochrome c oxidase deficiency in subacute necrotizing encephalopathy (Leigh syndrome). AB - Tissues and cultured fibroblasts from two patients with Leigh syndrome (subacute necrotizing encephalopathy) were examined. A systemic defect in cytochrome oxidase was identified by enzyme assay and estimation of cytochrome concentrations. Immunochemical analysis showed a reduction of most subunits of the cytochrome oxidase complex. The rate of synthesis of cytochrome oxidase subunits, determined by labelling experiments in cultured fibroblasts, was the same in the patients and normal controls. The reduced cytochrome oxidase content of the patients' tissues must therefore result from abnormal turnover of the protein subunits. PMID- 2559246 TI - Deficiency of pyrimidine 5'-nucleotidase in human leukocytes. AB - Both erythrocytes and leukocytes from a patient with erythrocyte pyrimidine 5' nucleotidase (P5N) deficiency were shown to contain increased amounts of pyrimidine nucleotides. These findings suggested that the leukocytes were also deficient for P5N. Measurement of the P5N activity in lysates from lymphocytes or granulocytes, in the presence of inhibitors for non-specific 5'-nucleotidase or alkaline phosphatase, indeed showed a deficiency for P5N in lymphocytes and granulocytes of the patient with erythrocyte P5N deficiency. However, the P5N deficiency in the leukocytes did not cause clinical disturbances in addition to the weak haemolytic anaemia. PMID- 2559247 TI - Defective pattern of mitochondrial respiratory enzymes in mitochondrial myopathy. PMID- 2559248 TI - Estrone sulfate and sulfatase activity in human breast cancer and endometrial cancer. AB - Estrone sulfate (E1-S) in the serum and tissues of patients with breast cancer or endometrial cancer was measured by a direct radioimmunoassay without hydrolysis. The concentration of E1-S in breast cancer tissue was 1.64 +/- 0.28 ng/g wet wt (+/- SE), lower than in surrounding normal breast tissue (4.46 +/- 1.23). Estradiol-17 beta(E2)/E1-S was higher in endometrial cancer tissue than normal endometrial tissue. Estrone sulfatase activity in breast cancer tissue was 0.81 +/- 0.23 nmol/h/mg protein, higher than in surrounding normal breast tissue (0.35 +/- 0.11). These results suggest that E1-S, which is abundant in the peripheral circulation, is hydrolyzed by sulfatase in breast cancer tissue or endometrial cancer tissue and liberates free estrogens, which may stimulate the growth of these malignant tumors. PMID- 2559250 TI - Effect of vitamin D3 administration on serum 25-hydroxyvitamin D3, 1,25 dihydroxyvitamin D3 and osteocalcin in vitamin D-deficient elderly people. AB - In elderly institutionalized people, confined to bedroom and receiving no vitamin D supplementation, the frequency of vitamin D deficiency is found very high. Systematic administration of vitamin D has, therefore, been proposed to correct vitamin D deficiency. Within this context, we studied 40 elderly institutionalized subjects (mean age 80.5 + 7.2 yr) with low 25(OH)D3 concentrations (4.4 + 1.8 micrograms/l). Sixteen of them (Group I) had low serum calcium concentrations (less than 2.3 mmol/l) and 24 (Group II) had normal serum calcium concentrations (from 2.3 to 2.6 mmol/l). As hypocalcemia has been shown to regulate 1,25(OH)D3 production independent of PTH in animals and in humans, we compared their respective responses to the administration of vitamin D3. Subjects received a total dose of 15 mg (600,000 IU) of vitamin D3 divided into 3 i.m. injections at one month intervals and were explored before therapy and one and 6 months after the last dose of vitamin D3. The treatment induced a similar marked rise in 25(OH)D3 levels (from 4.1 + 1.7 to 24.4 + 8.7 micrograms/l for group I and from 5.1 + 1.8 to 27.2 + 8.0 micrograms/l for group II) in both groups but increased the 1,25(OH)2D3 concentrations only in group I (from 22.9 + 6.9 to 32.6 + 11.3 ng/l). Meanwhile serum calcium concentrations rose in group I (to low normal range i.e. 2.31 + 0.07 mmol/l) and were unaffected in group II. These results suggest that hypocalcemia is a potent stimulator of renal 1-hydroxylase in elderly people. Furthermore, a transient significant (P less than 0.01) increase in serum osteocalcin (from 10.6 + 4.1 to 14.1 + 5.9 micrograms/l) could be observed in group I which demonstrates for the first time that the osteocalcin response of osteoblasts to stimulation by 1,25(OH)2D3 is retained in very old people. PMID- 2559249 TI - Proopiomelanocortin-derived peptides, phosphoinositides, cAMP, and aldosterone secretion. AB - Since the intracellular messengers of various proopiomelanocortin-derived peptides remain ambiguous at best, we have investigated the possible involvement of phosphoinositide metabolism in aldosterone secretion evoked by alpha-MSH, beta LPH, as well as ACTH in rat and calf adrenal glomerulosa cells. We have also examined the cAMP responses in the adrenal glomerulosa cells to alpha-MSH comparing it with those of ACTH. Our results showed that neither alpha-MSH, beta LPH, nor ACTH increased inositol triphosphate (IP3) or other inositol phosphates in adrenal glomerulosa cells while increasing aldosterone secretion from the same cells. Angiotensin II, known to cause hydrolysis of the phosphoinositides, increased IP3 in these adrenal cells in a dose-dependent manner. Both ACTH and alpha-MSH raised the cAMP levels in the calf adrenal glomerulosa cells, although the magnitude of the increase of cAMP in response to ACTH was greater. These findings suggest that IP3 as a mediator of alpha-MSH- and beta-LPH-induced aldosterone secretion is not likely and other mediator(s) may be involved. PMID- 2559251 TI - The dynamics of progesterone output in perifused granulosa layer of the Japanese quail (Coturnix coturnix japonica): response to adenosine 3',5'-cyclic monophosphate and aminoglutethimide. AB - A perifusion system has been developed in which the rates of progesterone output of quail granulosa layer in response to steroidogenic stimuli, can be measured under dynamic condition. In response to a continuous perifusion for 8 h with dibutyryl cyclic AMP (dbcAMP), there was a gradual increase in the rate of progesterone output, which reached a plateau at 4-5 h of perifusion. When progesterone output was inhibited by aminoglutethimide, a reversible inhibitor of the cholesterol side chain cleavage reaction, dbcAMP caused the accumulation of cholesterol in mitochondria. The rate of progesterone output after the release of inhibition was much faster when the tissue had been stimulated with dbcAMP during the inhibition. This indicates that dbcAMP stimulates progesterone production by increasing the steroidogenic cholesterol in mitochondria. However, the resumption of progesterone output after the release of inhibition was not observed unless the tissue was again stimulated with dbcAMP. This indicates that the mitochondrial cholesterol is not readily available for the cholesterol side chain cleavage enzyme, and there is another step of stimulation of dbcAMP for progesterone output of quail granulosa layer. PMID- 2559252 TI - Inhibition of 17 alpha-hydroxylase/C17-C20 lyase by bifluranol and its analogues. AB - A simple assay for the measurement of the activities of both 17 alpha-hydroxylase and C17-C20 lyase is described. No extraction procedures are required. The separation of substrate and products is achieved using HPLC which allows the collection of the components of interest and the monitoring of the recovery of various steroids. Using this assay, bifluranol (known to show anti-prostatic activity in vivo) and some analogues were tested for inhibitory activity towards these enzyme activities. Each compound was active, although less potent than ketoconazole, and this activity may contribute towards the in vivo action. PMID- 2559253 TI - Studies on spirolactone steroid antagonists in ACTH-induced hypertension in sheep. AB - This study investigated the anti-mineralocorticoid potency and haemodynamic effects of a series of mineralocorticoid antagonists of the spirolactone type (RU 28318, spironolactone, K-prorenoate, K-canrenoate and canrenone), for their ability to prevent the development of ACTH-induced hypertension in conscious sheep. In vivo bioassay, using aldosterone dependent changes in parotid salivary [Na+]/[K+] of sodium depleted adrenalectomized sheep, showed spironolactone was the most potent anti-mineralocorticoid tested. Infusions of the antagonists at equal doses alone for 4 days demonstrated that none affected mean arterial pressure, except for K-prorenoate which exhibited slight pressor activity. All the antagonists produced a natriuresis. Some of the steroid antagonists of the spirolactone group blocked the development of ACTH hypertension in sheep, spironolactone being the most effective. This study provides additional evidence for an essential mineralocorticoid component in ACTH-induced hypertension. PMID- 2559254 TI - Effects of ACTH on the last step of aldosterone biosynthesis. AB - The production of tritiated aldosterone and tritiated SM (a saponifiable 18 hydroxycorticosterone derivative) by rat adrenals were studied at various incubation times in absence or presence of two concentrations of ACTH. Tritiated 18-hydroxycorticosterone or 18-deoxyaldosterone served as precursors. The lower ACTH concentration (150 pM) increased the production of tritiated aldosterone. Whereas, the higher ACTH concentration (1.5 microM) stimulated tritiated aldosterone production at shorter incubation time (30 min), while after 60 min it inhibited. This time dependency would reflect variations in the levels of endogenous steroids. On the other hand, the effects of ACTH on tritiated SM production were opposite to those on tritiated aldosterone. In effect, while 150 pM ACTH inhibited SM production, 1.5 microM ACTH stimulated it. These results suggest that ACTH promotes opposite effects on the productions of aldosterone and SM and therefore both productions would be coordinated under the regulation of ACTH. PMID- 2559255 TI - A simple method for producing graded aortic insufficiencies in rats and subsequent development of cardiac hypertrophy. AB - Selective perforation of the right cusp of the aortic valve alone (1V) or in combination with that of the left valve cusp (2V) by a plastic rod inserted from the right common carotid artery was performed to cause aortic insufficiency (AI) in rats. The development of cardiac hypertrophies subsequent to these interventions to cause graded cardiac volume overloads was followed up every week till the end of week 4 after the operations by radiographic measurement of the cardiothoracic ratio (CTR). Rats were killed 2 and 4 weeks after the operations, and the density of beta-adrenoceptors in hypertrophied cardiac muscle membranes was determined by radioligand binding assay and compared with sham-operated (sham) rats. Production of AI resulted in an enlargement of the cardiac shadow in 1VAI and 2VAI rats, with significant graded increases in the CTR at the end of week 3. The occurrence of graded cardiac hypertrophies was well revealed in graded increases of the heart weight/body weight ratio among the sham, 1VAI, and 2VAI rats at the end of both week 2 and week 4. There was a tendency to stepwise increases due to graded AI in the density of binding sites for 3H dihydroalprenolol, with a significant increase in 2VAI rats at the end of week 2. At the end of week 4, plasma norepinephrine levels of 2VAI rats was raised significantly as compared with the sham rats. Selective perforation of either one or two cusps of the aortic valve can be performed with ease using touch alone and provides us with a simple method for producing graded cardiac hypertrophies in rats due to pure volume overload. PMID- 2559256 TI - Mechanistic clues to the mutagenicity of alkylated DNA bases: a theoretical study. AB - Experiment indicates that the N7-guanine site in DNA is not "promutagenic" (mutation-inducing) on alkylation, while the O6-guanine and O4-thymine sites are so. These differences in nucleic acid template activity are attributed to corresponding differences in acidity of the Watson-Crick hydrogen bonding protons. Mechanistic indicators for ease of Watson-Crick proton loss are calculated using molecular orbital theory for DNA bases alkylated at the N7 guanine, O6-guanine and O4-thymine sites. Their values point to a definite favouring of the proton loss for the O-alkylated bases compared to the N7 alkylguanines. This may suggest the possibility that, at biological pH, the O alkylated bases deprotonate readily while the N7-alkylguanines do not, thus accounting for observed differences in promutagenicity and nucleic acid template activity. PMID- 2559257 TI - A model for the initiation of replication in Escherichia coli. AB - The role of the protein DnaA as the principal control of replication initiation is investigated by a mathematical model. Data showing that DnaA is growth rate regulated suggest that its concentration alone is not the only factor determining the timing of initiation. A mathematical model with stochastic and deterministic components is constructed from known experimental evidence and subdivides the total pool of DnaA protein into four forms. The active form, DnaA.ATP, can be bound to the origin of replication, oriC, where it is assumed that a critical level of these bound molecules is needed to initiate replication. The active form can also exist in a reserve pool bound to the chromosome or a free pool in the cytoplasm. Finally, a large inactive pool of DnaA protein completes the state variables and provides an explanation for how the DnaA.ATP form could be the principal controlling element in the timing of initiation. The fact that DnaA protein is an autorepressor is used to derive its synthesis rate. The model studies a single exponentially growing cell through a series of cell divisions. Computer simulations are performed, and the results compare favorably to data for different cell cycle times. The model shows synchrony of initiation events in agreement with experimental results. PMID- 2559258 TI - Kinetic approach with ab initio MO method on ionic selectivity and size in sodium channel. AB - Three kinds of models for ionic selectivity and size of the filter in sodium channel have been treated by using ab initio molecular orbital (MO) calculations with MINI-3 and MIDI-3* basis sets. A three-components system, HCO2M-H2O (M = Li+, Na+ or K+), is acceptable for describing experimental facts well. Thermochemical parameters obtained from harmonic vibrational analysis with MINI-3 basis sets, for the translocation of the permeant metal cations in the HCO2M-H2O system, are that the activation enthalpies for Li+, Na+ and K+ are 7.0, 6.4 and 23.4 kJ/mol, and also the free energies of activation are 10.6, 1.5 and 19.0 kJ/mol, respectively. These results are qualitatively in good correspondence with experimental facts of the ion selectivity of the channel. One of water molecule was found to have a key role in the translocation of the permeant cations. PMID- 2559259 TI - Analysis of cellular heterogeneity in the response of human leukemic cells to photosensitization induced by pyrene-containing fatty acid. AB - Incubation of cells with 12-(1-pyrene) dodecanoic acid (P12) followed by irradiation with ultraviolet light at 366 nm (UVA) resulted in cytotoxicity. We compared the photosensitivity to UVA irradiation of various human myelo-monocytic leukemic cell lines, their intra- and inter-clonal variability and correlated their photosensitivity to P12-uptake and metabolism. The fluorescence properties of pyrene were utilized for flow cytometric analysis of cell distribution with respect to P12-uptake as well as for sorting subpopulations differing in their fluorescence. Spectrofluorometric analysis of the total cell-associated fluorescence and of the cellular lipids-associated fluorescence were also carried out. Considerable heterogeneity in P12-uptake and photosensitivity was found not only among cell lines, but also in the response of different clones and among the individual cells in specific clonal populations. Within a clone, photosensitivity was related to the amount of P12 taken up by the individual cells, while among different cell lines and their clones the photosensitivity was correlated with the proportion of cellular pyrene-linked phospholipids. The larger the fraction of pyrene-linked phospholipids within the cell--the more sensitive it was to UVA irradiation. Photosensitivity could be affected by changing the proportion of cellular pyrene-linked phospholipids. Cells treated with cAMP showed an increase in total P12-uptake, but the proportion of pyrene-linked phospholipids was reduced, resulting in lower photosensitivity. These findings, demonstrating that by manipulating lipid metabolism photosensitivity can be modified, may prove useful in a clinical setting for selective photosensitization of malignant cells. PMID- 2559260 TI - Recent development in calcium channel antagonists. AB - The introduction of the Ca2+ channel antagonists, including the clinically available verapamil, nifedipine and diltiazem, into cardiovascular medicine served to initiate much work directed to the elucidation of their mechanisms of action at voltage-dependent Ca2+ channels. The Ca2+ channel ligands (both activator and antagonist) interact with the channel in a state-dependent manner associating preferentially with open states (activators) or open and inactivated states (antagonists). Both frequency- and voltage-dependent interactions occur with activator and antagonist drugs. These effects underlie the antiarrhythmic activity of verapamil and the vascular smooth muscle selectivity of 1,4 dihydropyridines. Selectivity of action occurs from a combination of factors including the relative Ca2+ demands of the system, the stimulus mode and state dependence of interaction, the agonist/antagonist character of the channel ligand and the category of Ca2+ channel involved. PMID- 2559261 TI - Carboplatin in association with etoposide and either adriamycin or epirubicin for untreated small cell lung cancer: a dose escalation study of carboplatin. UCL Clinical Oncology Group. AB - A multi-center, open trial was conducted to determine the maximal tolerable dose of carboplatin in combination with conventional doses of both etoposide and an anthracycline for the treatment of previously untreated small cell lung cancer (SCLC) patients. Ninety-five patients [48 with limited disease (LD) and 47 with extensive disease (ED)] received a total of 376 courses of treatment. Carboplatin was given on day 1 at a dose of 250 mg m-2 in 60 courses, 300 mg m-2 in 69, 330 mg m-2 in 236 and 350 mg m-2 in 11, with 120 mg m-2 etoposide on days 1, 3 and 5 and either 40 mg m-2 adriamycin or 60 mg m-2 epirubicin on day 1. Epirubicin was not administered before carboplatin reached the dose of 330 mg m-2. Courses were repeated every 3 weeks. The main toxicity was hematological. The first course of therapy induced a dose-dependent decrease of leucocyte, neutrophil and platelet counts: all patients, except one, who received 350 mg m-2 carboplatin had a neutropenia below 200 microliters-1 and a thrombopenia below 100,000 microliters 1. Three patients died of septicemia. Other toxicities were well tolerated. After three courses, patients were re-staged by performing a mandatory fiberoptic bronchoscopy and a thoracic computed axial tomography (CAT). The overall objective response rate for 86 evaluable patients was 91% (98% for LD) with 21% complete remissions (30% for LD). All 23 hepatic and six brain sites, evaluable after chemotherapy alone, responded.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559262 TI - Treatment of small cell lung cancer with induction chemotherapy followed by late intensification. AB - Seventy-seven patients with small cell lung cancer were entered on a protocol comprising induction chemotherapy with cyclophosphamide 1 g m-2, adriamycin 40 mg m-2 and vincristine 1.4 mg m-2 (CAV) every 21 days for four to six cycles. The overall response rate was 72.8%. Twenty-six patients achieving complete remission received intensification with two further cycles of ifosfamide 5 g m-2, mesna 8 g m-2, methotrexate 30 mg m-2 and etoposide 100 mg m-2 per day for 3 days (IME). Six of the 15 patients in partial remission following CAV achieved a further remission on IME (response rate 40%). Median survival in the limited disease group was 11 months compared with 7 months in the extensive disease patients and four patients are alive at more than 2 yr follow up. There was no significant prolongation of the median survival (11 months) seen in those patients in complete remission who had negative second bronchoscopy examination. This sequential six drug regime produces high response rates in small cell lung cancer, and there is evidence of lack of cross-resistance between CAV and IME. PMID- 2559263 TI - Accidental overdose of mitoxantrone in three patients. AB - Mitoxantrone is a new effective antineoplastic agent with activity against a wide range of tumors. Compared with the anthracycline drugs doxo- and daunorubicin, it exhibits a clearly lower toxicity and, most importantly, a reduced cardiotoxicity. The analysis of the side-effects recorded after accidental overdosage of the drug gives additional insight into its tolerability. Here we describe our observations in three patients who inadvertently received 100 mg m-2 (two pts) and 183 mg m-2 (one pt) as single slow bolus injections. The main side effects were moderate nausea and vomiting, shaking chills, and profound but reversible neutro- and thrombocytopenia. There was no immediate cardiac toxicity. One patient with extensive previous daunomycin exposure developed congestive heart failure after 4 months. Two patients were not evaluable for late cardiac complications because of early death due to tumor progression. PMID- 2559265 TI - [31-year-old man with disseminated neoplasms, chronic diarrhea syndrome and episodes of paroxysmal supraventricular tachycardia]. PMID- 2559264 TI - Treatment of small cell lung cancer by eight weeks chemotherapy. AB - Eighty-two patients with small cell lung cancer (SCLC), 32 with limited disease, were treated with alternating chemotherapy. Eight courses were administered at weekly intervals, and responding patients received radiotherapy to sites of bulk disease. Overall response rate was 76.8%. Overall median survival was 265 days, 408 days in patients with limited disease. The median symptom free period after chemotherapy was completed was 123 days. These results are comparable with those in reports of chemotherapy of longer duration and warrant the further investigation of short duration treatment in this disease. PMID- 2559267 TI - [Carcinomatous ventriculitis]. PMID- 2559266 TI - [Malignant insulinoma: lack of response to the somatostatin analog SMS 201-955]. PMID- 2559268 TI - [Bowenoid papulosis]. AB - Bowenoid papulosis is a clinical-histopathologic entity described in the last ten years. The appearance and publication of an increasing number of cases in the world literature and the advances in laboratory techniques for detection of papilloma-virus, lead to a better knowledge of its clinical picture, etiology, prognosis and therapy. Upon this basis, a therapeutic and clinical approach is suggested. PMID- 2559269 TI - [Granular cell tumor of the skin]. AB - Two adult males with cutaneous granular cell myoblastoma, localized on the leg and pubic area, are described. Atypical localizations of Abrikossoff's tumor are reviewed pointing out that the upper trunk is the most frequent of these localizations. Histopathogenesis of that tumour is also considered. PMID- 2559270 TI - [Indications and current developments in liver transplantation]. AB - Today, liver transplantation has become a routine treatment in many centres and will be increasingly applied in the future. Principal indications for liver transplantation are endstage chronic liver diseases, namely cirrhoses of various origin, fulminant hepatic failure, metabolic liver disease, and rarely non resectable malignant tumors of the liver. Clinical and biochemical parameters indicative of the indication for liver transplantation can be stated with greater confidence. Advances in organ preservation with a new cold storage solution (University of Wisconsin solution) and further refinements in surgical technique and immunosuppression with application of multimodal protocols have led to a one year-survival rate of 70%-80% in experienced centers. Since October 1988, 32 patients underwent liver transplantation in Berlin, currently, 31 patients are alive, while one patient died due to systemic fungal infection. PMID- 2559271 TI - [Endoscopy and biopsy of the pancreatic duct--a new method of pancreas diagnosis]. AB - Newly developed endoscopes make it possible to inspect the major papilla, the pancreatic and choledochal ducts without previous papillotomy. First results of this new method in surgical specimens and patients are presented. Ductal dilatations, and irregularities as well as secretion plugs and calcifications are characteristic phenomenons of chronic pancreatitis; the diagnosis of a malignant pancreatic tumour can be made by observing an irregular compression of the pancreatic duct and the infiltration of its wall. Perhaps by this method precursors of ductal pancreatic cancer will be recognized. We assume that ERCP will be supported by this method. PMID- 2559272 TI - A pilot study of diethyldithiocarbamate in patients with acquired immune deficiency syndrome (AIDS) and the AIDS-related complex. AB - We investigated the use of diethyldithiocarbamate (DTC, or Imuthiolr, Merieux Institute) as a therapeutic agent in patients with Acquired Immune Deficiency Syndrome (AIDS) and AIDS-Related Complex (ARC). Patients were prospectively stratified and randomized to receive DTC 200 mg/m2 intravenously weekly for 16 weeks or no therapy, followed by crossover to the opposite arm for an equal period. Forty-four patients were entered and forty were evaluable. There was a statistically significant decrease in symptoms in the DTC treated patients compared to the controls (p = .002). There was a significant improvement in lymphadenopathy in the treated patients compared to the controls (p = .005). One patient showed disappearance of splenomegaly, one clearing of antifungal agent resistant perianal moniliasis, and one clearing of hairy leukoplakia. No significant differences in progression were noted. No changes were seen in any of the immunological parameters measured. There was no significant toxicity. Because of the changes in symptoms and in lymphadenopathy, we suggest that further study of DTC, both alone and in combination with other agents, may be indicated. PMID- 2559273 TI - Antagonism of kappa opioid mediated effects in the rat by cyclo(Leu-Gly). AB - The effect of cyclo(Leu-Gly) on U-50,488H- induced pharmacological actions was determined in male Sprague-Dawley rats. Intraperitoneal (i.p.) administration of U-50,488H to rats produced analgesia (tail-flick) and increased urinary output. Cyclo (Leu-Gly) (1-4 mg/kg, s.c.) antagonized the analgesic response to U-50,488H (25 mg/kg; i.p.). A dose of 10 mg/kg (i.p.) of U-50,488H increased the spontaneous urinary output which was antagonized by cyclo (Leu-Gly) (1-4 mg/kg; s.c.). To determine whether cyclo (Leu-Gly) was acting as a kappa-opioid receptor antagonist, the effect of cyclo (Leu-Gly) on the binding of [3H]ethylketocyclazocine (EKC) to membranes of rat cerebral cortex and spinal cord was determined. The IC50 values of cyclo(Leu-Gly) in displacing [3H]EKC from its binding sites in cortex and spinal cord were 1.44 and 0.40 mM, respectively. Chronic administration of U-50,488H (25 mg/kg; i.p., b.i.d.) for 4 days induced tolerance to its analgesic effect. The latter was not affected by cyclo(Leu-Gly) (2 to 8 mg/kg; s.c.) given once a day for 4 days. It is concluded that cyclo(Leu Gly) antagonizes acute actions of U-50,488H and that such effects of cyclo(Leu Gly) are not mediated via a direct action on kappa-opioid receptors. PMID- 2559274 TI - Lack of effect of the opioid antagonist, naltrexone, on the in situ growth of C 1300, N1E-115 and NS206 murine neuroblastoma tumor cell lines. AB - Attempts have been made to confirm previously reported results which demonstrated that the opioid antagonist, naltrexone, altered the in situ growth of murine neuroblastoma tumors. Adult male A/J mice were injected with tumor cells from three different cell lines of murine neuroblastoma; the spontaneously arising C 1300 line, the adrenergic clonal line N1E-115, and the cholinergic clonal line NS20Y. Naltrexone was administered daily in doses of 0.1, 0.4, or 10.0 mg/kg subcutaneously, to replicate the reported experimental design. In contrast to previous studies, we were unable to demonstrate any effect of naltrexone on in situ growth or other characteristics of tumors produced by the C-1300, N1E-115 or NS20Y murine neuroblastoma cell lines. Ligand binding studies have demonstrated the presence of high levels of opiate binding sites on membranes prepared from the NS20Y clonal cell line and low levels on the membranes of the C1300 tumor line. PMID- 2559275 TI - Inhibition of oxytocin-induced but not angiotensin-induced rat uterine contractions following exposure to sodium sulfide. AB - Low concentrations (0.15-15 microM) of sodium sulfide reversibly attenuated the contractile response of the isolated rat uterus to oxytocin without affecting angiotensin II responsiveness. These findings suggest that functionally important disulfide bonds in the rat uterine oxytocin receptor, but not the angiotensin receptor, are sensitive to hydrosulfide ion. Reduction of oxytocin receptors by hydrosulfide ion may be a mechanism by which low levels of H2S delay parturition in rats. PMID- 2559276 TI - Benextramine and nifedipine distinguish between sub-classes of alpha 1 adrenoceptors. AB - The effects of benextramine and nifedipine were examined on the dose-diastolic pressure response to methoxamine in pithed normotensive rats. Benextramine (3, 6 and 12 mg/Kg) displaced the dose-response curve to methoxamine to the right. Maximum response was reduced after the administration of 12 mg/Kg benextramine. Nifedipine (0.1 and 0.3 mg/Kg) also caused the dose-response curve to methoxamine to be displaced to the right with reduction in maximum response. Nifedipine effects were additive with an increase in the EC50 values as well as reduction in the maximum response after pretreatment with benextramine (3 and 6 mg/Kg). However, at the highest dose of benextramine the effects of nifedipine were diminished and no longer apparent. It is concluded that benextramine may have alkylated a nifedipine sensitive site on the alpha 1-adrenoceptors. PMID- 2559277 TI - Effect of methotrexate on the intestinal mucosa of PGE2-treated rats. AB - In the present study, we aimed to protect the intestinal mucosa from small bowel damage in methotrexate (MTX)-treated rats. The protective effect of prostaglandin E2 (PGE2) was investigated. Ileal integrity was evaluated making use of different biochemical parameters: content of sucrase and maltase activities, contents of DNA, proteins, AMPc, PGE2, putrescine (Put), spermine (Spm) and spermidine (Spd). Rats were orally administered 0.5 ml of NaCl solution (0.9%) containing or not containing 400 micrograms.ml-1 of PGE2 twice daily, during three or ten days. Half an hour after the 18th ingestion of PGE2, 0.5 ml of NaCl solution (0.9%) containing MTX (16 mg.ml-1) was injected intravenously. Rats were killed exactly 48 hours after this injection. MTX had no effect on the Put content, increased the AMPc content and decreased the contents of DNA, proteins, Spm, Spd, PGE2 and sucrase or maltase activity. PGE2 had no effect on the biochemical parameters we studied, except on the contents of DNA (10-day treatment) and of PGE2 (3- and 10 day treatment). When MTX was injected after PGE2 treatment, as compared with what was observed when MTX was used as reported above, we observed--an increase in spermine content after 3-day PGE2 treatment and- an increase in the contents of DNA, Spm, Spd and disaccharidase activity after 10-day PGE2 treatment. No other significant variation in the other biochemical parameters was recorded, whatever the duration of the PGE2 treatment. These results indicate that PGE2 could partially protect the intestinal mucosa against the biochemical effects of MTX. Other experimental conditions may need to be chosen in order to obtain a better cytoprotective effect of PGE. PMID- 2559278 TI - Detergent induced changes in serum lipid composition in rats. AB - The influence of in vivo administration of detergents on serum lipid composition was studied in rats. Male Wistar rats received 50 mg Emulgen 913 (polyoxyethylene nonylphenylether, a nonionic detergent) or SDS (sodium dodecylsulfate, an anionic detergent) per kg of body weight intraperitoneally for 3 consecutive days. Emulgen 913 and SDS administration increased the level of cholesterol esters and phospholipids, respectively. But Emulgen 913 administration reduced the level of triglycerides in the serum, and SDS administration reduced also the levels of triglycerides and cholesterol esters. In spite of the changes in serum lipid composition, the administration of these detergents did not affect the amount of total lipids in rat serum. The proportion of palmitic, oleic, and docosahexaenoic acids in phospholipids was decreased by the administration of Emulgen 913 while the level of arachidonic acid was raised. However, the level SDS administration had no effect on the fatty acid composition of the serum phospholipids. On the other hand, both Emulgen 913 and SDS administration showed an effect on the fatty acid composition of triglycerides. It is postulated that liver damage due to administration of detergents is responsible for the changes in serum lipid and fatty acid composition in detergent-treated rats. PMID- 2559279 TI - Inhibitory effect of stearidonic acid (18:4 n-3) on platelet aggregation and arachidonate oxygenation. AB - The effect of stearidonic acid (18:4 n-3) present in fish and some plant oils, such as black currant seed oil, was studied on human platelets. When added to platelets simultaneously with collagen, arachidonic acid or endoperoxide mimetic U46619, 18:4 n-3 appeared as a weak inhibitor of platelet aggregation. In addition, 18:4 n-3 did not alter the metabolism of exogenous arachidonic acid. In contrast, when preincubated with platelets after precoating onto albumin, 18:4 n 3 inhibited platelet aggregation induced by thrombin, collagen, arachidonic acid or U46619, and was as potent as eicosapentaenoic acid (20:5 n-3) tested under similar conditions. Stearidonic acid also altered the endogenous arachidonate oxygenation stimulated by low doses of thrombin, but to a significantly lesser extent than did 20:5 n-3. It seems therefore that, in addition to competing with endogenous arachidonate metabolism, 18:4n-3 may affect platelet aggregation by another mechanism. PMID- 2559280 TI - Apparent lack of effect of obesity on the soluble phosphatidic acid phosphatase activity in human adipose tissue. AB - In view of previous reports that the activity of the Mg(++)-dependent phosphatidic acid phosphatase in adipose tissues of rat and mouse is elevated in obesity, we attempted to assay this activity in biopsies of human omental adipose tissue obtained from normal-weight and morbidly obese subjects in connection with operations. The major portion of the phosphatidic acid phosphatase activity was found in the cytosol, and the small amount found in the microsomal fraction was too low for accurate measurement. It was not possible to assay the activity in the crude cytosol. After precipitation with ammonium sulfate, however, the enzyme activity was linear with both the incubation time and the concentration of enzyme. It was not possible to obtain substrate saturation of the enzyme under the conditions employed. When assayed in the presence of a high concentration of substrate (0.6 mmol/l) the activity obtained in normal-weight patients, 7.8 +/- 2.4 nmol/mg protein/min (n = 10), was not significantly different from that in morbidly obese patients, 5.6 +/- 0.8 nmol/mg protein/min (n = 10). There was no relation between the size of adipose cells and phosphatidic acid phosphatase activity. Furthermore, there was no apparent relation between phosphatidic acid phosphatase activity in omental adipose tissue and that in the liver. The findings suggest that the increased biosynthesis of triglycerides in human obesity is not associated with an increased capacity of the soluble phosphatidic acid phosphatase in adipose tissue. PMID- 2559283 TI - Pentobarbital suppresses human brain sodium channels. AB - Ion channels, key components in neuronal signal transmission and processing, are likely to be important molecular sites of anesthetic action. Sodium channels from human brain tissue were incorporated into planar lipid bilayers in the presence of batrachotoxin and exposed to the anesthetic pentobarbital. This barbiturate, in a dose-dependent manner and at clinically relevant concentrations, reduced fractional channel open time independent of membrane potential, and interfered with the steady-state activation process. PMID- 2559281 TI - Guinea pig epidermis generates putative anti-inflammatory metabolites from fish oil polyunsaturated fatty acids. AB - Clinical studies have indicated that dietary fish oil may have therapeutic value in the treatment of psoriasis, a hyperproliferative, inflammatory skin disorder characterized by elevated LTB4. To evolve a possible mechanism for these beneficial effects, we determined the metabolic fate of fish oil derived n-3 fatty acids in the skin. Specifically, we incubated guinea pig epidermal enzyme preparations with [3H]eicosapentaenoic acid (20:5 n-3) and [14C]docosahexaenoic acid (22:6 n-3). Analyses of the radiometabolites revealed the transformation of these n-3 fatty acids into n-6 lipoxygenase (arachidonate 15-lipoxygenase) products: 15-hydroxyeicosapentaenoic acid (15-HEPE) and 17-hydroxydocosahexaenoic acid (17-HDHE), respectively. Since 15-lipoxygenase products have been suggested as possible endogenous inhibitors of 5-lipoxygenase (an enzyme which catalyzes the formation of LTB4) we tested the ability of 15-HEPE and 17-HDHE in vitro to inhibit the activity of the 5-lipoxygenase. Incubations of these metabolites with enzyme preparations from rat basophilic leukemia (RBL-1) cells demonstrated that 15-HEPE (IC50 = 28 microM) and 17-HDHE (IC50 = 25 microM) are respectively potent inhibitors of RBL-I-5-lipoxygenase. The inhibitory potential of these fish oil metabolites provides a possible mechanism by which fish oil might act to decrease local cutaneous levels of LTB4, and thereby alleviate psoriatic symptoms. PMID- 2559282 TI - Detection of phosphatase production by Staphylococcus species: a new method. AB - The ability of staphylococci to produce phosphatase is an important character in the identification of species in the genus. Current methods are slow and difficult to interpret. A test is described which uses p-nitrophenyl phosphate as the substrate. It was evaluated by testing 1035 reference and clinical strains, and also compared with an alternative method. All strains of Staphylococcus aureus and 83% of Staphylococcus epidermidis were phosphatase positive: all strains of Staphylococcus saprophyticus were negative. The method proved to be an accurate, reliable, economic and relatively rapid technique to detect phosphatase production. PMID- 2559284 TI - [Cardiac rhythm disorder in infection caused by cytomegalovirus in patients with kidney transplant]. AB - A higher incidence of arrhythmias was observed in 16 renal allograft recipients infected with Cytomegalovirus (CMV), as compared to 26 patients who were transplanted during the same period and did not contract the infection (43.7% vs. 3.8%; p less than 0.01). There was a highly significant difference in the incidence of sinus tachycardia (37.5% infected vs. 0% not infected; p less than 0.005), while the incidences of atrial fibrillation and ventricular ectopic beats were not significantly different. The arrhythmias always appeared in the early phases of the infection and disappeared after a certain period, lasting from one week to months. It is proposed that arrhythmias arising during CMV infection may be due to a myocardial CMV involvement. PMID- 2559285 TI - Erythrocyte Na+/K+ ATPase activity measured with 23Na NMR. AB - A 23Na NMR assay for measurement of erythrocyte Na+/K+ ATPase activity is presented. Using the nonpermeant shift reagent dysprosium tripolyphosphate the signals of intra- and extracellular sodium are separated, enabling measurement of sodium fluxes nondestructively, without the need to physically separate the cells from their environment. By increasing membrane permeability with nystatin we have shown that the assay allows the detection of differences in membrane permeability. With low doses of nystatin the ouabain-sensitive sodium flux increased more than twofold. With high doses of nystatin the Na+/K+ pump could not prevent an almost total equilibration of intra- and extracellular sodium. All sodium that entered the cells remained NMR visible, proving that sodium influx can be measured quantitatively. 31P NMR spectra taken before and after the assay revealed a slight acidification of the cells and no significant change in ATP concentration. No evidence of Dy3+ entering the cell was observed. PMID- 2559286 TI - Intrauterine fetal brain NMR spectroscopy: 1H and 31P studies in rats. AB - Fetal brain metabolism was investigated in utero noninvasively using multinuclear nuclear magnetic resonance spectroscopy in rats at two representative prenatal stages: early (17-18 days) and late (20-21 days) stages. Phosphorus-31 (31P) spectroscopy revealed that phosphocreatine is significantly lower in the early stage and increases to the level of early neonates by the late prenatal stage. Intracellular pH at the early stage was found to be strikingly high (7.52 +/- 0.21) and decreased to a level similar to that of neonates by the late stage (7.29 +/- 0.07). Phosphomonoester levels at both stages were similar to the values reported for early neonates. Water-suppressed proton (1H) spectroscopy demonstrated a distinctive in vivo fetal brain spectral pattern characterized by low levels of N-acetyl aspartate and high levels of taurine. High-resolution proton spectroscopy and homonuclear chemical-shift correlate spectroscopy of brain perchloric acid extracts confirmed these in vivo findings. In vitro 31P spectroscopy of acidified chloroform methanol extracts showed the characteristic membrane phospholipid profiles of fetal brain. The phosphatidylethanolamine (PE) to-phosphatidylcholine (PC) ratio (PE/PC) did not show significant changes between the two stages at 0.40 +/- 0.11, a value similar to that of early neonates. PMID- 2559288 TI - Correction of distortions due to the pulsed magnetic field gradient-induced shift in B0 field by postprocessing. AB - Gradient pulses used in most localization techniques induce time-dependent shifts in B0 that strongly distort NMR signals. A postprocessing correction method requiring no additional hardware is proposed. The phase shift produced by the B0 shift is calculated from a reference free induction decay (FID) and used to correct any FID acquired with the same sequence. PMID- 2559287 TI - In vivo 31P and 2H NMR [corrected] studies of rat brain tumor pH and blood flow during acute hyperglycemia: differential effects between subcutaneous and intracerebral locations. AB - Surface coil NMR spectroscopy was used to monitor the hyperglycemia-induced alterations in pH and blood flow in vivo in C6 gliomas implanted both subcutaneously and intracerebrally in rats. Tumor pH was calculated from the chemical shift difference between PCr and Pi in the 31P NMR spectra. Subcutaneous glioma pH decreased 0.8 units by 1 h after intraperitoneal administration of an aqueous 50% glucose solution (6 g glucose per kg body weight). In contrast, hyperglycemia failed to significantly alter the pH of intracerebral gliomas which were monitored for 90 min following administration of glucose. Tumor blood flow (TBF) was determined both pre- and post-glucose administration using deuterium NMR by monitoring the time course of D2O washout following intratumoral injection of saline D2O. Subcutaneous and intracerebral TBF were found to have an average change of -78.1% (range -47.4 to -93.3%, n = 5) and -21.1% (range +6.0 to -37.8%, n = 9), respectively. In addition, laser Doppler blood flow measurements of rat skin and subcutaneous glioma revealed a dramatic reduction in blood flow in both tissues following glucose administration. These results indicate that the effects of acute hyperglycemia are site dependent and that hyperglycemia alone is not beneficial for inducing intracellular acidosis in intracerebral tumors. PMID- 2559289 TI - In vivo human localized 1H spectroscopy. PMID- 2559290 TI - In vivo localized proton spectroscopic studies of human gastrocnemius muscle. PMID- 2559292 TI - Phospholipid bilayer contribution to 31P NMR spectra in vivo. AB - The magnetic field-dependent phosphodiester (PDE) signal found in 31P NMR spectra of liver and brain has been studied using saturation transfer and proton decoupling techniques. This PDE component, which accounts for as much as 45% of the signal in vivo, has been identified as primarily phospholipid bilayer with a small contribution from a motionally averaged macromolecule(s). PMID- 2559291 TI - Studies of the metabolism of human breast cancer spheroids by NMR. AB - Spheroids present a model for tumor behavior in vivo. In NMR studies, a problem of spheroid adhesion occurs. Here, we present a method which overcomes this problem and show the variations in phosphate metabolite concentrations as well as the rate of glucose metabolism as a function of spheroid age. PMID- 2559294 TI - Independent insertion of Alu elements in the human ribosomal spacer and their concerted evolution. AB - A 2,700-bp segment of human ribosomal DNA (rDNA) spacer upstream of the rRNA promoter contains a set of four Alu elements, two in the direction of rRNA transcription and two in the opposite orientation. We report and compare the sequences of these Alu elements found in three rDNA clones and seek to determine the origin of the cluster, either from a single insertion followed by duplications or from multiple simultaneous or independent insertions. The high (20%-27%) divergence among members of a set and the lack of similarity/complementarity of sequences flanking different members of the set demonstrate the independent insertion of each of the four Alu elements into A rich sequences on the appropriate strand of the rDNA. We also demonstrate that the Alu sets found in different rDNA repeats are subject to concerted evolution, yielding divergences of only 0.4%-3% between Alu elements in equivalent positions. However, the pairs of adjacent similarly oriented Alu elements do not show reduced divergence, indicating that there is no recombination or gene conversion between similarly oriented but not equivalently positioned Alu elements. Finally, crossing-over must occur in the rDNA junction region between Alu element 3 and the nonribosomal sequences at the telomere end of the acrocentric chromosome, so that the Alu elements of the terminal rDNA repeats and the terminal repeats themselves evolve in concert with the rDNA repeats located internally in the tandem array. PMID- 2559293 TI - The CO-I and CO-II region of honeybee mitochondrial DNA: evidence for variation in insect mitochondrial evolutionary rates. AB - The sequence of a region of honeybee (Apis mellifera ligustica) mitochondrial DNA, which contains the genes for cytochrome c oxidase subunits I and II (CO-I and CO-II) and inferred genes for tRNA(Asp), tRNA(Leu)UUR, tRNA(Lys), and tRNA(Trp), is presented. The region includes the segment previously identified as incurring a length increase in some other bee strains, including Africanized bees. The sequence information of this study and of that by Vlasak et al. shows that several shifts of tRNA genes have occurred between Apis and Drosophila, but shifts of other kinds of genes have yet to be demonstrated. The CO-I and CO-II gene sequences are both more A+T rich than are the corresponding Drosophila genes. Parsimony analyses using the mouse and Xenopus sequences as outgroups show significantly more amino acid substitutions on the branch to Apis (120) than on that to Drosophila (44), indicating a difference in the long-term evolutionary rates of hymenopteran and dipteran mtDNA. PMID- 2559295 TI - Exonuclease activity that degrades histone mRNA is stable when DNA or protein synthesis is inhibited. AB - The induction and repression of histone synthesis during the cell cycle are regulated, in part, by modulating histone mRNA stability. When DNA synthesis stops, histone mRNA seems to be destabilized, perhaps via an autoregulatory circuit triggered by cytoplasmic histones. We have used an in vitro mRNA decay system to determine whether differential histone mRNA turnover is linked to changes in the basal activity of cytoplasmic mRNA-degrading enzymes. The basal level of the polysome-associated exonuclease enzyme or enzymes that degrade histone mRNA was similar in untreated cells and in cells exposed to DNA or protein synthesis inhibitors. Histone mRNA decay was accelerated in reactions supplemented with histones and soluble cytoplasmic factor(s) (S130), but S130s from control and inhibitor-treated cells were indistinguishable in these assays. The data indicate that basal exonuclease activity is stable or constitutive. The putative factor(s) required for autoregulating histone mRNA decay also do not change appreciably when DNA or protein synthesis is inhibited. The implications of these results with regard to the autoregulation of histone mRNA turnover are discussed. PMID- 2559296 TI - Location and molecular cloning of the structural gene for the deoxyguanosine triphosphate triphosphohydrolase of Escherichia coli. AB - The structural gene for deoxyguanosine triphosphate triphosphohydrolase (dGTPase) (EC 3.1.5.1) and its regulator, optA, have been located on a lambda phage carrying a 17.5kb Escherichia coli DNA insert. The DNA fragment has been excised and ligated into pBR325 and also transferred to another lambda vector. From the results of transduction and transformation experiments, we find that the structural gene for dGTPase is very closely linked to optA and dapD, which locates it at approximately 3.6 minutes on the genetic map of E. coli K12. We propose the mnemonic dgt as the designation for the structural gene for this enzyme. PMID- 2559297 TI - Transcriptional induction of Streptomyces cacaoi beta-lactamase by a beta-lactam compound. AB - The soil bacterium Streptomyces cacaoi produces an extracellular beta-lactamase. The beta-lactamase expression could be induced by the beta-lactam compound 6 amino penicillinoic acid (6-APA). In liquid cultures, a 50-fold increase in beta lactamase expression was observed within the first three hours after addition of 6-APA. Using the cloned beta-lactamase gene as a probe, it was shown that this increase was mediated at the level of transcriptional initiation. The start point of the induced beta-lactamase transcript was determined, and the nucleotide sequence of the promoter region was analysed. No noticeable homology was found to control regions of inducible beta-lactamase genes of other bacteria. A striking feature was the presence of six direct repeats (ten base pairs each) upstream of the promoter region. Thus, an example of an inducible regulatory gene system in this Gram-positive microorganism is presented. Also, the primary structure of the beta-lactamase was deduced, showing a high degree of homology with class A beta lactamases. PMID- 2559298 TI - Structure and function of hot spots providing signals for site-directed specific recombination and gene expression in Tn21 transposons. AB - Tn21- and Tn3-related transposons are widespread and carry various resistance determinants. The insertion points of different resistance genes were precisely defined in Tn2424, Tn1696, Tn2410, Tn4000 and its derivatives and compared to the corresponding sites in Tn7, pSA, R388, R46, Tn2603, Tn1331 and in Tn3-related elements. Insertional 'hot spots' located at the 3' end of different genes comprised 55 nucleotides and yielded more than 90% homology to the corresponding consensus sequence, termed hs1. Elements of this class were found to direct recA independent generation of deletions. Flanking the 5' ends, hs2 (CTAAAACAAAGTTA) comprised the terminal nucleotides of hs1. Functional properties of hot spots as recognition sites for site-specific recombination and regulation of gene expression indicate that they might be involved in transfer, stable inheritance and expression of prokaryotic genes. PMID- 2559300 TI - Families of bacterial signal-transducing proteins. AB - Bacteria can respond to a variety of environmental stimuli by means of systems generally composed of two proteins. The first protein (sensor or transmitter) is usually a transmembrane protein with cytoplasmic and extracytoplasmic domains. The extracytoplasmic domain (sensor) senses the environment and transfers the signal through the transmembrane domain to the cytoplasmic domain (transmitter), which has kinase activity. The second protein is located in the cytoplasm and contains an amino-terminal domain (receiver), which can be phosphorylated by the transmitter, and a carboxy-terminal region (regulator), which regulates gene expression by binding to DNA. The transmitter and receiver modules (the kinase and its target) are conserved in all signal-transducing systems and are the 'core structure' of this two-component system. The sensors and the regulators vary according to the stimuli they respond to and the DNA structure they interact with. On the basis of their sequence homology, the proteins belonging to such two component systems can be classified into different families, which are summarized in this review. PMID- 2559299 TI - pH-regulated gene expression in Salmonella: genetic analysis of aniG and cloning of the earA regulator. AB - The recently described aniG locus exhibits a series of unique regulatory features. The gene is exogenously coinduced by acid and D-mannose, its expression is maximal under anaerobiosis, and the system is regulated in an unusual manner by cyclic AMP. The external acid regulatory locus, earA, is a repressor protein that mediates the acid and mannose control of aniG. The earA locus was cloned and found to code for a 33K protein associated with membrane- and soluble fractions. A second locus, earB, was located immediately upstream from earA. The earB locus or its product interferes with the repression of aniG by EarA. Mutations in crp and cya were found to prevent transcription of aniG but only in an earA+ background. Analysis of an earA-cat fusion established that crp does not affect earA expression. While the physiological role of aniG/earA is unclear, this system serves as a model for external pH-regulated gene expression. The present data indicate that it is used to sense the presence of mannose in an acidic extracellular environment. This is particularly intriguing in that the system is not involved in the utilization of mannose as a carbon source. PMID- 2559301 TI - Synthesis of long, capped transcripts in vitro by SP6 and T7 RNA polymerases. PMID- 2559302 TI - Productive ascites growth of heterohybridomas between Epstein-Barr virus transformed human B cells and murine P3X63Ag8.653 myeloma cells. AB - Heterohybridomas between Epstein-Barr virus-transformed human B cells and murine myeloma P3X63Ag8.653 cells were found to produce sizable quantities of human monoclonal antibodies in nude mouse ascites. Mice injected intraperitoneally with two of such heterohybridomas yielded several milliters of ascites fluid which contained nearly 100-fold higher anti-platelet activities than those in routine culture supernatants. PMID- 2559303 TI - Association of collagenase and tissue inhibitor of metalloproteinases (TIMP) with hypertrophic cell enlargement in the growth plate. AB - In the transition from proliferating to hypertrophic cell zones in the growth plate, there is an increased in chondrocyte cell volume and a corresponding decrease in collagen content to allow for cell enlargement. To substantiate our hypothesis that collagenase is responsible for these changes, growth plates from rats treated with bisphosphonate (HEBP) were compared histologically and biochemically with growth plates from normal and vitamin D and phosphate deficient (-VDP) rats. HEBP-treated rats developed an expanded hypertrophic cell zone (HCZ) characterized by the presence of two distinct populations of hypertrophic cells. The proximal hypertrophic cells were only 2-fold enlarged compared to the proliferating cells, whereas 1/6 of the distal hypertrophic cells were enlarged almost 5-fold and appeared morphologically identical with hypertrophic cells from normal and -VDP rats. The HEBP growth plates were divided into cross-sectional thirds and analyzed for active and latent collagenase. The juxta-metaphyseal (lower 1/3) cartilage contained 100% of the fully enlarged hypertrophic cells and appeared identical to those found in normal and -VDP growth plates, along with 81% of the active and 77% of the total collagenase. Collagenase and tissue inhibitor of metalloproteinases (TIMP) were measured in extracts of similarly divided tissues. The presence of true collagenas was confirmed by using [3H]-telopeptide-free collagen. TIMP levels were inversely related to the presence of active collagenase and cellular hypertrophy. Substantial levels of latent collagenase were found in the extracellular fluid at sites of active collagenolysis, but not in the fluid phase surrounding the 2-fold enlarged hypertrophic cells. It is postulated that increased amounts of active collagenase and insufficient levels of TIMP may account for the reduced collagen content seen in the lower HCZ of both -VDP and HEBP rickets. Unlike active collagenase, which remains localized by binding to collagen, latent enzyme is probably restricted in its mobility throughout the extracellular space by diffusion, itself, or the interstices of the extracellular matrix. PMID- 2559304 TI - Expression of chloramphenicol acetyltransferase (CAT) from rodent type I collagen promoters in transfected osteosarcoma cells in vivo. AB - We examined the osteoblastic phenotype of permanently transfected ROS 17/2.8 cells in culture and in vivo, in order to evaluate their relevance for studies of the regulation of gene expression and gene function in osteoblastic cells. Recent reports indicate that the progeny transfected cells may substantially vary and differ from the parental cell line in their phenotype, particularly in their tumorigenicity. ROS 17/2.8 cells were transfected with genetic constructs expressing the CAT gene from either the rat alpha 1 (I) or the mouse alpha 2 (I) collagen promoters. Forty-four clonal cell lines display a range of CAT expression from the transfected collagen promoters in culture. Four of these cell lines were further characterized. Alkaline phosphatase activity in these four cell lines is higher than in fibroblastic cells. These four cell lines are tumorigenic in immunocompatible ACI rats and form calcified tumors similar to those formed by ROS 17/2.8. CAT expression could be demonstrated in tumor extracts of two of the four cell lines, which also expressed higher CAT levels in culture. We conclude that permanently transfected ROS 17/2.8 derived cell lines maintain their tumorigenicity and their osteoblastic-like phenotype, and thus may provide a useful system for studies of gene function and regulation in osteoblast like cells and bone-like tissue in vivo. PMID- 2559305 TI - Interstitial collagenase (MMP-1), gelatinase (MMP-2) and stromelysin (MMP-3) released by human fibroblasts cultured on acellular sarcoid granulomas (sarcoid matrix complex, SMC). AB - We studied collagenase, gelatinase and stromelysin syntheses by human fibroblasts cultured on three models of tridimensional matrix: native collagen sponge, native collagen complexed with glycosaminoglycans sponge, and acellular sarcoid matrix complex prepared from human sarcoid granulomas. Collagenase and stromelysin biosyntheses were differently stimulated according to culture conditions. Fibroblasts secreted a same amount of collagenase or stromelysin when cultured on collagen and collagen-glycosaminoglycans sponges, while collagenase and stromelysin secretions were widely amplified when cultured on sarcoid matrix complex. In contrast, gelatinase production was equally induced by the three culture conditions. In the different culture conditions on tridimensional matrix, the three matrix metalloproteinases were synthesized in a latent form. Thus, the sarcoid matrix complex stimulated the release of collagenase and stromelysin by fibroblast, but did not stimulate the release of gelatinase. This suggests that collagenase and stromelysin syntheses are co-regulated while gelatinase production is controlled by a distinct mechanism. PMID- 2559306 TI - Detection of human papillomavirus deoxyribonucleic acid in the female genital tract. AB - A total of 336 biopsies, scrapes and exfoliated cells from the cervix and from the lower genital tract were screened for human papilloma (HP) viral sequences of types 6, 11, 16 and 18 by Southern blot, dot blot and filter in situ (FISH) hybridizations with cloned 32P-radiolabeled HPV DNA probes. The specimens included cervical intraepithelial neoplasias (CIN I-III), carcinoma in situ and invasive carcinoma of the cervix and vagina, adenocarcinomas, vulvar and vaginal condylomata acuminata and healthy epithelial samples. The oncogenic HPV 16 was found in 46% of the cervical carcinomas. Most of the type 16 occurrences (75%) represented the third stage of inoperable cases. Similarly, HPV 18 was also most frequently present in this stage as well as in carcinoma in situ and in CIN III (25%, 18%). At the same time, in condylomata acuminata, types 6 and 11 were detectable in 88.7% of cares. In all, 13.5% of the normal samples harboured HPV DNA. PMID- 2559307 TI - Human tumor necrosis factor increases the resistance against Listeria infection in mice. AB - The resistance in mice against Listeria infection was augmented by treatment with recombinant human tumor necrosis factor (TNF). To elucidate this phenomenon, we examined the effect of TNF on macrophage activation. TNF-treated macrophages had listericidal activity in vitro and superoxide anion production. In addition, macrophage migration was inhibited in the presence of TNF. Therefore, activation of macrophages by TNF was similar to activation by macrophage-activating factor or macrophage-migration-inhibitory factor. PMID- 2559308 TI - Presence of group A and non-A rotaviruses in neonatal piglets in Campinas, SP, Brazil. AB - Rotaviruses were detected by polyacrylamide gel electrophoresis (PAGE) in 11 (84.6%) of 13 faecal specimens from neonatal piglets with acute diarrhoea in a piggery near the city of Campinas, State of Sao Paulo, Brazil. An immunoenzimatic assay for group A rotavirus (IEA-A) was positive in ten of the samples, all of which showed a PAGE profile typical of that group. Another sample was showed a group B profile in PAGE. An immunoenzimatic assay specific for group B (IEA-B) for this faecal sample was positive, confirming the PAGE results. PMID- 2559309 TI - [Electroencephalographic study of the effect of a benzodiazepine antagonist in hepatic encephalopathy]. AB - Observing animal models of fulminant hepatic failure lead to the hypothesis of a GABAergic origin of the comatose state in hepatic encephalopathy (HE). The hypothesis of hyperstimulation of gamma-aminobutyric acid-benzodiazepine (GABA BZ) receptors in HE has been tested with a BZ antagonist (flumazenil, Anexate) on 7 patients suffering from severe HE. The standard EEG has been recorded 30 min before and after slow IV perfusion of 1 mg flumazenil. Although we did not observe a complete EEG normalization, a significant improvement of EEG was observed after only a few minutes in 6 out of the 7 cases studied. Modification of the reactivity parallels clinical improvement of encephalopathy. These effects persist mostly 4 h after perfusion. The results are consistent with the hypothesis of hyperstimulation of GABA-BZ receptors in HE. The positive effect of flumazenil on vigilance level should encourage its use in chronic hepatic encephalopathy. PMID- 2559310 TI - Overview of the National Infant Mortality Surveillance (NIMS) project. AB - A slowdown in the decline of infant mortality in the United States and a continuing high risk of death among black infants (twice that of white infants) prompted a consortium of Public Health Service agencies, in collaboration with all states, to develop a national data base of linked birth and infant death certificates for the 1980 birth cohort. This project, referred to as National Infant Mortality Surveillance (NIMS), provides neonatal, postneonatal, and infant mortality risks for blacks, whites, and all races in 12 categories of birthweights. Tabulations were requested for infants born in single and multiple deliveries. For single-delivery births, tabulations included birthweight, age at death, race of infant, and each of these characteristics: infant's live-birth order, sex, gestation, type of delivery, and cause of death; and mother's age, education, prenatal care history, and number of prior fetal losses at greater than or equal to 20 weeks' gestation. An estimated 95% of eligible infant deaths were included in the NIMS tabulations. Analyses have focused on various components of infant mortality, including birthweight distribution of live births, neonatal mortality, and postneonatal mortality. The most important predictor for infant survival is birthweight; survival increases exponentially as birthweight increases to its optimal level. The nearly twofold higher risk of infant mortality among blacks than among whites was related to a higher prevalence of low birthweights, to higher mortality risks in the neonatal period for infants with birthweights of greater than or equal to 3,000 g, and to higher mortality during the postneonatal period for all infants, regardless of birthweight. Moreover, the black-white gap persisted for infants with birthweight of greater than or equal to 2,500 g, regardless of other infant or maternal risk factors. PMID- 2559311 TI - Isolation and molecular analysis of the maize P locus. AB - The maize P locus is involved in the synthesis of a red flavonoid pigment in the pericarp, cob and other floral tissues. The tissue-specific pattern of expression of certain P alleles suggests that P may be a complex locus, with more than one functional unit. The P-VV allele, which specifies variegated pericarp and variegated cob, however, shows that insertion and excision of the transposable element Ac affects both pericarp and cob expression as though cob and pericarp pigmentation are controlled by a single gene. Using Ac as a transposon tag, we have isolated 34 kb of genomic DNA from the P-VV and P-RR allele. The cloned DNA contains two 5.8 kb cross-hybridizing regions, in direct orientation relative to each other, separated by 6.6 kb of intervening DNA. A sequence motif of 250 bp is repeated at three locations within the cloned region: once within each of the 5.8 kb repeats, and once outside the 5.8 kb repeats. DNA fragments flanking the Ac element detect five transcripts in RNA from wild type (P-RR) that are absent from mutant (P-VV) tissues. To localize the transcribed sequences, DNA probes spanning the 34 kb of cloned DNA were used in Northern analysis of RNA from mutant and wild-type kernels. The results suggest the presence of a single transcriptional unit located primarily within the DNA between the 5.8 kb repeats. The five RNAs transcribed from this region may be formed by alternative splicing. The size of the P gene derived from the length of the transcribed region seems much smaller than the gene size estimated from Ac-induced P-VV mutations. PMID- 2559313 TI - Mobile elements and transposition events in the cut locus of Drosophila melanogaster. AB - We have cloned from the Oregon R strain of Drosophila melanogaster a 240 kb segment of DNA that contains the cut (ct) locus, and characterized the region for the presence of repetitive elements. Within this region at least five copies of the suffix element were detected, as well as several putatively novel mobile elements. A number of mutations obtained from the unstable ctMR2 strain and its derivatives were mapped within the cut locus. Comparison between parental and daughter strains indicates that frequently two or more independent transposition events involving the cut locus occur simultaneously within a single germ cell, thus providing a molecular basis for the transposition explosion phenomenon. PMID- 2559312 TI - Identification of a regulatory nifA type gene and physical mapping of cloned new nif regions of Azospirillum brasilense. AB - Three new Tn5-mutagenized nif genes of Azospirillum brasilense were characterized. The sizes of the restriction fragments and the restriction maps of the cloned nif DNA regions showed that these nif genes are distinct from those reported earlier, e.g. nifHDK, nifE, nifUS, fixABC. The Nif27 mutant was identified as a nifA type regulatory gene of A. brasilense (a) by genetic complementation with nifA of Klebsiella pneumoniae, (b) by the absence of nitrogenase iron protein in western protein blots and (c) by its inability to activate expression of a nifH-lacZ fusion. The growth characteristics of the three mutants showed that none of them is defective in general nitrogen regulatory (ntr) genes. Also, no homology was detected between the three nif DNA regions of the mutants, cloned in pMS188, pMS189 and pMS197, and the K. pneumoniae nif, glnA or ntr genes. In addition, the fixABC genes of Bradyrhizobium japonicum did not show any hybridization with the cloned Azospirillum genes. Unlike the situation in enteric bacteria, the nif genes in A. brasilense are scattered and span a region of about 65 kb. PMID- 2559314 TI - FemA, a host-mediated factor essential for methicillin resistance in Staphylococcus aureus: molecular cloning and characterization. AB - The methicillin resistance determinant (mec) in Staphylococcus aureus resides on additional DNA not present in isogenic sensitive cells. However, besides mec, other chromosomally determined factors are essential for expression of methicillin resistance. We cloned and characterized a chromosomally determined gene which encodes a factor essential for the expression of methicillin resistance (femA) in S. aureus. femA mapped in chromosomal segment number 18, genetically very distant from the methicillin resistance determinant (mec). The product of femA was a protein of an apparent size of 48 kDa. FemA restored methicillin resistance in S. aureus that had become sensitive to methicillin by insertion of omega 2003 (femA::Tn551). Although FemA was needed for cell growth in the presence of beta-lactam antibiotics, it had no influence on the synthesis of the low affinity, additional penicillin-binding protein (PBP2') encoded by mec and known to be essential for cell wall synthesis in the presence of inhibitory concentrations of methicillin. Nucleotide sequence analysis, Northern RNA blotting and S1 nuclease RNA mapping suggested that femA was transcribed on a polycistronic mRNA. This mRNA contained the coding region (ORF419) producing a protein of 47 kDa. The nucleotide and amino acid sequence of FemA showed homologies with ORF419, suggesting that these genes arose by gene duplication. In addition we present evidence for a second chromosomal factor, femB, involved in expression of methicillin resistance which maps close to femA. PMID- 2559315 TI - Double Ds elements are involved in specific chromosome breakage. AB - The structure of the unstable Ds-induced sh-m5933 allele of the maize sucrose synthase gene was analysed and a double Ds structure found in opposite orientation on both sides of a 30 kb insert interrupting the sucrose synthase gene. The double Ds structures bordering the insert are identical over a distance of approximately 3 kb. These double Ds structures and the DNA segments beyond them are in opposite orientation and identical over a distance of approx. 5.3 kb. A hypothesis for how such a symmetrical structure could be formed is proposed. When one complete Ds element was excised from one of the double Ds structures a half Ds element was left behind. This half Ds element was found in one revertant strain which displayed an altered pattern of chromosome breakage compared to revertant strains which had not undergone Ds excision. Nine new maize strains which showed a similarly altered chromosome breakage pattern were isolated. In all nine cases we observed an indistinguishable deletion in the genomic DNA. These excisions are likely to be the result of similar excision events to that described above. We conclude that double Ds structures are responsible for Ds induced chromosome breakage. PMID- 2559316 TI - New Escherichia coli gyrA and gyrB mutations which have a graded effect on DNA supercoiling. AB - We isolated new gyrA and gyrB mutations in Escherichia coli which have a graded effect on DNA supercoiling. The mutants, selected respectively for resistance to nalidixic acid and coumermycin, were sorted by means of a rapid in vivo assay of DNA gyrase activity (Aleixandre and Blanco 1987). Cells carrying a gyrB (Cour) mutation usually showed a decrease in DNA supercoiling, which would indicate a reduction in gyrase activity. In contrast, most of the gyrA (Nalr) mutations had no significant effect on DNA supercoiling. Moreover, they conferred a high level of resistance to nalidixic acid and other quinolones, thus being similar to the gyrA (Nalr) mutants currently used. We also detected rare gyrA mutants showing a reduction in DNA gyrase activity. These mutants were, in addition, resistant to only low concentrations of quinolones, which allowed us to use the phenotype of partial quinolone resistance as an indicator to score gyrA mutations affecting DNA supercoiling. When gyrB mutations were introduced into the gyrA mutants, these became more sensitive to quinolones and a decrease in supercoiling was observed. Moreover, the topA10 mutation sensitized gyrA (Nalr) cells to quinolones. We conclude therefore that the GyrA-dependent quinolone resistance is diminished as a consequence of the reduction either in topoisomerase I or gyrase activities. PMID- 2559317 TI - Tagging of a maize gene involved in kernel development by an activated Uq transposable element. AB - A quiescent Uq transposable element has been activated in a maize plant treated with 5-aza-2'-deoxycytidine. This activated Uq cosegregates with a heritable dominant miniature (Mn) kernel phenotype, indicating its physical association with a maize miniature locus (Mn::Uq). The Mn::Uq mutant is dominant in producing a miniature seed phenotype of variable size and in reducing seedling vigor in the early growth stage. Genetic experiments indicate that the Mn::Uq mutant also affects the activity of the male gametophyte, whereby pollen germination is inhibited, thus lacking pollen tube growth resulting in the male nontransmissibility of this mutant. Proof for the Uq element in this mutant is derived by its ability to transactivate the standard a-ruq reporter allele to yield spotted aleurone tissue. However, the Mn::Uq mutant does not transactivate a normally Uq-responsive c-ruq allele, suggesting a structural difference between the two ruq receptors at the A1 and C1 loci. It is anticipated that cloning of the Uq transposable element would facilitate the molecular cloning and characterization of the maize miniature gene. PMID- 2559319 TI - Identification of human picornaviruses by nucleic acid probes. AB - Human picornaviruses include rhinoviruses and enteroviruses which are responsible for both common and severe clinical diseases. Rhinoviruses are a frequent cause of respiratory infections while members of enterovirus subgroups, polio, coxsackie and ECHO viruses are often responsible for infections of the central nervous system, myocarditis, myositis etc. Human picornaviruses consist of nearly two hundred serotypes and therefore their specific identification after virus isolation, or the diagnosis based on the detection of immune response in patients, is problematic and does not usually provide virological diagnosis at the acute phase of illness. New methods for detection of picornavirus genomic RNA together with increasing knowledge of the nucleotide sequences of this virus group offer interesting possibilities for diagnostic procedures. Spot hybridization, in situ hybridization and enzymatic amplification of specific sequences have successfully been used for this purpose. Probes covering the 5' non-coding part of the genome, and also sequences derived from the region coding for non-structural proteins, can be used as broadly reacting reagents in picornavirus detection. Specific sequences are mainly found in the capsid protein region of the genome. cDNA probes and synthetic oligonucleotides are useful in rapid identification of picornaviruses after amplification in cell cultures and in epidemiological analysis. The biochemical amplification methods may enable recognition of picornaviruses directly in clinical samples in the near future. In situ hybridization methods have been of special interest because they can be used to reveal the presence of enterovirus genomes in biopsy specimens from e.g. affected heart muscle in patients with myocarditis and cardiomyopathy. PMID- 2559318 TI - Conserved function in Nicotiana tabacum of a single Drosophila hsp70 promoter heat shock element when fused to a minimal T-DNA promoter. AB - To demonstrate the extent of evolutionary conservation in the mechanism of induction of heat shock genes between plants and animals, the minimal sequence from the Drosophila hsp70 promoter sufficient to confer heat shock inducible transcription in tobacco was determined. Segments of the hsp70 promoter were fused to a minimal promoter of the T-DNA indole-3-acetamide hydrolase (iaaH) gene, in a chimaeric gene fusion to a neomycin phosphotransferase (NPT II) reporter gene. Sequences bearing one or more heat shock elements (HSEs) rendered the minimal promoter heat shock inducible, with a 37 bp fragment containing a single complete HSE sufficing. The induced NPT II mRNA peaked during the heat shock period, but the maximal level of NPT II activity was not observed until 4 h later in the recovery phase, showing that the translation of the NPT II mRNA was shifted from the heat shock period of the recovery phase. That similar sequences containing a single HSE of the Drosophila hsp70 promoter could function in both flies and tobacco indicates the high degree of homology between the two heat shock gene induction systems. PMID- 2559321 TI - Solution hybridization and enzyme immunoassay for biotinylated DNA-RNA hybrids to detect enteroviral RNA in cell culture. AB - A non-isotopic hybridization assay is described for detection of enteroviral RNA in cell culture. Two biotin-labelled cDNA probes, corresponding to 1 kb from the 5' end and 3.5 kb from the 3' end of the coxsackievirus B3 genome, were hybridized in solution with protease and detergent-treated cell culture suspensions. Labelled DNA-RNA hybrids were captured on microtiter plates coated with anti-biotin antibody and bound hybrids were measured with a beta galactosidase-labelled monoclonal antibody specific for DNA-RNA hybrids. Coxsackie B3 was detected at a concentration of 500 pfu ml-1. The limit of detection for other enteroviruses ranged from 10(3.3) to 10(5.8) pfu ml-1. The enteroviruses that could be detected included coxsackie B1 and 3, coxsackie A1-6 and 15, poliovirus types 1-3, and enteroviruses 7, 11, and 71. ECHO 22 was the only enterovirus, of those that were tested, that could not be detected. The solution hybridization reaction and enzyme immunoassay for DNA-RNA hybrids does not require the use of radiolabelled probes or extraction of RNA with phenol. The assay yields a quantitative endpoint, which avoids the subjectivity inherent in membrane-based methods. These features would make the assay more adaptable to clinical laboratories than other formats which have been devised for measurement of viral RNA. PMID- 2559320 TI - Rapid detection of herpes simplex virus DNA by in situ hybridization with photobiotin-labelled double-stranded DNA probes. AB - An assay for rapid detection of herpes simplex virus in infected cells is described. The assay utilizes in situ hybridization with photobiotin-labelled double-stranded DNA probes prepared from HSV-1 DNA cloned in plasmid vectors. The assay provided an alternative method for earlier detection of virus in cell cultures with the ease of preparation of photobiotin-labelled double-stranded DNA. PMID- 2559322 TI - Identification of rotaviruses by dot-blot hybridization using an alkaline phosphatase-conjugated synthetic oligonucleotide probe. AB - We have evaluated a recently-developed dot-blot hybridization assay for the detection of human rotaviruses using an alkaline-phosphatase conjugated oligonucleotide probe. The lower detection limit of this assay was 1 ng (approximately 5 x 10(7) copies) of the double-stranded (ds) RNA, when a purified preparation from serotype 1 human rotavirus was used but appeared to be much higher when applied on clinical specimens. This assay could detect dsRNA from rotavirus strains belonging to serotypes 1 through 6, 8 and 9. A total of 235 stool specimens were used to evaluate the oligonucleotide probe assay in comparison with polyacrylamide gel electrophoresis and latex agglutination assay (Rotalex). When polyacrylamide gel electrophoresis was used as a gold standard, sensitivity, specificity and positive and negative predictive values of the probe assay were 84, 100, 100 and 89%, respectively. These values were slightly better than those of Rotalex assay which is commonly used in clinical laboratories in Japan. Although the probe assay requires more hands-on time than the immunoassays, the high specificity of this probe assay recommends it as a confirmatory test in the clinical laboratory setting. PMID- 2559323 TI - Polybrominated naphthalene and diiodobenzene interactions with specific binding sites for 2,3,7,8-tetrachlorodibenzo-p-dioxin in rat liver cytosol. AB - We provide evidence for two new classes of halogenated aromatic hydrocarbon ligands for the 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD or Ah) receptor: brominated naphthalenes and iodobenzenes. Polybrominated naphthalenes with four or more bromine atoms concentrated in lateral positions were shown to bind specifically and with high affinity (Kd approximately 10(-8) M) to the Ah receptor in rat liver cytosol preparations. The hexabrominated naphthalene isomers bind with high and nearly equal affinities but have been previously shown to have different toxicological properties. Possible explanations for these differences include differences in metabolism, antagonist versus agonist Ah receptor binding of some isomers, and the involvement of other binding sites in vivo that require different structural requirements. The moderate binding activity of the diiodobenzenes suggests that thyroid hormones should receive further study as possible endogenous ligands for the Ah receptor. It is difficult to explain the binding results with these two classes of compounds using previously developed molecular concepts for Ah receptor interactions based primarily on molecular size considerations. PMID- 2559324 TI - [Cloning and analysis of the primary structure of an element, related to the murine mammary cancer virus, from the genome of the Djungarian hamster]. AB - 11 recombinant bacteriophages from the genomic library of Djungarian hamster genome, that carry MMTV-related sequences (MRS-Ps), have been cloned with the murine mammary cancer virus (MMTV) as a hybridization probe. The sequences are repeated 50 times in the genome. MRS-Ps contain the tracts of homology with the long end repeat MMTV, the genes pol and, possibly, env, but not with the gag gene. Sequencing of the 0,87 kb restrict, common to all EcoRI-BamHI clones, and the analysis of the sequence have demonstrated the high level of homology with the pol gene of MMTV and its origination from the somewhat functioning gene. PMID- 2559325 TI - [The effect of various rec-mutations on the frequency of the Tn917 transposition in Bacillus subtilis cells]. AB - The effect of 11 rec-genes on the transposition frequency of Tn917 has been studied. Transposition frequencies in RecP, RecF15, RecB3 mutants differed from the ones in the control strains. The collection of mitomycin-sensitive mutants has been tested for transposition proficiency. The mms315 mutation decreasing the transposition frequency possess the properties of the rec mutation too. PMID- 2559327 TI - Enhancement of 5'-nucleotidase activity of human leukemic cells after fractionation: implications for cancer and aging. AB - Previous studies reported that 5'-nucleotidase activity was undetectable or at much lower levels in the homogenate of human chronic lymphocytic leukemic (CCL) cells than in normal lymphocytes. In the present study, 5'-nucleotidase specific activity in acute myelocytic leukemia (AML), which varied in a range from undetectable to 1.4 (nmoles/min.mg protein), was enhanced by cell fractionation, from undetectable in the homogenate, up to 18.8 +/- 1.2, 6.4 +/- 0.7 and 0.68 +/- 0.12 in plasma membranes, microsomes, and cytosol fraction, respectively. In a further fractionation of the cytosol of various leukemic cells with ammonium sulfate, 5'-nucleotidase specific activity increased up to 14-fold in the 60% (NH4)2SO4 fraction, with a recovery of 1266 +/- 115%. These data suggest that 5' nucleotidase activity in fractionated leukemic cells is higher than reported previously and that the sum of 5'-nucleotidase activity in subcellular compartments is higher than that detected in the homogenate. Furthermore, even when 5'-nucleotidase was undetectable in a homogenate, it became detectable in the plasma membranes, suggesting that its ecto-enzyme function is still active in leukemic cells. The undetectable or low 5'-nucleotidase in the homogenate is indicative of (1) the enzyme itself being in an inactive form but becoming active after the fractionations, or (2) the presence of a factor(s) that prevents the enzyme from being detected but that is separated from the enzyme by the fractionations. In both cases, the rate of nucleotide catabolism by inactive 5' nucleotidase in rapidly proliferating leukemic cells should be slower than when the enzyme is active. The present finding is consistent with our previous findings that during normal cell aging the high 5'-nucleotidase activity is associated with senescent non-proliferating cells but low or undetectable activity with rapidly proliferating immortal cells. The implications of 5' nucleotidase for DNA synthesis in aging and cancer are discussed. PMID- 2559326 TI - Age-dependent increase of DNA topoisomerase II activity in quail oviduct; modulation of the nuclear matrix-associated enzyme activity by protein phosphorylation and poly(ADP-ribosyl)ation. AB - Nuclear DNA topoisomerase II activity in quail oviduct tissue was found to increase by about 70% with age. This age-dependent increase was observed with both the enzyme in whole nuclear extract and nuclear matrix-associated topoisomerase II. Both purified topoisomerase II and the nuclear matrix-bound enzyme were found to be modifiable by phosphorylation and poly(ADP-ribosyl)ation. Phosphorylation of the purified enzyme by isolated nuclear protein kinase NII or protein kinase C resulted in a 2- to 3-fold increase in specific activity, while poly(ADP-ribosyl)ation by soluble poly(ADP-ribose) synthetase caused a 50% inhibition of the enzyme. Using immunoprecipitation and immunoblotting procedures, phosphorylation and poly(ADP-ribosyl)ation could also be demonstrated to occur with the nuclear matrix-associated enzyme. The nuclear matrix-associated NII-like protein kinase activity, assumed to be involved in post-translational modification of topoisomerase II, displayed a 1.4- to 1.6-fold increase in old animals compared to mature ones, while the matrix-bound poly(ADP-ribose) synthetase activity decreased by about 50%. It is suggested that age-correlated enhancement of DNA topoisomerase II activity, possibly due to age-dependent changes in activities of nuclear protein kinases and poly(ADP-ribose) synthetase, may result in alterations in the topological state of DNA, possibly affecting DNA replication, transcription and repair with age. PMID- 2559328 TI - A polyubiquitin gene from Trypanosoma brucei. PMID- 2559329 TI - Entamoeba histolytica: molecules involved in the target cell-parasite relationship. AB - To detect molecules of Entamoeba histolytica involved in the trophozoite-target cell interaction, three different antisera were generated: (a) two rabbit antisera, one against total amebic proteins and another directed specifically to the 112-kDa adhesin; and (b) a mouse antiserum against amebic molecules adhering to the red blood cell (RBC) surface after incubation of RBCs with total soluble protein from trophozoites (anti-adhesion serum). All three antisera recognized the 112-kDa adhesion. Adhesion of this molecule to the RBC surface was temperature-dependent. More of the 112-kDa adhesion was found on the surface of RBCs incubated with trophozoites at 37 degrees C than on RBCs incubated at room temperature or at 0 degree C. Experiments using both anti-adhesin and anti-total ambebic protein sera revealed the presence of 210, 160, 112, 90, 70, 50, and 24 kDa proteins on RBC incubated with trophozoites. Surface proteins obtained from iodinated MDCK cells recognized amebic proteins of 112, 90, and 48-50 kDa. Virulence-deficient mutants presented a similar amount of the 112-kDa adhesin to the wild-type strain. However, in mutants, the adhesion was not functional, since they did not adhere to RBCs. 90- and 24-kDa proteins were also found to be altered in mutants. PMID- 2559330 TI - Fungispecificity of fluconazole against Candida albicans. AB - Candida albicans is an opportunistic pathogen of human mucosal surfaces. Colonization of oral and vaginal mucosa by this yeast is antagonized by the resident normal bacterial population. However, antibacterial therapy can alter the normal flora to allow fungal cells to attach, grow and invade host tissues. We studied the antimicrobic activity of fluconazole against clinical isolates of oral and vaginal bacteria and Candida albicans in vitro and in vivo by scanning and transmission electron microscopy; we also compared the bactericidal activity of fluconazole with clotrimazole in vitro by microbiologic assay. Fluconazole lysed fungi but did not change the ultrastructure of bacteria. Clotrimazole, but not fluconazole, was bactericidal against lactobacillus and streptococcus, the principal species of the oral and vaginal cavities. We conclude that Candida albicans, but not oral and vaginal bacteria, is susceptible to fluconazole. These observations help explain the antimycotic specificity of fluconazole and its efficacy against candidiasis in humans. PMID- 2559331 TI - Inhibition of neuronal noradrenaline transport (uptake1) and desipramine binding by amiloride and ethylisopropylamiloride. AB - The diuretic amiloride and its N-5 substituted analogue ethylisopropylamiloride (EIPA) inhibit both the specific high affinity desipramine binding to isolated plasma membranes of PC12 rat phaeochromocytoma cells and the carrier-mediated neuronal uptake of noradrenaline into PC12 cells. The inhibition by EIPA of both desipramine binding (Ki = 5.6 mumol/l) and noradrenaline uptake (Ki = 24 mumol/l) inversely depend on the extracellular sodium concentration. The degree of inhibition increased with decreasing sodium concentration. A more detailed analysis of the mode of interaction revealed a competitive interaction between EIPA and desipramine binding but an "uncompetitive" interaction between EIPA and noradrenaline uptake. EIPA is the first inhibitor of uptake1 known so far, which reduces both Km and Vmax of neuronal noradrenaline transport. Extracellular alkalinization from pH 7.4 to 7.9 during incubation with EIPA markedly increased the effects on the kinetics of noradrenaline transport. A model has been proposed to explain the kinetic phenomena. It is based on the hypothesis that EIPA diffuses through the plasma membrane and binds to the inward facing sodium binding site of the neuronal noradrenaline carrier. PMID- 2559332 TI - The extent of neuronal re-uptake of 3H-noradrenaline in isolated vasa deferentia and atria of the rat. AB - After pretreatment of rats with reserpine and pargyline (to inhibit vesicular uptake and monoamine oxidase, respectively) and after inhibition of catechol-O methyl transferase (by U-0521) and in calcium-free solution, the adrenergic neurones of isolated vasa deferentia and atria were loaded with 3H-noradrenaline. The spontaneous efflux of 3H-noradrenaline and 3H-dihydroxyphenylglycol was determined, as well as the steady-state effect of two concentrations of desipramine. On the basis of a mathematical model of the adrenergic nerve ending, fractional rates (FR = rate of flux divided by tissue tritium content) were calculated for unidirectional outward diffusion, for outward transport and for neuronal re-uptake (all for 3H-noradrenaline). Although the density of adrenergic innervation is lower in atria than in vasa deferentia, neuronal re-uptake amounted to about 90% of the spontaneous efflux of 3H-noradrenaline in both tissues. While the FR for unidirectional outward diffusion was virtually the same in both tissues, the FR for outward transport of 3H-noradrenaline was more than three times higher in atria than in vasa deferentia. There is, as yet, no explanation for this pronounced difference. PMID- 2559333 TI - Inhibition by nucleotides acting at presynaptic P2-receptors of sympathetic neuro effector transmission in the mouse isolated vas deferens. AB - Effects of nucleotides and nucleosides on smooth muscle tension and the release of previously stored [3H]-noradrenaline were studied in the mouse isolated vas deferens. The tissue was stimulated twice by 20 electrical field pulses delivered at 2 Hz (S1, S2). alpha,beta-Methylene-ATP, ATP gamma S, ATP and UTP elicited contraction, with potency decreasing in that order; there was no contractile response to adenosine (up to 100 mumol/l) and uridine (up to 1 mmol/l). The electrically evoked overflow of tritium was reduced by the drugs in the following order of potency: ATP gamma S greater than ATP = adenosine greater than UTP; alpha,beta-methylene-ATP (up to 10 mumol/l) and uridine (up to 1 mmol/l) did not significantly change the evoked overflow. 8-(p-Sulphophenyl)theophylline did not alter the contractile responses to the nucleotides; it prevented the overflow inhibiting effect of adenosine and reduced that of UTP; the overflow-inhibiting effects of ATP and ATP gamma S were not significantly attenuated. After prolonged exposure to alpha,beta-methylene-ATP, all contractile nucleotide effects were abolished; in contrast, the depression by adenosine and the nucleotides of the evoked overflow of tritium persisted. None of the effects was changed by indometacin, yohimbine or reactive blue 2. It is concluded that ATP, ATP gamma S, alpha,beta-methylene-ATP and UTP produce contraction of the vas deferens by activation of P2x-receptors. Moreover, the nucleotides inhibit per se the release of [3H]-noradrenaline (and presumably the co-transmitter mixture of noradrenaline and ATP); the effect of ATP is not, or only to a small extent, due to breakdown to adenosine. The presynaptic site of action of the purine nucleotides is a P2 receptor which differs from the P2x-receptor and may be a reactive blue 2 resistant "P2y-like" receptor. PMID- 2559335 TI - A novel type of GABA receptor in rat spinal cord? AB - The depolarization-evoked release of gamma-aminobutyric acid (GABA) and its possible modulation mediated by autoreceptors were studied in nerve endings isolated from rat spinal cord and prelabeled with the radioactive aminoacid. In the presence of the GABA uptake inhibitor SK&F 89976A [N-(4,4-diphenyl-3-butenyl) nipecotic acid], used to minimize carrier-mediated homoexchange, exogenous GABA (1-10 mumol/l) decreased in a concentration-dependent way the release of 3H-GABA evoked by 15 mmol/l KCl. The GABAA receptor agonist muscimol (10-100 mumol/l) did not affect the K+ (15 mmol/l)-evoked 3H-GABA release. Similarly ineffective was the GABAB receptor agonist (-)-baclofen (3-100 mumol/l). The effect of GABA was not counteracted by the GABAA receptor antagonists bicuculline,picrotoxin or SR95531 [2-(3'-carbethoxy-2'-propenyl)-3-amino-6-paramethoxy-phenyl-pyr idazinium bromide].(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559334 TI - Estimation of pA2 values at presynaptic alpha 2-autoreceptors in rabbit and rat brain cortex in the absence of autoinhibition. AB - An attempt was made to determine pA2 values of antagonists at the presynaptic, release-inhibiting alpha 2-autoreceptors of rabbit and rat brain cortex under conditions when there was very little released noradrenaline in the autoreceptor biophase and, hence, pA2 values were not distorted by endogenous autoinhibition. Cortex slices were preincubated with 3H-noradrenaline and then superfused and stimulated by trains of 4 pulses delivered at 100 Hz or, in a few cases, by trains of 36 pulses at 3 Hz. The alpha-adrenoceptor agonists clonidine, noradrenaline, and alpha-methylnoradrenaline concentration-dependently decreased the stimulation-evoked overflow of tritium. The alpha-adrenoceptor antagonists yohimbine, rauwolscine and idazoxan did not increase the overflow of tritium elicited by 4 pulses/100 Hz in rabbit brain slices and increased it only slightly in rat brain slices. In contrast, the antagonists increased markedly the overflow at 36 pulses/3 Hz. All antagonists caused parallel shifts to the right of the concentration-response curves of clonidine, noradrenaline, and alpha methylnoradrenaline. pA2 values were calculated either from linear regression of log [agonist concentration ratio - 1] on log [antagonist concentration] or from sigmoid curve fitting. The slopes of the linear regression lines were close to unity, and the pA2 values calculated by the two methods agreed well. There was no consistent preferential antagonism of any antagonist to any agonist. pA2 values determined with stimulation by 4 pulses/100 Hz were by 0.53-0.80 log units higher than those determined with stimulation by 36 pulses/3 Hz.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559336 TI - Histamine inhibits activation of human neutrophils and HL-60 leukemic cells via H2-receptors. AB - The effects of prostaglandin E1 (PGE1) and histamine on activation of superoxide (O2-) formation, exocytosis of beta-glucuronidase and aggregation in human neutrophils and HL-60 leukemic cells were studied. PGE1, histamine and impromidine, a potent H2-agonist, inhibited O2- formation in neutrophils induced by the chemotactic peptide, N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMet Leu-Phe) with IC50 values of 0.5 microM, 8 microM and 2 microM, respectively. The full H1-agonist and weak partial H2-agonist, betahistine, was much less potent and effective than histamine. Dibutyryl cyclic AMP and forskolin mimicked the effects of histamine and PGE1 on O2- formation. The H2-antagonist, famotidine, competitively reversed histamine-induced inhibition of O2- formation with a pA2 value of 7.5. Histamine inhibited O2- formation when added prior to or after fMet Leu-Phe. fMet-Leu-Phe-induced aggregation and release of beta-glucuronidase in neutrophils were less sensitive to inhibition by PGE1, histamine, dibutyryl cyclic AMP and forskolin than O2- formation. The inhibitor of cyclic AMP-specific phosphodiesterase, rac-4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone (Ro 20 1724), additively enhanced the inhibitory effects of histamine and PGE1 on the above cell functions. In HL-60 cells differentiated by dimethyl sulfoxide or dibutyryl cyclic AMP, histamine, impromidine and PGE1 but not betahistine inhibited fMet-Leu-Phe-induced O2- formation as well. Our data suggest that histamine inhibits activation of neutrophils and HL-60 cells via H2-receptors through activation of adenylyl cyclase and increased formation of cyclic AMP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559337 TI - Effects of adenosine analogues on contractile response and cAMP content in guinea pig isolated ventricular myocytes. AB - In the present study the effects of adenosine analogues were investigated on cAMP content and contractile response in guinea-pig ventricular myocytes. The adenosine analogues (-)-N6-phenylisopropyladenosine (R-PIA), 5'-N ethylcarboxamideadenosine (NECA) and (+)-N6-phenylisopropyladenosine (S-PIA) in the presence of 0.01 mumol/l isoprenaline reduced contractile response concentration-dependently. R-PIA and NECA were about equipotent (IC25: 0.01 mumol/l and 0.039 mumol/l respectively), while S-PIA was less potent (IC25: 0.6 mumol/l). Isoprenaline stimulated cAMP content was reduced by R-PIA (IC25: 0.004 mumol/l) and with lower potency by S-PIA (IC25: 0.15 mumol/l), but the extent of reduction of cAMP by R-PIA and S-PIA (to 55% and 64% respectively) was less than the reduction of contractile response (to 26% and 55% respectively). This suggests that the effects of R- and S-PIA on contractile response are only in part due to a reduction in cAMP content. In addition, NECA did not decrease cAMP content but decreased contractile response to the same extent as R-PIA. Similar results were obtained in the presence of the phosphodiesterase inhibitor Ro 20 1724. Time course studies revealed that the effects of R-PIA (1 mumol/l) on cAMP content and contractile response coincided reaching steady state after 5 min and remained stable thereafter. The effects of NECA (1 mumol/l) on contractility also reached steady state within 5 min, whereas it did not change cAMP content. It is concluded that the reduction of contractility by adenosine analogues in the presence of isoprenaline can only in part be explained by a reduction of cAMP content.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559338 TI - Human cells of neural origin are permissive for bovine leukemia virus. AB - Bovine leukemia virus (BLV) propagated in a cell clone of fetal lamb kidney origin was transmitted by cell contact to different mammalian cells including human cells. The transmission of the BLV genome was effectively achieved by cocultivation of mitomycin-C-killed, virus-producing cells of the cell clone with recipient cells. In particular, human cells of neural origin were highly susceptible to BLV infection, while some other cells were resistant. The transmission of the BVL genome from virus-nonproducing cells failed which suggests the existence of virus specific receptors on the cells. The donor cells contained three integrated BLV proviruses. In recipient cells only one provirus was found. The majority of cells contained both unintegrated and integrated BLV provirus. In the cells containing the transmitted BLV, the viral genome was expressed to its protein products. The results indirectly suggest that retroviruses with similar properties could cause various neural diseases in man. PMID- 2559339 TI - Early senile plaques in Down's syndrome brains show a close relationship with cell bodies of neurons. AB - A sensitive methenamine silver/Nissl stain was used to study the morphology and relationship of pre-plaques (presumed early senile plaques) in Down's syndrome brains to glial nuclei, capillaries and neuronal perikarya. The larger pre plaques (greater than 50 microns) usually encompassed all of these tissue elements. However, the smaller pre-plaques (less than or equal to 50 microns) were almost always found immediately adjacent to, or around the cell bodies of neurons (often with associated satellite cells), and they failed to show any consistent, close spatial relationship to the other tissue components. Thus we consider an early stage of pre-plaque formation to be the deposition of amyloid adjacent to the cell body of a morphologically normal neuron. Based on the study of transitional forms, we suggest that the amyloid progressively accumulates around the cell body until the enclosed neuron degenerates. How these pre-plaque lesions might eventually develop into the typical plaque structure is uncertain. Our observations support the theory of a neuronal origin for plaque amyloid. PMID- 2559340 TI - Effect of interleukin-1 on ACTH and corticosterone secretion in dexamethasone and adrenalectomized pretreated male rats. AB - The present study was designed to investigate the role of glucocorticoids in mediating the stimulatory effect of interleukin-1 (IL-1) upon the adrenocortical (AC) axis. Intact male rats were injected with either vehicle or dexamethasone (DEX) (5-50 micrograms/100 g b.w.) and were subsequently exposed to ether stress or to an intracerebroventricular (i.c.v.) injection of 2 U recombinant human IL-1 beta. In addition, adrenalectomized (ADX) rats were injected i.c.v. with 2U IL-1; for comparison ADX rats were also exposed to ether stress and insulin-induced hypoglycemia. In intact rats, ether-stress-induced AC activation was much more sensitive to the DEX inhibitory effect than the IL-1-induced AC activation. In ADX animals, insulin-induced hypoglycemia and ether stress produced a significant increase in ACTH serum values whereas IL-1 administration failed to activate ACTH secretion. Glucocorticoid binding studies showed that an i.c.v. injection of 2 U IL-1, 3 h before sacrifice, markedly reduced the specific in vitro binding of 3H corticosterone by the nuclear fraction of dorsal hippocampal tissue slices. These results suggest that IL-1 may activate the AC axis, at least in part, by interfering with the negative feedback effect of circulating glucocorticoids. PMID- 2559341 TI - Diurnal rhythmicity of beta-1- and beta-2-adrenergic receptors in ovariectomized, ovariectomized estradiol-treated and proestrous rats. AB - The effect of norepinephrine on LH is complex: the ovarian steroidal milieu appears to determine whether norepinephrine stimulates or inhibits LH secretion. It has been proposed that steroids allow norepinephrine to have opposite effects on LH by altering the relative concentrations of alpha 1-(stimulatory) and beta (inhibitory) adrenergic receptors in the hypothalamus. Thus, many investigators have argued that estradiol may permit norepinephrine to stimulate LH release by increasing the density of alpha 1- and decreasing the density of beta-adrenergic receptors in one or more key hypothalamic regions. To test this hypothesis specifically, we measured the density of beta 1- and beta 2-adrenergic receptor densities in proestrous, ovariectomized, and ovariectomized estradiol-treated rats at various times of day to determine (1) whether the densities of beta receptors exhibit diurnal rhythmicity, (2) whether beta-receptors decrease during the time of increased LH secretion and/or (3) how steroidal milieu influences the density and/or the rhythm of receptor densities. The densities of beta 1- and beta 2-receptors exhibit a diurnal rhythm in some brain areas. These rhythms are detectable only in proestrous and ovariectomized rats and only in selected brain regions. Estrogen treatment has opposing effects in different brain regions. It suppresses the rhythm of beta 1-receptor concentrations in ovariectomized rats and also suppresses the average density of receptors in the suprachiasmatic nucleus and pineal gland. In contrast, estrogen increases the density of beta 1 receptors in the medial preoptic nucleus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559343 TI - Inhibition by catecholaminergic blockers of the neurotensin-induced enhancement of luteinizing hormone secretion in the ovariectomized estrogen-primed rat. AB - We have shown that the injection of neurotensin (NT) in the medial preoptic area significantly enhanced the estrogen-induced circadian secretion of luteinizing hormone (LH) in the ovariectomized rat. Unaltered timing of the LH secretion, despite the increased magnitude of its secretion, after the peptide injection has suggested a certain relationship between NT and other neurotransmitter systems which are involved in regulating LH secretion. The present study was undertaken to examine whether the activity of brain catecholaminergic systems are necessary for the manifestation of the peptide effect. The medial preoptic injection of NT elevated the peak value of the estrogen-induced LH secretion in vehicle-treated ovariectomized rats. Pretreatment of the animal with either alpha-methyl-p tyrosine, pimozide or phenoxybenzamine delayed the timing or reduced the peak value of the LH surge. In these drug-treated animals, the preoptic NT injection failed to induce a change in serum LH levels. It is concluded that dopaminergic and alpha-adrenergic systems are required to be intact for the manifestation of the peptide effect on LH secretion. PMID- 2559342 TI - Hyperprolactinemia decreases naloxone binding in the arcuate nucleus of ovariectomized rats. AB - Hyperprolactinemia suppresses endogenous prolactin (PRL) secretion and inhibits LH release in ovariectomized rats. Opiate peptides appear to mediate the suppressive effects of hyperprolactinemia on both endogenous PRL and LH secretion. A mechanism by which hyperprolactinemia may change the ability of opiates to influence PRL and LH is by altering the density of opiate receptors. We, therefore, examined the effect of hyperprolactinemia on the density of naloxone binding sites in hypothalamic regions that are important in the regulation of PRL and LH secretion. Female rats, ovariectomized for 4 days, were treated with ovine prolactin (oPRL) every 8 h for 2 days, and naloxone binding sites were measured using autoradiographic procedures. oPRL treatment suppressed the concentration of naloxone binding sites throughout the arcuate nucleus but had no effect in the median eminence, suprachiasmatic nucleus, and medial preoptic nucleus. There is evidence that the tuberoinfundibular dopaminergic neurons of the arcuate nucleus are directly influenced through opiate receptors. We propose that the observed decrease in the density of opiate receptors may occur on dopaminergic neurons. This theory provides an explanation for a mechanism for the suppression of endogenous PRL and LH by hyperprolactinemia: a decrease in opiate receptors will decrease opiate suppression of dopamine neurons allowing dopamine activity to increase. Increase in dopamine release are known to decrease PRL and LH secretion in ovariectomized rats. Alternatively, decreased naloxone binding may result from homologous down-regulation of receptors due to increased opiate activity. If opiate activity increases, it may directly inhibit LHRH neurons and may suppress the activity of inhibitory neurons leading to increased dopamine activity. PMID- 2559344 TI - Arachidonic acid metabolites modulate rat hypothalamic corticotropin-releasing hormone secretion in vitro. AB - Arachidonic acid metabolites have been shown to modulate the secretion of various hormones, including luteinizing hormone, growth hormone and adrenocorticotropin. In this paper we describe the effect of a series of eicosanoids on hypothalamic secretion of corticotropin-releasing hormone (CRH) in vitro. Explanted rat hypothalami in culture were exposed to prostaglandins (PG) F2 alpha or E2, thromboxane (TX) B2, the TXA2 receptor agonist U-49,619 and leukotrienes (LT) B4, C4 and D4 at concentrations ranging from 10(-15) to 10(-5) M. PGE2, LTD4 and TXB2 did not alter hypothalamic CRH secretion. On the other hand, the remaining eicosanoids tested induced a significant increase of hypothalamic CRH secretion (p less than 0.05). The concentration of 10(-11) M dexamethasone inhibited the effect of stimulatory eicosanoids on CRH secretion. The CRH response to U-49,619 was completely prevented by the TXA2 receptor antagonist SQ-29,548. The latter also inhibited serotonin (5-HT)-, acetylcholine (ACh)- and PGF2 alpha-induced CRH release. Indomethacin was capable of blocking the secretion of CRH induced by 5 HT and ACh. In addition, PGE2 inhibited the increase of CRH secretion induced by PGF2 alpha, 5-HT and ACh. These findings suggest that eicosanoids may be involved in the regulation of hypothalamic CRH secretion, either as autocrine/paracrine or as endocrine factors. PMID- 2559346 TI - Effect of barbiturates on polyphosphoinositide biosynthesis and protein kinase C activity in synaptosomes. AB - Previously, it has been found that phenobarbital inhibited protein kinase C (PKC) and the enzymes of the metabolism of polyphosphoinositide, especially phosphatidylinositol 4-phosphate (PIP) kinase (PIP-kinase). As a continuation of these studies, a number of barbiturates (barbituric acid, barbital, butabarbital, pentobarbital, amobarbital, phenobarbital, secobarbital and hexobarbital) were tested for inhibition of these enzymes and also of phosphatidylinositol (PI) kinase (PI-kinase), in a synaptosomal preparation at pH 7.8 from the brain of rat. All compounds, except barbituric acid (and Na-barbital for PI-kinase) inhibited the three kinases. However, PKC was approximately 3-5 fold more sensitive to inhibition by the drugs (measured by Ki values) than PIP-kinase, which was 2- to 4-fold more sensitive than PI-kinase. The inhibitory potency of the drugs increased with their lipophilicity, although to a lesser degree than expected from the differences in partition coefficients; the largest deviation from a positive correlation (i.e. hexobarbital) may be the result of the blockade of an imide (-NH) group at one position of the barbituric ring. Concentrations of drugs (after correction for the greater than normal ionization (pH 7.8) of the drugs in the assays) necessary for half-maximal inhibition were well within, or smaller than, those reported necessary for in vitro blocking of nerves. The possibility, therefore, exists that the physiological effects of the barbiturates are, in part, the result of an inhibition of protein kinase C and PIP-kinase. PMID- 2559348 TI - A study of the interaction of the alkylating agent, NIH10236, with opioid receptors in vitro and in vivo. AB - The series of experiments reported in this paper examined the spectrum of subtypes of opioid receptors alkylated in vitro by N-cyclopropylmethyl-7 alpha methylfumaramido-6,14- endoethenotetrahydronororipavine (NIH10236) and four optical isomers of the methylfumaramidophenethyl derivatives of 3-methylfentanyl. Pretreatment of membranes with NIH10236 resulted in a wash-resistant inhibition of the binding of [3H]6 beta-fluoro-6-desoxyoxymorphone (mu binding sites), the binding of [3H][D-ala2,D-leu5]-enkephalin (both the higher and lower affinity delta binding sites) and was without effect on kappa binding sites labelled with [3H]bremazocine. All four potential alkylating derivatives of 3-methylfentanyl were inactive. Pretreatment of membranes with 1 microM of the reversible ligands, (+)-cis-3-methylfentanyl, but not its enantiomer, inhibited the binding of [3H]6 beta-fluoro-6-desoxyoxymorphone and the binding of [3H][D-ala2,D-leu5]enkephalin to the lower affinity binding sites by over 90%. This phenomenon is termed "pseudo-irreversible inhibition." Incubation of pretreated membranes for 60 min at 37 degrees C, in the presence of 200 mM NaCl and 50 microM GppNHp, only partially reversed the masking of opioid receptors by (+)-cis-3-methylfentanyl. For in vivo experiments, membranes were prepared 18-24 hr after the intracerebroventricular administration of 80 and 50 micrograms of NIH10236. This resulted in decreased labelling of mu binding sites, lower affinity [3H][D-ala2,D leu5]enkephalin binding sites, as well as kappa binding sites, labelled by [3H]U69,593 and [3H]bremazocine. There was no apparent alteration in the higher affinity [3H][D-ala2,D-leu5]enkephalin binding site.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559345 TI - Intrapituitary cell-cell communication regulates ACTH secretion. AB - Exposure of rat anterior pituitary cells in culture to a cytotoxin specific for CRF-target cells is followed by increased basal ACTH secretion from the remaining cells three days after the last exposure to the cytotoxin. When CRF-target depleted populations are co-cultured with unexposed pituitary cells ACTH secretion is more than double that predicted in the absence of cell-to-cell interaction. These data provide evidence for an intrapituitary cell-to-cell signalling system modulating ACTH secretion. PMID- 2559347 TI - Lack of significant changes in mu, delta opioid binding sites and neutral endopeptidase EC 3.4.24.11 in the brain and spinal cord of arthritic rats. AB - The possible changes in neutral endopeptidase EC 3.4.24.11 ("enkephalinase", NEP), mu and delta opioid binding sites, were investigated using in vitro quantitative radioautography in various regions of the central nervous system of the Freund's adjuvant-induced arthritic rat, a model of chronic pain. Enkephalinase was labeled by a specific tritiated inhibitor, [3H]N-[(2RS)-3 hydroxyaminocarbonyl-2-benzyl-1-oxopropyl]glycine ([3H]HACBO-Gly), while mu and delta opioid binding sites were selectively labelled with [3H]Tyr-D-Ala-Gly (Me)Phe-Gly-ol ([3H]DAGO) and [3H]Tyr-D-Thr-Gly-Phe-Leu-Thr ([3H]DTLFT), respectively. As compared to controls, no significant modifications were found in NEP, mu or delta binding sites at both supraspinal and spinal levels of arthritic rats. These results suggest that the enhanced efficiency of exogenous opioids or endogenous enkephalins, reported to occur in this model of chronic inflammatory pain, are not directly related to changes in mu and delta opioid binding sites or steady state levels of NEP. PMID- 2559349 TI - The 5-HT3 receptor ligand, [3H]BRL 43694, binds to presynaptic sites in the nucleus tractus solitarius of the rat. AB - This study has employed receptor autoradiography to localise the distribution of binding sites for the 5-HT3 receptor ligand [3H]BRL 43694 in sections of the brain of the rat (using a concentration of 10 nM [3H]BRL 43694 with 100 microM GR38032F to define non-specific binding). The highest density of binding sites for [3H]BRL 43694 was observed in the nucleus tractus solitarius and amounted to 652 fmol/mg tissue. The binding of [3H]BRL 43694 was also examined in sections prepared 10 days after unilateral nodose ganglionectomy, in an attempt to determine the neuronal location of these binding sites. Denervation reduced the binding of [3H]BRL 43694 by around 50% in the ipsilateral side of the nucleus tractus solitarius, relative to the contralateral side. This would indicate that the 5-HT3 binding sites may have a presynaptic location on vagal afferent terminals. PMID- 2559350 TI - A pharmacological study of dopaminergic receptors in planaria. AB - The dopaminergic receptors of planaria have been studied with pharmacological and biochemical criteria. Dopamine D1 selective agonists (CY 208243 (10 micrograms/ml) and SKF 38393 (10 micrograms/ml] induced in planaria typical screw like hyperkinesias, that were inhibited by a D1 antagonist (SCH 23390 (10 micrograms/ml], but not by a D2 antagonist (sulpiride (1000 micrograms/ml]. Dopamine D2 selective agonists (PHNO (5 micrograms/ml), lisuride (5 micrograms/ml] on the contrary induced a typical "C" like curling, that was inhibited by pretreatment with D2 selective blocking agents, but not by D1 selective blocking agents. With agonists with a D1/D2 mixed action (apomorphine 60 micrograms/ml) or with amphetamine (100 micrograms/ml), the D1 type movements appeared to be more evident. Dopamine D1-selective agonists, mixed action agonists or D2-selective agonists, all induced a significant increase in levels of cAMP, that was prevented by pretreatment with the specific DA blocking agent. PMID- 2559352 TI - Evidence for the involvement of the central adrenergic system in interleukin 1 induced adrenocortical response. AB - The role of central catecholamines in the mediation of adrenocortical activation, induced by interleukin 1 (IL-1), was investigated by measuring ACTH and corticosterone in serum. Adult male rats were injected with either vehicle or the neurotoxin 6-hydroxydopamine (6-OHDA) into the lateral ventricle or the ventral noradrenergic ascending bundle. In vehicle-injected rats, 2 U of IL-1, injected intraventricularly, produced a 5- and 15-fold increase in ACTH and CS, respectively, in serum, 120 min after the injection of IL-1. In contrast, 6-OHDA, injected either intraventricularly or into the ventral noradrenergic ascending bundle, abolished the response to an intracerebral injection of IL-1. In addition, in rats pretreated with the alpha 1-adrenergic antagonist, prazosin, IL 1 failed to activate the adrenocortical axis. In other rats pretreated with the beta-adrenergic antagonist, propranolol, the adrenocortical response did not significantly differ from that of vehicle-pretreated rats. These results suggest that central adrenergic transmission, originating at the ventral noradrenergic ascending bundle and acting through alpha 1-adrenergic receptors, is involved in the adrenocortical response to IL-1. PMID- 2559351 TI - Involvement of alpha 1-adrenoceptors in the depolarizing but not the hyperpolarizing responses of motorneurones in the neonate rat to noradrenaline. AB - Superfusion of the hemisected lumbar spinal cord in the neonate rat with solutions containing 10(-6) to 10(-3) M noradrenaline (NA), elicited graded depolarizations recorded from ventral roots, with a mean EC50 value of 15.1 +/- 1.5 microM (mean +/- SEM, n = 37). Repeated concentration-response curves to NA could be determined from the same preparation. Adrenaline had a similar depolarizing action (EC50 9.9 +/- 1.7 microM, n = 11). Blockade of neuronal uptake of NA by desipramine (2 x 10(-6) M) caused some potentiation of submaximal responses to NA and shifted the EC50 to 6.0 +/- 1.7 microM (mean +/- SEM, n = 14). The depolarizing response to NA was unaffected by DL-propranolol (10(-7) M) or yohimbine (10(-7) M). Prazosin (5 x 10(-9), 10(-8) and 10(-7) M) reduced the responses and caused a progressive rightward shift of the concentration-response curve. The onset of blockade by prazosin was slow, superfusion for at least 90 120 min being required before the blockade plateaued. Prazosin (5 x 10(-9) and 10(-8) M) caused a surmountable blockade, the apparent pA2 being 8.3 +/- 0.2 (mean +/- SEM, N = 9). Depolarizations induced by NA were also antagonised by phentolamine (10(-6) M). An initial hyperpolarizing response to NA was unmasked after exposure to prazosin in 90% of preparations and was associated with a reduction in the spontaneous activity of the motorneurones. Both the hyperpolarization and reduction in spontaneous activity were attenuated by yohimbine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559353 TI - Exploring delta-receptor function using the selective opioid antagonist naltrindole. AB - Until recently the only pharmacological probes for delta-receptors have been peptide enkephalin analogues. These suffer from a number of limitations including high cost, partial agonist effects and a propensity for neurotoxicity. A stable non-peptide antagonist, naltrindole, has recently become available. We have explored its intrinsic actions and found that it attenuated swim stress-induced antinociception, a model for endogenous delta-receptor activation. Naltrindole may therefore be a useful alternative to presently available delta-receptor antagonists. PMID- 2559354 TI - Opioid control of the in vitro release of calcitonin gene-related peptide from primary afferent fibres projecting in the rat cervical cord. AB - In vitro superfusion of slices from the dorsal half of the rat cervical enlargement allowed the measurement of spontaneous, K+ (30 mM)- and capsaicin (0.5 microM)-evoked release of calcitonin gene-related peptide-like immunoreactive material (CGRPLI). The greater part of this immunoreactive material originated in primary afferent fibres since dorsal rhizotomy from C4 to Th2 (8 days before sacrifice) resulted in a 85-90% decrease in CGRPLI release. CGRPLI outflow which persisted after dorsal rhizotomy could still be enhanced by K+-induced depolarization but was no longer sensitive to the stimulatory effect of 0.5 microM capsaicin. Both delta (DTLET, D-Pen2-D-Pen5-enkephalin) and mu (DAGO, PL 017) opioid receptor agonists reduced the K+ evoked release of CGRPLI from the dorsal half of the cervical enlargement. Morphine was also inhibitory but the selective K opioid agonist U 69593 was inactive. As expected from the involvement of delta and mu receptors, the selective opioid antagonist ICI 174864 and naloxone prevented the inhibitory effects of DTLET and DAGO, respectively. These data suggest that opioid-induced presynaptic inhibiton of CGRP-containing primary afferent fibres may be involved in the analgesic effect of intrathecally injected delta and mu opioid agonists in rats. PMID- 2559355 TI - Brain natriuretic peptide (BNP) causes endothelium-independent relaxation and elevation of cyclic GMP in rat thoracic aorta. AB - The novel neuropeptide, brain natriuretic peptide (BNP), causes concentration dependent relaxations in rat isolated arterial rings. The pD2 value of BNP in rat thoracic aorta is 8.05 +/- 0.06, almost identical to the pD2 value of atrial natriuretic peptide (the 28 amino acid peptide, rat sequence, AP-28, 8.11 +/- 0.08), indicating that BNP and ANP have the same potency in relaxing thoracic aorta. In addition, BNP is equally potent at causing relaxation in abdominal aorta and mesenteric and renal arteries. However, BNP is less potent in causing vasorelaxation in the common iliac and femoral arteries and shows no relaxant effects in caudal arteries. This pharmacological profile of BNP in different rat arteries is very similar to that of ANP. Like ANP, BNP induces a vasorelaxation that is independent of endothelium and is associated with very sustained increases in cyclic GMP, but not cyclic AMP, levels in rat thoracic aorta. The BNP-induced cyclic GMP elevation, like the vasorelaxation, is also independent of endothelium and is not blocked by methylene blue (10 microM), a soluble guanylate cyclase inhibitor. Furthermore, BNP-induced cyclic GMP elevation is independent of extracellular calcium and potentiated by the cyclic GMP-phosphodiesterase inhibitor M & B 22948. Therefore, the pharmacological characteristics of BNP in rat blood vessels are very similar to those of ANP, suggesting that BNP and ANP may act through a common receptor and post-receptor mechanism to cause vasodilation. PMID- 2559356 TI - Insulin and IGF-I receptors in neuroblastoma cells: increases in mRNA and binding produced by glyburide. AB - Insulin and IGF-I binding to neuroblastoma cells (SK-N-MC) was increased by 13% and 7% respectively following a 24hr, incubation with the sulphonylurea glyburide. This increase in binding was associated with increased steady-state levels of insulin receptor and IGF-I receptor mRNA levels. Though insulin and IGF I both stimulate glucose uptake into these cells, the increased binding following glyburide treatment was not associated with any change in glucose uptake. PMID- 2559357 TI - Effects of the selective alpha 1-adrenoceptor blocker prazosin on EEG sleep and waking stages in the rat. AB - In order to gain a better understanding of the role of the noradrenergic system in the control of the EEG sleep-waking stages, the effects of the selective alpha 1-antagonist prazosin was investigated in the rat. Oral doses of prazosin (0.1-10 mg/kg) were administered that have been shown to enter the brain. EEG sleep and waking stages were recorded either during 8 h after drug administration at 8.00 in the morning or during 48 h after drug administration at 16.00 in the afternoon. It was found that prazosin at doses of 0.1-10 mg/kg shortened quiet waking. Starting at 1 mg/kg paradoxical sleep (PS) was shortened and, most interestingly, active waking and slow wave sleep (SWS) were prolonged. PS spindles and dozing were shortened after a latency of some hours during the 48 hour experiment. However, during the 8-hour experiment PS spindles were prolonged at 0.32 and 1 mg/kg. These data suggest that in the rat alpha 1-adrenoceptor inhibition in the brain allows the occurrence of active waking and SWS and suppresses PS. PMID- 2559358 TI - Electrophysiological indices of central and peripheral nervous system function during theophylline therapy. AB - Electrophysiological parameters including the EEG, somatosensory evoked potentials (SEP), F waves, long loop reflexes and peripheral nerve conduction velocities were assessed during the 15th week of theophylline therapy, with serum levels maintained in the low therapeutic range, in young, healthy male volunteers (n = 7). Recordings were repeated 1 week after vitamin B6 supplementation and finally 6-7 weeks after the cessation of medication. Alpha amplitudes were significantly lower during theophylline therapy compared to the posttreatment baseline recording. The SEP findings failed to reveal any significant differences between the three recordings, as was the case with peripheral nerve conduction velocities and long loop reflexes. The average F wave latencies during theophylline therapy were significantly shorter, and the percentage F waves recorded was higher, compared to the baseline recordings. These findings suggest that relatively long-term theophylline therapy has stimulatory effects on aspects of electrical activity in the brain and spinal cord. These effects appeared to be unchanged after vitamin B6 supplementation for 1 week. PMID- 2559359 TI - Involvement of sulfhydryl groups in the transport of L-tyrosine and L-tryptophan across the human red cell membrane in vitro. AB - In previous papers, we reported a deficit in tyrosine (TYR) and tryptophan (TRP) transport across the erythrocyte membrane in depressed patients. To investigate further the transport mechanism of the two precursors of monoamines, we tested in healthy subjects the role played by sulfhydryl groups (SH). These groups, cysteine residues, are localized on the intrinsic domain such as the transporters of chloride or sugars. We found that all sulfhydryl reagents that inactivated the SH induced a strong inhibition of the transport of amino acid across the red cell membrane when incubated in the plasma as medium. We concluded that a relationship might exist between these neutral amino acids and D-glucose transport. PMID- 2559360 TI - Effects and after-effects of the common cold and influenza on human performance. AB - Volunteers who develop a cold following virus challenge were significantly slower on choice reaction time tasks than those with no illness. This effect was still observed after the clinical symptoms had gone. In contrast to this, influenza illnesses only impaired performance in tasks in which subjects were uncertain where the target stimulus would appear. These results demonstrate that the CNS effects of respiratory virus infections depend on the type of virus, and that performance impairments may remain even after the symptoms of a cold have gone. PMID- 2559361 TI - Potential antidepressant properties of subchronic GABA transaminase inhibitors in the forced swimming test in mice. AB - We carried out the forced swimming test in mice to investigate the antidepressant potentials of GABA transaminase (GABA-T) inhibitors including aminooxyacetic acid, ethanolamine-O-sulfate, gamma-vinyl GABA (GVG) and valproic acid (VPA). In acute experiments only GVG reduced immobility. Following chronic oral administration via drinking water containing the drugs, immobility was significantly reduced at days 5 and 10 in all of the GABA-T inhibitors examined. The tolerance of the anti-immobility effect, however, occurred at day 20. Brain GABA contents showed moderate to marked increase during the session with the exception of VPA, which did not alter GABA contents. These results suggest that subchronic GABA-T inhibitors possess antidepressant properties, though the anti immobility effect of GABA-T inhibitors did not directly correlate with the increase in brain GABA. PMID- 2559362 TI - Malignant clear cell hidradenoma of the lip. AB - An interesting variety of cutaneous tumors of the head and neck, tumors of the eccrine differentiation, are seldom recognized. The most recent reports of these tumors appear in the dermatologic literature, probably due to the majority of them being located in the trunk and the extremities. Malignant varieties of these tumors do exist and must be treated as aggressively as more common tumors of the head and neck region. Tumors with eccrine differentiation are among a large variety of cutaneous tumors that present in the head and neck. These tumors are far less common than basal cell and squamous cell carcinomas of the head and neck. Malignant clear cell hidradenomas, a variant of the more common benign clear cell hidradenoma, is one such type of tumor of the eccrine differentiation. This is a rare tumor seldom reported in sites of the head and neck. A case of a 47-year-old white male with such a tumor of the lower half of his lip is presented. This report is particularly concerned with the histopathology of the tumor, its unique etiology, and the applicability of the lip switch flap (Stein Abbe-Estlander) in the reconstruction of the surgical defect following total tumor excision. PMID- 2559363 TI - [Primary malignant neoplasms of the liver. Contribution of echography to the preoperative diagnosis]. AB - Primary malignant hepatic tumors are relatively common and the possibility of radical surgical treatment in the initial stages has modified our clinical approach therefore stressing the validity of a correct screening and follow-up of this pathology. The Authors present their clinical experience in the field enriched by the use of ultrasonography and conclude that this represents a highly effective diagnostic technique whereas it is less reliable in the determination of the nature of the lesion. PMID- 2559364 TI - [Radical surgery and loco-regional chemotherapy in the treatment of liver metastases of stomach and colorectal cancer]. AB - The results of surgical treatment of hepatic metastasis from stomach and colo rectal cancer in a series of 28 patients have been assessed. 21 cases were subjected to hepatic resection (stomach cancer: 7 cases; colorectal: 14 cases) and 7 underwent locoregional chemotherapy through the gastroduodenal artery with fully implantable catheters (stomach ca.: 1 case; colorectal ca.: 6 cases). Of patients who underwent hepatic resection, 3 died postoperatively and 4 are still alive 5 years after the operation. Of the 7 patients treated with locoregional chemotherapy, 4 can be evaluated: 2 responded to treatment, 1 stabilised and 1 progressed. Clinically noteworthy toxicity was observed in one patient only. PMID- 2559365 TI - Ganciclovir therapy of symptomatic cytomegalovirus infection in renal transplant recipients. AB - We used ganciclovir to treat 11 renal transplant recipients with symptomatic cytomegalovirus infection (seven primary), including one severe, five mild and five moderate cases. Two patients exhibited a non-mechanically ventilated pneumonitis and two others a gastrointestinal involvement. Ganciclovir was used intravenously according to a schedule which took into account renal function, for a median time of 14 days. All patients survived. Cytomegalovirus infection was cured in all patients but two: in the first an early clinical relapse required a second successful ganciclovir course; in the other graftectomy was needed to control infection. Graft was lost in an additional cured patient. Ganciclovir was well tolerated, especially with regard to haematological status. At the current follow-up of at least one month after the end of ganciclovir therapy, no further clinical relapse was observed; however, in one clinically cured patient cytomegalovirus was isolated from blood one week after ganciclovir cessation. These encouraging preliminary data suggest that ganciclovir therapy should be started as soon as cytomegalovirus infection is suspected, especially in cytomegalovirus seronegative recipients receiving a seropositive graft. PMID- 2559366 TI - Blockade of the late IPSP in rat CA1 hippocampal neurons by 2-hydroxy-saclofen. AB - The effects of the GABAB receptor antagonist 2-hydroxy-saclofen were studied using intracellular recording of synaptic potential from CA1 hippocampal neurons. 2-Hydroxy-saclofen (50-200 microM) reversibly blocked the late, GABAB receptor mediated inhibitory postsynaptic potential (IPSP) but not the early, GABAA receptor-mediated IPSP. In addition, the hyperpolarizing response to baclofen was reduced by similar concentrations of 2-hydroxy-saclofen. This suggests that 2 hydroxy-saclofen is a potent antagonist at postsynaptic GABAB receptors on hippocampal neurons. PMID- 2559367 TI - Adrenocortical response following acute neurogenic stimuli is mediated by CRF-41. AB - The present study examined whether neurogenic stimuli activate the pituitary adrenal axis via CRF-41. Adult male rats were exposed to photic, acoustic or sciatic nerve stimulation. At 4, 15, and 30 min following the onset of stress, animals were sacrificed, trunk blood collected and the median eminence removed. At 4 min following the stress onset, there was a significant decrease in CRF-41 content of the median eminence, which persisted for 30 min. Concomitant with the decrease in CRF-41 content, serum adrenocorticotropic hormone (ACTH) and corticosterone levels increased. Thus, this study demonstrates that CRF-41 released from the median eminence plays a dynamic role in mediating the ACTH and corticosterone response to neurogenic stimuli. PMID- 2559368 TI - Increased responsiveness of the cerebral cortical phosphatidylinositol system to noradrenaline and carbachol in senescent rats. AB - The responsiveness of cerebral cortical alpha 1-adrenoceptors and cholinergic muscarinic M1 receptors was assessed in young (3 months) and aged (24 months) male Sprague-Dawley rats. The measure of responsiveness was the accumulation of inositol phosphate (IP) formed in [3H]myo-inositol-preloaded cerebral cortical slices in the presence of lithium, following stimulation with various concentrations of noradrenaline (1-300 microM) and carbachol (5-1000 microM). In old rats the maximum response to noradrenaline was higher by 80%, and that to carbachol by 33%, indicating an increased responsiveness of the investigated receptors in senescence. PMID- 2559369 TI - Kainate evokes the release of endogenous glycine from striatal neurons in primary culture. AB - The actions of 56 mM KCl and excitatory amino acid (EAA) agonists on the release of endogenous glycine (Gly) from striatal neurons in primary culture was examined. During a 3 min period, 2 x 10(6) striatal neurons released 743 +/- 51 pmol of Gly. In the presence of 56 mM KCl, an additional 492 +/- 52 pmol of Gly (+66%) were released, 75% of which was dependent upon the presence of extracellular calcium. When striatal neurons were exposed to 1 mM N-methyl-D aspartate (NMDA) or quisqualate (QA), endogenous Gly released was increased by 370 +/- 71 (+50%) or 120 +/- 31 (+16%) pmol, respectively. In the presence of 1 mM kainate (KA), however, the release of endogenous Gly increased by 994 +/- 82 pmol (+135%). Interestingly, while KA (1 mM) was twice as effective as KCl (56 mM) in evoking the release of endogenous Gly, KCl was 5 times more effective than KA in evoking the release of endogenous gamma-aminobutyric acid (GABA). KA induced increases of endogenously released Gly were dose-dependent (EC50, 100 microM), saturable and not significantly reduced in the absence of extracellular calcium. The actions of KA were blocked by coincubation with 6-cyano-2,3 dihydroxy-7-nitro-quinoxaline (CNQX), a competitive antagonist at the KA receptor. These data suggest that the release of endogenous Gly from striatal neurons in primary culture is regulated principally by EAA actions at the KA receptor system. PMID- 2559370 TI - Antagonism at GABAB receptors by saclofen and related sulphonic analogues of baclofen and GABA. AB - Saclofen (the direct sulphonic analogue of baclofen) is a competitive antagonist of baclofen at GABAB receptors in guinea pig ileum and rat cortical slices (estimated pA2 = 5.3), at least twice as potent as 2-hydroxy-saclofen (pA2 = 5). A series of related sulphonic analogues also antagonised baclofen in the guinea pig ileum, including 2-hydroxy-saclofen amide (pA2 = 3.3), 3-amino-2-hydroxy-2 phenyl-propylsulphonic acid (pA2 = 3.5), 3-amino-2-(benzo-(b)-furan-2-yl) propylsulphonic acid (pA2 = 4.3), and 3-amino-2-(4-chlorophenyl)-prop-1 enesulphonic acid (pA2 = 2.5-3), but none were more active than saclofen which is the most potent specific GABAB antagonist yet found. PMID- 2559371 TI - Stimulation of the rostral ventrolateral medullary neurons increases cortical cerebral blood flow via activation of the intracerebral neural pathway. AB - In urethane-anesthetized, artificially ventilated rats, electrical or chemical (by L-glutamate) focal stimulations of the rostral ventrolateral medulla (RVLM) produced an increase in cortical cerebral blood flow (CBF). The RVLM-induced cortical vasodilative response was present in animals with spinal cords sectioned at levels of Th3-4 and with bilateral extracerebral cervical sympathetic trunks (CSTs) severed. The RVLM-induced cortical vasodilative response was totally eliminated by an alpha 2 adrenergic blocker, but not by blockers for muscarinic, nicotinic, alpha 1 and beta receptors. It was concluded that there is an intracerebral vasodilative neural pathway including an alpha 2 adrenergic receptor originating in the RVLM for regulation of cortical blood vessels. PMID- 2559372 TI - Embryonic and postnatal development of N-(1-[2-thienyl]cyclohexyl)[3H]piperidine binding sites in rat forebrain homogenates and slices. AB - The development of N-(1-[2-thienyl]-cyclohexyl)[3H]piperidine [( 3H]TCP) binding to phencyclidine (PCP) receptors in both brain homogenates and slices has been investigated in the rat. The specific binding sites for [3H]TCP in the homogenate were already detected at prenatal stages and steadily increased after birth. A similar developmental pattern was seen in the autoradiography of the [3H]TCP binding to the brain slice in which the distribution of the binding in the young is more homogeneous than that in adult. There was an increase in the Bmax without changes in the Kd of the [3H]TCP binding and there was no change in inhibition of the binding by TCP, PCP and D-(-)-2-amino-5-phosphonovalerate during postnatal maturation. These findings suggest an increase in the density with no change in the affinity of PCP receptors and the absence of a change in the interaction between the PCP and N-methyl-D-aspartate receptors in the developing rat forebrain. PMID- 2559373 TI - Activation of the classical complement pathway in brain tissue of Alzheimer patients. AB - Positive immunohistochemical staining of Alzheimer brain tissue was obtained with antibodies to proteins associated with classical, but not the alternative, complement pathway. Clq, C3d, C4d are fractions of complement proteins that bind to tissue when the classical complement pathway is activated. Antibodies to these fractions stained senile plaques, dystrophic neurites and some neurofibrillary tangles. C5b-9 is the membrane attack complex which promotes cell lysis when assembled on the plasma membrane. An antibody to a neoantigenic site on this complex stained dystrophic neurites and many neurofibrillary tangles, but not extracellular amyloid. Properdin and fraction Bb of factor B, two proteins that bind to tissue when the alternative complement pathway is activated, were not detected immunohistochemically. PMID- 2559374 TI - Elevation of intracellular calcium content in area CA1 of hippocampus is not directly correlated with the development of long-term potentiation. AB - Electron microscopic localization of calcium-containing mitochondria in stratum radiatum of CAl of hippocampal slices was performed after (1) low-frequency stimulation, (2) high-frequency stimulation, and (3) blocking N-methyl-D aspartate (NMDA) receptors during high-frequency stimulation. Dendritic mitochondria containing Ca deposits were found in a narrow band of stratum radiatum 280-350 microns distant from stratum pyramidale. Axonal mitochondria containing Ca deposits were evenly distributed in stratum radiatum. The total number of calcium containing-mitochondria was highest in long-term potentiated slices, and less in slices treated with APV; the lowest values were obtained with low-frequency stimulation. PMID- 2559375 TI - Hydrogen sulfide in combination with taurine or cysteic acid reversibly abolishes sodium currents in neuroblastoma cells. AB - Patch clamp studies of neuroblastoma cells have shown that in the presence of sodium hydrogen sulfide (NaHS; the in vitro precursor of H2S), addition of the sulfonated amino acids, taurine or cysteic acid resulted in reversible abolition of the inward sodium currents. This effect could also be demonstrated by preincubating cells for 3-20 min with 5-10 mM NaHS followed by replacement of the solution with taurine or cysteic acid in sulfide-free saline. Neither NaHS, taurine nor cysteic acid alone had any effect. The sulfhydryl reagents, beta mercaptoethanol and dithiothreitol, were also found to abolish reversibly the sodium currents. As the effects of the above treatments were nearly identical, the synergistic action of NaHS with taurine or cysteic acid may result from reduction of the disulfide bonds between subunits comprising the sodium channel. The responses to NaHS and taurine, a putative neurotransmitter/neuromodulator, suggest that reductions in sodium channel function may be the mechanism(s) responsible for loss of central respiratory drive during H2S poisoning. PMID- 2559376 TI - The effects of cyanide on neural and synaptic function in hippocampal slices. AB - Transverse slices from guinea pig hippocampi were exposed to micromolar concentrations of sodium cyanide while neural and synaptic function were monitored in the CA1 region. Cyanide concentrations between 10 and 200 microM rapidly depressed synaptic transmission between Schaffer collateral-commissural fibers and CA1 pyramidal cells. Analysis of input/output curves revealed that the suppression had two components, a decrease in EPSP generation and an increase in action potential threshold. Direct electrical excitability of axons was not affected. At concentrations to 500 microM, cyanide had no effect on antidromic activation of pyramidal cells. At 1000 microM, cyanide caused a moderate depression of the antidromic response in one slice while having no effect in one other. In some experiments, postsynaptic responses in the gyrus dentatus (GD), evoked by perforant path stimulation, were recorded simultaneously with CA1 responses during cyanide application. GD was found to be less sensitive to cyanide than CA1. All cyanide effects reversed rapidly and completely upon washout. These findings suggest that cyanide has a direct effect on neurons not mediated by its inhibition of metabolism. PMID- 2559377 TI - Pharmacological properties of quisqualate- and kainate-preferring glutamate receptors induced in Xenopus oocytes by rat and chick brain mRNA. AB - Quisqualate- and kainate-preferring glutamate receptors were studied in Xenopus oocytes injected with rat and chick brain mRNA. All ligands effective at ionotropic quisqualate sites, agonists as well as antagonists, were found to inhibit kainate responses. Ionotropic quisqualate and kainate responses were distinguished by gamma-L-glutamylglycine but not by its D-isomer. No difference was detected between responses induced by rat brain mRNA and chick brain mRNA, suggesting that the properties of non-N-methyl-D-aspartate-type glutamate receptors of rat and chick brains are essentially the same. PMID- 2559378 TI - Long-term desensitization of quisqualate-specific glutamate receptors in Purkinje cells investigated with wedge recording from rat cerebellar slices. AB - Responses of Purkinje cells to quisqualate and aspartate were examined in rat cerebellar slices with a 'wedge' recording technique. Conditioning with bath application of 100 microM quisqualate for 1 or 4 min frequently induced reduction of Purkinje cell responses to testing bath application of 100 microM quisqualate. Test responses obtained at 5-30 min intervals were 79% after 1 min conditioning and 64% after 4 min conditioning, relative to conditioning responses. Since there was no correlation between the degree of reduction and the conditioning-testing interval, the effect of conditioning appeared to be sustained over 30 min. No such desensitization was observed with 3 or 5 mM aspartate. The presently demonstrated desensitization of quisqualate-specific glutamate receptors could represent a central mechanism of the 'long-term depression' type of synaptic plasticity in Purkinje cells. PMID- 2559379 TI - What is the optimal imaging time for 99Tcm-(V)-DMSA planar scintigraphy in the detection of squamous carcinoma? A comparative study in humans and in an animal tumour model. AB - 99Tcm-(V)-DMSA is a new tumour imaging agent which has been used to image squamous cell carcinoma (SCC) of the head and neck. There have been, however, no studies to date evaluating its optimal imaging time for SCC. Seven patients were studied (six SCC; one nontumour) and seven rabbits (six with SCC, (17 tumours); one nontumour). For the human qualitative studies there was a 67% sensitivity at 2, 4 and 6 h with image quality being optimum at 4 h. Maximum quantitative uptake occurred between 2 and 4 h. For the rabbit qualitative studies the optimum imaging time was 4 h (92% sensitivity, 100% specificity) and maximum quantitative uptake occurred at between 1.5 and 5 h. Taking into account the human and rabbit qualitative and quantitative studies combined with the pharmacokinetics and biodistribution of 99Tcm-(V)-DMSA, the optimum imaging time of 99Tcm-(V)-DMSA in humans with SCC was between 2 and 4 h. PMID- 2559381 TI - Imaging of small cell carcinoma using 131I-labelled antibodies to vasopressin associated human neurophysin (VP-HNP) AB - Preliminary studies are reported on the use of 131I-labelled antibodies against vasopressin associated human neurophysin to image tumors in patients with small cell carcinoma of the lung (SCCL). The rabbit polyclonal antibodies used in these studies were affinity purified on columns of neurophysin-Sepharose. Patients were pre-screened for the presence of neurophysin producing tumors by plasma RIA. Six patients who screened positive received approximately 1 mCi/70 kg body weight of radioiodinated antibody preparation, and scintigraphy was subsequently performed at 24 h, 48 h, and 72 h using a subtraction technique based on simultaneous imaging with radiopharmaceutical agents labelled with technetium-99m. Tumor was clearly demonstrated at 72 h in all five patients who had measurable lesions at the time of study, while no positive image was noted for one patient in complete clinical remission. Since approximately 70% of all SCCL patients have neurophysin producing tumors, our findings suggest that neurophysin antibodies may be effectively used to scan tumors in a majority of patients with SCCL. PMID- 2559380 TI - 99Tcm-MAG3 for quantitation of differential renal function. AB - We studied two different methods for the evaluation of differential renal function in a group of 100 patients with various kidney disorders whose effective renal plasma flow (ERPF) had been calculated previously by single 125I orthoiodohippurate (OIH) injection and multiple blood sampling. Patients were divided into three groups according to their ERPF:ERPF is greater than or equal to 250 ml min-1; ERPF less than or equal to 100 ml min-1; and ERPF greater than 100 ml min-1 and less than 250 ml min-1. The two methods used to assess differential renal function were: first, relative 99Tcm-dimercaptosuccinic acid (DMSA) uptake calculated by normalized background and attenuation corrected cumulative counts in each kidney 24 h p.i.; and second, relative 99Tcm mercaptoacetyl-triglycine (MAG3) uptake within 1 and 2 min p.i. calculated by normalized background and attenuation corrected counts on each renal area. The results obtained with each method correlated strongly with high significance (p less than 0.0001). In the right kidney, mean values obtained with 99Tcm-MAG3 tend to be higher than mean values obtained with 99Tcm-DMSA. PMID- 2559382 TI - The pyramidal lobe: a scintigraphic assessment. AB - The thyroid scintigraphs of 207 patients were examined with a view to investigate the anatomical origin of the pyramidal lobe (PL) and in order to determine the incidence of its scintigraphic detection. The PL was visualized in 41% of the cases studied with a greater incidence among the females as compared to the males. It was established that the likelihood of the PL arising from the isthmus of the thyroid or its lobes was about the same. Further, it was observed that the isthmal PL was usually located in the mid-line rather than laterally, in which case it was more often deviated to the left than to the right. The scintigraphic visualization of the PL was seen to increase with the size of the gland but in large goitres it was difficult to detect. It evolves that although at times, there might be a discrepancy between the apparent and the actual location of the PL due to the presence of overlying tissue, in a majority of cases, the scintigraphic appearances do in fact reflect the true picture. PMID- 2559383 TI - Primary mucinous sweat gland carcinoma of the eyelid. AB - A 76-year-old woman had a slowly enlarging lesion of the right lower eyelid. After obtaining incisional biopsy results consistent with adenocarcinoma, we removed the tumor and made a final diagnosis of primary mucinous adenocarcinoma of the sweat gland. The defect was repaired using ipsilateral upper lid transpositional tarso-conjunctival and skin-muscle flaps. No evidence of recurrence or metastasis was noted 42 months postoperatively. Consideration of such tumors is clinically important, since they have full metastatic potential. We therefore suggest consideration of sweat gland primary mucinous adenocarcinoma in the differential diagnosis of red or violaceous eyelid masses. PMID- 2559384 TI - Evaluation of immunologic tests for the detection of ocular herpes simplex virus. AB - Four immunologic tests, Herpchek (HC), latex agglutination (AGG), enzyme immunofiltration (IF), and 1 hour enzyme-linked immunoassay (1EIA), were evaluated for detecting herpes simplex virus (HSV) from ocular specimens. Compared with the standard of HSV-positive cell cultures, 24 (65%) of 37 positive HC tests and 22 (59%) of 37 positive IF tests were significantly more sensitive (P less than 0.0005) than the AGG (3%, 1/37) and 1EIA (26%, 1/34) tests. The HC and IF tests were 100% specific, and easy to interpret. In addition, the clinical diagnosis of HSV infection after a thorough ophthalmologic examination was as sensitive (59%, 22/37) (P = 0.4) as any of the immunologic tests. In conclusion, for definitive diagnosis of HSV, the HC test seems more suitable for a central laboratory that handles a large number of viral specimens, and processes HC negative tests with follow-up cell culture isolation. The enzyme immunofiltration test seems more suitable for a low-volume viral laboratory which confirms all IF test samples with follow-up cell culture isolation. PMID- 2559385 TI - Adenoid cystic carcinoma of the lacrimal sac. AB - Adenoid cystic carcinoma is the most common epithelial malignancy of the lacrimal gland and minor salivary glands. Its occurrence in the lacrimal drainage system is extremely rare. A 57-year-old woman presented with epiphora and a mass in the lacrimal sac, which proved to be an adenoid cystic carcinoma. Histologically, it appeared to arise from the wall of the lacrimal sac, and both the lacrimal gland and maxillary antrum were uninvolved by tumor. Despite aggressive surgical intervention, recurrence and metastasis developed. Adenoid cystic carcinoma must be considered in the differential diagnosis of epiphora. PMID- 2559386 TI - Orbital nonchromaffin paraganglioma. A case report and review of the literature. AB - Nonchromaffin paraganglioma (NCP), also called glomus body tumor or chemodectoma, is rarely found in the orbit. The behavior of orbital nonchromaffin paraganglioma may potentially be more aggressive than in other head and neck locations. Diagnosis depends on electron microscopic demonstration of membrane-bound neurosecretory granules. Results of histopathologic study show a well circumscribed lesion without a true capsule with alveolar or organoid arrangements of epithelioid cells within a reticulin framework with thin-walled blood vessels. Cells are polygonal with round or oval nuclei containing rare mitotic figures and pale-staining cytoplasm. Differential diagnosis includes alveolar soft-part sarcoma, alveolar rhabdomyosarcoma, neuroblastoma, carcinoid, and granular cell tumor. Of 29 previously reported cases of orbital NCP, 16 have been reclassified as alveolar soft-part sarcoma. The authors report a patient with an electron microscopically established orbital NCP, with the history of a contralateral glomus jugulare tumor irradiated 14 years previously. PMID- 2559387 TI - Collagen/hydroxylapatite implant for augmenting deficient alveolar ridges: a 24 month clinical and histologic summary. AB - A new alveolar ridge augmentation material composed of purified fibrillar collagen (PFC) and particulate hydroxylapatite (HA) (Alveoform Biograft, Collagen Corp., Palo Alto, Calif.) was evaluated in the treatment of 77 edentulous patients in a multicenter study. All patients had mandibular augmentation; 22 also had maxillary implant augmentation. Mean ridge height was 15.4 mm before surgery. Twenty-four-month data showed a mean ridge augmentation of 4.1 mm. Prosthodontists rated the ridge firmness "good" to "excellent" in 96.6% of patients after 2 years. All adverse effects with the exception of some residual paresthesia/dysesthesia resolved spontaneously within 24 months. Bone-graft interface samples were examined histologically in five patients 1 year after mandibular ridge augmentation. No evidence of PFC was found, and the HA particles were surrounded by dense, fibrous, host-connective tissue or trabeculae of woven and lamellar bone. Assessment of PFC/HA-augmented ridges at 24 months showed clinically and histologically significant results. PMID- 2559388 TI - The combined use of endosseous dental implants and collagen/hydroxylapatite augmentation procedures for reconstruction/augmentation of the edentulous and atrophic mandible: a preliminary report. AB - The combination of a relatively new alveolar ridge augmentation material composed of purified fibrillar bovine collagen (PFC) and particulate hydroxylapatite (HA) (Alveoform Biograft Collagen Corp., Palo Alto, Calif.), along with endosseous root form dental implants, was simultaneously placed by a combined flap and tunneling technique to reconstruct and augment endentulous and atrophic mandibles. Prosthodontic and patient surveys reported great satisfaction with overdenture stability, retention, comfort, esthetics, speech, and function. Six reported cases showed that clinically significant alveolar ridge augmentation and form were achieved and maintained with the added prosthetic support and improved stability and retention produced by the combination of procedures. Histologic study of core specimens obtained through the ridge augmentation graft material provided evidence of supracortical bone ingrowth into the graft matrix. PMID- 2559389 TI - A comparative analysis of monoaminergic involvement in the spinal antinociceptive action of DAMPGO and DPDPE. AB - The antinociceptive properties of intrathecally (i.t.) administered [D-Ala2, N methyl-Phe4, Gly5-ol]enkephalin (DAMPGO) and [D-Pen2, D-Pen5]enkephalin (DPDPE), selective opioid agonists for mu (mu) and delta (delta) sites, respectively, were compared in rats. DAMPGO and DPDPE elevated tail-flick latency (TFL) in a dose dependent manner, and the spinal antinociceptive actions of both drugs were reversed by the opiate antagonist naloxone. These findings suggest that both DAMPGO and DPDPE interact with spinal opiate receptors to elevate TFL. Another set of experiments was done to determine the involvement of local spinal serotonin (5-HT) or norepinephrine (NE) in DAMPGO and DPDPE-induced spinal analgesia. Both the alpha 1 noradrenergic receptor antagonist WB-4101 and the alpha 2 blocker yohimbine failed to alter the antinociceptive actions of DAMPGO and DPDPE. Similarly, the 5-HT receptor antagonists pindolol, ritanserin and ICS 205-930 (selective for 5-HT1, 5-HT2 and 5-HT3 sites, respectively) failed to inhibit opioid-induced spinal analgesia. Thus, while DAMPGO and DPDPE produce antinociception via an interaction with spinal opioid receptors, apparently neither drug activates endogenous monoaminergic systems. PMID- 2559390 TI - [The use of siliceous immunosorbents in hemosorption]. AB - Experiments were conducted on rabbits and monkeys for approbation of immunosorbents by covalent immobilization of human serum albumin or antibodies to human apoprotein B on the surface of silo-chromium and subsequent incubation with 1% albumin solution. Extracorporeal immunosorption in immunized rabbits led to reduction of the level of specific antibodies by 60% and the number of blood platelets by 10-30%. The initial antibody level was restored in 5-10 days. Study of immunosorbents on monkeys showed a satisfactory compatibility of the sorbent with blood, its ability for regeneration, and the possibility of its repeated use. The specific capacity was 66% of the capacity according to human apoprotein B. PMID- 2559391 TI - [Neurohormones and alcoholism]. PMID- 2559392 TI - [Radiation-induced esophageal stricture in patients with non-small cell lung cancer treated with chemoradiotherapy]. AB - Five out of 165 cases (3.0%) which were treated for non-small cell lung cancer with radiotherapy (98 cases were treated with chemoradiotherapy, and the other 67 cases, radiotherapy alone) developed esophageal stricture. Their clinical courses, the relationship among radiation dosage, combination with chemotherapy, the length of the irradiated esophagus, and the occurrence of esophageal stricture were reviewed. One of the 5 cases was a case with lung cancer in Bloom's syndrome, which developed an esophageal stricture after receiving only 30.6 Gy (the TDF value was 46.2) to the esophagus. This case suggests the possibility that a patient with Bloom's syndrome is more radiosensitive than normal controls. The other 4 cases were treated with combined chemoradiotherapy. One of the 4 cases was treated with concomitant use of bleomycin (BLM), while the TDF value was not more than 100 (75.4). The concomitant sue of BLM was almost certainly the cause of the esophageal stricture. The other 3 cases were treated with chemoradiotherapy, the TDF values of which were more than 100 (108.7, 112.5, and 129.3). The chemotherapy combined with radiotherapy and the overdosage were considered to be the cause of the esophageal stricture in these 3 cases. These data suggest that in Bloom's syndrome, radiotherapy should be performed carefully and that BLM should not be used simultaneously with irradiation to the esophagus. It is also believed that a radiation dose over 100 in TDF value to the esophagus should be discouraged when chemotherapy is combined. PMID- 2559393 TI - Prevalence of the sickle cell trait among students in a physical education college in Cote-d'Ivoire. AB - The prevalence of sickle cell trait (SCT), was studied in 263 students enrolled in a physical education college in Cote-d'Ivoire. Most of the students were athletes of average training level, but some competed at an international level. The prevalence of SCT (hemoglobin AS), observed in the student population (13.7%) was not significantly different from that of the general population (12%) of Cote d'Ivoire and did not differ between sexes. The percentage of international level athletes was similar in SCT (25%) and control (hemoglobin AA) groups (25.7%). Particular activities practised by SCT international level athletes were individual track events (sprint race, 400 m race and long and high jump) as well as team sports (football and handball), indicating the excellent aptitude for aerobic and anaerobic metabolic activities. As none of the international athletes engaged in endurance activities, no conclusions can be drawn regarding either the aptitude of SCT subjects for intense and prolonged exertion or a possible limitation of oxygen transport produced by their hemoglobinopathy. PMID- 2559394 TI - Angiotensin-converting enzyme inhibitors: mechanistic controversies. AB - Many studies have investigated the mechanisms responsible for the therapeutic effects of the angiotensin converting enzyme inhibitors. Initially, the hemodynamic changes that occur with these agents were attributed solely to the inhibition of the renin-angiotensin-aldosterone system in plasma. Further research suggested other mechanisms were operable as a relationship was not always evident between hemodynamic changes and inhibition of the plasma renin angiotensin-aldosterone system. A relationship between the pharmacodynamics of these agents and the inhibition of vascular and tissue renin-angiotensin systems, however, has been observed. Mechanisms less likely to contribute to the actions of the angiotensin converting enzyme inhibitors are increases in bradykinin and prostaglandin concentrations, or inhibition in the renin-angiotensin system within the central nervous system. Ancillary cardiovascular effects of angiotensin converting enzyme inhibitors offer possible new therapeutic gains. An understanding of these mechanistic controversies and newly-defined cardiovascular actions of angiotensin converting enzyme inhibitors are important to clinicians using these agents. PMID- 2559396 TI - What's new in human herpesvirus-6? Clinical immunopathology of the HHV-6 infection. AB - Human herpesvirus-6 (HHV-6), formerly known as human B-lymphotropic virus (HBLV), was first isolated in 1986 from patients with lymphoproliferative disorders and AIDS. Antibody prevalence against HHV-6 varies between about 60-80% indicating a widespread latent infection. Although HHV-6 infects in vivo primarily T lymphocytes, it is associated with similar diseases as in infection with Epstein Barr virus (EBV), a clearly B-lymphotropic virus. Reactivation of latent HHV-6 infection in patients with subnormal host defense may cause persistent active infection with so-called postinfectious chronic fatigue syndrome (PICFS) or may contribute to other pathologies such as immune deficiency itself, autoimmune disorders or progressive lymphoproliferation. Coinfection of CD4 cells by HHV-6 and human immunodeficiency virus (HIV 1) in AIDS patients can aggravate HIV induced acquired immune deficiency. These characteristics of the only recently detected new virus justify further intense investigation. PMID- 2559395 TI - Erythema induced by quartz-halogen sources. AB - The erythemal effect of 100-W quartz-halogen sources, previously predicted from radiation measurements, was directly measured on humans at a distance of 10 cm. Minimal erythema was produced in 15 min for skin type I. These data show that normal use of a desktop lamp should not usually lead to erythema in one day, but that long-term effects cannot be ignored, because for lifetime use at work, the relative risk for squamous cell carcinoma on the back of the hands is 3.4. PMID- 2559397 TI - The effect of prolonged ethanol administration on central alpha 2-adrenoceptors sensitivity. AB - The response of an endogenous inhibitor of protein kinases (type II inhibitor) to clonidine was used as an index of sensitivity of central alpha 2-adrenoceptors. Low doses of clonidine (20-50 micrograms/kg) induced an increase in type II inhibitor activity in the nucleus accumbens, hippocampus and in the anterior and posterior hypothalamus by stimulating presynaptic alpha 2-adrenoceptors. Stimulation of postsynaptic alpha 2-adrenoceptors by high doses of clonidine 0.5 1.0 mg/kg resulted in a dose-dependent decrease in type II inhibitor activity. Prolonged treatment with ethanol (5 g/kg/day po for 21 days) greatly reduced the action of high doses of clonidine in all the examined brain areas, suggesting subsensitivity of postsynaptic alpha 2-adrenoceptors lasting for at least 48 h after the last ethanol administration. A single dose of ethanol induced a short lasting subsensitivity of postsynaptic alpha 2-adrenoceptors in the anterior hypothalamus. 12 h after administration of alcohol the response of type II inhibitor to high doses of clonidine in this brain area was the same as in untreated rats. PMID- 2559398 TI - Rapid diagnosis of common sexually transmitted diseases in adolescents: a review. AB - Sexually transmitted diseases (STDs) represent a significant health problem for adolescents in the United States. As a result, clinicians from a number of disciplines, among them pediatrics and dermatology, may be called on to evaluate teenage patients with these infections. Traditionally, the diagnosis of many STDs has relied on isolation of the causative organisms in culture. While such procedures typically offer optimal diagnostic accuracy, they are often time consuming and expensive, and may not be widely available. In an attempt to remedy this, culture-independent techniques have been developed to permit more rapid and economical diagnosis of these important pathogens. The strengths and limitations of these methods must be understood, however, for the tests to be used effectively. PMID- 2559399 TI - Pediatric bowenoid papulosis: risks and management. AB - A 34-month-old Haitian girl had a six-month history of asymptomatic vulvar papules. Histologic features and DNA probe studies were diagnostic of bowenoid papulosis. Resolution of the lesions was enhanced by cryotherapy. This is the third reported patient with prepubertal bowenoid papulosis. Sexual abuse should be suspected, but not assumed, in pediatric patients with this condition. Our patient had no evidence of sexual abuse. Bowenoid papulosis has been associated with cervical intraepithelial neoplasia and rarely with squamous cell carcinoma. Close gynecologic follow-up is recommended. PMID- 2559401 TI - Familial malignant retroperitoneal paraganglioma. AB - Paragangliomas are neuroendocrine tumours and those occurring in the head and neck have well recognized familial association. Retroperitoneal paragangliomas are uncommon and we present two cases of familial malignant retroperitoneal paraganglioma. Review of the literature revealed marked differences in the incidence and malignant potential of familial and non-familial paraganglioma. In contrast to the cases reported here, familial tumours are generally benign, though they may occur at multiple sites. Familial and non-familial paragangliomas may indeed be different disease entities. PMID- 2559400 TI - Mondor's disease associated with metastatic lung cancer in the breast. AB - We report what we believe is the first recorded case of Mondor's disease associated with metastatic lung cancer presenting in the breast. An association between Mondor's disease and primary breast cancer has been described previously. In patients with Mondor's disease and primary breast cancer has been described previously. In patients with Mondor's disease the possibility of underlying carcinoma, either primary or secondary, must be considered. PMID- 2559402 TI - Pulmonary vasculitis associated with cholangiocarcinoma of liver. AB - A 62 year old woman presented with an acute pulmonary vasculitis which responded to treatment with oral steroids. Investigations over one year revealed a cholangiocarcinoma of the liver. The association of vasculitis with neoplastic diseases remains a diagnostic challenge. PMID- 2559403 TI - The role of malignancy in lactic acidosis and shock. PMID- 2559405 TI - Malignant fibrous histiocytoma of sternum: an unusual cause of pyrexia of undetermined origin. AB - Malignant fibrous histiocytoma of the bone is a very rare tumour mainly affecting long bones, and the most common presenting symptom is local pain. A case of malignant fibrous histiocytoma presenting with pyrexia of undetermined origin and arising in the sternum, a localization not previously described, is reported. The patient died with septic shock after multi-drug chemotherapy including high dose methotrexate treatment with citrovorum factor rescue. PMID- 2559404 TI - Malignant fibrous histiocytoma. PMID- 2559406 TI - Malignant fibrous histiocytoma of the mediastinum. AB - A case of malignant fibrous histiocytoma of the mediastinum presenting with unusual features of fever and leucocytosis is reported. This is the youngest patient reported in the literature who had this tumour in the mediastinum. PMID- 2559407 TI - Reversible hypothyroidism detected by normal 99mTc scan. PMID- 2559408 TI - What is a histiocytoma? PMID- 2559409 TI - How safe is our blood? PMID- 2559410 TI - Sites of attachment to the nuclear scaffold in the human alpha and beta globin gene complexes. PMID- 2559411 TI - Topoisomerase II activity is not involved in the maintenance of active beta globin chromatin structure in avian reticulocytes. PMID- 2559412 TI - Identification and isolation of medicarpin and a substituted benzofuran as potent leukotriene inhibitors in an anti-inflammatory Chinese herb. AB - In a search for new inhibitors of leukotriene formation, a methylene chloride extract of the plant Dalbergia odorifera (Jiangxiang) was found to be a potent inhibitor of LTC4 formation in AB-CXBG Mct-1 mastocytoma cells. Following LH-20 and reverse phase HPLC chromatography, two compounds were isolated that had potent LTC4 inhibitory activity: medicarpin and 6-hydroxy-2-(2-hydroxy-4 methoxyphenyl) benzofuran (IV) with IC50s of 0.5 and 0.05 microM respectively. IV was shown to be a specific inhibitor of 5-lipoxygenase with an IC50 against the soluble rat enzyme of 0.08 microM, whereas it was inactive against cyclooxygenase. In neutrophils IV inhibited LTB4 production at comparable concentrations but had no effect on neutrophil degranulation or adhesion. PMID- 2559413 TI - Comparison of the effect of lipoxygenase metabolites of arachidonic acid and eicosapentaenoic acid on human natural killer cell cytotoxicity. AB - We compared in vitro effect of lipoxygenase (LO) products derived from arachidonic acid (AA) and eicosapentaenoic acid (EPA) on cytotoxic activity of human natural killer (NK) cell against human erythroleukemia cell line K-562. Leukotriene B4 (LTB4) derived from AA was found to significantly augment NK cell activity compared to the control level (in the absence of LTB). LTB5 showed a weak, but not significant, enhancing effect on NK cell activity. LTB4 was significantly more potent than LTB5 in the enhancement of NK cell activity. On the other hand, both 5- and 15-hydroperoxy fatty acids derived from AA and EPA significantly enhanced NK cell activity compared to the control level with similar potencies. PMID- 2559414 TI - The influence of antidepressive treatment on GABA-related mechanisms in the rat hippocampus: behavioral studies. AB - The effect of antidepressive treatment upon central GABAergic mechanisms has been studied in a behavioral model. Local injections (dentate gyrus of the dorsal hippocampus) of picrotoxin, a chloride channel blocker linked with the function of GABA-A receptor complex, potently stimulated rat motility recorded in the automated open fields. The intra-hippocampal administration of GABA antagonized the behavioral effect of picrotoxin (0.5 microgram). Similar effects were produced by addition to picrotoxin solution (0.25 microgram) of the GABA-A receptor agonist muscimol (0.25 microgram). Chronic (21-day), but not single, treatment of rats with desipramine (10 mg/kg, IP, daily) significantly attenuated picrotoxin-induced locomotor stimulation, when the GABA antagonist was given 24 hr after the last dose of the antidepressant. Repeated electroconvulsive shocks did not significantly change picrotoxin effect, while single shock produced some degree of inhibition of drug-induced motor stimulation. It is hypothesized that chronically applied desipramine, but not electroconvulsive shocks, may enhance the activity of hippocampal GABA-A receptor-related system. PMID- 2559415 TI - Genetic differences in opiate receptor concentration and sensitivity to ethanol's effects. AB - The hypothesis that genetic differences in opiate receptor concentration are involved in determining the sensitivity to some of the effects of alcohol was studied by comparing the hypothermic and analgesic effects of ethanol in four strains of mice that can be divided into three groups on the basis of their brain opiate receptor concentration: high (CXBH), low (CXBK) and intermediate (C57BL/6By and BALB/cBy). In the first experiment, animals within each strain were injected with either saline or 1.5 g/kg ethanol and their pain sensitivity was assessed 20 min later by the hot-plate test. The same procedure was repeated 10 days later with a higher dose of ethanol (2.5 g/kg). The lower dose produced analgesia only in CXBH mice, whereas the higher dose produced analgesia in CXBH, C57BL/6By and BALB/cBy mice, but had no effect in CXBK mice. In the second experiment, animals within each strain were injected with either saline or naloxone, followed 20 min later by an injection of 3.5 g/kg ethanol. CXBH mice were significantly more hypothermic and CXBK mice were significantly less hypothermic than all other strains. Naloxone attenuated ethanol's hypothermic effect in all strains except CXBK. These results suggest that the hypothermic and analgesic effects of ethanol are at least partly mediated by opiate receptors and are correlated with genetic differences in opiate receptor concentration. PMID- 2559417 TI - Effect of in vivo lithium treatment on (-)isoproterenol-stimulated cAMP accumulation in lymphocytes of healthy subjects and patients with affective psychoses. AB - Lithium treatment inhibited (-)isoproterenol-stimulated cAMP accumulation in lymphocytes of 83% of the normal control subjects investigated. Of the normal control subjects, 17% did not show inhibition of cAMP synthesis after lithium treatment. In one single normal control subject a clear inverse correlation was found between lithium serum concentration and maximal cAMP response. The cAMP levels of six manic-depressive and of four schizoaffective patients receiving lithium treatment did not differ significantly from those of 10 lithium-treated normal subjects. The results obtained in normal subjects seem to support the hypothesis that lithium reduces postsynaptic receptor sensitivity by decreasing cAMP response to agonist stimulation. Treatment outcome, however, does not seem to be exclusively related to a blunted noradrenergic response, since the only patient who did not respond to the treatment had a very low (-)isoproterenol induced cAMP response after lithium treatment. PMID- 2559416 TI - Chlordiazepoxide alters intravenous cocaine self-administration in rats. AB - This investigation was designed to examine the effects of benzodiazepines on intravenous cocaine self-administration in rats. Pretreatment with low doses of the benzodiazepine receptor agonist, chlordiazepoxide (0.3 to 1.0 mg/kg, IP), resulted in small but nonsignificant increases in drug intake with 0.5 mg/kg cocaine, while higher doses (10 mg/kg, IP) significantly decreased drug intake in all rats tested. The effects of chlordiazepoxide on self-administration were attenuated when the concentration of cocaine was increased to 1.0 mg/kg, suggesting that chlordiazepoxide was opposing rather than augmenting the pharmacological actions of cocaine. Pretreatment with the benzodiazepine receptor antagonist, Ro 15-1788 (1.0 to 10 mg/kg, IP), had no effect on self administration, suggesting that the reinforcing properties of cocaine do not result from direct interactions with benzodiazepine receptors. The result of this investigation demonstrate that chlordiazepoxide alters intravenous cocaine self administration in rats. Although additional research will be necessary to confirm these data, the results of this investigation suggest that chlordiazepoxide may decrease the reinforcing efficacy of cocaine through indirect actions on dopaminergic neuronal activity potentially mediated through GABAergic mechanisms via benzodiazepine receptor activation. PMID- 2559418 TI - The effects of pulsed, high frequency radio waves on rat liver (ultrastructural and biomedical observations). AB - The effects of a high frequency electro-magnetic field, generated by a Diapulse instrument (Diapulse Corporation of America) on rat liver has been investigated. Ultrastructural aspects are described and quantitative determinations of mitochondrial enzymes MAO, CyT-Ox, MDH, SDH and ATP-ase recorded. The standard therapeutic parameters generally used with the Diapulse instrument in medicine were found to induce a stimulation effect at the investigated level, without apparent degenerative modifications. A concordance between the qualitative ultrastructural data and quantitative subcellular enzymic determinations has been observed. PMID- 2559419 TI - Chronotropic effect of the methanolic extracts of the plants of the Paris species and steroidal glycosides isolated from P. vietnamensis on spontaneous beating of myocardial cells. AB - Rhizomes of five identified plants of the Paris species, Liliaceae, and Rhizoma Paridis which are sold as a crude drug named "Zao Xiu," "Qiyeyizhihua" or other names in nine different markets in China were tested for their effects on cultured cardiomyocytes. In the standard medium, eight methanol extracts out of sixteen at a concentration of 0.2 mg/ml stopped the spontaneous beating of myocardial cell sheets, but these extracts significantly increased the beating rate when the concentration was reduced to one half. In the culture medium with a low calcium concentration, 0.5 mM, the beating rate of the cells decreased to about 60% of that of the control in the standard medium. The addition of five of the extracts to the low calcium medium at a concentration of 0.1 mg/ml caused a stop of cell beating, but the other extracts increased beating rate at least by 10%. These steroidal glycosides isolated from the rhyzomes of P. vietnamensis (Takht.) H. Li also stimulated cell beating. Among them, diosgenin-3)-alpha-L rhamno-pyranosyl-(1---2)-(alpha-L-arabinofura nosyl-(1---4))-D-glucopranoside (compound 1) was the most effective stimulant for cell beating as well as calcium uptake by the myocardial cells. PMID- 2559420 TI - The antiviral action of lignans. AB - A total of 18 purified lignans was evaluated for antiviral activity against murine cytomegalovirus (CMV) and Sindbis virus, by means of different treatment regimens. Podophyllotoxin and alpha-peltatin were the most potent compounds, and they apparently inhibited murine CMV at an essential early step in the replication cycle after the adsorption of virus to the cells. On the other hand, justicidin B and the diphyllin derivatives were much more effective against Sindbis virus, and 12 of the lignans had no demonstrable effect at all, despite their known activities in other bioassays. PMID- 2559421 TI - 3H-imipramine binding in the frontal cortex of suicides. AB - Imipramine binding (desipramine- and serotonin-sensitive) was determined in the frontal cortex of suicide victims and nonpsychiatric controls who died due to medical disease or accidents. There were no differences in Kd or Bmax of imipramine binding between controls and suicides. The Kd and Bmax values of serotonin-sensitive imipramine binding were significantly lower than desipramine sensitive imipramine binding, both in controls and suicides. There were significant correlations between Kd and Bmax of serotonin-sensitive imipramine binding and desipramine-sensitive imipramine binding in suicides but not in controls. PMID- 2559422 TI - Platelet adrenoceptors and prostaglandin responses in depressed patients. AB - Platelet alpha 2-adrenergic receptor binding and prostaglandin responsivity were measured in depressed patients. Depressed patients had significantly higher platelet 3H-dihydroergocryptine (3H-DHE) binding values than controls. Depressed patients also showed significantly reduced prostaglandin E1-stimulated cyclic adenosine 3',5'-monophosphate (cAMP) production and significantly decreased % inhibition of cAMP production by norepinephrine. These results support the suggestion that there may be a dissociation between alpha 2-adrenergic receptor binding and responsivity in depression. There were no significant correlations between platelet adrenergic variables and other indices of noradrenergic function. However, there was a significant correlation between 3H-DHE binding values and basal plasma levels of cortisol. PMID- 2559423 TI - Protection by WR-3689 against gamma-ray-induced intestinal damage: comparative effect on clonogenic cell survival, mouse survival, and DNA damage. AB - The aminophosphorothioate WR-3689 was characterized for its ability to protect mouse jejunal cells in vivo from single doses of X or gamma radiation. First, the effect of the drug on the survival of jejunal stem cells was examined using a clonogenic end point, the crypt microcolony assay. When WR-3689 was administered 30 min prior to whole-body irradiation, the number of surviving crypt cells was markedly increased at all doses of the drug, although protection began to level out at doses larger than 600 mg/kg. Protection was maximal when the drug was given 30 min before whole-body irradiation and declined rapidly with both shorter and longer intervals. Protection factors (PFs) were obtained by measuring survival curves for clonogenic crypt cells as a function of radiation dose; WR 3689 given 30 min before whole-body irradiation protected jejunum in the microcolony assay with a PF of 1.26 +/- 0.02, 1.50 +/- 0.10, and 1.65 +/- 0.10 at doses of 200, 400, and 800 mg/kg, respectively. Next, the effect of WR-3689 on the survival of jejunal stem cells was determined by assaying the survival of mice given X-ray doses to the whole abdomen in the range leading to death from the gastrointestinal syndrome. The PFs based on the LD50 values for 11-day survival were 1.31 +/- 0.05 (200 mg/kg) and 1.48 +/- 0.05 (400 mg/kg). Crypt-cell survival and animal survival were thus modified to a similar extent by this agent. Finally, the effect of WR-3689 on the induction of DNA single-strand breaks (SSBs) in jejunal cells was measured using an adaptation of the alkaline elution methodology. In mice treated with WR-3689 (400 or 800 mg/kg) 30 min prior to whole-body irradiation with 10 Gy there was no significant reduction in the number of DNA SSBs induced either in samples of the jejunum or in the cycling crypt cells, providing further evidence that there is no simple relationship between the modification of DNA SSBs and the survival of jejunal stem cells. PMID- 2559424 TI - [The effect of external gamma-irradiation on the distribution of deuterium oxide in the rat body]. AB - External gamma-irradiation (7.74 and 15.48 x 10(-2) C/kg) does not influence the distribution of deuterium oxide within a rat body: it is distributed uniformly among the organs and tissues as it is observed in nonirradiated animals. The effect of external irradiation favors the retention of deuterium oxide within the organs and tissues. The delayed excretion of deuterium oxide from the body can enhance the biological effect. This should be taken into account in standardizing the combined radiation effects. PMID- 2559425 TI - [Use of sarcoma 180 TG cells for obtaining ascitic fluid from mice hyperimmune to the foot-and-mouth disease virus]. AB - Mice immunized with FMDV C3 Arg 84 antigen were inoculated intraperitoneally with sarcoma 180 TG. The ascitic fluid obtained by ventral puncture contained high titers of antibodies, similar to those obtained from serum, as determined by neutralization and ELISA tests. Ascitic volumes were 10 to 20 times greater than those obtainable with. PMID- 2559426 TI - [Rapid serologic diagnosis of Argentinian hemorrhagic fever in whole blood]. AB - The usefulness of a method for detection of antibodies against Junin virus in whole blood was tested. N: NIH adult mice were inoculated with 10(3) PFU of attenuated XJ-Clon 3 Junin virus strain by intraperitoneal route and blood was obtained by retro-orbital puncture at 21 days post-infection. One blood aliquot (50 microliters) was collected in tubes containing a stabilizer solution for whole blood and another was processed for serum obtention. Immunofluorescent antibodies were tested on spot slides of a BHK/21 cell line persistently infected with Junin virus. High antibody titers (1/64 to 1/256) were detected in both whole blood and serum, with 66% coincidence between both procedures. These results show that the method of detection of antibodies in whole blood would be useful to test quickly for anti-Junin virus antibodies in seroepidemiologic studies, in endemic areas. PMID- 2559427 TI - [Regulation of intracellular pH by Na+/H+ exchanger]. PMID- 2559428 TI - [Mechanism for energy dependence in intracellular protein breakdown]. PMID- 2559429 TI - Small cell lung cancer: single agents revisited. PMID- 2559430 TI - [Our experience in testing the lacrimal drainage system using radionuclides (nuclear dacryocystography with 99mTc O4)]. AB - The function of the lacrimal drainage system was tested in 5 patients and 3 controls by scintigraphy with a Pho-GAMMA HP-1206 Nuclear scanner Chicago, coupled with a visualization system and data processing device of type MB 9101 GAMMA. PMID- 2559431 TI - [Data on ophthalmologic diseases during pregnancy]. AB - The authors present the palpebral, uveal, neurologic, retinal manifestations and mainly diabetic retinopathy during pregnancy. Likewise, the paper analyses the relationship: high myopia, detachment of the retina und pregnancy, and the therapeutic attitude at ophthalmologic and obstetrical levels. PMID- 2559432 TI - [Oat cell carcinoma of the esophagus]. AB - Extrapulmonary locations of undifferentiated small cell carcinoma of the "oat cell" type are uncommon. In spite of its rarity, the appearance of this type of tumor has been described in various organs of the economy. Of these, the esophagus is the most common location, constituting 0.05 to 7.6% of esophageal carcinomas. Of uncertain histopathologic origin, these tumors are characterized by their extreme aggressiveness, which carries a poor short term prognosis. Surgery, radiotherapy and chemotherapy, separately or in combination, have produced discouraging results in the treatment of primitive esophageal "oat cell" carcinoma. In this paper are presented four cases of esophageal "oat cell" carcinoma treated in the Hospital de la Santa Cruz y San Pablo in 1979-1988, out of a total of 260 carcinomas in the same location. The clinical evolution and pathological findings of this type of tumor are commented. The literature on this unusual histological type of carcinoma is reviewed. PMID- 2559433 TI - [Glomus tumor of the stomach. Optic and immunohistochemical study. Apropos of a case]. AB - A case of glomus tumor of the stomach occurring in a patient of 59 year old founded during a cholecystectomy is described. Clinicopathological and immunohistochemical findings are commented. PMID- 2559434 TI - Lack of effect of alpha-tocopherol and ascorbic acid on the response of some physiological variables to immobilization stress in rat. AB - The effect of alpha-tocopherol or ascorbic acid jointly and separately on the seric levels of adrenocorticotropin (ACTH) and corticosterone has been studied. ACTH response to stress is similar in all groups, whereas the corticosterone response is higher in the animals treated with alpha-tocopherol. Neither lactate response nor gastric ulceration caused by stress was influenced by the administration of alpha-tocopherol and/or ascorbic acid. These results suggest that free radicals might not be implicated in the control of the physiological response to stress. PMID- 2559435 TI - Structural and functional characterization of the OmpF and OmpC porins of the Escherichia coli outer membrane: studies involving chimeric proteins. AB - The roles of submolecular regions of OmpF and OmpC, major outer membrane proteins of Escherichia coli, as concerns their biogenesis, structure and function were studied using a large number of chimeric genes constructed from the ompF and ompC genes through single or double homologous in vivo recombination. When recombination between the two genes took place at certain regions of their central regions, no chimeric protein was detected, irrespective of whether the amino-terminal and carboxy-terminal regions were derived from OmpF or OmpC. Biochemical studies revealed that these proteins were synthesized and exported across the cytoplasmic membrane normally, but that they were not properly assembled into the outer membrane and hence were degraded rapidly. Characterization of these chimeric proteins, in which recombination between OmpF and OmpC took place once or twice, suggested that the central region of each of these proteins plays an important role in the respective assembly, whereas the roles of the amino-terminal and carboxy-terminal regions may be marginal. Functional characterization of these chimeric proteins revealed the regions important for the receptor functions of OmpF and OmpC for phages TuIa and TuIb, respectively. PMID- 2559436 TI - Nucleotide sequence of the plasmid-borne virulence gene mkfA encoding a 28 kDa polypeptide from Salmonella typhimurium. PMID- 2559438 TI - Bone marrow peroxidase in mice: determination of the level and the catalytic activity. AB - The level of myeloperoxidase and the peroxidase activity in the bone marrow were determined in the various strains of mice. The levels of myeloperoxidase, which was determined by a immunostaining coupled with sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE), were ranged from 3 to 5 micrograms per mg protein in the bone marrow of various strains of mice. No strain difference of the enzyme activity toward guaiacol was observed among mice. On the other hand, the activity toward KI of C3H/He mice was lower than C57BL/6, Balb/c and ddY mice. PMID- 2559439 TI - Differential interaction of guanabenz with receptor binding sites in rat brain and kidney. AB - The alpha 2-adrenoceptor selective agonist, [3H]guanabenz ([ 3H]GBZ), labels a unique population of binding sites in whole kidney which are not labeled by [3H]p aminoclonidine ([3H]PAC). These binding sites are saturable and of high affinity (Kd = 10-12 nM). [3H]GBZ was not displaced from these sites by other alpha 1- or alpha 2-ligands, suggesting that they are non-adrenergic. This hypothesis is further supported by the insensitivity of renal guanabenz binding to regulation by guanyl nucleotides or to destruction by trypsin. Also, there appears to be no effect of guanabenz on the potency of isoproterenol in competing for beta adrenoceptors in the kidney, which has been previously reported to be sensitive to clonidine. The absence of any effect of guanabenz on isoproterenol displacement of [3H]dihydroalprenolol in kidney suggests there are subtle differences in activation of alpha-receptors by clonidine and guanabenz in the kidney. In the brain, [3H]GBZ labels two binding sites. Part of the binding of [3H]GBZ in the brain is to sites essentially identical to the alpha 2 adrenoceptors labeled by [3H]PAC. The remainder of the binding resembles the non adrenergic binding in kidney. The relationship of this unique binding site to the pharmacologic actions of guanabenz is currently not known. PMID- 2559437 TI - Erythrocyte receptors for cholera and heat-labile enterotoxins of Escherichia coli. AB - Many serological reactions using red blood cells (RBC) such as radial immune haemolysis (RIH) and indirect haemagglutination (IH) tests have often been used for the detection of cholera toxin (CT) and heat-labile (LT) enterotoxin produced by porcine and human Escherichia coli strains. In these tests, the enterotoxins bind to sheep, bovine and guinea-pig RBC without any ligand. We studied several factors which might interfere with such binding, as well as the nature of the receptors involved. Treatment of erythrocytes with different enzymes revealed that proteolytic enzymes had no effect on the adsorption of enterotoxins to RBC. Conversely, treatment with neuraminidase increased the adsorption. Experiments carried out with delipidized RBC revealed that none of the enterotoxins under study bound to the cells thus treated. Pre-incubation of ganglioside fractions with the enterotoxins blocked RIH and IH reactions and the biological effect of them on Vero cells. Assaying RBC ganglioside fractions by thin-layer chromatography revealed the presence of GM1. Our results suggest that the receptors for GT and LT enterotoxins in sheep, bovine and guinea pig RBC are gangliosides: mainly GM1. PMID- 2559442 TI - Characterization of a 75-kDa Epstein-Barr virus capsid protein using a new monoclonal antibody H250. AB - A monoclonal antibody (mAb) designated H250, directed against an Epstein-Barr virus (EBV) capsid antigen, was obtained following immunization of BALB/c mice with naked particles from the producer cell line B95.8. This antigen was present in the producer lines B95.8, P3HR1, M81, RI and CA, and absent from the non producer lines BJAB, Raji and 1022. H250 did not inhibit the transformation of cord blood lymphocytes by the B95.8 virus, nor did it inhibit EA induction on Raji cells by the P3HR1 virus. In addition, H250 showed no fluorescence on living B95.8 cells. This indicates that H250 does not recognize a membrane antigen. By indirect immunofluorescence, no fluorescence was observed on induced Raji cells or on PAA-treated B95.8 cells. Thus, H250 recognized a late antigen of the EBV virus replication cycle. Agglutination of naked virus by H250 showed it was directed against a capsid antigen. Positive fluorescence was observed on cells treated with tunicamycin, indicating that H250 recognized a protein. The molecular weight of this protein was obtained by Western blot and was approximately 75 kDa. The blocking tests carried out with H250 seemed to indicate that this Ab appeared late in patient sera during primary infection. PMID- 2559440 TI - Oxidation of diethyldithiocarbamate to disulfiram by liver microsomal cytochrome P-450-containing monooxygenase system. AB - We examined the involvement of cytochrome P-450 in the oxidation of diethyldithiocarbamate (DTC) to disulfiram (DS) by liver microsomes in the presence of NADPH. DS difference spectra of liver microsomes showed a peak and trough at about 385 and 418 nm, respectively, which disappeared after further addition of glutathione (GSH). DTC alone had little effect on the microsomal spectrum, however, the addition of NADPH gradually produced a spectral change having a trough at 416-417 nm, which waned upon further addition of GSH. Microsomal DS production was increased by phenobarbital pretreatment and decreased by carbon tetrachloride pretreatment, depending on the activity of the cytochrome P-450-monooxygenase system. With microsomes peroxidized by cumene hydroperoxide, the extent of NADPH-dependent DS production lowered in proportion to the decrease in cytochrome P-450. Inhibitors of cytochrome P-450 such as SKF 525A, metyrapone and n-octylamine dose-dependently inhibited the DS production. Sodium azide, an inhibitor of catalase, increased the DS production, whereas addition of exogenous catalase only slightly suppressed it. It is concluded that oxidation of DTC to DS by liver microsomes largely proceeds via the cytochrome P 450-containing monooxygenase system and partly by hydrogen peroxide generated during NADPH oxidation. PMID- 2559441 TI - Genetic stability of poliovirus insertion mutants with a foreign oligopeptide on the capsid surface. AB - The genetic stability of poliovirus mutants which carry a foreign oligopeptide on the surface of their capsid was studied (1) upon mutant isolation, (2) after serially diluted passages in cell cultures, and (3) in persistently infected cultures which have been recently developed. Viruses having a 3-codon insertion within the VP1 capsid protein-encoding region appeared to be extremely stable, except in the specific case of persistent infection. Viruses having a 6-codon insertion were slightly less stable. Point mutations and one recombination event were observed as soon as viruses were recovered and studied following plasmid transfection. Additional point mutations appeared within the insertion after 12 serially diluted passages in monkey kidney cells. Under all test conditions, the foreign insertion was never deleted from the virus genome. PMID- 2559443 TI - [A periventricular tumor associated with acute kidney failure and bilateral renal cortical necrosis]. AB - The authors present a case in which the clinico-evolutive particularity was an association between a periventricular tumour (multiform glioblastoma) and acute renal failure. Considerations are made on clinical signs determined by the neoplastic process (apraxia-agnosia hyperthermia, etc.), and on the disseminated intravascular coagulation syndrome related to the acute renal failure. PMID- 2559444 TI - Use of low-angle diamond knives leads to improved ultrastructural preservation of ultrathin sections. AB - Low-angle diamond knives, one of them with an angle as low as 14.6 degrees, have been tested to measure compression induced during the sectioning of polystyrene latex spheres embedded in epoxy resins. Compression diminishes roughly proportional to the sectioning angle (sum of knife and clearance angles). The beneficial application of low-angle diamond knives in biology and material science is illustrated by sections through a muscle fiber (compression reduced by a factor of 3), sections through a collagen fiber (meridional resolution improved by a factor of 2) and sections through a copper grid bar (reduced curling). These low-angle diamond knives are as resistant as the classical knives having angles of 45 degrees and are routinely used by the author. Low-angle diamond knives with angles down to 35 degrees are now commercially available. PMID- 2559445 TI - Probe size and bound label conformation in colloidal gold-ligand labels and gold immunolabels. AB - Colloidal gold can be produced in sizes ranging from 1.0nm to 150nm. All sizes of gold can be conjugated, principally by hydrophobic bonding, to a variety of molecules including ligands, enzymes and antibodies, as well as lectins and polysaccharides. The activity of most of these biological molecules is retained on conjugation with gold particles irregardless of size range, although the ratio of protein surface area to gold particle surface area varies widely depending on particle and protein size. We have employed low voltage high resolution scanning electron microscopy to compare, microscopically, the shapes of biological molecules unbound, bound to very small (3nm) gold particles, and bound to larger (18nm-30nm) gold particles. When very small gold particles are conjugated to large protein molecules, several particles bind along the length of each molecule, while smaller protein molecules often wrap around a single small gold particle. With larger gold particles, several biological molecules bind to a single gold particle. In addition, the shape of protein molecules bound to larger gold particles differs from that of molecules bound to small gold particles. PMID- 2559446 TI - Deoxyribonucleic acid sequence mapping on metaphase chromosomes by immunoelectron microscopy. AB - Nucleic acid sequences can be localized on chromosomes in the electron microscope after hybridization with a biotinylated DNA probe followed by detection with a primary antibiotin antibody and a secondary antibody coupled to colloidal gold. Hybridization probes can also be labelled with alternative ligands such as N acetoxy-2-acetylaminofluorene (AAF), Dinitrophenyl-dUTP and Digoxigenin-dUTP. Multiple labelling is possible if these differently modified DNA probes are used in conjunction with colloidal gold preparations of varying particle sizes. A substantial signal amplification can be achieved by incubating preparations with successive cycles of primary antibiotin antibody followed by a biotinylated secondary antibody. Detection is with Streptavidin-gold, and in the case of highly and moderately repeated sequences, the signal is visible in the light microscope. Detailed protocols are given for EM in-situ hybridization to whole mount metaphase chromosomes and include instructions necessary to perform multiple sequence localization and signal amplification. PMID- 2559447 TI - The organization and substructure of chromatin fibres in the interphase nucleus as studied by scanning electron microscopy. AB - The high packaging ratio of DNA in both interphase nuclei and metaphase chromosomes presents great difficulties to our understanding of the three dimensional organisation of processes such as DNA replication and transcription in the nucleus. Although the higher order structure of DNA, in terms of the way it is organised into the unit fibre of chromatin has received much attention over the last decade, the highest levels of packaging of chromatin in both nuclei and chromosomes have hardly begun to be elucidated. Much of the difficulty in investigating fibre organisation with conventional methods is the inherent two dimensional nature of sectioned or spread material in the transmission microscope. Three dimensional imaging from the SEM has, until recently, been limited by the available resolution. Our own previous studies of chromosome structure have shown that a combination of 'in lens' imaging combined with the high signal generation imparted by osmium impregnation have been adequate to routinely visualise chromatin fibre organisation in metaphase chromosomes, and the changes that occur as a result of a variety of banding techniques. More recent experiments using the same techniques on interphase nuclei extracted from a variety of tissue culture cells have indicated that Scanning electron microscopy of nuclei is a potentially useful technique for studying chromatin organisation, which may be made more accessible by a variety of biochemical extraction methods. PMID- 2559448 TI - [Kearns-Sayre syndrome: mitochondrial encephalomyopathy caused by deficiency of the respiratory chain]. AB - We report the cases of a 46 year old woman and of a 18 year-old boy who met the criteria for Kearns-Sayre syndrome. Additional atypic features were present in one case: family history, psychosis and acute respiratory failure. In both cases histoenzymatic analysis of the muscle biopsy and biochemical studies of mitochondria isolated from the muscle sample demonstrated mitochondrial myopathy associated with combined partial deficiency of complexes I and IV of the electron transfer chain. Although there is no correlation between clinical and biological data in the mitochondrial myopathies our cases confirm that such defects are involved in Kearns-Sayre syndrome. Improvement with coenzyme Q10 therapy in these patients is reported. PMID- 2559449 TI - [Painful legs and moving toes. Neuropathy caused by cytarabine]. AB - A new case of painful legs and moving toes syndrome is reported in a patient with a neuropathy due to high-doses of cytosine arabinoside (Ara C). A few cases of this complication have been reported. The neuropathy was sensory, axonal, and incompletely regressive. Carbamazepine had a dramatic effect on pain and involuntary movements. PMID- 2559450 TI - [Amyloid pseudotumor of the sciatic nerve]. AB - A 60 year-old man complained of numbness and pain in the right lower limb, suggesting lesions of the fifth lumbar and first sacral roots. Sixteen months later, CT showed a tumor of 3.5 cm at the emergence of the first right sacral root. Microscopic examination disclosed an infiltration of the fibers of the nerve by numerous masses of hyaline eosinophilic material which stained with Congo red and produced green birefringence under polarized light. The persistence of congophilic properties of the amyloid deposits after permanganate pretreatment suggested an immunoglobulin origin (AL). A research of amyloid deposit in others viscera: heart, kidneys, digestive tract, was negative. We believe that this is the first reported case of amyloid pseudo-tumor involving a peripheral nerve. PMID- 2559451 TI - [Cushing syndrome due to ectopic ACTH secretion]. AB - The authors studied 8 patients (4 males and 4 females) with Cushing's syndrome due to ectopic ACTH secretion. Chronological age ranged from 15 to 45 years and duration of the disease ranged from 3 to 48 months. All patients presented typical signs of Cushing's syndrome, blood hypertension, and four of them had hyperpigmentation of the skin. Five patients had fasting hyperglycemia and all patients but one had serum hypokalemia (serum K = 2.2 to 3.9mEq/l). The circadian rhythm of cortisol was absent in all patients and basal cortisol levels were elevated in all patients but one. Basal ACTH levels evaluated in 7 patients were elevated in 6 (29 to 1050 pg/ml-MRC). One patient presented normal depression of urinary 17-OH after two days of dexamethasone and normal increase of urinary 17 OH and serum 11-dexycortisol after methyrapone. Four patients had carcinoid tumor (3 thymic and 1 bronchial), two had pancreatic islets cell tumors, one had bilateral pheochromocytoma and medular carcinoma of the thyroid, and one had oat cell carcinoma of the lung and medular carcinoma of the thyroid. Thoracic X-rays identified the ectopic ACTH secretion tumor in four cases, all confirmed by CT scan. Abdominal CT showed a difuse enlargement of the adrenals in seven cases and bilateral nodules in one case (pheochromocytomas). Six patients died within 3 years of the diagnosis. The authors concluded that clinical and hormonal findings could mislead the findings of ACTH ectopic secretion and Cushing's disease, and suggest that thoracic X-rays and CT scans of the skull, thorax, and abdome should be done in all cases of Cushing's syndrome. PMID- 2559452 TI - [Multiple familial polyposis. Report of 4 cases and association with mucinous adenocarcinoma of the colon]. AB - Four cases of familial polyposis coli (FPC), seen at Dr. Manuel Gea Gonzalez General Hospital from 1985 to 1987, are presented. Three females and one male, with ages ranging between 23 and 52 years, are presented. The clinical picture included bloody feces, polyp prolapse, and a maxillary osteoma. In 3 cases the correct preoperative diagnosis was made by rectosigmoidoscopy, biopsy and barium enema. In one patient, urgently operated on, the diagnosis was postoperatively made by colonoscopy. The adenocarcinoma of the colon appeared associated with FPC in three cases (one of the mucinous variety). One total coloproctectomy and two abdominal colectomies with ileorectal anastomosis were performed. One patient operated on because of bowel obstruction was treated by right hemicolectomy. This case developed abdominal carcinomatosis afterwards. One woman died; another one was lost during follow-up, and the other one had no complications. The association of FPC and mucinous adenocarcinoma of the colon is reported for the first time. PMID- 2559453 TI - [The current status of the pathogenesis of sarcoidosis]. PMID- 2559454 TI - [The spread of sarcoidosis in the Socialist Republic of Romania]. PMID- 2559455 TI - [Points of view on the treatment of sarcoidosis]. PMID- 2559457 TI - [The nosologic picture of sarcoidosis]. PMID- 2559456 TI - [The process of the formation of sarcoid granuloma]. PMID- 2559458 TI - [Current clinical aspects of sarcoidosis]. PMID- 2559459 TI - [Assessment and prognostic criteria in mediastino-pulmonary sarcoidosis]. PMID- 2559461 TI - [Morphologic diagnosis of sarcoidosis by transbronchial puncture]. PMID- 2559460 TI - [Clinico-radiologic observations in mediastino-pulmonary sarcoidosis]. PMID- 2559462 TI - [Serum enzyme tests in the diagnosis and follow-up of the evolution of sarcoidosis]. AB - Due to the diagnosis difficulties posed by sarcoidosis, new metabolic or biological tests were suggested for replacing and/or confirming the histopathological examination: determination of the serous levels of the angiotensin-convertase (ACS), lysozyme and acid phosphatase. For establishing the ACS diagnosis value, the enzyme serous level was determined in 117 sarcoidosis patients, histopathologically confirmed (62 with active sarcoidosis, 55 with chronic sarcoidosis) in comparison with 109 patients suffering from other chronic lung affections. The results obtained, statistically processed, showed that the ACS level in active sarcoidosis is increased in 97% of the patients and only in 8% of the patients non-suffering from sarcoidosis. In active sarcoidosis, the ACS level has a positive predicting value of 86.8% and a negative one of 98%. The serous activities of lysozyme and acid phosphatase are not significant in sarcoidosis diagnosis. Of 59 treated patients with active sarcoidosis, in 58 the ACS value became normal after the clinical-radiological remission, thus showing the prognosis value of the test. PMID- 2559463 TI - [The advantages of Reprimum therapy in pulmonary sarcoidosis and other granulomatous diseases]. AB - In sarcoidosis and other granulomatous non-caseous diseases, the election treatment is immunosuppressive, mainly with cortisones that ensure more than 70% lasting remissions. Continuous use of cortisones for a long time (8-30 months) in high doses leads to serious side effects: gastric and intestinal ulcers, obesity, osteoporosis, suprarenal dysfunction, sensitivity to infections. Good results and elimination of the important side effects were obtained by treatment with Reprimum--a semisynthetic antibiotic with a wide spectrum and immunosuppressive properties--administered alone or with prednisone in small doses (15-20 mg once) in 6 weeks' series: 2 weeks--Reprimum 10/mg/kg daily +/- prednisone and for other 4 weeks--Reprimum 15 mg/kg twice a week +/- prednisone followed by two weeks' break. In 75 patients with histopathologically confirmed sarcoidosis (of whom 7 9.3% with outside-the-lung situs, too), the treatment with Reprimum gave: 94.7% lasting remission, only 5.3% failures, reduction of the treatment period to 6-12 months and the absence of any important side reaction. In other 37 sarcoidosis cases, failures of cortisone therapy (of which 11-30% relapses after 2-6 years), the treatment with Reprimum together with prednisone allowed recovery of 29 patients (78.4%). The same treatment with Reprimum, used in 22 patients with immunosuppressive treatment indication (dermatomyositis, Kaposi's syndrome, thrombocytopenias, nodose periarteritis, silicosis), of whom 18 (81.8%) were failures of the cortisone therapy, healed 20 of these cases (90.9%). Reprimum immunosuppressive property acts at the level of T4+ lymphocyte, involved in sarcoidosis pathogenesis. The functional blockage of T4+ lymphocyte can be also achieved by cyclosporine A.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559464 TI - [A comparative study of plasma converting enzyme and renin in sarcoidosis]. AB - The paper reports on a comparative study of the conversion enzyme and plasmatic renin performed on a group of 55 patients, with clinical diagnosis of sarcoidosis and other lung diseases. Whereas the conversion enzyme showed important increases in active sarcoidosis and a tendency to the normal state after treatment with corticoids, the activity of the plasmatic renin remained the same, both before and after corticotherapy. The result obtained prove the practical usefulness of the dosage test of the conversion enzyme, in agreement with the literature data. They also point to the possible participation of the pulmonary SRA in the vascular and local enzymatic-chemical imbalances, suggesting the possibility of associating the conversion enzyme inhibitors to the treatment of sarcoidosis with corticoids. PMID- 2559465 TI - [Segmental bronchoalveolar lavage with a flexible probe via a rigid bronchoscope in the diagnosis of mediastino-pulmonary sarcoidosis]. AB - Segmentary bronchial-alveolar lavage with flexible catheter connected to rigid bronchoscope might be used in diagnosis of mediastinal-pulmonary sarcoidosis, according to a method used by the authors. Cytologic examination of the lavage fluid shows, in the cases studied, the great abundance in cells, with lymphocytes increase to 30 +/- 12% (in agreement with other authors) making thus possible the disease diagnosis. On the other hand, polymorphonuclears increased to 20 +/- 10%. These higher values than those noticed up to now, pointed that the fibrosing process in sarcoidosis has a higher level than assumed, and appear since the onset of the disease. PMID- 2559467 TI - [The ENT examination in the diagnosis of sarcoidosis]. PMID- 2559466 TI - [Diagnostic errors in sarcoidosis]. PMID- 2559468 TI - [The efficacy of transbronchial puncture in sarcoidosis]. PMID- 2559469 TI - [Acute hemorrhagic conjunctivitis caused by a variant of coxsackievirus A24, in Belem, Para, Brazil, 1987]. AB - An epidemic of acute haemorrhagic conjunctivitis in Belem, Para, Brazil, was investigated. From 83 patients, 73 samples of virus was isolated in cultures of HEP-2 cells from conjunctival swab and throat swab. The virus isolates were identified in Centers for Disease Control, Atlanta, Georgia, USA, as an antigenic variant of coxsackievirus A24. Neutralization Test was done on 56 paired serum samples, using entero virus type 70 (EV70) and virus isolates. Serological conversions for virus isolates were found in 57% (32 patients). PMID- 2559470 TI - [Black grain mycetoma caused by Madurella grisea]. AB - The authors reported one case of eumycotic mycetoma due to Madurella grisea (black grains) occurred on the right foot of the patient studied. The structure, microscopic morphology and therapeutic evolution are also studied and reported. PMID- 2559471 TI - [Late motor response: axon motor reflex and peripheral late wave]. AB - The axon reflex (AR) is an intermediate response evoked by submaximal stimulation and eliminated by supramaximal stimulation: peripheral late wave (PWL) is a late motor response that appears with supramaximal stimulation and is regularly repeated. The PWL has been studied in 185 subjects and has been obtained in 70 cases (36 cases of polyneuropathies, 23 of nerve root lesions, 6 of peripheral nerve lesions, 1 of poliomyelitis and 4 healthy subjects); the AR has been studied in 35 subjects and found in 4 cases (2 cases of nerve root lesions, 1 of poliomyelitis and 1 of peripheral lesion). It seems that the two responses have no common characteristics and that the persistence of the PWL is rather inconstant. PMID- 2559472 TI - Ocular onchocerciasis and intensity of infection in the community. II. West African rainforest foci of the vector Simulium yahense. AB - A novel method of analysis was used to describe community patterns of ocular onchocerciasis in relation to the intensity of infection in West African forest villages where S. yahense is the sole vector. The pattern is completely different from that found in the savanna, even after correction for the intensity of infection as measured by the Community Microfilarial Load (CMFL). Lesions of the anterior segment of the eye as well as onchocercal blindness either do not occur or occur only sporadically with increasing CMFL in the Yahense forest whilst a steep linear relation exists between the prevalence of these lesions and the CMFL in the savanna. Lesions of the posterior segment of the eye are also less common in the Yahense forest. For a given skin microfilarial load, the ocular microfilarial load is lower in the Yahense forest. For a given ocular microfilarial load, a lower prevalence of eye lesions is found in the Yahense forest compared to the savanna. It is concluded that microfilariae of Onchocerca volvulus in the Yahense forest are less eye invasive than microfilariae from the savanna. Furthermore, they appear to be also less pathogenic to the eye. These findings explain why ocular onchocerciasis is relatively mild in the Yahense forest, in spite of the high intensities of O. volvulus infection in the community. PMID- 2559474 TI - [Blood levels of ACTH and endogenous digitalis-like substances in in man]. AB - Simultaneous determination of immunoreactive ACTH and immunoreactive digitalis like substances (DLS) in 71 plasma specimens of 44 persons proved that in spite of a great range of ACTH concentrations in the followed individuals (1.7-271 nmol/l) there is not a major correlation between DLS and ACTH or a significant difference in plasma DLS concentrations in the groups of persons with suppressed, normal or increased ACTH concentrations. Acute increase of ACTH plasma concentrations after synthetic ACTH application or a suppression of plasma ACTH by dexamethasone were not accompanied by corresponding changes of immunoreactive DLS. Thus it follows from this investigation that in spite of its biological digitalis like activity and interference of synthetic ACTH 1-24 high concentrations with digoxin enzymoimmunoassay in vitro, ACTH in concentrations found in human plasma is not responsible for endogenous immunoreactive DLS levels in plasma. PMID- 2559473 TI - [The effect of exposure to silicon oxides on the occurrence of porphyria cutanea tarda]. AB - The incidence of porphyria cutanea tarda was studied in two groups of silicon dioxide risk workers and compared to that in a control group. In the group with higher exposure to SiO2 the illness occurred in 12 out of 440 workers, in the group with lower exposure in 12 out of 1000 workers. In the control group porphyria was found in 18 out of 12,100 individuals examined. The difference in the incidence of the illness between the SiO2 risk individuals and the control group is statistically significant (p less than 0.01), while the difference between the group with variously high SiO2 exposure is on the level of statistical significance (p less than 0.05). The causes of the higher incidence rate of porphyria cutanea tarda in free SiO2 risk workers are discussed. PMID- 2559475 TI - Occurrence of specific receptors in the duodenum. AB - Three groups of receptors defined pharmacologically by means of specific agonists and competitive antagonists were described: receptors for the so-called gastrointestinal hormones, receptors for the autacoids and receptors for the neuromediators. Together with the better known post-synaptic receptors pre synaptic ones were also discussed in the light of the different possible interactions between one and the other. The mode of action of drugs stimulating (agonists) or inhibiting (antagonists) the same receptors is so far a very intriguing problem. The availability of newer and more selective compounds capable of distinguishing not only different kinds of receptors but also the subtypes of a single class of receptors will certainly contribute to better understanding of this situation. PMID- 2559476 TI - Evaluation by immune scanning electron microscopy of foscarnet treatment of cytomegalovirus infection in patients with renal transplants. AB - Eight patients, 7 renal and 1 combined renal and pancreas allograft recipients with generalized cytomegalovirus (CMV) infection were treated with continuous intravenous foscarnet infusion (0.15 mg/kg/min) for 10-14 days. Antiviral effect was studied by immune scanning electron microscopy for detection of CMV antigen in serum and urine, by virus isolation in tissue culture in samples from buffy coat, broncholavage and urine and by serology. CMV antigen was detected in serum samples in 8 patients and 5 had positive virus isolation from buffy coat, before institution of therapy. The 3 patients with negative virus isolation in tissue culture had serological evidence of a reactivated CMV infection. Virus replication was inhibited by foscarnet treatment in 7 patients within a week (p less than 0.01) (Wilcoxon log rank test). 7/8 patients had no detectable CMV antigen in serum or urine after 7-10 days of treatment (p less than 0.01) (Wilcoxon log rank test). PMID- 2559478 TI - Retroviruses and chronic arthritis. Possible significance of some recent observations. AB - Retroviruses have been proposed as etiologic agents for the development of chronic arthritis in humans. The arthritis seen in goats infected by caprine arthritis encephalitis virus and the spontaneous arthritis of inbred MRL/l mice illustrate how retroviruses may cause the development of a disorder closely resembling human rheumatoid arthritis. Several investigators have searched for evidence of retrovirus infection in patients with chronic arthritis, but in most cases the results have been disappointing. However, in 1983, Iversen isolated a virus-like particle from a patient with psoriasis. The particle had a buoyant density in sucrose and a protein composition that closely resembled murine and primate retroviruses. Particle proteins participate in immune complex formation in psoriasis, in psoriatic arthritis, and in ankylosing spondylitis. Particle proteins are also present in deposits in psoriatic lesions and in affected synovial tissue resembling immune complex deposits. The possible role for retrovirus-like antigens in the inflammatory process in psoriasis and seronegative arthritis is discussed. PMID- 2559477 TI - Use of intravenous immunoglobulins in Epstein-Barr virus induced thrombocytopenia. PMID- 2559479 TI - Graphite-furnace atomic absorption spectrometric determination of lead, cadmium, cobalt and nickel in infant formulas and evaporated milks after nitric-perchloric acid digestion and coprecipitation with ammonium pyrrolidine dithiocarbamate. AB - A graphite-furnace atomic absorption method, developed for lead and cadmium, was modified to enable simultaneous determination of lead, cadmium, cobalt and nickel in infant formulas and evaporated milks. The method was assessed on the basis of analytical quality assurance results during routine analysis of samples. Detection limits (ng g-1 based on 10 g sample size for ready-to-use formulas) were 0.04-0.21 for lead, 0.004-0.015 for cadmium, 0.04-0.2 for cobalt and 0.06 0.26 for nickel. Within-series repeatability and day-to-day reproducibility (among series) coefficients of variation (CVs) were, respectively, 11.7 and 16% for lead, 2.5 and 7.5% for cadmium, 7.2 and 18.8%for cobalt, and 8.1 and 8.6% for nickel at respective concentrations of 1.89, 0.40, 1.25 and 11.8 ng g-1. PMID- 2559480 TI - [Quantitative histoenzymological studies in the diagnosis of chronic viral hepatitis]. AB - The examination has involved 89 patients with chronic viral hepatitis, aged 14 to 59, 68 male and 21 female ones. Thirty-two (36.0%) patients suffered from chronic viral hepatitis B. Life-time biopsy of the liver was performed in all the cases. Chronic persistent hepatitis (CPH) has been diagnosed in 39 patients, chronic active one (CAH) in 50. Histochemical examinations of the biopsy specimens included measurements of alkaline phosphatase (AP) by the azocompound method, of 5-nucleotidase (5-Nuc) by the Ca-Co method, and of hepatocyte pigments. Stereologic methods were used in morphometric analysis of the area of sinusoids active for AP and 5-Nuc enzymes and of the intracellular pigment level with consideration for their distribution in the hepatic lobe. The findings evidence a significant increase of the area of sinusoids active for AP and 5-Nuc in CAH patients, in contrast to those with CPH, but the latter group has developed much higher levels of hepatocyte pigments. These data may be useful to specify the activity of chronic viral hepatitis in cases with poorly representative biopsy specimens. PMID- 2559481 TI - Surgical aspects of hepatic segmentation based on hepatic venographies. AB - Hepatic venographies were performed selectively in 42 patients with hepatocarcinoma. The findings were evaluated from anterior and lateral views. Thirty-nine right hepatic v. could be identified and the existence of one branch as the first ramification was found in 36 cases (92.3%). The first branches of the right hepatic v. could be classified into veins (V7) running from segment VII and those (V8) running from segment VIII. A V7 was identified in 26 cases (72.8%) and a V8 was identified in 10 cases (27.8%). The vena hepatica dorsalis (V8) running from segment VIII was recognised in 10 cases. The middle and left hepatic v. were identified in 31 cases and 33 cases respectively. Two main types of middle vein (one with no principal branching and the other with branching) were found in 11 cases (37.9%) and 12 cases (41.4%) respectively. The first branch of the middle hepatic v. (V8) running from segment VIII was identified in 10 cases (32.3%). These results indicate that anatomical consideration of the hepatic v. in each patient is necessary when performing hepatic resection. PMID- 2559482 TI - Cytomegalovirus retinitis. AB - Cytomegalovirus retinitis is a frequent and serious complication of various conditions, including diseases characterized by impaired immunity, such as the acquired immunodeficiency syndrome. Due to recent advances in the treatment of cytomegalovirus retinitis, accurate diagnosis and management have become increasingly important. In this review, the authors present the epidemiology, clinical features, diagnostic testing, treatment with complications, and prognosis of cytomegalovirus retinitis. A differential diagnosis is presented and the characteristic ocular lesions are illustrated. PMID- 2559483 TI - Apparent response of small cell lung cancer to an extract of mistletoe and homoeopathic treatment. AB - A patient with small cell lung carcinoma was treated initially with extracts of mistletoe and homoeopathic treatment and appeared to respond. Subsequently radiotherapy was given and the patient lived for five years seven months, which is much longer than is usual with this type of tumour. PMID- 2559484 TI - A double-blind randomized placebo controlled trial of thromboprophylaxis in major elective general surgery using once daily injections of a low molecular weight heparin fragment (Fragmin). AB - The safety and efficacy of the low molecular weight heparin fragment (Fragmin) administered as a single daily injection of 2,500 anti Xa units has been evaluated in 183 patients undergoing major elective general surgery. The study was double-blinded and placebo controlled. The active agent, or placebo, was given subcutaneously with the preoperative medication and continued postoperatively for 5-9 days. Ninety five patients received Fragmin and 88 were randomized to receive the placebo. The clinical characteristics of the two treatment groups were similar. Fragmin significantly reduced the incidence of deep venous thrombosis, as detected by a positive 125I fibrinogen leg scan, relative to the placebo treated patients (4/95, 4.2% v. 14/88, 15.9%; p = 0.008). The thrombotic events occurred predominantly (73%) amongst patients with malignancy. Haemorrhagic endpoints necessitating discontinuation of the trial treatment were 4% in each group. No severe adverse reactions or drug related deaths occurred. These results indicate that 2,500 anti Xa units of Fragmin given only once daily is effective thromboprophylaxis for patients undergoing major elective abdominal surgery. PMID- 2559485 TI - Thrombin-antithrombin III complexes in the prediction of deep vein thrombosis following total hip replacement. AB - In 196 consecutive patients who underwent elective total hip surgery we investigated the diagnostic accuracy of the thrombin-antithrombin III complex immunoassay, as assessed on the first, fourth and tenth postoperative day, for the development of deep vein thrombosis (DVT). Patients received either LMW heparinoid (n = 97) or placebo (n = 99) and underwent contrast venography on the tenth postoperative day. Thrombin-antithrombin III (T-AT) plasma levels were raised in all patients on the first postoperative day and gradually decreased during the study period. T-AT plasma levels were significantly higher in patients developing DVT when compared to patients without DVT and remained so until day 10. This difference was apparent both in the LMW-heparinoid group as well as in the placebo-treated patients. ROC-curve analysis revealed no satisfactory discriminative power for the diagnosis of developing DVT at any of the studied cut-off values for T-AT. We conclude that the postoperative determination of T-AT complex plasma concentrations in hip surgery patients has no clinical utility in the prediction of postoperative DVT. PMID- 2559486 TI - Effect of glycosaminoglycans on thrombin- and atroxin-induced fibrin assembly and structure. AB - This study was performed to quantitate the impact of several glycosaminoglycans (GAG) on fibrin assembly and structure. Gel formation was monitored as the increase in optical density at 633 nm subsequent to thrombin (2 NIH u/ml) or atroxin (0.10 mg/ml) addition to solutions of buffered fibrinogen (1 mg/ml) or plasma. Gel absorbance was measured as a function of wavelength (400 to 800 nm) and gel fiber diameter and mass/length ratio (mu) were calculated. Chondroitin sulfate A (CSA) shortened the lag phase, enhanced the maximal rate of turbidity increase, and increased the final gel turbidity of fibrin gels formed by thrombin or atroxin. CSA (16 mg/ml) increased fiber mu from 1.3 to 3.1 x 10(13) dalton/cm and fiber radius from 6.0 to 8.6 x 10(-6) cm in thrombin-induced gels. Mu increased from 0.7 to 2.7 x 10(13) dalton/cm and fiber radius from 4 to 7.8 x 10( 6) cm for atroxin-induced gels. Above 16 mg/ml, CSA caused fibrinogen precipitation in purified solutions but not in plasma. CSA inhibited thrombin induced plasma clotting of plasma but effects in atroxin-mediated plasma gels paralleled those seen in purified solutions. Chondroitin sulfate B (CSB)-induced changes in fibrin were similar but slightly less dramatic than those seen with CSA. Mu increased from 0.9 to 2.0 x 10(13) dalton/cm for atroxin-induced fibrin gels and from 0.8 to 2.3 x 10(13) dalton/cm for atroxin-induced gels. Low molecular weight heparin (Mr = 5100) slowed fibrin assembly and reduced fiber size by 50% in thrombin-induced gels. Changes in mu of atroxin-induced gels were much less pronounced (less than 20%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559488 TI - Purified human plasma kallikrein does not stimulate but primes neutrophils for superoxide production. AB - In patients with septicemia and septic shock the contact phase of blood coagulation is activated. It has been suggested that polymorphonuclear leukocytes (PMN) are directly activated by purified plasma kallikrein. This has been recently questioned because granulocytic elastase release induced by recalcification of normal and prekallikrein-deficient plasma was similar. We studied the interaction of different preparations of purified human plasma kallikrein with PMN. Cytosolic calcium shifts were measured with the quin2 method, PMN aggregation was assayed in an aggregometer, and superoxide production was quantitated as superoxide dismutase inhibitable cytochrome c reduction in a continuous assay. No increase of cytosolic free calcium was found during at least 5 min after adding 10 micrograms/ml plasma kallikrein to PMN. Similarly, highly purified plasma kallikrein from two different sources did not induce PMN aggregation at all, nor did it stimulate superoxide production. However, sequential exposure of PMN to plasma kallikrein and formylpeptide increased the superoxide production compared to stimulation with formylpeptide alone. This phenomenon which is called priming was observed at plasma kallikrein concentrations greater than or equal to 7 micrograms/ml. The active site of the molecule was required for the priming, because plasma prekallikrein, active site inactivated plasma kallikrein, and soybean trypsin inhibitor treated kallikrein did not prime PMN. This indicates that the contact activation system may play a role in host defence against bacterial infection. PMID- 2559487 TI - Eicosapentaenoic acid interferes with U46619-stimulated formation of inositol phosphates in washed rabbit platelets. AB - Eicosapentaenoic acid (EPA) inhibits platelet responsiveness to aggregating agents. To investigate the reactions that are affected by EPA, we examined the effect of preincubating aspirin-treated rabbit platelets with EPA on stimulation of inositol phosphate formation in response to the TXA2 analogue U46619. Stimulation of platelets with U46619 (0.5 microM) caused aggregation and slight release of dense granule contents; aggregation and release were inhibited by preincubation of the platelets with EPA (50 microM) for 1 h followed by washing to remove unincorporated EPA. Incubation with EPA (50 microM) for 1 h did not cause a detectable increase in the amount of EPA in the platelet phospholipids. When platelets were prelabelled with [3H]inositol, stimulation with U46619 of control platelets that had not been incubated with EPA significantly increased the labelling of inositol phosphates. The increases in inositol phosphate labelling due to U46619 at 10 and 60 s were partially inhibited by preincubation of the platelets with 50 microM EPA. Since the activity of cyclo-oxygenase was blocked with aspirin, inhibition of inositol phosphate labelling in response to U46619 indicates either that there may be inhibition of signal transduction without a detectable change in the amount of EPA in platelet phospholipids, that changes in signal transduction require only minute changes in the fatty acid composition of membrane phospholipids, or that after a 1 h incubation with EPA, activation of phospholipase C is affected by a mechanism that is not directly related to incorporation of EPA. PMID- 2559489 TI - Time dependent factor IX inhibitor. PMID- 2559490 TI - Induction of tissue factor-like activity of human monoblastic leukemia cell line by tumor necrosis factor-alpha. AB - Cell lysates of the human monoblastic leukemia cell line, THP-1, have procoagulant activity (PCA) that is Ca++-dependent and not demonstrable in either Factor VII-, or Factor X-deficient plasma. The PCA of THP-1 cells was enhanced by human recombinant tumor necrosis factor-alpha (TNF-alpha) up to five fold. There was a dose-dependent increase in PCA when THP-1 cells were cultured with concentrations of TNF-alpha, up to 10 U/ml. PCA of cell lysates or whole cell preparations was measured in comparison to a rabbit brain thromboplastin standard. The effect of TNF-alpha was enhanced by recombinant human interferon gamma (IFN-gamma). Cycloheximide inhibited the induction of PCA by THP-1 cells, which shows that the protein synthesis is essential to mediate the effect of TNF alpha. THP-1 cells and U937 cells bound 125I-labeled TNF specifically. The numbers of receptors per cell were found to be 1,890 and 1,550 for THP-1 and U937 cells, respectively. Other lymphoid and myeloblastic leukemia cell lines examined did not have TNF receptors, indicating that the effect of TNF-alpha is mediated by the receptors on the cell surface. PMID- 2559491 TI - Antiplatelet effects of protopine isolated from Corydalis tubers. AB - Protopine inhibited the aggregation and ATP release of rabbit platelets induced by ADP, arachidonic acid, PAF, collagen and ionophore A23187. Although the platelet aggregation caused by thrombin was not inhibited by protopine (100 micrograms/ml), the release reaction was partially suppressed. In rabbit platelet rich plasma, protopine also inhibited the platelet aggregation caused by ADP, arachidonic acid, PAF and collagen. The thromboxane B2 formation of washed platelets caused by arachidonic acid, collagen, ionophore A23187 and thrombin was suppressed by protopine. Protopine inhibited the intracellular calcium increase caused by arachidonic acid in quin-2/AM loaded rabbit platelets. In the presence of indomethacin, the intracellular calcium increase caused by collagen and PAF was completely suppressed by protopine, and the intracellular calcium increase caused by thrombin was partially inhibited. The phosphoinositides breakdown caused by collagen and PAF was inhibited by protopine, but that by thrombin was not affected significantly. Protopine did not cause the elevation of cyclic AMP level of platelets. It is concluded that the antiplatelet effects of protopine is due to inhibition on thromboxane formation and phosphoinositides breakdown and then lead to the decrease of intracellular calcium concentration. PMID- 2559492 TI - Pharmacology of a potent, new antithrombotic agent, 1,3-dihydro-7,8-dimethyl-2H imidazo[4,5-b]quinolin-2-one (BMY-20844). AB - The effects of 1,3-dihydro-7,8-dimethyl-2H-imidazo[4,5-b]quinolin-2-one (BMY 20844) on platelet function and experimental thrombosis were evaluated in a series of in vitro, ex vivo and in vivo experiments. The compound inhibited platelet aggregation in vitro in platelet rich plasma obtained from humans, rats and rabbits with EC50s of less than 1 microgram/ml when aggregation was induced by ADP, collagen or thrombin. Supra-additive interaction against ADP aggregation was also observed when BMY-20844 was combined with prostacyclin. BMY-20844 was orally active with an ex vivo ED50 in the rat of 3.2 mg/kg vs ADP. Significant antithrombotic activity was observed in two animal models (laser induced thrombosis in the microcirculation of the rabbit ear and coronary artery thrombosis in the dog). Inhibitions of 52% at 3 mg/kg p.o. in the laser model and 100% at 1 mg/kg i.d. in the coronary artery thrombosis model were obtained. Modest inotropic and hemodynamic effects were observed in ferrets and dogs. BMY 20844 was found to be a potent, specific inhibitor of platelet low Km cyclic AMP phosphodiesterase. PMID- 2559494 TI - [Parvovirus infections in dogs and cats: problems of immunization]. AB - The general problems of immunization against parvoviroses are only understandable when knowing the basic principles of epidemiology, pathogenesis and immunology of those virus infections. The main difficulties with regard to vaccinations are 1. efficacy of homologous respectively heterologous vaccines, 2. indications for live resp. inactivated vaccines, 3. combination with other vaccines, 4. bridging of the "immunological gap" (refractory phase) of puppies, 5. mode of application and timing of vaccinations (schedule of vaccinations), 6. failure in vaccinated animals, 7. complications originating from vaccinations, 8. passive immunization. PMID- 2559493 TI - Exogenous glycosaminoglycans modulate chondrogenesis, cyclic AMP level and cell growth in limb bud mesenchyme cultures. AB - Effects of hyaluronate, heparin and chondroitin-6-sulfate were studied on micromass cultures of chick limb bud mesenchyme (Hamburger and Hamilton stages 23 24). Histochemical, electron microscopical, biochemical and radiochemical investigations of day 4 cultures revealed dose-dependent inhibitory effects of these glycosaminoglycans on chondrogenesis, cyclic AMP level and growth of cells. In addition, hyaluronate with 100 micrograms/ml dose caused a displacement of newly formed proteoglycan from cultures into the medium. It is supposed that exogenous glycosaminoglycans influence ionic equilibrium in the immediate vicinity of cells and disturb the organization of the prechondrogenic extracellular matrix resulting in alterations of cell membrane--cytoskeleton associations. These alterations may provoke a reduction in cyclic AMP level and DNA synthesis. It is suggested that a reduction in cyclic AMP level preceding the expression of cartilage phenotype results in the inhibition of chondrogenesis. PMID- 2559495 TI - [Poxvirus infection in a cat]. AB - For the first time, a poxvirus infection was diagnosed as an etiologic agent of dermal disease in a living domestic cat in Germany. A literature survey, the clinical symptoms of the infection and the diagnostic procedures are described. Poxvirus infections should be considered as a differential diagnosis in feline dermatologic problems. PMID- 2559496 TI - [A new herpesvirus-caused disease in tortoises]. AB - This report describes a highly fatal diphtheroid-necrotizing stomatitis in tortoises of hitherto unknown etiology. The tortoises suffered from dyspnea and anorexia, due to massive diphtheroid membranes in oral and pharyngeal cavities. Histologically, eosinophilic intranuclear inclusion bodies were found in epithelial layers of oral and tracheal mucous membranes. Furthermore, electron microscopy revealed herpesvirus like particles in affected cells. Possible environmental factors influencing the outbreak of the disease are discussed. PMID- 2559497 TI - Mechanism of mammalian ovulation. AB - The sequence of events within the ovary during the process of ovulation discussed in this review is schematically represented in Fig. 1. It is obvious that LH, perhaps with some contribution from FSH, is the normal physiological trigger for the ovulatory sequence of events, and it appears from the available information that the effects of LH are mainly mediated via adenylate cyclase and increased cAMP levels. The cAMP in turn, via cAMP-dependent protein kinase, influences at least three distinct steps in the ovulatory process which seem to be of crucial importance, namely 1) the stimulation of steroidogenesis; 2) the stimulation of cyclooxygenase/lipooxygenase leading to increased prostaglandin/leukotriene synthesis; and 3) the stimulation of plasminogen activator which catalyzes the conversion of plasminogen to plasmin. A fourth crucial step in the ovulatory mechanism is the LH-induced increase in latent collagenase, but it remains to be determined if this step is mediated via cAMP. Concomitant with the increase in latent collagenase, there also appears to be an LH-dependent increase in collagenase inhibitors. The latent collagenase is then activated, and it appears that leukotrienes and prostaglandins, as well as plasmin, may be involved in this process. The active collagenase causes a digestion of the collagen in the follicle wall, and plasmin, as well as possibly other proteolytic enzymes such as proteoglycanases, may cause a further dissociation of the follicular wall. These processes of digestion of collagen and dissociation of the collagen fibers result in an opening in the follicular wall with the formation of the stigma and rupture. While the weakening of the follicular wall takes place throughout the entire wall, rupture remains for the most part a localized process at the apex of the follicle. This localization of the rupture may be explained on the basis of mechanical factors operating when the follicle wall thins and weakens. While it is clear that prostaglandins and leukotrienes can influence smooth muscle by causing contractions and that these compounds can cause vascular changes such as increased permeability, vasodilation, and vasoconstriction, it is not clear what the exact role of these latter processes are in ovulation. It appears that progesterone and not estrogen play an important role in the mechanism of LH induced follicular rupture, but the locus of action of progesterone and its mechanism of action remains to be determined.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2559498 TI - Connective tissue breakdown in ovulation. AB - A meshwork of collagen over the apical region of the follicle must be breached to permit the ovum to escape. We propose that specific collagenase activity is responsible for collagen breakdown in this region. Immature rats are primed with pregnant mare serum gonadotropin (PMSG), followed at 48 h by hCG. At 8 h after hCG, collagenase activity, measured in extracts of ovarian tissue, is elevated about five-fold. Ovulation follows at 10-12 h. Ovaries from PMSG-primed rats are dissected at 48 h, placed in a perfusion apparatus, and perfused with luteinizing hormone and 3-isobutyl-1-methyl xanthine. The ovulations induced by this treatment can be blocked to the extent of 70% with a synthetic collagenase inhibitor. The activation of procollagenase is believed to involve plasminogen activator and plasmin. In support of this, we find that tranexamic acid at 1 mM inhibits ovulation about 70%. The inhibitor must be added within 3-4 h of LH to be effective. A specific plasmin inhibitor, D-Val-Phe-Lys-chloromethyl ketone, is similarly effective. PMID- 2559499 TI - The role of ovarian proteases and their inhibitors in ovulation. AB - To assess the role of inhibitors of proteolytic enzymes, such as plasminogen activator (PA) and collagenase in the ovulatory process, inhibitor activity and mRNA levels were examined in periovulatory rat and human ovaries. In the rat, immature animals received 20 IU of pregnant mare serum gonadotropin (PMSG) followed 52 h later by 10 IU of hCG. Ovaries were removed at intervals from 0 to 20 h after human chorionic gonadotropin (hCG) administration. Inhibitor activity for metalloproteinases, such as collagenase, increased from 60.5 +/- 4.1 inhibitor units/ovary at 0 h (i.e., time of hCG treatment) to a maximum of 218.2 +/- 11.4 units/ovary at 8 h after hCG before decreasing at 12 h (time of ovulation) and 20 h (122.2 +/- 7.9 and 71.6 +/- 8.1 units/ovary, respectively). Human follicular fluid and granulosa cells were obtained from preovulatory follicles of patients in our in vitro fertilization program. Metalloproteinase inhibitor activity was evaluated in follicular fluid as well as the levels of PA and PA inhibitor (PAI) mRNA by Northern analysis. Increasing metalloproteinase inhibitor activity was positively correlated with follicular levels of estradiol (p less than 0.001) and progesterone (p less than 0.02, N = 26). Chromatographic separation of follicular fluid resulted in two peaks of metalloproteinase inhibitor activity. The large molecular weight (MW) inhibitor had an approximate size of 700 kilodaltons (kDa) and may represent alpha 2-macroglobulin, a serum derived inhibitor. The small MW inhibitor shared many of the characteristics of tissue-derived inhibitors of metalloproteinases. Partial purification of the small MW inhibitor by Concanavalin A-Sepharose and Heparin-Sepharose chromatography demonstrated the inhibitor to be a glycoprotein with an approximate MW = 28-29 K. Northern analysis of human granulosa cell total RNA from preovulatory follicles showed little or no detectable tissue-type PA or urokinase-type PA mRNA. In contrast, two species of PA inhibitor type-1 mRNA were detected in relative abundance. The present findings demonstrate the presence of proteolytic inhibitors in periovulatory ovaries of the rat and human. These ovarian inhibitors may play a role in regulating connective tissue remodeling during follicular rupture. PMID- 2559500 TI - Adenosine as substrate and receptor agonist in the ovary. AB - In the present study the possible dual effects of adenosine as substrate and adenosine receptor agonist in rat granulosa cells, cumulus-oocyte complexes, luteal cells and ovarian membranes are discussed. Adenosine is an indispensable compound in cell energy metabolism, as precursor to cofactors, second messenger and nucleic acids. Adenosine is also an agonist to adenosine receptors. The adenosine receptor can either inhibit (A1) or stimulate (A2) adenylate cyclase. Alternatively, in some cells adenosine receptor activation is linked to other cellular events like inhibition of Ca2+ fluxes. Adenosine is taken up by isolated preovulatory granulosa and luteal cells from pregnant mare serum gonadotropin treated immature rats, but follicle stimulating hormone (FSH) decreases the uptake by granulosa cells. Adenosine, but not the non-metabolizable adenosine analogs 5'-(N-ethyl)carboxamide-adenosine (NECA), 2-chloro-adenosine (2-Clado), N6-(R-phenyl-isopropyl)-adenosine (R-PLA) and N6-(S-phenyl-isopropyl)-adenosine (S-PLA), increase granulosa cell ATP levels. FSH and luteinizing hormone (LH) decrease granulosa cell ATP levels in the presence or absence of adenosine. It has previously been shown that FSH and LH decrease oxygen consumption by cumulus oocyte complexes and increase their lactate production. These effects have been suggested to be due to a competition of cofactors (e.g. ADP) common to glycolysis and the respiratory chain. The fact that adenosine reverse the gonadotropin induced effects on oxygen consumption and lactate production support this theory. Adenosine and its analogs increase cAMP accumulation in luteal and granulosa cells only in the presence of gonadotropins, and this effect is antagonized by the adenosine receptor antagonist 8-phenyl-theophylline (8-PHT). Furthermore, adenylate cyclase is stimulated by adenosine analogs in membranes from non luteinized and luteinized ovarian membranes and in luteal cell homogenates. The effect of NECA is antagonized by 8-PHT. In the membranes, the rank order of potency was NECA greater than 2-Clado greater than R-PLA greater than S-PLA, suggesting adenosine A2 receptors. In summary, it is suggested that adenosine can act both as a substrate to intracellular metabolism and as an adenosine A2 receptor agonist in granulosa and luteal cells. A paracrine short loop positive feedback model is proposed where extracellular adenosine, derived from a gonadotropin-induced extracellular increase in cAMP and a decrease in cellular ATP, enhances gonadotropin stimulation in granulosa and luteal cells. PMID- 2559501 TI - Modulation of porcine granulosa cell functions by interleukin-1. AB - Increasing evidence suggests that functions of the immune system and gonads are closely related with each other. In cultures of granulosa and luteal cells, macrophages have been shown to modulate steroidogenic functions. In this paper we present the modulatory effects of interleukin-1, a cytokine produced predominantly by activated macrophages, on gonadotropin-induced differentiation, as well as growth of cultured porcine granulosa cells. PMID- 2559502 TI - [The diagnosis of intravital mechanical trauma by the blood EPR-spectral parameters]. AB - The author studied the possibility of diagnosing the lifetime occurrence of mechanical trauma by electron paramagnetic resonance on the basis of values of a constant related to the rate of reaction of spin probe recovery by blood in different time periods up to 28 days of postmortem interval. PMID- 2559503 TI - Neuroendocrine-immune interactions in T cell ontogeny. PMID- 2559504 TI - In vitro activity of nalidixic acid and its iron (III) complex on Entamoeba histolytica. AB - The in vitro activity of the antibacterial agent nalidixic acid (HNal) and its iron (III) complex (FeNal) against Entamoeba histolytica HM1 strain trophozoites in axenic or monoxenic (associated with Clostridium symbiosum) cultures was investigated. Using a dilution test with TYI-S-33 medium, this protozoan was found to be susceptible to both drugs, but FeNal showed amoebicidal activity only at concentrations higher than those used with HNal. PMID- 2559505 TI - Kinetics of the immune responses during the course of hepatic amoebic infection- an experimental study. AB - Following intrahepatic inoculation of axenic Entamoeba histolytica (strain NIH:200V), all hamsters developed amoebic abscesses in the liver by 7 d after inoculation. All the animals cleared the infection spontaneously by 28 d after inoculation. The establishment of amoebae leading to acute liver abscess was accompanied by significantly reduced levels of antibodies to the plasma membrane (PM) protein of E. histolytica, reduced direct macrophage cytotoxicity, and reduced anti-PM antibody-mediated macrophage-dependent cytotoxicity to amoebic trophozoites. This phase of infection was also accompanied by a significant reduction in cellular sensitization to PM proteins and to phytohaemagglutinin. Reduction in size of the liver abscesses and clearance of infection were accompanied by significant development of anti-PM antibodies, cellular sensitization to the PM proteins, enhanced direct macrophage cytotoxicity, and anti-PM antibody-mediated macrophage-dependent cytotoxicity to trophozoites. The data suggest that the amoebic disease process is modulated by immune responses to the PM protein of E. histolytica. PMID- 2559507 TI - High susceptibility of five axenic Entamoeba histolytica strains to gossypol. AB - The antiamoebic potency of gossypol was tested against 5 axenic Entamoeba histolytica strains, in logarithmic phase growth in PEHPS medium. All of the strains were moderately susceptible to this polyphenolic drug. The 50% inhibitory concentration (IC50) was of the same order of magnitude in all strains: 0.015 microM (for strain HM-1) to 0.067 microM (for HM-38). The difference between the IC50 of HM-1 and the remaining 4 strains (HM-38, HK-9, HM-2 and HM-3) was significant, although it was greater between HM-1 and HM-38 (P less than 0.005) than between HM-1 and the other 3 strains (P less than 0.05), for which the IC50 was 0.03 microM, 0.049 microM and 0.38 microM respectively. Gossypol is more toxic in vitro for amoebae than other drugs widely used clinically, its pharmacological effects and safe dosage in humans are well known because of its antifertilizing effect, and it is accumulated mainly in the liver and colon. Accordingly, this compound could be a good antiamoebic agent if it is as potent in vivo as it is in vitro. PMID- 2559508 TI - Pathogenic zymodemes of Entamoeba histolytica in Japanese male homosexual population. PMID- 2559506 TI - Role of rotavirus and enteric adenovirus in acute paediatric diarrhoea at an urban hospital in Mexico. AB - Of 150 acute diarrhoea paediatric cases admitted to the General Hospital Augustin O'Horan in Merida, Yucatan, Mexico between January 1986 and December 1986, human rotavirus was detected in the faeces of 48 (32%) either as the sole aetiological agent or in association with other viruses. Adenovirus was detected in faeces of 18 (12%) patients. Rotavirus of the long pattern type (antigenic subgroup II) predominated. Children aged 7 to 12 months were most commonly infected. Rotavirus was detected throughout the year, whereas adenovirus was detected between January and September only. Clinical features of patients with rotavirus and adenovirus infections are compared. PMID- 2559509 TI - Lack of an association between acute gastroenteritis, acute respiratory infections and malaria in young Gambian children. AB - The incidence of acute gastrointestinal and acute respiratory infections was measured in 2 groups of approximately 750 Gambian children aged 3-59 months during a 3-year period. One group of children was partially protected against malaria by fortnightly chemoprophylaxis with Maloprim whilst children in the other group were infected much more frequently. Mortality from acute gastroenteritis and from acute respiratory infections was similar in the 2 groups. The proportions of children in each group who complained of gastrointestinal or severe respiratory symptoms on morbidity surveillance were also similar. Thus, no evidence was found to suggest that malaria plays either a direct or indirect role in causing acute gastrointestinal or respiratory infections in young children in The Gambia. PMID- 2559511 TI - Concentration of Pentostam in human breast milk. PMID- 2559510 TI - Virulence of pathogenic and non-pathogenic zymodemes of Entamoeba histolytica (Indian strains) in guinea-pigs. AB - Guinea-pigs were inoculated with Entamoeba histolytica strains isolated from cases of amoebic liver abscess, amoebic dysentery and asymptomatic cyst passers and the strains were classified in zymodemes by isoenzyme electrophoresis. It was observed that certain non-pathogenic zymodemes were potentially pathogenic for the guinea-pigs. The relationship of zymodemes and virulence in laboratory animals is discussed. PMID- 2559512 TI - Epidemiology and aetiology of acute childhood diarrhoea in Burma: a rural community survey. AB - Identical epidemiological and microbiological surveys were carried out in a rural community at Intakaw, Burma, in 2 months, one in the hot wet and the other in the cool dry season. The incidence of diarrhoea was highest in children under 3 years old and in the hot wet month when enterotoxigenic Escherichia coli was the predominant pathogen. Rotavirus was the commonest pathogen in the cool dry month but was absent in the hot wet month. Shigellae, salmonellae and campylobacter were isolated in both seasons, but were not significantly commoner in patients than in controls. Vibrios and Yersinia enterocolitica were sought but not found in any specimen. PMID- 2559513 TI - HIV1 and HIV2: two ancient viruses for a new disease? PMID- 2559514 TI - Arbovirus infections and viral haemorrhagic fevers in Uganda: a serological survey in Karamoja district, 1984. AB - Sera collected in May 1984 from 132 adult residents of Karamoja district, Uganda, were examined by haemagglutination inhibition tests for antibodies against selected arboviruses, namely Chikungunya and Semliki Forest alphaviruses (Togaviridae); dengue type 2, Wesselsbron, West Nile, yellow fever and Zika flaviviruses (Flaviviridae); Bunyamwera, Ilesha and Tahyna bunyaviruses (Bunyaviridae); and Sicilian sandfly fever phlebovirus (Bunyaviridae); and by immunofluorescence tests against certain haemorrhagic fever viruses, Lassa fever arenavirus (Arenaviridae), Ebola-Sudan, Ebola-Zaire and Marburg filoviruses (Filoviridae), Crimean-Congo haemorrhagic fever nairovirus and Rift Valley fever phlebovirus (Bunyaviridae). Antibodies against Chikungunya virus were the most prevalent (47%), followed by flavivirus antibodies (16%), which were probably due mainly to West Nile virus. No evidence of yellow fever or dengue virus circulation was observed. A few individuals had antibodies against Crimean-Congo haemorrhagic fever, Lassa, Ebola and Marburg viruses, suggesting that these viruses all circulate in the area. PMID- 2559515 TI - Ainhum. PMID- 2559516 TI - Genetic approaches to the determination of structure-function relationships of G protein-coupled receptors. AB - The beta-adrenergic receptor (beta AR), which has been extensively characterized pharmacologically, serves as a useful model system for the analysis of the structure-function relationships of G protein-coupled receptors. Genetic and biochemical analysis has revealed that the ligand binding domain of the receptor involves residues within the hydrophobic transmembrane core of the protein. Molecular substitution experiments suggest that adrenergic agonists and antagonists are anchored to the receptor through an ionic interaction between Asp113 in the third hydrophobic region of the receptor and the protonated amine group of the ligand. In addition, catecholamine agonists are bound through hydrogen bonding interactions between two serine residues in the fifth hydrophobic domain of the receptor and the catechol hydroxyl groups of the ligand. Agonist-mediated activation of the G protein Gs requires residues within the cytoplasmic loop linking the fifth and sixth transmembrane helices which are predicted to form amphipathic alpha-helices. The strong structural similarities among G protein-coupled receptors imply that the information gained from genetic analysis of the beta AR should be applicable to other hormone and neurotransmitter receptors of this class. PMID- 2559517 TI - New advances in molecular pharmacology of Ca2+ channels. PMID- 2559518 TI - GABAB receptors and their significance in mammalian pharmacology. AB - Much progress has been made in GABAB receptor pharmacology since the discovery of this receptor in 1980. Selective agonists and antagonists have been developed and a functional role for the receptor as a mediator of slow inhibitory postsynaptic potentials in many brain regions has emerged. In this article, Norman Bowery discusses the evidence for heterogeneity of GABAB receptors, their possible physiological and pathological roles and the therapeutic potential of GABAB receptor agonists and antagonists. PMID- 2559519 TI - Multiplicity of GABAA--benzodiazepine receptors. AB - Binding studies suggest the presence of at least two pharmacologically distinct 'central' benzodiazepine receptors in the brain. Since central benzodiazepine receptors are allosteric modulatory sites on GABAA receptors, this evidence indirectly points to the existence of at least two GABAA receptors. Werner Sieghart describes biochemical studies that have identified several different alpha- and beta-subunits of these receptors, and molecular biological studies in which the genes encoding a variety of different alpha-, beta- and gamma-subunits have been isolated, sequenced and expressed in Xenopus oocytes. These studies all point to the existence of multiple GABAA receptors in the brain. PMID- 2559520 TI - The GABAA receptor: molecular biology reveals a complex picture. PMID- 2559522 TI - Activity of rifabutin alone and in combination with clofazimine, kanamycin and ethambutol against Mycobacterium intracellulare infections in mice. AB - Treatment of mice for 12 weeks with clofazimine or kanamycin decreased the number of organisms from lungs, liver and spleen of mice infected with Mycobacterium intracellulare N-260, compared with findings seen with rifabutin or ethambutol. Treatment with various drug combinations (rifabutin-ethambutol, rifabutin clofazimine, rifabutin-kanamycin, rifabutin-ethambutol-clofazimine, rifabutin ethambutol-kanamycin, and rifabutin-clofazimine-kanamycin), particularly in the presence of clofazimine, enhanced elimination of the organisms from these organs. PMID- 2559521 TI - Coagglutination test: a simple and rapid diagnostic technique for goat pox. AB - The coagglutination test was standardised using Staphylococcus aureus strain Cowan I (containing Protein A) coated with anti-goat pox serum for the detection of goat pox antigen of infected goat skin or kid kidney cell culture antigen. Agglutination was observed within 10 seconds in virus dilutions up to 10(-6). As the test is easy to perform it can be used for rapid diagnosis of goat pox. PMID- 2559523 TI - Creatine kinase-BB activity in malignant tumors and in sera from patients with malignant diseases. AB - Creatine kinase (CK EC 2.7.3.2) and CK-BB activity was analyzed in 41 malignant tumors of 6 different sites and different histological structures. The same analyses were done on 150 sera of patients with malignant diseases of various localizations. The rate of CK activity was determined kinetically, whereas tissue and serum CK-BB were separated chromatographically (Mercer). Insofar as malignant tumor tissues are concerned, the highest average rate of CK-BB activity was detected in tumors of the prostate (mean 1450 IU/g), and the lowest in tumors of the parotid gland (mean 5.2 IU/g). CK-BB was detected by the Mercer technique in 56 (37.3%) of 150 analyzed sera of patients with malignant diseases. The rate of CK activity in sera of patients with malignant diseases was 8 to 74 IU/I. In comparison with the site of the malignant process no significant CK serum activity differences were observed. T2-T3 tumors did not significantly influence the activity of either CK or CK-BB in the case of either tissues or sera (T1-T3). Enzyme activity was found to be much higher--both in tumoral tissue and in sera- with T4 tumors. The highest rate of CK-BB activity was found in sera of patients with malignant tumors of the stomach (mean 8.1 IU/I), and the lowest in malignant tumors of the rectum (mean 1.8 IU/l). PMID- 2559524 TI - In vivo and in vitro growth of SCLC cells derived from biopsies. AB - In order to increase the availability of SCLC cells derived from biopsies, in vivo and in vitro growth methods were investigated. The cells grown in both conditions were periodically monitored for reactivity with 2 monoclonal antibodies (MAbs): MLuC1 directed against SCLC cells and IM1 which recognizes the class II antigen on activated lymphocytes and macrophages. About 50% of the 28 analyzed SCLC specimens were found to proliferate in one or both systems. The in vitro-grown cells exhibited the same heterogeneity found in the original cell suspensions and moreover, in some cases only normal cells were recovered after several in vitro passages. From the subcutaneous transplanted tumors a large number of MLuC1-positive tumor cells could easily be recovered, thus indicating the validity of the in vivo methodology. The MBr1 MAb, directed against an epithelial antigen, was found to react with about 50% of the 26 tested tumors, mainly those which demonstrated in vivo and/or in vitro growth capacity. These data suggest that only some tumors, presumably with peculiar biological characteristics, can efficiently grow in these artificial systems. PMID- 2559525 TI - Bone marrow biopsy in the staging of small cell lung cancer. AB - From April 1982 to December 1987, 71 patients with small cell lung cancer entered a randomized clinical trial, and underwent bone marrow biopsy (BMB) as part of staging procedures. We identified 8 patients (11%) with bone marrow metastases, 6 with extensive disease independently of BMB, and 2 with extensive disease on the basis of the BMB only. BMB determined a change in the stage in only 3% (2/71) of the cases. No differences were found in the hematological parameters of the patients with or without bone marrow metastases. The median survival of the patients with bone marrow involvement was the same (41 weeks) as those with extensive disease but without bone marrow involvement. We conclude that unilateral BMB without aspiration detects a substantial proportion of bone marrow metastases in patients with extensive disease. This fact does not worsen the prognosis. A small proportion of patients with apparently limited disease has bone marrow involvement. The technique therefore contributes, to a small extent, to the definition of the clinical stage of the disease. However, bone marrow involvement is an important data of natural history, and therefore new methods to better assess this peculiar site of the disease are needed. PMID- 2559527 TI - Glioblastoma multiforme of the cerebellum in an elderly man. A case report. AB - Glioblastoma multiforme of the cerebellum is rare and comprises a small fraction of all glioblastomas. Eight-five cases have been reported in the literature to date. A 75 year old man is reported with a left cerebellar glioblastoma multiforme. The pathogenesis, course, treatment and prognosis are reviewed. PMID- 2559526 TI - Large cell neuroendocrine carcinoma of the lung. AB - Large cell neuroendocrine (LCNE) carcinomas of the lung are a newly recognized, highly aggressive and frequently misdiagnosed entity. We report a case of stage I LCNE lung carcinoma initially misdiagnosed as large cell undifferentiated carcinoma or poorly differentiated adenocarcinoma. The tumor was very extensively necrotic and its neuroendocrine differentiation was only demonstrable with immunohistochemical staining with PHE-5 monoclonal antibody and with antisera against synaptophysin and calcitonin. ACTH, somatostatin and neurofilaments were not demonstrable. The clinical course was ominous and the patient died within 17 months. The reason for this rapid fatal outcome could be ascribed either to the neuroendocrine phenotype of the tumor, or to the extensive necrosis, or both. PMID- 2559528 TI - Adrenal failure due to bilateral metastases as the sole manifestation of relapsing lung carcinoma. Report of two cases. AB - We describe two patients with lung carcinoma in whom adrenal glands were the sole site of tumor relapse, revealed by the appearance of Addison's disease. Both patients showed bilateral adrenal masses on US and/or CAT scans and received hormone replacement therapy, with rapid improvement of their general conditions. One of them, with small-cell carcinoma, could also be treated with further chemotherapy and achieved a second remission. Therefore, we stress that patients with lung carcinoma should be periodically screened for adrenal deposits by US or CAT and undergo prophylactic steroid maintenance whenever metastatic involvement of the glands is detected. PMID- 2559529 TI - [Surgical treatment of benign hepatoma]. AB - The authors share their personal experience with treatment of 28 patients with benign tumors of the liver. The method of choice in treatment of this contingent of patients in thought to be resection of the liver. Radical operations were performed in 23 patients. Good nearest and remote results were followed up during the period from 1 year to 17 years. PMID- 2559531 TI - Experimental caprine retrovirus infection in sheep. PMID- 2559530 TI - Studies of the effect of recombinant human-alpha 1 interferon on experimental parainfluenza type 3 virus infections of the respiratory tract of calves. AB - The effect of the administration of recombinant human interferon on the severity of clinical disease and the extent of pneumonic lesions in calves infected experimentally with bovine parainfluenza 3 (PI3) virus was studied in two experiments. In the first, three pairs of calves aged seven to 10 days were used; one of each pair was injected intramuscularly with 10(6) units of interferon/kg bodyweight for three consecutive days, and the other was left untreated. On the day after the first injection of interferon all the calves were challenged with PI3 virus, a different dose being administered to each pair. There was no evidence of any protective effect from the treatment with interferon. The second experiment used eight, six-week-old calves; four were inoculated in the same way with interferon and all the calves were challenged with the same dose of PI3. Again, there was no evidence of a reduction either in the severity of clinical disease or in the extent of lung consolidation in the calves treated with interferon. PMID- 2559532 TI - Equine influenza. PMID- 2559533 TI - Investigation of "mysterious" disease in livestock: hydrocyanic acid poisoning. AB - An investigation of "mysterious" disease due to hydrocyanic acid (HCN) poisoning in livestock in this state was carried out. Detailed clinicopathological and pathological studies were conducted. Characteristic signs of acute tympany followed with profuse frothing, convulsions and dyspnea were recorded. Cynosis of the mucosa with characteristic anoxemic tissue changes and a high concentration of HCN in rumen content, feed and skeletal muscles were recorded. These were sufficient to establish the diagnosis. Successful treatment with a specific antidote was achieved, and further morbidity and mortality was checked. PMID- 2559534 TI - Changes in B cell and T cell subsets in bovine leukaemia virus-infected cattle. AB - Direct immunofluorescence and fluorescence-activated cell sorter techniques were used for the detection of surface immunoglobulin positive (SIg+) cells in peripheral blood lymphocytes (PBL's) of bovine leukaemia virus (BLV) infected cattle with or without persistent lymphocytosis (PL+, PL-) and in BLV-free cattle. The percentage of SIg+ cells was more than twice as high in BLV+PL+ cattle than in BLV-free and BLV+PL- cattle. Bovine T cells, and T cell subsets were identified indirectly by the same techniques using three monoclonal antibodies (MAb's) specific for all T cells (IL-A43), T helper (BoT4) cells (IL A12) and T cytotoxic (BoT8) cells (IL-A17). The major histocompatibility complex (MHC) determinants of both class II (BoT4) and class I (BoT8) as well as all T cells were significantly reduced in BLV+PL+ compared to BLV-free cattle. The actual decrease in the BoT8 cell subset or the dilution effect that would change effector:target cell ratio suggests that a resultant decrease in cytotoxic activity in BLV+PL+ cattle may play an important role in the progress of BLV infection in cattle. PMID- 2559535 TI - Immunological responses of cross-bred and in-bred miniature pigs to swine poxvirus. AB - Swine poxvirus (SPV), topically and subdermally applied to skin of the inguinal region of cross-bred and in-bred miniature pigs, caused typical pox lesions to occur with a pustular stage at 4 to 5 days p.i., and healing by 10 to 14 days p.i. Following inoculation, peripheral blood lymphocytes (PBLs) of the pigs showed lower transformation responses to SPV and mitogens (Concanavalin A, phytohemagglutinin and 12-0-tetradecanoyl-phorbol 13-acetate) than PBLs from uninoculated controls. The PBLs generally responded to SPV from 7 to 9 days p.i. to 23 to 30 days p.i. with a maximum transformation response at the 12 to 13 days p.i. interval. Sera from the animals generally showed presence of SPV neutralizing antibody as early as 7 days p.i. and a peak titer at 20 days p.i. of 1:512. No detectable SPV-antibody was observed at 50 days p.i. By 51Cr release assays, PBLs displayed the ability to lyse target cells in the presence of SPV antibody with peak lysis from the 11th through the 21st day p.i. An antibody histocompatibility restricted cell lysis was observed at 11 and 14 days p.i. When PBLs were depleted of adherent cells, there was a reduction in lysis of target cells indicating the adherent cells were instrumental as effector cells in the presence of SPV-antibody. Control pigs not exposed to SPV showed no PBL response to SPV-antigen. Partially histocompatible and non-histocompatible porcine kidney cells were found useful as cell models for evaluating SPV-infected pigs in their effort to immunologically respond to SPV. PMID- 2559536 TI - Observations on blood leucocytes and lymphocyte subsets in sheep infected with bovine leukaemia virus: a progressive study. AB - Haematological parameters and reactivity of lymphocyte antigens to monoclonal antibodies were studied over a 10-month period in sheep experimentally infected with bovine leukaemia virus (BLV). BLV-inoculated animals seroconverted within 1 month and showed a significant lymphocytosis 2-6 weeks after infection. Control animals inoculated with BLV-free lymphocytes showed a stronger and more immediate neutrophil response than those inoculated with BLV-positive lymphocytes. One month after infection, BLV-inoculated sheep showed a relative increase of cells bearing antigens T4, T6, T8 and T19, and 10 months into the trial, MHC II lymphocytes increased, T6 remained elevated, but T4 helper cells were significantly decreased in number. Lymphoma tissue showed the presence of T8 cells, and lymph nodes from seroconverted sheep had areas of concentrated T4 staining cells. These results demonstrate responses in cellular immune mechanisms to infection with BLV. PMID- 2559537 TI - Cross-neutralizing and subclass characteristics of antibody from horses with equine infectious anemia virus. AB - Antibody responses in horses with equine infectious anemia virus (EIAV) were examined to determine their cross-neutralizing capacity. Antibodies induced by infection with any of six biologically cloned variants of EIAV cross-neutralized multiple variants from the group. Anti-EIAV antibody was found in both the IgG and IgG(T) subclasses in plasmas with virus-neutralizing activity and the majority of antiviral antibody was of the IgG(T) subclass. Depletion of IgG(T) did not increase the neutralization indexes of either neutralizing or non neutralizing plasma samples. PMID- 2559538 TI - Bovine recombinant interleukin-2 augments immunity and resistance to bovine herpesvirus infection. AB - The in vivo administration of bovine recombinant interleukin-2 (rIL-2) was evaluated in calves vaccinated and then challenged with bovine herpesvirus-1 (BHV 1). In Experiment 1, 24 calves were allotted to four groups: control; bovine rIL 2; BHV-1 vaccine (modified-live); and bovine rIL-2 + BHV-1 vaccine. Serum neutralizing antibody titers to BHV-1 were increased sixfold, and virus shedding was fourfold less in calves vaccinated and treated with rIL-2 (25 micrograms/kg, intramuscularly) when compared to calves that received vaccine only. Treatment with rIL-2 induced lymphokine-activated killer activity that was eliminated by pretreating effector cells with complement and a monoclonal antibody (B26A) specific for the sheep red blood cell receptor. The rIL-2 treatment in BHV-1 vaccinated calves increased the calves' ability to withstand a BHV-1 challenge. However, during treatment with rIL-2, calves developed diarrhea and mild fever that abated after IL-2 treatment was stopped. A second experiment was then conducted to determine a dose of rIL-2 that would enhance immunity to BHV-1 without causing adverse side effects. Twenty-five calves were allotted to five groups that received injections of rIL-2 at 0.0, 25.0, 2.5, 0.25, or 0.025 micrograms kg-1 day-1 for 5 days. All calves received a modified-live BHV-1 vaccine. Calves treated with 25.0 micrograms kg-1 day-1 showed similar adverse side effects as in the first experiment but all other calves were normal. Compared to control calves, those treated with 25.0, 2.5, and 0.25 micrograms kg 1 day-1 of rIL-2 had higher (P less than 0.05) serum antibody titers to BHV-1 and following challenge lower (P less than 0.05) BHV-1 titers in nasal secretions; additionally, clinical disease as evidenced by nasal and ocular discharge was less severe (P less than 0.05). In vitro cytotoxic responses against BHV-1 infected bovine kidney cells were increased (P less than 0.05) in calves treated with rIL-2 in a dose dependent manner. These data suggest that bovine rIL-2 at 2.5 to 0.25 micrograms/kg may be an effective adjuvant to immunization. PMID- 2559539 TI - Polymorphonuclear leucocyte function: relationship between induced migration into the bovine mammary gland and in vitro cell activity. AB - Low doses of 10(-7) mg Escherichia coli endotoxin applied as intramammary infusion into single bovine quarters induced a rise in milk cell count without other inflammatory signs. Significantly fewer quarters responded in early lactation than in mid lactation. Maximum cell count was also somewhat later and less pronounced in early lactation. The rise in milk cell count after infusion of E. coli endotoxin was related to in vitro chemotactic activity of blood polymorphonuclear leucocytes (PMN). PMN isolated from cows which did not respond with a rise in milk cell count upon endotoxin infusion showed a diminished chemotactic activity in vitro as compared to PMN isolated from animals which did respond to an intramammary endotoxin infusion with a rise in milk cell count. No differences in phagocytic and metabolic activity were observed in vitro between the PMN isolated from the two groups of animals. PMID- 2559540 TI - Comparison of the sequence of the secretory glycoprotein A (gA) gene in Md5 and BC-1 strains of Marek's disease virus type 1. AB - DNA fragments containing the secretory glycoprotein A (gA) gene of Marek's disease virus type 1 (MDV1) were cloned from the DNA libraries of very virulent Md5 and virulent BC-1 strains and sequenced. Two open reading frames (ORF1 and ORF2) were identified for both strains. The ORF1 has the potential to code for a protein of 501 amino acids with a molecular weight of 56 kD that contains strong hydrophobic regions in both the amino and carboxyl termini, and nine potential N linked glycosylation sites, while the ORF2 is capable of coding for a 24-kD protein. These results indicate that the ORF1 codes for the unprocessed form of gA. Between the Md5 and BC-1 strains, only two sequence mismatches exist in the DNA fragment. More differences appear to exist in the gA sequence of the MDV1 GA strain (12), which lacks a strong hydrophobic anchor sequence. Similarities between the predicted amino acid sequences of the MDV1 gA and the proteins of the other herpesviruses such as herpes simplex type I gC, pseudorabies virus gIII, and varicella zoster virus gpV were noted. PMID- 2559541 TI - Characterization of human T-cell lines harboring defective human immunodeficiency virus type 1. AB - Defective HIV-producing T-cell lines were subcloned from MT-4/HIVHTLV-IIIB' MOLT 4/HIVHTLV-IIIB, and H9/HIVHTLV-IIIB cell lines chronically infected with HIV. The NY-M10 cell line derived from MOLT-4/HIVHTLV-IIIB and the NY-H6 cell line derived from H9/HIVHTLV-IIIB produce defective HIV, which lacks the ability to infect human T-cell lines. NY-M10 cells retain the capacity to form multinucleated giant cells in cocultivation with HIV-uninfected CD4-positive cells. However, NY-H6 cells failed to fuse with CD4-positive cells. Electron microscopic analysis indicated that the defective HIV produced from NY-M10, like those reported previously, lacked the structure of the nucleocapsid, and the virion released from NY-H6 was indistinguishable from those of authentic HIV particles. Southern and Northern blotting analyses of NY-M10 and NY-H6 cleared that the genome of those defective viruses was not significantly deleted, suggesting minor mutation(s) should take place on the viral genome. PMID- 2559542 TI - A region of basic amino-acid cluster in HIV-1 Tat protein is essential for trans acting activity and nucleolar localization. AB - The trans-acting factor of human immunodeficiency virus (HIV), Tat, has a basic amino-acid cluster that is highly conserved among different HIV isolates. We have examined the effects of mutations in the basic region of Tat on its trans-acting activity and cellular localization. Introduction of a stop codon immediately preceding the basic region abolished the activity, while the truncated mutant with the basic region retained some activity. The basic region of Tat was replaceable with that of Rev (another trans-acting factor of HIV) but not with that of adenovirus Ela nor cellular enzyme. The result of immunofluorescence analysis revealed a correlation between the nuclear, especially nucleolar, accumulation and the activities of mutant Tat proteins. PMID- 2559543 TI - [The effect of cytomegalovirus and Epstein-Barr virus infection on immunologic signs]. AB - Patients with active CMV or EBV infection characterized by the presence of specific serum IgM and/or high immunofluorescence in IgG, have higher levels of circulating immune complexes and a lower phagocytic activity of leucocytes than patients with latent CMV/EBV infections or not infected patients. In active infections at the same time the absolute number of lymphocytes with the surface sign CD8 increases. The amount of lymphocytes with differential antigens CD3 and CD4 does not change substantially. Also the levels of the third and fourth component of complement and C-reactive protein remain within the normal range. PMID- 2559544 TI - [The effect of Sinecal fiber on compensation in type 2 diabetics]. AB - The authors investigated in 28 type 2 diabetics the effect of addition of 21 g Sinecal fibre to the normal diet on the glucose and lipid metabolism. The value of C,peptide was significantly influenced after breakfast and a single dose of Sinecal. The other investigated parameters (blood sugar on fasting and after a meal, glycosuria, glycosylated proteins, C-peptide on fasting and after a meal, triacylglycerol and cholesterol) were not influenced by a single nor by long-term intake of Sinecal. Further development of fibre preparations with better tolerance and a greater effect on the glucose metabolism is desirable. PMID- 2559546 TI - [ATPase activity, cyclic nucleotide levels in the heart and lipid peroxidation in the aorta of spontaneously hypertensive rats]. AB - Activities of Na+,K+- and Ca2+, Mg2+- ATPases as well as content of cAMP were increased in heart tissues of spontaneously hypertensive rats as compared with normotensive animals. On the other hand, the rate of lipid peroxidation was decreased in aorta of the hypertensive rats. PMID- 2559545 TI - [Changes in the cAMP levels and acid phosphatase activity in a monolayer primary culture of hepatocytes from newborn rats during anoxia and substrate deprivation]. AB - Acid phosphatase activity and cAMP level were studied in primary hepatocyte culture of new born rats under conditions of anoxia and substrate deprivation (incubation of the cells in Hanks salt solution). Incubation of hepatocytes in Hanks salt solution within one hour under conditions of anoxia caused a significant increase in free (cytosolic) enzyme activity. Substitution of Hanks salt solution by normal tissue culture medium and reoxygenation after 1 hr anoxia resulted in a decrease of free acid phosphatase activity, whereas activity of the enzyme in lysosomal fraction was increased. Content of cAMP was decreased distinctly after 15 min incubation of hepatocyte culture under conditions of anoxia and substrate deprivation and was increased above control values within 5 min of reoxygenation and substitution of Hanks salt solution by normal tissue culture media. Addition of cAMP-containing liposomes to hepatocyte culture under these experimental conditions led to a decrease in free acid phosphatase activity and to an increase of the enzyme activity in lysosomal fraction. cAMP appears to modulate the lability of hepatocyte lysosomal membrane. The mechanisms involved in these processes are discussed. PMID- 2559547 TI - [The effect of alloxan on activity of various enzymes of phosphorus metabolism in vitro]. AB - Effect of alloxan on activity of acetylphosphatase, alkaline and acid phosphatases, phenylpyrophosphatase was studied in vitro. Alloxan at concentrations 100 microM-10 mM inhibited acid phosphatase and especially alkaline phosphatase. Reducing agents (pyrocatechol, hydroquinone, ascorbic acid), which are easily converted into corresponding oxidized forms, at concentrations 100 microM-1.0 mM activated alkaline phosphatase in rat kidney and small intestine. PMID- 2559548 TI - Development of an immunoassay for the detection of minute amounts of IgG-coated erythrocytes in whole blood and its application for the assessment of Fc-mediated clearance of anti-D-coated erythrocytes in vivo. AB - A rapid and sensitive immunoassay for the detection of minute quantities of IgG coated erythrocytes in whole blood was developed. Washed red blood cells were incubated in two steps with anti-human IgG antiserum followed by 125I-labelled protein A. The assay was able to detect amounts of sensitized erythrocytes as small as 0.5 ml of packed erythrocytes in a total blood volume of 5 liters and hematocrit 40%. A linear relation between increasing amounts of IgG-coated red cells in whole blood and the binding of 125I-labelled protein A was obtained. We applied the technique on the assessment of the removal of IgG anti-D-coated erythrocytes from the circulation of test individuals. T1/2 for the elimination of approximately 4 ml packed red cells sensitized with 62 micrograms of anti-D in 14 normal subjects was 20 +/- 5 min (mean +/- SEM). A splenectomized person did not clear the injected cells from the circulation during the test period of 70 min. If a standard curve was constructed the total blood volume in the test subjects could be calculated. This value correlated well (r = 0.99) with the blood volume calculated from the height and weight of the test individuals. PMID- 2559549 TI - [Barrett's esophagus]. AB - In 12 patients the endoscopic examination raised the suspicion of Barrett's syndrome. By means of guided biopsies and static scintigraphy with gamma camera and 99m-technetium pertechnetate ectopic gastric mucosa was found in the esophagus of 5 of the patients. In one patient a Barrett's ulcer was found. The establishment of a focus of increased radioactivity proximal from the stomach by static scintigraphy is a proof for the presence of ectopic gastric mucosa in the esophagus. Because of the high risk of malignancy a systemic endoscopic biopsy control is necessary for the patients with proved columnar-cell metaplasia in the esophagus. PMID- 2559551 TI - Human papilloma virus (HPV) and uterine cervix cancer. PMID- 2559550 TI - [Cardiac metastasis in primary cancer of the liver]. AB - A rare case of a 77-year-old man with primary liver cancer and an isolated metastasis in the right atrial endocardium is presented. The metastasis was 30/40 mm large and occupied a considerable part of the atrial cavity. Clinically the disease was manifested by progressing chronic right ventricular failure, ending with a total heart failure. The diagnosis was made at the post mortem examination. The rarity of the case is pointed out. The possibility of a correct diagnosis while the patient was still alive is discussed. PMID- 2559552 TI - Further studies on the oxidative cleavage of the pentyl side-chain of cannabinoids: identification of new biotransformation pathways in the metabolism of 3'-hydroxy-delta-9-tetrahydrocannabinol by the mouse. AB - 1. Oxidative degradation of the pentyl side-chain of cannabinoids leading to compounds containing even numbers of carbon atoms was studied by investigating the in vivo metabolism of 2'- and 3'-hydroxy-delta-9-tetrahydrocannabinol (THC). 2. The hydroxy cannabinoids were administered i.p. to mice, the livers were removed after 1 h and extracted metabolites were identified by g.l.c.-mass spectrometry. 3. The major metabolic route for both compounds was hydroxylation at the allylic 11-position followed by oxidation to a carboxylic acid. Additional hydroxylation occurred at C-8. 4. Little oxidative degradation of the side-chain was found for 2'-hydroxy-delta-9-THC but abundant metabolites were formed by this route from the 3'-hydroxy compound. 5. The major metabolites of this type were acids containing three carbon atoms in the chain. These are the normal products of beta-oxidation but their formation from an (omega-2)-hydroxy intermediate appears novel. 6. Other metabolites contained two carbon atoms in the side-chain but were alcohols rather than acids and were again apparently formed by novel mechanisms. 7. The results indicate that the major route leading to cannabinoid metabolites with two-carbon side-chains (loss of three carbon atoms) is initiated by omega-2 hydroxylation. PMID- 2559553 TI - [Rapid diagnosis in Clostridium perfringens wound infections]. AB - Prompt and early microbiological differential diagnosis is essential for clinical presumptive diagnosis of gas gangrene. The differential diagnosis includes clostridial myositis (gas gangrene), clostridial cellulitis and other gas producing infections. Examination of Gram preparation (bacterioscopy) and detection of the etiologic agent in muscle specimens are necessary for diagnosis. Clostridium perfringens has been shown as the causative organism of gas gangrene. A method is reported which allows the screening and identification of Clostridium perfringens from clinical specimens in a few hours. Using a medium yielding optimal growth and toxin production, pure cultures are centrifuged and subjected to rapid tests (detection of beta-galactosidase, phospholipase C). PMID- 2559554 TI - Endothelial protection by defibrotide--a new strategy for treatment of myocardial infarction? AB - Myocardial ischemia is associated with endothelial injury and an apparently insufficient generation of endothelium-derived vasodilating and platelet and white cell inhibitory mediators, such as prostacyclin (PGI2) and EDRF. This paper reviews some recent findings of our laboratory on cardioprotective effects of defibrotide, a PGI2 stimulating agent, in experimental myocardial ischemia and its possible sites of action in several in vitro assay systems. Defibrotide (32 mg/kg x h) reduced the infarct size by 50% in pigs, subjected to 1 h of coronary artery ligation followed by 3 h of reperfusion. This was associated with significant inhibition of neutrophil activation during the reperfusion period and a two-to threefold increase in cardiocoronary PGI2 generation. In vitro studies on PAF- and calcium ionophore-stimulated human granulocytes confirmed a dose dependent (10-1000 micrograms/ml) antineutrophil effect of defibrotide (inhibition of lysosomal enzyme release) which was independent of the type of stimulus. Defibrotide (0.1 mg/ml) also inhibited superoxide anion generation from PAF stimulated neutrophils in Langendorff-perfused guinea pig hearts and was equipotent to a specific PAF antagonist (BN 52021). Defibrotide (0.1 mg/ml) did not stimulate PGI2 release from cultured porcine aortic endothelial cells but enhanced PGI2 release four- to fivefold above control if endothelial cells were coincubated with platelets. These data demonstrate a considerable cardioprotective potential of defibrotide which appears to involve endothelial protection from granulocyte-derived noxious compounds and a long-lasting stimulation of PGI2 production. PMID- 2559555 TI - [Clinical and electroneurographic results following surgical treatment of the ulnar nerve sulcus syndrome]. AB - The sulcus of the elbow-joint represents a bottleneck which may cause compression of the ulnar nerve. We studied clinical and electroneurographical results in 26 patients after operation of the sulcus ulnaris-syndrome. The influence of the duration of compression as well as additional illnesses was evaluated on the postoperative result. The surgical treatment was an anterior transposition of the nerve in a subcutaneous layer combined with external neurolysis. Due to the symptoms and the electroneurographical results after operation the patients were divided into three groups. The best results were obtained in cases with a short period of preoperative complaints (below 1 year). Although heavy work had a positive effect on the manifestation of a sulcus ulnaris-syndrome, it had no adverse effect if started again after operation. Anterior subcutaneous transposition is the operative procedure of choice in cases of sulcus ulnaris syndrome. PMID- 2559556 TI - Metabolic regulation through second-site phosphorylation. AB - Protein phosphorylation is the most prevalent covalent reversible modification used for the regulation of many biological functions. The dynamic phosphorylation state of cellular proteins results from the balance between the activities of protein kinases and phosphatases. Even autophosphorylation, shared by practically all protein kinases, is also considered as a mode of enzyme regulation. Phosphodephosphorylation reactions seem also to be regulated at the substrate level, and the structure of a target protein can be strongly altered through phosphorylation in a way which may either promote or depress the further action of a protein kinase or phosphatase. Second-site phosphorylation may have various consequences on the kinetic properties of the enzyme-substrate. Glycogen synthase is possibly one of the most complex examples of enzyme regulation through interactions between phosphorylation sites. An interesting example of regulation at the substrate level in a network protein phosphorylations, is provided by the effects of casein kinase-1 (CK-1), casein kinase-2 (CK-2) and protein kinase FA (PKFA) on the modulator protein of the ATP, Mg-dependent phosphatase. The crucial step in the activation of the enzyme is the transient phosphorylation of the modulator at the Thr-72 residue catalyzed by PKFA. The CK-2 mediated phosphorylation occurs at three Ser residues: Ser-86, Ser-120 and Ser-121. Although by itself CK-2 does not elicit any phosphatase activity, it potentiates the subsequent activation brought about by PKFA. The phosphorylation mediated by CK-1, probably directed toward Ser-86, blocks the activation of the ATP,Mg dependent phosphatase mediated by PKFA. PMID- 2559557 TI - [Retroviruses. Problems in electron microscopic diagnosis]. AB - A department of electron microscopy with specialisation in virus diagnosis evaluates the results concerning retrovirus findings. In the course of research work already classified retroviruses were observed, numerous unexpected retrovirus diagnoses being of special interest. This concerned exclusively to ultra-thin sections of permanent and immune cells. Expounded in this paper are principles by which examiners should abide in any case. Reference is also made to the importance of individual findings. PMID- 2559558 TI - [Syringomas of the vulva]. AB - A case of a 23-year-old female patient showing syringomas on the labia majora pudendi is reported. Clinical and histological findings are described, the differential diagnosis is discussed. PMID- 2559559 TI - Investigations on the influence of copper succinate on the production of superoxide anion radicals by bovine small intestinal mucosa cells. AB - The effect of an experimental ischemia lasting for 45 minutes and a subsequent period of reperfusion of equal length on the activity of xanthine oxidase (XO) and microsomal NADPH-cytochrome P450 reductase (NADPH-CR) were investigated in the small intestinal mucosa of male neonatal calves of the breed German "Schwarzbunte". The activity of the NADPH-CR was determined by chemiluminescence. The activity of XO decreased during ischemia, but rose to values above the control level following reperfusion. 5 mg of Cu2(succinate)2 (CuSu) administered either intraarterially or intraluminally during reperfusion prevented the rise in XO. Formation of malondialdehyde decreased in the presence of CuSu. The NADPH-CR likewise showed subnormal activity values during ischemia, but also remained at a low level during reperfusion. The activity of this enzyme was further lowered by local intraarterial or intraluminal administration of 5 mg of CuSu and by 120 mg of CuSu administered intravenously during the reperfusion. These results are discussed with regard to the superoxide anion radical induced tissue lesions observed during reperfusion. PMID- 2559560 TI - [Simian AIDS virus: review]. AB - The review article on Simian AIDS viruses compiles the presently known facts of AIDS-related simian retroviruses causing immunodeficiencies or lymphomas in nonhuman primates. They are also compared to the situation in man. Simian acquired immunodeficiencies or lymphoproliferative diseases can be caused by infectious (exogenous) oncoviruses (C- or D-type) or by lentiviruses. Infectious D-type oncoviruses of simian origin are: Mason-Pfizer Monkey Virus (MPMV), Simian Retrovirus type 1 (SRV-1), Simian Retrovirus type 2 (SRV-2). The simian D-type oncoviruses do not have a counterpart in man. Infectious C-type oncovirus of monkeys is STLV, related to, but not identical with HTLV. Most serious to man might be simian lentivirus-infections (SIVs) due to the genetic instability of many lentiviruses. The properties of the different viruses, the clinical symptoms and morphological lesions caused by these agents are outlined and the possible dangers to man discussed. MPMV and the SRVs are considered as genetically stabilized and probably not infectious to man, at least no D-type virus infections of man are known. STLV should be treated with caution due to the close genetic relationship of man and nonhuman primates and of STLV and HTLV, although even in STLV-positive colonies no human infections were reported so far. Extreme caution seems advisable regarding the SIVs, not only because of the close phylogenetic and genetic relationships to HIVs. HIVs and SIVs, which are considered to have diverged only comparably recently from common ancestors and from each other possess a marked genetic plasticity (unstability). Therefore working with Old World monkeys, especially of African origin, with tissues of blood obtained from such animals requires certain protective action. PMID- 2559562 TI - Seroprevalence of porcine and human influenza A virus antibodies in pigs between 1986 and 1988 in Hassia. AB - 1,268 sera collected from slaughtered pigs in Hassia (FRG) from 1986 to 1988 were tested for antibodies against porcine and human influenza A virus strains using the single radial haemolysis test (SRHT). Antibodies against the porcine strains (subtype H1N1) A/Swine/Arnsberg/1/81, A/Swine/Iowa/15/30 and A/New Jersey/7/76 were detected in 411 (32.4%), 318 (25.1%) and 304 (24.0%) of sera, respectively. Up to 1988 a slight increase (10%) in the seroprevalence to A/Swine/Arnsberg/1/81 was noticed, whereas the results obtained with the other strains showed little variation. Antibodies against the human H1N1 strain A/Singapore/6/86 were only found in sera collected 1987 and 1988 in rates of 1.6% and 3.0%. Serological indication of infections with the human H3N2 strains A/Victoria/1/75, A/Hong Kong/1/68 and A/Philippines/2/82 could be shown in 286 (22.6%), 178 (14.4%) and 135 (10.6%) of the serum samples. Within the three year period the rate of sera positive for antibodies against A/Philippines/2/82 increased from 6.5% to 23.0%, whereas no variation in the rates were found using the other H3N2 strains. Antibodies simultaneously against porcine (H1N1) and human (H3N2) virus strains were detected in 9.9% of all sera tested. PMID- 2559561 TI - [The detection of bovine herpesvirus type 1 (BHV 1) using an intradermal test. I. Field studies]. AB - An intradermal test (delayed hypersensitivity test) for the diagnosis of BHV1 infection was evaluated in 791 cattle of 16 dairy farms. The skin reactions were compared with the results of serological examinations using a commercial BHV1 ELISA kit (Trachitest). As antigen concentrated, purified and inactivated BHV1 was used. The skin reaction (increase of the skin fold thickness) was used for the interpretation of test results. The best results were obtained with the control of the skin reaction on the third day after injection of the antigen. From 393 serologically BHV1 negative cattle with an age of more than 6 months 391 (99.5%) had a skin reaction up to 1.0 mm and 2 animals (0.5%) had a reaction of 1.3 and 1.9 mm, respectively. The mean increase of skin fold thickness was 0.2 mm. Out of 291 serologically BHV1 positive cattle with an age of more than 6 months 270 had antibodies from natural infection and, partially, from additional vaccination with inactivated BHV1 vaccine. 266 (98.5%) of these animals showed a skin reaction of more than 2.0 mm, in 3 animals (1.1%) a skin reaction up to 1.0 mm was observed and 1 animal (0.4%) had a reaction of 2.0 mm. The mean increase of the skin fold thickness was 6.3 mm. 21 animals had BHV1 antibodies only because of vaccination with inactivated BHV1 vaccine. Only 4 animals had a skin reaction of more than 2.0 mm. Among 107 animals with an age up to 6 months 30 were serologically BHV1 positive and 77 were BHV1 negative. In all animals the skin reaction was less than 1.0 mm, the mean was 0.2 mm. PMID- 2559563 TI - Equine coital exanthema (EHV-3 virus) infection in India. AB - A progenital disease encountered at one equine stud farm at Bangalore in Southern India during 1987 was investigated and confirmed as equine coital exanthema on the basis of characteristic lesions and clinical symptoms, isolation of equine herpes virus-3 (EHV-3) from the scabs collected from animals having active lesions and demonstration of neutralizing antibodies in the sera of recovered mares and stallion. This is the first authenticated report of the occurrence of equine coital exanthema in India due to EHV-3. PMID- 2559564 TI - [An ultracytochemical study of the hydrogenosomes of Trichomonas vaginalis]. AB - The ultracytochemical study of cultural T. vaginalis has been carried out. The activity of magnesium-dependent ATPase, transport (sodium-potassium-dependent) ATPase and adenylate cyclase was detected in dense corpuscles, located on the membranes of hydrogenosomes having a rounded form. The authors suggest that the dense corpuscles are the zones of the active transport of ions through the membranes of hydrogenosomes. PMID- 2559565 TI - Characteristics of MNNG induced repair synthesis and DNA synthesis induced by human cytomegalovirus. AB - DNA synthesis induced by N-methyl-N'-nitro-N-nitrosoguanidine in mouse embryo and human embryo cells was compared with DNA synthesis induced in these cells by human cytomegalovirus. In virus infected human embryo cells grown in the medium depleted of arginine DNA synthesis showed resistance to hydroxyurea and arabinofuranosylcytosine, similarly as repair synthesis induced by MNNG. DNA synthesis induced by the virus in mouse embryo cells was partially sensitive to both inhibitors. PMID- 2559566 TI - [Cystosarcoma phyllodes of the breast]. PMID- 2559567 TI - Spontaneous retroperitoneal haematoma presenting as femoral neuropathy. Case report. AB - A 16-year-old boy presented with a sudden onset of unilateral femoral neuropathy. This was caused by a spontaneous retroperitoneal haematoma. Surgical decompression produced a dramatic improvement in the patient's condition. PMID- 2559568 TI - [Fechtner syndrome: report of two families and review of the literature on the related disorders]. AB - Two families who had been reported as having May-Hegglin anomaly were diagnosed as having Fechtner syndrome based on complication of renal disease, deafness and characteristic leukocyte inclusions. By Wright-Giemsa staining, the inclusions of neutrophils were smaller and stained less clearly than those seen in May-Hegglin anomaly. Ultrastructurally, the inclusions consisted of organelle-poor cytoplasm, containing small particles, which were probably ribosomes. Some inclusions contained only a few filaments or small remnants of rough endoplasmic reticulum (RER). The latter resembled the findings observed in Fechtner syndrome. They lacked the parallel arrays of filaments characteristic of May-Hegglin anomaly and the parallel strands of RER seen in toxic Dohle bodies. Renal disorders ranged from microscopic hematuria to renal failure requiring dialysis in one family and persistent proteinuria in the other family. Deafness was sensorineural type or combined type due to otitis media. Except for the fact that they lacked congenital cataracta, the findings in these two families were consistent with those in Fechtner syndrome. PMID- 2559569 TI - [Aplastic crisis due to human parvovirus B19 infection in glucose-6-phosphate dehydrogenase deficiency]. AB - Human parvovirus B19 is known to cause aplastic crisis in patients with hemolytic anemias due to cytotoxic effect of the infection to erythroid progenitor cells. We report here the first case of aplastic crisis by B19 in a patient with glucose 6-phosphate dehydrogenase deficiency. A five-year-old boy was admitted to the hospital because of severe anemia, fever and jaundice. Four weeks after admission, he developed erythema infectiosum. B19 infection was confirmed using countercurrent immunoelectrophoresis, Southern blotting and hybridization method, and radioimmunoassay for B19 specific IgM. B19 virus antigen was detected by an indirect immunofluorescent method in both the cytoplasm and nucleus of large mononuclear cells that had no granules in bone marrow. On admission, the hemoglobin was 3.1 g/dl and no reticulocytes were detected in the peripheral blood smear. Bone marrow examination revealed a normocellular marrow with erythroid hypoplasia and M/E ratio of 7.38. Large basophilic erythroblasts containing vacuoles were also noticed. Elevation of indirect bilirubin and hemoglobinuria suggested intravascular hemolysis. Transient mild thrombocytopenia associated with increased PAIgG was observed. It is likely that B19 virus infection caused hemolysis which contributed to severe anemia. PMID- 2559570 TI - The POEMS syndrome among Chinese: association with Castleman's disease and some immunological abnormalities. AB - POEMS or Crow-Fukase syndrome is a multisystemic, clinically malignant disorder of obscure etiology. Peripheral neuropathy and plasma cell dyscrasia are central features. The authors now report 7 Chinese patients with this syndrome in which PCD or paraproteinemia were absent in 6, and 2 had a lymph node histology resembling that of hyaline-vascular Castleman's disease. Immunological abnormalities consisted of either increased or decreased numbers of B- and T cells in 2 cases, and an elevated OKT4/OKT8 ratio with paradoxical dissociation of the lymphocyte transformations to various concentrations and types of mitogens in 1 case. This suggests that the underlying abnormalities of POEMS syndrome are heterogeneous and that it may be an immunologically related syndrome of varying etiology. PMID- 2559571 TI - Biphasic recovery in n-hexane polyneuropathy. A clinical and electrophysiological study. AB - We studied the course of recovery in n-hexane polyneuropathy in 4 patients by quantitatively assessing clinical and electrophysiological features. The electrophysiological study included measures of motor conduction of the median, ulnar, tibial and peroneal nerves and sensory conduction of the median, ulnar and sural nerves. After cessation of exposure, there was an initial worsening in muscle strength, sensory deficit and nerve conduction for up to 2-5 months. This deterioration was more severe and prolonged in the lower limbs than in the upper limbs. The period of deterioration was followed by a slow recovery studied for 1 year. PMID- 2559572 TI - Activity distribution of cytochrome oxidase in the rat retina. A quantitative histochemical study. AB - Cytochrome oxidase (CYO) is a key enzyme in the respiratory chain. Therefore, CYO has an important role in the cell metabolism. In the present study CYO activity in the rat retina was identified by histochemical staining. The density of the staining, corresponding to the activity of the CYO, was evaluated quantitatively by densitometry. A high CYO activity was found in the retinal pigment epithelium, in the inner segment of the photoreceptors, in the outer plexiform layer and in the inner plexiform layer. In the outer segment of the photoreceptors, in the outer nuclear layer, and in the inner nuclear layer the CYO activity was relatively low. An analysis of variance demonstrated that the precision in an estimation of a mean depends on the number of animals and the number of retinal sections per animal. PMID- 2559573 TI - The focal treatment of retinoblastoma with emphasis on xenon arc photocoagulation. AB - Two-hundred-seventy-eight retinoblastoma tumors in 169 patients were photocoagulated by one of two doctors with the xenon arc photocoagulator. More than 70% of tumors treated were cured by photocoagulation. The following features correlated with success: 1) size of the tumor, 2) location of the tumor and 3) elevation of the tumor. Seventy of 72 tumors (97%) up to 1 dd in size were cured while only 9 of 22 (41%) tumors larger than 5 dd were cured. The mean size of tumors cured was 2 dd, while the mean size of those that failed was 3.9 dd. Tumors located anterior to the equator were more often successfully treated (67/81 = 83%) than those posterior to the equator (44/74 = 60%). Tumors with the height equal to half the base were classified as 'low elevation', while those with heights larger than half the base diameter were classified as high elevation. Tumors with low elevation did significantly better (162/201 = 81%) than those with high elevation (33/76 = 43%). No correlation was found between success and the following features: age at diagnosis, sex, age at first photocoagulation, eye involved (right or left), time from radiation to first light coagulation, nasal versus temporal tumors or the clock hour of the tumor or category of tumor independent of size of location. Forty-four percent of tumors that failed photocoagulation went on to develop vitreous seeding and 55% required enucleation. Fifty percent of the eyes that were treated initially with photocoagulation went on to develop new tumor foci elsewhere in the eye. In all cases the new tumors appeared anterior to the equator. The children who developed additional tumors in the eye were younger (5.5 months) when photocoagulated than those who did not develop additional tumors (47.75 months). Photocoagulated retinoblastomas must be followed for at least three years before a cure is certain. PMID- 2559574 TI - [Temporal bone metastasis of adenocarcinoma of the breast: personal observations and a review of the literature]. AB - A single case of temporal bone metastases from breast carcinoma is reported and its clinical features and main instrumental diagnostic characteristics are described. Although this is quite rare the possibility cannot be neglected when making diagnosis should the clinical features and test results suggest such an expansive process of the temporal bone. In fact, although extremely limited, treatment is strictly linked to proper and early diagnosis. PMID- 2559575 TI - Natural killing of varicella-zoster virus (VZV)-infected fibroblasts in normal children, children with VZV infections, and children with Hodgkin's disease. AB - We studied mononuclear cell (MNC)-mediated natural killing (NK) of varicella zoster virus (VZV)-infected fibroblasts in normal children, children with VZV infections, and children with Hodgkin's disease. NK activity was tested in 18 hr 51Cr release assays. NK activity for adults was significantly higher than that for children 1-3 years old or 4-6 years old (p less than 0.05). Serological status did not affect NK activity. NK activity in normal children was not increased 4-6 weeks after immunization with varicella vaccine. Seven normal children with natural varicella showed significantly higher NK activity against VZV-infected and uninfected targets. Eight immunosuppressed children with herpes zoster showed significantly reduced NK activity within 72 hours of the onset of herpes zoster. However, their NK activity rose to the normal level one to two weeks later. Children with Hodgkin's disease had low NK activity. These results suggested that NK cells might play an initial defensive role in VZV infections, and that low NK activity in immunocompromised hosts might contribute to their high incidence of herpes zoster. PMID- 2559576 TI - Prevalence of cytomegalovirus antibodies in Brazilian women of childbearing age and newborns. PMID- 2559577 TI - [Synthesis and biological activity of derivatives of guanosine 3', 5'-cyclic phosphate]. AB - Methyl, n-butyl 2'-TBDMS-N2-DMF-guanosine 3',5'-cyclic phosphate and 2'-TBDMS-N2 DMF-guanosine 3',5'-cyclic diethyl-phosphoramidate were synthesized by reaction of protected guanosine with trivalent phosphorus reagents in the presence of tetrazole followed by oxidation. The reaction occurred stereospecifically. Protected guanosine 3',5'-cyclic phosphotriesters and N, N'-diethyl phosphoramidate were shown to have inhibitory activity on the synthesis of DNA and RNA in mouse liver tumor cell. Diastereoisomers of n-butyl N2-substituted guanosine 3',5'-cyclic phosphate have been shown to activate adenylate cyclase in vitro. PMID- 2559578 TI - [Effects of zinc sulfate on isolated vascular smooth muscles]. AB - ZnSO4 was found to inhibit the contractions of isolated guinea pig aortic preparations induced by KCl, CaCl2 and noradrenaline (NE). ZnSO4 (0.01-0.1 mmol/L) shifted the dose-response curves for KCl, CaCl2 and NE to the right, and depressed their maximal responses with PD2' values of 3.21, 3.07 and 2.91 respectively. ZnSO4 (0.1 mmol/L) significantly antagonized KCl, CaCl2 and NE induced contraction responses in isolated dog portal vein preparations. These results suggest that the ZnSO4 induced relaxation of vascular smooth muscle might result from its antagonistic effect on transmembrane Ca2+ channels. PMID- 2559579 TI - [A facile synthesis of adenosine 3', 5'-cyclic methylphosphonate and methylphosphonothioate]. AB - A facile synthesis of adenosine 3', 5'-cyclic methylphosphonate (1) and 3', 5' cyclic methylphosphonothioate (2) was accomplished by treatment of 2'-protected adenosine with O,O-bis (1-benzotriazolyl) methylphosphonate or O,O-bis (1 benzotriazolyl) methylphosphonothioate under mild conditions, respectively. The ability of the compounds to inhibit DNA synthesis in mice hepatoma was tested. A pair of diasteromers of 2'-protected (1) was separated and the configuration at the phosphorus atom was identified. PMID- 2559580 TI - [Effects of imazodan and dazoxiben on cAMP levels and PGI2 production in cultured bovine aortic endothelial cells]. AB - We have investigated the effects of imazodan, a potent inhibitor of phosphodiesterase III (PDE III) and dazoxiben, a selective inhibitor of thromboxane synthetase on cAMP levels and PGI2 production in cultured bovine aortic endothelial cells by radioimmunoassay. When cultured endothelial cells were incubated with imazodan, intracellular levels of cAMP were increased in a dose-dependent manner. PGI2 production induced by arachidonic acid (AA) was not affected by imazodan 0.1-10 mumol/L. But imazodan 100 mumol/L caused a 35% inhibition of PGI2 production. In the presence of AA, dazoxiben could also elevate intracellular levels of cAMP. Furthermore, dazoxiben 1-10 mumol/L caused a marked increase in PGI2 production, but 1000 mumol/L inhibited PGI2 production. PMID- 2559581 TI - [Scavenging effects of probimane on active oxygen free radicals by electron spin resonance]. AB - Probimane, dl-1,2-bis (4-morpholine-methyl-3, 5-dioxopiperazin-1-yl) propane, is a new antitumor agent synthesized by Shanghai Institute of Materia Medica, Chinese Academy of Sciences. The scavenging effects of probimane on active oxygen radicals produced in 3 different systems were studied with the ESR spin trapping methods. In Fenton's reaction, probimane remarkably scavenged hydroxyl radicals (.OH) and the rate of scavenging .OH by probimane 0.05 mmol/L was 47%, compared to 5% by vitamin E (VE) and 30% by ascorbic acid (AA). In irradiation riboflavin system, in which superoxide (O2-.) was produced, the agent also had the scavenging effects on O2(-.). The rate of scavenging O2-. by probimane 0.05 mmol/L was 13%, higher than that by VE (7%) but lower than that by AA (90%). In cell system where the active oxygen radicals were produced during the respiratory burst of human neutrophils (Neu) stimulated by TPA (tetradecanoylphorbol acetate), probimane exhibited a dose-dependent scavenging action on the radicals. The rate of the radical scavenging by probimane 0.05 mmol/L was 37%, much higher than that by VE (9%) but lower than that by AA (68%). Probimane had no effect on the rate of oxygen consumption by human Neu, measured with spin probe oxymetry. PMID- 2559582 TI - DDT toxicity: variations in tissue non-specific phosphomonoesterases and gluconeogenic enzymes in three teleosts. AB - Three fish species were exposed to a sublethal dose (0.35 mg/l) of DDT continuously for a period of 50 days and the effect of hepatic and renal acid and alkaline phosphatases, glucose-6-phosphatase and fructose-1,6-diphosphatase activities was observed at 15, 30 and 45 days. Exposure to DDT at 15 days led to the fall and increase thereafter (at 30 and 45 days) in the activities of acid phosphatase, glucose-6-phosphatase and fructose-1,6-diphosphatase in hepatic tissue, where as alkaline phosphatase in liver registered an increase at 15, 30 and 45 days DDT exposure. In renal tissue the trend of 4 phosphatases was same as that of alkaline phosphatase in the liver. The changes in these 4 phosphatases were more pronounced in C. punctatus than in G. batrachus and L. rohita. PMID- 2559584 TI - alpha-Melanotropin induced excessive grooming involves brain acetylcholine possible interaction in the ventral tegmental area. AB - Alpha-melanotropin (alpha-MSH) injected into the ventral tegmental area (VTA) or intraventricularly (icv) elicits excessive grooming. The icv infusion of the peptide also induces the stretching-yawning syndrome (SYS). The present study demonstrates that intra-peritoneal, icv or VTA administration of atropine, suppresses alpha-MSH-induced behavior elicited by i cv or VTA injections of the peptide. Experimental evidence is presented suggesting that alpha-MSH may act specifically on a cholinergic afferent to the VTA. The results appear to indicate that a neural target distinct from the dopamine system may be formerly activated by the peptide to elicit behavioral changes such as excessive grooming. PMID- 2559583 TI - Renal handling of electrolytes and (Na + K)-ATPase activity after unilateral nephrectomy during long-term ethanol feeding. AB - The effect of long-term ethanol feeding on the activity of (Na + K)-ATPase in cortex and outer medulla and fractional excretion of electrolytes in remnant kidney of adult rats after unilateral nephrectomy were studied. Wistar adult rats were fed 20% (v/v) aqueous ethanol solution as sole drinking fluid for 8-10 weeks. Right kidney was removed under ether anaesthesia. The animals were subjected to an acute NaCl loading by means of a continuous infusion given 2, 7 and 14 days after nephrectomy. Renal handling of electrolytes was estimated from fractional excretion of sodium and potassium. After the infusion the animals were killed and (Na + K)-ATPase and Mg2+-ATPase activities were measured in the cortex and outer medulla of the remnant kidney. Two days after nephrectomy both groups showed a gradual increase of renal (Na + K)-ATPase activity reaching 60 percent at day 14. Mg2+-ATPase activity did not change with respect to basal values. Compared to basal values the fractional excretion of sodium after nephrectomy, dropped in both groups but more significantly in the ethanol-fed rats than in the control group. Fractional excretion of potassium did not change in the control group after nephrectomy while the ethanol-fed group displayed a significative decrease at days 7 and 14. According to our results the rise in renal (Na + K) ATPase activity is consistent with the renal sodium retention found in ethanol fed rats. PMID- 2559585 TI - Adrenal cytochrome-c-oxidase in neonatal and adult rats: sex differences. AB - Cytochrome-c-oxidase was measured in the adrenals of neonatal rats from 1 to 10 days of age and adult rats. No differences between the adrenals of female and male neonatal rats of the same age were found. Adult rats, on the contrary, showed sexual differences. Enzymatic activity was higher in males than in females. In the latter, there were important changes during the estrus cycle. The highest values were observed during diestrus and the lowest during estrus. These results provide new evidence of the close interrelations between gonads and adrenal glands during sexual maturation. PMID- 2559586 TI - Opposite effects of centrally administered neuropeptide Y (NPY) on locomotor activity of spontaneously hypertensive (SH) and normal rats. AB - Centrally administered neuropeptide Y has been shown to produce sedation manifested by a suppression of locomotor activity and a synchronizing effect on the EEG pattern in normal rats. It has been suggested that this sedative effect of NPY is largely due to a facilitation of the alpha 2-adrenergic transmission line. In the spontaneously hypertensive (SH) rat, the NPY-induced up-regulation of alpha 2-adrenoceptors observed in normal rats is absent, and NPY produces a desynchronization of the EEG. In the present study, we have therefore examined the effects of NPY on locomotor activity of SH rats and of inbred controls of the Wistar-Kyoto (WKy) strain, in both morning and evening sessions. In morning sessions, NPY (0.2-5.0 nmol intracerebroventricularly, i.c.v.) increased locomotor activity of SH rats in a dose-related manner. WKy rats were largely inactive per se, and no effects of NPY could be detected. In evening sessions, when spontaneous activity is high, NPY (I.0 nmol i.c.v.) still increased the activity of the SH rats. In WKy rats, an activity suppression similar to that previously reported for normal Sprague-Dawley rats was seen. The present results indicate that the sedative action of NPY in different rats strains correlates with the ability of the peptide to up-regulate alpha 2-adrenergic receptors. PMID- 2559587 TI - Sex-linked differences in cortisol, ACTH and prolactin responses to 5-hydroxy tryptophan in healthy controls and minor and major depressed patients. AB - Some researchers have found that the administration of 5-hydroxytryptophan (5 HTP) results in increased cortisol secretion in major depressives but not in healthy controls. Other authors observed gender-related differences in cortisol responses to 5-HTP in major depressives. In order to investigate the pituitary/adrenal responsivity to 5-HTP, the authors measured cortisol, adrenocorticotropic hormone (ACTH) and prolactin (PRL) in 30 healthy controls and in 90 depressed patients; the hormone levels were determined in baseline conditions and 60, 90 and 120 min after 125 mg L-5-HTP (orally, non-enteric coated). We found that healthy men had significantly higher cortisol responses to L-5-HTP than healthy women. In the major depressives with melancholia and/or psychotic features these differences were reversed: women exhibited significantly higher cortisol and PRL responses than men. In the female group the most severely depressed patients had increased cortisol and PRL responses to L-5-HTP. The amplitudes of the cortisol, ACTH and PRL responses to L-5-HTP were significantly and positively correlated. It was concluded that the central serotonergic regulation of ACTH and PRL is significantly different between the sexes and between healthy controls, minor depressives and severely depressed patients. PMID- 2559588 TI - [Oxygen free radicals in medicine (I)]. AB - Free radicals are highly reactive molecules, and therefore transient, which have an odd number of electrons and are generated in vivo as byproducts of normal metabolism. In this review we survey basic concepts on the chemistry of oxygen free radicals, their cellular sources and the reactions they can undergo. We also discuss the cellular defenses against free radicals induced damage. The disfunction induced by free radicals may thus be a major component of several pathological conditions. The critical role played by free radicals in ageing, carcinogenesis, reperfusion injury and and respiratory distress is reviewed. PMID- 2559589 TI - [Papillary carcinoma of the male breast]. AB - The authors described a breast intraductal papillary carcinoma, of a 73 year old man, with a Paget disease presentation. The pathological features of differential diagnosis were discussed and enunciated the literary review of the etiology and prognosis. PMID- 2559590 TI - [Human chorionic gonadotropin and carcinoembryonic antigen in small cell carcinoma (oat cell) of the urinary bladder]. AB - Patient suffering from vesical carcinoma, which is identified as small cells (oat cells) carcinoma. We carried out a histochemical study with H.C.G. and C.E.A. and analyse the results obtained. PMID- 2559591 TI - Molecular properties of voltage-sensitive calcium channels. PMID- 2559592 TI - Intracellular mechanisms involved in "run-down" of calcium channels. PMID- 2559593 TI - Two types of Ca channels in smooth muscle cells isolated from guinea-pig taenia coli. AB - 1) Two types of voltage dependent Ca channels with different conductances and inactivation kinetics were identified from cell-attached patch clamp recordings. One type, with a larger conductance of 25pS, had a threshold of activation near 40mV and the mean current inactivated slowly. A second type of Ca channel, with a smaller conductance of 12pS channel, but the averaged mean current inactivated rapidly. 2) Cadmium ions inhibited the large conductance Ca channel currents, while the small conductance Ca channel currents was not blocked. 3) Large conductance Ca channel current selectively inhibited by dihydropyridine derivative, Nifedipine. Small conductance Ca channel current is abolished in dose dependent manner by pyrethroid insecticide, tetranethrine. PMID- 2559594 TI - Molecular analysis of calmodulin and smooth muscle myosin light chain kinase. PMID- 2559595 TI - Analysis of calcium-binding sites in calcium-activated neutral protease. PMID- 2559597 TI - Calcium and polyphosphoinositide regulation of actin network structure by gelsolin. PMID- 2559598 TI - Characterization of the calmodulin-binding domain of calcineurin deduced from a complementary DNA clone. PMID- 2559596 TI - In situ phosphorylation of human platelet and rat basophilic leukemia cell (RBL 2H3) myosin heavy chain and light chain. PMID- 2559599 TI - The role of brain protein phosphatases 1 and 2A in the regulation of microtubule assembly. PMID- 2559600 TI - Demonstration of different regional distributions of calcineurin subunits using monoclonal antibodies. AB - Immunohistochemical localizations of calcineurin subunits A (60 KDa) and B (20 KDa) were examined in a rat brain using subunit specific monoclonal antibodies. The immunoreactivity of the subunit A was abundant in the hippocampus, in the striatum, and in the thalamus, but weak in the neocortex. On the contrary, the immunoreactivity of the subunit B was more abundant in the cortex, and it was more ubiquitous than subunit A. The distribution of subunit A in the rat brain very well agreed with that of zinc, which is an intrinsic metal ion and a potent inhibitor of calcineurin phosphatase. PMID- 2559601 TI - Stimulation by inositol trisphosphate and tetrakisphosphate of a protein phosphatase. PMID- 2559602 TI - Regulation of the 63-kDa subunit-containing calmodulin-dependent cyclic nucleotide phosphodiesterase isozyme. PMID- 2559603 TI - Molecular properties of cyclic nucleotide phosphodiesterase isozymes. AB - Mammalian cells contain multiple molecular forms of cyclic nucleotide phosphodiesterase that differ in substrate specificity and kinetic and regulatory properties. Calcium/calmodulin and cyclic GMP are important regulators of the hydrolysis of cyclic AMP by either stimulating or inhibiting the activity of distinct forms of phosphodiesterase. Several isozymes of cyclic nucleotide phosphodiesterase have been purified to apparent homogeneity. Although some sequence homology is observed the isozymes appear genetically distinct by immunological criteria. Cyclic AMP- and calmodulin-dependent protein kinases can phosphorylate these enzymes and alter their kinetic and regulatory properties. Both tissue specificity and pharmacological selectivity of isozymes have been demonstrated for several drugs. In certain cases, e.g. cardiac muscle, the selective inhibition of a high affinity cAMP phosphodiesterase activity in a specific subcellular fraction correlates with pharmacologic responses. The results from molecular and pharmacologic studies of cyclic nucleotide phosphodiesterases have indeed expanded the role this system of isoenzymes exerts in the regulation of cellular function. PMID- 2559605 TI - A possible mechanism for increments of calcium and protein kinase C activity in kidney of experimental acute fluorosis. PMID- 2559604 TI - The calpain-calpastatin system in hematopoietic cells. AB - Calpain I requires low Ca2+ for activation and calpain II requires high Ca2+. It was generally accepted that erythrocytes contain calpain I and calpastatin, but no calpain II. We have recently found, however, that nucleated chicken erythrocytes contain both calpains I and II in addition to calpastatin. The finding is significant in rectifying the previous view that the chicken has only one molecular species of calpain, whereas mammals have two. Another erroneous view which prevailed previously was that polymorphonuclear (PMN) cells contain only one calpain species. We could also recently demonstrate that pig PMN cells do contain both calpains I and II. The cloning of cDNAs for calpastatin enabled us to utilize them as the probes in studying the expression of calpastatin in various hematopoietic cell-line cells. We found that several T cells infected with human retrovirus HTLV-I markedly increased the production of calpastatin, which could be measured both by calpain-inhibition assay and by Western blot analysis, but the level of mRNA for calpastatin did not significantly change when compared with noninfected T cells. The increase in calpastatin protein always parallels with the expression of interleukin 2 receptor protein by the HTLV-I infected T cells, although the biological implication of such phenomena is almost entirely unknown yet. PMID- 2559606 TI - The calcium pump of the plasma membrane: structure-function relationships. PMID- 2559608 TI - Persistence of human papillomaviruses. PMID- 2559607 TI - Molecular structure and function of phospholamban: the regulatory protein of calcium pump in cardiac sarcoplasmic reticulum. PMID- 2559610 TI - Extracellular HSV 2 inhibition of lymphocyte response in healthy subjects. PMID- 2559609 TI - Genetic engineering and properties of novel herpes simplex viruses for use as potential vaccines and as vectors of foreign genes. PMID- 2559612 TI - Possible factors contributing to persistence of cytomegalovirus infection. PMID- 2559611 TI - Expression of herpes simplex virus 1 glycoprotein B in human cells and protection of mice against lethal herpes simplex virus 1 infection. PMID- 2559613 TI - Cell-mediated immunity in infectious mononucleosis. PMID- 2559614 TI - CD4 as the receptor for retroviruses of the HTLV family: immunopathogenetic implications. PMID- 2559615 TI - Strategies for the development of a rotavirus vaccine against infantile diarrhea with an update on clinical trials of rotavirus vaccines. PMID- 2559616 TI - [A clinical study of congenital adrenal hyperplasia]. AB - We have compared three treatments of congenital adrenal hyperplasia (CAH) for their effect on physical development. Thirteen girls and two boys with CAH due to 21-hydroxylase deficiency were treated with three different treatments, hydrocortisone (HC), dexamethasone (DXM) and cyproterone acetate (CA). The results showed that height growth was better with HC and CA than DXM, and bone excessive maturation was more suppressed with DXM and CA than HC. A dose dependent relationship was revealed between body weight and dose of HC. Iatrogenic obesity was found in 42.9% and 38.1% of the patients treated with DXM and HC, but none of the patients treated with CA did became obese. Physical growth was better with CA treatment than HC or DXM treatment, but CA may have a suppressive effect on the pituitary-adrenal axis observed carefully, especially on prepubertal and pubertal cases. PMID- 2559617 TI - [Klippel-Trenaunay-Weber syndrome with papillary cystadenoma of the epididymis: a case report]. AB - A case of Klippel-Trenaunay-Weber syndrome with papillary cystadenoma of the right epididymis is reported. A 7-year-old boy visited our clinic on October 19, 1985 with a tender right intra-scrotal mass (35 X 15 m) and pyuria. In spite of the initial treatment with antibiotics, the tumor grew larger in December, 1987. The patient underwent right orchiectomy on December 23, 1987. Macroscopically, the specimen measured 40 X 23 X 20 mm and weighed 17 g. Histological examinations demonstrated that the tumor was papillary cystadenoma of the right epididymis. The patient lacked the signs of von Hippel-Lindau disease since there were no abnormal findings on brain and abdominal computer tomographic scan, and retinae were normal on fundoscopic examinations. PMID- 2559618 TI - [Extramammary Paget's disease with a large mass in male genitalia: a case report]. AB - A 57-year-old Japanese man presented with a mass 3 cm in diameter at the root of the penis. The patient had noticed the mass growing for 5 years, but had no pain or itching. Histological examination revealed it to be invasive Paget's disease, and the mass was resected along with skin of normal appearance within 3 cm around it. Regional lymph node metastasis was also revealed in the operation, and irradiation at the pelvic and inguinal region was done. Extramammary Paget's disease with a large mass is relatively rare, and invasive extramammary Paget's disease has a poor prognosis. PMID- 2559619 TI - [A case of exohepatic pedunculated hepatocellular carcinoma suspected of adrenal tumor]. AB - A case of exohepatic pedunculated hepatocellular carcinoma that was clinically diagnosed as nonfunctioning adrenal tumor is reported. A 66-year-old man was admitted to our hospital for further examination of unstable angina pectoris. Abdominal echogram and CT scan revealed a large tumor in the right retroperitoneal space. Selective right renal arteriography demonstrated that the tumor was fed by the capsular branch of right renal artery and the right adrenal artery. Adrenal function was normal. Preoperative diagnosis of right nonfunctioning adrenal tumor was made. On operation we found that the tumor was pedunculated from the liver and adhered massively to both right kidney and vena cava. The tumor and right kidney were removed. A histopathological examination demonstrated well differentiated hepatocellular carcinoma (Edmondson's grade II type). PMID- 2559620 TI - [Clinical aspects and diagnosis of an atypical form of genital herpes]. PMID- 2559621 TI - Lentiviral encephalitides in the immature host: a comparison of human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus (SIV) brain infection. AB - The human immunodeficiency virus (HIV) and the simian immunodeficiency virus (SIV) are members of the lentivirus subfamily of retroviruses. Both are capable of establishing persistent infection in the nervous system and both demonstrate increased virulence in the immature of their respective host species. Brain infection with these primate lentiviruses share common virologic and neuropathologic features. The available evidence to date suggests that viral factors may determine neuroinvasiveness and possibly neurovirulence, but that host factors may be largely responsible for the neuropathologic changes observed. PMID- 2559622 TI - Epstein-Barr-virus-related post-bone-marrow-transplant lymphoproliferative disease: association with cytomegalovirus infection and Down syndrome donor marrow. AB - We describe the development of Epstein-Barr-virus (EBV)-related lymphoproliferative disease (LPD) in the recipient of a histocompatible bone marrow transplant (BMT). Although this rare complication is more common in recipients of mismatched bone marrow, several distinguishing features of our case may have contributed to the development of LPD in the recipient of a matched bone marrow transplant. The patient had received marrow from a sibling with Trisomy 21, a syndrome associated with variable cellular and humoral immune defects. Our patient also was infected with cytomegalovirus and was treated with immunosuppressant therapy for graft versus host disease. Although development of LPD in transplant recipients is a multifactorial process, either acquired or congenital immunosuppression/dysregulation is a common prerequisite for the process. Our case suggests that subtle immune defects in individuals with Down syndrome may contribute to the immunosuppressed setting in which EBV-related LPD can develop. PMID- 2559623 TI - Parvovirus B19 infection in patients receiving cancer chemotherapy: the expanding spectrum of disease. AB - Initial reports have suggested that human parvovirus B19 infection in immune compromised individuals may cause prolonged or chronic bone marrow suppression. We report four children receiving cancer chemotherapy who developed atypical patterns of cutaneous and hematologic disease associated with parvoviral infection. B19 infection was demonstrated by specific serologies. This report suggests that patients undergoing myelosuppressive or immunosuppressive therapy may have unusual manifestations of infection and be at risk for complications associated with this common and widespread pathogen. PMID- 2559624 TI - Use of silica gel polymer for DNA extraction with organic solvents. AB - Phenol and chloroform are the standard solvents used for DNA extraction. These solvents aid in the removal of protein and lipid from crude or partially purified cell extracts. Although the procedure is well established, the solvents are noxious, caustic, and unpleasant. We describe in this paper the use of a special blood collection tube to isolate the offensive organic solvents. With the use of silica gel polymer containing tubes, phenol, phenol:chloroform, or chloroform can be separated from the DNA containing aqueous phase in a rapid and safe manner. The method permits higher yields of DNA since the DNA is poured from the tube rather than aspirated with pipet. PMID- 2559625 TI - A sensitive method for detection of calmodulin-dependent protein kinase II activity in sodium dodecyl sulfate-polyacrylamide gel. AB - A procedure for detecting protein kinase activities of the alpha and beta subunits of calmodulin-dependent protein kinase II separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis is described. After electrophoresis, the gel was immersed in 6 M guanidine HCl for 1 h and then in a buffer containing 0.04% Tween 40 for 16 h at 4 degrees C for renaturation of the resolved polypeptides. The renatured polypeptides in the gel were incubated with [gamma 32P]ATP for phosphorylation of either the substrate included in the polyacrylamide gel or the kinase itself. After removal of the unreacted [gamma 32P]ATP, the protein kinase activities were visualized by autoradiography. Two radioactive protein bands of Mr 50,000 and 60,000, which corresponded to the alpha and beta subunits, were detected only when the phosphorylation was carried out in the presence of Ca2+ and calmodulin. Approximately 0.05 micrograms of the enzyme could be detected on a gel containing no protein substrate. When microtubule-associated protein 2 was included in the gel, the sensitivity of the detection of calmodulin-dependent protein kinase II in the gel was more than one order of magnitude higher than that in the gel containing no protein substrate. PMID- 2559626 TI - Enzymatic quantification of sphingosine in the picomole range in cultured cells. AB - An enzymatic method to quantify the mass levels of free sphingosine in cellular lipid extracts was developed. The assay is based upon the observation that ceramide is phosphorylated by Escherichia coli diacylglycerol kinase. Although sphingosine is not recognized by the enzyme, it can be converted to a substrate by acylation with hexanoic anhydride. Using a mixed micellar assay, previously reported for the mass quantification of diacylglycerol, the short-chain ceramide (N-C6-sphingosine), generated by acylation, is quantitatively phosphorylated to N C6-[32P]sphingosine phosphate. This assay allows quantification of sphingosine over a broad range from 25 to 5000 pmol. When this assay was applied to standard compounds, reverse-phase thin-layer chromatography of the reaction products was adequate to separate the phosphorylated derivatives of long-chain ceramide and N C6-sphingosine. However, the presence of other lipids in extracts from biological samples (mainly monoalkylglycerols which are also a substrate for the diacylglycerol kinase) interfered and necessitated an additional purification step. The most efficient purification step devised was a combination of anion- and cation-exchange chromatography. The mass levels of free sphingoid bases in different cultured cells were quantified using this assay. Levels varied between 8 to 20 pmol/10(6) cells. When normalized to phospholipids, sphingosine levels varied between 0.01 and 0.04 mol%. The lowest levels were found in L929 cells, while Schwann cells derived from Twitcher mice contained the highest levels. These levels were significantly higher than those of Schwann cells derived from normal mice. PMID- 2559627 TI - [3H]PN200-110 and [3H]ryanodine binding and reconstitution of ion channel activity with skeletal muscle membranes. AB - Skeletal muscle membranes derived either from the tubular (T) network or from the sarcoplasmic reticulum (SR) were characterized with respect to the binding of the dihydropyridine, [3H]PN200-110, and the alkaloid, [3H]ryanodine; polypeptide composition; and ion channel activity. Conditions for optimizing the binding of these radioligands are discussed. A bilayer pulsing technique is described and is used to examine the channels present in these membranes. Fusion of T-tubule membranes into bilayers revealed the presence of chloride channels and dihydropyridine-sensitive calcium channels with three distinct conductances. The dihydropyridine-sensitive channels were further characterized with respect to their voltage dependence. Pulsing experiments indicated that two different populations of dihydropyridine-sensitive channels existed. Fusion of heavy SR vesicles revealed three different ion channels; the putative calcium release channel, a potassium channel, and a chloride channel. Thus, this fractionation procedure provides T-tubules and SR membranes which, with radioligand binding and single channel recording techniques, provide a useful tool to study the characteristics of skeletal muscle ion channels and their possible role in excitation-contraction coupling. PMID- 2559628 TI - A fluorescent oligopeptide energy transfer assay with broad applications for neutral proteases. AB - A fluorescent peptide substrate to explore the protease specificity for the amino acid regions C- and N-terminal to the cleavage site has been designed. Intramolecular quenching of indole fluorescence by an N-terminal dansyl group separated by six amino acid residues forms the basis of this assay. For a particular enzyme, specificity can be designed into the peptide sequence by means of the number of residues that separate the two chromophores. In the present instance, the heptapeptide Dns-Gly-Lys-Tyr-Ala-Pro-Trp-Val is used to assay angiotensin converting enzyme (ACE), Astacus protease, carboxypeptidase A, alpha chymotrypsin, and trypsin, all of which cleave the peptide in accord with their known specificity: Trypsin and Astacus protease hydrolyze only the Lys-Tyr and Tyr-Ala bonds, respectively. alpha-Chymotrypsin primarily cleaves the Tyr-Ala bond while ACE makes three successive dipeptidyl cleavages from the C-terminus. Carboxypeptidase rapidly hydrolyzes first the Trp-Val and then the Pro-Trp bond. For all of the enzymes, catalytic activity (kcat/Km) is in the range from 10(5) to 10(6) M-1 s-1. Hydrolysis causes a fluorescence increase in the 310 to 410 nm region of 8.6- to 13.6-fold depending on the enzyme that is assayed. Assays can be designed based on the increase in tryptophan fluorescence or by individual product analyses using thin-layer or high-performance liquid chromatography. The specificity and sensitivity of such internally quenched fluorescent oligopeptides would seem to be ideal for the assay of specific endoproteases. PMID- 2559629 TI - Mapping introns by exon excision. AB - A direct method for mapping introns has been devised. The technique makes use of a radioactive synthetic RNA transcript of the gene and a complementary, single stranded DNA copy of mRNA-derived sequences. Upon hybridization of the cDNA to RNA and cleavage with ribonuclease H, only exonic RNA sequences are degraded. The surviving RNA fragments are the introns. Electrophoretic analysis in denaturing agarose gels reveals the number and size of the introns. The order of the introns is determined separately using unlabeled RNA transcripts; surviving RNA fragments are transferred to a solid support and the blot is probed sequentially with a nested set of genomic RNA transcripts of the opposite strand. Using the human prothymosin alpha gene as an example, four introns were identified which from 5' to 3' were 2.6, 0.47, 0.47, and 0.28 kb in size. From mapping and sequencing experiments the sizes are 2.6, 0.465, 0.459, and 0.295 kb, respectively. Similarly, the presence of two 300-bp insertions in a human prothymosin alpha pseudogene was established; the inserts were later identified as 295-bp Alu repetitive elements. PMID- 2559631 TI - Effects of single intravenously administered doses of dexamethasone on response to the adrenocorticotropic hormone stimulation test in dogs. AB - The effects of single IV administered doses of dexamethasone on response to the adrenocorticotropic hormone (ACTH) stimulation test (baseline plasma ACTH, pre ACTH cortisol, and post-ACTH cortisol concentrations) performed 1, 2, and 3 days (experiment 1) or 3, 7, 10, and 14 days (experiment 2) after dexamethasone treatment were evaluated in healthy Beagles. In experiment 1, ACTH stimulation tests were carried out after administration of 0, 0.01, 0.1, 1, and 5 mg of dexamethasone/kg of body weight. Dosages greater than or equal to 0.1 mg of dexamethasone/kg decreased pre-ACTH plasma cortisol concentration on subsequent days, whereas dosages greater than or equal to 1 mg/kg also decreased plasma ACTH concentration. Treatment with 1 or 5 mg of dexamethasone/kg suppressed (P less than 0.05) post-ACTH plasma cortisol concentration (on day 3 after 1 mg of dexamethasone/kg; on days 1, 2, and 3 after 5 mg of dexamethasone/kg). In experiment 2, IV administration of 1 mg of dexamethasone/kg was associated only with low (P less than 0.05) post-ACTH plasma cortisol concentration in dogs on day 3. In experiment 2, pre-ACTH plasma cortisol and ACTH concentrations in dogs on days 3, 7, 10, and 14 and post-ACTH plasma cortisol concentration on days 7, 10, and 14 were not affected by dexamethasone administration. The results suggest that, in dogs, a single IV administered dosage of greater than or equal to 0.1 mg of dexamethasone/kg can alter the results of the ACTH stimulation test for at least 3 days. The suppressive effect of dexamethasone is dose dependent and is not apparent 7 days after treatment with 1 mg of dexamethasone/kg.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559630 TI - Evaluation of antithrombin-III activity as a coindicator of disseminated intravascular coagulation in cats with induced feline infectious peritonitis virus infection. AB - Six adult specific-pathogen-free cats were inoculated intraperitoneally with a cell culture-adapted strain of feline infectious peritonitis virus. Plasma samples were evaluated for antithrombin-III (AT-III) activities at post inoculation days (PID) 0, 4, and 11 and at termination on PID 16 (1 cat) or 21 (5 cats). Other hemostatic values evaluated were activated partial thromboplastin times, prothrombin times, thrombin times, fibrinogen, platelet counts, and fibrin/fibrinogen degradation products. Antithrombin-III activity remained within normal or above normal range (89 to 246%) in all cats, with the exception of one cat on PID 4 (AT-III, 70%). Mean baseline AT-III activity for 6 cats at PID 0 was 123%. Mean AT-III activity on PID 4, 11, and 16 or 21 was 98, 162, and 130%, respectively. On PID 4 and 16 or 21, results of coagulation screening tests indicated that all cats had disseminated intravascular coagulation. Histologically, cats also had severe fibrinonecrotizing thrombovasculitis. PMID- 2559632 TI - Prevalence of bovid herpesvirus-4 and its antibody in cattle in Minnesota. AB - Serologic analyses and virus isolation studies were carried out to determine the role of bovid herpesvirus-4 (BHV-4) in infections in cattle, principally those of the reproductive tract. Serologic analyses were performed, using an indirect fluorescent antibody test on thoracic fluid specimens from aborted fetuses and on sera from 3 sources of adult cattle. Virus isolation was attempted from field cases of abortion, early embryo death, and postpartum vulvovaginitis/metritis, using uterine discharge and buffy coat preparations obtained from cows and tissues obtained from aborted fetuses. Of 420 fetal thoracic fluid specimens examined, 5 were positive for BHV-4 antibodies. Seventeen percent of adult cattle from 2 sources ie, clinically normal herds and abattoir cattle, were seropositive for BHV-4 antibodies. Cattle from a third source, 4 herds with high incidence of reproductive tract disorders, had a seroprevalence rate between 36 and 88%. Two isolates of BHV-4 were also obtained from this group. The overall incidence of BHV-4 antibodies in clinically normal cattle was higher than previously recognized, with relatively higher prevalence in herds having reproductive problems (chi 2 = 156.5, P less than 0.005). At least 10% of the BHV-4 antibody positive sera did not have neutralizing antibody against bovine viral diarrhea virus and/or bovid herpesvirus-1, both important causes of bovine reproductive tract disorders. PMID- 2559633 TI - Immunohistologic study of pulmonary and lymphatic tissues from gnotobiotic pigs inoculated with ara-T-resistant strain of pseudorabies virus. AB - Ara-T-resistant strain of pseudorabies virus (PRV) was inoculated intranasally into six 2-week-old gnotobiotic pigs. Five inoculated pigs were sneezing and coughing. In pigs 1 to 4 killed on postinoculation days (PID) 3, 5, 7, and 9, respectively, PRV antigen was detected in respiratory epithelial cells, and pigs had severe pneumonitis. In pigs 5 and 6 killed on PID 11 and 13, respectively, PRV antigen was localized in macrophages in alveoli and necrotizing nodules. Immunoglobulin-containing cells (IgG, IgM, and IgA) were detected first in pneumonic lesions in pig 4 killed on PID 9. Detection of immunoglobulin containing cells was coincident with pulmonary inflammation and regeneration of pneumonic lesions. The number of IgG-containing cells was greater than that of IgM- and IgA-containing cells. Corresponding to transient viral multiplication, IgG-, IgM-, and IgA-containing cells were demonstrated first in lymphatic tissues in pig 1 killed on PID 3 and their number was 5 to 10 times more than those in control pigs 7 and 8. Seemingly, PRV replication in lymphatic tissues stimulated the proliferative response of specific immunoglobulin-producing cells, and the appearance of immunoglobulin-containing cells in the lungs was associated with clearance of PRV and regeneration of pneumonic lesions. PMID- 2559635 TI - Experimental rotavirus infection in three-week-old pigs. AB - Thirteen 3-week-old pigs that had been allowed to nurse for the first 16 to 18 hours after birth were orally inoculated with 1 x 10(6.5) TCID50 of porcine rotavirus. All developed diarrhea, anorexia, and vomiting by postinoculation (PI) hour 30. These signs had abated by PI day 6. Villus blunting in the small intestine was most severe in the jejunum and ileum of pigs euthanatized between PI days 3 and 5. Villi had returned to nearly normal length by PI day 6, although fused villi were seen in a few locations in the distal portion of the jejunum and in the ileum. Virus was detected in the feces of inoculated pigs by isolation in cell cultures and by electron microscopy during the 7-day course of the experiment. There was 1 extraintestinal virus isolation from the lung of 1 pig at PI day 2. Infection and disease developed in the presence of serum-neutralizing antibody obtained by nursing seropositive sows. There was no significant change in neutralizing antibody titers in the 3-week-old pigs over the course of the experiment. In this experimental work, a model to study rotavirus infection in 3 week-old pigs has been developed. PMID- 2559634 TI - Further phenotypic characterization of target cells for bovine leukemia virus experimental infection in sheep. AB - To determine the phenotype of target cells for bovine leukemia virus (BLV) infection in sheep, we analyzed blood lymphocytes from BLV-infected clinically healthy and leukemic sheep by use of monoclonal antibodies. In clinically healthy and leukemic sheep that were BLV-infected, the blood concentration of T lymphocytes was within normal values, but the number of B lymphocytes was increased in several cases. In addition, the number of blood lymphocytes expressing the BLV antigen correlated well with that of B lymphocytes. Double immunofluorescence staining demonstrated that lymphocytes expressing BLV antigens bore B-cell but not T-cell surface markers. Moreover, neoplastic cells in the lymph nodes of leukemic sheep were stained immunohistochemically with an anti-B monoclonal antibody but not with any of anti-T monoclonal antibody tested, indicating that tumor cells are of B-lymphocyte origin. Collectively, these results show that BLV antigen-positive cells obtained from BLV-infected sheep that have no clinical signs and BLV-induced lymphosarcoma cells belong to the B lymphocyte lineage. PMID- 2559636 TI - Rotavirus replication in colostrum-fed and colostrum-deprived pigs. AB - A porcine rotavirus isolate was titrated in neonatal colostrum-fed and colostrum deprived pigs. The stock rotavirus suspension had a titer of 10(-6.5)/ml and was in its fifteenth cell culture passage in MA-104 cells. Fourteen colostrum-fed pigs were orally inoculated with dilutions of the stock virus suspension ranging from undiluted to 10(-5). These pigs did not develop notable clinical signs during the 7-day experimental trial and no pathologic changes were found in intestine, liver, lung, kidney, spleen, or brain. However, rotavirus was detected in feces of the colostrum-fed pigs, using virus isolation and electron microscopic techniques. Rotavirus was also isolated from lung, brain, or spleen of 4 of 12 of these pigs. Sixteen colostrum-deprived pigs were orally inoculated with dilutions of the stock virus suspension ranging from 10(-1) to 10(-8). Diarrhea developed in 10 of 12 pigs that were given up to the 10(-6) dilution. Seven of these 12 pigs died because of the severity of diarrhea. Pigs that died of rotavirus-induced diarrhea had severe villus loss in the jejunum and ileum. Villi of the small intestine of colostrum-deprived pigs that survived the severe diarrhea were within normal limits at the end of the 7-day trial. The colostrum deprived pigs that were inoculated with a dilution less than 10(-6) and survived past 96 hours underwent seroconversion. Rotavirus was detected by virus isolation and electron microscopy in the feces of all colostrum-deprived pigs that survived beyond 18.5 hours after inoculation. Virus was isolated from lungs, brain, or spleen of 12 of 16 colostrum-deprived pigs. PMID- 2559637 TI - [Respiratory chain diseases in infancy. Clinical presentation and diagnosis]. AB - Nineteen children with defects of the mitochondrial respiratory chain are described. First symptoms appeared during the first two years of their lives. Four types of clinical pictures were identified: 1/neonatal hypotonia. 2/cardiomyopathy. 3/progressive neurological deterioration. 4/multisystem disease. A study of pyruvate and fatty acids metabolism and a skeletal muscle biopsy were performed in all cases. Elevation of beta hydroxybutyrate/acetoacetate and lactate were the most frequent biochemical abnormalities. Muscular biopsy in light microscopy showed in most cases abnormal lipid storage. PMID- 2559639 TI - Concurrent adenoviral and herpetic ocular infections. AB - We report two cases of a previously unrecognized clinical entity: concurrent adenoviral and herpetic ocular infection. The first case was recognized clinically by the presence of herpetic keratitis and findings attributable to epidemic keratoconjunctivitis (EKC) including lid swelling, pseudomembranous and follicular conjunctivitis, subconjunctival hemorrhages, preauricular lymphadenopathy, and corneal infiltrates. Only Herpes simplex virus was recovered by culture. In the second case of apparent EKC (again initially only Herpes simplex was isolated) after maintaining virus cultures long enough to isolate adenovirus, the diagnosis of concurrent infection was established when the second virus was isolated. PMID- 2559638 TI - [Papular acrodermatitis associated with initial Epstein-Barr virus infection]. PMID- 2559640 TI - [Fabry's disease and Klippel-Trenaunay syndrome of the 4 limbs]. PMID- 2559641 TI - [Ultrastructural histology and virology in a case of epithelial focal hyperplasia or Heck's disease]. PMID- 2559643 TI - [Eccrine porocarcinoma]. PMID- 2559642 TI - [Cutaneous eccrine mucinous adenocarcinoma]. PMID- 2559644 TI - [Angiomatoid histiocytofibroma]. PMID- 2559645 TI - [Cutaneous manifestations of POEMS syndrome]. PMID- 2559646 TI - [Rotaviruses: structure and function of the principal polypeptides]. AB - Polypeptides have been defined by studying structural and nonstructural proteins. The rotavirus outer capsid is made up of three proteins: VP7, VP3 and VP9. VP7 is a glycoprotein involved in cell attachment and viral maturation. VP3 is associated with hemagglutination and trypsin activation of virus infectivity; both contain type-specific neutralization determinants. A biological function has not yet been completely defined for VP9. VP6, the main protein of the inner capsid is necessary for mRNA synthesis by the viral transcriptase and determines the subgroup antigenic specificity. These two capsids surround the core which consists of three proteins VP1, VP2, and the product of segment 3, associated with RNA polymerase. Four non-structural polypeptides have been identified (NCVP5, NCVP4, NCVP2, NCVP3); very little is known about their biological role. PMID- 2559647 TI - [Treatment of small hepatocarcinoma, complicating cirrhosis, by total resection of the liver and transplantation]. PMID- 2559649 TI - Advantages of IRMA over RIA in the measurement of ACTH. AB - A technically simple and rapid two-site immunoradiometric assay (IRMA) for human ACTH, based on monoclonal antibodies (MAbs), was compared with a clinically validated ACTH radioimmunoassay (RIA). Both methods measure ACTH 1-39 in unextracted plasma and cross-react less than 0.5% with ACTH fragments. ACTH levels were assessed in 103 patient samples: for concentrations in the range 5.3 1000 ng/L, results by the two methods were significantly correlated (r = 0.82, n = 86, P less than 0.001). The IRMA was more sensitive and had a wider working range than the RIA (detection limits 5.3 ng/L (IRMA) vs 11 ng/L (RIA); CV less than 10% between 19 and 1000 ng/L (IRMA) and CV less than 15% between 30 and 400 ng/L (RIA). In two patients for whom discrepant results were obtained, measurement of ACTH by bioassay and ACTH precursors by direct IRMA demonstrated the greater accuracy of the ACTH IRMA result. The improved performance of the IRMA combined with its many practical advantages compared to RIA, make it ideal for use in detailed clinical and physiological studies which have previously been hampered by the poor reliability of ACTH measurement. PMID- 2559648 TI - [Muco-epidermoid cancer of the esophagus. Review of the literature. Report of a case with 9 years' follow-up]. AB - Mucoepidermoid carcinoma of esophagus is a distinctly uncommon neoplasm. Diagnosis depends on precise microscopic criteria. Microscopic features should show distinct epidermoid features with evidence of intracellular mucus secretion. Their origin is generally believed to be esophageal submucosal glands or their ducts. In a survey of the literature, only 15 cases were found to have been reported. A case of a 43 year old man with a 9 year follow-up is presented. Prognosis of mucoepidermoid carcinoma is the same as ordinary squamous cell carcinoma of the esophagus. PMID- 2559650 TI - Methods of proximal amalgam overhang removal--a comparison of different techniques. AB - It is widely accepted that the overhanging restoration is an aggravating factor in the initiation and progression of periodontal disease, and when detected, the overhang should be removed. Some methods for removing proximal amalgam overhangs are the EVA reciprocating motor driven system, roto-pro, flame-shaped steel bur, diamond bur and sonic scaler. This article illustrates the trimmed surface after overhang removal with photographs taken at 1:1 magnification. The smoothest surface was produced by EVA system where the overhang was removed with a diamond tip followed by polishing with prophylaxis paste using the plastic tip. The roughest surface was produced by the sonic scaler, whilst the others were intermediate. Of the single step methods, the instrument which combined speed while leaving a reasonable surface was the rotopro. PMID- 2559651 TI - Plasma glucose and insulin, urinary catecholamine and cortisol responses to test breakfasts with high or low fibre content: the importance of the previous diet. AB - A 60-hour high-carbohydrate (high-CHO) diet or a 36-hour low-carbohydrate (low CHO) diet was followed by 24 healthy women in a cross-over design to modify liver glycogen content. Thereafter each subject was given a high-sucrose breakfast, a high-protein, high-fibre breakfast or no breakfast. The two different breakfasts evoked larger plasma glucose responses following the low-CHO diet than when following the high-CHO diet. When the two breakfasts followed the same pre-period diet, no significant differences were observed. We conclude that the composition of the previous diet influences the postprandial response to meals and that a standardised diet shortly before test meal studies is of importance for the results in this type of studies. PMID- 2559652 TI - The population genetics of Drosophila transposable elements. PMID- 2559653 TI - Maize transposable elements. PMID- 2559655 TI - In vitro and in vivo activities of WIN 54954, a new broad-spectrum antipicornavirus drug. AB - WIN 54954 (5-[5-[2,6-dichloro-4-(4,5-dihydro-2-oxazolyl)phenoxy]pentyl]-3- methylisoxazole) is a new member of the class of broad-spectrum antipicornavirus compounds known to bind in a hydrophobic pocket within virion capsid protein VP1. In plaque reduction assays, WIN 54954 reduced plaque formation of 50 of 52 rhinovirus serotypes (MICs ranged from 0.007 to 2.2 micrograms/ml). A concentration of 0.28 microgram/ml was effective in inhibiting 80% of the 52 serotypes tested (EC80). WIN 54954 was also effective in inhibiting 15 commonly isolated enteroviruses, with an EC80 of 0.06 microgram/ml. Furthermore, WIN 54954 was effective in reducing the yield of two selected enteroviruses in cell culture by 90% at concentrations approximately equal to their MICs. The therapeutic efficacy of intragastrically administered WIN 54954 was assessed in suckling mice infected with coxsackievirus A-9 or echovirus type 9 (Barty) 2.5 days prior to initiation of therapy. Single daily doses of 2 and 100 mg/kg protected 50% of the mice from developing paralysis (PD50) following infection with coxsackievirus A-9 and echovirus-9, respectively. At the PD50 doses for these two viruses, levels of WIN 54954 in serum were maintained above the in vitro MICs for a significant portion of the dosing interval. The dose-dependent reduction in viral titers observed in coxsackievirus A-9-infected mice correlated well with the therapeutic dose response. The potency and spectrum of WIN 54954 make it a potentially useful compound for the treatment of human enterovirus and rhinovirus infections. PMID- 2559656 TI - Cytomegalovirus infection in immunocompromised guinea pigs: a model for testing antiviral agents in vivo. AB - An experimental model for testing antiviral agents against severe cytomegalovirus (CMV) infection in immunocompromised hosts was developed. The model consisted of cyclophosphamide (Cy) treatment of CMV-infected guinea pigs to simulate CMV infection in immunodeficient individuals. Of the 3 Cy regimens tested, a single 300 mg/kg dose administered one day after virus inoculation resulted in the most severe CMV infection considering mortality rates, mean day of death and loss of body weight. Evaluation of responses to both T and B cell mitogens suggested that the severe and lethal CMV infection resulted from the combined immunosuppressive effect of Cy and CMV. The nucleoside analog [9-(1-3-dihydroxy-2 propoxymethyl)guanine (DHPG) was used to assess the usefulness of the CMV infected immunocompromised host model. DHPG (100 mg/kg/day for 8 days) prevented death but did not reduce virus infectivity titers in blood of Cy-treated, CMV infected guinea pigs. This model of CMV infection in immunocompromised guinea pig is a relevant and convenient experimental tool for the assessment of candidate anti-CMV agents under well-defined experimental conditions, such as appropriate CMV inoculum and Cy regimen. PMID- 2559657 TI - Increased efficacy of ganciclovir in combination with foscarnet against cytomegalovirus and herpes simplex virus type 2 in vitro and in vivo. AB - In tissue culture, efficacy against either murine CMV or HSV-2 was increased 27 fold for the acyclic nucleoside ganciclovir and 3-fold for foscarnet (trisodium phosphonoformate) when the 2 drugs were combined; whereas against human CMV, efficacy was increased 3-fold for both drugs. In mice, efficacy was increased 2 fold for ganciclovir and 4- to 5-fold for foscarnet when used in combination against either murine CMV or HSV-2. These results suggest an additive interaction between the two drugs in vivo. PMID- 2559659 TI - Electron paramagnetic resonance imaging. PMID- 2559654 TI - Biochemical characteristics and physiological significance of major DNA topoisomerases. PMID- 2559658 TI - In vivo protocol for testing efficacy of hand-washing agents against viruses and bacteria: experiments with rotavirus and Escherichia coli. AB - Ten antiseptic formulations, an unmedicated liquid soap, and tap water alone were compared for their capacities to eliminate human rotavirus from the finger pads of adult volunteers; three of the antiseptics, the soap, and the tap water alone were also tested against Escherichia coli. A fecal suspension of virus or bacterium was placed on each finger pad and air dried. The contaminated site was exposed to the test product for 10 s, rinsed in tap water, and dried on a paper towel. The residual virus or bacterium was then eluted. Selected agents were also tested by an analogous whole-hand method by which the entire palm surfaces of both hands were contaminated. Alcohols (70%) alone or with Savlon reduced the virus titer by greater than 99%, whereas the reductions by Proviodine, Dettol, and Hibisol ranged from 95 to 97%. Aqueous solutions of chlorhexidine gluconate were significantly less effective for virus removal or inactivation than 70% alcohol solutions. Furthermore, Savlon in water (1:200) was found to be much less effective in eliminating the virus (80.6%) than the bacterium (98.9%). The tap water alone and the soap reduced the virus titers by 83.6 and 72.5% and the bacterial titers by 90 and 68.7%, respectively. The results of the whole-hand method agreed well with those of the finger pad protocol. We conclude that the finger pad method is a suitable model for testing the in vivo efficacy of hand washing agents and emphasize the need for using appropriate test viruses and bacteria. PMID- 2559660 TI - Nuclear relaxation and Overhauser effect studies of enzyme-substrate interactions. PMID- 2559661 TI - Comparative effects of inhaled silica or synthetic graphite dusts on rat alveolar cells. AB - Alterations in population profiles, morphology, and phagocytic activity of lung cells following in vivo exposure of Fischer 344 rats to a single dose of silica, were compared to similar changes following exposure to synthetic graphite. The effect of silica exposure on alveolar macrophage mycocidal activity was also measured. Acute exposure by inhalation to 100 mg/m3 silica dust produced persistent pulmonary inflammation, as well as long-term changes in pulmonary alveolar macrophages (AM) typical of macrophage activation. Both particles were steadily cleared from the lungs; AM from the silica study, as well as from the 100 and 500 mg/m3 graphite studies, contained ingested particles throughout the entire 3 month experimental period. Lavaged pulmonary cells showed no reduction in viability as a result of exposure to either of the dusts. After silica inhalation, the following changes were observed constantly in bronchopulmonary lavage cell samples: increased numbers of polymorphonuclear leukocytes (PMN), hypersegmented PMN, binucleated AM, increased AM phagocytic indices and capacities, and increased mycocidal activity. Following inhalation of 1-100 mg/m3 synthetic graphite the above-mentioned effects were not seen. Exposure to 500 mg/m3 graphite produced transient inflammation and AM activation for about 24-48 hr. PMID- 2559662 TI - The rate of mitosis in cystosarcoma phyllodes (phyllodes tumor, WHO) of the breast. An analysis of 47 cases. AB - The rate of mitosis was determined in 47 cases of cystosarcoma phyllodes which had already been classified by morphological criteria (tumor margin and cellular atypia) as benign (17 cases), malignant (23 cases) and borderline (7 cases). Tumors which had been classified histopathologically as benign or of borderline had mitotic rates of 3 mitosis/10 HPF or less, whereas malignant cystosarcomas showed rates of 2-36 mitoses/10 HPF. Only high rates of mitosis (10/10 HPF) correlated well with a histopathologic diagnosis of malignancy. PMID- 2559664 TI - Treatment of pigmented villonodular synovitis of the knee with yttrium-90 silicate: prospective evaluations by arthroscopy, histology, and 99mTc pertechnetate uptake measurements. AB - The diffuse form of pigmented villonodular synovitis of eight knee joints of eight patients was treated by intra-articular injection of 185 MBq yttrium-90 silicate (90Y). Six patients had a recurrence of disease after one or two surgical synovectomies. After treatment with 90Y once or twice four knees showed clinical improvement with an accompanying decrease of the inflammatory activity as measured by the technetium-99m pertechnetate (99mTcO4-) uptake ratio and the severity of the diseased synovial tissue. Arthroscopy was performed before and six months after each 90Y treatment. The ratio of 99mTcO4- uptake in the inflamed compared with the normal knee joint correlated well with the macroscopical grading of pigmented villonodular synovitis. In all cases areas of persistent synovitis were found after the 90Y injection and this was confirmed both by histological examination and 99mTcO4- uptake measurements. Biopsy specimens taken from the diseased synovial areas showed histologically mostly less prominent and less numerous villi. The cartilage damage was slightly increased in only two cases. No radiological deterioration was found during follow up (mean 24 months, range 12-41). No complications of the radiosynoviortheses were noted. PMID- 2559665 TI - Reflex sympathetic dystrophy associated with large cell lung carcinoma. PMID- 2559663 TI - [Signet ring cell adenocarcinoma of the bladder]. AB - A case of primary signet-ring cell adenocarcinoma of the urinary bladder is described. The etiopathogenesis, pathological and clinical features, prognosis and treatment of this tumor type are discussed and the literature reviewed. PMID- 2559666 TI - The ordinary light microscope: an appropriate tool for provisional detection and identification of crystals in synovial fluid. AB - To determine if the ordinary light microscope is a useful tool in the detection and identification of crystals in synovial fluid 63 fluids (13 with monosodium urate (MSU), 14 with calcium pyrophosphate dihydrate (CPPD), one with both types of crystal, and 35 without crystals) were examined blindly by two observers, first with an ordinary light microscope, then with a polarising light microscope. Tentative identification of the crystals by their shape was attempted. The sensitivity of the ordinary light microscope for the detection of crystals was 96.2% and 100%, with specificity of 100% and 97.1% for observers 1 and 2, both with respect to the polarising light microscope. The concordance between observers for the ordinary light microscope was chi = 0.90 (0.897, 0.902) and for the polarising light microscope chi = 0.96 (0.958, 0.961). The specificity for identification of MSU and CPPD crystals by their shape with respect to the polarising light microscope was 92.3% and 85.7% for observers 1 and 2. When a polarising light microscope is not available examination of synovial fluid by an ordinary light microscope may allow a provisional diagnosis; definitive identification of the crystals requires a polarising light microscope. PMID- 2559667 TI - [An immunoenzyme method of isolation of foot-and-mouth disease virus by using beta-lactamase conjugate with virus-specific antibodies]. AB - It was shown that in was feasible to use conjugates of virus-specific antibodies and beta-lactamase from Bacillus licheniformis 749/c to identify aphthosa virus antigens. The antigen titers determined by enzyme immunoassay (EIA) using a beta lactamase conjugate were 5-64 times higher than the analogous indices of the complement fixation test. Unlike EIA, that by using the antibody conjugates with peroxidase or alkaline phosphatase there were observed no "background" responses. PMID- 2559668 TI - Cardioprotective effect of trimetazidine and nifedipine in guinea-pig hearts subjected to ischaemia. AB - The cardioprotective effects of nifedipine and trimetazidine were evaluated in the Langendorff heart, the working heart and the heart-lung preparation of the guinea-pig. The effects of pretreatment with the drugs on cardiac function and on high energy phosphate content after global ischaemia were determined. Nifedipine was 1000-fold more active than trimetazidine in depressing cardiac functional parameters. In the Langendorff heart, nifedipine, but not trimetazidine, increased the coronary flow. With the exception of trimetazidine (4 x 10(-5) mol/l) in the Langendorff heart, nifedipine and trimetazidine markedly improved and in a similar fashion, the recovery of cardiac mechanical parameters in the three models of myocardial ischaemia. Pretreatment with nifedipine, but not with trimetazidine, resulted in slightly but statistically not significant higher ATP levels after global ischaemia in the working heart and in the heart-lung preparation. After reperfusion, the cardioprotective effect of nifedipine was associated with increased ATP-levels, whereas that of trimetazidine was merely related to increased phosphocreatine (PCr) levels. In the pithed rat, trimetazidine did not influence the vasoconstrictor response to the selective alpha 1-adrenoceptor agonist cirazoline or to the selective alpha 2-adrenoceptor agonist B-HT 920. The latter response is known to be strongly dependent on the influx of extracellular calcium. Nifedipine caused a pronounced inhibition of vasoconstriction evoked by B-HT 920. These results indicate that trimetazidine is devoid of calcium entry blocking activity. The data obtained clearly show that trimetazidine exerts a pronounced beneficial effect on the ischaemic myocardium without influencing hemodynamic parameters. The mechanism of the cardioprotective activity of the drug is not related to calcium entry blockade or alpha adrenoceptor antagonism, but rather caused by a mechanism at a cellular level, so far not known in detail. PMID- 2559670 TI - [Physiopathology of the migraine crisis and its treatment with a 5-HT1-like agonist]. PMID- 2559669 TI - Evidence for a direct noncholinergic effect of an organophosphorous compound on guinea-pig papillary muscles: are ventricular arrhythmias related to a Na+/K+ ATPase inhibition? AB - The purpose of this study was to determine whether an anticholinesterase organophosphorous compound, methylphosphonothiolate (VX), could display direct effects on isolated guinea-pig ventricular muscle, and to compare these possible effects with those of carbachol (CCH) and physostigmine (PHYS). Our results confirm the direct positive inotropic effect of CCH (stimulation frequency; 2 Hz) in a concentration range from 10(-7) to 10(-3) M; lack of PHYS or VX-induced modifications was set. In the presence of an adrenergic agonist, isoproterenol (ISO) 10(-7) M, CCH or PHYS modulated the positive inotropic effect of ISO. Even with elevated concentrations of CCH (10(-4) M) or PHYS (10(-3) M), arrhythmias were never depicted. VX-induced modifications are different. Under VX, the development of (1) a positive inotropic effect and (2) two contractile events in response to each stimulation were observed. Electrophysiological studies revealed that VX led to the development of delayed after-depolarizations, and eventually triggered activity. We conclude that, in addition to its anticholinesterase activity, VX could induce a Na+/K+ ATPase inhibition. This effect could be at the onset of ventricular arrhythmias that are observed in vivo in organophosphorous compounds poisoning. PMID- 2559671 TI - In vitro studies into some parameters of protein and carbohydrate metabolism in lymphocytes infected with bovine leucosis virus. AB - Several parameters of protein and carbohydrate metabolism were determined in normal and leukemic lymphocytes in vitro in cattle, including arylamidase activity toward beta-naphthylamides of L-amino acids. The homogenate of bovine leukemic lymphocytes, in comparison with the control revealed increase of gamma glutamyltransferase, activity trypsin inhibitor and papain inhibitor concentration and aldolase activity. On the other hand, proteolytic activity toward casein and histomucoid content decreased. Out of the 7 substrates used in the study, only 2, alanyl-beta-naphthylamide and leucyl-beta-naphthylamide, demonstrated lower activity in the leukemic material. Disorders in carbohydrate and protein metabolism in the observed lymphocytes in vitro in cattle are presented in the paper. PMID- 2559672 TI - Detection of antibodies to bovid herpesvirus 2 infection of cattle in Syria. AB - Neutralising antibody to bovid herpesvirus 2 was demonstrated in the serum of 31 (10.8%) of 286 heads of cattle in north, south, and west Syria. 38% of titres were 1:2 to 1:8. There is no published report on isolation of this virus in Syria. PMID- 2559674 TI - Effect of disodium cromoglycate on deuterium oxide-induced degranulation of human eosinophils in allergic asthmatic patients in vitro. AB - Eosinophils from many atopic asthmatic patients have been shown to degranulate in response to 44% deuterium oxide, in contrast to eosinophils from healthy persons. Several pieces of evidence have been obtained recently, indicating that disodium cromoglycate (DSCG) may act on eosinophils. In the present study we show, that prior incubation of eosinophils obtained from the blood of 26 of 30 atopic asthmatic deuterium oxide-responders with DSCG significantly inhibited D2O induced eosinophil degranulation in the dose dependent manner. Eosinophil degranulation was assayed using haemacytomether method. These results suggest, that DSCG may have direct action on eosinophils. PMID- 2559673 TI - [Immunization of young cattle with gp51 of the bovine leukosis virus and the subsequent experimental infection]. AB - Calves were immunised by means of a native Al(OH)3-absorbed gp51-preparation which had been obtained from a foetal lamb kidney cell line, following bovine leucosis virus (BLV) infection. 9 animals were immunised 3 times, using 300 micrograms gp51. 7 animals underwent test infection, using 2.5 x 10(3) or 2.5 x 10(4) BLV-infected lymphocytes. Serological and virological reactions of all animals, including 3 calves which had received only test infections, were followed up through 40 weeks by means of immunodiffusion test, enzyme-immuno assay, gp51 antibody radio-immuno-assay, reverse transcriptase test, syncytial test, competitive p24 radio-immuno-assay, and by transmission of whole blood in animal experiments. The results obtained from virological testing showed that 1 animal had been protected by preceding immunisation. 4 in 7 immunised and test infected animals exhibited transient BLV values, between the 7th and 16th weeks from infection. Typical leucosis infection had been induced to 2 animals. The above findings are discussed and are compared to similar results recorded by other working groups. PMID- 2559675 TI - Chrysotile A affects YAC-1 cytolytic activity of spleen cells. AB - Effect of short exposure of C57Bl/6 and F1/NZB x C57Bl/6/ mice to i.p. injected chrysotile A on YAC-1 cytolytic potential (NK cell function) of spleen cells was investigated. It was found that 3 days after injection, cytolytic activity of spleen cells was significantly abrogated and this was paralleled with an increase of alpha-naphtyl acetate esterase positive cells/macrophages in spleen cell population. In addition, effect of exposure of mice to i.p. injected chrysotile A coated with benzo(a)pyrene (BaP) was investigated. BaP did not modify the abrogative effect of chrysotile A on NK cell function. Asbestos fibres themselves were sufficient for depletion of NK cell function of spleen cells. PMID- 2559676 TI - A rare tumor at the elbow: a glomangioma. Differential diagnosis and therapy. AB - Referring to a case of a glomangioma at the elbow, several aspects of this tumor are illustrated. Since the related hemangiopericytoma can be distinguished from the glomangioma by its biological behavior, the morphological differential diagnosis of both tumors is discussed in detail. Therapeutic consequences of one or the other diagnosis are also indicated. PMID- 2559677 TI - [Paralysis of the ulnar nerve caused by osteochondromatosis]. AB - Synovial osteochondromatosis is a rare pathology (5% according to Mumenthaler, 1961) producing a compression of the ulnar nerve in the elbow and/or the wrist. The authors describe 4 cases of this pathology (3 of the elbow and 1 of the wrist) analysing its etiology and, in particular, the predisposition of the synovial membrane. PMID- 2559678 TI - [Effect of experimental laminectomy on the structure of neurons and conductive pathways of the spinal cord]. AB - In 12 dogs the effect of experimental laminectomy on the structure of motor and sensitive neurons, as well as on the structure of the conductive pathways in the spine cord has been investigated. The experimental laminectomy with a subsequent durotomy results in an increase (in 1.5-5 days of the experiment) of both hyperchromatophilic and hyperchromatophobic neurons. This demonstrates their functional reconstruction (inhibitory chromophilia, inhibited activity, fatigue and exhaustion chromatophobia). Restoration of the neuronal structure and that of the spinal cord nervous fibers takes place by the end of 10 days after the experimental laminectomy with residual foci of reversible changes in the conductive pathways system. PMID- 2559679 TI - [Neuromorphologic aspects of experimental herpetic infection]. AB - At intracerebral, intracutaneous, intracorneal and intraneural ways of infection with herpes simplex to mice and rabbits it has been stated immunomorphologically and electron microscopically that reproduction of the virus has been observed in various cells of the nervous system and accompanied with appearance of certain dystrophic changes in them. Reproductivity of the disease, duration of the incubation period, spread of the pathological process, successiveness of the lesion in the nervous system structures are determined by neurotropism of the strain and by the infection variant. The subclinical course of the infection does not exclude a productive reproduction of the virus in sensitive cells and corresponding morphologic equivalent. Inflammatory-dystrophic changes in the nervous tissue are always wider than the virus fixation zone. Reveal of the virus antigen and viral particles usually overtakes the appearance of infective allergic reactions. The electron microscopic investigations prove that a whole cycle of the virus reproduction in the nervous system cells is possible. A predominant reveal of the viral particles in the axoplasma of the nervous fibers emphasizes the importance of the axonal way for penetration of the virus into the nervous system. The complex experimental-morphological investigation is an adequate model for studying pathogenesis and pathomorphology of the herpetic infection. PMID- 2559680 TI - Neuromuscular transmission studies in human chronic Chagas' disease. AB - An electrophysiological investigation of the state of the neuromuscular transmission (nmt) was carried out in 58 patients with the diagnosis of chronic Chagas' disease. On repetitive supramaximal nerve stimulation it was found that some patients did not show abnormalities, others had decremental muscle responses, others developed enhancement of the muscle evoked potential amplitudes, while some other patients combined both types of pathological responses. The findings suggest that some patients with chronic Chagas' disease develop impairment of nmt, though data obtained in this study do not give information about neither the type of impairment nor the localization (pre or postsynaptic, or both) of the damage. PMID- 2559681 TI - Motor unit involvement in human acute Chagas' disease. AB - Thirty five patients with acute Chagas' disease who demonstrated parasitaemia at the time of the investigation were submitted to a detailed electromyographical study. With their muscles at rest, 12 patients showed fibrillation potentials and/or positive sharp waves. On volitional contraction, 7 had short duration motor unit potentials (MUPs) and low polyphasic MUPs. On motor and sensory nerve fibers conduction studies, 20 disclosed values below the lower control limit within one or more nerves. Finally, 12 patients produced a muscle decremental response on nerve supramaximal repetitive stimulation. The findings signal that primary muscle involvement, neuropathy and impairement of the neuromuscular transmission, either isolated or combined, may be found in the acute stage of human Chagas' disease. PMID- 2559682 TI - [Tumors of the brain stem: anatomo-pathologic study of 35 necropsy cases]. AB - Brain stem tumors are rare and are usually treated without histological diagnosis. Its incidence varies in literature between 1.09% and 17.5% of all cerebral tumors. The objective of our study is to present a casuistic of these tumors in 28500 necropsies performed from 1952 till 1985 at the Department of Pathology, Santa Casa Hospital, Sao Paulo. We emphasize neuropathologic aspects, and compare our cases with those of other series. Also, we point out those tumors which are most frequent in this region, with the aim of possibly contributing to a better therapeutic approach. We used some clinical data and pathological diagnosis, and localized the tumor in the brain stem. Its microscopic diagnosis was determined according to the World Health Organization criterion. Among 428 intracranial tumors observed, 35 were localized in the brain stem. Intrinsic tumors and metastasis were included, and excluded those tumors which infiltrated the brain stem. Greatest incidence occurred in the first decade and cerebral edema was the predominant cause of death. Preferential topography was in the pons and the most frequent tumor was glioblastoma multiformis (19 cases). Metastasis ranked second (9 cases), being mostly from lungs. Some authors oppose tumor biopsy in this region because of high surgical risk. We do not agree with this opinion. We believe that the diagnosis of glioma is important to rule out the characteristics of malignancy. We can not leave without considering the diagnosis of other pathologies which can cause expansion in the brain stem. Therapeutical approach can be more adequate with the histological diagnosis of glioma or of other possibilities as shown here. PMID- 2559683 TI - Lipoma of the midbrain. Post-mortem finding in a patient with breast cancer. AB - Intracranial lipomas are rare, usually do not have clinical expression and are located more frequently in the corpus callosum. Other locations include the spinal cord, midbrain tectum, superior vermis, tuber cinereum, infundibulum and more rarely cerebellopontine angle, hypothalamus, superior medullary velum and insula. We report the case of a lipoma of the left inferior colliculus which was a post-mortem finding in a woman who died of breast cancer. Although there are reports of intracranial lipomas in patients with malignant tumors there is no explanation for the co-existence of the two tumors. The present tumor also includes a segment of a nerve which is not uncommon, but a less common finding was the presence of nests of Schwann cells within it, shown by immunohistochemistry. PMID- 2559685 TI - Rotavirus vaccines: science and politics. PMID- 2559684 TI - A subpopulation of outer hair cells possessing GABA receptors with tonotopic organization. AB - The olivocochlear innervation has been postulated to regulate active mechanical processes in the mammalian cochlea. Histochemical studies led to the suggestion that a subpopulation of these efferent nerves, which predominantly terminate on outer hair cells (OHCs), are gamma-aminobutyric acid (GABA)-ergic. By means of two monoclonal antibodies, we were able to visualize GABAA-receptor immunoreactivity at the basal pole of isolated sensory cells. Both subunits of the GABAA receptor, the alpha- and beta-subunit, are known to form the transmembranous GABA/benzodiazepine-receptor complex and were present on OHCs. In addition, these inhibitory receptors were more numerous in the apical turns of the cochlea, indicating another criterion for distinguishing the apical from basal turns of the cochlea. These results support the concept that a subpopulation of axosomatic synapses at the basal pole of OHCs liberate the inhibitory neurotransmitter GABA into the synaptic cleft. Binding of the transmitter to these newly observed subsynaptic receptors is possibly followed by a change in OHC motility and a subsequent modulation of the movement of the basilar membrane. PMID- 2559686 TI - Persistent high fever and gall-bladder wall thickening in a child with primary Epstein-Barr viral infection. AB - A 3.5 year old girl presented with a history of high fever, rigors, and mild cough for 1 week. Physical examination revealed normal chest findings but gross hepatomegaly was detected. Liver function tests were abnormal and indicated biliary obstruction. Ultrasonography revealed a distended gall-bladder with increased wall thickness up to 0.6 cm. The diagnosis of primary Epstein-Barr viral infection was eventually made by specific serological study. The patient's fever subsided 2 weeks later and her liver function tests returned to normal 1 month later. Abdominal ultrasonography at this time was normal. PMID- 2559687 TI - Feline immunodeficiency virus. PMID- 2559688 TI - Avian infectious bronchitis: cross-protection studies using different Australian subtypes. AB - The cross-immunity of vaccinated chickens after challenge with some Australian infectious bronchitis viruses was assessed by humoral antibody responses and by ciliary activity in tracheal rings of vaccinated chickens following challenge. Four viruses were used for vaccination: Vac 3, Vac 4, both current infectious bronchitis vaccine viruses, and Q1/76 and N2/62. IBV N1/62 (synonym T0 and infectious bronchitis virus N9/74 (synonym Appin) were used to challenge the vaccinated chickens. Results showed a lack of correlation between humoral antibody levels and protection. Cross-immunity was found after vaccination with each subtype, but was lower for Vac 3 and Vac 4 than for Q1/76 and N2/62. PMID- 2559689 TI - Bluetongue in Australia--an entomologist's view. PMID- 2559690 TI - A brief review of studies of bovine and equine herpesviruses. PMID- 2559691 TI - Towards a vaccine against equine herpesvirus 1. PMID- 2559692 TI - An Australian perspective of foot-and-mouth disease research. PMID- 2559694 TI - Aujeszky's disease. PMID- 2559695 TI - A perspective on the pestiviruses. PMID- 2559693 TI - Herpesviruses and adenoviruses as potential vectors for vaccines in poultry. PMID- 2559696 TI - Current techniques in rapid bluetongue virus diagnosis. PMID- 2559697 TI - Practical application of nucleic acid techniques to avian disease problems. AB - A workshop in which 17 practicing scientists participated was intended to address primarily people who use or could use biotechnology in their work and was confined to five techniques. Endonuclease fingerprinting and mapping involved cleaving nucleic acid with a specific restriction enzyme and separating the nucleic acid fragments by electrophoresis. Field and vaccine isolates of Pasteurella multocida could be distinguished; Salmonella enteritidis could be divided into three groups; chlamydia could be grouped into seven groups; and vaccinia, quail pox, and fowl pox could be clearly distinguished. Preparation of nucleic acid probes involved producing large amounts of labeled oligonucleotides, usually of unknown sequence. Successful probes had been made for infectious bursal disease virus, avian influenza virus, Newcastle disease virus, and infectious bronchitis virus. In Southern, Northern, and dot blotting, either DNA or RNA fragments were placed on or transferred to a solid substrate and probed. The procedure was able to detect infectious bursal disease virus, infectious bronchitis virus, Mycoplasma gallisepticum, and Marek's disease virus. In situ hybridization involved applying a labeled probe to frozen or fixed sections or to intact cells. In Polymerase chain reaction, two primers, some distance apart, were annealed to a denatured target DNA. Repeated cycles of DNA synthesis with a thermostable polymerase, denaturing, and reannealing resulted in great amplification of a rare sequence. After 30 cycles, a rare gene sequence could be amplified more than 10(6) times. It was used successfully to detect minute quantities of influenza virus and infectious bursal disease virus, and the process was used to facilitate DNA sequencing of coccidiosis gene segments. PMID- 2559698 TI - Cellular immunity in chicks expressing fibrosarcomatous liver tumor following embryo infection with subgroup A Rous sarcoma virus. AB - Bryan standard strain of Rous sarcoma virus (BS-RSV) of subgroup A was inoculated into heavy and light breeds of chicken embryos via chorioallantoic membrane (CAM) to ascertain cell-mediated immune response, as measured by a leukocyte migration inhibition (LMI) test. Chicks hatched from eggs with pock-positive CAMs were more likely to develop liver tumors than those hatched from eggs with pock-negative CAMs. Chicks that developed tumors usually had a positive cell-mediated immune response, and those that were negative for liver tumor were negative, based on the LMI test. PMID- 2559699 TI - A primary epidemic of inclusion body hepatitis in broilers. AB - Inclusion body hepatitis (IBH) was diagnosed in 15 broiler flocks supplied by one breeder in the South Island of New Zealand. The affected flocks suffered mortality up to 30%. Malaise and slightly increased mortality were noticed by growers from about day 12 post-hatch; mortality peaked in the fourth week, and, in most flocks, declined to normally accepted levels from day 33 on. Gross signs seen at necropsy usually included bone-marrow aplasia, atrophy of the bursa of Fabricius and the thymus, and swollen hemorrhagic livers with focal necrosis. Jaundice was seen in many surviving birds. In some flocks, there was also proventricular hemorrhage, mild tracheitis, and airsacculitis. Downgrading and condemnation rates were increased in all flocks. Eosinophilic intranuclear inclusion bodies were seen in hepatocytes of some affected birds. An adenovirus was isolated from a number of cases investigated. The disease in broilers was preceded by production drops associated with feed refusal and increased mortality in the breeder stock. PMID- 2559700 TI - Comparison of egg-yolk and serum antibody titers to four avian viruses by enzyme linked immunosorbent assay using paired field samples. AB - Eggs and blood were collected from 11 hens in each of nine broiler-breeder flocks in Quebec. Serum and egg-yolk extracts were assayed for antibody titers to infectious bursal disease virus (IBDV), infectious bronchitis virus (IBV), Newcastle disease virus (NDV), and reovirus (RV) by a commercial enzyme-linked immunosorbent assay (ELISA) kit. Comparison was made between egg-yolk and serum antibody titers by a regression analysis. A high correlation was observed between serum and yolk antibody titers to all the viruses tested (r = 0.9 for IBDV, 0.84 for IBV, 0.84 for NDV, and 0.91 for RV). Antibody monitoring of commercial breeder flocks using egg yolk instead of serum with commercial ELISA plates is thus feasible and is recommended. PMID- 2559701 TI - The relationship of egg yolk and serum antibody. I. Infectious bursal disease virus. AB - Eggs and serum samples were collected from hens every 2 weeks for 50 weeks from hens caged individually so that the yolk and serum samples could be paired. It was found that the antibody titer of the yolk sample regressed significantly (F = 403.3, df = 1/135, P less than 0.001) on the titer of the serum sample. This permitted the use of the yolk titer to predict the serum titer. PMID- 2559702 TI - In situ detection by monoclonal antibody D-35.1 of cells infected with Marek's disease virus that interact with splenic ellipsoid-associated reticulum cells. AB - Immuno- and enzyme-histochemical staining procedures were used to investigate in vivo the interaction of Marek's disease virus (MDV) with splenic non-lymphoid cells. The newly developed monoclonal antibody D-35.1, which recognizes all three MDV serotypes, was used to study the localization of MDV at various times after intramuscular inoculation of 1-day-old chicks with MDV strain K. The D-35.1 positive cells were detected in the bursa of Fabricius, spleen, thymus, proventriculus, and cecal tonsils, and the number of chickens showing the cells increased between days 4 and 10. From day 21, the skin of the chickens contained D-35.1-positive feather follicles. The D-35.1 monoclonal antibody did not stain any cells in peripheral blood, nerves, kidney, and gonads at any time. In addition, D-35.1-positive cells were not detected in lymphoproliferative lesions in visceral organs and peripheral nerves. Double staining procedures on serial sections using monoclonal antibody CVI-ChNL-68.2, specific for splenic ellipsoid associated reticulum cells, revealed that the majority of D-35.1-positive cells were situated in the peri-capillary sheath of reticulum cells at day 10. The sheath of cells detected by monoclonal antibody CVI-ChNL-68.2 was disrupted, and they were clustered around D-35.1-positive cells. These results support the hypothesis that ellipsoid-associated reticulum cells are involved in the early pathogenesis of Marek's disease. PMID- 2559703 TI - Pathogenicity of Escherichia coli in aerosols for young chickens. AB - The relative pathogenicity of Esherichia coli isolates from poultry was determined by aerosol exposure of young chickens. Evidence of colisepticemia with airsacculitis and/or pericarditis and perihepatitis was evaluated. A system was devised that included the intratracheal (IT) inoculation of strain SE-17 infectious bronchitis virus (IBV) of chicks at 7 days of age followed by their aerosol exposure to E. coli culture suspensions 2 days later. Each experiment was terminated 6 days later. For comparative purposes in some studies, chicks were housed at 17 C and others at 27 C. The IBV-E. coli challenge procedure proved to be an effective way to determine the relative ability of E. coli isolates to cause death and/or gross lesions in young chickens. With some E. coli isolates, there were minimal or no obvious adverse effects from exposure except when chickens were previously inoculated with IBV. When chicks were housed at 17 C instead of 27 C, slight increases in mortality and decreases in gross lesions were generally observed, probably because the earlier deaths did not allow time for the lesions to become as evident. The E. coli isolate #18344 (Congo Red positive) was consistently more pathogenic than the Congo Red-negative version of that isolate. Cultures of E. coli previously demonstrated to be pathogenic (VA O1:K1 and DL #29) were among the most pathogenic isolates evaluated in these experiments and were similar to the Congo Red-positive #18344 isolate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559704 TI - Immunosuppressive potential and pathogenicity of a recent isolate of infectious bursal disease virus in commercial broiler chickens. AB - At 15 days of age and in the presence of measurable levels of maternal antibody against infectious bursal disease virus serotype I (1:170 virus-neutralization geometric mean titer), a recent isolate (U-28) and a prototype virulent isolate (Edgar) of the same virus caused subclinical infections in commercial broiler chickens. Isolate U-28 caused a significant reduction in the size of the bursa of Fabricius, whereas the Edgar isolate produced splenomegaly. Both isolates reduced the serological response to Newcastle disease virus. The experimental immunosuppressive potential and pathogenicity of isolate U-28 in broiler chickens confirms the role of this virus in recent infectious bursal disease outbreaks. PMID- 2559705 TI - Pathogenicity of recent isolates of infectious bursal disease virus in specific pathogen-free chickens: protection conferred by an intermediate vaccine strain. AB - The pathogenicity of recent isolates of infectious bursal disease virus and the protection conferred against them by a commercial vaccine strain of intermediate virulence were examined in specific-pathogen-free chickens. Based on clinical signs, mortality, and macroscopic lesions in susceptible chickens, the isolates designated as A-Delmarva and U-28 were distinct from a previously known serotype I virulent isolate (Edgar). Histopathological analysis of the bursa of Fabricius did not establish differences between the field isolates. Although the vaccine strain produced some degree of bursal damage in antibody-free chickens, it was significantly less severe than the damage caused by the field isolates. The active immune response induced by vaccination was cross-protective against the pathological effects produced by the different isolates used in this study. PMID- 2559706 TI - The effects of age, route of exposure, and coinfection with infectious bursal disease virus on the pathogenicity and transmissibility of chicken anemia agent (CAA). AB - Specific-pathogen-free (SPF) chickens were inoculated with several different concentrations of chicken anemia agent (CAA) by the intra-abdominal, intratracheal, or oral routes. Based on lowered hematocrit values, the birds were most susceptible to CAA introduced by the intra-abdominal route. When SPF chickens were infected with infectious bursal disease virus (IBDV) at 1 day of age, they remained susceptible to CAA up to at least 21 days, whereas birds inoculated with CAA alone were susceptible only at 1 day of age. Infectious bursal disease virus introduced at 1 day of age also increased the susceptibility of birds to contact infection with CAA and resulted in increased mortality rates in CAA inoculates. The response of SPF birds to CAA infection varied following exposure at 1 day of age to two different strains of IBDV (STC and Variant-E). Chicken anemia agent contacts and inoculates infected with the Variant-E strain were affected 1 week earlier by CAA than by STC inoculates, as evidenced by depressed hematocrits. However, the total number of birds affected was similar for both the Variant-E and STC-inoculated chickens. Commercial broiler chickens inoculated at 1, 7, 10, and 14 days of age by non-parenteral routes with CAA or a combination of CAA and IBDV had mean hematocrits that were lower than controls. Several CAA-inoculated birds were considered anemic, with hematocrit values of 25 or less, while uninoculated birds remained within normal ranges. PMID- 2559707 TI - Protection afforded infectious bronchitis virus-vaccinated sentinel chickens raised in a commercial environment. AB - The protective efficacy of three infectious bronchitis virus (IBV) vaccines for sentinel chickens raised with commercial Delmarva broiler chickens was evaluated during winter 1987. Specific-pathogen-free leghorn sentinel chickens were vaccinated with Massachusetts (Mass) alone, Mass and JMK, or Mass and Arkansas (Ark) combination live vaccines, or they remained unvaccinated. Four weeks post vaccination, sentinels were placed on broiler farms at weekly intervals for 3 weeks corresponding to weeks 4, 5, and 6 of the broiler growing cycle. Vaccine efficacy was evaluated based on IBV reisolation attempts from tracheal swabbings following a 1-week field exposure period. Sentinel chickens vaccinated with Mass and Ark combination vaccine were best protected against IBV field challenge. Only four IBV isolations were made out of a 3-week total of 36 attempts, for an 11% isolation rate. IBV vaccines containing either Mass alone or Mass and JMK offered much lower levels of protection. PMID- 2559708 TI - Hemorrhagic enteritis of turkeys: influence of maternal antibody and age at exposure. AB - The effect of maternal antibody (MAB) to hemorrhagic enteritis (HE) on the response of turkeys to infection with virulent and avirulent strains of HE virus (HEV) was examined. The influence of age at exposure and treatment with HEV antibody on development of clinical HE also was studied. MAB protected poults from clinical HE for up to 6 weeks of age. MAB also interfered with vaccination against the disease for at least 5 weeks after hatching, as indicated by absence of HEV antigen in spleens and by poor seroconversion at 6 days and at 3 weeks post-vaccination, respectively. The incidence of clinical HE in MAB-negative poults was significantly higher in poults inoculated with virus at 15 days of age or older than in poults inoculated at 1-13 days of age. Further, MAB-negative poults embryonally inoculated with virulent or avirulent strains of HEV did not develop disease; these poults developed antibody and resisted challenge with virulent virus at 6 weeks of age. Poults treated with HE antibody within 1 hour of challenge or at 1, 3, or 5 weeks before challenge with virulent virus were protected against lesions and mortality induced by HEV. These results suggest that MAB may influence susceptibility of turkeys to infection with HEV for at least 5 to 6 weeks after hatching, unlike the case with most other viral infections of poultry. The results confirm that early age resistance to clinical HE is independent of MAB and suggest that such resistance persists for up to 13 days of age. The data also suggest that turkeys lacking MAB can be immunized against HE by embryo vaccination. PMID- 2559709 TI - An outbreak of nephropathogenic H13 infectious bronchitis in commercial broilers. AB - A nephropathogenic Massachusetts strain of infectious bronchitis virus, designated H13-IB virus, was isolated from the kidneys of broilers in a commercial flock. Respiratory distress, diarrhea, depression, and high mortality were present. Gross renal lesions consisted of pale coloration, mottling, urate deposition, and swelling. The trachea contained a serous to catarrhal exudate. Microscopically, renal changes were primarily associated with the medullary region. Lesions included an interstitial mononuclear cell infiltration, urate deposition, edema, and degenerative and necrotic tubular changes. Tracheal changes included marked thickening of the mucosa, desquamation of ciliated and glandular epithelia and replacement with a stratified undifferentiated epithelium, and inflammatory cell infiltration. PMID- 2559710 TI - Hormonal contraceptives, human papillomaviruses and cervical cancer; some observations from a colposcopy clinic. AB - Observations from a colposcopy clinic from 1982-1985 are presented and show that in women with histologically proven precancerous lesions, those using hormone contraception had a significantly higher infection rate with human papillomavirus (HPV). Hormone contraception was preferred by younger women. Because information relating to sexual behaviour was not available for analysis, the findings in this study need to be interpreted with caution. The association of HPV and cervical cancers is well established. If hormonal contraceptive use and simultaneous HPV infection are associated with cervical neoplasia, and laboratory evidence supports this thesis, then further urgent investigations are needed for the implications are grave. PMID- 2559711 TI - The regional distribution of adenosine-regulating enzymes in the left and right ventricle walls of control and hypertrophic heart. AB - The transmural distribution of the adenosine-generating enzyme 5'-nucleotidase (5'N) and of the adenosine-degrading enzymes adenosine deaminase (ADA), AMP deaminase (AMP-D) and adenosine kinase (Ado-K) were determined across the walls of left and right ventricles of control and hypertrophic rat hearts. The enzyme distribution across the left ventricle wall (but not across the right wall) of normal hearts was not uniform: 5'N activity shows its highest levels in the subepicardial and in the subendocardial regions, whereas all the other enzyme activities show their lowest levels. A similar pattern of transmural distribution was also detected in other mammalian species (ox and pig). In the experimental cardiac hypertrophy, caused by two different types of chronic cardiac overload, the levels and the profiles of transmural distribution of 5'N and ADA enzyme activities may significantly change across the rat left ventricle wall. PMID- 2559712 TI - Positive regulation of the Drosophila melanogaster G6PD gene by an insertion sequence. AB - In a previous study, we have shown that the three high-G6PD activity mutants are characterized by insertion of the Ins1 sequence consisting of a core sequence flanked by two defective P elements (KP and KP'; the 32nd base of the KP was replaced by guanine in the KP') in front of exonI of the G6PD gene and that the sequence responsible for positive regulation of the G6PD gene expression might be the core sequence but not the flanking KP and KP' elements. The core sequence is composed of either one or two identical units in each mutant. In this report we present evidence (1) that insertion of the Ins1 sequence gives rise to overproduction of G6PD mRNA, (2) that the length and the 5' end of G6PD mRNA do not differ in wild-type and three mutants, (3) that the insertion site of the Ins1 sequence is the same in the mutants, and (4) that each unit of the core sequence has the same in the mutants, and (4) that each unit of the core sequence has a pair of DNase I-hypersensitive sites. The possibility exists that the binding of some regulatory proteins to the DNase I-hypersensitive sites might accelerate the transcription rate of the G6PD gene. PMID- 2559713 TI - Amplification of various esterase B's responsible for organophosphate resistance in Culex mosquitoes. PMID- 2559714 TI - Binding of two spin-labelled derivatives of chlorpromazine to human erythrocytes. AB - The binding to human intact erythrocytes of two different spin-labelled derivatives of chlorpromazine has been studied. The influence of the positively charged side chain of the drug has been the focus of our attention. The positively charged amphiphilic compound (spin derivative I) is water-soluble up to 80 microM at pH values below 5.9. The apolar analogue (spin derivative II) aggregates in aqueous buffer from the lowest concentration tested. Both spin derivatives undergo a slow reduction inside the erythrocyte. The reduced nitroxides are readily reoxidized by adding a low, non-quenching, concentration of potassium ferricyanide to the intact erythrocytes. The fractions of spin label I and II bound to the erythrocyte membrane or to the erythrocyte-extracted lipids remain constant as a function of the temperature (3-42 degrees C) and as a function of the concentration of the spin label up to 150 microM. E.s.r. spectra of both spin labels show a two-component lineshape when they are bound to intact erythrocytes. Below 35 degrees C for the positively charged spin probe, and below 32 degrees C for the apolar spin probe, the simulation of the lineshape shows that more than 50% of the spectrum originates from a slow-motion component. This slow-motion component is also found in erythrocyte-extracted lipids probed by the positively charged spin label below 25 degrees C. In contrast, no slow-motion component is detected in the range 4-40 degrees C for the apolar spin label in erythrocyte-extracted lipids. In this environment the apolar probe experiences a single fast anisotropic motion with an exponential dependence on 1/temperature. Detailed lineshape simulations take into account the exchange frequency between binding sites where the probe experiences a fast motion and binding sites where it experiences a slow motion. The exchange frequency is strongly temperature dependent. Characterization of the different motions experienced inside the different locations has been achieved and compared for whole erythrocytes and for the extracted lipids. The biochemical nature of the binding sites (membrane protein/acidic phospholipid) giving rise to the slow-motion component is discussed as a function of the polarity of the spin-labelled drug and as a function of the temperature controlling the fluidity of the lipid bulk and influencing the distribution of the drug inside the membrane. PMID- 2559715 TI - Studies on the active site of pig plasma amine oxidase. AB - Amine oxidase from pig plasma (PPAO) has two bound Cu2+ ions and at least one pyrroloquinoline quinone (PQQ) moiety as cofactors. It is shown that recovery of activity by copper-depleted PPAO is linear with respect to added Cu2+ ions. Recovery of e.s.r. and optical spectral characteristics of active-site copper parallel the recovery of catalytic activity. These results are consistent with both Cu2+ ions contributing to catalysis. Further e.s.r. studies indicate that the two copper sites in PPAO, unlike those in amine oxidases from other sources, are chemically distinct. These comparative studies establish that non-identity of the Cu2+ ions in PPAO is not a requirement for amine oxidase activity. It is shown through the use of a new assay procedure that there are two molecules of PQQ bound per molecule of protein in PPAO; only the more reactive of these PQQ moieties is required for activity. PMID- 2559716 TI - Reappraisal of the e.p.r. signals in (post)-ischaemic cardiac tissue. AB - The present study was designed to measure directly, using e.p.r. spectroscopy, oxygen-derived free radicals in (post)-ischaemic or (post)-anoxic rat hearts. Rat hearts were rapidly freeze-clamped at 77 K under normoxic, anoxic, ischaemic or reperfusion conditions. The samples were measured at three different temperatures (13, 77 and 115 K) and at several microwave power levels, and were compared with isolated rat heart mitochondria. Samples were prepared both by grinding and as tissue cuts. The two preparation techniques gave identical e.p.r. results, which excludes the occurrence of grinding artifacts. No free radical signals linked to reperfusion injury were detected. Several electron transfer centres known in the mitochondrial respiratory chain were measured. The signals previously assigned to post-ischaemic reperfusion injury were found to originate from electron transfer centres of the respiratory chain, predominantly the iron-sulphur cluster S-1 in succinate dehydrogenase. The differences in signal intensity between normoxic, ischaemic and reperfused hearts were found to result from the different redox stages of these centres under the various conditions tested. These findings do not necessarily imply that oxygen-derived free radicals are not formed in cardiac tissue during (post)-ischaemic reperfusion. The constitutive background of paramagnetism from the respiratory chain, however, seriously hampers the direct detection of comparatively low concentrations of free radicals in cardiac tissue. It is therefore expedient to focus future experiments in this field on the use of spin-trapping agents. PMID- 2559717 TI - Inactivation of skeletal-muscle UDP-glucose pyrophosphorylase by reaction with carboxylate-directed reagents. AB - Skeletal-muscle UDP-glucose pyrophosphorylase is inactivated by reaction with 2 ethoxy-N-(ethoxy-carbonyl)-1,2-dihydroquinoline (EEDQ) and 1-(3 dimethylaminopropyl-3-ethylcarbodi-imide (EDAC), two reagents specific for carboxylate groups. The former reagent is a more effective inactivator than EDAC. Although no evidence of reversible enzyme-reagent complexes of the affinity labelling type was obtained by kinetic analysis of the inactivation, the selective protection of UDP-glucose pyrophosphorylase activity against inactivation by EEDQ in the presence of uridine substrates is indicative of an active-site-directed effect. The results are consistent with the hypothesis that EEDQ modifies a single carboxylate group located in a hydrophobic domain close to the substrate-binding site, leading to enzyme inactivation. In contrast, the reaction between UDP-glucose pyrophosphorylase and EDAC appears to involve a different region of the enzyme. PMID- 2559718 TI - Hydrolysis of inositol phosphates by plant cell extracts. AB - A gel-filtered soluble fraction prepared from suspension-cultured Nicotiana tabacum cells hydrolysed inositol mono-, bis- and tris-phosphates. At a concentration of 7.5 microM the rates of hydrolysis followed the sequence Ins(1,4,5)P3 greater than Ins(1,4)P2 greater than Ins(4)P congruent to Ins(1)P. The major products of Ins(1,4,5)P3 hydrolysis identified by h.p.l.c. were Ins(1,4)P2 and Ins(4,5)P2. Ins(1,4)P2 was hydrolysed exclusively to Ins(4)P. The inclusion of Ca2+ in the incubation buffer markedly stimulated the hydrolysis of all the inositol phosphate substrates. Under identical conditions, Ca2+ inhibited the hydrolysis of inositol phosphates by soluble extracts prepared from rat brain. Half-maximal stimulation of Ins(1,4)P2 hydrolysis was obtained at free [Ca2+] of 0.6 and 1.2 microM when the Mg2+ concentration in the incubations was 0.3 and 1.0 mM respectively. This effect of Ca2+ was exerted solely by increasing the Vmax. of hydrolysis without affecting the Km for Ins(1,4)P2. Again, in contrast with brain, the hydrolysis of inositol bis- or mono-phosphates was insensitive to high concentrations of Li+. We conclude that plants contain specific Li+-insensitive inositol phosphate phosphatases that are regulated by low concentrations of Ca2+ in a manner which is different from that observed in mammalian tissues. PMID- 2559719 TI - Carnosine, homocarnosine and anserine: could they act as antioxidants in vivo? AB - Carnosine, homocarnosine and anserine have been proposed to act as antioxidants in vivo. Our studies show that all three compounds are good scavengers of the hydroxyl radical (.OH) but that none of them can react with superoxide radical, hydrogen peroxide or hypochlorous acid at biologically significant rates. None of them can bind iron ions in ways that interfere with 'site-specific' iron dependent radical damage to the sugar deoxyribose, nor can they restrict the availability of Cu2+ to phenanthroline. Homocarnosine has no effect on iron ion dependent lipid peroxidation; carnosine and anserine have weak inhibitory effects when used at high concentrations in some (but not all) assay systems. However, the ability of these compounds to interfere with a commonly used version of the thiobarbituric acid (TBA) test may have led to an overestimate of their ability to inhibit lipid peroxidation in some previous studies. By contrast, histidine stimulated iron ion-dependent lipid peroxidation. It is concluded that, because of the high concentrations present in vivo, carnosine and anserine could conceivably act as physiological antioxidants by scavenging .OH, but that they do not have a broad spectrum of antioxidant activity, and their ability to inhibit lipid peroxidation is not well established. It may be that they have a function other than antioxidant protection (e.g. buffering), but that they are safer to accumulate than histidine, which has a marked pro-oxidant action upon iron ion dependent lipid peroxidation. The inability of homocarnosine to react with HOCl, interfere with the TBA test or affect lipid peroxidation systems in the same way as carnosine is surprising in view of the apparent structural similarity between these two molecules. PMID- 2559720 TI - Neurotensin stimulates inositol trisphosphate-mediated calcium mobilization but not protein kinase C activation in HT29 cells. Involvement of a G-protein. AB - It has previously been shown that neurotensin binds to high-affinity receptors in the adenocarcinoma HT29 cell line, and that receptor occupancy leads to inositol phosphate formation. The present study was designed to investigate further the effects of neurotensin on calcium mobilization and protein kinase C (PKC) activation in HT29 cells, and to assess the role of GTP-binding proteins (G proteins) in the neurotensin response. Direct measurements of cytosolic Ca2+ variations using the fluorescent indicator quin 2 showed that neurotensin (0.1-1 microM) elicited Ca2+ transients in HT29 cells. These transients occurred after the neurotensin-stimulated formation of Ins(1,4,5)P3, as measured by means of a specific radioreceptor assay. In addition, the peptide induced a decrease in the 45Ca2+ content of cells previously equilibrated with this isotope. The peptide effect was rapid, long-lasting and concentration-dependent, with an EC50 of 2 nM. Phorbol 12-myristate 13-acetate (PMA) inhibited by 50% the neurotensin effects on both intracellular Ca2+ and inositol phosphate levels. The inhibition by PMA was abolished in PKC-depleted cells. Pertussis toxin had no effect on either the Ca2+ or inositol phosphate responses to neurotensin. Epidermal growth factor (EGF) receptors which are present in HT29 cells have been shown to be down-regulated through phosphorylation by PKC in a variety of systems. Here, PMA markedly (70 80%) inhibited EGF binding to HT29 cells. Scatchard analysis revealed that PMA abolished the high-affinity component of EGF binding, an effect that was totally reversed in PKC-depleted cells. In contrast, neurotensin slightly (10-20%) inhibited EGF binding to HT29 cells, and its effect was only partly reversed by PKC depletion. Neurotensin had no detectable effect on sn-1,2-diacylglycerol levels in HT29 cells, as measured by a specific and sensitive enzymic assay. In membranes prepared from HT29 cells, monoiodo[125I-Tyr3]neurotensin bound to a single population of receptors with a dissociation constant of 0.27 nM. Sodium and GTP inhibited neurotensin binding in a concentration-dependent manner. Maximal inhibition reached 80% with Na+ and 35% with GTP.IC50 values were 20 mM and 0.2 microM for Na+ and GTP respectively. Li+ and K+ were less effective than Na+ and the effects of GTP were shared by GDP and guanosine-5'-[beta gamma- imido]triphosphate but not by ATP. Scatchard analysis of binding data indicated that Na+ and GTP converted the high-affinity neurotensin-binding sites into lower affinity binding sites. The properties of the effects of Na+ and GTP on neurotensin-receptor interactions are characteristic of those receptors which interact with G-proteins.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2559721 TI - Staurosporine inhibits the respiratory burst and induces exocytosis in human neutrophils. AB - The protein kinase C inhibitor staurosporine influenced in different ways the functions of human neutrophils. Staurosporine prevented the enhanced protein phosphorylation in phorbol ester- and N-formylmethyionyl-leucylphenylalanine (fMLP)-stimulated cells, and was a powerful inhibitor of the respiratory burst induced by phorbol myristate acetate [IC50 (concentration causing 50% inhibition) 17 nM] and the chemotactic peptides fMLP and C5a (IC50 24 nM). It did not alter, however, the superoxide production by cell-free preparations of NADPH oxidase. Staurosporine had no effect on agonist-dependent changes in cytosolic free Ca2+ and exocytosis of specific and azurophil granules, and showed only a slight inhibition of the release of vitamin B12-binding protein induced by phorbol myristate acetate (decreased by 40% at 200 nM). On the other hand, staurosporine also exhibited neutrophil-activating properties: it induced the release of gelatinase (from secretory vesicles) and vitamin-B12-binding protein (from specific granules). These effects were protracted, concentration-dependent, insensitive to Ca2+ depletion, and strongly enhanced by cytochalasin B. Staurosporine, however, did not induce the release of beta-glucuronidase or elastase (from azurophil granules). Except for the sensitivity to cytochalasin B, these properties suggest a similarity between the exocytosis-inducing actions of staurosporine and PMA. The results obtained with staurosporine provide further evidence that different signal-transduction processes are involved in neutrophil activation, and suggest that protein phosphorylation is required for the induction of the respiratory burst, but not for exocytosis. PMID- 2559722 TI - Subfractionation of cardiac sarcolemma with wheat-germ agglutinin. AB - The properties of highly purified bovine cardiac sarcolemma subfractionated with the lectin, wheat-germ agglutinin (WGA) were studied. Two different membrane subfractions were isolated, one which was agglutinated in the presence of 1.0 mg of WGA/mg of protein (WGA+ vesicles) and a second fraction which failed to agglutinate (WGA- vesicles). These two membrane fractions had quantitatively different rates of Na+/K+-dependent, ouabain-sensitive ATPase and Na+/Ca2+ exchange activities, yet a similar protein composition, which suggests that they were both derived from the plasma membrane. WGA- vesicles had a decreased number of [3H]quinuclidinyl benzilate-binding sites and no detectable [3H]nitrendipine binding sites. Electron-microscopic and freeze-fracture analysis showed that the WGA+ fraction was composed of typical spherical sarcolemmal vesicles, whereas the WGA- fraction primarily contained elongated tubular structures suggestive of the T-tubule vesicles which were previously isolated from skeletal muscle. Assays of marker enzymes revealed that these fractions were neither sarcoplasmic reticulum nor plasma membrane from endothelial cells. Moreover, WGA agglutination did not result in the separation of right-side-out and inside-out vesicles. On the basis of these findings we propose that the WGA+ fraction corresponds to highly purified sarcolemma, whereas the WGA- fraction may be derived from T-tubule membranes. PMID- 2559723 TI - Renal effects of vanadate in rats of different ages. AB - Both in conscious and in anesthetized rats the degree of increase in renal excretion of sodium following vanadate administration is nearly the same in adult and young rats. Remarkably, in anesthetized rats i.v. administered vanadate (0.2 mg/100 g b.m.) acts distinctly longer in 20-day-old rats compared with adults. Likely, age dependent differences in inactivation of vanadate exist. The degree of inhibition of Na+/K(+)-ATPase by vanadate is lower in young than in adult rats. Vanadate has no distinct effect on renal tubular transport of p aminohippurate in rats of different ages. PMID- 2559724 TI - DNA binding protein from Streptomyces hygroscopicus: detection of binding by gel retardation, sedimentation and effects on the transcriptional activity in vitro. AB - The binding to supercoiled plasmid DNA as well as to double-stranded short DNA fragments of the DNA binding protein. ASBIII, from S. hygroscopicus has been demonstrated by gel retardation asay. As further revealed by sedimentation analysis, the protein-DNA complex formation also involves condensation of DNA and an aggregation step at higher total protein-to-DNA ratio (w/w). In vitro studies of the transcriptional activity of DNA-protein complexes showed that the ASBIII protein inhibits the overall template activity in the RNA polymerase II system to nearly similar extents for various DNA's. Preincubation of ASBIII-DNA complexes of different GC content, however, suggested a selective inhibitory effect on GC rich DNA due to the higher stability of this complex. The results also indicate that the initiation of transcription of the GC-rich DNA template is affected by the ASBIII protein. The present results on the DNA binding properties and inactivation of the transcription of the ASBIII protein suggest that this protein may be a potential candidate for the local compaction of chromosomal DNA in Streptomyces hygroscopicus. PMID- 2559725 TI - Temporally regulated expression of insulin and insulin-like growth factors and their receptors in early mammalian development. AB - Recent studies of early development in a number of invertebrate and vertebrate species have suggested that growth factors and their receptors may play important roles in differentiation as well as cell proliferation. In the mouse embryo, the expression of the receptors for insulin and insulin-like growth factors I and II (IGF-I and -II) are temporally regulated. The ontogeny of receptor and ligand expression within the insulin and IGF gene family suggests that the very earliest stages of mammalian embryogenesis may be subject to regulation by autocrine and paracrine factors from maternal and embryonic sources. PMID- 2559726 TI - Affinity labeling of ACTH receptors in bovine adrenal cortex membranes. AB - Adrenocorticotropin labeled with 125I at Tyr23 [(125I-Tyr23]ACTH) was prepared by radioiodination of ACTH1-39 followed by reverse phase HPLC purification. When incubated with bovine adrenal cortical membranes, the radioligand bound specifically to a 40-kDa membrane protein as revealed by affinity labeling. This result indicates that the bovine adrenal ACTH receptor, whose identification and characterization have proved difficult, has an Mr of about 40,000. PMID- 2559727 TI - Vitamin D3 administration increases the membrane fluidity of intestinal mitochondria. AB - The fluorescence anisotropy in the mitochondria from vitamin D-treated chicks is significantly lower than that from the vitamin D-deficient animals with the inner core probe DPH. Surface membrane fluidity, measured with the probe TMA-DPH, shows no differences between the organelles of both groups. The fluorescence studies performed in mitochondrial subfractions revealed that cholecalciferol treatment induces a decrease of lipid order parameter S (DPH) in the mitochondrial inner membrane. These results pose the question of whether vitamin D3 participates in the regulation of physiological function of the intestinal mitochondria through changes in the physical properties of the membranes. PMID- 2559728 TI - Properties of a cyclic 3'5'-nucleotide phosphodiesterase from Vigna mungo. AB - Cyclic AMP phosphodiesterase (PDE) partially purified from roots of Vigna mungo exhibited optimum activity at pH 5.5 to 6.0 and maximum enzyme activity at 50 degrees C. Levels of PDE activity in roots remained relatively constant from the first to the eleventh day after germination; on the twelfth day there was a 400% increase in PDE activity. The enzyme was stable for at least 48 hours at 28 degrees C, retaining 92% of its original activity. Plant growth hormones including gibberellic acid, indoleacetic acid and kinetin at 1.0 and 10.0 microM concentrations did not have any significant effect on enzyme activity. Nucleotides tested including cyclic 2'3' AMP, cyclic 2'3' GMP completely abolished enzyme activity at 1.0mM while cyclic 3'5' GMP, cyclic 3'5' GMP, 2'deoxy 5' ATP, 2'deoxy 5'GTP and 5'ADP were also inhibitory to the enzyme. The enzyme was stimulated by Mg2+, Fe2+ and NH4+ while Cu2+ and Fe3+ were inhibitory. Theophylline, caffeine, phosphate, pyrophosphate and EDTA were inhibitory to the enzyme. PMID- 2559729 TI - A study of chemoreception based on membrane fluidity and circular dichroism of a membrane assembly model--role of protein penetrating into the lipid bilayer. AB - We attempted to reconstitute a chemical sensing assembly by mimicking the natural constituents of cell membranes. This liposomal arrangement is able to recognize chemical stimulants by detecting perturbation of the ordered lipid bilayer due to penetration by protein molecules. It was ascertained by measuring membrane fluidity using ESR that this assembly may be able to detect individually added chemical stimulants such as short-chain-bearing odorants (isovaleric acid, isovaleraldehyde, and isoamyl alcohol etc) at a concentration of 3 x 10(-4) parts to 1 part water. This recognition mechanism may clarify both the affinity of chemical stimulants for the liposomal arrangement and the trigger action of conformational changes in poly-L-lysine (PLL) due to the penetration of the bilayer of the PLL and sodium octylsulfate complex. PMID- 2559730 TI - Fructose bisphosphatase activity in the serum of rats treated with carbon tetrachloride. AB - In this work, fructose bisphosphatase activity in the serum of rats treated by different carbon tetrachloride doses was measured. Fructose bisphosphatase activity increased very significantly with respect to the control animals in all groups assayed. The severe reduction of the activity measured in the presence of adenosine-5'-monophosphate and its stability when measured in the presence of 1-p bromotetramisole oxalate support its specific origin. These data suggest that serum FBPase activity measurement could be used as a biochemical marker in the diagnosis of hepatocellular injury. PMID- 2559731 TI - Evidence for a synaptic plasma membrane associated adenylate kinase in the rat brain. AB - About 5% of the total adenylate kinase activity in the rat forebrain was found in a subcellular fraction enriched in synaptic plasma membrane (SPM). The enzyme remained membrane bound after washing by 1M potassium acetate. It was resistant to trypsin digestion under conditions which destroyed 90% of acetylcholinesterase activity. The SPM enzyme was solubilized by 0.25% Triton X-100 resulting in a 4 fold increase in activity. Similar effects were observed when SPM was treated with phospholipases, melittin and trifluoperazine. These results suggest the occurrence of an adenylate kinase closely associated with SPM the activity of which can only be fully expressed by disturbances to the hydrophobic lipid bilayer. The enzyme can be seen as strategically located to play a role in regenerating ATP required for the manifold activities of the synaptic membrane. PMID- 2559732 TI - Chicken mitochondrial cytochrome C oxidase subunit II: comparative analysis among the vertebrates. AB - The chicken cytochrome c oxidase subunit II (COII) was cloned and sequenced. A comparison of the deduced chicken COII sequence with 4 other vertebrate counterparts revealed 64-66% amino acid sequence homology and 68-70% nucleotide sequence homology. Four peptide segments each of nine amino acids long are highly conserved across the 5 species. A redox-center was formed by three of these highly conserved domains, which include two invariant Cys and two invariant His residues for copper ion coordination, three strictly conserved Glu or Asp residues for cytochrome c binding, and highly conserved aromatic acid residues for electron transfer. PMID- 2559733 TI - Functional alpha 2-adrenoceptors in rat adipocytes: inhibition of cyclic AMP accumulation. AB - Alpha 2-adrenoceptor activation inhibits cyclic AMP accumulation in fat cells from many species. However, the presence of alpha 2-adrenoceptors in rat adipocytes has been difficult to demonstrate. We observed that alpha 2-adrenergic activation inhibits forskolin-stimulated cyclic AMP accumulation both in rat and hamster adipocytes; UK 14304, p-amino clonidine and clonidine were the agents with higher efficacy. The effect of UK 14304 was blocked by yohimbine but not by prazosin demonstrating the involvement of alpha 2-adrenoceptors. Pertussis toxin blocked the alpha 2-adrenergic effect. Our results demonstrate the presence in rat fat cells of alpha 2-adrenoceptors coupled to adenylate cyclase via "Gi". PMID- 2559734 TI - Intermediate filament-ubiquitin diseases: implications for cell sanitization. AB - The molecular pathology of chronic degenerative disease is not understood. Generally there must be two related, but opposing, processes: the direct deleterious effects of the pathogenic insult which can be chemical or viral and a cellular cytoprotective response to the insult. We have recently shown that there is a previously unsuspected link between the intracellular inclusions seen in some major chronic degenerative diseases: the inclusions contain ubiquitin immunoreactivity. The conditions include Parkinson's disease, motor neurone disease, Alzheimer's disease and alcoholic liver disease as well as astrocytomas and a myopathy. Protein ubiquitination is considered a signal for extra-lysosomal protein degradation although ubiquitin-protein conjugation may have several other important functions. Intermediate filaments are a component of some of the inclusions in diseased cells; we have previously reported that they are involved in protein sequestration for degradation by lysosomally mediated autophagy. Therefore, intermediate-filament-containing ubiquitinated inclusions may be hallmarks of cellular attempts to eliminate pathogenic insults by activating protein degradation mechanisms. Ubiquitinated inclusions could also be a hallmark of viral infections: they are in polio-virus-infected anterior horn neurones and Epstein-Barr-transformed lymphoblastoid cells. Some of the clinical observations can be reproduced experimentally in tissue culture cells. The implications of the combined clinical and experimental observations for cell sanitization and protein catabolism will be discussed. PMID- 2559735 TI - Cloning and expression of the genes for calpains and calpastatins. AB - Calpain and calpastatin are known to be very widely distributed in animal cells. Although the physiological roles of calpain have not yet been clarified, the proteinase-proteinase inhibitor system has been suggested to play important roles in various cellular functions coupled with Ca2+ mobilization. By nucleotide sequencing of the cloned cDNAs, the primary structures of calpains and calpastatins have recently been disclosed. The expression of the genes for calpains and calpastatins in vitro as well as in vivo is being studied using cDNAs and their fragments. This paper reviews several topics on the recent progress of calpain research. PMID- 2559736 TI - Control of mRNA stability during development of Dictyostelium discoideum. AB - A large group of mRNA species (which are mainly pre-spore specific) accumulate only after the formation of multicellular aggregates. They are transcribed at a constant rate from the beginning of development and their accumulation is controlled by a 10-20-fold increase in their stability. This mRNA stabilization is dependent upon multicellularity. When aggregates are dispersed, the mRNAs are destabilized; if cells are allowed to reaggregate, the destabilization is reversed. Destabilization is not due to a selective exclusion of mRNA from polyribosomes, but is a primary control event. It does not require synthesis of new RNA or protein, but it may require an interaction between ribosome and the 5' end of mRNA molecules. PMID- 2559737 TI - Effects of insulin-like growth factors on protein metabolism: why are some molecular variants more potent? AB - The insulin-like growth factors (IGFs) produce a dual anabolic effect on protein metabolism in cultured cells via a stimulus of the synthetic pathway and an inhibition of the degradative pathway. Accordingly, they offer promise as agents that may retard the extensive and life-threatening negative nitrogen balance that accompanies human polytrauma. In this report we describe the discovery of a novel, more potent, form of IGF-1, des-(1-3)-IGF-1, and explain its increased action as resulting from higher concentrations of the free peptide produced because des-(1-3)-IGF-1 binds very poorly to IGF carrier proteins released from most cell types. The truncated growth factor retains a long biological half-life because it is attached to blood IGF-binding proteins. Further understanding of the biological significance of IGF, coupled with synthetic approaches directed at the production of mutant IGF-1 molecules, can be expected to yield significant advances in the treatment of polytrauma. PMID- 2559738 TI - Lack of stereoselectivity in the inotropic and phosphodiesterase inhibitory effects of saterinone enantiomers. AB - The enantiomers of the positive inotropic and a1-adrenoceptor blocking agent saterinone (+/-)-1,2-dihydro-5-[4-[2-hydroxy-3- [4-(2-methoxyphenyl)-1 piperazinyl]propoxy] phenyl]-6-methyl-2-oxo-3-pyridine-carbonitrile, BDF 8634) have been investigated with in vitro and in vivo models in laboratory animals. In the guinea pig papillary muscle, saterinone enantiomers had equipotent inotropic activity and were also as potent as racemic saterinone; the (R)-enantiomer, however, exhibited a greater efficacy than the related compounds. Saterinone and its enantiomers were equipotent in the inhibition of phosphodiesterase PDE III activity in the guinea pig myocardium. The equipotent inotropic effects were also observed after parenteral and enteral administration in cats. In receptor binding studies, (S)-saterinone was 10-fold more potent than (R)-saterinone by inhibiting [3H]-prazosin binding to specific alpha 1-adrenoceptor sites in rat brain cortex membranes. However, in the isolated thoracic aorta of the rabbit, (S)-saterinone was only 3-fold more potent than (R)-saterinone at preventing the pressor effects of phenylephrine. When the enantiomers were tested in vivo against the pressor effects of phenylephrine in the pithed rat, (S)-saterinone was only 2-fold more potent than (R)-saterinone in its alpha 1-adrenoceptor blocking potency. Thus the enantiomers of saterinone do not display enantio-selectivity in their inotropic and PDE III inhibitory effects in vitro, nor in their cardiotonic effects in vivo. There is a slight enantio-selectivity at alpha 1-adrenoceptors in receptor binding studies, but this is reduced to biologically irrelevant magnitude in functional studies in vitro and in vivo. PMID- 2559739 TI - Effects of denopamine on rat myocardium in comparison with isoprenaline. AB - The adverse effects of denopamine (TA-064) were investigated on rat myocardium in comparison with those of isoprenaline (isoproterenol). Experiment 1. Rats were treated with isoprenaline (10 mg/kg) or denopamine (10 mg/kg) once by subcutaneous injection. 15 h after drug administration, rat hearts were isolated, and heart mitochondria were prepared. Activities of three portions of electron transport chain (NADH-cytochrome c reductase, succinate-cytochrome c reductase, and cytochrome c oxidase) of mitochondria were measured. In rats treated with isoprenaline, significant decreases in NADH-cytochrome c reductase and cytochrome c oxidase activities were observed. While, in rats treated with denopamine, NADH cytochrome c reductase activity was not decreased significantly, and decrease in cytochrome c oxidase activity was less than that observed in the isoprenaline group. Experiment 2. Rats were treated with isoprenaline (10 mg/kg) or denopamine (10 mg/kg) by subcutaneous injection once a day for 6 successive days. After the last drug administration, hearts were isolated, and cardiac membranes were prepared. Numbers of beta-adrenergic receptors were measured using 3H dihydroalprenolol. In rats treated with isoprenaline, a significant decrease in the number of beta-adrenergic receptors was observed. On the contrary, administration of denopamine did not affect significantly the number of beta adrenergic receptors. PMID- 2559740 TI - Synthesis and biological activity of dansyl thymidines. AB - The synthesis of thymidines selectively dansylated in the 3'- and 5'-positions is reported. The biological investigation showed that these fluorescent nucleosides behave as competitive inhibitors of thymidine kinase (TK) from herpes simplex virus (HSV) and are endowed with a certain degree of antiviral activity against HSV-2 and HSV-1. PMID- 2559741 TI - Influence of Epstein-Barr virus infection on B lymphocyte responses in patients with rheumatoid arthritis. AB - The influence of Epstein-Barr virus (EBV) infection on clinical and serological features of rheumatoid arthritis (RA) were studied. Patients with in-vivo activated, in-vitro spontaneously proliferating EBV-infected B lymphocytes had higher levels of serum IgG and IgA, and tended to have more extensive disease. The finding of in-vivo activated EBV-transformed B cells was not specific for RA but was also seen in patients with ankylosing spondylitis. When supernatants of spontaneously proliferating B-cell lines from patients with RA were studied, autoantibody reactivities comparable with those from patients with infectious mononucleosis were detected. These observations suggest that EBV infection might have a profound influence on B-lymphocyte responses and clinical course in patients with RA. PMID- 2559742 TI - Efficient immortalization by SV40 T DNA of skin fibroblasts from patients with Wilms' tumor associated with chromosome 11p deletion. AB - We report that transfection with a plasmid containing the SV40 early region (T) leads to a very high frequency of immortalization in two different strains of human diploid fibroblasts, each with a partial deletion of the short arm of chromosome 11. Immortalization often occurred without a recognizable "crisis" phase. Preimmortal cells showed a high frequency of spontaneous mutations and chromosomal aberrations. These results suggest that a gene involved in the control of senescence of human cells in vitro may be associated with this chromosome. PMID- 2559743 TI - Interaction of glyceraldehyde-3-phosphate dehydrogenase with AMP as studied by means of a spin-labeled analog. AB - The binding of a spin-labeled AMP analog to tetrameric glyceraldehyde-3-phosphate dehydrogenase from rabbit muscle is described. The spin label, perdeuterated and 15N-substituted 4-amino-2,2,6,6-tetramethylpiperidine-1-oxyl, was attached to C-8 of AMP (C8-SL-AMP). Up to 8 equivalents of C8-SL-AMP bind per enzyme tetramer, i.e., 2 per monomer. Combining sites are the adenine subsite of the coenzyme binding domain and the phosphate site. Glyceraldehyde 3-phosphate causes a conformational change in the enzyme that brings C8-SL-AMP molecules bound to adjacent R-axis-related subunits closer to one another by 0.2-0.3 nm and allows for spin-spin interaction between the nitroxide radicals. Similar, but less pronounced structural changes take place upon lowering the pH from 8 to 7. Addition of a single equivalent of NAD+ to a complex of the enzyme with 7.6 equivalents of C8-SL-AMP leads to the release of almost 4 C8-SL-AMP molecules. This supports our previous findings that binding of just one NAD+ molecule induces conformational changes in all four subunits. PMID- 2559744 TI - [Identification of human porins. I. Purification of a porin from human B lymphocytes (Porin 31HL) and the topochemical proof of its expression on the plasmalemma of the progenitor cell]. AB - We describe for the first time a porin (Porin 31HL) on the plasmalemm of an eukaryontic cell line, where porins have been found only on the outer mitochondrial membranes. The expression of the porin on the plasmalemm of transformed human B-lymphocytes is demonstrated by cytotoxicity- and indirect immunofluorescence techniques with living and fixed cells. The rabbit xenoantisera used were directed against purified Porin 31HL and free or acetylated synthetic peptides of its nineteen N-terminal amino acids. The three step purification procedure for Porin 31HL started from a total membrane fraction of the B-cell line, followed by ion-exchange chromatography on CM- and DEAE cellulose and a final gel filtration in SDS on Sephacryl S-300. PMID- 2559745 TI - [Identification of human porins. II. Characterization and primary structure of a 31-lDa porin from human B lymphocytes (Porin 31HL)]. AB - We characterize and describe for the first time the primary structure of a human porin with the molecular mass of 31 kDa derived from the plasmalemm of B lymphocytes (Porin 31HL). Porin 31HL is shown to be a basic, channel forming membrane protein. The protein chain is composed of 282 amino acids with a relative molecular mass of 30641 Da without derivatisation. It is not a glycoprotein. The N-terminus is acetylated. Altogether the amino-acid sequence shows 56% hydrophilic or charged amino acids arranged in alternating regions of hydrophilic or hydrophobic character as it is typical for porins. In addition the 18 N-terminal amino acids of Porin 31HL can be arranged to an amphilic alpha helix like in other porins. Porin 31HL shows approx. 29% or 24% identity to the primary structure of mitochondrial porins of Neurospora crassa and Saccharomyces cerevisiae. Partial data on mitochondrial porins from rat kidney and beef heart show sequence identity of about 90% to the human B cell porin elaborated here. PMID- 2559746 TI - A model for the complex between the helix destabilizing protein GP32 of bacteriophage T4 and single-stranded DNA. AB - A model for the structure of the complex between the helix-destabilizing protein of bacteriophage T4, GP32, and single-stranded DNA is proposed. In this model the bases are arranged in a helix, that is characterized by a relatively large distance between successive bases, a substantial base tilt, in combination with a small rotation per base. This helix is further organized into a tertiary structure, possibly a superhelix, of which the corresponding protein shell corresponds to the relatively rigid and rod-like structure that is observed in hydrodynamic experiments. It is proposed that similar structural features apply to other single-stranded DNA binding proteins in complex with polynucleotides. PMID- 2559747 TI - Theoretical modeling of DNA-monocationic lexitropsin complexation: influence of ligand binding on DNA curvature. AB - A theoretical study is presented on the complexation to DNA of a monocationic lexitropsin. Energetics and the structures of the complexes formed are analyzed for three base pair sequences of a nucleic acid octamer. The influence of the ligand binding on the nucleic acid conformation is analysed in detail. It is found that whereas the uncomplexed nucleic acid segments have very irregular structures with an overall curvature varying between 15 degrees and 20 degrees, the DNA structure becomes more regular and the curvature is strongly reduced upon the binding of a monocationic lexitropsin. PMID- 2559748 TI - Immunohistochemical evidence for the production of tumor necrosis factor by murine MH134 tumor cells as well as monocytes in tumor lesions after systemic administration of a polyalcoholized mannoglucan from Microellobosporia grisea. AB - The endogenous production of tumor necrosis factor (TNF) by tumor tissues was examined. MH134 tumor cells as well as monocytes in MH134 tumor tissues produced TNF after systemic administration of a polyalcoholized mannoglucan (MGA) from Microellobrosporia grisea as shown by the indirect immunofluorescence technique. MH134 tumor cells also produced TNF when stimulated with lipopolysaccharide in vitro. These results suggest that regression of MH134 hepatoma can be ascribed to TNF produced not only by monocytes but also by tumor cells themselves in the tumor tissue. PMID- 2559749 TI - A highly defective HIV-1 strain isolated from a healthy Gabonese individual presenting an atypical western blot. AB - In central equatorial Africa the frequency of uninterpretable or atypical Western blots (WB)--ie. antibodies to gag proteins only--can represent up to 50% of enzyme-linked immunosorbent assay (ELISA)-positive samples. To date the significance of such serology remains unknown. Nevertheless, an unusual HIV-1 strain has been isolated from the blood of a healthy Gabonese individual who presented an atypical WB. This virus, identified as isolated HIV-1OYl, grew to low titres of reverse transcriptase activity (less than 50,000 cpm/ml) and was not obviously cytopathic. Radioimmunoprecipitation and peptide ELISA studies indicated that the lack of env-specific reactivity was probably due to the absence of antibodies to the viral glycoproteins, rather than the virus encoding a highly divergent envelope protein. Molecular cloning and sequencing of the provirus proved it to be a string of HIV-1 which was genetically closer to European and North American than to African strains. Furthermore the envelope protein sequence contained all the features of a typical HIV-1 env gene. However, the tat gene derived from the proviral clone was functionally defective. Site directed mutagenesis of this gene showed that this was due to the substitution of an essential cysteine residue for a serine. Polymerase chain reaction amplification of the tat gene, as well as parts of the gag and env gene sequences of HIV-1OYl, showed that essentially all of the proviruses were defective. These data emphasize the need to view HIV isolates as populations of distinct genomes capable of complementing each other. PMID- 2559750 TI - A novel method for the purification of HIV-1 p24 protein from hybrid Ty virus like particles (Ty-VLPs). AB - The self-assembly properties of a protein encoded by the yeast retrotransposon Ty can be exploited to produce large amounts of recombinant, particulate fusion proteins as hybrid Ty virus-like particles (Ty-VLPs). This system has now been adapted to allow the release of the additional protein by incorporation of a protease cleavage site between the yeast carrier protein and the protein of interest. The purification of the additional protein is facilitated by exploiting the ease with which Ty-VLPs can be purified from other yeast cell components due to their particulate nature. We have used this modified system to produce hybrid particles containing the HIV-1 p24 protein downstream of the recognition sequence for the blood coagulation factor Xa. The p24 was released from the particles by proteolytic cleavage and rapidly separated from the residual particulate material using centrifugation and standard chromatography techniques. This procedure has been used to purify milligram quantities of HIV-1 p24 protein that reacts with anti-p24 sera and elicits the production of p24-specific antibodies in experimental animals. PMID- 2559752 TI - A lack of evidence of sexual transmission of a simian immunodeficiency agent in a semifree-ranging group of mandrills. PMID- 2559753 TI - Statistics from the World Health Organization and the Centers for Disease Control. PMID- 2559751 TI - Induction of immune deficiency syndrome in rabbits by bovine leukaemia virus. AB - Newborn rabbits were inoculated with bovine leukaemia virus (BLV). The majority of infected rabbits produced antiviral antibodies. All the seroconverted animals developed symptoms resembling AIDS and died several months after inoculation. The course of experimental infection of rabbits with BLV resembled in many respects the broad spectrum of clinical disorders associated with AIDS induced by HIV. Antibody response to virus proteins was followed by immune deficiency and signs of neuropathy, and the animals subsequently died of opportunistic infections. Virus transmission from infected babies to the mothers by contact was also observed. In some cases the virus was salvaged from lymphocytes of rabbits with the immune deficiency syndrome. The virus-specific sequences were found to be integrated at random in the DNA of haematopoietic cells and of some organs. Slight expression of viral RNAs in lymphocytes was found. Experimental infection of rabbits with BLV can be used in experiments to understand AIDS induction. PMID- 2559754 TI - Structure determination of Mengo virus. AB - The structure of Mengo virus was determined to 3.0 A resolution using human rhinovirus 14 as an initial phasing model at 8.0 A resolution. Oscillation diffraction photographs were collected at the Cornell High Energy Synchrotron Source using orthorhombic Mengo virus crystals. The crystal space group was P2(1)2(1)2(1), a = 441.4, b = 427.3 and c = 421.9 A, with one icosahedral particle per asymmetric unit, giving 60-fold noncrystallographic redundancy. The orientations of the four viral particles in the unit cell were determined with a rotation function. Their positions relative to the crystallographic symmetry axes were found by a combination of Patterson-function analysis and a subsequent R factor search using human rhinovirus 14 atomic coordinates as a model. The initial phases to 8.0 A resolution were then computed by placing human rhinovirus 14 particles in the orientations and positions of Mengo virus particles. These phases were improved by ten cycles of real-space molecular replacement averaging. Phases between 8.0 and 3.0 A resolution were obtained by molecular replacement phase extension. One or two reciprocal-space lattice points were used for each extension followed by two cycles of averaging. PMID- 2559755 TI - Linear algebraic analyses of structures with one predominant type of anomalous scatterer. AB - Further studies have been made of the information content of the exact linear equations for analyzing anomalous dispersion data in one-wavelength experiments. The case of interest concerns structures containing atoms that essentially do not scatter anomalously and one type of anomalously scattering atoms. For this case, there are three alternative ways of writing the equations. The alternative sets of equations and the transformations for transforming one set into the other are given explicitly. Comparison calculations were made with different sets of equations. Isomorphous replacement information is readily introduced into the calculations and the advantage of doing so is clearly illustrated by the results. Another aspect of the potential of the exact linear algebraic theory is its application to multiple-wavelength experiments. Successful applications of the latter have been made by several collaborative groups of investigators. PMID- 2559756 TI - Studies on regulatory mechanisms of heme biosynthesis in hepatocytes from normal and experimental-diabetic rats. Role of cAMP. AB - In the present work we have been able to demonstrate the existence of some interrelationship between intracellular level of cAMP content and phenobarbital induction of delta-aminolevulinic acid synthase, ferrochelatase, and cytochrome P 450 biosynthesis in isolated rat hepatocytes. The increase of the level of intracellular cAMP produced by activators of adenylate cyclase, inhibitors of phosphodiesterase, or added cyclic nucleotides is reflected by an increase of the phenobarbital induction effect. The greater induction observed in hepatocytes of diabetic rats may be due to a higher level of the intracellular cAMP. The lack of potentiation of added cAMP in diabetic cells is mainly due to the fact that the maximum induction that could be attained is already achieved by the effect of the preexisting high level of the endogenous cAMP. PMID- 2559757 TI - Biosynthesis of sphingomyelinase in normal and Niemann-Pick fibroblasts. AB - Deficient sphingomyelinase activity and massive storage of sphingomyelin are common to two clinically different forms of Niemann-Pick disease, called types A and B. Polyclonal antisera to human sphingomyelinase precipitated both enzyme activity and the polypeptide chain of purified placental sphingomyelinase. In normal fibroblasts, following a 19-h labelling period with [35S]methionine and immunoprecipitation of the labelled proteins, sphingomyelinase occurred as a single polypeptide with a mean molecular mass of 110 kilodaltons (kDa). Niemann Pick disease type A and B fibroblasts also synthesized a sphingomyelinase polypeptide having the same molecular mass as that found in normal fibroblasts. In I-cell disease fibroblasts, a reduced amount of cross-reacting material was detected, suggesting that sphingomyelinase may be targeted to the lysosome via the phosphomannosyl receptor. Pulse-chase experiments demonstrated sphingomyelinase processing, as judged by a substantial loss of radiolabel and the appearance of an 84-kDa intermediate form of the enzyme. These results confirm and extend previous work based on autopsy specimens and urine, and show that Niemann-Pick disease fibroblasts synthesize a sphingomyelinase polypeptide. We show for the first time that an 84-kDa processed form of the enzyme is biosynthetically related to the 110-kDa polypeptide. PMID- 2559758 TI - Glutamate receptor subtypes may be classified into two major categories: a study on Xenopus oocytes injected with rat brain mRNA. AB - Three major subtypes of glutamate receptors that are coupled to cation channels are known. Recently an additional subtype that is coupled to G proteins and stimulates inositol phospholipid metabolism (the metabotropic glutamate receptor) has been proposed. The pharmacological characteristics of this receptor have now been examined. Although it shares some agonists with N-methyl-D-aspartate- and quisqualate-subtype receptors, it shares virtually no antagonists with any of the three cation channel-coupled receptor subtypes. Thus the metabotropic glutamate receptor belongs to a receptor category that is completely different from that of the other three receptor subtypes, not only functionally, but also pharmacologically. PMID- 2559759 TI - Neural cell adhesion molecules influence second messenger systems. AB - We have investigated the influence of the neural cell adhesion molecules L1 and N CAM on second messenger systems using a PC12 rat pheochromocytoma cell line as a model and triggering cell surface receptors by specific antibody binding. Antibodies directed against L1 and N-CAM, but not against other cell surface components, reduce intracellular levels of the inositol phosphates IP2 and IP3, while intracellular levels of cAMP are unaffected. Antibodies against L1 and N CAM also reduce intracellular pH and increase intracellular Ca2+ by opening Ca2+ channels in a pertussis toxin-inhibitable manner, suggesting the involvement of a G protein in the signal transduction process. Cross-linking of the adhesion molecules on the surface membrane is not required for the effects to occur. Furthermore, adhesion of single PC12 cells to each other elicits effects on intracellular pH and Ca2+ similar to those seen after application, underscoring the physiological significance of the observed changes. PMID- 2559760 TI - Primary structure and functional expression of a mammalian skeletal muscle sodium channel. AB - We describe the isolation and characterization of a cDNA encoding the alpha subunit of a new voltage-sensitive sodium channel, microI, from rat skeletal muscle. The 1840 amino acid microI peptide is homologous to alpha subunits from rat brain, but, like the protein from eel electroplax, lacks an extended (approximately 200) amino acid segment between homologous domains I and II. Northern blot analysis indicates that the 8.5 kb microI transcript is preferentially expressed in skeletal muscle. Sodium channels expressed in Xenopus oocytes from synthetic RNA encoding microI are blocked by tetrodotoxin and mu conotoxin at concentrations near 5 nM. The expressed sodium channels have gating kinetics similar to the native channels in rat muscle fibers, except that inactivation occurs more slowly. PMID- 2559761 TI - Regulation of phosphatidylcholine biosynthesis in activated alveolar type II cells. AB - The biosynthesis of phosphatidylcholine was studied in a population of activated Type II cells isolated from the lungs of rats treated with silica. Type II cells were separated by centrifugal elutriation into two populations, designated Type IIA and Type IIB. The Type IIB or activated population consisted of Type II cells that were larger than normal cells; Type IIA cells were morphologically similar to normal Type II cells. Type IIB cells incorporated more [Me-14C]choline into both total phosphatidylcholine and disaturated phosphatidylcholine than did Type IIA or control Type II cells. Measurement of the pool sizes of the choline containing precursors to phosphatidylcholine indicated that the biosynthesis of phosphatidylcholine was increased 4- to 5-fold in Type IIB cells. Increased conversion of cholinephosphate to CDP-choline was associated with increased phosphatidylcholine biosynthesis in Type IIB cells. Cholinephosphate cytidylyltransferase activity was increased approximately threefold in Type IIB cells. Subcellular fractionation indicated that essentially all of the increase in cytidylyltransferase activity was associated with the particulate fraction (100,000 x g pellet). In Type IIB cells, the particulate fraction contained 83% of the total cellular cytidylyltransferase activity; in control cells, this fraction contained 67% of the total activity. The specific activity of the cytidylyltransferase associated with the particulate fraction was increased twofold in Type IIB cells. The specific activity of the cytosolic enzyme was similar to that in control cells. Cholinephosphotransferase specific activity was increased approximately twofold in the activated Type II cells. The specific activity of choline kinase was the same as that in control Type II cells. These results demonstrate that the increased biosynthesis of phosphatidylcholine in Type IIB cells is a result of stimulation of the CDP-choline pathway. This study indicates that both cholinephosphate cytidylyltransferase and cholinephosphotransferase may be involved in regulating the de novo biosynthesis of phosphatidylcholine in alveolar Type II cells. PMID- 2559762 TI - Mapping the functional domains of the beta-adrenergic receptor. AB - The beta-adrenergic receptor (beta AR) serves as a model system for analysis of the structure-function relationships of G-protein-coupled receptors. Genetic analysis of the beta AR has demonstrated that the ligand-binding domain of this protein lies within the hydrophobic putative transmembrane core, involving specific amino acid residues in several of the transmembrane helices of the receptor. Site-directed mutagenesis of the receptor in conjunction with structural alterations of the ligands has revealed specific molecular interactions that are important for recognition of the ligand by the receptor. In addition, cytoplasmically exposed regions of the beta AR that are required for the activation of Gs have been identified. Because of the structural similarities among G-protein-coupled receptors, information gained from genetic analysis of the beta AR should prove useful in the development of specific agonists and antagonists for other receptors of this class. PMID- 2559763 TI - Expression of a prokaryotic gene in cultured lung endothelial cells after lipofection with a plasmid vector. AB - Transfection of cultured cells with functioning foreign genes can be a powerful tool for delineating mechanisms controlling gene expression and for manipulating cellular synthetic machinery. We transfected cultured bovine pulmonary artery endothelial cells with a plasmid (pRSVCAT) containing the prokaryotic gene, chloramphenicol acetyltransferase (CAT), driven by a Rous sarcoma virus (RSV) promoter. Transfection was accomplished by binding the plasmid DNA to specially synthesized cationic liposomes and incubating cell monolayers with the liposome DNA complex (a process called lipofection). We found marked CAT expression in endothelial cells by 24 h after lipofection (complete chloramphenicol acetylation in a 4-h assay incubation with less than 0.1 mg cell protein). Marked CAT expression persisted after splitting the cells 1:2 at 6 days after lipofection. Detectable CAT activity was present 14 days after lipofection following two 1:2 splits of the cells. CAT expression was related to the quantity of plasmid DNA used for lipofection; 1.0 micrograms DNA per 60-mm dish of cells resulted in less CAT activity at 48 h than did 5 or 10 micrograms DNA per dish. Ten micrograms of DNA-liposome complex per dish caused substantial numbers of endothelial cells to detach from the dish, but little cytotoxic effect was seen with 5 or 1 micrograms DNA-liposome complex per dish. This simple, highly efficient method for introducing functioning foreign genes into cultured endothelial cells will permit a broad range of studies of molecular mechanisms of endothelial cell function and studies of the consequences of highly specific modifications in protein synthesis on endothelial responses. PMID- 2559764 TI - [Intracytoplasmic eosinophilic inclusions in dorsal root ganglia and spinal nerve roots from an autopsy case of unusual familial ataxia with cerebrospinal fluid abnormality]. AB - Studies were performed on the light and electron microscopic structures of the dorsal root ganglia (DRG) and spinal nerve roots of the 4th lumbar nerve obtained by autopsy from a 49-year-old man with unusual familial ataxia, who showed varied neurological manifestations such as progressive ataxia, action tremors, pyramidal tract signs, mild deep sensory disturbances and autonomic dysfunctions during a 30-year period of illness, and had 2 siblings, one male and female, similarly affected and close consanguineous marriages in his family. On laboratory examinations, blood chemistry disclosed no significant findings. Repeated spinal taps showed constant xanthochromia and elevated protein in the cerebrospinal fluids. A PEG and cranial CTs revealed a progressive brain atrophy. NCVs and EMGs in the extremities were within normal limits. There was no chromosomal abnormality. Light microscopically, intracytoplasmic eosinophilic inclusions (IEIs) with pale rim, which showed varied sizes and rounded shapes, occurred within neurons in the DRG, particularly in small neurons. Many of the small neurons had numerous IEIs, and several rounded granules with a high degree of eosinophilia, measuring below 5 microns in diameter. Generally the small neurons showed atrophic, while most large neurons showed no remarkable change although they had a small number of IEIs and granules located in the perikaryal periphery. Most satellite cells, and some Schwann cells in the DRG, ventral and dorsal roots had IEIs similar to those seen in the neurons. No IEIs occurred intraaxonally, and there was seen no degenerative process in the DRG and roots except a connective tissue fiber proliferation in the DRG.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559765 TI - [A quantitative evaluation of brain computerized tomography in children using color image analyzer]. AB - We attempted the quantitative analysis of brain computerized tomographic scans in children using Color Image Analyzer. A consecutive series of 167 computerized tomographic scans were reviewed. Areas of subarachnoid spaces, cavums, ventricles and cerebellums were measured on three slices: A slice is at the level of head of caudate nucleus, anterior horn of lateral ventricle and third ventricle. B slice is at the level of body of lateral ventricle. C slice is at the level of sella turcica and pons. We investigated these values compared with Evans ratio, Cella Media Index, cerebellar atrophy score and visually evaluations. Serial brain CT scans of eight patient with infantile spasms were also evaluated for the assessment of the brain shrinkage after ACTH therapy. 1) The ratios of the subarachnoid space/the intracranial area on A and B slices (SAS A%, SAS B%) were significantly higher in the patients of severe brain atrophy. 2) There were linear relationship between Evans ratio and SAS A% (r = 0.405, p less than 0.001), Cella Media Index and the ratio of the lateral ventricles/the intracranial areas on B slice (r = -0.501, p less than 0.001), and the cerebellar atrophy score by Une and SAS C% (r = 0.369, p less than 0.001). 3) In the normal patients, the values of SAS A% and SAS B% were much greater in less than 1.5 years old children. These results suggest that the trend of CT findings related to age may reflect physiological changes of the space between the skull and the brain with age. 4) Brain shrinkage after ACTH therapy was more pronounced in the subarachnoid space than the ventricle.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559767 TI - [Expression of insulin-like growth factor I (IGF-I) and epidermal growth factor (EGF) receptors in primary non-glial human brain tumors]. AB - The expression of IGF-I and EGF receptors in the primary non-glial brain tumors (8 meningiomas, 2 neurinomas, 1 hemangioblastoma, 2 primary malignant lymphomas) was analyzed by using in vitro quantitative autoradiographic techniques. Specific binding sites for IGF-I were co-localized with those for EGF in the meningiomas and the hemangioblastoma examined. However, in the neurinomas and the malignant lymphomas, only IGF-I binding sites were present. In addition, IGF-I and EGF synergistically increased 3H-thymidine incorporation into DNA synthesis by the primary cultured meningioma cells, in dose-dependent manner. These observations can be interpreted to mean that both IGF-I and EGF may exist as autocrine or paracrine peptides involved in the growth not only of glioma but also of non glial brain tumors. PMID- 2559766 TI - [Role of perivascular phagocyte (FGP) of cerebral small blood vessel in cerebral edema]. AB - Small cerebral blood vessels are provided with slender phagocytic cells which were designated as FGP (fluorescent granular perithelial) cells from their localization and property. They were derived from pial cells and became potent in the uptake capacity for exogenous substance about at one week after birth. The present study aims to confirm the role of FGP cells and to clarify morphological changes of them in cerebral edema. Cerebral cortices of Wistar rats received head injury (caused with dry ice) were studied electron microscopically at 0.5, 2, 5, 10, 16, 30, 48 and 72 hours after the injury. At the same interval, HRP (horseradish peroxidase) was injected intravenously for the demonstration of uptake capacity of FGP cells. The authors' attention focused on the change of ultrastructure of vascular wall including perivascular phagocytic FGP cell around small blood vessels and on the uptake capacity of FGP cells at each time. From 2 to 5 hours after the cold injury, pinocytotic vesicles often appeared in endothelium and smooth muscle cells, and blood plasma infiltrated into the perivascular space and was taken in the FGP cells. The FGP cells in control were provided with intense round inclusion bodies and looked moderately intense. However, the FGP cells in the experimental animals after the injury became swollen and vacuolized following the fusion of inclusion bodies and breakdown of their limiting membrane and looked pale owing to presence of vacuoles. Under this condition, the uptake capacity of FGP cells for HRP decreased remarkably, and ACPase which was confined to the inclusion bodies in controls spread diffusely into cytoplasm.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559768 TI - Hepatoblastoma: successful resection in an adult. PMID- 2559769 TI - NMR distance measurements in DNA duplexes: sugars and bases have the same correlation times. AB - To evaluate whether the sugar moieties of short DNA duplexes exhibit local motion of sufficient amplitude to affect interproton distance measurements, we have carried out a series of time-dependent NOESY experiments at increasingly shorter mixing times on dodecamer DNA duplexes. By use of the cytosine H5-H6 vector as a known distance in the bases and the geminal 2'H-2''H vector as a known distance in the sugars, the corresponding apparent cross-relaxation rates were sampled at various mixing times. While the ratio of the inverse sixth power of these two fixed distances is in the range 6-7, when the system is sampled at 100 ms the apparent initial rate of growth of the 2'H-2''H NOESY crosspeak is only 1.9-2.0 times faster than that of the H5-H6 crosspeak--in agreement with the results of Clore and Gronenborn [Clore, G. M., & Gronenborn, A. M. (1984) FEBS Lett. 172, 219; (1984) FEBS Lett. 175, 117] and of Gronenborn and Clore [Gronenborn, A. M., & Clore, G. M. (1985) Prog. NMR Spectrosc. 17, 1]. This observation was interpreted to indicate the existence of internal mobility with a 3-fold shorter correlation time for the sugar moieties in DNA and led to the use of this shorter correlation time to estimate sugar-sugar proton distances and many sugar-base proton distances in subsequent DNA structure determination. We have examined 2'H 2"H cross-relaxation and H5-H6 cross-relaxation at 100, 90, 60, 30, and 15 ms in dodecamer DNA duplexes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559770 TI - Synthesis and properties of azidonitrophenyl pyrophosphate, a photoaffinity probe of the nucleotide binding sites of mitochondrial F1-ATPase. AB - 4-Azido-2-nitrophenyl pyrophosphate (azido-PPi) labeled with 32P in the alpha position was prepared and used to photolabel beef heart mitochondrial F1. Azido PPi was hydrolyzed by yeast inorganic pyrophosphatase, but not by mitochondrial F1-ATPase. Incubation of F1 with [alpha-32P]azido-PPi in the dark under conditions of saturation resulted in the binding of the photoprobe to three sites, two of which exhibited a high affinity (Kd = 2 microM), the third one having a lower affinity (Kd = 300 microM). Mg2+ was required for binding. As with PPi [Issartel et al. (1987) J. Biol. Chem. 262, 13538-13544], the binding of 3 mol of azido-PPi/mol of F1 resulted in the release of one tightly bound nucleotide. ADP, AMP-PNP, and PPi competed with azido-PPi for binding to F1, but Pi and the phosphate analogue azidonitrophenyl phosphate did not. The binding of [32P]Pi to F1 was enhanced at low concentrations of azido-PPi, as it was in the presence of low concentrations of PPi. Sulfite, which is thought to bind to an anion-binding site on F1, inhibited competitively the binding of both ADP and azido-PPi, suggesting that the postulated anion-binding site of F1 is related to the exchangeable nucleotide-binding sites. Upon photoirradiation of F1 in the presence of [alpha-32P]azido-PPi, the photoprobe became covalently bound with concomitant inactivation of F1. The plots relating the inactivation of F1 to the covalent binding of the probe were rectilinear up to 50% inactivation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559771 TI - Antibody-nucleic acid complexes. Oligo(dG)n and -(dT)n specificities associated with anti-DNA antibodies from autoimmune MRL mice. AB - The specificity of anti-DNA antibodies in the sera of unimmunized autoimmune MRL mice was initially assessed via an enzyme-linked immunosorbent assay (ELISA). Antibody binding profiles to a panel of immobilized antigens (AMP-, GMP-, CMP-, UMP-, and TMP-BSA, ss- and dsDNA) demonstrated high levels of immunoglobulins reacting with GMP and ssDNA and intermediate levels with AMP, TMP, and dsDNA. Fractionation of serum anti-DNA antibodies into subsets on the basis of their binding to GMP- and TMP-agarose indicated that the resulting GMP- or TMP-reactive antibodies bound to their homologous nucleotides and ssDNA. Competition inhibition studies with soluble mono-, oligo-, and polynucleotides revealed that GMP- and TMP-reactive antibodies were highly specific for oligo(dG)n and -(dT)n sequences, respectively. Whereas the relative affinity of TMP-reactive autoantibodies to oligo(dT)n increased with oligonucleotide length (n = 2, 4, 6, 8, 10, 15), GMP-reactive antibodies preferentially recognized oligo(dG)10 (Ka congruent to 1 x 10(7) M-1). While neither antibody recognized oligo(dA)8 and (dC)8 competitors, mixed-base oligonucleotides were inhibitory at concentrations approximately 10-fold greater than similarly sized oligo(dG)n and -(dT)n sequences. Similar characterizations of both pooled and individual MRL sera indicated that anti-DNA antibodies represent 8-10% of the total serum IgG. More importantly, GMP-reactive autoantibodies predominated and accounted for 60-70% of the entire unbound anti-DNA antibody population. PMID- 2559772 TI - Coenzyme F430 as a possible catalyst for the reductive dehalogenation of chlorinated C1 hydrocarbons in methanogenic bacteria. AB - Corrinoids, such as aquocobalamin, methylcobalamin, and (cyanoaquo)cobinamide, catalyze the reductive dehalogenation of CCl4 with titanium(III) citrate as the electron donor [Krone et al. (1989) Biochemistry 28, 4908-4914]. We report here that this reaction is also effectively mediated by the nickel-containing porphinoid, coenzyme F430, found in methanogenic bacteria. Chloroform, methylene chloride, methyl chloride, and methane were detected as intermediates and products. Ethane was formed in trace amounts, and several as yet unidentified nonvolatile compounds were also generated. The rate of dehalogenation decreased in the series of CCl4, CHCl3, and CH2Cl2. With coenzyme F430 as the catalyst, the reduction of CH3Cl to CH4 proceeded more than 50 times faster than with aquocobalamin. Cell suspensions of Methanosarcina barkeri were found to catalyze the reductive dehalogenation of CCl4 with CO as the electron donor (E'0 = -0.524 V). Methylene chloride was the main end product. The kinetics of CHCl3 and CH2Cl2 formation from CCl4 were similar to those with coenzyme F430 or aquocobalamin as catalysts and titanium(III) citrate as the reductant. PMID- 2559773 TI - Photodissociable endogenous ligand in alkaline-reduced cytochrome c peroxidase implicates distal protein tension. AB - Laser excitation of alkaline- (pH 8.5) reduced cytochrome c peroxidase (CCP) produces resonance Raman (RR) bands arising from both low- and high-spin heme species (nu 3 = 1493/1471 cm-1) even though in the absence of laser excitation the absorption spectrum is characteristic of a purely low-spin species. The high spin fraction is higher in a stationary than in a rotating sample, indicating that the high-spin contribution arises from photolysis induced by the Raman laser. This conclusion was confirmed by monitoring the absorption spectrum during laser irradiation. Photolability of the low-spin form is somewhat less than that of the CO adduct. The endogenous photolabile ligand is proposed to be the distal histidine residue, His-52. Recent picosecond absorption measurements (Jongeward et al., 1988) show that imidazole ligands in heme proteins do photodissociate but recombine in picoseconds, leading to net photostability on longer time scales. It is proposed that a fraction of the His-52 residues recombine much more slowly in CCP because of protein strain in the ligated form. This strain can also explain the anomalously rapid rate of CO binding to alkaline CCP. PMID- 2559774 TI - Purification and characterization of bovine heart phosphoinositide-specific phospholipase C: kinetic analysis of the Ca2+ requirement and La3+ inhibition. AB - Bovine heart contains multiple phosphoinositide-specific phospholipase C (PIC) activities separable by ion-exchange chromatography. One PIC activity was purified to apparent homogeneity and migrated as a single band of Mr 85,000 on SDS-PAGE. The purified PIC was characterized with sonicated suspensions of either pure phosphatidylinositol 4,5-bisphosphate (PIP2) or phosphatidylinositol (PI) as substrates. At pH 7, apparent Vmax and Km values were higher for PIP2 than for PI, but the value of Vmax/Km was similar for the two substrates. PIC required Ca2+ for the hydrolysis of either PI or PIP2, and increasing free Ca2+ concentrations from 20 to 300 nM saturated PIC activity. The requirement of Ca2+ for PIC activity and the sensitivity of PIC to Ca2+ concentrations in the physiological range suggested the ion may be a cofactor. The PIC reaction mechanism was determined by two-substrate kinetic analysis; the data fit a model in which PIC contained single sites for Ca2+ and phosphoinositide, and utilized a rapid-equilibrium, random-order ternary mechanism for phosphoinositide hydrolysis. The KCa value for either PI or PIP2 hydrolysis was approximately 30 nM, suggesting resting intracellular free Ca2+ concentrations are sufficient to saturate the Ca2+ site of PIC. La3+ was used as a calcium analogue to modulate PIC activity. Low concentrations of LaCl3 (0.01-0.3 microM) inhibited PIC activity competitively with respect to calcium, consistent with a Ca2+ binding site on the enzyme. PMID- 2559775 TI - Probing the hydration of the minor groove of A.T synthetic DNA polymers by volume and heat changes. AB - The minor-groove ligand netropsin provides a sensitive probe of the hydration difference between poly(dA).poly(dT) and poly[d(AT)].poly[d(AT)]. We have measured the volume change delta V accompanying binding of netropsin to these polymers, using an improved magnetic suspension densimeter. For poly(dA).poly(dT) we find delta V = +97 mL/mol of bound netropsin at pH 7.0 and 10 mM sodium phosphate buffer. For poly[d(AT)].poly[d(AT)] we find delta V = -16 mL/mol of bound netropsin. This striking differential effect suggests that the poly(dA).poly(dT) duplex compresses more water (or is more extensively hydrated). From our enthalpy and entropy results we estimate the approximately 10 water molecules, immobilized in the minor groove of this system, are displaced by each netropsin bound. The volume increase, however, is substantially larger than can be explained by a simple melting of these immobilized water molecules in the minor groove. A decompression of at least 40 water molecules must attend the complexation to the poly(dA).poly(dT) duplex. This suggests that the conformation change attending the binding of the drug to this polymer duplex causes a further dehydration, whereas no such change in dehydration and configuration for the heteropolymer system is indicated. PMID- 2559776 TI - Spin-label ESR of bacteriophage M13 coat protein in mixed lipid bilayers. Characterization of molecular selectivity of charged phospholipids for the bacteriophage M13 coat protein in lipid bilayers. AB - Bacteriophage M13 major coat protein has been incorporated at different lipid/protein ratios in lipid bilayers consisting of various ratios of dimyristoylphosphatidylcholine (DMPC) to dimyristoylphosphatidylglycerol (DMPG). Spin-label ESR experiments were performed with phospholipids labeled at the C-14 position of the sn-2 chain. For M13 coat protein recombinants with DMPC alone, the relative association constants were determined for the phosphatidylcholine, phosphatidylglycerol, and phosphatidic acid spin-labels and found to be 1.0, 1.0, and 2.1 relative to the background DMPC, respectively. The number of association sites for each phospholipid on the protein was found to be 4 per protein monomer. The intrinsic off-rates for lipid exchange at the intramembranous surface of the protein in DMPC alone at 30 degrees C were found to be 5 X 10(6), 6 X 10(6), and 2 X 10(6) s-1 for the phosphatidylcholine, phosphatidylglycerol, and phosphatidic acid spin-labels, respectively. Adding DMPG to the DMPC lipid system increased the exchange rates of the lipids on and off the protein. By gel filtration chromatography, it is found that protein aggregation is reduced after addition of DMPG to the lipid system. This is in agreement with measurements of tryptophan fluorescence, which show a decrease in quenching efficiency after introduction of DMPG in the lipid system. The results are interpreted in terms of a model relating the ESR data to the size of the protein-lipid aggregates. PMID- 2559777 TI - The gp120-CD4 interface: structural, immunological and pathological considerations. PMID- 2559779 TI - Luteotropic and luteolytic responsiveness of ovine luteal cells in long-term culture. AB - Ovine luteal cells were collected and plated 36 h (Day 2) after injection of human chorionic gonadotropin (Day 0) to induce ovulation. Cells were maintained (Days 2-12) in Medium 199 containing 5% calf serum, which was replaced daily. Progesterone secretion was not stimulated (p greater than 0.05) by luteinizing hormone (LH, 10 ng/ml or 100 ng/ml) at any time during culture. However, it was enhanced (p less than 0.05) with a 24-h pulse of dibutyryl adenosine 3', 5' monophosphate (dbcAMP) during early (2.2-fold stimulation over basal; Days 5,6) or mid- (1.7-fold stimulation over basal: Days 8,9) culture if the pulsing medium contained serum, but not if serum had been withdrawn for 24 h. Continuous exposure of cultures to dbcAMP (2 mM, Days 3-12) resulted in continuously stimulated (p less than 0.05) progesterone secretion (range 1.8- to 4.1-fold stimulation). An increased (p less than 0.05) percentage of cells staining positive for 3 beta-hydroxy-delta 5-steroid dehydrogenase-delta 5, delta 4 isomerase (3 beta HSD) activity were recovered on Day 12 in cultures incubated (Days 3-12) with dbcAMP. Incubation of cultures continuously with prostaglandin F2 alpha (PGF2 alpha) produced dose-dependent inhibition (p less than 0.05) of progesterone secretion. Reduced numbers of 3 beta HSD-positive cells were recovered from these incubations. These experiments demonstrate luteotropic (dbcAMP) as well as luteolytic (PGF2 alpha) effects on ovine luteal cells in long term culture. This study provides evidence that these cultures will be useful for investigating the development of hormonal regulation of luteal function. PMID- 2559778 TI - Characterization of platelet-activating factor binding sites on uterine membranes from pregnant rabbits. AB - One of the earliest signs of endometrial preparation for blastocyst implantation is a localized increase in capillary permeability, an event that is essentially inflammatory in character and thought to be a prerequisite for subsequent decidual tissue formation. Platelet-activating factor (PAF), chemically identified as 1-O-alkyl-2-acetyl-sn-glycero-3-phosphorylcholine, is a very potent vasoactive compound that recently has been implicated in the implantation process. In the present study, PAF binding sites are characterized in the rabbit uterus. A specific, reversible, saturable, and thermally labile binding of [3H]PAF to uterine membranes has been demonstrated, exhibiting multiple binding sites. The equilibrium dissociation constant (Kd) of the higher affinity binding site (type 1) was 3.6 +/- 0.4 nM (mean +/- SD) with a binding capacity (Bmax) of 3.4 +/- 1.6 pmol/mg protein. The second (lower affinity) binding site (type 2) had an apparent Kd of 114.6 +/- 13.5 nM and a Bmax of 164.3 +/- 17.6 pmol/mg membrane protein, under the conditions of maximal [3H]PAF binding, 25 degrees C, 150 min. Incubations at 4 degrees C for up to 3 h yielded only 30% of the Bmax observed at 25 degrees C. In crude and purified endometrial membrane preparations in which the PAF binding was predominantly located, the affinity of the binding for PAF was significantly higher than for the whole uterus, giving Kds of 1.5 +/- 0.8 and 0.8 +/- 0.5 nM; these latter values were not significantly different. However, the Bmax values of 3.9 +/- 0.9 pmol/mg protein and 376.8 +/- 163.3 fmol/mg protein for the two endometrial preparations, respectively, did differ significantly. Kinetic analysis at 25 degrees C resulted in a calculated Kd of 3.28 +/- 1.14 nM, which did not differ from the value for for the whole uterus at the same temperature, but was greater than for the endometrial preparations. Using 4 nM [3H]PAF to selectively label only the type 1 binding sites, the relative potencies of PAF and its antagonists in displacing [3H]PAF were lyso-PAF greater than CV3988 greater than PAF greater than U66985 greater than A02405 greater than BN52021 greater than U66982. The antagonists SRI 63,441 and L652,731 were ineffective in displacing [3H]PAF at up to 5000-fold molar excess of [3H]PAF. [3H]Lyso-PAF binding at 4 nM was displaceable by PAF. All cations tested, i.e. Ca2+, Mg2+, K+, Na+, and Li+, inhibited [3H]PAF binding. Serine hydrolase inhibitors, diisopropylfluorophosphate (DFP) and phenylmethylsulfonyl fluoride (PMSF), inhibited binding, but bacitracin, leupeptin, and antipain stabilized it.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2559780 TI - Effect of temperature on alpha-adrenoceptor affinity and contractility of rabbit ear blood vessels. AB - We have studied contractile responses to norepinephrine (NE) and K+ of isolated rabbit ear arteries and veins in an effort to determine how alpha-adrenoceptor affinity and smooth muscle contractility affect responsiveness at different temperatures. Arteries have predominantly alpha 1-adrenoceptors and veins have predominantly receptors of the alpha 2-type. We obtained dose-response curves to NE at 9, 16, 23, 30, 37 and 42 degrees C in the absence and presence of the irreversible alpha-adrenoceptor blocker phenoxybenzamine (POB). In both vessels, affinity of the alpha-adrenoceptors was determined by comparing equieffective doses of NE before and after blockade of the receptors. In other experiments, we obtained contractions to 80 mM KCl at the same temperatures. Affinity of the ear arterial alpha 1-receptors was maximal at 23 degrees C, while affinity of ear venous alpha 2-receptors increased linearly with cooling to 9 degrees C. Responses to K+ in both vessels decreased with cooling. Taken together, the changes in receptor affinity and response to K+ may explain the shape of curves relating contractile strength to temperature. PMID- 2559781 TI - [Multifactor resistance to various antineoplastic agents]. PMID- 2559782 TI - Thin layer chromatography of dansyl and dinitrophenyl derivatives of amino acids. A review. AB - The dinitrophenyl (DNP) derivatives of amino acids have found continual application in protein sequencing since Sanger used them for the first time for the sequencing of insulin. Dansyl derivatives of amino acids have been widely used in protein sequencing because of their fluorescent nature. The success of protein sequencing largely depends upon correct identification of such derivatives. The choice for the method of identification is related to cost, the availability of instrumentation and to the sensitivity needed for the analysis. Thin layer chromatography (TLC) is simple and has several advantages over other chromatographic methods. Therefore the literature after 1972 is reviewed for TLC analysis of dansyl- and DNP-amino acids, the two important amino acid derivatives required for identifying protein sequences. Additionally, the literature on the TLC resolution of enantiomeric mixtures of dansyl amino acids is reviewed. Application of various adsorbents, composition of solvent systems and other experimental conditions together with successful resolution data have been discussed. TLC provides a direct and inexpensive method for the resolution of enantiomers, and is fast becoming a sensitive instrumentalized quantitative analytical technique. PMID- 2559784 TI - Brain white-matter lesions and psychosis. PMID- 2559783 TI - Train-of-four as an index of neuromuscular block in cats: changes induced by atropine. AB - 1. The present study reevaluates the effect of atropine on the rate of recovery from tetanic fade caused by intraarterial administration of neostigmine or antinicotinic agents in cat anterior tibial muscle preparations submitted to a train-of-four (TOF) pattern of nerve stimulation. The study also compares the sensitivity of the TOF and tetanic responses as indices of residual nondepolarizing block. 2. Neostigmine, hexamethonium and d-tubocurarine all produced short-lived TOF fade. Both single and TOF twitches were increased by neostigmine and depressed by the antinicotinic agents. 3. Prior administration of atropine reduced the TOF fade induced by the antinicotinic drugs but potentiated that caused by anticholinesterase drugs. 4. These results indicate that TOF fade is not the most sensitive index for studying neuromuscular blockade when drugs other than neuromuscular blockers are also present. PMID- 2559785 TI - BRL 43694. PMID- 2559786 TI - Stimulation of limb cartilage differentiation by cyclic AMP is dependent on cell density. AB - Cyclic AMP (cAMP) has been implicated in the regulation of limb cartilage differentiation. This study represents an attempt to clarify potential mechanisms by which cAMP might regulate chondrogenesis. We have found that the ability of cAMP to stimulate limb cartilage differentiation in vitro is dependent on cell density. Dibutyryl cAMP (dbcAMP) elicits a striking increase in the accumulation of Alcian blue, pH 1.0-positive cartilage matrix, and a corresponding three- to fourfold increase in the accumulation of 35S-labeled glycosaminoglycans (GAG) by limb mesenchymal cells cultured in low serum medium at densities greater than confluence (i.e. micromass cultures established with 1-2 x 10(5) cells in 10 microliters of medium). Moreover, dbcAMP causes a striking (two- to fourfold) increase in the steady-state cytoplasmic levels of mRNAs for cartilage characteristic type II collagen and the core protein of cartilage-specific sulfated proteoglycan in these high density, supraconfluent cultures. In contrast, cAMP does not promote the chondrogenesis of limb mesenchymal cells cultured at subconfluent densities (i.e. cultures initiated with 2.5-5 x 10(4) cells in 10 microliters of medium). In these low density cultures, dbcAMP does not promote the formation of cartilage matrix, sulfated GAG accumulation or the accumulation of cartilage-specific mRNAs. These observations suggest that cAMP may exert its regulatory effect in part by facilitating cell-cell communication during the critical condensation phase of chondrogenesis. PMID- 2559787 TI - [Radiolocalization of human hepatoma with anti-human hepatoma monoclonal antibodies and its F(ab')2 in tumor-bearing nude mice]. AB - This paper reports the results and procedures for the radiolocalization of human hepatoma by murine MAb (HAb 18) and its F (ab')2 in animal model. 131I-labelled HAb 18 IgG and 125I-labelled its F(ab')2 were intravenously injected respectively to human hepatoma bearing nude mice. Clear tumor imaging manifested 48 hrs. after injection of 125I-labeled F(ab')2 and 120 hrs. after 131I-labeled intact MAb. The animals were killed 48, 72, 120, and or 168 hrs. after injection respectively. Bound radioactivity was counted in a variety of solid organs. Tumor: liver ratios for intact MAb or its F(ab')2 were greater than those for unrelated MAb at each time points, the differences being greatest after 168 hrs. (ratio intact MAb 5.15 +/- 0.64, ratio unrelated MAb 0.93 +/- 0.16) and after 48 hrs. (ratio F(ab')2 14.47 +/- 2.35, mean +/- sd). The amount of I-MAb bound was greater in the tumor than in any other solid organ. The clearance rate of F(ab')2 was faster then that of intact MAb. It was shown that HAb 18 MAb may be of value as an immunodiagnostic agent for hepatoma. PMID- 2559788 TI - [Localization of human lung cancer xenografts in nude mice using monoclonal antibody]. AB - Radiolabeled monoclonal antibodies to human lung cancer were injected into nude mice bearing xenografts; and the biodistribution and tumor localization were studied. Xenografts were able to concentrate radioiodinated McAb. Radioactivities of tumor which were 5.09, 5.79 and 7.57% dose/g respectively 3-5 days after injection were higher than those of blood or organ. Tumor/blood ratios were 1.82 2.36 whereas tumor/organ ratios were more than 2. The localization indices of two McAbs were 3.4-5.6 and less than 1.5 in normal organs. The xenografts were visualized after injection of radiolabeled McAbs. These results have shown that anti-human lung cancer McAbs can specifically localize on xenografts in nude mice and it is possible to use them clinically for imaging of lung cancer. PMID- 2559789 TI - Mullerian adenosarcoma of uterus--a case report. AB - A rare case of Mullerian adenosarcoma occurred in the uterine endometrium of a 40 year-old woman. Histologically, the tumor was composed of an admixture of benign appearing glands and a sarcomatous stroma. This neoplasm appears to have a relatively low malignant potential in contrast to the more aggressive form of malignant mixed Mullerian tumor. Our case was treated with hysterectomy and radiation. She was well after two and a half years. PMID- 2559790 TI - Effect of the superoxide dismutase inhibitor, diethyldithiocarbamate, on the cytotoxicity of mitomycin antibiotics. AB - Mitomycin C (MC) and its structural analogs porfiromycin (PM), BMY-25282 and BL 6783 are toxic to EMT6 cells under aerobic and hypoxic conditions. The mitomycin antibiotics are hypothesized to exert cytotoxicity under hypoxic conditions by cross-linking DNA following reductive activation, while aerobic cytotoxicity may involve DNA cross-linking by these agents and/or damage due to the generation of oxygen radicals. Previous findings (Pritsos and Sartorelli, 1986) indicated that the rank order of cytotoxicity for a series of mitomycins was the same as the rank order for the rate of oxygen consumption induced by these agents. As an additional approach to explore the role of oxygen radicals in the aerobic cytotoxicity of the four agents studied, EMT6 cells were treated with the mitomycins in the presence of the superoxide dismutase inhibitor diethyldithiocarbamate (DETC). DETC, which decreased superoxide dismutase activity in EMT6 cells, increased the cytotoxicity of BMY-25282 and BL-6783 by half an order of magnitude, but did not affect the toxicity of PM or MC to these cells. DNA cross-links, a proposed cytotoxic lesion induced by BMY-25282, however, were not detectably increased in EMT6 cells exposed to this agent in the presence of DETC in spite of the large increase in cytotoxicity under these treatment conditions. No single strand breaks were detected in cells exposed to either BMY-25282 or BMY-25282 plus DETC. The findings support the concept that oxygen radicals may have a role in the aerobic cytotoxicity of some of the mitomycin antibiotics, and that the lesions responsible for cytotoxicity produced by oxygen radicals may not reside entirely at the level of DNA. PMID- 2559791 TI - Synthesis of the 4-acetamido-4-deoxy analogue of N-acetylneuraminic acid and its behaviour towards CMP-sialate synthase. PMID- 2559792 TI - The side-chain conformations of N-acetyl-7-,8-,9-deoxy-, and -4,7-dideoxy neuraminic acid and their effect on the activation of CTP:N-acylneuraminic acid cytidylyltransferase. AB - The conformations of the deoxy side-chain analogs of N-acetylneuraminic acid and of N-acetyl-4-deoxyneuraminic acid have been studied by n.m.r. spectroscopy, which allowed us to distinguish between local minima conformations suggested by hard-sphere calculations. The conformations were correlated with the activity with CTP: N-acetylneuraminic acid cytidylyltransferase (CMPsialate synthase; EC 2.7.7.43). PMID- 2559793 TI - Pathogenesis and prevention of hepatocellular carcinoma. PMID- 2559794 TI - Hepatitis B virus and hepatocellular carcinoma--treatment of HBV carriers with Phyllanthus amarus. AB - Extracts of Phyllanthus amarus inhibit the DNA polymerase of HBV and related viruses. Woodchuck carriers of woodchuck hepatitis virus (WHV) were treated intraperitoneally with P. amarus extract. Three of four animals which had been recently infected lost the virus. Animals infected for about 3 months or more had a decrease in virus levels. Human carriers of HBV were treated orally for 1 month. About 60% of the carriers lost HBV, which did not return during the observation period. Fractions containing active principles are now being isolated and characterized. PMID- 2559795 TI - Does non-A, non-B hepatitis cause hepatocellular carcinoma? AB - The mechanism by which HBV infection leads to hepatocellular carcinoma is not as well defined as one would wish. While integration of viral DNA into host chromosomal DNA may be an important mechanism, especially in relatively "normal" livers, another mechanism more closely related to chronic cell death and regeneration resulting from chronic hepatitis is probably also important. Thus, hepadnaviral hepatocarcinogenesis may be multifaceted. Although it is not known whether the genome of HCV can integrate into host chromosomal DNA (it probably cannot), HCV can lead to chronic infection, chronic hepatitis, and cirrhosis in a significant proportion of patients, and there is growing epidemiologic evidence that such disease leads to HCC. There is less evidence that HDV is etiologically associated with HCC and the outcome of chronic HDV infections may be determined by the balance between the potentiating and inhibitory effects of HDV on the underlying HBV infection. PMID- 2559796 TI - Dietary risk factors for primary hepatocellular carcinoma. AB - Primary hepatocellular carcinoma (PHC) is among the most common forms of cancer, on a global basis. Incidence of the disease varies greatly in different areas of the world, suggesting involvement of environmental etiological factors, and much research has been devoted to their identification. Because many organic chemicals have been shown to have the capability of inducing PHC in animals, they have been extensively studied with respect to their possible significance as etiologic agents for PHC in man. Particular emphasis has been placed on aflatoxins because of their widespread occurrence as food contaminants and potency as liver carcinogens in a large number of experimental animals, including subhuman primates. Epidemiologic surveys have revealed a strong statistical correlation between aflatoxin ingestion and incidence of PHC in several areas of Africa and Asia. The IARC has recently determined the combined experimental and epidemiological information to be sufficient to designate aflatoxins as human carcinogens. The importance of hepatitis B virus (HBV) infection as a risk factor for PHC is extensively discussed by other authors in this journal issue. Prospective epidemiological studies have shown a high incidence of PHC among HBV carriers in HBV-endemic areas. Clinical studies have also shown that most PHC patients are carriers of the hepatitis B surface antigen and have chronic active hepatitis. Recently, HBV sequences have been found to be integrated into the liver cell genome in some, but not all, patients with chronic hepatitis or PHC. This evidence has identified HBV as a major etiological factor for PHC in certain populations, particularly in Taiwan and the People's Republic of China.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559798 TI - Nonhuman primate models for non-A, non-B hepatitis. AB - The three epidemiological forms of human non-A, non-B hepatitis have been studied by experimentally infecting non-human primates. The chimpanzee (Pan troglodytes) has been widely studied as a model for the epidemiological forms referred to as "blood-transmitted" and "coagulation-factor-transmitted" non-A, non-B hepatitis. Transmission of the "enteric" epidemiological form of non-A, non-B hepatitis to chimpanzees has been reported by one laboratory but remains to be confirmed. Two marmoset (tamarin) species, Saguinus mystax and S. labiatus, have been experimentally infected with the "blood-transmitted" form; S. mystax has been infected with the "enteric" form. A preliminary report suggesting susceptibility of marmosets to the "coagulation-factor-transmitted" form needs to be confirmed. The cynomolgus monkey (Macaca fascicularis) has been infected with the "enteric" form. Although the relationship between non-A, non-B hepatitis and hepatocellular carcinoma (HCC) is unclear, HCC has been reported in one chimpanzee, nearly 7 years after experimental infection with a non-A, non-B hepatitis virus. PMID- 2559797 TI - Induction of hepatocellular carcinoma in nonhuman primates by chemical carcinogens. AB - Several compounds were evaluated in nonhuman primates for their potential to induce neoplasms, especially hepatocellular carcinoma (HCC). The compounds can be classified into three groups: food contaminants, model rodent carcinogens, and nitrosamines. All three compounds in the food contaminants group, namely, aflatoxin B1, sterigmatocystin, and methylazoxymethanol acetate, induced HCC. None of the model rodent carcinogens tested consistently induced HCC in rhesus and cynomolgus monkeys. Three of four nitrosamines evaluated induced HCC in rhesus and cynomolgus monkeys. One nitrosamine, diethylnitrosamine, is a predictable and potent inducer of HCC and is useful for establishment of a nonhuman primate model for numerous oncologic studies. PMID- 2559799 TI - Hepadnaviruses and hepatocellular carcinoma (HCC). AB - Members of the hepadnavirus family share properties of virion structure, genome structure and replication, epidemiologic behavior, and pathogenic effects. Persistent infections with hepatitis B virus (HBV) in man, woodchuck hepatitis virus (WHV) in Marmota monax, ground squirrel hepatitis virus (GSHV) in Spermophilus beecheyi, and duck hepatitis B virus (DHBV) in domestic ducks of China are associated with development of hepatocellular carcinoma (HCC). Epidemiological evidence implicating hepadnavirus infection in HCC includes the observation that the geographic distributions of HBV infection and HCC are similar, that the incidence of HCC is much higher in hepadnavirus-infected than uninfected hosts, and that viral DNA sequences are integrated in the cellular DNA of most (e.g., 80 to 90%), but not all, hepadnavirus-associated HCC. Cirrhosis further increases the risk of HCC in HBV-infected humans. The precise role of hepadnaviruses in development of most HCC is unclear, although the finding of viral integrations within or near protooncogenes in a few cases suggests the possibility that these integrations may play a direct role in these HCC. However, in the great majority of HCC, viral integrations are in different cellular DNA sites in different HCC, integrations are not within domains of known protooncogenes, and integrations are not found in some 10 to 15% hepadnavirus associated HCC, suggesting that persisting viral sequences are not directly involved in the development of these HCC as viral sequences are for tumors caused by viruses with oncogenes or viruses that act by a "promoter-insertion" mechanism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559800 TI - Mitotic abnormalities leading to cancer predisposition and progression. AB - The development of human cancer is generally thought to entail a series of events that cause a progressively more malignant phenotype. Such a hypothesis predicts that tumor cells of the ultimate stage will carry each of the events, cells of the penultimate stage will carry each of the events less the last one, and so on. That is to say a dissection of the pathway from a normal cell to a fully malignant tumor may be viewed as the unraveling of a nested set of aberrations. In experiments designed to elucidate these events, we have compared genotypic combinations at genomic loci defined by restriction endonuclease recognition site variation in normal and tumor tissues from patients with various forms and stages of cancer. The first step, inherited predisposition, is best described for retinoblastoma in which a recessive mutation of a locus residing in the 13q14 region of the genome is unmasked by aberrant, but specific, mitotic chromosomal segregation. A similar mechanism involving the distal short arm of chromosome 17 is apparent in astrocytic tumors and the event is shared by cells in each malignancy stage. This is distinct from a loss of heterozygosity for loci on chromosome 10 which is restricted to the ultimate stage, glioblastoma multiforme. These results suggest a genetic approach to defining degrees of tumor progression and means for determining the genomic locations of genes involved in the pathway as a prelude to their molecular isolation and characterization. PMID- 2559801 TI - Multifactorial etiology and multifaceted prevention strategy of hepatocellular carcinoma. AB - In many Asian and African countries, hepatitis B virus (HBV) has been identified as a major causative agent of hepatocellular carcinoma (HCC). In these endemic areas, HBV infection also occurs early in life. An internationally collaborative study on universal immunization of newborns against HBV to prevent HCC was initiated in September 1983 and expanded later in the rural Qidong County of China, a prevalent area of both HBV infection and HCC. The 5-year follow-up data of the pilot phase of this controlled study have shown encouraging results. Over 98% of the newborns in local communities were vaccinated and over 97% of the vaccinees are being followed. The vaccination group with boosted 5-micrograms regimen showed significantly reduced HBsAg positive rate at 5 years, 2.5 vs. 12.5% of the nonvaccinated age-matched control. The anti-HBs level was also enhanced. The main study starting from the beginning of 1985 has been progressing according to schedule. Striking male preponderance and peak age shift to younger age of HCC in the high incidence area strongly points to the action of cofactors that may further increase the risk. Male preponderance was not observed in the animal models of HCC induced by hepadnaviruses. These phenomena were associated with increased exposure to aflatoxins as measured by urinary excretion of AFM1, as well as with a high prevalence of tobacco smoking and alcohol consumption among the Chinese males. The clarification of the etiological role of cofactors has important implications in the possible prevention of HCC among HBV carriers. PMID- 2559802 TI - HIV gene regulation and pathogenesis. AB - Human immunodeficiency virus (HIV) is unique among retroviruses in its genetic complexity. Its genome encodes a number of positive, differential, and negative regulatory genes, whose interplay appears to be directed at maintaining a steady, low-level virus expression. Since clinical progression and CD4 cell depletion in HIV-infected individuals appear to correlate with increase in virus expression, and continuous recruitment of new infected cells, the role for cofactors which enhance HIV production becomes significant in the pathogenesis of AIDS. Many environmental and cellular factors have been found to activate HIV. In particular, some viral agents may interact with HIV in contributing to pathogenesis. The leukemia viruses, HTLV-1 and HTLV-2, and several herpesviruses have been shown to stimulate gene expression from the HIV LTR. In addition, HIV tat gene can also activate a DNA virus (JC virus) which is associated with a neurological disease. Finally, immunosuppression by HIV is likely to reactivate latent herpesvirus infections, thus initiating a vicious cycle for further CD4 cell depletion. PMID- 2559803 TI - Circulating calcium modulates adrenaline induced cyclic adenosine monophosphate production. AB - Inotropic support of the failing myocardium may combine calcium with adrenaline in an attempt to augment the haemodynamic actions of each drug. We have previously shown, however, that calcium blunts adrenaline induced increases in blood pressure and cardiac output in animals and man. The mechanisms by which calcium may interfere with the haemodynamic actions of adrenaline are not well understood. Adrenaline is known to stimulate adenylate cyclase and increase levels of cellular cyclic adenosine monophosphate (cAMP), a crucial second messenger in cell regulation. We evaluated the effect of increased circulating calcium levels on adrenaline stimulated cAMP production in laboratory animals. Calcium infusion in rats nearly doubled the circulating ionised calcium concentration, from 1.27 (SEM 0.03) mM to 2.3(0.18) mM. Adrenaline infusion significantly increased plasma cAMP in saline infused control animals, from 26(6) pmol.ml-1 to 98(22) pmol.ml-1, whereas there was no increase in cAMP plasma levels in the calcium infused rats. The apparent inhibition of adenylate cyclase by calcium may participate in a negative feedback system which helps protect cells from harmful intracellular calcium overload. PMID- 2559804 TI - Dissociation between reperfusion induced arrhythmias and increases in ventricular alpha 1 receptor density in the anaesthetised rat. AB - Using anaesthetised rats we have assessed (1) whether the density of alpha 1 adrenergic receptors increases during coronary artery occlusion, (2) whether any change in density can be associated with the onset of reperfusion induced ventricular fibrillation, and (3) whether alpha 1 blockade with prazosin modifies the incidence of reperfusion induced ventricular fibrillation. The incidence of fibrillation upon reperfusion after 3, 5, 10, 20 and 30 min occlusion was 20, 75, 50, 16 and 10% (n = 10-12 in each group) respectively. alpha 1 Receptor density was measured using [3H]-prazosin in non-ischaemic and ischaemic tissue obtained after 0, 5 and 30 min ischaemia. Receptor density was not significantly altered at the time of maximum incidence of reperfusion induced ventricular fibrillation (5 min occlusion) but did significantly increase in both non-ischaemic and ischaemic tissue after 30 min occlusion, when the incidence of fibrillation upon reperfusion was very low (8%). At this time the values were 17.0(SEM 2.3) and 18.4(0.6)fmol.mg-1 protein in non-ischaemic and ischaemic zones as compared to 10.7(0.6) and 12.8(1.0)fmol.mg-1 protein in sham operated control animals (p less than 0.05 in both cases). Prazosin (0.1 or 1.0 mg.kg-1 body wt intravenously, 5 min prior to coronary occlusion) did not alter the incidence of ventricular fibrillation, ventricular tachycardia or total number of premature ventricular complexes upon reperfusion. We conclude that ischaemia induced changes in alpha 1 receptor density do not parallel changes in vulnerability to reperfusion induced arrhythmias.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559806 TI - Comparative effects of the opioids fentanyl and buprenorphine on ventricular vulnerability during acute coronary artery occlusion. AB - Fentanyl, a mu selective opioid agonist in wide clinical use, raises the ventricular fibrillation threshold in the normal canine myocardium. We have previously shown that this effect is amplified by haemorrhagic stress. In order to determine if mu receptor activation is antifibrillatory during acute myocardial ischaemia, we compared the effects of two mu selective agents, fentanyl and buprenorphine, in open chest chloralose anaesthetised dogs. Each drug was administered intravenously in two doses 1 h apart (fentanyl 30 micrograms.kg-1.dose; buprenorphine 0.3 mg.kg-1.dose). Ventricular fibrillation threshold was measured during right ventricular pacing using the single stimulus technique. The threshold was determined before and during a 10 min left anterior descending coronary artery occlusion. Prior to fentanyl administration, ventricular fibrillation threshold decreased from a control value of 19(SEM 2) mA to 12(1) mA during coronary artery occlusion. After the first dose of this drug an attenuation in the ischaemia induced fall in fibrillation threshold from 23(4) mA to 15(2) mA was observed. After the second dose of fentanyl the decline in fibrillation threshold was significantly blunted at 22(4) mA during control and 18(3) mA during occlusion, p less than 0.05 compared to no drug. In an additional series of experiments atropine sulphate abolished the antifibrillatory action of fentanyl, indicating that vagal efferent activation is responsible for the protective effect of the drug during acute myocardial ischaemia. This is in contrast with its mode of action during haemorrhage, when it enhances vagal afferent inhibition of sympathetic tone, and atropine pretreatment is without effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559805 TI - Density of beta adrenoceptors in rat heart and lymphocytes 48 hours and 7 days after acute myocardial infarction. AB - Down regulation of the beta adrenoceptor is thought to play an important role in the diminished response to catecholamines in heart failure. beta Adrenoceptor densities were measured on membrane homogenates of rat right ventricle and lymphocytes 48 h or 7 d after experimental myocardial infarction, and in rats exposed to a continuous infusion of isoprenaline (400 micrograms.kg-1.h1). The performance of the rat hearts was also evaluated 48 h post infarction in an isolated retrograde perfused heart preparation. In contrast to a 60% down regulation in right ventricle and a 20% down regulation in lymphocyte membranes after isoprenaline infusion, there was no change in right ventricle and lymphocyte beta adrenoceptor densities after myocardial infarction. Left ventricular contractile performance was significantly depressed 48 h after myocardial infarction. Mean basal left ventricular pressure decreased from 108(SEM 3) to 63(4) mm Hg while the maximal response to dobutamine was decreased from 204(4) to 105(12) mm Hg (n = 8). No correlation was found between the receptor densities of right ventricular and lymphocyte membranes. We conclude that diminished response to beta sympathomimetics after myocardial infarction cannot be attributed to a loss of surface beta adrenoceptors, and that the lymphocyte beta adrenoceptor does not provide an adequate system to monitor small receptor changes on the myocardium. PMID- 2559808 TI - Distribution and subtype determination of beta-receptors in the spinal cord of the adult rat. AB - 1. We determined the number of beta-receptors in the whole spinal cord of the adult rat and in the cervical, thoracal, and lumbal/sacral parts. 2. The undivided spinal cord contains 47 +/- 10 fmol/mg beta-receptors (KD = 2066 +/- 982 pmol/liter), and the cervical part of the spinal cord contains 53 +/- 8 fmol/mg protein (KD = 3224 +/- 1775 pmol/liter). The thoracal part shows 40 +/- 1 fmol/mg protein (KD = 3229 +/- 104 pmol/liter), and the lumbal/sacral spinal cord contains 48 +/- 8 fmol/mg protein (KD = 3610 +/- 1610 pmol/liter). 3. Competitive inhibition studies with l-practolol, dl-atenolol, and ICI 118,551 were performed and we calculated by a computer program in the whole spinal cord the following ratio of beta-receptor subtypes: 80 +/- 5% Beta 1-receptors and 20 +/- 5% beta 2 receptors. 4. The basal and (-)-isoproterenol- and NaF-stimulated activity of adenylate cyclase was highest in the cervical part of the spinal cord and equally distributed between the thoracal and the lumbal/sacral parts. 5. The whole synaptosomal protein of the cervical part of the spinal cord contained 132 +/- 20 fmol, the thoracal part 117 +/- 3 fmol, and the lumbal/sacral part 133 +/- 22 fmol. PMID- 2559807 TI - Serotonin-stimulated biochemical events in the procerebrum of Limax. AB - 1. The procerebrum (PC) of the terrestrial slug Limax maximus is of interest as a potential site of olfactory information processing (Gelperin et al., 1989). The neuromodulator serotonin is present in the procerebrum and can elicit action potentials from cultured procerebral neurons. We have investigated the effects of serotonin on second-messenger signaling systems and protein phosphorylation as a prelude to studies on long-term synaptic plasticity in the Limax procerebral lobe. 2. We found that several biochemical changes are triggered within 20 min of adding serotonin to the isolated procerebral lobe: adenylate cyclase is activated, protein phosphorylation and synthesis are modulated, and phosphatidylinositol-metabolism is stimulated. 3. Serotonin causes a rapid synthesis of cAMP, reaching a 20- to 30-fold increase within 1 min. Serotonin affects the rate of phosphorylation of several proteins, detected after a brief (20-min) incubation of the procerebral lobe in [32P]phosphate-containing medium. The level of synthesis of several proteins is altered by serotonin, as determined by alterations in [35S]methionine incorporation during a 20-min incubation. Serotonin also causes a slow accumulation of inositoltrisphosphate. 4. Our study shows that within a short time (less than 20 min) serotonin can influence several second-messenger signaling systems and the functional state and abundance of proteins in the procerebral lobe. These serotonin-stimulated events should have direct consequences for intercellular communication in the odor-processing network of the procerebral lobe. PMID- 2559810 TI - Glucose increases cytosolic calcium concentration and inositol lipid metabolism in HIT-T15 cells. AB - We have investigated the effects of glucose on cytosolic free calcium concentration in the insulin-secreting cell line HIT-T15. Addition of glucose (10 mM) caused a 20-75% increase in cytosolic [Ca2+] within 5 minutes compared to controls in the absence of glucose. A maximal increase in cytosolic [Ca2+] was obtained with 5 mM glucose. The magnitude of the response was markedly dependent upon the concentration of extracellular Ca2+, and the rise in cytosolic [Ca2+] was inhibited by verapamil. Cytosolic [Ca2+] was greatly increased by depolarization of the cells with KCl (50 mM), whereas carbamylcholine had no apparent effect. Glucose and KCl were also effective in stimulating insulin release from HIT cells, although carbamylcholine was again ineffective. The secretory response to glucose was also found to be directly related to the concentration of extracellular [Ca2+]. Glucose and KCl, but not carbamylcholine, were found to slightly enhance the production of [3H]-inositol trisphosphate in HIT cells pre-labelled with myo-[3H]-inositol, indicating a modest stimulation of inositol lipid hydrolysis. PMID- 2559809 TI - Does beta-adrenoceptor activation stimulate Ca2+ mobilization and inositol trisphosphate formation in parotid acinar cells? AB - The effects of the beta-adrenoceptor agonist, isoprenaline, on Ca2+ mobilization and inositol phosphate formation in parotid acinar cells were examined. Isoprenaline (2 microM) failed to increase cytosolic [Ca2+] in acinar cells, as measured by Fura-2 fluorescence, even in the presence of a phosphodiesterase inhibitor. Likewise, neither the 8-bromo nor the dibutyryl derivatives of cAMP (both at 2 mM concentration) increased [Ca2+]i. However, in confirmation of results previously published, a higher concentration of isoprenaline (200 microM) increased cytosolic [Ca2+]i of rat parotid acinar cells, from 104 +/- 4 nM to 151 +/- 18 nM. The increase in [Ca2+]i in response to isoprenaline, while transient in the absence of extracellular Ca2+, was sustained in Ca2(+)-containing medium. This isoprenaline-stimulated Ca2+ signal was more potently antagonized by phentolamine than by propranolol, suggesting that the higher concentration of isoprenaline activated alpha-adrenoceptors. Furthermore, the Ca2+ signal generated in response to the alpha-adrenoceptor agonist, phenylephrine, also was blocked by the same concentrations of propranolol necessary to block the effects of isoprenaline, suggesting that propranolol may block alpha-adrenoceptors under certain experimental conditions. The high concentration of (-)isoprenaline (200 microM) also increased inositol (1,4,5) trisphosphate and inositol (1,3,4) trisphosphate formation 45% within 30 s. Analogous to the increase in intracellular Ca2+, the formation of inositol phosphates stimulated by isoprenaline was more potently antagonized by the alpha-adrenoceptor antagonist, phentolamine, than by the beta-adrenoceptor antagonist, propranolol, again suggesting that isoprenaline interacts with alpha-adrenoceptors on parotid cells. Thus, the effects of isoprenaline on [Ca2+]i do not appear to be mediated by cAMP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559812 TI - Malignant Cystosarcoma phyllodes: report of two cases. AB - Two cases of malignant Cystosarcoma phyllodes seen in Jos University Teaching Hospital over a 9-year period are being reported. One occurred in a 16-year girl now dead and the other in a 52-year old woman probably dead. Both cases showed metastases, the axillary nodes being involved in the deceased, cervical nodes in the elderly patient, and both had pulmonary and presumably hepatic metastasis. These were the 2 cases seen out of a total of 696 surgical breast specimens reviewed thus constituting 0.3 percent. It is not certain if chemotherapy has been beneficial but one might need more cases well organised trials to arrive at definitive conclusions. PMID- 2559811 TI - Ca2+ and calmodulin-sensitive inositol trisphosphate kinase from bovine parathyroid. AB - A Ca2+ and calmodulin-activated inositol 1,4,5 trisphosphate kinase activity was detected in both soluble and membrane fractions from bovine parathyroid glands. Ca2+ activated the soluble enzyme in the concentration range 100 nM to 1 microM, which corresponds to the Ca2+ concentration range observed in the intact cell following maximal variation in extracellular Ca2+, the principal regulator of parathyroid hormone release. The Ca2+ sensitivity of the enzyme was absolutely dependent upon calmodulin. A similar activity was detected in the membranes but could be progressively removed by repeated washing at low ionic strength. This, together with data demonstrating binding of the enzyme to the hydrophobic matrix, Phenyl-Sepharose, suggests that the association of the enzyme with the membrane is likely to involve a significant hydrophobic component. The organic base, amiloride was identified as an inhibitor of the activity, the degree of inhibition being most marked in the presence of Ca2+ and calmodulin (K0.5 approx. 0.1 mM). The Ca2+ concentration dependence of the IP3 kinase suggests that inositol 1,3,4,5 tetrakisphosphate may be a messenger in the signal transduction pathway for the feedback inhibition of PTH secretion by extracellular Ca2+. PMID- 2559814 TI - Dynamic distribution of the Golgi marker thiamine pyrophosphatase is modulated by brefeldin A in rat hepatoma cells. AB - Cytochemical electron microscopy of cultured rat hepatoma cells (AH-130) demonstrated that thiamine pyrophosphatase (TPPase) activity was localized in the Golgi complex. When the cells were treated with brefeldin A (BFA, 2.5 micrograms/ml) for 10 min, the characteristic structure of the Golgi stack was no longer observed, and TPPase was cytochemically stained in the vesicular and tubular structures scattered in the cytoplasm. A longer exposure of the cells to the drug (20 min to 1 h) resulted in the distribution of the TPPase activity in the endoplasmic reticulum (ER) and nuclear envelope. Such an unusual distribution of the enzyme activity, however, was reversible even in the presence of BFA. At 2 h after the exposure, the TPPase activity disappeared from the ER and was concentrated again in the vesicular and tubular structures. The enzyme activity was finally localized in the Golgi complex which was reassembled by 4 h after the exposure. The reversible effect of BFA may be due to a possible metabolism of the drug into an inert form during the incubation. Taken together, these results indicate that BFA causes a rapid disassembly of the Golgi complex and redistribution of the marker enzyme TPPase into the ER including the nuclear envelope. The spontaneous reversibility of the drug effect also favors a dynamic recycling of the Golgi marker between the ER and the Golgi complex under the conditions used. PMID- 2559813 TI - 5'-Nucleotidase in rat liver lysosomal membranes is anchored via glycosyl phosphatidylinositol. AB - We have previously demonstrated that 5'-nucleotidase, known as a plasma membrane enzyme, is also distributed both in rat liver tritosomal membranes and contents (J. Biochem. 101, 1077-1085, 1987). When the lysosomal membranes isolated from rat livers were incubated with phosphatidylinositol-specific phospholipase C purified from B. thuringiensis, about 70% of 5'-nucleotidase activity was released from the membranes. Judging from the result by phase separation with Triton X-114, the enzyme solubilized by the phospholipase C digestion showed a hydrophilic nature such as that of the tritosomal contents. Immunoblot analysis showed that the molecular weight of 5'-nucleotidase released from the lysosomal membranes by the phospholipase C digestion was almost identical with that of the enzymes from the Tritosomal contents. The above results showed that the phosphatidylinositol-specific phospholipase C-like enzyme in the lysosomes may be responsible for the conversion of the lysosomal membrane-bound 5'-nucleotidase to the soluble form present in the lysosomal matrix. PMID- 2559815 TI - Isolation of an aggregation-less mutant of Dictyostelium discoideum with the expression of contact site A glycoprotein. AB - We isolated mutants defective in aggregation (aggregation-less) by mutagenizing the "double-bypass" mutant HG592 of Dictyostelium discoideum as the parental strain. One of the mutants expressed the contact site A glycoprotein with an apparent molecular weight of 80 X 10(3) on the cell surface in the normal developmental stage and retained EDTA-stable cell contact as well as EDTA sensitive cell contact. However, the mutant failed to aggregate on agar plates with bacteria. This mutant was designated HG700. We could not identify any differences between this mutant and the parental strain in levels of adenylate cyclase or extracellular phosphodiesterase activity, or in its chemotaxis toward cAMP. The mutant had greatly decreased the incorporation of [35S] sulfate into the particulate fractions of the cells starved for 6 h. This suggests that the modification by sulfation may crucially affect the mechanism of cell aggregation. PMID- 2559816 TI - [Multifocal chorioretinitis in spontaneous or induced immunodeficiencies]. AB - Multifocal simultaneous retinochoroditis are more and more frequent. They seem improved by immunodeficiencies. In 12 cases observed during only the 1987 year: 5 were spontaneous and 7 in relation with a corticosteroid treatment associated or not with other immunosuppressors. The vitreous products analysis lead in 10 cases to find 7 toxoplasmosis in 2 AIDS and 3 CMV with 2 positive Elisa test. The immunodeficiencies prevention spontaneous but also in relation with uncontrolled treatments appear as a very important necessity and urgency. PMID- 2559817 TI - [Optic neuropathy in Purtilo's syndrome]. PMID- 2559818 TI - Direct extraction and assay of collagenase from human osteoarthritic articular cartilage. AB - A method has been developed for the direct extraction of collagenase from small quantities (5 mg) of human osteoarthritic articular cartilage. The enzyme, which was not detected in normal cartilage, was entirely in a latent form and demonstrated typical properties of mammalian collagenase after activation by trypsin. PMID- 2559819 TI - Genetic analysis of familial amyloidotic polyneuropathy, an autosomal dominant disease. AB - Familial amyloidotic polyneuropathy (FAP) is an autosomal dominant genetic disease, and the major component of the amyloid fibrils from FAP patients was shown to be variants of transthyretin (TTR) with single amino acid substitutions. The nucleotide sequence analysis of the TTR cDNA and its corresponding gene enabled us to detect the base substitutions responsible for the variant TTR by conventional Southern blotting or polymerase chain reaction (PCR) method and allowed us to screen a number of FAP families in Japan. Among seven TTR variants related to FAP, all the FAP patients tested in Japan had the particular 30Val--- Met mutation. Haplotype analysis revealed that the Val----Met mutation has recurred frequently in the population to generate the FAP families of independent origins. Although the primary cause of FAP has become clear, extensive screening of FAP families revealed that there existed late onset cases and also patients with atypical symptoms in FAP families with the Val----Met mutation, suggesting that the expression of FAP is a complex process and affected by some (unknown) factors other than TTR. PMID- 2559820 TI - Phosphoinositide metabolism and oncogenes. AB - The hydrolysis of phosphatidylinositol 4,5-bisphosphate plays important roles in growth factor- or oncogene-induced cell proliferation. However, it is still unknown whether the hydrolysis of phosphatidylinositol 4,5-bisphosphate is an essential step for cell growth. To solve this problem, we developed a monoclonal antibody against the lipid. Injection of the antibody into ras-transformed cells caused reversible and dose-dependent decrease in DNA synthesis and reverted the cell morphology to that of the untransformed cells. The antibody also inhibited the proliferation of erbB- and src-transformed cells but not the proliferation of untransformed or myc-transformed cells. PMID- 2559821 TI - The bioactivity of immunoreactive adrenocorticotrophin in human blood is dependent on the secretory state of the pituitary gland. AB - A highly sensitive bioassay, using preincubated purified isolated rat adrenal cells, has been developed for measuring plasma ACTH. This bioassay enables detection of ACTH in plasma of about 0.9 pmol/l. Bioactive ACTH (B-ACTH) plasma levels were determined during insulin-induced hypoglycaemia in 12 female subjects and the values were compared with immunoreactive ACTH (I-ACTH) levels. The mean (+/- SD) basal B-ACTH level amounted to 1.7 +/- 0.8 pmol/l, the mean I-ACTH to 2.9 +/- 1.4 pmol/l. The highest mean B-ACTH plasma value was found 30 min after insulin injection: 14.7 +/- 15.7 pmol/l (I-ACTH: 12.8 +/- 9.9 pmol/l). By 90 min the B-ACTH level had returned to baseline (1.6 +/- 0.8 pmol/l), whereas the I ACTH level was still significantly higher (5.4 +/- 2.7 pmol/l) than at time zero. Remarkably, the B-ACTH to I-ACTH ratio (B/I ratio) showed a biphasic profile during the insulin tolerance test, the ratio increasing from 0.60 +/- 0.77 at time zero to 1.08 +/- 0.35 at the ACTH peak, and decreasing after that to a lower value of 0.33 +/- 0.11 at 90 min. From these results it is concluded: (1) in the morning hours a considerable amount of circulating I-ACTH has no steroidogenic activity; (2) the B/I ratio temporarily increases immediately after insulin injection but gradually decreases afterwards to values half the baseline level at 90 min. Whereas this decrease at 90 min can be explained by differences in disappearance rates, the increase of B-ACTH relative to I-ACTH at 30 min indicates that estimations of immunoreactive ACTH reflect the biological activity of newly released ACTH with greater precision than at steady state level. Thus, the B/I-ACTH ratio can be used as a tool for measuring the state of the pituitary releasing activity. PMID- 2559822 TI - Stimulation of cyclic AMP production in FRTL-5 thyroid cells by crude immunoglobulin fractions of serum from pregnant women. AB - In an assay for thyroid-stimulating antibodies, in which FRTL-5 thyroid cells were incubated with crude immunoglobulin (Ig) fractions precipitated from serum with 15% polyethylene glycol, significant increase in cAMP production was elicited by the samples from 25 (35.7%) out of 70 pregnant women. The highest value was 529.5%. There was a close correlation between thyroid stimulating activities and serum hCG concentrations (r = 0.708, P less than 0.001). When 125I hCG was added to serum from pregnant women, about 20% of the radioactivity was incorporated into the Ig fractions. hCG preparations within a range of concentrations of 30-300 IU/m elicited 2.3-16.5 times increase in cAMP in a dose dependent manner. Nine pregnant women with serum TSH concentrations less than the lower limit of the normal range (less than 0.25 mU/l) displayed significantly higher values for both thyroid stimulating activities and serum hCG concentrations (P less than 0.001, respectively) compared with those who had normal TSH levels in serum. These data suggest that hCG or its variant may stimulate the thyroid sufficiently to suppress secretion of TSH from the pituitary in some pregnant women. PMID- 2559823 TI - Megavoltage pituitary irradiation in the management of Cushing's disease and Nelson's syndrome: long-term follow-up. AB - We report the long-term follow-up of the clinical and biochemical effects of megavoltage pituitary irradiation (radiotherapy; RT), administered as primary or secondary therapy, for pituitary Cushing's disease and Nelson's syndrome in 52 patients. Irradiation was administered, from a 4-15 MeV linear accelerator, via a three-field technique (two lateral, one frontal), to a total dose of 4500 cGy (rad) in 25 fractions over 35 days. Twenty-one patients received RT as primary ablative therapy for Cushing's disease and were under follow-up 5.8 to 15.5 years later (median 9.5 years). All were initially treated with metyrapone to induce normal mean plasma cortisol levels, and all achieved clinical remission on this therapy. At latest follow-up, 12 (57%) are off all therapy, in clinical remission, with a normal mean cortisol through the day; however, only two show completely normal plasma cortisol responses to dynamic testing; four remain on medical therapy with metyrapone or op'DDD and all have required a steady dose reduction accompanied by falling plasma ACTH levels; five have required alternative therapy with bilateral adrenalectomy and/or transsphenoidal hypophysectomy. Fifteen patients received RT for Nelson's syndrome, developing after bilateral adrenalectomy, and have been followed up for 1.5 to 17.3 years (median 9.6 years). Fourteen patients showed progressive depigmentation, shrinkage of the pituitary adenoma and fall in plasma ACTH levels to 1-72% (median 16%) of the pre-RT basal value. In the remaining patient an initial fall in plasma ACTH was followed by tumour enlargement at 6 years, leading to death at 11 years after RT. Of the remaining patients, results are assessed in nine who received RT after unsuccessful transsphenoidal surgery, three after transfrontal surgery for aggressive macroadenomas, and four prophylactically after bilateral adrenalectomy. Radiotherapy remains a valuable second-line therapy for Cushing's disease and its complications. PMID- 2559824 TI - Radiation-induced hypopituitarism is dose-dependent. AB - Radiation-induced hypopituitarism has been studied prospectively for up to 12 years in 251 adult patients treated for pituitary disease with external radiotherapy, ranging in dose from 20 Gy in eight fractions over 11 days to 45 Gy in 15 fractions over 21 days. Ten further patients were studied 2-4 years after whole-body irradiation for haematological malignancies using 12 Gy in six fractions over 3 days and seven patients were studied 3-11 years after whole brain radiotherapy for a primary brain tumour (30 Gy, eight fractions, 11 days). Five years after treatment, patients who received 20 Gy had an incidence of TSH deficiency of 9% and in patients treated with 35-37 Gy, 40 Gy and 42-45 Gy, the incidence of TSH deficiency (22, 35 and 52% respectively) increased significantly (P less than 0.001) with increasing dose. A similar relationship was observed for both ACTH and gonadotrophin deficiencies when the 20 Gy group was compared to patients treated with 35-45 Gy (P less than 0.01 and P less than 0.05 respectively). Growth hormone deficiency was universal by 5 years over the dose range 35-45 Gy. In seven patients who were treated with 30 Gy in eight fractions over 11 days, deficiencies were observed at a similar frequency to the 40 Gy group (15 fractions, 21 days). No evidence of pituitary dysfunction was detected in the ten patients who received 12 Gy (six fractions, 3 days). Both total radiation dose and fractionation schedule may determine the incidence of pituitary hormone deficiencies. The dose below which deficiencies do not occur is probably irrelevant to therapeutic irradiation of pituitary and other intracranial neoplasms. PMID- 2559825 TI - Paraneoplastic opsoclonus: a neuropathologic study of two cases. AB - The association of opsoclonus and malignant neoplasia is infrequent. The clinical and neuropathological data of two patients in whom opsoclonus and ataxia developed 7 and 11 months before the detection of a bronchial carcinoma are reported. Loss of Purkinje cells, edema of dentate nucleus and peridental demyelination were the most important neuropathological findings; neither carcinomatous metastases nor inflammatory signs were found in the brain. From the review of the pathological reports of paraneoplastic opsoclonus, the following conclusions can be drawn: the changes in the cerebellum are produced by the paraneoplastic cerebellar degeneration and are unrelated to the origin of opsoclonus, which has other anatomic substrates; paraneoplastic opsoclonus is a "remote effect" of cancer with an inflammatory basis, for which neurotoxic and immunological mechanisms have been hypothesized. PMID- 2559826 TI - Silica prevents the induction of diabetes with complete Freund's adjuvant and low dose streptozotocin in rats. AB - Three weekly intraperitoneal injections of complete Freund's adjuvant (CFA) and, one day later, 25 mg/kg of STZ have been reported to cause a gradual onset of autoimmune diabetes. We have previously reported that the onset of diabetes in this model occurs too rapidly to be due to a specific immune reaction and that other compounds that activate macrophages also potentiate low-dose STZ in rats. We have proposed that mediators released from activated macrophages may enhance the cytotoxic effect of STZ. In the present study, we have observed that intraperitoneal injections of silica, a macrophage toxin, markedly decrease the incidence and severity of diabetes induced with CFA and low-dose STZ. PMID- 2559827 TI - Phosphoinositide-specific phospholipase C of a cloned mast cell line, mastocytoma P-815: purification and some characterization. AB - 1. Multiple forms of phosphoinositide-phospholipase C (PLC) were isolated from mastocytoma; two cytosolic forms (cPLC-I, Mr 150,000; cPLC-II, Mr 110,000) and two membrane-associated forms (mPLC-I, Mr 85,000; mPLC-II, Mr 85,000). 2. Four PLC forms differently behaved in substrate specificity and effect of GTP-binding proteins. PMID- 2559828 TI - Effect of oral contraceptives on adrenocorticotropin and growth hormone secretion following CRH and GHRH administration. AB - Previous investigations have demonstrated that basal ACTH plasma levels are reduced and that GH levels are either increased or unchanged in women taking oral contraceptives. The purpose of the present investigation was to determine the secretion of ACTH and GH following an intravenous infusion of human corticotropin releasing hormone (CRH) and human growth hormone releasing hormone (GHRH) in control women (N = 8) and in women taking a triphasic oral contraceptive (N = 9). The studies were initiated between 7:00 and 9:00 a.m. and all women were fasting. An intravenous catheter attached to a 3-way stopcock was used for blood sampling and to inject a bolus of CRH and GHRH (1 microgram/kg). Plasma samples were frozen immediately and stored at -70 degrees C until assayed for content of ACTH and GH by radioimmunoassay. The plasma levels of ACTH and GH increased following infusion of CRH and GHRH in all women. The mean plasma levels of growth hormone were not statistically different in oral contraceptive users compared to normal women. In contrast, ACTH plasma levels in oral contraceptive users were reduced approximately 25% overall, and significantly lower (p less than 0.04) at 120 minutes following the CRH infusion compared to controls. In conclusion, the GHRH stimulated GH release was similar in normal women and oral contraceptive users. CRH-stimulated ACTH release was modestly reduced in oral contraceptive users compared to normal women suggesting that estrogens and progestogens may have a suppressive effect on the release of ACTH by the pituitary. PMID- 2559829 TI - Transducin: a signaling switch regulated by guanine nucleotides. PMID- 2559830 TI - Coordinate interactions of cyclic nucleotide and phospholipid metabolizing pathways in calcium-dependent cellular processes. AB - It is hoped that his review enables the reader to appreciate the complexities implicit in the interactions among Ca2+, cyclic nucleotides, and phospholipid metabolizing pathways in cell signal transduction. The interactions are varied and intricate, often involving several levels of cell amplification mechanisms. Upsetting the balance of fatty acids in membrane phospholipids can have detrimental effects on adenylate cyclase. Thus, n - 3 fatty acid enrichment of phospholipids suppresses adenylate cyclase activity. The effects of significant alterations in dietary fatty acids, such as might occur with the current vogue for n - 3 eicosapentaenoic acid and docosahexaenoic acid (fish oil) dietary enrichment regimens, will need to be assessed more fully with regard to stimulus induced changes in cyclic nucleotide production in various tissues. Since the n - 3 fatty acids have not been demonstrated to affect guanylate cyclase activity, dietary changes in certain of these fatty acids would not be expected to contribute to changes in cGMP generation as much as in cAMP production. Moreover, the ingestion of large quantities of these n - 3 fatty acids can alter the profile of cyclooxygenase and lipoxygenase products produced in cells. According to the paradigm developed in this article, changes in the metabolism of fatty acids are amplified by alterations in cyclic nucleotide production and phospholipase activities, with the eventual physiological impact predicated on the tissue type and the specific stimulus response. There appears to be a rather clear distinction between the regulatory properties of eicosanoids regarding adenylate and guanylate cyclase activities. Whereas prostaglandins often stimulate adenylate cyclase activity, they have little effect on guanylate cyclase activity. On the other hand, the HETE compounds seem to play an important role in guanylate cyclase regulation in certain cells. Moreover, arachidonic acid affects adenylate cyclase activity without prior peroxidation, whereas endoperoxides and hydroperoxides are more effective than arachidonic acid with regard to guanylate cyclase stimulation. However, in the intact cell there is a strong implication that the dual stimulation of guanylate cyclase by Ca2+ and fatty acid evokes optimal enzyme activity. An advantage of multidimensional response mechanisms in cells includes the ability to recognize different stimuli and to respond with specific, coordinated responses modulated in their intensity and/or duration by messenger interaction. Few cell types respond to receptor stimulation in an all-or-none fashion, and the "milieu interior" depends on specific, graded responses to the autonomic nervous system and endocrine stimuli.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2559831 TI - [Intraoperative ultrasound of the pancreas in hyperinsulinism]. AB - In four cases of hyperinsulinism proven clinically and by blood chemistry the value of intraoperative sonography of the pancreas is emphasized. An insulinoma could be localized preoperatively in two patients. In one of these two cases intraoperative sonography detected additional insulinomas. In the remaining two patients the negative preoperative examinations and surgical palpation was confirmed by using this method: diffuse islet cell hyperplasia was the final diagnosis in both cases. PMID- 2559832 TI - [Yuanhuacin film for menstruation induction and termination of early pregnancy: analysis of 382]. AB - Yuanhuacin Film was used in 382 cases, 26 cases for menstrual induction (successful rate 100%) and 356 for termination of early pregnancy. Among the latter group 336 cases resulted in complete abortion (94.4%), 12 cases in incomplete abortion (3.4%) and 8 cases in failure (2.2%). The embryonic sacs were discharged within 34.66 +/- 33.73 (means +/- s) hours. The amount of vaginal bleeding was less than or equal to that of menstruation, which lasted for 8.18 + 9.14 (means +/- s) days with mild abdominal pains. In the complete abortion group 237 cases (70.5%) were followed up and 236 cases were found to restore their menses within 40 days (99.6%), and 157 cases have decreased amount or the same amount of menstrual flow (66.2%), while in 80 cases the amount was increased (33.8%). PMID- 2559833 TI - [Ultrastructural and histochemical studies of late infantile metachromatic leukodystrophy]. AB - Ultrastructure of cellular inclusions in the cerebral cortex and in renal tubule epithelial cells in a case of late infantile metachromatic leukodystrophy were investigated. Four patterns of shape were illustrated as multi-lamellated structure, zebra bodies, dense bodies and vacuoles with the two latter being more predominant in the renal tubule cells. Histochemical studies of the cellular deposits with toluidine blue and cresyl violet all revealed metachromatic in character. Significance of these findings and their pathogenesis were discussed. PMID- 2559834 TI - [Carcinomatous neuropathy: clinical and pathologic findings of sural nerve biopsy in 7 cases]. AB - Sural nerve biopsy was done 7 cases of cancer patients associated with peripheral neuropathy. There were 3 cases of lung carcinoma and one each of pancreas adenoma, seminoma, sigmoid carcinoma and chondrosarcoma of the femur. The neurological features manifested themselves with sensory pattern of neuropathy associated with ataxia in one case, sensorimotor neuropathy in 3 cases, and idiopathic polyneuropathy, peripheral neuropathy with proximal myopathy and neuropathy with paraneoplastic cerebellar syndrome each in one case, 6 patients showed neuropathy before malignancy was discovered and only one patient had neuropathy after the onset of carcinoma. Sural nerve biopsy studied in all the 7 patients with light and electron microscope revealed no infiltration of carcinomatous cells in the sural nerve fascicles. There was severe loss of myelinated fibers and severely axonal degeneration in one patient. Another patient showed segmental demyelination (5.03 x 10(3)/mm2). There was evidence of both axonal degeneration and demyelination associated with moderate reduction in the number of the myelinated fiber density ranging from 1.02 to 4.35 x 10(3)/mm2. In 6 cases, mononuclear cells were seen in nerve fascicles under the electron microscope. The characteristic pathological findings, their relation with the duration and onset of the cancer and some ideas regarding the pathogenesis are discussed. PMID- 2559835 TI - [Chemosensitivity of MGc 80-3 human gastric adenocarcinoma cells and its clinical significance]. AB - The survival curves of MGc80-3 cells exposed for 24 hr to various concentrations of 10 anticancer drugs were obtained by means of colony-forming assay. Comparison of drug doses required to reduce cell survival by 90% (ID90) with the clinically achievable peak plasma drug concentration (PPC) showed that MMC, ADM, and 5-Fu had strong cytotoxicity (PPC/ID90 = 18-36), whilst the other drugs were less effective (PPC/ID90 less than or equal to 11). The fact that MMC, ADM, and 5-Fu are most effective in the treatment of gastric cancer suggests that the cell line MGc 80-3 may still retain its original drug sensitivity which is consistent with that noted in clinical gastric cancer. This cell line could be useful in screening for new compounds with activity against this disease. PMID- 2559836 TI - [Surgical treatment of carcinoma of gastric cardia in aged patients--analysis of 101 cases]. AB - 101 aged patients with cardiac carcinoma resected by upper midline abdominal approach and left costal cartilage amputation were reported. The merits of the operation were: better exposure; good tolerance of patients with embarrassed heart and lung functions; un-influenced functions of the diaphragm and intrathoracic vital organs because neither the diaphragm was incised nor the thorax was opened and blood loss was less than 200 ml during the operation. The authors believe that this method is suitable for the aged patients and patients with deficient cardiopulmonary functions. Long-term follow up results are needed for critical evaluation. PMID- 2559837 TI - [Roentgeno-pathologic studies of peripheral pulmonary carcinoma]. AB - 122 patients with solitary pulmonary nodular lesions less than 3 cm in diameter admitted to our hospital from 1974 to 1985 are analysed. All lesions were negative by sputum exfoliative cytology and fiberoptic bronchoscopy. As lung cancer could not be excluded, exploratory thoracotomy was performed. They were diagnosed as primary peripheral pulmonary carcinoma by pathology. Of the 122 patients, 45 had adenocarcinoma, 42 squamous-cell carcinoma, 19 bronchiolo alveolar carcinoma, 9 small cell carcinoma, 4 large cell carcinoma and 3 mixed type carcinoma. Pathologic specimen was compared with X-ray chest films to study the pathologic basis of radiologic manifestations. As there were different histologic types in solitary peripheral pulmonary carcinoma, their pathologic processes were different. Therefore, the radiologic features were also different. Different points of the radiologic manifestations might be helpful to differentiate different histologic types of pulmonary carcinoma and increase the diagnostic accuracy for early pulmonary carcinoma. PMID- 2559838 TI - [Diagnosis and treatment of insulinomas--analysis of 6 cases]. AB - Six patients with insulinoma treated from 1977-1986 are analysed. All patients had symptoms of neuro-glucopenia, hypoglycemic coma and sympathetic irritation. The fasting blood glucose level was below 30 mg%. The fasting plasma IRI was 17.3 19.84 mu u/ml and the IRI/glucose ratio was greater than 0.3 in four patients. The symptoms disappeared after the tumor was removed in three patients. One patient was complicated by pancreatic fistula after tumor resection, but was cured by drainage. One patient who had multi-insulinomas undiagnosed preoperatively had Whipple's triad after operation. One was treated by other methods because of his cardiovascular disorders. The percutaneous transhepatic portal catheterization segment IRI assay is a reliable method, especially when combined with abdominal selective arteriography, for the pre-operative diagnosis of insulinoma. The IRI/glucose ratio is more reliable than either glucose or IRI alone. PMID- 2559839 TI - Prevention of experimental calcinosis by salmon calcitonin in rats. AB - The effects of salmon calcitonin on calcium overload and its deposition in the rat aorta was studied. Calcitonin administered 4 days to rats i.p. in doses of 10 IU/kg body weight blocked the development of calcinosis induced by a single dose of vitamin D3 (300,000 IU/kg body weight, orally). This was demonstrated biochemically, histochemically and by electron microscopic methods. PMID- 2559842 TI - Reports of two cases of selective adrenocorticotropin (ACTH) and growth hormone (GH) deficiency: differential diagnosis from cases with isolated ACTH deficiency associated with transient GH insufficiency. AB - An acquired partial pituitary insufficiency with selective ACTH and GH deficiency was demonstrated in two men aged 47 and 54, for which the clinical course over many years corresponds to Addison's disease. In one of the 2 cases, antibodies to anterior pituitary cell membrane, assayed by an immunofluorescence method with GH3 cells (rat GH and prolactin secreting cell) and AtT-20 cells (mouse ACTH secreting cell) as antigens, were positive. We also present a 55-year-old man with isolated ACTH deficiency associated with transient GH deficiency. In this case, hydrocortisone replacement corrected his subnormal, pre-therapy GH response to insulin tolerance and glucagon propranolol tests, although there was no response of serum GH to L-dops and arginine stimulation test before therapy. Selective ACTH and GH deficiency are very rare and the finding of transient GH insufficiency in a patient with isolated ACTH deficiency suggests that repeated testing while on hydrocortisone replacement therapy is of great diagnostic importance in order to distinguish between selective ACTH and GH deficiency and isolated ACTH deficiency accompanied by transient GH insufficiency. PMID- 2559841 TI - Glucagon synthesis with mRNA preparation of a glucagon-producing tumor (IT-1). AB - Nucleic acids were extracted from the tumor (IT-1) and purified to give poly (A) containing RNA, which was subjected to protein synthesis in vitro with a wheat germ extract. Gel-filtration (Bio Gel P-30) profiles of the translated product showed the presence of glucagon-like substance, and the results of treatment of fractions with glucagon antibodies (30K or K4023) showed the possibility that translated products contained true-glucagon. This confirms glucagon synthesis in IT-1. The molecular weight of the translated glucagon was estimated to be 3,000 from the K-value. The time courses of the glucagon synthesis were examined in cultured tumor cells (ITC-1) using 3H-leucine as a tracer. A large molecular weight protein was already detected after pulse labeling for 1 h. The amount of labeled glucagon in the cells was shown to be maximum at 1 h. True-glucagon was converted at 3 h to smaller molecular weight peptides which reacted with the C terminal antibody of glucagon. In vitro protein synthesis, peptides with molecular weights of around 10,000 were major products in 15-30 min. PMID- 2559843 TI - Chemical substance transport in soils and its effect on groundwater quality. AB - The problems of chemical substance applications in different spheres of industry and agriculture and their effects on groundwater quality and human health are described. Sources of groundwater contamination from industrial and municipal wastes, agricultural pollutants, etc., are listed. The experience in the application of chemical fertilizers and pesticides in the USSR is described. A brief estimation of groundwater salinity is given for various regions of the USSR where irrigation is practiced, as well as the experience in environmental protection. Special attention is given to methods of simulating water seepage and chemical substance transport in soils. Boundary problems for free-surface seepage and dissolved solids transport in porous media are stated, and methods of solution are described in the example of the hydrodynamic theory of seepage and dispersion. Some results of calculations with this method are presented. The influence of groundwater quality on the morbidity of the population is given and the main diseases and associated medical problems are listed. PMID- 2559840 TI - Evaluation of hypothalamic-pituitary function in a combination of tests with four hypothalamic releasing hormones and L-dopa in normal subjects and in patients with hypothalamic and/or pituitary disorders. AB - Hypothalamic-pituitary function was evaluated in a combination of tests with four hypothalamic releasing hormones (4RHs) and L-dopa in normal subjects and in patients with hypothalamic and/or pituitary disorders. Plasma concentrations of anterior pituitary hormones (GH, ACTH, TSH, PRL, LH and FSH) were measured before and after simultaneous iv administration of GHRH, CRH, TRH and LHRH. In addition, changes in the plasma levels of GHRH and GH were investigated before and after oral administration of L-dopa. Normal subjects showed appreciable responses to both tests. In five patients with hypothalamic disorders, the response of plasma anterior pituitary hormones varied, but plasma GHRH and GH did not respond to L dopa. Patients with idiopathic and postpartum hypopituitarism showed low response to 4RHs or none at all, but L-dopa evoked a normal GHRH response in 2 of the 4 cases having no GH response. In the patients with hypopituitarism due to resection of a pituitary tumor, the response of anterior pituitary hormones to 4RHs was low, and L-dopa administration induced a normal GHRH and low GH response in 5 out of the 7 cases. After 4RHs administration, the patients with ACTH deficiency syndrome showed different patterns of impaired ACTH secretion, and isolated, combined or limited ACTH reserve. Seven patients with anorexia nervosa showed exaggerated GH, delayed TSH and FSH, low ACTH and LH, that is, normal PRL response to 4RHs, but no response of plasma GHRH or GH to L-dopa, suggesting the presence of hypothalamic dysfunction. These results indicate that the combination of the 4RHs test and L-dopa test is a simple and useful means for evaluating hypothalamic-pituitary function by measuring the response of plasma GHRH and six anterior pituitary hormones in the patients with endocrine disorders. PMID- 2559844 TI - Impact of diffuse nitrate pollution sources on groundwater quality--some examples from Czechoslovakia. AB - In several regions of Czechoslovakia with intensive agricultural production, the correlation between the amount of nitrogen fertilizer applied and the nitrate content in groundwater has been recognized. Nitrate pollution of groundwater is considered to be the most serious source of nonpoint pollution in Czechoslovakia. A program of research into the effects of farming activities on groundwater quality in Czechoslovakia is under way on experimental fields (20 to 30 hectares) and, simultaneously, in regions in which shallow, vulnerable aquifers occur. The importance of the soil organic matter's stability for maintaining the groundwater quality is emphasized. Research based on nitrogen and organic carbon balance has shown that the restoration of a soil-groundwater system is a complicated process that usually requires changes in the extent and intensity of agricultural activities and consistent attention to the effects produced by natural conditions. Regional investigation of the impact of farming on shallow aquifers in the fluvial deposits of the Elbe River in Bohemia has proved the hydrochemical instability and vertical hydrochemical heterogeneity of these aquifers. The WASTEN deterministic model was used for modeling the transport and transformation of various types of inorganic fertilizers. The input data is based on laboratory and field measurements. Special topics are the verification of model calculations and the time and spatial variability of input data with respect to the unsaturated zone. The research results are being used for making regional and national agro-groundwater managerial schemes more precise, as well as for decision-making. PMID- 2559845 TI - 9-(2-Phosphonylmethoxyethyl)-2,6-diaminopurine (PMEDAP): a novel agent with anti human immunodeficiency virus activity in vitro and potent anti-Moloney murine sarcoma virus activity in vivo. AB - 9-(2-phosphonylmethoxyethyl)-2,6-diaminopurine (PMEDAP) is a potent inhibitor of the replication of human immunodeficiency virus (HIV) in human T-lymphocyte MT-4 cells (50% effective dose: 2 microM). PMEDAP strongly inhibited Moloney murine sarcoma virus (MSV)-induced transformation of murine C3H/3T3 embryo fibroblasts and caused a dose-dependent suppression of tumor formation and mortality in newborn mice inoculated with MSV. Even at a dose as low as 0.25 mg/kg/day, PMEDAP effected a significant delay in tumor appearance and an enhancement of the survival rate of tumor-bearing mice. PMEDAP proved fivefold more efficacious as an anti-MSV agent than 9-(2-phosphonylmethoxyethyl)-adenine (PMEA), which has been previously shown to exhibit strong antiretroviral efficacy in vivo. However, PMEDAP was also more toxic, so that its therapeutic index was equivalent to that of PMEA. PMID- 2559846 TI - Susceptibility of anaerobic bacteria to the new streptogramin RP 59500 in vitro. AB - The activity of the new streptogramin RP 59500 in vitro was determined against 380 strains of anaerobic bacteria by an agar dilution method and compared with that of pyostacine, piperacillin, cefoxitin, imipenem, clindamycin, metronidazole and chloramphenicol. RP 59500 and imipenem were the most active agents tested. On the basis of these results, RP 59500 appears to be a promising antimicrobial agent for treatment of anaerobic infections, further clinical investigations being warranted. PMID- 2559847 TI - Value of new quinolones in the treatment and prophylaxis of infectious diseases: introductory remarks. PMID- 2559850 TI - Coronary vasodilation induced by intracoronary enalaprilat: an argument for the role of a local renin-angiotensin system in patients with dilated cardiomyopathy. AB - Although indicated by several experimental studies, the presence of a renin angiotensin system has not been demonstrated in the human heart. The influence of a local renin-angiotensin system on the coronary vessels may be difficult to establish after oral or intravenous administration of an angiotensin converting enzyme inhibitor, since coronary blood flow depends greatly on the loading conditions of the left ventricle. To avoid such a situation, our study consisted in a direct bilateral intracoronary infusion of enalaprilat in patients with dilated cardiomyopathy and normal coronary arteries (mean ejection fraction = 32 +/- 11%, n = 12). This intracoronary infusion (0.05 mg min-1, 1 ml min-1 in each coronary artery) resulted in no significant change of the systemic resistances (20.6 +/- 5.6 to 22.0 +/- 5.1 mmHg l-1 min), rate-pressure product (10,974 +/- 2630 to 10,214 +/- 2486) or myocardial oxygen consumption (21.08 +/- 6.37 to 22.10 +/- 6.42 ml min-1). Despite these steady haemodynamic conditions, intracoronary enalaprilat provoked a significant elevation of coronary sinus blood flow (181 +/- 73 to 214 +/- 79 ml min-1, P less than 0.001) with a reduction of coronary resistance (0.51 +/- 0.17 to 0.41 +/- 0.15 mmHg ml-1 min, P less than 0.001), and no significant alteration in plasma renin activity or plasma aldosterone. The results of this intracoronary infusion of enalaprilat demonstrate that this angiotensin converting-enzyme inhibitor has significant coronary vasodilator properties, which can be evidenced without stimulating the peripheral renin-angiotensin system.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559851 TI - Co-operation between viral oncogenes in avian erythroid and myeloid leukaemia. PMID- 2559849 TI - Alterations of beta-adrenoceptor mediated relaxations of atherosclerotic human coronary arteries. AB - Preliminary investigations suggest that atherosclerosis attenuates the beta adrenergic relaxations of human coronary arteries. However, this finding could not be related specifically to the beta-agonists and responses to non-adrenergic relaxants could be also decreased. Therefore segments of coronary arteries were isolated from 35 human hearts (aged 18-58 years) with various degrees of atherosclerosis. On rings precontracted with KCl (15 mM) concentration-response curves were constructed for isoproterenol and were compared with the responses to three other vasodilators: forskolin, nitroglycerin and SIN1 (the active metabolite of molsidomine). At the end of each experiment, the degree of atherosclerosis was scored by histological examination. Moderate (class II) and severe (class III) lesions of atherosclerosis significantly altered the maximal response to isoproterenol, which was decreased from 76 +/- 5% (of KCl-induced contraction) to 33 +/- 6% in class II and 22 +/- 5% in class III. Conversely, the maximal relaxations to forskolin, nitroglycerin and SIN1 were not significantly modified. However, in class III, there was a significant shift to the right of the dose-response curves to nitroglycerin and SIN1 (but not to forskolin). Thus our results show that atherosclerosis markedly attenuates the relaxations of human coronary smooth muscles to isoproterenol, and that this abnormality is not related to an alteration in the intrinsic capacity of the atherosclerotic vessel to relax. PMID- 2559848 TI - Quinolone antimicrobial agents: adverse effects and bacterial resistance. AB - Adverse effects, drug-drug interactions and bacterial resistance to the new quinolone antimicrobial agents are reviewed. Clinical adverse effects are reported to occur in 5-10% of patients, and include primarily gastrointestinal disturbances, central nervous system toxicity and rash. Laboratory abnormalities are reported to occur in 5-12% of patients, and include mild reversible elevations of transaminases. Quinolones are not recommended in persons whose bone growth is incomplete or in pregnant or nursing women because cartilage toxicity has been observed in juvenile beagles. Drug-drug interactions may occur between quinolones and theophylline, caffeine, and magnesium- or aluminium-containing compounds such as antacids and sucralfate. Bacterial resistance occurs by chromosomal mutations which alter the target enzyme DNA gyrase or decrease drug accumulation. Emergence of resistance during therapy is uncommon to date but can be problematic in infections with Pseudomonas aeruginosa. Staphylococcus aureus and other bacteria for which the therapeutic index may be low. In summary, quinolones thus far have been well tolerated, but more experience is needed to determine the exact nature and extent of adverse effects and emergence of bacterial resistance. PMID- 2559852 TI - The effect of cytomegalovirus infection on the adherence of polymorphonuclear leucocytes to endothelial cells. AB - Adherence of polymorphonuclear leucocytes (PMN) to the endothelial lining of blood vessels is an essential component of the inflammatory response. In this study the effect of cytomegalovirus (CMV) infection on the adherence of PMNs has been examined using an in vitro model system. Human umbilical venous endothelial cells (HUVEC) were grown on fibronectin-coated plastics. CMV infection of HUVEC resulted in the appearance of viral antigens in a small percentage of the cells. At 24 h post-infection when no virus-induced cytopathic effect could be observed in the cell monolayers, the adherence of PMNs was significantly increased. The virus-induced adherence effect was cell bound and could not be induced by soluble components in the medium of the virus-infected cells. The augmentation of the PMN adherence to CMV-infected endothelium was sensitive to tunicamycin suggesting that the virus infection induces the expression of glycoproteins on the HUVEC membranes which are responsible for the PMN adherence. Thus CMV infection of the endothelium results in an increased adherence of PMNs. In the in vivo situation systemic viral infection can potentially lead to infection of blood vessel endothelium and thus can induce a damage of endothelium. This phenomenon could play a role in the atherogenesis process. PMID- 2559853 TI - Effect of calcium on endothelium-derived relaxing factor formation and cGMP levels in endothelial cells. AB - Various stimulants of the release of EDRF (endothelium-derived relaxing factor) increased intracellular cGMP levels in bovine aortic endothelial cells. ATP was the most effective compound tested, increasing cGMP 7-fold, followed by the calcium ionophore, A23187 (4.8-fold), and bradykinin (4.0-fold). The EC50 values were similar to those obtained when EDRF release was measured with the bioassay technique, which suggests a stimulation of endothelial guanylate cyclase by EDRF. The direct acting stimulants of soluble guanylate cyclase, sodium nitroprusside and SIN-1 (3-morpholino-sydnonimine), also increased the cGMP content of endothelial cells by 9.4 and 7.2 times, respectively. The effects of both groups of stimulants on cGMP levels were antagonized by the lipoxygenase inhibitor, nordihydroguaiaretic acid, and by the radical scavenger, phenylbutylnitrone, whereas gossypol or canavanine only antagonized the EDRF-induced effect on endothelial cGMP levels. Bradykinin, ATP and A23187 also increased the uptake of 45CaCl2 into endothelial cells but since the complete removal of extracellular Ca2+ or blockade of Ca2+ transport by LaCl3 did not affect the ability of these compounds to elevate cGMP levels, the formation of EDRF appears not to be triggered by an influx of extracellular calcium. This study provides evidence that EDRF stimulators enhance cGMP levels in endothelial cells, probably due to a direct activation of guanylate cyclase by EDRF. PMID- 2559854 TI - Chronic opioid antagonist treatment: assessment of receptor upregulation. AB - Changes in specific brain opioid binding and opioid pharmacodynamics were determined in mice treated with the opioid antagonist naltrexone (subcutaneously implanted pellets) for 8 days. Chronic opioid antagonist treatment increased the number of binding sites (upregulation) for [3H]naloxone (+55%) and [3H][D-Ala2, D Leu5]enkephalin (+41%) but did not alter the affinity of the ligands, as determined in saturation studies. Displacement studies of [3H]naloxone by morphine also indicated that there was no change in morphine's affinity. In vivo estimation of naloxone affinity (pA2), agreed with the in vitro results indicating that chronic naltrexone treatment did not alter naloxone affinity. Chronic naltrexone treatment (0.5, 1.0, 15.0 mg pellets) increased the analgesic potency of morphine (supersensitivity) in a dose-dependent manner, up to a maximal increase in relative potency of 1.8. However, in mice tested with the naltrexone pellets still implanted, the 15 mg naltrexone pellet was able to shift the dose-response function for morphine analgesia more than 300-fold. The lowest dose naltrexone pellet (0.5 mg), produced significant antagonism of morphine analgesia, but did not produce significant supersensitivity. Thus, supersensitivity and upregulation are not proportional to the degree of antagonism of opioid effects; and supersensitivity in the mouse is related to increased binding sites and not to changes in receptor affinity as determined by in vivo and in vitro methods. PMID- 2559855 TI - Effects of the novel antidepressant S-adenosyl-methionine on alpha 1- and beta adrenoceptors in rat brain. AB - alpha 1- and beta-adrenoceptors were studied ex vivo in the brains of rats receiving repeated daily treatment with the standard antidepressant imipramine or the atypical antidepressant S-adenosyl-L-methionine (SAM), which has minimal effects on monoamine reuptake or turnover. Consistent with past studies, a decrease in the density of beta receptors at three weeks and an increase in the affinity of alpha 1 receptors for the agonist phenylephrine at one week of treatment was observed with imipramine. By comparison, an increase in the density of beta receptors and a decrease in the affinity of alpha 1 receptors for phenylephrine was observed at one week of treatment with SAM. These changes were no longer apparent at three weeks of treatment. The results suggest that treatment with SAM does lead to changes in adrenergic neurotransmission, but that down regulation of beta receptors or increased agonist affinity of alpha 1 receptors may not be necessary for the production of antidepressant effects. PMID- 2559857 TI - In vivo presynaptic modulation of serotonergic neurotransmission in the rat hippocampus by diazepam. AB - The effect of the benzodiazepine agonist, diazepam, on serotonergic (5-HT) neurotransmission was assessed in in vivo electrophysiological experiments. Diazepam enhanced, in a dose-dependent manner (0.05-2 mg/kg i.v.), the effect of electrical stimulation of the ascending 5-HT pathway on the firing activity of dorsal hippocampus pyramidal neurons. This effect was blocked by the benzodiazepine antagonist, flumazenil. Diazepam did not modify the efficacy of microiontophoretic application of 5-HT and GABA. The results indicate that the activation of benzodiazepine receptors facilitates of 5-HT neurotransmission, presumably through a presynaptic modulatory mechanism. PMID- 2559856 TI - Binding in vivo of selective mu and delta opioid receptor agonists: opioid receptor occupancy by endogenous enkephalins. AB - The in vivo binding properties of cerebral mu and delta opioid receptors were investigated in mice after the intrastriatal injection of [3H][D-Ala2, MePhe4, Gly-ol5]enkephalin (DAGO) or [3H][D-Thr2,Leu5]enkephalyl-Thr (DTLET). Both peptides exhibited similar diffusion kinetics in the brain and 30-40% of [3H]DAGO or [3H]DTLET was shown to be present in the tissue 15 min after injection when maximal binding was observed. The specific binding of both agonists, defined as the fraction of the radioactivity bound to brain membranes which was displaced by 10 nmol of cold ligand, was reversible, saturable and displayed a pharmacological profile similar to that found in in vitro experiments. At doses producing a similar analgesic effect in the hot-plate test in mice, DTLET occupied 64% of delta sites and DAGO 15% of mu sites. However, because of the residual cross reactivity of DTLET for mu sites, it appeared that both ligands occupied a similar number of mu receptors at their ED50 values, thus supporting a preferential involvement of mu opioid binding sites in the supraspinal pain control. [Met5]enkephalin inhibited the in vivo binding of both agonists only when the peptide was protected from degradation by the co-administration of a mixed inhibitor of enkephalin degrading enzymes RB38A (N[3(R)(hydroxyaminocarbonyl)-2-benzyl-1-oxopropyl]- L-phenylalanine). Unlike thiorphan, 5 nmol RB38A alone was able to inhibit [3H]DAGO binding by 60%. This result is the first direct demonstration of the existence of an in vivo tonic control of mu opioid receptor occupation by endogenous opioid peptides. PMID- 2559858 TI - [Work hygiene in greenhouses]. PMID- 2559859 TI - Herpes simplex virus--induced destructive corneal disease. PMID- 2559860 TI - Epithelial electrolyte transport: interrelationships between H+, HCO3- and Cl-. PMID- 2559861 TI - Structural and functional organization of the Na+, K+-pump. PMID- 2559862 TI - The plasma membrane calcium pump: structure, function and regulation. PMID- 2559863 TI - Molecular sieving, receptor processing and peptidolysis as major determinants of peptide pharmacokinetics in vivo. PMID- 2559865 TI - Content of adenine nucleotide translocator mRNA in insulin-producing cells of different functional states. AB - This study was undertaken to characterize the expression of the gene coding for the adenine nucleotide translocator (ANT) in the insulin-producing beta-cell and to study any possible relationship between its expression and the functional state of the beta-cell. Adult and fetal rat pancreatic islets were prepared and cultured under different conditions in vitro. The total RNA from these islets and from the insulin-producing RINm5F cells was isolated and analyzed by the Northern blot technique via a cDNA clone (pAAC-9) coding for the bovine ANT. We found that a 1600-base pair (bp) mRNA hybridizing to the pAAC-9 clone could be detected in RINm5F cells, and a 1450-bp mRNA was similarly observed in the islets. These sizes correspond well to previously reported forms of mRNA for the ANT observed in other tissues. When comparing the intensities of the pAAC-9 hybridizing bands of the different islet groups, it was observed that fetal islets contained less of this mRNA than adult islets. Furthermore, the content of the ANT mRNA in adult islets cultured at a high glucose concentration was increased compared with islets cultured at a low glucose concentration. Finally, streptozocin-treated islets, which display an impaired glucose-sensitive insulin release after 6 days in culture, also contained less of this mRNA than the control islets. We conclude that pancreatic islet cells express an mRNA that appears to be highly homologous to the bovine ANT and that the contents of this mRNA increases with the functional status of the beta-cell. It is furthermore suggested that defects in the expression of this gene may be associated with impaired glucose sensitivity. PMID- 2559864 TI - Postreceptor effects of sulfonylurea on skeletal muscle glycogen synthase activity in type II diabetic patients. AB - To elucidate the subcellular mechanism of action of sulfonylurea on glucose utilization of skeletal muscle, we studied nine newly diagnosed patients with type II (non-insulin-dependent) diabetes. Examinations were performed before and after 8 wk of gliclazide therapy. Gliclazide treatment was associated with improved glycemic control and enhanced pancreatic beta-cell responses to meal stimulation. During euglycemic insulin clamps, insulin-inhibited endogenous glucose production was improved after gliclazide therapy. Moreover, mean (+/- SE) glucose disposal rate increased from 3.2 +/- 0.7 to 4.8 +/- 0.8 and from 7.9 +/- 0.9 to 10.4 +/- 0.9 mg.kg-1.min-1 at in vivo plasma insulin levels of approximately 75 and approximately 320 mU/L, respectively. In addition, insulin receptor function and glycogen synthase activity were analyzed in skeletal muscle biopsies obtained in seven patients. The biopsies were obtained during basal insulinemia and hyperinsulinemia (approximately 320 mU/L) before and after treatment. Insulin receptors purified with wheatgerm agglutinin showed unchanged insulin-binding properties and unchanged receptor kinase function with respect to basal and insulin-stimulated phosphorylation of exogenous peptide poly(Glu80Tyr20). Gliclazide treatment had no effect on the maximal activities of glycogen synthase. Moreover, in biopsies obtained at basal insulinemia, the half maximal activation constant for glucose 6-phosphate (A0.5) was identical before and after therapy (0.54 +/- 0.05 vs. 0.54 +/- 0.05 mM, respectively, NS). However, in biopsies obtained at hyperinsulinemia, A0.5 was 0.30 +/- 0.05 vs. 0.20 +/- 0.02 mM before and after gliclazide therapy, P less than .04.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559866 TI - Effects of high glucose on insulin secretion by isolated rat islets and purified beta-cells and possible role of glycosylation. AB - We investigated the effect of 24 h of exposure to various glucose concentrations on insulin secretion by isolated rat pancreatic islets and purified rat beta cells. Compared with islets cultured with standard medium (5.5 mM glucose), islets cultured with 16.7 mM glucose showed a higher basal insulin release (means +/- SE, 3.0 +/- 0.5 vs. 0.7 +/- 0.2%, n = 8, P less than .005) and reduced glucose-stimulated insulin secretion (2.4 +/- 0.3 vs. 5.8 +/- 0.4%, n = 8, P less than .005). Similar results were also obtained with purified beta-cells. The effect of high glucose was time dependent (present after 12 h, maximal after 24 h) and reversible: when islets cultured with high glucose were transferred to standard medium, normal responsiveness to glucose was restored within 8 h and normal basal release within 24 h. Mannitol, 3-O-methylglucose, and 2-deoxyglucose were not able to mimic the effects of glucose. Islets or purified beta-cells cultured in the presence of high glucose had a normal response when stimulated with glyburide, dibutyryl cyclic AMP, and isobutylmethylxanthine. Tunicamycin, an inhibitor of N-terminal glycosylation, prevented glucose-induced desensitization when added during 24 h of islet culture with 16.7 mM glucose. Swainsonine, another agent that influences glycosylation, had a similar effect. Our study indicates 1) that 24 h of exposure to high glucose induces a specific and reversible impairment of insulin secretion in response to glucose, 2) that this is a direct effect of glucose on beta-cells, and 3) that islet glucose metabolism and glycosylation processes may play a critical role in determining glucose desensitization. PMID- 2559867 TI - Prevention of nerve edema and increased blood-nerve barrier permeability-surface area product in galactosemic rats by aldose reductase or thromboxane synthetase inhibitors. AB - Nerve water content and the permeability-surface area product (PA) to [3H]-or [14C]sucrose at the blood-nerve barrier were determined in unanesthetized control rats fed a normal diet and in rats fed galactose with or without an aldose reductase inhibitor (Statil or AL 1576) or a thromboxane synthetase inhibitor (CGS 12970). Nerve water content was determined by taking the difference between dry and wet weights of whole tibial nerves. PA was determined by an intravenous bolus injection of radiotracer with multiple-time-point graphic and quantitative autoradiographic methods. The mean nerve water content in galactosemic rats was 15% higher than in control rats after 7-11 mo on the diet. Statil and AL 1576 prevented nerve edema, but CGS 12970 was only partially effective in preventing an increase in nerve water content in galactose-fed rats. In galactosemic rats, the mean PA to sucrose at the blood-nerve barrier, calculated from nerve dry weight, was twofold higher than in control rats. Treatment with Statil, AL 1576, or CGS 12970 prevented increased PA. Our results suggest that nerve edema and increased blood-nerve barrier PA are secondary to polyol production and can be prevented by inhibiting aldose reductase. PMID- 2559868 TI - Isolation and characterization of a multipotent clone of human embryonal carcinoma cells. AB - Histopathological studies suggest that the stem cells of human teratomas may be classified into two major categories: nullipotent stem cells, and multipotent stem cells, capable both of self-renewal and differentiation into a wide range of somatic and extraembryonic cell types. We have isolated a multipotent stem cell clone from the human teratoma cell line GCT 27, and compared its properties to a nullipotent clone derived from the same strain. The multipotent clone GCT 27 X-1 gave rise to colonies of mixed cell morphology in vitro. Analysis of cell surface, cytostructural and extracellular matrix markers in GCT 27 X-1 cells showed that the stem cells of this line were very similar in phenotype to nullipotent cells. The two cell clones were predominantly hypotriploid, and contained several marker chromosomes in common. GCT 27 X-1 was feeder-cell dependent for continuous growth in vitro; removal of the feeder layer resulted in differentiation of the stem cells into a variety of cell types, some with characteristics of extraembryonic endoderm, others showing neuronal properties. When transplanted into nude mice, GCT 27 X-1 cells gave rise to teratocarcinomas containing embryonal carcinoma stem cells, and many other cell types: yolk sac carcinoma cells; cells producing alphafetoprotein or human chorionic gonadotrophin; glandular, columnar, cuboidal, and squamous epithelium; primitive mesenchyme and cartilage; neuroectodermal cells. Nullipotent GCT 27 C-1 cells could form colonies in the absence of feeder layers, but multipotent GCT 27 X-1 cells could not. While a range of known growth factors and related substances failed to substitute for feeder layers in supporting the growth of GCT 27 X-1 stem cells, supernatants from yolk sac carcinoma cell line GCT 44 could partially replace the feeder cell requirement. Thus, the results revealed a basic difference in growth control between these multipotent and nullipotent human embryonal carcinoma cells, and suggested a possible paracrine regulatory pathway between multipotent stem cells and yolk sac carcinoma cells. PMID- 2559869 TI - [Genetic instability and transposition of mobile element mdg4 in a mutator strain of Drosophila melanogaster]. AB - Laboratory mutator strain of Drosophila melanogaster is characterized by increased (up to 10(-3)-10(-4) frequency of spontaneous mutability. Mutations appear in premeiotic stages of gametes development. The majority of mutations were unstable (high frequencies of reversions, appearance of new mutations at the same and other loci, replicating instability). Localization of mobile elements mdg1, mdg2, mdg3, mdg4, copia and P element in X chromosomes of mutator individuals and its mutations y, ct, sbt was studied by hybridization in situ. In all strains P element was absent. The distribution of mdg1, mdg2, mdg3 and copia was identical in mutator strains and its derivatives, but distribution of mdg4 was different. The essential heterogeneity in localization of mdg4 and increased (up to 30-40) copy number in the mutator strain individuals was observed. The ability of single element mdg4 to autonomous transpositions was thus shown. PMID- 2559870 TI - [Characteristics of inserted mutations obtained in the P-M hybrid dysgenesis system in the 9F12-10A7 region of the Drosophila melanogaster X-chromosome]. AB - 19 new mutations in the 9F12-10A7 region of Drosophila melanogaster X chromosome was obtained in the system of P-M hybrid dysgenesis. They appeared to be lethals, as judged from viability of homo- or hemizygous females. In situ hybridization of P DNA with polytene chromosomes revealed P-element insertion in the 10A1-2 band in the majority of the mutants. As a result of complementation analysis, all these mutations were localized at previously known loci: l(1)BP1, l(1)BP5, l(1)BP8, l(1)BP7. No insertion mutations were found at the vermilion locus. This can imply for non-random distribution of insertion mutations in the region studied. Further comparison of these mutations with previously EMS-induced ones revealed that insertion mutations are predominantly hypomorph lethals which do not influence the viability, morphology and fertility of homozygous males and females, but drastically reduce viability of hemizygous females. PMID- 2559871 TI - Hybrid dysgenesis-induced response to selection in Drosophila melanogaster. AB - In Drosophila melanogaster, the P-M and I-R systems of hybrid dysgenesis are associated with high rates of transposition of P and I elements, respectively, in the germlines of dysgenic hybrids formed by crossing females of strains without active elements to males of strains containing them. Transposition rates are not markedly accelerated in the reciprocal, nondysgenic hybrids. Previous attempts to evaluate the extent to which hybrid dysgenesis-mediated P transposition contributes to mutational variance for quantitative characters by comparing the responses to selection of P-M dysgenic and nondysgenic hybrids have given variable results. This experimental design has been extended to include an additional quantitative trait and the I-R hybrid dysgenesis system. The selection responses of lines founded from both dysgenic and nondysgenic crosses showed features that would be expected from the increase in frequency of initially rare genes with major effects on the selected traits. These results differ from those of previous experiments which showed additional selection response only in lines started from dysgenic crosses, and can be explained by the occasional occurrence of large effect transposable element-induced polygenic mutations in both dysgenic and nondysgenic selection lines. High rates of transposition in populations founded from nondysgenic crosses may account for the apparently contradictory results of the earlier selection experiments, and an explanation is proposed for its occurrence. PMID- 2559874 TI - Construction and characterization of versatile kanamycin-resistance cassettes derived from the Tn5 transposon. AB - We have developed plasmids with the Tn5 kanamycin-resistance gene (kan) flanked either symmetrically or asymmetrically by several restriction sites. These can be used to provide a selectable genetic marker or to mobilize restriction sites and sense or nonsense codons into genes. The 1.3-kb kan cassette exhibits polarity effects in both directions. PMID- 2559872 TI - U2 snRNA sequences that bind U2-specific proteins are dispensable for the function of U2 snRNP in splicing. AB - Previously we showed that microinjection of purified U2 snRNA from HeLa cells into Xenopus laevis oocytes, depleted of their endogenous U2 snRNPs by oligonucleotide-targeted degradation, led to assembly of hybrid snRNPs that were fully functional for splicing of SV40 late pre-mRNA. We have extended these results by examining features of U2 RNA that are required for its role in splicing. Injection of Xenopus U2 snRNA transcribed in vitro by T7 RNA polymerase, differing in sequence from authentic U2 by only one nucleotide, although capable of efficient assembly into snRNP-like particles, did not complement U2-predepleted oocytes for splicing. However, when injected into pretargeted oocytes, a plasmid containing Xenopus U2 snRNA sequences resulted in synthesis of U2 snRNA that was assembled into snRNPs capable of mediating splicing of SV40 late pre-mRNA. This allowed us to test several U2 RNA mutants for their function in splicing. Mutants with sequences deleted within U2 stem loops I and II, although efficiently assembled into snRNP-like particles upon injection, failed to restore splicing. Interestingly, however, injection of a mutant that lacks the binding site for the U2-specific proteins A' and B", restored pre-mRNA splicing. These data suggest that the direct binding of U2 specific proteins with snRNA is not essential for the function of U2 snRNPs in splicing of pre-mRNA. PMID- 2559873 TI - Induction of transcription factor AP-1 by adenovirus E1A protein and cAMP. AB - Treatment of adenovirus-infected mouse S49 cells with cAMP analogs leads to the transcriptional induction of early viral genes. E1A proteins and cAMP work in synergy to activate several of these genes. We now demonstrate that the transcription factor AP-1 is modestly induced by cAMP in S49 cells and induced to significantly higher levels by cAMP in the presence of E1A proteins. Cytoplasmic levels of c-fos and junB mRNAs are rapidly increased by cAMP, and the induction is substantially stronger in the presence of E1A protein. The AP-1 activity binds efficiently to both AP-1 and activating transcription factor (ATF)/cAMP response element binding protein (CREB)-binding sites present in E1A-inducible promoters and presumably plays a role in the transcriptional activation of adenovirus genes by E1A proteins and cAMP. PMID- 2559875 TI - Development of a shuttle vector and a conjugative transfer system for Actinobacillus pleuropneumoniae. AB - An original genetic system for Actinobacillus pleuropneumoniae has been developed. A shuttle cloning vector, pYG53, was constructed from the wild-type plasmid pYG10. It permits, in conjunction with electroporation, the introduction of cloned genes into this species. A conjugal transfer system between Escherichia coli and A. pleuropneumoniae involving pYG54, a mobilizable derivative of pYG53, is also described. Conjugation efficiencies of 8.3 x 10(-3) exconjugants per donor can be obtained. PMID- 2559876 TI - Sequence analysis of the chromosomal and plasmid genes encoding phosphoribulokinase from Alcaligenes eutrophus. AB - Two DNA fragments encoding the chromosomal and plasmid copies of the gene (cfxP) encoding phosphoribulokinase (PRK) from the chemoautotrophic bacterium Alcaligenes eutrophus, were sequenced and found to be highly homologous. The gene (cfxF) of another Calvin cycle enzyme, fructose-1,6-bisphosphatase (FBPase), was identified as terminating immediately upstream of cfxP, but was not completely contained on both fragments. A hypothetical, also incompletely contained, open reading frame starts closely downstream from cfxP. Genes cfxF, cfxP, and the third hypothetical gene seem to belong to the same operon. The cfxP genes encode highly homologous PRK isoenzyme subunits consisting of 292 aa residues with calculated Mrs of 33 319 (chromosomal PRKc) and 33 164 (plasmid-encoded PRKp). There is little overall sequence similarity between the bacterial and plant (spinach) PRK, apart from some structural motifs. PMID- 2559877 TI - Sumo15A: a lambda phasmid that permits easy selection for and against cloned inserts. AB - We report the construction of a phasmid vector, Sumo15A, designed for recombination-based screening of recombinant DNA libraries [Seed, Nucleic Acids Res. 11 (1983) 2427-2445]. This vector permits rapid selection in Escherichia coli for homology-mediated integration and excision between homologous DNA inserts cloned in a supF-carrying plasmid and in Sumo15A. The region available for recombination spans the homologous sequence shared by the plasmid and the phasmid. SupF is the selection tool that we used. Efficient selection for supF expression by Sumo15A requires recombination mediated by the lambda phage red gene, which promotes homologous recombination between phage and plasmid DNAs. Counterselection against supF expression by Sumo15A occurs because the presence of a pSC101-derived plasmid replicon in this phasmid permits the growth of Sumo15A as a plasmid in a specialized host, E. coli strain DK37. In strain DK37, Sumo15A cannot replicate as a phage, and the presence of a plasmid-carrying supF is lethal to cells plated on galactose plates. This scheme was developed to select for sequences that are transcribed from chromosomes of interest. PMID- 2559878 TI - Saturation mutagenesis of the inside end of insertion sequence IS50. AB - A 19-bp segment at the inside (I) end of IS50 (Tn5) is needed for efficient transposition. The importance of each position was assayed by making at least one base substitution at each position by either chemical-or oligodeoxyribonucleotide directed mutagenesis. Mutant I ends were paired with a wild-type (wt) segment from the outside (O) end of IS50 and the transposase (tnp) gene was placed either between the ends or 1200 bp from the O end. The frequency of transposition of the resultant elements to bacteriophage lambda was measured. At least one substitution at each of the 19 I-end positions decreased transposition activity to less than 25% of wt, and most substitutions (25 of 28) decreased it to less than 5% of wt from one or both donor plasmids. These results show that each position in the I end is important during transposition. PMID- 2559879 TI - Transposable elements for efficient manipulation of a wide range of gram-negative bacteria: promoter probes and vectors for foreign genes. AB - We describe here the construction and use of a series of modified transposons based on the insertion sequence IS1. Like their parent, omegon-Km [Fellay et al., Gene 76 (1989) 215-226], these elements permit efficient insertional mutagenesis of a variety of Gram-negative bacteria. The presence of a functional pBR322 origin of replication within the transposable element facilitates subsequent cloning of the mutated gene. The omegon-Km system was previously shown to function in Pseudomonas putida, Rhizobium leguminosarum and Paracoccus denitrificans. The results we present here demonstrate that its use can be extended to Xanthomonas campestris, a plant pathogen, and to the microaeroduric Zymomonas mobilis. Derivative transposons carrying unique restriction sites for ScaI, NdeI, XbaI and XhoI have been constructed, allowing the cloning and introduction of foreign genes. We have also constructed two derivatives which can be used to generate operon fusions upon insertion and are thus useful for isolating and characterising indigenous promoters. One carries a promoterless chloramphenicol acetyl-transferase (CAT)-encoding gene (cat) and the second, the entire promoterless Escherichia coli lac operon. We demonstrate the utility of the cat promoter probe in X. campestris to target conditional promoters inducible by high salt or subject to repression by glucose. PMID- 2559880 TI - Semisynthetic promoters activated by cyclic AMP receptor protein of Escherichia coli. AB - Semisynthetic promoters activated by Escherichia coli cyclic AMP receptor protein (CRP) were created by combining a synthetic CRP-binding site (crb) and nucleotide sequences derived from cryptic promoter regions. A 22-bp oligodeoxyribonucleotide corresponding to an idealized crb was randomly placed into DNA regions that precede a promoterless lacZ gene on a plasmid. Several plasmid clones were obtained which allowed the expression of lacZ in crp+ cya+ cells carrying a chromosomal deletion of lac genes. The beta-galactosidase and the quantitative S1 nuclease assays of crp+ and delta crp cells harboring these plasmids indicated that the transcription from newly created promoters is dependent on CRP. Sequence analysis revealed that these promoters are divided into two types based on the location of the crb relative to the transcription start point (tsp). The distance from the center of the crb to the tsp is 70 bp in the first type and 38 bp in the second type. The sequences of all these promoters exhibit poor homology with the consensus promoter sequence. PMID- 2559881 TI - Self-reduction of the iron(III)-doxorubicin complex. AB - The Fe3(+)-doxorubicin complex undergoes reactions that suggest that the complex self-reduces to a ferrous oxidized-doxorubicin free radical species. The Fe3(+) doxorubicin system is observed to reduce ferricytochrome c, consume O2 and react with 2,2'-bipyridine. Bipyridine acts as a "ferrous ion scavenger" as it reacts with the ferrous ion produced by Fe3(+)-doxorubicin self-reduction. In the absence of O2, a ferrous doxorubicin complex accumulates. In the presence of oxygen, Fe2+ recycles back to Fe3+. The rates of these reactions were measured and the Fe3(+)-doxorubicin self-reduction was determined to be the rate determining step. The Fe3(+)-doxorubicin induced inactivation of cytochrome c oxidase and NADH cytochrome c reductase on beef heart submitochondrial particles occurs at a rate similar to Fe3(+)-doxorubicin self-reduction. Thus the rate at which damage to these mitochondrial enzymes occurs may be controlled by a nonenzymatic Fe3(+)-doxorubicin self-reduction. PMID- 2559882 TI - The inhibition of catalase by glutathione. AB - Reduced glutathione (GSH) inhibited catalase activity in a dose-dependent manner. DL-dithiothreitol (DL-DTT) and dithioerythritol (DTE) also inhibited catalase activity. The inhibition of catalase by GSH and DL-DTT could be reduced by NADPH. Polyacrylamide gel electrophoresis demonstrated the inhibition was partially reversible. The inhibition of catalase by GSH appeared to be partly due to superoxide radicals, since it was inhibited by active manganese superoxide dismutase, but not by heat-inactivated enzyme. Other chemical species also appear to take part in the inhibition, but they could not be identified. PMID- 2559883 TI - The interaction between nitrogen oxides and hemoglobin and endothelium-derived relaxing factor. AB - Among nitrogen oxides, NO and NO2 are free radicals and show a variety of biological effects. NO2 is a strongly oxidizing toxicant, although NO, not oxidizing as NO2, is toxic in that it interacts with hemoglobin to form nitrosyl- and methemoglobin. Nitrosylhemoglobin shows a characteristic electron spin resonance (ESR) signal due to an odd electron localized on the nitrogen atom of NO and reacts with oxygen to yield nitrate and methemoglobin, which is rapidly reduced by methemoglobin reductase in red cells. NO was found to inhibit the reductase activity. Part of NO inhaled in the body is oxidized by oxygen to NO2, which easily dissolves in water and converts to nitrite and nitrate. The nitrite oxidizes oxyhemoglobin autocatalytically after a lag. The mechanism of the oxidation, particularly the involvement of superoxide, was controversial. The stoichiometry of the reaction has now been established using nitrate ion electrode and a methemoglobin free radical was detected by ESR during the oxidation. Complete inhibition of the autocatalysis by aniline or aminopyrine suggests that the radical catalyzes conversion of nitrite to NO2, which oxidizes oxyhemoglobin. Recently NO was shown to be one of endothelium-derived relaxing factors and the relaxation induced by the factor was inhibited by hemoglobin and potentiated by superoxide dismutase. PMID- 2559884 TI - Mode of delivery and lymphocyte beta 2-adrenoceptor density in parturients and newborns. AB - Twenty-six women with uncomplicated pregnancy at term were selected for the study. Nine had spontaneous vaginal delivery without medication, 8 elective caesarean section under epidural and 9 under general anaesthesia. The results clearly demonstrate that newborns have lower lymphocyte beta 2-adrenoceptor density than their mothers. In vaginally delivered newborns the lymphocyte beta 2 adrenoceptor density was 38%, in the caesarean section group with general anaesthesia 27%, and with epidural anaesthesia 22% lower than in the corresponding mother group. Vaginally delivered newborns have lower lymphocyte beta 2-adrenoceptor density than those delivered by caesarean section. A plausible explanation is the down-regulation of the beta 2-adrenoceptors during labour and delivery. PMID- 2559885 TI - Intratumoral echo-guided injection of interleukin-2 and lymphokine-activated killer cells in hepatocellular carcinoma. AB - Interleukin-2 has proved to be effective for the intralesional treatment of tumors of the bladder. There are examples in literature of hepatocellular carcinoma (HCC) treatment with lymphokine-activated killer (LAK) cells infused in the hepatic artery. We decided to check the effects of echo-guided intralesional injection of these cells in this disease. We treated 5 patients with inoperable hepatocellular carcinoma, following cirrhosis; in 4 cases the mass had a diameter less than 3 cm (small HCC) while in the remaining case it measured 7 cm. Tumor size remained unchanged in 3 of the 4 small HCC, and increased only slightly in the other (over a period of 10 months). This would appear to indicate that treatment halted neoplasm growth or at least slowed it down. The echo pattern of the lesions changed, with a constant reduction in echogenicity. Finally, in multiple control biopsies, fibrosis, present in only one case before treatment, was found fairly constantly after treatment. There were no significant side effects, apart from slight water retention in one patient. On the basis of our preliminary results, we consider it worthwhile continuing this study to establish the most suitable IL-2 doses and analyze in more detail the modifications induced in the neoplasm. PMID- 2559886 TI - Better survival in women than in men after radical resection of hepatocellular carcinoma. AB - During the period between January 1980 and December 1987, 229 male and 39 female adult patients with primary hepatocellular carcinoma (HCC) were hospitalized in our unit. Radical hepatic resection was carried out in 90 (39.3%) males and 17 (43.6%) females in whom no specific cancer treatment had been attempted preoperatively. The 1-, 3-, and 5-year survival rates in the male and female patients were 78% and 70%, 45% and 52%, and 19% and 52%, respectively. The difference was significant after 47 months. No substantial differences were found between the two groups with respect to age, preoperative clinical condition and laboratory data, method of liver resection, postoperative morbidity and mortality, postoperative adjuvant chemotherapy, and histopathology of HCC and the liver. Only the incidence of alcohol abuse was significantly different, being higher in male than in female patients. However, the survival analysis demonstrated that alcohol abuse had had no influence on recurrence rate and long term survival in either male or female patients. As a control, survival was analyzed for the patients with similar clinicopathological background but without HCC who had undergone distal splenorenal shunt for esophageal varices. There was no significant difference between the male and female patients. The current clinical results seems to support our hypothesis based upon sex hormone receptor studies that HCC may be androgen-dependent. PMID- 2559887 TI - Coupling of the enzymic activities of myosin ATPase and creatine kinase and its role in muscular contraction. AB - During muscular contraction the regeneration of ATP, catalysed by creatine kinase (CK), keeps pace with the hydrolysis of ATP by myosin ATPase posing the question of its regulatory mechanism. In the background of F-actin activation of heavy meromyosin (HMM) ATPase activity we have investigated in vitro the role of F actin in regulating CK's activity in the absence and presence of HMM. For the coupled enzyme system we have also looked into the roles played by the individual reactants. F-actin has been found to appreciably increase CK's activity in the absence of HMM. While HMM alone inhibited CK's activity, there was a several fold increase when F-actin was also present. By a process of elimination we conclude that none of the reactants apart from H+ could be involved in regulating CK's activity in the coupled enzyme system. As no change in the pH of reaction mixture was observed during the reaction, we further conclude that the two enzymic reactions are coupled by proton transfer along F-actin. Implications of the findings for PCr-Cr shuttle and movements of ATP and ADP in sarcomere are discussed. PMID- 2559889 TI - Determination of distances of sugar protons from Mn2+ in concanavalin A. AB - 1H NMR spin-lattice relaxation time (T1) measurements have been carried out with various sugars, viz. methyl alpha-D-glucopyranoside (alpha-MeGluP), methyl beta-D glucopyranoside (beta-MeGluP), methyl alpha-D-mannopyranoside (alpha-MeManP), maltose (4-O-alpha-D-glucopyranosyl-D-glucose), nigerose (3-O-alpha-D glucopyranosyl-D-glucose), p-nitrophenyl alpha-maltoside (PNP-alpha-maltoside) and p-nitrophenyl beta-maltoside (PNP-beta-maltoside) to determine the distances of sugar protons from Mn2+ in concanavalin A (Con A). With a rotational correlation time of 1.58 x 10(-10) s determined, distances were calculated using Solomon-Bloembergen equation. The data obtained indicated differences in disposition of different groups in the binding site of Con A. An average value of about 10 A was obtained for the distances of sugar protons from Mn2+ in Con A. In the case of mono and disaccharides, the non-reducing end sugar unit was found to be closer to Mn2+ than the reducing end one. PMID- 2559888 TI - Changes in lipid composition and some biologically important enzyme activities in the microsomal membranes of developing toad ovary in different seasons. AB - The microsomal membranes isolated by sucrose density gradient centrifugation from developing toad ovary have been found to differ significantly in lipid composition and various enzyme activities in different seasons. All the enzymes studied, viz. Na+, K(+)-ATPase, delta 5-3 beta-hydroxysteroid dehydrogenase (delta 5-3 beta HSD) and prostaglandin synthetase, exhibited maximum activity during the breeding season (July-September) at all stages of development (a,b,c & d). The activities of Na+, K(+)-ATPase and delta 5-3 beta HSD increased with development while that of prostaglandin synthetase followed the reverse order. The total phospholipid, cholesterol and fatty acid contents also varied with season and development. The increase in Na+, K(+)-ATPase and delta 5-3 beta HSD activities in the microsomal membranes of toad ovary at breeding season is accompanied with concomitant increase in phospholipid and unsaturated fatty acid contents at different stages in this season, thereby suggesting some correlation between them. PMID- 2559890 TI - In vitro capacitation of goat spermatozoa. AB - Goat epididymal and ejaculated spermatozoa were incubated in Krebs-Ringer bicarbonate buffer containing pyruvate and lactate as energy source. A 3 hr incubation for epididymal and 4 hr for ejaculated spermatozoa was required for the capacitation and acrosome reaction to take place. Calcium is an essential requirement which was needed for motility maintenance/activation and for the initiation of acrosome reaction. A 2-fold increase in cAMP content was measured over 3 hr period of incubation of goat epididymal spermatozoa which was not seen when calcium ions were either omitted or chelated with EGTA. There is thus a definite involvement of Ca2+ ions and cAMP in capacitation and acrosome reaction of goat spermatozoa. PMID- 2559891 TI - In vivo distribution of lithium in plasma and brain. AB - A single administration of LiCl (0.5, 2 and 4 mmol/kg) to adult male albino rats produced a dose dependent increase of Li level in plasma, whole brain and brain regions. The concentration of Li in whole brain and brain regions was much less than that in plasma. Further, it is also found that concentration of Li in plasma reached a peak at 8 hr while that of Li in whole brain and brain regions reached a peak at 12 hr after the administration. The distribution and retention of Li was found to be highest in hypothalamus followed by striatum, pons-medulla, cerebellum and cerebral cortex. Daily administration of LiCl at a dose of 0.5 and 2 mmol/kg/day showed a time and dose dependent increase in plasma Li level up to a period of 21 consecutive days. But at higher dose (4 mmol/kg/day), on the other hand, under similar condition showed a time dependent increase in plasma Li level up to a period of 14 consecutive days and then gradually decreased with prolongation of treatment to 21 consecutive days. In brain there was no such decrease, rather increase in Li level was observed with the prolongation of duration of treatment, highest concentration of Li was found in hypothalamus and striatum than the rest of the brain regions. These results suggest that under short term treatment with LiCl, the clearance rate of Li in brain cell is much slower than that in plasma. Both single and long-term exposure of LiCl produces a dose dependent increase of Li in plasma, whole brain and brain regions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559892 TI - Immune status of volunteers one year after administration of Japanese encephalitis vaccine produced in India. AB - The immune status of 40 volunteers who received the full course of Japanese encephalitis (JE) vaccine a year earlier and 15 individuals who had received only a booster dose at the same time, was studied by estimating the level of persistence of protective antibody in the sera. All the sera showed persistence of 100 per cent seroconversion rate. Individuals who had the full course of vaccination still had high levels of antibody (mean 2.8 Iog10); however there was a fall of 0.8 Iog10 from the post-booster level. Volunteers who had received only a booster dose, also showed persistence of high level of protective antibody (mean 2.4 Iog10), a drop of 0.9 Iog10 from the post-booster level. Neutralizing (N) antibody estimated using Dibrugarh (7812474) strain of JE virus also demonstrated persistence of high level of protective antibody against this virus (mean 2.4 Iog10). Persistence of high level of protective antibody against homologus and heterologus (Dibrugarh) virus strains and absence of vaccine related side-effects even one year after administration of JE vaccine produced in India, demonstrates the immunizing potency and safety of this new vaccine. PMID- 2559893 TI - Effect of silica treatment on histopathology of intestinal & hepatic amoebiasis in murine model. AB - Histopathological changes in intestinal and hepatic amoebiasis were studied in Swiss albine mice with and without silica pretreatment. The intestinal infection rate was higher in silica treated mice (83%) as compared to non-silica treated mice (33%). Histologically all the silica treated mice had caecal ulceration which tended to spread across the muscularis propria into the serosal layer. The hepatic lesions were observed grossly in 100 and 60 per cent mice with and without silica treatment, respectively. However, the liver lesions in the silica treated mice were more severe, leading to satellite abscesses and in some cases the capsule ruptured leading to adhesions to the diaphragm and peritoneum. Histologically, the blockade of histiocytes with silica resulted in mainly a neutrophilic damage which was extensive and widespread. It also destroyed the amoeba and led to abundant fibrosis. In the non-silica group the damage was slow and devoid of neutrophils with quicker healing and lack of fibrosis resembling human hepatic lesions. PMID- 2559894 TI - Chediak-Higashi syndrome. PMID- 2559895 TI - Calcium and sodium transport in gestational hypertension. AB - The enzymatic activities of Na+/K+ ATPase and Ca2+ ATPase were determined on erythrocyte membranes from 9 normotensive and 9 gestational hypertensive pregnant women near term. A reduction in the activity of the Na+/K+ ATPase and a relative increase in the activity of the Ca2+ ATPase were found in the hypertensive patients, possibly due to a conformational alteration of erythrocyte membranes. This observation supports the possible role of the transmembrane cation transport in the pathogenesis of gestational hypertension. PMID- 2559896 TI - Diet and survival of patients with colon cancer in Utah: is there an association? AB - The extent to which diet influenced colon cancer survival was examined in 411 colon cancer cases identified in Utah between 1976 and 1981 using data from two population based case-control studies. After adjustment by proportional hazards regression models for the effects of tumour stage, age, sex, and religion, the hazard rate ratios (HRR) comparing highest to lowest quartile of intake for total calories, fat and protein were 0.60, 0.81 and 0.66 respectively, with the effect of calories being greatest for short-term survival (less than or equal to 24 months), HRR = 0.49. By contrast, the highest quartile of dietary fibre intake was associated with decreased survival (HRR = 1.53) when compared with the lowest quartile. More extensive studies are needed to verify these findings and to identify mechanisms underlying these associations. PMID- 2559897 TI - Iodine supplementation with oral or intramuscular iodized oil. A two-year follow up of a comparative trial. AB - A controlled trial of iodine supplementation comparing oral with intramuscular iodized oil has been carried out in an iodine deficient area of Zaire. Two years after the administration of 2 ml of oral iodized oil to the population of four villages the overall goitre prevalence had fallen from 64 to 54%. In a further two villages given 2 ml of intramuscular oil the prevalence fell from 65 to 50%. The effectiveness of supplementation was also assessed by measuring changes in thyroid function in women of reproductive age. Among women in the villages given oral iodized oil, the geometric mean thyroxine concentration, measured in dried bloodspots, rose from 27.2 to 52.6 nmol/L at the two-year follow-up. This was similar to the response of the intramuscularly treated villages in which thyroxine levels rose from 32.1 to 65.4 nmol/L. There was no change in goitre prevalence or thyroid function in two control villages. Oral iodized oil is a cheaper and simpler alternative to the injected form providing effective iodine prophylaxis for up to two years after a single dose. PMID- 2559898 TI - [What is assured in drug prevention of thrombosis with special reference to low molecular weight heparin?]. PMID- 2559899 TI - [Aortocaval fistula in ulcerative colitis and primary sclerosing cholangitis]. PMID- 2559900 TI - Use of simian immunodeficiency viruses for AIDS research. AB - Despite frequent statements to the contrary, there are good animal models for AIDS. In this review, we summarize the properties of one of the most useful animal models: infection of rhesus monkeys with simian immunodeficiency virus (SIV). The SIVs are an extensive group of HIV-related lentiviruses of nonhuman primates. They closely resemble the human AIDS viruses, HIV-1 and HIV-2, in both genetic sequence and biological properties. Some SIV isolates, most notably those derived from macaques and mangabeys, induce AIDS in rhesus monkeys in a time frame suitable for laboratory investigation. Rhesus monkeys are not endangered in the wild, they breed well in captivity, and they are available in reasonably large numbers. Study of SIV has already resulted in seminal contributions regarding the origins of the HIVs, AIDS pathogenesis, and vaccine and therapy research. Continued use of SIV systems will be an important weapon in our arsenal against AIDS. PMID- 2559901 TI - Preliminary characterization of a group of actinophages of the thermophilic actinomycete genus Saccharomonospora. AB - The comparison of the Saccharomonospora phages phi SaC1, phi SaG1, phi SaV1, phi SaV2, phi SaV3, and Tm1 demonstrated that they all belong to a single group of closely related actinophages: they were similar with regard to host range, virion mophology (B1 type), genome length (43-45 kb), and GC content of their genomes (63, 3% G + C). Furthermore, DNA-DNA hybridization showed that the phage genomes had a high degree of homology to each other. The phages are, therefore, proposed as representatives of a new bacteriophage species in the sense of the ICTV. PMID- 2559902 TI - Sites of Epstein-Barr virus replication in acute and chronic active Epstein-Barr virus infections. AB - We examined Epstein-Barr virus (EBV) in saliva samples and peripheral blood mononuclear cells (PBMC) from 9 patients with acute infectious mononucleosis (IM) and 4 patients with chronic active EBV infection (CEBV). All saliva samples from patients with acute IM were positive for EBV by both Southern blot hybridization assay and the cord transformation assay. In contrast, EBV DNA could not be detected in saliva samples from patients with CEBV. Only one of the saliva samples from 4 patients with CEBV could transform cord blood lymphocytes with extreme difficulty. No saliva samples from patients with CEBV were capable of inducing early antigens in Raji cells. On the other hand, EBV DNA was detected in PBMC from 3 of the 4 patients with CEBV but not in those from patients with acute IM by Southern blot hybridization. Spontaneous lymphoblastoid cell lines could be easily established from PBMC of patients with acute IM but not from PBMC of patients with CEBV, despite several attempts. Viral capsid antigen and early antigens were not observed in cultured cells from patients with CEBV. These results suggest that the main site of EBV replication in CEBV might not be oropharyngeal epithelial cells. PMID- 2559903 TI - The non-effect of pirenzepine in dietary resistant irritable bowel syndrome. AB - No subjective or objective evidence of beneficial effect was observed from the use of Pirenzepine in irritable bowel syndrome (IBS) patients who had not responded to a high fibre diet. PMID- 2559905 TI - Imaging benzodiazepine receptors in the human brain by single photon emission computed tomography (SPECT). AB - Recently, [123I]Ro16-0154 has been introduced as a potential benzodiazepine (BZD) receptor imaging tracer for SPECT. We investigated the biodistribution of [123I]Ro16-0154 in humans and compared it with the animal data. There was a rapid and high 123I-uptake in the brain comparable to rCBF. In the following images, however, regionally different changes were observed, resulting in a subsequent intracerebral retention parallel to the known distribution of BZD receptors. The results suggest that [123I]Ro16-0154 is of potential clinical use as a BZD receptor imaging agent. PMID- 2559906 TI - Testicular dysfunction in the adjuvant-induced arthritic rat. AB - Adjuvant-induced arthritis, an autoimmune disease similar to rheumatoid arthritis, was used to investigate possible mechanisms of immune system modulation of the reproductive system. This laboratory previously reported that arthritic male rats have reduced serum testosterone and elevated serum LH concentrations. In the experiments described here, serum prolactin levels were not significantly different in arthritic animals compared with non-injected control animals. Neither reduced food consumption of arthritic rats nor the injection vehicle appear to cause a reduction of serum testosterone. Serum corticosterone was significantly elevated in the arthritic group compared with both the non-injected or the vehicle-injected control animals. Testicular cells from arthritic animals secrete significantly less testosterone in vitro compared with cells from non-injected control animals, both basally and in response to dbcAMP and hCG. In summary, the reduced serum testosterone of arthritic animals appears to be the result of a testicular dysfunction. PMID- 2559904 TI - [Condylomata acuminata gigantea with detection of HPV-6-DNA. A case report with adjuvant systemic IFN-gamma therapy]. AB - A case history of a 65-year-old man with giant condylomata (Buschke-Loewenstein tumour) of 30 years' duration is presented. The lesions were histologically confirmed, there were no signs of metastatic spread. HPV 6a DNA has been identified in Buschke-Loewenstein tumours and in different types of papilloma. After surgical removal in several sessions and treatment with gamma interferon by subcutaneous injection into the abdominal skin the patient has remained almost free of lesions. PMID- 2559907 TI - Stimulation of the proliferation and differentiation of Leydig cell precursors after the destruction of existing Leydig cells with ethane dimethyl sulphonate (EDS) can take place in the absence of LH. AB - In hypophysectomized rats, 2 days after the administration of the cytotoxic drug ethane dimethyl sulphonate (EDS), the proliferative activity of Leydig cell precursors increased six-fold. Thus, factors other than LH act locally to stimulate the proliferation of precursor cells after EDS. Twenty-six days after EDS administration, neither cells with the morphological characteristics of Leydig cells nor histochemical enzyme activities, such as 3 beta-HSD and alpha naphtyl esterase, could be detected in testis tissue. In hypophysectomized rats treated daily with hCG (100 iu) for 7 days, starting at 26 days after EDS, the number of Leydig cells was increased to 48 +/- 11 cells (per 1000 Sertoli cells), which is approximately 4.5% of the intact control level. 3 beta-HSD and alpha naphtyl esterase activity could be detected, and plasma testosterone levels had increased 15-fold compared with the hypophysectomized controls. These results show that proliferation and some differentiation of precursor cells along the Leydig cell lineage can occur independent of LH, but the final stages of the differentiation process require hCG stimulation. PMID- 2559909 TI - KS-504 compounds, novel inhibitors of Ca2+ and calmodulin-dependent cyclic nucleotide phosphodiesterase from Mollisia ventosa. AB - Novel inhibitors of Ca2+ and calmodulin-dependent cyclic nucleotide phosphodiesterase were isolated from the culture broth of a fungus, Mollisia ventosa KAC-1148. They were designated as KS-504a, KS-504b and KS-504d. A degradative product of KS-504a, called KS-504e, was also isolated and found to inhibit the enzyme. The 50% inhibitory concentration (IC50) values of KS-504a, KS 504b, KS-504d and KS-504e for bovine brain enzyme were 122, 109, greater than 500 and 139 microM, respectively. PMID- 2559908 TI - Development of transmembrane signaling in the fetal rat Leydig cell. AB - Gonadotropin binding to the adult Leydig cell activates a GTP binding protein that interacts with adenylate cyclase to increase cAMP production within the cell. The increased production of cAMP stimulates steroidogenesis and leads to an increase in testosterone production and secretion. The fetal Leydig cell responds to LH with an increase in cAMP and testosterone production as early as 15.5 days of gestation, although the specific mechanism of transmembrane signaling has not been characterized. Fetal rat testis cells from 13.5-20.5 days of gestation were treated with dibutyryl cAMP (dbcAMP), cholera toxin, and hCG to determine the onset of steroidogenesis stimulation by activation of each moiety in the transmembrane signaling system of the fetal Leydig cell. Maximal stimulation at each age from 14.5 through 20.5 days of gestation was achieved with 1 mM dbcAMP, 500 ng/ml cholera toxin, or 10 ng/ml hCG. At 13.5 days of gestation, fetal testes did not produce any testosterone. These findings indicate that a cholera toxin sensitive, stimulatory guanine-nucleotide regulatory protein is functional in the fetal Leydig cell as early as 14.5 days of gestation. The LH receptor becomes functional in the transmembrane signaling system of the fetal Leydig cell at 14.5 days of gestation. PMID- 2559910 TI - Induction by herbimycin A of contact inhibition in v-src-expressed cells. AB - Herbimycin A, an inhibitor of pp60src tyrosin kinase, caused src oncogene expressed cells to become sensitive to contact inhibition, but did not affect ras oncogene-expressed cells. The cell lines tested were temperature sensitive v-src- and temperature sensitive v-ras-integrated nontransformed rat kidney cell line (NRK) (srctsNRK and rastsNRK, respectively) and a wild-type v-src-integrated NIH3T3 (src3T3). srctsNRK cells in densely populated cultures (plated at 1.25 x 10(4) cells/cm2), grown at 33 degrees C in the presence of 0.45 micrograms/ml of herbimycin A, ceased the cell cycle at the G0-G1 stage within 2 days, and the cells showed normal morphology. Upon removal of herbimycin A, the quiescent cells resumed the cell cycle in concert with morphological alteration from 'normal' to 'transformed', and proceeded through the S and M stages successively in a synchronized manner. Cells in the late S stage, compared with those in other stages of the cell cycle, were more sensitive to the killing effect of 5 fluorodeoxyuridine. Such synchronism of the cell cycle was not observed with sparsely populated cultures (2.5 x 10(3) cells/cm2); the cells resumed their asynchronous growth after removal of herbimycin A, although their morphology returned to 'transformed' as in the experiment with the densely populated cultures. The induction by herbimycin A of contact inhibition in densely populated cultures was also observed with src3T3 (grown at 37 degrees C) but not with rastsNRK (grown at 33 degrees C). PMID- 2559911 TI - Anti-herpesvirus activity of carbocyclic oxetanocin G in vitro. AB - A series of new compounds, carbocyclic oxetanocins, have been synthesized and their anti-herpesvirus activity determined. Carbocyclic oxetanocin G (OXT-G) was most active against herpes simplex virus (HSV) and human cytomegalovirus (HCMV) among carbocyclic oxetanocins tested; the median effective concentrations (EC50) for HSV-1, -2, and HCMV were 0.23, 0.04 and 0.40 micrograms/ml, respectively. The EC50 value of carbocyclic OXT-G against HSV-2 was significantly lower than those of acyclovir, ganciclovir (DHPG) and OXT-G, while the value for HCMV was comparable to those of DHPG and OXT-G. Carbocyclic OXT-G showed much higher activity against TK+ HSV-2 than against a TK- mutant, suggesting that this compound is a good substrate for HSV-2-induced TK. The antiviral activity of the compound was only partially reversed even by the addition of 100-fold excess deoxyguanosine. The results suggest that the mode of action of carbocyclic OXT-G is different from that of OXT-G. PMID- 2559912 TI - Characterisation and molecular cloning of the novel macrolide-streptogramin B resistance determinant from Staphylococcus epidermidis. AB - A total of 110 staphylococcal isolates from human skin were found to express a novel type of erythromycin resistance. The bacteria were resistant to 14-membered ring macrolides (MIC 32-128 mg/l) but were sensitive to 16-membered ring macrolides and lincosamides. Resistance to type B streptogramins was inducible by erythromycin. A similar phenotype, designated MS resistance, was previously described in clinical isolates of coagulase-negative staphylococci from the USA. In the UK, MS resistance is widely distributed in coagulase-negative staphylococci but was not detected in 100 erythromycin resistant clinical isolates of Staphylococcus aureus. Tests for susceptibility to a further 16 antibiotics failed to reveal any other selectable marker associated with the MS phenotype. Plasmid pattern analysis of 48 MS isolates showed considerable variability between strains and no common locus for the resistance determinant. In one strain of S. epidermidis co-resistance to tetracycline, penicillin and erythromycin (MS) was associated with a 31.5 kb plasmid, pUL5050 which replicated and expressed all three resistances when transformed into S. aureus RN4220. The MS resistance determinant was localised to a 1.9 kb fragment which was cloned on to the high-copy-number vector, pSK265. A constitutive mutant of S. aureus RN4220 containing the 1.9 kb fragment remained sensitive to clindamycin. This observation, together with the concentration-dependent induction (optimum 5 mg/l of erythromycin) of virginiamycin S resistance suggests that the MS phenotype is not due to altered expression of MLS resistance determinants (erm genes) but probably occurs via a different mechanism. PMID- 2559913 TI - Effects of sub-inhibitory concentrations of antibiotics on surface expression of ferripyochelin-binding protein in Pseudomonas aeruginosa. AB - Growth of Pseudomonas aeruginosa in medium containing sub-inhibitory concentrations of tobramycin, tetracycline or chloramphenicol repressed surface expression of the ferripyochelin binding protein (FBP). Ciprofloxacin did not repress FBP surface expression but increased the amount of detectable FBP. Sub MICs of tetracycline, tobramycin and chloramphenicol also reduced ferripyochelin uptake by whole cells. An additional Mr = 19,000 protein was detected with monoclonal antibody in outer membranes of cultures grown in the presence of tetracycline and chloramphenicol. This protein is presumed to be a precursor form of FBP. No other major changes in LPS migration patterns or outer membrane protein profiles were observed in cultures grown in the presence of these antibiotics. These data suggest that exposure of P. aeruginosa to sublethal doses of tobramycin, tetracycline or chloramphenicol can alter the ability of these organisms to acquire iron. PMID- 2559914 TI - Pre-operative embolization of juvenile nasopharyngeal angiofibromas. AB - Pre-operative embolization of juvenile nasopharyngeal angiofibroma was performed in 15 patients. The lesion was supplied by the internal carotid arteries (8 cases), by the internal maxillary artery (15 cases), the accessory meningeal artery (10 cases) and the ascending pharyngeal artery (10 cases). Superselective embolization of the external carotid artery feeders was performed with Ivalon particles, without neurological complications. Good control of per-operative blood loss was noted in 13 out of 15 cases, 2 patients presenting severe per operative venous bleeding. Recurrence was noted in only one patient, which could be controlled by reembolization. PMID- 2559915 TI - Semilunar osteoblastic metastases in breast carcinoma. A new radiological sign. AB - Forty-seven cases of osteoblastic skeletal metastases were examined, of which 26 were from primary carcinoma of the breast. In two of these cases the metastases were of an unusual semicircular configuration. In each case the patient survived for at least four years after the discovery of metastases. This appearance may represent a better form of host response to metastatic disease. PMID- 2559916 TI - Unusual presentation of an endocrine pancreatic tumor. PMID- 2559917 TI - Cystosarcoma phyllodes. PMID- 2559918 TI - Preadsorption of polymers on glass and silica to reduce fibrinogen adsorption. AB - Glass and silica beads were precoated with various polymers to obtain steric exclusion chromatography (SEC) supports which are nonadsorbant for hydrophilic macromolecules. The efficiency of this treatment was estimated by subsequent radiolabeled fibrinogen adsorption. The result obtained with a block copolymer was better than with various hydrophilic homopolymers. This ABA type block copolymer, where A is a poly(N-acetylethyleneimine) (PAEI) sequence and B a polyethylene oxide (PEO) sequence was preadsorbed at pH 4.5 and 25 degrees C; the fibrinogen adsorption was reduced to less than 5% of the value observed on untreated solid surfaces. Thus the hemocompatibility of solid supports should be increased by precoating with this block copolymer. Results for nonporous glass beads and porous silica particles were in good correlation. PMID- 2559919 TI - Synthesis and adsorption of a poly(N-acetylethyleneimine)-polyethyleneoxide-poly (N-acetylethyleneimine) triblock-copolymer at a silica/solution interface. Influence of its preadsorption on platelet adhesion and fibrinogen adsorption. AB - The synthesis of a triblock copolymer poly-(N-acetylethyleneimine) polyethylenoxide-poly(N-acet ylethyleneimine) includes two successive steps: the first is the functionalization of a poly(ethyleneglycol) precursor by creating sulfonic esters at its chain ends, the second uses these esters to initiate the cationic polymerization of 2-methyl-2-oxazoline. Homopolymers appear in the raw product; hence successive selective extractions of the copolymer with benzene and dioxane are necessary. The final yield in pure copolymer was 11%. The copolymer was characterized by UV and 1H-NMR spectrometry and light scattering. Adsorption isotherms were determined on silica, for varying pH and salt concentration. Optimum conditions for coating silica with the polymer were determined. The efficiency of this precoating to reduce the adsorption of fibrinogen was very high (99.2% reduction with respect to bare silica). Steric exclusion chromatography of a variety of proteins gave a satisfactory calibration curve. Platelet accumulation on copolymer precoated glass was reduced to 10-20% of its value on bare glass, a result superior to that obtained by albumin passivation of the same glass surface. PMID- 2559920 TI - Embryonic chicken fibroblast collagen binding proteins: distribution, role in substratum adhesion, and relationship to integrins. AB - Collagen binding proteins (CBP) are hydrophobic, cell surface polypeptides, isolated by collagen affinity chromatography. Antibodies to CBPs inhibit the attachment of embryonic chicken heart fibroblasts to native type I collagen fibrils in a dose-dependent manner. The CBP antibodies also induce rounding and detachment of cells adherent to a planar substratum. This process of antibody mediated substratum detachment resulted in a clustering of CBP and cell associated extracellular matrix at the cell surface, and the rearrangement of filamentous actin. Other functional studies showed that cells grown within a three-dimensional gel of type I collagen cannot be immunostained at the cell surface with CBP antibodies. However, treatment of cultures with purified collagenase, unmasks immunoreactive sites and permits strong cell surface immunolabeling. This result suggests that collagen sterically blocks antibody access to CBP. Finally, we show that antibodies to CBP recognize purified avian integrin beta subunits; and that antibodies to avian integrins recognize a 100,000 Mr CBP. These data demonstrate that chicken embryonic fibroblasts possess surface polypeptides that mediate adhesion to type I collagen, and suggest that two of these proteins are related to the integrin family. PMID- 2559921 TI - Detection of immediate early, early and late antigens of human cytomegalovirus by flow cytometry. AB - The development of selective inhibitors of human cytomegalovirus (CMV) should lead to a better understanding of the mode of replication of CMV. A flow cytometric method was developed to monitor the expression of CMV antigens in CMV infected human embryonic lung fibroblasts. The procedure is based on an indirect immunofluorescence assay using monoclonal antibodies directed against CMV specific (immediate early, early, late) antigens and goat anti-murine IgG labeled with fluorescein isothiocyanate. Flow cytometric analysis clearly distinguished between the uninfected and infected cell population. There was a time-dependent appearance of CMV-specific antigens and a close correlation between the multiplicity of infection and the ratio of infected to uninfected cells. The method allows an accurate determination of the percentage of CMV-infected cells in the whole cell population and of the time of appearance of the viral antigens. PMID- 2559922 TI - The use of a hybridization assay for the study of host defences against herpes simplex virus. AB - A rapid and simple hybridization assay was developed as an alternative for virus titration for the investigation of host resistance against HSV-1 infections in vitro. The probe which was constructed for this assay was shown to be HSV-1 specific. When a monolayer of fibroblasts was infected for 24 h before hybridization, 15 PFU were detected reliably. A plateau in hybridization levels was found when the multiplicity of infection reached 1. In order to demonstrate the applicability of the probe for the study of host defences against HSV in vitro, fibroblasts were infected with HSV in the presence of different numbers of adherent cells and different concentrations of serum containing high titres of anti-HSV antibodies and complement. After 20 h of incubation, samples were lysed, spotted on Zetaprobe filter paper and hybridized with a 32P-labelled RNA probe. Spots were counted for radioactivity. The radioactivity was taken as a measure of the success of infection. Results showed that at high (10%) concentrations of serum containing high titres of anti-HSV antibodies and complement neutralization plays an important role. At low (1%) concentrations of serum containing high titres of anti-HSV antibodies and complement the phagocytic role of adherent cells becomes the dominant factor in preventing infection of the fibroblasts. However, when the number of infectious particles is increased, the protection provided by adherent cells is overwhelmed. PMID- 2559923 TI - Detection of human papilloma virus (HPV) DNA in genital biopsy specimens by in situ hybridization with digoxigenin-labeled probes. AB - The presence of human papillomavirus (HPV) nucleotide sequences in paraffin sections of genital biopsies was examined by in situ hybridization using non isotopic, digoxigenin-labeled probes representing HPV types 11, 16 and 18. Digoxigenin-labeling of the probes was performed using DNA labeling and a commercially provided detection kit. Hybridization was performed under stringent conditions. The hybrids were detected by using anti-digoxigenin alkaline phosphatase conjugate and visualized with enzyme catalyzed color reaction. In situ hybridization with digoxigenin-labeled probes was a useful technique for identification of HPV infection. The results were compared with the results obtained with radiolabeled DNA probes. The sensitivity of the digoxigenin-labeled probes was equal to the sensitivity of the radiolabeled probes. The background with digoxigenin-labeled probes was very low. Using nonradioactive probes the localization of hybrids at the cellular level was better than 35S-labeled probes. PMID- 2559924 TI - Rapid detection of cytomegalovirus in bronchoalveolar lavage specimens from marrow transplant patients: evaluation of a direct fluorescein-conjugated monoclonal antibody reagent. AB - An FITC-conjugated monoclonal antibody reagent containing three CMV-specific monoclonal antibodies was evaluated for the rapid detection of CMV in bronchoalveolar lavage (BAL) cytospin preparations by direct IF (DFA). Eighty-six BAL samples from 72 marrow transplant patients were inoculated into both centrifugation and standard cell culture. CMV was detected in 49/86 (57%) BAL samples. DFA detected 37/46 (80%) samples which were positive in centrifugation culture. While DFA staining lacked the sensitivity (overall sensitivity 38/49, 78%) to replace either standard or centrifugation culture, the total laboratory time needed to complete the DFA was only 1.5 h and its concurrent use with centrifugation culture can provide rapid specific diagnosis of CMV pneumonia. PMID- 2559925 TI - An indirect immunofluorescence method for detection of infectious BK virus in urine. AB - An indirect immunofluorescence (IF) method is described for the detection of infectious BK virus in urine within seven days in contrast to up to three months or longer using routine tissue culture. Virus is pelleted from the urine, inoculated onto pre-formed monolayers of human embryo lung (HEL) fibroblasts, and infected cells are detected in an indirect fluorescent antibody test using a human serum. The sensitivity of the IF method is 91%. Positive isolates may be confirmed as BK virus using specific rabbit antisera on the original inoculated cultures. Furthermore, a virus stock may be grown up by passage from the original culture fluids for further studies such as DNA analysis. As a broadly-reactive human serum is used for screening the cultures, other viruses which grow in HEL cells, such as CMV, may also be detected. PMID- 2559927 TI - Selenium addition to fertilizers effectively increased the serum levels of this element in the Finnish population. PMID- 2559926 TI - Synthetic macrophages: antigen presentation by liposomes bearing class II major histocompatibility complex (MHC) and membrane interleukin-1 (IL-1). PMID- 2559928 TI - [Fibrous histiocytoma of the orbit]. AB - The authors reported a case, a large orbital fibrous histiocytoma, in a 14 year old girl. They emphasize nosologic problems and histologic aspects of this benign tumor. Because of the localisation and the size of the tumor, a double anterior orbitotomy (superior and inferior) was necessary for complete renoval. Anatomic, functional and esthetic results were satisfactory. PMID- 2559929 TI - Serotonin modulated Ca++ dependent K+ channels in alloimmune effector cell lytic function. AB - Potassium channel activity has been implicated in the lytic function of cloned murine effector T lymphocytes (5) and human NK cells (12) as well as in the initiation of the injury process in tumor cells (23). In the present studies, the effects of various K+ channel blockers on the cytolytic function of in vivo derived alloimmune lymphocytes towards P815 tumor cells were evaluated. The classical K+ channel blocker 4-aminopyridine (4-AP), the naturally occurring monoamine serotonin (5-hydroxytryptamine, 5-HT) and its agonist, quipazine, as well as the Ca++ dependent K+ channel blocker quinidine were chosen for investigation based on their known ion channel gating properties. These agents, when present in the assay medium, inhibited in a dose dependent manner the lysis of P815 tumor cells as measured by specific 51Cr release. Preincubation of effector lymphocytes with the various K+ channel blockers resulted in greater inhibition of lysis than did the preincubation of target cells. The 5-HT agonist quipazine was of particular interest in that it inhibited the lytic process with equal effectiveness when continuously present in the assay medium or when the effector cells alone were preincubated. Quinidine was used to investigate whether Ca++ dependent K+ channels were the predominant ion channel involved in the lytic process. When present during the lytic assay, quinidine was similar to quipazine in terms of their dose range at which they inhibited the lytic process. These results indicate that 5-HT sensitive Ca++ dependent K+ channels are likely to be involved in the delivery of lytic signal(s) by immune effector lymphocytes and suggests that neuroendocrine products may modulate the functional activity of in vivo derived lymphocytes. PMID- 2559930 TI - Sensitive detection of two IgG Fc receptors of mouse macrophages by chemiluminescence analysis. AB - Luminol-enhanced chemiluminescence assay was used to detect the surface expression and the consequent activation of receptors (FcRI and FcRII) of murine macrophages (M phi s). When murine IgG2a was used for the specific detection of FcRI and IgG2b for FcRII, a newly established procedure enabled us to detect the activation of each receptor with as few as 3 X 10(5) M phi s. Briefly, TNP-SRBC coated with monoclonal IgG2a or IgG2b antibodies directed to TNP (sensitized SRBC) were used as reagent, in the presence of 1 X 10(-5) M luminol, and the emission was measured with a liquid scintillation counter. When results obtained by chemiluminescence counting were compared to the results obtained by the rosette formation by adding the same SRBC reagent to peritoneal M phi s obtained after ip injection of Listeria, fortified chemiluminescence counting allowed us to obtain a more definite answer about the activation of each receptor. Under the conditions established, the specific activation of FcRI was obtained by the addition of rIFN alpha A/D to the resident M phi s in vitro and the specific activation of spleen M phi FcRII by iv injection of IAP (Immunosuppressive acidic protein) into mice. These two results supported the independence of the two receptors detected by the assay. PMID- 2559931 TI - Inhibition of natural killer activity by calcitonin gene-related peptide. AB - The effect of calcitonin gene-related peptide (CGRP) on natural killer (NK) cell activity in spleen cells from Balb/c mice and nude mice was studied. CGRP dose dependently (10(-9) to 10(-7) M) inhibited NK activity of spleen cells from both strains of mice. This inhibitory effect was observed at the effector to target ratios of 12.5:1 to 100:1. Maximum inhibition by 10(-7) M CGRP was about 60%. The inhibition of NK activity by CGRP was also observed in anti-Thy 1.2 plus complement treated Balb/c spleen cells. Furthermore, when cells were treated with 10(-9) to 10(-7) M CGRP the concentration of intracellular cyclic AMP increased in spleen cells of nude mice. The characteristics of these cells were similar to those of NK cells, (1) being petri dish and nylon wool nonadherent, (2) expressing asialo GM1 antigen, and (3) lacking readily detectable Thy 1 antigen and immunoglobulin. In addition, the intravenous injection of asialo GM1 completely abolished NK activity in spleen cells from nude mice and the increase in intracellular cyclic AMP in spleen cells by CGRP was less in spleen cells from mice given an anti-asialo GM1 injection. Our present study suggests that CGRP inhibits NK cell activity by increasing the intracellular cyclic AMP concentration. CGRP may be implicated in the regulation of NK function. PMID- 2559932 TI - [Congenital mesoblastic nephroma (Bolande's tumor). Prenatal diagnosis]. AB - The authors report a new case of the antenatal ultrasound diagnosis of a congenital mesoblastic nephroma (also called Bolande's tumor) at 33 weeks of amenorrhea. This tumor was found to be associated with acute hydramnios involving premature labor and fetal death by respiratory embarrassment. Congenital mesoblastic nephroma is a very rare entity but is the predominant renal neoplasm in the immediate neonate period. The differential diagnosis between Bolande's tumor and nephroblastoma (Wilm's tumor) is now clear since histopathological features have been precisely described by Bolande. Clinical and pathological features of the renal disease are discussed. The prognosis is excellent for most cases requiring only surgical treatment (nephrectomy). PMID- 2559933 TI - Generation and characterization of a human monoclonal antibody against phenolic glycolipid-I of M. leprae. AB - The development of an Epstein-Barr virus transformed human B-cell line secreting a monoclonal antibody (MoAb), KR2/B5 is described. KR2/B5 is an IgM type of antibody and is highly specific for phenolic glycolipid-I (PGL-I) a component unique to M. leprae. The MoAb appears to be directed against the terminal sugar residue of the immunodominant trisaccharide component of PGL-I. PMID- 2559934 TI - Mandibulotomy: a surgical approach to oral and pharyngeal lesions. AB - The history of mandibulotomy is reviewed. The surgical steps involved and the variations of techniques are discussed. Two illustrative cases are reported and the clinical applications of the approach are discussed. It is concluded that mandibulotomy has a place in the management of malignancies as well as awkwardly situated benign lesions as well as malignancies. PMID- 2559935 TI - [Rotavirus infections in traveler's diarrhea]. AB - We tried to isolate rotaviruses from travelers with diarrhea arriving at Nagoya International Airport. 1. Ten cases revealed positive for rotavirus out of 334 diarrheal patients tested during a period from 1985 to 1988. 2. Most of the rotavirus positive cases were in their forties or in their fifties. 3. Geographic distribution of the infected area of the cases was not concentrated with specific countries. 4. Diarrhea was the most frequent clinical manifestations of the cases. 5. There was no cluster in seasonal distribution of the cases. PMID- 2559936 TI - [Na+,K(+)-ATPase: genes, expression and membrane insertion]. PMID- 2559937 TI - [Immortalization of human dental pulp cells with transfecting of the plasmid, pMT1-neo]. AB - A human dental pulp tissue was explanted in culture and the outgrowing cells were transfected with the plasmid, pMT1-neo, which included the early region of SV 40 DNA and the neomycin-resistant gene. The transfected cells were cloned and cultured beyond the period of senescence for the non-transfected HOP cells, over 170 passages (360 population doubling levels and 730 days). The characteristics of the transfected LSC cells and the non-transfected HOP cells were investigated during in vitro aging. The results were as follows: 1. The alkaline phosphatase (ALPase) activity of the HOP cells decreased as the culture passages increased. In contrast, the ALPase activity of the LSC cells did not change during the entire serial culture period. 2. The ALPase inhibitory test indicated that the ALPase in the LSC cells was the bone/liver/kidney type. 3. The addition of 250 micrograms/ml of L-ascorbic acid resulted in an increased collagen synthesis compared with that of 50 micrograms/ml. 4. The growth rate and the ALPase activity of the LSC cells were affected by 1 alpha, 25-dihydroxyvitamin D3, L ascorbic acid, beta-sodium glycerophosphate, epidermal growth factor or transforming growth factor-beta even after the serial culture. These results suggest that the LSC cells preserve some properties of the dental pulp and may be useful in the future research for exploring the mechanisms of the dental hard tissue formation and for testing the biocompatibility of the dental restorative materials. PMID- 2559939 TI - Pancreatic vasoactive intestinal polypeptide-secreting tumor: report of a case. AB - A 58-year-old Taiwanese woman was admitted to Mackay Memorial Hospital for evaluation profuse watery diarrhea. She presented with watery diarrhea, hypokalemia, and hypochlorhydria, and had experienced these problems for 2 years prior to admission. A pancreatic tumor with liver metastasis was noted by the ultrasound, abdominal CT scanning, and angiography studies. Surgical exploration disclosed an ill-defined ovoid tumor in the body and tail of the pancreas measuring 8 x 3 x 3 cm. The immunohistochemistry study of the tumor for VIP immunoreactivity (VIP: vasoactive intestinal polypeptide) was markedly positive, and also stained slightly positive for other peptides, including pancreatic polypeptide (PP), calcitonin, glucagon, and neuron-specific enolase (NSE). Postoperatively, the patient recovered immediately from her symptoms and there has been no evidence of recurrence during the past 8 months of follow-up. PMID- 2559938 TI - A simple culture method of fat cells from mature fat tissue fragments. AB - To obtain immature fat cells in vitro, we used a primary culture of undigested mature fat tissue fragments. The immature fat cells, i.e., fibroblast-like fat cells, proliferated extensively from the fat tissue and differentiated after reaching confluence. The process of differentiation was assumed by the development of intracytoplasmic lipid droplets and by the triglyceride content in the cells. Cellular differentiation was induced in high percentages (over 70-80%) of the cells in the medium containing high glucose concentrations (200 mg/dl) supplemented with 10-20% newborn calf serum. The intracellular accumulation of triglyceride was also enhanced by insulin administration. In these cells, a reciprocal relationship was observed between proliferation and differentiation. Fibroblast-like fat cells derived from mature fat tissue in this simple culture system are suitable for the study of the proliferation and differentiation of immature fat cells. PMID- 2559940 TI - Location and characterization of two functions on RP1 that inhibit the fertility of the IncW plasmid R388. AB - Two fertility-inhibition functions which reduce R388 (IncW) transfer were detected on RP1 (60 kb, IncP). The respective genes, fiwA and fiwB, were mapped by transposon insertion mutagenesis to the regions between coordinates 32.8 to 31.7 kb (fiwA), and 59.8 to 0.8 kb (fiwB). The fiwA function occurs in a non essential region of RP1 whereas fiwB is straddled by essential plasmid maintenance and host-range determinants and apparently coincides (or overlaps) with the gene for tellurite-resistance. PMID- 2559941 TI - Molecular analysis of a plasmid-encoded phenol hydroxylase from Pseudomonas CF600. AB - Pseudomonas strain CF600 is able to utilize phenol and 3,4-dimethylphenol as sole carbon and energy source. We demonstrate that growth on these substrates is by virtue of plasmid-encoded phenol hydroxylase and a meta-cleavage pathway. Screening of a genomic bank, with DNA from the previously cloned catechol 2,3 dioxygenase gene of the TOL plasmid pWW0, was used in the identification of a clone which could complement a phenol-hydroxylase-deficient transposon insertion mutant. Deletion mapping and polypeptide production analysis identified a 1.2 kb region of DNA encoding a 39.5 kDa polypeptide which mediated this complementation. Enzyme activities and growth properties of Pseudomonas strains harbouring this fragment on a broad-host-range expression vector indicate that phenol hydroxylase is a multicomponent enzyme containing the 39.5 kDa polypeptide as one component. PMID- 2559942 TI - The development of Chlamydia trachomatis inclusions within the host eukaryotic cell during interphase and mitosis. AB - The dynamic nature of Chlamydia trachomatis inclusions was studied by video and 35 mm time-lapse photomicrography of live cells, and by immunolocalization of inclusions in fixed cells. A serotype E isolate was used to infect the MCCoy cell line and endometrial epithelia. Then resulting inclusions were observed over 4 d. They appeared as slowly expanding fluid-filled membrane vesicles whose growth varied considerably, and which were subject to great physical distortion by the host cell during interphase and mitosis. When this distortion became extreme the inclusion was observed to divide. However, as inclusions were mobile within the cytoplasm and thus able to come into contact with each other, there was a net tendency for the opposite process of inclusion fusion to occur when cells contained more than one inclusion. The proportion of infected cells decreased with time as a result of host cell proliferation, despite transmission of inclusions to progeny at the time of mitosis. Inclusion growth physically disrupted karyokinesis and cytokinesis so that host cell division became distorted or blocked on the second or third day of infection. Cell death eventually occurred by a very rapid lysis event. PMID- 2559943 TI - Role of erythrosine in the inhibition of adhesion of Lactobacillus fermentum strain 737 to mouse stomach tissue. AB - The mechanism by which the food colour erythrosine inhibits the adhesion of Lactobacillus sp. to squamous epithelium in the mouse stomach was investigated using an in vitro adhesion assay. Inhibition of adhesion occurred only after growth of L. fermentum in erythrosine which bound to the bacterial cell surface. Erythrosine did not interfere with the receptor on the epithelial cell surface. Growth, but not the ATP content per cell, was affected by the presence of erythrosine in the growth medium. No consistent correlation between hydrophobicity and growth in two different broths was noted when erythrosine was present. Analyses of phenol/water extracts and transmission electron micrographs revealed no reduction in extracellular polysaccharide after growth in the presence of erythrosine. It was concluded that erythrosine affects bacterial metabolism thereby preventing production of the bacterial adhesin which is not the extracellular polysaccharide. PMID- 2559944 TI - Protein-mediated adhesion of Lactobacillus fermentum strain 737 to mouse stomach squamous epithelium. AB - The mechanism of adhesion of Lactobacillus fermentum strain 737 to mouse stomach squamous epithelium was investigated. Adhesion inhibition tests involving chelators, monosaccharides, periodate and concanavalin A and the use of bacteria grown in the presence of tunicamycin failed to clarify the adhesive mechanism. Washed bacterial cells had reduced adhesive capacity, except in the presence of spent broth culture supernatant fraction or cell washings. Spent culture supernatant fractions of erythrosine-supplemented broth did not enhance adhesion of washed cells. The adhesion-promoting factor(s) in the spent broth culture supernatant fractions and cell washings bound to both bacterial and epithelial cell surfaces, but did not promote adhesion of two other Lactobacillus strains which were not of mouse origin, thereby indicating host specificity for the adhesion-promoting activity. Chemical characteristics of the adhesion-promoting factor were determined by pretreatment of the dialysis retentate of spent broth culture supernatant fractions with proteolytic enzymes, concanavalin A-Sepharose or periodate before the adhesion assay. The adhesin was non-dialysable, pronase sensitive, heat sensitive at 100 degrees C, had no affinity for concanavalin A Sepharose and contained no carbohydrate groups active in the adhesion process. The protein profiles of dialysis retentates of spent broth culture supernatant fractions after bacterial growth in the absence and presence of erythrosine were determined by 2-dimensional SDS-PAGE. Gel filtration by HPLC was used for purification of an adhesion-promoting fraction. The host-specific adhesion of L. fermentum strain 737 was mediated by a protein, with an Mr of 12-13000, that was not detectable in cells grown in the presence of erythrosine. A model for the mode of binding of the adhesin to host epithelia and bacterial surfaces is proposed. PMID- 2559945 TI - Role of myeloperoxidase in the killing of Staphylococcus aureus by human neutrophils: studies with the myeloperoxidase inhibitor salicylhydroxamic acid. AB - We have used salicylhydroxamic acid (SHAM) to inhibit intraphagosomal myeloperoxidase activity in order to evaluate the role of this enzyme in the killing of Staphylococcus aureus by human neutrophils. 50 microM-SHAM reduced the luminol-dependent chemiluminescence response stimulated during phagocytosis of unopsonized latex beads and opsonized S. aureus by over 80% and 60%, respectively. When opsonized S. aureus were incubated with neutrophils, 45% were killed within 15 min incubation and 60% by 1 h. However, in neutrophil suspensions incubated with 50 microM-SHAM, only 13% were killed by 15 min whilst 71% still remained viable after 1 h. This inhibitor had no effect upon the number of bacteria phagocytosed or upon degranulation. In a cell-free system, 2.5 microM H2O2 alone killed 55% of the bacteria, whereas in the presence of myeloperoxidase (i.e. 10 mU myeloperoxidase and 2.5 microM-H2O2) virtually all of the bacteria were killed: the addition of 50 microM-SHAM abolished this myeloperoxidase enhanced killing but did not affect the H2O2-dependent killing. We therefore conclude that in normal neutrophils whilst H2O2 is required for killing of this pathogen, both myeloperoxidase-dependent and -independent pathways exist. PMID- 2559946 TI - The hexokinase isoenzyme PII of Saccharomyces cerevisiae ia a protein kinase. AB - The HXK2 gene product has an important role in controlling carbon catabolite repression in Saccharomyces cerevisiae. We have raised specific antibodies against the hexokinase PII protein and have demonstrated that it is a 58 kDa phosphoprotein with protein kinase activity. The predicted amino acid sequence of the HXK2 gene product has significant homology to the conserved catalytic domain of mammalian and yeast protein kinases. Protein kinase activity was located in a different domain of the protein from the hexose-phosphorylating activity. The hexokinase PII protein level remained unchanged in P2T22D mutant cells (hxk1 HXK2 glk1) growing in a complex medium with glucose. The protein kinase activity of hexokinase PII is regulated by the glucose concentration of the culture medium. Exit from the carbon catabolite repression phase and entry into derepression phase may be controlled, in part, by modulation of the 58 kDa protein kinase activity by changes in cyclic AMP concentration. PMID- 2559948 TI - Role of guanosine kinase in the utilization of guanosine for nucleotide synthesis in Escherichia coli. AB - Using purine auxotrophic strains of Escherichia coli with additional genetic lesions in the pathways of interconversion and salvage of purine compounds, we demonstrated the in vivo function of guanosine kinase and inosine kinase. Mutants with increased ability to utilize guanosine were isolated by plating cells on medium with guanosine as the sole purine source. These mutants had altered guanosine kinase activity and the mutations were mapped in the gene encoding guanosine kinase, gsk. Some of the mutants had acquired an additional genetic lesion in the purine de novo biosynthetic pathway, namely a purF, a purL or a purM mutation. A revised map location of the gsk gene is presented and the gene order established as proC-acrA-apt-adk-gsk-purE. PMID- 2559947 TI - Anaerobic growth of Escherichia coli on glycerol by importing genes of the dha regulon from Klebsiella pneumoniae. AB - The dha regulon of Klebsiella pneumoniae specifying fermentative dissimilation of glycerol was mobilized by the broad-host-range plasmid RP4:mini Mu and introduced conjugatively into Escherichia coli. The recipient E. coli was enabled to grow anaerobically on glycerol without added hydrogen acceptors, although its cell yield was less than that of K. pneumoniae. The reduced cell yield was probably due to the lack of the coenzyme-B12-dependent glycerol dehydratase of the dha system. This enzyme initiates the first step in an auxiliary pathway for disposal of the extra reducing equivalents from glycerol. The lack of this enzyme would also account for the absence of 1,3-propanediol (a hallmark fermentation product of glycerol) in the spent culture medium. In a control experiment, a large quantity of this compound was detected in a similar culture medium following the growth of K. pneumoniae. The other three known enzymes of the dha system, glycerol dehydrogenase, dihydroxyacetone kinase and 1,3-propanediol oxidoreductase, however, were synthesized at levels comparable to those found in K. pneumoniae. Regulation of the dha system in E. coli appeared to follow the same pattern as in K. pneumoniae: the three acquired enzymes were induced by glycerol, catabolite repressed by glucose, and glycerol dehydrogenase was post translationally inactivated during the shift from anaerobic to aerobic growth. The means by which the E. coli recipient can achieve redox balance without formation of 1,3-propanediol during anaerobic growth on glycerol remains to be discovered. PMID- 2559949 TI - Direct detection of human cytomegalovirus in urine specimens from renal transplant patients following polymerase chain reaction amplification. AB - A polymerase chain reaction (PCR) assay was used to amplify human cytomegalovirus (HCMV) directly from urine specimens taken from renal transplant patients. In serial urine samples from patients who had at least one specimen positive for HCMV; the PCR assay consistently detected the presence of HCMV DNA sequences, whereas virus detection by other tests such as enzyme-linked immunosorbent assay (ELISA), nonradioactive DNA hybridization assay, and virus isolation were variable. Of 37 specimens positive by PCR, 36 were positive by either ELISA, hybridization assay, or virus isolation. Infectious virus was detected in 13 of the 37 PCR-positive urines. HCMV DNA was detected by PCR in all samples that were positive for HCMV by either hybridization assay or virus isolation. The viral genome copy number was determined by PCR assay for several urine samples that were positive by virus isolation but negative for HCMV by ELISA or hybridization assay. Viral genome copy number estimates indicated the presence of HCMV at very low levels in these urines verifying the fidelity of the virus isolation procedures. The consistency of the PCR assay makes it an ideal method for detection of infection and monitoring antiviral drug therapy in patients infected with HCMV. PMID- 2559950 TI - Coronavirus-like particles in adults in Melbourne, Australia. AB - Coronavirus-like particle(s) (CVLP) are faecal-derived pleomorphic membrane bound virus-like particles characterised by a fringe of club-shaped spikes that measure about 27 nm in length. The association of CVLP with a variety of social, clinical, and epidemiological factors was examined after a 69 month survey of faeces received for routine testing at an infectious diseases hospital. CVLP was found most commonly in three groups: first, intellectually retarded individuals who were usually inmates of institutions; second, recent overseas travellers who were either Indochinese refugees/immigrants or were overseas travellers who had usually visited developing communities for lengthy periods; and, third, male homosexuals who had a history of multiple sexual contacts and/or venereal disease. It was concluded that the excretion of CVLP had a strong association with unhygienic living or working conditions irrespective of any clinical symptoms the individual might show. PMID- 2559951 TI - Effect of 3'-azido-3'-deoxythymidine on replication of duck hepatitis B virus in vivo and in vitro. AB - 3'-Azido-3'-deoxythymidine (AZT) inhibits the replication of the human immunodeficiency virus (HIV) by blocking the formation of the phosphodiester bond and has been used clinically for the treatment of HIV infection. To assess the effect of AZT on the replication of hepadnaviruses, which replicate through reverse transcription, both the liver tissue and primary cultured hepatocytes from ducklings previously infected with duck hepatitis B virus (DHBV) were examined for DHBV DNA before and after the treatment with AZT. We did not observe suppression of DHBV replication at any doses in our system as measured by viral DNA synthesis in infected duck hepatocytes. The data strongly suggest that AZT has no inhibitory effect on DHBV reverse transcriptase. PMID- 2559952 TI - Pig erythrocyte ghost cells used for concentration of enteric viruses from experimentally contaminated clinical specimens. AB - A simple, rapid and efficient procedure of virus concentration from urine, cerebrospinal fluid (CSF), and feces was developed. Pig erythrocyte ghost cells were used to adsorb and elute such viruses as poliovirus LSC-1, echovirus 6, and human rotavirus (clinical isolate). In urine and CSF, the adsorption efficiency range was 80-100% and elution was from 85% to greater than 100%. In addition, poliovirus LSC-1 was used as an experimental model to examine this procedure under various clinical conditions, such as calcium, glucose, amino acids, and urea at high concentrations. These were added to normal urine specimens to simulate pathological conditions. The results suggest that pig erythrocyte ghost cells are an efficient matrix for adsorption and elution of enteric viruses found in clinical specimens of urine, CSF, and feces. This method might be useful in virus concentration from clinical specimens and for preparative microscopy and other clinical laboratory methods that require subsequent virus concentration. PMID- 2559953 TI - Evaluation of a new enzyme-linked immunosorbent assay (ELISA) in the diagnosis of rhinovirus infection. AB - This study describes the evaluation of a newly developed ELISA for the direct detection of rhinovirus antigens in nasal washings. Of 54 volunteers inoculated with 100 TCID50 of human rhinovirus type 2 (HRV-2), 50 (96.6%) and 32 (59%) excreted antigen and virus on at least 1 of 3 days investigated, respectively. Thirty-three (61%) had significant rises in rhinovirus-specific IgA by ELISA. Twelve (22%) developed symptoms of colds. Generally the ELISA detected antigen more frequently in volunteers later in the course of infection and provided evidence of infection in a higher proportion of asymptomatic compared with symptomatic volunteers. On the other hand, virus isolation detected virus more frequently earlier in the course of infection and in a higher proportion of symptomatic compared with asymptomatic volunteers. We conclude that rhinovirus antigen detection by ELISA is a simple, rapid, sensitive, and practical test to diagnose a rhinovirus infection and potentially a viable alternative to virus isolation. PMID- 2559954 TI - Detection by antibody probes of human papillomavirus type 6 E5 proteins in respiratory papillomata. AB - We have demonstrated the expression of proteins arising from the E5a and E5b open reading frames (ORFs) of human papillomavirus type 6c (HPV-6c) in respiratory tract papillomata. Recombinant plasmids were constructed to express the ORFs in the bacterial vectors pATH and pRIT2T. Fusion proteins were purified and injected into rabbits to produce polyclonal antibodies. Characterized antibodies generated against these fusion proteins were used in immunoperoxidase assays to identify the presence and distribution of HPV-6 E5 proteins in biopsy specimens of respiratory tract papillomata. The results showed that the E5a and E5b proteins were distributed throughout the thickness of the epithelium in the papillomata but not in the basal layer. The proteins were found in nuclei and in the cytoplasm of koilocytotic cells. Positive reactivity with a similar distribution in the epithelium and subcellular location was obtained in papillomata induced by other HPV-6 subtypes. This cross-reactivity was not unexpected, since nucleotide and amino acid (aa) sequence comparisons between HPV-6c and -6e demonstrated 79% sequence identity with 15 aa substitutions in the 91 aa of E5a. The E5b ORF of HPV-6c has the potential to encode a protein of 74 aa that differed at 28 positions compared with the 72 aa of HPV-6e. PMID- 2559955 TI - Improved sensitivity of detection by avidin-biotin complex (ABC) immunocytochemistry in Epstein-Barr virus serology. AB - Serum antibodies against Epstein-Barr virus (EBV)-determined antigens have traditionally been titrated by the indirect immunofluorescence (IIF) technique. The avidin-biotin complex (ABC) immunocytochemical technique was used to determine the serum levels of IgA against EBV viral capsid antigen (IgA/VCA) and IgA against EBV early antigen (IgA/EA) in sera of 106 nasopharyngeal carcinoma (NPC) patients prior to treatment and 100 normal individuals. The sensitivity of the ABC technique is enhanced by an amplification of the antigen-antibody reaction, which involves the binding of the enzyme-linked ABC to the second biotinylated antibody. There was a good correlation (r = 0.9988) between ABC and IIF-determined IgA/VCA-positive titres, with the ABC technique being more sensitive than IIF in the detection of IgA/VCA in NPC sera: 94% (99/106) and 76% (80/106), respectively. The frequency of IgA/EA reactivity in NPC sera was also markedly increased by immunodetection with the ABC technique as compared with IIF technique: 63% (69/106) and 28% (30/106) respectively. Both the immunocytochemical techniques were equally specific in discriminating between elevated serum titres of IgA/VCA and IgA/EA in NPC sera from normal human sera. PMID- 2559956 TI - Astrocytic reaction predominance in chronic encephalitis of Junin virus-infected rats. AB - Junin virus antigen distribution and astrocytic reaction to prolonged infection were characterized in rat brain by the PAP technique. During the acute stage of neurologic disease following intracerebral inoculation, Junin antigen was detected in 100% of animals, strongly in most neurons but also to a much lesser degree in scattered astrocytes, dropping to 20% of rats at 540 days postinfection. Initially labeled in all brain areas, viral antigen gradually disappeared from hippocampus but persisted irregularly in cerebral cortex, basal ganglia, Purkinje cells, pons, and medulla oblongata. Such a pattern suggests that specific neuronal subpopulations, in spite of apparently unaltered cell morphology, may persistently harbor the virus, leading on occasion to a delayed neurologic syndrome. During both the acute and chronic stages of disease, a mild inflammatory exudate was observed, characterized by the presence of T and B lymphocytes, as well as macrophages and unidentified round cells. GFAP immunostaining showed increased astrocytic reaction as infection lapsed into chronicity. Corpus callosum, hippocampus, and cerebellum exhibited the sharpest reactive astrocytosis, followed by basal ganglia, pons, and medulla oblongata, whereas in cerebral cortex it was considerably less. Astrocyte activation, which failed to correlate with viral antigen presence in neurons, seems to result from a generalized condition, possibly including diffusible brain factors triggered by viral infection. Such widespread astroglial reaction may thus contribute to the outcome of the late neurologic syndrome. PMID- 2559957 TI - Dietary guar gum effects on postprandial blood glucose, insulin and hydroxyproline in humans. AB - Meals (425 kcal) containing various doses of guar gum (0, 2.5, 7.5 or 12.5 g) were ingested by nine healthy male subjects after a 12-h fast. The rise in blood glucose was higher after the control meal without guar gum than after the guar gum-containing meals, which all gave a similar rise in glucose. In contrast, increased doses of guar gum led to a greater reduction in the postprandial rise in insulin. The postprandial increase in serum hydroxyproline, an amino acid added to all meals, was decreased in a similar manner by all of the guar gum doses. Gastric emptying was measured after the control meal without guar gum and the meal containing 12.5 g of guar gum by monitoring 51Cr, which was added to the meals. Guar gum was found to reduce the variation between individuals, as well as the initial rate of gastric emptying, which correlated with changes in both serum hydroxyproline (rs = 0.93, P less than 0.01) and blood glucose (rs = 0.83, P less than 0.01). The effectiveness of guar gum in reducing postprandial response was lost after heating and homogenization for canning. A threshold in the reduction in rise of glucose or hydroxyproline was reached with the lowest dose (2.5 g) of viscous guar gum; larger doses had no additional effects. The reduced absorption seems to be an effect of a slower gastric emptying rate. PMID- 2559958 TI - Rat bioassay of wheat bran folate and effects of intestinal bacteria. AB - This study estimates the folate endogenous to a food material (wheat bran) and examines the role of intestinal bacteria in the rat bioassay for folate. After a 4-wk folate depletion period, rats were fed for an additional 4 wk basal diets with or without 0.5% phthalylsulfacetamide and with 100, 200 or 300 g of wheat bran; or 50, 100 or 150 g of xylan; or 0, 0.25, 0.50 or 0.75 mg of folic acid added per kg of basal diet. Xylan increased both liver and fecal folate, and this effect was nearly eliminated by phthalylsulfacetamide. Wheat bran contributed 1.6 micrograms of available folate per g of wheat bran without phthalylsulfacetamide in an apparently valid slope-ratio analysis. With the addition of phthalylsulfacetamide, liver folate increased in rats fed wheat bran diets and decreased in rats fed folic acid diets. The slope-ratio analysis for wheat bran folate with phthalylsulfacetamide became invalid due to a lack of intersection. Phthalylsulfacetamide had no effect on fecal folate excretion from rats fed the wheat bran diets. Further studies are needed on a variety of foods with and without phthalylsulfacetamide to evaluate the effect and importance of intestinal folate synthesis in the rat. PMID- 2559959 TI - [Clinicopathological study of salivary gland adenocarcinomas]. AB - A total of 27 cases of salivary gland adenocarcinomas were studied from clinicopathological view point. Adenocarcinomas of the salivary gland were microscopically subclassified into 3 groups according to Luna's classification: Salivary duct carcinomas histologically resembled the ductal carcinoma of the breast, displayed nuclear atypia and had poorer prognosis than the other subclasses of salivary gland adenocarcinomas. Terminal duct carcinomas lacked in nuclear atypia and displayed a variety of growth patterns, including papillary, cribriform, tubular, and solid. Some terminal duct carcinomas showed prominent mucin-production. Epithelial-myoepithelial carcinomas had clear cytoplasms and exuberant glycogen. In addition to the clinicopathological study, nuclear areas of the tumor cells were measured in each of the 27 salivary gland adenocarcinomas, and mean nuclear area (MMA) and standard deviation (SD) were calculated. The group with more than 50 microns 2 of MNA had poorer prognosis than the group with 50 microns 2 or less of MNA, and the group with more than 13 microns 2 of SD had poorer prognosis than the group with 13 microns 2 or less of SD. Finally, immunohistochemical study was performed against various markers including keratin, epithelial membrane antigen, lactoferrin, S-100 protein, CEA, etc., using the Avidin-biotin-peroxidase complex method. Lactoferrin was present in most of the salivary duct carcinomas, on the other hand, S-100 protein was detected in all of the five cases of the terminal duct carcinoma investigated. But immunohistochemical study is not especially useful in distinguishing subclasses of salivary gland adenocarcinomas or investigating the origin of tumor cells. PMID- 2559960 TI - [Detection of human papillomavirus genome in nasolaryngeal papillomas using digoxigenin labeled DNA probes]. AB - It is being reported that human papillomavirus (HPV) has been implicated in the pathogenesis of various neoplastic lesions of the genital organs. To investigate the etiological role of HPV and its types in nasolaryngeal papillomas, we retrospectively analyzed HPV genomes by nucleic acid hybridization methods; for detecting DNA and mRNA, we employed the recently developed nonradioactive (digoxigenin labeled) DNA probes and compared the results by radioisotope methods. In total, 43 cases of papillomatous lesions were examined. They were verruca vulgaris of the nasal vestibule (Nr = 2), nasal inverted papilloma (IP, Nr = 26), and laryngeal papilloma (Nr = 15). HPV types examined were type 2, 6, 11, 16 and 18. Two cases of verruca vulgaris were shown to contain HPV-2 DNA and its mRNA by in situ hybridization. HPV-11 DNA was detected in 3 cases (12%) of nasal inverted papilloma whereas HPV-16 was detected in 1 case (4%); the latter case was associated with squamous cell carcinoma. These results suggest that HPV may be implicated in the development of IP, and HPV-16 may play an important role in the malignant transformation of IP. In the cases of multiple laryngeal papilloma (Nr = 8, one juvenile type and 7 adult type), either HPV-6 or HPV-11 was detected at the high rate (6/8, 75%). The presence of the HPV genomes provides strong evidence for the HPV etiology of these laryngeal papillomas. Whereas in the cases of adult single laryngeal papilloma (Nr = 7), HPV was not detected. Technically, the sensitivity of digoxigenin (DIG) labeled DNA probe was almost same as 35S labeled probe by dot blot hybridization, thus we applied DIG labeled probe to Southern blot hybridization with low background. By in situ hybridization using digoxigenin labeled probes, the rates of HPV detection were almost equal to those by 35S labeled probes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2559961 TI - Optic nerve shadow enlargement in the Klippel-Trenaunay-Weber syndrome. AB - A 10-year-old girl with Klippel-Trenaunay-Weber syndrome (KTWS) showed neuroradiologic signs of enlarged optic nerve and medial rectus shadows. In 3 years of follow-up, her vision has remained normal. This is the first report of optic nerve enlargement in this syndrome. The case demonstrates that enlargement of the optic nerve shadows is not inevitably associated with visual deterioration. Optic nerve enlargement may be part of a hamartomatous process that involves other orbital contents. PMID- 2559962 TI - [Sensitivity of different cell lines to interferons: the relative antiviral activity as a function of the interferon subtype]. AB - The phenomenon that rHuIFN-alpha1(D) displays an apparently higher antiviral activity when assayed on bovine cells as compared to human cell lines was applied to the elucidation of the nature of recombinant HuIFN prepared in our institute. These investigations were carried out by using a microtitre test, which defines biological activity as the IFN concentration leading to 50% inhibition of the cytopathic effect of vesicular stomatitis virus (VSV). In addition, the ability of IFN to diminish the reproduction of infectious viruses was monitored. The two methods yielded similar results. With bovine cells, antiviral activities of the same order of magnitude were observed, regardless of the interferon types applied, i.e. rHuIFN-alpha 1, rHuIFN-alpha 2 and human leukocyte interferon. On human fibroblasts, however, rHuIFN-alpha 1 had an apparently 45 to 165 times lower activity than the other two interferons. On human WISH cells, the differences in apparent activity between the respective IFNs were even greater, with factors of up to 212 fold being observed. Still more distinctive were the effects on murine L 929 cells where an antiviral effect could be confirmed only for rHuIFN-alpha 1 whereas the other two interferons proved completely inactive. PMID- 2559963 TI - Biochemical aspects of the resistance to nourseothricin (streptothricin) of Escherichia coli strains. AB - In most cases Escherichia coli strains phenotypically resistant against nourseothricin (streptothricin) harbour a plasmid which codes for an acetyltransferase. This enzyme transfers an acetyl group from acetyl-coenzyme A to an amino group of the beta-lysine (peptide) chain of the antibiotic, thus inactivating it. Additionally, the penetrability for nourseothricin of the cell wall is drastically reduced in a high percentage of the resistant strains. Both resistance mechanisms seem to be independent of each other. PMID- 2559964 TI - Inhibition of 5-lipoxygenase by substituted 3,4-dihydro-2H-1,4-thiazines. AB - A series of substituted 3,4-dihydro-2H-1,4-thiazines inhibit 5-lipoxygenase from rat leukocytes and exhibit submicromolar IC50 values. A novel synthesis of these compounds was developed based on the formation of hydroxymethyleneamine 13 and its cyclization to the title compounds. The dihydrothiazines have low oxidation potentials, typically E1/2 is near 0.3 V, and a representative compound reduces Fe(III)(phen)3, with k = 10(5) M-1s-1. We propose that these lipophilic compounds bind to 5-lipoxygenase and reduce the iron in the active site, thus inactivating the enzyme. PMID- 2559965 TI - Sodium and potassium conductances in somatic membranes of rat Purkinje cells from organotypic cerebellar cultures. AB - 1. The somatic voltage-gated conductances of Purkinje cells in organotypic cultures (Gahwiler, 1981) were studied using the outside-out patch recording configuration of the patch-clamp technique (Hamill, Marty, Neher, Sakmann & Sigworth, 1981). 2. When activated by step depolarizations, the tetrodotoxin sensitive voltage-dependent Na+ current presented two distinct phases: an initial surge of inward current fluctuations which activates rapidly upon pulse onset and decays within 20-40 ms, and a later phase in which discrete bursts of single channel activity are interspersed with silent periods. 3. Ensemble fluctuation analysis of the current fluctuations during the early phase of the Na+ current and measurements of single channels during both early and late phases indicate that a single type of Na+ channel can account for both phases of the Na+ current. This channel has an elementary current amplitude of -2 pA at -40 mV. This amplitude did not vary significantly between -60 and -20 mV. The mean open time depended on membrane potential, increasing by a factor of three between -60 and 20 mV. 4. The early component of the Na+ current activated at a threshold of -60 mV and reached its maximum amplitude at -20, mid-point for the activation curve being -40 mV. Times-to-peak current decreased with membrane potential, from 3.5 ms at -60 mV to 0.3 ms at 0 mV. The decay phase of the current presented two exponential components, with time constants of 1.5 and 10 ms at -40 mV. The steady-state inactivation curve had a mid-point at -75 mV. 5. The late component of the Na+ current was observed in the voltage range from -60 to -20 mV, with a maximum at -40 mV. Its maximum amplitude corresponded to approximately 1.7% of the peak amplitude of the early component. 6. Macroscopic potassium currents were observed upon step depolarizations above a threshold of -30 mV. The currents activated in a voltage-dependent fashion, times-to-peak decreasing with depolarization, and partially inactivated during 40 ms depolarizing steps. Peak current amplitudes at any given membrane potential were decreased by depolarizing the holding potential. The macroscopic properties of the K+ current varied from patch to patch. 7. Two types of single-channel K+ currents were observed during steady-state depolarizations. The unitary current amplitudes were 2.7 and 10.4 pA at 30 mV, corresponding to chord conductances of 28 and 90 pS respectively.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2559966 TI - Interactive effects of isoprenaline, forskolin and acetylcholine on Ca2+ current in frog ventricular myocytes. AB - 1. Calcium currents (ICa) were measured in single cells isolated from frog ventricle using the whole-cell patch-clamp technique and a perfused pipette. The dose-dependent stimulatory effects of isoprenaline (Iso, 0.1-100 microM) and forskolin (Fo. 0.1-50 microM) on ICa were determined in the presence and absence of acetylcholine (ACh, 10 microM) and/or threshold concentrations of Fo (0.2 microM) and Iso (0.05 microM), respectively. EC50 (i.e. concentration of Iso or Fo at which the response was 50% of the maximum) and Emax (i.e. maximal stimulation of Ica expressed as percentage increase in ICa with respect to control) were measured under each condition. 2. ACh increased EC50 for the stimulatory action of Iso on ICa from 0.84 to 3.72 microM while it reduced Emax from 658 to 185%. Thus, ACh mainly reduced the efficacy of Iso to stimulate ICa. 3. ACh increased EC50 for the stimulatory action of Fo on ICa from 2.06 to 10.26 microM but only slightly reduced Emax from 893 to 778%. Thus, ACh mainly reduced the potency of Fo to stimulate ICa. 4. Intracellular perfusion with 100 microM of hydrolysis-resistant GTP analogues, GTP-gamma-S [guanosine-5'-O-(3 thiotriphosphate)] and Gpp (NH)p (5'-guanylylimido-diphosphate), had no effect on basal ICa but reduced by greater than 50% the stimulatory effect of 2 microM-Iso on ICa. 5. In the presence of Gpp(NH)p or GTP-gamma-S, Fo (3 microM) reversibly increased ICa by 490%, as compared to a 717% increase in control (GTP) intracellular solution. Although ACh could still inhibit Fo-stimulated ICa, the degree of inhibition was significantly smaller than in the presence of GTP. 6. Extracellular perfusion with low concentrations of a combination of Iso (33 nM) and Fo (330 nM) enhanced ICa to a much greater extent than did either agent alone at 3 times higher concentrations. Thus, low concentrations of Iso and Fo appear to increase ICa in a synergistic fashion. 7. ICa stimulated by a combination of Iso and Fo appeared to be more resistant to inhibition by ACh than when stimulated by either alone. It was the efficacy, rather than the potency, of ACh to inhibit ICa that was reduced upon dual stimulation of ICa. 8. In the presence of 0.2 microM-Fo, EC50 and Emax for the effects of Iso on ICa were 0.27 microM and 619%, respectively. By comparison with the effects of Iso alone, Fo reduced EC50 approximately 3 times with no significant change in maximal stimulation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2559968 TI - Second messenger-activated calcium influx in rat peritoneal mast cells. AB - 1. To study the regulation of calcium influx in non-excitable cells, membrane currents of rat peritoneal mast cells were recorded using the whole-cell patch clamp technique. At the same time, intracellular calcium concentration ([Ca2+]i) was monitored via the fluorescent calcium-indicator dye Fura-2, which was loaded into cells by diffusion from the patch pipette. 2. Stimulation of mast cells with secretagogues, such as compound 48/80 or substance P, caused release of Ca2+ from internal stores. In addition, external agonists also induced influx of external calcium in 26% of the cells investigated. The agonist-stimulated Ca2+ influx was increased during membrane hyperpolarization and was associated with small whole cell currents. 3. Likewise, internal application of inositol 1,4,5-trisphosphate (Ins1,4,5P3:0.5-10 microM) elevated [Ca2+]i due both to release of Ca2+ from internal stores and to influx of external calcium. The Ins1,4,5P3-induced influx was greater at more negative membrane potentials, suggesting that Ins1,4,5P3 opened a pathway through which calcium could enter at a rate governed by its electrochemical driving force. 4. Inositol 1,3,4,5-tetrakisphosphate (Ins1,3,4,5P4) did not induce Ca2+ influx by itself nor did it facilitate or enhance Ins1,4,5P3-induced Ca2+ entry. Calcium influx was also induced by inositol 2,4,5-trisphosphate. Since this inositol phosphate is a poor substrate for Ins1,4,5P3 3-kinase it seems unlikely that Ins1,3,4,5P4 plays a role in the regulation of the Ca2(+)-influx pathway in mast cells. 5. The Ins1,4,5P3-induced Ca2+ influx was associated with whole-cell currents of 1-2 pA or less, with no channel activity detectable in whole-cell recordings. The small size of the whole cell current suggests either that the Ins1,4,5P3-dependent influx occurs via small-conductance channels that are highly calcium specific or that the influx is not via ion channels. 6. Agonist stimulation also activated large-conductance (ca 50 pS) cation channels, through which divalent cations could permeate; thus, these channels represent a second pathway for Ca2+ influx. The slow speed of activation of the channels by agonists, their activation by internal guanosine 5' O-(3-thiotriphosphate) (GTP-gamma-S), and the inhibition of agonist activation by internal guanosine 5'-O-(2-thiodiphosphate) (GDP-beta-S) all suggest that the 50 pS channels are regulated by a second messenger and/or a GTP-binding protein. The activity of the 50 pS channel in mast cells is not sensitive to either Ins1,4,5P3 or Ins1,3,4,5P4. Activity of the channel was inhibited by elevated [Ca2+]i.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2559967 TI - Characterization of inhibition mediated by adenosine in the hippocampus of the rat in vitro. AB - 1. Intracellular recordings with single-electrode voltage clamp were employed to study the mechanism of adenosine-elicited inhibition of CA1 neurones of the rat in vitro. 2. Adenosine elicits a steady-state outward current in association with an increase in conductance. The driving force varied with external potassium concentration as predicted by the Nernst equation for a change primarily in potassium permeability. 3. Adenosine current was blocked by high concentrations of 4-aminopyridine or barium. In the majority of neurones this current was voltage insensitive. In the remainder, the current was inwardly rectifying. The rectification was blocked by tetraethylammonium. 4. When the adenosine-elicited potassium current was blocked, slow inward currents, normally carried by calcium, were unaffected by adenosine. We conclude that this adenosine inhibition is mediated by an increase in a voltage- and calcium-insensitive potassium conductance in CA1 neurones. PMID- 2559969 TI - Chloride conductance activated by external agonists and internal messengers in rat peritoneal mast cells. AB - 1. Stimulation of mast cells by externally applied secretagogues activated a slowly developing membrane current. With high external and low internal chloride (Cl-) concentrations, the current reversed at about -40 mV, but when external Cl- was made equal to internal Cl-, the reversal potential shifted to about 0 mV, demonstrating that the current carrier was Cl-. 2. In addition to external agonists, internally applied cyclic AMP and high concentrations of intracellular calcium [Ca2+]i could also activate the Cl- current. However, elevated [Ca2+]i produced only slow and incomplete activation. This suggests that the Cl- current is not directly Ca2+ activated. Also, activation of Cl- current by external agonists and by cyclic AMP was unimpaired when [Ca2+]i was clamped to low levels with internal ethylene glycol bis-N,N,N',N'-tetraacetic acid (EGTA), indicating that elevated [Ca2+]i is not necessary for activation of the Cl- current. Although activation by cyclic AMP was faster than that produced by elevated [Ca2+]i, it still required tens of seconds; thus the effect of cyclic AMP was also likely to be indirect. 3. Internal guanosine 5'-O-(3-thiotriphosphate) (GTP gamma-S) could also activate the Cl- current, suggesting the involvement of a G protein in the control of the current. 4. The variance associated with the Cl- current was small, and noise analysis gave a lower limit of about 1-2 pS for the single-channel conductance. The Cl- current was reduced by 4,4'-diisothiocyano 2,2'-stilbenedisulphonate (DIDS), and during DIDS blockade, the variance of the current increased. This suggests that DIDS enters and blocks the open channel. 5. Activation of the Cl- current would make the membrane potential negative following stimulation of a mast cell, thus providing a driving force for entry of external calcium via the stimulation-induced influx pathways described in the preceding paper (Matthews, Neher & Penner, 1989). PMID- 2559970 TI - Adrenal responses to splanchnic nerve stimulation in conscious calves given naloxone. AB - 1. The effects of stimulating the peripheral end of the right splanchnic nerve in the presence of naloxone (2 mg kg-1) have been investigated in conscious 3 to 6 week-old calves. 2. Mean aortic blood pressure rose to significantly higher levels during splanchnic stimulation in bursts at 40 Hz for 1 s at 10 s intervals than it did during stimulation at the corresponding continuous frequency (4 Hz). Furthermore, naloxone significantly reduced the fall in mean vascular resistance in response to both patterns of stimulation. 3. The output of catecholamines from the adrenal gland, together with the proportion of noradrenaline released, was significantly enhanced by stimulating the splanchnic nerves in bursts in animals pre-treated with naloxone and the proportion of noradrenaline released also increased. In both cases the output of adrenaline and noradrenaline was within the same range as that reported previously in normal control animals. 4. Naloxone significantly increased the amounts of enkephalin-like immunoreactivity and corticotrophin-releasing factor (CRF)-like immunoreactivity released from the adrenal gland in response to splanchnic nerve stimulation and raised the proportion of total to free met5-enkephalin that was secreted. 5. Naloxone also inhibited the rise in plasma adrenocorticotrophic hormone (ACTH) concentration during continuous stimulation at 4 Hz, but not during stimulation at 40 Hz in bursts. Under these latter conditions the output of cortisol apparently directly from the adrenal gland was inhibited. The finding that splanchnic nerve stimulation can potentiate the output of cortisol in response to ACTH was confirmed. 6. These results provide evidence that release of enkephalins and of CRF from the adrenal is inhibited by activating opioid receptors within the gland itself. PMID- 2559972 TI - Transient and persistent sodium currents in normal and denervated mammalian skeletal muscle. AB - 1. Transient and persistent tetrodotoxin-sensitive sodium currents were recorded in response to depolarizing voltage pulses in voltage clamped segments of rat extensor digitorum longus muscle fibres at 20-25 degrees C in a triple Vaseline gap. 2. Appreciable persistent sodium current but little or no transient current was seen in response to depolarizations of up to 15 mV from a holding potential of -100 mV. 3. The maximum amplitude of both transient and persistent sodium currents occurred with depolarizations to -40 mV: the average peak amplitude of the transient current in fibres with a holding potential of -90 mV was -0.22 +/- 0.03 mA/microF (mean +/- 1 S.E.M., seven fibres) and the average amplitude of the persistent current was -0.94 +/- 0.10 microA/microF (mean +/- 1 S.E.M., twelve fibres). With a holding potential of -100 mV, the average amplitudes of the transient and persistent currents were -0.46 +/- 0.10 mA/microF (four fibres) and -1.4 +/- 0.22 microA/microF (five fibres), respectively. 4. The average maximum persistent sodium conductance in seven fibres held at -90 mV was 0.13 +/- 0.0078 microS and the potential for half-maximum conductance was -53 +/- 0.74 mV (mean +/- 1 S.E.M.). 5. When the transient sodium current was completely inactivated with 100 ms conditioning depolarizations to potentials more positive than -50 to 60 mV, there was little inactivation of the persistent current. 6. In six denervated fibres, the average amplitudes of the transient and persistent sodium currents generated by pulses to -40 mV from a holding potential of -90 mV were 0.11 +/- 0.01 mA/microF and -0.88 +/- 0.12 microA/microF, respectively (mean +/- 1 S.E.M.). It was concluded that there was a decrease in transient current but not persistent current amplitude following denervation and that the persistent current in denervated fibres with an increased input resistance could give rise to the spontaneous action potentials responsible for fibrillation. PMID- 2559971 TI - Calcium channels in solitary retinal ganglion cells from post-natal rat. AB - 1. Calcium currents from identified, post-natal retinal ganglion cell neurones from rat were studied with whole-cell and single-channel patch-clamp techniques. Na+ and K+ currents were suppressed with pharmacological agents, allowing isolation of current carried by either 10 mM-Ca2+ or Ba2- during whole-cell recordings. For cell-attached patch recordings, the recording pipette contained 96-110 mM-BaCl2 while the bath solution consisted of isotonic potassium aspartate in order to zero the neuronal membrane potential. 2. A transient component, present in approximately one-third of the whole-cell recordings resembles closely the T-type calcium current observed previously in other tissues. This component activates at low voltages (-40 to -50 mV from holding potentials negative to -80 mV), inactivates with a time constant of 10-30 ms at 35 degrees C, and is carried equally well by Ba2+ or Ca2+. In single-channel recordings small (8 pS) channels are observed whose aggregate microscopic kinetics correspond well to the macroscopic current obtained during whole-cell measurements. 3. During whole-cell recordings, a more prolonged component activates in all retinal ganglion cells at -40 to -20 mV from a holding potential of -90 mV. This component is substantially larger when equimolar Ba2+ replaces Ca2+ as the charge carrier, and is sensitive to the dihydropyridine agonist Bay K8644 (5 microM) and antagonists nifedipine (1 10 microM) and nimodipine (1-10 microM). Thus, the dihydropyridine pharmacology of this prolonged component resembles that of the L-type calcium current found in dorsal root ganglion neurones and in heart cells. Also reminiscent of the L current, the prolonged component in this preparation is less inactivated at depolarized holding potentials (-60 to -40 mV) than the transient component. In cell-attached recordings, large (20 pS) channels are observed with activation properties similar to those of the prolonged portion of the whole-cell current. 4. omega-Conotoxin fraction GVIA (omega-CgTX VIA), a peptide from the venom of the snail Conus geographus, produces a readily reversible blockade of all components of the calcium current in these central mammalian neurones. This finding is in contrast to that of other preparations in which this toxin is responsible for an ephemeral block of T-current but a long-lasting block of other components of calcium current. 5. In summary, at least two components of calcium current with discrete underlying unitary events are present in post-natal retinal ganglion cells from rat. One component closely resembles the T or transient current observed in other cell types.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2559973 TI - GABA and glycine channels in isolated ganglion cells from the goldfish retina. AB - 1. Adult goldfish retinas were enzymatically dissociated and ganglion cells were maintained in culture for periods of 1-5 days. Ganglion cells could be identified by their morphology, and this identification was confirmed by retrograde transport of the fluorescent dye Fast Blue injected into the optic nerve stub. 2. All the ganglion cells tested responded to 30 microM-GABA or 100 microM-glycine between 2 and 30 h after enzymatic dissociation of the retina. 3. Whole-cell responses to 30 microM-GABA or glycine declined over a period of seconds during sustained applications of the agonists, probably as a result of desensitization. There was an irreversible decline in the peak whole-cell response to repeated applications of 30 microM-GABA unless the pipette-filling solution contained 2 mM ATP, 4 mM-Mg2+, 10 mM-EGTA and no added Ca2+. Both GABA and glycine responses also showed an irreversible decline in outside-out patches but, in this case, Mg2+, ATP, and very low Ca2+ failed to stabilize the response. 4. Whole-cell currents activated by both GABA and glycine were demonstrated to be chloride selective by investigating the dependence of reversal potential (Vr) on internal chloride concentration ([Cl-]i). For GABA responses, the dependence of Vr on [Cl ]i could not be distinguished from that predicted by the Nernst relation. For glycine, deviations from Nernstian dependence were observed, but the permeability to Cl- was at least 20 times greater than to isethionate, SO4(2-), or monovalent cations (Na+ and Cs+). 5. Bicuculline methochloride (10 microM) selectively blocked responses to 3-30 microM-GABA without affecting responses to 30 microM glycine. Bicuculline itself was not as selective. At agonist concentrations of 30 microM, 3 microM-bicuculline partially blocked the response to GABA but not that to glycine, but bicuculline at 10 microM blocked responses to both GABA and glycine. Strychnine (0.3-1 microM) blocked responses to 30 microM-glycine but also competitively antagonized GABA responses. Picrotoxinin (10 microM) blocked responses to 3 microM-GABA in some cells but also partially antagonized responses to 30 microM-glycine. 6. GABA channels had at least two conductance states at 10 12 degrees C in nearly symmetrical (141 mM in, 142 mM out) chloride. The slope conductance of the most frequently observed (main) state was 16 +/- 2 pS. The reversal potential for the main state was not significantly different from the chloride equilibrium potential (0 mV).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2559974 TI - Role of central opiate receptor subtypes in the circulatory responses of awake rabbits to graded caval occlusions. AB - 1. In unanaesthetized rabbits, haemorrhage was simulated by inflating a cuff placed round the inferior vena cava so that cardiac output fell at a constant rate of approximately 8% of its resting value per minute. The circulatory responses were measured after injections into the fourth ventricle of saline vehicle, selective opioid antagonists, selective opioid agonists, and agonist antagonist mixtures. Three sets of experiments were done to determine if a specific subtype of opiate receptor within the central nervous system is responsible for the circulatory decompensation that occurs during simulated haemorrhage. 2. In six rabbits the effects of ascending doses of the antagonists naloxone (mu-selective), Mr 2266 (kappa- and mu-selective), ICI 174864 (delta selective) and nor-binaltorphimine (kappa-selective) were tested. In three rabbits the effects of the antagonist naloxone, the agonists HTyr-D-Ala-Gly-MePhe NH(CH2)2OH (DAGO, mu-selective), U 50488H (kappa-selective), and [D-Pen2,D-Pen5] enkephalin (DPDPE, delta-selective), and combinations of these agonists with naloxone were tested. In four rabbits the dose-related effects of DAGO on respiratory, as well as circulatory, functions were examined. 3. After injecting saline vehicle, the circulatory response to simulated haemorrhage had two phases. During the first phase, systemic vascular conductance fell, heart rate rose, and mean arterial pressure fell by only approximately 10 mmHg. A second, decompensatory, phase began when cardiac output had fallen to approximately 50% of its resting level. At this point, there was an abrupt rise in systemic vascular conductance and a fall in mean arterial pressure to less than or equal to 40 mmHg. 4. The lower range of doses of naloxone (3-30 nmol), Mr 2266 (10-100 nmol), ICI 174864 (10-30 nmol), and all doses of nor-binaltorphimine (1-100 nmol), were without effect on the circulatory response to stimulated haemorrhage. Higher doses of naloxone (30-100 nmol), Mr 2266 (100-300 nmol) and ICI 174864 (30 100 nmol) abolished the decompensatory phase. The relative order of antagonist potency was ICI 174864 greater than or equal to naloxone greater than Mr 2266 greater than or equal to nor-binaltorphimine. 5. In the second set of experiments, the critical dose of naloxone necessary to prevent circulatory decompensation during simulated haemorrhage was 30-150 nmol. The delta-agonist DPDPE (50 nmol) did not affect the haemodynamic response to simulated haemorrhage, but it did block the effect of naloxone on the response.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2559975 TI - Decoupling of horizontal cells in carp and turtle retinae by intracellular injection of cyclic AMP. AB - 1. Horizontal cells are electrically coupled through gap junctions. This is a disadvantage in elucidating the membrane properties of the cells. In order to block gap junctions, adenosine 3',5'-cyclic monophosphate (cyclic AMP) or its analogues, dibutyryl cyclic AMP and 8-bromo cyclic AMP, were ionophoretically injected into horizontal cells of the carp or turtle retina. 2. Before injection of the chemicals the input resistance of the cell was so low as to be unmeasurable, because the applied current leaked through gap junctions. After injection, however, the input resistance was significantly increased. 3. After the injection dye-coupling between horizontal cells was not observed when examined by intracellular injection of Lucifer Yellow dye, supporting the idea that high concentrations of intracellular cyclic AMP block gap junctions. 4. In this situation responses to light delivered to the receptive field centre were increased in amplitude, while responses to light delivered to the receptive field surround were greatly diminished. 5. After injection horizontal cells were readily polarized by conventional intracellular current injection. The hyperpolarizing light responses in carp and turtle luminosity-type cells (H1 cells) could be reversed by depolarizing the horizontal cells, and the reversal potentials were estimated to be about 0 mV. In addition, the resistance increase which accompanied the hyperpolarizing light responses could be detected. 6. In turtle biphasic chromaticity-type horizontal cells (H2 cells), hyperpolarizing light responses to shorter wavelengths and depolarizing ones to longer wavelengths could be reversed by depolarizing the horizontal cells. Both responses have almost the same reversal potential at about 0 mV. The membrane resistance changes associated with light responses were also detected; the resistance increased during the hyperpolarizing response, while it decreased during the depolarizing response. These observations suggest that the ionic mechanisms of both responses are probably the same, irrespective of their polarities. PMID- 2559976 TI - Kinetic properties of the cardiac T-type calcium channel in the guinea-pig. AB - 1. The kinetic properties of T-type Ca2+ channels were examined in single ventricular cells from guinea-pig hearts using the cell-attached configuration of the patch-clamp technique. 2. T-type Ca2+ channel activity has been observed in 44 out of 139 patches. The density of these channels was estimated at 0.1-0.3 micron-2. The T-type Ca2+ channel responds to a depolarizing voltage step either with a burst of openings which appears with a distinct delay or with no openings at all. The mean number of bursts per record for the records showing channel activity is 1.1. The probability of observing a blank sweep is high and amounts to 0.65 +/- 0.02 (n = 26). 3. With 110 mM-Ca2+ in the pipette solution, the slope conductance calculated from the current-voltage relationship of the single channel current in the range between -50 and +10 mV is 6.8 pS. 4. Openings to a subconductance level of about 50% of the main level could be resolved. All possible transitions between the subconductance and the main level were observed, indicating that the cardiac T-type Ca2+ channel possesses a substate. 5. The macroscopic steady-state activation and inactivation, as determined from ensemble averaged currents, could be described by Boltzmann functions. Half-maximal activation and inactivation occur at -14 and -60.7 mV, the slope parameters of these curves are 10.8 and 5.6 mV respectively. The maximum (peak) open probability is 0.15. 6. The ensemble-averaged current decays monoexponentially. The time constant is strongly voltage dependent and decreases at less negative potentials. 7. The open times are monoexponentially distributed. The mean open time of the channel does not depend on either the holding or the test potential, and has a mean value of 1.4 ms. The distribution of the closed times is biexponential. The fast mean closed time is also voltage independent with a mean value of 0.48 ms. The slow mean closed time increases with voltage from 1.9 ms at -40 mV to 8.8 ms at 0 mV. The mean burst duration also increases with voltage from a value of 4.9 ms at -40 mV to 13.9 ms at -10 mV. 8. The convolution of the first-latency distribution with that of the burst duration closely fits the open probability calculated from the ensemble-averaged current. The mean first latency is also closely correlated with the macroscopic time constant of inactivation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2559977 TI - ATP-activated channels gate calcium entry in single smooth muscle cells dissociated from rabbit ear artery. AB - 1. A combination of the techniques of microspectrofluorimetry and whole-cell patch clamp was used to investigate changes in cytoplasmic Ca2+ concentration (Cai2+) in single arterial smooth muscle cells on external application of ATP. 2. ATP applied to cells held under voltage clamp at --60 mV evoked an inward current and an associated rise in Cai2+. In the absence of extracellular Ca2+. ATP activated inward currents were observed but there was no rise in Cai2+. 3. Pre treatment of cells with noradrenaline or caffeine did not prevent the rise in Cai2+ on subsequent application of ATP. 4. The ATP-activated rise in Cai2+ was voltage dependent as outward currents evoked by ATP at positive membrane potentials were not associated with a change in Cai2+. 5. At --60 mV, the rise in Cai2+ due to ATP application was dependent on the magnitude of the ATP current response, such that Cai2+ increased by about 0.5 nM/pC charge transferred through ATP-gated channels. 6. The results suggest that ATP-gated channels in these cells admit sufficient Ca2+ in a physiological Ca2+ gradient to significantly elevate Cai2+. About 10% of the ATP-gated current may be carried by Ca2+ ions. Thus the ATP-activated channels have a dual excitatory function: depolarization due to Na+ entry promotes action potential discharge and voltage-gated Ca2+ entry, and also direct entry of Ca2+ through the ATP-activated channels. PMID- 2559979 TI - Fine structure of EBV-infected keratinocytes in oral hairy leukoplakia. AB - We evaluated biopsy specimens of 42 cases of clinically suspected oral hairy leukoplakia for the pattern and frequency of ultrastructural alterations specific to epithelial cells infected with Epstein-Barr virus. Some structures could clearly be identified as Epstein-Barr virus at different stages of assembly, but other intranuclear and cytoplasmic alterations were not conclusively identifiable as any known structure. Keratinocytes producing Epstein-Barr virus contained intranuclear particles of different size and shape; some of them were arranged in a monodispersed pattern and others formed arrays. In contrast, both lesional keratinocytes not producing virus and keratinocytes in uninvolved mucosa contained intranuclear particles reminiscent of perichromatin granules. The nuclei of productive cells also contained marginated chromatin, tubular structures, and, occasionally, crystalline and fibrillar formations as well as enveloped virus. Formations of electron-dense bilayers were seen on both sides of the nuclear membrane. In the cytoplasm of productive cells we observed aggregates of parallel tubules and enveloped electron-dense bodies. Although many of these observations are of diagnostic and pathobiological significance, the morphogenesis, composition, and function of alterations with uncertain morphological identification remain unclear. PMID- 2559981 TI - Cystosarcoma phyllodes. AB - Ten cases of cystosarcoma phyllodes are reported along with review of literature. The pathological features are outlined and principles of management are discussed. PMID- 2559980 TI - Iron deficiency reduces cytochrome concentrations of mitochondria isolated from hamster cheek pouch epithelium. AB - The effects of iron deficiency on the aerobic pathway of energy metabolism were studied using mitochondria isolated from epithelial cells from the hamster cheek pouch. A statistically significant reduction in the concentrations of cytochromes aa3, b and c (P less than 0.05), a reduction (P = 0.064) in cytochrome cl and altered cytochrome ratios were found in the mitochondria of iron deficient compared to normal animals. State 4 respiration was demonstrated in the mitochondria of both normal and iron deficient animals but state 3 respiration could not be demonstrated; this suggests uncoupling of oxidative phosphorylation which may be an artefact associated with the separation of epithelium from its connective tissue. Nevertheless we conclude that the reduction in cytochrome concentration is a real effect of iron deficiency which may explain, at least in part, the reduction of both energy production and cell proliferation seen in oral epithelia under these conditions. PMID- 2559978 TI - cis-Fatty acids, which activate protein kinase C, attenuate Na+ and Ca2+ currents in mouse neuroblastoma cells. AB - 1. Activation of protein kinase C (PKC) by phorbol esters or diacylglycerols has been shown to modulate a number of ionic currents carried by Ca2+, K+ and Cl-. Recently, it has been demonstrated that PKC may be activated by cis-fatty acids in the absence of either phospholipid or Ca2+. We wished to determine if this new class of PKC-activating compound would also modulate ionic currents. To this end we applied the whole-cell voltage-clamp technique to N1E-115 neuroblastoma cells. 2. Analysis of families of currents evoked under voltage clamp by depolarizing steps from a holding potential of -85 mV during external application of 5 microM oleate (a cis-fatty acid) showed a 36% reduction of the peak inward current with no shift in either the peak or the reversal potential of the current-voltage relation and no alteration of outward current. 3. External application of the cis fatty acids oleate, linoleate and linolenate reversibly attenuated voltage dependent Na+ current with approximate half-maximal dose values of 2, 3, and 10 microM respectively. Oleate was approximately 2 times more potent when applied internally (ED50 = 1 microM). Externally applied elaidate (a trans-isomer of oleate) and stearate (a saturated fatty acid) which do not activate PKC, had no effect. Since cis-fatty acids are known to fluidize membranes, as well as to activate PKC, we sought to dissociate these functions by applying compounds that fluidize membranes but do not activate PKC: methyloleate and lysophosphatidylcholine. Neither compound affected Na+ current when applied externally at concentrations of 1-50 microM. 4. In contrast to cis-fatty acids, three classical PKC activators, phorbol-12.13-dibutyrate (PDB), phorbol-12.13 diacetate (PDA), and 1.2-oleoylacetylglycerol (OAG) were found to have no effect on the voltage-dependent Na+ current when applied externally at 10 nM-1 microM (phorbol esters) or 1-150 microM (OAG) for incubation periods up to 1 h. 5. External application of the PKC inhibitors polymyxin B, H-7, sphingosine and staurosporine blocked the attenuation of the Na+ current by cis-fatty acid in a dose-dependent manner, with maximal inhibition occurring at doses of 50, 10, 200 and 0.1 microM, respectively. The cyclic nucleotide-dependent protein kinase inhibitor H-8 was much less effective in blocking the cis-fatty acid effect. Polymyxin B and staurosporine were more potent when applied internally. 6. Chronic (24 h) exposure to 1 microM phorbol-12-myristate-13-acetate (TPA) was employed to down-regulate PKC.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2559983 TI - [Nonfunctioning nesidioblastoma of the pancreas in children. Apropos of a case]. AB - A new case of nonfunctional nesidioblastoma in a 13-year-old girl is reported. The diagnosis was made with imaging, including radiographs of the abdomen, ultrasound and computed tomography. Complete resection of the pancreatic tumor was possible. The patient has no symptoms 8 months later. PMID- 2559982 TI - [A rare pulmonary tumor: pneumoblastoma. Apropos of a case and review of the literature]. AB - A case of pneumoblastoma concerning a 33 year old man, incidentally discovered on a systematic chest X Ray, is reported. The radiologic findings include a voluminous and peripherical mass with necrotic zones. The pathologic findings in optic microscopy and the special technics consist in the three cellular contingents which form the pneumoblastoma: clusters of indifferentiated cells, epithelial cells and spindle cells. All the features show clearly a blastomatic character very similar to a foetal lung which allows to differentiate the pulmonary blastoma from carcinosarcoma. The authors, through a review of the literature, demonstrate the rarity (less than 100 cases reported), the malignant evolution which metastasis and the age (strong majority of adults) which characterize this pathology. PMID- 2559984 TI - [Abrikossoff's tumor with bronchial localization. Contribution of x-ray computed tomography in a new case]. AB - The authors present a case of Abrikossoff's tumor located in the segmental bronchus and detected on a systematic chest radiography, diagnosed during an endoscopy is reported. The CT study specified the tumoral extension. Bronchial distal affections is very rare. A confrontation with the anatomopathological data allowed a better understanding scanographic aspect. PMID- 2559985 TI - Direct access to hospital investigative facilities: threats and opportunities. PMID- 2559986 TI - Is paediatrics safe in general practitioners' hands? A study in the north of England. AB - More than half the general practitioners in the north of England, and all the health visitors, were sent questionnaires about the primary health care of children. Eighty per cent of general practitioners responded. Their qualifications and experience in paediatrics were poor, although this was less true for younger practitioners. The doctors differed widely in their management of hypothetical clinical problems, possibly owing to this lack of training. Health visitors were thought to have an important role in developmental screening within the primary health care team, but the teamwork itself seemed weak. Although 83% of general practitioners considered developmental screening should be primarily the responsibility of the primary health care team, 31% of practices did not do it and had no plans to start. Rather more than half of the doctors in these non-screening practices would accept a clinical medical officer to screen their patients on their premises. Suggestions are made for ways of encouraging general practitioners to carry out developmental screening. PMID- 2559987 TI - Attitudes and practices of the primary health care team towards assessing the very elderly. AB - This paper investigates the attitudes and practices of members of the primary care team towards assessing the very elderly in the community. Using self completion questionnaires 47 general practitioners, 24 health visitors and 22 community nurses in the Bath health district were asked if they felt over 75 year olds should be assessed, and if so who should do it, and what the assessment should include. A majority of all three groups of professionals considered it important to assess the over 75 year olds and that this should be done at home. They felt that such an assessment should be functionally oriented, that is, should concentrate on the activities of daily living rather than medical problems. None of the groups felt assessment was their sole responsibility but should be undertaken by a combination of members of the primary health care team with the health visitor playing a prominent role. Although 89% of all the respondents felt that health visitors should be involved in assessing the elderly, half of the health visitors felt their work should be mainly concerned with 0-5 year olds. In the absence of any general policy few assessment schemes exist in the district. An overall policy is required and this should be reflected in the training of each of the professional groups. PMID- 2559988 TI - Receptionists, appointment systems and continuity of care. AB - The influence of receptionists on continuity of care in four group practices was examined. Twenty two receptionists were observed making 543 appointments and afterwards were interviewed about their personal priority for continuity of care. One of the practices ran a personal list system. It was found that the personal list practice attracted more requests for a specific doctor and time which were then more difficult to satisfy. Most receptionists thought it was important that patients should see the same doctor but their influence seemed to be small compared with that of the doctor as expressed in practice policies. Marked variation in demand for individual doctors was seen in two of the practices without a personal list system. PMID- 2559989 TI - Survey of general practitioners' advice for travellers to Turkey. AB - Fifty general practitioners replied to a survey of the advice that they would offer to a tourist planning a package holiday in western Turkey. The range of prophylactic immunizations and other medication recommended by the respondents was wide, suggesting that some tourists travel without adequate protection, while some receive unnecessary injections. Most of the doctors would offer little other health advice to the traveller. General practitioners receive conflicting guidance on prophylactics for travellers, and it is suggested that the disagreements should be resolved. Wider availability of written advice for the traveller would also be valuable. PMID- 2559990 TI - Misplaced loss of confidence in measles vaccination: an investigation in a primary school. AB - Doubts were expressed about the effectiveness of measles vaccination during a measles outbreak in a Bristol primary school. Investigation of this outbreak showed that the vaccine uptake rate was 89%, vaccine effectiveness was 84% and that 57% of children developing measles had a history of previous measles vaccination. This apparent paradox arises because, for any vaccine that is not 100% effective, as the vaccine uptake rate rises so the percentage of cases previously vaccinated also rises. An understanding of this relationship may avoid unjustified loss of confidence in vaccine effectiveness as vaccine uptake increases. PMID- 2559991 TI - Patients' choice of general practitioner: influence of patients' fluency in English and the ethnicity and sex of the doctor. AB - Asian patients' use of general practitioner services and, in particular, their interaction with doctors is not well researched. However, difficulty in communication and in the case of women, reluctance to be examined by a male doctor has been reported. This study, based on interviews with 241 Caucasian, Pakistani and Indian patients attending a general practice in Bradford, examined the relationship between choice of general practitioner and the patient's fluency in English and the general practitioner's ethnicity and sex. Both Pakistani and Indian patients, particularly women, had poor fluency in English and the use of interpreters was confined to women (11% of Pakistani women and 4% of Indian women). The linguistic and broad cultural concordance between the patient and the general practitioner was more important in the choice of doctor than the sex of the general practitioner. It was also found that while 62% of Pakistani women objected to being examined by a male doctor, this was true for only 21% of Indian women. PMID- 2559993 TI - Sudden death after spontaneous ventricular fibrillation: case reports. AB - Patients resuscitated from ventricular fibrillation in the absence of myocardial infarction remain at high risk of sudden death, even if underlying risk factors are treated. Two such cases are reported here. PMID- 2559992 TI - Perceived versus actual consultation patterns in an inner city practice. AB - Consultation patterns in a practice with no appointment system, situated in a socially deprived area, were examined to confirm or refute one doctor's perception that the consultation rate was much higher than average. Seventy five per cent of a sample of patients (n = 394) had consulted in the previous year. The mean annual consultation rate was 3.8 (range 0 to 29) but the median was 3.0 and the mode 0. Among 222 patients consulting over one month the mean annual consultation rate was 10.0 (range 0 to 47) with a median of 9.0 and a mode of 6. The duration of consultation (n = 506) varied from one to 25 minutes (with mean, median and modal values of 5.3, 4.0 and 4 minutes respectively) and 53% of patients received between one and four minutes. The overall consultation rate was not high, particularly in view of the socioeconomic deprivation of the practice population, and the doctor's perception of excessive consultation was explained by the high consultation rate among attenders. The consultation pattern, particularly of males, was not conducive to a preventive approach. The mean was a poor descriptor of the average consultation rate or duration. PMID- 2559995 TI - The Christian approach to whole-person medicine. AB - A working party with representatives from the Royal College of General Practitioners and the Churches' Council for Health and Healing has produced a report on the Christian approach to whole-person medicine. This emphasizes the importance of including the spiritual dimension of man in health care problems and urges closer cooperation between health care professionals and Christian ministers. Some of the difficulties inherent in this approach are highlighted in this paper and some proposals for further progress outlined. PMID- 2559994 TI - Primary and community health services in Newcastle upon Tyne--a joint statement of intent. AB - In the absence of a single primary health care authority (except in Scotland) district health authorities and family practitioner committees must cooperate in planning health services for the community. Equally, in the field, the potential for teamwork between salaried district health authority nursing staff and the independent general practitioners remains largely unrealized. Yet the government has restated its commitment to the development of primary health care teams as the best means of delivering health care in the community. In Newcastle upon Tyne the local medical committee and the community health services management team have set out their shared aspirations for future development in a joint 'Statement of intent'. This statement, since endorsed by the district health authority and family practitioner committee, includes a number of key principles as a basis for future joint working. These principles emanate from an understanding of the complementary nature of general practice and district health authority community services, and firmly support the primary health care team approach. This statement of intent could serve as a useful model for collaboration and planning of services elsewhere in the country. PMID- 2559996 TI - Working against patients? PMID- 2559997 TI - Management of chronic (post-viral) fatigue syndrome. PMID- 2559998 TI - Treatment of warts with liquid nitrogen. PMID- 2559999 TI - Breast self examination: should we discourage it? PMID- 2560000 TI - Randomized controlled trial of small group education on the outcome of chronic asthma in general practice. AB - The effectiveness of small group education of general practitioners in the management of asthma was evaluated by randomized controlled trial. The outcome measure was the asthma morbidity of the general practitioners' own patients. Following random selection from the list of one family practitioner committee in suburban London, the 27 participating general practitioners were allocated randomly to one of two educational groups or to a control group. The educational intervention comprised eight meetings at which the management of chronic asthma was discussed and attempts made to devise agreed strategies for care. The two educational groups devised different strategies. Asthma morbidity was assessed by postal questionnaires to patients before the intervention and on five further occasions at six-monthly intervals over two and a half years. Of 454 patients who entered the study 338 completed the sixth and final assessment. The degree of morbidity experienced by the patients and their reported use of asthma specific drugs was considerable and was notably constant over the period of study. There was no difference in morbidity between the three groups at the outset and no effect of the intervention could be demonstrated. In this educational intervention the participating general practitioners were not informed about the morbidity and drug use reported by their patients. This information may be crucial if small groups are to be used to design and implement effective strategies for care. It would appear that small group education of general practitioners in the form reported here is not effective in reducing morbidity from chronic asthma. PMID- 2560001 TI - General practitioners and their learning styles. AB - Continuing medical education sessions are often poorly attended by general practitioners. One reason may be that these traditionally consist of lectures by hospital consultants with a strong theoretical bias which may have little relevance to the learning needs of general practitioners. To compare the learning styles of teachers and learners in general practice, learning style questionnaires were administered to 50 hospital clinical tutors, 78 general practitioner trainers, 63 trainees and 47 non-trainer principals. The questionnaire covered four different learning preferences: activist, reflector, theorist and pragmatist. The findings showed that the learning styles of hospital tutors and general practitioner trainers were statistically significantly different to those of non-trainer principals and trainees. The tutors and trainers scored much higher on theorist styles and to a lesser extent on reflector and pragmatist styles. There were no significant differences on activist scores. Since teachers tend to teach in their preferred learning style, which may not match the style of the recipients, these findings have implications for continuing medical education in general practice. These implications are discussed. PMID- 2560002 TI - Continuing educational requirements for general practitioners in Grampian. AB - A postal questionnaire about educational requirements achieved an 81% response from all general practitioners in Grampian. A majority of respondents felt that it was important to keep up-to-date and they were setting aside a number of hours per week to do this. Evening meetings, intensive two to three day courses and week long refresher courses were still popular, as was the idea of small groups resourced by a consultant. Newer forms of continuing education, for example, audio and videotape and distance learning were not popular. Journals still remained an important way of keeping up-to-date. These results help in planning the rational use of postgraduate resources for the future. PMID- 2560003 TI - Can health screening damage your health? AB - This study set out to determine whether screening can be psychologically harmful to healthy adults. A prospective controlled study was carried out on 215 healthy adults attending a by-invitation coronary heart disease screening clinic in general practice. The general health questionnaire was used as an indicator of recent psychological distress. Patients attending the screening clinic had significantly lower subjective psychological distress than an unscreened group of 225 age-matched controls, indicating that we may well be screening an already psychologically healthy sub-group. The main finding was that patients' own assessment of their psychological distress was significantly increased three months after screening compared with that of controls, who showed a non significant decrease. It is concluded that there is a real risk of causing distress by screening healthy adults and that this possibility has largely been ignored by previous studies. Possible explanations and implications of these findings are discussed, particularly in the light of increased pressure from many quarters for more screening services to be set up in general practice. PMID- 2560004 TI - Opportunistic health promotion: quantity or quality? AB - A cohort of 130 working class mothers has been studied in depth over five years to quantify the extent of recording and counselling of lifestyle problems by general practitioners and their staff. Clinical records and mothers' personal accounts at two home interviews five years apart provide the data for this work. Fifty-nine per cent of women had one or more aspects of lifestyle recorded in their records, the commonest being smoking habits. Despite this evidence for good coverage of smokers in the population, alcohol and exercise problems were under recorded. Clinical records only included details of advice given and follow-up plans for lifestyle problems in 40% of patients' records yet the women themselves remembered advice being given in 48% of cases. An analysis of the womens' accounts in conjunction with the clinical records revealed that over three quarters of those receiving advice remembered it several years later. The primary care team was most likely to target advice and plans on women who were heavy smokers and very obese. This study shows that clinical records underestimate the amount of lifestyle counselling which is conducted in general practice and that a surprising number of working class women remember and act on the advice from their doctors. The implications for clinical recording of lifestyle factors are discussed. PMID- 2560005 TI - What do health visitors contribute to the care of children? A study in the north of England. AB - All the health visitors in the north of England, and more than half the general practitioners, were sent questionnaires about the primary health care of children. More than 90% of the health visitors responded. Most of them took part in developmental screening and considered it primarily their responsibility; some conducted developmental or well baby clinics with no other professionals present. Clinics run by health authorities often occupied several hours per week, and were more frequently attended by health visitors than clinics run by general practitioners. Almost all the health visitors' remaining time was spent in attached practices, despite the fact that more than half said they had neither office nor clinic space of their own on practice premises. A high proportion of time was spent on clerical work; more help with this could free the health visitor to provide better developmental care for all children. PMID- 2560006 TI - Characteristics and pattern of care of a diabetic population in mid-Wales. AB - The pattern of care and demographic features of diabetes in a largely rural Welsh population were investigated before the introduction of measures to improve diabetic care. All data were obtained from general practice notes. Of the population studied 1.01% were identified as diabetic. There were no demographic differences from recently published English studies. Seventy per cent of the diabetic patients had not seen a consultant within the preceding year. The level of surveillance for biochemical control and complications of diabetes was better in those who had had recent consultant care. However, basic surveillance data was missing in many consultant letters to general practitioners. The prevalence of known serious diabetic eye disease (9%) in the study population was similar to that found in a recent study of a structured care system. Proposed improvements in diabetic care must take into account the large number of patients not attending hospital clinics. Communication between consultant clinics and general practitioners must be improved. PMID- 2560007 TI - The community pharmacist: over qualified dispenser or health professional? AB - This paper contributes to the debate introduced in a previous issue of this journal. The current and future roles of community pharmacists are outlined. It is suggested that although elements of their dispensing role are changing, pharmacists continue to have a vital function in the dispensing process. Proposed developments for the future role of community pharmacists in the treatment of minor illness are also discussed. PMID- 2560008 TI - Neuroleptic malignant syndrome from chlorpromazine: case report. PMID- 2560009 TI - Funds for research. PMID- 2560010 TI - General practice or primary health care? PMID- 2560011 TI - Preventive care card. PMID- 2560012 TI - Management of chronic (post-viral) fatigue syndrome. PMID- 2560013 TI - Role of the community pharmacist. PMID- 2560014 TI - Response to the white paper. PMID- 2560015 TI - Transfer of medical records. PMID- 2560016 TI - Information unbound. PMID- 2560017 TI - Inner cities: primary health care in the market place. PMID- 2560018 TI - Part-time women general practitioners--workload and remuneration. AB - A postal questionnaire survey was conducted comparing the workload and remuneration of part-time women principals in group practices in the Northern and Oxford regions. Part time was defined as receiving less than a full profit share at parity. Of 501 women principals 308 (62%) responded of whom 146 (47%) were part-time. Respondents were asked to record aspects of workload over a four-week period for themselves and their full-time partner who did the most sessions within the practice. The results showed that although two-thirds of the part timers had 50% or less of a full profit share, part-time principals overall did about 76% of the daytime clinical work (surgeries and home visits) done by their full-time partners, excluding specialized clinics. The lower the profit share the wider this discrepancy. Although 33% of the respondents did not out-of-hours work, the remainder did more than their profit share would indicate. Twenty per cent of the 116 principals with 40% or more of a full profit share and 57% of the 30 principals with less than 40% of a full profit share felt that their share was unfair. Lack of involvement in practice business and feeling that opinions did not carry equal weight were associated with feelings of unfairness. PMID- 2560019 TI - Do referral rates vary widely between practices and does supply of services affect demand? A study in Milton Keynes and the Oxford region. AB - Two commonly held beliefs about referral rates were investigated in this study: first that demand for services is determined by supply and secondly that there is wide variation between general practices in their referral rates. All referrals to specialist outpatient clinics were recorded during two 11-week periods by general practitioners in eight practices in the new town of Milton Keynes and in 17 practices elsewhere in the Oxford region. During the first period, only a limited outpatient service was available in the new town; for many specialist services, people had to be referred to hospitals outside the district. Referral rates from Milton Keynes were very similar to those from the rest of the region. By the second period the range of specialist facilities available locally had expanded considerably with the opening of the new district general hospital and during this period there was a statistically significant but rather small increase in referral rates from Milton Keynes. Variation in referral rates between general practices within each geographical group was greater than that between the two groups. Overall, there was about a three-fold variation between general practices in outpatient referral rates which is considerably less than that commonly thought to exist. PMID- 2560020 TI - The effect of minimal interventions by general practitioners on long-term benzodiazepine use. AB - Seventy one long-term users of benzodiazepines were asked by their general practitioners in a letter or short interview to reduce their medication. Twenty two patients were successful in giving up or reducing their consumption to less than 100 tablets per annum. There were no clear predictors of success in terms of patient characteristics, duration of drug use, type of benzodiazepine, reason for drug use or strategy employed to reduce medication. However, patients who were successful at reducing their medication had a significantly lower mean baseline drug consumption than unsuccessful patients. The implications of this study are that a proportion of long-term users who are not in current crisis, especially those with relatively low consumption, can reduce or stop benzodiazepine treatment with minimal difficulty. PMID- 2560021 TI - General practitioners' use of community psychiatric nursing services: a preliminary survey. AB - A questionnaire on general practitioners' use of community psychiatric nursing services was sent to a random sample of 100 general practitioners in two contrasting areas, Croydon and Cambridgeshire. General adult services were widely available though used less often by Cambridgeshire general practitioners than Croyden doctors. Apart from services for the elderly, specialist services were uncommon. Over a third of doctors reported that their adult services were based in a psychiatric hospital. Less than a quarter of general practitioners had access to primary care based nurses. The pattern of responses demonstrates the wide variety of ways in which general practitioners relate to community psychiatric nurses, even where the psychiatric nursing services are long established. There remains a need for more consistent and coherent policies about the ways in which community psychiatric nurses are employed in primary care. PMID- 2560022 TI - Medical screening of 1500 patients in a dental surgery: a prospective study. AB - Communication between medical and dental practitioners about patients they have in common enhances total patient care, but such communication rarely occurs. This may be due to lack of appreciation by doctors of the medical risks to certain patients undergoing dental treatment. To ascertain a relevant medical history, prospective medical screening was performed on 1500 new patients attending a general dental practice using a standard health questionnaire followed by an interview between the patient and dentist. There were 382 (25.5%) patients with a current or past medical history of relevance to dentistry, 90 (6.0%) were taking medication of potential importance and 105 (7.0%) considered they had an intolerance to certain drugs. The screening provided a patient data base for medical and medico-legal purposes. A total of 376 (25.1%) questionnaires were filled out incorrectly and 63 of these (16.8%) had major misinformation about medical history. A small but important group deliberately misled the dentist either from fear of refusal of treatment or embarrassment about their medical history. Therefore interviews are an essential adjunct to written health questionnaires in eliciting accurate information. Formal screening of new patients is essential in general dental practice. Furthermore, general medical practitioners need to become aware of the common risks to patients undergoing dentistry. Better formal and informal communication between general medical and dental practitioners is recommended for the benefit of their mutual patients. PMID- 2560024 TI - An assessment of the value of video recordings of receptionists. AB - Video recordings of receptionists at work in general practice were found to be useful for self assessment by the receptionists and enabled the doctors to see areas for improvement in the organization of the reception area. PMID- 2560023 TI - Sexually transmitted diseases and Chlamydia trachomatis in women consulting for contraception. AB - To study the frequency of genital infections in women consulting their family doctor for contraception, 248 women (median age 23 years) were examined for a range of genital microorganisms. The prevalence of clue cells, Candida albicans and Trichomonas vaginalis were 21.0%, 12.9% and 2.0%, respectively. Neisseria gonorrhoeae was isolated in only one case, whereas Chlamydia trachomatis was found in 6.3% of women. A specific clinical picture for an infection with C. trachomatis in women was not seen. Given the prevalence of over 5% for C. trachomatis and the absence of typical signs and symptoms in infected women, screening for this organism is recommended in women requesting an intrauterine contraceptive device, to prevent complications such as pelvic inflammatory disease and their sequelae. PMID- 2560025 TI - Facilitating a trainee collaborative study. AB - The Essex faculty of the Royal College of General Practitioners organized a collaborative study for trainees in Essex between October 1986 and July 1988. Of the trainees in post during the study period, 28 (46%) participated. The study was performed not only as an educational exercise for trainees in their practice year but also to assess the feasibility of collaborative study as a research tool in general practice. The authors feel that facilitating collaborative research is a faculty activity worthy of consideration. PMID- 2560027 TI - Compulsory audit projects for medical students. PMID- 2560026 TI - Traveller gypsies and primary care. AB - Traveller gypsies have resided in the British Isles for over 500 years, making them one of our oldest ethnic minorities. They experience widespread prejudice and discrimination from the settled population. In the sphere of health care the marginalization of traveller gypsies has resulted in poor access to services and relative neglect of their health needs. In this paper the health of traveller gypsies is reviewed from the perspective of primary care, and the role of general practitioners in improving health care for this community is discussed. PMID- 2560029 TI - Effect of small group education on the outcome of chronic asthma. PMID- 2560028 TI - Trainee exchanges. PMID- 2560030 TI - Comparison of the workload of a trainer and trainee. PMID- 2560032 TI - Patient satisfaction. PMID- 2560031 TI - Continuing medical education. PMID- 2560034 TI - Dispensing costs. PMID- 2560033 TI - Prescribing research: PACT to the future. PMID- 2560035 TI - Community hospitals. PMID- 2560036 TI - Compulsory admission to hospital. PMID- 2560037 TI - Needs of elderly people in residential homes. PMID- 2560038 TI - Can general practitioners counsel? PMID- 2560039 TI - Fellowship by assessment. PMID- 2560040 TI - Staying the distance. PMID- 2560041 TI - The future role of mental illness hospitals. PMID- 2560042 TI - Organized personal care--an effective choice for managing diabetes in general practice. AB - A system of diabetic review was introduced in two Southampton training practices in March 1985. Each partner, with the help of the practice nurse, retained responsibility for review of their own diabetic patients. During the study period (1984-86) 213 diabetics remained with the practices. In 1984 there were 94 non insulin dependent patients who were not receiving hospital outpatient care. Over the study period there was an increase in the surveillance of blood glucose, blood pressure, weight, urine (for protein), fundi, visual acuity and feet for this group so that in 1986 between 79% and 89% of patients were having these parameters checked at least annually. More complications were found and more referrals for specialist evaluation were made. There was a trend towards transfer of care from the hospital to the general practitioner, and the proportion of non insulin dependent diabetic patients receiving their care entirely from general practice increased from 22% to 60% over the period. There was a small increase in the workload of the general practitioners and a considerable contribution to care was made by the practice nurses. It is concluded that structured personal diabetic care based on a nurse coordinated service is a satisfactory alternative to the 'specialist' general practitioner mini-clinic model. PMID- 2560043 TI - Ethical decision making by British general practitioners. AB - General practitioners in England and Wales were sent a questionnaire asking how they would handle the ethical problems posed by six case vignettes and their reasons for their decisions. The ethical problems included: how much information to divulge to patients, how extensively a physician should become involved in the lifestyles of patients and how to deal with a possible family problem. The varying patterns of response to the six cases suggested that ethical issues are resolved in a case-by-case, not a theoretical, basis. PMID- 2560044 TI - Patients' satisfaction with general practitioner services: a survey by a community health council. AB - Brighton community health council responded to the invitation of five local general practitioners to undertake a patient satisfaction survey of their practices. A total of 177 mothers of children under five years old were interviewed in their own homes. Satisfaction with the services provided by the general practitioners and members of the primary health care team to the respondents and to their young children was high overall, but critical comment reflected a dissatisfaction with professionals' unwillingness to take mothers' concerns at face value and to recognize the validity of mothers' own experiential knowledge. Some women were not satisfied with the extent to which they could ask questions or explain their problem. They resented attempts by receptionists to bar access to doctors and the apparent reluctance of doctors and health visitors to make home visits. It is suggested that various strategies such as telephone consultations, written guidelines on childhood ailments and parent support groups within the context of a more interactive partnership between patients and professionals could lead to a more effective service. PMID- 2560045 TI - Women's experiences of general practitioner management of miscarriage. AB - A study of 67 women one month after miscarriage identified significant levels of dissatisfaction with their medical care. There are particular problems in managing miscarriage which is very distressing for many women but a common clinical presentation for doctors. The reasons for women's dissatisfaction with their management are explained. Greater understanding of the experience of miscarriage should lead to better management and suggestions are made for better care for this common distressing experience. PMID- 2560047 TI - Should performance indicators in general practice relate to whole practices or to individual doctors? AB - In a study of referrals to East Anglian hospitals 737 referrals in six specialties from three general practices were examined to see how accurately the hospital computer master index had identified the referring practice, the referring general practitioner and the doctor with whom the patient was registered. Although the practice was accurately identified by the hospital computer in 97% of referrals, the identification of the referring doctor and the patient's registered general practitioner were less reliable (72% and 49% respectively). It is concluded that at present the practice rather than the individual doctor may be the appropriate unit of analysis for studies of general practitioners' referral rates. This may be true for other performance indicators where information on a doctor's case mix and workload is not available. The results of this study emphasize that problems may arise if data relating to individual general practitioners are interpreted out of context of the practice and the way in which it is organized. PMID- 2560046 TI - Measles--mumps--rubella immunization: the role of the general practitioner in achieving a high uptake. AB - A survey of all general practitioners in Fife conducted prior to the introduction of measles-mumps-rubella immunization on a pilot basis in May 1987 showed that 85% considered mumps worth preventing and 94% believed the rubella programme worth augmenting with universal childhood immunization. Ninety seven per cent considered measles worth preventing and 98% were prepared to recommend measles mumps-rubella immunization to parents instead of measles vaccine. One year after introducing the measles-mumps-rubella vaccine in Fife, 91% of children had been immunized with the combined vaccine or measles vaccine before their second birthday. This compares with the 83% that received measles vaccine before the combined vaccine was introduced. Eighty per cent of preschool children were also immunized with the combined vaccine at school entry in a catch-up exercise. This study demonstrates that there are few major professional barriers to achieving a high uptake of the measles-mumps-rubella vaccine in this area. The vaccine was introduced nationally on 1 October 1988 and its uptake is likely to exceed the current unsatisfactory level achieved with measles vaccine. However, this outcome will largely depend on the commitment of doctors to the programme. PMID- 2560048 TI - Standardization of core data for practice annual reports: a pilot study. AB - To investigate the feasibility of a standardized practice annual report, nine general medical practices in Grampian region collected data over a four month trial period. The definitions for core data used were arrived at after considerable consultation and discussion. They were found to be workable indicators of practice activity enabling comparisons to be made between practices. Three practices reported that they were completing forms more accurately and that this had led to increased item of service income. The results from this study show smaller variations than have been previously reported and there is a case to be made for the pooling and collation of such defined core data. Analysis of this practice activity would provide the stimulus for further research. PMID- 2560049 TI - The practice annual report: post mortem or prescription? AB - While a minority of general practices have been producing annual reports for a number of years one of the implications of the government's new contract is that this process will become much more widespread. To date the majority of reports have concentrated on looking back at past activity rather than on looking forward. In this article a mechanism is outlined for turning the annual report into a vehicle for improving participation; for enabling performance review; and for providing direction for the future. Central to this process is the incorporation of a number of objectives into the report. Objectives should relate to the resources of the practice, the activities of the practice and the results which the practice hopes to achieve. Methods for deriving appropriate objectives are described. PMID- 2560051 TI - Foreign body inhalation: a danger of metered dose inhalers. PMID- 2560052 TI - Patients' resistance to change. PMID- 2560050 TI - Down's syndrome and the general practitioner. AB - People with Down's syndrome form a heterogeneous group sharing a single constant feature--an extra chromosome. This paper reviews the many clinical problems associated with Down's syndrome and emphasizes the prevention of secondary handicapping conditions. Current policies on antenatal screening for Down's syndrome are discussed. The review draws attention to the need for general practitioners to see themselves as part of a network of community services providing support to people with Down's syndrome and their families. PMID- 2560053 TI - Spiritual healing in general practice. PMID- 2560054 TI - Quality assessment or quality control? PMID- 2560056 TI - Otitis media with effusion. PMID- 2560055 TI - Direct access to hospital investigative facilities. PMID- 2560057 TI - Effect of small group education on the outcome of chronic asthma. PMID- 2560058 TI - Future of general practice postgraduate education. PMID- 2560059 TI - Trainer and trainee workloads. PMID- 2560060 TI - Patient satisfaction. PMID- 2560061 TI - Changing responsiveness of luteal cells of the marmoset monkey (Callithrix jacchus) to luteotrophic and luteolytic agents during normal and conception cycles. AB - Dispersed marmoset luteal cells were incubated for 2 h and progesterone production measured after exposure to hCG, cloprostenol, dibutyryl cAMP, PGF-2 alpha, PGF-2, adrenaline or melatonin. The cells were studied on Days 6, 14 and 20 after ovulation in conception and non-conception cycles. Luteal cells from Day 14 non-pregnant marmosets were compared with human luteal cells taken in the mid luteal phase. All the treatments stimulated progesterone production including cloprostenol, which is luteolytic when administered to the marmoset in vivo, but the degree of response varied with the stage of the cycle or pregnancy and between marmoset and human luteal cells. In the marmoset, overall analysis of the effect of the treatments showed that, on Day 6 after ovulation, there was no significant effect of any of the treatments in cells from pregnant or non pregnant animals. In contrast, luteal cells from non-pregnant animals on Day 14 showed a significant response to the treatments (F (8,41) = 2.79, P less than 0.0145) whereas cells from pregnant Day-14 animals were responsive; in cells from pregnant animals, the control production of progesterone was high and already equivalent to the levels stimulated by the treatments. By Day 20, cells from pregnant animals produced lower control concentrations of progesterone than did those on Day 14 and there was a significant overall effect of the treatments (F (8,33) = 3.78, P less than 0.003). These results show that the marmoset CL gains responsiveness to treatment between Days 6 and 14 after ovulation in the non pregnant cycle. In pregnancy, on Day 14, 2 days after attachment of the embryo, the high control concentrations of progesterone and absence of response to treatment suggest that an embryo message may have affected the CL, providing an endogenous stimulus. PMID- 2560062 TI - Plasma concentrations of 15-keto-13,14-dihydro-PGF-2 alpha, oestrone sulphate, oestradiol-17 beta and progesterone in pregnant guinea-pigs treated with polychlorinated biphenyls. AB - Guinea-pigs treated by gavage with a total dose of 100 mg polychlorinated biphenyls (PCB: Clophen A50) during Days 17-61 of gestation had higher plasma concentrations of 15-keto-13,14-dihydroprostaglandin F-2 alpha, oestrone sulphate and oestradiol-17 beta during the later stages of gestation than did vehicle treated guinea-pigs. No changes were observed in plasma progesterone concentrations. Our results provide no support for the hypothesis that an enzyme induced decrease in progesterone concentrations is the main cause of the fetal death observed in PCB-treated guinea-pigs. PMID- 2560063 TI - Ca2+ pumping ATPase of cardiac sarcolemma is insensitive to membrane potential produced by K+ and Cl- gradients but requires a source of counter-transportable H+. AB - The sensitivity of the Ca2+ pumping ATPase of bovine cardiac sarcolemma (SL) to changes in membrane potential was studied in a preparation of sealed SL vesicles. Membrane potential was imposed by preincubating the vesicles in media of defined ion composition (K+, Cl-, choline+ and gluconate-) and diluting into media of differing ion composition. The durations of the ion gradients and relative ion permeabilities were determined in separate experiments by the dependence of the half time for net K+ (or choline+) movement coupled with these anions (Cl- or gluconate-), registered by the fluorescence of 1-anilino-8-naphthalene sulfonate (Chiu, V.C.K., Haynes, D.H. 1980. J. Membrane Biol. 56:203-218). Relative permeabilities were: 1.0, K+; greater than or equal to 10.0, 1 microM valinomycin K+; 4.0, Cl-; 0.66, choline+; 0.38, gluconate-. Durations of the gradients ranged between 17 sec (KCl, valinomycin) to 195 sec (K(+)-gluconate-). In separate experiments, active Ca2+ uptake was monitored using chlorotetracycline (CTC) fluorescence, a technique validated by 45-Ca2+ measurements (Dixon, D., Brandt, N., Haynes, D.H. 1984. J. Biol. Chem. 259:13737-13741). Active Ca2+ uptake was initiated in the presence of monovalent ion gradients. The values of the membrane potentials (Em) imposed by the monovalent ion gradients were calculated using the ion concentrations, their relative permeabilities and the Goldman-Hodgkin-Katz equation. No effect of membrane potential on transport rate was observed (less than or equal to 4%, for 5-7% SD) for imposed potentials as extreme as greater than or equal to +71 and less than or equal to -67 mV. Formal analysis shows that the above observations are not compatible with models in which the Ca2+ pumping ATPase functions in an electrogenic or charge-uncompensated fashion. Further experimentation showed that the pump rate is slowed when uptake is measured at less-than-adequate concentrations of buffer (5 vs. 25 mM HEPES/Tris). This, together with further control experiments using nigericin and FCCP, gave evidence that the pump requires a source of counter-transportable H+ in the vesicle lumen. The above experimentation also underlines the need for control of internal pH to obviate erroneous interpretation of ion perturbation experiments. The results are compared with results obtained with the Ca2+ ATPase pump of skeletal sarcoplasmic reticulum. PMID- 2560064 TI - Human erythrocytes have binding sites for beta-endorphin. AB - Monoiodinated human beta-endorphin was found to bind specifically to human erythrocytes. Unlabeled beta-endorphin and beta-endorphin inhibited binding, but (-)naloxone, [D-Ala2, D-Leu5]-enkephalin, and leu- and met-enkephalin did not. Immunoelectron microscopy, using rabbit anti-beta-endorphin antibody, an antirabbit IgG secondary antibody, and complexed horseradish peroxidase, revealed that at low concentrations beta-endorphin binds to the cell surface. Electron spin resonance spectroscopy showed no effect of beta-endorphin on membrane fluidity. This receptor does not appear to conform to the characteristics of an opiate receptor. PMID- 2560065 TI - [Clinical significance of serum ACTH analysis in pituitary function tests]. PMID- 2560066 TI - [Clinical significance of urinary tetrahydro-11-deoxycortisol analysis in endocrine tests of the adrenal cortex]. PMID- 2560068 TI - [Clinical significance of serum and urinary 11-hydroxycorticosteroid analysis in endocrine tests of the adrenal cortex]. PMID- 2560067 TI - [Clinical significance of serum 11-deoxycorticosterone analysis in endocrine tests of the adrenal cortex]. PMID- 2560069 TI - [Clinical significance of urinary 17-hydroxycorticoid and its component analysis in endocrine tests of the adrenal cortex]. PMID- 2560070 TI - [Clinical significance of serum and urinary 18-hydroxycorticosterone analysis in endocrine tests of the adrenal cortex]. PMID- 2560071 TI - [Clinical significance of urinary 17-ketogenic steroids and its component analysis in endocrine tests of the adrenal cortex]. PMID- 2560073 TI - [Clinical significance of catechol estrogen analysis in endocrine tests of the gonadal and placental system]. PMID- 2560072 TI - [Clinical significance of urinary 17-ketosteroids and 17-KS fraction analysis in endocrine tests of the adrenal cortex]. PMID- 2560074 TI - [Clinical significance of pregnancy-specific beta 1-glycoprotein analysis in endocrine tests of the gonadal and placental system]. PMID- 2560075 TI - [Clinical significance of HCG and subunit analysis in endocrine tests of the gonadal and placental system]. PMID- 2560076 TI - [Clinical significance of kininogen-proteinase complex analysis in biochemical laboratory tests]. PMID- 2560077 TI - [Clinical significance of collagenase analysis in biochemical laboratory tests]. PMID- 2560078 TI - [Significance of 5'-nucleotidase analysis in clinical tests]. PMID- 2560079 TI - [Significance of prolyl hydroxylase analysis in clinical tests]. PMID- 2560080 TI - [Determination of oxypurine and its clinical implication]. PMID- 2560081 TI - [Clinical significance of thrombomodulin analysis in blood coagulo-fibrinolytic tests]. PMID- 2560082 TI - [MRI imaging of liver neoplasma]. AB - MRI by 1.5 T imager with spin echo pulse sequence was performed on 53 liver neoplasms (13 hepatocellular carcinomas (HCC), 28 cavernous hemangiomas and 12 metastasis). Lobulated and notched contour were observed in 72% of our cases of hemangioma. It is well known that in hemangioma several kinds of echographic patterns appear, but in MRI, almost all hemangioma appear to show the same signal pattern as high intensity homogeneous on T2 weighted image. No liver neoplasma showed high intensity on T1 image except in 1/3 of cases of HCC. Contrast enhancement with Gd-DTPA is useful for differential diagnosis of liver neoplasma because it shows different signal patterns from the other diseases. PMID- 2560083 TI - [A case of icteric hepatoma which succeeded internal fistulation after radiotherapy]. AB - A case of icteric hepatoma was treated by TAE, arterial chemoinfusion and photoirradiation to the hepatic hilum. The patient was successfully discharged after radiotherapy, which was most effective for internal fistulation of obstructed bile duct and decreasing the size of portal tumor thrombus. PMID- 2560084 TI - [A long surviving hepatocellular carcinoma case (2 years 1 month) with tumor thrombus in the portal trunk treated by embolization]. AB - A 55-year-old male patient suffered from hepatocellular carcinoma (HCC) with tumor thrombus in the portal trunk and cavernous transformation in the liver hilus. He was treated 4 times effectively by transcatheter arterial embolization (TAE). Tumor thrombus in the portal trunk which was demonstrated by portography before TAE disappeared following initial TAE. He is still alive 2 years and 1 month after initial TAE. We reported a long surviving case with advanced HCC treated by TAE and TAE for advanced HCC was discussed. PMID- 2560085 TI - [A case of multilocular cystic nephroma]. AB - Multilocular cystic nephroma is an uncommon and usually benign nephroblastic neoplasm. Considering its therapeutic indication, it is important to distinguish this lesion from malignant renal masses. The lesion usually shows a well encapsulated, multilocular cystic mass on US and CT. Occasionally, imaging studies are not useful for preoperative diagnosis, however, surgical biopsy should be considered for correct pathological diagnosis. PMID- 2560087 TI - [Renal function and humoral factors]. PMID- 2560086 TI - Distribution of varicella-zoster virus strains carrying a PstI-site-less mutation in Japan and DNA change responsible for the mutation. AB - We have analyzed 53 varicella-zoster virus (VZV) strains isolated in Japan for the presence of a PstI cleavage site in a middle portion of the long unique region of VZV genome. About 25% of the strains examined did not have this PstI cleavage site, although the majority of strains retained this site. For several VZV strains lacking this PstI cleavage site, we sequenced a recombinant DNA derived short segment containing the mutation. In all strains analyzed, we identified a base replacement from A to G within the PstI recognition sequence. Because no wild-type VZV strain carrying this mutation has been found so far in other countries, it is very likely that the mutation once occurred in the past and has been conserved in some strains in Japan. We have concluded that the PstI site-less mutation is reliable marker for discrimination of VZV strains in Japan. PMID- 2560088 TI - [Noninvasive diagnosis of cardiac involvement by technetium-99m-pyrophosphate (Tc 99m PYP) myocardial scintigraphy in 2 cases of familial amyloid polyneuropathy and 1 case of secondary amyloidosis]. AB - To validate the significance of technetium-99m-pyrophosphate (Tc-99m PYP) myocardial scintigraphy in diagnosing cardiac amyloidosis, 2 patients with familial amyloid polyneuropathy (FAP) and 1 patient with amyloidosis secondary to chronic rheumatic arthritis were studied. All three patients had echocardiographic abnormalities, which were increased wall thickness of the interventricular septum and the left ventricular posterior wall, and granular sparkling appearance in the septum. In 2 patients with FAP, abnormal myocardial uptake of Tc-99m PYP was diffusely detected in Tc-99m PYP SPECT. In the remaining 1 patient with secondary amyloidosis, however, Tc-99m PYP SPECT showed no abnormality, although we had confirmed the presence of myocardial amyloid deposits (type AA amyloid protein) with high amount in the histological examination. Thus, these results indicate that Tc-99m PYP scintigraphy may have a limitation in detecting cardiac involvement in secondary amyloidosis although it is useful in FAP. PMID- 2560089 TI - [Lung perfusion scintigraphy by SPECT--new application of Bull's eye analysis]. AB - The present study reports the characteristic performance using lung segmental phantom filled in Tc-99m-pertechnetate. To evaluate the segmental defect in lung perfusion scintigraphy, we applied Bull's eye analysis in addition to planar image set. Bull's eye analysis especially facilitated the interpretation in both middle and lower lobes. PMID- 2560090 TI - [The analysis of Tc-99m RBCs time activity curve on liver tumor--differentiation between hemangioma and hepatocellular carcinoma]. PMID- 2560091 TI - Diazepam potentiates the corticoidogenic response of bovine adrenal fasciculata cells to dibutyryl cyclic AMP. AB - To provide a possible role of peripheral type benzodiazepine receptors in the regulation of glucocorticoid biosynthesis. We have examined the effect of diazepam on the corticoidogenic response to dibutyryl cyclic AMP in isolated bovine adrenal fasciculata cells. Diazepam alone (up to 100 microM) had no effect on the corticoidogenesis. Diazepam caused a dose-dependent potentiation of dibutyryl cyclic AMP-induced corticoidogenesis. However, diazepam had no effect on the corticoidogenic response to ACTH and a high concentration of KCl. The potentiating effect by diazepam was clearly detected after 90 min-incubation, and it was blocked by YM-684 (diazepam antagonist) and ML-236B (cholesterol de novo synthesis inhibitor). Diazepam caused no significant decrease of intracellular content of cholesteryl esters during the corticoidogenic response to dibutyryl cyclic AMP. When the cells were incubated in the presence of (+)-PN200-110, a potent voltage-dependent Ca channel inhibitor, the potentiating effect by diazepam was not affected in spite of a significant inhibition of dibutyryl cyclic AMP-induced corticoidogenesis. These results indicate that the potentiating effect of diazepam on dibutyryl cyclic AMP-induced corticoidogenesis is due in part to the activation of the intracellular cholesterol supply system (cholesterol de novo synthesis) without any change of voltage-dependent Ca channels. PMID- 2560092 TI - Characterization of calcitonin gene-related peptide (CGRP) receptors in guinea pig lung. AB - Receptors for calcitonin gene-related peptide (CGRP) in the lung membranes of guinea pig were characterized by using a millititer plate precoated with polyethylenimine. Specific binding of 125I-CGRP was time-dependent, rapid, and reversible, and the binding increased linearly with increasing concentrations of membrane protein. Scatchard analysis revealed two classes of CGRP binding sites: high affinity sites with a KD value of 7.17 x 10(-11) M and a Bmax of 364 fmol/mg protein and low affinity sites with a KD value of 1.7 x 10(-8) M and a Bmax of 39594 fmol/mg protein. Furthermore, the specific binding of 125I-CGRP was dissociated in the presence of GTP or Gpp(NH)p, suggesting that CGRP receptors in guinea pig lung membranes were coupled to the guanine nucleotide regulatory protein. Scatchard analysis of CGRP binding in the presence of GTP revealed selective inhibition of the binding to high affinity binding sites. Unlabeled CGRP displaced the binding of 125I-CGRP to guinea pig lung membranes with an IC50 value of 3.1 x 10(-10) M. In contrast, salmon calcitonin and human calcitonin displaced the binding at 600-fold higher concentrations. We suggest that both low and high affinity binding sites for CGRP exist in the lung membranes of the guinea pig, and the high affinity binding sites for CGRP may be coupled to GTP binding regulatory protein. PMID- 2560093 TI - Blockade of the development of analgesic tolerance to morphine by psychological stress through benzodiazepine receptor mediated mechanism. AB - beta-Carboline-3-carboxylic acid ethyl ester (beta-CCE) dose-dependently potentiated psychological-stress induced analgesia (PSY-SIA), and the effect was reversed by diazepam. Concurrent exposure to PSY stress or concomitant treatment with beta-CCE blocked the development of analgesic tolerance to morphine; the effect of PSY stress was antagonized by diazepam, and that of beta-CCE was reversed by Ro 15-1788. These results suggest that psychological factors which are mediated through benzodiazepine receptors are involved in the mechanism for blocking the development of analgesic tolerance to morphine by PSY stress. PMID- 2560094 TI - Ionic currents in a mammalian intestinal smooth muscle--from tissue to single channels. PMID- 2560096 TI - [Functional scintigraphy of radioactive cardiac blood pool: amplitude and phase analysis and conventional parameters of left-ventricular wall motion at rest]. AB - Authors performed quantitative assessment of 13 global and regional, left ventricular wall motion parameters obtained from 52 radioactive blood pool cardiac scintigrams. Of these performed in 1986 examinations, authors selected 3 equipotent patients' groups (myocardial infarction, cardiac aneurysm, cardiomyopathy) thus the contractility disorders were located in inferior wall segments. 13 examinations of the same material, without contractility disorders were selected (standard). Using time-activity curve authors measured following global parameters: ejection fraction (EFg1), mean ejection rate (MER), peak ejection rate (PER), peak filling rate (PFR), time from the end-systole to peak filling rate (tPFR) and regional parameters such as: regional and local ejection fraction (EFr and EF1), time from the beginning of the cycle to the end-diastole (ED) and to the end-systole (ES), contractility (KURCZ), amplitude of contractions. (AMPL) and of phase (PHASE) as well as standardized variables of amplitude distribution (SDU). The most significant quantitative parameter was EFgl. Dyskinesis symptom was the most important for cardiac eneurysm diagnosis. Phase shift served a confirmation of aneurysm earlier diagnosed using a cinematic display. PMID- 2560095 TI - Contribution of peripheral opioid receptors to the trimebutine-induced contractions of the proximal colon in anesthetized rats. AB - In this study we investigated the involvement of opioid receptors in the contractile response to trimebutine using with the proximal colon of anesthetized rats. Trimebutine (3 mg/kg i.v.) enhanced spontaneous contractions of the proximal colon in anesthetized rats. The contractile response was partially inhibited by intravenous administration of an opioid antagonist, naloxone at 1 approximately 30 micrograms/kg, but was hardly depressed by intracisternal administration of naloxone (30 micrograms/kg). Morphine (30 micrograms/kg i.v.) evoked colonic contractions which were abolished by intravenous naloxone (30 micrograms/kg). These results suggest that the colonic contractions evoked by trimebutine in anesthetized rats are, in part, mediated by peripheral opioid receptors. PMID- 2560097 TI - Limited proteolysis and differential down-regulation of protein kinase C subspecies in KM3 cells. AB - Protein kinase C (PKC) is known to play crucial roles in signal transduction. This enzyme was originally found as an undefined protein kinase, which could be activated by limited proteolysis by a Ca2(+)-dependent neutral protease, calpain. PKC was thought to be a single entity for many years, but recent, molecular cloning and biochemical analysis has shown that this enzyme consists of multiple subspecies with closely related structures. The present studies were undertaken to explore the susceptibility of PKC subspecies to proteolysis by calpain in KM3 cells, a pre-B, pre-T cell line. The PKC from KM3 cells was resolved into two subspecies, type II (mainly beta II) and type III (alpha), upon hydroxyapatite column chromatography. This cell line was also shown to contain calpain I and calpain II, which show different sensitivities to Ca2+. Both type II and type III PKC were hydrolysed by either calpain I or calpain II, but type II PKC was cleaved more effectively than type III. The simultaneous presence of phospholipid and diacylglycerol enhanced the cleavage of these PKC subspecies by calpain I. When these cells were treated with phorbol ester, the rates of translocation and down-regulation of these PKC subspecies were different; type II PKC was translocated and depleted from the cell more rapidly than type III enzyme. The results presented herein suggest different roles of the PKC subspecies in the cellular responses to external stimuli. PMID- 2560098 TI - Copper deficiency in the mitochondria of cultured skin fibroblasts from patients with Menkes syndrome. AB - The mitochondrial copper concentrations and cytochrome C oxidase activity of the fibroblasts from the patients with Menkes syndrome were investigated. Both the mitochondrial copper concentrations and cytochrome C oxidase activity of fibroblasts from patients with Menkes syndrome were lower than those of the control fibroblasts. These data indicate that the mitochondria of fibroblasts from patients with Menkes syndrome are in a state of copper deficiency. The activity decline of cytochrome C oxidase, a mitochondrial cuproenzyme, seems to be caused by copper deficiency in the mitochondria. PMID- 2560100 TI - [Syncope and transient coronary ischemia after the first dose of enalapril]. PMID- 2560099 TI - Further analysis of the disturbed adrenocortical function in the cerebro-hepato renal syndrome of Zellweger. AB - Plasma aldosterone, corticosterone, 11-deoxycorticosterone, progesterone, 17 hydroxyprogesterone, 11-desoxycortisol, cortisol and cortisone were determined simultaneously in small plasma samples obtained during an i.v. ACTH (Synacthen) stimulation test in five patients (age 4-7 years) with Zellweger syndrome. The response of all ACTH-dependent steroids to Synacthen was severely impaired in all patients, despite normal basal levels. It can be concluded that the biochemical defect in adrenal steroidogenesis causing the inadequate response to ACTH injection is located proximal to progesterone. We propose that the lack of responsiveness to ACTH is secondary to an abnormality of ACTH receptors on the adrenocortical cell. The extremely low levels of non-specific lipid transfer protein may be a contributing factor. PMID- 2560101 TI - [What is the disease course in carriers of HBsAg?]. PMID- 2560102 TI - [Gastric ulcer as a unique and symptomatic manifestation of infection by cytomegalovirus in a healthy adult]. AB - A previously healthy female without risk factors had cytomegalic inclusion disease with symptomatic gastric involvement and without evidence of systemic cytomegalovirus (CMV) infection. Multiple ulcerations with a fibrinous bottom were discovered by fibrogastroscopy in the gastric body and antrum. Histological study showed cells with intranuclear inclusions and basophilic nuclear cytoplasm. The detection of specific IgM and its subsequent negativization suggest an active CMV infection. Gastric infection may be the isolated and symptomatic feature of the disease in the healthy adult. PMID- 2560103 TI - Dimeric dermorphin peptides: central administration suppresses gastric acid secretion through interaction with mu-type opioid receptor. AB - The effect of centrally administered dermorphin and a series of dimeric dermorphin analogs on gastric acid secretion was studied in pylorusligated rats. Dimeric dermorphin analogs consist of identical peptide chains either directly linked with a dihydride bond or a polyethyleneamine bridge at their carboxy termini. At a dose of 0.5 nmol dermorphin, and the dimeric analogs di-[D Arg2,Sar4]-tetra-DM2, di-penta-DM0, di-[Sar4]-penta-DM0, di-[D-Arg2,Sar4]-penta DM0, di-[D-Ala4]-penta-DM0 and di-DM0, and di-DM0, inhibited gastric acid output in a statistically significant manner (P less than 0.05). Furthermore, the binding characteristics of these peptides for the mu-type opioid receptor were analyzed using a brain synaptosomal fraction. Dermorphin and several dimeric dermorphin peptides bound to the mu-receptor: di-penta-DM0, di-[Sar4]-penta-DM0, di-penta-DM2 and di-DM0, had 3-to 5-fold greater affinity for the mu-receptor than the specific mu-agonist DAGO. These data indicate that a correlation exists between the central mediated gastric inhibitory effect of DM and several dimeric analogs and their affinity for the mu-type opioid receptor in rat brain. PMID- 2560104 TI - High plasma levels of cortisol in patients with senile dementia of the Alzheimer's type. AB - Plasma cortisol levels and other factors including thyroid hormone in patients with Alzheimer's type (n = 10), vascular type (n = 10) or mixed type (n = 10) senile dementia were compared with those in non-demented senile controls (n = 10). Plasma cortisol levels at 8:00 a.m. in Alzheimer's type dementia and mixed type dementia were 17.3 +/- 4.3 micrograms/dl (mean +/- SD) and 15.6 +/- 2.3 micrograms/dl, respectively. These values were significantly higher (p less than 0.005 and p less than 0.01) than those found in the control subjects (12.0 +/- 3.1 micrograms/dl). Plasma cortisol levels in vascular-type dementia (14.4 +/- 6.3 micrograms/dl) did not differ significantly from those in the controls. Plasma ACTH in senile dementia of the Alzheimer's type was lower, but not significant as compared with that in normal controls. In three subgroups of senile dementia and normal controls, plasma cortisol levels inversely correlated significantly with the degree of cognitive function. Plasma levels in TSH-thyroid system and blood pressure did not show any significant change in three types of senile dementia. These data suggest that senile dementia of the Alzheimer's type accompanies relatively and primarily high plasma cortisol levels and this may associate with cognitive dysfunction in Alzheimer's type senile dementia. PMID- 2560105 TI - [The value of laparoscopy. Results of 600 studies performed in an internal medicine department at the General Hospital of Dakar]. AB - 600 laparoscopic examinations were performed in an Internal Medicine Department of the general Hospital Dakar (Senegal) from 1984 to 1989 in 402 males and 198 females aged from 7 to 82 years. This examination of simple and quick procedure well tolerated under simple premedication, should be performed only after diffusion of the pneumoperitoneum, per- foration being the major risk. Even if liver cell carcinoma and cirrhosis represent more than an half of the examined cases (52%) due to their high frequency, exploration of peritoneal pathology is the best indication for laparoscopy. Thanks to laparoscopy, 68 tuberculosis, 28 peritoneal carcinomatosis and 11 infectious peri-hepatitis were diagnosed. It remains a major examination perfectly adapted to studies of the so-frequent hepato-peritoneal pathology in Black Africa] PMID- 2560106 TI - [Chronic calcifying pancreatitis in hospitals in the Ivory Coast]. AB - The authors report on 34 cases of chronic calcifying pancreatitis in Ivory Coast. Chronic calcifying pancreatitis is rare (0.24 P.C. of admitted patients) and appears mainly in man 40 years old, revealed by diabetes (73.5 P.C.). Principal aetiology was alcoholism, 50 P.C. Genuine tropical chronic calcifying pancreatitis exists in small number. It seems its physiopathological mechanism is not only nutritional. PMID- 2560107 TI - [Analysis of 3 years of blood cultures in the University Hospital Center of Treichville (Abidjan)]. AB - From august 1985 to september 1988, 7819 blood cultures aero-anaerobic bacteria in a Brain Heart-broth. 1383 blood cultures were positive (18%). We isolated first Salmonella, followed then by Staphylococci. PMID- 2560108 TI - Inhibition by the protein kinase inhibitors, isoquinolinesulfonamides, of fluid accumulation induced by Escherichia coli heat-stable enterotoxin, 8-bromo-cGMP and 8-bromo-cAMP in suckling mice. AB - The effects of isoquinolinesulfonamides, which inhibit protein kinase, on fluid accumulations induced by heat-stable enterotoxin of enterotoxigenic Escherichia coli (STh), 8-bromo-cGMP and 8-bromo-cAMP in suckling mice were studied. Both N [2-(methylamino)ethyl]-5-isoquinolinesulfonamide (H-8) and N-(2-aminoethyl)-5 isoquinolinesulfonamide (H-9) inhibited the fluid accumulation induced by STh. Fluid accumulation induced by four mouse units of STh (four-fold the minimum effective dose, 10 ng) was completely inhibited by 0.4 mumol of H-8 or H-9. H-8 and H-9 also inhibited fluid accumulation induced by 8-bromo-cGMP. On the other hand, H-8 and H-9 only partially inhibited fluid accumulation in suckling mice induced by 8-bromo-cAMP, probably because their affinities to cAMP-dependent protein kinase were lower than their affinities to cGMP-dependent protein kinase. From these results, it is concluded that the activation of cGMP-dependent protein kinase by increase in cGMP by ST or by 8-bromo-cGMP, and very probably the activation of cAMP-dependent protein kinase by increase in cAMP by cholera enterotoxin and heat-labile enterotoxin of enterotoxigenic E. coli and by 8-bromo cAMP are necessary steps in signal transduction following increases in concentrations of cGMP and cAMP in intestinal brush border cells. PMID- 2560109 TI - Oxidative and non-oxidative intracellular killing of Mycobacterium avium complex. AB - Among mycobacteria, those belonging to the Mycobacterium avium complex (MAC) are the most common cause of bacteremia in AIDS patients. To understand better the mechanisms by which human macrophages kill intracellular MAC, we studied in an in vitro test system transparent morphotypes of the three most common bacteremic serotypes from AIDS patients and an opaque variant, obtained in vitro from the most mouse-virulent strain (MAC 101). The three serotypes differed in susceptibility to oxidative bactericidal mechanisms of macrophages. The transparent morphotype of strain 101 (serotype 1) was completely resistant to the intracellular killing effects of a phagocyte's reactive oxygen radicals and hydrogen peroxide, whereas strains 109 (serotype 4), 100 (serotype 8), and the opaque variant from strain 101 were killed by oxidative bactericidal mechanisms. However, even for these bacteria, non-oxidative mechanisms appear to have a role in intracellular killing. PMID- 2560110 TI - Persistent parvovirus B19 infections in humans. PMID- 2560112 TI - Comparative pathogenesis of three strains of pseudorabies virus in pigs. AB - Three strains of pseudorabies virus were intranasally inoculated into 10-week-old pigs and the pathogenesis of the infection was compared. Virulent NIA-3 virus caused widespread necrotic lesions in nasal mucosa, rapidly invading the stroma and infecting axons of olfactory nerves within 24 h of inoculation. Intermediate virulent virus 2.4N3A, a mutant strain derived from NIA-3, caused less necrosis of the mucosa and did not reach axons of olfactory nerves until 72 h after inoculation. Bartha virus strain K, a non-virulent virus strain, caused a mild infection in the superficial layers of nasal epithelium. Viral antigens were not detected in stromal fibroblasts or nerve cells. The inflammatory response of the pigs varied with the virus strains used: after infection with NIA-3 virus mainly neutrophils infiltrated the nasal mucosa, whereas after infection with 2.4N3A virus and Bartha virus, mainly macrophages and lymphocytes infiltrated the nasal mucosa. PMID- 2560111 TI - Effect of Salmonella typhimurium porins on biological activities of human polymorphonuclear leukocytes. AB - The effect of Salmonella typhimurium porins on human polymorphonuclear leukocytes (PMNs) was studied. Labeled porins were shown to bind to the PMNs, and could be completely displaced by unlabeled porins. The binding caused modifications of membrane integrity and of the physico-chemical characteristics of the PMN surface, e.g. decreased oxidative burst, decreased hydrophobicity and altered cell morphology. The porins acted as both chemotaxins and chemotaxinogens. When PMNs were preincubated with porins their migration in the presence of commonly used chemoattractants (serum activated by zymosan or N-formyl-L-methionyl-L leucyl-L-phenylalanine) was inhibited. PMID- 2560113 TI - Molecular cloning of the complete genome of Theiler's virus, strain DA, and production of infectious transcripts. AB - We constructed a complete cDNA clone of the genome of Theiler's virus strain DA in a Bluescript plasmid. This recombinant plasmid, called pTMDA, was used to synthesize full length RNA transcripts of the viral insert. The RNA was infectious for BHK cells. Virus R1-DA, obtained from transfected BHK cells, caused the biphasic disease classically observed with this strain of Theiler's virus. SJL/J mice did not show clinical symptoms during the first week following intracranial inoculation, although viral antigens were found in a few neurons of brain and spinal cord. By 45 days post-inoculation, the mice had developed a chronic demyelinating disease and viral RNA and antigens could be found only in spinal cord white matter in areas surrounded by inflammatory infiltrates. At this stage no RNA or antigens were found in neurons. Therefore the phenotype of R1-DA was indistinguishable from that of genuine DA Theiler's virus. PMID- 2560114 TI - Anti-mouse IgM immunoglobulin protects weanling mice from Coxsackievirus infection. AB - Mice treated from birth with rabbit anti-mouse IgM antiserum (anti-mu), although immunosuppressed in regard to B cell function, may paradoxically be protected when challenged with certain viruses. Our early studies indicated that short term treatment of weanling mice protected against Semliki forest virus, herpes simplex virus, and Coxsackievirus B1. For this study, the murine model of Coxsackievirus B1 infection was chosen to determine if specific anti-mu immunoglobulin (anti-mu Ig) would protect and whether protection could be correlated with antiviral activity in certain organs. Weanling Balb/c mice were treated intraperitoneally (i.p.) with either affinity purified anti-mu Ig or purified normal rabbit (NR) Ig two consecutive days prior to i.p. challenge with Coxsackievirus B1. Mortality of anti-mu Ig treated mice was significantly lower than controls (3.4% vs 89%, P less than 0.001). Importantly, this purified anti-mu-induced protection correlated well with reduced levels of virus in blood and certain organs, especially brain. This decrease was not attributed to interferon (IFN) or virus specific neutralizing (NT) antibody or enhanced cellular cytotoxicity. Anti-mu treatment appears to inhibit virus replication and/or enhance clearance of virus from target organs. The data indicate that a unique antiviral activity is activated by anti-mu treatment, and can be passively transferred to and protect recipient mice. The murine model of Coxsackievirus disease appears to be well suited for use in the elucidation of the mechanism of this protection. PMID- 2560115 TI - Effects of carriage and expression of the Tn10 tetracycline-resistance operon on the fitness of Escherichia coli K12. AB - We have been examining the consequences of alternative modes of regulation of plasmid-borne, Tn10-encoded tetracycline resistance for the fitness of Escherichia coli. In a tetracycline-free environment, we measured the effects on fitness that were caused by (1) maximally induced expression of the resistance operon, (2) low-level constitutive expression of the resistance protein, (3) residual expression of the repressed resistance operon, (4) carriage of the resistance operon, (5) the remainder of the plasmid genome, and (6) hyperexpression of the repressor protein. We observed large reductions in fitness that were associated with induction and with constitutive expression of the tetracycline-resistance protein, but there was no discernible effect of hyperexpression of the repressor protein. We also observed a small reduction in fitness associated with the remainder of the plasmid genome. However, any reductions in fitness that were caused by residual expression and by carriage of the repressed operon were not more than 0.3%. We conclude that tight gene regulation has eliminated antagonistic pleiotropic effects of the resistance gene on fitness, so that possession of an inducible Tn10-encoded tetracycline resistance operon imposes essentially no burden in the absence of antibiotic. PMID- 2560116 TI - Effects of polyoma virus oncogenes in transgenic mice. AB - The effects of polyoma (Py) oncogenes in transgenic mice are reviewed, with emphasis on an analysis of mice carrying individual Py early region genes linked to either the Py early region promoter or to the rat insulin II promoter. The perturbations documented in this series of animals varied with both the Py gene and the promoter. Py promoter-driven transgenes led to the development of pituitary tumors and vascular tumors in a gene-dependent manner, suggesting that the Py T antigens have different tissue specificities of transforming activity. The only pathology found in insulin promoter-driven transgenic mice was a beta cell tumor seen in insulin-promoted Py large T antigen-carrying mice. These results also suggest that the Py oncogenes have different tropisms relative to a specific cell type. Analysis of the Py oncogene-induced perturbations may provide both insights into the molecular events controlling tumorigenesis and access to rare or dispersed cell types for further analysis. PMID- 2560117 TI - Transgenic mouse model of familial amyloidotic polyneuropathy. AB - Familial amyloidotic polyneuropathy (FAP) is a dominantly inherited disorder, characterized by the extracellular deposition of amyloid fibrils composed of variant transthyretin (TTR), and by prominent peripheral nerve involvement. We demonstrate that the main cause of this disease is the presence of a point mutation in the TTR gene. However, neither the time of onset nor the clinical course is predictable. To elucidate the molecular pathogenesis of this disease, we constructed transgenic mice carrying and expressing the human mutant TTR gene. In these mice, amyloid is deposited in the alimentary tract as early as age six months, and becomes more remarkable with aging. These transgenic mice should be useful in elucidating factors which modulate the time of onset and the clinical course of FAP, and in establishing therapy for this intractable disorder. PMID- 2560118 TI - Allelic exclusion and lymphocyte development. Lessons from transgenic mice. AB - The generation of an appropriate, specific immune response to an antigen is a remarkable biological phenomenon. An examination of both allelic exclusion and lymphocyte development is critical for an understanding of this response. Over the last several years, studies using transgenic mice that carry immunoglobulin or T cell receptor transgenes have provided a more detailed understanding of the mechanism of allelic exclusion. Recently, these mice have been used to examine lymphocyte development. In the future, these mice may be used to study the role of lymphocytes in autoimmune diseases. PMID- 2560119 TI - A novel family of potentially mobile DNA elements encoding site-specific gene integration functions: integrons. AB - A family of novel mobile DNA elements is described, examples of which are found at several independent locations and encode a variety of antibiotic resistance genes. The complete elements consist of two conserved segments separated by a segment of variable length and sequence which includes inserted antibiotic resistance genes. The conserved segment located 3' to the inserted resistance genes was sequenced from Tn21 and R46, and the sequences are identical over a region of 2026 bases, which includes the sulphonamide resistance gene sull, and two further open reading frames of unknown function. The complete sequences of both the 3' and 5' conserved regions of the DNA element have been determined. A 59-base sequence element, found at the junctions of inserted DNA sequences and the conserved 3' segment, is also present at this location in the R46 sequence. A copy of one half of this 59-base element is found at the end of the sull gene, suggesting that sull, though part of the conserved region, was also originally inserted into an ancestral element by site-specific integration. Inverted or direct terminal repeats or short target site duplications, both of which are characteristics of class I and class II transposons, are not found at the outer boundaries of the elements described here. Furthermore, the conserved regions do not encode any proteins related to known transposition proteins, except the DNA integrase encoded by the 5' conserved region which is implicated in the gene insertion process. Mobilization of this element has not been observed experimentally; mobility is implied from the identification of the element in at least four independent locations, in Tn21, R46 (IncN), R388 (IncW) and Tn1696. The definitive features of these novel elements are (i) that they include site specific integration functions (the integrase and the insertion site); (ii) that they are able to acquire various gene units and act as an expression cassette by supplying the promoter for the inserted genes. As a consequence of acquiring different inserted genes, the element exists in a variety of forms which differ in the number and nature of the inserted genes. This family of elements appears formally distinct from other known mobile DNA elements and we propose the name DNA integration elements, or integrons. PMID- 2560120 TI - Phase variants of Bordetella bronchiseptica arise by spontaneous deletions in the vir locus. AB - Bordetella bronchiseptica is a common respiratory tract pathogen of many mammalian species. Nucleotide sequences from the locus involved in coordinate regulation of B. pertussis virulence factors, vir, were shown to have a high degree of homology to chromosomal DNA from virulent (Vir+) and avirulent (Vir-) strains of B. bronchiseptica. Small deletions, 50 bp to 500 bp, within the vir locus were found in some of the Vir- phase variants. The vir locus and the adjacent 5' portion of the fhaB structural gene were cloned from the parental Vir+ B. bronchiseptica strain on a 23.5 kb BamHI fragment. Restriction enzyme mapping of the cloned B. bronchiseptica vir locus revealed similarities with and differences from the previously cloned B. pertussis vir locus. The cloned B. bronchiseptica vir locus complemented spontaneous Vir- variants of Bordetella pertussis and B. bronchiseptica as well as vir::Tn5 mutants of B. pertussis. Comparison of various functions of the vir loci of B. bronchiseptica and B. pertussis revealed some interesting differences in the coordinate regulation of virulence factors. PMID- 2560121 TI - Functional analyses of Proteus mirabilis wild-type and mutant RecBCD enzymes in Escherichia coli reveal a new mutant phenotype. AB - Cloned into Escherichia coli the recB, recC and recD genes of Proteus mirabilis produce a recBCD enzyme (exoV) functional in recombination and DNA repair. The direction of transcription of recB, recC and recD, the sizes of the enzyme subunits, and their composition in the active enzyme are similar to that observed for the E. coli enzyme. In lambda crosses, the P. mirabilis enzyme has only about 40% of the Chi activity of the E. coli enzyme. The recBCD genes were also cloned from an exoV mutant of P. mirabilis which is u.v.-sensitive and partly deficient in exoV. The defect was attributed to the recB gene by complementation studies. In a recBCD deletion strain of E. coli, the enzyme from the mutant produced 40% of conjugational recombinants and had retained about 25% of Chi activity. However, it did not restore normal DNA repair, cell viability or recombination in lambda crosses and P1 transduction. The new mutant phenotype is discussed in the light of the assumption that prokaryotic recBCD enzymes can promote recombination in a Chi-dependent and a Chi-independent manner. PMID- 2560122 TI - Preparation of cytidine 5'-monophospho-N-acetylneuraminic acid and uridine 5' diphosphoglucuronic acid; syntheses of alpha-2, 6-sialyllactosamine, alpha-2, 6 sialyllactose, and hyaluronic acid. PMID- 2560123 TI - Production of carbohydrate-specific human monoclonal antibodies in vitro. PMID- 2560124 TI - Phosphomannosyl receptors from bovine testis. PMID- 2560125 TI - High-performance liquid chromatography assays for N acetylglucosaminyltransferases involved in N- and O-glycan synthesis. PMID- 2560126 TI - Mannosidase IA from rat liver Golgi membranes. PMID- 2560127 TI - Glycoprotein processing enzymes of plants. PMID- 2560128 TI - Endo-beta-galactosidase releasing Ga1(alpha 1----3)Gal. PMID- 2560129 TI - Lysosomal alpha-D-mannosidase from rat epididymis. PMID- 2560131 TI - Specificity of Salmonella porin as an eliciting antigen for cell-mediated immunity (CMI) reaction in murine salmonellosis. AB - The specificities of Salmonella porin on elicitations of delayed-type hypersensitivity (DTH) reaction and interleukin-2 (IL-2) production in BALB/c mice immunized with Salmonella typhimurium were examined. Only porin from S. typhimurium was capable of eliciting significant levels of DTH and IL-2 production in S. typhimurium-immunized mice, whereas no significant DTH and IL-2 production were induced by porin from Salmonella enteritidis or Escherichia coli. Our observations suggested that Salmonella porin was a serovar-specific antigen for the elicitation of cell-mediated immunity (CMI) in salmonellosis. PMID- 2560130 TI - Gangliosides that modulate membrane protein function. PMID- 2560132 TI - [The isolation of a concentrated and purified rotavirus preparation by using bentonite]. AB - The preparation obtained when cultivating virus SA-11 in the transplantable cell culture SPEV has been purified and concentrated by means of bentonite. The virus adsorption at acidic values of pH (4.0-4.5) and elution at alkaline values of pH (8.5) were the main stages of production of the concentrated and purified rotavirus preparation. The paper embraces data on the infective activity of the virus protein content, amount of the rotavirus antigen, results of the electron and immunoelectron microscopy, characterizing the method efficiency. PMID- 2560133 TI - [Benign lesions of the salivary glands of the lip. Apropos of 3 cases]. AB - The paper reports data concerning three cases of benign salivary gland lesion of the lip in order to contribute to the number of case studies given the reduced frequency of this pathology. Patients underwent surgical exeresis of the lesions and no relapse was observed during follow-up. PMID- 2560134 TI - Isolation and characterization of herC, a mutation of Escherichia coli affecting maintenance of ColE1. AB - Two modes of ColE1 DNA replication are known, one dependent on RNase H, and the other RNase H independent. The cer114 mutant of the ColE1 replicon is defective in both modes and carries a single base pair alteration 95 bp upstream of the replication origin. An Escherichia coli mutant which restored maintenance of the cer114 replicon was isolated. This host suppressor mutant is defective in RNase H and carries a herC mutation located at 62 min of the E. coli chromosome. The herC mutation is recessive to its wild-type allele and supports maintenance of the mutant replicon in the absence of RNase H. The herC mutation alone conferred cold sensitive growth, suggesting that the herC gene product is essential for cell growth. The 1832 bp E. coli DNA fragment, containing the wild-type allele of the herC mutation, was cloned and an open reading frame for the HerC protein was determined. PMID- 2560135 TI - A genetic and molecular analysis of P-induced mutations at the glucose-6 phosphate dehydrogenase locus in Drosophila melanogaster. AB - We examined P factor induced mutations of the Zw gene of Drosophila melanogaster in order to learn more about the site specificity of such mutations. Approximately 70,000 chromosomes were screened using a powerful positive selection scheme. As only two mutants were discovered, Zw is a "cold spot" for transposable element insertion. One mutation involved a complex P element associated chromosomal rearrangement which was used to define the orientation of the gene with respect to the centromere of the X chromosome. The second mutation was either a simple, non-dysgenically induced point mutation or a very unstable insertion. PMID- 2560136 TI - Cloning and sequencing of the fbcF, B and C genes encoding the cytochrome b/c1 complex from Rhodopseudomonas viridis. AB - The complete nucleotide sequence of the genes encoding the Rieske FeS, the cytochrome b and the cytochrome c1 subunits of the ubiquinol-cytochrome c2 oxidoreductase from the photosynthetic purple bacterium Rhodopseudomonas viridis, and the derived amino acid sequences are presented. These three genes, fbcF, fbcB and fbcC, are located at contiguous sites of the genome. The DNA-deduced amino acid sequences are compared with known primary structures of corresponding proteins from other purple photosynthetic bacteria, as well as mitochondria, cyanobacteria and chloroplasts. PMID- 2560137 TI - Gene cloning shows the alpha-toxin of Clostridium perfringens to contain both sphingomyelinase and lecithinase activities. AB - The plc gene encoding the alpha-toxin (phospholipase C), an important virulence factor of Clostridium perfringens, has been cloned, sequenced and expressed in Escherichia coli. Transcriptional analysis of mRNAs produced in vivo by C. perfringens and E. coli, and in vitro using purified RNA polymerase from C. perfringens revealed that plc is transcribed constitutively from a single promoter situated about 100 nucleotides from the coding sequence. A T7 expression system was used to overproduce alpha-toxin in E. coli; enzymological studies with the amplified plc gene product unambiguously demonstrated that both lecithinase (phospholipase C) and sphingomyelinase activities were associated with this 43,000 dalton cytotoxin. The 370-residue alpha-toxin is haemolytic and shares sequence and functional homology with the two components of Bacillus cereus haemolysin, cereolysin AB, in which phospholipase C and sphingomyelinase activities are associated with different polypeptides. PMID- 2560138 TI - Lack of alpha-1-adrenergic receptor-mediated downregulation of angiotensin II receptors in neuronal cultures from spontaneously hypertensive rat brain. AB - Neuronal cells from Wistar Kyoto (WKY) and spontaneously hypertensive (SH) rat brains were established in culture to compare the expression of angiotensin II (Ang II) specific receptors and their regulation by norepinephrine (NE). Neurons from SH rat brains possess twice more Ang II specific receptors and expressed a proportional increase in Ang II stimulated [3H]-NE uptake compared with WKY neurons. NE caused a dose-dependent decrease in 125I-Ang II binding in WKY neurons, an effect not observed when neurons from SH rat brains were incubated with NE. These observations suggest that the lack of NE-induced downregulation of Ang II receptors in neuronal cultures is genetically regulated. PMID- 2560139 TI - Demonstration of an endogenous activator for the Na+, K(+)-ATPase system. AB - A heat-labile, non-dialysable and protease-sensitive endogenous activator (NaAF) capable of stimulating the Na+, K(+)-ATPase system has been demonstrated. The activator (NaAF) activity was partially enriched (about 10 fold) by dialysis (30 kDa cutoff) under negative pressure and pH 4.8 precipitation. The NaAF has been found to occur in the cytosolic fractions of tissues such as the kidney and brain from two different species (rabbit and pig) tested so far. Also, the factor from one tissue stimulates with equal efficacy the Na+, K(+)-ATPase systems of other tissues regardless of the species; thus demonstrating universal nature of the activator. Some degree of cross-reactivity was noted between the activating effects of this activator (for the Na+,K(+)-ATPase) and that for the H+,K(+) ATPase recently described (J. Biol. Chem. 262:5664-5670, 1987). The purified NaAF obtained from sephacryl S-300 column chromatography activates the pure renal medullary Na+,K(+)-ATPase in a dose dependent manner. PMID- 2560140 TI - Membrane transport and disease. AB - Four situations in which membrane transport is altered by disease are discussed: (a) non-specific leaks induced by poreforming agents; (b) glucose transport and cellular stress; (c) Ca2+-ATPase and hypertension; (d) Na+ channels and HSV infection. PMID- 2560142 TI - Automatic analysis of heart rate variation: II. Findings in patients attending an EMG laboratory. AB - An automatic method was used to measure the heart rate variation with breathing in patients with different neuromuscular conditions attending a laboratory of electromyography (EMG). The objective was to determine the frequency of abnormalities in various conditions and the relationship between R-R variation and different nerve conduction parameters. The percentages of reduced R-R variation were 73% in diabetics, 35% in Guillain-Barre syndrome, 22% in amyotrophic lateral sclerosis, and 50% in amyloidosis. R-R variation in diabetics was significantly correlated to most parameters of nerve conduction. In Guillain Barre syndrome it correlated significantly with the ulnar and median M-response amplitudes. Most patients with myopathies showed normal R-R variation. PMID- 2560141 TI - Patterns of T cell receptor gamma gene rearrangements in human CD3+ clones derived from WT31- or Leu7+ cells in relation to non-MHC-restricted cytotoxic activity. AB - Clones were obtained from human peripheral blood WT31-, WT31-CD4-8-, CD4-8- or Leu 7+ cells in the presence of interleukin 2 and phytohaemagglutinin. Almost all clones were CD3+, about 50% were CD4-8- and all clones tested derived from WT31- remained WT31-, indicating that they were expressing a gamma/delta heterodimer in association with CD3. Some clones derived from CD4-8- cells expressing CD3 were WT31- and some were WT31+. All CD3+ clones had T cell receptor (TCR) gamma gene rearrangements; most also had their TCR beta genes rearranged, including all clones derived from Leu 7+ cells. TCR gamma gene rearrangements were noted involving all five known J segments. There was a tendency for V gene segments from the VII and VIII subgroups to be rearranged to J gamma 2 less often than those from the more 5' VI subgroup. Two clones definitely had one rearrangement to C gamma 1 and one to C gamma 2. When clones derived from WT31- cells were considered, the only obvious relationship which emerged was that all clones with both chromosomes rearranged to C gamma 2 had low or negligible cytotoxic activity against natural killer (NK)-sensitive and NK-resistant targets. Several of these clones were expressing CD8 on about 30% of cells. Most clones with rearrangements involving only C gamma 1 had high non-MHC-restricted cytotoxicity while those with at least one C gamma 1 rearrangement had either high or low activity. The only exceptions noted were a clone with a single V9JP rearrangement and a clone with a V9JP and a VI/IIIJP1 rearrangement, which both had low activity. A similar pattern was also found with most clones derived from Leu 7+ cells. The data are consistent with the participation of most types of disulphide-linked (C gamma 1) gamma/delta heterodimers in non-MHC-restricted cytotoxic activity mediated by CD3+ gamma/delta + T cell clones. PMID- 2560143 TI - Functional evaluation of acute vincristine toxicity in rat skeletal muscle. AB - Vincristine sulfate (VCR) was administered intravenously to rats at doses of 0.25, 0.5, and 0.75 mg/kg. During the first week following VCR treatment, extensor digitorum longus (EDL) muscle contraction strength and fiber electrophysiologic parameters were measured. At all doses tested, VCR strongly reduced twitch and tetanic tension. EDL fiber resting membrane potential was affected in a dose-dependent manner, and membrane depolarization was associated with the loss of excitability. Local membrane hyperpolarization by intracellular current application restored the capacity to produce action potential (AP). However, to elicit APs with a normal rate of rise, polarizing current had to be maintained for 3-5 minutes, indicating that the removal of Na+ channel inactivation followed a slow kinetics. Minor alterations in spontaneous synaptic transmission and in evoked transmission during high-frequency repetitive stimulation were seen only at the highest dose. It is suggested that VCR impairs skeletal muscle function by affecting primarily the contractile apparatus, whereas sarcolemmal alterations are evident at increased doses of the drug. PMID- 2560144 TI - Microsomal reduction of alloxan produces alloxan anion radicals. PMID- 2560145 TI - [The HA-Ti system]. AB - Some concepts and techniques are discussed while stressing the collaboration between specialists to design a new type of implant. Important aspects are: a simple implantation procedure, a wide field for indication and non-traumatic surgical placement. PMID- 2560146 TI - Characterization of the calcitonin gene-related peptide receptor in mouse T lymphocytes. AB - We have identified and characterized specific binding for calcitonin gene-related peptide (CGRP) in mouse T lymphocytes. The binding of 125I-CGRP to mouse lymphocytes was reversible and the rate of dissociation of 125I-CGRP increased with the addition of the guanine triphosphate nucleotide analog, Gpp(NH)p. Saturation experiments, and Scatchard analysis indicated two classes of binding sites for CGRP; the apparent dissociation constants (KD) were 3.5 X 10(-10)M for high affinity binding sites (Bmax, 265 sites/cell) and 4.8 X 10(-8)M for low affinity binding sites (Bmax, 13,000 sites/cell). The KD value for the high affinity binding sites is roughly comparable to the IC50 and ED50 values for inhibition of T lymphocyte proliferation and increase in the cyclic AMP concentration in these cells, respectively. 125I-CGRP bound to mouse T lymphocytes was displaced by unlabeled CGRP with an IC50 value of 3.2 X 10(-10)M; salmon or human calcitonins did not inhibit the specific binding up to 1 X 10( 6)m. Our studies suggest that CGRP may be an important immunoregulatory molecule, and implicate it in the regulation of T cell function. PMID- 2560147 TI - Histochemical changes in enzymes of energy metabolism in the dentate gyrus accompany deafferentation and synaptic reorganization. AB - The dentate gyrus of adult rats was examined histochemically for cytochrome oxidase and lactate dehydrogenase activity after unilateral lesions of the entorhinal cortex. In normal animals, synaptic terminal fields of the perforant pathway from the entorhinal cortex show high levels of cytochrome oxidase activity (the other two-thirds dentate molecular layer), whereas terminal zones of the commissural and associational fibers show high levels of lactate dehydrogenase activity (the inner one-third dentate molecular layer). Lesions of the entorhinal cortex result in a significant reduction in staining for cytochrome oxidase in the deafferented outer molecular layer of the dentate gyrus. The changes become prominent at 16-24 h after the lesion and persist until 90 days, the longest post-lesion survival time studied. In the non-deafferented inner zones ipsilateral to the lesion, there is an increase in staining for cytochrome oxidase and lactate dehydrogenase at 24 h post-lesion that disappears by days 2-4. From 8 to 90 days post-lesion, the band of high reactivity for lactate dehydrogenase in the inner molecular layer spreads approximately 40 microns into the overlying deafferented zone. This expansion parallels the expansion of the commissural and associational terminal fields into the adjacent deafferented molecular layer. Thus, lesion-induced synaptogenesis in the dentate gyrus is accompanied by a corresponding change in enzyme activity. The results indicate that the pattern of activity of enzymes involved in energy metabolism in the dentate gyrus depends on the distribution of pathway-specific synaptic input. PMID- 2560148 TI - Modulatory effects of serotonin on excitatory amino acid responses and sensory synaptic transmission in the ventrobasal thalamus. AB - Excitatory amino acid receptors are thought to mediate sensory input to the ventrobasal thalamus. There is evidence for a brainstem serotonergic projection to the ventrobasal thalamus which may have a modulatory role. The possibility that serotonin may selectively modulate responses to excitatory amino acid receptor agonists, and its effects on sensory synaptic transmission has been examined in the rat ventrobasal thalamus in vivo. Iontophoretic ejection of serotonin at low currents produced a marked facilitation of responses to excitatory amino acids. In contrast, excitatory responses to cholinomimetic agonists were attenuated. Synaptic transmission was concomitantly enhanced or unchanged in these circumstances. Higher serotonin ejection currents reversed the facilitation, or inhibited excitatory amino acid responses and synaptic transmission. It is concluded that serotonin can modulate responses to excitatory amino acids relatively selectively and that synaptic transmission of somatosensory information through the ventrobasal thalamus may be susceptible to brainstem serotonergic modulation. PMID- 2560149 TI - Quantitative histochemical analysis of cytochrome oxidase in rat dorsal root ganglia and its co-localization with carbonic anhydrase. AB - A quantitative histochemical method was developed and standardized and then used to characterize the heterogeneity of cytochrome oxidase activity among primary afferent neuronal cell bodies in dorsal root ganglia of rat. In addition, the relationship between cytochrome oxidase and carbonic anhydrase activities in these neurons was determine. In tests of the procedure, the density of cytochrome oxidase reaction product evaluated repeatedly in individual neurons within sections of ganglia was found to increase linearly over incubation periods of up to 6 h. The heterogeneity in cytochrome oxidase activity in ganglia was not simply a reflection of the heterogeneity in ganglion cell sizes. On the whole, each class of ganglion cell exhibited the full range of staining densities encountered but intense staining was observed in many more large type A cells than small type B cells. The latter, together with their termination fields within the substantia gelatinosa of the spinal cord, were lightly stained. A significant positive correlation was found between neuronal size and staining density (r = 0.43). However, the large scatter in the plot of these two variables suggests that the expression of cytochrome oxidase in sensory neurons is governed to a considerable extent by properties of these neurons that are unrelated to their size. Analysis of cytochrome oxidase and carbonic anhydrase activities in the same ganglion cells revealed that all neurons with dense staining for the oxidase were anhydrase positive. Conversely, however, some intensely anhydrase positive cells exhibited only light staining for cytochrome oxidase. The heterogeneity of cytochrome oxidase activity among neurons in dorsal root ganglia may be related to the steady state electrophysiological activity of distinct populations of sensory neurons which in turn may be related to the specific sensory modalities these populations transmit. The observation that some neurons with the greatest abundance of carbonic anhydrase do not contain high or even moderate levels of cytochrome oxidase suggests some degree of dissociation between the functional requirement for carbonic anhydrase in sensory neurons and the rate of energy expenditure in these cells. PMID- 2560150 TI - Analysis of parvalbumin and calbindin D28k-immunoreactive neurons in dorsal root ganglia of rat in relation to their cytochrome oxidase and carbonic anhydrase content. AB - Histochemical and immunohistochemical techniques were used to determine relationships between the parvalbumin or calbindin D28k content and the cytochrome oxidase or carbonic anhydrase activity of neurons in lumbar dorsal root ganglia in rat. Subpopulations of dorsal root ganglion neurons that displayed parvalbumin- or calbindin D28k-immunoreactivity were classified as containing either light, moderate or dense histochemical reaction product for cytochrome oxidase and either a positive or negative reaction for carbonic anhydrase. It was found that approximately 90% of all parvalbumin and calbindin D28k-immunoreactive cells exhibited dense staining for cytochrome oxidase and that 87% of parvalbumin- and 76% of calbindin D28k-immunoreactive cells were positive for carbonic anhydrase. Conversely, 85% of all cells with a dense cytochrome oxidase reaction contained parvalbumin and calbindin D28k. Although not quantified, it appeared that many, but not all, carbonic anhydrase-positive cells contained parvalbumin or calbindin D28k. These results indicate the existence of a subpopulation of primary sensory neurons that contains parvalbumin and calbindin D28k and that expresses high levels of cytochrome oxidase and carbonic anhydrase activity. It is suggested that primary afferent neurons with this cytochemical profile transmit a sensory modality that requires them to discharge rapidly and/or frequently. The existence of a subpopulation of carbonic anhydrase-positive cells that lack immunoreactivity for parvalbumin or calbindin D28k suggests that the role of carbonic anhydrase in some sensory neurons is unrelated to functions requiring these calcium binding proteins. PMID- 2560151 TI - [Bronchial carcinoma in subjects under 40]. AB - We investigated a series of 30 patients younger than 40 years with histologically proven bronchogenic carcinoma. Most patients were symptomatic for a mean duration of 3 months before examination. The most common cell types were squamous carcinoma in 10 patients, oat cell carcinoma in 9 patients, and adenocarcinoma in 5 patients. The disease was categorized as Stage I in 3 patients, Stage II in 3, Stage III in 17, and Stage IV in 7. Ten patients underwent resection, whereas 3 patients were inoperable at surgery. The average length of survival for the nonoperated patients was 4.2 months (range 1 to 16 months). The mean survival for the surgically treated patients was 23.3 months (range 0 to 84 months). Extent of resection did not have any statistical significance in the survival rates which were mainly affected by staging of disease. PMID- 2560152 TI - [Cytomegalovirus transmission following a blood transfusion]. AB - In order to assess the incidence of anti-CMV antibodies in blood donors and the possible transmission of infection in blood transfusions, 900 donors, 60 patients serum negative or positive patients given serum positive blood and 5 immature serum negative patients given serum negative blood were examined. At 67.11% the incidence of anti-CMV antibodies in the donors was in line with reports in the literature. It was also noted that reinfection arose when serum positive blood was given (36.36% of the serum negative patients and 10.52% of the serum positive ones developed anti-CMV antibodies indicating infection or reinfection; none of the 5 immature patients revealed anti-CMV antibodies). It is concluded that the routine search for anti-CMV antibodies is impracticable given the high incidence of serum-positive donors and that the technique should be reserved for particular cases: the young, the pregnant, transplant patients, immunodepressed subjects. However the best way to prevent the transmission of disease remains the avoidance of unnecessary transfusions. PMID- 2560153 TI - [Hepatocellular carcinoma (HCC): the long-term response to tamoxifen. A clinical case report and review of the literature]. AB - A 52 years old female with hepatocellular carcinoma (HCC) was treated successfully with Tamoxifen. The tumor involved IV hepatic segment with hilar extension and biliary obstruction, was unresectable, and had been pretreated with hormone-chemotherapy. Tamoxifen treatment induced a PR of 6 months, with normalization of serum bilirubin, reduction of alfa-fetoprotein level and improvement of PS, and was free of toxicity. At disease progression intra arterial chemotherapy with Cis platinum (CDDP) and 5-FU gave a further 4 months PR, until disease progression and exitus in hepatic coma. Tamoxifen therapy, even in the absence of E.R. assay is a useful tool in the management of HCC patients. Further randomized studies are necessary to ascertain the role of Tamoxifen in the treatment of HCC. PMID- 2560154 TI - 99Tcm(v)-DMSA planar scintigraphy: does it have a role in the management of patients with head and neck squamous carcinoma? AB - 99Tcm(v)-DMSA is a new tumour-imaging agent which has recently been proposed as a scintigraphic marker for head and neck squamous cell carcinoma (SCC). Seventy seven patients were studied prospectively, of whom 58 had a history and diagnosis of head and neck SCC. All patients were examined, imaged using 99Tcm(v)-DMSA planar scintigraphy and then followed up clinically. In addition, 35 patients were followed up with scintigraphy (81 studies). Scintigraphy was less sensitive and less accurate than clinical examination for the overall detection of patients with SCC, for the detection of patients with SCC at presentation and for the detection of patients with primary tumours, possible nodal disease and with residual and recurrent disease following surgery and irradiation. Approximately 50% of patients exhibited positive uptake of 99Tcm(v)-DMSA in the salivary glands following radiotherapy. Although 99Tcm(v)-DMSA is accumulated at sites of head and neck SCC, its inability to detect low volume disease and apparent low specificity following surgery and irradiation means it has no role to play in the routine evaluation of patients with head and neck SCC. PMID- 2560155 TI - Regional lung microvascular permeability using dual isotope scintigraphy. AB - We have extended the dual isotope probe technique of Basran et al. for the measurement of pulmonary microvascular permeability (PMVP) to include gamma camera data acquisition and functional imaging of the plasma protein accumulation index (PPA) throughout the lung fields. The study group consisted of 11 patients with possible increased PMVP following the drainage of a pleural effusion or the evacuation of air from a pneumothorax, and 11 control patients. The PPA was calculated (1) for probe data, (2) on a pixel by pixel basis for the camera data, the results being stored in a functional image and (3) for four pixel x four pixel regions of interest positioned over the lung fields using the functional image and raw data as a guide. Functional images of the control group showed uniformly low PPAs throughout the lung fields. Nine of the eleven patients in the effusion/pneumothorax group showed discrete areas of increased PPA on the functional images. In the effusion/pneumothorax group, the re-expanded lung four pixel x four pixel ROI PPA values were significantly higher than the nonexpanded lung ROI values (p less than 0.001). The re-expanded lung ROI values were also significantly higher than the mean of the left and right lung ROIs in the control group (p less than 0.01). Five of the eleven patients in the effusion/pneumothorax group had probe PPAs that differed significantly from zero. We believe that probe positioning problems in the absence of sufficient clinical guidelines were largely responsible for there being no overall significant difference in the probe PPAs between the two study groups. PMID- 2560156 TI - Bilateral optic nerve compression as a mechanism for the Foster Kennedy syndrome. AB - An 11-year-old boy with a juvenile nasopharyngeal angiofibroma developed optic disc pallor in one eye and optic disc edema in the other eye (Foster Kennedy Syndrome [FKS]). The mechanism was believed to be bilateral, asymmetric optic nerve compression. Review of the 36 previously reported cases of FKS revealed that 12 cases (33%) were probably also caused by bilateral optic nerve compression. Only eight (22%) of the cases satisfied Foster Kennedy's original hypothesis for the pathogenesis of his syndrome, namely, direct compression of one optic nerve causing atrophy and increased intracranial pressure causing contralateral papilledema. In 15 (41%) reported cases of FKS, descriptions were inadequate to determine a mechanism, while two (5%) were probably caused by long standing increased intracranial pressure without direct optic nerve compression. The authors believe that as sophisticated imaging permits earlier diagnosis and more precise localization, most future cases of FKS caused by mass lesions will be found to result from bilateral direct optic nerve compression. PMID- 2560157 TI - [2 cases of glomangioma of rare location]. AB - The nonmalignant vascular tumor named glomangioma of rare localization in larynx and tympanic cavity were described. The treatment consisted of surgery. PMID- 2560158 TI - [Zona in a 6-month-old infant. Apropos of one case]. AB - The authors report an unusual case of zona in a 6-month-old infant whose mother was affected by varicella on the fourth month of pregnancy. The course and outcome were benign. PMID- 2560159 TI - [Present status of vaccines in 1989]. AB - The authors describe 2 new vaccines now available in France: one is the GenHevac, an hepatitis B vaccine, the first virus recombinant vaccine; the other one is the Typhim Vi, a polysaccharide typhoid vaccine. Three other vaccines are currently used in foreign countries and will be soon available: the Hemophilus influenzae vaccine, the acellular pertussis vaccine and the varicella vaccine. Rotavirus and Cytomegalovirus vaccines are studied for their clinical efficacy. PMID- 2560160 TI - Regulation of Ca2+ current in frog ventricular myocytes by the holding potential, c-AMP and frequency. AB - The whole-cell patch-clamp technique was used to study the effects of holding potential and frequency on the Ca2+ current in frog ventricular myocytes. INa was blocked by TTX, and ica was activated with depolarizing clamps from different holding potentials. Variation of the holding potential revealed three new effects on ica: (1) At -40 mV iCa declined with a time constant of 15 min, while at -90 mV, this irreversible decline (run down) in iCa did not occur. (2) The decline of iCa at -40 mV was biphasic: run down was preceeded by a slow inactivation with a time constant of 40 s, which was reversible upon returning the holding potential to -90 mV. (3) Increasing the frequency of the clamp pulses from 0.1 to 1 Hz led to a rapid decline of iCa when the holding potential was positive to -60 mV, but at -90 mV had either no effect or increased iCa by 35%, if c-AMP was included in the dialyzing solution. On the other hand, c-AMP did not alter the time course of the run down and the slow inactivation. Replacement of extracellular Ca2+ by Ba2+ markedly slowed iCa kinetics, but did not change the very slow inactivation or the frequency-induced enhancement of iCa. Injection of c-AMP led to a transient increase of iCa. The phosphodiesterase inhibitor theophylline enhanced the amplitude of the transient and slowed its decay. This effect was mimicked by increased frequency. It is concluded that frequency-induced enhancement of iCa is highly dependent on the holding potential, independent of Ca2+, and may involve elevation of the intracellular level of c-AMP via inhibition of phosphodiesterase activity. The new type of very slow inactivation is probably under direct voltage control and independent of Ca2+ and c-AMP. PMID- 2560161 TI - Chloride current activated by cyclic AMP and parathyroid hormone in rat osteoblasts. AB - In primary cultures of rat osteoblasts, studied with the whole-cell configuration of the patch-clamp technique, 8-bromo-cyclic AMP (8BrcAMP) forskolin (FS) and 1 34 parathyroid hormone (PTH) were shown to activate a Cl conductance. This conductance shows a pronounced outward rectification, even with symmetrical Cl concentrations. It is blocked partially and reversibly by 4,4' diisothiocyanatostilbene 2,2'-disulfonic acid (DIDS) or diphenylcarboxylate (DPC). The blockade induced by DIDS is time- and voltage-dependent. The Cl responses to FS and PTH develop slowly, after a delay of several seconds and are very slowly reversible. These responses were observed only in a fraction of the cells tested and their detection was favoured by cell dialysis. This Cl current should be taken into account for studying possible modulations of the voltage gated Ca currents of osteoblasts. It is suggested that its physiological role may be related to the well-known morphological changes induced by PTH in osteoblasts. The cyclic AMP-sensitivity, the outward rectification and the sensitivity to dialysis of this Cl current are reminiscent of the properties of the cystic fibrosis-sensitive Cl channels of epithelial cells. PMID- 2560162 TI - Ischemic poison lysophosphatidylcholine modifies heart sodium channels gating inducing long-lasting bursts of openings. AB - The effects of lysophosphatidylcholine (LPC) on Na channels in inside-out patches of adult rat ventricular cells using the patch-clamp technique have been investigated. Application of LPC (9-25 microM) from the inner side of membrane for 4-15 min caused a reduction of averaged Na current (INa) peak and prolonged the time course of inactivation in the potential range of -50 to -10 mV. Analysis of single channel behaviour revealed that after 30-50 min of exposure, in addition to normally functioning Na channels with short openings, LPC induced long-lasting bursts of Na channel openings (up to the 300 ms duration of the test pulses). This resulted in an appearance of noninactivated component of INa. The slope conductance of these modified channels remained the same as in control (11.3 pS - control; 11.6 pS - LPC-treated). The dwell time for modified channels increased significantly. PMID- 2560163 TI - Activation of K+ currents in cultured Schwann cells is controlled by extracellular pH. AB - We analyzed the pH dependence of K+ currents recorded with the patch-clamp technique from cultured Schwann cells obtained from mouse dorsal root ganglia. Currents were activated at potentials more positive than -50 mV which was close to the resting membrane potential. Current amplitudes were affected by a change in extracellular pH (pHo), being increased at alkaline, and decreased at acidic pHo. The strongest effect of a pHo change was observed on currents activated close to the resting membrane potential suggesting a functional role for the pH sensitivity of K+ currents. Analysis of the time course of current activation at different pHo values led to the conclusion that the pH-sensitivity of K+ currents in Schwann cells is due to changes in surface charges shifting the potential sensed by the gating process of the channel. The reversal potential of the currents was not affected by a change in pHo. This observation and the finding that even a strong acidification to a pHo value of 5.0 did not lead to a blockade of the fully activated channel, indicate that the pH-sensitive charges are not located in the channel pore. Under the assumption that pHo changes in a peripheral nerve are associated with nerve activity as in the optic nerve, the pH sensitive K+ channel in Schwann cells could serve to facilitate the spatial buffering of extracellular K+. PMID- 2560164 TI - Secretagogue and second messenger-activated Cl- permeabilities in isolated pancreatic zymogen granules. AB - Previous studies of enzyme secretion from isolated pancreatic acinar cells and of isolated zymogen granules (ZG) have reported that both a Cl- and a K+ permeability are present on the ZG membrane. It has been suggested that ion influx via these permeability pathways, followed by water movement is required for granular swelling which appears to be intimately related to exocytosis. However, little is known about the regulation of these pathways by secretagogues. Evidence suggests that cAMP-protein kinase A and diacylglycerol-protein kinase C are second messengers in stimulation of exocytosis. In the present study we have examined ion permeability pathways in ZG isolated from control cells and from cells pretreated with the acetylcholine analog carbachol (Cch), with the peptide hormone cholecystokinin (CCK) and with second messengers of hormone action such as cAMP and the diacylglycerol analog 12-O-tetradecanoyl phorbol-13-acetate (TPA). Ion and water influx rates in ZG and consequent swelling and lysis of granules was monitored by measuring changes in optical densities of ZG suspensions at 540 nm following additions of the electrogenic or electroneutral ionophores valinomycin and nigericin, respectively. The data show that both a Cl- conductance and an anion exchange pathway are present in the granule membrane. Both pathways are activated by pretreatment of isolated cells with CCK or of isolated permeabilised cells with cAMP, whereas only the Cl- conductance is increased by pretreatment with Cch or with TPA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2560165 TI - Effect of isoprenaline on active Na transport in sheep cardiac Purkinje fibres. AB - The effect of isoprenaline (ISO; 5.10(-8) to 10(-6) M) on active Na transport was studied in depolarized sheep cardiac Purkinje fibres. Membrane current (I) and intracellular Na activity were measured simultaneously during enhanced Na pumping in voltage clamped preparations. ISO stimulated enhanced active Na transport but did not affect membrane current or intracellular Na concentration (ciNa) in the steady state under the chosen experimental conditions. The stimulatory effect of ISO was mediated by beta 1-adrenoceptors via a cAMP dependent pathway. The effect depended on the extracellular K concentration (coK) and was inhibited by external Ba ions. Complementary experiments on isolated sheep Purkinje cells revealed no ISO induced alteration of the Na pump current. The mechanism of the ISO induced stimulation of enhanced Na pumping in sheep Purkinje fibres probably involves an augmented K efflux. A direct effect on the pump molecule seems unlikely. PMID- 2560167 TI - Direct modulation of voltage-dependent calcium channels by muscarinic activation of a pertussis toxin-sensitive G-protein in hippocampal neurons. AB - Acetylcholine (Ach) reversibly reduces the high voltage-activated (HVA) calcium (Ca) current in hippocampal neurons. Pretreatment of the cells with pertussis toxin (PTX) abolishes the Ach effect, suggesting that PTX-sensitive GTP-binding regulatory proteins (G-proteins) are involved in the signal transduction mechanism that links Ach receptor activation to inhibition of Ca channel activity. This effect is mimicked by intracellular application of the nonhydrolyzable GTP analog GTP gamma S. Intracellular application of purified G proteins restored the response to Ach in PTX-treated cells. Furthermore, Ach inhibits the Ca current independently of the presence of cyclic AMP and of the protein kinase C inhibitor H-7 and neither does the Ach effect on the Ca current seem to be correlated to a transient increase in intracellular Ca. Our results suggest that activation of the alpha-subunit of the PTX-sensitive G-protein could directly modulate the HVA Ca channel without involving second messenger systems. PMID- 2560166 TI - An electrophysiological investigation of the characteristics and function of GABAA receptors on bovine adrenomedullary chromaffin cells. AB - The characteristics and function of gamma-aminobutyric acidA (GABAA) receptors expressed on bovine chromaffin cells in culture have been investigated using patch-clamp techniques. In voltage-clamped whole-cells, locally applied GABA (100 microM) evoked a transmembrane chloride current which demonstrated outward rectification. The amplitude of such currents was reversibly suppressed by the GABAA receptor antagonists bicuculline, picrotoxin and RU5135, and enhanced by the general anaesthetic propanidid. Glycine (100 microM) and baclofen (100 microM) were ineffective as agonists. In support of a physiological role for GABA in the adrenal medulla, the co-existence of GABAA and nicotinic acetylcholine (ACh) receptors was demonstrated on whole cells and outside-out membrane patches. Ionophoretically applied GABA reduced the amplitude of depolarization and action potential discharge occurring in response to locally applied ACh (100 microM), but had no effect upon the underlying ACh-induced current. In addition, an excitatory action of GABA was demonstrated by recording action potential waveforms in cell-attached patches. The results are discussed in the context of a GABA-ergic regulation of catecholamine secretion. PMID- 2560169 TI - Inhibition of voltage-dependent Na+ and K+ currents by forskolin in nodes of Ranvier. AB - The effect of forskolin on voltage-activated Na+ and K+ currents in nodes of Ranvier from the toad, Bufo marinus, has been examined using the vaseline-gap voltage-clamp technique. Peak Na+ currents (INa) were reduced by 35% and the rate of decline of Na+ current during continuous depolarization was accelerated following treatment with 450 microM forskolin. However, the voltage-dependence of steady-state inactivation as well as the rate of recovery from fast inactivation remained unchanged. Upon repetitive depolarization at 1-10 Hz, a further inhibition of INa (approximately 60%) was observed. This use-dependent or phasic inhibition recovers slowly at -80 mV (tau approximately 13 s) and had a voltage dependence like that of activation of the Na conductance. Near maximal steady state phasic inhibition occurred with depolarizing pulse durations of only 4 ms, consistent with a direct involvement of the open Na+ channel in the blocking process. Inhibition of the delayed K+ current (IK) was characterized by a concentration-dependent reduction in steady-state current amplitude (IC50 approximately 80 microM) and a concentration-independent acceleration of current inactivation. A similar inhibition of IK was obtained with 1,9-dideoxyforskolin, a homolog which does not activate adenylate cyclase (AC). The results suggest that the inhibition IK and perhaps INa follows directly from drug binding and is not a consequence of AC activation. PMID- 2560168 TI - Beta-adrenergic modulation of transient inward current in guinea-pig cardiac myocytes. Evidence for regulation of Ca2(+)-release from sarcoplasmic reticulum by a cyclic AMP dependent mechanism. AB - Transient inward current (Iti) indicating Ca2(+)-release from the sarcoplasmic reticulum and L-type Ca2(+)-current (ICa) were studied in atrial and ventricular myocytes from hearts of adult guinea-pigs by means of whole-cell voltage-clamp. The increase of ICa caused by beta-adrenergic stimulation using isoprenaline (ISO) or related experimental manoeuvres such as superfusion with forskolin (FORSK) was used as a qualitative monitor of an increase of intracellular cAMP. Changes of Iti were used to manifest changes of sarcoplasmic Ca2(+)-release. In myocytes dialysed with citrate-based (60 mM) pipette filling solution containing 100 microM EGTA spontaneous transient inward currents were recorded at a constant holding potential of -50 mV in the majority of myocytes. Superfusion with a solution containing ISO (greater than or equal to 5 x 10(-8) M) increased the amplitude of spontaneous Iti and reduced its time-to-peak. The effects of ISO on Iti developed in parallel to stimulation of ICa. In myocytes which did not show spontaneous cyclic Ca2(+)-release in the above condition, this could be evoked de novo by ISO. Spontaneous Iti was suppressed in the majority of cells by increasing the concentration of EGTA in the dialysing solution to 200 microM. Brief (50 ms) activation of ICa by voltage steps from -50 to +10 mV usually failed to trigger Ca2(+)-release from the SR. The increase of ICa-amplitude upon administration of ISO went ahead with the induction of Ca2(+)-release by brief activation of ICa. The effects of ISO could be mimicked by FORSK or intracellular dialysis with 3'5'-cyclic adenosine monophosphate. The effects on ICa and SR Ca2(+)-release were dependent o the concentration of the stimulating substance. In a given cell changing superfusion from a low to a high concentration of ISO or FORSK resulted in an increase of the number of Ca2(+)-release events per number of Ca2(+)-currents elicited and a shortening of time-to-peak of Iti's. The stimulating effects of ISO or FORSK on Ca2(+)-release were only partially due to an increase of the triggering ICa. Ca2(+)-currents too small to trigger Ca2(+) release before beta-adrenergic stimulation could evoke Ca2(+)-release after augmentation of intracellular cAMP. Whereas the effects of ISO and FORSK on ICa were reversible, the stimulatory effects on Ca2(+)-release persisted after washing out the substances. The results give support to the hypothesis that beta adrenoceptor-mediated positive inotropic and arrhythmogenic effects are, at least partly, due to a cyclic AMP-dependent regulatory mechanism modulating sarcoplasmic Ca2(+)-release. PMID- 2560170 TI - Changes in Na channel properties of frog and rat skeletal muscles induced by the AaH II toxin from the scorpion Androctonus australis. AB - The effects of the mammal toxin II isolated from the venom of the scorpion Androctonus australis Hector (AaH II) were studied under current and voltage clamp conditions in frog (semitendinosus) and rat (fast e.d.l. and slow soleus) skeletal twitch muscle fibres. In both species, AaH II induced a dose-dependent prolongation of the action potential (AP) leading at saturating concentration to APs with long plateaus of about 1.5 s in frog and 5 s in rat e.d.l. and soleus fibres. The concentrations to induce 50% of the maximal effect (K0.5) were 9.1 x 10(-9) M in the frog and 1.4 x 10(-9) M in the rat. AaH II increased the time constants of inactivation of the peak Na current and induced a maintained Na current that was greater in rat e.d.l. and soleus (31.6% of peak current amplitude at -30 mV; K0.5 = 0.8 x 10(-9) M) than in frog (16.5%; K0.5 = 15.5 x 10(-9) M) muscles. Peak and maintained Na currents were TTX-sensitive and had identical threshold and reversal potentials. The half-maximum maintained permeability occurred at a potential 20 mV more positive than the peak permeability. Recovery from inactivation and steady-state inactivation of the inactivating Na current remained unchanged. The maintained current deactivated with normal fast kinetics. The action of the toxin reversed poorly on washout but could be largely removed by conditioning depolarizations more positive than the reversal potential of the Na current. Our results suggest that, in vertebrate skeletal muscle fibres, AaH II affects all the Na channels and are consistent with the hypothesis that the maintained current originates from a reopening of previously inactivated Na channels. PMID- 2560173 TI - Effect of posttraining and pretest beta-endorphin and ACTH administration in normal and protein malnourished rats. AB - Rats were submitted to a normal (25% casein) or a low protein diet (8% casein) from the day of birth until the age of 110 to 120 days. Hypothalamic beta endorphin-like immunoreactivity was lower in the animals raised and maintained with the low protein diet, and, in addition, it did not respond to training in a step-down inhibitory avoidance task with or without footshock with a depletion, as was the case with the normal diet animals. In the animals submitted to the normal protein diet posttraining ACTH (0.2 micrograms/kg) and beta-endorphin (1.0 micrograms/kg) caused retrograde amnesia of a step-down inhibitory avoidance task, and pretest administration of these substances had no effect of its own, but was able to reverse the amnesia induced by their previous posttraining administration. In the animals submitted to the low protein diet, results were similar except that pretest beta-endorphin caused amnesia on its own. On the basis of previous findings which suggest that pretest actions of ACTH and beta endorphin depend on their endogenous release at the time of training, the present results are compatible with a malfunction of the brain beta-endorphin system in the undernourished animals. PMID- 2560171 TI - The in vivo and in vitro effect of calmodulin antagonists on the renal actions of 25(OH) vitamin D3 in the rat. AB - Previous work from this laboratory has demonstrated that 25(OH) vitamin D3 [25(OH)D3] acutely suppresses the phosphaturic action of parathyroid hormone (PTH) and interferes with the PTH-induced activation of adenylate cyclase (AC). Calmodulin inhibitors block vitamin D-induced Ca2+ transport in the gut and phosphorus uptake in renal BBMV's. We have examined whether calmodulin antagonists affect the renal action of 25(OH)D3. Acute clearance experiments were performed in PTH-infused parathyroidectomized rats receiving 25(OH)D3 after pretreatment with trifluoperazine (TFP) or promethazine (P). In vitro PTH-induced activation of renal AC was also studied in membrane preparations from pretreated rats in the presence of 25(OH)D3. 25(OH)D3 reduced the PTH-stimulated increase in fractional excretion of phosphorus (CP/CIn) from 0.292 +/- 0.024 to 0.195 +/- 0.018 (p less than 0.005) and urinary cAMP from 149.3 +/- 20.3 to 78.1 +/- 10.4 pmol/min (p less than 0.01) and also blunted AC activation in vitro. TFP but not P abolished the effects of 25(OH)D3 both in vivo and in vitro. R 24571 also abolished the in vitro effect of 25(OH)D3. Thus, (1) TFP abolishes both the antiphosphaturic and the AC/cAMP-related actions of 25(OH)D3, (2) P does not have these effects, and (3) R 24571 abolishes the in vitro effect of 25(OH)D3. These results suggest that the antiphosphaturic effect of 25(OH)D3 acting via the AC/cAMP system may be calmodulin dependent. PMID- 2560172 TI - Li+ inhibition of membrane current responses to epinephrine in guinea-pig ventricular cells. AB - Membrane currents of guinea-pig ventricular myocytes were recorded using the whole-cell voltage clamp method. The epinephrine-induced increase in Ca2+ current (2.9 +/- 0.5 times control) was reduced (1.8 +/- 0.3 times) by replacing Na+ with Li+ in the bathing solution. In addition, 0.5 microM epinephrine increased a time independent membrane conductance in the Na+ external solution, having a reversal potential of -19 +/- 3 mV (epinephrine-induced current). In the Li+ external solution, however, 0.5 microM epinephrine failed to induce the epinephrine induced current. The findings are consistent with the reported Li+ inhibition of GTP-binding protein and/or adenylate cyclase. PMID- 2560174 TI - Effects of bradykinin on PC-12 cell differentiation. AB - PC-12 cells are used as a model for neuronal differentiation because they assume a neuronal phenotype, including the extension of neurites, when exposed to nerve growth factor (NGF). The present results show that bradykinin (BK) also causes PC 12 cells to extend neurites. In addition, BK potentiates the neurite-extending effect of nerve growth factor (NGF), an action which is attenuated by a BK antagonist. The potentiation of neurite extension produced by the combination of BK and NGF may be mediated at the receptor level, as indicated by an NGF-induced alteration of BK binding. PMID- 2560175 TI - Biological activities of des-His1[Glu9]glucagon amide, a glucagon antagonist. AB - Hyperglycemia in diabetes mellitus is generally associated with elevated levels of glucagon in the blood. A glucagon analog, des-His1[Glu9]glucagon amide, has been designed and synthesized and found to be an antagonist of glucagon in several systems. It has been a useful tool for investigating the mechanisms of glucagon action and for providing evidence that glucagon is a contributing factor in the pathogenesis of diabetes. The in vitro and in vivo activities of the antagonist are reported here. The analog bound 40% as well as glucagon to liver membranes, but did not stimulate the release of cyclic AMP even at 10(6) higher concentration. However, it did activate a second pathway, with the release of inositol phosphates. In addition, the analog enhanced the glucose-stimulated release of insulin from pancreatic islet cells. Of particular importance were the findings that the antagonist also showed only very low activity (less than 0.2%) in the in vivo glycogenolysis assay, and that at a ratio of 100:1 the analog almost completely blocked the hyperglycemic effects of added glucagon in normal rabbits. In addition, it reduced the hyperglycemia produced by endogenous glucagon in streptozotocin diabetic rats. Thus, we have an analog that possesses properties that are necessary for a glucagon antagonist to be potentially useful in the study and treatment of diabetes. PMID- 2560176 TI - Neurotensin elevates cytosolic calcium in small cell lung cancer cells. AB - The ability of neurotensin (NT) to elevate cytosolic Ca2+ in small cell lung cancer (SCLC) cells was investigated using the fluorescent Ca2+ indicator Fura 2 AM. Using SCLC cell line NCI-H345, NT elevated cytosolic Ca2+ levels in a concentration-dependent manner. Using a 10 nM dose, NT and C-terminal fragments such as NT(8-13) but not N-terminal fragments such as NT(1-8) elevated the cytosolic Ca2+ levels. Because EGTA (5 mM) did not affect the NT response, NT may cause release of Ca2+ from intracellular stores. These data indicate that SCLC NT receptors may use Ca2+ as a second messenger. PMID- 2560177 TI - Proopiomelanocortin (POMC)-related peptides in the brain of the rainbow trout, Salmo gairdneri. AB - We have investigated the presence of ACTH, alpha-MSH and beta-endorphin, three peptides which derive from the multifunctional precursor protein proopiomelanocortin (POMC) in the brain of the rainbow trout Salmo gairdneri. Using both the indirect immunofluorescence and peroxidase-antiperoxidase techniques, a discrete group of positive cells was identified in the hypothalamus, within the anterior part of the nucleus lateralis tuberis. alpha MSH-containing neurons represented the most abundant immunoreactive subpopulation. Coexistence of alpha-MSH, ACTH and beta-endorphin was observed in the lateral part of the nucleus. ACTH- and beta-endorphin-containing cells were mainly distributed in the rostral and caudal regions of the nucleus. In the medial portion of the nucleus lateralis tuberis, numerous cells were only stained for alpha-MSH. Moderate to dense plexuses of immunoreactive fibers were observed in the ventral thalamus and the floor of the hypothalamus. Some of these fibers projected towards the pituitary. The concentrations of ACTH, alpha-MSH and beta endorphin-like immunoreactivities were measured in microdissected brain regions by means of specific radioimmunoassays. Diencephalon, mesencephalon and medulla oblongata extracts gave dilution curves which were parallel to standard curves. The highest concentrations of POMC-derived peptides were found in the diencephalon (alpha-MSH: 4.28 +/- 0.43 ng/mg prot.; ACTH: 1.08 +/- 0.09 ng/mg prot.; beta-endorphin: 1.02 +/- 0.1 ng/mg prot.), while lower concentrations were detected in the mesencephalon, medulla oblongata and telencephalon. The present results demonstrate that various peptides derived from POMC coexist within the same cell bodies of the fish hypothalamus. Taken together, these data suggest that expression and processing of POMC in the fish brain is similar to that occurring in pituitary melanotrophs. PMID- 2560178 TI - Pertussis toxin inhibits neuropeptide Y-induced feeding in rats. AB - Neuropeptide Y (NPY) is the most powerful peptide drug stimulating feeding in rats. Rats with paraventricular hypothalamic (PVH) cannulae were used to investigate the mechanisms involved in NPY-induced feeding. Consistent with previous reports, injection of 2 micrograms of NPY into the PVH significantly increased the cumulative food intake over 1-, 2- and 4-hr periods. Ad lib feeding decreased significantly two days after pertussis toxin (PT) administration, but recovered to nearly normal levels on the fourth day. PT had no immediate effect on NPY-induced feeding; however, four days after PT was injected NPY (2 micrograms) did not increase the food intake compared to control. In vitro investigations showed that isoproterenol-stimulated adenylate cyclase activity in the hypothalamus of control rats was inhibited by NPY. In PT-treated rats, however, no inhibition of cAMP production was observed. These results suggest that cAMP may mediate NPY-induced feeding and that a PT-sensitive G protein may be involved in this signal transduction. PMID- 2560179 TI - Effects of CRF, AVP and opioid peptides on pituitary-adrenal responses in sheep. AB - Intravenous administration of equimolar doses of CRF (30 micrograms) and AVP (6 micrograms) to mature female sheep resulted in elevated plasma concentrations of ACTH and cortisol. Simultaneous administration of equimolar amounts of CRF and AVP resulted in a greater ACTH response compared with the sum of the responses to CRF or AVP given independently. Intravenous bolus administration of the endogenous opioid, Met-enkephalin (2.5 mg), and its potent and long-acting analogue, [D-Ala2,N-Phe4,Met(O)ol5]-enkephalin [FK33-824 (250 micrograms)], did not alter ACTH or cortisol secretion. Furthermore, naloxone, an opioid receptor antagonist given alone or concurrently with Met-enkephalin or FK33-824, was without effect. Pituitary-adrenal responses to CRF were unaltered by simultaneous administration of Met-enkephalin, FK33-824 or naloxone. These results suggest that in the sheep, opioid involvement in the tonic regulation of pituitary adrenal function is absent. However, CRF and AVP may act alone or in synergy to control the release of biologically active ACTH from the sheep pituitary gland. PMID- 2560180 TI - Physiological effects of adrenocorticotropic hormone and hydrocortisone in laying hens. AB - To study the hormonal effects on hematologic parameters as indicators of chronic stress, exogenous adrenocorticotropin (ACTH) at 6.3 or 20.0 IU/kg/day and hydrocortisone at .25 or 2.5 mg/kg/day were administered parenterally to laying hens. Both ACTH treatments induced significant (P less than .05) heterophilia, monocytosis, eosinophilia, and basophilia. Significantly elevated leucocyte counts and lymphopenia (P less than .05) were observed with the high dosage of ACTH. Both hydrocortisone-treated groups developed an absolute lymphopenia and heterophilia (P less than .05). The low dosage of hydrocortisone induced a significant (P less than .05) monocytosis; the high dosage caused significant (P less than .05) decreases in the total eosinophil and basophil counts as well as an increase in the ratio of heterophils to lymphocytes. The hemopoietic parameters, especially heterophil counts, were sensitive indicators of a hormonal stress response induced by the administration of ACTH and hydrocortisone. PMID- 2560181 TI - Passive and carrier-mediated intestinal absorption components of two angiotensin converting enzyme (ACE) inhibitor prodrugs in rats: enalapril and fosinopril. AB - The intestinal absorption mechanism of two ACE inhibitor prodrugs, enalapril and fosinopril, was investigated in rats using a single-pass perfusion method. A modified boundary layer solution was applied to determine the apparent intestinal wall permeability. The prodrug enalapril is well absorbed from rat jejunum, whereas the parent drug, enalaprilat, is poorly absorbed. The permeability of enalapril is concentration dependent and is decreased by the dipeptide Tyr-Gly and by cephradine but not by the amino acids L-leucine or L-phenylalanine, indicating a nonpassive absorption mechanism via the small peptide carrier mediated transport system. In contrast, fosinopril is readily absorbed by a concentration-independent mechanism without the involvement of the peptide carrier. PMID- 2560182 TI - Propentofylline and a hydroxy metabolite augment A2-receptor mediated cyclic AMP accumulation and attenuate A1-receptor mediated inhibition of acetylcholine release in the rat hippocampus. AB - In contrast to the adenosine receptor antagonist theophylline, the xanthine derivative propentofylline (HWA 285) is reported to protect against ischaemic cell damage. We examined the effect of propentofylline on two adenosine actions in the rat hippocampus; the A2-mediated stimulation of 3H-cAMP accumulation and the A1-mediated inhibition of 3H-ACh release. Propentofylline (0.5-1 mM) increased the adenosine (3 and 30 microM) -induced 3H-cAMP accumulation. This effect was shared by its metabolite A 72 0287. The mechanism may be a decreased inactivation of adenosine, since the effect of the stable adenosine derivative NECA was not altered. In contrast, the inhibitory effect of adenosine on evoked 3H-ACh release was inhibited by propentofylline and by its metabolite A 72 0287, but enhanced by the uptake inhibitor dipyridamole. The effect of the stable, A1 receptor selective analogue R-PIA (1 microM) was also blocked by propentofylline and by A 72 0287. In addition, propentofylline (0.5 mM) blocked the presynaptic inhibitory effect of carbachol (1 and 50 microM). Thus, propentofylline and one of its metabolites inhibits adenosine A1-receptor mediated presynaptic inhibition while it enhances adenosine A2-mediated cAMP accumulation in the rat hippocampus differently. This selectivity in action can only partly be explained by receptor subtype selectivity, and effects at sites other than the adenosine receptor are of major importance. PMID- 2560183 TI - Differences in efficacies between morphine and methadone demonstrated in the guinea pig ileum: a possible explanation for previous observations on incomplete opioid cross-tolerance. AB - We have compared the effectiveness of morphine and methadone as agonists in untreated guinea-pig longitudinal muscle preparations and after their treatment with the specific opioid receptor alkylating agent beta-chloronaltrexamine. In untreated ilea, the naloxone pA2 was 8.5 to both morphine and methadone, and their dose-response curves were parallel. After alkylation, the dose-response curve for morphine was shifted to the right with a decreased maximal effect. It was found that about 70% of the receptors were inactivated by the treatment, corresponding to a morphine receptor occupancy at IC50 of about 24%. The maximal effect of methadone was not decreased by the same beta-chloronaltrexamine treatment, indicating a much higher efficacy or receptor reserve for the drug. The possibility that differences in efficacies could account for previously reported heterogeneities in actions of opioids, such as assymmetries in cross tolerance towards morphine and methadone in experimental animal and man, is discussed. PMID- 2560184 TI - Isolation and immunological properties of adenosine kinase. AB - Bovine liver adenosine kinase is a 43 kDa protein that catalyzes the transfer of phosphate from GTP or ATP to adenosine. Its immunological properties were compared to other GTP-binding proteins of approximately 40 kDa, in particular those involved in signal transduction, such as Gs and Gi, the stimulatory and inhibitory regulatory proteins of adenylyl cyclase, Gt, from the visual excitation system, and Go, a similar protein of unknown function. Antibodies elicited in rabbits against adenosine kinase did not significantly cross-react with other guanyl nucleotide-binding proteins. Antibodies against the other GTP binding proteins did not react with adenosine kinase. Thus these GTP-binding proteins do not exhibit immunological cross-reactivity. PMID- 2560185 TI - [The importance of food and drug plant factors in the combined treatment of patients with type-II diabetes mellitus]. AB - A study was made of the effect of natural (vegetable dish) and refined (food methylcellulose--MC-100 and citrus pectin) vegetable fibers and Arfazetin (antidiabetic species, USSR) on the level of glucose, immunoreactive insulin (IRI), C-peptide, glucagon and gastrin in the blood of 41 patients with type II diabetes mellitus. These parameters were investigated on an empty stomach, 1, 2 and 4 h after breakfast I with a standard set of foodstuffs and minimum content of nutritive fibers. Similar investigations were conducted after intake of the same breakfast I in combination with various vegetable components. All the vegetable factors under study were shown to contribute to a decrease in a value of a glycemic rise noted after food intake. A vegetable dish and Arfazetin equally caused a significant rise of IRI secretion and C-peptide 60-90 min. after breakfast. MC-100 and particularly pectin decreased IRI and C-peptide secretion. There was a significant rise of gastrin secretion 1, 2 and 4 h when pectin was added to breakfast. When Arfazetin, a vegetable dish and MC-100 were added to breakfast I, a tendency to a rise of serum gastrin was observed 1 h after breakfast. The level of IRG during investigations with vegetable components did not differ from the results of investigations without these components. PMID- 2560186 TI - [The function of the hypothalamo-hypophyseal-adrenal system in Itsenko-Cushing syndrome]. AB - Altogether 28 patients with adrenocortical tumors (corticosteromas and corticoandrosteromas) were examined using a radioimmunoassay to determine the concentration of corticotropin, cortisol, blood cyclic nucleotides, the circadian rhythm and time course of corticotropin and cortisol levels against a background of insulin hypoglycemia. In patients with Itsenko-Cushing's syndrome the basel level of cortisol was notably raised, that of corticotropin was sharply decreased. The circadian rhythm of the levels of corticotropin and cortisol was disturbed, a response of the hypophyseal-adrenal system to a stressor was suppressed. The concentration of blood cyclic nucleotides was increased, and these changes were dependent on the nature of adrenal pathology. PMID- 2560188 TI - Effects of Panmede and various horse serum concentrations on the axenic cultivation of Entamoeba histolytica strains in TPS-1 medium. AB - We analyzed the influence of Panmede and horse serum concentrations on the growth of five Entamoeba histolytica strains (HK9, HM1, HM2, HM3, and HM38) axenically cultivated in TPS-1 medium. Panmede was evaluated by comparing the growth of strain HM1 in medium prepared with each of 15 Panmede lots; the yields of E. histolytica trophozoites depended on the lot quality of Panmede, and their maximal values ranged from 8 x 10(3) to 8.9 x 10(4) amoebae/ml. The growth promoting effect of eight lots of horse serum on strains HK9 and HM1 were studied using a single Panmede lot of good quality. Yields obtained with strain HK9 ranged from 8 x 10(4) to 1.8 x 10(5) amoebae/ml, whereas yields obtained with HM1 ranged from 3 x 10(4) to 1.2 x 10(5) amoebae/ml. Thus, the optimal serum concentration in TPS-1 medium that caused maximal growth of E. histolytica cultures depended on the quality of the serum lot and proved to be specific for each of the five E. histolytica strains investigated. It ranged from 18% (v/v) for strain HM2 to 28% (v/v) for strain HM1. Our results reveal that the growth of E. histolytica trophozoites in TPS-1 medium can be distinctly improved by selecting appropriate lots of Panmede and horse serum and using optimal serum concentrations. PMID- 2560187 TI - [The activation pathways and changes in the endocrine function of the testes evoked by the effect of the presence of the female]. AB - It has been shown on male mice of A/He strain that the presence of a receptive female placed behind a network partition in a cage with a male mouse caused a sharp rise on the 20-40th min. of the blood level of testosterone resulting from appropriate changes in testicular biosynthetic capacity. This rise of the level of testosterone in the male blood is realized via the transadenohypophyseal route because the antagonist of LH-RH receptor completely blocks this effect. A conclusion has been made that the activating effect of the presence of a receptive female on the blood level of testosterone is realized with the involvement of LH-RH receptors via the transadenohypophyseal route. PMID- 2560189 TI - Possible mode of action of toltrazuril: studies on two Eimeria species and mammalian and Ascaris suum enzymes. AB - The anticoccidial properties of toltrazuril in Eimeria falciformis-infected mice were potentiated by the simultaneous application of pyrimethamine, trimethoprim, or sulfadimidine. The same drugs potentiate the effect of toltrazuril by killing E. tenella schizonts in chicken kidney-cell cultures. Activities of some enzymes of the respiratory chain, such as succinate-cytochrome C reductase and NADH oxidase and succinate oxidase from mouse liver, were reduced in the presence of toltrazuril. The same effects could be observed when the activities of NADH oxidase and fumarate reductase from the nematode Ascaris suum were determined in the presence of the drug. Vertebrate enzymes involved in pyrimidine synthesis, e.g., dihydrofolate reductase from chicken liver, were also affected by toltrazuril; however, this effect was 500 times weaker than that shown by pyrimethamine. Toltrazuril also showed an inhibitory effect on the dihydroorotate cytochrome C reductase from mouse liver. Our results suggest that toltrazuril primarily affects the respiratory chain and secondarily, two enzymes involved in pyrimidine synthesis. PMID- 2560190 TI - Peptide sequencing and site-directed mutagenesis identify tyrosine-319 as the active site tyrosine of Escherichia coli DNA topoisomerase I. AB - Tyrosine 319 of E. coli topoisomerase I is shown to be the active site tyrosine that becomes covalently attached to a DNA 5' phosphoryl group during the transient breakage of a DNA internucleotide bond by the enzyme. The tyrosine was mapped by trapping the covalent complex between the DNA and DNA topoisomerase I, digesting the complex exhaustively with trypsin, and sequencing the DNA-linked tryptic peptide. Site-directed mutagenesis converting Tyr-319 to a serine or phenylalanine completely inactivates the enzyme. The structure of the enzyme and its catalysis of DNA strand breakage, passage, and rejoining are discussed in terms of the available information. PMID- 2560191 TI - The carboxyl terminal domain of Escherichia coli DNA topoisomerase I confers higher affinity to DNA. AB - Limited digestion of E. coli DNA topoisomerase I with trypsin or papain generated a DNA-binding domain of MW 14,000 corresponding to the carboxyl terminal of the enzyme. This fragment binds to single-stranded DNA agarose as tightly as the intact enzyme. It required around 400 mM NaCl for elution. A truncated topoisomerase that lacks this C-terminal domain was purified. It was eluted from the single-stranded DNA agarose column at around 150 mM NaCl. Although the truncated enzyme could relax negatively supercoiled DNA as efficiently as the intact enzyme at low ionic strength, its processivity was more sensitive to increasing salt concentration. Measurement of binding to fluorescent etheno-M13 DNA also demonstrated that the presence of the C-terminal domain confers higher affinity to DNA for the enzyme. PMID- 2560192 TI - Genetic analysis of the molecular basis for beta-adrenergic receptor subtype specificity. AB - Pharmacological analysis of ligand binding to the beta-adrenergic receptor (beta AR) has revealed the existence of two distinct receptor subtypes (beta 1 and beta 2) which are the products of different genes. The predicted amino acid sequences of the beta 1 and beta 2 receptors differ by 48%. To identify the regions of the proteins responsible for determining receptor subtype, chimeras were constructed from domains of the human beta 1 and hamster beta 2 receptors. Analysis of the ligand-binding characteristics of these hybrid receptors revealed that residues in the middle portion of the beta AR sequence, particularly around transmembrane regions 4 and 5, contribute to the subtype specific binding of agonists. Smaller molecular replacements of regions of the hamster beta 2 AR with the analogous regions from the avian beta 1 AR, however, failed to identify any single residue substitution capable of altering the subtype specificity of the receptor. These data indicate that, whereas sequences around transmembrane regions 4 and 5 may contribute to conformations which influence the ligand-binding properties of the receptor, the subtype-specific differences in amine-substituted agonist binding cannot be attributed to a single molecular interaction between the ligand and any amino acid residue which is divergent between the beta 1 and beta 2 receptors. PMID- 2560193 TI - Site-directed mutagenesis of colicin E1 provides specific attachment sites for spin labels whose spectra are sensitive to local conformation. AB - Colicin E1 is an E. coli plasmid-encoded water-soluble protein that spontaneously inserts into lipid membranes to form a voltage-gated ion channel. We have employed a novel approach in which site-directed mutagenesis is used to provide highly specific attachment points for nitroxide spin labels. A series of colicin mutants, differing only by the position of a single cysteine residue, were prepared and selectively labeled at that cysteine. A hydrophilic sequence (398 406) within the C-terminal domain of the water-soluble form of the protein was investigated and exhibited an electron paramagnetic resonance (EPR) spectral periodicity strongly suggesting an amphiphilic alpha-helix. After removal of the N-terminus of the protein with trypsin, the spectra for this sequence indicate increased label mobility and a more flexible structure. PMID- 2560194 TI - Axial ligand replacement in horse heart cytochrome c by semisynthesis. AB - Semisynthesis has been employed to replace the axial methionine in horse heart cytochrome c with histidine. The reduction potential of the His-80 protein (cyt c His-80) is 41 mV vs NHE (0.1 M phosphate; pH 7.0; 25 degrees C). The absorption spectra of oxidized and reduced cyt c-His-80 are very similar to those of the native protein in the porphyrin region, but the 695 nm band is absent in the oxidized His-80 protein. PMID- 2560195 TI - Deletions and replacements of omega loops in yeast iso-1-cytochrome c. AB - omega (omega)-loops are protein secondary structural elements having small distances between segment termini. It should be possible to delete or replace certain of these omega-loops without greatly distorting the overall structure of the remaining portion of the molecule. Functional requirements of regions of iso 1-cytochrome c from the yeast Saccharomyces cerevisiae were investigated by determining the biosynthesis and activity in vivo of mutant forms in which four different omega-loops were individually deleted, or in which one omega-loop was replaced with five different segments. Deletions encompassing amino acid positions 27-33 and 79-83 either prevented synthesis of the holoprotein, or produced highly labile iso-1-cytochromes c, whereas deletions encompassing positions 42-45 and 48-55 allowed partial synthesis and activity. These two latter regions, therefore, are not absolutely required for any biosynthetic process such as heme attachment, mitochondrial import, or for enzymatic interactions. All replacements in Loop A (residue positions 24-33) with same size (10 amino acid residues), longer (13 and 15 amino acid residues), or shorter segments (6 amino acid residues), resulted in strains having at least partial levels of iso-1-cytochrome c; however, the relative activities ranged from zero to almost the normal level. Thus, Loop A does not appear to be essential for such biosynthetic steps as heme attachment and mitochondrial import. In contrast, the full range of relative activities suggest that this region interacts with physiological partners to carry out efficient electron transport. PMID- 2560196 TI - Theoretical analysis of red cell metabolism and its interaction with membrane transport. PMID- 2560197 TI - Altered adenine nucleotide metabolism in senescent erythrocytes from the rabbit. PMID- 2560198 TI - The purification of hemoglobin solutions by heating. PMID- 2560199 TI - Oxygen sensing and erythropoietin gene regulation in a human hepatoma cell line. PMID- 2560200 TI - SC-41930 inhibits neutrophil infiltration of the cavine dermis induced by 12(R) hydroxyeicosatetraenoic acid. AB - Psoriasis is a disease state characterized by epidermal proliferation, neutrophil infiltration, along with release of the proinflammatory mediators leukotriene-B4 (LTB4) and 12(R)-hydroxyeicosatetraenoic acid [12(R)-HETE]. LTB4 and 12(R)-HETE are chemoattractant to the neutrophil, the latter approximately 1000X less potent. LTB4 and 12(R)-HETE are present in psoriatic scale, the latter in quantities so much greater than LTB4 that it is proposed as a primary mediator of neutrophil infiltration in psoriasis. 12(R)-HETE, synthesized in optically pure form by a new, shorter route, was injected into the cavine dermis. At a dose of 25 micrograms m per intradermal site, 12(R)-HETE was a significant chemoattractant to the neutrophil (as assessed by dermal myeloperoxidase levels). SC-41930, 7-[3-(4-acetyl-3-methoxy-2-propylphenoxy)-propoxy]- 3,4-dihydro-8 propyl-2H-1-benzopyran-2-carboxylic acid, given intragastrically inhibited 12(R) HETE-induced neutrophil infiltration of the cavine dermis with an ED50 value of 13.5 mg/kg. Compounds such as SC-41930 may well have utility for treating human psoriasis. PMID- 2560201 TI - Opposite effect of adrenalectomy on rat brain prostaglandin synthesis in basal conditions and in response to insulin or 2-deoxy-glucose. AB - In the present study we evaluated the role of endogenous glucocorticoids in the biosynthesis of prostaglandins (PG) in cortical brain tissue of the rat. Experiments were carried out under basal conditions and in response to insulin induced hypoglycemia and 2-deoxyglucose (2-DG) induced-cytoglucopenia. In intact rats, following hypoglycemia and cytoglucopenia, the production of brain PG was decreased. These two stress stimuli also activated adrenocortical secretory responses, as manifested by an increase in circulating ACTH and corticosterone. Bilateral adrenalectomy did not modify the brain production of PG under basal conditions. In contrast, in adrenal-ectomized rats, the biosynthesis of brain cortical PG was markedly increased in response to insulin and 2-DG. These results suggest that adrenal hormones may be involved in the modulation of cortical PG production under stressful conditions. PMID- 2560202 TI - GABAergic modulation of yawning behavior. AB - The hypothetical modulation by GABAergic neurons of yawning behavior in the rat was explored with GABA-active drugs. Gamma-acetylenic-GABA, a specific inhibitor of GABA-T, increases yawning frequency when injected at a dose of 7 mg/kg. Baclofen, a GABAB agonist (3 mg/kg), inhibits yawning completely; GABA antagonists, bicuculline and picrotoxin, at subconvulsant doses, also decrease yawning. All drugs were injected intraperitoneally with the exception of apomorphine, which was injected subcutaneously. It is suggested that GABAB receptors play a role in yawning behavior by modulating ACh release, and that GABAA receptors may modify yawning frequency by modulating inhibitory influences on ACh neurons. PMID- 2560204 TI - Enhancement of naloxone-induced analgesia by pretreatment with morphine. AB - Recently there have been demonstrations of a form of analgesia in rats that depends on the repeated administration of an opiate antagonist for its occurrence. The mechanism of this naloxone-induced analgesia (NIA) is not clear. This experiment tested the hypothesis that the relationship between behavioral effects of previous experience with opiate agonists and antagonists would be reciprocal with respect to analgesia. Consistent with such an hypothesis, prior exposure to morphine increased sensitivity to the effect of naloxone as measured by the rate of acquisition of NIA. Although receptor functions were not measured, reciprocal changes in the regulation of opiate receptors by opiate receptors by opiate agonists and antagonists may underlie the behavioral effects observed in this experiment. PMID- 2560203 TI - Neuroendocrine and beta-adrenoceptor response to chronic ethanol and aggression in rats. AB - Male rats were administered either ethanol (6-8 g/kg/day) or dextrin-maltose, an isocaloric equivalent, for two weeks prior to a 24-hour resident-intruder test. After the first 20 minutes of the aggression test residents showed a greater increase in norepinephrine than intruders (216% vs. 97%), while intruders showed a greater increase in epinephrine (394% vs. 51%) and corticosterone (338% vs. 129%) than residents. Ethanol administration increased the initial epinephrine response of intruders almost two-fold compared to dextrin-maltose intruders. After 24 hours of aggression testing plasma norepinephrine was still elevated in residents (92%) and intruders (71%), however, only intruders continued to show an elevation in plasma corticosterone (98%) and epinephrine (107%). Using a cumulative dose-response technique, the dose of isoproterenol required to produce 50% of the maximal heart rate response (ED50) increased in intruders by 108% following aggression testing with ethanol intruders showing significantly smaller mean change. The increase in ED50 was related to drug type, behavior, and plasma corticosterone and epinephrine levels. Rats treated with ethanol had a greater beta-adrenoceptor density than control rats. However, no relationship was found between receptor density and the other measures in this study. PMID- 2560205 TI - A method for exposing primates to marihuana smoke that stimulates the method used by human marihuana smokers. AB - A method was developed for exposing rhesus monkeys to marihuana smoke under conditions simulating those used by human marihuana smokers. An ADL II smoking machine was used to generate puffs of smoke of constant volume from marihuana or placebo cigarettes at a constant rate. The outlet of the smoking machine was connected by Tygon tubing to an airtight face mask covering the monkey's nose and mouth. The monkey was seated in an airtight Plexiglas chamber with only its head protruding. When a puff of smoke was generated, a vacuum was imposed on the chamber causing the monkey to inhale deeply and hold its breath. After 6 seconds, the vacuum was removed and the monkey was allowed to exhale and breathe fresh air freely until the next puff. Delivery of smoke to the monkey by this method was more efficient than allowing the monkey to voluntarily inhale the smoke. The method was used in acute dose-effect experiments and, subsequently, in long-term experiments. It may be useful with other animal species and for efficiently delivering other airborne substances of abuse. PMID- 2560206 TI - Septal alpha-noradrenergic antagonism in vivo blocks the testing-induced activation of septo-hippocampal cholinergic neurones and produces a concomitant deficit in working memory performance of mice. AB - In order to test the hypothesis that alpha-noradrenergic receptors in the septum 1) play an important functional role in the mediation of trans-synaptic control of the neurones of the cholinergic septo-hippocampal pathway and 2) produce resultant modulation of working memory performance, we have investigated the effects in vivo of the acute intraseptal injection of an alpha-antagonist, phenoxybenzamine, in mice. Neurochemical analysis was performed using measures of the kinetics of sodium-dependent high-affinity choline uptake in samples of hippocampus from injected mice and their relevant controls in both quiet conditions and immediately following selective working memory testing in an 8-arm radial maze. Results show that whereas the injection of phenoxybenzamine produces no significant alteration of the activity of the cholinergic septo-hippocampal neurones in quiet conditions, the pretrial (20 min) administration of this drug almost totally abolished the usually observed increase in hippocampal cholinergic activity induced by testing. This inhibition of cholinergic activation was associated with a parallel working memory deficit. The results provide further direct support for the hypothesis that septal noradrenergic afferents via alpha receptors mediate a phasic and net excitatory trans-synaptic influence on the cholinergic septo-hippocampal pathway during working memory testing and thereby significantly contribute to the modulation of the level of working memory performance. PMID- 2560207 TI - Effects of opiate agonists and an antagonist on food intake and brain neurotransmitters in normophagic and obese "cafeteria" rats. AB - The relationship between the effects of opiates on food intake and on central monoamines in various brain areas was investigated in normophagic and obese "cafeteria" rats. Three agonists, beta-endorphin, dynorphin, and D-Ser2-Leu-Enk Thr6 (DSLET) and an antagonist, naltrexone, were used. The three agonists enhanced feeling in normophagic rats but had different dopaminergic effects. Serotonergic metabolism increased concomitantly with the enhancement of feeding by the agonists, whereas it decreased following treatment with the antagonist naltrexone. In the cafeteria rats, although the feeding effects of dynorphin and DSLET occurred earlier, there was a complete lack of monoaminergic effects. beta Endorphin was completely devoid of effects in this model. There would, thus, appear to be a positive correlation between the behavioural effects of these opiates and serotonergic metabolism in normophagic rats, while stimulated feeding situations ("cafeteria" rats) the disruption of a monoaminergic modulation does not prohibit a direct effect on feeding. PMID- 2560209 TI - D-cycloserine, a positive modulator of the N-methyl-D-aspartate receptor, enhances performance of learning tasks in rats. AB - Glycine has recently been shown to positively modulate the N-methyl-D-aspartate (NMDA) subclass of acidic amino acid receptors which are important in neural pathways involved in learning and memory. We report that d-cycloserine (DCS), an antimycobacterial agent known to cross the blood-brain barrier, binds with high affinity to this glycine modulatory site, functions as a positive modulator, and facilitates performance of learning tasks in rats. In addition, DCS appears to be a potent cognitive enhancer at doses lower than those required for antibacterial activity. Based on these data, we propose that modulation of NMDA receptors via glycinergic mechanisms may be a means of influencing cognitive processes. PMID- 2560210 TI - Matching to sample, blood pressure and hormonal effects of chronic enalapril in baboons. AB - Sensory and behavioral performance of three normotensive and one renovascular hypertensive baboon was tested before, during and following chronic oral dosing with the angiotensin converting enzyme (ACE) inhibitor enalapril. Performance measurements during a five-color simultaneous matching to sample task were obtained during enalapril dosing of 0.18 and 0.61 mg/kg/day, and vehicle. Each dose was administered for 21 consecutive days preceded and followed by 14 baseline and recovery periods, respectively. BP from awake animals as well as serum ACE activity were measured. Systolic BPs decreased by a maximum of 6-8% (8 mmHg). ACE activity was decreased in a dose-dependent fashion by 54.01% and 81.63% for 0.18 mg/kg and 0.61 mg/kg doses, respectively. At 0.61 mg/kg, the duration of simple key-press motor behavior increased by 15% in the first week and then progressively returned to baseline levels. Systematic changes in choice reaction times or color discrimination accuracy were not observed. Although the renovascular hypertensive baboon displayed greater hypotension and ACE inhibition, behavioral effects were not significantly different from normotensive baboons. The present study extends to sensory functions the lack of adverse behavioral side-effects of enalapril. PMID- 2560208 TI - Attenuation of the biphasic effects of ethanol on avoidance extinction by RO 15 4513 in rats. AB - Ethanol had biphasic effects on jump-up avoidance extinction with low doses (1 g/kg) increasing, and high doses (2.5 g/kg) decreasing number of trials to extinction criterion. In Experiment 1 these doses of ethanol were studied alone, and in combination with RO 15-4513 (0.3, 3 or 6 mg/kg). The stimulation of responding produced by low ethanol doses was reversed by 3 and 6 mg/kg doses of RO 15-4513 which had intrinsic suppressive effects, but the depressed responding produced by higher ethanol doses was not attenuated by RO 15-4513. Experiment 2 analysed the interaction between ethanol and benzodiazepine antagonists RO 15 1788 and CGS 8216. RO 15-1788 did not have intrinsic action and did not interact with ethanol. CGS 8216 showed an intrinsic suppressive action much like RO 15 4513, and also reversed the stimulation produced by low dose ethanol, but not the effects of the high dose. Experiment 3 showed that the benzodiazepine agonist, chlordiazepoxide, had effects much like low dose ethanol which were reversed by CGS 8216 and RO 15-4513. The major conclusions were that RO 15-4513 and CGS 8216 possess inverse agonist properties which may cancel out the effects of alcohol under certain circumstances. PMID- 2560211 TI - beta-Carboline-induced seizures in mice: genetic analysis. AB - The inbred mouse strains BALB/cBy (C) and C57BL/6By (B6) differed significantly in their susceptibility to seizures induced by the benzodiazepine inverse agonist methyl beta-carboline-3-carboxylate (beta-CCM). Following a 5 mg/kg injection of beta-CCM, 74% of C (n = 35) and 13% of B6 (n = 40) mice exhibited a convulsion. No sex difference was found. Analysis of the reciprocal F1s failed to show either maternal environmental and/or heterosomal effects. A genetic analysis of the strain difference in susceptibility to beta-CCM-induced seizures using recombinant inbred strains (RIS) was performed. The strain distribution for the RIS showed a two group partition. Statistical analysis showed that, although a one-segregating-unit model could not be rejected to explain the strain difference in beta-CCM-induced seizures, some of the evidence weakened the one-segregating unit hypothesis. PMID- 2560212 TI - Hyperactivity induced by N-methyl-d-aspartate injections into nucleus accumbens: lack of evidence for mediation by dopaminergic neurons. AB - To test the hypothesis that the motor hyperactivity associated with intra accumbens injections of N-methyl-d-aspartate (NMDA) results from stimulation (direct or indirect) of nucleus accumbens dopaminergic mechanisms, the behavioral effects of intra-accumbens and intraventricular NMDA were compared to those of the prototypic dopaminergic releasing agent, amphetamine, and the competitive NMDA receptor antagonist, 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP). Drugs were injected into the right lateral ventricle, or bilaterally into the nucleus accumbens of rats. Locomotor activity was monitored electronically and by direct observation for 40 min prior to, and 1 hour after, drug treatment. Intra-accumbens injections of NMDA (0.4, 1.2 and 2.0 micrograms/side) produced dose-related increases in distance traveled, but had no significant effect on movement time or vertical movements. The NMDA-induced increase in distance traveled was temporally correlated with convulsive wild running, but not with exploratory behavior, suggesting that this increase may have been secondary to seizure-like activity. Intra-accumbens injections of amphetamine (10, 20 and 40 micrograms) or CPP (0.1 microgram) produced dose-related increases in all three measures. By the intraventricular route, the effects of NMDA were similar to those of intra-accumbens administration, whereas intraventricularly administered d-amphetamine had no effect. The behavioral effects of intra-accumbens NMDA cannot be explained by an NMDA receptor-mediated facilitation of dopaminergic neurotransmission; rather, this type of facilitation may be associated with competitive NMDA receptor antagonism. PMID- 2560213 TI - Opiate control of spontaneous locomotor activity in a urodele amphibian. AB - An intraperitoneal injection of the preferential opiate receptor agonist (+/-) bremazocine HCl given to male rough-skinned newts acutely and dose-dependently reduced their spontaneous locomotor activity. Inversely, and contrary to the situation generally observed in other vertebrates, administration of the opiate receptor antagonist naloxone HCl dose-dependently and acutely stimulated locomotion. Given at a behaviorally active dosage, naloxone counteracted the inhibitory effect of bremazocine on locomotion. The behavioral influence of the two substances was observed using two different sampling techniques (continuous recording for 3 minutes: repeated instantaneous sampling for 60 minutes). These data are discussed in view of our current knowledge on the opiate regulation of locomotor activity in vertebrates. PMID- 2560214 TI - Inhibition of transient gene expression in Chinese hamster ovary cells by triplet sensitized UV-B irradiation of transfected DNA. AB - The biological effectiveness of thymine-thymine cyclobutane dimers specifically induced by photosensitized ultraviolet-B irradiation was analyzed by host-cell reactivation of triplet-sensitized, UV-B irradiated plasmid pRSV beta gal DNA transfected into normal and repair-deficient Chinese hamster ovary cells. For comparison, pRSV beta gal DNA was also UV-C irradiated and transfected into the same cell lines. Ultraviolet endonuclease-sensitive site induction was determined after UV-C irradiation or acetophenone-sensitized UV-B irradiation of plasmid pRSV beta gal DNA. These data were used to calculate the number of cyclobutane pyrimidine dimers required to inactivate expression of the lacZ reporter gene in each irradiation condition. Transfection with UV-C-irradiated plasmid DNA resulted in a significantly greater reduction of reporter gene expression than did transfection with acetophenone-sensitized UV-B-irradiated pRSV beta gal DNA at equivalent induction of enzyme-sensitive sites. Since only a fraction of the inhibition could be accounted for by noncyclobutane dimer photoproducts, these results suggest that cytosine-containing pyrimidine cyclobutane dimers may be more effective than thymine-thymine dimers in inhibiting transient gene expression as measured in such host-cell reactivation experiments in mammalian cells. PMID- 2560215 TI - Effects of hematoporphyrin derivative on the cellular energy metabolism in the absence and presence of light. AB - Effects of Photofrin II on energy metabolism and metabolic viability were studied in a mammalian transformed cell line (BHK-21) in dark and after photo-irradiation with visible light. Cells were allowed to accumulate Photofrin by incubating for 4 h in buffer containing Photofrin (5-60 micrograms/ml). The results show that Photofrin significantly affects the cellular energy metabolism even in the absence of light; activity of cytochrome c oxidase is decreased and glucose utilization and lactate production (glycolysis) are increased. Irradiation with light resulted in a significant decrease in the activity of cytochrome c oxidase, glycolysis, ATP content, energy charge, ratios of adenine nucleotides like ATP/ADP, ATP/AMP and cell viability (dye exclusion test). Presence of inhibitors of energy metabolism, potassium cyanide (respiration) and 2-deoxyglucose (glycolysis), further enhanced the cytotoxic effects induced by hematoporphyrin derivative and light. PMID- 2560216 TI - Saline preference of cross-suckled spontaneously hypertensive and normotensive rats. AB - Two-bottle, saline preference tests were conducted on spontaneously hypertensive rats (SHR) of the Okamoto strain, normotensive Sprague-Dawley (SD) rats, SHR suckled from immediately after birth on SD foster mothers and SD suckled on SHR dams. Cross-suckling failed to alter the exaggerated saline preference of the SHR measured at 3 or 6 months of age. However, SD suckled on SHR exhibited a significantly reduced saline preference when compared to SD suckled on their natural mother. Plasma renin activity and angiotensin enzyme activity were not different between groups and saline preference did not correlate with either enzyme level. While this raises the possibility that the exaggerated saline preference of the SHR may be largely genetically determined, the role of some humoral factor in utero in the development of this appetite cannot be ruled out. The nature of the factor transferred from SHR mothers, which reduced saline preference in cross-suckled SD, remains to be determined. The possibility that this altered preference is due to a peculiarity of SHR maternal behaviour cannot be ruled out. PMID- 2560218 TI - Chromatin structure of transposon Tn903 cloned into a yeast plasmid. AB - Transposon Tn903 contains the APH gene for kanamycin resistance, which is active in yeast [A. Jimenez and J. Davies (1980) Nature (London) 287, 869-871] and is flanked by two inverted repeats (IR) 1057 bp long. When plasmid pAJ50, carrying Tn903 and the 2-microns circle origin of replication, is cloned into Saccharomyces cerevisiae, nucleosomes are assembled in vivo on the prokaryotic DNA of the transposon. Indirect end labeling revealed that three nucleosomes are preferentially positioned on symmetrical sequences from both IRs. DNase I digestion also confirmed that the chromatin structure is symmetrical in both IRs. This suggests that sequence determinants are decisive for chromatin structure in these regions. We have calculated the rotational and translational fits [H. R. Drew and C. R. Calladine (1987) J. Mol. Biol. 195, 143-173] for the Tn903 sequence and the results indicate that the nucleosome positioning on the IRs is sequence-directed. Nucleosome deposition on the APH gene also occurs, but no clear positioning exists. Some sequence preference for positioning nucleosomes on the promoter can be predicted, especially from the translational fit. Experimental data indicate, however, that nucleosomes are absent from the promoter. Therefore, chromatin can be organized on prokaryotic DNA in a manner that resembles the typical eukaryotic chromatin structure. PMID- 2560217 TI - Comparative analysis of retroviral vector expression in mouse embryonal carcinoma cells. AB - A series of replication-defective retroviral vectors were assessed for their ability to efficiently transfer functional genes into undifferentiated cells. In these vectors (designated handicapped because of a deletion of enhancer and promoter sequences in the viral long terminal repeat) transcription of inserted genes is under the control of internal promoters. Although a composite promoter composed of a mutant polyoma virus enhancer (PyF441) coupled to the herpes simplex virus thymidine kinase promoter was anticipated to function efficiently, it was found to be significantly inferior to the mouse X-chromosome phosphoglycerate kinase (pgk-1) promoter, in its ability to express the selectable neomycin phosphotransferase gene in mouse embryonal carcinoma cells. The pHMB vector, which contains the pgk-1 promoter, was shown to confer the drug resistant phenotype at high frequencies to F9 and P19 cells. This vector might prove to be of general utility for efficient gene expression in other developmental contexts. PMID- 2560219 TI - Transfer of Tn1545 and Tn916 to Clostridium acetobutylicum. AB - Tn1545, a conjugative transposon originally discovered in Streptococcus pneumoniae, has been transferred from Enterococcus faecalis and Bacillus subtilis to Clostridium acetobutylicum NCIB 8052. Transfer between different strains of C. acetobutylicum has also been observed. Insertion of Tn1545 into the C. acetobutylicum chromosome occurred at multiple sites, as shown by Southern hybridization. Although ermAM (erythromycin-resistance) was the most satisfactory marker for primary selection of transconjugants, all three Tn1545-encoded antibiotic resistance genes (aphA-3, ermAM, and tetM) were apparently expressed in C. acetobutylicum. Our results indicate that Tn1545 is potentially useful for undertaking mutagenesis and mutational cloning in this industrially important organism. Transfer of another conjugative transposon, Tn916, from E. faecalis to C. acetobutylicum NCIB 8052 was also apparently detected. Circumstantial evidence suggests that there may be a hot spot for Tn916 insertion in the C. acetobutylicum NCIB 8052 chromosome. PMID- 2560220 TI - Primary cells and established cell lines join DNA ends with the same efficiency relative to homologous recombination. AB - The joining of DNA ends was compared in the established CV1 cell line and in African green monkey kidney primary cells, from which the CV1 cell line was established. Using a linear SV40 genome that carries a terminal repeat of 526 nucleotide pairs, we measured the efficiency of circularization by end joining relative to circularization by homologous recombination between the terminal repeats. The ratios of end joining to homologous recombination were identical in the two types of cells regardless of whether the DNA ends were sticky, blunt, or mismatched. These studies demonstrate that the efficient end joining observed in established cell lines is not a peculiar adaptation to life in culture, but rather reflects a normal aspect of DNA metabolism in mammalian cells. Furthermore, the similar results in primary and established cells support the general validity of using data from studies of cultured cells to formulate models of recombinational processes in primary cells and potentially in multicellular organisms as well. PMID- 2560221 TI - Neuroregulatory effects of nicotine. AB - The impact of nicotine on the central nervous system is, in an important sense, neuroregulatory, with cascading effects on physiological and biochemical function as well as on behavioral activity. Accordingly, the neurotransmitter and neuroendocrine effects of nicotine constitute a critical part of its biological action, which includes reinforcing as well as pathophysiological consequences. This review focuses on nicotine's effects on cholinergic and non-cholinergic nicotine receptors and on the responses of catecholamines, monoamines, hypophyseal hormones, and cortisol. The contribution of critical variables, such as timing and duration of neuroregulator release and the patterns that make up the total response, is still largely unknown, particularly with regard to the effects of environmental context, history of nicotine use, and mode of administration. The evidence suggests that by altering the bioavailability of the above-listed neuroregulators, nicotine serves as a pharmacological "coping response", providing immediate though temporary improvement in affect or performance in response to environmental demands. Much of what is known to date is based on studies involving the administration of agonists and antagonists under different environmental conditions. Newer technological approaches such as autoradiography and positron emission tomography show potential for determining the neuroregulatory patterns involved and specifying nicotine's locus of action relevant to its behavioral and physiological effects. PMID- 2560222 TI - Human CRH stimulation response during acute withdrawal and after medium-term abstention from alcohol abuse. AB - We compared the baseline cortisol secretory pattern and ACTH and cortisol responses to hCRH (100 micrograms) in eight patients acutely withdrawn from ethanol and 12 patients who abstained from ethanol for two to six weeks. Acute withdrawal from ethanol was characterized by elevated baseline cortisol and blunted ACTH release after hCRH, while medium-term abstention was associated with normalized cortisol secretion but persistence of decreased ACTH output following stimulation. These findings support an altered corticotrophic CRH receptor function in detoxified sober alcoholics. The pathophysiology underlying the blunted ACTH response to hCRH in medium-term ethanol abstention appears to be different from that in acute alcohol withdrawal and hypercortisolemic depression. PMID- 2560223 TI - Opioid influences on pituitary function in sheep under basal conditions and during psychological stress. AB - The effects of intravenous injections of naloxone (2 mg/kg), morphine (0.3 mg/kg) and saline vehicle on plasma concentrations of cortisol, prolactin, vasopressin and oxytocin were assessed in sheep (N = 10) when in their social groups (basal conditions) and during a period of isolation (psychological stress). Blood samples were collected by jugular venipuncture before and during the 60-min period following drug administration. Plasma hormone concentrations were determined by radioimmunoassay. Under basal conditions, cortisol levels were increased after naloxone (36-48%), but not after morphine or saline, and concentrations of prolactin, vasopressin and oxytocin did not change. Under stress conditions, (1) cortisol concentrations were elevated throughout the 60 min sampling period after naloxone or saline but for only 20 min after morphine; maximum increases observed were 161% (naloxone), 150% (saline) and 112% (morphine); (2) prolactin levels were raised after saline (85-129%) and morphine (55-61%) but were unchanged after naloxone; (3) vasopressin concentrations decreased transiently (43%) after saline but not following naloxone or morphine; and (4) oxytocin levels did not change after any treatment. These results indicate that endogenous and exogenous opioids modulate cortisol release in nonstressed sheep, and cortisol and prolactin secretion in sheep subjected to psychological stress. The nature of the anterior pituitary responses induced, together with the absence of a discernible effect on posterior pituitary function, suggest that the central opioid systems involved are similar in sheep and primates but different from those in the rat. PMID- 2560224 TI - Influence of an uremia-associated serum factor on CNS alpha 2-adrenoceptors. AB - The action of blood serum from uremic rats and chronically hemodialyzed patients was investigated for effects on alpha 2-adrenoceptors labeled with 3H-clonidine. Compared to blood sera of rats and patients with normal kidney function, uremic serum significantly inhibited specific 3H-clonidine binding. In saturation experiments the density and affinity of alpha 2-adrenoceptors for 3H-clonidine was lowered by uremic serum. Heating, or trypsin or lipase treatment of the serum did not affect this phenomenon. The effect of the patient's serum could likewise be demonstrated after hemodialysis treatment. The presence of an allosteric regulating substance for clonidine binding to adrenoceptors could at least partially explain the altered and attenuated action of this drug in renal insufficiency. PMID- 2560225 TI - Radiographic features and prognosis in bronchioloalveolar carcinoma; a local experience. AB - The prognosis of 20 patients with histologically proven bronchioloalveolar carcinoma was assessed in the light of their radiographic features and symptoms at presentation. Radiographically, half the patients were classified as having focal disease of which 60% were focal consolidation. Other focal presentations were, a mass lesion in three cases and a single 3 cm nodule in one case. Those with diffuse disease presented with either coarse widespread nodules or diffuse alveolar opacification with or without a focal area of consolidation. All patients were symptomatic at presentation but the severity of the symptoms bore no relation to the radiological findings. Seven cases with focal disease underwent thoracotomy, five of which had evidence of hilar and mediastinal lymph node involvement. They included a case presenting with a nodule, two with a mass and four with focal consolidation. The survival following surgery was generally better with those with a mass or nodule but one case with focal consolidation remains alive 10.5 years following surgery compared with the longest survivor of 4.5 years with a mass. Both these relatively long-term survivors had no histological evidence of hilar or mediastinal lymph node involvement compared with the other surgical cases. The median survival in cases without surgery was one month. The prognosis was better in patients with focal disease where surgery was performed and we feel that a local area of consolidation should be treated as focal disease and surgical resection considered for these patients. PMID- 2560227 TI - Malignant fibrous histiocytoma of lung. PMID- 2560226 TI - Successful use of repeated transbronchial lung biopsies in a patient with multiple opportunistic infections. PMID- 2560228 TI - [A woman with a palpable breast mass]. PMID- 2560229 TI - [Pulmonary embolism caused by lipiodol: apropos of a case]. PMID- 2560230 TI - [Adrenal gland masses and Addison's disease of tuberculous origin: apropos of 2 cases]. AB - Two cases of confirmed tuberculous Addison disease, with an increase in size of the adrenal glands are presented. One patient presented also a renal adenocarcinoma; this association is very rare given the small frequency of these diseases. Based on these two cases we emphasize the utility of puncture aspiration with a fine needle (PAFN), echography and TC Scan in the study of Addison disease. PMID- 2560231 TI - [Membrane signals: role of the inositol phosphate system]. AB - Current information on cellular communications through plasmatic or synaptic mediators raises a great deal of questions about intracellular messengers diffusion and the apparent difference between the large number of intercellular mediators, or first messengers, and the few number of intracellular mediators, or second messengers. Regarding the intracellular messengers, the research is currently focused on the inositol phosphate system and especially on inositol 1,4,5 trisphosphate (IP3), which derives from the hydrolysis of a cellular membrane phosphoinositide: one of the most important function of IP3 is the release of intracellular calcium from the endoplasmic reticulum. The cell wall plays an important role in intracellular and intercellular communication as is shown by the balance between the cell-membrane phosphoinositide group and IP3 availability. Recent developments in knowledge about this inositol-phosphate system is going to show a new way of explanatory possibility about some not evident pathologies until now. The latest discovery of a specific intracellular receptor site of IP3 and the fact that some oligoelements, such as lithium, interfere with IP3 metabolism suggest exciting prospects for the treatment of pathologies resulting from these changes in receptor mechanism. PMID- 2560232 TI - 87Rb, 23Na and 31P nuclear magnetic resonance spectroscopy of the perfused rat kidney. AB - 87Rb, 23Na and 31P nuclear magnetic resonance (NMR) were used to monitor changes in renal cations and energetics during the induction of hypoxia in the isolated perfused rat kidney. The NMR-determined unidirectional Rb+ flux in normoxic kidneys was shown to be a good measure of net intracellular K+ influx in the perfused rat kidney model. The changes in 87Rb, 23Na and 31P spectra following the induction of hypoxia are consistent with hypoxic depletion of intracellular adenosine triphosphate (ATP) and a subsequent decrease in Na-K-ATPase transport activity. The exponential rate constant for 87Rb+ efflux measured during Rb+ uptake in normoxic kidneys (0.12 +/- 0.01 min-1) was not significantly different to the rate constant for 87Rb+ efflux during the induction of hypoxia (0.16 +/- 0.07 min-1). We conclude that there is no direct effect of hypoxia on renal cellular membrane integrity and that renal cell sensitivity to hypoxia is due to an inability to sustain cellular ion gradients following depletion of intracellular ATP. PMID- 2560233 TI - Renal proton magnetic resonance in experimental acute renal failure in rats. AB - Cortical, medullary and papillary T1 and T2 water proton relaxation times were measured at 37 degrees C, 20 MHz. The measurements were made using kidneys from rats affected by many forms of experimental acute renal failure (ARF), namely acute hemorrhagic hypovolemia, angiotensin II administration, antidiuretic hormone (ADH) administration, glycerol, and other nephrotoxins (gentamicin, cisplatinum, cyclosporine), renal artery occlusion for different periods of time, and ureteral ligation. From the T1 and PW (percent tissue water content) the bound water (FB) and HF (percent water bound/g solid) were calculated according to a fast proton diffusion model. In most experimental models studied, the experiments were repeated following paramagnetic enhancement with GdDTPA administration (70 mmol/kg BW). By profiling the deviations from normal, it was possible to differentiate the ischemic (shortened T1, prolonged T2), obstructive (very high T1 and T2 in both cortex and medulla) and nephrotoxic (prolonged T2) forms of ARF. Significant changes in free/bound water compartments occurred, though their biological significance is unknown. T1 and T2 ratios before and after paramagnetic enhancement correlated well with estimates of glomerular filtration rate. In the first minutes following acute hemorrhagic hypovolemia, the intrarenal water distribution remained unchanged. After GdDTPA significant water proton T1 and T2 changes characterized the immediate posthemorrhagic state similar to the effect of ADH. PMID- 2560234 TI - Intranephron distribution of purine-metabolizing enzymes in rats. AB - The activities of purine-metabolizing enzymes, 5'-nucleotidase, adenosine deaminase, and purine nucleoside phosphorylase in microdissected rat nephron segments were measured. The specific activity of 5'-nucleotidase was highest in the proximal tubules and the cortical collecting duct, but low in the glomerulus. In contrast, the highest activity of adenosine deaminase was found in the glomerulus. The distribution pattern of purine nucleoside phosphorylase was similar to that of adenosine deaminase. These results suggest that various nephron segments can form adenosine and that the glomerulus exhibits highest capacities to metabolize this nucleoside. PMID- 2560235 TI - Anemia induces 5'-nucleotidase in fibroblasts of cortical labyrinth of rat kidney. AB - We recently observed a strong increase of 5'-nucleotidase in renal fibroblasts of rats that were anemic due to an immunity against erythropoietin. In order to test if the change of 5'-nucleotidase was related with anemia, we studied the distribution of the enzyme in irradiated rats treated with phenylhydrazine. The hematocrit of these rats decreased to 15% within 4 days and erythropoietin levels were more than 200 times over controls. After 7 days a histochemical study showed that the enzymatic activity and the immunoreactivity for 5'-nucleotidase was markedly enhanced in the fibroblasts of the cortical labyrinth. There was no modification of 5'-nucleotidase in other cell types of the kidney. The 5' nucleotidase activity of renal fibroblasts in cell culture increased by 72% upon addition of 160 microM 5'-AMP to the culture medium for 8 days. We propose that anemia provokes an energy deficit in some structure in the cortical labyrinth. This might increase the concentration of 5'-AMP which would induce 5' nucleotidase. An interesting consequence of these events would be an increased production of adenosine in the direct vicinity of some of its putative targets, the glomerular arterioles and the erythropoietin-producing cells. PMID- 2560236 TI - Effect of intrarenal adenosine on urinary excretion of prostaglandins and leukotrienes in the anesthesized dog. AB - Adenosine is a renal vasoconstrictor that plays an important role in mediating renal adaptive responses to decreases in renal perfusion pressure. It is known that adenosine acts on the metabolism of arachidonic acid, but the direct repercussions of adenosine in the production of renal prostaglandins and leukotrienes have not been studied. This study was undertaken to evaluate the effect of the intrarenal infusion of adenosine upon the urinary elimination of arachidonic acid derivatives. Samples of urine were collected with lysine acetylsalicylate and determination of prostaglandins (PGs) and leukotrienes (LTs) was performed by radioimmunoassay of samples previously separated by HPLC. The infusion of adenosine decreases the urinary excretion of 6-keto-PGF1 alpha and TxB2 significantly. There was no significant change in urinary excretion of PGE2 while LTB4 and LTC4 showed a tendency to increase. These results suggest that a fall in the synthesis of PGI2 along with an increase in LTC4, which is a constrictor of mesangial cells, could be responsible for the renal vasoconstriction phase of adenosine. Therefore, it was concluded that adenosine vasoconstriction is mediated through the inhibition of the cyclo-oxygenase pathway, diminishing the synthesis of PG vasodilators. PMID- 2560237 TI - Increased cAMP levels in lymphocytes of allografted rats: possible relationship to delayed graft rejection. AB - We studied the levels of nucleotides in peripheral blood and in spleen lymphocytes obtained from allografted rats. Four types of allografts were performed by using ACI rats (RT1a) as donors and Lewis rats (RT1l) as recipients: heart, pancreas, kidney and liver. Ungrafted Lewis rats were used as controls. Rejection time without immunosuppression occurred at 6.7 +/- 0.25 days from transplant in heart-allografted, at 7.3 +/- 0.25 days in pancreas-grafted, at 6.8 +/- 0.24 days in kidney-grafted, and at 20 +/- 3 days in liver-grafted rats. Biochemical assays on lymphocytes were performed on the 7th day after operation by measuring the intracellular content of APT, ADP, AMP, cAMP, NAD+ and NADH. Our results showed that the prolonged survival of allografts in liver-grafted rats correlated with the higher content of cAMP in their lymphocytes than in those of controls. In addition, ATP and ADP levels in lymphocytes from heart-and pancreas allografted rats were significantly lower than those of lymphocytes from liver allografted rats. PMID- 2560238 TI - Specific identification of Bordetella pertussis by the polymerase chain reaction. AB - Oligonucleotide primers were used to amplify specific DNA regions of the Bordetella pertussis genome by the polymerase chain reaction. One pair of primers, PTp1/PTp2, identified a 191-bp DNA fragment located in the regulatory region of the pertussis toxin operon; a second pair of primers led to amplification of a 121-bp DNA piece located in an insertion-like element specific to B. pertussis. Both sets of primers were able to discriminate between the pathogen and related Bordetella species; they detected down to 6 bacteria and appeared suitable for routine detection of B. pertussis in clinical specimens. PMID- 2560239 TI - Glucose dehydrogenase activity in Acinetobacter species. AB - A study of D-glucose oxidation by Acinetobacter species was carried out. Glucose oxidizing strains were found distributed among almost all Acinetobacter species. 14C-glucose oxidation kinetics by non-proliferating cells with separation of oxidation products (14C-gluconate) by DEAE-cellulose paper chromatography was studied. Inhibition of glucose dehydrogenase (GDH) activity by 11 carbohydrates (mono- and disaccharides) and determination of the kinetic parameters showed that glucose oxidation was due to the action of membrane-bound GDH (inactive in vivo on disaccharides). On the basis of GDH inhibition patterns obtained, two groups were individualized. The first group of strains (identified as A. calcoaceticus, A. baumannii, A. lwoffii, A. johnsonii and Acinetobacter species 3, 9, 10 and 11) showed a greater affinity for glucose than the second group (A. haemolyticus, A. junii and Acinetobacter species 6 and 12). Restoration of GDH activity after addition of pyrroloquinoline quinone (PQQ) was studied in 187 strains previously found unable to oxidize glucose. GDH activity of 150 out of 166 strains identified as A. baumannii, A. johnsonii, A. lwoffii and Acinetobacter species 11 and 12 was restored. Eighteen of 21 strains identified as A. haemolyticus and Acinetobacter species 6 were unable to produce acid from glucose after addition of PQQ. Our results confirm that the former taxonomic scheme for the genus Acinetobacter (2 species differing only by glucose oxidation) is untenable and that, accordingly, identification of Acinetobacter strains at the species level must be performed using more modern methods, i.e. carbon source utilization tests. PMID- 2560240 TI - Alveolar macrophage dysfunctions in coal worker's pneumoconiosis. PMID- 2560241 TI - [Phyllodes breast tumors]. AB - Phyllodes tumors represent a particular class in breast pathology. They are defined as bi-tissue, fibro-epithelial tumors, similar to fibro-adenomas, but their connective tissue component is greatly predominant. They are infrequent as they represent only 3% of breast tumors. They mainly occur during the premenopausal period, since alteration of the hormonal balance may play a role. The authors feel that they represent only a type of the evolution of the fibroadenomas with a relationship, in time, following the sequence: fibroadenomaphyllodes tumor-sarcoma. The diagnosis is exclusively made by histology and there is no pathognomonic sign, whether it is clinical or mammographic. The course is essentially characterized by the possibility of local recurrences, sometimes numerous, and by the occurrence of metastases in case of malignant phyllodes tumor. These two possibilities are correlated with the histological stage of the tumor. The treatment is definitely surgical, possibly mastectomy in case of large or aggressively active tumor. PMID- 2560242 TI - [Current data on risk factors for relapse in tuberculosis]. PMID- 2560243 TI - [Computed tomography and its applications in pneumology (compared to other methods)]. PMID- 2560245 TI - [Reasons for delayed diagnosis of pulmonary tuberculosis]. AB - 100 patients at first treatment were investigated according to a complete questionnaire for explaining the causes of tardy diagnosis in pulmonary tuberculosis. The roles of the disease, patient and physician in this delay were followed. The insidious or atypical onset of the disease in 82% of these patients explains the long interval (107.2 days on the average) elapsed between the appearance of the first symptoms and the diagnosis determination. As a consequence of low cultural level, many patients cannot give a correct interpretation of the symptomatology and they seek the physician's help after 69.1 days (on the average) from the appearance of the symptoms. The medical causes are numerous and can be rapidly cured. In the dispensary of general medicine, 55% patients were examined only clinically, 45% clinically and radiologically but only 7% were subjected to the bacteriologic examination for bK and 4% were sent to the TB wards for consultation. As the registration of the patient, his admission into the hospital and the tuberculostatic treatment depend on the bK presence in sputum--sometimes waiting for the result of the culture tests--the average interval between consultation and admission is of about 34.7 days. On admission, high average values were registered: cavities of 4.2 cm in diameter, lesions of 2.8 points and bacteriologically 44 bacilli/100 fields. For improving this situation the following recommendations are made: increase of the health education on TB activity in the population, continuous education of the general practitioners in the TB diagnosis methods, use of the radiologic examination together with the bacteriologic one and the biological tests for the early detection of the pulmonary tuberculosis. PMID- 2560246 TI - [The means for evaluating and optimizing antitubercular chemotherapy as a measure to prevent and combat tuberculosis in county territory]. AB - The paper reports on 25 possible failure factors of antituberculosis chemotherapy in 262 cases recorded in 1982 and followed for 3 years. 58 of them were failures, relapses or bacteriological relapses under treatment. The failure factors analysed were subjected to a mathematical modelling method that yielded a number of codified predicting parameters. This prognostic calculation may be an efficient method for optimizing chemotherapy, reducing the failure rate. PMID- 2560244 TI - [A proposal for changing the classification of pulmonary sarcoidosis based on pulmonary functional criteria]. PMID- 2560247 TI - [Pulmonary functional criteria for evaluating the evolution of diffuse interstitial pneumopathy under treatment]. AB - It was studied the variation of the pulmonary volumes, of the gaseous transfer through the alveolocapillary membrane and of the indications of pulmonary elasticity on 40 patients with diffuse interstitial lung disease (PID), examined at intervals of 3-6 months, for 2-3 years during the treatment. The levels of the pulmonary elasticity parameters have signaled more frequently the modification (improvement or worsening) of the pulmonary functional state than those of the pulmonary volumes or of the gaseous transfer. Hence the proposal of including the tests for elastic recoil within the ensemble of pulmonary functional investigations through which are studied the effects of the treatment prescribed in PID. PMID- 2560248 TI - [Anatomicoclinical study of fatal cases of pulmonary tuberculosis at the Institute of Phthisiology in 1976-1986 (a preliminary study)]. AB - The paper reports on 95 cases of treated pulmonary and extrapulmonary tuberculosis, deceased during 10 years. In 46 of them necropsy was made. Anatomoclinical examination confirmed in 5 cases pulmonary tuberculosis and intravital neglected tuberculosis in 4 cases. Advanced forms of the disease were found at case-finding in 2/3 of the cases, this explaining the death before the treatment action. Only in 70.5% of cases the cause of death was tuberculosis, in the rest it was due to associated affection. In about half of the cases (47.4% the patients' age was over 60. PMID- 2560249 TI - [Surveillance of ventilation functions in collectives occupationally exposed to the risk of COPD]. AB - The paper presents the clinical-functional manifestations in a group of 321 workers in a carbide department of a chemical combine works, professionally exposed to powders (calcium oxide, coke dust) and irritant gases (sulphur oxides, nitrogen oxides and ozone). Data processing, recorded on a questionnaire recommended by BMRC showed an incidence of 6.8% for simple chronic bronchitis, 14% for COPD and 39.8% for the disease of the small airways. The authors support the idea of the increased efficiency of the MEF50 test in comparison with MEVS for functional surveillance and early detection of broncho-obstructive ventilatory disturbances. In agreement with other dynamical observations of the authors the subjects found to suffer from simple chronic bronchitis have, in fact, obstruction, detectable by the MEF50 test. PMID- 2560251 TI - [Organizational problems of tuberculosis treatment in a rural area]. PMID- 2560250 TI - [Should therapeutic pneumothorax really be reconsidered in the treatment of pulmonary tuberculosis? A case report]. PMID- 2560252 TI - [The evaluation of short-duration antitubercular chemotherapy (6 months) in rural areas]. PMID- 2560253 TI - [The method of organizing strictly supervised treatment in the region of the Draganesti-Olt Phthisiology Office]. PMID- 2560254 TI - [Problems of tuberculosis control activities connected with the educational and social factors of the patients]. PMID- 2560256 TI - [Medical and socio-occupational aspects of the sequelae of osteoarticular tuberculosis]. PMID- 2560255 TI - [The current situation of osteoarticular tuberculosis in the context of general tuberculosis endemic in the People's Republic of Romania]. PMID- 2560257 TI - Schizophrenia related grants--fiscal year 1988. PMID- 2560258 TI - [Prevention of postoperative thromboembolism complications by "Clexane" (low molecular weight heparin) in varicose vein surgery]. PMID- 2560259 TI - [The role of neurologic factors in Raynaud's phenomenon: epidemiologic approach]. PMID- 2560260 TI - [The use of porous hydroxyapatite and fibrin glue in periodontal surgery]. PMID- 2560261 TI - [The use of fibrin sealer (glue) and porous hydroxyapatite in the treatment of periodontal bony defects]. AB - The case report describes a method for treatment of osseous periodontal defects utilizing implantation of porous hydroxylapatite (Interpore 200) with fibrinsealer (Tisseel). Instead of the traditional flap operation-incision the Papillae Preservation Technique is used to protect the implanted material and prevent migration. The hydroxylapatite granulae were embedded in fibrinsealer before filling into the bone defects, and the flaps were fixed to the underlying tissue with fibrinsealer. One of the flaps was furthermore secured by matras suture. The clinical results after 1-year observation show healthy gingival conditions without inflammation, and no pockets being more than 3-4 mm. The radiograph shows a proper fill-up of the periodontal bone defects. PMID- 2560262 TI - Human achondroplasia: defective mitochondrial oxidative energy metabolism may produce the pathophysiology. AB - A summary is presented of previous studies by other investigators of human achondroplasia and dyschondroplastic animal models. In addition, studies previously reported from our laboratories are discussed, and they demonstrate that defective oxidative energy metabolism is present in mitochondrial preparations from achondroplastic human subjects and rabbits (ac/ac) with chondrodystrophy. The results of the studies support the hypothesis discussed fully in the manuscript that a partial defect in mitochondrial oxidative metabolism in achondroplastic subjects is expressed specifically in the growth plates of the long bones because this tissue has the lowest oxygen tension of any bodily organ undergoing active proliferation, thus leading to the achondroplastic phenotype in humans and the ac/ac rabbit. In the ac/ac rabbit phosphorylation at the cytochrome c oxidase region (site III) of the terminal respiratory system was shown to be absent in mitochondrial preparations from the livers of newborn ac/ac rabbits. Normal-appearing littermates did not exhibit the defect. Studies of mitochondrial preparations from human skin fibroblasts (grown in tissue culture) from normal human subjects and subjects with homozygous achondroplasia demonstrated that concentrations of cytochrome a3 were decreased approximately 80% in preparations from homozygous achondroplastic cells. Levels of cytochrome a3 in heterozygous achondroplastic cells were intermediate between the levels in normal cells and homozygous achondroplastic cells demonstrating the effects of gene dosage. Determination of total heme a (as the pyridine hemochromogen) in the normal and achondroplastic preparations from human subjects showed that the observed decrease in concentration of cytochrome a3 in the achondroplastic preparations was due to an absence of cytochrome a3 and not to a change in its absorbancy (extinction coefficient). PMID- 2560263 TI - [Cervical intra-epithelial neoplasia]. AB - The recommendations of proceeding in cases with CIN have significantly changed within the last years due to various reasons: Research on the biological development of CIN resulted in new concepts; as the age of patients showing-up with CIN decreased considerably, the need of more conservative diagnostic an therapeutic procedures became evident; it also was proved that human papilloma viruses contribute strongly to the etiology of CIN. So, objections against conization as the classical method of diagnosis and surgical treatment of CIN were increasingly substantiated. In order to find less invasive methods than conization, some alternatives were examined, and the combination of differential cytology, differential colposcopy and directed biopsy has proved in the meanwhile to be a valuable diagnostic alternative choice, not bearing the harmful implications of diagnostic conization. For therapy new methods have been elaborated as well, so electrocoagulation (= cauterization), cryosurgery and CO2 laser evaporization. All these methods allow a satisfactory treatment in most cases. We are recommending strongly to individualize and differentiate the procedures of diagnosis and treatment in CIN. Destructive surgery can be abandoned without taking the risk of inaccurate diagnosis or incomplete treatment. PMID- 2560264 TI - [Clinical significance of HPV infection for cervix cancer]. AB - 536 cervical biopsies with various benign and malignant epithelial changes were studied histologically and virologically with Southern Blot hybridization. In 51 cases of normal squamous epithelium HPV 16 DNA was found in four (8%) and HPV 6/11 DNA in one (2%). In 136 cases of benign acanthotic epithelium HPV 16 DNA was found in 14 (10%), HPV 6/11 DNA in six (4%), and HPV 18 DNA in two (2%). During a follow-up of four years none of these patients developed CIN. 339 further stemmed from epithelium with various degrees of CIN and squamous cell cancers. HPV 6/11 was found most often (12%) in CIN I, while in carcinomas it was found only in combination with HPV 16. In contrast, HPV 16 was found most often (43%) in invasive cancers and less often in CIN (12%). 22 CIN-I cases were available for follow-up; 18 showed regression and 4 persistency. From the 65 patients with CIN II 24 have regressed, 41 were treated by cone biopsy because of persistency. 20 of these cases harboured HPV 16/18 DNA and in two cases HPV 6/11 was found. 84 invasive cases were studied virologically. 43 patients underwent radical abdominal hysterectomy with complete pelvic and paraaortic lymphadenectomy. The surgical specimens were processed in giant sections and studied morphometrically. HPV-16-positive tumours significantly more often spread to parametrial and pelvic lymph nodes then did HPV-negative tumours. In addition the pelvic lymph nodes of 40 invasive cases were studied virologically. HPV 16 DNA was not only found in lymph node metastases of HPV-16-positive tumours, but also in lymph nodes free of metastatic disease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2560265 TI - [Endocrine active genital tumors]. AB - Endocrinically active genital tumors are indeed rare; however, their correct classification and treatment is rather difficult. If ovarian tumors coincide with abnormal hormonal activity, thorough consideration of differential diagnosis is necessary. The WHO has agreed upon a classification of endocrinically active ovarian tumors. This grouping, which is internationally accepted, is the base of the explanations, and to these, remarks about uterine HCG-producing trophoblast diseases are added. Symptomatology of endocrinically active ovarian tumors is nonuniform, depending on the tissular origin of the tumor and the pattern of its endocrine activity. Prognosis is also quite variable. This must be taken into account for therapy. E.g., operative therapy with conservation of fertility is possible--in particular in young women--with granulosa cell tumors or androblastomas; dysgerminomas are strikingly sensitive to irradiation; chorion carcinomas respond well to chemotherapy. More difficult are clear therapeutic recommendations for the very rare forms of endocrinically active genital tumors which stem from sex cords or which are composed of different components of the complex ovarian blastema. In a supplement, the relevant literature (121 quotations) is cited. PMID- 2560266 TI - The fundamental mechanism of inotropic action of digitalis. AB - Evidence from several sources, including electrophysiologic studies and cation flux and content measurements, now allows a reasonably cohesive synthesis of the molecular and cellular events that lead from inhibition of the Na pump by digitalis to the enhancement of the generation of contractile force by cardiac muscle. It is generally agreed that Na pump inhibition, with consequent elevation in [Na+]i and [Ca2+]i, underlies both the therapeutic (positive inotropic) and toxic (arrhythmic) responses of the heart, and accounts for the well-known difficulty in separating therapeutic and toxic clinical effects of digitalis glycosides. In the intact patient or experimental animal, cardiac glycosides also exert effects that are mediated through neural pathways, and these can exert important influences on the integrated response to digitalis administration. PMID- 2560267 TI - [Pancreatitis and hepatitis in a patient treated with enalapril maleate. A case report]. PMID- 2560268 TI - Detection of human papillomavirus type 52b in cervical dysplastic tissue. AB - We examined infection in the tissue of cervical dysplasia of human papillomavirus (HPV) 52b, which we recently isolated from cervical cancer. To screen a lot of samples, we used an assay system with an HPV DNA typing kit containing an HPV 33 probe that cross-hybridizes with HPV 52b DNA. DNA samples from 123 patients with cervical dysplasia were first screened with this kit. Sixteen of these appeared to be positive, and they were subjected to Southern blot hybridization with the HPV 52b probe under highly stringent conditions. Of the 16 samples, two were found to have HPV 52b genome DNA. Our results, together with the fact that HPV 52b was isolated from cervical cancer, seem to support the notion that HPV 52b infection is closely associated with the onset of cervical cancer. PMID- 2560269 TI - Radiation leukemia virus: a marker for the study of intrathymic T cell differentiation. AB - Radiation Leukemia Virus (RadLV) is a retrovirus, which displays a highly specific thymotropism; after inoculation into young C57 BL/Ka mice, active virus replication is observed only in thymocytes and thymic lymphomas develop in most treated mice. The specific interaction between RadLV and the thymus resides in the particular susceptibility to infection of the most immature cells involved in the intrathymic lymphoid differentiation pathway and of non lymphoid thymic stromal cells. Recent data obtained from 'in situ' hybridization studies for detection of viral transcripts yield more detailed informations on these intriguing interactions. PMID- 2560270 TI - Transient depletion of cortical thymocytes induced by chicken anaemia agent. AB - Chicken anaemia agent (CAA) causes severe anaemia, loss of body weight, and hypoplasia of thymus at day 14 after inoculation of one-day-old chickens. Several reports have described an enhancement of concurrent infections with f.e. Marek's disease virus, Infectious Bursal Disease virus, and Reovirus. Immunohistochemical methods were used to describe the immunopathological lesions of the thymus that probably form the basis of the immunodeficiency caused by CAA. Monoclonal antibodies and antisera against leucocytes, T lymphocytes, CD4, B lymphocytes, mononuclear phagocytes, MHC class II, and keratin were used. At day 14 after inoculation, the thymic cortex was completely depleted of thymocytes, whereas the medulla was not. T-cell areas in the spleen also lacked T lymphocytes. In contrast the cortex still contained stromal cells with MHC class II molecules and keratin. At day 21, the cortex had completely regenerated and all clinical signs of CAA infection had disappeared. Labelling experiments with BrdU in 4-week-old control chickens demonstrated that 25% of the divided cells was detected in the medulla and 75% in the cortex. The tissue tropism of CAA may, apart from the preference for rapidly dividing cells, be directed by specific cell determinants. PMID- 2560271 TI - Expression of activation antigens on thymocytes in the 'common thymocyte' stage of differentiation. AB - The authors analysed 118 monoclonal antibodies in the Activation Antigen Section of the 4th International Workshop and Conference on Human Leucocyte Differentiation Antigens for their reactivity to thymus tissue sections. Nineteen samples gave a strong labelling of cells in the subcapsular area, located in a discontinuous layer or in small foci. Positive cells also occurred incidentally in the cortex and medulla. The staining was especially prominent in thymus specimens from newborns and early childhood. This thymus staining pattern was not related to staining for activation markers in peripheral lymphoid organs or skin. After decoding and clustering, the antibodies with this thymus labelling were in various clusters including those detecting early activation antigens (CD69, Activation Inducer Molecule) and intermediate activation antigens (CD25, interleukin-2 receptor: CD30, Ki-1 group: CDw70, Ki-24 group: and CD71, transferrin receptor). But, not all antibodies in these given clusters gave thymus labelling. In two-colour analysis, cells labelled were negative for keratin and HLA-DR, and showed positivity for CD1, CD2, CD3, CD4, CD7 and CD8. This indicates that the cells are in the common thymocyte stage of intrathymic T cell differentiation, in which the majority of thymocytes in the normal cortex locate. Activation antigen expression may be related to intrathymic processing of precursor T cells with positive selection and expansion of relevant clones after surface expression of T-cell receptors. If so, the present data indicate that this process occurs already in the subcapsular area, but first after acquisition of common cortical thymocyte surface markers. PMID- 2560272 TI - The mouse T-lymphocyte-activating molecule THAM is associated to ECTO aminopeptidase activity. AB - In the mouse there are three non-CD3/T-cell receptor T cell-activating molecules, Thy-1, Ly-6 antigens, and the CD28 homologue. Using a mitogenic rat monoclonal antibody, we identified a novel thymocyte-activating molecule (THAM). Here we review the main characteristics of this heterodimeric glycoprotein with respect to its expression on lymphoid and non-lymphoid cells, and its biochemical and T cell-activating properties. THAM shares a number of structural features with surface peptidases of epithelial cell brush borders, and is in fact associated with a neutral aminopeptidase activity. The biologic significance of this observation is discussed in the context of recent data on the involvement of several ecto-enzymes in T-cell activation and differentiation. PMID- 2560273 TI - Invasive adenylate cyclase toxin of Bordetella pertussis. AB - Bordetella pertussis produces an adenylate cyclase which is a toxin. The enzyme penetrates eukaryotic cells and, upon activation by host calmodulin, generates high levels of intracellular cAMP; as a result bactericidal functions of immune effector cells are considerably impaired. The toxin is composed of a single polypeptide that possesses both the catalytic and the toxic functions. It penetrates the host cell directly from the plasma membrane and is concomitantly inactivated by a proteolytic degradation. PMID- 2560274 TI - Evolution of ATPases. PMID- 2560275 TI - Protein tyrosine dephosphorylation and signal transduction. AB - The protein tyrosine phosphatases comprise a family of enzymes that specifically dephosphorylate tyrosyl residues. Determination of the amino acid sequence of a major low molecular mass form isolated from human placenta (PTPase 1B) provided the basis for the first identification of transmembrane proteins that bear intracellular phosphatase domains. The existence of such molecules, bearing the hallmarks of receptors, raises the exciting possibility of a novel mechanism of signal transduction in which the early events involve the ligand-induced dephosphorylation of tyrosyl residues in proteins. PMID- 2560276 TI - Tissue-specific genes for respiratory proteins. AB - All but one of the mitochondrial respiratory complexes are composed of products of both the mitochondrial and the nuclear genomes. The recent isolation of cDNAs for several nuclear-encoded respiratory proteins reveals that some of them are present in at least two forms. Although some of these forms are traditional in differing somewhat in amino acid sequence, a new class, termed silent isoforms, differs in the presequence but contains identical processed proteins. What are the roles of tissue isoforms in oxidative metabolism? PMID- 2560277 TI - [Activation of the calcium channels of A-431 cells by epidermal growth factor]. AB - The cell-attached version of patch-clamp technique was used to search for calcium permeable channels in human carcinoma A-431 cells. With 100 mM CaCl2 in pipette, the inward currents were recorded having the mean unitary conductance of 2.8 pS and the reversal potential (obtained by linear extrapolation) equal to +25.5 mV. Application of the epidermal growth factor (EGF) into the bath extracellular solution produced a transient increase in probability for these channels to be open. The effect developed with a delay of about 20 seconds to last thereafter for 36 seconds (mean values). We propose that these channels mediate EGF-induced increase in concentration of cytosolic free calcium. PMID- 2560278 TI - Infection of catheterised patients: bacterial colonisation of encrusted Foley catheters shown by scanning electron microscopy. AB - The surfaces of 32 encrusted urinary catheters were examined by scanning electron microscopy to investigate the association of bacteria with the encrusting deposits. Deposits consisted of struvite crystals surrounded by aggregates of very small crystallites of hydroxyapatite. Underneath these minerals there was a layer of closely packed bacteria. Impressions of bacteria were also observed in hydroxyapatite. Crystals were often engulfed by the bacterial layer, which thus appeared to bind the crystals to each other and to the catheter surface. This thick layer of bacteria associated with crystals may protect both the bacteria from antibiotics and the crystals from acidic bladder washout solutions intended to dissolve them. Furthermore, the existence of this sessile population explains why urease-producing bacteria are not invariably detected in the urine of patients with encrusted catheters. The observation of this bacterial layer (or "biofilm") by scanning electron microscopy provided direct evidence for infection being implicated in catheter encrustation. PMID- 2560279 TI - HLA-antigen distribution in seminoma, HCG-positive seminoma and non-seminomatous tumours of the testis. AB - Histocompatibility antigens play a certain role in the development of testicular tumours. 151 patients with testicular cancer (86 non-seminomatous germ cell tumours--NSGCT--and 65 pure seminoma) were typed for the HLA-antigens of the A, B, C and DR locus. 24 patients of the pure seminoma group and 50 patients of the NSGCT group had an elevated serum HCG level preoperatively. The antigen DR-5 was elevated in the seminoma group whereas the incidence of B-13 was increased in the NSGCT group. In terms of antigen distribution HCG-positive seminoma resembles seminomatous tumours rather than NSGCT. PMID- 2560280 TI - Natural infection with the porcine respiratory coronavirus induces protective lactogenic immunity against transmissible gastroenteritis. AB - Our objective was to evaluate the level of passive protection against transmissible gastroenteritis (TGE) among 57 newborn piglets nursing from seven seropositive sows previously naturally infected with porcine respiratory coronavirus (PRCV). After challenge exposure we observed mortality rates of 44% for litters of seven PRCV-infected sows, 40% for litters of four sows orally immunized with the attenuated TGEV strain Nouzilly, and 91% for litters of seven seronegative susceptible sows. A blocking ELISA with two appropriate monoclonal antibodies distinguished serological responses of PRCV-infected sows from those of TGEV-immunized sows. The results suggest that natural infection of the sow with PRCV may induce a degree of protective lactogenic immunity against TGE. PMID- 2560281 TI - Antigenic differentiation of pestivirus strains with monoclonal antibodies against hog cholera virus. AB - Thirty-one bovine viral diarrhoea virus (BVDV) or border disease virus (BDV) strains and 94 hog cholera virus (HCV) strains were grown in cell culture, and characterized by immunostaining with 13 monoclonal antibodies (MAbs) and one polyclonal serum (PAb) against HCV. All 125 strains were recognized by the PAb. None of the BVDV or BDV strains were detected by the 13 MAbs. Seven MAbs detected all 94 HCV strains. Six other MAbs detected heterogeneity among and within HCV strains. The MAbs are useful tools in differentiating between HCV and BVDV infections in pigs, and can also be used to differentiate infections induced by HCV field strains from infections induced by the "Chinese" strain of vaccine virus. PMID- 2560282 TI - [Metastases of a malignant chemodectoma of the neck to the bone]. PMID- 2560284 TI - [Relation between echocardiographic indicators and beta 2-adrenergic receptors in essential hypertension during treatment with enalapril]. AB - A group of 22 men with essential hypertension stage I and II according to WHO, mean age 29.1 +/- 1.4 years, was divided at random into two sub-groups. Eleven patients were treated for four weeks with enalapril (E), 10 mg/day, 11 patients were given placebo (P), also for a period of four weeks. The patients were subjected to echocardiographic examination before and after four-week therapy with enalapril or placebo. In patients treated with E a significant reduction of the left ventricular wall and the interventricular septum was observed. There was also a highly significant decline of the weight of the left ventricle and the index of left ventricular mass. Enalapril led to a decline of the specific binding capacity and affinity of beta 2-adrenergic lymphocyte receptors. In the group given P no differences were found during the examination repeated after four weeks. By means of multidimensional linear regression estimates a close relationship was revealed between the difference in the weight of the left ventricle, the difference of the index of left ventricular mass and the difference of specific binding capacity and affinity of beta 2-adrenergic lymphocyte receptors. PMID- 2560283 TI - [Vascular changes in essential hypertension after treatment with enalapril]. AB - A group of patients with essential hypertension and healthy subjects from the same work place were investigated for a period of four weeks and were subjected, in addition to a basic clinical medical examination, to an ergometric examination ECHO cardiography, occlusion plethysmography, and concurrently serum levels of adrenaline (A), noradrenaline (NA), renin, cyclic adenosine monophosphate (cAMP) beta receptors before and after four weeks treatment with enalapril were assessed. The results are consistent with available data from the literature where administration of inhibitors of the angiotensin converting enzyme (ACE) caused a reduction of the peripheral vascular resistance and increased blood flow assessed by means of occlusion plethysmography. Similar results were recorded after captopril administration for a one year period, as reported by Agabiti-Rose (1). Statistically significant differences were detected between hypertonics given placebo and those given enalapril for four weeks, as regards blood flow at rest and peripheral vascular resistance at rest. There was also a significant difference when hypertonic and normotonic subjects were compared. The results are presented in tables and figures. PMID- 2560285 TI - [Prevention of thrombosis with unfractionated and low molecular weight heparin]. AB - 2 or 3 daily injections of unfractionated heparin reduce the risk of thrombosis in patients undergoing general surgery from about 39 to 4 to 5% (sodium fibrinogen-test). In patients at high risk (hip replacement, knee operations) a thrombosis incidence of about 20% is observed even under heparin prophylaxis. The clinical incidence of venous thrombosis is reduced to less than 1% in patients undergoing general surgery. Low molecular weight heparins have been investigated in a number of clinical trials. A single daily s.c. injection of low molecular weight heparin is as effective or more effective than 2 to 3 daily s.c. injections of unfractionated heparins in the prevention of thromboses in general surgery. The bleeding risk is not reduced compared to unfractionated heparin. Most frequent side effects of heparin prophylaxis are bleeding complications followed by thrombocytopenia between 1 to 2% and the rare but dangerous "white clot-syndrome" (less than 0.01%). Some patients with thrombocytopenia or white clot-syndrome can be successfully treated with a low molecular weight heparin fraction. PMID- 2560286 TI - Central bisegmentectomy of the liver: experience in 16 patients. AB - Fourteen patients with hepatocellular carcinoma and mild to moderate liver dysfunction and 2 with metachronous liver metastases from rectal cancers underwent central bisegmentectomy of the liver, i.e., en bloc removal of the left medial and right anterior segments. One patient who had undergone preoperative liver artery embolization died in the hospital. Four patients survived without relapse 60-135 months postoperatively. Eleven patients had recurrent tumors, all in the liver remnant, and 2 also had recurrent tumors in the lungs. Six patients who relapsed died 28-93 months postoperatively. Four were alive with disease at 7 89 months, 1 after a second liver resection 54 months after central bisegmentectomy. Six of the 16 patients survived for 5 years or more. The results are favorable for these advanced tumors, often in the presence of liver dysfunction. The indications, technique, and results are discussed. PMID- 2560287 TI - [Bronchopulmonary granular cell tumor]. AB - The so-called granular cell tumors, tumors of uncertain or disputed histogenesis, are generally rare, in the bronchial tree extremely rare. With regard to their localization, they are able--like a bronchogenic cancer--to cause mucus retention, bronchiectasis, recurrent pneumonias and atelectasis. A definitive diagnosis and typing of these benign tumors must be made before operation in the interest of an adequate therapy and avoiding overtreatment. PMID- 2560288 TI - [Fibrosing lung process caused by TEL mineral wool]. AB - The case of a 51-year-old man with pneumoconiosis caused by fibrous glass dust is reported. The diagnosis is confirmed by histological examination and by x-ray micro analysis using a scanning electron microscope. Pneumoconiosis following fibrous glass exposure should be taken into consideration in differential diagnosis of interstitial lung diseases. PMID- 2560290 TI - [Conjugation in group A streptococci]. AB - The paper presents data on the nature of the conjugation-like process (CLP) of genetic marker transfer on the membrane filters in group A streptococci. Studied are the CLP development depending on the duration of the donor-recipient contact and the influences of various physiological conditions on the degree of CLP manifestation and frequency. The effect of sub-inhibiting erythromycin concentrations on the frequency of the erythromycin marker transfer is first found. The original experimental evidence and literature data suggest a role played by transposing elements in CLP determination on the membrane filters in group A streptococci. PMID- 2560289 TI - [24-hour blood pressure behavior in patients with untreated and treated hypertension in comparison with normotensive patients]. AB - Blood pressure was continuously monitored over 24 h in 201 patients with mild to moderate essential hypertension using a noninvasive method. Measurements were made both before and after 6 months of antihypertensive treatment and the data were compared to results from 100 normotensive patients. The frequency with which blood pressure values above 140/90 mm Hg occurred during the 24-h period proved to be the most reliable parameter for distinguishing between hypertensive and normotensive profiles. The blood pressures of all patients could be normalized (less than 140/90 mm Hg) on single or combined drug therapy as assessed by casual measurement. However, significant differences were observed between the 24-h profiles of the treated patients and the control group. The mean 24-h blood pressure, the mean day and nighttime blood pressures, the mean hourly pressure, and the frequency of increased blood pressure values were all significantly higher in the patients on medication as compared to the normotensive controls. This would suggest that normotension, as defined by the control group, cannot be attained with antihypertensive medication. In conclusion, 24-h continuous blood pressure monitoring allows a better evaluation of blood pressure profiles and consequently, will be of greater value in assessing cardiovascular risk than occasional random measurements. PMID- 2560291 TI - Distribution of M protein and nucleocapsid protein of vesicular stomatitis virus in infected cell plasma membranes. AB - The association of M protein and the nucleocapsid (N) protein of vesicular stomatitis virus (VSV) with the cytoplasmic surface of plasma membranes prepared from infected cells was examined by double label immunofluorescence. M protein in association with the cytoplasmic surface of the plasma membrane was distributed in two distinct labeling patterns. Punctate labeling of M protein in the plasma membrane was observed in association with corresponding labeling for the nucleocapsid protein. Diffusely labeled M protein was distributed in areas of the plasma membrane that were devoid of any detectable labeling for the nucleocapsid protein. Similar results were obtained with two different cell types at 4 h and later times postinfection. The diffuse label for M protein was present in membranes prepared from cells infected with a temperature-sensitive M protein mutant at the nonpermissive temperature, but neither the punctate label for M protein nor labeling for the nucleocapsid protein was observed. Upon shift to permissive temperature, both the punctate label for M protein and labeling for the nucleocapsid protein began to reappear in membranes prepared from cells infected with the M protein mutant. These results indicate that M protein can associate with the plasma membrane without prior binding to nucleocapsids and that association of functional M protein with the plasma membrane is required for the stable association of nucleocapsids with the membrane during the process of viral budding. PMID- 2560292 TI - Naturally occurring serotype 2/subgroup II rotavirus reassortants in northern Brazil. AB - Nine serotype 2 human rotavirus strains were isolated in a community-based longitudinal study in Northern Brazil. Five of these strains had a 'long' RNA electrophoretic pattern and all five strains were determined to belong to subgroup II by ELISA assay, in contrast to properties common to serotype 2 human rotaviruses previously characterized. Hybridization studies of one of these unusual strains with 32P-labelled mRNAs derived from the prototype human strains Wa (serotype 1, subgroup II) and S2 (serotype 2, subgroup I) suggested that it was generated by a reassortment event in nature, in which a subgroup II, 'long' electropherotype rotavirus exchanged its serotype-specific gene and gene number 10 for the equivalent genes from a serotype 2, 'short' electropherotype virus. PMID- 2560293 TI - Antigenic variation of foot-and-mouth disease virus of serotype C during propagation in the field is mainly restricted to only one structural protein (VP1). AB - The primary structure of VP3, VP2 and VP4 capsid protein genes has been determined for six epizootiologically-related foot-and-mouth disease virus (FMDV) isolates of serotype C1, two of which presented immunogenic differences as determined by a cross-protection assay. The results obtained have been compared with those previously reported for the corresponding VP1 genes Martinez et al. (1988) Gene 62, 75-84. High rates of fixation of mutations have been estimated for the four capsid protein genes that ranged from 3.9 X 10(-4) to 4.5 X 10(-3) substitutions per nucleotide per year, with the highest values corresponding to VP1. Despite this genetic heterogeneity most of the amino acid exchanges are within the VP1 protein. Of the fourteen amino acid substitutions one was located in VP2 and two in VP3. Five out of the eleven amino acid exchanges that affected VP1 were located within residues 138-149, part of a main immunogenic site in FMDV. These results show that in the course of a foot-and-mouth disease outbreak, immunologically relevant amino acid substitutions occur mainly in viral capsid protein VP1. PMID- 2560294 TI - Restricted replication of human adenovirus type 5 in mouse cell lines. AB - Infection of mouse BALB/c 3T3 cells by adenovirus 5 resulted in at least 1000 fold lowered yields of virus compared to human cells. The molecular basis of this restriction was analysed at the level of viral gene expression. Steady-state levels of viral DNA and RNA were greatly reduced in infected mouse, compared to human cells. Both early region 1A (E1A) and E1B mRNAs were decreased in mouse cells and their protein products were barely detectable by metabolic labelling of infected cells. The E2A-72 kDa protein and the hexon protein were detected by metabolic labelling, and immunocytochemical analysis showed that they were correctly located in nuclei of infected mouse cells. Only a minor proportion of infected mouse 3T3 cells expressed the E2A-72 kDa or hexon proteins. Low yields of virus were obtained by infection of SV40 transformed BALB/c 3T3 cells showing that SV40 does not provide a helper function for adenovirus 5 growth in this cell system. PMID- 2560295 TI - Severity of disease induced by a pancreatropic Coxsackie B4 virus correlates with the H-2Kq locus of the major histocompatibility complex. AB - Coxsackie B viruses are known etiological agents of pancreatic diseases, including diabetes. The pathogenesis of these infections is influenced by both host and viral factors. In this report, we examined whether the outcome of Coxsackie B4 virus infection is dependent on the genes within the major histocompatibility complex (MHC). We generated a pancreatic variant, CB4-V and established an animal model system of pancreatitis with concurrent hypoglycemia in mice. Infection of various B10 H-2 congenic strains of mice revealed that the development of hypoglycemia with accompanying pancreatitis was independent of the MHC haplotype. However, the severity of the disease as monitored by the extent and duration of hypoglycemia and by mortality rate was found to be associated with the H-2 haplotype, specifically the H-2Kq locus. Pancreatic damage induced by CB4-V appeared to be both immune-mediated and viral-mediated. Histological examination of pancreatic tissue from infected B10 H-2 congenic mice revealed an association between acute destruction of the exocrine pancreas and lymphocytic infiltration. This infiltration may correlate with immune-mediated destruction of the infected pancreatic tissue. Since preferential replication of CB4-V was not observed in the most susceptible B10 mouse strain, direct viral destruction may not be the major mechanism of pancreatic injury. PMID- 2560296 TI - Wilms--a man, a syndrome. AB - A biography is presented in commemoration of the 70th anniversary of the death of Max Wilms, whose name is almost exclusively known in association with the mixed tissue tumours of the kidney. Describing his curriculum vitae, we intended to place particular emphasis on the significance of his surgical work at the University of Heidelberg. PMID- 2560298 TI - [Reduction of bony periodontal pockets with hydroxyapatite]. AB - If you can't reduce a periodontal bone defect of more than 6 mm with a conservative therapy you can open up the pocket with a marginal incision. The granulation tissue must be removed carefully, open up the surrounding spongiosa and clean the tooth from calculus. HA-Granula is filled into the pocket but don't overfill it. Use Matrix-sutures to sew the wound very dense against saliva. A final splint stabilizes the tooth. PMID- 2560297 TI - Suppression of cerebrospinal fluid (CSF) production by a Na+/K+ pump inhibitor extracted from human cerebrospinal fluid. AB - A low molecular weight compound (about 600 daltons) extracted from human cerebrospinal fluid, and sensitive to proteolytic enzymes, has been shown to mimic the specific inhibitory effects of cardiac glycosides on the Na+/K+ pump of erythrocytes. The compound, which was labelled CSF-inhibitor (CSF-I) and reconstituted in artificial rabbit CSF, was used to study its effects on the rate of CSF formation in rabbits. Three groups of adult New Zealand white rabbits of either sex, anesthetized with ketamine and xylazine, and artificially respired were subjected to ventriculocisternal perfusions. Baseline rates of CSF formation were obtained during the first 2 hours of perfusion when plain rabbit CSF was used as the perfusate. Thereafter, the animals were perfused for an additional 2 to 3 hours with either plain rabbits CSF (controls), rabbit CSF containing 10(-6) M ouabain (group 2) or CSF-I (group 3). The rate of CSF formation in control animals was observed to gradually decline with time (about 7.5% in 4 hours). The addition of ouabain to the perfusate caused an abrupt and marked 43% decline in the rate of formation while the addition of CSF-I resulted in a 57% decline. These results suggest that the peptide CSF-I which is present in human CSF may act as an endogenous regulator of CSF production. PMID- 2560299 TI - [Borderlines between success and failure in use of hydroxyapatite in daily practice]. PMID- 2560300 TI - [The importance of growth factors in endocrinology and oncology]. AB - Investigations dealing with the interrelations between peripheral hormones like proteohormones and steroid hormones and local hormones like growth factors in the target cells let us have the knack of the mechanism of action of peripheral hormones. The best known growth factors (EGF, TGF alpha, TGF beta, PDGF, IGF I, IGF II) are mentioned. The interrelation between growth factor and peripheral hormones in the target cell is demonstrated on the example of Estradiol and EGF. The practical consequences of growth factors are pointed out. PMID- 2560301 TI - [Adjuvant radioactive gold therapy of primary stage I c ovarian cancer]. AB - 47 patients with primary ovarian cancer stage I c have been treated following radical operation additionally with an intraperitoneal instillation of 5.5 GBq radioactive gold colloid. 5-year survival rate was 87 per cent. Severe complications have not been observed. PMID- 2560302 TI - [Herpes simplex virus infection in the first weeks of life]. AB - Most neonatal Herpes-simplex-virus infections are the result of virus acquisition during passage through the birth canal. Transplacental transmission of HSV is seldom. The infection is mostly localized at the skin, eyes and central nervous system. The disseminated HSV-infection represents as the most severe clinical picture. It is reported about meningoencephalitis caused by HSV in 3 infants in the first weeks of life. The therapeutic success of antiviral chemotherapeutics depends very much on their application at an early stage of the disease, which can be recognized by cerebral sonography and detection of IgM-antibodies against HSV. PMID- 2560303 TI - [Ablative hormone therapy and chemotherapy of metastatic breast cancer]. AB - Case report of a 45-year-old woman with breast cancer who underwent mastectomy with axillary lymphonodectomy and radiation therapy. Two years after primary therapy liver metastases have been proved by sonography, liver scintigraphy and computerized tomography scanning. A nearly complete remission was achieved by ablative hormone therapy and CMF-chemotherapy. PMID- 2560304 TI - [Malignant transformation of fibroadenoma of the breast]. AB - Report of a 46 year old woman with a carcinoma, developing in a fibroadenoma in the right breast, as a picture of multicentric growth of carcinoma of the breast. PMID- 2560305 TI - Congenital brain tumours: diagnostic and therapeutic approach. With a report of 3 cases. AB - Three cases of congenital brain tumours are reported, which presented with signs of increased intracranial pressure soon after birth, giving rise to the suspicion of a cerebral haemorrhage or of hydrocephalus. Correct diagnosis of tumour was established by computerised tomography, which additionally demonstrated a concomitant haematoma in each case. Two of the newborns had a primarily fatal course, with no specific treatment of the tumours being feasible. The remaining child underwent two operations, experiencing an unhindered neurologic and mental development thereafter. The histologic diagnoses were spongioblastoma and medulloblastoma in the first two cases, and ganglioneuroblastoma in the last. A conspicuous clinicopathologic feature of this neuroblastoma was the marked change in its growth pattern, revealing a higher degree of histologic differentiation and less malignant biological behaviour when tumour regrowth occurred. The diagnostic, therapeutic, and prognostic implications for this special clinicopathologic condition are discussed, with a review of the relevant literature. PMID- 2560306 TI - Efficiency of the radio-chemo-surgical. Treatment in cerebral astrocytomas. AB - This paper reports on the evaluation of 29 patients with I-st grade, 35 patients with II-nd grade astrocytomas and 145 patients with malignant astrocytomas (gr. III or IV) with a postoperative Karnofski index of minimum 60, treated with wide surgical resections, chemotherapy (CCNU 130 mg/m2 body surface) and irradiation with 60Co or conventional roentgen-therapy, in doses of 50-60 Gy, fractionated applied. The control group was formed by patients treated surgically and by postoperative irradiation: 5 with I-st and II-nd grade astrocytomas and 30 with malignant ones. Comparison of the results shows a median survival of 71.5 weeks for I-st grade, 66.5 weeks for II-nd grade and 44 weeks for high malignant astrocytomas treated only with CCNU, as against 150, 119 and respectively 73 weeks when chemotherapy was associated with irradiation with 60Co. The statistical differences between the results of radiochemotherapy and the cobalt therapy applied alone were also very significant. The long-term survival: at 5 years for I-st and II-nd degree and at 1 and 3 year for malignant astrocytomas revealed the same superiority of the complex associated treatment (radio-chemo surgical). No significant differences in survival were recorded in relation to age, sex and site in surgically treatable cases. However the infiltrative character of the tumors significantly aggravate the prognosis. It is concluded that association of chemotherapy with cobalt-therapy postoperatively brings about a statistical significant increase in survival as against postoperative chemotherapy or irradiation applied alone. PMID- 2560307 TI - [The postnatal ontogenetic characteristics of adrenal function in female water voles Arvicola terrestris with different fur colors]. AB - Studies have been made of the effect of colour mutation on the development of the adrenals in female aquatic voles. Colour mutation in homozygote results in retarded growth rate of the whole body and the adrenals, as well as in postnatal changes in the dynamics of corticosteron content in the blood. Obviously, the effect of colour mutation on plasma concentrations of corticosteron is realised mainly via extra-adrenal mechanisms since no differences were found in corticosteron production by the adrenals and in the adrenal reaction to ACTH (5 U/g of the adrenals) in vitro in growing aquatic voles. During postnatal life, mutant black females exhibited lower sensitivity of the adrenals to the inhibitory influence of estradiol (40 ng/100 mg of the adrenals) as compared to wild phenotype animals. PMID- 2560308 TI - [The functional similarity of vertebrate rhodopsin and of a photosensitive pigment from the unicellular flagellate alga Chlamydomonas reinhardtii]. AB - Photoreceptor membranes in the bovine retina may be substituted in reconstructed systems by pigment fractions from Chlamydomonas reinhardtii which account for GTP dependent activation of cGMP-phosphodiesterase. Fractions from carotinoid-less mutant did not exhibit this capacity. The results obtained reveal significant structural and functional similarity between rhodopsin-like pigment of Ch. reinhardtii and rhodopsin from vertebrates. PMID- 2560309 TI - Comparison of the characteristics of infiltrates in skin and nerve granulomas of leprosy. AB - The characteristics of infiltrates in the dermal and neural granulomas from the same leprosy patients were compared by preparing a single cell suspension. Skin and nerve biopsies from 10 patients, 5 with tuberculoid and 5 with lepromatous leprosy were analysed. The granulomas contained lymphocytes and macrophages. Lymphocytes were the predominant infiltrating cells in the tuberculoid dermal and neural granulomas. A high proportion of lymphocytes in both the skin and nerve granulomas in these cases were activated T cells as they formed rosettes with sheep erythrocytes and expressed HLA-DR antigens. In contrast, lepromatous dermal and neural granulomas contained very few of these lymphocytes. Dermal and neural granulomas from both the types of leprosy contained mature macrophages as they were esterase positive, did not exhibit peroxidase activity and expressed HLA-DR antigens. These macrophages did not possess C3 surface receptors either. These findings suggest that the infiltrates in the skin and nerve granulomas of a given type of leprosy have similar characteristics. PMID- 2560310 TI - Isolation and characterization of cells in granulomas of nerves of leprosy patients. AB - Single cell suspension from the granulomas in nerves of leprosy patients were prepared for an in vitro study of the properties of infiltrating cells. Nerve biopsies from 17 patients with tuberculoid (n = 9) and lepromatous (n = 8) leprosy cases were analysed. The granulomas were found to contain lymphocytes and macrophages. Lymphocytes were the predominant infiltrating cells in the tuberculoid nerves. In contrast, lepromatous nerves contained very few of these cells. The majority of lymphocytes in tuberculoid granulomas were activated T cells as they formed rosettes with sheep erythrocytes, exhibited esterase dots in the cytoplasm and expressed HLA-DR antigens. A small proportion of the lymphocytes also formed rosettes with EAC. Most macrophages from both the granulomas were mature macrophages as they were esterase positive, did not exhibit peroxidase activity and expressed HLA-DR antigens. The macrophages did not possess C3 surface receptors. PMID- 2560311 TI - Relationship between the extent of intraductal component and that of the invasive component in ductal carcinomas of the breast. AB - To clarify the relationship between the extent of the intraductal component and the invasive component in cases of invasive ductal carcinoma (IDC) of the breast, we divided 87 such cases into two groups, Group I in which the intraductal component extended for less than 10 mm (36 cases) and Group II in which the intraductal component extended for 10 mm or more (51 cases). On histological slides, there was an association between the extent of the intraductal component and the pattern of invasion. The majority (80.6%) of Group I IDC cases showed an invasive component composed of one invasive nodule, whereas 62.7% of Group II IDC cases showed an invasive component composed of two or more such nodules. On gross and microscopical examination, Group II IDC showed a significantly larger mean tumor size than Group I IDC (23.4 +/- 8.9 mm vs. 18.3 +/- 6.6 mm, P less than 0.002 & 24.1 +/- 14.1 mm vs. 17.1 +/- 6.5 mm, P less than 0.002). A similar result was obtained by clinical examination (41.8 +/- 17.0 mm vs. 29.5 +/- 11.6 mm, P less than 0.0001). These results suggest that Group II ductal carcinomas may frequently develop multiple stromal invasion, resulting suddenly in a sizeable breast mass perceived by the patient. PMID- 2560312 TI - Neuroectodermal differentiation in "extraskeletal Ewing's sarcoma". AB - A small round cell tumor of soft tissue arising in the retroperitoneum of an 18 year-old woman is reported. The light and electron microscopic features of the tumor were mostly indistinguishable from Ewing's sarcoma of the bone and in some parts from differentiating neuroblastoma with mature neurons and Schwann cells. An immunohistochemical study using anti-NSE antibody showed many widespread NSE positive cells lying singly and, more often, in clusters including the undifferentiated areas. The tumor was not anatomically related to the paravertebral sympathetic ganglia or adrenal glands. The present findings strongly suggest that certain extraskeletal Ewing's sarcomas are very primitive neuroectodermal tumors. PMID- 2560313 TI - [Effect of dexamethasone on Coxsackie B-2 virus-infected rat beating heart cells in culture]. AB - The effects of dexamethasone (Dex) on cultured rat beating heart cells infected with 100 TCID-50 Coxsackie virus B-2 (CB2V) were observed. The beating % began to decrease in the infected group 2 or 3 d post-challenge. Meanwhile, the cytopathic effect (CPE) appeared rapidly from 1+ to 3+. In the infected and Dex-treated group 1 h after inoculation, the beating % and CPE in the whole flask were significantly higher and less, respectively, than that in the group infected (P less than 0.05) at the same intervals. At 5 d after challenge, the beating % in the whole flask was significantly higher than that in the infected group. The cardiac enzyme-aspartate aminotransferase (AST) in the infected group was higher than that in the infected and Dex-treated group (P less than 0.01) through 3-5 d post-challenge. Moreover, the AST levels in these 2 groups were also higher than that in the uninfected group, Dex control group at the same intervals (P less than 0.01). Ultrastructural findings were parallel to the results of CPE through 1-5 d post-challenge in these 4 groups. It is suggested that the protective effect of Dex on cultured beating heart cells infected with CB2V occurred in the early stages after infection. It is surmised that steroids can probably save the lives of patients with severe myocarditis if the conventional therapy for protecting the myocardium and improving immunity were administered together. PMID- 2560314 TI - [Photodynamic action of hypocrellin A on hepatoma cell mitochondria and microsomes]. AB - Hypocrellin A (HA), a perylene quinone derivative, is a new photosensitizer extracted from Hypocrella bambusae (B et Br) Sace. A high voltage sodium lamp was used as the light source; the illumination intensity was 105 mW/cm2. After HA 25 micrograms/ml and illumination for 10 min, mitochondrial ATPase and microsomal G 6-Pase of hepatoma cells were intensively inhibited, but mitochondrial MAO was not affected. Sulfhydryl contents of the mitochondrial and microsomal membrane proteins were significantly reduced. Lipid peroxidation of mitochondrial and microsomal membrane lipids were greatly enhanced. It is concluded that mitochondria and microsomes are the sensitive targets in cells with respect to HA photosensitization. PMID- 2560315 TI - Effect of recombinant human alpha A interferon on Visna virus multiplication in acutely infected ovine cells. AB - We have studied the effects of recombinant human alpha A interferon (IFN) on the multiplication of Visna retrovirus in ovine cells. Pretreatment with IFN followed by continuous IFN treatment, drastically blocked the Visna virus induced cytopathogenic effect and the emergency of neoformed virions. Thus, virus yield and virus dependent reverse transcriptase activity were highly reduced in supernatant fluids. All these effects were accompanied by a strong inhibition of viral protein synthesis. In the light of these data, human IFN action on Visna retrovirus seems to be determined by a mechanism of action distinct from that described in the case of other Retroviridae. PMID- 2560316 TI - Enalapril decreases plasma prolactin levels in hypertensive patients. AB - Angiotensin II stimulates prolactin release both in vivo in the rat and in vitro in anterior pituitary cell cultures. Moreover, angiotensin II binding sites have been identified in pituitary lactotrophs and it has been shown that angiotensin converting enzyme (ACE) is present in rat anterior pituitary. We studied the effect of enalapril, a potent converting enzyme inhibitor, on baseline prolactin levels in nine hypertensive postmenopausal women. The results indicate that 15 day inhibition of ACE by enalapril reduced prolactinaemia, suggesting that angiotensin II plays a role in the control of prolactin secretion in hypertensives. PMID- 2560317 TI - Effects of cancer chemotherapy on the relation between serum levels of soluble interleukin-2 receptors and CD4/CD8 ratio in patients with solid neoplasms. AB - The clinical significance of sIL-2R in solid tumors has still to be clarified. To further define the biological role of sIL-2R in cancer and their relation to chemotherapy, we have measured serum levels of sIL-2R and CD4/CD8 ratio in 45 patients with limited or metastatic solid tumor, 28 of whom had never received chemotherapy, whereas the other 17 had been previously treated with chemotherapy. sIL-2R were significantly higher in metastatic cancer patients than in the non metastatic ones, while no difference was seen between patients treated and untreated with chemotherapy. Within the untreated group, sIL-2R mean values were significantly higher in patients with low CD4/CD8 ratio than in those with the normal one, while an opposite behavior was seen in patients previously treated with chemotherapy. The present study shows that cancer chemotherapy influences the release of sIL-2R and its relation to T lymphocyte subpopulations. PMID- 2560318 TI - [Wilms tumor. Evaluation of the important prognosis factors in 22 treated cases]. AB - Twenty-two cases of treated Wilms tumor with more than two years of follow-up were classified according to Beckwith (favorable and unfavorable histology) and to Jereb & Sandstedt (histologic types I, II and III). The authors found a good correlation between histologic types I and II and prognosis (72% of the children with type I were free of tumor and of type II only 25%), but in type III no correlation was found between the histologic grading and prognosis, possibly due to the small number of cases observed. The three cases classified as unfavorable histology corresponded to histologic type III. All the children less than two years old at the time of diagnosis were free of tumor, in this group six out of seven cases were of histologic type I. In the older age group only one third of the cases were free of tumor. Otherwise, 70% of the cases with unfavorable prognosis were four or more years old, but only one had unfavorable histology. The present paper shows that the age and the histologic type determined according to the criteria of Jereb & Sandstedt are important data in the prognostic evaluation of Wilm's tumor. PMID- 2560319 TI - [Effectiveness of radiotherapy in non-resectable cavernous hemangioma of the liver]. AB - In the management of the unresectable cavernous hemangioma of the liver, the Radiotherapy has an important role. This paper reports the experience of Radiotherapy department of Hospital Santa Maria. The authors review retrospectively the literature concerning techniques, results and complications of this therapy and analyse other possible attitudes in treatment of these lesions. PMID- 2560320 TI - [Current possibilities of surgical treatment of hepatocellular carcinoma. Apropos of a clinical case]. AB - The recent improvement in surgical of therapy of carriers of hepatocellular carcinoma is due to both an early diagnosis of the illness and in the evaluation of set of prognostic and resectability factors. On describing a clinical case of a hepatocellular carcinoma localized on the left lobe of a healthy liver, the importance of the analysis of those factors of prognostic and resectable evaluation is shown up, compared to those in our case. Finally, a methodology is proposed for a diagnostic tracing in accordance with the present possibilities of curable and palliative therapeutics. PMID- 2560321 TI - A study on the antiviral action of a polyphenolic complex isolated from the medicinal plant Geranium sanguineum L. VIII. Inhibitory effect on the reproduction of herpes simplex virus type 1. AB - The antiviral activity of a polyphenolic complex, isolated from a Bulgarian medicinal plant Geranium sanguineum L., on the reproduction of herpes simplex virus type 1 was studied. The substance inhibited the intracellular stages of viral replication and exhibited a virucidal effect. PMID- 2560322 TI - [The effect of antiviral substances on herpes simplex virus type 1 in cell culture]. AB - Comparative studies were carried out of the antiviral efficacy of 11 substances on the replication of Herpes simplex virus type 1 (strain DA), including compounds with selective antiherpes action--bromovinyldeoxyuridine, virazole, phosphonoformic acid, acycloguanosine, citarabine, vidarabine, iododeoxyuridine, and some nonspecific viral inhibitors with antiherpes effect as distamycin A, tunicamycin and deoxyglucose. The antiherpes activity of the substances was tested in two cell systems: cell strain of human embryonic skin-muscle fibroblasts and a cell line of green monkey kidneys. The dose-response curves and the 50% effective dose were defined for each compound by using a microtissue plate test system. Differences in the values of the 50% effective doses of iododeoxyuridine, phosphonoformic acid and distamycin A for the two types of cultures were found out. Bromovinyldeoxyuridine exhibited the highest antiherpes activity in both types of cells, and it was comparatively more active than citarabine, acycloguanosine or iododeoxyuridine, followed by vidarabine and virazole. The results obtained emphasized the necessity of specifying the experimental conditions for comparative studies. PMID- 2560323 TI - Parameters of the wave M in 398 patients with tetanic syndrome proved with positive ischemic or hyperventilation tests. AB - 398 patients with clinical signs of tetanic syndrome were examined. Their ischemic and/or hyperventilation tests were positive. The parameters of the wave M were analysed i.e. latency, amplitude, duration, shape and strength of stimulus. The signs of higher irritability of the peripheral nervous system were found out in women; they diminished during age. The ratio between men and women was 1:3.6. The significant differences were determined between the groups of negative and positive tests. In the group of positive tests we noticed the dependence of the wave M on the positivity of ischemic, hyperventilation tests, on amount of multiplets during the ischemic test, on the seasons of year and on the course of the period of years (1970-1974 and 1975 to 1978). The highest irritability was determined in the subgroup of hyperventilation tetany and in the subgroup with cramps provoked during ischemic tests. PMID- 2560324 TI - Potent and specific inhibitors of mammalian phosphoribosylpyrophosphate (PRPP) synthetase. AB - The monophosphates of the exocyclic amino ribonucleosides, 4-amino- and 4-methoxy 8-(D-ribofuranosylamino)pyrimido[5,4-d]pyrimidine, are potent and specific inhibitors of human erythrocyte and B-lymphoblast PRPP synthetase. The inhibition by MRPP monophosphate is competitive (Ki = 35 microM with the PRPP synthetase cofactor, Pi (Km = 2 mM). The nucleosides are phosphorylated to the active metabolite by adenosine kinase and these nucleoside monophosphates accumulate in the cell. beta-ARPP is a substrate, albeit poor, for adenosine deaminase and solutions of the beta-anomer of this nucleoside and its monophosphate anomerize over time to give alpha- and beta-mixtures. beta-MRPP is more resistant to adenosine deaminase and anomerization of the nucleoside and its monophosphate is negligible. The effect of treatment of cells with the nucleosides is a time dependent and nearly universal reduction in the nucleotide content which appears to result from a reduction in the availability of PRPP for dependent metabolic pathways. In studies with the WI-L2 lymphoblasts, some of these pathways, de novo and salvage (hypoxanthine and guanine) synthesis of purine nucleotides, are more sensitive to a restriction of PRPP availability than others, i.e. de novo pyrimidine synthesis. The nucleosides have shown promise as therapeutic agents in a mouse leukemia evaluation system but may also have future use in unravelling the complex regulation of PRPP synthetase and the dependent nucleotide synthesis pathways. PMID- 2560325 TI - Biochemical pharmacology and antitumor properties of 4-amino-8-[beta-D ribofuranosylamino]pyrimido-[5,4-d]pyrimidine. AB - 1. APP is activated by adenosine kinase to its 5'-phosphate (APP-MP). 2. APP-MP inhibits PRPP synthetase, and depletes cellular PRPP and purine and pyrimidine nucleotides. 3. APP inhibits synthesis of DNA and RNA, and blocks cells in G1 phase of the cell cycle. 4. APP retains full activity against MDR cells. 5. APP is equally active against quiescent and proliferating CHO cells. 6. APP has only weak activity against L1210 leukemia in vivo, but has substantial activity against mammary carcinoma 16/c. 7. In vitro, APP has a relatively high ratio of solid tumor: leukemia activity. PMID- 2560326 TI - Nuclear polyphosphoinositides during cell growth and differentiation. AB - When highly purified nuclei of Swiss mouse 3T3 cells are incubated with gamma-32P ATP, radioactivity is incorporated into phosphatidic acid and the two polyphosphoinositol lipids, phosphatidylinositol(4)P and (4,5)P2. If the cells are pre-treated with IGF-I, the incorporation into the polyphosphoinositides is decreased. This effect is maximal by 2 min, is transient in that it disappeared by 1 hr, and is increased markedly by the co-addition of bombesin, even though bombesin alone has no effect. Friend cells exhibit a related phenomenon in that the labelling of PIP2 in isolated nuclei is increased by conditions which cause erythroid differentiation (DMSO addition). We suggest that some aspect of nuclear polyphosphoinositide metabolism is modified when the nucleus is induced to divide or to differentiate, and that this change in inositide metabolism is a very early event in the sequence leading to cell division or differentiation. PMID- 2560327 TI - 6-Phosphofructo-2-kinase and fructose-2,6-bisphosphatase from Saccharomyces cerevisiae. AB - In permeabilized yeast cells 6-phosphofructo-2-kinase and fructose-2,6 bisphosphatase are studied during growth. It is shown that in yeast at least two fructose 2,6-bisphosphate degrading enzyme activities occur, differing in pH profile and in their substrate affinities. The activities of 6-phosphofructo-2 kinase and of fructose-2,6-bisphosphatases drop in the exponential and the transition phase while the activity of the alkaline phosphatases steadily increases. In the stationary phase the activities of 6-phosphofructo-2-kinase and of the low Km fructose-2,6-bisphosphatase increase again. Yeast 6-phosphofructo-2 kinase and fructose-2,6-bisphosphatase were purified and separated from each other. The purified 6-phosphofructo-2-kinase was found to exhibit a very high specific activity (1.3 U/mg). The enzyme is efficiently inhibited by ATP. The ATP inhibition is most pronounced at low concentrations of magnesium and fructose-6 phosphate. Phosphoenolpyruvate and sn-glycerol 3-phosphate are inhibitors of the enzyme. The high-affinity yeast fructose-2,6-bisphosphatase releases inorganic phosphate from the 2-position of fructose 2,6-bisphosphate. It displays hyperbolic kinetics towards fructose 2,6-bisphosphate (Km = 0.3 microM) and is strongly inhibited by fructose 6-phosphate. The inhibition is counteracted by sn glycerol 3-phosphate. The enzyme is shown to be inactivated by cAMP-dependent phosphorylation and reactivated by the action of protein phosphatase 2A. PMID- 2560328 TI - Mechanisms of short-term (second range) regulation of the activities of enzymes of lipid and phospholipid metabolism in secretory cells. AB - In isolated guinea pig parotid gland lobules the activities of the following enzymes were measured 30 sec after stimulation with either 2 X 10(-5) M isoproterenol or 10(-5) M carbachol: glycerol kinase (EC 2.7.1.30), glycerolphosphate acyltransferase (EC 2.3.1.15), lysophosphatidate acyltransferase (EC 2.3.1.51), phosphatidate phosphohydrolase (EC 3.1.3.4), diacylglycerol acyltransferase (EC 2.3.1.20), diacylglycerol kinase (EC 2.7.1.107), and CDP-diacylglycerol synthetase (EC 2.7.7.41). Lyso-phosphatidate acyltransferase, diacylglycerol kinase, and diacylglycerol acyltransferase exhibited significant increases following stimulation by both types of agonists. Stimulation of the activities of these three enzymes occurred also following in vitro incubation with the catalytic subunit of cAMP-dependent protein kinase or a Ca2+/calmodulin-dependent protein kinase II. These effects could be reversed by incubation with various protein phosphatases. When cells were first stimulated with either type of agonist, subsequent incubation with protein kinases was almost ineffective. Activation by the two types of protein kinases was not additive, indicating that they activate by phosphorylating identical sites on the enzyme proteins. The other enzymes examined showed no or only minor changes and their activities could not be affected by in vitro incubation with the two types of protein kinases. The results explain the rapid changes in acyl-group transfer from acyl-CoA to neutral lipids observed previously during the first seconds after stimulation of guinea pig parotid gland lobules with isoproterenol or carbachol (1). An analysis of a potential role of lipocortins for the regulation of phosphoinositide-specific phospholipases C reveals that these proteins do indeed inhibit these enzymes, but that this inhibition results from a calcium dependent interaction of the lipocortins with the phospholipid substrate. A physiological role of lipocortins for the regulation of phospholipases is doubtful. PMID- 2560329 TI - NAD synthesis by erythrocytes in phosphoribosylpyrophosphate synthetase (PRPPs) superactivity. PMID- 2560330 TI - ITP-pyrophosphohydrolase and purine metabolism in human erythrocytes. PMID- 2560331 TI - Cerebrospinal fluid cyclic nucleotide alterations in the Lesch-Nyhan syndrome. PMID- 2560332 TI - Activity and effect of purine metabolizing enzymes in the digestive tract. PMID- 2560333 TI - AMP catabolism in primary rat cardiomyocyte cultures. PMID- 2560334 TI - Purine catabolic enzymes in human synovial fluids. PMID- 2560335 TI - Isozyme shift of adenylosuccinate synthase in rat and human neoplasms. PMID- 2560336 TI - Effect of alterations of the ATG translation start codon of the APRT gene. PMID- 2560337 TI - Deduced amino acid sequence from human phosphoribosylpyrophosphate synthetase subunit II cDNA. PMID- 2560338 TI - Analysis of molecular structure of rat phosphoribosylpyrophosphate synthetase genes. PMID- 2560339 TI - Transfer of human HPRT gene sequences into neuronal cells by a herpes simplex virus derived vector. PMID- 2560340 TI - ATP-dependent mineralization of hyaline articular cartilage matrix vesicles. PMID- 2560341 TI - Increased purine nucleotide degradation in the central nervous system (CNS) in PRPP synthetase superactivity. PMID- 2560342 TI - Further evidence for a 'new' purine defect, inosine triphosphate (ITP) pyrophosphohydrolase deficiency. PMID- 2560343 TI - The biosynthesis and actions of prostaglandins in the renal collecting tubule and thick ascending limb. AB - PGE2 formed by renal collecting tubules is an important factor in regulating NaCl and water reabsorption in the collecting tubule and medullary thick ascending limb. PGE2 appears to act, depending on its ambient concentration, via several different receptors present in these renal epithelia to modulate cAMP turnover in both positive and negative directions. These putative PGE receptors form a family of receptors, all coupled to G proteins, and this family of PGE receptors is homologous to the adrenergic receptor family. PMID- 2560345 TI - The development of new antileukotriene drugs: specific leukotriene D4 antagonists and 5-lipoxygenase inhibitors. PMID- 2560344 TI - Peptide hormones, cytosolic calcium and renal epithelial response. AB - We have reviewed the evidence that a number of hormones interact with renal tubular epithelial cells. The evidence suggests that in the mammalian renal tubule bradykinin and parathyroid hormone interact with cell surface receptors to initiate the hydrolysis of PIP2 leading to the formation of I 1,4,5P3 and diacylglycerol in the distal and proximal tubule, respectively. The activation of this second messenger system leads to the mobilization of Ca2+ from intracellular stores. Vasopressin does not activate this second messenger system in mammalian renal epithelial cells, and we cannot demonstrate I 1,4,5P3 formation and Ca2+ mobilization either in the rabbit papillary collecting tubules or in MDCK cells. There is evidence emerging, but not discussed here, that angiotensin II may also mediate some of its effects on the mammalian proximal tubule via the inositol polyphosphate second messenger system. PMID- 2560346 TI - Endocrine abnormalities in patients with endstage renal failure. AB - Data presented in this study suggest existence of hyperendorphinism in uraemic patients. This hyperendorphinism may be regarded both as a primary beneficial compensatory mechanism counteracting disturbances of the internal environment, while causing secondary harmful side effects, which contribute to the uraemic state. Erythropoietin treatment of uraemic, haemodialyzed patients is followed by marked endocrine alterations (suppression of plasma levels of STH, ACTH, prolactin, glucagon, aldosterone, cortisol and plasma renin activity, elevation of plasma insulin and atrial natriuretic levels, lack of influence on plasma PTH, CT and AVP). It remains to be clarified whether the erythropoietin induced endocrine alterations are due to correction of the existing anaemia or reflect a specific effect of this hormone. PMID- 2560347 TI - [Granular-cell tumor of the larynx]. AB - Laryngeal granular cell tumors are rare. The interest of this subject is their peculiar histogenesis, because there is no general agreement on it. The AA. rapport one case in a woman aged 32, and review closely the related literature. PMID- 2560348 TI - Determination of total (Na+ + K+)-ATPase activity of isolated or cultured cells. AB - The aim of this work was to determine if the total (Na+ + K+)-ATPase of the plasma membrane of a cell population could be assayed without cell homogenization and partial purification of the enzyme. Several types of intact cells that were placed in an assay medium containing MgATP, Na+, and K+ hydrolyzed little or none of the added ATP. When the cells were pretreated with the ionophore alamethicin and then placed in the assay medium, they exhibited an ouabain-sensitive (Na+ + K+)-ATPase activity that increased and reached a limiting value with increasing alamethicin concentration. Since alamethicin did not increase the activity of the purified membrane-bound (Na+ + K+)-ATPase, its effects on the intact cells are probably due to the formation of large channels within the plasma membrane that allow the free access of the components of the assay medium to the intracellular domains of (Na+ + K+)-ATPase. Utilizing whole cells treated with alamethicin, total (Na+ + K+)-ATPase activity was determined in clonal pheochromocytoma cells (PC12), neuroblastoma x glioma hybrid cells (NG108-15), and myocytes isolated from adult and neonatal rat hearts. With the use of this whole-cell assay, the ouabain sensitivities of the enzymes in adult and neonatal rat heart myocytes were determined and found to be the same as those that have been determined with the use of partially purified enzymes. PMID- 2560349 TI - Bidirectional sequencing of supercoiled plasmid DNA. AB - In this paper we show that restriction DNA fragments can prime DNA synthesis of a homologous supercoiled plasmid DNA. Using the dideoxyribonucleotide chain terminator method, newly synthesized truncated chains can be detached from the primers by restriction enzyme digestion. Therefore, by choosing DNA fragments flanked by two different restriction enzymes sites, nucleotide sequence information can be simultaneously obtained on both regions of the DNA surrounding the restriction fragment. The advantage of this sequencing approach over current methods is that no prior knowledge of the primary sequence is needed to find the nucleotide sequence of a given DNA fragment. Thus, synthetic primers are not required and internal sequences of a given clone can be easily accessed without the need of fragmenting the original construct. The method has been used with rapid plasmid preparations, thus considerable time and effort can be saved in the gathering of nucleotide sequence information. PMID- 2560350 TI - A competitive enzyme-linked immunosorbent assay: applications in the assay of peptides, steroids, and cyclic nucleotides. AB - Indirect competitive enzyme-linked immunosorbent assays (ELISAs) that can be used to quantify several types of small, bioactive molecules, including peptides, steroids, and cyclic nucleotides, are described. The assays require no special expertise to perform, and the sensitivities are very high, equally or exceeding what is commonly achieved in radioimmunoassay (RIA). The molecule to be assayed or a synthetic derivative is coupled to a protein carrier (= conjugate). The conjugate is adsorbed to the wells of a microtiter plate where it is bound by antibody in inverse proportion to free hapten in a sample or standard. Bound antibody is then quantified with enzyme-labeled anti-immunoglobulin and appropriate substrate. The assay of peptides is illustrated for the sulfated cholecystokinin octapeptide, in which an ED50 of 20 fmol (2 x 10(-10) M in 100 microliters assay volume) is attained. The ED50's and slopes of the dose-response curves in the steroid and cyclic nucleotide ELISAs are compared with those parameters obtained earlier by RIA using the same antisera. This comparison indicates that a steroid, ecdysone, can be quantified with no apparent participation of the bridging group of the conjugate in the competitive assay. Furthermore, the ED50's in the ecdysone assays (ecdysone 2 beta, 3 beta, 14 alpha, 22R, 25-pentahydroxy-5 beta-cholest-7-en-6-one, 7.7 fmol; 20 hydroxyecdysone, 16 fmol) are 19- to 38-fold lower for ELISA than for RIA. In the cyclic nucleotide assay, the bridge of a cAMP conjugate (homologous with the bridge of the immunogen) decreases the slope of the dose-response curve. This effect is minimized by the use of short incubations with anti-cAMP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2560351 TI - [Effect of estroprogestative drugs on the secretion of corticoadrenal hormones]. AB - Estro-progestative drugs (EPD) have been used for many years and multiple secondary effects have been reported concerning mainly the glucidic and lipidic metabolisms. Effects of EPD on steroid metabolism have also been described and the authors present a review of the literature on this subject. Particularly it appears that EPD affect: the glucocorticoid metabolism, with augmentation of total cortisol although not in its free-form (augmentation of transcortine), the adrenal androgens metabolism, with diminution of their circulating rates, the aldosterone metabolism with augmentation of its secretion. These biological constatations could help explaining some clinical features occurring with administration of EPD: diminution of hirsutism and/or acne, augmentation of body weight, appearance of hypertension. PMID- 2560352 TI - Heavy metals in sewage sludge utilized in agriculture. AB - To evaluate the fertilizing and polluting potential of sewage sludge and to establish whether the latter limits the former or not, sludge must be characterized, application rates chosen, and sludge effects monitored. This work emphasizes that chemical analysis aimed at total metal quantification cannot by itself ensure reliable answers about the agricultural value of sludge. The EEC directives about heavy metals in sewage sludge provide the total content limitation of each element without taking into consideration the respective fractions of metals present in their free state constituting the chemical state most directly related to sanitary risk. To evaluate the role of the Environmental Impact Assessment (EIA) in a such disposal system, a scheme is proposed. PMID- 2560353 TI - [Primary cytomegalovirus infection and duodenal ulcer in a non-immunosuppressed adult]. PMID- 2560354 TI - New modified method for the surgical treatment of syndactyly. AB - We have been using Marumo's method for the surgical treatment of syndactyly. However, this method presents difficulties with incision designs, which we solved by devising a modified method. This method uses an angulated rectangular dorsal flap and a triangular miniflap made at the end of the dorsal flap. By rotating this miniflap proximally, the skin defect produced at the dorsal base of the digit after removal of the angulated dorsal rectangular flap becomes triangular. This triangular skin defect can be adequately covered by the triangular flap produced at the ventral side. In the article we describe this new operative method in detail and present cases to which the method has been applied. PMID- 2560355 TI - Familial Poland's syndrome. AB - Absence of the right pectoralis major muscle and hypoplasia of the ipsilateral breast was observed in 3 females and hypoplasia or agenesis of the right pectoralis major muscle in 2 males of the same family. One member of this family, a 17-year-old girl, underwent a right latissimus dorsi muscle flap, placement of silicone-gel breast implant, and a right infraclavicular natural Y prosthesis. Although none of our 5 patients from this family had congenital upper limb abnormalities, we believe that they still qualify as having Poland's syndrome. This would distinguish them as being the fourteenth known family reported in the world literature with this diagnosis. A brief review of the history as well as hypotheses of etiological mechanisms are presented. PMID- 2560356 TI - The preoperative use of extra-tissue expander for syndactyly. AB - To our knowledge, this is the first report of a method of stretching the interdigital skin of syndactyly by means of a pincer. The pincer, a U-shaped metal spring plate with a sponge cushion, is applied before the operation for about two months; pressure is placed against both sides of the interdigital fusion to expand the skin and to develop hollows. Use of the pincer facilitates a syndactyly operation by allowing defect coverage with a local flap only. Four patients successfully treated by this method are reported. The method has the great advantage of not requiring skin grafts, which often have unfavorable results both functionally and cosmetically. PMID- 2560357 TI - A survey of the feline leukaemia virus in Singapore. AB - In this survey for the presence of the feline leukaemia virus (FeLV) in the Singapore domestic cat population, the sera of two different groups of unvaccinated mainly short haired cats which were over 6 months old were sampled. The FeLV enzyme-linked immunoadsorbent assay (ELISA) diagnostic test kit was used to detect the presence of the FeLV group specific (gs) antigens in the blood of cats. Of the 345 clinically healthy cats surveyed, 34 sera (9.9%) were found to be positive and of the group of 123 cats with clinical signs such as chronic wasting, marked by anaemia, anorexia and lethargy, 33 sera (26.8%) were found to be positive. From the time of diagnosis of a viraemia, 70% of cats will die within 20 months. The results are therefore indicative that annually a small proportion of cats in the local environment will die from a FeLV infection. This survey reflects the natural distribution of an infectious oncovirus in a susceptible host population which is unaffected by any control programme to interfere with the normal sequence of events of host virus interactions. PMID- 2560358 TI - IgA nephritis: a review of the pathogenetic mechanisms and the rationale for therapy. AB - Various pathogenetic mechanisms are involved in IgA nephritis: immunological; platelet, coagulation and vascular injury; mesangial cell proliferation and contractility; hypertension; glomerular hyperperfusion and tubulo-interstitial injury. It is now possible to identify the subgroup of patients with IgA nephritis who have adverse prognostic features and may develop progressive glomerular scarring with renal failure. These features are proteinuria greater than 1 gm/day, non-selective proteinuria, glomerulosclerosis, hypertension, crescents and medial hyperplasia of blood vessels on renal biopsy. Controlled trials involving cyclophosphamide, anti-platelet agent (dipyridamole) and low dose warfarin; prednisolone; angiotensin converting enzyme inhibitors and eicosapentanoic acid (fish oil) appear promising. Currently, patients with bad prognostic indices in the Department are offered entry into an ongoing controlled trial of dipyridamole and low dose (anti-thrombotic dose) warfarin. Those patients with nephrotic syndrome especially with selective proteinuria are treated with a course of prednisolone and failing that, cyclophosphamide. It is important to maintain adequate blood pressure control among hypertensive patients as uncontrolled hypertension can lead to accelerated renal failure. With the onset of even mild renal impairment, dietary protein restriction should be recommended as this will help to decrease the rate of renal deterioration due to glomerular hyperfusion. PMID- 2560359 TI - [Intrapenile neurotransmission. Physiologic data and practical consequences]. AB - This paper reviews the intrapenile control of erection and the principal pathophysiological and therapeutic implications (especially intracavernous injections) of these physiological data. The regulation of flaccidity is fairly well known. This principally results from a continuous adrenergic discharge responsible, via activation of the alpha receptors, for tonic contraction of the smooth muscle fibres (SMF) of the corpora cavernosa (CC), preventing blood from entering the spaces of the CC. The complementary role of serotonin, histamine, prostaglandins (PG) F1 and F2 alpha and neuropeptide Y is still unclear. The regulation of erection is less well understood. The principal phenomenon is probably inhibition of the anti-erectile alpha adrenergic tone, allowing relaxation of the SMF and congestion of the areolae of the CC, influx of arterial blood and occlusion of the venous exits. However, an additional relaxing nervous stimulation may also be required. The modulation of anti-erectile adrenergic activity could be the result of Vasoactive-Intestinal-Polypeptide which, however, is unable to induce complete erection on its own, and/or PGE1. Acetylcholine, whose role is still unclear, stimulation of beta adrenergic receptors and presynaptic alpha-2 receptors, histamine, a relaxant factor of endothelial origin and possibly other neurotransmitters as yet unidentified, may also be involved. PMID- 2560360 TI - [Exeresis of residual masses of germinal tumors of the testis after chemotherapy]. AB - Between 1978 and 1988, amongst the 184 patients treated at the Hopital du Val de Grace for a testicular germ cell tumour, 47 patients underwent resection of residual masses after chemotherapy: 27 patients were classified as stage II and 20 were classified as stage III. The chemotherapy, administered for 3 to 6 cycles, used three types of protocols: VAB 6, PVeBV or BEP. Resection of residual masses was only undertaken after return to normal of the biological markers with persistently abnormal medical imaging. Histology of the testicular tumour revealed 3 seminomas and 44 non-seminomatous germ cell tumours. The histology of the residual masses after retroperitoneal lymphadenectomy (42 operations), thoracotomy (9 operations) or craniotomy (1 operation) revealed 22 cases of fibrosis or necrosis (47%), 10 cases of teratoma (21%), 8 cases of cancer (17%) and 7 normal cases (15%). The progression towards fibrosis, necrosis or nature teratoma is synonymous with cure, but the persistence of cancer cells corresponds to a very poor prognosis even after salvage treatment. PMID- 2560361 TI - Influence of pH, nutrient availability, and growth rate on amine production by Bacteroides fragilis and Clostridium perfringens. AB - Dimethylamine, methylamine, propylamine, and pyrrolidine were the major amines formed by Bacteroides fragilis NCDO 2217 during the active phase of growth in batch culture. Production of these metabolites was strongly pH dependent and was optimal under acidic conditions (pH 6.0). Low pH also favored the formation of pyrrolidine, cadaverine, and dimethylamine by Clostridium perfringens C523, but the reverse was the case with putrescine, butylamine, and propylamine, where production was maximal at neutral pH. B. fragilis was grown in continuous culture under either starch or casein limitation. Amine formation was influenced by carbohydrate availability and was greatest when the bacteria were grown at high growth rates (dilution rate, 0.20/h) under starch limitation, where they constituted about 18% of the total fermentation products measured. Amine production was optimal and increased concomitantly with growth rate when C. perfringens was grown in glucose-limited continuous culture. Under conditions of high growth rate and glucose limitation, amines accounted for approximately 27% of the fermentation products measured. When glucose in the feed medium was increased from 5 to 15 g/liter, amine production was repressed, and under these nutritional conditions the growth rate had little effect on the process. PMID- 2560362 TI - Comparison of ozone inactivation, in flowing water, of hepatitis A virus, poliovirus 1, and indicator organisms. AB - In steadily flowing water at 20 degrees C and pH 7, five organisms had the following order of resistance to ozone (at constant levels of ozone): poliovirus 1 (PV1) less than Escherichia coli less than hepatitis A virus (HAV) less than Legionella pneumophila serogroup 6 less than Bacillus subtilis spores. The tests were repeated at 10 degrees C with HAV, PV1, and E. coli. Ozone inactivation of HAV and E. coli was faster at 10 degrees C than at 20 degrees C. At 20 degrees C, 0.25 to 0.38 mg of O3 per liter was required for complete inactivation of HAV but only 0.13 mg of O3 per liter was required for complete inactivation of PV1. PMID- 2560363 TI - [Malignant retroperitoneal fibrous histiocytoma of the inflammatory type. Apropos of a case]. AB - A case of inflammatory malignant fibrous histiocytoma is described herein. We discuss its clinical features, therapeutic possibilities and diagnosis, highlighting the usefulness of magnetic resonance imaging as well as the importance of correct anatomopathological diagnosis. PMID- 2560364 TI - Functional and metabolic effects of bucladesine (dibutyryl cyclic AMP) in the working rat heart preparation: comparison with dopamine. AB - The effects of bucladesine (10(-4) and 3 x 10(-4) M) on cardiac function and energy metabolism were investigated in comparison with those of dopamine (3 x 10( 6) and 10(-5) M) in the working rat heart preparation. Bucladesine dose dependently increased systolic pressure, cardiac work and coronary flow, but scarcely changed heart rate or cardiac output at 78 cm H2O perfusion pressure. Dopamine increased systolic pressure to a similar extent as bucladesine and also increased heart rate, cardiac work and coronary flow, but did not change cardiac output. Neither drug caused any changes in energy metabolites. At reduced perfusion pressure (15 cm H2O), bucladesine increased systolic pressure, coronary flow and cardiac output, but caused a decrease (at 10(-4) M) or a negligible change (at 3 x 10(-4) M) in heart rate. At 10(-4) M, there was only a slight change in energy metabolism, but at 3 x 10(-4) M, anaerobic metabolism was enhanced. Dopamine decreased systolic pressure and coronary flow and increased heart rate and cardiac work. Cardiac output increased transiently and, thereafter, decreased. Dopamine also enhanced anaerobic metabolism at both doses. The results suggest that both drugs exert similar effects at normal perfusion pressure, but after reduction of perfusion pressure, some differences occur between the two drugs, presumably due to acceleration of the heart rate and subsequent increase in cardiac work by dopamine. PMID- 2560365 TI - Mechanism of nafazatrom-induced inhibition of rat uterus contractions in vitro. AB - Nafazatrom inhibits, in a dose-dependent way, the amplitude and frequency of the rhythmic contractions induced by oxytocin (4 mU/ml), as well as the methacholine (10(-5) M)- and CaCl2 (10 mM)-induced contractions, and the phasic response to KCl (60 mM); similarly, it inhibits the tonic contraction induced by KCl (60 mM) and oxytocin (10 mU/ml). A single concentration of nafazatrom (10(-4) M) also inhibits the uterine contractions caused by carbachol (10(-4) M) and prostaglandin F2 alpha (10(-6) M). CaCl2 (0.1-10 mM) only partially reverses the inhibition produced by nafazatrom on the KCl-induced tonic contractions. Bay K 8644 (3 x 10(-10)-3 x 10(-7) M) reverses the inhibition by 10(-4) M but not by 3 x 10(-4) M of nafazatrom on the CaCl2-induced contractions. Nafazatrom also inhibits, in a dose-dependent way, the calmodulin-dependent phosphodiesterase activity. Our results would suggest that, independently of its 5-lipoxygenase blocking activity, nafazatrom inhibits the contractions of the rat uterus by inhibiting the calmodulin activity and, presumably, by reducing the influx of extracellular calcium and/or the mobilization of intracellular calcium. PMID- 2560366 TI - Characterization of tight binding of isoprenaline and terbutaline to beta adrenoceptors in lung membranes. AB - Preincubation of guinea-pig lung membranes with the beta-agonist isoprenaline or with the partial beta-agonist terbutaline, followed by repeated washing, causes a 35% decrease in the number of beta-adrenoceptors (R). This decrease corresponds to the tight binding of the agonist (H) to R as a result of the formation of a complex between H.R. and the stimulatory guanine nucleotide regulatory component (Ns). Tight binding of the agonist is dependent on the presence of magnesium ions and is reversed by GTP. However, the stability of the H.R.Ns-complex is limited even in the absence of GTP. Under these conditions, the complex is more stable when induced by isoprenaline than by terbutaline. When the alkylating reagent N ethylmaleimide is included in the agonist preincubation step, tight binding of isoprenaline and terbutaline is increased to about 55% of the receptor population and is no longer dependent on the presence of magnesium ions. This value is in good agreement with the percentage of agonist high affinity sites, determined by isoprenaline/[3H]-dihydroalprenolol competition binding studies. PMID- 2560367 TI - Assessment of occlusal tooth wear in vervet monkeys by reflex microscopy. AB - A biostereometric method was used to assess the tooth wear of monkeys fed either on one of two Western-type diets, which differed in their refined carbohydrate, fat and fibre contents, or on a combination of the two diets. Certain patterns of wear as a function of diet were observed but these could not be explained adequately in terms of diet roughness alone. PMID- 2560368 TI - Ultrastructural morphology of secretory granules in pleomorphic adenoma of human parotid and submandibular salivary glands. AB - Secretory granules were found in some of the cells lining the lumina of typical epithelial structures in pleomorphic adenomas. They were small, of varied electron density, and were mostly unipartite, sometimes bipartite and occasionally tripartite. They most closely resembled secretory granules of intercalary ductal cells of normal salivary glands. PMID- 2560369 TI - Lectin binding in tissues from hydatidiform mole, invasive mole and choriocarcinoma to concanavalin-A, wheat germ agglutinin and peanut agglutinin. AB - A light microscopic analysis of lectin receptors in normal placenta and trophoblastic disease was performed utilizing biotinylated Concanavalin-A (Con A), wheat germ agglutinin (WGA), and peanut agglutinin (PNA), in conjunction with an avidin-biotin peroxidase complex. Hydatidiform mole, invasive mole and choriocarcinoma exhibited increased receptors to Con-A and WGA compared to normal placenta. Increased reactivity to Con-A and WGA was associated merely with increased growth and proliferation of trophoblasts rather than a malignant transformation. Normal placenta, partial and complete mole generally showed moderate to strong binding with PNA after neuraminidase treatment, while invasive mole and choriocarcinoma (11 of 15 cases) generally showed minimal to absent reaction with PNA. Heterogeneity of PNA binding in choriocarcinoma was manifested by the presence of PNA reactivity in the trophoblast membrane in 2 cases wherein no prior neuraminidase treatment was given. This suggests that in some malignant trophoblasts, there is absence of sialic acid in the terminal cell surface carbohydrate groups resulting in the exposure of N-acetylgalactoseamine. PMID- 2560370 TI - [Catalytic properties of acetylcholinesterase, modified by N,N-dimethyl-2 phenylaziridinium. An alkane sulfonation reaction]. AB - By means of affinity labelling with N,N-dimethyl-2-phenylaziridinium ion (DPA) two forms of acetylcholinesterase were synthesized that contained one or two molecules of the label covalently attached to the enzyme. The reaction of native and covalently modified acetylcholinesterases with n-alkane sulfonyl chlorides CnH2n + 1SO2Cl at n = 1 -4 was used to characterize the reactivity and properties of the enzymes. It was found that labelling of acetylcholinesterase with one molecule of DPA did not affect the enzyme's reactivity. Acetylcholinesterase containing two labels (the second one presumably located at the anionic centre of the enzyme) displayed enhanced and more specific reactivity towards alkane sulfonyl chlorides. It was found that the phenomenon of acceleration caused by affinity modification is analogous to the influence of n-tetraalkylammonium ions on the same reaction. Therefore, the mechanism of regulation of the properties of the esteric centre, caused by affinity labelling of the enzyme at the anionic centre, is the same as in the case of n-tetralkylammonium ions. PMID- 2560371 TI - Synthesis and angiotension converting enzyme inhibitory activity of L-lysyl-N substituted glycine derivatives. AB - The synthesis and biological activity of novel L-lysyl-N-substituted glycine derivatives which exhibit in vitro and in vivo angiotensin converting enzyme (ACE) inhibition, are described. Particularly, N-[N alpha-(1-carboxy-3 phenylpropyl)-L-lysyl]-N-(4-phenylcyclohexyl)glycine (11e), N-[N alpha-(1-carboxy 3-cyclopentylpropyl)-N epsilon-carbobenzoxy-L- lysyl]-N-cyclopentyl glycine (9h) and N-[N alpha-[1(S)-carboxy-3-cyclohexylpropyl]-L-lysyl]-N-cyclopentyl glycine (19a) showed a strong inhibitory activity at IC50 as low as 0.65, 0.64 and 0.11 nmol/l, respectively. The most potent activity in vivo was observed with the compound 19a after i.v. administration in the rat. PMID- 2560372 TI - Reaction of pyridoxamine-5-P with pyrroloquinoline quinone (coenzyme PQQ). AB - PQQ catalyzes the oxidation of pyridoxamine (PM) and pyridoxamine-5-P (PMP) to pyridoxal and pyridoxal-5-P (PLP) at 37 degrees C in the absence of micelles and proteins. The time course of conversion of PMP into PLP was monitored by absorption spectroscopy; a rate of 10 nmol PLP/min was determined. The product of the reaction was identified by TLC, HPLC and its ability to restore the catalytic activity of apoaspartate aminotransferase. The conversion of PMP into PLP by free PQQ is more efficient than reactions catalyzed by the enzymes plasma amine oxidase and pyridoxamine-5-P oxidase at optimal pH values. PMID- 2560373 TI - PQQ and quinoprotein research--the first decade. AB - On the occasion of the first international symposium on pyrroloquinoline quinone (PQQ) and quinoproteins (Delft, September 1988), a review of this novel field in enzymology is presented. Quinoproteins (PQQ-containing enzymes) are widespread, from bacteria to mammalian organisms (including man), and occur in several classes of enzymes. Indications already exist that PQQ is a versatile cofactor, involved not only in oxidation but also in hydroxylation, transamination, decarboxylation and hydration reactions. The current list of quinoproteins shows that it was overlooked in several well-studied enzymes where the presence of a common cofactor had already been established. Up until now, all eukaryotic quinoproteins have covalently bound PQQ (or perhaps pro-PQQ), while free PQQ occurs exclusively in a number of (bacterial) dehydrogenases and in the culture fluid of certain Gram-negative bacteria. Biosynthesis of free PQQ in methylotrophic bacteria starts with tyrosine and glutamic acid as precursors while intermediates in the route have not been detected and the presence of free PQQ is not required for synthesis of the covalently bound form of the cofactor in glutamic acid decarboxylase from Escherichia coli. Therefore, the assembly of covalently bound cofactor might occur in situ, i.e. in the quinoproteins themselves. If the latter also applies to mammalian quinoproteins, this implies that PQQ is not a vitamin. On the other hand, positive effects have been reported upon administration of PQQ to test animals. Methods suited to detach and to detect PQQ with a derivatized o-quinone moiety may answer questions on the uptake and processing of the compound. PMID- 2560374 TI - Derivatives of glutamic acid and trehalose which can be transformed into long living free radicals by the loss of an electron are identified in some bacteria. AB - A derivative of glutamic acid (ammonigenin) and a trisaccharide named lysodektose which are converted into long-living free radicals by the loss of one electron were isolated from Brevibacterium ammoniagenes and Micrococcus lysodeikticus. Structural formulae suggested for both substances based on ESR-, NMR- and mass spectra, isotopic substitution experiments and other data are: lactone of N hydroxy-N-(2-carbamoylethyl)-glutamyl-4-amino-2-hydroxybutyric amide and 6-O-[2 deoxy-2-(N-methyl)-hydroxylamino-beta-D-glucopyranosyl]- alpha, alpha-trehalose. Radical forms appear on reversible oxidation of hydroxylamino groups to nitroxyl groups. Participation in the protection of bacterial cells and regulation of their metabolism is suggested for these compounds. PMID- 2560375 TI - [Domestic and occupational pneumo-allergens among workers with tropical wood in Abidjan (Ivory Coast)]. AB - The authors realized an epidemiologic research by skin test method in a furniture factory in Abidjan (Ivory Coast). 19 wood workers underwent these tests: their professional risk to wood dust exposure was over 13 years. The results show the importance of allergenic work atmosphere and mainly the high allergic potential of five exotic woods currently used. This article proposes an approach of the specific immunological pathways through a prospective allergological enquiry conducted at furniture factory in Abidjan (Ivory Coast). PMID- 2560376 TI - Specific inhibitory substance to the husband's HLA allotype in sera from patients with gestational trophoblastic disease detected by lymphocytotoxicity testing. AB - The sera of nine patients with gestational trophoblastic disease were investigated for the inhibitory substance specific to the HLA allotypes of the patients' husbands by microcytotoxicity inhibition testing using monoclonal antibodies (anti-A2, A9, A10) and HLA-typing serum to Bw52. The specific inhibitory substance was present in three of three choriocarcinoma patients, in three of four invasive mole patients, and in one of two hydatidiform mole patients and seemed to change in titer parallel with tumor growth. These facts suggest that the inhibitory substance could have some relation to the patient's immunological response against the tumor; that is, this inhibitory substance could be the reason why patients could not induce graft rejection response to trophoblastic tumor cells. PMID- 2560378 TI - Effect of sodium bicarbonate and sodium starch glycolate on the in vivo disintegration of hard gelatin capsules--a radiological study in the dog. AB - The release of drugs from hard gelatin capsules is often limited by the disintegration rate of the capsule. We set out to determine whether it is possible to hasten the in vivo disintegration of hard gelatin capsules by adding disintegrants (sodium bicarbonate or sodium starch glycolate) to the formulation. This radiological study was carried out in six beagle dogs. We conclude that if rapid disintegration is desired for a hard gelatin capsule formulation, water soluble diluents should primarily be selected. Sparingly water-soluble, gelforming diluents cause slow in vivo disintegration. Using swellable disintegrants such as sodium starch glycolate does not accelerate disintegration and may even retard it. With sodium bicarbonate it is possible to shorten the disintegration time of capsules containing water-insoluble ingredients, but not to the extent possible with water-soluble diluents. PMID- 2560377 TI - Prostacyclin release and cytotoxicity of peritoneal cells are inversely related in pregnant and non-pregnant mice infected with herpes simplex virus. AB - Cytotoxicity of peritoneal cells in a HSV-infected murine model is attenuated in late pregnancy. Prostacyclin (PGI2) is elevated at this time in reproductive tissues and has been implicated in the regulation of the immune response. The purpose of this study was to estimate PGI2 in the peritoneal wash or culture supernatants of peritoneal cells obtained from uninfected and HSV-infected pregnant and virgin mice using a radioimmunoassay for 6-keto-prostaglandin F1 alpha. The peritoneal wash of uninfected pregnant and virgin mice contained high levels of 6-keto-PGF1 alpha, 505 +/- 51 pg/100 microliters, (mean +/- S.E., n = 15), and 200 +/- 19 pg/100 microliters, (n = 30), ad did peritoneal effector and target cell cultures (1,159 +/- 118 pg/100 microliters, n = 6, and 1,057 +/- 207 pg/100 microliters, n = 7), respectively. HSV-infection induced in vitro cytotoxicity and suppressed the release of 6-keto-PGF 1 alpha (r = -0.897, P less than 0.05, n = 18). Its concentration was significantly higher (14-fold, P less than .05) in the peritoneal wash, but not in the cell culture, of pregnant (212 +/- 29 pg/100 microliters, n = 19) as compared to virgin mice (18.5 +/- 3.4 pg/100 microliters, n = 27). The levels of 6-keto-PGF1 alpha were inversely correlated (P less than .05) with the combined effects of HSV-infection and cytotoxicity. PMID- 2560379 TI - Antigen processing and presentation. PMID- 2560380 TI - Lymphocyte activation. PMID- 2560381 TI - Cytokines: structure, function and synthesis. PMID- 2560382 TI - Metastatic breast carcinoma of the orbit. AB - Two Caucasian females in whom an orbital mass was the first sign of metastasis from a breast carcinoma are described. Each subject had undergone a radical mastectomy to treat a primary breast carcinoma, one six years and the other eight years previously. Each presented with ipsilateral proptosis, and restricted upper eyelid action; one had altered ocular motility. In each subject an open biopsy of the orbital mass via an eyelid crease incision revealed metastatic scirrhous adenocarcinoma of the breast. The clinical features, investigations, and treatment of orbital metastases from breast carcinoma are discussed. PMID- 2560383 TI - Acute non-haemorrhagic conjunctivitis due to coxsackievirus A24. AB - Two cases of acute non-haemorrhagic conjunctivitis due to coxsackievirus A24 (CA24) are described. These are the first Australian isolates. The presentation was as a severe conjunctivitis in otherwise healthy adults who had not travelled outside Australia. The course was of short duration and self-limiting with no long-term sequelae. The isolates could not be neutralised by antiserum prepared against prototype CA24 but were identified by immune electron microscopy and complement fixation. PMID- 2560384 TI - High-dose intravitreal gancyclovir for cytomegaloviral (CMV) retinitis. AB - A patient with acquired immune deficiency syndrome (AIDS) with bilateral cytomegaloviral (CMV) retinitis who had become neutropaenic from intravenous gancyclovir, was treated with a total of 16 intravitreal 2 mg/0.1 ml doses of gancyclovir in each eye under topical anaesthesia. The injections were well tolerated with no evidence of retinal toxicity. A moderate vitreous haze which did not affect vision was the only complication. The patient retained useful vision until the time of his death, three months later. PMID- 2560385 TI - Structure and developmental expression of the nerve growth factor receptor in the chicken central nervous system. AB - Chicken nerve growth factor (NGF) receptor cDNAs have been isolated and sequenced in an effort to identify functionally important receptor domains and as an initial step in determining the functions of the NGF receptor in early embryogenesis. Comparisons of the primary amino acid sequences of the avian and mammalian NGF receptors have identified several discrete domains that differ in their degree of conservation. The highly conserved regions include an extracellular domain, likely to be involved in ligand binding, in which the positions of 24 cysteine residues and virtually all negatively charged residues are conserved; a transmembrane region, including flanking stretches of extracellular and cytoplasmic amino acids, which has properties suggesting it interacts with other proteins; and a cytoplasmic PEST sequence, which may regulate receptor turnover. Transient expression of NGF receptor mRNA has been seen in many regions of the developing CNS. Experiments suggest that both NGF and its receptor help regulate development of the retina. PMID- 2560386 TI - Channel expression correlates with differentiation stage during the development of oligodendrocytes from their precursor cells in culture. AB - Membrane currents in cultured murine oligodendrocytes and their precursors were characterized using the patch-clamp technique. Prior to recording, cells were identified by immunofluorescence using monoclonal antibodies characteristic of two types of precursor cells and two differentiation stages of oligodendrocytes. The most immature, A2B5 antigen-positive glial precursors, expressed four types of voltage-activated K+ currents and tetrodotoxin-sensitive Na+ currents. The more differentiated cells, O4 antigen-positive glial precursors, expressed similar K+ currents, but Na+ currents were recorded in only a minority of cells. In differentiated O1 and O10 antigen-positive oligodendrocytes the channels characteristic of precursor cells were no longer observed, but an inwardly rectifying K+ current was apparent. Thus, channel expression by cells of the oligodendrocyte lineage correlates with differentiation stage and is more complex in precursor cells than in oligodendrocytes. PMID- 2560387 TI - Differential G protein-mediated coupling of neurotransmitter receptors to Ca2+ channels in rat dorsal root ganglion neurons in vitro. AB - The peptides neuropeptide Y (NPY) and bradykinin (BK) both inhibited Ca2+ currents in rat dorsal root ganglion neurons (DRG) in vitro. The effects of both peptides were completely blocked by treatment of cells with pertussis toxin. Based on antigenic determinants, DRG cells contained at least two pertussis toxin substrates, alpha o (Mr, 39 kd) and alpha i2 (Mr, 40 kd). We examined the ability of three purified bovine alpha subunits (identified with antibodies as alpha o, alpha i1, and alpha i2) to reconstitute the inhibitory effects of NPY and BK. Reconstitution of NPY effects occurred according to the potency series alpha o greater than alpha i1 much greater than alpha i2. However, in the case of BK all three G proteins were approximately equally effective. Whereas complete reconstitution of NPY effects could be obtained with alpha o, no single alpha subunit produced complete reconstitution of BK. Combinations of alpha o and alpha i2, however, were able to completely reconstitute the effects of BK. Thus several G proteins can effect the regulation of Ca2+ channels in these cells. However, neurotransmitters may be selective in the G proteins or combinations of G proteins utilized to achieve this regulation. PMID- 2560388 TI - Signal sorting and amplification through G protein-coupled receptors. PMID- 2560390 TI - Distribution of Na+ channels and ankyrin in neuromuscular junctions is complementary to that of acetylcholine receptors and the 43 kd protein. AB - We have used immunogold electron microscopy to study the organization of the acetylcholine receptor, 43 kd protein, voltage-sensitive Na+ channel, and ankyrin in the postsynaptic membrane of the rat neuromuscular junction. The acetylcholine receptor and the 43 kd protein are concentrated at the crests of the postsynaptic folds, coextensive with the subsynaptic density. In contrast, Na+ channels and ankyrin are concentrated in the membranes of the troughs and in perijunctional membranes, both characterized by discontinuous submembrane electron-dense plaques. This configuration of interspersed postsynaptic membrane domains enriched in either Na+ channels or acetylcholine receptors may facilitate the initiation of the muscle action potential. Furthermore, the results support the involvement of ankyrin in immobilizing Na+ channels in specific membrane domains, analogous to the proposed involvement of the 43 kd protein in acetylcholine receptor immobilization. PMID- 2560389 TI - On the resting potential of isolated frog sympathetic neurons. AB - One of the oldest questions of electrophysiology, the origin of the resting potential, has yet to be answered satisfactorily for most cells. Isolated frog sympathetic neurons, studied with whole-cell recording, generally have resting potentials of approximately -75 mV with an input resistance of approximately 300 M omega. These properties are not expected from the M-type K+ current (IM) or from other ionic currents previously described in these cells. In the -60 to -110 M mV voltage region, at least three currents are present: an inwardly rectifying current (IQ), a resting current with little voltage sensitivity carried at least in part by K+, and a (Na+,K+)ATPase pump current. The resting K+ current, not IM or IQ is the primary ionic current near the resting potential under these conditions. The electrogenic pump contributes an additional approximately 10 mV of hyperpolarization. PMID- 2560391 TI - Antibodies to the GTP binding protein, Go, antagonize noradrenaline-induced calcium current inhibition in NG108-15 hybrid cells. AB - The voltage-dependent calcium current in chemically differentiated NG108-15 cells is depressed by noradrenaline acting on alpha-adrenoreceptors. The response is absent in cells pretreated with pertussis toxin, implicating the involvement of a G-protein. To identify this G-protein, we have studied the response to noradrenaline in cells preinjected with antibodies specific for two G-proteins, Gi and Go. Cells injected with the Gi antibody responded normally to noradrenaline. In contrast, the response to noradrenaline in cells injected with the Go antibody was markedly attenuated. We conclude that Go is employed in coupling alpha-adrenoreceptors to the calcium channels in NG108-15 cells. PMID- 2560392 TI - Seizure-like activity and cellular damage in rat hippocampal neurons in cell culture. AB - Neurons dissociated from the hippocampal formations of neonatal rats were grown in medium containing kynurenic acid (a glutamate receptor antagonist) and elevated Mg2+. Such chronically blocked neurons, when first exposed to medium without blockers (after 0.5-5.0 months), generated intense seizure-like activity. This consisted of bursts of synchronous electrical responses that resembled paroxysmal depolarization shifts and sustained depolarizations that, in some neurons, nearly abolished the resting potential. Sustained depolarizations were usually reversed by timely application of kynurenate or 2-amino-5 phosphonovalerate, indicating that continuous activation of glutamate receptors was required for their maintenance. Prolonged periods of intense seizure-like activity usually killed most neurons in the culture. This system allows seizure related cellular mechanisms to be studied in long-term cell culture. PMID- 2560393 TI - NGF induction of NGF receptor gene expression and cholinergic neuronal hypertrophy within the basal forebrain of the adult rat. AB - Chronic infusion of nerve growth factor (NGF) into the forebrain of the adult rat produced increases in NGF receptor (NGF-R) mRNA hybridization, NGF-R immunoreactivity, choline acetyltransferase (ChAT) mRNA hybridization, and neuronal hypertrophy, when compared with vehicle infusion or noninfused rat brain. In situ hybridization showed NGF induction of NGF-R gene expression, documented by increases in the number of NGF-R mRNA-positive cells within the medial septum, diagonal band, and nucleus basalis magnocellularis. NGF also produced hypertrophy of ChAT mRNA-positive neurons. These results suggest that NGF produces cholinergic neuronal hypertrophy through induction of NGF-R gene expression within the basal forebrain. PMID- 2560394 TI - Fetal lung fibroblasts secrete and respond to insulin-like growth factors. PMID- 2560395 TI - IgG specifically enhances chrysotile asbestos-stimulated superoxide anion production by the alveolar macrophage. AB - The interaction of chrysotile asbestos with alveolar macrophages in vitro is known to stimulate cellular superoxide anion production. However, it is likely that particulates in the respiratory tract are present together with components of the pulmonary surfactant, and it is not known how these components may alter the bioactivity of the particulate. We now show that guinea pig immunoglobulin G, a surfactant protein, causes a significant, dose-dependent enhancement of superoxide anion production by nonadherent guinea pig alveolar macrophages in response to chrysotile asbestos. This enhancement could not be mimicked by other particulates or proteins, including IgG fragments, implying that the interaction between IgG, cell, and chrysotile is relatively specific. The enhancing effect of IgG in solution could be reproduced by pretreating the chrysotile asbestos with IgG. The fact that IgG specifically enhances chrysotile asbestos-stimulated superoxide anion production, in turn, leads to a proposal for a molecular mechanism by which asbestos may stimulate the guinea pig macrophage, namely, by crosslinking cell-surface immunoglobulin Fc receptors. In view of the submicromolar concentrations at which IgG was effective in enhancing macrophage stimulation by chrysotile asbestos in vitro, these results also suggest that IgG adsorption may play a role in the progression of asbestos-induced pulmonary fibrosis. PMID- 2560396 TI - Molecular biology of avian infectious bronchitis virus. PMID- 2560397 TI - Egg drop syndrome. PMID- 2560398 TI - Breast lesions in generalized neurofibromatosis: breast cancer and cystosarcoma phylloides. AB - This report describes three cases from one surgical unit of the Mansoura Teaching Hospital of Cairo, Egypt, with generalized neurofibromatosis and associated breast lesions. Two patients had carcinoma of the breast and the third had cystosarcoma phylloides. PMID- 2560400 TI - Aneurysmectomy prognosticators by equilibrium multi-gated cardiac blood pool scintigraphy. AB - Non-invasive assessment of left ventricular aneurysm (LVA) resection prognosticators was obtained in 13 multigated cardiac blood pool scintigrams by applying Fourier analysis to identify regions of interest. Results were compared to contrast ventriculograms performed at recent cardiac catheterization. Excellent correlation of the contractile section ejection fraction (CSEF) was found between the two techniques, r = .92 (P less than or equal to .0001). The CSEF correlated inversely with the left ventricular end diastolic pressure (LVEDP), r = .83 (P less than or equal to .0004), implying that LVEDP is not an independent prognosticator. The relative LVA size expressed as a fraction of the left ventricle in end diastole (LVASF) correlated less strongly with LVASF derived from contrast ventriculography, r = .67 (P = .012). This was probably due to additive inherent errors in contrast ventriculography geometric assumptions. Paradoxical stroke steal fraction (PSSF) defined as the reverse stroke lost in the LVA divided by the left ventricular forward stroke, was calculated from subtraction images. We propose a resectability score utilizing the product of the scintigraphically derived CSEF X LVASF X PSSF X 1,000 to obtain non-invasive pre operative prognostication of LVA resection and post-operative evaluation. PMID- 2560401 TI - Types of human papillomavirus isolated from Japanese patients with epidermodysplasia verruciformis. AB - Virological studies were performed on 12 patients with epidermodysplasia verruciformis (EV). Three types of lesions were observed: red plaques, pityriasis versicolor (PV)-like macules and plane warts. Human papillomavirus (HPV) 14, 20 and 21 were isolated from the plaques, HPV 3, 14 and 38 from flat warts and HPV 5, 12, 17, 20 and 38 from PV-like lesions. No clear relationship could be established between the different lesions and the types of HPV. Types 17 and 20 have been isolated most frequently from Japanese EV patients and HPV 5, frequently detected in other countries, is less common, whereas HPV 8 has not been isolated. Skin cancers occurred in six of the cases (50%) and all had benign lesions that were PV-like. At least one type of HPV 5, 17 or 20 could be isolated from these benign lesions and HPV 17 or 20 detected in the cancers. These three types of HPV in EV patients appear to be involved in the malignant transformation. PMID- 2560399 TI - Recombinant human mullerian inhibiting substance inhibits epidermal growth factor receptor tyrosine kinase. AB - Autophosphorylation of the epidermal growth factor (EGF) receptor in A-431 cells and plasma membrane fractions was inhibited by partially purified recombinant human Mullerian Inhibiting Substance (MIS). Immunoprecipitation of the EFG receptor using anti-EGF receptor or anti-phosphotyrosine antibodies, and phosphoamino acid analysis of this receptor, demonstrated that MIS specifically inhibited EGF-induced tyrosine phosphorylation. Inhibition of EGF receptor autophosphorylation by MIS in membrane preparations was not affected by increasing concentrations of EGF, manganese or [gamma-(32)P] ATP. Thus, it is unlikely that MIS competes for EGF binding sites or sequesters substrate. Immunoabsorption of MIS with anti-human MIS antibody blocked the MIS inhibition of EGF receptor autophosphorylation, indicating that the inhibition was due to MIS. Our data suggest that MIS regulates the activity of the EGF receptor tyrosine kinase in A-431 cells. PMID- 2560402 TI - Transmission of virus particles by cryotherapy and multi-use caustic pencils: a problem to dermatologists? AB - The possibility of transmission of virus particles from patient to patient by multi-use silver nitrate sticks and styptic pencils and by cotton wool swabs that have been dipped repeatedly into Dewar flasks of liquid nitrogen was studied. The results showed that transmission by the sticks and pencils is unlikely but that virus particles can be transferred into and out of a Dewar flask on cotton wool swabs. We suggest that swabs should not be dipped repeatedly into the flask of liquid nitrogen but that, instead, a small aliquot of nitrogen should be decanted into a smaller 'clean' vessel and a new cotton swab used for each patient. PMID- 2560403 TI - Variegate porphyria associated with hepatocellular carcinoma. AB - A case is described of hepatocellular carcinoma presenting with the cutaneous and biochemical features of variegate porphyria. PMID- 2560404 TI - Effect of placental factors on growth and function of the human fetal adrenal in vitro. AB - Conditioned medium from human placental monolayer cultures (PM) had a marked stimulatory effect on proliferation (3H-thymidine uptake) of human fetal zone adrenal cells in primary monolayer culture, even in the absence of serum. Epidermal growth factor (EGF) and fibroblast growth factor (FGF) also significantly stimulated fetal adrenal cell growth. However, the effects of PM differed from those of EGF and FGF in several respects: 1) maximal response to PM was 2-5 times greater; 2) mitogenic effects of EGF and FGF were suppressed by adrenocorticotropic hormone (ACTH), whereas that of 50% PM was not; 3) PM inhibited ACTH-stimulated steroidogenesis (dehydroepiandrosterone sulfate and cortisol), but EGF and FGF did not. Preliminary characterization studies have indicated that approximately half of the placental growth-promoting activity is heat resistant and sensitive to bacterial proteases, and that 50-60% of the activity is lost after dialysis with membranes having a molecular weight cutoff of 3500. These findings suggest a role for the placenta in the growth and differentiated function of the human fetal adrenal gland. PMID- 2560405 TI - The response of large and small luteal cells from the pregnant rat to substrates and secretagogues. AB - Large (greater than 22 microns) and small (12-21 microns) luteal cells from Day 8 pregnant rats were separated by elutriation after enzyme dissociation. Aliquots of cells were incubated for 4 h at 37 degrees C in Medium 199 alone (control) or with medium containing dibutyryl cyclic adenosine 3', 5'-monophosphate (cAMP) at 0.5 mM or 5 mM; rat luteinizing hormone (LH) at doses of 1, 10, 100, or 1000 ng/ml; 10 micrograms/ml 25-OH-cholesterol; or 10 ng/ml testosterone. Production of progesterone, testosterone, and estradiol was measured by radioimmunoassay. Both cell types showed a similar increase in estradiol synthesis when stimulated with LH (1 microgram/ml) or dibutyryl cAMP (5 mM); however, large luteal cells aromatized exogenous testosterone, whereas small luteal cells did not. Large luteal cells produced increased amounts of progesterone at lower doses of dibutyryl cAMP (0.5 mM) and LH (10 ng/ml), compared to small cells, which required 5 mM dibutyryl cAMP or 1 microgram/ml LH for minimal stimulation. Dibutyryl cAMP (5 mM) also resulted in an increase of testosterone release from small luteal cells. Progesterone synthesis in both cell types was enhanced by 25 OH-cholesterol. These results suggest that the two cell types differ functionally with respect to steroidogenesis during pregnancy, and that the large luteal cells appear to be the primary site of progesterone and estradiol production at this stage of pregnancy. PMID- 2560406 TI - The permeability of hydrophobic membranes to 22Na salts and 14CO2 in low dielectric media. AB - The one-way fluxes of 14CO2 and a series of 22Na (Cl, Br, HCO3, ClO4, I) salts across n-hexadecane-impregnated solid-support liquid membranes have been measured in water and low dielectric media (50-90 vol% dioxane/water). One-way fluxes for 14CO2 (J14CO2) were 0.84 and 1.03 x 10(-9) mol cm-2 s-1 in 75% dioxane (aq.) and water, respectively, across both impregnated cellulose and teflon membranes. 22Na fluxes across impregnated cellulose membranes in 75% dioxane (aq.) ranged from 1.8 to 11.4 x 10(-10) mol cm-2 s-1 and had the order NaCl less than NaBr less than NaHCO3 less than NaClO4 less than Nal. 22Na fluxes across impregnated teflon membranes were slightly smaller, 1.5-7.1 x 10(-10) mol cm-2 s-1, but had the same order for the anions tested. No measurable 22Na fluxes were observed in aqueous media. For NaI and NaClO4 there was a 3-6-fold enhancement of fluxes in 90% dioxane (aq.) compared to 75% dioxane (aq.). The corresponding enhancement for fluxes of NaHCO3, NaBr and NaCl was 1.5-fold. The results are discussed in terms of ion-paired salt transport in low dielectric media. PMID- 2560407 TI - Growth factors and oncogenes. AB - Recent findings reporting the structure, the mechanism of action of peptide growth factors and their receptor as well as their presence in human tumors have led to a better knowledge of the events involved in cell growth regulation. The ability of cancer cells to produce and to respond to their own growth factors confers on them a growth autonomy and strengthens the link between growth factors and oncogenes. Oncogenes render the cells growth factor-independent not only by coding for autocrine growth factors of their receptor, but also by amplifying the mitogenic signals generated by the binding of the peptide growth factor to its membrane receptor. PMID- 2560408 TI - Treatment of migraine with salmon calcitonin: effects on plasma beta-endorphin, ACTH and cortisol levels. AB - In this study we have examined the results of salmon calcitonin treatment on migraine pain. The mechanism by which calcitonin induces analgesia is still not understood. We observed the effect of a 5-day treatment with salmon calcitonin (IM 100 IU/day) on circulating levels of beta-endorphin, ACTH, and cortisol in 20 patients with migraine during the headache-free period. All 3 hormones were increased after the calcitonin administration and the maximum increase was obtained in beta-endorphin levels. There were significant statistical correlations between beta-endorphin, ACTH, and cortisol levels determined before and after calcitonin treatment. PMID- 2560409 TI - Biology of gut anaerobic fungi. AB - The obligately anaerobic nature of the gut indigenous fungi distinguishes them from other fungi. They are distributed widely in large herbivores, both in the foregut of ruminant-like animals and in the hindgut of hindgut fermenters. Comparative studies indicate that a capacious organ of fermentative digestion is required for their development. These fungi have been assigned to the Neocallimasticaceae, within the chytridiomycete order Spizellomycetales. The anaerobic fungi of domestic ruminants have been studied most extensively. Plant material entering the rumen is rapidly colonized by zoospores that attach and develop into thalli. The anaerobic rumen fungi have been shown to produce active cellulases and xylanases and specifically colonise and grow on plant vascular tissues. Large populations of anaerobic fungi colonise plant fragment in the rumens of cattle and sheep on high-fibre diets. The fungi actively ferment cellulose which results in formation of a mixture of products including acetate, lactate, ethanol, formate, succinate, CO2 and H2. The properties of the anaerobic fungi together with the extent of their populations on plant fragments in animals on high-fibre diets indicates a significant role for the fungi in fibre digestion. PMID- 2560410 TI - Are glial cells targets of the central noradrenergic system? A review of the evidence. AB - It has been suggested by a number of investigators that glial cells as well as neurons are targets of the central noradrenergic system. This important hypothesis, however, has not been presented previously in a systematic and unified manner. The present review was therefore undertaken to accomplish this. The evidence supporting noradrenergic action on glia consists primarily of findings that beta-adrenoceptors, norepinephrine (NE)-stimulated cyclic AMP (cAMP) responses and glycogen are localized preferentially in glial cells and that beta-receptor density and glycogen hydrolysis are under the control of neuronally released NE. While there is some disagreement as to the extent to which beta-receptors are preferentially localized in glia, there is a consensus that most glycogen in the forebrain is localized in this cellular compartment. The presumed function of the noradrenergic action on glia appears to be the release of glucose for production of energy, the synthesis of neurotrophic factors such as nerve growth factor, and the release of substances which may affect local neurotransmission including taurine, cAMP and its metabolites. These glial responses may be intimately related to the electrophysiological actions of NE on neurons. PMID- 2560411 TI - [Serological investigation on infant virus pneumonia in Beijing]. AB - Antibody against adenovirus in paired sera of 171 cases of infant virus pneumonia observed in 7 winter-spring periods was assayed by hemagglutination-inhibition test and enzyme-linked immunosorbent assay. A 4-fold or greater rise in the titer was found in 56 cases (32.75%). Type 3 and Type 7 were predominant. Antibody against influenza virus was assayed in paired sera of 129 cases in 6 winter spring periods. A 4-fold or greater increase in the titer was observed in 11 cases (8.53%), with predominance of A1 and A3. Antibody against parainfluenza virus was also assayed in paired sera of these 129 cases. A 4-fold or more rise in the titer was demonstrated in 27 cases (20.93%), with predominance of Type 3. Antibody against respiratory syncytial virus was assayed in paired sera of 60 cases. A 4-fold or greater increase in the titer was shown in 14 cases (23.30%). PMID- 2560412 TI - [Polyacrylamide gel electrophoresis (PAGE) as a method for detecting enteric adenovirus (EAd)]. AB - This paper presents a method which could provide a simple, rapid, economical, and reliable means of detecting or identifying adenoviruses (Advs), rotaviruses (RVs) and reoviruses (ReoVs) in stool suspensions or tissue cultures. The method is based on polyacrylamide gel electrophoresis (PAGE) of the virus nucleic acids, but sample preparation does not need the use of radioactive label, specific DNA probe or antisera. Comparison of the results of PAGE of Adv, RV and ReoV with those of electron microscopy (EM) and/or enzyme-linked immunoabsorbent assay (ELISA) was made. The method is of comparable sensitivity to electron microscopy, and is not limited by amount of specimens obtained, and is thus suitable for application as a batch testing method. PMID- 2560413 TI - Immunofluorescence evidence for cytoskeletal rearrangement accompanying pigment redistribution in goldfish xanthophores. AB - Immunofluorescence and phase-contrast microscopic studies of goldfish xanthophores with aggregated or dispersed pigment show two unusual features. First, immunofluorescence studies with anti-actin show punctate structures instead of filaments. These punctate structures are unique for the xanthophores and are absent from both goldfish dermal non-pigment cells and a dedifferentiated cell line (GEM-81) derived from a goldfish xanthophore tumor. Comparison of immunofluorescence and phase-contrast microscopic images with electron microscopic images of thin sections and of Triton-insoluble cytoskeletons show that these punctate structures represent pterinosomes with radiating F-actin. The high local concentration of actin around the pterinosomes results in strong localized fluorescence such that, when the images have proper brightness for these structures, individual actin filaments elsewhere in the cell are too weak in their fluorescence to be visible in the micrographs. Second, whereas immunofluorescence images with anti-tubulin show typical patterns in xanthophores with either aggregated or dispersed pigment, namely, filaments radiating out from the microtubule organizing center, immunofluorescence images with anti-actin or with anti-intermediate filament proteins show different patterns in xanthophores with aggregated versus dispersed pigment. In cells with dispersed pigment, the punctate structures seen with anti-actin are relatively evenly distributed in the cytoplasm, and intermediate filaments appear usually as a dense perinuclear band and long filaments elsewhere in the cytoplasm. In cells with aggregated pigment, both intermediate filaments and pterinosomes with associated actin are largely excluded from the space occupied by the pigment aggregate, and the band of intermediate filaments surrounds not only the nucleus but also the pigment aggregate. The patterns of distribution of the different cytoskeleton components, together with previous results from this laboratory, indicate that formation of the pigment aggregate depends at least in part on the interaction between pigment organelles and microtubules. The possibility that intermediate filaments may play a role in the formation/stabilization of the pigment aggregate is discussed. PMID- 2560414 TI - Preparation of microtubules from rat liver and testis: cytoplasmic dynein is a major microtubule associated protein. AB - A microtubule associated protein from brain tissue (MAP 1C), has been found to possess many properties in common with ciliary and flagellar dyneins (Paschal et al.:J. Cell Biol. 105:1273-1282, 1987). However, this protein, now designated as cytoplasmic dynein, exhibited several properties which distinguish it from axonemal forms of the enzyme. We have investigated these characteristics further in a study of cytoplasmic dyneins from non-neuronal tissues. Rat liver and testis in particular were found to contain high levels of cytoplasmic dynein. The yield of dynein from testis was over 70 micrograms/g of tissue, making this the best source of cytoplasmic dynein of all tissues so far examined. The characterization of dynein from these sources has confirmed and extended our previous observations concerning the unique properties of cytoplasmic dynein. Activation of liver and testis dynein occurred at low (less than 1 mg/ml) tubulin concentration. Polypeptides identified as subunits of brain cytoplasmic dynein (74, 59, 57, 55, and 53 kDa) were present in liver and testis preparations. In addition, polypeptides at 150 and 45 kDa were found to copurify with the non-neuronal dyneins. The liver and testis enzyme hydrolyzed pyrimidine nucleotides at rates up to 12.5 times faster than ATP, though the relative affinity of cytoplasmic dynein for CTP was much lower (Km = 1.0 mM) than that for ATP. The properties of the testis enzyme were consistent with its identification as a cytoplasmic dynein rather than a sperm axonemal precursor. These data indicate that cytoplasmic dyneins may be widespread in distribution and that they share certain biochemical properties unique from those of axonemal dyneins. These characteristics are consistent with the proposal that cytoplasmic dynein plays a universal role in retrograde organelle motility. PMID- 2560415 TI - [Askin tumor. Rare neurogenic tumor with metastasis to mouth, jaw and face regions]. AB - One case of a malignant peripheral neuroectodermal tumor of the thoracopulmonary region (Askin-Tumor) is reported. The tumor represents a distinct pathologic entity of neuroectodermal origin. The patient developed a metastasis in the upper jaw and in a lumbar vertebra. Morphologic and immunocytochemical findings of this extremely rare tumor were described and differential diagnosis from other soft tissue round-cell sarcomas, such as Ewing's sarcoma and neuroblastoma are discussed. Problems associated with the clinical behavior and treatment are outlined. The prognosis in most cases of Askin-tumor is poor. The recommended therapy is radical resection, irradiation and combination chemotherapy containing anthracyclines and a high dose of alkylating agents. PMID- 2560416 TI - [Antineuronal antibody in carcinomatous neuropathy]. AB - A seventy-year-old woman was admitted to the hospital because of paresthesias and progressive weakness of the extremities. Ten months previously, tightness in the thighs and numbness in the fingers developed, with subsequent tingling sensations of the hands and feet. Three months before entry, she had difficulty in walking because of severe paresthesias in the extremities. She was found to have an abnormal mass in a X-ray film of the chest, and transferred to our hospital. Neurological examination showed generalized hyporeflexia, marked loss of deep and discriminative sensations in the distal portion of the limbs, positive Romberg sign and severely ataxic gait. Cerebellar sign was negative. Cytologic examination of a transbronchial lung biopsy disclosed small cell carcinoma of the lung. A sural nerve biopsy showed severe loss of myelinated fibers accompanied by axonal degeneration of residual fibers. Immunohistochemically antineuronal antibody was found in the serum but not in the CSF, which reacted to nuclei and cytoplasm of the dorsal root ganglion neurons as well as Purkinje cells. Titers of this autoantibody in the serum were markedly reduced after chemotherapy and irradiation. Western blot analysis demonstrated that autoantibody in the serum recognized neuronal antigen with single band at 38 kd. Pathogenesis of carcinomatous neuropathy is still unexplained, but the findings here presented have given rise to the speculation that antineuronal antibody may be a contributory factor in carcinomatous neuropathy. PMID- 2560417 TI - The insulin receptor. PMID- 2560418 TI - Replacement of special enzyme immunoassay transport medium by a standard viral transport medium in the Herpcheck herpes simplex virus antigen detection test. AB - A new direct herpes simplex virus antigen enzyme immunoassay (EIA) uses a special EIA transport medium (ETM) for transport of herpes simplex virus (HSV) specimens. As ETM lyses the virus precluding culture and typing, we evaluated the relative performance of this EIA when performed on specimens transported in either ETM or a standard viral transport medium (VTM). These EIA results were also compared to cell culture performed on specimens transported in VTM (VTM-CC) and direct rapid inoculation into cell culture (CC). Based on all confirmed positives, by any test, the sensitivities for CC was 97% (66/68), for VTM-CC 91% (62/68), for ETM EIA 97% (66/68), and for VTM-EIA 93% (63/68). It appears that VTM may be a slightly less desirable substitute than ETM in the performance of EIA. However, VTM-EIA is certainly as sensitive as cell culture performed on VTM. PMID- 2560419 TI - Tazobactam prevention of emergence of resistance. PMID- 2560420 TI - Comparative in vitro activity of piperacillin combined with the beta-lactamase inhibitor tazobactam (YTR 830). AB - Combination with tazobactam substantially enhanced the activity of piperacillin against routine isolates of staphylococci, various Enterobacteriaceae, Acinetobacter anitratus, and Bacteroides fragilis. Tazobactam enhanced the activity of piperacillin more than fourfold against Pseudomonas aeruginosa spp. harboring eight of 12 plasmid-mediated beta-lactamases. PMID- 2560421 TI - Piperacillin/tazobactam (YTR 830) combination. Comparative antimicrobial activity against 5889 recent aerobic clinical isolates and 60 Bacteroides fragilis group strains. AB - Piperacillin combined with tazobactam (formerly YTR 830) was tested at a ratio of 8:1 against 5889 aerobic isolates and 50 strains from the Bacteroides fragilis group. Imipenem was the most active agent tested against Enterobacteriaceae (99.3% at less than or equal to 4 micrograms/ml), ceftazidime was most effective against nonenteric Gram-negative bacilli (80.7% at less than or equal to 8 micrograms/ml), and piperacillin/tazobactam possessed a superior spectrum against Gram-positive cocci (92.2% at less than or equal to 16/2 micrograms/ml). Against all aerobic strains, piperacillin/tazobactam had a spectrum (90.3% at less than or equal to 16/2 micrograms/ml) comparable to imipenem (93.6% at less than or equal to 4 micrograms/ml) and was distinctly greater than that of ticarcillin/clavulanic acid (73.3% at less than or equal to 16/2 micrograms/ml) and ceftazidime (75.5% at less than or equal to 8 micrograms/ml). Against the B. fragilis group isolates, all piperacillin/tazobactam MICs were less than or equal to 64/8 micrograms/ml. This activity was superior to piperacillin alone (MIC:50, 8-64 micrograms/ml) and cefoxitin (MIC50, 4-64 micrograms/ml). Piperacillin/tazobactam appears to be a promising parenteral antimicrobial combination, with a spectrum effective against a wide variety of clinical pathogens. PMID- 2560422 TI - Studies to optimize the in vitro testing of piperacillin combined with tazobactam (YTR 830). AB - The combination of piperacillin and the beta-lactamase inhibitor tazobactam (formerly YTR 830) was studied to determine optimal disk concentrations and dilution testing conditions. In addition, the potency of the combination was compared to that of piperacillin alone. The spectrum of piperacillin was greatly expanded by the addition to tazobactam principally against beta-lactamase producing strains of Haemophilus influenzae, Escherichia coli, Morganella morganii, Proteus vulgaris, Providencia stuartii, Shigella spp., Neisseria gonorrhoeae, and Staphylococcus spp. Tazobactam was active alone against Branhamella catarrhalis (minimum inhibitory concentration [MIC] 50, less than or equal to 1 microgram/ml), gonococci (MIC 50, 0.5-4 micrograms/ml), and N. meningitidis (MIC 50, less than or equal to 1 microgram/ml). Studies with beta lactamase-producing type strains showed tazobactam to have high affinity for plasmid-mediated enzymes (TEM-1 and 2, SHV-1, HMS-1, and some CARB or OXA types) and not chromosomal beta-lactamases. Piperacillin/tazobactam inhibited 93% of fluoro-quinolone resistant strains at less than or equal to 64/8 micrograms/ml but failed to suppress the growth of 15 strains producing stably depressed cephalosporinases. Comparisons of piperacillin/tazobactam results determined with 100/10-, 100/20-, and 100/30-micrograms disks established the 100/10-micrograms disk as most usable. Among five different MIC combinations the ratio of eight parts piperacillin to one part tazobactam or fixed concentration tests at greater than or equal to 4 micrograms tazobactam/ml were preferred, each producing very low occurrences (less than or equal to 1.6%) of false-resistance or susceptibility when compared to disk test results. MICs determined by agar and broth microdilution methods were essentially the same. The recommended breakpoints for piperacillin/tazobactam MICs were identical to those now found in the NCCLS susceptibility testing standards with the following exceptions: (1) for tests with H. influenzae and Staphylococcus spp.--susceptible at greater than or equal to 21 mm (MIC less than or equal to 16/2 micrograms/ml) and resistant less than or equal to 20 mm (MIC less or equal to 32/4 micrograms/ml); and (2) all remaining nonspeudomonas isolates would be interpreted by the NCCLS piperacillin enteric bacilli susceptibility criteria. This newer beta-lactamase inhibitor combination appears to be worthy of further in vivo trials guided by these or similar tentative in vitro susceptibility testing parameters. PMID- 2560423 TI - Activity of ticarcillin/clavulanate and piperacillin/tazobactam (YTR 830; CL 298,741) against clinical isolates and against mutants derepressed for class I beta-lactamase. AB - Piperacillin/tazobactam (YTR 830; CL-298,741) was tested against fresh and stock clinical isolates of Gram-negative bacilli, as well as against Gram-negative bacilli that had stably derepressed Class I beta-lactamases or that were hyperproductive of non-Class I beta-lactamases. Of 63 clinical isolates of the Enterobacteriaceae with ticarcillin (plus clavulanate) minimum inhibitory concentrations (MICs) of greater than or equal to 128 micrograms/ml, 16 had piperacillin/tazobactam MICs of less than or equal to 16/2 micrograms/ml. Of 48 clinical isolates of Pseudomonas spp. with ticarcillin (plus clavulanate) MICs of greater than or equal to 128 micrograms/ml, 35 had piperacillin/tazobactam MICs of less than or equal to 64/8 micrograms/ml. Tazobactam generally reduced piperacillin MICs by two- to greater than or equal to eightfold against stably derepressed mutants for Class I beta-lactamases. PMID- 2560425 TI - [Viral etiology of cancer of the uterine cervix. II. Detection of human cytomegalovirus DNA sequence and antigen in cervical lesions]. AB - Human cytomegalovirus (HCMV) DNA sequence and HCMV antigen were detected by dot blot hybridization and PAP staining in biopsy specimens from cervical lesions. The results showed that the positive rate of HCMV DNA sequence was 73% in cervical cancer and 73% in cervical dysplasia which were higher than that in chronic cervicitis (44%) (P less than 0.05). HCMV antigen could be detected in all kinds of cervical lesions. But there was no significant difference in the positive rates. The detection rate of HCMV DNA sequence was not always in accordance with that of HCMV antigen. The results indicate that HCMV may be another high risk factor of cervical cancer. PMID- 2560424 TI - AAQUANT: a computer program for quantitative amino acid analysis of proteins and peptides. AB - Quantitative amino acid analysis is an important tool used in the characterization and structural determination of peptides and proteins. A new computer program, AAQUANT, has been developed specifically to aid researchers in analyzing amino acid composition data. AAQUANT calculates amino acid recoveries, including 95% confidence intervals, following acid hydrolysis of peptides and proteins, and also includes useful routines to locate regions of a specified amino acid composition in known protein sequences, compute amino acid composition reports of known protein sequences, generate proteolytic digestion maps of proteins, and create and edit protein sequence data files. This report describes the AAQUANT routines, and demonstrates the use of the program. PMID- 2560426 TI - [Influence of blood transfusion on postoperative prognosis of stage I (No) lung cancer]. AB - Clinical and basic investigations suggest that perioperative blood transfusions may increase cancer recurrence by nonspecific immunosuppression. Fifty-three patients with Stage I (No) lung cancer treated by surgery in the University of Chicago Medical Center from 1970 to 1986 are reported. There were 18 squamous carcinoma, 29 adenocarcinoma and 6 large cell carcinoma. Of these 53 patients, 30 (57%) received blood transfusion (BT) and 23 (43%) did not (non-BT). A disease free survival rates of BT and non-BT groups were 43.3% (13/30) and 78.3% (18/23), respectively. Mortality rates after recurrence of BT and non-BT groups were 46.7% (14/30) and 8.7% (2/23) (P less than 0.01). Difference in mortality rates of patients who received red blood cells only (30.8%) and multiple blood transfusions (58.8%), was not significant (P greater than 0.05). This study confirms others reports that perioperative blood transfusions may adversely influence the survival of Stage I (No) lung cancer patients. PMID- 2560427 TI - [Synthesis of ATP by the surface of isolated skin of the frog Rana ridibunda]. PMID- 2560428 TI - [Inhibitory effect of caffeine on the potential-dependent calcium current in isolated intestinal smooth muscle cells]. PMID- 2560429 TI - Increased response of cerebellar cGMP to kainate but not NMDA or quisqualate following barbital withdrawal from dependent rats. AB - Female Sprague-Dawley rats were maintained on a diet of powdered food containing barbital for 8 weeks before the drug was abruptly withdrawn. Twenty-four hours later both barbital-dependent and control rats were injected intracerebroventricular (i.c.v.) with saline or one of four doses of kainic acid (KA) or in a separate experiment with saline or one of three doses of N-methyl-D aspartic acid (NMDA) or of quisqualic acid (QA). After 4.5 min, the animals were killed by focused microwave irradiation, and the cerebella were collected. The levels of cyclic guanosine 3',5' monophosphate (cGMP) were markedly elevated in the cerebella of barbital-withdrawn rats when compared to controls. When compared to saline treatment, KA, at all dosages, resulted in a significantly greater elevation of cerebellar cGMP in the barbital-withdrawn rats than was induced by drug withdrawal alone. Only the two higher dosages of KA produced a significant elevation of this parameter in the control rats. Unlike KA, neither QA or NMDA produced any greater elevations of cGMP in barbital withdrawn rats than were induced by drug withdrawal alone. These collective results suggest that there is an increase in the response to KA but not QA or NMDA following the withdrawal of barbital from dependent rats. PMID- 2560430 TI - An in vitro profile of activity for the (+) and (-) enantiomers of spiradoline and PD117302. AB - Kappa- and mu-opioid binding site affinities of the kappa-selective ligand U62066 and its optical isomers, (+)-U63639 and (-)-U63640 were compared with those of the structurally related ligand PD117302 and its respective isomers, (+)-PD123497 and (-)-PD123475. The relative efficacies of each compound were also established using the guinea-pig ileum, rat and rabbit vas deferens smooth muscle bioassays. The specific opioid receptor mediating the agonist behaviour was determined in the guinea-pig ileum bioassay by obtaining pKB values for naloxone and for the kappa-selective antagonist nor-binaltorphimine. Both racemic compounds and the ( )-enantiomers displayed high selectivity for the kappa-receptor with (-)-PD123475 the most selective. The (+)-enantiomer, PD123497, was approximately equipotent at mu-/kappa-sites while (+)-U63639 displayed a 140-fold mu-receptor selectivity. Bioassay studies showed each compound to be interacting at the kappa-receptor, with the exception of (+)-U63639 which displayed a profile consistent with that of a weak mu-receptor agonist. PMID- 2560431 TI - Stereospecific effects of a kappa-opiate antagonist on the actions of morphine in morphine-tolerant rats. AB - The effects of kappa-opiate receptor antagonist, MR 2266 and its dextro isomer, MR 2267 on morphine-induced analgesia and changes in colonic temperature were determined in morphine-naive and morphine-tolerant male Sprague-Dawley rats. Intraperitoneal administration of morphine (8 mg/kg) produced analgesia and hyperthermia in morphine-naive rats. MR 2266 (0.3-3.0 mg/kg) antagonized morphine induced analgesia and hyperthermia in morphine-naive rates but MR 2267 was inactive. Subcutaneous implantation of six morphine pellets during a 7 day period induced tolerance to the analgesic and hyperthermic effects of morphine in the rat. MR 2266 also antagonized morphine analgesia and hyperthermia in morphine tolerant rats, however, MR 2267 had no effect. A high dose (3 mg/kg) of MR 2266 produced an intense hypothermic response in morphine-tolerant rats. Previously we have shown that kappa-opiate receptor agonists antagonize morphine analgesia in morphine-naive rats but potentiate it in morphine-tolerant rats. The results of the present studies indicate that the antagonist of kappa-opiate receptors, on the other hand, antagonize morphine effects in both morphine-naive and morphine tolerant rats in a stereospecific manner. PMID- 2560432 TI - MDL 73005EF: partial agonist at the 5-HT1A receptor negatively linked to adenylate cyclase. AB - MDL 73005EF has been recently described as a potent, highly selective 5-HT1A ligand. Although proposed to act predominantly as an antagonist (M. Hibert, A.K. Mir, G. Maghioros, P. Moser, D.N. Middlemiss, M.D. Trickleband and J.R. Fozard, 1988, The Pharmacological properties of MDL 73005EF: a potent and selective ligant at 5-HT1A receptors, Br. J. Pharmacol. 93, 2P), we have demonstrated that MDL 73005EF also acts as a highly efficacious partial agonist at the 5HT1A receptor, based on its ability to inhibit forskolin-stimulated adenylate cyclase in rat hippocampal membranes. Compared with two structurally related 5-HT1A partial agonists, the rank order of potency of MDL 73005EF in the FSC assay was comparable to affinity calculated by radioligand binding. PMID- 2560433 TI - Characterization of 5-hydroxytryptamine3 receptors in the medial prefrontal cortex: a microiontophoretic study. AB - The microiontophoretic application of the selective 5-hydroxytryptamine3 (5-HT3) agonist 2-methylserotonin suppresses medial prefrontal cortex cell firing. This effect is blocked by the 5-HT3 antagonists BRL 43694 and ICS205930, but not by metergoline or (+/-)-pindolol. Continuous microiontophoretic administration of magnesium chloride or the gamma-aminobutyric acidA antagonist SR 95103 did not alter 2-methylserotonin's suppressant action, suggesting that this effect is direct. Our results suggest that 5-HT3 receptors have a functional role in the medial prefrontal cortex. PMID- 2560434 TI - Enhancement of benzodiazepine receptor binding by L-lysine is chloride-dependent and due to increase in binding affinity. AB - L-Lysine enhanced specific [3H]flunitrazepam binding dose dependently on extensively washed bovine brain membrane in vitro. This enhancement was stimulated by chloride ions dose dependently. Scatchard analysis indicated this enhancement by L-lysine to be due to increase in binding affinity (KD) with no change in receptor density (Bmax). Since enhancement of [3H]flunitrazepam binding by L-lysine was partially inhibited by picrotoxinin, L-lysine may act on a distinct picrotoxinin-sensitive site which was distinct from the gamma aminobutyric acid receptor site. This binding site, however, appears to have some features resembling that of the central nervous system-depressant barbiturates. PMID- 2560435 TI - Non-benzodiazepine, Ag-dependent, brain-specific binding sites for [3H]beta carboline-3-carboxylic acid ethyl ester. AB - Specific, saturable and reversible binding of [3H]beta + CCE ([3H]beta-carboline 3-carboxylic acid ethyl ester) in buffer containing 20 microM AgNO3 and 10 microM diazepam was detected in rat brain membranes. The binding of [3H]beta CCE to non benzodiazepine binding sites is Ag+ (Cu)-dependent, stimulated by NaCl and ascorbic acid and inhibited by dithiothreitol. The concentration of non benzodiazepine [3H]beta CCE binding sites (Bmax) determined in the brain membranes was 1180 +/- 320 pmol/g tissue, and Kd = 77 +/- 19 nM. [3H]beta CCE bound to benzodiazepine receptors in the same membranes with Bmax = 81 +/- 9 pmol/g tissue and Kd = 3.2 +/- 0.4 nM. PMID- 2560437 TI - [Mucoepidermoid cancer of the esophagus with 9 years' follow-up]. PMID- 2560436 TI - [A preclinical evaluation of the antianginal activity of energy metabolism intermediates]. AB - In acute experiments on dogs and cats with occlusion of the coronary arteries the antianginal activities of malate (100 mg/kg) and NAD (0.2 mg/kg) were studied. The drugs were found to increase the collateral blood flow and the contractile activity of the myocardium against the background of a moderate rise of the heart oxygen consumption and to normalize the systemic hemodynamics parameters. PMID- 2560438 TI - [Treatment of minor hepatic tumors with ultrasonically guided percutaneous injection of absolute alcohol. Results in 8 patients]. AB - The effectiveness of ultrasonically guided percutaneous ethanol injection therapy was investigated in 8 patients who had hepatocellular carcinoma (two cases), hepatic metastases from colorectal carcinoma (five cases) and hepatic metastasis from breast carcinoma (one case). Tumor diameter was less than 5 cm in all cases. Fifty-nine ethanol injection were performed for 11 lesions. Histopathologic examination, performed in all cases, showed that the tumor was completely necrotic in 4 cases, partially necrotic in 3 cases and unchanged in 4 cases. Serum alphafetoprotein and carcinoembryogenic antigen decreased in 5 out of 8 cases. Seven patients were still alive at the end of the study with a mean follow up period of 10.7 months. We conclude that ethanol injection may be a valuable treatment for small hepatic tumors. PMID- 2560439 TI - [Recurrence of the initial disease after liver transplantation]. PMID- 2560440 TI - [Characteristics of structures of Drosophila polytene chromosomes formed by transposable DNA fragments]. AB - An electron microscopic analysis of regions of Drosophila melanogaster polytene chromosomes into which DNA fragments of different genetic composition were inserted by the P element-mediated transformation was performed. In 4 of 5 regions studied with integrated DNA sequences of the hsp28-ry, hsp70-Adh, ry hsp70-beta-gal genes new bands appeared. Apparently their generation is mainly caused by integration of the DNA fragments in interbands. Absence of a new band in transformed region in one of the stocks can be explained by fusion of the insertion with a band existed in the initial untransformed stock. Among the transformants studied, the minimum length of DNA fragment revealed as a new band is about 5 kb. DNA packing ratio of such the bands varies from 30 to 50. The activation of the inserted genes by heat shock allows to trace peculiarities of the new bands puffing. The puff sizes correlate with the length of the activated genes. If the DNA of the fragment consists of the sequence of one gene, its activation will lead to decondensation of the whole band. In the case when DNA fragment consists of 2 genes and the promoter of activated gene is situated inside the sequence, the band is splitted after gene activation at the beginning and then separated portion of the band is decondensed and puffed. The data obtained evidence that a band of polytene chromosome is not a unit of decondensation. DNA packing ratio in puffs is equal to 1.5-3.5. PMID- 2560441 TI - Atypical presumed CMV retinitis. AB - Cytomegalovirus (CMV) retinitis is a common and one of the most serious ophthalmic manifestations of the acquired immunodeficiency syndrome (AIDS). Early recognition is essential in the proper management of this necrotizing and progressive infection. We discuss an unusual presentation of CMV retinitis which appeared initially only in the foveal region. PMID- 2560442 TI - [Use of azopyram for determining the peroxidase level in monitoring the quality of thermal treatment of meat products]. PMID- 2560443 TI - Constellations of genetic abnormalities predict clinical outcome in childhood malignancies. PMID- 2560444 TI - Cytostatic influence of thioproline on peripheral lymphocytes of healthy persons and non-Hodgkin's lymphoma patients. PMID- 2560445 TI - Signal transduction mechanisms in human natural killer cells mediating antitumor immunity. PMID- 2560446 TI - Modification of HLA expression as a possible factor in the pathogenesis of Burkitt's lymphoma. AB - Our data show that: Most BL cells express significantly less HLA class I and II than the corresponding LCLs. Lymphoblastoid cell lines have a much higher HLA class I and II density than normal peripheral blood B cells. There are qualitative differences in class I and II expression in most BL/LCL pairs. These qualitative differences are not caused by EBV infection. They might be due to a selective downregulation of HLA specifities by c-myc activation. We conclude that these modifications of HLA expression may well play a role in the reduced specific immune recognition of Burkitt's lymphoma cells. PMID- 2560447 TI - Isolation and characterisation of a myeloid leukaemia inducing strain of feline leukaemia virus. PMID- 2560448 TI - HIV-I replication requires an intact integrase reading frame. PMID- 2560449 TI - Even transcriptionally competent proviruses are silent in bovine leukemia virus induced tumor cells. PMID- 2560450 TI - [Conservative treatment of early tubal pregnancy]. AB - Due to improved diagnosis most tubal pregnancies are detected today before tubal rupture. Therefore various conservative treatment modalities have been described lately. In the present study 73 cases of unruptured tubal pregnancies are described that where treated with local and systemic PG application. With an initial beta-hCG level of below 2,500 mIU/ml serum, 45 of 53 cases could be treated successfully without classical operation. With an initial beta-hCG above 2,500 mIU/ml serum the results were less satisfactory. Hysterosalpingography controls showed that almost all pertinent tubes are patent after such treatment. PMID- 2560451 TI - [The significance of human papillomavirus infection in cervix cancer and its preliminary stages (including other HPV-induced genital lesions)]. AB - Southern blot analysis, DNA dot blot technique and in situ hybridization were used to examine cells and tissues from lesions of the female genital tracts for infections of human papillomaviruses (HPV) of the types 6/11, 16/18 as well as 31/33 and 35. As the degree of dysplasia increased the proportion of HPV-positive results increased significantly, especially for the HPV types 16/18; there was a decrease in the HPV DNA 6/11-positive cases. In the first follow-up examinations of cervical smears of the Pap. IIID type after 6 months there was a persistence of 84.2%. Of these, 57.8% were HPV 16/18-positive, and 18.2% could be classified as belonging to Pap. group IV. The importance of HPV typing lies in the definitions of the pathogens, risk groups, therapeutic measures and their controls. PMID- 2560452 TI - Molecular basis of red cell enzymopathies associated with hereditary nonspherocytic hemolytic anemia. AB - In the past few years, very rapid advances have been made in the field of red cell enzymopathies associated with hereditary nonspherocytic hemolytic anemia, particularly in molecular basis. Nucleotide sequence and amino acid sequence of normal human red cell enzymes have been clarified in phosphofructokinase, aldolase, triosephosphate isomerase, phosphoglycerate kinase, pyruvate kinase, diphosphoglycerate mutase, glucose 6-phosphate dehydrogenase, adenylate kinase and adenosine deaminase. Furthermore, in aldolase-, triosephosphate isomerase-, diphosphoglycerate mutase-, glucose 6-phosphate dehydrogenase-, and adenylate kinase deficiency, single nucleotide changes which cause single amino acid substitutions and finally hemolysis, have been found. PMID- 2560453 TI - [Ganciclovir (DHPG)--treatment of cytomegalovirus retinitis in AIDS]. AB - The clinical course of 3 patients treated with the virostatic drug dihydroxypropoxymethylguanine (DHPG) is reported. With maintenance therapy the progression of retinitis was slow, but was not stopped. More rapid progression of retinitis could be observed after cessation of therapy because of DHPG-induced leukopenia. Therefore, consequent maintenance therapy should be the therapeutic aim. Under maintenance therapy, useful visual acuity could be maintained until death. PMID- 2560454 TI - Different effects of aging on the opioid mechanisms controlling gonadotropin and cortisol secretion in man. AB - The present study was undertaken in order to assess the influence of aging on the endogenous opioid control of gonadotropin and adrenocorticotropin/cortisol secretion in man. For this purpose, the capability of the opioid antagonist naloxone to increase circulating levels of luteinizing hormone (LH), follicle stimulating hormone (FSH) and cortisol was tested in male subjects of different ages. Thirty normal men were randomly chosen and divided into 3 groups by age: group I = 22-40 years (n = 10); group II = 41-59 years (n = 10); group III = 62 80 years (n = 10). Since the men of group III showed higher basal serum gonadotropin concentrations than the subjects of group I and group II, we selected from a large population a fourth group of elderly men with normal basal LH and FSH levels: group IV = 61-82 years (n = 7). All subjects were tested for 120 min during the intravenous administration of naloxone (4 mg given in an intravenous bolus at time 0, plus 10 mg infused for 2 h). Control tests with normal saline instead of naloxone were performed in all groups. All subjects had similar blood testosterone and cortisol levels, whereas LH and FSH concentrations were significantly higher in group III than in groups I, II and IV. Naloxone increased plasma cortisol concentrations by 50% in all groups. The cortisol secretory response followed a similar pattern regardless of age.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2560455 TI - Developmental pattern of 17 beta-hydroxysteroid dehydrogenase and 5 alpha reductase activities in the foreskin of boys from birth to eight years of age. AB - The normal developmental pattern of 17 beta-hydroxysteroid dehydrogenase (17HSD) activity in genital skin was examined using radiolabeled androstenedione as a substrate in a microassay based on high-pressure liquid chromatography separation of the metabolites. This assay allowed the simultaneous determination of 17HSD and 5 alpha-reductase (5R) activities in both individual foreskin samples and pools of tissue obtained at circumcision from birth to 8 years of age. The results show that 17HSD activity is very low at birth and increases steadily during the so-called quiescent period. Reciprocal changes were observed for 5R. The increase in 17HSD activity appears to be independent of gonadal stimulation, but the mechanisms involved remain to be elucidated. From a clinical standpoint, our results provide an alternative explanation for the relative lack of virilization observed in newborns with testicular 17HSD deficiency. PMID- 2560456 TI - Relationship of parathyroid adenoma volume and biochemical function. AB - We retrospectively studied data of 24 patients with surgically proven hyperparathyroidism to determine whether biochemical function correlated with the calculated volume of parathyroid adenoma. Carboxyl-terminal parathyroid hormone (C-PTH), amino-terminal parathyroid hormone (N-PTH), nephrogenous cyclic AMP (N CAMP), and other markers of calcium metabolism were measured with standard techniques. Tumors were divided into small (less than 1.0 cm3) or large (greater than or equal to 1.0 cm3) sizes. N-PTH and C-PTH measurements were increased in 10% and 30% of patients with small tumors and in 71 and 78% of patients with large tumors. Serum calcium, N-CAMP, C-PTH and N-PTH tended to be greater with large tumors, but only C-PTH was significant. We concluded that the size of parathyroid adenoma influenced biochemical measurements of its activity and that the measurements of PTH in patients with small tumors are not as sensitive as those in patients with large tumors. PMID- 2560457 TI - [Amplification, rearrangement and over-expression of c-fos gene in human primary brain tumors]. AB - The c-fos gene was used as a probe to detect the Bam HI-digested brain DNA and total RNA isolated from 2 cases of normal human brain and 11 cases of human brain tumor by Southern blot analysis and RNA dot hybridization technique. The result showed an amplification and over-expression of c-fos gene in one case of glioblastoma multiforme. These data suggest that the c-fos gene may take an important role in the carcinogenesis of human primary brain tumor, and the level of c-fos expression may be correlated with the degree of differentiation of brain tumor cells. We also found that there was a rearrangement of c-fos gene in one case of ependymoma. This suggests that, in ependymoma, the c-fos gene may be activated in a way different from that in the brain tumors of astroglia origin. PMID- 2560458 TI - [Study on molecular hybridization with biotin-labelled HPV 16 DNA probe in human cervical carcinoma]. AB - Biotin-labelled human papillomavirus (HPV) 16 type DNA probe was prepared by the techniques of molecular biology. And dot hybridization technique was used to detect the HPV 16 homologous sequences in the tissues DNA of human cervical carcinoma. The results indicated that 16 cases out of 28 of the human cervical carcinoma tissues were positive. The positive rate was 57%. The other 4 cases of normal uterine cervix tissues were negative. Only 1 in 4 chronic cervicitis tissues showed positive. The HPV 16 plasmid DNA, as the positive control group, showed strong positive, while lambda-phage DNA was negative. The results have shown that the genome of the HPV actually exists in the tissue of the cervical carcinoma and that there is a close relationship between the cervical carcinoma and HPV infection. This experiment adopted the Biotin-labelled HPV 16 DNA probe. And it may provide us with a quick and sensitive method for investigation of the infection of HPV and its role in the carcinogenesis of cervical carcinoma. PMID- 2560459 TI - [Study on identification of liver isoferritin by isoelectrofocusing in thin-layer polyacrylamide gels]. AB - Thin-layer IEF, an analytical technique of high resolution for protein, was used to identify the liver isoferritin in Wistar rat, normal human and patients with hepatocellular carcinoma (HCC) respectively. The focusing conditions were 2000 V, 25W, 10 degrees C, 2h. The results showed that the IEF bands of the liver isoferritin in Wistar rat, normal human and patients with HCC were 7, 6, 3 and the pI of the liver isoferritin were 5.76-4.75, 5.80-4.85, 4.50-4.43 respectively. The method and data in this paper might be good reference for in depth study of the structure and function of ferritin and its applications in clinical medicine. PMID- 2560460 TI - [Primary study of histogenesis and diagnosis of synoviosarcoma]. AB - Fourteen cases of synoviosarcoma were investigated by electron microscopy and immunohistochemical technique (PAP) using keratin and EMA as epithelial markers, vimentin as a mesenchymal marker. Of the 14 cases 9 were biphasic (BS) and 5 were monophasic synoviosarcoma (MS). In BS, epithelial-like cells, glandular, tubular of slit-like structures, junctional complex, microvilli at the luminar surface and basal lamellae at bottom could be easily seen. Spindle cells both in BS and MS were similar. These cells had somewhat similarity with epithelia-like cells, such as slender cell processes or microvilli, the external lamellae and cell junctions. The spindle cells also formed some slit-like structures. Collagenization was mild in matrix. Whether in BS or MS, epithelial or spindle cells, there were positive reactions for keratin, EMA and vimentin. The present study suggests that synoviosarcoma is neither a tumor of synovium origin nor a sarcoma of synovial differentiation, but a carcinosarcoma or adenosarcoma of soft tissues; the monophasic type is not a variant of fibrosarcoma either. It really is a distinct variant of synoviosarcoma. The expressions of both epithelial and mesenchymal features are useful criteria for diagnosis of synoviosarcoma. PMID- 2560461 TI - A major role of macrophage activation by interferon-gamma during mouse hepatitis virus type 3 infection. I. Genetically dependent resistance. AB - Resistance of mice to mouse hepatitis virus type 3 (MHV3) infection is genetically determined. Normal adult A/J mice are resistant, and BALB/c mice are susceptible. Higher titers of virus and interferon (IFN) in vivo were found in MHV3-infected BALB/c mice compared with A/J mice. In vitro activation of macrophages (M phi) by lipopolysaccharide (LPS) delayed MHV3 replication only in cells that originated from A/J mice, although cell populations from both A/J and BALB/c mice were able to synthesize comparable amounts of IFN-alpha/beta. Using specific antibodies, we have shown that the delayed MHV3 replication in LPS activated A/J M phi was due, in part, to IFN-alpha/beta. A/J M phi were found to be more sensitive to IFN-gamma than to IFN-alpha/beta, and BALB/c M phi did not develop an antiviral state to either IFN. Cultured spleen cells from A/J mice synthesized more IFN-gamma than BALB/c spleen cells after specific or non specific stimulation. The results indicate that IFN-activated M phi may play a crucial role in the resistance to MHV3 infection. Since IFN-gamma is produced in large amounts by A/J spleen cells after specific stimulation with MHV3 and is efficient in activating the A/J M phi, a T cell-dependent mechanism is likely to be involved. PMID- 2560463 TI - Triggering of neutrophil cytotoxicity against an antibody-coated tumour target by TPA. AB - The human erythroid myeloid leukaemia cell line K562 was used as target for human neutrophil cytotoxicity. Neutrophils demonstrated cytotoxicity against K562 only in the presence of a second stimulus, tetradecanoyl phorbol acetate (TPA), a result consistent with previous observations. We now demonstrate that antibody coated K562 (using OKT9 and 345 monoclonal antibodies) are similarly only sensitive to neutrophils when TPA is added. The presence of both antibody and TPA in the cytotoxic assay resulted in significantly higher levels of cytotoxicity than in the absence of antibody; the result being consistent with a synergistic action between protein kinase C activation and Fc receptor perturbation in the neutrophil. The cytotoxicity against non-coated and antibody-coated targets was markedly inhibited, particularly against the former, by the protein kinase C inhibitor, 1-(5-isoquinoline-sulfonyl)-2-methyl piperazine (H-7). There were marked differences in the extracellular calcium dependency of the two types of cytotoxicity reactions. TPA-activated respiratory burst was unaffected by the presence of non-coated and OKT9-coated targets, whereas TPA-induced lysosomal enzyme release was significantly increased by non-coated targets and a further increase occurred in the presence of OKT9-coated K562. PMID- 2560464 TI - The generation and activation of memory class I MHC restricted cytotoxic T cell responses to influenza A virus in vivo do not require CD4+ T cells. AB - The cellular requirements for the generation and activation of anti-influenza memory class I MHC restricted Tc cell responses were studied by selectively reconstituting lethally irradiated mice. The generation of memory Tc cells was investigated by using unprimed splenocytes to reconstitute infected, lethally irradiated mice; the activation of memory Tc cells was tested by using primed splenocytes to reconstitute uninfected, lethally irradiated mice. It is shown here that depletion of CD4+ T cells from donor cells did not reduce Tc cell responses in recipient mice. Depletion of CD8+ T cells from donor splenocytes prevented the memory Tc cell responses. Thus depletion of CD4+ T cells had no effect on the generation and activation of the memory Tc cells in vivo. PMID- 2560462 TI - Involvement of reactive oxygen metabolites in the candidacidal activity of human neutrophils stimulated by muramyl dipeptide or tumor necrosis factor. AB - In the presence of the adjuvant glycopeptide muramyl dipeptide (MDP), purified human PMN exhibited an enhanced capacity to kill Candida albicans cells at various cell ratios. A significant effect was obtained at 100 ng/ml MDP, and the maximum was reached at 1 micrograms/ml MDP. Recombinant human tumor necrosis factor (rHuTNF), a monokine that enhances host resistance to bacterial and fungal infections, also stimulated the candidacidal potency of PMN with a maximal effect at 10(-2) ng/ml rHuTNF. When MDP- or rHuTNF-stimulated PMN were cultured with yeast cells, the intracellular production of oxygen metabolites was enhanced. Pretreatment with inhibitors of oxidative burst demonstrated that the yeast cell killing by MDP-stimulated PMN was not affected by SOD but was inhibited by sodium azide, indicating the involvement of myeloperoxidase (MPO)-halide system in fungicidal mechanisms induced by MDP. When PMN were stimulated with rHuTNF, the killing of yeast cells was neutralized by iodoacetamide, showing that the candidacidal potency of stimulated-PMN was due to oxygen derivatives. Inhibition by sodium azide and sodium benzoate indicated that these oxygen metabolites could be derived from the MPO-halide system but also from hydroxyl radical production. Moreover, SOD partially inhibited the fungicidal potency of rHuTNF-stimulated PMN, thus indicating a possible reutilization of the released O2- anion for intracellular killing. Cytochalasin B abrogated the PMN fungicidal potency in all cases. PMID- 2560466 TI - [Recent studies of the incidence of IgG allo-antibodies in blood transfusion recipients]. PMID- 2560465 TI - Enhancement of u.v.-induced skin carcinogenesis in the hairless mouse by inoculation with cell-free extracts of skin tumours. AB - The presence of papillomaviral-like DNA has been described in ultraviolet light induced tumours in the skin of the hairless mouse (14). Here we describe the effects of the inoculation of cell-free extracts of ultraviolet light-induced tumours into the scarified skin of normal hairless mice, prior to exposure of the mice to a cumulative carcinogenic dose of ultraviolet light. Extracts from papillomas or squamous cell carcinomas enhanced the susceptibility of the inoculated mice to ultraviolet light-induced tumorigenesis, if the extracts contained papillomaviral DNA sequences detected by cross-hybridization with Mastomys natalensis papillomaviral DNA. The recipient mice developed a greater tumour incidence, tumour yield, tumour diameter and degree of malignancy. PMID- 2560467 TI - Thermogenesis and brown fat: relevance to human obesity. AB - Brown adipose tissue (BAT) is a specialized thermogenic tissue, which is highly vascularized and richly innervated with sympathetic nerves. Due to the high thermogenic capacity (500 W/kg) even very small quantities such as those found in adult man can significantly influence het production. As little as 50 g BAT could make a contribution of 10-15% to energy turnover in man. This would be more than sufficient to cause large differences in fat deposition between individuals with active or inactive dietary induced thermogenesis. Recent research has revealed the presence of an atypical beta-adrenoreceptor on BAT, tentatively designated as beta 3-adrenoreceptor. The development of beta 3-agonists offers an opportunity to treat obesity without the cardiovascular and other undesirable side-effects of conventional adrenergic agonists. PMID- 2560468 TI - Hormonal regulation of a rat seminiferous tubule factor which inhibits LH action on interstitial cells. AB - The cytosol from rat testes or seminiferous tubules contains a factor that markedly reduces the responsiveness of interstitial cells to stimulation by LH. It was noted previously that the inhibitor cannot be found until 35 days of age, suggesting that gonadotrophic stimulation of the testes is of importance for its formation. In the present studies, treatment of intact 20-day-old rats with FSH or with a combination of FSH and LH caused a premature appearance of the inhibitory activity. LH alone had a weak effect. However, hypophysectomy at 20 or 35 days of age did not influence the inhibitor content of the testes. Moreover, when the Leydig cells of adult rats were destroyed selectively by treatment with ethylene dimethane sulphonate, inhibitor levels were unchanged. It is suggested that induction of the Leydig cell inhibitor is under the control of FSH. However, once induced, its regulation seems to be independent of the pituitary gland. In separate experiments, ligation of the efferent ducts of the testes in adult animals did not cause any accumulation of inhibitory activity in the ligated testes, nor could the inhibitor be traced in the caput epididymis. Thus, it does not seem to be secreted into the epididymis, but rather may act as a paracrine factor in the testis. PMID- 2560469 TI - Fibrous histiocytomas of the soft palate. AB - Fibrous histiocytomas (FHs) of the oral cavity are exceedingly rare. We report a case of fibrous histiocytoma located in the soft palate of an 11-year-old male. A review of the English literature shows only 4 previously reported cases in this location. This report includes an overview of the clinical and histopathological features of this fibrohistiocytic disorder. A review of the current classification, as well as the diagnosis and management of these neoplasms as they pertain to the head and neck region are discussed. PMID- 2560470 TI - Prophylaxis of fatal pulmonary embolism in general surgery using low-molecular weight heparin Cy 216: a multicentre, double-blind, randomized, controlled, clinical trial versus placebo (STEP). STEP-Study Group. AB - The effectiveness of low-molecular weight heparin CY 216 in the prophylaxis of fatal pulmonary embolism in patients undergoing general surgery was assessed in a multicentre, double-blind, randomized, clinical trial against placebo. A total of 4,498 patients aged over 40 undergoing general surgery were enrolled in the 18 centres which took part in the trial. Patients received a single daily subcutaneous injection of 7,500 anti-Xa units I.C. of CY 216 or placebo two hours before surgery, 12 hours after the initial injection and then daily for at least seven days. A post-mortem examination had to be carried out in every patient who died. The two groups of patients were well-matched for age, sex, type of disease, site and duration of operation as well as for incidence of risk factors which could predispose to the development of thromboembolism. Twenty-six deaths were recorded and validated: eight (0.36%) in the CY 216 group and 18 (0.80%) in the placebo group (p less than 0.05). At the post-mortem examination, carried out in 23 patients (88.5%), two deaths were found to be directly due to pulmonary embolism (0.09%) in the CY 216 group and four (0.18%) in the placebo group. Pulmonary embolism contributed to death in four other placebo-treated patients. Pulmonary or extrapulmonary thromboembolism was a significantly less frequent direct cause of death (p less than 0.05) in the CY 216 group (two pulmonary embolisms) than in the placebo group (four pulmonary embolisms, one acute myocardial infarction, one disseminated intravascular coagulation, two ischemic cerebral strokes).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2560471 TI - A biometrical study on esophageal invasion of carcinoma of the upper stomach. Part I: Determination of occurring point of the cancer and analysis of its horizontal extension. AB - In order to detect the occurring point of gastric cancer, and to determine its horizontal extension from this point, we performed a histological examination of 30 resected specimens of upper stomach cancer and 31 of middle and lower stomach cancers. The center of the horizontal extension (Oh) and that of the vertical extension (Ov) were calculated using a paper model. The mean distance between Ov and Oh (Ov-Oh D) was as little as 5.7 +/- 5.0 mm, and the ratio of Ov-Oh D to the size of the lesion was as low as 8.3 +/- 6.7%. There was a linear correlation between the oral and anal radii of the cancer. There was also a linear correlation between anterior and posterior radii of the cancer. In conclusion, either the Oh or the Ov can be considered as the occurring point, and the cancer grows concentrically around this point. PMID- 2560473 TI - Characterization and zeste reaction of spontaneously reduced transposable elements in Drosophila melanogaster. AB - Seven transposable elements (TE), selected from parental stock w 239 Cy, have been characterized. They all carry w+ but have lost both rst+ and the two polytene chromosome bands of a complete TE and are thus cytologically invisible. In situ hybridization with probes from the white-roughest interval shows that the reduced TE's are truncated in the same region, 40-90 kb upstream of the white locus, well proximal to the zeste interaction site. They express the white gene in wild type fashion in all tissues studied (eyes, testis sheaths, and Malpighian tubules) but give zeste reactions varying from yellow to bright red. Thus, by the criterion of the zeste reaction, 40 kb of TE sequences 5' to the transcription start site cannot insulate the white gene from position effects. One single spontaneously reduced TE (TE203) originated from parental stock w TE100 ct. This TE is truncated closer to the white locus (+ 18- +26 kb), but still well separated from the zeste interaction site. In a z1w11E4 background, homozygous TE203 flies have an uneven eye pigmentation, while heterozygotes have eyes that are yellowish red. The Malpighian tubules and testes show wild type pigmentation in both cases, like normal TE's. PMID- 2560472 TI - Colorectal cancer in young patients. AB - Colorectal cancer is rare in patients under 40 years of age. We reviewed all colorectal cancer patients treated at the First Department of Surgery of the University of Rome, to determine the true incidence of large bowel cancer in young patients. During the period 1978-1984 a total of 571 patients were indexed. Out of these 44 were 39 years of age or less. A group of 527 patients 40 years of age or over served as a control. Neoplasms were localized in the right colon in 19 cases and in the left colon in 25 patients, in the young group. In the control group 178 patients presented neoplasms localized in the right colon, while 349 presented tumors localized in the left colon. According to tumor advancement in the young group tumor classification from A stage tumor through D stage was: 4, 6, 14, 5, 9, 6. In the control group 22 patients presented A stage tumors, 106 B1, 139 B2, 50 C1, 118 C2, 92 D tumors. The operability rate was 93.2% in the young group, and 92% in the old one. The operative mortality rate was 0% in the young group, and 2.5% in the old one. Four complications occurred in the young patients, and 57 in the old patients. The five years survival rate was 56.7% and 52.1% in young and old patients respectively. The results of our study indicate that there are no significant differences in prognosis and five years survival in young patients with colorectal cancer. PMID- 2560474 TI - Preparative purification of functional bacteriorhodopsin by high-performance size exclusion chromatography. AB - High-performance size-exclusion chromatography (HPSEC) was used to produce stable bacteriorhodopsin essentially free of native lipids. The purified bacteriorhodopsin was shown to be highly functional when reconstituted into phospholipid vesicles. Purple membrane was washed in the detergent 3-[( 3 cholamidopropyl)dimethylammonio]-2,2-hydroxy-1-propanesulfonate (CHAPSO) to remove a large fraction (65%) of the membrane lipids, solubilized in Triton X-100 and purified on a Bio-Sil TSK G3000SW column using a CHAPSO mobile phase. Pooled column fractions of bacteriorhodopsin from 25-mg sample loads show a 280/548 nm absorbance ratio of 1.5-1.6 and contain less than 4% endogenous lipids. This HPSEC method requires much less expensive synthetic detergent and is much faster than open column methods [cf. L.J.W. Miercke, P.E. Ross, R.M. Stroud and E.A. Dratz, J. Biol. Chem., 264 (1989) 7531-7535]. PMID- 2560475 TI - Determination of cyclic nucleotide phosphodiesterase activity in cellular systems by ion-pair reversed-phase liquid chromatography. PMID- 2560476 TI - Effects of continuous administration of 1,25-dihydroxyvitamin D3 on plasma minerals and unoccupied colon mucosal 1,25-dihydroxyvitamin D3 receptor concentrations. AB - Six mature nonlactating, nonpregnant Jersey cows were implanted with Alzet mini osmotic pumps, which delivered 50 micrograms of 1,25-dihydroxyvitamin D3 [1,25 (OH)2D3] each day for 7 d in an effort to mimic plasma concentrations of 1,25 dihydroxyvitamin D [1,25-(OH)2D] observed in cows at parturition. Plasma samples were obtained daily beginning 6 d prior to implantation and ending 8 d after removal of the implants. Six biopsies of the descending colon mucosa were obtained per rectum before and after implantation and assayed for unoccupied 1,25 (OH)2D3 receptor concentration. Plasma concentration of 1,25-(OH)2D increased from 37 pg/ml pretreatment to 294 pg/ml with the pumps implanted. Plasma Ca concentration increased within 2 d after implantation and remained elevated for 7 d after the pumps were removed. Unoccupied colon mucosa 1,25-(OH)2D3 receptor mean concentration prior to treatment was 14.6 fmol/mg protein and increased within 2 d following implantation to 30.5 fmol/mg protein. These data suggest that 1,25-(OH)2D3 upregulates its own receptor in the intestine of the cow. PMID- 2560477 TI - Improved energy prediction equations for dairy cattle rations. AB - The objective was to develop equations that would accurately predict energy contents of rations commonly fed to milking cows and that were simple to apply. Six published equations that predicted TDN from ration content of crude fiber or ADF or predicted digestible energy, metabolize energy, or NE(1) from ration ADF content were modified to remove bias caused by dietary fiber contents. The modified equation that predicted metabolizable energy from ADF was tested for accuracy using data from several feeding trials in which DM intakes and dietary ADF contents were known. Dietary metabolizable energy content calculated by the equation was multiplied by DM consumed to obtain metabolizable energy intake. Requirement of dietary metabolizable energy was calculated using factors for maintenance, milk produced, and BW changes from 1988 NRC feeding standard. Concurrence values (energy intake/energy required) from feeding trials were calculated and served as the criterion of equation accuracy. Several trials resulted in concurrence values approximating 1.0, indicating that the equation to predict dietary metabolizable content was accurate. However, values for individual trials ranged from .84 to 1.21. Concurrence values that deviated considerably from 1.0 occurred most frequently in trials in which animals experienced negative or large changes in BW. It appeared that errors may have occurred in calculating energy requirements when BW changes were negative or large. It was concluded that the modification improved the original equation and it probably was accurate. The other modified equations would be expected to be equally accurate. PMID- 2560478 TI - Effect of dietary polyamines and alpha-difluoromethylornithine on regulation of intestinal Na+/H+ exchanger. AB - Polyamines are compounds required for initiation of rapid cellular growth and differentiation in many cell types. Ornithine decarboxylase is the rate limiting enzyme in polyamine synthesis. Fasting and refeeding regulates the activity of ornithine decarboxylase and polyamine content in the intestinal tract. We tested the hypothesis that polyamines regulate cell growth via the Na+/H+ exchanger which is believed to be intimately involved in cell growth. Ileal Na+/H+ activity was therefore examined in control, fasted, refed fasted, and in rats given the specific inhibitor of ornithine decarboxylase alpha-difluoromethylornithine. A well-validated ileal brush border membrane vesicles for the study of Na+/H+ exchange activity was utilized. Fasting markedly decreased while refeeding stimulated Na+/H+ exchange activity at all times studied (P less than 0.05 0.001). Maximal uptake of Na+ at 5 min was 3.12 +/- 0.05, 2.5 +/- 0.05 and 2.22 +/- 0.05 nmol/mg protein in refed, control and fasted rats respectively. Kinetics of amiloride sensitive Na+/H+ exchanger showed a Vmax of 17.1 +/- 3.5, 8.0 +/- 0.64 and 4.7 +/- 1.1 nmol/mg protein per 5 s in refed fasted, control and fasted rats respectively Km values were not significantly different between the groups studied. 2% alpha-difluoromethylornithine given in the drinking water abolished the stimulation in Na+/H+ exchange activity in refed fasted rats. These results suggest a close relationship between polyamines and Na+/H+ activity in the intestinal mucosa of rats. PMID- 2560479 TI - Viral contamination of some drinking waters in Romania: a twenty years survey. AB - A synthesis is made on the 20 years virological survey of the drinking water from some towns of Romania. The sampling of the water was made by the gauze-pad method. The virus concentration method by adsorption-elution with the yeast cells was applied concomitantly, at first with the PE60 method, then, during the last years, either with aluminum hydroxide or with the polymer PV methods, and the concentrates were inoculated both into suckling mice and into cell cultures. Various types of coxsackievirus A and B, poliovirus and adenovirus were detected. The proportions of positive samples varied between 0 and 25% annually, in diverse towns, the mean proportion being 2.1%. These proportions are relative low, although two concentrating and two detecting methods were applied concomitantly. PMID- 2560480 TI - [Morphological approach to the neurotransmission system]. PMID- 2560481 TI - [Basic and clinical aspects of ACTH]. PMID- 2560482 TI - Increased N-myc mRNA expression associated with dibutyryl cyclic AMP induced neuroblastoma differentiation. AB - The N-myc cellular oncogene is frequently amplified and expressed at a high level in neuroectodermal tumor cells such as neuroblastoma and retinoblastoma. We examined N-myc expression in NCB-20 hybrid (N18TG2 neuroblastoma x embryonic Chinese Hamster brain) cells. After five days of culture, cells treated with 1 mM db cAMP show extensive neurite outgrowth and secrete acetylcholinesterase into the media at a level three times higher than untreated control. In situ hybridizations, dot blots, and Northern analyses reveal four- to eight-fold higher levels of N-myc mRNA in the treated, differentiated cells than in the untreated, undifferentiated controls. Our results show that the highly differentiated state is not incompatible with a high level of N-myc mRNA. PMID- 2560483 TI - Ca2(+)-dependent proteolytic modification of the cAMP-dependent protein kinase in Drosophila wild-type and dunce memory mutants. AB - Two cAMP-dependent protein kinases with different activation constants were separated from Drosophila melanogaster head extracts. Both are only found in nervous tissue. The first cAMP-dependent kinase, with Mr = 190,000, has been already characterized as tetrameric Drosophila type II cAMP-dependent protein kinase R2C2. The second purified cAMP-dependent protein kinase, with Mr = 80,000, is dimeric in structure RPC, and the cAMP-concentration required for half maximal activation is 4 fold lower than for the type II kinase. The generation of RP can be stimulated in vitro by addition of exogenous calcium and is due to an endogenous Ca2(+)-dependent protease that selectively degrades the regulatory subunit. Extraction in the presence of various protease inhibitors does not affect the amounts of RP, suggesting that the observed quantitative change in RP occurs in vivo. The amounts of RP in the nervous tissue of the memory mutants dunce1 and dunce2, which have increased cAMP levels, are different from the amount of RP in wild-type flies. Also treatments of wild-type flies with drugs affecting cAMP-metabolism and acetylcholine levels led to amounts of RP different from untreated flies. PMID- 2560484 TI - Microbial synthesis of metabolites with antihypertensive activity: aspects of fermentation derived inhibitors of angiotensin-converting enzyme (ACE). AB - In this review the microbial angiotensin-converting enzyme inhibitors are described. Especially from the microbiological point of view the characteristics of these metabolites are given, e.g. occurrence, fermentation physiology and specificity. Besides these data, the structure, assays and some isolation problems are summarised. Apart from ACE inhibition the different biological activities of these secondary metabolites are discussed. PMID- 2560485 TI - Pseudothrombophlebitis in neuropathic arthropathy. AB - The pseudothrombophlebitis syndrome is a complication of large, ruptured, or dissecting popliteal cysts. We report a patient with neuropathic arthropathy of the knee and the pseudothrombophlebitis syndrome secondary to a dissecting popliteal cyst. This represents another form of arthritis associated with this syndrome. PMID- 2560486 TI - [Endoscopic study on the effect of dietary fiber against 1,2-dimethylhydrazine induced colonic carcinogenesis in rats]. AB - The effect of dietary fiber on colonic carcinogenesis was studied endoscopically in rats administered 1,2-dimethylhydrazine. Colonic tumors appeared about six weeks later in rats fed 15% cellulose diet or 40% wheat bran diet than in those fed basal diet. Furthermore, the incidence of colonic tumor was significantly lower from 23th week to 26th week in the cellulose diet group (p less than 0.01) and at 26th week in the wheat bran diet group (p less than 0.05) than in the basal diet group. However, the incidence of colonic tumors and the mean number of tumors per rat at sacrifice (30th week) were not significantly different between these two high-fiber diet groups and the basal diet group. These two diets significantly increased feces 3 to 4 times in weight and 5 times in volume over the base diet. However, 15% pectin diet could neither inhibit colonic carcinogenesis nor increase the fecal weight. These results suggested that cellulose and some dietary fibers of wheat bran were effective to retard colonic carcinogenesis, and that the increased fecal weight and volume by fibers may be involved in the inhibitory effect. PMID- 2560487 TI - [Circulating mature NK (Leu11+) cells positive for IL2 receptor and HLA-DR in patients with various liver diseases and asymptomatic HBsAg carriers]. AB - IL2R+ Leu11+ cells (A) and HLADR+ Leu11+ cells (B) of peripheral blood in patients with various liver diseases and ASCs were measured using double colour immunofluorescence assay of MoAb with FACS flow cytometry. 1) The mean % of IL2R+ Leu11+ cells which was 0.5 +/- 0.2 in healthy controls, decreased significantly in HCC in comparison with CALD, ASC and healthy controls, and in ASC rather than in healthy controls. They (A) were less than 0.1% in eight of thirteen cases with HCC, in one of twenty cases with CALD and of eleven ASCs, respectively. 2) In the mean % of HLADR+ Leu11+ cells which was 2.9 +/- 1.8 in healthy controls there was not a significant difference among HCC, CALD, ASC, AH and healthy controls. They (B) were less than 0.1% only in one case with HCC. 3) The value of fluorescence intensity of IL2R on IL2R+ Leu11+ cells reduced in B-CALD, ASC, AH and HCC, and one of HLADR on HLADR+ Leu11+ cells increased in CALD. These results suggested that the decrease of IL2R+ Leu11+ cells was due to the existence of HCC. PMID- 2560488 TI - [Serial change in serum concentration of type III procollagen N-terminal peptide after TAE in hepatocellular carcinoma, and analysis of gel filtration pattern of the peptide]. AB - In the present study, we have measured the serum concentration of PIIIP in patients with various liver diseases, and studied serial changes in serum PIIIP after TAE and its gel filtration pattern in 10 cases of hepatocellular carcinoma undergone TAE. The following results were obtained. 1) Serum concentration of PIIIP was 12.3 +/- 6.1 ng/ml in normal controls and elevated significantly in liver cirrhosis, liver cirrhosis with hepatocellular carcinoma, chronic active hepatitis, and acute hepatitis. 2) There was no significant difference in the serum concentrations of PIIIP between liver cirrhosis and liver cirrhosis with hepatocellular carcinoma. The result suggested that serum PIIIP cannot be a specific marker of hepatocellular carcinoma. However, the serum PIIIP concentration was decreased 2 or 4 weeks after TAE in effective cases, whereas increased in ineffective cases. Thus, the measurement of serial change in the serum PIIIP after TAE was considered to be useful for evaluating the effectiveness of TAE. 3) In analysing the elution patterns of serum PIIIP by gel chromatography, the peak of 125I-PIIIP antigen decreased 4 weeks after TAE in effective cases, whereas, no change was observed in the elution profile by gel chromatography 4 weeks after TAE in ineffective cases. These results seem to be caused by necrosis of hepatocellular carcinoma by TAE, and suggest the possibility that PIIIP is produced in hepatocellular carcinoma tissue. PMID- 2560489 TI - [Evaluation of the formation of esophageal varices by per-rectal portal scintigraphy]. AB - Portal circulation in patients with liver diseases was evaluated by 99mTc pertechnetate per-rectal portal scintigraphy, and we retrospectively examined the relationship between the extent of abnormality in the portal circulation and the development of esophageal varices. The per-rectal portal shunt index (PRPSI) was calculated for 13 healthy subjects and 79 patients with chronic hepatitis and 214 with cirrhosis of the liver. In the healthy subjects, the mean PRPSI was 4.8%. In the patients with hepatitis, the mean PRPSI was 8.4%, and in the patients with cirrhosis, it was 48.5%. The PRPSI was significantly higher in the cirrhotic patients with esophageal varices than in the without, and also in the cirrhotic patients with encephalopathy than in those without. The cumulative incidence of esophageal varices in the 3 years of the study in patients whose PRPSI was 20% or over was significantly higher than that in patients whose PRPSI was under 20%. The results suggested that this non-invasive method should be useful for predictions of the formation of esophageal varices. PMID- 2560490 TI - [A case of a collision tumor of hepatocellular carcinoma and gallbladder carcinoma, accompanied by anomalous pancreaticobiliary ductal union and congenital intrahepatic bile duct dilation]. PMID- 2560491 TI - [A case report of a vipoma of the pancreas which caused watery diarrhea and hypokalemia]. PMID- 2560493 TI - Studies on mechanisms of initiation and propagation of seizure activity following penicillin application. PMID- 2560492 TI - [A high risk group of hepatocellular carcinoma in man, with special reference to its clinical significance for the screening of early liver cancer. Gifu Study Group for Early Liver Cancer]. AB - A high risk group of hepatocellular carcinoma (HCC) was statistically established using multiple regression analysis of 331 cases with liver cirrhosis (LC), in which 78 cases later developed HCC. Highly contributing factors to hepatocarcinogenesis were found to be positive HBsAg, age, drinking history, sex (male), history of blood transfusion, history of acute hepatitis (or jaundice) and elevated plasma levels of alpha-fetoprotein. A prospective study was initiated in April, 1985 employing another 122 LC patients to clinically evaluate the significance of the high risk group of HCC. 28 cases with small HCC (less than 3 cm in diameter) were newly found: 4 with chronic hepatitis and 24 with LC, among whom 22 developed from the high risk group (sensitivity 92%, specificity 44%). Three year survival rate of 28 cases thus found was 56%, and causes of death of 9 fatal patients were 4 cancer death, 4 hepatic failure and 1 gastrointestinal bleeding. In conclusion, the high risk group is valuable for the screening of early liver cancers, and treatments of hepatic failure as well as of HCC itself are important to improve the prognosis of HCC patients thus diagnosed. PMID- 2560494 TI - Basic mechanism of epileptogenicity--consideration from studies of acute kindling. PMID- 2560495 TI - The seizure mechanisms of an epileptic mutant animal. PMID- 2560496 TI - Seizure-triggering mechanisms in the kindling model of epilepsy: collapse of GABA mediated inhibition and activation of NMDA receptors. PMID- 2560497 TI - Inositol trisphosphate (IP3) receptors and epileptic seizure. AB - The effects of anticonvulsants and Ca2+ channel antagonists on the inositol trisphosphate (IP3) binding and IP3-induced Ca2+ release were examined in brain membrane fractions. Anticonvulsant (PHT and valproate) and Ca2+ channel antagonists (verapamil, diltiazem, flunarizine, nicardipine and cinnarizine), examined did not significantly inhibit the IP3-receptor binding. The Kd and Bmax values of the IP3 binding did not change significantly in the various brain regions of the amygdala-kindled rats, killed 10 days after the last seizure, compared to those of controls. On the other hand, PHT, PB and carbamazepine inhibited the Ca2+ releasing activity of IP3 in the cerebellar membrane fractions by approximately 20% at therapeutic concentrations. PMID- 2560498 TI - Mechanism of phenytoin action at the cellular level. PMID- 2560499 TI - Neuroexcitotoxic action of db-cAMP: lesioning of neuronal cell bodies while sparing fibers of passage. PMID- 2560500 TI - Quantitative autoradiographic study of benzodiazepine receptors: comparative study between normal ddY mouse and El mouse. PMID- 2560501 TI - Long-term enhancement of excitatory amino acid receptor-mediated polyphosphoinositide hydrolysis in the hippocampus and amygdala/pyriform cortex of hippocampal kindled rats. PMID- 2560502 TI - [A case of hypersensitivity pneumonitis due to isocyanate (TDI)]. AB - A 46-year-old man who had worked in a paint processing plast for over 29 years was admitted to our hospital with complaints of nocturnal dyspnea and dry cough. A chest X-ray film showed diffuse granular shadows in bilateral lungs. Pulmonary function tests revealed reduction of diffusing capacity and restrictive impairments. Hypersensitivity pneumonitis (HP) due to isocyanates was speculated from his occupational history and clinical course. Positive skin tests against TDI-HSA and MDI-HSA, precipitating antibody against TDI-HSA, and negative lymphocyte stimulating tests of peripheral blood and bronchoalveolar lavage fluid were also noticed. Environmental provocation test was positive. Histological findings of transbronchial lung biopsy specimens showed diffuse alveolitis and Masson body, but no granulomas. According to these results, the patient was diagnosed as HP due to TDI. Type III allergy of Gell-Coombs seems to participate in this case. The granulomatous lesion is seen less frequently in isocyanate related HP than in HP induced by organic dusts, which suggests the difference in immunological and histological reactions between both types of HP. PMID- 2560503 TI - [Effect of fish oil diet on activities of lipogenic enzymes and glucose-6 phosphatase in rat liver and adipose tissue]. AB - In order to evaluate the effect of fish oil on lipid hydrogenase(G6PDH), malic enzyme(ME), glucose-6-phosphatase(G6Pase) activities were measured in liver and adipose tissue of rats fed 13 days supplemented fish oil at the level of 10% (W/W). Two other groups of rats were fed 10% soybean oil or lard to compare with the effect of fish oil. In all groups, activities of hepatic G6PDH and ME were depressed from the beginning of feeding. This effect was greatest (50%) in fish oil group. Hepatic G6Pase was highest in rats fed lard. When the level of fish oil was reduced to half, as total fat content was maintained at the level of 10% by complementary lard, lipogenic enzyme depressing effect of fish oil was as significant as shown in 10% fish oil diet. Hepatic G6PDH was depressed significantly (14%) in rats fed fish oil as low as 2%. On the other hand, changes in adipose tissue G6PDH and ME activities were small. Adipose tissue G6PDH activity increased slightly in rats fed with increasing fish oil(above 0.5%). It is suggested that fish oil alter, more markedly than either soybean oil or lard, cellular lipid metabolism by reducing activities of hepatic lipogenic enzymes. PMID- 2560504 TI - [Reoperation of the liver]. PMID- 2560505 TI - [The effect of chronic kidney failure on electroneurographic changes in patients on hemodialysis treatment]. AB - Electroneurographic examination of n. medianus, n. ulnaris, n. tibialis and n. fibularis was performed in 23 patients (15 men and 8 women) at mean age 52.1 years and mean duration of hemodialysis treatment 42.1 months. All patients had electroneurographic symptoms of peripheral polyneuropathy. The duration of hemodialysis treatment had no effect on the severity of the neurologic injury. Significant correlation was found between the severity of anemia and the peripheral nerve injury. According to the authors' data, essential for the development and severity of the uremic polyneuropathy was also the severity of the uremic syndrome. PMID- 2560506 TI - [A case of small cell carcinoma of the lung with marked hemorrhagic syndrome]. PMID- 2560507 TI - [Regulation of fibrinolysis by lymphocytes in inflammatory diseases of the bronchopulmonary system]. AB - Urokinase receptors on lymphocytes described in the paper link immune system of the body with fibrinolytic one, play a biological role in sano-++- and pathogenesis in aggravation of chronic purulent bronchitis, of focal and croupous pneumonia. A correlation between the trend in the expression of urokinase receptors on lymphocytes and changes in activity of plasminogen activator in the blood and urine was established: the more the expression, the more the activity. The method can serve a criterion of the treatment effectiveness+ in nonspecific inflammatory bronchial and pulmonary diseases and be helpful in designing methods of their treatment improvement. PMID- 2560508 TI - [Cancer of the liver (analysis of autopsy data)]. AB - The findings at 26.879 autopsies, that is 70% of all the deaths registered in the Province of Trieste, obtained at the Institute of Pathologic Anatomy and Histology of Trieste from 1976 to 1985 were studied. Liver cancer showed in the Province the World Standardized Incidence Rate (WR) of 1.87 (females) and 12.78 (males), one of the highest ever observed in nonendemic areas. In view of close association between cancer of the liver and cirrhosis (88%), the authors studied the most important etiological factors of cirrhosis in the region, i.e. hepatitis B and alcohol abuse. Some effective measures are suggested to decrease the incidence of cirrhosis and eventually cancer of the liver. PMID- 2560509 TI - Binding of steroid receptors to the HREs of mouse mammary tumor virus, chicken and xenopus vitellogenin and rabbit uteroglobin genes: correlation with induction. AB - Binding to hormone responsive elements (HRE) is an essential step in gene regulation by steroid hormones. Using a combination of in vivo and in vitro studies we have analyzed the interactions of the estrogen receptor with genes from three different systems: the long terminal repeat (LTR) of the mouse mammary tumor virus (MMTV), the chicken and xenopus vitellogenin genes and the rabbit uteroglobin gene. The estrogen receptor binds to all four genes in vitro, but the MMTV LTR does not respond to estrogen in gene transfer experiments. Similarly, the xenopus vitellogenin gene binds the progesterone and glucocorticoid receptors in vitro, but only estrogen induces the xenopus vitellogenin gene in gene transfer. These results suggest that events distal to DNA binding are essential for transcriptional activation, and that the function of the HRE is not simply to position the hormone receptor in the vicinity of the regulated promoter. PMID- 2560510 TI - The regulation of expression of mouse mammary tumor virus DNA by steroid hormones and growth factors. AB - Mouse mammary tumor virus (MMTV) expression is associated with hyperplastic alveolar growth and subsequent development of mammary cancers in the mouse. The expression of this virus is also controlled by factors involved in the normal proliferation and differentiation of the mammary epithelium. During pregnancy when the mammary gland undergoes massive proliferation, MMTV expression is increased. Steroid hormones and growth factors that play an important role in the proliferation of mammary gland cells are responsible for the increased MMTV expression. In sarcomatous transformation of mouse mammary epithelial cells, MMTV expression is repressed. This repression is due to negative control of MMTV expression by transforming growth factor-beta (TGF beta). This growth factor is produced in high amounts when mammary epithelial cells progress into the transformed state. The expression of MMTV is therefore under multiple control by steroid hormones and growth factors. PMID- 2560511 TI - Importance of estrogen sulfates in breast cancer. AB - Estrogen sulfates are quantitatively the most important form of circulating estrogens during the menstrual cycle and in the post-menopausal period. Huge quantities of estrone sulfate and estradiol sulfate are found in the breast tissues of patients with mammary carcinoma. It has been demonstrated that different estrogen-3-sulfates (estrone-3-sulfate, estradiol-3-sulfate, estriol-3 sulfate) can provoke important biological responses in different mammary cancer cell lines: there is a significant increase in progesterone receptor. On the other hand, no significant effect was observed with estrogen-17-sulfates. The reason for the biological response of estrogen-3-sulfates is that these sulfates are hydrolyzed, and no sulfatase activity for C17-sulfates is present in these cell lines. [3H]Estrone sulfate is converted in a very high percentage to estradiol (E2) in different hormone-dependent mammary cancer cell lines (MCF-7, R 27, T-47D), but very little or no conversion was found in the hormone-independent mammary cancer cell lines (MDA-MB-231, MDA-MB-436). Different anti-estrogens (tamoxifen and derivatives) and another potent anti-estrogen: ICI 164,384, decrease the concentration of estradiol very significantly after incubation of estrone sulfate with the different hormone-dependent mammary cancer cell lines. No significant effect was observed for the uptake and conversion of estrone sulfate in the hormone-independent mammary cancer cell lines. Progesterone provokes an important decrease in the uptake and in estradiol levels after incubation of [3H]estrone sulfate with the MCF-7 cells. It is concluded that in breast cancer: (1) Estrogen sulfates can play an important role in the biological response of estrogens; (2) Anti-estrogens and progesterone significantly decrease the uptake and estradiol levels in hormone-dependent mammary cancer cell lines; (3) The control of the sulfatase and 17 beta-hydroxysteroid dehydrogenase activities, which are key steps in the formation of estradiol in the breast, can open new possibilities in the treatment of hormone-dependent mammary cancer. PMID- 2560512 TI - Regulation of SBP synthesis in human cancer cell lines by steroid and thyroid hormones. AB - The presence of human sex steroid binding protein (SBP) in liver cells, the supposed site of SBP synthesis, and in other target cells for sex steroid hormones such as breast, endometrium and prostate epithelium, have been demonstrated by indirect immunofluorescence. It is not known whether SBP enters endometrial and prostate cells by endocytosis, possibly mediated by a cell membrane receptor process, or if SBP is synthesized in these cells. SBP mRNA has been searched in human cancer cell lines originated from liver (Hep G2/H5A), breast (MCF-7), endometrium (RL95-2) and prostate (LNCaP). It was only found in hepatoma cells where it is regulated by estradiol, antiestrogen tamoxifen and triiodothyronine, in a similar way as secreted SBP. This work provides evidence that human SBP is synthesized in the liver, and it also suggests that its regulation may involve non-steroidal factors. PMID- 2560513 TI - Mechanism of virus-induced stimulation of the hypothalamus-pituitary-adrenal axis. AB - Increased blood levels of glucocorticoids are observed during certain viral infections. In this paper, we report data obtained from a model in rodents showing that the pituitary-adrenal axis is stimulated following inoculation of Newcastle Disease Virus (NDV). No evidence for an ectopic, lymphoid source of ACTH-like immunoreactive material capable of inducing this effect was obtained. Administration of virus-free supernatants from cocultures of human peripheral blood leukocytes with NDV also stimulated ACTH and glucocorticoid output in normal mice. This observation showed the immunological cell origin of the mediator of the hormonal effect. Pretreatment of the supernatant with anti-IL-1 sera neutralized its capacity to induce an increase in glucocorticoid and ACTH levels in blood. Furthermore, injection of IL-1 in nanogram amounts also increased ACTH and glucocorticoid blood levels. Thus, we conclude that IL-1 is the most likely mediator of the stimulation of the pituitary-adrenal axis during viral infection. The reported data are also discussed in the general context of the postulated glucocorticoid-associated immunoregulatory circuit. PMID- 2560514 TI - The effects of 9-ene-tetrahydrocannabinol on hormone release and immune function. AB - We investigated effects of 9-ene-tetrahydrocannabinol (THC) on endocrine and immunological function. Seventeen male volunteers entered into a double blind, randomized study to receive oral THC (10 mg t.i.d. for 3 days and on the morning of the fourth day) or placebo, after at least 2 weeks of abstinence. Plasma prolactin, ACTH, cortisol, luteinizing hormone and testosterone were not altered during or after THC, compared with baseline concentrations. Tests of lymphocyte function showed no differences compared to baseline between THC and placebo groups. As the relatively low dosing regimen of THC (10 mg t.i.d.) resulted in no alterations, another group of 6 men were administered higher doses of THC by inhalation (18 mg/marijuana cigarette) following the same dosing regimen. No endocrine or immunological alterations were observed. When the subjects were grouped according to their history of THC use prior to admission, heavy THC users had lower prolactin concentrations than light users. No differences were observed in concentrations of other hormones or in tests of immune function. PMID- 2560515 TI - Adaptation of aldosterone biosynthesis to sodium and potassium intake in the rat. AB - The steroidogenic response of rat adrenal zona glomerulosa to stimulators is variable and depends on the activity of biosynthetic steps involved in the conversion of deoxycorticosterone (DOC) to aldosterone (Aldo). Corticosterone methyl oxidations (CMO) 1 and 2 are stimulated by sodium restriction and suppressed by potassium restriction. These slow alterations are accompanied by the appearance or disappearance of a specific zona glomerulosa mitochondrial protein with a molecular weight of 49,000. Induction of CMO 1 and 2 activities and the appearance of the 49 K protein can also be elicited in vitro by culture of rat zone glomerulosa cells in a medium with a high potassium concentration. The 49 K protein crossreacts with a monoclonal antibody raised against purified bovine adrenal cytochrome P-450(11 beta). The same antibody stains a protein with a molecular weight of 51,000 in rat zona fasciculata mitochondria and in zone glomerulosa mitochondria of rats in which CMO 1 and 2 activities have been suppressed by potassium restriction and sodium loading. The 51 K crossreactive protein was purified to electrophoretic homogeneity by chromatography on octyl sepharose. In a reconstituted enzyme system, it converted DOC to corticosterone (B) and to 18-hydroxy-11-deoxycorticosterone (18-OH-DOC) but not to 18 hydroxycorticosterone (18-OH-B) or Aldo. A partially purified 49 K protein preparation from zona glomerulosa mitochondria of rats kept on a low-sodium, high potassium regimen converted DOC to B, 18-OH-DOC, 18-OH-B and Aldo. According to these results, rat adrenal cytochrome P-450(11 beta) exists in two different forms, with both of them capable of hydroxylating DOC in either the 11 beta- of the 18-position, but with only the 49 K form capable of catalyzing CMO 1 and 2. The adaptation of aldosterone biosynthesis to sodium deficiency or potassium intake in rats is due to the appearance of the 49 K form of the enzyme in zona glomerulosa mitochondria. PMID- 2560516 TI - Insulin enhances glucocorticoid induction of gene expression in a sequence specific manner. AB - Glucocorticoid receptor (GR) induces transcriptional activation of specific genes by binding to cis-acting enhancer sequences termed glucocorticoid responsive elements (GREs). Insulin regulates gene transcription part of which is mediated sequences dominated by GREs. By transient expression in HTC cells of fusion gene consist MMTV 5' flanking sequences containing the GREs on the thymidine kinase promoter (TK) and the coding region of the chloramphenicol acetyl transferase gene (CAT), we demonstrate that insulin increases 2-fold the produced induction from dexamethasone, while insulin alone does not exceed any significant effect. This synergistic effect of insulin is sequence specific since, expression of the glucocorticoid unresponsive gene RSV-CAT at the same cell line is affected neither by insulin nor dexamethasone or both. Transient expression of CAT constructs with progressive 5' deletion mutants of mouse mammary tumor virus (MMTV) promoter, fail to detect a cis-acting insulin responsive DNA element, that participates in increased dexamethasone mediated induction. We believe that insulin activates an intracellular mediator-transcription factor, that stimulates gene expression indirectly, by interaction with the glucocorticoid receptor. PMID- 2560517 TI - Hydrophobic cluster analysis (HCA) of the hormone-binding domain of receptor proteins. AB - A new technique of protein sequence analysis, namely, Hydrophobic Cluster Analysis (HCA), has been used to align and compare the sequences of proteins belonging to the receptor superfamily (steroid, thyroid hormone and retinoic acid receptors) and serpin superfamily (corticosteroid binding globulin (CBG) and alpha 1-antitrypsin (alpha 1-AT]. By matching up clusters of hydrophobic amino acids that oftenmost correspond to identifiable secondary structures (alpha helices, beta-strands etc.), it has been possible to deduce the following information on the secondary structures of these proteins: CBG is structurally related to alpha 1-AT (HCA score greater than 80%), the structures of the hormone binding domains of the steroid receptors that bind 3-keto-delta 4-steroids are closely interrelated (greater than 80%) but less closely related to that of the estrogen receptor (ER) (approximately 75%), vitamin D, retinoic acid and thyroid hormone receptors are structurally closely related (greater than or equal to 80%). Their secondary structures are, however, also related to that of the steroid receptors (approximately 70%), and a high degree of analogy exists between the structures of serpins and of the hormone-binding domains of members of the steroid superfamily (60-70%). HCA has clearly shown that a previous local sequence alignment of the estrogen receptor with other steroid receptors and cytochromes P450 has to be reconsidered. The published consensus steroid binding sequence previously identified in cytochromes is in fact 80 amino-acids upstream from its previously defined position. Other regions of contiguous sequence identity have also been identified which may be involved in the hydrophobic core of the protein or in steroid binding. Their positions have been indicated using the crystal structure of alpha 1-AT as a model. PMID- 2560519 TI - Hormonal regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase mRNA in the rat adrenal gland. AB - We studied the effect of ACTH on 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase enzyme. Reductase activity and reductase mass were enhanced by 22- and 6.2-fold respectively in one series of experiments, whereas in another the levels of reductase activity, reductase mass, and reductase mRNA were increased 6.6-, 3.6- and 2.2-fold respectively, following daily administration of exogenous ACTH for 3 days. Daily injection of 4-aminopyrazolopyrimidine (4-APP) to rats for 3 days increased circulating ACTH level 5.4-fold, whereas adrenal HMG-CoA reductase activity, reductase mass and reductase mRNA levels were greatly increased 36-, 10 and 16-fold, respectively. To counteract the effect of elevated plasma ACTH, dexamethasone acetate (Dex) was administered to 4-APP treated rats. At 3 h post Dex administration, plasma ACTH and corticosteroids levels were effectively decreased by 58 and 59%, respectively. The levels of adrenal HMG-CoA reductase mRNA, reductase activity and reductase mass were also diminished by 38, 31 and 40%, respectively. Our results show that rat adrenal HMG-CoA reductase can respond rapidly to hormonal changes, presumably through variations in circulating ACTH levels. PMID- 2560518 TI - Ligand-dependent interaction of the dioxin receptor with target DNA. AB - Wild type and nuclear transfer deficient mouse hepatoma cell lines were used to study the specific DNA binding of a dioxin inducible factor. This factor interacts with XRE only after dioxin treatment and is absent in receptor mutant containing cells even after treatment. Thus, evidence is provided to substantiate the claim that the dioxin receptor is involved in the specific DNA interaction with dioxin response enhancer elements. It is also shown that the molybdate stabilised dioxin-receptor interacts with hsp90 suggesting that, in similarity to the glucocorticoid receptor, the dioxin receptor is kept in a non-transformed state in the absence of ligand. PMID- 2560520 TI - New analogues of mifepristone with more dissociated antiprogesterone activities. AB - Mifepristone (RU 486 or RU 38486) possesses strong antiprogesterone and antiglucocorticoid along with moderate antiandrogen properties, which would limit its use in some therapeutic applications. In a search for more dissociated derivatives, the hydroxy substituent and the propynyl group in position 17 of the RU 486 series was replaced by a spiroether group, which is known to induce specific affinity for the progestin receptor in steroid series. The substituents in the para position of the 11 beta-phenyl group, leading to the most potent derivatives in the RU 486 series, were retained. The new derivatives have been studied in vitro for their relative binding affinities (RBAs) for the steroid receptor and in vivo for their hormonal and antihormonal activities. The selected compounds, RU 46556 and RU 49295 display the following properties: in vitro, like RU 486, they show a strong RBA for the rabbit progestin receptor, but a much lower one for the rat thymus glucocorticoid receptor; in vivo they are about three times more active than RU 486 for inducing abortion in rats, but unlike the latter they are devoid of any antiglucocorticoid activity on the thymus weight in rats. These antiprogesterone effects have been confirmed on the deciduoma formation in rats and on the endometrial proliferation in rabbits. However, in contrast to RU 486 in the latter test, some progestomimetic activity has been observed. RU 46556 and RU 49295 are now under extensive pharmacological study. PMID- 2560521 TI - In vivo effects by estrone sulfate on the central nervous system-senile dementia (Alzheimer's type). AB - Seven women with senile dementia-Alzheimer's type (SDAT) were treated with conjugated estrogen [main content: estrone sulfate (E1-S)], at a dose of 1.25 mg/day over a 6-week period. A New Screening Test for Dementia developed by Japanese National Institute of Mental Health (NS) and the scores of Hasegawa Scale for dementia (HS) were performed every 3 weeks. Six women showed improvements in NS (P less than 0.05) and 5 women showed improvements in HS. Untreated women with SDAT did not show any improvement. Serum E1-S was measured by a direct radioimmunoassay. Serum E1-S was 911 +/- 156 pg/ml in 7 women with SDAT and lower than that of 7 normal women (1020 +/- 216 pg/ml). Following the treatment, serum E1-S increased to a level of 21.1 +/- 8.1 ng/ml. Estrone and estradiol-17 beta also increased. The results suggest a possibility for the future clinical use of estrogen for senile dementia, after careful clinical research trials including the side effects. PMID- 2560522 TI - Effects of aldosterone on (Na+ + K+)-ATPase of amphibian sodium-transporting epithelial cells (A6) in culture. AB - Stimulation by aldosterone of sodium reabsorption can be reproduced on a cell line, A6, derived from the renal tissue of Xenopus laevis. These cells organize themselves as a polarized epithelium carrying out unidirectional sodium transport, reflected by the short-circuit current (Isc). Isc response to aldosterone starts to be apparent after a latency period of 2-3 h; the full hormonal effect takes much longer. On the other hand, (Na+ + K+)-ATPase activity and density in ouabain binding sites did not increase before several hours of treatment. At that stage, while Isc more than trebled, Na+ pump activity and density went up by less than 50%. A significant influence of aldosterone on the way the Na+ pump operates is considered unlikely, since cell interaction with ouabain remained unchanged (Kd approximately 18 nM). Furthermore, the close correspondence of hormonal effect, in relative terms, on (Na+ + K+)-ATPase activity vs density, argues against a significant degree of recruitment of spare pump units. Thus aldosterone effect on Na+ pump probably results from increased biosynthesis of the enzyme. The aldosterone dependent Na+ pump stimulation is apparently unrelated to sodium available for transport. The hormone seems to act on Na+ pump directly. PMID- 2560523 TI - [Angiotensin converting enzyme in the blood and lungs in experimental diabetes]. AB - The activity of angiotensin converting enzyme has been examined in serum and lungs of Wistar male rats with streptozotocin induced diabetes. The diabetes has been induced by single i.v. dose of streptozotocin (75 mg/kg body weight). The activity of angiotensin converting enzyme in serum and lungs has been measured ten days after application of drug by spectrophotometrical method using synthetic substrate hippuryl-1-histidyl-1-leucine. The results show that streptozotocin induced diabetes in rats cause significant increase in angiotensin converting enzyme activity in serum and lungs. PMID- 2560524 TI - State of hepatitis B viral genomes in cirrhotic and hepatocellular carcinoma nodules. AB - DNA of individual cirrhotic nodules (CN) and hepatocellular carcinoma nodules (HCN) of three hepatitis B surface antigen positive autopsy cases with macronodular cirrhosis were analyzed by Southern blot and slot blot hybridization with a hepatitis B virus (HBV) DNA probe. Evidence of episomal or replicating viral DNA, viral DNA integration at the same cellular DNA site in many cells (clonal integration) and viral integration in different cellular DNA sites in many different cells (non-clonal integration) was found in different cirrhotic nodules of the same liver, indicating heterogeneity in the state of HBV in different cells and in different cirrhotic nodules within each infected liver. Episomal or replicating viral DNA forms were found in all cirrhotic nodules of one liver, in less than 10% of examined nodules of a second liver and in none of the third. Evidence of clonal viral integration was found in CN of all three livers and non-clonal integration in CN of the latter two. Cirrhotic nodules with apparent different integrations in many different cells (non-clonal integration) outnumbered those with the same integration site in many cells (clonal integration), and many cirrhotic nodules in those two livers had no detectable viral DNA. Cirrhotic nodules with a viral integration in the same cellular DNA site in many cells would appear to have been formed by clonal expansion of an original cell containing the viral integration, and cirrhotic nodules with different integrations in many different cells (non-clonal integration) may have been formed by recruitment of many different cells with different viral integrations or by clonal expansion of cells without HBV integrations and subsequent viral integrations occurring integration. In one liver, three different hepatocellular carcinoma nodules appeared to represent metastatic lesions because the clonal pattern of HBV integration was identical in each, and in another liver different HCN appeared to be of different clonal origin, i.e. to have arisen from different cells, because multiple viral integrations (i.e. multiple individual restriction fragments with HBV sequences) were each different in different HCN of that liver. PMID- 2560525 TI - Congenital varicella-zoster virus infection and the failure to establish virus specific cell-mediated immunity. AB - Varicella-zoster virus (VZV) is one of the six human herpesviruses. The viral genome encodes five glycoproteins among its 70 open reading frames; these have been designated gpI to gpV. VZV causes the primary disease chickenpox, usually in children, after which the virus remains latent in the dorsal root ganglia. Later in life, VZV reactivates and causes the disease zoster. VZV can also infect the fetus of a pregnant woman who contracts chickenpox. The fetopathy is unusual in that it more closely resembles zoster than chickenpox. To determine whether the intrauterine immune response is impaired following VZV infection, the humoral and cellular immune responses were first defined in healthy children and adults following chickenpox. All produced virus-specific antibody responses to the viral glycoproteins; in addition, their lymphocytes proliferated when stimulated by both crude VZV antigen and purified glycoprotein products. The fetal immune system generated immunoglobulin M-specific antibodies to the individual VZV glycoproteins. However, no lymphocyte proliferative response was detected. Thus, these studies suggest that the fetus may not be able to mount a cell-mediated response to VZV antigens and that this impaired immunity may contribute to the severe sequelae. PMID- 2560526 TI - [The role of viral antibodies in the pathogenesis of degenerative and demyelinating diseases]. AB - Serologic investigations were carried on samples of sera and cerebrospinal fluid from patients aged between 6 and 67 years with degenerative and demyelinating disease with the aim of determining the pathogenetic role of viruses in these disorders. Increased titers of complement fixation antibodies against the measles virus were detected in patients with subacute sclerosing panencephalitis and multiple sclerosis, whereas high titers of complement fixation antibodies against Herpes Simplex Virus Type I were found in Parkinsonian cases. Demonstration of high titers of heterophil antibodies in patients with Parkinson's disease seemed to be a significant contribution to this matter. Although HSV type I and heterophil antibodies were high in patients with amyotrophic lateral sclerosis, small number of samples hampered satisfactory interpretation. PMID- 2560527 TI - [The viral etiology of amyotrophic lateral sclerosis]. AB - Roles of immunologic factors and slow virus infections in the aetiopathogenesis of amyotrophic lateral sclerosis (ALS) have recently gained significance. For this purpose, presence of poliovirus type I, measles and herpes simplex (HSV) type I antibodies were determined in the sera of ALS patients by complement fixation technique. Significant increase in antibody titers for poliovirus type I and HSV type I were determined in sera of ALS patients in comparison with controls and patients with other neurologic disorders. PMID- 2560528 TI - [Human B-lymphotropic virus (HBLV): human herpesvirus-6 (HHV-6)]. AB - Human B-lymphotropic virus (HBLV) is also known as human herpes virus-6 (HHV-6) and was isolated from patients with various lymphoproliferative disorders. In this article the biological, immunological and molecular properties of a new human herpes virus-6 is reviewed. The association of the virus with chronical disorders is also discussed. PMID- 2560529 TI - [Immunohistochemical study of cancer-associated carbohydrate antigens in carcinoma of the biliary tract]. AB - To reveal the cell-biological character of biliary tract cancer, localization and distribution of three cancer-associated carbohydrate antigens (CA19-9, sialyl SSEA-1, NCC-ST-439) and carcinoembryonic antigen (CEA) were studied immunohistochemically in 35 cases of gallbladder carcinoma, 21 of bile duct carcinoma, 16 of chronic cholecystitis, and 3 of normal gallbladder. 1) All carbohydrate antigens and CEA were present in 70-90% of the cases of gallbladder and bile duct carcinoma. In particular, NCC-ST-439 had the highest incidence of positive staining (95.2%) in bile duct carcinoma. 2) The mode of localization was diverse and was not fixed by the kind of antigen. Antigens flowing out to the surrounding stroma were affected by the rate of positive cells. 3) No significant correlation was observed between the histological type or degree of differentiation and tissue positivity. 4) The positivity of tissue CEA was higher in the cases with serous membrane invasion, gamma INF pattern, and neuro-, vascular-, and lymphatic invasion. 5) In chronic cholecystitis, CA19-9, NCC-ST 439, and CEA were stained in mucosal cells and/or metaplastic cells, while sialyl SSEA-1 was stained only in one case in the goblet cells and the cells with pseudopyloric metaplasia. None of the antigens were stained in normal gallbladders. These results suggest that these antigens may be useful in the diagnosis and therapeutic treatments in patients with biliary tract cancer. PMID- 2560531 TI - Ultrastructure of the sarcomatous component in gliosarcomas. PMID- 2560530 TI - On the specificity of focal thickenings of myelin in peripheral nerves. PMID- 2560533 TI - Epstein-Barr virus associated lymphoproliferative diseases (B cell lymphoma) after transplantation. AB - We report 12 cases of lymphomas which occurred among 1670 patients with kidney or combined renal and pancreatic transplantation. Group 1 comprised nine patients presenting with the diffuse form of the disease where immunoblasts or mature plasma cells massively infiltrated all organs. The first symptom was a viral syndrome, associated with a restriction of heterogeneity of immunoglobulins; oligoclonal to monoclonal peaks of immunoglobulins appeared about 50 days after transplantation. All patients received antilymphocyte globulins (ALG), and seven were treated with cyclosporin. EBV infection could be demonstrated in almost all patients; three EBV lymphoblastoid cell lines were established, their HLA phenotype being the same as the recipient of the graft. All patients finally died with renal and hepatic failure. Group 2 comprises three patients who presented solid B cell tumours of tonsils, lungs, and spleen at onset, extending to liver, kidney graft, lymph nodes, and brain. All received cyclosporin; two patients were treated with ALG, and one with OKT3. Immunoglobulins were polyclonal, oligoclonal, or decreased. Cell surface immunoglobulins were monoclonal on two tumours. EBV-DNA was positive within two tumours. Two patients presented EBV and CMV primary infection. CD4+T lymphocytes subsets were diminished at onset, and increased after cessation of immunosuppressive therapy. One patient died because of brain involvement; the two others are alive, one with perfect graft function. Therapy consisted of stopping immunosuppressive treatment, Acyclovir, and in two patients of group 2, monoclonal antibodies to pan-B and EBV receptor antigens. PMID- 2560532 TI - [Gigantic fibroadenoma of the breast in an adolescent. Description of a clinical case]. AB - A case of giant fibroadenoma in a girl of 14 is reported. This case emerged from an examination of about 380 patients observed at the Senology Clinic of the S. Maria di Loreto Hospital (USL 44) in the last 3 years. A statistical contribution is made to the study of breast pathologies in pubertal age and stress is laid on the importance of mass screening among adolescents. PMID- 2560534 TI - Diazepam effects on the fetus. AB - Treatment of time-pregnant Long Evans rats with 1.25 mg/kg s.c. diazepam (2.5 mg/kg in Sprague Dawley rats) from gestational day 14 to 20 produced transient depression of an olfactory guided behavior (nest odor behavior) in suckling offspring. Enhanced drug sensitivity to diazepam was seen in adult male and female off-spring as indicated by increased temperature depression. In addition, increased sensitivity to an opiate (morphine) was noted for the female offspring in the tail flick test. Treatment of the pregnant dam with diazepam or clonazepam, a benzodiazepine with selective affinity for the central benzodiazepine receptor, resulted in a marked depression of cellular immune responses in the offspring of both sexes up to 2 months of age. Drug treatment during early fetal period (GD 12-16), at a time central benzodiazepine receptors are not present in all brain regions of the fetal brain, did not affect the quality of cellular immune responses, whereas treatment from GD 16 to 20 was effective. Prenatal diazepam effects are discussed in view of presence and functionality of both central and peripheral benzodiazepine binding sites in the fetus. PMID- 2560535 TI - Prenatal lorazepam administration is associated with GABAA receptor alterations in late embryonic and mature chicks. AB - Prenatal benzodiazepine administration has been associated with alterations in behavior in young and mature animals. Prior studies of neurochemical changes in animals treated in utero have produced conflicting results. We used a chick embryo system to assess the effects of chronic lorazepam administration on binding and function at the GABAA receptor in late embryos and in mature animals. Administration of lorazepam for 10 days of embryonic development (E8-E18) led to decreases in binding of the chloride channel ligand TBPS and in muscimol stimulated chloride uptake in late embryos (E18). Similar alterations were observed in mature animals after the same regimen of prenatal lorazepam. Persistent alterations in GABAA receptor binding and function after prenatal lorazepam may be due to effects of neuronal differentiation or on receptor regulation. These neurochemical effects may underlie the behavioral abnormalities associated with prenatal benzodiazepine administration. PMID- 2560536 TI - The effect of passive immunization on active immunity against Clostridium perfringens type D in lambs. AB - Lambs in different stages of development of active immunity against Clostridium perfringens type D were treated with partially purified immunoglobulin in an attempt to superimpose a passive immunity on an existing or developing active immunity. Three different studies were undertaken to determine the impact of partial purified immunoglobulins on these vaccinated animals. In 2 of the 3 studies, active immunity was induced by administering the normal routine enterotoxaemia vaccinations and allowing the basic immunity to become established, for a period ranging from 2 weeks for the animals in study 1 and 4 months for those in study 2, before passive immunization with the partially purified immunoglobulins took place. An increase in the epsilon antibody titre occurred in each of the 2 studies after the animals were passively immunized with immunoglobulin, though this increase was not statistically significant (P greater than 0.05). In the 3rd study, when the animals were given the initial vaccination of the Onderstepoort enterotoxaemia oil adjuvant vaccine together with the immunoglobulin, an immediate increase in the epsilon antitoxin titre occurred that was statistically significant (P less than 0.05) 2-14 days after administration. No negative effects were noted on the development of an initial active immunity or an existing active immunity against Clostridium perfringens type D when they were passively immunized with partially purified immunoglobulin. PMID- 2560537 TI - Prognostic parameters in breast cancer. AB - Carcinoma of the breast is the most common cancer in Australian women. Current methods of treatment and refinements in therapeutic regimens are based on our understanding of the biological behaviour of the disease. Several prognostic parameters have been identified which predict survival and allow the selection of patients who may benefit from adjuvant therapies. The more important of these parameters include tumour size, histologic type, histologic grade, axillary lymph node status, estrogen receptor status and tumour growth fraction. Diagnostic pathologists who are responsible for the evaluation of these parameters should be cognizant of their relative prognostic values and also of other factors such as cellular antigens, lectin binding and oncogenes which may have potential roles in predicting survival and therapeutic responses. This review provides an update of prognostic parameters which are assessed through examination of the excised specimen. PMID- 2560538 TI - An epidemic of adenovirus type 1 conjunctivitis. PMID- 2560539 TI - Freeze-fracture morphometry on normal and cancer breast tissues. AB - A stereological procedure to evaluate the number of particles per unit surface area (Np/micron2) and the particle density distribution in both protoplasmic (pf) and external face (ef) of the plasma membrane was applied on freeze-fracture preparations of human breast tissues with invasive carcinoma and with non neoplastic lesions. A significant higher intramembrane particle density was recorded on the protoplasmic face of plasma membrane of epithelial cells from non neoplastic breast tissues (2999 +/- 720 Np/micron2) compared to the particle density on the corresponding face of cancer cells (911 +/- 608 Np/micron2). Moreover, in the latter cells the external face had more than 50% of the surface area totally smooth while residual particles were often aggregated. The reduction of integral glycoproteins in the plasma membrane of malignant tissues fits apparently well with the increased activity of the methyltransferase-I, an enzyme methylating membrane phospholipids. These findings suggest a distribution of constituents and a lipid environment peculiar for the plasma membrane of breast cancer cells. PMID- 2560540 TI - Quantitative aspects in defining prognostic factors of breast cancer. AB - Although many attempts have been made to define prognostic criteria, the long term prediction of breast cancer prognosis remains a difficult task. In search of morphologic criteria with prognostic significance, we investigated 101 ductal invasive breast carcinomas by means of computerized image analysis. Our purpose was to define prognostic measures of the basic characteristics of breast cancers, growth rate and metastatic potential. Consequently, three principal types of analysis were performed: on Feulgen stained nuclei, on histological structures by using differential stainings and on desmosome-complexes revealed by immunofluorescence microscopy. The data obtained from analysis of the cell nuclei morphology and the chromatin structure served to establish objective nuclear grading. In comparison to the visual grading made during routine histology, the objective grading correlates better with the existence of lymph node metastases and with receptor status. The description of desmosome-complexes by using quantitative immunofluorescence microscopy indicates that in order to obtain relevant results, the histological pattern of the tumors has to be considered. In order to characterize these structures we introduced special methods of cluster analysis. The studies are an example of complex quantitative analysis of tumor morphology performed with different combined methods of microscopy and image processing. PMID- 2560541 TI - Flow cytometric analysis of DNA content in cervical lesions. AB - Flow cytometry is used to screen gynaecologic specimens for Cervical Intraepithelial Neoplasia (CIN) and its precancerous lesions, possibly associated with human papilloma virus (HPV), in order to investigate the role of aneuploidy as a biological marker in HPV and CIN lesions. A total of 299 cervical samples was collected by scraping and the cellular DNA content was measured using the propidium iodide staining procedure. Two groups of patients were studied; a group of 142 negative controls for cytology and groups of patients assigned to mild, moderate or severe dysplasia, carcinoma in situ diagnosed by histologic classification according to the Papanicolaou staining technique. Pathological samples showing an alteration of the DNA index or perturbation of the cellular phase S compartment ranged from 6.4% to 41.9%. Our results confirm those obtained by other authors and suggest the hypothesis that aneuploidy is present with permanence of viral DNA in early stages of carcinogenesis, which can be used as a marker in the transition from benign to malignant cells. This work is of potential interest for the possible follow-up of patients having condyloma and could aid the early diagnosis of cervical carcinoma. PMID- 2560542 TI - Grading and prognosis of invasive ductal mammary carcinoma by nuclear image analysis in tissue sections. AB - In primary tumours of 84 patients with invasive ductal carcinomas the prognostic significance of the nuclear image was assessed. Feulgen stained tissue sections were investigated by image cytometry. At least 150 tumour cell nuclei as well as 50 lymphocytes or trout erythrocytes for diploid reference were measured in each specimen. 35 morphological features, including those of the chromatin pattern, were derived from the digitized nuclear images. By means of multivariate discriminant analysis, significant differences could be found between tumours of 41 node-negative and 43 node-positive patients. The differences were sufficiently large to distinguish subgroups with low and high risk of lymph node involvement. Irrespective of lymph node status, 66 patients with an adequate 5 year follow-up were divided into two groups according to their survival time. The classification results revealed a clear relationship between characteristics of the nuclear image and prognosis. By image cytometry it is thus possible to separate both node positive and node-negative patients into subgroups with increasing risk of recurrence. The results indicate that nuclear image analysis provides prognostic information in addition to established prognostic factors, such as histological grade and tumour stage. PMID- 2560543 TI - Quantitative evaluation of morphological parameters for infiltrating ductal breast cancer prognosis. AB - The aim of our study was to determine the parameters intrinsic to ductal infiltrating carcinoma. 42 cases of about the same diameter (greater than 1.5 and less than 2.5 cm) with a minimum follow-up of more than seven years were selected. The nuclear and nucleolar shape parameters of one hundred cells for each case and the DNA content of 100 nuclei were measured by static cytometry. By means of multivariate discriminant analysis, the nucleolus/nucleus ratio and the DNA content proved to be the two best prognostic indicators based only on the examination of the parameters of the primary neoplasia. We were able to correctly classify 90% of the cases, and in particular to assign 100% of living cases and 81% of deceased ones to their exact categories. PMID- 2560544 TI - Minimum spanning tree, Voronoi's tesselation and Johnson-Mehl diagrams in human lung carcinoma. AB - A program written in Amba was developed for the automated construction of minimum spanning trees, Voronoi's tesselation and Johnson-Mehl diagrams, combined with quantitative measurements of nuclear parameters in histopathological specimens. Sections, 4 microns thick obtained from paraffin-embedded surgical lung specimens, were Feulgen-stained. The boundaries of the nuclei were calculated automatically after interactive identification of the relevant cells. The geometric centres of the nuclei were computed. They were used for constructing the minimum spanning tree, Voronoi's tesselation and the Diriclet cells. The direction of the main axis to the best fitting ellipsis according to the nuclear size was used for construction of the Johnson-Mehl diagrams. The following textural parameters were measured: 1) Minimum spanning tree: Distribution of neighbouring cells; minimum distance between neighbouring cells according to number of neighbours. 2) Voronoi's tesselation: Area and surface of obtained cells; Ratio of nuclear area/cell area. 3) Johnson-Mehl diagram: Area and surface of obtained cells; Ratio of nuclear area/cell area. The following nuclear parameters were measured: 1) Area, surface, absolute extinction (DNA-content); Similarity of quantitative nuclear parameters between neighbouring cells and in relation to number of neighbouring cells was analyzed. Twenty cases of primary lung carcinoma (five cases of each of epidermoid, adeno, large cell anaplastic, and small cell anaplastic carcinoma) were measured. Analysis of texture parameters in combination with morphometric nuclear parameters offers new possibilities in quantitative histopathology. PMID- 2560545 TI - Stereological estimates of nuclear volume in primary lung cancer. AB - The aim of this study is to provide unbiased, objective and reproducible morphometric data on primary lung carcinomas by using the stereological principle of estimating volume weighed mean volume of particles of arbitrary shape. The study includes 30 lung specimens; 10 from small cell carcinoma, 10 from squamous cell carcinoma, and 10 from adenocarcinoma. After standard fixation, embedding, sectioning and hematoxylin eosin staining, the mean nuclear volume was estimated using the formula: v upsilon = pi/3 . l0(3), where l0 is the length of the intercept through a test point hitting a nucleus measured in a random direction through the test point. The collection of the data took less than 10 min per specimen. The mean nuclear volume of small cell carcinomas varied from 109 to 238 microns 3 (group mean 152 microns 3), in squamous cell carcinomas from 407 to 826 microns 3, and in adenocarcinomas from 411 to 1046 microns 3 (group mean 736 microns 3). This simple and rapid method seems to provide unbiased, objective and reproducible data useful in the description and separation of primary lung carcinomas. PMID- 2560546 TI - Heterocycles as physiological ligands for the benzodiazepine receptor and for other binding sites. AB - Recently pharmacologically active benzodiazepines, including diazepam, have been identified in common foodstuffs, e.g. wheat and potato. The chronical intake by way of plant food may explain the existence of benzodiazepines in the brain and in other tissues of various mammalians and man. Hitherto these alkaloid-like compounds were considered to be merely products of industrial synthesis. All the benzodiazepines used in therapy show a similar chemical structure. However, depending on particular substituents, agonistic benzodiazepines can be subdivided into groups of different pharmacological potency. Besides benzodiazepines, in the past years other alkaloid drugs, e.g. derivatives of morphine, norharmane and tetrahydronorharmane, have been isolated from animals. Some of these substances have been discussed as physiological ligands of specific neuronal binding sites. Indications have been provided that at least part of these compounds or their precursors may be of plant origin too. The presence of these compounds in plants used for food may suggest complex interactions between plant and animal, exceeding the nutritional aspect. PMID- 2560547 TI - Pharmacological effects and binding studies of new methylxanthine thioderivatives. AB - The effects of two methylxanthine derivatives, 6-thiocaffeine (TC) and 6 thiotheophylline (TT), were investigated in different in vitro and in vivo conditions. On guinea-pig isolated trachea, both TC and TT showed a relaxant effect (EC50 50 microM and 60 microM, respectively), more potent than theophylline (300 microM). In guinea-pig isolated atria TC (30-50 microM) was able to antagonize R-PIA (a stable agonist on adenosine receptors) negative effect in not a clearly competitive way. Higher concentration (100 microM) began to reverse that inhibitory effect. In vitro Ki of TC and TT for A1 and A2 adenosine receptors was intermediate in comparison to caffeine and theophylline. On the contrary, the two thioderivatives showed a higher affinity for [3H] nitrendipine binding sites, in comparison to the original methylxanthines. All these data suggest a complex mechanism of action, probably at the level of adenosine extracellular receptors and L-type Ca2+ channels. In vivo experiments in mice provided evidence for a lack of CNS stimulant effects, but a loss of motor coordination was observed. Both thioderivatives showed a reduced acute toxicity. These data can be useful for the development of drugs for the therapy of asthma with reduced side effects. PMID- 2560548 TI - Effects of tetrahydrocannabinol content on marijuana smoking behavior, subjective reports, and performance. AB - This study investigated the smoking topography of marijuana and its effect on heart rate, subjective reports, and cognitive/psychomotor task performance. Male subjects (N = 12) with histories of moderate marijuana use smoked ad lib one cigarette containing 0, 1.3, or 2.7% delta 9-THC on separate days. Smoking topography measures revealed smaller puff and inhalation volumes and shorter puff duration for the high marijuana dose compared to the low dose. No other smoking behavior differed between the active doses. Heart rate was increased dose dependently over placebo levels. Active marijuana also increased subjective reports of drug effect over placebo, but not dose dependently. Significant memory impairment was observed on a forward and reverse digit span task, and performance was impaired on the digit symbol substitution task by the high, but not low, dose of marijuana. Performance on a divided attention task was not affected by marijuana. Thus, although subjects adjusted their smoking of cigarettes varying in THC content, dose-related effects of marijuana were obtained on several measures. The observed differences and individual variation in smoking topography measures suggest that precise control of smoking behavior would improve the accuracy of marijuana dose delivery. PMID- 2560549 TI - Inhibition of sleep and benzodiazepine receptor binding by a beta-carboline derivative. AB - The effects of systemic injections of beta-carboline-3-carboxylate-t-butyl ester (beta-CCtB) were investigated with regard to normally occurring sleep and several measures of benzodiazepine receptor occupancy in rats. A dose of 30 mg/kg of beta CCtB was found to have a long time-course of action as measured by an in vivo assay for benzodiazepine binding, with an 84% depletion of [3H]diazepam binding at one hour after the intraperitoneal injection. This dose of beta-CCtB was shown to delay sleep onset, decrease non-REM and total sleep in the first two hours after the injection, and to delay the appearance of REM sleep after the sleep onset. The dose- and time-dependence of the effects on sleep approximated the dose- and time-dependence of inhibitory effects of an IP injection of beta-CCtB on in vitro measures of benzodiazepine receptor affinity and number. PMID- 2560550 TI - [Non-invasive methods in the study of hemodynamics of Klippel-Trenaunay syndrome]. AB - In 10 patients with Klippel-Trenaunay syndrome, we have examined 13 extremities presenting characteristic lesions, using a mercury-gauged plethysmography. All cases had previously underwent arteriography and phlebography. The venous volume increased markedly in 12 of these 13 extremities, as well as the arterial flow in 6 extremities. The elevated skin blood flow was demonstrated in 4 of the 5 hemangiomas by Laser-Doppler velocity measurements. PMID- 2560551 TI - The role of free oxygen radicals in the mechanism of ischaemic-reperfusion myocardial injury. PMID- 2560552 TI - Effect of the beta-adrenergic substances propranolol, alprenolol and isoprenaline on cAMP production in bovine thyroid slices. AB - The effect of the beta-adrenergic blockers L-alprenolol and DL-propranolol and of the beta-adrenergic agonist L-isoprenaline on the basal and thyrotropic hormone(TSH)-stimulated cyclic adenosine-monophosphate (cAMP) level in bovine thyroid slices was studied. The main basal cAMP level in bovine thyroid slices was 3 pmol/mg tissue. TSH stimulated cAMP production in correlation to the concentration. Maximum stimulation was achieved with a TSH concentration of 10 mU/ml. The beta-blockers DL-propranolol and L-alprenolol caused 74 and 77% inhibition of TSH-stimulated cAMP synthesis respectively. The beta-adrenergic agonist L-isoprenaline did not significantly affect either the basal or the TSH stimulated cAMP level. The role of the beta-adrenergic receptor system in the regulation of TSH-stimulated cAMP synthesis is discussed. PMID- 2560553 TI - Phosphoinositide metabolism of rat pineal gland in a low ionic strength medium. AB - Pineal glands were incubated in the presence of 32P orthophosphate. When all NaCl in a conventional incubation medium was replaced by isotonic sucrose, i.e. when the ionic strength of the medium was decreased, there was a marked increase in 32P labelling of phosphatidylinositol (PI) and phosphatidic acid (PA). The 32P labelling of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) was not affected. No net synthesis of PI was observed. The increased labelling of PI therefore represents an increase in the turnover of PI. The 32P labelling of PI was observed also in media where NaCl was replaced by fructose or mannitol, but not in media, where NaCl was replaced by choline chloride. The effect depends on the concentration of the HEPES buffer and was not found in the medium with a bicarbonate buffer. 32P labelling of PI was not blocked by alpha 1 adrenergic blockers, phentolamine and prazosin, and did not depend on the presence of Ca2+ in the incubation medium. The effect was blocked by a Ca2+ channel blocker, MnCl2. Only 32P labelling of PI and not that of phosphatidylinositol 4-phosphate (PIP) and phosphatidylinositol 4,5-bisphosphate (PIP2) was increased during prolonged incubation in the sucrose medium. It is suggested that a decrease in the charge distribution across the plasma membrane as a result of the absence of most monovalent cations is responsible for the increased metabolism of phosphatidylinositol. PMID- 2560556 TI - Review of electron microscopic evidence favouring vesicle exocytosis as the structural basis for quantal release during synaptic transmission. PMID- 2560554 TI - Corticotropin-releasing factor (CRF) and interleukin-1 (IL-1) receptors in the brain-pituitary-immune axis. AB - Corticotropin-releasing factor (CRF) and interleukin-1 (IL-1) are two major components of the brain-endocrine-immune response to stress. We identified, characterized, and localized CRF and IL-1 receptors in brain, pituitary, and spleen using 125I-CRF and 125I-IL-1 alpha, respectively. 125I-CRF binding had comparable kinetic (KD:200-400 pM) and pharmacological characteristics in brain, pituitary, and spleen. In studies using cross-linking techniques, 125I-CRF was incorporated in rat pituitary and mouse spleen homogenates into a complex of Mr = 75,000 and in rat brain, into a complex of Mr = 58,000. The differences observed in the molecular weights between the pituitary and splenic vs. brain CRF receptors were evident across a variety of species and appeared to be due to differential glycosylation of the receptor proteins. In autoradiographic studies, CRF receptors were localized in highest densities in anterior pituitary and in brain regions involved in cognitive function, in limbic areas involved in emotion, and in brain areas regulating autonomic and other stress-related responses. In spleen, CRF binding sites were localized in the macrophage-rich red pulp and marginal zone surrounding the white pulp regions. High affinity 125I-IL 1 alpha binding sites were identified in pituitary membranes with kinetic and pharmacological characteristics comparable to the well-characterized IL-1 receptors in the EL-4 6.1 mouse thymoma cell line. Preliminary studies have identified IL-1 receptors in discrete regions of mouse brain. These data further substantiate the physiological role of CRF and IL-1 in modulating the brain endocrine-immune axis. PMID- 2560555 TI - Effects of glucocorticoid antagonism with RU 486 on pituitary-adrenal function in patients with major depression: time-dependent enhancement of plasma ACTH secretion. AB - Data from our group and others suggest that pituitary-adrenal activation in major depression reflects a defect at or above the hypothalamus which results in the hypersecretion of corticotropin-releasing hormone (CRH); some have suggested, however, that elevated indices of cortisol secretion and lack of suppressibility to dexamethasone may be a manifestation of a primary defect in glucocorticoid receptor activation. We report here a study of early morning pituitary-adrenal responses to the glucocorticoid antagonist RU 486 in patients with major depression and healthy volunteers. Previous data suggested that the response to RU 486 could represent an index of endogenous CRH secretory activity. RU 486 produced a robust increase in plasma corticotropin (ACTH) and cortisol secretion in both control subjects and depressed patients. In the controls, however, the increase was confined to the last 2 hours of sampling (6 to 8 am), whereas in the depressed patients the increase occurred throughout the sampling period (3 to 8 am). The ACTH response in the depressed patients exceeded that in the controls during most of the sampling period, including a significant (p less than .005) increase between 3 and 4:30 am. These results are compatible with the idea that hypercortisolism in major depression represents an alteration in the overall set point for hypothalamic CRH secretion rather than a primary alteration at the level of the glucocorticoid receptor. PMID- 2560557 TI - Patch clamp techniques used for studying synaptic transmission in slices of mammalian brain. AB - Procedures are described for recording postsynaptic currents from neurones in slices of rat brain using patch clamp techniques. The method involves cutting brain slices (120-300 microns thick) with a vibrating microtome followed by localization of cell somata, which can be clearly seen with Nomarski differential interference contrast optics in the light microscope. Tissue covering the identified cell is then removed mechanically and standard patch clamp techniques are applied. Using these methods, spontaneously occurring and stimulus-evoked inhibitory postsynaptic currents (IPSCs) were recorded from neurons in rat hippocampus at greatly improved resolution. In the presence of tetrodotoxin, to block presynaptic action potentials, spontaneous IPSCs seldom exceeded 25 pA. Evoked IPSCs elicited by constant electrical stimulation of a presynaptic neurone were larger and fluctuated in their amplitudes. Single-channel currents, activated by the putative inhibitory transmitter gamma-aminobutyric acid (GABA), had a size of about 1 pA. The number of postsynaptic channels activated by a packet of inhibitory transmitter is probably not more than thirty, nearly two orders of magnitude smaller than previously reported estimates for CNS synapses. This might reflect matching of synaptic efficacy to the high input resistance of hippocampal neurones and could be a requirement for fine tuning of inhibition. PMID- 2560559 TI - The use of porous hydroxylapatite granules in surgical treatment of advanced periodontitis. PMID- 2560558 TI - Receptor-controlled calcium discharge in frog heart cells. AB - A brief account is given of previous work concerned with the effects which ATP (P2-'purinergic') and alpha-adrenaline receptors have on the activity of the sarcoplasmic reticulum (SR) in frog heart atrial cells. More recent experiments made with single atrial trabeculae are also described which suggest, among other things, that ATP and alpha-adrenaline receptors both facilitate the process by which an action potential induces SR calcium discharge in atrial heart cells. The facilitatory effect involved is considerable; at high (greater than or equal to 2 microM) ATP doses, SR calcium discharge seems to be increased about 14-fold and, quite possibly, more than this. Results obtained with different levels of [Ca2+]o during ATP action suggest that the process of calcium-induced calcium release plays little part in the facilitatory process. Its putative mechanism is briefly discussed. PMID- 2560560 TI - [Experienced practice implant procedures. A standard determination]. PMID- 2560561 TI - [Physiology, precision technic and psychology. Position of dental prosthetics]. PMID- 2560562 TI - [AIDS--a practice risk for the dentist?]. PMID- 2560563 TI - [Surgical treatment of atrophic jaw for improved denture retention]. PMID- 2560564 TI - [Functional damage of occlusal system: examination findings and diagnosis]. PMID- 2560565 TI - [Clinical interpretation of relation between occlusal damage and muscle activity]. PMID- 2560566 TI - [Differences between intercuspidation in mouth, on articulator and on hand prepared model]. PMID- 2560567 TI - [Hydrolytic degradation of composites. 2. Sequence of use in practice]. PMID- 2560568 TI - [Factors favorable to accumulation and retention of plaque]. PMID- 2560570 TI - [Prevention and therapy of periodontopathies. New worldwide challenges to dentistry and health politics]. PMID- 2560569 TI - [Periodontal probe and its use]. PMID- 2560571 TI - [Antibiotic therapy as part of periodontal therapy]. PMID- 2560572 TI - [Sugar-free, tooth-friendly sweets from dental viewpoint]. PMID- 2560573 TI - [Free gingival transplants--an enrichment of restorative dentistry]. PMID- 2560574 TI - [Alkaline calcium hydroxide containing pulp capping material]. PMID- 2560575 TI - [Requirements in vitro for reconstruction of endodontically-treated molars]. PMID- 2560576 TI - [New adhesive restorations in functional therapy]. PMID- 2560577 TI - [Hemisection and crown placement in upper molars]. PMID- 2560578 TI - [Magnetic resonance in hepatic lesions. Experience with a 1.5-T magnetic field]. AB - Forty-one patients with histologically proven hepatic lesions (6 cysts, 6 hemangiomas, 8 hepatomas, 19 metastases and 2 negative cases) were studied with Magnetic Resonance (MR) imaging at 1.5 T, and with US and CT. This prospective study was aimed at evaluating: the comparative accuracy of MR, US and CT; the sensitivity and specificity of spin-echo (SE) vs FISP pulse sequences; the efficacy of T1 and T2 relaxation time values in differentiating hemangiomas from hepatomas and metastases. MR diagnostic accuracy was 94.7% vs 89.4% of CT and 84.2% of US. FISP sequences provided 60% sensitivity and 66% specificity. T2 relaxation time values were statistically significant (p less than 0.05) in differentiating hemangiomas (T2 range: 80.9-218.9 ms) from hepatomas (T2 range: 59.4-83.2 ms). The differences in mean T2 values between hemangiomas and metastases (T2 range 54.3-177.3 ms) were not statistically significant (p greater than 0.25). PMID- 2560579 TI - [Research on antibodies against BHV-1, BHV-2, BHV-4, BVD-MD virus, bovine adenovirus A and B, rotavirus and coronavirus in cattle in western Zaire: complementary results]. AB - Two-hundred bovine sera from western Zaire were screened for antibodies to 8 viruses: BHV-1, BHV-2, BHV-4, BVD-MD virus, bovine adenovirus A and B, bovine rotavirus and bovine coronavirus. Positive sera were found to all these viruses. For animals whose origin was undoubted, the main features were the high prevalence of infections by rotavirus and BHV-4 and the low prevalence of infections by coronavirus and BVD-MD virus. PMID- 2560581 TI - Prevention of pertussis. AB - Pertussis (whooping cough) is an acute respiratory disease caused by Bordetella pertussis. It occurs worldwide and is an important cause of morbidity and mortality in areas where immunization rates are low, particularly among children less than 1 year of age. The characteristic presentation of pertussis is paroxysmal coughing followed by a long inspiratory effort that produces the classic whoop. Lymphocytosis is frequently present. Complications include pneumonia and seizures secondary to hypoxia. The paroxysmal and convalescent stages of the illness can each last several weeks. Transmission occurs readily by respiratory droplets, and atypical or mild cases in older children and adults can be important in spread of the infection. Isolation, early erythromycin therapy, and erythromycin prophylaxis can reduce transmission, but vaccination is the primary means of control. An inactivated whole cell suspension of the bacterium has been an effective vaccine for protecting against pertussis since the 1950s, but whole cell vaccine may allow mild infections to occur and has been associated with local and systemic reactions that have eroded public acceptance. Component or acellular pertussis vaccines that are less reactogenic have been in use in Japan since 1981 and appear to be effective there. Development of an acellular preparation that is equally or more efficacious than whole cell vaccine may be possible, but clinical trials for measurement of protection against pertussis are difficult and trials with new pertussis vaccines will have to be carefully performed to avoid the controversies generated by earlier trials. PMID- 2560580 TI - rRNA gene restriction patterns of Legionella species: a molecular identification system. AB - A total of 28 species of Legionella could be differentiated by rRNA gene restriction patterns generated after cleavage of total DNA with either EcoRV or HindIII restriction endonucleases, and hybridization of fragments with 32P labelled Escherichia coli 16 + 23S rRNA. Different species gave different fragment patterns. When several isolates of a species were tested, the patterns obtained were often identical. However, more than one pattern was often observed when more than one serotype was considered. The method should be useful for the identification of all species of Legionella including those exhibiting immunological cross-reactions. PMID- 2560582 TI - Current use and future directions of adenovirus vaccine. AB - Adenoviruses are a major cause of respiratory illnesses in military recruits and also are common causes of respiratory and gastrointestinal infections during childhood. Forty-one serotypes of human respiratory and enteric adenoviruses have been identified. Live, oral adenovirus vaccines developed for the military and tested in large clinical trials have proved to be safe and highly effective in decreasing hospitalizations related to adenoviral acute respiratory diseases. Studies have demonstrated little horizontal transmission among military personnel but substantial transmission among family members. Use of recombinant techniques have opened new opportunities for the development of recombinant adenovirus vector vaccines against a number of viral pathogens such as hepatitis B, human immunodeficiency, herpes simplex and respiratory syncytial virus. PMID- 2560583 TI - [Role of radiotherapy in the treatment of small cell lung carcinoma]. AB - For several years, chemotherapy has been the only treatment of small-cell carcinoma of the lung, but radiotherapy is increasingly used for its local effect in various ways. Irradiation of the chest alternating or concomitant with chemotherapy has become a basic principle in localized forms of the disease and may perhaps be useful in its diffuse forms. The so-called "prophylactic" brain irradiation is routinely performed, even though its beneficial effect on survival has not yet been demonstrated. As for extensive irradiation, it is still under study. Radiotherapy therefore plays an important role which, added to that of chemotherapy, should result in an improvement of long-term survival in patients with small-cell carcinoma of the lung. PMID- 2560584 TI - [Pleuro-pulmonary localization secondary to adenoid cystic carcinoma of the sub maxillary glands]. PMID- 2560585 TI - Physiological role of cGMP and cGMP-dependent protein kinase in the cardiovascular system. PMID- 2560586 TI - Interaction of rheumatoid factor and Entamoeba histolytica. AB - The amoebae's cytotoxicity test and the amoebae's lysis test were used to show possible interactions between rheumatoid factor (RF) and Entamoeba histolytica. Amoebae's cytotoxic activity (ACA) was inhibited by affinity chromatography purified antiamoebae rabbit IgG (RIgG). Enhanced inhibition could be demonstrated with RIgG plus RF. But the same marked inhibition of ACA could be seen when replacing RF by heat inactivated normal human serum as a control. About 50% amoebae's lysis occurred when amoebae were brought together with native normal human serum (NNHS) as a source of complement. Amoebae's lysis increased to 60% when incubated with NHS plus human antiamoebae antibodies. No further augmentation could be obtained by the addition of RF. Using RIgG instead of human antibodies the lysis rate did not increase. Incubation of amoebae, NNHS, RIgG and RF even reduced amoebae's lysis. RF neither has an effect on ACA nor on complement mediated AL in vitro. PMID- 2560587 TI - A serological investigation of rotavirus infections in a shanty town population in Rio de Janeiro. AB - The presence of antibodies against rotavirus was investigated by enzyme immunosorbent assay (ELISA) in two distinct groups of children living in a shanty town in Rio de Janeiro. One hundred and thirty six plasma samples were randomly collected from children of 0 to 33 months (first group) and 255 serum samples were collected from other 85 children at ages of 2, 6 and 9 months (second group). A high percentage of antibodies were found in the newborn children and this rate decreased progressively until the age of 11 months, after which it increased again. At the age of 7 months, geometric mean antibody titers increased indicating that infection had occurred. PMID- 2560589 TI - [Status of the pituitary-adrenal cortex system in primary hypothyroidism]. PMID- 2560588 TI - [Adenosine cyclic 3',5'-monophosphate as a factor predicting the recurrence of cancer of the stomach]. AB - A method for the prediction of gastric carcinoma relapses has been developed and tried in 51 patients after a subtotal resection of the stomach. The method is based on the estimation of the ratio of cAMP levels in the remaining gastric mucosa along the resection line and in the tumor tissue. The accuracy of the method is 94.1%; its use permits a sufficiently reliable prediction of a gastric carcinoma recurrence. PMID- 2560590 TI - The effects of chronic treatment and withdrawal of CNS depressants on aggressive behavior. AB - Young adult CD-1 male mice were housed in individual cages throughout the study. Groups of 10 to 20 mice were given gradually increasing doses of delta-9 tetrahydrocannabinol (THC) at 6.25 to 25 mg/kg i.p., trifluoperazine (TFP) at 6 to 12 mg/kg p.o., phenobarbital sodium (PS) at 20 to 35 mg/kg p.o., morphine sulfate (MS) at 5 to 20 mg/kg i.p., methaqualone (MQ) 10 to 20 mg/kg p.o. or chlordiazepoxide (CDP) 10 to 25 mg/kg p.o. over four to six weeks to develop tolerance of these drugs. Following the development of tolerance, the drugs were withdrawn. On the fourth day of withdrawal, a young (3-4 weeks old) male mouse was introduced into the cage. When the intensity of the attack was measured by the percentage of animals that killed the intruder within four hours. The results indicated 0 to 4 percent in the controls, 50-54 percent for THC, 50 percent for TFP, 42 percent for PS and 57 percent for MS. In contrast, no killing behavior was exhibited by these mice after treatment with MQ or CDP. These data suggest that enhanced aggressive behavior, elicited by withdrawal from certain psychotropic drugs, may be measured by the killing (muricidal) behavior of isolated mice. PMID- 2560591 TI - [Ca2+-protease--an enzyme of the metabolism of cytoskeletal proteins in the olfactory membrane of vertebrates]. AB - Two forms of Ca(++)-activated protease (calpain I and calpain II) associated with an endogenous inhibitor (calpastatin) were detected in a cytosolic fraction of the olfactory tissue of vertebrates (pig, rat). Using ion exchange chromatography on DEAE-cellulose column, calpain I is divided into 2 peaks (eluting by 0.07-0.15 and 0.22-0.25 M NaCl), and calpain II is eluted by 0.35-0.40 M NaCl. The calpain activity was detected in fractions eluted by 0.1-0.17 M NaCl. The Ca(++) activated protease was demonstrated also in a fraction of cytoskeleton of olfactory tissue insoluble in a 1% solution of Triton X-100. The activity can be detected by Ca(++)-dependent destruction of exogenous substrate (casein), and by Ca(++)-dependent degradation of cytoskeletal endogenous proteins (16, 18 and 20 kDa), of which one may be calmodulin. PMID- 2560592 TI - [Ultrastructural study on the developmental process of the dentin bridge following direct capping using hydroxyapatite ceramic]. AB - Hydroxyapatite ceramics (HAP), which have been used in the endodontics or periodontics because of their bioafinitive characteristics, were applied directly as the capping of the wound surface after the experimental exposure of pulp in versional teeth to be extracted on orthodontics ground. In order to investigate the developmental process of dentine bridge induced by HAP capping, the pulp was studied ultrastructurally, 1, 3, 6 months after the treatment. One month after the capping, the multi-nuclear giant cells being rich in mitochondria and lysosomes were in contact with the HAP granules directly by extending their projections into the granules, which appeared to absorb the HAP granules. At the same time, the osteoblast-like cells with remarked rough endoplasmic reticulum (rER) were also observed just behind the giant cells. Only a few matrix vesicles and fibers were recognized around these osteoblastic cells. The HAP granules embeded in the pulp for 3 months were surrounded by fine matrix fibers which seemed to be produced by adjacent osteoblastic cells. The surrounding of these cells were filled with abundant matrix vesicles and collagen fibers, which would show the production of osteodentin matrix and the minerilization were active in this region. Six months after the treatment, the osteodentin matrix in upper dentin bridge became compact; matrix fiber surrounding the HAP granules became thick and a part of fibers inserted into the granules, which should mean that the HAP granules had bonded to the osteodentin matrix. In the deeper region of the dentin bridge, tubular dentin was formed newly. Both osteodentin and tubular dentin fused tightly without any organelles in the boader between them. The covering of hard tissue of dentin bridge then bonded to primary and reparative dentin closed the wound surface to protect the inner pulp. PMID- 2560593 TI - Associations of genetics and sampling time with levels of interleukin-2 activity. AB - The interleukin-2 (IL-2) assay has become a useful tool for examining the cellular immune response. Because of a lack of avian IL-2-dependent cell lines, the avian IL-2 assay in its present state, however, is not currently as powerful as its mammalian counterpart in effectively evaluating levels of IL-2 activity. The use of an avian IL-2 reference standard, Percoll-enriched populations of lymphoblasts, and sample collection times were examined to optimize the assay for comparing levels of IL-2 activity in a large number of birds. The reference standard was effective in accounting for assay-to-assay variance. Percoll enriched populations of responding lymphoblasts were more sensitive to IL-2 than nonseparated populations of cultured peripheral blood lymphocytes. Levels of IL-2 activity of cells isolated from the same bird did not change within a 1-week period, although differences did approach significance. In addition, levels of IL 2 activity differed between genetically distinct populations. PMID- 2560594 TI - Hepatoma presenting as extrahepatic biliary obstruction due to hemobilia. AB - A case of hepatoma presenting as extrahepatic biliary obstruction due to hemobilia is reported. The patient, a 49-year-old woman, developed jaundice of the obstructive type after a history of B-viral hepatitis. On laparotomy, the liver revealed macronodular cirrhosis without any noticeable mass. A 4-cm sized friable tissue and blood clots were identified within the distended left hepatic duct. Pathologic examination of this tissue confirmed the diagnosis of hepatocellular carcinoma extended in the hepatic duct. PMID- 2560596 TI - Characteristics of a prompt morphodynamic response of the pineal gland to an acute ACTH injection. AB - The morphodynamic response of the pineal gland in the period 5-30 min after a single ACTH injection was studied. In 5 min a saline injection, functioning as a control, elicited a prompt release of the pineal active compounds advocated to be contained in clear vesicles and structures traditionally labeled lipid droplets. In the subsequent time period, 15 min, a rise in the relative volume of the granular endoplasmic reticulum (GER) and the Golgi apparatus, as well as prosecretory granules, was interpreted as a sign of an increased pinealocyte activity which leads to the restoration of the pineal gland endocrine potential. A decline in the increased mass of this membranous elaborative compartment in 30 min could be understood as an illustration of the stabilisation of the pinealocyte stress-evoked response. The ACTH injection did not apparently change the pattern of the morphodynamic pinealocyte response promoted by the saline injection in 5 min. However, starting from 15 min, a significant increase of the mass of GER and the Golgi apparatus was found in both pinealocyte populations. The augmented presence of prosecretory granules and lipid droplets, in comparison with the saline-injected animals in the respective time period, as well as the absence of a clearly observable evidence of the pinealocyte secretory activity pointed out that the ACTH stimulative action on the pinealocyte membranous elaborative compartment rather facilitated the pinealocyte endocrine recovery than supported their current secretory activity. From the functional viewpoint, the results obtained suggest that a stress-induced ACTH burst could modulate the post-secretory restitution of the endocrine potential of previously stress activated pinealocytes. PMID- 2560595 TI - [Gastrointestinal cytomegalovirus manifestations in AIDS]. AB - Cytomegalovirus (CMV) infection is one of the most important intestinal opportunistic infections in AIDS. In severe cases ulcerations and colitis are the commonest manifestations. 184 HIV positive patients with gastrointestinal symptoms were investigated by endoscopy of the gastrointestinal tract. While culture, immunohistochemical staining and histology from biopsies were performed, the results of all three methods were compared. In one third the cases CMV associated lesions could be found by endoscopy. Erosions or ulcerations are the most frequent tissue lesions. In 95% the culture was positive. In addition, immunohistochemical staining in 75% and histology in 61.7% were positive in patients with more serious manifestations. For early diagnosis endoscopy of the gastrointestinal tract and histological, histochemical and microbiological investigations of biopsies are essential. PMID- 2560597 TI - Immuno- and histochemical evidence for changed neurotransmission in basal forebrain, hippocampal formation and neocortex of aged rats. AB - The distribution of choline acetyltransferase- and gamma-aminobutyric acid immunoreactivity as well as acetylcholinesterase-, gamma-aminobutyrci acid transaminase- and monoaminooxidase-positive structures in the basal forebrain, hippocampal formation and neocortex of young adult and aged rats was compared. The changes in the staining and density of the immuno- and histochemical positive nerve cells and fibers observed provide evidence for the involvement of different transmitters in normal ageing. PMID- 2560598 TI - Acetylator phenotype and short-course chemotherapy for tuberculosis in Papua New Guinea. AB - In Papua New Guinea the slow acetylator phenotype is unusual and peripheral neuropathy during isoniazid therapy is uncommon. Acetylator status and other factors influencing isoniazid-induced peripheral neuropathy are discussed, and statistics from Papua New Guinea are presented. The relevance of acetylator phenotype to other side-effects of isoniazid is less well defined. Prophylactic pyridoxine supplementation during short-course chemotherapy for tuberculosis is not generally necessary in the daily 'A' regimen, but should be maintained in the twice-weekly (high-dose isoniazid) 'B' regimen for adults and children over 20 kg. PMID- 2560599 TI - [Alveolar bone reconstruction with hydroxyapatite in aged humans]. AB - There is a rising need for preprosthetic surgery due to a general increase in life expectancy and the desire for optimal oral rehabilitation. A method of reconstructing residual ridges in an ideal form with a hydroxylapatite-fibrin paste is presented. Patients suffering from mild to moderately severe age-related neurological disorders with dementia and psychomotor slowing benefit from the procedure, whereas psychiatric illnesses such as hypochondria or psychotic disturbances need to be excluded from ridge reconstruction. The same is true for neurological deficits resulting in facial and oral hyperkinesia. PMID- 2560600 TI - [Investigation of utility and mechanics of use of hydroxyapatite for therapy of hypersensitive tooth root]. AB - The efficiency of extremely small particles of hydroxylapatite on a therapy with hypersensitive tooth-necks was approved in a double-blind-study. 90% of the test persons stated an improvement of their complaints after 3-5 days, and 50% became complaintless within the time of study. The manner of working was proved with the help of x-ray inclination and by polarisation-optical studies. PMID- 2560602 TI - [Comparison of two powder-stream systems for tooth polishing]. AB - The effect of two airpolishing systems on the surface roughness of polished ceramic test pieces was compared and evaluated using standardized procedures. Test pieces resembling enamel in hardness were treated with these air-polishing systems. Using a Perthometer surface roughness was found to increase significantly. Thus the possible effects of airpolishing systems on the enamel surface should be considered. Efficiency of extrinsic stain removal was studied by SEM and documented. Photographic techniques provided information on the homogeneity of the sodium bicarbonate spray. PMID- 2560601 TI - [Hydroxyapatite use in dental practice. 2. Repair of bone defects in dental and jaw surgery operations]. PMID- 2560603 TI - [Distribution and lateral mobility of concanavalin A receptors in the course of cell cycle of human gastric cancer cells]. AB - The distribution and lateral mobility of Con A receptors in the course of the cell cycle has been studied on human gastric low-differentiated mucous adenocarcinoma cells (MGC 80-3). Synchronized MGC 80-3 cells in monolayer were labeled with F-ConA in the different phases of cell cycle. The discontinuous pattern of surface labelling was present during G1, S and G2, while there were marked differences among them. The cells in mitosis displayed uniform distribution of Con A receptors complex with strong fluorescence. The fluorescence photobleaching recovery method has been used to determine the lateral mobilities of Con A receptors complex. There was marked difference among various phases for mobility pattern and diffusion coefficient. The pattern of mobility of Con A receptor complex was appeared diffusion mobility in M and G1 phases, while flow pattern occurred in S and G2 phases. The diffusion coefficient (D) G1 greater than M, the rate of flow S greater than G2, but mobile fraction in G2 phase is higher than other phases. These results showed that dynamic properties of Con A receptor are apparently different which were regulated by the cell cycle. PMID- 2560604 TI - [EGFR expression and EGF stimulation of proliferation in human liver carcinoma cells]. AB - Epidermal growth factor receptor (EGFR) gene expression and growth stimulation of EGF on human hepatoma cells of cell lines BEL-7404 and SMMC-7721 were studied. 125I-EGF binding assay was used to measure the binding characteristics and the amounts of EGFR on these cells. The binding time course and the binding competition assay showed that the binding of 125I-EGF to 7404 cells was saturable and specific. Scatchard analysis of EGF binding curve indicated that 7404 and 7721 cells expressed approximately 1.1 x 10(5) and 0.7 x 10(5) EGFRs per cell with binding affinity (Kd) 2.1 nM and 1.8 nM respectively. Northern hybridization and immunoblotting analysis showed the EGFR gene expression products in 7404 and 7721 cells were 5.6 Kb mRNA and 170 Kilo-dalton glycoprotein. Anchorage-dependent growth of 7404 and 7721 cells was stimulated in the presence of nanogram quantities of EGF in medium containing 10% calf serum or 0.5% calf serum. The factors in serum appeared to act synergitically in stimulating of cell proliferation. EGF also stimulated the anchorage-independent growth of 7404 and 7721 cells in soft agar. The results suggest that EGFR is actively expressed in human hepatoma 7404 and 7721 cells and EGF may be one of the mitogens needed for the growth of hepatoma cells. PMID- 2560605 TI - Diet and chronic diseases: a global perspective. PMID- 2560606 TI - Defective immune response to Epstein-Barr virus in patients with acute lymphocytic leukemia. AB - Thirteen children with acute lymphocytic leukemia (ALL) receiving maintenance chemotherapy were tested for anti-Epstein-Barr virus (EBV) antibodies, EBV specific cytotoxic T lymphocyte (EBV-CTL) activity and spontaneous cytotoxicity against Raji cells in order to define the defective cellular immunity in ALL children during complete remission. Among 10 seropositive patients, anti-virus capsid antigen (VCA) antibody titer varied from 1:20 to 1:320 and anti-EBV nuclear antigen (EBNA) antibody was not detectable in four. No patients were positive for anti-early antigen (EA) antibody. EBV-CTL activity and interleukin 2 (IL-2) or interferon alpha (IFN-alpha) augmented EBV-CTL activity in eight seropositive patients was significantly lower than that in the seropositive age matched control group. IFN-alpha, OK-432 or IL-2 augmented spontaneous cytotoxicity were also significantly lower in the patients compared to those in the control group. These defective killer cell activities may allow EBV infections to enter a severe, fulminant or persistently active state in the patients with ALL receiving aggressive maintenance chemotherapy. PMID- 2560607 TI - Chemoattractant receptors of neonatal polymorphonuclear leukocytes: lack of modulation of the receptors by membrane modifier. AB - To analyze a defect(s) in chemotactic responsiveness of human neonatal polymorphonuclear leukocytes (PMN), PMN functions and their chemoattractant receptor properties were examined using N-formylmethionyl-leucyl-phenylalanine (FMLP). Amphotericin B (AmB) is known to affect human PMN membrane and to alter cellular functions. Although the AmB treatment decreased the FMLP binding and cellular movement in adult PMN, there were no such changes in neonatal PMN. Kinetics studies of 3H-FMLP binding to PMN, furthermore, indicated the poor internal transportation of the chemoattractant into neonatal PMN. These abnormalities, which are related to the chemoattractant receptors, may explain in part the defective chemotaxis of neonatal PMN. PMID- 2560608 TI - Effects of inflammation on copper antioxidant enzyme levels. AB - Inflammation increases plasma levels of ceruloplasmin, a copper protein with possible antioxidant function. This paper describes modulation of these increases by copper intake, and describes combined effects of inflammation and copper intake on Cu-Zn and extracellular (EC) superoxide dismutase (SOD) activities. Turpentine injections in rats fed 1 of 4 copper levels increased ceruloplasmin activities, but values were sensitively limited by copper intake. Cu-Zn SOD activities in the liver, but not in erythrocytes or lungs, were reduced by inflammation in each dietary copper group. Inflammation in rats fed a standard mixed feed diet reduced plasma EC superoxide dismutase activities measured by inhibition of pyrogallol autoxidation. Different results were obtained with 3 xanthine oxidase based SOD assays which were each subject to assay interference. Studies in humans found a group of rheumatoid arthritis patients to possess relatively low erythrocyte SOD and relatively high ceruloplasmin activities. Activity levels of SOD, but not of ceruloplasmin, increased after 4 weeks of copper supplementation (2 mg/day). The fate of cellular Cu-Zn SOD activity contents in inflamed tissues is largely uninvestigated. However, interleukin-1, a hormone released at inflammation sites, elevated Cu-Zn SOD activities in cultured fibroblasts. PMID- 2560609 TI - Inhibitors of copper bioutilization: fiber, lead, phytate and tannins. AB - Considerable evidence suggests that copper intakes of Americans have declined in recent years to levels that might be considered seriously low. Under such conditions, other dietary factors which inhibit copper utilization might be of concern since otherwise marginally adequate copper diets might be made copper deficient. A large number of mineral nutrients have been found to interact with copper. Non-nutrient substances in food also may affect copper utilization. Evidence suggesting that lead, certain kinds of dietary fiber, phytates and tannins inhibit copper utilization are reviewed in this paper. PMID- 2560610 TI - Chemical composition and vitamin B content of abattoir by-product meals. AB - Chemical composition of meat-and-bone meals (n = 22), poultry by-product meals (n = 12), blood meals (n = 6), bone meals (n = 4) and feather meals (n = 8) indicated a slight degree of overdrying. Crude fat and ash contents were within the acceptable limits. Mineral elements of bones were determinative in the element pattern of meat-and-bone meals as indicated by the significant correlations between Ca, P and Mg. Crude protein was in negative regression and correlation with crude ash content (r = -0.81; P less than 0.001). This was more pronounced when the data were expressed in fatless dry matter (r = -0.94; P less than 0.001). Crude protein also showed significant negative correlations with Ca, P and Mg contents, offering the possibility to predict their levels from nitrogen content. Meat-and-bone meals and poultry by-product meals were relatively rich sources of riboflavin, niacin, pantothenic acid and biotin, whereas their pyridoxine and thiamine contents were low. Blood meals, bone meals and feather meals were found to be negligible vitamin B sources. Vitamin B and crude protein content of abattoir by-product meals showed uncertain correlations. PMID- 2560611 TI - [Contraction of skeletal muscles: regulation of calcium intracellular movements]. AB - The different membrane systems and proteins involved in the control of intracellular calcium movements in the skeletal muscle cell are described. These include the sarcoplasmic reticulum, that Ca(++)-ATPase sarcoplasmic reticular calcium pump, transverse tubules, calcium channels, and the ryanodine receptor protein. The significance of these systems is shown clearly in the myopathies, where the main errors involved do not concern the contractile system, but the command and control mechanisms. PMID- 2560612 TI - [Screening tests for malignant hyperthermia susceptibility]. AB - The ideal screening test for malignant hyperthermia susceptibility (MHS) has yet to be discovered. It should be simple noninvasive, yet totally specific and sensitive. Until such an ideal test becomes available, allowing simple routine preoperative screening, tests should only be used in certain specific situations. These include: patients in whom a clinical crisis was suspected; the members of the family of a subject labeled MHS because of a fatal, or otherwise, crisis, or in whom tests were positive; patients with other pathological conditions which could be linked to malignant hyperthermia (MH) (some myopathies, effort or stress MH, neuroleptic malignant syndrome). The various tests proposed in the literature aim at revealing MHN subjects, using or not a triggering agent, halothane most often. However, detecting these abnormalities sometimes gives greater insight into the physiopathology of MH than in the detection of an individual patient's susceptibility. The tests have been classified as in vivo, electrophysiological, blood, and in vitro muscle biochemical, morphological, and pharmacological tests. The discovery of new tests gives renewed hope: CPK levels, platelet tests, calcium sarcoplasmic reticular reuptake, lymphocyte Quin 2 test, nuclear magnetic resonance spectroscopy. However, experts worldwide agree that the only reference test to this day remains the in vitro halothane caffeine contracture tests. These tests have shown their reliability; they must be performed on muscle strips obtained from surgically removed muscle biopsies, by laboratories used to this technique and who have at their disposal a sufficiently large group of MHS subjects with a clear-cut clinical crisis, as well as controls. The patients must therefore travel to these laboratories. The design of common protocols for European laboratories on one hand, and the North American laboratories on the other, is a good guarantee of the reliability of these tests. PMID- 2560613 TI - [Parathyroid insufficiency syndrome. Two new cases: type Ia pseudohypoparathyroidism and pseudo-idiopathic hypoparathyroidism]. AB - Two new cases with parathyroid insufficiency syndrome are described. The first one is a seven year and 6 months old male with Pseudohypoparathyroidism Type Ia, who has presented: hypocalcemia, hyperphoforemia, increased PTH and TSH, prolactin decreased and Albright hereditary osteodystrophy phenotype. The second one in a six year and four months old female with Pseudo-idiopathic hypoparathyroidism who has presented: hypocalcemia, hyperphosforemia, increased PTH, without any hormonal disturbances nor dysmorphias. Ellsworth-Howard test with 50 micrograms of 1-34 hPTH is made in both, remarking the different renal response about phosphaturia and plasmatic, urinary and nephrogenic cyclic AMP. The treatment is performed in both with calcitriol and oral calcium (L-Thyroxine is associated for the first patient). Biochemical disturbances are normalized. Other parathyroid insufficiency syndrome are revised. PMID- 2560614 TI - [Hepatoblastoma: echotomographic evaluation. Apropos of a case]. PMID- 2560615 TI - Worksite inspection and the control of occupational disease. The OSHA experience. PMID- 2560616 TI - Retinopathy before the diagnosis of AIDS. AB - We reviewed the records of ocular examinations of patients referred or examined for inflammatory retinal disease over a two-year period and found 18 cases in which retinopathy was documented before the diagnosis of acquired immunodeficiency syndrome (AIDS). Seventeen patients were either homosexual men or intravenous drug abusers. Although 13 patients had AIDS-related complex, no patients met the Centers for Disease Control criteria for AIDS before their ophthalmologic examination. In all 18 cases, the diagnosis of severe immunodeficiency suggestive of infection by the human immunodeficiency virus (HIV) was made by the ophthalmologist. These ophthalmologic findings included five patients with asymptomatic retinal cotton-wool patches, two patients with endogenous Staphylococcus epidermidis bacterial endophthalmitis, and 11 cases of isolated cytomegalovirus retinitis. All patients were ambulatory outpatients at the time of ophthalmologic examination. Noninfectious retinopathy and intraocular opportunistic infections suggest the diagnosis of HIV infection and AIDS, and the ophthalmologist may play an important role in early diagnosis of this disease. PMID- 2560617 TI - Persistent active herpes virus infection associated with atypical polyclonal lymphoproliferation (APL) and malignant lymphoma. AB - This study focuses on lymphoproliferative diseases associated with persistent infection by Epstein-Barr virus (EBV) and human herpes virus 6 (HHV-6). A suggestive premalignant lymphoproliferative state is distinguished from malignant lymphoma and identified as "atypical polyclonal lymphoproliferation" (APL). Sixteen cases of herpes virus (HHV)-associated APL are compared with 21 cases of HHV-associated malignant lymphomas (ML), with 108 cases of EBV or HHV-6 related acute infections mononucleosis, with 14 cases of seronegative non-specific lymphoid hyperplasia and with 304 cases of HHV-unrelated ML. Six cases of APL and two ML occurred in AIDS patients, two cases in Sjogren's syndrome, one in a kidney allograft recipient, and the remaining cases had no identified underlying disease. APL was histologically reminiscient of excessive infectious mononucleosis, while other cases of Castleman's disease or even of malignant lymphoma. Seropositive APL and ML contained significantly more virus genome than is found in latent background infection. There was no histologic difference between HHV-6 or EBV-positive APL or ML, although both viruses infect different lymphocyte populations. From histology alone, seropositive ML cases were not distinguished from seronegative ones, yet persistent active EBV and HHV-6 appear to predominate in follicular center cell- and immunoblastic lymphoma and in HOdgkin's disease. Although no a direct oncogenic activity of these viruses could be observed in our cases, they may contribute to lymphomagenesis by inducing progressive lymphoproliferation. PMID- 2560618 TI - The effect of iron on experimental colorectal carcinogenesis. AB - The effect of parenteral and oral iron was examined in the rat 1,2 dimethylhydrazine (DMH) colorectal carcinogenesis model in a series of experiments. Parenteral supplementation of iron was found to augment tumor yield (p = 0.012) and oral iron was found to augment tumor incidence (p = 0.03, when control groups were combined). In addition, phytic acid, a significant component of dietary fiber was found to reverse the augmenting effect of oral iron on tumor yield and incidence (p = 0.09 for both). Furthermore, in a short term DMH nuclear toxicity assay, analysis of the karyorrhectic index (KI), there was no difference in the KI between oral iron and phytate dietary groups (p = 0.53 for the left colon and p = 0.2 for the right colon), implying that iron's effect on colorectal tumor induction takes place during the promotional phase of carcinogenesis and not during initiation. These experiments support the epidemiologic observation that dietary iron may augment colorectal cancer risk and that the mechanism by which dietary fiber diminishes colorectal cancer risk may be the chelation of dietary iron by the phytic acid component of dietary fiber. PMID- 2560619 TI - Monoclonal B cell proliferation in lymphoproliferative disease associated with herpes virus type 6 infection. AB - Clonal gene rearrangements and aneuploidy are commonly recognized as characteristics of malignancy. We challenge this by an observation of a 3 1/2 year old boy with chronic benign lymphadenopathy, connatal splenomegaly and peripheral lymphocytosis. His brother and mother suffered from similar disease without progression. Cervical lymph node biopsy from our patient revealed a benign histology corresponding to the clinical course; however, clonal rearrangements of the JH and TCR gamma gene were demonstrated in lymph node cells and DNA aneuploidy in 11% of bone marrow cells. Immunohistology showed an inverse T-helper/T-suppressor cell ratio with polyclonal B cell increase and extensive lymphocyte activation. In situ hybridization demonstrated foci of lymphoid cells in the paracortical zone with expression of herpes virus type 6 genome. In addition, the patient, his brother and his mother showed serological evidence of active infection with the human herpes virus-6 (HHV-6). The association of HHV-6 infection, monoclonality and benign lymphoproliferation is discussed. PMID- 2560620 TI - Molecular species of the antitumor and antiviral fraction from pine cone extract. AB - The most active fraction (Fr. VI) of antitumor and antiviral pine cone extract was subjected to chemical structure analysis. IR, UV, and ESR spectroscopies, in addition to chromatographic analysis on cellulose-TLC plates, reveal that the major component of Fr. VI includes structures belonging to the lignin family. A commercial alkali-lignin showed antiviral activity and granulocyte iodination stimulating activity as effective as those of Fr. VI, but had much weaker immunopotentiating activities, such as antitumor and antimicrobial activities and splenocyte stimulating activity. As a tentative conclusion, the active principle of Fr. VI might be complex(s) of lignin-like polyphenolic skeleton with a molecular matrix of unknown components. PMID- 2560621 TI - Optimal treatment for T1-3NOMO small cell lung cancer: surgery plus adjuvant chemotherapy. AB - To improve treatment results in patients with surgically resectable T1-3NOMO small cell lung cancer, 16 patients were randomized to receive either neoadjuvant (preoperative) or adjuvant (postoperative) chemotherapy (cyclophosphamide, epirubicin, and etoposide). The overall survival is 45% at a median follow-up of 28.5 months. Patients treated with surgery plus adjuvant chemotherapy (n = 8) showed a significantly higher median and overall survival, proportion of 2-year survivors, and lower systemic relapse rate. These results indicate that the optimal treatment for T1-3NOMO small cell lung cancer is represented by the combination of surgery plus adjuvant chemotherapy. PMID- 2560622 TI - Tumorigenicity assay of the human mammary carcinoma cell line MCF-7. AB - A tumorigenicity assay was performed using DNA extracted from the human mammary carcinoma cell line MCF-7. Seven nude mouse tumors were obtained and were analyzed for the presence of human sequences and known activated oncogenes. The c erbB.2/neu gene was identified in two tumors and two ras oncogenes in two other tumors. In order to characterize the mechanism of activation of the c-erbB.2/neu oncogene, we isolated and analyzed this gene from one of the tumors. The cloned gene did not present any rearrangement. c-erbB.2/neu transcripts from the nude mice tumors were of normal size. In one case overexpression was observed. PMID- 2560624 TI - Neuron-specific enolase in small-cell carcinoma of the lung: the value of combined immunocytochemistry and serum determination. AB - The value of neuron-specific enolase (NSE) as a marker for small-cell carcinoma of the lung has been the subject of several reports. Taken together, about 70 per cent of patients with small-cell carcinoma had a raised serum concentration of NSE, and in a majority of patients NSE could be detected immunocytochemically in tumor biopsy speciments. This study examined the diagnostic value of combined immunocytochemical detection and serum determination of NSE in 96 unselected patients with small-cell carcinoma. Seventy-one patients had raised serum concentrations and in 69 a positive immunoreaction for NSE was demonstrated in the biopsy cells. However, for 87 (91%) of the patients, both or either of the NSE assays were positive. The combined use of immunocytochemistry and serum determinations thus gave better information on tumor NSE expression than either method alone. When patients with small or mechanically maltreated biopsy specimens were excluded, the accuracy of the combined assays was even higher (47/49, 96%). We therefore conclude that NSE, although not a specific marker for small-cell carcinoma of the lung, is useful as a complement to conventional diagnostic procedures and, when assayed both in biopsy material and in patient sera, tumor NSE expression can be demonstrated with a high precision. PMID- 2560623 TI - Studies on factors influencing human plasma alpha-MSH. AB - The various physiological effects of alpha-MSH, mainly on the CNS and on pigmentation in animal models, are well documented in the literature. Only a few investigators have confirmed similar properties in the human. However, the possible physiopathological role played by this hormone in human melanoma is still poorly defined. In order to approach this subject in a manner as complete as possible, we have performed, during the past four years, three different series of experiments: 1) alpha-MSH measurements in plasma samples from: a. melanoma and other cancer patients, b. whole body UVA irradiated healthy adults, c. circadian rhythm determinations in melanoma patients and in healthy male adults; 2) alpha-MSH measurements in human melanoma tumours; 3) alpha-MSH receptor expression on human melanoma cells in culture involving: a. alpha-MSH radio-binding assays and b. tyrosinase assay. Our results so far show 1) increased alpha-MSH levels in melanoma patients' plasma, alpha-MSH responsiveness to UVA stimulated skin, large immunoreactive alpha-MSH content in melanoma metastases and an alpha-MSH circadian rhythm in some individuals different from cortisol; 2) alpha-MSH receptor expression in melanoma cells could be increased by various effectors able to stimulate melanogenesis. PMID- 2560625 TI - Effects of interferon-beta on Daudi cells and on small cell lung carcinoma cells which over-express the c-myc oncogene. AB - The effects of human interferon-beta (IFN-beta) on four lung cancer cell lines, two of which over-express the c-myc oncogene, were compared to Daudi cells which also over-express c-myc. The anti-proliferative effect of IFN-beta on Daudi cells was correlated with a decrease in steady state levels of c-myc mRNA. IFN-beta treatment of a classic small cell lung carcinoma cell line and a non-small cell lung carcinoma cell line which express normal levels of c-myc did not decrease the rate of cell proliferation. IFN-beta, however, was found to have dramatic anti-proliferative effects on the two lung cancer cell lines with c-myc over expression. The growth inhibitory effects of IFN-beta on the two lung cancer cell lines could not, however, be correlated with decreased amounts of c-myc mRNA. Furthermore, Western blot analysis showed that interferon treatment did not detectably affect the c-myc protein levels in these cells. Therefore, the mechanism by which IFN-beta slowed the growth rates of the two lung cancer cell lines did not appear to directly involve down-regulation of the c-myc oncogene, expressed in amplified amounts, in these cells. PMID- 2560626 TI - Specific receptors for 1,25-dihydroxyvitamin D3 (1,25-DR) and human colorectal carcinogenesis. AB - Because it is a common prerequisite for steroid responsiveness in target tissue, we investigated the presence of specific 1,25-DR in spontaneous human colorectal adenocarcinomas (ADC) and adjacent normal-appearing mucosa (NAM) from 23 operative specimens (12 male and 11 female patients). 1,25-DR was determined in cytosol by a DCC assay technique. 1,25-DR was present in 21 of 23 NAM and in only 4 of 23 HCRA. All positive ADC were well differentiated. Receptor content expressed in femtomoles/mg of protein (mean +/- SEM) was respectively 63.9 +/- 7.6 for right colon NAM and 51.3 +/- 12.9 for left colon or rectum NAM. When we compared all NAM specimens, receptor content was 56.7 +/- 8.0 femtomoles/mg of protein. No difference in 1,25-DR NAM level was observed between right colon and left colon or rectum. In adenocarcinoma the mean content was 66.5 +/- 14 fmoles/mg of protein. Scatchard analysis showed a single class of specific high affinity saturable 1,25-DR with a dissociation constant (Kd) of 0.97 +/- 0.57 and 1.03 +/- 0.39 chi 10(-10) M in NAM and ADC respectively. These preliminary data represent the first demonstration of 1,25-DR throughout the entire human colon and indicate that the receptivity for this hormone is often lost during malignant transformation of the human colorectal mucosa. In addition, 1,25-DR could be a marker of differentiation in ADC. These preliminary results provide evidence supporting the addition of Vitamin D to the roster of developmental cancer chemopreventative agents. PMID- 2560627 TI - Relationship of the 1 alpha, 25-dihydroxyvitamin D3 receptor concentration and the DNA content in renal cell carcinomas. AB - A role has been suggested for 1 alpha,25-dihydroxyvitamin D3 (1,25-(OH)2D3) in cell proliferation and differentiation. Therefore the concentration of the 1,25 (OH)2D3 receptors and DNA-ploidy was measured in 22 renal cell carcinomas. No relation was found, the mean 1,25-(OH)2D3 receptor concentration being 8.4 fmol/mg of protein (range 2.8-15.9) in diploid tumors and 7.0 fmol/mg of protein (range 0-27.8) in DNA aneuploid tumors. The aneuploid tumors (11 out of 22) had a heterogeneous DNA content in 9 out of 11 cases. At the time of operation, no patient had metastases. In this prospective study, one out of 9 patients with DNA aneuploid tumor had died of renal cell carcinoma (observed 33-58 months, median 42 months). None of 10 patients with diploid tumors had died (observed 40-56 months, median 52 months). PMID- 2560628 TI - [The energy utilization of coarse feed by adult swine]. AB - The energetic utilization of 12 technically dried roughages (6 legumes, 5 grasses, 1 straw) with a wide variation of the chemical composition (crude fibre content 190-395, crude protein content 61-266 g/kg DM) was tested with 8 adult pigs each by means of the respiration test technique. The experiments were laid out according to the difference method with 3-4 basal ration period per animal. The digestibility of the energy of the feedstuffs (except straw) ranged from 40 60%, the partial utilization of the metabolizable energy from 63 to 74%. PMID- 2560629 TI - Nucleotide ectoenzyme activities of human and Chinese hamster fibroblasts in tissue culture. AB - We have previously assigned human ecto-5'-nucleotidase (NT) to chromosome 6 on the basis of conversion of exogenously supplied [14C]AMP to adenosine by whole cells of human and Chinese hamster hybrids carrying chromosome 6. In this paper we demonstrate that the activity on human MRC-5 fibroblasts is typical of previously described and purified ecto-5'-nucleotidases. In contrast to MRC-5 cells, Chinese hamster V79A2 cells weakly express an AMPase activity that is not NT. The cytosolic form of NT in human and hybrid fibroblasts is similar to the ectoenzyme in substrate specificity. Hybrids that lack chromosome 6 express neither the ecto- nor the cytosolic enzyme, suggesting that both forms may be coded by the same gene on chromosome 6. Ecto-ATPase, ecto-ADPase, and ecto-ADP kinase activities are each expressed at similar levels in MRC-5 and V79A2. The ATPase, ADPase and NT activities of MRC-5 cells act sequentially to generate adenosine. A similar cascade acts on V79A2 cells but the lack of NT causes the accumulation of AMP. PMID- 2560631 TI - Genetic variation in adenylate kinase 1, glyceraldehyde-3-phosphate dehydrogenase, and glutamic-pyruvate transaminase in the marsupial Monodelphis domestica. PMID- 2560632 TI - Alcohol and thiamine of the brain. PMID- 2560630 TI - Four tandem defective P elements associated with positive regulation of the Drosophila melanogaster glucose-6-phosphate dehydrogenase gene. AB - Three high-glucose-6-phosphate dehydrogenase (G6PD)-activity mutants (2512H, S44H, and 1FH) are characterized by two insertion sequences associated with the G6PD locus; one (Ins1; 3.5 kb long in 2512H and S44H and 2.9 kb long in 1FH) is present just 5' to exon I and consists of a KP' (the 32nd base of the KP was replaced by guanine), a core sequence and a KP, and the other is 4.2 kb long and resides within an intron. Southern blot analyses of revertants showing low G6PD activity suggested that the insertion sequence responsible for high G6PD activity may be the core sequence but not the flanking KP and KP' or the Ins2. DNA sequencing data of the clone carrying the core sequence of 2512H demonstrated that the core sequence is another type of defective P elements (core P). Interestingly, a protein(s) was found in the nuclear extract of Canton S embryos that specifically binds to the core P but not to the KP or various fragments of p pi 25.1. In addition, the mutant G6PD activity was found to be affected not only by the genotype, but also by cytoplasmic factors. PMID- 2560633 TI - [UV-inactivation of microorganisms in water]. AB - UV-Inactivation of Escherichia coli, Bacillus subtilis spores, Staphylococcus Phage A 994, Poliovirus type Mahoney and Rotavirus SA 11 was tested under controlled physical conditions. B. subtilis-spores were found to be the most resistant of these microorganisms, followed by Rotavirus, Bacteriophage and Poliovirus. E. coli required the lowest irradiation dose for inactivation. Causes and meaning of these dose-survival-reactions are discussed. PMID- 2560634 TI - Tubulointerstitial nephritis caused by the antiviral agent foscarnet. AB - The antiviral agent foscarnet has long been used in our unit to treat cytomegalovirus (CMV) infections in renal transplant patients. The clinical effect has been convincing and, apart from changes in serum calcium levels, very few side effects have been noted. We have, however, observed a nephrotoxic reaction in a series of patients with initially good renal function who therefore received high doses of foscarnet. Transplant biopsies performed in five of those patients revealed degenerative changes in the tubular epithelial cells as well as tubular calcium deposits and an infiltration of the interstitium by mixed mononuclear and polymorphonuclear leucocytes. Renal insufficiency was accompanied by high fever. After withdrawal of the drug, the temperature rapidly normalized, whereas serum creatinine continued to rise for about 3 days and then fell back towards previous levels. We conclude that transplant biopsies are of great value in distinguishing between a foscarnet nephrotoxic effect and CMV nephritis, various forms of rejection, and other causes of impaired renal function. PMID- 2560635 TI - Isotope transport studies and shunt pressure measurements as a guide to shunt function. AB - Shunt function testing by radionuclide injection and shunt pressure measurement was performed in 312 cases. In our experience the test has a sensitivity of 92% and accuracy of 89%. The diagnostic value of the components of the test are discussed. The test is a safe and integral part of the assessment of shunt function. The significance of shunt malfunction must of course be determined by clinical evaluation, measures of ventricular size by CAT scanning or ultrasound, and by continuous ICP recording. However the shunt function test combined with shunt pressure measurement is a safe and essential investigation of symptomatic shunt malfunction. PMID- 2560636 TI - Voltage dependent calcium channel expression in isolated osteoclasts. AB - In this study the expression of voltage-dependent calcium channels on osteoclast plasma membrane has been investigated. We found that osteoclasts were sensitive to KCl-induced depolarization. In this circumstance a 4 fold transient cytosolic calcium concentration ([Ca2+]i) increase was observed. This increase was dose dependent. Its half maximal effect was achieved at 30 mM KCl. Voltage sensitive calcium channels in osteoclasts were inhibited by specific antagonists. Nicardipine, a dihydropyridine derivative, was the most effective, inducing complete block of the channels at 10(-6) M. Verapamil (phenylalkylamine) and diltiazem (benzodiazepine) were less effective. These results are consistent with the presence, on the osteoclast membrane, of L-type voltage-sensitive calcium channels. PMID- 2560637 TI - Alterations in the expression and gating of Drosophila sodium channels by mutations in the para gene. AB - Mutations in the para gene specifically affect the expression of sodium currents in Drosophila. While 65% of wild-type embryonic neurons in culture express sodium currents, three distinct mutations in the para locus resulted in a decrease in the fraction of cells from which sodium currents could be recorded. This reduction was allele-dependent: macroscopic sodium currents were exhibited in 49% of the neurons in parats1 cultures, 35% in parats2, and only 2% in paraST76. Voltage-clamp experiments demonstrated that the parats2 mutation also affected the gating properties of sodium channels. These results provide convincing evidence that para, a gene recently shown to exhibit sequence similarity to vertebrate sodium channels alpha subunits, encodes functional sodium channels in Drosophila. The finding that one para allele (paraST76) can virtually eliminate the expression of sodium currents strongly argues that the para gene codes for the majority of sodium channels in cultured embryonic neurons. PMID- 2560638 TI - Rescue of excitation-contraction coupling in dysgenic muscle by addition of fibroblasts in vitro. AB - Muscular dysgenesis (mdg) in mice causes the failure of excitation-contraction (E C) coupling in skeletal muscle. Cultured dysgenic muscle fails to contract upon depolarization, lacks typical muscle ultrastructure, including normal triads, and lacks functional voltage-dependent slow calcium channels. We show that normal rodent fibroblasts and 3T3 fibroblasts "rescue" dysgenic myotubes, reestablishing contractions (i.e., E-C coupling), normal ultrastructure, and functional slow calcium channels. These results support the finding that the expression of the slow calcium channel is affected in the mdg mutation and that this protein is essential for E-C coupling. Additionally, fibroblast rescue provides a system for examining the mechanisms of heterotypic cellular influence on cell function. PMID- 2560639 TI - Glial and neuronal forms of the voltage-dependent sodium channel: characteristics and cell-type distribution. AB - Two functionally different forms of the voltage-dependent sodium channel were observed in glia and in neurons of the mammalian nervous system. Both forms had identical conductance and tetrodotoxin sensitivity and displayed steady-state inactivation, a strongly voltage-dependent rate of activation, and a faster but weakly voltage-sensitive rate of inactivation. However, the glial form had significantly slower kinetics and a more negative voltage dependence, suggesting that it was functionally specialized for glia. This form was found in most glial types studied, while the neuronal form was observed in retinal ganglion cells, cortical motor neurons, and O2A glial progenitor cells. Both forms occurred in type-2 astrocytes. The presence of the glial form correlated with the RAN-2 surface antigen. PMID- 2560640 TI - Developmentally regulated expression of specific tau sequences. AB - Tau protein undergoes a shift in its molecular mass and its electrophoretic complexity during early postnatal development. We have sequenced a tau cDNA from an adult rat brain expression library and have found two inserted sequences. One of these inserts predicts a fourth repeated sequence homologous to the other three in the carboxyl end of tau that have the property of microtubule binding. Oligonucleotide probes directed against the insert hybridized only to tau mRNA at postnatal time points, even though tau is first expressed as early as embryonic day 13. A probe directed against the junction revealed expression of non-insert containing tau mRNA from embryonic day 14 until postnatal day 8, after which time there was an abrupt decline in the expression of this immature form. Comparison of the developmentally expressed tau sequences with those sequences obtained directly from Alzheimer paired helical filaments revealed the presence of both the mature and the immature tau mRNA sequences. PMID- 2560641 TI - Two types of calcium channels coexist in peptide-releasing vertebrate nerve terminals. AB - The properties of the Ca2+ channels mediating transmitter release in vertebrate neurons have not yet been described with voltage-clamp techniques. Several types of voltage-dependent Ca2+ channels are known to exist on neuronal somata, but the small size and inaccessibility of most vertebrate nerve endings have precluded direct characterization of the presynaptic channels. However, large nerve endings, which release the peptides oxytocin and vasopressin in a Ca2(+) dependent manner, can be dissociated from the rat neurohypophysis. Using both single-channel and whole-cell patch-clamp techniques, we have characterized two types of Ca2+ channels that coexist in these terminals. One is a large conductance, high-threshold, dihydropyridine-sensitive channel that contributes a slowly inactivating current. The second is a smaller conductance channel, which is also activated at high thresholds, but underlies a rapidly inactivating, dihydropyridine-insensitive current. Both types of Ca2+ channels may participate in the peptide release process. PMID- 2560642 TI - Electrical activity, cAMP, and cytosolic calcium regulate mRNA encoding sodium channel alpha subunits in rat muscle cells. AB - The number of sodium channels increases sharply during development of rat skeletal muscle cells in vitro. An 8.5 kb mRNA encoding sodium channel alpha subunit rises to a peak on day 13 in vitro and falls to a value of 50% of the peak by day 18, consistent with the conclusion that mRNA abundance is a major determinant of the rapid rise in sodium channel number. Electrical activity and increased cytosolic calcium decrease the level of alpha subunit mRNA, and cAMP increases its level in parallel with changes in the number of sodium channels. The similarity between the changes in mRNA levels and sodium channel density indicates that the regulation of alpha subunit mRNA level is an important mechanism of feedback regulation of sodium channel density by electrical activity in developing rat muscle cells. PMID- 2560643 TI - Elementary properties and pharmacological sensitivities of calcium channels in mammalian peripheral neurons. AB - The major component of whole-cell Ca2+ current in differentiated, neuron-like rat pheochromocytoma (PC12) cells and sympathetic neurons is carried by dihydropyridine-insensitive, high-threshold-activated N-type Ca2+ channels. We show that these channels have unitary properties distinct from those of previously described Ca2+ channels and contribute both slowly inactivating and large sustained components of whole-cell current. The N-type Ca2+ currents are modulated by GTP binding proteins. The snail toxin omega-conotoxin reveals two pharmacological components of N-type currents, one blocked irreversibly and one inhibited reversibly. Contrary to previous reports, neuronal L-type channels are insensitive to omega-conotoxin. N-type Ca2+ channels appear to be specific for neuronal cells, since their functional expression is greatly enhanced by nerve growth factor. PMID- 2560644 TI - A synaptic vesicle membrane protein is conserved from mammals to Drosophila. AB - The structure of synaptobrevin, an intrinsic membrane protein of small synaptic vesicles from mammalian brain, was studied by purification and molecular cloning. Its message in bovine brain encodes a 116 amino acid protein whose sequence reveals it to be the mammalian homolog of Torpedo VAMP-1. Antibody probing demonstrates that the protein is also present in Drosophila, and its Drosophila homolog was cloned. Alignment of the sequences of synaptobrevin/VAMP-1 from the three species shows it to contain four domains, including a highly conserved central region of 63 amino acids that contains 75% invariant residues. The finding that a membrane protein from vertebrate synaptic vesicles is conserved in Drosophila points toward a central role of this protein in neurotransmission and should allow a genetic approach to neurotransmitter release. PMID- 2560645 TI - Formation and modulation of chemical connections: evoked acetylcholine release from growth cones and neurites of specific identified neurons. AB - The ability to release acetylcholine (ACh) from cultured neurons of Helisoma was assessed by micromanipulating ACh-sensitive somata into contact with presynaptic neurons. ACh release was reliably detected from neurites and growth cones of cholinergic neuron B5, but not neuron B19, as early as 3 s after contact with novel target neurons. The rapid onset of transmission correlates with the ability of neuron B5, but not neuron B19, to indiscriminately form chemical connections and may be related to the specificity of synaptogenesis. The neuropeptide FMRFamide reduces ACh release at early chemical connections. The rapid onset of functional transmission and the ability of FMRFamide to modulate chemical transmission at this early chemical connection suggest that neuron B5 acquires its presynaptic apparatus through an intrinsic program independently of target contact. PMID- 2560646 TI - The alpha 1 and alpha 2 polypeptides of the dihydropyridine-sensitive calcium channel differ in developmental expression and tissue distribution. AB - The dihydropyridine (DHP) binding complex isolated from skeletal muscle contains two large proteins, alpha 1 and alpha 2, and at least two smaller polypeptides. The alpha 1 subunit has a primary structure expected for ion channels and is a functional component of a DHP-sensitive, voltage-activated calcium channel. The functions of the alpha 2 protein and the smaller polypeptides are unknown. We prepared monoclonal antibodies to the alpha 1 and alpha 2 polypeptides and studied the developmental appearance and tissue distribution of these two proteins. In rat skeletal muscle, the levels of alpha 1 are quite low during the first 10 days after birth, then rise dramatically, and, by day 20, approach those found in adult muscle. In direct contrast, alpha 2 is present in substantial amounts in rat muscle at birth and increases only slightly during this same period of development. Furthermore, alpha 1 is detected only in skeletal muscle, whereas alpha 2 is present in a variety of tissues. These results provide evidence for the segregation of these two polypeptides and suggest that the function of alpha 2 is not limited to that associated with the DHP-sensitive calcium channel. PMID- 2560647 TI - Calcitonin gene-related peptide regulates phosphorylation of the nicotinic acetylcholine receptor in rat myotubes. AB - The nicotinic acetylcholine receptor (AChR) is a substrate for at least three different protein kinases, and phosphorylation of the receptor has been shown to increase its rate of desensitization. However, the first messengers that regulate AChR phosphorylation have not yet been identified. This study demonstrates that calcitonin gene-related peptide (CGRP), a neuropeptide present in the axon terminals of the neuromuscular junction, regulates phosphorylation of the AChR in primary rat myotube cultures. CGRP, in the presence of the phosphodiesterase inhibitor Ro 20-1724, increased phosphorylation of the alpha and delta subunits of the AChR. CGRP-induced phosphorylation of the AChR had the same subunit specificity and temporal sequence as previously observed using forskolin or cAMP analogs. Phosphorylation of the AChR in the presence of CGRP appears to be mediated by CGRP-stimulated increases in cAMP levels leading to activation of cAMP-dependent protein kinase. The present results, taken together with the recent demonstration that CGRP increases the rate of AChR desensitization in mouse myotubes, suggest that CGRP may play a physiological role as a regulator of AChR desensitization by modulating AChR phosphorylation at the neuromuscular junction. PMID- 2560648 TI - NGF induction of the gene encoding the protease transin accompanies neuronal differentiation in PC12 cells. AB - Various proteases have been found to be released by the growth cones of developing neurons in culture and have been hypothesized to play a role in the process of axon elongation. We report here that nerve growth factor (NGF) induced the gene encoding the metalloprotease transin in PC12 cells with a time course coincident with the initial appearance of neurites by these cells. Acidic and basic fibroblast growth factors also stimulated transin mRNA expression and neurite outgrowth, whereas various other agents had no effects on either of these phenomena. In contrast, dexamethasone was found to inhibit the induction of transin mRNA when added with, or following, NGF treatment. Finally, we show that sequences contained within 750 bp of the 5' untranscribed region of the transin gene confer responsiveness to NGF and dexamethasone. PMID- 2560649 TI - Expression of nerve growth factor receptor mRNA is developmentally regulated and increased after axotomy in rat spinal cord motoneurons. AB - In situ hybridization histochemistry and RNA blot analysis were used to study expression of nerve growth factor receptor (NGF-R) mRNA in rat spinal cord motoneurons. The results show that NGF-R mRNA is expressed at high levels in rat spinal cord motoneurons at the time of naturally occurring cell death. This expression is sustained, but reduced, during synapse formation and is subsequently greatly reduced in the adult spinal cord. A unilateral crush lesion of the sciatic nerve resulted in an 8-fold increase in NGF-R mRNA in adult rat spinal cord motoneurons 3 days after lesion, compared with the nonlesioned side. NGF-R mRNA induction was even more pronounced 7 and 14 days after lesion, reaching levels 12 times higher than those on the nonlesioned side. However, 6 weeks after lesion, when the motor function of the leg was largely restored, NGF R expression had decreased to levels similar to those on the contralateral side. We therefore suggest that NGF-R mediates a trophic or axonal guidance function for developing and regenerating spinal cord motoneurons. PMID- 2560650 TI - Development alters the expression of calcium currents in chick limb motoneurons. AB - Calcium current types expressed in vertebrate spinal motoneurons have not been previously resolved. We have resolved three types in chick limb motoneurons identified by retrograde labeling and report that dramatic changes in their expression take place during development in vivo. T-, N-, and L-type calcium currents were distinguished on the basis of kinetics, voltage dependence, and unique pharmacological sensitivities. Developmental changes were characterized by studying motoneurons isolated from embryos at three stages spanning neuromuscular system development. T currents were dominant at the earliest stage. Motoneurons from embryos 2 days older showed much reduced T currents and much increased N and L currents. We suggest that mature motoneurons will be dominated by N- and L-type calcium currents and that T current may serve developmental roles. PMID- 2560651 TI - Effects of ciclosporin and protein synthesis inhibitors on cutaneous inflammation in mouse skin. AB - Ciclosporin is clinically effective in a variety of inflammatory skin diseases. We have therefore studied the effects of the drug on cutaneous inflammation in mice. Ciclosporin inhibited the inflammatory response to 12-O tetradecanoylphorbol-13-acetate (TPA) and to the contact sensitising agent oxazolone when applied topically to mouse skin. The drug had no effect on arachidonic acid-induced inflammation. The protein synthesis inhibitor cycloheximide showed a similar profile of activity. Ciclosporin, like actinomycin D but unlike cycloheximide, was only effective in inhibiting the inflammatory response to TPA if given 0.5 h before, but not 2 h, after TPA. These results suggest that the anti-inflammatory activity of ciclosporin in the skin is due to an effect on the production of proinflammatory proteins. PMID- 2560652 TI - Leukotriene B4 receptors on neutrophils in patients with psoriasis and atopic eczema. AB - Polymorphonuclear leukocyte (PMNL) infiltration is an important characteristic in psoriatic lesions. Elevated concentrations of the chemoattractant eicosanoid leukotriene B4 (LTB4) are present in psoriatic skin. Its chemotactic activity is mediated via high affinity receptors on PMNL. The goal of our work was to ascertain whether PMNL infiltration in psoriasis can be accounted for by functional abnormalities of the circulating PMNL due to alterations in the LTB4 receptor density or affinity (or both). No significant difference was found between patients with psoriasis, healthy controls and patients with another inflammatory dermatosis (atopic eczema) with regard to the binding parameters of LTB4 receptors on PMNL. Our findings suggest that PMNL accumulation in psoriatic skin may be the result of an excess of cutaneous chemoattractant rather than the increased readiness of psoriatic PMNL to migrate towards LTB4 due to altered LTB4 receptor density or affinity. PMID- 2560653 TI - The nuclear envelope. PMID- 2560654 TI - Chromosomes and chromatin structure: the extrachromosomal karyoskeleton. PMID- 2560655 TI - Regulation of gene expression by nuclear hormone receptors. PMID- 2560656 TI - DNA topoisomerases. PMID- 2560657 TI - Strategies for identifying functionally active domains upon cell surface receptors. PMID- 2560659 TI - The molecular constituents of intercellular junctions. PMID- 2560658 TI - Antiporters. PMID- 2560660 TI - Cell-to-cell interactions during synaptogenesis. PMID- 2560661 TI - Collagens and elastin: structure and interactions. PMID- 2560662 TI - Concentration-dependent, stereoselective inhibition of the endotoxin-induced hemoconcentration in conscious rats with the peptidoleukotriene receptor antagonist SK & F 104353. AB - Endotoxemia is associated with increases in a number of humoral mediators including vasopressin, thromboxane and leukotrienes (LT), all of which may participate in the pathophysiologic responses to endotoxemia. Previous studies from our laboratory demonstrated that endotoxin-induced hemoconcentration was attenuated with a peptidoleukotriene receptor antagonist, SK & F 104353. The purpose of this study was to investigate further the mechanism of endotoxin induced hemoconcentration. Injection of LTD4 (51 nmol/kg, i.v.) produced an increase in the hematocrit of conscious male Sprague-Dawley rats: administration of SK & F 104353 (2 mg/kg, i.v. + 10 mg/kg/h, i.v. infusion) blocked completely this response to exogenous LTD4. Injection of Salmonella enteritidis endotoxin (30 mg/kg, i.v.) increased the hematocrit from 41 +/- 1 vol% to 55 +/- 1 vol%. Following pretreatment with SK & F 104353 (2 mg/kg, i.v. + 10 mg/kg/h, i.v.), the hemoconcentration was attenuated to 46 +/- 1 vol% (p less than 0.01). Simultaneous determination of plasma drug concentrations over a range of doses indicated that inhibition of the hemoconcentration produced by SK & F 104353 was concentration-dependent (IC30 = 0.5 microgram/ml). The IC30 for the stereoisomer, SK & F 104373, was 50 micrograms/ml. The 5-lipoxygenase/cyclooxygenase inhibitors, SK & F 86002 and BW 775C, also attenuated the endotoxin-induced increase in hematocrit, whereas indomethacin, heparin, daltroban, or the selective V1 vasopressin receptor antagonist [d(CH2)5Tyr(Me)]AVP did not significantly affect the endotoxin-induced hemoconcentration. The endotoxin induced hemoconcentration was inhibited in a concentration-dependent, stereoselective manner with a peptidoleukotriene receptor antagonist, and by 5 lipoxygenase inhibitors, indicating that this response is mediated by peptidoleukotrienes. PMID- 2560664 TI - Restriction endonuclease analysis of four Borrelia burgdorferi strains. AB - A restriction endonuclease analysis was performed on four strains of Borrelia burgdorferi: one isolated from man (SF), one from Ixodes dammini (B31) and two from I. ricinus (BITS in Italy and B45 in Germany). Digestion by Taq I and Hae III gave the best resolution of the DNA fragments. Three different restriction patterns were obtained: BITS and B45 showed only one band difference. These results correlate with the reactivity of the four strains with monoclonal antibodies. PMID- 2560665 TI - Magnetic resonance imaging of liver tumors. AB - Careful optimization of scanning techniques, particularly motion artifact suppression, has been essential to achieve reproducible results in abdominal MRI. The investigators experience indicates that MRI can be more accurate than other imaging methods for the detection of focal liver lesions. Furthermore, MRI is able to solve the major clinical problems in differential diagnosis of benign and malignant liver lesions: cancer v cavernous hemangioma or focal fat. MRI has reduced the dependence on liver biopsy and angiography to diagnose and stage focal liver lesions. Unfortunately, both imaging techniques, especially motion artifact suppression methods, vary widely among machines operating at different field strengths. Therefore, as hardware and software evolve, it is necessary to retrace the steps of pulse sequence optimization and clinical testing. Hopefully, in the future, standardized imaging techniques will become available for body MRI. PMID- 2560663 TI - Rabbit leukocytes activated by C5a des Arg and N-formyl-methionyl-leucyl phenylalanine promote endothelial prostacyclin production. Evidence for a role of hydrogen peroxide. AB - Prostacyclin (PGI2) production by an intact endothelial cell layer on the surface of isolated rabbit aortas was investigated in the presence of chemoattractants (C5a des Arg, FMLP, LTB4, PAF) and autologous blood leukocytes. Leukocytes themselves did not produce detectable amounts of PGI2, nor did they alter basal endothelial PGI2 formation. In the absence of leukocytes, C5a des Arg (but not FMLP, LTB4 or PAF) produced a direct and dose-dependent stimulation of PGI2 release from endothelial cells. This effect was greatly enhanced when C5a des Arg and leukocytes were incubated together on the endothelium. Similar results were obtained on co-incubation of endothelium and leukocytes with FMLP and to a lesser extent with LTB4 and PAF. Experiments with superoxide dismutase and catalase suggest that release of H2O2 may partly explain the stimulatory activity of leukocytes in the presence of chemoattractants. The effect of aspirin treatment of the endothelial cells in enhancing leukocyte adherence induced by C5a des Arg suggests that local PGI2 production by vascular endothelium may provide a natural defence mechanism by attenuating cell-cell interactions. PMID- 2560666 TI - [Cancer of the rhinopharynx (clinical picture, scanning and statistics)]. PMID- 2560667 TI - [Recent progress in endothelin research]. PMID- 2560668 TI - [Simultaneous in situ detection of viral antigens and RNA in brain tissues from a patient with herpes simplex encephalitis by immunocytochemistry and in situ hybridization]. AB - In order to elucidate the correlation between herpes simplex virus (HSV-1) and the central nervous system tissue, we performed the simultaneous detection of viral antigens and RNA in the brain tissue sections from a patient with herpes simplex virus (HSV) encephalitis using immunocytochemistry and in situ hybridization. In the present study the hybridization protocol reported by Brahic M et al. in 1984 were applied for the simultaneous detection of viral RNA and antigens with a few modification. The sections were first immunocytochemically stained to detect HSV-1 antigens by ABC method, and then hybridized with 3H labelled HSV-1 cDNA probe for the detection of RNA after the acetylation of slides for the prevention of nonspecific bindings of isotope to slides. In the present study, viral antigens were immunocytochemically stained to brown-colored deposits located in the cytoplasm and nucleus whereas viral RNA were detected as the accumulation of many silver grains over the nuclei or cytoplasm. In this case the light microscopic findings in a part of temporal lobe showed multiple areas of necrosis mainly involving the gray matter and a few inflammatory changes such as perivascular cell cuffings. HSV-1 infected Vero cells as positive control demonstrated both antigens and RNA as shown in Fig.1 a. However, no hybridization signals and color deposits were observed in uninfected Vero cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2560670 TI - Tuberculosis control in the Ivory Coast: a brief synopsis, March 1989. PMID- 2560669 TI - [The role of single-stranded DNA in the integration of SV40 genome into cellular DNA]. AB - The integration of temperature-sensitive SV40 mutant DNA (tsA239) into the Chinese hamster cellular genome at an early stage of infection was studied. The content of single-stranded DNA structures in the infected and control cells at a non-permissive temperature (40 degrees C) differed drastically from that in control cells at permissive temperatures (33 degrees C, 37 degrees C). The role of single-stranded structures in the integration of the SV40 genome into cellular DNA was shown by blot hybridization. The integration mechanism is discussed. PMID- 2560671 TI - Evaluation of tuberculosis prevalence and of the efficiency of case-finding in several bio-climatic zones of the Ivory Coast. PMID- 2560672 TI - [Sodium conductin of the human brain: purification and functional characterization]. AB - A sodium channel enriched preparation was obtained from human brain. Human sodium channel appeared as a heterocomplex peptide alpha beta 1 beta 2. Functional properties of the protein were maintained since, after reconstitution into liposomes, ion fluxes were sensitive to sodium channel specific toxins and to membrane potential. Moreover, the reconstituted protein showed a well defined ionic selectivity. PMID- 2560673 TI - A two-stage carcinogenesis model for risk assessment. PMID- 2560674 TI - Effects of aging on alpha 1-adrenoceptor mechanisms and the inhibitory effect of diltiazem on noradrenaline maximum response in isolated rat aortic preparation. AB - The effects of aging on alpha 1-adrenoceptor mechanisms in aortic preparations isolated from 3-, 6-, 10-, 18-, and 40-week-old rats were studied and compared with serotonin receptor mechanisms in the same preparations. The potency (pD2 value) of noradrenaline increased with age from 3 to 10 weeks, but decreased thereafter with age from 10 to 40 weeks. The affinity (pKA value) of noradrenaline and of prazosin (pA2 value) did not alter with aging. The change in potency or the pD2 value of noradrenaline was proportional to receptor reserve (pD2-pKA value) for noradrenaline, suggesting that the change of potency of noradrenaline with age was due to a change of receptor reserve, but not to change of drug affinity to alpha 1-adrenoceptors. The potency (pD2 value) and affinity (pKA value) of serotonin, and the affinity (pA2 value) of ketanserin, did not alter with aging, suggesting that serotonin receptor mechanisms in rat aorta did not change with age. The inhibitory effect of diltiazem on noradrenaline maximum response decreased with age from 3 to 10 weeks, but increased with age from 10 to 40 weeks. An inverse relationship between changes of diltiazem inhibition and receptor reserve of noradrenaline was found. Diltiazem's inhibitory effect on serotonin maximum response did not alter with aging. PMID- 2560675 TI - The effect of sulfonylurea therapy on defective calcium movement associated with diabetic cardiomyopathy. AB - Adult rats exposed to 70 mg/kg streptozocin developed characteristic symptoms of overt diabetes, such as muscle wasting and severely elevated blood glucose levels. Chronic treatment of these rats with the sulfonylurea glyburide for a period of 5 weeks did not affect either the weight of the animal or the degree of hyperglycemia. The drug also failed to influence myocardial glucose metabolism. Nevertheless, the decline in myocardial function associated with the diabetic cardiomyopathy was less in the glyburide-treated rats. At higher preload, myocardial work was significantly reduced in the untreated diabetic but was only moderately depressed in the glyburide-treated heart relative to the nondiabetic heart. The improvement in mechanical function was associated with partial recovery of sarcolemmal calcium pump activity. The drug did not alter the initial rate of Na+-Ca2+ exchange, but decreased the capacity of the transport system. The results indicate that glyburide benefits the diabetic heart by a mechanism independent of carbohydrate metabolism. PMID- 2560676 TI - Species differences in the positive inotropic response to DPI 201-106, a novel cardiotonic agent. AB - The positive inotropic activity of the novel cardiotonic DPI 201-106 was investigated in rat and guinea pig isolated hearts. For comparative purposes, the adenylate cyclase stimulant forskolin and the sodium channel agonist veratridine were also evaluated in both species. DPI 201-106 and veratridine produced greater inotropic effects in rat hearts than in guinea pig hearts, whereas forskolin produced comparable effects. In both species the inotropic response to DPI 201 106 and veratridine, but not forskolin, was reversed by the sodium channel antagonist tetrodotoxin. These results confirm that the positive inotropic effect of DPI 201-106 is due to stimulation of the sodium channel and demonstrate for the first time that species differences exist in the inotropic response to this novel cardiotonic drug. PMID- 2560677 TI - Possible role of Na(+)-Ca2+ exchange in the regulation of contractility in isolated adult ventricular myocytes from rat and guinea pig. AB - Action potentials and developed contractions of externally unloaded single ventricular myocytes isolated from adult rat and guinea pig hearts were recorded by means of an optical system for recording contractile activity during regular stimulation by microelectrodes. Under control conditions, the shortenings (twitches) in the rat myocytes were fully inhibited by 0.1 microM ryanodine, but they were rather insensitive to the Ca2+ blocker 0.2-0.5 microM nifedipine. In contrast, the contractions of the isolated guinea pig ventricular myocytes were greatly suppressed by 0.2-0.5 microM nifedipine (to less than 30%), while they were only slightly reduced by 1 microM ryanodine. When the Na+ gradient was decreased by reducing [Na]o or by elevating [Na]i in the presence of veratridine, the twitch contractions were increased in both species. The effect of reduced [Na]o on twitch contractions was not affected by ryanodine in either type of myocytes, while nifedipine still fully abolished the twitches in the guinea pig cells, indicating a strong dependence of guinea pig contractions on Ca2+ influx. On the other hand, the effect of a reduced Na gradient by veratridine was more complex; the usual twitch (phasic component) was increased and it was followed by a second (tonic) component which relaxed only after the repolarization of the action potential. While the phasic component was decreased by nifedipine and ryanodine in the usual way (as in the controls), the sustained contractions (lasting up to several seconds) were ryanodine and nifedipine insensitive. Furthermore, the cardiomyocytes of both species exposed to strontium in place of external calcium still exhibited all the effects observed when reducing the Na+ gradient.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2560678 TI - Selective inhibition of [3H]nitrendipine binding to brain and cardiac membranes by bacitracin. AB - The effects of bacitracin were investigated on [3H]nitrendipine binding to rat brain and cardiac membranes in a low ionic strength (5 mM Tris-HCl) buffer. Bacitracin inhibited [3H]nitrendipine binding to rat brain and cardiac membranes with IC50 values of 400 +/- 100 and 4600 +/- 400 micrograms/mL, respectively. Scatchard analysis in brain membranes revealed that bacitracin inhibited [3H]nitrendipine binding primarily by reducing the Bmax but also by producing a small increase in the Kd. In brain membranes, Na+ (100 mM) and Ca2+ (2 mM) reduced the potency of bacitracin to inhibit [3H]nitrendipine binding by approximately sixfold with IC50 values of 2600 +/- 300 and 2100 +/- 400 micrograms/mL observed for bacitracin in the presence of 100 mM Na+ and 2 mM Ca2+, respectively. The EC50 values for the effects of Na+ and Ca2+ were 800 +/- 200 microM and 25 +/- 5 mM. K+, Mg2+, choline, and increasing the assay buffer of Tris-HCl to 50 mM also decreased the inhibition of [3H]nitrendipine binding by bacitracin. These results suggest that bacitracin specifically modulates [3H]nitrendipine binding in a cation-dependent manner and that brain and cardiac dihydropyridine binding sites are either biochemically different or exist in a different membrane environment. PMID- 2560680 TI - Dual-fluorescence flow cytometric analysis of membrane potential and cytoplasmic free Ca2+ concentration in embryonic rat hippocampal cells. AB - We have demonstrated simultaneous measurement of the membrane potential and cytoplasmic free Ca2+ concentration ([Ca2+]i) by utilizing a dual-laser flow cytometer in embryonic rat hippocampal cell suspensions. Veratrine, a Na+ channel activator, induced both membrane depolarization and elevation of [Ca2+]i. These actions of veratrine were all reversed by the presence of tetrodotoxin (TTX). These findings suggest that Na+ channels are functionally expressed in the cells and the activation of Na+ channels increases [Ca2+]i. The usefulness of the flow cytometric analysis in elucidating the expression of membrane functions in the embryonic central nervous systems (CNS) is discussed. PMID- 2560679 TI - Pregnancy-related changes of galactose recognition system on sinusoidal rat liver cells. AB - We have studied the galactose specific receptor activities of sinusoidal liver cells of pregnant rat from the 13th day of gestation to the 10th day after delivery. Both liver macrophages and endothelial cells modulate the expression of hepatic binding protein during rat pregnancy and lactation. Except for endothelial cells of rats at the 22nd day of gestation, we have measured an overall decrease of binding sites. In addition, in liver macrophages the clustered distribution of galactose specific receptors, which is typical for adult animals, is reduced or completely absent. These results indicate that the galactose specific receptors on non-parenchymal liver cells are modulated in relation with the physiological state of the organ like the asialoglycoprotein receptor of hepatocytes. PMID- 2560681 TI - Synthesis and function of the mitochondrial intron--encoded bI4 RNA maturase from Saccharomyces cerevisiae. Effects of upstream frame-shift mutations. AB - We have analyzed the expression and function of the intron-encoded bI4 maturase when frame-shift mutations in the upstream exon alter the translational process. By constructing secondary cis-acting mutations within the bI4 intron, we observed (1) that the bI4 maturase is still translated in the presence of the upstream mutation, albeit in very low amounts, and (2) that the limited amounts of bI4 maturase made under these conditions is no longer able to promote the splicing process of the aI4 intron. These observations, which further strengthen the maturase model, strongly suggest that bI4 maturase acts sequentially on the bI4 intron and then on the aI4 intron. PMID- 2560682 TI - A hot-spot for transposition of various Ty elements on chromosome V in Saccharomyces cerevisiae. AB - Ty4 is a novel transposable element in the yeast, Saccharomyces cerevisiae, which is present in only a few copies in the genome (Stucka et al. 1989). In strain C836 one of the three copies (Ty4-90) is contained in cosmid clone c90, where it resides on chromosome V. Analysis of this region reveals a "hot-spot" of transposition: in addition to Ty4-90, the locus contains a complete Ty3 element and seven singular delta, sigma and tau elements. Three tRNA genes (for His, Lys, and Ile) are located in this region, and these are closely associated with one or the other of the elements, a phenomenon commonly observed in yeast. A comparison of c90 with corresponding regions from other strains shows that the locus is highly polymorphic and that this polymorphism is explicitly associated with Ty transposition and recombination events. PMID- 2560684 TI - Comparison of the ACTH and cortisol responses to provocative testing with glucagon and insulin hypoglycaemia in normal subjects. AB - The glucagon stimulation test (GST) is often used to assess pituitary ACTH reserve, particularly when other tests are contra-indicated. In a preliminary investigation, in patients with pituitary disease, we failed to demonstrate the ACTH dependence of the cortisol response. We have therefore compared the ACTH, cortisol and glucose responses to glucagon (1 mg s.c.), insulin (0.2 U/kg i.v., ITT) and placebo in six healthy male volunteers, sampling every 10 min for 6 h. During the GST, mean +/- SD serum cortisol rose from 256 +/- 80 nmol/l to a peak of 481 +/- 164 nmol/l (range 289-717 nmol/l, P less than 0.01) in comparison with 280 +/- 81 nmol/l to 602 +/- 110 nmol/l (range 493-742 nmol/l) during the ITT (P less than 0.002). The mean peak cortisol levels achieved in the two tests did not differ significantly. In the GST, plasma ACTH rose from a mean basal value of 10.9 +/- 16.6 ng/l to a mean peak level of 123 +/- 76 ng/l (P less than 0.02) (ACTH ng/l x 0.225 = pmol/l). The corresponding values in the ITT were 7.1 +/- 16.2 ng/l and 263 +/- 91 ng/l (P less than 0.001). The mean peak ACTH level was significantly greater during the ITT (P less than 0.05). Thus the cortisol response was ACTH dependent in both the GST and the ITT in normal subjects. Furthermore, the ACTH response was of sufficient duration to be detected by the usual procedure of sampling every 30 min.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2560683 TI - Cytochrome oxidase subunit II sequences in Petunia mitochondria: two intron containing genes and an intron-less pseudogene associated with cytoplasmic male sterility. AB - The mitochondrial genome of Petunia hybrida contains two transcribed cytochrome oxidase subunit II (coxII) genes. The coding region of both genes is split by a 1.3 kb group II intron. Unlike coxII-1, which is similar to other sequenced plant coxII genes, the coxII-2 coding region is extended by 48 codons. The cytoplasmic male sterile (CMS) Petunia contains one coxII gene similar in structure and transcript pattern to the coxII-1 gene found in the fertile genome. Comparison of the sequenced coxII genes from the fertile mitochondrial genome with the coxII sequences present in the CMS-associated pcf gene from the CMS genome (Young and Hanson 1987) suggests that pcf is a processed pseudogene. A model for the generation of pcf is presented. PMID- 2560685 TI - Effect of porcine brain natriuretic peptide (pBNP) on human adrenocortical steroidogenesis. AB - Porcine brain natriuretic peptide (pBNP), purified from porcine brain, had a significant suppressive effect on aldosterone and cortisol secretions in ACTH treated cultured human adrenal cells. Concomitantly, the intracellular cGMP formation was enhanced by pBNP treatment. A specific pBNP receptor was identified in the human adrenal tissues. Affinity labelling of 125I-pBNP showed two separate molecular weights of specific binding sites for pBNP of 140 and 67 kDa. A 125I pBNP binding study of the human adrenal membrane fraction demonstrated the presence of high-affinity and low-capacity binding sites for pBNP. Moreover, these binding sites for 125I-pBNP were displaced by unlabelled alpha-rANP as well as pBNP. From these studies, we concluded that pBNP had suppressive effects on human adrenocortical steroidogensis, possibly via a receptor which may be shared with ANP. PMID- 2560686 TI - Effects of inhibition of cholesterol synthesis by simvastatin on the production of adrenocortical steroid hormones and ACTH. AB - Simvastatin, a derivative of lovastatin, is a potent inhibitor of cholesterol biosynthesis and may interfere with steroid hormone production, for which cholesterol is required. In a single-blind, placebo-controlled study, 24 patients with severe primary hypercholesterolaemia (mean serum cholesterol +/- SD = 10.74 +/- 1.59 mmol/l) were treated with simvastatin 40 mg per day for 8 weeks. Before and after treatment, the following parameters were evaluated: basal levels of ACTH, cortisol, androstenedione, dehydroepiandrosterone and 17 hydroxyprogesterone; urinary excretion of free cortisol; the cortisol response after short-term infusion of ACTH; the ACTH and cortisol response during insulin induced hypoglycaemia. Total serum cholesterol decreased by 35.0 +/- 8.1% (P less than 0.001) and low-density lipoprotein (LDL) cholesterol by 39.8 +/- 9.8% (P less than 0.001); high-density lipoprotein (HDL) increased by 9.2 +/- 11.1% (P less than 0.001). Basal levels of ACTH were higher after simvastatin (2.9 +/- 1.9 pmol/l vs 4.1 +/- 2.9 pmol/l; P less than 0.05) whereas basal levels of steroid hormones were not significantly changed. The excretion of free cortisol was unaltered. The peak cortisol after ACTH infusion was lower after treatment (0.87 +/- 0.23 mumol/l vs 0.78 +/- 0.10 mumol/l; P less than 0.05), but was unaltered during insulin-induced hypoglycaemia. We conclude that simvastatin lowers serum cholesterol without clinically relevant effects on the adrenocortical steroid hormone secretion and the hypothalamic-pituitary-adrenal axis. PMID- 2560687 TI - Corticotrophin-like intermediary lobe peptide as a marker of alternate pro opiomelanocortin processing in ACTH-producing non-pituitary tumours. AB - In order to evaluate which of human (h) corticotrophin-like intermediary lobe peptide (CLIP) or h beta-melanocyte stimulating hormone5-22 (h beta MSH5-22) was the better marker of alternate pro-opiomelanocortin (POMC) processing, both peptides were simultaneously sought in the same tissue extracts from a normal human pituitary, six corticotrophic adenomas, and four non-pituitary tumours responsible for an ectopic ACTH syndrome. Human CLIP was detected using a combination of gel exclusion chromatography and two different radioimmunoassays (RIAs): a mid-ACTH RIA which recognized ACTH but not CLIP, and a COOH-ACTH RIA which recognized both molecules. Human beta MSH5-22 had been measured previously. Neither hCLIP nor h beta MSH5-22 were detected in the normal or tumoural pituitaries. The four non-pituitary tumours, in contrast, contained both peptides; the hCLIP and h beta MSH5-22 ratios (CLIP/CLIP + ACTH and h beta MSH5 22/h beta MSH5-22 + h gamma LPH) ranged from 40 to 94% and from 24 to 46%, respectively. In a given tissue the hCLIP ratio was always higher than the h beta MSH5-22 ratio. hCLIP is therefore the better marker of alternate POMC processing. PMID- 2560688 TI - Effect of alcohol on lipoprotein metabolism. I. High density lipoprotein binding. AB - The effects of ethanol upon the binding of [125I]-labelled human high density lipoprotein 3 (HDL3) was examined in rat liver microsomes and monolayer cultures of human hepatoma (Hep G2) cells. Alcohol feeding to rats (35% caloric content) caused a significant (p less than 0.05) increase in serum cholesterol concentrations relative to pair-fed controls, but HDL3 binding to rat liver microsomes was unaffected by alcohol consumption. By contrast, addition of 10 mM ethanol to Hep G2 cells increased HDL3 binding, and this increase was observed after 14, 28 and 40 days of exposure. This alcohol-dependent rise in HDL3 binding was associated with a 2.3- to 5-fold rise in receptor number (Bmax), and a 2- to 6-fold increase in the dissociation constant (Kd). The data suggest that the net effect of increased receptor number and lower receptor affinity is to increase the capacity of hepatocytes to metabolize circulating high density lipoproteins, and that this increase in the face of elevated plasma high density lipoprotein cholesterol consequent upon alcohol consumption would facilitate greater mobilization of cholesterol from peripheral tissues to the liver. PMID- 2560689 TI - Effects of free radicals and oxidants on myocardial cellular injury. AB - Harmful effects of some biochemically generated free radicals and oxidants, including superoxide anions (O2-), hydroxyl radical (OH.), hydrogen peroxide, hypochlorous acid, and hypohalite radical, on isolated cardiac myocytes were compared in an attempt to identify the exact nature of the free radicals/oxidants responsible for myocardial ischemic-reperfusion injury. All of these free radicals/oxidants, with the exception of O2-, caused significant injury to the myocytes as evidenced by the enhanced lactate dehydrogenase and creatine kinase release as well as by morphologic examinations, simultaneously causing lipid peroxidation and oxidized glutathione release from the cells, OH. being the most detrimental of all. PMID- 2560690 TI - Serum angiotensin converting enzyme activity in type 2 (non-insulin-dependent) diabetic patients with chronic glomerulonephritis. AB - To determine the value of measuring serum angiotensin converting enzyme (ACE) activity in type 2 (non-insulin-dependent) diabetic patients who have proteinuria, 19 diabetic patients with chronic glomerulonephritis (CGN), 20 diabetic patients with diabetic glomerulosclerosis and 20 healthy controls were studied. Serum ACE activity was significantly (p less than 0.01) higher in patients with diabetic glomerulosclerosis (26.7 +/- 4.9), while in patients with CGN it was comparable to that in healthy controls (21.5 +/- 3.5, 19.1 +/- 4.7, respectively). These results suggest that measurement of serum ACE activity may be useful in differentiating between diabetic glomerulosclerosis and CGN in type 2 diabetic patients who have proteinuria. PMID- 2560691 TI - Natural history and treatment of diabetic nephropathy. PMID- 2560692 TI - Characterization of unique ADTN-catecholamine binding sites in the iris root ciliary body of rabbits. AB - A binding site for tritiated 2-amino-6, 7-dihydroxy-1, 2,3,4 tetrahydronaphthalene (ADTN) has been partially characterized in the rabbit iris root-ciliary body. Binding of ADTN is proportional to protein content and requires at least 60 minutes to reach equilibrium. Binding is saturable, with a Kd of 27 +/- 1 nM and a Bmax of 2.1 +/- .3 pmol/mg protein (mean +/- SEM). Dopamine competes for this site with a Ki of 100 nM and apomorphine with a Ki of 180 nM. This site is not blocked by L-timolol, phenoxybenzamine, or by several DA1 and DA2 antagonists. It appears to be a new type of catecholamine binding site, of a type not observed outside the anterior eye. It is possible that some of the effects of dopamine on intraocular pressure are mediated through this binding site. PMID- 2560693 TI - Cryogenic induced ocular HSV-1 reactivation is enhanced by an inhibitor of the lipoxygenase pathway. AB - Reactivation of latent herpes simplex virus type 1 (HSV-1) has been shown to occur in response to localized inflammation. Prostaglandins and lipoxygenase products [eg. hydroxyeicosatetraenoic acids, (HETEs)] are associated with inflammation and, therefore, may play a role in HSV-1 infection and reactivation. In the rabbit cornea, alkali injury, cryogenic injury, and acute HSV-1 infection promote the synthesis of HETEs. Recently, a platelet activating factor antagonist, ginkgolide B (BN 52021) has been found to specifically inhibit the corneal synthesis of HETEs after alkali injury. If the induction of HETEs after injury is related to HSV reactivation and severity of infection, BN 52021 may alter HSV reactivation and the severity of infection by reducing the production of HETEs. To study the effect of BN 52021 on HSV-1 reactivation, cryogenic corneal lesions were produced in ten HSV-1 latently infected rabbits. Five rabbits were treated with topical and intravenous BN 52021 while the remaining five rabbits received topical artificial tears and intravenous saline. In the BN 52021 treated group, 90% (9/10) of the eyes and 53% (35/66) of the total ocular cultures were positive for HSV-1. In the control group, 60% (6/10) of the eyes, and 27% (18/66) of the ocular swabs were positive for HSV-1. The total number of positive cultures was significantly greater (p less than .05) in the BN 52021 treated rabbits. By increasing the number of positive HSV ocular cultures, BN 52021 appeared to act similarly to other inhibitors of arachidonic acid metabolism such as steroidal and nonsteroidal anti-inflammatory agents. PMID- 2560694 TI - Penetrating keratoplasty in rabbits induces latent HSV-1 reactivation when corticosteroids are used. AB - The increased incidence of corneal graft failure in patients with herpes simplex virus (HSV) keratitis may be due in part to reactivation of latent HSV following surgical corneal trauma and postoperative corticosteroid therapy. To determine the onset, frequency, and nature of HSV recurrences following penetrating keratoplasty (PKP), 21 HSV type 1 (HSV-1) latently infected rabbits underwent unilateral autograft PKP. Opposite unoperated eyes served as HSV-1 latently infected controls. Corneal autografts were performed so that immunologic graft rejection would not be confused with recurrent HSV-1 stromal disease. After PKP, 11 of the 21 eyes were treated with dexamethasone. Ocular cultures and slit-lamp examinations were performed daily for the first postoperative 8 days and every other day thereafter for 82 days. Nine (82%) of the 11 dexamethasone-treated PKP eyes, 2 (20%) of the PKP eyes not treated with dexamethasone, and 3 (17%) of the 18 unoperated eyes had positive HSV-1 ocular cultures. Geographic ulcers appeared only in the PKP eyes treated with dexamethasone; 9 (82%) of the 11 PKP eyes treated with dexamethasone developed geographic ulcers. Between the 24th and 90th postoperative days, stromal keratitis appeared in 5 (56%) of the 9 PKP eyes treated with dexamethasone and in 2 (25%) of the 8 PKP eyes not treated with dexamethasone. Autograft PKP with postoperative corticosteroids significantly increased HSV-1 ocular shedding, epithelial ulceration, and stromal keratitis. This experimental model provides a useful tool to further investigate the development and treatment of HSV-1 epithelial and stromal recurrences after PKP. PMID- 2560695 TI - The Callimico goeldii (Primates, Platyrrhini) genome: karyology and middle repetitive (LINE-1) DNA sequences. AB - Callimico goeldii (Goeldi's marmoset) is a neotropical primate with 2n = 47,X1X2Y in the male, and 2n = 48,X1X1X2X2 in the female, due to a Y-autosome translocation. Karyological comparisons of Callimico, Callithrix jacchus and Cebus apella suggest that Callimico is a member of the Callitrichidae. Isozyme data and restriction mapping of LINE-1 repetitive elements in these species and in a variety of other neotropical primates confirm these findings and supply strong evidence for including Callimico in the Callitrichidae. PMID- 2560696 TI - Genomic distribution of copia-like transposable elements in somatic tissues and during development of Drosophila melanogaster. AB - The genomic distribution of elements of the copia, 412, B 104, mdg 1, mdg 4 and 1731 transposon families was compared by the Southern technique in DNA preparations extracted from brains, salivary glands and adult flies of two related Drosophila lines. The copia, 412 and mdg 1 sequences were also probed in DNA from sperm, embryos, and 1st and 2nd instar larvae. The homogeneity of the patterns observed shows that somatic transposition is unlikely to occur frequently. A correlation between mobility and the euchromatic or heterochromatic location of transposable elements is discussed. In addition, an explanation of the variable band intensities of transposable elements in Southern autoradiographs is proposed. PMID- 2560697 TI - [CT-guided fine needle aspiration biopsy of abdominal masses]. AB - Fine needle aspiration biopsy were made under CT guidance of 50 cases of intra abdominal masses, including liver in 37 cases, kidney in 6, pancreas and spleen in 1 respectively, and 5 biopsies were made of tumors of the abdominal cavity. The overall diagnostic accuracy was 92%, and the accuracy for malignancies was 84.6%. No complications occurred related to the procedure. The relevant factors for precise biopsy safety and the therapeutic value of this technique were discussed. It was concluded that CT-guided fine needle aspiration biopsy is precise in localization, it is a safe approach yielding satisfactory diagnostic accuracy. PMID- 2560699 TI - [Radiological study of lumbosacral nerve root sheath cyst (analysis of 65 cases)]. AB - 65 cases of nerve root sheath cyst of lumbosacral region were found out of 544 cases of myelography, 43 were confirmed by operation. The main radiologic features were: round or oval in shape in 28 cases; cylindrical or fusiform in 14; and mixed form in 13. In 10 cases, the contrast medium was seen to extend in an irregular pattern along nerve root sheath. PMID- 2560698 TI - [Dynamic CT scan of hepatic tumors]. AB - Dynamic CT manifestations in 44 cases of hepatic tumors proved by pathology were reviewed. 30 were hepatoma, 6 metastatic tumor, 7 cavernous hemangioma and 1 adenoma. 50 ml of contrast medium were administered as a bolus injection, and scanning was performed at 15, 25, 60, 120 and 300 sec after injection. The tumor enhancement patterns were divided into 3 types based on time-density curve. Type 1 manifested rapid and prolonged marked tumor enhancement; Type 2 was characterized by a rapid rise followed by a rapid decline in tumor density; In type 3 there was no significant change over time in density of the tumor. All cavernous hemangiomas belonged to type 1. Type 2 and 3 were not specific. Parafocal enhancement which occurred in 83.3% of primary hepatic carcinoma in this article and without false positive cases was considered by the authors to be pathognomonic of hepatoma. Evaluation was also made of dynamic CT in treatment planning of hepatic tumors. PMID- 2560700 TI - [A correlative study of sex hormonal receptor status and mammographic appearance in primary breast cancer]. AB - The relation between the sex hormonal receptor status and the mammographic appearance of the primary breast cancer in 162 patients was studied. The hormonal receptor status of breast cancer could be inferred by its shape and size as seen on the mammogram. The hormonal receptor level tended to be high tumors with prominent spiculate shadows or when the size of the tumor was small regardless of its shape, but lower in the majority of tumors which were large and lumpy with less hair-like projections or devoid of spicules. The mechanism of this phenomenon has been investigated. In addition, 119 patients with mammography taken of the opposite breast were analysed and no correlation was found between Wolfe's parenchymal pattern and hormonal receptor status. The authors suggested that mammography might be a simple and reliable method of assess the hormonal receptor status in breast cancer. PMID- 2560701 TI - Circadian rhythm parameters of endocrine functions in elderly subjects during the seventh to the ninth decade of life. AB - The circadian rhythm of 17 endocrine parameters (ACTH, aldosterone, cortisol, C peptide, DHEA-S, FSH, growth hormone, insulin, LH, 17-OH progesterone, prolactin, testosterone, total T3, total T4 and TSH and estradiol and progesterone in women only) were studied in 63 clinically apparently healthy men (124 profiles) and 86 women (154 profiles) during the 7th to 9th decade of life. The subjects lived under very uniform conditions in a home for the aged with their daily schedule standardized by institutional routine with rest at night on the average from 21:30 to 06:30 local time and 3 daily meals at 08:30, 13:00 and 18:30. Blood was drawn over a 24-h span at 4-h intervals. Circadian periodicity was ascertained and the rhythm parameters quantified by cosinor analysis. In clinically healthy elderly subjects, circadian periodicity persisted in most parameters studied well into the 9th decade of life. The timing of the circadian rhythm was comparable between subjects in their 7th decade and 9th decade of life with the exception of cortisol and DHEA-S, which showed a phase advance with advancing age. A decrease in circadian amplitude is limited during this part of the human life span to only a few of the functions investigated and with the exception of prolactin in the women, a decrease in amplitude did accompany a decrease in MESOR. PMID- 2560702 TI - [Hemodynamic study of a single oral dose of enalapril in patients with heart failure]. AB - Clinical and hemodynamic studies were carried out for the purpose of evaluating the effects of enalapril in the treatment of chronic heart failure. Enalapril 10 20 mg was given once to 10 patients with moderate to severe congestive heart failure (coronary 6, hypertensive heart disease 2 and idiopathic congestive cardiomyopathy). Hemodynamic studies were done on the first day after drug administration and the same dose was maintained for 2 weeks. The results showed a decrease of mean BP of 18.29% (P less than 0.01), CVP 40.4% mPAP 23.9%, PCWP 41.2%, SVR 39.9% and PVR 41.3% respectively (P less than 0.001). Cardiac index increased 44.16% (P less than 0.001). The therapeutic action lasted more than 24 hours after a single dose. No further drop of BP was found weeks later, 6 patients in NYHA class III and 2 in class IV improved to class II. No severe side effects were found. The results showed that enalapril is well tolerated after a single dose and its therapeutic action lasts 2 weeks with improvement both in hemodynamics and subjective symptoms. PMID- 2560704 TI - [Plasma level of cyclic nucleotides in patients with congestive heart failure and their clinical significance]. AB - Plasma concentrations of cyclic nucleotides were determined by radioimmunoassay in 196 cardiac patients. The results showed that plasma concentrations of cyclic AMP and cyclic GMP increased with the deterioration of cardiac function. Plasma cyclic GMP concentrations were correlated positively to the cardiothoracic ratios (CTR), PEP/LVET, and the intracardiac diameters measured by echocardiography respectively (r = 0.55 r = 0.50, and r = 0.55, P less than 0.001), and were correlated negatively to the left ventricular ejection fractions (LVEF) and LV short axis shortenings (LVSAS) (r = -0.53 and r = -0.50 respectively, P less than 0.001). There were only weak correlations between plasma cyclic AMP concentrations and CTR, PEP/LVET ratios, LVEF and LVSAS (r = 0.35, r = 0.38, r = 0.28, and r = -0.31, respectively, P less than 0.01). The correlations between plasma cyclic GMP concentrations and mean pulmonary artery pressures (r = 0.48, P less than 0.05) and mean left atrial pressures (r = 0.55, P less than 0.01) were also significant. In patients who had received cardioversion and valve replacement, plasma cyclic GMP concentrations decreased significantly. Thus the plasma concentrations of cyclic nucleotides could be used as a useful parameter in evaluation of cardiac function and therapeutic effect. PMID- 2560703 TI - Dietary fiber in foods: options for diabetes education. AB - Diet therapy for the treatment of diabetes often dictates that individuals limit their intake of foods high in fats or sugars. The addition of fibrous foods may be appealing to this population. During the past two decades, clinical investigations have attempted to characterize the benefits of dietary fiber for diabetes. Interpretation of the data and implications for practical application have sometimes been controversial. As results of more clinical trials appear in the literature, a convergent validity is emerging. Evidence that soluble dietary fibers are a food component associated with health benefits is supported by recent investigations. A flexible approach for inclusion of dietary fiber in the diets of individuals with diabetes may be used. PMID- 2560705 TI - [Sodium and calcium cation transport of erythrocytes in essential hypertension]. AB - The sodium and calcium transport of erythrocyte and the influencing factors were studied in essential hypertensive (EH) subjects. The result showed that plasma endogenous digitalis-like compound (EDLC) increased and sodium pump depressed in some EH patients, but there were no parallel correlation between EDLC and sodium pump. The patients with normal sodium pump mainly showed their maximal Ca2+ pump activity and decreased calmodulin (CaM) content of erythrocyte. Thus there may be different types of ion transport defect in EH, and the abnormalities of these cation transports have an important role in the pathogenesis of EH. PMID- 2560706 TI - [Hemodynamic changes caused by a single intravenous dose of amrinone in congestive heart failure]. PMID- 2560707 TI - Analysis of competence: receptors for fibroblast growth factor in early Xenopus embryos. AB - Xenopus ectodermal cells have previously been shown to respond to acidic and basic FGF by differentiating into mesodermal tissue. In the present study, ectodermal explants from Xenopus blastulae were shown to have high affinity binding sites for 125I-aFGF (Kd = 1.4 X 10(-10) M). The total number of sites, determined by Scatchard analysis, was 3 X 10(8) per explant (surface area of approximately 1 mm2). Two putative receptors of relative molecular mass 130,000 and 140,000 were identified by chemical crosslinking to 125I-aFGF. Both acidic and basic FGF, but not TGF beta 2, could compete for affinity labelling of these bands. The receptor density at the cell surface parallels the developmental competence of Xenopus animal pole cells to respond to FGF. Receptors are present at highest density in the marginal zone but are not restricted to cells in this region. PMID- 2560708 TI - Insulin-like growth factor II may play a local role in the regulation of ocular size. AB - The ultimate size and shape of the eye has a profound influence on its refraction and function. However, the role of growth factors in normal ocular development is poorly understood. Insulin-like growth factors IGF-I and -II have major effects on cell growth and differentiation in tissue culture. Recently their importance for in vivo development has been studied; IGF-II is predominant prenatally, with a probable local role in the differentiation of some mesodermally derived tissues. Ocular development and size is partially dictated by the condensation of the outer collagenous scleral coat (the 'white') of the eye from orbital mesoderm. We investigated IGF-II expression and IGF-II receptor distribution during normal ocular development in the mouse fetus using in situ hybridization and immunohistochemistry. IGF-II mRNA was expressed by the loose mesenchymal orbital tissue as it differentiated to form the sclera, but not in the compact mature sclera or cornea, or in the ectodermally derived retina or skin. IGF-II gene expression was seen in the orbit at E14, reached a peak just before parturition and then declined to background levels after birth. Similarly, type 2 IGF receptors were shown with immunohistochemistry to be present on developing scleral cells and to be modulated in parallel with IGF-II mRNA expression. We suggest the IGF-II expression by differentiating cells that compact to form the collagenous ocular coat plays a local role in determining the ultimate shape and size of the developing eye. PMID- 2560710 TI - [Primary tumors of the central nervous system from material of the Department of Pathological Anatomy--Plovdiv, 1975-1984]. AB - 8136 deceased at the clinics of the Higher Medical Institute "I. P. Pavlov" for the period of 1975-1984 are studied. 265 of the deceased (3.28%) have tumours of the central nervous system (cns), among which astroglial tumours predominate (73.21%), while nonastroglial tumours represent 26.79%. The largest is the number of astrocytomas II degree--99 cases, followed by astrocytomas III degree--47 cases, multiform glioblastomas--28 cases and astrocytomas I degree--20 cases in accordance with differentiation of astroglial tumours. A larger part of astroglial tumours are localized supratentorially (86.6%) as the mature age is manifested mainly (31-60 years)--61.34% (p less than 0.001). The male sex predominates (61.84%) over the female sex (38.16%) (p less than 0.001) among the decreased with astroglial tumours. The cerebral oedema--64.43% is the most frequent immediate cause for the death of these patients. Menigiomas (42.25%) predominate among 71 patients with nonastroglial tumours, followed by meduloblastomas and ependimomas--15.49% and respectively 12.68%, oligodendrogliomas 8.45%. The remaining histogenetic types of papillomas of choroid plexus, angioreticulomas (hemangioblastomas according to the classification the World Health Organization, 1979), craniopharingiomas are more rarely encountered tumours. PMID- 2560711 TI - [Current concepts of the transport of proteins in mitochondria]. PMID- 2560709 TI - Biochemical differentiation in a mutant of Dictyostelium discoideum defective in cyclic AMP chemotaxis and in intercellular cohesion. AB - A temperature-sensitive mutant of Dictyostelium discoideum has been isolated based on its lack of chemotaxis toward cyclic AMP at the restrictive temperature, 27 degrees C. The mutant develops normally at the permissive temperature, 22 degrees C, but fails to aggregate or complete development at the restrictive temperature. The temperature-sensitive phenotype can be bypassed by allowing cultures to grown into late log phase or to starve for 60-90 min at 22 degrees C prior to a shift to 27 degrees C. At 27 degrees C, the mutant overproduces cell surface cyclic AMP receptors of both high and low affinity and is capable of spontaneous oscillations in light scattering in cell suspensions. Despite its complete lack of morphological development, the mutant undergoes extensive biochemical differentiation. At the onset of starvation, it shows increased levels of N-acetylglucosaminidase, it express cyclic AMP receptors at the normal time and, although somewhat slowly, suppresses those receptors as if aggregation had been achieved. Metabolic pulse labellings with [35S]methionine revealed that the mutant at 27 degrees C displays the same changes in the patterns of newly synthesized proteins observed during the vegetative-to-aggregation and the aggregation-to-slug stages of normal development. The only clear difference from wild type was the failure of the culmination-stage isozyme of beta-glucosidase to appear. The mutant is defective in establishment of intercellular cohesion mechanisms, correlated with poor agglutination by concanavalin A, at the restrictive temperature. The properties of the mutant place severe constraints on models regarding the role of chemoreception and intercellular cohesion in regulation of gene expression. PMID- 2560712 TI - The relationship between cortisol and ACTH in patients with Cushing's disease following neurosurgery or pharmacotherapy. PMID- 2560714 TI - Internalization of beta-adrenergic receptor in A431 cells involves non-coated vesicles. AB - To characterize the mechanism of internalization of beta-adrenergic catecholamine receptors on human epidermoid A431 carcinoma cells, their distribution was analyzed by immunocytochemistry using the monoclonal anti-receptor antibody BRK2. In preconfluent cultures, the receptors appeared to be randomly distributed on the cell surface. Exposure to the agonist isoproterenol induced an overall decrease in the number of cell surface receptors as determined by binding experiments and visualized by immunofluorescence. When cells were incubated at 4 degrees C with BRK2 and anti-mouse IgG-gold and then transferred at 37 degrees C, non-coated invaginations and vesicles were labeled. The addition of isoproterenol resulted in an increased rate of internalization of the receptor-BRK2-anti-IgG gold complex. When incubation with the two antibody reagents was prolonged (with or without isoproterenol), non-coated vesicles fused in the endosomal compartment, and receptors were transferred to multivesicular bodies and lysosomes. At no stage in this process was there any indication that clathrin coated pits or vesicles participated. Furthermore, we found that an intracellular potassium depletion treatment known to inhibit endocytosis, did not affect the normal pattern of desensitization of beta-adrenergic receptors. PMID- 2560713 TI - Direct adhesion to type I and homotrimer collagens by breast carcinoma and embryonic epithelial cells in culture: a comparative study. AB - A continuous cell line of neoplastic cells derived from ductal infiltrating carcinoma of the human breast (8701-BC), was assayed for its ability to adhere to collagen substrates. The collagens used were regular type I and type I homotrimer isolated from primary breast carcinomas. Comparative studies were performed using an embryonic epithelial cell line derived from human intestine (Int. 407). The neoplastic cells adhere equally well to both collagens, while the embryonic epithelial cells recognized only the homotrimer. Some receptor diversity was recognized in the adhesion of the two cell lines to homotrimer collagen. The data demonstrate a functional difference between type I and homotrimer collagen with regard to cellular recognition and attachment. In addition, the data suggest that oncogenic transformation of breast epithelial cells promotes their adhesive properties to interstitial collagens and that this may be relevant to their increased potential to invade host tissue. PMID- 2560715 TI - Disorganization of mitosis in HeLa cells by deuterium oxide. AB - We assessed, by light and electron microscopy, the influence of deuterium oxide on the dynamics of mitosis and on the morphology of the mitotic apparatus in HeLa cells grown in vitro. A 2-h incubation of HeLa monolayers with low concentrations of D2O (1%-25%) in the medium increased the frequency of multipolar divisions up to 20 times the control level. Substitution of 10% and 25% D2O for H2O induced changes in the proportions of mitotic phases. These changes could be fully reversed to the control pattern after 1 h of recovery in non-deuterated medium. Fifty % D2O strongly inhibited, and 75% D2O blocked the cell cycle before prophase and at (pro-)metaphase. In cells treated with 50% D2O conspicuous morphological changes of the interphase chromatin as well as ultrastructural abnormalities of all mitotic phases were regularly observed. Overall, these results confirmed the antimitotic activity of deuterium oxide and revealed that it could also influence the cell cycle before mitosis. It is suggested that interference with diverse cellular constituents rather than a specific influence on microtubule turnover could be responsible for the disorganization of the cell cycle in HeLa cells by D2O. PMID- 2560717 TI - Inhibition of phosphotyrosine phosphatases reveals candidate substrates of the PDGF receptor kinase. AB - In normal fibroblasts stimulated by platelet derived growth factor (PDGF), PDGF receptors are transiently phosphorylated on tyrosine and represent the major phosphotyrosine containing protein. The phosphate of the phosphotyrosine groups turns over rapidly, and extensive evidence indicates a dynamic balance between phosphorylation and dephosphorylation reactions. Thus, the effect of an inhibitor of phosphatases, orthovanadate, on the pattern of the tyrosine phosphorylations induced by PDGF in Swiss 3T3 fibroblasts was investigated. Western blot analysis with antibodies against phosphotyrosine indicated that whereas in unstimulated cells no phosphotyrosine containing proteins were detected, treatment of cells with orthovanadate alone elicited the slow phosphorylation of several proteins including a 170 kDa component that was recognized to be the phosphorylated PDGF receptor. Addition of PDGF to cells shortly pretreated with vanadate highly increased the intensity of the 170 kDa band corresponding to the phosphorylated receptor and caused its stabilization during time. In addition, the phosphorylation on tyrosine of other proteins (molecular mass 116, 80, 73, 60, 50 and 39 kDa) was also induced. Both the receptor and the other tyrosine phosphorylated proteins appeared to be associated with the detergent insoluble matrix. PMID- 2560716 TI - Transformation of Golgi membrane into the envelope of herpes simplex virus in rat anterior pituitary cells. AB - Envelopment of herpes simplex virus type-1 (HSV-1) was investigated in relation to membrane differentiation in dissociated anterior pituitary cells. The number of cells stained positively with anti-HSV-1 serum was increased from 16 h to 31 h post infection. During this period, electron microscopy revealed that a number of nucleocapsids (unenveloped particles) were accumulated in the Golgi area, where they frequently became surrounded by a double membrane of short Golgi cisternae or by one with a Golgi associated endoplasmic reticulum lysosome (GERL)-like structure. The inner membrane of the cisterna surrounding the nucleocapsids showed regional specialization which was characterized by increased thickness and electron opacity. Acid phosphatase activity, a marker for GERL or trans Golgi cisternae, appeared in the cytoplasmic short cisternae surrounding the nucleocapsids, whereas glucose-6-phosphatase activity, a marker for the nuclear envelope or for endoplasmic reticulum, was not demonstrated in such cisternae. Monoclonal antibody against glycoprotein gD revealed that gD was localized in the trans Golgi membrane as well as in the envelope of the virion. The antibody binding sites were highly concentrated in the area where Golgi membranes showed increased opacity. Furthermore, nucleocapsids were surrounded exclusively by gD positive cisternal (Golgi or Golgi-derived) membranes. Thus, our results indicate that the envelope of HSV is derived from trans Golgi cisterna (GERL), and that some viral components, including gD, destined for the envelope may be assembled initially in the Golgi membrane, which is thereby transformed into the envelope of the virus. PMID- 2560719 TI - Glioblastoma multiform in a dermoid cyst of the ovary. AB - Malignant transformation is reported in less than 2% of benign cystic teratomas. Although all the elements can undergo this transformation, it is most often seen in squamous epithelium. The malignancy of neural elements is probably the least common event, with only one case previously reported. A case of glioblastoma multiform in a benign cystic teratoma is presented. Its neural derivation was supported by an immunohistochemical staining specific for Glial Fibrillary Acidic Protein (GFAP). Despite very conservative surgery without adjuvant therapy, the patient remains alive and symptom free more than 3 years later. PMID- 2560718 TI - Pathologic complete response of advanced ovarian mixed mesodermal tumor to cisplatin, adriamycin and dacarbazine: a case report. AB - Presented is a case report of a stage IIIC primary ovarian mixed mesodermal tumor with gross residual disease following aggressive cytoreductive surgery. Following twelve courses of Cisplatin, Adriamycin and Dacarbazine, second-look laparotomy confirmed a pathologic complete response. The patient is now free of disease at 17+ months. PMID- 2560720 TI - Metastatic cancer spread at the laparoscopic puncture site. Report of a case in a patient with carcinoma of the ovary. Case report. AB - The aim of this study is to draw attention to the possibility of the occurrence of a metastatic spread at the laparoscopic entrance site in patients suffering from a carcinoma of the ovary. This event has rarely been mentioned in medical literature before. A case is presented here in which a localized tumoral lesion appeared after a laparoscopic staging study in the abdominal wall, exactly at the site used for the introduction of the accessory laparoscopic trocar in a patient suffering from a carcinoma of the ovary. PMID- 2560721 TI - The changing face of "myeloma". PMID- 2560722 TI - [The electrical activity of the hippocampus in rats centrally administered GABA agonists and antagonists]. AB - In unrestrained rats, frequency ranges of dorsal hippocampus electrograms were studied after intraventricular injections of the GABA, muscimol, baclofen, bicuculline and picrotoxin. The data obtained corroborates the idea of an interrelationship between the agonists and antagonists of the GABA receptors. A possibility of participation of these receptors in the formation and/or modulation of total electrical hippocampal activity, is discussed. PMID- 2560723 TI - [Signal transduction system of hormones and cellular response, with special reference to vasopressin and ACTH]. PMID- 2560725 TI - [The structure and function of endothelin]. PMID- 2560724 TI - Recent progress in neuroimmunology research. PMID- 2560726 TI - Beta-adrenoceptors desensitization may modulate catecholamine induced insulin resistance in human pheochromocytoma. AB - Catecholamines acutely exert a pronounced insulin-antagonistic effect, which is mediated by beta-adrenergic receptors stimulation. Nevertheless, several patients with pheochromocytoma fail to exhibit an overt diabetic syndrome, in spite of steadily elevated plasma levels of catecholamines. This prompted us to investigate a 16 years old male patient, bearing an extra-adrenal pheochromocytoma, who displayed a slightly impaired glucose tolerance to oral glucose tolerance test, whereas fasting and post-prandial blood glucose, as well as glycaemic response to intravenous glucagon, were in the normal range. Peripheral insulin sensitivity, as evaluated by intravenous insulin tolerance test, was slightly decreased. Supine norepinephrine plasma levels were steadily upon 9 ng/ml; plasma insulin, both fasting and post-prandial, was within the normal range. beta-adrenergic receptors density of peripheral mononuclear cells was strongly reduced when compared to controls (0.97 +/- 0.08 vs 2.82 +/- 0.37 fmol/10(6) cells), without any concomitant change of affinity. Insulin binding to circulating monocytes was reduced as well (2.38 +/- 0.27 vs 5.1 +/- 0.4%/10(7) monocytes); insulin receptor affinity was quite normal (1.7 ng/ml) and total receptor number was 9,200 sites/cell. In desensitization experiments, 1 microM isoproterenol caused only a 20% decrease of beta-adrenergic receptors density in the patient's cells (70% decrease in controls). Six months after surgery, all the above modifications of receptor binding, as well as the mild glucose intolerance, were almost completely reversed. Thus, high levels of norepinephrine were able to induce a decrease of both beta-adrenoceptor and insulin receptor binding, together with a marked reduction of in vitro agonist-induced redistribution of beta-adrenergic receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2560727 TI - GTP and its non-hydrolysable analogues stimulate polyphosphoinositide hydrolysis in plasma membranes of rat hepatocytes. PMID- 2560728 TI - Effect of glucose analogues on yeast adenylate cyclase in vitro. PMID- 2560729 TI - Cancer reversal: regulation of phosphotyrosine phosphatase. PMID- 2560730 TI - Protein kinases in human lymphocytes: electrophoretic study of multiple forms. PMID- 2560731 TI - 2-Chloroadenosine-sensitive calcium channels in synaptosomes: effects of Ba2+. PMID- 2560732 TI - On the molecular basis of memory. PMID- 2560733 TI - A new method of screening receptor cDNAs: influence of plasmid competition on receptor expression. PMID- 2560734 TI - Effects of vanadate on receptor-mediated endocytosis of asialoglycoproteins in isolated rat hepatocytes. PMID- 2560735 TI - Effect of electromagnetic induction on impulse conduction in the frog nerve muscle preparation. PMID- 2560736 TI - Cyclic AMP-dependent protein phosphorylation in primate kidney. PMID- 2560737 TI - Oxidants and vascular smooth muscle. PMID- 2560738 TI - Brain microsomes bind ryanodine and contain ryanodine-sensitive calcium channels. PMID- 2560739 TI - Interaction of MnATP and peptide substrate with insulin receptor tyrosine kinase. PMID- 2560740 TI - Regulation of insulin receptor tyrosine kinase by metabolic intermediates. PMID- 2560741 TI - Effect of Li+ on the secretion of HCO3- in rat fundic tissue. PMID- 2560742 TI - Investigations on the cytoprotective mechanisms of thiocyanate in rat gastric mucosa. PMID- 2560743 TI - Hepatic phosphatidate phosphohydrolase activity in acute and chronic alcohol-fed rats. PMID- 2560744 TI - Active sites of yeast H(+)-ATPase studied by directed mutagenesis. PMID- 2560745 TI - Upstream regulation of Saccharomyces cerevisiae adenylate cyclase. PMID- 2560746 TI - Do cells contain discrete pools of inositol lipids that are coupled to receptor activation? PMID- 2560747 TI - Receptor-operated calcium channels in human platelets. PMID- 2560748 TI - Gap junctions and intercellular communication: topology of the major junctional protein of rat liver. PMID- 2560749 TI - Activation of phosphatidylcholine-specific phospholipase C in cell growth and oncogene transformation. PMID- 2560750 TI - Localized surface activity of torso, a receptor tyrosine kinase, specifies terminal body pattern in Drosophila. AB - The subdivision of the Drosophila body into distinct terminal and central domains depends on the torso (tor) protein, a putative receptor tyrosine kinase that is active at both ends of the early embryo. We show that the tor protein is uniformly expressed along the surface membrane of early embryos despite its localized activity at both poles. Further, we present evidence that polarized activity of this protein depends on other terminal gene functions, one of which may be a localized extracellular ligand generated during oogenesis. Finally, using the temperature-sensitive gain-of-function mutation torRL3, we show that different levels of active tor protein can specify distinct portions of the terminal pattern. Thus, we argue (1) that for functions as a ubiquitous surface receptor that is activated by a spatially restricted ligand, and (2) that localized activity of the tor kinase may generate one or more gradients of intracellular signals that control body pattern. PMID- 2560751 TI - Identification of MRF4: a new member of the muscle regulatory factor gene family. AB - We have identified a rat cDNA encoding MRF4, a new member of the muscle regulatory factor gene family that includes MyoD1, myogenin, and Myf-5. MRF4 encodes a predicted 27-kD protein that contains a conserved helix-loop-helix motif, which is a common feature of this gene family. Northern analyses indicate that MRF4 is expressed solely in skeletal muscle tissue but is not detected in most embryonic muscle cell lines. Transfection of MRF4 into C3H10T1/2 fibroblasts produces stable myogenic lineages at frequencies that are equal to or greater than those obtained when MyoD1 or myogenin are introduced into these cells. Expression of the MRF4 cDNA leads to expression of the endogenous MyoD1 and myogenin genes, although C3H10T1/2 cells expressing MyoD1 or myogenin cDNAs do not express MRF4. Interestingly, the endogenous MyoD1 and myogenin genes are negatively regulated by serum and by purified growth factors since MRF4 transfected C3H10T1/2 cells activate MyoD1 and myogenin expression only in mitogen-depleted, differentiation-induced muscle cultures. The myofiber-specific expression pattern of MyoD1 and myogenin in these cells suggests that the primary role for this muscle regulatory factor gene family may be in regulating specific terminal differentiation events that are crucial for normal skeletal muscle development. PMID- 2560752 TI - The mutated, myeloid cell-specific growth factor receptor v-fms transforms avian erythroid but not myeloid cells. AB - In avian hematopoietic cells, transformation by tyrosine kinase oncogenes is restricted to the erythroid lineage. To study the mechanism of this striking target cell specificity, we constructed an avian retrovirus correctly expressing the feline v-fms protein, an oncogenic version of the myeloid-specific CSF-1 receptor. Similar to other tyrosine kinase oncogenes (erbB, src, fps, sea), the v fms oncogene induced progenitor cells to self-renew in an erythropoietin independent manner. Spontaneous differentiation of the transformed cells was arrested by v-erbA. v-fms failed to induce transformation of myeloid cells but caused myeloid cells transformed by the v-myb oncogene, as well as normal macrophages, to proliferate independent of cMGF (chicken myelomonocytic growth factor). Unlike the other tyrosine kinases, v-fms did not induce cMGF secretion in myb-myeloblasts, suggesting a nonautocrine mechanism of growth factor independence. PMID- 2560753 TI - Precise excision of telomere-bearing transposons during Oxytricha fallax macronuclear development. AB - In ciliated protozoa, development of the macronucleus from a copy of the germ line micronucleus involves elimination of a large number of sequences by DNA splicing akin to precise excision of transposons. The known examples of such internal eliminated sequences (IESs) are not repetitive. The telomere-bearing elements (TBE1s) of Oxytricha fallax are a family of transposons. We show that two particular TBE1s are also IESs. TBE1-1 and TBE1-2 disrupt a micronuclear region that codes for macronuclear DNA. A variety of tests indicates that each TBE1 and one copy of the flanking target repeat is absent from most, if not all, molecules of the macronuclear DNA, as if the TBE1s were precisely excised during macronuclear development. Three alternative explanations for the absence of TBE1 1 and TBE1-2 from the macronuclear DNA were tested. First, because two other highly homologous versions of that DNA are also found in the macronucleus, recombination between versions during or after macronuclear development could have bypassed the elements. Recombination in the regions flanking the elements was not detected. Second, micronuclear DNA blots show no evidence of a micronuclear counterpart of the macronuclear region that lacks TBE1-1. Third, TBE1-2 was demonstrated in two sexually independent cell lines. This shows that it pre-existed in the germ line, as opposed to having transposed into the micronuclear DNA subsequent to the generation of the macronucleus of the vegetative line that is usually studied. We conclude that TBE1-1 and TBE1-2, and possibly many of the other approximately 1900 micronucleus-limited TBE1s are excised as IESs during macronuclear development. These transposons appear to enjoy the luxury of relaxed constraints on family expansion, because they are removed from the genome before it is expressed. We discuss the possibility that all IESs are transposon-derived, that all are excised by transposition machinery, and that linear excision products are early intermediates in transposition. PMID- 2560754 TI - In vitro reconstitution of functional yeast U2 snRNPs. AB - A system for the functional reconstitution of yeast U2 snRNPs using synthetic U2 RNAs is described. We use oligonucleotide-directed RNase H cleavage to specifically deplete yeast extracts of their endogenous full-length U2 snRNA and consequently inactivate pre-mRNA splicing activity. The subsequent addition of synthetic yeast U2 RNAs, derived by in vitro transcription (T7U2 RNAs), to these oligonucleotide-treated extracts efficiently reconstitutes their ability to splice pre-mRNA. The use of deletion derivatives of the T7U2 RNA has demonstrated that the region downstream from the conserved Sm-binding site sequence in the yeast U2 RNA is not absolutely required for pre-mRNA splicing activity in vitro. Furthermore, we found that both human and rat U2 RNAs can function in yeast extracts. We also show that point mutations in the yeast U2 RNA can be analyzed using the in vitro reconstitution system. Allele-specific suppression of mutations in pre-mRNA branch site sequence is observed when the appropriate compensatory mutations in the branch site recognition region of the T7U2 RNA are introduced. Finally, we present a model for the interaction of the U2 and U6 snRNAs during pre-mRNA splicing. PMID- 2560755 TI - In vitro assembly of yeast U6 snRNP: a functional assay. AB - U6 small nuclear RNA (snRNA) is the most highly conserved spliceosomal RNA, and it has been postulated to have a fundamental role in pre-mRNA splicing. To elucidate this role, we developed an in vitro system for reconstituting the functional U6 small ribonucleoprotein (snRNP). Treating splicing extracts with an oligonucleotide complementary to the central domain of U6 snRNA leads to both RNase H cleavage of the endogenous U6 snRNA and loss of splicing activity. Yeast U6 RNA, synthesized in vitro using T7 RNA polymerase, is then added to the oligonucleotide-treated extract, and restoration of splicing activity is monitored by the subsequent addition of substrate pre-mRNA. Addition of full length, unmodified T7U6 snRNA (113 nucleotides) to oligonucleotide-treated extracts restores splicing activity efficiently. Using U6 RNA transcripts truncated at their 3' ends, we show that large deletions (39 nucleotides) produce molecules that are unable to restore splicing activity in vitro and cannot interact with the endogenous U4 snRNA or form a mature spliceosome. Finally, we show that substitution of the invariant G81 with C within the T7U6 RNA abolishes its ability of restoring splicing activity. Although the U4/U6 snRNP forms correctly, mature spliceosomes do not assemble. PMID- 2560756 TI - A general method for the induction and screening of antisera for cDNA-encoded polypeptides: antibodies specific for a coronavirus putative polymerase-encoding gene. AB - A prokaryotic vector, pGE374, containing the recA and lacZ genes, out-of-frame, was used for the expression of cDNA derived from the putative polymerase-encoding gene of the coronavirus mouse hepatitis virus strain A59 (MHV-A59). The pGE374/viral recombinant vector generates a tripartite bacterial/viral protein composed of a segment of the RecA protein at the N terminus, the coronaviral sequences in the middle, and an enzymatically active beta-galactosidase at the C terminus. Rabbits immunized with such recombinant proteins generated antibodies to the MHV-A59 portion of the tripartite protein. Because the MHV-A59 polymerase proteins have been difficult to identify during infection, we used a novel method to demonstrate the viral specificity of the antiserum. The viral cDNA was excised from the expression vector, and transferred to a pGem vector, downstream from and in-frame with a portion of the cat gene. This construct contained a bacteriophage RNA polymerase promoter that enabled the cell-free synthesis of a fusion protein that was used to verify that antibodies were generated to the expressed viral DNA. This strategy was shown to successfully result in the specific generation of antibodies to the encoded information of the viral cDNA. Furthermore, this method has general applicability in the generation and characterization of antibodies directed against proteins encoded in cDNAs. PMID- 2560757 TI - Graduated resistance to G418 leads to differential selection of cultured mammalian cells expressing the neo gene. AB - The effects of Geneticin (G418) selection on the growth and survival of cultured mammalian cells expressing the neomycin-resistance gene (neo) were studied by the analysis of cell clones from two retroviral neo vector-infected populations. We found a correlation between the neo expression level and growth rates in medium containing varying G418 concentrations. This relationship permits the use of differential selection schemes for the isolation of rare cells with increased expression. Comparison, by clone sampling, of vector-positive populations before and after selection with a G418 concentration in the range usually used for selection, showed different expression level and vector copy number distributions for the population infected with the vector of lower LTR activity, but not for the other. Such biasing effects of G418 selection may be important when selected cells are used for quantitative studies of gene expression. PMID- 2560758 TI - Structural alignment of retroviral protease sequences. AB - All the known retroviral proteases can be accommodated into the structural alignment of the Rous sarcoma virus and human immunodeficiency virus proteases. PMID- 2560759 TI - [Use of humans and animals of agricultural products grown on soils fertilized with wastes from swine breeding complexes]. PMID- 2560760 TI - [Therapeutic possibilities in the climacteric]. PMID- 2560761 TI - [Neurophysiologic processes in the production of climacteric hot flushes]. PMID- 2560762 TI - [The modification of thyroid dysfunction by hormone substitution in the climacteric]. PMID- 2560763 TI - Secondary lymphomas: a review on lymphoproliferative diseases arising in immunocompromised hosts: prevalence, clinical features and pathogenetic mechanisms. AB - This paper reviews the major information on lymphoproliferative diseases developing in primary and acquired immunodeficiencies, in organ allograft recipients, and in different diseases with immune impairment such as rheumatoid arthritis, angioimmunoblastic lymphadenopathy, and Hodgkin's disease (secondary lymphomas). The hypothetical role of Epstein-Barr virus (EBV) in the pathogenesis of lymphoproliferative diseases in immunocompromised hosts has come from the examination of lymphoma cells or tissues for Epstein-Barr nuclear antigen (EBNA), or carriage of the viral genome, and will be extensively reviewed. The characteristics and the prognosis of high-grade lymphomas developing in the acquired immunodeficiency disease (AIDS) will be analyzed, together with their pathogenetic mechanisms, with particular emphasis on the constant presence in the lymphoma cells (mostly of Burkitt-type) of the c-myc oncogene rearrangement and activation. The principal methods of study of secondary lymphomas and major attempts at therapy will reviewed as well. PMID- 2560764 TI - Activation of mouse T lymphocytes by a monoclonal antibody to a developmentally regulated surface aminopeptidase (THAM). PMID- 2560765 TI - Influence of detergents and pH on the isolation, purification and molecular properties of alpha-galactosidase-C2 from germinating guar (Cyamopsis tetragonolobus). AB - alpha-Galactosidase was isolated from germinating guar. The extract also contained small amounts of alpha-mannosidase and beta-mannosidase activities. The fractionation of the enzyme extract with ammonium sulphate (75% saturation) resulted in the appearance of all the three enzymes in a floating lipid complex. The inclusion of detergents such as Triton X-100 and sodium deoxycholate in the extraction medium failed to prevent the appearance of these enzymes in the floating lipid complex. However, by using acetone powder of the seedlings, alpha galactosidase could be sedimented with ammonium sulphate. The presence of detergents in the extraction medium affected the molecular properties of the enzyme. Using a set of carefully selected conditions alpha-galactosidase was purified to apparent homogeneity. Analytical ultracentrifugation and gel filtration studies of the purified enzyme showed association-dissociation phenomenon as a function of pH and temperature. The effect of pH on the association-dissociation indicates the predominance of electrostatic interactions in the association of subunits. PMID- 2560766 TI - Lipoprotein profile during perchlorate toxicity. AB - Perchlorate administration to rats for 45 days alters the lipoprotein profile in plasma. The levels of cholesterol, phospholipids and triglycerides in HDL, LDL and VLDL fractions are significantly increased in perchlorate-treated rats. Post heparin lipolytic activity of plasma of sodium perchlorate-treated rats is decreased. The risk factor, i.e. the total cholesterol/HDL cholesterol, increases in the experimental animals, indicating that the treatment of rats with perchlorate may develop the susceptibility of the animals to cardiac heart disease. PMID- 2560767 TI - The denatured states of ribonuclease-A: mechanism of denaturation by lithium chloride. AB - Denaturation of ribonuclease-A by lithium chloride has been studied using difference spectral, circular dichroic and viscometric measurements. The difference spectral results were interpreted in the light of our observations that the solvent effect of the denaturant on the tyrosyl residue is non-linear. It has been observed that (1) the lithium chloride-denatured protein contains 3% alpha-helix and 18% beta-structure, and (2) only two of the three buried tyrosyl residues are normalized in the denatured protein. PMID- 2560768 TI - Photobleaching and cyclic GMP dependences of rhodopsin phosphorylation in rod outer segment. AB - The experimental data on the cGMP decrease under continuous illumination of rod outer segment have been theoretically analysed to study the bleaching and hence the cGMP dependence of the rhodopsin phosphorylation. From the agreement of the theoretical results with the experimental observations it has been found that the rate of phosphorylation depends on the rate of cGMP hydrolysis. If the rate of cGMP hydrolysis increases the rate of phosphorylation also increases. The results of the theoretical treatment predict that (i) the presence of cGMP in rod outer segment inhibits the rhodopsin phosphorylation and (ii) rhodopsin phosphorylation process is much faster than what has been reported in the literature. PMID- 2560769 TI - [Proteiform syndrome (Proteus syndrome). Skin manifestations in a recently observed case]. AB - The case of a 4-year-old female affected by partial gigantism of the feet, syndactyly and polydactyly, partial right hemihypertrophy of buttock and lower limb, warty hyperpigmented nevus and vulvar lipoma is described. The Authors discuss about the Proteus syndrome, pointing out its rarity, the polymorphism and the problems of differential diagnosis with the Klippel-Trenaunay-Weber syndrome and with other congenital hamartomatous disorders. PMID- 2560770 TI - Coxsackie virus A 24 variant as the etiological agent of the acute haemorrhagic conjunctivitis epidemic at Vellore, in 1986. AB - Coxsackie virus A 24 variant (CA24v) was found to be the etiological agent of an epidemic of acute haemorrhagic conjunctivitis (AHC) in Vellore during the months of October and November in 1986. CA24v was isolated in 21 of 25 acutely ill patients. In addition, seroconversion was seen in 11 of the 16 patients from whom paired sera were collected. PMID- 2560771 TI - Resolution of small cell lung cancer intracranial metastases with standard dose chemotherapy. AB - Brain metastases are a well recognized problem in patients with small cell lung cancer (SCLC). Most patients are treated with whole brain cranial irradiation therapy which frequently improves symptoms. In this report we present data on two patients who achieved radiologically confirmed complete resolution of brain metastases treated with standard dose chemotherapy for SCLC. PMID- 2560772 TI - Latex examination and surgical gloves. PMID- 2560773 TI - A variant Schmidt-Ruppin strain of Rous sarcoma virus with increased affinity for mammalian cells. AB - SR-RSV-D(H), a variant virus with extremely high tropism for mammalian cells, was isolated by passage of the Schmidt-Ruppin strain of Rous sarcoma virus of subgroup D (SR-RSV-D) through hamster cells. This variant virus has acquired an altered envelope glycoprotein, encoded by the env gene, that has high affinity for receptors on the surface of mammalian cells. The variant virus transforms rat cells at about 100 times the efficiency of the parental virus, SR-RSV-D(S), as assayed by focus formation. Addition of amphotericin B (Fungizone) to the medium at a concentration of 0.2 micrograms/ml completely inhibited rat cell transformation by SR-RSV-D(H), possibly by blocking virus penetration into the cells, whereas the drug showed no inhibitory effect on transformation of chick embryo fibroblast (CEF) cells by the variant virus or on transformation of rat cells by the parental virus. The efficiency of transformation of rat cells by the variant virus was much less than its efficiency of transformation of CEF cells. Analysis of infection of rat cells suggested that the virus can infect rat cells as efficiently as CEF cells but that rat cells were not transformed by the virus as fully as CEF cells because of inefficiency of some post-penetrational step involved in viral gene expression. The finding that E1AY cells, rat cells expressing adenovirus E1A gene, were transformed by SR-RSV-D(H) as efficiently as CEF cells supports this conclusion and suggests that expression of the E1A gene in rat cells may overcome the defect in the transforming step(s) in rat cells. PMID- 2560774 TI - Augmentation of anti-tumor immunity in low-responder mice by various biological response modifiers: analysis of effector mechanism. AB - In order to elucidate the role of biological response modifiers (BRMs) in anti tumor immunotherapy, we examined their effect on the induction of anti-tumor immunity in low-responder mice which hardly exhibit anti-tumor resistance against syngeneic Rous sarcoma virus (RSV)-induced tumors, such as B10 or B10.BR mice. The anti-tumor immunity induction in the low-responder mice was 0% on immunization with mitomycin C-treated syngeneic tumor cells alone. However, if BRMs were used as an adjuvant, BCG cell wall skeleton, OK-432 or lentinan augmented the induction of anti-tumor immunity to 50%, 33% and 33%, respectively. In the low-responder mice treated with BRMs, the anti-tumor immune cells had antigen-specificity at the induction phase of in vitro restimulation but not at the effector phase of target cell lysis by the stimulated cells. When T cells were depleted from immune spleen cells just before in vitro stimulation, cytotoxicity was not induced. Furthermore, cytotoxicity was not induced if accessory cells were removed from immune spleen cells at the induction phase. However, cytotoxicity at the effector phase was not mediated by T-lymphocytes, but by non-T cells. These results suggested that the induced cytotoxicity in low responder mice was associated with the delayed-typed hypersensitivity-like effector mechanism. PMID- 2560775 TI - Prognostic significance of myeloperoxidase positivity of blast cells in acute myeloblastic leukemia without maturation (FAB: M1): an ECOG study. AB - The relationship between cell features by light microscopy and therapeutic outcome of 72 patients with acute myeloblastic leukemia (FAB M1) were investigated. The patients were divided into two groups according to myeloperoxidase-(MPO) positive percentage of blast cells, namely a low (less than 50%) and high (greater than 50%) MPO group. No remarkable morphological difference of blast cells between these two groups was observed. The 38 patients with low MPO showed a significantly lower complete response rate (CR) (52.6%) than the 34 patients with high MPO (85.3%) (p = 0.003) that included 10 CAE positive patients and 2 ANAE-positive CAE-negative patients. The low MPO group included 5 CAE-positive patients with granulocyte dysplasia and 7 additional patients with alpha naphthyl acetate esterase (ANAE) positivity who were chloroacetate esterase (CAE) negative. Low positivity of blast cells may be due to a defective enzyme expression in the former but also may be a reflection of 'monocytic' aberrant expression in the latter. The low MPO group in M1 with this heterogeneous population is less likely to achieve CR with chemotherapy, while the high MPO group with a higher CR rate suggests a more pure entity within the myeloblastic leukemias referred to as M1. After further refinement by eliminating 24 cases that were 'esterase positive' (ANAE and/or CAE), the results were still the same, suggesting that the more immature blast populations were in the low MPO group. PMID- 2560776 TI - Peripheral-type benzodiazepine receptors are involved in the regulation of cholesterol side chain cleavage in adrenocortical mitochondria. AB - In an attempt to elucidate the physiological relevance of the peripheral type of benzodiazepine receptor in adrenocortical mitochondria, we examined the effect of three different benzodiazepines (diazepam, Ro5-4864, and chlordiazepoxide) on the conversion of cholesterol to pregnenolone, the rate-limiting step in steroidogenesis, by using cholesterol-loaded mitochondria from bovine adrenal zona fasciculata. These benzodiazepines, except chlordiazepoxide, caused a dose dependent stimulation of the cholesterol side chain cleavage in the mitochondria. The stimulatory effect of Ro5-4864 was approximately 10 times more potent than that of diazepam. No inhibitory effect of YM-684 (Ro15-1788), a potent antagonist to central-type benzodiazepine receptors, was observed in the stimulation induced by diazepam and Ro5-4864. Both external calcium ion and voltage-dependent calcium channel blocker, (+)-PN200-110, were without effect on the diazepam-induced steroidogenesis. By contrast, pretreatment of mitochondria with digitonin abolished the stimulatory effect of diazepam on the mitochondrial steroidogenesis. The present results indicate that the peripheral-type benzodiazepine receptor of adrenocortical mitochondria plays an essential role in regulating cholesterol side chain cleavage without any change of calcium channels. PMID- 2560777 TI - The neurite-initiating effect of a tripeptide aldehyde protease inhibitor on PC12h cells. AB - We report here the possible involvement of a new protease in neurite initiation by PC12h cells. Addition of a leupeptin analogue (Ac-Leu-Leu-Nle-al, ALLNal) to PC12h cells on culture plates coated with collagen type I caused de novo neurite outgrowth. Other protease inhibitors (Ac-Leu-Leu-Met-al, leupeptin, E64c, E64d, soybean trypsin inhibitor, hirudin, aprotinin, diisofluorophosphate, 6 aminocapric acid, and pepstatin A) could not mimic this neurite-initiating action. ALLNal induced the initiation of one or two long neurites from the cell body, and increased the cellular level of acetylcholinesterase to an extent similar to nerve growth factor (NGF). However, ALLNal-induced neuritogenesis is different from that induced by NGF, in which many neurites are induced from a single cell body. In addition, in contrast to neurons induced by NGF, which survive for a long time, ALLNal-induced differentiation was transient, and after 48 h percentage of cells bearing neurites started to decrease. After about 120 h exposure to ALLNal, neurites had mostly disappeared and the acetylcholinesterase activity level was not as great as that produced by NGF. These results provide evidence that ALLNal and NGF elicit neurite initiation by different mechanisms, and suggest the existence of a regulatory system of neuronal differentiation through specific protease-protease inhibitor interaction. PMID- 2560778 TI - Monoclonal antibody HK4001 completely inhibits K(+)-dependent ATP hydrolysis and H+ transport of hog gastric H+,K(+)-ATPase. AB - A monoclonal antibody (designated as HK4001) was prepared against hog gastric H+,K(+)-ATPase. It dose-dependently inhibited the H+,K(+)-ATPase activity, formation of the K(+)-sensitive phosphoenzyme, and proton uptake into gastric vesicles. The H+,K(+)-ATPase activity was completely inhibited by addition of the antibody at a molar ratio of 1:2 (antibody/catalytic subunit) at pH 7.8. The maximal inhibition decreased with decrease in pH of the medium (7.8 greater than 7.4 greater than 6.2). The Fab fragment obtained by digestion of the antibody with papain was also inhibitory. The antibody did not inhibit the K(+)-dependent p-nitrophenylphosphatase or the labeling of the enzyme with fluorescein isothiocyanate. It inhibited gastric H+,K(+)-ATPase from rabbits and rats, but did not cross-react with related cation-transport ATPases (Na+,K(+)-ATPase or Ca2(+)-ATPase) or H(+)-ATPase in the multivesicular body. From these and related findings, the antibody was suggested to recognize a highly specific site on the cytosolic surface of H+,K(+)-ATPase. The conformation of the epitope was conserved after treatment with Triton X-100, but not sodium dodecyl sulfate. In addition, judging from the stoichiometry of inactivation of H+,K(+)-ATPase by this antibody, the functional unit of H+,K(+)-ATPase was suggested to be a dimer or a tetramer (not a trimer) of the catalytic unit. PMID- 2560779 TI - Effects of temperature and pH on hemoglobin release from hydrostatic pressure treated erythrocytes. AB - The release of hemoglobin from human erythrocytes hemolyzed beforehand by hydrostatic pressure, osmotic pressure, and freeze-thaw methods was examined as a function of temperature (0-45 degrees C) and pH (5.5-8.8) at atmospheric pressure. Only in the case of high pressure (2,000 bar) did the release of hemoglobin increase significantly with decreasing temperature and pH. Maleimide spin label studies showed that the temperature and pH dependences of hemoglobin release were qualitatively explicable in terms of those of the conformational changes of membrane proteins. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of membrane proteins showed the diminution of band intensities corresponding to spectrin, ankyrin, and actin in the erythrocytes hemolyzed by high pressure. Cross-linking of cytoskeletal proteins by diamide stabilized the membrane structure against high pressure and suppressed hemoglobin release. These results indicate that the disruption of cytoskeletal apparatus by high pressure makes the membrane more leaky. PMID- 2560780 TI - The importance of parathyroid hormone in inhibition of collagen synthesis and mitogenesis of osteoblastic cell. AB - Loss of bone substance is a common manifestation of hyperparathyroidism. This suggests that parathyroid hormone (PTH) plays an important role as to bone mass. To investigate the mechanism underlying this change in bone mass, I studied the effects of PTH on collagen synthesis and mitogenesis of UMR-106 rat osteoblastic osteosarcoma cells. PTH inhibits the mitogenesis of UMR-106 rat osteosarcoma cells, the half-maximal concentration being 10(-8) to 10(-7) M, which is similar to the EC50 for cyclic AMP accumulation. Cyclic AMP, whose intracellular concentration was increased by PTH, plays a role in the modulation of mitogenesis, as shown by the comparable inhibitory effects of 8-bromoadenosine 3',5'-cyclic AMP (10(-4) M), forskolin (10(-7) M), and the phosphodiesterase inhibitor, IBMX (10(-5) M). PTH, in a similar concentration range, directly inhibited collagen synthesis. Concurrent with the suppression of collagen synthesis, the amounts of a1(I) and a2(I) collagen mRNA decreased proportionately. The results show that PTH modulates collagen synthesis at the transcriptional level. I concluded that parathyroid hormone inhibits the mitogenesis of osteoblasts as well as collagen synthesis by these cells. The decreases in the number of osteoblasts and the amount of collagen synthesis contribute to the loss of bone substance in hyperparathyroidism. PMID- 2560781 TI - Relationship between numbers of alpha 2- and beta 2-adrenoceptors in teat tissue and blood cells and milkability of primiparous cows. AB - Milking characteristics were measured on 14 cows at morning milkings. After collection of blood, cows were slaughtered and teats were removed. beta 2 Adrenoceptors on teat tissue and lymphocyte membranes were identified by binding of [3H]dihydroalprenolol. For the determination of alpha 2-adrenoceptors on teat tissue and blood platelet membranes, [3H]rauwolscine was used. In teat tissue and blood cells, except for the amount of milk collected during the 3rd min, all other milking parameters were highly correlated with the ratio of beta 2 adrenoceptors to alpha 2-adrenoceptors. Good milkability appears to be associated with low ratio of beta 2-adrenoceptors to alpha 2-adrenoceptors densities in teat tissue, and a high ratio of beta 2- to alpha 2-adrenoceptors densities in blood cells. We postulated earlier that a low ratio of beta 2- to alpha 2-adrenoceptors in teat tissue of fast milking cows probably reflects changes of mainly prejunctional adrenoceptors. From our results on blood cells, we are now hypothesizing that in teat tissue prejunctional and extra junctional alpha 2- and beta 2-adrenoceptors might have been identified together, whereas alpha 2- and beta 2-adrenoceptors identified on blood platelets and lymphocytes, respectively, are reflecting by preference extra junctional adrenoceptors are activated primarily by circulating epinephrine. The high ratio of beta 2- to alpha 2 adrenoceptors observed in blood cells of fast milking cows might indicate a decreased vasomotor tone in the teat and an increased dilatation of the teat sphincter in these animals. PMID- 2560782 TI - Cutaneous metastasis from hepatocellular carcinoma resembling granuloma teleangiectaticum. AB - We reported a case of cutaneous metastasis originated from hepatocellular carcinoma, which appeared as a nodule resembling granuloma teleangiectaticum. The nodule, which was composed of tumor cells and intervening capillaries, showed characteristics similar to those of primary hepatocellular carcinoma. On immunohistochemical staining, the tumor cells were positive not only for AFP (alpha-fetoprotein), but also for CEA (carcinoembryonic antigen), which is negative in most cases of primary hepatocellular carcinoma. We discussed this case with a review of previous reports of skin metastasis from liver carcinoma. PMID- 2560783 TI - A case of skin metastasis of lung carcinoma associated with superior vena cava syndrome. AB - An unusual case of lung carcinoma with both skin metastasis and superior vena cava syndrome (SVCS) is reported. The histological type of the primary lesion as taken by punch biopsy was small cell carcinoma consisting of relatively small cells with hyperchromatic nuclei and scanty cytoplasm. The cutaneous metastatic lesion consisted of large cells with light-staining nuclei and small cells with deep-staining nuclei; it had foci of glandular elements by light microscopy. Neurosecretory granules characteristic of small cell carcinoma were found in the tumor cell cytoplasm by electron microscopy. Cutaneous metastatic rates, complication rates of SVCS, and histological varieties of small cell lung carcinomas are discussed. PMID- 2560784 TI - [Severe Aspergillus keratomycosis treated with itraconazole per os]. AB - A case of deep traumatic keratomycosis due to Aspergillus fumigatus with anterior chamber involvement is reported. Corneal perforation was threatening because of the large deep and long standing ulcer. This case emphasizes the difficulties of etiological diagnosis and treatment of keratomycosis. The authors analyse the peculiarities of corneal mycotic abcess and emphasize the importance of corneal cultures; they discuss the most recent therapeutic protocols for these lesions. After a very poor response to conventional antifungal therapy, total and quick recovery was acquired using itraconazole per os and topical Amphotericine B. The efficiency of itraconazole proves its antifungal activity against Aspergillus fumigatus and its good penetration to the deeper layers of the cornea and of the anterior chamber. PMID- 2560785 TI - Role of K+ ion channels in lymphokine-activated killer (LAK) cell lytic function. AB - Cells of the immune system possess K+ ion channels which have been implicated in various cellular functions including activation, differentiation and cytolytic function. To define the role of K+ ion channels in the lytic function of lymphokine-activated killer (LAK) cells, we investigated the effects of K+ channel blockers on their cytolytic activity. Results show that when LAK cell mediated cytolysis of AKIL-20 tumor cells was carried out in the presence of: a) the K+ channel blocker, 4-aminopyridine (4-AP); b) the monoamine, serotonin (5 hydroxytryptamine; 5-HT); c) the serotonin agonist, quipazine; d) or the Ca++ dependent K+ channel blocker, quinidine, the cytolytic activity of the LAK cells was inhibited in a dose-dependent manner. Preincubation of LAK effector cells also inhibited lysis in a dose-dependent manner, whereas preincubation of the AKIL-20 tumor target cells produced no inhibitory effects. This study demonstrates that K+ ion channels are involved in the LAK cell cytolytic process and that compounds, including neuroendocrine products, which modulate K+ ion channel function are capable of modulating the lytic activity of these effector cells. PMID- 2560786 TI - Imuthiol influences on cytotoxic T cells and NK activity in +/+ and athymic nude BALB/c mice. AB - The effects of imuthiol (sodium ditiocarb, DTC) on the expression of cytotoxic responses (CTL) and natural killer (NK) activity were evaluated in aged and young euthymic mice, and in nu/nu BALB/c mice. Imuthiol generated CTL and concomitantly reduced NK activity in nu/nu mice, suggesting that the agent can generate T cells in athymic nude animals. Treatment for up to 4 months augmented spleen NK and CTL activities in young or aged euthymic mice, but the generation of CTL in old animals was increased by long-term treatments better than by a single injection. The capacity of imuthiol to activate specific and nonspecific cytotoxic functions in euthymic mice may contribute to enhancement of resistance in vivo against transformed cells after treatment with this agent. PMID- 2560787 TI - Multiple pathways of angiotensin production in the blood vessel wall: evidence, possibilities and hypotheses. AB - In summary, multiple pathways of angiotensin production may exist in the blood vessel wall (Fig. 1), in addition to the well described renin--ACE enzymatic axis. It is not known whether these enzymatic pathways represent in vitro phenomena, or authentic in vivo alternate pathways which are activated only when the renin--ACE pathway is blocked, or whether they are operative at all times in the vessel wall. If these multiple pathways are functional, then the vascular angiotensin system may be very complicated, and may vary in different pathophysiological states. Future research in this area is likely to yield important and novel information on the in vivo pathways of production and function of vascular angiotensin. PMID- 2560788 TI - Effect of enalapril on aortic smooth muscle cell polyploidy in the spontaneously hypertensive rat. AB - The angiotensin converting enzyme (ACE) inhibitor enalapril was used to examine the effects of inhibition, regression and redevelopment of hypertension on the ploidy of aortic smooth muscle cells in spontaneously hypertensive rats (SHR). The incidence of polyploidy cells, as determined by flow cytometric DNA analysis, directly paralleled changes in systolic blood pressure. When the development of hypertension was inhibited by treatment with enalapril, the incidence of polyploid cells remained low compared with untreated age-matched SHR. Likewise, when the blood pressure of hypertensive animals was lowered by enalapril treatment, the incidence of polyploid cells decreased. Addition of enalapril to primary cultures of smooth muscle had no direct effect on proliferation or the incidence of polyploidy. These results suggest that angiotensin II may be involved in the development of vascular smooth muscle polyploidy in vivo, either by a direct effect on the cells or indirectly by elevating blood pressure or by potentiation of sympathetic discharge. PMID- 2560789 TI - Relationship of CA 125 and CA 19.9 with lung carcinoma histological subtype: preliminary study. AB - The aim of this work was to study the possible utility of simultaneous determination of CA 125 and CA 19.9 in patients with lung cancer. Serum levels of both markers were studied in 87 patients without metastases (Mo), 72 patients with distant metastases (MT) and 15 cases without clinical evidence of disease after primary treatment (NED). Sixty-five tumors were epidermoid, 34 were adenocarcinomas, 24 were cell undifferentiated carcinomas and 51 were small-cell carcinomas. Sera from 75 healthy subjects and 20 patients with benign lung disease were used as controls. The cut-off values used were 35 and 37 U/ml for CA 125 and CA 19.9, respectively. CA 125 and CA 19.9 serum levels were within normal limits in all control patients. In NED patients these markers were not elevated, except in one with chronic liver disease who showed elevated CA 19.9 (76 U/ml). Twenty-five percent of Mo lung cancer patients and 40.3% of MT cases had CA 19.9 over 37 U/ml. Abnormally high levels of CA 125 were found in 18.7% and 22.9% of Mo and MT patients, respectively. Sixty percent of patients with large cell undifferentiated carcinoma had elevated CA 125 (mean 176 U/ml) compared to 15.4% of patients with all other histological types of tumors combined (54.3 U/ml, p less than 0.01). CA 19.9 serum levels were also more often elevated in patients with large cell undifferentiated carcinomas (50%, 7/14 cases) than in other histological types (30%, 36/120 patients), but the difference was not statistically significant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2560790 TI - Varied cytochrome oxidase activities of the alveolar type I, type II and type III cells in rat lungs: quantitative cytochemistry. AB - Cytochemically demonstrated cytochrome oxidase activities in mitochondria of rat pulmonary alveolar epithelial cells were quantitatively compared. As the standardized procedures, lungs were fixed for 10 min at 4 degrees C with 2% pure glutaraldehyde, and incubated for different times at 37 degrees C in a medium containing 1.0 mg/ml 3,3'-diaminobenzidine-4HCl, 1 mg/ml cytochrome c, 0.1 mg/ml catalase, 7% sucrose, and 0.1 M phosphate buffer, pH 7.4. Electron micrographs were then obtained, and the densities of the reaction deposits in the mitochondrial intermembrane-intracristal space were measured in an image analyzer, and were plotted in terms of time in minutes. The initial velocity (maximal rate) of the activity expressed as deposit accumulation rate (% area/60 min) filling the mitochondrial intermembrane-intracristal space of Type I, Type II and Type III cells was 40, 130 and 9.3, respectively. These results indicated that mitochondria are varied in the intensity of cytochrome oxidase activity among 3 types of pulmonary alveolar epithelial cells, and that the image analyzing measurement of the accumulation rate of reaction product may be useful for quantitative analysis of enzyme activity, in particular, in the cells which are difficult to isolate from complex tissues. PMID- 2560791 TI - Polynucleotide cross-linking by aluminum. AB - The observations that there was an increased concentration of Al in the brains of Alzheimer's, Guam-Parkinson, and amyotrophic lateral sclerosis disease patients and that there was an apparent localization of the Al in chromatin led to a study of the interaction of Al(III) with DNA. We have previously shown that Al cross links calf thymus DNA at low pH (S. J. Karlik, G. L. Eichhorn, P. N. Lewis, and D. R. Crapper, Biochemistry 19, 5991 [1980]). Extended studies indicate that cross-linking occurs in DNAs of all base ratios, including polydAdT and polydGdC. Since Al cross-links prevent renaturation in polydAdT, the decrease in the amount of polymer renatured in the presence of Al becomes a quantitative appraisal of the extent of cross-linking. Saturation of cross-linking occurs at a 0.4 ratio of Al to nucleotide phosphate, indicating that potentially 80% of the base pairs are Al bound. Cross-links are broken at elevated pH and by EDTA. PMID- 2560792 TI - Lead chelates of meso- and racemic-dimercaptosuccinic acid. AB - Lead chelates of racemic- and meso-dimercaptosuccinic acid (DMSA) were synthesized and isolated from aqueous solutions and characterized by potentiometric measurements and infrared spectroscopy. Two types of lead chelates of racemic DMSA were isolated: one in which racemic-DMSA is coordinated to Pb+2 via one oxygen and one sulfur atom and the other in which the Pb2+ is coordinated via two sulfur atoms. The latter form of the chelate is converted into the former upon dissolution in dimethylsulfoxide. Only one type of Pb2+ chelate of the meso form of the ligand was formed. In this case, meso-DMSA is coordinated to Pb+2 via one oxygen and one sulfur atom. Meso- and racemic-DMSA have very different solubilities in aqueous solutions. Meso-DMSA is slightly soluble in water, whereas racemic-DMSA is very soluble in water even in the presence of strong acids. The solubilities of the chelates were found to be pH dependent. When the uncoordinated sulfhydryl and carboxylic acid groups dissociate, the chelates dissolve and remain in aqueous solution. The infrared spectra of meso- and racemic-DMSA show distinct features that can be used to detect the presence of either diastereoisomer. PMID- 2560793 TI - Genetic and physiological analysis of azole sensitivity in Saccharomyces cerevisiae. AB - Ketoconazole and fluconazole are azole antifungal agents which inhibit cytochrome P-450 mediated sterol C14 demethylation during ergosterol biosynthesis. We report on the activity of these antifungals on a variety of Saccharomyces cerevisiae strains grown under differing conditions known to affect cyt P-450 levels. Only slight increases in resistance to azoles were observed under conditions which induce the yeast cyt P-450 from undetectable levels. Strain variation was observed, with some strains exhibiting a fungicidal, and others a fungistatic response. Two cyt P-450 deficient mutants examined exhibited resistance to treatment with fluconazole and ketoconazole. This was attributed, at least in part, to an additional defect in sterol delta 5,6 desaturation and possibly to reduced cellular levels of azole drug. PMID- 2560794 TI - Interleukin 2 induced non MHC-restricted killing of herpes simplex type-1 (HSV-1) infected allogeneic and autologous lymphoblasts. AB - Peripheral blood lymphocytes (PBL) from herpes simplex type-1 (HSV-1) seropositive (sero+) and seronegative (sero-) individuals were assessed before and after in vitro culture with IL-2 for their ability to kill different tumor cells and to kill HSV-1 infected PHA lymphoblasts using a 4 hr 51Cr release assay. Fresh PBL from either sero+ or sero- individuals had little cytotoxic activity against HSV-1 infected autologous PHA lymphoblasts or against different NK-insensitive tumor targets. When PBL from either sero+ or sero- individuals were incubated 3-4 days in vitro with IL-2, significant levels of cytotoxic activity was observed against both allogeneic and autologous HSV-1 infected targets and against different NK-insensitive tumor cells. When comparing PBL from sero+ versus sero- individuals for IL-2 induced cytotoxic activity against HSV-1 infected autologous targets, PBL from sero+ individuals always had significantly higher levels of cytotoxic activity. Allogeneic HSV-1 infected targets were also killed by IL-2 activated PBL obtained from either sero+ or sero- individuals, but IL-2 activated PBL from sero+ individuals showed higher cytotoxic activity on HSV 1 infected autologous targets when compared to HSV-1 infected allogeneic targets. When IL-2 induced cytotoxic effector cells were treated with HSV-1 for 1 hr and recultured with IL-2, significant inhibition of cytotoxic activity against all target cells was observed. Greater than 50% inhibition of cytotoxic activity was observed when IL-2 induced cytotoxic effector cells were treated with HSV-1 and cultured for 6 hr.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2560795 TI - A critical review of NMR imaging and spectroscopy for the evaluation of cardiac hypertrophy in humans and experimental animals. AB - Nuclear magnetic resonance (NMR) is a product of modern technology and of great value for the noninvasive evaluation of cardiac hypertrophy. Among image diagnosis methodologies, 1H-magnetic resonance imaging is clinically available for exact visualization of the hypertrophic portion in the ventricular wall or septum. 31P-magnetic resonance spectroscopy is also promising for the evaluation of muscle energetics or intracellular environments. Various pitfalls that might be encountered in clinical or basic studies with NMR and desirable directions to be developed are described to make the modality more feasible for the evaluation of cardiac hypertrophy in humans and/or experimental animals. PMID- 2560796 TI - Role of adrenergic receptor systems in canine left ventricular hypertrophy. AB - Although the sympathetic nervous system and catecholamines have been postulated to play an important role in the development of myocardial hypertrophy, the precise mechanism is still ill-defined. We therefore investigated myocardial norepinephrine and the adrenergic receptor systems in two experimental canine models for cardiac hypertrophy; in 12 dogs with surgical cardiac denervation, and in 12 dogs with chronic infusion of a subhypertensive dose of norepinephrine at a rate of 0.04 mg/kg/day. After two months both models induced myocardial hypertrophy, indicated by significant increases in the heart weight, left ventricular wall thickness and cell diameter, as compared with 14 sham-operated control dogs. Cardiac denervation remarkably depleted myocardial norepinephrine while plasma norepinephrine remained unchanged. Both alpha 1- and beta-receptors were up-regulated, with Bmax increasing by 124% and 49%, respectively. The decrease in myocardial cyclic AMP content was relatively small as compared with the marked depletion in myocardial norepinephrine, probably compensated by augmentation of beta-receptor system activity. Chronic norepinephrine infusion also reduced myocardial norepinephrine content possibly due to stimulation of presynaptic alpha 2-receptor inhibiting norepinephrine synthesis and release. The number of alpha 1- and beta-receptors also increased by 97% and 30%, respectively, while myocardial cyclic AMP content remained unchanged. These observations indicate that neither direct stimulation of norepinephrine on the myocardial cell nor increased cyclic AMP is the mechanism for cardiac hypertrophy. A greater increase in the alpha 1-receptor, rather than in the beta receptors, in both models implies that a disproportional augmentation of the alpha 1-receptor system may play an important role in the development of myocardial hypertrophy. PMID- 2560797 TI - Signal and adaptational changes in gene expression during cardiac overload. AB - Chronic cardiac overload stimulates various quantitative and qualitative mechanisms of adaptation, some of them being species-specific. The signals responsible for these changes in gene expression are still speculative, nevertheless early modifications of the microtubular network have been reported. Soon after overload an increased expression of various genes coding for regulatory proteins has also been observed, this includes various oncogenes and the genes of several heat-shock proteins. Hypertrophy only, is non species specific and is adaptational because it both multiples the number of contractile units and it lowers wall stress. The slowing of the shortening velocity allows the heart to produce normal tension, at a lower cost, and has different biological explanations depending on the species. In small rodent ventricles, the main but probably not the unique, determinant of this physiological parameter is an isomyosin shift from a high ATPase activity form V1 to a low activity form V3, discovered in our laboratory in 1979. This shift has a transcriptional origin and also occurs in atria in every mammalian including humans; nevertheless it has not been evidenced in the ventricles of humans, dog, cat or guinea-pig. In these species it is necessary to take into account other mechanisms, namely those involved intracellular calcium movements. The number of total, and possibly active, calcium channels is normal in rat overloaded heart suggesting that their synthesis is activated commensurate to the development of hypertrophy. The situation is more complex for other sarcolemma proteins such as the beta adrenergic system and the Na+, K(+)-ATPase. For the latter there is presently some evidence that an isoenzymatic shift is likely to occur, at least in rats. PMID- 2560798 TI - Transcription of early developmental isogenes in cardiac myocyte hypertrophy. AB - We have developed a cell culture system to study molecular mechanisms important in myocardial hypertrophy. alpha 1-Adrenergic receptor stimulation produces hypertrophy of neonatal rat cardiac myocytes. Myocyte hyperplasia is not induced by alpha 1 stimulation, although alpha 1-adrenergic receptor-mediated DNA synthesis and cell division have been observed in other types of cells. The myocyte hypertrophic response does not require contractile activity. Activation of the alpha 1 receptor also produces highly specific alterations in gene expression, as measured at the mRNA and protein levels. In particular, there is selective up-regulation of two contractile protein isogenes that are expressed in vivo during early development and in pressure-load hypertrophy, skeletal alpha actin and beta-myosin heavy chain. Studies with an in vitro transcription assay indicate that stimulation of the alpha 1-adrenergic receptor leads to a distinctive temporal sequence of transcriptional activation. Transcription of the skeletal alpha-actin isogene is induced preferentially to that of cardiac alpha actin. Thus, early developmental isogene induction in alpha 1-stimulated hypertrophy reflects a fundamental change in the transcriptional program of the cardiac myocyte nucleus. The goal now is to define an intracellular pathway connecting the alpha 1-adrenergic receptor in the plasma membrane to activation of RNA polymerase II on the skeletal alpha-actin gene in the cardiac myocyte nucleus. There is evidence that protein kinase C may be one component of this pathway. A model for alpha 1-mediated transcription is presented. PMID- 2560799 TI - [Plasmapheresis and hormone therapy in severe forms of Guillain-Barre syndrome]. AB - Patients with severe Guillain-Barre syndrome were entered into 2 groups: group I was treated with hormones alone, group II with hormones plus plasmapheresis. The results were not significantly different. In view of recognized effectiveness of plasmapheresis for treatment of the syndrome, corticosteroid addition is inappropriate. PMID- 2560800 TI - Incidence of knee injuries and the need for further care. A one-year prospective follow-up study. AB - All visits to physicians paid due to knee injuries in the Orivesi Region Federation of Municipalities for Public Health Work in Finland were prospectively recorded over a period of one year. The visits totalled 360, which accounted for 1.0% of all visits. The number of patients was 148, and the total incidence rate of knee injuries was 1.1% corresponding to that during one year period every hundredth inhabitant sought medical treatment from a physician because of a knee injury. Forty-eight per cent of the knee injuries were sustained in sports activities. The injuries affected mostly knee ligaments and/or menisci. Every tenth visit led to a surgical consultation for assessment of the need for surgery, equalling three patients per 1000 inhabitants annually. The study provides basic information concerning the occurrence of knee injuries and the need for further care. The results can be used as an aid in the planning of arthroscopy as well as other examination and treatment resources of knee injuries as well as in the assessment of need for such services. PMID- 2560801 TI - Proton-decoupled, Overhauser-enhanced, spatially localized carbon-13 spectroscopy in humans. AB - Spatially localized, natural abundance, carbon (13C) NMR spectroscopy has been combined with proton (1H) decoupling and nuclear Overhauser enhancement to improve 13C sensitivity up to five-fold in the human leg, liver, and heart. Broadhand-decoupled 13C spectra were acquired in 1 s to 17 min with a conventional 1.5-T imaging/spectroscopy system, an auxiliary 1H decoupler, an air cooled dual-coil coplanar surface probe, and both depth-resolved surface coil spectroscopy (DRESS) and one-dimensional phase-encoding gradient NMR pulse sequences. The surface coil probe comprised circular and figure-eight-shaped coils to eliminate problems with mutual coupling of coils at high decoupling power levels applied during 13C reception. Peak decoupler RF power deposition in tissue was computed numerically from electromagnetic theory assuming a semi infinite plane of uniform biological conductor. Peak values at the surface were calculated at 4 to 6 W/kg in any gram of tissue for each watt of decoupler power input excluding all coil and cable losses, warning of potential local RF heating problems in these and related experiments. The average power deposition was about 9 mW/kg per watt input, which should present no systemic hazard. At 3 W input, human 13C spectra were decoupled to a depth of about 5 cm while some Overhauser enhancement was sustained up to about 3 cm depth, without ill effect. The observation of glycogen in localized natural abundance 13C spectra of heart and muscle suggests that metabolites in the citric acid cycle should be observable noninvasively using 13C-labeled substrates. PMID- 2560802 TI - Phase-distortion-free in vivo surface coil proton spectroscopy. AB - A 1-11 water suppression pulse sequence for phase-distortion-free surface coil NMR spectroscopy was constructed and tested in vivo on cat brain. The sequence produces brain spectra free of phase distortions, but some degradation of water suppression quality compared to closely related sequences which produce phase distortions was observed. PMID- 2560803 TI - [Fly ash and its biological effects. 3. Exposure to dust of workers in the energy generating industry (power plants and thermoelectric power stations)]. AB - An evaluation of exposure to dust of workers employed at typical work-stands in power industry plants was made. Mean concentrations of respirable dust determined at 14 different work-posts range from 0.45 to 8.95 mg/m3, and of total dust from 1.55 to 85.0 mg/m3. Mean content of free crystalline silica in dust samples is less than 10%. At the work-stands where ash dust was encountered, the presence of respirable fibres at concentration below 0.2 fibre/cm3 was observed. In all ash samples alpha-quartz and mullite were found: in some of them also kaolinite and orthoclase were traced. Only at five out of all 14 work-stands examined mean concentration of respirable dust was lower than the hygienic standard value. PMID- 2560804 TI - Protein kinase C mediates the effect of vasopressin in pituitary corticotrophs. AB - The role of protein kinase C (PKC) on vasopressin (VP) action was investigated by inhibition of endogenous PKC using prolonged incubation of the cells with phorbol ester, and by direct measurement of PKC activity in pituitary cells. Preincubation of the cells for 6 h with 100 nM TPA at 37 C resulted in a 90% decrease in total PKC activity. In the PKC-depleted cells, cAMP responses to stimulation with 100 nM CRF for 30 min were normal, but the potentiating effects of VP and PMA on CRF-stimulated cAMP production were abolished. The stimulation of ACTH secretion by VP and PMA alone was also abolished in PKC- depleted cells. PKC activity in cytosolic and detergent-solubilized membrane fractions from enriched pituitary corticotrophs obtained by centrifugal elutriation, was directly measured by enzymatic assays and by immunoblotting techniques. Basal PKC activity was higher in the cytosol than in the membranes (8.43 +/- 0.47 and 1.93 +/- 0.11 pmol 32P incorporated/10 min, respectively). After incubation of the cells with VP for 15 min or [3H] phorbol-12-myristate-13-acetate (PMA) for 30 min, PKC activity in cytosol was decreased by 40% and 89%, respectively, while the activity in the membrane was increased by 138% and 405%, respectively. Such VP- and PMA-induced translocation of PKC was also observed when the enzyme content in the cytosol and the membranes was measured by immunoblotting using a specific anti-PKC antibody and [125I]protein A. Autoradiographic analysis of immunoblots revealed an 80 kilodalton band characteristic of PKC, with OD higher in the cytosolic than in the membrane fractions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2560805 TI - Chloride channels mediate the response to gonadotropin-releasing hormone (GnRH) in Xenopus oocytes injected with rat anterior pituitary mRNA. AB - Functional expression of receptors for GnRH was studied using Xenopus laevis oocytes injected with poly(A)+ mRNA extracted from rat anterior pituitary glands. Whole-cell currents were monitored using two-electrode voltage-clamp techniques. In oocytes which responded to both GnRH and TRH, the GnRH response showed a longer latency and time-to-peak than the TRH response. The response to GnRH or an agonist of GnRH receptors, buserelin (1 nM-1 microM) consisted of current fluctuations and occurred in a dose-dependent manner. This GnRH response was blocked by the Cl- channel blockers 9-AC (9-anthracene carboxylic acid; 1 mM), 4,4'-diisothiocyanastilbene-2,2'-disulfonic acid (0.1 mM), and diphenylamine-2 carboxylic acid (0.1 mM). The reversal potential for the GnRH-induced current fluctuations was -25 mV, comparable with the reported Cl- equilibrium potential in Xenopus oocytes, and its shift, when the external concentration of Cl- was changed, was reasonably described by the Nernst equation. These results indicate that the GnRH-induced response was dependent on the activity of Cl- channels. Ca2+ also plays a role, as the GnRH-induced response was reversibly suppressed by a calmodulin inhibitor, chlordiazepoxide (0.2 microM), and by a blocker of intracellular Ca2+ release, TMB-8 (8-(N.N-diethylamino) octyl-3,4,5 trimethoxybenzoate; 0.1-0.2 mM). It is concluded that GnRH (and TRH) receptors, expressed in Xenopus oocytes by injecting exogenous mRNA from rat anterior pituitary glands, operate via activation of Ca2+-dependent Cl- channels. PMID- 2560806 TI - Mutational analysis of a lactogenic hormone reveals a role for lactogen-specific amino acid residues in receptor binding and mitogenic activity. AB - Mutant forms of mouse placental lactogen-II (PL-II) have been generated to assess the role of specific amino acid residues and protein regions in binding to the PRL receptor and in mitogenic activity. Conversion of any of three lactogen specific residues significantly reduced both of these hormone functions; mutation of the two other lactogen-specific amino acids revealed only minimal effects unless these changes were coupled with a second mutation. Deletions within the PL II protein all resulted in a complete loss of function, but switching regions between PL-II and proliferin, another member of the prolactin family in the mouse, did yield a chimeric protein with some PRL-like activity. This activity was increased substantially by conversion of one amino acid residue in the proliferin region to the corresponding lactogen-specific residue. The locations of the amino acids that have been found to affect hormone function are predicted to be closely apposed in the folded protein, suggesting that this region may be the site of interaction of this lactogenic hormone with the PRL receptor. PMID- 2560807 TI - Expression of recombinant human choriogonadotropin in Chinese hamster ovary glycosylation mutants. AB - Human CG, a member of the glycoprotein hormone family that includes LH, FSH, and TSH, is composed of two nonidentical subunits each containing two asparagine linked (N-linked) oligosaccharides. The role of the oligosaccharides in the action of these hormones is unclear. To examine the structure-activity relationships of the glycoprotein hormone oligosaccharides using nonenzymatic and nonchemical methods, we transfected CG subunit genes into mutant cell lines derived from Chinese hamster ovary cells. Two mutant cell lines that synthesize truncated oligosaccharides were used. Cell line 15B, lacking N acetylglucosaminyltransferase I, synthesizes N-linked carbohydrates containing Man5 oligomannosyl structures, and 1021, defective in transporting CMP-sialic acid into the Golgi, results in sialic-acid deficient glycoproteins. The binding of these derivatives to the LH/CG receptor did not differ significantly from purified CG (CR119), but the ability of the mutant hormones to stimulate cAMP biosynthesis in vitro is reduced compared to wild-type CG or CR119. Since the amino acid sequence of CG from the mutant and wild-type cells is identical, these data indicate that oligosaccharide structures, while not influencing receptor binding, directly affect signal transduction. PMID- 2560808 TI - The size heterogeneity of rat insulin-like growth factor-I mRNAs is due primarily to differences in the length of 3'-untranslated sequence. AB - Previous studies have revealed multiple size classes of rat insulin-like growth factor-I (IGF-I) of estimated size 7.5-7.0, 1.9-1.5, and 1.2-0.9 kilobases (kb). Available sequence information accounts for only 2.1 kb of the 7.5-7.0 kb IGF-I mRNAs. We used oligomer directed ribonuclease H (RNase H) mapping to define the extent to which the unknown sequence in the large molecular weight mRNAs lies 5' or 3' to known sequence. Rat liver polyadenylated RNAs were incubated with oligomer probes complementary to internal rat IGF-I precursor (E domain) coding sequences. RNase H was used to hydrolyze IGF-I mRNAs at the point of annealment with the oligomers. Resultant 5' and 3'-IGF-I mRNA fragments were analyzed on Northern blots. A probe specific for type 1 (class C) 5'-sequences (the most predominant of multiple 5'-sequence types found on rat IGF-I mRNAs) identifies intact IGF-I mRNAs of 7.5-7.0, 1.9-1.5 and 1.2-0.9 kb but, after oligomer directed RNase cleavage of these mRNAs, identified only a single IGF-I mRNA 5' fragment. Major differences in the length of sequence 5' to the IGF-I coding sequence therefore, do not account for the multiple size classes of type 1 (class C) IGF-I mRNAs. The size of the 5'-fragment suggests that the extent of sequence 5' to the IGF-I coding sequence is 0.4-0.7 kb in type 1 (class C) IGF-I mRNAs. Identification of multiple 3'-fragments of IGF-I mRNAs demonstrated heterogeneity in the 3'-ends of rat IGF-I mRNAs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2560809 TI - Human chorionic gonadotropin regulates expression of the proenkephalin gene in adult rat Leydig cells. AB - The gene encoding the proenkephalin precursor is expressed in both progenitor spermatogenic cells and somatic cells in the rat testis. The 1750-nucleotide (nt) germ cell-specific proenkephalin mRNA is the predominant form detected in the adult rat. The 1400-nt somatic cell form has previously been shown to be expressed in cultured Sertoli cells derived from sexually immature rats, and FSH treatment increases expression of the proenkephalin gene in that experimental system. In the present report we demonstrate that the 1400-nt proenkephalin transcript is present in a freshly isolated, enriched preparation of adult rat Leydig cells. This gene continues to be expressed when the Leydig cells are maintained in primary culture. Treatment of cultured Leydig cells with hCG or a cAMP analog leads to a rapid increase in proenkephalin-mRNA levels. The expression of the proenkephalin gene throughout rat development in the two major somatic cell types of the testis further suggests that proenkephalin-derived peptides play an important role in modulating testicular function. The gene encoding another of the opioid peptide precursors, POMC, has previously been reported to be expressed in rat Leydig cells. We were unable to detect POMC transcripts in cultured Leydig cells. POMC mRNA is abundant in a germ cell enriched cell fraction. PMID- 2560810 TI - Control of thyroperoxidase and thyroglobulin transcription by cAMP: evidence for distinct regulatory mechanisms. AB - The expression of the genes coding for thyroglobulin (TG), and thyroperoxidase (TPO), are regulated by TSH. These effects are mediated by cAMP as they are reproduced by forskolin. In vitro run-on transcription assays performed on nuclei isolated from dog thyrocytes in culture or from dog thyroid slices, indicate that the forskolin-induced transcriptional stimulation of TG and TPO genes are very different. For the TG gene, the kinetics of transcriptional activation vary according to the experimental model: it is rapid (1 h) in thyroid slices and slow (8 h) in primary cultures. In contrast, TPO induction is rapid in both cases. In primary cultures, insulin is responsible for the basal level and for a part of forskolin-induced TG transcription, whereas TPO transcription is not affected by insulin. The forskolin-induced increase of TG transcription requires ongoing protein synthesis, as it is blocked by cycloheximide. TPO gene transcription is unaffected by cycloheximide. Taken together with previous data on the two genes, our results suggest that while TPO regulation corresponds to the classical model of genes in which the promoter is regulated directly via cAMP regulatory elements, TG gene regulation involves the synthesis of an intermediary, rapid turnover trans-acting protein. PMID- 2560811 TI - Chronic stimulation of glucose transporter gene expression in L6 myocytes mediated via the insulin-like growth factor-1 receptor. AB - We have used differentiated L6 myocytes to investigate the regulation of glucose transporter gene expression by insulin and insulin-like growth factor-1 (IGF-1). Chronic exposure to insulin (1 microM) or IGF-1 (10 nm) resulted in a 2- to 5 fold stimulation of 3H-2-deoxy-D-glucose uptake and a corresponding increase in the expression of rat brain/HepG2-type glucose transporter mRNA (GTmRNA) and immunoreactive transporter protein. The dose responses to both insulin and IGF-1 for stimulation of glucose uptake were paralleled by the expression of GTmRNA. Glucose uptake and GTmRNA levels were half maximally stimulated by 350 and 100 nM insulin, respectively, or by 2 nM IGF-1. Comparison of receptor occupancy with stimulation of glucose uptake and GTmRNA expression suggests that insulin exerts its effects through the IGF-1 receptor. Fibroblast growth factor, epidermal growth factor, platelet-derived growth factor, and phorbol ester had little or no effect on GTmRNA expression. These results demonstrate that the IGF-1 receptor mediates chronic regulation of transporter mRNA expression and protein synthesis and activity in cultured rat muscle cells. PMID- 2560812 TI - [Isolation of recombinant vaccine strains of poliovirus from patients with poliomyelitis]. AB - Oligonucleotide maps of some poliovirus type 2 strains isolated from polio cases, while being clearly related to that of the Sabin vaccine type 2 strain, exhibited, nevertheless, marked differences from the reference (vaccine) map. Several large oligonucleotides derived from 4 such strains were subjected to enzymatic sequencing. The results strongly suggest that all of them were intertypic recombinants between the Sabin strains. The 5'-parts of the genomes of these strains were derived from the type 2 vaccine whereas the 3'-parts were of type 1 (in 2 strains) or type 3 (in other 2 strains) origin. PMID- 2560813 TI - [Biotinylated DNA probe for detection of streptotricin acetylation genes]. AB - The biotin-labelled DNA probe for identification of functioning and silent genes for streptotricin acetylation has been constructed. The probe is homologous to sat1 gene of the movable genetic element Tn1825. The simplified modification of the hybridization technique using the biotin-labelled DNA probe is described. PMID- 2560814 TI - [A new plasmid pBS1001 with broad range of hosts]. AB - A new broad host-range plasmid capable of conjugative transfer has been isolated and characterized. The plasmid has the high frequency of conjugation transfer, is capable of conjugative transfer mobilization of nonconjugative plasmids, carries no known phenotypic markers. The plasmid demonstrates the specific interaction with the plasmids of P incompatibility group. The comparatively small size of the plasmid permits one to use it efficiently for comparative study of organization of the broad host range plasmids. PMID- 2560815 TI - [The use of Pseudomonas sp. M plasmid pM3 for the transposon mutagenesis of Erwinia]. AB - The transposon containing derivatives pMTF9 (Tn9), pMTF10 (Tn10) and pMTF59 (Tn5, Tn9) of the Pseudomonas sp. M conjugative plasmid pM3 demonstrating temperature dependent instability in Erwinia cells incubated at 37 degrees C have been isolated. The obtained plasmids have been shown to be usable for transposon mediated mutability in the bacterial cells of Erwinia generum incubated at 37 degrees C. PMID- 2560816 TI - The varietal differences in some biological activities of proteins extracted from flours of wheat seeds harvested in 1986. AB - Six wheat seeds varieties taken for experiments showed some differences in crude protein content ranging from 10.4-12.8%. A procedure for protein fractionation by the successive elimination of albumins, globulins and gliadins from the wheat varieties studied was elaborated. In the obtained protein fractions endogenous amylolytic and inhibitory activities tested against bovine pancreas trypsin and alpha-amylases of mammalian and insect origin were determined. The determined biological activities differ considerable among wheat varieties studied and were mainly located in albumins and gliadins while only a small amount were detected in globulins. Extractable proteins were relatively thermostabile and powerful inhibitors of the alpha-amylases examined. As the influence of environmental conditions was the same for all wheat varieties studied the observed differences in biological activities should be rather attributed to the genetic differences. PMID- 2560817 TI - The microbiological quality of Erzincan (Savak) Tulum cheese from Turkish retail markets. AB - Twenty samples of Erzincan (Savak) Tulum cheese were investigated for the microbiological quality and some chemical analyses. Cheese was characterized by moisture content means = 45.0%; 3.27% sodium chloride and 2.14% acidity. Significant variation was found in the major compositional factors indicating a general lack of quality and/or extreme diversity of the manufacturing conditions. Microbiological test revealed the presence of very high count of total coliforms, psychrotrophic bacteria, yeasts and moulds, high numbers of Staphylococcus aureus and Clostridium perfringens but no salmonellas. Statistical relationship between growth and counts, and the presence of other indicators, pathogens, and compositional factors was investigated. PMID- 2560818 TI - Effect of naftidrofuryl on metabolism and survival of cultured neurons. AB - The direct influence of Naftidrofuryl, a drug widely used for the treatment of cerebrovascular diseases, on the metabolism and survival of neurons was investigated in 8-day-old chick embryo forebrain cultures. Interaction of Naftidrofuryl with neurons resulted in the increase of the intracellular cyclic AMP levels. Naftidrofuryl stimulated deoxyglucose uptake and lactate production in a time- and dose-dependent fashion. These effects were observed after a few minutes following the beginning of the treatment with Naftidrofuryl. In parallel to the action on the metabolic activities, Naftidrofuryl was able to increase the survival rate of a significant proportion of the neuronal population. These data support the notion that Naftidrofuryl may act as a neuroprotective agent. PMID- 2560819 TI - Characterization of a co-culture system of neurons and hepatocytes. AB - A co-culture system of cerebellar granule cells (glutamatergic neurons) and hepatocytes has been developed. Petri dishes divided in halves by a temporary septum were coated with poly-L-lysine and cerebellar granule cells plated in one of the compartments. Five days later hepatocytes were plated in the other compartment and after 2 days the septum was removed and the two cell types shared the same culture medium for a period of 5 days. During this period of time cultures of neurons and hepatocytes kept separately or in co-culture exhibited identical characteristics with regard to activities of pyruvate kinase and glucokinase (hepatocytes), aspartate aminotransferase (neurons) as well as evoked transmitter release (neurons) and content of cytochrome P-450 (hepatocytes). The results show that it is possible to maintain neurons and hepatocytes in co culture sharing the same culture medium for a prolonged period of time. Such a system may serve as a pharmacological model to study interactions between liver and brain cells with regard to neuroactive drugs. PMID- 2560820 TI - Cyclic nucleotide phosphodiesterase and 5'-nucleotidase: a coupled system. AB - Evidence is presented that multiple forms of cyclic nucleotide phophodiesterase (PDE) activity chromatographically separated from the soluble fraction of bovine hypothalamus are co-eluted with multiple forms of 5'-nucleotidase (5'N) activity. The enzymes could not be resolved from each other by anion-exchange chromatography on DEAE-TSK; by affinity chromatography on phenyl-, blue-, concanavalin A-, 5' AMP-sepharose, cAMP-silica gel; or by gel filtration on sephacryl S-200. The catalytic activities were found to be associated with the tetrameric, dimeric, and monomeric forms of the enzymes. The molecular weights determined by gel filtration or by SDS-gel electrophoresis were 220, 114, and 57 kDa, respectively. Kinetic analysis revealed that the first-order rate constant for 5'AMP hydrolysis measured in the reactions: cAMP----5'AMP----adenosine was 100 times higher than that in the reaction: 5'AMP----adenosine. Thus, functional interrelation between PDE and 5'N was expressed in drastic acceleration of the consecutive reactions: cAMP----5'AMP----adenosine. The results confirm the conclusion about the existence of a multienzyme system involving PDE and 5'N or of a single bifunctional enzyme in brain tissue. PMID- 2560822 TI - Testicular cancer. PMID- 2560821 TI - Naloxone-6 alpha-spirohydantoin: a long-acting opiate antagonist. AB - We have synthesized a novel derivative of naloxone, naloxyl-6 alpha spirohydantoin. The compound was rigorously purified and its stereochemical structure characterized. In tests in vivo in mice, it was less toxic than the parent compound and it showed weak anticonvulsive activity. It exerted antagonist effects comparable to those of naloxone but its effects were longer lasting, persisting for a week. PMID- 2560823 TI - Aniridia, Wilms' tumor and human chromosome 11. AB - Aniridia-a developmental abnormality of the eye in which the iris is apparently absent-has been shown to be genetically associated with Wilms' tumor (an embryonic nephroblastoma) in the WAGR syndrome. Genetic and cytogenetic evidence points to band p13 of human chromosome 11 as the localization of the genes responsible for these defects. Deleted chromosomes 11 from WAGR patients and clinically associated translocations involving 11p13 have been used to map and order genes and anonymous DNA markers around the WAGR locus refining the localization of the aniridia and Wilms' tumor genes to within about 1 million base pairs of DNA. PMID- 2560824 TI - Luteotrophic agents and steroid substrate utilization. AB - The cells of the CL are derived from precursors in the ovarian follicle. For their existence and function, they are dependent on a combination of various substances, collectively described as luteotrophins. These luteal stimulatory agents, including luteinizing hormone and prolactin from the pituitary, chorionic gonadotrophin from the placenta, and insulin from the pancreas, are classified as endocrine luteotrophins because they are produced at a distance from their target organ, the CL. Paracrine and autocrine luteotrophins are substances produced by the ovary with specific stimulatory effects on the CL: oestrogens and insulin like growth factors meet this criterion. The CL produces progesterone from cholesterol which can be synthesized de novo or be derived from cholesteryl esters stored in the CL or from free and esterified cholesterol from lipoproteins in circulation. Cholesterol appears to be transported within the luteal cell by means of the cytoskeleton, by a specific, labile intracellular protein and/or by a specific sterol carrier protein. In this review, we have presented evidence to indicate that the luteotrophins have effects on all forms of cholesterol available to the CL. A major mechanism of action of luteotrophins appears to be the management of cholesterol supplies. LH, acting through a cAMP second messenger system, initiates the process of progesterone synthesis by induction of the cleavage of the side-chain on free cholesterol to produce pregnenolone. LH also acts to hydrolyse cholesteryl esters to produce free cholesterol. HDL metabolism appears to be affected by LH, hCG and cAMP. There is good evidence to suggest that luteal cell binding of HDL is regulated, in part, by LH. All phases of the process of utilization of LDL-cholesterol, including the availability of the LDL receptors, internalization of the LDL receptors and ligands into the luteal cell and degradation of LDL, are regulated by LH and cAMP. These agents induce an increase in mRNA for the LDL receptor. These processes may be, in part, controlled by an intracellular negative feedback mechanism for which cholesterol is the effector. However, in experiments where intracellular cholesterol was elevated by hydroxylated cholesterol or by inhibition of the conversion of cholesterol to pregnenolone, it was demonstrated that LH can override the cholesterol negative feedback mechanism. PRL plays an active role in cholesterol homeostasis in the luteal cell. The available evidence suggests that it is not involved in cholesterol side-chain cleavage, and enhances rather than reverses the formation of cholesteryl esters.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2560825 TI - [The critical 3-day fever-exanthema in young children (exanthema subitum, Zahorsky roseola infantum)--what is new?]. AB - As to the present knowledge the critical rose rash of infants (exanthema subitum, roseola infantum) means to be an exanthematous infectious disease that, occurring preferably in elder babes and younger infants (1st--3rd year of life), is caused by the newly detected herpesvirus (now the sixth one) pathogenic for man. The natural contamination in our latitude is intense (60-75%), and the probably lifelong immunity in the majority of cases is acquired in infancy. Though experts in the clinical subject do stress all the time the exanthema subitum to be the most frequent exanthematous disease in early infancy and infancy, infection chains tested to exanthema, respectively epidemics are observed decidedly seldom. Consequently, most of all infections use to develop clinically inperceptibly or even with other symptoms (and without an exanthema); the exanthema up to now obligatory for establishing the diagnosis uses to appear only in the minority of all cases and in the great majority of them the seroconversion is clinically silent. The prognosis of the exanthema subitum is in force to be good; the disease is the special field of activity for ambulatorily acting physicians (paediatrist, general practitioner). To begin with, the status diagnostically unclear and high-febrile for several days, the central-nervous excitability occurring in many cases, and in some children the appearing of dramatic febrile convulsions, as well as gastroenteric symptoms perpetually give rise to differential diagnostic considerations and sometimes even to a (false) antibiotic therapy. In case of an affection by herpesviruses principally a latency (persistence) of the virus lasting for years (possibly even lasting for life) in human beings is to be taken into account; this condition is also current for HHV 6. On occurring of an immune debility a release and a discharge of herpesviruses- not only by children suffering from an exanthema subitum (!)--are possible so that human beings of each age may come into question to be the source of infection. Double infections of human immune cells (for instance HHV 6 and HIV 1 simultaneously) already have been found. The necessary studies in order to clarify essential clinical and virological problems are world-widely in full activity, and new cognitions (further symptoms of diseases associated with HHV 6, possibly affections in prenatal infections and so on) are soon to be taken into account. PMID- 2560826 TI - [The participation of opiatergic mechanisms in the regulation of bone marrow hematopoiesis in stress]. AB - It is shown that immobilization stress intensifies functional activity of hematopoietic precursor cells, which leads to the development of medullary hyperplasia. Injection of dalargin into immobilized animals as well as its addition to the culture medium in producing clones of myelokaryocytes derived from donors subjected to stress in vitro reduces the colony- and cluster-forming activity of bone marrow cells and causes suppression of erythro- and granulocytopoiesis. Naloxone abolishes the suppressive effect of dalargin in vivo and in vitro but at the same time attenuates bone marrow hyperplasia in stress. It is suggested that naloxone may block both the stimulating and the inhibiting effect of enkephalins on hematopoiesis in stress. PMID- 2560827 TI - [The endogenous opioid system in shock]. PMID- 2560828 TI - [Developmental disorders resembling Apert syndrome in a 2-year-old girl]. PMID- 2560829 TI - [Analysis of neoplasm metastases to the bone marrow in patients with lung cancer]. AB - The authors analysed bone marrow metastases in lung cancer in 104 deceased patients. Trepano-biopsy was taken from the sternum, hip bone and spine. Bone marrow metastases were found in 33 cases (31.73%). Most often they were seen in small cell lung cancer (16 cases--35.56%). In 12 cases the bone marrow was the only site of lung cancer metastases. PMID- 2560830 TI - [Evaluation of the second line of treatment of patients with small cell carcinoma of the lung]. AB - Efficacy of second line therapy which included vepesid and adriamycin or vepesid, adriamycin and cis-platin, in small cell lung cancer was studied. Objective remission was seen in 35% of the once more treated patients. Analysis of survival time showed significant prolongation of it in cases treated once more in comparison with a group of patients with progression of the neoplastic process during primary therapy or patients not treated due to relapses of the process. The observed difference was bigger in patients that reacted with a remission to the second line therapy. PMID- 2560831 TI - [Phosphoinositides and their role in the development of chronic complications of diabetes mellitus]. PMID- 2560832 TI - [Retroviruses with 2 oncogenes]. AB - Less than a decade ago, some acute transforming avian leukemia retroviruses that transduced not one but two distinct cellular genes that could cooperate in the viral-proned transformation processes was discovered. In this paper three examples of such retroviruses are presented. This are: avian erythroblastosis virus ES 4 (AEV-ES 4), avian erythroblastosis virus E 26 (AEV-E 26), avian Mill Hill-2 myelocytomatosis virus (MH 2). PMID- 2560833 TI - [Dental care in a temporary after-care unit for haemophiliacs]. AB - The author found within the framework of comprehensive treatment of haemophiliacs in the temporary after-care unit of the department of clinical haematology of the Regional Institute of National Health in Plzen that on average 58.5% of the patients have carious teeth. They present their experience with dental treatment of these patients and discuss some problems. They emphasize that systematic, early and careful dental care of haemophiliacs within the framework of care at the district level is essential. PMID- 2560834 TI - [Computerized tomographic diagnosis of hepatocarcinoma after injection of lipiodol into the hepatic artery]. AB - Iodized oil (lipiodol) injected into the hepatic artery is selectively retained by hepatocarcinomas, as demonstrated by computerized tomography (CT) performed one week after the injection. The value of this technique for the diagnosis of hepatocarcinoma was assessed in a retrospective study of 45 patients. In 39 per cent of the cases intrahepatic tumoral extension was determined by the iodized oil which showed tumoral nodules that had not been detected by conventional methods, such as ultrasonography and CT alone. The lesions revealed by the iodized oil were small nodules around the main tumour. The combined iodized oil CT technique plays an important role in the choice of treatment, especially when surgical excision is contemplated. It might also contribute to an early diagnosis of hepatocarcinoma in patients at risk, as illustrated by four of our cases where conventional morphological examinations had been negative. PMID- 2560835 TI - Action of antidepressants on the septal nuclei of the rat. AB - Studies on neuronal firing have shown a decrease in frequency of firing in structures not directly related to emotional processes. However, studies of the hippocampus have shown increases in firing rate. Other limbic structures not yet explored in regard to the action of antidepressants include the septal nuclei. The present work describes a common effect of various therapeutic antidepressant models. Extracellular unit recordings were obtained from the septal nuclei of rats exposed to different acute treatments: clomipramine, isocarboxazid, trazodone, sleep deprivation, and electroshock. Frequencies and firing intervals were analyzed. After treatment, an increase in firing frequency in cells of the dorsolateral septal nucleus was found. This supports the hypothesis that brain structures related to the phenomenon of self-stimulation participate in the mechanism of antidepressant treatments. PMID- 2560836 TI - [Human papillomaviruses and malignant neoplasms]. PMID- 2560837 TI - [Variability of the clinical picture and the course of infection of the genital organs an anus by human papillomaviruses]. PMID- 2560838 TI - Comparison of intra-arterial digital subtraction angiography with conventional angiography after transcatheter arterial embolization for hepatic malignancy. AB - Twenty patients with hepatic malignancy were treated with transcatheter arterial embolization (TAE) under examination by intra-arterial digital subtraction angiography (IA-DSA) and conventional angiography (CA). Comparison of these two angiographies revealed that the time required for confirmation of the embolized portion of the artery was about four-and-a-half times shorter with IA-DSA than CA. Moreover, IA-DSA revealed the obstructed portion more readily and accurately than CA. In particular, confirmation could not be made by CA in 35% of cases because of residual Gelform sponge containing contrast medium in the artery. The visualization of residual tumor stain after TAE was 40% better on IA-DSA than CA, because of residual Gelform sponge containing contrast medium and overlapping shadow with calcified costal cartilage, excretory pyelography, and original tumor stain. However, CA was better than IA-DSA in visualizing accidental obstruction of nonobjective arteries such as the cystic artery because of the poor spatial resolution and misregistration artifacts of IA-DSA. CA was better than IA-DSA in visualizing surrounding nonembolized hepatic parenchyma because of the misregistration artifacts of IA-DSA. For effective TAE without severe complication, we concluded that TAE for HCC should be performed under a combination of IA-DSA and CA. PMID- 2560839 TI - [Mitochondrial encephalomyopathy]. AB - Modern concepts regarding mitochondrial encephalomyopathies (ME) are summarized. Utilizing recent techniques of molecular biology we studied some cases of ME referred to the Institute of Clinical Neurology of Milan University. With these techniques we demonstrated different mitochondrial DNA deletions either in patients' muscle or in culture. PMID- 2560840 TI - [In vitro susceptibility of Cryptococcus strains to 5 antifungal drugs]. AB - A comparative study of the "in vitro" susceptibility of 24 Cryptococcus strains to 5 antifungal drugs (amphotericin B, 5 fluorocytosine, miconazole, itraconazole and ketoconazole), was carried out. These strains were grouped according to species, varieties and isolation's origins. The minimum inhibitory concentration (M.I.C.) was determined by the agar dilution technique in yeast nitrogen base agar with dextrose. The mean geometrical of the M.I.C. values of each group was compared with the others. The results obtained were homogeneous with the only exception of the "non neoformans" strains, in which, higher M.I.C. to 5 fluorocytosine values were detected. PMID- 2560841 TI - Fifth disease in children living in Belem, Brazil. AB - Acute sera from two children suffering from an illness with an erythematous rash were positive for B 19 virus specific IgM antibody, as tested by a capture radioimmunoassay. The first patient, a two year old boy, presented with a cutaneous rash of six days duration, the second was a four year old girl, sister of the first patient, who was examined at the same time and had a three day history of cutaneous rash. PMID- 2560842 TI - Fecundability and husband's age. AB - The effect of husband's age on the probability of conception is evaluated from World Fertility Survey data in five developing countries: the Ivory Coast, Ghana Kenya, the Sudan, and Syria. Proportional hazards models, which include wife's age, husband's age, marriage duration, union type, and post-partum exposure as covariates, are used to describe the monthly conception rate for second and higher-order birth intervals in which no contraception was used. With the exception of Syria, the resulting models indicate that the effects of male age are generally small in relation to the influences of marital duration and the age of the woman. PMID- 2560843 TI - [The complex assessment of local inflammation in rheumatoid arthritis patients]. AB - Application of a complex of clinical, thermography and radionuclide methods in 79 patients with rheumatoid arthritis has demonstrated high sensitivity of scintigraphy with the use of osteotropic radiopharmaceuticals (99mTc pyrophosphate, 99mTc-phosphone) as well as a possibility of diagnosing preclinical injuries to the joints. The thermography and scintigraphic parameters depended on the degree of inflammation, permitting an objective evaluation of the treatment effect. PMID- 2560844 TI - [Beta-endorphins and bronchial asthma in children]. AB - The content of beta-endorphins was studied in 64 children aged 6 to 14 years with atopic bronchial asthma (BA). The phasic relationship was established in the time course of changes in the content of beta-endorphins. In the paroxysmal period of BA, the concentration of beta-endorphins in blood was lowered, whereas after attacks and in the stage of remission, the level of beta-endorphins was increased and differed significantly from the indicators in the control group. The reduction of the content of beta-endorphins during exacerbation correlated with the disease gravity. The content of endorphins was found to be affected by the age and sex: the greater reduction of the level of beta-endorphins was recorded in children aged 6 to 11 years and in boys. The reduction of the level of beta endorphins was followed by the impairment of interhormonal relations between ACTH and beta-endorphins, ACTH and hydrocortisone in all the disease phases. A correlation was discovered between the level of beta-endorphins and ACTH, cAMP as well as the level of IgE in blood plasma. It is suggested that derangements in the endorphin system may influence the mechanisms by which bronchial hyperreactivity is formed. PMID- 2560845 TI - Electron microscope immunolocation of gap junctions in Drosophila. AB - Gap junction immunolocation was carried out in thin sections of Lowicryl K4M embedded Drosophila imaginal wing discs using an affinity-purified polycolonal anti-18kD gap junction protein antibody and a colloidal gold-conjugated secondary antibody. Colloidal gold labelling was predominantly associated with obliquely sectioned gap junctions, the ends of junctional profiles and other regions in which the adjacent junctional membranes were separated or distorted. The pattern of staining suggests that the determinant recognized by the antibody is relatively inaccessible, probably with a topological location in the transmembrane or extracellular domain of the membrane-spanning connexin protein. PMID- 2560846 TI - An extract of lionfish (Pterois volitans) spine tissue contains acetylcholine and a toxin that affects neuromuscular transmission. AB - A soluble toxic extract derived from spine tissue of the lionfish (Pterois volitans) decreased heart rate and force of contraction in isolated clam and frog hearts. These actions were due to the presence of micromolar concentrations of acetylcholine in the extract. Toxicity was retained after hydrolysis of acetylcholine by exogenous acetylcholinesterase, but heart function was no longer affected. Toxin treated in this way induced muscle fibrillation in an isolated nerve-muscle preparation, followed by blockade of neuromuscular transmission. Bursts of transient depolarizations were recorded at the muscle endplate shortly after toxin addition that correlated in time with the duration of toxin-induced muscle fibrillation. These effects are thought to be due to the increased release and then depletion of acetylcholine from the nerve terminal. PMID- 2560847 TI - [The possibility of preparing a live vaccine against tick-borne encephalitis from Malay langat virus TP-21]. PMID- 2560848 TI - [The significance of humoral immunity in post-vaccination resistance of mice to tick-borne encephalitis]. PMID- 2560849 TI - [Laboratory methods of controlling immunogenicity of vaccines against tick-borne encephalitis]. PMID- 2560850 TI - Bovine leukemia virus--issue and model. A satellite meeting to the XIIIth symposium of the International Association for Comparative Research on Leukemia and Related Diseases. Eilat, Israel, November 1987. Proceedings. PMID- 2560851 TI - Bovine leukemia virus--issue and model. PMID- 2560852 TI - Expression of the bovine leukemia virus transactivator protein p34 by a recombinant vaccinia virus. AB - In order to characterize the bovine leukemia virus transactivator protein, a recombinant vaccinia virus (v-LOR) containing the BLV post-envelope long open reading frame was constructed. v-LOR was shown to encode a functional protein able to transactivate the BLV long terminal repeat-directed gene expression in the infected cells. The encountered level of transactivation was about one third of that measured in BLV-infected fetal lamb kidney cell lysates. PMID- 2560853 TI - Tumor-associated antigen on bovine leukemia virus-induced bovine lymphosarcoma. AB - Specific tumor-associated antigen (TAA) was detected on enzootic bovine leukosis (EBL) cells by monoclonal antibodies against TAA. One of the monoclonal antibodies, c143, reacted with all EBL tumor cells tested but not with bovine leukemia virus (BLV) antigens. c143 reacted slightly with bovine fetal thymus and mitogen-stimulated lymphocytes from BLV-free cows but not with normal bovine lymphoid cells. TAA may be a good tumor marker of EBL tumor cells. We sacrificed eight TAA-positive but clinically normal animals and examined them in order to elucidate whether or not they had gross or histological tumors. At necropsy, four animals had tumors macroscopically. Three animals had no tumors histologically but had initial lesions showing follicular hyperplasia and the TAA on affected lymph nodes. The one remaining showed medullary hyperplasia in the spleen but there were no findings of tumors. Thus, c143 is a useful tool not only for diagnosing EBL, but also for screening of BLV-infected cattle with potential to develop tumors in the future. PMID- 2560854 TI - Immunohistological demonstration of virus and tumor associated antigens in tissues experimental and spontaneous bovine leukemia virus (BLV) infection. AB - Expression of bovine leukemia virus (BLV) antigens in vivo has not been shown. After BLV infection, however, production of antibodies directed towards BLV proteins (e.g. gp51) can be easily demonstrated. Thus, production of BLV proteins has to take place somewhere in infected cattle. Tissues and organs of experimentally infected cattle were fixed in acetone and embedded in paraffin. Monoclonal antibodies directed to gp51 were used to demonstrate BLV expression immunohistologically by the peroxidase-antiperoxidase (PAP) method. The same samples were also used to demonstrate a tumor associated antigen (TAA) employing a monoclonal antibody. Our results indicate that very few cells, found in the intestinal mucosa, produce gp51 in vivo. The expression of TAA, however, increases significantly shortly after infection with BLV and remains high throughout life. PMID- 2560855 TI - The BLV-induced leukemia--lymphosarcoma complex in sheep. AB - Sheep are highly susceptible to BLV infection and can be infected via several different means (routes). In all inoculated animals, specific anti-BLV antibodies can be demonstrated 1 to 3 months post-inoculation (p.i.). Between 10 and 13 months p.i., a moderate but persistent lymphocytosis (PL) may be detected in about 50% of the infected animals. This hematological disorder may be, but is not necessarily, associated with the development of a lymphosarcoma and can (might) be interpreted as a true lymphoid leukemia. According to findings revealed by immunolabelling and mitogen stimulation of peripheral blood lymphocytes, BLV induced PL appears to be a B-cell disorder. Induced lymphosarcoma appears in about 40% of infected sheep during the 6 years p.i. It too is of B-lymphocyte lineage. In vitro studies demonstrate that BLV antigen is expressed exclusively in B-lymphocytes. Yet, BLV expression is greatly stimulated in whole lymphocyte culture by the addition of T-cell mitogen. This same phenomenon occurs when the supernatant of stimulated T-lymphocyte cultures is added to isolated BLV-infected B-lymphocytes. This observation supports the hypothesis that, as is the case with other retroviruses such as HIV, BLV is able to use the regular activation machinery of the immune system for its own replication and transmission. It seems, therefore, that the leukemia-lymphoma complex in sheep may serve as an accurate experimental model for the study of the biological properties of retroviruses. PMID- 2560856 TI - Chemotherapy and immunotherapy of bovine leukosis. AB - To prevent the progression of the disease, we treated leukemic or preleukemic cows with (1) adriamycin (ADM) entrapped in liposomes conjugated with monoclonal antibody, c143, against tumor-associated antigens (TAA) of bovine leukemic cells and (2) an immunotherapy using an immunopotentiator consisting of the cell wall skeleton of Nocardia rubra (N-CWS). Five leukemic or preleukemic cows with TAA positive peripheral blood lymphocytes (PBL) received four injections of ADM alone (0.4 mg/kg body weight) or c143-conjugated liposomes containing the same dose of ADM (L-ADM-c143) through the jugular vein at about 4-day intervals. In three animals treated with L-ADM-c143, the TAA-positive cells gradually decreased with treatment and finally two animals became TAA-negative during a 6-week period and a 14-week period after treatment, respectively. About 6 weeks later, however, TAA positive cells gradually increased. In the control, two animals treated with ADM alone showed no decrease of TAA-positive cells. Five TAA-positive animals with enlarged subcutaneous lymphatic nodules, each nodule estimated to be from 1 to 4 cm3 in size, were treated by injection of N-CWS into the tumors. Complete regression of tumor was observed in seven out of ten tumors treated in five animals. Decrease of TAA-positive cells was also observed in PBL for all five treated animals. In one animal, the TAA-positive cells remained low for at least 280 days after treatment. This study documents that ADM treatment and intralesionally administered N-CWS are effective in the treatment of bovine leukosis. PMID- 2560857 TI - Natural mode of horizontal transmission of bovine leukemia virus (BLV): the potential role of tabanids (Tabanus spp.). AB - In order to evaluate the potential role of hematophagous insects in the natural spreading of bovine leukemia virus (BLV) infection in cattle, a 2-year survey was carried out involving sequential serological tests on 3328 cattle in three different areas. A parallel entomological study was run over the same period, using continuous trapping, in order to determine both the density and variations of horsefly (Tabanus spp.) populations in the close vicinity of the herds. After statistical analysis, this space-time study showed that: (1) There is a significant positive geographical correlation between the rate of incidence of BLV infection and the density of the horsefly population. (2) Seasonal variations in the incidence rate exist; the highest rates are generally observed during summer (from July of September), and the lowest during winter, spring and early summer (from November to mid-July). (3) There is a time link between the rate of sero-conversion and the variations in activity of the horsefly population. All these data combined would appear to indicate that tabanids play a considerable role in the spread of BLV under natural conditions. PMID- 2560858 TI - Cross-reactive antibodies to BLV and HTLV in bovine and human hosts with retrovirus infection. AB - Sera of 22 cattle naturally infected with bovine leukemia virus (BLV), of 2 calves vaccinated with BLV, and of 22 patients with human T cell leukemia virus type I (HTLV-I) infection were tested to BLV and HTLV-I by enzyme-linked immunosorbent assay (ELISA) and Western blotting (WB). Sera of 22 healthy cattle and from 22 healthy persons, and mouse monoclonal antibody to BLV-gp51, to HTLV-I p24, or to HTLV-I-p19 were also tested. Sera of virus-infected hosts gave significantly higher ELISA values than sera of healthy donors to both BLV and HTLV-I. The correlation between ELISA values of bovine sera to BLV and those to HTLV-I was r = 0.76, whereas that of human sera was r = 0.35. By WB and competitive WB assays, bovine sera that were ELISA-positive to BLV reacted with one or more of p12, p15, and p24 of BLV, and with only p24 of HTLV-I. Human sera that were ELISA-positive to HTLV-I reacted with p12 and p24 of BLV, and with one or more of p12, p15, p19, and p24 of HTLV-I. These results demonstrate that BLV and HTLV-I are capable of evoking cross-reactive immune response in at least some hosts under natural infection as well as by virus vaccination. PMID- 2560859 TI - Comparison of the agar gel immunodiffusion test and ELISA in the detection of bovine leukosis virus antibody in cattle persistently infected with bovine virus diarrhoea virus. AB - When six cattle persistently infected with bovine virus diarrhoea virus (BVDV) were inoculated with lymphocytes infected with bovine leukosis virus (BLV), a depressed antibody response to BLV was observed by ELISA which was due to a decrease in IgG1 synthesis. The ELISA was more sensitive and more reliable than the agar gel immunodiffusion (AGID) test in detecting BLV infection in cattle persistently infected with BVDV. Decreased antibody responses were manifested in the AGID test by negative, inconclusive or weakly positive reactions: only two of the six cattle developed antibodies that generated positive AGID reactions. PMID- 2560860 TI - Use of milk samples and monoclonal antibodies directed against BLV-p24 to identify cattle infected with bovine leukemia virus (BLV). AB - An indirect double-antibody sandwich (IDAS) enzyme-linked immunosorbent assay (ELISA) using milk samples was developed to identify cows infected with bovine leukemia virus (BLV). Two monoclonal antibodies (McAbs) were used. One, which was directed against BLV core protein p24, was used to coat ELISA plates; the other was used to prepare a horseradish peroxidase (HRP) conjugate directed against bovine immunoglobulin. The IDAS-ELISA detected antibodies directed against BLV p24 in 97% of the milk samples collected from known seropositive cows identified by the agar gel precipitation test (AGTP). Even when milk samples were diluted 1:50, 93% of the seropositive cows were identified. Only 0.43% of the 4000 milk samples collected in The Netherlands reacted nonspecifically. Nonspecific binding disappeared, however, when these samples were diluted 50 times in BLV-negative milk. In a comparative evaluation of BLV test-kits in various European laboratories, our IDAS-ELISA using McAb directed against p24 was one of the most sensitive. PMID- 2560861 TI - An international comparison of different laboratory tests for the diagnosis of bovine leukosis: suggestions for international standardization. AB - A collaborative study was conducted to compare the detection limit of different laboratory tests for antibodies against bovine leukemia virus (BLV). Serum and milk samples were tested in agar gel immunodiffusion (AGID), different modifications of indirect ELISA, blocking ELISA and ELISA procedures using monoclonal antibodies to BLV gp51 or BLV p24. The detection limit of reference serum E4 diluted 2-fold in negative serum gave a median value of 1:16 in AGID, indirect ELISA, and monoclonal ELISA p24, 1:128 in monoclonal ELISA gp51, and 1:1024 in blocking ELISA. The detection limit of a 4% immunoglobulin preparation of E4 diluted in negative milk showed median values of 1:800 in indirect ELISA, 1:1000 in monoclonal ELISA, and 1:2400 in blocking ELISA. None of the ELISA procedures could detect all the positive individual milk samples diluted 1:50. The AGID test is the official reference test for detection of antibodies against BLV. Reference serum E4 diluted 1:10 in negative serum must be scored positive in the AGID test. It is suggested that an international reference serum standard be established rather than an official recommendation of a particular ELISA test. PMID- 2560862 TI - The implication of BLV infection in the productivity, reproductive capacity and survival rate of a dairy cow. AB - The objective of the present study was to find out whether BLV infection, known to cause immunodisturbances in cows, might also bring about decreased productivity, reproductive rate and a shorter life span. More than 100 pairs of dairy cows, and a whole population of 3000 milch cows, were studied for this report. The findings revealed that a BLV-positive cow had a shorter life span than both its seronegative counterpart and the entire milch cow population. It also produced a total of 3.5% less milk and had a mean of 48 more days open than did the BLV-negative cow. The differences in survival rate were highly significant, while, at a level of 5%, those of productivity and reproductive rate were not. The implications of these findings are discussed. A highly significant correlation was also shown between BLV infection and the persistence of Trichophyton verrucosum infection in cows. The presented data indicate that BLV infection might affect the immune system of a cow to such a degree that it ceases to be productive enough to be kept within a herd. Thus it is usually culled before any severe symptoms of disease emerge. PMID- 2560863 TI - Phenotypic characterization of bovine lymphoblastoid cell lines. AB - Cytochemical and immunological markers were used to phenotype the bovine lymphoblastoid cell lines BL-3*, EBL-1, and EBL-2. Southern blot experiments were also performed to test these lines for the presence of proviral bovine leukemia virus (BLV). The BL-3* cell line, originally derived from a case of sporadic bovine leukosis (non BLV-associated) but later infected with BLV in vitro, was found to contain BLV provirus and expressed the BLV-encoded envelope glycoprotein BLV-gp51. BL-3* cells express surface IgM and cytoplasmic IgM as well as class II antigens, and greater than 95% were negative for the T-cell markers B26A, sheep erythrocyte (E) receptors and alpha-naphthyl butyrate esterase (alpha-NB). BL-3* thus appears to be B-cell derived. EBL-1 and EBL-2 were derived from cows with enzootic bovine lymphosarcoma; however, these cell lines were found not to be infected with BLV. Phenotypically, EBL-1 and EBL-2 are mature T-cells, as they were positive for the B26A epitope, alpha-NB, and E receptors. These cell lines also express class II major histocompatibility antigens, indicating an activated state. The T-cell phenotype of EBL-1 and EBL-2 raises interesting questions concerning the possible role of other retroviruses and non BLV-infected transformed T-cells in the development of EBL tumors. PMID- 2560864 TI - Porcine neutrophil function in the presence of virulent and avirulent Salmonella choleraesuis. AB - Porcine polymorphonuclear leukocytes (PMNLs) may be activated by bacteria to begin phagocytosis followed by oxidative and non-oxidative mechanisms of killing. The purpose of this study was to identify differences between virulent and avirulent Salmonella choleraesuis (S. choleraesuis) strains, 38 and 9 respectively, in their interactions with porcine PMNLs using five different assays. (1) Staphylococcus aureus (S. aureus) ingestion was determined by exposure of porcine PMNLs to a mixture of S. choleraesuis and 125I labeled S. aureus. There was a 2.98% and 22.20% decrease in S. aureus ingestion by mouse avirulent S. choleraesuis 9 and mouse-virulent S. choleraesuis 38 respectively. (2) Iodination of proteins was done by exposing zymosan stimulated porcine PMNLs to S. choleraesuis in the presence of 125I and measuring its incorporation into porcine PMNL proteins. This assay indicated a 73.7% and 74.7% decrease in iodination by S. choleraesuis 9 and S. choleraesuis 38, respectively. (3) Cytochrome c reduction was performed by using porcine PMNLs, zymosan, and S. choleraesuis to determine the bacterial effect on superoxide anion production. S. choleraesuis 9 and S. choleraesuis 38 inhibited superoxide anion production by 78.0% and 92.6%, respectively. (4) Lactoferrin release from porcine PMNLs was measured by an ELISA using the supernatant from the cytochrome c assay. Results indicate a 52.0% and 61.0% increase in lactoferrin release by S. choleraesuis 9 and 38 respectively. (5) The bactericidal assay was performed by counting cfus of S. choleraesuis after preliminary incubation with porcine PMNLs, followed by killing of extracellular S. choleraesuis and lysis of porcine PMNLs. Survival of S. choleraesuis 9 and E. coli (control) were 7.50% and 1.37%, respectively, in contrast to 52.62% survival of the virulent S. choleraesuis 38. These results indicate that both strains inhibited protein iodination and caused a slight increase in lactoferrin release, but the virulent S. choleraesuis 38 inhibited S. aureus ingestion, cytochrome c reduction, and survived porcine PMNL killing more effectively than the avirulent S. choleraesuis 9. PMID- 2560865 TI - [Balneotherapy at the early posthospital stage in the rehabilitative treatment of patients who have had an acute myocardial infarct]. AB - Sulfurated hydrogen baths under ECG and hemodynamic control were added to standard treatment of 99 patients who had suffered acute myocardial infarction 25 40 days before. Comparison with 62 matched controls showed a beneficial action of the baths on the clinical course of the disease, hemodynamics, coronary circulation, exercise tolerance. There was also a resultant reduction in the drugs doses. PMID- 2560866 TI - [Na,K-ATPase activity as one of criteria in identifying the risk group for development of late toxemia in pregnancy screening]. AB - Activity of erythrocyte Na+, K(+)-ATPase was studied in screening of pregnant women. Alterations in total antiradical status were estimated by means of estimation of erythrocytes resistance to peroxidation. 259 healthy women with pregnancy of 20-34 weeks, without any clinical manifestations of gestosis were examined. The enzymatic activity studied was decreased in erythrocytes of women both with nonimpaired and aggravated obstetric history; these groups of women were impaired with gestosis. Functions of Na+,K(+)-Atpase enzymatic system appear to be impaired during the preclinical steps of gestosis development. Alterations in activity of Na+,K(+)-ATPase in blood of pregnant women should be used in evaluation of risk groups for development of late toxicoses. PMID- 2560867 TI - [1,25-Dihydroxyvitamin D3 receptors in lymphocytes and T- and B-lymphocyte count in patients with glomerulonephritis]. AB - Content of receptors to hormonal form of vitamin D3, 1.25(OH)2D3, constituted 27.3 fmole/mg of protein in lymphocytes of peripheric blood of children with glomerulonephritis. In the patients concentration of total and ionized form of Ca2+ was decreased down to 2.04 mmole/L and 1.09 mmole/L, respectively, while an increase in parathormone (PTH) by 36% and a distinct decrease in 25(OH)D concentration (lower than 1.25 ng/ml) was found in blood; content of cAMP was also decreased in lymphocytes by 33%. At the same time, total content of T lymphocytes was decreased 1.5-fold in peripheric blood. Treatment with I hydroxyvitamin D3 (1-1.5 mg daily, within 4 weeks) led to normalization of total and ionized form of Ca2+ and of 25(OH)D, but did not affect the PTH content in blood. Concentration of the receptors to 1.25(OH)2D3 was elevated up to 39.7 fmole/mg after I week of the treatment, whereas it was decreased to the initial level 24.8 fmole/mg within 4 weeks; simultaneous alteration in the cAMP content was observed in lymphocytes. Treatment with 1-(OH)D3 normalized also the T lymphocytes content in peripheric blood. The data obtained suggest that under conditions of glomerulonephritis only high content of receptors to 1.25(OH)2D3 in lymphocytes enabled to perform the cell response to the hormone effect. PMID- 2560868 TI - [Resistance of heart valve bioprosthesis to the proteolytic effect of collagenase and elastase]. AB - Proteolytic destruction of pig aortal valves was studied at various steps of their preparation to implantation (native tissue, the tissue treated with terrylytine as well as with terrylytine and glutaric aldehyde) using pig pancreatic elastase and bacterial collagenase. The rate of the tissue destruction was estimated by means of monitoring an increase in content of protein and amino nitrogen in the hydrolysates. The tissue treated with terrylytine and glutaricaldehyde was 10-40-fold more resistant to proteolysis as compared with native heart valve tissue. Inadequate stabilizing effect of glutaric aldehyde on elastin, as compared with that on collagen, was found, when proteolysis of native and modified with glutaric aldehyde elastin from bovine cervical ligament and of calf skin collagen was studied using elastase and collagenase, respectively. PMID- 2560869 TI - [Functional changes in the synaptic membrane during in vitro interaction with bilirubin and serum albumin]. AB - A decrease in total activity of ATPase and in specific activity of Na+,K+-ATPase as well as less rapid and distinct decrease in specific activity of acetylcholinesterase were observed in synaptosomal membrane after binding of bilirubin in vitro. Irradiation with blue light of the bilirubin-containing membrane particles caused a more distinct decrease in activity of these enzymes. Blood serum albumin, added to the suspension of bilirubin-containing particles of synaptosomal membrane, affected significantly the alterations in activity of membrane-bound enzymes caused by the irradiation. Character of the effect depended on pH of the medium and presence of organophilic ligands in blood serum albumin molecule. PMID- 2560870 TI - [Primary culture of hepatocytes of newborn rats as a system for modeling anoxia and metabolic ischemia]. AB - A primary culture of hepatocytes from new born rats was used as an experimental model suitable for studies of the cell lysosomal apparatus and of molecular mechanisms involving cyclic nucleotides for initiation of lysosomal response to extremal conditions. Substrate deficiency and anoxia of the cultivated hepatocytes were found to cause distinct alterations in the state of lysosomal apparatus accompanied by an increase in free activity of acid phosphatase. PMID- 2560871 TI - [The effect of the synthetic analog of prostaglandin E2 sulprostone on the adenylate cyclase system in decidual tissue of women]. AB - Activities of adenylate cyclase and phosphodiesterase were studied in decidual tissue of women, pregnancy of which was interrupted within the first term by means of synthetic analogue of prostaglandin E2 sulprostone; decidual tissue obtained after surgical abortion served as a control. Content of cAMP in decidual tissue was decreased 4-fold after administration of sulprostone. At the same time, activity od adenylate cyclase was decreased 2-3-fold, while activity of phosphodiesterase was unaltered. These data suggest that the decrease in cAMP concentration occurred as a result of inhibition of its synthesis. After administration of sulprostone the adenylate cyclase activity was decreased 3- and 4.5-fold in response to specific stimulators NaF and forskolin, respectively, thus indicating that content and activity of G protein, and apparently of catalytic subunits, were decreased. Long-term effect of prostaglandins on decidual tissue appears to cause a heterologous desensitization of adenylate cyclase and this phenomenon is responsible for pharmacologic action of prostaglandins. PMID- 2560872 TI - [Metabolic status of selenium in pregnant women from a risk group of developing late toxemia]. AB - A decrease in activity of Na+, K(+)-ATPase, estimated in erythrocytes of women at the second half of pregnancy, served as a criterion in screening of the risk group tended to development of late toxicoses. In the risk group of women with pregnancy activity of glutathione peroxidase and content of selenium were studied in blood, simultaneously with estimation of osmotic resistance of erythrocytes. A decrease in activity of Na+, K(+)-ATPase was shown to be a reliable prognostic criterion in pregnant women with tendency to late toxicoses. At the same time, a decrease in glutathione peroxidase activity should be used as a distinct prognostic pattern only if content of selenium was as low as 95 pg/ml in blood of women with pregnancy. Alimentary and metabolic deficiency of selenium appears to be responsible for limitation of the glutathione peroxidase activity at the preclinical step of late toxicosis development. PMID- 2560873 TI - [Protein kinase activity and cAMP level in gastric mucosa in non-neoplastic diseases]. AB - Activity of cAMP-dependent and -independent protein kinases as well as content of cAMP were studied in biopsy preparations of gastric mucosal membranes obtained from patients with superficial gastritis, erosion, ulcer, adenomatous polyps and ulcer after treatment with laser irradiation. All the patterns studied were similar in preparations of adenomatous polyps and normal mucosal membrane. Activity of cAMP-independent protein kinases was increased under conditions of erosion, ulcer, gastroduodenal reflux and, especially, in ulcer tissue treated with laser. However, the ratio between cAMP-independent and -dependent phosphorylation was increased only in 50% of ulcerous diseases and in gastroduodenal reflux. The equilibrium of cAMP-dependent and -independent phosphorylation was not altered in the other impairments studied even though benign proliferation was stimulated using laser irradiation. PMID- 2560874 TI - [Study of the effect of rations containing dietary fiber on the level of total lipids and cholesterol in the blood and liver of the rat during a 6.5-month experiment]. AB - The influence of the following food fibers: wheat bran, citrus pectin, microcrystallized cellulose, methyl cellulose, was studied in experiments on male Wistar rats. Basing on the data obtained it has been suggested that the diets with pectin are conducive to secretion of general lipids and sterols from the liver. Comparison of the action of natural food fibers (wheat bran, pectin) with artificial fibers (microcrystallized cellulose, methyl cellulose) has not revealed any changes in the blood cholesterol level. At the same time a lower content of total lipids and cholesterol was recorded in the liver of rats that had received pectin and bran. PMID- 2560875 TI - [The infectious etiology of glomerulonephritis (a review of the literature)]. PMID- 2560876 TI - [Dynamics of the blood serum protein fractions in patients with insulinoma]. AB - Gel electrophoresis was used to study the blood serum of 13 patients with insulinomas who showed marked dysproteinemia in the preoperative period. Enucleation of the tumour or resection of the pancreas were performed depending on the location of the tumours. In the immediate postoperative period dysproteinemia became more pronounced, then a tendency to normalization of the protein spectrum occurred. Dysproteinemia was most pronounced at the zone of the starting peak where insulin-binding proteins are located. PMID- 2560877 TI - [Proteins and proteinase-resistant polypeptides of delta-endotoxins from Bacillus thuringiensis and their insecticidal activity]. AB - Proteins, solubility in alkaline buffer and proteinase-resistant polypeptides (PRP) of parasporal crystals (delta-endotoxin) from Bacillus thuringiensis were investigated. 19 sorts of crystals were divided into 7 types. Difference of crystalline proteins were coincident with their solubility and PRP. The characters of the protein, solubility and PRP were subspecies-specific basically, but in a few of crystals they were strain-specific. There was a closed relation between the biochemistry property of crystal protein and the toxicity in 2 species of insects. Crystal with protein of MW 130-138 kD kD or MW 130-138 kD and 60-65 kD, and PRP of MW 68-75 kD were high toxic in Bombyx mori (Lepidoptera), but most of them only showed weak or no toxicity in Culex fatigans (Diptera). Crystals containing more than 3 sorts of proteins with MW 15-138 kD in major protein and MW 35-75 kD in minor protein and PRP of 35-65 kD had very strong toxicity in Culex fatigans, but no toxicity in Bombyx mori. The construct of the crystal protein and the importance of the character in protein and PRP for the selection of expected strain and for the work of genetic engineering in Bacillus thuringiensis were discussed. PMID- 2560878 TI - [Preservation of cytomegalovirus by lyophilization]. AB - The preservation of cytomegalovirus (CMV) by lyophilization was first developed in China. In 4 to 10 days the typical CPE was up to + in the HEL cell line which was inoculated only 0.2 ml suspension of lyophilized strain AD169 or CMV isolated from milk breast. After preserved for 1 year the lyophilized cytomegalovirus strain still contained 1.5-2.0 lgTCID50/0.1 ml in titer of viral infection. The lyophilized strains have been used in the preparation of diagnostic antigen of cytomegalovirus in our department, also the strains were supplied to the whole country. PMID- 2560879 TI - [Herpetic encephalitis]. AB - On the basis of a literature review and own observations the author discusses the aetiology, pathogenesis, clinical course and treatment of encephalitis caused by herpes virus. Attention is called to the therapeutic methods. PMID- 2560880 TI - [Diagnosis of hemorrhage of Meckel's diverticulum using technetium scintigraphy]. AB - A 5-year-old boy is described with bleeding from Meckel's diverticulum diagnosed preoperatively by technetium scintigraphy. Despite limited indications to radioisotope methods in children they are valuable diagnostic procedures in preoperative diagnosis of the presence of ectopic tissue in Meckel's diverticulum. PMID- 2560881 TI - [Mechanisms of the conduction of cellular regulatory signals]. AB - A rat isolated heart was used to demonstrate a protective effect of phosphocreatine (10 mM) in total normothermic ischemia (25 min) with subsequent reperfusion with the Krebs-Henzeleit solution and in cardioplegia in the St Thomas Hospital solution. The effect consisted in reduction in ischemic and reperfusion contraction and a 2-3-fold increase in cardiac output as compared with the control. A 20 per cent increase in the calcium content in the solution did not influence the phosphocreatine effect. Application of tocopheryl phosphate (0.1 microM) had no effect on the ischemic contraction but reduced diastolic pressure on reperfusion and provided a better maintenance of the cardiac parameters. The tocopheryl phosphate action increased when used in combination with phosphocreatine. The findings show that phosphocreatine slows down the development of ischemic damage and onset of irreversibility while antioxidants exert a protective effect by preventing a reperfusion damage to the heart. PMID- 2560882 TI - [The effect of the ACTH(4-10) fragment on certain forms of adaptive behavior in mice of various genetic groups]. AB - CBA/Lac/Sto mice, C57/BL/6J mice and random bred mice with robertsonian translocation of chromosomes 8 and 17 (T1IEM mice) were compared under normal conditions and after ACTH4-10 injections. The rate of food procuring learning in U-shaped maze and in radial 5-arm maze was studied, and the ability of mice to extrapolate the direction of stimulus movement. Peptide i. p. injections (40 mkg/kg) stimulated the learning in U-shaped maze in all genotypes. T1IEM mice demonstrated better radial arm maze performance than CBA. Peptide injections tended to improve it in the former and to impair in the latter genotype. T1IEM mice demonstrated the ability for extrapolation, while CBA mice revealed no such ability. ACTH4-10 injections improved problem solving only in T1IEM mice. Cases when animals "refused" to participate in the experiment, were significantly rare in groups of all genotypes under peptide treatment. PMID- 2560883 TI - Age-related alteration in the catecholamine-sensitive adenylate cyclase system of the rat cerebral cortex. AB - Reduced adenylate cyclase (AC) activity of cerebral cortical membrane in the presence of guanine-nucleotide analog [Gpp(NH)p] was observed in 12-month-old rats (aged rats) compared with 2-month-old rats (controls). The EC50 value for Gpp(NH)p activation to AC was slightly but significantly increased in aged rats and the half time for maximal activation of AC by Gpp(NH)p, either with or without isoproterenol was also longer in aged rats. Digitonin-solubilized AC catalytic moiety had a lower activity in the aged rats than in controls. The 125I pindolol binding to beta-receptor in the cortical membrane from aged rats was decreased. GTP increased the Hill coefficient for isoproterenol displacement binding in controls, but such a change was not noted in aged rats. However, the percent increase of AC activity by 1 microM isoproterenol was greater in aged rats than that in controls. Furthermore, the EC50 value for isoproterenol stimulation of AC in control was lower compared with aged rats. Our results indicate that the decreased AC activity in aged rats may be due to a reduced receptor density, functional changes in GTP-binding protein (G protein) activity, and a reduced catalytic subunit activity. Compensatory changes in the beta receptor-G protein-AC coupling system may also occur during aging. PMID- 2560884 TI - [Pathophysiologic-nephrologic hypothesis of the protective function of an endogenous natriuretic and digoxin-like substance (endoxin) for residual nephrons]. AB - A dualistic function for the natriuretic ATP.inhibiting factor is formulated on the basis of a phenomenon like compensatory renal adaption of residual nephrons: 1. Support of the tubular sodium excretion to the maintenance of homoeostasis of the organism. 2. Inhibition of the forced up vasodilation of the afferent glomerular vessel. It should suggested that the renal vasoconstrictive effect is a part of a protective system for residual nephrons against the pathogenic hyperfiltration and hyperperfusion. PMID- 2560885 TI - Biochemical basis of platelet receptor function by GP IIb-IIIa and GP V. PMID- 2560886 TI - The role of complex formation and epidermal growth factor-like domains in the regulation of blood coagulation by the thrombomodulin-protein C system. AB - The explosive nature of the coagulation cascade led many scientists to investigate how it is regulated. Proteinase inhibitors such as antithrombin III inhibit active proteases of the coagulation cascade. Cofactors such as factor Va and factor VIIIa are proteolytically inactivated by activated protein C. Protein C is activated by the thrombin-thrombomodulin complex on the endothelial cell surface. Thus, the independent actions of the proteinase inhibitor system and the thrombomodulin-protein C system complement each other to maintain regulation of blood coagulation. The thrombin binding site of thrombomodulin was identified to be the fifth and sixth repeats of the epidermal growth factor-like domain. The same binding template contains sufficient information to block the functions of thrombin as a procoagulant. However, additional repeats are required for the activation of protein C. Thrombomodulin is the first example which illustrates that the epidermal growth factor-like domain functions as a binding template for thrombin and as a switch to turn off the procoagulant activity of thrombin as well as to trigger the protein C anticoagulant pathway. Epidermal growth factor like structures are found in many of the coagulation factors. Complex formation is a repeated theme not only in the blood coagulation cascade but also in the thrombomodulin-protein C anticoagulant pathway. PMID- 2560887 TI - Myeloperoxidase gene expression in acute leukemias. AB - Since myeloperoxidase (MPO) is considered to be a critical marker of differentiating acute myelogenous leukemia (AML) from acute lymphocytic leukemia (ALL), the analysis of MPO gene expression may provide further insight into the leukemia classification and the lineage fidelity of leukemia cells. By Northern blot hybridization using full-length MPO cDNA as a probe, approximately 66% of AML cells (3/4 M1 cases, 2/4 M2 cases, 15/15 M3 cases, 11/15 M4 cases, and 2/12 M5 cases) were found to express MPO mRNAs, whereas none of 18 ALL cases did. MPO mRNA was detectable when AML cells contained at least 10% peroxidase-positive cells. APL (M3) cells expressed high levels of mRNA in accordance with heavy staining for peroxidase. PMID- 2560888 TI - Acute and chronic effects of carbon monoxide on mitochondrial function. AB - The purpose of the present study was to examine whether there were some differences between metabolic responses induced by carbon monoxide (CO) poisoning and those by hypoxic hypoxia. Significant decrease in cytochrome oxidase (COX) activity in heart and liver was observed during acute exposure, while no change was observed in brain. When the control of respiration was investigated, cerebral energy homeostasis was maintained even after severe exposure. In order to document this phenomenon we measured blood flow in liver and brain, but increase in cerebral blood flow was not observed and the experiments failed to show a relationship between cerebral homeostasis and blood flow. The prolonged exposure to CO and hypoxia resulted in an increase in COX and these responses were obviously considered to be adaptive to chronic exposure. From the results of acute and chronic exposure it is considered to be difficult to determine if CO has the direct effect on the respiratory chain. Further studies of the mechanism of the CO toxicity may be necessary. PMID- 2560889 TI - [Characterization of the binding of [3H]-MK 801 to glutamatergic NMDA receptors in normal cerebral cortex and in Alzheimer's disease]. PMID- 2560890 TI - Molecular biology of human apolipoproteins B and E and associated diseases of lipoprotein metabolism. PMID- 2560891 TI - Occurrence of intracytoplasmic inclusion bodies in the digestive epithelium of fallow deer (Dama dama L). AB - Intracytoplasmic inclusion bodies were observed in the digestive epithelium of fallow deer (Dama dama L) suffering from bovine virus diarrhea/mucosal disease. Similar inclusion bodies were also found in the ruminal epithelium of fallow deer subjected to overfeeding by supplementary food. Inclusion bodies were not found in the upper alimentary mucosa of clinically healthy deer but were frequently found when these tissues were subjected to autolysis. At electron microscopical studies the inclusion bodies were found to consist of granular protein-like material encircled by a single membrane. Such inclusion bodies may constitute a non-specific degenerative cell response which could be elicited by diverse factors including autolysis. PMID- 2560892 TI - [Cytomegalovirus infection and pregnancy]. PMID- 2560893 TI - [Physiopathology of dilated cardiomyopathy]. PMID- 2560894 TI - Pyramidal cell topography of microtubule-associated proteins and their precipitation into paired helical filaments. PMID- 2560895 TI - Multiple excitatory amino acid receptor regulation of intracellular Ca2+. Implications for aging and Alzheimer's disease. PMID- 2560896 TI - The role of calcium in long-term potentiation. PMID- 2560897 TI - Links between long-term potentiation and neuropathology. An hypothesis involving calcium-activated proteases. PMID- 2560898 TI - Calcium- versus G-protein-activated phosphoinositide hydrolysis in synaptoneurosomes from young and old rats. PMID- 2560899 TI - Do activity-dependent changes in expression of regulatory proteins play a role in the progression of central nervous system neural degeneration? PMID- 2560900 TI - Calcium-activated neutral proteinases as regulators of cellular function. Implications for Alzheimer's disease pathogenesis. AB - Evidence is emerging that calcium-activated neutral proteinases (CANPs) not only participate in intracellular protein turnover but help to regulate the functional reorganization of cytoskeletal proteins in response to calcium and second messenger stimulation. The high concentration of CANPs in certain neurons has suggested prominent roles for this proteolytic system in neuronal and synaptic function. In addition to acting directly on specific constituents of the cytoplasmic and membrane-associated cytoskeletal networks, CANP may amplify its effects by modulating the activities of protein kinase C and possibly other kinases and phosphatases by limited proteolysis. Given its suspected involvement at the cytoskeleton-membrane interface, calcium-mediated proteolysis is an example of a metabolic process which, if impaired, could provide a unifying basis for the slow progressive development of diverse structural and functional abnormalities within neurons. The multiplicity of mechanisms regulating its activity makes the CANP system a vulnerable target for disruption from various sources. A working hypothesis is advanced that down-regulation (inhibition) of neuronal calcium-mediated proteolysis in Alzheimer's disease is one critical and early step in the development of neurofibrillary degeneration and altered membrane cytoskeleton dynamics, which leads to membrane injury, accumulation of abnormal proteins, and synaptic dysfunction. PMID- 2560901 TI - Regulation of membrane function through composition, structure, and dynamics. AB - Recent data reveal that membrane proteins can be regulated by the lipid environment of the membrane in which the proteins are located. Phospholipids and sterols are capable of regulating membrane protein activity. In some cases, the regulation is specific and may be crucial to cell function. The mechanisms by which this regulation is carried out have not been firmly established. However, the likely mechanisms include bilayer thickness, specific binding of lipids to sites on membrane proteins, and interactions between the surface of the protein and the surface of the lipid bilayer. PMID- 2560902 TI - Calcium homeostasis and aging: role in altered signal transduction. PMID- 2560903 TI - Cycling of Ca2+ across the plasma membrane as a mechanism for generating a Ca2+ signal for cell activation. PMID- 2560904 TI - Aging-related increases in voltage-sensitive, inactivating calcium currents in rat hippocampus. Implications for mechanisms of brain aging and Alzheimer's disease. PMID- 2560905 TI - [Low molecular weight heparin: an real alternative in thrombolysis in hemodialyzed patients. A trial of coronary thrombolysis]. AB - Fibrinolytic treatments pose serious problems in subjects at high risk for hemorrhage, such as those requiring chronic dialysis. A 36-year-old patient requiring dialysis for the last 14 years due to chronic kidney failure was hospitalized for unstable angina combined with calcified mitral stricture. Prompt coronary arteriography revealed recent intracoronary thrombi. The failure of drug treatment and the surgical indication in light of unstable angina led the authors to use low-molecular-weight heparin b.i.d. for 12 days with monitoring of laboratory parameters (anti-Xa activity, APTT). No thrombotic or hemorrhagic episode was recorded. Control angiography indicating partial lysis of the left and right intracoronary thrombi led to exact evaluation of the residual underlying stenoses. A double aortocoronary bypass was subsequently performed combined with replacement of the mitral valve. This case gives a glimpse of the potential value of low-molecular-weight heparin as a valid alternative to conventional fibrinolysis in subjects requiring dialysis. PMID- 2560906 TI - [Utilization of modern and traditional health systems in the rural Ivory Coast area]. AB - The authors carried out a questionnaire survey among the potential users of three rural health centers in Cote d'Ivoire. Between 58 and 90% of heads of family visit their zonal health center, between 18 and 50% also visit more distant centers. The utilization of other modern health structures is rare, but appeal to the herbalist healers is common and varies according to the characteristics of the families. Utilization of the two systems is not exclusive, 51% starting with the biomedical service. Of interrogated heads of family, 24% procure health care for others than members of their own family. PMID- 2560907 TI - [The antibiotic norseotricin as a feed additive]. PMID- 2560908 TI - [DNA restriction-modification systems]. PMID- 2560909 TI - [From alpha 2-adrenergic to endazoline receptors]. AB - Studies of the central hypotensive mode of action of the imidazolines of which clonidine is the leading molecule suggest the presence of non-catecholamine binding sites called imidazoline receptors. Our group showed that neither the endogenic ligand of alpha-adrenergic receptors, noradrenaline, nor any other tested catecholamine or phenylethylamine have hypotensive effects at the site of action of all imidazolines, the lateral reticular nucleus of the brainstem. In addition, a population of membrane binding sites which take up labelled clonidine and which are insensitive to noradrenaline have been demonstrated in the lateral reticular nucleus. An endogenic non-catecholamine substance whose structure is currently under identification and which is recognised by these receptors has been isolated from the brain tissues of various mammals. All this experimental evidence supports the hypothesis that the hypotensive effects of imidazoline-like substances are related to their action on brainstem receptors specific to this endogenic ligand which we propose to call endazoline. Rilmenidine, which has a chemical structure similar to that of the imidazolines, has a higher relative selectivity for the imidazoline binding sites than the reference molecule (clonidine). A central antihypertensive agent without the classical sedative effects associated with this class of drug could result. A study of the structure activity relationship is needed to confirm this hypothesis. PMID- 2560910 TI - [A case of Wilms' tumor and neuroblastoma of the kidneys in a child]. AB - Both kidneys in an 8-month-old boy contained tumours; Wilms' tumour was found in the right kidney and neuroblastoma in the left one. Nephroblastomatosis was observed in both kidneys. Multiple skin angiomas were observed after birth. Diagnosis of renal tumours was confirmed electron microscopically. PMID- 2560911 TI - Atrial natriuretic peptide: water and electrolyte homeostasis. AB - In the few years since its identification, a clear role for ANP in the regulation of water and electrolyte balance has emerged (Figure 3). The peptide is released in response to blood volume expansion, both acutely and gradually during changes in dietary sodium intake. Similarly, plasma levels are elevated in pathophysiological conditions such as cardiac and renal failure. It has become apparent that ANP has natriuretic, diuretic and vasorelaxant properties. Many of the original studies employed what we now know to be pharmacological doses of the peptide. However, recent reports have confirmed that small, sustained elevations in plasma ANP within or marginally above the 'normal' physiological range produce similar effects. A number of recent studies have tried to specifically address the physiological relevance of ANP. Although undoubtedly release by atrial distension and effective when infused to similar concentrations, atrial distension also has other effects via neural pathways. Thus, the demonstration that excretion of saline is impaired by atrial appendectomy (Benjamin et al, 1988) does not imply that this is only due to the absence of an atrial hormone. Goetz et al (1986) demonstrated that in the denervated heart, although ANP is still released, the excretion of a saline load is impaired. Similarly, in man, Richards et al (1988a) needed to infuse ANP to much higher plasma levels than those achieved by a saline load in order to reproduce the natriuresis. Although these experiments can be criticized, they confirm that ANP is not the sole mechanism for excreting a volume load, or for the natriuresis following atrial distension, but that these effects are likely to reflect the balance between ANP, AVP, the renin-angiotensin and autonomic nervous systems. In rats immunized against ANP (Greenwald et al, 1988), although the ability to excrete an acute saline load was impaired, long-term sodium balance was normal, suggesting that the rats were able to compensate for the absence of ANP. Many of the actions of ANP can be explained by antagonism of the renin-angiotensin-aldosterone system. Teleologically, it seems appropriate that a natriuretic hormone should counterbalance the major pressor and antinatriuretic hormones within the body. There is good evidence for cellular interactions between angiotensin, AVP, aldosterone and ANP at a number of discrete sites which are additional to the straightforward physiological antagonism of systems with opposing actions. ANP inhibits aldosterone secretion directly and may also reduce renal renin release. In the vascular tree there is evidence that ANP specifically blocks the vasoconstrictor actions of angiotensin II and possibly AVP.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2560912 TI - Importance of thermovision in radial mononeuropathy of compressive and ischaemic origin. AB - This prospective study was aimed to determine the diagnostic and prognostic values of Thermovision in compressive and ischaemic radial mononeuropathy. Thermography was carried out in case of 30 patients before and after treatment. Twenty healthy volunteers were included as control. Comparative analysis of the obtained particulars proved the diagnostic and prognostic values of Thermovision (t = 6.4, p less than 0.001) in radial neuropathy of compression-ischaemic origin. PMID- 2560913 TI - The canyon hypothesis. AB - The three-dimensional structure of human rhinovirus 14 has a deep surface depression or "canyon" encircling each of the twelve fivefold vertices. The canyon's surface is inaccessible to the broad antigen binding region of antibodies, permitting conservation of residues that might be required for host cell receptor recognition without danger of attack by the host's immune system. In contrast, the exposed surface features, where neutralizing antibodies are known to bind, change rapidly under pressure from the host's immune system. It was, therefore, hypothesized that this depression was the site of receptor attachment. Similar, but smaller, depressions had been observed previously on both the hemagglutinin and neuraminidase spikes of influenza virus. These have also been shown to be the site of host cell interaction. Although support for the canyon hypothesis was only circumstantial in the first place, there are now extensive confirmatory data. These include site-specific mutations of residues in the canyon and conformational changes induced in the canyon by the binding of small organic molecules, all of which alter receptor attachment. The strategy used in human rhinovirus 14 to protect the viral receptor attachment site from immune surveillance may be utilized not only in other picornaviruses but also in many other types of viruses including human immunodeficiency virus. PMID- 2560914 TI - Immunological surveillance in a pseudorabies quarantined herd using gilts and their progeny as sentinels. AB - Specific pathogen free gilts and their progeny were evaluated to use as sentinels in a pseudorabies virus (PRV) infected herd by immunologically monitoring for PRV seroconversions. Time intervals targeted were pre- and post-PRV vaccinations, herd exposure, and farrowing to finishing. Post-PRV vaccinations, gilts showed low PRV lymphocyte stimulation and humoral responses. Following herd exposure, control gilts PRV seroconverted and PRV vaccinated gilts increased (2 to 4 times) in virus neutralization (VN) titers. Sixty-seven percent (4/6) of the progeny from a control gilt were PRV seropositive at finishing. Progeny from PRV vaccinated gilts were depleted of passive immunity by week 7, and were seronegative until week 9. At finishing 47% (14/30) of them were PRV seropositive indicating exposure to PRV. The VN test was not sensitive enough to detect weak positive serums, noted as positives by latex agglutination (LA) test, ELISA, and Western blots. The gilts and progeny detected PRV, respectively, in the herd housing quarters and in the farrow to finish facilities. A strategy for future sentinel experimental surveillances using primarily the LA test is proposed. PMID- 2560915 TI - In vitro neutralization of antigenic variants of bluetongue virus is related to in vivo protection. AB - Neutralization resistant variants of bluetongue virus serotype 10 were selected with a neutralizing monoclonal antibody. Three variants were selected for further characterization. One of these variants was completely resistant to neutralization, while the other two variants showed intermediate resistance to neutralization as compared to the parent virus. The completely resistant variant failed to bind the selecting monoclonal antibody as determined by immune precipitation and enzyme linked immunosorbent assay; whereas, the other two variants bound antibody to a similar extent as the parent virus as determined by these tests. The ability of the variants to bind monoclonal antibody correlated with passive protection in the newborn mouse model. These results indicate that the ability of the virus to bind antibody is directly related to in vivo protection and that in vitro neutralization and in vivo protection are also related. PMID- 2560916 TI - Sex-dependent, early cytokine production by NK-like spleen cells following infection with the D variant of encephalomyocarditis virus (EMCV-D). AB - Spleen cell cultures from diabetes-resistant ICR Swiss females exhibited an increase in expression of Ia antigens 24 hours post-infection (PI) with EMCV-D while comparable spleen cell cultures from diabetes-susceptible males of this strain did not exhibit this increase in Ia antigens expression. A monoclonal antibody specific for mouse interferon-gamma (IFN gamma) eliminated this increase in Ia antigens expression. Interferon-gamma (IFN gamma) and interleukin 2 (IL-2) production by EMCV-D-infected spleen cell cultures were monitored at 4-hour intervals for 24 hours. Female spleen cells produced IFN gamma earlier (less than 16 hours PI) and in greater amounts than did comparably treated male spleen cells. Addition of a monoclonal rat anti-mouse IL-2 to virus-infected cultures did not significantly affect the early (less than 16 hours PI) production of IFN gamma by spleen cells of females. Treatment of the spleen cell donors with rabbit anti-asialo GM1 (AAGM1) abolished early production of IFN gamma in virus-infected female spleen cell cultures and reduced the early IL-2 production by infected male and female cells. These results suggest that an NK-like cell is responsible for the early female IFN gamma production; this may be a factor in the resistance of female ICR Swiss mice to EMCV-D-induced diabetes. PMID- 2560917 TI - Mouse thymic virus-mediated immunosuppression: association with decreased helper T cells and increased suppressor T cells. AB - Mouse thymic virus (MTV) is a herpesvirus which, when administered to newborn mice, induces an extensive but temporary thymic necrosis associated with immunosuppression. In the present study, the T cell subsets in the thymus of MTV infected newborn C57Bl/6 mice were evaluated at 4, 7, 14, 28, 56, and 84 days after infection, using labeled monoclonal anti-CD4 and anti-CD8 antibodies with two-color flow cytometry. At 7 and 14 days, the percentages of CD4+8- and CD4+8+ cells were significantly decreased whereas the percentage of CD4-8+ cell was increased. At days 28 and 56 percentages had returned to normal. These results indicate that the virus has an affinity for CD4+ T cells (helper cells and their precursors). Increased percentage of CD4-8+ T cells (suppressor cells) is also associated with depressed immune functions in MTV infected newborn mice. PMID- 2560918 TI - Comparisons of the structural proteins of avian infectious bronchitis virus as determined by western blot analysis. AB - The antigenic diversity of ten strains of avian infectious bronchitis virus (IBV) was examined by Western blot analyses using polyclonal antisera specific for the Massachusetts 41 (M41), Gray, Arkansas DPI (Ark DPI), Connecticut (Conn) and Australian T (Aust T) serotypes. Although antigenic variation was found in all three structural viral proteins, the matrix protein appeared to be antigenically the most highly variable. Four distinct antigenic groups, which did not correspond to virulence or pathotype, could be defined according to the variations observed in the matrix protein. Somewhat less variation was seen in the spike polypeptide. The only variation in the nucleocapsid protein was indicated by the lack of a detectable reaction between the Aust T antiserum and the Ark DPI nucleocapsid protein. Antisera made against M41 had the broadest reactivity while antisera against Aust T, the only strain tested which was exotic to the U.S.A., had the greatest specificity. PMID- 2560919 TI - [Signal transmission using second messengers]. AB - Intercellular communication needs signal molecules (hormones, neurotransmitters, growth factors) emitted by a cell and recognized by targets. A widespread mechanism of cellular action of these molecules involves a membrane receptor, a GTP-dependent coupling protein, an effector which modulates the intracellular concentration of a second messenger (cAMP, cGMP, inositol triphosphate, Ca++), and a target for final effect of this second messenger (kinases, channel). Hormonal systems and visual processes in vertebrates rod outer segments are the best known examples of this mechanism; they are taken here to illustrate the state of knowledge of the interactions between the actors of the transductional enzymatic cascades. PMID- 2560921 TI - Hydroxylapatite coatings bond chemically to bone. PMID- 2560922 TI - [Erythrocyte cation transport in arterial hypertension: interrelation with the renin-angiotensin-aldosterone system and the atrial natriuretic factor]. AB - Red cell membrane Na(+)-K+ transport systems, renin-angiotensin-aldosterone system (RAAS) and atrial natriuretic factor (ANF) were studied in a group of 50 mild essential hypertensive patients (n = 25 for each group) age, sex and blood pressure matched. Na(+)-K+ ATPase and intracellular Na+ (Na+ i) were significantly different between the two groups (p less than 0.01). A slight difference was also seen for the Na(+)-K+ cotransport (p less than 0.05) as a likely consequence of the differences in the methodology of Na+ charge to study its efflux from the red cells in vitro. A negative correlation (r = -0.47, p less than 0.01) was observed between ANF and Na(+)-K+ cotransport suggesting an interrelationship of the two systems in the homeostasis in body fluid and electrolytes. PMID- 2560920 TI - Steroid and barbiturate modulation of the GABAa receptor. Possible mechanisms. AB - This review describes the modulation of the GABAa receptor by steroid hormones and barbiturates and proposes guidelines for further research. Having examined the complex organization of the GABAa receptor complex and the multiple allosteric interactions between its drug and transmitter/modulator binding sites, the possibility that conformational changes of the receptor molecule may explain most of its characteristics is explored. On the basis of considerable evidence, we propose that the GABAa receptor may adopt as many as five different conformations. However, the heterogeneity of central GABAa receptor binding cannot only be explained by different configurations of a single protein. It also has been shown that different GABAa receptor subtypes exist within different brain regions. These receptor subtypes may differ from each other in their subunit composition. By describing the GABAa receptor as a macromolecular complex that may adopt different conformations and whose subunit composition may vary, it becomes possible to understand the molecular mechanisms by which steroid hormones modulate the receptor. This has led to two models of hormone actions. A first model addresses the direct effects that steroids exert on the GABAa receptor and predicts that steroid hormones may cause the conformation of the receptor complex to change between active and inactive states. A second model, which addresses the observed heterogeneity of GABAa receptor binding within the brain, suggests that steroid hormones may change the expression of the different subunits of the receptor complex by acting at the genomic level. This review complements other recent reviews describing the modulation of the GABAa receptor (Olsen and Venter, 1986; Gee, 1988). PMID- 2560923 TI - [Endogenous digoxin-like factor: serum concentration and interrelations with the electrolyte picture in normal subjects]. AB - Endogenous Digitalis-Like Factor (DLF) is a putative hypothalamic Na+,K+-ATPase inhibitor that mediates natriuresis in response to intravascular volume expansion or sodium loading. The precise structure of this substance remains unknown; however, it cross-reacts with antibody to digoxin. Using a radioimmunoassay, we measured DLF concentrations in 26 normal subjects: mean value of this factor was 0.512 ng digoxin-equivalents/ml +/- 0.038 SEM; DLF correlated significantly with serum sodium levels (r = 0.59 - p less than 0.01) and daily urinary sodium excretion (r = 0.48 - p less than 0.05). Our results confirm that endogenous digitalis-like factor has a physiological role as regulator of natriuresis, in response to plasma sodium concentrations. PMID- 2560924 TI - [Metabolism of NAD+ in the nuclei of human placenta]. AB - It has long been known that the major function of NAD+ is as an electron carrier in various biological oxidation-reduction systems. From many papers it is evident that NAD+ is involved as substrate in ADP-ribosylation reactions. We have focused our attention on two chromatin enzymes: NMN-adenylyltransferase that catalyzes reversible synthesis of NAD+ utilizing ATP and NMN, and poly(ADP ribose)polymerase that covalently modifies nucleosomal proteins through poly ADP ribosylation reactions. Here we provided evidence of these activities in a system of isolated nuclei from human placenta. The data presented in this report show that purified nuclei might be useful to study the nuclear location of these enzymes and their reciprocal interactions. PMID- 2560925 TI - [Actions of retinol and retinoic acid on hepatoma H4 cultures]. AB - The amount of vital cells recovered, their morphology (studied by SEM) and some of their biochemical aspects concerning the differentiation processes (aerobic glycolysis, cell production of cAMP and CEA) were investigated in a strain of H4 hepatoma cultured for 8 days in the presence of 5 microM retinol (R) or retinoic Acid (RA). Vital cell recovery is slightly reduced either by R or RA treatment. Flattening of the cell shape and reduction of the plasma membrane prolongations and of intercellular bonds are observed in the R-treated cells but to a greater extent in those treated with RA. Aerobic glycolysis is decreased in the R-treated cells but increased in those treated with RA. Such events could be related to the regressive processes observed in the RA-treated cells. cAMP cell content is increased to a greater extent in the R-treated cells than in those treated with RA. CEA cell content is greatly decreased in the RA-treated cells but only slightly in those treated with R. Therefore, the treatment of HA hepatoma cells with 5 microM R or RA, while reducing cell growth only slightly, does not cause evident and unequivocal morphological and biochemical events related to cell redifferentiation. PMID- 2560926 TI - [Purine metabolism: determination of PRPP-amidotransferase and PRPP-synthetase in lymphocytes from patients with chronic lymphatic leukemia (CLL)]. AB - Phosphoribosyl-1-pyrophosphate (PRPP) amidotransferase is the "key anabolic enzyme" of purine nucleotide synthesis; PRPP synthetase connects the pentose cycle with the same pathway. We have studied their behavior in 5 control subjects and in 8 affected by CLL. Determination of PRPP amidotransferase was carried out through the evaluation of 14C-glutamic acid (released by 14C-glutamine) in the incubation mixture. PRPP synthetase was followed by adding ATP and ribose 5 phosphate to the incubation mixtures, and by evaluating the PRPP formed through the release of CO2 in a coupled reaction. In the case of PRPP-amidotransferase, our values are in the range reported in the literature: in patients affected by CLL, the enzyme activity is much higher and the increase is more evident when values referred to the patients, than when to the cells. Our values of PRPP synthetase are consistent with those of Peters and Veerkamp, but no definite conclusion is possible in the case of leukemic patients. PMID- 2560927 TI - A rapid method of Southern blot analysis using polyacrylamide gel electrophoresis and vacuum blotting transfer techniques. PMID- 2560928 TI - A new method for storing animal tissue prior to mtDNA extraction. PMID- 2560929 TI - Performing nucleic acid reactions using predispensed lyophilized reaction mixtures. AB - A system is described in which manipulations of nucleic acids are performed in wells containing predispensed lyophilized reaction mixtures requiring addition of only nucleic acid. This allows increased reproducibility for single-step reactions (e.g., restrictions and ligations), as well as improved productivity for complex reactions (e.g., sequencing). Enzymes, co-factors, nucleotides and buffers can be dried and stored at room temperature without loss of essential function. When used for DNA sequencing, hundreds of templates a day can be sequenced with the potential to determine megabase amounts of sequence per week. PMID- 2560930 TI - A rapid visual method for the identification of 4- or 6-base restriction endonuclease sites. PMID- 2560931 TI - Genetic approaches to cytoskeleton function and the control of cell motility. PMID- 2560932 TI - Monoclonal antibodies that specifically inhibit GM-CSF- and IL-3-dependent growth of human monocytic leukemia cells. AB - We describe monoclonal antibodies (mAbs: anti-MaG-1, TGI-1, TGI-5, and TGI-6) that block the proliferation of AML-193 cells in response to GM-CSF or IL-3 and do not affect the proliferation of AML-193 cells in response to G-CSF and IL-2 driven proliferation of Kit 225 cells. However, none of the mAbs tested had any stimulative effect on the proliferation of AML-193 cells. The mAbs (anti-MaG-1, TGI-1, -5, and -6) could inhibit the binding of [125I]GM-CSF to AML-193 cells. We were able to purify MaG-1 Ag by anti-MaG-1 affinity chromatography. Thus, the MaG 1 Ag and the Ags recognised by mAbs (TGI-1, -5, and -6) may be associated with the receptor for GM-CSF or IL-3 or a structure close to the receptor for GM-CSF or IL-3. PMID- 2560933 TI - Expression of three recombinant homodimeric isoforms of PDGF in Saccharomyces cerevisiae: evidence for difference in receptor binding and functional activities. AB - Three recombinant homodimeric isoforms of platelet-derived growth factor (PDGF) were produced and purified in milligram quantities by expression of PDGF A- and B chains in yeast cells. Structural analysis of the purified short and long variants of PDGF-AA (PDGF-AAS and PDGF-AAL) and PDGF-BB showed that they had been properly processed and assembled into dimers. PDGF-AAS and PDGF-AAL were found to bind only to the PDGF A-type receptor on human fibroblasts, with affinities of 0.1 and 0.2 nM, respectively. PDGF-BB bound to cells with A- and B-type receptors and to cells with B-type receptor only with affinities of 0.6 nM in both cases. Each fibroblast appeared to express about 4-5 times more B-type receptors than A type receptors. The maximal mitogenic response to PDGF-BB of human fibroblasts was almost 2-fold higher than that induced by either of the two PDGF-AA forms. The three isoforms of PDGF also stimulated growth in soft agar of human fibroblasts with PDGF-BB inducing a higher maximal response. PMID- 2560934 TI - On the origin of antibodies to immunoglobulin genetic markers in rheumatoid arthritis. An outline of a novel concept of the nature of rheumatoid arthritis- transduction in man. AB - Anti-Gm's in rheumatoid arthritis patients detect products of the Mendelian genes G1ma, G1mx, G3mb, G3mc, G3mg, G3mst. Commonly, these anti-Gm's are specific for other individuals immunoglobulin allotypes. Reasons are given for the contention that such anti-Gm's in patients with rheumatoid arthritis are indicative of the expression of nonnominal allotypes, the genes for which have been transferred from one individual to another by means of a B cell virus. This process - akin to transduction and occurring after the tolerance induction period - may lead to a chimaeric state of the B cell compartment in rheumatoid arthritis patients. Special features of Ig gene regulation in normal B cell progression are allelic exclusion and the excision-expulsion-rejoining of DNA segments. Perturbation of B cell regulation by external genomes may be favoured by allelic exclusion. The DNA excision-rejoining processes may have consequences for concomitant (viral) nucleic acid assembly, particularly for viruses sharing nucleotide sequences with the Ig switch regions. PMID- 2560935 TI - 5-Lipoxygenase inhibition by antifungal azole derivatives: new tools for immunosuppression? AB - The effect of novel azole derivatives (itraconazole and fluconazole) on 5 lipoxygenase activity was examined using human polymorphonuclear leukocytes (PMNL) as the enzyme source. The novel imidazole derivative itraconazole proved to be a potent inhibitor (IC50 2 X 10(-6) M) of 5-lipoxygenase activity. In contrast, fluconazole, an antifungal agent with bistriazole structure, had no effect on the Ca2+ ionophore A 23187-induced formation of 5-lipoxygenase metabolites in PMNL. PMID- 2560936 TI - Preparation of leukotriene B4 antibodies in sheep. AB - The antibody response of eight sheep immunised against leukotriene B4-keyhold limpet haemocyanin (LTB4-KLH) was investigated. Four Suffolk x Mule sheep and two Soay sheep received subcutaneous injections of conjugate over a period of 11-21 weeks, and two Suffolk x Mule sheep received intramuscular and subcutaneous injections for 11 weeks. All eight animals reacted successfully to immunisation and produced antibodies of high affinity and specificity for LTB4. The antiplasma could be used for radioimmunoassay at dilutions of 1/10(5)-1/10(6) and the highest titres were obtained in Soay sheep. Intramuscular immunisation produced a faster but less sustained response than subcutaneous injections. The uniformity of response between animals in relation to the immunisation regime and the physico-chemical similarity of the antibodies suggest that the quality of the conjugate is the most important factor for successful antibody production. PMID- 2560937 TI - [Clinical and neurophysiologic tests in the normal elderly]. AB - Clinical and neurophysiological examination was performed in 35 normal aged subjects (72 +/- 6.4; range 65-86 years). Seventeen showed abnormal ankle reflex (48.6%), one case had patellar areflexia as well (2.9%), six had malleolar appalesthesia (17%), and five revealed both areflexia and apallesthesia (14%). Electromyography and motor conduction were normal in every case. Sural nerve conduction velocity and the amplitude of the sensory evoked potential showed no differences between subjects with and without clinical abnormalities. It is concluded that a) reflex and vibration sensibility changes in the elderly are not consequence of peripheral nerve involvement and b) there is no reason to believe there is an "aging neuropathy". PMID- 2560938 TI - Virus induced adherence of monocytes to endothelial cells. AB - In this study we have demonstrated that infection of human umbilical vein endothelial cells (HUVEC) with Herpes simplex virus type 1 (HSV-1) resulted in an increased adherence of monocytes (MC). This enhanced adherence occurred at 3 h post infection (p.i.) when about 20% of the monolayer is infected and when there is no cytopathic effect observable in the monolayer. The adherence of human MC to virus-infected HUVEC monolayers proved to be effective and reproducible if a multiplicity of infection (MOI) of ten and a ratio of number of MC to number of HUVEC of 5 was used. The increased adherence was also induced by incubating non infected HUVEC with the 'supernatant medium' of the HSV-1 infected cells, showing that soluble factors induced by viral infection are responsible for the increased adherence. The augmentation of MC adherence to infected endothelium was sensitive to tunicamycin treatment, suggesting that the MC adherence is probably mediated by glycoproteins expressed on the HUVEC membranes by virus infection. PMID- 2560939 TI - Recent advances in the genetics of the clostridia. AB - Several laboratories around the world have started work on genetic analysis of clostridia. Interest in this diverse group of anaerobic organisms has grown with increasing awareness of the benefits that may accrue from their biotechnological exploitation. Research to date has focussed on construction of shuttle vectors containing replicons from clostridial and streptococcal plasmids, development of methods for transferring genes, and molecular cloning of genes--especially those involved in toxigenicity, fermentative metabolism and polysaccharide utilization. In selected species gene transfer by protoplast transformation, electroporation and conjugation has been accomplished and transposable elements have been introduced. It can be anticipated that our understanding of the molecular biology of these interesting organisms will grow rapidly in the future, bringing with it improved prospects for rational biotechnological exploitation. PMID- 2560940 TI - Rotavirus identification in jejunal juice and stools of acute and chronic forms of infantile gastroenteritis. AB - 1. The present study investigates, by means of a combined enzyme immunoassay for rotaviruses and adenoviruses (EIARA), the occurrence of rotaviruses in stools and jejunal juices from 31 children with acute diarrhea and 18 with chronic diarrhea. 2. Stools from 8 acute cases contained rotaviruses (26%). In two of these cases rotaviruses were also detected in the jejunal juice. 3. In the chronic diarrhea group we identified rotaviruses in the stools of one patient and in the jejunal juice of another. 4. Some of the electropherotypes of the rotaviruses identified showed different patterns of RNA migration. 5. Abnormalities of the jejunal mucosa were characterized in 6 acute rotavirus-positive cases. No morphological or functional abnormalities of the intestinal mucosa were observed in the chronic diarrhea rotavirus-positive cases. PMID- 2560941 TI - Brachial plexus neuropathy in a gymnast associated with chicken pox. AB - A case report is presented of brachial plexus neuropathy in an international gymnast in association with chicken-pox and the clinical features, investigations and prognosis are discussed. PMID- 2560942 TI - The negative aspects of granulocyte transfusions. PMID- 2560943 TI - [Detection of the HBX gene product-HBxAG in primary hepatocellular carcinoma, tumor-adjacent tissues and serum in 7 cases]. PMID- 2560944 TI - [Effect of EB virus on the pathogenesis of primary Sjogren's syndrome]. PMID- 2560945 TI - [Trace elements in the serum and cancerous tissue in patients with trophoblastic carcinoma]. AB - In the serum and cancer tissue of 155 patients with trophoblastic tumor and in 114 patients with benign or malignant gynecologic tumor, 22 elements (17 trace and 5 major elements) were determined by neutron activation analysis. The serum and tissue level of Cr, the serum Zn level in patients with hydatidiform mole was significantly higher than that of the control group (P less than 0.01). The serum Se level in Patients with invasive mole and choriocarcinoma was lower (less than 1.0 mumol/l), but Cu/Zn ratio was higher (greater than 2.0) in comparison with that of the controls (P less than 0.01). With remission of the disease levels of the essential trace elements went up approaching the normal levels. There was a negative relationship between the serum Zn level and hCG content. The authors point out that the low level of Zn and Se, the high level of Cr may be related to the development of trophoblastic tumors. PMID- 2560946 TI - [The so-called sclerosing hemangioma of the lungs]. AB - A 12-year old girl had a focal density in the lung found by checking CT. Structural features were those of the so called sclerosing hemangioma. Analysis of structure in various methods and of references supported authors' interpretation that lesion represented a late consequence of pecul a obliterative mainly venous process with prevailing plasmacytic feature. Characteristic segmental destructive venous lesion conused segmental obliterat on and hyalinization. Similarity of the lesion to intravascular fasciitis was discussed. Peripheral accumulation of pneumocytes around the lesion seemed to be secondary. To name it pneumocytoma would be unproper and a possibility of vascular tumorous character could not based on respectable analogies. A closer etiopathogenetic specification of the so called sclerosing hemangioma of the lung remains to be accomplished. PMID- 2560947 TI - [ATPase activity, endogenous depression, lithotherapy and electroconvulsive therapy]. AB - The author gives a brief account of the action of ATP-ases in the organism. He describes the effect of different substances on the action of these enzymes, whereby special attention is paid to the action of psychopharmaceutical preparations, incl. lithium salts, on the activity of Na+K(+)-ATP-ase. In the experimental part he investigates the activity of ATP-ases in endogenous depressions in our population, the influence of Li+ and electroconvulsions on these enzymes. The results are consistent with data in the literature. Changes in the ATP-ase activities in the course of treatment imply that these enzymes are not genetic markers of depressions. The different effect of therapeutic preparations on the activity of ATP-ases indicates that influencing ATP-ase activity in relation to depressive conditions cannot be considered causal. PMID- 2560948 TI - Activation by cathepsin G of latent gelatinase secreted from rat polymorphonuclear leukocytes. AB - Rat polymorphonuclear leukocytes secrete a latent gelatinase with a molecular weight of about 96 kilodaltons (kDa). Activation of the latent 96-kDa gelatinase by cathepsin G was studied by using sodium dodecyl sulfate-substrate polyacrylamide gel electrophoresis. Cathepsin G activated the 96-kDa gelatinase by converting it to two lower molecular-weight species of 76 and 61 kDa, which were slightly different from the gelatinase species generated by treatment with 4 aminophenylmercuric acetate, an activator of latent gelatinase. PMID- 2560949 TI - Inhibition of adenosine 3',5'-cyclic monophosphate phosphodiesterase by flavonoids. III. AB - Sixty-one flavanones, twenty-six isoflavones and eight other flavonoids, obtained from Sophora tomentosa, S. flavescens, Scutellaria baicalensis and other medicinal plants or synthesized, were tested for their inhibitory activity against adenosine 3',5'-cyclic monophosphate (cAMP) phosphodiesterase from beef heart. The structure-activity relationships were investigated. PMID- 2560950 TI - [Memory, learning and glutamate receptor]. AB - After a brief tetanic stimulation of an excitatory pathway in the hippocampus, synaptic transmission through the tetanized pathway is facilitated for a long period. This phenomenon is called long-term potentiation (LTP), and has been regarded as a neuronal correlate with learning and memory. Features of LPT in Schaffer collateral-CA1 neuron are calcium-dependency, input-specificity, cooperativity and sensitivity to blockers of N-methyl-D-aspartate (NMDA) receptors. These features can be explained by properties of NMDA receptors. LTP in mossy fiber-CA3 neuron has features different from those in field CA1: It is insensitive to blockers of NMDA receptors and does not show cooperativity. Quantal analysis of transmission through mossy fiber synapses before and after generation of LTP reveals that LTP resulted from facilitated release of transmitter. Participation of the metabotropic glutamate receptor is suggested. PMID- 2560951 TI - [Reproducibility of localized protein spectra with a 1.5 T whole body nuclear magnetic resonance scanner]. AB - The CODEX method was employed for volume selective proton NMR spectroscopy with a whole body imager in order to control the quality of reproduction with phantoms. The spectral resolution and time stability during typical measuring times are almost better than 3.10(-8) and are usually sufficient for the requirements of high resolution in vivo proton spectroscopy. PMID- 2560952 TI - Interaction of anti-ulcer drugs with the gastric proton pump. AB - In this review consideration is given to anti-ulcer drugs interaction with the gastric H(+)-K+ ATPase. The review has been divided into three sections. First, properties of the gastric proton pump are described in terms of structure, biological activity and ions transport activity, followed by an account of interactions involving antisecretory agents. Emphasis is given to a new class of drugs (substituted benzimidazole) that shows a unique antisecretory action and is safe and effective for short-term treatment of patients with duodenal or gastric ulcers. The final section briefly examines future directions for the production of more selective inhibitors of the gastric proton pump. PMID- 2560953 TI - Prevention of myocardial reperfusion injury with free radical scavengers. An experimental study. AB - The changes in endogenous superoxide dismutase (ESOD) during myocardial ischemia and reperfusion and the efficacy of oxygen free radical scavengers in myocardial protection were investigated in an isolated heart model connected with the recirculating nonpulsatile perfusion circuit. Subjected to a 2-hour period of global ischemia (27 C), the heart was reperfused with 37 C oxygen diluted auto blood for 60 minutes. Superoxide dismutase plus catalase was added into the cardioplegic solution and reperfusates. ESOD activity was measured by pyrogallol method. The results of the experiment showed that ESOD activity after ischemia and reperfusion was decreased and the addition of oxygen free radical scavengers (SOD and CAT) to the cardioplegic solution and the reperfusates greatly reduced the leakage of myocardial enzymes, coronary vascular resistance, and the ultrastructural damages of the myocardium. These results suggest that the use of SOD and CAT may inhibit myocardial reperfusion injury by scavenging oxygen derived free radicals. PMID- 2560954 TI - Small hepatocellular carcinoma. DNA content and biological characteristics. AB - The DNA content of 12 small hepatocellular carcinomas (SHCC, less than 3 cm in diameter) and 26 large hepatocellular carcinomas (LHCC) was quantitatively determined by means of TV-image analysis. The results showed that 8 patients (66.7%) with SHCC had DNA stem lines in diploid (2C), and 24 (92.3%) with LHCC had DNA stem lines in aneuploid (AN) (P less than 0.01). The incidences of tumor capsule breaking and cancerous thrombosis were 16% and 20% respectively in SHCC, significantly lower than 84% and 80% in LHCC (P less than 0.01). The 5-year survival rate was 75% in patients with SHCC after operation, much higher than 46.2% in patients with LHCC. No relations were found between DNA content, tumor size, pathological grading and serum AFP values of HCC. The results suggest that SHCC less than 3 cm in diameter reflects the early changes of biological characteristics; HCC of 3 cm in size may be at an important period when the changes of DNA stem lines and biological characteristics would occur; SHCC is possible to be determined by serum AFP values; and that the survival rate of the patients can be further improved by early finding and treatment of SHCC less than 3 cm in diameter. PMID- 2560955 TI - Effects of FDP and Danshen on renal cortical Na-K-ATPase activity in rats after treatment with renal ischemia and gentamicin. AB - The main purpose of our study is to investigate the possible protective effects of Fructose 1-6 diphosphate (FDP) and Danshen (Salvia Miltiozzhiza Bunze) on renal cortical Na-K-ATPase activity after renal ischemia and gentamicin nephrotoxicity. An 18.6% reduction in renal cortical Na-K-ATPase activity was shown after 30 min of renal pedicle clamping and 60 min of reflow, and a 32.5% reduction after 90 min of single injection of 100 mg/kg gentamicin. Light and electronic microscopy revealed no significant morphologic changes in both groups in the experiment. 4g/kg FDP and 18g/kg Danshen were infused 60 min after reflow in the ischemic group, 90 min after injection of gentamicin in the gentamicin treated group and 90 min in the sham-operated group separately. The enzyme activity in the FDP-treated groups was found to be higher than that in the control kidneys while in the Danshen-treated groups no significant difference could be found. Our study showed that FDP and Danshen could prevent the decline of renal cortical Na-K-ATPase activity induced by ischemia and gentamicin. FDP could increase this enzyme activity while Danshen showed no such direct effect. PMID- 2560956 TI - Abdominal pain, distention and huge amount of ascites. PMID- 2560957 TI - Protective effects of sodium selenite on experimental myocardial infarction. AB - The present study compares the protective effects of sodium selenite (Se), hyaluronidase (Hy) and anisodamine (An) on infarct size, left ventricular myocardial contractility (LVMC) and relaxation (LVMR) and myocardial hypertrophy on the 3rd, 9th and 21st days after the ligation of left main coronary artery in the rats. The results showed that Se could reduce the infarct size, so could Hy and An. However, Se could relevantly improve LVMC and LVMR at the acute phase of infarction, while Hy and An could not. On the 21st day (healing phase) of infarction the indexes of the LVMC and LVMR in Se-, Hy- or An-treated rats were significantly better than those in the control rats. Se could enhance the extent of hypertrophy in non-infarcted myocardium, while Hy and An could not. On the 21st day of this experiment the total natural mortality in the Se-treated rats was significantly lower than that in the control or in the An-treated rats. These data suggest that Se is superior to Hy and An in the treatment of acute myocardial infarction. PMID- 2560958 TI - [Crevicular fluid myeloperoxidase and periodontitis disease]. AB - The volume and myeloperoxidase (MPO) activity of gingival crevicular fluid (GCF) collected with filter paper strips for 30 sec from the sulcus of healthy, gingivitis and periodontitis sites of Chinese subjects were measured. MPO/site and MPO/microliter GCF were both greater at gingivitis and periodontitis sites than healthy sites. Enzyme activity was similar at the 2 categories of diseased sites. Mean GCF volume and MPO activity of the samples were calculated for all sites with GI 0, 1, 2 and 0 + 1 irrespective of experimental group assignment. GCF volume MPO/site and MPO/microliter GCF all were greater at GI 2 than GI 0 or 0 + 1. These data indicate that increased GCF MPO previously observed at periodontitis sites is not specific to these sites. Rather, increased GCF MPO likely occurs when additional polymorphonuclear leukocytes enter the sulcus as a result of gingival inflammation. A second sample was obtained from 22 sites 4 weeks after the initial collection. These samples were collected for 5 rather than 30 sec. The GCF volume, MPO/site and MPO/microliter GCF each were greater in samples collected for 30 rather than 5 sec. Correlation coefficients showed that the amount of GCF and MPO activity of the fluid collected for 5 and 30 sec was dependent upon the site even though the samples were collected at different times. PMID- 2560959 TI - [Clinical application of artificial synthetic hydroxyapatite in the obturation of cavities of the jaw]. PMID- 2560960 TI - [Advances in recent clinical studies of liver cancer]. PMID- 2560961 TI - [An analysis of improved long-term survival of patients with liver cancer]. AB - Sixty-six patients surviving over 5 years after resection of primary liver cancer (PLC) were investigated. Of them, subclinical stage amounted to 56.1% (37/66), moderate stage 43.9% (29/66). There were 35 cases with small PLC(less than or equal to 5 cm). Radical resection was performed in 98.5% (65/66). Reoperation for subclinical recurrence and solitary pulmonary metastasis was done in 14 patients, and delayed resection of huge tumor was done in 3 patients. All patients were followed up from 60 to 319 months (mean, 115 months) up to the end of June 1988. Disease free survival was 80.3% (53/66) and death from recurrence was 19.7% (13/66). Authors discussed some aspects of improving Long-term survival of the patients. PMID- 2560962 TI - [Pathologic analysis of primary hepatocellular carcinoma in 19 cases surviving over 10 years]. AB - Pathological study was undertaken in 19 PHC patients who had survived over 10 years after operation. The pathological characteristics of the specimens were as follows: (1) Tumors less than 5 cm in diameter accounted for 78.9% (15/19), of which 13 PHC less than or equal to 3 cm in diameter. (2) Solitary tumors were found in 78.9% (15/19). (3) Sixteen out of nineteen (84.2%) cases had peritumor fibrous encapsulation, and 13 cases had fibrolamellar structures in cancerous foci. (4) 14 out of 19(73.7%) cases were found to have lympholeukocyte infiltration in cancerous foci or encapsulation. (5) Liver cirrhosis was found in only 42.1% (8/19) of all cases and 75% of them were slight cirrhosis. These were the pathological background accounting for long survival duration of PHC patients. The authors suggest that early detection, diagnosis, and treatment be done in this period, for better opportunity of survival in PHC patients. PMID- 2560963 TI - [Digital subtraction hepatic arteriography and venography]. AB - DSA and venography were performed in 13 hepatic carcinomas of central type for the location of the tumor. The procedure was found to clearly show the mass in its relation to hepatic veins. Preoperative demonstration of the landmarks of the hepatic lobes and segments was helpful to accomplish hepatic lobectomy or segmentectomy. The authors elaborated the technic and points for attention involved in this modality. The value of the hepatic arteriography and Venography in clinical application is discussed and indications are put forward. PMID- 2560964 TI - [Clinical study of the surgical treatment of hepatocarcinoma with an implantable microwave device]. AB - This article reports the surgical treatment to 51 patients with mid and advanced hepatocarcinoma by the self-made implantable microwave radiation apparatus from April, 1987 to April 1989. By way of histology, histochemistry, and electronic microscopy, we studied the killing effect on cancer cells, the range of the effect and the remarkable hemostatic mechanism of this implantable microwave device. Clinically, we developed six different methods of operate with the microwave. Without the need of blocking hepatic portal, this new approach simplifies the operation. The risk of iatrogenic cancerous diffusion and hemorrhage is also greatly reduced or prevented in hemihepatectomy. Initial results shows that the resection rate of the hepatocarcinoma went up from 20-42.5 percent to 74.5 percent. Microwave operation on 38 patients with advanced hepatocarcinoma reveals a survival rate of 75 percent over 6 months. The authors believe that this new operative method is effective in the treatment to advanced hepatic carcinoma, and prolongs the life of the patients. PMID- 2560965 TI - [Application of localized hyperthermia solidification combined with surgery in the treatment of liver cancer: a report of 34 cases]. AB - In 1980, we developed a Localized Hyperthermia Solidification Apparatus (LHSA), which can produce localized controllable hyperthermia and solidify the tumor tissues. Without the interference of the Corporal and visceral physiology. Animal experiments and pathological study proved it an effective and Safe means to deal with visceral cancers. From 1982 to 1988, 34 cases of liver cancer patients received the LHSA therapy combined with operations. The therapy was effective in 86.19% of cases, and the longest survival time was 5 years. PMID- 2560966 TI - [Microwave radiation treatment of liver cancer: animal experiment and preliminary clinical application]. AB - Microwave radiation treatment of liver cancer in animals with microwave treatment apparatus (2450 MHz) in either a Single-antenna or 8-antenna (combined) radiator and its clinical use were reported. In laparotomy, the antenna was inserted into liver tissue or the hepatoma. The changes of liver pathology and ultra structure as well as the process of absorption and healing of radiated liver were investigated. The results showed that a large amount of air bubbles were formed in liver tissue and its small veins when the temperature was higher than 80 degrees C in the radiated center. These bubbles were drastically reduced in number when the temperature was controlled around 65 degrees C. The radiated liver was pathologically divided into three parts: necrosis area (the center of radiation), normal area (far from the center) and reactive area (between the necrosis and normal areas). The largest diameter of the coagulated area was about 5 cm. The coagulated tissue was completely absorbed 60 days after the operation. Seventeen cases of primary hepatoma and 8 cases of secondary liver cancers were treated with microwave radiation therapy with satisfactory results. The volume of bleeding during the operation was greatly reduced. No complication was found. PMID- 2560967 TI - Development of the receptors to prostaglandin E2 in the rat kidney and neonatal renal functions. AB - Inhibition of prostaglandins (PGs) synthesis induces responses on renal function that are age-dependent. Indomethacin in the adult rat potentiates the response to vasopressin whereas in the newborn rat blocks the response to dehydration. These differences suggest that the sensitivity of the renal tissue to PGs changes as the animal matures. This study was designed to analyze the characteristics of the receptors to PGE2 in the developing rat and to compare with that of the adult rat. Binding was performed in cortical and medullary slices incubated with tritiated PGE2. It was found that the renal uptake of the radioactive ligand had three components, the first is the synthesis of endogenous prostaglandins, the second is tubular secretion (only in cortex) and the third is binding to receptor sites. We found it necessary to eliminate the first two components (with indomethacin and p-amino-hippurate) to achieve an adequate measurement of the apparent affinity of the binding sites to PGE2. Under these conditions the cortex bound more PGE2 than the medulla (at all ages) and the highest affinity was observed in the neonatal cortex. Synthesis of prostaglandin E2 was higher in newborn than in adult rats. In vivo, indomethacin and acetaminophen blocked the response of the newborn rat to dehydration whereas in the weaning and adult rat the response was not changed by these inhibitors of the synthesis of prostaglandins. Our results suggest a major role of the prostaglandins in the regulation of the water balance in the neonate. PMID- 2560968 TI - [The role of Na+/H+-exchange and diffusion of lactic acid in acid-dependent uncoating of influenza virus]. PMID- 2560969 TI - [Transposon Tn5-induced repair of genetic damage of replication of plasmid RP1 and hybrid derivative of RP1 and RP4 plasmids]. PMID- 2560970 TI - [Determination of a minimal segment of E. coli ribosomal protein L10 retaining its regulatory function]. PMID- 2560971 TI - [Expression of human tissue-type plasminogen activator cDNA in human cultured cells controlled by different eukaryotic promoters]. PMID- 2560972 TI - [Formation of covalently-bound circular structures of transposon TnV (Tn5-Rep pSC101) during transposition]. PMID- 2560973 TI - [Cloning of cDNA fragment of human brain kainate receptor]. PMID- 2560974 TI - [Organ specific deposition of 45Ca labelled calcium phosphate ceramic material]. AB - Research work using easily resorbable HA- und TCP-ceramics does not give any proof for pathologic deposition in different organs. Based on current information, there is no objection against the clinical use of HA-ceramics. PMID- 2560975 TI - Race, borderline hypertension, and hemodynamic responses to behavioral stress before and after beta-adrenergic blockade. AB - Blood pressure (BP) and hemodynamic responses (cardiac output [CO] and total peripheral resistance [TPR]) to a competitive reaction-time task, previously shown to increase beta-adrenergic activity, were compared in 20 Black and 20 White young men, once following infusion of saline placebo and later repeated after infusion of the nonselective beta-antagonist, propranolol. Both racial groups included subjects with marginal systolic BP (SBP) elevations (n = 6 Whites, n = 5 Blacks) and subjects with normal BP (n = 14 Whites, n = 15 Blacks). Blacks with marginal SBP elevations showed greater diastolic BP increases during the stressor than their White counterparts, both before and after beta-blockade. Both Blacks and Whites with marginal SBP elevations showed greater CO increases during the task than normotensives prior to blockade. Independent of BP status, Black subjects consistently showed higher TPR responses to the task than Whites (lesser decreases before blockade and greater increases after blockade). Blacks also showed greater diminution than Whites after blockade in ionotropic myocardial responses (stroke volume, pre-ejection period, and CO) to the stressor. The possible contributions of alpha-adrenergic and beta-adrenergic influences to these racial group differences are discussed. PMID- 2560976 TI - Differential cross-tolerance to opioid agonists in morphine-tolerant squirrel monkeys responding under a schedule of food presentation. AB - The effects of various mu and kappa opioid agonists were evaluated in three squirrel monkeys responding under a fixed-ratio 30 schedule of food presentation before, during and after a regimen of chronic morphine administration. Initially, dose-effect curves for the mu opioid agonists morphine and l-methadone, the kappa opioid agonists U50,488 and tifluadom, the mixed mu/kappa opioid agonist ethylketocyclazocine, and the non-opioid compound pentobarbital were determined in non-tolerant squirrel monkeys. Subsequently, monkeys were administered up to 3.0 mg/kg of morphine twice daily for 8-9 weeks, which resulted in a 1/2 to 3/4 log unit shift to the right of the morphine dose-effect curve relative to its prechronic position. During the chronic morphine regimen, the l-methadone dose effect curve shifted to the right approximately 3/4 log unit, while the U50,488 and pentobarbital dose-effect curves did not change. In contrast, the ethylketocyclazocine and tifluadom dose-effect curves shifted to the left approximately 1/4 and 3/4 log unit, respectively. The lack of cross-tolerance between mu and kappa agonists in morphine-tolerant squirrel monkeys observed in the present study provides further support for the differentiation of mu and kappa agonists. The occurrence of leftward shifts in the dose-effect curves of some opioid compounds with kappa agonist activity during the regimen of chronic morphine administration suggests that morphine tolerance modulates their PMID- 2560977 TI - Enhanced natriuretic potency of intravenous clonidine: extrarenal site of action? AB - We have previously demonstrated that low intrarenal infusion rates of clonidine selectively increased water excretion, whereas higher infusion rates were required to increase solute excretion. This is in contrast to previous experiments where intravenous administration of clonidine resulted in a concomitant increase in water and sodium excretion. We therefore determined the dose response curve for an intravenous infusion of clonidine on water and solute excretion and compared this to the effects of an intrarenal infusion. Uninephrectomized rats were anesthetized and the remaining kidney isolated for the collection of urine. Clonidine (0.1, 0.3, 1 or 3 micrograms/kg per min) or vehicle (saline) was administered either intravenously or intrarenally. Both intravenous and intrarenal administration of clonidine produced a dose selective dissociation of water and solute excretion, that is, at low infusion rates only urine volume was increased. Higher infusion rates were required to increase sodium excretion. In addition, intravenous administration of clonidine was more potent in producing a natriuresis, suggesting that the renal effects may be, in part, secondary to additional peripheral and/or central effects of this agonist following this route of administration. PMID- 2560978 TI - Evidence for the presence of 5-HT1-like receptors in rabbit isolated basilar arteries. AB - The 5-hydroxytryptamine (5-HT) receptor mediating contraction of rabbit isolated endothelium denuded basilar artery has been investigated. 5-HT and a variety of 5 HT receptor agonists contracted rabbit isolated basilar artery with a rank order of agonist potency: 5-carboxamidotryptamine (5-CT) greater than 5-HT greater than GR43175. None of these agonists relaxed rabbit isolated basilar artery when tone was elevated with prostaglandin F2alpha. The contractile response to both 5-HT and GR43175 was resistant to antagonism by GR38032, phentolamine, (+/-) cyanopindolol and yohimbine. Ketanserin (100 nM) and mesulergine (100 nM) produced small significant rightward shifts of C-E curves to 5-HT with respective concentration-ratio shifts of 5.7 (1.5-21.0 95% confidence interval and 2.89 (1.1 7.6 95% confidence interval). GR43175-induced contraction was resistant to antagonism by ketanserin however the maximum response to GR43175 was significantly reduced in the presence of mesulergine, with no change in EC50. Methiothepin was a potent antagonist of the contractile actions of both 5-HT and GR43175, with respective pA2 values against each agonist of 10.3 and 9.9. The slope of the Schild regression for methiothepin against 5-HT-induced contraction was significantly less than unity. Methiothepin (100 nM) had no effect on the contractile response to the thromboxane A2 mimetic U-46619. It is concluded that 5-HT and GR43175 contract rabbit isolated basilar artery by activating a 5-HT1 like receptor. In addition 5-HT may activate a population of 5-HT2 receptors producing a further contraction of rabbit isolated basilar artery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2560979 TI - A potent antagonist of the strychnine insensitive glycine receptor has anticonvulsant properties. AB - 5.7-Dinitro-quinoxaline-2.3-dione (MNQX) displaced [3H]glycine binding to cortical membranes but had no effect n [3H]3-((+/-)-2-carboxypiperazin-4-yl) propyl-1-phosphonic acid ([3H]CPP) binding. MNQX potently antagonized N-methyl-D aspartate (NMDA)-evoked release of [3H]GABA from cultured cortical neurones, NMDA evoked spreading depression and NMDA depolarizations in the rat neo-cortex. All of these responses were reversed by addition of glycine to the perfusion media. These results suggested that MNQX is an antagonist at the strychnine-insensitive glycine receptor associated with the NMDA receptor/ionophore complex. Furthermore the compound was found to antagonise audiogenic seizures in DBA-2 mice indicating the potential of glycine antagonists of this type in anticonvulsant therapy. PMID- 2560980 TI - Omega-conotoxin GVIA blocks synaptic transmission in the CA1 field of the hippocampus. AB - The effects of omega-conotoxin GVIA (omega-CgTx), a peptide isolated from the venom of a marine mollusc, were studied in rat hippocampal neurons. Intracellular recordings from the CA1 area were made for the purpose in in vitro slice preparations. Omega-CgTx (0.1-1 microM) rapidly and irreversibly blocked the EPSP and the IPSPs elicited by electrical stimulation of Schaffer collaterals/commissural fibers. Omega-CgTx also blocked the slow cholinergic EPSP induced by electrical stimulation of cholinergic afferents. The postsynaptic effects of baclofen or carbachol remained unchanged in the presence of omega-CgTx and other postsynaptic calcium-dependent events such as afterhyperpolarization were not affected by omega-CgTx. These results suggest a presynaptic action of omega-CgTx through the blockade of neurotransmitter release. Omega-CgTx might act in the hippocampus by blocking presynaptic N-type voltage-sensitive calcium channels. PMID- 2560981 TI - The use of mu opioid receptors in the mouse vas deferens for the study of opiate tolerance. AB - We have previously shown that mu opioid receptors in the guinea pig ileum preparations are poor models for those in the central nervous system to study the phenomenon of opiate tolerance. Since we have shown subsequently that the mu opioid receptors in the mouse vas deferens preparation differ from those in the guinea pig ileum, we investigated the possibility that the mu receptors in the mouse vas deferens might be better models for the study of opiate tolerance. The degree of tolerance produced by incubation of mouse vas deferens preparations with morphine in vitro was relatively low. However, when mice were either implanted with morphine pellets (75 mg free base) or injected s.c. with 100, 50 or 25 mg/kg of morphine sulfate, a large magnitude of tolerance in their vasa deferentia was consistently observed. When pA2 analyses were performed in the tolerant vasa deferentia (25 mg/kg) using naloxone, the pA2 value was 7.55 +/- 0.18 compared to the control value of 8.45 +/- 0.24. Since the tissue became significantly more refractory to the effects of the opiate antagonist with the development of tolerance, which is the opposite of what happens with central mu receptors, we conclude that the mu opioid receptors in the mouse vas deferens are not adequate models for those in the central nervous system to study the phenomenon of tolerance. PMID- 2560982 TI - Neuropeptide Y and peptide YY interact with rat brain sigma and PCP binding sites. PMID- 2560983 TI - Aromatase activity in cultured ovaries from fetal and neonatal golden hamsters. AB - The production of oestrogen from androstenedione by ovaries of hamsters on day 14 of pregnancy and on days 0 and 3 after birth was investigated by culturing these ovaries for 48 h in serum-free medium containing insulin and [3H]androstenedione. Both oestrone and oestradiol-17 beta were produced in culture of ovaries on day 14 of pregnancy. The production of oestrogen increased about 2-fold in culture of ovaries on days 0 and 3. Dibutyryladenosine 3':5'-cyclic monophosphate stimulated the oestrogen production by ovaries on day 14 of pregnancy and on days 0 and 3. The present results indicate that the ovaries of fetal and neonatal hamsters have an aromatase activity and that these aromatase activities are responsive to adenosine 3':5'-cyclic monophosphate. PMID- 2560984 TI - Age-related changes in adrenocorticoid and opioid receptor capacity of thymus derived lymphocytes in rats. AB - The experiments were performed in 4-day, 4-week, and 4-month old rats. The total binding capacity of high affinity receptors for 3H-naloxone and 3H-corticosterone in thymus-derived lymphocytes was measured in vitro. There was no change in the affinity constant of the receptors for the ligands during the life-time mentioned before. The maximal binding capacity for 3H-corticosterone in thymus-derived lymphocytes showed a marked increase in 1-month and 4-month old ages as compared to the values obtained by the end of the first postnatal week. In contrast, the maximal binding capacity of lymphocytes for 3H-naloxone showed a significant decline with age. Changes in the binding capacity for the two ligands refer to changes in composition of of cells within the thymus and to alterations in cell system in sensitivity to either corticosteroids or opioids. PMID- 2560985 TI - The dexamethasone suppression test and long-term contraceptive treatment: measurement of ACTH or salivary cortisol does not improve the reliability of the test. AB - Under the influence of high estrogen levels, the suppression of total serum cortisol in the dexamethasone test has often been found to be incomplete. Its measurement for the purpose of excluding Cushing's disease or adrenal tumors in women taking oral contraceptives is, therefore, considered unreliable. This study was designed to compare the reliability of measurements of total cortisol, unbound cortisol and ACTH suppression during chronic hyperestrogenaemia. An overnight suppression test with 2 mg of dexamethasone was performed in 19 women receiving long-term contraceptive treatment (group A) and in 12 controls (group C). Baseline and post-dexamethasone morning levels of ACTH, total serum cortisol and unbound salivary cortisol were determined by RIA. In addition, unbound serum cortisol was measured by equilibrium dialysis. Mean baseline levels of all four parameters were significantly higher in group A. This result points towards the possibility of a direct stimulatory effect of estrogens upon the corticotroph axis which is independent from CBG-mediated increase of total cortisol. Post dexamethasone values of total and unbound cortisol showed no statistically significant differences, while ACTH suppression in group A was even slightly better than in group C. From these data it is concluded that there is no need for post-dexa routine measurement of ACTH or unbound cortisol under contraceptive treatment since neither one of these parameters provides any additional information in comparison to total cortisol. PMID- 2560986 TI - Influence of electrical stimulation of the limbic structure on adrenocortical steroidogenesis in hypophysectomized rats. AB - The effects of electrical stimulation of the medial amygdala (AMYG) and dorsal hippocampus (DHPC) on the rates of 14C transfer from 14C-1-acetate into adrenocortical steroids in adrenal slices of hypophysectomized rats were investigated. The 14C transfer rates into corticosterone were increased by stimulation of the AMYG and DHPC. The 14C transfer rates into cortisol were increased by the AMYG stimulation but were not altered by the DHPC stimulation. From these results, it might be suggested that these limbic structures were involved in the regulation of adrenocortical steroidogenesis without participation of the pituitary. PMID- 2560988 TI - [Ultrastructural localization of leptospiral antigens by colloidal gold technique]. AB - The rabbit anti-mouse antibodies were successfully labelled with colloidal gold. The distribution of leptospiral antigens in ultrastructure was researched with McAbs and labelled antibodies. It was found that colloidal gold particles were mainly distributed at the outer membrane of leptospiral strain O17 cells, which indicated that the antigens recognized by McAbs 1A7E7, and 2F9D4 were localized at the outer membrane of leptospiral cells. It was thought that colloidal technique would provide a method for research on the antigen distribution of leptospiral cells in ultrastructure. PMID- 2560987 TI - Effects of training on regional substrate oxidation in the hearts of ageing rats. AB - 23-month-old male rats were trained by running for 20 weeks. The oxidation rates of succinate, glutamate+malate, palmitoylcarnitine, and pyruvate and the activities of lactate dehydrogenase, citrate synthase, isocitrate dehydrogenase and cytochrome oxidase were measured in the subendocardium and subepicardium and in the right ventricle. Regional differences of substrate oxidation rates in the myocardium of old sedentary or trained rats were less than in young rats, suggesting that regional differences in the cardiac work load disappear during ageing. Training did not improve oxidation rates, in contradiction to some previous results. PMID- 2560989 TI - [Diagnosis of congenital CMV infection by specific CMV IgM antibody in cord serum with ELISA]. AB - Cytomegalovirus IgM (CMV IgM) antibodies of 582 cord sera from 6 hospitals in Chengdu were detected by direct ELISA and indirect ELISA. Twenty-seven cases with CMV IgM antibody positive were detected from 582 cord sera. Twenty-five and 15 cases were found to have CMV IgM antibody by direct ELISA and indirect ELISA, respectively (P greater than 0.05). Indirect ELISA for detection CMV IgM antibody was only interfered by high concentration of CMV IgG and rheumatoid factor. Children with CMV IgM antibody positive were followed-up to 5-6 months. Hearing loss was detected by impedance audiology in only 2 cases. But the other physical and mental examinations were normal. The result showed that the prevalence of congenital CMV infection is 4.6% in our study and direct ELISA method is more specific and sensitive than indirect ELISA method to detect CMV IgM antibody in cord serum. PMID- 2560990 TI - Clear cell sarcomas of the tendon sheath. An experience of 22 cases seen over 16 years. AB - Twenty-two histologically confirmed cases of Clear Cell Sarcomas (C.C.S.) seen over a sixteen year period, are retrospectively analysed from a clinico pathological view-point. The tumours occurred in young adults, average 36 years. Males predominated, and the lower extremity particularly the thigh and foot were the most commonly involved. Grossly, a connection with aponeurosis and tendon sheath has not been recorded consistently. Microscopically the tumour was distinctive, with their short fascicles and nests of polygonal to fusiform cells. The "clear" cytoplasm stressed in it's very name was not the rule. Melanin was demonstrated in 7 of the 22 cases, i.e. 32%, and these patients did not behave any differently from those where pigment was not seen. Immunocytochemistry for S 100 protein was done in 14 cases and was positive in 13 cases. Follow-up information revealed that 11 patients were dead of advanced disease in an average period of 24 months. Two patients were alive with disease for an average period of 3 years 7 months. Only 4 patients had no evidence of disease for periods ranging from 6 months to 26 months. Metastasis to nodes was a very frequent event, but lung and bony metastasis has also been documented. PMID- 2560991 TI - Extrauterine mixed mesodermal tumour of ovary with heterologous element. AB - The ovary is very rare site of mixed mesodermal tumour. The present case occurred in postmenopausal woman aged 52 years, in right ovary with endometrioid carcinomatous component admixed with chondrosarcomatous component and squamous element. The patient was disease free at one year follow up, with no evidence of local recurrence and distant metastases and was treated post-operatively with combination therapy of radiotherapy and chemotherapy. The review of literature is discussed briefly, with its histogenesis and its differential diagnosis from immature teratoma, malignant teratoma, sertoli leydig cell tumour containing islands of cartilages and embryonal carcinoma of ovary. PMID- 2560992 TI - CpG frequency-dependence in the activity of eukaryotic DNA methyltransferase: in vitro methylation of CpG-rich islands. PMID- 2560993 TI - Inosine kinase activity from cultured Chinese hamster lung fibroblasts (V79). PMID- 2560994 TI - Purine salvage enzyme pattern in human intestinal carcinoma. PMID- 2560995 TI - [Retinoid-receptor proteins]. PMID- 2560996 TI - [Wilms' tumour: chromosome findings and carcinogenesis]. PMID- 2560997 TI - [Chromosome and molecular genetics--molecular analysis of aniridia-Wilms' tumor syndrome]. PMID- 2560998 TI - [Acute myeloblastic leukemia development in a patient with small cell lung cancer in complete remission]. AB - A 67 year-old man was admitted to our hospital because of cough and sputum. He smoke one pack of cigarettes a day for more than twenty years and the chest X-ray film revealed a mass in the left hilum and left sided pleural effusion. The diagnosis of small carcinoma of the lung (limited disease, T4N1MO, stage 3B) was made by trans-bronchial lung biopsy and radiographic studies. Both chemotherapy (nimustine (ACNU), cyclophosphamide, vincristine, and methotrexate) and radiation therapy was started, however, the chemotherapy was discontinued in July 1987 because of severe anemia. The diagnosis of refractory anemia with excess of blasts in transformation (RAEB in T) was made by bone marrow aspiration and the patient was treated by transfusion (400-800 ml/week). In December 1987 transition to acute myeloblastic leukemia was confirmed by another bone marrow aspiration biopsy and the patient was given low dose cytosine arabinoside (Ara-C). The response was favorable in the beginning but in about two months pancytopenia became refractory and the patient died in June 1988. Clinically there was no sign of local or distal recurrences of lung cancer, and the complete remission of small cell lung cancer (SCLC) was confirmed by autopsy. Survival in SCLC remains poor, so that the choice of treatment is still the primary concern, however, development of other malignancies which include acute leukemia is another problem which should be taken into account when the treatment is extensive. PMID- 2560999 TI - Hepatitis B virus DNA integration in hepatocellular carcinomas and their adjacent non-neoplastic liver tissues. AB - Hepatitis B virus (HBV) DNA integration was studied in 24 hepatocellular carcinoma (HCC) tissues obtained at operations or autopsies. In 11 cases whose sera were positive for hepatitis B virus surface antigen (HBsAg), HBV DNA integration was demonstrated by Southern blot analysis. Only one of 6 cases whose sera were positive for hepatitis B surface antibody (HBsAb) but negative for HBsAg revealed the integration and the other 5 cases revealed no HBV DNA integration. HBV DNA amplification was noted in 4 of these 6 cases in which HBV DNA integration was found when compared with the adjacent liver tissues. The integration pattern of HBV DNA was different in one case between primary HCC tissue and a metastasized lymph node. It is suggested that HBV DNA amplification is not directly related to the development of HCC and that there are polyclonal tumor cells which have different patterns of virus genome integrations. PMID- 2561000 TI - Treatment of pain according to syndrome differentiation in 169 cases of liver cancer. AB - In 169 cases with liver cancer pain, the therapeutic effects of TCM and the authors' "analgesic therapy of four steps" were analysed. The results were that most patients with advanced liver cancer had mild pain, and only a small number of patients (14.2%) had moderate or severe pain. By the first step analgesic therapy, 46.1% of the patients were relieved and with the second or third step of analgesic therapy, 93.5% of the patients were relieved, only 6.5% of the patients with grade III pain needed the fourth step therapy. With TCM treatment alone, the remission rate of grade I and grade II pain was 100% and 76.9% respectively. "Analgesic therapy of four steps" produced less side effects or addiction to narcotics. PMID- 2561001 TI - Endogenous virus genomes become hypomethylated tissue--specifically during aging process of C57BL mice. AB - In an attempt to find out a cause for age-dependent derepression of endogenous viruses, extents of DNA methylation at the endogenous B- and C-type ecotropic viruses in brain, liver and spleen of C57BL/6NJc1 were examined at three ages, newborn, young adult and old. Both endogenous viruses showed a slight but significant tissue-specific either hypo- or hypermethylation during post-natal developmental phase in the three tissues. After maturation, however, no such change was detectable at most of the sites examined. The exceptions were C-type ecotropic virus in brain and B-type virus in spleen, where the age-dependent decreases of methylation were observed. The changes seemed to be continuations of preceding developmental hypomethylation. They indicated that the hypomethylation could be one of the causes for the age-dependent derepression of endogenous virus. It was further suggested that a mechanism to stop the developmental changes of DNA methylation at the maturation of individuals would be important in considering the reasons for the changes in senescent phase. PMID- 2561002 TI - Age-related changes in the galactose recognition system in rat liver cells. AB - In the present report, the galactose recognition system in 3- and 24-month-old rat livers has been studied with in vitro and with in situ binding experiments and in vivo ligand uptake. The galactose-specific receptors were visualized by using colloidal gold particles of different sizes (5 nm, 17 nm and 50 nm mean diameter), coated with lactosylated bovine serum albumin (LacBSA) as electron dense ligands. The data show that aging affects the expression of galactose specific receptors and the rate of endocytosis. In the in vitro and in situ experiments hepatocytes and liver macrophages from old rats on the plasma membrane express a decreased number of binding sites with respect to those present on the adult rat liver cells. Approximately 80% of the total number of liver macrophages from aged rats show a binding distribution which is very different from the typical clustered receptor arrangement: the binding sites appear as single or small clusters of gold granules. As a direct consequence of the altered pattern of the receptor distribution, the capacity of liver macrophages from 24-month-old rats to internalize the larger ligands (17 nm and 50 nm) is decreased, as compared with adult rats. Aging, therefore, influences the galactose recognition system in two ways: (i) by decreasing the number of binding sites expressed on the liver cell surfaces; and (ii) by modifying the receptor distribution on liver macrophages and consequently affecting the internalization of galactose exposing particles. PMID- 2561003 TI - Age-related changes in protein phosphorylation by rat hepatocytes. AB - The qualitative and quantitative changes in the phosphorylation of specific proteins in hepatocyte suspension from 5-, 12- and 22-month-old male Fischer F344 rats were analyzed by two-dimensional polyacrylamide gel electrophoresis. No qualitative changes in phosphorylation of individual proteins were observed with age. In addition, very few quantitative changes (less than 2% of proteins studied) in protein phosphorylation were detected. The phosphorylation of two acidic proteins decreased (50%) with age while the phosphorylation of one basic protein increased (300%) with age. The two acidic proteins and one basic protein that showed quantitative changes with age were found predominately in the microsome and nuclear fractions of hepatocytes, respectively. The effect of dietary restriction on the phosphorylation of proteins in male Fischer F344 rats was also studied. Although, dietary restriction alters the age-related incident of disease and prolongs longevity, it did not have any significant effect on phosphorylation of individual proteins in the liver. PMID- 2561004 TI - Effect of passive transfer of myasthenic serum on mechanical, electrical and neuromuscular transmission properties of mouse skeletal muscle. AB - Neuromuscular transmission, muscle electrical activity and muscle mechanical properties have been studied in mice after the intraperitoneal transference of sera from myasthenic patients. Measurements of mepp's amplitude appear to be a suitable method for myasthenia gravis diagnosis since a high percentage of the sera from these patients caused a decrease in mepp's amplitude. The increase of the premenstrum weakness in myasthenic patients did not appear to be associated with greater fall of mepp's amplitude, since serum obtained in the premenstruum and far from it produced similar results. Myasthenic serum did not modify the electrical properties of innervated muscle sarcolemma. The resting membrane potential and the action potential parameters remained unchanged. However, tetrodotoxin resistant-action potentials of denervated muscles was reduced by myasthenic serum, possibly in association with receptor endocytosis process induced by the immunoglobulin. Muscle mechanical properties did not show alterations, even in the presence of positive titer of anti-striated muscle antibody. PMID- 2561005 TI - Lipid peroxide and antioxidant enzymes in muscle and nonmuscle of dystrophic mouse. AB - To determine whether abnormality in redox metabolism occurs specifically in certain individual dystrophic muscles, thiobarbituric acid reactivity, free radical scavengers, and oxidative marker enzymes were measured in the liver, kidney, erythrocytes, heart, and four different individual skeletal muscles from C57BL/6J dy/dy mice. Superoxide dismutases were assayed by specific radioimmunoassays, which enabled the study of a small individual murine muscle. Glutathione peroxidase and catalase were increased markedly in each individual dystrophic skeletal muscle studied and less markedly in the heart. Manganosuperoxide dismutase and thiobarbituric acid reactivity were decreased to a similar extent in each dystrophic skeletal muscle. Cuprozinc superoxide dismutase was decreased in the soleus muscle. Only a minimal biochemical change occurred in nonmuscles. Fumarase activity correlated closely with the level of manganosuperoxide dismutase. These results suggest that muscle protein breakdown occurs independently of lipid peroxidation despite the presence of tissue specific abnormality of redox metabolism in dystrophic muscle. PMID- 2561006 TI - [Regulation of the postsynaptic alpha-1-adrenoreceptors of the smooth muscle of the nictitating membrane in the cat]. AB - Radioassay binding of the smooth muscle of the nictitating membrane of cat has revealed the specific binding sites of [3H]prazosin corresponding to alpha-1 adrenoceptors. There was a significant increase in the number of alpha-1 adrenoceptors without any changes in their affinity. Incubation of the culture of the preliminary sympathectomized smooth muscle with noradrenaline decreased the number of alpha-1 adrenoceptors also without altering the affinity of binding. So, it is concluded that sympathetic nervous system regulated the number of postsynaptic alpha-1 adrenoceptors by means of neurotransmitter (noradrenaline). PMID- 2561007 TI - [The effect of cadmium ions on synaptic transmission in the tectum of the frog]. AB - The effect of cadmium ions on the synaptic transmission in the frog tectum was analyzed in acute experiments by the method of recording of the quantum of EEG (the extracellular monosynaptic potential of synapses of a single axon) [1]. It was shown that perfusion of the tectum with CdCl2 in the concentration of 10-200 mumol/l reversibly inhibited the quanta of EEG, decreased their duration and increased the synaptic delay. Consequently, cadmium in industrial waste may be dangerous regarding the harm to the CNS, too. The results of the presented work correspond to the supposition based on experiments in vitro that presynaptic potential-dependent calcium channels prove to be one of the areas of synaptic action of Cd++ ions. PMID- 2561008 TI - [The action of strontium and barium ions on the calcium binding and transport systems of nerve cells]. AB - Using Ca-sensitive fluorescent probe (fura-2) Sr and Ba absorption by intracellular organelles after cell loading by these cations and their effect on Ca release from intracellular stores were studied on isolated snail neurons. It was shown that Sr could effectively replace intracellular Ca, whereas Ba ions are not absorbed inside the cell and block a Ca-induced Ca release from intracellular stores. PMID- 2561009 TI - Cardiac beta-adrenergic sensitivity in depression: relation with endogenous subtype and desipramine response. AB - The authors studied the responsiveness of cardiac beta-receptors to isoproterenol, a noradrenergic agonist, in 29 depressed patients and 13 control subjects. They showed a significantly lower sensitivity in depressed patients as compared with the control subjects. Focussing on the group of depressed patients without antidepressant treatment in the month preceding the study (n = 15) in order to avoid a bias, the following significant results were obtained: cardiac beta-adrenergic receptor sensitivity was lower in patients suffering from endogenous depression than in those suffering from reactive depression (as classified by Newcastle Scale). There was a negative linear relation between cardiac beta-adrenergic sensitivity and the posttreatment clinical state (as expressed by the MADRS score) for the 9 patients who ended a 3-week desipramine treatment period. PMID- 2561010 TI - Cortisol, ACTH, prolactin and beta-endorphin responses to fenfluramine administration in major-depressed patients. AB - In the past, some researchers found increased cortisol and prolactin responses to the administration of fenfluramine in major-depressed patients. It was believed that the fenfluramine test could prove to constitute another challenge probe to reflect the central serotonergic function. The present study was conducted in order to investigate the pituitary/adrenal responses to fenfluramine in major- versus minor-depressed patients. To this end we administered 60 mg D,L fenfluramine p.o. to 40 depressed patients categorized according to the DSM-III. The basal levels of cortisol, adrenocorticotrophic hormone (ACTH), beta endorphins and prolactin and their levels 2 and 4 h after fenfluramine administration were measured. We found no significant effect for fenfluramine treatment on cortisol, ACTH or beta-endorphins. There was a significant (p = 0.02) effect for fenfluramine treatment on prolactin. The enhanced secretion of prolactin was only significant (p = 0.006) in major (296.X2, 296.X3, 296.X4) and not in minor (300.40, 309.00) depressives. It was concluded that our findings corroborate the thesis of a hypersensitive serotonergic neurotransmission during a major depressive episode. PMID- 2561011 TI - Effect of drugs acting on monoaminergic and cholinergic systems on the quantified EEG of rats. AB - Five different classes of drugs (clonidine: 0, 0.10, 0.30, 0.50 mg/kg; yohimbine: 0, 2, 4, 8 mg/kg; haloperidol: 0, 0.02, 0.04, 0.08 mg/kg; piracetam: 0, 150, 300, 600 mg/kg, and eserine: 0, 0.10, 0.30, 0.50 mg/kg) were studied on two cortical EEG derivations as well as a deep structure, the locus ceruleus in the rats. Each drug affected the EEG in its own particular manner. Clonidine significantly decreased frequency in the theta band (3.7-7.5 Hz) and increased it in the alpha band (7.6-13.5 Hz). A general significant increase in power was observed. Yohimbine's effects on the EEG varied according to the regions studied. There were significant modifications of power and frequency in the theta, alpha and beta bands in the three derivations studied. Notable effects of haloperidol were observed as an increase in power in all frequency bands at all doses administered particularly in the anteroparietal and posteroparietal derivations. In the locus ceruleus derivation, power was significantly increased at all doses only in the alpha and beta frequency bands. Concerning piracetam, while no significant effects were noted on the EEG frequency, this drug significantly increased EEG power in the posteroparietal and anteroparietal derivations. The most important effects are obtained at the lowest dose (150 mg/kg) administered. Finally, it was shown that eserine significantly decreased power in the delta bandwidth shortly after its administration. Afterwards, the power gradually regained its original level. PMID- 2561012 TI - [Osteosynthesis using polyglycolide fixation devices]. AB - The authors analyse their experience in using screws and rods made of a biocompatible resolving high molecular polymer polyglycolid (produced in the USSR) in the surgical treatment of some kinds of fractures in 30 patients. The healing of the fractures took place within the normal terms in 27 patients, there was slow consolidation in 2 patients and a false joint in 1 patient. The resolution of conical rods takes place after 6 months, and that of the screws after 3 years, with complete restoration of the bone structure at the implantation site. PMID- 2561013 TI - [Mapping and selective expression of nuclear antigens EBNA 3, 4 and 6 of Epstein Barr virus]. PMID- 2561014 TI - [Role of a purified Sertoli cell protein (CMB-21) in the biosynthesis of Leydig cell testosterone in the immature rat]. AB - The existence of Sertoli cell factors which modulate the rat Leydig cell function prompted us to study the biological activity of selected proteins called CMB proteins and produced by immature rat Sertoli cells. Percoll purified Leydig cells (10(5)) from 20 days-old rats have been incubated 5 h at 32 degrees C in 1 ml Ham F12/DME medium with increasing concentrations of partially purified CMB proteins (0-1,000 ng/ml) either in presence or absence of oLH (25 ng/ml). Among the CMB proteins tested, only CMB-21 produces a dose related increase of testosterone production: from 2 to 500 pg/ml of CMB-21, testosterone output is unchanged (51 pg/10(5) cells) but 1 to 1,000 ng/ml of this protein produces a linear increase of testosterone productions (86 to 870 pg). In the presence of oLH which induces a 10-fold increase of testosterone production (499 pg), increasing doses of CMB-21 further stimulate testosterone output (775 to 2.272 pg/10(5) cells). Whatever the concentration of oLH used (0 to 50 ng/ml), CMB-21 (500 ng/ml) leads to a further 2 fold augmentation of testosterone synthesis; similarly, in the presence of dbcAMP (1 mM), CMB 21 increases the testosterone production but no effect is observed when Leydig cells are incubated in the presence of 22R-hydroxycholesterol (30 microM). The cAMP levels which are increased more than 4 fold by oLH, remain unchanged in the presence of CMB-21 either alone or with oLH, as observed when Sertoli cell culture medium is used.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561015 TI - [Atrial natriuretic peptide response to physical exercise is inhibited by an antagonist of the opioid receptors]. AB - It was been shown that physical exercise increases plasma atrial natriuretic peptide (ANP) level. This effect was attributed to the hemodynamic changes of exercise which could increase atrial volume and result in ANP secretion. On the other hand, it was evidenced that morphine and opiate peptides greatly stimulate ANP release. To evaluate to what extent the endogenous opioid secretion during exercise induces the ANP release, six healthy volunteers male trained subjects were submitted to two maximal exercise tests with and without (placebo) opiate receptors blockade by naltrexone (50 mg per os). Blood samples were drawn before (rest) and after maximal exercise in order to measure by radioimmunological methods human atrial natriuretic peptide (alpha-h-ANP), beta-endorphin, plasma aldosterone (ALD), plasma renin activity (PRA) and corticotrophin (ACTH). Expired gas was collected during exercise to measure oxygen consumption. Subjects reached the same value of maximal oxygen consumption (VO2 max) at the end of exercise whatever treatment. Plasma ANP level at rest decreases slightly after administration of naltrexone (32.8 +/- 6.3 pg/ml with placebo versus 21.3 +/- 4.6 pg/ml with naltrexone) but the response to physical exercise was significantly reduced by naltrexone (73.3 +/- 14.9 pg/ml with placebo versus 46.9 +/- 8.6 pg/ml with naltrexone) (p less than 0.05). There was no statistical difference according to the treatment between the plasma levels of beta-endorphin, PRA and ACTH at rest as well as at the end of a maximal exercise.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561016 TI - [Effect of drugs with a known mechanism of action on catecholamine receptors on aggressive behavior in rats following administration of isoniazid]. AB - The participation of catecholaminergic neurones in effect of INH on affective aggression was studied using agents known to act on noradrenergic (clonidine, prazosin) and dopaminergic receptors (sulpiride, spiroperidol). The above mentioned agents blocked affective aggression in rats after administration of INH. The results of this study imply that both catecholamines participate in INH induced affective aggression. PMID- 2561017 TI - [Liver transplant in children. II]. AB - The family physician has a central role in preparing patients for transplantation and in the follow up. In the preoperative period adequate nutritional, vitamin intake and vaccinations are the mainstay of medical therapy. Pretransplant evaluation in a Liver Transplantation Centre is an essential part of the procedure, permitting an assessment of candidate's suitability and their position on the emergency list. During or immediately after the operation immunosuppression starts, according to personalized schedules and protocols. Among various complications, rejection must be promptly identified and treated. Resumption of a normal life style is the best indicator of a successful transplantation. PMID- 2561018 TI - Liposome-mediated increase of the superoxide dismutase content in human erythrocytes: characterization by electron spin resonance. AB - Room temperature electron spin resonance (ESR) spectroscopy, together with enzyme activity measurements, have been used to study the liposome-mediated enrichment of human red blood cells with superoxide dismutase. The ESR technique was found to be useful not only for qualitative and quantitative determinations, but also for demonstration of enzyme activity inside intact cells. The molecular properties of superoxide dismutase in relation to pharmacological and biotechnological applications are reviewed. PMID- 2561019 TI - Isolation and properties of lipolysis inhibitory proteins from wheat germ and wheat bran. AB - Proteins inhibiting pancreatic lipase in vitro have been isolated from wheat germ and wheat bran, with relative molecular mass ranging from 24,400 to 27,500. Inhibition of pancreatic lipase by the wheat germ proteins is related to their ability to interact with the emulsified substrate and to hinder the adsorption of the enzyme on the interface. The extent of inhibition depends on the amount of substrate and is independent of the enzyme concentration. Bile salts forming micelles in the concentration range used are able to partially reverse the inhibition of pancreatic lipase by the wheat germ proteins. The nutritional significance of the data obtained is discussed. PMID- 2561020 TI - [Effect of ACTH, dexamethasone, and adrenalectomy on the secretion of testosterone in the rat]. AB - The effect of ACTH (100 micrograms/animal/day, i.p.), dexamethasone (75 micrograms/animal/day, s.c.), both for three consecutive days, and adrenalectomy, with or without dexamethasone, maintained according to the group, one, two or three days, on the plasmatic testosterone and corticosterone levels, has been studied in adult male Wistar rats. ACTH and adrenalectomy produced a high decrease in testosterone levels (p less than 0.001 for the three days studied). Dexamethasone produced lower testosterone levels in the first day followed by partial recuperation between the second and the third days of its administration. Dexamethasone produced the effects mentioned for intact animals. The changes in corticosterone levels were according to an adequate response of the hypothalamus pituitary-adrenal system under these experimental circumstances. ACTH exerts an inhibitory effect on testosterone secretion in the rat, so that such an effect from the data obtained after adrenalectomy and simultaneous dexamethasone injections, does not seems to be mediated either by the presence of adrenals or high corticosterone levels. PMID- 2561021 TI - Effect of reducing agents and uncouplers on the electrical potential generated by mitochondrial ATPase activity. AB - Beef heart submitochondrial particles bound to phospholipids impregnated filters generated an electrical potential upon the addition of ATP. The magnitude of the electrical potential reached depended on the phospholipid mixture composition used for filter impregnation, phosphatidylethanolamine being the active component for the electrical potential generation. Uncoupler FCCP (p-trifluoromethoxy carbonyl cyanide phenylhydrazone) inhibited the transmembrane electrical potential generation by diminishing the electrical resistance of the system as a result of its protonophoric action. However, uncouplers 2, 4-dinitrophenol and dicoumarol did not provoke large modifications of the electrical resistance under the conditions of pH and concentration used, and their action varied with the time elapsed after the submitochondrial particles purification, favouring the idea of the uncoupler interaction with a specific site on the membrane. Addition of sodium dithionite resulted in a higher plateau value for the electrical potential consistent with the promoted increase in ATPase activity. The effect of this agent was reversed by the 2,6-dichlorophenol-indophenol added at equivalent concentrations. PMID- 2561022 TI - [Role of prostaglandins in the antihypertensive effect of the converting enzyme inhibitor, enalapril]. AB - Angiotensin converting enzyme (ACE) generates angiotensin II and is also capable of degrading bradykinin into inactive peptides. It has been suggested that the effects of ACE inhibitors are partially mediated by increased prostaglandin synthesis induced by a simultaneous rise in bradykinin. Captopril increases PG excretion and indomethacin (INDO) attenuates its effects. Enalapril is a long acting ACE inhibitor, and its molecule does not have the sulphydryl group present in captopril. In order to evaluate the participations of PG in a the ENA effects of enalapril (ENA) on arterial pressure (AP), plasma renin activity (PRA), plasma aldosterone (ALDO) and renal hemodynamics (RH) in essential hypertension (EHT), we compared the effects of ENA alone and associated with INDO. Nine EHT patients received on different occasions: ENA 10 mg, INDO 25 mg and ENA-INDO. Arterial pressure, PGE2, ALDO, PRA, RH and plasma and urinary ENA as enalaprylate were measured after each treatment. Maximal ENA absorption occurred after 4 hours, however it was still detectable after 72 hr. ENA decreased AP after 6 hr in spite of unchanged PGE2 excretion; PRA did not change and ALDO decreased transiently. INDO delayed ENA absorption, slightly attenuated the fall in AP and suppressed PGE2 excretion when given with ENA. INDO alone suppressed PGE2 and did not alter AP. No significant changes occurred in RH with the treatments. Our results suggest that the antihypertensive effect of ENA is independent of PG, and that the slight attenuation induced by INDO may be attributed to a delay in intestinal absorption. In EHT patients under normovolemic conditions, renal function is not altered by ACE inhibition. PMID- 2561023 TI - Mildly virilizing adrenal adenoma excreting large amounts of androgen metabolites. PMID- 2561024 TI - Synchronous colonic adenocarcinoma and extragenital malignant mixed mesodermal tumour. AB - A synchronous occurrence of large bowel adenocarcinoma and extragenital malignant mixed mesodermal tumour (MMMT) is reported. This case represents the sixth extragenital MMMT reported in the literature. PMID- 2561025 TI - [Extratesticular germinal cell tumors]. AB - In a group of 212 patients with germinal tumours of the testicle treated at the urological clinic in Hradec Kralove in 1975-1987 there were 7 with an extragonadal tumour in the retroperitoneum. In all the dominating symptom was a palpable resistance in the epigastrium, backache and secondary signs of malignant disease. In the first four a radical extirpation of the tumour was performed, in one with replacement of the aorta always with left nephrectomy. After introduction of cisplatinum into combined chemotherapy this became the initial treatment. Of the seven patients two died from generalization of the basic disease, the third one from exsanguination through an erosion of an aortal graft. PMID- 2561026 TI - Post hospital depression and the elderly cardiac patient. AB - This paper reports the findings of a modest study seeking to document the mood state of older patients post discharge from an acute medical facility following cardiac care. Our purpose was to identify factors which impede successful entry and participation of older patients in cardiac rehabilitation programs so modifications in program design can be made which would maximize successful outcomes for the elderly patient. PMID- 2561027 TI - Intraocular surgery and the repair of retinal detachments from AIDS-related diseases. PMID- 2561028 TI - Generic vs. proprietary otic drops. PMID- 2561029 TI - [Binding of latrotoxin to synaptosomes and synaptic membranes of the rat brain]. AB - The binding of [125I] alpha-latrotoxin to synaptosomes from the rat brain is studied. It is shown that the constant rate of toxin association with the synaptosome receptor at 37 degrees C is equal to 8.2 +/- 1.3 x 10(7) M-1.s-1, while that of synaptosomal membrane -7.6 +/- 2.7 x 10(6) M-1 s-1. Depolarization of the synaptosome membrane induced by 55 mM KCl decreases the binding rate of toxin to the receptor, the rate constant being equal to 3.9 +/- 1.5 x 10(7) m-1 s 1. The pattern of the dissociation process of the toxin-receptor complex of synaptosomes and of synaptosomal membrane is different. In the first case dissociation follows two stages with the rate constants 3.6 x 10(-3) s-1 and 1.2/10(-4) s-1, in the second case it follows one stage with the constant equalled 2.0 x 10(-5) s-1. The quantity of the toxin binding sites on synaptosomes may vary under the action of agents modifying the activity of calcium fluxes which are induced by alpha-latrotoxin. It is supposed that a decrease in the ATP level in synaptosomes as well as deenergy of the surface membrane leads to a change in the state of the alpha-latrotoxin receptor. PMID- 2561030 TI - [Formation of p-tyramine from dopamine bound to synaptic receptors of the rat brain in vitro]. AB - Tyramine (TA) revealed earlier during the functioning of dopamine (DA)-receptors of the rat brain (after learning) in vivo was produced from dopamine bound by DA receptors of the synaptic membranes in the system which was exposed to the influence of the microdischarge electroradialysis in vitro. It is shown that the formation of p-TA under these conditions depends on the period of the micro discharge effect on the system, it is maximal at exposition of 30 s for I = 4.2 mA. In control solutions of standard DA and DA preincubated with the membranes of the cerebellum homogenate, without DA-receptors, p-TA was not revealed under these conditions. The results obtained confirm the supposition that p-TA is the product of the DA-receptors functioning in vivo. PMID- 2561031 TI - [Beta-adrenoreceptors in the muscular tissue of Anodonta cygnea]. AB - The beta-adrenoceptors in the muscular tissue of Anodonta cygnea have been studied for the first time with the use of antagonist [125I] iodocyanopindolol. The tissue membrane had only one class of binding sites with Kd 2.9 +/- 0.02 pM and the maximal number of binding sites (Bmax) 110 +/- 2.4 fmoles/mg of protein. The potency of beta-agonists and antagonists for displacing [125I] iodocyanopindolol for its beta-AR complexes was the following: isoproterenol greater than adrenaline greater than noradrenaline-propranolol greater than serotonin much greater than dopamine greater than phentolamine. The GTP negative regulation of beta-AR affinity has been found. The data obtained show that the beta-AR are functionally coupled with GTP-binding protein which were similar to GTP-proteins of vertebrates. PMID- 2561032 TI - [Molecular mechanism of calcium ion transport in mitochondria. I. Glycoprotein peptide complex as a component of the electron transport system]. AB - The molecular structure of the mitochondrial glycoprotein capable of forming Ca2+ selective and ruthenium red-sensitive conductance channels when incorporating into a model membrane are studied. The glycoprotein is shown to be a complex consisting of the glycoprotein itself and a low-molecular component which may be attributed to the substance of a peptide nature. A technique is elaborated to divide the complex into constituents. It is found that the channel-forming part of the complex is its peptide component. The glycoprotein component is unable to transport Ca2+ and, probably, fulfills a regulatory function. PMID- 2561033 TI - [The effect of muramyl dipeptide analog, GMPD, incorporated into liposomes of various composition on the functional activity of macrophages]. AB - Liposomes of different composition and N-acetylglucosaminyl-N-acetylmuramyl-L alanyl-D-isoglutamine (GMPD) encapsulated in them are studied for their effect on the functional activity of macrophages by means of determining the 5' nucleotidase and adenosine deaminase activity in the in vivo experiments. It is shown that both liposomes and GMDP encapsulated in them increase the activity of adenoside deaminase and decrease that of 5'-nucleotidase. This evidences for a change in the adenosine metabolism in the alveolar and peritoneal macrophages and an increase in the functional activity of cells which resulted from that rise. The manifestation of the process depends both on the lipid composition of liposomes and their charge. The observed increase in the functional activity of the alveolar macrophages under the effect of liposomes and GMDP encapsulated in them correlates with inhibition of the lung metastases development in mice. PMID- 2561034 TI - Hepatic enzyme activities and plasma insulin concentrations in diabetic herbivorous voles. AB - The activities of the hepatic glycolytic enzymes glucokinase (GKase) and hexokinase (HKase) in herbivorous Microtus arvalis were very low and the hepatic fructose-1,6-diphosphatase (FDPase) activities were almost the same as those in C57BL/6J mice. Glycosuria was observed in over 50% of voles fed on a low fibre, high concentrate diet. Voles with a high incidence of glycosuria for over 6 weeks became insulin deficient. In these diabetic voles, the hepatic GKase, HKase and FDPase activities decreased considerably as a result of diminished insulin secretion and fatty degeneration of the hepatic cells. It was considered that M. arvalis would be a useful animal model in which to study disorders of glucose utilization in herbivora. PMID- 2561035 TI - Inflammation-related changes in cyclic AMP and cyclic GMP in bovine mastitis. AB - Cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) concentrations in milk and plasma samples from healthy and mastitic cows were determined by radioimmunoassay and compared with prostaglandins (PGE2, PGF2 alpha and thromboxane B2 [TXB2]), phospholipids and other relevant parameters in milk and blood. The concentrations of cAMP were about five times higher in plasma (p less than 0.01) than in milk, whereas the cGMP concentration in milk was three times higher (p less than 0.01) than that in plasma in both healthy and diseased animals. In mastitic milk, the cAMP and cGMP concentrations were 19% and 65% and in blood plasma 13% and 84% higher respectively than in healthy animals. In milk, cyclic nucleotide concentrations correlated with the markedly elevated cell count and also with the prostaglandin concentration and pH. In blood, cAMP correlated positively with phospholipids and cGMP with reduced glutathione (GSH). These changes are considered to be important in the disease process and, in particular, the increase in cGMP deserves further study. PMID- 2561036 TI - [Regression of left heart hypertrophy in hypertensive patients as a result of antihypertensive therapy]. AB - In a previous study (1) we could show a significantly more pronounced reversal of LVH with metoprolol than with gallopamil, whereas the combined therapy with atenolol and nifedipine was even more effective. We now report in 121 previously untreated hypertensive patients the longterm effect of the beta-blocker metoprolol (200 mg/die); 25 patients, mean age 43.6 yrs., follow-up 32.1 +/- 3.5 months, group A); the calcium antagonist gallopamil, 26 patients, mean age 49.7 yrs., follow-up 36.2 +/- 2.6 months, group B); the combined therapy with 50 mg atenolol and 20 mg nifedipine, 35 patients, mean age 44.5 yrs., follow-up 31.7 +/ 1.1 months, group C); 200 mg acebutolol and 20 mg nifedipine, mean age 52.1 yrs., follow-up 31.8 +/- 1.8 months, group D); 50 mg atenolol and 10 mg enalapril, mean age 43.3 yrs., follow-up 31.9 +/- 1.3 months, group E). Similar results were obtained for intraventricular septal and posterior wall thickness. Left ventricular enddiastolic dimensions remained unchanged but fractional shortenings were significantly (p less than 0.05-p less than 0.01) increased after 32 months of treatment. PMID- 2561037 TI - A genetic approach to demonstrate the role of listeriolysin O in the virulence of Listeria monocytogenes. AB - The locus of insertion of a transposon previously used to obtain a non-haemolytic avirulent mutant was identified: it is the structural gene encoding lisreriolysin O, the thiol-dependent haemolysin, now called hlyA. The gene was completely sequenced. The preliminary structural and functional study of the chromosomal region containing the gene indicates that hlyA belongs to a monocistronic transcriptional unit. If it is the case, the transposon insertion would have no major polar effect on downstream genes and would only affect hlyA expression. These results emphasize the importance of the haemolysin in the virulence of Listeria monocytogenes. PMID- 2561038 TI - Purification and characterization of cytolysins from Listeria monocytogenes serovar 4b and Listeria ivanovii. AB - Several exoproteins from Listeria monocytogenes serovar 4b (NCTC 10527) and Listeria ivanovii (ATCC) 19119, SLCC 2379), respectively, have been purified to homogeneity by thiol-disulfide exchange chromatography and gel filtration. Both strains produce a haemolytic/cytolytic protein of Mr 58 kDa, which has all the properties of a SH-activated cytolysin, the prototype of which is streptolysin O (SLO), and this protein has therefore been termed listeriolysin O (LLO). In addition a protein of Mr 24 kDa from culture supernatants of L. ivanovii co purified with LLO. The N-terminal aminoacid sequences of both proteins from L. ivanovii have been determined. By mutagenesis with transposons of Gram-positive origin (Tn916 and Tn1545), which have been introduced via conjugation into L. ivanovii, several phenotypic mutants (altered haemolysis on sheep blood agar or lecithinase-negative) were obtained. Results on the properties of these mutants will be presented. PMID- 2561039 TI - Phospholipase C in Listeria. AB - The cooperative and antagonistic effect of extracellular bacterial proteins of Streptococcus agalactiae, Rhodococcus equi, Corynebacterium ovis, and Corynebacterium haemolyticum with proteins of listerial strains of various sources, species and serovars of the membrane of sheep erythrocytes was investigated. Listeria monocytogenes and Listeria ivanovii produce, beside listeriolysin, further proteins. Their specific effect on the sheep erythrocyte membrane becomes apparent after the appearance of substances produced by the organisms under study. Ten strains of L. ivanovii produced phospholipase C responsible for the zone of incomplete haemolysis. It was inhibited by the non haemolytic sphingomyelinase D of C. ovis and C. haemolyticum. Chromatographic analysis revealed that phospholipase C splits sphingomyelin in the membrane of sheep erythrocytes. The inhibition of double haemolysis of L. ivanovii by sphyngomyelinase D of C. ovis on agar plates with washed (!) sheep erythrocytes can be utilized as a specific and rapid identification test of L. ivanovii. PMID- 2561040 TI - [Acute maternal anterior poliomyelitis in a non-endemic zone]. AB - The authors report the case of a 26 year old woman with acute anterior poliomyelitis contracted during the vaccination of her baby. Despite having been herself vaccinated in infancy she was not protected against the poliovirus. The clinical interest of this uncommon case is a severe paralytic state with definitive paraplegia. The authors suggest serologic testing of patients born before 1967 especially if they are at risk of encountering the virus. PMID- 2561041 TI - Concept of the existence of human papillomavirus (HPV) DNA in histologically normal squamous epithelium of the genital tract should be re-evaluated. AB - Because of the crucial importance of guiding current thinking in the field of HPV epidemiology, the concept of the existence of HPV DNA in histologically normal squamous epithelium was re-evaluated. A series of 102 randomly collected cervical punch biopsies, previously proved to contain the DNA of HPV types 6, 11, 16, 18, 31 or 33 by in situ hybridization were subjected to analysis for the localization of HPV DNA, i.e., whether found in the normal epithelium or at the lesion site only. This material consisted of a representative series of flat, endophytic and papillary HPV lesions with all histological grades from HPV-NCIN to HPV-CIN III, and the six HPV types in the same proportions as they occur in non-selected series of HPV lesions. Weak signals of HPV DNA were found in the basal cells in 6/102 (5.8%) of the biopsies. HPV DNA was constantly present in the parabasal cells in 25/102 (25.4%), in the intermediate cell layers in (98/102, 96%), and in the superficial cells of all 102 lesions. Noteworthy was the constant failure to reveal even weak signals of HPV DNA (of any of the six types) in histologically normal squamous epithelium of any of the 87 lesions, where such an epithelium was detectable. The present findings confirm our previous 'impression' that HPV DNA rarely if ever appears in the histologically normal squamous (or columnar) epithelium in the genital tract, when analysed using the in situ hybridization. Thus, great care should be exercised in interpreting results that suggest the discovery of HPV DNA in normal genital tract epithelium, unless based on firm documentation by in situ hybridization. PMID- 2561042 TI - Cryotherapy and CO2-laser vaporization in the treatment of cervical and vaginal human papillomavirus (HPV) infections. AB - A series of 119 women with Human papillomavirus (HPV) infections of the uterine cervix and/or vagina were included in the present study, where the efficacy of cryotherapy and CO2-laser vaporization was assessed after a mean follow-up of 14 months (SD 6 months) after treatment, as related to the natural history of the disease. Routine Papanicolaou (PAP) smears with HPV-induced changes were the basis for patient recruitment. Patients with cervical HPV lesions (HPV-NCIN, HPV CIN I or II) were randomly allocated into laser (55 women) and cryotherapy (42 women) groups. Women with combined lesions (HPV-CIN & HPV-VAIN) were treated by laser (22 patients). The cure rate after laser vaporization was practically identical to that of cryotherapy, 64% and 54%, respectively (difference not statistically significant). The success rate was significantly lower (40%) for the combined lesions (HPV-CIN & HPV-VAIN) (p less than 0.05). The residual disease encountered in patients after the first treatment with cryotherapy and laser was classified as HPV-NCIN in 78.9% and 37.0%, respectively. The number of patients is still too small to draw reliable conclusions on the effects of these therapy modes, as related to HPV type of lesion (HPV 6, 11, 16, 18, 31 and 33). The cure rates for both cryotherapy and laser in our treatment groups were significantly higher than the spontaneous regression rate (p less than 0.001), suggesting that treatment by either cryotherapy or CO2-laser vaporization significantly changes the natural history of genital HPV infections. More patients and longer follow-up are still needed, however, to fully establish the efficacy of the current treatment modalities in gynecological HPV infections. PMID- 2561043 TI - Comparison of smear specimens with biopsy specimens in a nucleic acid hybridization test for human papilloma virus (HPV) infection. AB - A total of 323 pairs of specimens from women with Papanicolaou class II or III cytology were examined for human papillomavirus (HPV) types 6, 11, 16 and 18 by spot hybridization. Each pair consisted of a representative biopsy specimen and a smear specimen from cervical, vaginal or, more rarely, vulvar lesions. We found a close correlation between HPV findings in biopsies and smears. In 83.9% (271) of the cases, both specimens were either positive for a given HPV type or negative. No discordant HPV-types in the two types of specimens were found. In 15.8% (51) of the cases, one specimen proved positive for a given HPV-type while the other specimen from the same patient was negative. In 8.0% (26) of the cases, the biopsy specimen proved positive and the corresponding smear specimen was negative. On the other hand, in 7.7% (25) of the cases we were able to detect HPV DNA in the smear specimen, whereas the corresponding biopsy specimen was negative. We suggest that a smear specimen would be advantageous for screening large groups of patients for the presence of a HPV infection in the genital tract. By using smear specimens together with biopsy specimens it is possible to maximize the number of HPV infection diagnoses. PMID- 2561044 TI - [Evaluation of crude heparin and commercial preparation of Polish production]. AB - Contents of the solid residue, ash, nitrogen, hexosamines, uronic acids, carboxyl groups, and O- and N-sulphates, as well as mean molecular weights and anticoagulant activities of the crude heparin and the commercial product were compared. The study has confirmed the literature data concerning dependence of biological activity of heparin preparations upon their molecular weight. PMID- 2561045 TI - [Effect of benzodiazepines on enzyme-metabolizing drugs. III. Activity of microsomal monooxygenases in the rat liver exposed to chlordiazepoxide and elevated environmental temperature]. AB - It has been stated that elevation of environment temperature to 28 degrees C or 35 degrees C modifies effects of chlordiazepoxide on activities of rat liver microsomal enzymic systems involved in metabolism of xenobiotics: amidopyrine, aniline and 4-nitroanisole++. PMID- 2561046 TI - [Hepatic tumors]. AB - This article show our group of liver neoplasm. We studied the presentation way in the benign and malignant tumors. Moreover we compared the therapeutics results between both groups. In our population was more frequent the malignant neoplasm. The first symptom was more frequent the malignant neoplasm. The first symptom was abdominal mass but with more general complaints in the last one. We checked the increase survival with therapeutics as ADR or CDDP. PMID- 2561047 TI - Danazol and hepatic neoplasia: a case report. AB - Although hepatic side-effects of danazol are well known, the occurrence of hepatic tumours is not well documented. We report a case of hepatocellular adenoma occurring in a 35-year-old woman with endometriosis who had been taking danazol over a three year period. PMID- 2561048 TI - Evidence against diathermy as a beneficial treatment for human papillomavirus infection of the cervix. AB - An observational study of the influence of diathermy on the rate of subsequent cytological evidence of cervical intraepithelial neoplasia and/or human papillomavirus infection among women with a histological diagnosis of cervical human papillomavirus infection is presented. After the histological diagnosis of human papillomavirus infection, 35% (23/65) of women who were not diathermied had persistence/recurrence of cytological abnormalities compared with 30% (60/203) of the women who were diathermied at the time of the biopsy. The rate ratio for further abnormality among women who did not have a diathermy relative to those who did was not significantly different at 1.25 (95% confidence interval 0.77 2.03). This study had a power of 77% to detect a true rate ratio of 2 at the 0.05 level of significance. This data does not provide strong evidence that diathermy is appropriate management for women with cervical human papillomavirus infection without evidence of dysplasia. PMID- 2561049 TI - [Antigenic structure of the foot-and-mouth virus. V. Protection of naturally susceptible animals from foot-and-mouth disease using a synthetic peptide]. AB - We have synthesized the peptide representing 135-159 VP1 sequence of A22 strain of the foot-and-mouth disease virus (FMDV). The synthetic peptide induced 100% protection of guinea pigs against the disease. Two-fold immunization of cuttle with the peptide and single immunization of sheep induced full protection of the animals against A22 strain of FMDV. PMID- 2561050 TI - Hg(II)-induced changes in DNA-circular dichroism: reversible transitions between right-handed and left-handed screwness. AB - Exposing native calf thymus DNA (in 0.1 M NaClO4, 5 mM cacodylic acid buffer, pH 6.81, 25 degrees C) to increasing concentrations of Hg(ClO4)2 produces dramatic changes in its circular dichroism (CD). Let r = [mol of added Hg(II)*/[mol DNA base]: the conservative CD spectrum of the DNA B-form, consisting of the 273 nm major (+) CD band, the 245 nm major (-) CD band, the 219 nm minor (+) CD band, and the 208 nm minor (-) CD band, becomes non-conservative in appearance at 0.01 less than r less than 0.12 and assumes the spectral characteristics of a left handed DNA double helix at 0.12 less than r greater than 1.0. The presence of a number of isoellipticity points shows that well-defined equilibria exist between the various chiroptical forms of mercurated DNA. The CD changes are totally reversible upon the removal of Hg(II), at least up to r = 1.0, demonstrating that Hg(II) keeps all base pairs in register. PMID- 2561051 TI - [Protective effect of D20 in bacteria (E. coli)]. AB - E. coli suspended in D2O showed a better survival than in H2O when the bacteria were treated with various damaging agents like UV, heat and freezing. On the contrary E. coli grown in D2O-containing media - therefore being fully deuterated - were more sensitive than normally grown bacteria. PMID- 2561052 TI - Growth inhibition of Kirkman-Robbins hepatoma by 1-(1,3-dihydroxy-2 propoxymethyl)-5,6-tetramethyleneuracil and possible mechanism of its biological activity. AB - 1-(1,3-Dihydroxy-2-propoxymethyl)-5,6-tetramethylene-uracil (DHPTU) is a newly synthesized acyclonucleoside which shows cytostatic properties. It was tested in Syrian hamster 6 days after heterotransplantation of Kirkman-Robbins hepatoma. A reduction of tumour weight by 61% was found 48 h after its intraperitoneal (i.p.) administration in doses of 20 mg per kg of body weight. Inhibition of tumour growth is accompanied by a reduction of dThd, dGuo and dTMP kinase activities in tumour cytosol (by 91% and 74% and 55%, respectively) and decrease in contents of dTTP, dGTP and dATP (by 92%, 77% and 67%, respectively) in dNTP pool. DHPTU is not phosphorylated by any tumour dN kinases, but undergoes cleavage with TU release in reaction catalyzed by the tumour cell enzyme, competitively inhibited by FA. After [14C]DHPTU or [14C]TU had been given i.p. to the animals with the tumour, 90% of the subcellular fraction labelling fell into the nuclear fraction. However, if [14C]DHPTU was administered with FA and DCF, 27% of radioisotope was found in the nuclear fractions and 68% in cytosol. Since DCF which prevented FA deamination to FB (which is not an inhibitor of the mentioned enzyme) reduces DHPTU-induced changes in activity of dN kinases and dTMP kinase in hepatoma cells, the cytostatic activity of DHPTU seems to be connected to an enzyme which releases TU from DHPTU. PMID- 2561053 TI - Characterization of simian immunodeficiency virus-specific T-cell-mediated cytotoxic response of infected rhesus macaques. AB - Four juvenile rhesus macaques were infected with simian immunodeficiency virus (SIV)MAC-Freshly isolated peripheral blood mononuclear cells (PBMC) from these SIVMAC-infected and from uninfected control macaques were assessed for cytotoxic T-lymphocyte (CTL) activity monthly for 7 consecutive months, beginning 2 months after infection. Target cells consisted of major histocompatibility complex (MHC) haploidentical parental PBMC which were stimulated with mitogen and then pulsed with heat-killed SIVMAC. CTL activity was demonstrated on all four infected animals. The effector cells are T cells which mediate cytotoxicity against SIVMAC pulsed target cells in an MHC-restricted manner. Furthermore, the cytotoxicity is virus specific and predominantly, if not exclusively, mediated by CD8+ T cells; it is also MHC class-I restricted. Incubation of target cells with leupeptin prior to the cytotoxic assay inhibited target cell generation, suggesting that viral antigens are processed via an endocytic pathway. PMID- 2561054 TI - Oligosaccharide-mediated interactions of the envelope glycoprotein gp120 of HIV-1 that are independent of CD4 recognition. AB - In this study carbohydrate-mediated interactions of the envelope glycoprotein, gp120, of HIV-1 were investigated. Oligosaccharide probes (neoglycolipids), prepared from the N-glycosidically-linked chains of the natural and recombinant forms of gp120, were used in conjunction with the intact glycoprotein to investigate reactivities with a soluble carbohydrate-binding protein (lectin) known as mannose-binding protein in human serum. Evidence is presented that the high-mannose-type oligosaccharides with seven, eight and nine mannose residues from both forms of gp120 are recognized by the serum lectin, and that these reactivities are unrelated to CD4 recognition. Reactivities of the two forms of envelope glycoprotein with macrophages derived from human blood monocytes and with the mannose-specific macrophage endocytosis receptor isolated from human placental membranes were also investigated. Evidence is presented that both forms of gp120 bind to the macrophage surface by multiple interactions in addition to CD4 binding, and that among these interactions is a carbohydrate-mediated binding to the endocytosis receptor. We propose that such carbohydrate-mediated interactions could form the basis of viral attachment to a variety of healthy and diseased tissues. PMID- 2561055 TI - A rapid method for in situ hybridization for viral DNA in brain biopsies from patients with AIDS. AB - Brain biopsy is often necessary in the diagnosis of neurological complications found in AIDS patients. We describe here a rapid method of tissue preparation and in situ DNA hybridization for detecting JC virus DNA in frozen brain biopsy sections which allows the diagnosis of progressive multifocal leukoencephalopathy to be established on the day of surgery. Once the diagnosis is established, therapeutic and management decisions can be made more easily. The commercial availability of biotinylated probes for several of the DNA viruses most frequently encountered in brain infections of AIDS patients will provide wide application of these techniques to patient management. PMID- 2561056 TI - Statistics from the World Health Organization and the Centers for Disease Control. PMID- 2561057 TI - Hairy leukoplakia, EBV: an oral history of AIDS. PMID- 2561058 TI - Deficiency of the first component of human complement. AB - C1 deficiency results from an absence or lowering of the level of one or more of the proteins C1q, C1r and C1s, which are the subcomponents of the C1 complex of the classical pathway of the serum complement system. The major clinical pattern shown in such deficiency states is an inability to deal effectively with immune complexes, resulting in the typical symptoms associated with immune-complex related diseases and a great susceptibility to recurrent bacterial infections. Both acquired and genetic deficiencies of the C1 subcomponents have been reported; the possible genetic deficiencies appear quite rare, with only 14 reports of C1q deficiency (involving 24 people) and six reports of C1r/C1s deficiency (involving 11 people) appearing in the literature to date. PMID- 2561059 TI - X-linked lymphoproliferative syndrome. AB - The X-linked lymphoproliferative (XLP) syndrome is characterized by a selective immunodeficiency to Epstein-Barr virus (EBV) manifested by severe or fatal infectious mononucleosis and acquired immunodeficiency. Prospective studies in males prior to EBV infection have demonstrate vigorous cytotoxic cellular responses, which are predominantly polyclonally activated alloreactive cytotoxic T cells. Cytotoxic T cells that recognize EBV-infected autologous B cells have been demonstrated. Fatal EBV infections in males with XLP usually result from extensive liver necrosis. Males who survive acute EBV infection demonstrate global cellular immune defects with deficient T-, B- and NK-cell responses. It is hypothesized that uncontrolled alloreactive T-cell responses triggered by EBV transformed B cells result in the immunopathy of XLP. Genetic studies have demonstrated XLP to be genetically linked to restriction fragment length polymorphisms detected with the DXS42 and DXS37 probes (from Xq26-q27). These probes make detection of carrier females and presymptomatic (EBV-seronegative) XLP males possible. Treatment of males with XLP experiencing acute EBV infection has not been successful, and current efforts are directed at prophylaxis with intravenous gammaglobulin. PMID- 2561060 TI - A DNA cassette containing a trimerized SV40 polyadenylation signal which efficiently blocks spurious plasmid-initiated transcription. AB - A head-to-tail trimer of the SV40 Bcl I-Bam H1 DNA fragment, specifying polyadenylation of RNA transcripts, was cloned as a cassette flanked by multiple restriction sites. Insertion of the trimer into several expression vectors efficiently prevented spurious expression of reporter genes resulting from transcriptional initiation in prokaryotic plasmid sequences in transfected mammalian cells. PMID- 2561061 TI - A phage-linked immunoabsorbant system for the detection of pathologically relevant antigens. AB - This report describes a novel system for the immunological detection of immobilized antigen. The detection of herpes simplex virus (HSV) antigen was used as an example. Bacteriophage M13, containing the E. coli lac Z gene, was used as the "reporter" molecule in an immunoassay which is otherwise analogous to the enzyme-linked immunoabsorbant assay (ELISA). Briefly, HSV infected cells were incubated with a mouse monoclonal antibody specific for HSV antigen, followed by rabbit anti-mouse serum and mouse anti-M13 serum. Immune complexes were incubated with viable M13 phage. M13 binding was due to the presence of M13 antibodies, whose presence ultimately depended on the binding of monoclonal antibody to HSV. Phage was recovered by elution in pH = 11. Recovered phage was used to infect E. coli. M13 was quantitated by either plaque assay or by an assay for phage-induced beta-galactosidase activity in appropriate E. coli strains. The amount of M13 recovered was proportional to the number of HSV infected cells probed. Therefore, M13 served as a "bio-amplifiable tag" to antibody, as enzymes do in the ELISA. Since M13 is viable, its signal can be amplified by infection of susceptible bacteria, and the promise for an enormously sensitive immunoassay exists. The sensitivity of the assay described here is compared to the ELISA in the detection of HSV infection cells, as an example of the novel assay's potential. Significantly, the novel assay was more sensitive than the ELISA when samples were tested under identical circumstances. This technique is called the phage linked immunoabsorbant assay (PHALISA), by analogy to the ELISA. PMID- 2561062 TI - Monoclonal antibody purification: choice of method and assessment of purity and yield. AB - In this article we discuss our choices of monoclonal antibody separation methods for the applications which face us most frequently. We explain the rationale behind these choices, to help other users make their own choices. The review is intended to be brief and selective; references to detailed reviews are provided. PMID- 2561063 TI - Construction of cloning/sequencing vectors with an alternative polylinker. AB - This paper describes the construction of a new plasmid and M13 phage cloning vector in which the 54-bp polylinkers of pUC19 and of M13mp8 are replaced with a 45-bp chemically synthesized polylinker with different restriction sites. The new polylinker is inserted in frame at the N-terminal end of the beta galactosidase lac Z fragment. The plasmid was named pLH1, the phage M13LH1. PMID- 2561064 TI - High resolution imaging of receptor binding in analyzing neuropsychiatric diseases. AB - A method for analyzing high resolution imaging of receptor binding activity in human post-mortem brain specimens is described. The autoradiography technique employed is based on methods previously described by others in which coverslips dipped in tritium-sensitive nuclear track emulsion are placed over a tissue section that has been incubated in a medium containing radioactively-tagged ligand. With this approach, there is a 370-fold increase in resolution from approximately 120 microns available with tritium-sensitive films to 0.33 micron attainable with the emulsion approach. Since the coverslip autoradiogram remains superimposed on the tissue section, individual grains can be routinely quantitated in specific cell types and discrete subregions of the neuropil with the aid of a user-interactive image processing system. Overall, the improved resolution that this approach provides makes it possible to determine whether a particular neuronal sub-type may be preferentially altered by disease processes affecting the brain. PMID- 2561065 TI - A simple and versatile method for the preparation of vector-primers by adapter end-primer ligation. AB - A group of efficient cDNA cloning strategies employs vector-primers where cDNA synthesis starts from the oligo(dT)-primer tail, which is conventionally attached to cloning vectors by use of terminal deoxynucleotidyl transferase. An alternative, efficient and more versatile method of vector-primer preparation is to directly ligate, by use of T4 DNA ligase, a double-digested vector, e.g., pTZ18R/Pst I/Bam HI, to a synthetic (Bam HI)-adapter-end-primer, 5'-pGATCC-Tn or 5'-pGATCC-site-specific sequence. The use of a utility-vector containing a sizable spacer between the two selected restriction sites enables unambiguous separation on agarose gels of the double-digested vector precursors from single digested ones, further simplifying the vector preparation. The adapter-end-primer ligation method can be applied to any suitable vectors with multiple cloning sites for the preparation of not only oligo(dT)-tailed, but also site-specific sequence-tailed vectors. Thus, the method enables the cDNA cloning of total poly (A+)-mRNAs, as well as specific RNA or mRNA species with or without poly(A)-tail. PMID- 2561066 TI - The guanine nucleotide regulatory protein-coupled receptors for nucleosides, nucleotides, amino acids and amine neurotransmitters. PMID- 2561067 TI - The erb-A family receptors for thyroid hormones, steroids, vitamin D and retinoic acid: characteristics and modulation. PMID- 2561068 TI - Cell surface receptors that serve to transport ligands. PMID- 2561069 TI - Regulation of cell excitability and solute transport. PMID- 2561070 TI - Growth factors and their receptors. PMID- 2561071 TI - [Heart insufficiency. Physiology and physiopathology of muscle contraction of the heart]. AB - To understand physiopathology of contractibility in heart failure is needed the previous knowledge of normal muscle contractibility, calcium regulation, methods of assessment of isolated cardiac muscle and in situ heart contraction. Models of calcium movements during rest and contraction are presented and concepts of preload, afterload, Vmax and Po introduced. The importance of assessment of cardiac diastolic function is referred. Biochemical alterations in the failing myocardium are presented. PMID- 2561072 TI - A subsequence-specific DNA-binding domain resides in the 13 kDa amino terminus of the bacteriophage Mu transposase protein. AB - We have previously reported that the 13 kDa amino terminus of the 70 kDa bacteriophage D108 transposase protein (A gene product) contains a two-component, sequence-specific DNA-binding domain which specifically binds to the related bacteriophage Mu's right end (attR) in vitro. To extend these studies, we examined the ability of the 13 kDa amino terminus of the Mu transposase protein to bind specifically to Mu attR in crude extracts. Here we report that the Mu transposase protein also contains a Mu attR specific DNA-binding domain, located in a putative alpha-helix-turn-alpha-helix region, in the amino terminal 13 kDa portion of the 70 kDa transposase protein as part of a 23 kDa fusion protein with beta-lactamase. We purified for this attR-specific DNA-binding activity and ultimately obtained a single polypeptide of the predicted molecular weight for the A'--'bla fusion protein. We found that the pure protein bound to the Mu attR site in a different manner compared with the entire Mu transposase protein as determined by DNase I-footprinting. Our results may suggest the presence of a potential primordial DNA-binding site (5'-PuCGAAA-3') located several times within attR, at the ends of Mu and D108 DNA, and at the extremities of other prokaryotic class II elements that catalyze 5 base pair duplications at the site of element insertion. The dissection of the functional domains of the related phage Mu and D108 transposase proteins will provide clues to the mechanisms and evolution of DNA transposition as a mode of mobile genetic element propagation. PMID- 2561073 TI - Susceptibility of Salmonella typhimurium and Salmonella typhi to oxygen metabolites. AB - The susceptibility of Salmonella typhimurium LT2 and S. typhi 1079 to oxygen metabolites were compared. S. typhimurium LT2 and S. typhi 1079 were killed to an equal extent (about 40%) by the xanthine-xanthine oxidase (200 mU/ml) system. Among the various scavengers of oxygen metabolites, catalase alone inhibited the killing of S. typhimurium LT2 and S. typhi 1079 by the xanthine-xanthine oxidase system, indicating that hydrogen peroxide contributed to the killing of Salmonellae. The respiratory burst of murine macrophages was efficiently triggered by the ingestion of S. typhimurium LT2, S. typhimurium SL1102, and S. typhi 1079 and all to the same extent. However, in the range of the concentration of hydrogen peroxide produced by murine macrophages, neither S. typhimurium LT2 nor S. typhi 1079 were killed. Only S. typhimurium SL1102, a rough mutant of S. typhimurium LT2, was markedly susceptible under these conditions. The findings suggest that both S. typhimurium LT2 and S. typhi 1079 are resistant to oxygen dependent killing mechanisms. PMID- 2561074 TI - [Glomus jugulare tumor. Apropos of 2 cases]. AB - We present two clinical cases of jugular glomus in which different kinds of treatment have been adopted. In literature we can find a review of the utility and morbidity of the different methods of exploration as well as the results obtained by the different authors according to the kind of treatment. Nevertheless we don't come to definite conclusions due to the fact that the classifications and curing criteria very in the different series. PMID- 2561075 TI - [Malignant tumor of the external auditory canal]. AB - The neoplasms from the modified apocrine glands of the external auditory canal are usually included under the term ceruminoma without any reference to their histology, clinical course or prognosis. It seems better to classify them according to their histologic pattern in the following types: 1.--Ceruminous adenoma, 2.--Adenoid-cystic carcinoma, 3.--Adenocarcinoma, and 4.--Pleomorphic adenoma (mixed tumour). We describe a case of adenoid-cystic carcinoma originated at postero-superior wall of the external auditory canal which presented as an otitis of long duration, and was treated surgically and received post-operative radiotherapy. The patient is at present free of disease. We review the literature on this group of tumours of the EAC. PMID- 2561076 TI - [Image diagnosis and pleomorphic adenoma]. AB - We examined the imaging technical findings of 11 benign pleomorphic adenomas of major salivary glands. The imaging technical included sialography, echography, computed tomography and gammagraphy. We compared the diagnostic usefulness of each of these imaging technical. The purpose of this paper is to identify the actual advantages, disadvantages and uses of these diagnostic methods. PMID- 2561077 TI - The measles dilemma: what about it? PMID- 2561078 TI - Low field magnetic resonance imaging of focal hepatic masses at 0.02 T. AB - The diagnostic utility of extremely low field magnetic resonance (MR) imaging was evaluated in 25 patients with focal hepatic masses, including 17 with primary (n = 7) or secondary (n = 10) malignant neoplasms and 8 with benign lesions (6 hemangiomas). The findings were compared with the results of computed tomography (CT). Out of 16 patients with malignant tumors demonstrated by both modalities, the diagnostic information from MR imaging was equal to or better than that from CT in 6 patients and inferior to CT in 10. Shortcomings of MR were mainly due to low signal-to-noise ratio and poor spatial resolution, resulting in an image quality inferior to that obtained at higher field strengths. Considering these facts, together with the long imaging times required, low field MR cannot be recommended for general use in the evaluation of hepatic masses. On the other hand, our results indicate that this technique may be useful in establishing the diagnosis of hepatic hemangioma. PMID- 2561079 TI - Treatment of liver metastases by arterial injection of adriamycin/mitomycin C lipiodol suspension. AB - Sixty-one patients with liver metastases were treated with hepatic arterial injection of Adriamycin/Mitomycin C oil suspension (ADMOS). The liver metastases originated from the gastrointestinal tract in 41 patients and from other organs in 20 patients. Sixty-nine liver tumors were analyzed in these 61 patients. Computed tomography (CT) after ADMOS injection (Lip-CT) gave more information in 19 cases (31%) than enhanced CT with a water-soluble contrast medium. With Lip CT, Lipiodol tumor enhancement was observed in 64 of 69 lesions. The patterns of Lipiodol uptake in the tumors were classified into 4 types: Homogeneous accumulation (20%), heterogeneous accumulation (16%); accumulation with a central defect (57%); and no accumulation (7%). A tumor response was achieved in 41 of 69 lesions (59%). Anticancer effects were also shown as a decrease in serum CEA levels. The one year survival rate estimated by the Kaplan-Meier method was 43 per cent and the median survival time was 337 days. The results were better among patients who received multiple doses of ADMOS. Only minimal side effects were associated with ADMOS. The method is considered to be an excellent diagnostic and therapeutic procedure for liver metastases. PMID- 2561080 TI - [The effect of cobalt on the dynamics of membrane-bound calcium in the cerebral cortex structures]. AB - Cobalt (Co2+) influence on calcium bound (Cab2+) dynamics detected by a fluorescent chlorotetracyclin probe (CTC), as well as bioelectric activity were studied in the feline vital brain cortex preparation with contact microscopy and microelectrode technique. It has been found that Co2+ applied on the preparation surface or injected ionophoretically into microzones under microscopy produced a distinct decrease of CTC-Cab2+ fluorescence intensity. This indicates a decrease of Cab2+ content in the hydrophobic membrane loci. The effect is reversible and depends on Co2+ concentration. Neuronal excitation appeared after the fluorescence decrease up to 5-12% to the start level. It has been concluded that Cab2+ content dynamics plays an important role in the Co2+ "seizure" discharges mechanisms in the cortex. PMID- 2561081 TI - [Detection of new metabolic paramagnetic centers in the lung tissue by ESR method]. AB - An ESR signal with g = 2.0025 and delta H = 0.2 mT was recorded at 77 K in lung tissues of guinea-pigs, mouse and rats. The signal intensity changed in the lung tissues of animals exposed to some chemicals. A relationship between new paramagnetic centres and the lung surfactant system is suggested to exist. PMID- 2561082 TI - [Segmental flexibility of Fc-region of spin labeled 2 subclasses of IgG molecule from cow blood and local conformational mobility of their carbohydrate components]. AB - Spin-label was bound to carbohydrates of Fc-region of two subclasses of the IgG molecule from the blood serum of healthy and ill cows. By the value of rotational correlation time (10 nsec) internal flexibility of Fc-region of both IgG subclasses and by value of the order parameter (S = 0.9)--rigid attachment of oligosaccharide chains to protein parts of the molecules were found. At the same time the states of IgG1 an IgG2 molecules determined by dynamic steric characteristics for ill and healthy cows did not differ. PMID- 2561083 TI - [Comparison of DNA interaction with sphingomyelin, spermine and magnesium ions using a spin label method]. AB - Identical interactions of liposomes from sphingomyelin, spermine and magnesium ions with DNA was shown by spin probe method using spin-labeled 9-aminoacridine. The interactions proceed in the phosphate groups at the expense of hydrophobic part of phospholipid. The functional role of sphingomyelin--DNA interaction in matrix biosyntheses is discussed. PMID- 2561084 TI - [Study of erythrocyte dehydration using spin labels]. AB - Possibility of studying erythrocyte dehydration by ESR-spin probe is substantiated. Dehydration of erythrocytes in relation to osmolarity of sodium chloride solutions is investigated. The results are shown to agree with the data obtained by radioisotope method. PMID- 2561085 TI - Farnesylacetone, a sesquiterpenic hormone of Crustacea, inhibits electron transport in isolated rat liver mitochondria. AB - Farnesylacetone (C18 H30 0) is a male hormone extracted from the androgenic gland of crab, Carcinus maenas. Appropriate enzymatic assays, as well as spectrophotometric studies, indicate that micromolar concentrations of farnesylacetone interact with the electron transport pathway of rat liver mitochondria. By the use of artificial electron donors and electron acceptors, it is shown that farnesylacetone immediately inhibits the electron transfer within complex I (NADH ubiquinone reductase activity) and complex II (succinate ubiquinone reductase activity). It is proposed that farneylacetone could interact with these two complexes of the respiratory chain at the level of the iron-sulfur centers implicated in the dehydrogenase activities. These observations are compared with the results obtained with terpenic molecules which interact with mitochondrial respiration. PMID- 2561086 TI - [Effect of scavengers of superoxide radicals in reducing ischemic injury of skin flaps]. AB - In recent years, an extensive attention has been paid to the effects of oxygen free radicals on tissue injury. Although some studies have been demonstrated that the oxygen free radicals are implicated in the pathogenesis of ischemic injury in island skin flaps, none of the experimental study has been domestically reported. The authors used the island flap of the rats as an animal model to investigate the effectiveness of free radical scavengers in preventing reperfusion injury after ischemia in island skin flap. The results suggest that the scavenger of superoxide radical--superoxide dismutase (SOD) which was directly injected into the flap before and after reperfusion increases the survival rate of the flap obviously (93.6% v.s. control 63.7%). This findings are consistent with the theory that the ischemic tissue injury is caused by the oxygen free radicals produced at the time of tissue reperfusion. PMID- 2561087 TI - [The role of selective bronchial arterial infusion in the comprehensive treatment of advanced bronchial carcinoma]. AB - Bronchial arterial infusion (BAI) of cisplatinum and procaine was performed in 20 patients with confirmed, advanced bronchogenic cancer. The short-term response to BAI and survival period were reported. The responsive rates of CR and PR are 65% (75% in small cell carcinoma and 65% in squamous cell carcinoma). The overall mean survival period is 11.5 months (8 months in small cell carcinoma and 12 months in squamous cell carcinoma). We would like to point out that the comprehensive treatment based on BAI should be emphasized in order to improve the survival period. BAI followed by radiation therapy have an effect on completing each other in the therapy of squamous cell carcinoma. But, BAI should take the second place in treatment of small cell carcinoma. PMID- 2561088 TI - The hypotensive effect and antifungal activity of 3,3'-dihydroxy-alpha,beta diethyldiphenylethane. AB - 3,3'-Dihydroxy-alpha,beta-diethyldiphenylethane (I), an isomer of hexestrol (II), was found to have a strong hypotensive effect on rats and antifungal activity against some pathogenic fungi. The hypotensive effect of I (-100.0 +/- 21.0 mmHg, 10 mg/kg, i.v.) was stronger than that of II (-40.0 +/- 2.60 mmHg, i.v.). Unlike II and other oxystilbene-related compounds already tested, I showed prolonged hypotensive action at the dose of 10 mg/kg, and the blood pressure was not restored to the original level within 20 min. I also showed antifungal activity against all fungi tested (minimal inhibitory concentration: 10-120 micrograms/ml). It should be emphasized that by shifting the phenolic hydroxyl group to m-position, the biological activities were greatly increased. PMID- 2561089 TI - Infection and cross-infection in a paediatric gastro-enteritis unit. AB - A two month study to investigate the incidence of nosocomial infection was conducted in a paediatric gastroenteritis ward of a black academic hospital. Enteric pathogens were identified on admission in 61 (47.2%) of 129 patients; 56 bacterial and 25 viral. Six per cent of patients had a combination of bacterial and viral pathogens. Enteric pathogens most frequently identified on admission were Campylobacter jejuni in 22%. Rotavirus in 19.3%, EPEC in 10.8% and Shigella spp. in 6.9% patients. Twenty six (20%) patients had more than 1 enteric pathogen. The nosocomial infection rate was recorded at 17.1%. EPEC occurred most commonly in 5.3% patients, Salmonella typhimurium in 4.6% and Shigella spp. in 2.3%. Nosocomial infections increased the mean length of hospital stay from 7.2 20.2 days. Contributory factors to the spread of nosocomial infection were the unsatisfactory methods of bathing patients and giving naso-gastric feeds. PMID- 2561090 TI - Biochemical evaluation of organ cultures from primate flexor tendons. AB - Recently our laboratory has reported, in a lacerated flexor tendon model, that the "early turnover" phase of the repair process extends for a longer period of time in vivo than previously documented. The extensive turnover of the collagenous matrix was consistent with the presence of collagenolytic activity in repairing tendon tissue and suggested a possible regulatory role for neutral metalloproteinases in flexor tendon repair. However, these in vivo observations could not distinguish the relative contribution by the tendon fibroblasts from that of the surrounding sheath and vascular tissue elements. To further define these interrelationships, the present study investigates the repair process of the flexor tendon in an in vitro tissue culture environment. The sequential changes in matrix formation were defined (i.e., proteoglycans/glycosaminoglycans, glycoproteins, and collagenous proteins). The concomitant production of neutral metalloproteinases as well as prostaglandin E2 was determined in relation to net tissue repair. Profundus flexor tendon segments were obtained from young adult Macaca nemestrina monkeys and maintained in organ culture for periods from 4 days through 9 weeks. Initially (at 2 wks) there was an increase in both sulfated and nonsulfated glycosaminoglycans, which preceded the onset of maximal collagen protein formation. By 6 and 9 weeks of in vitro repair, of the lacerated tendon segments, there was a significant increase in net collagen formation. Neutral metalloproteinase activity increased early in the repair period, from the 4th to 9th day, and decreased thereafter through the 9th week of culture. Functionally the enzyme appeared to be a gelatinase. The temporal pattern of in vitro collagen synthesis in relation to the gelatinase activity support the hypothesis that regulation of this enzyme(s) may be a critical factor in mediating the flexor tendon response to injury. PMID- 2561091 TI - Hypertrophic chondrocytes produce immunoreactive collagenase in vivo. AB - A monospecific, polyclonal antibody to neutral collagenase purified from rat myometrial cells was used to examine decalcified sections of normal and rachitic rat long bones, including epiphyses, for presence of immunoreactive collagenase in situ. Reactive antigen was uniformly present in hypertrophic chondrocytes of articular and epiphyseal plate cartilage. Preincubation of antibody with excess homogeneously purified antigen blocked the staining, indicating specificity of the staining for the antigen. Reaction was present in the borders of the enlarging lacunae, suggesting functional importance of this enzyme in the process of lacunar enlargement prior to mineralization. No detectable enzyme was observed in osteoblasts, osteocytes, or osteoclasts. No difference between expression of collagenase by bone cells in rachitic or normal bone was seen. Thus, at the level of sensitivity of this procedure, neutral collagenase appears to function in mature rat bone only in enlarging chondrocyte lacunae. PMID- 2561093 TI - [Experimental research on the pathogenic mechanisms in adult respiratory distress syndrome]. AB - In order to study the pathogenic mechanisms of adult respiratory distress syndrome (ARDS), the neutrophil aggregation activity, platelet aggregation rate and levels of thromboxane, prostacyclin, superoxide dismutase, lipoperoxides, neutrophil elastase, alpha 1-antitrypsin and angiotensin conversion enzyme were measured in 84 experimental dogs. Under the action of activated complement C5a, polymorphonuclear leukocyte aggregation with increased release of oxygen radicals play an important role in the pathogenesis of ARDS. PMID- 2561092 TI - [Treatment of gestational trophoblastic neoplasms with methotrexate and citrovorum factor rescue: analysis of 43 cases]. AB - From Jan. 1979 to Nov. 1987, 38 patients with invasive mole and 5 patients of choriocarcinoma were primarily treated with methotrexate and citrovorum factor (MTX-CF) rescue. Thirty-two patients had non-metastatic disease (stage I) and 11 had metastatic disease (stage IIA in 5 cases, stage IIB in 3 cases and stage IIIA in 3 cases). Complete remission was achieved in 28 (87.5%) of 32 patients with non-metastatic disease and in 9 (81.8%) of 11 patients with metastatic disease. Six patients with MTX-CF resistant tumors subsequently achieved complete remission with intravenous infusion of KSM and/or AT 1258. All patients were followed up periodically, 22 of them have been followed up for over 2 years, the longest duration of follow-up being 7 years. Seven of the 14 patients with preserved uterus became pregnant after recovery. All children grew up normally. PMID- 2561094 TI - [Estimation of intracellular free calcium concentration in patients undergoing digitalis treatment and in patients presenting digitalis toxicity]. AB - Delayed afterdepolarizations occur under conditions in which there are large increase in the intracellular Ca and may be considered as one of the important mechanisms for digitalis-induced arrhythmias. Intraplatelet free calcium (Ca2i+) was measured in 25 patients, using the fluorescent indicator Quin-2. The patients included 15 with glycosides treatment (Group A) and 10 with arrhythmias indicating the presence of digitalis intoxication (Group B). The average Ca2i+ level in Group A was higher than the normal value but did not reach a statistical significance (170.8 +/- 9.09 vs 161.6 +/- 30.2, NS). The Ca2i+ level during digitalis toxicity in Group B was significantly higher than that in Group A (201.7 +/- 17.65 vs 170.18 +/- 91.09, P less than 0.01) as well as the normal value (P less than 0.01). With the disappearance of digitalis toxicity the Ca2i+ level was significantly decreased from 201.7 +/- 17.65 to 171.4 +/- 14.08 (P less than 0.001). Following the elimination of digitalis toxicity the increased digoxin concentration also declined. However, the decline of digoxin concentration did not correlate well with the decline of Ca2i+ (r = -0.57, P greater than 0.05). There is a close association between digitalis-induced arrhythmias and the change of Ca2i+ level, compatible with the tentative mechanism of delayed after depolarization (triggered activity). The poor correlation between the Ca2i+ level and digoxin concentration during the process of digitalis toxicity may suggest the presence of factors other than glycosides concentration to cause increased Cai2+ and arrhythmias. PMID- 2561095 TI - [Ectopic ACTH syndrome caused by bronchial carcinoid]. AB - A 35-year male patient with clinical and biochemical manifestations of severe hypercorticism was thought to have Cushing's syndrome of pituitary origin. However, the surgically removed pituitary adenoma was not confirmed by pathological examination. There was no improvement after transsphenoidal microsurgery Chest CT scan showed a small mass located at the upper-lobe of the right lung. This tumor (1.8 x 1.0 x 1.0 cm) was removed and one course of radiotherapy with linear accelerator was given. Remission was achieved clinically and biochemically after these therapies. The diagnosis of bronchial carcinoid was confirmed by pathological findings. Metastasis of a lymph node was also proved. The tumor cells was found to contain ACTH and related peptides with radioimmunoassay, immunocytochemistry and electron microscopy. Gel filtration of the tumor extracts showed molecular heterogeniety of ACTH related peptides and that the big-form were in large percentage. PMID- 2561096 TI - [The glucose tolerance test and insulin secretion test as risk factors in liver cancer surgery]. AB - To investigate the predictive value of oral glucose tolerance test (O-GTT) and insulin secretion test (IST) on the risk of hepatectomy in liver cancer patients, we through double-blind method, compared the results of these two tests, clinical course of the patients, and the pathological findings. It was found that: 1) The positive prediction value, negative prediction value, and accuracy of O-GTT were 79.2%, 94.4%, and 85.7%, the corresponding figures of IST were 55.6%, 100%, and 61.9%, respectively. 2) Pattern of the curve of O-GTT believed to depend on roughly normal hepatic energy metabolism and islet secretion capacity suggested better tolerance for hepatectomy. 3) A part of the patients with advanced HCC had a depressed islet secretion capacity. 4) The delta IST/delta O-GTT showed an accurate negative prediction for hepatectomy when the ratio was less than 50 x 10(-9). 5) Apart from O-GTT and delta IST/delta O-GTT, the severity of the hepatitis and cirrhosis should be taken into account in the decision of carrying out hepatectomy. PMID- 2561097 TI - Reaction of trans-2-chlorovinylarsine oxide with polydeoxynucleotides. AB - Trans-2-chlorovinylarsine oxide (in DCl/acetone-d6) was added to various polydeoxynucleotides. The arsenical did react with poly[dG].poly[dC], releasing guanine, and resulting in a partial apurinic duplex. PMID- 2561098 TI - Excision of residual masses after platinum based chemotherapy for non seminomatous germ cell tumours. AB - Between January 1980 and June 1987, 42 patients receiving platinum based combination chemotherapy for advanced non-seminomatous germ cell tumours had residual masses, detected by computed tomography, after four or six treatment courses without tumour marker evidence of active disease. Resection of retroperitoneal (n = 32), pulmonary (n = 4) or thoracoabdominal (n = 2) disease revealed residual malignancy in nine patients (21%), differentiated teratoma in 14 (33%) and fibrosis or necrosis in 15 (36%). Laparotomy showed no evidence of a mass in four instances. Of the 42 patients, 14 had malignant teratoma undifferentiated in the primary tumour only one of whom (7%) had evidence of malignancy in the specimen resected post-chemotherapy. Conversely, six of 15 patients (40%) whose primary tumour was malignant teratoma intermediate had residual malignant tissue after treatment. With a median follow up of 36 months from post-chemotherapy surgery, 36 patients (86%) are continuously disease-free. Relapses occurred in one of nine patients with residual malignancy (11%), three of 14 with differentiated teratoma (21%), one of 15 with necrosis or fibrosis (7%) and in one patient who had a normal laparotomy. Four patients have died from their tumours, but two are currently disease-free following further surgery and chemotherapy for relapse. Neither primary nor post-chemotherapy histology was predictive of relapse, and although relapse was numerically more common in patients whose residual mass was incompletely excised (three of 12), this was not statistically significant. PMID- 2561099 TI - Chemoprotection by 9-aminoacridine derivatives against the cytotoxicity of topoisomerase II-directed drugs. AB - The effect of three acridine derivatives, 9-aminoacridine (9AA), 4'-(9 acridinylamino)-methanesulphon-O-anisidide (O-AMSA) and quinacrine were compared in their ability to protect against the cytotoxicity of amsacrine, 9-[[2-methoxy 4-[(methylsulfonyl)amino]phenyl]amino)-N,5-dimethyl-4- acridine-carboxamide (CI 921), N-[2-(dimethylamino)ethyl]acridine-4-carboxamide (AC), etoposide, mitoxantrone and doxorubicin. Cytotoxicity was measured in vitro by clonogenic survival assay and in vivo by life extension assays. All three acridine derivatives protected a Lewis lung cell line in vitro against CI-921, with 9AA having the highest activity. Cellular uptake of [14C] CI-921 by cultured Lewis lung cells was unaffected by 9AA, and slightly stimulated by O-AMSA and quinacrine. 9AA protected Lewis lung cells in vitro against the cytotoxicity of amsacrine, CI-921, AC and etoposide, partially against mitoxantrone but not against doxorubicin. A similar result was obtained with the human melanoma cell line MM96, where 9AA protected against CI-921 but not against doxorubicin toxicity. 9AA protected P388 leukaemia in vivo against amsacrine, CI-921 and AC cytotoxicity, partially against etoposide but not against mitoxantrone or doxorubicin. 9AA also protected against animal toxicity caused by high dose amsacrine and partially against CI-921 toxicity. It is hypothesized that DNA intercalating chemoprotectors act by restricting the conformational flexibility of the DNA and thus the ability of topoisomerase II to form a 'cleavable complex' in which the DNA is covalently linked to the enzyme. PMID- 2561100 TI - A phase II trial of TCNU in patients with squamous, adeno and large cell carcinoma of the lung. PMID- 2561101 TI - Age-related changes in DNA polymerase alpha expression. AB - DNA polymerase alpha isozymes differing in specific activity and affinity of binding to DNA were purified from human fibroblasts derived from donors of different ages. Fetal-derived fibroblasts expressed a single, high-activity enzyme (A2), with high affinity of binding to DNA. Adult-derived fibroblasts exhibited two forms of DNA polymerase alpha, one identical to the fetal enzyme, and a second with about tenfold less activity showing low affinity of binding to DNA (A1). The ratio of DNA polymerase A2/A1 decreased dramatically with age, from 100% A2 in fetal-derived fibroblasts to about 94% A1 in fibroblasts derived from a 66-year-old donor. The DNA binding affinity of polymerase alpha A1 from adult derived fibroblasts increased concomitant with a significant increase in activity when the enzyme was treated with phosphatidylinositol-4-monophosphate (PIP), or with inositol-1, 4-bisphosphate (I(1,4)P2). The enzyme reverted back to a less active form, with loss of the noncovalently bound I(1,4)P2, as a function of time. When permeabilized human fibroblasts with low DNA excision repair capacity were treated with 7,8-dihydrodiol-9,10-epoxybenzo(a)-pyrene (BPDE) in the presence of 32P-ATP, phosphatidylinositol, and cycloheximide, excision repair was initiated and 32P-labeled DNA polymerase alpha was recovered in the absence of de novo protein synthesis. DNA synthesis associated with either scheduled DNA synthesis or BPDE-initiated excision repair declined as a function of increased age in human cells. The data suggest that the decline in both DNA excision repair associated and mitogen-activated DNA synthesis may be correlated with decreased total intracellular levels of DNA polymerase and with the decline in polymerase alpha activity as a function of age, that DNA repair-associated initiation of DNA synthesis in adult-derived cells may increase with activation of a pool of low activity DNA polymerase alpha, and that DNA polymerase alpha activity increases as a function of enzyme interaction with a component of the PI phosphorylation cascade. PMID- 2561102 TI - Altered intracellular protein degradation in aging: a possible cause of proliferative arrest. AB - Many proteins that control cell-cycle progression are short-lived. Therefore, alterations in protein degradation are as likely as changes in transcription and/or translation in causing the proliferation arrest of senescent cells. Several different pathways of intracellular protein degradation have been identified, and both cytosolic and lysosomal pathways operate in most cells. We have used red cell-mediated microinjection to study degradation of radiolabelled proteins introduced into IMR-90 human diploid fibroblasts at early and late population doubling levels. Lysosomal pathways of protein degradation are reduced in senescent cells, and this defect may account for many characteristics of aging, including the accumulation of posttranslationally altered proteins. These abnormal proteins may then stimulate cytosolic, ubiquitin-dependent proteolytic pathways that are also responsible for the degradation of crucial regulatory proteins. Unknown short-lived proteins are also required for some step in lysosomal proteolysis, and this connection between the two degradative systems may cause the age-related changes in protein degradation to be progressive. Several experimental approaches are available to test whether altered protein degradation significantly contributes to proliferative arrest of senescent cells. PMID- 2561103 TI - Levels of DNA methylation in diploid and SV40 transformed human fibroblasts. AB - Total 5-methylcytosine (5mC) has been measured in the DNA of cultured human cells by HPLC chromatography. In agreement with other reports, the level of 5mC declines continuously during the serial passaging of diploid fibroblasts, strain MRC-5. On the contrary, fibroblasts infected with SV40, which have acquired most of the properties of transformed cells, maintain a constant level of 5mC, prior to dying out in the crisis period. MRC-5 and Werner's syndrome fibroblasts fully transformed and immortalised by SV40, also maintain a constant level of 5mC. These results indicate that the ability of cells to maintain DNA methylation is a necessary but not a sufficient step for immortalisation. PMID- 2561104 TI - Replicative senescence and differentiated gene expression in cultured adrenocortical cells. AB - We have used a differentiated endocrine cell type, the adrenocortical cell, to investigate the interrelationship of senescence and differentiation, the effects of the environment on differentiated gene expression, and the interrelationship of differentiated gene expression and proliferation. In bovine adrenocortical cells, expression of some differentiated functions is maintained to very late points in the replicative life span, whereas expression of others is lost at various times prior to senescence. There is clonal variation in the rate and extent of loss of functions. For steroid 17 alpha-hydroxylase, in situ hybridization shows that the observed decline in induction of 17 alpha hydroxylase mRNA during senescence results from a decline in the fraction of cells expression the gene. Descendants of expressing cells in the primary cell population randomly become nonexpressing. Among clones there is a correlation between the fraction of cells expressing the gene and remaining replicative potential, although several experiments show no direct mechanism linking replicative senescence and 17 alpha-hydroxylase expression. Transfection with SV40 early region genes also dissociates the decline in growth and the change in 17 alpha-hydroxylase expression. SV40 T antigen selectively affects growth; expression of 17 alpha-hydroxylase is stabilized either in the on state, when cells are transfected early in the culture life span, or in the off state, when senescent cells are transfected. Thus, although the switching off of 17 alpha hydroxylase expression and the loss of replicative potential are independent events, the switching process requires DNA replication. Because the switch is irreversible, changes in replicative potential occurring after the switch-off event do not affect the state of expression of the switched-off gene. Changes in differentiated cell properties and changes in replicative potential may be two facets of a general phenomenon of stochastic changes in gene expression in normal cells during senescence. PMID- 2561105 TI - Collagenase production by early and late passage cultures of human fibroblasts. AB - Comparison of the proteins secreted by early and late passage cell cultures of human fibroblasts revealed a high level of immunoreactive collagenase (Mr = 55,000 Da and 58,000 Da) in the late passage cell culture conditioned medium. Both molecular weight species reacted with a monoclonal anticollagenase antibody and were apparently glycosylation varaents of the same protein. The question of whether the apparent age-dependent differences in collagenase synthesis reflected changes in protein synthesis or secretion was addressed by assaying immunoreactive collagenase and collagenase mRNA. Immunofluorescence microscopy of cellular collagenase revealed that the percentage of collagenase positive cells ranged from 1 to 6% (early passage) to 35 to 46% (late passage) indicating that the late passage cells had higher basal levels of collagenase synthesis. Later passage cultures also secreted higher levels of immunoprecipitable collagenase into the culture medium and Northern analysis established that the basal level of collagenase mRNA was also 10 times greater in late passage cells. High basal levels of collagenase were also observed in fibroblasts cultured from an in vivo aged donor and from donors with Werner's syndrome. Collagenase production was induced in both early and late passage cell cultures by exposure to fibroblast extracellular matrix, fibroblast conditioned media, polypeptide growth factors, or phorbol esters. The induced levels were always greater in the late passage cell cultures than in the early passage cell cultures. PMID- 2561106 TI - Ulcerative colitis in remission: it is possible to predict the risk of relapse? AB - A series of anamnestic, clinical, endoscopic and histologic parameters was evaluated in 64 patients with ulcerative colitis in remission to assess whether any of these had predictive value of relapse. All patients with quiescent disease were followed for at least 6 months. The statistical analysis was carried out by means of likelihood chi 2 test, receiver operating characteristic curve, Youden index and stepwise logistic regression. Eighteen patients out of 64 (28%) had a relapse, in a mean period of 9 months after the beginning of the study. The findings of our study suggest that only three factors were useful in selecting a subpopulation with a higher risk of recurrence: (a) a fiber-poor diet; (b) a number of previous episodes of relapse higher than 10, and (c) the presence of extraintestinal manifestations. PMID- 2561107 TI - [Changes in the adrenoreceptor function in mountaineers with right ventricular hypertrophy]. PMID- 2561108 TI - Protein:DNA interactions at chromosomal loop attachment sites. AB - We have recently identified an evolutionarily conserved class of sequences that organize chromosomal loops in the interphase nucleus, which we have termed "matrix association regions" (MARs). MARs are about 200 bp long, AT-rich, contain topoisomerase II consensus sequences and other AT-rich sequence motifs, often reside near cis-acting regulatory sequences, and their binding sites are abundant (greater than 10,000 per mammalian nucleus). Here we demonstrate that the interactions between the mouse kappa immunoglobulin gene MAR and topoisomerase II or the "nuclear matrix" occur between multiple and sometimes overlapping binding sites. Interestingly, the sites most susceptible to topoisomerase II cleavage are localized near the breakpoints of a previously described illegitimate recombination event. The presence of multiple binding sites within single MARs may allow DNA and RNA polymerase passage without disrupting primary loop organization. PMID- 2561109 TI - Y chromosomal fertility genes of Drosophila: a new type of eukaryotic genes. AB - The Y chromosomal fertility genes of Drosophila are required for sperm differentiation. They are active only in primary spermatocytes where they form giant lampbrush loops. The molecular structure of these genes was investigated and revealed an unusual composition of DNA. Short, tandemly repeated sequence clusters are interrupted by longer and more heterogeneous sequences, which probably all represent transposable elements. No indication of the presence of protein-coding regions has been found within the fertility genes. However, the lampbrush loops bind site-specific proteins recognized by immunofluorescence techniques. This, together with other experimental data, led to the hypothesis that the Y chromosomal genes have a function in binding chromosomal proteins. The data and arguments in support of this gene model are summarized in this paper. PMID- 2561110 TI - Repair of base-base mismatches in simian and human cells. AB - Mismatched heteroduplexes arise as intermediates of several dissimilar genetic processes. The outcome of these genetic events will therefore be influenced by the efficiency and specificity of mismatch repair. We have studied the correction of base-base mispairs in simian and human fibroblasts by transfecting the cells with derivatives of SV40 DNA, each harboring a single mispair in a defined orientation. Analysis of plaques revealed that correction efficiencies for homomispairs followed the pattern G.G greater than C.C greater than or equal to A.A greater than T.T. Repair bias was influenced by flanking sequences. Correction efficiences for heteromispairs followed the pattern of G.T greater than A.C greater than C.T greater than A.G and repair favored the retention of G + C by a substantial margin. This repair specificity could lead to a gene conversion bias favoring the accumulation of G + C in sequences subject to high levels of recombination or unequal exchange. PMID- 2561111 TI - Amino acid similarities to other proteins offer insights into roles of UmuD and UmuC in mutagenesis. AB - The products of the umuD and umuC genes are required for most uv and chemical mutagenesis in Escherichia coli. The genes are organized in an operon that is repressed by LexA and regulated as part of the SOS response. The umuD protein shares homology with the carboxyl-terminal domain of LexA. Genetic evidence now indicates that RecA-mediated cleavage activates UmuD for its role in mutagenesis. The COOH-terminal fragment of UmuD is both necessary and sufficient for this role. Similarities of UmuD to gene 45 protein of bacteriophage T4 and of UmuC to gene 44 protein and gene 62 protein suggest possible roles for UmuD and UmuC in mutagenesis that are supported by preliminary evidence. PMID- 2561112 TI - The biology and exploitation of the retrotransposon Ty in Saccharomyces cerevisiae. AB - Retrotransposons are a widely distributed group of eukaryotic mobile genetic elements that transpose through an RNA intermediate. The element is transcribed into RNA, and this RNA is reverse transcribed into a DNA copy capable of insertion into many different chromosomal locations. Maturation of proteins and reverse transcription take place within noninfectious intracellular viruslike particles. We have studied the element Ty, which is found dispersed in the genome of the yeast Saccharomyces cerevisiae. The frequency of Ty element transposition is normally quite low but can be greatly increased by expressing an element from a strong promoter. We have used the ability to control the level of Ty transposition to investigate the functions of Ty proteins, the regulation of Ty transposition, and the exploitation of Ty elements as insertional mutagens in yeast. The information gained from these experiments should be applicable to the study of retrotransposons found in multicellular organisms. PMID- 2561113 TI - Mobile genetic elements in Drosophila melanogaster (recent experiments). AB - Recent data obtained in the authors' laboratories concerning the behaviour of mobile genetic elements of Drosophila melanogaster are reviewed. It was found that the mobile element jockey represents the typical LINE element. It is efficiently transcribed in D. melanogaster cells in flies and in culture. Transcription is initiated from the +1 nucleotide of jockey and depends on an internal promoter. This is the first case of an internal promoter being used by RNA polymerase II. Several events which take place during the transposition bursts in ctMR2 family of strains were described. Among them are the removal of mobile dispersed genetics (mdg) elements (with solo long terminal repeat (LTR) remaining at the site of excision), complete removal of an mdg element, and reinsertion of the same mdg to the same place either in the presence or in absence of solo LTR sequence. Finally, the formation of deletions was observed. A 462-bp deletion destroying the white locus can be further repaired (w+ reversion). Thus, transposition bursts include many different genetic events. A novel system of prolonged genome destabilization was described. It depends on mobilization of a new mobile element called Stalker. After certain crosses Stalker actively moves for dozens of generations giving rise to large numbers of insertion mutations. Several novel genes were detected using mobilized Stalker. They include a modifier of mdg4 and six enhancers of yellow mutations. PMID- 2561114 TI - Further studies on the influence of dibutyryl cAMP, theophylline and prostaglandin E1 on composition and biosynthesis of phospholipids in Microsporum gypseum. AB - Exogenous supplementation of dibutyryl cAMP and cAMP modulators like theophylline and prostaglandin E1 in the growth medium of Microsporum gypseum lead to increase in the levels of phosphatidylcholine and lysophosphatidylcholine and thereby in total phospholipid content. These observations were further confirmed by the increased incorporation of [32P]orthophosphoric acid into total phospholipid and [14C]choline into phosphatidylcholine and lysophosphatidylcholine. The activity of sn-glycerol-3-phosphate acyltransferase, the enzyme involved in phospholipid synthesis, was stimulated in the presence of dibutyryl cAMP, theophylline and PGE1 supporting the increased synthesis of phospholipids. PMID- 2561115 TI - Basic fibroblast growth factor does not initiate mitogenic signalling via phosphoinositide hydrolysis in PC12 cells. AB - Basic fibroblast growth factor (b FGF) was found to be equally potent mitogen as compared to alpha-thrombin to reinitiate DNA synthesis in quiescent PC12 cells. Whereas thrombin was found to be an activator of phospholipase C as judged by a rapid increase in the formation of inositol triphosphate, inositol biphosphate and a massive accumulation of inositol phosphate when 20 mM LiCl was present as an inhibitor of inositol mono phosphatases, basic FGF failed to induce the breakdown of the polyphosphoinositides in quiescent PC12 cells to any appreciable levels, however, a simultaneous increase in the level of diacylglycerol was observed. b FGF also failed to stimulate protein kinase C which is believed to be activated by diacylglycerol. It is therefore concluded that bFGF receptor mediated 'signalling is not via phospholipase C activation and bFGF's early mitogenic responses and DNA synthesis are initiated independent of the inositol lipids and protein kinase C activation. Thus bFGF must have its own unique signal transducing mechanism independent of inositol pathways. PMID- 2561116 TI - Possible interaction of potassium embelate, a putative analgesic agent, with opiate receptors. AB - The in vivo studies have been carried out in the rat brain for characterization of binding sites for potassium embelate (ex: Embelia ribes) a potent centrally acting analgesic compound. The results indicate that mixed mu and kappa binding sites in the brain may be involved in the analgesic action of this compound. PMID- 2561117 TI - Effects of aspirin and paracetamol on ATPases of human fetal brain: an in vitro study. AB - In vitro effects of aspirin and paracetamol at the doses 200, 400, 600, 800 nmole/mg protein on ATPases activity were studied in the cerebrum and cerebellum of human fetus covering the age range from 10 weeks to 32 weeks of gestation. Both aspirin and paracetamol inhibit Na+K+ ATPase and Mg2+ ATPase in a dose dependent manner. The inhibition of Na+K+ ATPase and Mg2+ ATPase activity which may affect the release and uptake of biogenic amines in CNS, hinders the maturation of human fetal brain. PMID- 2561118 TI - Serum leucine aminopeptidase estimation: a sensitive prognostic indicator of invasiveness in breast carcinoma. AB - Serum Leucine Aminopeptidase (LAP) was studied in 25 patients of breast tumors, and compared with 50 age matched controls. Clinical staging was done in 13 cases of breast carcinoma. Serum LAP was increased but not significantly in stage I (P less than 0.1), while in stage II & III, significant increase was observed (P less than 0.05), but a sharp peak was noted in stage IV with metastasis in liver (P less than 0.001). Serum LAP values were correlated with histological sub-types of breast carcinoma. Highest serum LAP values were observed in filtrating duct carcinoma with productive fibrosis (P less than 0.01), followed by medullary carcinoma and the enzyme values were slightly decreased in intraductal carcinomas. PMID- 2561119 TI - Clinical experience of Oka-strain live varicella vaccine. PMID- 2561120 TI - The effect of thyroid hormones on the membrane adenylate cyclase activity in sickle red blood cells in vitro. AB - A comparative study of thyroid hormone responsive membrane adenylate cyclase activity has been carried out on normal (Hb-AA) and sickle cell (Hb-SS) subjects. Thyroid hormones (T3 and T4) and selected analogues (TRIAC and DLT) enhanced adenylate cyclase activity in Hb-AA membranes. The interaction of Hb-SS membranes with the hormones and analogues was not significant except for T3 which moderately stimulated Hb-SS membrane adenylate cyclase. We suggest that circulating irreversibly sickled cells, in view of their membrane phospholipid defect, probably contribute to the low response of membrane adenylate cyclase to effector stimulation. PMID- 2561122 TI - Hemopoietins and leukemia. AB - This article summarizes research relevant to the possible association between aberrant production of hemopoietins or hemopoietin-activated signals and the neoplastic growth of leukemic cells. PMID- 2561121 TI - Erythropoietin: biology and clinical use. AB - Red cell production in vertebrates is controlled by a glycoprotein hormone known as erythropoietin (Ep), which is produced by the kidney in response to hypoxia and acts on the marrow to selectively stimulate erythropoiesis. The gene for Ep has recently been cloned, and highly pure recombinant human Ep (rHuEp) is now available in considerable quantity. This has led to a better understanding of many aspects of Ep biology and to clinical trials in humans. The amino acid sequence of Ep is now completely known, and the protein portion of the natural hormone and the recombinant product are identical. Both the natural hormone and rHuEp produced in Chinese hamster ovary cells are heavily glycosylated in a very similar manner. This glycosylation is not necessary for in vitro activity but is required for activity in vivo. Radioimmunoassays (RIAs), which use labeled rHuEp, have been developed and are sufficiently sensitive to measure normal plasma levels. However, since Ep exists in plasma in several forms that vary in their immunologic and biologic activities, the ability of a RIA to provide information on the pathogenesis of clinical disease may be limited and should be referenced to the polycythemic mouse assay. The kidney's role in the production of Ep has been greatly clarified. Studies using probes to Ep mRNA have shown that Ep is primarily made in the kidney and secreted as the intact hormone. Moreover, renal secretion appears to be regulated by the rate of synthesis of the hormone, which in turn is dependent on the rate of synthesis of Ep mRNA. The cells that produce Ep have been identified as peritubular interstitial cells that may be endothelial in origin. The initiating mechanism for hormone production appears primarily to involve recruitment of additional cells rather than increased production by individual cells. Ep primarily acts on the marrow to stimulate the growth and maturation of early cells in the erythroid lineage that are known as the burst forming unit-erythroid (BFU-E) and colony-forming unit-erythroid (CFU-E). The BFU E is a very early cell closely related to the pluripotent stem cell, while the CFU-E is a later cell close to the first recognizable erythroblast.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2561123 TI - Pseudomyxoma peritonei. A report of three cases. AB - Pseudomyxoma peritonei is an infrequently encountered clinical entity which presents mainly with abdominal distension. We present here our experience with three such cases and emphasize on the aggressive combined operative and chemotherapeutic approach in this disease. PMID- 2561124 TI - Clinical profile and electrodiagnostic study of peripheral neuropathy. AB - A clinical analysis and electrophysiological study was performed in 40 cases of peripheral neuropathy. Motor nerve conduction velocity (MNCV) and electromyography (EMG) were also recorded in 20 healthy volunteers. Twelve cases were of diabetic neuropathy (DN), nine of Guillain-Barre syndrome (GBS), eight of idiopathic, six of leprous, three of toxic neuropathy and one each of acute intermittent porphyria (AIP) and carcinomatous neuropathy. The onset was insidious in a majority of the cases (70%), the presenting symptom being paraesthesia (27.5%). A variable sensory loss was the commonest finding (77.5%). Two patients each had evidence of cranial nerve involvement and affection of urinary bladder and bowel. In both DN and GBS there was a significant reduction in the MNCV especially in the peroneal nerves. In DN 41.6% had evidence of decreased motor unit potentials and 16.6% showed spontaneous fibrillatory activity. For GBS these figures were 77.7% and 8.3% respectively. The peripheral nerve involvement was variable in leprosy and the EMG was normal in all, but one case. PMID- 2561125 TI - Coxsackie virus and rheumatic fever. A correlative study. AB - Twenty-one patients of whom 13 had acute rheumatic fever and 8 had recurrence of rheumatic fever were studied for the evidence of coxsackie B viral infection using neutralisation test. A significant rise was noted in 17 cases (81%) and two cases had very high initial titre of neutralising antibodies to coxsackie B viruses type 1 to 6. Mixed infection with more than one serotype was seen in 11 cases. Coxsackie B2 was the commonest type and 14 patients had antistreptolysin 'O' anti-bodies. The high incidence of coxsackie B viral infection in rheumatic fever and the coexistent streptococcal infection and their relationship are discussed. PMID- 2561126 TI - New insights into the evolutionary growth of tumors revealed by Southern gel analysis of tumors genetically tagged with plasmid or proviral DNA insertions. PMID- 2561127 TI - Regulatory signals affecting a selective loss of mRNA in Dictyostelium discoideum. AB - We identified signals that affect mRNA levels complementary to a gene that is highly expressed in vegetative Dictyostelium discoideum cells. This gene has been cloned as cDNA in the plasmid pcD-D2. The level of transcripts homologous to pcD D2 fell dramatically in strain XP55 during the aggregation stage of development when cells differentiate on agar. The level, however, did not fall simply as a result of starvation or aggregation-specific cell contact. Rather, before the level is reduced cells must be deprived of amino acids and cyclic AMP administered in amounts and at intervals in pulses to mimic cyclic AMP signal relay in aggregation. This effect can be blocked either with cyclic AMP-S (a non hydrolysable cyclic AMP analogue) or adenosine, both of which prevent cyclic AMP binding to the cyclic AMP cell surface receptor. It is also blocked in 'frigid' aggregation-deficient mutants HC85 and HC112 known to be defective in a G alpha protein. We conclude that the transcript level is balanced by positive nutritional signals acting against negative signals transduced in part through a cell surface cyclic AMP receptor. PMID- 2561128 TI - Bradykinin receptor number and sensitivity to ligand stimulation of mitogenesis is increased by expression of a mutant ras oncogene. AB - Bradykinin is a nine amino acid peptide of the kinin family believed to play a role in pain mediation and in the regulation of blood pressure, fluid balance and smooth muscle contraction. Here we demonstrate that bradykinin is also a potent mitogen for a mutant Ha-ras-transfected cell line, Rat 13. The Rat 13 cells display two binding sites for bradykinin: a moderate number (52,000) of high affinity sites (Kd = 4.9 nM) co-exist with a much smaller number (1100) of very high-affinity sites (Kd = 2.7 pM). Ligand binding stimulates mitogenesis through the lower affinity receptors, which are classified as B2. These receptors are down-regulated in response to ligand. In contrast, Rat 1 cells (the cell line from which Rat 13 was derived) have only 4000 receptors per cell in total and respond weakly to bradykinin as a mitogen. Thus, expression of a mutant ras protein in Rat 13 cells increases their expression of the bradykinin receptor and their sensitivity to ligand stimulation of mitogenesis. Additional binding studies demonstrate that human A431 epithelial cells and Swiss mouse 3T3 fibroblasts also possess high-affinity sites for bradykinin. PMID- 2561129 TI - Possible function of matrix proteins in fluoride incorporation into enamel mineral during porcine amelogenesis. AB - The present study was undertaken to elucidate the mechanism of fluoride incorporation into secretory enamel mineral, with porcine enamel used as a model. Although the fluoride content in the enamel varied greatly among the animals, we observed that the fluoride-to-calcium ratio in the enamel tissue was maximal at the beginning of the secretory stage; the F/Ca ratio decreased (and leveled off) with the advancement of mineralization. In vitro work showed that some of the fluoride in the secretory enamel tissue was removed with the extraction of organic matter, mostly amelogenins. Furthermore, coating hydroxyapatite crystals with enamel matrix proteins resulted in a retardation of fluoride incorporation into the crystals when exposed to fluoride solutions, as a result of an inhibition of apatite reprecipitation. We also confirmed that the growth kinetics of fluoridated apatite onto HA seeds decreased with increasing coverage of the seed surface with the enamel proteins. All the results of the present study strongly suggest that the fluoride incorporation into enamel mineral during the secretory stage may be regulated by the kinetics of mineralization, which is highly dependent on the driving force for precipitation and the presence of proteinaceous inhibitors, mainly amelogenins. PMID- 2561130 TI - Suggested mechanism by which a film of agar gel purifies and concentrates rotavirus from a faecal suspension before electron microscopy. PMID- 2561131 TI - An evolutionary relationship between the ColE5-099 and the ColE9-J plasmids revealed by nucleotide sequencing. AB - The nucleotide sequence of a 1124 bp fragment of the ColE5-099 plasmid which encodes colicin E5 immunity, a lys gene involved in colicin release from the host cell, and the 3' end of the colicin E5 structural gene has been determined. Open reading frames corresponding to the three genes have been located by analogy with similar sequences from other E colicin plasmids. The location of these open reading frames corresponds with the position of the genes as determined by subcloning and transposon mutagenesis. The amino acid sequence of the carboxy terminal 107 amino acid residues of the colicin E5 gene shows no homology with any other E colicin, suggesting a different mode of action in killing sensitive cells. A comparison of the nucleotide sequence of this region of the ColE5-099 plasmid with that of the equivalent region of the ColE9-J plasmid suggests a close evolutionary relationship between these two plasmids. PMID- 2561132 TI - Nutritive utilization of calcium in rats: effects of dietary fat components and vitamin D3 on intestinal resected rats. AB - The nutritive utilization of calcium was studied in adult rats in which 50% of the distal small intestine (DSI) had been resected and in sham-operated controls one month and three months after the operation. Resection of half the DSI reduced the digestive utilization of Ca as reflected by mineral content in bone. Three months after resection, nutritive utilization of Ca had still not recovered. Feeding the resected rats with a diet in which fat content consisted of equal parts of medium-chain triglycerides (MCT), sunflower seed oil, and olive oil failed to improve nutritive utilization of Ca after one or three months in comparison with a diet containing olive oil as the only source of lipids. Supplementation with vitamin D3 (0.04 mg/100 g diet) enhanced nutritive utilization of Ca in resected rats after one month, the beneficial effect becoming much more patent after three months. At the dose used, vitamin D3 favored calcium deposition in bone tissue. Serum levels of Ca remained unchanged under all experimental conditions, both one month and three months after 50% DSI resection. PMID- 2561133 TI - Protein-binding properties of 22-oxa-1 alpha,25-dihydroxyvitamin D3, a synthetic analogue of 1 alpha,25-dihydroxyvitamin D3. AB - Protein binding properties of 22-oxa-1 alpha,25-dihydroxyvitamin D3 (22-oxa-1,25 D3), a synthetic analogue of 1 alpha,25-dihydroxyvitamin D3 (1,25-D3), were compared with those of vitamin D3 derivatives. The order of binding affinity to the chick embryonic intestinal receptor was 1,25-D3 greater than 22-oxa-1,25-D3 greater than 25-hydroxyvitamin D3 (25-D3) greater than 24R, 25-dihydroxyvitamin D3 (24, 25-D3) greater than vitamin D3 (D3), while that to the rat plasma vitamin D-binding protein (DBP) was 25-D3 greater than 24,25-D3 greater than D3 greater than 1,25-D3 greater than 22-oxa-1,25-D3. The binding potencies of 22-oxa-1,25-D3 to the receptor and DBP were about 1/8 and 1/600 of the respective values of 1,25 D3. When the distribution of the tritiated compounds in human plasma components was examined by an in vitro polyacrylamide gel electrophoretic method, [3H]-22 oxa-1,25-D3 was found to bind only to the lipoproteins including chyromicron. These results suggest that the replacement of a carbon atom into an oxygen atom in the side chain structure of 1,25-D3 results significant decrease in the binding affinity to DBP and that 22-oxa-1,25-D3 is transported as a complex-form not with DBP but with lipoprotein to the target tissues. PMID- 2561134 TI - Potentiated endothelin-1-induced phosphoinositide hydrolysis in atria and mesenteric artery of DOCA-salt hypertensive rats. AB - The effect of endothelin-1 on the phosphoinositide pathway was studied in slices of atria and mesenteric artery from normotensive and hypertensive (DOCA-salt) rats. Endothelin-1 induced a dose-dependent increase in inositol monophosphate production in both groups, but the reactivity of the phosphoinositide pathway was enhanced in DOCA-salt hypertensive rats. Endothelin-1 was more potent than noradrenaline and activation by these two agonists combined was additive. The phosphoinositide activation induced by endothelin-1 was independent of extracellular calcium, and was not prevented by nifedipine treatment or by removing calcium from the incubation medium. Endothelin-1 is a potent direct activator of the phosphoinositide pathway in cardiovascular tissues, and this effect is potentiated in DOCA-salt hypertension. PMID- 2561135 TI - Stimulation of Na(+)-H+ exchange, the Na(+)-K+ pump and Na+,K+,Cl- cotransport by endothelin-1 in cultured vascular smooth muscle cells. AB - Endothelin-1 concentrations higher than 10(-10) mol/l were able to strongly stimulate Na(+)-H+ exchange, the Na(+)-K+ pump and the Na+,K+Cl- cotransport system in A10 vascular smooth muscle cells. The stimulation of Na(+)-H+ exchange was more marked in fresh than in H(+)-loaded cells, suggesting an increase in the apparent affinity for internal H+. Pump stimulation appeared to be secondary to sodium entry through Na(+)-H+ exchange because it was absent in (1) Na(+)-loaded cells and (2) in the presence of ethyl-isopropylamiloride (EIPA). Stimulation of cotransport was blocked by indomethacin, suggesting the involvement of a cyclo oxygenase product. In conclusion, the action of endothelin-1 in vascular smooth muscle induces the onset of negative feedback mechanisms which enhance the ability of the vascular cells to regulate disturbances in Na+,K+ and Cl- contents. PMID- 2561136 TI - Cortisol-stimulated phosphoinositide metabolism in vascular smooth muscle cells: a role for glucocorticoids in blood pressure control? AB - Physiological concentrations of cortisol (0.02-5.0 micrograms/ml) stimulate the phosphoinositide breakdown in cultured smooth muscle cells from rat aorta. After a 15-min stimulation of the cells with various concentrations of cortisol, the inositol-1,4,5-triphosphate concentrations showed a dose-dependent increase. This phenomenon was qualitatively similar but quantitatively more sensitive in smooth muscle cells from spontaneously hypertensive rats (SHR) than in vascular smooth muscle cells from normotensive animals. When the antihormone RU 486 was added before the vascular smooth muscle cells were stimulated with cortisol, the inositol-1,4,5-triphosphate production was reduced to 50%. PMID- 2561137 TI - Platelet cyclic AMP in essential hypertensive and normotensive offspring. AB - Essential hypertension is accompanied by several modifications to platelet metabolism suggesting hyper-reactivity to various aggregating agents. As the platelet response is mediated by both cytosolic free calcium, which is stimulatory, and cyclic (c)AMP, which is inhibitory, this hyper-reactivity may be caused by a modification in cAMP metabolism. We therefore determined cAMP in unstimulated platelets from 19 patients with essential hypertension and 27 age matched normotensive subjects, nine with and 19 without a family history of hypertension. The platelet cAMP content was reduced in the essential hypertensives and in the normotensives with a positive family history by 37.5% and 42%, respectively (P less than 0.001 for both). Platelet cAMP was inversely correlated with diastolic blood pressure (P = 0.036). After prostaglandin (PG) E1 stimulation, the platelet cAMP content remained lower in the patients with essential hypertension than in the normotensive subjects, whatever their hypertensive heredity. The rises in cAMP caused by inhibition of phosphodiesterase by 7-bromo-1,5-dihydro-3,6-dimethylimidazo-[2,1-b]quinazolin-2[ 3H]-one (Ro 15-2041) were similar in the three groups. These results indicate that cAMP, the platelet inhibitory messenger, is reduced in hypertensive patients and in their normotensive offspring and may affect the various platelet abnormalities previously described in this disease. PMID- 2561138 TI - Renal sodium handling abnormalities in hypertensive and normotensive patients with a family history of hypertension. AB - We measured the effective renal plasma flow, the glomerular filtration rate and the fractional excretion of sodium under low and high sodium intakes in the normotensive offspring of hypertensives and in essential hypertensive patients. During the high sodium intake, 57% of the offspring and 42% of the hypertensive patients showed an increase in effective renal plasma flow and fractional excretion of sodium. In the remaining hypertensives, enalapril improved the renal haemodynamic and sodium-handling responses. Enalapril also reduced the blood pressure in all hypertensives, whether they showed a normal or an abnormal response to a high sodium intake. In the hypertensive patients with an abnormal response to a high sodium intake, the intra-erythrocyte sodium content was higher than in the normal responders. Thus, abnormal renal haemodynamic and sodium handling responses to a high sodium intake were observed in 50% of the hypertensive patients and the offspring. The high intra-erythrocyte sodium content observed in the hypertensive patients suggests that extrarenal alterations in sodium handling were present. PMID- 2561139 TI - Brain corticosteroid receptors and regulation of arterial blood pressure. AB - A single intracerebroventricular injection of 10 ng of the mineralocorticoid aldosterone in normotensive male Wistar rats induced a transient increase in blood pressure. Corticosterone produced this response only at a 100-fold higher dose. In contrast, a decrease in systolic blood pressure occurred following the intracerebroventricular administration of a selective glucocorticoid agonist (Ru 28362; 10 ng). The intracerebroventricular administration in normotensive and adrenalectomized, corticosterone-replaced, male rats of an antimineralocorticoid (Ru 28318; 10 ng) decreased systolic blood pressure, while an antiglucocorticoid (Ru 38486; 10 ng) caused an increase. These data suggest that adrenal steroids affect central mechanisms underlying cardiovascular regulation by binding to central mineralo- and glucocorticoid receptors. PMID- 2561140 TI - Circulating prorenin and renin in response to intravenous adrenocorticotrophic hormone in essential hypertension. AB - Plasma prorenin and renin changes after a bolus injection of 25 U intravenous adrenocorticotrophic hormone (ACTH, synacthen) were studied in seven untreated uncomplicated essential hypertensives over a period of 24 h. Plasma prorenin did not change significantly during the study, whereas renin after 24 h was higher than at baseline (4.3 +/- 0.6 versus 2.3 +/- 0.9 ng angiotensin I (Ang I)/ml per h, P less than 0.01). We conclude that endogenous glucocorticoid stimulation induced by exogenous ACTH and ACTH itself seem to induce a secondary or tertiary rather than a primary effect on the human renin gene. PMID- 2561141 TI - Prediction of response to antihypertensive therapy with enalapril and nifedipine. AB - We evaluated the usefulness of an integrated concentration-effect modelling technique in predicting the long-term response to antihypertensive therapy with enalapril and nifedipine. Two groups of essential hypertensives were given monotherapy with 20 mg nifedipine twice a day (n = 14) or 20 mg enalapril once a day (n = 13), and were studied following the administration of the drugs and after at least 6 weeks' treatment. For both drugs the predicted responses (predose and 4 h postdose) were in close agreement with the observed responses. With enalapril the observed and predicted profiles over a 12-h study period were well correlated in all subjects. In contrast, with nifedipine, although there was generally good agreement, the model over-predicted the profile of response in one patient and under-predicted the profile in one patient. Thus, there is evidence that application of concentration-effect analysis is useful in predicting the steady-state antihypertensive effect from the first dose-response to the drug. PMID- 2561142 TI - Central and peripheral opiate receptors appear to be activated during controlled haemorrhagic hypotension. AB - The present study investigated the question of whether peripheral or central opiate receptors are activated during controlled haemorrhagic hypotension. In anaesthetized Wistar rats, blood pressure was reduced by steps, by bleeding, to 80, 60 and 40 mmHg. Subcutaneous administration of 1 mg/kg of naloxone and of methyl naloxone-Br, an analogue of naloxone, which does not readily cross the blood-brain barrier, significantly elevated the bleeding volume at the 40-mmHg blood pressure level. A dose of 10 mg/kg of methyl naloxone-Br had no effect on the bleeding volume. We therefore conclude that during haemorrhage, endogenous opioid peptides activate both central and peripheral opiate receptors, thereby exerting a hypotensive influence. PMID- 2561143 TI - The haemodynamic effects of two angiotensin converting enzyme inhibitors, enalaprilat and zofenoprilat, in the rat: evidence for the involvement of bradykinin. AB - Since angiotensin converting enzyme (ACE) metabolizes bradykinin, the hypotensive effect of ACE inhibitors could be partly due to an increased bradykinin activity. We therefore investigated the influence of HOE K86-4321 [D-Arg-(Hyp2-Thi5,8 DPhe7)-bradykinin], a selective bradykinin-2 receptor antagonist, on the effects of enalaprilat (0.3 and 3.0 mg/kg) and zofenoprilat (0.1 and 1.0 mg/kg) on the heart rate, mean arterial blood pressure, cardiac output and total peripheral resistance in rats. Both enalaprilat and zofenoprilat reduced mean arterial pressure (from 110 +/- 7 to 85 +/- 6 and from 108 +/- 9 to 72 +/- 9 mmHg, respectively; P less than 0.05) and total peripheral resistance (from 515 +/- 35 to 413 +/- 29 and from 495 +/- 45 to 310 +/- 25 x 10(-3) mmHg/litre per min per kg, respectively; P less than 0.05); the heart rate and cardiac output changed little. In the presence of HOE K86-4321, which by itself did not affect the haemodynamic variables measured, the effects of the two ACE inhibitors were significantly reduced. These results suggest that bradykinin-2 receptor-mediated vasodilation, although not involved in blood pressure regulation, influences the reduction in blood pressure induced by enalaprilat and zofenoprilat in normotensive rats. Furthermore, at comparable ACE-inhibiting doses, zofenoprilat was more effective in reducing mean arterial pressure, which might be related to the presence of a sulphydryl group. PMID- 2561144 TI - Blunting of atrial natriuretic factor response to volume expansion by benazepril in hypertensive patients. AB - To verify the hypothesis that the angiotensin converting enzyme (ACE) level may affect the metabolism of circulating atrial natriuretic factor (ANF), the acute and chronic effects of benazepril on plasma ANF levels were studied in hypertensive patients under basal conditions and in response to acute volume expansion. Ten essential hypertensives entered a double-blind crossover study, and were randomly allocated either to placebo or to 10 mg benazepril orally once a day for 2 days; after a placebo washout period of 2 days the groups were crossed over. On the second day of each crossover period, volume expansion was induced by infusing 1 litre saline in 30 min, and blood samples for ANF measurements were drawn at times -5, 0, 5, 15, 30, 35, 40, 50 and 60 min. Oral benazepril at 10 mg/day was then given to all patients for 4 weeks, and the volume expansion with saline was repeated. After the 2-day acute benazepril treatment, blood pressure fell from 166.1 +/- 3.6/105.1 +/- 0.9 to 140.1 +/- 4.6/85.6 +/- 2.1 mmHg (P less than 0.01 for both systolic and diastolic blood pressure), whereas ANF fell from 29.4 +/- 3.6 to 24.1 +/- 3.7 pg/ml (NS) after the acute benazepril treatment and to 17.7 +/- 3.6 pg/ml (P less than 0.01) after the chronic benazepril treatment. The volume expansion itself did not induce significant changes in mean arterial pressure, either during the placebo treatment or during the acute chronic benazepril treatment. The rise in ANF values in response to saline infusion during placebo was prompt, beginning at min 15 and reaching a maximum at min 40.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561145 TI - Comparison of the acute and chronic antihypertensive effect of two once-daily doses of spirapril by invasive twenty-four-hour ambulatory blood pressure monitoring. AB - Spirapril is a new non-sulphydryl angiotensin converting enzyme (ACE) inhibitor which is eliminated mainly through the liver. In a placebo-controlled study the acute and chronic (2 weeks) antihypertensive effects of two doses of spirapril (12.5 and 25 mg) were assessed by invasive 24-h ambulatory blood pressure monitoring in two groups of eight hypertensive subjects. After the first doses of 12.5 and 25 mg, blood pressure rapidly decreased, with a nadir after 4-6 h. At this time, mean arterial pressure was decreased by 24 +/- 2% with the 12.5 and 19 +/- 1% with the 25-mg dose. The antihypertensive response was sustained for almost the whole 24-h period with the 25-mg dose, but it was attenuated during the second half of the 24-h blood pressure monitoring with the 12.5-mg dose, leaving a residual antihypertensive effect of 6 +/- 2% 24 h after dosing. Compared with the response to the first dose, the antihypertensive response after 2 weeks of treatment was similar to that following the 12.5-mg dose, but with the 25-mg dose the response during the second half of the 24-h monitoring period, especially during the night, was attenuated. The reason for this attenuated antihypertensive response during prolonged ACE inhibition remains to be established. PMID- 2561146 TI - The incidence of cough during treatment with angiotensin converting enzyme inhibitors. AB - The incidence of cough during treatment with angiotensin converting enzyme (ACE) inhibitors was studied using retrospective analysis of outpatient records and a questionnaire; for a more precise evaluation, all reported cases of cough were reviewed according to criteria for the operational assessment of side effects, and those found unrelated were excluded. Cough during treatment with ACE inhibitors appears to be more frequent than previously recorded and substantial differences between patients treated with captopril or enalapril seem unlikely. PMID- 2561147 TI - Angiotensin converting enzyme inhibition in normotensive type II diabetics with persistent mild proteinuria. AB - To evaluate the effect of enalapril on proteinuria, 16 normotensive type II diabetics with persistent proteinuria were studied. At random, the patients were allocated to enalapril (5 mg once a day) or placebo, in a double-blind fashion, for 12 months. Blood pressure, heart rate, urinary albumin excretion, plasma renin activity and aldosterone, blood glucose, serum fructosamine, urine urea and body weight were checked monthly during the run-in period and every 2 months during the treatment period. The kidney function was studied at the beginning of the study and during the sixth and 12th months. Enalapril decreased urinary albumin excretion in our patients in the absence of any effect on blood pressure and kidney function. Our data may be interpreted on the basis of a direct vascular effect of enalapril that is probably related to a tissue mechanism consisting of reduced angiotensin formation, increased kinins, or both, or of other unknown factors. PMID- 2561148 TI - A mechanical stress increases sodium ion content in vascular smooth muscle cells through a calcium-dependent pathway: prevention by cicletanine via a cyclo oxygenase product. AB - The non-laminar (rather turbulent) flow induced by cell washings was able to reversibly increase the sodium ion (Na+) content in cultured A10 aortic smooth muscle cells. Similar changes, although to a lesser extent, were observed in cardiocytes but not in fibroblasts, erythrocytes, thymocytes or macrophages, suggesting that the changes are specific to excitable cells. The increase in vascular sodium content had the following properties: (1) It was inhibited by nitrendipine; (2) it was accompanied by an increase in the free cytosolic Ca2+ content; (3) it was unable to stimulate the sodium pump; and (4) it reflected the qualitative and quantitative composition of the incubation media. These observations suggested that a non-laminar flow is able to open potential dependent calcium channels, with secondary internalization of high amounts of extracellular ions. These ionic perturbations were blocked by low concentrations of cicletanine; the half-maximal inhibitory concentration (IC50) was about 10(-9) mol/l on internal sodium. The protective effects of cicletanine were inhibited by indomethacin, suggesting that they are mediated by a cyclooxygenase metabolite, perhaps prostacyclin. Captopril and diuretic drugs such as hydrochlorothiazide, furosemide, spironolactone or acetazolamide were unable to protect vascular cells against the harmful effects of cell washings. PMID- 2561149 TI - Nadolol prevents the exercise-induced rise in lymphocyte beta-receptor number in borderline hypertension. AB - Thirteen borderline hypertensives were investigated at rest and during dynamic exercise, before and after therapy with nadolol (40-80 mg/day for 7-28 days), in order to evaluate regulation of the number of lymphocyte beta-receptors. Systolic blood pressure and the heart rate were measured before and after 15 min of bicycle exercise, both with and without nadolol therapy; blood samples were withdrawn for adrenaline, noradrenaline and lymphocyte beta-receptor determinations. Nadolol induced a significant decrease in systolic blood pressure and the heart rate at rest, while plasma catecholamines and lymphocyte beta receptors did not change significantly. Of the physiological responses to dynamic exercise (increases in systolic blood pressure, heart rate, plasma noradrenaline levels and adrenaline and lymphocyte beta-receptors), only the rise in beta receptors was entirely prevented, and the increase in the heart rate was significantly attenuated by nadolol. It is suggested that the lack of a rise in the number of beta-receptors during exercise may contribute to the blunted exercise-induced tachycardia in patients taking nadolol. PMID- 2561150 TI - Alpha 2- and beta 2-adrenoceptor downregulation in marathon runners. AB - The regulation of platelet alpha 2- and lymphocyte beta 2-adrenoceptor densities by alterations in endogenous catecholamines was examined. In order to activate the sympathetic nervous system eight trained male normotensive subjects carried out a marathon run. Adrenoceptor densities and plasma catecholamine concentrations were measured before and immediately after the run. Platelet alpha 2-adrenoceptor density and lymphocyte beta-adrenoceptor density decreased after the run (P less than 0.05), whereas both plasma noradrenaline and adrenaline concentrations increased (P less than 0.01). Mean arterial pressure decreased (P less than 0.05), and the heart rate increased (P less than 0.001). The data suggest that increases in endogenous catecholamine concentrations cause downregulation of both alpha- and beta-adrenoceptor densities on human blood cells. PMID- 2561151 TI - Properties of alpha 2-adrenoceptors in human myometrium and kidney: similarities with human platelets, but differences in the rat kidney. PMID- 2561152 TI - Increased beta-adrenergic tone enhances arterial compliance in hyperkinetic borderline hypertension. AB - Arterial compliance, assessed by the stroke volume-pulse pressure relationship, was measured in 32 patients with a high cardiac output together with borderline hypertension and 26 control subjects with normokinetic borderline hypertension. Measurements were obtained in the supine position both before and after 0.2 mg/kg intravenous propranolol. Baseline arterial compliance was significantly greater in the hyperkinetic group (P less than 0.01). The inter-group difference was no longer significant after the administration of propranolol, since compliance tended to increase in the normokinetic group and decrease in the hyperkinetic group. Therefore, disparities in vascular beta-adrenergic tone between hyperkinetic and normokinetic borderline hypertensive patients probably contribute to group differences in arterial compliance. PMID- 2561153 TI - Pressor responsiveness, age and hypertension. AB - It has long been accepted that enhanced vascular reactivity is an integral part of the hypertensive state. This study investigates pressor responsiveness to the selective alpha 1-adrenoceptor agonist, phenylephrine, in 62 normotensive and hypertensive subjects aged 20-70 years. Since blood pressure increases with age, it is possible that effects attributed specifically to blood pressure will be confounded by age. The present study showed no correlation between responsiveness and starting blood pressure. However, responsiveness was positively correlated with age. These results suggest that it may be an oversimplification to attribute increased vascular responsiveness to the hypertensive state. PMID- 2561154 TI - Endothelin has potent direct inotropic and chronotropic effects in cultured heart cells. AB - To determine whether endothelin, a highly potent vasoconstrictor peptide, may affect cardiac myocyte contractility directly, we studied the effects of synthetic porcine endothelin-1 in cultured chick embryo ventricular cells. Endothelin-1 had a potent chronotropic effect (EC50 0.17 nmol/l) in spontaneously beating cells. The increase in the beating rate was accompanied by a frequency dependent decrease in the amplitude of contraction. In electrically driven cells (1 Hz), endothelin-1 increased the amplitude of contraction dose-dependently, with an EC50 of 0.3 nmol/l, smaller than that of the calcium channel blocker BAY K 8644 (EC50 3.3 nmol/l), and with an efficacy close to that of BAY K 8644 and isoproterenol. Nicardipine (100 nmol/l) shifted to the right, by two orders of magnitude, the dose-response curve of endothelin-1 for inotropism. These results indicate that endothelin-1 is one of the most potent inotropic agents in cultured cardiac myocytes, and suggest that this effect involves, at least in part, a calcium ion influx through voltage-sensitive calcium channels. PMID- 2561155 TI - Relationship between cyclic nucleotide levels and 5-methyl-6-(4-pyridyl)-2H-1,4 thiazin-3(4H)-one (ZSY-27), a new positive inotropic agent with a vasodilatory action, -induced relaxation of rabbit thoracic aorta. AB - The aim of this investigation was to substantiate the hypothesis that the vasorelaxant effects of 5-methyl-6-(4-pyridyl)-2H-1,4-thiazin-3(4H)-one (ZSY-27) are mediated by accumulation of intracellular cyclic nucleotides as a consequence of inhibition of cyclic nucleotide phosphodiesterase activity. Both activities of adenosine 3',5'-monophosphate-phosphodiesterase (cAMP-PDE) in the presence of ethylene glycol-bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA) and guanosine 3',5'-monophosphate-phosphodiesterase (cGMP-PDE) in the presence of calcium-calmodulin from rabbit thoracic aorta were inhibited in a concentration dependent manner by ZSY-27 (10(-5) - 10(-3) M). The IC50 values for ZSY-27 on cAMP- and cGMP-PDE activity were 2.1 x 10(-4) and 8.8 x 10(-4) M, respectively. Furthermore, ZSY-27 antagonized competitively cAMP-PDE (Ki = 1.9 x 10(-4) M). On the other hand, ZSY-27 exhibited a mixed-type inhibitory pattern, with reduction of both maximum velocity and affinity for the substrate of the cGMP-PDE, with a Ki value of 1.0 x 10(-3) M. Spontaneous myogenic tone of rabbit thoracic aorta was significantly attenuated from 1 min after addition of ZSY-27 (3 x 10(-4) M). Contents of cAMP and cGMP were significantly increased from 1 and 3 min after addition of ZSY-27, respectively. Temporally, relaxant effects of ZSY-27 were associated with increases of cAMP content, but not with that of cGMP content.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561156 TI - A Chinese traditional medicine, juzentaihoto, inhibits the O2- generation by macrophages. AB - Guinea pig peritoneal macrophages generate superoxide anion (O2-), when stimulated with N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP), phorbol myristate acetate (PMA) or ovalbumin complex of guinea pig IgG2 antibody (OA gamma 2). These responses were found to be inhibited by a Chinese traditional medicine, Juzentaihoto (JT). When the inhibition was assayed by the use of fMLP as a stimulant, at least one of the substances responsible for the inhibitory activity of JT was identified to be cinnamic acid. An authentic sample of cinnamic acid also inhibited the O2- generation by fMLP-stimulated macrophages. Cinnamic acid, however, did not inhibit the O2- generation, when macrophages were stimulated with PMA and OA gamma 2. These results indicate that a certain cinnamic acid-inhibitable factor may be involved in the intracellular triggering event(s) initiated by fMLP, leading to activation of the respiratory burst reduced nicotinamide adenine dinucleotide phosphate oxidase, but not in those by PMA and OA gamma 2. PMID- 2561157 TI - Streptomyces nucleotide 3'-pyrophosphokinase are insensitive to stringent control. AB - The synthesis of GTP- and ATP-3'-pyrophosphate by Streptomyces morookaensis nucleotide 3'-pyrophosphokinase was little affected by the presence of a ribosome deacylated tRNA-poly A, G, U mixture in vitro. The Streptomyces pyrophosphokinases thus do not appear, unlike bacterial stringent factors, to play a significant role in the cellular stringent control. PMID- 2561158 TI - A therapeutic profile of metal chelators in the detoxication of methylmercury chloride inhibited acid and alkaline phosphatases in different areas of the central nervous system of rats. AB - Acid and alkaline phosphatase changes in various parts of the central nervous system (olfactory bulbs, cerebral hemispheres, cerebellum, medulla oblongata, and spinal cord) were analyzed during methylmercury chloride (MMC) treatment with a dose of 5 mg/kg/day body weight. The drug was subcutaneously introduced into the animals and the enzymes were analyzed after 2, 7, and 15 days' treatment. One group of animals was treated for seven days and kept without drug for another seven days (withdrawal group). The antagonizing capacities of four chelators, namely, N-acetyl-DL-homocysteine thiolactone (NAHT), D-penicillamine (DPA), glutathione (GSH), and sodium selenite (SEL), were also analyzed in relation to the restoration of enzymes. Study results show a linear inhibition of acid and alkaline phosphatases with increasing duration of MMC treatment. However, the magnitude of enzymatic inhibition is different in different brain areas. After 15 days' treatment, maximum inhibition of acid and alkaline phosphatases was recorded in the spinal cord and cerebellum, respectively. Chelators also exhibited differential recovery of the enzymes in various animal groups, as well as in discrete brain areas. No uniformity in the recovery of the enzymes with chelators was observed. However, study results show that biochemical parameters are good indicators of early recognition of neurotoxicity. PMID- 2561159 TI - Elevation of 3'5' cyclic adenosine monophosphate alters CD3 and CD25 antigens expression in activated T lymphocytes. AB - Modulation of CD3 molecules and expression of receptors for IL-2 (CD25) are pivotal events of lymphocyte activation and proliferation. Knowing the inhibitory effect of cAMP elevating agents on T lymphocyte activation, we investigated the effect of cholera toxin (CT) and dibutyryl cyclic AMP (dbcAMP) on the modulation of the CD3/Ti complex and on the appearance of the CD25 antigen on PHA-activated human lymphocytes. Cytofluorometry analysis of indirectly anti-CD3 labelled cells showed that CT accelerated the disappearance of CD3 molecules and slowed their reappearance. CT or dbcAMP inhibited the expression of CD25 antigen. In both cases, not only the relative number of CD3+ or CD25+ cells decreased, but the number of CD3 or CD25 antigens per cell as well. Exogenous rIL-2 did not reverse the inhibition of IL-2R expression by CT, showing that this effect is independent of the inhibition of IL-2 production already demonstrated. We conclude that augmenting cAMP levels might affect early steps of activation such as antigen receptor modulation, but do affect more profoundly late IL-2 dependent steps especially the autocrine IL-2 pathway of IL-2 receptor upregulation and the production of IL-2. PMID- 2561160 TI - Decrease with senescence in the norepinephrine-induced phosphorylation of myofilament proteins in isolated rat cardiac myocytes. AB - (1) The effects of norepinephrine on protein phosphorylation in isolated rat cardiac ventricular myocytes were determined by autoradiography on 32P-labelled proteins separated by electrophoresis; (2) In cells from young adult rats (6 months old) there was a marked increase due to norepinephrine (10(-8) to 10(-4) M) in the incorporation of 32P into proteins identified on the grounds of molecular weight as troponin I and C-protein: in cells from senescent rats (24 months old) this increase was much attenuated. (3) Age-associated decrements in protein phosphorylation were much diminished when maximally effective concentrations of the adenylate cyclase-activator forskolin and the cyclic AMP analog 8(4-chlorophenylthio) cyclic AMP were used instead of norepinephrine. Moreover, age-associated differences were abolished if the phosphodiesterase inhibitor isobutylmethylxanthine was present in addition to norepinephrine, or alone. (4) Study of the rates of dephosphorylation of troponin I, as initiated with the beta-adrenergic antagonist propranolol, showed no change in half-time as a function of age: this indicates no change in protein phosphatase activity. (5) These results suggest that there is less active net formation of cyclic-AMP in senescent heart cells in response to the neurotransmitter norepinephrine, giving a lesser activation of c-AMP-dependent protein kinase and less phosphorylation of these target proteins. PMID- 2561161 TI - Increase in plasma noradrenaline concentration after the administration of phentolamine in the patients with "latent" left-sided heart failure. AB - We measured increments of peripheral venous pressure induced by dynamic leg exercise (delta VP) in 10 healthy subjects (Group C) and 70 patients with heart diseases which primarily affect the left-side of the heart. None of the subjects showed apparent symptoms of left- or right-sided heart failure. The patients were divided into 2 groups on the basis of delta VP, namely, Group N (delta VP less than 35 mmH2O, n = 30, normal reaction) and Group H (delta VP greater than or equal to 35 mmH2O, n = 40, abnormal reaction). We measured the increments of plasma concentrations of noradrenaline (delta NAPH) and adrenaline (delta APH) with infusion of phentolamine (PH). Parallel studies with nitroglycerin and prazosin supplied strong evidence that delta NAPH was brought about mainly by the blockade of alpha 2-receptors at the sympathetic nerve terminals. Thus, we estimated the degree of sympathetic nerve activity from the central nervous system by opening using PH the negative feed-back loop for noradrenaline (NA) release at the sympathetic nerve terminals, and this degree of sympathetic nerve activity was compared with the degree of delta VP. The results obtained were 1) there was a rough overall correlation between delta VP and delta NAPH in the subjects of Groups C, N and H, and 2) delta NAPH was significantly higher in Group H than in Groups C and N. These results suggest that much reliance can be placed on the measured increment of plasma NA concentration in response to the administration of PH in assessing the degree of enhanced sympathetic nerve activity in the patients with "latent" left-sided heart failure. PMID- 2561162 TI - Cardiovascular responses to a newly developed phosphodiesterase inhibitor, DN 9693 [7-pyperidinyl-1,2,3,5-tetrahydroimidazo (2,1-b)-quinazolin-2-one.HCl] in dog cross-circulated atrial and ventricular preparations. AB - The cardiovascular effects of DN-9693, a newly synthesized cyclic AMP phosphodiesterase inhibitor, and sulmazole were investigated in an isolated right atrial or left ventricular preparation which was cross-perfused with blood from another support dog. Each substance dose-dependently increased heart rate and decreased blood pressure of the support dog when administered i.v. The hypotensive effects of DN-9693 were about 40 times more potent than those of sulmazole. The positive chronotropic effect of DN-9693 was about 10 times more potent, but the maximal increase in heart rate was much less than that produced by sulmazole. Each substance dose-dependently increased the sinus rate, the right atrial and the left ventricular developed tension in isolated preparations when administered selectively into cannulated arteries. The effects of DN-9693 were about 10-30 times more potent than those of sulmazole, although the maximal positive chronotropic response to DN-9693 was significantly smaller. Positive chrono- and inotropic effects induced by both substances in the isolated, blood perfused right atrial preparation were not inhibited by treatment with propranolol, but they were consistently attenuated by electrical stimulation of intramural vagal nerve fibers. These results suggest that DN-9693 and sulmazole produce cardiotonic effects by directly increasing the cyclic AMP level in the dog heart. DN-9693 possesses more potent cardiotonic and vasodilating effects than sulmazole, and has a smaller effect on heart rate. PMID- 2561163 TI - Complete atrioventricular block associated with regional myocardial scarring in a patient with Coxsackie B2 myocarditis. AB - Transient atrioventricular (A-V) conduction abnormalities are often experienced in patients with evolving acute viral myocarditis, but persistent complete A-V block requiring permanent cardiac pacing is rare. We describe a case who developed irreversible complete A-V block during the long-term course of Coxsackie B2 myocarditis. The endomyocardial biopsy revealed inflammatory cellular infiltrates and myocyte necrosis. A left ventriculogram and echocardiogram consistently demonstrated an aneurysm in the basal portion of the interventricular septum. It was speculated that the extensive myocardial scar caused by acute myocarditis resulted in the ventricular aneurysm of this particular myocardial region involving the A-V conduction system. PMID- 2561164 TI - Ivermectin, levamisole and thymic extract for chemotherapy and immunostimulation of visceral leishmaniasis in hamsters and mice. AB - The anti-leishmanial activity of ivermectin, pentostam or combination of pentostam with either levamisole or thymic extract was tested against Leishmania donovani infection in hamsters and mice. In vitro peritoneal macrophage-parasite interaction, the effect of splenic cells on the interaction of macrophages and parasites, spleen weight, parasite burden in spleen tissue as well as the antibody titers using micro-ELISA were used as parameters for evaluating the efficacy of these chemotherapeutic regimens. Treatment with ivermectin and immunoenhancement with pentostam combined with levamisole gave best results in both animal models. Furthermore, regimens used in this work were all active in reducing phagocytosis of promastigotes by macrophages in vitro. PMID- 2561165 TI - Unaffected electrogenic Na-K pump activity in "diseased" human atrial fibers, as assessed by intracellular K+ activity. AB - To investigate the role of the electrogenic Na-K pump in the resting membrane of "diseased" or "depolarized" human atrial muscles, intracellular K+ activity (aiK) and resting membrane potential (Vm) were simultaneously measured using double barreled K(+)-selective microelectrodes. Under perfusion with normal Tyrode's solution (37 degrees C) containing 5.4 mM [K]o, Vm averaged -43.9 +/- 1.4 mV, and aiK was 99.7 +/- 1.3 mM (mean +/- S.E., n = 33). The aiK was comparable to that of atrial muscles obtained from other intact mammalian species. In 5.4 mM [K]o, dihydro-ouabain (DHO) at concentrations of 10(-6) and 10(-5) M significantly decreased aiK and depolarized Vm. Similar decreases in aiK were observed when [K]o was decreased from 5.4 to 0.5 mM or when the temperature of the perfusing solution was decreased from 37 to 22 degrees C. Upon returning [K]o from 0.5 to 5.4 mM at 37 degrees C, aiK increased, Vm hyperpolarized markedly for about 3 min, and this was followed by less marked levels of hyperpolarization in the steady state. The high [K]o-induced increases in aiK were inhibited in the presence of DHO, and at low temperature (22 degrees C). Isoproterenol (10(-7) M) increased aiK and hyperpolarized Vm. Acetylcholine (10(-5) M) hyperpolarized Vm with no change in aiK. The rate of reduction of Na(+)-efflux during application of DHO (10(-5) M) was calculated based on the change in aiK and surface-to-volume ratio of the cell measured electronmicroscopically in the same tissue, and estimated to be 2.6 to 3.8 pmol/(cm2.s), close to the value reported for Purkinje fibers excised from intact animals. We conclude that the Na-K pump functions normally even in "diseased" human atrial muscles, thereby keeping aiK within a physiological range. PMID- 2561166 TI - [Effect of stimulation of adrenergic beta receptors on electrophysiologic properties of the bundle of Kent in patients with intermittent Wolff-Parkinson White syndrome]. AB - In 15 patients with intermittent ventricular preexcitation symptoms the effect of intravenously administered dobutamine (10 mcg/kg b.w./min) on the ecg tracing, effective atrial, a-v nodal and Kent bundle refractions as well as the incidence of cardiac arrhythmias were assessed basing on results of the electrophysiologic examination. Pharmacological stimulation of the adrenergic system resulted in highly statistically significant shortening of examined refraction periods (p less than 0,02 for effective Kent bundle refraction and p less than 0,001 for the rest parameters) and improved the conduction via the accessory pathway as well as exacerbated cardiac arrhythmias. PMID- 2561167 TI - [Effect of selective and non-selective adrenergic beta receptor blockaders on selected electrophysiologic properties of the human heart]. AB - Beta-2 adrenergic receptors were found in a human heart and their role in regulation of inotropic properties during last year was proved. The aim of our study was comparison of effects of selective (metoprolol) and nonselective (propranolol) beta-receptors blockade on cardiac electrophysiological properties. The study was carried out in 20 patients in the majority without organic heart injury and clinical symptoms of the sinus node dysfunction as well as atroioventricular conduction disorders. Method of the transoesophageal left atrial stimulation was utilized. Examinations were performed after drugs administrations in the same persons, during following days. Propranolol and metoprolol were intravenously administrated in a dose of 0.2 mg/kg b.w. Both drugs statistically significant lengthened sinus rhythm cycle time, sinus node recovery time, sinus node and a-v node effective refraction and lowered Wenckebach's point. They did not significantly effect on sinoatrial conduction and the atrial effective refraction. There were no significant differences between examined beta-blockers. Obtained results allowed us to conclude that: 1) eletrophysiologic properties of propranolol and metoprolol are similar, 2) it seems that in physiological conditions the effects of the adrenergic nervous system on electrophysiologic properties of sinus node, atrio-ventricular node and atrium is mainly realized by beta-1 adrenergic receptors. PMID- 2561168 TI - The effects of vasoactive drugs on halothane inhibition of contractions of rat mesenteric lymphatics. AB - The effects of halothane on alpha-adrenergic receptors, beta-adrenergic receptors, and the process of Ca++ dependent contractions on rat mesenteric lymphatics were examined. Halothane depressed the contraction rate of mesenteric lymphatics but did not effect the increase in lymphatic contraction rate caused by noradrenaline (an alpha-agonist). The depressant effect of halothane on the contraction rate was also not antagonized by propranolol (a beta-blocker), but was partly reversed by CaCl2. These findings suggest that the depressant effect of halothane on the lymphatic contraction rate derives not from blocking lymphatic alpha-receptors or stimulating lymphatic beta-receptors but rather by halothane inhibition on the process of Ca++ dependent lymphatic contractions. PMID- 2561169 TI - Recent advances on Epstein Barr virus and associated diseases. PMID- 2561170 TI - Susceptibility of non-fermentative gram-negative organisms to ureidopenicillins and quinolones. AB - The susceptibility of non-fermentative Gram-negative bacteria (excluding Pseudomonas aeruginosa) to ureidopenicillins and new quinolones was investigated. The ureidopenicillins were not active against the strains except against Pseudomonas species, 90% of which were inhibited by 64 mg/l. The new quinolones, particularly ciprofloxacin, had excellent activity (MIC range 0.06-8 mg/l) against these strains which included those resistant to beta-lactams and aminoglycosides. This suggested a promising alternative group of compounds to be used in chemotherapy of infections caused by non-fermenters. PMID- 2561171 TI - Measurement of intactness of rat liver endoplasmic reticulum. PMID- 2561172 TI - Transport of amino acids and selected anions in yeast. PMID- 2561173 TI - Isolation of everted plasma membrane vesicles from Neurospora crassa and measurement of transport function. PMID- 2561174 TI - Measurement of proton leakage across mitochondrial inner membranes and its relation to protonmotive force. PMID- 2561175 TI - Influence of calcium ions on mammalian intramitochondrial dehydrogenases. PMID- 2561176 TI - [Topoisomerase I from human placenta. Functional activity of products of expression of cloned cDNA fragments]. AB - Immunoscreening of the human placenta cDNA-library in the expression vector lambda gt11 using non-isotope detection based on the avidin-biotin system allowed to identify a number of clones encoding human topoisomerase I. The fusion protein from an extract of Escherichia coli cells infected with the recombinant phage lambda gt11 interacts with the monoclonal antibody raised against topoisomerase I from calf thymus; the dissociation constant being 5.7.10(-8) M. The restricted DNA fragments coding for the topoisomerase polypeptide in the composition of the fusion protein were recloned, and expression in the pEX vector was obtained. The functional analysis of the expression products has enabled localization of the epitope of binding the monoclonal antibody. It was demonstrated that the identified fusion protein can be applied for diagnosis of autoimmune diseases. PMID- 2561178 TI - [The effect of Z-conformation on enzymatic activity of restriction endonucleases]. AB - Recombinant plasmid pGC20 containing (GC)9-insert into SmaI site of pUC19 has been used to study the inhibition of cleavage by six restriction endonucleases; KpnI, SacI, EcoRI and also BamHI, XbaI and SalI, due to Z-DNA formation in negatively supercoiled plasmid. The recognition sites of these enzymes were located at different distances on both sides of the (CG)10-sequence. It was shown that the inhibition of the cleavage by KpnI, SacI and EcoRI was decreased in this series as fast as the distance between recognition site and B-Z junction was increased, and no inhibition of cleavage by EcoRI was found. However, such a correlation was not found in the series of BamHI, XbaI and SalI. In contrast with EcoRI the cleavage by SalI was inhibited completely. These results indicate the difference for "sensitivity" of restriction endonucleases to the structural perturbations of DNA associated with B-Z junctions. It seems to depend on features of the enzyme-substrate interaction mechanisms and also on recognition and flanking sequences of DNA. Consequently, experiments with the inhibition of the cleavage by any enzyme can not help to determine the dimension of the region of DNA with altered structure. PMID- 2561177 TI - [Synthetic DNA-bindings ligands containing reaction centers specific for AT- and GC-pairs in DNA]. AB - In the present communication design, synthesis and DNA binding activities of three bis-netropsins and two netropsin analogs containing two N propylpyrrolecarboxamide fragments linked covalently to peptides Gly-Gly-(analog I) and Val-Val-Val-Gly-Gly-(analog II) are reported. Each bis-netropsin consists of two netropsin-like fragments attached to peptides -Gly-Cys-Gly-NH2 (compound IIIa), H-Gly-Cys-Gly-Gly-Gly-(compound IV) or Gly-Cys-Sar-NH2 (compound IIIb) which are linked symmetrically via S-S bonds. Physico-chemical studies show that each bis-netropsin carries 6 AT-specific reaction centers and covers approximately 10 base pairs upon binding to poly(dA).poly(dT). This indicates that two netropsin-like fragments of the bis-netropsin molecule are implicated in specific interaction with DNA base pairs. The peptide fragments of bis-netropsins IIIa and IV form small beta-sheets containing two-GC-specific reaction centers. The DNase I cleavage patterns of bis-netropsin-DNA complexes visualized by high resolution gel electrophoresis show that the preferred binding sites for bis netropsins IIIa and IV are identical and contain two runs of three or more AT pairs separated by two GC pairs. Specificity determinants of netropsin analog II binding in the beta-associated dimeric form are identical to those of bis netropsin IIIa thereby indicating that there is a similarity in the structure of complexes formed by these ligands with DNA. In the monomeric form analog II exhibits binding specificity identical to that of analog I. Replacement of C terminal glycine residues by sarcosines in the peptide fragments of bis-netropsin IIIa leads to a decrease in the affinity of ligand for DNA. PMID- 2561179 TI - [Structural organization of kinetoplast DNA and its compactization in a model system in vitro]. AB - Studies on compactization and decompactization of the genome are of great importance for elucidation of structural mechanisms taking part in the regulation of gene activity. Kinetoplast DNA (kpDNA) is a convenient model for studies of compactization processes. KpDNA represents unique structure ("network"), consisting of catenated circular molecules of two types: minicircles (900 b.p.) and maxicircles (40 000 b.p.). The compactization process of kpDNA in vitro caused by interaction with synthetic peptide-dansylhydraside trivaline was studied. It was shown that at the initial stages the hairpins are observed on minicircles as if triple rings are being organized. The formation of hairpin is probably favoured by the presence in the minicircles of bent DNA, a specific nucleotide sequence causing rigid bending of the DNA helix. The hairpin does not make contact with the neighbouring DNA segment to form a triple ring, because the sizes of minicircles are too small. The minicircles compactization is finished with a complete collapse of the minicircles with the formation of rod-like structures. The catenation causes branching of rod-like structures. As a result of their intermolecular interaction, the branched rod-like structures become thicker. The process is completed with formation of the compact network, its diameter being 3-6 times smaller compared to the initial one. PMID- 2561180 TI - Non-enzymic activation of the fifth component of human complement, by oxygen radicals. Some properties of the activation product, C5b-like C5. AB - Purified human C5 was converted non-enzymically to an activated form as defined by its ability to participate in reactive lysis. This conversion occurred following exposure to systems that generate oxygen radicals, namely addition of H2O2 in the presence of ascorbic acid and iron or the addition of xanthine oxidase, acetaldehyde and iron. The conversion of C5 to a functionally active species was iron-dependent and inhibited by hydroxyl radical scavengers such as DMSO. The findings suggest that OH. is the active oxygen species that converts C5. The conversion product of C5, termed C5(H2O2), is C5b-like due to its ability to bind C6 and cause reactive lysis. C5(H2O2) is much more stable than C5b obtained by complement convertases. Although C5(H2O2) has lost the binding site of native C5 for C3b it can be cleaved by complement-derived convertases; the cleavage is, however, less efficient than in the case of native C5. The resulting cleavage product, which is C5a-like, is chemotactic although C5(H2O2) is not chemotactic. C5(H2O2) serves as a better substrate for plasma kallikrein than native C5, resulting in the generation of a C5a-like chemotactic product. These data indicate that oxygen radicals can bring about a conformational change in C5, causing it to behave as a functionally activated molecule of the complement system. This may have implications for the role of complement and its activation in the inflammatory response. PMID- 2561181 TI - Antifungal prophylaxis with itraconazole in prolonged neutropenia: correlation with plasma levels. AB - Seventy-two patients with haematological malignancies were treated prophylactically with itraconazole during remission induction therapy. The incidence of proven fungal infections was 18%, of which 12.5% were fatal. Aspergillus, Torulopsis and Candida proved to be major invasive pathogens. Plasma levels of itraconazole were monitored for all patients. The occurrence of fungal infection is significantly greater in the group where no therapeutic plasma levels were obtained during at least two weeks. There is a clear need to obtain quick information on itraconazole plasma levels in order to adapt dosage during prophylactic treatment in immunocompromised patients. The influence of itraconazole on liver function tests could not be separated from concomitant cytostatic or antibiotic treatment. No jaundice directly related to itraconazole could be found. During itraconazole prophylaxis a shift from classic pathogens such as Aspergillus and Candida, to Fusarium, Torulopsis and Mucor, may be seen. PMID- 2561182 TI - Efficacy of itraconazole in blastomycosis in a murine model and comparison with ketoconazole. AB - Mice were infected intranasally with B. dermatitidis yeasts, and after infection treated orally. Itraconazole 50-150 mg/kg/day was protective (prolonged survival) against lethal infection, although the infection was not sterilized. Itraconazole was approximately 3 times as potent as ketoconazole. These results suggest advantages for itraconazole therapy clinically. PMID- 2561183 TI - Oral, topical and parenteral antifungal treatment with itraconazole in normal and in immunocompromised animals. AB - Itraconazole was dissolved in polyethylene glycol for oral and topical treatment and in hydroxypropyl-beta-cyclodextrin for oral, topical or parenteral treatment. Topical and oral treatment was successful in microsporosis, trichophytosis, skin and vaginal candidosis, pityrosporosis and eye mycosis by Candida, Fusarium and Aspergillus. Vaginal candidosis could be cured with a one-day topical or oral treatment. The same results could not be obtained with any of the reference compounds (griseofulvin, terbinafine, ketoconazole or fluconazole) on a mg per kg body weight base, nor on a % concentration base. Antifungal levels were determined by bioassay: biologically active antifungal levels were present in plasma and vaginal fluid of rats, after one oral dose of 10 mg.kg-1, for at least 72 and 96 hours respectively. This was in good correlation with findings on prophylaxis of vaginal candidosis. Itraconazole was also successfully used, in normal animals and animals immunodepressed with various agents, in disseminated and systemic diseases: trichophytosis, sporotrichosis, histoplasmosis, candidosis, aspergillosis and cryptococcosis. Oral and parenteral treatment with itraconazole was compared in various models to oral and parenteral fluconazole and to parenteral amphotericin B. The outcome with itraconazole was better than with the other antifungals. Meningeal cryptococcosis responded very well to itraconazole. Combination therapy of itraconazole and fluconazole was not superior to treatment with itraconazole alone. No side-effects were observed in relation to itraconazole treatment. PMID- 2561184 TI - Biochemical approaches to selective antifungal activity. Focus on azole antifungals. AB - Azole antifungals (e.g. the imidazoles: miconazole, clotrimazole, bifonazole, imazalil, ketoconazole, and the triazoles: diniconazole, triadimenol, propiconazole, fluconazole and itraconazole) inhibit in fungal cells the 14 alpha demethylation of lanosterol or 24-methylenedihydrolanosterol. The consequent inhibition of ergosterol synthesis originates from binding of the unsubstituted nitrogen (N-3 or N-4) of their imidazole or triazole moiety to the heme iron and from binding of their N-1 substituent to the apoprotein of a cytochrome P-450 (P 450(14)DM) of the endoplasmic reticulum. Great differences in both potency and selectivity are found between the different azole antifungals. For example, after 16h of growth of Candida albicans in medium supplemented with [14C]-acetate and increasing concentrations of itraconazole, 100% inhibition of ergosterol synthesis is achieved at 3 x 10(-8) M. Complete inhibition of this synthesis by fluconazole is obtained at 10(-5) M only. The agrochemical imidazole derivative, imazalil, shows high selectivity, it has almost 80 and 98 times more affinity for the Candida P-450(s) than for those of the piglet testes microsomes and bovine adrenal mitochondria, respectively. However, the topically active imidazole antifungal, bifonazole, has the highest affinity for P-450(s) of the testicular microsomes. The triazole antifungal itraconazole inhibits at 10(-5) M the P-450 dependent aromatase by 17.9, whereas 50% inhibition of this enzyme is obtained at about 7.5 x 10(-6)M of the bistriazole derivative fluconazole. The overall results show that both the affinity for the fungal P-450(14)DM and the selectivity are determined by the nitrogen heterocycle and the hydrophobic N-1 substituent of the azole antifungals. The latter has certainly a greater impact. The presence of a triazole and a long hypdrophobic nonligating portion form the basis for itraconazole's potency and selectivity. PMID- 2561185 TI - Mode of action of itraconazole: morphological aspects. AB - The broad spectrum of antifungal activity of itraconazole is verified by morphologic criteria at the light and electron microscopical level. Yeast and fungal species examined are Candida albicans, Cryptococcus neoformans, Paracoccidioides brasiliensis, Sporothrix schenckii, Pityrosporum ovale, Trichophyton rubrum and Aspergillus fumigatus. Exposures of cultures of these yeasts and fungi to itraconazole results in dose- and time-dependent alterations which vary in nature and intensity from one species to another. The most striking gross morphological change is seen in the biphasic species and consists of the abolishment of morphogenic development of the blastospore into the hyphal forms (C. albicans and P. ovale) and of the hyphal into the yeast forms (P. brasiliensis). Furthermore, the outgrowth and development of inoculated A. fumigatus hyphae into sporulating vesicles is almost completely abolished. The above mentioned effects on morphogenesis are achieved in the 10(-10)-10(-7) M range. Except for P. ovale, the earliest ultrastructural changes after itraconazole consist of abnormalities at the plasma membrane, the cell wall and cytoplasmic vacuoles. They precede a marked increase in cell volume, defective cell division, abortive hyphal outgrowth and loss of cell viability. These changes are identical to those previously described after miconazole and ketoconazole treatment. However, when compared to the other available azole derivatives sharing the same basic mechanisms of action, itraconazole displays an exceptionally strong activity against A. fumigatus which can be morphologically translated by a potent necrotizing action on hyphae and inhibition of vesicle formation and sporulation. The in vitro effects of itraconazole are supported by data obtained from microscopic examinations of samples derived from patients with experimental animals infected with various fungal organisms. PMID- 2561186 TI - Toxicological profile and safety evaluation of antifungal azole derivatives. AB - For the development of new systemically acting, oral antifungal azoles, it is of key importance to compare them with ketoconazole, the first available drug in this therapeutic class. Ketoconazole is a major breakthrough although hepatic side-effects as well as interactions with mammalian steroids might rarely occur during prolonged treatment. The prediction of these side-effects is difficult but the potential to interact with mammalian cytochrome P-450 enzymes is considered to be important. Therefore, for the selection of itraconazole a multidisciplinary approach was applied to study this potential. The present paper deals with the toxicological profile of itraconazole and its safety evaluation. In addition, a further comparison with ketoconazole and also with fluconazole is provided, in so far sufficient information is available. For the liver as a potential target organ, the available data indicate that itraconazole is not a predictable hepatotoxic drug in man. The major endocrine targets for overdosing with antifungal azoles are the adrenal cortex and the gonads. Endocrine studies show that itraconazole is not bearing a potential to interfere with steroid hormones in patients, which is a major improvement when compared to ketoconazole. In rats, elevation of serum cholesterol is observed especially after chronic exposure to itraconazole. This species-specific phenomenon leads at toxic dose levels to secondary events, especially in the long-term toxicity studies. In man, including those with existing hypercholesterolemia, serum cholesterol is not adversely affected by itraconazole. In pregnant rats, ketoconazole was shown to be teratogenic at high, toxic doses. The same observation has been made for itraconazole and this also might be true for fluconazole.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561188 TI - Symposium: Itraconazole and its use in systemic mycoses. Antwerpen, June 22, 1989. Proceedings. PMID- 2561187 TI - The clinical pharmacokinetics of itraconazole: an overview. AB - Itraconazole (R 51211) is the prototype of a class of triazole antifungals characterized by a high lipophilicity. This property determines to a large extent the pharmacokinetics of itraconazole and differentiates it from the hydrophilic triazole antifungal fluconazole. The pharmacokinetics of itraconazole in man are characterized by a good oral absorption, an extensive tissue distribution with tissue concentrations many times higher than in plasma, a relatively long elimination half-life of about one day and a biotransformation into a large number of metabolites. One of them, hydroxy-itraconazole, is antifungally active and explains why antifungal plasma levels, when measured by bioassay, are about three times the itraconazole levels measured by a specific HPLC-method. Distribution studies have shown that therapeutically active levels of itraconazole are maintained much longer in some infected tissues than in plasma. For instance, active levels persist for four days in the vaginal epithelium after a one-day treatment and for 3 weeks in the stratum corneum of the skin after treatment has been stopped. Unlike fluconazole, itraconazole does not interfere with mammalian drug metabolizing enzymes, minimizing the risk of interaction with concomitantly administered drugs. These pharmacokinetic properties may contribute to the high efficacy and safety of itraconazole in patients with various mycotic infections. New pharmaceutical formulations are being explored in order to broaden the application field of itraconazole to intravenous and oral therapy of patients with malabsorption. PMID- 2561189 TI - The in-vitro antifungal spectrum of itraconazole. AB - The activity of itraconazole on 6113 fungal strains belonging to 252 species was evaluated in fluid media. The test medium was brain heart infusion broth for all fungi, except for Pityrosporum ovale, for which it was Dixon broth. Most of the human and animal pathogens and a large number of saprophytes were highly sensitive: dermatophytes, Candida, Cryptococcus, Torulopsis, Pityrosporum, Aspergillus, Penicillium, Sporothrix, dimorphic fungi, phaeohyphomycetes, agents of eufungal mycetoma, Entomophtorales and various others. The majority of Fusarium species and the Zygomycetes were poorly sensitive. Itraconazole was not only fungistatic at low concentrations, but also fungicidal for the tested organisms, such as Microsporum canis, Trichophyton mentagrophytes, Candida albicans, C. tropicalis, Aspergillus fumigatus, P. ovale and Cryptococcus neoformans with or without replenishment. Itraconazole was able to block the morphogenetic transformation of C. albicans from the yeast phase into the (pseudo)-mycelium phase. PMID- 2561190 TI - Prophylaxis of fungal infections with itraconazole during remission-induction therapy. AB - Deep fungal infections are an increasing problem in the treatment of acute leukemias and malignant lymphomas. Risk factors are known but unavoidable. Because of diagnostic difficulties most patients are treated empirically with intravenous amphotericin B. This drug's toxicity increases morbidity and mortality. An orally absorbable triazole derivative, itraconazole, may offer effective and safe prophylaxis against deep candidosis and aspergillosis in these patients. Such infections have been treated successfully with oral itraconazole even when resistant to intravenous amphotericin B. In retrospective comparative studies there are significantly less deep fungal infections in patients given itraconazole. The significance of the difference varies between studies. Pharmacokinetic data confirm therapeutic plasma levels of itraconazole but with wide variation within and between patients. The current large, multi-centre, randomised, double-blind, prospective trial of oral itraconazole versus placebo in the U.K. will test its prophylactic efficacy against deep fungal infections during treatment of haematological malignancies. PMID- 2561191 TI - [Treatment of hyperkalemia using salbutamol in acute kidney failure]. PMID- 2561192 TI - Cytomegalovirus vasculitis and colon perforation in a patient with the acquired immunodeficiency syndrome. AB - We describe a patient with the acquired immunodeficiency syndrome who suffered a colon perforation which we believe was directly attributable to disseminated cytomegalovirus (CMV) infection. Thrombosed vessels within the submucosa and muscle wall contained evidence of CMV vasculitis, while adjacent vessels without viral inclusions were fully patent. This report supports other evidence that CMV may act as a primary etiologic agent of gastrointestinal disease, particularly in the immunocompromised host. The increased recognition of CMV as a cause of significant morbidity in certain gastrointestinal lesions becomes especially important with the advent of newer antiviral therapy specifically directed against CMV infection. PMID- 2561193 TI - [Contrast material for CT of the liver and spleen--an experimental study on the safety of lipiodol-iopamidol emulsion]. AB - Iodinated oil (Lipiodol Ultra-Fluid) emulsified by non-ionic contrast material (Iopamidol) featuring much iodine and surface activity, was developed for computed tomography (CT) of the liver and spleen. The stability study revealed that Lipiodol-Iopamidol Emulsion (LIE, 1-6 microns in diameter) stored 2 weeks under refrigeration showed no visible change in globule size distribution. The biodistribution study in six rabbits revealed that LIE injected into the vein was accumulated in the liver, spleen, and lung, which have reticuloendothelial cells, however not accumulated in the brain, kidney, and serum, which have no reticuloendothelial cells. The disruption to the blood brain barrier (BBB) was assessed using Evans' Blue dye as a visual marker. After the coarse emulsion (larger than 8 microns in diameter) was injected into the internal carotid artery of three rats, the permeability of BBB increased. However, after the injection of LIE, no increased permeability of BBB was noted. Our results suggest that LIE will have a clinical application in the detection of focal hepatic and splenic lesions. PMID- 2561194 TI - Hydroxylapatite coated dental implants. PMID- 2561195 TI - [Biosynthesis and transport of phospholipids in yeast cells]. PMID- 2561196 TI - Effect of oxiracetam on cerebral cholinergic imbalance secondary to an NMDA receptor blockade. PMID- 2561197 TI - Age-related increase in CGRP binding site densities in rat cerebellum. PMID- 2561198 TI - Stress-induced decrease of central benzodiazepine receptors number is not due to internalization. PMID- 2561199 TI - Human platelet activation by PAF: receptor characterization. PMID- 2561200 TI - Pharmacological characterization of glutamate non-NMDA receptor subtypes in hemisected baby rat spinal cord. PMID- 2561201 TI - Structure-activity relationship for steroid action on voltage-dependent characteristics of sodium and potassium channels. PMID- 2561202 TI - Effects of TRH on the immune system of newborn rats. PMID- 2561203 TI - Stereospecific effects of tocainide and its chiral derivatives on rat skeletal muscle sodium channel. PMID- 2561204 TI - Effect of doxorubicin on calcium receptors of myocardial cells. PMID- 2561206 TI - Release-regulating GABAB autoreceptors in rat cerebral cortex slices. PMID- 2561205 TI - Cross-talk between phospholipase C- and adenylate cyclase-coupled receptors in the rat hippocampus. PMID- 2561207 TI - Metastatic diversity in human prostatic carcinoma: implications of growth factors and growth factor receptors for the metastatic phenotype. PMID- 2561208 TI - Tn917 transposition in Lactobacillus plantarum using the highly temperature sensitive plasmid pTV1Ts as a vector. AB - pTV1Ts, a temperature-sensitive plasmid coding for chloramphenicol (Cm) resistance and carrying the macrolide-lincosamide-steptogramin B (MLS) resistance transposon Tn917, was introduced into strains of Lactobacillus plantarum by electroporation. After two passages in broth medium selecting for MLS resistance at 40 degrees C and subsequent plating on solid medium, two strains, L. plantarum NC4Ts1 and L. plantarum NC7Ts5, lost chloramphenicol resistance but retained MLS resistance, indicative of Tn917 transposition into host DNA. Analysis of DNA from MLSrCms isolates from both strains revealed Tn917 insertions into resident plasmids. Restriction analysis of plasmid DNA from four MLSrCms isolates from NC7Ts5 indicated four different insertion sites. PMID- 2561209 TI - Site-specific recombinations between direct and inverted res sites of Tn2501. AB - The resolvase gene and the putative res site of Tn2501 are not closely related to any of the previously described resolution functions. In view of this divergence, we designed genetic experiments to confirm the localization of the res site. We analyzed the activity of the Tn2501-encoded resolvase on substrates containing either directly or invertedly repeated res sites. These experiments confirm the localization of the res site that was predicted from nucleotide sequence data and show that the Tn2501 resolvase promotes site-specific inversions in vivo. PMID- 2561210 TI - Tn4527, a Tp Sp/Sm transposon related to Tn7 and flanked by IS1. AB - Tn4527 was isolated from a Salmonella typhimurium strain obtained in Brazil. Its size is 19.6 kb and it carries resistance to trimethoprim, spectinomycin, and streptomycin, as in the case of Tn7 (14 kb). A restriction analysis of the transposon shows regions of similarity to Tn7 mixed with extra DNA. The 2.6-kb and 2.2-kb HindIII fragments of Tn7, which encode transposition-related proteins, show homology to Tn4527. In contrast to Tn7, Tn4527 is flanked by direct repeats, which seem to be IS1's, as they have appropriate restriction sites and hybridize both to IS1 and to internal IS1 oligonucleotides. PMID- 2561211 TI - A 10- rather than 9-bp duplication associated with insertion of Tn5 in Escherichia coli K-12. AB - The composite transposable element Tn5, which is made up of two inverted IS50 elements surrounding genes encoding drug resistance, generally generates 9-bp duplications at the site of insertion. In our studies of three Tn5 insertion mutants at one location in the Escherichia coli chromosome, we have observed that one contains a duplication of 10 bp, while the other two have the usual 9-bp duplication. Three other insertion elements, IS1, IS4, and IS186, give variable sized target site sequence duplications. We observed a similarity of amino acid sequence in a small region of the putative transposases among IS4, IS186, and Tn5 suggesting a conservation of function in this group of transposases. PMID- 2561212 TI - Analysis of relative reversion frequencies for IS2 insertion mutations in the regulatory region of the galOPETK operon of Escherichia coli. AB - Two previously characterized mutations in the galOPETK operon of Escherichia coli, galOP-3 and galOPE-490, contain IS2 insertions only 1 bp apart in the gal regulatory region; yet only the former yields Gal+ phenotypic revertants at a detectable frequency. We have shown that the galOPE-490 allele comprises two mutations--an IS2(I) insertion at bp+(2-6) (relative to the gal mRNA start site) plus a C/G to A/T transversion at bp + 59. The latter creates an ochre stop codon and lies within the internal site of the bipartite gal operator; it acts as an operator mutation in an in vivo repressor titration assay. Analysis of a newly isolated allele (galOP-490*) which retains the IS2 of galOPE-490 but is galE+ reveals a reversion frequency approximately 30-fold higher than that of galOP-3. Reversion of galOPE-490 is at least 10,000-fold lower and has not been detectable even under conditions conducive to enhanced double mutations in other systems. PMID- 2561213 TI - Identification and nucleotide sequence of the minimal replicon of the low-copy number plasmid pBS2. AB - The plasmid pBS2 has a low copy number and is endogenous to Bacillus subtilis. The replication of this plasmid depends on the function of most of the host's dna genes including dnaB, which is unique to B. subtilis and is required for both the initiation of chromosome replication and the DNA-membrane association. We have identified the region that is essential for the replication of pBS2 and determined the complete 2279-bp nucleotide sequence of this region. In this region, there are two stretches of sequence homologous to the 18-bp consensus sequence which commonly appears at the origin of replication of plasmids pUB110 and pC194. The entire region contains six sizable open reading frames. Two of them are probably translated. One open reading frame, designated ORF A, coding for 269 amino acids, has significant homology, in terms of amino acid sequence, with the open reading frame of the gene for the Rep U protein of plasmid pUB110. The similarities between pBS2 and other plasmids suggest that the pBS2 may also replicate as a rolling circle, which appears to be the salient feature of a mechanism of replication that is common to small plasmids in gram-positive bacteria. PMID- 2561214 TI - SC-41930: an inhibitor of leukotriene B4-stimulated human neutrophil functions. AB - SC-41930 was evaluated for effects on human neutrophil chemotaxis and degranulation. At concentrations up to 100 microM, SC-41930 alone exhibited no effect on neutrophil migration, but dose-dependently inhibited neutrophil chemotaxis induced by leukotriene B4 (LTB4) in a modified Boyden chamber. Concentrations of SC-41930 from 0.3 microM to 3 microM competitively inhibited LTB4-induced chemotaxis with a pA2 value of 6.35. While inactive at 10 microM against C5a-induced chemotaxis, SC-41930 inhibited N-formyl-methionyl-leucyl phenylalanine (fMLP)-induced chemotaxis, with 10 times less potency than against LTB4-induced chemotaxis. SC-41930 inhibited [3H]LTB4 and [3H]fMLP binding to their receptor sites on human neutrophils with KD values of 0.2 microM and 2 microM, respectively. SC-41930 also inhibited neutrophil chemotaxis induced by 20 OH LTB or 12(R)-HETE. At concentrations up to 10 microM, SC-41930 alone did not cause neutrophil degranulation, but inhibited LTB4-induced degranulation in a noncompetitive manner. SC-41930 also inhibited fMLP- or C5a-induced degranulation, but was about 8 and 10 times less effective for fMLP and C5a, respectively. The results indicate that SC-41930 is a human neutrophil LTB4 receptor antagonist with greater specificity for LTB4 than for fMLP or C5a receptors. PMID- 2561215 TI - Extracardiac intrathoracic abnormalities demonstrated by Tc-99m RBC gated blood pool imaging. AB - In addition to intrathoracic great vessel abnormality, pericardial effusion, and splenomegaly, extracardiac intrathoracic abnormalities were found in 12 of 210 patients' 99mTc red blood cell (RBC) gated cardiac blood pool imagings. These abnormalities, including five cases of absent pulmonary perfusion due to tumor mass, four of pleural effusion, two of pneumothorax, and one of left lung mass attenuation, were confirmed with concurrent or subsequent chest radiography, chest CTs, or biopsy. Pulmonary blood pool activity is normally seen on both sides in both anterior and left anterior oblique views; decreased or absent perfusion on either side or in part of the lung may indicate chest/pulmonary pathologies. Although pulmonary and thoracic wall lesions are not frequently seen, such incidental findings during gated cardiac blood pool imagings can lead to further study for these clinically unsuspected lesions and may benefit the patient. PMID- 2561216 TI - Increased iodine-123-iodoamphetamine uptake in hepatomas. AB - In the current study, two cases of hepatoma are reported in which N-isopropyl-(I 123)-p-iodoamphetamine (IMP) liver scan demonstrated increased accumulation in the tumor corresponding to the areas enhanced on contrast enhanced CT (CE-CT). In contrast, there was no IMP accumulation in the necrotic area of the tumor in which no enhancement was found on CE-CT. Thus, IMP liver scan seems to have the potential to assess the viability of a hepatoma as well as to detect and localize it. PMID- 2561217 TI - [Granular cell bronchial tumor associated with acoustic neurinoma. Apropos of a case]. PMID- 2561218 TI - [Synchronous small cell bronchial cancers]. PMID- 2561219 TI - Electron microscopy of lipid deposits in human atherosclerosis. AB - The filipin probe associated with tannic acid stain was used to study intra- and extracellular lipids in surgically removed human atherosclerotic lesions (n = 20). In particular, intimal thickenings, fatty streaks and fibrolipidic plaques have been investigated by using mainly transmission and scanning electron microscopy. In the intimal thickenings, the lipid deposits were mainly localized in the subendothelial space as homogeneously sized particles (40-140 nm) and more heterogeneous uni-multilamellar vesicles (35-700 nm). Intermediate lipid forms were also observed. In the fatty streaks, the lipid deposits were intracellular and mainly observed in cells with a monocyte/macrophagic phenotype. Lipid inclusions, lipid lysosomal bodies and intracellular cholesterol crystals very similar to those observed in experimentally induced atherosclerosis were documented. In the fibrolipidic plaque the lipid deposits were found both in the intracellular and in the extracellular compartments. Lipids accumulated within arterial macrophages and smooth muscle cells, usually as lipid droplets. Clusters of lipoprotein-like particles (50 nm in diameter) as well as larger uni multilamellar lipids (700 nm) with an occasional compound appearance were particularly observed bound to elastic tissue and collagen fibers. These morphological observations outline the complexity of lipid metabolism in the various histological aspects of human atherosclerosis. PMID- 2561220 TI - Non-fibrous inorganic particles in human bronchoalveolar lavage fluids. AB - Bronchoalveolar lavage (BAL) is a simple and non-invasive sampling technique of the deep lung. Analytical electron microscopy was used for the identification and quantification of non-fibrous inorganic particles recovered in BAL fluid samples from 51 subjects with various occupational exposures (silica, silicates, metals and alloys, metallic oxides, precious and hard metals, abrasives). Around 4750 particles were analysed. More than sixty different compounds were identified, among which silica, kaolinite, illite, mica, Fe oxides and hydroxides, appeared to be ubiquitous. Feldspar, talc, chlorite, Al oxide, Ti oxide, tungsten carbide, stainless steel, carbonaceous compounds and flyash were also frequently encountered. From 1 to 21 compounds were identified in each sample. Repeated BAL samples obtained for 2 subjects did not show significant differences. Particles characteristic of the occupational exposure were found in BAL up to 21 years after cessation. BAL content can also reflect mixed occupational exposures. Absolute particle concentrations measured in twelve samples ranged between 0.1 and 9.9 x 10(6) particles/ml BAL fluid and mean particle diameter ranged between 0.5 and 1.2 microns. Mineralogical analysis of non-fibrous particles in BAL can be a useful tool to investigate occupational exposures. It allows, in most cases, a better characterization of the exposure than medical questioning. It may be helpful in identifying pathogenic particles, however it must be kept in mind that a positive result is only a proof of exposure and never a proof of disease. The main limitations of this technique are difficulties in sampling severely diseased subjects and inaccuracy in detecting easily soluble compounds and particles with a high rate of alveolar clearance. PMID- 2561221 TI - [Electrical cell-to-cell coupling in the myocardium]. PMID- 2561222 TI - [The cellular mechanisms of hypoxic pulmonary vasoconstriction]. PMID- 2561223 TI - [Neuropeptide antagonists]. PMID- 2561224 TI - [The voltage dependency calcium channels]. PMID- 2561225 TI - [Protein kinase C and cellular regulation]. PMID- 2561226 TI - [Antiopiate effects of cholecystokinin octapeptide in the regulation of pain sensation and an analysis of its mechanism]. PMID- 2561227 TI - [Selective inhibitory effects of flufenamic acid on metabolic pathways of arachidonate 5-lipoxygenase and cyclooxygenase]. PMID- 2561228 TI - [The effects of calcium antagonists on the nervous system]. PMID- 2561229 TI - [Insulinoma in a female diabetic]. PMID- 2561230 TI - [The level of R proteins in reactive arthritis]. AB - A total of 46 patients with reactive arthritides and 20 patients with rheumatoid arthritis (RA) were examined, as compared with 20 normal persons, for the blood serum content of the products of catabolic breakdown of cell receptors--R proteins (according to inhibition of the hemagglutination reaction between human red blood cells and anti-R-serum) and for conventional parameters of phagocyte function of leukocytes. The content of R proteins in patients with reactive arthritis and RA was found to be increased, and the phagocytic activity was discovered to be lowered. The changes in the parameters indicated depended on the disease entity and the disease course. The possible pathophysiological importance of changes in the level of R proteins in arthritides is discussed. PMID- 2561231 TI - [Cytomegalovirus infection in clinical medicine]. AB - The author provides the data on studies into cytomegaloviral infection for many years, the results of examinations made during prospective clinico-virological and immunological follow up of 236 women with an unfavourable obstetrical disease history and verified cytomegaloviral infection. The diagnosis, epidemiology and the spectrum of the clinico-pathogenetic patterns of the infection are under discussion. The use of the immunocorrective drugs (levamisole, T activin) for the treatment of the private patterns of cytomegaloviral infection provided beneficial results. The prospects of further study into the problem are analyzed. PMID- 2561232 TI - [The role of Coxsackie B viruses in the etiology of dilated cardiomyopathy]. AB - In order to establish the relationship between Coxsackie B virus infection and dilated cardiomyopathy (DCMP), 53 patients suffering from DCMP underwent serological tests. The rate of demonstration of virus-neutralizing antibodies at different titres was compared with the respective values in the previously examined 180 patients with myocarditis, 58 patients with myocardial sclerosis and 150 normal persons. In DCMP, antibodies against B2 type virus were detectable more frequently (p less than 0.001) and at higher titres (p less than 0.05) that in normal persons. Antibodies at the titres greater than 1:128 were most demonstrable in myocarditis and then at an equal rate in DCMP and focal myocardial sclerosis following Coxsackie myocarditis. The conclusion is drawn about the relationship between Coxsackie infection and DCMP as well as between DCMP and myocarditis. The reported data on the mechanisms by which myocarditis progresses to DCMP are discussed. PMID- 2561233 TI - Anticoagulant and antithrombotic properties of synthetic sulfated bis-lactobionic acid amides. AB - Sulfated bis-lactobionic acid amides, a new class of polyanions with heparin-like properties, were synthesized and their antithrombotic and anticoagulant activities were determined. Compared to heparin and Fragmin, a low molecular weight heparin, the substances exhibited moderate to low anticoagulant activities in aPTT, thrombin clotting time and Heptest assays. In amidolytic assays no anti IIa activity and only exceedingly low anti-Xa activity was observed. The antithrombotic activity of the bis-lactobionic acid amides was determined using two thrombosis models. In rabbit and rat models thrombi were induced by a combination of endothelial damage and reduction of blood flow or only by endothelial damage. At least one of the bis-lactobionic acid amides (LW 10082) exhibited a considerable antithrombotic activity which was similar to low molecular weight heparin. PMID- 2561234 TI - Mechanisms of beta-adrenoceptor mediated increase in delayed rectifier potassium current. AB - Experiments were carried out in single ventricular cells of the guinea-pig heart. Isoproterenol, forskolin, intracellularly applied cyclic AMP and 3-isobutyl-1 methylxanthine increased the delayed rectifier potassium current (IK). The effect of isoproterenol was abolished by intracellularly applied guanosine 5'-O-(3-thio triphosphate). These results indicate that isoproterenol stimulates beta adrenoceptors to activate adenylate cyclase by mediation through the stimulatory GTP-binding protein, and causes an increase in intracellular cyclic AMP levels. Then IK is probably increased by phosphorylation of the IK-channel protein by cyclic AMP-dependent protein kinase. PMID- 2561235 TI - [Diagnosis and treatment of hygroma in children]. PMID- 2561236 TI - [Experimental infection caused by Machupo virus in intact and immunosuppressed albino mice]. AB - Features of the course of experimental infection in white mice caused by Machupo virus in relation to their age, route of virus inoculation, and under conditions of normal and suppressed immune response created by antithymocyte globulin (ATG) were studied. The effect of different ATG doses and intervals of its administration on the experimental infection were demonstrated. PMID- 2561237 TI - [Comparative study of antiherpetic effect of 9-beta-D-xylofuranosyl- adenine and 9-beta-D-arabinofuranosyladenine and their inhibition of defective herpes simplex type 1 virus in a cell culture]. PMID- 2561238 TI - [Increased effectiveness of electron microscope diagnosis of orthopoxvirus infections]. PMID- 2561239 TI - [Immunoenzyme analysis in screening for antibodies against herpes simplex virus]. PMID- 2561240 TI - [Filtration-chromatographic method of purification of rotaviruses]. PMID- 2561241 TI - [Systemic photochemotherapy (PUVA) in acanthosis nigricans maligna: regression of keratosis, hyperpigmentation and pruritus]. AB - We report on a 60-year-old patient, who developed malignant acanthosis nigricans (MAN) with intense itching 2 years after a large-cell bronchial carcinoma had been diagnosed and found inoperable. The MAN became manifest at a phase of full clinical remission of the lung tumor, which had been treated with cytostasis (cisplatin, vindesine), high energy irradiation, and extirpation of the lymph node metastases. One year after onset of MAN, the lung tumor relapsed, accompanied by an elevated serum level of carcinoembryonic antigen (CEA). The patient was slightly obese, but not diabetic. The generalized MAN was treated with 18 exposures to systemic PUVA (photochemotherapy) over 9 weeks. The patient received 8-methoxypsoralene (8-MOP) orally and a total UVA dose of 52 J/cm2; the last exposure amounted to a maximum dose of 4 J/cm2. Under this treatment, the patient was completely relieved from tormenting pruritus; in addition, we observed significant regression of the pigmented keratoses as well as the intertriginous maceration. PMID- 2561242 TI - [50th anniversary of the diagnosis of Japanese encephalitis in the Primorsk territory]. PMID- 2561243 TI - Luminol dependent chemiluminescence of quartz and zymosan stimulated neutrophils. AB - The effect of the opsonization by zymosan and quartz particles on the chemiluminescence was investigated on human neutrophil granulocytes. Opsonization of zymosan enhanced the chemiluminescence response, while opsonized quartz inhibited the chemiluminescence reaction. Calcium ionophore A 23187 treatment did not influence the chemiluminescence of quartz but the light signal in the presence of quartz decreased rapidly. In parallel experiments the protein pattern of zymosan treated neutrophils was investigated by high resolution two dimensional polyacrylamide gel electrophoresis. PMID- 2561244 TI - Role, mechanism of action and application of gonadoliberins in reproductive processes. AB - Gonadoliberin (gonadotropin releasing hormone, GnRH) plays a central role in the regulation of reproductive functions as it regulates the release of both luteinizing hormone (LH) and follicle stimulating hormone (FSH). The isolation and structure determination of GnRH opened the possibility of its use for influencing reproductive processes. This possibility initiated a rapid development in the design of potent and long-acting GnRH agonists and antagonists. The most important structural modifications of GnRH leading to superagonists are the D-amino acid substitutions in position 6 combined with Pro9 ethylamide or azaGly10 at the C-terminus. We have synthesized several superagonists of GnRH according to these substitution principles. Furthermore, our L-isoaspartyl modification in position 6, as a new approach to GnRH agonist design, also resulted in superactive analogs. The recently discovered sequences of non-mammalian GnRH-s opened new routes for us to synthesize species specific GnRH agonists. All three groups of the above mentioned GnRH analogs have been successfully used for the treatment of sexual disorders of different animals (cattle, pigs, rabbits, etc.). Ovulation synchronization and a 30% increase in the fertility rate could be achieved by using GnRH agonists in cattle breeding. Analogs derived from species specific sequences could be applied for the induced artificial propagation of fish even out of the spawning season. It is known that superactive GnRH analogs can suppress the growth of certain hormone-dependent tumours. In vitro and in vivo tests of our analogs showed promising antitumour activity in breast cancer which might be explained by the mechanism of desensitization. Almost a hundred antagonist analogs of GnRH have been developed in our laboratory. The most effective ones contain 4 or 5 D-amino acids, and one of them is even orally active. The inhibition of ovulation can also be achieved by the administration of GnRH superagonists. This phenomenon might also be explained by the desensitization of LH-release. Radioactive analogs specifically labeled with tritium in different amino acid residues have been synthesized and used for studying tissue distribution and biodegradation of gonadoliberins. Analogs containing a photoreactive group have been prepared and applied for the trials of GnRH receptor isolation. PMID- 2561245 TI - Cell-mediated transmission of hormonal imprinting to virgin mammalian cells in culture and failure of transmission with the nutrient medium. AB - Chinese hamster ovary (CHO) cells and Chang liver cells which had already interacted with a hormone (gonadotropin, TSH, insulin) in culture, transmitted hormonal imprinting to virgin cells not previously involved in the interaction. The information associated with imprinting was not mediated by the nutrient medium, because the nutrient medium of the hormone-treated cells did not induce imprinting in virgin cells and even reduced rather than enhanced the hormone binding capacity thereof. Thus the transmission of information is in all probability associated with a direct cell-cell contact. PMID- 2561246 TI - Effects of ischemia on opioid receptors in newborn pig lumbar spinal cord. AB - The characteristics of opioid receptors were studied by the binding of (3H)naloxone in ischemic lumbar spinal cord segments of newborn pigs. Ischemia was elicited by ligating the aorta for 30 min. The number of millimicron opioid receptors decreased, from 117 +/- 18 to 89 +/- 11 fmol/mg protein, while the Kd value was not significantly altered. It is concluded that even a relatively brief interruption of the oxygen supply may cause severe damage in the lumbar spinal cord of the newborn pig, affecting the opioid neurotransmission. The animal model employed here might be suitable for studying the effects of hypoxia in newborns and children during chest operations involving the descending aorta. PMID- 2561247 TI - Folate coenzyme and antifolate transport proteins in normal and neoplastic cells. AB - The transport systems for folate coenzymes and antifolate compounds into various types of normal and neoplastic cells display considerable diversity in their pharmacokinetics and also in terms of the apparent molecular weights of the proteins involved. Further, several uptake routes may exist in a given cell type. A variety of neoplastic tissues have been reported to rely upon a single major transport system that has a relatively high affinity for the reduced form of folate compounds and for antifolates such as methotrexate. Using a photoaffinity analogue of methotrexate, we have identified the involvement of a 48 kDa membrane protein and a 38 kDa cytosolic or peripheral membrane protein in the transport of this compound into murine L1210 leukemia cells. Such an uptake is absent in mutant L1210 cells that are defective in methotrexate transport. We propose a model for the uptake of reduced folate coenzymes in L1210 cells in which the compound is initially transported across the cell membrane by the 48 kDa protein and delivered on the cytoplasmic surface to the 38 kDa protein; the 38 kDa protein then carries the folate compound to a specific enzyme of folate metabolism. Antibodies to the membrane folate binding protein from human placenta cross-react with the 48 kDa protein in L1210 cell membranes indicating an immunological relationship between these two proteins. Comparison of the amino acid sequences of peptides of the placental receptor obtained by digestion with S. aureus V8 protease indicate the presence of two homologous forms of the folate binding protein in placenta; one of these forms appears to have an identical sequence to the soluble and membrane associated folate binding proteins in human epidermoid carcinoma (KB) cells, which in turn share the primary structure of the soluble and membrane associated folate binders in human milk in the regions that have been sequenced. These results indicate that the folate coenzyme transport proteins in various tissues may be structurally related and in several instances may even be identical. In the latter cases the observed differences in apparent molecular weights may be due to differences in glycosylation and/or proteolysis. In support of this view is our observation that the deglycosylated and/or partially proteolyzed placental receptor retains the ability to bind folate. PMID- 2561248 TI - Studies on drug resistance in chronic lymphocytic leukemia. AB - Chronic lymphocytic leukemia is a neoplastic disease in which drug resistance invariably occurs. We have studied the uptake and interaction with molecular targets of two drugs, chlorambucil and adriamycin, in CLL lymphocytes and CHO cell lines. Resistance does not appear related to uptake for either drug. Exposure to CLB causes DNA cross-links in the sensitive but not in the resistant cell line. The GSH content of B-CLL lymphocytes is depleted after a 20-hr incubation. An inability to maintain its GSH content may contribute to this cell's vulnerability to CLB. The resistance of CLL lymphocytes to ADR may be related to the undetectable levels of its target enzyme DNA topoisomerase II. Future approaches may involve study of novel anthracyclines, DNA topoisomerase I inhibitors and the development of in vitro predictive tests. PMID- 2561249 TI - Mediation of cellular anion detoxification in leukemic cells by unidirectional efflux pumps. AB - Methotrexate influx in L1210 cells is mediated almost entirely by a single system, whereas efflux occurs via three routes, the influx carrier functioning in reverse and two additional systems which are operationally unidirectional. The two unidirectional routes show considerable similarities in energy-dependence and inhibitor sensitivities but can be separated by their differential inhibition by bromosulfophthalein (BSP), probenecid, and vincristine. The predominant route is inhibited by BSP and vincristine, whereas the other route is sensitive to probenecid. A search for additional anion substrates for the unidirectional efflux systems for methotrexate led to the finding that cholate exits L1210 cells via the same two unidirectional efflux systems as methotrexate. Cholate efflux is carrier-mediated, energy-dependent, and unidirectional, and it can be inhibited by several compounds which inhibit the efflux of methotrexate. Moreover, the same concentration of each compound which produced a half-maximal inhibition of methotrexate efflux also inhibited the efflux of cholate by 50%. Cholate efflux occurred predominantly (85%) via the BSP-sensitive route. The observation that methotrexate and cholate share the same efflux systems in L1210 cells suggest that perhaps other large anions are also accommodated by these systems since methotrexate and cholate differ in net negative charge and have few common structural features. The hypothesis advanced from these studies has been that the function of the unidirectional efflux systems is to extrude various organic anion catabolites which might otherwise become toxic if allowed to accumulate within cells. Possible intracellular anions in this latter category include sulfonated or carboxylated steroids, bilirubin, and products of vitamin D and prostaglandin catabolism. Unidirectional efflux pumps have been identified for other anionic compounds including cyclic AMP and oxidized glutathione, and in both cases similarities were apparent with the efflux of methotrexate and cholate. The most compelling comparison was with prostaglandin A1 which inhibited the efflux of cyclic AMP, methotrexate, and cholate half-maximally at the same low concentration of 0.1 microM. Energy-dependent unidirectional efflux pumps for large neutral or cationic drugs have also been identified in cells with acquired multidrug resistance. The latter efflux activity is thought to be mediated by a membrane glycoprotein (p170) which is also sensitive to several of the various inhibitors (reserpine, verapamil, and quinidine) which reduce the efflux of methotrexate and cholate. PMID- 2561250 TI - Singular point, organizing center and acupuncture point. AB - A hypothesis is proposed on the nature of acupuncture point and organizing center, the role of meridian system in growth regulation, and the mechanism of acupuncture. Both organizing centers and acupuncture points have low electric resistance. The low electric resistance is related to the distribution of gap junction and thus intercellular communication. Some acupuncture points may be organizing centers. The meridian system is important in coordination and regulation of morphogenesis. The properties of organizing centers and acupuncture points can be explained in view of singular point. Coupling and oscillation may underlie the mechanism of acupuncture as well as growth regulation. PMID- 2561251 TI - Primary tumors of the exocrine pancreas. Classification, overview, and recent contributions by immunohistochemistry and electron microscopy. AB - Histologic classification and subtyping of primary tumors of the exocrine pancreas have great prognostic value, although most tumors--such as duct cell adenocarcinomas--are consistently high grade with respect to biologic behavior. The mortality rate for these tumors is nearly equal to the incidence rate, and prognosis does not appear to be significantly influenced by histologic grade of the primary tumor. Other tumors, such as solid and papillary epithelial neoplasms, have intermediate to low malignant potential, while microcystic adenomas have proved to be essentially benign. Recent application of immunohistochemistry, along with electron microscopy, has helped to elucidate the histogenesis of some tumor subtypes. An intestinal phenotype has been demonstrated for mucinous cystic neoplasms with goblet cells, Paneth cells, and a variety of endocrine cells. Divergent tumor differentiation has been seen in a rare group of mixed tumors with variable composition of duct, acinar, or endocrine cells. Some infantile pancreatic tumors, known as pancreatoblastomas, may also show mixed-cell composition. Immunohistologic studies of solid and papillary epithelial neoplasms have not yet identified a consistent or reliable phenotype or marker, and ultrastructural studies have reached disparate conclusions regarding histogenesis. The rarity of other tumors--e.g., those with osteoclasttype giant cells--has hampered complete characterization. PMID- 2561252 TI - Benzodiazepine receptor-mediated enhancement and inhibition of taste reactivity, food choice, and intake. PMID- 2561253 TI - Hansen's disease: computed tomography findings in peripheral nerve lesions. AB - Hansen's disease induces hypertrophic neuropathy. Computed tomography was performed on ulnar, median, peroneal and tibial posterior nerves where there is clinically presumed lesion. The CT findings (diameter and density) have correlated well with intra-operative findings. The computed tomography is a reliable method to study peripheral nerve lesions in Hansen's disease patients. PMID- 2561254 TI - [The role of bitoxibacillin components in the manifestation of its biological activity]. AB - New methods were developed and applied to quantitative determination of beta exotoxin and antibiotic activity of delta-endotoxin with respect to Micrococcus spp. in bitoxibacillin (BTB) and the fermentation broths prepared under industrial conditions. The biosynthesis of beta-exotoxin in the period of its maximum accumulation during the fermentation was estimated. It was shown that the primary biological effect of BTB on insects consisted in the actions of beta exotoxin and delta-endotoxin. Biological activity of each of the entomocidal components of the entomocidal components of BTB did not practically correlated with the number of viable spores. There was a correlation between the antibiotic activity of crystalline B. thuringiensis subsp. thuringiensis solutions and the insecticidal activity of the entomopathogenic preparations. Determination of beta exotoxin and antibiotic activity of delta-endotoxin might be used as a complex procedure for testing the quality of BTB. The method for estimating antibiotic activity of the crystal solutions allowed one to assay the biological activity of other preparations based on Bacillus thuringiensis non-synthesizing beta exotoxin. PMID- 2561255 TI - [Evaluation of 30-year-activity of the Virology Laboratory of the Pasteur Institute in Madagascar (1958-1987)]. AB - The authors studied the different activities of Pasteur Institute virology laboratory since it was created. In general virology laboratory, 1.685 viral strains were isolated from 9.533 studied samples (17.6% of positivity). Enterovirus represented 90% of the viral strains. From 2.417 taking of rhino pharynx, 133 myxovirus were isolated: A/H3N2 represented 76% of them. 10.776 patients, from 36.169 antirabic department consultants, were vaccinated and 600 had an antirabic serotherapy. 2.828 suspect samples were analysed for the rabies diagnostic: 1.438 were positive (51%). The Institute produced 3.410 liters of Fermi human vaccine type. PMID- 2561256 TI - [Effect of a dehydrated extract of nopal (Opuntia ficus indica Mill.) on blood glucose]. AB - To assess if a dehydrated extract of nopal stems retains the effect on glycemia of the entire nopal stems two experiments were performed. A. Six patients with type II diabetes mellitus in fasting condition received 30 capsules containing 10.1 +/- 0.3 g of the extract, and serum glucose levels were measured hourly from 0 to 180 minutes. B. Six healthy volunteers received 30 capsules with the extract followed by 74 g of dextrose orally. Serum glucose measurements were made in a similar fashion. In each experiment a control test with empty capsules was performed. Nopal extract did not reduce fasting glycemia in diabetic subjects. Nevertheless, the extract diminished the increase of serum glucose which followed a dextrose load. Peak serum glucose was 20.3 +/- 18.2 mg/dl (X +/- SD) lower in the test with nopal than in the control one (P less than 0.025). Dehydrated extract of nopal (Opuntia ficus-indica Mill) did not show acute hypoglycemic effect, although could attenuate postprandial hyperglycemia. PMID- 2561257 TI - 4-Aminopyridine on the spontaneous contractions of isolated intestine. AB - The effects of 4-aminopyridine on isolated spontaneously active segments of rabbit jejunum were studied. The drug produced a dose-dependent increase of the tonic and phasic components of the contractile activity with an ED50 of 2 x 10( 5) M, lower than the reported dose for striated muscle. No effects were observed on the frequency of contractions at the concentrations of 4-aminopyridine used in this study. The results suggest both, an effect on the jejunal innervation and a direct action of the drug on the intestinal smooth muscle. The former mechanism includes the release of acetylcholine and is partially blocked by atropine. Direct effects of 4-aminopyridine on intestinal smooth muscle cells can be present at relatively low doses and they must be carefully evaluated during the suggested therapy of several human diseases with 4-aminopyridine. PMID- 2561258 TI - Preoperative embolisation in the management of juvenile nasopharyngeal angiofibroma. AB - Thirteen cases of juvenile angiofibroma were treated surgically with preoperative embolisation. One case occurred in a female. Embolisation significantly reduced blood loss and allowed complete surgical removal of the tumour in all cases. No significant complications occurred. Preoperative embolisation should be employed routinely in the surgical management of these patients. PMID- 2561259 TI - Albumin interferes with arachidonate metabolism in platelets and neutrophils. AB - The in vitro effects of albumin (150-600 microM) on arachidonate metabolism are compared in two types of circulating cells, platelets and neutrophils. The effect of this plasma protein on a functional aspect common to both these blood components, i.e. aggregation, was also investigated. Bovine serum albumin (BSA) significantly inhibited thromboxane B2 formation by washed platelets stimulated with threshold concentrations of collagen. In addition, this protein profoundly affected the formation of the major products via the lipoxygenase pathways, i.e., 12-hydroxyeicosatetraenoic acid and leukotriene B4 by platelets and neutrophils, respectively. This inhibitory effect was also confirmed for both types of cells considered on aggregation. Albumins of human origin and human plasma behaved similarly to BSA on the above parameters. Although the mechanism(s) responsible for the described effects require(s) additional studies, albumin, affecting platelet and neutrophil arachidonate metabolism, as well as the aggregatory response, might influence, in some conditions, cell activation. PMID- 2561260 TI - The transposable element Tn3 promotes general recombination at the neighboring regions. AB - Transposon Tn3 was inserted into a tRNA operon of the amber suppressor Su+2 on a transducing phage (lambda hcI857nin5pSu+2) by selecting phages with ampicillin resistance and Su- phenotypes. In a strain thus obtained, Tn3 was inserted between the promoter and the first tRNA gene of the operon, which was determined by DNA sequencing. The Su+2 tRNA operon on the transducing phage consisted of two tRNA genes for tRNA(Met) and Su+2 tRNA(2Gln), which was a deletion derivative of the supB-E tRNA operon of E. coli containing seven tRNA genes in the order of promoter-Met-Leu-Gln1-Gln1-Met-Gln2-Gln2. Proliferating the lambda hcI857nin5pSu+2::Tn3 in E. coli cells, a number of phages which had lost Tn3 were isolated, and their tRNA gene compositions as well as the DNA structures of the tRNA operon were analyzed. In many cases the tRNA genes which had been deleted from the original transducing phage were regained from the chromosomal supB-E operon. Thus the loss of Tn3 from the phages was not due to excision of the transposon but due to the replacement of a portion of the tRNA operon, including Tn3, with the host homologous region that did not contain Tn3. This type of replacement takes place rather efficiently as a consequence of Tn3 insertion, owing to the general recombination occurring between homologous tRNA genes of phage and host chromosomes in the presence of either host recA or phage red. No such enhanced recombination in a similar cross between phage and host chromosomes was observed with the Tn3 present in the trans position on an independent plasmid. We conclude that inserting Tn3 in cis promotes general recombination in the neighboring regions. Possible mechanisms for this new type of genetic effect of Tn3 are discussed. During the course of this study, a natural defective mutation (T11) was also detected in one of the duplicated tRNA(2Gln) genes in an E. coli K12 strain we used. PMID- 2561261 TI - Diversity of glucose entry routes in the Enterobacteriaceae. PMID- 2561262 TI - The ugp-encoded glycerophosphoryldiester phosphodiesterase, a transport-related enzyme of Escherichia coli. PMID- 2561263 TI - Carbon metabolism and catabolite repression in Rhizobium spp. PMID- 2561264 TI - [Iron as an inducer of nitric oxide formation in the animal body]. AB - Citrate iron complex injections to mice or rats resulted in the nitric oxide formation detected by nitric oxide binding to iron-diethyldithiocarbomate complexes. The mononitrosyl iron complexes formed were paramagnetic and EPR active. The maximal nitric oxide concentrations in rat livers were 15-20 nm per gram of tissue. Phenosan-K (an antioxidant) inhibited partly the iron capacity to nitric oxide formation in animal organisms. The nitric oxide formation was proposed to be due to some endogenic amino groups oxidation by active oxygen agents or products of lipid or non-saturated fatty acid production under the prooxidant action of the iron. PMID- 2561265 TI - [cAMP, calmodulin-dependent stimulation and stability to proteolysis of Ca 2+ transport in the heart sarcoplasmic reticulum]. AB - The calmodulin content in cardiomyocyte cytosol of hypoxic myocardium is increased compared to normal level. This is unaccompanied by differences in the stimulating effect of calmodulin on Ca2+ transport in sarcoplasmic reticulum (SR) of ischemic heart. The decrease of the endogenous cAMP-dependent protein kinase activity in ischemia is associated with the lowered resistance to trypsinolysis of Ca2+ transport in SR (trypsin/microsomal protein ratio is 1:10) with simultaneous Ca-ATPase activation. In the presence of exogenous protein kinase and cAMP the protective effect of phosphorylation on Ca2+ transport in SR vesicles of hypoxic cardiomyocytes treated with trypsin for 10 min reaches the same level as in intact heart. PMID- 2561266 TI - [Peptidylglycine alpha-amidating monooxygenase from bovine heart atrium secretory granules and adrenal chromaffin granules]. AB - About 40-60% of the peptidylglycine alpha-amidating amonooxygenase activity in the lysates of secretory granules from bovine atria and adrenal medulla isolated and lyzed in the presence of pepstatin, phenylmethylsulfonyl gluoride, N ethylmaleimide and catalase, was found to be in the soluble form. The remaining part bound to the membrane fraction was extracted with Triton X-100. The procedure of purification of the soluble form of peptidylglycine alpha-amidating monooxygenase from both atrial and chromaffin granules in electrophoretically homogeneous enzyme preparations was developed. The enzyme is made up of a single subunit with a molecular mass of 68 kDa and contains one copper atom per molecule. The EPR spectra of peptidylglycine alpha-amidating amonooxygenase and dopamine beta-monooxygenase were found to be practically identical, thus indicating that the copper environment in the both enzymes is the same. Both peptidylglycine alpha-amidating monooxygenase and dopamine beta-monooxygenase are inhibited by the neurocuprein apoform, an extremely acidic protein isolated from brain and secretory granules of different endocrine tissues. PMID- 2561267 TI - [Role of Cl-ionophore subunit in the functioning of GABA-benzodiazepine receptors]. AB - The displacement curves of the effect of picrotoxin on the S35-tert-butyl bicyclophosphorothionate binding to the brain membranes of inbred mice C57Bl/6 and Balb/c were analysed. The marked interstrain differences between modifications of IC50 and nHill dependent on the ionic force of the incubation medium were revealed. After partial stimulation of the receptor in the presence of 50 mM concentration of NaCl, the susceptibility of the receptor to picrotoxin was higher in the membrane suspension of C57Bl/6 mice than of Balb/c. Full stimulation of the receptor by 200 mM concentration of NaCl resulted in disappearance of interstrain differences. The value of nHill of C57Bl/6 membrane receptors increased after the change of the concentration of NaCl in the incubation medium from 50 to 200 mM, while it was invariable to Balb/c mice. It was marked that modification of the parameters of the binding of the radioligand to the membranes of C57Bl/6 mice was more latent, than for Balb/c mice. PMID- 2561268 TI - [C-src locus determines increased rate of Na+,K+-cotransport and increased calcium content in erythrocytes of (SHR x WKY) F2 hybrids]. AB - 26 male F2 hybrids between spontaneously hypertensive (SHR) and normotensive control (WKY) rats (SHRxWKY)F2 were segregated according to their c-src genotype into SS and WW homozygous groups, corresponding to SHR or WKY and WS heterozygous group. Na, K cotransport in erythrocytes in the WW group was equal to that of WKY and differs significantly from that of WS and SS groups (the rate of Na, K cotransport in latter groups was close to that of SHR). Ca content of RBC in WW group was equal to that of WKY, lower than that of WS and SS groups which in turn was significantly lower than in SHR, indicating polygenic control of the trait. Authors conclude that the c-src locus itself or some other loci inherited in conjunction with c-src determines both the increase of Na, K cotransport and calcium content in erythrocytes of SHR. PMID- 2561270 TI - [Cocaine and disturbances of adrenergic neurotransmission]. AB - Cocaine releases peripheral and central stores of catecholamines and their plasma concentration increases in very significantly under the influence of the drug. One also observes a vaso-constriction of cerebral and coronary arteries. Decreases in blood flow through those vessels have been observed in cocaine addicts examined with the magnetic resonance and positron emission tomography techniques. Some of these changes may still be present 15 days after withdrawal. The authors suggest an hypothesis to account for the general mode of action of cocaine. The synaptic increase in catechol produced by this drug is calcium dependent and would induce an "up regulation" of receptor proteins of calcium channels with increased calcium intracellular fluxes. There would be an increase in catecholamine neuronal turnover and synthesis not a depletion. Conversely the adrenergic post-synaptic receptors would undergo a "down regulation". Such impairment of adrenergic neurotransmission is associated with an increase in Angiotensin II release and a decreased GABA activity. Sensory stimuli resulting from this impairment induce a dominant memory associated with drug seeking and consuming behavior. PMID- 2561269 TI - Xylazine antinociception in mice: evidence for mediation by postsynaptic adrenoceptors. AB - The influence of 6-hydroxydopamine (6-OHDA) pretreatment on xylazine (XLZ) induced antinociception was studied in mice using the writhing test (60 mg/kg acetic acid, ip, as the algogenic compound administered 10 min after 0.5 and 0.75 mg/kg XLZ, sc). 6-OHDA (100 mg kg-1 injection-1 administered ip on days 1, 3, 5, 7, 9 and 11 after birth) did not modify XLZ-induced antinociception, suggesting that this effect is mediated by postsynaptic alpha-2 adrenoceptors. PMID- 2561271 TI - [Electrophoretypes of the genome of rotaviruses isolated from Senegalese children]. AB - Polyacrylamide gel electrophoresis allowing study of rotavirus genome was applied to 213 faecal specimens from children with acute gastroenteritis living in Dakar area, Senegal. We were able to define an electrophoretype in 41 cases (19.2%). Five different RNA electrophoretypes were shown. The electrophoretype 4 was the most frequent (41.4%). "Long" and "short" electrophoretic migration patterns were found in 36 and 5 cases, respectively. We found 34.1% of electrophoretypes in children aged from 6 to 11 months. The electrophoretypes 1, 3, 4 and 5 have co circulated throughout the study period (March 1985 to August 1986). No mixed infections were found. Large surveys of this type would allow to evaluate the general situation in Senegal in urban areas as well as in rural areas. PMID- 2561272 TI - [Chronic diarrhea and parasitoses in adults suspected of AIDS in the Ivory Coast]. AB - 148 adult patients with chronic diarrhoea and suspected to be HIV infected have had stool examinations. 46 are without any enteric parasite. Those detected in the others patients are Sporozoans: I. belli (16.2%) and Cryptosporidium sp. (6.7%) found alone or joint, together or with other parasites. Among those, all Flagellate species are identified, but T. intestinalis (6%) is predominant. Entamoeba coli (8.%) is the most frequent amebic species, however, E. histolytica histolytica have been found twice, once singly, the second associated with I. belli, Schistosoma mansoni and Candida albicans. Necator americanus (14%) and Strongyloides stercoralis (12%) are the predominant worm species. Among the yeasts, C. albicans (35.8%) is the most important species isolated, singly in 13.5% of the patients. In an intertropical and parasitical endemic area where many parasites are not considered uncommon, opportunist agents as I. belli, Cryptosporidium sp. and C. albicans appear in an non-negligible frequency in our study. PMID- 2561273 TI - Diethyldithiocarbamate (DEDC) enhances quinone mediated oxidative stress cytotoxicity in isolated hepatocytes by forming toxic quinone conjugates. AB - The copper-chelating thiol drug, diethyldithiocarbamate (DEDC) had previously been used to inhibit superoxide dismutase (SOD) and enhance oxidative stress mediated cytotoxicity. Using isolated rat hepatocytes, it was confirmed that DEDC enhances oxidative stress cytotoxicity induced by 1,4-naphthoquinone (1,4-NQ) and 1,4-naphthoquinone-2-sulphonate (1,4-NQ-2S). However, equimolar concentrations of DEDC also enhances cytotoxicity induced by benzoquinone, previously shown to cause cytotoxicity as a result of alkylation and not oxidative stress. Higher DEDC concentrations on the other hand protected against benzoquinone-induced cytotoxicity. Finally, the susceptibility of hepatocytes to quinone mediated oxidative stress cytotoxicity was not enhanced if the DEDC was removed before incubating the hepatocytes with naphthoquinone or benzoquinone. Enhanced oxidative stress cytotoxicity was only observed if the DEDC was present when hepatocytes were treated with quinones. It was concluded that DEDC forms conjugates with quinones which undergo futile redox cycling in the hepatocyte and form H2O2 as well as increase the susceptibility of hepatocytes to H2O2. PMID- 2561274 TI - Effect of sodium valproate on the secretion of proopiomelanocortin derived peptides from cultured rat anterior pituitary cells. AB - We examined effects of sodium valproate, a gamma amino butyric acid (GABA) transaminase inhibitor, on the secretion of immunoreactive (IR)-ACTH and IR-beta endorphin/LPH from cultured rat anterior pituitary cells to determine whether sodium valproate has a direct action on the secretion of ACTH and its related peptides from the cultured rat anterior pituitary gland. During the 3 h incubation, the basal secretion of IR-ACTH and IR-beta-endorphin/LPH decreased to 50.8% and 58.3%, respectively, of the control concentration after adding 10(-7) M sodium valproate into the incubation media and to 67.7% and 69.3%, respectively, of the control levels with 10(-8) M sodium valproate. However, sodium valproate at a concentration of 10(-6) M or 10(-9) M did not affect the basal concentration of IR-ACTH and IR-beta-endorphin/LPH. Sodium valproate at a concentration of 10( 7) M significantly attenuated the stimulated release of IR-ACTH and IR-beta endorphin/LPH by 10(-9) or 10(-10) M of ovine corticotrophin releasing factor. These results indicate that sodium valproate could directly effect rat anterior pituitary cells to suppress both basal and stimulated release of proopiomelanocortin derived peptides and this supports the hypothesis that sodium valproate has a direct effect at the pituitary corticotroph in reducing plasma ACTH. PMID- 2561275 TI - Diabetic state-induced activation of calcium-activated neutral proteinase in mouse skeletal muscle. AB - The effect of a diabetic state in the diabetic KK-CAy mouse on calcium activated neutral proteinase (CANP) of hind-limb skeletal muscles was investigated. In the diabetic state, there was an increased sensitivity to activation of CANP by calcium (Ca). In addition, there was an enhancement of maximal activity of the enzyme. The effect was induced by secondary modification of the diabetic state, but not genetical factors. Several lines of evidence suggest that the CANP is responsible for 92 K dalton protein in diabetic skeletal muscles. Among the evidence are the following: a) The 92 K band in the diabetic muscles was lower than in the prediabetic mouse and restored by the addition of 2 mM EDTA and 2 mM EGTA. b) The band was reduced by increasing the Ca content and neutral pH in the non-diabetic normal muscles. c) E-64-C, a CANP inhibitor, restored the 92 K component reduced by the diabetic state. Since the band in denervated muscles was not changed by the Ca chelating agents, the reduction of the band in the diabetic muscles is related with musculotrophic factors, not diabetic neuropathy. These results suggest that diabetic amyotrophy may be regarded as a phenomenon linked to an increase in intracellular Ca ions and an increase in CANP activity. PMID- 2561276 TI - The possible role of endogenous digitalis-like substance in the regulation of circadian changes in urinary electrolyte excretion in man. AB - The urinary volume (U.V.), Na excretion (UNaV) and K excretion (UKV) have been reported to show a circadian rhythm in man, but the mechanism of this rhythm has not been made clear. To investigate how atrial natriuretic peptide (ANP) and endogenous digitalis-like substance (DLS) participate in the circadian change in urinary electrolyte, the circadian changes in ANP and DLS (digoxin-like immunoactivity: DLI, Na-K-ATPase inhibitor: ATPI, ouabain binding inhibitor to Na K-ATPase: OBI) were evaluated in 5 normal man. ANP, DLI and OBI showed no significant correlation with urinary electrolyte excretion, but there was a significant positive correlation between plasma ATPI and urinary Na excretion. From these results it is suggested that circulating Na-K-ATPase inhibitor (plasma ATPI) may be involved in the regulation of the circadian rhythm of urinary Na excretion. PMID- 2561277 TI - Evidence for age-related change in plasma 19-hydroxyandrostenedione. AB - The steroid, 19-hydroxyandrost-4-ene-3, 17-dione (19-hydroxyandrostene-dione, 19 OH-A-dione) has been known to enhance the mineralocorticoid action of aldosterone. To investigate the age-related change in the plasma 19-OH-A-dione concentration, plasma 19-OH-A-dione, androst-4-ene-3, 17-dione (A-dione), aldosterone and cortisol of 38 non-hypertensive healthy subjects (18 young men and 20 aged men) measured by specific radioimmunoassays. The basal plasma 19-OH-A dione and A-dione concentration in aged men was significantly lower than in young men (P less than 0.01). Moreover, there was found to be a positive correlation between plasma 19-OH-A-dione and A-dione (P less than 0.01). On the other hand, plasma aldosterone and cortisol in aged men showed a tendency to decrease, but no statistical significance compared to young men was observed. This study demonstrated that there was an apparent age-related decrease not only in plasma A dione, but also in plasma 19-OH-A-dione, an amplifier or aldosterone action. PMID- 2561278 TI - Structural and functional stability of foreign genes in transgenic tobacco plants. AB - The Drosophila genomic fragment Dm111 and the selectable dominant nptII gene were transferred via a Ti-vector into tobacco plants in order to check the structural and functional stability of transgenes in plants and their progeny. Southern blot analyses clearly showed that transgenes were integrated intact and did not suffer from any gross DNA rearrangements. Contrary to this structural stability, not all of the transgenic plants and their offspring displayed the original and stable expression of the nptII gene. The levels of the NPTII enzyme strongly varied in individual plants and did not depend on the copy number of the nptII gene. Both the non-stable nptII expression during the individual plant development in one original transgenic tobacco and some irregularities in segregation ratios after self-pollination indicated that epigenetic effects due to methylation of DNA modulated the expression of foreign genes in transgenic plants. This conclusion was supported by a spontaneous and 5-azacytidine-stimulated demethylation. PMID- 2561279 TI - Immunoglobulin heavy chain genes in the hybridoma PTF-02. PMID- 2561280 TI - [Modern knowledge of the biology of cancer]. PMID- 2561281 TI - [The endogenous opioid system and selective attention]. AB - The effects of fentanyl and naloxone upon the late components (P150 and P300) of auditory evoked potentials were examined with the purpose of establishing its electrophysiological mechanism of action. 50 adult patients were evaluated by means of three pharmacological paradigms: 1. Initial control (C), saline solution (S) and final control (C'). 2. C, fentanyl (F) and C'. 3. C, naloxone (N) and C'. Fentanyl decreased the amplitude of components P150 and P300, as well as patients' performance; while naloxone increased the amplitude of P150 and small P300 (S), having opposite effect upon the large P300 (L). It is concluded that fentanyl caused a global disminution of patients' attention, while naloxone improves attention in patients with P300S and depresses subjects with P300L. PMID- 2561282 TI - Effect of exogenous 5-hydroxytryptamine on pathogenicity of Entamoeba histolytica in experimental animals. AB - In an effort to find out the mechanism(s) operative in enhancing the pathogenicity of E. histolytica in hosts under heat stress reported earlier, effect of 5-hydroxytryptamine (5-HT) on the virulence of the parasite was examined in just weaned Charles Foster strain of albino rats. Pathogenicity of 10 strains of E. histolytica, from various forms of intestinal amoebiasis, grown in modified Boeck and Drbohlav's medium was assessed by caecal scoring. Administration of 5-HT in infected animals significantly enhanced the pathogenicity of all the seven strains tested. Treatment of the host with the 5 HT precursor L-tryptophan also increased the caecal scores examined with three strains of E. histolytica. Prior blocking of tissue 5-HT receptors by administration of methysergide almost completely abolished the pathogenicity enhancing effect of 5-HT treatment. This suggested that 5-HT itself and not any of its metabolites was responsible for the observed increase in pathogenicity of E. histolytica on 5-HT treatment of the host. PMID- 2561283 TI - Technetium-99m pyrophosphate myocardial scintigraphy in the diagnosis of acute rheumatic carditis. PMID- 2561284 TI - Rotavirus vaccines. PMID- 2561285 TI - Influence of eicosapentaenoic acid and vitamin E on hepatic hydroxyproline content in rabbits fed cholesterol-rich diet. AB - The effect of eicosapentaenoic acid (EPA) and vitamin E on hepatic hydroxyproline content, as an index of collagen was examined in rabbits receiving cholesterol rich diets for a period of 45 days. Rabbits were divided as control (A) and cholesterol fed groups (B, C, D). Group C received 80 mg. of EPA and group D received 100 IU of vitamin E daily in addition to the cholesterol rich diet (2% w/w) which was solely given to group B. The maintenance of rabbits on high cholesterol diets resulted in significantly increased liver cholesterol concentrations. This effect was most pronounced in rabbits receiving cholesterol alone. Hepatic triglyceride levels remained unchanged in all cholesterol-fed rabbits, but total phospholipid levels in liver significantly decreased in EPA and vitamin E supplemented rabbits. An interesting finding was the increase in hepatic hydroxyproline content in rabbits following the administration of EPA and vitamin E to cholesterol rich diet. PMID- 2561286 TI - Liver cholesterol concentrations in mice fed diets containing various sources of fat, carbohydrates or fiber. AB - Liver cholesterol concentrations were measured in mice after feeding for 30 days cholesterol-free, semipurified diets containing various sources of fat, carbohydrates or fiber. Olive oil produced significantly higher liver cholesterol concentrations than tallow, sunflowerseed oil and cocoa fat. In mice fed either fructose or sucrose liver cholesterol was significantly increased when compared with mice fed galactose or lactose. Dietary cellulose, when compared with pectin, did not influence liver cholesterol. The amount of fat in the diet, in the form of either corn oil or coconut fat, had no significant effect on liver cholesterol. It is concluded that the type of carbohydrate and fat in the diet are major determinants of liver cholesterol in mice. PMID- 2561287 TI - The nature and function of polypeptide growth factor receptors in the human placenta. PMID- 2561288 TI - Expression of the gene encoding cytochrome c3 from Desulfovibrio vulgaris (Hildenborough) in Escherichia coli: export and processing of the apoprotein. AB - The expression of cytochrome c3 from Desulfovibrio vulgaris (Hildenborough) was examined in Escherichia coli transformed with either of two plasmids, pJ8 and pJ81. The former has an 840 bp insert of D. vulgaris DNA, containing the structural gene for cytochrome c3 (387 bp) and its promoter region. Plasmid pJ81 was generated from pJ8 by deoxyoligonucleotide-directed mutagenesis to direct the synthesis of a protein with an altered signal peptidase cleavage site [Ala(-1)--- Asp(-1)]. Synthesis of the 14 kDa precursor, which was partly processed to the 12 kDa mature protein, was observed in cells of E. coli TG2(pJ8) by SDS gel electrophoresis and Western blotting. Analysis of spheroplasts revealed that the processed polypeptide was present in the periplasm while the precursor was found only in the membrane/cytoplasmic fraction. No processing was observed in E. coli TG2(pJ81) cells, due to the mutation of the signal peptide cleavage site. No insertion of haem into the E. coli product could be detected in E. coli TG2(pJ8) cells by post-electrophoretic protohaem fluorescence analysis. The sensitivity of the cytochrome c3 synthesized in E. coli TG2(pJ8) to digestion by chymotrypsin also indicated that the apoprotein was formed. The results indicate that E. coli is capable of synthesizing and exporting the cytochrome c3 polypeptide, but fails to insert the haems. PMID- 2561289 TI - Transposon mutagenesis and complementation of the fructokinase gene in Rhizobium leguminosarum biovar trifolii. AB - Transposon Tn5 was used to generate a fructokinase mutation in Rhizobium leguminosarum biovar trifolii BAL. The section of the genome containing Tn5 was cloned into the EcoRI site of the vector pHC79 and isolated by direct selection on medium containing kanamycin and tetracycline. Total EcoRI digestion was used to obtain a single fragment containing Tn5 and flanking DNA sequences. The flanking DNA was used as a probe to isolate an intact fructokinase gene from a pLAFR1 cosmid clone bank of the parental strain. A cosmid showing homology to the probe was tri-parentally conjugated into the fructokinase-negative strain, complementing the mutation. The complemented mutant exhibited the wild-type phenotype, with an increase in fructokinase production presumably due to multiple copies of the gene. PMID- 2561290 TI - Induction of phosphatidylcholine biosynthesis via CDPcholine pathway in lung and liver of rats following intratracheal administration of DDT and endosulfan. AB - The induction of phosphatidylcholine (PC) biosynthesis via the CDPcholine pathway in lung and liver of rats has been shown following the intratracheal administration of 1,1,1-trichloro-2m2-bis(p-chlorophenyl) ethane (DDT) (5 mg/100 g body weight) and endosulfan (1 mg/100 g body weight) for 3 days. Controls received only the vehicle solution (groundnut oil, 0.1 m1/100 g body weight). The treatment of DDT and endosulfan significantly increased the PC contents and the incorporation of radioactive [methyl-3H]choline into PC of lung and liver microsomes. The incorporation of radioactive [methyl-14C]methionine into microsomal PC of lung and liver was not affected significantly by treatment with either of the insecticides. 1,4,5,6,7-hexachloro-5-norbornene-2,3-dimethano cyclic sulfite (endosulfan) administration significantly increased the activity of choline kinase and phosphocholine cytidylyltransferase (both cytosolic and microsomal) of lung, whereas DDT increased the activity of only latter. In liver, both DDT and endosulfan administration significantly increased the activity of choline kinase and phosphocholine cytidylyltransferase (both cytosolic and microsomal). However, the activity of phosphocholinetransferase was not affected in both lung and liver microsomes of rats treated with these insecticides. The PC precursor pool sizes, choline and phosphorylcholine, of lung and liver tissues were not altered by DDT and endosulfan treatments. The present results suggest that the increased level of PC and incorporation of radioactive [methyl 3H]choline into microsomal PC could be the result of increased activity of choline kinase and phosphocholine cytidylyltransferase of lung and liver of rats following intratracheal administration of DDT and endosulfan. PMID- 2561291 TI - Immunotherapy and enhanced antibody-dependent cell-mediated cytotoxicity using virally-infected target cells. AB - We examined the ability of in vitro addition of Interleukin-2 (IL-2) to differentially enhance antibody-dependent cell mediated cytotoxicity (ADCC) utilizing cultured Epstein-Barr virus infected cells and gammaglobulin (Sandoglobulin). We found significant enhancement of ADCC when IL-2 was added. Chronic Epstein-Barr virus or Chronic Fatigue Syndrome patients in a therapeutic gammaglobulin program may benefit from IL-2 given in vivo. PMID- 2561292 TI - Subject-reported compliance in a chemoprevention trial for familial adenomatous polyposis. AB - A high level of compliance with an assigned treatment regimen is fundamental to accurate assessment of treatment effectiveness in any clinical trial. If compliance is poor, an effective treatment may be confounded by inadequate delivery of the regimen. Although much research has focused on broad aspects of compliance dealing with clinical therapeutic situations, there was a need for further research dealing specifically with adherence issues in a long-term chemoprevention trial since subject motivation in the latter is likely to differ from that of the former. Examining subject-reported compliance over the first 2 year treatment periods of a long-term chemoprevention trial for familial adenomatous polyposis, it was found that (1) compliance decreased over time, (2) fiber compliance was lower than vitamin compliance, and (3) four explanatory variables which may be amenable to individualized study-team interventions emerged as useful prognosticators of fiber compliance. PMID- 2561293 TI - Studies on several naturally occurring lignans. AB - Form the bark of Taiwania cryptomerioides, the flower buds of Magnolia fargesii, the wood of Juniperus formosana, and the nutmeg of Myristica fragrans and Myristica cagayanensis, eleven new kinds of lignans and neoli gnans were isolated. The structures of these compounds were elucidated by physical and chemical methods. PMID- 2561294 TI - [Glomus tumor--clinical experience of 14 cases]. AB - Fourteen cases of subungual glomus tumors were treated from June 1, 1984 to November 15, 1987. Of these cases, 4 were male and 10 were female; 8 occurred on the; right hand and 6 on the left hand; tumors found on the thumb: 6 case; on the index: 1 case; on the middle finger: 1 case, and on the ring finger: 6 cases. All these tumors were subungual, located around the lunula area and were treated by surgical excision with complete relief of symptoms. In the earlier cases, the tumors were approached with total nail removal and longitudinal incision in the nail bed. After the tumors were excised, the nail beds were repaired with 5 "0" or 6 "0" absorbable suture material. Longitudinal ridge deformities of the nail were noted in all these cases in the follow-up. In the later cases, the tumors were approached with proximal half nail removal and a "U" or "+" incision in the nail bed. After the tumors were excised, the nail beds were repaired with 7 "0" or 8 "0" suture material. No deformities of the nail were noted in these cases in the follow up. We suggest the subungual tumor be treated by surgical excision, and to avoid later nail deformity, the tumor located around the lunula be approached by proximal half nail removal, a "U" or "+" incision on the nail bed and repair of the nail bed with 7 "0" or 8 "0" suture material. PMID- 2561295 TI - Chronic myelomonocytic leukemia with polyneuropathy and IgA-paraprotein. AB - A 65-year-old woman had chronic myelomonocytic leukemia with peripheral neuropathy and IgA paraprotein with kappa type light chain. Plasma cells with "flaming" cytoplasma were seen in the bone marrow specimens. The findings suggest that chronic myelomonocytic leukemia may involve B-lymphocytes in the proliferative process and that it may be a clonal disease. Moreover, this case was associated with peripheral sensori-motor polyneuropathy. The disorder was responsive to prednisolone, as oral administration of prednisolone improved not only the peripheral polyneuropathy, but also the hematological findings. Serum vitamin B12 and lysozyme in the serum and urine were decreased by the steroid therapy. The administration of prednisolone is effective not only for polyneuropathy but also chronic myelomonocytic leukemia. PMID- 2561296 TI - The discrepancy between chemotaxis and leukotriene B4 production in a patient with chronic neutrophilic leukemia. AB - Chronic neutrophilic leukemia (CNL) is a rare type of leukemia. We diagnosed a 81 year-old woman as CNL because she showed that sustained leukocytosis dominated by mature neutrophils, hepatosplenomegaly, high neutrophilic alkaline phosphatase (NAP) score, absence of the Ph1 chromosome and no evidence of leukemoid reaction. During the clinical course, she did not manifest hemorrhagic tendency or infection. We also examined neutrophilic function including chemotaxis, chemiluminescence, nitroblue tetrazolium (NBT) dye reduction, which all indicated normal neutrophil function. Using a reversed phase-high pressure liquid chromatography (HPLC), we detected the production of leukotriene B4 (LTB4) in neutrophils. We found that the LTB4 production was decreased in neutrophils whereas they showed normal chemotaxis. This discrepancy has never, to our knowledge, been reported before in case of CNL. PMID- 2561297 TI - Peripheral nerve involvement in HTLV-I-associated myelopathy: report of a case. AB - A patient came to our clinic suffering from mild sensory loss, exaggeration in knee reflexes, bilateral extensor plantar responses and elevation of the antibody level to the human T-lymphotropic virus type I (HTLV-I). Electrophysiological and pathological studies showed axonal and demyelinating neuropathy. The condition appeared to be a slowly progressive myeloneuropathy. HTLV-I may play an important role in the pathogenesis of myeloneuropathy. PMID- 2561298 TI - Adrenocorticotropic hormone (ACTH). PMID- 2561299 TI - Diagnosis of human papillomavirus infection in uterine cervix by immunoperoxidase technique. AB - The present study detected human papillomavirus (HPV) genus-specific antigen in paraffin section of chronic cervicitis tissue by peroxidase-antiperoxidase (PAP) technique. The results showed that HPV antigen was detected in 34 of 101 specimens (33.66%). All the normal control sections were negative. The antigen positive nuclei were found in the koilocytes of the upper layers of the epithelium. Antigen positive cells may display a uniform or a scattered pattern. The pathologic feature of HPV infection discribed by Meisels et al. was found in all positive sections. It is suggested from the results that almost one third of chronic cervicitis is induced by HPV infection. PMID- 2561300 TI - [Expression of lactoferrin and myeloperoxidase genes in blasts of acute lymphoblastic leukemia]. AB - The myeloperoxidase (MPO) and lactoferrin genes are strictly associated to the granulocytic differentiation pathway. Many reports show that immunologic and molecular cell markers, though to be lineage specific, are in fact expressed in cells of different lineages. We have identified lymphoid blast cell populations expressing the MPO gene at the mRNA level, but lacking any detectable MPO protein. We addressed this issue studying the expression of the MPO and lactoferrin genes in extremely homogeneous Acute Lymphoblastic Leukemia (ALL) blast cell populations. The study was carried out by solution hybridization of total cellular mRNA extracted from the leukemic populations with an antisense RNA transcribed by riboprobes. Either MPO or lactoferrin mRNAs were detectable in some ALL populations. PMID- 2561301 TI - [Primary carcinoma of the liver and hepatitis C virus in Italy. A prospective study in patients with cirrhosis]. AB - The possibility to detect the antibody to hepatitis C virus (HCV) has allowed to estimate the prevalence of this virus in patients with hepatic disease, mostly in those with hepatitis considered non-A non-B. Literature shows that HCV causes about 75% of cases of cryptogenic hepatitis and more than the 90% of post transfusional hepatitis. Circumstantial evidence suggests the existence of a relationship between parenterally-transmitted non-A non-B hepatitis (PTH) and primary liver cancer (PLC). With the advent of anti-HCV, it is now possible to assess directly whether or not there is a relationship between PTH and PLC. So anti-HCV was looked for in the sera of 365 patients with cirrhosis prospectively followed-up for early detection the development of PLC, using an enzymatic immunoassay (ELISA Ortho DS). At baseline anti-HCV was detected in 221 patients (60%). During 5-39 month 53 patients developed PLC and anti-HCV was detected in 68% of them. The univariate analysis demonstrated that alcohol abuse, anti-HBs and anti-HBc were the only covariates that were significantly associated with an increase risk of developing PLC. When these factors were introduced in the step wise regression analysis, age and alcohol were found to be the only independent risk factors. The high prevalence of anti-HCV found in patients with cirrhosis and PLC suggests that HCV might play a role in this tumor; the frequent co occurrence of HCV and HBV markers suggests that HCV-HBV coinfection might be pathogenically important; alcohol was the most important non-viral risk factor for PLC. PMID- 2561302 TI - Inhibition by pertussis toxin of guanyl nucleotides exchange on transducin in bovine rod cell membranes. AB - The effect of pertussis toxin on GTP-binding protein of bovine rod cell outer segments (transducin) was studied. Pertussis toxin was shown to ADP ribosylate either alpha subunit of free transducin or transducin-GDP complex, whereas GTP and its analogue Gpp(NH)p strongly inhibit ADP ribosylation of transducin. Pertussis toxin inhibits rod outer segment membrane GTPase and GTPase of homogeneous transducin by 40% and 70-80%, respectively. Activation of rod cell cyclic nucleotide phosphodiesterase by transducin is reduced after its preincubation with pertussis toxin. In transducin modified by pertussis toxin, 83% of GDP becomes tightly bound and cannot be exchanged with Gpp(NH)p. The stabilization of complex transducin-GDP after ADP ribosylation can explain the inhibitory effect of pertussis toxin on GTP hydrolysis by transducin, and on phosphodiesterase activation by guanyl nucleotides. PMID- 2561303 TI - Protein kinases in neutrophils: a review. AB - In chemotactic factor-stimulated neutrophils, rapid increases of intracellular levels of cyclic AMP, calcium, and diacylglycerol have been observed and may be linked to protein kinase activation. The study of the physiological role and regulation of protein kinases in the neutrophil and the identification of their substrates has provided valuable information on the molecular mechanism of neutrophil activation. The focus of this review is on those aspects of protein kinases that are relevant to neutrophil activation and on the substrate proteins for these protein kinases. The possible role of protein phosphorylation in neutrophil function is also discussed. PMID- 2561304 TI - Changes in differently processed soya bean (Glycine max.) and lima bean (Phaseolus lunatus) with particular reference to their chemical composition and their mineral and some inherent anti-nutritional constituents. AB - The effects of 3 processing techniques: cooking, roasting and autoclaving on the proximate chemical composition, the mineral content and some inherent toxic factors of soya and lima beans were investigated. The processing techniques generally reduced the crude fibre levels and enhanced the extractable fat in the soya and lima beans. The coefficients of variability for crude fibre and ether extract due to the processing techniques of soya and lima beans were 20.9%, 16.0% and 22.3%, 38.1%, respectively. In parallel with decreased ash content in the cooked bean samples, there was a decrease in the K, Mg, Na and P levels relative to the raw bean and also relative to the other processing techniques. Mineral contents of the autoclaved bean samples were generally similar to those of the raw (unprocessed) samples. Under the processing conditions, roasting caused the highest reduction in thioglucoside content (59%) in soya bean while cooking caused the highest reduction in lima bean (78%). Trypsin inhibitor activity (TIA) ranged between 0.59 mg/g sample in the cooked soya bean and 11.6 mg/g sample in the raw bean while the corresponding values for lima bean ranged between 0.59 and 6.3 mg/g sample. Cooking and roasting caused over 90% reduction of TIA, while autoclaving caused 64-69% reduction in both bean samples. Under the assay conditions, haemagglutinating activity was not detected in the cooked and autoclaved soya and lima beans. The need to prevent both functional and nutritional damage to food proteins and other nutrients, resulting from excessive heating, was discussed. PMID- 2561305 TI - Compositional evaluation of some cowpea varieties and some under-utilized edible legumes in Nigeria. AB - The nutritive potentials of some cowpea varieties such as Ife Brown, Ife Bimpe, IT84E-124, K59 and TVX716 and some under-utilized edible legumes grown in Nigeria such as pigeon pea (Cajanus cajan), lima bean (Phaseolus lunatus), lablab bean (Dolichos lablab), mucuna bean (Mucuna sp.) and Sphenostilis sternocarpa have been evaluated with respect to their proximate chemical composition, mineral content and some endogenous toxic constituents. The cowpea varieties contained on the average 22.5 g crude protein (CP), 2.60 g crude fibre (CF), 5.89 g either extract (EE) and 3.36 g ash/100 g DM while the under-utilized legumes contained 21.7, 6.10, 2.86, and 3.56 g/100 g DM for CP, CF, EE and ash respectively. Distinct varietal differences were observed for EE values as indicated by the coefficients of variation (CV) of 102% for cowpea and 60.8% for the under utilized legumes. The CF content of the under-utilized legumes were generally higher than those of the cowpea varieties. Potassium was the most abundant mineral in both the cowpea varieties and the under-utilized legumes with mean values of 1.45 and 1.66% respectively, while P was the least abundant with 13.1 and 8.50 ppm, respectively. There were marked intra-varietal differences in the P content as shown by the high CV of 84.0 and 73.9% for the cowpea varieties and the other legumes. The cowpea varieties generally had higher levels of thioglucosides, trypsin inhibitor activity (TIA) and lower haemagglutinating activity (mean values of thioglucosides: 3.86%, of TIA: 13.9 mg/g protein and of haemagglutinating activity: 13.0 HU/mg N respectively), than the under-utilized legumes with mean respective values of 1.22%, 9.84 mg/g protein and 22.7 HU/mg N. The nutritional implications of these anti-nutritional components were discussed and some reasons adduced for the under-utilization of some of these legumes inspite of their apparent similarity in nutritional quality to the more commonly consumed grain legumes. PMID- 2561306 TI - Activation of protein kinase C via the T-cell receptor complex potentiates cyclic AMP responses in T-cells. AB - We have recently shown that activation of protein kinase C by tumour promoting phorbolesters, such as 4 beta-phorbol-12,13-dibutyrate, stimulates adenosine induced accumulation of cAMP in Jurkat cells, a human T-leukaemia line. Activating the CD3 complex associated with the T-cell receptor by means of the monoclonal antibody OKT3 caused a concentration-dependent accumulation of inositol phosphates and an increase in the phosphorylation of an endogenous protein kinase C substrate. OKT3 also mimicked the previously reported effects of protein kinase C since it potentiated the cAMP stimulation by either an adenosine analogue, NECA, or cholera toxin. Thus, our results indicate that stimulation of a receptor activating phospholipase C and protein kinase C can secondarily enhance the action of agonists that act on adenylate cyclase-coupled receptors. PMID- 2561308 TI - Effects of suramin and alpha, beta-methylene ATP indicate noradrenaline-ATP co transmission in the response of the mouse vas deferens to single and low frequency pulses. AB - Vasa deferentia from mice were field-stimulated by trains of 10 pulses delivered at 0.5 Hz. The pulses elicited separate twitches, the first of which (corresponding to a single pulse) exceeded the following ones in height and width and often was clearly biphasic. alpha, beta-Methylene-ATP 1 mumol/l and suramin 100 mumol/l caused almost identical changes. They reduced the height of the first twitch in the train by about one half and also reduced its width in a manner indicating that only the second, slow phase remained, but reduced much more markedly the following twitches in which now a small second, slow phase also became detectable. Idazoxan 0.1 mumol/l or yohimbine 0.1 mumol/l, when added in the presence of alpha, beta-methylene-ATP or suramin, further decreased the first twitch but enhanced twitches No. 2 to 10. These responses were then almost abolished by prazosin 0.1 mumol/l. Successive addition of prazosin 0.1 mumol/l and idazoxan 0.1 mumol/l to previously untreated vasa deferentia depressed the response to the first pulse by about one half in a manner indicating that only the first, rapid phase remained, but had comparatively little effect on the responses to the subsequent pulses. Suramin 100 mumol/l almost abolished the contractions remaining in the presence of prazosin and idazoxan. The results indicate that the first, rapid phase of the neurogenic contractions elicited by single or low frequency pulses is mediated by ATP which substantially contributes to all responses in a train. The second, slow phase is mediated by noradrenaline which substantially contributes to the response to the first pulse only.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561307 TI - [3H]HOE166 defines a novel calcium antagonist drug receptor--distinct from the 1,4 dihydropyridine binding domain. AB - Benzothiazinones represent a novel class of drugs which block voltage-dependent L type calcium channels in different tissues. [3H]HOE166 (R-(+-)-3,4-dihydro-2 isopropyl-4-methyl-2-[2-[4-[4-[2- (3,4,5-trimethoxyphenyl)ethyl] piperazinyl]butoxy]phenyl]-2H-1,4- benzothiazin-3-on-dihydrochloride; approximately 57 Ci/mmol) a potent optically pure benzothiazinone was employed to characterize receptors associated with skeletal muscle transverse tubule calcium channels. [3H]HOE166 reversibly labels the membrane-bound calcium channels with high affinity (Kd = 0.36 +/- 0.05 nM; Bmax = 18.2 +/- 3.3 pmol/mg of membrane protein; means +/- SD, n = 13), HOE166 (Ki = 0.76 nM) is 29-fold more potent than the respective (S)-enantiomer (Ki = 22.1 nM). Binding is inhibited by divalent and trivalent cations (Cd2+ and La3+ being most potent) and other calcium channel drugs (1,4 dihydropyridines, phenylalkylamines, benzothiazepines). High affinity [3H]HOE166 binding activity is maintained (Kd = 4.5-9.0 nM) after solubilization and purification (554-1350 pmoles/mg of protein) of the calcium channel complex from transverse-tubule membranes. The following data support our recent claim (Striessnig et al. 1985, 1988) that HOE166 labels a domain on L-type calcium channels which is distinct from that defined by 1,4 dihydropyridines, phenylalkylamines or benzothiazepines: (1) All 1,4 dihydropyridine-, phenylalkylamine- and benzothiazepine-receptor-selective drugs tested are only very weak inhibitors of [3H]HOE166 binding. (2) (+)-PN200-110 only partially inhibits [3H]HOE166 binding to the purified calcium channel complex.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561309 TI - Eicosanoids and peripheral neurotransmission. PMID- 2561311 TI - [Brazilian Cooperative Group for the Treatment of Wilms' Tumor]. PMID- 2561310 TI - The effects of plasma lipoproteins on platelet responsiveness and on platelet and vascular prostanoid synthesis. PMID- 2561312 TI - [The role of adenosine cyclic monophosphate (c-AMP) in the pathogenesis of epilepsy and the mechanism of the action of anti-epileptic drugs]. AB - Modern views on the role of c-AMP and c-CMP in the regulation of metabolism and function of the neuron and synaptic transmission are presented. A hypothesis is put forward of the participation of these cyclic nucleotides in the control of epileptogenic focus through modulation of the resting potential of the neuron. PMID- 2561313 TI - [Malignant mixed tumor of the breast]. PMID- 2561314 TI - [Medical treatment of hyperthyroidism]. PMID- 2561315 TI - Lipoyl-containing components of the pyruvate dehydrogenase complex: roles in modulating and anchoring the PDH kinase and the PDH phosphatase. PMID- 2561316 TI - Bicarbonate, chloride, and proton transport systems. PMID- 2561317 TI - Pumps and pathways for gastric HCl secretion. AB - Data reviewed herein show that the HCl-secreting parietal cell is an exaggerated example of dynamic membrane transformation. Recruitment and recycling of membrane provide the means for the massive redistribution of the gastric proton pump, the H,K-ATPase, from one membrane domain (cytoplasmic tubulovesicles) to another (apical plasma membrane) as a function of parietal cell activation and inactivation. Functional activation of HCl secretion requires not only the redistribution of pump protein, but also the participation of pathways for the rapid flux of K+ and Cl- across the apical membrane. In apical plasma membrane vesicles from stimulated cells these pathways appear to be conductive and can operate independently. Thus, our model for the parietal cell proposes that K+ and Cl- flux from cell to lumen, operating in parallel and in concert with ATP-driven H+/K+ exchange, provides the concentration and osmotic forces required for net HCl secretion. Whether and how the K+ and Cl- pathways are activated by stimulation and/or how they get to the apical membrane domain remain important questions. With respect to mechanisms of parietal cell activation, secretagogue coupled elevation of cAMP and activation of protein kinase A form the basis of a well-established second messenger pathway. Several laboratories have identified various proteins that are phosphorylated concomitant with parietal cell stimulation, representing numerous candidates for effectors in stimulus-secretion coupling. Here, we emphasized the possible involvement of an 80-kDa protein whose phosphorylation was correlated with the cAMP pathway of HCl secretion. Immunocytolocalization of the 80-kDa phosphoprotein to the apical membrane and associated actin microfilaments prompted our suggestion that this protein might serve as a linkage between plasma membrane and cytoskeleton. Search for a possible role for the 80-kDa phosphoprotein in apical surface organization, stability, and turnover should represent an important thrust of research. Further understanding of the mechanism of cell activation will require a more complete elaboration of the functional role of many activation-related proteins. PMID- 2561318 TI - Regulation of CA II and H+,K(+)-ATPase gene expression in canine gastric parietal cells. AB - Acid secretagogue-specific receptor activation in parietal cells triggers rapid and coordinate gene expression of gastric H+,K(+)-ATPase and of CA II. The rapid rise in steady-state levels of CA II mRNA is due to new transcription of the CA II gene in stimulated cells. Although the presumed function of CA II in activated parietal cells is to catalyze the generation of HCO3- from OH-, regulation of CA II gene expression appears to be independent of the generation of H+ (and OH-) through the action of H+,K(+)-ATPase. PMID- 2561319 TI - Transmembrane segments of the P-type cation-transporting ATPases. A comparative study. AB - The transmembrane segments predicted for the Neurospora H-ATPase are laid out diagrammatically in Figure 10. Although the eight segments have arbitrarily been compressed into rectangles of the same size, they range in length from 20 residues (II) to 30 residues (IV and VI), so the corresponding helices must vary in length as well. Notable features of the model include the charged residues located just outside the plane of the membrane, with a clear excess of negative charges (5-, 1+) at the extracellular surface and a slight excess of positive charges (4+, 3-) at the cytoplasmic surface. There are also a conspicuous number of bulky residues (tryptophan, phenylalanine, and tyrosine) just inside the plane of the membrane. Within the bilayer, most of the helices are noticeably amphipathic, consistent with the expectation that at least some of them stack together to form a channel-like structure with a hydrophobic surface and a hydrophilic core. The charged residues predicted to lie within the membrane are listed in Table 2, which is a summary of data from eight of the P-type ATPases; the S. cerevisiae and S. pombe enzymes have not been included because they are nearly identical in this respect to the Neurospora enzyme. Interestingly, all of the ATPases have more membrane-embedded negative charges (5 to 8) than positive ones (0 to 4), a pattern that may be connected with their role as cation transporters. Certainly, other unrelated transport proteins have a rather different pattern of positive and negative charges: for example, the mammalian glucose transporter (1+, 2-), Na-glucose transporter (3+, 3-), and the E. coli lac permease (11+, 7-). The actual positioning of the negative charges in the P type ATPases does not make it easy to single out the functionally important ones, however. The glutamyl residue in segment I is present in the fungal, plant, and Leishmania H-ATPases but not in the gastric H,K-ATPase. The same is true for the aspartate in segment II, except that it also appears in the muscle and brain Ca ATPases. A glutamate is found at one end of segment III in the E. coli and fungal enzymes and at the other end in Arabidopsis; in segment IV, another glutamate appears in a well-conserved region in the Leishmania and mammalian enzymes but not in the bacterial, fungal, or plant ones.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2561320 TI - Models for gastric proton pump in light of the high oxyntic cell conductance. PMID- 2561321 TI - Differential function properties of a P-type ATPase/proton pump. PMID- 2561322 TI - Chloroplast thylakoid membrane-bound Ca2+ acts in a gating mechanism to regulate energy-coupled proton fluxes. PMID- 2561323 TI - Cell volume changes and the activity of the chloride conductance path. PMID- 2561324 TI - Possible role of outwardly rectifying anion channels in epithelial transport. PMID- 2561325 TI - Role of the cytoskeleton in regulation of gastric HCl secretion. PMID- 2561326 TI - Regulation of transepithelial chloride transport by amphibian gallbladder epithelium. PMID- 2561327 TI - Mechanisms of chloride transport in secretory epithelia. PMID- 2561328 TI - Characterization of the proton-secreting cell of the rabbit medullary collecting duct. PMID- 2561329 TI - Cl- conductance and acid secretion in the human sweat duct. PMID- 2561330 TI - Phosphorylation-dependent regulation of apical membrane chloride channels in normal and cystic fibrosis airway epithelium. AB - The observations described herein allow us to make several inferences about PKC and regulation of normal and CF Cl- channels. FIGURE 5 shows a model that summarizes these observations. In this model, for the sake of clarity, we refer to the channel as a single entity, but note that it may consist of multiple subunits and associated proteins. FIGURE 5A shows the channel in an inactivated state following excision from the cell. The channel can be activated by strong membrane depolarization, via an unknown mechanism, or by phosphorylation with PKA or PKC at a low [Ca2+] We speculate that PKA and PKC may phosphorylate and activate the channel at the same site, or region of the channel, because phosphorylation-dependent activation by both is defective in CF. This result suggests that the CF defect might lie in a defective phosphorylation site on the channel, or associated protein, or in the mechanism that converts phosphorylation into a change in channel conformation, such as activation. Activated channels can be inactivated by PKC at a high [Ca2+]. At high [Ca2+], PKC maintains the channel in an inactivated state and it inactivates channels that have been activated by PKC at low [Ca2+], by depolarization, or by PKA. Both activation and inactivation appear to result from phosphorylation; neither can be explained by down regulation of the channel. There are several possible ways to explain the two opposite effects of PKC on the Cl- channel: different responses may be due to an effect of Ca2+ on the channel, on PKC, or on the interaction between the two.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561331 TI - Acid and alkali secretion by the turtle urinary bladder. A model system for neurohormonal regulation of acid-base homeostasis. PMID- 2561332 TI - Protective effect of rhinovirus receptor blocking antibody in human fibroblast cells. AB - Infection of HeLa cells by human rhinoviruses (RV) of the major receptor group is inhibited by a HeLa-derived rhinovirus receptor murine monoclonal antibody (RRMA). In yield reduction studies in human embryonic lung fibroblast cells, pretreatment with 1.0 or 10 micrograms/ml of RRMA partially protected (greater than 90% titer reduction) against infection by RV 39 or coxsackie A21 (members of the major receptor family), but not by RV 1A (member of the minor receptor family). The protection afforded by RRMA persisted at least 72 h after a 2-h exposure. These results suggest that RV receptors can be effectively blocked for prolonged periods in cultured fibroblast cells. PMID- 2561333 TI - Evaluation of new antiviral agents: II. The use of animal models. AB - Antiviral chemotherapy has become a reality in the 1980s. Since the use of animal models in the testing of new antiviral agents is an inevitable step prior to clinical trial in human patients, it is important to understand the basic principles of using model systems. Briefly reviewed in this paper are the heterologous and homologous animal models which have been used for studies of various herpesvirus infections in humans. Discussions of the use of the guinea pig models mainly, for members of the Herpesviridae are presented in more detail. Precautions needed for the development of new animal models, and suggestions proposed for the use of animal models for testing new antiviral agents are outlined. It is hoped that new animal models will be developed in the foreseeable future for evaluating the much needed effective but less toxic antiviral agents for a variety of human viral diseases. PMID- 2561334 TI - Arenavirus infection in the guinea pig model: antiviral therapy with recombinant interferon-alpha, the immunomodulator CL246,738 and ribavirin. AB - Human arenaviral infections have a high mortality, and are dangerous to work with in the laboratory. There is a need for good antiviral agents to treat these infections. Pichinde virus infection of the inbred strain 13 guinea pig is a relatively safe, good animal model for human arenavirus infections. Mortality is consistently 100% between days 12 and 25 (mean 14.8) days after infection. When infected animals were treated with recombinant human interferon alpha A, or with CL246,783, an immunomodulator known to induce interferon, no beneficial effect was noted. When animals received ribavirin, 25 mg/kg/day for the first 14 days of infection, the course of infection was prolonged, with death occurring a mean of 22.5 days after infection. If ribavirin was administered for 28 days, mortality was reduced to 25%, with those animals dying a mean of 21.0 days after infection. These results confirm the studies that indicate that ribavirin is a useful agent for treating arenaviral infections. However, treatment with this agent must be prolonged. They also demonstrate the potential usefulness of Pichinde virus infection in strain 13 guinea pigs as an animal model of human disease. PMID- 2561335 TI - Local and systemic antibody response to rotavirus WC3 vaccine in adult volunteers. AB - An evaluation of the safety and immunogenicity of WC3 rotavirus vaccine was evaluated in adult volunteers. Pre- and post-vaccination titers of neutralizing antibody to WC3 and to the four human rotavirus serotypes as well as serum and stool rotavirus IgA levels were measured. Vaccination was safe and did not induce elevation of liver enzymes. None of the 12 volunteers receiving WC3 vaccine shed detectable amounts of virus although antibody rises were detected in 11 of 12 vaccines. Nine developed and increase in WC3 neutralizing antibody, one additional subject had a rise in Wa (human serotype 1) neutralizing antibody while another subject only developed a rise in stool rotavirus IgA. All of the vaccine recipients with a rise in WC3 neutralizing antibody also developed a rise in neutralizing antibody against at least one of the four most common human rotavirus serotypes. A stool IgA rotavirus antibody response was detected in 6 of 9 WC3 recipients with measurable stool antibody. None of the control subjects developed significant rises in any of the antibody titers measured. WC3 rotavirus vaccine appears to be safe and induces systemic and local immune responses in adults suggesting that further evaluation of WC3 should be considered in infants. PMID- 2561336 TI - Phosphonoacetic acid inhibits replication of human herpesvirus-6. AB - Phosphonoacetic acid (PAA) inhibits the replication of human herpesvirus-6 (HHV 6) in mononuclear cells from cord bloods which are susceptible for natural HHV-6 infection in humans. Nuclear extracts of uninfected or HHV-6-infected mononuclear cells were applied to phosphocellulose column chromatography, and DNA polymerase activity was measured with or without the addition of 100 mM ammonium sulfate. The major DNA polymerase activities eluted at 0.47 M KCl were suppressed in both uninfected and HHV-6 infected cells by the addition of 100 mM ammonium sulfate. DNA polymerase activity eluted at 0.47 M KCl was observed only from HHV-6 infected cells; it was enhanced by 100 mM ammonium sulfate and neutralized with immune serum. DNA polymerase activity eluted at 0.73 M KCl was determined to be HHV-6 specific and had the properties of a typical herpesvirus-induced DNA polymerase. PAA inhibited HHV-6-specific DNA polymerase activity. PMID- 2561337 TI - [The relationships between antitoxic titers determined in a group of young people before and after booster ADT by the in vivo neutralization test. Comparative NT PHAR evaluations]. AB - The geometrical mean of antitoxic titers determined in a lot of immunized young men who received, 5-7 years before, a booster with diphtheria-tetanus bivaccine, was of 1.54 (x/divided by 10.54) I.U./ml. At one month after another booster, performed with Tetanus toxoid (adsorbed), an increase of 34 times was recorded, the geometrical mean reaching the value of 52.67 (x/divided by 10.42 I.U./ml). The covering coefficients of the minimal protective antitoxin level have increased from 154 to 5267, in the estimation performed at the level of geometrical mean (calculating the ratio between the value of this indicator and that of the protection limit) and from 1623 to 54935, or from 15 to 505, respectively, in the estimations performed at the levels of the limits of the statistical range of one geometric standard deviation (Mg x/divided by D Sg). The study of the relations between pre- and post-antitoxic titers, performed by the linear regression analysis, leads to an equation whose slope evidenced the lower amplitude of the booster titers, along the increase of previous titers. The study of the inter-methods (neutralization-passive haemagglutination) regression allowed to obtain a correlation coefficient, between methods, of r = 0.83 and to perform the transposition of the minimal protective N.T. level (0.01 I.U.) in the terms of passive haemagglutination reaction, at the titer of 160 (0.3 PHAU/ml), for the p less than or equal to 0.05 probability level. PMID- 2561338 TI - [Clinico-pathologic correlations in 78 biopsies of the sural nerve]. AB - Peripheral nerve biopsies when processed with conventional techniques for paraffin embedding usually do not provide sufficient data for the diagnostic conclusion. However, if the nerve is processed for resin embedding for semi and ultra-thin sections and teasing of fibres, several aspects can be analysed including quantitative and morphometric data. We studied the sural nerve biopsy of 78 patients examined at the Antonio Pedro University Hospital, Niteroi RJ, applying those techniques and we found that in 55 cases (70.5%) the pathologic diagnosis was conclusive, in 11 (14.1%) although the nerve had abnormalities it was not possible to establish a diagnosis, and in 12 (15.4%) the nerve was normal. In 68 cases there was a clinical diagnosis which was confirmed in 49 but not in the remaining 19, since 8 had non-specific changes and 11 were normal. From the 10 cases which did not have a clinical diagnosis the biopsy was conclusive in 6, showed non-specific changes in 4, and was normal in 1 case. The pathologic conclusion in most of our cases was possible because not only we had the clinical data but all the nerves were processed for resin embedding. PMID- 2561339 TI - [Disseminated herpetic encephalitis: report of an autopsy case with immunohistochemical study]. AB - An autopsied case of disseminated herpetic encephalitis in a previously healthy one-year-and-three-month-old child is reported. He had fever, repeated convulsive crises and alterations of consciousness which progressed into a coma, leading to death in eight days. The neuropathological picture was characterized by necrosis and inflammation in multiple foci disseminated in the cerebral hemispheres, brainstem and cerebellum. The Avidin-Biotin-Peroxidase Complex technique showed, in neurons of the thalamus, intra-cytoplasmatic immunoreactivity, and, very rarely, intranuclear for the Herpes simplex virus type 1 antigens. This case differs from the other herpetic encephalitis ones described in the literature in two aspects: (1) by the dissemination of the lesions, in contrast with the forms topographically limited to the limbic system and, less commonly, to the brainstem; (2) by the presence of necrosis, inflammation and focal hemorrhage, which are alterations that practically do not exist in cases of disseminated encephalitis typically described in immuno-depressed individuals. PMID- 2561340 TI - [Myopathy due to succinate cytochrome C oxidoreductase deficiency: possible defect of complex II of the respiratory chain]. AB - The case of a 24 years-old woman with weakness since the teens and progressive loss of muscle strength is reported. The muscle biopsy showed increased number of mitochondria. In two occasions the respiratory chain enzymes showed important reduction of the succinate-cytochrome-C-reductase, suggesting a possible defect in the complex II of the respiratory chain. Large doses of vitamins C and K were prescribed. There was improvement of muscle strength. A discussion about the most common syndromes marked by mitochondrial abnormalities in muscle is made, as well as about the type of work-up that should be done in suspect cases of respiratory chain defects. PMID- 2561341 TI - [Benign fibrohistiocytoma of the soft palate]. AB - A case of benign fibrohistiocytoma of the soft palate in a 61 years-old woman is presented and current approaches to histogenesis of fibro-histiocytic tumors commented. PMID- 2561342 TI - Vaccines. PMID- 2561343 TI - Immunopathology of lentiviral infections in ungulate animals. AB - The immunopathogenesis of lentiviral lesions in sheep and goats requires continuous replication of the virus in tissues of the animal. This entails escape from various defense mechanisms of the host. Viral expression occurs mainly in tissue-specific macrophage populations and viral proteins produced by the cells induce and combine with antibodies to form immune complexes. These may be pathogenic locally. Infected macrophages also present lentiviral antigens to T lymphocytes and this results in a cascade of cellular responses including proliferation and accumulation of CD8 cells. Cytokines including interferon(s) are produced by lymphocytes and these enhance the antigen-presenting capacity of the macrophages. These lymphoproliferative cellular responses vary from those in human immunodeficiency virus- and simian immunodeficiency virus-infected hosts, mainly because CD4 cells of sheep and goats are not killed by the viruses. These cells, therefore, respond immunologically to viral antigens and this leads to active-chronic inflammation. PMID- 2561344 TI - The biology and immunopathology of simian immunodeficiency virus infection. PMID- 2561345 TI - Amino acid sequence and function of rubredoxin from Desulfovibrio vulgaris Miyazaki. AB - Rubredoxin was purified from Desulfovibrio vulgaris Miyazaki. It was sequenced and some of its properties determined. Rubredoxin is composed of 52 amino acids. It is highly homologous to that from D. vulgaris Hildenborough. Its N-terminal methionyl residue is partially formylated. The millimolar absorption coefficients of the rubredoxin at 489 nm and 280 nm are 8.1 and 18.5, respectively, and the standard redox potential is +5 mV, which is slightly higher than those of other rubredoxins. Rubredoxin, as well as cytochrome c-553, was reduced with lactate by the action of lactate dehydrogenase of this organism, and the reaction was stimulated with 2-methyl-1,4-naphthoquinone. It is suggested that rubredoxin, in collaboration with membranous quinone, functions as a natural electron carrier for cytoplasmic lactate dehydrogenase of this organism, whereas cytochrome c-553 plays the same role for periplasmic lactate dehydrogenase. PMID- 2561346 TI - Structural analysis of prokaryotic and eukaryotic 5S rRNAs by RNase H. AB - The availabilities of single-stranded 5S rRNA regions c, d and d' for base pairing interactions were analyzed by using synthetic DNA oligomers. Hybrid formation was detected by the endonucleolytical mode of the RNA-DNA specific action of RNase H. Provided that the hybrid interaction involved 6 successive base pairs, 5S rRNA loop c nucleotides 42-47 displayed accessibility in Escherichia coli, Bacillus stearothermophilus and Thermus thermophilus 5S rRNAs as well as in eukaryotic 5S rRNAs from Saccharomyces carlsbergensis, Rattus rattus and Equisetum arvense. Investigating eubacterial 5S rRNA regions d and d' (nucleotides 71-76 and 99-105, respectively), susceptibility was observed in E. coli 5S rRNA which, however, decreases in B. stearothermophilus and even more so in T. thermophilus 5S rRNA. For additional evaluation of the data obtained by RNase H cleavage, association constants of the hexanucleotides were determined by equilibrium dialysis at 4 degrees C for B. stearothermophilus 5S rRNA. The results obtained reveal that nucleotides 36-41 of B. stearothermophilus 5S rRNA are inaccessible for Watson-Crick interaction, which suggests that this part of loop c is in a structurally constrained configuration, or buried in the tertiary structure or involved in tertiary interactions. PMID- 2561347 TI - Murexide bleaching: a new direct assay method for characterizing reactive oxygen species. AB - The murexide (5,5'-nitrilodibarbituric acid, monoammonium salt) is an efficient scavenger for superoxide and hydroxyl radicals. When exposed to oxygen radicals, murexide is converted to a colorless alloxan derivative and its absorbance at 520 nm decreases in proportion to the radicals produced. It is used to detect these reactive oxygen species in biochemical systems such as acetaldehyde oxidation by xanthine oxidase and the respiratory burst of polymorphonuclear leukocytes induced by phorbol 12-myristate, 13-acetate. The method was sensitive enough to allow direct monitoring of the production of superoxides from 10(6) phorbol 12 myristate, 13-acetate polymorphonuclear leukocyte-stimulated cells. Moreover, murexide bleaching is inhibited in the presence of radical scavengers, allowing a comparison of their scavenging activities. PMID- 2561348 TI - [The blocking action of platelet-activating factor (PAF) on the calcium currents of the atrial fibers in the frog and guinea pig]. AB - The effects of platelet-activating factor (PAF) on the myocardial cell membrane Ca-current (ICa) and Ca-action potential (Ca-AP) were investigated. In double sucrose-gap voltage-clamped frog atrial trabeculae PAF (2 X 10(-7) M) reduced ICa amplitude to 40-50%; at the same time the IK-amplitude was increased to the same value. These changes of ICa and IK amplitudes were protected by simultaneous action of PAF and PAF antagonist BN 52021 (4 X 10(-6) M). In the partially depolarized (K+0 = 15-20 mM) of the guinea pig myocardial auricles PAF decreased Ca-AP amplitude and Vmax of its upstroke and shortened the Ca-AP duration (intracellular microelectrodes) like the isometric tension responses. These effects were prevented by PAF antagonist U-66985. Histamine was also able to protect from the PAF-induced changes of Ca-AP and tension responses. Our data demonstrated both by direct and by indirect methods of ICa registration in myocardia membrane that PAF induces reversed blocking of ICa. Because the blocking effects of PAF on frog and guinea pig myocardium are identical, these results imply that the mechanisms of PAF action on cold- and warm-blooded animals are similar in principle. The coupling of ICa and IK changes confirm our earlier supposition that PAF-induced Ca-AP shorting can be explained by IK augmentation. PMID- 2561349 TI - [The development of desensitization during rhythmic activity of the synapse and in the course of generating a single signal]. AB - The possibility of desensitization (DS) development under the rhythmic activity and during the generation of the single response to acetylcholine was studied in the frog sartorius muscle by voltage-clamp technique. It was revealed, that under conditions promoting DS (hyperpolarization, muscle warming, rise of calcium concentration as well as treatment by the DS--potentiating agent--proadifen) decrease of the postsynaptic membrane sensitivity to the acetylcholine can develop after few multiquantal end-plate currents--and when acetylcholinesterase is inhibited--during the response to a single quantum of acetylcholine. PMID- 2561350 TI - [Histological study of hydroxylapatite mixed with collagen as osseous implant in mandible]. AB - By using five Taiwan Macaca monkeys as experimental animals, three bony defects about 5 x 5 x 3 mm in size were created at each of their buccal region of mandible. Bovine skin collagen and particulate hydroxylapatite were used as implant materials. The artificial defects implanted with hydroxylapatite/collagen mixture were experimental group and the defects implanted with collagen or without any implantation were control groups. The monkeys were sacrificed at the time of 1,2,3,6,9 months after implantation surgery and were injected with tetracycline according to the above intervals at one week before being sacrificed. The specimens were divided into two parts, one of which was decalcified and stained with hemotoxylineeiosin for observation under optic light microscope, the other without decalcification was made into ground section for observation under fluorescent microscope. The result revealed that the healing processes of the two control groups were similar with each other. Bone formation was delayed in HA/Collagen group because it took almost five months to achieve complete bony repair of the mandibular defect. However bone formation at the HA/Collagen implant site was earlier than that at the pure HA implant site in our similar study. What's the role of collagen that played during bone formation still needed further investigation. The tissue response was good without obvious inflammatory cells infiltration. Clinical application of block form of dried HA/Collagen mixture was suggested for it was easier to handle and could decrease the flow of HA particles after implantation. PMID- 2561351 TI - [Malignant fibrous histiocytoma of spleen. A case report and literature review]. AB - A 11 year old girl who presented with malaise, poor appetite and flank mass was admitted in Sept. 1988. Abdominal x-ray, sonogram, Gallium scan and CT scan revealed a tumor mass in the spleen. Laparotomy confirmed a hugh mass measuring 15x15x10 cm in the spleen. The histologic study proved the tumor to be malignant fibrous histiocytoma, inflammatory type. Malignant fibrous histiocytoma occur mainly in late adult life, we report this case because the patient is young and the involvement of the spleen is rare. PMID- 2561352 TI - Low levels of mumps virus antigen induced interferon-alpha production in insulin dependent diabetes. AB - The in vitro production of interferon-alpha was studied in cultures of peripheral blood mononuclear cells from children with insulin-dependent diabetes. Significantly lower levels (p less than 0.01) of interferon (median 64 IU/ml) were found in mumps antigen stimulated cultures of IDDM patients compared to control children (median 256 IU/ml) whereas no differences between groups were found in the amount of interferon induced by Coxsackie B pool antigen or live Sendai virus. PMID- 2561354 TI - Communicable disease report. April to June 1989. The PHLS Communicable Disease Surveillance Centre. PMID- 2561353 TI - Antioxidant enzyme status and lipid peroxidation in various tissues of diabetic and starved rats. AB - The effect of short term (2-wk) diabetes induced by streptozotocin and starvation (1-wk) on antioxidant enzymes and lipid peroxidation in the liver, kidney and heart of rats was investigated. The activity of mitochondrial oxidative markers was increased in diabetic liver and kidney, while the activity in tissues of starved rats tended to be decreased. Immunoreactive manganese superoxide dismutase was increased only in diabetic liver and was unchanged or decreased in the rest of the tissues. Glutathione peroxidase activity was increased in tissues of diabetic but not starved rats. The changes in copper-zinc superoxide dismutase and catalase in diabetic rats were similar to those in starved rats. In both groups, copper-zinc superoxide dismutase was decreased in liver, while catalase activity was decreased in liver and kidney, and increased in heart. The lipid peroxide level was increased in diabetic kidney and in the heart of starved rats, and decreased in the rest of the tissues. Insulin treatment in diabetic rats and refeeding in starved rats restored most of the abnormalities toward normal. These results suggest that accelerated mitochondrial oxidative metabolism not accompanied by induction of manganes superoxide dismutase results in oxidative injury in the hypertrophied kidney at an early stage of diabetes and possibly contributes to the development of nephropathy. Peroxidative myocardial damage in starved rat appears to be mediated by a catabolic process. PMID- 2561355 TI - [Metastatic cancer in the ovary--report of 57 cases]. AB - Fifty-seven patients with metastatic ovarian cancer arising from extra-genital sites are analysed. 80.8%, 14% and 5.3% of the primary cancers were from gastrointestinal tract (GI), breast and lung, respectively. The primary carcinoma was diagnosed before the discovery of ovarian tumor in 24 patients. The ovarian and primary carcinomas were found simultaneously in 11 patients (including one autopsy), while in 22 patients, the primary cancer was not discovered until the ovarian tumor had been resected. Both ovaries were involved in 60% of GI cancer and 12.5% of breast cancer. 91.2% of the patients were found to have metastasis to other tissues and organs outside the ovary. Of patients with GI cancer, 80.8% had metastasis to the abdomino-pelvic cavity, 73.7% had ascites and 52.2% had lymphatic metastasis. The majority of the patients with breast cancer had lymphatic (75%) and hematogenous (50%) metastases. In this series, the prognosis was poor in all the patients with an average survival time of 11 months. 63.2% and 86% of the patients died within 1 and 2 years. These facts indicate that ovarian involvement may be an important part of widespread dissemination from the primary cancer. PMID- 2561356 TI - Protein-DNA interactions at steroid hormone regulated genes. AB - This work summarizes binding data that were obtained with partially purified glucocorticoid and progesterone receptors, as well as with a crude nuclear protein extract, to DNA sequences in and around hormonally regulated genes. The sequence recognition by the glucocorticoid receptor at the different defined glucocorticoid regulatory elements (GRE) is discussed and a consensus sequence formulated. A three dimensional representation gives an impression of the mode of interaction between the protein and the double helix of DNA. In the promoters of mouse mammary tomour virus (MTV) and chicken lysozyme overlapping binding sites for both, glucocorticoid and progestine-receptors are found that are responsible for the hormonal inducibility of the genes. In crude extract from rat liver nuclei, a nonhistone protein is found that specifically binds to sequences on the MTV-LTR region overlapping the GRE. The possible implication of this protein in hormonal regulation of transcription is discussed. PMID- 2561357 TI - [Advantages of a cardiologic ambulatory care facility specifically organized for patients undergoing treatment with oral anticoagulant drugs]. AB - In patients with artificial heart valve prosthesis oral anticoagulants reduce but not eliminate the thromboembolic complications however, they do increase the risk of bleeding. In the present study, the incidence of thromboembolic and hemorrhagic complications in two homogeneous groups of patients with artificial heart valves on long term oral anticoagulant treatment has been evaluated. Group A (99 patients; total follow-up = 309 years) were resident in the Triveneto regions and received a questionnaire while group B (104 patients; total follow-up = 370 years) were referred to our department's centre for the control of oral anticoagulant treatment. Both groups were kept at a therapeutic range of 20-30% in terms of prothrombin activity. The incidence of thromboembolic and hemorrhagic complications is expressed as the number of episodes per 100 patient/years. Thromboembolic episodes were 2.6 (1.3 fatal) in group A while they were 1.08 (0.27 fatal) in group B; the reduction of fatal thromboembolic events was statistically significant (p less than 0.05). Hemorrhagic episodes were 1.9 (0.63 fatal) in group A while they were 0.81 (0 fatal) in group B. We concluded that an organized control of oral anticoagulant treatment in patients with artificial heart valves is advantageous as it significantly reduces fatal thromboembolism. Moreover, it could reduce the incidence of total thromboembolic and hemorrhagic episodes by more than 50%. PMID- 2561358 TI - [Localization in the body of the mobile element MDG1 of 2 regions specifically binding with Drosophila nuclear proteins]. AB - Two regions in mdg1 mobile element's body can specifically bind nuclear proteins of Drosophila melanogaster, as demonstrated by the method of retention of DNA protein complexes of nitrocellulose filters. The first region is situated in the 5'-end part of mdg1, 1 kb downstream the site of initiation of transcription and contains long oligo (A) blocks (from 14 to 30 nucleotides) in the coding chain. The second region is localized near the 3'-LTR and consists of tandem 14 nucleotide repeats and a palindrome, destruction of which leads to weaker binding. There is no competition between the two regions for proteins, which evidence that they are recognized by the different proteins. The binding with the first region can be suppressed by adding the 412 mobile element DNA. These regions are supposed to take place in the regulation of mdg1 transcription. PMID- 2561359 TI - [The role of gonadotropins, cyclic AMP, 22-R-hydroxycholesterol and cofactors in regulating endocrine functions of the Leydig cells in rats. II. Effects of LH, hCG, dbcAMP, 22-R-OH-cholesterol and cofactors on the synthesis of pregnenolone and testosterone in the interstitial gland of rats]. AB - The authors examined in vitro the influence of gonadotrophins, cAMP, 22-R-OH cholesterol and cofactors on the synthesis of pregnenolon and testosterone in the interstitial gland of the rat. Sections of the nucleus were incubated with LH (100 ng/ml), hCG (1.0 j.m./ml), dbcAMP (1 mM), 22-R-OH-cholesterol (30 microM) and cofactors (NAD + NADP + G-6-P + G-6-PDH). It was found that an increase in concentrations of hCG above physiological values was not accompanied by an increase in secreting steroid hormones. LH, hCG and dbcAMP increased the synthesis of pregnenolon twice, and testosterone--three times. 22-R-OH cholesterol as a substrate increased the synthesis three and four times respectively, and added cofactors five times and four and a half times respectively. Joining 22-R-OH-cholesterol or a cofactor with LH does not intensify a stimulating effect. PMID- 2561360 TI - [The role of gonadotropins, cyclic AMP, 22-R-OH-cholesterol and cofactors in regulating endocrine functions of the Leydig cells in rats. III. Mechanisms responsible for "desensitization" of the Leydig cells of rats caused by high doses of hCG]. AB - Two groups of rats (a control group and the group examined) were administered intraperitoneally supraphysiological doses of hCG in order to induce a "down regulation" effect on the level of receptors LH and to achieve the desensibilization of Leydig cells. The authors tried to find out at which stage of sequence of changes from receptor stimulation to hormone production there appears a state of cellular resistance to further stimulation. Sections of the nucleus were incubated with various substances influencing steridogenesis (LH, hCG, dbcAMP, 22-R-OH-cholesterol, NAD + NADP + G-6-P + G-6-PDH). An index of the influence of the above substances on the synthesis of androgens were amounts of pregnenolon as the first and testosterone as the final stage of hormonal changes marked radioimmunologically in nucleus homogenates and incubating media. It was shown that the resistance of Leydig cells to further stimulation in the group of animals that were given high doses of hCG is the result of enzymatic blocks in testosterone synthesis. The first block is "late" block of 17 alpha-hydroxylase and 17-20 desmolase, disturbing transforming of 21-carbon steriods into 19-carbon androgens. When the dose of hCG increases, there appears the second block, the so called "early" block, disturbing mitochondrial synthesis of pregnenolon. It was found that exogenic cofactors are in a position, at least partially, to restore the activity of blocked enzymes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561361 TI - Non-proteolytic activation of latent human neutrophil collagenase and its role in matrix destruction in periodontal diseases. AB - Collagenases are known to be associated with tissue destruction in chronic inflammatory diseases such as periodontal diseases and rheumatoid arthritis. Collagenases are secreted by circulating inflammatory cells (polymorphonuclear leukocytes and monocytes), resident mesenchymal cells and epithelial cells in latent forms, which can be activated by proteases and compounds reacting with protein thiol groups. We have studied here the effects of oxygen-derived free radicals (ODFR) on latent human neutrophil collagenase. Also, in order to elucidate the cellular sources of collagenases, the ability of human gingival crevicular fluid (GCF) collagenases both from adult periodontitis (AP) and localized juvenile periodontitis (LJP) patients to degrade soluble interstitial collagen types I and II was studied. ODFR generated by the xanthine oxidase/hypoxanthine system in the presence of trace amounts of iron and EDTA activated latent neutrophil collagenase to an equal extent as the known activators phenylmercuric chloride and gold thioglucose. ODFR activation was inhibited by desferoxamine and mannitol as well as by superoxide dismutase and catalase. Clear differences in the susceptibility of collagen types I and II to AP and LJP GCF collagenases were observed. AP GCF collagenase degraded type I and II collagens at equal rates, resembling the substrate-specificity of human neutrophil collagenase. LJP GCF collagenase degraded type I collagen considerably faster than type II collagen, which was only negligibly degraded. This corresponds to the substrate specificity of fibroblast collagenase. Zymographic evaluation of gelatinolytic proteases showed the presence of 90 and 68 kD gelatinases in both AP and LJP GCF. Non-proteolytic means apparently provide a potent activation pathway of neutrophil collagenase in vivo and the hydroxyl radical was identified to be one of the potent activating oxidants.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561362 TI - Role of Salmonella enteritidis lipopolysaccharide on anti-HSV activity of macrophages from different anatomical sites. AB - It is generally agreed that endotoxins from Gram-negative bacteria play a modulatory role on several macrophage functions. The intrinsic activity versus herpes simplex virus type 1 and type 2 of Kupffer cells, peritoneal and alveolar macrophages, harvested from normal and tumour-bearing rats, was evaluated. Moreover, the effects of different intravenous treatments with S. enteritidis endotoxin were investigated. The antiviral activity of peritoneal, alveolar macrophages and Kupffer cells from tumour-bearing rats is definitely impaired but it appears to be positively modulated by in-vivo administration of S. enteritidis lipopolysaccharide (LPS). PMID- 2561363 TI - Glomus tumours of the hand. AB - Based on 8 cases of glomus tumour of the hand, observed over the last 3 years, the authors review the clinical features and pathological anatomy of this rare condition. They propose nuclear magnetic resonance as a preoperative instrumental study as an alternative to angiography because it is non-invasive, free from risk and is clinically very effective. PMID- 2561364 TI - Two-dimensional crystalline arrays of Na,K-ATPase with new subunit interactions induced by cobalt-tetrammine-ATP. AB - Purified membrane-bound Na,K-ATPase incubated with cobalt-tetrammine-ATP [Co(NH3)4ATP], which is a stable MgATP complex analog, shows two new types of membrane crystals, a new p21 form and a p4 form. The building blocks of the crystalline arrays correspond to (alpha beta)2 dimers of the enzyme protein suggesting that alpha-alpha interaction may be important in the pumping process. PMID- 2561365 TI - Intramitochondrial helical filaments in medullary tubules of the rat kidney. AB - Characteristic helical filaments were frequently found within dilated intracristal spaces of the mitochondria in epithelial cells of renal medullary tubules of normal rats of both sexes. These filaments had a helical structure with right-handed rotation. The approximate dimensions were 4 nm in thickness, 13 nm in helical diameter, and 16 nm in pitch. The filaments were common in all strains of rats examined in the present study but not in animals of other species including mice, guinea pigs, golden hamsters, Mongolian gerbils, house musk shrews, and rabbits. In Sprague-Dawley strain rats, the filaments were found not only in animals of all ages but also in 6-week-old germ-free animals and fetuses at the 18th day of gestation. Even in the same rats, the helical filaments were completely absent in the cortical tubules. PMID- 2561366 TI - Cytomegalovirus interstitial pneumonitis in a bone marrow transplant recipient. AB - A 15-year-old girl suffering from acute lymphoblastic leukemia developed cytomegalovirus interstitial pneumonitis 34 days after an allogenic bone marrow transplantation. The disease was diagnosed by open lung biopsy. Histopathologic examination disclosed intranuclear and intracytoplasmic inclusion bodies, as well as, a positive cytomegalovirus antigen detected by an immunofluorescence stain using a cytomegalovirus monoclonal antibody. The virus culture also eventually produced cytomegalovirus. Because of the lack of ganciclovir in this country, antiviral therapy of a non-specific nature was given to this patient. However, the treatment was ineffective and she subsequently died. There is an association between the immunologic events of a graft-versus-host disease and the development of cytomegalovirus interstitial pneumonitis. The pathogenesis of cytomegalovirus interstitial pneumonitis in a bone marrow transplant recipient is evidence of an immunopathologic disease, rather than that of a purely infectionus disease. years, the treatment modality of cytomegalovirus interstitial pneumonitis in bone marrow transplant recipients has become a combination of specific antiviral and immune therapy. PMID- 2561367 TI - Effects of the yeast extract components pyrroloquinoline quinone and aspartic acid on vitamin B12 production in Klebsiella pneumoniae IFO 13541. AB - The production of vitamin B12 from carbohydrates, peptone, casamino acid, etc., by intestinal bacteria was investigated. Klebsiella pneumoniae IFO 13541 was the most efficient strain for vitamin B12 production, which depended exclusively on the concentration of yeast extract added to the medium. A concentrated solution of yeast extract (1 ml) was chromatographed on a Sephadex G-25 column (1 x 180 cm) and eluted with H2O (eighty fractions of 3 ml each were collected). It was found that fractions in which bacterial growth was most prevalent also exhibited the highest amount of vitamin production. The effectiveness of yeast extract was shown by the participation of pyrroloquinoline quinone and aspartic acid in the growth stimulation and in the vitamin B12 production in this strain. PMID- 2561368 TI - [Classification of peripheral neuropathies]. PMID- 2561369 TI - [Peripheral neuropathies (P.N.) in a medical toxicology center--results obtained by a sample of 120 cases, of presumed toxic origin]. PMID- 2561370 TI - [A rare case of severe toxic peripheral neuropathy: poisoning by podophyllin. Apropos of 1 case]. PMID- 2561371 TI - [Acute arsenical peripheral neuropathy]. PMID- 2561372 TI - A scanning electron microscopic study of in vitro toxicity of ethylene-oxide sterilized bone repair materials. AB - Polylactic acid (PLA) and polyglycolic acid (PGA) have been under investigation for use in the management of hard- and soft-tissue wounds. Current research has included the incorporation of osteo-inductive substances into a PLA-PGA copolymer alloplastic implant material for enhancement of the healing of osseous defects. Conventional methods of sterilization--such as dry heat, steam heat, or 60Co- tend either to destroy or attenuate osteo-inductive activity and alter polymer biodegradation. Ethylene oxide (EO) gas sterilization is currently being tested as an alternate method. This study examined the relationship of EO-induced cytotoxicity to the length of time of polymer aeration following EO sterilization. Three groups of copolymer implant discs were studied: (1) 50:50 PLA-PGA copolymer, (2) PLA-PGA polymer with hydroxyapatite (HA), and (3) PLA-PGA with autolyzed, antigen-extracted (AA) bone particles. Polymer discs, as well as particulate HA and AA bone controls, were sterilized with EO for 12 hours. Following periods of two weeks, one week, one day, or no subsequent vacuum aeration, samples were placed into 24-well culture plates. A suspension of human fibroblasts was added to each well. Cell growth and attachment were permitted for 24 hours. Medium was then removed, and solutions for cell fixation, buffer washing, and dehydration were added to each well. SEM examination revealed changes in cell growth with increasing periods of aeration suggestive of increasing cell vitality. Cells growing on discs having no aeration were small, round, and lobulated, whereas those of seven to 14 days' aeration were more numerous, and flattened with many microvilli, pseudopodia, and dendritic processes, features consistent with normal cell morphology. These results suggest that EO-sterilized polymer implants should be aerated for least seven to 14 days prior to surgical use. PMID- 2561373 TI - The use of CT Multiplanar imaging to verify hydroxylapatite particle migration. PMID- 2561374 TI - [The immunoscintigraphic diagnosis of the metastases of colorectal cancer by using monoclonal antibodies]. AB - Sixty-four patients operated for colorectal cancer comprise the study group. CEA and CA-19.9 markers were used. The antibodies were labelled with 131I, using for the scintigraphy 99mTc as well. Eighty eight metastatic foci were detected, 67 of them (76.1 per cent) confirmed by the surgeon: Of 55 pelvic localizations, surgically detected were 47 (85.5 per cent), of 24 in the abdominal cavity 15 (62.5 per cent) were confirmed and of 9 in the liver--only 5 (55 per cent). Radioimmunoassay more accurate in pelvic localizations, whereas echography and computer tomography were more precise in liver examination (83.3 per cent). PMID- 2561375 TI - Neurological disorder in transgenic mice that express the large T antigen of polyoma virus in the nervous system. AB - Among a series of 44 transgenic families established after microinjection into fertilized eggs of a plasmid DNA where the structural gene for the large T antigen of polyoma virus is located downstream from the viral early promoter enhancer region, one family with a hereditary neurological disorder was observed. At about three weeks of age, these animals developed a syndrome of constant tremor with recurrent seizures. Histological and ultra-structural examination revealed extensive dysmyelination in the white matter of the brain stem, cerebellum and spinal cord, as well as of peripheral nerves. This phenotype is reminiscent of that of the mouse "twitcher" (twi) mutant and of the human hereditary leukodystrophies. Expression of the viral sequences, assayed by Northern analysis and immunolabeling of T antigen, occurred predominantly in cells of the central nervous system. Integration of the transgene was mapped by in situ hybridization on metaphasic plaques in region B-C1 of chromosome 12 (where the twi locus was previously localized). Long-term cultures of cells with neural characteristics could be established readily from the brain of the transgenic mice. PMID- 2561376 TI - [Escherichia coli phage receptors. Minor porins and proteins participating in the specific transport as phage receptors]. AB - Except for the main porin proteins OmpC and OmpF there exist the membrane proteins participating in the transport of specific substrates: phosphates, nucleosides, iron, vitamin B12, maltose and maltodextrins, that also play the role of phage receptors. Some phages use as receptors the porins determined by the genes of lambdoid prophages. LamB protein that serves receptor for phage lambda exposes the amino acids sequence on the outer surface of membranes that participates in phage adsorption. The sequence is similar to tetrapeptide of fibronectin responsible for binding with the surface of cellular receptor in eucaryotes. PMID- 2561377 TI - [Restriction analysis of the DNA of aleutian mink disease virus strain P-1]. AB - The II-1 strain of the Aleutian disease virus (ADV-II-1) was isolated from experimentally infected mink organs. The viral particles were isolated having 23 to 24 nm in diameter with the buoyant density of the virions in CsCl gradient being 1.41 g.ml-1. The single stranded ADV DNA extracted from the purified virus particles had the molecular mass about 1.4 . 10(6) (4800 bases). The double stranded replicative form of ADV DNA has been synthesized in vitro with the use of a large "Klenow" fragment of DNA-polymerase I. A restriction endonuclease map of ADV-II-1 DNA has been constructed with the use of in vitro synthesized double stranded DNA. PMID- 2561378 TI - [Primary structure of the gene for VP1 protein of the foot-and-mouth disease virus of Asia 1 serotype]. AB - The nucleotide sequence of the cDNA for the viral RNA region coding for the main antigenic protein of the epidemic stomatitis virus of Asia 1 serotype has been identified. The amino acid sequences in the regions of VP1 protein antigenic determinants of the serotype Asia 1 virus and other serotypes viruses have been compared. PMID- 2561379 TI - [Late results of the treatment of the chronic progressive form of multiple sclerosis by intensive immunosuppression]. AB - A group of 16 patients with secondary chronic progression of multiple sclerosis treated by short-lasting intensive immunosuppression (cyclophosphamide with ACTH) by the programme of Hausser et al (4) was compared with a clinically similar group of 16 patients treated exclusively with intramuscular ACTH. During 2 years of observation of the patients it was observed that short-lasting intensive immunosuppression induced with simultaneous administration of cyclophosphamide and ACTH had no beneficial effect on the slowly progressing form of multiple sclerosis. The observation leads to the conclusion that cyclophosphamide therapy should not be used in view of absence of favourable effect on the course of multiple sclerosis and the risk of increased frequency of malignant neoplasms which cannot be ruled out in this method. PMID- 2561380 TI - The effect of repeated administration of imipramine, citalopram and mianserin on responsiveness of central serotonergic, alpha 2-adrenergic and cholinergic system in mice. AB - The effect of antidepressant drugs: imipramine, citalopram and mianserin given either in a single dose or twice a day for 14 days in a dose of 10 mg/kg was investigated in mice in tests for the responsiveness of central serotonergic (L-5 hydroxytryptophan-induced head twitches), alpha 2-adrenergic (donidine hypoactivity) and cholinergic (oxotremorine syndrome) systems. The effect of L-5 hydroxytryptophan was inhibited by repeated administration of citalopram and mianserin but unchanged by administration of imipramine. After a single administration only mianserin inhibited the L-5 hydroxytryptophan effect. Given repeatedly, the investigated antidepressant drugs did not affect the effect of clonidine; only mianserin potentiated the hypoactivity when given in a single dose. Repeatedly administered antidepressant drugs did not affect the action of oxotremorine, although imipramine (but not citalopram or mianserin) antagonized it after a single administration. The results indicate that under the present conditions repeatedly given mianserin and citalopram, but not imipramine, antagonize behavioral effects of L-5 hydroxytryptophan. No one of the investigated antidepressant drugs given repeatedly changed the responsiveness of alpha 2 -adrenergic and cholinergic systems to their agonists. It might be concluded that the changes in alpha 1-adrenergic and dopaminergic systems, observed previously after repeated administration of antidepressant drugs, are selective. PMID- 2561381 TI - Stereospecific effects of (-)- and (+)-7-hydroxy-delta-6-tetrahydrocannabinol dimethylheptyl on the immune system of mice. AB - The effects of the (+) and (-) cannabinoid enantiomers 7-hydroxy-delta-6-tetra hydrocannabinol-dimethylheptyl [(-)-7-OH-delta-6-THC-DMH (HU-210) and (+)-7-OH delta-6-THC-DMH (HU-211)] on the inductive and productive phases of the primary humoral immune response to sheep red blood cell immunization were investigated in mice. Animals treated with (-)-7-OH-delta-6-THC-DMH (0.01, 0.05, 0.1 and 0.5 mg/kg) exhibited a dose-dependent suppression of both phases of the primary humoral immune response in the hemolytic plaque assay, the hemagglutination titer and in the ratio of the spleen weight to final body weight. Mice treated with (+) 7-OH-delta-6-THC-DMH (0.01, 0.05, 0.01, 0.5 and 1.0 mg/kg) did not exhibit dose dependent immune suppression. However, the (+) enantiomer reduced the number of plaque-forming cells in the hemolytic plaque assay during the inductive phase. Mice treated with a combination of (-) and (+) enantiomers, each at 0.01 mg/kg, exhibited no impairment in ability to undergo a primary humoral immune response (inductive phase). PMID- 2561383 TI - [Peripheral neuropathies in individuals infected with the human acquired immunodeficiency virus (HIV)]. PMID- 2561382 TI - [Rotavirus in the feces of children with acute diarrhea in Cochabamba, Bolivia]. AB - Eighty patients with acute diarrheal disease from Cochabamba, Bolivia, were investigated for rotavirus infection by rotapheresis. Rotavirus ARN was detected in 18 (22.5%) of the cases, thus suggesting that this agent is also a frequent cause of acute diarrhea in the studied population as reported from other places of the world. PMID- 2561384 TI - Separation of the sodium and potassium currents from an action potential of a single active fiber. AB - A method for calculation of the sodium and potassium currents of an active fiber is presented. The method includes the determination of the transmembrane ionic current, the sodium Nernst potential and the sodium conductivity as well. The method was illustrated by using an action potential of a single striated frog muscle fiber. The derivatives of the experimentally recorded action potential, necessary for calculation of the ionic current, were obtained by means of Fourier transform. PMID- 2561385 TI - Modification of single sodium channels in neuroblastoma cells by chloramine-T. AB - We have studied the behaviour of single sodium channels in cell-attached patches in neuroblastoma cell line N1E 115 at 8-15 degrees C modified by chloramine-T (CT). Application of the agent to the external surface of the membrane cause different effects: CT binds to the membrane and cause irreversible effects leading to a decrease of open channel lifetime at 10 or 20 mV relative potentials without affecting the amplitude of single channel current. Channels modified by CT show a tendency to occur in burst (they can open and close several times during depolarization). Therefore, the slower decay of the average current must be explained by the flickering behaviour of the modified channels. We also examined the effect of CT on the relationship between open time of the channels and membrane potential. On the other hand, a small decrease in the mean delay time as a function of membrane voltages was observed which probably is related to the three state model of inactivation. Our results suggest that CT modifies sodium channel inactivation. PMID- 2561386 TI - [The role of alpha adrenergic system in the regulation of pulpal circulation]. AB - Pulpal circulation is microcirculation, whose primary function is to transport the nutritive and return the waste metabolic products. As pulp is surrounded by a solid dentin sheath, proper regulation of its circulation, and of intra- and extracellular fluid levels is of utmost importance for the normal function of the pulp in physiologic conditions as well as for its survival in pathologic states. The alpha adrenergic system with all its components plays a key role in this regulation. PMID- 2561387 TI - [Low molecular weight heparins]. AB - The apparition in the 80's of low molecular weight heparins (LMWHs) obtained by chemical or enzymatical splitting, modified the fields of prophylactic treatment of thromboembolic diseases. These drugs present on heparin numerous advantages: equal antithrombotic activity with a smaller bleeding risk, higher bioavailability, longer pharmacokinetic, which simplify their use. The definite mode of action remains unknown, but LMWH act at different steps like coagulation (by AT III), fibrinolysis, blood cells, endothelial cells. Further, like heparin, they can be neutralized by protamine. However the right, simple, specific biological assay which presents a good correlation with the antithrombotic activity, remained to be established. PMID- 2561388 TI - Calcium binding proteins in the brain. AB - It is astonishing how a single ion, Ca++, can give rise to so many intracellular effects. The current explanation is that various combinations and permutations of a repertoire of calcium channels, calcium binding proteins and calcium pumps make up the individual answer of each neuronal type to the calcium signal. This review describes the major calcium binding proteins of the brain, which represent intracellular tools to decipher and transform the quantitative Ca+(+)-signal in qualitatively different cellular responses. PMID- 2561389 TI - Interrelationship between cyclic AMP level and chondrogenesis in vitro. AB - A consistent chondrogenesis takes place in micro-mass cultures of stage 23-24 chicken limb bud mesenchymal cells. In these cultures a short, marked elevation of cAMP level was detected at the time of the onset of cartilage phenotype expression. On the other hand, exogeneous glycosaminoglycans which inhibited chondrogenesis caused a reduction in the cAMP level of the cells. These correlations between cAMP level and phenotypic characteristics suggest that, among other things required in chondrogenesis, cAMP level may be a prominent factor. PMID- 2561390 TI - Inhibition of the semiconservative DNA synthesis and potentiation of the unscheduled DNA synthesis by 5-nitroimidazole derivatives with an imidazolium or pyridinium ring. AB - The effects of four pyridinium and one imidazolium derivatives of 5 nitroimidazole have been tested for their action on the kinetics of DNA semiconservative synthesis and of the unscheduled DNA synthesis carried out by TC SV40 cells. Unscheduled synthesis was induced by 25 Gy X-rays, and the kinetics were evaluated through the measurement of the newly synthetized DNA labelled with [3H-methyl]thymidine. All derivatives inhibit the DNA semiconservative synthesis and increase the unscheduled DNA synthesis to different degrees, but not in the suitable way to reveal the mechanism of interaction with DNA as responsible of the product's radiosensitizing actions on the same mammalian tumor cells. PMID- 2561391 TI - Low density lipoprotein binding by macroporous bead cellulose. AB - Cellulose of a certain macroporous structure is capable of binding selectively low density lipoproteins (LDL) whilst maintaining high density lipoprotein (HDL) levels. It was ascertained that the porous structure of the cellulose causes binding of LDL and that also diffusion processes play a role. To a certain extent special bindings such as hydrogen bonds between cellulose and LDL and/or hydrophobic interactions may furthermore be of importance for the LDL fixation. PMID- 2561392 TI - [Detection of human papillomavirus antigens in papillomatous lesions of the palate]. AB - The presence of human papillomavirus antigen was studied in palate papillomas and in verruca vulgaris located in the palate using the Peroxidase-Antiperoxidase immunohistochemical methods. The antigens were detected only in the nuclei of cells of the stratum granulosum and stratum corneum in some cases of verruca vulgaris but not in any case of palate papillomas. PMID- 2561393 TI - Rat brain capillary endothelial cells express functional PDGF B-type receptors. AB - Immunohistochemical staining revealed the presence of platelet-derived growth factor (PDGF) B-type receptors on capillaries of normal rat brain. Furthermore, capillary endothelial cells isolated from rat brain and grown in tissue culture bound [125I]PDGF-BB but not [125I]PDGF-AA, suggesting that they expressed B-type, but not A-type, PDGF receptors. PDGF-BB and PDGF-AB, but not PDGF-AA, also stimulated incorporation of [3H]thymidine into these cells. Thus, rat brain capillary endothelial cells have functional B-type receptors, and thereby differ from endothelial cells derived from large blood vessels, that do not express PDGF receptors. Our data suggest a possible role for PDGF-BB as an angiogenic factor. PMID- 2561394 TI - Endothelin is a potent constrictor of human intracranial arteries and veins. AB - Endothelin, a peptide produced in endothelial cells, was found to be a potent constrictor of human pial arteries through a direct, slow action on the smooth muscle cell, with a pD2 value of 8.54. It was less potent as a constrictor in pial veins (pD2 7.80) and in branches of the middle meningeal artery and superficial temporal artery (pD2 7.61 and 7.77). The major component of the contractile response in the arteries comprises an effect on potential-operated calcium channels, as evidenced by tests with nimodipine. Endothelin may regulate intracranial vessels tonically during physiological as well as pathophysiological conditions. PMID- 2561395 TI - Slow postsynaptic potentials recorded from hamster submandibular ganglion cells. PMID- 2561396 TI - Effect of aldose reductase inhibitor (Ponalrestat) on erythrocyte Na,K-ATPase activity in non-insulin-dependent diabetic patients with polyneuropathy. AB - The binding capacity of ouabain to erythrocyte Na,K-ATPase was determined to analyze alterations in this enzyme activity in non-insulin-dependent diabetic patients. A significant (p less than 0.001) reduction of the binding capacity of ouabain was found in erythrocytes obtained from the diabetic patients with polyneuropathy (0.51 +/- 0.02 pmol/10(9) erythrocytes, m +/- SE, n = 14) as compared with the patients without neuropathy (0.67 +/- 0.02, n = 14) or age matched control subjects (0.71 +/- 0.04, n = 11). Accordingly, the effect of an aldose reductase inhibitor (ARI; Ponalrestat) on erythrocyte Na,K-ATPase activity was studied following two or three months oral administration in seven of the diabetic patients with polyneuropathy. After treatment with Ponalrestat the mean binding capacity of ouabain was significantly increased from 0.53 +/- 0.04 to 0.57 +/- 0.03 (p less than 0.05 by paired t-test). Furthermore, enzyme kinetics showed that in normal subjects the apparent Km and Vmax of erythrocyte membrane Na,K-ATPase were 0.51 +/- 0.07 mM (n = 5, m +/- SE) and 7.19 +/- 0.27 nmol Pi/mg protein/min (n = 5, m +/- SE), respectively. The Vmax with 3 mM ATP was significantly (p less than 0.05) decreased in the diabetic patients with polyneuropathy as compared with age-matched control subjects. However, the apparent Km did not change. Finally, the in vitro effect of Ponalrestat was examined in erythrocyte membrane fractions from the diabetic patients with polyneuropathy. The activity of erythrocyte membrane Na,K-ATPase was found to be directly stimulated about 1.2 fold by the addition of pharmacological doses of Ponalrestat (10(-10), 10(-8), 10(-6) M).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561397 TI - [Epidermolysis bullosa and collagenoses]. PMID- 2561398 TI - Effect of lithium on regulation of two molecular forms of Na+,K(+)-ATPase in rat brain. AB - Effects of lithium in vivo and in vitro on the two molecular forms of Na+,K(+) ATPase in rat brain were investigated. Inhibition by strophanthidin, affinity to monovalent cations and cellular localization of the enzyme were used to differentiate the two molecular forms. K+ dependent p-nitrophenylphosphatase activity and strophanthidin inhibition studies revealed selective increase in the activity of low affinity form but not high affinity form of the enzyme following lithium treatment. Na+ sensitivity of neither forms of Na+,K(+)-ATPase was changed but K+ sensitivity of low affinity form was increased due to lithium. Lithium showed biphasic effects on low affinity form of the enzyme; activation at low concentration and inhibition at high concentration. The results suggest that lithium in vivo regulates the concentration of extra cellular potassium by selectively acting at K+ site of low affinity form of the enzyme (astroglial) but not on high affinity form (neuronal enzyme) and leading to changes in neuronal depolarization. PMID- 2561399 TI - Neutrophil myeloperoxidase deficiency associated with canine hepatozoonosis. AB - Hepatozoonosis is a very important disease in dogs in Nigeria. Hepatozoonosis was reported in Nigeria in 18 dogs. The clinical signs included fever, anorexia, loss of weight, lameness, oculonasal discharge and conjunctivitis. Hematologic findings included leukocytosis due to neutrophilia and eosinophilia. Parasitemia varied from 1 to 9% of the circulating neutrophils in the peripheral blood smears of the dogs examined. Hepatozoon canis gametocytes were identified in circulating neutrophils of dogs. Peripheral blood smears from dogs confirmed to have natural H. canis infection were cytochemically stained for myeloperoxidase. Parasitized neutrophils were myeloperoxidase deficient while non-parasitized neutrophils were myeloperoxidase positive. This is considered important, because deficiency of the enzyme may be responsible for poor response of H. canis to chemotherapeutic agents. PMID- 2561400 TI - [Ankyloblepharon filiforme adnatum and associated syndromes]. AB - The authors present 2 cases of ankyloblepharon filiforme adnatum (AFA). In the first case, the ocular anomaly was unaccompagnied by other malformations; in the second patient, there was an associated popliteal pterygium syndrome. The authors review histologic, embryologic features and etiopathogenic theories of AFA and also fully discuss the various syndromes which may be associated with this ocular anomaly. Rosenman's classification is presented. PMID- 2561401 TI - Lymphopenia in primary degenerative dementia. AB - The central nervous and immune systems share a number of common properties, e.g. neurotransmitter receptors. In contrast to age matched healthy and/or hypertensive controls, the total lymphocyte count in 25 carefully diagnosed and staged patients with primary degenerative dementia were significantly reduced and also correlated with the severity of their dementia. Subjects' age, age of dementia onset, or duration of illness were unrelated. Total leukocyte counts were similar between groups. Nutritional status, ACTH, or plasma cortisol in dementia cohort were also not explanatory of their lymphopenia. These results are potentially significant in light of an immune hypothesis for primary degenerative dementia: Alzheimer's type and/or that population's high concomitant risk of infection. PMID- 2561402 TI - Shedding of herpes simplex virus type 1 into saliva after surgery for oral and genital or urological cancer patients. AB - The shedding of herpes simplex virus type-1 (HSV-1) into the saliva was compared in 28 patients with oral cancer and 26 patients with genital or urological cancer. All subjects tested positive for HSV-1 specific antibody. A statistically significant (p less than 0.001) difference was found: infectious viruses were isolated from 12 (39.8%) of the oral cancer patients versus only 2 (7.6%) of the genital or urological patients. This indicates that direct stimulation of peripheral nerves during surgery was responsible for the greater reactivation of HSV-1. PMID- 2561403 TI - The effect of cimetidine on survival of mice infected with herpes simplex virus type 2, murine encephalomyelitis virus and vesicular stomatitis virus infections. AB - The effect of cimetidine on survival was investigated in mice infected with herpes simplex virus type 2 (HSV-2), murine encephalomyelitis virus (GD-VII), and vesicular stomatitis virus (VSV). BALB/c mice, 5 weeks of age, were injected intraperitoneally (i.p.) with 5.5 x 10(5) plaque-forming units (PFU) of virus/0.5 ml, and cimetidine (1 mg/0.5 ml) was administered simultaneously. The survival rates of 80% and 85% in the cimetidine groups were significantly greater than the 10% and 23% for the control groups. The GD-VII- and VSV-infected control mice were dead at 3 days after virus inoculation. However, more cimetidine-treated mice survived than control mice. When anti-mouse T-cell serum or cyclosporine, which is a helper T-cell suppressor, was administered to BALB/c mice; the effect of cimetidine against the HSV-2 infection could be observed. When injected with anti-asialo GM1, BALB/c mice or beige mice with low natural killer (NK) cell activity were not affected by cimetidine. Lastly, cimetidine was shown to activate the cytotoxic action on NK cells. The above results indicate that the antiviral effects of cimetidine depend on NK cell activation. PMID- 2561404 TI - [Pemphigus herpetiformis associated with neoplasm of the lung]. AB - We report a case of a 57 year-old male who developed atypical bullous disease and in whom an underlying carcinoma of the bronchus was found. The cutaneous eruption fulfilled the clinical, histological and immunological features of pemphigus herpetiformis. Reports of pemphigus herpetiformis and internal malignancy are extremely rare. PMID- 2561405 TI - [Paraganglioma (chemodectoma) of the larynx. Presentation of 2 cases]. AB - Two rare cases of laryngeal paraganglioma are reported. The first patient was a 14-year-old girl, that had a new laterocervical paraganglioma, 4 years after the excision of a laryngeal tumor. The other patient was a 33-year-old man. In both the cases, the diagnosis was made on histopathological ground and the tumors displayed the typical histological pattern of paragangliomas: epithelioid tumor cell arranged in nests (Zell-ballen) in a vascular stroma. Neoplastic cells were found to contain argyrophil granules and chromogranin. Laryngeal paragangliomas reported in the literature are 73, they generally are supraglottic, show no sex predilection and prefer the V-VII decades of age. PMID- 2561406 TI - [Extramammary Paget's disease. Immunocytochemical study and histogenetic considerations]. AB - We report a case of extramammary Paget's disease arising in the anogenital region in association with an underlying sweat gland microcarcinoma. Paget's cells were investigated with monoclonal anti-EMA and anti-cytokeratin antigen and with mono- and polyclonal anti-CEA antigen. Positive immunostaining was observed in Paget's cells and in the underlining tumor, whereas keratinocytes and melanocytes did not stain. CEA was also detected in cells and secretions of normal apocrine glands. The immunohistochemical use of polyclonal anti-keratin and anti-S-100 protein antigen is helpful in differentiating Paget's disease from other morphologically similar skin lesions, such as Bowen's disease and superficial spreading melanoma in situ. PMID- 2561407 TI - [Adsorption of dyes as an indicator of the biological activity of mineral dust]. AB - A study is made on the adsorption capacity of several types silicates and quartz dusts, prepared from pure mineral standards with respect to methylene blue and fuchsin. There are differences in the adsorption capacity of the minerals referring to unit surface. The quantity of the adsorption for samples of the same mineral, broken to pieces of different dispersity, also differs. However, between the adsorption capacity of test samples, divided into 3 groups--quartz, zeolite and mixed from the work environment, with respect to methylene blue, and their biological aggressiveness, tested "in vivo" after intratracheal test and the subcutaneous pocket method and the zeolite dusts on diploid culture, an inverse dependence is established. The adsorption capacity of the test dusts from the work environment is many times higher than the examined quartz and zeolite dusts, because of the presence of mineral components in them, especially argillaceous, which have high affinity toward the dye. The inverse dependence established between the adsorption capacity with respect to methylene blue and the biological effect of the tested dusts give grounds to propose the adsorption capacity as one of the induces for studying and hygienic evaluation of different types dusts, which have no components manifesting specific affinity to the dye. PMID- 2561408 TI - Effects of smoking on the synthesis of leukotriene B4 by rat alveolar macrophages. AB - Leukotriene B4 (LTB4) is a potent chemotactic factor for polymorphonuclear leukocytes. It was reported recently that pulmonary macrophages from some mammalian species synthesize and release LTB4. Laviolette et al. reported the decrease of LTB4 synthesis in smokers' alveolar macrophages (AM). In this paper, we report the chronic effects of passive smoking on the production of LTB4 by rat AM. Some rats inhaled tobacco smoke and some inhaled smoke which did not contain nicotine. LTB4 production from AM from these rats was compared. In the tobacco smoking group (n = 12), LTB4 production was 8.0 +/- 2.4 ng/10(6) cells. This value was significantly lower than in the nonsmoking (NS) group (n = 12; 16.5 +/- 4.5 ng/10(6) cells; p less than 0.01) and in the non-nicotinized cigarette smoking (NNCS) group (n = 8; 18.5 +/- 3.5 ng/10(6) cells; p less than 0.01). There were no significant differences between the NS and the NNCS group. These data may suggest that chronic inhalation of nicotine in tobacco smoke decreases LTB4 production activity of rat AM. The precise mechanism of decreased synthesis of LTB4 is not clear but it may have some immunological effects on the lung. PMID- 2561409 TI - Effectiveness of nedocromil sodium versus placebo as additions to routine asthma maintenance therapy: a multicentre, double-blind, group comparative trial. AB - A total of 117 subjects took part in a double-blind, placebo-controlled trial of nedocromil sodium (4 x 4 mg daily by inhalation) in the management of patients with moderately severe bronchial asthma. Treatment was randomly allocated and was continued for 12 weeks following a 2-week baseline period on routine asthma therapy. Statistically significant differences for nedocromil sodium versus placebo were found for diary card asthma severity scores, morning and evening peak flow measurements and daily bronchodilator usage. The clinicians' assessments of asthma severity and global opinions of treatment efficacy were also significantly in favour of nedocromil sodium. Overall, the clinical status of these asthmatic patients was improved by adding nedocromil sodium onto their existing therapy. PMID- 2561410 TI - [Biological evaluation of new hydroxyapatite endodontic cement in vivo. Histopathological and clinico-pathological observation]. AB - Histopathological observations of vital human and monkey dental pulp were performed to evaluate biologically a new root canal filling material APC containing hydroxyapatite. Experimental materials were divided into the following 3 groups. Group 1. Direct application to pulp wound (monkey teeth), 30 and 90 days. Group 2. Direct application to pulp wound (human teeth), 8-477 days. days. Group 3. Root canal filling on pulp stump (human teeth), 7-461 days. After extraction, the experimental teeth were fixed, demineralized, dehydrated and embedded in celloidin, they were prepared as thin sections and submitted to hematoxylin-eosin staining. Light microscopic observations were then made of them. After fixing, some specimens were embedded in resin and prepared in ultra thin sections before being triple stained with tannic acid-ethanol uranium-lead. They were submitted to observation under a HITACHI H-600 transmission electron microscope (TEM). Element analysis was performed on some unstained sections by means of energy dispersive X-ray analysis with a Kevex 7000A. Results 1. Application of APC to monkey dental pulp had no inhibiting effects on the healing progressed, the formation of regeneration hard tissue was observed. APC is believed to be biocompatible with dental pulp tissues. 2. Of the total of 50 cases in which APC was applied to human dental pulp, by the end of the experiment, 42 had occurred no symptoms of any kind. Broked down further, this means 23 (92%) of 25 cases in Group 2 and 19 (76%) of 25 cases in Group 3. Of the 25 cases in Group 2, 24 (96%) were evaluated as clinically good, none (0%) as fair, and 1 (4%) as unsatisfactory. Of the 25 cases in Group 3, 22 (88%) were evaluated as clinically good, 2 (8%) as fair, and 1 (4%) as unsatisfactory. Histopathologically, various pathological findings occurred in both groups of experimental materials. In the early stage, hyperemia, hemorrhage, round-cell infiltration, suppurative inflammation, and resorption of dentin in chamber walls were observed. By the thirtieth postoperative day, pulp cicatrization, formation of new hard tissue, and apposition of dentin on chamber walls were observed. TEM investigation of round-cell infiltration, observed in both group, revealed incursions of hydroxyapatite and bismuth in the macrophages. The tendency for hard tissue newly generated has began on 73 days in Group 2 and on 32 days in Group 3. This finding was made somewhat earlier in Group 2. Thereafter, dental pulp continued in a healthy condition.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2561411 TI - Atrophy of left lobe of the liver associated with hepatolithiasis and cholangiocarcinoma. AB - A case of liver atrophy associated with hepatolithiasis and cholangiocarcinoma is presented. Intraductal calculus and left hepatic lobar atrophy were seen on both ultrasound and computerised tomography examinations. The lobar atrophy was accompanied by hypoplasia of left common hepatic artery. Cholangiocarcinoma was diagnosed on histology. There have been several reported cases of hepatolithiasis and cholangiocarcinoma. To our knowledge, there has been only one reported case of hepatolithiasis and cholangiocarcinoma with liver atrophy (Nishihara K et al 1986) but this was not angiographically demonstrated. PMID- 2561412 TI - [The cobalt lung in diamond cutters: a new disease]. AB - Although for forty years already broncho-pulmonary pathology has been described in workers exposed to hard-metal (i.e. alloys of tungsten carbide and cobalt) and although cobalt is considered the offending agent of this hazard, these abnormalities have almost not been found after exposure to cobalt alone except in animal experiments. Recently we detected clearcut broncho-pulmonary pathology in 48 diamond polishers (i.e. nearly 1% of those exposed) attributable to the ultrafine cobalt dust from the cutting surface of polishing disks, in which it was used as a cementing matrix for microdiamonds without any tungsten carbide. Nineteen of these patients presented with a fibrosing alveolitis documented in 6 by lung biopsy and in 12 by broncho-alveolar lavage, both of which revealed characteristic multinucleated giant cells. Thirteen suffered from asthma of occupational origin, in 9 proven by cobalt-inhalation tests, and in 5 by peak flow measurements at the workplace. Sixteen had mixed bronchial and alveolar pathology or were incompletely documented. A cross-sectional study in about 200 diamond polishers showed a significant correlation between exposure to cobalt and decrease in lung function. The strikingly harmful effects of cobalt can be explained by the chronic exposure to very small particles with markedly increased solubility. The pathogenesis of the broncho-pulmonary pathology may be attributed to the cytotoxic as well as to the sensitising (i.e. allergic and/or idiosyncratic) actions of cobalt. PMID- 2561413 TI - [Deficiency in hepatic uptake of glucose in chronic diabetes mellitus]. AB - In insulin-dependent diabetes mellitus there is a deficient post-prandial uptake of glucose and storage as glycogen in the liver. This impairment is due to an intrinsic hepatic defect that has been investigated with the use of isolated liver cells. Glycogen synthase catalyzes the rate-limiting step in the synthesis of glycogen. In response to an increased glucose concentration, this enzyme is activated in normal hepatocytes through dephosphorylation of seryl residues by a glycogen-bound "protein phosphatase G". Hepatocytes isolated from alloxan diabetes rats have lost the ability to activate glycogen synthase in response to an increased glucose concentration. The magnitude of the latter defect corresponds to the severity of the diabetes, as judged from the level of glycaemia. The defect is explained by an impaired function of protein phosphatase G. The latter enzyme consists of a catalytic subunit (37 kDa) associated with a large glycogen-binding subunit (161 kDa) and other regulatory polypeptides. It appears that in diabetes an essential regulatory subunit is deficient. Studies in animals with distinct types of spontaneous diabetes revealed that lack of insulin, rather than chronic hyperglycaemia, explains the deficient activity of protein phosphatase G. PMID- 2561414 TI - [Herpetic manifestations: diagnosis, treatment, prevention]. AB - Herpes virus infection is an extremely common disease that affects between 90 to 100% of the population above the age of 15. The authors propose studying successively the clinical and virological diagnosis of this infection, it's treatment, and finally it's prevention in the dental clinic where there is a large possibility of transmitting the infective diseases. PMID- 2561415 TI - A study of 5'-nucleotidase activity in subcellular fractions of rat cerebellum after the administration of the convulsant 3-mercaptopropionic acid. AB - The activity of 5'-nucleotidase in cerebellum subcellular fractions after the administration of the convulsant 3-mercaptopropionic acid was studied. This membrane enzyme presented an increase in its activity in certain fractions containing nerve endings and microsomes (Mic20, Mic100) in seizure and postseizure periods. 5'-nucleotidase activity in nuclear and myelin fractions did not show differences between the control and treated fractions. On the other hand, a decreased activity in the crude mitochondrial fraction and in a nuclear subfraction was found. It is suggested that the changes in the enzyme activity in some cerebellum fractions might be related to structural alterations previously observed in this laboratory and with the anticonvulsant actions of adenosine. PMID- 2561416 TI - Experimental subarachnoid hemorrhage. Lipid peroxidation and Na+,K(+)-ATPase in different rat brain areas. AB - Subarachnoid hemorrhage (SAH) was produced in Sprague Dawley rats by injection of 0.30 mL of autologous arterial blood into the cisterna magna. Tissue lipid peroxide, quantified as thiobarbituric acid reactive material (TBAR), and Na+,K(+)-ATPase activity were assayed in three different rat brain areas (cerebral cortex, hippocampus, and brain stem) of sham-operated rats and in four hemorrhagic rat groups at 30 min, 1 h, 6 h, and 2 d after SAH. Na+,K(+)-ATPase activity decreased in the cerebral cortex at 30 min, 1 h, and 6 h and in the brain stem at 1 h after SAH induction, whereas enzymatic activity was unchanged in the hippocampus. There was no evident difference in lipid peroxide content between sham-operated animals and hemorrhagic animals. These results indicate that little modifications in lipid peroxidative process (as expressed in TBAR) are not responsible for changes in the ATPase activity. PMID- 2561417 TI - Myxoid chondrosarcoma of the mediastinum. AB - We report a unique case of primary myxoid chondrosarcoma in the posterior mediastinum. This lesion clinically mimicked a neurogenic tumor due to its location and dumbbell appearance. The histogenesis of this tumor is discussed and a skeletal origin from the thoracic vertebrae is suggested. It is concluded that myxoid chondrosarcoma has to be distinguished for the differential diagnosis among the long list of myxoid tumors arising in the posterior mediastinum. PMID- 2561418 TI - [Results of a phase II study of the combination of cis-platinum and 5-FU in epidermoid or undifferentiated large cell bronchial cancer]. AB - The aim of this study was to test cisplatinum and 5-FU chemotherapy. Thirty-five patients with epidermoid or undifferentiated lung carcinoma were entered in a multicentric phase II trial, in an attempt to further define the activity and toxicity of this association. None of them had been previously treated by chemotherapy. The dosage schedule was cisplatinum 100 mg/m2 D1 and 5-FU 1 g/m2 D1 to D5 every 3 weeks for 3 courses before evaluation. There were CR: 2, PR: 10, for a total response rate of 35%. Median survival was 7 months. Tolerance was acceptable. We conclude that this association can be safely administered, but that the results are not superior to others previously reported. Further studies are required to define the activity of a cisplatinum, 5-FU and radiotherapy association. PMID- 2561419 TI - [Cloning and gene expression determining phenol breakdown in Pseudomonas putida strains]. PMID- 2561420 TI - [The effect of alpha-beta-11-hydroxysaxitoxin sulfate on the sodium currents of mammalian neurons]. PMID- 2561421 TI - Didemnin B. Conformation and dynamics of an antitumour and antiviral depsipeptide studied in solution by 1H and 13C. n.m.r. spectroscopy. AB - The solution conformation of didemnin B, the most potent member of a family of depsipeptides that shows antitumour, antiviral, and immunosuppressive activity, has been studied in chloroform solution using n.m.r. spectroscopy. 1H and 13C spectra have been assigned from analysis of a number of two-dimensional homonuclear and heteronuclear chemical shift correlation experiments which confirm the recently corrected primary structure of the molecule. The conformation of the peptide has been deduced from measurements of the temperature dependence of the NH chemical shifts, analysis of coupling constant data and primarily through NOE effects observed in the rotating frame. Interproton distance bounds determined from a quantitative analysis of the ROE data provide 41 constraints from which a family of closely related structures were calculated using distance geometry algorithms. A type II beta-turn involving residues Thr6, Leu7, and Pro8 is well represented in the computed conformers as is a hydrogen bonding interaction between the NH of Leu3 and the carbonyl oxygen of Thr6. This latter interaction causes the linear portion of the structure to fold back over the depsipeptide ring, imparting to it a degree of structural stability as well as giving the molecule a somewhat globular character. Only one transannular hydrogen bond, between Ist1 NH and Leu3 carbonyl, stabilizes the conformation of the depsipeptide, which has an irregular non-planar configuration. The small temperature coefficients (less than 2.0 x 10(-3) ppm/degrees C) for the NHs of Ist1 and Leu3 are consistent with their involvement in these hydrogen bonding interactions. We find that many of the structural features observed in the crystalline form of didemnin B are conserved in solution. Analysis of the 13C spin-lattice relaxation rates of the protonated carbons reveals small variations in effective correlation times at specific sites in the molecule. The data suggests that the peptide segment encompassing residues Leu3 through to Thr6 is in a more motionally restricted part of the structure. PMID- 2561422 TI - Further characterization of a size control gene in Saccharomyces cerevisiae. AB - The DAF1-1 mutation reduces cell size and reduces or eliminates G1 phase in Saccharomyces cerevisiae, and results in alpha-factor resistance. DAF1-1 cells transferred into low cycloheximide express an increased G1 phase in their cycle, suggesting that G1 regulation is present but cryptic in the DAF1-1 cycle in rich medium. DAF1-1 reduces cell size by the criterion of RNA content per cell as well as cell volume. The alpha-factor resistance of DAF1-1 cannot be suppressed by bypassing the pheromone-receptor interaction with 'signalling-constitutive' mutations, suggesting that pheromone binding and initial signalling is normal in DAF1-1 strains, but that division arrest in response to the signal is specifically defective. Consistent with this idea, the cdc28-13 mutation significantly suppresses DAF1-1 alpha-factor resistance at permissive temperature; CDC28 is a gene required specifically for START and the G1/S transition, and does not affect pheromone response. Genetic results additional to those previously reported confirm that the wild-type dafl+/WHI1 gene is non essential; this result may be surprising since the gene product is apparently rate-limiting for the G1/S transition: its deletion increases cell size, and multiple copies decrease cell size. PMID- 2561423 TI - Cellular and viral control of the initiation of DNA replication. AB - Cell-free replication of SV40 DNA in extracts prepared from S phase cells is at least 20-fold more efficient than in extracts from G1 cells. The increased activity of S phase extracts correlates with the presence of an S phase-specific cellular factor that enhances DNA unwinding at the replication origin. This change in origin-DNA structure during the initiation of SV40 replication proceeds through at least three discrete steps which can be distinguished by their extent of topologic unwinding (linking differences of -1, -2 and -5). Specific DNA elements flanking the core origin enhance replication in vivo and facilitate the formation of the pre-initiation complexes, indicating that formation of these underwound conformations may be the limiting step in the initiation of DNA synthesis. In addition, the factor that activates DNA replication in extracts from S phase cells also enhances the formation of the most highly underwound -5 pre-initiation complex. These observations suggest that during SV40 replication, formation of the rate-limiting pre-initiation complex is the focus of at least three regulatory elements. Two of these are DNA sequences flanking the replication origin and the third is a cellular factor specific to the S phase cell. PMID- 2561424 TI - Gene products required for chromosome separation. AB - Gene products required for mitotic chromosome separation in the fission yeast Schizosaccharomyces pombe are described. They have been identified by two distinct strategies of mutant isolation, followed by gene cloning and immunochemical characterization of gene products. The roles of four representative genes, namely nda3+, nuc2+, top2+ and dis2+, encoding beta tubulin, a nuclear scaffold-like protein, DNA topoisomerase II and type-1 protein phosphatase, respectively, are discussed in regard to the mechanisms and control of chromosome separation. PMID- 2561425 TI - Cis- and trans-acting factors affecting the structure of yeast centromeres. AB - The isolation of yeast centromeres has provided the opportunity to describe the molecular structure of chromosome attachments to the mitotic spindle. Nucleolytic probes of chromatin structure and analysis of genetically altered centromeres have been used to determine the primary sequence and positional requirements for assembly of this chromosomal domain into a functional unit. Structural analysis of the centromere in wild-type and mutant strains provides the first indication of specific proteins associated with centromere DNA in vivo. PMID- 2561426 TI - M phase-promoting factor: its identification as the M phase-specific H1 histone kinase and its activation by dephosphorylation. AB - A major protein kinase independent of Ca2+, cyclic nucleotide or diacylglycerol, the activity of which becomes maximal when cells enter M phase, decreases at ana telophase, and is low during interphase, has been purified to near homogeneity from starfish oocytes and its catalytic subunit identified as p34cdc2. M phase promoting factor (MPF) was found to co-purify with the M phase-specific kinase throughout its purification. p34cdc2 does not have to be associated with any specific protein for expression of H1 histone kinase or MPF activities. When p34cdc2 is phosphorylated its protein kinase activity is inhibited, preventing entry into M phase, but once p34cdc2 becomes dephosphorylated, its protein kinase activity increases and M phase is initiated. A second peak of MPF activity was separated from p34cdc2 in the ammonium sulfate fraction treated with ATP-gamma-S. It induced p34cdc2 dephosphorylation and the concomitant stimulation of its kinase activity when injected in Xenopus or starfish oocytes. PMID- 2561427 TI - Temporal regulation of cdc2 mitotic kinase activity and cyclin degradation in cell-free extracts of Xenopus eggs. AB - In cleaving Xenopus eggs, the cell division cycle is abbreviated to a rapid succession of S and M phases. During mitosis a number of proteins show increased phosphorylation due to the activation of a histone H1 kinase, the homologue of the cdc2+ gene product of the yeast Schizosaccharomyces pombe. We have studied the regulation of the activity of this enzyme in cell-free extracts of Xenopus eggs. In extracts of activated eggs incubated at 22 degrees C, histone H1 kinase activity shows two peaks of activation and disappearance. Activation occurs in two stages. The first stage requires protein synthesis, whereas the second does not. The second stage of activation involves post-translational activation of the kinase. Kinase activity rises to a peak and then abruptly disappears. Added sea urchin cyclin is degraded at the time of disappearance of kinase activity. The oscillation in kinase activity is then repeated, usually with lower amplitude. Post-translational activation of the kinase requires a membrane-containing particulate cellular component, whose role has yet to be defined. The kinase can still be activated in the presence of EDTA or in the presence of the ATP analogue, 6-dimethylaminopurine, which implies that phosphorylation of the kinase complex is not required for activation. Under these conditions, however, the kinase activity does not show its normal sudden disappearance, and added cyclin is perfectly stable. These observations are consistent with the idea that post translational activation of the kinase involves protein phosphatase activity, whereas switching off the kinase requires an ATP-Mg2(+)-dependent reaction, perhaps due to protein phosphorylation PMID- 2561428 TI - Indeloxazine augments long-term potentiation in the mossy fiber-CA3 system of guinea pig hippocampal slices. AB - The effects of indeloxazine and piracetam, which reportedly prevent experimental impairments of memory and learning, on long-term potentiation (LTP) of the population spike in the mossy fiber-CA3 system were studied in vitro using the guinea pig hippocampal slices. Indeloxazine (10(-6) M) or piracetam (10(-4) M) significantly augmented the LTP and the effect of indeloxazine on the LTP was approximately 100 times as potent as that of piracetam. PMID- 2561429 TI - [Rapid diagnosis of cytomegalovirus in immunocompromised patients using monoclonal antibodies that detect early viral antigens]. AB - A technique was applied to detect early fluorescent antigens (DEFA) of cytomegalovirus (CMV) using the E13 monoclonal antibodies in 52 immuno compromised patients hospitalized in the Nephrology Institute of Havana. Of the 75 urine or blood (buffy coat) samples taken, 15 were found positive to CMV. Using classical diploid human fibroblast isolation technique, 12 CMV strains were isolation of previously detected positive samples by DEFA. In addition, CMV was isolated from one sample reported to be negative by DEFA. A coincidence of 80% was found between both techniques. With the ELISA test, all the sample studied have IgG antibodies to CMV. PMID- 2561430 TI - The blood flow in the periodontal ligament regulated by the sympathetic and sensory nerves in the cat. AB - This study was carried out to investigate the nervous control of the blood flow in the periodontal ligament measured by laser Doppler flowmeter. Ten adult cats were anesthetized with pentobarbital sodium (initial dose of 30 mg/kg, i.v. and maintenance dose of 5 mg/kg, i.v.). After enucleating the left eye ball, the superior alveolar nerve was exposed. The bone overlying the labial aspect of the left maxillary canine tooth root was pared away until a transparent layer of bone was left covering the periodontal ligament. A laser light from a probe of the flowmeter fixed at the tooth was beamed through the thinned bone. Three different patterns of responses were observed following the electrical stimulation of the distal end of the cut superior alveolar nerve: an increasing, a decreasing and a biphasic change of blood flow. The application of capsaicin onto the superior alveolar nerve reduced the response of blood flow increase but had no effect on the response of blood flow decrease. On the other hand, the response of blood flow decrease was completely inhibited by the pretreatment with phentolamine while the response of blood flow increase was not affected. The present results suggest that blood flow in the periodontal ligament of cats is controlled by sympathetic alpha-adrenergic fibers for vasoconstriction and by sensory fibers for vasodilation. PMID- 2561431 TI - Calcitonin gene-related peptide (CGRP) in the regulation of salivary secretion. AB - The occurrence of calcitonin gene-related peptide (CGRP) in nerves in the submandibular gland was studied by using indirect immunofluorescence technique. The functional significance of CGRP was further evaluated by using an in vitro method for demonstration of protein and peroxidase secretion. CGRP-immunoreactive nerve fibers were observed mainly around large ducts and around blood vessels in the stromal part of submandibular gland. A moderate amount of CGRP immunoreactivity were seen around the secretory acini as well. In vitro, CGRP increased significantly total protein and peroxidase secretion. SDS gel electrophoresis revealed 20 different protein bands both in normal and stimulated samples. After CGRP-stimulation 13 out of these 20 bands showed more intensive staining in this semi-quantitative analysis. The present results demonstrate that the secretory elements of the rat submandibular gland are innervated by CGRP-IR nerve fibers and that CGRP has a significant effect on protein secretion in vitro. Thus our results support the view that CGRP has a direct stimulatory effect on salivary secretion. PMID- 2561432 TI - [Fibrolamellar hepatocellular carcinoma: a clinical report with paraneoplastic hyperthyroidism (apropos of a case)]. AB - The case of a 17 years old female patient with fibrolamellar hepatocarcinoma and several paraneoplastic signs is reported. The clinical picture was hyperthyroidism like; subsequently metastasis lesions in lungs and muscular tissue (gluteal region) developed. Hyperthyroidism as clinical picture has been reported only once before us for the available literature reveals. PMID- 2561433 TI - [Measurement of angiotensin I convertase activity in the serum of the rat]. AB - A fluorimetric method has been developed for the determination of activity of angiotensin I convertase in rat blood serum. The method proved highly sensitive. It enables determination of the activity of angiotensin I converting enzyme in 0.01 cm3 of blood serum. The method may be applied to in vitro and in vivo testing of inhibitors of angiotensin I convertase. PMID- 2561434 TI - Influence of fat and feed level on fiber digestibility in vitro and in sacco and on volatile fatty acid proportions in the rumen. PMID- 2561435 TI - Comparative aspects of fibre fermentation in mammals. PMID- 2561436 TI - Influence of the physical form of diet on chewing activity and reticulo-rumen motility in cows. PMID- 2561437 TI - Determination of gastrointestinal passage of fluid and fibre in the rabbit using single-dose nonabsorbable markers. PMID- 2561438 TI - Development of the porcine gastric proteases. The effect of age, ACTH-treatment and early weaning. PMID- 2561439 TI - [Value of the CRF test in primary and secondary adrenal insufficiency]. AB - The ACTH and cortisol responses after CRF administration were studied in 21 patients with adrenal failure and compared with those obtained in 15 control subjects. In patients with Addison's disease, cortisol and ACTH responses were respectively abolished or excessive. In patients with secondary adrenal failure, a unique bolus injection of CRF allows the differentiation between pituitary or hypothalamic secretory defect. With regard to previous reports, the usefulness of CRF test in primary and secondary adrenal failure is stressed. PMID- 2561440 TI - Chelation of lead during co-exposure to ethanol. AB - Efficacy of calcium disodium EDTA, D-penicillamine (DPA), 2,3 dimercaptosuccinic acid (DMSA), and alpha-mercapto-beta-(2-furyl) acrylic acid (MFA) to reduce the body burden of lead and restore the altered biochemical variables in lead or lead + ethanol administered rats was investigated. The investigation was aimed to suggest suitable prophylaxis of lead intoxication prevalent among workers co exposed to lead and alcohol ingestion. Administration of lead (10 mg/kg, oral, once daily for 8 weeks) produced a significant inhibition in the activity of blood delta-aminolevulinic acid dehydratase (ALAD), elevation in the blood zinc protoporphyrin (ZPP) and urinary elimination of lead and delta-aminolevulinic acid (ALA). Lead contents of blood, liver, kidney and brain were also significantly higher than the normal control. The above changes were more marked in animals co-exposed to lead + ethanol (20% in drinking water) compared to lead alone. All the chelators were effective in increasing the urinary lead elimination, reducing the above biochemical alterations and lead contents of tissues. The order of effectiveness being DMSA greater than Calcium disodium EDTA greater than DPA greater than MFA. However, the protection was more noticeable in animals treated with lead alone than with lead and ethanol. PMID- 2561441 TI - Reduction of cyclic AMP phosphodiesterase activity of several subcellular fractions of rat brain after pretreatment with phospholipase C. AB - Cyclic AMP phosphodiesterase (PDE) activity was assayed in the plasma membrane, mitochondrial and microsomal fractions of rat brain. The specific activity of the enzyme was highest in the plasma membrane fraction followed by mitochondrial and then the microsomal fraction. Phosphodiesterase activity of all three fractions was reduced after pretreatment with lecithinase C (PCase) from Clostridium perfringens but less markedly affected by the pretreatment with sphingomyelinase (SMase) from human placenta. The PDE activity of the plasma membrane fraction was more sensitive to PCase treatment compared with the other two particulate fractions, which showed only a slight loss of activity. Temperature seemed to affect PDE activity of the plasma membrane. The enzyme was quite stable at 30 degrees C but its activity dropped by approximately 46% at 37 degrees C after 90 min of incubation. Pretreatment of the plasma membrane at 30 degrees C with PCase at a concentration of more than 5 U caused a marked loss of PDE activity and the decrease in activity reached a plateau at concentrations above 10 U. PMID- 2561442 TI - Evidence that coupling factor B is bound to the matrix side of the inner mitochondrial membrane. AB - Rotenone-sensitive NADH dehydrogenase activity and Lubrol stimulation of cytochrome oxidase activity were measured to assess the opposite membrane polarity of beef heart mitoplast and inside-out particle preparations. The ATP-Pi exchange activity of mitoplasts was not affected by their incubation at pH 8.9 in the presence of 5 mM EDTA (a treatment known to extract coupling factor B (F beta) from submitochondrial particles), nor was it stimulated by the addition of F beta to intact and alkaline treated mitoplast preparations. In contrast, the exchange activity of inside-out particles was decreased 18 fold by the alkaline/EDTA treatment and was almost completely restored by the addition of F beta to F beta-depleted particles. From these results it is concluded that in beef heart mitochondria, the coupling factor F beta is bound to the matrix-side of the inner mitochondrial membrane. PMID- 2561443 TI - Superoxide--driven oxidation of quercetin and a simple sensitive assay for determination of superoxide dismutase. AB - Oxidation of quercetin at pH 10 was shown to be a free radical chain reaction involving superoxide and hence inhibitable by superoxide dismutase (SOD) (EC 1.15.1.1). The degree of inhibition of quercetin oxidation was a function of SOD concentration, and fifty percent inhibition was produced by approximately 1.5 ng/ml of pure enzyme. This reaction proved to be a very useful tool for a rapid and highly sensitive measurement of SOD in crude tissue extracts and other biological samples. PMID- 2561444 TI - Comparison of the inhibitions of proliferation of normal and psoriatic fibroblasts by 1 alpha,25-dihydroxyvitamin D3 and synthetic analogues of vitamin D3 with an oxygen atom in their side chain. AB - The inhibitory effects of 1 alpha,25-(OH)2D3 and synthetic oxa-derivatives of vitamin D3 on growth of normal and psoriatic fibroblasts in culture were compared. Proliferation of normal fibroblasts was strongly inhibited by these new compounds in the following order: 22-oxa-1 alpha,25-(OH)2D3 greater than 22-oxa-1 alpha-(OH)D3 greater than 1 alpha,25-(OH)2D3 greater than 20-oxa-1 alpha,25 (OH)2D3. 22-Oxa-1 alpha,25-(OH)2D3 was about 10-times more inhibitory than 1 alpha,25-(OH)2D3. Proliferation of psoriatic fibroblasts was not inhibited by 1 alpha,25-(OH)2D3 at concentrations of up to 10(-6) M, but was suppressed by 10( 8)-10(-6) M 22-oxa-1 alpha,25-(OH)2D3 and 10(-6) M 22-oxa-1 alpha-(OH)D3. These results suggest that oxa-derivatives of vitamin D3, especially 22-oxa-1 alpha,25 (OH)2D3, should be useful in further studies on the cause and treatment of psoriasis. PMID- 2561445 TI - Yeast genomic clones encoding polypeptides immunologically related to an 18 kDa subunit of mitochondrial ATP synthase. AB - A yeast genomic library in the bacteriophage expression vector lambda gt11 was screened with a polyclonal anti-holo-ATPase antiserum resulting in the isolation of 54 immunoreactive clones. Four of these phage clones express in bacteria a polypeptide antigenically related to an 18 kDa subunit (P18) of the yeast mitochondrial ATPase complex. Molecular analysis of the yeast DNA inserts in these phage clones revealed two classes of yeast DNA that share little homology at the nucleotide sequence level and therefore may represent distinct separate genes. The polypeptides potentially encoded by these yeast DNA segments do show scattered short blocks of strong amino acid sequence homology, which may underlie the observed immunochemical relatedness between the proteins expressed in bacteria. PMID- 2561446 TI - Interaction of S-activated enkephalin analogs with opiate receptors. AB - Enkephalin analogs containing a thiol activated by a thiomethyl (SCH3)*** or 3 nitro-2-pyridinesulfenyl (Npys) group were synthesized. Incubation of such S activated enkephalin analogs as [D-Ala2, Leu(CH2S)SCH(3)5]enkephalin or [D Ala2,Leu(CH2S)Npys5]enkephalin with guinea pig ileum (GPI) resulted in the continuous stimulation of the mu opiate receptors. This sustained GPI-activity was completely reversed with the antagonist naloxone, while subsequent washings elicited again the full enkephalin activity. When GPI showing full enkephalin activity was incubated with 1 mM dithiothreitol, about 70% of the activity was eliminated. Examination of enkephalin analogs containing Cys(Npys) at position 1, 5, or 6 suggested that no other thiols occur near the enkephalin binding site of the mu receptor. From these results, it is considered that only one thiol group exists near the binding site of the mu receptor in GPI. Similar results were also obtained for the mu receptors in mouse vas deferens. PMID- 2561447 TI - Forskolin stimulation of pepsinogen secretion from frog esophageal mucosa is partly mediated by intrinsic cholinergic neurons. AB - Forskolin was found to stimulate pepsinogen secretion from frog esophageal mucosa. The stimulation was dose-dependent and was accompanied with a great increase in tissue cAMP content. The response to forskolin mimicked the action of bethanechol and was not additive with bethanechol. The stimulatory effect of forskolin was inhibited by 50% in the presence of either atropine or tetrodotoxin. On the other hand, incubation in a calcium-free medium not only reduced the response to forskolin by 45% but also eliminated the influence of atropine and tetrodotoxin. These results indicate that forskolin may stimulate pepsinogen secretion from the frog esophageal mucosa via activating adenylate cyclase, and part of its effect may arise from eliciting acetylcholine release from the intrinsic neurons. PMID- 2561448 TI - Thialysine utilization for protein synthesis by an exponentially growing E. coli culture. AB - The extent of protein lysine substitution by thialysine in E. coli cells grown in media containing the analog depends on the time interval the cells are grown in the presence of analog and on the analog concentration in the medium. By calculating the percent of lysine substitution in newly synthesized proteins it was shown that this reaches, after one cell doubling in the presence of analog, a maximum which is 17% in the cells grown with 0.1 or 0.2 mM thialysine and 8% in cells grown with 0.05 mM thialysine. Proteins synthesized in the presence of analog in the concentration range 0.05-0.2 mM show similar stability to those synthesized in the absence of analog. The extent of analog incorporation into newly synthesized proteins, as regards both the time course and the dependence on analog concentration in the medium, is strictly related to the extent of the repression of AK III, the first enzyme of lysine biosynthetic pathway. PMID- 2561449 TI - A comparative kinetic study of bovine calmodulin-dependent cyclic nucleotide phosphodiesterase isozymes utilizing cAMP, cGMP and their 2'-O-anthraniloyl-,2'-O (N-methylanthraniloyl)-derivatives as substrates. AB - Calmodulin-dependent 3',5'-cyclic nucleotide phosphodiesterase (EC 3.1.4.17) Mr 63,000 and Mr 60,000 from the brain as well as Mr approximately 59,000 species from the heart, have been compared with respect to their steady-state kinetic parameters for the hydrolysis of cAMP, cGMP and their 2'-O-anthraniloyl- and 2'-O (N-methylanthraniloyl)-derivatives. Kinetic studies with the native substrates indicate high Mr brain enzyme to be cGMP specific whereas low Mr brain and heart enzymes to be nonspecific. In addition, the isozymes studied here appear to be kinetically distinct from those previously isolated form bovine brain tissues. Substitution at 2'-O-position of the cyclic nucleotides gave rise to Vmax values ranging 1-11% of those observed with the native substrates, with minimal effect on Km. The isozymes with exception of heart isoform gave higher Km and Vmax with the anthraniloyl derivatives. This effect is thought to be related to the formation of an intramolecular hydrogen bond which leads to decreased electrostatic interactions between the active-site side chains and the pseudo substrates. PMID- 2561450 TI - Sulfation of a gamma-chain variant of human fibrinogen. AB - Biosynthetic sulfation of human fibrinogen was investigated using a hepatoma derived cell line in culture. Very little [35]sulfate was incorporated into the major forms of the A alpha, B beta, or gamma-chains of fibrinogen, but there was a labeled peptide chain with electrophoretic mobility intermediate between the B beta and gamma-chains. Base hydrolysis of the sulfate-labeled product released tyrosine sulfate. The labeled peptide is identified as a form of gamma-chain by its resistance to proteolysis during extended periods of incubation, in contrast with A alpha and B beta-chains which are converted to smaller forms. The results indicate that human fibrinogen contains tyrosine sulfate primarily within a variant form of the gamma-chain. PMID- 2561451 TI - Site specific antibodies directed to the ATP binding region of some kinases. AB - Polyclonal antibodies raised in rabbits to ATP-requiring enzymes such as 3 phosphoglycerate kinase show cross-reactivity against other unrelated kinases. Our results show that rabbit polyclonal antiserum possesses antibodies that recognize an antigenic site at the ATP binding region of kinases. A classical immunotitration curve was obtained when hexokinase was titrated against anti myokinase IgG. The immunoinhibitions was reversed in the presence of small concentration of ATP. This cross-reactivity between site specific antibody and unrelated kinase demonstrates the existence of an antigenic site around the ATP binding region. Our proposal of the existence of a common antigenic determinant in the ATP binding region is in agreement with the finding of a common structural domain that binds ATP. PMID- 2561452 TI - Determination genes. PMID- 2561453 TI - Transcription control and differentiation: the HLH family, c-myc and C/EBP. PMID- 2561454 TI - Extracellular signals and intracellular transduction pathways regulating Dictyostelium development. PMID- 2561455 TI - Protein modification: phosphorylation on tyrosine residues. PMID- 2561456 TI - Regulation of protein degradation rates in eukaryotes. PMID- 2561457 TI - Surgical treatment of lung carcinoma involving the chest wall. AB - From 1969 to 1986, 97 patients with chest wall invasion by lung carcinoma (excluding superior sulcus tumours) underwent surgical resection in two hospitals, La Paz (Madrid) and La Fe (Valencia). The same surgical policy was used in both thoracic surgical units: extrapleural pulmonary resection when tumour involved only the parietal pleura (N = 36), and en bloc chest wall resection when the carcinoma extended into the ribs and intercostal muscles (N = 61). The tumour histology was classified according the WHO criteria. Lobectomy or bilobectomy was carried out in 72%, pneumonectomy in 18% and segmentectomy or wedge resection in 10% of the patients. The perioperative mortality was higher in the en bloc resection group 9/61 (15%) versus 2/36 (6%) for extrapleural dissection. The node staging was NO in 58/97 (60%), N1 in 16/97 (16%) and N2 in 23/97 (24%). The probability of survival was calculated by the Kaplan-Meier method collecting data from the perioperative survivors only. The overall 5-year survival was 23% with no significant differences between the en bloc resection and the extrapleural lung resection groups. The most important predictor of survival was the node stage. The 5-year survival for N1 and N2 were 8% and 6%, respectively. These percentages increased to 34% when N0 patients were considered. Other predictors of survival were not significant. The authors conclude that either extrapleural or en bloc chest wall resection are both valid procedures which may be used depending on the depth of local invasion. PMID- 2561458 TI - Excitatory amino acids--quo vadis? PMID- 2561459 TI - PAF-acether, superoxide anion and beta-glucuronidase as parameters of polymorphonuclear cell activation associated with cardiac surgery and cardiopulmonary bypass. AB - 1. Seven patients submitted to myocardial revascularization surgery with cardiopulmonary bypass were studied. Blood samples were obtained immediately before and 24 h after surgery. The parameters studied were the production of platelet activating factor (PAF-acether) and superoxide anion, cellular beta glucuronidase activity as well as polymorphonuclear cell (PMN) and platelet counts. 2. Twenty-four h after surgery, there was a 54% decrease in platelet number (P less than 0.005), a 121% increase in PMN number (P less than 0.005), a 353% increase in PAF-acether (P less than 0.01), a 211% increase in superoxide anion (O2-) and a 104% increase in beta-glucuronidase (P less than 0.05) levels when compared with the pre-surgery levels. 3. The present results indicate that PMN are more reactive after surgery with cardiopulmonary bypass. PMID- 2561460 TI - Characterization of the receptors responsible for the diphasic effect of bradykinin in rat duodenum. AB - The response of the rat duodenum to bradykinin (BK) consists of relaxant and contractile components, which have been attributed to different receptor types. To characterize the receptors responsible for this diphasic response we studied the effects of BK analogues known to act on B1 or B2 receptors in other systems. DesArg9-Leu8-BK and Thi5,8DPhe7-BK presented only relaxant and only contractile effects, respectively, whereas DArg0Hyp3Thi5,8DPhe7-BK was a potent antagonist of the relaxation (but not of the contraction) induced by BK. Our results show that the relaxant and contractile components of the rat duodenum's response to BK are due to B2 and B1 receptor subtypes, respectively. PMID- 2561461 TI - Characteristics of toad bladder urinary acidification. AB - 1. The urinary acification system present in the bladder of the toad Bufo marinus ictericus was investigated by means of an improved technique of mounting the membrane that minimizes the edge damage when the bladder is placed as a sheet between lucite half chambers. 2. In ouabain-treated bladders in the absence of exogenous CO2, acidification rates were similar to those observed for turtle bladder. 3. The presence of 1% CO2 at the serosal gas phase increased proton secretion (JH). Stimulation of JH was also observed when the cell pH was decreased by back diffusion of salicylic acid added to the mucosal (M) compartment. 4. The estimate of the passive proton permeability of this epithelium as a whole yielded values around 1 x 10(-4) cm/s. The maximum pH gradient that could be established across the membrane in the short circuit condition (about 3 pH units) was taken as the apparent proton-motive force (PMF') of the system and these values were similar to those observed in the turtle bladder. 5. Luminal membrane depolarization caused by substitution of NaCl by KCl in Ringer solution led to an increase in JH at M pH = 7.3 without altering the PMF', which suggests that the electrical potential difference across the luminal membrane in ouabain-treated bladders is negligible when M pH is sufficiently acid to abolish H+ secretion. PMID- 2561462 TI - Synthesis and secretion of plasminogen activators and collagenases in human cells transformed by Kirsten murine sarcoma virus and N-methyl-N'-nitro-N nitrosoguanidine. AB - The secretion of elevated levels of proteinases is considered to be a distinct property of most transformed cells. The cellular and secreted levels of plasminogen activators and collagenases have been examined in the nonmalignant human osteosarcoma (HOS), the malignant Kirsten murine sarcoma virus transformed (KHOS/NP), the temperature sensitive revertant of virus transformed HOS (KHOS 240S) and N-methyl-N'-nitro-N-nitrosoguanidine transformed HOS (MNNG/HOS) clones. Virus and MNNG transformed clones exhibit 100- and 7-fold higher cellular and and 270- and 30-fold higher extracellular plasminogen activator (PA) activity as compared with untransformed HOS controls. The cellular PA activity of the revertant clone is similar to but the secreted level is slightly higher than the HOS controls. SDS-PAGE in the presence of casein and plasminogen is consistent with the major PA species of urinary type (u-PA) and with the absence of PA inhibitor in the parent and revertant clones. The cellular levels of active collagenase are low in all the clones. However, on activation by trypsin, the two active collagenase bands of similar intensity are observed for all the lines in SDS-PAGE in the presence of gelatin. While there appears to be some elevation of secreted collagenase prior to trypsin activation, the activated collagenases appear to have the same size and activity in all of the clones. PMID- 2561463 TI - The correlation between vesicle loss and quantal secretion at the frog neuromuscular junction. PMID- 2561464 TI - Dual effect of potassium on transmitter exocytosis. AB - The time course of exocytosis of quanta of acetylcholine induced by 20 mM K+ was studied at the frog neuromuscular junction. Images of vesicle fusion on freeze fracture replicas were mostly localized at the active zones in resting preparations fixed in 20 mM K+. Fusions appeared also outside the active zones in preparations fixed after 1 min exposure to 20 mM K+ and were evenly distributed over the presynaptic membrane after 5 min in 20 mM K+ (even though secretion was prevented by withdrawing Ca2+ until 30 s before fixation). The mean densities of vesicle fusions were comparable in all conditions, as were the total number of quanta released during the fixation period. This indicates that fusions outside active zones represent ectopic exocytosis, slowly activated by potassium. Partial inactivation of K(+)-induced quantal release (time and concentration-dependent) was observed electrophysiologically; this may be related to the observed decrease in density of vesicle fusions along the active zones, with time. Consistently, after 5 min in 15 mM K+ fusion density at the active zones remained high. It is concluded that active zone-associated and ectopic fusions are two exocytotic processes activated with differential time courses and concentration-dependence by K+. PMID- 2561465 TI - Spontaneous and nerve-evoked quantal transmission in regenerated motor terminals. AB - Quantal acetylcholine release was studied in sciatic nerve-extensor digitorum longus muscle preparations from normal rats and in reinnervated muscles at different times following peroneus nerve crush. The efficiency of nerve-evoked transmission was regained almost completely at a time (20 days after crush) when the secretory response to increased extracellular K+ was very weak. This suggests that K+-induced secretion might be activated through mechanisms at least partially different from those involved in the response to nerve stimulation. PMID- 2561466 TI - Action of Ca2+ agonists/antagonists in mammalian peripheral neurons. AB - The action of several ligands on the low- (LVA,T) and high-threshold (HVA,L and N) Ca channels of adult rat sensory neurons and human neuroblastoma IMR32 cells has been investigated. In both cell types, 40 microM Cd2+ and 6.4 microM /omega Conotoxin (omega-CgTx) selectively blocked the HVA channels, sparing the majority of LVA channels that were antagonized by amiloride and Ni2+. In 50% of the cells, however, /omega-CgTx spared also a 15% of HVA channels that proved to be sensitive to BAY K 8644. The agonistic action of BAY K 8644 on [omega-CgTx resistant HVA channels caused a large Ba current increase, prolonged current deactivation and acceleration of HVA channels inactivation that was particularly evident in adult rat DRG. PMID- 2561467 TI - Cross-talk between receptors coupled to calcium currents in adult but not neonatal rat sensory neurons. AB - In adult rat sensory neurons Ca2+ currents were studied with the whole-cell patch clamp technique. Two categories of neuromodulators, known to activate different 2nd messenger systems: 1) angiotensin II (AII), bovine serum albumin (BSA), Acetylcholine (ACh) and 2) GABA, stimulated the low-voltage activated (LVA) and inhibited the high voltage activated (HVA) currents, respectively. The simultaneous application of the two types of drugs failed to inhibit the HVA current via a putative cross-talk between the two 2nd messengers. PMID- 2561468 TI - Cross-talk between different intracellular signalling pathways in the rat hippocampus. AB - The activation of alpha 1-adrenergic receptors in rat hippocampal slices enhances polyphosphoinositide (PPI) breakdown and cyclicAMP (cAMP) accumulation. The latter effect is antagonized by different protein kinase C (PKC) inhibitors and mimicked by a diacylglycerol (DAG) analogue, 1,2-diolein, which activates PKC, suggesting that cAMP synthesis is indirectly affected by alpha 1-adrenoceptors through the stimulated generation of DAG upon PPI hydrolysis. Furthermore the elevation of hippocampal cAMP decreases the ability of alpha 1-receptor agonists to enhance PPI breakdown. It is proposed that the observed effects are part of a complex cross-talk between PPI and AC signalling pathways operating in hippocampal neurons. PMID- 2561469 TI - Arachidonic acid metabolites as mediators of synaptic modulation. AB - The neurotransmitters histamine, dopamine and the peptide Phe-Met-Arg-Phe-NH2 (FMRFa) cause presynaptic inhibition in the nervous system of the marine mollusk Aplysia Californica by combined down-modulation of a Ca++ conductance and up modulation of a K+ conductance. The action of FMRFa on the S-type K+ channels of Aplysia sensory neurons is mediated by a metabolite of the 12-lipoxygenase pathway of arachidonic acid, possibly 12-HPETE. A Pertussis toxin-sensitive GTP binding protein couples FMRFa receptor to the activation of the arachidonic cascade. Once produced, 12-HPETE does not require ATP- or GTP-dependent processes to act on the K+ channels, but it may directly modulate the channel via an external membrane receptor. Based on this observation, a role for eicosanoids as possible intercellular messengers in the C.N.S. is discussed. PMID- 2561470 TI - [Evaluation of transbronchial needle aspiration in the diagnosis of bronchogenic carcinoma]. AB - Transbronchial Needle Aspiration (TBNA), utilizing the flexible fibrobronchoscope, was one of the diagnostic procedures administered to the 148 cases examined. By means of the combination of transbronchial biopsy with brushing, 110 of those 148 patients were determined to have lung cancer. Thus, the diagnostic yields of TBNA, biopsy and brushing were 70.9% (78/110) 69.09% (76/110) and 65.45% (72/110) respectively. TBNA combined with the forceps biopsy increased the diagnostic yields to 91.81% (101/110), and combined with forceps and brushing increased the diagnostic yields to 93.63% (103/110). Additionally, 38 among 148 cases with negative results in TBNA were finally proven not to have bronchogenic carcinoma. No false positive was detected. There were no serious complications in this series. Our study demonstrates that TBNA is a relatively simple, safe and effective technique in diagnosing patients suspected with lung cancer; it can be used in combination with biopsy of forceps and brushing. PMID- 2561471 TI - [Electron microscopic study of mucus secretion in gastric carcinoma and its pathologic significance]. AB - The genesis, ultrastructure and fate of mucin granule in cancer cells are described, and lysosomal degradation (crinophagy) was confirmed by demonstration of cytidine monophosphatase activity. Mucin granules usually appeared as those in the goblet cell, but were heterogenous. The chemical nature of mucin or the histogenesis of tumor should not be determined simply by depending on certain features of mucin granule, because these mucin granules might be influenced by the direction of cell differentiation, the maturity of mucin granule, and the abnormalities in its development. In this paper, different patterns of mucus secretion in various histologic types are described, and their pathologic significance is discussed. PMID- 2561472 TI - [Detection of HBV DNA in hepatocellular carcinoma by in situ hybridization using a biotin-labelled probe]. AB - Eight cases of hepatocellular carcinoma were hybridized in situ with a biotin labelled HBV DNA probe on formalin fixed paraffin embedded sections. HBV DNA was detected in 218 cases both in cancer and pericancerous tissue of the liver, and both carcinomas were well differentiated. In three cases, HBV DNA was only present in pericancerous tissue and no HBV DNA could be identified in the remaining three cases. The positive rate of HBV DNA was 25% in tumor and 62.5% in the pericancerous area of liver. HBcAg was negative in all the eight tumors, nevertheless, HBsAg was present in one case. Both HBsAg and HBcAg were positive in the peri-cancerous tissue of liver in 6 out of 8 cases. In the remaining two, one was only HBsAg positive while the other was HBcAg positive. HBV DNA was identified mainly in the cytoplasm of tumor cells. In certain cells it was seen in the perinuclear cytoplasm or beneath the nuclear envelope. Only in a few cells, HBV DNA was distributed in the nuclei diffusely. Since HBV DNA was present both in the cytoplasm and nuclei, suggesting that HBV DNA was present in either integrated or free forms. PMID- 2561473 TI - [Establishment and studies of two human pancreatic carcinoma in vivo lines by transplanting the tumor into nude mice]. AB - Two human pancreatic adenocarcinoma in vivo lines (PCT-1, PCT-2) were established in athymic nude mice and maintained for 27 passages (PCT-1) and 7 passages (PCT 2) respectively in 29 (PCT-1) and 10.5 months (PCT-2). Tumours obtained from metastatic lymph node (PCT-1) and metastatic nodule in the omentum (PCT-2) were the initial sources for implantation. All xenografts in the two tumour lines were studied by using histological, ultrastructural, immunohistochemical and cytogenetical methods. PCT-1 maintained the poorly differentiated adenocarcinoma features with little mucin production, and CEA was weakly positive with metastasis to lymph nodes in this tumour line. PCT-2 was a moderately differentiated adenocarcinoma line with abundant mucin secretion, CEA was strongly positive. The poorly differentiated tumours have a faster growth rate in comparison with those of the moderately and well differentiated ones. PMID- 2561474 TI - [Flow cytometric analysis of the effect of activated macrophages on alveolar cell carcinoma of lung in vitro]. AB - Evidences have shown that activated macrophages were cytotoxic to tumor cells but the mechanism of this killing effect is not well understood. Flow cytometry was used to study the effect of activated macrophages on cell proliferation cycle of alveolar cell carcinoma of lung (A 549). Among 74 C57BL/6J mice, 30 as experimental group were given corynebacterium parvum 0.5 ml (1 mg)/mouse intraperitoneally, and 44 as control group were given normal saline 0.5 ml/mouse instead. Peritoneal macrophages of both groups were co-cultured with A 549 in the ratio of 10:1 and 20:1 for 24 and 48 hrs respectively. Specimens were assayed with LXJ-Laser flow cytometer and relative % of tumor cells in G1/O, S and G2 + M phased were calculated by morphometry using Baisech's mathematical model and Zeiss OPTON Video plan. Results indicated that in the experimental group with E/T 20:1, co-cultured for 24 hrs., the % of tumor cells in S phase was markedly decreased (28.44%) and those in G1/0 phase was increased (60.18%) (P less than 0.01); while those of the control group were 40.41% and 49.99% respectively. Changes present in S and G1/0 phase tumor cells of the experimental group indicate that activated macrophages may block tumor cells from the G1/0 to S phase and thus affect their proliferation. PMID- 2561475 TI - [Abnormal inositol phospholipid metabolism as a main factor causing pericyte drop out in diabetic retinopathy]. AB - The biochemical basis of pericyte loss in early diabetic retinopathy is still an open question. In studies on the mechanism by which retinal pericytes degenerate, we first established a bovine retinal capillary pericyte (BRCP) cell line. Subcultured BRCP grown in high (10-40 mmol/L) glucose media were used as an experimental model. We found that high concentrations of glucose suppress the mitotic rate and cell birth rate of cultured BRCP. High concentrations of glucose inhibit myo-inositol transport and result in decreased intracellular myo-inositol content. This inhibition can be partially reversed by sorbinil, an aldose reductase inhibitor (ARI). Myo-inositol is a precursor of inositol phospholipids (IPLs), whose metabolism is responsible for a number of signal transduction processes. Phosphoinositidase (PIase) cleaves the phosphodiester bond of phosphotidylinositol 4,5 diphosphate (PIP2) to produce two second messengers, inositol trisphosphate (IP3) and diacylglycerol (DG). Further experiments showed that IP3 and DG synergistically activate BRCP proliferation in vitro. High concentrations of glucose altered the formation of both IPLs and inositol phosphate esters (IPEs) in an organ culture of retinal microvessels. This alteration can be reversed by adding either high concentrations of myo-inositol or ARI to the medium. PIase activity was attenuated to 82% or 55% when glucose in the growth medium was increased from 5 to 15 or 30 mmol/L, respectively. When IPLs from BRCP were analyzed by HPLC and TLC, we observed the reduction of three IPLs, including the substrate of PIase, PIP2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561476 TI - [Morphological study of Omsk hemorrhagic fever (OHF) virus with the electron microscope]. AB - Morphological characterization of OHF virus was studied by means of thin section electron microscopy in the brains of mice infected with Omsk hemorrhagic fever virus. The crystals, microvesicles, vacuoles and as beehive structures were observed in the virus-infected cells. The possible relationship between the characteristic structures and virus morphogenesis is discussed. It is indicated that these characteristic structures may be important to morphological characterization of OHF virus. PMID- 2561477 TI - [Investigation of the effect of a diet with wheat bran on the metabolic balances of Zn, Cu, Ca and Mg in diabetics]. AB - We studied the effect of diet with wheat bran for 4 weeks on the metabolic balances of Zn, Cu, Ca and Mg in 10 diabetics who had been well-controlled, and compared with 10 healthy persons and 14 uncontrolled diabetics. The amount of wheat bran used was 0.4 g/kg/day. Our results showed that wheat bran could lower blood glucose, increase the intestinal absorption and positive balance of Zn, whereas no effect on the metabolic balances of Ca, Cu and Mg. It was concluded that diabetic diet adding appropriate wheat bran was helpful for diabetic control and the correction of Zn deficiency. PMID- 2561478 TI - [Endothelin--a newly discovered vasoactive peptide]. PMID- 2561479 TI - The role of retinoid-binding proteins in the generation of pattern in the developing limb, the regenerating limb and the nervous system. AB - We summarize existing data and describe new information on the levels and distribution of cellular retinoic acid-binding protein (CRABP) and cellular retinol-binding protein (CRBP) in the regenerating axolotl limb, the developing chick limb bud and the nervous system of the chick embryo in the light of the known morphogenetic effects of retinoids on these systems. In the regenerating limb, levels of CRABP rise 3- to 4-fold during regeneration, peaking at the time when retinoic acid (RA) is most effective at causing pattern duplications. The levels of CRBP are low. The potency of various retinoids in causing pattern respecification correlates well with the ability of these compounds to bind to CRABP. In the chick limb bud, the levels of CRABP are high and the levels of CRBP are low. Again the binding of various retinoids to CRABP correlates well with their ability to cause pattern duplications. By immunocytochemistry, we show that CRABP is present at high levels in the progress zone of the limb bud and is distributed across the anteroposterior axis in a gradient with the high point at the anterior margin. In the chick embryo, CRABP levels are high and CRBP levels are low. By immunocytochemistry, CRABP is localised primarily to the developing nervous system, labelling cells and axons in the mantle layer of the neural tube. These become the neurons of the commissural system. Also sensory axons label intensely with CRABP whereas motor axons do not and in the mixed nerves at the brachial plexus sensory and motor components can be distinguished on this basis. In the neural tube, CRBP only stains the ventral floor plate. Since the ventral floor plate may be a source of chemoattractant for commissural axons, we suggest on the basis of these staining patterns that RA may fulfill this role and thus be involved morphogenetically in the developing nervous system. PMID- 2561480 TI - Custom made fabricated porous hydroxyapatite block implants for augmentation of partially edentulous atrophic alveolar ridge. PMID- 2561481 TI - [Papillomas of the oral cavity. Role of the papillomavirus]. AB - Latest results about etipatogenesis of oral papillomas make us to modify the old classification according which papillomatosis were traumatic or idiopathic, giving to the latest a tested viral origin. Studies at the electron microscopy, immunoperoxidase technique and DNA-hybridization showed that human papillomavirus (HPV) is the etiologic agent of these oral lesions. Many works seem to support the opinion that HPV could be implicated even in the development of some oral carcionomas; anyway, further confirmatory evidence is still needed. PMID- 2561482 TI - [Human papilloma virus and pathology of the oral cavity]. AB - The human papillomavirus (HPV) causes a variety of cutaneous warts and proliferative squamous lesions of oral mucosa. This article reviews, on the basis of the literature and personal observations, the biology of HPV and its etiopathogenetic role in some uncommon oral diseases. PMID- 2561483 TI - Medullomyoblastoma. A case report. AB - A case of medullomyoblastoma was studied by light and electron microscopy and by immunohistochemistry. It showed glial and neuronal differentiation in the medulloblastoma areas and rhabdomyoblastic differentiation in the intervening areas. PMID- 2561484 TI - Formation and effects of leukotriene B4 in human lymphocytes. AB - Incubation of human tonsillar B lymphocytes or peripheral blood T lymphocytes with leukotriene (LT) A4 led to the formation of LTB4. Stimulation of these cells with ionophore A23187 did not lead to the synthesis of detectable amounts of leukotrienes. Formation of LTB4 was observed in several monoclonal B- and T-cell lines after incubation with LTA4, but not after stimulation with ionophore A23187. The Burkitt lymphoma cell line Raji was found to possess higher LTA4 hydrolase activity than normal lymphocytes. The expression of the LTA4-hydrolase gene but not the 5-lipoxygenase gene was demonstrated on the transcriptional level in Northern blots and on the translational level by Western blots. Stimulation of human monocytes with ionophore A23187 resulted in the release of LTA4. Coincubations of transformed lymphocytes and monocytes stimulated with ionophore A23187 produced increased amounts of LTB4 as compared with monocytes alone. LTB4 influence on lymphocyte activation was studied and CD23 expression was used as a marker. The expression of this antigen was enhanced on resting B lymphocytes in synergy with B-cell growth-promoting factors. LTB4 also augmented DNA synthesis, cell replication and IgG secretion. These results indicate that extracellular LTA4, released from activated monocytes, is converted by lymphocytes into LTB4 which might cause activation and differentiation of B lymphocytes. PMID- 2561485 TI - Anterior pituitary functions in patients with uremia tested by stimulation with four combined hypothalamic releasing hormones. AB - Seven cases with uremia (6 men, 1 woman, mean age = 55.6 +/- 2.2 years) were studied with four combined hypothalamic releasing hormones (corticotropin releasing hormone, CRH; luteinizing hormone-releasing hormone, LHRH; thyrotropin releasing hormone, TRH; and growth hormone-releasing hormone, GHRH) for assessment of anterior pituitary functions. The mean basal levels of corticotropin (ACTH, 22.4 +/- 5.2 pg/ml), thyrotropin (TSH, 2.4 +/- 0.6 microU/ml), and follicle stimulating hormone (FSH, 26.0 +/- 3.4 mIU/ml) in uremic patients were not significantly different from those (34.0 +/- 3.5 pg/ml, 2.0 +/- 0.4 microU/ml, and 23.2 +/- 6.4 mIU/ml) of controls (5 men, 1 woman, mean age = 54 +/- 2.5 years), but the ACTH and TSH responses to the releasing hormones were significantly lower than those of the controls. The mean basal levels of luteinizing hormone (LH, 70.7 +/- 16.3 mIU/ml), cortisol (9.8 +/- 1.2 micrograms/dl) and prolactin (109.3 +/- 23.2 ng/ml) in uremic patients were significantly higher than those of normals (27.3 +/- 6.6 mIU/ml, 6.5 +/- 0.7 micrograms/dl and 15.7 +/- 3.4 ng/ml), while suppressed LH, cortisol and prolactin responses to the releasing hormones were observed in the uremic group. The mean basal growth hormone (GH) level in uremic patients (3.1 +/- 0.4 ng/ml) was not significantly different from that (2.8 +/- 0.7 ng/ml) of normals, but the GH response to the releasing hormones was significantly higher than that of controls. These results show pituitary dysfunction, such as blunted ACTH, TSH, LH and prolactin response, exists in uremic patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561486 TI - Cytomegalovirus retinitis treated with ganciclovir in a renal transplant recipient. AB - Cytomegalovirus retinitis is an opportunistic and vision-threatening disease in immunosuppressed patients. Currently available antiviral agents have been unsuccessful in halting its progression. We report a case of cytomegalovirus retinitis which occurred 6 months after renal transplantation and was successfully treated with an investigational drug, ganciclovir (DHPG [Dihydroxy propoxymethyl guanine], Cytovene, BW-B759U), via intravenous administration, with resolution of active retinal lesions and the cessation of cytomegalovirus shedding in the urine. There was neither a recurrence of cytomegalovirus retinitis nor an occurrence of retinal detachment during the 9-month follow-up after intravenous ganciclovir therapy. PMID- 2561487 TI - Streptomyces griseus streptomycin phosphotransferase: expression of its gene in Escherichia coli and sequence homology with other antibiotic phosphotransferases and with eukaryotic protein kinases. AB - The aphD gene of Streptomyces griseus, encoding a streptomycin 6 phosphotransferase (SPH), was sub-cloned in the pBR322-based expression vector pRK9 (which contains the Serratia marcescens trp promoter) with selection for expression of streptomycin resistance in Escherichia coli. Two hybrid plasmids, pCKL631 and pCKL711, were isolated which conferred resistance. Both contained a approximately 2 kbp fragment already suspected to include aphD. The properties of in vitro deletion derivatives of these plasmids were consistent with the presumed location of aphD. In vitro deletion of a sequence including most of the trp promoter largely, but not quite completely, abolished the ability of the plasmid to confer streptomycin resistance, confirming that expression was indeed principally from the trp promoter. A polypeptide of approximately 34.5 kDa was present in minicells containing plasmids that conferred streptomycin resistance, but was absent when the plasmids contained in vitro deletions removing streptomycin resistance. Part of the fragment was sequenced and an open reading frame corresponding to aphD identified. A computer-assisted comparison of the deduced SPH sequence with those of other antibiotic phosphotransferases suggested a common structure A-B-C-D-E, where B and D were conserved between all sequences compared while A, C and E divided between the streptomycin and hygromycin B phosphotransferases on one hand and kanamycin/neomycin ones on the other. A composite sequence data base was searched for homologues to consensus matrices constructed from five approximately 12-residue subsequences within blocks B and D. For one subsequence, corresponding to the N-terminal portion of block D, those sequences from the database that yielded the highest homology scores comprised almost entirely either antibiotic phosphotransferases or eukaryotic protein kinases. Possible evolutionary implications of this homology, previously described by other groups, are discussed. PMID- 2561488 TI - Purification and properties of NADP-dependent glutamate dehydrogenase from Streptomyces fradiae. AB - Streptomyces fradiae has two chromatographically distinct forms of glutamate dehydrogenase (GDH): one GDH utilizes NAD as coenzyme, the other uses NADP. The intracellular level of both GDHs is strongly regulated by the nitrogen source in the growth medium. NADP-dependent GDH was purified to homogeneity from crude extracts of S. fradiae. The Mr of the native enzyme was determined to be 200,000 by size-exclusion high-performance liquid chromatography whereas after sodium dodecyl sulphate-polyacrylamide gel electrophoresis one major band of Mr 49,000 was found, suggesting that the enzyme is a tetramer. The enzyme was highly specific for the substrates 2-oxoglutarate and L-glutamate, and required NADP, which could not be replaced by NAD, as a cofactor. The pH optimum was 9.2 for oxidative deamination of glutamate and 8.4 for reductive amination of 2 oxoglutarate. The Michaelis constants (Km) were 28.6 mM for L-glutamate and 0.12 mM for NADP. Km values for reductive amination were 1.54 mM for 2-oxoglutarate, 0.07 mM for NADPH and 30.8 mM for NH+4. The enzyme activity was significantly reduced by adenine nucleotides, particularly ATP. PMID- 2561489 TI - Transfer and properties of some natural and suicide replicons in Pasteurella multocida. AB - We tested the transfer of several plasmids and transposons from Escherichia coli to Pasteurella multocida by filter mating. Two plasmids, pRKTV5 (pRK2013::Tn7) and pUW964 (pRKTV5::Tn5), were derived from pRK2013--a narrow-host-range plasmid with the broad-host-range IncP conjugation genes. Most P. multocida transconjugants obtained with pRKTV5 had Tn7 insertions in the chromosome but some had insertions of the whole plasmid. By contrast, all the transconjugants obtained with pUW964 had insertions of this plasmid or a deleted variant. pUW964 mediated low-frequency transfer of Tn7 or chromosomal markers between P. multocida strains. Broad-host-range IncP plasmid RP4 (RK2) did not yield selectable transconjugants in P. multocida but two plasmids derived by Tn5 insertion into a kanamycin-sensitive derivative of RP4 did yield transconjugants. pSUP1011, a narrow-host-range p15A replicon with the RP4 mob region allowing mobilization by the IncP conjugation genes also yielded transconjugants while several other plasmids tested did not transfer markers to P. multocida. PMID- 2561490 TI - [Polyagglutinability in erythrocytes]. AB - A case of erythrocyte polyagglutinability is presented which developed in a patient who underwent surgery in the course of infection with Clostridium perfringens. When blood transfusion became necessary it was decided that the patient be transfused with concentrated and washed erythrocytes. The transfusion was well tolerated. With the curing of the infection the polyagglutinability also disappeared. PMID- 2561491 TI - [Distal tubular acidosis. Description of a classic case and of a type 4 case in neonatal single kidney]. AB - During the last two years two babies with failure to thrive developed distal renal tubular acidosis (RTA). We report a female patient with classic RTA where the failure to thrive occurred from the 8th month of age associated with polyuria and polydipsia. A 45 day old infant had RTA type 4 associated with unilateral renal agenesis; type 4-RTA is characterized by inadequate urinary potassium excretion in the face of hyperkalemia, in distinct contrast to the other type of RTA in which enhanced kaliuria is observed. Treatment with oral bicarbonate resulted in sustained normalization of blood acid-base status and accelerated linear growth in both babies. PMID- 2561492 TI - Cationic anticancer drugs and their modes of action. AB - This report focuses on two groups of cationic cancerostatics, anthracycline antibiotics and 1,4-benzoquinone-guanylhydrazone-thiosemicarbazone (ambazone), lining up biophysical and biochemical effects on the level of membranes and membrane constituents. The interaction of both drugs with multilamellar liposomes consisting of phosphatidylcholine used as a simple model membrane system could be ensured by means of steady state and nanosecond time-resolved fluorometric investigations. The biochemical effect on membranes is underlined by the inhibition of the neuraminidase activity of the Sendai virus, modification of the CAMP phosphodiesterase activity of leukemia L 1210 cells of mice and reduction of the lymphocyte blast transformation. PMID- 2561493 TI - [Primary pulmonary cancer in women the the Doboj Region]. AB - The aim of this work is primary lung cancer in female population of the Doboj district in the period 1975-88. incidence research as well as age structure, localisation, histological types, stage and diseases associated with lung cancer. A group of 114 female patients was investigated and as comparative group male patients with lung cancer in the same period was used. The results show that incidence rate increase is significant and in the future that trend will be continued. Especially serious is often microcellular and adenocarcinoma appearance as well as female non-smoking patients (42%) and patients with diabetes mellitus (18%) in the investigated group high percentage. These phenomena lead to conclusion that risk factors are partly different and they should be better investigated. Females with diabetes mellitus require special treatment and further research should be focused on diabetes--lung cancer relationship. PMID- 2561494 TI - Changes in milk carbohydrates during lactation in the common brushtail possum, Trichosurus vulpecula (Marsupialia:Phalangeridae). AB - Milk samples (186) were obtained at various stages of lactation from 27 common brushtail possums (Trichosurus vulpecula). Qualitative and quantitative changes in the milk carbohydrates during early and mid-lactation were similar to those previously seen in other marsupials; the principal carbohydrate was lactose early in lactation and higher oligosaccharides in mid-lactation, and the hexose concentration reached a peak during mid-lactation. However, the late-lactation milk was unusual in that the carbohydrate was mainly lactose and its concentration remained relatively high (3.5 to 5.5%). In contrast to earlier findings on the milk of the tammar wallaby (Macropus eugenii), little or no nucleotide pyrophosphatase, beta-galactosidase and alkaline phosphatase activities were detected late in lactation. PMID- 2561495 TI - Cortisol feedback on ovine fetal ACTH secretion during the last fifth of gestation. AB - This study investigates the development of the feedback relationship between cortisol and ACTH in ovine fetuses at 117-119 (n = 5), 130-134 (n = 4) and 140 143 (n = 6) days of gestation. During the last 2 h of a 24 h cortisol (100 micrograms h-1) or saline (0.19 mL h-1) infusion, the fetal ACTH response to bolus oCRH injection (10 micrograms) was tested after the collection of basal ACTH and cortisol samples. Basal ACTH concentrations in un-infused or saline infused fetuses were observed to progressively increase after 117-119 days of gestation. In contrast, a prepartum increase in cortisol concentrations could not be detected until after 130-133 days. Cortisol infusion significantly inhibited basal ACTH values at 130-133 and 140-143 but not at 117-119 days. In the two youngest age groups, cortisol infusion significantly inhibited the fetal ACTH response to oCRH. At 140-143 days, ACTH values after oCRH injection, were elevated to a similar extent during the saline or cortisol infusion. However, at 140-143 days there was evidence to suggest that the ACTH response to oCRH administered during the saline infusion was blunted when compared with earlier stages of gestation. This study suggests that the circulating, basal concentrations of both ACTH and cortisol increase during the last fifth of gestation in the ovine fetus. The exact nature of the prepartum feedback relationship that develops between these two hormones remains to be fully elucidated. PMID- 2561496 TI - Proteolytic catabolite inactivation in Saccharomyces cerevisiae. AB - Fermentable sugars, when added to cells of Saccharomyces cerevisiae growing on a non-fermentable carbon source, cause repression of the synthesis of certain enzymes ("catabolite repression") and in addition inactivation of a smaller group of enzymes ("catabolite inactivation"). Enzymes for which "catabolite inactivation" has been observed are listed herein. In five cases, it has been shown that the mechanism of catabolite inactivation is proteolytic in nature. Our present knowledge on the conditions and the mechanisms of initiation of inactivation and the biological significance of the proteolytic inactivation is summarized for these five enzymes: cytoplasmic malate dehydrogenase, aminopeptidase I, fructose-1,6-bisphosphatase (FBPase), phosphoenolpyruvate carboxykinase and isocitrate lyase. With the exception of aminopeptidase I, these enzymes are key enzymes of gluconeogenesis in S. cerevisiae. It is obvious that gluconeogenesis is no longer necessary, if a fermentable carbon source is available. PMID- 2561497 TI - Intracellular degradation of newly synthesized collagen. AB - This chapter reviews recent work on intracellular degradation of newly synthesized collagen, the most abundant protein in the mammalian body. Approximately 10-20% of collagen synthesized by human fibroblasts under normal culture conditions is broken down rather than secreted; this is referred to as the basal level of degradation. It is not known where basal degradation occurs, but indirect evidence suggests a compartment of the secretory pathway, perhaps the Golgi complex. Intracellular degradation is increased when cells are induced to synthesize structurally abnormal collagen, either by incubation in the presence of amino acid analogs or because of mutations in collagen genes. There is evidence that lysosomal proteases mediate degradation of structurally abnormal collagen, however, recent work indicates that I-cells, which are genetically deficient in several lysosomal enzymes, can also degrade abnormal collagen. Modulation of the level of intracellular degradation can effect a change in net collagen production independently of changes in the rate of synthesis. Intracellular degradation of newly synthesized collagen has been observed in vivo, but it is not clear whether decreasing degradation contributes to increasing collagen production in fibrosis. This chapter concludes by relating collagen degradation to the more general phenomenon of intracellular degradation of newly synthesized secretory proteins, and by indicating directions for future work. PMID- 2561498 TI - Selective aspects of mitochondrial protein turnover. AB - To establish the interrelationship between protein synthesis and degradation, we have developed a simple procedure. We have chosen the mitochondrial proteins, because most of them are synthesized outside mitochondria and are then imported into the organelle. This fact allows to separate easily the synthesis steps from the general turnover. By using this system we have some evidences suggesting that the in vitro protein concentration in mitochondria may be regulated by the entry of mitochondrial protein precursors. Also, we are studying the factors that regulate the import of mitochondrial protein precursors into the organelle, because this step may be essential in the regulation of the turnover of mitochondrial proteins. PMID- 2561499 TI - ATP-dependent proteolysis of mitochondria of reticulocytes. AB - Since its discovery in 1956 the physiological function of the ATP-dependent proteolysis in reticulocytes has become an area of intensive interest. The evidence that mitochondria are the main substrate of the process during maturation of reticulocytes is summarized. The large consumption of ATP related to the degradation of mitochondria is discussed in context with possible mechanisms for ATP requirement. A novel ATP-dependent peptide release from reticulocyte mitochondria, which may be the first step in the degradation of the organelles, is demonstrated. Ubiquitin and tRNA are involved in the degradation process. Their roles, however, are not yet clear. The whole degradation cascade consisting of lipoxygenase, mitochondria susceptibility factor and ATP-dependent proteolysis, may function as a defense system against uncontrolled degradation of mitochondria in reticulocytes. PMID- 2561500 TI - Suppression of testicular activity by a GnRH agonist in hypophysectomized, gonadotropin-treated mice. AB - Although it is believed that the suppressive effect of GnRH agonist on the testes in mediated, at least in part, through its direct action on the testes in rats, such direct action was considered to appear in rats alone and in no other species, including mice, primates and humans. To reexamine such species' specific direct action of GnRH agonists, the effect of a potent GnRH agonist, D-Leu6-GnRH (GnRH-A), on testicular activity was investigated in hypophysectomized, gonadotropin-treated mice. Thirteen days after hypophysectomy the testicular weight had decreased to 20% of that of intact mice. Daily injection of 1 IU of HCG and 0.1 microgram of HMG for 10 days restored the testicular weight to 50% of that of the control animals. Daily sc injections of 1, 0.1 or 0.01 micrograms of GnRH-A (in addition to HCG and HMG) slightly but significantly prevented restoration of testicular weight by exogenous gonadotropins in hypophysectomized mice. A more dramatic effect of GnRH-A was observed in serum testosterone levels. Hypophysectomy resulted in a nearly 96% reduction in serum testosterone level. Treatments with HCG and HMG raised the levels to those found in intact mice. Daily injections of GnRH-A resulted in a marked reduction of testosterone levels at all doses of GnRH-A tested to those corresponding to hypophysectomized, vehicle-treated animals. Daily injection of 1 microgram of GnRH-A also reduced both testicular weight and testosterone levels, but the magnitudes of these reductions were smaller than those observed in hypophysectomized, gonadotropin treated mice. The results clearly indicated that chronic treatment with GnRH-A reduces testicular function through direct action in mice under the experimental conditions employed. PMID- 2561501 TI - Suppression of deoxyguanosine cytotoxicity and deoxyguanosine kinase activity in mouse FM3A mammary carcinoma cell: mutants defective in hypoxanthine phosphoribosyl-transferase. AB - The cytotoxic effect of deoxyguanosine was examined in various cell clones isolated from cultured mouse FM3A mammary carcinoma cells. The inhibitory effect of deoxyguanosine on the growth of the wild-type cell was suppressed by the addition of hypoxanthine to the culture medium. Cell mutants defective in hypoxanthine phosphoribosyl-transferase (Hprt- mutants) were approximately 20 times less sensitive to deoxyguanosine. But the sensitivity of the Hprt+ revertants was restored and has become close to that of the wild-type cell. Both uptake and incorporation of [3H]-hypoxanthine and [3H]-deoxyguanosine did not occur in the cellular acid-soluble and insoluble fractions in Hprt- mutants but they were restored in all Hprt+ revertants. When the activities of deoxyguanosine kinase were measured immediately after preparation of the cell-free extracts from the Hprt- mutants, it was negligible but activity comparable to that of the wild type appeared after dialysis of the extract. These results are explained by assuming accumulation of purine metabolite in Hprt- mutants which inhibited the deoxyguanosine salvage pathway. PMID- 2561502 TI - [The results of research conducted at the Stefan S. Nicolau Institute of Virology on the herpes simplex virus]. AB - The report is a review of the research and results of the "Stefan S. Nicolau" Institute of Virology scientists in the field of herpes viruses, as they were materialized in the great number of published or communicated scientific papers these last ten years. PMID- 2561503 TI - Viral infection correlated with superoxide anion radicals production and natural and synthetic copper complexes. AB - Studies conducted on asymmetric triazine derivatives synthetized at the Chemical and Pharmaceutical Research Institute showed that products S1, S16, S17, S19, S20 and S22 have a remarkable O2- radical scavenger activity. Among these derivatives, the product S1 is the most efficient as an antiviral agent. PMID- 2561504 TI - The thyroid as a model endocrine system. PMID- 2561505 TI - Characterization of receptors for insulin and insulin-like growth factor-1 on FRTL-5 thyroid cells. AB - FRTL-5 rat thyroid cells have receptors for both insulin and IGF-I which can be distinguished in binding studies. The ability of TSH to regulate each in an antiparallel manner is atypical. If these receptors are shown to have independent as well as coordinate activities, studies of the mechanisms of their receptor cross-talk in these cells will be relevant to understanding IGF-I and insulin receptors in other tissues. PMID- 2561506 TI - Receptors of the thyroid: the thyrotropin receptor is only the first violinist of a symphony orchestra. AB - A basic reason for undertaking these studies was to further our knowledge of the structure and function of the TSH receptor as well as its interaction with other receptors on thyroid cells. The multiplicity of observations suggests the approach is bearing fruit, does not provide a simple answer, and can have pitfalls. We hope they may also contribute to understanding the structure and function of autoantigens in Graves' disease and glycoprotein hormone receptors in general. The authors are grateful to their collaborators in the National Dental Institute, particularly Drs. Bellur Prabhakar, Edward Oates, and Abner Notkins, in the National Cancer Institute, Drs. W. O. McBride and M. Lerman for their contributions to the cloning studies. PMID- 2561507 TI - G protein-linked receptors in the thyroid. AB - The FRTL5 cell line has the advantage that its hormonal activation leads to important and measurable thyroid function such as the transport of iodide and the iodination of thyroglobulin. Secondly, the coexistence in the plasma membrane of these cells of several physiologically relevant receptors (TSH, alpha 1 adrenergic, M1 and M2 muscarinic, insulin, IGF1) coupled to at least three transducing enzymes (adenylyl cyclase, PLC, PLA2) gives the possibility to analyze the interaction among second messengers in the cell activation process. This has allowed us and others to show that in the case of the iodide efflux regulation at least two second messengers (Ca++ and arachidonic acid) mediate the adrenergic stimulation, whereas the TSH activation of the same phenomenon probably uses other signals in addition to Ca++ and arachidonic acid. Growth is mostly regulated by TSH, that activates the adenylyl cyclase by a mechanism that may involve the modulation of the availability of Gi. TSH is also able to regulate an endogenous ADP ribosyl transferase in FRTL5. This could be a novel mechanism of cell regulation by this hormone, but the role of this phenomenon in the physiological action of TSH is still unclear. PMID- 2561508 TI - ADP ribosylation and G protein regulation in the thyroid. PMID- 2561509 TI - The inositide and arachidonic acid signal system. PMID- 2561510 TI - [Effect of cytoxan on immune response of Japanese encephalitis virus killed and live vaccines]. AB - Mice vaccinated by Japanese encephalitis (JE) virus killed and live vaccines respectively were injected subcutaneously with cytoxan (1.5 mg/10 g weight) once every two days for 3 times, and then challenged by JE virus virulent A2 strain. The results showed that the protective rate of killed vaccine (inoculated 3 times) was decreased from 100% to 20% after cytoxan immunosuppression, while that of live vaccines 2-8 strain and 5-3 strain (inoculated 2 times) still remained 100% as in the control group without cytoxan. The results indicate that the immune response induced by the killed vaccine can be easily damaged by cytoxan, but that induced by the live vaccines not, suggesting of important advantage of JE virus live vaccine. Our results implies that the JE incidence being higher in old subjects probably is related to the declining of immune function. PMID- 2561511 TI - [Molecular basis of pharmacological action of irreversible opioid receptor agonist alpha-CAO]. AB - Molecular mechanism of opioid effect was studied in mouse brain using alpha-CAO (7 alpha-N, N-Bis (beta-chloroethyl)amino-6, 14-endo-ethenotetrahydrooripavine), an irreversible opioid receptor agonist. There was a biphasic response in cerebral cAMP content, including an initial sharp decline and a subsequent increase 3 days later, the response being in line with analgesic effect initiated by administration of alpha-CAO and development of habituation due to sustained drug administration. Further laboratory study indicated that in the acute stage, adenylate cyclase activity in the mouse brain was inhibited by 48% and the inhibitory effect was reversible by naloxone. The delayed effect of alpha-CAO in development of habituation was accompanied by increase of AC activity and cAMP content of the brain, calmodulin content in the brain. The later being also causative factor in development of habituation. PMID- 2561512 TI - [Using BPV transforming foci as selective markers to express HBsAg]. AB - We constructed a plasmid pBMTHBR2 containing mouse MT promoter, entire HBsAg gene (preS and S gene), splicing signal from SV40 and complete BPV genome. Using calcium phosphate precipitation technique to transform C127 Cells and using BPV transforming foci as selective markers, we obtained transformed cell clones. The experiment shows that 57% of the cloned cell lines can secrete HBsAg continuously. PMID- 2561513 TI - [Expression of the Epstein-Barr virus P150 viral capsid antigen in Escherichia coli for the use as antigen in diagnostic tests]. AB - We have attempted to produce the P150 viral capsid antigen of Epstein-Barr Virus (EBV) in Escherichia coli. This protein was found, by immunoprecipitation, to be a clinically relevant antigen, especially for the determination of the IgA-titer in patients with Naso Pharyngeal Carcinoma (NPC). Since the expression of the entire P150 coding region was unsuccessful, we synthesized only the antigenic parts of this protein. After purification of P150 expressed by PUR290CXH580 with column chromatography (sepharose 2B-CL), the resulting product reacts particularly well with IgA antibodies of NPC patients indicating its diagnostic value for NPC. PMID- 2561514 TI - Effect of dietary protein intake and angiotensin converting enzyme inhibition in Heymann nephritis. AB - The effect of diets containing 8.5%, 21% or 40% protein on growth, urinary albumin excretion and serum albumin concentration was determined in rats with Heymann nephritis and in non-nephrotic control animals. Urinary albumin excretion was greater in nephrotic rats with each increment in dietary protein intake, and serum albumin concentration tended to be least in nephrotic rats fed 40% protein. Albuminuria decreased spontaneously and serum albumin concentration increased in nephrotic rats fed 8.5% protein for 25 days. Enalapril treatment caused a further reduction in urinary albumin excretion and an increase in serum albumin concentration in nephrotic rats fed 8.5% protein. Albuminuria did not decrease nor did serum albumin concentration increase in nephrotic rats fed 40% protein without enalapril treatment, but enalapril caused a significant reduction in urinary albumin excretion and an increase in serum albumin concentration in nephrotic rats fed either 8.5% or 40% protein. The rate of growth in normal rats was greatest when they were fed 21% protein, compared to either 8.5% or 40% protein. Growth rate was significantly reduced in nephrotic rats, regardless of dietary protein intake and regardless of treatment with enalapril, but the 21% protein diet still induced the most rapid rate of growth. Growth rate was not significantly different in nephrotic rats fed either 40% or 8.5% protein. The difference in weight between pair fed nephrotic and control animals fed 21% protein was due to a decrease in carcass and skin weight in nephrotic animals. Carcass protein was significantly reduced in the nephrotic animals, and carcass saponafiable fat tended to be reduced.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561515 TI - Effect of dietary protein restriction and angiotensin converting enzyme inhibition on protein metabolism in the nephrotic syndrome. PMID- 2561516 TI - High protein diets stimulate albumin synthesis at the site of albumin mRNA transcription. AB - High dietary protein intake directly stimulates albumin synthesis ald albuminuria in rats with Heymann nephritis. Increased albumin synthetic rate might be due to the presence of increased amino acids available for protein synthesis causing more efficient translation of preformed albumin mRNA, or instead might be linked to increased steady state albumin mRNA levels in the liver. Albumin synthesis, hepatic albumin mRNA content, and albumin mRNA relative to beta actin mRNA (as an internal control) (Alb/beta Act), were measured in rats with Heymann nephritis fed either 8.5% protein (LP), or after protein intake was increased to 40% for 4 days (HP). enalapril (E) was used to modulate the proteinuric effect of HP, yielding four experimental groups, LPN (8.5% protein nephrotic, no enalapril), LPE (8.5% protein, enalapril treated), HPN (40% protein nephrotic, no enalapril), and HPE (40% protein, enalapril treated). Dietary protein augmentation increased the rate of albumin synthesis, steady-state albumin mRNA levels, and Alb/beta Act in both HPN and HPE, compared to either LPH or LPE, even though serum albumin concentration was greater in HPE than in either of the groups fed LP. Both albumin mRNA and Alb/beta Act correlated with the rate of albumin synthesis (r = 0.531, P less than 0.05; and 0.553, P less than 0.01 respectively). Nuclear run on assays were performed using nuclei isolated from the livers of LPN or HPN to determine whether increased albumin mRNA resulted from an increase in the rate of albumin mRNA transcription.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561517 TI - [Modern aspects of diagnosis and therapy of Wilm's tumor]. AB - Real-time sonography of 12 cases with Wilms' tumour confirmed by the pathohistological finding was presented according to the ultrasonographic picture and the possibilities of radiotherapeutic treatment in dependence of the degree of disease. Uniform, irregularly reflective appearance of the tumourous tissue was observed in 58.3% of the patients, while the areas of necrosis and/or hemorrhage and cystic degeneration in the tumour accounted for 41.7%. Clear determination between the tumour and the normal parenchyma of the kidney was confirmed in 66.6% of the cases, while visualization of the normal tissue of the kidney was not possible in 25% of the patients. The treatment of patients with Wilms' tumour was narrowly coordinated by the program consisting of the surgical extirpation of the tumour, postoperative irradiation of the tumorous area at degrees II, III, IV and V and intensive adjuvant chemiotherapy. PMID- 2561518 TI - [Sequential infusion of GHRH 1-29NH2, LHRH and TRH for evaluating the hypophyseal reserve. Comparison with Politest]. AB - Fourteen patients with pituitary lesions and 6 normal subjects underwent the following tests: a) LHRH (100 mcg) + TRH (200 mcg) + Human Insulin (0.1 UI/kg) i.v. (Politest "A"); b) LHRH (100 mcg) + TRH (200 mcg) + GHRH 1-29NH2 (100 mcg) i.v. (Politest "B"); c) GHRH 1-29NH2 (100 mcg) i.v. (GHRH-TEST). FSH, LH and PRL, as assayed in patients and controls in response to Politest "A" and "B", revealed no significant differences between the tests. Plasma TSH values were significantly higher after Politest "B" in both patients and normals. The GH response after Politest "A" and "B" was similar in magnitude and concordant in 88.8% of cases. PMID- 2561519 TI - Impairment of olfactory memory by local infusions of non-selective excitatory amino acid receptor antagonists into the accessory olfactory bulb. AB - Female mice form a long-term olfactory memory to the pheromones of the male that mates with them. This memory is dependent on neural mechanisms within the accessory olfactory bulb. In this study we show that localized infusions of the excitatory amino acid receptor blocker, gamma-D-glutamylglycine, into the accessory olfactory bulb prevents memory formation. This is in marked contrast to the effects of infusions of the specific N-methyl-D-aspartate receptor antagonists, D-2-amino-5-phosphonovaleric acid and MK 801, which are without effect on memory formation. Excitatory amino acid receptor blockade by localized infusion of these drugs into the accessory olfactory bulb induced seizures. This paradoxical effect could only be due to disinhibition of granule cell GABAergic inhibitory feedback to the mitral cell. This was confirmed by the pregnancy blocking effect of these drugs, an event which also occurs with bicuculline infusions into the accessory olfactory bulb. These findings strongly implicate excitatory amino acid receptors in memory formation to the pheromones of the mating male and localize the mechanism to the reciprocal dendro-dendritic synapse between mitral and granule cells. PMID- 2561520 TI - Anatomically distinct output channels of the caudate nucleus and orofacial dyskinesia: critical role of the subcommissural part of the globus pallidus in oral dyskinesia. AB - The findings in this feline study indicate that the enkephalin-positive subcommissural part of the globus pallidus, which is known to contain GABA and cholinergic cells projecting to the cortex, is innervated by the anterodorsal region of the caudate nucleus, but not by the core. Like stimulation of a particular subclass of dopamine receptors in the anterodorsal region of the caudate nucleus, inhibition of the GABA receptors in the noted part of the globus pallidus resulted in orofacial dyskinesia, viz. tic-like contractions of the facial, eye and ear muscles, and tongue protrusions. This phenomenon was elicited by intrapallidal injections of the GABA antagonist picrotoxin in a dose-dependent manner and could be attenuated by the GABA agonist muscimol. Previous studies have already shown that neither stimulation of the dopamine receptors in the core of the caudate nucleus nor any manipulation with the first- and second-order output-stations of the latter brain region, viz. (a) those regions of the substantia nigra, pars reticulata which receive afferents from the caudate nucleus, and (b) those regions of the intermediate layers of the superior colliculus which receive afferents from the latter nigral region, ever resulted in orofacial dyskinesia. These findings support the hypothesis that the anatomically distinct input-output channels of the caudate nucleus are differentially involved in orofacial dyskinesia. The clinical impact of these findings is discussed in view of the L-3,4-dihydroxyphenylalanine-induced tardive dyskinesia in man. In addition, the relevance of the anatomical data is discussed in view of the co-occurrence of Parkinson's Disease and Dementia of Alzheimer type in certain patients. PMID- 2561521 TI - The firing patterns of rat melanotrophs recorded using the patch clamp technique. AB - Cell-attached and whole-cell recordings were made from adult rat melanotrophs maintained in vitro by standard cell culture techniques. In cell-attached recordings the cells showed small biphasic currents which reflected spontaneous cell firing. Single channel currents often had distinct relaxations and depolarizing currents through single channels could trigger the discharge of an action potential in the cell; both observations are consistent with the high input resistance (1-10 G omega) measured in the whole-cell configuration. The discharge of action potentials occurring either spontaneously or by current injection was eliminated by tetrodotoxin or by removing Na from the external medium. A Na-dependent plateau depolarization which activated near the spike threshold was also seen. In cells exposed to tetrodotoxin and K-channel blocking agents it was possible to evoke a long-lasting (up to 20 s) action potential which was enhanced and reduced, respectively, by Ba and Cd and thus appeared to reflect currents through voltage-activated Ca channels. Small amplitude Ca dependent depolarizations could also be evoked at membrane potentials as low as 40 mV. In cell-attached and whole-cell recordings 10 mM Ba caused the discharge of tetrodotoxin-insensitive action potentials prior to a maintained depolarization of the membrane. The low threshold for Ca-dependent depolarizations suggest that Ca influx might occur in these cells even at the resting potential. Additionally, both a Ca current and the current underlying the Na-dependent plateau depolarization may influence the rate of cell firing and in doing so further increase Ca influx through voltage-activated channels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561522 TI - [Acute polyneuropathy in the critical patient. Description of a clinical case]. AB - Polyneuropathy syndromes were described during prolonged sepsis and multiple organ failure. This kind of polyneuropathy should be discriminated from Guillain Barre syndrome. The authors report their experience concerning a clinical case. PMID- 2561523 TI - Crystal deposition diseases. PMID- 2561524 TI - [A seroepidemiologic and virological study of the presence of arboviruses in Moldavia in 1961-1982]. AB - In Romania, St. Draganescu et al., Miszkolczy et al., based on clinical and epidemiological observations, affirmed the possible presence of "tick-borne encephalitis (TBE)". However, critical retrospective evaluation of certain others' papers published through 1958-1971 reveal some uncommon features. By way of example: a very high incidence of CF- and HI-antibodies in acute and chronic CNS illnesses as well as in general population; replication in nucleus and maturation in mitochondria of a strain of "TBE virus" isolated from a patient CSF; isolation of "group B arbovirus" from Ixodes persulcatus on Romanian territory. As known, this tick was not identified in Romania, "no tick-borne Flavivirus isolated outside the U.S.S.R. proved to be a strain of RSSE virus" and "no arbovirus (Alpha- and Flavivirus or Bunyavirus) was ever detected in nucleus". Based on the HI reaction with West Nile (WN) virus, strain Egypt 101, human, domestic and wild vertebrates sera, treated with acetone for removal of nonspecific inhibitors, were tested as part of a control study, in Moldavia in 1961. WN virus was selected for its capacity to crossreact in the HI test with many members of the family Flaviviridae, genus Flavivirus (formerly Group B arboviruses), including the agents of Central European encephalitis. In order to avoid the interference of nonspecific inhibitors, a new single radial haemolysis test for the assay of antibodies to haemagglutinating arboviruses was calibrated in 1979. The seroepidemiological study performed on all 19.853 sera (10.698 human, 6.452 domestic mammals, 2.477 wild small rodents and 226 migrating aquatic birds) pointed out a very low circulation of Flaviviruses in the investigated area. The attempts to isolate such viruses from Ixodes ricinus on suckling mice yielded a single strain of Kemerovo (genus Orbivirus) virus. The need of extending the study to other viruses transmitted by arthropods and rodents, and mainly on the haemorrhagic fever with renal syndrome, an illness already clinically identified in Moldavia, is emphasized. Final consideration of viability (specificity, sensitivity, accuracy and precision) of the tests used and the need for continuous quality control is also presented. PMID- 2561525 TI - [Clinico-epidemiologic and laboratory studies of patients with primary hepatocellular carcinoma]. AB - The investigations carried out in 92 patients with confirmed diagnosis of primitive hepatocellular carcinoma (HCC) have demonstrated that in almost half of them hepatitis was present in their antecedents, indicating the intervention of hepatitis virus in the etiology of this disease. In this context, the primary prophylaxis of primitive HCC is mainly based on the prophylaxis and control of viral hepatitis. As the interval detection-death was in most cases very short (1 2 month), the secondary prophylaxis consisting in performing more of the investigations recommended for the early diagnosis of primitive HCC is essential. PMID- 2561526 TI - Evidence for the presence of disulfide bridges in opioid receptors essential for ligand binding. Possible role in receptor activation. AB - The mobility of purified mu opioid binding protein in SDS-polyacrylamide gek electrophoresis is sensitive to the presence of reducing agents. In the presence of increasing concentrations of DTT the apparent molecular weight increases in a stepwise fashion from 53 kDa to 65 kDa. This reduction in mobility is attributed to the successive breakage of disulfide bridges, resulting in an increasingly asymmetric molecule. Treatment of cell membranes from various brain areas with reducing agents, such as DTT, produced a concentration-dependent inhibition of opioid binding. Sensitivity to DTT inhibition varied between receptor types, mu greater than delta much greater than kappa. For mu receptors, agonist binding was considerably more sensitive to DTT than antagonist binding. Inhibition by DTT is readily reversible and is unaffected by Na+ and/or Mg2+ ions. Reversibility may be partially prevented by the inclusion of a low concentration of a reducing reagent such as glutathione which does not inhibit binding but blocks reformation of disulfide bonds. Scatchard analysis of saturation data shows that DTT causes a pronounced decrease in binding affinity with little effect on receptor number. It is suggested that disulfide bonds are essential for ligand binding and that cleavage of one or more of these bonds may play a role in opioid receptor activation by agonists. PMID- 2561527 TI - Application of a degenerate consensus sequence to quantify recognition sites by vertebrate DNA topoisomerase II. AB - A consensus sequence has been derived for vertebrate topoisomerase II cleavage of DNA (Spitzner, J. R. and Muller, M. T. (1988) Nucleic Acid. Res. 16, 5533-5556). An independent sample of 65 topoisomerase II sites (obtained in the absence of topoisomerase II inhibitors) was analyzed and found to match the consensus sequence as well as enzyme sites determined in the presence of the anti-tumor drug 4'-(9-acridinyl-amino)-methanesulfon-m-anisidide (m-AMSA). As originally described, conventional application of the consensus sequence afforded accuracy in the prediction of the locations but not the frequencies of topoisomerase II cleavages. In the present report, we describe a new method which quantitatively discriminates sites from nonsites, called the 'matrix mean' method (the mean match of a site to the matrix of base proportions from the original consensus sequence derivation). Furthermore, we derived a second method, called the 'unique score' model, which predicts frequency of topoisomerase II activity at a cleavage site. In the unique score method both DNA strands of a site are examined to determine the total number of the consensus positions that match on at least one strand of a potential site. From the new data base of 65 topoisomerase II sites, cleavages were scored for relative cleavage strength. Linear regression analysis showed a significant (p less than 0.01) correlation between the unique score and cleavage strength. The study was extended to show that the unique score model accurately and quantitatively predicts topoisomerase II sites either in the absence or presence of m-AMSA using the same consensus sequence. PMID- 2561529 TI - Model-building of Fnr and FixK DNA-binding domains suggests a basis for specific DNA recognition. AB - The DNA-binding C-terminal domains of the regulatory proteins Fnr from Escherichia coli and FixK from Rhizobium meliloti have been modelled on the basis of their homologies to the CAP protein from E. coli. Residues Glu181, Thr182 and Arg185 of CAP, which are exposed residues of the DNA-recognition helix alpha F, are conserved in Fnr and FixK. However, Arg180 and Gly184 are substituted by Val and Ser respectively in Fnr. We propose that this valine makes a Van der Waals' contact with the first thymine in the Fnr consensus TTGA-N6-TCAA, and that the serine contributes to the binding by displacing a thymine-bound water molecule. The corresponding residues in FixK, Ile and Ser allow the same interactions with a thymine. Therefore we predict that FixK may recognize the same sites as Fnr. This is supported experimentally by showing that Fnr can substitute for FixK in activating the fixN gene in E. coli. PMID- 2561530 TI - Binding characteristics of a series of dimeric tripeptide enkephalins for delta opiate receptors in rat brain and NG108-15 cells. AB - The N-terminal tripeptide enkephalin analogue, Tyr-D-Ala-Gly, was dimerized at the C-terminus systematically with a series of alpha,omega-diaminoalkanes, NH2 (CH2)n-NH2 (n = 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, and 22). The binding affinities of dimers for delta opiate receptors in rat brain were evaluated and compared with those for delta receptors in NG108-15 cells. Although the monomeric tripeptide amide was almost inactive, dimers showed a dramatic increase in binding affinity (8-900 times). The enhancement of affinity was apparently related to the number of methylene chains in the crosslinking spacer moiety, and it was maximal at n = 14-18 in the rat brain. In NG cells the activity increased progressively from n = 2 to n = 22 without reaching any apparent peak. These results suggest that delta receptors in rat brain and NG cells may have slight structural differences. PMID- 2561528 TI - Molecular recognition between oligopeptides and nucleic acids. Specificity of binding of a monocationic bis-furan lexitropsin to DNA deduced from footprinting and 1H NMR studies. AB - MPE-Fe(EDTA) footprinting of a novel monocationic bis-furan lexitropsin 6 on a HindIII/EcoRI restriction fragment of pBR322 DNA revealed a series of four-base binding sites (all 5'----3') of (primary) TGTA, TGAA, AAAT, ACAA, TTAT, and (secondary) CTAA, TCGT, TGTA, GTCA, and GGTT. Thus 6 can accept a GC pair at positions 1, 2 or 3 of the binding site with a strict 3' (4 position) AT requirement. Marked enhancement of cleavage, particularly at GC rich sequences, is observed at regions flanking or even up to 18 base pairs remote from a given binding site. The non-exchangeable and imino 1H NMR resonances of the 1:1 complex and d-[CATGGCCATG]2 were assigned using a combination of NOE differences, NOESY and COSY techniques. 1H NMR studies (ligand induced chemical shifts and NOE differences) of Lexitropsin 6 with d-[CATGGCCATG]2 show unambiguously the location and orientation of the N to C termini of 6 on the sequence 5'-G5C6C7A8 3', with the C terminus oriented to A8. This orientation of 6 in the minor groove of 5'-GCCA is confirmed by an NOE observed between H1 2a of 6 and AH8(8). This preference for binding of 6 to the sequence 5'-GCCA when challenged with d [CATGGCCATG]2 is in accord with the conclusions of the footprinting experiments wherein GC base pairs can be accepted in the first three positions and with a strict 3' terminus AT reading requirement. Collectively the data support the inference of a GC recognizing capacity for a 2,5-substituted furan moiety within a lexitropsin. The 1H NMR data indicate that the decadeoxyribonucleotide duplex exists in the B conformation in both the 1:1 complex and the free form. The apparent binding constant of 6 to calf thymus DNA is 1.68 X 10(5) M-1 whereas netropsin under similar conditions gives a value of 1.85 X 10(7) M-1. This suggests that if advantage is to be taken of the GC recognizing property of a 2,5 substituted furan in longer lexitropsins it should be flanked by more strongly bound moieties. PMID- 2561531 TI - [Cholecalciferol Reference Standard (Control 871) of National Institute of Hygienic Sciences]. AB - Cholecalciferol Reference Standard (Control 871) for the Japanese Pharmacopoeia was prepared. The following analytical data were obtained: melting point, 86.3 degrees C; infrared spectrum, same as the Japanese Pharmacopoeia Standard "Cholecalciferol Standard (Control 851)"; absorbance, E1%1cm (265 nm) = 474.5; optical rotation, [alpha]20D = + 110.0 degrees; thinlayer chromatography, same as the Japanese Pharmacopoeia Standard; highperformance liquid chromatography, contaminants were not detected; assay, 100.7%. On the basis of those results, this material was authorized as the Japanese Pharmacopoeia Standard (control 871). PMID- 2561532 TI - [Correlation of steroid hormone receptors (SR) with clinical and pathological features of human breast cancer]. AB - Paraffin-embedded materials from 104 patients with breast cancer were assayed for the expression of estrogen receptor (ER), progesterone receptor (PR) and androgen receptor (AR) with avidin-biotin complex method. A significant positive correlation was found between SR status and cell differentiation or lymph node status. Tumors with elastosis tend to contain ER, PR and AR more frequently. Patients with positive ER, PR or AR tumors were 1.2 to 2.6 times more likely to survive than those with negative ones. The five-year survival rate increased with the increase of number and amount of positive SR. These results indicate that ER, PR and AR detected by ABC method may be used as biomarkers for tumor differentiation and have prognostic values in breast cancer. PMID- 2561533 TI - [Evaluation of DNA content in gastric dysplasia and carcinoma by image cytometry]. AB - In 30 cases of gastric dysplasia and 10 cases of gastric carcinoma, DNA content was studied by IBAS image analysis system. The mean DNA level increased steadily with the advance of histologic gradation, and the highest DNA content was observed in gastric carcinoma. No case of aneuploidy was found in mild dysplasia. In moderate dysplasia, aneuploid cells were occasionally encountered. Severe dysplasia had a lower percentage (4.48%), and gastric carcinoma was characterized by a high percentage of aneuploid cells (14.54%). PMID- 2561534 TI - [Cytologic diagnosis of salivary gland lesions by fine needle aspiration]. AB - Fine needle aspiration cytology was used in diagnosis of 504 major and minor salivary gland lesions. In 180 cases with satisfactory specimens, cytologic diagnosis was contrasted with pathohistologic findings. There were 150 benign lesions (including 124 tumors and 26 cases of other lesions) and 30 malignant tumors. The total correspondence rate was 94.5%. The rate of misdiagnosis was 5.5%. Cytologic appearance of various salivary gland tumors is described and the significance of distribution of mucus is discussed. The misdiagnosed cases were mainly in the early stage. PMID- 2561536 TI - [Gestational trophoblastic disease. Clinical course and results of treatment]. AB - Out of 472 patients with gestational trophoblastic disease directed to the Institute of Oncology in Warsaw in the period 1.4.1977-1.4.1987, 145 (30.7%) have been qualified for treatment. Lasting cure was obtained in all of 105 patients with the disease restricted to the uterus, in 13 of 14 (92.8%) with metastatic disease with favourable prognosis and in 20 out of 25 (80.0%) with metastatic disease with poor prognosis. In 102 patients treated exclusively chemically the procreative ability has been preserved, 33 of these women gave birth to a total of 44 healthy children. PMID- 2561537 TI - [Role of radiotherapy in the treatment of patients with non-small-cell lung cancer]. AB - Recent opinions concerning the role of radiotherapy in the treatment of patients with non-small-cell lung cancer are presented, particular attention being called to the problem of independently applied radical irradiation, adjuvant postoperative irradiation and palliative irradiation. PMID- 2561535 TI - A reinterpretation of Na channel gating and permeation in terms of a phase transition between a transmembrane S4 alpha-helix and a channel-helix. A theoretical study. AB - A functional model for the S4/IV alpha-helix of the action potential sodium channel is described by means of a thermodynamic approach. The model is based on a phase transition between the alpha-helix and an ion conducting channel-helix which is similar to the well established helix-coil transition in solution. The right hand channel-helix is a peptide chain with an alternating sequence of torsional angles (phi1, psi1) = (87 degrees, 315 degrees) and (phi2, psi2) = (22 degrees, 107 degrees) which yields a helix of 13.5 A per turn. The axial dipole moments of the peptide bonds of this chain of L-amino acids nearly cancel each other out in a similar way to those in the gramicidin A channel, which is formed by alternating D- and L-amino acids. The helix, which does not contain any H bonds, is stabilized by a helical file of water molecules which includes the permeating ion(s). This file turns around the channel-helix to form a relatively stable "double helix" structure which corresponds to the open channel. Since every third side chain in the S4/IV helix carries a positive charge their environments must be polarized. These polarized regions form a left hand screening-helix around the alpha-helix with 27 A per turn. If all H-bonds of the alpha-helix are broken and the internal alpha-carbon atom is considered as fixed, the outer ten residues leave the membrane while the internal ten residues form the channel-helix. In this configuration every positively charged side chain matches nearly exactly every second polarized region of the screening-helix leaving the three regions in-between exposed to the water file containing the ion(s). This further stabilizes the channel and agrees nicely with the idea of cationic selectivity. An analysis of the energetics of the alpha-helix-channel helix transition showed that the voltage-independent part of the free energy per helix residue could well be close to 0 kcal/mol and thus be in the range where a transition could occur. Two voltage-dependent contributions were included: the break down of the considerable dipole of the alpha-helix and the outward shift of the positive charges of the side chains upon channel-helix formation. Taking into account the fact that the formation of an alpha-helix is a highly cooperative process the degree of voltage dependence of the probability of formation of a channel-helix proved to be in the same range as experimental values for the open probability of modified Na channels whose inactivation had been removed. With regard to gating currents, the model predicts that 2.74 positive charges are moved in an outward direction. Consequences of the model for other experimental findings are discussed. PMID- 2561538 TI - Effect of ultraviolet radiation on Ia expression by keratinocytes. AB - Many skin diseases, such as graft-versus-host disease (GVHD), are marked by lymphocyte infiltrates in the skin. Severity of these diseases is often correlated with the induced expression of class II antigens (human, HLA-DR,; murine, Ia) by the keratinocytes. This suggests that HLA-DR-expressing keratinocytes may be involved in the pathogenesis of these diseases. Since some of these diseases are effectively treated with ultraviolet radiation (UVR), this study was conducted to determine whether UVR alters the keratinocyte expression of class II antigens. To test this hypothesis, 2 models of experimentally induced keratinocyte Ia expression were employed. First, athymic nude mice with one ear protected by electrical tape were exposed to UVR (450 J/m2/day on 4 consecutive days). They were then given an i.v. injection of normal mouse serum (NMS) to induce keratinocyte Ia expression. Keratinocytes in the UVR-exposed skin of these animals were not induced to express Ia; however, Ia-expressing keratinocytes were observed in the epidermis of shielded skin sites. Likewise, it was determined that UVR was capable of downregulating keratinocyte expression of Ia when administered to nude mice 7 d after receiving an injection of NMS. Second, employing a clinically relevant model, we found that Ia expression by keratinocytes in mice undergoing experimentally induced GVHD was abrogated by UVR treatment. This appeared to be a direct effect of the UVR, since keratinocytes in shielded skin sites and mucosal cells in the intestinal epithelium of animals with GVHD were shown to express Ia. These data provide compelling evidence for our hypothesis that decreased HLA-DR expression by keratinocytes in diseased skin treated with UVR is a mechanism by which UVR exerts its therapeutic effect. PMID- 2561540 TI - The role of intracellular proteinases in human neutrophil activation. AB - In addition to other proteinases human neutrophils contain two non granular neutral endopeptidases: a serine proteinase and a cysteine Ca2+ dependent proteinase named calpain. Serine proteinase localized in association with the cytoskeleton-membrane proteins, apparently exerts a dual role: it is partially released into the medium during neutrophil stimulation by phorbol myristate acetate (PMA), presumably acting as one of the cytotoxic factors; and in its intracellular localization is presumably involved in the process of down regulation of native protein kinase C (PKC). Calpain, predominantly present in resting conditions in an inactive form, becomes activated in the course of neutrophil stimulation and appears to be involved both in the down regulation of native PKC as well as in the formation of a proteinase-activated kinase form, presumably derived from PKC and defined as PKC-M. Calpain once activated appears to be also involved in cytoskeleton rearrangement, through proteolytic degradation specifically oriented by substrate phosphorylation. Activation, down regulation of PKC, formation of the proteinase-activated kinase, as well as proteolytic processing of cytoskeleton have been demonstrated to be correlated to those biochemical responses which characterize neutrophil activation. PMID- 2561539 TI - The non-lysosomal, calcium-dependent proteolytic system of mammalian cells. AB - The intracellular calcium-dependent proteases (calpains) and their endogenous protein inhibitor (calpastatin) are present in many different mammalian cells. There is emerging evidence for their importance in the turnover of membrane associated proteins. Accordingly, it is important to understand how these proteinases and their inhibitor interact within cells, in particular at membranes. Bovine myocardial calpastatin appears to be associated in part with intracellular membranes, where it may effectively block the activity of calpain II on membrane-associated proteins. Immuno-electron microscopic studies suggest that canine myocardial calpain and calpastatin are associated with a number of membranous organelles. During canine myocardial autolysis, the amount of calpain at various organelles decreased, but the amount of calpastatin decreased to an even greater extent. Thus there may be a high calpain to calpastatin balance during heart ischemia at these sites. Calpain II aggregation may contribute to localization of the proteinase at sites of high calcium concentration within cells. A model is presented for interaction of calpain II and calpastatin at cellular membranes in the presence of calcium. PMID- 2561541 TI - ATP-dependent mechanisms for protein degradation in mammalian cells. AB - The biochemical basis for ATP-dependent protein degradation observed in intact cells has been studied in cell-free extracts of baby hamster kidney fibroblasts. ATP plays at least two distinct roles in proteolysis. First, ATP is required for the covalent conjugation of ubiquitin to protein substrates. This modification markedly enhances the rates of degradation of some, but not all, proteins. Second, ATP appears to stimulate the activity of a protease capable of degrading both ubiquitinated and non-ubiquitinated proteins. This protease has several biochemical and catalytic features that resemble those of the previously described high molecular weight protease, macropain. Furthermore, antibodies against highly purified human erythrocyte macropain inhibit both ubiquitin dependent and ubiquitin-independent pathways of ATP-dependent proteolysis. Such results provide evidence for an important role for macropain in ATP-dependent proteolysis in mammalian cells. PMID- 2561542 TI - Protease/inhibitor mechanisms involved in ATP-dependent proteolysis. AB - ATP-dependent proteolytic activities constitute a family of distinct enzymes and enzyme complexes which function in the selective degradation of proteins in the cytosol and possibly other non-lysosomal cellular compartments. In reticulocytes as well as other eukaryotic cells, conjugation of the polypeptide ubiquitin to proteins, an ATP-requiring process, is important for proteolysis. However, the popular view that ubiquitin functions by tagging substrates for recognition by conjugate-specific proteases should be viewed with caution. Evidence is reviewed implicating an ATP-independent high molecular weight protease (HMP) which can degrade non-ubiquitinated proteins in the ATP-dependent pathway. This protease consists of several related subunits based upon monoclonal antibody studies. HMP by itself, can account for effects of substrate amino group blockage on ATP dependent proteolysis. Endogenous inhibitors (40 kDa and 50 kDa) have been isolated and shown to block HMP non-competitively. Unlike the soluble system in reticulocytes, the major ATP-dependent activity in mouse erythroleukemia cells is particulate in aqueous media but soluble in the presence of sucrose. Despite such differences, evidence suggests analogous inhibitory components in this system. The possibility that protease/inhibitor interaction is regulated by ATP and ubiquitin in different systems is discussed. PMID- 2561543 TI - The ubiquitin-dependent proteolytic pathway: specificity of recognition of the proteolytic substrates. AB - Degradation of intracellular proteins via the ubiquitin system involves several steps. Initially, ubiquitin is covalently linked to the protein substrate in an ATP-dependent reaction. Following ubiquitin conjugation, the protein is selectively degraded with the release of free and reutilizable ubiquitin. Ubiquitin modification of a variety of protein targets within the cell appears to be important in a number of basic cellular functions. For example, modification of core nucleosomal histones may regulate gene expression at the level of chromatin structure. Ubiquitin attachment to cell surface proteins can play a role in the processes of cell-cell interaction and adhesion. Conjugation of ubiquitin to other, yet to be identified protein(s), is probably involved in the progression of cells from one stage to another in the cell cycle. Despite the considerable progress that has been made in elucidating the mode of action and roles of the ubiquitin pathway major problems remain unsolved. One problem of central importance is the issue of what determines the specificity of the ubiquitin ligation system for commitment of certain proteins to degradation. While a free alpha-amino group is one structural feature of the substrate recognized by the ligation system, it is certainly not the only one, and possibly not the predominant recognition marker. The scope of this review is to discuss recent developments in our understanding of the specificity and selection of substrates for conjugation and subsequent degradation via the ubiquitin system. PMID- 2561544 TI - PEST regions, proteolysis and cell cycle progression. AB - How eucaryotic cells control cell division is both a major intellectual challenge and a problem of considerable medical importance. A decade of studies on oncogenes has made it abundantly clear that protein kinases play a central role in regulating mitosis. In this essay, I review a number of recent discoveries providing evidence that selective proteolysis, like phosphorylation, is an important mechanism for ordering metabolic events within the cell cycle. PMID- 2561545 TI - Ubiquitin-protein conjugates: clinical and experimental findings. AB - Ubiquitin has been extensively studied as a protein which is a cofactor in extralysosomal protein degradation, particularly in reticulocyte lysates. Ubiquitin is also found conjugated to nuclear histones and surface receptors in somatic cells. Until recently the occurrence of stable cellular ubiquitin-protein conjugates in physiological and pathological states had not been considered and studied. Recently we have shown that ubiquitin-protein conjugate immunoreactivity is a clinical feature in several ostensibly unrelated chronic human degenerative diseases as well as in some viral diseases. The consistent observation is the occurrence of intracellular extralysosomal inclusions containing intermediate filaments and ubiquitin conjugates as determined by immunohistochemical methods. These diseases are therefore part of a family of intermediate filament-ubiquitin diseases. The involvement of intermediate filaments with ubiquitin, a protein of known significance in protein degradation, ties in with separate evidence for a close role between intermediate filaments and protein degradation. We have previously shown that intermediate filaments may be involved in protein sequestration for degradation in the lysosomal system. Clinical immunohistochemical observations suggest that elements of the lysosomal degradation system and the ubiquitin-dependent extralysosomal system are involved in the molecular pathogenesis of some diseases. To underpin these clinical observations, we have recently shown that ubiquitin-protein conjugates accumulate in lysosome-related multivesicular bodies in cells in which lysosomal degradation is impaired. This phenomenon may result from increased ubiquitin protein conjugate formation in cells with a compromised lysosomal system followed by chance uptake into multivesicular bodies. Alternatively, ubiquitination may normally serve as a signal for protein uptake into the lysosomal system, ubiquitinated protein-conjugates may therefore accumulate in cells with a functionally impaired lysosomal system. PMID- 2561546 TI - [Chronic hepatitis B: anatomo-clinical, serologic and developmental aspects]. AB - Few data on chronic hepatitis B (CHB) have been published in our country, despite the fact that it is responsible for more than 50% of all types of chronic hepatitis. From 1968 to 1988, 164 patients were attended with the diagnosis of CHB, from whom 136 (82.9%) were male. Only 11 (8.1%) admitted homosexual behavior. Twenty six out of 39 (66.7%) health professionals were medical doctors; among them 12 (46.2%) were surgeons. The mode of transmission was unknown in 55% of the cases, but vertical and sexual transmissions were also frequent. Commercial gammaglobulin, used with prophylactic purpose, was probably responsible for eight cases between 1972 and 1975. The most frequent forms of CH were chronic active hepatitis (CAH) and liver cirrhosis (LC): 72 or 43.9% and 53 or 32.3%, respectively. The predominance of HBeAg (66.4%) was observed in all forms of CHB. Repeated biopsies showed that chronic lobular hepatitis (CLH) and chronic persistent hepatitis (CPH) may occasionally progress to CAH. This form may persist as such for some years or progress to cirrhosis. In a few cases the evolution to CPH was observed. In the long term follow-up of our patients, the appearance of hepatocellular carcinoma was observed in 8 (4.9%). PMID- 2561547 TI - Molecular epidemiology of human rotavirus infection in Coro, Venezuela. AB - From December 1984 to December 1987, rotaviruses were detected in 115 (24%) of 470 hospitalized children, by electrophoretic analysis of viral RNA. Three peaks of increased incidence were observed, coincident with the cooler months of each year. Rotavirus was found in 31% of the children with diarrhea and in 47% of those who were severely dehydrated. Vomiting was significantly associated with rotavirus shedding in diarrheic children. Eleven electropherotypes were identified, showing a sequential pattern of appearance. Strains exhibiting a "short" pattern were present in 10% of the positive cases. The rate of infection increased to 71% in march 1987, when a mixed infection with two genotypically different strains was found and when electropherotypes with an extra RNA segment appeared. PMID- 2561548 TI - Agonist derived molecular probes for A2 adenosine receptors. AB - The adenosine agonist 2-(4-(2-carboxyethyl)phenylethylamino)-5'-N- ethylcarboxamidoadenos ine (CGS21680) was recently reported to be selective for the A2 adenosine receptor subtype, which mediates its hypotensive action. To investigate structure/activity relationships at a distal site, CGS21680 was derivatized using a functionalized congener approach. The carboxylic group of CGS21680 has been esterified to form a methyl ester, which was then treated with ethylenediamine to produce an amine congener. The amine congener was an intermediate for acylation reactions, in which the reactive acyl species contained a reported group, or the precursor for such. For radioiodination, derivatives of p-hydroxyphenylpropionic, 2-thiophenylacetic, and p aminophenylacetic acids were prepared. The latter derivative (PAPA-APEC) was iodinated electrophilically using [125I]iodide resulting in a radioligand which was used for studies of competition of binding to striatal A2 adenosine receptors in bovine brain. A biotin conjugate and an aryl sulfonate were at least 350-fold selective for A2 receptors. For spectroscopic detection, a derivative of the stable free radical tetramethyl-1-piperidinyloxy (TEMPO) was prepared. For irreversible inhibition of receptors, meta- and para-phenylenediisothiocyanate groups were incorporated in the analogs. We have demonstrated that binding at A2 receptors is relatively insensitive to distal structural changes at the 2 position, and we report high affinity molecular probes for receptor characterization by radioactive, spectroscopic and affinity labelling methodology. PMID- 2561549 TI - [Cytomegalovirus infection in neonate--report of two cases]. AB - The cytomegalovirus (CMV) is an abiquitous agent that infects almost all human beings at some time during their lives. In developing area of the world, 90% or more of the population is infected during childhood. However, in developed countries the infection is acquired at a lower rate. In neonates, CMV infection can be divided into congenital and perinatal infection. Congenital CMV infection is the result of transplacental transmission, CMV can be transmitted to the fetus following reactivation as well as primary infection during pregnancy. The incidence of congenital infection is 0.2% to 2.2%. Symptomatic congenital CMV infection is more likely to be the result of primary as opposed to reactive CMV infection during pregnancy. The clinical manifestations of symptomatic CMV infection are hepatosplenomegaly, microcephaly, jaundice, petechiae, small for gestation age, periventricular calcification and chorioretinitis. Mortality may be as high as 30% among the most severely affected infants. In the survivors, about 90% will develop mild to severe handicaps. Perinatal CMV infection can be acquired from exposure to virus in the maternal genital tract at delivery, breast milk, or through blood transfusion. In premature infants who require prolonged and intensive medical care, blood transfusions are an important iatrogenic cause of CMV infection. Transfusion-acquired perinatal CMV infection can cause significant morbidity and mortality, particularly in premature infants with a birth weight of less than 1250 gm born to CMV-seronegative mothers. So CMV acquisition can be prevented either by providing these infants blood products from seronegative donors or by using frozen deglycerolized red blood cells. Two cases of neonatal CMV infection are reported, one with congenital infection, the other with perinatal infection. PMID- 2561550 TI - [Collagen and fibrin as biologic binders for granular hydroxylapatite]. AB - In an animal experiment the use of the biologic binders collagen and fibrin for the temporary stabilization of the shape of granular hydroxyl apatite grafts has been tested. A simple way of producing a hydroxyl apatite fibrin paste involves the use of a thrombin solution diluted to 1 i.u./ml. The fibrin-bound polygonal granules (Interpore 200) studied did not grow denser during the interposition of fibrous tissue, while the molded specimens of collagen-bound granules (Alveoform) underwent condensation involving a 30% decrease in intergranular distance. PMID- 2561551 TI - [Alveolar ridge augmentation using moldable implants of granular hydroxylapatite and fibrin adhesive]. AB - First results in the use of a hydroxyl apatite fibrin paste for alveolar ridge augmentation are reported. In the production of the moldable implants a fibrin mixture containing 1 i.u. Thrombin/ml is used as an absorbable biologic binder for the granules. The individual shape and situation of the graft is maintained throughout the resorption of the fibrin adhesive and the spread of connective tissue into the intergranular spaces, without any additional means of fixation required. Even under functional load, which starts after 6 weeks, the intergranular distance and the shape of the implants remains unchanged. PMID- 2561552 TI - [Primary sulcoplasty with hydroxylapatite augmentation for extreme ridge resorption]. AB - Hydroxylapatite is commonly used for augmentation of the atrophic alveolar ridge. A new surgical technique is described for HA-augmentation and simultaneous vestibuloplasty on the severely atrophic jaw. The open surgical technique includes a lingually based mucoperiostal flap, the implantation of Hydroxylapatite particles in a vicryl mesh and the use of a positioning splint. First results show a sufficient depth of the vestibular sulcus and only few complications (circumscripted mucosal erosions). No lateral or lingual dislocation of the implanted HA-particles and no permanent mental nerve anaesthesia were observed. The prosthodontic results show increased stability and retention of the dentures following the one stage surgery. PMID- 2561553 TI - Serum alpha-1-antitrypsin and alpha-1-antichymotrypsin in the diagnosis of primary hepatocellular carcinoma. AB - Both alpha-1-antitrypsin (AAT) and alpha-1-antichymotrypsin (AAC) in serum were investigated in various liver diseases. Serum levels of AAT and AAC in normal controls were 293.3 +/- 37.9mg/dl and 119.6 +/- 18.9U/ml, respectively. Both AAT and AAC were significantly higher in 75 patients with primary hepatocellular carcinoma (PHC) than in patients with acute viral hepatitis, chronic hepatitis, liver cirrhosis as well as normal controls. In a cut-off value of more than 400 mg/dl AAT showed a sensitivity rate (SS) of 74.7%, specificity (SP) of 97.3%, positive predictive value (PV+) of 86.2%, negative predictive value (PV-) of 95.5% and a total accuracy of 93.3% for diagnosing PHC. In a cut-off value of more than 157 U/ml, the corresponding figures for AAC were 68.0%, 96.7%, 86.4%, 90.6% and 90.0%. If the increased levels in one or both tests are positive, the combined positive rate is 80.0% in PHC. In 11 PHC cases with serum alpha fetoprotein (AFP) less than 50 ng/ml, the positive rate of AAT and AAC were 72.7% and 81.8%, respectively, compared to corresponding figures of 71.7%, and 65.2% in 46 cases of PHC with more than 50 ng/ml of AFP. It shows that combined assay of both serum AAT and AAC has a complemental value in the diagnosis of PHC, especially in patients without increased AFP. PMID- 2561554 TI - A study of molecular epidemiology of adenovirus of types 3 and 7 on infant pneumonia in northern China. AB - This paper analysed 102 strains of adenovirus types 3(Ad3) and 7(Ad7) causing infant pneumonia from 1976 to 1988 in northern China. Two genotypes of Ad7, 7b and 7d, were identified by using restriction endonucleases, BamHI, BcLI, BgLI, SmaI, XbaI and HindIII. 3 genotypes of Ad3, 3I, 3II and 3III, were identified by using BgLII and BamHI. Of 56 Ad7 strains, 34 were 7b (76.8%) spread over last 10 years; 13 7d(23.2%) occurred from 1982, together with 7b. Of 46 Ad3 strains, 42 3I(91.3%) spread over the past 12 years. 3II and 3III scattered all over these years. Ad3I and Ad7b were the dominant genotypes. The results indicated that Ad7d tended to increase with time from 1982. It is possible that Ad7d will become dominant genotype and replace Ad7b. PMID- 2561556 TI - Oxygen free radical in human osteoarthritis. AB - The xanthine oxidase catalysed release of superoxide free radicals (O2-) from endogenous hypoxanthine was determined in homogenates of synovium obtained from three groups of patients, those undergoing primary or revision total hip replacement (THR) for osteoarthritis and those undergoing arthroscopy of the knee for semilunar disc injuries. The concentrations of hypoxanthine in homogenates obtained during THR were found to be significantly higher than those in the group with semilunar disc injuries. The results suggest that there is a greater predisposition to free radical release and tissue damage in osteoarthritis. PMID- 2561555 TI - Effects of aminoglutethimide on hypothalamic-pituitary-adrenal functions. AB - This study was performed on four groups of subjects, including 10 patients with Cushing's disease, 10 patients with simple obesity, 8 patients with hypopituitarism and 13 normal subjects. The study was conducted by measuring the sequential changes of plasma ACTH, serum cortisol, 24-h UFC, 24-h 17 KS and 24-h 17 KGS following aminoglutethimide (AG) administration. The results suggest that normal subjects showed sequential changes of hypothalamic-pituitary-adrenal hormone concentrations with normal feedback regulation of the axis following AG administration. Patients with Cushing's disease had obvious autonomy in the production of ACTH from the pituitary. Patients with simple obesity might display abnormality to some degree in the production from the pituitary. Patients with hypopituitarism lost the capacity of ACTH production in various degrees because of pituitary lesions. PMID- 2561557 TI - Effects of sonication and growth temperature on the cytochrome oxidase activity of Bacillus species. AB - The cytochrome oxidase activity of Bacillus strains was analyzed quantitatively by colorimetric N, N, N', N'-tetramethyl-p-phenylene-diamine (TMPD) oxidase assay. Twenty six type strains were readily grouped, the oxidase positive, the oxidase negative, and the oxidase indeterminate groups. TMPD-dependent oxidase activities for whole cells and sonicated cells were compared. Spectral absorbance analyses of membrane fractions showed that all 26 Bacillus type strains contained the o-type cytochrome oxidase; 19 strains contained cytochrome a + a3 and among which 5 strains contained cytochromed. All of the 10 thermophilic Bacillus strains, which were predominantly oxidase positive, exhibited maximum oxidase activity when grown at optimum temperature (65 degrees C). The other three "caldo active" thermophiles exhibited maximum oxidase activities when grown at their optimum temperatures, 70 degrees C to 80 degrees C. Mesophilic B. cereus and B. subtilis exhibited maximum oxidase activity in cells grown at 42 degrees C and 55 degrees C, respectively. In no case did growth temperature induce oxidase negative strains to exhibit an oxidase positive reaction, and vice versa. PMID- 2561558 TI - The subtypes, distribution and location of human papillomavirus DNA in genital warts and genital Bowen's disease--a study using in situ DNA.DNA hybridization. AB - The aim of this study is to investigate the difference in subtypes and distribution of human papillomavirus (HPV) between the benign pathological condition- Genital wart (condyloma acuminatum, verruca- like lesion and papular lesion) and genital Bowen's disease (Bowenoid papulosis, Mollucum contagiosum with Bowenoid papulosis and Condyloma acuminatum with verruca- like lesion and pathological bowenoid change) of genitalia by in situ DNA.DNA hybridization. In situ DNA.DNA hybridization was performed by hybridizing the RNAse treated and denatured frozen skin section with 3H-labeled HPV 6, 11, 16 and 18-DNAs probes and then developed autoradiogram on a glass slide. The results reveal that: (1) The benign cases are strongly associated with HPV 6/11, while the bowenoid cases are associated with HPV 16/18, indicating 16 positive in 24 cases and 3 positive in 5 cases respectively; (2) Four of sixteen benign cases were doubly infected with HPV 6/11 and 16/18; and 2 of 2 bowenoid cases were infected with HPV 16/18 only (3) In benign cases, HPV distributes through the upper third of epidermis and in bowenoid cases, HPV scatters throughout the whole epidermis including parabasal layers. These findings may indicate that the subtypes of HPV, the interaction of weak oncogenic virus (HPV 6/11) and strong oncogenic virus (HPV 16/18) and the presence of HPV on proliferating cells (parabasal layers) all play a part in oncogenicity. The detectability of HPV 6/11 DNA in both the nucleus and cytoplasm indicate that: 1). In situ DNA.DNA hybridization is a more sensitive method than the immunological detection of capsid antigen. 2). In addition to the nucleus, the cytoplasm is a site through which HPV 6/11 virus should pass during their life cycle. Using PEG hybridization mixture, it was revealed that 6 of the 6 HPV 6/11 positive condyloma acuminata harbored HPV 6. This result indicates that the subtype of HPV determine the gross morphology of skin lesion. PMID- 2561559 TI - The in vitro effects of high-Tc-superconducting particles (YBa2Cu3O6-7) and quartz (SiO2) on bovine alveolar macrophages. AB - The cytotoxic effects of YBa2Cu3O6--7-particles on cultured bovine alveolar macrophages were investigated by monitoring the release of lactate dehydrogenase and N-acetylglucosaminidase into the culture medium and testing the viability of the cells. After 20 hours of incubation the effects of YBa2Cu3O6--7-particles were very similar to those of analogous mass concentrations of DQ 12 quartz. PMID- 2561560 TI - Infection and growth of herpes simplex virus (HSV) in rabbit's corneal culture cells. AB - With the view of making quantitative examination of the susceptibility of corneal epithelial cell and stromal cell to herpes simplex virus (HSV) infection, 6 HSV strains (4 of HSV type I, 2 of HSV type II) were inoculated in cultured rabbit's corneal cells (epithelial and stromal cells), and the following results were obtained: 1. When stock HSV was titrated with monolayer culture of each epithelial or stromal cells, strong CPE appeared within 24 hr in epithelial cell and the TCID50 titer of HSV was read as 10(7.5)-10(8.7)/ml at 120 hr. On the other hand, in stromal cell the TCID50 titer showed 10(6.4)-10(6.8)/ml at 72 hr and did not increase thereafter. 2. In terms of the growth curve for HSV type I, the eclipse period was about 4 hr for epithelial cell compared with about 4 hr for stromal cell. At 24 hr, postinfection of standard strain, about 10(7) TCID50/ml of virus was obtained from both epithelial and stromal cell culture. In the case of clinical isolate infection, however, about 10(7) TCID50 was obtained from epithelial cells and 1 in 10th lower titer of virus was obtained from stromal cells. The above results seemed to be of importance in the consideration of different onset mechanisms of epithelial type corneal herpes and stromal ones. PMID- 2561561 TI - Partial nephrectomy for tumour. AB - Thirty-three cases of malignant renal tumour removed by partial nephrectomy are discussed. The indications of this conservative therapy as well as the surgical technique, the results obtained, especially survivals beyond 10 years, as well as the period of time after which the neoplastic disease can be considered as cured are presented. PMID- 2561562 TI - DNA topoisomerase: the mechanism of resistance to DNA topoisomerase II inhibitor VP-16. AB - K6-1 and 50B-3 cell lines, resistant to VP-16, a DNA topoisomerase II inhibitor, were established from two different types of cells respectively: human T-cell derived acute lymphoblastic leukemia cell line RPMI8402 and mouse mammary tumor cell line FM3A. IC50 values of K6-1 and 50B-3 cells to VP-16, evaluated by the colony forming ability on methyl cellulose medium, were 11- and 84-fold higher than their sensitive parental cell lines, respectively. Membrane permeability of the drug was not responsible for the resistance in K6-1 and 50B-3 cells. Quantitative analysis of drug-induced DNA cleavage (so called cleavable complex formation) was performed using 32P end-labeled pBR322 restriction fragments. The formation of the topoisomerase II-DNA cleavable complex stimulated by VP-16 in 50B-3 cells was approximately 1/5 compared with that of FM3A wild-type cells. Dot blot analysis of RNA extracted from these cell lines showed that the levels of mRNA for DNA topoisomerase II in 50B-3 cells were markedly decreased and that catalytic activity was reduced to 1/2-1/3 compared with that of parent cells. There was a slight reduction of DNA topoisomerase II mRNA in K6-1 cells. However, DNA topoisomerase II activities were similar in wild-type and K6-1 cells. In addition, 50B-3 cells showed cross resistance to VM-26, m-AMSA and adriamycin, whereas K6-1 cells exhibited increased resistance only to VM-26. These resistant cell lines did not show collateral sensitivity to CPT-11, a DNA topoisomerase I inhibitor. Southern blot analysis of genomic DNA did not show any change in the restriction pattern of the DNA topoisomerase II gene between the parental and their resistant lines. These findings suggest that the reduced levels in DNA topoisomerase II contribute to the drug resistance of 50B-3 cells. PMID- 2561564 TI - [Experimental evaluation of apatite coated implants prepared by the build-up technique]. AB - In this study, apatite coated subperiosteal and endosseous implants were prepared by the build-up technique, that is, dental alloy was cast in the required shape, and a covering of glass was fused onto it. Then hydroxyapatite powder was coated on the surface of the glass coat and hydrostatically pressed to form a compact layer which was then heated. These implant materials were tested by animal experiment to evaluate the applicability for clinical use. The results suggested that endosseous subperiosteal implants are available for maxillary molar parts. PMID- 2561563 TI - [Clinical evaluation of synthetic hydroxyapatite implants for human periodontal osseous defects. 6 cases studied more than 3 years after the surgery]. AB - We applied of granular hydroxyapatite HAP to periodontal bone defects, for the purpose of restoring the periodontal tissue destroyed by periodontal disease, the subjects were 6 patients (6 regions) with chronic marginal periodontitis who had passed more than 3 years since HAP implantation. After routine periodontal initial treatment, a flap operation was performed, and granular HAP, or a combination of granular and block-like HAP, was implanted into the periodontal bone defect. Clinical parameters were probing depth, clinical attachment level, clinical gingival margin, residual HAP volume and X-ray before operation, at 3, 6, 12 months, and 2, 3, and 4, 6 years after the operation. The results revealed that all measured values showed good results for both granular HAP and the combination of granular and block-like HAP after implantation, and that all cases showed stable results from 12 months on. No remarkable difference was noted between the implanted regions. PMID- 2561565 TI - [Effect of various implant materials on cementogenesis]. AB - Various implant materials have been used to stimulate the regeneration of supporting bone lost from periodontal disease. In addition, the histologic features of bone regeneration associated with their implantation have been evaluated. Very little, however, seems to be known about the effect of implant materials on cementum formation. The aim of this study was to determine whether implant materials stimulate the cementogenesis on adjacent planed root surfaces. Twelve monkeys with healthy gingivae were used in this experiment. Following mucoperiosteal flap elevation, "windows" were chiseled in the bone to the proximal root dentin surfaces and adjacent root surfaces were planed. Each of the three implant materials [tricalcium phosphate (TCP), decalcified bone matrix (DBM) and hydroxyapatite (HA)] were then placed in the cuspid and incisor root "windows" before the flap was sutured back into the previous position. Windows with no implantation served as a control. Animals were sacrificed 2, 4 and 8 weeks postoperatively. Biopsy specimens including the tooth and surrounding bone were examined by light and electron microscopy. At 2 weeks, all implant particles were surrounded by fibrous tissue. On the other hand, fibrous tissues filled the control defect. On the planed root surfaces after the implantation of TCP and DBM, furthermore, cementoid tissue appeared. At 4 weeks, a considerable amount of new cementum was deposited on the root surfaces except in the implantation of HA. It was especially pronounced after implantation of TCP and DBM which promoted bone regeneration after resorption. These results suggest that resorbable implant materials such as TCP and DBM not only facilitate the formation of new bone, but also of new cementum. PMID- 2561566 TI - Permeability of apical barriers. AB - The purpose of this work was to measure the permeability (hydraulic conductance) of root canal apical barriers in vitro. In 30 extracted teeth, the pulp tissue was removed and the apical opening standardized and enlarged using endodontic files. One-millimeter thick barriers of either autogenous dentin chips, calcium hydroxide powder, or durapatite particles were placed just inside the apical opening. The prepared teeth were mounted in a special chamber, fluid pressure applied, and the resultant flow measured. The permeability of dentin chip barriers in 10 additional teeth with nonstandardized apical openings was measured also. The standardized canals with the autogenous dentin chip barriers had a mean permeability of 0.0010 microliter/s/psi at 20 psi applied pressure. Teeth with calcium hydroxide barriers averaged 0.0012 microliter/s/psi. The durapatite particle barriers were much more permeable and averaged 0.0220 microliter/s/psi. It was concluded that this technique is useful in evaluating the effectiveness of apical barrier materials. For the materials tested, autogenous dentin chips and calcium hydroxide barriers have significantly lower permeability than durapatite barriers (p = 0.05). PMID- 2561567 TI - [Adenoidcystic carcinoma of the palate in pregnancy: report of a case]. AB - The Patient was a 28-year-old woman with 8th month of her pregnancy. She had noted a swelling of her palate (6-region) about 3 years before. Incisional biopsy was done in 8th month of her pregnancy and histological diagnosis was confirmed; adenoidcystic carcinoma. After normal health baby was born with cesarean section in 34 weeks, partial maxillectomy was performed. She is now free of the disease. The baby is now 1-year-old and enjoying the good health. PMID- 2561568 TI - Thymopentin induces release of ACTH-like immunoreactivity by human lymphocytes. AB - Peripheral mononuclear (PMN) cells are known to produce ACTH-like immunoreactivity (ACTH-LIR) in vitro. Based on these findings the aim of this study was to find out whether thymopentin (the active pentapeptide of the native hormone thymopoietin) may stimulate ACTH-LIR production and release by cultured normal human lymphocytes. Thymopentin at concentration of 1 microgram/ml was capable of inducing ACTH-LIR release by normal human PMN cells (median 22 pg/ml) whereas ACTH-LIR inside cells was lower (median 11 pg/10(7) cells). The chromatographic characterization of the eluted material identified the presence of ACTH immunoreactive peptides with the elution characteristics of the precursors 31 K proopiomelanocortin, 22 K ACTH and 4.5 K ACTH, together with higher molecular weight material (greater than 43 K). These data demonstrate that thymopentin induces ACTH-LIR release by human lymphocytes, thus adding a novel factor to those already reported (corticotrophin releasing factor, lipopolysaccharide, viruses) capable of such function. PMID- 2561569 TI - Axonal dystrophy in the posterior column nuclei of young adult epileptics with chronic phenytoin intoxication. AB - Axonal spheroids in the posterior column nuclei of phenytoin-intoxicated epileptics were classified according to their predominant subcellular components into six types, and their incidences were compared with those in controls. Spheroids from phenytoin-intoxicated epileptics showed significantly higher proportions of the tubulomembranous (TM) and layered membrane loop (LML) types in the gracile nucleus, appearance of the same types in the cuneate nucleus, and a significant decrease of the neurofilamentous (NF) type in both nuclei. The incidences of the complex body (CB) and granular material types and of the homogeneous dense-body (HDB) type, which appeared only in the gracile nucleus, showed no difference between the intoxicated patients and the controls. The NF, CB and HDB types were therefore considered to be nonspecific. It was thought that chronic phenytoin intoxication might induce dystrophic changes, such as those characterized by the presence of the TM and LML types, in the axon terminals of the gracile and cuneate nuclei, possibly due to some abnormalities of the axoplasmic transport system. PMID- 2561570 TI - Fatty acid analysis of galactolipids and ganglioside in the brains of four cases of Nasu-Hakola disease. AB - A study was conducted on the fatty acid composition of cerebroside, sulfatide and ganglioside in the brains of 4 cases of Nasu-Hakola disease. The percentage of short carbon chain (C.16-C.18) nonhydroxy fatty acids of sulfatide in the cortex was much higher than that of the control and long-chain (C.24-) fatty acids showed a lower percentage. Sulfatide in the white matter was of the same tendency. The percentage of C.16:0 palmitic acid of ganglioside in the cortex was much larger than that of the control. Referring to the fatty acid composition of lipids in the literature, abnormalities of the fatty acid concentration, namely a high percentage of short-chain fatty acids of sulfatide and ganglioside in the cerebral cortex of the present cases, were confirmed. PMID- 2561571 TI - Pathological observations in HTLV-I associated myelopathy. AB - The following are the clinical and autopsy findings in a 63-year-old woman with myelopathy associated with the human T-cell lymphotropic virus Type I (HTLV-I). HTLV-I antibody was positive in both the serum and cerebrospinal fluid (CSF). In the lower thoracic region, demyelination and the loss of axons were accompanied by a proliferation of astrocytes, and gliosis was found in the lateral columns. Perivascular and parenchymal infiltrations of macrophages, lymphocytes, and plasma cells were also observed, but neither viral inclusion bodies nor atypical lymphocytes were found. PMID- 2561572 TI - Involvement of E. coli dcm methylase in Tn3 transposition. AB - The effects of DNA methyltransferases on Tn3 transposition were investigated. The E. coli dam (deoxyadenosine methylase) gene was found to have no effect on Tn3 transposition. In contrast, Tn3 was found to transpose more frequently in dcm+ (deoxycytosine methylase) cells than in dcm- mutants. When the EcoRII methylase gene was introduced into dcm- cells (E. coli strain GM208), the frequency of Tn3 transposition in GM208 was dramatically increased. The EcoRII methylase recognizes and methylates the same sequence as does the dcm methylase. These results suggest that deoxycytosine methylase modified DNA may be a preferred target for Tn3 transposition. Experiments were also performed to determine whether the Tn3 transposase was involved in DNA modification. Plasmid DNA isolated from dcm- E. coli containing the Tn3 transposase gene was susceptible to ApyI digestion but resistant to EcoRI digestion, suggesting that Tn3 transposase modified the dcm recognition sequence. In addition, restriction enzymes TaqI, AvaII, BglI and HpaII did not digest this DNA completely, suggesting that the recognition sequences of TaqI, AvaII, BglI and HpaII were modified by Tn3 transposase to a certain degree. The type(s), the extent and mechanism(s) of this modification remain to be investigated. PMID- 2561573 TI - A human-human hybridoma which produces antibodies to polioviruses. AB - A human-human hybridoma which produced antibodies to Sabin strain poliovirus types 3 and 2 was established. This hybridoma was constructed by fusing a HAT sensitive variant of a human lymphoblastoid subline R4-4-6 with peripheral blood lymphocytes of a donor who had been previously immunized with poliovirus Sabin vaccines and had shown high titer of neutralizing antibodies against these viruses. This hybridoma had been cultured for more than one year and the neutralizing activities of the culture supernatants against poliovirus type 3 could still be detected. Cross neutralization with poliovirus type 2 was also observed. Karyotypic analyses of this hybrid showed that the chromosome numbers were distributed mainly between 80 and 100. Dot immunobinding assays also confirmed the reactivities of this hybridoma supernatants with poliovirus type 3 and, slightly, with type 2. PMID- 2561574 TI - Production of transgenic mice carrying the human hepatitis B virus or the human papillomavirus DNA sequences in Taiwan: analysis of physical structure and hereditary mode. AB - In order to produce suitable animal model systems for the analysis of possible synergistic interaction between hepatitis B virus (HBV) or the human papillomavirus (HPV) with other viral or chemical co-factors, we have produced transgenic mice carrying HBV, HPV16 or HPV18. Transfer of 3343 micro-injected one cell (C57/BL6 X CBA) F1 embryos into pseudo-pregnant ICR recipients produced 181 pups. Seventeen of 151 of the pups analysed were found to be transgenics. Southern blot and slot blot analysis showed that the copy number of the transgenome in the transgenic mice ranged from 1-50 copies per cell. All the transgenomes analysed appeared to be in tandem repeats without detectable sequence rearrangements. The transgenome sequences were also stable on transmission to the F3 generation. On breeding, six of the ten transgenic mice analysed showed the Mendelian mode of inheritance with integration at single or multiple loci. Of the remaining four mice, three were mosaics and one was sterile. Analysis of the gene expression of the transgenic mice is currently in progress. PMID- 2561575 TI - Surgical treatment of angiofibromas of the nasopharynx--34 cases. AB - Between 1966 and 1987 we have treated 34 angiofibromas. The patients were between 9 to 28 years of age (average 16.5 years). The tumour extension was determined formerly by tomography and angiography, but these two methods have been replaced by angioscanning, which give the best results. Nineteen patients were classified stage I, fourteen stage II and one stage III. The surgical approach was transmaxillary in most cases. After tumour resection, two local recurrences were seen and a second operation was necessary to achieve a definitive cure in these patients. PMID- 2561576 TI - Juvenile angiofibroma--imaging techniques in diagnosis. PMID- 2561577 TI - [Determination of the dietary fiber content of some foods using a biological method]. AB - The evaluation of a biological method for dietary fibre determination in food was the main purpose of this study. Three-week old Wistar rats were used in this experiments. The rats were administered during two weeks a diet containing 1% of crude fibre from bread, grits, green beans, white cabbage and carrot. It was found that the biological method gave results similar to those obtained by the enzymatic one. PMID- 2561578 TI - Kinetics of 3H-thymidine label in rat liver regenerating after partial hepatectomy and after damage with carbon tetrachloride or silica. AB - Rat liver DNA was labelled with [methyl-3H] thymidine after partial hepatectomy, carbon tetrachloride poisoning, or an intravenous injection of silica dust. Changes in DNA labelling were studied for 4 weeks after the single pulse. Total radioactivity incorporated into liver DNA after partial hepatectomy and after carbon tetrachloride administration remained on the same level when compared with that found after 1 h. DNA activity in liver of untreated rats and of rats treated with silica decreased by about 50% within the first 2 weeks and then remained on this level for the rest of the studied period. These differences may reflect the fact that hepatocytes that have a long life span are preferentially labelled in partially hepatectomized and CCl4-treated rats, while liver macrophages with a short half-life take up a large part of the label in intact rats and in rats treated with silica. PMID- 2561579 TI - Seroepidemiological study of Epstein-Barr virus infection in children in Taipei. AB - SerumIgG antibody titer against viral capsid antigen(VCA) of Epstein-Barr virus (EBV) was measured by indirect immunofluorescence antibody method in 1350 children under 15 years of age, 723 males and 627 females, who were living in the Taipei area. Sera were collected from June through October 1984. Under the age of 6 months, the prevalence rate of positive serum VCA antibody was 38.4%. It decreased to 6.4% between 6 and 12 months of age, but rose to 65.8% during the second year of life. The rate continued to increase from 1 to 4 years of age, with a steep rise of geometric mean titer in the same period. The prevalence rate stayed at more than 90% after 4 years of age. From our data, it is obvious that most children in the Taipei area had been infected by EBV in early childhood, especially from 1 to 4 years of age. PMID- 2561580 TI - Perinatal cytomegalovirus infection in very low birth weight infants. AB - With the improvement in the survival of very low birth weight (VLBW, birth weight less than 1500gm) infants, problems specifically related to this weight group infants have been revealed. To define the incidence, morbidity and the possible role of blood transfusion of perinatally acquired cytomegalovirus (CMV) infection, 89 VLBW infants who had no evidence of congenital CMV infection were enrolled in a prospective study. 94% of the 82 infants tested had transplacental CMV IgG antibody at birth. According to the need of blood transfusion, Infants were divided into two groups: transfusion group (TF group) and nontransfusion group (NT group). Fifty two cases finished the study, thirty cases in the TF group, twenty-two cases in the NT group, and were subjected to the final analysis. Infants in this two groups were compatible in birth weight, gestational age, sex and mode of delivery. Infants in the TF group were sicker and had higher incidence of morbidity. Infants in the TF group received fresh blood from walking donors for small amount of blood transfusion. Each infant received an average of 52.3 +/- 42.6 ml blood from 2.3 +/- 1.3 donors. Ninety-five percent of the blood donors were CMV seropositive. Twenty out of the 52 (38.5%) VLBW infants developed perinatal CMV infection. Higher rate of perinatal CMV infection was found in the TF group (50.0%), comparing with the NT group (22.7%), p less than 0.05. More infants with low maternal antibodies at birth got perinatal CMV infection than infants with high maternal antibodies (47.6 vs 20%). Yet, the difference didn't reach statistical significance (p greater than 0.05, less than 0.1).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561581 TI - Neuromyelitis optica (Devic's disease) report of one case. AB - Neuromyelitis optica also named Devic's disease is an acute combined optic neuritis and transverse myelitis. It is thought to be a variant of multiple sclerosis, but its clinical presentation probably has only one attack without further recurrence and exacerbation. We present a 12-year-old girl who suffered from sudden onset of lower extremeties weakness, sensory loss and blurred vision after a prodromal URI symptom. CSF examination showed mild pleocytosis, elevated immunoglobulins, mild elevation of protein concentration. No oligoclonal band was detected. Serum virology showed high titer of anti-EB virus antibody. Visual evoke potential showed prolong of latency and decreased amplitude of both eyes. After prednisolone treatment, her visual accuity began to improve on the 7th hospital day and motor function improved on the 11th hospital day. Two years later, she has normal visual accuity, normal motor function and shows no evidence of disease recurrence. PMID- 2561582 TI - [Opportunistic infections of the central nervous system in patients with AIDS]. AB - Between August 1985 and March 1989 we had evaluated 70 AIDS patients. We had identified 7 opportunistic Central Nervous System infections, others toxoplasmosis and tuberculosis: A case of progressive multifocal leukoencephalopathy presented with aphasia as their initial manifestation, a probable herpesvirus type 1 polyradiculoneuropathy, four cryptococcal meningitis and syphilitic neuroretinitis. PMID- 2561583 TI - [Non-infective neurologic complications associated to heroin use]. AB - The spectrum of neurological complications associated with heroin addiction has changed in the past six years because of the progressive knowledge of the neurological complications related to HIV infection. We reviewed 48 heroin addicts with neurological complications and 452 heroin overdose who were seen in the Emergency Unit of our hospital during 1988 and the publications since 1967. Regarding the overdose we present the results of a prospective study leading to determine the causes. We emphasize the relationship with the level of total morphine in serum, instead of conjugate morphine, and with the presence of high levels of benzodiazepines found in the plasma rather than an hypothetic hypersensitivity phenomenon. We resume the neurological complications related with heroin addiction: spongiform leukoencephalopathy, epileptic seizures, stroke, transverse myelopathy and neuromuscular complications such mononeuropathy, plexopathy, acute inflammatory demyelinating polyradiculoneuropathy, rhabdomyolysis, fibrosing myopathy, musculoskeletal syndrome and acute bacterial myopathy. Some of such complications (i.e. transverse myelitis, polyradiculoneuropathy, leucoencephalopathy) must rise the suspicion of an HIV infection. Likewise, in patients assisted for overdosage we believe it's necessary rule out myoglobinuria by means of CPK serum levels and detection of urine hematic pigments without red blood cels in the urine sediment, in order to prevent and treat the renal failure. We report the results of muscular biopsy found in the musculoskeletal syndrome, which are similar to those found in alcoholic myopathy. Finally, we describe the clinical and diagnostic aspects in an unusually neuromuscular complication: the acute bacterial myopathy. PMID- 2561584 TI - [Neuropathology of HIV infection]. AB - Some references about the important neuropathologic lesions observed in patients with AIDS and about the variation on the relative incidence of the processes detected when analyzing clinic and neuropathologic tables, as well as a mention to the confusion created by terminological problems, clinicopathologic disagreement and wrong etiologic attributions, are used as an introduction to the exposition of HIV infection neuropathology. In a first section, Central Neuropathology is described, starting with the disorders considered as primarily caused by HIV. Then, infectious secondary Neuropathology, caused by different viruses, parasites, fungi or bacteria, and neoplastic secondary Neuropathology, mainly represented by localized lymphoma and by the uncommon Kaposi's disease, are analyzed. This section also includes references to cerebrovascular complications, Pediatric Neuropathology and ends with an analysis of cerebral biopsy indications. In the second part, Peripheral Neuropathology is described, splitting up peripheral nerve and muscle pathology. PMID- 2561585 TI - [Peripheral neuropathy in HIV infection]. AB - Neuropathy may complicate all stages of human immunodeficiency virus infection (HIV). Different types of peripheral neuropathy and myelopathy have been reported associated with HIV infection: sensory symmetrical polyneuropathy, acute inflammatory demyelinating polyneuropathy, chronic inflammatory demyelinating polyneuropathy, mononeuropathy multiplex, sensory ataxic neuropathy (ganglioneuronitis), cauda equina syndrome, amyotrophic lateral sclerosis, spastic paraparesia, and subclinical neuropathy diagnosed by electrophysiologic study. We describe the main clinical, electrophysiological and pathological features in these different types of neuropathy and comment their pathogenesis and treatment. Results in our series of twenty-two patients are also reported. In this series we want to underline three cases in which a chronic demyelinating polyneuropathy was the first manifestation of HIV infection. Thus, patients with predominantly motor demyelinating neuropathies and suspicious risk factors should be screened for silent HIV infection. PMID- 2561586 TI - [Neurophysiologic study in patients with human immunodeficiency virus infection]. AB - Peripheral nervous system (PNS) involvement in the form of different neuropathies in human immunodeficiency virus (HIV) infection is increasingly more common. We present a series of 20 HIV + patients subjected to neurophysiological study. The most commonly observed neurological disorder was a slight axonal-type sensitivo distal polyneuropathy that predominantly affected the lower limbs during the early stages of infection. As HIV infection evolves, the polyneuropathy worsens and becomes a sensitivo-motor polyneuropathy with a more important axonal component. Secondary demyelinization is observed, with additional involvement of the upper limbs in the final stages of infection. The most frequent neurophysiological finding was a decrease in somatosensory evoked potential (SEP) and conduction velocity (CV) of the sural nerve. PMID- 2561587 TI - Decentralization of maternity care. AB - One of the main reasons for the high maternal mortality rates in developing countries is the need to transfer patients long distances to city hospitals when they develop complications. Local obstetric facilities would relieve this situation. PMID- 2561588 TI - Photoaffinity labeling of the guinea pig pulmonary mast cell beta-adrenergic receptor. AB - Beta-adrenergic agonists can prevent mediator release from guinea pig pulmonary mast cells. By pharmacologic characterization, this response is mediated through a beta-2 receptor. Structural characterization of this receptor on the lung mast cell, however, has been limited by methods for isolation of this pulmonary cell. In this study, the guinea pig lung mast cell was isolated to greater than 90% purity, and its beta-adrenergic receptor identified by photoaffinity labeling with [125I]iodoazidobenzylpindolol (125IABP) and separation of membrane proteins by SDS-PAGE. We found the guinea pig pulmonary mast cell beta-adrenergic receptor to electrophorese as a heterogeneous protein between 68 and 116 kD. Photoaffinity labeling with 125IABP was protectable by alprenolol and isoproterenol but not by phentolamine and norepinephrine. Using subtype-selective compounds, the pulmonary mast cell receptor was established to be of a beta-2 subtype. This is the first report of the structural identification of a lung mast cell beta-adrenergic receptor and the first report of a beta-adrenergic receptor of approximately 100 kD in mass. This mast cell receptor is considerably larger than the 65 kD beta adrenergic receptors that have been identified in whole lung and other tissues. Data we have obtained using Northern blot analysis of mast cell RNA suggest a protein message of 45 kD for this beta-adrenergic receptor and a high degree of glycosylation most likely accounts for the large molecular size observed. PMID- 2561589 TI - Human alveolar macrophages release a factor that inhibits phagocyte function. AB - Human alveolar macrophages release in vitro a factor that inhibits both random migration and chemotaxis of human polymorphonuclear neutrophils (PMN). This factor is not cytotoxic and is recovered in culture supernatants of alveolar cells from most nonsmoking normal subjects. The inhibitor can be detected 30 min after cell cultures are established and is still produced after 24 h in culture. Its release was inhibited by cycloheximide. When supernatants are separated by molecular sieving (I-60 Waters HPLC column), most of the inhibitory activity is recovered in the low-molecular-weight fractions of the chromatogram (less than 1,000 D). The inhibitor has a broad spectrum of activity against known chemoattractants in that it reduces significantly the chemotaxis of PMN induced by the formyl peptide FMLP, by the complement fragment C5a, and by leukotriene B4; it also decreases the chemotactic activity associated with a monocyte-derived interleukin 1 preparation and the chemotactic activity derived from alveolar macrophage culture supernatants. The inhibitory factor is partially heat labile, is sensitive to aminopeptidase M, and is nonpolar. Both phorbol myristate acetate (PMA) and FMLP-induced superoxide release by PMN are diminished significantly in the presence of this inhibitory factor (p less than 0.01 for PMA and p less than 0.05 for FMLP). The inhibitor also reduces monocyte chemotaxis but has no effect on monocyte random migration. Finally, studies with [3H]FMLP indicate that this inhibitor does not act at the site of receptor binding on PMN. Thus, human alveolar macrophages can release in vitro both neutrophil chemotactic factors and an apparent neutrophil-inhibiting factor that may modulate positively and negatively the movement and the respiratory burst of neutrophils in the alveolar space. PMID- 2561590 TI - Neutrophils show chemotaxis to type IV collagen and its 7S domain and contain a 67 kD type IV collagen binding protein with lectin properties. AB - Neutrophils were found to demonstrate chemotactic responses to pepsinized human placental type IV collagen and its purified aminoterminal 7S domain. The maximal chemotactic responses occur at approximately 400 ng/ml and approximately 30 ng/ml of type IV collagen and 7S collagen, respectively, and are similar in magnitude to the chemotactic response of neutrophils to 10(-8) M FMLP. Human leukemic cells of the HL 60 line display chemotaxis to type IV collagen and 7S collagen only after they are differentiated along the neutrophilic pathway with dimethyl sulfoxide. When detergent extracts of neutrophils are applied to type IV collagen Affi-Gel resin, a 67 kD protein is retained by the resin and is eluted with guanidine/octyl-beta-glucoside or lactose. This 67 kD polypeptide has an amino acid composition resembling the 67 kD component of the elastin receptor complex, displays immunologic cross-reactivity with antibody to the 67 kD component of the elastin receptor, and binds to elastin and laminin affinity resins. Neutrophil chemotaxis to type IV collagen and 7S collagen is selectively abolished by exposing the test neutrophils to lactose or elastin peptides. We conclude that neutrophils may migrate in vivo to proteolytic fragments of type IV collagen and that this response may be mediated by a lectin-like protein that is similar to the 67 kD component of the elastin receptor. PMID- 2561591 TI - Phorbol ester-induced inhibition of the beta-adrenergic system in pulmonary endothelium: role of a pertussis toxin-sensitive protein. AB - To investigate possible cellular mechanisms for how activation of protein kinase C inhibits the relaxation caused by isoproterenol, we studied the effect of the protein kinase C activator 4 beta-phorbol-12 beta-myristate-13 alpha-acetate (PMA) on the increase in cyclic AMP (cAMP) production and adenylate cyclase activity caused by isoproterenol in bovine pulmonary artery endothelial cells. Treatment of intact cells with PMA prevented in a time- and dose-dependent manner the increase in cAMP production caused by isoproterenol, whereas 4 alpha-phorbol 12 beta-myristate-13 alpha-acetate (4 alpha-PMA), which does not activate protein kinase C, did not affect isoproterenol-induced cAMP production. PMA also reduced the increase in adenylate cyclase activity caused by isoproterenol, forskolin, and Gpp(NH)p. To test the hypothesis that the inhibitory effect of PMA is mediated via a pertussis toxin-sensitive G protein, we determined whether pretreatment of the cells with pertussis toxin would prevent the inhibitory effects of PMA. In pulmonary endothelial cells, pertussis toxin ADP-ribosylated an Mr 40,000 peptide that comigrated with the pertussis toxin substrate of human erythrocytes. Pertussis toxin treatment eliminated the inhibitory effect of PMA on isoproterenol-stimulated cAMP production and adenylate cyclase activity. Thus, the protein kinase C activator PMA inhibits the increase in cAMP production and adenylate cyclase caused by isoproterenol. This inhibitory effect in endothelial cells appears to be mediated via a pertussis toxin-sensitive protein. PMID- 2561592 TI - Cisplatin neurotoxicity. PMID- 2561593 TI - Radiation therapy for medically inoperable stage I and II non-small cell lung cancer. AB - The published results of primary radiation therapy for early stage NSCLC, indicate that it is a reasonable alternative in patients with medical contraindications or who refuse surgery, resulting in acceptable morbidity, local control, and survival rates. There is no conclusive evidence that EMI is of benefit. Consequently treatment with involved field alone, may be considered when there is no evidence of hilar involvement, or when it is necessary to limit the volume of lung tissue irradiated. Although the data are not conclusive, there is evidence to suggest that the total dose of radiation delivered to the primary should be sufficient to eradicate gross disease (60 Gy or higher). Such does result in high response rates particularly for T1 tumors. There is also an indication that complete responders have better survival than other patients, suggesting that radiotherapeutic strategies to enhance tumor eradication may improve survival. PMID- 2561595 TI - [Technology and progress in periodontics]. AB - This paper presents a discussion of some selected new technological aspects in periodontology; an evaluation of the progress in diagnosis and treatment of periodontal diseases today is attempted. The following aspects are considered: new developments in diagnostic radiography and alternatives, microbiological and immunological tests, clinical parameters, periotest method, tooth cleaning instruments, implantation of hydroxylapatite into periodontal defects, guided tissue regeneration, growth factors in periodontal treatment. PMID- 2561594 TI - Angiotensin converting enzyme inhibition, parasympathetic activity and exercise in essential hypertension. AB - Reduced parasympathetic activity has been reported in essential hypertension. Converting enzyme inhibition seems to increase parasympathetic tone. In order to evaluate the effects of enalapril on parasympathetic control of heart rate, the authors studied ten mild-to-moderate essential hypertensive patients (7 F, 3 M), treated for 2 weeks with placebo and for 1 month with enalapril. Compared to placebo, enalapril significantly reduced blood pressure (p less than 0.005 at least, both systolic and diastolic), without any change in heart rate. Enalapril enhanced parasympathetic activity as judged by the increased variation of heart period (VHP) during regular breathing. VHP was derived during continuous ECG recording by the difference between the mean of all maximum and minimum R-R intervals, taken as a measure of respiratory sinus arrhythmia: the higher the VHP, the higher the parasympathetic cardiac influence and vice versa. The response to exercise, used as an index of sympathetic stimulation, was not modified by enalapril: the heart rate peak reached during either static (hand grip) or dynamic (bicycle ergometer) exercise and the slope of the increase in blood pressure were unchanged. Therefore, enalapril appears to increase parasympathetic tone in essential hypertension, without any interference with sympathetic adaptation to stress. PMID- 2561596 TI - [Bone apposition following intraboney periodontal implantation of compact and phycogenic hydroxylapatite ceramic graft material]. AB - Intrabony periodontal lesions in three minipigs received surgical debridement followed by site implantation of synthetic and phycogenic hydroxylapatite graft material. Histologic examination of the sites treated with Allotropat 50 showed limited ossification of these implants. Lesions treated with the phycogenic material showed increased ossification of the implant pores and the implant periphery. These results suggest that Allotropat 50 has no clinical indication, while the phycogenic material could have a limited indication in intrabony periodontal lesions. PMID- 2561597 TI - The effect of a dentifrice containing soluble pyrophosphate and a copolymer on calculus deposit: a six-month clinical study. AB - A six-month, double blind, cross over clinical study was conducted to compare the effect of a commercially available anticalculus dentifrice and a placebo dentifrice on supragingival calculus deposits. Fifty adult subjects who entered the study were given an oral prophylaxis and were assigned to use either pyrophosphate/copolymer toothpaste or a placebo toothpaste for a period of three months. They were then scored for calculus deposits using the Volpe-Manhold method, received a second oral prophylaxis and used the alternate dentifrice for a second three month period of time. The subjects were again scored for calculus deposits and the study was completed. The results indicated that pyrophosphate/copolymer toothpaste reduced supragingival calculus formation by 37.07%, as compared to the placebo dentifrice. This reduction was significant at the 99% level of confidence. PMID- 2561598 TI - [Clinical application of sintered bone (II). Effects of collagen coating on implant materials in vitro]. AB - In vitro studies with osteoblast-like cells, revealed that in addition to preventing the immediate outflow and promoting stabilization of implant materials, these cells have the additional effect of promoting the mending and early calcification by coating the materials with collagen. The materials used for the experiment were minute hydroxyapatite, beta-tricalcium phosphate and bovine sintered bone. To these, atelocollagen which is solble in pepsin extracted from calf corium and crosslinked by using ultraviolet rays was added. We observed the cells very closely after these materials were added to alpha-modified eagle's medium containing 10 mM beta-glycero phosphate, 10% fetal bovine serum and osteoblast-like cells and the cultures incubated at 37 degrees C and 5% CO2. After 14 and 21 days, cells were fixed and stained with Alizarin Red S and Von Kossa stains to measure calcification. As a result of the collagen coating, positive areas appeared and, compared to the control improvement of the early mending was apparent. The results suggest that by coating the materials with collagen, the osteoblast-like cells show better mending and early calcification. PMID- 2561599 TI - [Application of calcium phosphate ceramics to periodontal therapy. 8. Effects of orthodontic force on repaired bone with hydroxyapatite]. AB - It has been unclear whether M.T.M. is acceptable for patients treated with bone grafts using hydroxyapatite (HAP), which is not biodegradable. Therefore we studied histological changes in bone among HAP during orthodontic tooth movement. HAP was implanted into artificial bone defects adjacent to second premolars in beagle dogs, and as a control no material was implanted Three months after implantation, the second premolars were moved with orthodontic force. Then they were observed histopathologically. The results obtained were as follows. The amount of movement of the teeth in implanted sites was less than in the controls. In the pressure zone, resorption of bone by osteoclasts was observed in both the implanted and the control sites, in addition, root resorption occurred adjacent to HAP. Furthermore multinucleated giant cells were closely attached to HAP and were surrounded by bone and connective tissue. In retention period there was ankylosis between the roots and bone around HAP. In the tension zone, additional bone from HAP was observed. Bone defects were filled with bone in the implanted sites, but with connective tissue in the controls. These results suggest that bone among HAP was resolved by osteoclasts. Clinically, HAP implantation for the pressure zone should be avoided because of the occurrence of root resorption and ankylosis. But for the tension zone it seems to be effective because HAP implantation produces more repaired bone. PMID- 2561600 TI - [Application of synthetic hydroxyapatite to periodontal therapy]. AB - The purpose of the present study was to evaluate the effectiveness of granular hydroxyapatite as an alloplastic bone implant material in infrabony periodontal defects. Thirty-five infrabony defects in 28 subjects with marginal periodontitis were used. The patients were instructed in oral hygiene and received several sessions of scaling and root planing. Granular hydroxyapatite was implanted into test sites with infrabony defects following application of internally beveled full thickness flaps, root planing, and defect debridement. Pocket depth and attachment level were recorded before and 3 and 6 months after surgery. Gingival recession, mobility of the tooth, plaque index, and gingival index were also recorded before and 1 and 2 weeks, and 1, 3 and 6 months after surgery. Redness and swelling of the gingiva and increase in tooth mobility appeared transiently after surgery, however, they disappeared in a short time. A significant reduction of mean pocket depth of 3.4 mm and a significant increase in the mean attachment level of 2.5 mm were observed at 6 months after surgery. The mean gingival recession during the experimental period was only 0.9 mm. The amount of hydroxyapatite remaining implanted in the infrabony defect was 73% 6 months after surgery. The data and clinical impression suggest that hydroxyapatite has a potential as an alloplastic implant with clinically apparent acceptance by soft and hard tissue. PMID- 2561601 TI - [Hydroxyapatite implant for human periodontal osseous defects]. AB - The purpose of this study was to evaluate the efficacy of porous hydroxyapatite (HAP) as an implant material into human periodontal defects. Thirty three patients affected by marginal periodontitis were studied. Following initial therapy, 44 periodontal osseous defects received grafts of HAP granules and blocks during flap surgery. Radiographic and various clinical examinations were performed before surgery and 1 and 2 weeks, and 1, 2, 3, 6 and more than 6 months after surgery. The results were as follows: 1. Postsurgical inflammation of the gingiva disappeared in a short time. Further more open wounds and discharges of HAP disappeared in the first month. There were no clinical problems in the sites receiving HAP implantation. 2. Radiographic demarcation lines between the implanted HAP and the surrounding alveolar bone became unclear at the beginning of bone repair. 3. At six months after surgery, the mean values of gingival recession, decrease in pocket depth and clinical attachment gain were respectively 0.9 mm, 2.7 mm, 1.9 mm. Additional observation beyond 6 months showed an attachment gain of 2.0 mm. 4. The attachment gain was less in the cases of one wall osseous defect and of furcation involvement in lower molars. These results suggest that HAP implantation is clinically effective, however, careful application is needed for the cases of one wall osseous defect or furcation involvement. PMID- 2561602 TI - [Evaluation of hydroxyapatite implants in human periodontal osseous defects]. AB - In this clinical study we evaluated the use of hydroxyapatite ceramic (HAP) implant in conjunction with the surgical treatment of periodontal osseous defects. Fifty-five defects in 23 patients were treated and filled with HAP particles (BDHAP-101). At 6 months and 12 months after the placement of particles, the sites were evaluated by gingival margin heights, periodontal pocket depths, attachment levels, tooth mobility, the gingival index (GI), and radiographic analyses. Oral hygiene status was estimated by the plaque index (PlI). There were significant improvements with regard to probe depth, attachment level, tooth mobility, and GI. The radiographic analysis showed a similar density of the adjacent bone and the particles, suggesting excellent biocompatibility. There were no changes in PlI before or after the placement of implants. PMID- 2561603 TI - [Histological study of multinucleated giant cells in bone grafting]. AB - The purpose of this study was to determine histochemically the characteristics of elicited multinucleated giant cells adhering artificial graft materials. As artificial graft materials, resorbable ceramic beta-tricalcium phosphate, non soluble porous hydroxyapatite and non-soluble non-porous hydroxyapatite were examined, with devitalized bone particles serving as the control. Both test and control graft materials were implanted subcutaneously in rats and mice. Acid phosphatase and tartrate resistant acid phosphatase were positive in elicited multinucleated giant cells adhering to both artificial graft materials and devitalized bone particles. Elicited multinucleated giant cells elicited adhering to sutures were negative. The number of multinucleated giant cells positive to non-specific esterase was minimum except in areas they adhered to sutures. Elicited multinucleated giant cells adhering to graft materials showed enzyme activities similar to those of osteoclasts, and it was suggested that artificial graft materials may be absorbed by osteoclasts. PMID- 2561604 TI - [Effect of hydroxylapatite particles during healing of experimental furcation involvement in beagle dogs]. AB - The present study was performed to evaluate the effects of hydroxylapatite (HAP) on tissue regeneration in various types of furcation involvement. Upper premolar tooth sites of 9 beagle dogs (3-6 years old) were used. HAP particles were implanted into three types of furcation defects: Class III lesion (by Glickman) in 2nd premolar sites, artificially caused by intrafurcal suturing or spontaneously developed about 3 months after extraction of the 1st premolar; Class IV lesion in 4th premolar sites artificially produced 2 months before implantation; and through-and-through furcal bony defects in 3rd premolar sites at the time of HAP implantation. Macroscopic, radiographic and microscopical investigations of the postoperative status were carried out at intervals of 1, 2, 3 and 6 months. The appearance of the gingiva in bony defect sites was almost the same as that in the presurgical status 2 weeks after implantation, although gingival inflammation was persistent in Class III and IV lesions. Most particles in Class III and IV lesions exfoliated until 2 weeks after implantation, and the junctional space between recipient bone and particles could not be distinguished, probably as a result of incorporation of HAP and osseous tissue. Histological observation revealed that HAP particles were surrounded by new bone located at the top of the alveolar crest in all defects. However, there were no obvious signs of coronal bone formation or connective tissue attachment in Class III and IV lesions up to 3 months after the operation. At 6 months there was evidence of new bone formation over the presurgical crests in Class III and IV lesions. On the other hand, there were obvious signs of connective tissue attachment and bone formation in bony defect sites, probably induced by the tissue of the periodontal ligament remaining after the surgical procedure, and nonphysiological ankylosis between root and bone was frequently observed. There were notable findings, such as newly formed osteoid tissue intervening with the adhesive particles, calcified tissue intervening between HAP particles and root surface showing ankylosis, and peripheral osteoid formation in the HAP particles. It is postulated that HAP has no apparent role in induction of bone formation, although there is chemical affinity to calcified tissue, and it is effective in yielding a volume of bone like tissue where osseous repair could be performed. However, HAP did not enhance regeneration of lost periodontal structures including connective tissue attachment. PMID- 2561605 TI - Effect of lead acetate administration on ATPases of brain and its mitochondrial and synaptosomal fractions in adult mice. AB - The effects of oral administration of lead acetate on the activities of cation transport ATPases and on the brain and its mitochondrial and synaptosomal fractions of male mice were studied at doses of 1, 5, and 20 mg lead acetate per 100 g body weight per day for 4, 8, 12, and 16 weeks. The activities of Na(+) ATPase, K(+)-ATPase, total-ATPase, and Na(+)-K(+)-ATPase decreased significantly at doses of 1 and 5 mg lead acetate after 12 and 16 weeks of treatment; changes in the activities were not marked after 4 and 8 weeks of lead administration. However, the 20-mg dose significantly reduced enzyme activities at all treatment intervals. PMID- 2561606 TI - [Many-year survival after surgical treatment of glioblastoma multiforme. Report of 2 cases]. AB - Two female patients are described with survival over 13 years after operation for glioblastoma multiforme. The first patient was 42-year-old at the time of partial removal of the tumour situated in deep parts of the temporal lobe. After the operation she was not given any radiotherapy. CT done 13 years after the operation failed to show tumour presence. The patient is leading a self-dependent life (80 points in Karnofski scale). The other female patient was 28-year-old at the time of nearly complete removal (macroscopic) of right temporal lobe tumour. She received cobalt radiotherapy. CT 9 years after the operation showed no tumour. The present state of the patient was evaluated at 90 points Karnofsky scale. In no case cytostatics were given. These cases demonstrate an exceptionally long survival after operation for malignant glioma. The cause of this long survival is not known. PMID- 2561607 TI - [Ductal carcinoma of the breast. Electron microscopic study]. AB - Electron microscopic study of the breast tumors were carried out on 67 patients. There were 7 intraductal carcinomas and 60 infiltrating ductal carcinomas of the breast. Observations were made on the morphological characteristics of neoplastic cells, myoepithelial cells and basal lamina. Cells organelles in the neoplastic cells are similar in both types of ductal carcinomas. Nuclei are predominantly large and irregular with poor dense chromatin. Often a lot of filaments are around the nuclei. Myoepithelial cells in the infiltrating ductal carcinomas are ultrastructurally the same as seen in the intraductal carcinomas, but they seems to be less numerously than in the intraductal carcinomas. In the intraductal carcinomas of the breast basal lamina is irregular and from time to time is disrupt. In the infiltrating ductal carcinomas basal lamina is usually absent, but sometimes we can find its short fragments. PMID- 2561608 TI - [Trophoblastic tumor at the site of the placenta]. AB - The author describes the histological structure of a rare placental site trophoblastic tumor. In differential diagnosis it is necessary to take into account hydatidiform mole and chorionepithelioma. In differentiating with hydatidiform mole lack of placental villi is characteristic while with choriocarcinoma the presence of connective stroma in PSTT is helpful. PMID- 2561609 TI - [Mixed mesodermal tumor of the uterus]. PMID- 2561610 TI - Observations on the immunology of small cell lung cancer with implications for future management. PMID- 2561611 TI - [Histological types of lung cancer in urban and rural inhabitants of the Krakow region]. AB - Incidence of different histological types of lung cancer was analyzed in the years 1978-1984 in city and rural inhibitants of the Cracov region. Histologically verified lung cancer ranged from 21% in females of rural population to 52% of city male inhabitants. Squamous lung cancer followed by small cell lung cancer were most often diagnosed in city population and males from villages. Adenocarcinoma was seen more often in females, being most often diagnosed in females of rural population. PMID- 2561612 TI - [Bronchial carcinoid and small cell lung cancer--neuroendocrine tumors. Immunohistochemical studies]. AB - Selected neoplastic markers (NSE, gastrin, CEA, calcitonin, keratin) were studied in pulmonary specimens from 5 patients with bronchial carcinoid, 20--with small cell lung cancer (SCLC), and 2 with solid tumors. In patients with carcinoid and SCLC NSE and gastrin markers were found--characteristic for neuroendocrine neoplasia. The author discuss the usefulness of immunohistochemistry in differential diagnostics of pulmonary malignancy. PMID- 2561614 TI - Micromorphologic features of dentin in vitamin D-resistant rickets: correlation with clinical grading of severity. AB - This study was a histological analysis of 20 primary teeth from 5 patients with 3 clinical grades of vitamin D-resistant rickets (VDRR). The results showed that the degree of globular dentin formation in the histological sections may be graded into Grades I-III in increasing order of severity. In Grade I, the amount of globular dentin was less than 50% of the total dentin thickness, and the interglobular spaces were small. By contrast, in Grade III severity, globular dentin extended throughout the entire thickness of dentin, and the interglobular spaces were large. In Grade II severity, the amount of globular dentin was more than half but did not involve the entire dentin thickness. These histologic grades of severity correlated directly with the clinical grades of the patients. In addition, the study found that lack of medical treatment in affected mothers might lead to globular dentin formation in the fetus in-utero. Conversely, adequate phosphate supplementation in a hypophosphatemic mother might prevent the formation of globular dentin in the fetus. PMID- 2561613 TI - [Long-term survival of patients with small cell lung cancer]. AB - The analysis of clinical determinants of long-term survival in small cell lung cancer was investigated in consecutive series of 469 patients included in prospective multicenter clinical trials from 1981 to 1985. Forty eight patients (19.2%) were alive after 2 years from initiation of therapy and among them 27 (5.8%) were disease free. The most important clinical determinants of long-term survival were: extent of disease, performance status and sex. 38 out of 243 patients with limited disease (15.6%) survived for 2 years or more as well as 10 out of 226 patients with extensive disease (4.4%, p less than 0.001), 33 out of 237 patients with WHO performance status 0 and 1 (13.9%), and 15 out of 232 patients with performance status from 2 to 4 (6.4%, p less than 0.01), 29 out of 229 (12.2%) with absence of weight loss before therapy and 19 out of 240 (7.9%) with weight loss (N.S.), 32 out of 392 males (8.2%) and 16 out of 77 females (20.7%, p less than 0.01). Out of 27 disease-free survivors 21 are alive with no sign of malignancy after 3.5 to 7 years from initiation of therapy. Ten patients out of 229 followed up for a minimum 5 years after inclusion to the studies survived this period with no signs of disease. This study confirms the possible curability of small cell lung cancer, especially in patients with favorable prognostic characteristic. PMID- 2561615 TI - Pattern of acquisition of rotavirus antibody in children followed up from birth to the age of three years. AB - Nine hundred and forty-eight serum samples from 83 children living in Belem, Brazil, collected within their first three years of life, were tested for the presence of group-specific rotavirus-antibody by an enzyme-linked immunosorbent assay (ELISA) blocking-test. Passively transferred maternal antibody lasted about two and half months; subsequently, low levels of rotavirus antibody started to appear at seven months, reaching a peak at eleven months of age. From one year onwards positivity gradually increased, reaching highest values at 34 months of life. Individual responses were examined in sera from 61 children who were followed up since birth to three years of age: 38 (62.3%) of them developed a long-term immunity following first infection; eleven (18.0%) children developed a short-term immunity after first infection by rotavirus; seven (11.5%) had no antibody response within their first three years of life; and 5 (8.2%) showed positive antibody response from birth to three years old. PMID- 2561616 TI - [Evaluation of conductivity in motor nerve fibers of the ulnar nerve in syringomyelia and neuropathy using 6 stimulation indicators]. AB - The ulnar nerve's motoric fibre conduction velocity was stated to be insignificantly disordered in 32 patients with syringomyelia, whereas 47 neuropathic patients with unknown etiology showed the disorder mentioned above as occurring by far more frequently and in a more extent. From 6 stimulative indices, those of distal extremities velocity had maximum of deviations. Nevertheless, the indices of longer sections revealed the most numerous pathologic findings, especially in determining forearm velocity. The obtained findings are compared with 233 control subjects. PMID- 2561617 TI - [Study of conductivity of the motor fibers of the ulnar nerve with respect to intensity, localization and diagnosis of peripheral nerve disease. I. General and morphologic observations on the affected area of the ulnar nerve with emphasis on the elbow region and causes of the disease]. AB - The introductory part deals with a great community impact of ulnar nerve damage and its high incidence. From peripheral nerves, this ulnar is the most frequently exposed to injuries and impresses. The impressory damage occurs mainly in elbow region. Anatomic peculiarities of ulnar nerve are scrutinized concerning with more connective tissue presence on its cutting section comparably to the other upper limb nerves. Frequent innervative deviations are referred to especially in ulnar and median nerve regions in the course of ulnar nerve branching. Also three types of anastomoses are listed between the both nerves which are important as to the clinical pattern in damages and for electrophysiologic diagnosing. From the clinical tests, those of rare use are emphasized (test of crossed fingers, palmaris brevis sign, Mumenthaler's sign, ulnar test). The last part deals in detail with anatomic, functional and clinical problems of ulnar nerve in elbow area. Its biomechanic relations in the cubital tunnel are elucidated as well as etiopathogenetic factors of neural damage in this area (traction and compressive theories and hypermobility of the nerve etc.). The most frequent causes of ulnar injuries are reviewed. The present work is of introductory value for those dealing mainly with electrophysiologic problems of both sensitive and motoric neural injuries varying in the intensity, location and diagnosis. PMID- 2561618 TI - [Microsurgical reconstruction of the mandible using a vascularized iliac bone graft]. AB - A defect in the mandible, which also included the mandibular process, was successfully repaired with a free vascularized iliac bone graft. The development of the procedure and the operative technique are discussed. Advantages and disadvantages are compared and reviewed on the basis of a pertinent case. Together with others reporting in the international literature the authors believe that this method is useful in the management of extensive defects, in which conventional bone grafting usually fails. PMID- 2561619 TI - [Sandwich osteotomy versus interpore implants for maxillary versus mandibular ridge augmentation]. AB - Horse-shoe sandwich osteotomy with interposed cancellous bone for augmentation of highly atrophic maxillas is described. 2-year results were encouraging in terms of denture fit, but long-term results are needed for a definitive assessment. For severely atrophic mandibles the use of porous hydroxyapatite (Interpore 200) is shown to be beneficial and prevents further bone resorption. After complete healing dislocation and loss of ridge height were absent, except in one special case. Subsequent vestibuloplasty for relative augmentation proved to be unnecessary. For a definitive assessment of this clinically successful method long-term results will have to be waited for. PMID- 2561620 TI - [Inhibition of hard dental plaques by reactive components of dentifrices]. AB - Active components of toothpastes which are intended to inhibit plaque mineralization are discussed with their physiological and chemical implications on the formation and calcification of dental plaque. The decisive factors are defined as part of the biocrystallographic reactivity in the oral milieu. PMID- 2561621 TI - [B-mode sonography in the diagnosis of unclear soft tissue masses of the face and neck]. AB - By definition, ultrasonography images the gross morphology of normal organs and of abnormal connective tissue structures. The site and demarcation of lesions, their topographical relations to other structures and the internal echo patterns are illustrated on the basis of seven clinical cases. The role of sonography in the diagnostic work-up is defined. The choice of treatment is determined by the early detection of a mass and its further appearance in terms of size and echogenicity. Sonography cannot replace histology. This is why so-called "typical" echo patterns may be misinterpreted. For follow-up studies sonography needs to be done in carefully defined planes. For ultrasonography to be helpful in the diagnosis, a full understanding of the patient's history and clinical signs and symptoms in the head and neck region is critical. Prior to sonography these data should, therefore, be elicited by the examiner himself. PMID- 2561622 TI - Chronic modification of dopaminergic transmission and its effects on binding characteristics of dopamine transporter ligand 3H-GBR 12783 in the striatum. PMID- 2561623 TI - The behavioural effects of benzodiazepine receptor ligands in mice are inhibited by adamantylamide dipeptide. PMID- 2561624 TI - A peripheral antagonist of benzodiazepine receptors (PK 11195) did not reverse behavioural effects of diazepam in mice. PMID- 2561625 TI - Heterogeneity of human monocytes: differences in response to IFN-gamma. AB - Human peripheral blood monocytes can be separated into two subpopulations which differ in the efficiency of their adherence to glass after 16 hours of incubation. The adherent subpopulation was found to be about twice as effective in binding mannose-resistant E. coli 0-124, mannose-sensitive E. coli 0-128 and opsonised E. coli than the nonadherent one. In addition, reduction of cytochrome C in response to E. coli binding or 12-myristate 13-acetate (PMA) stimulation was two fold higher in adherent cells. The binding of E. coli O-124 and the superoxide generation stimulated by E. coli were inhibited by the addition of mannose only in the adherent monocytes, indicating the presence of mannose receptors on the cell surface in the adherent subpopulation. The treatment of the nonadherent cells with 0.1-1000 U/ml of Interferon (IFN-gamma) for 24 hours resulted in a dose dependent increase in superoxide generation. After 72 hours of incubation with IFN-gamma (1000 U/ml) the amount of superoxide generated by the nonadherent cells was elevated to 20.5 +/- 1.4 nmoles/10(6) cells/15 min, similar to that of the adherent cells (24.5 +/- 1.2 nmoles/10(6) cells/15 min untreated adherent monocytes). The generation of superoxide in the IFN-gamma treated nonadherent monocytes stimulated by E. coli 0-128 was significantly reduced by addition of mannose. PMID- 2561626 TI - Corticotropin (ACTH) induction of tumor necrosis factor alpha by monocytes. AB - The finding that monocytes possess specific ACTH receptors led us to determine the effect of ACTH on monokine production. Here, we report that ACTH 1-39 will induce TNF-alpha from the adherent fraction of peripheral blood leukocytes. In addition, we also found that ACTH 1-39 will potentiate IFN-gamma's induction of TNF-alpha from these cells. Since previous reports showed that ACTH inhibits IFN gamma's activation of macrophages to a cytocidal state, our data raise questions concerning the relative role of TNF-alpha in macrophage mediated cytolysis. PMID- 2561628 TI - Surgical treatment of extensive, metastazing and small cell carcinoma of the lung. PMID- 2561627 TI - Induction of complement factor B activity in human fibroblasts by IL-6/IFN-beta 2 and IFN-gamma. AB - Human skin fibroblasts synthesize and secrete complement Factor B, a component of the complement alternative pathway, when stimulated by mediators of inflammation such as lipopolysaccharide and various cytokines. Recombinant IL-6/IFN-beta 2 (E. coli) stimulates Factor B synthesis in fibroblasts but the effect is strongly potentiated by the addition of IFN-gamma. When both cytokines are added, the skin fibroblasts secrete significant amounts of biologically active Factor B detectable in a hemolysis test. This cooperative effect of IL-6, which is made by most tissue cells and monocytes and of IFN-gamma which is made by T-lymphocytes may play a role in local inflammatory processes. IL-6 and IFN-gamma also cooperate in the induction of (2'-5') A synthetase, a mediator of IFN action. PMID- 2561629 TI - In vitro and in vivo studies on triclosan/PVM/MA copolymer/NaF combination as an anti-plaque agent. AB - A combination of triclosan, copolymer of PVM/MA and NaF has been optimized in vitro and in vivo for plaque effect. Triclosan (2, 4, 4' trichloro-2' hydroxydiphenyl ether), a non-cationic antibacterial, was compatible with anionic surfactant, NaF and PVM/MA copolymer as evaluated by in vitro antibacterial tests. The in vitro uptake of triclosan on the saliva-coated hydroxyapatite disks, extracted human teeth and oral buccal epithelial cells was enhanced two fold in the presence of PVM/MA copolymer when compared to the uptake in the absence of the copolymer. The combination consisting of 0.3% triclosan/2% PVM/MA copolymer/1100 ppm NaF was found to be highly effective on reducing smooth and fissure caries, and plaque extent compared to to 1100 ppm/NaF by itself in rats when applied topically. The combination also showed significantly more fluoride uptake and reduction in enamel solubility than 1100 ppm/NaF. The results of these studies indicated that this optimized combination was an effective antiplaque/anticaries agent in vitro and in vivo. PMID- 2561630 TI - Superoxide-independent hydrogen peroxide release by activated macrophages. AB - The present data show that freshly explanted BCG-activated mouse peritoneal macrophages release large quantities of hydrogen peroxide upon initial contact with a foreign substratum, without the requirement for other membrane stimuli such as phorbol diesters. The hydrogen peroxide detected under these conditions does not originate from extracellularly released superoxide, since 2 x 10(5) BCG activated macrophages spontaneously released 1.6 nmol hydrogen peroxide but only 0.2 nmol superoxide. Thus, more than 90% of the hydrogen peroxide detected was not derived from extracellular superoxide dismutation. The dissociation between hydrogen peroxide and superoxide release was further demonstrated in cytochalasin B- or lidocaine-treated cells or in the absence of glucose. Under these conditions, hydrogen peroxide release was markedly inhibited while superoxide release was unaffected. These observations provide evidence that another metabolic pathway is involved in the generation and release of hydrogen peroxide during adherence and spreading of freshly explanted activated macrophages onto a substratum. PMID- 2561631 TI - [Hepatocellular adenoma and focal nodular hyperplasia]. AB - The diagnostic findings of hepatocellular adenoma and focal nodular hyperplasia became more frequent in the last years in our as in western experience. The improvement in diagnostic technique, a correct pathological identification and the diffusion of oral contraceptives explain this increase of incidence. In our series were present 11 hepatocellular adenomas and 19 focal nodular hyperplasias: all the HCA cases were radically resected, while only 15 FNH were resected, only two biopsied and two submitted to periodical follow-up. The tendency to spontaneous bleeding and the presence of diagnostic uncertainty versus well differentiated hepatocellular carcinoma are mandatory indications for radical resections in all the HCA cases. FNH resection is reserved to symptomatic cases and wide wedge biopsy is at least required in presence of diagnostic doubts: the asymptomatic FNH ("hot spot" on Tc99m-HIDA scintigraphy) may be followed-up only. PMID- 2561632 TI - [Immunocytochemical demonstration of S-100 protein in adenoid cystic carcinoma of the salivary gland]. AB - Immunocytochemical study was performed for the presence of S-100 protein in adenoid cystic carcinoma (ACC) of salivary gland and normal salivary gland tissues with the immunogold-silver staining method (IGSS). Myoepithelial cells, intercalated duct cells and serous acinar cells of normal salivary gland were S 100 positive. In ACC, S-100 protein can be found in component cells of various histological patterns of the tumor. The results suggest, according to the distribution patterns of S-100 protein positive cells in the tumor tissues, that ACC may originate partly from intercalated duct cells and partly from myoepithelial cells. PMID- 2561633 TI - Antistress hormonal responses of analgesic nitrous oxide. AB - We briefly describe the hormonal responses associated with stress and provide evidence that endogenous and exogenous opioids have "antistress" hormonal profiles. We present data which indicate that analgesic nitrous oxide produces a typical opioid hormonal response, viz. increased prolactin and decreased cortisol levels. There was no significant change in ACTH levels despite the fall in cortisol. We present evidence which supports the hypothesis that the antistress hormonal effects of opioids and analgesic nitrous oxide are mediated by uncoupling the adrenal gland from hypothalamico-pituitary stimulation. It would appear that the opioid system acts antagonistically to the adrenocortical system in stress. We propose that any method stimulating the opioid system optimally may have the antistress effects. PMID- 2561634 TI - Mu and kappa opioid modulation of olfactory bulb evoked potentials. AB - Modulatory influences of the mu opioid agonist morphine and the kappa opioid agonist U-50, 488H on field-evoked potentials from the main olfactory bulb (MOB) in response to stimulation of the olfactory nerve were studied. Topically administered morphine upon the MOB dorsal surface produced a noloxone-sensitive depression of the late component of the response without modifying the early one, while topical administration of U-50, 488H suppressed both the early and the late components. Naloxone did not antagonize U-50, 488 effects. Results indicate that mu and kappa opioid agonists can interfere with sensory transmission at the level of second-order neurons of the olfactory pathway, the mitral and/or tufted cells. PMID- 2561635 TI - Immunogenicity in monkeys of two polio type 3 seed viruses (SO + 2 and SOR + 1) presently used for production of oral polio vaccine. AB - Rhesus monkeys were immunized orally and intramuscularly with polio type 3 seed viruses (SO + 2 and SOR + 1). No immune response was detected after oral feeding. Geometric mean titres of 891.4 and 237.02 were obtained after intramuscular administration of six and eight doses of SO + 2 and SOR + 1 respectively. All the monkeys immunized with SO + 2 and SOR + 1 were positive for antibodies after third and fifth dose respectively. High antibody titre (1:1024) was reached in 80 per cent monkeys after sixth dose with SO + 2 whereas this antibody titre was obtained in 20 per cent monkeys after eight doses of SOR + 1. SOR + 1 was found to be a poor immunogen as compared to SO + 2 as it had produced low titre antibody response in monkeys even after administration of eight doses. PMID- 2561637 TI - A heparin which binds to the envelope glycoprotein gp120 inhibits human immunodeficiency virus replication. PMID- 2561636 TI - [Presence of rotavirus and adenovirus in fecal samples of children with gastroenteritis, in the city of Goyania]. AB - In an attempt to detect rotavirus and adenovirus prevalence among other enteropathogens (bacteria and parasites) in diarrhoea, three hundred fecal samples originating from children living in Goiania city (Goias state, Brazil) were analysed. Rotavirus was found to be the only pathogen in 47 cases, and associated with other infectious agents in 21 cases. 97.0% positive samples of rotavirus showed an electrophoretic pattern characteristic of subgroup II. Adenovirus was found in 7 cases, and associated with other microorganisms in 1 case. Three methods were applied for virological analyses: enzyme immunoassay for rotavirus and adenovirus (EIARA), polyacrylamide gel electrophoresis (PAGE) and immunoelectron microscopy (IEM). The concordance among the three methods was 92.8%, PAGE and EIARA agreed in 95.8%, and IEM and EIARA agreed in 100.0%. PMID- 2561638 TI - Selective inhibition of thrombin- and plasmin-induced platelet aggregation by a synthetic peptide disulfide. AB - 1. A synthetic peptide disulfide, Gln-Val-Val-Cys(NpyS)-Gly-NH2 (P1) inhibited thrombin and plasmin-induced platelet aggregation and cleavage of aggregin. P1 did not inhibit platelet aggregation induced by other agonists nor did it inhibit shape change. 2. P1 also inhibited purified platelet calpain II. 3. The correspondence between the molecular structure of P1 and inhibitory sequence of the peptide in domain 2 of high molecular weight kininogen has shed light on the molecular nature of the cellular mechanism underlying thrombin- and plasmin induced platelet aggregation and the inhibition by P1. 4. P1 may prove to be useful in designing and improving future protocols of thrombolytic therapy to prevent reocclusion. P1 may also have a role in inhibiting thrombin formed during angioplasty and thus preventing restenosis. PMID- 2561639 TI - Net fluid secretion by mammalian renal epithelial cells: stimulation by cAMP in polarized cultures derived from established renal cells and from normal and polycystic kidneys. PMID- 2561640 TI - Purification and characterization of ceruloplasmin receptors. AB - Membrane receptors for ceruloplasmin were isolated by affinity chromatography using CP bound to CNBr-activated Sepharose 4B. The receptor was obtained in the form of a single sharp peak which when collected, concentrated and run on SDS PAGE provided a single band with a molecular weight of 35,000. Treatment with endoglycosidase F reduced this molecular weight by 3%. It is concluded that ceruloplasmin receptors in this cell system is a membrane protein containing 3% carbohydrate with a total molecular weight of 35,000. It is possible that this is a monomer of a higher molecular weight protein. PMID- 2561641 TI - Increases in levels of procollagenase mRNA in human fibroblasts induced by interleukin-1, tumor necrosis factor-alpha, or serum follow c-jun expression and are dependent on new protein synthesis. PMID- 2561642 TI - Herpes simplex virus is associated with peptic ulcer disease. PMID- 2561643 TI - The gene encoding alpha-galactosidase A and gene rearrangements causing Fabry disease. PMID- 2561644 TI - Protein I (Por) of Neisseria gonorrhoeae as an immunogen: liposomes, proteosomes, and the lack of blocking antibodies. PMID- 2561645 TI - Expression of human hypoxanthine guanine phosphoribosyltransferase mRNA in brains of mice infected with a recombinant herpes simplex virus type 1. PMID- 2561646 TI - [Anti-HBS, anti-CMV, and anti-HIV antibodies and HBS antigen in patients with hemophilia, malignant lymphomas, leukemias and multiple myeloma]. AB - In 21 patients with haemophilia, 10 with acute leukaemia, 12 with malignant lymphomas, and 12 with multiple myeloma in whom the risk of viral infection is increased the following antibodies were determined: anti- CMV, anti-HIV, anti HBs, and HBs antigen by ELISA test. Anti-CMV were found mainly in acute leukaemia (90%), in haemophilia (71.4%), in malignant lymphoma (41.7%) and multiple myeloma (33.3%). In 19% of cases of haemophilia anti-HIV antibodies were present. In other groups these antibodies were not found. In acute leukaemias mostly anti-HBs antibodies were present. The group of haemophiliacs is particularly exposed to infection by these viruses which is connected unquestionably to blood transfusions. PMID- 2561647 TI - [Hydroxyapatite. 2. Biochemistry]. AB - The Authors give a review of the literature of the biochemistry of hydroxyapatite especially about structure and formation of its crystals. PMID- 2561648 TI - [Hydroxyapatite. 3. Physiology of hydroxyapatite and bone calcification]. AB - The authors give a wide review of literature about the precipitation of hydroxyapatite, with particular attention to the latter studies about the collagene X and its leading role during this process. PMID- 2561649 TI - [Transient global amnesia as a manifestation of cerebellar metastasis]. PMID- 2561650 TI - The effects of alpha 2-adrenergic agonism with clonidine on the pulmonary and collateral resistances in the canine lung challenged with histamine. AB - In anesthetized dogs, we studied the effects of colonidine on the changes in pulmonary collateral resistance in response to histamine challenge. In the whole lung, an intravenous histamine increased the inspiratory (Ri), and expiratory (Re) resistance by 142 +/- 12%, and 229 +/- 30% over the control, respectively. Pretreatment with clonidine (0.05 mg/kg, iv) significantly attenuated the increases to 76 +/- 16% (Ri), and 114 +/- 22% (Re) (p less than 0.05). However, the responses to histamine were restored after an addition of yohimbine (0.15 mg/kg, iv) before clonidine. A bronchoscope wedge technique was used to measure the collateral resistance (Rcs) in the lung periphery. Histamine was administered in aerosolized form to a wedged sublobar segment. The local histamine challenge significantly increased Rcs. Pretreatment with clonidine did not discernibly affect the histamine-induced increase in Rcs. On the other hand, pretreatment with isoproterenol (0.02 mg/kg, iv) remarkably attenuated the Rcs response to histamine. The results indicate that alpha 2-adrenergic agonism with clonidine inhibits the bronchoconstriction induced by histamine. alpha 2- antagonism with yohimbine abolishes the antiasthmatic effect of clonidine. In the lung periphery, the Rcs response to histamine is not altered by alpha 2, but by beta 2 adrenergic agonism with isoproterenol. PMID- 2561651 TI - Evidence for the coupling of muscarinic M1 receptors to polyphosphoinositide turnover in rat cortical synaptosomes. AB - The linking event between the activation of muscarinic M1 receptors and the stimulation of polyphosphoinositide (PPI) turnover was studied in rat brain synaptosomes from the muscarinic M1 enriched cerebrum and M2 enriched cerebellum. The muscarinic M1 selective antagonists, pirenzepine and trihexyphenidyl, and M2 selective antagonist AF-DX-116 were chosen to displace the non-selective labeled cholinergic ligand, 3H-QNB, binding to these two regions of the brain synaptosomes. The IC50 values obtained for these agents revealed a typical characterization of the receptor subtypes of these two regions as they are. Carbachol-induced a stimulation of the PPI turnover cycle: namely, a decrease in 32Pi incorporation into phosphatidylinositol-4,5-bisphosphate (TPI) and phosphatidylinositol-4-phosphate (DPI), and an increase in this incorporation into phosphatidylinositol (PI) and phosphatidic acid (PA) in rat brain synaptosomes from the cerebrum. However, this event was only barely detectable in the synaptosomes from the cerebellum. The IC50 values obtained for these antagonists to block the carbachol-induced PPI turnover cycle in the synaptosomes from the cerebrum were close to the values obtained for the displacement of 3H QNB binding to the same preparation, and were far away from those values obtained in the synaptosomes from the cerebellum. Our results suggest that there is evidence to support the view that muscarinic M1 receptors are coupled to the PPI turnover event in rat cortical synaptosomes. PMID- 2561652 TI - Nutrient reabsorption of intestine in vitro: effects of ethanol and caffeine. AB - We studied the effect of ethanol and caffeine on the intestinal reabsorption (jejunum from SD rats) of glucose (Glu) and amino acids. Since most of the studies on the effect of ethanol utilized high concentration, we first characterized the effect of 8% (approximately 1.4 M) ethanol on the activity of Na(+)-coupled nutrient transport. Consistent with previous reports, ethanol (greater than 1 M) was found to inhibit the uptake rates of glucose and its non metabolizable analogue 3-O-methyl-glucose (3-OMG) by 30%, while leucine (Leu) uptake was inhibited by 60%. Phloridzin, a specific inhibitor for Na(+)-coupled sugar transport, at 1 mM concentration could inhibit Glu and 3-OMG uptake by more than 60% without affecting Leu uptake. We then compared the effects of various concentrations of ethanol on about 20 intestinal segments taken from the same animal. We consistently observed transport inhibition at high concentration of ethanol but at low concentrations (up to 200 mM), there was no consistent effect, while phloridzin or low-Na media (86% of Na replaced by choline) significantly reduced the rate of nutrient uptake in the same experiment. Thus, it appeared that low concentrations of ethanol had no significant effect on Na(+)-coupled nutrient uptake. We also determined the effect of caffeine on intestinal 3-OMG uptake. At concentration of 0.05 mM, caffeine inhibited 3-OMG uptake by about 15% (p less than 0.05). The level of inhibition was not significantly different at 0.5 mM, but a slightly higher level of inhibition (20%) was reached at 5 mM. The action of caffeine could be mimicked by dibutyryl cAMP (1 mM). PMID- 2561653 TI - [Fabry's disease: kidney insufficiency in heterozygous patient]. AB - We report a rare heterozygous status for Fabry's gene with severe kidney involvement and normal alpha-galactosidase A activity, together with the intrafamilial variations in the clinical expression of the disease. The random X inactivation hypothesis seems to explain such a variable expression of the alpha galactosidase gene in our cases. PMID- 2561654 TI - Carcinoid tumour of bronchus--masquerading as carcinoma. PMID- 2561655 TI - Granular cell myoblastoma of vastus lateralis muscle. PMID- 2561656 TI - [Air polishing instruments. Review of literature]. AB - The popularity of polishing air devices (AP) is rising with the increased distribution of commercial units in both United States and European dental offices and clinics. The instrument's efficiency and effectiveness in stain removal has been demonstrated with minimal impact on soft tissue trauma and abrasion. This article reviews the literature on AP. Future research is indicated to explore surgical clinical applications of airpowder polishing as well as more detailed information regarding the biological basis for its use. PMID- 2561657 TI - Infantile spasms--a clinical perspective. PMID- 2561659 TI - [Infectious agents which contaminate the upper airodigestive tract]. PMID- 2561658 TI - [Superoxide anion production and adhesiveness of neutrophils in obese patients]. AB - In 15 subjects with simple obesity and 10 patients with obesity associated with type II diabetes neutrophil adhesiveness and the rate of resting and stimulated production of superoxide anions (O2-) by these cells were assessed. High values were demonstrated of neutrophil adhesiveness suspended in autologous plasma in both groups of patients. The value of O2- production by resting cells was significantly raised, particularly in cases of simple obesity. Stimulated production of superoxides by neutrophils was approaching the value noted in the control group. The obtained results may suggest participation of these cells in the development of atherosclerotic changes. PMID- 2561660 TI - DNA restriction enzyme analysis of a bovine herpesvirus 1 strain isolated from encephalitis in Hungary. AB - The DNA of a bovine herpesvirus 1 (BHV-1) strain isolated from calf encephalitis in Hungary was analysed with restriction enzymes. The cleavage pattern of the encephalitis strain Na/67 differed from those of all the other Hungarian BHV-1 isolates investigated so far. The EcoRI and HindIII cleavage patterns of virus strain Na/67 were found to be similar to the patterns of two other encephalitis strains (N569 and A663 from Australia and Argentina, respectively) characterized earlier. Strain Na/67 is the first isolate in Europe which showed the restriction enzyme pattern of BHV-1.3 previously supposed to be characteristic of encephalitis strains. PMID- 2561661 TI - Molecular cloning of DNA from a bovine herpesvirus 1 strain isolated in Hungary. AB - Molecular cloning of the HindIII fragments of bovine herpesvirus 1 (BHV-1) strain HB144, isolated from infectious bovine rhinotracheitis (IBR) in Hungary, and of an infectious pustular vulvovaginitis (IPV) reference strain (K22) is reported. So far 52% of the IBR viral genome and 28% of the IPV viral genome have been cloned. The analysis of differences between the strains is currently in progress. PMID- 2561662 TI - The conserved open reading frame overlapping the thymidine kinase gene of different herpesviruses exists also in bovine herpesvirus 1. AB - Similar open reading frames (ORF) overlapping the thymidine kinase (TK) gene on the opposite strand or being close to it have been identified in 8 herpesviruses (Jacobson et al., 1989). Study of the DNA sequence of BHV-1 TK gene published by Mittal and Field (1989) and Kit and Kit (1986) revealed the existence of a similar ORF in this virus genome, too. PMID- 2561663 TI - An evaluation of four serological tests for the detection of antibodies to bovine parainfluenza virus type 3. AB - Haemagglutination-inhibition (HI), virus neutralization (VN), haemolysis inhibition (HLI-1, HLI-2) tests, and two new haemofusion-inhibition (HFI-1, HFI 2) tests, developed by us, were tested for the detection of antibodies to bovine parainfluenza virus type 3 (BPIV-3) in sera of 45 steers randomly selected from a herd naturally infected by BPIV-3. Twelve seronegative animals were then vaccinated with formalin-inactivated vaccine and 15 days later tested by the above-mentioned assays. Linear regression analysis revealed positive correlations between HI, VN, HLI-1 and HFI-1 antibody titres, which confirm the specificity of the HFI-1 test. The HLI-1 and HFI-1 assays proved to be less sensitive than the HI and VN ones, while HLI-2 and HFI-2 tests failed to detect any antibody to BPIV 3. PMID- 2561664 TI - [Experimental evaluation of two different diamond rotary instruments in finishing flat levels]. PMID- 2561665 TI - Chemotherapeutic mouthrinses as adjuncts in implant dentistry. PMID- 2561666 TI - [Molecular mechanism of action of nuclear hormonal receptors]. PMID- 2561667 TI - [Fructose-2,6-diphosphate and glycolysis of tumor cells]. AB - Tumour and proliferative cells maintain a high glycolytic rate even under aerobic conditions. The discovery of fructose-2,6-bisphosphate, a potent stimulator of glycolysis, has prompted a re-investigation of this phenomenon. Rat hepatoma cells and fibroblasts stimulated by mitogens or transformed by the Rous sarcoma virus, were used as models. The results indicate that the stimulation of glycolysis induced by these agents can be explained by an increase in the concentration of fructose-2,6-bisphosphate and in the activity of the enzyme synthesizing it. PMID- 2561668 TI - [AIDS and sleep disorders: effect of gp120 on cerebral glucose metabolism]. AB - The neurological complications associated with infection by the AIDS virus, HIV, occurs at an early stage of the disease and often indicate a poor prognosis. A dementia, known as AIDS Dementia Complex, is the most common feature observed, and is found in a majority of patients. The effects of gp120, the external protein envelope of HIV, on cerebral glucose utilization were studied in rats. Intracerebroventricular injection of gp120 significantly reduced glucose utilization in the lateral habenula and the suprachiasmatic nucleus, two regions rich in receptors for Vasoactive Intestinal Peptide (VIP) and the whole brain metabolism showed a significant decrease. The findings suggest that gp120 may alter neuronal function, thereby contributing to sequelae of HIV infection of the brain, and that attachment of HIV particles may involve, for a part, VIP receptors. PMID- 2561669 TI - Salivary gland tumors. AB - Although the incidence of salivary gland tumors in the general population is relatively low, these neoplasms account for a significant proportion of tumors of the oral and perioral regions and are therefore of considerable interest to the dentist. Malignant salivary gland tumors comprise the second most common type of oral cancer, although squamous cell carcinomas arising from the surface mucosa are nine times more common. Salivary gland tumors occur in both the major (parotid, submandibular, and sublingual) and minor (accessory) salivary glands of the oropharynx. This article will discuss various types of salivary gland tumors and treatment options. PMID- 2561670 TI - [Experiments on abrasion and wear of dental diamond instruments]. AB - In spite of identical ISO standards for shape, grain size, and largest head diameter of diamond abrasive instruments there are major differences in the machining performance among the various makes. These differences are largely due to variations in shape and diamond partical size. Diamond burs from different manufacturers of the same visual shape and particle size do not show any significant differences in performance. However, not only the abrasive performance but to a greater extent the design of the preparation margin determine the selection of a bur. It would be desirable that manufacturers be more specific about the particle size used and the shape of the bur, since the ISO standard does not include sufficiently accurate data about diamond particle size and shape design. The life of a diamond instrument is limited by the wear of its tip. Here the diamond layer wears off faster than on the rest of the instrument. This calls for early replacement of the instrument in clinical use even if the shaft still might be functionable. PMID- 2561671 TI - [Augmentation of mandibular bone with hydroxylapatite--follow-up]. AB - A group of 31 patients whose mandibles had been augmented surgically with various methods using hydroxylapatite granules between 1984 and 1987, were followed-up to rate their subjective and objective masticatory function. The subjective assessment of the treatment result is comparable to the opinion given about vestibulo- and floor of the mouth-plasty. Masticatory function determined according to the method of fractionated sieving of the chewed food did not show any significant differences from the function measured in complete denture patients with strong alveolar bones. The need for regular follow-up is emphasized. PMID- 2561672 TI - [Augmentation of alveolar process with hydroxylapatite--clinical orthodontic experience]. AB - 23 Patients were followed up clinically and by X-ray. In this cohort eight maxillary and 19 mandibular alveolar bone augmentations with hydroxylapatite granules mostly in a subperiosteal tunnelling procedure had been done. In most cases prothetic treatment was administered in our hospital. The dentures had been in place for a little over three years. Over this period of time there was hardly any change in the shape of the augmented ridges, denture retention, and vertical dimension. Drawbacks of the procedure became evident as reduced load resistance of oral mucosa during mastication and recurring impingement spots. In some cases, particularly when there was little vertical bone height, pain occurred in the area of the mental nerve. PMID- 2561673 TI - Loaded hydroxylapatite-coated and grit-blasted titanium implants in dogs. AB - This study evaluated the response of canine mandibular bone to loaded hydroxylapatite-coated (HAC) and grit-blasted titanium (GT) endosseous dental implants. Four dogs were partially edentulated in the maxilla and mandible. Two implants supporting freestanding prostheses were placed in each quadrant. Following 1 and 10 months of loading, the implants were evaluated. Soft-tissue pocket depths were not statistically different between the HAC and GT implants. Crestal bone loss was not significantly different between the two implants. However, the HAC implants had a statistically significant greater amount of bone apposed to their axial and apical surfaces compared to the GT implants. PMID- 2561674 TI - Modulation of calcitonin secretion by modification of calcium channels? AB - Voltage-dependent calcium channels (VDCC) regulating Ca++ influx through the cellular plasma membrane play a major role in the Ca(++)-induced calcitonin (CT) secretion. Using rat C-cells (rMTC 6-23 cell line), we have studied the effect of repetitive stimulation by either Ca++ (2 mM) or glucagon (10 microM) or epinephrine (10 microM) on CT secretion. Following a Ca(++)-induced initial rise, CT release declined to basal levels after about four hours despite high Ca++; addition of 10 microM glucagon to the "Ca++ desensitized C-cells" yielded the normal stimulatory effect of glucagon on CT release. Repetitive stimulation with glucagon showed a constant stimulatory action over an eight-hour period. In contrast, repetitive stimulation with 10 microM epinephrine caused an initial rise followed by a gradual decline of CT release over six hours. The observed desensitization of Ca(++)-induced CT secretion may be due to a modification of VDCC in C-cells. Whether or not the desensitization of epinephrine-induced CT release occurs independently of the regulation of VDCC remains unclear. PMID- 2561675 TI - Ethanol induced EMG changes and muscle force in a patient with polyneuropathy. AB - A patient with chronic relapsing polyneuropathy noticed that when he took some alcohol in the evening his muscle force increased substantially during the next day. To measure this beneficial effect of ethanol we measured the force of hand grip, conduction velocities and distal latencies of ulnar and median nerves, and the power spectra of EMG during maximal and submaximal hand grip before and after alcohol ingestion. A 50% increase in the muscle force was documented 24 hours after the alcohol administration, with a concomitant increase of high frequency components in the power spectrum of EMG. Slight decreases in distal latencies were also measured. PMID- 2561676 TI - [Anticalculus dentifrices. A new era in preventive dentistry?]. AB - Anticalculus toothpastes are available on the market. Toothpastes with Zinc compounds interfere mainly with the plaque formation. Dentifrices containing pyrophosphate give the highest reductions in calculus formation. The inclusion of pyrophosphate in a fluoride containing dentifrice did not interfere with the cariostatic action of fluoride on tooth enamel. The influence of anticalculus dentifrices on root caries and root hypersensitivity are, so far, not documented. Fundamentally, a lot of doubt still exists about the precise role of supragingival calculus formation in the onset of periodontal disease. PMID- 2561677 TI - [Preserving alveolar bone with submucosal hydroxyapatite]. AB - In this article the use of hydroxyapatite granules to prevent alveolar bone loss after extraction of teeth is described. Twenty-six alveoli were treated in the mandible and the maxilla. The used method resulted in a delayed healing of the wound and loss of a part of the implants. On the account of the results it is recommended to close the extraction wound completely when using hydroxyapatite granules and to limit the indication to the mandible. PMID- 2561678 TI - [Augmentation of the atrophic mandible with particulate hydroxyapatite]. AB - Experience in handling particulate HA granules in the augmentation of the atrophic mandible is described. Different methods like the use of fibrin glue and the subperiosteal tissue expander in relation with the type of atrophy are discussed. PMID- 2561679 TI - [Application of hydroxyapatite as an implant in oral surgery: an overview]. AB - An overview is presented of the clinical application of hydroxyapatite granulate as an implant material in atrophic, edentulous parts of the jaw. PMID- 2561680 TI - [Implants with plasma spray coatings of hydroxyapatite: an experimental study in alveolar bone]. AB - Sintered hydroxyapatite ceramic can be coated to a titanium core by means of a high temperature plasma spray procedure. The biological response of alveolar bone to an implant with such a coating of hydroxyapatite was evaluated in an animal experimental study. The results indicated that the plasma spray coating gave a very strong and direct bonding to cortical alveolar bone. The shear strength between bone and ceramic was greater than the shear strength between ceramic and titanium. From a radiological, macroscopic and microscopic point of view the conclusion could be drawn that the biological properties of a plasma spray coating of hydroxyapatite are the same as the properties of bulk hydroxyapatite ceramic. The outermost layer of the hydroxyapatite coating appeared to be unstable under the experimental conditions. PMID- 2561681 TI - Erythrocyte sodium pump activity in bipolar affective disorder and other psychiatric disorders. AB - Erythrocyte ouabain-inhibitable sodium pump activity, a measure of NaK-ATPase activity, was studied in 6 diagnostic groups of psychiatric subjects: bipolar affective disorder, unipolar depressive disorder, neurotic depression, chronic alcohol abuse, schizoaffective disorder, and schizophrenia, and in sex- and age matched normal controls. In the bipolar manic-depressive group, which was restricted to lithium-free subjects, values for sodium pump activity were significantly lower than in the controls (-11.4%, n = 53, p less than 0.001); subgrouping of the bipolar group by sex or age showed a significantly lower sodium pump activity in each of the groups. In the unipolar depressive group, values for sodium pump activity were significantly higher than in the controls (+13.7%, n = 12, p less than 0.01). The difference in direction of changed sodium pump activity between the bipolar and the unipolar groups was also observed in the values for subgroups of subjects in the two categories who were in a depressed state at the time the blood sample was taken. In the chronic alcohol abuse group, values for sodium pump activity were significantly higher than those for the control group (+13.5%, n = 20, p less than 0.05). In the neurotic depression (n = 24), schizoaffective (n = 12), and schizophrenia (n = 35) groups, there were no significant differences in sodium pump activity between the group of psychiatric subjects and their matched controls. These observations indicate that there is a trait-dependent deficiency of NaK-ATPase activity in bipolar affective disorder. PMID- 2561682 TI - Deficient erythrocyte NaK-ATPase activity in different affective states in bipolar affective disorder and normalization by lithium therapy. AB - This report expands on previously presented evidence for a trait-dependent deficiency of erythrocyte sodium pump activity in bipolar affective disorder. Several parameters of erythrocyte NaK-ATPase activity in different affective states and the effects of lithium therapy were examined. In lithium-free bipolar affective disorder patients, the mean percent differences from the individual sex and age-matched controls for erythrocyte sodium pump activity were: manic + hypomanic group, -21.5%, n = 16, p less than 0.02; depressed group, -12.4%, n = 14, p less than 0.02; euthymic group, -6.9%, n = 18, p less than 0.10. Ouabain sensitive potassium ion uptake was less than the controls in the manic group ( 25.4%, n = 3, p less than 0.02), and in the combined affectively ill group, manic + depressed (-23.4%, n = 7, p less than 0.02). Cell ouabain binding was less than the controls in the manic group (-19.0%, n = 6, p less than 0.05). NaK-ATPase activity in washed erythrocyte membranes (ghosts) was significantly lower than the controls in the manic group (-14.5%, n = 4, p less than 0.02), and the mean value for the whole group (manic + depressed + euthymic) was lower than the controls (-11.8%, n = 18, p less than 0.05). Values for ouabain binding in ghosts from the bipolar subjects were not significantly different from the matched controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561683 TI - Platelet [3H]-imipramine binding according to DSM-III subtypes of depression. AB - The question of the previously reported changes in the density of high-affinity binding sites for [3H]-imipramine (IMI) in platelets from depressed patients was reexamined among the different diagnostic subtypes of depression according to the DSM-III classification and taking into account the possible influence of the low affinity binding site. Using a least-square computer-assisted analysis, a precise determination of the [3H]-IMI binding parameters exclusively in relationship to the high-affinity site was performed in 46 untreated depressed patients and compared to 35 healthy controls. The results revealed a clear and highly significant 22% decrease in the maximal density (Bmax) of [3H]-IMI binding in all of the depressed patients compared to controls with no change in affinity values. Considering the diagnostic subgroups, we found that all the bipolar patients, the depressed as well as the euthymic or manic ones, had very low Bmax values and that some of them also exhibited an unusual low-affinity binding. Mean Bmax of the unipolar and dysthymic patients significantly decreased when compared to controls, although the Bmax values showed a large variability. Only dysthymic patients presented Bmax values which were significantly associated to symptom severity as assessed by the Hamilton Depression Rating Scale scores. Our results confirm the lower density of platelet [3H]-IMI binding in affective disorders, particularly in bipolar patients, and also suggest that this biological parameter is a trait marker in bipolar depression and a state marker in dysthmic disorder. PMID- 2561684 TI - Central nervous system in AIDS--an update. PMID- 2561685 TI - [Cartography of fluoride in the Ivory Coast (partial study)]. AB - The authors report on a study done in the Cote d'Ivoire, about the amount of fluoride in drinking water. The cartography of fluoride shows amounts lower than 0.7 ppm. This small amount of fluoride partially explains the increase in the number of dental caries in the Cote d'Ivoire. PMID- 2561686 TI - [Caries of the first permanent molar in Ivory Coast children]. AB - The authors study dental decay in the first permanent molar among school children in Dimbokro (Cote d'Ivoire). The study, carried out on 1050 children aged between 7 to 16, revealed a low CAO index but a frequency index greater than 50%. PMID- 2561687 TI - [Craniofacial anatomo-morphological profile of black African Ivory Coast children]. AB - An African Negroid ivorian child's facial, cranial, morphological and anatomical profile research The cephalometric study of telerdiography negatives of profile carried out on a sample of 52 Ivorian children from 11 to 14 years old, has shown some particularities which characterize the face and skull architecture. These particular features differentiate the Ivorian from the west Africans and from the French. According to the authors it would be better to establish the Ivorian's specific norms and to consequently adapt the face and tooth orthopedie therapeuties. PMID- 2561688 TI - [Ivorian typology: study of the crown dimensions of the first permanent molar]. AB - A dental morphology study is of great importance because it allows norms to be established which are adapted to the Ivorian situation. After a study of the canal morphology, we became involved in a study of the canal dimensions of the first permanent molars amongst Ivorian children. As a part of the 260 mouldings taken from 67 boys and 63 girls, we measured the following diametres: the diametre masio-distal (O-M-D) taken between the edge of the mesial bone and the edge of the distal bone. the biggest vestibulo-lingual coronary diametre (OV-LC). the diametre of the vestibulo-lingual at the level of the occlusal table (OV-LC). The data collected allowed us to reach the following conclusions: the first secondary molar with a M-D diametre (10.64 mm) is more important than the M-D diametre of its homologeous upper (10.15). however there is no significant difference between the OV-LC diametre of the first maxillary molar and that of the first mandibular molar. concerning the M-D or V-L diametres, the difference between the left side of the maxillary as with the mandibular is negligble. It's worth noting that the first permanent molar is smaller for girls when compared to boys, as well as their M-D diametres compared to their V-L. A comparative study between the TALLEC and MARSEILLIER plates showed that: the first upper molar for Ivorian children has a M-D diametre practically identical to that of European children, however the V-L and OV-LC diametres are smaller. the first bottom molar slightly bigger in all M-D and V-L diametres in Ivorian children in comparison to European children. PMID- 2561689 TI - [Organization of dentistry in the Ivory Coast. Evaluation and perspectives]. AB - The author of this article was interested in the organization of odontology on the Cote d'Ivoire. First of all, he studied the different odontology pillars existing: their composition and their problems. Then he explained that in spite of the sanituary structures and the dental personal in office, the needs of the population in buccal and dental care would not be satisfied as long as the authorities would not modify the present organization of Ivoirian odontology. Finally, he produced some concrete propositions aimed at, in the long run on improving the buccal and dental state (health) of the collectivity. PMID- 2561690 TI - [The effect of thyroxine on cAMP-dependent protein phosphorylation and ion transport in the sarcolemma of the heart and skeletal muscles]. PMID- 2561691 TI - [Protein kinases and the photosynthetic activity of chloroplasts]. PMID- 2561692 TI - [Phosphoprotein phosphatases]. PMID- 2561693 TI - [Periodontal therapy as an integrated part of restorative dentistry]. PMID- 2561694 TI - [Geriatric dentistry. How older patients adaptation to dentures can be eased]. PMID- 2561695 TI - [Atypical facial pain--a disease picture for interdisciplinary diagnosis and therapy by the physician and dentist]. PMID- 2561696 TI - [Compliance problems in the after care of patients with periodontal diseases]. PMID- 2561697 TI - [Mechanical properties of newer alginates and the tensile strength of bonding in hydrocolloid-alginate combinations]. PMID- 2561698 TI - [Cavity margin preparation and marginal finishing of composite restorations in premolars. An in vitro study]. PMID- 2561699 TI - [Two-part ITI-hollow cylinder and hollow screw implant]. PMID- 2561700 TI - [Thick polished hydroxyapatite ceramic as dental implant material]. PMID- 2561701 TI - [New methods of caries prevention]. PMID- 2561702 TI - [Diagnosis and epidemiology of early periodontal disease in adolescents]. PMID- 2561703 TI - [Use of gingival sulcus secretion for diagnosis of periodontal diseases]. PMID- 2561704 TI - [Separation of smear layer of amorphous dentin with a 25% tannic acid solution]. PMID- 2561705 TI - [Caries diagnosis in teeth with fillings]. PMID- 2561706 TI - [Class V gold foil fillings: an in vitro study of margin quality]. PMID- 2561707 TI - [Caries prevention. Results of an intense prevention program on caries prevalence over a 10-year period]. PMID- 2561708 TI - [Seven years experience with a composite system for premolar region]. PMID- 2561709 TI - [Properties and behavior of palladium and non-precious metal alloys that can be fired]. PMID- 2561710 TI - [Impression accuracy in indirectly prepared root canal points]. PMID- 2561712 TI - Japanese encephalitis causing public health problems in southwest Asia. The SEAMEO-Tropmed technical meeting. Bangkok, Thailand, 13-15 June 1989. Proceedings. PMID- 2561711 TI - [Mandibular alveolar ridge augmentation with hydroxyapatite. Apropos of 2 cases operated at the Pulido Valente Hospital]. AB - The authors briefly review the factors of bone less edentulous, describe prosthodontic difficulties and mention surgical methods used to improve prosthetics retention. The introduction of Hydroxylapatite (HA) in the field of oromaxillofacial surgery has opened new and promising ways to correct the edentulous, both in the stability and easiness of surgical results. The subperiostal tunnelling is by far the most widely used it often needs the use of acrylic splints to avoid complications. An open mucosal flap technique although more delicate and requiring increased surgical time, seems to us useful concerning the extent of the results, less morbidity and lower cost to the hospital patient. PMID- 2561713 TI - Japanese encephalitis virus encephalitis: an overview. PMID- 2561714 TI - A review of Japanese-B virus encephalitis in Malaysia. AB - JE is neither classified as an entity in the Malaysian Medical records system nor is it a notifiable disease but is grouped under the broad umbrella of viral encephalitis. There is no centralised program by the Ministry of Health specially for JE surveillance and control. JE is endemic, occurs sporadically throughout the country all year round. Asymptomatic inapparent infections have been found to be more frequent than acute clinical encephalitis cases, judging from results of previous serosurveys (Pond et al., 1954). JE vaccination has never been tried in Malaysia. In a relative sense, JEV infection unlike dengue virus infection, does not appear to be much of a problem in Malaysia. Perhaps, the laboratory confirmed cases represent only a small proportion of the total hospitalised cases that actually occurred. The reasons may be that these cases could not be confirmed by laboratory tests due to improper timing or failure to obtain the second serum specimen, or failure to perform lumbar puncture on patient's refusal. Attempts to improve the case detection rate of JE in Malaysia should be made namely, by increasing clinical index of suspicion, instituting better specimen collection procedures and by adopting rapid diagnostic tests. PMID- 2561715 TI - A review of Japanese encephalitis cases in the Philippines (1972-1985). AB - Few studies have shown that JE does occur in the Philippines with the majority of the cases affecting the 1-10 year age group in places where rice fields abound. The morbidity rate is 15-17%, with a mortality rate of about 7-30%. PMID- 2561716 TI - Japanese encephalitis in Thailand. PMID- 2561717 TI - Japanese encephalitis in children in northern Thailand. AB - Fifty-nine children with Japanese encephalitis admitted in Maharaj Nakhon Chiang Mai Hospital since 1984-1985 were studied. The male to female ratio was 1.18:1. The age range was between 1 to 14 years old with 74% in the age range of 6-14 years. The symptoms included change of consciousness (100%), fever (96%), headache (76%), convulsions (59%) and vomiting (52%). The neurologic signs, namely positive meningeal signs (61%), hyperreflexia (61%), positive Babinski's sign (49%) hemiplegia (42%), papilledema (22%), and other cranial nerve palsies (23%) were seen. Abnormal respiration were found in 23% and 8% of cases had hypertension. Most children (81%) had blood leukocytosis with predominant neutrophils. The average CSF white blood cell count was 200 cells per mm. with lymphocytosis in 76 percent of the patients. The average CSF protein was higher than normal. Almost all cases had normal CSF sugar levels. The JEV antibody response, mostly primary type, Occurred in about 62 percent of cases. All children received symptomatic and supportive treatment, such as antipyretics, anticonvulsants, anticerebral edema agents, adequate respiration and nutrition and physical and occupational therapies. Associated complications were treated according to the individual's need. The mortality rate and neurological sequelae were found in 17% and 57% of cases respectively. Eighteen percent of the patients suffered severe neurological sequelae. The neurological sequelae included memory deficit (46%), mental retardation (42%), hemiplegia (34%), emotional and behavioral disturbance (24%), epilepsy (20%), motor aphasia (16%), cranial nerve palsies (16%), involuntary limb movement (8%) and blindness (2%). PMID- 2561718 TI - Molecular considerations for the laboratory diagnosis of Japanese encephalitis virus. PMID- 2561719 TI - Control of Japanese encephalitis vectors. PMID- 2561720 TI - Development of second generation JE vaccine with genetic engineering techniques. PMID- 2561721 TI - A test production of inactivated mouse brain JE vaccine in Thailand. PMID- 2561722 TI - A field trial of Japanese encephalitis vaccine produced in Thailand. PMID- 2561723 TI - [Hydroxyapatite use in dental practice. 4. Hydroxyapatite as restorative material in implant failures]. PMID- 2561724 TI - [Hydroxyapatite use in dental practice. 5. Combination uses of hydroxyapatite]. PMID- 2561725 TI - [Hydroxyapatite use in dental practice. 3. Use of HA in implantation]. PMID- 2561726 TI - [Various rare tumors of the lung]. AB - Carcinoids, mucoepidermoid as well as adenoid-cystic carcinomas represent a group of uncommon neoplasm of the lung. Based on their own experiences in between 1984 and 1988, including 6 cases (4 of which were carcinoids, one was a mucoepidermoid carcinoma and One an adenoid-cystic tumor), the authors have reviewed the existing literature on this subject and related the tumors of the long with those of the trachea, pointing out the different problems associated with the site of origin, the tumor spread, and the histological pattern of these malignancies. PMID- 2561727 TI - Reactivation of antibody genes in Epstein-Barr virus transformed human B cell lines. A preliminary report. AB - Three immortalised human B cell lines, which had either last their capacity to secrete specific antibody or secreted low levels of antibody were studied in an attempt to reactivate or enhance antibody synthesis. A variety of different stimuli known to cause activation, proliferation and differentiation of normal B cells were used including polyclonal activators and recombinant interferon. Four of them, (namely LPS, anti-IgM, IL2 and IL6) effectively increased specific antibody synthesis after three days of culture. These preliminary experiments show that the loss or decline in immunoglobulin production by immortalised human B cells is, at least in some cases, reversible. PMID- 2561728 TI - [Hydroxyapatite. 1. Chemical and physical structure]. AB - The authors give a review about the latter studies of physics and chemist structure of hydroxyapatite. PMID- 2561729 TI - Sjogren's syndrome. PMID- 2561730 TI - Aneurysmal ("angiomatoid") fibrous histiocytoma in a child. AB - A case of aneurysmal ("angiomatoid") fibrous histiocytoma (AFH) in a 12-year-old girl is presented with its unusual clinicopathologic features. The lesion had the full microscopic characteristics of AFH described in prior reports, but it also had some features that differed from the original description of the disorder, such as the involvement of subcutis, its occurrence in the scalp, and a documented history of minor trauma. The lesion clinically resembled the gross features of hemangioma. The experience in the present case raises the need for considering AFH as one of major differential diagnosis of nodular cutaneous tumors in children that simulate malignancy such as angiomatoid malignant fibrous histiocytoma and Kaposi's sarcoma. PMID- 2561731 TI - So-called sclerosing hemangioma of the lung--two cases report with ultrastructural study. AB - Sclerosing hemangiomas of the lung are benign neoplasms of uncertain histogenesis. We analysed two cases of sclerosing hemangiomas of the lung with histochemistry and electron microscopy. They had a variegated histologic appearance characterized by an admixture of solid, hemorrhagic, papillary and sclerotic lesions. Characteristic uniform round cells, unique to this tumor, were found within the stroma in all lesions. In the electron microscopic examination, we found Weibel-Palade bodies like small bodies in the tumor cells. We suspect hypothesis originating in the endothelial cell can not be completely excluded yet. Sclerosing hemangioma is a distinct clinicopathologic entity and should be distinguished from other benign neoplasms or inflammatory lesions of the lung. PMID- 2561732 TI - Comparison between electrophysiologic and morphologic changes in lead induced peripheral neuropathy in rats. AB - Compound nerve action potential (CNAP) of the mixed peripheral nerve is composed of A alpha beta, A delta, and C potentials. All components of CNAPs in the sciatic nerve were recorded by stimulating the tibial nerve of both control and lead-poisoned rats. Marked decrease of nerve conduction velocity and prolonged duration were found in A alpha beta and A delta fibers especially in large myelinated A alpha beta fibers. The amplitude decreased in A alpha beta potential, but the area did not change. In C potential produced by activation of unmyelinated fibers, nerve conduction velocity slightly decreased, but the amplitude and area did not significantly change. Pathologic correlates revealed prominent segmental demyelination with significant decrease of large myelinated fiber densities. Minimal axonal degeneration of unmyelinated fibers was present. We can conclude that electrophysiologic changes in the lead-poisoned rats correlate with pathologic changes in them. PMID- 2561733 TI - Increased ATP requirement for activity of and complex formation by DNA topoisomerase II from human leukemic CCRF-CEM cells selected for resistance to teniposide. AB - We showed previously that nuclear extracts from teniposide (VM-26)-resistant sublines of the human leukemic cell line, CCRF-CEM, exhibited decreased DNA topoisomerase II activity and ability to form drug-stabilized covalent protein DNA complexes (Danks et al., Biochemistry 27:8861-8869; 1988). In the present study, we found that nuclear extracts of these sublines (approximately 50- and approximately 140-fold resistant to VM-26) required 2 and 8 times more adenosine 5'-triphosphate (ATP), respectively, to achieve half-maximal stimulation of unknotting activity compared to extracts from the sensitive cells. When novobiocin, a competitive inhibitor of ATP binding to topoisomerase II, was included in the reaction, this ATP requirement increased 2.5- to 4-fold with the CEM cell extracts and 3.5- to 12-fold with the resistant cell extracts. ATP produced a dose-dependent increase in VM-26-stabilized topoisomerase II-DNA covalent complexes with nuclear extracts from all three cell lines. Extracts from resistant cells, however, formed 40-80% fewer complexes than those from sensitive cells. A similar decrease was seen with 4'-[(9-acridinyl)amino]methanesulphon-m anisidide, to which the cells are cross-resistant. With nuclear extracts from sensitive cells, the tetralithium salt of 5'-adenylylimidodiphosphate (AMP-PNP), a nonhydrolyzable analog of ATP, was as effective as ATP in promoting the formation of drug-stabilized enzyme-DNA complexes. With extracts from the resistant cell nuclei, however, AMP-PNP was about half as effective as ATP in promoting complex formation. Our results demonstrate that the effect of ATP on strand passing activity of and drug-stabilized complex formation by topoisomerase II is decreased in the nuclear extracts from the drug-resistant cells and suggest a possible basis for this form of drug resistance. PMID- 2561734 TI - Clonal stimulation or inhibition of human colon carcinomas and human renal carcinomas mediated by transforming growth factor-beta 1. AB - We examined various human carcinomas and cells populating a single human neoplasm to determine whether they exhibit a heterogeneous response to the effects of transforming growth factor-beta 1 (TGF-beta). Using recently established human colon carcinoma and renal cell carcinoma under defined in vitro conditions, we observed intertumoral and intratumoral heterogeneity and polarity of responses to TGF-beta (growth inhibition or stimulation) that did not correlate with the metastatic phenotype of the cancer cells as assessed in athymic nude mice. TGF beta mediated both cytostatic and cytolytic effects against sensitive tumor cells, and these responses were not related to the effects of TGF-beta on the cell-cycle traverse. The human colon carcinoma and renal cell carcinoma, however, exhibited differences in the expression of TGF-beta receptors. PMID- 2561735 TI - Bovine papillomavirus type I induces resistance to Ca+(+)-induced terminal differentiation in murine keratinocytes. AB - Bovine papillomavirus type 1 (BPV-1) was expressed in established mouse BALB/MK epidermal keratinocytes. In each of the transfected cell lines, DNA synthesis was stimulated by epidermal growth factor (EGF) and inhibited by type beta transforming growth factor to an extent similar to that in parental cells. In contrast, the BPV-1-transfectants were resistant to the induction of terminal differentiation by extracellular Ca(+)+. First, BPV transfectants continued to respond to EGF in the presence of Ca(+)+, whereas the growth of BALB/MK cells was arrested. The characteristics of EGF-binding were identical in the two cell lines. Second, the Ca+(+)-switch failed to induce the activation of epidermal transglutaminase in BPV-1-transfectants. Thus, BPV-1 caused mouse keratinocytes to become resistant to Ca+(+)-induced differentiation without otherwise affecting the control of cell growth. PMID- 2561736 TI - Rapid increase in the activity of DNA topoisomerase I, but not topoisomerase II, in HL-60 promyelocytic leukemia cells treated with a phorbol diester. AB - There is significant evidence to suggest that protein kinase C and DNA topoisomerases are functionally linked in signal transduction pathways. Much of this is based on the observation that phosphorylation of topoisomerase II by protein kinase C may lead to its activation in vitro and that inhibitors of topoisomerase II block phorbol diester-induced differentiation in HL-60 cells. In the present study, the activities of the DNA topoisomerases I and II have been quantitated to examine their regulation in phorbol diester-treated HL-60 cells undergoing differentiation. The activity of topoisomerase I increased rapidly after treatment with phorbol myristate acetate (PMA); it increased maximally (150% of control activity) at 3 hr post-treatment and remained elevated for at least 24 hr. Conversely, from the onset of exposure to PMA through 12 hr, there was no measurable alteration in topoisomerase II activity in PMA-treated cells. Moreover, there was a measurable decrease in topoisomerase II activity at the later time points, a result that occurred concomitantly with the loss of proliferative potential in differentiating HL-60 cells. Similar results were obtained when the activities of both enzymes were measured in nuclear extracts. The apparent increase in topoisomerase I activity was not due to an increase in the mass of the enzyme after PMA treatment, as measured by both western blotting and by the formation of camptothecin-dependent, topoisomerase I-DNA complexes. Taken together, these data suggest that the activities of the topoisomerases I and II may have been regulated independently in PMA-treated HL-60 cells, that the activity of topoisomerase II was not increased under conditions in which protein kinase C was activated in vivo, and that an increase in the activity of topoisomerase I may have had a role in the mechanism through which HL-60 cells underwent monocytic maturation in response to phorbol diesters. PMID- 2561737 TI - WEHI-3B D+ Y1 leukemia cells as a model system to assess the induction of differentiation in vivo. AB - An in vivo model system for assaying the induction of differentiation by therapeutic agents was developed using WEHI-3B D+ Y1 myelomonocytic leukemia cells in BALB/c mice, which exhibit characteristics analogous to those of human acute non-lymphocytic leukemia. An integrated copy of the non-mammalian 3' aminoglycoside phosphotransferase gene in WEHI-3B D+ Y1 cells permitted the unambiguous identification of these leukemia cells in vivo by in situ hybridization. The administration of aclacinomycin A to BALB/c mice bearing this leukemia produced an increase in survival time. In situ analyses of peritoneal cells obtained from anthracycline-treated animals supported the concept that the increase in life-span was due in part to the induced maturation of WEHI-3B D+ Y1 cells to more mature granulocytic elements. This action was accompanied by an order of magnitude decrease, relative to vehicle-treated animals, in the peritoneal leukemic cell burden. The schedule and dosage of the anthracycline antibiotic that produced the maximum prolongation of life had no effect on peripheral platelet levels or packed red cell volume. Elevated white blood cell levels were observed in leukemia-bearing mice; these increases were reversed in animals receiving 2 mg/kg of aclacinomycin A and reached control levels by day 8. Treatment with aclacinomycin A had no effect on body weight, or femoral or splenic cellularity. Increased numbers of granulocyte-macrophage colony forming units were observed in leukemia-bearing animals. Leukemia-bearing mice receiving aclacinomycin A decreased these stem cell levels to that of drug-treated non leukemia bearing animals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561739 TI - [A radiologic study of metastatic pulmonary tumor originating in the oral and maxillofacial regions]. AB - A retrospective study of 103 cases of metastatic pulmonary tumors originated from oral and maxillofacial regions was made radiologically as well as clinically. With regard to the site and pathologic type of the primary neoplasm, palatal tumor and adenoid cystic carcinoma was the most frequent ones respectively which metastasize to the lungs. Radiologic types of metastatic pulmonary tumor might be classified into five categories: solitary, multiple nodular, multiple mass, diffuse and miliary network. The relationship between prognosis and radiologic types, the growing rate and complications such as cavitation, atelectasis, pleural effusion, lymph node enlargement and destruction of ribs were briefly discussed. The authors were of the opinion that the duration from primary to metastatic and that from metastases to death in oral and maxillofacial malignant tumors are longer than those of other sites in human body. PMID- 2561738 TI - Expression of protein-associated DNA damage in the alkaline elution assay in the absence of enzymatic deproteinization. AB - In the alkaline elution assay, expression of protein-associated DNA damage induced by topoisomerase II antagonists is facilitated by proteinase K digestion of the drug-stabilized topoisomerase-II-DNA complex. In the absence of this enzymatic deproteinization step, drug-induced DNA strand breaks are masked by the binding of the topoisomerase-II-DNA complex to the synthetic filter from which DNA is eluted subsequent to alkaline denaturation. In this manuscript, we report that as the number of cells lysed on the filter is increased, binding of the topoisomerase-II-DNA complex to the filter is compromised, permitting expression of DNA damage in the absence of enzymatic deproteinization. PMID- 2561740 TI - [Periodontal disease and crevicular neutrophils. Role of superoxide radicals]. AB - One early biochemical process in PMN activation is the stimulation of the NADPH oxidase membrane enzyme, which produces a flux of superoxide radicals, directed towards the extracellular place. These radicals initiate bacterial destruction; however they can react against the periodontal tissue and lead to its destruction, either when too many of them are produced, or when they are not adequately neutralized by the antioxidant activity of the gingival fluid. Striking a proper balance between the activation state of crevicular PMN and the antioxidant activity of the gingival fluid may be a critical factor in determining whether PMN response to plaque bacteria is either protective or destructive for the parodontium. PMID- 2561741 TI - Minimal tooth preparation techniques for restorations with adhesive materials. AB - Dramatic progress over the past ten years in adhesive composite and glass ionomer materials has made possible innovative, minimal cavity designs. These designs, by conserving tooth tissue, maximize the strength of the remaining structure and facilitate the aesthetic preservation of the patient's own dentition. The author gives a critical review of some of the new techniques that have been proposed. PMID- 2561742 TI - Bone response to a matched modulus endosseous implant material. AB - The aim of this study was to assess bone response to a novel matched modulus material using more rigid titanium as a control. Implants subjected to different loading conditions were retrieved from rabbit tibiae after various time intervals, and new bone growth was assessed by histomorphometric methods. Block face microscopy was employed for histologic purposes. From a statistical analysis of the histomorphometric results, it could be concluded that titanium implants encouraged the greatest overall amount of bone formation around them when they were unloaded, when they were loaded immediately, and after a healing period. PMID- 2561743 TI - In vivo plaque formation on implant materials. AB - Plaque formation was studied in vivo by setting different implant materials on gingiva. The number of adhering viable bacteria depended on material surface properties 4 hours after setting, but not 48 hours after setting. The formation of pellicle-like thin layers and subsequent covering by lamellarly formed plaque were observed on the surfaces of all materials. Streptococcus species were predominant at the 4-hour setting time and anaerobes increased at the 48-hour setting time; this was common to all materials. The results indicate that surface properties of the implants influence early bacterial adherence, but do not influence bacterial flora or plaque maturation. PMID- 2561745 TI - Coralline porous hydroxyapatite as a bone graft substitute in orthognathic surgery: 24-month follow-up results. AB - A 24-month postsurgical follow-up evaluation was performed on 49 patients (20 males and 29 females; mean age 26 years [range 9 to 52 years]) who had undergone orthognathic surgery in which coralline porous block hydroxyapatite (PBHA) was used as a bone graft substitute. Followup ranged from 23 to 45 months (average 27 months). One surgeon implanted all 215 blocks: 189 blocks to the maxilla, 19 blocks to the mandible, and seven blocks to the midfacial region. Of the 215 implants, 135 were positioned directly adjacent to the maxillary sinus. Nine patients had postsurgical complications: three developed sinusitis, which was treated with antibiotics and decongestants, four had midpalatal implants exposed intraorally during surgery that later required removal, one had a slight displacement of a mandibular implant that has remained in place, and one had an interdental implant exposed that was recontoured and covered with tissue. At the 24-month followup or later, only one of the 49 patients had complications or symptoms associated months postsurgery. These results strongly support the use of coralline PBHA along with bone plate fixation to provide predictable stability in orthognathic surgery. PMID- 2561744 TI - Radiation leukemia virus and its effect on H-2 gene expression. AB - In this report we demonstrate that lowered expression of the H-2 antigens on RadLV-induced tumour cells is a result of depressed levels of stable mRNA in these cells. Whether this observation is a result of lowered transcription or of mRNA instability is under investigation. In an effort to determine which viral sequences are essential for mediating both the H-2 regulatory function and the transforming function of RadLV, we have begun to assemble newly integrated proviral genomes from tumours. The restriction enzyme cleavage sites of four isolates are presented; these isolates differ substantially from RadLV genomes previously presented. One of these molecular clones is shown to encode a non defective B-tropic, ecotropic virus which when reinjected into resistant mouse strains can mediate the up-regulation of H-2Dd antigen expression. Finally, possible mechanisms of H-2 regulation are discussed. PMID- 2561746 TI - Chin augmentation with various alloplastic materials: a comparative study. AB - This retrospective study of 62 patients compared three alloplastic materials used in chin augmentation: Proplast I (PI), Proplast II (PII), and porous block hydroxyapatite (PBHA). There were 25 patients with PI implants (average follow-up of 44.8 months), 25 patients with PII implants (average follow-up of 32.8 months), and 12 patients with PBHA implants (average followup of 19.1 months). Average bone resorption was 1.25 mm (range 0.00 to 3.30 mm) beneath the PI and PII implants and 0.00 mm beneath the PBHA implants. Average posterior settling was 0.46 mm (range 0.00 to 2.00 mm) for the PI and PII implants and 0.03 mm (range 0.00 to 0.17 mm) for the PBHA implants. The percentage of soft tissue change at pagonion relative to the size of the implant was 83.0% (range 66.7% to 100.0%) for PI and PII and 86.8% (range 77.3% to 100.0%) for the PBHA implants. When PI or PII implants were used, younger patients had more resorption than older patients. Slightly more resorption was found with PI and PII implants when maxillary surgery was performed. The size of the PI or PII implants made no difference in the amount of bone resorption, although there was slightly less soft tissue change when larger implants were used. Males had slightly more bone resorption than did females. The bone resorption beneath the PI and PII implants occurred within the first 12 months postsurgery. It can be concluded from this study that PI and PII implants are acceptable materials for chin implants, but some resorption can be expected during the first 12 months postsurgery. The absence of detectable resorption and the excellent soft tissue change make PBHA an attractive alternative as a chin implant material, although it is technically more difficult to use. PMID- 2561747 TI - Coronal microleakage of permanent lingual access restorations in endodontically treated anterior teeth. AB - Forty-six intact extracted human anterior teeth were treated endodontically with laterally condensed gutta-percha and sealer. The teeth were restored with a base of either zinc phosphate or temporary stopping and a permanent restoration of either acid etched composite resin with GLUMA as the dentin bonding agent or with Ketac-Fil glass ionomer. The teeth were thermocycled, coated with nail varnish (except for the access), immersed in silver nitrate, developed, and sectioned longitudinally. The linear dye penetration was measured. All restorations permitted leakage into the base. All groups had specimens which leaked into the gutta-percha. There was a tendency for the glass ionomer/zinc phosphate group to leak least, but there were no statistically significant differences among the groups. PMID- 2561748 TI - Can we survive another change? PMID- 2561749 TI - [Cluster headache, Epstein-Barr and Burkitt's lymphoma virus infections]. AB - Thirteen patients affected by Burkitt's lymphoma were studied. Six out of them had cluster headache associated with Burkitt's lymphoma, and in this group we observed the absence of HLA-B14 antigen. The presence of HLA-B14 antigen in the remaining seven patients, confirm the hypothesis of a protective role of this antigen against cluster headache. PMID- 2561750 TI - Antithrombotic therapy in phlebopathies of lower limbs: a controlled study of low molecular weight heparin versus heparin calcium. AB - A study was made of 60 patients, 28 males and 32 females, with an average age of 62 years (range 52 - 81) suffering from phlebopathies of various etiology and pathogenesis in the lower limbs, treated with low molecular weight heparin (LMWH) or with standard heparin calcium. Posology was decided on the basis of the type of phlebopathy: the group treated with LMWH (30 patients) were administered one 7500, 15000 or 30000 AXaU vial/day sc; the comparison group were administered one or two 12500 IU vials/day sc of heparin calcium. Treatment lasted for 60 days. During treatments the symptoms and the clinical signs showed clear improvement in both groups, in a parallel and substantially analogous way. In all patients there was a discrete improvement in venous functioning, as shown by the drop in recumbent and erect venous pressure. Systemic tolerance was good. The only difference between the two treatments was represented by the local tolerance, which was markedly better in the group treated with LMWH. PMID- 2561751 TI - Detection of cytomegalovirus in paraffin-embedded lung tissue sections. AB - Attempts were made to detect cytomegalovirus (CMV) in paraffin-embedded lung tissue sections from five patients with malignant disease by the immunofluorescence and immunoperoxidase methods with application of monoclonal antibodies against CMV and by localization of DNA by the method of in situ hybridization using a biotinylated probe. In the immunofluorescence method, the monoclonal antibody 1C6 stained a large number of CMV-infected cells. However, nonspecific staining of the bronchoepithelial cells was a disadvantage of this method. In the immunoperoxidase staining, the monoclonal antibodies E. 13 and CCH2 gave highly sensitive and specific results. These monoclonal antibodies stained the nuclei of the infected cells even in the absence of viral inclusions. In the viral DNA probe technique, three specimens were found to contain hybridizing cells. The DNA probe detected not only cytopathic cells but also normal-appearing infected cells. The number of cells stained by the DNA probe was much smaller than that stained by the immunoperoxidase method. The author concludes that the immunoperoxidase technique using monoclonal antibodies against CMV is useful for the diagnosis of CMV infection in the paraffin-embedded lung tissue. Localization of DNA by the method of in situ hybridization is not as useful as the immunoperoxidase method because of lower sensitivity. PMID- 2561752 TI - [Paraneoplastic endocrine syndromes]. PMID- 2561753 TI - [IgG anti HSV I antibodies in saliva in a drug dependent population]. AB - It is known the presence of receptors for the morphine products on the Lymphocytes B. These receptors could have a modulator action on the production of antibodies in drug addicts. We have determined the existence of IgG anti-HSV I in saliva and serum in a heroin population. Our results show that here is not significant differences between the heroin population and the control group, when the antibodies are detected in serum. But significant differences do exist between the number of people with antibodies anti-HSV I in saliva. PMID- 2561754 TI - [Characterization of a cloned DNA fragment of rice chloroplast and location rbcL gene in the recombinant plasmid]. AB - The pure rice (Oryza sativa, subsp. Indica, cv. Zhenshan 97B) chloroplast DNA was digested by restriction enzyme BamHI and the resulting fragments were ligated to the BamHI site of pBR322. One recombinant plasmid which contains a 19.3kb insertel DNA fragment was isolated from the clone bank and was named pOSB1. A physical map of the recombinant plasmid was constructed by cleavage with ten restriction endonucleases, and the gene rbcL was located on the pOSB1. PMID- 2561755 TI - Three decades of chemotherapy for childhood cancer: from cure 'at any cost' to cure 'at least cost'. AB - The prognosis for childhood malignancy has improved substantially over the last 30 years. This has been principally due to the use of chemotherapy, centralization of care and treatment by standard protocols. With increasing numbers of children and adolescents cured of cancer there has been an increased awareness of the late effects of therapy. The recent recognition of disease specific prognostic factors has permitted the subclassification of tumours into good and bad risk. The aim for those malignancies associated with a good outcome is the reduction of late adverse effects, eg intellectual impairment in children with acute lymphoblastic leukemia receiving central nervous system irradiation. In those diseases which have a poor prognosis with present therapy, new innovative and intensive regimens are being investigated in randomized trials. Neuroblastoma is a chemosensitive tumour but there has been little success in translating response into improvements in long-term cure. In poor prognosis neuroblastoma conventional combination chemotherapy, high dose therapy with autologous bone marrow rescue as consolidation and more recently high dose, or high dose rapid schedule, chemotherapy as initial treatment have been sequentially evaluated. In paediatric brain tumours only now are well designed phase II studies being undertaken and problems associated with the classification and evaluation of response to therapy are being addressed. In the past 30 years much of the success with childhood cancer has been largely empirical and the hope for the future is that there will be a more scientific approach to the chemotherapy of childhood malignancy. PMID- 2561756 TI - The impact of chemotherapy in germ cell tumours. AB - It is widely appreciated that the application of chemotherapy in the treatment of germ cell tumours exemplifies the best results to be expected from this approach in solid tumours, since the majority of patients treated are now cured. Against a background of steadily rising incidence of testicular cancer, the impact of this highly effective treatment can be measured using national statistics, which for most countries in the developed world indicate a corresponding fall in mortality over the past 15 years. For some countries, however, no such improvement has taken place, presumably reflecting inadequate facilities in those countries for securing appropriate treatment, particularly cisplatin. In most specialist centres, current clinical studies are aimed at reducing the toxicity of treatment for the majority of patients cured of their disease. The other remaining challenge in management is to improve the results of treatment in the small minority of patients for whom chemotherapy is still not curative. More intensive schedules are being tested, and randomized trials to test these approaches are recommended. PMID- 2561757 TI - Effects of various prophylactic treatments on titanium, sapphire, and hydroxyapatite-coated implants: an SEM study. PMID- 2561758 TI - Hydroxyapatite-augmented sites as receptors for replacement implants. AB - Implant failures can arise from a variety of conditions. Damage to the tissue structures at surgical placement, inadvertent stress distribution to the marginal bone, unfavorable arch relationships between the maxilla and mandible, occlusal trauma, inadequate prosthetic design, and improper oral hygiene are but a few of the problems that can result in ultimate damage and loss. The use of hydroxyapatite to restore bone contour and health can allow for replacement of a dental implant reconstruction. This paper will describe the use of porous hydroxyapatite to create proper bone morphology for the placement of IMZ osteointegrated dental implants in an edentulous maxilla and a mandibular distal edentulous area. Both cases are followed from the removal of previously placed blade-type implants to the reconstruction by use of the IMZ osteointegrated system within the hydroxyapatite-contoured ridges. PMID- 2561759 TI - Histological response and clinical evaluation of heterograft and allograft materials in the elevation of the maxillary sinus for the preparation of endosteal dental implant sites. Simultaneous sinus elevation and root form implantation: an eight-month autopsy report. PMID- 2561760 TI - Scanning electron microscopic study of cell attachment to biodegradable polymer implants. AB - The biodegradable polymers--polylactic acid (PLA) and polyglycolic acid (PGA)- are currently being studied as carriers for bioactive bone regeneration compounds. The inclusion of osteo-inductive substances in poly-(DL, lactide-co glycolide) copolymer alloplastic implants has been shown to enhance the repair of osseous defects. The purpose of this study was to examine, by SEM, the attachment relationship of biodegradable polymer implants to cells and tissue matrix. Three groups of copolymer implants were studied: (1) plain 50:50 PLA-PGA copolymer, (2) PLA-PGA copolymer with hydroxyapatite (HA), and (3) PLA-PGA copolymer with autolyzed, antigen-extracted (AA) bone particles. Polymer discs were surgically implanted into the pectoralis muscles of rats. At seven, 14, and 21 days post implantation, the baskets were removed and the contents prepared for SEM. Results showed that at one week, implants were coated primarily with red and white blood cells in a fibrinoid clot. Degradation of the polymers was evidenced by irregular enlarging of polymer surface pores. At two and three weeks, polymers became lobular and then fibrinoid as degradation progressed. Inflammatory cell and red blood cell adhesions were increasingly replaced by fibroblasts and collagen matrix deposition. As polymer degradation progressed, AA and HA particles were exposed; however, the lack of cell or tissue adhesion in these areas suggests that degradation may be more influenced by the fluid environment than by direct cell attachment. Furthermore, degradation may inhibit direct cell attachment. PMID- 2561761 TI - Clinical and histological case study using resorbable hydroxylapatite for the repair of osseous defects prior to endosseous implant surgery. AB - A clinical case--a follow-up clinical and histological study--is presented in this paper, which describes the use of resorbable hydroxylapatite [OsteoGen (H.A. Resorb), Impladent, Inc., & GBD Marketing Group, Inc., Medical Division, Valley Stream, NY 11580] for the repair of a large mandibular lesion, followed by the placement of an endosseous dental implant six weeks after the graft placement. A comparison of the histological results of four-month and 14-month bone biopsies of resorbable hydroxylapatite grafts is included in this report. PMID- 2561762 TI - [Defects in receptors and post-receptor signal transduction]. PMID- 2561764 TI - [Receptor pharmacology and clinical application]. PMID- 2561763 TI - [Neuropsychiatric diseases and receptor abnormalities]. PMID- 2561765 TI - [Receptors of tumor cells]. PMID- 2561766 TI - [Experimental technics concerning receptor binding]. PMID- 2561767 TI - [Clinical tests related to receptors]. PMID- 2561768 TI - [Molecular biological information necessary for the study of receptors]. PMID- 2561769 TI - [A list of drugs acting through receptors]. PMID- 2561770 TI - [Description of receptors in relation to their target substances]. PMID- 2561771 TI - [Receptors. An outline]. PMID- 2561772 TI - [Description of receptors and related cells and tissues]. PMID- 2561773 TI - Identification and characterization of an avian reovirus isolated from black tailed gull (Larus crassirostris). AB - An isolated virus from the feces of nestlings of Black-tailed gulls living on the Kabu-Island, Hachinohe-city, Aomori-prefecture, was identified as an avian reovirus from its morphological, physicochemical and biological features. Serologically, the isolate has a 62% of relatedness to TS-17 strain, a prototype of avian reovirus in Japan. The isolate showed no significant virulences to one day-old SPF chickens, and showed low mortalities to chicken embryos, although it formed remarkable lesion onto chorioallantoic membrane. PMID- 2561774 TI - [Evolution of laboratory technics in the detection of herpes simplex virus infections]. AB - In this report we proved that all of three methods for laboratory diagnosis of HSV infections which we used, have approximately the same degree of sensibility. Even though there is no statistically significant differences among sensibility of chosen methods, we think that IF test by monoclonal antibodies is the most suitable method for the routine work, in consideration of the high sensibility, typical specificity and fast results. PMID- 2561775 TI - Bacteroides gingivalis vesicles mediate attachment of streptococci to serum coated hydroxyapatite. AB - Outer membrane vesicles purified from Bacteroides gingivalis culture supernatant bound to serum coated hydroxyapatite (SeHA). The immobilized vesicles served as receptors for a number of species of oral streptococci. The binding of Streptococcus sanguis 12 to SeHA was increased 10 times by the vesicles. Vesicle associated binding increased proportionally with an increase in the number of bound vesicles. Arginine and lactose both partially reduced binding of S. sanguis. Heating the vesicles destroyed their binding ability whereas heating S. sanguis reduced but did not eliminate vesicle-mediated binding. PMID- 2561776 TI - [Maxillofacial surgery for clefts]. AB - This paper makes a wide review of the pathology that affects clefts, where maxillofacial surgery plays a very important role in the rehabilitation of these patients. Here we will analyse the alveolar ridge repair orthodontic and orthognathic surgery of sequelae, and the use of halogenic grafts, bone integrated, for the filling of clefts. PMID- 2561777 TI - [Dental reimplantation using biological glues and hydroxyapatite. A clinical case]. PMID- 2561778 TI - Factors influencing the vascular action of dopamine in the canine mesenteric bed. AB - The effect of dopamine on the mesenteric arterial bed was investigated in dogs anaesthetized with pentobarbital sodium. The vascular responses to dopamine in a branch of the superior mesenteric artery supplying a segment of the small intestine were measured with a flowmeter probe and visualized by infrared telethermography or, in a separate series of dogs, were obtained by the direct determination of arterial resistance--changes during perfusion with constant flow. It was found that the mesenteric action of dopamine (given i.v. in the submaximal dose of 16 micrograms.kg-1.min-1 or intraarterially up to a dose of 40 micrograms.min-1) is primarily mediated via dopaminergic vasodilator and a alpha adrenergic vasoconstrictor receptors, the net result of this competition usually being moderate vasodilation under natural conditions. The contribution of beta adrenergic vasodilation to the mesenteric dopamine action is minimal as evidenced by beta-blockade with oxprenolol. By blocking the alpha-component with phentolamine (1.0 mg.kg-1) an almost threefold increase of the vasodilation was obtained in resistance. Because of the concomitant reversal of the systemic hypertensive dopamine action to hypotension, the net flow increase remained essentially unchanged. It was concluded that unless the degree of alpha stimulation is restrained and proper control of blood pressure is ensured, it is not possible to recommend that dopamine be used in the therapy of mesenteric vascular disorders. PMID- 2561779 TI - [Herpes labialis. Its diagnosis]. PMID- 2561780 TI - Investigation on the free radical producing effect of hematoporphyrin (a spin trapping study). AB - Direct evidence was given concerning the free radical production of hematoporphyrin (HP) and its derivative, during illumination, by ESR spectroscopy. A spin trapping measurement was carried out using PBN spin trap for determination of this radical(s). Three different spin-adducts were found. Two of them are likely to be connected to the reactive free radical production of HP. PMID- 2561781 TI - Spin-trapping studies with MTDO, a dihydroquinoline type radical scavenger. AB - The effect of MTDQ, a dihydroquinoline type free radical scavenger was studied in biological membranes, and model systems. Using spin-labelling and spin-trapping methods, it was that MTDQ induced a decrease of the mobility of spin labels attached to the thiol sites of membrane proteins, and it was a strong free radical scavenger in hydroxyl and superoxide anion free radical generating systems. The experimental results suggest that the presence of MTDQ in the cell membranes may improve the protection of membrane components against free radical attacks. PMID- 2561782 TI - Lipid lateral diffusion measurements in model membranes. AB - The rate of lipid lateral diffusion has been investigated by computer simulation of electron spin resonance (ESR) spectra of spin-labelled dimyristoyl phosphatidylcholine (DMPC) vesicles. An optimization method has been developed to fit the experimental spectra to the theoretical ones calculated from the modified Bloch-equations in order to determine frequencies of probe-probe collisions and the lipid lateral diffusion coefficients. The main results of this study are: (i) Due to the sensitivity of our method to the extent of the overlapping of hyperfine spectral lines it is possible to determine the spin exchange contribution to linebroadening. (ii) It is obvious from these computer analyses that over a wide range of temperatures well above the phase transition both static dipolar interaction and dynamic spin exchange make significant contributions to the linebroadening. (iii) Lipid lateral diffusion coefficient in DMPC bilayers at 36 degrees C was (2.3 +/- 0.2) x 10(-11) m2 s-1. PMID- 2561783 TI - Crystal morphology and decalcification patterns compared in rat and human enamel and synthetic hydroxyapatite. AB - The purpose of this investigation was to compare morphology and dissolution patterns by ultrastructural examination of rat and human enamel crystals as well as synthetic apatite crystals. Mature enamel crystals were of particular interest, since crystal maturation appears to be inhibited in amelogenesis imperfecta. Specimens were isolated from developing and mature rat incisor enamel. Rat enamel, mature human enamel, and synthetic apatite were thin sectioned without decalcification and examined by transmission electron microscopy. Some sections were exposed to acid, and selected synthetic apatite sections were further treated for removal of embedding plastic, followed by vacuum-shadow-coating with carbon. Results showed that cross-sections of rat, human, and synthetic crystals had a distortion in the flattened hexagonal outline in regions where the growth of one crystal impinged on another. Crystal dissolution occurred preferentially along the c-axis, producing a central defect or hole in the crystals. Preliminary studies with weak acid on mature human enamel indicate that the relatively soluble crystal core is quickly dissolved, while the outer shell remains intact over a much longer period of time. In the mature rat and human enamel, this crystal hole formation had a consistent dimension of approximately 10-nm thickness. The crystal hole dimension was the same size as crystals that are formed during the early secretory phase in rat amelogenesis. Acid-treated synthetic apatite also showed dissolution of the crystal core along the c-axis, but dimensions of the hole were not consistent.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561784 TI - Characteristics of a 5-hydroxytryptamine-sensitive adenylate cyclase in intact and intracellularly perfused squid axons. AB - 1. Cyclic AMP metabolism was studied in intact and intracellularly perfused axons. 2. Cyclic AMP content of intact axons lay within the range 10-100 nmol kg 1 axoplasm. This was increased by exposure to caffeine (2-fold) and to 5-HT (15 fold). The caffeine-sensitive cyclic AMP increase was 30-fold larger in the presence of 5-HT. 3. A reduction in sodium concentration from the sea water bathing intact axons attenuated the 5-HT-evoked increase in cyclic AMP content, but had little effect on resting cyclic AMP. This effect was partially reversed by exclusion of external calcium, and suggests that free calcium plays a role in cyclic AMP homeostasis. 4. Prolonged exposure of intact axons to 5-HT (up to 3 h) led to apparent desensitization of the cyclic AMP response. 5. Intracellular perfusion can be used as a method to study adenylate cyclase in a single axon, simply by measuring the cyclic AMP content of the emerging perfusate. 6. Intracellular perfusion revealed micromolar requirements for internal GTP (K0.5 approximately 1-10 microM) and external 5-HT (K0.5 approximately 1-10 microM); a detailed investigation of this observation was limited due to the progressive loss of 5-HT-evoked adenylate cyclase activity with time. This slow loss was not seen during Gpp(NH)p (guanylylimidodiphosphate), NaF or forskolin activation of cyclase activity. 7. In perfused axons, an increase in intracellular calcium stimulated cyclase activated by 100 microM-forskolin but inhibited cyclase activated by 500 microM-Gpp(NH)p or 10 mM-NaF. A reduction in intracellular magnesium from 10 to 4-5 mM attenuated the effects of 5-HT-evoked cyclase activity. 8. Study of the perfused axon allows characterization of the intracellular requirements of a plasmalemmal transduction system which activates adenylate cyclase, whilst maintaining ionic asymmetry across the cell membrane. PMID- 2561785 TI - Desensitization of the acetylcholine receptor of frog end-plates measured in a Vaseline-gap voltage clamp. AB - 1. Desensitization of the nicotinic acetylcholine receptor of the frog end-plate was investigated in dissociated frog muscle fibres using the Vaseline-gap clamp method so that a wide range of well-defined agonist concentrations could be used without having to use alpha-bungarotoxin to reduce currents, and so that the intracellular medium could be controlled. 2. Acetylcholine (ACh) concentrations between 1 and 1000 microM were used, after inactivation of acetylcholinesterase. The intracellular calcium concentration was usually kept near zero by using 80 mM K2EGTA as the intracellular solution. 3. When using the low intracellular calcium solution, desensitization proceeded as a biphasic process with estimates of fast and slow time constants of about 8 and 80 s at 4 degrees C and 20 microM-ACh (the rates increased with concentration). In contrast, only one (fast) component of desensitization was detected when the intracellular calcium concentration was allowed to increase during ACh application. 4. Despite rapid application of ACh the time to peak response was 0.2 s (with 400 microM-ACh) to 2 s (with 1 microM ACh); this slow rise was shown to result from diffusion delays. Nevertheless the peak current with 200 microM-ACh corresponded to opening of most of the channels present, so there is probably not much desensitization in the millisecond time range. 5. Both fast and slow time constants for onset of desensitization showed only slight dependence on membrane potential when [Ca2+]i was buffered with 80 mM K2EGTA. 6. Increasing the intracellular cyclic AMP concentration directly, or indirectly with forskolin and IBMX, had no effect on the time course of desensitization. 7. Intracellular application of submicromolar concentrations of phorbol-12,13-dibutyrate (PDBu) and phorbol-12-myristate-13-acetate (PMA) yielded a small but reproducible reduction of the peak response to ACh. The time course of desensitization was, however, not modified by these substances. 8. The implications of these observations for the mechanism of desensitization, and their relationship to single-channel observations, are discussed. PMID- 2561786 TI - The role of inactivation in the effects of n-alkanols on the sodium current of cultured rat sensory neurones. AB - 1. The whole-cell patch-clamp technique has been used to investigate the actions of n-butanol, n-pentanol, n-hexanol and n-octanol on the sodium current of cells isolated from the dorsal root ganglia (DRGs) of neonatal rats and maintained in short-term tissue culture. 2. The influence of n-alkanols on the level of steady state inactivation of the sodium current was investigated by a standard two-pulse protocol. All alkanols increased the level of resting inactivation and this was manifested as a hyperpolarizing shift of the relationship between the steady state inactivation parameter (h infinity) and membrane potential. The mid-point of the h infinity curve was moved by up to -30 mV. 3. The relationship between the shift in the mid-point of the inactivation curve (delta Vh) and aqueous n alkanol concentration has been derived for each n-alkanol. These are complex in shape and do not appear consistent with a hypothesis that the increase in inactivation results from 1:1 binding of an alkanol molecule to a single site on the channel protein. 4. The aqueous concentrations used ranged from 70 mM-n butanol to 0.05 mM-n-octanol. However, equal fractional saturations of n-alkanols produced approximately equal shifts in the h infinity curve, particularly in the range 0.01-0.07 saturated. This implies a hydrophobic site of action, with a standard free energy per methylene group for adsorption to the site from the aqueous phase of ca -3.2 kJ/mol. 5. The increase in resting inactivation was not the sole means by which n-alkanols reduced the sodium current. The current was still reduced in cells pre-pulsed to sufficiently negative potentials to remove steady-state inactivation even in the presence of alkanols. The concentration required to reduce the current by 50% (ED50) has been interpolated for each n alkanol. From these data it was estimated that the standard free energy per methylene group for adsorption to the site of action was ca -3.1 kJ/mol, similar to that calculated for the effect on inactivation. The concentration dependence of this residual block indicated the involvement of more than one n-alkanol molecule. 6. The n-alkanols increase the level of inactivation of rat DRG cell sodium channels at potentials around the resting membrane potential and this effect contributes to their local anaesthetic action.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2561787 TI - Modulation of the delayed rectifier potassium current in frog cardiomyocytes by beta-adrenergic agonists and magnesium. AB - 1. The regulation of IK and ICa were studied in single cells isolated from bull frog atrium using the whole-cell configuration of the patch clamp and a perfused patch pipette. 2. IK was increased approximately 50-100% and ICa was increased approximately 6-10 times by 1 microM-isoprenaline, 5 microM-forskolin, or internal perfusion with 30 microM-cyclic AMP. The effects of cyclic AMP and isoprenaline were not additive. The shape of the concentration-response curves and the EC50 values for the effects of cyclic AMP on ICa and on IK were very similar (2.3 microM for IK and 1.7 microM for ICa). 3. Elevation of intracellular cyclic AMP had a similar effect on IK regardless of whether ICa was blocked with Cd2+ or not. Increasing ICa with dihydropyridine Ca2+ channel agonists had no effect on IK amplitude. 4. Isoprenaline or cyclic AMP caused an increase in the fully-activated IK and also shifted the activation curves to more negative potentials in most cells. The shift in the activation curve was reversible and was also observed when ICa was blocked with Cd2+. The rate of activation of IK was increased and the rate of deactivation of IK was slowed by isoprenaline. 5. After breaking the membrane patch and initiating whole-cell recording, IK ran down with time in about 50% of the cells examined when the intracellular solution contained 1 mM [Mg2+]. In contrast, when the solution contained 0.3 mM [Mg2+], rundown was almost never observed. Internal perfusion with increasing concentrations of [Mg2+] caused reversible decreases in the maximum amplitude of IK and shifted the IK activation curve slightly to more negative potentials, but had negligible effects upon the shape or the curvature of the fully activated current-voltage relationship. PMID- 2561788 TI - Modulation of excitatory amino acid receptors by group IIB metal cations in cultured mouse hippocampal neurones. AB - 1. Responses to the excitatory amino acids kainate, quisqualate, N-methyl-D aspartate (NMDA), L-glutamate and L-aspartate were recorded in mouse hippocampal neurones in cell culture, using the whole-cell configuration of the patch clamp technique. Agonists were applied rapidly from an array of flow pipes each of 250 microns diameter, positioned within 100 microns of the nerve cell body. 2. Responses to NMDA, L-aspartate and to low concentrations of L-glutamate, recorded with glycine in the extracellular fluid, were strongly antagonized by 50 microM zinc. Responses to kainate, quisqualate, and in glycine-free solution, responses to L-glutamate, were potentiated by 50 microM-zinc, but partially antagonized by 1 mM-zinc. On average, with 50 microM-zinc, responses to NMDA were reduced to 0.19 times control, while responses to kainate and quisqualate were increased to 1.09 and 1.14 times control. With 1 mM-zinc responses to kainate and quisqualate were reduced to 0.54 and 0.42 times control. 3. Cadmium had a similar, though less potent action, and at 50 microM antagonized responses to NMDA but potentiated responses to kainate and quisqualate. On average, with 50 microM cadmium, responses to NMDA were reduced to 0.39 times control, while responses to kainate and quisqualate were increased to 1.08 and 1.15 times control. With 1 mM cadmium responses to NMDA were reduced to 0.04 times control while responses to kainate and quisqualate were reduced to 0.79 and 0.60 times control. Mercury was neurotoxic and increased the leakage current; however, no reduction of the response to NMDA was produced by 5 microM-mercury. 4. The equilibrium dissociation constant (Kd) for zinc antagonism of responses to NMDA, estimated from fit of a single binding site adsorption isotherm, was 13 microM; cadmium was about 4 times less potent than zinc. These effects of zinc and cadmium were nearly voltage independent. In contrast the antagonism of responses to NMDA by 150 microM-magnesium was highly voltage dependent, such that the Kd for magnesium increased e-fold per 17.6 mV depolarization. 5. The potency of zinc as an NMDA antagonist did not vary with the concentration of NMDA, and was not greatly influenced by a 1000-fold variation in the concentration of the NMDA-modulator glycine. This suggests that zinc acts as a non-competitive antagonist, and does not directly interfere with the binding of NMDA to the agonist recognition site nor with the binding of glycine to an allosteric site on the NMDA receptor complex.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2561789 TI - The action of zinc on synaptic transmission and neuronal excitability in cultures of mouse hippocampus. AB - 1. The whole-cell configuration of the patch clamp method was used to record from hippocampal neurones in cell culture. Synaptic responses were evoked by loose patch stimulation of adjacent presynaptic neurones in low-density cultures. Agonists and antagonists were applied rapidly, using an array of flow pipes each of diameter 250 microns, positioned within 100 microns of the postsynaptic neurone. 2. Bath application of 50 microM-zinc produced prolonged periods of synaptic barrage and action potential discharge. Flow pipe application of 50 microM-zinc, in glycine-free solution with 1 mM-Mg2+, produced on average a 75% reduction of IPSP amplitude, but increased the average EPSP amplitude to 171% of control. However, after block of gamma-aminobutyric acid (GABA) receptors with bicuculline, zinc had no effect on EPSP amplitude, suggesting that potentiation recorded in control solutions reflects block of polysynaptic IPSPs. 3. Consistent with the block of IPSPs postsynaptic responses to flow pipe applications of GABA were blocked by zinc, with fast-on, fast-off kinetics. The equilibrium dissociation constant (Kd) for zinc block of GABA responses, estimated from fit of a single binding site adsorption isotherm, was 11 microM and sufficient to explain the degree of reduction of IPSPs by 50 microM-zinc. Zinc antagonism of responses to GABA was essentially independent of membrane potential over the range -60 to +60 mV. 4. With bicuculline methiodide and glycine added to a magnesium-free extracellular solution, to allow the study of synaptic responses mediated by N-methyl-D-aspartic acid (NMDA) receptors, zinc reduced the amplitude of EPSPs to 50% of control, and decreased the decay time constant of the EPSP, suggesting that zinc blocks synaptic activation of NMDA receptors. 5. Under conditions where synaptic transmission was completely blocked with postsynaptic receptor antagonists (1-3 mM-kynurenic acid and 10-20 microM-bicuculline methiodide) 50 microM-zinc decreased the amplitude of the spike after hyperpolarization (AHP), but did not produce large changes in action potential amplitude or half-width. Under these conditions 50 microM-zinc also decreased the current threshold required to trigger action potential discharge, and blocked accommodation so that repetitive firing replaced single action potential responses to prolonged current pulses. PMID- 2561790 TI - Characteristics of voltage-gated Ca2+ currents in ovine gonadotrophs. AB - 1. Voltage-clamp recordings were obtained from gonadotrophs of the ovine pars tuberalis in dissociated cell culture, utilizing the whole-cell recording mode of the patch-clamp technique. 2. The amplitudes of Ca2+ and Ba2+ currents were dependent on the extracellular concentration of divalent cation. 3. Ba2+ tail currents were observed on termination of depolarizing voltage steps. The extrapolated amplitudes of 'instantaneous' tail currents increased with membrane depolarization and showed saturation beyond +15 mV. 4. True inactivation of currents occurred in the presence of both external Ca2+ and Ba2+, judged from decrease in tail current amplitudes with progressive increases in duration of the activating voltage pulse. The inactivation process was fitted by a single exponential function at membrane potentials below -25 mV, while at more depolarized potentials the inactivation was better described by a double exponential function. The inactivation time constants decreased with positive shifts in membrane potential favouring a voltage-dependent inactivation. 5. The half-value of steady-state inactivation was observed at -40 mV using a two-pulse protocol. 6. Power spectral analysis of Ba2+ current noise from the steady-state portion of inward current showed a double Lorentzian fit of the power spectrum. 7. Two types of voltage-activated Ca2+ currents were identified based on their kinetics, voltage dependence, dependence on activation frequency, differential sensitivity to intracellular ATP and cyclic AMP, and to extracellular application of nifedipine. The channels with faster kinetics had a lower activation threshold (-50 mV) and the amplitude of the current was sensitive to clamping frequency. 8. From ensemble noise analysis of mean maximal inward current, single-channel amplitude of about 1 pA was estimated in 50 mM-Ba2+. PMID- 2561791 TI - Sodium-dependent membrane current induced by carbachol in single guinea-pig ventricular myocytes. AB - 1. In the presence of either barium (0.2 mM) or caesium (20 mM), carbachol (3-300 microM) depolarized isolated guinea-pig ventricular myocytes. Carbachol induced an inward current under voltage clamp at a holding potential equal to the resting potential (-75 mV). 2. Acetylcholine and oxotremorine also evoked an inward current but were less effective than carbachol. Atropine (0.3 microM) prevented the depolarization and inward current induced by carbachol and acetylcholine but not by oxotremorine. Moreover, oxotremorine, but not carbachol, induced an inward current in the absence of extracellular sodium. 3. Carbachol increased membrane chord conductance when it induced an inward current. These effects were recorded under experimental conditions that suppressed the voltage- and time-dependent sodium current (tetrodotoxin) and calcium current (cadmium), the inwardly rectifying potassium current, iK1 (caesium, barium and tetraethylammonium) and the current generated by the sodium-potassium pump (zero external potassium). 4. Under these same experimental conditions, the steady-state I-V relationship in the presence of carbachol was subtracted from that in its absence. The apparent reversal potential (Erev) was 25 mV with extracellular Na+ ([ Na+]o) at 143 mM and intracellular Na+ ([Na+]i) at 11 mM. Replacement of [Na+]o by N-methyl-D glucamine was associated with a shift of the apparent Erev to more negative voltages by approximately 61 mV per tenfold change of [Na+]o. 5. Isoprenaline induced an inward current in ventricular myocytes that depended upon sodium entry, required the accumulation of cyclic AMP and which was partially suppressed by acetylcholine (Egan, Noble, Noble, Powell, Twist & Yamaoka, 1988). In contrast to the current evoked by beta-adrenoceptor agonist, the current induced by muscarinic agonist was smaller and sustained. Moreover, the carbachol-induced current was not suppressed by prior addition of isoprenaline. 6. The findings are consistent with the mechanism that carbachol activates a plasma membrane ion channel that admits sodium and thereby increases intracellular sodium activity. The estimated increase of intracellular sodium activity from electrophysiological data agrees quantitatively with that obtained from measurements with sodium sensitive microelectrodes (Korth & Kuhlkamp, 1985). 7. The ability of carbachol to increase sodium influx may be the first step in a series of reactions that eventually alters sodium-calcium exchange and could account for catecholamine independent stimulation of developed force in mammalian ventricle. PMID- 2561792 TI - Single sodium channels from canine ventricular myocytes: voltage dependence and relative rates of activation and inactivation. AB - 1. Single sodium channel currents were recorded from canine ventricular myocytes in cell-attached patches. The relative rates of single-channel activation vs. inactivation as well as the voltage dependence of the rate of open-channel inactivation were studied. 2. Ensemble-averaged sodium currents showed relatively normal activation and inactivation kinetics, although the mid-point of the steady state inactivation (h infinity) curve was shifted by 20-30 mV in the hyperpolarizing direction. This shift was due to the bath solution, which contained isotonic KCl to depolarize the cell to 0 mV. 3. Steady-state activation showed less of a voltage shift. The threshold for eliciting channel opening was around -70 mV and the mid-point of activation occurred near -50 mV. 4. The decline of the ensemble-averaged sodium current during a maintained depolarization was fitted by a single exponential function characterizing the apparent time constant of inactivation (tau h). The apparent rate of inactivation was voltage dependent, with tau h decreasing e-fold for a 15.4 mV depolarization. 5. The relative contributions of the rates of single-channel activation and inactivation in determining the time course of current decay (tau h) were examined using the approach of Aldrich, Corey & Stevens (1983). Mean channel open time (tau o) showed significant voltage dependence, increasing from 0.5 ms at -70 mV to around 0.8 ms at -40 mV. At -70 mV tau h was much greater than tau o, while at -40 mV the two time constants were similar. 6. The degree to which the kinetics of single-channel activation contribute to tau h was studied using the first latency distribution. The first latency function was fitted by two exponentials. The slow component was voltage dependent, decreasing from 19 ms at 70 mV to 0.5 ms at -40 mV. The fast component (0.1-0.5 ms) was not well resolved. 7. Comparing the first latency distribution with the time course of the ensemble averaged sodium current at -40 mV showed that activation is nearly complete by the time of peak inward sodium current. However, at -70 mV, activation overlaps significantly with the apparent time course of inactivation of the ensemble averaged current. 8. Using the methods of Aldrich et al. (1983) we also measured the apparent rate of open-channel closing (a) and open-channel inactivation (b). Both rates were voltage dependent, with a showing an e-fold decrease for an 11 mV depolarization and b showing an e-fold increase for a 30 mV depolarization.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2561793 TI - Electrophysiological properties of rat CA1 pyramidal neurones in vitro modified by changes in extracellular bicarbonate. AB - 1. Intracellular recordings were made from the somata of CA1b hippocampal pyramidal neurones in vitro and the concentration of bicarbonate ion ([HCO3-]o) in the artificial cerebrospinal fluid (ACSF) was varied by substitution for Cl-. 2. Reducing [HCO3-]o from 26 mM (standard ACSF) to 8.6 mM or raising it to 72 mM had only minor effects on resting membrane potential but input resistance was reduced and increased, respectively. Threshold for Na(+)-dependent action potential generation was raised during the perfusion with low-HCO3- ACSF and lowered during perfusion with high-HCO3- ACSF. In tetrodotoxin-poisoned neurones where presumed Ca2(+)-dependent potentials could be elicited in standard ACSF, perfusion with high-HCO3- ACSF lowered the activation threshold. Where no Ca2(+) dependent spikes could be elicited in standard ACSF, perfusion with high-HCO3- ACSF caused their appearance. Ca2(+)-dependent spikes could not be evoked during perfusion with low-HCO3- ACSF. 3. Depolarizing current pulses evoked two basic patterns of action potential discharge under standard [HCO3-]o conditions. Conversion between the two types was possible by varying [HCO3-]o. In those neurones which fired a train of fast Na(+)-dependent spikes in standard ACSF, perfusion with high-HCO3- ACSF usually led to the development of burst discharges. A smaller number of neurones responded to depolarizing current with an initial burst of three to six action potentials; the bursts were attenuated in low-HCO3- ACSF and replaced by a repetitive spike discharge. Frequency accommodation of spike discharge in response to depolarizing current pulses and the after-hyperpolarization following a current-evoked discharge were usually both attenuated in low-HCO3- ACSF and enhanced in high-HCO3- ACSF. 4. Orthodromically evoked excitatory postsynaptic potentials (EPSPs) and early and late inhibitory postsynaptic potentials (IPSPs) were reduced in amplitude during perfusion with low-HCO3- ACSF. In high-HCO3- ACSF, EPSP amplitude and duration increased, the latter reflecting a positive shift in the reversal potential of the early IPSP consequent upon reduced [Cl-] in high-HCO3- ACSF. The late IPSP was, however, unaffected. 5. Responses to ionophoretically applied excitatory amino acids were enhanced in high-HCO3- ACSF and depressed in low-HCO3- ACSF. 6. Perfusion with high-HCO3- ACSF was associated with the development of epileptiform activity. Spontaneous or synaptically evoked bursts of action potentials were indistinguishable and could be blocked by N-methyl-D-aspartate antagonists.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2561795 TI - [Interaction of antibodies with cardiac neurotransmitter receptors]. AB - This paper demonstrates that antibodies against neurotransmitter receptors of the cardiac membrane are able to alter the physiology, pharmacology and biochemistry of the target organ. Sera from chagasic patients contain an antibody which binds to beta adrenoceptors of myocardium and modulates their activity. Chagasic IgG simulates a partial beta agonist by increasing contractility and diminishing reactivity to exogenous norepinephrine. Moreover, chagasic IgG inhibits the binding of the specific radioligand to purified cardiac membranes. The interaction of chagasic IgG with beta-adrenoceptors triggers signal transduction resulting in the stimulation of adenylate cyclase with an increased production of cAMP. Stimulation of Ca(++)-ATPase and inhibition of Na+ + K(+)-ATPase are also observed. These enzyme dysfunctions induce modifications of cellular thermodynamic equilibrium that trigger both morphological and functional alterations of the myocardium. Other immune sera can also trigger pharmacologic effects on isolated atria. Immune IgG directed against specific alloantigens and IgG from mice with autoimmune myocarditis are able to recognize the beta adrenergic receptor-coupled adenylate cyclase system and alter the function of target organs. The detection of antibodies against neurotransmitter receptors could be a useful marker during the early stages of development of autoimmune diseases. PMID- 2561796 TI - The effect of pyridoxine on the depressant action of ethanol. PMID- 2561794 TI - VIP-helodermin receptors in the murine virus-induced T lymphoma cell line BL/VL3 recover less rapidly than beta-adrenoceptors after down regulation. AB - The time course of recovery of beta-adrenergic and VIP/helodermin receptors after homologous and heterologous down regulation was studied in the murine lymphoma cell line BL/VL3, a neoplastic equivalent of immature T cells. The heterologous part of isoproterenol down regulation was rapidly reversed, even in the presence of cycloheximide, suggesting that down regulation was linked to ligand-receptor interaction and/or cyclic AMP increase. Homologous down regulations of beta adrenoceptors and VIP/helodermin receptors were less rapidly reversible and depended on protein synthesis as they were inhibited by cycloheximide: beta adrenoceptors recovered faster than VIP/helodermin receptors. PMID- 2561798 TI - [Serologic study of arbovirus in 2 localities of the Juventud island]. AB - A serologic study is made in two population groups in the Isle of Youth. A total 268 blood samples in blotting paper are subjected to the hemagglutination inhibition technique, using the Eastern equine encephalomyelitis, Western equine encephalomyelitis, Saint Louis encephalitis, and dengue 2 viruses; 16% positivity to flavivirus was found. A second serum sample was taken in people positive by the hemagglutination inhibition technique in order to carry out the techniques of complement fixation and plate reduction neutralization. Nine cases showed complement-fixating antibodies, which is indicative of recent infection and in 29 cases neutralizing antibodies to SLE virus were found. PMID- 2561797 TI - [Prevalence of HBSAG in blood donors and patients at the central hospital of Maputo, Mozambique, 1985]. AB - A study is made of 681 donors who came to the Blood Bank of Maputo Central Hospital in Mozambique during the months of November, 1984, to January, 1985. By means of the technique of counterimmunoelectrophoresis it was determined that 18.6% of donors were positive for Ag HBs. Likewise, 90 patients were analyzed; these patients were hospitalized in the Gastroenterology Service of the hospital with different clinical diagnoses: primary liver carcinoma (14), acute hepatitis (24), cirrhosis (10), chronic hepatitis (7), and others (35). 41.6% of the patients hospitalized with hepatitis were type B. There was a significant relationship between Ag HBs carriers and patients with primary liver carcinoma. PMID- 2561799 TI - [Comparison of latex agglutination technics and polyacrylamide gel electrophoresis for the rotavirus diagnosis. Habana City, 1986]. AB - Two methods for the diagnosis of rotavirus are compared with 120 stool samples from children under two years of age who had been hospitalized in Centro Habana Children's Hospital with a clinical diagnosis of acute diarrheal disease and 30 samples of healthy children of the same age group who served as controls. The methods used were latex agglutination (Rotalex, commercially-available kit from ORION DIAGNOSTIC ESPOO, Finland) and polyacrylamide gel electrophoresis. Electronic microscopy was used to define those cases whose results did not coincide with the previous techniques. Comparison was made based on the sensitivity and specificity, cost and easiness of performance. Rotalex was less specific but more sensitive than electrophoresis in polyacrylamide gel. The latter is less expensive, since reactants are available at a lower price in dollars and have no expiration date, is very useful in epidemiologic and outbreak studies, since it detects subgroups of circulating rotavirus and it is a painstaking technique (1-20 samples in 10 hours). Rotalex is simple, rapid (1-20 samples in 30 minutes) and allows individual determinations; therefore, we consider its application in hospitals is feasible. PMID- 2561800 TI - [Rotavirus infection in children with acute diarrhea in Habana City]. AB - This paper shows the incidence of rotavirus and other pathogenic agents in 256 children under three years of age with a diagnosis of acute diarrhea. This study included the months of December 1983 through May 1984. Rotavirus was found in 27.7% of patients, followed by enteropathogenic E. coli (17%). No positive cases were found in the control group. The highest incidence of rotavirus infection coincided with the coldest months where the lowest volumes of rains were reported. PMID- 2561802 TI - Experimental models for studying the microbial ecology in the intestinal tract. PMID- 2561801 TI - [The incidence of enterotoxigenic Escherichia coli, rotavirus and Clostridium perfringens from cases of diarrhea in children, in the region of Campinas, SP, Brazil]. AB - A survey for the detection of enterotoxigenic Escherichia coli (ETEC), rotavirus and enterotoxigenic Clostridium perfringens in diarrheic stools of children up to 2 years old was carried out in the region of Campinas, SP, Brazil. Twenty-seven (20.45%) faecal specimens were positive for ETEC. From these samples 41 strains of ETEC were isolated from which 40 produced only thermolabile (LT) enterotoxin, as detected by a modified radial immune haemolysis test. Among the 183 faecal specimens examined for the detection of rotavirus, 29 (15.84%) were positive when examined by polyacrilamide gel electrophoresis (PAGE) and immunoenzymatic assay (EIA) being 15 (51.7%), derived from stools collected from winter months. All strains of rotavirus belonged to group A and through the PAGE technique, it was observed that the most frequent (9 strains) electrophoretype, according to the adopted classification, was Ib, IIc, IIIb, IVa. Only 113 fecal specimens were examined for the presence of enterotoxigenic C. perfringens. For the detection of enterotoxin in culture supernatants the reverse passive haemagglutination and intravenous inoculation of mice were used. Twelve (10.61%) enterotoxigenic C. perfringens strains were found. Taking into consideration these findings the authors call the attention of the relative value of conventional coprocultures for diagnostic purposes, pointing out the important of establishing simplified methods which would render easier, the detection and identification of the groups of enteropathogenic agents studied in this research. PMID- 2561803 TI - Signs, symptoms, and significance of paraneoplastic neurological syndromes. AB - Neurological lesions attributed to the remote effect of cancer on the nervous system are protean, involving any part of the central and/or peripheral nervous system including the muscles. These paraneoplastic neurological syndromes are relatively uncommon in cancer patients, their relative frequency is variable, and their pathogenesis remains largely unknown. All forms of PNNS have been seen both in patients with and without cancer, and, unlike the paraneoplastic endocrinopathies, tumor removal is not necessarily followed by the remission of PNNS. For some PNNS, specific pathogenic mechanisms have been established or are highly probable. Thus, certain metabolic encephalopathies or the carcinoid myopathy are due to production of hormones or hormonelike substances by the tumor. The discovery of the most "typical" forms of PNNS justifies a careful search for an underlying neoplasia. PMID- 2561804 TI - Regulation of the polyphosphoinositide-specific phosphodiesterase in B lymphocytes. PMID- 2561806 TI - Alternative self or nonself recognition of an antigen expressed in a rare cell type in transgenic mice: implications for self-tolerance and autoimmunity. PMID- 2561805 TI - Signal transduction by the antigen receptor of B lymphocytes. PMID- 2561807 TI - [Application of granulate hydroxylapatite for mandibular augmentation]. AB - The already known surgical interventions for absolute induction of extensive mandibular atrophy, cannot be suggested as routine standardized methods. Use of Hydroxylapatite-ceramic for restoration of the atrophic alveolar process, shows several advantages; among those, simplicity of the operation use of local anesthesia, as well as very low percentage of unsuccessful interventions are taken into consideration. The clinical results of 102 cases are shown and discussed. Through the addition of H.A. ceramic in the field of preprosthetic surgery, there is a broadening of the spectrum of the therapeutic possibilities of patients with prosthetic problems to be expected. PMID- 2561808 TI - Odontodysplasia--a unilateral dental malformation. PMID- 2561809 TI - [Usefulness of the passive hemagglutination micromethod in the diagnosis of whooping cough]. AB - Studies on modification of diagnostic test for pertussis have been continued, they were practically restricted to an application of passive haemagglutination micromethod. Six hundred and twenty eight sera from children suspected to be infected with Bordetella and 38 sera of control children suffering from non infectious diseases were tested. Despite of the fact that higher titers were obtained with method using red blood cells preserved in Alsevier solution, the passive haemagglutination micromethod may be used in field studies especially during pertussis epidemics what was confirmed by statistical analysis. PMID- 2561810 TI - [Detection of toxins of Clostridium perfringens type A in infected animals by using the ELISA test]. AB - The aim of this study was to evaluate the usefulness of ELISA in toxin detection in guinea pigs experimentally infected with toxinogenic strain of Clostridium perfringens type A. The toxin was detected in blood serum and muscles from 12 hours after infection. The results obtained indicate the advantage of ELISA over to date methods used as immunofluorescence or microscopic examination of muscle exudate or sections. ELISA due to its high sensitivity rapidity and specificity allows to detect toxin in guinea pigs before clinical symptoms of gas gangrene are developed. PMID- 2561811 TI - [Effect of itraconazole on the process of mycelial transformation of Candida albicans cells in human serum]. AB - Results of in vitro studies on the influence of itraconazole on mycelial transformation of 88 Candida albicans strains are presented. The drug influenced a number of cells capable to produce filaments. The number of those cells diminished gradually together with the increase of the drug concentration. The range of TI100 values was between 0.02 and 18.0 micrograms ml of medium. Mycelial transformation phenomenon can be applied for preliminary and rapid evaluation of Candida albicans strains sensitivity to itraconazole. PMID- 2561812 TI - [Hormonal action on the physiology of osteoblasts]. AB - In this review the recent knowledge of the hormonal control of osteoblastic cells in presented. I refer to the action of parathormone, vitamin D3, glucocorticoids, calcitonin and other hormones stating the very recent experimental works, and I make speculations about the possible mechanism of bone resorption as a result of hormonal action. So far, osteoblasts are more and more implicated in this mechanism during pathologic resorption or physiological bone turnover. PMID- 2561813 TI - [Prospect of avian encephalomyelitis antibodies in breeding farms in Chile]. AB - Ten different layer and broiler breeding farms, located in the central part of the country were sampled to detect antibodies against the Avian Encephalomyelitis Virus (AEV). Fifty embrionated eggs, per house, were examined using the Embryo Susceptibility Test. Samples of eggs from 21 houses of broiler breeder hens and 9 of breeder layers were collected from all the poultry farms studied. The antigen used in the Embryo Susceptibility Test was prepared from the van Roekel strain for AEV. The results indicated a high degree of specific immunity against the AEV in 86.7% of the sections checked. This immunity could be due to the presence of the causal of AEV in natural form, since the birds in these farms had not been vaccinated against this ornithopathy. This is the first study carried out in Chile which detected the presence of antibodies against AEV in layer and broiler breeding farms, leading to a massive vaccination program against the disease in the country. PMID- 2561815 TI - Plasma and red blood cell magnesium concentrations in Zucker rats: influence of a high fibre diet. AB - Because of the possible influence of obesity and insulin levels on magnesium metabolism, we have examined plasma and red blood cell (RBC) Mg in Zucker rats. There were reduced plasma and RBC Mg levels in Zucker rats compared with Wistar rats, and phenotype differences in Zucker rats. In obese rats, a high fibre diet reduced body weight, hyperinsulinaemia and hyperlipaemia and returned the plasma Mg concentration toward normal. However RBC levels were significantly lower than in lean rats of the same strain. The significance of the altered Mg concentration in obese Zucker rats may have metabolic implications and needs further study. PMID- 2561816 TI - Electron microscopic detection of human papillomavirus particles in oral proliferative lesions. AB - Human papilloma virus (HPV) has been demonstrated in a series of benign proliferative lesions of skin and mucosae. To prove the distribution of HPV in the oral proliferative lesions at the ultrastructural level, we performed electron microscopic analysis of 10 specimens taken from 5 patients through large excisional biopsy. All of them were diagnosed pathologically as fibropapilloma. In each patient, specimens were taken from both clinically evident proliferative lesions and clinically normal surrounding mucosa. Obtained specimens were fixed in a glutaraldehyde solution and processed for routine ultrathin sectioning. Before electron microscopic observation, the tissue sections on copper grids were subjected to amylase digestion of glycogen granules. Spherical viral particles of 40-55 nm in diameter were detected the non-keratinized epithelial cells in all specimens examined. Of particular interest were the large amounts of viral particles found in the cytoplasmic matrix and nuclei (especially on their chromatin masses) of the cells in intermediate and surface layers, which did not form a crystal array. All the membranous cell organelles of epithelial cells were, however, devoid of viral particles. Some viral particles were distributed in the extracellular spaces of an intermediate layer. Viral particles were hardly observed in the cells of a basal/suprabasal and prickle cell layers. There were no significant differences in the HPV distribution between the cells derived from the proliferative lesion and those derived from the surrounding normal mucosa. PMID- 2561814 TI - Malignant B cell immunoblastic lymphoma expressing CD30 antigen in the terminal phase of a multiple myeloma. AB - An acute terminal phase of a Bence Jones kappa plasma cell myeloma developed 22 months after chemotherapy is presented. The patient's symptoms were fever, cytopenias, adenomegalies and hepatomegaly. A lymph node biopsy showed an immunoblastic polymorphic kappa B cell lymphoma, expressing CD30 antigen. After new polychemotherapy the patient died because of bleeding and infection. Autopsy revealed a decrease in the tumour with defective immunophenotype. The few reported cases have been reviewed with emphasis on clinical aspects, prognosis and morphology, The significance of CD30 positivity (activation marker) in a high grade lymphoma is discussed. PMID- 2561817 TI - [Compression syndromes in peripheral nerves]. PMID- 2561818 TI - Arthritic inflammation in rats as a model of chronic pain: role of opioid systems. PMID- 2561819 TI - Disruption of schedule-controlled behavior during abstinence from phencyclidine and tetrahydrocannabinol. PMID- 2561821 TI - Plasma delta-9-THC levels as a predictive measure of marijuana use by women. PMID- 2561820 TI - Behavioral and EEG studies of acute cocaine administration: comparisons with morphine, amphetamine, pentobarbital, nicotine, ethanol and marijuana. PMID- 2561822 TI - Opioid receptors and ligands. PMID- 2561823 TI - The role of opioids in analgesia and gastrointestinal function. PMID- 2561824 TI - Opioid delta receptor involvement in behavioral and neural plasticity. PMID- 2561825 TI - Thermoregulation and the opioid system. PMID- 2561826 TI - Site-directed affinity ligands as tools to study the phenomenology and mechanisms of morphine-induced upregulation of opioid receptors. PMID- 2561827 TI - Mu antagonist and kappa agonist properties of b-funaltrexamine (b-FNA): long lasting spinal antinociception. PMID- 2561828 TI - Cocaine binding sites related to drug self-administration. PMID- 2561829 TI - Mechanisms of phencyclidine (PCP)-n-methyl-d-aspartate (NMDA) receptor interaction: implications for drug abuse research. PMID- 2561831 TI - Behavioral pharmacology of PCP, NMDA and sigma receptors. PMID- 2561830 TI - Characterization of the actions of phencyclidine on midbrain dopamine neurons. PMID- 2561832 TI - Cannabinoid modulation of cyclic AMP accumulation in synaptosomes. PMID- 2561833 TI - [3H]AHN 086 acylates peripheral benzodiazepine receptors in the rat pineal gland. PMID- 2561834 TI - A new analysis of whole-body calorimetry data. PMID- 2561835 TI - Irreversible affinity ligands for mu opioid receptors. PMID- 2561836 TI - Delta opioid receptor selective alkaloid agonists and antagonists. PMID- 2561837 TI - Evidence for rapid development and loss of opioid tolerance to fentanyl in the rat. PMID- 2561838 TI - Effects of delta-9-THC on repeated acquisition and performance of response chains in humans. PMID- 2561839 TI - Urinary elimination half-life of delta-1-tetrahydrocannabinol-7-oic acid in heavy marijuana users after smoking. AB - The urinary excretion of delta 1-tetrahydrocannabinol-7-oic acid (delta 1-THC-7 oic acid), the major urinary metabolite of delta 1-THC, and the total amount of THC metabolites was studied in heavy marijuana users after smoking using high performance liquid chromatography and the EMT-d.a.u. cannabinoid assay. An average elimination half-life (+/- SD) of 3.0 +/- 2.3 days was obtained for delta 1-THC-7-oic acid. The average ratio (+/- SD) of "EMIT readings"/delta 1-THC-7-oic acid concentrations was 1.23 +/- 1.03. PMID- 2561840 TI - Effects of acute and chronic administration of (+)SKF 10,047 on body temperature in the rat. PMID- 2561841 TI - Potential neurotoxic effects of self-administered cocaine on dopamine receptors. PMID- 2561842 TI - Energy substrate metabolism in testis of rats treated with delta-9 tetrahydrocannabinol (THC) and cocaine (COC). PMID- 2561843 TI - Structural and conformational aspects of the binding of aryl-alkyl amines to the phencyclidine binding site. PMID- 2561844 TI - Cocaine increases benzodiazepine receptors labeled in the mouse brain in vivo with [3H]Ro 15-1788. PMID- 2561845 TI - Kappa antagonist effects of buprenorphine in the rat drug-discrimination procedure. PMID- 2561846 TI - Fluorescent probes for peripheral-type benzodiazepine receptor visualization and localization. PMID- 2561847 TI - Effects of mixed-action opioids on food-maintained behavior of morphine pretreated and morphine-tolerant rats. PMID- 2561848 TI - The effects of phencyclidine on the pituitary-adrenal axis are centrally mediated in the rat. PMID- 2561849 TI - Effects of naloxone and Mr 2266 on thermonociceptive reactions in diabetic mice. PMID- 2561850 TI - Characterization of NMDA-coupled and dopamine reuptake carrier coupled [3H]-TCP binding sites in guinea pig brain. PMID- 2561851 TI - Excitatory amino acid antagonists as well as GABA agonists cause barbiturate-like anesthesia in rats. PMID- 2561852 TI - Discrimination of the benzodiazepine receptor antagonist Ro 15-1788 using the conditioned taste aversion procedure. PMID- 2561853 TI - Cation requirement for GTP regulation of [125I]b-endorphin binding to Mu and sigma opioid receptors. PMID- 2561854 TI - Autoradiographic localization of 3H-dextromethorphan binding sites differs from NMDA. PMID- 2561855 TI - Changes in prodynorphin peptide content following treatment with morphine or amphetamine: possible role in mechanisms of action of drug of abuse. PMID- 2561856 TI - Modulation of synaptosomal free intracellular calcium in naive and morphine tolerant mice: correlation of calcium modulation in vitro and in vivo to tolerance development. PMID- 2561857 TI - In vivo binding of (+)-cis methylfentanyl at the opiate receptor complex and behavioral correlates: evidence for a novel mechanism of action. PMID- 2561858 TI - 1989 Annual Report, evaluation of new compounds for opioid activity. PMID- 2561859 TI - [HIV-positive newborns via maternal-fetal transmission. Early ophthalmic lesions]. AB - Since January 1986, the authors have examined twenty infant HIV positives. In two cases specially, some serious and precocious ophthalmic lesions have been found : an ophthalmic zona appeared at the age of three months, a cytomegalovirus retinitis diagnosed at the age of six months. From this series, the particularities of the HIV infection by maternal-fetal transmission are discussed. PMID- 2561860 TI - [Cytomegalovirus retinitis in acute leukemia]. AB - The authors reported a Cytomegalovirus retinitis in an acute leukemia. It appears differently in each eye: localised seats of retinitis in one eye, and central venous occlusion associated with central artery occlusion in the other eye. The clinical polymorphism of herpes group virus is discussed. PMID- 2561861 TI - [Presence of lysozyme and alpha 1-antichymotrypsin in tumors of the fibrous histiocytoma type. Immunohistochemical evaluation of 52 cases]. AB - In 52 cases of fibrohistiocytic sarcoma activity of alfa-1-antichymotripsin (ACT) in tumour cells was found in 47 cases and lysozyme in 16 cases. ACT was found in all types of tumour except angiomatous. Lysozyme was positive only in pleomorphic type, all these cases have also activity of ACT. Investigated enzymes are useful in diagnosis with correlation to others markers used in diagnostic procedure of soft tissue tumours. PMID- 2561862 TI - Vascular changes and inflammatory infiltrations in primary lung cancers. AB - Twenty one surgical specimens of lung cancer were studied. Microangiography showed that the vascularization of the tumours did not depend either on the size or histology of carcinoma. Abundant vascularity of tumours was associated with the degree development of connective tissue--vascular stroma. Poor vascularity was linked with neoplastic blockade of vessels and the presence of necrosis. Besides the cancer tumour the vascular changes existed in all parts of the lung, but they were expressed most profusely in the surroundings of neoplastic infiltrate. The range of histological vascular changes was as follows: elastosis, inflammation, lumen obliteration (proliferation of intima), wall thickening (arterialization of veins), oedema, hyalinization, plasmorrhagia, fibrosis (scarring), presence of acid GAG, thrombosis. The most striking vascular changes were connected with hilar and intralobar localization of the tumour, the mildest with subpleural position. The intensity of vascular changes depended on the size of tumour and existence of necrosis. The extent of necrosis and the histological type of neoplasm exerted no essential effects in this respect. Inflammatory cell infiltration appeared abundantly in the direct vicinity of the tumour in adenocarcinoma, with hilar localization and displayed proportional dependence on the size of tumour and the extent of necrosis, but the tumours, whose diameter exceeded 6 cm, failed to have any effect on the intensity of the inflammatory cell reactions. PMID- 2561863 TI - [Incidence of markers of HBV and HAV infections in workers of the departments and clinics of anesthesiology and intensive care at the medical academy in Wroclaw]. AB - The frequency of markers of infection by hepatitis virus A and B was studied among the workers (physicians, nurses, auxiliary personnel) of the Chair and Department of Anaesthesiology and Intensive Therapy, Medical Academy in Wroclaw. Serological tests were done by the immunoenzymatic method using kits of Abbott Diagnostics Division. IgG anti-HAV antibodies were found in 62.8% of the tested subjects. HBV markers were present in 44.3% of the subjects, that is several times more frequently than in the general population. HBsAg carriers accounted for 4.3% of the whole group, while in 40% of the group anti-HBV antibodies were present in various combinations suggesting an immunity against HBV infection. PMID- 2561864 TI - Physiological identification and property of afferents from the masseter muscle to the caudal part of the trigeminal nucleus in rat. AB - The present study was to identify the physiological properties of the afferent fibers from the masseter muscle to the caudal part of the spinal trigeminal nucleus of rat using a microelectrode technique. When electrical stimulation was applied to the masseter muscle, the evoked potentials were recorded with a shorter latency (the S-response) or a longer latency (the L-response). The threshold current intensity of the S-response was lower than that of the L response. There were statistical significant differences between the S-response and the L-response in latency and in threshold current intensity. Both the S- and L-responses could follow by stimulation of low frequencies (10 to 30 Hz), indicating that the evoked responses were the component of the secondary neuron activities. On the anatomical and physiological assumption, the conduction velocities of the S- and the L-responses were calculated and they were in the range of that of A-delta fibers. We also observed that both the S- and the L responses could not follow high frequency stimulation of the masseter muscle. This effect may reflect the phenomenon of fatigue in the finely myelinated fibers or polymodal nociceptors. Thus, the present study suggested the involvement of the afferents from the masseter muscle to the spinal trigeminal nucleus in the transmission and the relay of the masticatory muscle pain. PMID- 2561865 TI - Characteristics of responses in the trigeminal subnucleus caudalis and adjacent reticular formation evoked by the stimulation of the masseter muscle. AB - Our previous study reported that the afferents to the spinal trigeminal nucleus from the masseter muscle were predominantly of A-delta range fibers having fast conducting-low threshold and slow conducting-high threshold. In the present study, further experiment was undertaken to obtain more definite information on the central transmission of muscle pain. When the electrical stimulation was delivered to the masseter muscle of rat, the evoked potentials with a shorter latency (the S-response) or longer latency (the L-response) were recorded from the caudal part of the spinal trigeminal nucleus. In both the S- and the L responses, the relationship between the amplitudes of the responses and the stimulus intensity almost followed the power function with the exponent of about 3.4 and 4.0, respectively. The stimulus intensity range of the S-response, in which the power function was formed, was wider than that of the L-response. The most of the S-response were recorded from the trigeminal subnucleus caudalis and adjacent reticular formation, whereas the recording sites of the L-response were located mainly in the trigeminal subnucleus caudalis. These results suggest that the trigeminal subnucleus caudalis and adjacent reticular formation are an important portion of the afferent projection areas and of the modulator system of pain sensation from the masseter muscle. PMID- 2561866 TI - Novel method for the preparation of sterile collagen-agarose-plates for isolating collagenolytic bacteria. AB - An assay method for bacterial collagenase using sterile collagen-agarose plates was developed to isolate collagenolytic bacteria. In order to avoid heat denaturation of collagen, the plates were made at 37 degrees C mixing microwave sterilized collagen with low melting point-agarose. Sensitivities to various proteases were tested on the plates; it was shown that whereas the collagen in these plates was digested only by the collagenases, the collagen could also be hydrolyzed by other proteolytic enzymes if it had been either sterilized by ethylene oxide or solidified with conventional agar at a higher temperature. Colonies of Clostridium histolyticum cells grown on the plates had clear zones around them and their collagenase activities could be readily detected. PMID- 2561867 TI - [Tumor necrosis factor]. PMID- 2561868 TI - New developments on the generation of mutations in Escherichia coli lysogens. AB - Through the lytic process, P1 is a bacteriophage or virion capable at very low frequency of carrying out generalized transduction between strains of Escherichia coli. When P1 is not involved in lytic functions, it exists as a prophage in the form of circular DNA molecule, persisting in an extra-chromosomal or plasmid state, and not integrating into the host chromosome. E. coli carrying such plasmids have been referred to as lysogens. Earlier research dealt with P1 plasmids carrying drug-resistant factors. Up to the present study, P1 plasmids of any type were not known to have any mutagenic effect on the E. coli chromosome (genome), nor was it known that generalized transduction can be associated with mutagenicity. From P1 plasmids carrying a chloramphenicol resistance-factor, mutant plasmids of a particular type were isolated in the present study. P1 plasmids of this type carried a mutant factor which greatly impaired the capacity of phage derived from such plasmids, upon the completion of the lytic cycle, to lysogenize recA E. coli through phage promoted recombination. The plasmid of this mutant type is referred to as P1CMrec, and lysogens carrying such are referred to as P1CMrec lysogens. This paper describes the history of these P1CMrec lysogens and the genetic mutability within the E. coli chromosome of such P1CMrec lysogens, and the relationship of such genetic mutability (instability) to the incorporation of virion-DNA, following the absorption of P1 phage by these lysogens. As illustrated in the paper, a mutagenic effect was generated within the E. coli chromosome of P1CMrec lysogens by means of the P1CMrec plasmid. Furthermore, this mutagenic effect was found to be greatly, non-locally, and uniformly enhanced as a consequence of P1 virion incorporation by, or likely generalized transduction of, such lysogens. More specifically, the plasmid of this mutant type (P1CMrec) is responsible for the creation of a wide range of genetic mutabilities (instabilities) of differing degree within the E. coli genome (not carrying recA), some mutabilities being very high upon extended incubation. The P1CMrec plasmid was also involved in the creation of new mutant genes within the E. coli genome (not carrying recA), some mutabilities being very high upon extended incubation. The P1CMrec plasmid was also involved in the creation of new mutant genes within the E. coli chromosome, some of which manifested high mutability.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2561869 TI - NIMH seeks research on AIDS and mental health. PMID- 2561870 TI - Human papillomaviruses and cervical carcinoma. AB - The question of whether human papillomavirus (HPV) infection causes, or at least contributes to, the development of cervical carcinoma has been a topic of much scrutiny and controversy over the past few years. The sexual mode of transmission of genital HPVs and detection of viral DNA in cervical tumors suggest a causal role at some stage during malignant progression. To some extent it seems that the epidemiological data accumulated so far have served to confuse as much as to clarify the issue, and the distribution of genital HPVs in the general population appears to be more complex than originally anticipated. However, experimental systems that have been used to study HPVs, in particular recent studies utilizing human genital epithelial cells, have provided strong evidence that certain HPV types can cause premalignant changes in these cells. The development of cervical cancer is a multistage process, and HPV infection alone is clearly not sufficient for full malignant transformation. Nevertheless, identification and control of HPV infection may be of critical importance in the diagnosis and treatment of this disease. PMID- 2561871 TI - The polymerase chain reaction and cancer diagnosis. AB - The introduction of polymerase chain reaction (PCR) technology, and its continual refinement, have had an enormous impact on many areas of molecular biology, including cancer research. This review examines several types of PCR assays that have been designed to investigate the molecular basis of cancer, in particular, those assays that have potential diagnostic applications. PMID- 2561873 TI - Metaiodobenzylguanidine for cardiac and tumor imaging. PMID- 2561872 TI - Growth factor signaling pathways: phosphoinositide metabolism and phosphorylation of phospholipase C. AB - Recent demonstrations of growth factor-stimulated increases in cellular phosphoinositide metabolism suggest that regulatory enzymes of this important signaling pathway may be substrates for growth factor receptor tyrosine kinases. Studies employing phosphotyrosine antibodies, specific phospholipase C antibodies, and purified phospholipase C proteins support the conclusion that the 145-kD phospholipase C-gamma 1 isoenzyme is rapidly and selectively phosphorylated by the activated epidermal growth factor and platelet-derived growth factor receptors. The selective interaction of these receptors with phospholipase C-gamma 1 suggests a novel, direct mechanism for agonist stimulation of phosphoinositide metabolism. PMID- 2561874 TI - DOT-enzyme linked immunosorbent assay for detection of Clostridium perfringens type A enterotoxin. AB - A procedure, which we have termed DOT-ELISA, to detect Clostridium perfringens type A enterotoxin on nitrocellulose paper is described. Seventy eight preparations from 39 cultures of C. perfringens type A were tested simultaneously by this and by Plate-ELISA methods. The results were comparable. DOT-ELISA detected as little as 0.02 micrograms of purified enterotoxin and 0.13 micrograms of enterotoxin in cell-free culture supernatant. As little as 0.02 micrograms purified enterotoxin mixed with human faeces could be detected specifically. The method is simple and does not require an ELISA reader. PMID- 2561875 TI - Multisite phosphorylation of microtubule-associated protein 2 (MAP-2) in rat brain: peptide mapping distinguishes between cyclic AMP-, calcium/calmodulin-, and calcium/phospholipid-regulated phosphorylation mechanisms. AB - Microtubule-associated protein 2 (MAP-2), a cytoskeletal protein of 280 kilodalton that is highly enriched in dendrites and neuronal perikarya, is subject to both cyclic AMP-, calcium/calmodulin-, and calcium/phospholipid regulated phosphorylation when incubated with [gamma-32P]ATP in vitro. We have analyzed the different sites in MAP-2 phosphorylated by these three kinases in fresh or boiled cytosol from different regions of the rat brain, in particular the olfactory bulb, where only one form (MAP-2B) is present, and the cerebral cortex, where both forms (MAP-2A and MAP-2B) are equally enriched. Cyclic AMP dependent protein kinase and calcium/calmodulin-dependent protein kinase II phosphorylated four common phosphorylation sites, as well as a number of distinct sites that were unique to each enzyme. Calcium/phospholipid-dependent protein kinase phosphorylated a minimum of 15 sites, only one of which appeared to be shared with the other protein kinases. Only serine residues were phosphorylated by cyclic AMP-dependent and calcium/phospholipid-dependent protein kinases, while both serine and threonine residues were phosphorylated by calcium/calmodulin dependent protein kinase II. No differences were observed in the peptide maps of phospho-MAP-2 prepared from different brain regions. These results emphasize the complexity of the phosphorylation systems that may regulate the function of MAP-2 in situ. PMID- 2561877 TI - Image localized 31P magnetic resonance spectroscopy of the human liver. AB - Image localized 31P magnetic resonance (MR) spectroscopy of the liver was performed in twelve normal volunteers and seven patients with hepatic tumours. The tumours which were clearly imaged by proton MR could also be distinguished from normal tissue because of spectral differences. The malignant tumours had significantly elevated phosphomonoester/inorganic phosphate and phosphomonoester/beta-adenosine triphosphate ratios, probably due to elevated tumour concentrations of phosphocholine and phosphoethanolamine, which are intermediates in the synthesis of membrane phospholipids. The pH values of the malignant tumours were elevated compared to normal hepatic parenchyma. Liver spectra in two patients with the commonest benign hepatic neoplasm, cavernous haemangioma, differed from both normal tissue and the malignant tumours in having a very low signal/noise ratio but apparently normal relative levels of phosphomonoester. PMID- 2561878 TI - AZT/ddC combination may benefit AIDS sufferers. PMID- 2561876 TI - Purification and characterization of calmodulin-stimulated protein kinase II from two-day and adult chicken forebrain. AB - Soluble calmodulin-stimulated protein kinase II has been purified from 2-day and adult chicken forebrain. At both ages the holoenzyme eluted from a Superose-6B column with an apparent molecular weight of approximately 700,000 daltons and contained three subunits. The subunits were found to be the counterparts of the alpha, beta, and beta' subunits of the enzyme purified from adult rat brain in that they had one-dimensional phosphopeptide maps that were indistinguishable from those of the corresponding subunit in the rat enzyme and they migrated in SDS-polyacrylamide gels with the same apparent molecular weights. However, the doublet formed by the beta subunit was much more clearly resolved in the chicken enzyme and the beta' subunit, which was much more abundant in the adult chicken than in the adult rat, was also found to be a doublet. The ratio of the concentrations of the alpha and beta subunits changed during development. By autoradiography following autophosphorylation, the alpha:beta ratios of the 2-day and adult enzymes were 0.89 +/- 0.07 and 1.92 +/- 0.26, respectively; by silver staining the alpha:beta ratios were 0.95 +/- 0.11 and 1.85 +/- 0.17, respectively. The concentration of the beta' subunit was equal to that of the beta subunit at both ages. Autophosphorylation produced a decrease in the electrophoretic mobility of the alpha and beta subunits in SDS-polyacrylamide gels and a marked decrease in the calcium dependence of the substrate phosphorylation activity of the enzyme at both ages. The purified enzyme from chicken brain appeared to be more stable under standard in vitro assay conditions than the rat enzyme, and this was particularly so for the enzyme from 2-day forebrain. PMID- 2561879 TI - Fluconazole found effective against candidiasis. PMID- 2561880 TI - Noninvasive breast cancer: Part 1. Ductal carcinoma in situ--incidence, presentation, guidelines to treatment. AB - In situ breast cancer, both ductal and lobular, is being diagnosed more often today because of improved screening programs and more sensitive mammography. Unfortunately, treatment options in the disease are based on retrospective studies with the natural history obscured. While considered a noninvasive cancer, it has been theorized that ductal carcinoma in situ is a preinvasive stage that will eventually evolve into an invasive cancer. The author reviews results of studies comparing treatment methods, including wide local excision, mastectomy, and radiation, and concludes that prospective studies must be performed before optimal therapy can be determined. PMID- 2561881 TI - Noninvasive breast cancer: Part 2. Lobular carcinoma in situ--incidence, presentation, guidelines to treatment. AB - Many feel that lobular carcinoma in situ is a marker for future development of invasive cancer in other areas of the breast. There also seems to be a substantial incidence of bilaterality in LCIS. Mammography is of limited value in the diagnosis because it cannot distinguish between LCIS and benign disease. Treatment is by no means clearcut. The author surveys five options, and outlines her own usual course of action. PMID- 2561882 TI - Human papilloma virus clinic treats couples. PMID- 2561883 TI - Fast and accurate test for human papilloma virus. PMID- 2561884 TI - [Tasks and aims of the after care of cancer patients in the hospital]. PMID- 2561885 TI - [Bilateral white lesions of the tongue. Description of a case]. PMID- 2561886 TI - [Granular CHA in oral surgery. Clinical and therapeutic aspects]. AB - The Authors report their experience and the clinical results obtained by the use of powders of calcium-hidroxylapatite (Dac Blu) as filling material for bone defects in many different cases. The positive biological response justifies a wider utilisation of this material in oral surgery. PMID- 2561887 TI - The sensitivity of alpha-adrenoceptors in nucleus tractus solitarii, anterior hypothalamus and hippocampus of spontaneous and renal hypertensive rats. AB - The responsiveness of the endogenous type II inhibitor of protein kinases to clonidine was used as an index of the reactivity of alpha 2-adrenoceptors in the brain. The stimulation of postsynaptic alpha 2-adrenoceptors by clonidine resulted in a dose-dependent decrease in type II inhibitor activity. In the nucleus tractus solitarii and anterior hypothalamus of spontaneously hypertensive rats (SHR) and renal hypertensive rats (RHR) the clonidine-induced decrease in type II inhibitor activity was markedly reduced. In contrast, clonidine was equipotent in reducing type II inhibitor activity in the hippocampus of those rats and in control normotensive animals. In addition to that, in SHR and RHR the hypothermic activity of clonidine was not reduced. Thus, it seems that in experimental hypertension the decrease of the reactivity of alpha 2-adrenoceptors occurs in the brain structures responsible for the blood pressure regulation. PMID- 2561888 TI - Neurochemical correlates of differences in responses to psychotropic drugs. I. Apomorphine and morphine effects on locomotor activity of C57/BL and Balb/C mice. AB - C57/BL mice have higher basal locomotor activity than Balb/C mice and react to apomorphine (4 mg/kg) and morphine (20 mg/kg) with motor stimulation, while in Balb/C mice this dose of apomorphine depressed the basal locomotor activity and morphine did not affect it. No differences in the characteristics of opiate mu and delta receptors in the whole brain of mice of both strains were found, but the responsiveness of the central alpha-adrenoceptor system, as measured with the accumulation of inositol phosphate in brain slices after exposure to noradrenaline was different: the low concentrations of noradrenaline (5 microM) produced stronger effect in Balb/C mice, but the maximum responses tended to be stronger in C57/BL mice. PMID- 2561889 TI - Antidepressant activities of WEB 1881, a new nootropic agent. AB - The central action, particularly potential antidepressant activity of WEB 1881, a new nootropic drug related to piracetam, was investigated in rats and mice. WEB 1881 antagonizes the reserpine- and apomorphine-induced hypothermia, potentiates the behavioral effect of DOPA and dihydroxyphenylserine, as well as the TRH induced hyperthermia. Piracetam was only effective in the reserpine and DOPA tests. WEB 1881 is inactive in immobility test of Porsolt et al. It enhances the hind limb flexor reflex of the spinal rat, this effect being antagonized by prazosin and cyproheptadine. It exerts no effect on head twitches induced by L-5 hydroxytryptophan. The studied compound increases the noradrenaline and dopamine and turnover in the forebrain and brain stem. WEB 1881 given repeatedly potentiates the clonidine-induced aggressiveness and has no effect on the locomotor hyperactivity induced by D-amphetamine. The results indicate that in a number of tests WEB 1881 acts like other antidepressant drugs (but in others not), moreover, they suggest that this action is--at least partly--mediated by the central noradrenaline system. PMID- 2561890 TI - Oxygen radicals stimulate gallbladder glycoprotein secretion. AB - Mucin hypersecretion in the gallbladder is thought to be a key factor in the nucleation of cholesterol gallstones. In this study, we evaluated the effect of several oxygen radical-generating systems on glycoprotein release from guinea pig gallbladder explants. Hydroxyl radicals (OH.) released by hypoxanthine-xanthine oxidase or FeCl3-ascorbate caused a significant increase in glycoprotein secretion from gallbladder explants pre-incubated with [3H] glucosamine. Human neutrophils activated with f-Met-Leu-Phe, a chemotactic peptide, released oxygen radicals that also stimulated gallbladder glycoprotein release. The mechanism of this effect is most probably related to perturbation of lipid membrane structure by the oxygen radicals with subsequent activation of glycoprotein release. PMID- 2561891 TI - Effects of forskolin on the longitudinal internal resistance (ri) in rabbit sinus node strips. AB - The effect of several concentrations of 7-0-hemisuccinyl, 7-deacetyl forskolin on ri was studied by means of microelectrode and single sucrose gap techniques. Twenty mumol.l-1 of forskolin added to the sucrose gap lowered ri by 15% below the control value. The drug applied at 50 mumol.l-1 initially decreased ri by 31%, and then it increased the resistance by 43% above the control value. The possible mechanisms underlying these effects are discussed in terms of cAMP and Ca action on the cell coupling. PMID- 2561892 TI - Acid-base values in the blood of the European bison Bison bonasus (L.). PMID- 2561893 TI - Clinical values for an index predicting postoperative residual liver function by pre-operative liver-scintigraphy in patients with liver disease. AB - Hepatic resection is essential in treating hepatocellular carcinoma. However, before an operation, it is difficult to predict the functional reserve in the remnant following massive resection. We devised an original method by which effective liver volume was measured by liver scintigraphy. In order to predict the residual liver function before hepatic resection in a preoperative radiocolloid study, we obtained a predictive index by combining the K values with effective liver volumes which seemed to have the estimated residual liver function. Twenty-one patients with liver or biliary tract disease were selected at random for the present study. We divided them into 3 groups in accordance with prognosis after hepatic resection. There were statistically significant difference between the deceased group who died from hepatic failure and the group who died from causes other than hepatic failure; and between the deceased group who died from hepatic failure and the living group in the preictive index (p less than 0.01). Our data suggest that if the predictive index is above 0.45, the probability of hepatic failure after hepatic resection is low. We concluded that our predictive index is useful to use in preoperative prediction of post hepatectomic residual liver function. PMID- 2561894 TI - Thyroid carcinoma in solitary hot thyroid lesions on Tc-99m sodium pertechnetate scans. AB - Sixteen patients with nonsuppressible solitary hot thyroid lesions (SHTL) identified on T3 suppression images using Tc-99m sodium pertechnetate were studied over a period of 5 years. Of the 16 patients, 7 (44%) had papillary adenocarcinoma (PAC) and 9 (56%) had follicular adenoma (FA). Of the 7 patients with PAC, 3 were toxic and 4 nontoxic. Of the 9 patients with FA, 2 were toxic and 7 nontoxic. The Tl-201 chloride thyroid scans were useful in locating SHTL and revealing extranodular thyroid tissue. The echography was sensitive to visualization of the nodule structures. However, there were no significant differences between the clinical findings, radionuclide images, and echograms between for PAC and FA. All patients with PAC were treated by partial thyroidectomy and there were neither regional nor distant metastasis in any of them. In conclusion, our study provided the following extremely interesting result: SHTL in the present series have a higher incidence of malignancy than previously reported autonomously functioning thyroid lesions (AFTL). Histological examination is necessary for the diagnosis and management of SHTL and surgical treatment should be considered. PMID- 2561895 TI - SPECT images using 99mTc(V)-DMS in lung metastasis of osteosarcoma. AB - Color images of single-photon emission computed tomography (SPECT) using technetium-99m(V) dimercaptosuccinic acid (99mTc(V)-DMS) were demonstrated in a patient with lung metastases from osteosarcoma. SPECT image using 99mTc(V)-DMS could be useful for the detection of lung metastasis from osteosarcoma. PMID- 2561898 TI - Radiology of the pancreas. PMID- 2561896 TI - Synthesis and evaluation of 11C-PK 11195 for in vivo study of peripheral-type benzodiazepine receptors using positron emission tomography. AB - The biodistribution of 3H-PK 11195, an antagonist of the peripheral-type benzodiazepine receptors, was studied in mice. High accumulations of radioactivity in the heart, lung, spleen, kidney and adrenal were observed after intravenous injection of tracer amounts of 3H-PK 11195 into the mice. The radioactivity in the heart, lung, spleen, kidney and adrenal was significantly decreased by the coadministration of carrier PK 11195, which indicated that PK 11195 specifically binds to the receptors. No radioactive metabolites were observed in the heart, lung and brain 20 min after intravenous administration of 3H-PK 11195. The accumulation of 3H-PK 11195 in the lung was not affected by pretreatment with either alpha-methyl benzylamine or imipramine, suggesting that 3H-PK 11195 specifically binds to the receptors. The ratios of radioactivity of the kidney, adrenal and spleen to blood increased as a function of time, whereas that of the lung and heart rapidly reached to a steady state. 11C-PK 11195 was synthesized by the N-methylation of desmethyl precursor yielding more than 100 mCi with high specific activity (more than 1.4 Ci/mumol). The labeling and purification procedure was completed within 23 min after the end of bombardment (EOB). The 11C-PK 11195 solution for injection seems to have a high potential for the in vivo study of the peripheral-type benzodiazepine receptors in the living human by means of positron emission tomography (PET). PMID- 2561899 TI - Bicarbonate inhibition of Saccharomyces cerevisiae and Hansenula wingei growth in apple juice. AB - The ability of sodium bicarbonate to inhibit growth of Saccharomyces cerevisiae and Hansenula wingei in apple juice was investigated. Sodium bicarbonate at concentrations of 0.06, 0.12, and 0.24 M was added to pasteurized apple juice that was then inoculated with 10(3) or 10(5) cfu/ml of either yeast. Growth of both yeasts was inhibited by 0.12 M sodium bicarbonate when incubation was at 4 degrees C; 0.24 M sodium bicarbonate caused a slow die off of yeast. At 18 degrees C, H. wingei became more sensitive and died in the presence of 0.12 M sodium bicarbonate, but S. cerevisiae became resistant to 0.24 M sodium bicarbonate. These results could not be attributed to bicarbonate-induced pH elevation or sodium. Potassium and ammonium bicarbonate were also inhibitory, implicating bicarbonate ion as the antimicrobial agent. PMID- 2561897 TI - Radionuclide venography of lower limbs by subcutaneous injection: comparison with venography by intravenous injection. AB - We have proved that subcutaneous injection (SC) of a small dose of Tc-99m pertechnetate (1 to 2 mCi: 37 to 74 MBq) at acupuncture points (K-3 and B-60) may offer an alternative method of radionuclide venography (RNV) of the lower limbs. In this study, we compared intravenous (IV) RNV and SC-RNV in 22 consecutive cases with typical signs and symptoms suggesting venous abnormality of the lower limb(s) from March to May 1988. They are 11 male and 11 female, aged 47.7 +/- 15.7 years. Among the 44 limbs of the 22 cases, 4 were normal, 12 (27.3%) were found to have varicose veins in the legs only, 18 (40.9%) had partial stenosis of the deep veins (14 poplito-tibial and 4 superficial femoral), and 13 (29.6%) had complete stenosis of the deep veins (4 poplitotibial, 1 superficial femoral and 8 ilio-femoral. SC-RNV showed almost the same results as IV-RNV in 21 (47.7%), superior to IV-RNV in 22 (50%) (including 4.6% failure of IV-RNV), and inferior to IV-RNV in 1 (2.3%). We conclude that SC-RNV is definitely an alternative method of lower-limb venography. Since it is in most cases superior to IV-RNV, we suggest that it can take the place of IV-RNV in routine work. PMID- 2561900 TI - Preincubation time and the use of oxygen indicators in determining the microbiological quality of aseptically packed pea and tomato soup. AB - In order to get accurate information about the preincubation time needed for viscous aseptic products, the growth characteristics of three food poisoning organisms Staphylococcus aureus, Clostridium perfringens and Bacillus cereus in pea soup, and one spoilage organism, Lactobacillus plantarum, in tomato soup, were followed during a preincubation period at 30 degrees C for 14 days. The sterile food packs were sealed into polyamidepolyethylene laminate bags containing different headspace gas concentrations (21% oxygen + 80% nitrogen, 5% oxygen + 95% nitrogen and 100% nitrogen). Bacterial growth was followed every second day by counting the number of colony forming units and following the headspace oxygen and carbon dioxide concentrations. The bacterial strains differed in their growth characteristics, especially in their ability to consume oxygen from the headspace, and also in their ability to produce carbon dioxide. The headspace oxygen concentration was also followed by oxygen indicators in order to determine the correlation with the oxygen concentration of the headspace and the growth of bacteria. The bacterial growth followed more closely the concentration of carbon dioxide than the concentration of oxygen. It was concluded that further studies are needed to determine the preincubation period for viscous aseptic products in order to furnish accurate data on the growth characteristics of low numbers of bacteria in aseptic foods at different headspace gas compositions. PMID- 2561901 TI - Several new AIDS drugs being tested. PMID- 2561902 TI - Intraductal noninvasive breast cancer: a comparison of three local treatments. AB - In selecting appropriate local treatment for intraductal noninvasive breast cancer, the following factors should be considered: Method of detection (palpable mass vs mammographic finding only vs incidental finding at biopsy for a benign lesion), the size and extent of the tumor, margins of resection, and the histologic pattern. Mastectomy is recommended for extensive tumors, and for patients with gross multicentric disease. Conservative surgery and radiation is an alternative for patients with focal lesions, negative margins of resection, and no evidence of residual microcalcifications on a post-biopsy mammogram. Consideration of wide excision alone should be restricted to those with occult primary tumors less than or equal to 2.5 cm in size, detected by mammographic microcalcifications only, and where complete excision is confirmed. The histologic subtype or nuclear grade may be especially important in evaluating patients for wide excision alone. PMID- 2561903 TI - Sodium periodate treatment modulates the accessory and regulatory functions of alveolar macrophages in T-cell responses. AB - It is known that murine alveolar macrophages function inefficiently as antigen presenting cells for the in vitro activation of macrophage-depleted T-lymphocyte populations and that they suppress antigen-stimulated lymphoproliferative responses in a dose-dependent manner. The present studies were carried out to determine whether oxidation of the alveolar macrophage surface could alter these activities. Viable alveolar macrophages were treated with varying concentrations of sodium periodate then cultured with either unfractionated or adherent cell depleted lymphocyte populations obtained from BCG-immunized animals and challenged with PPD. It was demonstrated that moderate oxidation of the alveolar macrophage surface with sodium periodate abrogated their ability to suppress antigen-stimulated proliferation of unfractionated lymphocyte populations, as well as resulted in initiation of lymphoproliferation in an antigen-stimulated, adherent cell-depleted spleen cell populations. It is conceivable that mild oxidation of the alveolar macrophage membrane results in a more favorable interaction between the alveolar macrophage and responding T-lymphocytes, thereby reversing the alveolar macrophage-mediated suppression of antigen-stimulated lymphoproliferation and allowing for effective antigen-presenting function of these cells. PMID- 2561904 TI - Pathogenesis of diabetic neuropathy: role of altered phosphoinositide metabolism. AB - A unifying metabolic hypothesis completely accounting for the development of one or more of the chronic complications of diabetes on the basis of a single aspect of disturbed glucose metabolism resulting from insulin deficiency and/or hyperglycemia has been sought by clinical and basic scientists for decades. A growing body of loosely related but internally consistent scientific data obtained from cultured cells, incubated tissue preparations, animal models, and man implicate sorbitol- and glucose-induced myo-inositol depletion and altered phosphoinositide metabolism in a series of secondary biochemical, functional, and architectural abnormalities in the PNS in diabetes. These early metabolically based functional and structural changes simulate those that characterize human diabetic neuropathy. Can abnormal phosphoinositide metabolism in diabetic nerve thereby by itself explain the development of chronic diabetic neuropathy with all of its clinical complexity and heterogeneity? Almost certainly not. Even if the entire contribution of hyperglycemia to the development of diabetic neuropathy were mediated by secondary abnormalities in phosphoinositide metabolism, other factors must also play a role. Witness the differences in the histopathological picture of neuropathy in patients with IDDM and NIDDM despite similar durations and severity of diabetes, the apparent influence of age and gender on the appearance of early neuropathy in patients with IDDM, and the association of alcohol consumption with diabetic neuropathy. While early metabolic and functional disturbances in diabetic nerve such as impaired (Na,K)-ATPase function and paranodal swelling are empirically attributable to abnormal myo-inositol and phosphoinositide metabolism, more advanced abnormalities such as axo-glial dysjunction may reflect superimposed independent biochemical and/or hormonal defects (although, as mentioned previously, aldose reductase inhibition decreases axo-glial dysjunction in diabetic humans). The PNS has only a limited repertoire of responses to a variety of insults, so that Wallerian degeneration, axonal atrophy, impaired axonal transport, and dystrophic changes in diabetic neuropathy may represent multiple factors. On the other hand, the increasingly recognized importance of the phosphoinositide cascade in neuromodulation may attribute a progressively wider range of disturbances in the diabetic PNS to myo-inositol depletion and associated defects in phosphoinositide metabolism. Thus, while all effects of aldose reductase inhibitors in the PNS of diabetic rats have been reproduced by myo-inositol supplementation when this alternative intervention has been tested, the exact role of phosphoinositide metabolism in most of these responses is not well understood.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2561905 TI - Radiation treatment of brain tumors: concepts and strategies. AB - Ionizing radiation has demonstrated clinical value for a multitude of CNS tumors. Application of the different physical modalities available has made it possible for the radiotherapist to concentrate the radiation in the region of the tumor with relative sparing of the surrounding normal tissues. Correlation of radiation dose with effect on cranial soft tissues, normal brain, and tumor has shown increasing effect with increasing dose. By using different physical modalities to alter the distribution of radiation dose, it is possible to increase the dose to the tumor and reduce the dose to the normal tissues. Alteration of the volume irradiated and the dose delivered to cranial soft tissues, normal brain, and tumor are strategies that have been effective in improving survival and decreasing complications. The quest for therapeutic gain using hyperbaric oxygen, neutrons, radiation sensitizers, chemotherapeutic agents, and BNCT has met with limited success. Both neoplastic and normal cells are affected simultaneously by all modalities of treatment, including ionizing radiation. Consequently, one is unable to totally depopulate a tumor without irreversibly damaging the normal tissues. In the case of radiation, it is the brain that limits delivery of curative doses, and in the case of chemical additives, it is other organ systems, such as bone marrow, liver, lung, kidneys, and peripheral nerves. Thus, the major obstacle in the treatment of malignant gliomas is our inability to preferentially affect the tumor with the modalities available. Until it is possible to directly target the neoplastic cell without affecting so many of the adjacent normal cells, the quest for therapeutic gain will go unrealized. PMID- 2561907 TI - [Malignant pleomorphic fibrous histiocytoma of the lung. A clinical case]. AB - A 44 year old woman presented with a bloody sputum, associated to bilateral lung nodules resembling metastatic lung carcinoma. The diagnosis of pulmonary malignant pleomorphic fibrous histiocytoma was made through an open lung biopsy. Histological considerations and a literature review related to this rare condition are induced. PMID- 2561906 TI - [Lactic acidosis in diabetic patients associated with buformin]. AB - We report two diabetic patients who developed lactic acidosis following the use of Buformin. Treatment consisted of mechanical ventilation, massive bicarbonate administration, circulatory support with dopamine and peritoneal dialysis. Despite this, both patients died. PMID- 2561908 TI - [Stress induced change in ACTH plasma levels in rats treated with benzodiazepine derivatives]. PMID- 2561909 TI - [Effect of calcium antagonists on the metabolism of phosphoinositides in isolated liver cells]. PMID- 2561910 TI - AIDS-associated Kaposi's sarcoma: a molecular model for its pathogenesis. AB - Kaposi's sarcoma is a mesenchymal tumor that is frequently observed in AIDS patients. Although the tumor is clearly associated with HIV-1 infection, little is known about the molecular events underlying its pathogenesis. Here we consider the potential roles of specific cytokines and the virally encoded tat product in inducing the tumor. PMID- 2561911 TI - ADP-ribosylation of thylakoid membrane polypeptides by cholera toxin is correlated with inhibition of thylakoid GTPase activity and protein phosphorylation. AB - Incubation of pea thylakoid membranes with [32P]-NAD+ in the presence of cholera toxin resulted in the [32P]-ADP-ribosylation of a 60 kDa thylakoid membrane polypeptide. When ATP was included in the incubation mixture, a 29 kDa polypeptide was also labelled. In the absence of electron transfer cofactors or inhibitors, the extent of labelling depended on whether the membranes were preincubated in the light or dark and also on the developmental stage of the leaves used for thylakoid isolation. Irrespective of the latter, the strongest labelling was observed when DCMU was present in the light. After pretreatment of the thylakoid membranes with cholera toxin plus NAD+ under the same conditions, light-stimulated GTPase activity and protein phosphorylation were inhibited. The extent of inhibition for both processes appeared to be correlated with the amount of [32P]-ADP-ribosylation found when [32P]-NAD+ was included in the pretreatment mixture. The data presented are fully consistent with the 60 and 29 kDa polypeptides functioning as thylakoid membrane associated guanine nucleotide binding regulatory proteins. PMID- 2561912 TI - Vasoactive intestinal peptide receptor regulation of cAMP accumulation and glycogen hydrolysis in the human Ewing's sarcoma cell line WE-68. AB - This study describes functional characteristics of receptors for vasoactive intestinal peptide (VIP) on human Ewing's sarcoma WE-68 cells. These characteristics include 125I-VIP binding capacity, cellular cAMP generation, glycogen hydrolysis, and pharmacological specificity. Binding studies with 125I VIP showed specific, saturable, binding sites for VIP in WE-68 cells. Scatchard analysis revealed the presence of a single class of high-affinity binding sites that exhibited a dissociation constant (Kd) of 90 pM and a maximal binding capacity (Bmax) of 24 fmol/mg of protein. VIP and VIP-related peptides competed for 125I-VIP binding in the following order of potency: human (h) VIP greater than human peptide with N-terminal histidine and C-terminal methionine (PHM) greater than chicken secretin much greater than porcine secretin. Glucagon and the C-terminal fragments VIP[10-28] and VIP[16-28] and the VIP analogue (D Phe2)VIP did not inhibit 125I-VIP binding. Addition of hVIP to WE-68 cells provoked marked stimulation of cAMP accumulation, hVIP stimulated increases in cAMP content were rapid, concentration-dependent, and potentiated by 3-isobutyl-l methylxanthine (IBMX). Half-maximal stimulation (EC50) occurred at 150 nM hVIP. The ability of hVIP and analogues to stimulate cAMP generation paralleled their potencies in displacing 125I-VIP binding. (D-Phe2)VIP, VIP[10-28], VIP[16-28], and (p-Cl-D-Phe6, Leu17)VIP, a putative VIP receptor antagonist, affected neither basal cAMP levels nor hVIP-induced cAMP accumulation. WE-68 cell responses to hVIP were desensitized by prior exposure to hVIP. Desensitization to hVIP did not modify the cAMP response to beta-adrenergic stimulation, and beta-adrenergic agonist desensitization did not modify responses to hVIP. hVIP also induced a time- and concentration-dependent hydrolysis of 3H-glycogen newly formed from 3H glucose in WE-68 cultures. hVIP maximally decreased 3H-glycogen content by 36% with an EC50 value of about 8 nM. The order of potency of structurally related peptides of hVIP for stimulation of glycogenolysis correlated with their order of potency for inhibition of 125I-VIP binding. IBMX potentiated the glycogenolytic action of hVIP and PHM. The simultaneous presence of the calcium channel antagonist verapamil or the calcium ionophore A 23187 did not influence the glycogenolytic and cAMP stimulatory effects of hVIP. Collectively, these data indicate that Ewing's sarcoma (WE-68) cells are endowed with genuine VIP receptors which are coupled to the formation of cAMP that probably serves a second messenger role in stimulating glycogen hydrolysis in these cells in response to VIP.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2561913 TI - Epinephrine potentiates calcium mobilization and activation of protein kinases in platelets stimulated by ADP through a mechanism unrelated to phospholipase C. AB - ADP, added to suspensions of aspirinized 32P-prelabelled washed platelets, induced reversible platelet aggregation, the rapid elevation of cytosolic Ca2+ (maximum at 2 s), 20 kDa myosin light chain phosphorylation (maximum faster than 3 s), 40 kDa protein phosphorylation (maximum at 3-10 s) and phosphatidic acid formation (maximum at 30 s). Prior addition of epinephrine potentiated platelet aggregation, cytosolic Ca2(+)-elevation, 20 and 40 kDa protein phosphorylation evoked by ADP, but it did not enhance phosphatidic acid formation induced by ADP. The potentiating effect of epinephrine on aggregation, cytosolic Ca2(+)-increase and 20 and 40 kDa protein phosphorylation induced by ADP was also observed in the presence of EGTA. Ethylisopropylamiloride, an inhibitor of Na+/H(+)-exchange, did not affect the potentiation of ADP-induced platelet aggregation by epinephrine. We conclude that epinephrine primes platelets to increase Ca2(+)-influx and Ca2(+)-mobilization in response to ADP. The potentiation of cytosolic Ca2(+) elevation by epinephrine leads to further stimulation of myosin light chain phosphorylation and protein kinase C activation and ultimately to enhanced platelet aggregation. These effects of epinephrine do not seem to take place at the level of phospholipase C. PMID- 2561915 TI - Actinomycin D-induced hepatotoxicity. PMID- 2561914 TI - Fibroblast growth factor potentiates receptor- and nonreceptor-mediated stimulation of adenylate cyclase in hamster fibroblasts. AB - Basic fibroblast growth factor (FGF) has no effect alone on the basal cAMP synthesis in Chinese hamster fibroblasts (CCL39) but it potentiates (by up to 50%) the stimulation of adenylate cyclase by prostaglandin E1, cholera toxin or forskolin. This potentiating effect is not abolished by pretreatment of the cells with pertussis toxin, which indicates that it is not due to the withdrawal of a tonic inhibition of adenylate cyclase by the pertussis toxin-sensitive inhibitory GTP-binding protein (Gi). Therefore, we conclude that FGF enhances the activation of adenylate cyclase by the stimulatory GTP-binding protein (Gs). Although activation of protein kinase C in CCL39 cells results in a similar potentiation of cAMP production, we provide evidence that the effect of FGF is not mediated by protein kinase C, since (1) the potentiating effects of FGF and phorbol esters are additive and (2) FGF effect persists after down-regulation of protein kinase C. A role of FGF-induced rise in cytoplasmic Ca2+ can also be ruled out because the FGF effect is not mimicked by a Ca2+ ionophore and it persists in Ca2(+)-free medium. Since a similar potentiating effect on cAMP production is elicited by epidermal growth factor, a mitogen known to activate a receptor tyrosine kinase, we suggest that the FGF effect on adenylate cyclase might be mediated by the tyrosine kinase activity that is very likely to be associated with FGF receptors. PMID- 2561916 TI - Itraconazole versus ketoconazole for the prophylaxis of fungal infection in neutropenic children: results of two consecutive nonrandomized studies. AB - Two consecutive nonrandomized studies were conducted in children with prolonged granulocytopenia to evaluate the prophylactic antifungal activity of ketoconazole and the new triazole itraconazole. The conditions were equivalent in both studies. The incidence of colonization was 10% in the ketoconazole group and 19% in the itraconazole group (this difference is not significant). For suspected and proven infections, the incidence was 5% for ketoconazole and 10% for itraconazole, but the incidences were too low for statistical comparison. Although the colonization rate was higher for itraconazole, there was no sign of Aspergillus, whereas in the ketoconazole group an autopsy, proven aspergillosis has been reported. PMID- 2561917 TI - Wilms' tumor complicated by veno-occlusive disease of the liver (VOD): current concepts and a case report. PMID- 2561918 TI - Genital papillomavirus infection. PMID- 2561919 TI - Liver transplantation for malignant tumours. AB - Despite considerable overall progress in human liver transplantation the results obtained in patients with malignant tumours have not improved significantly over the past years. One of the crucial questions in the ongoing controversial discussion remains the identification of tumour patients with the most favourable prognosis. In a consecutive series of 114 patients who received hepatic transplants for various malignant tumours of the liver and biliary tract, at least some factors could be shown to play a prognostic role. Regarding the histological type of tumour, fibrolamellar carcinoma, epitheloid haemangioendothelioma, and endocrine hepatic metastases seem to have a better long-term survival, whereas cholangiocellular carcinoma and other liver metastases had the worst outcome. In patients with primary liver or proximal bile duct cancer there was a significant influence of the pathological tumour stage at the time of transplantation: significant palliation or cure was almost essentially restricted to patients with early tumour stages as compared with advanced primary tumours and extrahepatic spread where early tumour recurrence developed in all recipients. Thus, the present concept for the treatment of malignant hepatobiliary tumours should include partial as well as total hepatectomy with subsequent liver replacement. In cases of non-resectable lesions or intrahepatic tumour recurrence following previous resection, liver transplantation offers the only chance for long-term survival. PMID- 2561920 TI - [Cystic mucinous neoplasms of the pancreas]. AB - The Authors report on three cases of Mucinous Cystic Neoplasms of the pancreas. These tumours are rare and the diagnosis is based on echography and CT scanning. After a radical excision the prognosis is good although these tumours are "all" potentially malignant. PMID- 2561921 TI - Some epidemiological observations in encephalitis in children admitted to the Burdwan Medical College Hospital, Burdwan, West Bengal. PMID- 2561922 TI - [Clinical diagnosis and pathogenesis of myocardial infarction complicated by hypertrophic cardiomyopathy: review of eight cases]. AB - Among 144 patients with hypertrophic cardiomyopathy, eight (58.3 +/- 7.0 years, M:F = 7:1) had complicating myocardial infarction, which was diagnosed clinically and by elevated cardiac enzymes or new Q-waves on electrocardiography. Coronary occlusion or stenosis evidenced by coronary angiography and nuclear cardiological findings were investigated. In six of the eight patients, coronary atherosclerosis caused infarction. These patients had many coronary risk factors compared to the other two patients. Sixteen of the 144 patients (11%) with hypertrophic cardiomyopathy had coronary atherosclerosis, the rate of which is reportedly 10 to 20%. Two of the eight patients had no coronary atherosclerosis. One patient had a diffusely spastic diathesis provoked by the intravenous administration of ergonovine maleate during coronary angiography, suggesting that coronary spasm caused myocardial infarction. The other patient had recurrent episodes of supraventricular tachyarrhythmia and no evidence of spasm during coronary angiography, suggesting coronary embolism as a cause of myocardial infarction. Myocardial infarction in patients with hypertrophic cardiomyopathy and normal coronary arteries as advocated by Maron et al. may have such pathogenesis. We conclude that coronary angiography may be mandatory in patients with hypertrophic cardiomyopathy, especially those who have many coronary risk factors and anginal symptoms. In these patients, ST-T changes and abnormal Q waves on electrocardiography sometimes may be misleading when diagnosing the occurrence of acute myocardial infarction by electrocardiography alone. In such cases, infarct-avid scintigraphy with 99 m-Tc pyrophosphate is preferable. PMID- 2561923 TI - Alpha-2-adrenoreceptor binding as a possible vulnerability marker for affective disorders. AB - The affinity (1/Kd) and density (Bmax) of alpha 2-adrenoreceptors in platelet membranes were studied in patients with major depressed illness (n = 10), affected first-degree relatives (n = 17), nonaffected first-degree relatives (n = 44) and controls (n = 31). The alpha 2 selective antagonist 3H-yohimbine was used as the radioligand. The mean Bmax values of affected subjects (probands and relatives) were significantly lower than those of controls. There was no difference in Kd values between the controls and affected subjects. There was a positive gradient of the mean Bmax values from the groups of probands to affected relatives, unaffected relatives and control subjects. A familial effect of Bmax values between members of the same families confirms a genetic control of alpha receptor affinity. These results support the hypothesis that the density of alpha 2-adrenoreceptors, evaluated by 3H-yohimbine binding on human platelets, could be a potential vulnerability marker for affective disorder. PMID- 2561924 TI - [Comparison of the susceptibility of Candida strains to ketoconazole and itraconazole in in vitro experiments]. AB - The activity of ketoconazole and itraconazole against 102 strains of Candida sp. was compared in the investigation "in vitro". The investigational material was taken from patients with different types of mycosis. The values of MICs of ketoconazole and itraconazole were 0.04-100.0 micrograms/ml and 0.02-35.0 micrograms/ml, respectively. From the data obtained it is evident that the average efficacy of itraconazole is 3 to 10 times higher in comparison with ketoconazole though 8 strains (7, 8 percent) of investigated strains have shown greater sensitivity to ketoconazole. PMID- 2561925 TI - [Antifungal activity of itraconazole and ketoconazole investigated in vitro, determined with minimal inhibitory concentrations and mycelial cell transformation in Candida albicans]. AB - Results of studies "in vitro" of the susceptibility of 82 strains of Candida albicans for itraconazole and ketoconazole have been report. The strains were isolated from lesions of the mucous membranes and skin. The susceptibility of Candida albicans was tested to Minimal Inhibitory Concentration (MIC) and Total Inhibition Germination Tubes (TI100). The values of MICs and TI100 of itraconazole were 0.02-35.0 micrograms/ml and 0.02-18.0 micrograms/ml, respectively. The similar values of MICs and TI100 of ketoconazole were 0.04-80.0 micrograms/ml and 0.04-20.0 micrograms/ml. It has been showed that TI100 can be used as preliminary and rapid test of Candida albicans susceptibility for these drugs. PMID- 2561926 TI - [Cutaneous cylindroma--cylindroma cutis]. AB - A case of disseminated cylindroma of the scalp is reported. Big tuberous lesions were removed surgically, and small nodules were treated twice with liquid nitrogen using a UK-32 device for cryotherapy. PMID- 2561927 TI - Lipolysis by corticotropin in fat cells from hypothyroid rats. Effect of adenosine deaminase. AB - Thyroid hormones are required for maximal stimulation of lipolysis of fat cells by catecholamines corticotropin and glucagon. Several reasons have been given to explain this fact, but all of them are controversial and still not definitive. It has been proposed that adenosine is an important factor in the low lipolytic response to catecholamines by fat cells of hypothyroid rats. This proposal has been studied with corticotropin. There has been no recuperation of maximal lipolysis when fat cells of hypothyroid rats were stimulated by corticotropin in the presence of adenosine deaminase. PMID- 2561928 TI - Pharmacology of acute alcoholic intoxication. AB - Research of the alcohol action mechanism on the SNC in acute alcoholic intoxication (AAI) has been dealt in various ways. On one side the alcohol action -apparently most unspecific--on cellular membranes has been studied. Other authors, instead, have studied more specific alcohol effects on three types of neurotransmitters: opioid peptides, GABA and catecholamines. The effect of alcohol on cellular membranes seems to be beyond any doubt. Alcohol action on specific neurotransmitters is the object of controversy, especially in the case of endogenous opioids. There are data which strongly support the participation of the GABA receptors in the AAI. Modifications produced in the cellular membrane by alcohol action can modify the structure of the function of the membrane receptors. On the other hand, distinct receptors may be localized in the same neuron, while the existence of interactions between different neurotransmitters is well known. Therefore, the various hypotheses previously stated are not mutually exclusive. PMID- 2561929 TI - A short introduction to positron emission tomography. PMID- 2561930 TI - Imaging mu-opiate receptors in epilepsy by positron emission tomography. PMID- 2561931 TI - Functional brain mapping with positron emission tomography. PMID- 2561932 TI - Active opioid binding protein from rat brain--a glycoprotein containing alpha methyl-D-mannoside residues. AB - Active opioid binding protein was partially purified from rat brain by Vicia bungei Ohwi lectin (VBL). Mannose also existed in opioid receptors since the binding between opioid binding protein and VBL is specific, via alpha-methyl-D mannoside (MeMan) residues. Since the active opioid binding protein purified by VBL chromatography was enriched about 200-folds. VBL can be used as effective purification tool which is much better than wheat germ agglutinin (WGA), a lectin commonly used in purifying opioid receptor. PMID- 2561933 TI - [Clonidine stimulates central nervous alpha 2 adrenoceptors not mediating Ca2+ channels]. AB - Seven renal hypertensive (2 kidneys--2 clipped) and 8 normotensive conscious dogs were given icv clonidine. After the medication the mean arterial blood pressure (MAP) were lowered and the heart rate (HR) and plasma norepinephrine (NE) were decreased. The magnitude of the decrease of MAP was greater than normotensive dogs; while no differences of decrease of NE was found. Nicardipine (10 micrograms/kg) icv caused significant increases of MAP and HR in both hypertensive and normotensive groups. The decrease of MAP induced by clonidine was not changed by the pretreatment of nicardipine icv in both groups. It is thus concluded that clonidine stimulates central nervous alpha adrenoceptors not mediating Ca2+ channels. PMID- 2561934 TI - [Phencyclidine receptors in porcine cerebral arteries]. AB - A specific, saturable, reversible, and selective binding site with Kd = 87 +/- 33 nmol/L, Bmax = 0.78 +/- 0.11 pmol/mg protein was detected in the binding of [3H] phencyclidine (PCP) to porcine cerebral blood vessels. Only ligands of PCP/sigma series were able to bind to the PCP receptors. [3H]PCP bound to its receptors was not displaced by etorphine or norepinephrine 0.1 mmol/L. A specific [3H]PCP binding site was found in porcine brain with Kd = 75 +/- 34 nmol/L, Bmax = 0.61 +/- 0.23 pmol/mg protein. Bioassay in vitro showed PCP enhanced the perfusion pressure of porcine cerebral blood vessels in a dose-dependent manner. This study provides direct evidence for PCP receptors on cerebral blood vessels, and suggests that PCP may produce cerebral vasospasm via PCP receptor interaction. PMID- 2561935 TI - [Effects of tetrandrine on cytoplasmic free calcium in rat neutrophils]. AB - Neutrophils (NP) cytoplasmic free Ca2+ concentrations ([Ca2+]i) under several conditions were measured with Quin 2 in rats. The process of loading Quin 2-AM into the cells followed by its hydrolysis was assured by monitoring the shift of fluorescent emission spectrum of Quin 2-AM to that of Quin 2. The resting [Ca2+]i of NP in Ca2(+)-containing and Ca2(+)-free solutions were 186 +/- 45 and 46 +/- 16 nmol/L, respectively, indicating extracellular calcium concentration ([Ca2+]0) plays an important role on [Ca2+]i. Among agents tested, prostaglandin E2 (PGE2) 100 nmol/L did not change [Ca2+]i significantly. Calcimycin 25 mumol/L, leukotriene B4(LTB4) 60 nmol/L and platelet activating factor (PAF) 10 nmol/L increased [Ca2+]i of NP in Ca2(+)-containing solution from 185 +/- 54, 175 +/- 36 and 188 +/- 54 to 814 +/- 67, 577 +/- 229 and 540 +/- 174 nmol/L, respectively. But, compound 48/80 3.2 micrograms/ml, LTB4 300 nmol/L, PAF 10 nmol/L and PAF 5 nmol/L plus LTB4 150 nmol/L did not change significantly [Ca2+]i of NP in Ca2(+) free solution, indicating that these agonists can not release intracellularly stored Ca2+ or no stored Ca2+ in NP is available. The rises of [Ca2+]i produced by calcimycin, PAF and LTB4 were markedly inhibited by tetrandrine (Tet) 65 mumol/L. These results, as a whole, show that Tet inhibits the rises of [Ca2+]i of NP induced by calcimycin, PAF and LTB4 via decreasing Ca2+ influx. The Ca2+ antagonism in this case may be related to its anti-allergic actions. PMID- 2561936 TI - Effect of fluorocarbon blood substitute on neutrophil phagocytic function. AB - Neutrophils were incubated at 37 degrees C for 2 h with fluorocarbon blood substitute or its main components in vitro. Neutrophil phagocytosis was determined by the method of chemiluminescence (CL) and the concentration of intracellular cAMP and cGMP were assessed. The results showed that the CL was inhibited while the level of cAMP was elevated. The alteration of cAMP seemed to be correlated with the inhibition of CL. Only did the emulsifier Poloxamer F-68 (F-68) of fluorocarbon blood substitute have the same effects. It is suggested that fluorocarbon blood substitute can inhibit neutrophil phagocytic function and the emulsifier F-68 may be responsible for it. The mechanism may be associated with the elevation of intracellular cAMP concentration. PMID- 2561937 TI - [Primary intestinal-type adenocarcinoma of the nasal cavity and paranasal sinuses]. AB - Forty-nine cases of adenocarcinoma of the nasal cavity and paranasal sinuses were reviewed, in which 5 cases showed a marked similarity to colonic adenocarcinoma. Diagnosis of intestinal-type adenocarcinoma chiefly depends on pathological examination and mucous staining. Although well differentiated, the intestinal type adenocarcinoma has a marked propensity for recurrence and a lethal potential. PMID- 2561938 TI - Transmembrane signalling in Saccharomyces cerevisiae. AB - Baker's yeast, a unicellular eukaryote, has been a model organism for biochemists, geneticists and most recently for molecular biologists. Pioneering biochemical studies were conducted on yeast, such as the study of glucose fermentation and amino acid metabolism. The powerful tools of yeast genetics have allowed a comprehensive study of important issues such as the cell cycle and meiosis. In recent years, it has been established that Saccharomyces cerevisiae, the most extensively characterized of the yeasts, shares key molecules and biochemical pathways with higher eukaryotes. For example, actin, tubulin, ubiquitin, calmodulin, GTP regulatory proteins, different protein kinases including protein tyrosine kinases, were all found to play central roles in yeast. Furthermore, structurally homologous proteins, as well as transcription regulating elements, of yeast and higher eukaryotes, including mammals, were shown to be structurally and functionally interchangeable. It has also been found that yeast can express human genes. Technically, yeasts are simple to handle, inexpensive to grow, complete a cell cycle within 90 min, and therefore can yield relatively quick results. These qualities are useful in biotechnological applications. Saccharomyces cerevisiae, can be genetically manipulated fairly easily, and has been tinkered with more than any other system. A cloned, in vitro mutated gene, can be transformed into wild type yeast and by homologous recombination, can replace the native gene and generate the desired mutant. Such manipulations, not possible yet in other eukaryotic cells, allow the precise definition of the role played by different genes and their domains. These unique features of Saccharomyces cerevisiae, together with rapidly evolving techniques of molecular biology, have made it a successful model organism for the study of numerous questions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561939 TI - Attenuation of neurotoxicity following anoxia or glutamate receptor activation in EGF- and hippocampal extract-treated neuronal cultures. AB - Neurotoxicity following anoxia or glutamate receptor activation was studied in primary neuronal cultures grown in serum-free, chemically defined CDM R12 medium. Exposure to 1 mM KCN, 0.5 mM kainic acid and 0.5 mM N-methyl-D-aspartate led to progressive neuronal degeneration. This damage was quantified by measuring lactate dehydrogenase released in the culture medium. The toxic effects were observed early during the development of the neuronal culture (from 4 days in vitro on) and seemed to be neuron-specific since astrocyte cultures were not affected. Chronic treatment of the neuronal cultures with epidermal growth factor at 10 ng/ml and hippocampal extract at dil. 1/833 (w/v) induced morphological alterations, increased beta-adrenergic receptor coupled adenylate cyclase activity, increased level of total lactate dehydrogenase activity in the case of epidermal growth factor-treated cultures, and attenuation of lactate dehydrogenase release following exposure to KCN or glutamate receptor agonists. The alterations observed are probably due to the proliferation and differentiation of glial cells in these treated cultures. This suggests that glial cells protect neurons in vitro from degeneration induced by anoxia or glutamate receptor activation. PMID- 2561940 TI - Multiple defects occur in the guanine nucleotide regulatory protein system in liver plasma membranes of obese (fa/fa) but not lean (Fa/Fa) Zucker rats: loss of functional Gi and abnormal Gs function. AB - Hepatocyte membranes from both lean and obese Zucker rats exhibited adenylate cyclase activity that could be stimulated by glucagon, forskolin, NaF and elevated concentrations of p[NH]ppG. In membranes from lean animals, functional Gi was detected by the ability of low concentrations of p[NH]ppG to inhibit forskolin-activated adenylate cyclase. This activity was abolished by treatment of hepatocytes with either pertussis toxin or the phorbol ester TPA, prior to making membranes for assay of adenylate cyclase activity. In hepatocyte membranes from obese animals no functional Gi activity was detected. Quantitative immunoblotting, using an antibody able to detect the alpha subunit of Gi, showed that hepatocyte plasma membranes from both lean and obese Zucker rats had similar amounts of Gi-alpha subunit. This was 6.2 pmol/mg plasma membrane for lean and 6.5 pmol/mg plasma membrane for obese animals. Using thiol pre-activated pertussis toxin and [32P]-NAD+, similar degrees of labelling of the 40 kDa alpha subunit of Gi were found using plasma membranes of both lean and obese Zucker rats. We suggest that liver plasma membranes from obese Zucker rats express an inactive Gi alpha subunit. Thus lesions in liver Gi functioning are seen in insulin-resistant obese rats and in alloxan- and streptozotocin-induced diabetic rats which also show resistance as regards the acute actions of insulin. Liver plasma membranes of obese animals also showed an impairment in the coupling of glucagon receptors to Gs-controlled adenylate cyclase, with the Kd values for activation by glucagon being 17.3 and 126 nM for lean and obese animals respectively. Membranes from obese animals also showed a reduced ability for high concentration of p[NH]ppG to activate adenylate cyclase. The use of [32P]-NAD+ and thiol-preactivated cholera toxin to label the 43 kDa and 52 kDa forms of the alpha-subunit of Gs showed that a reduced labelling occurred using liver plasma membranes from obese animals. It is suggested that abnormalities in the levels of expression of primarily the 52 kDa form of alpha-Gs may give rise to the abnormal coupling between glucagon receptors and adenylate cyclase in liver membranes from obese (fa/fa) Zucker rats. PMID- 2561941 TI - Unmasking a growth factor/oncogene-activated S6 phosphorylation cascade. PMID- 2561942 TI - Expression of a functional protein kinase C-gamma using a baculovirus vector: purification and characterisation of a single protein kinase C iso-enzyme. AB - The cloning of complementary DNAs for protein kinase C (PKC) has revealed a multi gene family of closely related protein kinases [Parker et al. (1986) Science 233, 853-859; Coussens et al. (1986) Science 233, 859-866]. In vivo, the distribution of the PKC isoenzymes follows a fairly tissue-specific pattern suggesting that functional differences exist between the members of this kinase family. To initiate a detailed characterisation of the individual isoenzymes, and as an alternative approach to purifying and separating the individual PKC types and their splice variants from mammalian tissues, we have expressed the bovine PKC type gamma in insect cells using a baculovirus expression vector. The bovine protein constitutes one of the major proteins in infected cells and can be purified to near homogeneity by a 2-step procedure. Analysis of the purified protein confirms that it has authentic mammalian PKC characteristics with respect to phospholipid dependence and phorbol ester binding. The bovine PKC gamma purified from infected cells is post-translationally modified and resolves into a doublet of molecular weights 82,000 and 84,000 upon SDS-polyacrylamide gel electrophoresis. These two size classes of polypeptides appear to result from differential phosphorylation as demonstrated by sensitivity to protein phosphatase treatment. The applicability and the potential of this system for the analysis of the various mammalian PKC isoenzymes is discussed. PMID- 2561943 TI - Metabolism of inositol 1- and 4-monophosphates in HL60 promyelocytic leukaemia cells. AB - The metabolism of inositol 1- and 4-monophosphates in HL60 promyelocytic leukaemia cells was studied. LiCl, BeCl2 and NaF inhibited the hydrolysis of both monophosphates with half maximal inhibition occurring at 1.2 mM, 0.3 microM, 0.25 mM (Ins 1P) and 0.14 mM, 0.56 microM, 0.28 mM (Ins 4P) respectively. Lithium was an uncompetitive inhibitor with respect to both substrates. Ins 4P inhibited the hydrolysis of Ins 1P in a concentration dependent manner, suggesting that it acts as a competing substrate for the same enzyme. Half maximal inhibition occurred at 120 microM Ins 4P. The lithium sensitive activity responsible for the metabolism of both monophosphates was present in a soluble fraction made from the cells. Taken together these data suggest that Ins 1P and Ins 4P are hydrolysed by a single soluble enzyme activity which is sensitive to inhibition by lithium, beryllium and fluoride. PMID- 2561944 TI - Purification of a serum factor which reverses dibutyryl cAMP induced differentiation. AB - We have previously shown [Grabham et al. (1988) Expt. Eye Res. 47, 123-133] that the adenovirus 12 transformed human retinoblast cell line (Ad 12 HER 10), like a number of other cell types of neuroepithelial origin, can be induced to differentiate in response to exposure to dibutyryl cAMP, and that this differentiation can be rapidly reversed by foetal calf serum. We present data here to show that a single protein, which we have termed differentiation reversal factor (DRF) and have isolated from serum, is responsible for this activity. Following reversal by DRF the growth rate of these cells was shown to be stimulated in serum-free medium. Using ammonium sulphate fractionation, gel filtration chromatography (Ultrogel AcA44), anion exchange chromatography (DEAE cellulose) and preparative gel electrophoresis, DRF has been purified to homogeneity, as judged by polyacrylamide gel electrophoresis in the presence and absence of SDS. The protein has a mol. wt of 72,000 and appears to exist in vivo as a monomer. The concentration of DRF in serum is in the range 100-500 micrograms/ml and is capable of reversing cAMP-induced differentiation of various primary human neuroepithelial cells at physiological concentrations. PMID- 2561945 TI - Inhibition of PI-kinase in rat liver membranes by F-. AB - In a number of membrane preparations GTP or its non-hydrolysable analogues stimulate the breakdown of PIP2 generating the second messengers, inositol triphosphate and diacylglycerol. The G-protein which couples the PIP2-specific phospholipase C with the receptors can also be activated by F-. However, the level of PIP2 is dependent upon the activity of a number of enzymes in the PI pathway. Besides stimulating the breakdown of PIP2, we report that in rat liver membranes F- also decreases the labelling of the polyphosphoinositides through inhibition of the PI-kinase. PMID- 2561946 TI - Effects of adrenalectomy on CRH regulation of ACTH release: adenylate cyclase activity, cyclic AMP-dependent protein kinase activity and ACTH release. AB - Corticotropin releasing hormone (CRH) stimulation of ACTH release and cyclic AMP mediated events involved in the control of ACTH release were compared in sham operated and adrenalectomized rats. CRH-stimulated adenylate cyclase activity was decreased in pituitary homogenates from adrenalectomized animals. CRH-stimulated cyclic AMP accumulation was essentially abolished and CRH-stimulated cyclic AMP dependent protein kinase (A-kinase) activity was decreased in freshly prepared anterior pituitary cells from adrenalectomized animals. Basal and CRH-stimulated ACTH release was elevated in these cells. Since ACTH release is increased in adrenalectomized rats despite the down regulation of CRH-linked pituitary mechanisms, we speculate that the site of action of disinhibition by corticosterone of ACTH release (or synthesis) following adrenalectomy is distal to the generation of cyclic AMP and/or that non-CRH mediated mechanisms assume a greater role in ACTH regulation following adrenalectomy. PMID- 2561949 TI - Rapid intracellular alkalinization of Saccharomyces cerevisiae MATa cells in response to alpha-factor requires the CDC25 gene product. AB - The alpha-factor mating pheromone induces a transient intracellular alkalinization of MATa cells within minutes after exposure to the pheromone, and is the earliest biochemical event that can be identified subsequent to the exposure. Dissipation of the pheromone induced pH gradient, using 2,4 dinitrophenol or sodium orthovanadate, does not inhibit the biological response of the yeast to the pheromone such as mating and 'schmoo' formation. These findings suggest that the pheromone mediated pH change per se is not a part of the transmembrane signalling but rather the consequence of a biochemical reaction triggered by the alpha-pheromone interaction with its receptor and may have a permissive effect on the pheromonal response. The cdc25ts mutation causes MATa cells to become nonresponsive to alpha-factor subsequent to a shift to the restrictive temperature, suggesting that the CDC25 gene product participates in the pheromone response pathway. PMID- 2561948 TI - Biphasic effect of sodium fluoride and guanyl nucleotides on binding to prostaglandin E2 receptors in rat epididymal adipocyte membranes. AB - Both NaCl and NaF promoted PGE2 binding to epididymal adipocyte membranes by apparent increase in the binding affinity. In order to distinguish between the effect of fluoride and the 'salt effect' of sodium on PGE2 binding, the effects of Mg2+ and guanyl nucleotides on PGE2 binding in the presence of NaCl or NaF were compared. Mg2+ decreased PGE2 binding; high NaF concentration abolished this inhibition, while increased NaCl concentrations did not affect the Mg2+ inhibition. In the presence of Mg2+ the effects of NaCl and NaF were additive. The enhancement of PGE2 binding by fluoride, unlike sodium, was dependent on the presence of Mg2+. Incubation of the membranes with GDP beta S, Gpp(NH)p, GTP or GTP gamma S increased PGE2 binding. Gradual increase in NaF concentrations in the presence of guanyl nucleotides resulted in stimulation of PGE2 binding at low NaF concentrations and inhibition of PGE2 binding at high NaF concentrations. No changes in the stimulatory action of NaCl on PGE2 binding were observed in the simultaneous presence of NaCl and guanyl nucleotides. A biphasic effect on PGE2 binding was observed with a wide concentration range of guanyl nucleotides. Treatment of the isolated membranes with cholera or pertussis toxins stimulated the adenylyl cyclase activity of the membranes, but failed to influence PGE2 binding. The implications of these findings are discussed. PMID- 2561947 TI - Regulation of epithelial sodium channel densities by vasopressin signalling. PMID- 2561950 TI - Role of cell calcium in alpha-1 adrenergic receptor control of arachidonic acid release from brown adipocytes. AB - Exposure of brown fat cells to phenylephrine, an agonist of alpha-1 adrenergic receptors, activates a phospholipase A2 which releases arachidonic acid. Since receptor activation of phospholipase A2 requires calcium, experiments were undertaken to define more precisely the role played by calcium in the regulation of enzyme activity. In this study, adipocytes were loaded with the fluorescent calcium chelator quin2 in order to buffer intracellular calcium and block receptor stimulated changes in its concentration. When quin2 loaded adipocytes were incubated in buffer containing 0.10 mM calcium, the ability of phenylephrine to stimulate release of arachidonic acid was severely reduced. At an intracellular quin2 concentration of 6.6 mM stimulated arachidonic acid release was inhibited by more than 50% and at 13 mM it was completely blocked. In contrast, phenylephrine stimulation of inositol phosphate accumulation was unaffected by quin2. Quin2 also did not affect the liberation of arachidonic acid in response to exogenous phospholipase C, A23187 or forskolin. The intracellular calcium antagonist TMB-8 also inhibited phenylephrine-stimulation of arachidonic acid release and this effect was reversed by ionomycin. Basal phospholipase A2 activity was increased by introduction of high calcium concentrations into cells rendered permeable with digitonin, but phenylephrine still caused a further increase in enzyme activity. These findings show a selective inhibition of phenylephrine activation of phospholipase A2 by either the chelation of intracellular calcium with quin2 or by the calcium antagonist TMB-8 and suggest an essential role for intracellular calcium in alpha adrenergic stimulation of enzyme activity. However, because phenylephrine still stimulates enzyme activity in cells rendered permeable with digitonin, we suggest that the action of phenylephrine cannot be attributed solely to changes in intracellular calcium. PMID- 2561951 TI - Activation of protein kinase C sensitizes the cyclic AMP signalling system of T51B rat liver cells. AB - Activation of protein kinase C (PKC) by phorbol esters (TPA) results in a modification of the cyclic AMP system leading to either attenuation or amplification of the cyclic AMP signal. In the non-neoplastic T51B rat liver cell line, TPA, when added to intact cells, had no effect on the basal level of cyclic AMP synthesis but caused a 1.5 fold amplification of the stimulation induced by beta-adrenergic agents, cholera toxin and forskolin. The effect appeared to be mediated by PKC since diacylglycerols caused the same amplification as did TPA while inactive phorbol esters were without effect. Phosphorylation of Gs or the catalytic subunit of adenylate cyclase by PKC is likely to be responsible for the enhancement of cyclic AMP synthesis. TPA also caused translocation of PKC; however, the time course of the translocation was longer than the time course of the enhancement of adenylate cyclase activity. Thus, the ability of TPA to amplify cyclic AMP synthesis is probably mediated by activation of PKC that is already present in the membrane. PMID- 2561952 TI - Spoilage of an acid food product by Clostridium perfringens, C. barati and C. butyricum. AB - Spoilage of canned pasteurized brined mung bean sprouts, acidified with citric acid to pH 4.0-4.5, was found to be caused by acid tolerant Clostridium spp. including the species barati, perfringens and butyricum. The pH limit for growth in the brine used were estimated 3.7, 3.7 and 4.0 respectively. Some of the isolated C. perfringens strains produced enterotoxins in sporulation media. The spores of the isolated anaerobes appeared to originate from mung beans, but C. barati and C. perfringens strains freshly isolated from dry beans, were unable to grow in acidified brine. During germination and sprouting of mung beans, the oxygen concentration decreased, while carbon dioxide concentration increased considerably, due to respiration of the sprouts and actively growing Enterobacteriaceae and lactobacilli. It was assumed that this allowed C. barati and C. perfringens strains to grow and acquire the observed unusual acid tolerance. After increasing aerobicity during sprouting, no growth of Clostridium spp. was observed, substantiating the assumption. PMID- 2561953 TI - Survival of foot-and-mouth disease virus in sausage meat products (Italian salami). AB - Determination of the survival of foot- and-mouth disease virus (FMDV) in fresh meat from experimentally infected swine and in several types of sausage meat (Italian salami) produced according to the technology widely applied by the principal Italian producers has been carried out. The purpose of the experiment was to assess if typical Italian salami can be considered safe with regard to the spread of FMD through international trade. The results obtained showed: (a) high titers of FMDV were detected in both muscle and fat tissues from animals slaughtered at the peak of the experimental disease; and (b) FMDV was not detectable in the above tissues 72 h after slaughtering and the same applies to the different types of salami tested 7 days after production. The above results were obtained in tissue cultures and confirmed through piglet inoculation. PMID- 2561954 TI - Antibacterial effect of the glucose oxidase-glucose system on food-poisoning organisms. AB - The antibacterial effect of the glucose oxidase-glucose system was studied on food-poisoning organisms including Staphylococcus aureus, Salmonella infantis, Clostridium perfringens, Bacillus cereus, Campylobacter jejuni, Listeria monocytogenes and Yersinia enterocolitica using automated turbidometry. The bacteria were grown in sterile-filtered meat medium which was either raw or heat denaturated. The results showed a clear growth inhibition with combinations of 0.5-1.0 mg/ml glucose and 0.5-1.0 IU/ml glucose oxidase. The growth inhibition was more effective in the heat-denaturated meat medium. The most resistant pathogens were Campylobacter jejuni and Listeria monocytogenes, however growth inhibition was still evident. The possible application of the glucose oxidase glucose system in food products inhibiting the growth of pathogens and spoilage organisms is discussed. PMID- 2561955 TI - Human glioblastoma cell lines have neuropeptide receptors for bombesin/gastrin releasing peptide. AB - Bombesin/gastrin-releasing peptide receptors were characterized in human glioblastoma cell lines. [125I]Gastrin-releasing peptide or ([125I]Tyr4)bombesin bound with high affinity to these cell lines. Binding to cell line U-118 was time dependent, reversible, and specific. ([125I]Tyr4)Bombesin bound with high affinity (Kd = 1.6 nM) to a single class of sites (Bmax = 30,000/cell). The C terminal of bombesin- or gastrin-releasing peptide was essential for high affinity binding. Bombesin- or gastrin-releasing peptide elevated the cytosolic Ca2+ levels in a dose-dependent manner. Because gastrin-releasing peptide, but not gastrin-releasing peptide, increased the cytosolic Ca2+ levels, the C terminal but not the N-terminal of GRP is essential for biological activity. These data indicate that biologically active bombesin receptors are present in human glioblastoma cell lines. PMID- 2561956 TI - Localization of cyclic GMP-dependent protein kinase in rat basal ganglia neurons. AB - Cyclic GMP-dependent protein kinase displays an uneven distribution in brain, being highly concentrated only in cerebellar Purkinje cells. Using DARPP-32 (dopamine- and cyclic AMP-regulated phosphoprotein, Mr 32,000) as exogenous substrate, and performing assays in the absence or presence of the protein inhibitor of cyclic AMP-dependent protein kinase, we have now identified both cyclic AMP-dependent and cyclic GMP-dependent protein kinase activities in the rat neostriatum and substantia nigra. Quinolinic acid-induced degeneration of neostriatal neurons and the straitonigral fibers emanating from neostriatal neurons decreased the activities of both cyclic nucleotide-dependent enzymes by 70-85% in the neostriatum, while cyclic GMP-dependent protein kinase was decreased by 44% and cyclic AMP-dependent protein kinase was decreased by 18% in the substantia nigra. In the basal ganglia, cyclic GMP-dependent protein kinase therefore appears enriched in striatonigral neurons, while cyclic AMP-dependent protein kinase is present both in striatonigral neurons and in other cells. The results indicate that cyclic GMP-regulated protein phosphorylation may play a role in the function of distinct basal ganglion neurons. PMID- 2561957 TI - Brain metabolites as 1H NMR markers of neuronal and glial disorders. AB - 1H NMR spectroscopy of human brain in vivo can be used to detect a number of cerebral metabolites including N-acetylaspartate, creatine + phosphocreatine and choline-containing compounds. We have used 1H NMR spectroscopy to analyse these signals in (i) biopsy material from both normal human brain and astrocytomas, and (ii) primary astrocyte cultures. On the basis of this analysis, we conclude that in vivo 1H NMR spectroscopy could play an important clinical role in the non invasive assessment of neuronal degeneration and proliferation of non-neuronal cells. PMID- 2561958 TI - The importance of the voxel size in clinical 1H spectroscopy of the human brain. AB - It is demonstrated that it is possible to acquire two volume selective 1H NMR spectra of human brain in vivo, consisting of voxels of 1.5 X 1.5 X 1.5 cm3, within 14 min with a good S/N ratio. This is mainly achieved by the application of a PRESS sequence generating a spin-echo of the VOI at 135 ms in conjunction with the STABLE technique by which two spectra can be recorded in an interlaced mode. The Bo homogeneity over such small voxels is considerably higher than over larger voxels. With these methodological improvements it is possible to observe morphological heterogeneity of tumors. The results indicate that spectral changes seem to correlate with the metabolic state of the tumor rather than the tumor type. Additionally the spectrum of a patient with multiple sclerosis suggests that even differentiation between tumors and other lesions might not be possible. PMID- 2561959 TI - Intraocular HIV-1-specific IgG synthesis in a patient with CMV retinitis. AB - A sight-threatening cytomegalovirus (CMV) retinitis is often associated with acquired immune deficiency syndrome (AIDS). We report the detection and quantitation of antibodies specific for CMV and human immunodeficiency virus type 1 (HIV-1) in the vitreous fluid and serum of an AIDS patient suffering from bilateral CMV retinitis. The ratio of the concentrations of HIV-1-specific immunoglobulin G (IgG) to total IgG in the vitreous fluid was found to be higher than that of the peripheral blood, indicating a local production of HIV-1 specific IgG synthesis within the ocular compartment. In contrast, CMV-specific IgG levels in serum were found to be higher than that of vitreous fluid, indicating an intact blood-ocular barrier. CMV and HIV were also isolated from the ocular tissues of this patient. These findings are consistent with the hypothesis that CMV retinitis may be associated with HIV-1 infection of the ocular tissues, which evokes an HIV-1-specific humoral immune response locally within the ocular compartment. PMID- 2561960 TI - Immunobiology of Langerhans cells on the ocular surface. II. Role of central corneal Langerhans cells in stromal keratitis following experimental HSV-1 infection in mice. AB - Stromal keratitis is typically the consequence of infection with herpes simplex virus type 1 (HSV-1). The pathogenesis of this disease remains elusive, although it is generally believed that there is an important immunological component. It has been proposed that stromal keratitis is mediated by virus-specific T lymphocytes of the delayed hypersensitivity type. However, while virtually all individuals infected with HSV-1 develop delayed hypersensitivity, only a small fraction actually develop stromal keratitis. To explain this discrepancy, we reasoned as follows: epidermal Langerhans cells are believed to be crucial to the induction of delayed hypersensitivity; since the cornea normally contains few or no cells of this type, the presence of Langerhans cells in the central corneal epithelium at the time of virus infection might promote the development of stromal keratitis. To test this hypothesis, cautery wounds of central regions of mouse corneas were used to induce migration of Langerhans cells into the corneal epithelium. These mice were then infected with HSV-1 on the ipsilateral snout, an infection that results in zosteriform spread of virus via the trigeminal nerve into the anterior segment of the ipsilateral eye within 3-5 days after inoculation. We found that the eyes of cauterized mice displayed a very high incidence of severe stromal keratitis. By contrast, non-cauterized corneas of control, snout-infected mice displayed much less evidence of stromal disease. Moreover, the rapidity of onset of systemic delayed hypersensitivity to HSV-1 was accelerated in the mice with cauterized corneas, compared to the controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561961 TI - Cholera toxin as a mucosal adjuvant for respiratory antibody responses in mice. AB - Cholera toxin was investigated as an adjuvant for anti-virus antibody responses in the respiratory mucosa of mice. Two methods of applying cholera toxin were evaluated: oral administration and intranasal administration. Oral immunization with Sendai virus in the presence of cholera toxin effectively primed for respiratory anti-viral antibody responses, whereas oral immunization with Sendai virus alone was ineffective in this respect. In nasal washes, IgA was the predominant anti-viral antibody enhanced by oral cholera toxin; in bronchoalveolar washes, the enhanced anti-viral antibodies included IgG, IgA, and IgM. Effects of direct administration of cholera toxin to the respiratory mucosa on respiratory anti-viral antibody responses depended on the method of anesthesia used during immunization. With inhalation anesthesia (ether), cholera toxin had no adjuvant effect on respiratory antibody responses to coadministered Sendai virus. In contrast, under parenteral anesthesia (i.e., intraperitoneal ketamine), mice which received cholera toxin and Sendai virus via the respiratory tract showed significantly higher anti-viral IgA and IgG antibody titers in nasal washes and IgG antibody in bronchoalveolar washes than mice which received the virus only. PMID- 2561962 TI - [Effects of VIP (vasoactive intestinal peptide) on plasma membrane ATPases from rat enterocytes]. AB - We studied the effect of VIP on the ATPase activities from basolateral membranes prepared from rat enterocytes. Under the standard conditions of assay for membrane ATPases (millimolar ATP concentration) VIP has no effect, neither on the Na, K ATPase activity (ouabain sensitive) nor on the Mg ATPase activity (ouabain insensitive). These results suggest that short-term effects of VIP on ionic permeability and metabolism of enterocytes, are not mediated through modifications of the Na/K ratio by the Na, K ATPase or through modifications of another membrane ATPase activities. PMID- 2561963 TI - [Effects of non-lethal scalding on the density and affinity of beta adrenoreceptors in cardiac muscle in rats]. AB - Our results indicate changes in both Bmax and KD of the beta-adrenergic receptors in the ventricular heart muscle of the rat, submitted to non-lethal scalding, even in the early posttraumatic period. The changes of heart, plasma and urine noradrenaline and adrenaline contents indicate significant increase in sympatho adrenal activity after non-lethal scalding. The parallel changes of the beta adrenergic receptor characteristics, which depend on the changes of catecholamines in the heart muscle and plasma only at certain time intervals after trauma, tend to preserve normal-control values. PMID- 2561964 TI - Effect of divalent cations on the structure of dipalmitoylphosphatidylcholine and phosphatidylcholine/phosphatidylglycerol bilayers: an 2H-NMR study. AB - The interactions of CaCl2 or MgCl2 with multilamellar phospholipid bilayers were studied by 2H-NMR. Two model membrane systems were used: (1) dipalmitoylphosphatidylcholine (DPPC) bilayers and (2) bilayers composed of a mixture of phosphatidylcholine and phosphatidylglycerol at a molar ratio of 5:1. Addition of 0.25 M CaCl2 to DPPC bilayers resulted in significant uniform increase of the order parameters of the lipid side chains; the effect of 0.25 M MgCl2 was insignificant. Both phosphatidylcholine and phosphatidylglycerol components of the mixed bilayers were affected by the presence of 0.25 M CaCl2 and, to a much smaller degree, by MgCl2. The addition of Ca2+ induced significantly larger increase of the order parameters of the phosphatidylcholine component. The results are consistent with the long-range effects of Ca2+ binding on the packing of the lipid membranes. PMID- 2561965 TI - [Comparative study of Uromiron and 38% Lipiodol in sialography]. AB - A comparative study was undertaken to assess the properties of uromiron and lipiodol 38%, which are used as radiopaque materials in sialographic techniques, on 25 young adult outpatients of both sexes that were seen at the Dental School, University of Chile, Santiago. It was found that, although the radiopaque properties of uromiron were less than lipiodol 38%, the can be improved by increasing the amount of material injected into the duct system of the salivary glands. Likewise, uromiron exhibited a proper biological tolerance and a rapid emptying time, therefore the use of uromiron in sialographic techniques seems to be a proper alternative to lipiodol 38%. PMID- 2561966 TI - [Neuroectodermal pigmented tumor in an infant]. PMID- 2561967 TI - Kassinin-like neuropeptides in the control of body fluid homeostasis. PMID- 2561968 TI - Protein kinase C is involved in laminin stimulation of neurite outgrowth. AB - We are investigating the intracellular events involved in the induction of neurite outgrowth. The phorbol ester TPA, an activator of protein kinase C, potentiates neurite outgrowth from ciliary ganglion neurons cultured on suboptimal laminin concentrations, but not on optimal laminin concentrations. TPA also stimulates growth on fibronectin and collagen similar to that observed on laminin under control conditions. Manipulations that elevate intracellular cAMP levels (expected to activate A kinase) reduce neurite outgrowth on laminin. The protein kinase C inhibitors H7 and sphingosine inhibit neurite outgrowth on laminin in a reversible and dose-dependent manner. H7 does not inhibit the process outgrowth induced by concanavalin A in the same neurons. The results suggest that activation of protein kinase C is an important step in the neurite outgrowth caused by laminin binding to its receptor(s). PMID- 2561969 TI - Bicarbonate dependence of glutamate receptor activation by beta-N-methylamino-L alanine: channel recording and study with related compounds. AB - beta-N-methylamino-L-alanine (BMAA) is a neurotoxic glutamate agonist possibly responsible for the neuronal degeneration found in the Guam amyotrophic lateral sclerosis-Parkinsonism-dementia complex. The basis for glutamate receptor activation by BMAA has been unclear, as BMAA lacks the omega electronegative moiety characteristic of other excitatory amino acids. We recently reported that the neuroexcitatory and neurotoxic effects of BMAA depend strongly on the presence of bicarbonate ions and proposed that an interaction between bicarbonate and the beta amino group of BMAA produces a molecular configuration appropriate for activating glutamate receptors. We now report that bicarbonate potentiates the ability of BMAA to open NMDA receptor-activated channels in isolated membrane patches. Furthermore, the neurotoxic and neuroexcitatory effects of two structural analogs of BMAA, DL-2,4-diaminobutyrate and DL-2,3-diaminopropionate, were also potentiated by bicarbonate. These findings support the bicarbonate cofactor hypothesis for BMAA action and provide direct evidence that it may be generalizable to certain other compounds. PMID- 2561970 TI - Two novel GABAA receptor subunits exist in distinct neuronal subpopulations. AB - Two cDNAs encoding novel GABAA receptor subunits were isolated from a rat brain library. These subunits, gamma 2 and delta, share approximately 35% sequence identity with alpha and beta subunits and form functional GABA-gated chloride channels when expressed alone in vitro. The gamma 2 subunit is the rat homolog of the human gamma 2 subunit recently shown to be important for benzodiazepine pharmacology. Cellular localization of the mRNAs encoding the gamma 2 and delta subunits in rat brain revealed that largely distinct neuronal subpopulations express the two subunits. The delta subunit distribution resembles that of the high affinity GABAA receptor labeled with [3H]muscimol; the gamma 2 subunit distribution resembles that of GABAA/benzodiazepine receptors labeled with [3H]flunitrazepam. These findings have implications for the composition of two different GABAA receptor subtypes and for information processing in networks using GABA for signaling. PMID- 2561971 TI - Primary cultures of mouse spinal cord express the neonatal isoform of the inhibitory glycine receptor. AB - Expression of the inhibitory glycine receptor complex was investigated in primary cultures of fetal mouse spinal cord using sensitive immunomethods. In these cells, glycine receptor is predominantly of the neonatal isoform characterized by a low affinity for the antagonist strychnine. It contains a ligand binding subunit that differs from that of the adult receptor in antigenic epitopes and apparent molecular weight. Whereas in vivo the neonatal receptor isoform is completely replaced by the adult isoform within 3 weeks after birth, this exchange of subtypes is not seen in culture. The increased expression of the cytoplasmic glycine receptor-associated polypeptide of 93 kd occurring after birth is also seen under culture conditions. Purification of glycine receptor from cultures yielded polypeptides of 49 kd and 93 kd, suggesting that the membrane-spanning core of the neonatal receptor may be a homooligomer composed of 49 kd subunits. About half of the 49 kd subunit is cleaved by trypsinization of the cultures, indicating a predominant cell surface localization of the receptor. Pulse-labeling experiments revealed the 49 kd subunit to be a metabolically stable glycoprotein (half-life approximately 2 days). After its synthesis, a transition time of 30-45 min is required for acquisition of a strychnine binding conformation. PMID- 2561972 TI - Developmental changes in the cellular composition of a brain nucleus involved with song learning in zebra finches. AB - Using a double-labeling technique to characterize projection neurons and androgen target cells, we examined ontogenetic changes in the cellular composition of IMAN, a forebrain nucleus that plays an important role in song learning during a restricted period of male zebra finch development. This nucleus undergoes a massive loss of neurons during the time of song acquisition. We report that during the period of cell loss in IMAN, neither the property of projecting to an efferent target nor the ability to concentrate androgens is able to spare neurons from ontogenetic cell death. Furthermore, we report that, at the time when IMAN ceases to influence song production, a large proportion of androgen-sensitive cells that do not make an efferent projection lose the ability to accumulate androgens. PMID- 2561973 TI - The RII subunit of cAMP-dependent protein kinase binds to a common amino-terminal domain in microtubule-associated proteins 2A, 2B, and 2C. AB - Three products of the MAP2 gene are known: MAP2A and MAP2B (Mr approximately 200,000) and MAP2C (Mr 70,000). The structural relationship between these MAPs and the basis for their diversity in size are unknown. Previously, we found that a significant fraction of type II cAMP-dependent protein kinase was associated via its regulatory subunits with MAP2A and MAP2B. We now use an antibody prepared against the microtubule binding domain of MAP2A and MAP2B to identify MAP2C. All three forms of MAP2 bound to cAMP affinity columns and reacted with 32P-labeled RII in a blot overlay assay. By assaying proteolytic fragments of MAP2A and MAP2B as well as segments of MAP2 expressed in E. coli, the binding site for RII was localized to an 83 amino acid stretch at the distal (amino-terminal) end of the MAP2 arm domain. Therefore, the microtubule binding and RII binding domains are located at extreme opposite ends of MAP2A and MAP2B, and both are conserved in the much shorter MAP2C. PMID- 2561974 TI - Chromosomal localization of GABAA receptor subunit genes: relationship to human genetic disease. AB - Hybridization of GABAA receptor probes to human chromosomes in situ and to DNA from sorted human chromosomes has localized the genes encoding a beta subunit and three isoforms of the alpha subunit. The alpha 2 and beta genes are both located on chromosome 4 in bands p12-p13 and may be adjacent. The alpha 1 gene is on chromosome 5 (bands q34-q35) and the alpha 3 gene is on the X chromosome. The alpha 3 locus was mapped also on the mouse X chromosome using genetic break-point analysis in an interspecies pedigree. The combined results locate the human alpha 3 gene within band Xq28, in a location that makes it a candidate gene for the X linked form of manic depression. PMID- 2561975 TI - Developmental regulation of nerve growth factor and its receptor in the rat caudate-putamen. AB - In prior studies, nerve growth factor (NGF) administration induced a robust, selective increase in the neurochemical differentiation of caudate-putamen cholinergic neurons. In this study, expression of NGF and its receptor was examined to determine whether endogenous NGF might serve as a neurotrophic factor for these neurons. The temporal pattern of NGF gene expression and the levels of NGF mRNA and protein were distinct from those found in other brain regions. NGF and high-affinity NGF binding were present during cholinergic neurochemical differentiation and persisted into adult-hood. An increase in NGF binding during the third postnatal week was correlated with increasing choline acetyltransferase activity. The data are consistent with a role for endogenous NGF in the development and, possibly, the maintenance of caudate-putamen cholinergic neurons. PMID- 2561976 TI - Differential subcellular localization of the RI and RII Na+ channel subtypes in central neurons. AB - Immunocytochemical localization of Na+ channel subtypes RI and RII showed that RI immunoreactivity is relatively low and homogeneous along the rostral-caudal extent of sagittal brain sections, whereas RII staining is heterogeneous and relatively dense in the forebrain, substantia nigra, hippocampus, and cerebellum. The somata of the dentate granule cells, hippocampal pyramidal cells, cerebellar Purkinje cells, and spinal motor neurons are immunoreactive for RI but not RII. In contrast, areas rich in unmyelinated nerve fibers, such as the mossy fibers of the dentate granule cells, the stratum radiatum and stratum oriens of the hippocampus, and the molecular layer of the cerebellum, are strongly immunoreactive for RII but not RI. Differential regulation of expression of RI and RII genes may allow differential modulation of Na+ channel density in somata and axons. The sites of RI localization correlate closely with sites where sustained Na+ currents have been recorded. PMID- 2561977 TI - A novel alpha subunit in rat brain GABAA receptors. AB - Two cDNAs (alpha 1 and alpha 4) from rat brain cDNA libraries encode isoforms of the alpha subunit of the GABA/benzodiazepine receptor, which differ at 30% of their amino acid residues. Northern blot analysis and in situ hybridization histochemistry show that alpha 1 and alpha 4 mRNAs have distinct sizes and distinct regional and cellular distributions in rat brain: both mRNAs are found in the cortex and hippocampus; however, only the alpha 1 mRNA is detected in the cerebellum. We injected RNA transcribed from alpha 1 and alpha 4 cDNAs into Xenopus oocytes, together with an RNA for a rat beta subunit. We obtained GABA dependent inward currents that were reversibly blocked by picrotoxin. Picrotoxin alone, applied to oocytes producing the alpha and beta polypeptides, elicited an outward current. We suggest that these polypeptides together produce GABA-gated ion channels that can also open spontaneously. PMID- 2561978 TI - SCIP: a glial POU domain gene regulated by cyclic AMP. AB - We have isolated cDNA clones encoding SCIP, a POU domain gene expressed by myelin forming glial of the central and peripheral nervous systems. In purified Schwann cells cultured in the absence of neurons, expression of SCIP is suppressed. This suppression is relieved by cAMP, and induction of SCIP mRNA by this second messenger precedes cAMP induction of myelin-specific genes. Similarly, SCIP expression in vivo precedes full expression of myelin-specific genes in developing oligodendrocytes and Schwann cells. The sequence of the SCIP POU domain is identical to that of Tst-1, a recently identified member of a family of POU domain genes expressed by restricted subsets of neurons. Our results demonstrate that SCIP is also expressed by myelin-forming glia and suggest that it plays a central role in the progressive determination of these cells and their commitment to myelination. PMID- 2561979 TI - The in vivo and in vitro effects of interleukin-1 and tumor necrosis factor on murine cytomegalovirus infection. AB - The effects of tumor necrosis factor (TNF) and interleukin-1 (IL-1) on infection with murine cytomegalovirus (MCMV) were investigated in vitro and in vivo. The addition of each of these cytokines (at 1 ng/ml) to tissue culture monolayers 24 hr prior to MCMV challenge produced a reproducible decrease in vital titer (from 1 x 10(8) pfu to approximately 4 x 10(6) pfu for both cytokines). There was no further increase in this effect when a 10 or 100 ng/ml of each of these cytokines was employed. Despite these in vitro effects, the pretreatment of suckling, weanling, or adult mice with 80 or 400 ng of TNF or IL-1 alone, or 80 ng of each cytokine together, had no effect on the survival of mice following MCMV. Similarly, neither of these cytokines adversely influenced the protective effects of hyperimmune anti-MCMV antiserum; that is, they did not attenuate the protection conferred by the antiserum nor affect the protective effects of subtherapeutic doses of the antiserum. We conclude that despite promising antiviral effects against MCMV in vitro, these agents do not result in a useful therapeutic effect in vivo. Moreover, despite the ability of IL-1 to induce ACTH and corticosterone in mice, IL-1 treatment did not increase the mortality to CMV. PMID- 2561980 TI - Inositol phosphate metabolism: further problems and some solutions. PMID- 2561981 TI - Cross-talk between different receptor-effector systems in the mammalian CNS. PMID- 2561982 TI - Insulin mediators revisited. PMID- 2561983 TI - Phorbol ester inhibits polyphosphoinositide phosphodiesterase activity stimulated by either Ca2+, fluoride or GTP analogue in HL60 membranes and in permeabilized HL60 cells. AB - The effect of PMA (phorbol 12-myristate, 13-acetate) on PPI-pde (polyphosphoinositide phosphodiesterase) activity in the promyelocytic cell-line HL60 was examined. HL60 cells were pretreated with PMA in a time- and concentration-dependent manner and PPI-pde activity was monitored both in streptolysin O-permeabilized cells and in membranes. PPI-pde activity was stimulated by either GTP gamma S (guanosine 5'-[gamma-thio]triphosphate), fluoride or Ca2+. Both the Ca2(+)-stimulated and the G protein-mediated PPI-pde activity in permeabilized HL60 cells is maximally inhibited (70-90%) after 60 min pretreatment of intact cells with 10nM PMA. PPI-pde activity can also be observed in membranes prepared from HL60 cells although this activity represents only 10% of the total activity seen in permeabilized cells. In membranes, where PPI-pde activity can also be stimulated by either via the G-protein or directly by Ca2+, PMA pretreatment was also inhibitory regardless of the mode of activation. We suggest that both the membrane-bound PPI-pde activity and that present in the permeabilized cells are targets for protein phosphorylation by protein kinase C leading to inhibition of the catalytic function. PMID- 2561984 TI - The beta-adrenoceptor is precoupled to Gs in chicken erythrocyte membranes. AB - In this study we seek to elucidate the mechanism of hormone-independent adenylate cyclase stimulation by Gpp(NH)p in chicken erythrocyte membranes, and the inhibition of this stimulation by propranolol. Membrane treatment with isoprenaline + GMP increased Gpp(NH)p stimulation to near maximal levels [obtainable with isoprenaline + Gpp(NH)p], but reduced stimulation by NaF. The stimulation by Gpp(NH)p was stereoselectively inhibited by propranolol, but not by equal concentrations of the local anaesthetic lignocaine. Propranolol's inhibitory action was abolished following membrane treatment with isoprenaline/GMP. In contrast to its inhibition of Gpp(NH)p stimulation, propranolol did not alter Gpp(NH)p-mediated 3H-GDP release from membranes. The polyene antibiotic filipin, which uncouples receptor (R) from Gs, also abolished Gpp(NH)p stimulation and this effect was partly overcome by membrane treatment. These results are consistent with a model in which free R exists in equilibrium with precoupled R.Gs complexes in the absence of hormone. These complexes are activated by Gpp(NH)p and dissociated by antagonists. The existence of such complexes is a prerequisite for Gpp(NH)p stimulatory action. PMID- 2561985 TI - Benzylisoquinoline compounds inhibit the ability of calmodulin to activate cyclic nucleotide phosphodiesterase. AB - Benzylisoquinoline compounds antagonised the ability of calmodulin (CaM) to stimulate the activity of calmodulin-dependent cyclic nucleotide phosphodiesterase (CaM-PDE). This 'anti-CaM' activity was related to the hydrophobicity of the non-polar terminal region of the antagonist molecule. Antagonistic potency increased with the increase of hydrophobicity; the anti-CaM activity did not change when the polar terminus was a tertiary amine or quarternary amine. The anti-CaM potency was greater for bisbenzylisoquinoline compounds than for monobenzylisoquinoline compounds. Among the bisbenzylisoquinoline compounds anti-CaM pathway was: D3 greater than D2 berbamine greater than daurisoline greater than dauricine. Compound D3, which exhibited an IC50 value of 2.8 microM, was one of the most potent calmodulin antagonists, among benzylisoquinoline compounds, so far reported. PMID- 2561986 TI - Potassium as a signal for both proliferation and differentiation of rabbit retinal (Muller) glia growing in cell culture. AB - Retinal glial (Muller) cells were grown from explants of early postnatal rabbit retinae. The resulting monolayers of flat cells were exposed to control media (containing 5.85 mM K+), and to media with enhanced K+ concentrations (10 and 20 mM) or arginine-vasopressin (AVP, 20 micrograms/ml) or epithelial growth factor (EGF, 10 ng/ml). Autoradiographically, protein synthesis was quantified as L-[3H] lysine incorporation, and DNA synthesis as [3H]-thymidine incorporation. Furthermore, the activity of Na+,K(+)-ATPase was measured radiochemically. Short exposure to either moderately enhanced K+ concentrations (10 mM) or to AVP, stimulated L-[3H]-lysine incorporation into the cells. Long-lasting exposure to either high K+ concentrations (20 mM) or to EGF stimulated [3H]-uptake. The Na+,K(+)-ATPase activity of cell cultures increased with increasing K+ concentration of the media. It is suggested that release of K+ by active neuronal compartments stimulates local protein synthesis of glial cells, resulting in the formation of glial sheaths with active K+ uptake capacity. Strong K+ release may even induce glial proliferation. PMID- 2561987 TI - Regulation of protein kinases by pseudosubstrate prototopes. PMID- 2561988 TI - Bradykinin-induced changes in phosphoinositides, inositol phosphate production and intracellular free calcium in cultured bovine aortic endothelial cells. AB - Acute hydrolysis of phosphoinositides has been demonstrated in bovine aortic endothelial cells (BAEC) treated with bradykinin (BK) (10(-7)M). The first phosphoinositide to decrease was phosphatidylinositol-4,5-bisphosphate (PIP2) indicating this to be the initial substrate of phospholipase action. Other lipid changes associated with the stimulation of BAEC were an increase in diacylglycerol (DAG) and arachidonic acid (AA) with a sustained production of phosphatidic acid (PA). The changes in cell phospholipids were accompanied by the release of inositol phosphates. Inositol-1,4,5-trisphosphate (Ins-1,4,5-P3) was produced within 10 s of stimulation with BK. There was no evidence for the production of inositol-1,3,4-trisphosphate. The release of ionic calcium (Ca2+) intracellularly was demonstrated. The timecourse of the rise in intracellular Ca2+ was consistent with the timecourse of production of IP3. Intracellular Ca2+ rose from 127 +/- 21 nM to 462 +/- 27 nM. The Ca2+ peak was at 7.0 +/- 0.4 s and took 3 min to reach a steady state which remained above the basal level. When extracellular Ca2+ was depleted in the extracellular medium a spike of intracellular Ca2+ release was measured with an immediate return to basal. Entry of extracellular Ca2+ into the cell after ionophore A23187 treatment does not induce inositol phosphate release, indicating that phosphoinositide hydrolysis is likely to be the cause rather than consequence of the elevation in cytosolic Ca2+. These data indicate action of phospholipase C (PLC) on PIP2 after BK stimulation of BAEC with the subsequent production of InsP3 causing the resulting intracellular Ca2+ release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2561989 TI - Lipid signalling pathways in normal and ras-transfected NIH/3T3 cells. AB - The role of ras oncogenes in cellular signalling pathways involving phospholipid breakdown was studied in untransfected and proto-H-ras and mutated H-, K- and N ras transfected NIH/3T3 cells. When the cells were grown at low cell densities, all of the ras transfected cells had 2-4 fold higher diacylglycerol (DAG) levels compared to growing NIH/3T3 cells. At high cell densities, DAG levels decreased in the former and increased in contact inhibited NIH/3T3 cells. In this regard, only cells transformed by mutated cellular and viral H-ras oncogenes (but not by the H-ras proto-oncogene) had elevated DAG levels compared to contact inhibited NIH/3T3 cells. The basal levels of inositol phosphates in ras transfected cells were not significantly different from NIH/3T3 cells and did not vary with cell density. Thus, the elevated DAG levels are not a consequence of increased phosphoinositide hydrolysis. The latter was stimulated by serum and bombesin only in normal and proto-H-ras transfected cells. In contrast, stimulation by bradykinin was observed only in cells transformed by mutated cellular ras oncogenes. Furthermore, aluminum fluoride stimulated phosphoinositide breakdown in the latter cells indicating that there was no uncoupling of the G protein from phospholipase C. Treatment of ras transfected cells with dibutyryl cyclic AMP (DB cAMP), which causes an inhibition of growth and a reversal of the transformed morphology, did not alter the basal levels of inositol phosphates, DB-cAMP, however, did lower DAG levels in some of the transformed cell lines, but elevated DAG levels in low density NIH/3T3 cells. These findings indicate that the ras gene product p21 is not involved in phosphoinositide hydrolysis and that DAG levels do not correlate with cell growth in either normal or ras transfected NIH/3T3 cells. Thus, p21 appears to alter cell growth through mechanism(s) independent of lipid signalling pathways. PMID- 2561990 TI - Pharmacologic implications of alpha-adrenoreceptor interactive parameters for epinephrine enantiomers in the rat vas deferens. AB - After alkylation of a fraction of the total alpha-adrenoreceptors by phenoxybenzamine in rat vas deferens, the dissociation constants of (-)- and (+) epinephrine in functional studies were 7 X 10(-7) M and 2 X 10(-5) M, respectively. In the adrenoreceptor-containing tissue fraction, when 3H-labeled WB4101 was used as the interacting ligand, for each enantiomer two affinity sites were found. Only the low-affinity dissociation constant for each isomer correlates with the constant obtained from the functional studies. If the change in Gibb's free energy, delta G degrees, is calculated from the low-affinity binding constants, the values -8.1 and -6.2 kcal/mol for (-)- and (+)-isomer, respectively, are obtained. The small difference in the value between isomers is consistent with the view that the benzylic hydroxyl group of the (-)-isomer forms a hydrogen bond with the receptor. The interaction of epinephrine with this receptor appears to be driven largely by the entropy of the drug-receptor interaction with only a small nonstereoselective contribution from the enthalpy of interaction. PMID- 2561991 TI - Stereochemical influences upon the opioid ligand activities of 4-alkyl-4 arylpiperidine derivatives. AB - The synthesis and stereochemistry (configuration and preferred solute conformation) of some 4-alkyl (methyl, n-propyl, isobutyl)-4-(3-hydroxy-phenyl)-1 methylpiperidines and corresponding 3-methyl diastereoisomeric pairs are reported, together with their in vivo and in vitro activities as opioid ligands. All potent agonists exhibit a preference for axial 4-aryl chair conformations when protonated, and stereochemical analogies with rigid opioids of the benzomorphan class are discussed. Antagonist properties are found in compounds with preference for equatorial 4-aryl chairs, notably the cis 3,4-dimethyl derivative. PMID- 2561992 TI - Insulin and IGF-I stimulate phosphorylation of their respective receptors in intact neuronal and glial cells in primary culture. AB - Previous studies have shown that insulin and IGF-I bind to their respective receptors and stimulate autophosphorylation of the receptor beta subunits in detergent extracts of neuronal and glial cells. In the present study, intact neuronal and glial cells in primary culture have been utilized to characterize insulin- and IGF-I-stimulated phosphorylation of their receptors. Following [32P]orthophosphate labelling and stimulation by insulin or IGF-I, the cells were solubilized and the phosphorylated receptors were partially purified on wheat germ agglutinin--agarose columns, and immunoprecipitated using anti phosphotyrosine or anti-insulin receptor antibodies. Insulin stimulated the phosphorylation of its receptor beta subunit (95 kD phosphoprotein) in a dose dependent manner, within at least 20 seconds in both neuronal and glial cells. Additionally, a 102-kD phosphoprotein was observed in insulin-stimulated neuronal cells. Maximal stimulation of receptor phosphorylation occurred at 1 minute for the glial cells, and 10 minutes for the neuronal cells. IGF-I stimulated the phosphorylation of two phosphoproteins in intact neuronal and glial cells; a 95 kD protein and a 102-kD protein, in a dose-dependent manner. These observations demonstrate that both insulin and IGF-I stimulate the phosphorylation of the beta subunits of their respective receptors in brain cells in a similar fashion to their effects on receptors from nonneural tissues. PMID- 2561994 TI - Inositol 1,4,5-trisphosphate binding sites in the brain: regional distribution, characterization, and alterations in brains of patients with Parkinson's disease. AB - [3H]Inositol 1,4,5-trisphosphate [( 3H]Ins-(1,4,5)P3) binding studies were done on the human brain obtained at autopsy. The specific [3H]Ins(1,3,4,5)P3 binding sites in the cerebral and cerebellar cortices consisted of a single component with a high affinity (Kd = 11.3 and 16.5 nM, Bmax = 0.8 and 6.4 pmol/mg protein, respectively). The binding of [3H]Ins(1,4,5)P3 was potently inhibited by Ins(1,4,5)P3, in a nanomolar concentration, while other inositol phosphates and inositol were either much less potent or did not inhibit binding. The binding sites for [3H]Ins(1,4,5)P3 were discretely localized and were in the order: cerebellum much greater than basal ganglia, cerebral cortex greater than rhinencephalon greater than diencephalon, mesencephalon. There was an age-related loss of [3H]Ins(1,4,5)P3 binding in the frontal cortex. In the brains of patients with Parkinson's disease, [3H]Ins(1,4,5)P3 binding sites were reduced by about 50% in the caudate nucleus, putamen, and pallidum, while there were no differences in the frontal cortex, as compared to findings in the age-matched controls. Our findings suggest that [3H]Ins(1,4,5)P3 binding sites are closely linked to neural elements in the human brain. PMID- 2561995 TI - Epstein-Barr virus-associated hemophagocytic syndrome: clinical presentation and treatment. PMID- 2561993 TI - Localization of striatal opioid gene expression, and its modulation by the mesostriatal dopamine pathway: an in situ hybridization study. AB - In situ hybridization was used to study the macroscopic distribution and regulatory control of proenkephalin mRNA and prodynorphin mRNA in rat striatum. While proenkephalin mRNA was widely distributed throughout the striatum, levels of prodynorphin mRNA were highest in the medial and ventral portions of the striatum. Furthermore, in contrast to the results for proenkephalin mRNA, the levels of prodynorphin mRNA appeared higher in the nucleus accumbens than in the striatum. The mesostriatal dopaminergic pathway was destroyed by discrete, unilateral injection of 6-hydroxy-dopamine (6-OHDA) into either the substantia nigra or the neighboring ventral tegmental area (VTA). Lesions of the substantia nigra caused a dramatic ipsilateral increase in the hybridization signal for proenkephalin mRNA, but no change was observed in the hybridization signal for prodynorphin mRNA. Similar effects were seen with VTA lesions. Since destruction of the mesostriatal dopamine system elevates the levels of proenkephalin mRNA, but not of prodynorphin mRNA, in the striatal target neurons, it appears that the mesostriatal pathway exerts a tonic and selective suppression of striatal proenkephalin gene expression at the mRNA level. PMID- 2561997 TI - CNS cryptococcosis: unusual aspects. AB - Eleven patients with CNS cryptococcal infection are reviewed. The most prominent symptom was headache, present in all patients. The clinical manifestations were the direct result of the meningitis itself or a consequence of intracranial cryptococcal granulomata or hydrocephalus, these latter 2 complications being demonstrable on CT head scan. In the 2 patients who also had MRI scans, additional parenchymal lesions were revealed which had not been detected by CT. Combined amphotericin B and 5-fluorocytosine therapy was the treatment of choice, but in 3 patients fluconazole was also used. Chronic oral therapy with this agent has maintained a good clinical response in one patient who failed to respond to traditional antifungal therapy. PMID- 2561996 TI - Long-term follow-up after unilateral nephrectomy and radiotherapy for Wilms' tumour. AB - Twenty-nine patients who had had unilateral nephrectomy for Wilms' tumour in one hospital were known to have survived more than 12 years. Sixteen agreed to attend for clinical review, of whom 14 had estimation of serum creatinine, 24-h urine protein excretion and endogenous creatinine clearance. The follow-up period was 13-26 years (median 17 years). All but one had had radiotherapy and all had chemotherapy (actinomycin D, 16; vincristine, 5). Some degree of kyphoscoliosis was present in all except the patient who did not receive radiotherapy. Four patients had diastolic blood pressure 90 mmHg or greater. Two patients had mild proteinuria (392, 361 mg/day). Serum creatinine ranged from 53 to 125 mumol/l and endogenous creatinine clearance ranged from 39 to 173 ml/min (median 81, mean 89). Of the 7 patients who were 20-26 years post-nephrectomy, 2 were hypertensive and 1 had elevated urinary protein excretion. We conclude that the long-term prognosis of unilateral nephrectomy in childhood is good. PMID- 2561998 TI - A kinetic analysis of lymphocyte 5'-nucleotidase in B-cell clones from control subjects and patients with primary hypogammaglobulinaemia. AB - The affinity of lymphocyte 5'-nucleotidase (5NT) for its substrate adenosine monophosphate (AMP) was studied in Epstein-Barr virus immortalized B-lymphocyte clones from healthy controls and patients with primary hypogammaglobulinaemia. Km values for AMP for 5NT in most of the patient clones were found to be significantly increased compared to B-cell clones from normal subjects, whereas Vmax values were within the normal range. PMID- 2561999 TI - Effector-target co-aggregation as a crucial step in the neutrophil-mediated tumour cell lysis. AB - The Daudi cell lysis by human neutrophils, incubated with phorbol myristate acetate (PMA), was inhibited by amino acids (taurine, methionine), consistent with the involvement of hypoclorous acid (HOCl) in the lytic process. Also, the lysis was inhibited by a monoclonal antibody (mAb J-90) directed against the leukocyte function-associated antigen-1 (LFA-1). The inhibition of the target cell lysis by mAb J-90 is not due to a HOCl-scavenging mechanism, as suggested by the use of control mAb Dako M-1 (anti CD-15). As detected by the microscopic examination of samples from test tubes and measured by monitoring the light transmission from the cell suspensions, neutrophils and Daudi cells were found to co-aggregate during the lytic reaction. Co-aggregation was efficiently inhibited by the mAb J-90. The results suggest that tumour cell lysis by PMA-triggered human neutrophils involves at least two events: production of HOCl and LFA-1 mediated effector-target adhesion. PMID- 2562000 TI - Mechanisms of B-lymphocyte suppression induced by acute physical exercise. AB - Blood mononuclear cells from 20 healthy untrained volunteers were isolated before, during, 2 hr and 24 hr after bicycle exercise at 80% of VO2 max for 1 hr. A reverse plaque forming cell assay was used to investigate B-lymphocyte function. Stimulation with pokeweed mitogen, recombinant interleukin 2 and Epstein-Barr virus resulted in significant decreases in numbers of IgG-, IgM- and IgA-secreting blood cells during as well as 2 hr after exercise, with reversal to pre-exercise values 24 hr later. During and after physical activity we found an unchanged concentration of CD20-positive B lymphocytes suggesting that the suppression of immunoglobulin secreting cells was not due to changes in numbers of B lymphocytes. A decline in CD4/CD8 ratio was measured only during exercise with normalization after exercise. Therefore the B-lymphocyte suppression, most pronounced 2 hr after exercise, was presumably not due to changes in T lymphocytes also indicated in the experiments using EBV-stimulated cultures since EBV acts directly on B lymphocytes. 2 hr after physical activity an increased level of CD14-positive monocytes were observed and the monocytes expressed higher levels of surface HLA-DR during as well as 2 hr after exercise. Addition of indomethacin caused an increased response only in the IL-2 stimulated cultures, suggesting that IL-2-sensitive activated B lymphocytes are downregulated by prostaglandins. Purified B lymphocytes produced plaques only after EBV stimulation, and in these cultures no exercise-induced suppression was found, likewise suggesting an inhibitory effect of the activated monocytes. PMID- 2562001 TI - Opioid peptides. Partially modified retro-inverso dermorphin analogues. XIII. AB - We studied the effect of partial retro-inverso modification of a selected peptide bond of dermorphin (H-Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH2) related peptides. The drastic loss of mu-receptor affinity following introduction of retro-inverso peptide bond between Phe1-D-Ala2 in the new peptide analogues, emphasized the importance of this backbone in the dermorphin peptides. PMID- 2562002 TI - Viruses and cancer of the immune system. PMID- 2562003 TI - Is magnetic resonance imaging more sensitive than computed tomography in the location of corticotropin secreting pituitary adenomas? AB - Two cases of pituitary-dependent Cushing's syndrome are described in which the computed tomography (CT) examination was negative; as the hormone dynamic investigations were directed towards the presence of corticotropin (ACTH) secreting pituitary formations, magnetic resonance imaging (MRI) of the pituitary was performed, which evidenced the presence of such lesions; subsequent neurosurgery confirmed in both cases the location indicated by MRI. In conclusion, the higher sensitivity of MRI compared to CT in the diagnosis of ACTH secreting pituitary adenomas can be noted. PMID- 2562004 TI - [Epidemiological evaluations of human immunodeficiency virus, herpes simplex virus type 1 and 2 and cytomegalovirus infections in drug addicts]. AB - Eighty-eight drug addicts from the "BAN Center" in Torre Annunziata (Naples) and 88 normal subjects pair-matched for age and sex were tested for IgG to human immunodeficiency virus (HIV), herpes simplex virus (HSV) type 1 and 2 and cytomegalovirus (CMV). A high prevalence of subjects with antibodies to HSV-1 and CMV (80.7% and 65.9%) were recorded in the control group testifying to the high level of these infections in Campania. Prevalences were higher in drug addicts, and drug abuse was identified as a risk factor for the acquisition of CMV infection (odds ratio = 2.3). Moreover, drug addiction is also a risk factor for HSV-2 and HIV infection as demonstrated by the observation that drug abusers were anti-HSV-2 (9.1 vs. 1.1%, odds ratio = 6.16) or anti-HIV (11.4 vs. 0%, odds ratio = 23.6) positive more frequently than normal controls. Thus, drug addiction is a risk factor for the acquisition of HIV, HSV-2 and CMV infections. This is probably due to similar habits, frequent among drug addicts from our geographic area and uncommon in the normal population, such as tattooing, needle-sharing needlestick and unsafe sex. Some of these habits, such as unsafe sex and tattooing, seem to be, per se, risk factors for the acquisition of both HIV and CMV infections. The data also suggest that HIV infection was probably introduced in Campania more recently than in northern and central Italy where the prevalence of anti-HIV positive cases among drug addicts is definitely higher. PMID- 2562005 TI - Phosphorylation of HeLa cell multiprotein DNA polymerase alpha complex: impact on activity and partial purification of the associated kinase. AB - Phosphorylation is a major post-translational regulatory mechanism and plays a key role in transduction of mitogenic signals in cell proliferation. The role of phosphorylation and dephosphorylation in regulating the activities of a multiprotein DNA polymerase alpha complex was examined. Treatment of the HeLa cell multiprotein DNA polymerase alpha with calf intestinal alkaline phosphatase resulted in the inactivation of DNA polymerase alpha and DNA primase but had no effect on deoxyribonuclease- and primer-recognition proteins. A protein kinase co purified with the multiprotein DNA polymerase alpha and was partially purified from HeLa cells. The partially purified kinase was active in phosphorylating dephosphorylated polymerase alpha and used casein and histones as exogenous substrates. This study demonstrates that phosphorylation-dephosphorylation may have modulated the activities of DNA replicative enzymes and suggests a role for specific phosphatases and kinases in this process. PMID- 2562006 TI - Cellular pharmacology of DUP-785, a new anticancer agent. AB - DUP-785, a new inhibitor of dihydroorotate dehydrogenase, is currently undergoing clinical evaluation for anticancer activity. We developed a GC/MS method to quantitate dihydroorotate that accumulates in cultures of L1210 cells exposed to growth inhibitory concentrations of DUP-785. This method was used to follow the onset, extent, and duration of inhibition of de novo pyrimidine synthesis in intact L1210 cells and to compare this inhibition with cell proliferation and cellular concentrations of pyrimidine nucleotides. There were direct relations between inhibition of de novo pyrimidine synthesis, changes in pyrimidine nucleotide concentrations, and cell proliferation following short (less than 24 hr) drug exposures; with prolonged exposures (greater than 24 hr), however, there was a departure from these relationships in that restoration of pyrimidine nucleotide pools and de novo pyrimidine pathway activity did not restore cell proliferation. Exposure of L1210 cells to 15 microM DUP-785 produced a maximum cell kill (99.9% as determined by cloning efficiency) at 24 hr, and no increase in cell kill was observed with drug exposure up to 96 hr. PMID- 2562008 TI - Immunologic aspects of myositis. PMID- 2562007 TI - Camptothecin hypersensitivity in poly(adenosine diphosphate-ribose) polymerase deficient cell lines. AB - Mutant cell lines, derived from the Chinese hamster V79 cell line, deficient in poly(adenosine diphosphate-ribose) polymerase activity, and previously shown to be resistant to topoisomerase II inhibitors, were found to be hypersensitive to camptothecin, a topoisomerase I inhibitor. In all the cell lines, camptothecin induced dose-dependent protein-associated DNA single-strand breaks and sister chromatid exchanges. The increased sensitivity to camptothecin-induced cytotoxicity was not associated with an increase in DNA single strand breaks or sister chromatid exchanges. These results suggest the absence of any direct causal relation between (1) camptothecin induced sister chromatid exchanges and cytotoxicity or (2) camptothecin induced DNA strand breaks and cytotoxicity. The hypersensitivity of these mutant cell lines to camptothecin suggests that poly(adenosine diphosphate-ribose) polymerase is involved with topoisomerase I in modulating camptothecin induced cytotoxicity. PMID- 2562009 TI - Inclusion body myositis. PMID- 2562010 TI - [Role of low molecular weight heparin in the prevention of postoperative deep venous thrombosis. Our experience in 88 cases]. AB - In Leriche's "maladie post-operatoire" one of the most frequent and dangerous complications is the deep vein thrombosis (DVT). In general surgery its occurrence is between 10 and 40%, closely related to factors like patient's age, postoperative immobilization, type and time of surgery. Low molecular weight heparins (LMWH) have been recently introduced as a therapy to prevent postoperative DVT in high risk patients. LMWH seem to have all advantages of classical heparin products without their side effects. Eighty eight patients who underwent general surgery were studied in a clinical controlled trial about effectiveness of a new LMWH (Fluxum) for the prevention of postoperative DVT: 44 were treated with Calcium-heparin and 44 were treated with Fluxum. Both drugs decreased significantly the percentage of DVT, but Fluxum seemed to be better tolerated for its single-daily subcutaneous administration. PMID- 2562011 TI - [Primary malignant tumors of the liver]. PMID- 2562012 TI - Structural chromosome aberrations in lymphocytes from children previously treated for Wilms' tumor or Hodgkin's disease. AB - Nineteen children treated for Wilms' tumor (thirteen cases) or Hodgkin's disease (six cases) with cytostatic agents and/or radiotherapy were studied cytogenetically on lymphocytes cultivated from blood samples drawn after at least 1 year of complete remission after end of therapy. A reference group of children was matched for age, sex, and residence. The frequencies of sister chromatid exchange (5.4 versus 5.6 SCE/cell), and chromosome damage type gaps (6.6 versus 7.1%) and breaks (1.9 versus 1.9%) were not different in the two groups, but exchange type aberrations were more frequent in the patients (0.9 versus 0.06%). Fifty karyotypes were analyzed in all but two cases of Hodgkin's disease. The overall frequency of stable (3.1 versus 3.8%) and unstable (1.7 versus 1.4%) structural chromosome changes such as translocations, deletions, chromatid exchanges, and dicentrics were not different in the patient and the control groups. If the chromosome data reflect a general cancer risk, this risk cannot be considerably higher among the cancer-treated children. PMID- 2562013 TI - Urography versus DMSA scan in children with vesicoureteric reflux. AB - Following the diagnosis of primary vesicoureteric reflux, identified as part of the investigation of urinary tract infection, 299 refluxing kidneys in 202 children (aged 0-14 years) were prospectively evaluated using intravenous urography (IVU) and the DMSA renal scan at least 4 weeks after urine infection. There was 88% concordance between IVU and the DMSA scan, but in 12% there were discrepancies manifested in 37 kidneys from 31 children. Thirty-four kidneys were normal on IVU but showed scars of reflux nephropathy (RN) on the technetium 99m- dimercaptosuccinic acid (DMSA) renal scan; 4 of these (2 infants and 2 pre-school children) had severe generalized changes on scanning. Three kidneys were normal of DMSA scan and, although abnormal on initial IVU, were considered to be normal when this was repeated. During a follow-up period of 5 years an annual DMSA was undertaken in 194 patients and the renal scars remained unchanged in all except 1 child. The IVU was repeated 1-3 years after the initial study in 31 children in which the results of the first imaging did not agree. In 28 patients (34 kidneys) in which the initial IVU was normal but the DMSA abnormal, IVU evidence of scarring emerged in 30 of 34 kidneys, including the 4 patients with severe generalized damage on the DMSA. We conclude that abnormalities detected by the DMSA scan may precede the radiological findings, especially in young children. Even severe RN can be established in kidneys that appear normal on the IVU. PMID- 2562014 TI - The DMSA scan and intravenous urography in the detection of renal scarring. PMID- 2562015 TI - Renal tubular differentiation in mouse and mouse metanephric culture. II. Na-K ATPase activity. AB - Sodium-potassium adenosine triphosphatase (Na-K-ATPase) activity was measured by microassay, and the surface density of basolateral membranes was measured morphometrically in postglomerular segments of single tubules isolated from normally developing, intact mouse kidneys and from transfilter metanephric cultures. Proximal tubule Na-K-ATPase activity was 1092 +/- 480 pmol/mm per hour in newborn mice, increasing to 2462 +/- 258 in 1-week-old and 3470 +/- 578 pmol/mm per hour in adult mice. The Na-K-ATPase activity in newborn mice was approximately one-third of the activity in adult mice. Tubular Na-K-ATPase in transfilter metanephric culture was 972 +/- 536 pmol/mm per hour, a mean value almost identical to that in newborn mice. The surface density of basolateral cell membranes was 1.36 +/- 0.60 microns2/microns3 in newborn mice and 1.34 +/- 0.45 microns2/microns3 in 1-week-old mice, increasing to 2.70 +/- 0.98 microns2/microns3 in 4-week-old mice and 2.89 +/- 0.51 microns2/microns3 in adult mice. The surface density of tubular basolateral cell membranes in transfilter metanephric culture was 1.13 +/- 0.51 microns2/microns3, not significantly different from the surface density in newborn mice. The calculated mean surface area of basolateral membranes per unit tubular length was greater in cultures than in newborns, however, because total epithelial volume per unit length was significantly larger in the cultured tubules. Membrane surface area in intact mice increased with age, the surface area per unit length of tubule in adults being 4.6 times the area in newborn animals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562016 TI - Studies of terminal differentiation of electrolyte transport in the renal proximal tubule using short-term primary cultures. AB - There are several lines of indirect evidence suggesting that the renal tubule cells have not yet reached terminal differentiation at birth. Methods used in cell biology can now be applied to study renal ontogeny. This review describes how primary cultures of proximal tubule cells from rats can be used to investigate developmental changes in Na permeability and Na-K-ATPase-mediated transport. PMID- 2562017 TI - Effects of prenatal exposure to ethanol on rat liver development. PMID- 2562018 TI - Effect of cuprophan, hemophan and polysulfone membranes on the oxidative metabolism, degranulation reaction, enzyme release and pulmonary sequestration of granulocytes. PMID- 2562020 TI - [Implantation of porous hydroxyapatite into periodontal bony defects]. AB - The purpose of the present report was to evaluate on a clinical and histological level the effect of hydroxylapatite (HA)--Interpore 200--implanted into various types of periodontal defects. Additionally in-vitro studies with enzymatically released calvarial cells from fetal rats were performed in the organoid culture system. The clinical data 12 months after surgery revealed a greater reduction of probing depths (PD) and gain of attachment level in the implanted sites (59 sites) when compared to the sites treated by open flap curettage (40 sites). When the results were analyzed according to the initial PD the measurements showed no significant difference between the two modalities of treatment in moderately deep pockets (PD less than or equal to 6 mm) in contrast to deep and extremely deep pockets (PD = 7-9 mm and PD greater than 9 mm). In order to collect data about the type of periodontal wound healing biopsies, respectively block sections were taken 6-12 months after therapy in 6 patients and evaluated by light microscopy. Three forms of tissue reaction in relation to the implants were observed. The first consisted of a fibrous connective tissue encapsulation of the implants, the second showed organized bone tissue around the HA, while the third type of tissue reaction consisted of a deposit of mineralized tissue on the surface limited in width, which was interpreted as cementum formation. Analysis of the block sections revealed new connective tissue attachment to a certain extent and bone/cementum-like tissue formation. The possibility of cementum-like tissue formation on HA was confirmed transmission electronmicroscopically by the in vitro experiments.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562019 TI - Protein adsorption on dialyzer membranes influences their biocompatibility properties. PMID- 2562021 TI - [Interaction between orthodontic tooth movement and hydroxyapatite ceramics]. AB - A pilot study examined the clinical and histological effects of orthodontic tooth movement in hydroxyapatite-augmented ridges in animal models. Responses observed at the interface between the clinical root and the ceramo-osseous regeneration were negative: impairment of periodontal function had led to immobility and root resorption. On the basis of this experience one must advice against using this approach in humans at the present time. Further studies will be conducted to elucidate the relevant clinical questions. PMID- 2562022 TI - [Sex and age differences in beta receptor subtypes of bladder sympathetic nerves in the rabbit determined by receptor analysis]. PMID- 2562023 TI - [Tiropramide action on smooth muscle of the lower urinary tract]. PMID- 2562024 TI - [Changes in isolated uterine smooth muscle cells during hormone loading in rats]. PMID- 2562025 TI - [Action of cytochalasin B on the contraction of smooth muscle of guinea pig taenia coli]. PMID- 2562026 TI - [Effect of calyculin A on the contraction of colonic smooth muscle of guinea pigs]. PMID- 2562027 TI - [Effects of endothelin and receptor agonists on tracheal smooth muscle in rats]. PMID- 2562028 TI - [Effect of 5-HT3 receptor antagonists on gastrointestinal motility]. PMID- 2562029 TI - [Effect of various muscle constrictors and cyclic AMP related relaxants on tension and intracellular concentration of Ca of tracheal smooth muscle]. PMID- 2562030 TI - Summary of notifiable diseases in the United States. PMID- 2562032 TI - Ouabain-resistant (Na+,K+)-ATPase enzyme activity in chemically induced ouabain resistant C3H/10T1/2 cells. AB - To further understand the molecular nature of changes leading to chemically induced ouabain resistance in C3H/10T1/2 Cl 8 (10T1/2) cells, we isolated plasma membranes from wild-type and ouabain-resistant (Ouar) 10T1/2 cells and characterized (Na,K)-ATPase activity in the plasma membrane fraction. (Na+,K+) ATPase enzyme activity in membrane fractions extracted from wild-type 10T1/2 cells was inhibited in a concentration-dependent manner by ouabain and was completely inhibited by 2.4 mM ouabain. Lineweaver-Burke and Eisenthal/Cornish Bowden analysis indicated that the inhibition was uncompetitive. Ten to 45% of (Na+,K+)-ATPase enzyme activity extracted from three Ouar 10T1/2 cell lines cultured in 1 mM ouabain was resistant to 2.4 mM ouabain, depending on the cell line. Resistance of (Na+,K+)-ATPase activity in the plasma membrane fraction of Ouar cells to inhibition by ouabain and resistance of cultured Ouar cells to the cytotoxicity of ouabain occurred over similar concentrations of ouabain (0.1 3mM). Two ouabain-resistant cell lines, Ouar MNNG Cl 2 and Ouar MCA Cl 16-7, demonstrated the same total (Na+,K+)-ATPase specific activity as 10T1/2 cells, but the fraction of Ouar enzyme activity increased (from 18 to 40% in MNNG Cl 2 cells and from 10 to 25% in Sp Ouar Cl 16 cells) when the cells were cultured in ouabain. Thermal denaturation profiles and pH dependence profiles of (Na+,K+) ATPase activity in plasma membranes from wild-type and Ouar 10T1/2 cells were identical. A 3.9-kb (Na,K)-ATPase alpha subunit mRNA transcript was found in 10T1/2 cells, and in the Ouar MNNG Cl 2 cell line cultured in the presence or absence of ouabain. There was no amplification of the gene coding for the alpha subunit of (Na+,K+)-ATPase in the chemically induced Ouar MNNG Cl 2 cell line, whether this cell line was cultured in the presence or absence of ouabain. These studies provide further evidence that the Ouar phenotype of chemically induced and spontaneous Ouar 10T1/2 cell lines derives from Ouar (Na+,K+)-ATPase activity, and that this Ouar (Na+,K+)-ATPase activity increases further in some cell lines cultured in the presence of ouabain. PMID- 2562033 TI - Psychiatric disorders associated with corticosteroid therapy. AB - Psychiatric disorders accompanying corticosteroid therapy are reported. They are much more frequently encountered than is generally believed. Moreover, they seem to correlate with the dosage applied. For more convenient clinical approach, we suggest classification of the disorders into four groups. The authors report two of their cases suffering from multiple sclerosis with associated steroid-induced psychosis. Besides the review of etiological mechanisms, the approach to treatment of psychiatric complications induced by steroid therapy is also suggested. PMID- 2562034 TI - [Receptors for biologically active substances in the light of the studies using monoclonal antibodies]. AB - This paper presents a review of studies on the structure and function of receptors for biologically active substances (hormones, drugs etc.). Application of the methods utilizing monoclonal antibodies are described and discussed. PMID- 2562035 TI - [In vitro studies of the effect of growth factors on the connective tissue]. AB - The paper presents recent data concerning the effect of growth factors on the metabolism of collagen, collagenases and glycosaminoglycans in vitro. The hypotheses of the mitogenic action of peptide factors and their role in the control of synthesis and breakdown of polyphosphoinositides are also reviewed. PMID- 2562036 TI - [Immunological aspects of the treatment of various forms of Alzheimer's disease]. PMID- 2562037 TI - [Fatal adenovirus pneumonia]. AB - Adenoviruses are well known and ubiquitous causes of respiratory illness in children. Serious long-term sequelae and even fatalities have been described in young children affected by adenoviral infections of the respiratory tract. This is a report on clinical and histopathological features of 25 infants dying from acute lower respiratory tract infections apparently caused by adenoviruses, from January 1st 1983 through December 31st 1986. The diagnosis was based on the finding of distinctive intranuclear inclusions in the pulmonary lesions that were present in all 25 cases at necropsy. Necrotizing bronchiolitis was found in 24 patients (96%). Nineteen patients (76%) were younger than 1 year of age and 15 (60%) had underlying malnutrition. In 17 instances (68%) the respiratory illness was nosocomial in origin. A specific etiologic diagnosis was not made in any of the 25 patients during the course of the illness, twenty four of them (96%) received various antimicrobial drugs without success. PMID- 2562038 TI - The isoelectrofocusing technique in comparison of some Sudanese type SAT-1 foot and-mouth disease viruses. AB - Isoelectric focusing technique (IEFT) was employed to compare type SAT-1 FMD virus from Sudan. Results of the IEFT tests were compared with available previous serological and epidemiological data on the viruses used. Possible potential uses of the test, in parallel with previously available serological and epidemiological data, are discussed. PMID- 2562039 TI - Infectious bursal disease (IBD) and coccidiosis concurrent infections in Nigerian indigenous chickens. A case report. AB - In an outbreak of concurrent infectious bursal disease (IBD) and caecal coccidiosis in a flock of 45 Nigerian indigenous chickens comprising of 36 unsexed 29 days old chicks and 9 adult hens, 75 per cent mortality was recorded. The adult hens were not affected. PMID- 2562040 TI - Delta-sleep inducing peptide (DSIP) and its fragments as the modulators of neural transmission in the central nervous system. AB - The present study is a continuation of our previous experiments on DSIP activity which have revealed that nonapeptide DSIP inhibits hippocampal electrical activity of the 4-7 c/s frequency band. The aim of the present study was to find which of the known DSIP fragments is responsible for its activity, i.e. to find the active site of the molecule. The experiments were carried out with the entire DSIP molecule and its three different fragments. The method of threshold continuous arousal pattern (TCAP) monitoring was used as the indicator of DSIP activity. It was found that the entire DSIP molecule increased TCAP, while its 1 5 fragment decreased it 1-4 and 5-9 fragments had no noticeable effect. PMID- 2562041 TI - Genetic alterations and growth factors in the pathogenesis of von Recklinghausen neurofibromatosis. AB - This paper is a summary review of recent developments with regard to the pathogenesis of von Recklinghausen neurofibromatosis, or NF-1. In the section on molecular biology of NF-1, the focus is on genetic linkage, with a note about prenatal diagnosis, tumor genetics and animal models. In the section on growth factors and receptors, the first element deals with neurofibromatosis as a model of cellular interaction, and particularly the issues of dysplasia versus neoplasia, cell-cell interactions and 'vicious cycle' positive feedback mechanisms; additional elements focus on nerve growth factor-beta, Schwann cell mitogens and other growth factors. In the last section, the focus is on further developments in terms of molecular biology, but with additional emphasis on the need for very intense input from the clinicians characterizing the many phenotypes of NF-1 and its NF and non-NF alternatives. PMID- 2562042 TI - Manganese accumulation in yeast cells. Electron-spin-resonance characterization and superoxide dismutase activity. AB - Manganese accumulation was studied by room-temperature electron spin resonance (ESR) spectroscopy in Saccharomyces cerevisiae grown in the presence of increasing amounts of MnSO4. Mn2+ retention was nearly linear in intact cells for fractions related to both low-molecular-mass and macromolecular complexes ('free' and 'bound' Mn2+, respectively). A deviation from linearity was observed in cell extracts between the control value and 0.1 mM Mn2+, indicating more efficient accumulation at low Mn2+ concentrations. The difference in slopes between the two straight lines describing Mn2+ retention at concentrations lower and higher than 0.1 mM, respectively, was quite large for the free Mn2+ fraction. Furthermore it was unaffected by subsequent dialyses of the extracts, showing stable retention in the form of low-molecular-mass complexes. In contrast, the slope of the line describing retention of 'bound' Mn2+ at concentrations higher than 0.1 mM became less steep after subsequent dialyses of the cell extracts. This result indicates that the macromolecule-bound Mn2+ was essentially associated with particulate structures. In contrast to Cu2+, Mn2+ had no effect on the major enzyme activities involved in oxygen metabolism except for a slight increase of cyanide resistant Mn-superoxide dismutase activity, due to dialyzable Mn2+ complexes. PMID- 2562043 TI - [A case of malignant retroperitoneal fibrohistiocytoma]. AB - A case of retroperitoneal malignant fibrous histiocytoma, with particular reference to its clinico-pathologic characteristics, is reported. The primary large lesion, presenting with a cavitary lung metastasis, resulted to be unresectable at surgery because of major abdominal vessels infiltration. PMID- 2562044 TI - [Mucoid carcinoma of the anus and perianal fistula: a clinical case and review of the literature]. AB - The Authors describe a case of mucoid anal carcinoma occurring in a perianal fistula. Surgical treatment was an abdomino-perineal resection (Miles' operation). On the basis of their experience and of the results reported in the literature, the Authors discuss the clinical and histological aspects of this association. PMID- 2562045 TI - [Ultrastructural and immunoelectron microscopic study of the desmoplastic stroma in carcinoma of the breast]. AB - Different collagen types (I, III, IV, V) were identified in breast carcinoma desmoplastic tissue by ultrastructural analysis and immunoelectronmicroscopy. Type V collagen is present as a 12 nm. fibril in the stroma, either adjacent to the basement membrane or concentrated around the thicker fibers. Myofibroblasts, fibroblasts and tumor cells can be its major producers. Its possible function as a bridge between different collagens can be utilized, with different finality, by the same cells that have produced it. Furthermore, type V collagen can be involved in tumor invasion of the stroma and in all directional movements of tumor cells, as already demonstrated for other extracellular matrix components. PMID- 2562046 TI - Suppression of immunity to influenza virus infection in the respiratory tract following sleep disturbance. AB - The extent to which sleep deprivation interferes with immunity in the respiratory tract to influenza virus has been assessed in mice. Mice were orally immunized with influenza virus on two occasions separated by a one week interval and challenged intranasally one week later. Some animals were deprived of sleep for a 7 h period immediately following challenge. Three days after challenge, virus clearance and virus specific antibody were determined in lungs of sleep deprived and normally sleeping mice and the results compared with unimmunized mice subjected to the same protocol. Whereas immunized, normal sleep mice achieved total virus clearance, sleep deprivation in immunized mice completely abrogated this effect such that sleep deprived animals behaved as though they had never been immunized. There was no difference in viral clearance in unimmunized mice whether sleep deprived or not, indicating that sleep deprivation did not itself have a direct effect on viral replication. The data reported here support the concept that sleep is a behavioral state which is essential for optimal immune function in the presence of a respiratory tract pathogen. PMID- 2562047 TI - Studies on the effect of monoamine antagonists on the morphogenesis of the newt. AB - The effects of three monoamine antagonists, p-chlorophenylalanine, diethyldithiocarbamate and propranolol on the morphogenesis of newt embryos were studied. Antagonists were administered during late blastula through neurula stages. In a concentration of 1 mM, all three arrested gastrulation and caused disintegration of the embryos. Lower concentrations (0.1-0.5 mM) retarded morphogenetic movements in the gastrulation and caused malformations especially in the anterior parts of the embryos; pigmentation was delayed by 1 or 2 days. In addition, p-CIPhe inhibited yolk granule degradation in the notochord and DEDTC caused notochordal hypertrophy. The results show that interference with synthesis or action of catecholamines and serotonin affects morphogenesis. With the methods used it is not possible to discover exactly how monoamines regulate the morphogenetic events because of the unspecific side effects of the antagonists and the feedback interactions between the monoamines. PMID- 2562048 TI - Neural induction. PMID- 2562049 TI - [Calcitriol resistant rickets]. PMID- 2562050 TI - [Metastases of lung carcinoma to the tongue]. AB - Metastatic tumors in soft tissues of oral cavity are uncommon, but tongue metastasis are quite infrequent. If it occurs, the invasion is indicative of a drop in the whole organism defenses. We describe two cases of lung carcinoma spreaded to the tongue and the post-mortem examinations. Before oral manifestations we confirm the development of many metastases located in various organs. PMID- 2562051 TI - Dopaminergic involvement in nociceptive sensitivity/behavioral reactivity regulation during aversive states of different nature in the rat. AB - To investigate the involvement of dopamine (DA) in nociceptive sensitivity behavioral reactivity regulation in animals during aversive states of different nature, the influence of pharmacologically-induced decrease and increase of DA neurotransmission on vocalization and movement reactivity were studied in rats in free behavior, during restraint stress, after acute trauma of an extremity and under intraperitoneal acetic acid administration. The influence of longterm increase (apomorphine in a high dose) and decrease (haloperidol, apomorphine in a low dose) on suprarenals weight and gastric ulceration in animals exposed by polymodal aversive stimulation was also studied. The data obtained are discussed in relation with; 1. DA involvement in regulation of nociceptive sensitivity and behavioral reactivity in aversive environment; 2. the role of DA and endogenous opioid peptides in endogenous analgesic mechanisms; 3. the functional significance of cerebral DA in organization and realization of various types of an organism's adaptive activity produced by different environmental and homeostatic variables; and 4. the interaction of DA and endogenous opioid peptides in mediation of this activity. PMID- 2562052 TI - Alpha melanocyte stimulating hormone (MSH) in Tourette's syndrome. AB - The intermediate lobe of the pituitary is thought to be the primary source of alpha-melanocyte stimulating hormone (alpha-MSH). While the intermediate lobe of the human fetus contains high concentrations of alpha-MSH, the human adult pituitary is devoid of the peptide. Based on evidence implicating the hypothalamus as a site of dysfunction in Tourette's syndrome (TS), and studies suggesting that alpha-MSH may be involved in neuronal maturation, we measured plasma alpha-MSH levels in three unmedicated TS patients (aged 12 to 69), and in three age-matched controls. High plasma alpha-MSH concentrations were detected in all three patients. None of the patients showed any abnormalities in skin coloration. These findings suggest that abnormalities in the synthesis or release of alpha-MSH may be linked to the pathophysiology of TS, and also to the delay in neurodevelopmental maturation. PMID- 2562053 TI - Cortical and retinal refractory periods in the human visual system. AB - Refractory periods of the visual system were investigated in 12 healthy subjects by simultaneously recording retinal (ERG) and cortical (VEP) evoked electrical activity. Double-flash stimuli were presented at different interstimulus intervals, and response components evoked by the second flash were analyzed in detail, and related to psychophysical detection thresholds. With short interstimulus intervals ERG b-wave peak latencies were increased and b-wave amplitudes were significantly reduced, while P100 component latencies of the VEP were significantly influenced only at long interstimulus intervals. Regression analysis of the individual data as well as analysis of the retinocortical transmission times showed that the cortical latency changes were not simply caused by changes on the level of the retina. Additional influences of the interstimulus interval on nonretinal structures of the human visual system must be assumed. The subjective psychophysical detection thresholds were significantly higher than the threshold values at which reliable electrical or cortical response components could be elicited. PMID- 2562054 TI - Hormonal control of Leydig cell function. Molecular endocrinology section, endocrinology and reproduction. PMID- 2562055 TI - [Is renal function of hydronephrotic kidneys reversible]. PMID- 2562056 TI - Ca2(+)-mediated degradation of central nervous system (CNS) proteins. PMID- 2562057 TI - Voltage-dependent GABA-induced modulation of calcium currents in chick sensory neurons. AB - Externally applied gamma-aminobutyric acid (GABA) quickly and reversibly reduces by 60% voltage activated Ca2+ currents in chick dorsal root ganglion cells. This action is antagonized by depolarization, with characteristic time and voltage requirements. Intracellular perfusion with guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) or guanosine 5'-O-(2-thiodiphosphate) (GDP beta S) mimicks and blocks the GABA effect, respectively. A 3-state model describing the reactions involved is proposed. PMID- 2562058 TI - Presynaptic calcium currents recorded from calyciform nerve terminals in the lizard ciliary ganglion. AB - Electron microscopic examination of ciliary ganglion from Anolis carolinensis shows large calyciform presynaptic nerve terminals ending on ganglion cells. Intracellular records were obtained from the terminals, and membrane currents were recorded with the single-electrode voltage clamp technique. After block of Na+ currents with tetrodotoxin, depolarization of the terminals produced inward currents that disappeared when extracellular Ca2+ was removed, and increased in magnitude and duration when competing outward currents were blocked by intracellular Cs+. Thus it is possible, with this preparation, to record and characterize Ca2+ currents presumably associated with neurotransmitter release. PMID- 2562059 TI - Effects of hypoxia on cyclic nucleotide formation in rabbit carotid body in vitro. AB - The present experiments measured cAMP and cGMP in the arterial chemosensory tissue of the rabbit carotid body exposed for 10 min in vitro to normoxic or hypoxic conditions, or to specific activators of adenylate cyclase (forskolin) and guanylate cyclase (sodium nitroprusside). The enzyme activators elevated the basal levels of cAMP (48 x) and cGMP (3.7 x), respectively. Hypoxic media increased cAMP in the carotid body by 3.6-fold, but the levels of cGMP were reduced by 33% in media equilibrated with low O2. The data are consistent with the notion that cyclic nucleotides are involved in the transduction of natural stimuli and/or the neurotransmitter feedback modulation of chemosensory type I cells. PMID- 2562060 TI - MK-212 increases rat plasma ACTH concentration by activation of the 5-HT1C receptor subtype. AB - The effects of the serotonin agonist MK-212, on rat plasma ACTH were examined. MK 212 significantly increased plasma ACTH levels, and this effect was blocked by the 5-HT1C antagonists mesulergine and metergoline but not by spiperone, ketanserin, or (-)-pindolol. The results suggest that MK-212 activates the 5-HT1C receptor subtype to increase ACTH. PMID- 2562061 TI - Involvement of postsynaptic alpha 2-adrenoceptors and guanine nucleotide-binding protein in guanabenz-induced cardiovascular suppressant effects in the rat. AB - In adult, male Sprague-Dawley rats anesthetized with pentobarbital sodium, pretreatment with the catecholamine-depleting agent, reserpine (150 micrograms, i.c.v.) significantly antagonized the hypotensive and negative inotropic and chronotropic effects of guanabenz, either given intravenously (100 micrograms/kg) or microinjected bilaterally (5 micrograms) into the nucleus reticularis gigantocellularis (NRGC), a medullary site of action for this centrally acting antihypertensive agent. Pretreating animals with microinjection of the selective norepinephrine neurotoxin, DSP4 (50 micrograms), into the bilateral NRGC, on the other hand, did not appreciably blunt the cardiovascular suppressive actions of the aminoguanidine compound. I.c.v. administration of pertussis toxin (2.5 micrograms), which potentially blocks the action of two guanine nucleotide binding proteins (Gi and Go), significantly antagonized the circulatory inhibitory effects of guanabenz (100 micrograms/kg, i.v.). More specifically, this blocking effect was still apparent upon microinjecting pertussis toxin (250 ng) into the bilateral NRGC. These data suggest that both pre- and postsynaptic alpha 2-adrenoceptors, and a pertussis toxin sensitive G-protein(s) (Gi and/or Go), in the NRGC are crucial to the expression of the cardiovascular suppressant actions of guanabenz. PMID- 2562062 TI - Antagonist drug selectivity for radioligand binding sites on voltage-gated and N methyl-D-aspartate receptor-gated Ca2+ channels. AB - Drugs that block voltage-gated Ca2+ channels or N-methyl-D-aspartate receptor gated channels have been shown to reduce experimental hypoxic-ischemic neuronal injury. To determine if any such compounds interact with both types of channels, and might therefore be prototypes for new anti-ischemic drugs with dual therapeutic actions, we compared the affinities of channel blockers for voltage gated Ca2+ channel binding sites labeled by (+)-[3H]PN 200-110 and N-methyl-D aspartate receptor-gated channel sites labeled by [3H]MK-801. Combined effects were most prominent with dextromethorphan, followed by D-888, verapamil and dextrorphan. PMID- 2562063 TI - The 24-h rhythm of metabolic activity of the cockroach circadian pacemaker. AB - In the cockroach, Leucophaea maderae, the optic lobes contain a circadian pacemaker. The metabolic activity of optic lobe neuropils was measured at 17 time points over a 24-h period in constant darkness, using cytochrome oxidase activity as functional marker. The results revealed a clear 24-h energy metabolism rhythm in neuropils and particularly in the lobula plate, a part of the region which is assumed to include a circadian pacemaker in this species. PMID- 2562064 TI - Changes in sarcolemmal adenosine triphosphatase activity and in ouabain sensitivity of rat myocardium in isoproterenol-induced cardiac hypertrophy. AB - Sodium, potassium-activated adenosine triphosphatase (ATPase) activity and the sensitivity of rat myocardium to ouabain was studied in isoproterenol (IPR) induced cardiac hypertrophy. IPR in a dose of 5 mg/kg was administered to rats intraperitoneally, once daily, for seven days. Left ventricular trabeculae originating from IPR-treated rats were significantly less sensitive than controls to ouabain-induced positive inotropy. In crude homogenate and sarcolemmal fractions the ATP hydrolysing activity both in the presence of Mg++ (basic) and Mg++, Na+, K+ (total) was significantly reduced in the heart of IPR-treated rats. The difference between the total and basic ATPase, i.e. the Na+, K(+)-stimulated portion of the activity was slightly reduced, but the sensitivity of Na+, K(+) ATPase to ouabain remained unchanged. The results indicate that the well-known relation of sodium pump inhibition to positive inotropy in the heart of IPR treated rats may not be valid. PMID- 2562065 TI - Down-regulation of the GABA-ergic system in selected brain areas of spontaneously hypertensive rats (SHR). AB - The relevance of GABA-ergic system in hypertensive state has been studied. GABA content, GAD activity and GABA-A receptor binding in various brain areas in age matched spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto rats (WKY) was compared. Out of 9 brain areas studied the GABA content was significantly lower in the substantia nigra, hypothalamus, hypothalamus posterior and hippocampus of SHR rats. GAD activity was lowered in the hypothalamus and hippocampus of young SHR and adult SHR rats (4, 8, 14 weeks old). Scatchard analysis of the binding isotherms indicated a lower Bmax of the binding sites in the hypothalamus and hippocampus of 8 and 14 weeks old SHR rats. These results suggest that activity of GABA-ergic system differs substantially in SHR and WKY rats brain. Furthermore, these differences appear already in young prehypertensive SHR rats as well as in the early stages of hypertension. PMID- 2562066 TI - The responsiveness of central benzodiazepine and GABA(A) receptors in vasopressin and spontaneously hypertensive rats. AB - Administration of diazepam, an agonist of benzodiazepine receptors, and muscimol, an agonist of GABA(A) receptors, resulted in an increase in cytosol GABA-modulin in hypothalamus and cerebellum. Low dose of muscimol, ineffective by itself, completely antagonized the isoniazid, bicuculline and picrotoxin-induced decrease in GABA-modulin activity. In contrast, low dose of diazepam was able to block only the action of isoniazid. That confirms different mechanism of action of muscimol and diazepam. The increase in cytosol GABA-modulin activity in the anterior and posterior hypothalamus of spontaneously hypertensive rats (SHR) and of vasopressin hypertensive rats required much higher doses of muscimol than in normotensive animals showing subsensitivity of GABA(A) receptors in these brain structures. In contrast, the sensitivities of GABA(A) receptors in the cerebellum and of benzodiazepine receptors in the hypothalamus and cerebellum were equal to hypertensive and normotensive rats. PMID- 2562067 TI - Primary primitive neuroectodermal tumor of the spinal cord associated with neural tube defect. AB - Certain spinal-cord (SC) neoplasms, principally lipomas and dermoid tumors, have been diagnosed in association with characteristic neuroskeletal malformations thought to result from neural-tube defects (NTD). To our knowledge, no such association has been recognized for primary primitive neuroectodermal tumor of the SC (PNET-SC), an SC malignancy which has been reported in only 3 children and 5 adults. We describe here the occurrence and treatment results of PNET-SC in a boy who exhibited several neuroskeletal malformations suggestive of an underlying NTD. By undertaking a comparative analysis of the literature with respect to both the histology and the clinical presentation of this child's tumor, we document an occurrence of an uncommon malignancy, PNET-SC, and identify PNET-SC as another SC neoplasma which may be associated with NTD. PMID- 2562068 TI - [The neurochemical bases of absence-type epilepsy]. AB - The author attempts to introduce several of the present data concerning the neurochemical basis of one of the most enigmatic neurological disturbances, the so-called "petit mal" epilepsy. The present material is based on some of the most recent studies of neurochemistry, and considerations are also made concerning the experimental modelling of "petit mal" epilepsy. PMID- 2562069 TI - Phaeohyphomycosis due to Phialophora richardsiae. AB - Phaeohyphomycosis, an infection characterised by dematiaceous yeast-like cells, hyphae and pseudohyphae in tissue, is an uncommon condition, often affecting immunosuppressed patients. A sixty four year old boat-builder, receiving treatment with prednisone and azathioprine developed multiple cutaneous nodules on the extremities. Histology showed a mixed dermal inflammatory infiltrate with scattered spores and hyphae. Culture revealed two organisms, Phialophora richardsiae and Exophiala jeanselmei. Fluorocytosine was initially given but the organism was found to be resistant. Since side effects have been associated with long term ketoconazole therapy, a less toxic and more potent triazole compound, itraconazole, was used. After three months, the lesions had completely resolved without adverse clinical or biochemical changes. PMID- 2562070 TI - [Effect of kanamycin on the cochleas in guinea pigs]. AB - Kanamycin (400 mg/kg) was injected daily for 5 days in guinea pigs, the changes in CM and AP responses were recorded. The CM threshold was elevated in 34.5% of the animals and the CM amplitude was reduced when compared with normal animals (P less than 0.005). The AP threshold was elevated in 13.8%. The latency of N1 was increased (P less than 0.001) and the amplitude of N1 reduced at 80 dB (S-L) (P less than 0.001).cAMP and cGMP levels in the organ of Corti and stria vascularis spiral ligament decreased, and the decrease correlated with the changes in auditory function. We consider the reduction of cyclic nucleotides in the cochlea may be important in kanamycin ototoxicity. PMID- 2562071 TI - Giant parapox infection in a two year old child. AB - We report the case of a two year old boy who developed a large tumour on his face. The lesion resembled a pyogenic granuloma clinically and histologically. Viral studies indicated a parapox infection and a bovine source was supported on epidemiological grounds. The lesion was removed by shave excision and the area healed without significant scarring. PMID- 2562072 TI - [Effect of naphthoquinone-imines on the production of hydrogen peroxide by Crithidia fasciculata and Leptomonas seymouri]. AB - It has been synthesized a series of 4-aminoisoxazol-1,2-naftoquinones, which inhibit growth and DNA synthesis in Trypanosoma cruzi. This effect was related to quinone induction of oxyradical generation. N-(5-methyl-3-isoxazolyl)-4-amino-1,2 naftoquinone and 2-hydroxy-N-(3,5-dimethyl-1-4-isoxazolyl)-1,4-naftoquinone-4 imine inhibited also growth of Crithidia fasciculata and Leptomonas symouri, two organisms that may prove useful for the assay of trypanocidal drugs. In order to establish the role of oxyradicals for quinone-imine toxicity, the latter redox cycling was demonstrated by a) reversible change in their spectra; b) reduction under anaerobic conditions, and c) quinol oxidation by oxygen. Production of H2O2, measured by the microperoxidase method, was maximal with N-(5-methyl-3 isoxazolyl)-4-amine-1,2-naftoquinone (IVD) (0.29-0.26 nmol/min/mg of cell protein), either with C. fasciculata or L. seymouri. Lesser, though significant activities were obtained with 2-hydroxy-N-(5-methyl-3-isoxazolyl)-1,4 naftoquinone-4-imine (IIID) and 2-hydroxy-N-(3-methyl-5-isoxazolyl)-1,4 naftoquinone-4-imine (IIIE) whereas IIIA and IIIC were inactive. O2-. generation, measured by the adrenochrome method, was induced by naftoquinone-imines with a hydroxyl group at C-2 (IIIE, 2-hydroxy-N-(3,5-dimethyl-4-isoxazolyl)-1,4 naftoquinone-4-i mine (IIIC) and 2-hydroxy-N-(3,4-dimethyl-5-isoxazolyl)-1,4 naftoquinone-4-i mine (IIIA), but no by IVD, with a carbonyl group at C-2. Measurement of catalase and superoxide dismutase in both organisms yielded significant activities, but ascorbate peroxidase, guaiacol peroxidase and bencidine peroxidase were inactive.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562073 TI - [Protection against encephalitis in rats caused by a pathogenic strain of the Junin virus, using peripheral inoculation of an attenuated strain]. AB - Argentine Hemorrhagic Fever manifests itself in man either subclinically or in hemorrhagic or neurological forms, mortality reaching 20%. Although Candid 1 strain is undergoing pilot trials, current therapy still resorts to convalescent serum administration. A neurological model was used to evaluate protection conferred by the attenuated XJC13 Junin virus strain. Newborn rats inoculated intraperitoneally (ip) prove resistant, whereas 8-12 day-old animals infected by intracerebral route with the XJ prototype strain suffer 100% mortality with neurological signs. The aim of this study was to achieve protection in this model and attempt to elucidate the mechanisms involved in resistance. It was observed that the longer the inoculation challenge interval, the greater was the survival percentage. In protected animals, brain viral titres were 3 log lower than in challenged controls, while XJC13 infected unchallenged controls presented low CNS values throughout. Neutralizing antibody levels were not significantly different in experimental versus challenged control groups, ruling out any secondary booster effect on protected rats. Neither the transfer of immunoserum nor of endogenous or exogenous interferon altered mortality. However, when splenocytes from rats infected 10 days previously were transferred prior to XJ challenge, survival was increased to 50%, but there was no gain in protection when cells were treated with antithymocyte serum plus complement. Consequently, protection in this neurological model can be attributed to a cellular immune response. PMID- 2562074 TI - [Inhibition of the pseudorabies virus (Suis herpesvirus 1) by an antiviral agent isolated from the leaves of Melia azedarach]. AB - A partially purified extract from fresh green leaves of Melia azedarach L prevents the replication of Pseudorabies virus (Suid herpesvirus, (PrV)) in Vero cells. Monolayers of VEro cells were fully protected against PrV CPE when an appropriate concentration of the antiviral compound was added to the culture medium after virus adsorption and remained during 72 h. By contrast, if cells were pretreated with the same amount of antiviral during 18 h before infection, CPE produced by PrV was only slightly reduced. The antiviral activity of the compound specifically affect PrV multiplication since it was found that no toxic effects on cells were observed at the concentrations assayed which inhibited virus yield by 99%. The effect or the antiviral on adsorption or penetration steps was investigated. Neither the number or viral particles that penetrate nor the amount of unadsorbed virus were the same in treated and non-treated cultures. In order to determine whether an early of late step of intracellular virus replication was inhibited, the extract was added at different times post infection and the virus yield was determined. The production of infectious virus decreased almost 3 log when the compound was added up to 4 h. p.i. According to these results we postulate that an early stage of virus replication, probably beta-messenger expression is inhibited. PMID- 2562075 TI - [Constitutive expression of human fibroblast interferon gene controlled by SV40 early promoter in CHO cells]. AB - HindII fragment encoding human fibroblast interferon (IFN beta) gene coding sequence was fused at 60 bp downstream from the RNA start site of SV40 early gene to be a constitutive expression plasmid pSVE beta. This recombinant plasmid was transfected into the dihydrofolate reductase (dhfr)-deficient Chinese hamster ovary (CHO) cells together with a selectable dhfr gene. About half of transformants continuously secreted IFN beta into the supernatant without inducement. One of the subclone transformants constitutively produced up to 852U IFN beta/2 X 10(6) cells/ml. 48hr in common medium. PMID- 2562076 TI - [Wilms' tumor in patients with different abnormalities]. AB - We present three cases of patients with Wilms' tumour. The first case showed a bilateral tumour with hemihypertrophy and Beckwith-Wiedemann syndrome. The second one had hemihypertrophy and double left pyelocalyceal system with double renal tumour. The last case had a horse-shoe kidney with a tumour on the right and a left hydronephrosis. The second and third cases had a good evolution. PMID- 2562077 TI - [Prostanoid precursors in the liver, lung and kidney of swine during ontogenesis]. AB - Phospholipids and the levels of dihomo-gamma-linolenic, arachidonic and eicosapentaenoic acids in particular fractions of phospholipids in pig foetal liver, lung and kidney during ontogenesis, were investigated. A permanent rise in dihomo-gamma-linolenic and arachidonic acid content in phospholipid fractions of the above mentioned pig foetal internal organs was observed. In eicosapentaenoic acid level no changes were noted. These observations are parallel with increased prostanoid production during ontogenesis. The constant level of eicosapentaenoic acid supports the finding, that this acid does not convert to prostanoids in pig smooth muscle cells. PMID- 2562078 TI - [Biochemical properties of Polish and standard strains of Trichophyton equinum]. AB - It was found that strains of T. equinum which were tested, produced in vitro urease, gelatinase, protease for bovine serum, hemolysins for horse and sheep red blood cells, and keratinase for horse, bovine and canine hair. The ability to produce elastase and caseinase was not recorded. In urease test, it was noticed that after three passages on media with urea the time of appearance of positive reactions was shortened and all strains of T. equinum reacted positively during three days. In these studies some differences in enzyme production between T. equinum and T. mentagrophytes were recorded. Generally, T. mentagrophytes revealed stronger enzymatic properties because it produced the enzymes in two ranges of temperature, i.e. 26-28 degrees C and 37 degrees C, and in addition it dissolved horse sera. The lipase production by T. equinum in optimal temperature of growth 26-28 degrees C only, may be used as an additional test for differentiation between T. equinum and T. mentagrophytes. In penetration test, it was found that strains of T. equinum which were tested perforate in vitro horse, bovine and canine hairs but they do not attack human hair. Some differences were seen concerning the way of penetration of horse hair by the fungi. T. equinum produces rectangular and coaxial perforation organs but T. mentagrophytes produces coaxial ones only. PMID- 2562079 TI - [Effect of dopamine on small-intestinal motility in sheep]. AB - The receptors involved in dopamine effect on the motility of the ileum were studied. The experiments were carried out on 3 sheep with chronic fistulas of the ileum and rumen. The contractions of the fistulated parts of the alimentary tract were recorded by the balloon method. Dopamine infused intravenously at a rate 28 38 micrograms.kg-1.min-1 inhibited the motility of the ileum. Domperidone (0.4 0.8 mg.kg-1) did not abolish this inhibiting influence of dopamine. Dopamine infused after propranolol (0.5-0.7 mg.kg-1) inhibited the motility of the ileum, however administrated after phentolamine (0.6-0.8 mg.kg-1) did not change its movements. It has been concluded that dopamine inhibits the ileum motility in sheep through alpha-adrenergic receptors. PMID- 2562080 TI - [Effect of cysteinyl leukotrienes on uterine strips from pregnant and non pregnant swine]. AB - Cysteinyl leukotrienes C4, D4 and E4 elicited concentration-dependent contractile response of pregnant and non-pregnant pig uterine strips during incubation in De Jalon solution in concentrations between 1 x 10(-9)M and 1 x 10(-5)M for LTC4 and LTD4, and between 1 x 10(-8)M and 1 x 10(-5)M for LTE4. The maximum contractions elicited by leukotrienes were 77.5 +/- 3.0%, 86.0 +/- 2.6% and 37.5 +/- 2.5% of these elicited by histamine 1 x 10(-5)M for LTC4, LTD4 and LTE4, respectively. FPL 55712, leukotriene end-organ antagonist, antagonized the uterine contractile response to cysteinyl leukotrienes in dose-dependent manner. Indomethacin, an inhibitor of cyclooxygenase pathway of arachidonic acid metabolism also antagonized leukotriene-induced contractions of pig uterus. Pregnant uteri were more susceptible to leukotriene action than non-pregnant uteri, and response increased parallelly with advancement of pregnancy. These results demonstrate that LTC4, LTD4 and LTE4 possess significant uterine contractile activity in domestic pig, which may partially be mediated indirectly via cyclooxygenase products of arachidonic acid metabolism. PMID- 2562081 TI - Regulation of the erythropoietin gene. PMID- 2562082 TI - [The use of hydroxylapatite in maxillofacial surgery]. AB - The calcium phosphate system, and in particular Hydroxylapatite, has been the subject of intensive investigation. Recently dense, non-resorbable particulate and block forms of hydroxylapatite have been developed for use as tooth root, bone augmentation and bone replacement material. Dense and porous hydroxylapatite forms have been used clinically for periodontal defects, alveolar ridge maintenance and alveolar ridge deficiencies. In this study the different clinical uses of hydroxylapatite in the Maxillofacial area are presented. PMID- 2562083 TI - Decreased GABA B receptors in helpless rats: reversal by tricyclic antidepressants. AB - Recently, sufficient evidence has accumulated to suggest that a central GABAergic dysfunction may be primarily related to the physiopathology of affective disorders and that antidepressant mechanisms have an intrinsic GABAergic component. In depressed patients GABA levels are reported to be low in the cerebral spinal fluid and plasma, and GABA synthesis is decreased in some brain areas, including the frontal cortex. GABA mimetics exhibit antidepressant-like actions in behavioral models in the olfactory bulbectomized rat and in the learned helplessness paradigm. In the olfactory bulbectomized rat, GABA B receptors are down regulated in the frontal cortex and in the learned helplessness paradigm, GABA release is diminished in the hippocampus. These decreases are reversed by antidepressants in parallel with their behavioral activities. In this study, data obtained indicate that in the learned helplessness paradigm, GABA B receptors are decreased in the frontal cortex and this decrease is reversed by imipramine and desipramine (16 mg/kg/day) in animals which are considered to be 'responders' to antidepressant treatments. PMID- 2562084 TI - Cardiac extension of Wilms' tumor. AB - Two children who were brought to the Istanbul Children's Hospital with congestive heart failure caused by extension of Wilms' tumor to the right atrium are presented. In both cases a large mass was noted in the right atrium by two dimensional echocardiography. The tumors were successfully removed at open heart surgery, and chemotherapy and radiotherapy were started postoperatively. The patients are both alive and symptom-free; one, three and a half years and the other two years postoperatively. PMID- 2562085 TI - Prophylactic cranial irradiation for small-cell lung cancer: long-term results. AB - Over a six-year period, 72 patients with small-cell lung cancer received prophylactic cranial irradiation, as well as combination chemotherapy. There were 58 patients with limited-stage disease. With a minimum of four years follow-up, there are only four survivors, one of whom is affected by a crippling treatment induced neurological deficit, and another has moderate impairment of cognitive functions. Despite cranial prophylaxis, 14 patients died from cerebral metastases, and four others of undiagnosed neurological problems. One other patient, who died apparently in remission, also had severe iatrogenic brain damage. These results cast doubt upon the value of this form of treatment in a disease that remains incurable for the vast majority of sufferers. PMID- 2562086 TI - Flow cytometry developments, uses of monoclonal antibodies and cell purification. PMID- 2562087 TI - The effect of pH adjustment of 2% mepivacaine on epidural anesthesia. AB - Two-hundred men, scheduled for elective meniscectomy under epidural anesthesia, were randomly assigned to receive either a standard 2% mepivacaine solution (n = 100) or a pH adjusted 2% mepivacaine solution (pHAS, n = 100). The pHAS was freshly prepared before the block by adding 0.1 mEq of NaHCO3 per ml of mepivacaine solution. After a test-dose, the anesthetic solution was injected to produce a level of sensory anesthesia to T10. The mean duration of anesthesia and the mean dose of mepivacaine used were comparable between the groups. Patients in the pHAS group showed a significant shortening of onset time in T10 and in S2 segment (p less than .001). Grade 3 motor blockade was achieved in the same number of patients, but a faster motor block was observed in the pHAS group (p less than .05). Regression of both sensory and motor blockade in the two groups was not significantly different. PMID- 2562088 TI - [An attempt of in vitro portal invasion model of hepatocellular carcinoma utilizing permeable collagen membrane]. AB - Hepatocellular carcinoma (HCC) possessed the ability of vascular invasiveness toward hepatic portal vein on the process of progression. This biological character of HCC can influence the patients survival on clinically. In this paper, we tried to establish the in vitro portal invasion model with human materials. The hepatic portal vein endothelial cell (HPVEC) derived from intrahepatic portal veins by surgically, have been propagated, as outgrowth cultures in RPMI-1640 medium with 10% fetal bovine serum, on permeable collagen membranes (KOKEN, Tokyo) containing mainly type I collagen, covered with a solubilized tissue basement membrane (MATRIGEL, Collaborate Res., Inc., Bedford MA) involving type IV collagen, laminin and proteoglycan. The primary cultured HPVEC with polygonal shaped cells forming a pavement stone sheet, were positively stained with Factor VIII related antigen and synthesized both prostacyclin and collagenase inhibitor. Co-culture of primary human HPVEC and HuH-7 (human HCC cell line obtained from Prof. Satoh, Okayama Univ.,) cells were inoculated onto reverse side between collagen membrane and gell formed basement membrane. Morphological alterations on the side of HPVEC can be obtained such as polylayered cells and different cytoplasmic cells among HPVEC. These results indicate that this experimental model can provide an useful in vitro model for the study of HCC portal invasion. PMID- 2562089 TI - [Establishment and characterization of a human malignant fibrous histiocytoma line serially transplanted in nude mice]. AB - Serial heterotransplantation of human malignant fibrous histiocytoma (MFH) derived from tibia was attempted in BALB/c nu/nu mice, and HKMFH-nu transplantable tumor line was established. This line had the following biological properties. (1) Eighteen serial passages were carried out in 41 months. (2) Morphological changes of the grafts occurred in nude mice with serial passages: During the first 6 passages, histiological picture was consistent with the common type of MFH similar to that of the original tumor, then after the 7th passage, the myxoid type coexisted with the common type, and finally the myxoid type occupied the entire grafts to form large cysts. (3) The common type grafts grew more rapidly than the myxoid type grafts. (4) Granulocytosis (neutrophilia) was observed in mice bearing the common type tumor, but not in mice bearing the myxoid type tumor. PMID- 2562090 TI - [Establishment and characterization of TH-1 cell line from a hepatoblastoma]. PMID- 2562091 TI - [Establishment of a human hepatoblastoma (immature type) cell line OHR]. PMID- 2562092 TI - [Human hepatoblastoma cells (OHR) in collagen gell matrix]. PMID- 2562093 TI - [Clinical significance of the technetium-99m/thallium-201 overlap in acute myocardial infarction]. AB - The region of overlap of thallium-201 (Tl) and technetium-99m pyrophosphate (Tc) was evaluated as a scintigraphic prognosticator of future necrosis. Serial time courses of myocardial perfusion according to Tl and left ventricular wall motion evaluated by two-dimensional echocardiography (2D echo) were used in 22 patients. In all, dual energy emission computed tomography (dual-SPECT) showed the Tl/Tc overlap on identical slices on the third post-infarction day. According to the results of dual-SPECT, the patients were categorized in three groups: nine with large Tl/Tc overlap (group A); five with small Tc accumulation and small Tl/Tc overlap (group B); and eight with large Tl defect and Tc accumulation, which are concordant with each other (group C). Tl-201 SPECT and 2D echo were attempted serially on the 1st and 2nd days, the 7th-10th days and the 28th-30th days. To estimate infarct size with Tl-201 SPECT, we measured pixel counts of eight short axis images with the 40% cut-off level and computed "% defect". To evaluate the viability of the myocardium, "% Tl uptake" was computed from the ROIs both in the centers of the infarct areas and their border zones. 2D echoes of the left ventricular short axis at the chordae tendineae level were recorded to identify the time course of percent fractional area change (% FAC) of the ischemic left ventricular wall. The scintigraphic results were compared with the serial changes of regional ejection fraction in the areas of infarcts and ischemic lesions. The % defect remained unchanged in group C (29.2 +/- 11.5----25.7 +/- 8.3%); whereas those of groups A and B decreased significantly (21.2 +/- 11.3----9.9 +/- 6.3%, 13.8 +/- 2.6----5.4 +/- 2.9%, respectively). In groups A and B, % FAC improved significantly in the centers of the infarct areas and the border zones, but not in group C. Exercise-induced ischemia determined by redistribution of Tl at the chronic phase was observed more frequently in groups A and B than in group C. These findings indicated that more myocardium can be saved from necrosis in group A than in group C. In conclusion, it is suggested that there is considerable viable myocardium in patients with large Tl/Tc overlap on dual-SPECT. PMID- 2562094 TI - pH adjustment schedule for the amide local anesthetics. AB - Several studies have indicated that the addition of sodium bicarbonate to solutions of local anesthetics to raise the pH closer to the pKa shortens the latency, increases the intensity, and prolongs the duration of the resultant neural blockade. However, the addition of too much bicarbonate will cause precipitation, and this may result in the injection of particulate free base along with the solution. The present study was carried out to determine the maximal amount of sodium bicarbonate that can be added to each of the amide local anesthetics without the formation of a precipitate, and, thus, to construct a pH adjustment schedule to simplify the alkalinization of local anesthetics in clinical practice. PMID- 2562095 TI - The effect of pH adjustment of 0.5% bupivacaine on the latency of epidural anesthesia. AB - pH adjustment of lidocaine and 2-chloroprocaine has been reported to decrease the latency of epidural anesthesia (EA). The effect of alkalinization of bupivacaine on onset of surgical anesthesia has not been adequately studied to date. To determine what effect raising the pH of 0.5% bupivacaine has on the latency of EA in patients undergoing lower extremity surgery, we performed a randomized, double blind study. Forty patients were randomly divided into two groups. Group I patients received 15 ml of a local anesthetic (LA) solution containing 0.5% bupivacaine and 0.15 mEq of NaHCO3. Group II patients received 15 ml of a standard solution of 0.5% bupivacaine. Both solutions contained freshly added epinephrine (1:200,000). After injection of LA via Tuohy needle, sensory testing was conducted using a safety pin. The pH of the LA used for Group I was 6.96 +/- 0.01 and for Group II was 5.33 +/- 0.11. No statistically significant difference was found between the anesthetic parameters tested in each group. On this basis, we find no advantage of pH adjustment of 0.5% bupivacaine for EA. PMID- 2562096 TI - Alkalinization of epidural 0.5% bupivacaine for cesarean section. AB - Controversial results have been published in the literature concerning the efficacy of alkalinization of solutions of local anesthetics to shorten the time to onset of sensory block. Fifty-two parturients scheduled for cesarean section at term under epidural anesthesia were randomly allocated to one of four groups: group 1, 0.5% plain bupivacaine (pH = 5.38 +/- 0.05); group 2, 0.5% bupivacaine pH-adjusted with 1.4% sodium bicarbonate (pH = 6.87 +/- 0.01); group 3, 0.5% bupivacaine with 1:200,000 epinephrine (pH = 4.80 +/- 0.04); and group 4, 0.5% bupivacaine pH-adjusted with 1:200,000 epinephrine (pH = 6.68 +/- 0.01). The time to onset of the sensory block was evaluated using a nerve stimulator technique. Motor blockade was assessed using Bromage's scale. No differences in the characteristics of the onset of the sensory block were observed with epinephrine containing solutions nor with pH-adjusted local anesthetics. The maximal degree of motor blockade was not significantly different in the four groups. We conclude that alkalinization of a 0.5% bupivacaine solution is not an effective way to shorten the latency of epidural block for cesarean section. PMID- 2562097 TI - pH adjustment of local anesthetic solutions with sodium bicarbonate: laboratory evaluation of alkalinization and precipitation. AB - pH adjustment of several commonly administered local anesthetic solutions was evaluated in the laboratory. The pH achieved after the addition of sodium bicarbonate and the onset of precipitation for alkalinized solutions were recorded. Solutions of 2-chloroprocaine and lidocaine readily alkalinized to near physiological pH without precipitation. Mepivacaine solutions exhibited a tendency for delayed precipitation (18-20-minute latency for 1.5% mepivacaine) above neutral pH. Bupivacaine and etidocaine solutions precipitated after the addition of small amounts of sodium bicarbonate and could not be alkalinized to physiologic pH. Two commercially available sodium bicarbonate preparations, 4% (wt/vol) and 8.4% (wt/vol), were compared and produced similar pH changes and precipitation behavior. The data obtained for pH and time to precipitation for local anesthetic solutions alkalinized with sodium bicarbonate may be useful for practical application in the clinical setting. PMID- 2562098 TI - [Differentiation of human germ cell tumor cells]. AB - Human germ cell tumors are an excellent model for investigating the mechanism of human early embryogenesis as well as cellular differentiation. Three human EC cell lines, NCR-G 2, 3 and 4 were newly established from testicular mixed embryonal carcinomas in vitro, G3 and G4 cells were capable of somatic cell differentiation. The G3 cells demonstrated the most noticeable antigenetical changes with the administration of retinoic acid. SSEA-1 appeared on some cells whereas expression of HLA-A, B, C as well as 2H2, 2D7 and 5D4 antigens tended to be reduced in G3 cell line. 2H2, 2D7 and 5D4 antigens which we recently produced were immature human EC specific cell surface antigens, defined by mouse monoclonal antibodies, obtained immunization with G2 cells. The production of hCG, high molecular weight cytokeratin and intercellular matrices such as type IV collagen and laminin were inducible in G3 cells. Thus, G3 cells are thought to be one of the most pluripotent human EC cells. These findings clearly indicate that the EC cell lines we established provide an opportunity to study differentiation mechanism of human germ cell tumors and also human somatic cells. PMID- 2562099 TI - Starvation and germ tube formation in the exponential phase Candida albicans. AB - Two chemically defined media were developed for the induction of germ tubes in exponential phase cells of Candida albicans. One medium was N-acetyl-D glucosamine medium which is composed of L-thiazolidine-4-carboxylic acid, L proline, NaHCO3, sodium acetate, NaH2PO4 and N-acetyl-D-glucosamine. The other one was glucose medium in which N-acetyl-D-glucosamine is exchanged for glucose plus NH4Cl in N-acetyl-D-glucosamine medium. In these media, a high percentage of germ tube forming cells was obtained without a temperature shift. However, starvation of the cells in water at 37 degrees C was a necessary pretreatment to consistently obtain a high percentage of germ tube forming cells. The effect of starvation was remarkable in glucose medium, the percentages of germ tube forming cells among the normal cells and starved cells were 20 and 80, respectively. As for intracellular changes during starvation, a decrease in adenosine triphosphate concentration and an increase in adenosine 3',5'-cyclic monophosphate concentration were observed. PMID- 2562100 TI - Relationship between the pharmacological effects of benzodiazepines and their in vivo binding sites in the brain of rats. AB - To compare the in vitro pharmacological effect and in the brain distribution of benzodiazepines and phenobarbital, three group of sixty anesthetized rats (470 480 g) were administered 5 microCi of 3H-diazepam, 3H-flunitrazepam or 3H phenobarbital. The rats were decapitated 3, 10, or 40 minutes after the intravenous injection of these drugs. Radioactivity of the tissue was measured and calculated as d.p.m./g. tissue. 3H-diazepam radioactivity in the brain-stem and hypothalamus was significantly higher than in the brain cortex 3 and 10 minutes after the injection. 3H-flunitrazepam radioactivity in the brain cortex was higher than in other regions. There was no significant decrease in 3H phenobarbital brain concentration, even 40 minutes after injection. The distribution of benzodiazepine is closely related with its pharmacological effect, and this suggests that in vitro benzodiazepine binding sites are not responsible for the pharmacological action in vivo. PMID- 2562101 TI - Natural history and prognostic factors of primary hepatocellular carcinoma: study of 70 untreated patients. AB - We carried out a study of the clinical courses of 70 untreated patients with primary hepatocellular carcinoma (HCC) in order to evaluate their survival period and the prognostic factors. The median survival was two months. We evaluated ten variables of biochemical parameters and findings of hepatic scintigraphy. Among them, six variables were chosen by univariate analysis. They were serum bilirubin (cut-off value 3.0 mg/dl), alkaline phosphatase (150 IU/ml), aspartate aminotransferase (AST) (200 IU/ml), alanine aminotransferase (ALT) (50 IU/ml), reticuloendothelial (RES) dysfunction (grade 1) and multiplicity of space occupying lesions (SOL). Multivariate analysis identified three variables. The RES dysfunction and multiplicity of SOL by hepatic scintigraphy and bilirubin were considered as important prognostic factors. We found that the functional reservoir of the underlying liver and multiplicity of the origin of the tumor were the most important prognostic factors. PMID- 2562102 TI - [Effects of nifedipine and verapamil on cysteinyl leukotriene-induced contractions of the sheep uterus]. AB - Both nifedipine and verapamil, the calcium channel blockers were tested to determine their anti-leukotriene properties in pig uterine strips. These drugs in concentrations ranging from 1.10(-7)M to 1.10(-5)M significantly attenuated the constrictor effects of cysteinyl leukotrienes in pig uterus. Thus, leukotrienes C4, D4 and E4 are dependent upon Ca2+ for their myotropic activity, an action non competitively inhibited by the calcium channel blockers nifedipine and verapamil. PMID- 2562103 TI - Rotational motion of spin labelled microtubule protein. AB - Microtubule protein, isolated from porcine brain by temperature-dependent assembly-disassembly cycles, was labelled with two types of nitroxide spin labels, maleimide and isothiocyanate. Labelling was performed either in depolymerized or polymerized form of the protein. Electron paramagnetic resonance spectroscopic measurements revealed 34 ns rotational correlation time of the labels in disassembled microtubule protein which corresponds most likely to the rotational motion of the subunits. Upon polymerization, changes in the rotational dynamics of microtubule protein occurred in the temperature range of 20-30 degrees C. Polymerization process was revealed as a transition between two states, one characterizing the tubulin in its monomeric form and the other the polymeric form. In the temperature range of 20-30 degrees C, both forms (monomer polymer) of tubulin were observed. Very slow rotational motion in the millisecond time range was detected in microtubule pellet. Orientation dependence in the distribution of spin labels in macroscopically oriented microtubules was not found. PMID- 2562104 TI - The role of water in the radiation sensitization of acetone. AB - These experiments were aimed at the better understanding of the mechanism of the enhancement of radiation damage brought about by acetone in spores of Bacillus megaterium. Particularly, the intention was to examine the extent of involvement of water content in this action of acetone. The radiosensitization of acetone increased with increasing oxygen concentration and ultimately became zero at 2.1% O2 in N2. The extent of sensitization increased with rising concentration of acetone up to the first peak (50% acetone in water) under anaerobic condition. Further increase in acetone concentration resulted a maximum response seen at 90% acetone in water. To investigate the role of hydroxyl radical in the radiation sensitization caused by acetone two different types of hydroxyl radical scavengers (t-butanol and iso-propanol) were used. PMID- 2562105 TI - [Chorioadenoma destruens: presentation of 2 cases]. AB - The incidence of Gestational Trophoblastic Disease varies from country to country, however, it is notable the greater frequency for developing nations. The incidence in Mexico varies from one in 144 to 625 pregnancies. The advances that have been made in the knowledge of this disease and its classification allow an integrated diagnosis and management of gestational trophoblastic disease. This report presents two cases of Chorioadenoma Destruens in which myometrial invasion was suspected from ultrasonographic studies. One case was treated with chemotherapy; the other by histerectomy following uterine perforation. The outcome was satisfactory in both cases. PMID- 2562106 TI - [Molar pregnancy (primary or recurrent?)]. AB - A peculiar case of gestational trophoblastic disease is described. A 24 year old female with former history of three molar pregnancies, spontaneous abortion and anembryoic pregnancy was admitted because of a newly diagnosed hydatiform mole (ex novo). After uterine curettage followed by a low oral dose of methotrexate (0.5 mg/kg/day) for five days. The HCG levels determined in plasma by beta-HCG- radioinmmunoassay, became negative until four months of follow3 up. An intrauterine device was installed. She resumed HCG positivity a year later and a histerectomy was performed. A post-surgical diagnosis of invasive mole was made. Since the possibility of intercurrent pregnancy was lowered by the presence of a intrauterine device, we assumed that trophoblastic transformation into an invasive mole adopted a sort of dormant period before its resurge (resurrection) independently either from curettage of chemotherapy. PMID- 2562107 TI - Trichinella spiralis as a modulator of Shope fibroma virus. AB - After the works on the promoting effect of trichinellosis on some viral infections in rodents, many studies successively demonstrated that Trichinella spiralis confers resistance to many unrelated antigens including pathogens, such as Protozoa, Bacteria and tumour cells (B16 melanoma). Considering the above contradictory results, the present work was undertaken to study, in rabbits, T. spiralis as a modulator of Shope's fibroma virus, an oncogenic virus responsible for a benign neoplasia. Four groups of 6 rabbits each were used. The rabbits of group I, II and III were inoculated per os with 3000; 6000 and 12,000 T. spiralis larvae, respectively. The rabbits of group IV were used as controls. Thirty-five days after the inoculation, all the animals were injected at the fixed doses of 0.5 ml with dilutions (10(-1) to 10(-8] of Shope's fibroma virus given intradermally into 8 different points of the skin of each pretreated and untreated rabbits. After 9 days tumour lesions affecting the inoculating area were noticed and the DI 50/0.5 of Shope's fibroma virus was then determined for each of the 4 experimental groups. The rabbits pretreated with T. spiralis exhibited much lower virus titres than the controls, which was evidently related to a certain degree of aspecific immunity conferred by the parasite. The results indicated that T. spiralis produces, in rabbits, resistance to Shope's fibroma virus and its neoplastic effect. PMID- 2562108 TI - Patterns of improvement in resection of hepatocellular carcinoma in cirrhotic patients: results of a non drainage policy. AB - A prolonged ascitic leak through abdominal drains is a source of postoperative complications and of prolonged postoperative hospital stay after liver resection for hepatocellular carcinoma (HCC) in cirrhotic patients. Therefore we elected to abstain from routine abdominal drainage in the last 14 resections in cirrhotic livers. A significantly smaller number of patients had postoperative complications following liver resections without drainage (7%) than historical controls with abdominal drainage (59%, p less than 0.01). The number of complications related to ascites was significantly greater in patients with abdominal drainage (76%) than without (0%, p less than 0.001). Postoperative hospital stay was also significantly longer following resections with abdominal drainage (19 +/- 4 days) than in patients without (12 +/- 1 days, p less than 0.01). The long postoperative hospital stay in patients with abdominal drainage was related to ascitic discharge for a mean period of 13 +/- 10 days. No clinically significant accumulation of ascites was noted in patients without drainage. A more frequent utilization of hepatic vascular inflow occlusion did not account for the better results in the group of patients without drainage. These results suggest that routine abdominal drainage should not be used following liver resection for HCC in cirrhotic patients. This appears to be another of the technical details improving postoperative results in these patients. PMID- 2562109 TI - Effect of retinal impulse blockage on cytochrome oxidase-rich zones in the macaque striate cortex: I. Quantitative electron-microscopic (EM) analysis of neurons. AB - Our previous light-microscopic study indicates that unilateral retinal impulse blockage with tetrodotoxin (TTX) causes a reversible decrease of cytochrome oxidase (CO) in alternating rows of metabolically active zones (puffs) in the adult macaque striate cortex (Wong-Riley & Carroll, 1984b). The goal of the present study was to determine if TTX blockade adversely affects all neurons or only a subpopulation of neurons within the puffs. Three major neuronal types were identified based on mitochondrial CO activities and morphological characteristics. Type A neurons were the most prevalent, consisting of small pyramidal and nonpyramidal neurons that received only symmetrical axosomatic synapses. They had little cytoplasm and relatively low levels of CO activity, and showed the least change with TTX treatment. Type B cells were medium-to-large pyramidal neurons that received exclusively symmetrical axosomatic synapses and were moderately reactive for CO. Impulse blockage caused a decrease in mitochondrial size and packing density, but somal size remained within the control range. Type C cells were medium-sized nonpyramidal neurons contacted by both asymmetrical and symmetrical axosomatic synapses. They contained abundant darkly reactive mitochondria and presumably are metabolically the most active. This cell type suffered the greatest decrease in somal size and packing density of mitochondria, particularly the darkly reactive ones. A rare fourth cell type, type D, was a small, darkly reactive nonpyramidal variety that gave rise to somatodendritic synapses. Their low occurrence prevented statistical analysis under normal and TTX-treated conditions. These data indicate that retinal impulse blockade is most detrimental to the metabolically most active neurons in the adult primate cortical puffs. The alterations are not permanent, because the effects of TTX are fully reversible (Carroll & Wong-Riley, 1987). PMID- 2562111 TI - Immunohistochemical localization of GABAA receptors in the retina of the new world primate Saimiri sciureus. AB - A large population of amacrine cells in the retina are thought to use GABA as an inhibitory neurotransmitter in their synaptic interactions within the inner plexiform layer. However, little is known about their synaptic targets; the neurons that express the receptors for GABA have not been clearly identified. Recently, the GABAA receptor has been isolated and antibodies have been raised against it. These antibodies have proven useful for the immunocytochemical localization of the receptor, and two brief reports describing the distribution of GABAA receptor immunoreactivity in the retina have appeared (Richards et al., 1987; Mariani et al., 1987). We used a monoclonal antibody (62-3G1) against the GABAA receptor to study the retina of the New World primate Saimiri sciureus. Labeled somata were found in the inner nuclear layer (INL) and ganglion cell layer (GCL). The staining was confined to what appeared to be the cell's plasmalemma and small cytoplasmic granules. Most of the labeled neurons in the INL had small somata (5-7 microns in diameter) located at the vitreal edge of the layer. They arborized in two laminae (approximately 2 and 4) of inner plexiform layer (IPL). Ventral to the optic disc (2.5 mm) they comprised 29% of the cells present. A few of the labeled neurons appeared to be interplexiform cells or flat bipolar cells, with labeled processes that extended into both the IPL and the inner half of the outer plexiform layer. In the GCL, the labeled somata were among the largest present (13-20 microns in diameter), and 2.5 mm ventral to the optic disc they made up 15% of the cells present. Experiments in which immunoreactive somata were retrogradely labeled following the injection of fluorescent tracers into the optic tract provided a conclusive demonstration that some of the immunoreactive somata were ganglion cells. The antibody often labeled their axons in the optic fiber layer. This suggests that the GABAA receptors are transported anterogradely to the retinal terminal fields. The dendrites of the immunoreactive ganglion cells extended into the 2 laminae of labeled processes in the IPL, and their primary dendritic arbors were, at any given eccentricity, quite similar in appearance. This homogeneity suggests that they comprise a particular subset of the ganglion cells. Sections simultaneously labeled with the monoclonal antibody against the GABAA receptor and antisera against either L glutamic acid decarboxylase (GAD) or GABA revealed that the GAD/GABA was distributed much more widely in the IPL than the GABAA receptor.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2562110 TI - Effect of retinal impulse blockage on cytochrome oxidase-rich zones in the macaque striate cortex: II. Quantitative electron-microscopic (EM) analysis of neuropil. AB - Unilateral retinal impulse blockage with tetrodotoxin (TTX) induces reversible shrinkage and decreased cytochrome oxidase (CO) activity in alternate rows of supragranular, CO-rich puffs in the adult macaque striate cortex (Wong-Riley & Carroll, 1984b: Carroll & Wong-Riley, 1987). The present study extended the findings to the electron-microscopic (EM) level to determine if various neuropil profiles in control puffs exhibit heterogeneous levels of CO activity, and whether specific processes were more susceptible to intravitreal TTX than others. Within the neuropil of control puffs, 60% of the total mitochondrial population resided in dendrites, and the majority of dendritic mitochondria were highly reactive for CO. Axon terminals forming symmetrical synapses also contained darkly reactive mitochondria, whereas those forming asymmetrical synapses possessed very few and mainly lightly reactive mitochondria. Unmyelinated axon trunks, myelinated axons, and glia all exhibited low levels of CO activity. Synaptic count revealed a 3:1 ratio of asymmetrical to symmetrical synapses. Intravitreal TTX for 2-4 weeks adversely affects dendrites and symmetrical terminals much more so than other neuropil processes. There was a general decrease in darkly and moderately reactive mitochondria and an increase in lightly reactive mitochondria throughout the puffs, especially in dendrites. This indicates that afferent blockade is more detrimental to processes of higher metabolic activity. Changes also differed between central and peripheral regions of puffs, and indications of axonal and synaptic reorganization were more evident in the latter. Thus, stabilization of neuronal structure and synapses appears to be activity-dependent even in the adult. A working model of these metabolic and morphological responses to chronic TTX is proposed. PMID- 2562112 TI - Calcium/calmodulin-stimulated phosphorylation of photoreceptor proteins in Limulus. AB - Calcium (Ca2+) is thought to play a major role in the photoresponse of both vertebrates and invertebrates, but the mechanisms through which Ca2+ exerts its effects are unclear. In many systems, some effects of Ca2+ on cellular processes are thought to be mediated via activation of calcium/calmodulin protein kinase (CaCAM-PK) and the phosphorylation of specific proteins. Thus, protein substrates for CaCAM-PK in photoreceptor cells may be important in mediating the effects of Ca2+ on the photoresponse. In this study, we identify eight substrates for CaCAM PK found in both the ventral and lateral eyes of Limulus. We focus on a characterization of one of these, a 46-kD substrate. We show that its subcellular distribution in ventral photoreceptors and its isoelectric forms are identical to the 46-kD light-stimulated phosphoprotein (46A) described by Edwards et al. (1989). Furthermore, we present evidence that 46A is unique to photoreceptor cells, and that it is present throughout the cell. Based on the results of this study, and the previous study by Edwards et al. (1989), we propose that 46A is involved in mediating the effects of Ca2+ on Limulus photoreceptor cell function, and that it may be involved in dark adaptation. PMID- 2562113 TI - Mechanism and site of action of a dopamine D1 antagonist in the rabbit retina. AB - Dopamine D1 antagonists have been shown to alter drastically the spontaneous and light-evoked activity of ganglion cells in the rabbit retina (Jensen & Daw, 1984, 1986). A major target of dopaminergic neurons in mammalian retinas appears to be rod AII amacrine cells (Pourcho, 1982; Voigt & Wassle, 1987). In the present study, the following questions were addressed: (1) Do dopamine D1 antagonists alter the activity of ganglion cells through actions primarily on rod AII amacrine cells? (2) Are the effects of dopamine D1 antagonists on ganglion cells due to an inhibition of dopamine-stimulated adenylate cyclase activity? Using an isolated, superfused retinal preparation, the ability of several pharmacological agents to counteract the physiological effects of the dopamine D1 antagonist (+) SCH 23390 on rabbit ganglion cells was examined. The glycine antagonist strychnine abolished the effects of (+)-SCH 23390 on the spontaneous and light evoked activity of OFF-center ganglion cells, whereas the excitatory amino-acid antagonist kynurenic acid abolished the effects of (+)-SCH 23390 on the spontaneous and light-evoked activity of ON-center ganglion cells. The findings obtained with these antagonists can be explained in terms of the known synaptic connections of AII amacrine cells. Both 8-(4-chlorophenylthio) cyclic AMP, a membrane-permeable cAMP analog, and forskolin, an activator of adenylate cyclase, reversed the effects of (+)-SCH 23390 on the spontaneous and light-evoked activity of OFF-center ganglion cells but not ON-center ganglion cells. These findings suggest that the effects of dopamine D1 antagonists on OFF-center ganglion cells are due to an inhibition of dopamine-stimulated adenylate cyclase, with the ensuing lowering of cellular cAMP levels. The effects of dopamine D1 antagonists on ON-center ganglion cells appear, however, to be independent of intracellular cAMP levels. PMID- 2562114 TI - High p21RAS expression levels correlate with chromosome 8 rearrangements in benign human mixed salivary gland tumors. AB - The expression of RAS oncogenes in benign and malignant salivary gland tumors was studied by immunohistochemistry and by immunoblotting using monoclonal antibodies recognizing the HRAS and KRAS gene products. Twenty-eight out of 29 benign pleomorphic adenomas overexpressed p21RAS, whereas only 12 out of 18 malignant salivary gland tumors expressed the p21 protein. The expression levels were also substantially higher in the adenomas than in the malignant tumors, indicating that RAS gene activation appears to be more frequent and of greater importance for benign than for malignant salivary gland tumors. Comparisons of the p21 expression levels with the karyotypes of the pleomorphic adenomas revealed a novel correlation between high p21 expression and chromosome 8 rearrangements. As a hypothesis, it is suggested that a novel gene located on the proximal long arm of chromosome 8, most likely at band q12, is involved in the regulation of RAS gene expression. PMID- 2562115 TI - Leukemic differentiation of a mediastinal germ cell tumor. AB - The biological basis for acute leukemia associated with mediastinal germ cell tumors has remained unexplained due to lack of critical data that would illuminate the genetic relationship between the two tumors in a given patient. Here we present results of serial cytogenetic investigations on the pretreatment and posttreatment mediastinal yolk sac tumor and immature teratoma biopsies and two separate leukemic bone marrow aspirates from a patient who developed acute nonlymphocytic leukemia 11 months after the initial diagnosis of the germ cell tumor. Presence of an i(12p) in all tumor clones and trisomy 21 in one clone in the posttreatment mediastinal tumor and all leukemic clones establishes the common origin of all tumor clones and shows that in this case the leukemia was derived from the malignant germ cell clone. PMID- 2562116 TI - Characteristic karyotypic anomalies identify subtypes of malignant fibrous histiocytoma. AB - Cytogenetic analysis of short-term cultures from 25 malignant fibrous histiocytomas (MFH) revealed clonal chromosome abnormalities in 17 tumors: ten storiform-pleomorphic and seven myxoid MFH. Telomeric associations, rings, and dicentric chromosomes were present in 11 tumors and cytogenetic signs of gene amplification (homogeneously staining regions and double minute chromosomes) in four. The breakpoint distribution of the numerous structural rearrangements was nonrandom. The chromosome bands most frequently affected were 19p13 (in eight tumors; eight rearrangements gave rise to 19p+ markers, some of which looked similar, and an r(19) was found in one case), 11p11 (in seven tumors; three translocations and four deletions), 1q11 (in seven tumors; one translocation and six deletions), and 3p12 (in six tumors; all deletions). Other bands involved at least four times were 1p36, 5p15, and 20q13. Of particular clinical interest was the observation that tumors with 19p+ seemed to have a pronounced tendency to recur locally (local recurrence in five of eight tumors with 19p+ compared to one of nine in tumors without this aberration; observation period 4-16 months). PMID- 2562117 TI - Human PDGFA receptor gene maps to the same region on chromosome 4 as the KIT oncogene. AB - The gene for the human platelet-derived growth factor (PDGF) A type receptor was assigned to the proximal long arm of chromosome 4 by using in situ hybridization. Of 141 labeled metaphase cells, 74 had grains over chromosome 4, with a distinct peak at bands q11-q12. The presence of the gene on chromosome 4 was also confirmed by hybridization to chromosome specific libraries. This places the PDGFA receptor gene in the same region of chromosome 4 as the KIT oncogene, another member of the PDGF growth factor receptor subfamily. The two other members of this gene family, the PDGFB receptor and the colony stimulating factor 1 (CSF1) receptor, are closely linked on the distal half of the long arm of chromosome 5. PMID- 2562118 TI - Periocular malignant fibrous histiocytoma. AB - We present a case of periocular cutaneous malignant fibrohistiocytoma that invaded the orbit. Complete excision is recommended for atypical fibrohistiocytic tumors around the eye. Examination of specimen margins by frozen section should be undertaken with caution. It is recommended that frozen section evaluation be performed by a pathologist familiar with spindle cell neoplasm to ensure complete removal of this tumor. PMID- 2562119 TI - From E.M. microprobe analysis to NMRD studies of the lens. AB - We describe how electron microscope X-ray emission spectroscopy (XES) was the basis for recent studies of lens cytoplasm using proton nuclear magnetic resonance, NMR, relaxation methods. Although electron microscopy and NMR are very different techniques, the phosphorus distributions that were observed in whole lens using XES led to studies of proton relaxation rates in lens cytoplasm. Proton NMR is sensitive to water-protein interactions that exist in transparent lens cytoplasm. The cytoplasm is transparent because the interactions produce Fourier components in the density fluctuations of lens cytoplasm that remain small relative to the wavelength of light (1). New information on the interactions that are involved in the development and maintenance of transparent cytoplasm may be obtained using methods of proton NMRD (2). PMID- 2562120 TI - Effect of radioprotective agents on X-ray cataracts. AB - The effect of some protective agents on cataract development is briefly reviewed and new evidence is presented on the efficacy of a phosphorothioate compound (Amifostine) in inhibiting the development of X-ray-induced cataract. Morphological studies showed that at the end of 4 months, lenses from X irradiated rats which had not received any drugs showed liquefaction in the equatorial region and at the posterior pole, as well as a marked swelling of the fibers in the anterior cortex. Animals which received 1.16g/kg of WR77913 showed considerable protection against the development of radiation induced cataracts with morphological changes in the lens being less severe than in animals receiving no drugs. When animals were treated with 0.5g/kg of Amifostine (WR2721) the lenses showed much greater protection against cataract development than with WR77913. Amifostine appears to be more effective than WR77913 in inhibiting X-ray induced cataract development. PMID- 2562121 TI - Role of proteolysis in lenses: a review. AB - It has been suggested that proteases are involved in removal of damaged or obsolete proteins and/or that the activation of proteases could contribute to cataract formation. This review summarizes the properties of several recently studied lens endopeptidases including: trypsin-like protease, multicatalytic endopeptidase complex, membrane bound proteases, and calpain. Properties discussed include composition, substrate specificity, distribution, changes in activity during aging, and regulation. Additionally, properties of the lens ubiquitin conjugation system are reviewed. When possible, an attempt was made to relate these findings to whether the lens proteolytic activity was involved in clearing damaged proteins, or whether it could contribute to cataract formation. Clearing of damaged or obsolete lens proteins may involve the participation of several protease activities. Findings suggest that lens protease activities are lost at variable rates during aging, and differ in concentration between species. It was concluded that the consequence of proteolytic activity in the lens may depend closely on the compliment of proteolytic activities found. For instance, proteases causing only partial degradation of lens proteins may predominate in lenses undergoing cataract formation, while proteases assisting in the removal of partially degraded proteins are lost. The partially degraded lens proteins, as well as other denatured lens proteins, may then accumulate and lead to cataract formation. PMID- 2562122 TI - Profile of a mammalian sperm receptor gene. AB - ZP3, a glycoprotein present in the extracellular coat (zona pellucida) of the unfertilized egg, serves as a receptor for sperm and an inducer of the acrosome reaction (sperm exocytosis) during fertilization in mice. As such, ZP3 regulates the initial species-specific interactions between male and female mouse gametes. Recently, the organization and expression of the gene encoding ZP3 have been studied in some detail. These studies have led to some important findings, including the entire primary structure of the glycoprotein's polypeptide chain and the sequence of more than 11 kilobases of DNA at the ZP3 genomic locus. The latter includes the entire transcription unit for ZP3, as well as 5' and 3' flanking sequences. Of particular interest is the finding that the ZP3 gene is expressed at extremely high levels by growing oocytes and by no other cell type in the mouse. This oocyte-specific expression occurs only at a particular stage of oogenesis. Although the specific regulatory elements responsible for the highly restricted expression of ZP3 have not been identified as yet, certain organizational features of the ZP3 gene that have been described may be relevant in this connection. Further molecular analyses of ZP3 will provide additional insight into its synthesis, structure, and functions, and could have practical consequences in the context of human conception and contraception. PMID- 2562123 TI - Stimulation of protein kinase C or protein kinase A mediated signal transduction pathways shows three modes of response among serum inducible genes. AB - Activation of the signal transduction pathways mediated by protein kinase A (PKA) or protein kinase C (PKC) led to different responses of several serum inducible genes including the jun gene family, c-fos, c-myc, krox 20 and krox 24. Whereas all of these genes were stimulated by the phorbol ester TPA, a chemical activator of protein kinase C, they were differently regulated upon cAMP stimulation of the PKA dependent pathway. The proto-oncogenes jun B, c-fos, and to a lesser extent jun D were stimulated by increasing the intracellular concentration of cAMP, whereas the TPA stimulation of c-jun and c-myc was inhibited under these conditions. Krox 20 and krox 24 were insensitive to this second messenger. This study allowed us to classify these growth stimulated genes into three distinct groups distinguished by their sensitivity to elevated concentrations of intracellular cAMP. The inhibition of c-jun and c-myc expression in the presence of increased cAMP levels may be at least partially responsible for the growth inhibitory effect of this agent in Balb/c-3T3 cells. PMID- 2562124 TI - Cross-activation of the Rex proteins of HTLV-I and BLV and of the Rev protein of HIV-1 and nonreciprocal interactions with their RNA responsive elements. AB - The Rex regulatory proteins of human T-cell leukemia virus type I (HTLV-I) and bovine leukemia virus (BLV), and the Rev protein of human immunodeficiency virus type 1 (HIV-1), promote the cytoplasmic accumulation and translation of viral messenger mRNAs encoding structural proteins. Rev and Rex act through cis-acting elements on the viral RNA; these elements are named Rev- and Rex-responsive elements, or RRE and RXRE, respectively. We show that the Rex proteins of HTLV-I and BLV are interchangeable, but only the Rex protein of HTLV-I can substitute for Rev of HIV-1. Rex of HTLV-I and Rev of HIV-1 appear to act on RRE by similar mechanisms. Rev of HIV-1 does not act on the RXRE of HTLV-I or BLV. The nonreciprocal action of Rev and Rex suggests that these factors interact directly with the cis-acting RNA elements of the two viruses. PMID- 2562126 TI - [Mucinous carcinoma of the breast. An analysis of 12 cases]. AB - From 1975 to 1985 we review 12 cases of mucinous carcinoma of the breast in the "Hospital Civil de Guadalajara" and we found: one in the male sex and 11 in female. The mean age was 53 years 4 (36.3%) were nulliparas, the period between the star of symptomatology and the treatment was of 3 months to 6 years, 50% were in the upperexternal quadrant, the size was of 2 to 20 cm; 5 were pure mucinous and 7 mixed (with ductal invasive carcinoma), metastasis were presenting 9 patients and the treatment utilized in the majority of the cases was the radical mastectomy. PMID- 2562125 TI - Thyroid hormone aporeceptor represses T3-inducible promoters and blocks activity of the retinoic acid receptor. AB - In the presence of its ligand, thyroid hormone receptor (T3R) binds specifically to DNA sequences near a number of genes and induces their expression. We show that in the absence of the hormone, a T3R binding site acts in cis to decrease expression from such genes. The endogenous T3 receptors in rat pituitary cell lines are sufficient to mediate this effect, as shown by comparisons of basal levels of expression directed by transiently transfected plasmids containing the rat growth hormone promoter with wild-type or point-mutated T3 response elements (T3RE). The magnitude of the negative effect is increased by increasing the strength of the T3RE or by raising intracellular levels of T3R by appropriate transfections. T3REs exert a similar negative effect on the herpes virus thymidine kinase (TK) promoter; this effect is dependent on expression of functional T3 aporeceptor (apoT3R). Analysis of a set of T3REs of increasing strength inserted upstream of the TK promoter showed a strong correlation between the level of induced expression in the presence of hormone and the level of repressed expression in the absence of hormone. These results show that, unlike other members of the nuclear hormone receptor family, T3R binds to specific DNA sequences in the absence of hormone and exerts a negative effect on expression of linked genes. The apparent affinity of apoT3R and hormone-bound T3R for a T3RE was assessed by using varying amounts of T3R expression vector in a transfection dose response assay.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562127 TI - Insulin sensitivity of rat muscle sodium pump. AB - The aim of the present study was to examine the possible physiological responsiveness of the sodium pump to insulin in rat muscle, an effect that has never been convincingly demonstrated. The insulin stimulation of the sodium pump was estimated by two well-established parameters: ouabain binding to pieces of soleus muscle, and Na/K-ATPase activity of purified plasma membranes. For both parameters the dose dependence of the insulin effect on the sodium pump shows the characteristic bell-shaped stimulation pattern, with a maximum in the physiological hormone concentration range. This result has not been observed in previous studies where insulin concentrations two to three orders of magnitude higher were used. It can be concluded that an effect of insulin on the regulation of the Na pump in muscle might well be operating in vivo. PMID- 2562128 TI - Putative cocaine receptor in striatum is a glycoprotein with active thiol function. AB - Dopamine transporters of bovine and rat striata were identified by their specific [3H]cocaine binding and cocaine-sensitive [3H]dopamine [( 3H]DA) uptake. Both binding and uptake functions of bovine striatal transporters were potentiated by lectins. Concanavalin A (Con A) increased the velocity but did not change the affinity of the transporter for DA; however, it increased its affinity for cocaine without changing the number of binding sites. This suggests that the DA transporter is a glycoprotein and that Con A action on it produces conformational changes. Inorganic and organic mercury reagents inhibited both [3H]DA uptake and [3H]cocaine binding, though they were all more potent inhibitors of the former. n Ethylmaleimide inhibited [3H]DA uptake totally but [3H]cocaine binding only partially. Also, n-pyrene maleimide had differential effects on uptake and binding, inhibiting uptake and potentiating binding. [3H]DA uptake was not affected by mercaptoethanol up to 100 mM, whereas [3H]cocaine binding was inhibited by concentrations above 10 mM. On the other hand, both uptake and binding were fairly sensitive to dimercaprol (less than 1 mM). The effects of all these sulfhydryl reagents suggest that the DA transporter has one or more thiol group(s) important for both binding and uptake activities. The Ellman reagent and dithiopyridine were effective inhibitors of uptake and binding only at fairly high concentration (greater than 10 mM). Loss of activity after treatment with the dithio reagents may be a result of reduction of a disulfide bond, which may affect the transporter conformation. PMID- 2562129 TI - Kinetics of antiviral state in cells activated by interferons in vitro. AB - It was found that development of antiviral state in cells activated by IFN alpha and beta were quite rapid. After 6-7 h treatment IFN alpha and IFN beta protected cells completely and their effects lasted for another 9-12 h after removing them from the cultures. Then the effect declined, but some protective action remained after 48 h of incubation. IFN-gamma was different from IFN alpha or beta. It activated cells much more slowly, and could not protect cells completely before 24 h of treatment. When we compared three concentrations of IFN-alpha, the cells could not be protected completely by 5 IU/ml, and kinetics were similar with 250 and 25 IU/ml. When we added anti-IFN-alpha serum before IFN treatment, the development of the antiviral state was inhibited. The results suggested that one should adopt different ways when titrating and using different forms of IFN, and it might not be needed to use maximal tolerated doses and daily administrations, and to keep the high level of IFN for a long time. We think that these results might be useful for the clinicians considering the optimal schedule for IFN treatment. PMID- 2562130 TI - Chemotactic activity of elastin-derived peptides for human polymorphonuclear leukocytes and their effect on hydrogen peroxide and myeloperoxidase release. AB - The chemotactic activity of elastin-derived peptides (EP) for human polymorphonuclear leukocytes (PMNL) was investigated using the under agarose method. The EP were produced by digesting the bovine ligament elastin with porcine pancreatic elastase. Thus prepared digest had weak chemotactic activity for PMNL. The mean chemotactic index for all tested EP concentrations did not exceed 1.30 and was lower than that obtained with zymosan-activated serum (ZAS, n formyl-methionyl-leucyl-phenylalanine (FMLP) 2.2 +/- 0.40, 3.1 +/- 0.32, (n = 10) respectively. However, EP (50 micrograms) after injection to the mouse pleural cavity induced PMNL influx. The mean PMNL number found in this cavity was 0.09 +/ 0.03 x 10(6) for PBS and 0.18 +/- 0.03 +/- 10(6) for EP injection (p less than 0.01 n = 6). Human PMNL during 60 min incubation with EP (1 to 10 micrograms/ml) or with EP and cytochalasin B (CB 4.8 micrograms/ml) released myeloperoxidase and low amounts of hydrogen peroxide. At 1 micrograms/ml and in presence of CB elastin digest was nearly as active in myeloperoxidase release as FMLP (300 ng/ml). The values reached 17.1 +/- 2.5 and 19.7 +/- 2.1% of the total activity of whole cell lysate, respectively. The obtained results suggest that EP produced in vivo in the site of inflammation could modulate to some extent its course by enhancing PMNL influx and their activation. It seems that such mechanism of enhancement of the inflammatory response may occur in the lungs which are rich in elastin fibers. PMID- 2562131 TI - Surgery of liver tumours. PMID- 2562132 TI - Malignant fibrous histiocytoma of the pancreas. AB - A case of fibrous histiocytoma of low grade malignancy arising from the uncinate lobe of the pancreas is reported. This is an unusual site for these extremely rare tumours. Survival up to 4 years has been achieved in our patient following surgical resection. PMID- 2562133 TI - Erythrocyte sodium transport in dialyzed uremic patients. AB - To investigate the status of the Na+ concentration and ionic fluxes in red cells of human subjects with dialyzed chronic uremia, the authors measured the Na(+)-K+ pump activity as well as Na(+)-K+ cotransport (CoT), Na(+)-Li+ countertransport (CTT) and Na+ passive permeability in erythrocytes from 37 normal subjects and 23 chronic uremic patients receiving maintenance hemodialysis. The mean intracellular Na+ concentration [Na+]i value in the pre-dialytic group was significantly lower than that in control subjects (p less than .0001), but tended to recover to the normal value of [Na+]i in the post-dialytic group. The mean intracellular K+ concentration value in the post-dialytic group was significantly higher than that of the control group (p less than .001), but not significantly different from that of the pre-dialytic group. It was found that the Na(+)-K+ pump activity of erythrocytes in the pre- and post-dialytic groups markedly decreased over that of the normal control group with statistical significance (p less than .0001, respectively). The Na(+)-K+ pump activity in the post-dialytic group, however, tended to recover, but not significantly. The rate constant for ouabain-sensitive Na+ efflux in the post-dialytic group was significantly decreased over that of the normal controls (p less than .05). The authors observed a significant decrease of the Na+ CoT value (p less than .001 respectively) and rate constant for Na+ CoT (p less than .05, respectively) in the patients with pre- and post-dialytic uremia vs. that of normal subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562134 TI - A case of subcutaneous seeding of hepatocellular carcinoma after fine needle aspiration biopsy. AB - Cancer spread along the needle track following fine needle aspiration biopsy is said to be a rare complication. The authors report a case of subcutaneous implantation of hepatocellular carcinoma following ultrasono-guided fine needle aspiration biopsy. The patient, a 67-year-old Korean male was found to have a large hepatocellular carcinoma diagnosed by fine needle aspiration biopsy. Four months later, the patient felt two subcutaneous growing lumps at the previous aspiration site. The authors confirmed them histologically 11 months after aspiration. PMID- 2562135 TI - [Immune response against foot-and-mouth disease virus in cattle: effect of vaccination]. AB - Foot and Mouth Disease Virus (FMDV) is one of the most feared animal virus and vaccination still has to be used in many countries. In previous reports, using a murine model, we studied the cellular basis of immune responses against FMDV and were able to show that they are atypical. In cattle, although complete protection may be attained after only one dose of killed virus vaccine, very little is known about protection against FMDV, except for antibody responses, but practically nothing concerning the cellular basis of their immune response. Moreover, since neutralizing titers do not always correlate with protection, the potency of vaccines in controlled by viral challenge. Our aim is to study cellular immune responses against FMDV, and to search for a correlate to protection. As a first step, 55 virgin cattle from a non endemic area (Patagonia) were divided into three groups: C: non immunized controls; HS: immunized with saponine containing vaccine; and EO: with oil emulsified vaccine. After vaccination, they were carried to an endemic area (Buenos Aires), where they were challenged with live FMDV. Animals were bled immediately before and 7 days after challenge, and their white blood cells and lymphocyte subpopulations were counted. All animals showed a marked neutropenia and eosinophilia, significantly higher in HS than in EO and C groups; both parameters were significantly better in the 2nd assay. Total lymphocyte counts were normal. Lymphocyte subpopulations were assessed by immunofluorescence using monoclonal antibodies: their proportions were normal and did not change during illness in group C. Several factors could have induced the observed eosinophilia and neutropenia: parasites, stress, saponine, others.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562136 TI - [Determination of antibody levels against coxsackie B virus type 2 and type 3 in sera from blood donors using the microneutralization test]. AB - The presence of antibodies and their titers against Coxsackie B virus type 2 and type 3 (CBV-2 and CBV-3) have been investigated in 208 blood donor's sera obtained from blood Bank of Gulhane Military Medical Academy. The seropositivity against Coxsackie B virus type 2 and type 3 have been determined 59.1% (123/208) and 7.2% (15/208) respectively. These findings suggest that Coxsackie B virus type 2 infections may be present more widely than Coxsackie B virus type 3 infections in our country. PMID- 2562137 TI - Low-dose enalapril in severe chronic heart failure. AB - In a 3-month prospective, single-blind, controlled trial, 38 patients in New York Heart Association functional class III-IV were assigned to group E (n = 19): enalapril 5 mg/day in addition to the previous therapy with digitalis and diuretics, or group C (n = 19): continuation of the previous therapy. In group E, 79% of the patients improved by at least one NYHA functional class after 3 months. In group C, the functional class did not change and four patients died. The echocardiographically determined end-diastolic diameter of the left ventricle decreased in group E from 72 +/- 8 mm to 63 +/- 6 mm (p less than 0.001), and the scintigraphically determined ejection fraction of the left ventricle increased from 33 +/- 18% to 40 +/- 19% (p less than 0.002). In contrast, no significant change was found in group C. Plasma-renin activity increased in group E from 8.2 +/- 1.8 ng/ml h to 29.7 +/- 14 ng/ml h (p less than 0.001), and plasma aldosterone decreased from 47.7 +/- 7.6 ng/dl to 19.9 +/- 4.8 ng/dl (p less than 0.01). In group C no significant change occurred. Comparing the actual changes (deltas) of the NYHA functional class (p less than 0.02), end-diastolic diameter and ejection fraction of the left ventricle, plasma-renin activity, and plasma aldosterone (p less than 0.0001), a significant difference between the two groups was found. Thus, low-dose enalapril resulted in a significant improvement of the NYHA functional class in patients with severe chronic heart failure, accompanied by an improvement in left ventricular function and a decrease in secondary aldosteronism. PMID- 2562139 TI - Renal transplantation at the University of Lyon. AB - Over the last 23 years, progress in renal transplantation has dramatically decreased mortality and transplant failures, especially during the first years following the transplant. Further improvement in immunosuppression or in induction of specific unresponsiveness should, in the future, limit the incidence of late failures. More experience in transplantation has led to reduced frequency and severity of most complications. This has resulted in acceptance of patients with risk factors such as old age or infancy, poor vascular status, hyperimmunization, or requirement for several transplants (kidney + pancreas, kidney + heart, kidney + liver, etc.). Optimum organ procurement facilities will be required to meet the increased demand for kidney transplants. It is hoped that this need will be stabilized when late transplant failures will become infrequent, thus decreasing the requirements for retransplantation. PMID- 2562140 TI - Cytomegalovirus (CMV) infection complicated with renal arterial thrombosis. Rapid diagnosis of CMV infection. PMID- 2562138 TI - Differential systemic and regional hemodynamic profiles of four angiotensin-I converting-enzyme inhibitors in the rat. AB - Angiotensin-converting enzyme (ACE) inhibitors decrease blood pressure by reducing systemic vascular resistance. That the peripheral vasodilating properties of ACE inhibitors might not be homogeneously distributed in all vascular beds and might differ from one drug to another has been investigated in the normotensive rat by the pulsed Doppler technique using the active components of four different ACE inhibitors: captopril, enalapril, perindopril, and ramipril. Systemic (cardiac output and blood pressure) and regional (kidney, mesentery, hindquarter) hemodynamic responses to saline or to cumulative bolus injections (0.01-1 mg/kg) of captopril, enalaprilat, perindoprilat, or ramiprilat were continuously monitored. The effects of successive bolus injections (0.3-300 ng/kg) of angiotensinII were also investigated. The four ACE inhibitors produced an almost complete blockade of plasma angiotensin-II converting-enzyme activity (83%, 100%, 100%, and 100%, respectively), induced dose-dependent decreases in mean blood pressure, did not significantly affect cardiac output, and reduced total peripheral and mesenteric vascular resistances to the same extent. Hindlimb vascular resistance was identically decreased, but to a lower extent than total peripheral resistance by enalaprilat, perindoprilat, and ramiprilat, whereas it was increased by captopril at low doses only. Renal resistance was markedly decreased by the four drugs, and especially by captopril. The decreasing rank order for ACE-inhibitor-induced vasodilation is exactly the same (renal greater than total peripheral = mesenteric greater than hindlimb vascular resistances) as that of angiotensin-H-induced regional vasoconstriction, indicating that the vasodilator properties of ACE inhibitors are mainly due to angiotensin-II vasomotor tone suppression. None of the investigated compounds significantly affected mesenteric and hindlimb blood flows, except captopril, which lowered the latter significantly.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562141 TI - Pure red cell aplasia induced by B19 parvovirus during allogeneic bone marrow transplantation. PMID- 2562142 TI - Successful bridge to transplant under extreme conditions: implanting left ventricular assist device in intensive care unit during heart massage. PMID- 2562143 TI - Aspergillosis in a cardiac transplant patient successfully treated with itraconazole. PMID- 2562144 TI - Neuropeptide Y-containing neurons are situated predominantly outside cytochrome oxidase puffs in macaque visual cortex. AB - Layers II/III of the primary visual cortex contain a regular pattern of histochemically detectable cytochrome oxidase (CO)-rich "puffs," which differ from the interpuff regions in their thalamo-cortical and cortico-cortical connectivity, receptive-field properties, and the density of inhibitory GABA containing synaptic terminals. We used an immunocytochemical method, in combination with cytochrome oxidase histochemistry, to analyze the spatial relationship between neurons that contain neuropeptide Y (NPY) and the CO puffs. Of a total of 606 neurons, only 2.6% of the NPY-containing cells are located in the puffs, whereas the rest are situated in the interpuffs, or at the interface between puffs and interpuffs. The number of NPY-containing neurons in the puffs is substantially less than that expected in an equal volume of the interpuffs (X2 = 13.86; df = 1; P less than 0.001). These observations indicate that columns containing the puffs may differ also from those in the interpuff regions in that they contain a unique array of chemically and morphologically distinct local circuit neurons. PMID- 2562145 TI - Effect of guanine nucleotides on the dark voltage of single frog rods. AB - Single frog rods consisting of the outer segment and the ellipsoid were investigated by the whole-cell patch-clamp technique. When the recording pipette was filled with a simple intracellular medium containing potassium as the principal cation, a slow increase in dark voltage (hyperpolarization) associated with a decay of the photoresponses was observed. The hyperpolarization started at a dark voltage of -27 +/- 8 mV, followed an exponential course, and leveled out at -52 +/- 6 mV. The time constant was proportional to the access resistance of the preparations. With a pipette medium containing a 0.5 or 1.0 microM cGMP, the initial dark voltage was shifted to more positive values and the tendency of hyperpolarization was clearly attenuated. Similar results were obtained with 1 mM GTP. The effects of GDP and of ATP were less significant. In experiments with 1 mM GTP plus 1 mM ATP, the dark voltage behaved as in experiments with only GTP. The stabilizing action of GTP was amplified by EGTA so that with 1 mM GTP plus 1 mM free EGTA the dark voltage was stable at a level of -15 mV. It is concluded that the preparations lose intracellular components such as cGMP and GTP by diffusion into the recording pipette and that the losses are prevented or reduced when the pipette medium contains these nucleotides in nearly physiological concentrations. For the internal transmitter cGMP, the results suggest that its free concentration does not exceed 1 microM. PMID- 2562146 TI - Development of the nucleus isthmi in Xenopus, II: Branching patterns of contralaterally projecting isthmotectal axons during maturation of binocular maps. AB - The tectum of Xenopus frogs receives input from both eyes. The contralateral eye's projection reaches the tectum directly, via the optic nerve, and the ipsilateral eye's projection reaches the tectum indirectly, via the nucleus isthmi. Under normal conditions, the topography of the ipsilateral map relayed from the nucleus isthmi is in register with the topography of the retinotectal map from the contralateral eye. During development, the process of aligning the two maps is complicated by the dramatic changes in binocular overlap of the two eyes' visual fields which take place during late tadpole and juvenile stages. The goal of this study is to determine the branching patterns of contralaterally projecting isthmotectal axons before, during, and after the period of rapid eye migration. Isthmotectal axons were filled by anterograde transport of horseradish peroxidase (HRP) from the nucleus isthmi. The results show that crossed isthmotectal axons enter the entire extent of the tectum before binocular overlap begins to increase. Therefore, binocular overlap is not necessary for the initial isthmotectal projection to span the tectum. The density of isthmotectal branches rises dramatically at the same time that the eyes begin to shift. During the period when eye migration is most rapid, many isthmotectal axons form arbors which resemble adult arbors but which extend over greater proportions of the tectal surface. The axons appear to be directed toward appropriate mediolateral positions as they enter the tectum. Their trajectories are roughly rostocaudal, with relatively little change along the mediolateral dimension. These data, when combined with available physiological data, suggest that mediolateral order is initially established by vision-independent mechanisms but can be altered by vision-dependent mechanisms. Rostrocaudal order becomes discernable only at the time when binocular visual cues become available and appears to be established primarily on the basis of the activity of the retinotectal and isthmotectal axons. PMID- 2562147 TI - Neurotrophic and behavioral effects of occipital cortex transplants in newborn rats. AB - Cell suspensions of embryonic occipital cortex were transplanted into newborn rats with large unilateral visual cortex lesions. When the animals were adults, they were tested on a difficult visual discrimination, and subsequently their brains were analyzed for possible neurotrophic effects of the transplants on nonvisual cortical areas which normally form connections with the occipital cortex. Behaviorally, animals with lesions and transplants learn to discriminate between columns and rows of squares at a rate which is identical to normal rats while animals with lesions and no transplants are impaired. Volume and cell density measures show that the transplants also rescue neurons in cortical area 8 that would normally degenerate following the cortical lesion. No such neurotrophic effect of the transplants is found in cortical area 24 or area 17 contralateral to the lesion. In rats with lesions and no transplants, there is a significant correlation between the amount of area 8 remaining after the lesion and trials to criterion on the columns-rows discrimination, a relationship that does not exist in transplant animals because of their normal learning curve and the consistent sparing of area 8. Injections of HRP into the visual cortex contralateral to the lesion result in variable numbers of labeled cells within the transplant. However, there is no consistent relationship between the number of transplant cells which project to the opposite hemisphere and learning of the discrimination. It is suggested that the learning deficit following the lesion is largely attentional and that the sparing of cortical area 8 (which in rats may include the analog of the frontal eye fields present in the primate cortex) contributes to the sparing of function. PMID- 2562148 TI - Distribution and size of ganglion cells in the retinae of large Amazon rodents. AB - The topographical distribution of density and soma size of the retinal ganglion cells were studied in three species of hystricomorph rodents. Flat-mounted retinae were stained by the Nissl method and the ganglion cells counted on a matrix covering the whole retinae. Soma size was determined for samples at different retinal regions. The agouti, a diurnal rodent, shows a well-developed visual streak, reaching a peak density of 6250 ganglion cells/mm2. The total number of ganglion cells ranged from 477,427-548,205 in eight retinae. The ganglion-cell-size histogram of the visual streak region exhibits a marked shift towards smaller values when compared to retinal periphery. Upper and lower regions differ in both cell density and cell size. The crepuscular capybara shows a less-developed visual streak with a peak ganglion cell density of 2250/mm2. The shift towards small-sized cells in the visual streak is less marked. Total ganglion cell population is 368,840. In the nocturnal paca, the upper half of the fundus oculi includes a tapetum lucidum. The retina of this species shows the least-developed visual streak of this group, with the lowest peak ganglion cell density reaching 925/mm2. The total ganglion cell number (230,804) is also smaller than in the two other species. Soma-size spectra of this species are characterized by the presence, in the lower hemi-retina, of very large perikarya comparable in size to the cat's alpha ganglion cells. PMID- 2562149 TI - Projections of the nucleus of the optic tract to the nucleus reticularis tegmenti pontis and prepositus hypoglossi nucleus in the pigmented rat as demonstrated by anterograde and retrograde transport methods. AB - The visual pathways from the nucleus of the optic tract (NOT) to the nucleus reticularis tegmenti pontis (NRTP) and prepositus hypoglossi nucleus (ph) were studied following injections of tritiated leucine into the NOT of pigmented rats. The cell bodies of origin of the pretectal-NRTP, NRTP-ph, and pretectal-ph projections were determined using retrograde horseradish peroxidase (HRP) technique. The pretectum projects strongly to the rostral two-thirds of the central and pericentral subdivisions of the NRTP and sends a remarkably smaller projection to the ph. Both are entirely ipsilateral. The fibers destined for the ph travel with the NOT-NRTP bundle, pass through the NRTP, traverse the medial longitudinal fasciculus, and are distributed to the rostral one-half of the ph. The retrograde HRP studies confirm these pathways. The pretectal projections to the NRTP arise from neurons in the rostromedial NOT; those to the ph are located primarily in the rostral NOT although small numbers are found within the anterior, posterior, and olivary pretectal nuclei. Of major importance is the fact that the ph injections retrogradely label neurons within the NRTP and the adjacent paramedian pontine reticular formation. This NRTP-ph projection is entirely bilateral and arises from parts of both subdivisions of the nucleus targeted by NOT afferents. Both the direct NOT-ph and indirect NOT-NRTP-ph connections provide the anatomical basis for the relay of visual (optokinetic) information to the perihypoglossal complex and, presumably, by virtue of reciprocal ph-vestibular nuclear connections, to the vestibular nuclei itself. Such pathways confirm previous physiological studies in rat and, in particular, clarify the contrasting effects of electrolytic lesions of NRTP in rat which completely abolishes optokinetic nystagmus (OKN) (Cazin et al., 1980a) vs kainic acid lesions which produce only minor effects on OKN slow velocity (Hess et al., 1988). Given these differential effects, one concludes that the critical pathway for OKN passes in relation to, but is not significantly relayed by, the neurons of the NRTP or adjacent pontine tegmentum. The present studies suggest that one such fiber system is the NOT-ph bundle. How this relatively small projection compares to other possible fiber of passage systems remains to be determined electrophysiologically. PMID- 2562150 TI - Morphology of retinogeniculate axons in the macaque. AB - The size, pattern of terminal arborizations, and laminar specificity of individual retinogeniculate axons were studied in the macaque following injections of HRP into the optic tract. Axons that terminated in the magnocellular layers had significantly larger fiber diameters and wider terminal fields than those that terminated in the parvocellular layers. Terminal fields of magnocellular fibers spanned most of the width of their target layer, whereas those of parvocellular fibers were restricted to approximately one-half the width of their target layers; almost all terminal fields were oriented along lines of projection. All of the optic tract fibers that we examined terminated in only one layer of the lateral geniculate nucleus (GL), including a population of fine caliber fibers that project to the intercalated layers, and none had collateral projections outside the GL. The results suggest that each layer--magnocellular, parvocellular, and intercalated--receives projections from a morphologically distinct population of optic tract fibers. PMID- 2562151 TI - Binocular competition affects the pattern and intensity of ocular activation columns in the visual cortex of cats. AB - The effect of binocular competition on the development of ocular activation columns in areas 17 and 18 of cats was studied using the 14C-2-deoxyglucose (14C 2DG) technique to visualize the regions of cortex activated by one eye in cats reared with equal alternating monocular exposure (equal AME), unequal AME, or monocular deprivation (MD). The average size of the ocular activation columns of the eye stimulated during administration of 2DG was positively correlated with the competitive advantage during rearing. In order of increasing percentage of visual cortex activated, the eyes were (1) deprived eye of MD cats, (2) less experienced eye of unequal AME cats, (3) either eye of equal AME cats, (4) more experienced eye of unequal AME cats, and (5) experienced eye of MD cats. In area 17, the shape of the activation columns also was affected by the relative experience of the eye. The columns of the deprived eye of MD cats were widest in layer IV, where they were about the same width as those of the less experienced eye of the unequal AME cats; in other layers they were narrower, sometimes disappearing altogether. In contrast, the activation columns of the less experienced eye of the unequal AME cats were about the same width in all layers. These results suggest that when one eye is placed at a severe disadvantage and receives no patterned input, as in MD, both geniculocortical connections and intracortical connections may be disrupted, but when the disadvantage is less, as in unequal AME, only the geniculocortical connections are disrupted. Binocular competition also affected the intensity of activation within columns in area 17. We used video densitometry to determine ratios of the amount of label in cortical and thalamic structures. Both the ratio of label in area 17 to that in the lateral geniculate nucleus (LGN) and the ratio of label in the binocular segment of area 17 to that in the monocular segment were significantly less for the deprived eye of MD cats than for any other group. These results suggest that even within the smaller activation columns, deprived geniculocortical afferents are relatively ineffective at driving cortical cells. This finding is consistent with earlier reports that the synapses from the deprived pathway are both morphologically abnormal and reduced in number. The cortical labeling for the less experienced eye of the unequal AME cats and the experienced eye of the MD cats were also significantly less than that in equal AME cats.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2562152 TI - The amount of information transmitted about contrast by neurones in the cat's visual cortex. AB - The responses of neurones in the cat's visual cortex are very variable in amplitude. Thus, although the average response amplitude of a single neurone depends closely upon the contrast of a sinusoidal grating, the instantaneous amplitude of the response can convey little information about the grating's contrast. This paper shows that a typical cortical neurone can convey less than one bit of information about contrast in 0.5 s. The amount of information that a neurone can convey is closely correlated with the neurone's responsivity. PMID- 2562153 TI - Contractility effects of local anesthetics in the presence of sodium channel blockade. AB - The effects of lidocaine (25, 50 and 100 micrograms/ml) and bupivacaine (5, 10, and 20 micrograms/ml) on amplitude, velocity, and contraction frequency were studied in monolayer cultures of spontaneously beating chick embryo ventricular cells. The physiologic parameters of contraction frequency, amplitude, and velocity of shortening were measured with an optical-video system. Studies were also carried out in the presence of 1 microM tetrodotoxin (TTX) to isolate effects caused by TTX-sensitive ion channel blockade from other possible mechanisms of action of local anesthetics. Lidocaine and bupivacaine produced concentration-dependent, reversible decrements in contraction frequency, amplitude, and velocity of contraction. Bupivacaine demonstrated a 5-fold higher potency for depression of contractile properties than lidocaine. At a high concentration, bupivacaine (20 micrograms/ml) produced significantly more depression in all three measured parameters than did lidocaine (100 micrograms/ml). In the presence of predominantly Na+ channel blockade by TTX, bupivacaine (10 micrograms/ml) produced further decreases in amplitude and velocity of shortening. In the presence of TTX, lidocaine (50 micrograms/ml) produced a further decrease in amplitude of contraction, but no significant change in contraction frequency or velocity of contraction. It is well known that local anesthetics block Na+ channels of excitable membranes. The authors observations suggest that both lidocaine and bupivacaine have at least one locus of action at a site other than a TTX blockade site. PMID- 2562154 TI - The nature of the immune response (Ir) gene defect for pigeon cytochrome c in [B10.A(4R) x B10.PL]F1 mice. A comparison between thymic selection and antigen presentation. AB - [B10.A(4R) x B10.PL]F1 mice are low responders to pigeon cytochrome c, while [B10.A(2R) x B10.PL]F1 and B10.A mice are high responders. The in vivo site at which the different allomorphs of the E alpha Ia molecule exert their Ir gene effect on the immune response to pigeon cytochrome c was examined by creating two different sets of radiation-induced bone marrow chimeras. [B10.A(4R) x B10.PL]F1(b.m.)----B10.A(irr.) chimeras, which possess antigen-presenting cells (APC) of the low responder, but whose T cells are educated in a high responder environment, were found to be low responders to pigeon cytochrome c. In contrast, B10.A(b.m.)----[B10.A(4R) x B10.PL]F1(irr.) chimeras, which possess APC of the high responder type, but whose T cells are educated in a low responder environment, responded to pigeon cytochrome c. Addition of B10.A APC to the first type of chimera, both prior to antigen priming and at the time of the secondary challenge in vitro, converted 50% of the animals to responders. Furthermore, [B10.A(4R) x B10.PL]F1 mice responded to pigeon cytochrome c if they were primed with a 10-fold greater antigen dose and restimulated in vitro in the presence of B10.A APC. These results suggest that the primary site of the Ir gene defect in this system is at the level of antigen presentation and not in the T cell repertoire. PMID- 2562155 TI - T cell receptor DJ but not VDJ rearrangement within a recombination substrate introduced into a pre-B cell line. AB - To elucidate mechanisms that regulate ordered and tissue-specific assembly of Ig and TCR variable region gene segments, we have introduced a recombination substrate comprised of germline TCR beta V, D, and J gene segments into an Abelson murine leukemia virus-transformed pre-B cell line that actively rearranges endogenous Ig H chain variable region gene segments but does not rearrange endogenous light chain or TCR variable region gene segments. We find that these cells efficiently join D beta segments to J beta segments within the mini-locus, but that they do not make any detectable site-specific rearrangements of the introduced V beta segment even though it is closely linked in the same construct to the D beta. These findings suggest that factors necessary for V beta to (D beta)J beta joining may be absent in these pre-B cells and also imply that the order in which TCR V beta, D beta, and J beta segments are rearranged can be influenced by factors other than the 12/23 recombination rule. Furthermore, in agreement with the an accessibility model of VDJ recombinase control, the D beta region of the construct was found to be relatively more sensitive to DNAase I digestion in isolated nuclei when compared to the unrearranged V beta region. PMID- 2562157 TI - Specific inhibition of class II MHC gene expression by anti-sense RNA. AB - We have established an anti-sense RNA system which is capable of regulating expression of the class II (Ia) molecule coded for by the major histocompatibility complex in cultured mouse cells. Various areas of the I-A beta chain gene were subcloned in an anti-sense orientation to the 3' of the dihydrofolate reductase (DHFR) cDNA under the control of the human metallothionein IIa gene promoter. These anti-sense DNA constructs were transfected into M12.4 cells, a BALB/c B lymphoma cell line which expresses both I-A and I-E molecules on the cell surface. I-A expression of selected clones transfected with anti-sense DNA encompassing the 5' untranslated region (UT) (100 or 310 bp) including the translation start site or the poly(A) addition signalling sequence in the 3' UT (250 bp) of the I-A beta chain gene were specifically reduced to less than 5% of the control M12.4 cell surface I-A expression. These clones had normal levels of I-E expression. However, transfection of the anti-sense DNA to the beta 1 domain (510 bp) including the splicing donor and acceptor sequences did not affect the expression of I-A molecules. The same antisense DNA constructs (100 bp of the 5' UT or 250 bp of the 3' UT) without the DHFR cDNA (710 bp) did not down-regulate the expression of I-A molecules, indicating that either the physical length of the anti-sense RNA or specific DHFR cDNA sequences are also important.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562156 TI - Development of autoimmune insulitis is prevented in E alpha d but not in A beta k NOD transgenic mice. AB - Two lines of E alpha d-expressing NOD mice were established by continuously backcrossing [E alpha d B6 transgenic mice x NOD] F1 to parental NOD or directly microinjecting the E alpha d gene into fertilized NOD eggs. Similarly, A beta k expressing transgenic NOD mice were produced. Subsequent histological examination of pancreatic tissues revealed that autoimmune insulitis was prevented in E alpha d backcross and transgenic mice but not in A beta k transgenic mice. PMID- 2562158 TI - Violation of allelic exclusion of the T cell receptor beta genes in a helper T cell clone. AB - The 4-hydroxy-3-nitrophenylacetyl coupled to chicken gamma globulin specific and I-Ab restricted helper T cell clone IH4 carries a V beta 8.1-D beta 2.1-J beta 2.3-rearrangement on one and a V beta 16-D beta 2.1-J beta 2.5-rearrangement on the other chromosome. Both rearranged beta genes are transcribed and the products of both genes are expressed on the cell surface. This result implicates the absence of allelic exclusion of the T cell receptor beta genes in this T cell clone. The finding of a pseudoleader 5' to the functional leader exon of the V beta 16 gene suggests that this gene is not efficiently translated and that the amount of V beta 16 chains synthesized is not sufficient to activate the inhibitory mechanism preventing a second V beta to D beta J beta rearrangement. PMID- 2562159 TI - Post-transcriptional allelic exclusion of two functionally rearranged T cell receptor alpha genes. AB - We cloned and sequenced T cell receptor (TCR) alpha and beta chain cDNA from a lambda gt10 library obtained from a murine I-Ak autoreactive helper T cell clone MS202. Two types of cDNA clones for the alpha chain and one for the beta chain were obtained. The two alpha chain transcripts used two different V alpha genes: V alpha 4, joined to J alpha 11.2; and V alpha 5, J alpha TA13. The four V alpha 4 cDNA clones obtained did not have a complete sequences, lacking the leader portion. The V alpha 4 genomic gene segment of MS202 was revealed to contain two exons corresponding to the V alpha 4.MD13 cDNA sequence, and the potential RNA splicing signals between the two exons were intact. Both of the alpha chain cDNA clones showed in-frame rearrangements. Immunoprecipitation of 125I-surface labeled lysate of MS202 with anti-TCR antiserum and subsequent electrophonetic analyses indicated that only one of the alpha chain polypeptides was expressed on the cell surface. Thus, allelic exclusion of the alpha chain in MS202 is achieved by post-transcriptional regulation rather than rearrangements. PMID- 2562160 TI - Bronchiolo-alveolar adenocarcinoma in a horse. AB - A bronchiolo-alveolar adenocarcinoma was diagnosed in the lungs of a horse which was euthanased after protracted respiratory disease and radiological evidence of pulmonary neoplasia. Multifocal, large, firm neoplasms occurred throughout both lungs. Neoplastic lesions were not found elsewhere. Histologically the bronchiolar and alveolar architecture was retained. The cuboidal cells lining neoplastic alveoli had very vacuolated cytoplasm, while some were ciliated. Electron microscopy identified the cells as Type II pneumocytes. Numerous distended myelinoid bodies in the tumour cells accounted for the vacuolated appearance seen by light microscopy. Special stains for fat, mucin, mucopolysaccharides and glycogen failed to elucidate the nature of the substance in these vacuoles. PMID- 2562161 TI - Plasmid-aided insertion of MMTV-LTR and ras DNAs to NIH 3T3 fibroblast cells makes them responsive to 2,3,7,8-TCDD causing overexpression of p21ras and down regulation of EGF receptor. AB - TCDD administered to NIH 3T3 fibroblast cells transfected with a plasmid containing MMTV-LTR and mouse ras DNAs caused an increased level of p21ras protein and down-regulation of EGF receptor. This effect occurred only in the cells with introduced N-ras or Ha-ras under transcriptional control of glucocorticoid-sensitive MMTV-LTR but not ones without these DNAs. The MMTV-LTR ras-incorporated cells treated with either dexamethasone or TCDD grew in soft agar to form colonies (anchorage independent growth), while nontreated cells did not, indicating profound cellular changes due to activation of N-ras by these two agents. PMID- 2562162 TI - [Biological consequences of occupational exposure to elementary mercury in light of hematologic tests]. AB - Principal haematological tests in peripheral blood, non-specific phosphatases and myeloperoxidase in granulocytes were determined in a group of 89 males occupationally exposed to the elementary mercury during manufacture of chlorine. Examined males were employed for 2-26 years. Blood mercury levels did not exceed 50 micrograms per l-1. The obtained results indicate that low concentrations of the elementary mercury in the environment do not produce significant abnormalities in both quantitative and qualitative picture of peripheral blood but markedly inhibit neutrophil non-specific phosphatases activity. PMID- 2562163 TI - [Cytochemical analysis of selected enzymes in granulocytes from the peripheral blood of coke plant employees]. AB - Cytochemical reactions detecting catalase and myeloperoxidase in the peripheral blood neutrophils were carried out in coke plants employees. Intensity of the cytochemical reactions were expressed as total score and compared with the values obtained in corresponding control group. An increase in both enzymes activity, especially catalase, was seen. It is assumed that the in enzymatic activity may be related to iron-porphyrins being prosthetic group of these enzymes. PMID- 2562164 TI - [A case of infectious mononucleosis in a 2-month-old infant]. PMID- 2562165 TI - [Hypocortisolemia with neither signs nor symptoms of cortisol deficiency: research on transcortin characteristics]. AB - Low plasma levels of cortisol, as a result of primary or secondary adrenal hypofunction, is usually associated to findings of hypocortisolism. Low plasma cortisol levels without clinical manifestations have been observed in patients in which cortisol binding capacity (CBG) is altered. In this paper we report a clinically normal patient, with very low plasma cortisol levels. We investigated a possible CBG defect or the presence of a CBG binding substance as an explanation of the clinical findings. We found a normal CBG and no competing substance. Therefore, the presence of low cortisol levels suggests the presence in this patient of a receptor with increased sensitivity to cortisol, or another molecule that can be recognized as glucocorticoid by the cortisol receptor. PMID- 2562166 TI - Epinephrine-induced toxicity of human trabecular cells in vitro. AB - To investigate the possible side effects of epinephrine on the aqueous outflow pathway, we studied the response of human trabecular cells in vitro exposed daily to the drug at various concentrations. Epinephrine at 10(-6) M caused abnormal cytokinesis and cell retraction, inhibited mitosis and phagocytosis, and induced a 4- to 5-fold increase in cAMP. After 7-10 days of exposure, notable degenerative changes were observed in the trabecular cells. Within certain limits, these effects were reversible when the drug was withdrawn. Exposure to epinephrine at 10(-5) M caused cell death within 96 hours. When a 10(-7) M concentration of epinephrine was administered for up to 10 days, no degenerative changes were seen, but mitotic activity was reduced somewhat. Pretreatment with timolol reduced, but did not completely eliminate, the cytotoxic effects of epinephrine. Cells with loose contact to the substrate were affected most. The morphologic changes and the observed loss of intracellular actin filaments indicate that the deleterious effects of epinephrine are probably mediated through damage to cellular contractile proteins. Although the precise cytotoxic action of epinephrine in vivo remains to be established, our results for primary cell cultures indicate a toxic action of this drug at concentrations of 10(-6) M or higher on human trabecular cells and suggest that, for prolonged use, the maximal dose of this drug at target sites should be in the region of 10(-7) M. PMID- 2562167 TI - Cataractogenicity and bioactivation of naphthalene derivatives in lens culture and in vivo. AB - The cataractogenicity of naphthalene derivatives was investigated in a lens culture system that included the lens with an intact capsule and epithelium. The in vivo cataractogenicity of naphthalene, 1000 or 2000 mg/kg ip, also was evaluated in New Zealand white and Chinchilla pigmented rabbits. A dose-related brunescence was observed in lenses incubated with 1,4-naphthoquinone in concentrations from 31.6 to 316 microM. With 316 microM naphthoquinone, lenses were totally opaque within 24 hr. No lenticular opacities were observed with 1 naphthol or 2-naphthol in incubations lasting up to 96 hr. The bioactivation of naphthalene derivatives to reactive free radical intermediates by lenses in organ culture was investigated by electron spin resonance spectrometry (ESR) using the spin trap alpha-phenyl-N-t-butylnitrone (PBN). Lenses were incubated with 316 microM naphthoquinone and 100 mM PBN for 0.25, 4 or 7 hr. A spin trapped radical product with unresolved peaks was observed with 0.25 and 7 hr incubation. No radicals were detected in the 4 hr incubation, nor in control cultures lacking either the lens, naphthoquinone or PBN. In the in vivo studies, naphthalene was cataractogenic in both albino and pigmented rabbits. The in vitro results indicate that naphthoquinone can be bioactivated by rabbit lens to a reactive free radical intermediate, which may contribute to cataractogenicity. PMID- 2562168 TI - Isolated, perfused bovine eye a model for acute retinal toxicity screening. AB - Excorporal bovine eye has been resuscitated and sustained with an aim of its development as an alternative to whole animal in evaluating acute retinal toxicity of xenobiotics. As indicated by the stable ERG response, such preparation is functionally reactive to photic stimuli over a span of 6-12 h. Moreover, after experienced a 20 min hypoxia the reperfused eyes recovered fully. This functionally responsive preparation was used to evaluate the influence of a highly purified bacterial collagenase (Nucleolysin) on the integrity of vitreoretinal junction. Nucleolysin at concentrations between 120-600 U/ml was injected into the posterior chamber for 15 min. Retinal surfaces were irrigated and eyes were perfusion fixed with glutaraldehyde. Ultrastructural analysis indicated that thinning of the internal limiting membrane occurred when enzyme exceeded 480 U. At 600 U concentration, the principle subcellular change involved the end-foot region of the Muller cells. These studies suggest that the isolated, perfused bovine eye is a suitable model for evaluating acute retinal toxicity of xenobiotics. PMID- 2562169 TI - Oxygen free radicals and corneal endothelium. AB - We have demonstrated that corneal endothelial cells are susceptible to damage from hydrogen peroxide. Hydrogen peroxide perfusion of corneal endothelium results in physiologic and anatomic disruption of corneal endothelial cells with resulting swelling of the corneal stroma. Enzymatically generated superoxide anion produced endothelial damage which could not be blocked with superoxide dismutase (which removes superoxide anion from the system), but could it be blocked with catalase which removes hydrogen peroxide from the system. This indicated that the superoxide anion was not the toxic agent, but rather hydrogen peroxide, its dismutation product. Enzymatically generated oxygen free radical in the anterior chamber of the rabbit eye resulted in an increased iris vascular permeability which persisted for up to 24 hours following anterior chamber injection of hypoxanthine and xanthine oxidase. Young animals were better able to withstand oxidative stress to the eye then were older animals. The physiologic effects of oxidative stress in young animals can be made comparable to that in the older animals by depletion of catalase with 3-aminotriazole. PMID- 2562170 TI - Metabolic differentiation of bloodstream forms of Trypanosoma brucei brucei into procyclic forms in hemin-depleted medium and in the presence of respiratory inhibitors. AB - Bloodforms of Trypanosoma brucei brucei (STIB 247) differentiated in vitro into procyclic forms as described in the accompanying paper (Markos et al. 1989). The importance of the respiratory chain for the process was tested by the inhibition of its development (omission of hemin from the medium) or function (respiratory inhibitors). In the absence of hemin, all enzyme markers of the procyclic state, except for hemoproteins, developed to 50-70 per cent of control values. The presence of hemin is therefore not essential for the onset of differentiation, although the process cannot be completed under hemin limitation. Addition of 1 mumole.dm-3 KCN, 10 mumole.dm-3 antimycin A, or 100 mumole.dm-3 salicyl hydroxamate (SHAM) did not block the differentiation, although it proceeded at a slower rate. The development of the inner mitochondrial membrane markers- succinate: cytochrome c reductase, and NADH: cytochrome c reductase--was strongly inhibited by KCN or antimycin. None of these inhibitors had a significant effect on the activity of procyclic state marker--glycosomal malate dehydrogenase. PMID- 2562171 TI - Results of arbovirological examination of birds of the family Hirundinidae in Czechoslovakia. AB - Migratory birds (swallow, Hirundo rustica; sand martin, Riparia riparia; house martin, Delichon urbica) caught in southern Moravia (Czechoslovakia) in 1984-87 were examined for arbovirus infections. Isolation experiments were carried out using blood samples of 183 birds (52 swallows, 107 sand martins, and 24 house martins). The results were negative. Serological examinations of 136 birds (36 swallows, 86 sand martins, and 14 house martins) were made by haemagglutination inhibition test (HIT) using 6 arboviral antigens of the genera Alphavirus (Sindbis--SIN) and Flavivirus (tick-borne encephalitis--TBE, West Nile--WN) and of the family Bunyaviridae (Tahyna--TAH, Calovo, CVO, and Bhanja--BHA). Antibodies against all of the tested viruses were detected at different rates: SIN 2.9%, TBE 1.5%, WN 1.5%, TAH 4.4%, CVO 1.5%, and BHA 2.2%. The titres ranged from 1.20 to 1.80. PMID- 2562172 TI - [The role of beta-endorphin in the pathogenesis of essential arterial hypertension and obesity]. PMID- 2562173 TI - [Bolande's tumor: significance of its early diagnosis and treatment]. AB - Four patients diagnosed of having a tumor of Bolande and treated at our institution in the past ten years are presented. In two of them, early diagnosis was aided by prenatal echographic techniques; the other two diagnoses were suspected ar three and ten days of age because of an abdominal mass. The four of them underwent a complete study which included: a plain abdominal film, an ultrasound scan and an intravenous urogram; the last three patients were also surveyed by a CT scan of the abdomen. Surgery was undertaken early on all the patients, the time of operation being in every instance within the first month of life. A laparotomy was performed and, once the contralateral kidney had been carefully examined, a nephroureterectomy was implemented, having been able to preserve in all the cases the suprarenal gland in view of the benign nature of the tumor. A histologic study confirmed the diagnosis and revealed the complete delimitation of the tumor by the renal capsule. The authors underscore the importance of this tumor's suspicion in the face of any solid renal mass detected by prenatal echography or in the neonatal period, since early diagnosis and treatment are imperative in order to prevent the ensuing complications in the rare and unfortunate cases that show a malignant tumoral trend. PMID- 2562174 TI - Genetic aspects of oncogenesis by murine leukemia viruses in wild mice. PMID- 2562175 TI - Fluconazole therapy for fungal peritonitis in continuous ambulatory peritoneal dialysis (CAPD): a case report. AB - Fluconazole proved effective in treating fungal peritonitis caused by Trichosporon cutaneum. Fluconazole seems to offer several advantages over other antifungal drugs in the treatment of fungal peritonitis in continuous ambulatory peritoneal dialysis (CAPD) patients. PMID- 2562176 TI - Isolation and characterization of new mammalian kinase genes by cross hybridization with a tyrosine kinase probe. AB - We isolated two novel mammalian kinase genes by weak cross-hybridization with v ros oncogene. (1) A cDNA of about 7.7 kb obtained from a human placenta cDNA library carried a 4.2 kb open reading frame, and the predicted amino acid sequence of 1338 residues contained extracellular, transmembrane and tyrosine kinase domains. The overall structure including cysteine motifs in its extracellular domain and a long peptide insertion in its tyrosine kinase domain indicates that this new gene is closely related to the fms family. Consequently, the gene was designated as flt(fms-like tyrosine kinase) gene. The expression of the flt gene was examined in normal and transformed cells, and it was mapped to human chromosome 13q12-13. (2) Another new gene was found to be expressed almost exclusively in testis. The cDNA sequence analysis revealed that the predicted product carried protein kinase consensus motifs in its amino-terminal region. Comparison of the deduced amino acid sequence of this gene in the kinase domain with those of other protein kinase genes will be discussed. PMID- 2562177 TI - The oncogenicity of jun. AB - The oncogenic transforming potential of the jun oncogene was investigated with viral constructs that contain various terminal deletions of v-jun, c-jun and recombinants between the viral and cellular genes. Cellular jun can induce transformation of chicken embryo fibroblasts with a low efficiency. High efficiency transformation is dependent on alterations in the major transactivator domain. A jun deletion that lacks the transactivator domain is non-transforming. These observations are compatible with the conclusion that transcriptional regulation plays an important part in jun oncogenesis. In cells transformed by viral jun and expressing high levels of viral jun expression of the cellular jun is not constitutively upregulated. Cellular jun may therefore not contribute to transformation in these cells. PMID- 2562178 TI - Forward and reverse changes in Ig/myc translocation carrying tumors. AB - Translocation of the c-myc protooncogene to an immunoglobulin locus is a rate limiting step in the genesis of three B cell derived tumors: Burkitt lymphoma (BL) in humans, mouse plasmacytoma (MPC), and rat immunocytoma (RIC). The translocation appears as a rate-limiting step in the genesis of all three tumors. Its consequences have been best analyzed in BL. They involve a non-immunological and an immunological component. The former acts by preventing the B cell from leaving the cycling compartment and entering the resting stage when programmed to do so. The latter acts by the down-regulation of certain human leucocyte antigen (HLA) class I polymorphic specificities, adhesion molecules and Epstein-Barr virus (EBV) encoded proteins. Together, they contribute to the escape of the BL cell from the host immune response. We have also described a non-clonogenic, non tumorigenic "revertant", subline among five EBV-convertants of an originally highly tumorigenic, EBV-negative BL line. The other four convertants have remained highly tumorigenic. Suppression of tumorigenicity is associated with a switch to a lymphoblastoid line (LCL)-like phenotype, accompanied by the appearance of several activation markers. It is suggested that the LCL-type immunoblast comes under the influence of host feedback that normally contribute to the constancy of the B cell pool. PMID- 2562179 TI - Wilms tumour: a developmental anomaly. AB - Wilms tumour (WT) is a developmental anomaly of the kidney which results from loss of function of at least one so called tumour suppressor gene on chromosome 11. The position of the gene at chromosome 11p13 is known through the association of WT with aniridia (lack of an iris), mental retardation and genitourinary abnormalities in the WAGR syndrome. Here we discuss the high resolution mapping studies to locate the position of the gene and conclude that the gonadal abnormalities in WAGR patients may be due to a defect in the WT gene itself. In support of this role in genitourinary development we show that a candidate WT gene is expressed in specific regions of the developing kidney and in fetal and embryonic gonads. PMID- 2562180 TI - Phosphorylation of the tumor suppressor gene RB protein by M-phase specific histone H1 kinase. AB - We have noted the presence of the consensus amino acid sequence for phosphorylation by M-phase specific histone H1 kinase in six sites of the tumor suppressor gene retinoblastoma (RB) protein and determined whether or not RB protein is, in fact, phosphorylated by this kinase. Highly purified enzyme was used for this purpose. Human cell extracts immunoprecipitated with anti-RB antiserum as well as RB proteins expressed in Escherichia coli cells were shown to be phosphorylated by this kinase in vitro. Synthetic peptides for the six expected sites were also phosphorylated. These results suggest the possibility that the function of RB protein is regulated by CDC2 kinase. We also noted the presence of putative phosphorylation sites by H1 kinase in a homologous region between the RB gene and L1 family repetitive sequences. PMID- 2562181 TI - Role of protein phosphatases in malignant transformation. AB - Many oncogene products are protein kinases and signals are transduced via phosphorylation of proteins. Similarly, protein-dephosphorylation may play a critical role in malignant cell transformation. We have cloned two catalytic subunits of ser/thr protein phosphatase (PP) type 2A, PP2A alpha, and PP2A beta, from a rat liver cDNA library. Both cDNAs encode peptides of 309 amino acids with a difference of only 8 amino acids between the two. All primary hepatocellular hyperplastic nodules or carcinomas, which were induced by a food carcinogen, 2 amino-3-methylimidazo[4,5-f]quinoline, showed up-regulation of expression of the mRNAs of both PP2A alpha and PP2A beta. NIH3T3 cell transformants obtained by introducing activated c-raf, ret-II or Ki-ras oncogenes also showed high levels of PP2A alpha transcripts. Okadaic acid, a non-TPA type tumor promoter, was found to be a potent inhibitor of PP1 and PP2A. Its IC50 for PP1 was much higher than that for PP2A with phosphorylase a as a substrate. When raf and ret-II transformants were cultured with okadaic acid at 8 ng/ml for 2 days, both transformants became flattened and showed strict contact inhibitions. This flat cell morphology was stable for at least one month in the presence of okadaic acid, but in its absence, the cells reverted to their original transformed shape within 7-10 days. Colony formation by raf and ret-II transformants in soft agar was inhibited dose-dependently by okadaic acid; very few colonies grew in the presence of the acid at 8 ng/ml. Okadaic acid had less effect on a transformant of the Ha-ras gene, causing only 50% inhibition of colony formation at 8 ng/ml. The role of protein phosphatases in cellular transformation by certain oncogenes is suggested. PMID- 2562182 TI - Molecular mechanisms of transformation by the human papillomaviruses. AB - A variety of studies have now indicated that the papillomaviruses associated with cervical carcinoma, HPV-16 and HPV-18, contain two genes (E6 and E7) which have transforming properties. These genes are generally expressed in cervical carcinoma cells. The HPV-16 E7 protein has recently been shown to be a multi functional protein possessing both transcriptional modulatory and cellular transforming properties similar to those described for adenovirus E1A proteins (1). E7 is able to transactivate the adenovirus E2 promoter and can cooperate with an activated ras oncogene to transform primary baby rat kidney cells. The N terminal 37 amino acids of all of the E7 proteins of the genital associated HPVs contain regions which are highly conserved and which are quite similar to portions of conserved domains 1 and 2 of adenovirus E1A. These domains in E1A are critical for cellular transformation properties and contain the amino acid sequences involved in binding the product of the retinoblastoma tumor suppressor gene (pRB). Results from a collaborative study with Ed Harlow and Nick Dyson (Cold Spring Harbor Laboratory) have shown that the E7 oncoprotein of HPV-16 can associate with the retinoblastoma gene product in vitro (2). The ability of the E7 proteins encoded by various HPVs to bind pRB has been examined using an in vitro complexing assay. E7 is not sufficient for transformation of human keratinocytes. The co-operation of the HPV-16 E6 and E7 genes has been shown to be important for transformation of these cells (3). Potential intracellular protein targets for E6 are being assessed. PMID- 2562183 TI - Host cell regulation of HPV transforming gene expression. AB - Infections with specific types of human pathogenic papillomaviruses, most notably HPV 16 and 18, appear to be necessary but not sufficient factors in the etiology of anogenital cancer. Recently their role in the induction of genital intraepithelial neoplasias became evident. The expression of specific HPV genes (E6-E7) emerges as an important prerequisite for the proliferative phenotype of cervical carcinoma cells. Increasing evidence points to the existence of a host mediated intracellular control which down-regulates these HPV genes in replicating normal cells. This control appears to be interrupted in HPV-positive carcinoma-cells, probably due to structural modifications of the respective host cell genes acquired during the period of viral DNA persistence. Factors affecting genes seem to be responsible for geographic differences in anogenital cancer incidence, since HPV infections appear to occur worldwide at similar frequency. PMID- 2562184 TI - Transforming growth factor-beta and suppression of carcinogenesis. AB - Transforming growth factor-beta (TGF-beta) plays an important role in controlling proliferation or differentiation in almost all epithelial tissues. The pathophysiology of TGF-beta during carcinogenesis is now an important area of investigation, since it appears that as the process of carcinogenesis progresses, epithelial cells often become refractory to the growth-regulatory actions of TGF beta. In this article we consider the possible cellular and molecular bases for this phenomenon, and then discuss some pharmacological approaches to enhancing the synthesis or activity of TGF-beta. These approaches may provide new modalities for prevention of carcinogenesis, if they can be applied during the early stages of the disease process, before cells become refractory. We give particular attention to tamoxifen and retinoic acid, since it has been shown that these agents, which are of known efficacy for prevention of cancer, can markedly enhance the secretion of specific isotypes of TGF-beta by several types of cells. PMID- 2562185 TI - MyoD and achaete-scute: 4-5 amino acids distinguishes myogenesis from neurogenesis. AB - The myogenic determination gene MyoD contains a 60 amino acid domain that is necessary for both sequence-specific DNA binding and myogenic conversion of transfected C3H10T1/2 mouse embryo fibroblasts. We have generated deletion, insertion, and substitution mutants to probe the structure and function of this region, both in vitro and in vivo. Our results are consistent with a previous proposal that a helix-loop-helix (HLH) motif mediates protein dimerization. A highly basic region located immediately upstream of the HLH motif is required for DNA binding, which occurs if and only if the HLH motif is capable of dimerization. All mutants of MyoD that do not bind DNA in vitro fail to activate myogenesis or expression of a co-transfected muscle-specific reporter gene in transfected C3H10T1/2 cells. Replacing either helix 1, helix 2, or the loop sequence of MyoD with the analogous sequence of the Drosophila T4 achaete-scute protein (which is required for neurogenesis) has no substantial effect on DNA binding or biological activity. However, replacing the basic region of MyoD with the analogous sequence of the kappa immunoglobulin enhancer binding protein E12, or the T4 achaete-scute protein, produces proteins still capable of specific DNA binding in vitro, yet without significant biological activity. These findings suggest that within the 13 amino acid sequence of the MyoD basic region lies a recognition code that determines muscle-specific gene expression, although specific DNA binding per se is not sufficient to activate the muscle program. PMID- 2562186 TI - Tumorigenic latency and separable stages during fibrosarcoma development in transgenic mice carrying papillomavirus genomes. AB - Transgenic mice have been established carrying the genomes of bovine papillomavirus type 1 (BPV-1), and human papillomaviruses types 5 and 18. Transcriptional dormancy is characteristic of all three viral genomes in transgenic mouse lines maintained for 2-4 years. Only BPV-1, which induces both dermal and epidermal pathology in its natural host, has been found to elicit abnormalities when carried in transgenic mice. The BPV-1 genome acts in these mice as a tissue specific oncogene, in that it elaborates the development of skin fibrosarcomas. Three abnormal stages are evident: two distinct and successive stages of a proliferative hyperplasia (a mild and an aggressive fibromatoses), and the solid tumors (fibrosarcomas). Analysis of tissue biopsies and of derivative cell cultures from each pathology confirms that this pathway is composed of separate stages. Notably, progression from hyperplasia to neoplasia is accompanied by specific cytogenetic changes, which appear necessary in addition to the actions of the BPV oncogenes. The reproducibility of the tumorigenic pathway induced by the BPV genome is providing inroads into the molecular genetic and biochemical mechanisms of tumor development. PMID- 2562187 TI - Genetic analysis of colorectal cancer. AB - Adenomatous polyposis, mainly of the colon, (APC) is a rare dominantly inherited susceptibility to colon cancer in which individuals develop hundreds of polyps mainly in their large bowel. The APC gene has been localised to chromosome 5q21 by following up a case report of an individual with an interstitial deletion on chromosome 5q who had multiple developmental abnormalities together with adenomatous polyposis. A DNA marker (D5S71) was found to be closely linked to APC in family studies and localised to 5q21 by in situ annealing. Material from further patients with deletions in this region of chromosome 5 has been used, by a combination of somatic cell hybrid and long-range DNA analysis, to identify new DNA markers close to the APC gene. These and other markers now provide the basis for genetic counselling of nearly all families with APC. These studies are being extended, together with other approaches for analysing DNA clones around the APC gene, in the search for the gene itself. Allele loss in tumour as compared to normal tissue from sporadic cases of colorectal carcinomas has clearly implicated the APC gene in at least 25 to 40% of all cases of colorectal carcinomas. Similar studies by Vogelstein and others as well as ourselves have further implicated recessive changes on chromosomes 17 and 18 in the development of colorectal carcinomas. Following the demonstration by Vogelstein of the role of p53 mutations in connection with the chromosome 17 changes, we have now shown, using monoclonal antibodies to the mutant p53 products and by other approaches, that changes in the p53 gene may occur in up to 50% or more of colorectal carcinomas. Frequent mutations of the K-ras dominant oncogene, as well as changes in the expression of human leucocyte antigen (HLA)-A, B, C determinants, are further genetic changes that appear commonly to be involved in the progression of colorectal carcinomas. The latter have important implications for T cell immune response to tumours and its manipulation for treatment and even prevention of colorectal cancer. We may soon be approaching a situation when it will become possible to identify all the genetic steps and their sequence during tumour progression, as well as their functional significance largely through the induction of inappropriate growth and the suppression of differentiation. PMID- 2562188 TI - The acidic amino-terminal region of the HIV-1 Tat protein constitutes an essential activating domain. AB - The Tat protein encoded by the human immunodeficiency virus (HIV) is an efficient activator of HIV gene expression. Many eukaryotic transcriptional activators contain a nucleic acid binding domain and a separate activating domain. These activating regions are acidic and often amphipathic. The amino terminus of the HIV-1 Tat protein is acidic with a periodicity of acidic, polar, and hydrophobic residues consistent with that of an amphipathic alpha helix. This region appears to be important for Tat function. We have analyzed the functional significance of acidic residues within the amino-terminal region of Tat by means of site-directed mutagenesis and by testing the capacity of mutant proteins to trans-activate the viral long terminal repeat (LTR) Conservative changes (acidic to acidic) were well tolerated, whereas acidic to neutral and acidic to basic changes markedly reduced Tat activity. The relative importance of each of the three acidic residues correlated with proximity to the amino terminus. Substitution of the entire domain with heterologous sequences that might form an acidic, amphipathic alpha helix partially restored activity when compared with an amino-terminal truncation mutant. In contrast to the observed importance of acidic residues, hydroxylated residues between amino acids 40 and 47 were dispensable for Tat function. These data suggest that the acidity of the amino terminal region is important for Tat function and that Tat-mediated trans-activation may be similar to that of other known activator proteins. PMID- 2562189 TI - The E7 protein of human papillomavirus type 16 is phosphorylated by casein kinase II. AB - The E7 protein of human papillomavirus type 16 (HPV16) transforms cultured cells and cooperates with the ras or fos oncogenes in the transformation of primary cells. In this study we have investigated the phosphorylation of E7. When we immunoprecipitated E7 from CaSki cells with a rabbit polyclonal antiserum to a bacterial fusion protein (trpE-E7), we found that E7 was phosphorylated at serine residues contained in five characteristic thermolysin peptides. Immunoprecipitated E7, and fusion proteins harboring the E7 protein from various HPV types, could all be specifically phosphorylated in vitro by the ubiquitous, growth factor-activated casein kinase II (CKII). Comparative peptide mapping showed that the sites of in vivo and in vitro phosphorylation are the same. CKII was shown previously to specifically phosphorylate serine or threonine residues within a cluster of acidic amino acids. The E7 protein contains such a sequence between amino acids 30 and 37. When a synthetic peptide corresponding to this region of E7 was phosphorylated by CKII in vitro, its thermolysin digestion products were the same as those in the phosphorylated E7 protein. We conclude that E7 is phosphorylated in vivo only at serines within the predicted CKII site and that CKII, or a CKII-like enzyme, participates in the reaction. Both the E1A and SV40 large T proteins contain similar CKII consensus sites proximal to the regions required for their associations with the retinoblastoma gene product (p105Rb). Thus it is conceivable that CKII phosphorylation can modulate the interaction between the transforming proteins and the retinoblastoma gene product. PMID- 2562190 TI - Serologic, virologic, and histopathologic observations of encephalomyocarditis virus infection in mummified and stillborn pigs. AB - Stillborn and mummified swine fetuses from swine farms experiencing reproductive problems were investigated for evidence of infection with encephalomyocarditis (EMC) virus by fetal serology, virus isolation, and histopathologic examination. Fetal sera or thoracic fluids of 478 abnormal fetuses collected during January through December 1987 were tested for the presence of antibody specific to EMC virus. Of 478 samples tested, 175 (36.6%) had EMC virus serum neutralizing antibody titers of 1:64 or greater. The samples positive for EMC virus antibody were obtained from 38 swine farms located in 9 states in the United States. In addition to serologic observations, tissue samples of some abnormal fetuses were examined for the presence of virus and histopathologic lesions. The EMC virus was isolated in 1 case from the fetuses of an aborted litter. The isolate was serologically identical to a reference EMC virus. Nonsuppurative encephalitis and myocarditis were observed in the fetal samples collected from 2 different herds. Thoracic fluid of 1 stillborn pig with lesions was positive for EMC virus antibody at a titer of 1:512. We suggest that a widespread reproductive problem recently experienced in several major swine-producing areas of the United States may have been caused by EMC virus infection. PMID- 2562191 TI - Demonstration of infectious bovine rhinotracheitis virus antigen in paraffin sections. AB - Nine pregnant heifers were inoculated intravenously with infectious bovine rhinotracheitis virus (IBRV) in the sixth month of pregnancy. Tissues were collected from the fetus of a heifer killed 13 days postinoculation (PI), from fetuses of 6 heifers that aborted 16-27 days PI, and from mummified fetuses of 2 heifers that aborted 53 and 85 days PI, respectively. Control tissues were obtained from the fetus of a non-inoculated heifer that was killed in the seventh month of gestation. Tissues were fixed in 10% formalin, embedded in paraffin, and examined for viral antigen by immunohistochemistry, using biotinylated second antibody and alkaline phosphatase-labeled avidin-biotin complex. Antigen was detected in at least 1 tissue from the fetus of each inoculated heifer. Positive tissues included lung, liver, spleen, kidney, adrenal, and placenta. In several fetuses, antigen was identified in tissues from which virus was not isolated in cell culture. This appeared to occur when tissues had only a few small foci of infection or when tissues were severely autolyzed. The observation of viral antigen in tissues from mummified fetuses indicates that this technique may be useful in diagnostic laboratories to detect IBRV infection in tissues that are not suitable for virus isolation or for examination by the cryostat tissue section-fluorescent antibody technique. PMID- 2562192 TI - Serologic examination of aborted ovine and bovine fetal fluids for the diagnosis of border disease, bluetongue, bovine viral diarrhea, and leptospiral infections. AB - Fetal serum from most of 994 bovine and 553 ovine aborted fetuses was tested serologically for antibodies to border disease (BD), bovine viral diarrhea (BVD), and bluetongue (BT) viruses, and to Leptospira sp., and the results were compared with the results of isolation procedures, fluorescent antibody tests (FAT), and histologic examinations of the same fetuses. Antibodies to BT virus were not found in any of the 994 bovine and 553 ovine fetuses. Antibody titers to BVD virus were present in 39 of 966 bovine fetuses tested, and BVD virus was detected in 4 of the 39. Four of 74 fetuses in which the BVD virus was detected by FAT or isolation had titers to BVD virus. Microagglutination (MAT) titers to 1 or more of 5 serovars of leptospires were present in 52 of 773 bovine fetal sera tested. Leptospires were not detected by FAT in any bovine fetuses that had leptospiral antibody titers. Leptospires were detected by FAT in 15 aborted calves, and none of these had MAT titers. Antibody titers to BD virus were present in 80 of 486 fetal lamb sera tested, and the virus was detected by FAT or isolation in 3 of the 80 fetuses. Border disease virus was detected in 14 of 486 fetal lambs tested. Twelve of the 14 were tested serologically and 3 had titers to BD virus. Leptospiral antibody titers were present in 27 of 326 ovine fetal sera tested. Leptospires were not detected in any of the 326 ovine fetuses tested by FAT. PMID- 2562193 TI - Detection of bovine herpesvirus-1-specific IgM using a capture enzyme immune assay with isotype-specific monoclonal antibodies. AB - The detection of virus-specific immunoglobulin M (IgM) antibodies in acute-phase serum samples offers the possibility of making an accurate and rapid serologic diagnosis. We have developed a solid-phase capture assay that uses murine monoclonal antibodies specific for bovine IgM to separate the whole IgM fraction of a bovine serum sample. The IgM specific for bovine herpesvirus-1 (BHV-1) is then detected by the addition of viral antigen, which in turn is detected by BHV 1-specific monoclonal antibodies conjugated to horseradish peroxidase. A BHV-1 IgM antibody response was detected during the early postinfection period (7-40 days PI). Bovine herpesvirus-1 IgM antibody was not detected in sera taken from 3 animals following dexamethasone-induced viral reactivation. This method compares favorably with viral isolation, antigen detection in the clinical samples, and paired serology in the diagnosis of BHV-1 infection at a herd level. PMID- 2562194 TI - A bacteriologic study of scabby-hip lesions from broiler chickens in Texas. AB - Broilers from commercial flocks experiencing a 10-60% incidence of scabby-hip lesions at processing were examined, and selected skin lesions were cultured. Over 70% of the lesions were associated with traumatic excoriations, particularly on the caudal dorsal convexity of the birds. Most lesions were observed on birds that were 5 weeks of age or older. From the 27 specimens cultured, Clostridium perfringens was isolated in pure culture from 4 lesions and Staphylococcus species from 10 lesions. Pure cultures of staphylococci were recovered from 4 lesions, and 2-5 different staphylococci were isolated from 6 lesions. Eight staphylococci were identified as S. sciuri, 8 as S. simulans, 2 as S. epidermidis, 2 as S. lentus, 2 as S. warneri, 1 as S. cohnii, and 1 as S. intermedius. Fifty cutaneous specimens from 10 5-week-old normal broilers were cultured. A total of 197 isolates were identified including 65 S. sciuri, 52 S. lentus, 24 S. simulans, 12 S. hyicus, 11 S. warneri, 9 S. cohnii, 9 S. gallinarium, 8 S. xylosus, and 7 S. epidermidis. PMID- 2562195 TI - Use of in situ hybridization with a biotinylated probe for the detection of bovine herpesvirus-1 in aborted fetal tissue. AB - Forty-five cases of bovine abortion were examined using in situ hybridization (ISH) with a biotinylated DNA probe specific for bovine herpesvirus-1 (BHV-1). Of the 45 cases, 16 were diagnosed as due to BHV-1, 15 were determined to be due to other causes, and 14 were of undetermined etiology. Direct comparisons between ISH and an immunoperoxidase (IP) test specific for BHV-1 were performed on formalin-fixed, paraffin-embedded tissue sections of lung, liver, kidney, spleen, thymus, and placenta; fluorescent antibody tests for BHV-1 and virus isolation were performed on fresh lung and liver. In comparison to these routine BHV-1 detection techniques, ISH had an overall sensitivity of 88.2% and a specificity of 89.3% in detecting BHV-1 in aborted fetuses. Immunoperoxidase was more sensitive than ISH with tissue sections from lung (87.5% vs. 69%), liver (92% vs. 17%), spleen, and placenta; results of the tests on tissue sections from kidney were concordant. Liver sections presented special problems in that nonspecific reactions were frequently observed with hybridization. With thymus sections, the rate of detection was higher by hybridization than by IP, but the specificity of some of these reactions could not be confirmed. PMID- 2562196 TI - Recombinant cDNA probe from bluetongue virus genome segment 10 for identification of bluetongue virus. AB - Genome segment 10 of bluetongue virus (BTV) serotype 11 UC8 strain was cloned and subsequently hybridized to viral double-stranded RNA extracted from 90 field isolates of BTV serotypes 10, 11, 13, and 17; the prototype strains of BTV 2, 10, 11, 13, and 17; the prototype strain epizootic hemorrhagic disease virus (EHDV) serotype 1; and 4 field isolates of EHDV serotype 2. The 90 field isolates were obtained from different counties in California, Louisiana, and Idaho during the years 1979, 1980, and 1981. The cloned genetic probe hybridized with all the BTV samples tested, showing different degrees of cross-hybridization at the stringency conditions used in this study. This indicated that BTV genome segment 10 has conserved nucleotide sequences among the BTV serotypes 2, 10, 11, 13, and 17. No cross-hybridization signals were detected between the cloned genome segment 10 of BTV 11 UC8 strain and the prototype strain of EHDV serotype 1 and the field isolates of serotype 2. This probe recognized a wide variety of BTV isolates. PMID- 2562197 TI - Genotypic transitions among bluetongue viral isolates from domestic ruminants in Colorado during 1981-1984. AB - Two predominant electropherotypes of bluetongue virus (BTV) serotype 11 isolates from cattle during a 1981-1984 field study in eastern Colorado were characterized. The genomes of strains isolated from the first 2 years of the study had 1 predominant electropherotype (CO81), with the exception of 1 isolate that differed only in the migration of segment 3. A second electropherotype (CO83), with differences in the migration of 4 segments, coexisted in the same region during 1983 and 1984 with strains having the CO81 RNA profile. The genomes of CO81 and CO83 were also distinguishable from those of the US prototype of BTV 11. Analysis of the polypeptides of representative strains of each electropherotype by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the proteins were very similar. The occurrence of the CO81 electropherotype was apparently the result of multiple viral infections since the positions of 7 segments had faint second bands and single-banded variants were isolated after serial plaque purifications. In addition, protein 7 of 1 of the CO81 isolates and protein 7 of the single-banded variant differed as shown by reverse phase-high performance liquid chromatography of 35S-methionine-labeled tryptic peptides. PMID- 2562198 TI - Susceptibility of a fetal tongue cell line derived from bighorn sheep to five serotypes of bluetongue virus and its potential for the isolation of viruses. AB - A cell line (BHFTE) was derived from a tongue explant of a bighorn sheep fetus (Ovis canadensis nelsoni). The cells have been maintained through 23 serial passages, and the modal number of chromosomes was calculated to be 55. Monolayer cultures were shown to be susceptible to various viruses, including bluetongue virus (BTV). Of 5 BTV serotypes (2, 10, 11, 13, and 17) tested, each produced a cytopathic effect (CPE) on initial passage at 33 C. A field isolate (serotype 10) of BTV from a black-tailed deer (Odocoileus hemionus columbianus) in its second passage in Vero-M cells also produced CPE when inoculated into BHFTE cells. Antigens of BTV were demonstrated by direct immunofluorescence in the cytoplasm of BHFTE cells inoculated with homogenates of chicken embryos injected with clinical specimens from a domestic sheep and an Arabian oryx (Oryx gazella leucoryx). A suspension of BTV-infected gnats (Culicoides spp.) produced CPE and BTV-specific fluorescence on the first passage in cells inoculated with a suspension of blood from sheep experimentally infected with BTV. Additionally, selected bovine viruses induced CPE in the cells. The cell line, which is free of mycoplasma and bovine viral diarrhea virus contamination, may be useful in diagnostic medicine and research involving the ruminant species. PMID- 2562199 TI - Association of Reoviridae particles in an enteric syndrome of poults observed in turkey flocks during 1988. AB - An enteric syndrome of turkey poults, characterized by enteritis, crop mycosis, intestinal changes (pale, thin-walled ballooning with watery contents), and rickets, occurred during 1988 in 74 turkey flocks from different farms belonging to 9 California turkey growers. The flocks ranged in size from 9,000 to 120,000 birds. Pools of intestine sections from 618 birds, representing 78 field cases, were examined. Histopathological examination of the intestines showed a mild to severe atrophy with a reduced depth of crypts, which was more prominent in the distal part of the small intestine. Viral isolation attempts with primary cell cultures of chicken embryo kidney cells were negative. Examination by electron microscopy of negatively stained intestinal specimens revealed the presence of Reoviridae particles of 58.8 to 80 nm in diameter. Enzyme-linked immunosorbent assay results on the intestinal pools for mammalian and group A avian rotaviruses were negative. A statistically significant relationship was found for the presence of Reoviridae particles in the intestines of 10-21-day-old birds. Of the 7 most common pathological conditions analyzed, 2, rickets and intestinal changes (thin-walled ballooning intestine with watery contents), showed a statistically significant association with the presence of Reoviridae particles. PMID- 2562200 TI - Prospected randomized study of two Y devices in continuous ambulatory peritoneal dialysis (CAPD). AB - To evaluate acceptability, safety, and efficacy of a Y set with two short branches (TAS) filled with electrolytic chloroxidizer solution during the dwell time, 60 patients were randomly allocated to be treated with the traditional Y set (TCS) or with the TAS. Twenty-three were new patients whereas the remaining 37 were patients already on continuous ambulatory peritoneal dialysis (CAPD) with the TCS. The follow-up was 416.5 months in the control group and 387.4 months in the test group. During the study period there were 6 peritonitis episodes in each group with an incidence of 1 episode every 69.4 patient-months in the control group and 1 episode every 64.6 patient-months in the test group. Twenty-four patients (80%) in the control group and 27 (90%) in the test group were free from peritonitis. The probability to remain free from peritonitis was respectively 87% and 83% in the test group and in control group after 12 months, 70% and 78% after 21 months. Seventy-nine percent of the patients who used both systems preferred the TAS for better handling, lower encumbrance, and major safety. One patient preferred the TCS, three patients did not find any differences between the two devices. PMID- 2562201 TI - Changes in activity of selected lysosomal enzymes in peritoneal macrophages of renal failure patients on peritoneal dialysis. AB - Activity of acid phosphatase (AP), beta-glucuronidase (GR), N-acetyl-beta-D glucosaminidase (GZ), and peroxidase (P) was assessed using a semiquantitative cytochemical method in peritoneal macrophages of 30 patients with end-stage renal failure treated by intermittent peritoneal dialysis and of 30 control patients with normal renal function. The dialysed patients showed a significantly higher activity of GR and P at the beginning of the treatment as compared with the respective activities observed in the control group and a further significant rise of these activities after 4 months of dialysis. Activity of AP at the beginning of the treatment was insignificantly lower than in the control group and the difference became significant at the end of the investigated period. There was no significant difference between the dialysed patients and the control group in the activity of GZ assessed at the beginning of the dialytic treatment and after 4 months of dialysis. A significant decrease in that activity was, however, observed in the course of dialysis. PMID- 2562202 TI - [Uveitis of viral origin]. AB - Most well-defined viral uveitis cases are due to herpes viruses. Thus, HSV and VZV may cause a chronic anterior uveitis, usually with other characteristic signs. Cytomegalovirus causes a necrotic and haemorrhagic retinitis in immunosuppressed patients. Acute retinal necrosis also seems to be caused by members of the herpes virus family. Many cases of intraocular inflammation have been described in association with viral diseases, but this does not necessarily imply the presence of virus in the eye. PMID- 2562203 TI - [Cardiopulmonary resuscitation: acid-base alterations and alkalizing therapy]. AB - It is generally believed that metabolic acidosis prevails during cardiac arrest. However, recent experimental and clinical studies have demonstrated that respiratory acidosis in mixed venous blood and respiratory alkalosis in arterial blood with only minor increases in lactic acid characterize the early acid-base changes that follow cardiac arrest and cardiopulmonary resuscitation (CPR). While continued CO2 production with critical reduction in systemic perfusion explains the accumulation of CO2 in the venous side, the reduction of pulmonary blood flow with maintenance of constant minute ventilation explains the decreases in expired CO2 and therefore arterial PCO2. In the heart, marked increases in CO2 tension and lactic acid are associated with dramatic decreases in myocardial pH with consequent depression of contractile function. Administration of sodium bicarbonate, however, neither increases resuscitability nor improves long term outcome. Moreover, adverse effects stemming from increases in plasma osmolality, increases in hemoglobin-O2 affinity, induction of alkalemia and generation of CO2 are potentially deleterious for myocardial and cerebral function. Consequently, the American Heart Association has recently discouraged the routine administration of bicarbonate during the initial 10 minutes of CPR in which interventions with proven efficacy such as artificial ventilation, precordial compression, electric defibrillation and epinephrine administration take place. Alternative experimental buffer therapy with agents that consume CO2 have also failed to alter the outcome of cardiac arrest. PMID- 2562204 TI - The radiology of phyllodes tumour. PMID- 2562205 TI - The anticoagulant activity of heparin: measurement and relationship to chemical structure. AB - For many years the anticoagulant activity of heparin has been estimated by coagulation assays, in which the prolongation of clotting times by heparin is measured under various conditions. More recently, assays have been developed which measure the inhibitory action of heparin on isolated coagulation enzymes, notably Factor Xa and thrombin, using specific amidolytic peptide substrates. The anticoagulant activity of heparin arises primarily from its ability to bind to antithrombin III (AT III), altering the conformation and enhancing the activity of this major protease inhibitor. Passage of heparin through an immobilised AT III column yields two fractions: a high affinity fraction with 300-350 iu mg-1 anticoagulant activity, comprising one-third of the total, and a low affinity fraction with an activity of less than 10 iu mg-1, comprising the remaining two thirds. Studies in several laboratories have demonstrated that a specific pentasaccharide sequence is required for AT III binding. The authors have shown that the presence or absence of this sequence can be detected by high-field proton NMR, thus providing a semi-quantitative method for a functionally important group. A second major influence on anticoagulant activity is molecular weight distribution. Studies in the authors' laboratory on a series of fractions of 5000-35,000 showed that whereas anticoagulant activity in APTT clotting assays decreased with decreasing molecular weight (Mr), activity in anti-Xa assays was maintained or increased in the low Mr fractions. However, in vivo studies showed that high affinity fragments with anti-Xa activity only were poor antithrombotic agents. It appears that the presence of the AT III binding site alone is not sufficient for full antithrombotic activity; an extra length of polysaccharide chain of at least 15 residues is required. Molecular weight distribution is readily assessed by HPLC, although the lack of suitable reference materials hampers assignment of absolute molecular weights. Important determinants of anticoagulant activity can now be assessed by physicochemical techniques but, at present, these techniques are not precise enough to replace anticoagulant assays as predictors of in vivo behaviour. PMID- 2562206 TI - Interferon-alpha: a gene family in therapeutic use. AB - Several variants of interferon-alpha (IFN-alpha) were isolated and purified to homogeneity. They differed to various degrees in biological properties. However, three IFN-alpha 2 variants showed only minor differences from a variant called IFN-alpha 88 with regard to their ability to inhibit growth and to bind to specific receptors, tested on Daudi cells. Two monoclonal antibodies were studied, which showed overlapping specificity for at least one peptide obtained after HPLC separation of tryptic digests. The monoclonal antibodies could discriminate between sequence differences to a much higher degree than the receptor on Daudi cells. It is concluded that the receptor is degenerate and binds well to different structural variants of IFN and that for therapeutic use, several of the variants will probably have the same biological potency. PMID- 2562207 TI - Detection of bovine viral diarrhea virus by spot hybridization with probes prepared from cloned cDNA sequences. AB - A cytopathic strain of bovine viral diarrhea virus (BVDV) was purified from infected cell culture fluids by isopycnic density-gradient centrifugation. Genomic RNA was extracted and tailed with adenine residues at the 3' end with poly-A polymerase. Double-stranded complementary DNA (cDNA) was synthesized, using the poly-A-tailed RNA as a template and oligo-dT as a primer, and then cloned into the pUC9 plasmid. Virus-specific cDNA sequences, varying in length from 0.5 to 2.5 kilobases (kb), were obtained. One BVDV-specific sequence of cloned cDNA, 1.1 kb in length and with an internal Pst I restriction endonuclease cleavage site, was selected for use as a probe. The cloned cDNA insert was removed from the plasmid either with or without flanking plasmid sequences and labeled with 32P-nucleotides by nick translation for use as hybridization probes for BVDV. The performance of probes of smaller fragments of the insert was compared to that of the intact sequence in hybridization assays. In addition, 2 methods of specimen preparation were compared to establish optimum parameters for hybridization. The hybridization assay was 10-100 times more sensitive than infectivity assays for BVDV in infected cell cultures. Freezing of specimens reduced by 10-fold the sensitivity of hybridization for BVDV target sequences. The probes prepared from the cloned cDNA hybridized with all cytopathic and noncytopathic BVDV strains tested but not with uninfected cell cultures, cellular ribosomal RNA, bovine coronavirus, bluetongue virus, or bovine adenovirus 3. Probes prepared with native plasmid DNA did not hybridize with BVDV or uninfected cell cultures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562208 TI - Rapid fluorescence detection of in situ hybridization with biotinylated bovine herpesvirus-1 DNA probes. AB - The use of biotinylated DNA hybridization probes for clinical detection of bovine herpesvirus-1 was investigated. Biotinylated DNA hybridization probes were prepared from bovine herpesvirus-1 DNA purified from infected cell cultures. The viral DNA was nick translated in the presence of biotin-dUTP with DNA polymerase to incorporate biotin into the newly synthesized strand. The probe was tested for specificity in in situ hybridization assays with bovine herpesvirus-1 DNA. Hybridization was detected using avidin-fluorescein single sandwich systems and an avidin-globulin with anti-globulin-fluorescein double sandwich system. Hybridization was detected by specific fluorescence of infected cells. Fluorescence was present only in bovine herpesvirus-1-infected cell culture and not in noninfected cell culture or cell cultures infected with several other viruses. The assay was performed in 6 hr. PMID- 2562209 TI - Use of an immunoperoxidase test for the detection of bovine herpesvirus-1 in aborted fetal tissue. AB - An indirect immunoperoxidase (IP) procedure using a specific monoclonal antibody and an avidin-biotin-peroxidase complex was developed and applied to detect virus antigen in formalin-fixed, paraffin-embedded tissue sections. This IP procedure was compared with currently used diagnostic tests for detection of virus-induced abortions caused by bovine herpesvirus-1 (BHV-1). The IP procedure was applied to detect BHV-1 antigen in sections of liver and lung from 87 aborted fetuses. Sixteen of these cases were positive for viral antigen by IP staining. Sections from both liver and lung were positive in 15 of the 16 cases. A fluorescent antibody test (FA), which was applied to acetone-fixed frozen sections of liver and lung, gave positive results on 12 of the 87 fetuses, 11 of which were also positive by IP. Seven of the 12 FA-positive cases were positive on both sections of liver and lung. When FA and IP were compared. FA had a sensitivity of 67% and IP had a sensitivity of 94%. Virus was isolated from one of the 67 cases tested. The tissues in which antigen was most frequently detected by IP were liver, lung, and kidney. Distinct multifocal staining was seen in positive sections of all these tissues. PMID- 2562210 TI - Detection of bluetongue virus in bovine fetuses using the avidin-biotin complex immunoperoxidase method. AB - The avidin-biotin complex immunoperoxidase technique was adapted for use in detecting bluetongue virus (BTV) antigens in BTV serotype 11-infected bovine fetuses. Fetuses were infected with BTV serotype 11 at 120 days of gestation and then removed 20 days later by Cesarean section. Blood and tissue samples were collected from each animal and used for virus isolation in embryonated chicken eggs, the immunofluorescent antibody test, and the avidin-biotin complex test. The avidin-biotin complex method successfully identified BTV antigens in both fresh and autolyzed fetal brains. Thus, the avidin-biotin complex immunoperoxidase method has potential as a possible procedure for diagnosing bluetongue disease in aborted bovine fetuses. PMID- 2562211 TI - Comparison of diagnostic tests for the detection of equine infectious anemia antibody. AB - Two diagnostic tests are approved for detecting antibody to equine infectious anemia virus: the agar-gel immunodiffusion (AGID) test and the competitive enzyme linked immunosorbent assay (ELISA). A total of 420 sera from National Veterinary Services Laboratories check sets were tested with the AGID and competitive ELISA. A 100% correlation was obtained. The AGID and competitive ELISA were further used to test difficult samples with low levels of equine infectious anemia antibody (weak positives). A third test (Western blot) was also used with these weak positive samples to resolve any discordant results. PMID- 2562212 TI - Diagnostic investigation of bovine viral diarrhea infection in a Minnesota dairy herd. AB - Bovine viral diarrhea (BVD) virus infection was diagnosed in neonatal calves with enteritis. Successful diagnostic procedures included direct immunofluorescence of frozen tissue sections, histopathology, and virus isolation. Virus isolation from buffy coats and serum was successful in detecting infected animals, whereas direct immunofluorescence of buffy coat samples was found to be less reliable. Virus was not isolated from any fecal samples. Booster vaccinations and the culling of animals shedding virus resulted in improved calf viability in this herd. It is suggested that procedures for the diagnosis of BVD virus infection should always be included in the diagnosis of neonatal calf enteritis. PMID- 2562213 TI - Detection of rotavirus in fecal samples from calves by a cell culture indirect immunofluorescence, an Ag-capture ELISA, a tissue culture ELISA, and a commercial Ag-capture ELISA. PMID- 2562214 TI - A novel bovine rotavirus electropherotype from outbreaks of neonatal diarrhea in Utah beef herds. PMID- 2562215 TI - The effect of diet on tumor development in animals. PMID- 2562216 TI - Prophylaxis of AIDS-related opportunistic infections (OIs). PMID- 2562217 TI - Prognostic significance (relapse, non-relapse) of nuclear shape parameters in lymph node negative breast cancer. AB - A series of 15 postmenopausal patients with stage I invasive breast cancer (pT1 2, pNO, cMO) of uniform histologic type (ductal, NOS) was selected from the Basel Cancer Registry for retrospective analysis. A group of seven patients with a minimal disease-free survival of 9 years was compared to a group of eight patients with clinical and/or histopathological relapse within 8 years of mastectomy. Computer-assisted morphometry of cancer nuclei in routine histological material was used to investigate possible correlations of clinical outcome with nuclear size and shape parameters. Significant differences were found for certain shape descriptors by nonparametric statistical analysis. Stepwise discriminant analysis was applied to find the best combination of parameters separating the two prognostic groups. Application of the 'leaving one out method' showed correct classification in 6 of 8 recurrent cases and in all non-recurrent cases. The number of patients is small, therefore the results should be carefully interpreted. PMID- 2562218 TI - HIV trans-activation and transcription control mechanisms. AB - Promoter-specific transcription factors, whose function was once thought to be limited to initiation, are now known to have more diverse roles in RNA metabolism, including the cellular localization of transcripts and the integration of RNA initiation with attenuation and RNA 3' end formation. The human immunodeficiency viruses (HIV-1 and HIV-2) provide a useful system to study such proteins, since distinct DNA and RNA elements downstream of the site of transcription initiation act in conjunction with the promoter to regulate the induction and attenuation of RNA synthesis. Sequences corresponding to the 5' untranslated leader of HIV-1 and HIV-2 harbor at least three distinct elements: (i) a DNA domain that binds LBP-1, a cellular activator of initiation; (ii) a structured RNA element critical for the function of the HIV-1 trans-activating protein, Tat; and (iii) an RNA element required for the production of attenuated RNAs from the basal (uninduced) promoter. These attenuated leader RNAs seem to be created in vitro by stalled RNA polymerase II complexes that may be uniquely capable of rapidly processing RNA. Tat-mediated increases in steady-state levels of viral transcripts appear from nuclear run-on experiments to involve a control mechanism at both initiation and early post-initiation steps. Studies that implicate a role for Tat in post-transcriptional control suggest the existence of a mechanism for the coordination of eukaryotic transcription and translation, possibly through the assembly of nuclear regulatory factors at the 5' end of the RNA. PMID- 2562219 TI - E. coli ribosomes re-phase on retroviral frameshift signals at rates ranging from 2 to 50 percent. AB - Many retroviruses express gag-pol or gag-pro-pol polypeptides by coupling their translation from overlapping reading frames with -1 ribosomal frameshifts. Here, we show that the well-known ribosomal frameshift signals found in retroviral mRNA will provoke Escherichia coli ribosomes to shift frame in the same manner as their eukaryotic counterparts. Ribosomes of E. coli respond in vivo to both the tandem slippery codons present at the retroviral frameshift site and the 3' flanking sequence. Slight alteration of the mouse mammary tumor virus gag-pro frameshift site from A-AAA-AAC to A-AAA-AAG boosts the level of frameshifting in E. coli to over 50%. This suggests that A-AAA-AAG, and its slippery relatives, may be utilized by E. coli genes as sites of high-level ribosomal frameshifting. This observed conservation of response to retroviral frameshift signals affords new avenues to dissect the mechanism of ribosomal frameshifting evoked by these mRNA sequences. PMID- 2562220 TI - Jun DNA-binding is modulated by mutations between the leucines or by direct interaction of fos with the TGACTCA sequence. AB - Mutations between the leucines of the "leucine zipper" domain of Jun D can either decrease (Asn 301 to Ala) or increase (Thr 307, Ala 308, to Glu, Val) homodimer formation and specific binding to DNA even though such changes do not modify the predicted alpha-helical structure of this region. As shown previously, addition of Fos strongly increases the affinity of Jun for DNA by forming a heterodimer. The jun down mutation (Asn 301 to Ala) also diminishes DNA binding by the Fos-Jun D heterodimer. These data strongly support the coiled coil conformation of this region where residues adjacent to the leucines are also important for dimer formation. Ultraviolet cross-linking experiments have shown that both Fos and Jun directly contact the TGACTCA palindromic sequence defined as a TPA (12-O tetradecanoyl phorbol-13-acetate) response element or TRE. Both Jun homodimers and Jun-Fos heterodimers bind this TRE as well as the cAMP responsive element (CRE or TGACGTCA) with comparable affinities. While strong c-Jun or Jun D binding requires a perfect palindrome, Jun-Fos complexes can also efficiently recognize sequences where the right half of the palindrome is less conserved (TGACTAA or TGACGCA). PMID- 2562221 TI - Differentiation and oncogenesis: phenotypically distinct lens tumors in transgenic mice. AB - We report on two lines of transgenic mice that express a murine alpha A crystallin/SV40 tumor antigen fusion gene in the eye lens. The alpha T1 line develops fast growing, poorly differentiated lens tumors, whereas the alpha T2 line produces lens tumors that are slow growing and well differentiated. There is a striking difference between these two lines in the temporal and spatial patterns of tumor antigen expression during initial lens development. In the alpha T1 line, the transgene is expressed very early in development in most lens cells, and no primary fiber differentiation takes place. In the alpha T2 line, transgene expression occurs after primary fiber formation has been initiated, and is restricted to differentiating fiber cells. The anterior epithelium from both alpha T lines undergoes normal development and remains morphologically normal until after birth, although in alpha T1 mice, these anterior cells produce considerable amounts of SV40 tumor antigens. This suggests that the state of differentiation of the lens cell plays an important role in its response to oncogene products. PMID- 2562222 TI - Homologous recombination involving small single-stranded oligonucleotides in human cells. AB - Gene modification by homologous recombination is one of the techniques that may eventually be used in gene replacement therapy. We tested whether small, synthetic single-stranded oligodeoxynucleotides are capable of participating in homologous recombination in human cells. A plasmid carrying a mutant neomycin phosphotransferase (neo) gene was cotransfected with a 40-nucleotide single stranded oligomer that contained the wild-type neo gene sequence into human cells. Cells expressing neo were selected in the antibiotic G418. These cells contained wild-type molecules, which resulted from recombination between the two molecules. The results indicate that this approach may be useful in correcting or introducing single point mutations into the genomes of mammalian cells. PMID- 2562223 TI - [Seroepidemiologic study of hepatitis A virus among children of Maracaibo, Venezuela]. AB - With the purpose of determining the incidence of Hepatitis A virus infection, we determined the presence of antibodies against this virus (anti-VHA) with the ELISA technique in 209 children from Maracaibo city of different socioeconomic levels, ages ranging from 6 months to 12 years. At the same time, we investigated the incidence of a recent infection through IgM antibodies for Hepatitis A (anti VHA IgM) using the same ELISA technique. We found a global positive incidence of 46.4% for all ages. Individual ages studied showed 31% seropositivity in children less than one year old and a higher seropositivity in children between 9 and 11 years old (50-76.4%). There were noticeable differences in seropositivity in relation to social economic condition observing a higher percentage in children of low social economic level. A recent infection was demonstrated after the second year of life (60%) and a variable percentage through all pediatric ages studied. There was no recent infection detected in children under one year of age which suggests that antibodies anti-VHA obtained transplacentally can last at least for the first year of life as it does with other virus. PMID- 2562224 TI - Use of in situ hybridization for the detection of foot-and-mouth disease virus in cell culture. AB - Biotinylated complementary DNA (cDNA) and RNA probes were prepared from a specific and highly conserved section of the foot-and-mouth disease virus (FMDV) genome coding for the RNA-dependent RNA polymerase. Hybridization was conducted on FMDV-infected, bovine enterovirus (BEV)-infected, and noninfected swine kidney cell cultures. The detection system utilized the enzyme system streptavidin alkaline phosphatase, the substrate phosphate, and the chromogen nitroblue tetrazolium. Intense cytoplasmic granular staining was present at 2 and 4 hr postinfection (hpi), with less staining observed at 24 hpi. The staining was specific for FMDV, as indicated by a lack of staining of noninfected cells and BEV-infected cells. With the RNA probe, positive cells were detected up to the highest viral dilution assayed, which was approximately 96 TCID50. The cDNA probe was slightly less sensitive, detecting positive cells at 10-fold lower dilutions. This technique could prove useful in the diagnosis of foot-and-mouth disease in animals or in the detection of FMDV in biologics submitted for importation. PMID- 2562225 TI - Feline immunodeficiency virus infection in a population of pet cats from southeastern Florida. AB - Blood samples from 95 randomly selected pet cats that were brought to veterinarians in southeastern Florida were tested for antibodies to feline immunodeficiency virus (FIV). Virus-specific antibodies (indicative of virus infection) were found in 8 of the 95 (8.4%) cats tested. All of the virus infected cats were males (statistically significant, P less than or equal to 0.016) and were at least 1 year of age. The 3 most severely ill cats infected with FIV were also infected with feline leukemia virus. PMID- 2562226 TI - Diarrhea associated with Clostridium perfringens type A enterotoxin in neonatal pigs. PMID- 2562227 TI - A molecular basis for intersystem communication between the immune and neuroendocrine systems. PMID- 2562228 TI - [Proton potential on the inner mitochondrial membrane and its role in mitochondrial processes]. PMID- 2562229 TI - [Transport of calcium ions through the cell membrane]. PMID- 2562230 TI - [The role of active forms of oxygen in cell differentiation, neoplasm promotion and aging]. PMID- 2562231 TI - Neurovascular activation requires conduction through vessels. AB - In experiments on rats, a hind leg was transected except for the femoral nerve, artery and vein. The femoral nerve was stimulated by electrical pulses, with one electrode attached to the nerve in the amputation gap and another placed in the inferior caval vein. At each pulse of stimulation, contraction could be recorded in femoral muscles. Ligation of the femoral vessels suspended the contractions within a fraction of a second; contractions resumed when ligation ended. Propagation of neural stimuli to femoral muscles therefore required an intact electrical communication through associated vessels. This is possible because conducting blood plasma and its capillary junction with the interstitial fluid form an "external" closed circuit branch of low resistance with the neuromuscular unit. In testing the interstitial tissue fluid as an alternative to the vascular "outer" communication, a higher voltage of stimulation was required for muscle contractions. PMID- 2562232 TI - Magnetic resonance of water protons in fresh human blood plasma. AB - Spin-lattice (T1-) relaxation times of fresh human blood plasma at 13.2 MHz and 29 degrees C ranged from 1263 milliseconds (msec) to 1709 msec. Spin-spin (T2-) relaxation times of those samples were between 446 msec and 753 msec. Proton magnetic resonance (p.m.r.) phantoms of such blood plasma were made with ferric chloride and corn starch in dilute hydrochloric acid, and also in dilute sulfuric acid. Their Fe3+ ion concentrations approximated 138 micrograms (micrograms) per deciliter (dl). Both T1 and T2 of any of these p.m.r. phantoms were within limits of those described above for fresh human blood plasma. Lowering of the concentration of the Fe3+ ion--in an experimental corn starch solution--was manifested in longer T1. PMID- 2562233 TI - The effect of magnesium valproate on plasma ACTH concentrations in Nelson's syndrome. AB - Sodium valproate, a gamma-aminobutyric acid (GABA) agonist, was found to decrease plasma ACTH concentration in some cases of Cushing's disease and Nelson's syndrome. In this study we have investigated the influence of magnesium valproate (MV), a newly introduced salt of valproic acid, on plasma ACTH levels in 8 patients with Nelson's syndrome. The daily dose, 1200 mg of MV, significantly decreased plasma ACTH level at 10 p.m. compared with placebo. A single dose of 400 mg of MV, led to a reduction in plasma ACTH concentration only in two out of seven patients during a four-hour observation. The fall in plasma ACTH level in the same patients at 10 p.m., after the next two doses of this drug, suggests that single dose may be insufficient for introducing GABA-dependent reduction in ACTH release. During a long-term therapy with MV, in all three patients investigated a marked decrease in plasma ACTH was observed. Our results suggest that magnesium valproate may be useful during chronic therapy in some patients with ACTH hypersecretion. PMID- 2562234 TI - [Elaboration of an immunosorbent for the purification of porins from Salmonella typhi 9, 12, Vi:d]. AB - The current work was undertaken to purify porins of Salmonella typhi, which are outer membrane proteins (OMPs) that induce protection in mice against challenge with the bacteria in mucin. OMPs, isolated with a non-ionic detergent, had a 4% contamination with LPS (endotoxin) and molecular sizes ranging from 17 to 70 KDa. Porins (Mw 38-41 KDa) were isolated from OMPs preparative SDS-PAGE. Anti-porins antisera were raised in rabbits and specific IgG was purified, which was coupled to Sepharose-CNBr. This immunosorbent was used to purify LPS-free porins. PMID- 2562235 TI - Occurrence of human papillomavirus type 16 DNA sequences and c-myc oncogene alterations in uterine-cervix carcinoma. AB - Using genetic engineering and molecular biology techniques, we have examined sixteen human carcinomas in the uterine-cervix tumors (the most frequent tumor in Mexico, representing 34% of malignant tumors in women), for the presence of Human Papillomavirus type 16 (HPV-16) DNA sequences and possible alterations of the cellular myc (c-myc) proto-oncogene. In this study we have analyzed cervical carcinomas from patient with clinical stage II. We detected in 31% of these samples, the presence of HPV-16 sequences (2-100 copies). In addition, an elevated amplification (up to 80-fold in one tumor) and/or rearrangement of the c myc oncogene was detected in most tumors (more than 90% of the samples). These results suggest that either c-myc oncogene and/or HPV-16 could play an important role in the development of uterine-cervix carcinoma. PMID- 2562236 TI - Morpheus: a conformation-activity relationships and receptor modeling package. AB - Our molecular modeling software package, MORPHEUS, allows the study of the interactions between biologically active molecules and their receptors. The package is capable of exploring the multidimensional conformational space accessible to each molecule of the data set under study. By specifying distance constraints or hypothetical receptor binding points, the package is able to filter the biologically accessible conformations of each active compound and deduce a three-dimensional model of the binding sites consistent with the properties of the agonists (or antagonists) under scrutiny. The electrostatic potentials in the environment of a putative binding site can also be investigated using the MORPHEUS package. The molecular modeling module CRYS-X, which is written in FORTRAN 77 for IBM PC machines, is capable of building, displaying and manipulating molecules. PMID- 2562237 TI - Diagnostic value of salivary gland scintigraphy with technetium-99m pertechnetate. PMID- 2562238 TI - [Electron microscopic study on malignant fibrous histiocytoma induced by local application of DMBA heterotransplanted in nude mice]. AB - Though Malignant Fibrous Histiocytoma (MFH) has been regarded as a relatively common neoplasm of the mesenchymal soft tissues, the issue of character is largely unresolvable at present. Our department formerly reported that produced experimental MFH by administering 9, 10-dimethyl-1, 2-benzanthracene (DMBA) in male hamsters. In this study I have examined the fine structure of transplantation and heterotransplantation of MFH. The following results were obtained. 1. Five different types of cells were found in the MFH: fibroblast-like cells, histiocyte-like cells, undifferentiated cells, multinucleated giant cells and xanthomatous cells. 2. In histiocyte-like cells, nonphagocytic and actively phagocytic histiocyte-like cells could be distinguished. 3. The multinucleated giant cells could be found two types, fibroblast-like and histiocyte-like. 4. In transplantation, the histology of the original tumor was retained. 5. In heterotransplantation, the tumor was composed mainly of nonphagocytic histiocyte like cells and undifferentiated cells. 6. The possibility is suggested that both principal cell types in this tumor may derive from the same undifferentiated stem cells. PMID- 2562239 TI - [Establishment and characterization of neoplastic cells derived from a hamster malignant fibrous histiocytoma induced by DMBA]. AB - The malignant fibrous histiocytoma (MFH) induced by 9,10-dimethyl 1,2-benz anthracene (DMBA) in the hamster, was prepared for tissue culture work. By applying the soft agar method, a clone was isolated. The cloned cell line was proved to be homogenous in morphology and ultrastructure, showing fine structure similar to histiocyte. The cells were positive for acid phosphatase, alpha naphthyl acetate esterase and lysozyme in cytochemistry. Phagocytic activity of the cells tested by using carbon particles was found in the greater part of them. Moreover, inoculation of the cells into nude mice resulted in production of tumors belonging to MFH. Even after a long-term cultivation, no notable morphologic alternation was found and the cells retained histiocytic functions. PMID- 2562240 TI - [Clinical and therapeutic profile of 3 cases of cryptococcal meningitis in patients with AIDS]. AB - We report three cases of cryptococcal meningitis in patients with AIDS observed in our Institution. In addition, we discuss antifungal treatment during and after the acute phase of the disease and the use of fluconazole as a prophylactic treatment of disease relapse. PMID- 2562241 TI - [Cytomegalovirus infections in pregnant women]. PMID- 2562242 TI - [Physicochemical characteristics of new reinforcement ceramic implant]. AB - Investigation of the physicochemical characteristics of reinforcement ceramic implant materials which have needle crystal produced the following results. 1. Simple diopside and hydroxyapatite with diopside have enough dynamic intensity of both the implant in point of bending intensity and the breaking toughness. 2. Simple diopside and hydroxyapatite with diopside go well as biomechanics because of their Young's modulus being close to the surrounding bone. 3. Diopside in pseudo-body fluid developed hydroxyapatite on surface like bioglass, and this resulted in high possibility of direct bond with bone. 4. The results of this study indicate that the diopside and hydroxyapatite with diopside have enough dynamic intensity and are bioactive dental implant materials. PMID- 2562243 TI - Immunoglobulin VH gene expression in human B cell lines and tumors: biased VH gene expression in chronic lymphocytic leukemia. AB - We have studied frequencies of VH gene utilization in a panel of monoclonal Epstein-Barr virus (EBV)-transformed B cell lines derived from human adult and fetal tissues as well as in monoclonal B cells obtained from fresh chronic lymphocytic leukemia (CLL) samples. The results show that IgM-secreting EBV cell lines from both fetal and adult tissues utilize VH genes from particular families roughly in proportion to estimated family size, suggesting that the repertoire of sigM-positive B cells in both fetal and adult organs is 'normalized' with respect to the V(H) gene family. In contrast, we find a highly biased pattern of VH gene expression in CLLs. The significance of these findings is discussed in the context of mechanisms that could be involved in normal B cell repertoire development and in the process of malignant transformation of precursors of CLL. PMID- 2562244 TI - Evidence for differential expression of CD45 isoforms by precursors for memory dependent and independent cytotoxic responses: human CD8 memory CTLp selectively express CD45RO (UCHL1). AB - On the basis of differential CD45 expression, human T cells can be separated into approximately reciprocal populations. The mAb UCHL1 detects a 180 kd molecular mass isoform, CD45R0. The CD45RA cluster of mAbs reacts with CD45 isoforms of 205 and 220 kd molecular mass. These reagents subdivide both CD4 and CD8 T cell populations. CD4 T cells that proliferate in response to memory-dependent (recall) antigens have been shown to selectively express CD45R0. We extend these observations to a model for cytotoxic responses that allows the functional analysis of CD8 T cells with differential CD45 expression. Precursors for allo specific CTL responses are readily detectable in CD45R0 as well as CD45RA populations. In contrast, we find memory CTLp greatly enriched among CD45R0 cells. In combination with earlier work, these results suggest that differential expression of CD45 isoforms is associated with memory formation for different classes of immune responses in both major T cell lineages. PMID- 2562245 TI - Predominant use of a particular alpha-chain in suppressor T cell hybridomas specific for keyhole limpet hemocyanin. AB - We isolated and sequenced the rearranged genomic variable (V) and joining (J) gene segments of T cell receptor alpha-chain gene from two independent keyhole limpet hemocyanin (KLH)-specific suppressor T cell (Ts) hybridomas (BW5147 x C57BL/6 KLH-Ts). These nucleotide sequences were compared with those of germline DNA from kidney and also with cDNA of alpha-chain (VJ alpha 281) previously isolated from Ts hybridoma (34S-281) with KLH/H-2b Ts activity. The entire V alpha and J alpha sequences of all three Ts hybridomas were exactly identical and were encoded by the germline V alpha and J alpha gene segments without any mutations, except for 2-nucleotide deletions from both the 3' end of V alpha and 5' end of J alpha gene segments, respectively, and a 1-nucleotide (guanine) insertion in the junctional (N) region which was not encoded by the germline gene. Six additional KLH-Ts hybridomas, further analyzed, also possessed the same alpha-chain, indicating the preferential usage of the particular alpha-chain in these hybridomas. As chromosome analysis demonstrated a different pattern in each clone, these hybridomas appear to be independent. More surprisingly, 0.5-1.5% of the total functional T cell alpha-chain mRNA in the thymus and spleen of unprimed C57BL/6 mice was found to be of this particular alpha-chain. These results suggest that the repertoire of KLH-Ts is strictly limited. PMID- 2562246 TI - Conformation and stability of Sendai virus fusion protein. AB - The conformation and stability of Sendai virus fusion (F) protein were studied by circular dichroism spectroscopy, and the protein predictive models of Chou and Fasman and Robson and Suzuki were used to elucidate the secondary structure of Sendai virus F protein. The F protein conformation is predicted to contain 33% alpha-helix, 53% beta-sheet and 15% beta-turn by the Chou and Fasman model, and 30% alpha-helix, 55% beta-sheet, 9% beta-turn and 7% random coil by the Robson and Suzuki model. C.d. studies of F protein purified in the presence of the non ionic detergent, n-octylglucoside, indicated the presence of 49% alpha-helix and 31% beta-sheet at pH 7.0, 54% alpha-helix and 28% beta-sheet at pH 9.0 and 50% alpha-helix and 23% beta-sheet at pH 5.4. A small change in conformation of the protein occurred when the pH was titrated from 7.0 to 5.4 and from 7.0 to 9.0 and a more pronounced conformational change occurred when the pH was changed from 9.0 to 5.4. The F protein in 0.2% n-octylglucoside was resistant to denaturation by 4 M guanidine hydrochloride, the reducing agent 20 mM mercaptoethanol, and to increases in temperature from 5 to 80 degrees C. Monoclonal anti-F protein antibody showed an increased binding to whole virus when the pH was changed from 7.0 to 9.0. The antibody binding was decreased when the pH was shifted from 9.0 to 5.4 Maximum haemolytic activity was observed with virus that was preincubated at pH 8.0.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562247 TI - Chitin gels. AB - Chitin dissolved in N,N-dimethylacetamide, N-methyl-2-pyrrolidone and their mixed solvents in the presence of 5% LiCl was treated with acetic anhydride-pyridine, and the mixture solution was heated at 100 degrees C for 6 h to give a partially O-acetylated chitin gel. Chitin dissolved in these solvents in the presence of 5% LiCl was mixed with pyridine, and the mixture solution was heated at 60 degrees C for 5 h to give a chitin gel. Both the gels were rigid and transparent, and their properties and the rate of the hydrolysis of the chitin xerogel by hen-egg white lysozyme were essentially similar to those of N-acetylchitosan gel prepared by chemical N-acetylation of chitosan. PMID- 2562248 TI - Pulmonary complications of AIDS. PMID- 2562249 TI - Other virus infections in AIDS. I. Cytomegalovirus. PMID- 2562250 TI - Other virus infections in AIDS. III. Epstein-Barr virus. PMID- 2562251 TI - Deficiency of the core proteins of dermatan sulphate proteoglycans in a variant form of Ehlers-Danlos syndrome. AB - A variant form of Ehlers-Danlos syndrome with single adrenocorticotrophic hormone (ACTH) deficiency and mild diabetes mellitus was studied by immunohistochemical and biochemical analyses of the connective tissue. The patient exhibited features characteristic of the Ehlers-Danlos syndrome, such as fragility, hyperextensibility, and bruisability of the skin and large veins were readily seen, but no hypermobility of any joint was detectable. Biochemical studies of the connective tissue confirmed that the apparent synthesis of collagens was within normal limits, but the deposition of the major proteoglycan in the skin of this patient was markedly reduced, mainly because the synthesis of core protein of dermatan sulphate proteoglycan was defective. PMID- 2562252 TI - Chronic infectious mononucleosis; a case report. AB - A 39-year-old woman had an adenitis colli followed by anaemia, splenomegaly, atypical lymphoid cells in blood with increased B-lymphocytes, reduced T suppressor/cytotoxic cells, increased polyclonal IgM, high titres of EB VCA IgG, EB EA IgD&R +/- and EBNA IgG-. The disease progressed slowly for 2 years, splenectomy was followed by clinical improvement; spleen morphology was compatible with a benign disease of viral origin. PMID- 2562253 TI - Phthaloyl-glycylP-isoleucyl-tryptophan benzylamide is a potent inhibitor of human skin fibroblast collagenase with a Ki of 25 nM. PMID- 2562254 TI - [Influence of acid buffer on the crystallinity of enamel apatite]. AB - The affects of acetic buffers on the crystallinity of an enamel apatite were examined in the present study. Dual aspects, crystallite size and lattice imperfection related to the crystallinity were analyzed by the process of Variance and Fourier analysis based on the X-ray diffraction line profiles. For this purpose both enamel apatite and synthetic hydroxyapatite were analyzed through acetic acid buffer decalcification, and comparison was made. In addition to the analysis, morphological change of the apatite after acid decalcification was observed by a transmission electron microscope (TEM). The results obtained in the present study were as follows: 1) Wedge-shaped defects formed on a prism plane were observed. In the further stage, wedge-shaped defects were also formed on a basal plane, and it proceeded into the center of the crystallite and consequentry slit similar to defected image was observed. 2) Lattice imperfection of the enamel and synthetic apatite were noticed both on the c-axis and a-axis. Among the factors which contributed to the variance, the "mistake" of lattice was much more likely than the crystallite size, and more mistakes in the direction of the a-axis rather than the c-axis was noticed. 3) A diminished crystallites size was found after acid treatment and diminishing on the c-axis was more apparent than the a-axis. 4) The crystallinity of the apatite was improved through acid treatment, and this improvement was attributed to a diminishing of mistakes. 5) Diminishing of mistakes through acid treatment was detected apparently along the c-axis. According to the results obtained in the present study, crystallite of enamel apatite showed decrease in size, although crystallinity through acid treatment was improved due to the diminishing of mistakes along the c-axis. This may be explained as following in that when apatite crystals were acid treated, the loci of lattice dislocation was selectively dissolved and consequently the more stable crystallite was left. PMID- 2562255 TI - [Comparative study of immunologic techniques applied to the detection of herpes simplex virus (HSV)]. AB - Indirect immunofluorescence (IF), immunoperoxidase (IP) and seroneutralization tests were compared for detection and identification or Herpes simplex virus (HSV), directly in tissue smears or in cells inoculated with clinical materials. Detection and clear differentiation between the two HSV serotypes were obtained using rabbit immunoglobulin cross-absorbed with heterologous virus antigen. Immunofluorescent staining with specific monoclonal antibodies is as sensitive and more rapid than standard Vero cell cultures, for the laboratory diagnosis of HSV. PMID- 2562256 TI - [Optimal detection of herpes simplex virus in clinical specimens preserved in transport media]. AB - The sensitivity of the methods used for HSV detection depends upon the stage and the transport of the collected lesions. High detection rates are achieved with early vesicular lesions rather with ulcers and scabs . Since HSV is heat-labile and to avoid loss of infectivity, specimens should be collected in transport medium and inoculated as soon as possible on the cell cultures. PMID- 2562257 TI - [Evaluation of molecular hybridization tests and DNA restriction enzyme analysis for the detection and typing of herpes simplex virus (HSV)]. AB - Restriction enzyme analysis of DNA and blot hybridization tests were applied for the identification of 90 herpes simplex viruses isolated. The results obtained by those methods correlated completely with typing by monoclonal antibodies in an indirect immunofluorescent assay. PMID- 2562258 TI - [Herpes simplex virus infections in Algiers]. AB - The purpose of this study was to determine the prevalence of herpes simplex virus antibodies among the population in Algiers. Anti-bodies to HSV1 are acquired rapidly between the ages of 1 and 6 years and 81.25% of the population is HSV1 seropositive by 15 years of age. Patients suffering from genital disorders possess HSV type 2 antibodies at a rate significantly higher (p less than 0.001) than in the control group. PMID- 2562259 TI - [Detection of rotavirus in the stools of infants aged 0-3 yr (study performed from July 1987 to May 1989)]. AB - 301 stools were collected during a period of 2 years from children aged 0-3 years, most of them presenting an acute gastro-enteritis; search for rotavirus was carried out using ELISA and latex techniques, together with coproculture. 44 stools were positive (14.61%), 18.68% of these positive cases were in the age groups 0-6 months. The rotavirus gastro-enteritis affects boys (63.63%) two times more often than girls. It is frequent in hospitals (20.51% of all cases). The coproculture shows that about 14% of diarrhoeas are of bacterial origin. Thus, our study demonstrates that viral diarrhoeas are as frequent as the bacterial ones. The combination of both has been found in two cases. Our study shows also that a high incidence of rotavirus diarrhoea is found during spring (18.18%). The latex technique is 18.51% less sensitive than ELISA; however, it has the advantages of simplicity, rapidity and low cost. PMID- 2562260 TI - [Histopathological observation of periodontal tissue by tipping movement toward the hydroxyapatite-packed socket]. AB - Tipping movement in orthodontic therapy, which is often performed even in the absence of the adjacent tooth, is represented by the movement following tooth extraction. Such tipping movement in the absence of the adjacent tooth depends not only upon the magnitude of orthodontic force as classified into Stoner 4D, but also upon the condition of the extracted wounds. To evaluate the clinical applicability of HAp-packing used in combination with tipping movement, histopathological investigation was performed in the periodontal tissue with tipping movement toward the adjacent to HAp-packed socket. HAp packing into the extracted wound cavity produced no significant changes in quantitative tipping movement compared with non-HAp packing. HAp packing into the extracted wound cavity, followed by tipping movement, produced compressed sides not only for the moved tooth but also for the anchor tooth. Resorption and disappearance of alveolar septum were observed on the compressed sides of both these teeth for all experimental sides. HAp in an amount enough to meet the narrowing of the extracted wound cavity, associated with tipping movement was discharged from the suture surface. In the same region, fibrous connective tissue lay between the dental root and the HAp mass, which were not adjacent to each other. The fibrous connective tissue presented temporal bone regeneration so as to surround the tooth. Apart from the moving distance, the tissue change showed no essential difference either for a weak force or for a strong force. The above results revealed the usefulness of HAp packing into the extracted wound cavity for tipping movement of the adjacent tooth in the direction of the wound cavity and immediate move ability after extraction. PMID- 2562261 TI - [Usefulness of the bodily movement of the tooth into the hydroxyapatite-packed extracted cavity]. AB - Early application of bodily movement of teeth at an unfinished stage of the healing of the extracted wound i.e. ossification is presumed to increase the risk of jiggling. Using adult dogs, an experimental study was made on bodily movement in the case of HAp packing into the extracted wound cavity to avoid jiggling. When tooth movement was performed immediately after extraction and HAp packing, no encapsulation of HAp with fibrous connective tissue was noted. Either for a weak force or for a strong force, the moved tooth lacked alveolar septum on its compressed side, its dental root being in direct contact with the HAp mass. Remarkable enlargement of periodontal cavity on the tension side and strong osteoclastic resorption of proper alveolar bone were noted. When tooth movement was clone one week after extraction and HAp packing, Either for a weak force or for a strong force, the extracted wound surface was sealed and the HAp mass in the extracted wound cavity got encapsulated by fibrous connective tissue. Alveolar septum has disappeared completely on the compressed side of the moved tooth. Fibrous connective tissue lay between the dental root and the HAp mass and presented its partial osteogenesis. Vigorous bone apposition was noted in the proper alveolar bone surface on tension side. When tooth movement was done one month after extraction and HAp packing, definite ossification was noted in fibrous connective tissue which encapsulated the HAp mass in the extracted wound cavity. Alveolar septum on the compressed side of the moved bone presented resorption and disappearance. For a weak force, gradual ossification was noted in fibrous connective tissue which lay in the same region, and for a strong force, osseous adhesion to the HAp mass was noted. On the tension side, bone apposition was remarkable in proper alveolar bone surface either for a weak force or for a strong force. The above findings revealed thus most suitable time to begin tooth movement was one week after extraction and HAp packing. PMID- 2562262 TI - [Studies on physico-chemical properties of self-setting apatite cement]. AB - A new dental cement was developed by use of tetracalcium phosphate monoxide (Te CP) and dicalcium phosphate dihydrate (DCPD). When an equimolar mixture of the two calcium phosphates was mixed with diluted phosphoric acid, it hardened into a phase of hydroxyapatite (HAp) resembling the main inorganic phase of hard tissues. The present study was undertaken to investigate physico-chemical properties of this newly developed apatite cement and to evaluate its potential as dental cement to clinical application. The setting time was reduced in the presence of synthetic HAp, indicating added HAp to be an accelerator in the setting reaction. The pH of paste rapidly increased to a neutral range from initial solution acidity of about 2.0 and the subsequent setting reaction proceeded in a neutral and weak alkaline pH range. The hardened solid 4 hours after saturation was identified as single phase of apatite by X-ray diffraction. The HAp crystallinity of the set cement varied with the crystallinity of HAp added as setting accelerator. The wet compressive strength of the cement stored for one day in synthetic saliva at 37 degrees C increased to 400 kg/cm2 as added HAp crystallinity decreased. Moreover no disintegration took place when the cement was stored at 37 degrees C in synthetic saliva, which was undersaturated with respect to DCPD but well supersaturated for HAp. In clinical application to animals, no significant cleavage was observed between the hard tissues and the cement by SEM even three months after the cement paste was filled in the tooth cavity of monkeys. In the specimen prepared one week after implantation in the medullary canal of rats, no inflammatory cell appeared. The specimen prepared one month after implantation showed that the set cement was tightly contacted with newly formed bone. These findings strongly suggest that the newly developed self setting apatite cement is useful as a pulp capping agents, root canal fillings and bone substitute. PMID- 2562263 TI - [ESR study on self-curing resin for rebasing during polymerization]. AB - The effects of temperature, concentrations of activator (N, N-dimethyl-p toluidine) and inhibitor (hydroquinone), atmosphere, liquid powder ratio and mixing time on polymerization process of self-curing resin for rebasing were studied by electron spin resonance. When powder and liquid were mixed at a ratio commercially recommended at a physiologic temperature of 37 degrees C, the propagating radical was first detected 5.5 minutes later, of which time interval corresponds to induction time for polymerization. After the induction time, the concentration of propagating radicals increased rapidly with time and reached the maximum level at 27 minutes. The radicals thus formed remained stable for more than 2.5 days when stored in air at 37 degrees C, though their concentration diminished gradually. With increasing temperature for curing, the maximum level of the radical concentration decreased and the propagating radicals disappeared more rapidly, indicating recombination reactions to be perferentially occurring at higher temperatures. Increase in the amount of activator of N,N-dimethyl-p toluidine resulted in increase in the radical concentration. When powder was mixed with a liquid containing less inhibitor of hydroquinone, the radicals were found to remain stable longer. In air, propagating radical was detected faster and free radical was concentrated more rapidly than in the water and nitrogen gas. The fluidity of self-cured resin for rebasing as measured by use of spin probe TEMPO disappeared rapidly with increasing temperature for curing, decreasing the amount of liquid and shortening the mixing time. When curing temperature was raised to 37 degrees C from 22 degrees C at minutes after mixing, the fluidity disappeared 2.5 minutes later, i.e., at 5.5 minutes after mixing. To investigate the effects of the MMA-derivative radicals on growth of HeLa cells, HeLa cells were cultured on resins containing MMA-derivative radicals. The number of HeLa cells on resins containing MMA-derivative radicals was 7/10 of that of HeLa cells on resins containing no radicals. PMID- 2562264 TI - [Effects of endotoxins from periodontopathic bacteria on the collagen metabolism of cultured normal human gingival fibroblasts]. AB - The present study was performed to investigate the effects of endotoxins from periodontopathic bacteria on collagen metabolism. Endotoxins were extracted from Bacteroides gingivalis 381 and Bacteroides intermedius ATCC 25611 using the hot phenol method. A commercially available endotoxin from Escherichia coli 0111: B4 was also used as a control. Human gingival fibroblasts (Gin-1, ATCC CRL 1292) were maintained with DMEM containing 10% FBS. When the fibroblasts became confluent they were exposed to each of the endotoxins in various concentrations (0, 5, 10, 15, 20 micrograms/ml). The effects of these endotoxins on the collagen metabolism of the fibroblasts were assessed on the basis of collagen synthesis and collagenase activity. The former was assessed by 3H-proline incorporation and bacterial collagenase digestable protein. The latter was assessed by fibril assay. The results were as follows: There was no change in glucose consumption, cell viability or morphology under the light microscope when the concentration of endotoxin was 20 micrograms/ml or less. 3H-proline incorporation into protein and collagen synthesis were inclined to decrease. Endotoxin from B. gingivalis resulted in an acceleration of collagenase activity. There findings indicate that the endotoxins from these bacteria might affect collagen metabolism early in gingivitis in vivo. PMID- 2562265 TI - [Basic study of the application of 2 piece-type hydroxyapatite anchor in periodontal therapy]. AB - Clinico-histological studies were made to obtain basic data on the application of 2 piece-type hydroxyapatite (hereinafter referred to as HAP) originally designed and prepared so as to be shaped to make its clinical application easy as an anchor (HAP anchor) in the corrective movement of teeth. An immobility identification experiment and intrusive experiment were performed in order to identify absent loss of anchor to the corrective force and to identify actually practicable tooth movement by means of the HAP anchor, respectively, and clinico histological evaluation was made of its function in both of the above cases. The results revealed no change in the distance of HAP anchor in the former experiment and tooth intrusion of 3.7 mm on average in the latter experiment and, histologically, close surrounding of almost the entire region embedded by the HAP anchor, with newly formed osseous tissues in both experiments. The above results suggest the usefulness of this trial HAP anchor as an anchor to tooth movement in periodontal therapy. PMID- 2562266 TI - [Effect of periodontal regeneration using collagen-coated synthetic bone implant materials. Histopathological study]. AB - Granulated artificial bone implant materials such as hydroxyapatite (HAP) and tricalcium phosphate (TCP) have been frequently used in attempts to restore periodontal tissue lost as a result periodontitis. However, these materials are considered insufficient for the maintenance and stability of granules at sites losing bone, for active bone formation, or for periodontal ligament regeneration. We have now developed a complex consisting of HAP or TCP coated with atelocollagen (which has recently received attention as a biomaterial) and have conducted experiments to determine the effects of this material on the reconstruction of periodontal tissue. Implantations were performed using a HAP atelocollagen complex, TCP-atelocollagen complex, HAP and TCP at three-wall bone defect sites in experimentally-induced periodontitis in dogs. A control group without the implants was included in the study. Histopathological observations were conducted 2, 4 and 12 weeks after surgery. Compared with the control group, the groups implanted with the complex displayed enhanced maintenance and stability of granules, suppression of epithelial downgrowth, and acceleration of new bone and cementum formation. These results indicate that the implant method using an atelocollagen-coated artificial bone implant material would be useful for periodontal surgical treatment, eliminating some of the disadvantages of conventional implant methods. PMID- 2562267 TI - [Bone inductive activity of hydroxyapatite-bone morphogenetic protein complex]. AB - Bone morphogenetic protein (BMP) is known to be a protein which induces new bone at heterotopic sites. Purification of BMP has not been perfected, and obtaining large amounts of BMP is very difficult, so it seems better to use some carrier or frame material for BMP to work effectively. Various kinds of hydroxyapatite (HAP) have been used to repair periodontal osseous defects, but they do not have osteogenetic or osteoinductive properties. If osteoinductive proteins such as BMP could retain their biologic properties after being implanted into living tissue with HAP, it would be an advantage in repairing periodontal osseous defects. In this experiment, we prepared BMP-HAP complex and investigated its osteoinductive activity. BMP was extracted from bovine cortical bones in accordance with the Urist's procedure. The ability of this BMP to stimulate new bone growth was ensured by implantation in the muscle pouch of mice. HAP was synthesized by the wet method. The BMP-HAP complex was implanted in the muscle pouch of mice, and osteoinduction was examined 3, 7, 14, and 21 days after implantation to assess its osteo-inductive ability. New bone formation was studied by roentgenographic and histologic observation. In the BMP-HAP group, new bone formation was seen on the roentgenograms and new cartilage and bone were observed histologically in the tissue surrounding the apatite. In the HAP group, no new cartilage or bone formation was noted. PMID- 2562268 TI - [Clinical evaluation of hydroxyapatite implants in human periodontal osseous defects. Clinical observations over a one-year period following surgery]. AB - The purpose of this study was to evaluate the effect of implants of granular hydroxyapatite (HAP). HAP was implanted into twenty-five vertical bone defects of twenty-one patients as bone graft material. Various clinical and radiographic examinations were performed postoperatively over a twelve month period. Redness and swelling of the gingiva, gingival bleeding, postoperative pain and increased tooth mobility developed transiently, but they all recovered in time. Open wounds and out-flow of HAP disappeared within the first month. After twelve months, mean probing depth decrease was 3.7 mm and clinical attachment gain was 2.5 mm. In all cases there was radiographic evidence of alveolar bone repair. These results suggest that HAP is clinically effective as a bone graft in periodontal therapy. PMID- 2562269 TI - [Clinical study of the anti-calculus effect of a dentifrice containing sodium polyphosphate]. AB - A double-blind clinical study was conducted to evaluate the effect of a tartar control dentifrice containing 1% sodium polyphosphate as tartar control agent compared with dentifrice A containing 5% sodium pyrophosphate and a placebo dentifrice. One hundred forty-eights who continued to have a degree of calculus formation after using a regular dentifrice containing no tartar control agent for 1 month in the pretest, were stratified randomly on the basis of pretest, calculus score, age and sex into 3 homogeneous groups. All subjects who received initial prophylaxis were provided the assigned dentifrice and toothbrush. No instructions regarding frequency or method of toothbrushing were allowed. Assessment of supragingival calculus was made at 4 and 12 weeks using the calculus scoring procedure proposed by Volpe et al. Side effects such as oral irritation, ablation of oral mucosa and discoloration of teeth caused by dentifrices were also diagnosed after 12 weeks of use. The following results were obtained in this study. 1. The test dentifrice had reduced (P less than 0.01) supra-gingival calculus significantly more than dentifrice A and the placebo at 12 weeks. 2. A significant reduction (P less than 0.01) was observed when dentifrice A was compared with the placebo dentifrice. 30.1% reduction was obtained using the test dentifrice and 9.0% reduction using dentifrice A when assessed in subjects who had a pretest VMI score of more than 6.0. 3. No side effects caused by the dentifrice which contained sodium polyphosphate were observed. PMID- 2562270 TI - [Histological study on gingival tissue reattachment after periodontal surgery. Effects of hydroxyapatite ceramic implantation to denuded dentin surfaces in rats]. AB - The purpose of this study was to evaluate histological regeneration of epithelial and connective tissue attachment to the denuded dentin surface after hydroxyapatite ceramic granule (HAP) implantation. HAP (particle size 100-300 microns, burn temperature 900 degrees C) were implanted into dehiscent defects of alveolar bone which eliminated about a 1 x 1 mm area at mesial and palatal sites of the upper 1st molar in 20 male Wistar rats. Subsequently the root cementum was removed totally and dentin was denuded. Same osseous defects were formed but HAP was not implanted at any contralateral site for the purpose of control. Results of 8-week examination using light and electron microscopy were as follows: 1. In the HAP group junctional epithelium was regenerated on the denuded dentin surface 2 weeks after implantation and epithelial down growth was less than at the control site. 2. In the HAP group connective tissue reattachment with new cementum formation was observed at 2 to 4 weeks after implantation, the healing period tended to be shorter than at the control site and the connective tissue reattachment level was located more coronally. 3. In ultrastructural findings collagen fiber bundles attached to the denuded dentin surface had fibers continuing around HAP granules. PMID- 2562271 TI - [Study on hybrid materials compounded with porous hydroxyapatite and culture cells. 1. Attachment and the invasion of cultured cells to porous hydroxyapatite]. AB - The purpose of this study was to investigate the effects of hybrid material compounded with periodontal ligament cells on human teeth and porous hydroxyapatite. As the first step, we tried to produce hybrid materials compounded with periodontal ligament cells and porous hydroxyapatite, and to observe the attachment and invasion of periodontal ligament cells porous hydroxyapatite. Human periodontal ligament cells which had been scraped off the root with a disposable blade were cultured at 37 degrees C in alpha-MEM plus 10% fetal calf serum. The culture cells were attached to the porous hydroxyapatite in vitro. After 3 and 14 days of incubation, the specimens were fixed in gluteraldehyde and examined using SEM. Other specimens were embedded in paraffin and successive specimens were prepared for to remodelling of the attachment and invasion. Many cells attached to the P-HAP granules, and some cells invaded the P HAP granules. The results suggest that when these hybrid materials are implanted in bone defects, the cells remain in the defect for a certain period, and exhibit the functions and characteristics of itself. PMID- 2562272 TI - [Application of CaO-P2O5-MgO-SiO2-CaF system glass ceramics to periodontal therapy. Histopathological observation after implantation in furcation bony defect in monkeys]. AB - This study was carried out in order to determine the efficacy of CaO-P2O5-MgO SiO2-CaF system glass ceramics, which are made as implant materials, for treatment of furcation lesions. Glass ceramic granules were implanted in artificial class II furcation bony defects in monkeys. As controls, non-implanted sites were preserved. Radiographic and various clinical examinations were performed before surgery and at 0, 2, 4 and 8 weeks after implantation. Two, 4 and 8 weeks after implantation, two monkeys were sacrificed and the mandibles were sectioned for histopathological observation. The results obtained were as follows; 1. During the experiment, no clinical problems or abnormal response at the sites of glass ceramic implantation were observed. 2. In clinical observation, no remarkable differences were obtained between control sites and implanted sites. 3. Two and 4 weeks after surgery, remarkable regeneration of bone was shown in the implanted sites. However 8 weeks after surgery, the difference between implanted and control sites was not so clear. PMID- 2562273 TI - [Clinical and histopathological observation of CaO-P2O5-MgO-SiO2-CaF system glass ceramic tooth implants in monkeys]. AB - The present study was undertaken to determine the biocompatibility of a newly developed CaO-P2O5-MgO-SiO2-CaF system glass ceramic tooth implant. Two adult male monkeys were selected and 12 weeks after extraction of M1 and P1, 8 glass ceramic teeth were implanted into alveolar bone. At 1 week after implantation, 6 teeth were allowed occlusal function, and 2 teeth were left free from occlusion as control teeth. The implants were observed for 4-12 weeks and examined histopathologically. The results were as follows: 1. Seven implanted teeth were well maintained clinically, and only one tooth was lost after 5 weeks. 2. In histopathological observation, implanted teeth were surrounded by bone, and connected by bonyankylosis. At cervix of dental implant, connective tissue as also attached firmly to implanted tooth surfaces and epithelial attachment was observed. 3. Although these implants were allowed occlusal function at an early stage (1 week after implantation), osteogenesis around implants was not disturbed. These results suggest that the new glass ceramic implant has good biocompatibility and is useful as an implant tooth. PMID- 2562274 TI - [Steady-state calcium accumulation and its reduction by caffeine in sarcoplasmic reticulum from masseter muscle]. AB - The passive Ca2+ efflux pathways in skeletal sarcoplasmic reticulum (SR) vesicles include the efflux through a Ca2+ channel and a passive leak parallel to the channel and the pump. It is known that caffeine stimulates Ca(2+)-induced Ca2+ release. To gain further insight into the effect of caffeine on Ca2+ flux behavior of canine masseter muscle SR vesicles, the present study focuses on the interaction of steady-state Ca2+ accumulation and passive Ca2+ permeability in the presence or absence of endogenous calmodulin (CaM), which is known to regulate Ca2+ release channel. Caffeine (1) produced ruthenium red- or endogenous CaM-inhibitable reduction of oxalate-supported Ca2+ uptake velocity with no effect on Ca2+, Mg(2+)-ATPase activity; (2) reduced steady-state Ca2+ uptake; and (3) had no effect on the permeability of the SR vesicles to Ca2+, determined by measuring net efflux of Ca2+ after stopping pump mediated fluxes, suggesting that passive Ca2+ permeability is unimportant pathway for changing steady-state Ca2+ accumulation. The inhibitory effect of caffeine on steady-state Ca2+ uptake was moderately abolished by the removal of endogenous CaM from SR vesicles. Inositol 1,4,5-trisphosphate (IP3) caused the same effect as that of caffeine on oxalate supported Ca2+ uptake velocity, steady-state Ca2+ uptake and passive Ca2+ permeability. In summary, the data reveal that caffeine (1) inhibits oxalate entry pathway via inhibition of CaM, and (2) directly modifies CaM-dependent component of Ca2+ fluxes in the SR and reduces steady-state Ca2+ accumulation due to increased Ca2+ release through a Ca2+ efflux pathway which is inhibited by CaM but not due to reduced catalytic activity of the pump; and that the masseter muscle SR vesicles include IP3-sensitive Ca2+ release channel. PMID- 2562275 TI - [Phosphoproteins biosynthesis induced by 1,25(OH)2D3 in the rat calvarial osteoblasts]. AB - The present study attempts to explore the newly synthesized phosphoproteins secreted by osteoblast-like cells incubated with 1,25(OH)2D3. The phosphoproteins, which are non-collagenous proteins, may possess the ability to regulate bone mineral solubility. An osteoblast-enriched cell population isolated from 2 day-old rat calvaria by sequential enzymatic digestion was cultured in a defined medium containing dialized fetal calf serum protein (FCSP, 2 mg/ml) with 1, 5 and 10 x 10(-9)M 1,25(OH)2D3. At confluence, 32Pi (Na2H32PO4, NEX-011) was added for 24 hr. The medium proteins were precipitated by cold 10% TCA, dissolved in 15 mM Tris-HCl, pH 7.4 containing 7 M urea and chromatographed on hydroxyapatite columns (Bio-Rad, HTP). After stepwise elution with 6 mM, 0.1, 0.5 and 1.5 M Pi Hepes buffer pH 7.4 containing 3 M urea and 5 mM levamisole, the phosphoproteins were applied to 10% SDS-PAGE and autoradiographed. The 32Pi incorporated phosphoproteins of 75K, 66K, 58K, 42K, 38K, 24K, 22K, 19K, 15.5K, 13K and 3.5-10K molecular weight which were bound on a hydroxyapatite column were identified on autoradiograms of SDS-PAGE. The synthesis of 19K phosphoprotein was stable. However the synthesis of 75K, 66K, 38K and 15.5K phosphoproteins were increased by 1,25(OH)2D3. Therefore, 1,25(OH)2D3 induced phosphoproteins synthesized in rat calvarial osteoblasts, which can bind tightly on hydroxyapatite, may regulate the solubility of bone mineral and play a role in maintaining a blood/bone equilibrium. PMID- 2562276 TI - [Effect of compound 48/80 on masseter muscle sarcoplasmic reticulum calcium transport system]. AB - To define the role of calmodulin in Ca2+ fluxes behavior of canine masseter muscle sarcoplasmic reticulum (SR) vesicles, the effect of condensation product of N-methyl-p-methoxy-phenethylamine with formaldehyde (compound 48/80), a selective and powerful inhibitor of calmodulin-regulated function, on Ca(2+) ATPase activity, oxalate-supported Ca2+ uptake velocity, and on interaction with Ca2+ permeability and Ca2+ loading at steady-state were evaluated. Compound 48/80, at concentrations of 10 to 100 micrograms/ml, reduced oxalate-supported Ca2+ uptake velocity without affecting Ca(2+)-ATPase activity. In the presence of 10 micrograms/ml compound 48/80, there was a shift of pH- or temperature-response curve of oxalate-supported Ca2+ uptake velocity, but not of Ca(2+)-ATPase activity, down. It was found that Arrhenius plots of the Ca(2+)-ATPase activity show a break at about 21 degrees C in the presence or absence of 10 micrograms/ml compound 48/80, and that compound 48/80 has no effect on Arrhenius plots of the oxalate-supported Ca2+ uptake velocity. Furthermore, Ca2+ loading at steady state, but not passive Ca2+ permeability, was decreased by compound 48/80 at low concentrations (1-2 micrograms/ml). The results of this study suggest that calmodulin-dependent process plays a functional role in the coupling of ATP hydrolysis and Ca2+ accumulation, perhaps through regulation of Ca2+ release channels in masseter muscle SR membrane. Calmodulin-dependent component of Ca2+ fluxes in the SR vesicles may be directly modified by compound 48/80, thereby diminishing Ca2+ accumulation without affecting the Ca2+ uptake mechanism. PMID- 2562277 TI - [Studies on the relation between glucose metabolism and c-AMP formation in dental pulps in the presence of inflammatory chemical mediators in vitro]. AB - The relationship between glucose metabolism and cyclic-AMP production in dental pulp in the presence of chemical mediators was investigated in vitro. It is generally accepted that oxidation of glucose-6-14C is indicative of metabolism by the glycolytic pathway whereas that of glucose-1-14C occurs by the hexose monophosphate shunt. The 14CO2 productions from both routes were compared in dental pulp from cattle and rats in the presence of each of several chemical mediators: bradykinin (1.7-3.3 micrograms/ml), prostaglandin E1 (0.3 micrograms/ml), prostaglandin E2 (0.3 micrograms/ml), histamine (33 micrograms/ml), and 5-hydroxytryptamine (33 micrograms/ml). The effects of dental filling materials on glucose oxidation, and cyclic-AMP production by chemical mediators in pulp tissues were also investigated. The results obtained were as follows: 1) Glucose oxidation in dental pulp was stimulated by chemical mediators generally by way of the Embden-Meyerhof Parnas pathway, and was further stimulated by the medium containing bradykinin. However, it was depressed in the presence of higher concentrations of chemical mediators, especially depressed in the HMS pathway. 2) The oxidation ratio of glucose-1-14C to glucose-6-14C (G1/G6) in dental pulp was 4 to 8 in the cattle and 0.6 in the rat, showing clear differences in glucose oxidation between the two animals. 3) Moreover, glucose oxidation in rat dental pulp was 60 to 80 times higher in the EMP pathway and 5 to 10 times higher in the HMS pathway than those in the cattle. 4) Dental filling materials such as silicate cement, zinc phosphate cement, calcium hydroxide, and eugenol cement severely depressed glucose-6-14C oxidation in bovine dental pulp when used at high concentrations, but not at low concentrations. 5) The chemical mediators tested in this experiment (PGE1, PGE2, histamine, 5-HT, bradykinin, and substance P) stimulated cyclic AMP production in rat dental pulp. The production was highest with PGE1 and PGE2. The cyclic AMP production was further stimulated by addition of histamine or 5-HT to the medium containing PGE1 or PGE2. PMID- 2562278 TI - [Study of hard mineralized tissues effect of calcium-deficient diets with and without cholecalciferol supplementation on the incisor dentine of rats]. AB - It has been known that both tooth and bone have an apatite structure similar to that of mineral hydroxyapatite. The apatite crystal in living tissues of tooth and bone generally is constructed from submicrocrystals and has many impurities. It seems that chemical and physical aspects of resistance of hard tissues depend on the diet. Consequently, the matrix of organic, free radicals in X-ray irradiated hard tissues were studied by means of electron spin resonance (ESR). The effects of calcium-deficient diets with and without cholecalciferol supplementation on the incisor dentine of rats were examined by the rate of decay of organic, free fadicals in X-ray irradiated incisor dentine of rats. The crystal size of incisor dentine became smaller in case of calcium contained diet. PMID- 2562279 TI - [Practical application of diamond points in mucosalplasty of edentulous ridges]. AB - In order to establish a new operative method of mucosalplasty in the edentulous ridges to improve pontic adaptation, newly revised diamond points were produced and tested. The efficiency of these diamond points were tested on the edentulous ridges of dogs in relation to its shape, size of the diamond particle, and appropriate rotational speed, and the tissue damages and healing processes were observed under a light microscope and SEM. The results were as follows: 1. Light microscopic observations showed immediate post operative necrosis of the wound surface, and degeneration below the necrosed layer of high rotational speed diamond points. 2. Tissue damage became larger as the size of the diamond particle increased, but there appeared to be no influence upon the healing process of the wound. 3. Tissue damage also became larger as the diameter of the diamond point increased, but again no influence upon the healing process was observed. 4. Middle to low rotational speeds showed no differences in tissue damage or wound healing, but high rotation caused larger tissue damage and prolongation in the healing process. 5. The most suitable condition for mucosalplasty revealed the size of the diamond particle to be 200 microns, and rotational speed to be between 12,000-20,000 rpm. PMID- 2562280 TI - [Prosthodontic analysis in mandibular ridge augmentation with hydroxyapatite particle. 2. Evaluation of masticatory function and overall assessment]. AB - Ten patients received augmentation of deficient mandibular alveolar ridges using particulate hydroxyapatite (HAP). Masticatory ability was evaluated preoperatively and postoperatively by a questionnaire with 35 foods listed and by the spectrophotometric method using ATP granules. The results showed that masticatory ability when wearing newly constructed complete dentures was significantly enhanced compared to that when wearing old dentures or relined dentures, and that the surgical procedure and the use of the surgical stent greatly improved masticatory ability when wearing the new dentures. The sensitivity of the mucose membrane of the augmented mandibular alveolar ridge was examined during a postoperative period of 24 months by using an electric nerve stimulator. The sensitivity of the mucose membrane decreased postoperatively, especially at the buccal site of the molar region, the so-called buccal shelf. The level of sensitivity increased periodically, and did not return to the average level for normal edentulous mucosa even 24 months after the augmentation. The patients were highly satisfied with the new dentures and the HAP augmentation judging from the questionnaire. From these points of view, HAP augmentation in edentulous patients having severely resorbed alveolar ridges has proved to be beneficial in allowing the recovery of masticatory function. PMID- 2562281 TI - Gestational trophoblastic tumors and malignant ovarian germ cell tumors. PMID- 2562282 TI - Association of human papillomavirus with gynecologic cancer. PMID- 2562283 TI - Gynecologic cancer. PMID- 2562284 TI - Molecular biology of lymphoid malignancies. PMID- 2562286 TI - Staging and surgical approaches to carcinoma of the breast. PMID- 2562285 TI - Biology of breast cancer and its clinical implications. PMID- 2562287 TI - Radiotherapy in the management of primary and advanced disease. PMID- 2562288 TI - Spectroscopic studies of cobalt-substituted ferredoxin from Clostridium pasteurianum. AB - Ferredoxin from Clostridium pasteurianum substituted with two Co atoms did not give any cobalt EPR signal at 8 K as isolated, but upon reduction with sodium dithionite, a broad signal appeared with g values that indicate highspin (S = 3/2) Co(II). These signals were distinct from Co(II)-dithiothreitol signals, and disappeared upon reoxidation with air. Under anaerobic incubation of apoferredoxin with Co(II), a green derivative showed a visible spectrum typical of tetrahedral Co(II)-thiolate coordination, which shifted dramatically upon exposure to air. The 1H-NMR spectrum of the aerobically isolated protein is reported at 300 MHz; magnetic susceptibility measurements were indicative of a diamagnetic species. These spectroscopic studies indicate that Co(II)-substituted ferredoxin is oxidized to low-spin Co(III)-ferredoxin in the presence of sulfide and oxygen. The diamagnetic Co(III) state could reversibly be reduced to highspin Co(II) by sodium dithionite. PMID- 2562289 TI - Rapid liquid chromatographic analysis of mitoxantrone in plasma with C18 sample purification. AB - A useful and rapid method for the analysis of mitoxantrone in human plasma is described. We purified the sample with a C18 Sep Pak cartridge pre-treated with methanol, water and 0.05 M ammonium phosphate (pH = 2.75). The drug and internal standard (Methylene Blue) were eluted from the cartridge with 1 M acetic acid in methanol and were separated on a 10 microns particle size CN Resolve cartridge in conjunction with a radial compression liquid chromatograph. The mobile phase consisted of a mixture of 0.05 M ammonium phosphate, acetonitrile and methanol (60:35:5, by vol), and the detection was performed spectrophotometrically at 660 nm. The peak height ratio (drug/internal standard) varied linearly (r greater than 0.993) with concentration over the range examined 0.01-3 micrograms ml-1, and the inter- and intra-run precision at high, medium and low concentrations were good; CV ranged from 2.52 to 7.2%. There was no interference from other anticancer drugs or analgesics. We applied the described method to investigate the pharmacokinetics of mitoxantrone using a rabbit as an in vivo model. PMID- 2562290 TI - Determination of the optical purity of indacrinone by proton nuclear magnetic resonance spectroscopy using chiral lanthanide chelates. AB - A simple, specific and reliable 1H-NMR spectroscopic method for the quantitative determination of the optical purity of indacrinone is described. After conversion of the S(+)- and R(-)-enantiomers to their methyl ester derivatives, they were coordinated with chiral europium(III) or praseodymium(III) shift reagents in CCl4/C2HCl3 (2:1). The optical purity was calculated from the relative intensities of the enantiomeric resonance signals for the protons of the methyl groups at position C(2) of the indanone ring. Mean +/- SD (n = 6) recoveries of S(+)-indacrinone from synthetic enantiomeric mixtures amounted to 99.75 +/- 0.63% when using europium(III) and 100.01 +/- 0.55% when using praseodymium(III). PMID- 2562291 TI - FIA-extraction applied to the limit test for heavy metals. AB - A method is presented that allows rapid determination of the total concentration of heavy metals in a sample. The method is based on FIA-extraction and photometric measurement of the metals as their dithiocarbamate complexes. The analytical parameters have been chosen such that the sensitivities for toxic elements are enhanced compared with those of less toxic heavy metals. The sampling capacity of the system is 40 samples h-1 and the repeatability (RSD) is 1.9% at 0.1 mg 1(-1). Raw materials for the production of pharmaceuticals as well as analytical grade salts and household commodities have been tested. PMID- 2562292 TI - [Abrikossoff's tumor (AT)]. PMID- 2562293 TI - Non-ligand-activated chloride channels of skeletal muscle and epithelia. PMID- 2562294 TI - [Characteristics and role of the gluteal herpes in a female population]. AB - Based on the fact that the gluteal herpes may constitute the clinical expression of the reactivation of a Herpes simplex virus latent at the sacral lymph node, we investigated a group of women who were carriers of gluteal herpetic infection, the characteristics of the infection, the virus type principally associated to it, and its possible relation with the genital herpes. Forty one women with gluteal herpes verified by virologic laboratory were studied. 75.7% of these women had had in addition to this herpetic infection in other places, mainly genital, with an average of 7.2 of recurrent episodes per year, (range: 1 to 18 episodes yearly). 78% of the isolated virus was typified as HSV-2 by the use of monoclonal antibodies. It is stand out the importance of considering the background of gluteal herpes as causative of classification of herpetic high risk. PMID- 2562295 TI - [Midazolam in anesthesiology]. AB - Midazolam (MDZ) (8-chloro-6(2-fluoropheny 1)-1-methyl-4H-imidazol-[1,5a] [1,4]benzodiazepine) is an "annelated" benzodiazepine (BZDs) synthetized in 1976, characterized differently with the "classical" BZDs by a five-membered heterocycle fused on position 1,2 of the diazepine nucleus. This fused imidazol ring modifies the properties inherent in the "classical" BZDs in at the least three aspects: solubility, metabolisation and the stability in aqueous solution. MDZ, having similar properties with the "classical" benzodiazepines, has better local tolerance, faster onset of action, greater plasmatic clearance, shorter half-life elimination (1.7-2.4 hr) with no active metabolites. With a bioavailability of 92% (IV), 82-91% (IM) and 50-52% "per os", the CNS effects of MDZ are similar in all three ways. In anesthesiology we can administer MDZ as an anesthetic premedication ("per os", IM or IV), anesthetic induction and maintenance and IV sedation in locorregional anesthetic procedures or diagnostic and therapeutic explorations. MDZ has a great safety margin, moderate respiratory and cardiovascular effects and lacks of teratogenic or embryotoxic effects. PMID- 2562296 TI - The role of cytophotometric and autoradiographic investigation for the prolongation of the survival of patients with astroglial tumours. PMID- 2562297 TI - Recent aspects of erythropoietin and its clinical utility in the future. PMID- 2562298 TI - Modulation of striatal serotonin metabolism by baclofen, a gamma-aminobutyric acidB receptor agonist. AB - Intraperitoneal (i.p.) administration of racemic baclofen (10-40 mg/kg), a gamma aminobutyric acidB (GABAB) Receptor agonist, increased striatal and hippocampal 5 hydroxytryptophan (5-HTP) accumulation after inhibition of L-aromatic amino acid decarboxylase. The baclofen-induced increment of 5-HTP accumulation in the striatum showed a much greater magnitude and longer duration than that in the hippocampus. In contrast, systemic application of baclofen (10-40 mg/kg, i.p.) failed to modify the rate of serotonin (5-HT) disappearance during inhibition of tryptophan hydroxylase in the striatum and hippocampus. Acute cerebral hemitransection decreased striatal 5-HTP accumulation and completely blocked the ability of baclofen to enhance 5-HT synthesis in the striatum. Furthermore, there were no changes in striatal 5-HTP accumulation after intrastriatal infusion of baclofen and GABA. These findings suggested that systemically applied baclofen facilitates in vivo 5-HT synthesis in the striatum via primarily stimulating GABAB receptors located in the extra-striatal area(s). PMID- 2562299 TI - Interferon release by LPS-treated macrophages from patients affected by neoplasia. AB - Concerning the modulatory role of LPS on immunocompetent cells, the production of interferon by macrophages primed with human gamma interferon (HUIFN gamma), was studied. In particular the production of IFN from primed macrophages of patients affected by breast cancer, differentiated in presence of autologous serum or in serum from healthy donors, was compared with the IFN production from macrophages of healthy donors. The levels of endotoxin-induced IFN were enhanced by priming macrophages with HUIFN gamma. The "in vitro" treatment with Escherichia coli LPS restores the interferon production of primed macrophages, obtained from patients affected by breast cancer, differentiated in presence of serum from healthy donors. Moreover, LPS treatment of primed macrophages from patients, differentiated in autologous serum, did not influence the interferon production of these cells. Nevertheless, primed macrophages from healthy donors appeared to be more sensitive to LPS treatment in comparison with primed macrophages from patients affected by breast cancer. PMID- 2562300 TI - [Biological activity of gamma interferon on various cell types in human and murine systems]. AB - The effects of interferon (IFN gamma) and of IFN alpha/beta on normal T and B lymphocytes and various T and non-T human and murine cell lines have been investigated. IFN gamma, unlike IFN alpha/beta, did not promote the antiviral state in T cells. The lack of antiviral activity was confirmed at the biochemical level by the finding that 2',5' oligoadenylate synthetase activity is not induced in T cells by IFN gamma only. PMID- 2562301 TI - [Alithiasic cholecystitis and sclerosing cholangitis caused by Cryptosporidium and cytomegalovirus in a patient with HIV infection]. AB - A 23-year-old female, an ex-parenteral drug abuser with HIV infection, developed multiple pericholangitic abscesses secondary to lithiasic cholecystitis and sclerosing cholangitis by Cryptosporidium and cytomegalovirus. We emphasize the clinical clues and the diagnostic role of endoscopic retrograde cholangiopancreatography (ERCP), as well as the contribution of the latter to surgical therapy (transendoscopic papillotomy). We also review the reported cases of biliary diseases by Cryptosporidium and/or cytomegalovirus in the literature. PMID- 2562302 TI - [Hyperthermia caused by exertion in cytomegalovirus infection]. PMID- 2562303 TI - [Healthy carriers of enteropathogenic micro-organisms among the child population of Seville]. AB - We have studied for 1-year period a group of 144 children (31 newborn infants, 62 aged 1 year and 51 aged 2 years) who were randomly selected from the registrar's office of Sevilla with the purpose of determining the incidence of diarrhoea and the prevalence of enteropathogenic microorganisms. Two samples of faeces (one at the beginning and the second by the second semester of the 1-year period) were obtained from all children which were processed for culture and parasite and rotavirus examination. We found a prevalence rate of enteropathogenic Escherichia coli carriers (EPEC) of 7%, of Giardia lamblia of 4% and of rotavirus of 14%. The state of EPEC was more frequent among children from high social-economic status. The state of G. lamblia carrier was six-fold higher in children with body weight alterations and in non-vaccinated or incompletely vaccinated children. PMID- 2562305 TI - [Neurological manifestations of Lyme disease]. PMID- 2562304 TI - [Evaluation of serological criteria for the diagnosis of infectious mononucleosis]. AB - We have evaluated the application for the diagnosis of infectious mononucleosis of several serologic criteria, including measurement of concentrations of IgM against viral capsid antigen (VCA) of Epstein-Bar virus (EBV) by ELISA, detection of heterophil antibodies and the presence of anti-VCA IgG in absence of IgG against nuclear antigen of EBV (EBNA). Such criteria were evaluated by using a board of sera obtained from cases who had clinically been diagnosed of infectious mononucleosis and identified in the laboratory as produced by EBV (84 sera) or cytomegalovirus (CMV) (9 sera). The measurement of anti-VCA IgM by any of the two tested methods is the best criteria for the serologic diagnosis of infectious mononucleosis. Positive anti-VCA IgM reactions were detected by IIF and ELISA in sera of patients with infectious mononucleosis-like syndrome due to cytomegalovirus, so an alternative method to the measurement of anti-VCA IgM is needed, the most adequate is the measurement of anti-VCA IgG in the absence of anti-EBNA IgG. PMID- 2562306 TI - [Acute pericarditis caused by cytomegalovirus in a normal host]. PMID- 2562307 TI - [Pulmonary infections in immunocompromised patients]. PMID- 2562308 TI - [Therapy of infections caused by opportunistic pathogens: viruses and parasites]. PMID- 2562309 TI - Epstein-Barr virus and its antigens. PMID- 2562310 TI - Immunogenetics of murine and feline retroviruses. PMID- 2562311 TI - Immunologic response to influenza virus infection. PMID- 2562312 TI - Retroviral-related genes, proto-oncogenes, and breast cancer. PMID- 2562313 TI - FIA determination of chlorhexidine by means of the precipitation with Cu(II). AB - The determination of chlorhexidine in pharmaceutical formulations is carried out using flow injection analysis (FIA) with measurement by atomic absorption spectrometry (AAS). The method is based on the formation of a copper-biguanide complex precipitate when the sample is injected into an ammoniacal copper solution. The precipitate is retained on a plastic or paper filtering device. A nitric acid stream dissolves the precipitate and carried the Cu(II) to the AAS detector. The chlorhexidine is determined over the range 5-20 ppm. The influence of interfering substances is investigated. PMID- 2562314 TI - Lung tissue binding of iodobenzyl-propanediamine: involvement of beta-adrenergic receptors. AB - The basic compound N-N-N'-trimethyl-N'-(2-hydroxy-3-methyl-5-iodobenzyl)-1,3- propanediamine (HIPDM) accumulates in human and rabbit lungs, where it forms a slowly effluxable pool. In isolated perfused rat lung, HIPDM is taken up by a saturable, energy-independent mechanism, which is competitively inhibited by imipramine, chlorpromazine and propranolol. To ascertain whether beta-adrenergic receptors are involved in the binding process of HIPDM to lung tissue, the ability of unlabelled HIPDM to displace the beta-adrenergic receptor ligand [125I]iodocyanopindolol (ICYP) from rabbit lung beta-receptors was examined. Lung microsomal membrane fractions (75 micrograms ml-1) were incubated at 37 degrees C for 3 h with 68 pM ICYP (with or without 1 microM of (+/-)-propranolol) in the presence of HIPDM (10(-10)-10(-3) M). Bound and free radioactivity were separated through glass-fibre filters and the retained radioactivity was counted in a gamma spectrometer. HIPDM competed with ICYP for beta-adrenoceptors (13% displacement at 10(-5) M. 50% at 5 x 10(-5) M, and 90% at 2 x 10(-4) M). The inhibition curve of ICYP binding by HIPDM was similar to that observed for (-)-noradrenaline. Although the results of the in vitro studies cannot be extrapolated to in vivo conditions, they suggest that beta-adrenergic receptors may be involved in the observed lung uptake of the basic amine HIPDM. PMID- 2562315 TI - Chiral 1,4-benzodiazepin-2-ones: relationship between stereochemistry and pharmacological activity. AB - Pure enantiomers of 3-substituted-1,4-benzodiazepin-2-ones, obtained by HPLC resolution on chiral stationary phases, show significant differences in their pharmacological activity. The occurrence of biotransformation during the pharmacological test is monitored using a new chromatographic method. The reliability of the pharmacological activity data is discussed. PMID- 2562316 TI - [Infections in the newborn infant]. PMID- 2562318 TI - [Papillomavirus genital infections]. PMID- 2562317 TI - [Prevalence of antibodies against Toxoplasma gondii, rubella virus, cytomegalovirus and herpes simplex virus in pregnant women of Catalonia]. AB - A seroepidemiological survey was carried out to evaluate the prevalence of antibodies against toxoplasma, rubella virus, cytomegalovirus (CMV) and herpes simplex virus in pregnant women from Catalonia during 1985. The study was carried out in a representative sample of the pregnant women cared for in public and private hospitals from Catalonia, which was selected in maternal clinics by random sampling. Antibody measurements were carried out with the following techniques: toxoplasma, IFI (positive values greater than or equal to 1:10); rubella virus, Iha (greater than or equal to 1:10); CMV and herpes simplex virus, CF (greater than or equal to 1:10). 50.21% of pregnant women had antibodies against toxoplasma, 97.48% against rubella virus, 73.31% against CMV, and 80.93% against herpes simplex virus. The presence of antibodies against CMV and herpes simplex virus were significantly associated ( less than 0.00001). The demographic and socioeconomic variables associated with the prevalence of antibodies were analyzed, and they were compared with other results from the literature. PMID- 2562319 TI - [Evaluation of 2 commercial monoclonal antibody kits for the identification of herpes simplex virus]. AB - Two commercial monoclonal antibody kits (Virgo and Mikrotrak for the identification of Herpes simplex virus (HSV) have been assessed. In 65 positive HSV cultures from clinical samples, 25 strains (38%) were identified as HSV1 with both kits. The Virgo kit identified 37 strains (57%) as HSV2, and did not identify the remaining 3 (5%). With the Microtrak kit, 34 strains (52%) corresponded to type 2, 2 (3%) strains remained nonidentified, and in the remaining 4 isolates (6%) both viral types were identified. The concordance between the two evaluated kits was complete in 86% of strains. In the five strains not identified by immunofluorescence, genome analysis was carried out with endonucleases: Hind III and Xho I. Two of these isolates were identified as HVS1, another as HSV2, and in the remaining two both viral types were identified. Overall 6 mixed infections (9%) were detected, 4 with the Microtrak kit and endonucleases and 2 with only the latter method. The Microtrak system shows some advantages such as quickness, easy readings and the possibility to diagnose infections where both types of virus participate. PMID- 2562320 TI - [Genital herpes of mixed etiology]. AB - The relevance of mixed genital infections in our area was evaluated, and we report their clinical and epidemiological features for both viral types. We evaluated overall 54 Herpes simplex virus (HSV) isolates from 44 patients seen at the clinic of sexually transmitted diseases (STD) of the Sevilla School of Medicine during 1986 and 1987. The identification of the isolated strains was made by direct immunofluorescence with type-specific monoclonal antibodies. In those isolates where we detected both viral types confirmation was obtained with the endonuclease analysis: Hind III and Xho I. Four mixed infections were diagnosed, representing 7% of all HVS isolates of genital origin. Two were classified as initial nonprimary infections, and the remaining two were asymptomatic infections. Two patients showed simultaneous infections by other organisms. PMID- 2562321 TI - International collaborative study of the retention reproducibility of basic drugs in high-performance liquid chromatography on a silica column with a methanol ammonium nitrate eluent. AB - An international collaborative study between 10 laboratories has been carried out to study the reproducibility of the separation of basic drugs on silica columns. The laboratories used common solutions of drugs on both a common batch of packing material and different batches of the same brand of packing material. These were also compared with separations on other brands of packing material. Variations within-batch, within-brand and between brands have been compared. The retentions of the drugs were compared using retention times, capacity factors and relative capacity factors compared with an internal standard. The last method was found to give the most reproducible results. Considerable variations were found between the different brands of silica with a smaller variation between the batches of a single silica brand. However, unlike earlier studies, significant variations were found for separations on a single batch of silica which were partly attributed to differences in eluent preparation and column temperature. PMID- 2562322 TI - A study of the metabolism of dimethylformamide in the rat by high resolution proton NMR spectroscopy. PMID- 2562323 TI - Use of electron spin resonance spectroscopy of spin labels for studying drug induced membrane perturbation. AB - The use of electron spin resonance spectroscopy of spin labels is reviewed in the context of drug-induced membrane perturbation. The correlation between membrane perturbation and biological effects is also considered. PMID- 2562324 TI - Proton NMR spectroscopy of bile for monitoring the excretion of endogenous and xenobiotic metabolites: application to para-aminophenol. PMID- 2562325 TI - Determination of halide acid salts of organic bases and quaternary ammonium compounds by titration. PMID- 2562326 TI - [Use of the centrifugation-culture technique for the rapid diagnosis of cytomegalovirus]. AB - Cell culture is the reference method for the detection of cytomegalovirus (CMV) in clinical samples. However, owing to the slow development of cytopathic effect (CPE), the cultures should be observed for at least four weeks before being discarded as negative. We have used a monoclonal antibody for the immunofluorescent detection of the early CMV antigen 48 hours after the inoculation of the samples, by means of centrifugation on embryonic human lung fibroblasts. Fifty-nine out of the 566 investigated urine samples were positive for early antigen in culture and 57 in conventional culture on the same cellular type (mean time of development of CPE 11.8 days). A modification facilitating sample processing and a considerable saving of reactive is described. The technique for the detection of early CMV antigen in culture after centrifugation can be used for the diagnosis of CMV infection owing to its high sensitivity and the rapidity of the results. PMID- 2562327 TI - [Prevention of viral infections transmitted by semen in volunteer donors for assisted reproduction. 2 years' experience]. AB - During a 2 years period the serological markers of viral infections were evaluated in the semen of 24 donors who were involved in assisted reproduction techniques. The presence of cytomegalovirus, hepatitis B virus and human immunodeficiency virus were investigated. The results were as expected, no data of viral infection at the time of donation being found because the study population was a highly selected one. Taking into account all the known microorganisms that can be transmitted through semen, a protocol for the prevention of infections in the mother and in the prospective newborn is suggested. PMID- 2562328 TI - [Neural involvement in hanseniasis]. PMID- 2562329 TI - [Acid- and neutro-detergent fiber and minor mineral contents in maize and its tortilla]. AB - The present study was carried out to complete available information on the nutrient contents of corn tortilla particularly mineral elements and acid and neutro-detergent fiber. Results indicated that the tortilla weight should be considered in dietary surveys. Protein and ash contents are slightly higher in tortillas than in maize, in the first case due to carbohydrate losses that occur during the process, and in the case of ash, due to the lime used during the cooking process. The process also induces changes in the acid- and neutro detergent fiber in maize. Similar yields related to the decrease of neutro detergent and the increase of acid-detergent fiber were found in both products. The cell walls content decreases in the tortilla, with an increase in cellular content. With respect to minerals, an increase in calcium content was found, giving a better Ca:P balance, as well as increases in Fe, Cu and Zn. PMID- 2562330 TI - [Intake of dietary fiber in the Central American isthmus: nutritional implications]. AB - Mean intakes of dietary fiber (DF) were estimated in the population of Central America and Panama, using the results of dietary surveys conducted in 1969 and 1986 both in rural and urban areas, as well as data on the DF content of foods as consumed in the region. Data on preschool children were also estimated. The results indicated that DF intake in urban areas is lower than that of rural areas, particularly in Costa Rica and Panama. In 1969, intake varied from 32g in El Salvador to 15g in Panama in urban areas, while in rural areas intake was from 45g in El Salvador to 13g in Panama. The foods which contributed most to the total intake in Guatemala, El Salvador and Honduras were tortillas and beans, while in Costa Rica and Panama, beans provided the largest intake. In preeschool children, intake was 12.5g in El Salvador and 5.4g in Costa Rica in 1969, which is the same tendency as that found for adults. From more recent data on food intake, it was found that DF intake had decreased in the rural areas of El Salvador, Honduras and Costa Rica, being between 4 and 9% in the first two countries and 12% in Costa Rica. In the urban area of the latter, from 1969 to 1986 a decrease in DF intake of around 20% has taken place. PMID- 2562331 TI - [Effect of the seed coat on the hardening of common beans (Phaseolus vulgaris)]. AB - Samples of cotyledons and whole black beans (Phaseolus vulgaris), Tamazulapa variety were stored during six weeks at 37 degrees C and 90% relative humidity to establish chemical and physical changes which occur during storage, and to determine the role played by the seed coat in the hard-to-cook phenomenon. After the storage period, samples of whole beans were divided in two subsamples, with and without the seed coat. These two samples and the cotyledons were analyzed for cooking time, water absorption, dietary fiber, tannic acid, soluble pectins and phytic acid. Cooking time of the whole beans increased from 99 to more than 480 minutes in the six-weeks period; for the cotyledons this value increased from 45 to 111 minutes. Cooking time of the dehulled bean, stored as whole bean, increased from 45 to 111 minutes. Cooking time of the dehulled bean, stored as whole bean, increased from 45 to 103 minutes. Water absorption in the whole beans and the cotyledons decreased, although in the cotyledons it was higher, due perhaps to the great absorption capacity to the seed coat. No changes were observed in the dietary fiber content of the cotyledons nor in the beans dehulled after storage. However, in the whole grains neutro-detergent fiber decreased, while acid detergent fiber, cellulose and lignin did not present significant changes. On the other hand, soluble pectates decreased in the whole bean and in the cotyledons; nevertheless the tannin content (as tannic acid) decreased only in the whole beans (from 3.28 to 1.64 mg/g). The data obtained suggest that the seed coat plays a significant role in the hard-to-cook process of hardening of the bean, before and during storage. PMID- 2562332 TI - Radiolabelled unprocessed bran: validation of a practical labelling technique and evaluation of gastrointestinal transit profiles in health. AB - Using a radiolabelled bran preparation, a large field of view gamma-camera and a computer, a method has been developed to define and analyse the gastrointestinal transit profile of solids. Unprocessed bran, labelled with technetium (99mTc) sulphur colloid is incorporated into biscuits cooked in a microwave oven. With this preparation, less than 3.4% of the isotope dissociated from the bran in a gastric milieu, and more than 90% of recovered radionuclide activity, were retained with the solid phase during small intestinal transit in subjects with stable ileostomy function. A normal range for gastric emptying (GE), small intestinal transit (IT) and colonic filling (CF) was established in 15 healthy volunteers. Anterior and posterior images were taken until all isotope was present in the colon. Using regions of interest for the stomach and colon, decay corrected geometric mean curves for GE and CF were generated. GE was characterized by a lag phase followed by a linear emptying phase. The GE profile (mean and standard deviation) was described by 'power exponential' parameters of half-emptying time (1.6 +/- 0.4 h) and beta (1.5 +/- 0.2). The IT time was 3.9 +/ 1.5 h. The onset of CF was determined visually (4.1 +/- 1.5 h), and the time of 50% CF filling was determined from the colonic filling curve (5.5 +/- 1.4 h). This technique should provide a useful, practical, clinical and research test to study the pathophysiology and pharmacology of intestinal motor disorders. PMID- 2562333 TI - Parietal cells in duodenal ulcer disease: a histochemical study of the effects of omeprazole and ranitidine on mitochondrial activities. AB - H2-Receptor antagonists and omeprazole, a H-K ATPase inhibitor, inhibit acid secretion from the parietal cells. The ultrastructural changes of the parietal cells after treatment have been described, but the changes in the mitochondrial activity which reflect the energetic metabolism were not well defined. To study the effect of omeprazole and H2-receptor antagonists on the mitochondrial activity of the parietal cells, endoscopic biopsies were taken from nine patients with duodenal ulcer before and after treatment with either 10 mg or 20 mg omeprazole each morning, or 150 mg ranitidine twice daily for 2 weeks, given in a double-blind randomized manner. Three patients with healed duodenal ulcer who were on maintainence treatment with 150 mg ranitidine nightly for 1 year had an endoscopy performed after 4 and 12 months and two non-ulcer dyspeptic patients were recruited as controls. Three biopsies were taken during each endoscopy from the body of the stomach. The mitochondrial activity was assessed by the reaction of succinic dehydrogenase with nitroblue tetrazolium and of cytochrome oxidase with naphthoic acid mixed with N-phenyl-p-phenylenediamine, according to the intensity of the staining reaction. After treatment with omeprazole or ranitidine, the mitochondrial activity decreased appreciably and returned to the pretreatment level on cessation of treatment. Patients on maintainence ranitidine showed decreased mitochondrial activities after 4 months, which, however, returned to pretreatment levels in two patients. It is concluded that short-term treatment with omeprazole or ranitidine resulted in reversible suppression of mitochondrial activity while long-term treatment with ranitidine resulted in an initial suppression followed by a tendency to return to pretreatment level despite continued treatment. PMID- 2562334 TI - Role and nature of plasma membrane carrier proteins in the hepatic transport of organic anions. PMID- 2562335 TI - [Epidemiology of herpes simplex virus y papilloma virus: current concepts]. PMID- 2562336 TI - Pulp capping with hydroxyapatite ceramic in rat molars. AB - Direct pulp capping with a hydroxyapatite ceramic in rat molars induced the formation of reparative dentin in the exposure sites after 2 weeks. In addition, foci of acute and chronic pulpitis were detected. After 6-8 weeks of application of the material partial or total necrosis occurred. The use of hydroxyapatite as a possible pulp capping material is discussed. PMID- 2562337 TI - Oral cavity herpes simplex virus--a risk factor to dental personnel and patients. An overview. AB - Herpes virus antigens were found in the sulcular epithelium of approximately 60% of patients with clinically healthy gingiva. In addition, specific antigens for herpes simplex virus (HSV) were found in the sulcular epithelial cells of patients undergoing periodontal treatment. Specific antibodies were also detected in 70-80% of the gingival fluids collected. On the basis of these data we hypothesized that the oral cavity may act as a preferential site for latent HSV. Thus, stressful events such as traumatic dental treatment and tissue damage may elicit herpetic episodes, risking dental personnel. Measures of precaution are indicated for routine dental treatment. PMID- 2562338 TI - Combined hepatocellular-cholangiocarcinoma associated with dermatomyositis. AB - A 56 year old female developed combined hepatocellular cholangiocarcinoma associated with dermatomyositis. Serum tumour markers except for carbohydrate antigen (CA 19-9; 6400 ng/ml) were within normal range. Despite extensive chemotherapy, no clinical response was obtained and the patient's course deteriorated after 4 months. Macroscopically, the liver was mainly occupied by hepatocellular carcinoma but cholangiocarcinoma was found in the hilum. This is the first case of a rare association of combined hepatocellular cholangiocarcinoma and dermatomyositis. PMID- 2562339 TI - Diarrhoea in AIDS patients. PMID- 2562341 TI - [Medical aspects of dietetic fiber]. PMID- 2562340 TI - Changes in urinary electrolytes excretion after injections of ionic solutions into lateral hypothalamic area of rats. AB - Rats submitted to a chronically implanted cannula into lateral hypothalamic area (LHA) were utilized to study the effect of ionic stimulation of KCl, LiCl and CaCl2, sodium free solutions, on renal water and electrolyte excretion. In a general way decreases in natriuresis and kaliuresis following the injections of solutions into LHA were observed. Furthermore, significant increases in diuresis were noted after the injections of the 0.30 M solutions. PMID- 2562342 TI - [Absence of seroprevalence against cytomegalovirus and herpes simplex in a group of patients with coronary atherosclerosis]. PMID- 2562343 TI - [Cardiovascular parasympathetic and orthosympathetic lesions in alcoholism and their association with peripheral polyneuropathy]. AB - Our aim was to identify the damaged baroreceptor reflex in alcoholic patients, and its association to peripheric neuropathy, and the electrophysiological data of primary axon damage and secondary myelinic damage. We measured the R-R interval and the blood pressure resting, after valsalva maneuver, six deep breathing/min and decubitus-biped, changing position, in 22 alcoholic patients and 22 healthy persons. Electromyography was performed, measuring the nerve conduction velocity and peripheral latency time of sciatic-popliteal external and internal nerves. We observed important damage of the cardiac parasympathetic and vascular sympathetic system, probably extended to the sympathetic cardiac system because the median cardiac rate of the alcoholic and the healthy person were similar. These lesions were associated with each other and with the motor neuropathy, with simultaneous development and common pathogenic mechanism. Finally, the R-R response had minimal amplitude during deep breathing without velocity alteration, suggesting an axonal alteration more evident than myelinic damage. PMID- 2562344 TI - [Sublingual enalapril in hypertensive crisis. A preliminary study]. AB - The effects of a new angiotensin I-converting enzyme inhibitor (ACEI), enalapril via sublingual was studied in 16 patients (median age 60.5 +/- 8.7 years). 10 were female and 6 male with hypertension crisis defined as systolic pressure (SP) greater than or equal to 220 mmHg and/or diastolic pressure greater than or equal to 120 mmHg (DP). We observed a significant decrease of SP and DP 20 min after the enalapril administration, without affecting the cardiac rate (p less than 0.001). This decrease was maintained during 30 and 60 min (p less than 0.001). Secondary effects were not observed. We concluded that sublingual enalapril can be a useful drug in hypertension crisis when not an urgent life compromising situation in elderly patients, because of its gradual hypotensive and maintained effect, allowing the commencement of an oral treatment. PMID- 2562345 TI - Antenatal infections and screening. PMID- 2562346 TI - Sexually transmitted pelvic infections and vaginitis. PMID- 2562347 TI - Neoplasms. PMID- 2562349 TI - [Application of isocyanate separating agent for acrylic resin]. AB - The most widely used agents for separating acrylic resin from gypsum molds are the water-soluble alginates, which produce a very thin water- and organic solvent insoluble calcium alginate film on the gypsum surface. In the present study, a new type of separating agent, an isocyanate type coating agent, was developed and it's properties were investigated. This coating agent is composed mainly of trimethylol-propane-tolylenediisocyanate and ethyl-acetate, with a small amount of bis-tributyl-tin oxide as catalyst. At first, a thin film was formed on the surface by application of the agent, and then alkyl-phosphate was applied to this film to ensure easy separation of the cured resins from the molds. The molds treated with this separating agent had a glossy, hard surface. The surface of the cured resins treated in this way was glossier and smoother than that of resin treated with the water-soluble alginates. PMID- 2562348 TI - Resected case of hepatocellular carcinoma associated with lupoid hepatitis. AB - A 65 year old woman with lupoid hepatitis developed hepatocellular carcinoma which was diagnosed at an early stage. She had no history of blood transfusion and serum hepatitis B virus markers were negative. Prednisolone and 6 mercaptopurine were administered for the treatment of lupoid hepatitis. A hepatocellular carcinoma was detected by the elevation of serum alpha-fetoprotein and imaging studies. A tumour, 1.4 cm in diameter, was located in the lateral segment of the left hepatic lobe. It was resected by hepatic subsegmentectomy. Histological study showed a hepatocellular carcinoma of Edmondson type II against a background of posthepatitic cirrhosis. The patient was in good condition 2.5 years after the operation. PMID- 2562350 TI - Tumour markers of hepatocellular carcinoma. PMID- 2562351 TI - Diagnosis of small hepatocellular carcinoma by computed tomography: correlation of CT findings and histopathology. AB - The diagnostic value of computed tomography (CT) scans in small hepatocellular carcinoma (HCC) (less than 5 cm) was studied in 82 patients. Dynamic scan was also made in 66 of them. Combined unenhanced and enhanced scans detected 87% of the lesions greater than 2 cm, but the detection rate was only 25% for lesions less than 1.5 cm. Diagnostic failure was due to isodensity of the mass and to technical artefacts. Diagnosis of the surrounding capsule and internal septa (partition) and demonstration of the typical pattern of density enhancement by dynamic scan proved useful in differentiating HCC from secondary cancers. On unenhanced CT, the density of the interior was subject to the histological changes of tumour such as bleeding, necrosis and fatty metamorphosis. Similarly, enhanced CT showed density changes suggestive of these histological changes. Dynamic scan proved particularly useful for lesions less than 3 cm because the typical density enhancement was frequently demonstrated in the arterial phase. It was concluded that unenhanced CT combined with dynamic scan has a high diagnostic value in small HCC and reflects histological changes. PMID- 2562352 TI - Clinicopathological approach to human liver allograft dysfunction. PMID- 2562353 TI - Hepatocellular carcinoma in cirrhotic liver in Trieste. AB - One hundred consecutive cases of hepatocellular carcinoma (HCC) in cirrhosis observed at autopsy were studied and their pathological aspects were compared with those reported in the literature. The results, which are representative of HCC epidemiology in a geographical area where cirrhosis is mostly due to alcohol abuse, show that similarities in the architectural pattern of HCC and weight of the liver exist between our material and samples with different aetiology and epidemiology. A relationship between the histological grade of HCC and its propensity to metastasize was demonstrated. The reported better prognosis of clear cells per se could not be confirmed, although clear cell HCC occurred exclusively in grade 2. It was also demonstrated that the relationship between grading and staging was strongly influenced by the association of HCC with cirrhosis, which is a fact that is usually overlooked by the common staging (and grading) methods. PMID- 2562354 TI - Correction of metabolic defect by gene transfer into isolated hepatocytes. PMID- 2562355 TI - Polyradiculopathy and sensory ganglionitis due to cytomegalovirus in acquired immune deficiency syndrome (AIDS). PMID- 2562356 TI - Specific immunity to HIV and other retroviral infections. PMID- 2562357 TI - Diagnosis of cytomegalovirus infection of the lung in the acquired immune deficiency syndrome (AIDS): by in situ DNA hybridization. PMID- 2562358 TI - NGF in CNS: experimental data and clinical implications. AB - The presence of beta-nerve growth factor (NGF) and its cell surface receptor (NGF R) in the brain has been well established by a variety of experimental techniques in recent years. In particular, the molecular cloning of NGF and NGF-R as well as the development of sensitive two-site ELISA techniques for determining the levels of NGF and antibodies to NGF-R suitable for immunohistochemistry have led to rapid accumulation of data in this field from many laboratories. A main finding is the function of NGF in the cholinergic neurons of the basal forebrain, expressing NGF receptors and responding to the factor by increased activity of choline acetyltransferase, and the production of NGF in cortical areas and hippocampus comprising terminal areas for the cholinergic projections from the basal forebrain. In addition, findings suggest that additional neurons in the brain and spinal cord may utilize NGF, notably during development and possibly also after lesion of the adult CNS. Moreover, observations indicate that endogenous levels of NGF are lowered in the aged rat brain concomitant with losses of NGF-dependent neurons in the basal forebrain. The involvement of NGF in human neurodegenerative diseases is not established but the application of NGF to degenerating cholinergic neurons in Alzheimer patients may prove useful. A promising approach to achieve this goal is the production of biologically active, recombinant NGF. PMID- 2562359 TI - Bradykinin receptors: characterization, distribution and mechanisms of signal transduction. AB - Bradykinin is a peptide consisting of nine amino acids. It is a member of the kinin family, a class of molecules sometimes considered to be locally acting hormones. Bradykinin acts through cell surface receptors to elicit a series of biological responses, many of which have been well characterized at the whole organ or body level. However, little is known about the bradykinin receptor itself or its mechanisms of signal transduction, its function and its tissue distribution. Increasing evidence suggests that bradykinin is a member of a group of locally produced peptides which may act in a paracrine fashion as microenvironmental modulators of cell proliferation. Evidence for this derives from studies of the interaction between bradykinin and its receptor, receptor effector coupling systems and in vitro studies of the biological effects of bradykinin. These areas, together with questions concerning the nature and number of different types of bradykinin receptors, form the main bulk of current interest in bradykinin research and are the subject of this review. The ability of bradykinin to synergize with other growth regulating ligands will also be considered. PMID- 2562360 TI - Structural and functional aspects of the receptors for platelet-derived growth factor. AB - Platelet-derived growth factor (PDGF) is a 30 kDa dimer of disulfide-bonded A and B chains. Three isoforms of PDGF have been isolated (PDGF-AA, PDGF-AB and PDGF BB). These bind with different affinities and specificities to two structurally related cell surface receptors, viz. the alpha-receptor and the beta-receptor. The receptors are transmembrane proteins with an intracellular, ligand stimulatable protein tyrosine kinase domain. Activation of the receptors is intimately associated with receptor dimerization, and available data suggest that PDGF is a divalent ligand such that one molecule of PDGF binds and dimerizes two receptor molecules. Stimulation of PDGF receptors leads to a cascade of cellular events, which have been shown to require an intact receptor tyrosine kinase activity. However, ligand-induced internalization and degradation of the beta receptor occur essentially independent of the receptor kinase activity. Receptor activation leads to the phosphorylation on tyrosine residues of three enzymes, probably by direct phosphorylation: phospholipase C-gamma, phosphatidylinositol 3' kinase and Raf-1. In certain cells, PDGF beta-receptor expression is inducible such that cells in normal tissue in vivo do not express receptors; only in inflammatory lesions or when cells are explanted in vitro, are receptors being expressed. Transformation by the v-sis oncogene is mediated by an autocrine PDGF like growth factor. Although both the alpha- and beta-receptors are structurally related to the v-fms and v-kit oncogenes, it is not known if the PDGF receptors have a transforming potential. In conclusion, the finding of three isoforms of PDGF that interact with two structurally related receptors implies a finely tuned regulatory network, the role of which in cell growth and transformation remains to be clarified. PMID- 2562361 TI - Beta-adrenergic receptors and catecholamines in acute myocardial infarction. AB - Lymphocyte beta-adrenergic receptor density and plasma catecholamine concentrations were determined in 28 patients with acute myocardial infarction and compared with those in patients with angina pectoris and healthy persons. In patients with acute myocardial infarction beta-adrenergic receptor density was significantly lower (p less than 0.001) and plasma catecholamine levels significantly higher (p less than 0.001) as compared with corresponding values in patients with angina pectoris or healthy persons. beta-adrenergic receptor density in patients with angina pectoris were not significantly different from those in controls. A significant negative correlation between beta-adrenergic receptor density and plasma norepinephrine levels was observed in patients with acute myocardial infarction (r = -0.593; p less than 0.001; r = -0.615; p less than 0.001 respectively). It is suggested that decreased beta-adrenergic receptor density is a consequence of elevated plasma catecholamine levels in patients with acute myocardial infarction. It has been well documented that acute myocardial infarction is associated with enhanced activity of the sympathetic nervous system. Several studies have already been done showing that urinary excretion of catecholamines and plasma catecholamine concentrations are raised in the acute phase of myocardial infarction. Particularly high levels of plasma catecholamines appeared to be related to the severity of clinical course of myocardial infarction and were found in patients with cardiogenic shock, heart failure and arrhythmias. It is of interest that the peak elevation of plasma catecholamines correlated with the extent of myocardial damage as reflected by peak plasma CK activity and also correlated with acute and long-term mortality.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562362 TI - [Blood antigens and specific anti-core and anti-envelope antibodies as markers of the course of HIV infection]. AB - The antigenemia and the patterns of antibodies to core protein (p24) and envelope glycoproteins (gp41, gp120) have been investigated in 81 patients with Human Immunodeficiency Virus (HIV) infection followed prospectively for 24 months. HIV antigen was detectable in 23 (28.4%) patients at entry to the study (13/13 with AIDS and 10/23 with ARC) and in 33 (40.7%) at the end (25/28 with AIDS, 5/12 with ARC e 3/14 with LAS). Anti-p24 were positive in 51 (63.0%) patients at the entry (26/30 symptomless, 13/15 with LAS e 12/23 with ARC) and in 41 (50.6%) at the end of the study (23/27 symptomless, 9/14 with LAS, 7/12 with ARC e 2/28 with AIDS). All patients were positive for anti-gp41 and showed no significant changes in the antibody titers during the two years of follow-up; by contrast, anti-gp120 was undetectable in most patients (26/28) with AIDS. Clinical progression in a high proportion of patients was associated with the appearance of HIV antigen, with the decline of anti-p24 titers and with no antibody reactivity to gp120 glycoprotein. PMID- 2562363 TI - Essential mixed cryoglobulinemia: a report on 14 cases. AB - Fourteen cases of Essential Mixed Cryoglobulinemia (EMC) are described in this report. Clinical and laboratory manifestations in our patients were similar to those previously reported in literature, although involvement of the peripheral nervous system was much more prevalent in our series. We suggest that peripheral neuropathy should be systematically searched in EMC patients. PMID- 2562364 TI - Cloning and expression of delta-endotoxin gene of Bacillus thuringiensis in Escherichia coli. AB - Fragments (larger than 4 kb) of partially Sau 3AI-digested plasmid DNA from Bacillus thuringiensis subsp. kenyae 7404 and kurstaki HD-1 were cloned into the BamH I site of pBR322. Four transformants, containing the corresponding delta endotoxin gene and producing proteins that reacted with antiserum against crystalline protein, were selected on the basis of results from in situ colony hybridization, radioimmunologic screening, and Western blot analysis. Three of those tested, i.e., the lysates of one transformant (TK89) carrying the delta endotoxin gene of B.t. 7404 and two transformants (TH12 and TH48) carrying the delta-endotoxin gene of HD-1, were toxic for tobacco budworm (Heliothis assulta) and armyworm (Leucania separata Walker) caterpillars. This is the first report on cloning of the delta-endotoxin gene from B.t. subsp. kenyae, which is different in serotype from the well-studied subsp. kurstaki. PMID- 2562366 TI - Varying protocols for malignant brain tumors. PMID- 2562365 TI - Biological basis of omeprazole therapy. AB - There are two means of reducing acid secretion. The best studied is inhibition of stimulation of the parietal cell. There are three major types of receptors that activate secretion by this cell and two classes of receptor antagonists, as well as at least two intracellular messenger pathways. The receptors are for histamine (H2 subtype), acetyl choline (M2 subtype) and gastrin. Antagonists of these receptors include the H2-antagonist class (Tagamet, Zantac and Pepcid), the M1 muscarinic antagonists (pirenzepine, telenzepine) and the gastrin antagonist, proglumide. The major pathway for stimulation appears to be the H2-receptor, since this is the only receptor that stimulates adenylate cyclase, and both acetyl choline and gastrin release histamine locally within the gastric mucosa. However, these agonists elevate intracellular calcium, which has a partially independent action on acid secretion. Accordingly, the most efficacious type of receptor antagonist will be of the H2 class, which is borne out by clinical experience. Prostaglandins of the E type prevent adenylate cyclase stimulation by histamine and are also effective antisecretory agents. It will be difficult to abolish acid secretion entirely by a single receptor antagonist, although longer acting H2-antagonists should show clinical superiority to short-acting antagonists of this type. An alternative approach to acid suppression is to block the terminal step of acid secretion, the gastric proton pump (H+, K(+)-ATPase). This enzyme is virtually unique to the parietal cell and, when active, forms a very acidic space within the parietal cell called the secretory canaliculus. Activation of acid secretion involves several steps. The enzyme is present in cytosolic membranes when the cell is at rest and moves to the membrane of the secretory canaliculus when stimulated. Simultaneously, there is an increased permeability of potassium chloride (KCl), which allows presentation of K+ to the luminal surface of the pump and H+ for K+ exchange. The result is the secretion of HCl into the canaliculus, and hence into the gland lumen and then the stomach. There are two classes of pump inhibitors. One class is K+ competitive and relatively selective for the H+, K(+)-ATPase, as exemplified by SCH28080. This class has not yet been used in man. The other class is specific to the functioning H+, K(+)-ATPase in the stomach. It is exemplified by omeprazole (Losec). This compound is a weak base with a pKa of 4. In the unprotonated, uncharged form it will penetrate cell membranes and, at pH less than 4, it becomes protonated and therefore charged.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2562367 TI - [The risk of retrovirus transmission by blood transfusions]. PMID- 2562368 TI - [Zinc in patients with ascites]. AB - The concentrations of zinc (Zn) in blood and ascitic fluid (AF) in 48 patients with ascites (alcoholic liver cirrhosis: 32; liver carcinoma associated to cirrhosis: 7; different malignant neoplasias: 9). In addition, total protein, albumin and alpha 2-globulin as well as zinc, were determined in ascitic fluid. There was a decrease of zinc levels in serum, total protein, albumin and alpha 2 globulin in ascitic fluid of cirrhotic patients compared to neoplastic patients. The zinc concentration in ascitic fluid was related to the concentration of total protein, albumin, alpha 2-globulin. There is a parallel correlation between seric zinc and AF zinc. PMID- 2562369 TI - [The action of naloxone on monocyte chemotaxis in heroin addicts]. AB - Intravenous (I.V.) drug users possess a decrease of the monocyte chemotaxis directly related to drug usage. We study the effects of naloxone on monocyte chemotaxis in I.V. drug users, evaluating the chemotactic index and the cellular level of cAMP. The incubation of monocytes with naloxone did not modify the chemotactic index nor the cellular levels of cAMP, the first remaining low and the second high, this being statistically significant compared with the control group. We comment on the possible causes of naloxone's lack of effect and we suggest the role of cAMP in causing the chemotactic defect in this group of patients. PMID- 2562370 TI - [Thymidine kinase: a serum marker in clinical medicine]. AB - The need for new seric markers of tumoral or viral activity to enable an early diagnosis or staging has brought about a proliferation of research in new markers. The deoxi-thymidine-kinase (DTK) is an enzyme of DNA synthesis during cellular proliferation. We review the literature, concluding that DTK is a good seric marker in some neoplasias and some viral diseases because of its specificity, low cost and rapid testing in multiple samples. PMID- 2562371 TI - Interferon-gamma down-regulates membrane adenylate cyclase activity of a murine macrophage-like cell line (P388D1). AB - Effects of recombinant murine interferon-gamma (rIFN-gamma) on the membrane adenylate cyclase of a murine macrophage cell line (P388D1) were investigated in order to explore the nature of a signal transmitted by IFN-gamma receptor. Following the incubation of P388D1 cells with 40 U/ml of rIFN-gamma, the intracellular level of cAMP gradually increased about twofold over the control level within 60 min, and then began to gradually decline to about half the control level by 24 h incubation. The initial rise in cAMP level appeared to be due to the modest activation of adenylate cyclase and not due to the inhibition of cAMP-phosphodiesterase. Later decrease of intracellular cAMP may be due to quantitative down-regulation of the adenylate cyclase system. The basal enzymatic activity of the membrane prepared from P388D1 cells exposed to IFN-gamma for 24 h was found to be reduced to about 20% of that of the control membrane. However, the quality of the adenylate cyclase system appeared unchanged, because the relative degree of the response of the down-regulated membrane adenylate cyclase to prostaglandin PGE2, NaF, guanylimidodiphosphate (GppNHp), choleara toxin (CT), or forskolin was found to remain unchanged. This quantitative down-regulation of adenylate cyclase must be due to the action of rIFN-gamma, since the prior treatment of rIFN-gamma with either acid (pH 2) or monoclonal anti-IFN-gamma antibody inhibited the ability of IFN-gamma to induce the down-regulation. The rIFN-gamma-induced down-regulation is a reversible process, since the adenylate cyclase activity of the membrane was found to be restored when the rIFN-gamma exposed cells were cultured for 72 h in the absence of rIFN-gamma. In addition, the 48 h-incubation of P388D1 cells with rIFN-beta or IFN-alpha was found not to significantly affect the membrane adenylate cyclase system. PMID- 2562372 TI - [Effect of xanthine oxidase inhibitors on the prognosis of acute intestinal ischemia]. AB - During the last few years, the scientific field has focused its attention on the pathogenic role of free radicals in the process of ischemia-revascularization. It is a well-known fact that xanthine oxidase is an important source of tissular free radicals. Bearing this in mind, we designed an experimental protocol to analyse the effect of allopurinol (a xanthine oxidase inhibitor) in the survival of rats after the occlusion of the superior mesenteric artery during a period of 90 minutes and its action on the superoxide radical liberation. The concentration of oxipurinol and allopurinol in the ischemic area (intestine), liver and blood were measured. We concluded that the administration of allopurinol increased the survival rate, which is correlated to higher concentrations of allopurinol and oxipurinol in the inner part of the intestinal cells. A correlation between the survival rate and superoxide radicals was not found. PMID- 2562374 TI - [Opsoclonic cerebellar disease. A paraneoplastic syndrome sensitive to chlormethiazole]. AB - Opsoclonus is a very rare ocular dyskinesia associated to a viral infection or neoplasms, being neuroblastoma in children or carcinoma in adults. A case of opsoclonus as a paraneoplastic symptom preceding the diagnosis of breast carcinoma is presented. The abnormal movements had a complete response to the administration of chlormethiazole. PMID- 2562373 TI - [Adult respiratory distress produced by Pneumocystis carinii in a patient without AIDS]. AB - A case of a patient receiving chemotherapy because of breast cancer who developed adult respiratory distress caused by Pneumocystis carinii pneumonia is presented. The evolution was good after treatment with 20 mg/kg/day of trimethoprim and 100 mg/kg/day of sulfamethoxazole. PMID- 2562375 TI - [Small-cell lung carcinoma and pancreatitis]. PMID- 2562376 TI - [Enalapril-induced cough]. PMID- 2562377 TI - Diminished expression of the T cell receptor on the expanded lymphocyte population in MRL/Mp-lpr/lpr mice. AB - MRL mice homozygous for the recessive lpr gene develop an accelerated autoimmune syndrome and massive lymphadenopathy. Because the function of the expanded lymph node population is unclear, we have studied the subunits of the T cell receptor for antigen (TcR). DNA and RNA were prepared from MRL/Mp-lpr/lpr (lpr) and congenic MRL/Mp(-)+/+ (+/+) mice by standard techniques and studied by Southern blot, northern blot, and dot blot analysis using the cDNAs TT11, specific for the TcR alpha chain; 86T5, specific for the TcR beta chain; and T3 delta; specific for the subunit of the T3 molecule. Surface protein was immunoprecipitated with antisera 8177, which recognizes TcR framework determinants, and resolved by diagonal SDS-PAGE. FACS analysis was performed with a monoclonal antibody to murine T3, and with the KJ16-133 and F23.1 monoclonal antibodies, which recognize determinants encoded by the V beta 8 subfamily of beta chain variable region genes. When compared with +/+ controls, surface TcR density as detected by immunofluorescence using all three antibodies was significantly diminished on lpr spleen and lymph node cells, as well as on lpr lymph node cells which had been depleted of L3T4+ and Ly2+ cells by negative selection. There appeared, however, to be selective expression of the genes encoding the epitopes binding F23.1. Southern blot analysis of DNA showed polyclonal rearrangements of the TcR beta chain genes. There were increased alpha, beta, and T3 delta RNA transcripts in the double negative lymph node cells. The paradoxical decrease in TcR surface expression in the setting of large quantities of full length transcript is yet to be explained. PMID- 2562378 TI - Beta-adrenergic cardiac antibody in autoimmune myocarditis. AB - Balb/c mice were immunized with homologous heart in complete Freund's adjuvant to induce autoimmune myocarditis. The myocarditis was characterized by lymphomononuclear infiltration, electrocardiographic abnormalities and antimuscle antibodies by indirect immunofluorescence. In this paper, we demonstrate that the IgG present in autoimmune myocarditis mice is able to bind to beta adrenoreceptors of the heart and also induce a biological effect inhibiting the contractile action of exogenous norepinephrine. Auto-immune IgG inhibited the binding of (3H)-dyhidroalprenolol to a beta-adrenergic receptor of purified myocardial membranes behaving as non-competitive inhibitor. This IgG also exerted a non-competitive inhibition upon the mechanical effect of exogenous norepinephrine. The recognition appears to be organ specific, because the autoimmune myocarditis IgG did not bind to beta-lymphocyte, lung and fat adrenoreceptors. The autoimmune IgG inhibited the stimulatory action of isoproterenol on cAMP levels, behaving as a beta-adrenergic antagonist. PMID- 2562379 TI - The "in vitro" effects of lithium on the immune system. AB - This study has examined the "in vitro" effects of Lithium Carbonate on the immune system at low (10(-3), 10(-2)mM) and therapeutic (0.5-1.5 mM) concentrations. Lithium, in the presence of a range of mitogens, was found to increase the incorporation of 3H-thymidine into peripheral blood mononuclear (PBM) cells. At concentrations greater than 1 mM IL2 production was also enhanced. Lithium was also found to increase IgG and IgM production--an estimate of B cell function, the effects being greatest at concentrations within the therapeutic range. However at these levels Lithium inhibited cAMP production. Whether Lithium acts individually on these processes or whether one reaction is merely the result of another is unclear at present. PMID- 2562380 TI - Long-term response in a patient with ITP following low dose anti-D immunoglobulin therapy. AB - A patient with chronic ITP relapsed after conventional therapy but following an infusion with low dose anti D (Rho) immunoglobulin, she entered a remission which has now lasted 10 months. This is difficult to explain on the basis of long term macrophage Fc receptor blockade and suggests an alternative mechanism of action. PMID- 2562381 TI - A human-mouse hybridoma which secretes monoclonal thyroglobulin autoantibody with properties similar to those of the donor patient's serum autoantibody. AB - Human monoclonal antibodies produced by Epstein Barr (EB) virus transformation and/or cell fusion are frequently IgM antibodies which tend to cross react with a range of antigens and often bear little relationship to the highly specific IgG antibodies associated with human autoimmune disease. By fusing EB virus transformed B lymphocytes from a Hashimoto patient with a mouse myeloma line and selecting for synthesis of IgG class thyroglobulin (Tg) antibody, we have developed a hybridoma (VB/5) secreting Tg antibody of IgG2 subclass and lambda light chain type which has the characteristics of a monoclonal antibody on isoelectric focussing. The antibody has a high affinity for human Tg and recognises Tg from other primates but not non-primate Tg. However, it does not react with human thyroid peroxidase or a panel of other autoantigens. In terms of affinity constant, functional affinity and affinity heterogeneity, the antibody closely resembles the IgG2 lambda Tg antibody present in the serum of the Hashimoto patient whose B lymphocytes were used to develop the hybridoma. In addition to providing a useful reference standard for Tg antibody IgG subclass assays, VB/5 antibody and the hybridoma line provide a valuable starting point for detailed studies of Tg autoantibodies and the genes coding for the variable regions of their heavy and light chains. PMID- 2562382 TI - The En/Spm transposable element of Zea mays. PMID- 2562383 TI - Physical characterization of mitochondrial DNA from cotton. PMID- 2562384 TI - Cytochrome oxidase subunit III gene from soybean mitochondria. PMID- 2562385 TI - Therapeutic potential of fish oil in the treatment of ulcerative colitis. AB - In a pilot study six patients with active ulcerative colitis and six healthy controls were given fish oil (MaxEPA) containing 3-4 g of eicosapentaenoic acid daily for a period of 12 weeks. There was a significant improvement in the patients' symptoms and histological appearance of the rectal mucosa by the end of the treatment period. There was significant fall in neutrophil chemiluminescence during treatment in patients, whereas no change was observed in the control group. Neutrophil leukotriene B4 levels fell significantly during treatment. Serum from patients receiving fish oil was significantly less chemotactic for neutrophils compared with control serum. Eicosapentaenoic acid inhibited neutrophil chemotaxis and chemiluminescence in vitro. The omega-3 fatty acids, which occur naturally in fish oils, may exert a beneficial effect by decreasing the production of inflammatory mediators. PMID- 2562386 TI - [The air space: alveolar pattern]. PMID- 2562387 TI - [Use of guar gum as a supplement to the usual diet in type 2 diabetes. A long term study]. AB - The effects of diet supplementation with guar rubber in granules (15 g/day, divided in three doses with the major meals) were compared during 3 months with a control group in a randomized study. Thirty-five type II diabetic patients were randomized into two groups (guar and control) of 16 and 19 patients, respectively. The patients in the guar group showed an improvement in the carbohydrate and lipid metabolism as compared with the control group; however, in our opinion, this improvement was not important enough to recommend the routine use of guar rubber in the diet of the diabetic patient owing to its poor acceptance. PMID- 2562388 TI - Autoantibodies to neurons and to the cytoskeleton in small cell carcinoma with paraneoplastic sensory neuropathy. AB - Autoantibodies to neurons and to the cytoskeleton were demonstrated in the serum of a patient with small cell carcinoma of the lung (SCCL) and paraneoplastic sensory neuropathy. The serum reacted by immunofluorescence with the nuclei of neurons, and with cytochalasin B-sensitive "stress fibres" of cultured cells. The serum also reacted by immunofluorescence with the nuclei of some cultured cell lines. Immunoblotting experiments with brain tissue, with SCCL, HeLa and other cultured cells showed that the serum reacted with 1-4 bands of 35-39 kDa apparent m.w. Two dimensional immunoblotting showed that these molecules had a neutral pI. Antibodies, affinity-purified by elution from the 35-39 kDa bands, gave staining of the nuclei of neurons and of cultured cells and immunoblotted the same 35-39 kDa antigens. These observations show that the anti-neuronal autoantibody reacts not only with neurons and with SCCL but also with some cultured cell lines. Molecular mimicry has been invoked as the basis for the reactivity of this autoantibody with exposed epitopes on SCCL and on neurons. PMID- 2562391 TI - Plasma amino acid patterns in portal hypertensive patients. PMID- 2562390 TI - Monoclonal autoantibodies derived from multiple sclerosis patients and control persons and their reactivities with antigens of the central nervous system. AB - Peripheral blood B lymphocytes of multiple sclerosis (MS) patients and control persons were transformed with Epstein-Barr virus. Antibody production of transformed cells against isolated human myelin was investigated by enzyme-linked immunosorbent assay (ELISA). Cells producing reactive antibodies were cloned and propagated to produce monoclonal antibodies (mAbs). These mAbs did also react with acetone fixed frozen sections of normal human white matter, as determined by indirect immunofluorescence staining. Some of the mAbs derived from MS patients and a control person with a central nervous system cyst agglutinated liposomes made from lipids of a chloroform/methanol extract of human myelin, whereas mAbs derived from four glioma patients were negative in these tests. The reactive antibodies were investigated further using agglutination tests with liposomes made from pure auxiliary lipids (cholesterol and lecithin) or containing in addition either galactocerebroside, sulfatide or a mixture of bovine brain gangliosides. The great majority of myelin liposome agglutinating antibodies reacted with all types of liposomes, including those made from pure auxiliary lipids. Investigations by ELISA suggest that phospholipids are the reactive components, at least for some of these mAbs. Some antibodies reacted with liposomes containing galactocerebroside or sulfatide, others only with sulfatide containing liposomes. Antibodies showing these specificities were only obtained from MS patients. PMID- 2562389 TI - T cell recognition of epithelial self. AB - The presentation of self-antigens to circulating T cells is a critical, precipitating event in the induction of autoimmune injury in parenchymal organs. Epithelia expressing these self-antigens are thought to release such moieties for reprocessing by traditional antigen-presenting cells within the lymphoid system. We now demonstrate, however, that some epithelium possess novel functional mechanisms for presenting their own antigens to a responsive, syngeneic T cell repertoire. The presentation of these self-antigens occurs in the context of MHC class II molecules and depends on CD4 associative-recognition determinants. Our findings strongly suggest that organ epithelium may directly activate cell mediated events to produce autoimmunity through self-recognition. PMID- 2562392 TI - Vascular plexus around intrahepatic large bile ducts in normal livers and portal hypertension. PMID- 2562393 TI - A possible effect of gastric microcirculation on the different responses of surgical treatments for hepatocellular carcinoma with oesophageal varices. PMID- 2562395 TI - Additional nodulation genes on the Sym plasmid of Rhizobium leguminosarum biovar viciae. AB - A Rhizobium leguminosarum biovar viciae strain lacking a 40 kb DNA region of the Sym plasmid pRL1IJ to the left (3' side) of gene nodE failed to nodulate Vicia sativa plants. Therefore this DNA region was investigated for the presence of additional nodulation genes. Complementation experiments indicated that the DNA region to the left (3' side) of nodE is functionally homologous between R. leguminosarum bv. viciae and R. leguminosarum bv. trifolii. In this DNA region, three nodulation genes were identified, nodT, nodM and nodL. TnphoA insertions in the nodT gene, about 4.5 kb to the left of nodE, lead to a delay in nodulation on Trifolium subterraneum, but not on V. sativa plants. TnphoA insertions in gene nodM have no detectable influence on nodulation. Finally, TnphoA insertions in the nodL gene affected nodulation so that only rarely nodules were induced on the inoculated plants. The nucleotide sequence of this gene is presented. On the basis of the sequence a membrane integrated protein is predicted with a molecular weight of 20.1 kDa. Microscopical analysis of the infection process by nodL mutants indicate a role for nodL in maintaining the stability of the infection thread. Experiments using transcriptional lacZ fusions suggest that nodL belongs to the same transcriptional unit as nodF,E. Very low expression of nodL seems to be sufficient for biological activity. PMID- 2562394 TI - Stable co-transformation of maize protoplasts with gusA and neo genes. AB - An efficient co-transformation protocol using polyethylene glycol was developed for Zea mays L. (cv. A188 x BMS) protoplasts isolated from suspension culture cells. Co-transformation was accomplished by using plasmid constructions containing beta-glucuronidase (gusA) or neomycin phosphotransferase (neo) gene coding sequences; both were under control of the CaMV 35S promoter. Protoplast culture and transformation conditions were optimized to assure efficient recovery of transformed cells. The overall efficiency of transformation was 1 x 10(-4) (calculated per viable protoplast plated). Among kanamycin-resistant lines, 50% showed a high level of GUS activity (above one unit). Southern blot hybridization confirmed the presence of numerous gusA and neo coding sequences in the maize genome. In two analyzed lines, integrated sequences appeared to be organized in tandem head-to-tail repeats. Results also indicated that the integrated sequences were partially methylated. PMID- 2562397 TI - Ganciclovir for cytomegalovirus infection. PMID- 2562398 TI - Lisinopril--another ACE inhibitor. PMID- 2562396 TI - A comparative study of Tam3 and Ac transposition in transgenic tobacco and petunia plants. AB - Transposition of the Anthirrinum majus Tam3 element and the Zea mays Ac element has been monitored in petunia and tobacco plants. Plant vectors were constructed with the transposable elements cloned into the leader sequence of a marker gene. Agrobacterium tumefaciens-mediated leaf disc transformation was used to introduce the transposable element constructs into plant cells. In transgenic plants, excision of the transposable element restores gene expression and results in a clearly distinguishable phenotype. Based on restored expression of the hygromycin phosphotransferase II (HPTII) gene, we established that Tam3 excises in 30% of the transformed petunia plants and in 60% of the transformed tobacco plants. Ac excises from the HPTII gene with comparable frequencies (30%) in both plant species. When the beta-glucuronidase (GUS) gene was used to detect transposition of Tam3, a significantly lower excision frequency (13%) was found in both plant species. It could be shown that deletion of parts of the transposable elements Tam3 and Ac, removing either one of the terminal inverted repeats (TIR) or part of the presumptive transposase coding region, abolished the excision from the marker genes. This demonstrates that excision of the transposable element Tam3 in heterologous plant species, as documented for the autonomous element Ac, also depends on both properties. Southern blot hybridization shows the expected excision pattern and the reintegration of Tam3 and Ac elements into the genome of tobacco plants. PMID- 2562399 TI - DNA replication in mammalian cells: insights from the SV40 model system. PMID- 2562400 TI - Communicating hydrocephalus occurring in the postoperative course of glioblastoma multiforme. AB - We experienced a case of glioblastoma multiforme which exhibited dementia, gait disturbance, headache, and urinary incontinence six months after subtotal removal of the tumor. These symptoms were not due to tumor recurrence, but to communicating hydrocephalus. Communicating hydrocephalus in cases of malignant brain tumors has not often been reported. We discuss the development of this abnormality. PMID- 2562401 TI - The potential of hormone receptors in the treatment of various cancers. AB - Breast cancer, prostate cancer, endometrial cancer, and lymphocytic leukemias may possess steroid hormone receptors that can predict a high probability of response to the appropriate hormone when the receptor is present. The presence or absence of receptor may also be an important prognostic variable and may aid in the selection of patients for appropriate adjuvant therapy. Although putative receptors have been described in many other tumors, their clinical significance is questionable because these tumors generally do not respond to hormonal therapy. In the future, steroid receptors may enable us to target drugs, radioisotopes, or other molecules to tumors by linking these drugs to steroid hormones. PMID- 2562402 TI - [Transmembrane controls in cultured human thyroid carcinoma]. AB - It is well known that many of thyroid carcinoma are capable of responding to TSH, but our studies shown that there are some alteration in this responsiveness. The adenylate cyclase responsiveness to TSH was usually greater in thyroid carcinoma than in adjacent histologically normal thyroid tissue. The level of increased response of adenylate cyclase were correlated with the level of enhanced expression of ras oncogene product p21 assessed by Western blotting analysis. The TSH induced desensitization of adenylate cyclase was not observed in some differentiated carcinoma. This loss of desensitization may be reflect the change in ADP-ribosylable Gi protein. In the differentiated carcinoma, the capacity of EGF receptor was higher than that in normal thyroid. The EGF binding to cultured carcinoma cells did not increase in response to TSH. These altered properties of transmembrane control in human thyroid carcinoma may be related to the neoplastic growth. PMID- 2562403 TI - [Establishment and characterization of a tumor marker producing cell line (JR-1) derived from a gastric scirrhous cancer]. AB - Despite the importance of in vitro study of gastric cancer, the established cell lines derived from human gastric carcinoma are very few. We have recently established a new cell line derived from human gastric cancer which has the ability to produce several tumor markers. This cell line has been designated JR 1. The cancer cells were obtained from the cerebrospinal fluid of a 37 year-old female patient who had metastatic brain tumor of the poorly differentiated gastric adenocarcinoma. The cells were inoculated into the tissue culture flask containing Ham's F-12 medium supplemented with 10% fetal bovine serum, antibiotics. Within 24 hours, the cells attached to the surface of the flask and started to grow. The first subcultures were performed at 1 week, and subsequent subcultures have been done once a week. This cell line has been maintained for more than 15 months through 60 passages with a stable growth. Chromosome analysis of the cells was performed. The doubling time of the 20th passage was 72 hours. Under phase contrast microscopy, monolayered pavement-like cell arrangement was observed. PAS staining showed intracellular mucin granules. Transmission and scanning electron microscopy revealed spindle-shaped cells with numerous microvilli and fine projections as well as intracellular granules, indicating mucin. Tumor markers produced by this cell line were CEA, CA19-9, TPA and Procollagen III. PMID- 2562404 TI - [Establishment of acinar cells from human salivary gland tissue by transformation of SV40]. AB - A piece of minor salivary gland tissue obtained from the lip of a 16 years old female was minced to about 3 mm3 by fine pincettes and cultured with 10% FCS containing MEM supplemented with EGF (10 ng/ml), fungizon (10 mcg/ml) and kanamycin (60 mcg/ml) in a 5% CO2 incubator. Many small bubbles of saliva were found on the surface of the fragments after 3 to 4 days of incubation and outgrowth of cells from the fragments was observed from 7 days of incubation. Monolayer cells of the outgrowth were trypsinized and passaged with fresh culture medium. At the 8th passage, monolayer cells were infected with SV40 at moi 100PFU/cell. After 18 hour-incubation, the suspension of the infected cells was incubated at densities of 10(4) and 10(3) cells/dish within 0.33% agar containing culture medium. Transformed colonies were picked up from soft agar medium and 3 of the 28 colonies were identified as being acinar cells of the salivary gland, since secretory granules and mucosubstances were specifically proved in the cytoplasm of these cells after 2 to 4 days of incubation. One of the typical clone cells was named HA-16 cells. However, the appearances of the secretory granules and mucins in the cytoplasm of the HA-16 cells depended on the cellular growth cycle, i.e. secretory granules and mucins were not found in the growing cells (G1-S-G2-M phase) but many secretory granules and mucins could be recognized in the non-dividing cells (G0 phase).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562405 TI - [Coupling of GTP-binding protein to inositol phospholipid metabolism in chemoattractant-stimulated guinea pig peritoneal exudate macrophages]. AB - The involvement of GTP-binding protein in inositol phospholipid metabolism in guinea pig peritoneal exudate macrophages stimulated with the chemoattractant N formyl-Methionyl-Leucyl-Phenylalanine (fMLP) was examined. The GTP analog, guanosine 5'-[gamma-thio]triphosphate (GTP gamma S) caused a dose-dependent increase in the formation of inositol triphosphate in membranes of macrophages. This effect was specific for GTP and its analog. fMLP-induced inositol phospholipid turnover was markedly inhibited by the prior exposure of macrophages to 100 ng/ml of pertussis toxin (PT). Likewise, the pretreatment of macrophages with 100 ng/ml of PT evoked the inhibition of the increase in the intracellular free Ca2+ concentration and the spreading of macrophages induced by fMLP. These actions of PT were not associated with an alteration in the cellular concentration of cyclic AMP. Incubation of the membranes of macrophages with [32P]NAD and PT resulted in the ADP-ribosylation of a 41,000 Da protein. This ADP ribosylation was diminished by the prior incubation of the membranes with 100 microM GTP gamma S plus 1 mM MgCl2, indicating that the 41,000 Da protein may be the alpha subunit of a GTP-binding protein. Moreover, there was a parallel between the time course of the ADP-ribosylation of intact macrophages by PT and the inhibition of the increase in intracellular free Ca2+ concentration as well as of the enhancement of the spreading of macrophages. These results suggest that the 41,000 Da protein, a GTP-binding protein, mediates the fMLP-stimulated inositol phospholipid metabolism. PMID- 2562406 TI - [Effects of chewing exercise on the maximum biting force and chewing performance]. AB - Maximum biting force and chewing performance were measured in adult subjects before and after four-weeks training by newly devised "Chewing Ability Enhancing Substances (CAES)". The CAES is made of glucomannan. The number of chewing strokes and chewing time until the last swallowing action are much larger when chewing CAES than those of other usual eating materials. By four weeks training using CAES, the maximum biting force and chewing performance of the subject were clearly increased. However, this increased chewing ability began to return to the control level gradually 2 weeks after the cessation of the training. PMID- 2562407 TI - Effect of stannous ion on the spontaneous transmitter release from motor nerve terminals of the frog. PMID- 2562408 TI - [Molecular-epidemiological and phylogenic analysis of herpes simplex virus type 1 from seven areas in Japan]. AB - Two-hundred and seventy-five epidemiologically unrelated herpes simplex virus type 1 (HVS-1) strains isolated from seven areas (Sapporo, Akita, Nagoya, Kyoto, Tottori, Kagawa and Kurume) in Japan were compared with the standard laboratory strain F, based on cleavage analysis of HSV-1 DNAs with three restriction endonucleases, BamHI, KpnI and SalI. The results obtained were as follows: 1) Using gains and losses of 19 cleavage sites selected from 114 sites, the total of 275 strains could be classified into 87 distinct cleavage patterns. Also, it was found that the isolates were clustered in four predominant patterns, the pattern 27, 6, and 76, containing 62, 24, 15 and 12 strains, respectively. 2) There were highly significant differences in the incidence of isolates classified into the pattern 27 that were obtained in Kagawa as compared with those in Sapporo, Akita, Nagoya and Kurume, and those in Tottori as compared with those in Sapporo, Akita, Nagoya and Kurume. There was also a significant difference in the incidence of isolates classified into the pattern 76 that were obtained in Kagawa as compared with those isolated in Kurume. 3) There were significant correlation coefficients (p less than 0.05) between some stets, e.g. loss of the site between A and A' and gain of a site in A cleaved with BamHI, in every pair of 18 cleavage sites. 4) The phylogenic tree of 87 patterns based on genomic similarities of the Japanese HSV-1 isolates was established, and it was considered that HSV-1 isolates from Japanese could be phylogenetically classified into two to six major groups. These results suggest that HSV-1 strains have mutated and evolved independently by the transmission of the viruses among geographically separated hosts over an extremely long period, and that the genetically characteristic variants have accumulated and persisted in the present Japanese population. PMID- 2562409 TI - [A study on the formation of apatite crystallized with gel method]. AB - About apatite produced with a silicahydro gel method using calcium nitrate (group I) or calcium chloride (group II) and a gelatin gel method by use of calcium nitrate (group III) or calcium chloride (group IV), the formative volume as well as the formative condition of a periodic-layered precipitate (Liesegang ring), the pH measurement, calculation of Ca/P ratio, an estimation of the chlorine ion, morphological observation with a scanning electron microscope, qualitative analyses by X-ray diffraction (identification, crystallite size, lattice imperfections, lattice constants) and the composition analysis by infrared absorption spectroscopy were carried out to elucidate the formation of apatite using the gel method. The result showed that there were no distinct differences between group I-II and group III-IV, and it is suggested that it is possible to form satisfact fluorapatite with a gel method using calcium chloride as well as calcium nitrate. PMID- 2562410 TI - [Serological classification of Bacteroides gingivalis and purification of group specific antigens]. AB - Bacteroides gingivalis were serologically investigated by the immunodiffusion technique using autoclave-extracted antigens and antisera against whole cells. Fifteen strains of B. gingivalis used in this study, were classified in two distinct serological groups. Eleven strains include strain 381, ATCC 33277 were classified Group A and four strains include strain Su63 were classified in Group B. The group specific antigens from B. gingivalis strain 381 (Group A) and Su63 (Group B) were partially purified from autoclave extracts by 75% ethanol precipitation and anion exchange chromatography, and were identified as polysaccharides composed of glucose (82.3%, 23.7%), rhamnose (7.1%, 12.7%), galactose (4.9%, 18.2%), mannose (2.3%, 1.7%), glucosamine (0.4%, 28.0%) and galactosamine (3.0%, 15.3%). PMID- 2562411 TI - Inhibitors of DNA gyrase. PMID- 2562412 TI - [Cushing syndrome caused by macronodular adrenal hyperplasia, independent of ACTH: report of a case]. AB - A 45 year old female with Cushing's syndrome due to non-ACTH dependent bilateral adrenal macronodular hyperplasia (AMH) is reported. The diagnosis of Cushing's syndrome was based on the typical clinical features and on the demonstration of high urinary levels of free cortisol (microF) (630 micrograms/24 h) and 17 hydroxysteroids (17-OHS) which failed to suppress during the Liddle test (17-OHS, (mg/g creat), Basal: 68.5, post Dexamethasone 2 mg: 59.6 and post Dexamethasone 8 mg: 69.9). The adrenal CT scan showed bilateral multinodular enlargement while the pituitary CT scan was normal. Due to the presence of severe hypertension (240/150) and depression, the patient was treated with ketoconazole (800 mg/d) during 8 months achieving eucortisolism (microF 14-39 micrograms/24 h); however, plasma ACTH was not detectable at the end of this period. A bilateral adrenalectomy was performed, and both adrenals showed multiple nodules (0.3-4.5 cm in diameter) and weighed 136 and 31 g respectively. The lack of suppression of the 17-OHS with 8 mg of Dexamethasone, the persistence of an adequate inhibition of cortisol biosynthesis with ketoconazole, and the absence of plasma ACTH suggest that the patient had a non-ACTH dependent AMH. The possible pathogenic factors involved in this case are discussed. PMID- 2562413 TI - [Enalapril in essential arterial hypertension]. AB - The hypotensive effect of enalapril, a converting enzyme inhibitor, given as a single agent once daily, was evaluated prospectively over a 16 week period in 20 subjects. One patient abandoned therapy and 2 were withdrawn (increased creatinine levels in one and the need for associated therapy in the other). Side effects developed in 3 patients: creatinine elevation, skin rush and a "salty taste", respectively. A significant decrease in blood pressure (p less than 0.01) attaining normal levels in 6 out of 17 patients was observed. Normal blood pressure levels were not reached in severe hypertension; the association of hydrochlorothyazide was effective in 5 of 11 such subjects. Thus, enalapril may be safely used as a first stage single therapy in mild or moderate hypertension. PMID- 2562414 TI - [GABA-benzodiazepine receptor complex: therapeutic role of benzodiazepine antagonists]. PMID- 2562415 TI - [Maternal genital herpes: virus isolation during the third trimester of pregnancy and at delivery]. AB - We followed 42 pregnant women at high risk for genital herpes infection. Viral isolation was attempted weekly from the 34th week of pregnancy. One or more episodes of herpes infection developed in 74% of women. The virus was isolated in 28% after 34 weeks and in 22% after 37 weeks. A prophylactic cesarean section was performed in the latter group; only one of these women had a positive viral isolation at delivery. The HSV-2 of virus was found in 83% of cases and HSV-1 in 17%. PMID- 2562416 TI - [The postoperative survival in pulmonary carcinomas depending on the histological type and stage]. AB - The prognostic significance of age, sex, location of the tumor in the various lobes, size, histological type, node metastases, local extent and stage has been studied in a series of 742 surgically resected lung carcinomas. The histological type was a very important prognostic factor: the highest survival was observed in epidermoid carcinomas, followed by adenocarcinomas, anaplastic large cell carcinomas, and anaplastic small cell carcinomas. The stage, as well, except for the adenocarcinoma, bore heavily on the prognosis; however, in small stage I tumours, the postoperative survival was independent from the histological type. The presence of lymph node metastases resulted in an extremely poor survival, except for the epidermoid carcinoma. The size of the tumours, excluding adenocarcinomas, was an important prognostic factor provided lymph node metastases were absent. No significant differences in survival according to the location in different lobes could be ascertained. PMID- 2562417 TI - Bluetongue epidemiology in the Caribbean region: serological and entomological findings from a pilot sentinel system in Trinidad and Tobago. AB - When monitored by the agar gel immunodiffusion test for antibody to bluetongue viruses, a sentinel flock of twenty-five lambs remained seropositive through the year, whereas in a sentinel herd of twenty calves only two individuals seroconverted and these became negative again within 2 months. A light trap operated with the calf herd yielded high numbers of Culicoides insignis Lutz (over 18,000 per trap night) along with C. filariferus Hoffman, C. pusillus Lutz, C. leopoldi Ortiz, C. foxi Ortiz, C. limai Barretto, C. diabolicus Hoffman and C. guyanensis Floch and Abonnenc. Culicoides were trapped at the sheep station which had housed the lambs 3 years following the sentinel study. No virus was isolated from pools of C. insignis, C. filariferus and C. pusillus. Six other species were collected in insufficient numbers to warrant attempted virus isolations. PMID- 2562418 TI - The vector competence of Culex annulirostris, Aedes sagax and Aedes alboannulatus for Murray Valley encephalitis virus at different temperatures. AB - Culex annulirostris Skuse, colonized from Brisbane, Queensland, and Mildura, Victoria, Australia, were effective vectors of Murray Valley encephalitis virus at 20, 27 and 32-35 degrees C with full extrinsic incubation periods of 15, 10 and 4 days respectively. At 20 degrees C, 7-11 days post-infection, transmission by the Mildura colony (0-20%) was less efficient than the Brisbane colony (30 70%) but both were capable of 75-100% transmission after longer extrinsic incubation periods. Discriminant analysis of body and salivary gland titres showed that these were not satisfactory indicators of transmission. Wild-caught Aedes sagax (Skuse) and Cx annulirostris from the Murray Valley showed equal competence, but Aedes alboannulatus (Macquart) was a poor vector. The results provide data on rural amplification of Murray Valley encephalitis virus during spring and suggest that further work on the potential of Ae. sagax as a natural vector is warranted. PMID- 2562419 TI - Rearing temperature influences flavivirus vector competence of mosquitoes. AB - Culex annulirostris Skuse mosquitoes (Brisbane strain) were reared at 20 degrees C or 27 degrees C and the adult females were experimentally infected by feeding Murray Valley encephalitis virus (MVE). They were then maintained (a) in the insectary at 20 degrees C, after rearing at either 20 degrees C or 27 degrees C; (b) at ambient outdoor temperatures, range 12.2-28.9 degrees C, mean 19.6 degrees C; or (c) at 27 degrees C after rearing at 27 degrees C. There was no significant difference in rates of MVE infection or transmission when mosquitoes were reared and maintained constantly at 20 degrees C or 27 degrees C. However, for females kept at reduced temperature (i.e. mean = 19.6 degrees C or 20 degrees C after rearing at 27 degrees C), the infection and transmission rates of MVE were significantly reduced (2 x 8 replicates). This investigation illustrates that vector competence is depressed by decreasing temperatures for adult mosquitoes compared with those they experienced during development. Similar patterns were evident with previously published work on Japanese and St Louis encephalitis, dengue and yellow fever. The process appears to be reversible, i.e. increased temperature raises virus infection and transmission rates. It is concluded that, without incubation at warmer temperatures, flavivirus recovery from overwintering mosquitoes will be negatively biased. PMID- 2562421 TI - Metabolic activation of eugenol by myeloperoxidase and polymorphonuclear leukocytes. AB - Eugenol has recently been associated with the toxic effects of clove cigarettes on human lungs. We have studied the metabolism and adverse effects of eugenol on human polymorphonuclear leukocytes (PMNs). Myeloperoxidase, isolated and purified from human PMNs, catalyzed the oxidation of eugenol to a reactive intermediate which is likely to be a quinone methide. Eosinophil peroxidase, lactoperoxidase, prostaglandin H synthase, horseradish peroxidase, and rat intestinal peroxidase also supported this hydrogen peroxide dependent reaction. Glutathione inhibited the formation of this metabolite, resulting in the formation of glutathione disulfide and a small amount of eugenol-glutathione conjugates. In cellular incubations, phorbol ester stimulated PMNs catalyzed the covalent binding of [3H]eugenol to cellular protein, which was partially inhibitable by azide. Intracellular glutathione levels decreased by 90% over a period of 30 min in phorbol ester stimulated PMNs exposed to 100 microM eugenol compared with decreases of 30% (phorbol ester alone) or 5% (eugenol alone) in control incubations. In addition, eugenol was more cytotoxic to PMNs in the presence of phorbol ester than in its absence, and eugenol inhibited the phorbol ester stimulated oxidative burst in PMNs as reflected by a decrease in oxygen consumption, superoxide formation, and hydrogen peroxide formation. These results suggest that PMNs are capable of activating eugenol to a reactive intermediate and also suggest a mechanism whereby eugenol can potentially interfere with and adversely affect vital PMN functions. PMID- 2562420 TI - The smoke produced from the oxidative pyrolysis of perfluoro polymers: an ESR spin-trapping study. AB - Spin adducts are observed when the unfiltered smoke produced during the aerobic pyrolysis of perfluoro polymers (PFP) is bubbled through a solution of the ESR spin trap alpha-phenyl-N-tert-butylnitrone (PBN). The spin adducts include those from an oxy radical and the fluorine atom, and in addition the spin trap is oxidized to PBNOx. The spin adduct of a chlorine atom also is observed, presumably because of chlorine-containing impurities in the polymers. Tetrafluoroethylene is produced during the pyrolysis of PFP; therefore, we subjected tetrafluoroethylene to the same pyrolysis as was used for PFP, and we observe the same series of spin adducts. We suggest that the spin adducts formed from the whole, unfiltered smoke from PFP smoke may result both from free radicals that form during the pyrolysis of PFP and from the oligomerization of the tetrafluoroethylene produced during the pyrolysis. These spin-trapping results support the theory that reactive free radicals are present in PFP smoke and may contribute to PFP smoke toxicity. PMID- 2562422 TI - Oxygenation by superoxide ion of halogenofluorocarbons (Freons and haloforms) in aprotic solvents. AB - The reactivity of chloro- and bromofluoromethanes (Freons and others) with superoxide ion (O2.-) has been evaluated by cyclic voltammetry in dimethylformamide (DMF). Substitution of fluorine atoms into chloromethanes results in a substantial decrease in their reactivity with O2.- (relative rates: CCl4 much greater than CF4, much greater than CCl4 much greater than FCCl3, CCl4 much greater than F2CCl2, CCl4 much greater than F3CCl, H3CCl much greater than F3CCl, and H2CCl2 much greater than F2CCl2). The bromo derivatives react much faster with O2.- than the corresponding chloro compounds (F3CBr much greater than F3CCl, F2CBr2 much greater than F2CCl2, and FCBr3 much greater than FCCl3). The rates of reaction for HCCl3, HCFCl2, and HCF2Cl are approximately the same, and the reactions appear to have a common path via dehydrochlorination. Reaction stoichiometries, apparent second-order rate constants, and product profiles have been determined for these fluoromethanes and related chlorofluoroethanes. PMID- 2562423 TI - Site-specific DNA damage induced by cobalt(II) ion and hydrogen peroxide: role of singlet oxygen. AB - The effect of Co(II) ion on the reaction of hydrogen peroxide with DNA was investigated by a DNA sequencing technique using 32P-5'-end-labeled DNA fragments obtained from human c-Ha-ras-1 protooncogene. Co(II) induced strong DNA cleavage in the presence of hydrogen peroxide even without alkali treatment. Guanine residues were the most alkali-labile site, and the extent of cleavages at the positions of thymine and cytosine was dependent on the sequence. Adenine residues were relatively resistive. Diethylenetriaminepentaacetic acid, present in excess over Co(II), inhibited DNA cleavage. Singlet oxygen scavengers (dimethylfuran, sodium azide, 1,4-diazabicyclo[2.2.2]octane, dGMP), sulfur compounds (methional, methionine), and superoxide dismutase inhibited DNA cleavage completely. Hydroxyl radical scavengers were not so effective as singlet oxygen scavengers. ESR studies using 2,2,6,6-tetramethyl-4-piperidone as a singlet oxygen trap suggest that Co(II) reacts with hydrogen peroxide to produce singlet oxygen or its equivalent. ESR studies using 5,5-dimethylpyrroline N-oxide (DMPO) showed that the hydroxyl radical adduct of DMPO was also formed. The results suggest that Co(II) ion binds to DNA and subsequently reacts with hydrogen peroxide to produce singlet oxygen and hydroxyl radicals and that singlet oxygen plays a more important role in the DNA damage than hydroxyl free radicals. PMID- 2562424 TI - 2,3-Dithioerythritol, a possible new arsenic antidote. AB - British antilewisite (2,3-dimercaptopropanol; BAL) has long been used as an arsenic antidote, but its therapeutic efficacy is limited by its inherent toxicity. We synthesized two less toxic derivatives of BAL and investigated their potential as antidotes to organic arsenic. The new compounds, 2,3 dithioerythritol (DTE) and 2,2-dimethyl-4-(hydroxymethyl)-1,3-dithiolane (isopropylidene derivative of BAL), react readily with phenyldichloroarsine (PDA) to yield the expected corresponding cyclic 1,3-dithioarsolanes. The BAL derivatives were compared to BAL in terms of their cytotoxicity and their ability to rescue PDA-poisoned mouse lymphoma cells in culture. The dithiolane was not a good antidote in the cultured cell system. In contrast, DTE was less toxic than BAL or DMSA and was superior at improving cell survival in PDA-exposed cells. PMID- 2562425 TI - Studies of intramolecular rearrangements of acyl-linked glucuronides using salicylic acid, flufenamic acid, and (S)- and (R)-benoxaprofen and confirmation of isomerization in acyl-linked delta 9-11-carboxytetrahydrocannabinol glucuronide. AB - NMR and HPLC have been used to investigate the rearrangements of 1-O acylglucuronides in vitro and the occurrence of rearranged isomers in urine. Glucuronides of flufenamic acid, (S)- and (R)-benoxaprofen, salicylic acid, and delta 9-11-carboxytetrahydrocannabinol were synthesized, by use of immobilized enzymes, or purified from urine. Ester-linked isomers of these gluruconides were characterized, and isomers derived from flufenamic acid, (S)-benoxaprofen, and salicylic acid were purified for further study by NMR and HPLC. The positions of the new ester linkages could be identified by two-dimensional NMR. Shifts not only in the resonance of the proton adjacent to the esterified hydroxyl group but also in the resonance of the anomeric proton on carbon 1 of the glucuronic acid moiety could be correlated with the position of each isomeric ester bond. HPLC elution times also correlated with ester position in this small set of samples. The sequences of isomer formation were studied in situ by NMR and also at pH 8 by HPLC. These studies indicate that, for the three cases examined, the C-2 ester is formed first, followed by formation of C-3 and C-4 esters. The purified isomeric esters were found not to re-form the high-energy 1-O-acyl bond. All other rearrangement steps are reversible. In contrast to other glycosides and glycerol esters, no evidence could be found for rearrangements beyond nearest-neighbor hydroxyl groups in glucuronic acid. The sequence of formation and reversibility is consistent with an ortho ester intermediate, as has been proposed for rearrangements of other glycosides.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562426 TI - Oxidations of the carcinogen N-hydroxy-N-(2-fluorenyl)acetamide by enzymatically or chemically generated oxidants of chloride and bromide. AB - The oxidations of the carcinogen N-hydroxy-N-(2-fluorenyl)acetamide (N-OH-2-FAA) via one-electron (1e-) oxidation to equimolar 2-nitrosofluorene (2-NOF) and N acetoxy-2-FAA and via oxidative cleavage to 2-NOF by chemically and myeloperoxidase (MPO)/H2O2 generated oxidants of Cl- and/or Br- were investigated. 2-NOF was determined spectrophotometrically in the reaction mixtures and by HPLC of their extracts; N-acetoxy-2-FAA was determined by HPLC. In the presence of individual or mixed halides at their physiologic concentrations (0.1 M Cl- and/or 0.1 mM Br-) and pH 4-6, MPO/H2O2-catalyzed oxidation of N-OH-2-FAA to 2-NOF via oxidative cleavage was much greater than 1e- oxidation. At the respective pH optima, oxidation was much more rapid with Br- and Br- + Cl- than with Cl-. HOBr or HOCl + Br- oxidized N-OH-2-FAA more rapidly than HOCl, also chiefly via oxidative cleavage. This suggested that, in the presence of MPO/H2O2 + Cl- + Br-, oxidation was due to HOBr from HOCl oxidation of Br- and/or oxidation of Br- by MPO/H2O2. In the presence of taurine (1 or 10 mM), a scavenger of hypohalous acids, MPO/H2O2 catalysis of oxidative cleavage was unaffected with Br-, prevented with Cl-, and partially prevented with Cl- + Br-. These results were linked to N-halotaurine formation since it was found that N-bromotaurine, but not N-chlorotaurine, oxidized N-OH-2-FAA chiefly to 2-NOF. With time N-chlorotaurine and N-bromotaurine appeared to undergo a pH-dependent halide exchange with Br- and Cl-, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562427 TI - Modulation of immune suppression and enhanced tumorigenesis in retrovirus tumor challenged chickens treated with vitamin E. AB - Vitamin E(dl-alpha-tocopherol) dissolved in ethanol with polyethylene glycol as the vehicle and administered by intraperitoneal injections of 0.1 mg/gm body weight at two day intervals, was demonstrated to be a potent immunomodulating reagent in young chickens challenged with avian reticuloendotheliosis virus transformed tumor cells. Vitamin E treatment enhanced the mitogen-induced proliferative responses of spleen cells from age matched, unchallenged chickens; reduced the tumor cell-induced suppression of host splenic lymphocyte mitogen responses; and eliminated tumor cell-induced suppressor cell activity. However, inspite of an improved immune status the vitamin E treated-tumor cell challenged chickens exhibited enhanced tumorigenesis. PMID- 2562428 TI - The relationship between nutritional antioxidants and serum lipid peroxides in cancer patients. AB - A new parameter, the ratio of lipid peroxide and vitamins E and C [LPO/(VE + VC)], has been proposed and used to reflect the balance between lipid peroxidation and antioxidation capability of cancer patients and of healthy human controls. The effects of vitamins E, C, and selenium on the serum LPO level in mice bearing Ascites Hepatomas (H22) have also been examined. The results showed that the average of LPO/(VE + VC) ratios in cancer patients (135 cases) was significantly higher than that of the normal controls (222 cases). The authors suggest that this ratio might be used as one of the parameters for early diagnosis and prognosis of diseases (including cancers) caused by free radicals and lipid peroxides. The results also showed that antioxidants - Se(Na2SeO3, 1mg/kg) or vitamin E (5mg/kg) could markedly decrease the level of serum LPO in the tumor-bearing animals. A smaller dose of VE (1mg/kg) and doses of Vc up to 300mg/kg showed no effect on the serum LPO levels when given separately. However, synergistic effects were observed when any 2 out of 3 or three nutrients were given together. Those with three nutrients significantly lowered the serum LPO level. These antioxidants also inhibited the proliferation of tumour cells. PMID- 2562429 TI - Choice vegetables simultaneously for beta carotene. Vitamin C and fibre. AB - Beta-carotene, vitamin C, and fibre are found mainly in produce, that is, vegetables, fruits and grains. Unfortunately, grains, which have high content of fibre, are not normally consumed whole, and the most fibrous part might actually miss the alimentary tract. Fruits, which have high content of Vitamin C and are usually consumed unrefined, are expensive when not in season. In this article, we examine the possibility of maintaining recommended levels simultaneously of the nutrients mentioned with a short list of more popular vegetables, namely, broccoli, carrot, peas, sweet potato, and spinach. PMID- 2562430 TI - Measurement of ionized cytoplasmic calcium mobilization with the photoprotein aequorin in human polymorphonuclear leukocytes activated by platelet activating factor (PAF). AB - The use of the sensitive photoprotein aequorin as a Ca2+ indicator in human polymorphonuclear leukocytes (PMN) not pretreated with cytochalasin B and stimulated with platelet activating factor (PAF) may help cast more light on the relative importance of intracellular and extracellular Ca2+ in PMN function. PAF elicits Ca2+ mobilization in PMN (resuspended in the presence of 1 mM extracellular Ca2+), in a concentration-dependent manner. The Ca2+ chelator ethyleneglycoltetraacetic acid (EGTA) abolishes Ca2+ mobilization, suggesting that almost all Ca2+ mobilized by PAF derives from the external medium. Aggregation and enzymatic release parallel the Ca2+ mobilization triggered by PAF. In contrast PAF appears to be only a weak stimulus of superoxide anion production (compared to the phorbol ester phorbol 12-myristate 13-acetate (PMA] and leukotriene B4 (LTB4) synthesis (compared to the Ca2+ ionophore A23187). The fact that PAF elicits Ca2+ mobilization, aggregation, secretion and LTB4 generation in human PMN supports the role of this phospholipid as a powerful mediator of physiopathological events involving PMN activation. PMID- 2562431 TI - Receptor antagonism of leukotriene B4 myotropic activity by the 2,6 disubstituted pyridine analog U-75302: characterization on lung parenchyma strips. AB - Leukotriene B4 constricts guinea pig lung parenchyma strips in a concentration dependent manner. The LTB4 structural analog U-75302, 6-(6-[3-hydroxy-1E,5 Z undecadienyl]-2-pyridinyl)-1,5-hexanediol, was a partial agonist in this system with a potency 300-1000 times less than LTB4. U-75302 constricted lung parenchyma strips only at concentrations greater than 0.3 microM. At concentrations lacking agonist activity U-75302 was an effective antagonist, displacing the LTB4 dose response curve. Half-maximal responses required 0.10 microM LTB4 in the presence of 0.3 microM U-75302 and 0.01-0.02 microM LTB4 in its absence. The maximal force of contraction was unaffected at this concentration. Concurrent with antagonism of the myotropic response, U-75302 inhibited the LTB4-dependent release of thromboxane B2 from lung parenchyma. This effect was attributable to receptor antagonism, not enzymatic inhibition of phospholipase, cyclooxygenase, or thromboxane synthase. For instance, 0.3 microM U-75302 did not inhibit thromboxane B2 formation by lung parenchyma stimulated with calcium ionophore A23187 and it did not inhibit thromboxane B2 formation by human platelets stimulated with arachidonic acid. U-75302 selectively antagonized the activity of LTB4 and not other myotropic agonists including the thromboxane A2 mimetic U 46619, LTC4, LTD4, AGEPC, PGF2 alpha, and histamine. Receptor antagonists of leukotriene B4 may have multiple beneficial effects on asthmatic or respiratory disorders. These include (i) direct antagonism of LTB4 myotropic actions; (ii) antagonism of LTB4-dependent mediator release; and (iii) antagonism of LTB4 chemotactic action associated with leukocyte infiltration during anaphylactic late phase reactions. PMID- 2562432 TI - Time-dependent differences in the vasopermeability response to intradermal peptidoleukotrienes. AB - The time course of extravascular albumin accumulation responses elicited by the leukotrienes LTC4, LTD4, LTE4, and histamine in the skin were compared in the conscious guinea pig. During the initial 15-min period, comparison of the dose response curves revealed that histamine produced a much larger increase in extravascular albumin content than any of the leukotrienes. One hour after intradermal injection and at subsequent time intervals, the response to LTD4 had increased in magnitude so that it equaled the response produced by histamine. This was apparent from comparison of the time courses of extravascular albumin accumulation for intermediate doses of LTD4 and histamine and also from comparison of dose-response relationships at 4 h post intradermal injection. In contrast to LTD4, the magnitude of the microvascular permeability responses to LTC4 and LTE4 remained relatively small even over an extended time scale. Although histamine produced a large initial response, this also remained essentially unchanged over a 4-h period. It appears that LTD4 may produce a unique, time-dependent cutaneous microvascular permeability response and measurements over 15-30-min periods may underestimate its activity as a vasopermeability factor. The time-dependent effects of LTD4 on albumin extravasation cannot be ascribed to leukocyte infiltration into the skin since LTC4, LTD4, and LTE4 were entirely without effect in this regard. PMID- 2562433 TI - Effect of 1-oleoyl-2-acetyl-sn-glycerol on inositol lipid metabolism of ascites tumor cells in culture. AB - 1-Oleoyl-2-acetyl-sn-glycerol (OAG), the membrane-permeable analogue of 1,2 diacylglycerol (DAG), which stimulates ascites tumor cell proliferation, was used to study its effect on phosphoinositide metabolism. Culturing of ascites cells labeled with [3H]inositol at low serum concentration in the presence of OAG suppressed the radioactivity level of the inositol phosphates, particularly IP3. Membrane-bound, Ca(2+)- and GTP gamma S-sensitive PI- and PIP2-specific phosphodiesterase (phospholipase C) showed much lower activities in OAG stimulated cells, which could be enhanced by GTP gamma S in these but not in the unstimulated cells. A high susceptibility to Ca2+ of the PI- and PIP2-specific phospholipase C of non-stimulated cells was observed. The PIP-kinase activity was similarly reduced by about 85% in OAG-stimulated cells. These data indicate a negative feedback regulation of the phosphoinositide metabolism mediated by OAG. Reduction in synthesis and degradation of PIP2, which furnishes the two second messengers, DAG and IP3, provides a means of controlling the intracellular level of these molecules, which is important for a balanced proliferation rate. PMID- 2562434 TI - PAF-receptor. 1. 'Cache-oreilles' effect of selected high-potency platelet activating factor (PAF) antagonists. AB - Three-dimensional electrostatic maps were calculated for six potent antagonists of platelet-activating factor (PAF), the antagonists being selected for their apparent structural heterogeneity. The molecules examined were the compact Ginkgolides BN 52020, BN 52021 and BN 52022 (1, 2 and 3), the semi-rigid kadsurenone (4), a flexible synthetic dinor type C furanoid lignan L-652,731 (5a) and the triazolothienobenzodiazepine WEB 2086 (7). Calculation of the electrostatic potential generated around all the above molecules showed the existence of two wells of negative potential or 'cache-oreilles' (ear-muffs), i.e., the isocontours drawn at -10 kcal/mol, located at 180 degrees from each other and separated by a maximum distance of 22-27 A. Except for the synthetic dinor type C furanoid lignan (5a), the molecules also presented a moderate hydrophobic fragment, which constitutes a third point of interaction with the high-affinity binding site in rabbit and human platelets. The findings of the present study allow speculation that this high-affinity acceptor site may be a 'polarized cylinder' with a diameter of 10-12 A. PMID- 2562435 TI - Na+/H+ exchange in PAF-stimulated platelets. AB - In experiments on human platelets, inhibition of Na+/H+ exchange was caused either by equimolar substitution of external Na+ with choline or N-methyl-D glucamine, by decreasing the pHo to 6.8, or by an inhibitor of the antiport 5-(N ethyl-N-isopropyl)amiloride (EIPA). In all these cases a considerable inhibition of PAF-induced platelet aggregation and as a rule a more or less marked decrease in the cytoplasmic Ca2+ signal (quin-2-loaded platelets) occurred. Stimulation by 10(-7) M PAF caused biphasic pHi changes in human platelets loaded with the pH sensitive fluorescent probe BCECF: a small transient decrease, followed by a sustained increase of 0.02 +/- 0.006 pH units, resulted from stimulation of the Na+/H+ exchange. Thrombin (0.1 U/ml) also caused biphasic pHi changes, but the alkalinization step was more pronounced (0.15 +/- 0.03 U). Every means of Na+/H+ exchange inhibition prevented a rise in pHi in stimulated platelets. Activation of the adenylate cyclase system by carbacyclin suppressed the agonist-induced pHi increase. The inhibition of neither cyclooxygenase by 10(-5) M indomethacin nor calmodulin-dependent enzymes by 10(-5) M calmidazolium affected the agonist induced pHi signals. A decrease in temperature from 37 to 24 degrees C caused a considerable increase in the lag phase of the pHi signal induced by tetradecanoyl phorbol acetate (TPA), but did not affect the kinetics of the pHi signal induced by PAF. An inhibitor of protein kinase C (PKC), compound H-7 (60 microM), completely abolished the TPA-induced increase in pHi but caused only a partial inhibition of the pHi signal in about 50% of the experiments with PAF. On the basis of these results the conclusion is drawn that the activation of PKC is not the only pathway for the PAF-induced stimulation of Na+/H+ exchange. The PAF induced pHi rise depended both on the presence of extracellular Ca2+ and on the [Ca2+]i increase. On the other hand, inhibition of Na+/H+ exchange decreased the magnitude of the Ca2+i signal in PAF-induced platelets loaded with quin-2, but did not influence the Ca2+ mobilization from intracellular stores as measured by quin-2 or chlortetracycline in experiments with thrombin-stimulated platelets. We conclude that in PAF-activated platelets some initial increase of [Ca2+]i is essential for Na+/H+ exchange activation while activated antiport potentiates a full-scale Ca2+ influx into the cells. PMID- 2562437 TI - Alkaline phosphatase expression in human cell lines derived from primary hepatomas. AB - Isozyme patterns of alkaline phosphatase (ALP) were electrophoretically examined in human cell lines derived from one hepatoblastoma, five hepatocellular carcinomas (HCCs) and two cholangiocellular carcinomas. Most of the cell lines tested had a liver-type ALP isozyme. In addition, an abnormal ALP isozyme, which was similar to variant ALP, was detected in one hepatoblastoma and two HCC cell lines. One HCC cell line of these variant-like ALP-positive cell lines was alpha fetoprotein (AFP)-negative. These findings suggest that variant-like ALP may be useful for the identification of human hepatoma cell lines, especially in AFP or albumin-negative cell lines. PMID- 2562436 TI - PAF receptor. 2. Quantitative hydrophobic contribution of the agonist's etheroxid chain. AB - In order to undertake a quantitative assessment of the contribution of the hydrophobic effect of the etheroxid chain to agonistic activity, it was necessary to include in the calculated data a highly lipophilic platelet-activating factor (PAF) analogue. The synthesis of such a compound--racemic 1-O-docosyl-2-O acetylglycero-3-phosphocholine (C22 PAF)--is described. The regression analysis performed with data of 14 etheroxid analogues (reviewed by Godfroid et al., 1987), combined with data obtained with C22 PAF, is significant with respect to a parabolic evolution between lipophilicity of the chain and the logarithm of relative platelet stimulation. This result is characteristic of a hydrophobic interaction between the agonist and the PAF receptor. PMID- 2562438 TI - Nuclear retinoic-acid-binding proteins and receptors in retinoic-acid-responsive cell lines. AB - Nuclear retinoic-acid-binding activity and the expression of retinoic acid receptor mRNA (RAR-alpha and RAR-beta) were assayed in the F9 embryonal carcinoma, HeLa, HL-60 promyelocytic leukaemia and S91 melanoma cell lines. A 4 svedberg nuclear retinoic-acid-binding activity was detected in all 4 cell lines, but the levels in the HeLa and HL-60 cells were lower than in the F9 and S91 lines. RAR-alpha mRNA was expressed in all 4 cell lines, although at a very low level in S91 cells. Conversely, RAR-beta mRNA was expressed in S91 cells and, at a lower level, in F9 cells but was undetectable in HeLa and HL-60 cells. RAR beta, transcribed and translated in vitro from the cloned cDNA coding region, sedimented at 4 S and this suggests that the 4-svedberg nuclear retinoic-acid binding activity may represent the retinoic acid receptors. PMID- 2562440 TI - Detection of enteropathogens in diarrhoeal diseases among malnourished Egyptian infant and children. AB - The influence of the Pre-existing malnutrition (PEM) on the severity of diarrhoea as well as the causative organisms was studied on 60 cases. The duration of diarrhoea was prolonged in cases with PEM. The stool purgative rate ranged from 4 15 times/day in PEM infant while it was 3-6 times in well nourished cases (WNC) (P less than 0.05). Also vomiting and dehydration was more marked among PEM cases (52.9% and 32.4% of cases than in WNC cases (31.3% and 12.5% of cases) (P less than 0.05). Rota virus and Candida albicans were the Commonest identified organisms in the stools of the PEM cases, they were detected in 52% and 38.2% of cases respectively while 25% of WNC had rota virus in their stool and non of them had Candida (P less than 0.02). Giardia lamblia was detected in 23.5% and 18.8% of PEM and WNC while 10% of healthy controls had Giardia. Other bacterial enteropathogen were also found less frequently including Salmonella, Shigella, E. coli, Pseudomonas and Campylobacter. There was no statistical difference in the incidence between both groups. Multiple infections were detected in 47% and 18.7% of PEM cases and WNC (P less than 0.05) and correlated with the severity of illness. PMID- 2562439 TI - In vitro proliferation of murine spleen cells: inhibition by monoclonal antibodies to L3T4 and Lyt-2 T cell markers or intracellular cyclic adenosine monophosphate. AB - Proliferation of normal (not immunized intentionally) murine spleen cells was elicited with concanavalin A, supernatant fluid from cultures of EL-4 cells, human recombinant interleukin 2 (IL-2), or a mixture of phorbol ester and calcium ionophore A23187. IL-2-induced proliferation was inhibited by membrane-permeable dibutyryl cyclic adenosine monophosphate (cAMP) or by the adenylate cyclase activator forskolin. Consistent with these observations was the finding that stimulation with IL-2 decreased and forskolin increased the intracellular content of cAMP. IL-2-induced proliferation, as well as that induced by concanavalin A or phorbol-ionophore mixture, was inhibited by monoclonal antibodies specific for L3T4 or Lyt-2 cell surface markers. This inhibition was observed even when antibodies were added several hours after exposure of cells to IL-2. Notably, antibodies did not alter the intracellular content of cAMP. Thus, the experimental data failed to establish a functional linkage between the inhibitory effect of antibodies and the regulatory effect of the adenylate cyclase system. However, our results provide a rational basis for the postulation that antibodies, upon binding to their corresponding ligands, generate a negative signal that interferes with IL-2-induced proliferation. Therefore, L3T4 and Lyt-2 molecules appear to play an important role in the regulation of lymphocyte proliferation. PMID- 2562441 TI - Development of an enzyme linked-immunosorbent assay (ELISA) for the sand fly fever viruses detection. AB - An Enzyme linked-immunosorbent assay (ELISA) was recently established for the detection of sandfly Naples and Sicilian viruses (SFN and SFS) from laboratory infected as well as wild caught sandflies in Egypt. Optimal dilutions of the reactants including the coating antibodies (Rabbit antiserum), detecting antibodies (Mouse antiserum), conjugate and the time used for incubation of the substrate (ABTS) were determined for both SFN and SFS viruses. The ELISA test showed to be highly specific and sensitive except for the SFN virus which cross reacted with Toscana virus. A total of 1582 sandflies, forming 54 pools (each consisted of 2-109 flies) were collected from different governorates in Egypt (North Sinai, South Sinai, Alexandria, Giza, Qualubyia, Sharkiya and Aswan) in the years 1986-1987 and 1988, and tested for virus detection by the SFN-ELISA and SFS-ELISA. Only one pool (from Giza governorate) revirus was detected. Tests were repeated two times and confirmed by the complement fixation and plaque neutralization tests. The established ELISA technique hold great promise as a routine surveillance tool, permitting rapid, simple, sensitive, specific and inexpensive assay for the detection of the sandfly fever viruses. PMID- 2562442 TI - Beneficial effects of ACE inhibitors in severe mitral stenosis. AB - There are several reports of beneficial effects of ACE inhibitors in both primary and secondary pulmonary hypertension. However the effect of ACE inhibitors in mitral stenosis is not documented. The authors report three patients with severe mitral stenosis in whom surgery was delayed. They had initial symptomatic improvement with diuretics and sodium restriction, but had recurrence of their symptoms while on treatment. Enalapril not only relieved their symptoms in particular exertional dyspnoea and haemoptysis but prevented recurrence and improved their effort tolerance without causing excessive fall of blood pressure or impairment of renal function. PMID- 2562443 TI - Coexisting cervical spondylotic myelopathy and bilateral carpal tunnel syndromes. AB - In six patients, operations for bilateral carpal tunnel syndromes (CTS) were performed or were about to be performed without the awareness of the presence of underlying cervical spondylo-stenosis. Only later, when symptoms of myeloradiculopathy were recognized, was the diagnosis confirmed and decompressive laminectomy performed. Because the symptoms of CTS may resemble or be masked and accentuated by the cervical disorder, patients with the presumed diagnosis of bilateral CTS should undergo appropriate critical neurologic, electrodiagnostic, and neuroradiologic (magnetic resonance imaging, computed tomography, myelo computed tomography) assessment. If these guidelines are followed, the radiculopathy caused by cervical pathology will be appropriately recognized and treated, possibly averting the need for carpal tunnel decompression or modifying treatment. PMID- 2562444 TI - Lumbar spondylosis, spondylolisthesis, neurogenic claudication, and L4 radiculopathy. PMID- 2562445 TI - Functional, biochemical, and structural anomalies in platelets of patients with idiopathic scoliosis. AB - Idiopathic scoliosis (IS) is the most common type of lateral curvature of the spine, the etiology of which is unknown. Among the various pathological changes that have been reported in IS are platelet anomalies. Since the platelets are not involved mechanically in the spinal deformity, platelet disorders may reflect a basic cellular pathology and not a secondary change attributed to the spinal curvature. In the present review, a summary of the platelet anomalies in idiopathic scoliosis is presented. The accumulated data indicate the existence of profound functional anomalies in platelets of patients with IS. An attempt is made to clarify the different anomalies and to try to link them together. PMID- 2562446 TI - Selective nerve root block in patient selection for lumbar surgery: surgical results. AB - The cause of lumbar radicular symptoms often remains elusive after standard clinical and radiographic evaluation. Selective nerve root block is a useful test to indicate whether the pain is neural in origin and/or whether nerve root is pain producing in these patients with equivocal clinical and imaging studies. Over 8 years, the author performed selective nerve root blocks in 215 patients. Of this group, 78 patients underwent surgery. Following surgery, 71 patients were available for a minimum 12-month follow-up. The preoperative diagnoses included previously unoperated-upon lumbar disc herniation, previously unoperated-upon spinal canal stenosis, and prior lumbar surgery. The average follow-up was 34 months (range, 12-96 months). Overall, there were 38 good (53%), 16 fair (23%), and 17 poor (24%) surgical results. The results for those patients who had had prior surgery were disappointing (52% poor). These data reaffirm that surgical intervention should only be recommended for previously operated-upon patients with unequivocal findings. PMID- 2562447 TI - Electrophysiological studies in cervical spondylosis. AB - This paper reports a study of 57 patients with cervical spondylosis who underwent nerve conduction velocity (NCV), cervical somatosensory evoked potentials (CSEP), and concentric needle electromyography (EMG) as an aid to diagnosis. The results were analyzed in two groups. In Group I, there were 24 patients with radiological changes of cervical spondylosis in the absence of clear neurological signs. Nine patients in this group had abnormal electrophysiological recordings. In Group II, there were 33 patients with clinical signs, and 22 had abnormal electrophysiological recordings. The CSEP was abnormal in 12 of 14 patients who had evidence of nerve root filling defects on myelography. It is concluded that NCV, CSEP, and concentric needle electromyography are useful tests in distinguishing between root lesions and peripheral entrapment neuropathy. Although the methods available are not sufficiently precise to localize the level involved, they can be used as a screen prior to myelography or enhanced computed tomography (CT) scanning if surgery is proposed. PMID- 2562448 TI - A method for writing enzyme rate equations: application to the estimation of the number and size of key proton families. AB - We develop a method to derive the rate equation for enzyme models that include pH dependent activation. Our presentation is based on a kinetic model recently described for sucrase, the three-key-proton model of Vasseur and coworkers, which considers the existence, in the acid ionization reaction, of two functionally distinct prototropic groups, respectively responsible for either V-type or K-type kinetic effects. In contrast, as concerns the basic ionization reaction, the model conforms to classical concepts of pH-dependent activation, whereby a single proton participates in either V-type or K-type effects but not in both at the same time. Enzymes with more than three key protons have been described, indicating that, rather than isolated protons, groups of protons should be considered, and therefore the model can be better described as a three-proton family model, where a proton family is defined as one or several protons that are gained or lost as a block and perform the same kinetic function. The resulting model is treated here as a useful framework upon which other models can be built. To facilitate the writing of the rate equations, we define two new entities: (1) intralevel coefficients, which describe the various combinations of the enzyme with either the substrate(s), the allosteric effector(s), or both at a given protonation level, and (2) interlevel coefficients, which describe the interplay between the various protonation levels. The resulting rate equation can be used in a global fit procedure permitting in a single computer run the estimation of (1) the entire set of dissociation and microscopic ionization constants of the model, (2) the number and kinetic function of proton families characterizing the enzyme under consideration, and (3) the number of key protons constituting each family, which is derived from the derivatives of the kinetic parameters, Vm/Km, Vm, and Km. PMID- 2562449 TI - Dynamics of the Adair model for ligand-receptor binding. AB - The dynamics of the Adair model for ligand-receptor binding involving the case with interacting receptors is investigated. Using the methods of formal reaction kinetics, the existence, uniqueness, and asymptotic stability of equilibria within the model are demonstrated. The approximate solutions to the Adair model are found. PMID- 2562450 TI - Quantitation by fast-atom bombardment/mass-analysed ion kinetic energy spectrometry: kinetic analysis of cyclic nucleotide phosphodiesterase activity. AB - Quantitation of cyclic nucleotide phosphodiesterase activity by means of fast atom bombardment (FAB) mass spectrometry with mass-analysed ion kinetic energy (MIKE) spectrum scanning is described. Characteristic peaks of the substrate, cyclic AMP, and product, AMP, were identified in positive-ion FAB mass spectra and MIKE scans of the protonated molecules. By spiking enzyme incubates with known quantities of cyclic AMP and AMP and measuring peak heights in the MIKE spectra of both spiked and unspiked samples, the concentrations of cyclic AMP and AMP in solution at the end of a series of enzyme incubations have been estimated. From the data obtained the Km and Vmax of the enzymes were calculated as 181 microM and 28.6 nmol/min respectively, showing excellent agreement with values of the Michaelis constant, Km = 205 microM and the maximum velocity Vmax = 33.2 nmol/min obtained by radioactive assay. PMID- 2562451 TI - Philanthotoxins. A mass spectrometric investigation. AB - A method employing liquid secondary-ion mass spectrometry (SIMS) in conjunction with metastable-ion measurements (linked scanning at constant B/E) to obtain sequence-specific information for three synthetic polyamine isomers was developed. The normal liquid SIMS spectra gave molecular weight information, but important sequence ions were of low intensity or obscured by the background. The metastable-ion spectra contained important fragment ions in particular due to cleavage along the polyamine chain. One of the three synthetic isomers was identical with a toxin present in the venom of the digger wasp. In conjunction with nuclear magnetic resonance spectroscopic studies, this should be a powerful method for the structural characterization of other closely related toxins present in the venom of this wasp. PMID- 2562452 TI - Blood concentrations of thirteen vitamins in cirrhotic patients. AB - To investigate the nutritional status in the presence of liver disease, blood concentrations of various vitamins were measured in 26 patients with histologically verified cirrhosis and in 45 noncirrhotic patients who served as the controls. Plasma concentrations of vitamin A in cirrhotic patients (16.7 +/- 7.9 micrograms/dl) were lower than in noncirrhotic patients (37.1 +/- 13.1 micrograms/dl), the difference being statistically significant (p less than 0.01). However, the blood concentrations of 12 other vitamins in the cirrhotic patients did not differ significantly from findings in the controls. Serum thyroxine-binding prealbumin and retinol-binding protein (RBP) were also significantly decreased in cirrhotic patients. Elevated vitamin A:RBP molar ratio, mean 1.58, in the cirrhotic patients suggested that the low vitamin A level reflected a low production of RBP in the liver. There were no statistically significant differences in blood levels of 13 vitamins when data on cirrhotic patients, with or without a primary hepatocellular carcinoma, were compared. PMID- 2562453 TI - Active sensitisation modifies beta-adrenoceptor reactivity in guinea-pig trachea. AB - The occurrence of beta-adrenoceptor desensitisation in clinical experience, after long term therapy with specific beta 2-agonists, is still an open question, even if this phenomenon is easily observed in different experimental models. Since the majority of these experiments have been performed in normal animals, we investigated the possible occurrence of beta-adrenoceptor desensitisation in the ovalbumin actively sensitised guinea-pig model of experimental asthma. The isoproterenol concentration-response curves performed in pilocarpine contracted guinea-pig trachea in vitro were shifted to the right by the beta-adrenoceptor desensitisation procedure, which was achieved by the in vitro administration of isoproterenol (10(-5) M x 2 times x 20 min each) both in normal and ovalbumin sensitised tissues. The same desensitisation procedure markedly affected epinephrine-relaxing capacity in both normal and ovalbumin actively sensitised guinea-pig tracheae. However, the ovalbumin sensitised tissue seemed to be more sensitive than normal to the specific beta 2-agonist procaterol; in parallel the beta 2-mediated relaxation was more impaired by isoproterenol-induced beta adrenoceptor down regulation in ovalbumin sensitised trachea when compared to normal. Similar results have been obtained using salbutamol as the beta 2 adrenoceptor desensitising agent. The changes in beta-adrenoceptor reactivity between normal and ovalbumin sensitised guinea-pig tracheae seemed to depend on the active sensitisation process. No difference in the degree of beta adrenoceptor down regulation was observed in passively ovalbumin sensitised guinea-pig trachea as compared to normal. These data suggest that, in this model of experimental asthma, beta-adrenoceptor reactivity is in some way modified and that this phenomenon might contribute to the genesis of asthma. PMID- 2562454 TI - Classification of beta-adrenoceptors in ferret tracheal smooth muscle by pharmacological responses. AB - Beta-adrenoceptors in ferret tracheal smooth muscle were classified into beta 1 or beta 2 by determination of (i) the potency rank order for isoprenaline, noradrenaline and adrenaline, and (ii) apparent pA2-values for a specific beta 1 antagonist (practolol) and a specific beta 2-antagonist (ICI 118.551) and (iii) concentration-inhibition curves for procaterol, a beta-agonist showing a biphasic concentration-response curve in organs with a mixed beta 1 and beta 2 adrenoceptor population. We used in vitro tracheal rings and assessed the responses to beta-adrenoceptor agonists as inhibition of phasic contractions elicited by electrical field stimulation (2 Hz for 20 s) or relaxation of tonic contractions elicited by acetylcholine (0.5 microM). The potency rank order for the agonists was isoprenaline greater than noradrenaline approximately adrenaline indicating action through beta 1-receptors. Apparent pA2-values for the antagonists were for 1, 3 and 10 microM practolol 6.0 (SE 0.27), 6.1 (SE 0.21) and 6.3 (SE 0.03), respectively. Apparent pA2 for 0.1, 1 and 10 microM ICI 118.551 were 6.8 (SE 0.25), 6.7 (SE 0.12) and 6.1 (SE 0.14), respectively. These values agree well with published pA2-values for the action of these drugs on beta 1-adrenoceptors. However, the Schild plot for ICI 118.551 was alinear indicating a possible heterogeneity of the beta-adrenoceptors in ferret trachea. The concentration-response curve for procaterol showed the biphasic form typical for organs with a mixed beta 1 and beta 2 population. However, the lower part of the curve, reflecting stimulation of beta 2-adrenoceptors reached only 17.5% (SE 1.4) of the maximally achieved inhibition.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562455 TI - Effects of adrenergic agonists and adenosine on cholinergic neurotransmission in human tracheal smooth muscle. AB - There is only limited information available on the prejunctional regulation of acetylcholine (ACh) release from cholinergic nerves in human airway smooth muscle. Stimulation of cholinergic nerves in fresh postmortem tracheal muscle strips with electrical field stimulation (EFS) causes reproducible contractions. We have studied the effect on contractile responses of: 1) The alpha 2 adrenoceptor agonist effect of noradrenaline (NA, 0.1-30 microM) and clonidine (10 nM-30 microM), in the presence of 1 microM propranolol and prazosin +/- idazoxan (0.1 microM); 2) The beta-adrenoceptor agonist effect of fenoterol (FEN) and isoprenaline (ISO, 1 nM-30 microM) +/- ICI 118,551 (10 nM), comparing EFS responses to comparable responses to exogenous ACh; 3) The A1 and A2 adenosine receptor agonists effects of L-PIA and NECA (1 nM-10 microM). NA caused a concentration-dependent depression of the cholinergic frequency-response curve. However responses at 5 Hz were not modified by the addition of idazoxan. Similarly clonidine did not reduce contractile responses. The concentrations of isoprenaline (56 nM) and fenoterol (165 nM) required to inhibit EFS (5 Hz) by 50% (IC50) were significantly less than those required to inhibit closely matched ACh responses to a comparable degree (ISO = 117 and FEN = 304 nM), and the maximum inhibition of EFS was greater. Following isoprenaline and the beta 2-antagonist ICI 118,551 the IC50's for EFS and ACh were not different. NECA and PIA had no effect on cholinergic EFS. We conclude that a prejunctional beta 2 receptor may be present on cholinergic nerves in post-mortem tracheal smooth muscle but no evidence for alpha 2-adrenoceptor or adenosine-receptor regulation was obtained. PMID- 2562456 TI - Ca-independent augmentation of endplate potential during repetitive stimulation. AB - To elucidate the role of intracellular Ca ions in neuromuscular transmission, we investigated using frog neuromuscular preparations the effect of 1,2-bis(o aminophenoxy)ethane- N,N,N',N'-tetraacetic acid (BAPTA) on frequency augmentation potentiation (formerly frequency facilitation), which have been shown to be useful in identifying the sites of actions of various cholinergic agents. Buffering of intracellular Ca ions by BAPTA was found to suppress only Ca dependent component of frequency augmentation-potentiation (mo); its stimulation frequency dependent factor (k) remained unaffected. Depression by BAPTA treatment of the short-term facilitation of endplate potential (EPP) was the same in both the resting and augmented states. However, the effect of BAPTA was antagonized, but only partially, with the administration of Ca-ionophore, A23187. These suggest that Ca-buffering capability of the intracellularly loaded BAPTA was maintained during low frequency repetitive stimulation. In addition, the post tetanic potentiation of miniature EPP frequency in Ca-free EGTA Ringer was practically unaffected with BAPTA treatment. Undoubtedly Ca ions were essential for transmitter release and short-term facilitation, but Ca ions were not solely responsible for all the changes of transmitter release. The contribution of transmitter mobilization to maintaining the synaptic transmission during repetitive stimulation should be reevaluated. PMID- 2562457 TI - Convergence of interests in antibiotic-induced neuromuscular blockade. PMID- 2562458 TI - How similar are the actions of crotoxin and beta-bungarotoxin? PMID- 2562459 TI - Crotoxin, half-century of investigations on a phospholipase A2 neurotoxin. AB - Crotoxin, the major toxic component of the South American rattlesnake, Crotalus durissus terrificus, is a neurotoxic phospholipase A2 which exerts its pathophysiological action by blocking the neuromuscular transmission. Crotoxin acts primarily by altering the acetylcholine release from the nerves terminals through a mechanism which has not yet been elucidated. It also acts on postsynaptic membranes by stabilizing the acetylcholine receptor in an inactive conformation very similar to the desensitized state. Crotoxin is made of two dissimilar subunits: a basic and weakly toxic phospholipase A2 component-B, and an acidic and non toxic component-A which does not possess any enzymatic activity. Binding experiments showed that crotoxin subunits dissociate when crotoxin interacts with biological membranes: Component-B binds, whereas component-A appears free in solution. The phospholipase A2 subunit binds in a non saturable, non specific manner, on any kind of biological membranes, whereas in the presence of component-A it interacts only with a limited number of high affinity binding sites present on synaptic membranes but not on erythrocyte membranes. Although the target site (acceptor) of crotoxin has not yet been formally identified, binding experiments carried out with small unilamellar phospholipid vesicles of different compositions indicate that some negatively charged phospholipids like mono and diphosphoinositide phosphates might be an important component of crotoxin acceptor site. Crotoxin is in fact a mixture of several isoforms which have very similar but not identical polypeptide sequences. An individual Crotalus durissus terrificus snake is able to synthesize several crotoxin isoforms which may result of the expression of several isogenes and/or of post-translational events. When compared in quantitative manner, the crotoxin isoforms slightly but significantly differ in their enzymatic and pharmacological properties. Finally, immunochemical investigations carried out with polyclonal antibodies prepared against both crotoxin subunits, showed that non precipitating anti-component-B- antibodies (Fab) inhibit the phospholipase A2 activity of crotoxin and neutralize its lethal potency, suggesting that the catalytic and toxic sites of crotoxin are closely related. PMID- 2562460 TI - Effect of repeated irradiation of low-power He-Ne laser in pain relief from postherpetic neuralgia. AB - In order to investigate the efficacy of repeated irradiation of low-power helium neon laser in pain relief, we irradiated 36 outpatients suffering from postherpetic neuralgia. Each patient underwent 20 trials of irradiation on several points around the painful area at a frequency of 2 or 3 times a week. The efficacy of the laser at the end of 20 trials was noticed on 88.9%, and the degree of pain relief was 55.3%, which correlated with the number of trials. These results suggest that the irradiation of He-Ne laser is an effective and safe treatment for postherpetic neuralgia. PMID- 2562461 TI - Postherpetic neuralgia: clinical experience with a conservative treatment. AB - Ninety-seven consecutive cases of postherpetic neuralgia (PHN) were retrospectively reviewed. Patients comprised 49 women and 48 men with a mean age of 71.6 years. The most common painful locations were the chest and upper back (34%), abdomen and lower back (25.2%), and face (20.2%). Burning pain was the most common type of pain (61.3%). Lancinating pain was reported by 40% and throbbing pain by 22.6%. Treatments included drugs (mainly tricyclic antidepressant, anticonvulsant, and neuroleptic drugs), transcutaneous electrical nerve stimulation (TENS), and dry needling of muscles in the affected dermatomes. Positive response to treatment occurred in 18.5% of the patients after one visit. In 9.3% of the patients, the pain still could not be controlled after 10 visits of 2-week intervals. TENS proved to be effective in patients whose skin sensation was preserved. It was concluded that in most PHN cases, pain can be effectively controlled by conservative noninvasive therapy. PMID- 2562462 TI - Immunohistochemical study of a case of malignant mullerian mixed tumor in comparison with the activity of normal uterine tissue. AB - A case of malignant Mullerian mixed tumor of the uterus, exhibiting a histology of heterologous osteosarcomatous differentiation, is presented. Special emphasis is placed on the characteristic immunohistochemical reactivity of the tumor tissue in comparison with that of normal uterine tissue in the proliferative phase. Keratin, cytokeratin, epithelial membrane antigen (EMA), carcinoembryonic antigen (CEA), and human chorionic gonadotropin (HCG) were found only in the carcinomatous element. However, neuron specific enolase (NSE), S-100 protein (S 100) and vimentin were identified in almost all tumor tissue elements. Desmin and actin were not stained in any elements. Myoglobin was only detected weakly in the squamous carcinomatous element. Undifferentiated cell element, composed of small, round, spindle, or polygonal cells showed positive reactions to NSE and S 100, but not to any other antibodies. As compared to the reactivities of the normal proliferative endometrium, the glandular epithelial cells were positive with NSE, S 100, vimentin and CEA, but the stromal cells were only positive with vimentin. Such a multitudinous and concomitant expression of antigenicity to the different tumor elements indicates a close relationship to its mesodermal Mullerian origin, and NSE, S 100 and vimentin might be most adequate indicators of these types of tumors. PMID- 2562463 TI - Effects of glia-conditioned medium on primary cultures of central neurons. AB - The effects of glia conditioned media on the survival and differentiation of embryonic neuron cultures of the Central Nervous System is described. We established glial cultures of peritumoral areas and the culture medium was changed serially and collected as glia conditioned medium (GCM). Neuron enriched cultures plated on poly-L-lysine coated Petri dishes and after 4 days in vitro, the neuronal cultures were treated for the inhibition of glial cells. We established two groups of neuronal cultures that were respectively exposed to the GCM and to conventional culture medium. We evaluated the survival and differentiation of neuronal population in each group of cultures by contrast phase microscopy and cellular uptake of Horse Radish Peroxidase. The cell cultures exposed to GCM showed a survival during the in vitro stages and an organization and differentiation pattern of mature neurons larger than the control cultures. This fact suggests that the glial cultures produce a diffusible neurotrophic factor that influences the neuronal response in vitro. PMID- 2562464 TI - Nuclear bodies in the normal and hyperfunctional human adrenal cortex. AB - Adrenal pieces obtained from six female patients, three without increased adrenocortical function and three with Cushing's disease, showed, in all adrenal cortex zones, cells containing simple and complex nuclear bodies. The simple nuclear bodies were spherical or ovoid and had a filamentous structure surrounded by a clear halo. Complex nuclear bodies were more numerous and heterogeneous in patients with adrenal pathology, and they were spherical with a proteinaceous filamentous capsule surrounding a core; the core was granular, filamentous or a mixture of granular and filamentous material, sometimes with a reticular or concentric arrangement. Some bodies showed vacuolar or multilocular aspect, and others had a close relationship with the nucleolus or appeared near the interchromatin granules. The meaning of adrenal nuclear bodies is discussed as well as their relationship with ACTH stimulation. PMID- 2562465 TI - The histological and the histopathological pattern of conjunctival rhinosporidiosis associated with papillomavirus infection. AB - The present study describes for the first time, the clinical, light and electron microscopic findings of two cases of conjunctival rhinosporidiosis. One was with concurrent infection of papillomavirus. Investigations at the ultrastructural level have provided additional information on the development of Rhinosporidium seeberi and would suggest that the formation of the wall of this organism is a continuous morphological and biochemical spectrum throughout its cytological maturation. The current observation on the wall formation is probably a modification of the classical pattern as an environmental protection carried out by the fungus against the virus. In contradistinction to the usual histopathological picture of rhinosporidiosis, the case with the viral infection lacked the characteristic marked inflammatory reaction. This finding, together with the relatively short interval of the frequent recurrences of this lesion, have led us to postulate the presence of a localised acquired immune deficiency state. It is possible that this local immune deficiency may be caused by an immunosuppression mechanism. This is probably mediated by papillomavirus and/or due to the weak antigenicity of the host virus-infected cells which contain only copies of viral DNA in an unintegrated form. PMID- 2562466 TI - Liver-cell adenoma in an epileptic man on barbiturates. AB - The case of a 19-year-old epileptic man with a solitary hepatic adenoma is described. The tumor was 9 x 8.5 x 6.5 cm in size and microscopically consisted of cells similar in appearance to non-neoplastic hepatocytes, arranged in cords with slit-like sinusoids interposed. Bile ducts and portal tracts were conspicuously absent. Our patient was on antiepileptic drugs, among them phenobarbital which experimentally produces liver cell tumors in mice and rats. PMID- 2562467 TI - Immunoelectron microscopy of human retroviruses. AB - The ultrastructural detection and identification of human retroviruses--HTLV (human T-cell lymphotropic virus) and HIV (human immunodeficiency virus)--have become an everyday task for pathologists and virologists as well as for cell and molecular biologists. The development of better and conventionally available immunocytochemical techniques, such as pre- or postembedding immunocytochemical methods, cryofixation-variants and low temperature embeddings, have made it possible to use them in this field. With the help of these methods the structural proteins of HTLV-I and HIV have been identified in infected cells. The virus assembly at the cell membrane has also been described in detail. Using these methods the incorporation of human transplantation antigens into the envelope of these viruses can be followed. Future studies should establish the pathological significance of this process. PMID- 2562468 TI - The emergence of potent and selective peptide leukotriene receptor antagonists. AB - Leukotriene (LT) C4, LTD4 and LTE4 collectively comprise the constituents of slow reacting substance of anaphylaxis. Based on their well-documented physiology, and a substantial body of circumstantial evidence, it has been hypothesized that they may be etiologic in allergic diseases, including asthma. Using various chemical approaches, a variety of chemically distinct, highly potent and selective LT antagonists have been disclosed including SKF 104,353, ICI 198,615, L 660,711 and WY 48,252. All are, or will soon, enter clinical trials for asthma. These compounds should provide a viable test for the hypothesis that LTs are etiologic in asthma. The complexity of the disease suggests that clinical expectations for these compounds, or any single entity, should be moderate. PMID- 2562469 TI - Human airway smooth muscle in cell culture: control of the intracellular calcium store. AB - The release of intracellularly stored calcium (Ca2+) contributes to the rise in cytoplasmic Ca2+ concentration during agonist-induced activation of airway smooth muscle. We describe a novel preparation--human airway smooth muscle cells cultured in monolayers--which has enabled the investigation of the identity and the function of this intracellular Ca2+ store. Cells, enzymatically dispersed from surgically resected human bronchi, were cultured in monolayers and confirmed as smooth muscle by positive immunocytochemical staining for actin and myosin. The release of intracellularly stored Ca2+ in response to bronchoconstrictor agonists, histamine and carbachol, was demonstrated by stimulation of Ca2+ efflux from 45Ca-labelled cells. The technique of permeabilisation of the plasmalemmal membrane by saponin allowed the measurement of the Ca2+ content of the intracellular store in 45Ca-labelled cells. Uptake of Ca2+ by the store was energy-dependent and was enhanced by cyclic AMP. The effects of inhibitors of sarcoplasmic reticulum and mitochondrial function on Ca2+ uptake identified the store as sarcoplasmic reticulum. Inositol 1,4,5-trisphosphate released stored Ca2+ in a time and dose-dependent manner, supporting its putative role as an intracellular messenger for Ca2+ release in human airway smooth muscle. PMID- 2562470 TI - Modulation of cholinergic neurotransmission by vasoactive intestinal peptide and peptide histidine isoleucine in guinea-pig tracheal smooth muscle. AB - There is increasing evidence that vasoactive intestinal peptide (VIP) and peptide histidine isoleucine (PHI) are non-adrenergic, non-cholinergic (NANC) inhibitory neurotransmitters in airway smooth muscle. The possibility that VIP and PHI may also have neuromodulatory effects on excitatory responses, mediated by cholinergic nerves, to electrical field stimulation (EFS) was studied in guinea pig isolated trachea. VIP (0.5 nM) pre-junctionally, inhibited the release of acetylcholine (ACh), whereas post-junctionally, responses to methacholine (MCh) were enhanced. At a maximum relaxant concentration (100 nM), VIP inhibited cholinergic neurotransmission both pre- and post-junctionally. Similarly, PHI (30 nM) inhibited neuronal ACh release, but enhanced transmitter action post junctionally. At 3 microM, PHI inhibited ACh release. VIP- and PHI-induced inhibition of EFS was not affected by methysergide, pyrilamine, naloxone, phentolamine and propranolol. These data suggest that, in airway smooth muscle VIP and PHI may modulate cholinergic transmission via specific receptors. PMID- 2562471 TI - Inflammatory passage of plasma macromolecules into airway wall and lumen. AB - Anaesthetised guinea-pigs received tracer macromolecules 70-340 kDa intravenously and their erythrocytes were labelled in vivo with 99mTc. Superfusion of tracheal mucosa (via oral catheter) with control solutions and inflammatory agents, was followed by sampling of tracheal surface liquids and tracheal tissue. Under baseline conditions no 125I-fibrinogen (340 kDa) and minimal amounts of erythrocytes, 131I-albumin (70 kDa), and FITC-D (150 kDa) were found in tracheal lavage fluids. Undisturbed baseline conditions with negligible leakage of plasma into airway tissue and lumen were thus obtained with the present provocation and sampling techniques. Superfusion during 2 min with bradykinin 2-10 nmol, histamine 2-8 nmol, capsaicin 0.1-0.4 nmol, PAF 4-8 nmol, ovalbumin 3-6 pmol (in sensitised animals) produced, within 1-10 min, a significant and dose-dependent accumulation of plasma in tracheal tissue and lavage fluids. PAF also induced a late phase plasma leakage response at 5 h. At 10 min PAF given intra-arterially produced a similar leakage into the tissue but less into the lumen compared to topical PAF. Intravenous PAF produced additional effects such as pulmonary oedema. Carbachol 8-16 nmol had only minimal effects on 'leakage' but produced severe bronchoconstriction. Toluene diisocyanate (0.003-0.03 microliter) produced dose-dependent and very sustained (17 h) plasma leakage. Recovery of plasma tracers in airway tissue and surface liquids, respectively, was significantly correlated. As examined with capsaicin, absorption of luminal macromolecules increased only slightly during the exudation process. It is suggested that the consistent inflammatory stimulus-induced passage of plasma into the lumen is a consequence of a load on the basal side of the epithelium induced by the extravasated plasma and its derived peptides. An increased interstitial pressure may transiently separate many epithelial cells allowing a mainly uni-directional almost unrestricted flow of large solutes into the lumen. PMID- 2562472 TI - The protective effects of intravenous theophylline and enprofylline against histamine- and adenosine 5'-monophosphate-provoked bronchoconstriction: implications for the mechanisms of action of xanthine derivatives in asthma. AB - The contribution of adenosine antagonism to the anti-bronchoconstrictor effect of xanthines was examined in 7 atopic asthmatic subjects. On separate occasions the effect of intravenous infusions of theophylline, enprofylline and saline placebo was observed on bronchoconstriction provoked by increasing inhaled concentrations of histamine and adenosine 5'-monophosphate (AMP). Airway calibre was followed as the maximum expiratory flow at 70% below total lung capacity (Vmax30) and FEV1. Both active drugs produced a similar 6-8% increase in FEV1 and 28-41% increase in Vmax30 at steady state plasma concentrations of theophylline 11 mg/ml and enprofylline 3 mg/ml respectively. During the placebo infusion histamine and AMP caused dose-related reductions in both indices of airway calibre, with AMP being approximately 10 times less potent than histamine in molar terms. Theophylline and enprofylline produced similar protection against histamine induced reductions in FEV1 and Vmax30. Whereas enprofylline afforded a similar degree of protection against the airway effect of AMP and histamine, theophylline produced significantly greater protection against AMP whether airway calibre was assessed as Vmax30 or FEV1. The differential effects of theophylline and enprofylline against AMP, but not histamine or baseline airway calibre suggest that adenosine antagonism has little role to play in the acute bronchodilator actions of xanthines. PMID- 2562473 TI - The effect of inhaled ouabain on bronchomotor tone and histamine responsiveness in asthmatic patients. AB - We have studied the effect of inhaled ouabain on resting bronchomotor tone and histamine responsiveness in eight asthmatic patients in an attempt to clarify the role of Na/K ATPase in airway reactivity. Doses ranged from 50 micrograms to 5000 micrograms and were given in a placebo controlled, double blind manner. Baseline FEV1 was recorded prior to inhalation and at intervals up to 30 min thereafter. At 30 min, a histamine provocation test was performed and results expressed as that concentration producing a 20% fall in FEV1. Maximum fall in FEV1 was not significantly different between placebo or any dose of ouabain, ranging from 4.3% 8.2% (p greater than 0.06) nor was histamine reactivity altered significantly (geometric mean range 0.17 mg-0.29 mg [p greater than 0.2]). Unlike animal studies therefore, we have been unable to demonstrate any effect of Na/K ATPase inhibition on airway reactivity in vivo. PMID- 2562474 TI - Nedocromil sodium and sensory nerves in the dog lung. AB - The effect of nedocromil sodium on the main sensory nerve types in the dog lung has been studied. Nedocromil sodium (0.1, 1.0 and 10 mg/kg i.v.) did not stimulate or inhibit the discharge pattern of pulmonary stretch receptors, rapidly adapting irritant receptors or pulmonary C-fibre endings. Nedocromil sodium 5 micrograms/kg given into the aortic arch did, however, stimulate bronchial C-fibre endings. These endings were also stimulated when the drug was given by aerosol. The possibility that nedocromil sodium suppresses cough in the dog by stimulation of bronchial C-fibre endings is discussed. PMID- 2562475 TI - Comparison of in vitro drug responses in airways of atopic dogs with and without in vivo airway hyperresponsiveness. AB - For comparison with previous studies in greyhounds and in the Basenji-Greyhound dog model of asthma (BG), basenji dogs were studied under identical conditions with respect to airway responsiveness to inhaled methacholine, cutaneous responses to intradermal antigen injection, and the sensitivity of isolated trachealis muscle to methacholine and isoproterenol. The relaxant effect of isoproterenol was assessed in trachealis muscle precontracted with methacholine (ED50). The basenji dogs resembled the BG dogs in that they showed multiple positive skin tests. Further, trachealis muscle showed a markedly reduced sensitivity to methacholine (pD2 6.64 +/- 0.10) (+/- S.E.) in vitro. However, basenji dogs resembled the greyhounds in requiring high concentrations of methacholine aerosols to produce a 2-fold increase in pulmonary resistance (1.68 mg/ml +/- 1.21). Thus, there were no significant correlations between sensitivity to methacholine in vitro and airway responsiveness to methacholine in vivo; however, the reduced sensitivity to methacholine in vitro in both basenji and BG dogs may be related to the marked atopy characteristic of both groups. In vitro sensitivity to isoproterenol was correlated (r = 0.82) with the concentration of methacholine needed to elicit the test contraction, but isoproterenol sensitivity in BG dogs was significantly less (p = .0027) than that predicted by the common regression line. This deficit in beta adrenergic function in trachealis muscle unrelated to atopy may be important in the in vivo airway hyperresponsiveness of BG dogs. PMID- 2562476 TI - The teratogenic effect of forskolin on cardiovascular development in the chick embryo. AB - The cardiovascular teratogenicity and embryotoxicity of forskolin, a potent activator of adenylate cyclase, was studied in the chick embryo. The drug was topically applied to the surface of the chorioallantoic membrane in the vicinity of the embryonic heart on the 4th day of incubation (Hamburger-Hamilton developmental stage 24). Cardiovascular malformations were induced in 51% of embryos treated with forskolin doses larger than 1 x 10(-8) mol/egg. Major malformations included remnant of the left 4th aortic arch and ventricular septal defect. These results indicate that forskolin induces cardiovascular malformations in the chick embryo and suggest that increased levels of cyclic 3',5'-adenosine monophosphate produced by forskolin may be related to the malformations observed. PMID- 2562477 TI - Discovery of the non-A, non-B hepatitis virus: the end of the beginning or the beginning of the end. PMID- 2562478 TI - Conversion of ABO blood groups. AB - Progress is being made toward producing erythrocytes similar to native group O cells from A and B donors. Blood group A and B antigens are known to be carbohydrate in nature. The antigenicity is conferred by different terminal sugars. Removal of these sugars by specific exoglycosidases produces the H antigenic structure that is the determinant found on group O cells. Conditions have been developed that allow for the removal of these antigens while maintaining the metabolic and membrane viability of the red cell. Following successful autologous transfusions with gibbons, appropriately treated human group B erythrocytes are now being used in preclinical studies with normal healthy human volunteers. Results indicate that such treated cells have normal in vivo life spans in both group A and O recipients. However, the latter exhibit a transitory increase in anti-B antibody titer, the significance of which is not yet known. Similar exoglycosidic treatment of group A erythrocytes does not remove all serologically detectable A antigens. This is probably due to the presence of a second internal A antigenic site adjacent to the usual terminal A antigen on some structures. Several approaches are being used to address this problem, including projected treatment with a combination of pertinent exoglycosidases and a search for an endoglycosidase that will cleave the polysaccharide chain at a site upstream from both A antigens. PMID- 2562479 TI - The trophic effects of gastrin on fundic neuroendocrine cells of the rat stomach. AB - The histomorphological effect of multidose administration of 6 mg/kg pentagastrin b.d. for 5 weeks, and 1000 mg/kg sodium bicarbonate b.d. for 13 weeks, on the rat fundic mucosa has been examined. Sodium bicarbonate induced a significant hypergastrinaemia (plasma gastrin concentrations were 370.5 pg/ml in the control versus 642.6 pg/ml in sodium bicarbonate-treated rats after 13 weeks, P less than 0.01). Both treatment regimens induced fundic neuroendocrine cell hyperplasia. The cellular proliferation that occurred following hypergastrinaemia of endogenous or exogenous origin suggests that systemic gastrin concentrations play a major role in the control of fundic neuroendocrine cell populations. PMID- 2562480 TI - Measurement of left ventricular ejection fraction and volumes with three dimensional reconstructed transesophageal ultrasound scans: comparison to radionuclide and thermal dilution measurements. AB - A transesophageal, ultrasonic cardiac imaging probe was built that incorporated a mechanism for changing the angle of the imaging plane of a conventional phased array in a precise and known manner. This probe was used to acquire an angular spatial sequence of two-dimensional images of the left ventricular cavity over a series of cardiac cycles by sweeping the imaging plane through it stepwise. The endocardial borders of these images were manually outlined off-line and the application of a three-dimensional reconstruction algorithm was then used to compute the left ventricular end-diastolic and end-systolic volumes and ejection fraction. A study was conducted with seven anesthesized dogs to compare ultrasonic determinations by this method with determinations and measurements made using radionuclide and thermal dilution methods. Comparison of 33 ejection fractions, measured by the ultrasonic volume method and by the gated blood pool radionuclide approach, yielded a correlation coefficient of 0.87 and a standard error of the estimate of 5.7% measured over a range of 10% to 58% (average, 40%). Comparison of the ultrasonically measured volumes with those calculated from stroke volume (derived from thermal dilution cardiac output measurement) and ejection fraction (measured by radionuclide technique) produced a correlation coefficient of 0.92 and a standard error of the estimate of 10.3 mL over a range of 18 to 130 mL (average, 56 mL). The accuracy of volume and ejection fraction measurements with this new ultrasonic method seems comparable to that of other currently used clinical approaches such as radionuclide and angiography. PMID- 2562481 TI - Brachial plexus lesions following cardiac surgery with median sternotomy and cannulation of the internal jugular vein. AB - There are many possible complications after cannulation of the internal jugular vein (IJV) including injury to the brachial plexus. Neurologic injuries can also occur from sternal splitting. The present study looked at the incidence of brachial plexus lesions after cardiac surgery with and without IJV cannulation. Over 12 months, 815 patients were studied after all types of cardiac surgery. In one half of the group, cannulation of the IJV was avoided when possible. Reducing the incidence of IJV catheterization did not lower the overall incidence of brachial plexus lesions (1.8% to 1.4%). However, there was a higher incidence of neurologic lesions in patients with IJV catheters (3.0% to 0.8%) during the entire study period. All 13 plexus lesions were in the C8-T1 distribution, and seven of the patients had a Horner's syndrome on the same side. No posterior first rib fractures could be detected by radiographs. The brachial plexus lesions were transient but the Horner's syndromes were longer-lasting. It is concluded that the injuries are due to compression and traction of the plexus due to stretching and possibly from hematoma formation from the IJV punctures. PMID- 2562482 TI - Discovery and structure-activity relationships of novel alpha-melanocyte stimulating hormone inhibitors. AB - Novel D-amino acid modified, hexapeptide inhibitors of alpha-melanocyte stimulating hormone (Ac-Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Lys-Pro-Val-NH2, alpha-MSH) are described. The discovery of the alpha-MSH inhibitory activity of a known somatotropin (growth hormone) secretagogue, H-His-D-Trp-Ala-Trp-D-Phe-Lys NH2 ([His1, Lys6-]GHRP, I), and its chemical similarity to the alpha-MSH6-11 sequence provided the impetus to investigate the structure-activity relationships of MSH-GHRP hybrid analogues. In this study we compared the melanotropic activity of a series of peptides of the generic formula H-His-Xaa-Yaa-Trp-D-Phe-Lys-NH2 (H [Xaa7, Yaa8, D-Phe10] alpha-MSH6-11-NH2) on the R. pipiens (frog) and A. carolinensis (lizard) skin in vitro bioassays. In summary, D-Phe7-Ala8 substitution (II) in the heptapeptide template yielded an MSH-like agonist of moderately low potency (EC50 ca. 10(-6) M) relative to alpha-MSH; D-Ala7-Ala8 substitution (III) abolished agonist or antagonist activity. alpha-MSH inhibition was effected by MSH-GHRP analogues having D-Trp7-Ala8, D-Arg7-Ala8, D-Trp7-Arg8 or Phe7-Arg8 substitutions. The D-Trp7-Ala8 and Phe7-Arg8 modified derivatives (I and VI) selectively inhibited alpha-MSH on the R. pipiens assay (pA2 = 4.7 and 5.8, respectively), as they did not possess antagonist (or agonist) activities on the A. carolinensis assay. In contrast, the D-Arg7-Ala8 and D-Trp7-Arg8 modified derivatives (IV and V) inhibited alpha-MSH on both the R. pipiens and A. carolinensis assays (pA2 values ranging 5.0-6.0).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562483 TI - Receptor-specific antibodies by immunization with "antisense" peptides? AB - Synthetic peptides whose sequences are specified by RNA complementary to the mRNA coding for peptide hormones have been reported to be useful antigens for the generation of receptor-specific antibodies. We have synthesized an eikositetrapeptide whose sequence corresponds to the complementary strand of the mRNA coding for the sequence of human ACTH(1-24). This "antisense" ACTH(1-24) peptide, "HTCAh," was coupled to bovine serum albumin or thyroglobulin prior to injection into rabbits. The complex proved to be very antigenic, inducing antisera of high titer and specificity. The antisera were tested in ACTH and MSH binding and bioassays, with or without prior purification of IgG molecules. None of the antisera displayed any effect in these assays, nor did they bind to blotted MSH/ACTH receptor protein from Cloudman S91 melanoma cells or to ACTH antibodies. The HTCAh peptide itself did not display measurable association to tritiated or iodinated ACTH(1-24), nor did it displace ACTH(1-24) in a receptor binding assay. However, the peptide bound to a low affinity site of mouse B16 melanoma cells which was independent of the MSH/ACTH binding site and induced melanin formation in these cells, but only at relatively high peptide concentration. Thus, in our hands, the antisense peptide approach using HTCAh as antigen did not lead to receptor-specific antibodies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562484 TI - Angiotensin II receptor binding and actions in NG108-15 cells. AB - NG108-15 cells were grown in culture to confluency and membranes were prepared from the cells to test for the presence of angiotensin II (Ang II) receptors using 125I-sarcosine1, isoleucine8 angiotensin II (125I-SI Ang II). These radioligand binding studies indicated a single class of binding sites with high affinity (KD = 221 +/- 54 pM) that were saturable (Bmax = 27.2 +/- 1.6 fmol/mg protein) and showed characteristic specificity (SI Ang II greater than Ang II greater than Sar1Thr8 Ang II greater than Ang III greater than Ang I greater than des Phe8 Ang II greater than des Asp1, Arg2, Val3 pentapeptide Ang II). To see if these putative receptor binding sites were associated with a functional response, the effect of Ang II on phosphatidylinositide (PtdIns) hydrolysis, expressed as inositol monophosphate (IP1) formation in intact NG108-15 cells preloaded with 3H myoinositol, was determined. Ang II inhibited IP1 formation up to 35% below the basal rate in a dose related manner. The inhibition of PtdIns hydrolysis was prevented by pre-exposure of the cells to SI Ang II, an Ang II receptor antagonist. These results indicate that undifferentiated NG108-15 cells possess Ang II receptors that mediate a reduction in PtdIns hydrolysis. PMID- 2562485 TI - Synthetic amphipathic sequences of surfactant protein-B mimic several physicochemical and in vivo properties of native pulmonary surfactant proteins. AB - A mixture of lipids and proteins unique to the lung lines the airspaces of all mammalian species. This mixture, termed pulmonary surfactant, is essential for normal lung function. We have synthesized selected amino acid sequences of one of the major low molecular weight surfactant proteins to find whether these peptides can duplicate effects of native protein. Peptide/lipid mixtures approximate results found with native surfactant proteins both in vitro and in vivo. Effects found with native proteins or synthetic peptides include association with, and ordering of, surfactant lipid; changes in surface tension with surface compression; rapid adsorption of lipids from subphase to surface; and improvements in oxygenation of surfactant-deficient rats. PMID- 2562486 TI - Total chemical synthesis of ubiquitin using BOP reagent: biochemical and immunochemical properties of the purified synthetic product. AB - The complete ubiquitin molecule (76 residues) has been synthesized by the solid phase method of Merrifield by using BOP as the coupling reagent. The crude product was purified by gel filtration, middle-pressure liquid chromatography and ion-exchange chromatography. Seven mg of a ca. 95% pure peptide was finally obtained; the overall yield of the synthesis was 1%. The final product was controlled by amino acid analysis and sequencing, HPLC and FAB-Mass spectrometry. Synthetic ubiquitin was found to be antigenically active in immunoblotting experiments and in an enzyme-linked immunosorbent assay with anti-ubiquitin antibodies as well as with anti-ubiquitin autoantibodies from autoimmune patients. Its activity was controlled in a conjugation enzymatic system and was found similar to that of commercial ubiquitin. The successful total synthesis of ubiquitin opens the way to the preparation of various stable analogs that should be useful for studying the intracellular metabolism of this molecular and its involvement in the protein degradation pathway. PMID- 2562487 TI - Study of two different enzyme immunoassays for the detection of Mayaro virus antibodies. AB - This paper presents the evaluation of an enzyme immunoassay in which Mayaro virus infected cultured cells are used as antigen (EIA-ICC) and an IgM antibody capture ELISA (MAC-ELISA) for Mayaro serologic diagnosis using 114 human sera obtained during a Mayaro outbreak occurred in Bolivia, in 1987. Results were compared with those obtained by haemagglutination-inhibition test (HAI). MAC-ELISA was the most sensitive technique for anti-Mayaro IgM detection. MAC-ELISA was twice as sensitive as IgM EIA-ICC. The data shows that MAC-ELISA is a practical and valid technique for diagnosis of recent Mayaro infection. IgG EIA-ICC showed high sensitivity and high specificity compared to HAI. The combination of anti-Mayaro IgG and IgM EIA-ICC results presented the highest sensitivity of the study. Anti Mayaro IgG and IgM simultaneous detection by EIA-ICC can be used for recent infection diagnosis (in spite of a less sensitive IgM detection than by MAC ELISA), for surveillance and sero-epidemiologic studies, and for studies of IgG and IgM responses to Mayaro infection. PMID- 2562488 TI - Detection of hepatitis A viral genome in stool samples of patients with relapsed hepatitis A by the polymerase chain reaction. PMID- 2562489 TI - [Renal artery thrombosis in newborn infants undergoing umbilical artery catheterization]. AB - A prospective study to detect renal artery thrombosis by radionuclide renal scintigraphy in newborn infants who underwent umbilical arterial catheterization over a one year period was done: 62 babies were catheterized, 92% were preterm and 85% had severe respiratory distress syndrome that required mechanical ventilation, 25/62 (40.3%) survived and in all of them Tc-99m DMSA scans were taken at a median of 5 days after withdrawal of the umbilical artery catheters (range 9 h to 29 days). A baby with renovascular hypertension had a DMSA scan which showed segmental vascular defect in one kidney. Another patient had left iliac artery thrombosis and two others showed evidence of transient vasospasm. Death occurred in 37/62 (59.7%), 92% of whom underwent autopsy studies which showed aortoiliac thrombosis in 8.8%, all of them without clinical symptoms. Other 12 newborn infants who died without previous umbilical artery catheterization had no evidence of thrombosis at autopsy. PMID- 2562490 TI - Diagnostic role of computed tomography and magnetic resonance imaging. PMID- 2562491 TI - [Model studies for determination of adhesion between prosthetic field and plate prosthesis]. AB - The problem of retention of the complete plate prostheses is complex and determined by a series of factors. The authors follow up the effect of the components of the adhesives for complete prostheses on the alteration of adhesion between the plate prosthesis and mucosa, carrying out model studies. They studied the effect of silicon dioxide and nystatin on the adhesion between the plate prosthesis and mucosa making use of an adhesive based on high molecular neutral polyethylene oxide. A method has been developed for the determination of the effectiveness of adhesives for plate prostheses. PMID- 2562492 TI - Arterial and venous coronary bypass grafts: differential responses to alpha natriuretic polypeptide on the noradrenaline contractures in vitro. PMID- 2562493 TI - Spontaneous defibrillation with blood cardioplegia. PMID- 2562494 TI - Genes, nerve cells, and the remembrance of things past. AB - Learning produces changes in neuronal architecture, changes that result from learned alterations in gene expression. Insofar as psychotherapy resembles learning, it is also likely to produce anatomical changes. These changes should be detectable by imaging methods and ultimately should yield objective evaluations of outcome. PMID- 2562495 TI - The clinical utility of pharmacological agents that act at serotonin receptors. AB - The past decade has seen important advances in the clinical utility of serotonergic agents. The putative novel anxiolytic effects of 5-HT1A partial agonists such as buspirone, the antidepressant effects of selective serotonin (5 HT) uptake blockers such as fluoxetine, and the unique and potent antiemetic effects of 5-HT3 antagonists in cancer chemotherapy are excellent examples of the clinical relevance of selective 5-HT receptor agents. The increasing ability to modulate serotonergic neurotransmission through distinct 5-HT receptor subtypes should greatly facilitate the analysis of 5-HT in both normal and abnormal human brain function. PMID- 2562496 TI - Interactions of paramagnetic contrast agents and the spin echo pulse sequence. PMID- 2562497 TI - In vitro metabolism of 75Se-selenite and 75Se-selenomethionine in chick blood. AB - The metabolism of 75Se-selenite and 75Se-selenomethionine (SEM) in chick blood was studied in vitro. 75Se-selenite was rapidly taken up by erythrocytes and then subsequently released into the plasma in a protein bound form. However, 75Se-SEM showed a more gradual and continuous build up in erythrocytes over the 12 hour incubation period, according to a hyperbolic type function. Chromatography of erythrocyte lysates on Sephadex G-200 indicated that 75Se-selenite was incorporated into glutathione peroxidase, whereas 75Se from SEM was mostly incorporated into hemoglobin. The elution pattern (on Sephadex G-200) of plasma 75Se-selenoproteins from 75Se-selenite or 75Se-SEM showed that radioactivity was associated with two peaks corresponding to molecular weight 440,000 and 89,000. Binding of 75Se from either selenite or SEM to plasma proteins was dependent on the presence of erythrocytes. Addition of reduced glutathione and glutathione reductase to plasma produced the same effects as erythrocytes on binding of 75Se from selenite, but not from SEM, to plasma proteins. Dialysis experiments and treatment with beta-mercaptoethanol before addition of trichloroacetic acid suggested that 75Se in these selenium containing proteins is bound as a selenotrisulfide bond. This was confirmed by chromatography of the released radioactivity on an amino acid analyzer. Based on these results, an in vitro model of selenium metabolism in blood is postulated. PMID- 2562498 TI - Effect of selenite on the toxicity of cis-DDP in mice: estimation of trace elements. AB - A significant accumulation of copper, zinc, iron, rubidium and bromine, in addition to platinum, was observed in kidneys and liver of mice treated with cis DDP. This observation suggests that the toxicity of cis-DDP is due to the overall accumulation of trace elements and not only to the expected high platinum levels. Sodium selenite administration prior to cis-DDP results in fast clearance of all these trace elements from kidneys and liver; control levels are reached within 6 days. This may explain the important reduction in cis-DDP toxicity following selenite administration. An X-ray fluorescence (XRF) facility was used for the evaluation of the trace elements. PMID- 2562499 TI - Studies on the determination of Na-K-ATPase in red blood cell membranes. AB - The present paper is concerned with Na-K-ATPase. In the first part it deals with the effect of such factors as Na+, K+, Mg2+, ATP, pH on activity investigated using a soluble Na-K-ATPase from porcine cerebral cortex. In the second part a method for assaying red blood cell Na-K-ATPase is described. This method consists of specific steps for washing and preparation of red cell membranes, pretreatment of Na-K-ATPase with sodium dodecyl sulfate (SDS) and bovine serum albumin (BSA) and determination of enzymatic released inorganic phosphate. Na-K-ATPase prepared and treated in this way shows a higher activity than the conventionally prepared form of enzyme, as latent enzyme sites are exposed by the SDS treatment. The kinetic behaviour of Na-K-ATPase prepared and assayed by using this method is characterized and compared with that of soluble Na-K-ATPase. PMID- 2562501 TI - The asymmetry [correction of asymetry] of 3H-imipramine binding may predict psychiatric illness. PMID- 2562500 TI - Stable hyperzinchemic condition in patients with Behcet's syndrome. Lack of correlation with in vitro cellular immune response capacity. AB - We have observed a significant increase of zinc concentration in plasma and erythrocytes in a group of Italian Behcet patients, evidencing a rare hyperzinchemic condition associated with the disease. The increased zinc level observed does not show any relation to the presence of therapy or with the impairment of the blastogenic response to HSV1, a specific cellular defect in Behcet disease, confirmed in the present study. The possible biological meaning of these observations is briefly discussed. PMID- 2562502 TI - Bradykinin augments the in vitro migration of nonsensitized lymphocytes. AB - Bradykinin and related peptides have been considered important as mediators of acute inflammation, but their role in the cell-mediated immune response has not been extensively investigated. We have examined the effect of physiological concentrations of these autacoids on the migration of nonsensitized lymphocytes employing an in vitro micropore filter assay system. Bradykinin at a concentration of 0.01-1 nM significantly stimulated the migration of both human peripheral blood and rat splenic lymphocytes. Related naturally-occurring kinins (kallidin, MetLys-bradykinin, desArg9-bradykinin) had a similar effect but other autacoids (angiotensins I and II, histamine, serotonin) were inactive. Bradykinin was more active on freshly harvested lymphocytes than on cells incubated for up to 48 h with or without mitogen. Bradykinin appeared to act predominantly as a chemokinetic agent (augmenting random nondirectional motility), as determined by checkerboard analysis. Bradykinin appeared to exert its effect on lymphocytes predominantly through the B1 class of bradykinin receptors. The migratory response to bradykinin could not be attributed to the release of arachidonic acid metabolites, but stimulated migration could be significantly inhibited by the histamine type 2 receptor antagonist cimetidine. These studies provide a novel mechanism whereby nonsensitized lymphocytes may be recruited to sites of delayed type hypersensitivity reactions. PMID- 2562503 TI - [Ileoanal anastomosis with and without reservoir in children and adolescents with ulcerative rectocolitis and familial polyposis]. AB - We compared the postoperative evolution of patients in whom an ileoanal anastomosis was performed, with (group 1, n:9) an without (group 2, n:7) an ileal reservoir. 3 of the 9 patients in group 1 developed intractable diarrhea, it was necessary to reoperate these children and to construct an ileal reservoir. It was not necessary to reoperate any of the group 2 patients; 2 of whom still have an ileostomy due to the presence of preanastomotic abscess. These 2 last patients where excluded from the following analysis. 2 months after surgery, children in group 1 had 16 +/- 2 bowel movements a day, compared with 8 +/- 2 in group 2, p less than 0.05. Children in group 1 did less well than those in group 2 when the following parameters were compared: fecal incontinence (7/9 vs 0/5), perianal dermatitis (6/9 vs. 1/5), treatment with antidiarrheal agents (5/9 vs 0/5), missing school (4/9 vs 0/5) and diminished social activities (4/9 vs 0/5). We conclude that in children who require colectomy, better results are obtained when an ileoanal anastomosis with ileal reservoir is performed. PMID- 2562504 TI - Uptake and transient expression of chimeric genes in seed-derived embryos. AB - Uptake of DNA in dry and viable embryos of wheat by imbibition in DNA solution was detected by monitoring the transient expression of chimeric genes. Gene expression vectors used in this study contained a neomycin phosphotransferase (NPT) II reporter gene fused to various promoters. Some of the chimeric "neo" genes were shown to yield reproducibly NPT II activity in germinating embryos. This NPT II activity was increased markedly when the neo genes were carried by a vector capable of autonomous replication. Dimers of wheat dwarf virus, a monopartite gemini virus, were thus shown to be effective in amplifying the transient expressed NPT II activity in embryos of several cereals. These and other observations indicate that the observed transient expression really results from DNA uptake and expression in plant embryo cells and is not due to contaminating microorganisms. PMID- 2562505 TI - A detailed study of the periodate oxidation of sialic acids in glycoproteins. AB - Periodate oxidation of terminal N-acetyl- and N-glycoloylneuraminic acid residues in the mucins from edible bird nest substance and pig submandibular gland, respectively, can be carried out under conditions which exclusively give rise to the formation of the C-7 analogues of these sialic acids. In contrast, the C-8 compounds can be obtained in a maximum yield of about 40%. Under identical conditions, N-glycoloylneuraminic acid is oxidized about 1.5 times faster than the N-acetylated derivative. After release of the sialic acids by acid hydrolysis, the characterization of the oxidation products was carried out by TLC, by GLC and GLC-MS of the corresponding pertrimethylsilyl derivatives, and by 500-MHz 1H-NMR spectroscopy. In addition, molar response factors for GLC analysis and extinction coefficients in the orcinol/Fe3+/HCl assay were determined. PMID- 2562506 TI - Specificity towards oligomannoside and hybrid type glycans of the endo-beta-N acetylglucosaminidase B from the basidiomycete Sporotrichum dimorphosporum. AB - We have previously shown that an endo-beta-N-acetylglucosaminidase (EC 3.2.1.96) named "Endo B", isolated from culture filtrates of the basidiomycete Sporotrichum dimorphosporum cleaves asialo-, and to some extent, monosialylated bi-antennary glycans of the N-acetyllactosamine type linked to the asparagine residue of peptide or protein moieties [Bouquelet S, Strecker G, Montreuil J, Spik G (1980) Biochimie 62:43-49]. In the present paper, the substrate specificity of the enzyme towards oligomannoside and hybrid type glycans has been analyzed. The results obtained indicate that ovalbumin glycopeptides containing four to seven mannose residues and bovine lactotransferrin glycopeptides containing four to nine mannose residues were completely hydrolyzed by the enzyme. The degree of cleavage was variable among hybrid type structures, since glycopeptides containing the following glycans: (Gal)1(GlcNAc)3(Man)5(GlcNAc)2; (GlcNAc)3(Man)5(GlcNAc)2;(GlcNAc)3(Man)4(GlcNAc)2 were not hydrolyzed by the enzyme while the percentage of hydrolysis of a glycopeptide containing (GlcNAc)2(Man)5(GlcNAc)2 glycan reached 90%. The bovine lactotransferrin was partially deglycosylated (40%) in the absence of non-ionic detergent while native ovalbumin glycoprotein was not hydrolyzed by the enzyme. The oligomannoside- and the N-acetyllactosamine-type degrading activities present in the culture filtrates were not separated at any step of the purification procedure. Both activities were eluted as a single component with an apparent molecular mass of 89 kDa suggesting that they are located on the same enzyme molecule. Endo B represents a powerful tool for removing oligomannoside- and N-acetyllactosamine type glycans from N-glycopeptides and N-glycoproteins. Moreover, advantages in the use of Endo B in a soluble form as well as in an immobilized form result in its high activity and in its stability to heat denaturation and storage. PMID- 2562507 TI - Conserved sequences in bacterial and viral sialidases. AB - The genes of the bacterial sialidases from Clostridium sordellii G12, C. perfringens A99, Salmonella typhimurium LT-2 and Vibrio cholerae 395 sequenced so far were examined for homologies and were compared with sequences of viral sialidases. Each of the bacterial sialidases contains a short sequence of twelve amino-acids, which is repeated at four positions in the protein. All these sequences exhibit significant similarities. Comparing the repeated sequences of the four sialidases, five amino-acids were found to be highly conserved at defined positions: Ser-X-Asp-X-Gly-X-Thr-Trp. Additionally, most of the distances between the four repeated regions are also conserved among the different sialidases. The conserved bacterial sequences show similarity with sialidases of influenza A H7N1 and H13N9. PMID- 2562508 TI - Stereoselective O-glycosylation of trans-4-hydroxy-L-proline derivatives promoted by silver zeolite. AB - Trans-4-hydroxy-L-proline has been converted to four imino- and carboxyl-blocked derivatives which are suitable for the synthesis of 4-O-glycosyl conjugates. Reaction of these derivatives with 2,3,5-tri-O-benzyl-alpha-L-arabinofuranosyl chloride in the presence of a silver zeolite promoter yielded the blocked beta furanosyl amino-acid conjugates. Deprotection gave trans-4-(beta-L arabinofuranosyloxy)-L-proline which was characterised as its crystalline isopropyl ester. 13C-NMR Data are presented for the compounds described. PMID- 2562509 TI - Lectin binding to the porcine and human ileal receptor of intrinsic factor cobalamin. AB - The purified porcine receptor for the intrinsic factor-cobalamin complex bound to concanavalin A, lentil lectin and wheat germ lectin covalently coupled to Sepharose and was eluted with the corresponding soluble sugars. In contrast, human intrinsic factor bound efficiently to concanavalin A, to some extent to lentil lectin, but only slightly to wheat germ agglutinin. The binding of IF-Cbl to the receptor was inhibited when the receptor was pre-incubated with soluble wheat germ agglutinin, with an inhibition constant estimated to be 1.9 mumol/l. After transfer of the purified receptor from SDS-PAGE to Immobilon, ligand blotting of the purified receptor with iodinated lectin showed that concanavalin A and lentil lectin bound to three (75, 56 and 43 kDa) components but that wheat germ agglutinin bound only to the 75 kDa component. These results showed that the alpha subunit of the receptor could bind to wheat germ agglutinin, resulting in an inhibition of its binding with intrinsic factor. Both binding sites of intrinsic factor and of wheat germ agglutinin could be located near to each other. PMID- 2562510 TI - Different 3' end regions strongly influence the level of gene expression in plant cells. AB - We have investigated the functional role of a 3' end region on the expression of a reporter gene in plant cells. In stably transformed plants, expression of the reporter gene without a plant gene 3' end is variable and depends on the fortuitous presence of polyadenylation signals in the downstream sequences. When the reporter gene is flanked by pBR322 DNA, 3'-processing and polyadenylation occurs at (a) cryptic site(s) within these vector sequences. Using a transient gene expression system, we present a deletion analysis of the 3' end of the octopine synthase gene showing that the most proximal polyadenylation signal per se is not sufficient to ensure expression but that a downstream (G)T-rich sequence is also required. Optimal expression of the fusion gene requires more than 98 base pairs and at most 142 base pairs downstream from the most distal polyadenylation site. We analyzed the expression of chimeric genes with 3' end sequences originating from different plant genes. In the transient expression assay, all constructs direct similar neomycin phosphotransferase II activities. However, in stably transformed tissue, the gene constructs displayed characteristic expression levels which varied as much as 60-fold. This result suggests a role for 3' end sequences in post-transcriptional processes such as efficiency of 3'-processing and/or mRNA stability. PMID- 2562511 TI - Transposable elements can be used to study cell lineages in transgenic plants. AB - The beta-glucuronidase reporter gene has been used to develop a sensitive assay for the excision of transposable elements introduced into transgenic plants. The reporter gene, inactivated by the insertion of the maize transposable element Activator (Ac) into the 5'-untranslated leader, was introduced into the genome of tobacco by Agrobacterium-mediated transformation. Reactivation of the beta glucuronidase gene was detected in transgenic plants using a fluorometric or histochemical assay. Reactivation of the reporter gene was dependent on the presence of the transposase of Ac, and resulted from the excision of the Ac element. This assay, together with the improved methods for visualization, will provide a valuable and rapid method for studying the basic mechanism of transposition in plants and for developing modified transposable element systems suitable for gene tagging in transgenic plants. PMID- 2562512 TI - Cell-autonomous behavior of the rolC gene of Agrobacterium rhizogenes during leaf development: a visual assay for transposon excision in transgenic plants. AB - We describe a genetic switch based on the Ac transposable element of maize and the rolC gene of Agrobacterium rhizogenes, a dominant gene, which has pleiotropic effects on plant growth and morphology. Moreover, rolC gene expression under the control of the 35S cauliflower mosaic virus promoter decreases chlorophyll content in transgenic tobacco plants. Chlorophyll is a visible cell-autonomous marker, and it is shown here that the reduction in chlorophyll content caused by the rolC gene product allows us to monitor, in palisade or spongy mesophyll cells, Ac excision events resulting in rolC gene expression as pale-green sectors and spots. Our results indicate that the rolC gene product behaves in a cell autonomous manner during leaf development, at least as far as chlorophyll accumulation is concerned. In addition, the rolC gene can be useful to evaluate visually if and when a transposable element is active. Most important, we propose the use of a transposable element as a tool to activate expression of morphogenetic genes in a clonal population of cells. This could be particularly useful when studying genes affecting growth and development whose constitutive expression can severely impair regeneration of transgenic plants. PMID- 2562513 TI - The 18-kD protein that binds to the chloroplast DNA replicative origin is an iron sulfur protein related to a subunit of NADH dehydrogenase. AB - From a high-salt extract of the purified thylakoid membrane, an 18-kD protein was detected. This protein was translated by the chloroplast ribosomes and could form a stable DNA-protein complex with a cloned chloroplast DNA replicative origin [Nie, Z.Q., Chang, D.Y., and Wu, M. (1987) Mol. Gen. Genet. 209, 265-269]. In this paper, the 18-kD protein is linked to frxB, a chloroplast-encoded, ferredoxin-type, iron-sulfur protein, by N-terminal microsequencing of the purified protein and computer analysis. The identification is further supported empirically by the fact that the electron paramagnetic resonance spectra of the protein indicate the presence of iron-sulfur clusters. A polyclonal antibody raised against a synthetic pentadecameric peptide with amino acid sequence corresponds to the highly conserved region of the frxB protein and reacts strongly and specifically with the 18-kD protein band in protein gel blot analyses. The 18-kD iron-sulfur protein is found to be related to a subunit of the respiratory chain NADH dehydrogenase by its cross-reaction with a polyclonal antibody raised against highly purified NADH-ubiquinone oxidoreductase, a key enzyme of the respiratory chain. These data are consistent with chlororespiration, and, thus, possible implication of chlororespiration in regulating the initiation of chloroplast DNA replication is discussed. PMID- 2562514 TI - [Cytology and biopsy by fine needle aspiration with ultrasound guidance in abdominal tumors]. AB - To determine the value of fine needle aspiration biopsy (FNAB) we selected 53 patients with liver, pancreas and other abdominal tumors, their diagnosis was made on clinical and ultrasound grounds, we performed FNAB in all these patients. The specimens were sent for cytology and histopathology. 33 out of 53 cases were reported as liver cancer: true positives: 29, false negative 2, and true negatives: 2, for sensibility; 93% specificity: 100%. 15 cases were pancreatic cancer, true positives: 11, true negatives: 3; with 1 false negative, sensibility 91%; specificity 100%. 5 cases were tumors from other abdominal cavity locations (true positives: 3, false negatives: 2; sensibility: 60% specificity: 100%. All these results were well correlated with surgery, clinical follow up and other diagnostic methods ERCP, CT, Laparoscopy. We conclude (FNAB) has a high sensibility and specificity and should be used as a routine procedure in the study of abdominal tumors. No false positive was reported. PMID- 2562515 TI - [Fiber and health]. PMID- 2562516 TI - Epithelial abnormalities in chronic corneal edema: a histopathological study. PMID- 2562517 TI - The influence of refractive error and lattice degeneration on the incidence of retinal detachment. AB - This study indicates the feasibility of stratifying the general population into various risk pools for retinal detachment depending on a person's age, refractive status, and the presence of lattice degeneration. At first impression the risks seem at variance with the fine clinical studies of Byer, who has shown a very low detachment rate in the population with lattice degeneration. In all likelihood the vast majority of his patients were emmetropic or mildly myopic, so that very few would be expected to develop detachments during their entire lifetimes, let along during intervals of only 10 to 20 years. This study shows the futility of following, or treating prophylactically, young emmetropic individuals with lattice degeneration. Assuming that prophylaxis is actually effective, one would have to treat 1000 emmetropic lattice patients in the 30 to 39 year age group to prevent a single detachment over a 10-year period. Lattice patients with low to moderate degrees of myopia tend to develop detachments between 40 and 60 years of age caused by premature posterior vitreous separation and tractional tears. Clearly prophylaxis for this group is not warranted, since only 5% to 10% of these individuals will experience detachments in their lifetimes. On the other hand this study has verified the previous suspicions that persons with myopia exceeding -5.0 D accompanied by lattice degeneration have an extraordinarily high risk of detachment during their lifetimes. Detachments in this group tend to cluster in the second, third, and fourth decades, are typically caused by atrophic holes, are slowly progressive, and are often simultaneously bilateral. Enhanced vigilance is certainly appropriate during this time and perhaps consideration should be given to prophylactically treating this group. This would be no small task, since within a population of 1 million persons there would be about 1150 aged 10 to 39 years with myopia exceeding -5.0 D and lattice degeneration. Only 4 detachments annually and 40 detachments in 10 years would be expected in this highest risk group. PMID- 2562518 TI - Acute accommodative and convergence insufficiency. PMID- 2562519 TI - Botulinum treatment of strabismus in children. PMID- 2562520 TI - Outcome of deteriorated accommodative esotropia. PMID- 2562521 TI - Cytomorphometry of uveal melanomas: fine needle aspiration biopsy versus standard histology. PMID- 2562522 TI - Pyogenic granulomas of the eye and ocular adnexa: a study of 100 cases. PMID- 2562523 TI - Cobalt plaque radiotherapy versus enucleation for posterior uveal melanoma: comparison of survival by prognostic index groups. PMID- 2562524 TI - The tight retracted lower eyelid. PMID- 2562525 TI - Gadolinium enhanced magnetic resonance imaging in the diagnosis of anterior visual pathway meningiomas. PMID- 2562526 TI - Bilateral intracavernous carotid aneurysms presenting as pseudo-ocular myasthenia gravis. PMID- 2562527 TI - The use of a liposome-encapsulated 5-fluoroorotate for glaucoma surgery: I. Animal studies. AB - We have considered the wound-healing and tissue-repair process as it applies to glaucoma filtration surgery. Information derived from classical histologic studies of functioning and failed blebs has been considered in view of the new ideas based on modern cell biology concepts of the wound healing process. These evaluations provide a rationale for the use of adjunctive chemotherapy for glaucoma filtration surgery. A study is described using posterior-lip sclerectomies, an experimental monkey model, a liposomal-drug delivery system and the potent pyrimidine analog L5FO. Results of the clinical evaluation, IOP measurements, and histologic examination of L5FO treated and untreated eyes are reported. We found that the wound healing response at the surgical limbus after a posterior sclerectomy appears similar to that described for other vascularized soft tissues. The macrophage appeared to play an important role in the healing of the episcleral tissues (including Tenon's capsule and the conjunctival stroma), which proceeds rapidly resulting in the deposition of a fibrovascular scar by 2 weeks in most cases. The administration of the L5FO is highly effective in preventing the formation of such scars and thereby promoting development of functioning blebs. The administration of this wound healing retardant 1 week prior to surgery, compared to its administration at the time of surgery, shows that both methods are effective, but the use of L5FO prior to surgery is most effective. The use of the experimental monkey model described by Parrish et al offers many research opportunities to develop better adjunctive chemotherapies for glaucoma filtration surgery. PMID- 2562529 TI - Prospective 5-year postoperative study of cataract extraction and lens implantation. PMID- 2562528 TI - Muller cell alterations from long-term ambient fluorescent light exposure in monkeys: light and electron microscopic, fluorescein and lipofuscin study. PMID- 2562530 TI - Conjunctival provocative tests: a model of human ocular allergy. PMID- 2562531 TI - The surface of the corneal graft: in vivo color specular microscopic study in the human. AB - An in vivo microscopic study of the cellular morphology of the corneal graft surface, employing CSM, has been presented. The following epithelial cellular abnormalities have been noted on the graft surface: a high prevalence (70%) of central vortex keratopathy in the postoperative graft; a redirection of cells in a palisading pattern around sutures; a piling up of cells at the wound junction, with redirection parallel to the wound; cellular evidence of filaments and coarse mucus plaques near the suture line. Vortex keratopathy was not seen in grafts after sutures had been removed, and palisading of cells around sutures disappeared after suture removal. Central epithelial cell morphology in grafts 2 years or more after surgery resembled that of normal patients. A discussion of possible causative factors in the induction of epithelial abnormalities included sutures, corneal denervation, topographical changes, lid pressure, and tension effects on epithelial mitosis. Based on the observations of this CSM study, and based on other laboratory and clinical reports, a new hypothesis concerning epithelial cell movement in the cornea has been presented. It is proposed that there is a differential sliding of epithelium from the periphery to the center, and that epithelium is drawn centripetally by preferential loss of surface cells at the corneal center. Shearing forces of the upper lid cause maximal surface cell loss at the corneal center; shearing forces of the lid are lesser inferiorly and at the limbus. In areas of less lid force, as with depressions near graft sutures, areas of epithelial cell stability are created. Vortex patterns represent an exaggeration of normal cellular pathways of movement, occurring when differential sliding of cells is pronounced, as in corneal grafts. PMID- 2562533 TI - Elevated intraocular pressure following penetrating keratoplasty. AB - IOP was retrospectively studied in 229 consecutive cases of PK, with a mean follow-up period of 84 weeks. Twenty-seven percent of the cases had preoperative glaucoma, three-fourths of which were medically controlled prior to keratoplasty. Following PK, 34% of the total sample developed sustained elevated IOP but only five (2%) patients required surgical treatment for glaucoma. The mean time from PK to first IOP rise was 24 weeks. Variables which were significantly associated with IOP rise included preoperative glaucoma (P less than .001), aphakia (P less than .01), and IOL removal (P less than .01). When eyes with preoperative diagnoses of glaucoma were excluded from the analysis, aphakia was no longer associated with postoperative IOP rise. Keratoconus patients were at significantly less risk than patients with other corneal diagnoses. Factors not associated with IOP rise included previous PKs, pseudophakia, intraoperative vitrectomy, PAS lysis, iridoplasty, secondary IOL placement, and concomitant cataract extraction with IOL (triple procedure). PMID- 2562534 TI - Ultraviolet radiation and the eye: an epidemiologic study. AB - Circumstantial evidence from biochemical, animal, and epidemiologic studies suggests an association between exposure to UV-B radiation (290 nm to 320 nm) and cataract. Such an association had not been proven because it had not been possible to quantify ocular UV-B exposure of individuals or to reliably grade the type and severity of cataract in field studies. We undertook an epidemiologic survey of cataract among 838 watermen who work on the Chesapeake Bay. Their individual ocular UV-B exposure was quantified for each year of life over the age of 16, on the basis of a detailed occupational history combined with laboratory and field measurements of ocular UV-B exposure. Cataracts were graded by both type and severity through clinical and photographic means. SMD changes were ascertained by fundal photography. A general medical history was taken to discover potentially confounding factors. This study showed that people with cortical lens opacities had a 21% higher UV-B exposure at each year of life than people without these opacities. A doubling in lifetime UV-B exposure led to a 60% increase in the risk of cortical cataract, and those with a high annual UV-B exposure increased their risk of cortical cataract over threefold. Corneal changes, namely pterygium and CDK, were also strongly associated with high UV-B exposure. No association was found between nuclear lens opacities or macular degeneration and UV-B exposure. This study also indicated several simple, practical measures, such as wearing spectacles or a hat, that effectively protect the eye from UV-B exposure. Thus it is easily within the power of individuals to protect their eyes from excessive UV-B exposure and reduce their risk of cortical cataract. A program of public education in this area could be a cost-effective means of reducing this important disease. PMID- 2562532 TI - An analysis of corneal endothelial and graft survival in pseudophakic bullous keratopathy. AB - PBK has become an important complication of cataract surgery and a leading indication for keratoplasty. While there are many potential causative factors, erroneous concepts of IOL positioning and design appear to have led to PBK with many iris-supported and anterior chamber lens styles. Underlying host endothelial abnormalities are an important risk factor with posterior chamber lenses. Previous studies of keratoplasty for PBK have shown variable early results in terms of graft clarity and visual rehabilitation. Specular microscopy and life table survival analysis have been infrequently used to study endothelial and graft survival after keratoplasty. This study combined these techniques to evaluate several approaches to the original IOL at PKP for PBK. Four-hundred sixty-nine patients having PKP for PBK between 1976 and 1986 were studied in five retrospective cohorts on the basis of whether their IOL was retained, removed, or exchanged. Specular microscopy was performed prospectively on 390 patients. Survival analysis showed overall failure in 20% of IOL-removed, 24% of IOL retained, and 16% of IOL-exchanged grafts, without significant differences. Within the retained group, however, graft failure rate for posterior chamber IOLs (6%) was significantly less than for anterior chamber (34%) and iris-supported (29%) lenses. With lens exchange, the failure rate was 8% for sutured posterior chamber lenses, 5% for one-piece anterior chamber lenses, and 24% for closed-loop anterior chamber lenses. Graft failure rates exceeded rejection rates for retained iris-supported and anterior chamber lenses, and exchanges for closed loop anterior chamber lenses, suggesting nonimmunologic causes. The survival curve for all groups combined showed cumulative survival of 93% at 1 year, decreasing to 62% by 6 years. Survival was lowest for retained anterior chamber and iris-supported lenses and exchanged closed-loop anterior chamber lenses. Visual acuity results were best for retained posterior chamber IOL eyes and exchange for one-piece anterior chamber IOLs. Exchange for one-piece anterior chamber IOLs gave significantly better visual acuity than exchange for sutured posterior chamber IOLs. There was not a significant relationship between duration of corneal edema prior to PKP and visual outcome, refuting earlier findings. Cystoid macular edema was related to poor vision in 62% of those with visual acuity of less than 20/40 and in 36% of all patients. Specular microscopy findings at 1 year were predictive of longer term survival results. The least cell loss was for retained and exchanged posterior chamber lenses and exchange for one-piece anterior chamber lenses.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2562535 TI - Development of a system for excimer laser corneal surgery. PMID- 2562536 TI - The role of prostaglandins in the para-aminoclonidine-induced reduction of intraocular pressure. PMID- 2562538 TI - Delayed neurotoxicity in monkeys exposed developmentally to methylmercury. AB - Five monkeys (Macaca fascicularis) were dosed from birth to 6.5-7.0 years of age with 50 micrograms/kg/day of mercury as methylmercuric chloride. Blood mercury levels were 0.6-0.9 ppm throughout most of the period of dosing. Blood mercury levels declined rapidly after cessation of dosing to levels below 0.01 ppm. When these monkeys were approximately 13 years old, some individuals displayed clumsiness during routine exercise. This observation was examined by cage-side observation and independent clinical neurological assessment by two staff veterinarians. Fine motor performance was assessed by timing retrieval of raisins from recessed grids. Treated monkeys took longer to retrieve raisins than age matched controls. Clinical neurological assessment revealed apparent insensitivity to touch and pin-prick in mercury-exposed monkeys, and exposed monkeys were clumsier and slower in the exercise cage. These results indicate that overt signs of toxicity can be manifested long after exposure to methylmercury has ceased. PMID- 2562537 TI - Downbeat nystagmus: characteristics and localization of lesions. AB - Clinical examinations and eye movement recordings of 91 consecutive patients with DBN were analyzed to describe the characteristics of DBN and to localize the lesions producing this abnormality. Horizontal and vertical eye movement recordings were made with EOG and/or magnetic search coil. The most frequent causes were infarction, cerebellar and spinocerebellar degeneration syndromes, MS and developmental anomalies affecting the pons and cerebellum. Toxicity from anticonvulsant drugs probably caused nystagmus in a few patients. Clinical examinations, excluding electronic eye movement recordings, were used to localize lesions. Localizations included the cerebellum in 88% of the patients. However, localizations to structures outside of the cerebellum were made in several patients. The effects of DBN of gaze position, convergence, blockage of fixation, and positioning of the head and body were observed. Almost all patients had DBN in some position of gaze while sitting and fixating a distant target. A few patients demonstrated DBN only with convergence, in the dark, or with positioning of the head and body. Horizontal gaze increased DBN in most patients. The nystagmus slow components usually had constant-velocity or increasing-velocity waveforms. The effects of vertical gaze on DBN were variable. In general, statistically significant differences in the frequencies of these effects among the various causes and localizations of lesions were not found. Horizontal eye movements were electronically recorded in DBN patients, in a group of normal subjects, and in a group of patients with isolated cerebellar atrophy who did not have DBN. The pattern of abnormal horizontal eye movements characteristic of damage to the midline structures of the cerebellum (impaired pursuit, impaired OKN, and inability to suppress VOR) was found in almost all DBN patients (99%), including patients with lesions localized to structures outside the cerebellum by clinical examination. DBN is usually produced by lesions in the cerebellum that also damage pathways that control horizontal tracking and visual-vestibulo-ocular interactions. PMID- 2562539 TI - Mercury level and histochemical distribution in a human brain with Minamata disease following a long-term clinical course of twenty-six years. AB - Details are given of the mercury level and deposition in a human brain with Minamata disease following a twenty-six year clinical course after the first severe attack in 1956. This is the first report of a case in which the methylmercury level within the brain has returned to normal limits in a severely affected victim. However, the total mercury remained high in the brain, and mercury was clearly demonstrated histochemically in microglial cells or macrophages over wide areas of the brain and in neurons of specific brain areas. Bergmann's glial cells also contained mercury deposits, whereas Purkinje's cells had relatively little or no deposition. Mild deposition of the metal was demonstrable in the epithelial cells of the choroid plexus. PMID- 2562540 TI - Neurofilament protein crosslinking in gamma-diketone neuropathy: in vitro and in vivo studies using the seaworm myxicola infundibulum. AB - Neurofilament (NF) protein crosslinking has been proposed as the ultimate pathogenetic mechanism underlying the neuropathies caused by the gamma-diketones 2,5-hexanedione (HD) and 3,4-dimethyl-2,5-hexanedione (DMHD). Mammalian models have been used to investigate this hypothesis, but alternative experimental models are needed. Myxicola infundibulum is a marine worm which is gaining popularity in neuroscience research because of its large syncytial axon. A model system using Myxicola has been developed to investigate NF crosslinking in worms exposed to neurotoxic agents whose putative mechanisms involve covalent crosslinking of NF proteins. In vitro studies using purified NF demonstrate that progressive alkylation of Myxicola NF with [2,5-14C]DMHD is accompanied by NF protein crosslinking. Rabbit anti-Myxicola NF antisera showed highly restricted activity for Myxicola axoplasm and NF and were employed for immunoblotting axoplasm from Myxicola treated in vivo with DMHD. A dramatic increase in high molecular weight material was demonstrated in the axoplasm of treated worms, as demonstrated by polyacrylamide gel electrophoresis, and the new high molecular weight bands stained with the anti-NF antisera, indicating the presence of anti NF reactive material in the crosslinked protein. Further, there was progression of crosslinking after cessation of exposure in vivo, an observation which suggests oxidation of remaining pyrrolyl derivatives. These studies support previous observations which suggest that NF crosslinking is the molecular event which initiates NF aggregation in gamma-diketone neurotoxicity, and establish Myxicola infundibulum as a useful species in which to study certain neurotoxic compounds. PMID- 2562541 TI - Members. American Ophthalmological Society. PMID- 2562542 TI - The Tenth Frederick H. Verhoeff lecture. What else did 1864 contribute to ophthalmology? AB - In summary we can say that the AOS was founded at a time when ophthalmology established itself as an independent scientific medical specialty. A hundred years earlier, in 1750, ophthalmology became an independent surgical specialty when Jacques Daviel of Marseille had begun extracting a cataract instead of merely couching or dislocating the lens. Now in the middle of the 19th century a new era dawned on the ophthalmic horizon. An era which Julius Hirschberg calls "the reform of ophthalmology." It was effected mainly by a group of unusual, gifted and genial scholars. Hermann v. Helmholtz, who not only invented the ophthalmoscope, but established with his handbook physiologic optics as an advanced, sophisticated branch of optics and mathematics; F.C. Donders, who put refraction, refractive errors and accommodation on a sound scientific footing, the great A. v. Graefe, who contributed so much to the concept and treatment of glaucoma, to strabismus, to various diseases of the fundus, to neuro ophthalmology and to many other fields and finally William Bowman, the great investigator, clinician and surgeon. It was during this time of reform, of fermentation, of maturation, that a group of farsighted American ophthalmologists decided to establish a society to further the aims and objectives of our specialty in America. The time was right; the effort succeeded and our society developed into one of the decisive forces of American ophthalmology. I hope that my address has met the objectives which I had outlined earlier: To present and illuminate the circumstances and external conditions which were effective in 1864 when our society was founded. At the same time I hope I have done justice to the memory of this outstanding American ophthalmologist, Frederick Verhoeff, who contributed so much to the American Ophthalmological Society. On this the quasquicentennial jubilee of the AOS we find the Society healthy and flourishing. May it continue as an association of the most prominent and most promising American ophthalmologists who consider the practice of our specialty a scholarly profession and not a mercenary trade. I can only conclude with wishing the AOS a happy birthday, many successful returns, ad multos annos! PMID- 2562543 TI - An overview of ocular adnexal lymphoid tumors. AB - In comparison with our earlier colleagues quoted in the introduction, we have made substantial progress in understanding the biology of ocular adnexal lymphoid tumors. While we have refined various categories with prognostic clinical value regarding possible associated systemic disease, none is foolproof and all have varying degrees of unpredictability. Comparatively well-differentiated histologic subtypes predominate among ocular adnexal lymphoid tumors. Polyclonal lesions occur less than half as often as monoclonal B-cell lesions. Molecular genetic studies have revealed small clones of monoclonal populations among the B-cells comprising most of the immunophenotypically polyclonal lesions, but no clonal genetic rearrangements have been uncovered within the preponderant constituent T cell populations. The overall prognosis for ocular adnexal lymphoid tumors is excellent; when lumped together, 67% are not found to be associated with systemic disease with mean follow-ups of over 4 years. This is similar to experience with extranodal and extralymphatic lesions in other sites of the body, which also frequently have a small lymphocyte composition. The incidence of nonocular disease in all categories of our studies, however, will probably increase with the acquisition of longer follow-ups. Careful histopathologic evaluation is as good as immunophenotypic analysis of these lesions in predicting clinical outcome in terms of associated nonocular disease. Polyclonal and well-differentiated B cell monoclonal lesions displayed equivalent clinical behavior. Benign polyclonal lesions may be associated with systemic disease but in a minority of cases (27%), as has also been determined in earlier studies. Clinical staging is the single most important predictor of associated monocular disease. In this study, patients with stage I-E disease had an 87% chance of not developing any nonocular lymphomatous lesion. We believe that this figure may also somewhat decrease with the passage of time. Precise anatomic localization of the lesion within the adnexa had considerable predictive value. Lesions of the conjunctiva fared the best; those of the orbit had an intermediate prognosis; while lid lesions had the worst prognosis. The most favorable prognosis would be held by a conjunctival lymphoid lesion in stage I-E composed of small lymphocytes. The fact that there is a fairly close equivalence in outcome between polyclonal and monoclonal well differentiated lesions indicates that these lesions are in the vast majority of cases primary hyperplasias or primary lymphomas. The discovery by genetic probes of small monoclonal populations in immunophenotypically polyclonal lesions suggests that there is an evolution that goes on in situ.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2562544 TI - Orbital varix thrombosis. AB - Three patients have been described with a thrombosed orbital varix. The clinical, imaging, surgical, and pathologic features of this disorder are described. A pathophysiologic mechanism has been proposed and this entity has been placed in the spectrum of orbital vascular lesions. Recommendations for the diagnostic evaluation and management of patients with thrombosed orbital varices have been offered. PMID- 2562545 TI - The American Ophthalmological Society: the first 125 years. PMID- 2562546 TI - Timolol treatment prevents or delays glaucomatous visual field loss in individuals with ocular hypertension: a five-year, randomized, double-masked, clinical trial. AB - A 5-year, randomized, double-masked, clinical trial was conducted to determine whether treatment with topical timolol maleate was effective in preventing or delaying the onset of glaucomatous visual field loss in subjects with ocular hypertension. Sixty-five individuals considered to be at moderate risk for developing open-angle glaucoma were recruited for the study. In each patient, one eye was chosen randomly to receive timolol twice-daily while the fellow eye received diluent (placebo). Timolol administration reduced IOP from baseline in the treated eyes over the course of the study by a mean +/- SD of 4.9 +/- 3.4 mm Hg. Timolol administration also produced a mean +/- SD contralateral reduction of IOP from baseline in the untreated fellow eyes of 2.9 +/- 3.1 mm Hg. The mean +/- SD difference in IOP between the treated and untreated eyes during the study was 2.3 +/- 2.6 mm Hg. Over the course of the study reproducible visual field loss developed in 4 timolol treated eyes and 10 placebo treated eyes (P = .039, McNemar test). Clinical progressive optic disc cupping was noted in four treated and eight untreated eyes (P = .11, McNemar test). In the 42 subjects who completed a minimum 4-year follow-up, baseline and final optic disc photographs were analyzed using a computer image analysis system to determine changes in the area of disc pallor. The mean +/- SD increase in optic disc pallor was 0.86% +/- 2.4% in the timolol treated eyes and 1.80% +/- 3.6% in the placebo treated eyes. This difference was statistically significant (P = .04, paired t-test). This study provides evidence that medical treatment prevents or delays the onset of glaucomatous visual field loss and optic disc damage in individuals with ocular hypertension. The magnitude of the protective effect of timolol was partially obscured by the contralateral reduction of IOP in the placebo treated fellow eyes. PMID- 2562548 TI - The neurobehavioral consequences of N-methyl-D-aspartate (N-MDA) administration in rats. AB - Evidence is accumulating which suggests that brain damage associated with certain neurodegenerative conditions may be at least partially produced by the overactivation of N-methyl-D-aspartate receptors (N-MDA). To systematically examine the overactivation of N-MDA receptors, N-MDA was administered directly to the frontal cortex and to the hippocampus in rats. It was found that cortical application (1, 2, or 4 mg) had no effect on motor activity 1, 2, or 3 wk after surgery. Four mg N-MDA failed to affect acquisition of a water maze task despite large decreases in cortical width at the site of application. In addition, no alterations in striatal, hippocampal, or cortical cholineacetyltransferase (CHAT) activity were detected after cortical application. Intrahippocampal N-MDA (0, 2.5, 5.0, 10.0, 20.0 micrograms/site) increased motor activity in a dose dependent manner 1, 2, and 3 wk post-surgery. Furthermore, 10 micrograms/site significantly impaired water maze acquisition. Intrahippocampal N-MDA also increased hippocampal CHAT activity and resulted in a loss of pyramidal and dentate granule cells. These results suggest that N-MDA may serve as a useful tool in studying the effects of glutaminergic hyperfunction and its role in neurodegenerative disorders which involve the overactivation of N-MDA receptors. PMID- 2562549 TI - Efficacy of the chelating agent CaEDTA in reversing lead-induced changes in behavior. AB - The chelating agent CaEDTA has been reported to reverse the deficits in intellectual function and performance associated with Pb (lead) exposure in children. However, such studies have not included rigorous controls for the intervention procedures per se. The experiments reported here examined reversibility of performance changes in a rat model based on behavior sensitive to low-level Pb exposure. Rats were exposed to 50 ppm sodium or Pb acetate in drinking water from weaning. Performance maintained under a Fixed-Interval schedule of food reinforcement began at 55 days of age. Following the onset of the characteristic increase in short interresponse times (IRTs) associated with low-level Pb exposure after 35 experimental sessions, Pb treatment was terminated. Animals within both the control and Pb groups were then matched on the basis of performance indices and injected daily for 5 days with either saline, 75 mg/kg or 150 mg/kg CaEDTA. Subsequent changes in F1 performance were monitored for 35-60 sessions. No consistent effects of CaEDTA were detected in control animals. CaEDTA treatment failed to reverse the behavioral effects in Pb exposed animals. If anything, it tended to further increase the proportion of short IRTs. These data suggest that better controlled clinical studies are warranted to evaluate the efficacy of CaEDTA in reversing Pb-induced behavioral effects before its application for these purposes becomes widespread. PMID- 2562547 TI - The role of apraclonidine hydrochloride in laser therapy for glaucoma. PMID- 2562550 TI - Effects of Aroclor 1254 on dopamine and norepinephrine concentrations in pheochromocytoma (PC-12) cells. AB - Pheochromocytoma (PC-12) cells synthesize, store, release and metabolize dopamine (DA) and norepinephrine (NE) in a manner analogous to that observed in the mammalian central nervous system. These cells were used to develop and validate an alternate method to animal testing to assess the effects of a complex environmental mixture of polychlorinated biphenyls (Aroclor 1254) on cellular catecholamine function. Aroclor 1254, at concentrations of 1 to 100 ppm, significantly decreased cellular catecholamine concentrations after 6 hrs. Exposure at 100 ppm for periods of less than an hr increased cellular catecholamine concentrations while longer exposure times (i.e., 1 to 24 hr) decreased cellular catecholamine concentrations. This in vitro depletion of catecholamines is similar to that seen in vivo. Thus, PC-12 cells may be useful for neurochemical evaluation of neurotoxicants with particular reference to effects on catecholaminergic systems. PMID- 2562551 TI - Replication of ciliary vasomotor effects with controlled intravascular corrosion casting. PMID- 2562552 TI - Changes in the zein composition of protein bodies during maize endosperm development. AB - Zeins, the seed storage proteins of maize, are synthesized during endosperm development by membrane-bound polyribosomes and transported into the lumen of the endoplasmic reticulum, where they assemble into protein bodies. To better understand the distribution of the various zeins throughout the endosperm, and within protein bodies, we used immunolocalization techniques with light and electron microscopy to study endosperm tissue at 14 days and 18 days after pollination. Protein bodies increase in size with distance from the aleurone layer of the developing endosperm; this reflects a process of cell maturation. The protein bodies within the subaleurone cell layer are the smallest and contain little or no alpha-zein; beta-zein and gamma-zein are distributed throughout these small protein bodies. The protein bodies in cells farther away from the aleurone layer are progressively larger, and immunostaining for alpha-zein occurs over locules in the central region of these protein bodies. In the interior of the largest protein bodies, the locules of alpha-zein are fused. Concomitant with the appearance of alpha-zein in the central regions of the protein bodies, most of the beta- and gamma-zeins become peripheral. These observations are consistent with a model in which specific zeins interact to assemble the storage proteins into a protein body. PMID- 2562553 TI - Interaction of a developmentally regulated DNA-binding factor with sites flanking two different fruit-ripening genes from tomato. AB - To investigate mechanisms that control fruit development, we have begun experiments to identify proteins that control gene expression during tomato fruit ripening. We focused on the regulation of two different genes, E4 and E8, whose transcription is coordinately activated at the onset of fruit ripening. We report here that a DNA-binding protein specifically reacts with similar sequences flanking the E4 and E8 genes. The E4 binding site is at position -34 to -18 and, therefore, overlaps the region (TATA box) that in many eukaryotic genes serves to determine the efficiency and initiation site of transcription. In contrast, the E8 binding site is distal, located at -936 to -920 relative to the start of E8 gene transcription. Gel electrophoresis mobility retardation experiments indicate that the DNA binding activity that interacts with these two sites increases at the onset of fruit ripening. Taken together, these results suggest that this DNA binding protein may function to coordinate E4 and E8 gene expression during fruit ripening. PMID- 2562554 TI - Molecular biology of the plasma membrane of higher plants. PMID- 2562555 TI - cis-analysis of the wound-inducible promoter wun1 in transgenic tobacco plants and histochemical localization of its expression. AB - The 5' region of the wound-inducible gene wun1, derived from potato, has been sequenced and analyzed for cis-acting elements important in controlling gene expression in transgenic tobacco plants. Different 5' deletion fragments were linked to the reporter gene beta-glucuronidase (GUS) as transcriptional fusions, and the expression of these chimeric genes was analyzed in leaf tissue. Sequences 111 base pairs upstream of the transcriptional start site were not able to drive the GUS expression over background levels, whereas sequences between -111 and 571 showed a slightly higher activity with equal levels of transcription in wounded and nonwounded tissue. The addition of further upstream sequences (-571 to -1022) enhanced the level of expression by a factor between 13 and 370. The expression driven by this fragment was inducible by a factor of twofold to ninefold by wounding. Histochemical analysis of different tissue from transgenic plants that contain wun1-GUS fusions demonstrates wound-inducible and cell specific wun1 promoter activity in plants containing the -1022-base pair fragment. The location of GUS activity appears to be cell-specific, being highest in epidermal cells of leaves and stems and lower in vascular cells. Activity was reduced to levels that could not be detected by histochemical staining in leaves, stems, and roots of plants containing the deleted promoter fragments. Plants that contain the different deletion constructs and plants that carry the -1022-base pair fragment show high expression in anthers and pollen grains that could not be stimulated by wounding. PMID- 2562556 TI - Abscisic acid-responsive sequences from the em gene of wheat. AB - We demonstrate that a chimeric gene containing the beta-glucuronidase (GUS) reporter gene linked to a 646-base pair 5' fragment (-554 to +92) from the abscisic acid (ABA)-regulated Em gene from wheat is correctly expressed in transgenic tobacco. We observe high activity only in embryos of mature seeds, and immature seeds cultured on ABA show enhanced expression. Using a rice transient assay, we identify a 260-base pair fragment (-168 to +92) that accounts for the ABA-specific 15-fold to 20-fold increase in GUS expression. A 50-base pair sequence (-152 to -103) fused 5' in either orientation to a truncated cauliflower mosaic virus promoter (35S) increases GUS activity threefold in the presence of ABA. Insertion of the Em 5'-untranslated region (+6 to +86) between the 35S promoter and the ATG of GUS results in a 10-fold increase in GUS activity in the absence of ABA. These results suggest the following two functional fragments of the Em 5' region: an ABA response element from -152 to -103 and an element between +6 and +86 that quantitatively increases the ABA response. PMID- 2562557 TI - An octopine synthase enhancer element directs tissue-specific expression and binds ASF-1, a factor from tobacco nuclear extracts. AB - We have investigated the expression pattern conferred by a cis-regulatory element (-212 to -154) from the upstream region of the octopine synthase (ocs) gene in transgenic tobacco plants. Analysis of beta-glucuronidase expression driven by the ocs regulatory element revealed a pattern that is tissue-specific and developmentally regulated. In young seedlings, expression is confined primarily to root tips. In older seedlings, expression is stronger and becomes apparent also in the shoot apex. Insertion of a 16-base pair palindromic sequence (-193 to -178), which is included in the regulatory element, into an rbcS promoter results in the expression of rbcS in roots. The 16-base pair palindrome binds activation sequence factor (ASF)-1, a factor from tobacco nuclear extracts that interacts with the sequence between -83 to -63, designated as activation sequence (as)-1, of the cauliflower mosaic virus 35S promoter [Lam et al. (1989). Proc. Natl. Acad. Sci. USA 86, in press]. The in vivo expression patterns and in vitro binding properties of the ocs palindromic sequence are remarkably similar to those of the as-1 element of the cauliflower mosaic virus 35S promoter. These results suggest the involvement of ASF-1 in the transcriptional regulation of the ocs promoter and the 35S promoter. PMID- 2562558 TI - DNA sequences essential for replication of the B genome component of tomato golden mosaic virus. AB - The genome of the geminivirus tomato golden mosaic virus (TGMV) is divided between two DNA components, designated A and B, which differ in sequence except for a 230-nucleotide common region. The A genome component is known to encode viral functions necessary for viral DNA replication, while the B genome component specifies functions necessary for spread of the virus through the infected plant. To identify cis-acting sequences required for viral DNA replication, several mutants were constructed by the introduction of small insertions into TGMV B at selected sites within and just outside the common region. Other mutants had the common region inverted or deleted. All of the mutants were tested for their effects on infectivity and DNA replication in whole plants and leaf discs. Our results indicate that the common region in its correct orientation is required for infectivity and for replication of TGMV B. Furthermore, the conserved hairpin loop sequence located within the TGMV common region and found in all geminiviruses is necessary for DNA replication, and may be part of the viral replication origin. PMID- 2562559 TI - Functional expression of the leftward open reading frames of the A component of tomato golden mosaic virus in transgenic tobacco plants. AB - The genome of the geminivirus tomato golden mosaic virus (TGMV) consists of two circular DNA molecules designated as components A and B. We have constructed Nicotiana benthamiana plants that are transgenic for the three overlapping open reading frames, AL1, AL2, and AL3, from the left side of TGMV A. In the transgenic plants, the AL open reading frames are under the control of the cauliflower mosaic virus (CaMV) 35S promoter. In TGMV infectivity assays, seven of 10 transgenic lines complemented TGMV A variants with mutations in AL1, AL2, or AL3 when co-inoculated with the B component. The 35S-AL construct was transcribed as a single RNA species in the transgenic plants, indicating that AL1, AL2, and AL3 were expressed from a polycistronic mRNA. This differs from the complex transcription pattern in TGMV-infected plants, which contains five AL transcripts. There was no quantitative correlation between the efficiency of complementation in the infectivity assay and the level of expression of transgenic AL RNA in the leaves of a transgenic line. One line that failed to complement defects in AL1, AL2, and AL3 in infectivity assays contained high levels of transgenic AL RNA and functional AL1 protein. These results provide evidence that chromosomal position can affect the cell- and tissue-specific transcription of the 35S promoter in transgenic plants. Comparison of the complementing plants and wild-type infected plants may provide insight into the TGMV infection process and the use of the CaMV 35S promoter for gene expression in transgenic plants. PMID- 2562560 TI - Binding of a pea nuclear protein to promoters of certain photoregulated genes is modulated by phosphorylation. AB - There have been numerous recent reports documenting phosphorylation of DNA binding proteins [Montminy and Bilezikjian (1987); Sorger, Lewis, and Pelham (1987); Hoeffler, Kovelman, and Roeder (1988); Jones et al. (1988); Prywes et al. (1988); Sorger and Pelham (1988); Yamamoto et al. (1988)], and the transcriptional regulatory activity of at least one of these proteins appears to be modulated by this modification [Montminy and Bilezikjian (1987); Yamamoto et al. (1988)]. We report here on a plant nuclear protein, the DNA-binding activity of which is strongly affected by phosphorylation. This protein, AT-1, binds to specific AT-rich elements (the AT-1 box) within promoters of certain nuclear genes encoding the small subunit of ribulose-1,5-bisphosphate carboxylase and the polypeptide components of the light-harvesting chlorophyll a/b protein complex. A consensus sequence of AATATTTTTATT was derived for the AT-1 box. We demonstrate that the DNA-binding ability of AT-1, from nuclear extracts of pea, can be reversibly modulated by phosphorylation. AT-1 is active in the nonphosphorylated form and loses all DNA-binding ability as a result of phosphorylation. The kinase that phosphorylates AT-1 uses both Mg-ATP and Mg-GTP as a substrate and is inhibited by heparin and spermine, indicative of an NII-type casein kinase. PMID- 2562561 TI - Kunitz trypsin inhibitor genes are differentially expressed during the soybean life cycle and in transformed tobacco plants. AB - We investigated the structure, organization, and developmental regulation of soybean Kunitz trypsin inhibitor genes. The Kunitz trypsin inhibitor gene family contains at least 10 members, many of which are closely linked in tandem pairs. Three Kunitz trypsin inhibitor genes, designated as KTi1, KTi2, and KTi3, do not contain intervening sequences, and are expressed during embryogenesis and in the mature plant. The KTi1 and KTi2 genes have nearly identical nucleotide sequences, are expressed at different levels during embryogenesis, are represented in leaf, root, and stem mRNAs, and probably do not encode proteins with trypsin inhibitor activity. By contrast, the KTi3 gene has diverged 20% from the KTi1 and KTi2 genes, and encodes the prominent Kunitz trypsin inhibitor found in soybean seeds. The KTi3 gene has the highest expression level during embryogenesis, and is also represented in leaf mRNA. All three Kunitz trypsin inhibitor genes are regulated correctly in transformed tobacco plants. Our results suggest that Kunitz trypsin inhibitor genes contain different combinations of cis-control elements that program distinct qualitative and quantitative expression patterns during the soybean life cycle. PMID- 2562562 TI - Soybean beta-conglycinin genes are clustered in several DNA regions and are regulated by transcriptional and posttranscriptional processes. AB - We investigated the chromosomal organization and developmental regulation of soybean beta-conglycinin genes. The beta-conglycinin gene family contains at least 15 members divided into two major groups encoding 2.5-kilobase and 1.7 kilobase embryo mRNAs. beta-Conglycinin genes are clustered in several DNA regions and are highly homologous along their entire lengths. The two groups differ by the presence or absence of specific DNA segments. These DNA segments account for the size differences in beta-conglycinin mRNAs. The 2.5-kilobase and 1.7-kilobase beta-conglycinin mRNAs accumulate and decay at different times during embryogenesis. By contrast, genes encoding these mRNAs are transcriptionally activated and repressed at the same time periods. Our studies indicate that the beta-conglycinin family evolved by both duplication and insertion/deletion events, and that beta-conglycinin gene expression is regulated at both the transcriptional and posttranscriptional levels. PMID- 2562563 TI - A frameshift mutation prevents Kunitz trypsin inhibitor mRNA accumulation in soybean embryos. AB - We investigated the molecular basis of a soybean Kunitz trypsin inhibitor (KTi) gene mutation that prevents the accumulation of Kunitz trypsin inhibitor protein during seed development. We found that mRNA encoding the major Kunitz trypsin inhibitor protein (KTi3 mRNA) is reduced at least 100-fold in null (KTi-) embryos but that KTi3 gene transcriptional activity is similar in Kunitz trypsin inhibitor producing embryos (KTi+) and in KTi- embryos. We sequenced the Kunitz trypsin inhibitor KTi3 gene from both KTi3+ and KTi3- lines and found that these genes differ by only three nucleotides (+481, +486, and +487) within the KTi3 coding region. Alteration of these nucleotides results in a frameshift within the KTi3- gene that causes premature termination during translation. Our results suggest that the KTi3- frameshift mutation results in KTi3- mRNA destabilization and leads to a drastic reduction in KTi3 mRNA prevalence. PMID- 2562564 TI - Tissue-dependent plastid RNA splicing in maize: transcripts from four plastid genes are predominantly unspliced in leaf meristems and roots. AB - Most plastid gene products do not accumulate to high levels in meristem proplastids or in the specialized plastids of roots. To assess whether a modulation of plastid splicing activities might play a role in this tissue dependent expression of the plastid genome, the ratio of spliced to unspliced transcripts from the atpF, petB, petD, and rpl16 genes was compared between several tissues of maize. Although these transcripts are predominantly spliced in green leaf tissue (both bundle sheath and mesophyll cells), spliced atpF, petB, and petD transcripts are underrepresented relative to their unspliced precursors in roots and leaf meristems. The ratio of spliced to unspliced rpl16 transcripts varies in a similar fashion, but the magnitude of the differences between tissues is not as great. The proportion of RNA that is spliced reflects the tissue of origin and not photosynthetic competency, chlorophyll content, or exposure to light since the leaves of photosynthetic mutants and of seedlings grown in the absence of light contain spliced and unspliced transcripts in normal ratios. These results raise the possibility that low RNA splicing activities are in part responsible for the limited expression of the plastid genome in meristematic and root tissue. PMID- 2562565 TI - Role of posttranslational cleavage in glycinin assembly. AB - Glycinin, like other 11S seed storage proteins, undergoes a complex series of posttranslational events between the time proglycinin precursors are synthesized in endoplasmic reticulum and the mature glycinin subunits are deposited in vacuolar protein bodies. According to the current understanding of this process, proglycinin subunits aggregate into trimers in endoplasmic reticulum, and then the trimers move to the vacuolar protein bodies where a protease cleaves them into acidic and basic polypeptide chains. Stable glycinin hexamers, rather than trimers, are isolated from mature seeds. We used a re-assembly assay in this study to demonstrate that proteolytic cleavage of the proglycinin subunits is required for in vitro assembly of glycinin oligomers beyond the trimer stage. The possibility that the cleavage is a regulatory step and that it triggers the deposition of 11S seed storage proteins as insoluble aggregates in vivo is considered. PMID- 2562566 TI - [Fluorine content in natural cow milk from the region of Plovdiv]. AB - Fluorine content in natural cow milk has been determined with a view to the fluoration of the milk and its use for dental caries prophylaxis. Fluorine has been determined by the method of standard calibration, making use of fluorine selective electrode and "Orion SA 720 pH/ISE Meter" with LCD display. The mean fluorine quantity in milk is mean = 0.046 ppm, being far lower than the optimal fluorine concentration. PMID- 2562567 TI - [Comparative study of adhesive properties of some adhesives for complete prostheses]. AB - The adhesives for prostheses are intended to improve the retention of that kind of constructions and to increase the masticatory ability of the patient. The authors studied comparatively the adhesive properties of some of the often used adhesives for complete prostheses--Kukident, Super Poly Grip, Co Re Ga, Badident and Badident N. The highest values of unstuck strength have Badident and Badident N--86.30 and 77.48 MN/m2.10(-4) respectively. PMID- 2562568 TI - [Inquiry data for stomatological health education of pregnant women in the town of Sumen and the region]. AB - The purpose of the authors with the present study was to obtain data on the information of the pregnant women about problems associated with the stomatological health. Anonymous inquiry was carried out among 250 pregnant women from the town of Sumen and the region. The questionnaire has 7 questions. The answers give information on the knowledge about problems associated with nutrition, fluorine prophylaxis, formation of dental germs and cutting. The results reveal insufficient knowledge in all problems. Conclusions are drawn that the micro-courses during the last months of pregnancy in stomatological prophylaxis are ineffective. The individual work proved to be insufficient as well as the participation of the regional and workshop stomatologists. PMID- 2562569 TI - [The rational coding of stomatological data]. AB - The introduction of the computer technique in the stomatological science and practice actualized the problems of coding of stomatological information. The material presented discusses the problems of devising of optimal code system, intended for the stomatological symptomatics. An original mode for semipontional coding is proposed. Basic characteristics of the code system are described, stressing up its main advantages. PMID- 2562570 TI - [Diagnostic difficulties in one patient with chronic periostitis of the lower jaw]. AB - The problem of chronic periostitis of the jaws and the difficulties that sometimes arise in differentiation from a tumourous process is briefly discussed. A case of a ten year old child with chronic ossifying periostitis is described that caused considerable differentiation-diagnostic difficulties, mainly because of the X-ray findings of orthopantomography and computer tomography, presenting data suggesting more a tumourous disease of the lower jaw. After the extraction of the sixth tooth of the lower jaw, with a chronic periapical process, and after antibiotic treatment, the disease was healed. PMID- 2562571 TI - [A child with congenital skin fistula of the sublingual salivary gland]. AB - A patient is described, treated at the clinic for a disease, being a casuistic rarity. A child of 6 is concerned with a congenital skin fistula in the region of the neck, arising from the sublingual salivary gland. The treatment covers the radical extirpation of the gland with excision of the fistula duct. After an uneventful postoperative period, complete healing was attained. PMID- 2562572 TI - [Some data about medicine and dental treatment throughout our lands till the foundation of the first Bulgarian state]. PMID- 2562573 TI - [Dental service in the town of Kazanlak and the region in the past]. PMID- 2562574 TI - [Thermometric diagnostics of chronic ulcerous pulpitis]. AB - Thermometric measurements were performed of 408 teeth of the upper and lower jaw with clinically confirmed initial and advanced stage of chronic ulcerous pulpitis in 398 subjects, aged from 18 to 30. Temperature elevation within the range from 1.4 to 3.2 degrees C was established in the initial form of chronic pulpitis and from 1 to 2.5 degrees C in advanced form of chronic pulpitis as compared with the norm. Significant difference in the temperature deviations exists in both forms of chronic pulpitis between the teeth of the upper and lower jaw. PMID- 2562575 TI - [The role of tartar in the light of the modern clinical-epidemiological, morphological and immunological studies]. AB - After the public recognition of the dental plaque as the primary etiological factor for the diseases of gingiva and periodontium, the tartar is the object of comparatively small number of studies. The author discusses the publications during the last 25 years, grouping the results from the epidemiological, clinical, morphological and immunological studies. Throwing light on new aspects of the pathogenetic role of the tartar in maintaining and chronification of the gingival inflammation as well as in osseous resorption, puts new accent on the necessity of its removal in prophylaxis and therapy. The importance of tartar as a reservoir of antigen substances associated with the total reactivity of macro organisms is stressed upon. PMID- 2562576 TI - [Some new points of view of cavity form]. AB - The discovery of new data about the anatomy and physiology of the dental tissues, the improvement of the diagnostics of dental caries and the development of stomatological materials and technique are preconditions for a change of some concepts for the preparation of the cavities and their form. A new trend in cavity preparation is discussed on the base of literature data, characterized by maximum sparing of the hard dental tissues and a change in some of the geometric characteristics of the cavity form but the basic requirements for the biological and technical expedience remain unaltered. PMID- 2562577 TI - [Application of chlorhexidine in stomatological practice--cons and pros]. PMID- 2562578 TI - [Synthesis of eumelanin pigment precursors. II. 6-hydroxy-5-methoxy- and 5 hydroxy-6- methoxyindole]. AB - Following up of specific melanogenesis metabolite excretion in the course of malignant melanoma disease is useful for the disease prognosis assessment. The authors elaborated a modified synthesis technique of 6-hydroxy-5-methoxy and 5 hydroxy-6-methoxyindole, eumelanin pigment precursors. The elaborated synthesis is economically and temporally reasonable and the synthesized compounds accord, as proved by means of elementary analysis, thin layer (TLC) and high performance liquid chromatography (HPLC) and last but not least by nuclear magnetic resonance (NMR), with demands for reference compounds needed for isomer hydroxymethoxyindole quantification in urine. PMID- 2562580 TI - [Experimental substitution of carbimazole for Alkiron in the atherogenic diet]. AB - Due to present unavailability of Alkiron the authors tried to substitute it by Carbimazole in atherogenic diet aimed at inducing advanced atherosclerosis in rats. Carbimazole proved to be entirely ineffective in this respect and thus the thiouracil derivatives cannot be substituted by it. PMID- 2562579 TI - [The effect of Listeria factor Ei, BCG vaccine and a combination of both on the growth of experimental syngeneic tumors in inbred mice]. AB - The investigation assessed, whether a nonspecific single and repeated immunization induced by Listeria monocytogenes Ei factor, BCG vaccine and their combination influences the growth of transplanted syngenic methylcholanthrene tumour depending on tumor graft cellularity. The cellularity of 10(4) and 10(5) were investigated. Tumour growth was examined 10 and 17 days after the inoculation. It has been found that single Ei factor administration did not influence the tumour growth. Single BCG vaccine favourably influenced the tumour growth, but in progressing time and cellularity the effect was failing. In combination of BCG vaccine and Listeria Ei factor the antitumour effect appeared to be most profound. In progressing time after the tumour grafting and graft cellularity the effect was disappearing. It was probably due to the fact that immunotherapy fails to control tumour mass with high cellularity. The antitumour effect potentiation by combination of BCG vaccine with Ei factor suggests a way to a possible combined antitumour immunotherapy. PMID- 2562582 TI - [Methods of diagnostic bone biopsy and its complications]. AB - By the end of 1986 559 assessable biopsies had been taken at 3rd internal department. The authors continue the previous study performed at the department and suggest further methods for bioptic sampling enabling sampling of larger bone cylinders. They suggest several ways of bone trepan composition adjustment. The authors discuss and compare the incidence of complications associated with bone biopsy performance. They conclude that in inpatients bone biopsy performed in short-term general anaesthesia is a safe intervention. PMID- 2562583 TI - [Additional observations on age differences in the blood circulation in rats]. AB - By means of microparticles labelled by radioactive strontium 85Sr (3M, USA) the authors in two experiments established cardiac output and local blood flow through bones and certain other organs and tissues of rats in two distinct age categories (35 and 90 days, 45 and 110 days). The results of both experiments showed 1) significant decrease of blood flow in skull, lumbar vertebrae, distal epiphysis, distal metaphysis and thighbone diaphysis and tibia in older rats of both sexes (proof of previous results with 86Rb), 2) constantly significant decrease of blood flow through a skin sample taken from the ventral side of the trunk and 3) nonconstant increase of blood flow through kidneys of older rats. The causes of observed changes are not known, physiological importance to be supposed particularly as regards the circulatory changes in bones, is also not clear. PMID- 2562581 TI - [Reactivity of LK-1 monoclonal antibodies with human hematopoietic cells]. AB - A murine monoclonal antibody LK-1 reacting with the common leukocytic antigen gp 95 was prepared by means of standard hybrid technology. This antigen was found in wider distribution on various morphologic types of human blood cells of the monocytic, granulocytic, thrombocytic, erythroid and lymphoid series (with the exception of some B lymphocytes). Furthermore, the monoclonal antibody reacted with the antigen occurring on leukaemic cells of patients with AML, CML, AMoL, ALL and AMoL and reacted with cells of some human cell lines as well. PMID- 2562584 TI - [Evaluation of automatic sphygmomanometers]. AB - In clinical evaluation of a new type of automatic sphygmomanometer, a strict comparability of results with the standard indirect auscultatory method is required. We have compared the oscillometric apparatus BP 103 N Nippon Colin (B) with the Hawksley random zero mercury sphygmomanometer (A) in a stratified sample of 26 subjects using a cross-over design of four simultaneous pairs of measurements in both arms by two observers. Mean values obtained by machine B were higher than values obtained by machine A, by 0.56 kPa systolic and by 0.62 kPa diastolic. Analysis of variance showed that the differences between both machines were within the random variability of blood pressure values obtained, and therefore irrelevant at single measurements. In case of blood pressure measurements in an epidemiological study, a correction of values should be done using the regression model B = K. Ac. PMID- 2562585 TI - [Oxytocinase in pregnancy complications]. AB - Using a simpler method for cystylaminopeptidase (oxytocinase) investigation modified by means of p-nitranilide the oxytocinase activity was examined in blood serum of 51 pregnant women with complicated pregnancy in the period of week 31-34 of gestation. Lower oxytocinase values were found in women with complicated course of pregnancy compared to those in women with physiological course of pregnancy and the same gestational age. Significantly low oxytocinase activity values were established in pregnant women with preterm uterine activity. A significant variability was found in the whole group of 51 women with complicated pregnancy as well as in individual groups. PMID- 2562586 TI - [Difficulties in the use of the Ridley and Jopling classification--a morphological analysis]. AB - There are many difficulties in the use of the Ridley & Jopling classification in daily practice. The author identified the morphologic parameters whose variations permit to distinguish the polar types and borderline groups according to Ridley. If we avoid the inconstant histologic alterations we believe that this distinction depends basically on the following parameters: epithelioid cell, granuloma of epithelioid cells, numbers of lymphocytes and number of bacilli. A critical analysis is performed of each of these parameters and the author concludes that they are scarce, and that there are great difficulties for the identification and interpretation of their variations for classification purposes. These difficulties are even more important during the reactional episodes. PMID- 2562587 TI - [Efficacy of multidrug therapy in treatment of paucibacillary leprosy patients- preliminary results]. AB - The efficaciousness evaluation of the polychemotherapics regimens recommended by WHO to the hansenian paucibacillary patients, is carried out mainly by the suitable follow up of patients after therapeutic discharge. The criterion for inclusion of patients as paucibacillary ones is another point of importance. The authors based on the follow up of 66 patients that completed treatment and in the absence of relapses until the moment, advise that together with the clinical classification it should be considered factors like: the Mitsuda test, the number of lesions and the bacilloscopy result. PMID- 2562588 TI - [Prevention and treatment of physical disabilities in Hansen's disease in Brazil: study of specific professional risk for the disease]. AB - As a continuation of the research line related to the insertion of prevention and treatment of physical disability in programs of leprosy control, we present here results related to specific professional risk for the disease based on data obtained in a recent multicenter investigation at the national level in Brazil. In a survey of 416 professionals and 69 health units, 179 patients with different clinical forms of leprosy were surveyed. Using initial and current patient socioprofessional condition (whether economically active or dependent and the different modalities of the two situations) as an indicator of social mobility, the associations were tested using the chi 2 statistical parameter of Pearson. The results revealed statistical significance at the 5% level among the variables investigated both in the first and the second specific determination. PMID- 2562589 TI - [Myocardial disease, heart failure and reactional tuberculoid leprosy]. AB - A 60-year-old woman was admitted with congestive heart failure, essential hypertension and abdominal distension. Her son reported that she appeared with red spots in the body and that she was under dapsone therapy. Seven months ago there was sudden increase of the skin lesions. In the 11th day after admission she underwent a stroke that progressed to decerebration and she expired on the fourth day. Autopsy confirmed CHF due to chronic myocarditis related to Chagas' disease. Aneurysm of the apical region of the left ventricular chamber was also observed leading to thrombosis and systemic embolism with brain and spleen hemorrhagic infarct. In the encephalous there was edema, uncus herniation and hemorrhagic infarct of the brain stem. The skin lesions were due to reactional tuberculoid hanseniasis (RHT) with focal lesions in axillary lymphnodes, nasopharyngeal mucosa and in the posterior tibial nerve. The pathogenesis of RHT is discussed as well as its differentiation with the BT group of Ridley and Jopling and its probably relationship with the secondary tuberculoid hanseniasis reported by Ridley. The focal lesions are also discussed with END to the involvement of a peripheral nerve trunk what is said to be uncommon in this form of Hansen's disease. PMID- 2562590 TI - Histogenic differentiation of polar tuberculoid granuloma and the "benign" or "malignant" behaviour of hanseniasis. AB - Based on a new morphological concept and classification of granulomatous inflammation: the polar granulomas, on the histogenesis of the tuberculoid granuloma of the positive Mitsuda test and on the relationship between the degree of histogenetic differentiation and behaviour of tumors, the Authors concluded that the "benign" or "malignant" behaviour of hanseniasis depends on the degree of tuberculoid differentiation of the lesions. If the lesion is histologically well differentiated toward a polar tuberculoid granuloma (tuberculoid hanseniasis) it will have a "benign" behaviour. On the contrary, if this differentiation is absent (virchowian hanseniasis) or poor (interpolar borderline hanseniasis) the behaviour of the lesion will be "malignant". PMID- 2562591 TI - [Hansen's disease in the northern region of Brazil--1986]. AB - Some general facts about Hansen's Disease are presented: in the world, in the Americas, in 1986, with the purpose to focalize on the Northern Region of Brazil (Amazon Valley) where an attempt is made to specify, State, the clinical forms in the active registry, the respective coefficients of Prevalence, about the cases detected during the year, by clinical forms and the correspondent Coefficient of Incidence, the age range of less than 15 years, and above 15 years; and to analyze according to the township in the State of Amazonas in a series of 8 years, from 1979 to 1986, the registered cases under control and also without control, the new cases, by clinical form and Coefficient of Incidence, the positive cases among students in the capital and also in the country, and with more details, still by township and by Public Health Registry, in 1986, with the population, new cases, and Coefficient of Incidence, cases of the active registry, cases under control, Coefficient of Incidence, the relationship between patients/inhabitants, cases according to the decreasing intensity as far as the Prevalence and Health Department Regions: all this to give an idea, in detail, inclusive statistics, of the real position of Hansen's Disease in the Northern Region (Amazon Valley), with the highest figures of Prevalence and even of Incidence, in comparison with other Regions of the country, and to stress the grave situation of the endemic proportions of the disease, in Brazil in general and in that Regions, in particular. PMID- 2562592 TI - Teenage pregnancy in western Sydney. AB - The obstetric performance of 121 young teenagers (less than 17 years of age) confining as public patients at Blacktown District Hospital was retrospectively compared with a randomly-selected control group of older gravidas. These adolescents were less likely to be married, to be certain of their last menstrual period, to book into the hospital early or to attend the antenatal clinic regularly than their older counterparts. However, there was no difference in the rates of anaemia, spontaneous or operative delivery, gestational age at confinement, birth-weight, perinatal mortality or neonatal morbidity rates. When the teenagers were compared with primigravidas from the control group, labour length was significantly shorter and the rates of preeclampsia were similar. These results are more optimistic than previous overseas reports regarding the obstetric performance of young adolescents. PMID- 2562593 TI - Trial of labour after previous caesarean section: obstetric outcome. AB - Of 305 patients with a previous lower segment Caesarean section scar admitted over a 28-month period, 207 were allowed a trial of labour. A successful trial of labour was achieved in 63.3% of patients with a recurrent indication and 73.4% with a nonrecurrent indication. Of 75 patients who received oxytocin for augmentation and 22 for induction of labour, 70.5% achieved vaginal delivery. This was similar to the vaginal delivery rate in patients who did not require augmentation in induction. Three cases of scar dehiscence occurred in patients who had oxytocin, but in whom the recommended management protocol was ignored. The events that led to these 3 dehiscences is described. Analysis of birth weights revealed a trend towards more repeat Caesareans with increasing birth weight beyond 2,500 g. This was especially reflected by the higher emergency Caesarean section rate in those who had a trial of labour. A trial of labour in patients with a previous Caesarean scar is safe, and can be allowed even in patients who had the previous Caesarean for cephalopelvic disproportion, although malpresentation and obvious disproportion must be excluded. Judicious use of oxytocin for a limited period of time should help in reducing the number of repeat Caesarean sections. PMID- 2562594 TI - Changes in smoking and drinking during pregnancy. AB - Changes in alcohol and nicotine usage during pregnancy are reported from a sample of 40 pregnant women, 28 of whom smoked prior to pregnancy and 32 of whom drank alcohol. Although most women wanted to reduce or stop their intake during pregnancy, the drinkers were far more successful in their attempt than the smokers. Thus, 85% of drinkers achieved or bettered their target behaviour (in terms of consumption), while 53% of smokers 'failed' in their attempt. Women can identify reasons which influence their smoking and drinking behaviour during pregnancy and, in particular, the amount of emotional attachment to the unborn child appears to be important. However, endorsement of a particular reason does not appear to relate in any simple way to actual consumption, and a major disparity between health beliefs and actual behaviour was apparent. Implications for primary prevention programmes and future research are discussed. PMID- 2562595 TI - Internal iliac and ovarian artery ligation in the control of pelvic haemorrhage. AB - Bilateral ligation of the internal iliac artery with or without ligation of the ovarian arteries was carried out in 17 patients who had life-threatening pelvic haemorrhage; 14 were due to obstetric haemorrhage, 2 followed abdominal hysterectomy and 1 intractable haemorrhage associated with carcinoma of the cervix. The procedure was successful in arresting the haemorrhage in 13 patients but 4 needed hysterectomy in spite of the ligation; 1 patient died from disseminated intravascular coagulation following surgery and 1 had cardiac arrest due to extensive blood loss during surgery and died 12 hours later. A review of the literature, a brief description of the procedure and its indications are discussed. PMID- 2562597 TI - Insulin-like growth factor 1 as an intra-ovarian hormone--an integrated hypothesis and review. AB - Fourteen ovulatory patients undergoing diagnostic laparoscopy had at least 2 samples of clear follicular fluid (FF) collected in the late follicular phase. Dominant follicles contained significantly higher concentrations of insulin-like growth factor-1 (IGF1), oestradiol (E2), progesterone (P) and lower androstenedione (AD) than their matched cohorts. After the luteinizing hormone (LH) surge, a significant increase in dominant FF-IGF1 and FF-P occurred. Log FF E2 and log FF-P were correlated with both FF-IGF1 and FF volume. Log FF-AD was negatively correlated with FF-volume, FF-E2 and FF-P (but not FF-IGF1). The cohort concentration of FF-IGF1 was correlated with serum IGF1. These results support the hypothesis that IGF1 has a central role in the selection of the dominant follicle. PMID- 2562598 TI - Langerhans cell population in early invasive squamous cell carcinoma of the uterine cervix. AB - Langerhans cell counts were performed using indirect immunoperoxidase staining for S100 protein in 10 cases of early invasive squamous cell carcinoma of the cervix and 20 controls from routine hysterectomy specimens. There was a significant (p < 0.01) reduction in the number of Langerhans cells per unit sectional area of tumour tissue compared to the normal cervical epithelium. The density of Langerhans cells in the 3 patients with tumour metastasis in pelvic lymph nodes was further reduced. The low density of Langerhans cells in invasive squamous cell carcinoma compared to the normal state may reflect poor local immunosurveillance and may be related to progression and spread of the tumour. PMID- 2562599 TI - Evaluation of serial cervical cytology in the assessment of preinvasive cervical neoplasia. AB - Examination was made of the cervical cytology in 107 patients who underwent cone biopsy with a subsequent diagnosis of severe dysplasia or carcinoma in situ (105) or microinvasive carcinoma (2). Multiple smears had been performed for each patient over a period of time. From the total number of smears a 'false-negative' cervical cytology rate of 10.3 per cent was found. The time interval from the initial abnormal smear until definitive surgical treatment was carried out was calculated. The cervical smear prior to cone biopsy was found to correlate with the histological diagnosis in only 46.7 per cent. The implications of performing repeated cervical cytology are discussed and a firm recommendation is made that patients with abnormal cervical smear be promptly referred to definitive treatment. PMID- 2562603 TI - Place of second-look laparotomy after 18 courses of chemotherapy in epithelial ovarian cancer. AB - Thirty-two patients with epithelial carcinoma of the ovary underwent a second look laparotomy after the completion of an average of 18 courses of adjuvant chemotherapy. Nine patients (28%) were found to have persistent disease. This low rate of positive second-look laparotomy is probably because these patients received chemotherapy for 18 months and in those patients whose tumour did not respond to the medication, the disease would progress and the patient died without becoming a candidate for laparotomy. There was no stage Ic patient whose second-look laparotomy was positive and the need for second-look laparotomy is questionable in such patients. Stage III/IV patients and patients with recurrence had a significantly higher positive second-look laparotomy rate than stage Ic/II patients, 44% and 7% respectively. In the present group of patients with positive tumour at second-look laparotomy, 33% responded to further debulking operation and chemotherapy and remained disease free 15-33 months after completion of their second line chemotherapy treatment. For second-look laparotomy to be of maximum benefit to the patient, we recommend that it be performed after 12 courses of chemotherapy. PMID- 2562605 TI - Transdermal oestrogen for postmenopausal women: a double blind crossover comparative study with ethinyl oestradiol. AB - A double blind crossover study of transdermal oestradiol (50 micrograms/day) and ethinyl oestradiol (20 micrograms/day) was conducted with 25 postmenopausal women. Transdermal oestradiol increased circulating levels of oestradiol and oestrone. Both preparations favourably improved patients' symptoms and vaginal cytology, lowered gonadotrophin levels and urinary calcium loss and gave a satisfactory menstrual pattern. Transdermal oestradiol had no effect on measures of hepatic function whereas oral ethinyl oestradiol significantly altered levels of sex hormone binding globulin, plasma renin substrate and lipoproteins. Transdermal oestradiol has a comparable beneficial effect on postmenopausal symptoms to ethinyl oestradiol without the adverse effects on hepatic proteins. PMID- 2562608 TI - Vaginal dystocia in a patient with a double uterus and a longitudinal vaginal septum. AB - A case of a breech straddling a vaginal septum requiring urgent division is reported. This case demonstrates some of the obstetric complications associated with congenital genital tract anomalies. PMID- 2562609 TI - Combined intrauterine and tubal ectopic pregnancy. PMID- 2562610 TI - Pulsatile ovarian fossa ectopic and the 'morning after' pill. PMID- 2562611 TI - The surgical treatment of tubal polyps. AB - Three of 5 women who had resection of intramural tubal polyps as part of their infertility treatment conceived normal pregnancies within 12 months. Linear salpingotomy for the resection of polyps may have a place in the surgical management of infertility. PMID- 2562612 TI - Generalized peritonitis due to spontaneous rupture of pyometra. AB - Spontaneous rupture of pyometra is a rare cause of generalized peritonitis. Only a few cases have been recorded in the English medical literature, most of which were associated with gynaecological malignancy. We have recently treated a patient with generalized peritonitis due to ruptured pyometra, in which there was no evidence of malignancy or other cervical disease. From comparison with the other published case reports, several features deserve emphasis. PMID- 2562613 TI - Treatment of ectopic pregnancy with intraamniotic methotrexate--a case report. PMID- 2562614 TI - An unusual malignancy arising in a benign cystic teratoma of the ovary. PMID- 2562616 TI - From Charcot to crystals: three vignettes. PMID- 2562615 TI - Is elective cerclage justified in the management of triplet and quadruplet pregnancy? AB - The main complication in multiple gestation with more than two fetuses is prematurity, with its concomitant increase in perinatal mortality and morbidity. Clearcut indications as to management of these pregnancies are lacking, and the efficiency of elective cerclage is controversial. Twelve triplets and three quadruplets out of 27 multiple pregnancies were electively sutured and compared to 10 triplets and two quadruplets without this procedure. Otherwise, both groups were managed uniformly regarding bed-rest, beta-mimetic drugs and dexamethasone for the enhancement of fetal lung maturity. Mean duration of pregnancy in patients with cerclage was 35 weeks, significantly longer than those who did not undergo this procedure (30.7 weeks) (p < 0.01). Furthermore, in the former group, the mean neonatal weight was significantly higher (p < 0.01), mean Apgar scores were better, significantly lower rates of respiratory distress syndrome occurred (p < 0.05), perinatal mortality rate was significantly reduced (p < 0.01) and the mean hospitalization period was shorter (p < 0.025). It seems that elective cervical suture is a definite contribution to the successful management of multiple pregnancies with more than two fetuses. PMID- 2562617 TI - Liver transplantation in Europe and changing patterns of liver care. PMID- 2562618 TI - Why me? The role of genes in human disease. PMID- 2562619 TI - Trends in medical litigation. PMID- 2562620 TI - Cancer chemotherapy 1990. PMID- 2562621 TI - RNA editing: a novel mechanism for the regulation of dietary cholesterol absorption. PMID- 2562622 TI - Mesmerism in the London medical circle. PMID- 2562623 TI - Caring for the elderly: yesterday, today, and tomorrow. PMID- 2562624 TI - Genes, bacteria and the key to arthritis. PMID- 2562625 TI - Modern anaesthesia--challenge and change. PMID- 2562626 TI - The annual oration. A backward look to the future. PMID- 2562627 TI - [Kidney transplantation: the current situation in Portugal, 1989]. AB - The modern treatment of renal failure has been one of the most exciting developments of modern medicine. Within this field the portuguese doctors had a performance that compares with the average developed countries. The number of patients that need haemodialysis and renal transplantation is still increasing. These treatments are very expensive. As the patients would die if not treated, this whole issue is a political concern and a public health problem. We give the demographic outline of this CRF patients group. Dialysis and transplantation should be in balance, with a particular stress on transplantation because it is much less expensive. Shortness of organ supply has been like elsewhere the most important limiting factor of renal transplantation. Some public controversy concerning brain death, permission, and commercial profit from organ donation, has been sometimes deleterious. We review the results of renal transplantation. In 1987 and 1988, 92% of transplanted kidneys had still function after one year. We conclude that our programs in Portugal can expand, on the condition that people keep their acceptance and give their support to transplanting teams. PMID- 2562628 TI - [Bone marrow transplantation: current perspectives]. AB - Bone marrow transplantation was introduced over thirty years ago in the treatment of several immuno-hematological conditions. During the last decade, there has been a substantial increase in the number of transplanted patients. However, results over the last five years have not significantly improved. Clinical indication conditioning regimens and related complications are reviewed and new developments pointed out. PMID- 2562629 TI - [State of the art in pediatric cardiology]. PMID- 2562630 TI - [State of the art in cardiology: realities and perspectives of ischemic cardiopathy in the 90's]. PMID- 2562631 TI - [Relationship between basic sciences and medicine: institutional, professional, and educational implications]. PMID- 2562632 TI - [State of the art in cardiothoracic surgery: evolution of thoracic surgery]. PMID- 2562633 TI - [State of the art in immuno-allergology: histamine-releasing factors (HRF)]. AB - The present concepts on histamina releasing factors are discussed, their cellular origins, their effect on the target cells and its importance in asthma immunopathology. Personal results on the production of HRF by alveolar macrophages stimulated by allergen and its action on non atopic basophils are presented. Alveolar macrophages from atopic asthmatics release HRF in presence of allergen. Macrophage from non atopic do not. HRF has been tested in a Human Basophil Degranulation Test (HBDT) preparation, a significant degranulation induced by HRF from atopic patient being only observed when atopic basofils are used in HBDT. These results confirm that allergen could induce HRF production from alveolar macrophages and that this HRF could degranulate basophils in atopic and induce mediators release. PMID- 2562634 TI - [State of the art in plastic and reconstructive surgery: a growing art]. AB - On studying the state of the art in Plastic and Reconstructive Surgery there are two different aspects that have achieved very different levels of advance and progress: the organisation and establishment of this speciality, and scientific and technical progress. In an analysis of the former. The author outlines the principal problems facing Plastic Surgery in Portugal, the present shortcomings and the almost complete lack of coherent planning, and the prevailing rules for the training of new specialists. A study of the second aspect reveals manifest progress and innovation in many areas of this speciality. The Author describes in greater detail, controlled tissue growth, innovations with flaps, principally facial-cutaneous and inverted flow, advances in the remodelling of the face where there are many examples of implantation of material, modelling osteotomy and surgical interventions in the field of muscular dynamics, and lipo-aspiration which has extended horizons in both Plastic and Reconstructive Surgery. PMID- 2562635 TI - [State of the art in otology]. PMID- 2562636 TI - [State of the art in rheumatology: various historical notes]. PMID- 2562637 TI - [Impact of information technology on the classic environment: short- and long term perspectives in Portugal and the European arena]. PMID- 2562638 TI - Clinical applications of SPECT with special reference to oncology. PMID- 2562639 TI - Peer review: another delicate balance. PMID- 2562640 TI - Efficacy and toxicity elicited by recombinant interferons alpha and gamma when administered in combination to tumor-bearing mice. AB - Administration of rHuIFN-alpha A/D and rMuIFN-gamma as single agents to tumor bearing mice resulted in a dose-related antitumor effect in each of the six models studied. When the IFNs were given in combination, the effects varied between the tumor systems. No increase in efficacy was seen in mice bearing B16 F10 melanoma or M5076 reticulum cell sarcoma while additive antitumor activity was shown in the KA31 fibrosarcoma and P388 leukemia systems. Mice inoculated with L1210 lymphoma or colon 38 carcinoma, however, revealed enhanced efficacy which was greater than additive. The data also reveal that combination of IFNs alpha and gamma administered to normal and tumor-bearing mice resulted in toxicity which was not predicted by the appropriate doses of the single agents. These studies suggest that combination of IFNs alpha and gamma may provide greater therapeutic utility than the single agents and underscore the need for additional, carefully designed preclinical and clinical efforts. PMID- 2562641 TI - Effects of purified recombinant human interleukin-3 on the growth of human hematopoietic progenitor cells in "long-term" bone marrow culture. AB - Recombinant human interleukin-3 (rhuIL-3) was assessed for its effects on the growth of normal human hematopoietic bone marrow nucleated cells, and on granulocyte-macrophage (CFU-GM) and erythroid (BFU-E) progenitor cells in a liquid culture system which allows for the prolonged growth of these cells in vitro. RhuIL-3, at concentrations of 100 and 500 units/mL, significantly enhanced the numbers of nucleated cells, as well as the numbers of supernatant and adherent CFU-GM and BFU-E growing in tissue culture flasks or dishes over a period of 4 to 6 weeks. The results demonstrated the rhuIL-3 has a stimulating effect on the growth of human marrow cells in prolonged culture. This information is consistent with the effects of rhuIL-3 in short-term marrow colony assays in vitro and with the in vivo actions of recombinant murine IL-3 in mice, and may be of relevance to clinical trials that will be assessing the hematopoietic effects of rhuIL-3 in humans. PMID- 2562642 TI - Interleukin-4 (B-cell stimulatory factor-1) augments the in vivo generation of cytotoxic cells in immunosuppressed animals. AB - Thymus-derived lymphocytes (T cells) are thought to play an important role in the recognition and destruction of neoplastic cells in the host. This principle has provided a foundation for the establishment of therapy with T-cell-stimulating lymphokines, notably interleukin-2, as an approach to the eradication of certain malignancies. Another lymphokine, B-cell-stimulatory factor-1 (BSF-1), also known as IL-4, has also been shown to be capable of inducing T-cell proliferation and cytolytic activity in vitro. We demonstrate herein that in immunosuppressed mice, in vivo IL-4 administration enhances the ability of treated animals to generate cytotoxic T lymphocytes directed against an allogeneic tumor challenge. Moreover, IL-4 is approximately 25 times more effective, on a weight basis, than is IL-2 in augmenting cytotoxic T-lymphocyte activity. This difference in efficiency between the two lymphokines may be partly due to the in vivo half-life. We have found that IL-4 has a serum half-life of 19 +/- 2 min following intravenous administration, in contrast to the half-life of IL-2, which has been reported to be 3.7 min +/- 0.8. These results are not only of interest for our basic understanding of the physiological role of IL-4 but may have immediate importance in clinical settings where lymphokine therapy is contemplated. PMID- 2562643 TI - Influence of purified recombinant human macrophage colony stimulating factor in mice. AB - The endotoxin-resistant strain of mouse, C3H/Hej, was assessed for hematological responsiveness to multiple injections of high dosages of purified recombinant human macrophage colony stimulating factor (rhu-M-CSF). Mice were administered the rhu M-CSF i.p. at dosages of 40 micrograms per injection, 2 or 3 times per day for 4 days. This resulted in significant increases in circulating leukocytes compared to control mice given sterile pyrogen-free saline. Assessment of the marrow and spleen of these mice on the 5th day noted a significant reduction in the numbers of marrow hematopoietic progenitor cells, with no change in their cycling rates. In contrast, splenic granulocyte-macrophage and erythroid progenitor cell numbers were markedly increased and the cycling rates of these progenitors plus those of multipotential progenitors were significantly enhanced. Marrow and splenic early myeloid cells (blasts, promyelocytes, and myelocytes) and macrophages were increased, while marrow and splenic PMN were decreased. The results suggest that multiple injections of high dosages of rhu-M-CSF to previously untreated mice for a short period of time has a modest enhancing effect on blood leukocyte levels. This is associated with a shift of hematopoietic cell activity from the marrow to the spleen. PMID- 2562645 TI - Potential therapeutic value of muramyl peptides for modulating human immunologic responses. AB - The clinical potential and the limitations of synthetic muramyl peptides have been suggested through extensive work describing their immunomodulating properties and toxicology. The intent of this paper is to offer the clinician a summary of these studies and to introduce the reader to the biological effects associated with administration of muramyl peptides. PMID- 2562646 TI - Disturbing homeostasis: recent results of ongoing immunotherapy trials at the NCI. PMID- 2562644 TI - The diabetic rat as an impaired wound healing model: stimulatory effects of transforming growth factor-beta and basic fibroblast growth factor. AB - Two models of wound repair compared the effect of defined, recombinant growth factors on the rate of wound repair in both normal and streptozotocin-induced diabetic rats: subcutaneous implantation of polyvinyl alcohol sponges and incisional wounding. Transverse incisional wounds were made on the dorsal surface of rats and closed with steel sutures. Three days postwounding the rats received a single injection of either transforming growth factor-beta or vehicle alone directly into the wound site. Animals were sacrificed 7, 14, and 21 days postwounding, and fresh and formalin-fixed wound tensile strength were measured. Diabetic rats had expected defects in wound repair, including decreased granulation tissue and reduced amounts of collagen, protein, and DNA. Fresh tensile strength of the diabetic incisions was 53% of normal on Day 7 (p < or = .01) and 29% of normal on Day 21. Fixed tensile strength was 41% of normal on Day 7 (p < or = .01) and fell to 78% of normal by Day 21 (p < or = .01), suggesting that collagen concentrations of diabetic wounds increased towards normal but did not undergo maturation. TGF beta produced a moderate increase in tensile strength of fresh and fixed wounds of diabetic rats, but not to the levels of wounds in untreated normal rats. Sponges fill with granulation tissue, their reproducible rate of organization being measured by histological and biochemical methods. A single injection into sponges 3 days postimplantation of basic fibroblast growth factor, transforming growth factor-beta, or vehicle only, was evaluated at 7 and 9 days postimplantation. In the sponge model, bFGF and TGF beta were each able to induce significant increases in the accumulation of granulation tissue in both diabetic and normal rats. TGF beta increased the collagen content of sponges by 136% in sponges from diabetic animals (p < or = .001), thereby raising the collagen content to that of normal control wounds, while stimulating a 49% (p < or = .02) increase in sponges from normal animals on Day 9. By contrast, the response to bFGF was predominantly an increase in the protein and DNA content of the sponges. These results emphasize the differential effects of the two cytokines in accelerating healing under conditions of defective wound repair. PMID- 2562647 TI - Active specific immunotherapy of melanoma: clinical results and correlates. PMID- 2562648 TI - Prospects for gene therapy. PMID- 2562649 TI - Present status and future of monoclonal antibodies. PMID- 2562650 TI - Absorption of human growth hormone from the rat lung. AB - Recombinant methionyl human growth hormone (hGH) was administered intratracheally to adult rats, and serum concentrations of immunoreactive hGH were measured for up to 24 h. The mean absolute bioavailability was approximately 36% after 18h and was similar for doses of 0.75, 1.5, and 3 mg/kg. Peak serum hGH concentrations occurred at approximately 6 h after dosing. Tritiated hGH (3H-hGH) was used to follow the clearance of hormone from the lungs. Disappearance was linear with time and by 24 h approximately 70% of the radioactivity was gone from the lungs (elimination half-life = approximately 10.5h). Monomeric and aggregated hGH appeared to account for the majority of the residual 30% of radioactivity. Immunohistochemical localization of hGH in the alveoli suggested that the hormone was concentrated in a thin layer at the air-epithelial boundary. Pulmonary macrophages, which also stained for hGH, probably degrade hGH and thus account for some loss of material in the lungs. These studies suggest that the lung may be an alternative route for systemic delivery of recombinant proteins which are currently delivered by injection. PMID- 2562651 TI - A phase I/II study of recombinant tumor necrosis factor and recombinant interferon gamma in patients with AIDS-related complex. AB - A clinical trial was conducted to determine the tolerance and toxicity of recombinant tumor necrosis factor (rTNF) and recombinant interferon gamma (rIFN gamma) when administered concurrently by continuous intravenous infusion to 11 patients with the AIDS-related complex (ARC). In addition, HIV culture, p24 antigen levels, and CD4 positive lymphocytes were monitored to obtain preliminary evidence of antiviral and immunologic effects. Two 5-day treatment cycles were separated by a 9-day washout period. Two patients were entered at each dosage level and each patient received the two 5-day treatment cycles at two sequential dose levels ranging from 1 to 25 micrograms/m2. Two patients did not complete their second treatment cycle--one due to the development of a rash, the second due to central venous catheter discomfort. The occurrence of phlebitis with peripheral vein administration of these agents necessitated administration via central venous catheter. With the exception of a single patient who developed severe headache at the 25 micrograms/m2 dose, severe clinical toxicities were not observed. Fever, chills, headache, and myalgias were the most significant clinical toxicities observed and all were dose dependent. The percentage fall in total granulocytes was dose dependent and ranged from 17% at the 1 microgram/mm2 dose to 48% at both the 15 and 25 micrograms/mm2 dose levels. The mean nadir granulocyte count was 1694/mm3. No significant renal or hepatic toxicity was observed. Of 22 treatment cycles the CD4 cell number was increased in 11, unchanged in 7, and decreased in 4. The mean CD4 cell number did not change significantly (176 +/- 143/mm3 pretherapy versus 279 +/- 305/mm3 posttherapy).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562652 TI - Epidermal growth factor: physiological roles and therapeutic applications. PMID- 2562653 TI - Tissue-type plasminogen activator: mutants, variants, and adjunctive treatment. PMID- 2562654 TI - Human granulocyte-macrophage colony-stimulating factor. PMID- 2562655 TI - Antimicrobial activity of synthetic bactenecin. AB - Bactenecin is an antimicrobial peptide isolated from bovine neutrophils. Bactenecin was synthesized by solid-phase peptide synthesis and renatured to a fully disulfide bonded form. The peptide inhibits the growth of Escherichia coli and Staphylococcus aureus at the same concentration reported for the peptide purified from bovine neutrophils. Bactenecin inhibits the growth of other medically important bacteria and yeast, and it kills the fungus Trichophyton rubrum. Acetylation and amidation of the amino- and carboxy-termini of bactenecin do not change its potency, while replacement of its two cysteine residues with serine decreases the potency. PMID- 2562656 TI - Direct effects of IL-4 on the in vitro differentiation and proliferation of hematopoietic progenitor cells. AB - The purpose of this study was to analyze the effects of recombinant human interleukin 4 (IL-4) on the differentiation and proliferation in vitro of human granulocyte/macrophage (GM) and erythroid progenitors. IL-4 was added to either fetal bovine serum (FBS)-supplemented or to FBS-deprived cultures of unfractionated human marrow cells or marrow cells depleted of adherent and/or T cells. Paradoxical effects similar to those reported in the murine system were detected in these experiments. In FBS-supplemented cultures, IL-4, which had no effect on the growth or erythroid bursts (from burst-forming cells; BFU-E) detected in the presence of Epo alone, decreased by 46% the number of erythroid bursts detected in the presence of Epo and phytohemagglutinin-stimulated leukocyte-conditioned medium (PHA-LCM). In contrast, in FBS-deprived cultures, IL 4 increased by 30-700% the number of erythroid bursts in cultures containing Epo alone or containing Epo, IL-3, and GM-CSF. The stimulatory effect of IL-4 on erythroid burst growth under FBS-deprived conditions was particularly evident when adherent cells were removed. Under the conditions investigated, IL-4 had little effect on the growth of GM colonies. In FBS-deprived suspension cultures of nonadherent, T-cell-depleted marrow cells, IL-4 maintained both the number of BFU-E and CFU-GM for at least 8 days. In these cultures, IL-4 antagonized the capacity of IL-3 to increase the number of BFU-E but IL-4 and IL-3 acted together to maintain the number of CFU-GM. To determine if IL-4 acted directly or indirectly, its effects on the growth of factor-dependent subclones of the murine progenitor cell line 32D were analyzed. Three subclones were studied: the original IL-3-dependent clone 32D cl.3, the Epo-dependent erythroid clone 32D Epo 1, and the G-CSF-dependent myeloid clone 32D G-1. IL-4 alone failed to induce colony growth from these cell lines. However, IL-4 inhibited by 25% the number of colonies formed by 32D cl.3 in the presence of IL-3 while increasing by 25% and 25-50% the number of colonies formed by 32D Epo-1 and 32D G-1 in the presence of Epo or G-CSF, respectively. These results indicate that human IL-4, as its murine counterpart, is a multilineage growth factor with paradoxical effects which are mediated by the direct action of IL-4 on progenitor cells. PMID- 2562657 TI - The role of cytokines in autoimmunity. PMID- 2562658 TI - Stem cells: the generation and maintenance of cellular diversity. PMID- 2562659 TI - The oral-aboral axis of a sea urchin embryo is specified by first cleavage. AB - Several lines of evidence suggest that the oral-aboral axis in Strongylocentrotus purpuratus embryos is specified at or before the 8-cell stage. Were the oral aboral axis specified independently of the first cleavage plane, then a random association of this plane with the blastomeres of the four embryo quadrants in the oral-aboral plane (viz. oral, aboral, right and left) would be expected. Lineage tracer dye injection into one blastomere at the 2-cell stage and observation of the resultant labeling patterns demonstrates instead a strongly nonrandom association. In at least ninety percent of cases, the progeny of the aboral blastomeres are associated with those of the left lateral blastomeres and the progeny of the oral blastomeres with the right lateral ones, respectively. Thus, ninety percent of the time the oral pole of the future oral-aboral axis lies 45 degrees clockwise from the first cleavage plane as viewed from the animal pole. The nonrandom association of blastomeres after labeling of the 2-cell stage implies that there is a mechanistic relation between axis specification and the positioning of the first cleavage plane. PMID- 2562661 TI - Detection of age-related changes in the distributions of keratan sulfates and chondroitin sulfates in developing chick limbs: an immunocytochemical study. AB - A panel of four separate monoclonal antibodies, all known to specifically recognize epitopes on keratan sulfate glycosaminoglycans, were employed in an immunocytochemical study of developing chick hind limbs. In addition, two monoclonal antibodies specific for epitopes on chondroitin/dermatan sulfate glycosaminoglycans were employed on equivalent sections to determine the degree of colocalization of keratan and chondroitin/dermatan sulfates. The spatial distributions of keratan sulfate and chondroitin/dermatan sulfate differed to some extent. In younger embryos, high extracellular concentrations of keratan sulfate occurred in joints and articular cartilages, with diminishing amounts being present in epiphyseal and diaphyseal regions. The high concentration of keratan sulfate in joints and articular cartilage corresponded to equally high concentration of chondroitin-6 sulfate. With advancing age, the above mentioned distribution was modified, most notably by increased amounts of keratan sulfate within diaphyseal regions. Finally, the use of four different anti-keratan sulfate monoclonal antibodies made it possible to compare keratan sulfate epitope expression. Differences in keratan sulfate epitopes were noted in some regions of bones, mostly in diaphyseal regions of younger bones and epiphyseal regions of older bones. This pattern of keratan sulfate expression suggests that different types of keratan sulfate may be present and their expression may be developmentally regulated. PMID- 2562660 TI - Prepupal differentiation in Drosophila: distinct cell types elaborate a shared structure, the pupal cuticle, but accumulate transcripts in unique patterns. AB - The components of the pupal cuticle are the main differentiation products synthesized by both the larval and adult epidermis during the prepupal period of Drosophila development. The pupal cuticle is formed in vitro by imaginal discs in response to a 6 h pulse of 20-hydroxyecdysone (20-HE). We previously described the isolation and initial characterization of four ecdysone-dependent genes (EDGs) whose expression in imaginal discs occurs only in response to a pulse of 20-HE. In this report, we demonstrate that the pattern of temporal and tissue specific expression of these EDGs in vivo is like that expected for genes that encode pupal cuticle proteins. Transcripts of these genes are detected in prepupae only in the epidermis and only when cuticle components are synthesized and secreted. Nonetheless, their temporal and spatial patterns of accumulation differ. EDG-84A-1 transcripts accumulate only in prepupae and only in imaginal cells. EDG-78E and EDG-64CD transcripts accumulate at the same time in both larval and imaginal cells. EDG42-A transcripts appear first in prepupae in imaginal cells and then, after a 2-4 h lag, in larval cells. It is evident that some genes are not restricted in their expression to only larval or imaginal epidermis. PMID- 2562662 TI - Expression of N-CAM precedes neural induction in Pleurodeles waltl (urodele, amphibian). AB - The appearance and localization of N-CAM during neural induction were studied in Pleurodeles waltl embryos and compared with recent contradictory results reported in Xenopus laevis. A monoclonal antibody raised against mouse N-CAM was used. In the nervous system of Pleurodeles, it recognized two glycoproteins of 180 and 140x10(3) M(r) which are the Pleurodeles equivalent of N-CAM-180 and -140. Using this probe for immunohistochemistry and immunocytochemistry, we showed that N-CAM was already expressed in presumptive ectoderm at the early gastrula stage. In late gastrula embryos, a slight increase in staining was observed in the neurectoderm, whereas the labelling persisted in the noninduced ectoderm. When induced ectodermal cells were isolated at the late gastrula stage and cultured in vitro up to 14 days, a faint polarized labelling of cells was observed initially. During differentiation, the staining increased and became progressively restricted to differentiating neurons. PMID- 2562663 TI - Mouse neural crest cells secrete both urokinase-type and tissue-type plasminogen activators in vitro. AB - Neural tubes of E8-5 day mouse embryos were dissected and cultured in serum substitute-supplemented medium to allow the emigration of neural crest cells. After 48 h of culture the neural tubes were removed. The neural crest cells were then cultured for 12 h in serum-free medium, and their culture supernatant was studied by electrophoresis and zymography. The cultured cells were shown to secrete both urokinase-type and tissue-type plasminogen activators. When the truncal neural tube was divided in four equal segments, the secretion pattern of the two types of plasminogen activators was similar for the cells from the three most anterior segments; cells having migrated from the most caudal one, i.e. consisting of the neural plate, secreted a higher level of urokinase-type plasminogen activator. The secretion in vitro of plasminogen activators by neural crest cells is in accord with the postulated importance of these proteases in cellular migration. PMID- 2562664 TI - Retinoic acid application to chick wing buds leads to a dose-dependent reorganization of the apical ectodermal ridge that is mediated by the mesenchyme. AB - Local application of retinoic acid to wing buds of chick embryos leads to dose- and position-dependent changes in the pattern of cellular differentiation. Early effects of retinoid treatment on the apical ectodermal ridge coordinate pattern changes and morphogenesis. The length of the apical ridge increases when additional digits will form but decreases when digits are lost. These changes in length can be understood in terms of a threshold response to the local retinoid concentration that results in either disappearance or maintenance of the ridge (Lee & Tickle, J. Embryol. exp. Morph. 90, 139-169 (1985)). Here, we have analysed the mechanisms involved in ridge disappearance by locally applying retinoic acid to the apex of stage 20 chick wing buds. With this treatment regime, low doses give duplicated digit patterns and higher doses truncations. The height of the apical ridge is progressively reduced with increasing doses of retinoid and the time course of ridge flattening indicates that the height of the ridge is correlated with bud outgrowth. With high doses of retinoic acid, the typical ridge, a pseudostratified epithelium in which the columnar cells are tightly packed, disappears and the epithelium at the tip of the bud consists of loosely packed cuboidal cells. Shortly after treatment, there is a decrease in the number of gap junctions between ridge cells. This early change in cell contacts suggests that gap junctions may be involved in maintaining epithelial morphology. When treated epithelium is recombined with untreated mesenchyme, an apical ridge is reestablished and distal structures can be generated. In contrast, when treated mesenchyme is recombined with the epithelium from normal buds, only proximal structures are formed. Therefore, retinoids can lead to a reorganization of the apical ectodermal ridge which is mediated and maintained by the mesenchyme. PMID- 2562665 TI - Melanogenesis of avian neural crest cells in vitro is influenced by external cues in the periorbital mesenchyme. AB - Avian melanoblast differentiation was studied by explantation of the neural tube and periorbital mesenchyme. Outgrowths from the mesenchymal explants consisted of a mixed population of melanocytes, melanoblasts and fibroblasts, whilst typical neural crest populations migrated from the neural tube explants. Cells that differentiated within explants of mesenchyme, produced elongate black eumelanosomes of normal ultrastructure which were identical to those found in the ocular connective tissues. However, melanoblasts that differentiated within outgrowths of mesenchyme or neural tube produced round brown melanosomes of highly abnormal ultrastructure. Some of these melanosomes contained a few disorganised melanosomal filaments whilst others had granular melanin with complete absence of filaments. This abnormality of phenotype was invariant over a range of culture conditions that modified cell behaviour, the timing of differentiation and the abundance of the pigmented cells. These experiments suggest that local factors in the mesenchyme are essential for the induction of melanogenesis in the presumptive connective tissue melanocyte. PMID- 2562666 TI - Glial interactions with neurons during Drosophila embryogenesis. AB - A monoclonal antibody (Mab5B12) demonstrating specificity for glial cells within the central and peripheral nervous systems of Drosophila has been used in combination with neural-specific antibodies to study the early organization of the Drosophila embryo. The embryonic central nervous system of Drosophila contains cells within the ventral midline that are recognized by monoclonal antibody 5B12. These cells are not recognized by either a polyclonal antiserum to horse radish peroxidase, which recognizes several antigens on the surface of Drosophila neurons, or Mab22C10, which recognizes an antigen specific to the peripheral nervous system. Mab5B12-positive cells lie dorsal both to the developing anterior and posterior commissures in each thoracic and abdominal segment and to the supraoesophageal commissure. They ensheath these commissures in later stage embryos. Other Mab5B12-positive cells lie dorsolateral to the CNS and send processes laterally to the lateral sensilla during axonogenesis in the PNS. These cells surround the axons of the intersegmental and segmental nerves. Other cells that line the advancing ectoderm during dorsal closure and surround the anal pads also express the Mab5B12 antigen. Neuronal cell cultures derived from Drosophila gastrulae contain cells expressing the Mab5B12 antigen. These cells can be found separate or in close association with neuronal clusters and their axons. PMID- 2562667 TI - A premature acrosome reaction is programmed by mouse t haplotypes during sperm differentiation and could play a role in transmission ratio distortion. AB - Mouse t haplotypes are variant forms of chromosome 17 that can be transmitted at non-Mendelian ratios by heterozygous +/t males. The accumulated genetic data indicate that '+-sperm' and 't-sperm' are produced in equal numbers but that most '+-sperm' are rendered dysfunctional, so that 't-sperm' have a relative advantage at fertilization. To date, the basis for this t-induced sperm dysfunction has remained unknown. Here we demonstrate that a high proportion of sperm obtained from certain strains of +/t mice undergo a premature acrosome reaction under in vitro capacitation conditions. The simplest interpretation of these data, in conjunction with previous results, is that developing '+-spermatids' are preprogrammed by 't-spermatids' to undergo this premature reaction. Since acrosome-reacted sperm are unable to participate in the process of fertilization, this defect could account for the extreme distortion of transmission ratio observed from mice heterozygous for a class of complete t haplotypes. PMID- 2562668 TI - Localized synthesis of the Vg1 protein during early Xenopus development. AB - The Xenopus Vg1 gene encodes a maternal mRNA that is localized to the vegetal hemisphere of both oocytes and embryos and encodes a protein related to the TGF beta family of small secreted growth factors. We have raised antibodies to recombinant Vg1 protein and used them to show that Vg1 protein is first detected in stage IV oocytes and reaches maximal levels in stage VI oocytes and eggs. During embryogenesis, Vg1 protein is synthesized until the gastrula stage. The embryonically synthesized Vg1 protein is present only in vegetal cells of an early blastula. We find that Vg1 protein is glycosylated and associated with membranes in the early embryo. Our results also suggest that a small proportion of the full-length Vg1 protein is cleaved to give a small peptide of M(r) = approximately 17 x 10(3). These results support the proposal that the Vg1 protein is an endogenous growth-factor-like molecule involved in mesoderm induction within the amphibian embryo. PMID- 2562669 TI - Immunohistochemical localisation of chondroitin sulphate proteoglycans and the effects of chondroitinase ABC in 9- to 11-day rat embryos. AB - Studies on cell behaviour in vitro have indicated that the chondroitin sulphate proteoglycan (CSPG) family of molecules can participate in the control of cell proliferation, differentiation and adhesion, but its morphogenetic functions had not been investigated in intact embryos. Chondroitin/chondroitin sulphates have been identified in rat embryos at low levels at the start of neurulation (day 9) and at much higher levels on day 10. In this study we have sought evidence for the morphogenetic functions of CSPGs in rat embryos during the period of neurulation and neural crest cell migration by a combination of two approaches: immunocytochemical localization of CSPG by means of an antibody, CS-56, to the chondroitin sulphate component of CSPG, and exposure of embryos to the enzyme chondroitinase ABC. Staining of the CS-56 epitope was poor at the beginning of cranial neurulation; bright staining was at first confined to the primary mesenchyme under the convex neural folds late on day 9. In day 10 embryos, all mesenchyme cells were stained, but at different levels of intensity, so that primary mesenchyme, neural crest and sclerotomal cells could be distinguished from each other. Basement membranes were also stained, particularly bright staining being present where two epithelial were basally apposed, e.g., neural/surface ectoderms, dorsal aorta/neural tube, prior to migration of a population of cells between them. Staining within the neural epithelium was first confined to the dorsolateral edge region, and associated with the onset of neural crest cell emigration; after neural tube closure, neuroepithelial staining was more general. Neural crest cells were stained during migration, but the reaction was absent in areas associated with migration end-points (trigeminal ganglion anlagen, frontonasal mesenchyme). Embryos exposed to chondroitinase ABC in culture showed no abnormalities until early day 10, when cranial neural crest cell emigration from the neural epithelium was inhibited and neural tube closure was retarded. Sclerotomal cells failed to take their normal pathway between the dorsal aorta and neural tube. Correlation of the results of these two methods suggests: (1) that by decreasing adhesiveness within the neural epithelium at specific stages, CSPG facilitates the emigration of neural crest cells and the migratory movement of neuroblasts, and may also provide increased flexibility during the generation of epithelial curvatures; (2) that by decreasing the adhesiveness of fibronectin-containing extracellular matrices, CSPG facilitates the migration of neural crest and sclerotomal cells. This second function is particularly important when migrating cells take pathways between previously apposed tissues. PMID- 2562670 TI - Molecular characteristics of cytostatic factors in amphibian egg cytosols. AB - In amphibians, zygotes microinjected with cytosol of unactivated eggs are arrested at metaphase of mitosis. The factor responsible for this effect has been designated 'cytostatic factor, (CSF)'. CSF is inactivated by Ca2+ addition to cytosols. During storage of the Ca(2+)-containing cytosols, a stable CSF activity develops. Therefore, the first Ca(2+)-sensitive CSF and the second Ca(2+) insensitive CSF have been referred to as primary CSF (CSF-1) and secondary CSF (CSF-2), respectively. We have partially purified CSF-1, which had been stabilized with NaF and ATP, and CSF-2 from cytosols of Rana pipiens eggs by ammonium sulphate (AmS) precipitation and sucrose density gradient centrifugation or gel filtration, and investigated their molecular characteristics. CSF-1 was sensitive to protease, but resistant to RNAse, and inactivated within 2 h at 25 degrees C. CSF-1 could be sedimented in a sucrose density gradient from a fresh cytosol or its crude fraction precipitated at 20-30% saturation of AmS, showing the sedimentation coefficient 3S. When analyzed by SDS-polyacrylamide gel electrophoresis (PAGE), all the proteins in partially purified CSF-1 samples entered the gel and were separated into numerous peptide bands. In contrast, CSF 2 was an extremely large molecule, being eluted from Sepharose columns as molecules larger than 2 x 10(6), and failed to enter the gel when analyzed by SDS PAGE. It could be purified 40 times from cytosols. CSF-2 was a highly stable molecule, being neither inactivated nor dissociated at pH 11.5 or by 4M-NaCl and LiCl and 8 M-urea. It was also resistant to RNAse treatment. However, CSF-2 could be broken down into small peptides of variable sizes by trypsin, alpha chymotrypsin, and papain, but not by S. aureus V8 protease, although it was less sensitive to proteases than CSF-1. The dose-dependency test showed that the activity of CSF-2 is independent of its concentration and that an amount of CSF-2 could cause cleavage arrest earlier when injected into a blastomere in a larger volume. PMID- 2562671 TI - A vital dye analysis of the timing and pathways of avian trunk neural crest cell migration. AB - To permit a more detailed analysis of neural crest cell migratory pathways in the chick embryo, neural crest cells were labelled with a nondeleterious membrane intercalating vital dye, DiI. All neural tube cells with endfeet in contact with the lumen, including premigratory neural crest cells, were labelled by pressure injecting a solution of DiI into the lumen of the neural tube. When assayed one to three days later, migrating neural crest cells, motor axons, and ventral root cells were the only cells types external to the neural tube labelled with DiI. During the neural crest cell migratory phase, distinctly labelled cells were found along: (1) a dorsolateral pathway, under the epidermis, as well adjacent to and intercalating through the dermamyotome; and (2) a ventral pathway, through the rostral portion of each sclerotome and around the dorsal aorta as described previously. In contrast to those cells migrating through the sclerotome, labelled cells on the dorsolateral pathway were not segmentally arranged along the rostrocaudal axis. DiI-labelled cells were observed in all truncal neural crest derivatives, including subepidermal presumptive pigment cells, dorsal root ganglia, and sympathetic ganglia. By varying the stage at which the injection was performed, neural crest cell emigration at the level of the wing bud was shown to occur from stage 13 through stage 22. In addition, neural crest cells were found to populate their derivatives in a ventral-to-dorsal order, with the latest emigrating cells migrating exclusively along the dorsolateral pathway. PMID- 2562672 TI - Development and phenotypic variability of genetically identical half mouse embryos. AB - The growth and development of three groups of genetically identical F1 C57BL/6J female x SJL/J male mice were compared to examine whether embryo manipulation affects subsequent postnatal growth and development of mammalian embryos: (1) controls--the natural offspring of timed matings, (2) transferred controls- offspring from 2-cell embryos transferred to recipients 1 day asynchronous, and (3) transferred half embryos--offspring developing from one blastomere from the 2 cell stage transferred to recipients 1 day asynchronous. The recipients were C57BL/6J females. No differences were found in the age at eye opening and vaginal opening. At 5 days after birth the median body weights of the controls were lower than the weights of the transferred groups. This result could be explained by the larger litter size in the control group. The overall variances of the body weights did not differ between the groups. By the second week after birth a marked increase in overall variances of body weights of the transferred groups, compared with the control group, was observed. At 5 days after birth, the median tail lengths did not differ between groups, and overall variances were the same. By the second week, the overall variances of the tail lengths of the transferred groups were significantly greater than that of the control group. Possibly the increased overall variances of the body weight and the tail length of the transferred groups are related to the smaller litter size in these groups which affects competition for food and the ambient temperature in the nest. The overall results suggest newborn mice that have developed from half embryos have compensated for their initial deficiency. The intraclass correlation coefficients for body weight and tail length are approximately the same in all groups. Thus, producing artificial identical twins by embryo bisection may not affect their potential usefulness in the design of experiments. PMID- 2562673 TI - Trends and current status in childhood mortality. PMID- 2562674 TI - [Science and social activities--passions of her life]. PMID- 2562675 TI - [Consideration of nuclear magnetic resonance imaging (NMR) in neuroradiology]. AB - The purpose of the study was to present, on the basis of literature data, the possibility of examination of the central nervous system by means of magnetic nuclear resonance. Advantages and limitations of the method are described in cases of brain tumours, white matter diseases, parasitic diseases and vascular changes, inflammatory processes as well as dys-histogenesis. The main advantage of the method which will make its future use particularly wide-spread is the fact that it eliminates exposure to ionizing radiation, makes possible determination of the character of the tissue based on the content of hydrogen nucleus in a given area, and enables differentiation to be made of the tissues with different interatomic bonds, that is those which contain different chemical compounds. Diagnostic potential of computed tomography, ultrasonography, magnetic nuclear resonance, emission tomography and subtraction angiography have been compared. The hazards associated with clinical application of NMR technique are also discussed. PMID- 2562676 TI - [The basilar artery of humans]. AB - The basilar artery in 44.4% of the cases came into being by joint of two vertebral arteries on the height of the inferior margin of the pont. However, in 40.4%--below, and in 15.1%--above of this margin. In 78.7% of the human the course of basilar artery was rectilinear. In 20.5% of the cases the bulge of this artery was observed on the right or left side, whereas in 0.8%--the S-shaped forms. In 56% of the persons the segmentation of the basilar artery on the posterior cerebral arteries takes place on the height of the superior margin of the pont, in 32.9%--above, and in 11.1%--below this one. The results of the investigations concerning the place of genesis, the height of segmentation and the direction of course of the particular artery, were permitted on separation of nine types of these blood vessels. The diameter of the initial and terminal section of the basilar artery at the new-born children was--respectively--1.9 and 1.8 mm, whereas at the adult persons it exceeded 4 mm. The length of this vessel ranged from 17.2 mm in the above-mentioned children to approximately 33 mm in persons over 20 years old. In all the examined cases the basilar artery was passing the arteries of the pont: medial and lateral and almost always the superior and the inferior anterior cerebellar arteries. In 12.4% of the persons the inferior posterior cerebellar arteries and, moreover, in 22.2%--the labyrinth arteries were beginning from this blood vessel. PMID- 2562677 TI - The influence of bromocriptine on sexual activity in ethanol-exposed male rats. AB - Long-lasting intake of ethanol and its influence on certain changes in the activity of male sexual hormones is the subject of numerous experiments. It is suggested that the decreased libido, hypotrophy of testicles and impotence in individuals subjected to ethanol, are connected with the lowered level of testosterone. Various mechanisms may be responsible for such an activity of ethanol. There has been pointed out that long-lasting intake of ethanol quickens the metabolism of testosterone in the liver. The results of other experiments indicate that ethanol and/or its metabolite, acetaldehyde decrease the biosynthesis of testosterone in the Leydig cells. It has been proved, too, that ethanol reduces the number of testicular gonadotropin receptors what, additionally, decreases the synthesis of testosterone. Another reason for the disordered function of gonads in alcoholics may be the influence of ethanol on the central nervous system structures. Particularly, it refers to the neurones of hypothalamus because they can regulate excretion of the hormones of anterior pituitary gland. In this way, hypothalamus may affect not only the level of the hormones produced by gonades but also to release pituitary prolactin. The present state of research indicates that ethanol results in clear changes in the function of the dopaminergic system of the brain. The influence of ethanol upon the neurones of this system depends on its dose and the period of activity. Most researches point out that prolonged administration of ethanol leads to hypofunction of dopaminergic neurones of the brain. PMID- 2562678 TI - Retroperitoneal tumors. AB - Retroperitoneal tumors (RPTs) are most commonly found in anatomically confined spaces. Primary RPTs are to be distinguished from those arising from retroperitoneal organs or from the metastatic masses. They may originate either from anyone of the three embryonal leaves or may take on more complex composition, consequently accounting for their varied histological appearance (1, 10). Badowski et al. (1) gave the following occurrence ratio: mesenchymal tumors account for 61.5%, mixed tumors for 23% and nervous tissue tumors--15.5%. They possess either benign or malignant potentials, with invasion of the neighbouring tissues and organs by the growth. The mode of disease at early stage is rarely conspicuous. Patients usually present with vague abdominal pains and palpable resistance. The techniques that are employed consist in the CT, USG and the selective angiography accompanied by passage films and the urography. PMID- 2562679 TI - [Statistical analysis of body and lung mass of animals subjected to a single experimental insufflation of soil dust and electro-energetic ashes]. AB - The studies were performed on 44 white female Wistar rats which were intratracheally administered the suspension of the soil dust and the electro energetic ashes. The electro-energetic ashes were collected from 6 different local heat and power generating plants while the soil dust from several random places of our country. The statistical analysis of the body and the lung mass of the animals subjected to the single dust and ash insufflation was performed. The applied variants proved the statistically significant differences between the body and the lung mass. The observed differences are connected with the kinds of dust and ash used in the experiment. PMID- 2562680 TI - [About the usefulness of computerized tomography in the diagnosis of intracranial arachnoid cysts]. AB - The authors report 7 cases of intracranial arachnoid cysts diagnosed by the method of computerized tomography in a period of 3 years. In the light of own experiences and after a survey of literature the image of these cysts in CT scans is presented an the problems of differential diagnosis of this disease are discussed. PMID- 2562681 TI - [Cytoenzymatic studies on the influence of soil dust on the cornea and conjunctiva of experimental animals]. AB - The studies of the effect of the soil dust on the cornea and the conjunctiva of the experimental animals have been performed. The rats were divided into 3 experimental groups and 1 control group. The animals were given 3 different, selected at random, kinds of soil dust to the conjunctival sac 3 times daily for 7 days. The results show that the soil dust has the negative effect on the organ of sight. The cornea is particularly sensitive to the action of dust, which evidenced itself by the significant changes in the histochemical reactions. PMID- 2562682 TI - Shock in patients treated in the General Clinic of Internal Diseases, the Medical Academy in Lublin in the years 1979-1988. AB - Shock is, as before, among one of the most serious pathologic states threatening patient's life in spite of considerable progress in its treatment in the recent ten years. The immediate causes of shock may be a decrease of cardiac output, increase of volume of vascular bed or interaction of them both. Decrease of cardiac output may result from defective heart ability to contract or decrease venous blood flow to the heart. Decrease of stroke volume and cardiac output result in defective blood flow through all tissues and lead to metabolic acidosis, whose intensity is decisive for prognosis. PMID- 2562683 TI - Studies on the effect of stress on the estrus cycle in rats. AB - Rats were studied for susceptibility to restraint stress in different phases of the estrus cycle. Susceptibility was measured by the duration of disorders of the estrus cycle. It was found that in proestrus and estrus the disorders of the cycle were most prolonged. During lower sexual activity female rats showed considerable resistance to stress, disorders of the estrus cycle being short lasting. PMID- 2562684 TI - [Some elements of the internal structure of the median nerve in the post-fetal life of man]. AB - The studies were carried out on 240 median nerves taken bilaterally from cadavers of 60 men and 60 women aged from 1 day to 86 years. The number of fascicles in the place where it leaves the brachial plexus, in the upper 1/3 part of the arm, in the lower 1/3 part of the arm and in the cubital area were determined. The magnitude of the surface area of cross-section of the fascicles forming the medical nerve in the lower 1/3 part of the arm were studied. Some of the changes occurring within these elements in the post-fetal life in man were also determined. PMID- 2562685 TI - [Correlation of some elements of the internal structure of the median nerve with age, height and body weight]. AB - The correlation of the number of fascicles of the median nerve with its surface area, as well as the correlation between these elements and age, height and body weight of the examined individuals were studied. A close rectilinear correlation was observed between the size of the area of the cross-section of fascicles of the median nerve and age, height and body weight. The number of fascicles of the medial nerve shows no rectilinear correlation between the size of its surface and age, height and body weight. PMID- 2562686 TI - [Topometry of the bronchial tree in Ceropithecus aethiops with special regard to presence of precordial lobes]. AB - The trachea and bronchus were filled with duracryl solution and on the basis of casts got in this way the length and the diameter of individual lobes of the bronchial tree of 14 male and 16 female monkey have been examined. It was ascertained that almost all measurements were greater in the individuals of female sex. In both individuals among segmental basilar bronchus the greatest diameter had medial and anterior bronchus, both in the right and left lung. PMID- 2562687 TI - [The pulmonary artery size variability during various stages of post-fetal life in man]. AB - Fresh lungs (45 right and 40 left) from dead bodies of 50 (30 male and 20 female) people from 1 to 80 years were examined. The pulmonary arteries were injected for 65% duracryl solution (Spofa-Dental firm) and after maceration the preparations were measured. Pulmonary arteries indices were also calculated. Pulmonary arteries diameter in male measured from 9.0 to 24 mm, average 17.0 mm, in female- from 7.4 to 28 mm, average 18.7 mm. The difference between the right and left pulmonary artery size was statistically significant, both in male and female persons. The correlation indices between size of male right pulmonary artery and age, length and body weight amounted to 0.70, 0.61 and 0.53, while for left pulmonary artery--0.58, 0.55 and 0.49. The same indices of female specimens valued for right pulmonary artery 0.72, 0.58 and 0.51. The size of the left pulmonary artery was correlated only with age (index 0.65). PMID- 2562688 TI - [Tautomerism of 2-hydrazino-4-phenylthiazole<-->4-phenylthiazol-2-one hydrazone. Derivatives of acids. II. (4-phenyl-3-R-thiazol-2-ylidene) and beta-methyl-beta (4-phenylthiazol-2-yl) hydrazides of picolinic, nicotinic, and isonicotinic acid]. PMID- 2562689 TI - Serum iron and total iron binding capacity in larynx cancer patients treated with surgery. AB - A decrease in serum iron and total iron binding capacity (TIBC) has been seen in patients with various malignancies. Similar iron level and TIBC alterations we have found in patients with larynx cancer. Estimation of TIBC reflects the quantity of transferrin which is the iron carrier in serum. The serum iron and transferrin changes cannot be used for diagnostic purposes because they are not specific to malignancies. Such alterations have been also seen in inflammatory and infectious diseases, heart infarct and anaemias. However, it is of interest whether elimination of neoplastic tissue from an organism brings back the decreased serum iron and TIBC to normal values. PMID- 2562690 TI - [Behavioral effects in mice exposed to the prolonged action of cadmium compound and zinc compound]. AB - The aim of the present paper was to determine the effect of cadmium and zinc on the central nervous system in the experimental animals. The animals were given intraperitoneally the solution of cadmium and zinc chloride for 30 days. The behaviour of the mice was being observed in the test of the coordination of movements, the cognitive activity, the spontaneous motility, the motility in the straight rod test and the effect of the examined compounds on the temperature of the animal body was examined. Together with the dose and exposure time increase the cumulative action of cadmium compound was showed, resulting in the inhibiting effect on the central nervous system. The competitive action of zinc compound in relation to the toxic action of cadmium compound was revealed after the administration of the minimal doses. PMID- 2562691 TI - [Toxic-dynamics of lead compound and zinc compound in the evaluation of experimental animal behavior]. AB - The aim of the studies was to determine the effect of lead and zinc compound on the mice behaviour. The animals were given intraperitoneally the solution of lead acetate and zinc chloride for 30 days. The mice were subjected to the tests determining their effects on the coordination of movements, the spontaneous motility, the cognitive motility and the motility in the straight rod test as well as on the body temperature. Due to the prolonged exposition to the action of lead and zinc compounds the inhibiting effect of those substances in the central nervous system in the maximal doses was proved while in the minimal doses the antagonistic action of zinc compound was observed. The nerve conduction disorders in the peripheral and central nervous system after the administration of the maximal doses of lead compound may be due to the inhibiting effect of this compound while the lack of this effect in the minimal doses probably results from the antagonistic action of zinc compound. PMID- 2562692 TI - [Measurements of the arm in inhabitants of the Lublin region]. AB - Anthropological measurements in 1630 inhabitants of the Lublin region were carried out. In all the examined material the length of the right arm in men ranged from 26.5 to 39 cm (mean-32.96) and of the left one--from 26.5 to 38 cm (mean-32.77 cm). The length of the right arm in women ranged from 23 to 36 cm (mean-30.74 cm) and of the left one--from 23 to 36 cm (mean--30.57 cm). The length of the right arm in men was bigger than in women. It was found that both in men and women the right arm is often longer than the left arm. Differences of the length of the arms related to body's side in both sexes were statistically significant. However, no significant differences were found in the length of arms in relation to age. Length index of the arms both in men and women was on the right side 19.7 and on the left side 19.6 and was classified as that of mean arms. PMID- 2562693 TI - [Measurements of the forearm i inhabitants of the Lublin region]. AB - Anthropological measurements in 1150 female and 480 male inhabitants of the Lublin region were carried out. In all the examined material the length of the right forearm in men ranged from 20 to 30.5 cm (mean-25.40), and of the left one- from 20 to 30 cm (mean-25.3); in women the right forearm ranged from 19 to 31 cm (mean-23.50) of length and the left one--from 19 to 33 cm (mean-23.30) of length. The length of the right and left forearms was always bigger in men than in women. Both in men and women the right forearm was found to be longer than the left one and differences between the length of the forearms related to body's side were almost significant in men, and in women--significant. Significant differences were also found in the length of the forearm in relation to age. The length of forearm was decreased in relation to the age in men and in women it was increased. On the basis of arm-forearm index it can be stated that both in men and in women the forearms values were mostly of short class. PMID- 2562694 TI - Measurements of the moving part of upper limb in the inhabitants of the Lublin region. PMID- 2562695 TI - [Clinical freedom]. PMID- 2562696 TI - [The pulsatile administration of LHRH in the differential diagnosis of hypogonadotropic hypogonadism]. AB - The results of the pulsating administration of LHRH (APLHRH) using a portable infusion pumps of national fabrication (TECENSA-AIP-GZ) at doses of 18.2 mcq every 90 min., via subcutanea, in abdominal anterior wall are presented. This was carried out in order to localise the hypogonadotropic hypogonadism. There were statistically significant differences between the group of hypothalamic origin and that of hypophyseal origin after the APLHRH administration. In the first group named, the response patterns of LH and FSH were similar to those obtained in adults, while in the second group there was no response. These differences were not observed at basal conditions nor with acute stimuli of LHRH. We concluded, highlighting the efficacy of this test to diagnose the different types of hypogonadotropic hypogonadism. PMID- 2562697 TI - [Secondary polycythemia and gasometric changes in chronic bronchopneumopathies with respiratory insufficiency]. AB - 42 patients with chronic bronchopneumopathy were divided into 2 groups: one with polyemia (15 patients) and the other without polyemia (27 patients). There was no statistical significant differences in median age, clinical evolution nor basal medium arterial PO2 value (43 +/- 12 and 48 +/- 11 mm Hg); while there was an increase of median basal PCO2 in patients with polyemia (54 +/- 8.4 and 48 +/- 9.3; P < 0.025). There was direct correlation between PCO2 and Hb levels in the whole series (r = 0.42; P < 0.01) and equal correlation in the limit of statistical signification between PCO2 and red cells (r = 0.29; P = 0.05). The correlation between these hematic values and the PO2 values of the whole series was not significant, this taking into account the logical inverse relation between them, because of the important role of hypoxia in developing polyemia. The correlation between PO2 and PCO2 was inverse and significant (r = -0.33; P < 0.05). These findings show the apparent influence of CO2 retention to produce polyemia, apart from the effects of hypoxia, being the opposite of the results in experimental studies which showed a stoppage of the erythropoietin production by CO2 effects. The probable explanation is that patients with more PCO2 had more intense respiratory malfunction than lower PCO2 patients, producing night episodes of more basal hypoxia, this being a major stimulus of polyemia. PMID- 2562698 TI - [Cerebral atrophy in chronic alcoholism]. AB - The brain's morphologic changes induced by chronic abuse of ethanol are studied, using the computerized tomography (CT) of 1005 clinical records reviewed during 1982-1987 at "Hospital Universitario de Granada", only 33 were selected as patients suffering from chronic alcoholism, CTs having been performed. The results were compared with 33 CTs of non-drinking patients. The ventricular index (VI), the evans' index (EI), the transversal diameter of third ventricle (TD) and cerebral sulcus evaluation were the values analysed to estimate the degree of cerebral atrophy. It was proven that a significant decrease of VI, an increase of EI, TD and cerebral sulcus in chronic alcoholic patients exists compared to non drinkers. 70% of the alcoholic patients had cerebral atrophy. The conclusion of this study is that chronic alcoholism is an important causal factors of the cerebral atrophy process, regardless of age, alcohol intake quantity and/or the existence of other associate pathology. PMID- 2562700 TI - [The usefulness of cholesterol determination in the etiology of pleural effusion]. AB - 95 pleural effusions of different etiology were studied. 25 were transudate, 22 neoplastic, 28 caused by tuberculosis, and 20 miscellaneous. The Light parameters (pleural protein/plasma) (Prot LP/P), Pleural LDH (LDH LP) and LDH pleural/plasma quotient (LDH LP/P) were determined and compared with the efficacy of pleural cholesterol (Chol LP) and pleural cholesterol/plasma quotient (Chol LP/P) to differentiate the transudate from pleural exudate. With Prot LP/P quotient the sensitivity was of 85% and the specificity 91.6%; with LDH LP the sensitivity was of 74.6% and the specificity 97.6%. With the LDH LP/P quotient the sensitivity was of 89% and the specificity 87.8%. The sensitivity of each parameter was similar to that obtained by Light but less specific. All the transudates had levels of chol LP lower than 55 mq/dl (100% specificity) with a sensitivity of 84%. The Chol LP/P quotient had a limit of 0.3, this being its sensitivity of 91.3% and 93.1% its specificity. The presence of both parameters were necessary to the classification and the sensitivity obtained being of 80.3% and the specificity 100%. We concluded that the determinations of Chol LP and the Chol LP/P quotient are of great advantage in differentiating the pleural transudate from pleural exudate and the predictive value is, at least, similar to that obtained by Light. PMID- 2562699 TI - [IgM, IgG and IgA rheumatoid factors in rheumatoid arthritis]. AB - The IgM, IgG and IgA rheumatoid factors (RF) were studied by ELISA in the serum of 122 patients with rheumatoid arthritis (RA) associating the Waaler-Rose test results, with the clinical and radiological aspects of the disease. 75 patients (61%) had RF IgM positive according to the Waaler-Rose test while the ELISA showed positive in 104 (85%). The RF (IgG was positive in 64 cases (52%) and RF IgA in 82 (67%). The levels of RF IgA were correlated to RF IgM levels, determined by the Waaler-Rose test (p < 0.01) and ELISA (p < 0.001), while RF IgG levels were not correlated to RF IgM or IgA. There was a significant correlation between RF IgA titles and Lansbury's index (p < 0.01), and between RF IgG and sedimentation rate (p < 0.01). In patients with extraarticular disease high levels of RF have been observed, especially RF IgM and IgA. We concluded that the ELISA technique is the preferred method to measure the RF. PMID- 2562701 TI - [Pulmonary infiltrates, eosinophilia and polyneuritis]. AB - A case of chronic eosinophilic pneumonia which, during the evolution, developed a very rare motor-sensitive polyneuritis of the lower limbs, is presented. We comment on the results of the electromyography, the neuromuscular biopsy and show the differential diagnosis with other diseases with lung infiltrates, peripheric eosinophilia and polyneuritis, such as hypereosinophilic syndrome, allergic angiitis and granulomatosis of Churg and Strauss. PMID- 2562702 TI - [Hepatotoxicity induced by cyanamide. A review of the anatomicopathological changes apropos a case]. AB - A case of toxic liver disease induced by cyanamide in a patient in treatment with this drug during 18 months, is presented. We reviewed the different liver cell alterations produced by a long-term treatment with cyanamide, which produces fibrosis and portal inflammation, as well as polished cells with different characteristic features. These alterations forced the establishing of close controls of patients in treatment with this type of anti-alcoholism drug, as well as the reduction of the duration of therapy, this questioning the efficacy of the treatment of chronic alcoholism with this aversive drug. PMID- 2562703 TI - [Atypical relapsing polychondritis]. AB - Relapsing polychondritis is a disease of unknown etiology whose main characteristic is the chronic inflammation and destruction of the different cartilaginous structures of the body. A rare case is presented, the rareness being the fact that ear cartilage was not affected and that the destructive arthropathy was similar to that produced by rheumatoid arthritis. PMID- 2562704 TI - [Acute interstitial secondary to tricuspid endocarditis caused by Staphylococcus aureus]. AB - The kidney diseases in patients with bacterial endocarditis and intravenous drug addicts (IVDA) tend to be of glomerular origin. Interstitial nephritis has been related to drug toxicity and only occasionally has it been described in other associations. We describe a 27-year-old patient IVDA with tricuspid endocarditis caused by S. Aureus whose first manifestations was acute renal failure. The renal biopsy showed an interstitial nephritis. It was treated with antibiotic and hemodialysis, obtaining the cure and normal levels of plasmatic creatinine. PMID- 2562705 TI - [The current possibilities in the diagnosis of hypogonadotropic hypogonadism]. AB - The diagnosis of hypogonadotropic hypogonadism must be clinically suspected. They produce differential diagnosis problems. The possibilities of diagnosis are reviewed (static hormonal determination, dynamic test-clomiphene test, LHRH acute test, etc.), highlighting the pulsating administration of LHRH which, together with endogenous pulsating of LHRH study (more difficult), are the most useful procedures in differentiating the level of the lesion of hypogonadotropic hypogonadism, as well as differentiating the puberty delayed cases. PMID- 2562706 TI - [Salmonella osteomyelitis in an AIDS patient]. PMID- 2562707 TI - [Primary empty sella turcica and spontaneous cerebrospinal rhinorrhea. A case report]. PMID- 2562708 TI - [The efficacy of oral ciprofloxacin in the treatment of Proteus mirabilis infections of the lower respiratory tract]. PMID- 2562709 TI - [Lumbar tumefaction as the form of presentation of tuberculous spondylodiscitis]. PMID- 2562710 TI - [Pulmonary thromboembolism as the first manifestation of systemic lupus erythematosus]. PMID- 2562711 TI - [Paroxysmal arterial hypertension associated with a "nonfunctioning" adrenal adenoma]. PMID- 2562712 TI - [Auricular fibrillation and the alcohol withdrawal syndrome: a new case]. PMID- 2562713 TI - [Chromosome X-linked recessive bulbospinal neuronopathy (Kennedy's syndrome)]. PMID- 2562714 TI - [Primary familial spontaneous pneumothorax]. PMID- 2562715 TI - [Arterial hypertension in the elderly: a therapeutic problem]. PMID- 2562716 TI - [Bullous diseases and neoplasms]. PMID- 2562717 TI - [Erythrocyte morphometry in alcoholism]. AB - The aim of this study is to evaluate the erythrocyte morphology (area, perimeter, shape) and its correlation with high alcohol intake and the conventional blood test (MCV, GOT/GPT, and activated PTT). 60 persons were studied (20 non-drinkers, 20 chronic alcoholics and 20 with hepatic cirrhosis). The erythrocyte area and perimeter was significantly higher, in patients other than the group of non drinkers. This is directly related to the alcohol intake and liver damage. We believe, that the study of the morphology of the erythrocyte is of interest as a "biological marker" related to the grade of alcoholism. PMID- 2562718 TI - [The evaluation of the diagnostic usefulness of the clinical picture, electrocardiography and enzymes in the initial presentation of an acute myocardial infarct]. AB - Medical data of 181 patients, affected by confirmed acute myocardial infarction, were reviewed to evaluate the initial clinical, electrographic and radiological data, and the first CPK test. Eleven patients went to the emergency department without chest pain (6.1%). The percentage of patients with oppressive chest pain was 73%, being of thoracic localization in 86.4%, 20%. Related to effort; 47.1% referred; and 92.9% of more than 30 mins. duration. 35.3% had normal blood pressure. Only 5 patients (2.8%) had normal EKG, the S-T segment elevation being the most frequent alteration (71.3%). 89.5% of patients had sinusal rhythms; 20.4% were bradycardic. 58.6% had normal levels of CPK at the first test. We concluded that EKG is the most important test in diagnosing acute myocardial infarction; it being very rarely normal. The clinical data are coadjuvant, while isolated initial CPK had not value. PMID- 2562719 TI - [Follicular thyroid cancer: the prognostic factors]. AB - 30 patients affected by follicular carcinoma of the thyroid gland (well differentiated, poorly differentiated and Hurthle's cell) are studied. We analyse the factors which affect the mortality; the percentage was of 63% during 3 years follow-up of metastatic patients and was null in non-metastatic patients (P < 0.0001); without significant differences between sex, age and histological type. Patients without metastasis at diagnosis had increased TG in 60% of the Hurthle's cell carcinomas and in 30%. Of the follicular carcinomas; poorly differentiated and not detected in 100% of the well differentiated follicular carcinoma, after a median follow-up of 7 years. This suggests that prognosis might be different depending of histological type. PMID- 2562720 TI - [The incidence of consumption coagulopathy in liver cirrhosis]. AB - DIC in patients affected by cirrhosis, accompanied by portal hypertension and splenomegaly, has been suspected in the past. The main aim of this study is to ascertain the incidence of this phenomenon. We carried out coagulation and fibrinolytic tests in 113 cirrhotic patients and 20 healthy control persons. We found chronic consumption coagulopathy at analysis level in 28 cases (24.8%) with a decrease of fibrinogen, factor V, kallikrein, platelets, prothrombin complex activity, increase of PDF, partial thromboplastic time and euglobulin lysis. 25 cases had active cirrhosis, with ascites, variceal bleeding and/or hepatic encephalopathy; 3 were non-active cirrhosis. Only 7 patients had clinical DIC. We observed that coagulation disorders increased with more active cirrhosis. PMID- 2562721 TI - [A morbidity study in a general internal medicine service in a third-level hospital]. AB - This study carried out at a type "C" hospital, analyses the actual pathology of 1,052 patients attended to at the internal medicine department during a period of one year. The sex distribution did not show any differences. The median age (64 years) was significantly superior in women. The more frequent diseases were from group VII (cardiovascular: 512 cases) and group VIII (respiratory: 471 cases) according to the 9th edition of the who international diseases classification. The most frequent causes for admission were: respiratory infection (19.5%), cardiac insufficiency (13.8%) and CVA (10.6%). The most frequent baseline diseases were cardiomyopathy (20.4%), chronic obstructive airways syndrome (16%), malignant neoplasia (8.5%) and hepatopathy (7.6%). The risk factors and toxic habits observed were: Chronic bronchitis (19.6%), blood hypertension (15.5%), diabetes (13.5%) and high alcohol intake (10%). PMID- 2562723 TI - [The superior vena cava syndrome in retrosternal goiter]. AB - A case of superior vena cava syndrome in an 82-year-old woman who had been suffering from multinodular hyperfunctioning goiter for a long time is presented. The goiter was partially intrathoracic, benign, but affecting the superior mediastinum. The patient decided not to be operated on because of personal fears and the slow growth produced developments of collateral prethoracic circulation. The etiology of superior vena cava syndrome is of malignant tumor origin in 95% of cases. This case is presented because of its rareness, and the literature is reviewed. PMID- 2562722 TI - [Endobronchial foreign bodies in adults. A report of 3 cases with a prolonged evolution]. AB - 3 cases of endobronchial foreign body (EFB) in adult patients which main feature was a delay in diagnosis, are presented. The aspiratory accident was not well evaluated by the patients. There were several complications (hemoptysis, pneumonia, and empyema) during the interval from the aspiration to the extraction, these being the reasons for inducing diagnosis. The mechanism of aspiration and the treatment is discussed. PMID- 2562724 TI - [Autoimmune hemolytic anemia preceding by 6 years an adenocarcinoma of the gallbladder]. AB - A case of a 59-year-old female suffering from immunohemolytic anemia, coombs negative, is presented. Six years after the diagnosis, this patient developed an adenocarcinoma of the gallbladder, with invasion of lymph nodes, liver and peritoneum; at this moment the coombs, test proving positive. The possible pathogenetic mechanisms are discussed, despite the rareness of this delayed association which is probably the first described in literature. PMID- 2562725 TI - [Lead poisoning due to drug addiction: a new source of poisoning with clinical interest and important epidemiological consequences]. AB - Lead poisoning has accompanied the human being throughout history. Owing to the increasing levels of safety at work, the incidence of occupational poisoning has decreased and new forms of non-occupational poisoning have emerged. We present 3 cases of drug addicts, with lead poisoning, as a result of using adulterated drugs. One of them was an intravenous drug addict who had abdominal pain and anemia. The other 2 inhaled heroin, one being slightly anemic and the other without symptoms and with normal hemoglobin levels. The drug adulterated with lead had not been previously recognized as a source of lead poisoning, being likely to cause serious epidemiological effects. PMID- 2562726 TI - [Carbon tetrachloride poisoning: a report of 3 cases]. AB - Carbon tetrachloride is a toxic solvent easily obtained in our country. 3 cases of poisoning by accidental inhalation at place of work. The main clinical manifestations were acute renal failure and toxic hepatopathy. All patients, had a good evolution with dialysis therapy after a few weeks. We comment on some possible additional therapies to be used in treating patients with severe poisoning due to carbon tetrachloride. PMID- 2562727 TI - [Bacterial adherence as a virulence factor in sepsis]. AB - In the present revision we are doing a study on the bacterial adherence in the develop of sepsis. The adherence is a important phenomenon in the initial stages of infections, but when the microorganism go in the blood her presence assure the posterior union to the phagocytic cells. The administration of sublethal concentrations of antibiotics suppress the formation and expression of bacterial adhesins. PMID- 2562728 TI - [Primary hypothyroidism followed by hyperthyroidism]. PMID- 2562729 TI - [Devic's syndrome during secondary syphilis]. PMID- 2562730 TI - [A case of eosinophilic fasciitis with myositis]. PMID- 2562731 TI - [The treatment of hypertension with captopril in the elderly patient with multiple pathologies]. PMID- 2562732 TI - [Haemophilus influenzae pneumonia in patients with the human immunodeficiency virus]. PMID- 2562733 TI - [Hepatitis delta virus infection in asymptomatic carriers of the hepatitis B virus]. PMID- 2562734 TI - [Shunt-related nephritis]. PMID- 2562735 TI - [The tobacco habit, hypertension and vascular risk; a controversial relationship]. PMID- 2562736 TI - [Mediterranean boutonneuse fever and pneumonia]. PMID- 2562737 TI - Occupational exposure to anaesthetics: liver injury, microsomal enzyme induction and preventive aspects. AB - Apart from a risk excess of liver disease among operating theatre personnel and of spontaneous abortion in women exposed during pregnancy, no definitive conclusion has been drawn as regards health impairment among anaesthesiology staff. Hepatotoxicity has been detected in experimental and epidemiological studies, suggesting a close relationship between liver disease and anaesthetics, particularly halogenated hydrocarbons (halothane and isoflurane) and nitrous oxide. The liver microsomal enzyme system has received particular attention in order to clarify the mechanism involved in anaesthetics hepatotoxicity and an increased microsomal enzyme activity has been observed in experimental conditions and in humans (both patients treated with anaesthetic mixture and anaesthesiology staff). The prevention of adverse health effect among workers of anaesthesiology staff, is based on (i) the use of less toxic anaesthetics and (ii) the control of working conditions of staff. Attention is focused on biological monitoring by means of indicators, which reflect not only the environmental pollution but can be regarded as liver response of anaesthetics exposed workers. This inductive effect is commonly considered the earliest sign caused by exposure to several chemicals and may be evaluated by means of biomarkers, among which the measure of urinary D-glucaric acid excretion is a well established non invasive tool. PMID- 2562738 TI - [Acute accidental poisoning by mercury vapors in the home environment]. AB - The authors report 2 cases of acute mercury intoxication due to accidental breakage of barometer on to a lit gas ring. Within 24-48 hours the subjects developed neurological, gastrointestinal and dermatological symptoms. A 24-hour urine sample contained 600 and 400 micrograms of mercury per liter respectively (reference value 0.1-6.9 micrograms/L); blood concentration of mercury was 130 and 100 micrograms per liter (reference value 1.7-9.9 micrograms/L). The patients were treated with penicillamine and daily excretion of mercury was monitored. The residual sources of pollution in the kitchen were identified and bonificated. PMID- 2562739 TI - [An annual decrement in the spirometric parameters of those employed in producing refractory materials. A longitudinal study]. AB - Results of a longitudinal study on the yearly decline of two pulmonary parameters are reported. All subjects were workers in a refractory material factory and exposed to dust with low silica content. The study brought about an annual measurement of two spirometric parameters (VC and FEV1) during a 13 year period of follow up on 113 subjects. 56% and 57.5% of them showed annual decreases of VC and FEV1 statistically significant. The trend of the two functional parameters over time was studied both in the whole group of workers and in subgroups obtained from occupational exposure levels, smoking habit, age. The most affected volumetric index was the VC. The 3 risk indicators considered (occupational exposure, smoking habit and age) all negatively influence the VC decrease; however, the effect is statistically significant only for smoking habit. The FEV1 changes with reference to the above mentioned risk indicators are less clear. From the whole group a number of 15 losers were selected. PMID- 2562740 TI - [Occupational asthma]. AB - Bronchial asthma has been defined as a state of airway obstruction of variable degree correlated casually with exposure to volatile dusts, gases, vapours or fumes in the working environment. In this review we considered the causes of occupational asthma, the diagnosis, the pattern of asthmatic reactions and their mechanism, the predisposing factors, the epidemiology, the prognosis and finally the cure and prevention (review article, 165 references). PMID- 2562741 TI - [The determination of iron, lead, copper and zinc in the hair of children between 22 and 85 months old]. AB - In this study the levels of Cu, Fe, Pb and Zn, determined in hair of healthy children (25-85 months), living at Araraquara (Sao Paulo, Brasil) are reported. Analytical determinations were carried out by using atomic absorption spectrometry employing an air acetylene flame. The results obtained are discussed with regard to effect of sex and age, as well compared with others analogous studies. PMID- 2562742 TI - Erythrocytes parameters due to aging, smoking, alcohol consumption and occupational activity in a working population of petrochemical industry. The Pavia Study. AB - The main purpose of the study was to assess the red blood cell disorders in works employed in the petrochemical industry in the Lombardia region (ENI). In the analysis of the data the confounding factors such as age, smoking habit, alcohol consumption, nutritional status based on body mass index have also been taken into consideration. In total, material consisted of 1175 male workers in whom red blood cell examination of the peripheral fasting blood was carried out. The results of the study showed that clinical anemia has been found in 1.7% of subjects examined. The distribution of anemia cases was not related to job category or physical occupational activity, neither to age, smoking or alcohol consumption. However, the results of the study suggest that aging processes are associated with weakening of hemopoietic system which affect in a great extent hemoglobin production. The effect of smoking on hemopoietic system appeared to have different features from that attributed to aging. Smoking increased hemoglobin level and hematocrit significantly in comparison to nonsmokers, but had no effect on the number of erythrocytes. It was concluded that adaptation to carbon monoxide inhaled with cigarette smoke is reflected by an increased red cell mass and hemoglobin. Occupational factors measured by the type of job (manual vs nonmanual) appeared to have no harmful effect on the hemopoietic system, however, respondents who reported working physically in an intensive way, showed significantly lower mean corpuscular concentration hemoglobin level than those having a sedentary job or engaged in very small physical activity at work. PMID- 2562743 TI - [Allergic contact dermatitis and oculorhinitis due to tulips]. AB - A case of occupational allergic contact dermatitis with conjunctivitis and rhinitis due to tulips bulbs is described. The patient, aged 23, was employed in a florist storehouse. The dermatitis, affecting his hands, face, armpits, knees, elbows and ears, was accompanied by a rhinoconjunctivitis. Patch tests made with a tulip thin slice outcome positive results. The appearance, the way of arising and the results of the patch tests suggest the existence of an immediate type immunologic reaction (type I or IgE-dependent hypersensibility). PMID- 2562744 TI - [Electromyography of those exposed to vibration]. AB - Electrophysiological findings in the median and ulnar nerves of a group of ten workers who operate hand-held vibrating power tools are described. Motor conduction velocities, motor latencies and amplitude of evoked compound muscle action potentials were measured. We have observed statistically significant changes in the distal latencies and in the amplitude of evoked compound muscle action potentials of median nerves in all the subjects examined, whereas the motor conduction velocities were normal in both median and ulnar nerve. These findings are compatible with functional changes without a clinically manifest neuropathy and they are suggestive of a compression damage such as carpal tunnel syndrome: the possible pathogenic mechanism of these damages are discussed. It's necessary to emphasize the importance of further studies in order to estimate the type and entity of the exposure to vibrations to show a cause-effect relationship. PMID- 2562745 TI - Methyl ethyl ketone (MEK) in urine as biological index of exposure. AB - Fifteen human volunteers were exposed to methyl ethyl ketone (MEK) vapor at 11.9 621.8 mg/m3 for a period of 2 to 4 hours at rest (ten cases) and during light physical exercise (five cases). Subsequently 78 workers occupationally exposed to MEK in a manufacture of leather suitcases (median value: 75.5 mg/m3; geometrical standard deviation: 3.12 mg/m3; range: 6-790) were studied. The analyses were performed by means of a Gas Chromatograph (GC) Hewlett-Packard 5880 A connected with a Mass Selective Detector (MSD). The relative uptake (R) of MEK was about 0.54 (standard deviation: 0.05) and it keeps practically constant either at rest or during light effort (V < 30 L/min). A linear relationship existed in the experimentally exposed subjects between urinary concentration (Cu) and amount of MEK absorbed (U) (Cu = 3.05 x U-162.1; r = 0.95; n = 15) (Cu = micrograms/L; U = mg). Both in the experimentally exposed subjects and in the occupationally exposed workers, the urinary concentration of MEK shoved a linear relationship to the corresponding environmental time-weighted average concentration (CI). The correlation coefficients (r) were 0.93 in occupationally exposed subjects (regression equation: Cu = 0.004 x CI + 0.118; n = 78); Cu = mg/L; CI = mg/m3) and more than 0.93 in experimentally exposed groups. The findings indicate that the urinary concentration of MEK can be used as an appropriate biological exposure indicator.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562746 TI - [Qualitative and quantitative changes in the polycyclic aromatic hydrocarbons in cutting oils after their use]. AB - The quantitative and qualitative variations in the mixture of polycyclic aromatic hydrocarbons (PAH) present in different samples of a cutting oil, unused and after 3,6 and 9 months of use, were evaluated by means of gas-chromatography mass spectrometry. Nine of the identified hydrocarbons (phenanthrene, anthracene, fluoranthene, pyrene, benzo-a-anthracene, chrysene, triphenylene, benzo-a-pyrene, benzo-e-pyrene and perylene) were studied. The total PAH concentration increased from 45 (first sample) to 915 ng/gr of oil, even if a different behaviour for various hydrocarbons was shown. In fact some of them, such as phenanthrene and anthracene increased with use, some, such as fluoranthene and pyrene decreased and the other did not exhibit a regular trend. In the light of results, the influence of variations of the PAH mixture in oil on the PAH air concentration and on the preventive measures to be adopted, is discussed. PMID- 2562747 TI - [Asthenopia and objective ophthalmological findings in a population of 2058 VDT operators in Lombardy]. AB - A large population of 2058 VDT operators from all compartments in Lombardia of a large national company was studied. An "ergophthalmological" questionnaire was administered to all subjects, followed by an ophthalmological-orthoptic examination. Ametropias showed a prevalence of 51.9%; the most common refractive defect was myopia followed by astigmatism and hyperopia. Heterophorias with bad or mediocre compensation had a prevalence of 4.1 and 11% respectively. The study of symptoms showed a frequency of 23.5% of intense asthenopic complaints and 21.1% of less severe and less frequent symptomatology. Asthenopia was significantly correlated (chi square) with sex, VDT exposure, refraction, presbyopia, and decompensated heterophorias. PMID- 2562748 TI - Coronary heart disease risk score and ECG abnormalities in apparently healthy workers from petrochemical industry. The Pavia Study. AB - The purpose of the study undertaken was to define the vulnerable fraction of the industrial population in northern Italy using the CHD risk score profile. In addition, ECG examinations were carried out in order to see whether there is a correlation between cardiovascular risk profile and the frequency of ECG abnormalities. The study covered in total 1443 male workers employed in various petrochemical plants. The calculation of the CHD risk score was based on age, relative weight, serum cholesterol, systolic blood pressure, smoking habit, physical activity at work and at home, and heredity factors. The study demonstrated that on an average 47.3% of male workers in the petrochemical industry have high or very high risk of cardiovascular disease and this draws the attention to the urgent need to develop the better and more effective primary preventive strategy against cardiovascular diseases, specially among manual workers. The study disclosed also that the persons with elevated risk score displayed more frequently ECG repolarization disorders. PMID- 2562750 TI - [The evaluation of the individual thermal load in work activity with uneven energy expenditure]. AB - It is often very difficult to quantify the potential thermal stress in jobs characterized by sudden variations of energy expenditure and thermal overload for time periods of short duration. Heart rate varies rapidly with the changes of energy expenditure and thermal load and the application of the mathematical model, proposed by Givoni and Goldmann for prediction of heart rate response to work in hot environments, to values of the physiological parameter, continuously measured during the periods of work and rest, allows the calculation of the two components of the HSI stress index (Er, Emax) from which to derive safe times of exposure. In this study practical applications of the theoretical method are verified in the control of the potential thermal stress of workers engaged in the carbon coke production. In that job the variability of thermal conditions make unfeasible a complete and whole evaluation of thermal overload only based on the measure of the microclimatic environmental parameters. PMID- 2562749 TI - [The phagocytic activity of the neutrophilic granulocytes via chemoluminescence and occupational exposure to lead]. AB - The lead interference on the immunological activity is the aim of this survey. Therefore, it was examined the phagocytic function of the PMN cells on a group of 20 subjects occupationally exposed to the toxic and on 20 other assigned to the control group using the chemoluminescent assay. In both groups it was determined the biologicals parameters of exposure to the toxic. It was evident a statistically significant decrease of the average values in the exposed group compared to the control group, even if it wasn't directly correlated with the exposure biological indicators. PMID- 2562751 TI - [A ergonomic chair on an oscillating pivot with an electrical motor for the prevention of occupational lumbar pathology due to the seated posture]. AB - It is described an ergonomic chair of new conception, because it is not a static one, but slowly swinging in all planes, with a motion activated by an electrical engine. This chair is patented. The clinical experimentation on cases of "minor" lumbar pathology, as it is found more and more frequently in prolonged seated working situations, confirm the benefit of this instrument, already tested in cases of manifest lumbar pathology. PMID- 2562752 TI - [Recent findings on the epidemiological and experimental aspects of the measurement of serum bile acids in occupational exposure to xenobiotics]. AB - The functional activity of the liver and the variety of its responses to injury makes the choice of appropriate tests a difficult task. However, because of the highly efficient uptake of bile acids by the normal hepatocyte, the determination of serum bile acids (SBA) concentration has been proposed as a test to detect early changes of liver function not associated to cytotoxicity. Several studies of biomonitoring subjects occupationally exposed to hepatotoxic substances have been carried out by evaluating total SBA or primary SBA, as indicators of early liver dysfunction. Even though these studies are not completely comparable because of the different protocols adopted, most of them reveal a significant increase of SBA concentrations among the exposed subjects in respect of unexposed controls. Furthermore, higher prevalences of subjects exposed to organic solvents mixture with abnormal SBA concentrations in respect of controls have been observed. Increased serum bile acids concentrations among the subjects exposed to a variety of xenobiotics have been explained as a change in hepatocyte function, particularly in one of the steps involved in bile acids transport. However, the lack of any relationship between the indices of doses and SBA concentrations remains an important point to clarify. This fact could be interpreted as a consequence of the delayed biological effect responsible for increased serum bile acids during the exposure. As regards the nature of the mechanisms involved in the increase of SBA concentrations, recent observations pointed out that some chlorinated aliphatics were able to inhibit cell membrane ATPases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562753 TI - [Fatigue during muscle work at variable traction angles]. AB - Using computerized surface EMG signal analysis, authors point out that muscle fatigue onset depends not only by exercise frequency and external load but also by mechanical arrangement of muscle insertions and this onset is faster if contraction is concentric. Therefore selective muscle training programs must contemplate this phenomenon. PMID- 2562754 TI - [The prevention of occupational exposure to anesthetics: the standards aspects]. AB - Prevention of damage of occupational exposure to anaesthetic gases and vapors consists in limitation of pollution and in biological and environmental monitoring. Reduction of polluting concentrations can be obtained only by using both ventilation of the operating room and active scavenging; proper behaviour in accordance with prevention rules and psychologic involvement of personnel is also needed. Biological and environmental measurements are useful in order to evaluate the efficiency of prevention and to early detected overexposures. Acts and regulations in force at present in Italy and technical specifications concerning this problem are than reviewed. The author notices that in many cases, standards dealing with requirements to be satisfied should be established before performance standards; at any rate, regulations should also be more compulsive. PMID- 2562755 TI - Production of root hair deformation factors by Rhizobium meliloti nodulation genes in Escherichia coli: HsnD (NodH) is involved in the plant host-specific modification of the NodABC factor. AB - The role of the hsnD (nodH) gene in the determination of the host-specific nodulation ability of Rhizobium meliloti was studied by expressing the common nodulation genes (nodABC) with or without the hsnD gene in Escherichia coli and testing for biological activity on various leguminous plants. In this way, four categories of plants were established. Upon infection with E. coli carrying the nodABC construct, root hair deformation (Had) was detected on clovers while the hsnD gene was additionally needed for the elicitation of the same response on alfalfa and sweet clover. A weak root hair deformation was seen on siratro by inoculation with E. coli harbouring the nodABC genes and was highly increased when hsnD was also introduced. Cowpea and Desmodium did not respond to any of the E. coli strains constructed. Exudates or cytosolic fractions of the respective E. coli derivatives elicited the same root hair deformation as the intact bacteria. These data indicate that not only the nodABC gene products but also the hsnD product are involved in the synthesis of Had factors. Subclones expressing only the nodA, nodB, or nodC genes or the same genes in pairs (nodAB, nodBC, nodAC) did not provide a compound with activity comparable to the NodABC factor, suggesting that all three genes are required for the production of the Had factor which is active on clover. Coinoculation of alfalfa plants with two strains of E. coli, one carrying the nodABC genes and the other expressing only hsnD, or combining exudates or cytosolic fractions from these strains did not result in root hair deformation on alfalfa. These data indicate that the HsnD protein itself or its product is not an additional alfalfa-specific extracellular signal but more likely is enzymatically involved in the modification of the basic compound determined by the nodABC genes. PMID- 2562756 TI - The small subunit of ribulose-1,5-bisphosphate carboxylase is plastid-encoded in the chlorophyll c-containing alga Cryptomonas phi. AB - The gene for the small subunit of ribulose-1,5-bisphosphate carboxylase (Rubisco) is located in the large single-copy region of the plastid genome of the chlorophyll c-containing alga Cryptomonas phi. The coding sequence is 417 base pairs long, encoding a protein of 139 amino acids, considerably longer than most other small subunit proteins. It is found 83 base pairs downstream from the gene for the large subunit and is cotranscribed with it. An 18 base pair perfect inverted repeat is located 8 base pairs beyond the termination codon. Sequence analysis shows the gene to be more closely related to cyanobacterial and cyanelle small-subunit genes than to those of green algae or land plants. This is the first reported sequence of a Rubisco small-subunit gene which is plastid-encoded and it exhibits a number of unique features. The derived amino acid sequence shows extensive similarity to a partial amino acid sequence from a brown alga, indicating that this gene will be of major interest as a probe for the small subunit genes in other algae and for determining possible evolutionary ancestors of algal plastids. PMID- 2562757 TI - Transient gene expression in aleurone protoplasts isolated from developing caryopses of barley and wheat. AB - Methods have been developed for the isolation of aleurone protoplasts from developing caryopses of Hordeum vulgare and Triticum aestivum in order to study transient expression of introduced genes. Chimaeric gene constructs were introduced into aleurone protoplasts by polyethylene glycol (PEG). Transient expression directed by the 35S promoter from cauliflower mosaic virus (CaMV) of the reporter gene encoding chloramphenicol acetyl transferase (CAT) was detected in aleurone protoplasts from developing barley and wheat grains. Using a similar construct, CAT activity increased when the alcohol dehydrogenase intron 1 fragment from maize was ligated between the 35S promoter and the CAT coding region. The demonstration of transient expression in protoplasts from developing aleurone layers indicates that they may be useful for investigating tissue and developmental control of genes coding for cereal seed proteins. PMID- 2562758 TI - Purification of (1-->3)-beta-glucan endohydrolase isoenzyme II from germinated barley and determination of its primary structure from a cDNA clone. AB - A (1-->3)-beta-D-glucan 3-glucanohydrolase (EC 3.2.1.39) of apparent M(r) 32,000, designated GII, has been purified from germinated barley grain and characterized. The isoenzyme is resolved from a previously purified isoenzyme (GI) on the basis of differences in their isoelectric points; (1-->3)-beta-glucanases GI and GII have pI values of 8.6 and > or = 10.0, respectively. Comparison of the sequences of their 40 NH2-terminal amino acids reveals 68% positional identity. A 1265 nucleotide pair cDNA encoding (1-->3)-beta-glucanase isoenzyme GII has been isolated from a library prepared with mRNA of 2-day germinated barley scutella. Nucleotide sequence analysis of the cDNA has enabled the complete primary structure of the 306 amino acid (1-->3)-beta-glucanase to be deduced, together with that of a putative NH2-terminal signal peptide of 28 amino acid residues. The (1-->3)-beta-glucanase cDNA is characterized by a high (G+C) content, which reflects a strong bias for the use of G or C in the wobble base position of codons. The amino acid sequence of the (1-->3)-beta-glucanase shows highly conserved internal domains and 52% overall positional identity with barley (1- >3, 1-->4)-beta-glucanase isoenzyme EII, an enzyme of related but quite distinct substrate specificity. Thus, the (1-->3)-beta-glucanases, which may provide a degree of protection against microbial invasion of germinated barley grain through their ability to degrade fungal cell wall polysaccharides, appear to share a common evolutionary origin with the (1-->3, 1-->4)-beta-glucanases, which function to depolymerize endosperm cell walls in the germinated grain. PMID- 2562759 TI - Induction and growth properties of carrot roots with different complements of Agrobacterium rhizogenes T-DNA. AB - Single and multiple infections of carrot discs were carried out with Agrobacterium strains harbouring different segments of pRi1855 TL-DNA cloned in the binary vector Bin 19 and with a strain carrying the TR-DNA from the same Ri plasmid. Roots induced by the various co-inoculations were cultured and their growth patterns were followed. Abundant roots could be induced by TL-DNA rol genes A, B and C as a single insert (rolA + B + C) and by rolB alone provided an extended segment beyond its 5' non-coding region was included in the construction. A depression of rooting capability was caused by the inclusion of rolC together with rolB (rolB + C). In all cases co-inoculation with the Agrobacterium carrying TR-DNA-borne auxin genes was necessary for root induction since none of the rol constructions was in itself capable of eliciting any response; an exceeding majority of these roots were however shown to contain rol genes but no TR-DNA. Rooting was also elicited if rol constructions were co inoculated with a strain carrying TL-DNA genes 13 and 14 (ORF13 + 14) instead of the TR-DNA strain. These roots were shown to contain both rol genes and ORF13 + 14. Striking differences in growth properties were shown by roots containing different complements of TL-DNA genes. Typical hairy root traits, high growth rate, branching and, most noticeably, absence of geotropism, were shown by roots containing rolB alone, while roots with rolA + B + C were geotropic as normal carrot roots. Hairy root traits were conferred to rolA + B + C roots by the concomitant presence of ORF13 + 14 and by the addition of auxin to the culture medium. A model is presented which attempts to rationalize the growth patterns by assigning interplaying roles to the various TL-DNA genes involved. PMID- 2562760 TI - Rice alcohol dehydrogenase genes: anaerobic induction, organ specific expression and characterization of cDNA clones. AB - Anaerobiosis rapidly induces alcohol dehydrogenase (ADH), an enzyme of the fermentation pathway, in different parts of rice seedlings. After initiation of anaerobiosis, the activity of the enzyme increases linearly for 3 days or more. The ADH activity is anaerobically inducible even in mature rice leaves in contrast to maize which shows no induction in mature leaves. Rice ADH activity can also be induced by an auxin analog, 2,4-dichlorophenoxyacetic acid, under aerobic conditions. The experimental results show that anaerobiosis increases the ADH mRNA level, indicating that the ADH enzyme is regulated at the transcriptional level. Starch gel electrophoresis of a protein extract from rice shows 3 distinct forms of ADH. The amounts of the 3 forms vary with the organ, suggesting that the expression of ADH genes is organ-specific. Sequencing data show that the two different cloned cDNA copies of ADH mRNAs are derived from two different genes. PMID- 2562761 TI - Cloning and expression of the chloroplast-encoded rbcL and rbcS genes from the marine diatom Cylindrotheca sp. strain N1. AB - Both the rbcL and rbcS genes, encoding the large and small subunits, respectively, of ribulose 1,5-bisphosphate carboxylase/oxygenase, have been found to be encoded by chloroplast DNA in the marine diatom Cylindrotheca sp. N1. The rbcS gene in this diatom was found to be adjacent to the rbcL gene by a combination of: (i) Southern-blotting analyses, using heterologous probes; (ii) examination of recombinant proteins synthesized in Escherichia coli, directed by cloned rbcL/rbcS genes; and (iii) synthesis of enzymatically active heterologous Rubisco protein in vivo by recombinant DNA procedures using large subunits of Anacystis nidulans and small subunits of Cylindrotheca sp. N1. It appears that two copies of rbcL and rbcS genes are encoded by the chloroplast DNA of this diatom. PMID- 2562762 TI - Cloning and sequence analysis of cDNAs encoding the cytosolic precursors of subunits GapA and GapB of chloroplast glyceraldehyde-3-phosphate dehydrogenase from pea and spinach. AB - Chloroplast glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is composed of two different subunits, GapA and GapB. cDNA clones containing the entire coding sequences of the cytosolic precursors for GapA from pea and for GapB from pea and spinach have been identified, sequenced and the derived amino acid sequences have been compared to the corresponding sequences from tobacco, maize and mustard. These comparisons show that GapB differs from GapA in about 20% of its amino acid residues and by the presence of a flexible and negatively charged C-terminal extension, possibly responsible for the observed association of the enzyme with chloroplast envelopes in vitro. This C-terminal extension (29 or 30 residues) may be susceptible to proteolytic cleavage thereby leading to a conversion of chloroplast GAPDH isoenzyme I into isoenzyme II. Evolutionary rate comparisons at the amino acid sequence level show that chloroplast GapA and GapB evolve roughly two-fold slower than their cytosolic counterpart GapC. GapA and GapB transit peptides evolve about 10 times faster than the corresponding mature subunits. They are relatively long (68 and 83 residues for pea GapA and spinach GapB respectively) and share a similar amino acid framework with other chloroplast transit peptides. PMID- 2562763 TI - A cDNA-based comparison of dehydration-induced proteins (dehydrins) in barley and corn. AB - Several cDNAs related to an ABA-induced cDNA from barley aleurone were isolated from barley and corn seedlings that were undergoing dehydration. Four different barley polypeptides with sizes of 22.6, 16.2, 14.4 and 14.2 kDa and a single corn polypeptide with a size of 17.0 kDa were predicted from the nucleotide sequences of the cDNAs. These dehydration-induced proteins (dehydrins) are very similar to each other and to a previously identified rice protein induced by ABA and salt, and have at least some similarity to a previously identified cotton embryo protein. Each dehydrin is extremely hydrophilic, glycine-rich, cysteine- and tryptophan-free and contains repeated units in a conserved linear order. A lysine rich repeating unit occurs twice in each protein, once at the carboxy terminus and once partway through the polypeptide, adjacent to a succession of serines. This repeating unit and the adjacent flanking run of serines are conserved with minimal variation among all dehydrins. Another repeating unit is flanked by the two copies of the lysine-rich unit, and varies in number from one to five copies. This latter repeating unit is less conserved than the former, varying even within a singly dehydrin. The messenger RNAs corresponding to each cDNA are abundant in dehydrating, but not in well-watered seedlings. The amino acid sequence of tryptic peptides from purified dehydration-induced proteins of corn established that the corn cDNAs correspond to a protein that is produced in abundance during the response of corn seedlings to dehydration. PMID- 2562764 TI - Effects of in vitro lead exposure on voltage-sensitive calcium channels differ among cell types in central neurons of Lymnaea stagnalis. AB - The effects of acute in vitro lead exposure on slowly inactivating voltage sensitive calcium channels in central neurons of the freshwater pond snail Lymnaea stagnalis were studied under voltage clamp. Three physiologically distinct cell types were used: two subsets of the B cell cluster (Bpos and Bneg) and the pedal giant neuron (RPeD1). In Bpos neurons, 5 nM free Pb2+ irreversibly inhibited current flow through calcium channels by 38 +/- 10%. In Bneg neurons, 5 nM free Pb2+ slightly inhibited inward currents (12 +/- 6%) and may have shifted their voltage dependence to more depolarized voltages. The inhibition and voltage shift were irreversible. In RPeD1 neurons, Pb2+ caused a small, statistically insignificant inhibition of inward current (5 nM free Pb2+; 18 +/- 19%; 30 nM free Pb2+: 31 +/- 23%). The effects of Pb2+ were fully reversible. These data indicate that (1) voltage-sensitive calcium channels in Lymnaea neurons are inhibited by nanomolar concentrations of free Pb2+; (2) there are multiple types of calcium channels in Lymnaea neurons; and (3) the effects of in vitro lead exposure differ qualitatively among channel types. PMID- 2562765 TI - Effect of organic and inorganic mercuric salts on Na+K+ATPase in different cerebral fractions in control and intrauterine growth-retarded rats: alterations induced by serotonin. AB - An intrauterine growth-retarded (IUGR) model based on restriction of blood supply to the rat fetus at the 17th day of pregnancy was studied. We investigated in vitro the effects of thimerosal and mercuric chloride on Na+K+ATPase activity in total brain homogenate, synaptosomes and myelin at weaning. In addition, we evaluated the reversal effect of serotonin on mercury-inhibited Na+K+ATPase activity. The toxicity, in terms of inhibition of Na+K+ATPase activity was greater with mercuric chloride than with thimerosal. Synaptosomes and principally myelin were more sensitive to the metal salts than total homogenate. Serotonin stimulated the Na+K+ATPase activity in total brain homogenate and synaptosomes but inhibited the enzyme in the myelin fraction. This effect was more marked in the IUGR group than in the control group. Serotonin (1 mM) added to total homogenate pretreated with the mercury salts produced variable reversal effects. In the synaptosomal fraction reverse effect was noted with serotonin. In myelin fraction, added serotonin increased inhibition caused by thimerosal. PMID- 2562766 TI - Interaction of the pyrethroid insecticides tetramethrin and cypermethrin with enteric cholinergic transmission in the guinea-pig. AB - In electrically-stimulated longitudinal muscle-myenteric plexus preparations of the guinea-pig ileum, the Type I pyrethroid insecticide tetramethrin (1-100 microM) caused a biphasic response consisting of an early transient increase followed by a sustained decrease in the amplitude of cholinergic contractions. The cholinergic potentiation was antagonized by phenytoin (3 microM), which also prevented the increase in twitch height caused by veratridine (30 nM). The late inhibitory effect of tetramethrin probably involved a direct action on the musculature since contractile responses to applied acetylcholine (100 nM) or histamine (300 nM) were also depressed by this compound. Cypermethrin (1-100 microM), a Type II pyrethroid, had only a minor enhancing effect on electrically evoked contractions. Cypermethrin (30, 60 microM), but not tetramethrin, antagonized the cholinergic response induced by the GABA-A receptor agonist 3 aminopropane sulphonic acid (1-100 microM). These results suggest that neural Na+ channels activation may underlie pyrethroid-induced potentiation of enteric cholinergic transmission. In small intestine, however, cypermethrin is also effective as a noncompetitive antagonist of GABA-A receptor mediated cholinergic contractions. PMID- 2562767 TI - Myelinated nerve fiber regeneration following organophosphorus ester-induced delayed neuropathy. AB - Chickens which developed organophosphorus ester-induced delayed neuropathy (OPIDN) due to a single oral dose of 360 mg/kg tri-ortho-tolyl phosphate were followed for up to 64 days following toxicant administration. Neuropathy was well developed by day 14. Progressive, marked but incomplete clinical improvement was observed between that time and day 49, associated with regenerative process was initially noted on day 16 at a distal, non-terminal level of the tibial nerve branch to the lateral head of the gastrocnemius muscle. One or more axonal sprouts were seen in bands of Bungner, but only one regenerated fiber per band myelinated and grew. By day 64 the nerves from treated hens closely resembled those from the controls. Myelinated nerve fiber degeneration was noted in distal regions of long spinal cord white matter tracts, but no subsequent regeneration was observed in this region. These studies indicate that in OPIDN there is a transient period of neuronal injury, and that damaged cells having peripherally directed neurites are likely able to undertake axonal regeneration. PMID- 2562768 TI - Identification of a mitochondrial protein associated with cytoplasmic male sterility in petunia. AB - The petunia fused gene (pcf), which is associated with cytoplasmic male sterility (CMS), is composed of sequences derived from atp9, coxII, and an unidentified reading frame termed urfS. To determine whether the pcf gene is expressed at the protein level, we produced antibodies to synthetic peptides specified by the coxII and urfS portions of the pcf gene. Anti-COXII peptide antibodies recognized petunia COXII but no other mitochondrial proteins. Anti-URF-S peptide antibodies recognized a 20-kilodalton protein present in both cytoplasmic male sterile and fertile lines and a protein with an apparent molecular mass of 25 kilodaltons present only in cytoplasmic male sterile lines. The 25-kilodalton protein was found to be synthesized by isolated mitochondria and to fractionate into both the soluble and membrane portions of disrupted mitochondria, whereas the 20 kilodalton protein was found only in the membrane fraction. The abundance of the 25-kilodalton protein was much lower in fertile plants carrying the cytoplasmic male sterile cytoplasm and a single dominant nuclear fertility restorer gene, Rf. Thus, the pcf gene is correlated with cytoplasmic male sterility not only by its co-segregation with the phenotype in somatic hybrids, but also by the modification of its expression at the protein level through the action of a nuclear gene that confers fertility. PMID- 2562769 TI - [Study on the causes for some characteristic damages of diamond instruments for dental practice]. AB - An attempt has been made for investigation and classification of the causes for diamond instruments for dental practice resource decrease. The overheating of the instruments is considered to be the main cause and the plastic materials smudges and unstable fixing of the diamond grains--as accompanying causes. Taking in consideration the above mentioned factors could contribute to a more rational use of the diamond instruments for dental practice. PMID- 2562770 TI - [The role of neuro-psychic diseases in the etiology of periodontosis]. AB - Experimental, clinical and statistical studies on the effect and connection between the neuro-psychic diseases and the diseases of periodontium have been described and analyzed. Attention is drawn to the effect of the second signal system and stress states and diseases (with psychogenic, emotional character). The mechanism of stress effect is explained by the law of dialectic for unity and struggle of opposites. The possibility of stress states becoming etiological factors or morbid conditions for periodontium diseases is well grounded. PMID- 2562771 TI - [Some problems of clinical characteristics, diagnostics and treatment of pleomorphic adenoma of salivary glands]. AB - The author evaluates the clinical characteristics in the light of the concepts proceeding from the International histological classification of tumours of the salivary glands on the base of his own experience in the treatment of 208 patients with pleomorphic adenoma of the salivary glands. The existing hospital documentation of the treated patients was analyzed as well as a repeated evaluation of the histological diagnosis made during the treatment and control of the therapeutic results via periodical inquiry of the treated patients. The author confirms the ratified unified nomenclature of the International classification and the benign nature of the pleomorphic adenoma. In that aspect he defends the surgical treatment of the pleomorphic adenoma as the method of choice and the high diagnostic value of the urgent intraoperative histological study. He advances the thesis for the application of more radical methods in the surgical treatment of recurrent pleomorphic adenoma of the parotid gland. According to the author, the morphological characteristics of the pleomorphic adenoma have no effect on the choice of the surgical methods for the treatment of that tumour. PMID- 2562772 TI - [The effect of exposure to silicon dioxide on the occurrence of porphyria tarda]. AB - The incidence of porphyria cutanea tarda was studied in two groups of silicon dioxide risk workers and compared to that in a control group. In the group with higher exposure to SiO2 the illness occurred in 12 out of 440 workers, in the group with lower exposure in 12 out of 1000 workers. In the control group porphyria was found in 18 out of 12,100 individuals examined. The difference in the incidence of the illness between the SiO2 risk individuals and the control group is statistically significant (p less than 0.01), while the difference between the group with variously high SiO2 exposure is on the level of statistically significance (p less than 0.05). The causes of the higher incidence rate of porphyria cutanea tarda in free SiO2 risk workers are discussed. PMID- 2562773 TI - [Levels of ACTH and endogenous-digitalis-like substances in human blood]. AB - Simultaneous determination of immunoreactive ACTH and immunoreactive digitalis like substances (DLS) in 71 plasma specimens of 44 persons proved that in spite of a great range of ACTH concentrations in the followed individuals (1.7-271 nmol/l) there is not a major correlation between DLS and ACTH or a significant difference in plasma DLS concentrations in the groups of persons with suppressed, normal or increased ACTH concentrations. Acute increase of ACTH plasma concentrations after synthetic ACTH application or a suppression of plasma ACTH by dexamethasone were not accompanied by corresponding changes of immunoreactive DLS. Thus it follows from this investigation that in spite of its biological digitalis like activity and interference of synthetic ACTH 1-24 high concentrations with digoxin enzymoimmunoassay in vitro, ACTH in concentrations found in human plasma is not responsible for endogenous immunoreactive DLS levels in plasma. PMID- 2562774 TI - An alternative regimen of hormone replacement therapy to improve patient compliance. AB - In order to achieve the long-term benefits from hormone replacement therapy of a markedly reduced incidence of heart disease and osteoporosis, a high degree of compliance is essential. In an effort to obtain high compliance, it was decided to introduce a cyclic therapy of 6 months duration using conjugated oestrogens supported by a 10-day course of progestogens. A total of 85 patients were treated prospectively. Compliance was assessed by the number of patients continuing treatment, and endometrial response was assessed by office biopsy and cytological or histological examination. Five patients withdrew, giving an overall compliance rate of 94%. Two have subsequently resumed therapy, thus 98% of those enrolled are currently receiving hormone replacement therapy. No cases of endometrial carcinoma were detected during the trial period of 4 years. Two cases of mild atypical endometrial hyperplasia were detected but both were mild and reverted to secretory or inactive endometrium following progestogen therapy. This regimen provides a viable alternative for those women who are troubled by progestogenic side-effects and monthly withdrawal bleeding. PMID- 2562775 TI - Effects of PEG-coupled interleukin-2 on rat lung lavage parameters. AB - The recombinant lymphokine interleukin-2 (IL-2) has activity in renal cell carcinoma, melanoma, and other cancers. A side effect of IL-2 use is a "capillary leak phenomenon" which is purported to be related to endothelial effects of IL-2 itself or to cells activated by IL-2. We studied IL-2 effects on rat lung lavage parameters to determine whether endothelial damage occurred. The specific endpoints were 125I-albumin extravasation, lavage protein, and lavage angiotensin converting enzyme (ACE) activity. To ensure sensitivity of these endpoints, we used the known endothelial toxicant thiourea, which increases lung lavage protein and lavage ACE. We found that both PEG IL-2 and thiourea increased the amount of protein and 125-I flux into the lavage. However, although thiourea increased lavage ACE, PEG IL-2 did not. These results suggest that PEG IL-2 can increase protein and iodine flux across the endothelium without causing cell injury. PMID- 2562776 TI - Mammalian neuronal differentiation: early expression of a neuronal phenotype from mouse neural crest cells in a chemically defined culture medium. AB - We show that mouse neural crest cells cultured in a serum-deprived chemically defined medium on appropriate culture substrata can be induced to express a neuronal phenotype. The uncommitted neural crest cells express a mesenchymal intermediate filament protein such as vimentin, but not the usual neuronal markers such as receptor sites for tetanus toxin or neurofilaments. In the chemically defined medium, receptor sites for tetanus toxin or neurofilaments can be characterized after a few hours in culture. Furthermore, these cells acquire tetrodotoxin-sensitive voltage-dependent Na+ channels and can generate action potentials. Such an in vitro system should allow us to analyze and manipulate early stages of neuronal differentiation in a mammalian embryo, at a level so far restricted to lower vertebrate embryos. PMID- 2562777 TI - [Miliary lung metastases of hepatocarcinoma: a rare form of presentation of the disease]. AB - The radiological demonstration of lung metastasis in hepatocarcinoma is a rare manifestation. The occurrence in a miliary pattern fashion is exceptional and even more so if it is the initial clinical feature. A case with all these clinical features is presented. PMID- 2562778 TI - The maize transposable element Ac excises in progeny of transformed tobacco. AB - To assess the potential of the maize transposable element Ac for gene tagging in heterologous plant species we monitored transcription, excision and transposition of the element in transgenic tobacco plants and their selfed progeny. Ac excised in the majority of primary regenerants and continued to excise in the first generation progeny plants. In one primary regenerant Ac was transcribed but did not excise. Fourteen of eighteen kanamycin-resistant progeny from this plant showed Ac excision, suggesting that excision of Ac may have been activated during meiosis or in embryo development. This finding, together with the more general observation of continued Ac mobility in the progeny of transformed plants in which Ac had excised, suggests that Ac will be useful for gene tagging. PMID- 2562779 TI - Isolation and nucleotide sequence of the pea cytochrome oxidase subunit I gene. PMID- 2562782 TI - 3,7-dinitrofluoranthene. PMID- 2562783 TI - Management of birth asphyxia by traditional birth attendants. AB - Birth asphyxia is an important cause of perinatal mortality, especially in developing countries. A study in India has shown that traditional birth attendants can recognize the condition but mostly cannot deal with it. The authors suggest that this deficiency could be overcome if suitable training were given. PMID- 2562785 TI - Intrathoracic instillation of autologous blood in treating massive hydrothorax following CAPD. PMID- 2562784 TI - Microtubule-associated protein 1B: molecular structure, localization, and phosphorylation-dependent expression in developing neurons. AB - Two monoclonal antibodies, 5E6 and 1B6, were raised against microtubule associated protein 1B (MAP1B), a major component of the neuronal cytoskeleton. 5E6 recognized the entire MAP1B population, while 1B6 detected only phosphorylated forms. Affinity-purified MAP1B appeared as a long, filamentous molecule (186 +/- 38 nm) with a small spherical portion at one end, forming long cross-bridges between microtubules in vitro. These results, together with in vivo data from immunogold methods, demonstrate that MAP1B is a component of cross bridges between microtubules in neurons. By immunohistochemical analysis, phosphorylated forms were shown to exist mainly in axons, whereas unphosphorylated forms were limited to cell bodies and dendrites. Phosphorylated MAP1B was quite abundant in developing axons, suggesting its essential role in axonal elongation. PMID- 2562786 TI - Gold-naproxen pneumonitis. A toxic drug interaction? AB - A patient with rheumatoid arthritis developed restrictive lung disease and blood eosinophilia. Gold pneumonitis was suspected but the patient did not improve until naproxen was discontinued as well. Lymphocyte transformation studies suggested hypersensitivity to gold. We hypothesize that naproxen unmasked and perpetuated the manifestations of gold hypersensitivity in our patient. PMID- 2562788 TI - Prognostic significance of eosinophils and mast cells in rectal cancer: findings from the National Surgical Adjuvant Breast and Bowel Project (protocol R-01). AB - The numbers of eosinophils and mast cells observed at the tumor border of 331 rectal cancers from patients enrolled in the National Surgical Adjuvant Breast and Bowel Project (NSABP), protocol R-01, were correlated according to overall survival rate, as well as Dukes' stage, tumor differentiation, nodal status, degree of lymphoid and stromal reactions, sex, and age. Life table plots disclosed a significantly better overall survival rate when ten or more eosinophils per 30 oil immersion fields were found. However, the numbers of eosinophils were strongly associated with Dukes' stage and, when life table plots were adjusted for Dukes' stage, this relationship to survival rate was not evident. On the other hand, overall survival rate was significantly higher in patients in whom 0 to three mast cells per 30 oil immersion fields were found than in those patients in whom four or more mast cells were found. This relationship persisted even when life table plots were adjusted for treatment, Dukes' stage, or nodal status, and indicated that the number of mast cells further defined survival rate among patients exhibiting Dukes' A, B, and C stages. It is concluded that numbers of eosinophils and mast cells may play a role in the natural history of rectal cancer but only the latter represents a prognostic parameter independent of Dukes' stage or nodal status. The mechanism whereby mast cells may exert this effect is at present unknown. PMID- 2562789 TI - [Bacteriological, pharmacokinetic and clinical studies on cefpodoxime proxetil in the pediatric field]. AB - Bacteriological, pharmacokinetic and clinical studies on cefpodoxime proxetil (CPDX-PR, CS-807), a new oral cephem antibiotic, were carried out in the field of pediatrics. The results obtained are summarized as follows. 1. Antibacterial activities of R-3746 (Na-salt of cefpodoxime (CPDX] against clinically isolated strains of Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus, Enterococcus faecalis, Branhamella catarrhalis, Escherichia coli, Proteus mirabilis and Haemophilus influenzae were compared with those of cefaclor, cephalexin and cefadroxil. R-3746 is superior to other antibiotics against S. pneumoniae, S. pyogenes, B. catarrhalis and Gram-negative rods. 2. Serum concentrations of CPDX after administration of CPDX-PR at doses of 3 mg/kg (fasting), 6 mg/kg (non-fasting) and 6 mg/kg (fasting) were determined. Mean AUC (area under curve)'s of CPDX obtained were 9.60, 31.35 and 17.89 micrograms.hr/ml, respectively for the 3 dosages. The mean half-lives of CPDX were 3.35, 1.88 and 1.76 hours, respectively. The mean urinary recovery rate within 8 hours after administration of CPDX-PR at a dose of 3 mg/kg (fasting) was 39.2%. 3. CPDX-PR was administered to 37 pediatric patients with various bacterial infections (pyelonephritis 9, cystitis 4, pneumonia 7, acute bronchitis 3, otitis media 2, tonsillitis 10, subcutaneous abscess 1 and purulent lymphadenitis 1). The overall clinical efficacy rate was 91.9% and the overall bacteriological eradication rate was also 91.9%. 4. No adverse reactions were observed. Abnormal laboratory findings were moderate, eosinophilia in 2 and slight elevation of GOT and GPT in 1. The taste and the odor of the CPDX-PR preparation was sufficiently tolerable. From the above results we have concluded that CPDX-PR is a useful oral antibiotic in the treatment of bacterial infections in children. PMID- 2562787 TI - Identification of a zinc finger protein that binds to the sterol regulatory element. AB - Cholesterol balance in mammalian cells is maintained in part by sterol-mediated repression of gene transcription for the low density lipoprotein receptor and enzymes in the cholesterol biosynthetic pathway. A promoter sequence termed the sterol regulatory element (SRE) is essential for this repression. With the use of an oligonucleotide containing the SRE to screen a human hepatoma complementary DNA expression library, a clone for a DNA binding protein was isolated that binds to the conserved SRE octanucleotide in both a sequence-specific and a single strand--specific manner. This protein contains seven highly conserved zinc finger repeats that exhibit striking sequence similarity to retroviral nucleic acid binding proteins (NBPs). We have designated the protein "cellular NBP" (CNBP). CNBP is expressed in a wide variety of tissues, is up regulated by sterols, and exhibits binding specificity that correlates with in vivo function. These properties are consistent with a role in sterol-mediated control of transcription. PMID- 2562790 TI - [Clinical studies on cefpodoxime proxetil dry syrup in the field of pediatrics]. AB - Cefpodoxime proxetil (CPDX-PR, CS-807) dry syrup was administered orally to 31 patients with various infections at daily dose levels between 5.4 and 10.9 mg/kg divided into three doses. 1. The subjects were 3 patients with urinary tract infections, 25 with tonsillitis and 1 patient each with bronchitis, pneumonia, and cervical lymphadenitis. Clinical effects were excellent in 16 cases, good in 14, and fair in 1 (tonsillitis), with an overall efficacy rate of 96.8%. 2. Organisms suspected as pathogens were 32 strains (6 strains of Staphylococcus aureus, 2 of Streptococcus pyogenes, 1 of Enterococcus faecalis, 15 of Haemophilus influenzae, 5 of Haemophilus parainfluenzae and 3 of Escherichia coli). Bacteriologically, eradication of pathogens were observed in 30 strains, decrease in one (H. parainfluenzae), and no change in another (E. faecalis), thus the eradication rate was 93.8%. 3. Side effect was observed in 1 case (slight eruption) but it was possible continue the treatment. Abnormal laboratory test values were observed in 1 case of a slight prolongation of prothrombin time and eosinophilia, but they were not serious. Diarrhea was not observed in any patients. 4. All the medication was done on schedule. No refusal of the drug occurred due to its taste or odor. PMID- 2562791 TI - Prevention of aflatoxicosis by addition of hydrated sodium calcium aluminosilicate to the diets of growing barrows. AB - Hydrated sodium calcium aluminosilicate (HSCAS), an anticaking agent for mixed feed, was added to the diets of growing barrows and was evaluated for its potential to ameliorate the clinical signs of aflatoxicosis. The experimental design consisted of 6 treatments of 5 barrows each at concentrations of 0 g of HSCAS and 0 g of aflatoxin (AF)/kg of feed (control), 5 g of HSCAS/kg of feed (0.5%), 20 g of HSCAS/kg of feed (2.0%), 3 mg of AF/kg of feed, 5 g of HSCAS (0.5%) plus 3 mg of AF/kg of feed, or 20 g of HSCAS (2.0%) plus 3 mg of AF/kg of feed. Barrows were maintained in indoor concrete-floored pens, with feed and water available ad libitum for 28 days (from the age of 7 to 11 weeks). Barrows were observed twice daily and were weighed weekly, and blood samples were obtained weekly for hematologic and serum biochemical measurements. At the termination of the study, barrows were euthanatized and necropsied. Body weight gains were diminished significantly (P less than 0.05) by consumption of 3 mg of AF/kg of feed, whereas body weight gain in barrows consuming diets containing HSCAS or HSCAS plus AF did not differ from that in control barrows. Serum enzymatic activities of alkaline phosphatase and gamma-glutamyl transferase and prothrombin time were increased in barrows consuming 3 mg of AF/kg of feed, but not in those consuming HSCAS or HSCAS plus AF.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562792 TI - Risks from circumcision during the first month of life compared with those for uncircumcised boys. AB - The records of 136,086 boys born in US Army hospitals from 1980 to 1985 were reviewed for indexed complications related to circumcision status during the first month of life. For 100,157 circumcised boys, there were 193 complications (0.19%). These included 62 local infections, eight cases of bacteremia, 83 incidences of hemorrhage (31 requiring ligature and three requiring transfusion), 25 instances of surgical trauma, and 20 urinary tract infections. There were no deaths or reported losses of the glans or entire penis. By contrast, the complications in the 35,929 uncircumcised infants were all related to urinary tract infections. Of the 88 boys with such infections (0.24%), 32 had concomitant bacteremia, three had meningitis, two had renal failure, and two died. The frequencies of urinary tract infection (P less than .0001) and bacteremia (P less than .0002) were significantly higher in the uncircumcised boys. Serious complications from routine prepuce removal are rare and relatively minor. Circumcision may be beneficial in reducing the occurrence of urinary tract infections and their associated sequelae. PMID- 2562794 TI - Blood donor programs. PMID- 2562793 TI - More on right to life. PMID- 2562795 TI - Estrogen receptor expression in human breast cancer associated with an estrogen receptor gene restriction fragment length polymorphism. AB - Estrogen receptor (ER) content is a well-known predictor of clinical outcome in human breast cancer. The recent cloning of a human ER complementary DNA has made possible the characterization of the ER gene on a molecular level. We have examined in human breast cancers a single, two-allele restriction fragment length polymorphism using the restriction enzyme PvuII. Initial studies in human breast cancer cell lines suggested a possible association between the absence of one allele and the absence of ER expression; subsequent analysis of allele distribution and frequency in 188 primary human breast tumor biopsies did indeed show a significant but not complete correlation between the absence of one allele and the failure to express ER. Preliminary data suggest that this restriction fragment length polymorphism is located within gene sequences coding for the putative DNA or hormone-binding domains of the ER. PMID- 2562796 TI - Defective in vitro migratory capacity of bone marrow cells from viable motheaten mice in response to normal thymus culture supernatants. AB - "Viable motheaten" mice are severely immunodeficient and develop autoantibodies early in life. The thymus appears normal for the first 3-4 weeks, after which there is depletion of cortical thymocytes and a diminution in the size of the organ until it is atrophic. The present study utilized an in vitro migration assay, in which bone marrow cells from viable motheaten mice were found to have a greatly diminished capacity to migrate in response to normal thymus supernatant when compared to normal bone marrow cells. It was also determined that thymus supernatant prepared from newborn viable motheaten mice was chemoattractive to normal bone marrow but not to viable motheaten bone marrow. The results of in vivo reconstitution of lethally irradiated viable motheaten mice with normal bone marrow cells also show that the thymus of the mutant is normal in its ability to attract and be repopulated by normal donor bone marrow. Therefore, the premature thymic involution of viable motheaten mice is related to the inability of bone marrow cells from these mice to migrate or respond to signals from the thymus. PMID- 2562798 TI - Hepatitis B-associated polyarteritis nodosa in Alaskan Eskimos: clinical and epidemiologic features and long-term follow-up. AB - We analyzed the demographic, clinical, laboratory and histologic features of 13 patients who were diagnosed as having polyarteritis nodosa associated with hepatitis B virus infection over a 12-year period, 1974 to 1985. All 13 patients were Yupik Eskimos and resided in southwest Alaska, an area hyperendemic for hepatitis B virus infection. The annual incidence of hepatitis B virus-associated polyarteritis nodosa for this population is 7.7 cases per 100,000 population. All patients presented with multisystem disease, and all had biopsy or angiographic findings consistent with polyarteritis nodosa. All 13 were positive for hepatitis B surface antigen and hepatitis B e antigen at diagnosis. Two untreated patients and two of five patients who received corticosteroids died, vs. none of six who received corticosteroids plus cyclophosphamide. None of the patients who survived the initial bout of polyarteritis nodosa has relapsed after a mean follow-up of 55 months, but all have become chronic HBsAg carriers. In eight patients, clinical or serologic evidence indicated that polyarteritis nodosa followed recent hepatitis B virus infection. We concluded that hepatitis B virus associated polyarteritis nodosa is a serious, life-threatening complication that occurs early in the course of hepatitis B virus infection, is ameliorated by immunosuppressive therapy and can be prevented by hepatitis B vaccine. PMID- 2562797 TI - Different capacities for amino acid transport in periportal and perivenous hepatocytes isolated by digitonin/collagenase perfusion. AB - Periportal and perivenous hepatocytes were isolated from rat liver by digitonin/collagenase perfusion for investigating the acinar heterogeneity of amino acid transport activities related to glutamine and ammonia metabolism. Immunocytochemical staining of the respective subpopulations for glutamine synthetase demonstrated that periportal subpopulations were essentially free of glutamine synthetase-positive cells, whereas perivenous subpopulations showed a 2 to 3-fold enrichment of glutamine synthetase-positive hepatocytes. The high perivenous/periportal ratio of 59 found for glutamine synthetase activity as well as the perivenous/periportal ratios of other marker enzymes further indicated the good separation of periportal and perivenous cells. alpha-Aminoisobutyric acid, histidine and glutamate were used to determine the distribution pattern of amino acid transport systems A, N and G-, as well as of the sodium-independent uptake of these compounds 1 hr after isolation and after maximal hormonal stimulation during primary culture. The strong heterogeneity of the sodium-independent transport of histidine, characterized by higher perivenous transport rates [perivenous/periportal ratio: 1.5 (1 hr) to 3.5 (48 hr)], suggests a significant role of facilitated diffusion, presumably in glutamine export. Conversely, the strong heterogeneity of the sodium-dependent glutamate transport (System G-) characterized by higher uptake rates in nonstimulated [perivenous/periportal ratio: 6.6 (1 hr)] and in hormonally treated perivenous hepatocytes (perivenous/periportal ratio: 2.2) reflects its possible significance with respect to the substrate availability for glutamine synthesis. The observed heterogeneities provide a basis for understanding how substrate fluxes related to glutamine metabolism might be established and regulated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562800 TI - A testis-specific gene Tpx-1 maps between Pgk-2 and Mep-1 on mouse chromosome 17. PMID- 2562799 TI - Limited MHC polymorphism in whales. AB - Little is known about disease and genetic variation in aquatic mammalian species such as whales. In this paper human HLA class I and class II probes were used to study major histocompatibility complex (MHC) genes from two species of whale: Fin (Balaenoptera physalus) and Sei (B. borealis). Stronger signals were obtained on whale than on equivalent concentrations of mouse DNA. Evidence was obtained for several DRB-related genes, a DNA genes, one DQA gene, and multiple class I genes in whales. Interestingly, the whale genes, from the small panel studied, were less polymorphic than those of humans or mice. The aquatic environment of this mammalian species may be a unique factor in shaping its immune response through the MHC. PMID- 2562801 TI - The germline repertoire of T cell receptor beta-chain genes in patients with chronic progressive multiple sclerosis. AB - The T cell receptor (TcR) beta-chain germline gene repertoire of multiple sclerosis (MS) patients was compared to that of 100 normal individuals. No differences in the number of gene segments defined by probes representing 14 different human V beta subfamilies and the constant region genes were found. The distribution of haplotypes defined by restriction fragment length polymorphism (RFLP) alleles detected with V beta 8, V beta 11, and C beta probes in the MS patients was significantly different from that found in normal individuals. Because 84% of the MS patients were DR2+, the findings in these patients were compared to a second group of 43 normals who were DR2+. The distribution of TcR haplotypes in MS patients was also significantly different from that in the DR2+ normals. The data suggest that an MS susceptibility gene(s) may be located in the region of the TcR beta-chain gene complex. PMID- 2562802 TI - Regulation of tyrosine hydroxylase mRNA in catecholaminergic cells of embryonic rat: analysis by in situ hybridization. AB - In situ hybridization was used to examine the appearance of mRNA specific for tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine (CA) biosynthesis, in neural crest derivatives of the rat embryo. These derivatives include sympathetic ganglia and transient catecholaminergic cells of embryonic intestine. Messenger RNA is first detected in sympathetic ganglia at E11.5, the age corresponding to the initial immunocytochemical expression of TH protein. In older embryos increased accumulation of TH-specific mRNA in sympathetic ganglia parallels the increase in TH immunoreactivity. By contrast, mRNA for TH is difficult to detect in embryonic intestines at E11.5 but is found instead in cells clustered at the dorsal boundaries of the pharynx and foregut. Cells expressing TH mRNA are infrequently found in embryonic intestines at any age, even though TH protein is immunohistochemically apparent. Treatment of pregnant rats with doses of reserpine, known to increase circulating levels of glucocorticoid hormones and prolong the expression of TH protein in embryonic gut cells, dramatically but transiently increases the number of gut cells at E12.5 with detectable TH mRNA. After E13.5 TH mRNA is undetectable even in reserpine treated guts. Reserpine treatment also increases the labeling density in sympathetic ganglia. Taken together, these data are consistent with the hypothesis that the microenvironment of the embryonic intestine affects gene expression directly to alter phenotype. Moreover, although reserpine administration briefly increases TH mRNA levels, the effect is short-lived and does not alter neurotransmitter phenotypic conversion. PMID- 2562804 TI - Stimulation of the Na+/K+ pump activity during electrogenic uptake of acidic amino acid transmitters by rat brain synaptosomes. AB - Addition of D-aspartate, a substrate for the high-affinity transport of acidic amino acid transmitters, to suspensions of rat brain synaptosomes increased the rate of O2 consumption, uptake of 86Rb, and transport of 2-[3H]deoxyglucose. Stimulation of all three processes was abolished in the presence of ouabain. D Aspartate had no effect on respiration in the medium in which NaCl was replaced by choline chloride. The ratio of the ouabain-sensitive increase in 86Rb uptake to that in O2 consumption was 12 to 1, which gives a calculated 86Rb(K+)/ATP of 2. It is concluded that electrogenic, high-affinity transport of sodium-D aspartate into synaptosomes stimulates the activity of the Na+/K+ pump through an increase in [Na+]i. PMID- 2562803 TI - Expression of excitatory amino acid receptors by cerebellar cells of the type-2 astrocyte cell lineage. AB - We have used postnatal rat cerebellar astrocyte-enriched cultures to study the excitatory amino acid receptors present on these cells. In the cultures used, type-2 astrocytes (recognized by the monoclonal antibodies A2B5 and LB1) selectively took up gamma-[3H]aminobutyric acid ([3H]GABA) and released it when incubated in the presence of micromolar concentrations of kainic and quisqualic acids. The releasing effect of kainic acid was concentration dependent in the range of 5-100 microM. Quisqualate was more effective than kainate in the lower concentration range but less effective at concentrations at which its releasing activity was maximal (approximately 50 microM). N-Methyl-D-aspartic acid and dihydrokainate (100 microM) did not stimulate [3H]GABA release from cultured astrocytes. L-Glutamic acid (20-100 microM) stimulated [3H]GABA release as effectively as kainate. The stimulatory effects of kainate and quisqualate on [3H]GABA release were completely Na+ dependent; that of kainate was also partially Ca2+ dependent. Kynurenic acid (50-200 microM) selectively antagonized the releasing effects of kainic acid and also that of L-glutamate; quisqualate was unaffected. Quisqualic acid inhibited the releasing effects of kainic acid when both agonists were used at equimolar concentrations (50 microM). D [3H]aspartate was taken up by both type-1 and type-2 astrocytes, but only type-2 astrocytes released it in the presence of kainic acid. Excitatory amino acid receptors with a pharmacology similar to that of the receptors present in type-2 astrocytes were also expressed by the immature, bipotential progenitors of type-2 astrocytes and oligodendrocytes. PMID- 2562805 TI - Dendrotoxin, 4-aminopyridine, and beta-bungarotoxin act at common loci but by two distinct mechanisms to induce Ca2+-dependent release of glutamate from guinea-pig cerebrocortical synaptosomes. AB - The release of endogenous glutamate from guinea-pig cerebrocortical synaptosomes evoked by dendrotoxin, beta-bungarotoxin, and 4-aminopyridine is compared. Dendrotoxin and 4-aminopyridine cause Ca2+-dependent release, representing a partial depletion of the KCl-releasable transmitter pool. The decrease in the plasma membrane potential caused by 4-aminopyridine or dendrotoxin and the evoked release of glutamate from a transmitter pool accord with the inhibitory action of these agents on certain K+ conductances. In contrast, the massive release of glutamate evoked by beta-bungarotoxin is produced in the presence of Ca2+ but not of Sr2+, a result consistent with a generalised permeabilisation of synaptosomal plasma membranes. Although dendrotoxin inhibits the binding of beta-bungarotoxin and the resultant synaptosomal lysis, demonstration of a direct effect of beta bungarotoxin binding per se on K+ permeability is impractical owing to its phospholipase A2 activity. PMID- 2562806 TI - Phosphorylation of B-50 (GAP43) is correlated with neurotransmitter release in rat hippocampal slices. AB - Recent studies have demonstrated that phorbol diesters enhance the release of various neurotransmitters. It is generally accepted that activation of protein kinase C (PKC) is the mechanism by which phorbol diesters act on neurotransmitter release. The action of PKC in neurotransmitter release is very likely mediated by phosphorylation of substrate proteins localized in the presynaptic nerve terminal. An important presynaptic substrate of PKC is B-50. To investigate whether B-50 mediates the actions of PKC in neurotransmitter release, we have studied B-50 phosphorylation in intact rat hippocampal slices under conditions that stimulate or inhibit PKC and neurotransmitter release. The slices were labelled with [32P]orthophosphate. After treatment, the slices were homogenized, B-50 was immunoprecipitated from the slice homogenate, and the incorporation of 32P into B-50 was determined. Chemical depolarization (30 mM K+) and the presence of phorbol diesters, conditions that stimulate neurotransmitter release, separately and in combination, also enhance B-50 phosphorylation. Polymyxin B, an inhibitor of PKC and neurotransmitter release, decreases concentration dependently the depolarization-induced stimulation of B-50 phosphorylation. The effects of depolarization are not detectable at low extracellular Ca2+ concentrations. It is concluded that in rat hippocampal slices B-50 may mediate the action of PKC in neurotransmitter release. PMID- 2562807 TI - Insulin binding to human astrocytoma cells and its effect on uridine incorporation into nucleic acid. AB - Binding of [125I]monoiodoinsulin to human astrocytoma cells (U-373 MG) was time dependent, reaching equilibrium after 1 h at 22 degrees C with equilibrium binding corresponding to 2.2 fmol/mg protein: this represents approximately 2,000 occupied binding sites per cell. The t1/2 of 125I-insulin dissociation at 22 degrees C was 10 min; the dissociation rate constant of 1.1 X 10(-2) s-1 was unaffected by a high concentration of unlabeled insulin (16.7 microM). Porcine insulin competed for specific 125I-insulin binding in a dose-dependent manner and Scatchard analysis suggested multiple affinity binding sites (higher affinity Ka = 4.4 X 10(8) M-1 and lower affinity Ka = 7.4 X 10(6) M-1). Glucagon and somatostatin did not compete for specific insulin binding. Incubation of cells with insulin (0.5 microM) for 2 h at 37 degrees C increased [2-14C]uridine incorporation into nucleic acid by 62 +/- 2% (n = 3) above basal. Cyclic AMP, in the absence of insulin, also stimulated nucleoside incorporation into nucleic acid [65 +/- 1% (n = 3)] above basal. Preincubation with cyclic AMP followed by insulin had an additive effect on nucleoside incorporation [160 +/- 4% (n = 3) above basal]. Dipyridamole (50 microM), a nucleoside transport inhibitor, blocked both basal and stimulated uridine incorporation. These studies confirm that human astrocytoma cells possess specific insulin receptors with a demonstrable effect of ligand binding on uridine incorporation into nucleic acid. PMID- 2562808 TI - Release by electrical stimulation of endogenous glutamate, gamma-aminobutyric acid, and other amino acids from slices of the rat medulla oblongata. AB - Evidence was obtained for the release of amino acids by electrical stimulation of slices of regions of the rat medulla oblongata: rostral ventrolateral, caudal ventrolateral and caudal dorsomedial. There was a Ca2+-dependent, tetrodotoxin sensitive increase in the efflux of aspartate, glutamate, gamma-aminobutyric acid (GABA), glycine, and beta-alanine in all regions examined. There were distinct regional differences in the relative amounts of amino acids released. These results provide evidence for the possible neurotransmitter role of aspartate, glutamate, GABA, glycine, and beta-alanine in these regions of the rat medulla oblongata. PMID- 2562809 TI - In vitro phosphorylation of bovine adrenal chromaffin cell tyrosine hydroxylase by endogenous protein kinases. AB - Under phosphorylating conditions, addition of Ca2+ or cyclic AMP to the 100,000 g supernatant of purified bovine adrenal chromaffin cells increases both the incorporation of 32P into tyrosine hydroxylase and the activity of the enzyme. Combining maximally effective concentrations of each of these stimulating agents produces an additive increase in both the level of 32P incorporation into tyrosine hydroxylase and the degree of activation of the enzyme. The increased phosphorylation by Ca2+ is due to stimulation of endogenous Ca2+-dependent protein kinase activity and not inhibition of phosphoprotein phosphatases. When the chromaffin cell supernatant is subjected to diethylaminoethyl (DEAE) chromatography to remove calmodulin and phospholipids, tyrosine hydroxylase is no longer phosphorylated or activated by Ca2+; on the other hand, phosphorylation and activation of tyrosine hydroxylase by cyclic AMP are not affected. Subsequent replacement of either Ca2+ plus calmodulin or Ca2+ plus phosphatidylserine to the DEAE-fractionated cell supernatant restores the phosphorylation, but not activation of the enzyme. Reverse-phase HPLC peptide mapping of tryptic digests of tyrosine hydroxylase from the 100,000 g supernatant shows that the Ca2+ dependent phosphorylation occurs on three phosphopeptides, whereas the cyclic AMP dependent phosphorylation occurs on one of these peptides. In the DEAE preparation, either cyclic AMP alone or Ca2+ in the presence of phosphatidylserine stimulates the phosphorylation of only a single phosphopeptide peak, the same peptide phosphorylated by cyclic AMP in the crude supernatant. In contrast, Ca2+ in the presence of calmodulin stimulates the phosphorylation of three peptides having reverse-phase HPLC retention times that are identical to peptides phosphorylated by Ca2+ addition to the crude unfractionated 100,000 g supernatant. Rechromatography of the peaks from each of the in vitro phosphorylations, either in combination with each other or in combination with each of the seven peaks generated from phosphorylation of tyrosine hydroxylase in situ, established that cyclic AMP, Ca2+/phosphatidylserine, and Ca2+/calmodulin all stimulate the phosphorylation of the same reverse-phase HPLC peptide: in situ peptide 6. Ca2+/calmodulin stimulates the phosphorylation of in situ peptides 3 and 5 as well. Thus, tyrosine hydroxylase can be phosphorylated in vitro by protein kinases endogenous to the chromaffin cell. Phosphorylation occurs on a maximum of three of the seven in situ phosphorylated sites, and all three of these sites can be phosphorylated by a Ca2+/calmodulin-dependent protein kinase. PMID- 2562810 TI - Phosphorylation of tubulin by casein kinase II regulates its binding to a neuronal protein (NP 185) associated with brain coated vesicles. AB - We recently described a new protein associated exclusively with neuronal clathrin coated vesicles (CCVs), and characterized two monoclonal antibodies that react with it (S-8G8 and S-6G7). In this report, the association of neuronal protein of 185 kilodaltons (NP185) with CCV kinases and its interaction with tubulin are described. The affinity of NP185 for tubulin is significantly enhanced when tubulin is phosphorylated by CCV-associated casein kinase II. In contrast, phosphorylation of tubulin by a kinase activity associated with purified brain tubulin decreases its affinity for NP185. Together, these data suggest that the interaction of NP185 with tubulin is modulated by protein phosphorylation. Recent evidence has suggested that tubulin is phosphorylated by casein kinase II during neurite development. The enhanced affinity of NP185 for tubulin phosphorylated by casein kinase II could be important for proper intracellular sorting of this protein in the developing neuron. PMID- 2562811 TI - Effects of DL-2-amino-5-phosphonovalerate on metabolism of catecholamines in synaptosomes from rat brain. AB - Incubation of synaptosomes from rat brain with DL-2-amino-5-phosphonovalerate (APV) stimulated an increased release of dopamine, and this effect was strictly dependent on the extrasynaptosomal calcium level. APV increased biosynthesis of dopamine from tyrosine by 30%, whereas monoamine oxidase activity was inhibited by 30%. When synaptosomes were incubated with radioactive dopamine, APV caused a large decrease in incorporation of label into 3,4-dihydroxyphenylacetic acid but greatly increased incorporation into norepinephrine and its N-methyl derivatives. Quantification of dopamine and its metabolites in synaptosomes, using electrochemical detection, indicated that the presence of APV resulted in changes in the absolute levels of the aforementioned dopamine metabolites similar to the changes in radiolabel incorporation. Omission of Ca2+ from the extrasynaptosomal medium greatly diminished the APV-induced changes in catecholamine metabolism. The metabolic changes appear to largely result from an increased intrasynaptosomal Ca2+ level due to the APV-induced increase in calcium permeability of the plasma membrane. PMID- 2562812 TI - Characterization of cysteine proteases functioning in degradation of dynorphin in neuroblastoma cells: evidence for the presence of a novel enzyme with strict specificity toward paired basic residues. AB - Two dynorphin-degrading cysteine proteases, I and II, were extracted with Triton X-100 from neuroblastoma cell membrane, isolated from accompanying dynorphin degrading trypsin-like enzyme by affinity chromatography on columns of soybean trypsin inhibitor-immobilized Sepharose and p-mercuribenzoate-Sepharose, and separated by ion-exchange chromatography on diethylaminoethyl (DEAE)-cellulose and TSK gel DEAE-5PW columns. Cysteine protease II was purified further by hydroxyapatite chromatography and gel filtration. The molecular weights of cysteine proteases I and II were estimated to be 100,000 and 70,000, respectively, by gel filtration. Both of the enzymes, were inhibited by p chloromercuribenzoate, N-ethylmaleimide, and high-molecular-weight kininogen, but not or only slightly inhibited by diisopropylphosphorofluoridate, antipain, leupeptin, E-64, calpain inhibitor, and phosphoramidon. Cysteine protease I cleaved dynorphin(1-17) at the Arg6-Arg7 bond with the optimum pH of 8.0, whereas II cleaved dynorphin(1-17) at the Lys11-Leu12 bond and the Leu12-Lys13 bond with the optimum pH values of 8.0 and 6.0, respectively. These bonds corresponded to those that had been proposed as the initial sites of degradation by neuroblastoma cell membrane. Cysteine protease I was further found to show strict specificity toward the Arg-Arg doublet, when susceptibilities of various peptides containing paired basic residues were examined as substrates for the enzyme. PMID- 2562813 TI - Spontaneous rearrangement of integrated simian virus 40 DNA in nine transformed rodent cell lines. AB - Frequencies of spontaneous DNA rearrangement within or near integrated simian virus 40 (SV40) DNA were measured in four transformed mouse and rat cell lines of independent origin and in five clones of the SV40-transformed mouse line SVT2. Rearrangements were detected as polymorphisms of restriction enzyme fragment length in subclones of the lines. At least 17% of the subclones of each line had detectable rearrangements. The rate of rearrangement was calculated to be at least 5 x 10(-3) events per cell per division. No rearrangements were detected in sequences of an immunoglobulin gene, part of the coding region of the mouse protein p53, and five proto-oncogenes. The possible role of recombination between duplicated segments of integrated SV40 DNA in generating rearrangements was studied in the five SVT2 clones, which differed in the number of duplications within a single SV40 DNA segment. The SVT2 clone that had no duplications, M3, became rearranged further at least as frequently as did closely related lines with one, two, or three duplications. Another line in this group that had one small duplication, X1, had a much higher frequency of rearrangement than did the others; integrated SV40 DNA of X1 became mostly rearranged within 100 cell divisions. The examples of M3 and X1 suggested that the high rate of rearrangement characteristic of integrated SV40 DNA was influenced more by the presence of particular sequences within or near integrated SV40 DNA than by the number or extent of duplicated sequences. PMID- 2562815 TI - Effect of acute complete obstruction on the rabbit urinary bladder. AB - Studies on the effect of partial bladder outlet obstruction demonstrate that significant alterations in urinary bladder structure and function occur within 24 hours of the creation of the obstruction. These studies suggest that many of the functional and structural alterations following partial outlet obstruction may result from the initial overdistension which occurs within the first 24 hours. In these present studies, we investigated the time course of the effect of complete obstruction of the rabbit urinary bladder on the contractile response to bethanechol and field stimulation and on the muscarinic receptor density. Male White New Zealand rabbits were divided into five groups: controls, four, eight, 20, and 24 hours of complete obstruction. The results demonstrated that there was a significant decrease in both the contractile response to muscarinic stimulation and muscarinic receptor density at four hours following outlet obstruction (at a time when there was no bladder overdistension). The receptor density and contractile response to stimulation further decreased over the 24 hour period. These studies indicate that the initial decrease in muscarinic receptor density and contractile response to muscarinic stimulation may be mediated in part by the high level of spontaneous contractile activity induced by ligation of the urethra. PMID- 2562814 TI - Prion protein biosynthesis in scrapie-infected and uninfected neuroblastoma cells. AB - Numerous studies have indicated that a modified proteinase K-resistant form of an endogenous brain protein, prion protein (PrP), is associated with scrapie infection in animals. This scrapie-associated PrP modification appears to occur posttranslationally in brain, but its molecular nature is not known. To learn about the normal PrP biosynthesis and whether it is altered by scrapie infection in vitro, we did metabolic labeling experiments with uninfected and scrapie infected mouse neuroblastoma tissue culture cells. Pulse-chase labeling experiments indicated that, in both cell types, two major PrP precursors of 28 and 33 kilodaltons (kDa) were processed to mature 30- and 35- to 41-kDa forms. Endoglycosidase H, tunicamycin, and phospholipase treatments revealed that the 28 and 33-kDa precursors resulted from the addition of high-mannose glycans to a 25 kDa polypeptide containing a phosphatidylinositol moiety and that maturation of the precursors involved the conversion of the high-mannose glycans to hybrid or complex glycans. Treatments of the live cells with trypsin and phosphatidylinositol-specific phospholipase C indicated that the mature PrP species were expressed solely on the cell surface, where they were anchored by covalent linkage to phosphatidylinositol. Once on the cell surface, the major PrP forms had half-lives of 3 to 6 h. No differences in PrP biosynthesis were observed between the scrapie-infected versus uninfected neuroblastoma cells. PMID- 2562816 TI - Effects of age on urinary bladder function in the male rat. AB - In a previous study we investigated the effects of age on the micturition characteristics and bladder function of male Fischer rats ages five to seven, 16 to 18 and 22 to 24 months. The 24 hr. water intake and urine output increases significantly with age; 22 to 24 month rats showed a 39% increase in water intake and a 93% increase in urine output compared to five to seven month rats. The intravesical pressure at micturition is 100% greater in 22 to 24 month and 16 to 18 month rats compared to five to seven month old rats with no age-related change in bladder volume at micturition. In the present study, in vitro bladder capacity did not differ between the three age groups although the average plateau pressure significantly decreased with advancing age. Using the isolated whole bladder model, the contractile response to the autonomic agonists bethanechol, phenylephrine, and isoproterenol did not change significantly with age. Similarly, there were no age-related changes in the response of the bladder to non-autonomic drugs (histamine, oxytocin, serotonin, substance P, and PGF2 alpha) except for PGF2 alpha which produced an age-related increase in the maximum bladder contraction. In summary, while in vivo micturition clearly changes with age, the in vitro contractility of the bladders to autonomic agents did not. Therefore, age related differences in micturition would be related primarily to the changes in neuronal innervation and central control of micturition rather than alterations in the contractility of the bladder. In addition, these studies show the importance of correlating in vivo bladder function (micturition frequency and volume, cystometry and urodynamics) with in vitro contractile and functional studies. PMID- 2562817 TI - Myocardial properties of the new dihydropyridine calcium antagonist isradipine compared to nifedipine with or without additional beta blockade in coronary artery disease. AB - Isradipine is a new dihydropyridine calcium antagonist with myocardial effects significantly different from those of nifedipine, as shown by in vitro and animal experimental data. Isradipine selectively inhibits the sinus node but not the atrioventricular conduction and its negative inotropic action is much less if administered in a dose of comparable peripheral effects. To study these effects in man, 40 patients with coronary artery disease were divided into 2 groups receiving either a continuous 30-minute intravenous infusion of 2 mg of nifedipine or 0.5 mg of isradipine, doses that resulted in a comparable afterload reduction (decrease of systemic vascular resistance: nifedipine -22.1%, isradipine -25%, p less than 0.001). Ten patients in each group received an additional intravenous bolus of 5 mg of propranolol at the end of the calcium antagonist administration to antagonize its induced adrenergic reflex mechanisms. The heart rate significantly increased after nifedipine only (+9.2%, p less than 0.001), experienced no change after isradipine and the nifedipine and propranolol combination and decreased after the combination of isradipine and propranolol ( 9.6%, p less than 0.001). This resulted in a significant decrease of the rate pressure product with isradipine (-12.5%, p less than 0.001) but not with nifedipine. As a result of the afterload-induced adrenergic reflex mechanisms, the maximal derivative of the left ventricular pressure increased after isradipine administration (+13.5%, p less than 0.001) and was unchanged after nifedipine, which demonstrates the significantly less negative inotropic properties of isradipine as compared with nifedipine. PMID- 2562818 TI - Prevalence of cardiac abnormalities in human immunodeficiency virus infection. AB - The prevalence of cardiac abnormalities in the spectrum of human immunodeficiency virus (HIV) infection is unknown. Sixty consecutive HIV-infected patients were studied using echocardiograms, electrocardiograms (50 patients) and ambulatory electrocardiographic monitoring (43 patients). Group A (25 patients) were seropositive but pre-AIDS, whereas group B (35 patients) had AIDS and included 24 with an active opportunistic infection (group B1) and 11 without it (group B2). Abnormalities were identified in 32 of 60 patients (53%) and were more frequent in group B (23 of 35, 66%) than in group A (9 of 25, 36%, p less than 0.05) but independent of active opportunistic infection (15 of 24, 62%, in group B1 vs 8 of 11, 73%, in group B2). Echocardiographic abnormalities were identified in 21 of 60 patients (35%), including 7 of 25 (28%) in group A vs 14 of 35 (40%) in group B (difference not significant), and 7 of 24 (29%) in group B1 vs 7 of 11 (64%) in group B2 (difference not significant). Those patients with an absolute CD4 lymphocyte count less than or equal to 100/mm3 had a higher prevalence of echocardiographic abnormalities (12 of 22) than those with CD4 counts greater than 100/mm3 (1 of 14, p less than 0.01). Left ventricular dilation or hypokinesis was identified in 14 of 60 patients (23%), including 4 of 25 (16%) in group A and 10 of 35 (29%) in group B. Electrocardiographic abnormalities were seen in 22 of 50 patients (44%) including 5 of 18 (28%) in group A and 17 of 32 (53%) in group B (difference not significant).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562819 TI - Selection against lethal alleles in females heterozygous for incontinentia pigmenti. AB - Studies of five heterozygous females from three kindreds segregating incontinentia pigmenti indicate that cells expressing the mutation have been eliminated from skin fibroblast cultures and in varying degrees from hematopoietic tissues. Clonal analysis was carried out using G6PD variants and methylation patterns at the HPRT locus. Our results confirm X linkage in these families and suggest that selection against cells expressing mutations that are lethal to males in utero may help ameliorate the deleterious phenotype in carrier females. PMID- 2562820 TI - Linkage analysis of the apolipoprotein C2 gene and myotonic dystrophy on human chromosome 19 reveals linkage disequilibrium in a French-Canadian population. AB - The gene for human apolipoprotein C2 (APOC2), situated on the proximal long arm of chromosome 19, is closely linked to the gene for the most common form of adult muscular dystrophy, myotonic dystrophy (DM). Six APOC2 RFLPs (TaqI, BglI, BanI, BamHI, NcoI, and AvaII) have been identified to date. We have conducted a comprehensive DM linkage study utilizing all six RFLPs and involving 50 families and 372 individuals. The most informative RFLPs are, in descending order, NcoI (lod = 6.64, theta = 0.05), BglI (lod = 6.12, theta = 0.05), AvaII (lod = 6.02, theta = 0.03), BanI (lod = 5.76, theta = 0.04), TaqI (lod = 4.29, theta = 0.06), and BamHI (lod = 1.75, theta = 0.01). A substantial increase in the lod scores over those seen with the individual RFLPs was obtained when the linkage of the entire APOC2 haplotype (composed of the six RFLPs) was studied (lod = 17.87, theta = 0.04). We have observed significant inter-APOC2 RFLP linkage disequilibrium. Consequently, the three most informative RFLPs have been found to be BanI, TaqI, and either BglI, AvaII, or NcoI polymorphisms. We also demonstrate linkage disequilibrium between DM and APOC2 in our French-Canadian population (standardized disequilibrium constant phi = .22, chi 2 = 5.12, df = 1, P less than 0.04). This represents the first evidence of linkage disequilibrium between APOC2 and the DM locus. PMID- 2562821 TI - Lack of linkage disequilibrium between two common restriction sites associated with the COLIA2 gene. PMID- 2562822 TI - Fine structure DNA mapping studies of the chromosomal region harboring the genetic defect in neurofibromatosis type I. AB - To better map the location of the von Recklinghausen neurofibromatosis (NF1) gene, we have characterized a somatic cell hybrid designated 7AE-11. This microcell-mediated, chromosome-transfer construct harbors a centromeric segment and a neo-marked segment from the distal long arm of human chromosome 17. We have identified 269 cosmid clones with human sequences from a 7AE-11 library and, using a panel of somatic cell hybrids with a total of six chromosome 17q breakpoints, have mapped 240 of these clones on chromosome 17q. The panel included a hybrid (NF13) carrying a der(22) chromosome that was isolated from an NF1 patient with a balanced translocation, t(17;22) (q11.2;q11.2). Fifty-three of the cosmids map into a region spanning the NF13 breakpoint, as defined by the two closest flanking breakpoints (17q11.2 and 17q11.2-q12). RFLP clones from a subset of these cosmids have been mapped by linkage analysis in normal reference families, to localize the NF1 gene more precisely and to enhance the potential for genetic diagnosis of this disorder. The cosmids in the NF1 region will be an important resource for testing DNA blots of large-fragment restriction-enzyme digests from NF1 patient cell lines, to detect rearrangements in patients' DNA and to identify the 17;22 NF1 translocation breakpoint. PMID- 2562824 TI - Random tyrosine and glutamic acid-containing polymers are very powerful inhibitors of casein kinase-2. AB - The random heteropolymers Glu/Tyr(4:1) and Glu/Tyr(1:1) that are widely used as substrates for tyrosine protein kinases, are very powerful competitive inhibitors of casein kinase-2, but not of casein kinase-1, with respect to the protein substrate, their Ki values being one to two orders of magnitude lower than those of polyglutamates of similar size. The inhibitory power is reduced if tyrosine is partially replaced by alanine, as in the polymer Glu/Ala/Tyr(6:3:1) and it disappears upon inclusion of lysine, the polymer Glu/Ala/Lys/Tyr(2:6:5:1) actually behaving as a stimulator. These data indicate that non-phosphorylatable hydroxylic residues in addition to acidic ones are required in order to optimize the binding of pseudo-substrates to the catalytic site of casein kinase-2. PMID- 2562823 TI - Origin and differentiation of human mitochondrial DNA. AB - A recent study of mitochondrial DNA (mtDNA) polymorphism has generated much debate about modern human origins by proposing the existence of an "African Eve" living 200,000 years ago somewhere in Africa. In an attempt to synthesize information concerning human mtDNA genetic polymorphism, all available data on mtDNA RFLP have been gathered. A phylogeny of the mtDNA types found in 10 populations reveals that all types could have issued from a single common ancestral type. The distribution of shared types between continental groups indicates that caucasoid populations could be the closest to an ancestral population from which all other continental groups would have diverged. A partial phylogeny of the types found in five other populations also demonstrates that the myth of an African Eden was based on an incorrect "genealogical tree" of mtDNA types. Two measures of molecular diversity have been computed on all samples on the basis of mtDNA type frequencies, on one hand, and on the basis of the number of polymorphic sites in the samples, on the other. A large discrepancy is found between the two measures except in African populations; this suggests the existence of some differential selective mechanisms. The lapse of time necessary for creating the observed molecular diversity from an ancestral monomorphic population has been calculated and is found generally greater in Oriental and caucasoid populations. Implications concerning human mtDNA evolution are discussed. PMID- 2562825 TI - Comparison of respiratory effects of two cardioselective beta-blockers, celiprolol and atenolol, in asthmatics with mild to moderate hypertension. AB - The effects of two cardioselective beta-adrenergic blocking agents--celiprolol (claimed to have bronchodilator properties) and atenolol (without such claims)- on respiratory function and control of asthma were studied in ten asthmatic patients with mild to moderate essential hypertension. Following a beta-2 agonist free period of ten hours, administration of 100 mg of atenolol was associated with bronchoconstriction (p less than 0.05), whereas 400 mg of celiprolol was not. Responsiveness to beta-2 agonist therapy was retained with both agents (p less than 0.05). Day-to-day asthma control, interpreted from patient recordings of peak flow, inhaler use and symptom scores, were all no different on either agent from placebo. When given acutely, celiprolol appeared to have acute bronchosparing properties, possibly providing a greater margin of respiratory safety than atenolol. PMID- 2562826 TI - Growth hormone-releasing factor-sensitive adenylate cyclase system of purified somatotrophs: effects of guanine nucleotides, somatostatin, calcium, and magnesium. AB - The purpose of this study was to characterize the adenylate cyclase system in a purified population of normal somatotrophs derived from rat pituitary and to determine the responses of this system to GRF, somatostatin, guanine nucleotides, and cations. Additionally, experiments were performed to evaluate the interrelationships among changes in adenylate cyclase activity, cellular cAMP levels, and GH release induced by GRF and somatostatin. The results obtained using homogenates and membrane preparations from somatotrophs included the following. 1) GRF caused guanine nucleotide-dependent concentration-related (Ka, approximately 10(-8) M) stimulation of adenylate cyclase activity. 2) Guanine nucleotides were effective in stimulating cyclase in the absence of GRF; the concentration of guanine nucleotide required for half-maximal stimulation was decreased more than 10-fold in the presence of GRF. 3) Adenylate cyclase activity increased with increasing concentrations of free Mg2+ (0.25-20 mM); activation by GRF and guanine nucleotide resulted in an approximately 7-fold increase in the enzyme's affinity for free Mg2+. 4) Somatostatin, up to 10(-6) M, did not alter basal or GRF-stimulated adenylate cyclase activity. 5) Ca2+ (0.5-11.9 microM) produced concentration-dependent inhibition of basal (up to 28%) and GRF stimulated (up to 47%) cyclase activities; the inhibitory effect of Ca2+ was accompanied by a decrement (2- to 3-fold) in the apparent affinities of the enzyme for both GRF and guanine nucleotide. In intact somatotrophs, GRF produced concentration-dependent stimulation of GH release (Ka, approximately 6 x 10(-11) M), preceded by a marked elevation of cAMP levels. While somatostatin blocked GRF induced GH release, the augmented cAMP levels were only slightly reduced.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2562827 TI - Interleukin-1 beta stimulates somatostatin biosynthesis in primary cultures of fetal rat brain. AB - Interleukin-1 (IL-1), a secretory product of activated macrophages and many other cell types, is an important mediator of the acute phase reaction to infection and to endotoxin administration. Previous reports that GH and TSH secretion are decreased following injection of endotoxin or IL-1 led us to test the hypothesis that IL-1 acts by releasing increased amounts of somatostatin (SS), a hypothalamic factor inhibitory of both GH and TSH release. Primary cultures of dispersed fetal rat diencephalic cells were found to contain increasing amounts of immunoreactive SS in both cells and media after addition of recombinant human IL-1 beta. This increase was detectable at 24 hours and continued for up to 6 days, the longest time interval tested. Increased content of SS peptide was accompanied by marked increases in SS mRNA. These changes were dose-related, the lowest effective dose being 10(-10) M. In contrast to the long term response, exposure of the cells to IL-1 beta for one hour had only minimal stimulating effects on somatostatin release. These data indicate that IL-1 beta is neurotrophic for the somatostatinergic neuron, an action that may be responsible at least in part, for the neuroendocrine response to infection. PMID- 2562828 TI - Regulation of growth hormone (GH) secretion by GH-releasing factor, somatostatin, and insulin-like growth factor I in ovine fetal and neonatal pituitary cells in vitro. AB - Serum GH concentrations in the ovine fetus are much higher than those in the neonate, and the maximal GH response induced by GRF is 5-fold greater in the fetus than in the neonate. To clarify these in vivo observations further, we studied the effects of GRF, somatostatin (SRIF), and insulin-like growth factor I (IGF-I) on primary cultures of fetal and neonatal ovine pituitary cells. GH secretion from fetal ovine pituitary cells increased from 148 +/- 34 to 950 +/- 130 ng/10(5) cells.3 h in response to 1 nM GRF, whereas GH secretion from neonatal pituitary cells rose from 113 +/- 26 to 1221 +/- 129 ng/10(5) cells.3 h, a significantly greater response (P less than 0.001). This greater GRF-induced GH response in neonatal than fetal cells differs from the response in vivo and suggests that the increased in vivo response in the fetus is not due to inherently increased sensitivity of pituitary cells to GRF. SRIF (10 nM) decreased maximal GRF-induced GH secretion by 37 +/- 3% in fetal cells compared with 59 +/- 8% in neonatal cells (P less than 0.01). This may explain in part the decreased in vivo sensitivity to SRIF in the ovine fetus compared to that in the neonatal lamb. In fetal pituitary cells, 10 nM GRF increased ovine (o) GH mRNA from 100 +/- 14% to 145 +/- 40%, SRIF decreased oGH mRNA to 84 +/- 3%, and GRF and SRIF in combination increased fetal oGH mRNA to 126 +/- 24%. Values in neonatal pituitary cell cultures were similar (control, 100 +/- 17%; GRF, 132 +/- 6%, SRIF, 85 +/- 15%; GRF plus SRIF, 105 +/- 26%). Pretreating fetal cells with 100 nM IGF-I for 3 days reduced GRF-stimulated GH secretion from 1049 +/- 38 to 232 +/- 8 ng/10(5) cells.3 h (P less than 0.001). Similarly, IGF-I pretreatment of neonatal cells reduced GRF-stimulated GH secretion from 810 +/- 18 to 419 +/- 16 ng/10(5) cells.3 h (P less than 0.001). The mean secreted IGF-I was 0.58 U/ml (36 nM) in culture medium from neonatal cells and was unchanged by incubation for 3 days with 5 micrograms/ml hGH. Secreted IGF-I in medium from fetal cells was 0.87 U/ml (54 nM) without GH and 0.81 U/ml (51 nM) after incubation with human GH. IGF-I mRNA was present in neonatal pituitary and brain.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2562829 TI - Positive and negative regulation of a transfected chimeric tyrosine aminotransferase gene: effect of copy number. AB - To define a selective system for the study of rat tyrosine aminotransferase (TAT; EC 2.6.1.5) gene expression, we have introduced into cultured cells the selectable bacterial gene gpt linked to TAT gene flanking sequences. After integration in host cell DNA, the chimeric gene exhibits the same pattern of regulation as the TAT gene. In hepatoma cells, its expression is induced after glucocorticoid hormone treatment and repressed after fusion with fibroblasts. In fibroblasts, the chimeric gene is not expressed. The correct pattern of regulation is lost when the number of integrated copies is high. At copy number above 10, the transfected gene responds poorly to glucocorticoids in hepatoma cells. At copy number above 50, the gene is expressed in fibroblasts. Another gene present in the same construction and controlled by the SV40 early promoter and enhancer is positively regulated by glucocorticoids in hepatoma cells but not after fusion with fibroblasts. These data indicate that in hybrid cells, both TAT promoter and glucocorticoid-responsive elements are negatively regulated. PMID- 2562830 TI - Sea urchin primary mesenchyme cells: ingression occurs independent of microtubules. AB - The formation of primary mesenchyme cells in euechinoid sea urchin embryos involves the transformation of 32 epithelial cells into mesenchymal cells in a process referred to as ingression. The mechanism that drives this epithelial mesenchymal transformation has yet to be identified. Previous studies (J. R. Gibbins, L. G. Tilney, and K. R. Porter, 1969, J. Cell Biol. 41, 201-226; L. G. Tilney and J. R. Gibbins, 1969, J. Cell Biol. 41, 227-250) implicated that microtubules are essential components for the normal development, including ingression, of the mesenchymal cells. In the present study I have reinvestigated the role of microtubules in ingression by using the microtubule-disrupting drugs colchicine and nocodazole, and the microtubule-stabilizing drug taxol. The effect of these drugs on microtubules was monitored by indirect immunofluorescence using monoclonal antibodies specific for alpha- and beta-tubulins. The microtubule array seen in control embryos disappeared in colchicine- and nocodazole-treated embryos, while it was enhanced in taxol-treated embryos. When premesenchyme blastulae of Strongylocentrotus purpuratus were treated with any of these reagents the primary mesenchyme cells ingressed on schedule and appeared to undergo cell-shape changes identical to those observed in untreated embryos. The conclusion of this study is that the mechanism of primary mesenchyme cell ingression does not include an essential role for microtubules; ingression occurs regardless of the presence or absence of microtubules. PMID- 2562831 TI - Insulin-receptor and apolipoprotein genes contribute to development of NIDDM in Chinese Americans. AB - The frequencies of restriction-fragment-length polymorphism (RFLP) alleles as well as RFLP haplotypes at six genetic loci responsible for carbohydrate and lipid metabolism [insulin/insulin-like growth factor II complex, insulin receptor (INSR), HepG2/erythrocyte-type glucose transporter, apolipoprotein A-II, apolipoprotein B (APOB), and the apolipoprotein A-I/C-III/A-IV cluster (APOA1/C3/A4)] were compared between nondiabetic and diabetic Chinese Americans. The disease-association data suggest that genetic variation at the INSR, APOB, and APOA1/C3/A4 loci contributes to the development of non-insulin-dependent diabetes mellitus (NIDDM). The analysis of the INSR locus revealed "protective" haplotypes, and it may be possible to use two of the INSR haplotypes as genetic markers to identify individuals having a very low probability of developing NIDDM regardless of the presence of other genes conferring susceptibility to this disorder. The APOB and APOA1/C3/A4 loci appear to contribute to the development of NIDDM in individuals who are of lean/normal weight and overweight, respectively. The APOA1/C3/A4 locus may account for approximately 8% of the difference between baseline and total possible risk of NIDDM in overweight individuals. PMID- 2562832 TI - Insulin-receptor gene and its expression in patients with insulin resistance. AB - We studied the structure of the insulin-receptor gene in normal individuals and in four unrelated patients with leprechaunism (Minn-1, Ark-1, Ark-2, Can-1) and four unrelated patients with the type A syndrome of insulin resistance, both disorders associated with genetic alterations in affinity, binding capacity, and kinase activity of the insulin receptor. Genomic cloning and Southern blot analysis indicate that the normal human insulin-receptor gene is greater than or equal to 150 kilobases long and consists of a minimum of 17 exons, 6 in the genomic region of the alpha-subunit and 11 in the region of the beta-subunit. Three of the patients, one with leprechaunism and two with type A syndrome, have decreases in insulin-receptor mRNA but on genomic blot analysis have no obvious abnormalities in the insulin-receptor gene. No distinctive pattern of restriction fragment-length polymorphisms or evidence for major insertion or deletion mutations of the insulin-receptor gene was found in any of the patients. These data indicate that the insulin-receptor gene is greater than 35 times larger than coding regions and has a complex structure. Although leprechaunism and type A syndrome are most likely due to defects in the structure and expression of the insulin-receptor gene, they are likely to be associated with specific point mutations rather than major changes in gene structure. PMID- 2562833 TI - Neural reflex of the canine pylorus to intraduodenal acid infusion. AB - In 29 chloralose-urethane anesthetized dogs, a manometric assembly was inserted through a gastrostomy to monitor pressure of the pyloric region with a sleeve sensor. Antral and duodenal contractions were monitored with both manometric sideholes and serosal strain gauges. An additional tube channel allowed intraduodenal infusions 1-2 cm aborad from the pylorus. Intraluminal infusion of hydrochloric acid (0.1 N, 0.92 ml/min, for 2 min) reproducibly caused activation of motor activity in the pyloric region and peristaltic duodenal activity. Proximal duodenal activity probably contributed to the total phasic response recorded in the pylorus region. Excitatory responses could also be elicited by infusion of phenyl-biguanide (stimulant of sensory nerve endings), but not by control infusions with diluent (Krebs' buffer or saline). The motor response of the pyloric region to intraduodenal acid was blocked by intraduodenal application of 2% xylocaine. Atropine (30 micrograms/kg i.v. and 100 micrograms i.a.) or hexamethonium (10 mg/kg and 1 mg i.a.) markedly reduced or blocked the acid induced pyloric motor response of this region but propranolol (1.0 mg/kg i.v. and 100 micrograms i.a.), phentolamine (1.5 mg/kg i.v. and 100 micrograms i.a.), or naloxone (200 micrograms/kg and 20 micrograms i.a.) had no effect. We believe these observations show the existence of a reflex from the duodenum to the pylorus in response to intraluminal stimuli mediated by a chain of cholinergic nerves. In the dog, endogenous opioid peptides do not contribute to the excitatory reflex pathway activated by intraduodenal acid or phenyl-biguanide. As intraluminal acid in the duodenum activates this reflex, it may play a role in the physiologic and pathophysiologic role of gastric emptying in this species. PMID- 2562834 TI - Effects of somatostatin on hepatic bile formation. AB - Somatostatin is a peptide that has anticholeretic properties in the dog. The purpose of the present work was to investigate if somatostatin is an anticholeretic agent in humans also. The effects of intravenous infusion of somatostatin on hepatic bile flow and biliary electrolytes and secretion of biliary lipids were studied in 7 patients with complete biliary drainage who had been operated on for choledocholithiasis. Somatostatin, 250 microgram/h, was found to decrease the hepatic bile secretion by approximately 30%. The peptide also reduced the outputs of bile acids, cholesterol, and phospholipids and the outputs of sodium, potassium, and chloride. The concentrations of the biliary lipids were not significantly changed. Somatostatin inhibited the erythritol clearance in the 2 patients studied by approximately 25%. The present study thus provides evidence that somatostatin inhibits bile formation in humans. It appears as if the reduction in bile production is mainly due to decreased canalicular bile flow. It is possible that this effect of somatostatin is attributable to inhibition of bile acid synthesis or of transport-secretion of bile acids, or both. PMID- 2562835 TI - Type 3 fimbriae among enterobacteria and the ability of spermidine to inhibit MR/K hemagglutination. AB - The distribution of the gene cluster encoding type 3 fimbriae among various isolates of the family Enterobacteriaceae was investigated by using 112 clinical and nonclinical isolates. Closely related DNA sequences were detected in all Klebsiella strains, in most Enterobacter isolates, in a smaller number of Escherichia coli and Salmonella spp., and in a single isolate each of Yersinia enterocolitica and Serratia liquefaciens but not in isolates of Morganella or Providencia species or Serratia marcescens. Except for E. coli and Salmonella strains, the presence of gene sequences was correlated with the phenotypic expression of either the MR/K hemagglutinin or fimbriae that reacted with specific antibodies. In one isolate of Y. enterocolitica the expression of type 3 fimbriae was plasmid determined. The polyamine spermidine was identified as an inhibitor of MR/K hemagglutinating activity, exhibiting an MIC of 1.2 mM. Spermidine inhibited the hemagglutination of 37 MR/K-positive clinical isolates from various genera. However, one clinical isolate of Enterobacter cloacae and most (four of five) nonclinical Klebsiella isolates were not completely inhibited. PMID- 2562837 TI - Age-specific colonization of porcine intestinal epithelium by 987P-piliated enterotoxigenic Escherichia coli. AB - Neonatal (less than 1-day-old), 3- and 7-day old, and older (3-week-old postweaning) pigs were challenged by intragastric inoculation with 987P-piliated (987P+) enterotoxigenic Escherichia coli (ETEC) 987. Neonatal pigs were colonized (i.e., there were greater than or equal to 10(8) CFU of test strain per 10-cm ileal segment) and developed diarrhea. Intestinal colonization and the incidence and severity of diarrhea were lower in 3- and 7-day old pigs than in neonates. Older pigs were not colonized and did not develop diarrhea following oral inoculation with five strains of 987P+ ETEC. Strain 987 (987P+) adhered in vitro to intestinal epithelial cell brush borders isolated from both neonatal (sensitive) and older (resistant) pigs. The in vivo growth and expression of 987P pilus by strain 987 in ligated ileal loops created in neonatal and older pigs were similar. The in vivo adherence of 987P+ ETEC to intestinal epithelium in ligated ileal loops in neonatal and older pigs was compared. In neonatal pigs, most of the bacteria were in layers associated with the villous epithelium. In older pigs, most of the bacteria were associated with mucus-like material in the intestinal lumen. We concluded that swine develop an innate resistance to 987P+ ETEC by 3 weeks of age. This resistance does not appear to be due to an absence of 987P-specific receptors in the intestines of the older pig or to an inability of 987P+ bacteria to grow and express pili in the older pig. We hypothesized that the resistance of older pigs to 987P-mediated disease is due to release of 987P specific receptors into the intestinal lumen, where these receptors facilitate bacterial clearance rather than bacterial adherence to intestinal epithelium and colonization. PMID- 2562836 TI - Isolation and characterization of the alpha-galactosyl-1,4-beta-galactosyl specific adhesin (P adhesin) from fimbriated Escherichia coli. AB - The alpha-galactosyl-1,4-beta-galactosyl-specific adhesin (P adhesin) was isolated from the fimbria-adhesin complex (FAC) of recombinant Escherichia coli strains expressing the F7(1), F8, or F13 fimbrial antigens. Separation into fimbriae and adhesin was achieved by heating the FAC to 80 degrees C in the presence of Zwittergent 3-16. After removal of the fimbriae by precipitation with lithium chloride, the adhesin was purified by anion-exchange fast protein liquid chromatography in the presence of 4 M urea. The purified adhesins from the three strains had pIs of 4.8 to 5.0 and molecular weights of approximately 35,000. The fimbrillins were smaller, their molecular weights being different with different F antigens. The amino-terminal amino acid sequence of the F7(1)- and F13-derived adhesins were different, that of the F13-derived adhesin being identical to that extrapolated from the DNA sequence of the papG gene (B. Lund, G. Lindberg, B.-I. Marklund, and S. Normark, Proc. Natl. Acad. Sci. USA 84:5898-5902). An antiadhesive monoclonal antibody which reacted with the three P adhesins was prepared. The FAC and the purified adhesins but not the fimbriae from which the adhesins had been removed agglutinated erythrocytes and galactose-galactose coated latex beads. The adhesion of erythrocytes to the surface-fixed adhesins could be specifically inhibited with alpha-galactosyl-1,4-beta-galactosyl-1,4 glucosyl. The results indicate that the P adhesin(s) of uropathogenic E. coli represents a group of related proteins with conserved receptor recognition domains. The F13-derived P adhesin is the PapG protein postulated by Normark and his colleagues (Lund et al., Proc. Natl. Acad. Sci. USA 84:5898-5902; B. Lund, F. Lindberg, and S. Normark, J. Bacteriol. 170:1887-1894). PMID- 2562838 TI - Palytoxin down-modulates the epidermal growth factor receptor through a sodium dependent pathway. AB - Palytoxin, a non-12-O-tetradecanoylphorbol-13-acetate type tumor promoter, has been shown to inhibit epidermal growth factor (EGF) binding to both high and low affinity receptors through a protein kinase C-independent pathway. In the present paper, we have investigated the mechanism of palytoxin action in Swiss 3T3 cells. Two lines of evidence indicate that calcium is not required for palytoxin activity. First, palytoxin can induce the loss of EGF binding sites in the absence of external calcium. Second, studies with the photosensitive protein aequorin indicate that palytoxin does not cause the influx of external calcium or the release of calcium from internal stores under the conditions used in these studies. However, palytoxin action does appear to be dependent upon the presence of sodium. When extracellular sodium is replaced by either choline, Tris, or sucrose, palytoxin is unable to decrease EGF binding to either high or low affinity receptors. Studies of sodium influx indicate that palytoxin induces rapid sodium uptake and that the rate of sodium uptake is dose-dependent. Furthermore, there appears to be a direct correspondence between the extent of inhibition of EGF binding by palytoxin and the rate of sodium uptake. Finally, the palytoxin-induced inhibition of EGF binding can be mimicked by monensin, a sodium ionophore. The specificity of this sodium dependence was tested by substituting lithium, potassium, or cesium for sodium. Although lithium is an effective substitute for sodium, palytoxin can no longer inhibit EGF binding when sodium is replaced by either potassium or cesium. Marked inhibition of palytoxin action is also obtained when 5.4 mM potassium or 5.4 mM cesium are added to the sodium-containing medium. These studies suggest that palytoxin is able to down modulate the EGF receptor through a novel mechanism involving the activation or formation of a sodium pump or channel. PMID- 2562839 TI - Dimeric dermorphin analogues as mu-receptor probes on rat brain membranes. Correlation between central mu-receptor potency and suppression of gastric acid secretion. AB - The opioid receptor preference for dermorphin and several dimerized structural analogues was investigated using rat brain synaptosomes and correlated with the potencies of intracerebroventricularly administered dimeric dermorphin peptides to inhibit gastric acid secretion. The carboxyl terminus of dermorphin or amino terminal dermorphin analogues was bridged by dihydrazide or (poly)ethylenediamine structures. Synaptosomal membranes were prepared for radioligand binding assay in the presence of soybean trypsin inhibitor and preincubated to remove endogenously bound opioid peptides before storage at -70 degrees C. Specific radiolabeled agonists used in the radioligand binding assays were [D-Ala2,N-methyl-Phe4,Gly ol5] [3H] enkephalin for mu-receptors and [D-Ala2,D-Leu5] [3H]enkephalin for delta-receptors. delta-Receptor binding assays were conducted in the presence of 2.6 microM [N-Me-Phe3,D-Pro4]morphiceptin to suppress peptide binding to mu receptors. [D-Ala2,N-methyl-Phe4,Gly-ol5]enkephalin and dermorphin had affinities of 1.39 and 1.22 nM for mu-receptors and 355.8 and 178.6 nM for delta-receptors, respectively. Affinities of dimeric-dermorphin0 for mu- and delta-receptors, and the mu-selectivity ratio, exceeded values characteristic of dermorphin. The dimerized amino-terminal dermorphin analogues are peptides whose receptor binding differed from the parent molecule; e.g. the affinity of dimeric tetrapeptides toward mu-receptors was reduced but was increased for delta-receptors relative to monomeric dermorphin-(1-4)-amide. Dimeric tetradermorphin linked by a bridge containing 12 methylene units (di-tetra-dermorphin12), exhibited a dramatic loss in the mu-selectivity ratio as a result of diminished mu-affinity. On the other hand, substitution of Gly4 by Sar in di-tetra-dermorphin2 enhanced binding to mu receptors: substitution of D-Arg2 for D-Ala resulted in an increased binding to mu-receptors while decreasing binding to delta-receptors, yielding a peptide with the highest mu-selectivity ratio. These substitutions of D-Arg2 and Sar4 in dimeric amino-terminal dermorphin pentapeptides enhanced binding to both mu- and delta-receptors relative to dermorphin-(1-5)-amide, but led to a decrease in its mu-selectivity ratio. Several dimeric dermorphin analogues exhibited an enhanced mu-selectivity ratio relative to their monomeric analogues. Dimeric peptides, which had a relatively high affinity for mu-receptors, were effective in the suppression of gastric acid secretion. PMID- 2562840 TI - The structure of tyrosine aminotransferase. Evidence for domains involved in catalysis and enzyme turnover. AB - The primary structure of tyrosine aminotransferase, as deduced from the nucleotide sequence of complementary DNA, was confirmed by fast atom bombardment mass spectrometry of tryptic peptides derived from the purified protein. Limited digestion of the native enzyme with trypsin released an acetylated, amino terminal peptide; the new amino terminus in the modified enzyme was Val65. Endogenous proteases generated a chromatographically separable form of tyrosine aminotransferase that began at Lys35. Neither trypsin nor the other proteases altered the catalytic activity of tyrosine aminotransferase. Reduction of the holoenzyme with sodium borohydride yielded a major tryptic peptide containing phosphopyridoxamine bound to lysine 280, which probably functions in transamination. The carboxyl terminus of tyrosine aminotransferase contains features that typify proteins with short half-lives; it includes two negatively charged, hydrophilic segments that are enriched for glutamyl residues and are similar to a PEST region in ornithine decarboxylase (Rogers, S., Wells, R., and Rechsteiner, M. (1986) Science 234, 364-368). Tyrosine aminotransferase belongs to a superfamily of enzymes which includes aspartate aminotransferase and can be aligned so that many invariant, functional residues coincide. Like the isoenzymes of aspartate aminotransferase, tyrosine aminotransferase may contain two domains, with a central, catalytic core, and a small domain made up of both amino- and carboxyl-terminal components. We speculate that the exposed small domain may confer the unusually rapid degradative rate that characterizes this enzyme. PMID- 2562841 TI - Inhibition of atrial natriuretic peptide-induced cyclic GMP accumulation in the bovine endothelial cells with anti-atrial natriuretic peptide receptor antiserum. AB - Using an antiserum raised against the purified atrial natriuretic peptide (ANP) receptor that has a disulfide-linked homodimeric structure and represents one subtype of the multiple ANP receptors, we showed that the receptor is coupled to the guanylate cyclase activation; formerly, this type of ANP receptor is not considered to be coupled to the cyclase. The specificity of the antiserum was determined by immunoblot analysis and immunoprecipitation. The anti-receptor antiserum did not compete with 125I-ANP for binding to the receptor but it lowered the affinity of the receptor. When added to bovine endothelial cell cultures, the antiserum blocked the cyclic GMP response of the cells triggered by ANP. These results indicate that the subtype of the ANP receptor recognized by the antiserum is responsible for the activation of particulate guanylate cyclase as well as the double function type receptor that has been assumed to contain both the receptor domain and the catalytic domain for cGMP synthesis on the same molecule. The presence of dissociative complexes of ANP receptor and particulate guanylate cyclase was also demonstrated by radiation inactivation analysis. PMID- 2562842 TI - Altered beta-adrenoceptor internalization does not explain reduced beta adrenergic responsiveness in the elderly. AB - To determine whether altered beta-adrenoceptor internalization contributes to the reduced adrenergic responsiveness of the elderly, the ability to internalize beta adrenoceptors in young and elderly men was studied by in vitro exposure of whole lymphocytes to a range of isoproterenol concentrations. The total numbers of receptors per cell were similar in the two groups [young, 1645 +/- 206 +/- SE); elderly, 1707 +/- 120], as were the basal levels of beta-adrenoceptor internalization (young, 8.85 +/- 1.09%; elderly, 8.73 +/- 1.35%). Incubation in the presence of isoproterenol resulted in dose-dependent internalization of receptors. The EC50 values for isoproterenol-induced internalization were similar in the young (20.7 +/- 3.5 nmol/L) and elderly groups (26.9 +/- 4.4 nmol/L). These results suggest that altered receptor internalization does not contribute to impaired beta-adrenoceptor function in the elderly. PMID- 2562843 TI - The persistent Mullerian duct syndrome: a molecular approach. AB - A rare form of male pseudohermaphroditism is characterized by the persistence of Mullerian derivatives in phenotypic males. To determine the etiology of this syndrome, we studied the expression of anti-Mullerian hormone (AMH) in six boys, including three brothers, with the persistent Mullerian duct syndrome. All except one presented with an inguinal hernia containing the Mullerian derivatives, and in two boys the hernial sac contained the contralateral testis. AMH was normally expressed in the testicular tissue of two patients, as shown by bioassay of anti Mullerian activity and immunocytochemistry. The testicular tissue of the other patients had no detectable bioactive or immunoreactive AMH, yet they expressed AMH mRNA with a normal transcription initiation site and in the amount expected for their age. These results prove the heterogeneity of the persistent Mullerian duct syndrome and suggest that it may sometimes involve peripheral insensitivity to AMH. PMID- 2562844 TI - Comparative hemodynamic and neurohormonal effects of intravenous captopril and digoxin and their combinations in patients with severe heart failure. AB - The effects of intravenous captopril and intravenous digoxin given separately and in combination on rest and exercise hemodynamics were studied in 16 patients with severe heart failure and sinus rhythm. When given separately, both captopril and digoxin decreased the pulmonary capillary wedge pressure by, respectively, 24% (p = 0.003) and 34% (p = 0.004) and systemic vascular resistance by 23% (p = 0.09) and 20% (p = 0.03). Only digoxin increased cardiac index by 23% (p = 0.03) and stroke work index by 52% (p = 0.01). During maximal exercise, captopril alone decreased systemic vascular resistance by 28% (p = 0.0002) and increased cardiac index by 33% (p = 0.02). Digoxin alone decreased pulmonary capillary wedge pressure by 11% (p = 0.04) and increased stroke work index by 44% (p = 0.01). The combination of captopril and digoxin resulted in a decrease in pulmonary capillary wedge pressure and systemic vascular resistance and an increase in cardiac index and stroke work index both at rest and during exercise that was greater than values observed with either drug given alone. Cardiac index response to the combination of captopril and digoxin correlated with baseline serum aldosterone concentration (r = 0.81, p less than 0.001) and plasma renin activity (r = 0.74, p less than 0.0002). A significant decrease in norepinephrine concentration was noted after digoxin was administered alone or added to captopril. These findings demonstrate that in patients with severe heart failure, the acute administration of captopril and digoxin has an independent salutary hemodynamic effect. The combination of these agents, however, has an adjunctive effect on cardiac function at rest and during exercise. PMID- 2562845 TI - ACC anniversary seminar. An editor's account of the history of the College. PMID- 2562846 TI - Restricted tissue distribution of Mlsa determinants. Stimulation of Mlsa-reactive T cells by B cells but not by dendritic cells or macrophages. AB - Evidence was sought on the tissue distribution of Mlsa determinants, a class of cell-associated non-H-2 alloantigens that is highly immunogenic for unprimed T cells. Whereas normal CD4+ T cells and an Mlsa-reactive T hybridoma gave strong responses to Mlsa-positive stimulator populations containing Ig+ B cells, anti Mlsa responses to B-depleted stimulators were almost undetectable. The B-depleted stimulators tested included Thy-1- spleen cells from mu-suppressed mice (mice treated with anti-mu antibody from birth) and J11d- preparations of spleen dendritic cells (DC) and peritoneal macrophages (M phi) from normal mice. Each of these populations was strongly immunogenic for allo-H-2-reactive T cells. The failure to detect Mlsa determinants on Ig- APC, i.e., M phi and DC, suggests that Mlsa determinants are not typical H-2-associated peptides. The data are more compatible with a model in which Mlsa determinants represent (or form part of) an integral cell membrane molecule expressed largely, and perhaps exclusively, on B cells. T cells might recognize these molecules only in native form, "processed" Mlsa determinants being nonimmunogenic. Consistent with this possibility, no evidence was found that Mlsa-negative B cells could absorb Mlsa determinants from Mlsa-positive B cells in a chimeric environment. PMID- 2562847 TI - Inhibition of cellular DNA synthesis by reovirus occurs through a receptor-linked signaling pathway that is mimicked by antiidiotypic, antireceptor antibody. AB - Mammalian reovirus type 3 binds to a 67-kD surface glycoprotein on the membrane of neuronal cells. This interaction initiates the infective reovirus cycle. The physiological function of this virus receptor is not known, however, initial studies illustrate a striking structural and antigenic homology to the beta adrenergic receptor family. The earliest known pathologic effect of reovirus type 3 is an inhibition of host cell DNA synthesis within 8-10 h after virus attachment. The studies reported here demonstrate that binding and aggregation of reovirus receptor molecules provides the signal for this inhibitory process. Inhibition of DNA synthesis in the neuroblastoma cell line B104.G4 was unaffected by using replication-defective virus or when lysosomal processing of normal virus was blocked. Inhibition was mimicked by an antiidiotypic, antireceptor mAb. Inhibition was not observed when monovalent mAb fragments were bound to receptors, but was reconstituted when these fragments were aggregated by the addition of anti-Ig. The signal for the inhibitory effect was generated within the first 60 min after mAb binding. These observations demonstrate that reovirus and antiidiotypic pathogenicity can result from the perturbation of membrane proteins that may perform normal physiological functions. PMID- 2562848 TI - Distinct features of dendritic cells and anti-Ig activated B cells as stimulators of the primary mixed leukocyte reaction. AB - Highly enriched populations of B lymphoblasts have been isolated after culture with anti-Ig-Sepharose and compared with dendritic cells as stimulators of CD4+ T cells in the murine MLR. The two populations clearly differed in phenotype; anti Ig blasts were FcR+, B220+, 33D1-, while dendritic cells were FcR-, B220-, 33D1+. However, as MLR stimulators, they shared many common features. Both cells (a) expressed comparable levels of class II MHC products; (b) independently stimulated the primary MLR and the production of several T derived lymphokines including IL-2 and IL-4; and (c) were comparable in stimulating freshly sensitized T cells. However, the relative potencies of dendritic cells and anti Ig blasts as primary MLR stimulators varied in a strain-dependent fashion. Only anti-Ig blasts could stimulate across an Mls barrier, being at least 100 times more active in stimulating Mls-mismatched, MHC-matched T cells, relative to syngeneic T cells. In contrast, dendritic cells were 10-30 times more potent than anti-Ig blasts when stimulating across an MHC barrier and were likewise more effective in binding MHC-disparate T cells to form the clusters in which the MLR was generated. Dendritic cell-T cell clustering was resistant to anti-LFA-1 mAb, while B blast-T cell clustering was totally blocked. Thus, anti-Ig B lymphoblasts and dendritic cells, two cell types which differ markedly in phenotype, also differ in efficiency and mechanism for initiating responses in allogeneic T cells. PMID- 2562850 TI - Insight in schizophrenia. Its relationship to acute psychopathology. AB - The relationship between insight and acute psychopathology was explored in a group of 52 acutely psychotic, schizophrenic patients. A measure of insight, reflecting patients' recognition of their illness and need for care, was validated against ratings from a semi-structured interview and against assessments of patients' compliance with medication. Contrary to expectations, degree of insight was not consistently related to the severity of acute psychopathology, as measured on two structured scales. Nor did changes in insight during hospitalization vary consistently with changes in acute psychopathology. These data suggest that very little of the deficiency in insight seen in schizophrenic patients is explainable on the basis of acute psychopathological features. The mechanism that accounts for impairment in insight in schizophrenia may be relatively resistant to treatment with neuroleptic medication. PMID- 2562849 TI - Clonal expansion of human T lymphocytes initiated by dendritic cells. AB - The accessory cell requirements for cloning T cells in the presence of lectin and T cell growth factors were examined with cells from human peripheral blood. We found that dendritic cells were active and perhaps essential. Single CD4+ lymphocytes could be cloned with 80% efficiency, and CD8+ cells with 50-60% efficiency if 10(3) syngeneic or allogeneic dendritic cells were added. Some T cell clones developed even with one dendritic cell. Monocytes or B lymphocytes from blood were at least 100-fold weaker in supporting clonal growth. These findings suggest a specialized feeder cell requirement, namely dendritic cells, for cloning T lymphocytes from single resting precursors. PMID- 2562851 TI - Teratogenic effects of benzodiazepine use during pregnancy. AB - Eight children exposed in utero to benzodiazepines had characteristic dysmorphic features, growth aberrations, and central nervous system abnormalities from birth. Their dysmorphic characteristics resembled those of the fetal alcohol syndrome, although they had greater focal involvement of cranial nerves, with a sullen and expressionless face, and they more often had impairment of vitality at birth. One infant died and at autopsy had varying degrees of distortion of neuronal migration, with concomitant heterotopias. Five of the eight mothers had regularly consumed benzodiazepines, and the three remaining mothers had blood samples during pregnancy revealing benzodiazepine concentrations indicative of regular use. Our findings indicate that maternal consumption of benzodiazepines may be teratogenic in humans. PMID- 2562852 TI - Synthesis and antiallergy activity of 4-(diarylhydroxymethyl)-1-[3 (aryloxy)propyl]piperidines and structurally related compounds. AB - A series of 4-(diarylhydroxymethyl)-1-[3-(aryloxy)propyl]piperidines was synthesized and evaluated for antiallergy activity. Several analogues had potent activity in the passive foot anaphylaxis (PFA) assay, an IgE-mediated model useful in the detection of compounds possessing antiallergic activity. In particular 1-[4-[3-[4-[bis(4-fluorophenyl)hydroxymethyl]-1-piperidinyl] propoxy] 3-methoxyphenyl]ethanone (1, AHR-5333) was more potent than oxatomide and terfenadine in this assay. PMID- 2562853 TI - Synthesis of an analogue of tabtoxinine as a potential inhibitor of D-alanine:D alanine ligase (ADP forming). AB - The design and synthesis of a potential inhibitor of D-alanine:D-alanine ligase (ADP forming) (EC 6.3.2.4) are described. This enzyme, which catalyzes the second step in the biosynthesis of bacterial peptidoglycan, is believed to generate D alanyl phosphate as an enzyme-bound intermediate. With tabtoxinine, a potent inhibitor of glutamine synthetase, as a model, beta-lactams 9R and 9S were synthesized as potential precursors of a D-alanyl phosphate mimic. PMID- 2562854 TI - Synthesis and alpha 2-adrenoceptor antagonist activity of some disulfonamidobenzoquinolizines. AB - A series of disulfonamidobenzo[a]quinolizines were synthesized and evaluated for their alpha 2- and alpha 1-adrenoceptor antagonist activity on the rat vas deferens and anococcygeus muscle, respectively. N-((2 beta,11b alpha) 1,3,4,6,7,11b-Hexahydro-2H-benzo[a]quinolizin-2-yl)-N- [2 [(methylsulfonyl)amino]ethyl]methanesulfonamide (4) and its N-[2 [(methylsulfonyl)amino]ethyl]ethanesulfonamide (22), N-[2 [(ethylsulfonyl)amino]ethyl]ethanesulfonamide (27), and N-[2 [(methylsulfonyl)amino]ethyl]-4-methylbenzenesulfonamide (30) analogues showed 400-fold or greater selectivity in favor of alpha 2- over alpha 1-adrenoceptor blockade. PMID- 2562856 TI - Expression of a multidrug resistance gene in human cancers. AB - Many cancers have been cured by chemotherapeutic agents. However, other cancers are intrinsically drug resistant, and some acquire resistance following chemotherapy. Cloning of the cDNA for the human MDR1 gene (also known as PGY1), which encodes the multidrug efflux protein P-glycoprotein, has made it possible to measure levels of MDR1 RNA in human cancers. We report the levels of MDR1 RNA in greater than 400 human cancers. MDR1 RNA levels were usually elevated in untreated, intrinsically drug-resistant tumors, including those derived from the colon, kidney, adrenal gland, liver, and pancreas, as well as in carcinoid tumors, chronic myelogenous leukemia in blast crisis, and cell lines of non-small cell carcinoma of the lung (NSCLC) with neuroendocrine properties. MDR1 RNA levels were occasionally elevated in other untreated cancers, including neuroblastoma, acute lymphocytic leukemia (ALL) in adults, acute nonlymphocytic leukemia (ANLL) in adults, and indolent non-Hodgkin's lymphoma. MDR1 RNA levels were also increased in some cancers at relapse after chemotherapy, including ALL, ANLL, breast cancer, neuroblastoma, pheochromocytoma, and nodular, poorly differentiated lymphoma. Many types of drug-sensitive and drug-resistant tumors, including NSCLC and melanoma, contained undetectable or low levels of MDR1 RNA. The consistent association of MDR1 expression with several intrinsically resistant cancers and the increased expression of the MDR1 gene in certain cancers with acquired drug resistance indicate that the MDR1 gene contributes to multidrug resistance in many human cancers. Thus, evaluation of MDR1 gene expression may prove to be a valuable tool in the identification of individuals whose cancers are resistant to specific agents. The information may be useful in designing or altering chemotherapeutic protocols in these patients. PMID- 2562855 TI - Structure-activity relationships in prazosin-related compounds. Effect of replacing a piperazine ring with an alkanediamine moiety on alpha 1 adrenoreceptor blocking activity. AB - Several prazosin-related compounds were synthesized in which the piperazine ring of prazosin (1) was replaced by an alkanediamine chain and were evaluated for their blocking activity on alpha 1- and alpha 2-adrenoreceptors in isolated rat vas deferens. All the compounds investigated proved highly selective toward the alpha 1-adrenoreceptor owing to a very low affinity for alpha 2-adrenoreceptors. Furthermore, compounds 2, 9, and 13 were also investigated in vivo to determine their hypotensive effect on anesthetized rats, which were compared with that of prazosin (1). It was confirmed that the piperazine moiety of 1 is not essential for potency. However, optimum activity depends on two parameters: carbon-chain length of the alkanediamine moiety and N-methylation of both the amide and the 2 amino functions. In the desmethyl series, optimum activity was associated with the lower homologues (2-4) bearing a chain of two to four methylenes whereas in the N,N'-dimethyl series peak potency was observed with a six-carbon chain as in 13. Compound 13 proved the most active of the series and was more potent than prazosin (1) in both in vivo and in vitro assays. It is hypothesized that the alpha 1-adrenoreceptor incorporates a lipophilic area that is located between the binding sites for the quinazoline and the furoyl moieties and is able to accommodate a polymethylene chain. PMID- 2562857 TI - Does P-glycoprotein predict response to chemotherapy? PMID- 2562858 TI - Short stature and malnutrition in cystic fibrosis. PMID- 2562859 TI - Topical local anaesthetics and herpes simplex. PMID- 2562860 TI - Parvovirus infection, leukaemia, and immunodeficiency. PMID- 2562861 TI - Protein C deficiency and thrombotic complications after liver transplantation in children. PMID- 2562862 TI - M2 autoantigens in primary biliary cirrhosis. PMID- 2562863 TI - Premalignant epidermal changes due to argon laser. PMID- 2562864 TI - Fund-raising advertisements. PMID- 2562865 TI - Prison medicine. PMID- 2562866 TI - Reporting consensus. PMID- 2562867 TI - Surgical insurrection. PMID- 2562868 TI - Alcohol and the U-shaped curve. PMID- 2562870 TI - Trends in unexpected deaths in Sheffield. PMID- 2562869 TI - Edinburgh declaration and medical education. PMID- 2562871 TI - Bovine pestivirus and human infection. PMID- 2562872 TI - Diagnosis of Pneumocystis carinii pneumonia by non-experts. PMID- 2562873 TI - Paediatric HIV infection. PMID- 2562874 TI - Need to confirm HTLV-1 screening assays. PMID- 2562875 TI - Etretinate and pregnancy. PMID- 2562877 TI - Local anaesthesia for cataract surgery. PMID- 2562876 TI - Glucose tolerance in pregnancy. PMID- 2562878 TI - Monitoring equipment and anaesthetic failures. PMID- 2562879 TI - Geographical relation between Alzheimer's disease and aluminum in drinking water. AB - In a survey of eighty-eight county districts within England and Wales, rates of Alzheimer's disease in people under the age of 70 years were estimated from the records of the computerised tomographic (CT) scanning units that served these districts. Rates were adjusted to compensate for differences in distance from the nearest CT scanning unit and for differences in the size of the population served by the units. Aluminium concentrations in water over the past 10 years were obtained from water authorities and water companies. The risk of Alzheimer's disease was 1.5 times higher in districts where the mean aluminium concentration exceeded 0.11 mg/l than in districts where concentrations were less than 0.01 mg/l. There was no evidence of a relation between other causes of dementia, or epilepsy, and aluminium concentrations in water. PMID- 2562880 TI - Cold-induced pulmonary oedema in scuba divers and swimmers and subsequent development of hypertension. AB - The effect of cold and/or a raised partial pressure of oxygen was examined in eleven people with no demonstrable cardiac abnormality but who had pulmonary oedema when scuba diving or surface swimming, and in ten normal divers. These stimuli induced pathological vasoconstriction in the pulmonary oedema group, nine of whom also showed signs of cardiac decompensation when so stimulated. The pulmonary oedema patients have been followed-up for an average of 8 years. Seven have become hypertensive. Except for the onset of lone atrial fibrillation in one normotensive female diver and development of Raynaud's phenomenon in a normotensive man, there have been no cardiovascular events and no deaths. PMID- 2562881 TI - DNA probes demonstrate a single highly conserved strain of Mycobacterium avium infecting AIDS patients. AB - Strains of the Mycobacterium avium intracellulare complex (MAIC) have become important colonisers of patients with acquired immunodeficiency syndrome (AIDS). Restriction fragment length polymorphisms were used to study the DNA from 88 MAIC isolates, including 51 derived from 47 AIDS patients. MAIC isolates from 33 of 45 AIDS patients were identical at the molecular level and distinct from the mycobacteria isolated from the stools of healthy subjects. The study also showed that serotyping correlates poorly with the genetic identity of these organisms. Mycobacterium paratuberculosis, which has been implicated in Crohn's disease, was not identified in any of the cultures studied. PMID- 2562882 TI - Donor blood frozen and stored between -20 degrees C and -25 degrees C with 35-day post-thaw shelf life. AB - With standard blood bank facilities and interconnected plastic bags donor erythrocytes can be frozen and stored in commercial freezers at temperatures of 20 degrees to -25 degrees C for at least 6 months. The cryoprotectant, a mixture of glycerol and dextrose, is washed off in a closed-circuit system. After being thawed and reconstituted with an additive solution, the pack of red cells can be stored in liquid form for up to 35 days at 4 degrees-6 degrees C. The system lends itself to routine clinical blood transfusion practice. PMID- 2562883 TI - Motoneuron disease: a disorder secondary to solvent exposure? AB - There seems to be a statistically significant association between work in the leather industry and subsequent development of motoneuron disease. The reason for this association may be occupational exposure to solvents, which may damage motoneurons either directly or through activation of latent virus. PMID- 2562884 TI - Genetics of schizophrenia. PMID- 2562885 TI - Adjuvant systemic treatment for breast cancer meta-analysed. PMID- 2562886 TI - Aluminium and Alzheimer's disease. PMID- 2562887 TI - Is cervical laser therapy painful? PMID- 2562888 TI - Listeriosis. PMID- 2562889 TI - Cancer risks of oral contraception. PMID- 2562890 TI - High prevalence of gynaecological diseases in rural Indian women. AB - A population-based cross-sectional study of gynaecological and sexual diseases in rural women was done in two Indian villages. Of 650 women who were studied, 55% had gynaecological complaints and 45% were symptom-free. 92% of all women were found to have one or more gynaecological or sexual diseases, and the average number of these diseases per woman was 3.6. Infections of the genital tract contributed half of this morbidity. Only 8% of the women had undergone gynaecological examination and treatment in the past. There was an association between presence of gynaecological diseases and use of female methods of contraception, but this could explain only a small fraction of the morbidity. In the rural areas of developing countries, gynaecological and sexual care should be part of primary health care. PMID- 2562891 TI - Patients' preference in indomethacin trials: an overview. AB - Meta-analysis was used to study patients' preference in 37 crossover trials that compared indomethacin with newer non-steroidal, antiinflammatory drugs (NSAIDs). 3 reports did not present numerical data. Patients who withdrew from the trial were included in the analysis. The difference between the proportion of patients who preferred the new drug and the proportion who preferred indomethacin (the therapeutic gain) was 14%. After exclusion of 2 unreliable studies the therapeutic gain was only 7%, and when 4 preliminary reports were also ignored, the gain was 5% (95% confidence interval 0 to 10%). In two additional analyses in which the 2 outlying results were excluded, the gain was also 5%. The findings do not support the trend to replace indomethacin with newer NSAIDs. PMID- 2562892 TI - Oesophageal surgery under Akiyama. PMID- 2562893 TI - Demedicalisation of oppression. PMID- 2562894 TI - United States. Cancer risk from domestic radon. PMID- 2562895 TI - Infective cause of childhood leukaemia. PMID- 2562896 TI - Comparing the efficacy of pertussis vaccines. PMID- 2562897 TI - Treatment of essential thrombocythaemia with interferon alpha-2b. PMID- 2562898 TI - Non-invasive sampling method for detecting Chlamydia trachomatis. PMID- 2562899 TI - Treatment for irradiation-induced mucositis. PMID- 2562900 TI - Orchidopexy at or after seven years of age. PMID- 2562901 TI - Misoprostol and ulcer prophylaxis. PMID- 2562902 TI - Aluminium chelator (transferrin) reverses biochemical deficiency in Alzheimer brain preparations. PMID- 2562903 TI - Salmonella enteritidis. PMID- 2562904 TI - Usher syndrome: clinical findings and gene localization studies. AB - The issue of genetic heterogeneity is a critical problem in the localization of the gene(s) for Usher syndrome. Based on the data obtained on families studied to date, the differences between type I and type II Usher syndrome appear quite distinct with regard to auditory and vestibular function. Although the majority of families can be confidently diagnosed as typical type I or type II, clinical investigations revealed four families with findings that did not fit into either of the two more common subtypes. These findings emphasize the critical importance of an in-depth clinical analysis concomitant with the linkage investigation to assure accurate subtyping of Usher syndrome. Based on an analysis of only those families with definite type I or type II Usher syndrome, approximately 17% of the genome can be excluded as a potential site of the gene for type I, and 14% can be excluded as the site for the type II gene. This study will continue until the Usher gene(s) is successfully localized. PMID- 2562905 TI - Antigen presentation by B cells. PMID- 2562906 TI - Chemical codes for the control of behaviour in arthropods. AB - Neuromodulators and hormones elicit and modify well-defined behaviours. Their mode of action can be studied to advantage in arthropods, where the natural releasing cells and neuronal target circuits are concisely identified. The coordinated actions of biogenic amines and peptides on both central and peripheral neural activity and metabolic processes bias the whole organism to perform a coherent behavioural routine. PMID- 2562909 TI - Vecuronium infusion requirements in pediatric patients during fentanyl-N2O-O2 anesthesia. AB - Eighty-one pediatric patients, ranging from neonates to adolescents, were studied during fentanyl-N2O-O2 anesthesia to determine for each of them the vecuronium infusion required to maintain 90-95% neuromuscular block (NMB). Electromyographic monitoring with train-of-four stimuli was used. The steady infusion rate was 62 +/- 15 (SD) micrograms.kg-1.hr-1 in neonates and infants. This rate was 40% of that required by children 3 to 10 years old (154 +/- 49 micrograms.kg-1.hr-1; P less than 0.05). In adolescents the vecuronium requirement was less than in children and was comparable to that reported in adults in other studies (89 +/- 13 micrograms.kg-1.hr-1). Despite considerable individual variation, the infusion rate could be reliably estimated on the basis of duration of greater than 90% NMB maintained by small doses of vecuronium given after intubation. Also, a close correlation existed between the duration of greater than 90% NMB maintained by 100 micrograms/kg of vecuronium and the individual infusion rate (r2 = 0.76). PMID- 2562907 TI - GroE heat-shock proteins promote assembly of foreign prokaryotic ribulose bisphosphate carboxylase oligomers in Escherichia coli. AB - Assembly of foreign prokaryotic ribulose bisphosphate carboxylases (Rubiscos) in Escherichia coli requires both heat-shock proteins groEL and groES. GroEL is related to a chloroplast protein implicated in Rubisco assembly. Bacteria and chloroplasts therefore have a conserved mechanism that uses auxiliary proteins to assist in the assembly of Rubisco. PMID- 2562908 TI - Effects of the tumour promoter okadaic acid on intracellular protein phosphorylation and metabolism. AB - Okadaic acid is a polyether derivative of 38-carbon fatty acid, and is implicated as the causative agent of diarrhetic shellfish poisoning. It is a potent tumour promoter that is not an activator of protein kinase C, but is a powerful inhibitor of protein phosphatases-1 and -2A (PP1 and PP2A) in vitro. We report here that okadaic acid rapidly stimulates protein phosphorylation in intact cells, and behaves like a specific protein phosphatase inhibitor in a variety of metabolic processes. Our results indicate that PP1 and PP2A are the dominant protein phosphatases acting on a wide range of phosphoproteins in vivo. We also find that okadaic acid mimics the effect of insulin on glucose transport in adipocytes, which suggests that this process is stimulated by a serine/threonine phosphorylation event. PMID- 2562910 TI - Ketamine potentiates nondepolarizing neuromuscular relaxants in a primate. AB - Ketamine has many neuromuscular effects in vitro. Its neuromuscular effects in vivo have been controversial and inconsistent. To systematically examine its neuromuscular effects over a wide dose range and its interaction with all popular nondepolarizing neuromuscular relaxants, the effects of ketamine 2, 5, and 10 mg/kg IV were studied on a continuous but incomplete (50%) neuromuscular block preestablished by an IV infusion of d-tubocurarine, atracurium, vecuronium, and pancuronium. Indirectly stimulated adductor pollicis muscle response of monkeys anesthetized with 0.5-1.0% halothane in oxygen were quantified. Ketamine in the absence of a neuromuscular relaxant had no effect on the thumb twitch. In a dose dependent manner, ketamine significantly enhanced the 50% depression of the thumb twitch preestablished by a constant IV infusion of each of the four muscle relaxants studied. Ketamine 2 mg/kg potentiated the neuromuscular relaxants in the following order of magnitude: vecuronium greater than atracurium greater than d-tubocurarine greater than pancuronium. However, with a 10 mg/kg dose of ketamine, pancuronium became as potentiated as was vecuronium, i.e., pancuronium = vecuronium greater than atracurium greater than d-tubocurarine. It is concluded that in the primate, ketamine potentiates all nondepolarizing muscle relaxants in a dose-dependent manner. PMID- 2562911 TI - Negative inotropic effects of esmolol. PMID- 2562912 TI - Intrinsic sympathomimetic activity counteracts beta-blocker inhibition of renin activation. AB - Enzymatically inactive renin (IR) is the predominant circulating form of renin. Sympathetic activity may influence plasma renin activity (PRA) by regulation of the conversion of IR to active renin (AR, PRA). It has been demonstrated previously that beta blockade lowers PRA at least partly through inhibition of this conversion process. The authors hypothesized that beta blockade and intrinsic sympathomimetic activity (ISA) would have opposing effects on production of AR from its inactive precursor. Eighteen primary hypertensives (12 male, 6 female, mean age 57.7 +/- 2.7) were entered in a placebo-controlled, double-blind crossover study of the effects of equipotent doses of pindolol and propranolol on mean +/- SEM systolic BP, diastolic BP, heart rate, active renin (AR), total renin (TR), inactive renin (IR), and % AR/TR. Drug dose was titrated to achieve a goal DBP of 90 mmHg or less. Active renin was defined as the rate of generation of angiotensin I in 37 degrees C plasma at pH 5.7. Total renin was determined by preincubation of plasma aliquots with 1.5 mg/mL trypsin in the presence of 5 mM benzamadine for one hour at -4 degrees C prior to assay of renin activity. Inactive renin was calculated as TR minus AR. The BP responses achieve by dose titration of propranolol and pindolol were virtually identical at rest, indicating equivalent depressor effects of the two beta blockers. Heart rate and active renin were, however, lowered to a much greater extent with propranolol as compared with pindolol. The lack of significant pindolol-induced fall in % AR/TR suggests that this drug has little net effect on the formation of AR from IR.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562913 TI - Superiority of activated charcoal alone compared with ipecac and activated charcoal in the treatment of acute toxic ingestions. AB - A prospective, randomized clinical trial compared the clinical effectiveness of syrup of ipecac and activated charcoal to that of activated charcoal alone in the treatment of acute toxic ingestions. Two hundred adult patients with mild to moderate oral overdoses were entered into the trial. Patients receiving only activated charcoal were discharged from the emergency department in significantly (P less than or equal to .05) less time than those receiving both syrup of ipecac and activated charcoal (6.0 +/- 0.3 vs 6.8 +/- 0.2 hours, respectively). The percentage of patients requiring nonpsychiatric hospitalizations was not significantly different between the two groups (11.2% vs 14.0%, respectively). For the hospitalized patients, the length of time spent in the ICU and in the hospital was not statistically different between the two groups. A complication rate of 5.4% was found with the ipecac and activated charcoal treatment compared with a 0.9% complication rate in the activated charcoal group (P less than or equal to .05). Three episodes of aspiration pneumonitis occurred after administration of ipecac and activated charcoal, while no episodes of aspiration were noted after treatment with only activated charcoal. Together, these data are consistent with the recommendation that ED treatment with activated charcoal alone be the gastrointestinal decontamination procedure of choice for the routine mildly-to-moderately orally poisoned adult patient. PMID- 2562914 TI - Neuroendocrine and mood responses to intravenous L-tryptophan in 3,4 methylenedioxymethamphetamine (MDMA) users. Preliminary observations. AB - 3,4-Methylenedioxymethamphetamine (MDMA; "ecstasy") is a selective serotonin (5 HT) neurotoxin in laboratory animals. To assess its effects on 5-HT function in humans, serum prolactin (PRL) and mood responses to intravenous L-tryptophan were measured in nine recreational users of MDMA and compared with findings from nine matched healthy controls. L-Tryptophan induced a rise in the PRL concentration in controls, but not in MDMA users. Peak change and the area under the curve of the PRL response appeared to be blunted in MDMA users, but the difference from controls did not reach statistical significance. This study provides suggestive evidence of altered 5-HT function in MDMA users, but more definitive studies clearly are needed. PMID- 2562915 TI - Major depressive disorder and immunity. Role of age, sex, severity, and hospitalization. AB - An association between depression and altered immunity has been suggested but has not been consistently demonstrated. We have studied 91 patients with unipolar major depressive disorder, and no mean differences were found between the patients and concurrently studied matched controls in mitogen-induced lymphocyte proliferation, lymphocyte subsets, and natural killer cell activity. There were, however, significant age-related differences between the depressed patients and controls in mitogen responses and in the number of T4 lymphocytes. In contrast to age-related increases in mitogen response and in T4 cells in controls, depressed patients did not show increased lymphocyte responses or numbers of T4 lymphocytes with advancing age. Severity of depression and hospitalization status were also associated with immune system changes. Altered immunity does not appear to be a specific biologic correlate of major depressive disorder but may occur in subgroups of depressed patients. PMID- 2562916 TI - Predictors of drug response in depression. AB - Although the major classes of antidepressant drugs have been available for over 30 years, clinicians are still unable to predict accurately the response of their depressed patients to medication. This article reviews both clinical and biologic predictors of treatment response and makes recommendations for future studies. The tricyclic antidepressants remain the drugs of choice in major depressive disorders. Lithium has a place in bipolar depressions. Monoamine oxidase inhibitors have a role in depressions accompanied by marked anxiety and/or panic symptoms, in patients who have previously responded to them, and as a second choice treatment in those depressed patients who have not responded to tricyclic antidepressants. Electroconvulsive therapy or additional antipsychotic drugs are frequently necessary in very severe and delusional depressions. Biologic predictors of response, despite some interesting leads that may in the long term be of considerable importance, are not yet sufficiently established to be of routine clinical usefulness, although either dexamethasone nonsuppression or a shortened rapid eye movement latency may identify depressed patients who require biologic treatment. PMID- 2562917 TI - Endocrine pancreas in chronic pancreatitis. A qualitative and quantitative study. AB - This study includes nine patients with diabetes mellitus (DM) and chronic pancreatitis (CP) (group I); 11 patients without DM and with CP (group II); and a control group (group III) consisting of five autopsy cases with neither DM nor CP. These groups were evaluated by routine histologic stains and immunocytochemical stains for insulin, glucagon, and somatostatin. Semiquantitative assessment of the degree of exocrine pancreatic atrophy and of two endocrine features (diffuse endocrine proliferation and ductoendocrine proliferation) was performed for each pancreas. Quantitative determination of the cell composition was carried out in three kinds of islets (parenchymal, sclerosis, and newly formed). The mean percentages of the insulin-producing B cells were significantly lower in the parenchymal (44.5%) and new (34.3%) islets of diabetic patients than in the controls (67.8%) and parenchymal (59.4%) islets of nondiabetic patients. The mean percentages of glucagon-producing A cells revealed significant increases in the parenchymal (43.0%) and new (55.7%) islets of diabetic patients as compared with the controls (24.3%) and parenchymal (32.2%) islets of nondiabetic patients. The mean percentage of somatostatin producing D cells was significantly increased in the parenchymal islets (12.4%) of diabetic patients as compared with the parenchymal islets (8.2%) of nondiabetic patients and controls (7.5%). These findings correlate with clinical data of frequent DM in CP, but are partly in contrast with previous immunohistochemical analysis findings in CP. PMID- 2562918 TI - The interaction of temelastine with cytochrome P450 mixed-function oxidase enzymes in vivo and in vitro. PMID- 2562919 TI - Purification and partial characterization of glutamine synthetase from the photosynthetic bacterium Rhodospirillum rubrum. AB - Glutamine synthetase (L-glutamate: ammonia ligase (ADP-forming), EC 6.3.1.2) from the photosynthetic bacterium Rhodospirillum rubrum grown under nitrogen fixing conditions has been purified to homogeneity. The purification procedure involves affinity chromatography on ADP-agarose type 2 as the major purification step. The recovery in the purification is 70%. The specific activity of the purified enzyme is about 10-times higher in the gamma-glutamyl transferase assay than in the coupled biosynthetic assay. The molecular weight was determined to 530,000 by native gradient polyacrylamide gel electrophoresis and to 500,000 by gel filtration. The subunits have an apparent molecular weight of 52,000. Glutamine synthetase isolated from Rsp. rubrum which had been exposed to ammonium ions ('switch-off') before harvest had about 20% of the transferase activity compared with the enzyme purified from nitrogen-starved cells. The low-activity form showed two bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PMID- 2562920 TI - Differential expression of CD11b/CD18 (Mo1) and myeloperoxidase genes during myeloid differentiation. AB - During the course of differentiation of early human myeloid cells toward monocytes and granulocytes, cell surface expression of the cell adhesion molecule, CD11b/CD18 (Mo1) increases dramatically and expression of myeloperoxidase (MPO), a bacteriocidal enzyme, decreases markedly. Using the inducible promyelocytic cell line HL-60 as a model, we studied the mRNA expression of these genes. Differentiation of these cells along both a monocytic and a granulocytic pathway demonstrated that the mRNA levels of the two subunits of CD11b/CD18 increased in a pattern temporally and quantitatively similar to the increase in cell surface expression of this heterodimer. In contrast, the expression of MPO mRNA decreased in a temporal and quantitative pattern similar to the known decrease in MPO protein during differentiation, suggesting that regulation of these myeloid-specific proteins may occur at the level of mRNA expression. These findings have important implications with regard to the nature of the block in differentiation in acute nonlymphocytic leukemia and the regulation of myeloid gene expression. PMID- 2562921 TI - Treatment of aggressive multiple myeloma by high-dose chemotherapy and total body irradiation followed by blood stem cells autologous graft. AB - Eight patients with stage III aggressive multiple myeloma, refractory to current chemotherapy in six cases, were treated by high-dose chemotherapy (nitrosourea, etoposide, and melphalan) (HDC) and total body irradiation (TBI), followed by autografting with blood stem cells. These cells were previously collected by leukapheresis performed during hematologic recovery following cytotoxic drug induced bone marrow aplasia. Seven patients were alive 9 to 17 months after HDC TBI and graft. One died at day 40 from cerebral bleeding. All living patients achieved a 90% or greater reduction in tumor mass. In two cases, a complete remission (CR) has persisted at a follow-up of 15 and 16 months. Three patients have been well and off therapy with stable minimal residual disease (RD) since 10, 11, and 17 months, respectively. A patient in apparent CR and another with RD have relapsed 9 to 12 months posttreatment. Autologous blood-derived hematopoietic stem cells induced successful and sustained engraftment in all living patients. These results, although still preliminary, indicate that HDC and TBI, followed by blood stem cells autograft, which has both practical and theoretical interest over allogeneic or autologous bone marrow transplantation, deserve consideration in selected patients with multiple myeloma. PMID- 2562922 TI - Chromosome 7 long arm deletion in myeloid disorders: a narrow breakpoint region in 7q22 defined by molecular mapping. AB - The involvement of the erythropoietin (EPO), plasminogen activator inhibitor type I (PAI1), and multi-drug resistance (MDR2) genes located in chromosomal region 7q21-22 was studied in patients with myeloid disorders and with or without a chromosome 7 abnormality. Separated blood mononuclear cells and granulocytes from 21 patients were used in restriction fragment length polymorphism (RFLP) studies with gene-specific DNA probes. A marked weakness of one of the allelic bands was observed in granulocyte-derived DNA from heterozygous patients with monosomy 7. In four patients with a partial deletion of chromosome 7 long arm (7q-), marked weakness of an allelic band was observed in granulocyte-derived DNA with PAI1 probe (four heterozygous patients) and MDR2 probe (one heterozygous patient), implying deletion of these genes. In contrast, the EPO gene was not deleted in these patients, as demonstrated by the presence of two allelic bands of equal strength in granulocyte-derived DNA (two patients) or by gene dosage estimation (two patients). Two allelic bands of equal strength were also observed in three heterozygous patients with an arbitrary probe (pKV13) located in 7cen-q21.3. Unexpected hemizygosity or hybridization bands were not observed in any patient. We conclude that PAI1 and MDR2 are located distally of EPO in 7q22, and that none of these genes is commonly rearranged in myeloid disorders. The chromosome 7 long arm deletion breakpoint is located in a relatively narrow segment between the PAI1 and EPO genes in different patients. The deletion may involve a specific site in DNA, since the genetic distance between the PAI1 and EPO genes is only 3 cM. PMID- 2562923 TI - Phenotypic heterogeneity of spontaneous lymphomas of CWD mice. AB - Animals of the inbred mouse strain, CWD, express endogenous murine leukemia viruses early in life and have a high incidence of spontaneous neoplasms. We found that approximately one half of these animals died of malignant lymphoma by the age of 16 months. Splenic enlargement was seen in all mice, but thymic involvement was unusual. One half of the CWD tumors were diffuse lymphoblastic or immunoblastic lymphomas while the remainder were large cell, small cell, or mixed cell lymphomas. Analysis of DNAs from 12 tumors for immunoglobulin and T-cell receptor gene rearrangements revealed that all six of the lymphoblastic and immunoblastic lymphomas were of T-cell origin, as was one tumor of small cleaved cells. Four of the others were clonal B-cell lymphomas and one was of uncertain lineage. Assays of a limited number of tumors for the expression of the Thy 1.2 and IgM molecules confirmed the diversity in the cellular phenotype. The results indicate that CWD mice develop primarily splenic lymphomas with an unusual degree of heterogeneity in the tumor cell phenotypes as compared with the thymic lymphomas found in other high leukemia strains. The CWD strain is a useful new model for studies of retroviral leukemogenesis and the relationship between the histopathology and immunophenotype of malignant lymphomas. PMID- 2562924 TI - Clonal analysis of myelodysplastic syndromes: evidence of multipotent stem cell origin. AB - Restriction fragment length polymorphisms (RFLPs) of the X-chromosome genes hypoxanthine phosphoribosyl transferase (HPRT) and phosphoglycerate kinase (PGK) were studied in 34 female patients with primary myelodysplastic syndromes (MDS). Twelve patients (35%) were heterozygous at the HPRT or PGK loci for BamHI or BglI RFLPs, respectively. In eight patients showing PGK polymorphisms, clonality was determined by X-chromosome inactivation analysis. These included patients from different morphologic subtypes: four with refractory anemia (RA), two with RA and ring sideroblasts (RARS), one patient with RA with excess of blasts (RAEB), and one with chronic myelomonocytic leukemia (CMML). A monoclonal pattern of X chromosome inactivation was observed in seven cases. In a further case characterized by bone marrow hypoplasia, peripheral blood (PB) leukocytes were polyclonal in origin. Following low-dose cytarabine therapy, reversion to polyclonal hematopoiesis was observed in a case of RAEB indicating the presence of residual normal hematopoietic stem cells with the capacity for marrow reconstitution. The clonal relation of lymphoid and granulocyte/monocyte lineages was studied directly in two cases of CMML exhibiting somatic mutations of N-ras or Ki-ras oncogenes. By selective oligonucleotide hybridization to ras gene sequences amplified in vitro by the polymerase chain reaction, a mutated ras allele was demonstrated in PB granulocytes, monocytes, and B and T lymphocytes of both patients. We conclude that MDS arise from a multipotent hematopoietic stem cell with the potential for myeloid and lymphoid differentiation. PMID- 2562925 TI - Laboratory diagnosis of the first cases of HIV-2 infection in Canada. AB - Until recently the geographic distribution of infection due to human immunodeficiency virus type 2 (HIV-2) had excluded North America. We report the first two cases of such infection in Canada. Both people came from endemic areas of western Africa and were asymptomatic. The results of a commercial enzyme immunoassay specific for HIV-1 antibody were positive in both cases, but those of the Western blot technique were indeterminate. The Western blot technique specific for HIV-2 antibody and the indirect fluorescent antibody test were used to verify the presence of HIV-2 antibody. PMID- 2562926 TI - Association of elevated sulfatides and sulfotransferase activities with human renal cell carcinoma. AB - Lactosylceramide sulfate and galactosylceramide sulfate were found to increase markedly in human renal cell carcinoma (adenocarcinoma) as compared to uninvolved tissue. Activities of two sulfotransferases toward galactosylceramide and lactosylceramide as substrates were significantly elevated in the carcinoma compared to the uninvolved tissue resulting in enhanced synthesis of the two sulfatides in the carcinoma. The elevation of the two sulfotransferases was parallel in most tumors, suggesting that the same enzyme is responsible for the enhanced synthesis of two sulfatides. No consistent difference in the activity of arylsulfatase A, which desulfates the two sulfatides, was observed between the carcinoma and uninvolved tissue. Both the present and previous results show that the increased synthesis of the sulfatide(s) due to elevated sulfotransferase activity could be a biochemical characteristic common to adenocarcinomas derived from different tissues. PMID- 2562927 TI - Metabolic studies of estrogen- and tamoxifen-treated human breast cancer cells by nuclear magnetic resonance spectroscopy. AB - The effects of 17 beta-estradiol treatment versus tamoxifen on the metabolism of human breast cancer T47D-clone 11 cells were studied by noninvasive 31P and 13C nuclear magnetic resonance techniques. 31P nuclear magnetic resonance spectra revealed differences between estrogen and tamoxifen treated cells. The steady state content of phosphorylcholine and of the nucleoside diphosphates was higher in the tamoxifen treated cells by 33 and 140%, respectively, relative to estrogen treated cells. The intracellular pH of 7.2 and the content of the nucleoside triphosphates, Pi, phosphocreatine, glycerolphosphorylcholine, and glycerolphosphorylethanolamine and uridine diphosphoglucose remained the same in both treatments. Glucose utilization and subsequent lactate, glutamate, alanine, and glycerol 3-phosphate synthesis were monitored on line following administration of specifically labeled [13C]glucose. In estrogen treated cells the rate of lactate production via glycolysis was 560 fmol/cell/h and the initial rate of 13C labeling of the glutamate pool via the Krebs cycle was 6.8 fmol/cell/h. In the tamoxifen treated cells these rates were 2-fold lower, at 250 and 2.9 fmol/cell/h for lactate and glutamate labeling, respectively. In estrogen treated cells, the calculated content of glutamate (19 fmol/cell), alanine (11 fmol/cell), and glycerol 3-phosphate (8 fmol/cell) was higher than in tamoxifen treated cells, where only glutamate labeling was detected (13 fmol/cell). The observed differences in the in vivo kinetics of glucose metabolism may provide a sensitive measure for detecting the response of human breast cancer cells to estrogen versus tamoxifen treatments. PMID- 2562928 TI - Vascular endothelial cells enhance T cell responses by markedly augmenting IL-2 concentrations. AB - Cultured human endothelial cells (EC), dermal fibroblasts (DF), and blood monocytes were compared for their effects on IL-2 concentration measured in the medium of both unfractionated peripheral blood mononuclear cells (PBMC) and highly enriched CD4+ T cell populations maximally stimulated by the polyclonal mitogen phytohemagglutinin (PHA). EC, but not DF or blood monocytes, could markedly augment IL-2 concentrations, up to 30-fold or more. This action of EC could not be replaced by fixed EC, EC-conditioned medium, or recombinant IL-1. Antibody to CD2 but not to CD18 blocked the EC effect. The augmented IL-2 concentrations generated in the presence of EC appeared biologically active in that the addition of living EC conferred a proliferative advantage to PBMC at suboptimal PHA concentrations, an effect which could be mimicked by exogenous IL 2. We propose that EC augmentation of IL-2 synthesis may contribute to the relatively unique ability of EC to stimulate a primary allogeneic response in vitro and may function in vivo to boost T cell responses to limiting quantities of antigen. PMID- 2562929 TI - A previously unrecognized large fraction of cytotoxic lymphocyte precursors is present in CD4+ human peripheral blood T cells. AB - We describe a limiting dilution (LD) culture system in which cell sorter-purified CD4+ (and CD8+) peripheral blood T cells are cocultured with irradiated, anti-CD3 mab-producing OKT3 hybridoma cells. Under these conditions, one out of 2-3 CD4+ (and CD8+) T cells is induced to clonal proliferation. In striking contrast to previously described LD culture systems, every growing CD4+ cell clone displayed cytotoxic activity when tested in a lectin-facilitated 51Cr release assay against P815 target cells. This contrasts with the development of cytotoxic CD4+ T cells in alloantigen-stimulated LD cultures, where only one out of 15-20 proliferating CD4+ cells killed P815 in the presence of PHA, and one out of 300-500 proliferating CD4+ cells displayed alloantigen-specific cytotoxic activity. Furthermore, we have established antigen-specific proliferating CD4+ T cell clones which do not exert antigen-specific cytotoxicity but can be cytotoxic when crosslinked to target cells via lectin or monoclonal antibody (anti-CD3, anti TCR). Our results show that a previously unrecognized large fraction (at least 30 50%) of all peripheral blood CD4+ T cells can give rise to cytotoxic effector cells. The mode of CD4+ T cell activation (OKT3 hybridoma versus alloantigen) thus determines whether the intrinsic cytotoxic capacity of CD4+ T cells is functionally activated or not. PMID- 2562930 TI - Role of different lymphocyte subsets in human anti-viral T cell cultures. AB - We have systematically studied uncloned human cell lines derived from anti influenza A virus or anti-Epstein-Barr virus (EBV) bulk cultures, or from cultures highly enriched for CD4+ or CD8+ lymphocytes. The most noteworthy results are the following: (1) Anti-viral bulk cultures consisted of more than 90% of CD8+ cells in all cases. In contrast, anti-HLA cell lines are composed of approximately 50% CD8+ and 50% CD4+ cells. All of the CD8+ and CD4+ cells present in the culture were also 4B4+/2H4-. (2) In anti-viral bulk cultures, the cytolytic activity was restricted by HLA class I molecules and almost exclusively through a single HLA class I molecule. (3) Positively or negatively selected CD8+ lines showed the same restriction pattern. They grew less efficiently than bulk cultures but could be maintained in the absence of CD4+ cells. The CD4+ cells were however necessary at the beginning of the culture for the development of cytolytic anti-influenza virus CD8+ cells, whereas they were not required for the development of cytolytic anti-EBV CD8+ cells. (4) The CD4+ cell lines grew more actively than bulk cultures. A cytolytic activity for virus-infected cells was constantly detected in these culture from the third passage onward and it was always restricted by HLA class II molecules. This activity was maintained throughout the culture period. However, class II-restricted cytolytic cells were not detected during primary or secondary responses in vitro. PMID- 2562931 TI - Increased proportion of CD4+CDw29+CD45R-UCHL-1+ lymphocytes in the cerebrospinal fluid of both multiple sclerosis patients and healthy individuals. AB - In this study we confirm earlier reports of an increase of the proportion of T and CD4+ lymphocytes and a decrease of B and CD8+ lymphocytes in cerebrospinal fluid (CSF) as compared to peripheral blood (PB) in MS patients. In addition we now demonstrate that this difference between CSF and PB lymphocyte populations is of the same magnitude in healthy individuals suggesting that it is physiological and not associated with disease. Functionally distinct subsets of the T human helper cell (CD4+) population have previously been defined by the monoclonal antibodies 4B4 (CDw29), Leu-18 (CD45R), and UCHL-1. In the present investigation we demonstrate a selective increase in the proportion of CD4+CDw29+CD45R-UCHL-1+ lymphocytes in CSF as compared to PB of both MS patients and healthy individuals, which strongly indicates that also this enrichment is physiological rather than associated with disease. A possible relationship between this subset of CD4+ lymphocytes and T memory cells is discussed. PMID- 2562932 TI - Regulation of in vitro PWM-induced IgG secretion in humans. AB - PWM-induced differentiation of human PBMC into immunoglobulin (Ig) secreting cells is a widely used model of in vivo IgG secretion. We examined the relationship between the amount of IgG secreted in PBMC cultures obtained from individuals who consistently secrete either very high (HR) or very low amounts (LR) of IgG and various in vitro immune function assays. The PBMC obtained from LR contained a higher ratio of cells expressing the T-suppressor/inducer to T helper/inducer phenotype. The autologous mixed lymphocyte response was lower and the amount of in vivo radiation sensitive suppression was higher in the LR than in the HR. LR E- cells secreted less IgG than the HR E- cells when both were mixed with heterologous HR E+ cells. Monocyte depletion reduced IgG secretion in both LR and HR. These results suggest that each of the subsets Ts, Th (Thi, Tsi), and B lymphocytes are involved in the regulation of PWM-induced Ig secretion and that the functions of each of these subsets differ in HR and LR individuals. PMID- 2562933 TI - Selective lysis of target cells by interleukin-2-expanded peripheral blood mononuclear leukocyte clones. AB - The presence of distinct cytolytic subsets within interleukin-2-expanded peripheral blood leukocytes (IEL) cultures was demonstrated by clonal analysis. Thirty-seven IEL clones were isolated from two healthy blood donors; 21 destroyed both Daudi and K562 cell lines. Of those 21 clones, 1 clone could destroy autologous PBM, 7 clones could destroy fresh allogeneic ovarian carcinoma (OVA CA) cells, and 6 clones could destroy normal autologous PBM and fresh OVA-CA cells. Twelve of the 37 clones destroyed only one of the four targets tested: 8 clones destroyed K562, 2 clones destroyed Daudi, and 1 clone each was selective for autologous PBM or OVA-CA. Of the remaining 4 clones, 1 destroyed OVA-CA and Daudi cells, 1 destroyed PBM and K562, 1 destroyed PBM and Daudi cells, and 1 destroyed PBM, Daudi, and OVA-CA. These results suggest that these functionally heterogeneous cytolytic clones may use different cell recognition or cytolytic mechanisms to enable these distinct and, at times, reciprocal patterns of target cell selectivity. PMID- 2562934 TI - Persistence of the irradiated host component in thymocyte populations from bone marrow radiation chimeras infected with lymphocytic choriomeningitis virus. AB - The thymus of chimeras made using T cell-depleted donor bone marrow from Thy1.1+ mice and 950 rad Thy 1.2+ recipients is dominated initially by cells expressing the Thy 1.2+ phenotype of the irradiated host. The thymocyte population recovered at 2 weeks after reconstitution comprises 80% Thy 1.2+ cells (host), the remainder being Thy 1.1+ (donor). This situation is normally reversed within a further week, with the host Ty 1.2+ (donor). This situation is normally reversed within a further week, with the host Thy 1.2+ thymocytes being present at a frequency of less than 5% from Week 4. Infection with lymphocytic choriomeningitis virus (LCMV) at 1 week after reconstitution with bone marrow causes a profound and persistent drop in the total number of thymocytes. The decline is equivalent for all categories of donor-derived thymocytes defined by two-color flow microfluorometric analysis for CD4 and CD8. However, there is a partial compensation by the retention of cells originating from the Thy 1.2+ host, which constitute 30-40% of the total thymocyte pool as late as 8 weeks after administration of bone marrow in the LCMV-infected chimeras. These radiation-resistant precursors give rise to CD4-8-, CD4-8+, CD4+8-, and CD4+8+ thymocytes, with the latter category being present at increased frequency. The potential skewing of the mature T cell repertoire as a consequence of persistent virus infection is discussed. PMID- 2562935 TI - Flow cytometric analysis of human lymphocytes using affinity-purified antibody to T cell receptor beta synthetic J region peptide. AB - The purpose of this study was to use affinity-purified polyclonal antibodies produced against a synthetic peptide corresponding to the joining (J) region of a human T cell receptor beta chain to characterize antigen receptor expression on subpopulations of human lymphocytes. The synthetic peptide used was ANYGYTFGSGTRLTVV, corresponding to the J segment of the human beta-chain gene YT35. Biochemical characterization has previously demonstrated binding of anti-J beta peptide antibodies to the alpha/beta heterodimer and to certain immunoglobulin light chains. Flow cytometric analysis of normal human peripheral blood lymphocytes performed here, using affinity-purified antibodies to the J beta peptide, showed expression of the epitope on 50-60% of CD20 (B1)-positive B lymphocytes, and on 40-50% of CD8-positive T lymphocytes. Only background levels were observed on CD4-positive T cells. PMID- 2562936 TI - Role of beta 1-receptors and vagal tone in cardiac inotropic and chronotropic responses to a beta 2-agonist in humans. AB - To assess the contribution of cardiac beta 2-receptors in the cardiac inotropic and chronotropic responses to a beta 2 agonist, terbutaline was infused (0.2 and 0.4 micrograms/kg/min), alone or after pretreatment with either oral atenolol 50 mg or atropine 0.04 mg/kg i.v. or both in six healthy subjects with a multiple crossover design. Terbutaline 0.2 micrograms/kg/min increased heart rate by 15 +/ 2 beats/min, and this response doubled (to 29 +/- 3 beats/min) when the terbutaline infusion followed atropine pretreatment, whereas atenolol pretreatment had no significant effect. Heart rate increased by 44 +/- 2 beats/min in response to terbutaline 0.4 micrograms/kg/min. This response was not affected by atropine. Pretreatment with atenolol diminished the chronotropic response to the higher dose of terbutaline to 27 +/- 4 beats/min. The inotropic response (i.e., changes in pressure: volume ratio) to terbutaline 0.2 micrograms/kg/min was potentiated by atropine (from 1.6 +/- 0.3 to 3.4 +/- 0.8 mm Hg/ml), whereas atenolol pretreatment had no effect. At the higher dose of terbutaline, atropine pretreatment had no additional effect, whereas atenolol decreased the rise in pressure: volume ratio from 6.0 +/- 1.4 to 2.6 +/- 1.0 mm Hg/ml. The results with atenolol pretreatment indicate that cardiac beta 1 responses are associated with the higher dose of terbutaline, either through direct beta 1 stimulation or indirectly from presynaptic beta 2 activity. The atropine data show that vagal tone actually increased during the terbutaline infusions, blunting the cardiac effects. The results of the present study support the existence of functional chronotropic, as well as inotropic, beta 2 receptors in the healthy human heart. PMID- 2562937 TI - Nitrate tolerance in epicardial arteries or in the venous system is not reversed by N-acetylcysteine in vivo, but tolerance-independent interactions exist. AB - N-acetylcysteine is assumed to reverse nitrate tolerance by replenishing depleted intracellular sulfhydryl groups, but data on interactions of N-acetylcysteine and nitrates in patients with stable angina are controversial and disappointing. Therefore, we studied the effect of N-acetylcysteine on nitrate responsiveness of epicardial arteries and of the venous system (assessed as changes in effective vascular compliance) in dogs (n = 12) during long-term nitroglycerin treatment (1.5 micrograms/kg/min i.v. for 5-6 days). In dogs with nitroglycerin-specific tolerance (shift of venous or epicardial artery dilation to 15-17-fold higher dosages), N-acetylcysteine (100 mg/kg i.v.) had no dilator effect and did not alter the dose-response relations of nitroglycerin. Yet, in nontolerant dogs (n = 17), N-acetylcysteine augmented (1.5-2.0-fold) the dilation of epicardial arteries and the reduction of peripheral vascular resistance induced by 0.5-1.5 micrograms/kg/min nitroglycerin. In vitro, the augmentation of purified guanylate cyclase activity by nitroglycerin (10-100 microM) was potentiated by N acetylcysteine (0.01-1.0 mM) in saline or in canine plasma, but N-acetylcysteine alone was ineffective. We conclude that 1) N-acetylcysteine does not restore nitroglycerin responsiveness in tolerant epicardial arteries or veins in vivo, 2) a small, tolerance-independent augmentation of nitroglycerin-induced dilation may result from N-acetylcysteine-induced extracellular formation of a stimulant of guanylate cyclase from nitroglycerin. PMID- 2562938 TI - Therapeutic options in the management of chronic heart failure. Is there a drug of first choice? PMID- 2562939 TI - Recent advances in bone marrow transplantation. AB - An increasing number of diseases may be treated successfully by allogeneic bone marrow transplantation (BMT). Initially used for the treatment of immunodeficiency where a cell series or product is replaced, it has now become routine treatment for many forms of leukemia where the transplant provides the rescue after lethal marrow ablation. Recently, diseases such as thalassemia and other inherited metabolic diseases have also been treated by BMT. Formerly the problems of BMT were mainly concerned with graft versus host disease (GVHD) in HLA-matched transplants with HLA-mismatched ones not being possible as GVHD was usually fatal. Since the development of techniques for T cell removal the incidence of GVHD has greatly diminished. T cell removal has also allowed HLA haploidentical mismatched grafts to be performed successfully for immunodeficiency, but there is still a high graft rejection rate in leukemia. This also occurs to a lesser extent with HLA-matched grafts in leukemia. Furthermore, in certain forms of leukemia, particularly chronic granulocytic leukemia, the relapse rate after T cell-depleted BMT is much higher. Trials of better forms of bone marrow conditioning of the recipient are being attempted in order to prevent graft rejection and leukemia relapse. These include total lymphoid irradiation, heavier irradiation and chemotherapeutic regimens, or the use of in vivo monoclonal antibodies such as CAMPATH 1G or anti-LFA-1 (CD11a). In the future, positive selection of stem cells combined with hemopoietic growth factors may allow engraftment without graft versus host disease. This should become the method of choice for autologous transplantation for malignancy. Two monoclonal antibodies directed against the human progenitor cell antigen 1 (HPCA 1) (CD34) have been used for autologous positive stem cell selection in primates and these cells gave full hemopoietic reconstitution in the animals following lethal total body irradiation. PMID- 2562940 TI - Role of interleukins in induction and regulation of human IgE synthesis. AB - Studies of human IgE synthesis are summarized and provide further insight into the cellular and molecular mechanisms involved in IgE regulation, as well as in the alterations responsible for IgE disregulation in some pathological conditions. These include the demonstration that IL-4 is the essential factor for the induction of human IgE syntheses. Another T cell-derived lymphokine, IFN gamma negatively regulated the IgE synthesis induced by IL-4. These two lymphokines can be produced by different T helper cells, as shown in mice, but they can also be the product of the same T cells clones. Additional cellular and/or molecular signals appear to be involved in the IL-4-induced IgE synthesis, but their precise role in this process is undetermined. Finally, alternations of one or more of these regulatory mechanisms can be detected in patients with pathological conditions characterized by hyperproduction of IgE. In particular, the increased prevalence of T cells clones able to produce IL-4 appears to be a distinctive feature of patients with common atopy whereas a reduction in the proportion of IFN-gamma-producing T cells seems to be peculiar of both patients with hyper-IgE syndrome and patients with AIDS. PMID- 2562941 TI - Immunopathogenic mechanisms of HIV infection. AB - The acquired immunodeficiency syndrome (AIDS) follows infection with the human immunodeficiency virus (HIV) after a long and variable period of time. Although HIV can be rapidly cytopathic in vitro for T4 cells, during early in vivo infection, T4 cell numbers are normal and few infected T4 cells can be found. There is increasing evidence that cells of the monocyte/macrophage lineage, including bone marrow precursor cells, can be infected by HIV and are relatively resistant to the cytopathic effects of the virus. As in other lentivirus infections, the monocyte/macrophage may serve as a reservoir of HIV in the body and may play a major role in the pathogenesis of AIDS. In vitro induction of HIV from a latent or low level chronic infection to an active state results from exposure to physiologic cellular inductive signals that might be encountered during normal immune responses. PMID- 2562942 TI - Clinical and immunological aspects of HIV infection in drug addicts. AB - Intravenous drug users (IVDUs) account for more than 64% of the total AIDS cases in Italy. The IVDUs' seropositivity rate is greater than 70% in Milan and greater than 50% in the main cities of Italy. The first evidence of seropositivity in this population dates back to 1979. In a cohort study performed in Milan the rate of progression to overt AIDS among IVDUs was 6% in 3 years (1984-1987). At presentation, more than 75% of the subjects had CD4+ cell counts higher than 400/mm3 (mean 631, median 528, mode 465). These values are significantly higher than those observed in the same population in New York, the only American city with HIV-infection spread comparable to that observed in Milan. The probability of having CD4+ cell counts lower than 400, 300, and 200/mm3 in relation to the length of follow-up was, respectively, 50, 40, and 2% after 36 months from presentation. At the same end point, subjects presenting less than 400 CD4+ cells at entry had 30% probability of falling under 200 cell/mm3. The pattern of CD4+ cells, as much as the low percentage of yearly progression to overt AIDS, is probably related to the recent, even if rapid, spread of infection among IVDUs in Italy. The clinical features of overt AIDS present some differences between IVDUs and other at-risk groups. Among U.S. IVDUs with AIDS, Kaposi's sarcoma is infrequent (2.9% vs 27.7% in homosexual men) while mycotic infections such as deep candidiasis and cryptococcosis are significantly more frequent. The same pattern has been observed in our case file in Milan: esophageal candidiasis represents the most frequent cause of diagnosis of overt AIDS. Mycotic infections, overall, are more frequent than in U.S. IVDUs. The increased rate of mycotic infections among IVDUs might be justified by altered functions of nonspecific immunity, such as PMNL killing and phagocytosis of Candida albicans spores, impaired in HIV-infected IVDUs, but generally normal in infected subjects belonging to the other at-risk groups. PMID- 2562943 TI - Decreased myometrial beta-adrenoceptors in women receiving beta 2-adrenergic tocolytic therapy: correlation with lymphocyte beta-adrenoceptors. AB - We have determined simultaneously the density of beta-adrenoceptors in human myometria (by (-)-[125I]iodopindolol binding) derived from 36 women undergoing cesarean section and in the corresponding circulating lymphocytes (by (-) [125I]iodocyanopindolol binding). In myometrial membranes about 80% to 85% of the beta-adrenoceptors were of the beta 2-subtype. The density of myometrial and lymphocyte beta-adrenoceptors in women treated with the beta 2-adrenoceptor agonist hexoprenaline to prevent preterm labor was about 65% to 70% lower than that in nontreated women. Concomitantly, in hexoprenaline-treated women the 10 mumol/L isoproterenol-evoked increase in lymphocyte cyclic adenosine monophosphate content (as index for lymphocyte beta-adrenoceptor responsiveness) was diminished to a similar extent. Combining all data resulted in a significant positive correlation between myometrial and lymphocyte beta-adrenoceptor densities (r = 0.7303; n = 36; p less than 0.001). It is possible that determination of beta-adrenoceptor function in circulating lymphocytes may be a useful model to monitor myometrial beta-adrenoceptor changes during tocolytic therapy. PMID- 2562944 TI - Pharmacokinetic and pharmacodynamic studies of the H1-receptor antagonist hydroxyzine in the elderly. AB - The pharmacokinetics and pharmacodynamics of the antipruritic H1-receptor antagonist hydroxyzine hydrochloride were studied in nine healthy, fasting subjects (mean age 69.5 +/- 3.7 years) who ingested a single dose of hydroxyzine syrup, 0.7 mg/kg (mean dose 49.0 +/- 6.7 mg). Blood samples were collected hourly for 6 hours, every 2 hours from 6 to 12 hours, at 24 hours, and then every 24 hours for 144 hours. At these times an intradermal injection of 0.01 ml of a 0.1 mg/ml histamine phosphate solution was performed, and wheal and flare areas were computed. The serum elimination t1/2 of hydroxyzine was 29.3 +/- 10.1 hours; the volume of distribution was 22.5 +/- 6.3 L/kg; the clearance rate was 9.6 +/- 3.2 ml/min/kg, and the AUC was 1383.1 +/- 1039.0 ng.hr/ml. The mean serum elimination t1/2 of cetirizine, the active metabolite of hydroxyzine generated in vivo, was 24.8 +/- 7.7 hours, not significantly different from that of the parent compound (p = 0.05). After a single dose of hydroxyzine the mean wheal and flare areas were significantly suppressed from 1 to 144 hours, compared with the mean predose wheal and flare sizes (p less than 0.01). Maximum wheal suppression, compared with all other wheals measured during the study, occurred from 4 to 10 hours, inclusive, and maximum flare suppression occurred from 2 to 72 hours, inclusive (p less than 0.01). Hydroxyzine has a long t1/2 and a large volume of distribution in the elderly. The suppressive effect on the wheal and flare after a single dose of hydroxyzine is also extremely prolonged, suggesting the possibility of enhanced H1-receptor activity in old age. PMID- 2562945 TI - [Diagnostic significance of antineutrophil cytoplasmic antibodies in Wegener's granulomatosis and related disease pictures]. PMID- 2562946 TI - [Therapy of Wegener's granulomatosis and related vasculitides]. PMID- 2562947 TI - AIDS: a syndrome of immune dysregulation, dysfunction, and deficiency. AB - Acquired immune deficiency syndrome (AIDS) is a disease caused by the human immunodeficiency virus (HIV) in which cellular immune functions are severely impaired. Acute infection and subsequent destruction of helper T cells, although occurring readily in cell cultures, do not appear to be the only mechanisms mediating helper T cell loss. Other mechanisms that may account for the loss of helper T cells include: T cell syncytia formation, decreased T cell production, and autoimmune-related destruction of helper T cells. Immune abnormalities seen early in the course of HIV infection include immune hyperactivation and autoimmune phenomena suggestive of immune dysregulation rather than immune deficiency. Many changes in immune function are, in fact, seen in HIV seropositive patients who possess a normal number of helper T cells. Mechanisms (other than the loss of helper T cells) that may contribute to the immune abnormalities seen in these patients include noninfectious effects of HIV and HIV proteins, effects of HIV on non-T cells, autoimmune-related manifestations of HIV infection, and HIV-induced activation of normal immunosuppressive circuits. PMID- 2562948 TI - Ultrasonic patterns observed in hepatic metastases from breast carcinoma: diagnosis and evolution. AB - A retrospective analysis was made of 78 patients presenting breast neoplasm with hepatic metastases confirmed by ultrasound. Clinical hepatomegaly was present in 61%. The serum glutamic-oxaloacetic transaminase (SGOT) was elevated in 72%, the serum glutamic-pyruvic transaminase (SGPT) in 56%, the serum alkaline phosphatase (Aph) in 86%, and the gamma-glutamil transpeptidase (GGT) in 76%. A hypoechogenic multiple nodular pattern (HMN) was observed in 69%, a diffuse hypoechogenic pattern (DH) in 15%, and a mixed multiple nodular pattern (MMN) in 11%. No single nodular pattern was presented in any patient. The univariate analysis showed a better survival rate in patients with a mixed pattern (mean 11 months, range 1-29 months) (p = 0.027). No significant differences were observed regarding the remaining patterns, age, presence or not of hepatomegaly, or altered enzymatic values. PMID- 2562949 TI - Inhibition of antibody secretion by 5-aminosalicylic acid. AB - We have examined the effects of sulfasalazine and its metabolites sulfapyridine and 5-aminosalicylic acid on antibody secretion by normal peripheral blood and intestinal mononuclear cells. Sulfasalazine and 5-aminosalicylic acid both inhibited pokeweed mitogen-stimulated secretion of immunoglobulins (Igs) A, G, and M by peripheral blood mononuclear cells in a dose-dependent manner, whereas sulfapyridine had little effect. Sulfasalazine and 5-aminosalicylic acid also inhibited spontaneous secretion of IgA by intestinal mononuclear cells, but sulfapyridine did not. Sulfasalazine inhibited pokeweed mitogen-stimulated lymphocyte proliferation, while 5-aminosalicylic acid and sulfapyridine exhibited minimal inhibition. Sulfasalazine was toxic for peripheral blood mononuclear cells, whereas 5-aminosalicylic acid and sulfapyridine were not toxic. Thus, the inhibition of antibody secretion by sulfasalazine was due to direct toxicity. On the other hand, 5-aminosalicylic acid, the therapeutically active component of sulfasalazine, was neither toxic nor antiproliferative, and appeared to exert its effects on metabolic pathways directly related to antibody synthesis. The calculated ID50 values of 5-aminosalicylic acid for antibody secretion were 1.35 mM for IgA and 1.05 mM for IgG, concentrations that are achieved in the colons of treated individuals. Indomethacin did not inhibit antibody secretion at pharmacologically relevant concentrations. 5-Aminosalicylic acid mediated inhibition of antibody secretion may play a role in inflammatory bowel disease by stopping antibody-mediated memory events involved in the induction or perpetuation of the disease process. PMID- 2562950 TI - Managing angina in the elderly: an update. AB - Angina pectoris in the elderly is usually due to fixed coronary arteriosclerotic disease. Management includes the use of nitrates and beta- and calcium-blocking drugs. A combination of these drugs may be necessary in patients with severe symptoms. Each of these drugs has potential side effects. These drugs should be started in small dosages and gradually increased, determined by the patient's response. Angina with associated disease states should determine what specific drugs are selected. PMID- 2562951 TI - Mortality from neuroleptic malignant syndrome. AB - The authors assess the mortality from the neuroleptic malignant syndrome (NMS) based on an exhaustive review of 202 published case reports, including a differential assessment of risk factors and protective factors. The results indicate a significant (p less than .05) decrease in mortality since 1984 (11.6% vs. 25% before 1984), which occurs independently of the therapeutic use of dopamine agonists and dantrolene. A significantly (p less than .001) lower rate of mortality from haloperidol-induced NMS (7%) and a high rate of mortality (38.5%) among patients with organic brain syndrome were also found. Myoglobinemia and renal failure are strong predictors of mortality, representing a mortality risk of approximately 50%. The authors discuss the implications of these findings. PMID- 2562952 TI - Effects of verapamil on tardive dyskinesia and psychosis in schizophrenic patients. AB - Nine hospitalized schizophrenic patients with tardive dyskinesia were treated with the calcium-channel antagonist verapamil under single-blind conditions. The tardive dyskinesia and activation scores decreased, and the anxiety/depression scores increased. The changes were small but statistically significant. PMID- 2562953 TI - Triggering of T-lymphocytes via either T3-Ti or T11 surface structures opens a voltage-insensitive plasma membrane calcium-permeable channel: requirement for interleukin-2 gene function. AB - Stimulation of human T-lymphocytes via either the surface T3-Ti antigen-major histocompatibility complex receptor complex or the T11 molecule results in clonal proliferation through a calcium-dependent mechanism. To investigate this signal transduction, plasma membrane calcium-permeable channels were characterized in T lymphocytes by means of whole cell or single channel patch-clamp recordings. Stimulation of T-lymphocytes via either structure results in opening of an identical set of voltage-insensitive plasma membrane Ca2+-permeable channels through the action of a diffusible second messenger. Previous work with excised inside-out patches suggests that inositol 1,4,5-trisphosphate is the activating second messenger of the voltage-insensitive T-cell Ca2+-permeable channel. Since there is a significant increase in phosphoinositide turnover after stimulation via either the T3-Ti or T11 pathway, it is suggested that triggering of either structure opens a common set of channels through this mechanism. Furthermore, currents flowing through Ca2+-permeable channels are apparently autoregulated, as inward conductance is abolished by elevation of Ca2+ concentration in the bathing solution. In particular, the steady-state rise in interleukin-2 (T-cell growth factor) mRNA is dependent on the rise of [Ca2+]i resulting from ion movement across this channel. PMID- 2562954 TI - Subcellular localization of the major pneumococcal autolysin: a peculiar mechanism of secretion in Escherichia coli. AB - The major pneumococcal autolysin (N-acetylmuramoyl-L-alanine amidase) has been localized in the cellular envelope of Streptococcus pneumoniae and Escherichia coli by using immunocytochemical labeling on ultrathin sections and whole-mounted cells. Cell fractionation experiments in E. coli confirmed the peripheral localization of the pneumococcal amidase and suggested that this enzyme is weakly bound to the outer face of the cytoplasmic membrane. This interaction does not depend on the presence of choline but represents an intrinsic property of the amidase. The autolysin, that is synthesized without any N-terminal signal sequence (Garcia, P., Garcia, J. L., Garcia, E., and Lopez, R. (1986) Gene (Amst.) 43, 265-272) was not processed during translocation. A new regulatory mechanism that might be specific for bacterial autolysins is discussed. PMID- 2562955 TI - Cloning and expression of human arylsulfatase A. AB - A full length cDNA for human arylsulfatase A was cloned and sequenced. The predicted amino acid sequence comprises 507 residues. A putative signal peptide of 18 residues is followed by the NH2-terminal sequence of placental arylsulfatase A. One of the arylsulfatase A peptides ends 3 residues ahead of the predicted COOH terminus. This indicates that proteolytic processing of arylsulfatase A is confined to the cleavage of the signal peptide. The predicted sequence contains three potential N-glycosylation sites, two of which are likely to be utilized. The sequence shows no homology to any of the known sequences of lysosomal enzymes but a 35% identity to human steroid sulfatase. Transfection of monkey and baby hamster kidney cells resulted in an up to 200-fold increase of the arylsulfatase A activity. The arylsulfatase A was located in lysosome-like structures and transported to dense lysosomes in a mannose 6-phosphate receptor dependent manner. The arylsulfatase A cDNA hybridizes to 2.0- and 3.9-kilobase species in RNA from human fibroblasts and human liver. RNA species of similar size were detected in metachromatic leukodystrophy fibroblasts of two patients, in which synthesis of arylsulfatase A polypeptides was either detectable or absent. PMID- 2562956 TI - Progesterone interacts with P-glycoprotein in multidrug-resistant cells and in the endometrium of gravid uterus. AB - P-Glycoprotein (P-GP) plays a pivotal role in maintaining the multidrug-resistant (MDR) phenotype. This membrane glycoprotein is overproduced in MDR cells and the endometrium of the mouse gravid uterus (Arceci, R.J., Croop, J.M., Horwitz, S.B., and Housman, D. (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 4350-4354). This latter observation and an interest in endogenous substrates for P-GP led to a study of the interaction of steroids with P-GP found in the endometrium of the mouse gravid uterus and in MDR cells derived from the murine macrophage-like cell J774.2. [3H]Azidopine labeling of P-GP from these two sources was inhibited by various steroids, particularly progesterone. Progesterone also markedly inhibited [3H]vinblastine binding to membrane vesicles prepared from MDR cells, enhanced vinblastine accumulation in MDR cells, and increased the sensitivity of MDR cells to vinblastine. In addition, we have demonstrated that the hydrophobicity of a steroid is important in determining its effect on inhibition of drug binding to P GP. It is concluded that progesterone, a relatively nontoxic endogenous steroid, interacts with P-GP and is capable of reversing drug resistance in MDR cells. PMID- 2562957 TI - Distinct subsets of somatostatin receptors on cultured human lymphocytes. AB - Somatostatin (SOM) is a neuroendocrine tetradecapeptide that suppresses specific functions of differentiated T-cells and antibody-producing cells. The Jurkat line of human leukemic T-cells and U266 IgE-producing human myeloma cells bound [I Tyr11]SOM specifically. The maximal level of specific binding was attained by 1-2 h at 22 degrees C for both types of cells and reversed by 70-85% within 2-3 h after the addition of excess nonradioactive SOM. Computer-assisted Scatchard analysis of the competition curves revealed two classes of binding sites for both cells. An average of 144 and 1295 high affinity receptors per Jurkat and U266 cells had a Kd value of 3 pM and 5 pM, respectively, whereas a large number of low affinity sites had Kd values of 66 nM and 100 nM. The affinity of the analogs somatostatin 28, [I-Tyr11]SOM, and [D-Trp8, D-Cys14]SOM for Jurkat and U266 cell lines, relative to SOM, suggested a degree of specificity similar to receptors on neuroendocrine cells. PMID- 2562958 TI - Hypoxia-induced dysfunctions and injury of astrocytes in primary cell cultures. AB - The effects of severe hypoxia were studied in a primary culture of astrocytes prepared from newborn rat cerebral cortex. Hypoxia was created by placing cultures in an airtight chamber that was flushed with 95% N2/5% CO2 for 15 min before being sealed. The hypoxic environment was maintained constant for up to 24 h. During the first 12 h of hypoxia, astrocytes showed no morphological changes by phase-contrast microscopy. After 18 h of hypoxia, some astrocytes in culture became swollen and started to detach from the culture dish. All cells in the culture were destroyed after 24 h of hypoxia. The lactate dehydrogenase level in the culture medium increased more than tenfold between 12 and 24 h of hypoxia. Glutamate uptake was inhibited 80% by similar hypoxic conditions. The cell volume of astrocytes, as measured by 3-O-methyl-[14C]-D-glucose uptake, was increased. These observations suggested cell membrane dysfunction. The malondialdehyde level of hypoxic cultures increased two-fold after 24 h of hypoxia. Verapamil (0.5 mM), furosemide (1 mM), indomethacin (1 mM), MgCl2 (10 mM), and mannitol (10 mM) reduced but never completely abolished the release of lactate dehydrogenase from hypoxic astrocytes. These data suggest multifactorial causes for severe injury in hypoxic astrocytes. PMID- 2562959 TI - Responsiveness of superficial hand veins to phenylephrine in essential hypertension. Alpha adrenergic blockade during prazosin therapy. AB - Patients with essential hypertension show an increase in vascular resistance. It is unclear whether this is caused by structural changes in the arterial wall or by hyperresponsiveness of vascular smooth muscle to endogenous alpha adrenergic agonists. Using the dorsal hand vein compliance technique we compared the changes in diameter of superficial veins in response to phenylephrine, an alpha 1 adrenergic receptor agonist, and to nitroglycerin, a venorelaxant, in patients with essential hypertension and in normotensive subjects. The dose of phenylephrine that produced 50% of maximal venoconstriction (ED50) in the hypertensive subjects was 257 ng/min (geometric mean; log mean +/- SD was 2.41 +/ 0.54). In the control subjects the ED50 was 269 ng/min (geometric mean; log mean was 2.43 +/- 0.43). Maximal response (Emax) for phenylephrine was 84 +/- 13% in the hypertensive subjects and 90 +/- 6% in the control subjects. Differences in the group means of the ED50 (P = 0.92) or the Emax (P = 0.27) were not significant. There were no significant differences in the ED50 (P = 0.54) or the Emax (P = 0.08) for nitroglycerin between the two groups. These results show no evidence for a generalized change in alpha adrenergic responsiveness in hypertension and support the concept that increased blood pressure responses to alpha adrenergic stimulation in hypertensives are due to structural and geometric changes in the arterial wall rather than to an increased responsiveness of postsynaptic alpha adrenergic receptors. The phenylephrine studies were repeated in seven hypertensive patients during treatment with prazosin, an alpha 1 adrenergic antagonist. The mean dose ratio of the shift in phenylephrine ED50 (ED50 during prazosin therapy/ED50 before prazosin therapy) was 6.1. This indicates that small doses of prazosin (1-2 mg) cause significant in vivo shifts in the dose-response relationship of alpha adrenergic agonists. The dorsal hand vein compliance technique is useful in detecting systemic effects of alpha adrenergic antagonists. PMID- 2562960 TI - Genotypes for aldehyde dehydrogenase deficiency and alcohol sensitivity. The inactive ALDH2(2) allele is dominant. AB - Many Orientals lack the mitochondrial aldehyde dehydrogenase (ALDH2) activity responsible for the oxidation of acetaldehyde produced during ethanol metabolism. These individuals suffer the alcohol-flush reaction when they drink alcoholic beverages. The alcohol-flush reaction is the result of excessive acetaldehyde accumulation, and the unpleasant symptoms tend to reduce alcohol consumption. The subunit of this homotetrameric enzyme was sequenced and the abnormality in the inactive enzyme shown to be a substitution of lysine for glutamate at position 487. We have used the polymerase chain reaction to determine the genotypes of 24 livers from Japanese individuals. Correlating genotype with phenotype leads to the conclusion that the allele (ALDH2(2)) encoding the abnormal subunit is dominant. PMID- 2562961 TI - MR, CT, and ultrasound findings of metastatic vipoma in pancreas. AB - Magnetic resonance, CT, and ultrasound findings are described for a vipoma of the pancreas with hepatic metastases in a patient with Verner-Morrison syndrome and multiple endocrine neoplasia I. PMID- 2562962 TI - Musical hallucinations. The sounds of silence? AB - Hallucinations may occur in any sensory modality. Auditory hallucinations, usually ascribed to psychiatric illness, take various forms including the perception of voices, cries, noises, or rarely, music. Formed musical hallucinations, (ie, the perception of either vocal or instrumental melodies), reported in the English literature to date have typically been associated with marked hearing loss, advanced age (average 67.8 years), female sex (71%), lack of response to treatment, and general lack of associated psychopathology. We have collected data on seven additional patients with musical hallucinations. The average age of these patients was 72.9 years; all were women. Six had significant hearing problems. All reported onset of musical hallucinations after the age of 60. Interestingly, all seven had major psychiatric illnesses. Four had major depression, two had late-onset schizophrenia, and one had multi-infarct dementia. Of the five who had CT scans, one was normal and the rest demonstrated varying degrees of brain pathology. Neuroleptics were used with varying results in three cases; antidepressants were used in two depressed patients and were temporally related to the onset of musical hallucinations in one patient. Electroconvulsive therapy (ECT) was very effective in treating depression and musical hallucinations in the three patients for whom it was used, usually providing relief from hallucinations after only two treatments. Our collection of cases demonstrates that musical hallucinations can occur in association with psychiatric illness, and perhaps unlike the hallucinations associated with isolated hearing loss, may respond to conventional treatments for the underlying psychiatric disorder. Hearing loss is neither a necessary nor sufficient condition for the occurrence of musical hallucinations. PMID- 2562963 TI - Two monoclonal antibodies identify thymic-repopulating cells in mouse bone marrow. AB - The progenitor cells in the bone marrow that home to and repopulate the thymus have been incompletely characterized. In particular, it is not clear whether thymocytes differentiate directly from pluripotent hemopoietic stem cells that seed to the thymus, or whether T lymphoid-committed stem cells (prothymocytes) arise in the bone marrow before the thymic migration. In order to resolve this question, we have used mAb specific for cell-surface Ag to identify the bone marrow cells which can seed to and repopulate the thymus of irradiated mice. We report here that the majority of thymic-repopulating cells in mouse bone marrow express low levels of the Thy-1 Ag (Thy-1lo) plus high levels of a newly described Ag termed stem cell Ag (Sca-1). Two distinct populations of thymic repopulating Thy-1loSca-1+ cells in mouse bone marrow can be discriminated based on expression of any of a number of hemolymphoid lineage-specific (Lin) markers. Thus, Thy-1loLin-Sca-1+ and Thy-1loLin+Sca-1+ fractions of bone marrow contain thymic-repopulating cells. A second Ag, stem cell Ag-2 (Sca-2), is expressed by Thy-1loLin+Sca-1+ cells but not by Thy-1loLin-Sca-1+ cells. The Thy-1loLin-Sca-1+ fraction expresses intermediate levels of the phagocyte glycoprotein-1 Ag, and comprises 30% of the Thy-1loLin- bone marrow cells, which have previously been shown to be highly enriched in pluripotent hemopoietic stem cells. By facilitating the isolation of highly purified subpopulations of bone marrow cells that can repopulate the thymus, Sca-1 and Sca-2 should provide an experimental tool for describing the developmental potential of such bone marrow subsets. PMID- 2562964 TI - Regulation of idiotope expression. IV. Genetic linkage of two D region-dependent T15 idiotopes to the IgH allotype. AB - The idiotopic (Id) repertoire of antibody response to phosphocholine was studied in mouse strains with different IgH allotypes. The T15 idiotype-bearing (T15+) serum antibody and antibody plaque-forming cells (PFC) were characterized with four monoclonal anti-Id that recognize distinct Id determinants on T15+ antibody encoded by VH-1 (of the S107 gene family), DH FL16.1, JH-1 and Vk22 germ-line genes. We have previously shown that expression of the Id designated AB1-2 and B36-82 depends on the third hypervariable loop (D region), whereas the other Id, MaId5-4 and B24-44, are influenced by VH structures outside of the D region. All four Id were expressed in the PC-response of all mouse strains tested, except the Ighj strains (C3H/HeJ, CBA/H-T6, PL/j), where the D region-dependent Id, AB1-2 and B36-82, were absent. The other Id, however, were normally expressed on individual PFC as well as the serum antibody of the Ighj strains. Expression of AB1-2 and B36-82 on 50% of PFC occurred in (BALB/c-Igha x C3H/HeJ-Ighj)F1 mice. The absence of Id correlated with a unique RFLP of the S107 gene family in Ighj strains. Finally, Id expression segregated with the appropriate RFLP pattern in individual (BALB/c x C3H/HeJ)F2 mice. These data demonstrate a selective genetic linkage of discrete T15 Id determinants, AB1-2 and B36-82 with the Igh allotype. By comparing these results with the available Ig sequences, we suggest that the Ighj allotype may be associated with an allelic form of the DH-FL16.1 segment which with VH-1, JH-1, and the Vk 22 code for the phosphocholine-specific antibody in the mouse. PMID- 2562965 TI - Improved cell depletion in a panning technique using covalent binding of immunoglobulins to surface modified polystyrene dishes. AB - A method for the chemical modification of plastic surfaces permits covalent binding of proteins and we have used this method in the development of an efficient panning technique. Thus, human peripheral T lymphocytes coated with mouse monoclonal antibodies directed against the CD4 marker may be selectively and reproducibly removed from a lymphocyte population by a short incubation in modified plastic dishes coated with rabbit anti-mouse IgG antibody. Due to the higher protein binding capacity of the dishes the use of the IgG fraction of the coating antibody was sufficient for optimal and reproducible results. In contrast, control dishes with passively adsorbed antibody required an affinity purified fraction and even then were less efficient. PMID- 2562966 TI - Dose dependent latency of fatal gastrointestinal and bone marrow syndromes. AB - Two acutely-responding normal tissues, bone marrow and gastrointestinal epithelium, have been shown to exhibit a biphasic dose-latency response to radiation with both a dose-dependent and dose-independent component for expression of radiation injury. PMID- 2562967 TI - Trypsinization and the radiosensitivity of mitotic and log phase Chinese hamster V79 cells exposed to 250 kVp X-rays. AB - We studied the influence of trypsin-induced morphological changes on the radiosensitivity of cells plated at either low (4-600/cm2) or high (2 x 10(4)/cm2) density and grown overnight before treatments. Trypsin treatment induced contraction and rounding of spread cells. The radiosensitivity of cells trypsinized and plated either: (1) immediately before [(a) D0 = 1.7 Gy for cells at low-, and (b) 1.5 Gy at high-density] or (2) immediately after (D0 = 1.6 Gy, high-density cells) irradiation was higher than that of (3) cells at high density, irradiated and delayed plated [cells remained spread until the completion of potentially lethal damage repair (PLDR) and trypsinization, D0 = 2.2 Gy], and (4) cells at low density which were neither delayed plated nor trypsinized (i.e. remained spread) after irradiation (Do = 2.4 Gy). These data show that PLDR is reduced in trypsin-treated cells of both high (compare 1b and 2 with 3) and low (compare 1a with 4) density cultures; the latter comparison provides a direct measure of the trypsin effect. Since the comparison between conditions 1 and 2 vs. 3 and 4 is of round vs. spread cells, PLDR appears to be influenced by the cell's morphological state. Kinetic studies showed that when cells were incubated in growth medium to recover from trypsin-induced effects before irradiation, the radiation sensitivity of spread cells (plated in situ), but not of those remaining rounded (in suspension until plating and irradiation), decreased and became equal to that of delayed plated high-density cells. Neither irradiated cells treated with hypertonic saline, nor mitotic cells, showed the trypsin effect. From these results we suggest that: (1) trypsin-induced cell contraction affects the ability of cells to repair radiation damage, (2) spread cells are better able to repair PLD than rounded cells, (3) immediate plating survival of cells in high-density cultures may not represent their intrinsic radiosensitivity and (4) cell-to-cell contact is not necessary for log phase cells to repair PLD. PMID- 2562968 TI - Recovery from radiation damage in human squamous carcinoma of the cervix. AB - The radiobiology of human carcinoma of the cervix has been investigated using five recently established cell lines. By means of an in vitro clonogenic assay, cell survival has been determined at dose-rates of 150, 3.2 and 1.6 cGy/min, and, in addition, split-dose experiments have been performed. Using approximately isoeffective doses (1 per cent cell survival), the amount of recovery observed using dose-rate and split-dose experiments has been correlated with the surviving fraction at 2 Gy (SF2 value) and the parameter alpha from the acute cell survival curves for the five lines. Two lines were radiosensitive, possessing significantly lower SF2 values (p = 0.03), and, at these isoeffect doses, showed significantly less sparing at low dose-rate (p = 0.012) and less split-dose recovery (p = 0.0096) than the three remaining cell lines. In general, the results indicate a good correlation between split-dose recovery-ratio, dose sparing achieved at low dose-rate compared to high, and SF2 value, and an inverse correlation with the alpha value derived from fitting dose rate dependence data to the incomplete-repair model of Thames. These observed differences in radiosensitivity within one human tumour type indicate the possible value of predictive testing of intrinsic radiosensitivity in the radiotherapy of carcinoma of the cervix. PMID- 2562969 TI - Radiological protection and the lymphatic system: the induction of leukaemia consequent upon the internal irradiation of the tracheobronchial lymph nodes and the gastrointestinal tract wall. AB - The excess of leukaemia among young people living in the vicinity of the nuclear fuel reprocessing plant at Sellafield has focused attention on the possibility that irradiation of the lymphatic system from particulate alpha-emitting nuclides might be responsible. We discuss below two possible routes of such exposure; namely the inhalation and ingestion of particulates. We conclude that, in spite of the real possibility of substantial doses to tissues associated with the lymphatic system, there is little reason to expect that lymphatic leukaemia will be the dominant outcome of the exposure. However, the arguments presented are not, and cannot be, wholly conclusive. PMID- 2562970 TI - Efficacy and toxicity of N,N',N",N"'-tetra(2,3,4-trihydroxybenzoyl)-spermine for decorporation of 239Pu from mice. AB - Male SAS/4 mice were injected i.v. with 6.6 kBq 239Pu-citrate. After 1 or 24 h a single i.p. injection of 15 or 30 mumol kg-1 or repeated (three or four) daily injections of 30 mumol kg-1 of tetra-THB-spermine were given, and at 4 or 7 days Pu retention was measured in liver, kidneys and femur. Besides tetra-THB spermine, equimolar doses of tetra-DHB-spermine were injected for comparison, or equimolar doses of diethylene triamine-pentaacetic acid (DTPA) as a reference compound. Histological changes in kidneys and liver were examined after i.p. injections of 30 mumol kg-1 or at 2-13 times higher doses of tetra-THB-spermine. The results show that: (1) Introduction of an additional hydroxy group into the aromatic moieties of tetra-DHB-spermine results in increased hydrophilicity, lower toxicity and a lower renal retention of Pu. (2) Tetra-THB-spermine and tetra-DHB-spermine are similarly effective in removing plutonium from liver and bone. Their efficacies in removing Pu from bone are approximately similar to those of DTPA but for whole-body removal they are generally inferior. (3) Multiple (30 mumol kg-1) of tetra-THB-spermine were no more effective than a single injection at mobilizing Pu from the liver. (4) Four injections of tetra THB-spermine induced cloudy swelling and fatty degeneration in epithelial cells of the proximal convoluted tubules. At levels of 400 mumol kg-1 tetra-THB spermine produced severe degenerative glomerular lesions, foci of liver necrosis and thromboses of the portal vein branch. PMID- 2562971 TI - Radiation damage to glucose concentrating capacity and cell survival in kidney tubule cells: effects of fractionation. AB - The glucose concentrating capacity of cultured LLC-PK1 kidney epithelial cells has been measured after single and fractionated doses of X-rays. Steady-state glucose concentrating capacity (ratio of glucose concentration inside to outside cell) can be measured using radiolabelled analogues of glucose which are actively transported but not metabolized. These cells can be stimulated to increase their glucose concentrating capacity (up-regulation) by a reduction in the glucose concentration of the growth medium. However, after X-ray irradiation the cells have a reduced capacity to respond to up-regulation. This effect can be measured 7 days after irradiation and before radiation-induced cell killing affects the cell population. The previously reported radiosensitivity of this function to single doses of X-rays (in the range 1-16 Gy) was confirmed. Surprisingly, no significant sparing of this effect could be measured by fractionation of the X ray dose into two or four fractions. However, the cells showed a significant fractionation effect if clonogenic survival was measured using the standard cell survival assay. These early effects have different fractionation response from the later phases of tissue damage, measured months to years after irradiation, which do show sparing due to fractionation and are thought to be mainly due to changes in cell survival. The lack of sparing by fractionation to the functional damage may suggest a different target from that which determines cell survival. These results support the hypothesis that radiation damages cellular functions, separately from cell replication. PMID- 2562973 TI - Lethal mutations in cultured mammalian cells. PMID- 2562972 TI - Are solitons responsible for energy transfer in oriented DNA? PMID- 2562974 TI - Direct estimation of latent time for radiation injury in late-responding normal tissues: gut, lung, and spinal cord. AB - Mixture models are proposed for simultaneous analysis of the latency and fractionation characteristics of radiation injury in late-responding normal tissues. The method is an extension of the direct analysis for quantal response data. Conceptually, the application of the mixture model is based on the biological observation that over a wide range of doses a proportion of the irradiated subjects will never express damage. Mixture models allow the time of occurrence to be utilized in the analysis. Furthermore, this type of model takes time-censored observations into account in a natural way and provides an adequate framework for modelling and analysis of effect-dependent latency. Mixture models with complete and incomplete repair are applied to dose-incidence data for four late endpoints in rodents: death from radiation-induced pneumonitis, leg paralysis after spinal-cord irradiation, and radiation-induced rectal stenosis and anal discharge. Radiation-induced pneumonitis had an effect-dependent latency. The modelling of this phenomenon correlates well with the results of histologic studies. Interestingly, the ratio of hazard rates was not constant for this endpoint. The dominating feature in the latency of radiation injury to the spinal cord was a strong dependency on dose per fraction. After correction for this effect a tendency towards a longer latent time for lower effect levels was observed. For the rectal complications, there was no difference between latency with radiation only vs. radiation combined with cis-platin. PMID- 2562975 TI - Pulse radiolysis of catalase in solution. I. Reactions of O2- with catalase and its compound I. AB - The time-course of absorption changes of oxygen-saturated solutions of bovine liver catalase after pulse radiolysis have been studied. The rate constant of formation of Compound I due to the reaction of catalase with hydrogen peroxide has been estimated to be 2.0 x 10(7) dm3mol-1s-1. Radiation generated superoxide radicals reduce Compound I to Compound II with a rate constant of 5.0 x 10(6) dm3mol-1s-1. The formation of Compound III in the direct reaction of O2- with catalase has also been observed. PMID- 2562976 TI - OH-induced free radicals in purine nucleoside monophosphates: e.s.r. and spin trapping. AB - Free radicals produced by the reactions of OH radicals with six purine nucleoside monophosphates (3'-AMP, 5'-AMP, 5'-dAMP, 3'-GMP, 5'-GMP and 5'-dGMP) were investigated by a method combining e.s.r. spin-trapping and high-performance liquid chromatography (HPLC). The N2O-saturated aqueous solutions of purine nucleoside monophosphates, containing 2-methyl-2-nitrosopropane as a spin-trap, were X-irradiated and the resulting spin-adducts were separated by reverse-phase HPLC in the ion suppression mode. The separated spin-adducts were characterized by e.s.r. spectrometry and UV spectrophotometry. Consequently, the radicals due to H-abstraction at the C4' position of the sugar moiety were identified arising from 5'-dAMP and 5'-dGMP. In all cases, e.s.r. spectra consisting of a secondary doublet were observed and assigned to the radical due to H-abstraction at the C5' position of the sugar moiety. PMID- 2562977 TI - Photokinetic and ultrastructural studies on porphyrin photosensitization of HeLa cells. AB - Liposome-bound haematoporphyrin or haematoporphyrin dimethylester, as well as haematoporphyrin dissolved in phosphate-buffered saline, were added to HeLa cell monolayers at a dose of 1 microgram of porphyrin per 10(5) cells. After 2 min or 20 min incubation liposome-bound porphyrins were accumulated by cells in an about two-fold larger amount than the water-dissolved haematoporphyrin. This caused a more efficient photosensitization of HeLa cells by liposome-delivered porphyrins upon illumination with 366 nm light. Ultrastructural studies of HeLa cells, which had been incubated in a physiological medium for 24h after the end of irradiation, showed that liposomal porphyrins induce an early and extensive endocytoplasmic damage, leading to swelling of the mitochondria and vesiculation; changes of the permeability of the cytoplasmic membrane are also evident, especially in the case of haematoporphyrin dimethylester. On the other hand, water-dissolved haematoporphyrin predominantly photosensitizes damage of the plasma membrane. The different pattern of cell photodamage probably reflects a different subcellular distribution of the photosensitizing drugs. PMID- 2562978 TI - Failure to detect immune deficiency in rats after prenatal or early postnatal irradiation. AB - We have looked for medium-term sequelae in the immune system of rats that had been X-irradiated (0-2 Gy whole-body irradiation) during prenatal or early postnatal life. At an age of 8 weeks the histology of the spleen was normal, and so was the distribution of B and T lymphocytes. The serum immunoglobulin levels were not significantly altered, even when the different isotypes were considered. At an age of 10 weeks, the rats were immunized with a T-dependent or a T independent dinitrophenylated-carrier antigen. Normal levels of specific antibodies were generated in all groups of animals injected with the T independent antigen. The T-dependent response, in contrast, was higher in animals irradiated between day 6 and day 20 of gestation (but not in rats irradiated early after birth). This increase, however, was significant only for the IgM and IgG1 responses of some irradiated groups. Thus no medium-term immunodeficiency could be documented with the methods used. The alteration in a T-dependent response, however, points to a radiosensitive T regulatory mechanism. PMID- 2562979 TI - Kinetics of chromatid aberrations in G2 ataxia-telangiectasia cells exposed to X rays and ara A. AB - The cytogenetic effects of X-rays alone or in combination with 9-beta-D arabinofuranosyladenine (ara A) were studied in an immortalized fibroblastic line of ataxia-telangiectasia (A-T) cells. The average length of G2 in this line was determined by autoradiographic labelling (labelled mitoses) to be approximately 5 h. Samples of A-T cells treated with or without ara A, 4 h prior to fixation were irradiated at 1/2-hourly intervals, from 1.5 h to 3.5 h before fixation and then examined for the presence of metaphase chromatid aberrations. It is postulated that the kinetics of disappearance (rejoining) of chromatid deletions with postirradiation incubation time reflects the underlying repair of dsb. This rejoining was found to be inhibited by ara A. Thus the frequency of deletions in the presence of ara A should represent the frequency of deletions in the absence of dsb repair. The rejoining kinetics for deletions in A-T was similar to that found in a previous study of normal human fibroblasts (Mozdarani and Bryant 1987). The number of deletions in X-irradiated A-T cells at 1.5 h before fixation was found to be higher by a factor of approximately 2 than that found previously in normals, indicating that in A-T a higher rate of conversion of dsb into chromatid deletions occurs. The frequency of exchanges induced in G2 A-T cells was similarly enhanced but, unlike the situation in normal cells, ara A was found to cause only a slight increase in this frequency. PMID- 2562980 TI - Increased chromosome aberration levels in cells from mouse fetuses after zygote X irradiation. AB - Cell cultures derived from skin biopsies of mouse fetuses X-irradiated at the zygote stage (2 Gy) were studied for the presence of chromosomal aberrations and micronuclei. In comparison with cells from control, unirradiated fetuses significantly higher spontaneous frequencies of aberrations per 100 metaphases and aberrant metaphases were found in cells which were obtained from normal fetuses and fetuses with gastoschisis after irradiation of the zygote. The proportion of aberrant metaphases in cells from fetuses with gastroschisis but not the frequency of aberrations per 100 cells was significantly higher than in cells from normal fetuses. Spontaneous aberrations were mainly chromatid and chromosome fragments. A significantly higher proportion of cells with micronuclei was observed in cells from fetuses with gastroschisis than in cells from control or normal fetuses after irradiation. The induction of chromosome instability in fetal fibroblasts after zygote irradiation has not previously been reported. PMID- 2562981 TI - RBE-LET relationship for the survival of V79 cells irradiated with low energy protons. AB - The survival of V79 Chinese hamster cells irradiated with proton beams with energies of 0.73, 0.84, 1.16, 1.70 and 3.36 MeV, corresponding to LET values, evaluated at the cell midplane, of 34.5, 30.4, 23.9, 17.8 and 10.6 keV/micron respectively, have been studied in the dose range 0.5-6.0 Gy. As a reference, the survival curve obtained with 200 kV X-rays was used. The initial shoulder, typical of survival curves obtained with sparsely ionizing radiation, decreases as the LET increases and completely disappears at 23.9 keV/micron. This value corresponds to the maximum of the RBE, expressed as the initial slope ratio. In the energy range we have considered, the RBEs for protons are higher than those reported for other ions of comparable LET and the RBE-LET relationship results shifted to lower LET values. Our data seem to indicate that the RBE-LET curve depends on the type of radiation and this could imply that LET is not a good reference for the dose-effectiveness relationship. PMID- 2562982 TI - FBL-reactive CD8+ cytotoxic and CD4+ helper T lymphocytes recognize distinct Friend murine leukemia virus-encoded antigens. AB - Immunization of C57BL/6 (B6) mice with FBL, a Friend murine leukemia virus (F MuLV), induces both tumor-specific cytolytic CD8+ (CTL) and lymphokine-producing CD4+ Th that are effective in adoptive therapy of B6 mice bearing disseminated FBL leukemia. The current study evaluated the F-MuLV antigenic determinants expressed on FBL that are recognized by FBL-reactive CD8+ and CD4+ T cells. To identify the specificity of the FBL-reactive CD8+ CTL, Fisher rat embryo fibroblast (FRE) cells transfected with plasmids encoding F-MuLV gag or envelope (env) gene products plus the class I-restricting element Db were utilized. FBL reactive CTL recognized FRE target cells transfected with the F-MuLV gag-encoded gene products, but failed to recognize targets expressing F-MuLV env. Attempts to generate env-specific CD8+ CTL by immunization with a recombinant vaccinia virus containing an inserted F-MuLV env gene were unsuccessful, despite the generation of a cytolytic response to vaccinia epitopes, implying that B6 mice fail to generate CD8+ CTL to env determinants. By contrast, CD4+ Th clones recognized FRE target cells transfected with env and not gag genes, and immunization with the recombinant vaccinia virus induced an env-specific CD4+ T cell response. These data show that in a Friend retrovirus-induced tumor model in which tumor rejection can be mediated by either CTL or Th, antigens derived from discrete retroviral proteins are predominantly responsible for activation of each T cell subset. PMID- 2562983 TI - Chromosomal organization of the human major histocompatibility complex class I gene family. AB - 17 HLA class I genes have been isolated from the genome of B-lymphoblastoid cell line 721. Sequence analysis and transfection studies indicate that three genes, in addition to those encoding the HLA-A, -B, and -C antigens can direct the synthesis of a class I alpha protein (4, 5, 21). Using gene-specific DNA probes to analyze the presence of restriction fragment-length polymorphisms within a large pedigree and in panel of HLA deletion mutant cell lines, we show here that two of these genes, designated HLA-G and HLA-F, are located on the short arm of chromosome 6 telomeric to the HLA-A locus. The third expressed non-A, -B, and -C class I gene, HLA-E, is located between HLA-A and HLA-C (4). In addition, the remaining 11 class I pseudogenes and gene fragments are localized relative to established markers on chromosome 6p. PMID- 2562984 TI - Mixed chimerism and permanent specific transplantation tolerance induced by a nonlethal preparative regimen. AB - The use of allogeneic bone marrow transplantation as a means of inducing donor specific tolerance across MHC barriers could provide an immunologically specific conditioning regimen for organ transplantation. However, a major limitation to this approach is the toxicity of whole body irradiation as currently used to abrogate host resistance and permit marrow engraftment. The present study describes methodology for abrogating host resistance and permitting marrow engraftment without lethal irradiation. Our preparative protocol involves administration of anti-CD4 and anti-CD8 mAbs in vivo, 300-rad WBI, 700-rad thymic irradiation, and unmanipulated fully MHC-disparate bone marrow. B10 mice prepared by this regimen developed stable mixed lymphohematopoetic chimerism without any clinical evidence of graft-vs.-host disease. Engraftment was accompanied by induction of specific tolerance to donor skin grafts (B10.D2), while third-party skin grafts (B10.BR) were promptly rejected. Mice treated with the complete regimen without bone marrow transplantation appeared healthy and enjoyed long term survival. This study therefore demonstrates that stable mixed chimerism with donor-specific tolerance can be induced across an MHC barrier after a nonlethal preparative regimen, without clinical GVHD and without the risk of aplasia. PMID- 2562986 TI - Release of glutamate, aspartate, and gamma-aminobutyric acid from isolated nerve terminals. PMID- 2562985 TI - Mouse thymic virus (MTLV). A mammalian herpesvirus cytolytic for CD4+ (L3T4+) T lymphocytes. AB - Mouse thymic virus (MTLV; ICTV designation murid herpesvirus 3) infects developing T lymphocytes of neonatal mice, causing thymic necrosis and acute immunosuppression. Infected animals shed virus indefinitely. In the present report, two-color flow cytometric analysis of T lymphocyte subpopulations defined by the markers CD4 (L3T4) and CD8 (Lyt-2) was used to determine whether MTLV was lytic for a specific thymocyte population. At peak necrosis (8-11 d after infection), numbers of CD4+8+ cells in the thymus were reduced by 80% or more as compared with controls, and CD4+8- cells were reduced by greater than 98%. The major survivors were CD4-8+ and CD4-8- lymphocytes. These data indicate that the CD4 bearing lymphocyte is a primary target for cytolysis during MTLV infection. Possible parallels between MTLV and a newly described lymphotropic human herpesvirus, human herpesvirus 6 (HHV-6/HBLV), are also suggested. PMID- 2562987 TI - Depolarization increases chloride-dependent glutamate sequestration in synaptic membranes of rat cerebral cortex. AB - To assess the functions of Cl- -dependent glutamate "binding" (Cl- -dependent glutamate uptake) in synaptic membranes, possible effects of depolarization on the uptake were examined. When rat cerebral cortical slices were preincubated with depolarizing agents such as veratrine (7 micrograms/ml), 10 microM aconitine, 56 mM K+, and 50 microM monensin, [3H]glutamate uptake by the crude synaptic membranes, which were subsequently prepared from the pretreated slices, was increased by 60-85%. Stimulation of the glutamate uptake by predepolarization was dependent on Na+ but not on Ca2+. The bindings of gamma-[3H]aminobutyric acid and 5-[3H]hydroxytryptamine were not significantly affected by the predepolarization. Veratrine pretreatment increased the maximal density of the glutamate uptake sites without affecting the affinity for glutamate. Several characteristics of the uptake sites increased by the veratrine pretreatment coincided with those of Cl- -dependent glutamate uptake sites. Na+-dependent glutamate binding (Na+-dependent glutamate uptake) to the membranes was not affected by pretreatment with veratrine. The content of endogenous glutamate and the noninulin space in the membrane fractions were not changed by the predepolarization. The increase in the glutamate uptake induced by pretreatment with high K+ was reversible: it returned to the control level after a second incubation of the slices in control medium. These results suggest that the Cl- dependent glutamate sequestration system in synaptic membranes is regulated by the membrane potential. PMID- 2562988 TI - Mechanism of agonist-induced down-regulation and subsequent recovery of muscarinic acetylcholine receptors in a clonal neuroblastoma x glioma hybrid cell line. AB - The mechanisms of carbachol-induced muscarinic acetylcholine receptor (mAChR) down-regulation, and recovery following carbachol withdrawal, were studied in the neuroblastoma x glioma hybrid NG108-15 cell line by specific ligand binding assays. N-[3H]Methylscopolamine ([3H]NMS) and [3H]quinuclidinyl benzilate ([3H]QNB) were used as the ligands for the cell surface and total cellular mAChRs, respectively. Exposure of cells to 1 mM carbachol for 16 h decreased the specific binding of [3H]NMS and [3H]QNB by approximately 80%. Bacitracin (1-4 mg/ml) and methylamine (1-15 mM), inhibitors of transglutaminase and of endocytosis, prevented agonist-induced loss of surface mAChRs. Pretreatment of cells with the antimicrotubular agents nocodazole (0.1-10 microM) and colchicine (1-10 microM) prevented carbachol-induced loss of [3H]QNB binding, but not that of [3H]NMS binding. These results indicate that agonist-induced mAChR down regulation occurs by endocytosis, followed by microtubular transport of receptors to their intracellular degradation sites. When carbachol was withdrawn from the culture medium following treatment of cells for 16 h, receptors recovered and were incorporated to the surface membrane. This recovery process was antagonized by monovalent ionophores monensin (0.1 microM) and nigericin (40 nM), which interfere with Golgi complex function. Receptor recovery was also prevented by the antimicrotubular agent nocodazole. Thus, recovery of receptors appears to be mediated via Golgi complex and microtubular transport to the surface membrane. PMID- 2562989 TI - Translocation and activation of protein kinase C in striatal neurons in primary culture: relationship to phorbol dibutyrate actions on the inositol phosphate generating system and neurotransmitter release. AB - The actions of the tumor-promoting phorbol ester phorbol dibutyrate were examined, under identical physiological conditions, on three distinct cellular processes in striatal neurons: the distribution of protein kinase C, the carbachol-stimulated generation of [3H]inositol monophosphate, and the KCl-evoked release of gamma-[3H]aminobutyric acid ([3H]GABA). Phorbol dibutyrate induced a rapid (complete in 5 min), dose-dependent, entirely reversible (t0.5 = 15 min) translocation of protein kinase C from cytosol to membrane. On longer exposure to phorbol dibutyrate, membrane-associated protein kinase C returned toward the control level, and total cellular enzyme activity declined markedly. Phorbol dibutyrate also induced the dose-dependent attenuation of carbachol-stimulated [3H]inositol monophosphate production and potentiation of KCl-evoked release of [3H]GABA. The translocation of protein kinase C and the potentiation of KCl evoked [3H]GABA release were both rapidly reversed following washout of phorbol dibutyrate. In addition, for both processes, the effect of a 1-h exposure to phorbol dibutyrate was markedly less than that observed following a 5-min exposure to the agent. In direct contrast, inhibition of carbachol-stimulated [3H]inositol monophosphate production was not rapidly reversed following washout of phorbol dibutyrate and was actually more pronounced following a 1-h exposure, compared with a 5-min exposure. These findings indicate that some, but not all, of the actions of phorbol dibutyrate are closely associated with the translocation of protein kinase C in striatal neurons in primary culture. PMID- 2562990 TI - Acute choline supplementation in vivo enhances acetylcholine synthesis in vitro when neurotransmitter release is increased by potassium. AB - The main objective of these studies was to determine whether the acute administration of choline to rats provides supplemental precursor that can be used to support acetylcholine synthesis when the demand for choline is increased by increasing neurotransmitter release. For these experiments, hippocampal and striatal slices were prepared form rats that had received saline or an acute injection of choline. Slices were incubated in a choline-free buffer containing 4.74-35 mM KCl, and acetylcholine synthesis and release and choline production were measured. The initial tissue contents of acetylcholine and choline did not differ between experimental groups for either brain region. When hippocampal slices from the controls were incubated for 10 min with depolarizing concentrations of KCl, acetylcholine release increased and the tissue content decreased in a concentration-dependent fashion; no net synthesis of acetylcholine occurred. In contrast, hippocampal slices from the choline-injected animals maintained their tissue content in the presence of high concentrations of KCl, despite an increase in acetylcholine release that was similar in magnitude to that of the controls; positive net synthesis of acetylcholine resulted. Although the molar concentration of choline achieved in the incubation media at the end of the 10-min period did not differ between groups, the mobilization of free choline from bound stores was significantly greater in hippocampal slices from the choline-injected group than the controls. In addition, the synthesis of acetylcholine by hippocampal slices from the choline-injected group was prevented by the presence of hemicholinium-3 (1 microM) in the media.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2562991 TI - Examining nursing personnel costs: controlled versus noncontrolled oral analgesic agents. AB - To date, few evaluations have focused on nursing personnel costs generated by practices of other clinicians. The authors used traditional industrial engineering and cost accounting techniques to determine nursing personnel costs generated by the use of controlled versus noncontrolled oral analgesic agents. The findings indicate that additional nursing labor costs are generated for drug administration and inventory activities for controlled agents when compared with noncontrolled agents. On an annual basis, the additional costs for drug administration generate approximately +34,000 in hospital expenses for the three nursing units included in this evaluation. This type of interdisciplinary investigation may be valuable as nursing departments face increasing pressures to decrease costs by reducing overtime and eliminating positions. PMID- 2562993 TI - A case of Takayasu's disease with ruptured thoracoabdominal aneurysm. PMID- 2562992 TI - Regulation of poly(A) site selection in adenovirus. AB - We have investigated the mechanisms involved in the early-to-late RNA-processing switch which regulates the mRNA species generated from the adenovirus major late transcription unit (MLTU). In particular, polyadenylation choice mechanisms were characterized by using a reconstructed adenovirus E1A gene as a site for insertion of MLTU poly(A) regulation signals (L1 and L3). Adenovirus constructs containing the variant poly(A) recognition elements were used to compare E1A poly(A) signal utilization with wild-type MLTU (L1 to L5) utilization. In both early and late stages of infection, either polyadenylation site (L1 or L3) is capable of being utilized when presented as the only operational poly(A) site. In an early infection, a virus which contains multiple elements presented in tandem (L13) uses the first poly(A) site, L1, preferentially (ratio of L1 to L3, 8:1) in both E1A and MLTU loci. Transcription termination is not involved in restricting the utilization of the downstream L3 site. In a late infection, when each of the five MLTU poly(A) sites is used, a switch also occurs for the E1AL13 construct, with utilization of both the L1 and L3 poly(A) sites. The switch from early to late was not the result of altered processing factors in the late infection, as demonstrated by superinfecting the E1AL13 construct into cells which had already entered a late stage of infection. The superinfecting virus gave an L1-only phenotype; therefore, a cis mechanism is involved in adenovirus poly(A) regulation. PMID- 2562994 TI - Discordant expression of terminal transferase and T cell receptor beta chain in fetal and pediatric thymocytes. AB - Human thymuses at different ages of development were analyzed for TdT+/beta F1+ double-stained cells. beta F1 is a mAb which recognizes a "hidden" framework determinant on the beta chain of the T cell receptor (TCR). We have found that TCR beta chains appear early during thymic ontogenesis and are detectable by 15 weeks of gestation in cells that are TdT-. Paradoxically, the immature CD2- large thymic blasts in late fetal development and in infants are TdT+ but beta chain negative. These data are compatible with the notion that TdT acts as somatic mutagen on TCR beta genes starting only after 20 weeks of gestation. The beta chain proteins which appear early during thymic ontogenesis might reflect expression of incompletely rearranged (DJ), unrearranged, or fully rearranged TCR beta genes in an early wave of thymocyte clonal (beta) diversification, that is, in contrast to later development, independent of TdT. PMID- 2562995 TI - The Stroke Council and the Young Investigator award. PMID- 2562996 TI - The effects of an internal analgesic formulary restriction on Medicaid drug expenditures in Wisconsin. AB - The effects of removing propoxyphene napsylate products from the Wisconsin Medicaid drug program formulary were examined. Internal analgesic expenditures and usage data for 3-month periods before and after the removal were compared (April through June 1984 versus the same period in 1985). After adjusting for price and reimbursement changes between the two study periods, overall expenditures were slightly higher after removal of these products. Expenditures per recipient, prescription, and unit all increased, as did the number of prescriptions per recipient. Expenditures, prescriptions, and recipients increased more for propoxyphene hydrochloride products as substitutes for propoxyphene napsylate products than for products in any other category. Increases also occurred for nonsteroidal anti-inflammatory products, suggesting they may have been chosen as replacement therapy. The proportion of napsylate prescriptions converted to hydrochloride prescriptions was larger for institutional patients than for noninstitutional patients. Although program expenditures did not decrease, as intended by the formulary change, other qualitative outcomes also should be considered, such as any therapeutic advantages the replacement products may have had for the patients. PMID- 2562998 TI - [Do beta blockaders prevent myocardial infarction?]. PMID- 2562997 TI - Nutrient balance and obesity: an approach to control of food intake in humans. AB - This article has examined the regulated systems that control nutrient balance. From this analysis, the following conclusions may be suggested: 1. Each nutrient is regulated separately in a feedback system. 2. The control of glucose is regulated by the size of the glycogen stores; the size of the fat depots, by the rate of hepatic fatty acid oxidation; and protein, by the size of the protein depots. 3. Obesity can occur as a result of hyperphagia or from repartitioning the deposition of nutrients. In either case, there is a relative or absolute reduction in the activity of the sympathetic nervous system, requiring adequate levels of circulating corticosteroids. PMID- 2562999 TI - [Secondary prevention after myocardial infarction--beta blockade and acetylsalicylic acid is the basic therapy]. PMID- 2563000 TI - Multicentre randomised clinical trial of chorion villus sampling and amniocentesis. First report. Canadian Collaborative CVS-Amniocentesis Clinical Trial Group. AB - 2787 women who were aged 35 years or more at expected date of delivery were randomised to chorionic villus sampling (CVS) at 9-12 weeks gestation or amniocentesis at 15-17 weeks for the detection of a chromosomal abnormality in the fetus. 396 women were excluded after randomisation because of a non-viable fetus, a multiple pregnancy, or infection or because the pregnancy was too far advanced (more than 12 completed weeks). Among all women eligible at the time of the first study procedure there were 89/1169 (7.6%, 95% confidence interval [CI] 6.2-9.3%) total losses (spontaneous and induced abortions and late losses) in the CVS group and 82/1174 (7.0%, CI 5.6-8.6%) in the amniocentesis group for an excess of 0.6% for those women undergoing CVS, with a tendency to later losses in the CVS group. Approximate 95% CIs indicate that this difference is most unlikely to be greater than 2.7%. Mean birthweights for each week of gestation were similar in both groups, with no evidence of excess small-for-gestational age babies in the CVS group. The proportion of preterm births were similar in both groups. Perinatal mortality was greater in the CVS group, the greatest imbalance being beyond 28 weeks. Preliminary analysis has not disclosed an obvious recurrent event in the CVS group which might explain a cause-and-effect relationship for these late fetal losses. There is no evidence of any excess intrauterine growth retardation babies in the CVS group. Maternal morbidity was similar in both groups. 103/1037 (9.9%) women eligible for CVS had a further amniocentesis; 32 of these second procedures were needed to complete the cytogenetic diagnosis. More problems such as confined mosaicism and maternal cell contamination occurred in the interpretation of the CVS samples, but these were clarified by amniocentesis when appropriate. PMID- 2563001 TI - Removal of anti-HLA antibodies by extracorporeal immunoadsorption to enable renal transplantation. AB - 10 highly and persistently sensitised patients awaiting renal transplantation underwent extracorporeal immunoadsorption to remove anti-HLA antibodies. 7 patients have since received transplants. Only 1 allograft has been lost because of rejection, and there have been no serious side-effects attributable to treatment. Extracorporeal immunoadsorption may therefore be of considerable value in the management of highly sensitised patients. PMID- 2563002 TI - Hearing impairment in patients with antibody production against Borrelia burgdorferi antigen. AB - This study aimed to evaluate the extent to which hearing disorders may be a result of tick-borne Borrelia burgdorferi infection. 98 patients with different patterns of hearing dysfunction were studied. The patients had a history of sudden hearing loss, disorders similar to Meniere's disease, or hearing loss in combination with acute facial palsy or with vertigo. Serum antibodies against the B burgdorferi antigen were determined during the acute and convalescent periods. 17 patients (17%) showed serological evidence of borreliosis (reciprocal titre of 320 or above). All but 3 of these patients also had vertigo and 3 subjects had peripheral facial palsy. All the 17 patients were treated with high doses of intravenous benzylpenicillin. The hearing of 5 patients improved on treatment. Although the specificity of antibody production against borrelia antigen has not been completely clarified, it is concluded that repeated serological examinations are worthwhile in patients with unexplained hearing disorders. PMID- 2563003 TI - Low-dose interleukin-2 induces systemic immune responses against HBsAg in immunodeficient non-responders to hepatitis B vaccination. AB - A metabolic monocyte defect appears to correlate with non-responsiveness to hepatitis B vaccine in many patients on haemodialysis. This defect prevents production of interleukin-2 during T-cell activation after antigen contact. Receptors for interleukin-2 are, however, expressed in greater numbers than in healthy subjects or uraemic responders to hepatitis B vaccination. In this study, ten uraemic patients, previous non-responders to vaccination against hepatitis B, were revaccinated with the same vaccine combined with one intramuscular injection (2.5 x 10(5) U) of natural human interleukin-2. Systemic production of antibodies against hepatitis B surface antigen was initiated in those immunodeficient patients whose cellular interleukin-2 receptor levels were found to be enhanced. PMID- 2563004 TI - Cancer risks of oral contraception. PMID- 2563005 TI - An arm and a leg. PMID- 2563006 TI - Atlantoaxial instability in Down syndrome. PMID- 2563008 TI - Blastomycosis--one disease or two? PMID- 2563007 TI - Making light work of Bowen's disease. PMID- 2563009 TI - Uncommon causes of stroke. PMID- 2563010 TI - Dietary fatty acids and inflammatory skin disease. PMID- 2563011 TI - Effect of measles, mumps, rubella vaccination on pattern of encephalitis in children. AB - 462 patients (269 males, 193 females, aged from 1 month to 16 years) with encephalitis were treated at the Children's Hospital, University of Helsinki, over a 20-year period. The incidence of encephalitis was 8.3/100,000 child-years (range 19.8 in 1974 to 2.5 in 1985 and 1986). The organisms most commonly associated with encephalitis in children were mumps, measles, and varicella viruses, and Mycoplasma pneumoniae. After the start of the nationwide measles, parotitis, and rubella (MPR) vaccination programme in 1982 in Finland, encephalitides associated with these viruses seem to have totally vanished. Currently the pathogens most often associated with childhood encephalitides are varicella-zoster, M pneumoniae, and enteroviruses. 3% of the 462 patients died from their illness, and 7% became severely damaged, with the poorest outcome occurring after multiple infections, and herpes simplex virus, cytomegalovirus or M pneumoniae infections. The decline in the total number of cases of encephalitis was not accompanied by a decrease in number of patients with a poor outcome. Patients with treatable encephalitides due, for example, to M pneumoniae and herpes viruses, need prompt attention. PMID- 2563012 TI - Outbreak of trichothecene mycotoxicosis associated with consumption of mould damaged wheat production in Kashmir Valley, India. AB - During June to September, 1987, there were reports that a considerable segment of the population of Kashmir Valley, India, were affected by a gastrointestinal disorder. Epidemiological investigations and laboratory based studies indicated that the outbreak was associated with the consumption of bread made from mould damaged wheat. The disease was not age or sex specific. Evidence of mould damage of wheat consisted of the presence of moulds (such as Fusarium sp, Aspergillus sp), and varying quantities of trichothecene mycotoxins (such as deoxynivalenol, nivalenol, acetyldeoxynivalenol, T-2 toxin) in samples tested. The symptoms were reproduced in dogs fed extracts of contaminated samples. The finding that trichothecene mycotoxins, especially deoxynivalenol trichothecene, cause symptoms in man emphasizes the need for a reappraisal of its safety limits in food. PMID- 2563013 TI - Preventing iron deficiency in at-risk communities. PMID- 2563014 TI - Human herpesvirus 6 in lymph nodes. PMID- 2563015 TI - New T-lymphotropic human herpesviruses. PMID- 2563016 TI - Oral contraceptives and malignancies of genital tract. PMID- 2563017 TI - Anti-HIV effects of alpha-interferon. PMID- 2563018 TI - Homosexual role separation and spread of AIDS. PMID- 2563019 TI - Nasopharyngeal lymphatic tissue hypertrophy in HIV-infected patients. PMID- 2563020 TI - Early diagnosis of Pneumocystis carinii infection. PMID- 2563021 TI - Endoscopic sphincterotomy in acute gallstone pancreatitis. PMID- 2563022 TI - Hepatitis A and frozen raspberries. PMID- 2563023 TI - Prevention of hepatitis B transmission at birth. PMID- 2563024 TI - Low prevalence of hepatitis B in mental handicap hospital. PMID- 2563025 TI - Hepatitis D (delta) infection in south-east London. PMID- 2563026 TI - Material immune responses and recurrent miscarriage. PMID- 2563027 TI - Multiple sclerosis or blood-brain barrier disease. PMID- 2563028 TI - DNA probes in differential diagnosis of Becker muscular dystrophy and spinal muscular atrophy. PMID- 2563029 TI - Absence of dystrophin in Becker muscular dystrophy. PMID- 2563030 TI - False allegations of child abuse. PMID- 2563031 TI - Priorities in treatment. PMID- 2563032 TI - Criticism of IPPNW. PMID- 2563033 TI - Orchidopexy at or after seven years of age. PMID- 2563034 TI - Benign type of malignant syndrome. PMID- 2563035 TI - Investigating young patients with dyspepsia. PMID- 2563036 TI - Clonogenic myeloma cells and chemotherapy. PMID- 2563037 TI - Insertion in prion protein gene in familial Creutzfeldt-Jakob disease. PMID- 2563038 TI - Prevention of NSAID-induced gastric ulcer with prostaglandin analogues. PMID- 2563039 TI - Anti-neutrophil cytoplasmic antibody and subglottic stenosis. PMID- 2563040 TI - Neuropsychological principles applied to rehabilitation of a stroke patient. PMID- 2563041 TI - Haemorrhoids and defaecatory habits. PMID- 2563042 TI - Brain damage due to blocked tracheostomy tube: a 12-year saga. PMID- 2563044 TI - Why so many fractured hips? PMID- 2563043 TI - Food irradiation. PMID- 2563045 TI - Controlled trial of transdermal nicotine patch in tobacco withdrawal. AB - A transdermal nicotine patch, which delivers 0.7 mg/cm2 per 24 h and is available in sizes of 10, 20, and 30 cm2 was tested in subjects from 21 general medical practices in a 3-month, placebo-controlled randomised double-blind study. The nicotine group (n = 100) and the placebo group (n = 99) were similar at entry. Participants who smoked more than 20 cigarettes a day were treated with the 30 cm2 patch and the others with the 20 cm2 patch. When abstinence, defined as smoking 0-3 cigarettes per week and verified by CO measurement, was achieved, the next smallest patch was applied. After 1, 2, and 3 months of treatment 41, 36, and 36%, respectively, in the nicotine group were abstinent. The corresponding figures in the placebo group were 19, 20, and 23%. The differences were significant for all 3 months. Body weight did not increase in the nicotine group, but in the placebo group the mean increase was 4.4 kg. Craving and withdrawal symptoms decreased more with nicotine substitution for cigarettes. The patches were generally well tolerated, although 25% of subjects in the nicotine group and 13% in the placebo group had transient local erythema after application of the patch; 5 members of the nicotine group withdrew because of poor cutaneous tolerance. PMID- 2563046 TI - Adjuvant tamoxifen in early breast cancer: occurrence of new primary cancers. AB - The frequency of new primary cancers was studied in 1846 postmenopausal patients included in a randomised trial of tamoxifen as an adjunct to primary surgery for early breast cancer. The median follow-up was 4.5 years (range 0.5-10.5 years). The number of new cancers in the tamoxifen group (n = 57) did not differ significantly from that in the control group (n = 70). However, in tamoxifen patients second breast cancers occurred less often and endometrial cancer occurred more often than in the controls. The increase in endometrial cancers was probably related to the agonistic oestrogenic effects of tamoxifen and was most pronounced in those treated for over 2 years. PMID- 2563047 TI - Diagnostic criteria for adult respiratory distress syndrome: time for reappraisal. AB - To assess whether current diagnostic criteria for the adult respiratory distress syndrome (ARDS) may limit research to patients with a range of severity of respiratory failure and risk factors for ARDS were studied. In 28 patients ARDS did not develop; in 9 it did; and the other 13 met the diagnostic criteria at their first assessment. Plasma concentration of neutrophil elastase-alpha 1 antitrypsin complex was higher in all groups than in controls. For all patients, plasma elastase-complex was related to worsening hypoxia and to increased bronchoalveolar lavage protein content. Elastase-complex was present in all bronchoalveolar lavage samples and was related to protein content and differential neutrophil counts. Patients at risk of or with ARDS had a spectrum of respiratory failure to which intravascular and intra-alveolar neutrophil elastase release and capillary permeability were related. This suggests that the state recognised as ARDS is not a distinct pathophysiological entity. PMID- 2563048 TI - Human papilloma virus infection and skin cancer in renal allograft recipients. AB - 202 renal allograft recipients in south-east Scotland, who had received transplants between 1965 and 1986, were monitored over 3 years (1984-87) for the presence of warts, keratoses, and skin cancers. 77% of 69 patients with graft survival of more than 5 years had viral warts, 38% had keratoses, and 12% had skin cancers, whereas of the 133 with graft survival of less than 5 years 20% had warts, 17% had keratoses, and 1.5% had skin cancers. The ratio of squamous cell carcinoma to basal cell carcinoma was 15:1. Most viral warts showed significant epidermal dysplasia, and keratoses and squamous cell carcinomas had signs of human papilloma virus infection. 15 (60%) of 25 squamous cell carcinomas contained HPV5/8 DNA and 1 contained HPV4 DNA--HPV5/8 DNA was detected in skin lesions of recipients with cancers significantly more often than in those matched for duration and type of immunosuppression with nonmalignant skin lesions. The findings suggest a role for HPV5/8 in the aetiology of squamous cell carcinoma in renal allograft recipients. PMID- 2563049 TI - Intestinal-type gastric carcinoma and colonic carcinoma: a common pathogenesis? AB - Intestinal-type gastric carcinoma and colorectal carcinoma have similar histological appearances. In the early stages of both disorders, undifferentiated cells capable of division are found on mucosal surfaces. These undifferentiated surface cells may be important in the pathogenesis of both neoplasms. PMID- 2563050 TI - SPECT and PET in epilepsy. PMID- 2563051 TI - Management of alloimmune neonatal thrombocytopenia. PMID- 2563052 TI - The dangers of not going to bed. PMID- 2563053 TI - Plastic or paraffin? PMID- 2563054 TI - ARDS times. PMID- 2563055 TI - Impact of an essential drugs programme on availability and rational use of drugs. AB - Availability and rational use of drugs was assessed in a random sample of 19 peripheral health units in two governorates in Democratic Yemen in which an essential drugs programme has been operational for the past few years. Findings were compared with those from seven health units in one governorate in which no such programme had been started. On average, 27 essential drugs were available in the programme area, compared with 17 in the control area. Programme areas carried on average 1 non-essential drug, compared with 17 in control areas. Average stock was adequate for 4 weeks in programme areas and for 1 week in control areas. Health workers in the programme area scored slightly, but not significantly, better in a test on theoretical knowledge on rational drug use. However, programme areas differed considerably from control areas in patterns of drug use, with fewer injections (24.8% vs 57.8% of prescriptions) and fewer antibiotics (46.3% vs 66.8%) being prescribed in programme areas, which also had fewer drugs per prescription (1.5 vs 2.4). The programme has significantly improved the availability and rational use of essential drugs in peripheral health units. PMID- 2563057 TI - Abdominal incision: decision or indecision? PMID- 2563056 TI - Is snoring a cause of vascular disease? An epidemiological review. AB - Eight studies that examined the relation between snoring and vascular disease were identified. The prevalence of habitual snoring, measured by questionnaire or interview, varied from 3% to 29% of adults and was dependent on age, sex, obesity, and smoking habit. In men, habitual snoring was associated with hypertension and ischaemic heart disease, with adjusted relative risks in the range 1.3-2.0. For women, only one study provided adjusted estimates of relative risk, which were 2.8 for hypertension and 1.2 for angina. Adequately adjusted relative risks for cerebrovascular disease have not been reported, but unadjusted estimates varied from 1.6 to 10.3. These studies had several limitations, including the lack of a standard definition of snoring, the use of unvalidated questionnaires, and failure to account for confounding variables and the possibility of reporting bias. Only one study was prospective. Epidemiological criteria for a causal association between snoring and vascular disease have not been satisfied. The apparent excess risk is probably due to the consequences of sleep apnoea rather than snoring itself. PMID- 2563058 TI - Policies, pills, and political will: a critique of policies to improve the health status of ethnic minorities. PMID- 2563059 TI - Increased vertebral bone density in heavy drinkers of mineral water rich in fluoride. PMID- 2563060 TI - Duodenal ulcer "epidemic" in a pathology department. PMID- 2563062 TI - Allele losses in breast cancer. PMID- 2563061 TI - Enoximone as alternative to mechanical support while awaiting cardiac transplantation. PMID- 2563063 TI - Mupirocin for eradication of nasal carriage of staphylococci. PMID- 2563064 TI - Percutaneous laser coronary angioplasty without balloon angioplasty. PMID- 2563065 TI - Flumazenil and hepatic encephalopathy. PMID- 2563066 TI - Foscarnet infusion at home. PMID- 2563067 TI - Interferon-alpha plus zidovudine in HIV infection. PMID- 2563068 TI - Changing to human insulin. PMID- 2563069 TI - Abnormal glutamatergic mechanisms and branched-chain aminoacids in amyotrophic lateral sclerosis. PMID- 2563070 TI - Anion gap. PMID- 2563071 TI - Chlamydia TWAR and acute myocardial infarction. PMID- 2563072 TI - Urine test for adenylosuccinase deficiency in autistic children. PMID- 2563073 TI - Treating ascites. PMID- 2563074 TI - Teachers, stress, and mortality. PMID- 2563075 TI - Autoclaves for rural health facilities. PMID- 2563076 TI - Gastrobronchial reflux in patients on artificial ventilation and antacid therapy. PMID- 2563077 TI - Cystic fibrosis with and without meconium ileus. PMID- 2563078 TI - Surgery for temporal lobe epilepsy. PMID- 2563079 TI - The Bamako initiative. PMID- 2563080 TI - A patient voice at the General Medical Council. PMID- 2563081 TI - Bezafibrate-induced headache. PMID- 2563082 TI - Fluconazole in renal candidosis. PMID- 2563083 TI - Treatment of ulcerative colitis by implantation of normal colonic flora. PMID- 2563084 TI - Antimycobacterial therapy ineffective in Crohn's disease after a year. PMID- 2563085 TI - Stokes-Adams attacks with migraine. PMID- 2563086 TI - Clinically and neuropathologically distinct form of dementia in the elderly. PMID- 2563087 TI - Near-simultaneous adenocarcinoma of pancreas in husband and wife. PMID- 2563088 TI - Acute psychosis with carbamazepine and sodium valproate. PMID- 2563089 TI - Measles, mumps, and rubella immunisation. PMID- 2563090 TI - Erythromycin resistance in streptococci. PMID- 2563091 TI - Hepatotoxicity of actinomycin-D. PMID- 2563092 TI - Encephalopathy associated with fat embolism induced by solvent for cyclosporin. PMID- 2563093 TI - Continuous positives airway pressure in AIDS. PMID- 2563094 TI - No-fault compensation. PMID- 2563095 TI - AIDS in the UK and world wide. PMID- 2563096 TI - Placebo-controlled, randomised trial of warfarin and aspirin for prevention of thromboembolic complications in chronic atrial fibrillation. The Copenhagen AFASAK study. AB - From November, 1985, to June, 1988, 1007 outpatients with chronic non-rheumatic atrial fibrillation (AF) entered a randomised trial; 335 received anticoagulation with warfarin openly, and in a double-blind study 336 received aspirin 75 mg once daily and 336 placebo. Each patient was followed up for 2 years or until termination of the trial. The primary endpoint was a thromboembolic complication (stroke, transient cerebral ischaemic attack, or embolic complications to the viscera and extremities). The secondary endpoint was death. The incidence of thromboembolic complications and vascular mortality were significantly lower in the warfarin group than in the aspirin and placebo groups, which did not differ significantly. 5 patients on warfarin had thromboembolic complications compared with 20 patients on aspirin and 21 on placebo. 21 patients on warfarin were withdrawn because of non-fatal bleeding complications compared with 2 on aspirin and none on placebo. Thus, anticoagulation therapy with warfarin can be recommended to prevent thromboembolic complications in patients with chronic non rheumatic AF. PMID- 2563097 TI - Case-control study of serum immunoglobulin-E antibodies reactive with soybean in epidemic asthma. AB - Since 1981, twenty-six asthma outbreaks have been identified in Barcelona, all coinciding with the unloading of soybean in the harbour. Serum from patients with epidemic asthma and individually matched controls with non-epidemic asthma was assayed for immunoglobulin-E (IgE) antibodies against soybean antigens by means of a radioallergosorbent test. In 64 of 86 cases (74.4%) there was a reaction with commercial soybean antigen extracts, compared with only 4 of the 86 controls (4.6%) (odds ratio = 61; lower 95% confidence limit = 8.1). The statistical significance was greater for reactions with extracts of soybean dust taken from Barcelona harbour (odds ratio, unquantifiably high; lower 95% confidence limit = 11.7). No other serological covariate (total serum IgE levels or specific IgE levels against the commonest airborne allergens or legumes) confounded the association between serum anti-soybean IgE antibodies and epidemic asthma. These results support a causal relation between the release of dust during unloading of soybean at the harbour and the occurrence of asthma outbreaks, suggesting an underlying allergic mechanism. PMID- 2563099 TI - Possible role for vitamin D in controlling breast cancer cell proliferation. AB - By means of an immunocytochemical method the 1.25-dihydroxyvitamin D [1.25(OH)2D] receptor status of tumours from 136 patients with primary carcinoma of the breast was determined. Patients with receptor-positive tumours had significantly longer disease-free survival than those with receptor-negative tumours (Chi2 = 4.01, p less than 0.05). 1.25(OH)2D3 inhibits the proliferation of several established human breast cancer cell lines in vitro. Effects of 1.25(OH)2D3 on breast tumour growth in vitro were assessed by means of the nitrosomethylurea-induced rat mammary tumour model of hormone-responsive breast cancer. Treatment of tumour bearing animals with 0.1 microgram of the synthetic analogue, 1 alpha hydroxyvitamin D3, three times weekly produced significant inhibition of tumour progression. Taken together, these studies suggest that the levels of 1.25(OH)2D occurring in vivo may exert an inhibitory effect on receptor-positive tumours. Further studies are required to evaluate the role of vitamin D metabolites in the treatment of human malignant disease. PMID- 2563098 TI - Lone bilateral blindness: a transient ischaemic attack. AB - In the Oxfordshire Community Stroke Project 14 patients were notified with lone bilateral blindness, defined as rapid onset of dimming or loss of vision over all of both visual fields simultaneously, lasting under 24 hours, without associated symptoms of focal cerebral ischaemia, epilepsy, or reduction in consciousness. The age of these patients was close to that of the 184 patients who presented with transient ischaemic attacks and they had a similar high prevalence of vascular risk factors. During a mean follow-up of 2.4 years, 5 of the 14 had a first-ever stroke (0.31 expected). In view of their 16 times (95% CI 7-39 times) excess risk of stroke such patients should be included, for practical purposes, under the diagnostic heading of transient ischaemic attack. PMID- 2563100 TI - Gene therapy. PMID- 2563101 TI - Herpes simplex virus latency. PMID- 2563102 TI - Investigation of rectal bleeding. PMID- 2563103 TI - Type III collagen deficiency. PMID- 2563104 TI - Surgery for pulmonary emboli? PMID- 2563105 TI - Sleeping position and infant bedding may predispose to hyperthermia and the sudden infant death syndrome. AB - Southern New Zealnd has one of the highest postneonatal mortality rates in the developed world (8.1/1000 livebirths) and 77% of these deaths are attributed to the sudden infant death syndrome (SIDS). Both hyperthermia and sleeping position have previously been implicated in SIDS. A theoretical model to estimate the thermal balance of infants used here shows that the head, and particularly the face, becomes the main route for heat loss when thick clothing and bedding are used. This thermoregulatory role could be compromised by the prone sleeping position. It is postulated that particular cultural combinations of infant care practices (sleeping position, clothing, bedding, and room heating) may facilitate hyperthermia and explain widely disparate rates of SIDS in different countries and ethnic groups. PMID- 2563106 TI - Ciguatera in the Pacific: a link with military activities. AB - Ciguatera fish poisoning is widespread in the Pacific. Outbreaks and the rise in incidence of the disease are related largely to military activities that disturb coral reef ecology. Nuclear test explosions and the setting up of the infrastructure for these tests are major components of such military activity. PMID- 2563107 TI - Hepatitis B infections after gynaecological surgery. AB - When acute hepatitis B developed in 3 patients who had had gynaecological surgery, the surgeon was found to be a carrier of hepatitis B e antigen. Of 268 patients operated on by this surgeon in one hospital, 247 were screened for markers of recent or current hepatitis B. 22 (9%) had such markers, associated with symptoms in 5. The operations carrying greatest risk of infection were hysterectomy (10/42) and caesarean section (10/51). These findings strengthen the case for vaccination of all surgeons and medical students against hepatitis B. PMID- 2563109 TI - Immunosuppression for membranous nephropathy. PMID- 2563108 TI - A "significant" stenosis: thirty years on. PMID- 2563110 TI - Investigating young patients with dyspepsia. PMID- 2563111 TI - Misoprostol and ulcer prophylaxis. PMID- 2563112 TI - Laboratory confirmation of Cortinarius poisoning. PMID- 2563113 TI - Paracetamol and peptic ulcer. PMID- 2563114 TI - 99mTc-HMPAO washout in prognosis of stroke. PMID- 2563115 TI - Gross regional cerebral hypofunction with normal CT scan in Creutzfeldt-Jakob disease. PMID- 2563116 TI - Fulminant hepatitis due to cyproterone acetate. PMID- 2563117 TI - Startle disease: an avoidable cause of sudden infant death. PMID- 2563118 TI - Ganciclovir for cytomegalovirus infection in renal transplant recipients. PMID- 2563119 TI - Human herpesvirus-6 and cross-reactivity with other herpesviruses. PMID- 2563120 TI - Adrenaline/hypertension hypothesis. PMID- 2563121 TI - Destruction of Listeria monocytogenes during microwave cooking. PMID- 2563122 TI - Adenosine and hypertrophic cardiomyopathy. PMID- 2563123 TI - Imaging patients with suspected acoustic neuroma. PMID- 2563124 TI - Multiple primary malignant neoplasms in patients with adult T-cell leukaemia. PMID- 2563125 TI - Destitution at the festive season. PMID- 2563126 TI - Drug advertisements in developing countries. PMID- 2563127 TI - Mianserin. PMID- 2563128 TI - Revising RAWP. PMID- 2563129 TI - Blastocystis hominis chronic diarrhoea in AIDS patients. PMID- 2563130 TI - Neuroleptic malignant syndrome. PMID- 2563131 TI - Aerosolised pentamidine. PMID- 2563132 TI - Chest pain after intravenous corticotropin-releasing hormone. PMID- 2563133 TI - Exercise oximetry for early diagnosis of Pneumocystis carinii pneumonia. PMID- 2563134 TI - Monoclonal antibody (HML-1) labelling of T-cell lymphomas. PMID- 2563135 TI - Hypothyroidism and goitre during interleukin-2 therapy without LAK cells. PMID- 2563136 TI - Alcohol and the U-shaped curve. PMID- 2563137 TI - Hairloss and scaling with proguanil. PMID- 2563138 TI - Hereditary elliptocytosis in Africa. PMID- 2563139 TI - Self-injury, regurgitation, and antiemetics. PMID- 2563140 TI - Unusual childhood uveitis in Tanzania. PMID- 2563141 TI - Enalapril pharmacokinetics in diabetic patients. PMID- 2563142 TI - Diabetic nephropathy and enalapril. PMID- 2563143 TI - Yellow fever virus activity--Trinidad and Tobago. PMID- 2563144 TI - In vitro induction of chromosome damage by sulphasalazine in human lymphocytes. AB - Two different endpoints, sister-chromatid exchange and micronucleus induction, were measured in human peripheral blood lymphocytes stimulated to divide in short term in vitro cultures. The cultures were exposed to sulphasalazine and 6 of its metabolites for 72 h in the absence of any exogenous metabolic activation system. Analysis of the sister-chromatid exchange and micronuclei frequencies clearly indicates that sulphasalazine itself is capable of inducing both sister-chromatid exchange and micronuclei while sulphapyridine and its acetylated metabolites only induce sister-chromatid exchange. 5-Aminosalicylic acid, the therapeutic moiety of sulphasalazine, and its acetylated metabolite did not induce either sister chromatid exchange or micronuclei at the concentrations tested. The data from these in vitro experiments are discussed in relation to the previously reported elevations in sister-chromatid exchange and micronucleus frequencies in inflammatory bowel disease patients receiving sulphasalazine therapy. PMID- 2563146 TI - The use of a DNA probe for the differentiation of rodent malaria strains and species. AB - A clone, PCsv4, derived from a partial genomic library of the rodent malaria Plasmodium chabaudi chabaudi AS strain, contains an insert which when used as a probe at low stringency in Southern blotting of genomic DNA from a variety of strains, subspecies and species of rodent malaria parasites, results in a pattern of hybridisation which is specific to each of the DNA samples used, thus providing an accurate method to define a rodent malaria line. The insert only hybridises to DNA derived from malaria parasites of rodent species. The insert also hybridises to a small number of RNA transcripts. PMID- 2563145 TI - Different allele frequencies in Trypanosoma brucei brucei and Trypanosoma brucei gambiense populations. AB - Restriction fragment length polymorphism (RFLP) has been analysed in Trypanosoma brucei DNA following hybridization with different DNA probes. This polymorphism seems to be due to allelic variation, and not to variation between sequence duplicates, since the genomic environment of the probed polymorphic fragments is conserved over considerable distances. In an analysis of 35 non-gambiense stocks, we found different combinations of homozygotes and heterozygotes for the four RFLP probes used, in keeping with previous observations that genetic reassortment occurs in T. b. brucei. Moreover, the non-gambiense populations from West and East Africa can be differentiated according to their characteristic allele frequencies. In sharp contrast, we found that the 49 T. b. gambiense stocks, analysed with the same probes, share the same single allelic combination and are all homozygous for each one of the four markers. This characteristic gambiense allele combination is very common among Western non-gambiense isolates, but rare or absent among Eastern ones. Two stocks isolated from man in West Africa turned out to be non-gambiense by all molecular criteria examined, including total nuclear DNA content. Taken together, these observations suggest that human serum resistant variants may appear among the West African T. b. brucei population, and that T. b. gambiense evolved from one of these resistant variants as a man adapted subspecies that became genetically isolated from the rest of the West African trypanosome population. PMID- 2563147 TI - Cooling of metered-dose inhalers decreases pressure output from canisters. PMID- 2563149 TI - GABAA responses in hippocampal neurons are potentiated by glutamate. AB - In the mammalian cortex, glutamate and gamma-aminobutyric acid (GABA) are the principal transmitters mediating excitatory and inhibitory synaptic events. Glutamate activates cation conductances that lead to membrane depolarization whereas GABA controls chloride conductances that produce hyperpolarization. Here we report that the GABAA-activated conductance in hippocampal pyramidal cells is enhanced by glutamate at concentrations below that required for its excitatory action. The GABA-potentiating effect can be induced, with comparable potency, by several glutamate analogues such as quisqualate, N-methyl-D-aspartate (NMDA), kainate and, surprisingly, by D-2-amino-5-phosphonovalerate (APV), an antagonist for NMDA receptors. Data from dose-response curves show that glutamate enhances the GABAA conductance without significantly changing GABA binding affinity. The low concentration of glutamate needed to enhance GABAA responses raises the possibility that glutamate modulates the strength of GABA-mediated transmission in the cortex. PMID- 2563148 TI - Birth of the D-E-A-D box. PMID- 2563150 TI - Nuclear transcriptional activity of the tobacco plastid psbA promoter. AB - The plastid psbA promoter of tobacco was used with the aim to construct plastid specific marker genes. Upon transfer to the tobacco nuclear genome the plastid promoter fragment appeared to specify a messenger RNA. Placed 5' to the bar or nptII coding sequences the level of expression is sufficient to obtain a selectable phenotype. The transcription start site in the nucleus is site specific and is located 4 nucleotides downstream relative to the start site used in plastids. Translational fusions of the psbA coding sequence with the nptII and bar coding sequences revealed that the psbA leader sequence and the psbA translation start codon, being the second ATG codon, are recognized by the plant cytoplasmic translation apparatus. A promoter cassette utilisable in both E. coli and tobacco, was obtained by placing the CaMV 35S enhancer 5' to the psbA promoter. PMID- 2563151 TI - Cis-acting regulatory elements controlling temporal and organ-specific activity of nopaline synthase promoter. AB - Regulatory elements controlling temporal and organ-specific expression of the nopaline (nos) gene were identified by analyzing deletion mutants of the promoter. As observed in cultured cells, the TATA box element was required for efficient promoter function and the 29 bp upstream promoter region between -130 and -101 was necessary for the nos promoter activity in various vegetative organs. This 29 bp region includes ten nucleotides of a potential Z-DNA-forming sequence (Z element) and eight nucleotides of a repeated element (b element). Duplication of b elements significantly enhanced the promoter strength, revealing the importance of the element in all plant organs. Unlike the results in the cultured cells, however, deletion of the b element or CCAAT box region completely inactivated the promoter function in regenerated organs. Therefore, it appears that transcription initiation is more tightly controlled in differentiated plant cells than in cultured cells. PMID- 2563152 TI - Nucleotide sequences of the two high-molecular-weight glutenin genes from the D genome of a hexaploid bread wheat, Triticum aestivum L. cv Cheyenne. PMID- 2563153 TI - Rapid RFLP screening using DNA from complete hydatidiform moles. PMID- 2563154 TI - Ava II RFLP at the insulin-like growth factor II (IGF II) locus on chromosome 11. PMID- 2563155 TI - HinfI RFLP at the human renin (REN) gene locus. PMID- 2563156 TI - Anonymous DNA segment H33 [D6S42] on chromosome 6 associated with 2 RFLPs. PMID- 2563157 TI - Three allele RFLP identified by an anonymous sequence on chromosome 2, E135 [D2S62]. PMID- 2563158 TI - Three RFLPs upstream of the low density lipoprotein receptor (LDLR) gene. PMID- 2563159 TI - A unique RFLP in the human T-cell receptor gamma constant region gene TRGC1. PMID- 2563160 TI - A Hpa I polymorphism in the human laminin B1 chain gene on 7q22. PMID- 2563161 TI - Site-directed mutagenesis of the catalytic residues Asp-52 and Glu-35 of chicken egg white lysozyme. AB - The roles of the catalytic active-site residues aspartic acid-52 and glutamic acid-35 of chicken lysozyme (EC 3.2.1.17) have been investigated by separate in vitro mutagenesis of each residue to its corresponding amide (denoted as D52N and E35Q, respectively). The mutant enzyme D52N exhibits approximately 5% of the wild type lytic activity against Micrococcus luteus cell walls, while there is no measurable activity associated with E35Q (0.1% +/- 0.1%). The measured dissociation constants for the chitotriose-enzyme complexes were 4.1 microM (D52N) and 13.4 microM (E35Q) vs. 8.6 microM for wild type, indicating that the alterations in catalytic properties may be due in part to binding effects as well as to direct catalytic participation of these residues. The mutant lysozymes have been expressed in and secreted from yeast and obtained at a level of approximately 5 mg per liter of culture by high-salt elution from the cell walls. PMID- 2563163 TI - Overexpression of amyloid precursor protein A4 (beta-amyloid) immunoreactivity in genetically transformed cells: implications for a cellular model of Alzheimer amyloidosis. AB - Among the major obstacles to clarifying molecular mechanisms involved in amyloid metabolism in Alzheimer disease has been the unavailability of laboratory models for this uniquely human disorder. The present studies were aimed at establishing genetically engineered cell lines that overexpress amyloid immunoreactivity and that may be relevant to amyloid accumulation in the Alzheimer disease brain. We used cloned amyloid cDNA that contains a region encoding A4 (beta-polypeptide) amino acids along with recently developed tumor virus vectors derived from simian virus 40 to prepare transformed cells. After transient and permanent transfection, a variety of cell types overexpressed A4 immunoreactivity that was detected by highly specific monoclonal antibodies. We observed that the use of an amyloid subdomain containing the A4 region, but lacking the sequence of a Kunitz type protease inhibitor found in amyloid precursor protein variants, was sufficient to obtain cells that overproduced an A4 epitope. The transformed cells were readily propagated in culture and may provide an experimental medium to elucidate aspects of the molecular pathogenesis of Alzheimer disease. The cellular models may also serve as tools for deriving potentially useful therapeutic agents. PMID- 2563162 TI - cDNA sequence for the alpha M subunit of the human neutrophil adherence receptor indicates homology to integrin alpha subunits. AB - The receptor on human neutrophils (polymorphonuclear leukocytes) that mediates cellular adherence consists of two noncovalently associated subunits, designated alpha M (Mac-1 alpha, Mol alpha, or CD11b; Mr, 170,000) and beta (Mac-1 beta, Mol beta, or CD18; Mr, 100,000). We isolated a cDNA clone for the human neutrophil alpha M subunit by screening a lambda gt 11 cDNA library made from chronic myelogenous leukemia neutrophils by using an affinity-purified rabbit polyclonal antibody directed against the alpha M subunit. We used this cDNA clone to obtain additional clones from cDNA libraries made from differentiated HL-60 promyelocytic leukemia cells. Together these cDNAs constitute the complete 1137 amino acid sequence for the mature human alpha M subunit protein. The deduced amino acid sequence indicates the presence of an extensive extracellular domain with three putative metal-binding regions, (i) an amino acid region that is homologous to the A domain of von Willebrand factor, (ii) a 26-amino acid hydrophobic sequence that is a potential transmembrane domain, and (iii) a 19 amino acid cytoplasmic region. The amino acid sequence for the human neutrophil alpha M subunit contains regions that are closely related to amino acid sequences of adhesion receptors belonging to the integrin family. PMID- 2563164 TI - Phenylethanolamine N-methyltransferase-containing neurons in the limbic system of the young rat. AB - Fifteen years ago epinephrine cells were shown to be present in the medulla oblongata of the rat. These cell groups (C1 and C2) were thought to supply the epinephrine innervation in the rest of the central nervous system. In this study I demonstrate the presence of epinephrine-producing neurons in the forebrain of the young rat. Neurons that are immunopositive for phenylethanolamine N methyltransferase (S-adenosyl-L-methionine:phenylethanolamine N methyltransferase, EC 2.1.1.29) are present in the central nucleus of the amygdala as well as in the bed nucleus of the stria terminalis. Neurons in the same location are also immunopositive for tyrosine hydroxylase [tyrosine 3 monooxygenase; L-tyrosine, tetrahydrobiopterine:oxygen oxidoreductase (3 hydroxylating), EC 1.14.16.2]. The phenylethanolamine N-methyltransferase immunopositivity disappears by day 35, while a small amount of tyrosine hydroxylase-positive cells still can be found in the adult. In situ hybridization reveals tyrosine hydroxylase mRNA in the above nuclei in both young and adult animals. The number of the positive cells decreases in adulthood. RNA blot hybridization analysis showed the presence of phenylethanolamine N methyltransferase mRNA in the amygdala and the bed nucleus of the stria terminalis in the young and in the adult rat brain. Neurons that are immunopositive for phenylethanolamine N-methyltransferase are also present in the human amygdala. PMID- 2563165 TI - Consequences of stochastic release of neurotransmitters for network computation in the central nervous system. AB - Neuronal membrane potentials vary continuously due largely to background synaptic noise produced by ongoing discharges in their presynaptic afferents and shaped by probabilistic factors of transmitter release. We investigated how the random activity of an identified population of interneurons with known release properties influences the performance of central cells. In stochastic models such as thermodynamic ones, the probabilistic input-output function of a formal neuron is sigmoid, having its maximal slope inversely related to a variable called "temperature." Our results indicate that, for a biological neuron, the probability that given excitatory input signals reach threshold is also sigmoid, allowing definition of a temperature that is proportional to the mean number of quanta comprising noise and can be modified by activity in the presynaptic network, a notion which could be included in neural models. By introducing uncertainty to the input-output relation of central neurons, synaptic noise could be a critical determinant of neuronal computational systems, allowing assemblies of cells to undergo continuous transitions between states. PMID- 2563166 TI - Association between a specific apolipoprotein B mutation and familial defective apolipoprotein B-100. AB - Familial defective apolipoprotein (apo) B-100 is a genetic disease that leads to hypercholesterolemia and to an increased serum concentration of low density lipoproteins that bind defectively to the apoB,E(LDL) receptor. The disorder appears to result from a mutation in the gene for apoB-100. Extensive sequence analysis of the two alleles of one subject heterozygous for the disorder has revealed a previously unreported mutation in the codon for amino acid 3500 that results in the substitution of glutamine for arginine. This same mutant allele occurs in six other, unrelated subjects and in eight affected relatives in two of these families. A partial haplotype of this mutant apoB-100 allele was constructed by sequence analysis and restriction enzyme digestion at positions where variations in the apoB-100 are known to occur. This haplotype is the same in three probands and four affected members of one family and lacks a polymorphic Xba I site whose presence has been correlated with high cholesterol levels. Thus, it appears that the mutation in the codon for amino acid 3500 (CGG----CAG), a CG mutational "hot spot," defines a minor apoB-100 allele associated with defective low density lipoproteins and hypercholesterolemia. PMID- 2563167 TI - Tissue-specific splicing mutation in acute intermittent porphyria. AB - An inherited deficiency of porphobilinogen deaminase [porphobilinogen ammonia lyase (polymerizing), EC 4.3.1.8] in humans is responsible for the autosomal dominant disease acute intermittent porphyria. Different classes of mutations have been described at the protein level suggesting that this is a heterogeneous disease. It was previously demonstrated that porphobilinogen deaminase is encoded by two distinct mRNA species expressed in a tissue-specific manner. Analysis of the genomic sequences indicated that these two mRNAs are transcribed from two promoters and only differ in their first exon. The first mutation identified in the human porphobilinogen deaminase gene is a single-base substitution (G----A) in the canonical 5' splice donor site of intron 1. This mutation leads to a particular subtype of acute intermittent porphyria characterized by the restriction of the enzymatic defect to nonerythropoietic tissues. Hybridization analysis using oligonucleotide probes after in vitro amplification of genomic DNA offers another possibility of detecting asymptomatic carriers of the mutation in affected families. PMID- 2563168 TI - Multidrug-resistance gene (P-glycoprotein) is expressed by endothelial cells at blood-brain barrier sites. AB - Endothelial cells of human capillary blood vessels at the blood-brain and other blood-tissue barrier sites express P-glycoprotein as detected by mouse monoclonal antibodies against the human multidrug-resistance gene product. This pattern of endothelial cell expression may indicate a physiological role for P-glycoprotein in regulating the entry of certain molecules into the central nervous system and other anatomic compartments, such as the testes. These tissues, which limit the access of systemic drugs, are known pharmacologic sanctuaries for metastatic cancer. P-glycoprotein expression in capillary endothelium of brain and testes and not other tissues (i.e., kidney and placenta) may in part explain this phenomenon and could have important implications in cancer chemotherapy. PMID- 2563169 TI - Identification of cellular and extracellular sites of amyloid precursor protein extracytoplasmic domain in normal and Alzheimer disease brains. AB - Information concerning the distribution of various subdomains of the amyloid precursor protein (APP) in brain may illuminate aspects of the normal metabolism of this membrane-associated protein, as well as putative abnormal processing that may occur in Alzheimer disease (AD). We prepared affinity-purified antibody, P2, against an extracytoplasmic APP site and applied it, along with monoclonal antibodies to the beta-peptide, or A4 region, in conjunction with selective cytochemical staining methods, to control and AD tissues. The following was noted: (i) in contrast to A4 epitopes, which are easily demonstrable primarily in extracellular senile plaques of AD patients, the extracytoplasmic P2 antigen was found in association with neurons, glia, and blood vessels in both normal and AD prefrontal cortex; (ii) a subset of senile plaques contained both A4 and P2 antigens; (iii) in some instances, P2 antigen occurred as an extracellular deposit in the absence of A4; (iv) the P2 antigen, but not A4, was also associated with corpora amylacea. In addition to identifying the unique cellular distribution of the APP extracytoplasmic antigen, the results support the view that a segment of this domain undergoes processing and deposition at extracellular sites, including a subset of senile plaques. PMID- 2563170 TI - The midaortic syndrome: diagnosis and treatment. PMID- 2563172 TI - Evolution of urea synthesis in vertebrates: the piscine connection. AB - Elasmobranch fishes, the coelacanth, estivating lungfish, amphibians, and mammals synthesize urea by the ornithine-urea cycle; by comparison, urea synthetic activity is generally insignificant in teleostean fishes. It is reported here that isolated liver cells of two teleost toadfishes, Opsanus beta and Opsansus tau, synthesize urea by the ornithine-urea cycle at substantial rates. Because toadfish excrete ammonia, do not use urea as an osmolyte, and have substantial levels of urease in their digestive systems, urea may serve as a transient nitrogen store, forming the basis of a nitrogen conservation shuttle system between liver and gut as in ruminants and hibernators. Toadfish synthesize urea using enzymes and subcellular distributions similar to those of elasmobranchs: glutamine-dependent carbamoyl phosphate synthethase (CPS III) and mitochondrial arginase. In contrast, mammals have CPS I (ammonia-dependent) and cytosolic arginase. Data on CPS and arginases in other fishes, including lungfishes and the coelacanth, support the hypothesis that the ornithine-urea cycle, a monophyletic trait in the vertebrates, underwent two key changes before the evolution of the extant lungfishes: a switch from CPS III to CPS I and replacement of mitochondrial arginase by a cytosolic equivalent. PMID- 2563171 TI - Effects of buried ionizable amino acids on the reduction potential of recombinant myoglobin. AB - The temperature dependences of the reduction potentials (E degrees') of wild-type human myoglobin (Mb) and three site-directed mutants have been measured by the use of thin-layer spectroelectrochemistry. Residue Val68, which is in van der Waals contact with the heme in Mb, has been replaced by Glu, Asp, and Asn. The changes in E degrees' and the standard entropy (delta S degrees') and enthalpy (delta H degrees') of reduction in the mutant proteins were determined relative to values for wild type; the change in E degrees' at 25 degrees C was about -200 millivolts for the Glu and Asp mutants, and about -80 millivolts for the Asn mutant. At pH 7.0, reduction of Fe(III) to Fe(II) in the Glu and Asp mutants is accompanied by uptake of a proton by the protein. These studies demonstrate that Mb can tolerate substitution of a buried hydrophobic group by potentially charged and polar residues and that such amino acid replacements can lead to substantial changes in the redox thermodynamics of the protein. PMID- 2563173 TI - Duplication, deletion, and polymorphism in the sex-determining region of the mouse Y chromosome. AB - The ZFY gene in the sex-determining region of the human Y chromosome encodes a "zinc-finger" protein that may be the testis-determining factor, TDF. Although the Y chromosomes of most placental mammals carry a single homolog of ZFY, the mouse Y chromosome has two homologs, both in the sex-determining (Sxr) region. Zfy-1 alone may suffice to determine maleness; Zfy-2 is dispensable, as it was deleted in an Sxr variant that retains sex-determining function but has lost other genes. Both loci mapped near the centromere of the mouse Y chromosome. The Y chromosomes of the subspecies Mus musculus musculus and M. m. domesticus were distinguishable by a Zfy-1 restriction fragment polymorphism, which can be used to study their differing interactions with autosomal sex-determining genes. PMID- 2563174 TI - Chromosome mapping and expression of a putative testis-determining gene in mouse. AB - Isolation and mapping of a mouse complementary DNA sequence (mouse Y-finger) encoding a multiple, potential zinc-binding, finger protein homologous to the candidate human testis-determining factor gene is reported. Four similar sequences were identified in Hind III-digested mouse genomic DNA. Two (7.2 and 2.0 kb) were mapped to the Y chromosome. Only the 2.0-kb fragment, however, was correlated with testis determination. Polymerase chain reaction analysis suggests both Y loci are transcribed in adult testes. A 3.6-kb fragment was mapped to the X chromosome between the T16H and T6R1 translocation breakpoints, and a fourth (6.0 kb) was mapped to chromosome 10. Hence, mYfin sequences have been duplicated several times in the mouse, although they are not duplicated in humans. PMID- 2563175 TI - Molecular cloning of genes under control of the circadian clock in Neurospora. AB - To investigate the regulation of messenger RNA abundance by circadian clocks, genomic and complementary DNA libraries were screened with complementary DNA probes enriched, by means of sequential rounds of subtractive hybridization, for sequences complementary to transcripts specific to either early morning or early evening cultures of Neurospora. Only two morning-specific genes were identified through this protocol. RNA blot analysis verified that the abundance of the transcripts arising from these genes oscillates with a period of 21.5 hours in a clock wild-type strain and 29 hours in the long-period clock mutant strain frq7. Genetic mapping through the use of restriction fragment length polymorphisms shows the two genes, ccg-1 and ccg-2, to be unlinked. These data provide a view of the extent of clock control of gene expression. PMID- 2563177 TI - A genetic polymorphism in the renin gene of Dahl rats cosegregates with blood pressure. AB - Blood pressure is influenced by multiple genetic loci whose identities are largely unknown. A restriction fragment length polymorphism (RFLP) in the renin gene was found between Dahl salt-hypertension-sensitive (S) and Dahl salt hypertension-resistant (R) rats. In an F2 population derived from crossing S and R rats, the renin RFLP cosegregated with blood pressure. One dose of the S-rat renin allele was associated with an increment in blood pressure of approximately 10 mmHg, and two doses of this allele increased blood pressure approximately 20 mmHg. From this it can be definitively concluded that in the rat the renin gene is, or is closely linked to, one of the genes regulating blood pressure. PMID- 2563176 TI - Role for excitatory amino acids in methamphetamine-induced nigrostriatal dopaminergic toxicity. AB - The systemic administration of either methamphetamine or 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine (MPTP) to experimental animals produces degenerative changes in nigrostriatal dopaminergic neurons or their axon terminals. This study was conducted to determine if excitatory amino acids, which appear to be involved in various neurodegenerative disorders, might also contribute to the dopaminergic neurotoxicity produced in mice by either methamphetamine or MPTP. MK-801, phencyclidine, and ketamine, noncompetitive antagonists of one subtype of excitatory amino acid receptor, the N-methyl-D-aspartate receptor, provided substantial protection against neurotoxicity produced by methamphetamine but not that produced by MPTP. These findings indicate that excitatory amino acids play an important role in the nigrostriatal dopaminergic damage induced by methamphetamine. PMID- 2563178 TI - Phylogeny and molecular data. PMID- 2563179 TI - Simultaneous plasma determination of floctafenin and its major metabolites by high-performance liquid chromatography: preliminary observations in children. AB - An isocratic reversed-phase ion-pair liquid chromatography with UV detection at 350 nm for the determination in human plasma of floctafenin (F) and its three main metabolites--floctafenic acid (FA), hydroxyfloctafenin (HOF), and hydroxyfloctafenic acid (HOFA)--is reported. Analytes and internal standard were extracted from acid plasma into ethyl acetate, and this organic phase was evaporated to dryness. This extraction yielded plasma drug recoveries of greater than 72%. Using 1 ml of plasma, the lower quantification limit was 0.05 microgram ml-1 with excellent linearity up to 0.8 microgram ml-1 for HOF and HOFA and up to 4.0 micrograms ml-1 for F and FA. The reproducibility and the selectivity of the method for several drugs thought likely to be administered in conjunction with F, were demonstrated. This method has been successfully applied to a pharmacokinetic study with a single 10 mg kg-1 oral dose in ten children. PMID- 2563180 TI - Hyperprealbuminemia, euthyroid hyperthyroxinemia, Zollinger-Ellison-like syndrome and hypercorticism in a pancreatic endocrine tumour. AB - Prealbumin, one of the main thyroxine transport proteins, has recently been shown to be a valuable immunohistochemical marker of neuroendocrine tumours. We report the case of a multisecretory pancreatic endocrine tumour whose prealbumin secretion was so high that it produced a peak on routine serum protein electrophoresis and induced a euthyroid hyperthyroxinemia. The maximal binding capacity of prealbumin for thyroxine was indeed markedly increased, whereas its affinity for this hormone was normal. The tumour was associated with gastric hyperacidity and hypergastrinemia thereby evoking a Zollinger-Ellison syndrome. The secretin stimulation test and gastrin tumoural immunohistochemistry were, however, negative. We suggest that the concomitant tumoural production of gastrin releasing peptide was responsible for the gastric hyperacidity and hypergastrinemia. This hormone probably also accounted for a moderate hypercorticism. PMID- 2563181 TI - Inhibition of thyrotropin-stimulated adenylate cyclase activation and growth of rat thyroid cells, FRTL-5, by immunoglobulin G from patients with primary myxedema: comparison with activities of thyrotropin-binding inhibitor immunoglobulins. AB - We studied the blocking type TSH receptor antibodies in 28 patients with primary myxedema and 21 patients with goitrous Hashimoto's thyroiditis by measuring the ability of their IgGs to inhibit TSH binding to its receptor, and to inhibit TSH stimulated cAMP increase and [3H]thymidine incorporation in a rat thyroid cell line, FRTL-5. The incidences of TSH binding inhibitor immunoglobulin, thyroid stimulation inhibiting immunoglobulin and thyroid growth inhibiting immunoglobulin in patients with primary myxedema were 54.6, 75 and 65.2%, respectively, against 14.3, 0 and 17.7%, respectively, in goitrous Hashimoto's thyroiditis. The antibodies inhibited dose-dependently not only TSH stimulated but also Graves' IgG-stimulated cAMP increase and [3H]thymidine incorporation. The TSH binding inhibitor immunoglobulin activities in patients with primary myxedema were significantly correlated with both the thyroid stimulation inhibiting immunoglobulin (r = 0.665; P less than 0.01) and the thyroid growth inhibiting immunoglobulin (r = 0.618; P less than 0.01) activity. Thirteen patients whose TSH binding inhibitor immunoglobulin activities were more than 50% had both strong thyroid stimulation inhibiting immunoglobulin (75.1-100%) and thyroid growth inhibiting immunoglobulin (57.4-100%) activities. These data suggest that the vast majority of patients with primary myxedema have potent blocking type TSH receptor antibodies. These might play a role in primary myxedema causing hypothyroidism and thyroid atrophy through inhibiting TSH stimulated cAMP generation. PMID- 2563182 TI - The influence of the pelvic nerves on anorectal motility in the cat. AB - The influence of the parasympathetic pelvic nerves on anorectal motility was studied in anaesthetized cats. Anal pressure and rectal motility were recorded by a manometric and a volumetric method, respectively. Severing of the pelvic nerves did not cause any pressure change in the anus, indicating that these nerves are not significantly tonically active. Efferent low intensity (0.05-0.5 ms, 8 V at 5 Hz) electrical stimulation of the pelvic nerves (PNS) elicited a contraction of the internal anal sphincter (IAS), while high intensity stimulation (greater than 1 ms, 8 V at 5 Hz) caused a sphincter relaxation. A rectal contraction was noted on both low and high intensity stimulation. After sectioning of the sympathetic nerves, PNS elicited a contraction in both the anus and the rectum irrespective of stimulation intensity. PNS inhibited the anal contraction elicited by simultaneous stimulation of the sympathetic nerves or noradrenaline infusion. The inhibitory anal responses to PNS were unaffected or augmented by atropine, unaffected by propranolol and abolished by hexamethonium. The excitatory anal effects of PNS were reduced or abolished by atropine and abolished by phentolamine. The rectal contraction induced by low intensity PNS was abolished by atropine or converted to a relaxation. In half of the experiments an atropine resistant rectal contraction was observed in response to high intensity PNS. The results are consistent with a pelvic nerve influence on IAS pressure through several mechanisms, including modulation of the activity in the sympathetic nerves and activation of inhibitory non-adrenergic, non-cholinergic neurons. The pelvic nerves convey both cholinergic and non-cholinergic excitatory, as well as non-adrenergic, non-cholinergic inhibitory fibres to the rectum. PMID- 2563183 TI - On pre- and/or post-junctional roles of ATP and an unknown 'substance X' as sympathetic co-transmitters in rat tail artery. PMID- 2563184 TI - Drug testing at the oral boards? PMID- 2563186 TI - Valvular prolapse and great vessel dilatation in multiple endocrine neoplasia type 2B. PMID- 2563185 TI - Hemodynamic, antiischemic, and neurohumoral effects of enoximone in patients with coronary artery disease. AB - To evaluate the risk of ischemia in 17 patients with significant coronary artery disease, the influence of enoximone was analyzed under the following conditions: (1) at rest (RC) and during exercise (ExC) under control conditions and (2) at rest (RE) and during exercise (ExE) after administration of enoximone (0.75 mg/kg, intravenously). During ExC all patients had ischemia (angina, and ST segment alterations); metabolic markers of ischemia (MMI) increased, as did the mean pulmonary artery pressure, from 19 to 41 mm Hg. However, during ExE ischemia was abolished (no angina, decrease in mean pulmonary artery pressure to 24 mm Hg, and improvement in MMI) and there was some improvement in left ventricular pump function, whereas pre- and afterload decreased (pulmonary artery pressure by 40%, systemic vascular resistance by 10%), and heart rate, arterial pressure, and myocardial oxygen consumption (MVO2) were all unchanged (p greater than 0.05). Comparative hemodynamics at RE vs RC showed a decrease in pulmonary artery pressure (by 25%) and pulmonary vascular resistance (by 19%) and an increase in heart rate (by 11%), whereas arterial pressure and MVO2 were unchanged (p greater than 0.05). Enoximone did not induce changes in plasma catecholamine, prostaglandin, or thromboxane levels (p greater than 0.05), whereas the atrial natriuretic factor decreased (by 15%), probably because of unloading of the atria during exercise. We concluded that enoximone induces beneficial hemodynamic effects in coronary artery disease without causing ischemia, probably by enhancing myocardial contractility, vasodilation, and improved diastolic properties. PMID- 2563187 TI - Relative efficacy of, and some adverse reactions to, different antihypertensive regimens. AB - Although it is a common belief that all antihypertensive agents are equally effective in reducing blood pressure, there is some evidence to the contrary, both in the general population and when specific patient demographics are considered. In black patients, beta blockers and angiotensin-converting enzyme (ACE) inhibitors have been shown to be less effective at reducing blood pressure than the thiazide diuretics. After age 60, the percentage of responders to beta blockers is less than with calcium antagonists, and a higher percentage of elderly patients also achieve normotensive blood pressure levels with diuretic therapy than with beta blockers. When a thiazide diuretic is added to an ACE inhibitor, beta blocker or calcium antagonist, the number of normotensive responders increases significantly. Combinations of some other agents (i.e., an ACE inhibitor plus a beta blocker) may not, however, improve efficacy. Diuretics, beta blockers and ACE inhibitors are all generally well tolerated, with a 9 to 10% incidence of subjective side effects. The use of calcium antagonists and especially the centrally acting adrenergic inhibitors may result in more frequent adverse effects. Data from long-term, diuretic-based clinical trials do not support the statement that diuretic therapy results in sustained elevations in lipid levels. These trials have shown cholesterol levels to be at or below baseline after long-term diuretic therapy. The use of beta blockers, on the other hand, may result in long-term elevation of triglyceride levels and a slight decrease in high-density lipoprotein cholesterol. Calcium antagonists and ACE inhibitors do not affect lipid levels, and alpha blockers may actually lower cholesterol levels and increase high-density lipoprotein levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563188 TI - Practical issues in drug selection and dosing. AB - Current guidelines recommend initiating antihypertensive therapy at a diastolic blood pressure greater than 90 mm Hg when nonpharmacologic measures have been unsuccessful. The risk of heart attack and stroke is increased regardless of whether the elevation of blood pressure is primarily systolic alone, diastolic alone, or both. In treating mild to moderate hypertension and in prevention and wellness programs in normotensive persons with a family history of the disease, the initial approach should be nonpharmacologic. Patients who remain hypertensive should proceed to drug therapy. Low-dose thiazide diuretics remain the preferred first step in elderly patients. In patients younger than age 40 years, especially those with tachycardia, a beta blocker may be used as the first step. Treatment should be initiated with less than a full dose, only proceeding to a full dose if necessary. If there is any appearance of extrasystoles, or a decrease in potassium levels to below 3.5 mEq/liter in the elderly, a potassium-sparing diuretic combination should be used. Although there is growing evidence that the effects of diuretic drugs on potassium may contribute to arrhythmias and sudden death, there is also increasing data suggesting that conserving electrolytes during diuretic therapy may obviate these ill effects. PMID- 2563189 TI - Review of the long-term controlled trials of usefulness of therapy for systemic hypertension. AB - A number of long-term clinical trials involving over 40,000 patients have been performed to study the effectiveness of antihypertensive therapy in controlling high blood pressure and in reducing the morbidity and mortality associated with hypertension. Only diuretics and beta blockers have been studied in long-term trials to determine their efficacy in reducing cardiovascular morbidity and mortality. The Hypertension Detection and Follow-Up Program (HDFP), Medical Research Council (MRC) trial, European Working Party on Hypertension in the Elderly (EWPHE) trial, Australian Therapeutic Trial in Mild Hypertension, and the Veterans Administration Cooperative Study all showed a reduction in stroke rate. The EWPHE and HDFP trials were the only studies to show a statistically significant reduction in mortality from myocardial infarction. All of these were diuretic-based; in addition, the MRC trial also used a beta blocker as first-step therapy in 1 cohort. The International Primary Prospective Prevention Study in Hypertension and Heart Attack Primary Prevention in Hypertension (HAPPHY) trials compared beta-blocker and non-beta-blocker or diuretic-based therapies and found no significant difference between the treatment groups in the incidence of stroke or cardiac events. Neither study had a control group, so it was impossible to determine if there was any reduction in stroke or cardiac events. The Metoprolol Atherosclerosis Prevention in Hypertension trial, an extension of the HAPPHY trial, showed that smokers receiving the beta blocker metoprolol had a significantly lower cardiovascular mortality rate than those randomized to a diuretic drug. However, subgroup analysis of the HAPPHY data showed no differences in the effect of beta blockers and diuretics in smokers.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563190 TI - Somatostatin versus cimetidine in the treatment of actively bleeding duodenal ulcer: a prospective, randomized, controlled trial. AB - In a double-blind, prospective, randomized trial, the healing effect of somatostatin was compared with that of cimetidine in 67 patients with endoscopically confirmed duodenal ulcer with active bleeding (33 patients were treated with somatostatin, 34 with cimetidine). Intravenous infusion of either drug was administered for 48 or 72 h. Somatostatin had a higher rate of control of bleeding (30 vs. 24 patients at 72 h; p less than 0.05) and had less mean transfusion requirements (4.4 vs. 5.8 units; p less than 0.01). Need for surgical intervention and the mortality figures show a trend in favor of somatostatin. There was no difference in rebleeding rate. PMID- 2563191 TI - Sulphasalazine-induced lupus-like syndrome with cardiac tamponade in a patient with ulcerative colitis. PMID- 2563192 TI - Human population genetic studies of five hypervariable DNA loci. AB - Population genetic studies were performed using DNA probes that recognize five hypervariable loci (D2S44, D14S1, D14S13, D17S79, and DXYS14) in the human genome. DNA from approximately 900 unrelated individuals, subdivided into three ethnic groups (American blacks, Caucasians, and Hispanics) were digested with PstI and were successively hybridized to each DNA probe. The number of distinct DNA fragments identified for each of these regions varies from 30 to more than 80. An allele frequency distribution was determined for each locus and each ethnic group. The results show significant differences, between ethnic groups, in the pattern of distribution as well as in the relative frequency of the most common alleles of D2S44, D14S1, and D14S13 but only small differences in others (i.e., D17S79 and DXYS14). The results presented show that the analysis of these loci can have useful applications in population genetics as well as in identity tests. PMID- 2563193 TI - The clinical and screening age-at-onset distribution for the MEN-2 syndrome. AB - The decision to screen for multiple endocrine neoplasia type 2 (MEN-2) is generally based on family history, the rationale for this approach being the presumed 100% penetrance of the disease. To determine the validity of this presumption we have estimated--by applying modifications of the life-table method -the clinical and screening age-at-diagnosis distributions for MEN-2, using families from the Cancer Research Campaign Medullary Thyroid Cancer Register and one large American family. The clinical penetrance of MEN-2 is shown to be incomplete, an estimated 41% of gene carriers not presenting with symptoms by age 70 on the basis of clinical history. Screening by the standard tests for detecting the earliest manifestations of the syndrome increases the penetrance to an estimated 93% by age 31. There is no evidence of a difference in the age-at diagnosis distributions between maternal and paternal transmission, or among different families, but there is some suggestion of an earlier onset of medullary thyroid cancer in female gene carriers, and of a tendency of pheochromocytoma to cluster in families. These results can be used to calculate risks to relatives of affected individuals, which in turn can be used to guide decisions on which individuals to screen. PMID- 2563194 TI - New polymorphisms at the DXS98 locus and confirmation of its location proximal to FRAXA by in situ hybridization. AB - The locus DXS98, detected with the 1.5-kb anonymous probe p4D-8, was recently shown to be closely linked and proximal to the locus for the fragile X syndrome, with theta = .05 at lod = 3.406, by utilizing a limited number of meioses informative for a two-allele MspI RFLP. Because DXS98 may be the closest available marker to the fragile X locus (FRAXA), we sought to increase its utility for linkage studies by extending its PIC and confirming its localization to Xq27, proximal to FRAXA. We have isolated 15 kb of genomic DNA (lambda 4D8-3) from the DXS98 locus by using p4D-8 to screen a genomic phage library containing partial Sau3A-digested human DNA. Three additional RFLPs for the enzymes BglII and XmnI were found by using the entire lambda 4D8-3 as probe. Combined heterozygosity for the four RFLPs in 25 unrelated females was 48%, as compared with only 28% when the MspI RFLP alone was used. In situ hybridization of unique sequences from lambda 4D8-3 was performed on metaphase chromosomes of lymphocytes and lymphoblasts from patients with the fragile X syndrome. Grains on the X chromosome were significantly clustered at band Xq27. Following fragile site induction, all nine grains in the q27-28 region were proximal to the fragile site. Confirmation of the location of DXS98 proximal to FRAXA and the new RFLPs at this locus make DXS98 more useful for linkage analysis and physical mapping in the region of the fragile X mutation. PMID- 2563195 TI - Assignment of autosomal dominant spinocerebellar ataxia (SCA1) centromeric to the HLA region on the short arm of chromosome 6, using multilocus linkage analysis. AB - A 7-generation kindred with the HLA-linked form of spinocerebellar ataxia (SCA1) was studied to determine whether the SCA1 gene maps centromeric or telomeric to the HLA loci. The DNA markers flanking the HLA-(A-B) region were used for polymorphism studies and multilocus linkage analysis. These two markers are the cDNA for the beta-subunit of HLA-DP, which is centromeric to HLA-(A-B), and the cDNA for coagulation factor XIIIa (F13A), which is telomeric to HLA-(A-B). Haplotypes were constructed using multiple polymorphisms for these two DNA markers, and pairwise linkage analysis revealed a maximum lod score of 2.18 for SCA1 versus HLA-DP at a recombination fraction of .05 and a maximum lod score of 0 for SCA1 versus F13A at a recombination fraction of .50. A possible crossover between HLA-(A-B) and HLA-DP was identified, but lack of samples from key individuals hampered the analysis. To clarify the phase and improve the analysis, the two chromosomes 6 for the crossover individual were separated in somatic cell hybrids. The results strongly favored the probability that the crossover occurred between HLA-(A-B-DR) and HLA-DP with SCA1 segregating with HLA-DP, consistent with a location centromeric to HLA-(A-B). Multilocus linkage analysis was used to evaluate further the location of SCA1 relative to F13A, HLA-(A-B), and HLA-DP; the results indicated that the SCA1 gene locus is centromeric to HLA-DP with odds of 46:1 favoring this most likely location over the second most likely location, i.e., telomeric to HLA-(A-B) between the HLA complex and F13A. PMID- 2563196 TI - Androgen receptor locus on the human X chromosome: regional localization to Xq11 12 and description of a DNA polymorphism. AB - The gene for the androgen receptor, mutations at which cause the X-linked androgen insensitivity syndrome, has been localized to the q11----q12 region of the human X chromosome by analysis, using a cloned cDNA for the androgen receptor, of somatic cell hybrid panels segregating portions of the X chromosome. A moderate-frequency HindIII RFLP has been found which should be useful in genetic linkage analysis of the various inherited forms of androgen insensitivity. PMID- 2563197 TI - Neurotransmitters and peptides modulate the release of immunoreactive corticotropin-releasing factor from cultured human placental cells. AB - The substances stimulating the release of immunoreactive corticotropin-releasing factor from cultured human placental cells were investigated. Monolayer primary cultures of trophoblast cells from pregnant women at term were used. The immunoreactive corticotropin-releasing factor released in the culture medium eluted from high-performance liquid chromatography with the same retention time as human corticotropin-releasing factor. Norepinephrine and acetylcholine increased immunoreactive corticotropin-releasing factor release into the culture medium in a dose-related manner. Epinephrine was partially active, whereas dopamine and serotonin did not induce significant changes of immunoreactive corticotropin-releasing factor release from placental cultures. Angiotensin II, interleukin-1, oxytocin, and arginine-vasopressin also increased placental immunoreactive corticotropin-releasing factor release in a dose-related manner, whereas other peptides (vasoactive intestinal peptide, substance P, somatostatin, atrial natriuretic factor, interleukin-2) were ineffective. These results showed that several neurotransmitters and peptides stimulate the release of immunoreactive corticotropin-releasing factor from placental cells, suggesting their possible involvement in the physiologic regulation of placental immunoreactive corticotropin-releasing factor release during pregnancy and parturition. PMID- 2563198 TI - Inhibitory effect of somatostatin on vagal motoneurons in the rat brain stem in vitro. AB - Effects of somatostatin-14 (SRIF) on membrane electrical properties were studied in rat brain stem slice preparations maintained in vitro. SRIF hyperpolarized the resting membrane potential and decreased the input resistance of more than two thirds of the 85 vagal motoneurons tested in the dorsal motor nucleus of the vagus. These effects persisted under synaptic blockade caused by perfusion with a solution containing tetrodotoxin or a Ca2+-free/high-Mg2+ solution and were dependent on the extracellular SRIF concentration (5 X 10(-8) to 1 X 10(-8) M). The Hill coefficient was estimated to be 2. The reversal potential of SRIF induced hyperpolarization was affected by changing external K+ concentration. The results suggest that, in addition to its well-known peripheral action, SRIF may inhibit secretomotor functions of visceral organs by reducing vagal output in the central nervous system. PMID- 2563200 TI - Evidence for inhibition of opossum LES through intrinsic gastric nerves. AB - We have studied the mechanical responses of the isolated opossum gastroesophageal junction to a variety of stimuli. Lower esophageal sphincter (LES) relaxations were readily produced by gastric stretch and by electrical stimulation of the intramural gastric nerves. Gastric stimulation also led in 60% of stimuli to a contraction of circular muscle (the off response) in the distal esophagus, but in less than 15% of stimuli to a contraction of longitudinal esophageal muscle (the duration response). Mechanical or electrical stimulation of the esophagus led to a mechanical response of the stomach (relaxation, contraction, or both) in 76% of stimuli. The LES relaxation produced by esophageal stimulation was not as long as that produced by gastric stimulation. Direct electrical stimulation of the esophageal or gastric muscle produced a ring contraction that spread readily across the gastroesophageal junction but did not produce a relaxation of the LES or a contraction of the longitudinal esophageal muscle. Thus intramural nervous pathways can coordinate the mechanical activity of the esophagus, LES, and stomach. LES relaxations may occur as part of intrinsic reflexes that mediate gastric accommodation to volume. PMID- 2563199 TI - Glucoregulatory function of thyroid hormones: role of pancreatic hormones. AB - Glucose metabolism was investigated in humans before and 14 days after 300 micrograms L-thyroxine (T4)/day using a sequential clamp protocol during short term somatostatin infusion (500 micrograms/h, 0-6 h) at euglycemia (0-2.5 h), at 165 mg/dl (2.5-6 h), and during insulin infusion (1.0 mU.kg-1.min-1, 4.5-6 h). T4 treatment increased plasma T4 (+96%) and 3,5,3'-triiodothyronine (T3, +50%), energy expenditure (+8%), glucose turnover (+32%), and glucose oxidation (Glucox +87%) but decreased thyroid-stimulating hormone (-96%) and nonoxidative glucose metabolism (Glucnonox, -30%) at unchanged lipid oxidation (Lipox). During somatostatin and euglycemia glucose production (Ra, -67%) and disposal (Rd, -28%) both decreased in euthyroid subjects but remained at -22% and -5%, respectively, after T4 treatment. Glucox (control, -20%; +T4, -25%) fell and Lipox increased (control, +42%; +T4, +45%) in both groups, whereas Glucnonox decreased before ( 36%) but increased after T4 (+57%). During somatostatin infusion and hyperglycemia Rd (control, +144%; +T4, +84%) and Glucnonox (control, +326%; +T4, +233%) increased, whereas Glucox and Lipox remained unchanged. Insulin further increased Rd (+76%), Glucox (+155%), and Glucnonox (+50%) but decreased Ra (-43%) and Lipox (-43%). All these effects were enhanced by T4 (Rd, +38%; Glucox, +45%; Glucnonox, +35%; Ra, +40%; Lipox, +11%). Our data provide evidence that, in humans, T3 stimulates Ra and Rd, which is in part independent of pancreatic hormones. PMID- 2563201 TI - Opioid receptors and the initiation of migrating myoelectric complexes in dogs. AB - The role of endogenous opioids and opioid receptors in the control of migrating myoelectric complexes (MMCs) was studied in conscious dogs implanted with silver silver chloride electrodes. In normal fasted dogs, MMC cycle times were 103 +/- 7 min in the duodenum. During naloxone infusion (1-2 mg/kg iv, then 0.2-1.0 mg.kg 1.h-1 iv) cycle times increased to 219 +/- 29 min (P less than 0.01). Naloxone (2 mg/kg iv, then 1 mg.kg-1.h-1 iv) had no effect on the response of the small intestine to bethanecol (5 mg sc) or to feeding. Pretreatment with naloxone (2 mg/kg iv) 5 min before the administration of motilin (400-500 micrograms/kg iv) did not block the initiation of MMCs by motilin. In separate experiments, animals were pretreated with the positive or negative isomer of the opioid receptor antagonist WIN-44,441 (0.2 mg/kg iv) 5 min before morphine administration. The negative isomer binds to opioid receptors whereas the positive isomer does not. The negative but not the positive isomer antagonized all effects of morphine on intestinal myoelectric activity. These studies suggest that endogenous opioids and opioid receptors may play a role in control of the initiation of MMCs and that motilin and exogenous opioids act via different mechanisms to initiate MMCs. PMID- 2563202 TI - Functional localization of specific receptors mediating gastrointestinal motor correlates of vomiting. AB - The gastrointestinal motor correlates of vomiting consist of two contractile events, 1) a giant retrogradely propagated contraction of the upper small intestine, the retrograde giant contraction (RGC) and 2) a series of post-RGC phasic contractions that occur primarily in the lower small intestine. The effects of cholinergic, dopaminergic, serotonergic, and opioid receptor antagonists and an opioid receptor agonist on vomiting and its gastrointestinal motor correlates initiated by apomorphine (APO), CuSO4, or cholecystokinin octapeptide (CCK-8) were determined in awake dogs. Atropine blocked the retrograde giant contraction only, and hexamethonium blocked all jejunoileal motor responses activated by APO, CuSO4, or CCK-8. Domperidone blocked all effects of APO only, whereas haloperidol, methysergide, 1-(1-naphthyl) piperazine, and fentanyl blocked or inhibited responses to both APO and CuSO4. None of the dopaminergic, serotonergic, or opioid receptor antagonists or the opioid receptor agonist affected the gastrointestinal motor responses to CCK-8. Cinanserin or Sch 23390 had no effect on any of the responses activated by APO, CuSO4, or CCK-8. These results suggested that D2 dopaminergic and 5-HT2 serotonergic receptors of the emetic central pattern generator mediate vomiting and its gastrointestinal motor correlates, whereas opioid receptors may mediate tonic inhibition of these responses. In addition, peripheral muscarinic or nicotinic cholinergic receptors but not peripheral 5-HT2, dopaminergic, or opioid receptors mediate the gastrointestinal motor correlates of vomiting. PMID- 2563203 TI - Ammoniagenesis by cultured human renal cortical epithelial cells: study with 15N. AB - The metabolic fate of 15N-labeled glutamine and glutamate in cultured human renal cortical epithelial cells was investigated. The main goal was to elucidate the major pathways of ammoniagenesis depending on varying H+ concentration. Incubations at pH 7.4 or 6.8 were conducted with either 1 mM [5-15N]glutamine, [2 15N]glutamine, [15N]glutamate, or L-[2-15N]-gamma-glutamylmethylamide. The results demonstrate that acute acidosis had little effect on total ammonia generation from glutamine. However, 15NH3 formation from [5-15N]glutamine was significantly higher at pH 7.4 compared with pH 6.8. Conversely, at pH 6.8, 15NH3 production from either [2-15N]-glutamine or [15N]glutamate was twofold higher than at pH 7.4. Thus the observations indicate that acute acidosis had little effect on net ammonia production from glutamine due to decreased flux through glutaminase and concomitant increased flux through glutamate dehydrogenase. When L-[2-15N]-gamma-glutamylmethylamide was utilized as the sole substrate, significantly higher amounts of 15NH3 and 15N-labeled amino acids were formed at pH 6.8 compared with pH 7.4. Addition of either 1 mM pyruvate or alpha ketoglutarate significantly decreased 15NH3 and increased 15N-amino acid formation from either [2-15N]glutamine or [2-15N]-gamma-glutamylmethylamide. The metabolism of either substrate via transamination reaction was significantly stimulated at acidic pH, presumably due to a depleted pool of alpha-ketoglutarate during the course of the incubations. The data indicate that in addition to glutaminase I and glutamate dehydrogenase, the glutamine aminotransferase (glutaminase II) pathway exists in cultured human renal cells. The data suggest that glutamate dehydrogenase flux and/or the alpha-ketoglutarate dehydrogenase reaction may have an important regulatory role in ammoniagenesis from glutamine and/or glutamate in human kidney during acute acidosis. PMID- 2563204 TI - Regulation of Na+-K+-ATPase activity in kidney proximal tubules: involvement of GTP binding proteins. AB - This study evaluates the involvement of GTP-dependent regulatory proteins (G proteins) in the regulation of Na+-K+-ATPase activity in proximal convoluted tubule (PCT) segments. Single PCT segments were dissected from rat kidney and permeabilized to allow nucleotides and medium free access to the interior of the cell. A GDP analogue that blocks GTP-dependent activation of the G-protein, GDP beta S (400 microM) significantly inhibited PCT Na+-K+-ATPase activity when Na in the medium (Nam) was greater than or equal to 70 mM. The inhibition was attenuated when Nam was 55 and 35 mM and was no longer significant when Nam was 25 mM. GDP beta S had no inhibitory effect on the activity of purified Na+-K+ ATPase. A nonhydrolyzable GTP analogue, GppNHp (50 microM) significantly increased Na+-K+-ATPase activity when Nam was 25 and 35 mM, but not when Nam was 55-140 mM. Dopamine (DA) and DA1 plus DA2 agonists significantly inhibit Na+-K+ ATPase activity. DA inhibition was competitively abolished by GppNHp. In PCT segments from rats pretreated with pertussis toxin, DA and DA1 plus DA2 agonist inhibition of Na+-K+-ATPase activity was abolished. In PCT segments from rats pretreated with cholera toxin, basal Na+-K+-ATPase activity was increased, but DA significantly inhibited Na+-K+-ATPase activity. Na+-K+-ATPase activity in PCT segments is regulated via a G-protein that stimulates Na+-K+-ATPase activity and a DA-activated pertussis toxin-sensitive G-protein that inhibits Na+-K+-ATPase activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563205 TI - Paraventricular stimulation with glutamate elicits bradycardia and pituitary responses. AB - The excitatory neurotransmitter, L-glutamate (0.5 M, pH 7.4), or the organic acid, acetate (0.5 M, pH 7.4), was microinjected (50 nl over 2 min) directly into the paraventricular nuclei (PVN) of pentobarbital sodium-anesthetized rats while arterial blood pressure and heart rate and plasma adrenocorticotropic hormone (ACTH), vasopressin, and oxytocin were measured. Activation of PVN neurons with L glutamate led to increases in plasma ACTH, vasopressin, and oxytocin and a profound bradycardia (approximately 80 beats/min) with little change in arterial blood pressure. Microinjection of acetate had no effect on the above variables. The decrease in heart rate was shown to be dependent on the concentration of glutamate injected and the volume of injectate. The bradycardia was mediated through the autonomic nervous system because ganglionic blockade (pentolinium tartrate) eliminated the response; atropine and propranolol severely attenuated the bradycardia. The bradycardia was greatest when L-glutamate was microinjected into the caudal PVN. Injections into the rostral PVN or into nuclei surrounding the PVN led to small or nonsignificant decreases in heart rate. Focal electric stimulation (2-50 microA) of the PVN also led to decreases in heart rate and arterial blood pressure. These data suggest that activation of PVN neurons leads to the release of ACTH, vasopressin, and oxytocin from the pituitary and a bradycardia that is mediated by the autonomic nervous system. PMID- 2563206 TI - Management of neuroleptic-induced movement disorders: effects of physician training. AB - Residents given a specialized 4-hour course in managing extrapyramidal syndromes prescribed significantly lower neuroleptic doses and were more skilled at diagnosis of extrapyramidal syndromes than residents given standard psychopharmacology training. The results suggest that focused training in extrapyramidal syndromes should be part of residency curricula. PMID- 2563208 TI - Calcium blockers in tardive dyskinesia. PMID- 2563207 TI - "Ecstasy" (MDMA)-induced panic. PMID- 2563209 TI - Comments on posttraumatic stress disorder. PMID- 2563210 TI - Fluphenazine decanoate: a clinical problem? AB - The authors compared the medical records of 20 randomly selected schizophrenic patients who were receiving fluphenazine decanoate with the records of 20 other randomly selected schizophrenic patients who were receiving oral medications only. The fluphenazine decanoate group had a significantly higher mean chlorpromazine equivalent dose (2010 +/- 836) than the oral medication group (373 +/- 384). The two groups did not, however, differ on any of the other variables studied. The findings, in addition to reports in the literature, suggest that the use of fluphenazine decanoate may come at the inadvertent cost of a significantly higher dose of antipsychotic medications in a subgroup of patients. PMID- 2563211 TI - Neuroleptic augmentation with alprazolam: clinical effects and pharmacokinetic correlates. AB - Alprazolam added to stable doses of neuroleptics in nine schizophrenic patients was associated with a 20%-30% mean reduction in positive and negative symptoms, although clinical response was variable and in some patients particularly brisk. The authors examined the possibilities of a pharmacokinetic effect of alprazolam on neuroleptic plasma levels and of a clinical effect of alprazolam. The modest increase in mean neuroleptic plasma levels did not correlate with clinical change, but those patients with the highest alprazolam plasma levels tended to show more robust clinical responses. PMID- 2563212 TI - Delirium with dystonia: a variant of neuroleptic malignant syndrome? PMID- 2563213 TI - Schizophrenia, panic attacks, and antidepressants. PMID- 2563214 TI - A model for determining the influence of hepatic uptake of nondepolarizing muscle relaxants in the pig. AB - Hepatic uptake and distribution of nondepolarizing muscle relaxants in pigs was investigated. A portocaval shunt preparation enabled the determination of the pharmacodynamics of nondepolarizing muscle relaxants both during temporary liver exclusion and intraportal injection in the same animal. To demonstrate the validity of the model in pigs, in a pilot study the influence of hepatic uptake on neuromuscular blockade by pancuronium (n = 3) and its congener Org 6368 (n = 3) was determined. Thereafter, the influence of hepatic uptake on neuromuscular blockade by gallamine (3.4 mg/kg, n = 5), Org 6368 (0.3 mg/kg, n = 5), and vecuronium (0.1 mg/kg, n = 4 and 0.15 mg/kg, n = 5) was studied in pigs anesthetized with pentobarbital. Each pig received one relaxant, which was injected four consecutive times under different conditions. The first injection was into the jugular vein (iv) the second into the portal vein, the third was iv while the liver was excluded for 10 min and the fourth was iv (control). After each injection the onset time, intensity, recovery rate, and total duration of neuromuscular blockade were measured. These variables were compared between the four injections for each relaxant. In the pilot study the influence of hepatic uptake on neuromuscular blockade was similar for pancuronium and Org 6368. For gallamine the onset time, intensity, recovery rate, and duration of action were similar after all four injections. For Org 6368 the variables of neuromuscular blockade were similar after iv and intraportal injection, but exclusion of the liver prolonged the neuromuscular block.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563215 TI - An exaggerated response to beta-adrenergics. AB - Beta-adrenergics are used frequently in the management of asthma. Tremor has been found to be a limiting side effect with the oral or the inhaled forms. We describe one child who developed gross tremors necessitating an extensive neurologic evaluation to eliminate any other cause. With the results of a normal work-up and the reappearance of tremor when challenged again, the diagnosis of increased sensitivity to the tremorogenic effect of beta-adrenergics was made. PMID- 2563216 TI - Review of side effects associated with beta agonists. AB - Beta agonists are a primary drug of choice in the treatment of asthma. Nevertheless, there are significant concerns about side effects in using these medications, especially in certain disease states, in higher dosages, and in severe asthma episodes. Proper patient evaluations and monitoring can reduce significantly these side effects and reduce their health consequences. PMID- 2563217 TI - Effects of somatostatin and oral potassium administration on terbutaline-induced hypokalemia. AB - Terbutaline, a beta 2-adrenergic agonist, has been shown to cause hypokalemia and an increase of plasma glucose and serum insulin concentrations. We considered that terbutaline-induced hypokalemia may be due to the insulin-induced shift of potassium (K+) from the extracellular to the intracellular space. If so, then inhibition of insulin secretion by somatostatin would prevent terbutaline-induced hypokalemia. Further, we wondered whether oral potassium pretreatment could prevent terbutaline-induced hypokalemia. Therefore, 10 healthy volunteers (5 men, 5 women; mean age, 23 yr +/- 3 SD) received either sodium chloride (NaCl) or somatostatin intravenously together with 0.25 mg terbutaline subcutaneously in a double-blind crossover design. On a third test day, they received 39 mval of K+ powder orally before terbutaline injection in an open trial. Terbutaline caused a significant decrease of K+ (from 3.96 +/- 0.08 to 3.3 +/- 0.13 mmol/L +/- SEM; p less than 0.0005), accompanied by a significant increase in plasma glucose (from 83 +/- 3.6 to 101 +/- 4.4 mg/dl +/- SEM; p less than 0.01) and serum insulin concentrations (from 11.7 +/- 0.9 to 19.9 +/- 1.1 microU/ml +/- SEM; p less than 0.001), confirming earlier data. Somatostatin pretreatment inhibited the terbutaline-induced hypokalemia; the small fall of K+ (from 3.7 +/- 0.08 to 3.5 +/- 0.2 mmol/L) was no longer significant. Insulin secretion was completely blocked by somatostatin, leading to an even more pronounced increase of blood glucose. Hypokalemia after terbutaline injection was not prevented by oral potassium pretreatment. In summary, the present findings confirm that terbutaline induced hypokalemia is associated with increased plasma glucose and insulin levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563218 TI - [Anti-tumor natural products isolated from marine organisms]. AB - Marine organisms comprise over half a million species. Due to their unusual living environment as compared with terrestrial organisms, marine organisms produce a variety of metabolic substances which often have various unprecedented chemical structures. In recent years, an increasing number of marine natural products have been reported to exhibit various biological activities such as antimicrobial, physiological, and pharmacological ones. Some metabolites have also been noted by their significant toxicities. The effort to find out anti tumor substances from marine organisms has been also undertaken in recent years, and several novel compounds (e.g. peptide, polyether, alkaloid, prostanoid etc.), with anti-tumor activities, have been isolated from marine sponges, octocorals, marine algae, tunicates, nudibranchs, bryozoans, and so on. In this article, those chemically clarified anti-tumor compounds isolated from marine organisms were reviewed. PMID- 2563219 TI - Pediatric implications of heterozygous familial hypercholesterolemia. Screening and dietary treatment. AB - Heterozygous familial hypercholesterolemia (FH) is completely expressed at birth and in childhood by significant elevations of plasma total and low density lipoprotein (LDL) cholesterol levels. High density lipoprotein cholesterol levels can be low in FH. Screening of children for FH using a LDL cholesterol level is efficient in families with known FH, while for general population screening, the LDL cholesterol level is too nonspecific. Newer cellular and molecular biologic approaches of screening for FH promise to be more specific. A diet low in cholesterol (less than 200 mg/day), total fat (30% of calories), and saturated fat (less than 10% of calories) but moderately enriched in polyunsaturated fat (up to 10% of calories) will lower the total and LDL cholesterol levels about 10% to 15% in most heterozygous FH children. Many children will eventually require the addition of a drug to achieve satisfactory lowering of the LDL cholesterol level. The early detection and treatment of FH offers the optimal approach to the prevention of premature coronary artery disease. PMID- 2563220 TI - Genetic aspects of familial hypercholesterolemia and its diagnosis. AB - Heterozygote familial hypercholesterolemia (FH) is a frequent genetic trait (one in 500) that predisposes to coronary atherosclerosis. Male heterozygotes often develop clinical manifestations of coronary heart disease in their fourth or fifth decade, females, about 10 years later. Other risk factors for ischemic heart disease like smoking interact with the gene for FH. Homozygotes and compound heterozygotes (i.e., those who carry two different FH genes) are very rare (one in 1,000,000) have severe hypercholesterolemia with xanthomas, and develop coronary heart disease early in life. FH is caused by one of many different mutations (deletions, insertions, missense, or nonsense mutations) affecting the well-defined low density lipoprotein (LDL) receptor gene on chromosome 19. Some populations that started with a small founder group like the Afrikaners in South Africa and the French Canadians carry only a few FH mutations. The diagnosis of heterozygote FH is often difficult, since physical findings (xanthomas) are frequently absent, and overlap of laboratory test values between affected and nonaffected subjects occurs. The a priori probability of heterozygosity of FH varies from one in 500 in population studies to one in two in family studies and must be considered when assessing borderline quantitative test results based on cholesterol, LDL cholesterol, or LDL-receptor assays. The mutational heterogeneity of FH makes it very difficult to use DNA methodology for population detection. However, direct molecular diagnosis by appropriate DNA probes is possible if the specific LDL receptor defect is known. Indirect molecular diagnosis based on genetic linkage of a common DNA variant of the LDL receptor to the basic defect is often feasible but requires family studies. PMID- 2563221 TI - Gene probes in diagnosis of familial hypercholesterolemia. AB - In the United Kingdom, about 5% of patients with familial hypercholesterolemia (FH) have a detectable deletion or rearrangement of part of the LDL-receptor gene, which results in the detection of shorter or abnormally sized fragments of the LDL-receptor gene in a Southern blot hybridization. This gene deletion can be used for following the inheritance of the defective gene and for diagnosis in the families of these individuals. In the families of the remaining patients, diagnosis may be possible using linked restriction fragment length polymorphisms (RFLPs) detected with the LDL-receptor probe. There are now 10 common RFLPs of the LDL-receptor gene, with variable sites in the 3' half of the gene. Over 80% of patients are heterozygous for at least one of these RFLPs, and, therefore, RFLPs are potentially informative for DNA diagnosis. For a fetus at risk of homozygous FH, antenatal diagnosis may also be possible using these methods. However, family studies require samples to be available from affected or unaffected relatives of the patient, and this limits the applicability of the tests. For some mutations, the base-pair change causing the defect in the LDL receptor itself creates or destroys a site for a restriction enzyme. Such "mutation-specific" RFLPs could be used for population screening, but, so far, such use has only been reported for the FH mutation common in Lebanon. In the future, it may be possible to develop mutation-specific oligonucleotide probes for the diagnosis of FH. These would be appropriate for screening populations or patients with hyperlipidemia. This information may also be useful if different mutations require different therapeutic strategies. PMID- 2563222 TI - Changes in mitochondrial activity caused by ammonium salts and the protective effect of carnitine. AB - Ammonium salts added to isolated rat liver mitochondria deviate alpha ketoglutarate to glutamate synthesis, thus decreasing its availability as respiratory substrate. As a consequence a decrease of respiratory rate is observed which is paralleled by progressive mitochondrial swelling. It was demonstrated that L-carnitine may abolish this swelling thus improving structural and metabolic state of mitochondria. PMID- 2563223 TI - Alterations in c-abl gene methylation in cells transformed by phagocyte-generated oxidants. AB - DNA from 10T1/2 cells transformed by activated neutrophils was analyzed for restriction length polymorphisms (RFLPs) in cellular homologues of retroviral oncogenes, and consistent RFLPs were found in MspI sites of the c-abl gene of all PMN-transformed cell lines. MspI digests probed with c-myc, v-Ki-ras, v-Ha-ras or v-mos showed no RFLPs, and none were observed in EcoRI, PstI, HindIII, BamHI, SmaI, Sau3a, MboI, HhaI, or TaqI digests probed with v-abl. Analysis of HpaII digests supports the conclusion that c-abl RFLPs result from differential methylation of the CCGG HpaII/MspI recognition sequence. MspI RFLPs in the c-abl gene may provide markers for oxidant-related genetic injury. PMID- 2563224 TI - Metabolic responsiveness to phorbol ester and activity of protein kinase C in isolated hepatocytes from partially hepatectomized rats. AB - The ability of isolated rat hepatocytes to respond to phorbol-12-myristate-13 acetate (PMA) with acute stimulation of de novo fatty acid synthesis was markedly depressed at 4, 22 and 48 h after partial hepatectomy (PH). This desensitization was not due to surgical stress as shown by comparison with hepatocytes from sham operated animals. Moreover, the total activity of protein kinase C (PK-C), the principal phorbol ester receptor, was not down-regulated at 22 h after partial hepatectomy. Partial hepatectomy rather caused a small but distinct shift in subcellular PK-C distribution toward the particulate fraction thereby suggesting a modest activation of PK-C. We conclude that the PH-induced desensitization to PMA occurs at a point beyond PK-C activation. PMID- 2563225 TI - Amino acid residues essential for catalysis by peptidyl dipeptidase-4 from Pseudomonas maltophilia. AB - To assess residues essential for catalysis by prokaryotic peptidyl dipeptidase-4, the enzyme was subjected to chemical modification by a series of reagents. Treatment with either tetranitromethane or N-acetylimidazole abolished catalytic activity. Hydroxylamine reversed inactivation by acetylimidazole only. Thus, an essential tyrosine is indicated. Enzymatic activity also was quenched by either trinitrobenzenesulfonic acid or diethyl pyrocarbonate. Inactivation by these reagents was not reversed by hydroxylamine. These data suggest an essential lysine. The competitive inhibitor Phe-Arg protected partially against inactivation by tetranitromethane, and fully against inactivation by N acetylimidazole. The substrate Hip-Phe-Arg protected against inactivation by trinitrobenzenesulfonic acid and diethyl pyrocarbonate. Thus, both tyrosine and lysine are located at the catalytic site. PMID- 2563226 TI - Infectious complications of cyclophosphamide treatment for vasculitis. AB - Fifteen patients with Wegener's granulomatosis, polyarteritis nodosa, or isolated angiitis of the central nervous system were treated with cyclophosphamide according to a widely used regimen. Seventeen clinical episodes of infection were observed over 201 patient-months of cyclophosphamide therapy, and 2 patients died of pneumonia. Notably, neither the incidence of leukopenia nor the dosage or duration of cyclophosphamide or corticosteroid therapy correlated well with infection, which occurred most frequently in men over 60 years of age. Patients with Wegener's granulomatosis appeared to be at greater risk of infection than those with the other forms of vasculitis. These results suggest that this treatment regimen may not be as safe as was previously thought. PMID- 2563227 TI - Superoxide dismutase deficiency and the toxicity of the products of autooxidation of polyunsaturated fatty acids in yeast. AB - Mutants of Saccharomyces cerevisiae, deficient in cytosolic superoxide dismutase and catalase activities were used to study the role of various oxygen species in the process of lipid peroxidation in yeast cells. Lipid peroxidation does not occur normally in yeast, because this organism is unable to form fatty acids with more than one double bond, whereas under physiological conditions, only fatty acids with at least two double bonds undergo this process. The fatty acid content of cellular lipids was modified by growing the cells in anoxia in the presence of oleic or linolenic acid. Toxic effects of oxygen were observed almost exclusively in those cells of yeast mutants deficient in superoxide dismutase, which contain linolenic acid in cellular lipids. Hypersensitivity of the mutant cells, however, results mainly from toxic effects of the products of autooxidation of extracellular fatty acids. These facts suggest that superoxide dismutases are in some way involved in preventing toxic effects of the products of lipid peroxidation and to some extent prevent the process of lipid peroxidation. PMID- 2563228 TI - Regulation of hepatic parenchymal and non-parenchymal cell function by the diadenine nucleotides Ap3A and Ap4A. AB - The diadenine nucleotides diadenosine 5',5"-P1,P3-triphosphate (Ap3A) and diadenosine 5',5"-P1,P4-tetraphosphate (Ap4A) can be released from platelets and were shown to act as long-lived signal molecules. Accordingly, we studied their potential effect on hepatic metabolism. In isolated perfused rat liver, Ap3A and Ap4A increase the portal pressure, lead to a transient net release of Ca2+, complex net K+ movement across the liver plasma membrane and stimulate hepatic glucose output and 14CO2 production from [1-14C]glutamate. These responses resemble that obtained with extracellular ATP. This and studies on the additivity of ATP and Ap4A effects suggest similar mechanisms mediating the ATP and diadenine nucleotide effects in the liver. Ap3A and Ap4A increased the activity of glycogen phosphorylase a in isolated hepatocyte suspensions by about 100%, pointing to a direct effect of these nucleotides on hepatic parenchymal cells. A response of hepatic non-parenchymal cells to diadenine nucleotide infusion is suggested by a marked stimulation of thromboxane and prostaglandin D2 release from perfused liver. Studies with the thromboxane A2 receptor antagonist BM 13.177 (20 microM) show that the pressure and glucose response to the diadenine nucleotides is partially mediated by this thromboxane formation. Studies with retrograde and sequential liver perfusions suggest a less efficient degradation of the diadenine nucleotides during a single liver passage compared to extracellular ATP. The data suggest that Ap3A and Ap4A are potential regulators of hepatic hemodynamics and metabolism, involving complex interactions between hepatic parenchymal cells and hepatic non-parenchymal cells, including eicosanoids as signal molecules. PMID- 2563229 TI - Plasma HVA, neuroleptics, and dopaminergic plasticity. PMID- 2563230 TI - A note on the drug-induced arterialization of venous blood in schizophrenics: possible microcirculation disturbances. PMID- 2563231 TI - Increased saccadic distractibility in tardive dyskinesia: functional evidence for subcortical GABA dysfunction. AB - In mammals, GABAergic projections from the substantia nigra reticulata to the superior colliculus provide tonic inhibition to tectal neurons involved in the generation of saccades. Dysfunction of this pathway has been shown to produce saccadic "distractibility" in the experimental monkey. In two oculomotor paradigms, control of saccadic eye movements was tested in chronic schizophrenic patients with (n = 18) and without (n = 16) tardive dyskinesia (TD) and normal controls (n = 8). The three groups were matched by mean age; the TD and non-TD patient groups had similar duration of illness, benztropine and chlorpromazine equivalent doses and educational levels. A twofold increase in saccadic distractibility was observed in TD compared to non-TD schizophrenic patients, and both patient groups demonstrated a greater saccadic distractibility than normals. Furthermore, schizophrenic patients (both with and without TD) showed significantly increased latency for "volitional" saccades compared to the normal controls. These findings may provide further evidence for basal ganglia GABA dysfunction in tardive dyskinesia, as well as demonstrate oculomotor abnormalities in schizophrenic individuals. PMID- 2563232 TI - EEG coherence in unmedicated schizophrenic patients. AB - We have recently shown that electroencephalogram (EEG) coherence data recorded with common reference methods, including those obtained from schizophrenics, are confounded by power and phase effects. Three published reports using bipolar recordings found that EEG coherence was higher in schizophrenics; however, only medicated patients were studied. To extend these findings, we measured EEG coherence from bipolar EEG recordings in unmedicated schizophrenics (n = 10), affective disorder patients (n = 8), and normal controls (n = 13) during resting and task conditions. Seven schizophrenics were restudied after a period of neuroleptic treatment. Schizophrenics had higher across-task interhemispheric (p less than 0.05) and intrahemispheric (p less than 0.04) coherence in the theta band and tended to have higher intrahemispheric alpha coherence (p less than 0.08). Medication treatment was associated with clinical improvement and increases in spectral power, but not with changes in coherence values. These results confirm those obtained by earlier investigations and suggest that increased coherence reflects the presence of anomalous cortical organization in schizophrenics rather than medication effects or transient states related to acute clinical disturbance. PMID- 2563233 TI - Alterations in sleep polygraphy after neuroleptic withdrawal: a putative supersensitive dopaminergic mechanism. AB - Although a number of studies have reported sleep disturbances following neuroleptic withdrawal, a full description of such changes in sleep architecture is not available. Polysomnographic, plasma prolactin, and clinical measurements were carried out in a small number of patients on chronic neuroleptic treatment and after drug withdrawal. Preliminary findings show that in these chronically treated schizophrenic patients with and without tardive dyskinesia (TD), abrupt neuroleptic withdrawal induces reductions in total sleep, rapid eye movement (REM) sleep, and plasma prolactin. Furthermore, an increase in delta sleep was observed only in patients without TD. The REM suppression occurred significantly earlier in TD patients compared to the non-TD schizophrenic patients. These changes were transient, and both sleep measures and plasma prolactin stabilized during the 2-4 weeks after withdrawal to levels somewhere between the values observed during chronic treatment and withdrawal (week 1) periods. As the withdrawal-induced exaggerated changes mimicked the dopamine agonist effect on these sleep and hormonal measures, one can hypothesize that the observed changes are due to unmasking of supersensitive dopamine receptors following drug cessation. Normalization of these receptors and/or adaptational changes in other nondopaminergic system(s) can hypothetically explain the eventual stabilization of these measures during the following weeks. PMID- 2563234 TI - Imbalance of deoxyribonucleoside triphosphates, DNA double-strand breaks, and cell death caused by 2-chlorodeoxyadenosine in mouse FM3A cells. AB - The mechanism of the cytotoxic action of 2-chlorodeoxyadenosine in mouse FM3A cells was investigated. Imbalance of the dNTP pools occurred within 3 h of treatment with 20 microM 2-chlorodeoxyadenosine; the dATP and dGTP pools were depleted and the dTTP pool increased. 2-Chlorodeoxyadenosine added to the culture medium broke mature DNA strands, giving fragments of 100-200 kilobase pairs as found by orthogonal-field-alternation gel electrophoresis. DNA strand breaks, measured by this technique, were observed in the treated cells about 12 h after the addition. The cells also lost viability at about 12 h. Breaks in the single and double strands of DNA, as measured by alkaline and neutral filter elution, became evident 18 h after treatment with 20 microM 2-chlorodeoxyadenosine; there were as many single-strand breaks as would be caused by 130 rads of gamma-ray irradiation. Double-strand breaks were equivalent to those caused by 2180 rads of gamma-ray irradiation. Comparison of the ratio of single- and double-strand breaks caused by 2-chlorodeoxyadenosine to that following radiation suggested that 2-chlorodeoxyadenosine broke only double strands. Cycloheximide inhibited the breakage of DNA double strands and the cell death caused by this compound. Flow cytometric studies of cytostasis brought about by 2-chlorodeoxyadenosine in FM3A cells showed that cells accumulated in the earlier part of the S phase. 2 Chlorodeoxyadenosine decreased DNA synthesis more than RNA or protein synthesis. The breaks in double strands of DNA were probably important in the cell death caused by 2-chlorodeoxyadenosine. The intracellular dNTP imbalance may trigger these events. PMID- 2563235 TI - Effects of respiration and vasodilation on venous volume in animals and man, as measured with an impedance catheter. AB - Venous return determines cardiac preload and is in turn affected by respiration and vasodilation. The purpose of the present study was to examine the dynamics of venous return and venous volume, using impedance volume measurements in the venous system. In order to develop a methodology for the assessment of venous volume and venous return in man, we first studied 17 endotracheally intubated and ventilated anesthetized closed-chest dogs. We measured central venous and inferior vena cava (IVC) pressure (micromanometer) and volume (impedance catheter). Studies were done above and below the diaphragm with normal ventilation, with positive end-expiratory pressure (PEEP), and with beta adrenergic blockade and i.v. nitroglycerin. Intrathoracic IVC volume fell and extrathoracic IVC volume rose with lung inflation, while PEEP raised extrathoracic IVC volume and lowered intrathoracic IVC volume. Nitroglycerin lowered intrathoracic IVC volume. Beta blockade did not affect IVC volume, ventilatory variation, or response to PEEP and nitroglycerin. We performed similar studies in 14 human subjects during normal quiet respiration, with measurements above and below the diaphragm, and with interventions including Valsalva maneuver and i.v. nitroglycerin. Intrathoracic IVC volume fell and extrathoracic IVC volume rose with expiration and Valsalva maneuver. Nitroglycerin again lowered intrathoracic IVC volume. We conclude that venous volume and the dynamics of venous return can be assessed in animals and man with an impedance catheter. Specifically, we show the divergent effects of respiration, ventilation, PEEP, and nitroglycerin on IVC volumes above and below the diaphragm. Beta-adrenergic blockage does not appear to play a role in altering any of these effects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563236 TI - Ploidy and karyotype of hepatocytes isolated from enzyme-altered foci in two different protocols of multistage hepatocarcinogenesis in the rat. AB - The ploidy and karyotypes of hepatocytes isolated from the livers of rats subjected to the protocols of Peraino et al. and of Solt and Farber were determined by the examination of such cells in primary culture. A study of 100 or more metaphases from each of five rats on each protocol revealed that 75-80% of gamma-glutamyltranspeptidase-positive (GGT+) hepatocytes isolated from livers of rats in either protocol were diploid, whereas only 23-33% of GGT- cells were diploid. Fifty percent or more of the karyotypes of hepatocytes from livers of rats receiving the Solt-Farber protocol exhibited one or more chromosomal breaks, whereas hepatocytes from livers of rats subjected to the Peraino protocol showed no increase in chromosomal breakage over that in normal controls. These studies demonstrate that the majority of GGT+ cells from altered hepatic foci are diploid and that the greater toxicity of the Solt-Farber protocol over that of Peraino is correlated with marked chromosomal breakage of GGT+ hepatocytes. PMID- 2563237 TI - Anti-inflammatory actions of enprofylline, a modified xanthine, in the canine forelimb. AB - It has been previously reported that enprofylline (3-propyl xanthine) prevents histamine-mediated edema formation in the guinea pig lung. To further assess the potential anti-inflammatory effects of enprofylline, we infused it intra arterially into the canine forelimb before and during a local intra-arterial infusion of histamine (4 micrograms/min) while monitoring forelimb skin lymph parameters. Infusion of enprofylline at 2 mg/min significantly decreased forelimb arterial pressures and increased heart rate and pulse pressure. Subsequent infusion of histamine caused a further reduction in forelimb arterial pressures and an increase in lymph flow, protein concentration, and protein transport similar to that seen with the infusion of histamine alone. Infusion of enprofylline at 5 mg/min decreased forelimb arterial pressures and systemic pressure. Subsequent histamine infusion further reduced forelimb arterial pressures, but the increase in lymph parameters was markedly attenuated. Enprofylline infused at 10 mg/min also decreased forelimb arterial and systemic pressures, but subsequent histamine infusion was essentially without effect on lymph parameters. To assess the role of catecholamines in enprofylline-mediated attenuation of histamine edema formation, we infused enprofylline at 5 mg/min in the presence of a beta 2-receptor blockade produced by the intra-arterial infusion of ICI 118551. The effects of enprofylline and histamine on vascular pressures were similar to those seen in the absence of beta 2-receptor blockade, but lymph flow, protein concentration, and protein transport increased similar to that seen with histamine alone. These data indicate that enprofylline is capable of attenuating histamine-induced increases in microvascular permeability, but this action of enprofylline is of an indirect nature, mediated through the release of catecholamines. PMID- 2563238 TI - Heterogeneous microvascular coronary alpha-adrenergic vasoconstriction. AB - We tested the hypothesis that humoral or neurogenic alpha-adrenergic activation in the coronary circulation would produce heterogeneous vascular reactions. To accomplish this, the epicardial coronary microcirculation was viewed through an intravital microscope using stroboscopic epi-illumination. Microvascular diameters were measured under control conditions during beta-adrenergic blockade (propranolol 1 mg/kg) and beta-adrenergic blockade with pacing; during coronary alpha-adrenergic activation in the presence of beta-adrenergic blockade with three doses of norepinephrine infusion (0.1, 0.5, and 1.0-2.0 micrograms/kg/min) or three frequencies of bilateral stellate nerve stimulation (2, 10, and 20 Hz); and during combined alpha- and beta-adrenergic blockade (phentolamine 2 mg/kg and propranolol 1 mg/kg). Diameters of both arterial and venous vessels were reduced during beta-adrenergic blockade but returned back to baseline with pacing. At the lowest level of norepinephrine infusion or frequency of bilateral stellate stimulation, microvessel constriction was not observed. At the higher doses of norepinephrine a -5.1 +/- 0.9% (1.0-2.0 micrograms/kg/min) and a -4.0 +/- 1.1% (0.5 micrograms/kg/min) decrease in diameter of arterial vessels greater than 100 microns in diameter were observed (p less than 0.05). At 10 Hz and 20 Hz of stellate stimulation, diameter decreased by -4.8 +/- 1.9% and -4.4 +/- 2.1%, respectively, in these relatively large vessels. Small coronary arterioles (less than 100 microns diameter) dilated significantly during the highest levels of nerve stimulation (9.2 +/- 2.5% increase in diameter) or infusion rate of norepinephrine (13.6 +/- 2.7% increase in diameter) (p less than 0.05). These constrictor and dilator responses were abolished following combined alpha- and beta-adrenergic blockade. Norepinephrine infusion resulted in a decrease in diameter of coronary veins and venules (7.2 +/- 1.3%) (p less than 0.05), whereas stellate stimulation did not significantly reduce venous and venular diameters. In summary, the coronary venous and venular vasculature responds to alpha adrenergic activation from circulating norepinephrine but is not affected by stellate stimulation. In contrast, stellate stimulation and norepinephrine infusion elicit similar responses in the coronary arterial and arteriolar microvasculature. Constriction occurs in vessels greater than 100 microns in diameter, whereas dilation predominates in vessels less than 100 microns in diameter. Such heterogeneous arterial responses would undoubtedly result in a redistribution of coronary vascular resistance toward larger coronary arteries and arterioles. PMID- 2563239 TI - Exercise time: a possible source of misleading results during long-term pharmacological studies by multiple stress testings in coronary artery disease. AB - The assessment of chronic pharmacological treatment of stable angina requires serial exercise stress testings. It is well known that exercise tolerance can be improved by the training effect of performing repeated testings. Our study investigated the values of heart rate, systolic blood pressure, rate-pressure product, and duration of exercise at 0.1 mV ST depression during exercise and the same parameters plus the maximal ST-segment depression at peak exercise, collected from three different tests. The first and second were performed at one week intervals before, and the third (75 days after the first), was performed after a double-blind study with a drug versus placebo. We found a significant increase of exercise duration at 0.1 mV ST depression and at peak exercise, while 6 of 12 patients increased exercise duration from the second to the third test. Individual variability of exercise duration showed increasing values, ranging from 0 to 71% (first vs. third test). In contrast, the ratio of heart rate and systolic blood pressure did not differ between the tests. Our data criticized the use of mean values of exercise time for pharmacological studies; moreover, individual variability could affect results independently of drug or placebo administration. These findings should be taken into account in order to exclude misleading results. PMID- 2563240 TI - Direct inhibitory effects of a somatostatin analog, SMS 201-995, on AR4-2J cell proliferation via pertussis toxin-sensitive guanosine triphosphate-binding protein-independent mechanism. AB - Somatostatin has been demonstrated to negatively regulate pancreatic growth in vivo. In this study we used the AR4-2J rat pancreatic acinar tumor cell line to investigate the effect of a stable somatostatin analog, SMS 201-995 (SMS) on cell proliferation. SMS induced an antiproliferative effect on both serum or epidermal growth factor (EGF)-induced cell proliferation; exposure of the cells for 48 h to SMS caused a slight inhibition of serum-induced proliferation (maximal inhibition, 26%) and abolished the growth-promoting effect of EGF. Maximal effect was observed with 10 nM SMS, and half-maximal (IC50) effect with 0.06-0.1 nM SMS. Binding studies with an iodinated derivative of SMS, [125I-Tyr3]SMS, revealed the presence of a single class of high affinity binding sites on AR4-2J plasma membranes with an equilibrium dissociation constant of 0.2 +/- 0.03 nM and a binding site number of 1.1 +/- 0.07 pmol/mg protein. Addition of the nonhydrolyzable GTP analog, guanosine 5-[gamma-thio] triphosphate (GTP gamma S), increased the rate of dissociation of the specifically bound peptide in agreement with the coupling of somatostatin receptors with a GTP-binding regulatory protein. The good agreement between the IC50 for SMS inhibition of cell proliferation and the apparent Kd for binding indicates that the characterized binding sites are the somatostatin receptors that mediate the antiproliferative effect of SMS. When cells were grown in serum-free medium EGF stimulated AR4-2J cell proliferation with half-maximal (ED50) and maximal effects at 0.6 and 10 nM EGF, respectively. This stimulatory effect of EGF was mediated by specific receptors, since binding studies with [125I]EGF indicated that AR4-2J cells contained a single class of EGF receptors (13,000 sites/cell), with an affinity constant for [125I]EGF (Kd = 0.9 +/- 0.09 nM) close to the ED50 for EGF stimulation of cell growth. To examine if SMS-induced growth inhibition involved a cAMP-dependent mechanism we first studied the effect of SMS on cAMP production. SMS had no effect on basal cAMP, but completely inhibited VIP-stimulated cAMP production with an IC50 of 0.2 nM. Pertussis toxin, which is known to abolish the inhibitory effect of somatostatin on adenylate cyclase activity in AR4-2J cells, did not reverse the ability of SMS to inhibit cell proliferation as well as EGF induced cell proliferation. These data indicate that the antiproliferative effect of SMS does not involve the GTP-binding protein-mediated negative coupling of somatostatin receptors to adenylate cyclase.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2563241 TI - A tyrosine aminotransferase glucocorticoid response element also mediates androgen enhancement of gene expression. AB - A glucocorticoid response element of the tyrosine aminotransferase gene was demonstrated to mediate effects of androgen when located upstream of a heterologous promoter linked to the chloramphenicol acetyl transferase gene (CAT). PMID- 2563242 TI - Glucopenia-mediated release of somatostatin from incubated rat hypothalamus: monosaccharide specificity and role of glycolytic intermediates. AB - Rat hypothalamic somatostatin (SRIF) release in vitro is inversely related to medium glucose concentration and is stimulated by agents interfering with neural glucose uptake or metabolism. The present studies examined monosaccharide specificity of this effect by comparing the ability of glucose and nonmetabolized sugars (ribose, fructose, and galactose) to reverse glucopenia-mediated SRIF release. Removal of glucose from incubation medium resulted in a 7-fold stimulation of hypothalamic SRIF release (24.9 +/- 3.0 to 169.0 +/- 46.2 pg/5 min) that was promptly and fully reversed by readdition of glucose (21.5 +/- 6.0 pg/5 min). Depolarizing concentrations of potassium (40 mM) added to medium after incubation in the presence or absence of glucose produced similar 4-fold biphasic stimulation of SRIF release, suggesting that tissue viability was unimpaired by short term glucopenia. By contrast, nonmetabolized sugars (ribose, fructose, or galactose) at concentrations up to 28 mM were unable to reverse glucopenia mediated hypothalamic SRIF release. These findings suggest that neural metabolism of glucose specifically influences hypothalamic SRIF release, an effect likely to depend on glycolysis with resulting energy production. This explanation is supported by the finding that the glycolytic intermediates dihydroxyacetone and glyceraldehyde were partly, and pyruvate completely, able to restore glucopenia mediated SRIF release toward basal, with a maximal effect at 14 mM. Our present studies suggest that rat hypothalamic SRIF release reflects ambient glucose status, is stimulated by glucopenia, and may explain inhibition of GH secretion in the rat during hypoglycemia. PMID- 2563243 TI - Covalent labeling of the somatostatin receptor in rat anterior pituitary membranes. AB - The molecular characteristics of the somatostatin (SRIF) receptor were investigated by covalently cross-linking [125I-Tyr11]SRIF to rat anterior pituitary membranes using three heterobifunctional cross-linking agents, N-5 azido-2-nitrobenzoyloxysuccinimide, N-hydroxysuccinimidyl-4-azidobenzoate, and N succinimidyl-6-(4'-azido-2'-nitrophenylamino) hexanoate, and the homobifunctional agent disuccinimidyl suberate. Sodium dodecyl sulfate-gel electrophoresis followed by autoradiography revealed two SRIF-binding proteins with apparent mol wt (Mr) of 69,000 and 66,000 that were selectively labeled by the four cross linking agents. When cross-linking was performed with N-5-azido-2 nitrobenzoyloxysuccinimide, both proteins migrated as a broad band centered at 68,000; however, with N-hydroxysuccinimidyl-4-azidobenzoate, the band was resolved into 69,000 and 66,000 Mr components. N-Succinimidyl-6-(4'-azido-2' nitrophenylamino) hexanoate covalently labeled the 69,000 Mr protein and a minor species with a Mr of 45,000-47,000. Cross-linking with disuccinimidyl suberate labeled only the 66,000 Mr band. Labeling of both bands was specific, since affinity labeling with each of the four agents was abolished when 1 microM cyclic SRIF was included in the binding reaction. Binding of [125I-Tyr11]SRIF to membranes and labeling of the 69,000 and 66,000 Mr SRIF-binding species were similarly inhibited in a dose-dependent manner by unlabeled SRIF. Radiolabeling of both proteins was specifically displaced by 1 microM SRIF-28 and [D-Trp8,D Cys14]SRIF, but not by oxytocin. Moreover, the extent of radiolabel incorporation into both components was dependent on the concentration of [125I-Tyr11]SRIF in the binding reaction. These results demonstrate the presence of two SRIF-binding proteins in rat anterior pituitary membranes that show characteristics of the SRIF receptor. PMID- 2563244 TI - A critical evaluation of cysteamine as a tool to deplete somatostatin in the rat central nervous system. AB - The wide central nervous system (CNS) distribution of somatostatin (SRIF) as well as the well documented reduction in SRIF concentration in the cerebral cortex in patients with Alzheimer's disease have served as an impetus for studies of this peptide's neurobiological role in the brain. These studies were designed to evaluate the efficacy of centrally administered cysteamine (CYS) as a tool to deplete SRIF in the hypothalamus (HYP) and extrahypothalamic brain areas. Somatostatin was measured by RIA in the frontal cortex (COR), hippocampus (HIP), and HYP in rats after seven daily infusions of CYS into unilateral cannulae stereotaxically positioned into either the lateral ventricle (LV; 300 micrograms/2 microliters) or the dorsal HIP (100 micrograms/2 microliters), and after single (300 mg/kg) or daily (100 mg/kg) sc injections; rats were killed 4 or 24 h after the last injection. After LV infusions, the SRIF concentration was significantly reduced only in the HYP (35% at 4 h and 27% at 24 h). After HIP infusions, the SRIF concentration was significantly reduced only in the HYP at 4 h (23%); no reductions were observed at 24 h. Both a single and repeated sc administrations of CYS reduced SRIF in the HYP only 24 h after treatment (54% and 50%, respectively). Acute sc CYS reduced SRIF in the COR (23%) and the HYP (29%) 4 h after treatment; repeated sc CYS reduced SRIF in the COR (25%) and the HYP (63%). Although the reduction of SRIF in the HYP was increased by repeated sc dosing, the reduction of extrahypothalamic SRIF by sc CYS was relatively small in magnitude and was not enhanced by repeated dosing. These results suggest that CYS is not an ideal tool for depletion of extrahypothalamic SRIF after sc or CNS administration and, moreover, raise serious questions about studies in which behavioral or endocrine alterations after CYS treatment were attributed to specific actions on SRIF-containing neurons. PMID- 2563245 TI - Effects of in vivo neuraminidase on the regulation of erythropoiesis. II: Modulation of erythroid colony formation by thymic regulatory cells. AB - Effects of the enzyme vibrio-cholerae neuraminidase (VCN) on the marrow-derived erythropoietic progenitor CFU-E and thymic regulatory cells were examined in vitro 1 and 24 h after i.v. injection of the enzyme. An in vivo enzymatic modification of bone marrow and thymic helper regulatory cell function occurs within 1 h after i.v. injection of VCN and results in suppression of both CFU-E colony formation and thymic helper cell function. These inhibitory effects of neuraminidase, however, are no longer detectable by 24 h after injection. More importantly, these inhibitory effects can be reversed by adding thymocytes from control animals to cocultures of enzymatically modified marrow or thymic regulatory cells. These findings: 1) suggest that regulatory cells from the bone marrow and thymus may be enzymatically modified in vivo in a reversible manner, suggesting a noncytotoxic effect of the enzyme on accessory cells, and 2) confirm the importance of sialic acid for the helper function but not for the suppressor function of thymocytes and CFU-E colony formation in vitro. PMID- 2563246 TI - The N-methyl-D-aspartate antagonists aminophosphonovaleric acid and MK-801 reduce anoxic damage to dentate granule and CA1 pyramidal cells in the rat hippocampal slice. AB - The effect of the N-methyl-D-aspartate antagonists, aminophosphonovaleric acid and MK-801, on irreversible transmission loss subsequent to anoxia was examined using the hippocampal slice preparation. A population spike was recorded from either the dentate granule cells or the CA1 pyramidal cells and the amplitude of this spike was compared before and 60 min following anoxia. After 10 min of anoxia the dentate granule cells recovered to 16 +/- 7% (mean +/- SE) of their preanoxic level when untreated and to 54 +/- 15% when treated with aminophosphonovaleric acid (APV). In slices treated with MK-801 the population spikes recorded from dentate granule cells recovered to 85 +/- 4% of their preanoxic level after 10 min of anoxia. Untreated CA1 pyramidal cells recovered to 8 +/- 3% of their preanoxic amplitude after 5 min of anoxia; they recovered to 59 +/- 6% when treated with MK-801 and 31 +/- 13% when treated with APV. The recovery of slices treated with the drugs was significantly different from that of untreated slices. ATP levels were measured in both the dentate and the CA1 region of slices. ATP in both regions fell less during anoxia when the slices were pretreated with either APV or MK-801. These differences between drug-treated and untreated tissue were significant with APV and MK-801. These differences between drug-treated and untreated tissue were significant with APV and MK-801 in dentate tissue after 10 min of anoxia and with MK-801 in CA1 tissue after 5 min of anoxia. This reduced fall in ATP during anoxia was accompanied by better physiological recovery after anoxia. We conclude that these NMDA antagonists provide protection against anoxic damage to dentate granule and CA1 pyramidal cells in this in vitro hippocampal preparation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563247 TI - Duplicated homeobox genes in Xenopus. AB - Multiple kinds of clones and restriction fragment polymorphisms are frequently encountered when analyzing genes of the tetraploid frog Xenopus laevis. Two types of cDNA clone have been isolated for homeobox gene 2. Analysis of their corresponding genomic clones confirmed the existence of clearly distinct restriction maps; in addition the nearby presence of two additional homeoboxes suggests that this region is homologous to the Hox 2 gene complex of mammals. We asked whether the genetic polymorphism in Xenopus results from increased allelic differences due to tetraploidy or from having duplicated Hox 2 complexes. Using X. laevis/Xenopus borealis interspecies hybrids we show that the two types of X. laevis homeobox gene 2 transcripts result from two different genetic loci. They cannot represent alleles of the same gene because they do not segregate independently in the F1 hybrid progeny. Most other X. laevis homeobox genes studied so far are also found in two versions. Thus X. laevis seems to have two homeobox genes, both of which are expressed, for each one present in mammals or other vertebrates. PMID- 2563249 TI - Ferritin H gene polymorphism in idiopathic hemochromatosis. AB - The authors studied the H ferritin restriction polymorphism in 83 hemochromatosis patients and 84 controls as well as in 19 nuclear families. No significant difference was found with the ten restriction enzymes used (HindIII, EcoRI, EcoRV, PvuII, BamHI, PstI, Bg/I, Bg/II, HincII, and TaqI). Hence, the genomic abnormality responsible for idiopathic hemochromatosis is not a major deletion of an H ferritin gene. A higher frequency of one HindIII fragment, although nonsignificant when the number of comparisons made is taken into account, was observed in the patients. This HindIII fragment hybridizes with the H ferritin probe and with a 28 S ribosomal probe, and its segregation with HLA haplotypes (hence its assignment to chromosome 6) is uncertain. Its possible meaning in the expression of the disease is discussed. PMID- 2563248 TI - Isolation of chromosome-21-specific DNA probes and their use in the analysis of nondisjunction in Down syndrome. AB - Thirteen single-copy, chromosome-21-specific DNA probes were isolated from a recombinant library made from flow-sorted chromosome 21 DNA and regionally mapped using a panel of somatic cell hybrids. Five probes mapped in the 21q21-q22.1 region, six to the 21q22.1-qter region, and one to each of the regions 21q22.1 q22.2 and 21q22.3. Two of these probes, one of which maps in the critical region for Down syndrome, have recently been shown to be expressed at high levels in Down syndrome brain tissue (Stefani et al. 1988). Following preliminary screening for restriction fragment length polymorphisms (RFLPs), five polymorphisms were discovered with four of the chromosome 21 DNA probes. A frequent MspI polymorphism detected by one of the probes was used in conjunction with four previously described polymorphic chromosome 21 probes to analyse the origin of nondisjunction in 33 families with a child or fetus with trisomy 21. The parental origin of the additional chromosome 21 was determined in 12 cases: in 9 (75%) of these it was derived from the mother and in the other 3 cases (25%) it was of paternal origin. Cytogenetic analysis of Q-banding heteromorphisms was informative in three of five families tested, and in each case the RFLP results were confirmed. The meiotic stage of nondisjunction was defined with confidence in five families, the results being obtained with pericentromeric RFLP or cytogenetic markers. Recombination between two nondisjoined chromosomes was demonstrated in one family and is consistent with the view that a lack of recombination between chromosome 21 homologues or failure of their conjunction is not the invariable cause of trisomy 21. PMID- 2563250 TI - Localization of the coagulation factor XIII B subunit gene (F13B) to chromosome bands 1q31-32.1 and restriction fragment length polymorphism at the locus. AB - In situ hybridization of tritiated cDNA probes for the gene for the B subunit of coagulation factor XIII localized the F13B locus to bands q31-q32.1 on human chromosome 1 and perhaps more precisely to sub-bands 1q31.2 or 1q31.3. Restriction fragment length polymorphisms (RFLPs) were detected with BglII, EcoRI and XbaI. Because the RFLPs detected with each of the three enzymes were concordant in every individual studied and since each showed a similar size difference, it was concluded that the RFLPs probably result from an insertion or deletion of length approximately 0.37-0.4 kb. PMID- 2563251 TI - The human cytochrome P450 CYP3 locus: assignment to chromosome 7q22-qter. AB - The cytochrome P-450s are a large multigene family of enzymes involved in the metabolism and detoxification of drugs and chemicals. Using a full length cDNA clone for the human nifedipine oxidase gene (CYP3) and a panel of human-rodent somatic cell hybrids, we have assigned this gene family to chromosome 7q22-qter. A search for RFLPs using this probe yielded no results. PMID- 2563252 TI - Confirmation of linkage disequilibrium between haplotype B (XV-2c, allele 1; KM 19, allele 2) and cystic fibrosis allele in the French population. AB - In 237 French families with cystic fibrosis (CF) restricted fragment length polymorphisms (RFLPs) were detected by two DNA probes, XV-2c and KM-19, which are tightly linked to the CF allele. As in other European populations linkage disequilibrium is found between the haplotype B (XV-2c, allele 1: KM-19, allele 2) and the CF allele. Linkage disequilibrium alters the probability that a person bearing a given haplotype is a carrier. PMID- 2563253 TI - Presymptomatic testing for Huntington's disease. A case complicated by recombination within the D4S10 locus. AB - Presymptomatic testing for Huntington's disease (HD) is possible through the use of restriction fragment length polymorphisms (RFLPs) at the closely linked D4S10 locus. Recombination between the HD and D4S10 loci will occur in 4%-5% of meioses, and is a well-recognised complication of predictive testing. Recombination between RFLPs within the D4S10 locus is a rare event and can usually be ignored. We report a case where such an intra-locus recombination frustrated attempts to predict the chance of a high-risk individual inheriting the HD gene. PMID- 2563254 TI - Molecular epidemiology of adhesin and hemolysin virulence factors among uropathogenic Escherichia coli. AB - The pap, prs, pil, and hly operons of the pyelonephritic Escherichia coli isolate J96 code for the expression of P, F, and type 1 adhesins and the production of hemolysin, respectively; the afaI operon of the pyelonephritic E. coli KS52 encodes an X adhesin. Using different segments of these operons as probes, colony hybridizations were performed on 97 E. coli urinary tract and 40 fecal clinical isolates to determine (i) the presence in the infecting bacteria of nucleotide sequences related to virulence operons, and (ii) the phenotypic properties associated with such sequences. Coexpression of P and F adhesins encoded by pap related sequences was detected more frequently among isolates from patients with pyelonephritis (32 of 49, 65%) than among those with cystitis (11 of 48, 23%; P less than 0.0001) or from fecal specimens (6 of 40, 15%; P less than 0.0001). Therefore, the expression of both adhesins appears to be critical in the colonization of the upper urinary tract. In contrast, afaI-related sequences were detected significantly more frequently among isolates from patients with cystitis, suggesting that this class of X adhesin may have a role in lower urinary tract infections. Urinary tract isolates differed from fecal isolates by a low incidence of type 1 adhesin expression among pil probe-positive isolates. hly-related sequences were only detected in pap probe-positive isolates. The frequency of hemolysin production among pap probe-positive isolates was not associated with a particular pattern of infection. The distribution of these virulence factors was similar in the presence or absence of reflux, indicating that structural abnormalities of the urinary tract did not facilitate colonization by adhesin-negative isolates. PMID- 2563255 TI - Structure and copy number of gene clusters related to the pap P-adhesin operon of uropathogenic Escherichia coli. AB - The structurally related pap and prs operons of the uropathogenic Escherichia coli isolate J96 encode a P and an F adhesin that mediate bacterial attachment to the human P blood group antigen and the Forssman antigen, respectively. Using probes prepared from different segments of the pap operon, Southern blot hybridizations were performed to characterize pap-related sequences of 30 E. coli clinical isolates expressing different adhesin phenotypes. Gene clusters encoding P and F adhesins displayed no restriction site polymorphism in sequences homologous to the papH, papC, and papD genes that encode proteins essential to the transport and polymerization of the subunits of the P-pilus adhesin. In contrast, pap-related genetic elements associated with a null phenotype either lacked homology to the papH, papC, and papD genes or displayed a restriction site polymorphism in this region. Sequences within and surrounding the J96 papG and prsG adhesin genes that determine the binding specificities to the P and F antigens, respectively, were not conserved. However, gene clusters encoding different binding specificities could not be distinguished based on such restriction site polymorphisms. The majority of clinical isolates had more than one copy of pap-related sequences that involved gene clusters similar to the J96 pap operon, as well as genetic elements that were related only to a part of this operon. The implications of this unexpected copy number polymorphism with respect to possible recombination events involving pap-related sequences are discussed. PMID- 2563256 TI - Involvement of the MN blood group antigen in shear-enhanced hemagglutination induced by the Escherichia coli F41 adhesin. AB - An adhesin from Escherichia coli F41 with an apparent subunit molecular weight of 28,000 daltons was isolated by using (NH4)2SO4 precipitation at pH 10 and Sephacryl S-500 gel filtration. The hemagglutination (HA) properties of the native high-molecular-weight adhesin were studied by using a viscometric assay, which provided a quantitative index of the degree of agglutination present as a function of time at a known rate of shear. Shear was found to enhance the degree of agglutination over a 20-min period. A strong, shear-enhanced HA was observed for all donors with the MM or MN blood type studied, but those with the NN blood type showed very little HA. In the microtiter HA assay, the selectivity of the adhesin for MM over NN erythrocytes was found to be dependent on pH and temperature. At 21 degrees C and pH 7.4, there was little difference in HA between the two blood types, but NN cells were progressively more weakly agglutinated than MM cells as the pH or the temperature was increased. Glycophorin A, which bears the M or N determinant, was isolated from individuals with the MM and NN blood types and was shown to be an effective inhibitor of the reaction, with the MM type being the more effective in both microtiter and viscometric assays. Acidic monosaccharides, particularly sialic acid, were also effective inhibitors of HA, although they were less potent on a molar basis than glycophorin. The adsorption isotherm of 125I-labeled adhesin was measured, and the binding was shown to be strongly inhibited by MM glycophorin and somewhat less strongly by NN glycophorin. Collectively, these data strongly suggest that glycophorin AM is a receptor for the F41 adhesin. PMID- 2563257 TI - Immunological studies of the disulfide bridge region of Pseudomonas aeruginosa PAK and PAO pilins, using anti-PAK pilus and antipeptide antibodies. AB - Pseudomonas aeruginosa is an opportunistic pathogen that attaches to host cells via their pili. The pilus of P. aeruginosa PAK consists of a polymer of a single subunit, pilin, which is a 144-residue polypeptide. The C-terminal end of this protein is semiconserved in a number of strains and contains a disulfide bridge. We have synthesized the C-terminal peptide PAK (128-144)-OH in both its reduced and oxidized forms and the analog PAK(A-129) (128-144)-OH, in which cysteine-129 was substituted by alanine. These three peptides were used to immunize rabbits and prepare antipeptide antisera. It was found that antipeptide antisera to reduced peptide (17-R) and to oxidized peptide (17-O) bound to native PAK pili and cross-reacted with strain PAO pili in direct enzyme-linked immunosorbent assay (ELISA) and immunoblot experiments. However, the antiserum to the peptide immunogen PAK(A-129)(128-144)-OH, which does not have the ability to form the disulfide bridge, did not bind to either PAK or PAO pili. Competitive ELISA experiments with reduced and oxidized peptides of Ac-PAK(128-144)-OH showed that there was no difference in binding between the two peptides for 17-R or 17-O immunoglobulin G. When immunoglobulin G from native PAK antipilus antiserum was used in competitive or direct ELISA experiments, there was also no preference in binding to reduced or oxidized Ac-PAK(128-144)-OH or to PAK(A-129)(128-144)-OH. This result showed that the disulfide bridge in Pseudomonas pili is not critical to the immunogenicity of this region. However, the disulfide bridge is important in the immunogenicity of the C-terminal peptide when preparing antipeptide antisera that are cross-reactive with pili from different strains, since only the disulfide bridge peptide antisera cross-reacted well with the PAO pili as shown by competitive ELISA, suggesting that this region could be an important candidate for development of a synthetic vaccine. PMID- 2563258 TI - Immunochemical identification and preliminary characterization of a nonfimbrial hemagglutinating adhesin of Bacteroides gingivalis. AB - A cell-bound hemagglutinating adhesin (HA-Ag2) of Bacteroides gingivalis was identified by crossed immunoaffinity electrophoresis as one of the common antigens of the species. A polyclonal antiserum with a restricted specificity for HA-Ag2 was produced by immunizing with the relevant immunoprecipitate excised from crossed-immunoelectrophoresis gels. The immunoglobulin G fraction of this monospecific antiserum inhibited hemagglutination. The antiserum was used against a cell surface extract of B. gingivalis in immunoblotting experiments, and we detected two antigens with apparent molecular masses of 33 and 38 kilodaltons in B. gingivalis ATCC 33277 and W83. Monoclonal antibody, C1.17, produced in another laboratory against B. gingivalis 381 and characterized as showing reactivity with a hemagglutinin of this strain (Y. Naito, K. Okuda, T. Kato, and I. Takazoe, Infect. Immun. 50:231-235, 1985), was also used to produce immunoblots of extracts of strains ATCC 33277 and W83. The apparent molecular masses of the major polypeptides recognized by monoclonal C1.17 in the immunoblots were the same as those detected by the polyclonal monospecific antiserum, i.e., 33 and 38 kilodaltons. Significantly, none of the polypeptides identified in this study corresponded to the polypeptide appearing in the 41- to 43-kilodalton region and identified by Yoshimura and co-workers (F. Yoshimura, K. Takahashi, N. Yoshinobu, and T. Suzuki, J. Bacteriol. 160:949-957, 1984) as the fimbrial protein characteristic of the species. Enzyme-linked immunosorbent assay inhibition experiments with the monospecific antiserum indicated that the cell surface extracts from strains ATCC 33277 and W83 were strong inhibitors, whereas the fimbria-enriched preparations from both strains failed to inhibit binding of antibodies to the cell surface antigens. As a whole, our study indicates that a nonfimbrial surface protein complex demonstrating erythrocyte-binding capacity, HA-Ag2, is common to three strains of B. gingivalis and is composed of at least two associated polypeptides with apparent molecular masses of 33 and 38 kilodaltons which share at least one antigenic determinant. PMID- 2563259 TI - Effect of alpha-1 and beta agonists on contraction of bovine retinal resistance arteries in vitro. AB - Contractile responses of bovine retinal arteries (BRA) (diameter: 179 +/- 9 micron, n = 25) to high K+, circumferential stretch and adrenergic stimulation were studied in vitro. BRA could be activated by rapid circumferential stretch. Under resting conditions, phenylephrine consistently activated BRA at the highest dose of the drug used (10(-5) M). During K+- and stretch-induced activation, significant contractile responses to phenylephrine appeared at lower doses (respectively, 3.10(-8) and 10(-6) M). Isoproterenol did not relax K+- and stretch-induced contractions. Therefore, (1) BRA probably can autoregulate through a myogenic mechanism on the basis of stretch; (2) during alpha 1 adrenergic stimulation, myogenic autoregulatory responses probably increase; (3) contractile responses to alpha 1 adrenergic stimulation are masked under resting conditions; and (4) BRA may not possess functional beta adrenergic receptors. PMID- 2563260 TI - Genetic and developmental regulation of the lipoprotein lipase gene: loci both distal and proximal to the lipoprotein lipase structural gene control enzyme expression. AB - We report here a study of the developmental and genetic control of tissue specific expression of lipoprotein lipase, the enzyme responsible for hydrolysis of triglycerides in chylomicrons and very low density lipoproteins. Lipoprotein lipase (LPL) mRNA is present in a wide variety of adult rat and mouse tissues examined, albeit at very different levels. A remarkable increase in the levels of LPL mRNA occurs in heart over a period of several weeks following birth, closely paralleling developmental changes in lipase activity and myocardial beta oxidation capacity. Large increases in LPL mRNA also occur during differentiation of 3T3L1 cells to adipocytes. As previously reported, at least two separate genetic loci control the tissue-specific expression of LPL activity in mice. One of the loci, controlling LPL activity in heart, is associated with an alteration in LPL mRNA size, while the other, controlling LPL activity in adipose tissue, appears to affect the translation or post-translational expression of LPL. To examine whether these genetic variations are due to mutations of the LPL structural locus, we mapped the LPL gene to a region of mouse chromosome 8 using restriction fragment-length polymorphisms and analysis of hamster-mouse somatic cell hybrids. This region is homologous to the region of human chromosome 8 which contains the human LPL gene as judged by the conservation of linked genetic markers. Genetic variations affecting LPL expression in heart cosegregated with the LPL gene, while variations affecting LPL expression in adipose tissue did not. Furthermore, Southern blotting analysis indicates that LPL is encoded by a single gene and, thus, the genetic differences are not a consequence of independent regulation of two separate genes in the two tissues. These results suggest the existence of cis-acting elements for LPL gene expression that operate in heart but not adipose tissue. Our results also indicate that two genetic mutations resulting in deficiencies of LPL in mice, the W mutation on chromosome 5 and the cld mutation on mouse chromosome 17, do not involve the LPL structural gene locus. Finally, we show that the gene for hepatic lipase, a member of a gene family with LPL, is unlinked to the gene for LPL. This indicates that combined deficiencies of LPL and hepatic lipase, observed in humans as well as in certain mutant strains of mice, do not result from focal disruptions of a cluster of lipase genes. PMID- 2563261 TI - End-to-end joining of taxol-stabilized GDP-containing microtubules. AB - By the use of the drug taxol, microtubules were assembled from tubulin that had GDP at its exchangeable nucleotide binding site. By means of dilution experiments and measurements of exchange of labeled subunits, it was determined that the rate of interchange of subunits between these microtubules and the solution is very slow: the upper limit of the dissociation rate constant was measured to be 0.2 subunit s-1 end-1. When they were broken into short pieces by gentle shearing, these microtubules were found to undergo a rapid subsequent spontaneous increase in length. This increase was attributed to end-to-end joining (also called annealing), because dynamic instability and other mechanisms involving either nucleotide hydrolysis or subunit interchange at the ends could be ruled out. To characterize the process, a diffusion-controlled joining mechanism was hypothesized, and a length-independent bimolecular rate constant, gamma, was defined. Length distributions were measured at a series of times after the initial shearing. By means of a novel iterative calculation, the best-fitting value of gamma was determined from the time-dependent changes in the length distributions. Fitting was carried out at each of three concentrations of microtubules. The resulting values show that end-to-end joining of microtubules is remarkably efficient and that gamma is concentration-dependent. PMID- 2563262 TI - Purification of a vanadate-sensitive ATPase from clathrin-coated vesicles of bovine brain. AB - Clathrin-coated vesicle acidification is mediated by an N-ethylmaleimide sensitive, vanadate-resistant proton-translocating ATPase. This enzyme is a 530 kDa hetero-oligomer which catalyzes ATP-dependent proton pumping when reconstituted (Xie, X. S., and Stone, D. K. (1986) J. Biol. Chem. 261, 2492 2495). We now report the purification of a second ATPase from bovine brain clathrin-coated vesicles which is inhibited by both N-ethylmaleimide (1 mM) and vanadate (10 microM). Localization of the ATPase to clathrin-coated vesicles was demonstrated by the precipitation of ouabain-resistant, vanadate-sensitive ATPase activity with anti-clathrin antibodies. The enzyme was solubilized with 0.1% polyoxyethylene 9-lauryl ether and has been purified 700-fold to a specific activity of 42 mumol of Pi.mg of protein-1.min-1. A molecular mass of 116 kDa was determined by centrifugation in sucrose gradients prepared in H2O and D2O, by high performance liquid chromatography using gel filtration, and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis performed under reducing conditions. The ATPase is unlike any known mammalian E1E2-type ATPase in that it is not inhibited by ouabain or [ethylenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA) and it is not activated by Na+, K+, or Ca2+. PMID- 2563263 TI - Prosomatostatin-derived antrin is present in gastric D cells and in portal blood. AB - The three widely distributed peptides derived from prosomatostatin are the original neurohormone somatostatin-14, somatostatin-28, and somatostatin-28(1 12), which are all derived from the COOH terminus of the precursor. Recently a decapeptide derived from the NH2 terminus of the prohormone has been identified in extracts of rat gastric antrum and named antrin. Data now show that in both rats and humans this new molecular form is concentrated in the D cell of the gastrointestinal mucosa together with somatostatin-28(1-12). The highest concentration of antrin immunofluorescent cells is located in the mucosa of the gastric antrum. Ultrastructural studies performed on pyloric D cells using the protein A-gold technique reveals that antrin is present in the same secretory granules as somatostatin-28(1-12) and detectable in one-third of all secretory granules. Acid extracts of rat hepatic portal plasma contain a peptide similar or identical to antrin, indicating that the antral decapeptide circulates in blood. PMID- 2563264 TI - Inhibition of gastrin gene expression by somatostatin. AB - Previous studies performed in this laboratory have demonstrated somatostatin containing cells in close proximity to gastrin cells in antral mucosa and have shown that somatostatin exerts a local regulatory effect on gastrin release. The present studies were directed to determine whether the effects of somatostatin on the antral gastrin cell involve pretranslational events. The effects of somatostatin on gastrin mRNA were determined by dot blot hybridization using a gastrin antisense RNA probe derived from human gastrin cDNA. Inclusion of somatostatin in the incubation medium caused a dose-dependent inhibition of steady-state gastrin mRNA. Conversely, when antral somatostatin was neutralized by the addition of specific somatostatin antibodies to the incubation medium, gastrin mRNA levels increased by 116 +/- 31% over control values (P less than 0.01). Northern blot hybridization of total antral RNA demonstrated a single major band with a molecular size of approximately 620 nucleotides, closely matching the predicted size of gastrin mRNA. The effect of somatostatin on the rate of gastrin gene transcription was examined using nuclear run-off transcription assays. Inclusion of antibodies to somatostatin in the incubation medium resulted in a 33.8 +/- 3.3% increase in gastrin gene transcriptional activity (P less than 0.01). These studies indicate that, in addition to its established effect on peptide release, somatostatin exerts inhibitory effects on antral gastrin cells at the pretranslational level. Although this inhibition appears to occur in part at the gene transcriptional level, the results also indicate that somatostatin may affect posttranscriptional processing of gastrin mRNA. PMID- 2563266 TI - Replication of the human immunodeficiency virus 1 and impaired differentiation of T cells after in vitro infection of bone marrow immature T cells. AB - HIV-1 infection in vitro of normal bone marrow mononuclear cells (BMMC) depleted of mature T cells was studied. BMMC depleted of either CD3, CD2, or both could replicate HIV-1 irrespective of the presence of macrophages/monocytes. Infected bone marrow cells were shown to differentiate during the culture into CD3+, CD4+, CD8+, and CD1+ cells, whereas noninfected BMMC gave rise to CD3+, CD4+, and CD8+ cells. Moreover, 9-14% of the cells also expressed the viral proteins p24 and gp120 on their surface. Double staining studies revealed that 72 and 83% of the CD4+ cells expressed the gp120 and p24, respectively, suggesting that virus replication occurred in CD4+ cells. T cell colony growth from infected BMMC, either unfractionated or depleted of mature T cells, was impaired in a time dependent manner, and the differentiation capacity of T cell precursors was abnormal. Colony cells displayed an immature cell phenotype (CD1+ cells) and the viral proteins gp120 and/or p24 could also be detected on CD1+ cells. In addition, pooled colony cells derived from infected CD2- and CD3-depleted BMMC could infect normal mitogen-activated lymphocytes in coculture experiments. These findings strongly suggest that HIV-1 can infect immature bone marrow T cells and be transmitted to the progeny, but the massive viral replication occurs only when the cells differentiate toward CD4+ cells. PMID- 2563267 TI - Correspondence between high affinity 125I-neurotensin binding sites and dopaminergic neurons in the rat substantia nigra and ventral tegmental area: a combined radioautographic and immunohistochemical light microscopic study. AB - Several lines of anatomical, biochemical, and pharmacological evidence have suggested that specific high affinity neurotensin binding sites are associated with dopamine-containing neurons in the rat ventral tegmentum. In the present study we confirmed and quantified the extent of this association by combining monoiodinated neurotensin radioautography and tyrosine hydroxylase immunohistochemistry on adjacent 5-10 microns-thick midbrain sections. We found that 95-100% tyrosine hydroxylase-immunoreactive neurons detected in all subdivisions of the substantia nigra (pars compacta, pars lateralis, and pars reticulata) exhibited intense 125I-neurotensin labeling in adjacent light microscopic radioautographs. Tyrosine hydroxylase-positive dendrites radiating downward from compacta neurons were also heavily labeled throughout the pars reticulata. In the paranigral subdivision of the ventral tegmental area, silver grains were evenly distributed over neuropil and perikarya and therefore could not be readily attributed to any given tyrosine hydroxylase-positive element. In contrast, within the parabrachial pigmentous subdivision of the ventral tegmental area, 80-90% of the tyrosine hydroxylase-immunoreactive somata and proximal processes were clearly in register with 125I-neurotensin labeled cells. Finally, all tightly packed TH-positive neurons in the interfascicular nucleus showed intense 125I-neurotensin labeling. The vast majority of the neurotensin binding sites observed in the ventral midbrain tegmentum were of the high affinity, physiologically active receptor type since levocabastine, a selective blocker of the low affinity neurotensin binding component, had minimal effect on the binding density in any of the midbrain regions examined. The present results demonstrate an extensive overlap between specific, high affinity neurotensin binding sites and dopamine perikarya and dendrites in the rat ventral tegmentum, and thereby provide a direct anatomical substrate for observed neurotensin-dopamine interactions in the mesocorticolimbic and nigrostriatal projection systems. PMID- 2563268 TI - Catecholamine innervation of the human cerebral cortex as revealed by comparative immunohistochemistry of tyrosine hydroxylase and dopamine-beta-hydroxylase. AB - The organization of the cortical monoamine systems, dopamine (DA), and noradrenaline (NA), which have been studied extensively in the rat and more recently in the monkey, had not yet been investigated directly in the human brain. We report here the first systematic account of the regional and laminar distributions of the catecholamine fibers in the human cerebral cortex, using immunohistochemistry of the catecholamine biosynthetic enzymes, tyrosine hydroxylase (TH), and dopamine-beta-hydroxylase (DBH) in 13 cytoarchitectonic areas (4, 6, 9, 3b, 5, 40, 17, 18, 23, 24, 29, insula, and hippocampus) sampled postmortem. The noradrenergic (NA) innervation, mapped with DBH-immunoreactivity (DBH-IR), displayed a characteristic density gradient in the neocortex (highest in the primary sensorimotor areas, decreasing rostrally and caudally) that contrasted with the more uniform density in the limbic cortices (24, 23, 29, insula, hippocampus). NA axons were present in all cortical layers and were least numerous in layer I. The DBH-IR fibers were only partly TH-immunostained (10-50%, on double-labeled sections), suggesting a heterogeneity of the cortical NA axons. The putative dopaminergic (DA) fibers were identified by comparing alternate or double-immunolabeled (DBH-TH) sections, as the TH-IR fibers which contain no DBH IR. A DA-like innervation was present in all cortical areas, with major regional differences in density and laminar distribution, which closely paralleled cytoarchitectural buildups: 1) the DA-like innervation was densest in the agranular areas, primary and secondary motor areas, anterior cingulate, and insula; it distributed throughout layers I-VI; 2) density was lower in the granular cortices, areas 9 (prefrontal cortex), 23, 3b, 5, 40, and 18, displaying a bilaminar pattern in layers I and V-VI. In all areas, DA-like fibers were most abundant in the molecular layer, with a predominant distribution in its deepest part. Convoluted and coily fibers represented a unique morphologic aspect of the CA innervation in the human cortex. These findings are in agreement with findings in nonhuman primates and demonstrate major evolutionary changes in the organization of the cortical aminergic input as compared with rodents. The most striking features are the expansion of the DA innervation to the whole cortex and the peak of highest density in the motor areas. The regional differentiation of NA innervation is also accentuated. Slight differences were found in the laminar distributions of the amines in humans and primates. These data seem quite promising and open new research fields in neurologic and psychiatric diseases. PMID- 2563265 TI - Effects of acute ischemia in the dog on myocardial blood flow, beta receptors, and adenylate cyclase activity with and without chronic beta blockade. AB - We ligated the left anterior descending coronary artery for 1 or 2 h in 31 purebred beagles. We did not detect any changes in beta-adrenergic receptor density or affinity when normal and ischemic zones were compared, either in the subendocardium or in the subepicardium. In the ischemic zones, there was a significant decline in all measures of adenylate cyclase activity, including activity mediated by the beta-adrenergic receptor. By contrast, after chronic beta-adrenergic blockade (1.5 mg/kg propranolol i.v. twice daily for 7 d), there was an increase in adenylate cyclase activity stimulated by (-)-isoproterenol relative to adenylate cyclase activity stimulated by guanyl-5'imidodiphosphate (GppNHp) in both normal and ischemic tissue, suggesting that one effect of chronic beta blockade may be to enhance coupling between the stimulatory guanine nucleotide regulatory protein (Gs) and the beta-adrenergic receptor, despite a reduction in the number or function of Gs units. Chronic beta blockade also led to up regulation of beta-adrenergic receptor density in subepicardial regions. After 20 min of reperfusion following 2 h of ischemia, adenylate cyclase activity tended to return to control levels, particularly in the subepicardium, where (-) isoproterenol-stimulated adenylate cyclase activity was not different from normal myocardium. We conclude that chronic beta-adrenergic blockade may have beneficial effects during prolonged episodes of myocardial ischemia by preserving signal transduction mediated by the beta-adrenergic receptor. PMID- 2563269 TI - Association of class II DNA restriction fragments with responsiveness to Ambrosia artemisiifolia (short ragweed)-pollen allergen Amb a V in ragweed-allergic patients. AB - Human IgE and IgG antibody responsiveness to the short ragweed-pollen allergen Amb a V (formerly known as Ra5) has been found to be strongly associated with HLA D specificities Dw2 and DR2 in ragweed-allergic white individuals. To study the molecular basis of these associations, restriction fragment length polymorphism (RFLP) mapping was performed on a group of 45 white ragweed-allergic patients with full-length HLA-DR beta, DQ beta, and DQ alpha cDNA probes. The data on 41 of these subjects were used for the purposes of statistical analysis. With the DR beta probe, we found that the presence of three polymorphic restriction fragments correlated with responsiveness to Amb a V and with the DR2 specificity, namely, a 6.5 kb Eco RI fragment, a 9.4 kb Hind III fragment, and a 2.2 kb Hind III fragment. The presence of four fragments detected with the DQ beta probe correlated with responsiveness to Amb a V and with Dw2 specificity: a 2.3 kb Eco RI fragment, a 13.0 kb Pst I fragment, a 2.9 kb Taq I fragment, and a 5.2 kb Eco RV fragment. The DR beta Eco RI 6.5 kb and the DQ beta Eco RI 2.3 kb fragments were studied in detail; the concordant presence of these fragments was even more strongly associated with responsiveness to Amb a V. Fifteen of 17 responders had both fragments, whereas only one of 24 nonresponders had both fragments (p = 5 x 10(-7). This is the first time that such an association has been found between a person's immune response to a well-defined antigen and a set of HLA class II DNA restriction fragments. PMID- 2563270 TI - Rimiterol and the prevention of exercise-induced asthma. AB - The potential for rimiterol to protect athletes from exercise-induced asthma (EIA) has not been fully established. Ten athletes with asthma (15 to 30 years of age) undertook 8 minutes of submaximal exercise (80% of anaerobic threshold) on the treadmill ergometer, once after inhaling rimiterol and once after inhaling a placebo. Treatment with all bronchodilator drugs was stopped for the 12 hours preceding each exercise test. Two puffs (400 micrograms) of rimiterol or placebo were administered in a double-blind crossover manner 2 minutes before each exercise test. Lung function measurements were made before exercise and immediately, 5, 10, 15, 20, 25, and 30 minutes after completion of exercise. The results of a two-way analysis of variance revealed significant (p less than 0.01) difference in the FEV1 scores obtained after rimiterol inhalation and placebo inhalation, 5, 10, 15, 20, 25, and 30 minutes after cessation of exercise. After inhalation of rimiterol, there were no significant changes in FEV1. After inhaling the placebo, significant reductions (p less than 0.01) in FEV1 occurred after cessation of exercise (5, 10, 15, and 20 minutes). All subjects exhibited EIA after placebo, and none after rimiterol. The mean maximum drop after exercise in FEV1 after inhalation of rimiterol (2.807 +/- 5.55) and placebo (24.54 +/- 8.4) was significantly different (t = 6.849). It was concluded that inhalation of rimiterol 2 minutes before exercise afforded significant protection from EIA in all subjects tested. PMID- 2563271 TI - T cell receptor beta-chain genes in the rat. Availability and pattern of utilization of V gene segments differs from that in the mouse. AB - We have constructed a specifically primed thymus cDNA library to study the V region repertoire of the TCR beta-chain in the laboratory rat. From this library, we have isolated and sequenced 14 clones that hybridize with beta-chain constant region probes, eight of which contain V gene segments. Two of these eight contain a V gene segment whose sequence is 85% identical to mouse V beta 14, which has been estimated to be present in mouse thymocyte cDNA at a frequency of only 1.2%. Of the remaining six, four appear to be homologues of known mouse V beta sequences, and average 88% identity with their mouse counterparts. Two additional rat V regions have no homologue yet identified in the mouse, showing only 64 and 73% identity with the most closely related mouse sequences; at least one of these is present and polymorphic in the mouse genome, and may be located near one end of the V beta deletion known in the SWR strain. The availability and pattern of V beta gene segment expression in rat thymus appears, therefore, to differ significantly from the mouse. Two cDNA clones represent germline transcripts which show a J/C splice (one of which is defective) but no D/J rearrangement; two different J segments are used, indicating the presence of promoter activity 5' to these two J segments. In addition, by using V beta-specific probes derived from our cDNA clones, we have analyzed the distribution of V beta alleles among 10 inbred strains of rat. We find that rat V beta gene segments occur predominantly in single member subfamilies, and present evidence for the existence of three TCR V beta haplotypes defined by RFLP. PMID- 2563272 TI - Mapping of immunogenic regions of human T cell leukemia virus type I (HTLV-I) gp46 and gp21 envelope glycoproteins with env-encoded synthetic peptides and a monoclonal antibody to gp46. AB - Antigenic sites on human T cell leukemia virus type I (HTLV-I) gp46 and gp21 envelope glycoproteins that are immunogenic in man were studied with envelope gene (env)-encoded synthetic peptides and a mAb to HTLV-I gp46 envelope glycoprotein. Antibodies in 78% of sera from HTLV-I seropositive subjects reacted with synthetic peptide 4A (amino acids 190 to 209) from a central region of HTLV I gp46. Human anti-HTLV-I antibodies also bound to synthetic peptides 6 (29% of sera) and 7 (18% of sera) from a C-terminal region of gp46 (amino acids 296 to 312) and an N-terminal region of gp21 (amino acids 374 to 392), respectively. mAb 1C11 raised to affinity-purified HTLV-I gp46 reacted with gp46 external envelope glycoprotein and gp63 envelope precursor in immunoblot assay and also bound to the surface of HTLV-I+ cells lines HUT-102 and MT-2. Antibody 1C11 did not react with HTLV-II or HIV-infected cells or with a broad panel of normal human tissues or cell lines. In competitive RIA, anti-gp46 antibody 1C11 was inhibited from binding to gp46 either by antibodies from HTLV-I seropositive subjects or by HTLV I env-encoded synthetic peptide 4A, indicating that 1C11 bound to or near a site on gp46 within amino acids 190 to 209 also recognized by antibodies from HTLV-I seropositive individuals. When tested in syncytium inhibition assay, mAb 1C11 did not neutralize the infectivity of HTLV-I. Thus, HTLV-I infection in man is associated with a major antibody response to a region of gp46 within amino acids 190 to 209 that is on the surface of virus-infected cells. PMID- 2563273 TI - Mitogenic stimulation of human B lymphocytes by the mannose-specific adhesin on Escherichia coli type 1 fimbriae. AB - Escherichia coli type 1 fimbriae contain in association with the major structural protein a lectin-like adhesin moiety that mediates attachment of E. coli to mannose-containing receptors on the surface of host cells. We have investigated the lymphocyte mitogenic activity of this mannose-specific adhesin by comparing the ability of purified wild type type 1 fimbriae containing the adhesin and mutant type 1 fimbriae lacking the adhesin to stimulate proliferation in human lymphocytes. Both fimbriae stimulated a peak of proliferation at 8 days whereas only the wild type fimbriae stimulated an additional peak of proliferation occurring at 3 days. Proliferation at 3 days but not at 8 days could be blocked by the addition of alpha-methyl-D-mannoside. Neonatal lymphocytes from umbilical cord blood responded to both wild type and mutant fimbriae in a fashion similar to adult cells. Stimulation of separated T and non-T cell populations indicated that the proliferation seen at 3 days was solely due to non-T cells whereas the 8 day response was due to T cell proliferation. The addition of gamma-irradiated T cells did not appear to enhance the 3-day response of the non-T cells. However, the 8-day response by T cells was dependent on the presence of gamma-irradiated non-T cells. In cultures of unseparated cells, wild type fimbriae stimulated more than 75% of the B cells to enter the S and G2 phase at 3 days whereas at 8 days cycling T cells were present in both wild type and mutant fimbriae-stimulated cultures. Taken together, our observations suggest that the adhesin molecule stimulates a polyclonal mitogenic response in B cells that peaks at 3 days, and other structural components of the fimbriae are responsible for evoking an 8-day (probably immune) response in T cells. PMID- 2563274 TI - Drug therapy for hypertension in the elderly. AB - Essential hypertension is a major health care problem in the elderly and requires effective treatment to reduce morbidity and mortality. The traditional stepped care approach to therapy consisted of diuretics; sympatholytic agents, or beta blockers for all age groups. Indeed, initial therapy with these agents is effective in 50 to 60 percent of elderly patients but may produce adverse effects. A high incidence of adverse responses, including sexual dysfunction and central nervous system impairment, has been reported with diuretic or beta blocker therapy, and a reduction in several measures of quality of life has been noted during therapy with methyldopa or propranolol. Administration of an angiotensin-converting enzyme (ACE) inhibitor is as effective as the traditional stepped-care approach without producing the ill effects associated with diuretics, sympatholytics, or beta-blockers. The combination of an ACE inhibitor with a diuretic produces additive antihypertensive effects while minimizing diuretic-induced metabolic alterations. Orthostatic hypotension with the first dose can be minimized by making sure that patients are not hypovolemic from previous diuretic therapy. Nevertheless, in controlled trials, the combination of ACE inhibitor and diuretic has been effective in up to 85 percent of patients. In addition, the use of ACE inhibitors may be beneficial in the hypertensive patient with concomitant congestive heart failure. Most important, the patient's quality of life is maintained during therapy with an ACE inhibitor alone or in combination with a diuretic. Thus, an ACE inhibitor plus a diuretic is a valuable alternative to traditional antihypertensive therapy in elderly patients. PMID- 2563275 TI - Flow cytometric analysis and isolation of permeabilized dopamine nerve terminals from rat striatum. AB - Fluorescence-activated cell sorting (FACS) was used to identify and isolate permeabilized dopaminergic nerve terminals from rat striatum based on immunofluorescent labeling of tyrosine hydroxylase (TH). Striatal synaptosomes were permeabilized by fixation with modified Zamboni fluid. A highly fluorescent subpopulation of particles was detected by FACS following sequential incubation with a monoclonal antibody to TH (LNC 1) and a fluorescein-conjugated secondary antibody. After correcting for nonsynaptosomal particles present in the synaptosomal fraction, analysis of these data suggested that approximately 12-15% of striatal synaptosomes were dopaminergic, consistent with previous estimates. Specific labelling by LNC 1 was decreased if synaptosomes were prepared from rats that had received intraventricular injections of 6-hydroxydopamine. The decrease in labeling was highly correlated with the extent of the lesion as determined by measurement of striatal dopamine levels, suggesting that LNC 1-labeled synaptosomes were derived from nigrostriatal dopamine terminals. In order to verify that LNC 1-labeled synaptosomes were enriched for TH, equal numbers of labeled and control synaptosomes were isolated by FACS and analyzed by SDS-PAGE. LNC 1-labeled synaptosomes were shown by Western blot techniques to be enriched 6 fold for TH compared with control synaptosomes, suggesting that the labeled population consisted almost entirely of dopaminergic synaptosomes. PMID- 2563276 TI - Secretin and vasoactive intestinal peptide activate tyrosine hydroxylase in sympathetic nerve endings. AB - Secretin and vasoactive intestinal peptide (VIP) are known to stimulate tyrosine hydroxylase (TH) activity acutely in the rat superior cervical ganglion (SCG). Because TH-containing neurons in the SCG innervate the iris, submaxillary gland, and pineal gland, we examined the effects of secretin and VIP in these 3 autonomic end organs in vitro. Both peptides stimulated TH activity in each tissue. These stimulations resembled those in the SCG in that (1) secretin displayed a higher potency than VIP in all 3 end organs, (2) the peptide effects were unchanged when calcium was excluded from the incubation medium, and (3) they were mimicked by activators of the cyclic adenosine monophosphate (cAMP) pathway. These findings indicate that secretin and VIP can regulate transmitter metabolism in both the cell bodies and axon terminals of neurons originating in the SCG. Furthermore, the data raise the possibility that catecholamine synthesis in sympathetic nerve terminals is modulated by peptides released by other, nearby nerve endings. PMID- 2563277 TI - Bursting induces persistent all-or-none EPSPs by an NMDA-dependent process in piriform cortex. AB - Burst responses to stimulation of excitatory fiber tracts in olfactory cortex slices after removal of extracellular Mg2+ or decreases in extracellular Cl-, resulted in long-lasting changes in response properties of neurons following a return to normal bathing medium. After bursting activity, the response of pyramidal cells to stimulation of afferent or associational fiber systems consisted of the normal graded depolarizing postsynaptic potential and a new, high-amplitude depolarizing potential that followed the graded potential at a variable latency. The new late potential had a waveform that resembled the initial graded response, but it occurred in an all-or-none fashion with a discrete threshold and persisted for many hours. Threshold for the late potential was similar for different cells in the same slice and was not affected by intracellular current injection, indicating that a synchronized interaction among a large number of cells is involved in its generation. Properties of the late potential indicate that it is an EPSP. NMDA receptor antagonists (APV and ketamine) had little effect on the late potential but prevented its development if present during bursting activity. The possible relevance of these findings to the study of the neuronal substrate for long-term memory and epilepsy is discussed. PMID- 2563278 TI - Neuroleptic-sensitive binding sites in the nigrostriatal system: evidence for differential distribution of sigma sites in the substantia nigra, pars compacta of the cat. AB - The sigma site is a recently described binding site to which dextrorotary isomers of certain psychotomimetic benzomorphans bind specifically. Classical and nonclassical neuroleptics also have high affinity for the sigma site, including neuroleptics known to bind to the D2 dopamine receptor. The affinity of some D2 binding neuroleptics for the sigma site has raised the possibility that certain important effects of antipsychotic drugs may relate to the sigma site. Left unresolved has been the question of how these actions could relate to dopaminergic systems. To explore this issue we carried out an autoradiographic binding study of the distribution of the sigma-selective ligand 3H-DTG in the nigrostriatal system. We report here that haloperidol-displaceable 3H-DTG binding sites are densely concentrated in an anatomically discrete subdivision of the cat's substantia nigra pars compacta. This zone, identifiable as the striosome projecting densocellular zone of the pars compacta, also shows heightened D2 related ligand binding but has reduced D1-related ligand binding relative to other parts of the nigral complex. This evidence suggests that sigma-mediated interactions with dopaminergic systems may occur in the substantia nigra pars compacta and that the functional effects of these interactions may influence the nigrostriatal projection to striosomes differentially. PMID- 2563279 TI - Dynamics of alpha-tubulin deacetylation in intact neurons. AB - The majority of the alpha-tubulin in cultured neurons is acetylated (Black and Keyser, 1987). The present studies examine the relationships of the acetylation and deacetylation reactions to tubulin assembly and disassembly in intact neurons. Extraction assays which separate assembled and unassembled tubulin pools reveal that greater than or equal to 99% of the total acetylated, as well as newly acetylated, tubulin is cytoskeletal associated. Treatment of neurons with depolymerizing drugs results in a progressive decrease in the levels of total tubulin in polymer and a corresponding increase in the levels of soluble tubulin. These drugs also cause a progressive decrease in the levels of acetylated alpha tubulin in polymer that closely parallels in rate and extent that of total alpha tubulin. However, there is no corresponding increase in soluble acetylated tubulin. Because the total levels of alpha-tubulin remain unchanged during drug treatment, the decrease in levels of acetylated alpha-tubulin during depolymerization must reflect its rapid conversion to nonacetylated alpha tubulin. These findings suggest alpha-tubulin is acetylated in the polymeric form and that deacetylation is closely coupled to depolymerization. The close coupling between alpha-tubulin deacetylation and depolymerization provided a means of estimating the rate at which subunits cycle off microtubules in intact neurons. Acetate turnover on tubulin in intact neurons was determined both by pulse-chase protocols with 3H-acetate and by measuring the loss of acetylated subunits (using quantitative immunoblotting) under conditions of net microtubule depolymerization induced by colchicine. Both methods yielded similar results. Acetate turnover occurred biphasically; 30-50% of the acetate on tubulin turns over with a t1/2 of 1.5-2 hr, and the remaining half or more turns over with a t1/2 of 5-10 hr. We suggest that these kinetically distinguishable pools of acetylated alpha-tubulin reflect distinct pools of acetylated microtubules that differ in their average rates of subunit turnover. PMID- 2563280 TI - Aplysia synaptosomes. II. Release of transmitters. AB - A subcellular fraction (P3) from Aplysia is enriched in synaptosomes (Chin et al., 1988) and is capable of accumulating 5-HT and choline. At an external 3H-5 HT concentration of 1.8 microM, the P3 fraction took up 0.12 nmol/mg protein in 30 min. Uptake was dependent on external Na+. Electron microscopic autoradiography showed that much of the accumulated 3H-5-HT is localized to synaptosomes. At 0.5 microM 3H-choline, P3 took up 0.11 nmol/mg protein in 30 min and converted 40% to 3H-ACh. This synaptosomal fraction was also capable of releasing transmitter. After 3H-5-HT or 3H-choline was taken up, P3 released about 5% of the total radioactive transmitter in a Ca2+-dependent manner during a 30 sec exposure to a depolarizing concentration of K+ (100 mM). Identified, prelabeled synaptosomes were prepared by injecting 3H-choline into the large cholinergic neuron L10. The abdominal ganglia containing the injected cells were then fractionated, yielding synaptosomes containing radioactivity derived from L10. After this synaptosomal fraction was exposed to high K+, 2% of the radioactivity was released in a Ca2+-dependent manner. This release was completely blocked by 0.1 mM histamine, a modulatory transmitter that has previously been shown to cause presynaptic inhibition in L10. PMID- 2563281 TI - Separation of quisqualate- and kainate-selective glutamate receptors in cultured neurons from the rat superior colliculus. AB - The aim of the present study was to identify and characterize the receptors and ionic channels mediating the compound response of tectal neurons to exogenous L glutamate (Glu). Particular attention was paid to the question of whether separate receptors and channels exist for quisqualate (QA) and kainate (KA) and, if so, whether binding to one of these receptors would modify the response elicited through the other. Neurons were dissociated from the superficial gray layer of the superior colliculus from E21 or P1 rats. Between days 14 and 21 in vitro, responsiveness of tectal neurons to Glu and related substances was tested by recording the whole-cell currents induced by rapid superfusion with drug containing salt solutions. Our experiments showed that tectal neurons express at least 3 distinct types of receptors for acidic amino acids. KA-activated currents (I(KA)) differ from QA-activated currents (I(QA)) in their dose-response characteristics, desensitization patterns, selective blockade with kynurenic acid and suppression by elevated [Ca2+]o, I(KA), but not I(QA), is significantly reduced by low levels of [Cl-]o, and the [Cl-]o-dependent shift of the reversal potential for I(KA) suggests that KA promotes a conductance decrease for Cl-. Such an effect has been ascribed to APB-receptors, but L-2-amino-4 phosphonobutyrate (APB) itself failed to induce current responses in tectal neurons. KA was without effect when administered together, and in equimolar concentrations, with QA. The block of I(KA) was, however, surmounted by applying KA at considerably higher concentrations. It is concluded that QA acts as a low affinity competitive antagonist at the KA site and as a high-affinity agonist at its own receptor. The response to the endogenous ligand Glu reflects properties of all receptors. QA and KA receptors account for 20-30% (QA) and 49-82% (KA) of the compound current elicited with 100 microM Glu. These results indicate that binding of Glu does not, in contrast to QA, produce any significant suppression of the KA-receptor-mediated current component. PMID- 2563282 TI - Effects of locus coeruleus lesions on auditory, long-latency, event-related potentials in monkey. AB - It has previously been demonstrated that monkeys exhibit certain event-related potential (ERP) components showing latency, polarity, and contingency similarities to those observed in humans. In the present study, monkey P300-like components were studied in order to evaluate the hypothesis that the noradrenergic locus coeruleus (LC) system participates in their generation or modulation. ERPs were recorded from untrained squirrel monkeys (Saimiri sciureus) twice a week for 4 weeks before and after bilateral LC lesions and interruption of dorsal bundle (DB) fibers. Stimuli consisted of 2 and 6 kHz tone pips (40 msec duration, 60 dB above nHL) presented once a second in random order. In most sessions, one tone constituted 90% of the stimuli and the other tone 10%, while in some sessions tones were made equiprobable to test the effects of manipulating stimulus probability. LC and DB lesions were made by first localizing the nucleus and creating an electrolytic lesion. Then, the electrode was placed at the anterior pole of the nucleus and a knife cut effected. The extent of damage to LC perikarya and ascending axons was assessed by reconstructing lesions from Nissl stained sagittal sections through the brain stems. The effect of lesions on cortical noradrenergic axons was immunohistochemically verified utilizing antisera directed against dopamine-B-hydroxylase and tyrosine hydroxylase to label noradrenergic and dopaminergic axons, respectively. The prelesion ERP results replicated previous findings of P300-like components recorded in response to low-probability tones. The postlesion ERP data indicated that following damage to LC cell bodies, combined with interruption of histochemically detectable ascending noradrenergic axons, monkey P300-like potentials exhibited decreased areas, altered brain-surface distribution, and reduced sensitivity to stimulus probability. The correlation between the extent of cell body lesions and percentage reduction in the magnitude of P300-like responses was significant. However, interruption of DB fibers alone did not have similar effects. Neither type of lesion had any effect on amplitudes, latencies, or brain-surface distributions of P52, P172, or N250-900. There was, however, a significant effect on N106. Stimulus probability effects on the frontally distributed P52 and N106 were not altered by the lesions. These data support the hypothesis that the integrity of the LC nucleus and its ascending fibers is important in the generation and modulation of surface-recorded P300-like activity. PMID- 2563283 TI - Flow cytometric analysis of rat striatal nerve terminals. AB - Methods were developed for the analysis and isolation of striatal nerve terminals (synaptosomes) using fluorescence-activated cell sorting (FACS). Comparison of the light-scattering properties of synaptosomal and mitochondrial fractions indicated that particles in the synaptosomal fraction were generally larger and more sensitive to hypotonic lysis, consistent with results obtained by other methods of analysis. FACS analysis using indirect immunofluorescence techniques indicated that approximately 84% of the synaptosomal fraction was labeled by monoclonal antibody (mAb) A2B5 and thus appeared to be of neuronal origin. After permeabilization, between 5 and 10% of the particles were labeled by a mAb to glial fibrillary acidic protein, suggesting that they were derived from astrocytes. A fluorescent voltage-sensitive dye (VSD) was used to distinguish intact synaptosomes from free mitochondria (only the former maintain a membrane potential under the present experimental conditions). Approximately 83% of the synaptosomal fraction exhibited increased fluorescence after incubation with the VSD; furthermore, the fluorescence signal decreased in response to depolarizing agents (elevated potassium and veratridine). A portion of the mitochondrial fraction responded similarly, consistent with the presence of contaminating synaptosomes. Analysis of synaptosomal labeling by 11 fluorescein-conjugated plant lectins indicated that striatal nerve terminals differ significantly in their cell surface glycoconjugates. Subpopulations of synaptosomes defined on the basis of lectin binding were collected by FACS onto filters and probed with a mAb to tyrosine hydroxylase (TH) using Western blot techniques. While subpopulations exhibited different amounts of TH immunoreactivity, none of the lectins appeared to recognize TH-positive (i.e., dopaminergic) synaptosomes exclusively. These findings demonstrate that synaptosomes can be characterized and isolated for further study based on FACS analysis of properties such as size, membrane potential, and the presence of intracellular or cell surface molecules. PMID- 2563284 TI - Brain cell and tissue recovery in rats made deficient in n-3 fatty acids by alteration of dietary fat. AB - Rats were fed a purified diet containing either 1.5% sunflower oil [940 mg linoleic acid [18:2(n-6)]/100 g diet; 6 mg alpha-linolenic acid [18:3(n-3)]/300 g diet] or 1.9% soybean oil [940 mg 18:2(n-6)/100 g diet; 130 mg 18:3(n-3)/100 g diet]. In all cases and tissues examined 22:6(n-3) was lower and 22:5(n-6) was higher in rats fed sunflower oil than in rats fed soybean oil. Levels of 22:4(n 6) and 20:4(n-6) were largely unaffected. Expressed as a percentage of that in soybean oil-fed rats, 22:6(n-3) in sunflower oil-fed rats was as follows: neurons, 49; astrocytes, 47; oligodendrocytes, 10; lung, 27; testes, 32; retina, 36; liver, 35 and kidneys, 45. Ten wk after the change in diet of 60-d-old rats from one containing sunflower oil to one containing soybean oil, the fatty acid composition of the brain cells had not reached control values, e.g., that obtained in animals continuously fed soybean oil; 22:6(n-3) was 77, 65 and 80% of control levels for astrocytes, oligodendrocytes and neurons, respectively. In contrast, the recovery measured by the decay of 22:5(n-6) was complete within 10 wk. For 22:6(n-3), it took approximately 2 wk for liver and kidney to recover to the control value, 3 wk for lung, 6 wk for retina and 10 wk for testes. The decrease of 22:5(n-6) was rapid: the control values were reached within 2 wk for kidney, liver and lung and within 6 wk for retina.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563285 TI - Dietary biotin requirements of young rainbow trout (Salmo gairdneri) determined by weight gain, hepatic biotin concentration and maximal biotin-dependent enzyme activities in liver and white muscle. AB - The objective of the present work was to determine the dietary biotin requirements of young, rapidly growing rainbow trout according to independently measured parameters. Two experiments were conducted with a purified diet which had a basal biotin level of 0.01-0.02 mg/kg. A third study was done with a nonpurified diet with or without a supplement of 1.0 mg biotin/kg. Each study was initiated with fry weighing less than 2 g/fish, and was continued for 16-20 wk at 15 degrees C. The first experiment, a cross-over design with pair-feeding, showed that the unsupplemented purified diet produced a biotin-specific deficiency condition in the trout. Dietary requirements could therefore be estimated (expt 2): maximal weight gain and maximal liver biotin concentration, 0.08 mg/kg; maximal activity of hepatic pyruvate carboxylase and acetyl CoA carboxylase, 0.05 mg/kg; and maximal white muscle pyruvate carboxylase activity, 0.14 mg/kg. No differences were found between fish fed the supplemented nonpurified diet and fish fed its unsupplemented counterpart (expt 3). The biotin requirement of the trout for growth does not exceed that of other vertebrates. These results also raise a question as to the level of supplementation which may be necessary for trout diets under field conditions. PMID- 2563286 TI - The dopamine-1 agonist, SKF 82526, stimulates phospholipase-C activity independent of adenylate cyclase. AB - Dopamine-1 (DA-1) receptors have been found in renal tubular membranes which stimulate both adenylate cyclase and phospholipase-C activity. In renal cortical plasma membrane preparations the DA-1 agonist SKF 82526, forskolin and NaF stimulated adenylate cyclase activity. 2',5'-dideoxyadenosine inhibited basal and DA-1 agonist stimulated adenylate cyclase activity. Forskolin, NaF, dibutyryl cyclic AMP and 2',5'-dideoxyadenosine had no effect on basal or DA-1 agonist stimulated phospholipase-C activity in these membranes. These studies indicate that DA-1 agonist stimulates adenylate cyclase and phospholipase-C activities independently. Phospholipase-C activity was also increased by the nonhydrolyzable GTP analog, guanosine-5'-O-(3-thiophosphate). When DA-1 agonist and guanosine-5' O-(3-thiophosphate) were added together there was a slight but significant increase in phospholipase-C activity. This increase was inhibited in the presence of guanosine-5'-O-(2-thiodiphosphate). DA-1 stimulated phospholipase-C activity was found to be insensitive to both cholera and pertussis toxins. The present studies indicate a cyclic AMP independent transduction pathway for DA-1 receptor mediated through a guanine nucleotide regulatory protein associated phospholipase C. PMID- 2563287 TI - Demonstration of specific dopamine-1 receptor-mediated coronary vasodilation in the anesthetized dog. AB - This study was conducted to identify the vascular dopamine receptor subtype responsible for specific dopamine-mediated coronary vasodilation. The left circumflex coronary artery (LCX) of pentobarbital anesthetized, open chest dogs was isolated and perfused under either constant flow or constant pressure conditions with blood withdrawn from a cannulated left femoral artery. In animals subjected to constant flow LCX perfusion, after beta adrenoceptor blockade with nadolol (4 mg/kg), alpha-1 adrenoceptor blockade with prazosin (0.5 mg/kg) and alpha-2 adrenoceptor blockade with idazoxan (1.0 mg/kg), intracoronary (c) injection of dopamine (0.01-10 micrograms/kg) produced a dose-dependent decrease in LCX perfusion pressure which was unaltered by administration of the dopamine-2 receptor antagonist domperidone (10 micrograms/kg) but which was blocked completely by the dopamine-1 receptor antagonist SK&F R-83566 (5 micrograms/kg). Similarly, under conditions of constant pressure LCX perfusion and after combined beta, alpha-1 and alpha-2 adrenoceptor blockade, i.c. administration of dopamine produced a dose-related increase in LCX coronary blood flow which was inhibited by SK&F R-83566 but not by domperidone. Direct i.c. administration of the selective dopamine-1 receptor agonist, fenoldopam (1 microgram/kg), resulted in an increase in LCX coronary blood flow which was eliminated completely after administration of SK&F R-83566. Doses of the selective dopamine-2 receptor agonist, dipropyldopamine (0.1-30 micrograms/kg), effective in producing blood flow increases in the femoral vascular bed and which could be antagonized by domperidone, produced only minimal and inconsistent changes in LCX coronary blood flow. Our data demonstrate that direct, dopamine-mediated coronary vasodilation in vivo occurs via stimulation of a vascular receptor of the dopamine-1 subtype. PMID- 2563288 TI - ICI 147,798: a slowly dissociable beta adrenoceptor antagonist that causes insurmountable beta-1 and surmountable beta-2 adrenoceptor antagonism in isolated tissues. AB - ICI 147,798 has been shown to exhibit both diuretic and beta-antagonist properties in vivo. The present study investigated the nature and selectivity of the beta-antagonism in a variety of isolated tissues. ICI 147,798 produced a concentration-dependent suppression of the maximum chronotropic response of norepinephrine in guinea pig right atria (beta-1 adrenoceptor). ICI 147,798 caused a concentration-dependent shift to the right of the salbutamol concentration-response curve in the guinea pig trachea (beta-2 adrenoceptor), and Schild analysis suggested competitive inhibition. Propranolol produced parallel shifts to the right of the norepinephrine concentration-response curve in guinea pig right atria, except at relatively high concentrations. The inhibitory effects of propranolol in guinea pig right atria were reversed by greater than 95%, whereas the effects of ICI 147,798 were only slightly reversed after a 6-hr washout period. Preincubation of propranolol with ICI 147,798 in guinea pig right atria prevented completely the suppression of the norepinephrine maximum chronotropic response. Postincubation of propranolol with ICI 147,798 partially reversed the suppression of the maximum chronotropic response. ICI 147,798 had no effect on the maximum chronotropic responses of either histamine (H2-receptor) or forskolin (adenylate cyclase activation) in guinea pig right atria and had no effect on agonist responses in a variety of other receptor systems. The insurmountable beta-1 adrenoceptor antagonism was evaluated based on the assumptions of irreversible competitive antagonism, mixed competitive and noncompetitive antagonism and slowly dissociating competitive antagonism ("hemi equilibrium" conditions). Concentration-dependent changes in norepinephrine KA values suggested the first three possibilities were unlikely.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563289 TI - Insurmountable beta receptor blockade by ICI 147,798 in rabbits. AB - In rabbits, the characteristics of cardiac beta-1 receptor blockade produced by ICI 147,798, a novel beta receptor blocking agent with diuretic properties, were evaluated and compared with those of propranolol. In conscious rabbits, i.v. injections of 0.31, 1.0 and 3.1 mg/kg of ICI 147,798 and 1.0 mg/kg of propranolol caused significant bradycardia. ICI 147,798 produced a dose-dependent shift to the right of the dose-response (chronotropic) curve of isoproterenol with suppression of the maximal tachycardia, an effect characteristic of insurmountable beta receptor blockade. Propranolol also produced a shift to the right of the dose-response curve of isoproterenol without affecting the maximal tachycardia. ICI 147,798-induced antagonism was specific for beta adrenoceptors as it failed to modify the effects of acetylcholine, angiotensin II, phenylephrine, adenosine, histamine and prostaglandin E2 on mean arterial pressure and heart rate. In rabbits with prior autonomic blockade, ICI 147,798, like propranolol, failed to inhibit the positive chronotropic effects of theophylline which are mediated by postreceptor mechanisms. In reserpinized rabbits, ICI 147,798 was found to have no intrinsic sympathomimetic activity. Unlike the effects of propranolol, which were attenuated by first-pass through the hepatic vascular bed, the effects of ICI 147,798 were unaffected suggesting an absence of first-pass metabolism. The effects of propranolol (1.0 mg/kg i.v.) were not detectable at 24 hr after injection, whereas significant beta receptor blocking activity was still present at 24 hr after ICI 147,798 (1.0 mg/kg i.v.). The results suggest that ICI 147,798 is a specific, long-acting, insurmountable beta-1 receptor blocking agent without intrinsic sympathomimetic activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563290 TI - Inhibition of naloxone-resistant antinociception by centrally administered H2 antagonists. AB - In order to test directly the hypothesis that brain histamine and H2-receptors mediate the naloxone-resistant footshock-induced antinociception (FSIA) elicited by 3 min of 3.5 mA, the effect of i.c.v. H2-antagonists on this response was determined. Intracerebroventricular zolantidine dimaleate produced an inhibition of the response that was both time- and dose-dependent, with no effect on base line responses. A dose of 20 micrograms produced maximal (93%) inhibition of FSIA when given 10 min before footshock, with a calculated ID50 of 7.18 nmol. Eight additional H2-antagonists with varying structures and brain-penetrating qualities also exhibited dose-dependent inhibition of the response with no effect on base line nociception. The potencies of these compounds for inhibition of naloxone resistant FSIA correlated significantly (r = 0.966, P less than .001; rho = 0.980, P less than .002) with their affinities at the H2-receptor, providing strong support for the hypothesis that brain histamine and H2-receptors are important components of naloxone-resistant FSIA. PMID- 2563291 TI - Discriminative stimulus properties of midazolam in the pigeon. AB - Five pigeons were trained to discriminate injections of midazolam (1.0 or 3.0 mg/kg i.m.) from saline with responding maintained under a fixed-ratio 30 schedule of food delivery. When other benzodiazepines were tested, they consistently produced greater than 80% of midazolam-appropriate responding. The order of potency for substituting for midazolam was triazolam greater than alprazolam = diazepam = lorazepam greater than midazolam greater than flurazepam = nitrazepam greater than nordiazepam. The barbiturate phenobarbital (10-100 mg/kg) substituted for midazolam in three of four pigeons whereas pentobarbital (1.0-30 mg/kg) substituted in only two of five pigeons. Several nonbenzodiazepine anxiolytics were also evaluated. Methaqualone (3.0-56 mg/kg) substituted in only one of four pigeons and meprobamate (30-100 mg/kg) failed to substitute in any pigeon tested. CL 218,872, when administered either i.m. (0.3-30 mg/kg) or p.o. (1.0-56 mg/kg), and buspirone (0.3-30 mg/kg) did not substitute for midazolam. Compounds from pharmacological classes not related to midazolam also failed to substitute for midazolam. Pretreatment with the benzodiazepine antagonist flumazenil (Ro 15-1788; 0.03-1.0 mg/kg) antagonized the discriminative stimulus properties of midazolam in a dose-related manner in all pigeons tested. However, this antagonism could not be overcome with increasing doses of midazolam in all pigeons. The results of the present study demonstrate that midazolam is an effective discriminative stimulus in the pigeon. Benzodiazepine anxiolytics, but not other compounds with sedative and/or anxiolytic properties, were found to reliably substitute for midazolam. These results suggest that the discriminative stimulus effects of midazolam are pharmacologically selective. PMID- 2563292 TI - Neuronal uptake and metabolism of 2- and 6-fluorodopamine: false neurotransmitters for positron emission tomographic imaging of sympathetically innervated tissues. AB - The neuronal uptake and metabolism of 2-fluorodopamine (2F-dopamine), 6 fluorodopamine (6F-dopamine) and tritium-labeled dopamine were compared in heart, submaxillary gland and spleen of rats to assess the utility of 18F-labeled 2F- or 6F-dopamine for positron emission tomographic imaging of sympathetically innervated tissues. Tritiated dopamine with and without 2F- or 6F-dopamine, or tritiated 2F-dopamine alone, were injected i.v. into rats that were or were not pretreated with desipramine to block catecholamine neuronal uptake or with reserpine to block vesicular translocation of catecholamines. Tissue and plasma samples were obtained at intervals up to 1 hr after injections. At 1 hr after injection of tritiated dopamine, tritium-labeled norepinephrine, dopamine, dihydroxyphenylacetic acid and dihydroxyphenylglucol accounted for less than 2% of the tritium in plasma but up to 92% of that in tissues; tritiated norepinephrine accounted for 70% or more of the tritium in tissues. In contrast, at 1 hr after injection of tritiated 2F-dopamine, tritiated 2F-norepinephrine accounted for 30 to 46% of the tritium in tissues. Desipramine and reserpine pretreatment blocked the tissue accumulation of tritiated and fluorinated dopamine as well as their dihydroxy-metabolites, indicating that accumulation of exogenous norepinephrine and dopamine analogs was within sympathetic storage vesicles. Relative to the doses of dopamine precursors, less 2F- and 6F norepinephrine accumulated in tissues than tritiated norepinephrine, due largely to inefficient beta-hydroxylation of fluorinated dopamine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563293 TI - [3H]-[H-D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2] ([3H]CTOP), a potent and highly selective peptide for mu opioid receptors in rat brain. AB - The cyclic, conformationally restricted octapeptide [3H]-[H-D-Phe-Cys-Tyr-D-Trp Orn-Thr-Pen-Thr-NH2] ([3H]CTOP) was synthesized and its binding to mu opioid receptors was characterized in rat brain membrane preparations. Association rates (k+1) of 1.25 x 10(8) M-1 min-1 and 2.49 x 10(8) M-1 min-1 at 25 and 37 degrees C, respectively, were obtained, whereas dissociation rates (k-1) at the same temperatures were 1.93 x 10(-2) min-1 and 1.03 x 10(-1) min-1 at 25 and 37 degrees C, respectively. Saturation isotherms of [3H]CTOP binding to rat brain membranes gave apparent Kd values of 0.16 and 0.41 nM at 25 and 37 degrees C, respectively. Maximal number of binding sites in rat brain membranes were found to be 94 and 81 fmol/mg of protein at 25 and 37 degrees C, respectively. [3H]CTOP binding over a concentration range of 0.1 to 10 nM was best fit by a one site model consistent with binding to a single site. The general effect of different metal ions and guanyl-5'-yl-imidodiphosphate on [3H]CTOP binding was to reduce its affinity. High concentrations (100 mM) of sodium also produced a reduction of the apparent mu receptor density. Utilizing the delta opioid receptor specific peptide [3H]-[D-Pen2,D-Pen5]enkephalin, CTOP appeared to be about 2000-fold more specific for mu vs. delta opioid receptor than naloxone. Specific [3H]CTOP binding was inhibited by a large number of opioid or opiate ligands.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563294 TI - CGS 20625, a novel pyrazolopyridine anxiolytic. AB - CGS 20625 (2-(4-methoxyphenyl)2,3,5,6,7,8,9,10-octa hydrocyclohepta[b]pyrazolo [3,4-d]pyridin-3-one) is a potent and selective ligand for the central benzodiazepine receptor (IC50 = 1.3 nM), with little or no affinity to several other neurotransmitter receptor binding sites in vitro. CGS 20625 had a gamma aminobutyric acid ratio of 0.9 and increased t-[35S]butylbicyclophosphorothionate binding by 20% in vitro, a profile indicative of a partial agonist or mixed agonist/antagonist. In vivo, CGS 20625 blocked a pentylenetetrazol discriminative cue with an ED50 = 1.7 mg/kg p.o. The compound selectively increased conflict responding in the Cook-Davidson paradigm with a minimal effective dose of 0.3 mg/kg p.o., as compared with 3.0 mg/kg p.o. for diazepam. At doses as high as 100 mg/kg p.o., CGS 20625 had no effect on variable interval responding, suggesting minimal sedation. Unlike diazepam, CGS 20625 had no effect on rotorod performance at doses up to 100 mg/kg p.o. indicating no overt muscle relaxation, and did not potentiate the action of ethanol in this behavioral paradigm. Also, CGS 20625 had no marked effect on locomotor behavior, did not potentiate hexobarbital sleep time and had no sedative activity at doses up to 300 mg/kg p.o. CGS 20625 was efficacious in preventing pentylenetetrazol-induced seizures (ED50 = 0.7 mg/kg p.o.), had less efficacy with no clear dose-response relationship against picrotoxin-induced seizures and had no effect on either strychnine or electroshock-induced convulsions at doses up to 300 mg/kg p.o.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563295 TI - Absence of spare autoreceptors regulating dopamine agonist inhibition of tyrosine hydroxylation in slices of rat striatum. AB - Incubation of rat striatal slices with forskolin (0.05-10 microM) elicited a dose dependent increase in the activity of tyrosine hydroxylase (TH) assayed in subsequently solubilized extracts of the enzyme. At low concentrations (33 microM) of the cofactor (6R)-5,6,7,8-tetrahydro-L-biopterin dihydrochloride TH activity was increased 2.5 to 3-fold. Kinetic analysis of TH activity as a function of (6R)-5,6,7,8-tetrahydro-L-biopterin dihydrochloride concentration indicated that the enzyme isolated from control slices was composed of multiple species with different Km's for cofactor. Treatment with forskolin (1.5-15 microM) converted the enzyme into a single species with a low Km (28 microM) for (6R)-5,6,7,8-tetrahydro-L-biopterin dihydrochloride. The dopamine (DA) agonist R (-)-N-n-propylnorapomorphine (0.1 microM) reversed forskolin-induced activation of TH. Concentration-response curves were obtained for inhibition of forskolin stimulated TH by R-(-)-N-n-propylnorapomorphine and the DA autoreceptor-selective agonists (+)- and (-)-3-(3-hydroxyphenyl)-N-n-propylpiperidine and 3-[4-(4-phenyl 1,2,3,6-tetrahydropyridyl-1)-butyl]indole. R-(-)-N-n-propylnorapomorphine maximally inhibited forskolin-stimulated activity 85%, as indicated by ALLFIT computer analysis of concentration-response curves. (+)-3-(3-hydroxyphenyl)-N-n propylpiperidine and 3-[4-(4-phenyl-1,2,3,6-tetrahydropyridyl-1)-butyl]indole produced a lower degree of maximal inhibition (54 and 63%, respectively), whereas (-)-3-(3-hydroxyphenyl)-N-n-propylpiperidine was inactive. The D2 DA receptor blocker sulpiride (1 microM) competitively antagonised the effects of all the agonists.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563296 TI - A new series of tricyclic (aryloximino)propanolamines displaying very high selective beta 2-blocking properties. AB - A new class of indanones 4 easily obtained by aryne type condensations, followed by transposition of the benzocyclobutanols 3 thus formed, were transformed into the corresponding oximinopropanolamines 7. These compounds were studied for their potential beta-blocking properties. It was found that 7 have generally low beta 1 blocking properties. Their beta 2-blocking action varies from low to very high. Interestingly one of them (7b) has the highest beta 2 activity/beta 1 activity (343) value known to date. PMID- 2563297 TI - First clinical experience with a Q-switched neodymium:YAG laser for urinary calculi. AB - Animal studies using a high intensity nanosecond pulsed neodymium:YAG laser did not reveal any serious tissue damage. Following these investigations patient treatment was begun in June 1987. Laser energy of a neodymium:YAG laser with an 8 nsec. pulse duration and a repetition rate of up to 50 Hz. was coupled into a flexible 600 resp. 400 micron. quartz fiber. Laser-induced breakdown was created with 35 to 50 mJ. at the fiber tip, resulting in a shock wave that disintegrated the calculus into tiny fragments. A total of 56 patients with 58 calculi (54 ureteral and 4 kidney stones) was treated from June 1987 to March 1988. Of the calculi 48 could be fragmented completely, while 6 others were reduced to a size small enough to be removed with forceps. Four stones composed of calcium oxalate monohydrate could not be disintegrated. The combination of laser stone disintegration with flexible ureterorenoscopy implies the possibility of an atraumatic, 1-step procedure for fragmentation of ureteral and kidney calculi. PMID- 2563298 TI - Functional and pharmacological effects of ureteral diversion. AB - Urinary diversion is the final therapeutic approach in several bladder pathologies including selected cases of bladder cancer, neurogenic bladder, and painful bladder syndrome. While there have been a few experimental studies on the effects of urinary diversion and subsequent undiversion on bladder capacity, the pharmacological changes occurring with diversion and undiversion have not yet been investigated. Our primary objectives were to determine the functional and pharmacological alterations in the defunctionalized bladder and the effects of refunctionalization on these changes. Thirty-two male adult canines weighing 15 to 20 lb. were used for this study. Three groups of dogs were urine diverted for one, three, and six month durations (four dogs per group). Another three groups of dogs were urine diverted for one, three, and six months (four dogs per group); these dogs were then undiverted for three months. Six control animals received either one or two sham operations at one, three, and six months. In all groups the functional and contractile characteristics of the bladders were assessed by urodynamics and in vitro contractile studies; bladder weight and muscarinic receptor density were also measured. Intravesical capacities determined by in vivo cystometry were reduced significantly to 74%, 63%, and 47% of control values at one, three, and six months respectively after urinary diversion. Bladder capacity returned to above normal levels in bladders diverted and then subsequently undiverted. Similarly, the compliance and bladder weight of the diverted bladders were significantly less than control, while diverted bladders subsequently undiverted were similar to controls. Maximal bladder contractility in response to bethanechol stimulation was less in diverted bladders compared to control. This decrease in contractility was accompanied by a decrease in muscarinic receptor density. After undiversion maximal bladder contractility to bethanechol reached control levels; this was accompanied by a parallel increase in muscarinic receptor density to control values. There was no effect of diversion or undiversion on the maximal response of bladder strips to methoxamine stimulation. Thus, a bladder that has been diverted (defunctionalized) for a period of time showed decreases in capacity, compliance, and contractility to muscarinic stimulation along with a decrease in muscarinic receptor density. All of these parameters were restored after refunctionalization of the bladder. PMID- 2563299 TI - Down-regulation of glucocorticoid receptor and its relationship to the induction of rat liver tyrosine aminotransferase. AB - Administration of polyvinyl alcohol containing hydrocortisone (F-PVA) to rats twice caused a rapid increase of plasma glucocorticoids (GCs) around 30 micrograms/dl for 3 days. This rise led to decrease of glucocorticoid receptor (GR) in rat liver cytosol and spleen cells 1 h post F-PVA and remained at low level for more than 10 days. Two hours after administration of hydrocortisone 5 mg/100 g b. wt, the liver tyrosine aminotransferase (TAT) activity in normal rats increased from 13.5 +/- 2.9 to 50.7 +/- 8.9 units, while in rats injected with F PVA 3 days ago it increased only from 13.9 +/- 2.3 to 21.0 +/- 6.3 U. The inductivity of rat liver remained low at 7, 11 and 20 days after the injection of F-PVA and recovered 30 days later. This study demonstrates the presence of down regulation of GR in intact animal, and shows that the decrease of GR is accompanied by reduction of target organ response to GCs. PMID- 2563300 TI - Excitatory amino acids inhibit stimulated phosphoinositide hydrolysis in the rat prefrontal cortex. AB - In rat prefrontal cortical slices, the excitatory amino acids N-methyl-D aspartate (NMDA), ibotenate, L-aspartate, quisqualate, kainate and L-glutamate inhibit carbachol-induced phosphoinositide hydrolysis as measured by the accumulation of [3H]inositol-1-phosphate ([3H]IP1). NMDA dose-dependently inhibited the carbachol response (IC50 = 14.4 microM), and this inhibition was blocked by the NMDA receptor antagonist D,L-aminophosphonovaleric acid. Lowering medium Na+ concentration to 10 mM or exposing slices to pertussis toxin alleviated the inhibitory effect of NMDA on carbachol-induced [3H]IP1 formation. Serotonin-induced stimulation of [3H]IP1 was also inhibited by NMDA; in contrast, stimulation by norepinephrine, epinephrine or dopamine was unaffected. The results suggest that excitatory amino acids, besides their traditional role as stimulatory substances, can also act to inhibit the production of 2nd messengers activated by certain neurotransmitters in the brain. PMID- 2563301 TI - Regulation of glutamate and GABA transport by adrenoceptors in primary astroglial cell cultures. AB - In astrocytes grown in primary cultures from cerebral cortex of neonatal rats, alpha 1-adrenoceptors regulate the active uptake of glutamate followed by an activation of glutamic oxaloacetate transaminase (GOT; EC 2.6.1.1.) and a slight activation of glutamine synthetase (GS; EC 6.3.1.2.) activity. The beta adrenoceptors regulate the active uptake of GABA, and this is followed by an activation of gamma-aminobutyric acid alpha-ketoglutarate transaminase (GABA-T; EC 2.6.1.19.). The data suggest that astrocyte adrenoceptors may modulate neurotransmitter induced neuronal excitability. PMID- 2563302 TI - Structural features determining activity of phenothiazines and related drugs for inhibition of cell growth and reversal of multidrug resistance. AB - Phenothiazines and structurally related compounds inhibit cellular proliferation and sensitize multidrug-resistant (MDR) cells to chemotherapeutic agents. To identify more potent pharmaceuticals, we studied the structure-activity relationships of 30 phenothiazines and related compounds on cellular proliferation and MDR in sensitive MCF-7 and resistant MCF-7/DOX human breast cancer cells. Substitutions on the phenothiazine ring that increased hydrophobicity increased antiproliferative and anti-MDR activities. For example, Cl and -CF3 groups increased whereas -OH groups decreased potency. Modifying the length of the alkyl bridge and the type of amino side chain also influenced potency. Compounds with increased activity against cellular proliferation and MDR possessed a four-carbon bridge rather than a three- or two-carbon bridge and a piperazinyl amine rather than a noncyclic amino group. Compounds with tertiary amines were better anti-MDR agents than those with secondary or primary amines but were equipotent antiproliferative agents. The effects of these substituents were unrelated to hydrophobicity. The structure-activity relationships suggest that an ideal phenothiazine structure for reversing MDR has a hydrophobic nucleus with a -CF3 ring substitution at position 2, connected by a four-carbon alkyl bridge to a para-methyl-substituted piperazinyl amine. We subsequently studied related compounds having certain of these properties. Substitution of a carbon for a nitrogen at position 10 of the tricyclic ring, with a double bond to the side chain (thioxanthene), further increased activity against MDR. For example, (trans)-flupenthixol, the most potent of these compounds, increased the potency of doxorubicin against MDR cells by 15-fold, as compared with its stereoisomer (cis)-flupenthixol (5-fold) or its phenothiazine homolog fluphenazine (3-fold). (cis)- and (trans)-flupenthixol were equipotent antiproliferative agents. (trans) flupenthixol was not accumulated more than (cis)-flupenthixol in MDR cells, implying that their stereospecific anti-MDR effects were not the result of selective differences in the access of the drugs to intracellular targets. Both drugs increased the accumulation of doxorubicin in MDR cells, but not in sensitive cells, suggesting that they modulate MDR by interacting with a uniquely overexpressed cellular target in these resistant cells. The apparent lack of clinical toxicity of (trans)-flupenthixol makes it an attractive drug for further investigation. PMID- 2563303 TI - Chronic somatostatin treatment induces enhanced forskolin-stimulated cAMP accumulation in wild-type S49 mouse lymphoma cells but not in protein kinase deficient mutants. AB - Many different types of cells exhibit a supersensitivity of adenylate cyclase after chronic treatment with inhibitory drugs; this phenomenon is manifested by enhanced cAMP accumulation upon removal of the inhibitory drug. Acute treatment of wild-type S49 cells with the somatostatin analog SMS 201-995 (SMS) results in inhibition of cAMP accumulation. We have found that chronic SMS treatment of S49 cells results in enhanced isoproterenol- and forskolin-stimulated cAMP accumulation after removal of the SMS. The forskolin-stimulated cAMP synthetic rate was about 57% higher in SMS-pretreated cells (14.22 +/- 1.02 pmol of cAMP/10(6) cells/min) than in untreated control cells (9.08 +/- 0.84 pmol of cAMP/10(6) cells/min). The time course of forskolin-stimulated intracellular cAMP accumulation is complex, with desensitization of cAMP synthesis and marked egress of cAMP from the cells. We have modeled the forskolin-stimulated cAMP time course to a simple function incorporating the initial synthetic rate and rate constants for desensitization and elimination (degradation plus egress). The mathematical modeling suggests that the difference in forskolin-stimulated cAMP time courses between control and SMS-pretreated cells can be explained on the basis of a difference in initial synthetic rates. We tested the hypothesis that the SMS induced change in forskolin-stimulated cAMP accumulation is triggered by the decrement in the concentration of intracellular cAMP caused by SMS. We studied two independently isolated mutants of S49 cells that are devoid of cAMP-dependent protein kinase activity (kin-). Although SMS acutely inhibits cAMP accumulation in both kin- mutants, neither mutant exhibited an enhanced forskolin-stimulated cAMP synthetic rate after chronic SMS treatment. These results suggest that cAMP dependent protein kinase is important in the induction of adenylate cyclase supersensitivity in wild-type S49 cells. The mechanistic signal for induction of supersensitivity may be the decreased cAMP accumulation that occurs in response to stimulation of inhibitory receptors, although other hypothetical mechanisms may be invoked. PMID- 2563304 TI - Differences in dopamine receptor reserve for N-n-propylnorapomorphine enantiomers: single unit recording studies after partial inactivation of receptors by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline. AB - Previous studies with (S)-(+)-N-n-propylnorapomorphine (NPA), an enantiomer of the potent dopaminergic agonist (R)-(-)-NPA, showed that this compound had a complex pharmacological profile. For example, (S)-(+)NPA had weak dopaminergic agonist potency, in that it could slow and ultimately stop firing of substantia nigra and ventral tegmental area dopamine neurons. However, antagonist properties were also evident inasmuch as immediate pretreatment with a low dose of (S)-(+) NPA caused a significant rightward shift of the (R)-(-)-NPA dose-response curve. This dual agonist-antagonist nature of (S)-(+)-NPA suggested a low intrinsic efficacy for (S)-(+)-NPA. To test this hypothesis, we conducted dose-response studies for the inhibition of rat substantia nigra dopamine cell firing by (R)-( )- and (S)-(+)-NPA 1 day after partial irreversible dopamine receptor inactivation with 6 mg/kg N-ethoxycarbonyl-2-ethoxy-1-2-dihydroquinoline (EEDQ) in an ethanol vehicle. This degree of inactivation resulted in a significant, parallel, rightward shift of the (R)-(-)-NPA dose-response curve relative to ethanol-treated control rats, with no loss of maximal response. After pretreatment with the same dose of EEDQ, however, the maximal response to (S)-(+) NPA was reduced by 22% with no shift on the dose axis. The dose-response curves for control and EEDQ-treated groups were subjected to Furchgott analysis to determine per cent response versus fractional occupancy relationships for each drug. A steep hyperbolic relationship was revealed for (R)-(-)-NPA. Fifty per cent and maximal (greater than 95%) inhibitions of cell firing occurred at 3.5% and approximately 32% receptor occupancies, respectively. Hence, for (R)-(-)-NPA there is about a 68% receptor reserve in this model. The same analysis for (S) (+)-NPA yielded an occupancy versus response plot that was more shallow and linear. Half-maximal effects occurred at 66% occupancy and the maximal response (96% inhibition) required 94% occupancy, indicating few spare receptors for (S) (+)-NPA. Because the ratio of fractional occupancies at a given level of response is a measure of relative efficacy, we calculated a relative efficacy of (S)-(+)- to (R)-(-)-NPA of 0.05 (at the 50% response level). This confirms that (S)-(+) NPA indeed has a much lower intrinsic efficacy than the (R)-(-)-antipode and may account for the previously reported antagonist property of the (+)-form under certain treatment conditions. PMID- 2563305 TI - Agonist-promoted sequestration of the beta 2-adrenergic receptor requires regions involved in functional coupling with Gs. AB - The molecular basis for the desensitization of beta 2-adrenergic receptors was investigated by oligonucleotide-directed mutagenesis. beta-Adrenergic receptor mutants containing deletions within the sixth hydrophilic domain that failed to couple to Gs and stimulate adenylyl cyclase did not undergo agonist-mediated sequestration. In contrast, all receptor mutants that displayed Gs coupling were sequestered away from the cell surface in response to isoproterenol. Progressive truncation of the C-terminus of the receptor resulted in decreases in the initial rates of receptor sequestration and functional uncoupling, although the final extent of these desensitization processes was not affected by the mutations. These data suggest that structural features of the beta 2-adrenergic receptor that are involved in receptor activation are also essential for mediating the subsequent inactivation caused by the sequestration of the receptor from the cell surface. PMID- 2563306 TI - Acute in vivo amphetamine produces a homologous desensitization of dopamine receptor-coupled adenylate cyclase activities and decreases agonist binding to the D1 site. AB - We have previously reported that, 30 min after a single injection of 7.5 mg/kg d amphetamine sulfate, there was a significant 25% decrease in the apparent Vmax for stimulation of adenylate cyclase activity by the D1 receptor-selective partial agonist SKF 38393 in rat striatal membranes, as compared with saline injected controls. This desensitization was seen in the striatal but not the mesolimbic forebrain. In the present study this desensitization was further characterized by using various ligands that interact with the three components of the D1 receptor-coupled adenylate cyclase complex to determine the site of modification that resulted in the desensitization. The desensitization was not associated with a change in the stimulation of adenylate cyclase at the level of the catalytic subunit or the guanyl nucleotide-regulatory protein Ns. Receptor number, as assessed by the binding of the D1 selective antagonist [3H]SCH 23390, was unaltered in the desensitized state. In contrast, the number of high affinity binding sites, as measured with the agonist [3H]dopamine was decreased 30% by acute amphetamine exposure. This suggests that the amphetamine-induced desensitization may be the result of an uncoupling of the receptor from Ns. In order to further assess the effects of amphetamine on receptor/Ns coupling, we measured the ability of the guanyl nucleotide guanosine-5'-(beta,gamma imido)triphosphate [Gpp(NH)p] to decrease high affinity [3H]dopamine binding to striatal membranes. The inclusion of 100 microM Gpp(NH)p in the assay decreased the number of receptors in the high affinity state by 40% and 52% in membranes from saline- and amphetamine-pretreated rats, respectively. These results imply that amphetamine treatment does not modify the ability of Gpp(NH)p to decrease high affinity agonist binding. It is possible that amphetamine treatment decreases the number of receptors that can couple to Ns but the remaining receptors can still form a high affinity complex and are sensitive to the effects of Gpp(NH)p. We also report that maximal D2 dopamine receptor-mediated inhibition of forskolin-stimulated adenylate cyclase activity was decreased in striatal membranes prepared from amphetamine-treated rats as compared with saline-injected controls, implying that the D2 pathway was desensitized by amphetamine treatment. Conversely, acute amphetamine injection did not alter the ability of either the adenosine A2 receptor to stimulate or the muscarinic cholinergic receptor to inhibit adenylate cyclase activity in the rat striatum. These results suggest that acute amphetamine treatment produces a dopamine receptor-specific or homologous desensitization. PMID- 2563308 TI - [Sulfasalazine therapy in spondylarthritis ankylopoietica]. AB - In the past few years several reports have been published on the favourable effect of sulphasalazine in ankylosing spondylitis. This induced us to compare sulphasalazine with placebo in a prospective, randomized single-blind trial in 63 patients with active ankylosing spondylitis. After termination of the 24 week course of treatment significant improvement was registered in several clinical parameters both in the group taking the active drug (31 patients) and in that on placebo (32 patients). The advantage of sulphasalazine over placebo only proved to be significant in the duration of articular morning stiffness and in reducing disturbances of sleep. Observations claiming sulphasalazine to act as a remission inducing drug for ankylosing spondylitis have neither been confirmed nor ruled out by our experiences. Accurate appraisal of the effectiveness of sulphasalazine still calls for further extensive placebo-controlled double-blind studies. PMID- 2563309 TI - POU! Goes the homeobox. PMID- 2563307 TI - Induction of CAD gene amplification by restriction endonucleases in V79,B7 Chinese hamster cells. AB - The restriction endonucleases PvuII, BamHI and EcoRI were tested for their ability to induce gene amplification in V79,B7 Chinese hamster cells. The results indicate that treatment with these enzymes efficiently increases the frequency of clones resistant to N-phosphonacetyl-L-aspartate, indicating induction of CAD gene amplification. PMID- 2563310 TI - Circulating auto-antibodies to neutrophil cytoplasmic antigens in vasculitis. A report of 3 cases. AB - Three patients are described who presented with symptoms and signs of a vasculitic illness but in whom a definite diagnosis was not made until the application of a test to detect auto-antibodies to neutrophil cytoplasmic antigens. There has long been a need for a diagnostic marker for the groups of diseases which fall into the broad classification of systemic vasculitides. Diagnosis of the patients as having either Wegener's granulomatosis or microscopic polyarteritis by the finding of this auto-antibody enabled a positive approach to treatment with combinations of immunosuppressive agents. In addition, serial measurement of the auto-antibody titres enabled monitoring of disease activity. PMID- 2563311 TI - Effects of alpha- and beta-adrenergic antagonists on plasma apolipoproteins and forearm blood flow in patients with mild hypertension. AB - To study the mechanisms by which adrenergic antagonists affect blood pressure and plasma lipid levels, the effects of alpha-blockade with prazosin were compared with those of beta-blockade with propranolol in 23 normolipidemic, mildly hypertensive patients. Plasma lipoprotein composition, apolipoproteins, and some of the processes involved in lipid synthesis and clearance from plasma were investigated also. Patients entered an eight-week placebo period during which they were free of all antihypertensive medications. They were then randomly assigned under double-blind conditions to treatment with either prazosin (mean dose, 5 mg per day) or propranolol (mean dose, 133 mg per day) for eight weeks. Doses of both drugs were titrated to achieve either a decrease in diastolic blood pressure of 10 mm Hg or more or a reduction of diastolic blood pressure to less than 85 mm Hg, whichever was lower. Total plasma cholesterol decreased by 9 percent during prazosin treatment and increased by 7 percent during propranolol treatment (p less than 0.005 between treatments). Low-density lipoprotein cholesterol decreased by 12 percent with prazosin and increased by 12 percent with propranolol (p less than 0.005). Apolipoprotein B decreased by 17 percent with prazosin and increased by 15 percent with propranolol (p less than 0.005). There were no significant changes in total high-density lipoprotein cholesterol, its subfractions high-density lipoprotein2 or high-density lipoprotein3, or in apolipoprotein A1 and apolipoprotein A2. Plasma very low-density lipoprotein and low-density lipoprotein triglycerides were not significantly affected by either treatment. Plasma post-heparin lipase activities, which clear triglyceride and high-density lipoprotein cholesterol from plasma, were not altered significantly. Since regional blood flow could affect the clearance of plasma lipoproteins, measurements were taken of forearm blood flow, forearm vascular resistance, and maximal forearm vasodilatory potential during reactive hyperemia. The adrenergic antagonists had no effect on these measurements, nor did they affect cellular cholesterol synthesis as measured by the activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase in blood mononuclear cells. The results of this study demonstrate differing actions between alpha- and beta-adrenergic antagonism. Alpha-blockade produced significantly lower levels of plasma low-density lipoprotein cholesterol and apolipoprotein B than beta-adrenergic antagonism without changes in high-density lipoproteins. PMID- 2563312 TI - Amyl nitrite-induced hemolytic anemia. PMID- 2563313 TI - Topics in clinical pharmacology: buspirone: a new nonbenzodiazepine anxiolytic agent. PMID- 2563314 TI - Use of esmolol during anesthesia to treat tachycardia and hypertension. AB - We evaluated the clinical effectiveness of esmolol, an ultra-short-acting, cardioselective beta-adrenergic receptor blocker, in controlling sinus tachycardia and increased systolic blood pressure occurring perioperatively in 30 ASA physical status II or III patients having elective, non-cardiac surgery. Esmolol 80 mg I.V. bolus (N = 15) or placebo (N = 15) followed by 12 mg/min or placebo were infused in 30 isoflurane-anesthetized patients using a randomized double-blind study design. The bolus plus infusions were given when surgical stimuli caused heart rate to exceed 95 bpm or systolic blood pressure 140 mm Hg. Esmolol significantly decreased heart rate (107 +/- 4, mean +/- SEM to 99 +/- 4, mean +/- SEM bpm) within 45 sec after starting the bolus plus infusion; the placebo had no effect, heart rate being 105 +/- 4 before and 106 +/- 3 bpm after the bolus plus infusion. Patients given esmolol continued to have heart rates significantly lower than patients given placebo injections throughout a six min infusion (Ex., at 5 min 81 +/- 3 vs 91 +/- 4 bpm). The study demonstrated no apparent effect of esmolol on blood pressure but that esmolol is effective in treating perioperative sinus tachycardia. PMID- 2563315 TI - Is the priming principle both effective and safe? AB - This study determined the priming dose of vecuronium (V), pancuronium (P) and atracurium (A) that resulted in the most rapid onset of neuromuscular blockade (NMB) in 150 patients given either V 0.08 mg/kg, P 0.1 mg/kg or A 0.6 mg/kg. Patients were further divided (n = 10 per group) to receive no prime or 5%, 10%, 15% or 20% of the total dose as a prime followed 5-7 minutes later by the remaining (intubating) dose. A further 10 patients received 0.04 mg/kg d tubocurarine followed by 1.5 mg/kg succinylcholine (S). Priming significantly shortened the onset of NMB. The priming doses producing the most rapid onset were 0.012 mg/kg for V, 0.015 mg/kg for P and 0.09 mg/kg for A. The S resulted in significantly greater NMB at 60 sec than any priming dose of A, V or P. There was no difference between the three nondepolarizing neuromuscular blockers in shortening the onset of NMB produced by priming. To evaluate both the effect of the "optimal" priming dose in awake patients and the effect of increasing intubating doses on NMB an additional 40 patients were given V 0.012 mg/kg followed by V 0.08, 0.1, 0.12 or 0.15 mg/kg. Increasing the intubating dose did not improve onset of NMB. The "optimal" priming dose, however, resulted in a high incidence of symptoms of muscle weakness. We conclude that priming shortens the onset of NMB similarly between V, P and A but the priming dose producing the most rapid onset of NMB also results in a high incidence of side effects and therefore the priming principle should be used with caution. PMID- 2563316 TI - Does choice of anesthetic agent significantly affect outcome after coronary artery surgery? AB - A prospective study of 1094 consecutive adult patients undergoing coronary revascularization was undertaken to determine the effect of anesthetic technique on outcome. Patients received one of five primary techniques: high-dose fentanyl (greater than 50 micrograms/kg), moderate-dose fentanyl (less than 50 micrograms/kg), sufentanil (3-8 micrograms/kg), diazepam (0.4-1 mg/kg) with ketamine (3-6 mg/kg) or halothane (0.5-2.5% inspired concentration after thiopental induction). Supplemental inhalation anesthesia (enflurane, halothane, or isoflurane) was used in 60% of cases where the primary technique was intravenous based. Patients in the above anesthetic groupings had similar perioperative demographic and risk classifications. The overall incidence of postoperative myocardial infarction, postoperative low cardiac output state, and in-hospital death were 4.1, 5.6, and 3.1%, respectively. There were no significant differences in the incidence of these occurrences or in the incidence of serious pulmonary, renal, or neurologic morbidity or length of ICU stay among primary anesthetic techniques nor among supplemental inhalation agent groups. Multivariate discriminant analysis of this data suggests that a multitude of factors are significantly more important than anesthetic technique as determinants of outcome after coronary artery surgery. PMID- 2563317 TI - The incidence of myocardial ischemia during anesthesia for coronary artery bypass surgery in patients receiving pancuronium or vecuronium. AB - This study was performed to compare the incidence of prebypass myocardial ischemia in patients receiving fentanyl and enflurane for anesthesia along with either pancuronium or vecuronium. Ninety-eight patients with normal left ventricular function were randomly allocated to receive either pancuronium 0.15 mg.kg-1 or vecuronium 0.15 mg.kg-1 in a double-blind manner after fentanyl 40 micrograms.kg-1 for induction of anesthesia for elective coronary artery bypass grafting (CABG). Premedication included diazepam 0.15 mg.kg-1 po, morphine 0.10 mg.kg-1, and scopolamine 0.005 mg.kg-1 im. Two lead Holter monitor recordings (leads V6 and V9) from the time of arrival in the operating suite to institution of cardiopulmonary bypass were analyzed for ischemia by a cardiologist blinded to the choice of muscle relaxant. Intraoperatively, heart rates greater than 90 beats.min-1 and systolic blood pressure +/- 20% of ward values were treated with propranolol, enflurane, or phenylephrine. Nitroglycerin was infused for ECG signs of ischemia or pulmonary hypertension. After induction of anesthesia the heart rate and cardiac index were consistently decreased in patients receiving vecuronium and also lower in these patients compared with those receiving pancuronium. Thirty-two per cent of patients receiving pancuronium received propranolol for heart rates greater than 90 beats.min-1 versus 7% of those who received vecuronium (P approximately 0.01). Eight patients developed 13 episodes of ischemia after administration of the muscle relaxant: four who received pancuronium (n = 44; 9%) and four receiving vecuronium (n = 54; 7%). Four episodes occurred at induction or tracheal intubation, two in each group. There were four perioperative myocardial infarctions as determined by ECG and CPK-MB levels, two in each group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563318 TI - In vitro inhibition of a polymorphic human liver P-450 isozyme by narcotic analgesics. AB - Genetically determined differences in the ability of individuals to oxidize certain drugs by hepatic P-450 processes has gained increasing attention. The so called debrisoquin hydroxylase, which is defective in approximately 5-10% of whites, is known to be of major importance for the metabolism of several drugs. The possible relation of this isozyme to the metabolism of narcotics eliminated by oxidative biotransformation has not been studied. Several narcotics were screened for in vitro interaction with this P-450 isozyme. This was accomplished by testing for competitive inhibition by narcotics of the 2-hydroxylation of desmethylimipramine in a human liver microsomal preparation. Alfentanil, fentanyl, and dextropropoxyphene were found to competitively inhibit this pathway demonstrating an interaction with this polymorphic isozyme. No interaction was found for codeine, meperidine, methadone, morphine, or nalbuphine. These results suggest that a genetic defect may be important for elimination clearance by metabolism for dextropropoxyphene, alfentanil, and fentanyl and that in vivo investigation is warranted. PMID- 2563319 TI - Effect of antiasthma drugs on microvascular leakage in guinea pig airways. AB - We have studied the effect of intravenous epinephrine, albuterol, verapamil, and aminophylline on airway microvascular leakage in guinea pigs. Microvascular leakage was induced by platelet-activating factor (PAF; 50 ng/kg intravenously), which acts directly on venular endothelial cells, and measured by quantifying extravasation of Evans blue (EB) dye. Epinephrine (20 micrograms/kg) inhibited PAF-induced changes in dye leakage in larynx and main bronchi; at 80 and 160 micrograms/kg, significant inhibition was observed in all airways studied. This effect was reversed by phentolamine (2.5 mg/kg) or prazosin (100 micrograms/kg). By contrast, albuterol (20 to 320 micrograms/kg) and aminophylline (12.5 to 50 mg/kg) failed to inhibit dye leakage at any dose studied. Verapamil inhibited PAF increased leakage in larynx, main bronchi, and intrapulmonary airways at the lowest dose tested (125 micrograms/kg), although inhibition was not dose dependent. These results suggest that the antiedema effect of epinephrine may be due to vasoconstriction rather than to a direct effect on endothelial cell contractility and that neither beta-agonists nor theophylline have an inhibitory effect. The inhibitory effect of epinephrine on airway microvascular leakage may have therapeutic implications for asthma. PMID- 2563320 TI - The effect of oral terfenadine alone and in combination with flurbiprofen on the bronchoconstrictor response to inhaled adenosine 5'-monophosphate in nonatopic asthma. AB - Inhaled adenosine and adenosine 5'-monophosphate (AMP) cause bronchoconstriction in atopic and nonatopic asthmatics by a mechanism believed to involve histamine release from airway mast cells and an interaction with neural reflexes. In the present study, we have investigated the effect of oral terfenadine 180 mg, flurbiprofen 100 mg, and the drug combination on AMP-induced bronchoconstriction in eight nonatopic asthmatic subjects with a mean age of 53.8 +/- 5.6 yr. The provocation concentrations of histamine and AMP required to produce a 20% decrease in FEV1 (PC20) were determined to be 2.5 (range, 0.2 to 16.3) and 50.1 (range, 1.5 to 841) mg/ml, respectively, representing a potency difference of 17.8-fold on a molar basis. In subsequent time-course studies, the bronchoconstrictor response to inhalation of the PC20 histamine was suppressed completely by terfenadine and the drug combination, but unaffected by flurbiprofen. Terfenadine alone and flurbiprofen alone inhibited bronchoconstriction provoked by the PC20 AMP by 49.8 +/- 5.5% (p less than 0.01) and 31.9 +/- 7.9% (p less than 0.01), respectively, when areas under the FEV1 time curves were compared with placebo, the difference between the two treatments not being significant (p = 0.06). The drug combination inhibited the response to AMP by 60.0 +/- 8.3% (p less than 0.01), this being significantly greater than with flurbiprofen (p less than 0.01), but not with terfenadine alone. These data implicate both histamine and cyclooxygenase products in the bronchoconstrictor response to AMP. PMID- 2563321 TI - The effect of six months of daily treatment with the beta-2 agonist oral pirbuterol on pulmonary hemodynamics in patients with chronic hypoxic cor pulmonale receiving long-term oxygen therapy. AB - We studied the acute and long-term effect of the oral beta-2 agonist pirbuterol on pulmonary hemodynamics and right and left ventricular ejection fractions in 11 patients with stable hypoxic chronic bronchitis and emphysema who were maintained on long-term oxygen therapy. We have used a double-blind study with random allocation of patients to receive either pirbuterol or an identical-appearing placebo. Six patients received 15 mg pirbuterol thrice daily, and five patients received a similar placebo thrice daily, over a 6-month period. The first 15-mg dose of pirbuterol by mouth at the beginning of the study raised the heart rate from 84 +/- 13 to 100 +/- 16 beats/min (mean +/- SD) (p less than 0.01), and cardiac output from 5.2 +/- 0.9 to 6.4 +/- 0.8 L/min (p less than 0.01) and right ventricular ejection fraction (RVEF) from 0.28 +/- 0.10 to 0.33 +/- 0.12 (p less than 0.01) within 120 min after receiving the drug, but without significant change in the mean pulmonary arterial pressure. None of these variables changed significantly after an acute dose of placebo in another four patients. Repeated measurements after 6 months of chronic oral therapy with either pirbuterol or placebo showed that 16 h after the last 15-mg oral dose of pirbuterol or placebo, the heart rate, mean pulmonary arterial pressure, cardiac output, total pulmonary vascular resistance, and RVEF were all not significantly different from the values 6 months previously, before receiving either pirbuterol or the placebo.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563322 TI - Identification and characterization of a homogeneous population of beta 2 adrenergic receptors on human alveolar macrophages. AB - Human alveolar macrophages obtained by bronchoalveolar lavage were studied with the high specific activity beta-adrenergic ligand [125I]pindolol and found to possess a moderate density of beta-adrenergic receptors. Using macrophage membranes, the receptor density (Bmax) was 42 +/- 9 fmol/mg protein with an apparent equilibrium dissociation constant (Kd) of 44 +/- 9 pM (mean +/- SEM). With intact macrophages, the Bmax = 5,643 +/- 942 sites/cell with Kd = 29 +/- 9 pM. Competition binding studies with subtype-specific antagonists revealed an exclusive population of beta 2-adrenergic receptors. Incubation of intact macrophages with the beta-adrenergic agonist isoproterenol caused a 6-fold increase in intracellular cyclic AMP (cAMP). Prostaglandin E1 and forskolin, which activate adenylate cyclase via different mechanisms, afforded typical marked increases in macrophage cAMP. Saturation binding, competition binding, and cAMP accumulation studies may all be performed from a single sample of about 2 x 10(7) cells, which can be obtained by bronchoalveolar lavage. This should facilitate studies of in vivo regulation of human alveolar macrophage beta adrenergic receptors with regard to immune function and mediator release, and as a possible reflection of lung parenchymal receptors. PMID- 2563323 TI - Hemorrhagic fever with renal syndrome: four decades of research. AB - Hemorrhagic fever with renal syndrome is an acute infectious illness characterized by fever, hemorrhage, and renal failure. Research over the last 40 years has led to the discovery and characterization of the causative viruses, detailed knowledge of the epidemiology of the disease, development of sensitive diagnostic assays, and improvements in patient management, which in turn have led to significant reductions in mortality. Considerable progress has also been made in elucidating the pathophysiology of the disease, although much more needs to be learned. Recent data show that hemorrhagic fever with renal syndrome has a wider geographic occurrence than previously thought. This syndrome must be considered in the differential diagnosis of acute renal failure of unknown cause. PMID- 2563324 TI - Hemorrhagic fever with renal syndrome and Tamm-Horsfall protein. PMID- 2563325 TI - Mosquito salivary gland antigens identified by circulating human antibodies. AB - Salivary glands were removed from female mosquitoes of laboratory-reared strains of species common to southern Florida. Protein antigens were isolated from homogenates of these glands by denaturing electrophoresis and transferred to nitrocellulose for immunoblotting with serum samples obtained from human volunteers. A spectrum of antigens with a wide range of molecular weights (14 to 126 kilodaltons) were identified by antibodies in human serum for each species. Both species-unique and species-shared antigens were present. The results of these studies indicate that the humoral response to mosquito bite is complex, with a multitude of antigens provoking antibody responses. Since each individual has his or her own unique exposure history to mosquitoes, it seems unlikely that desensitization to a specific immunogen will confer protection in clinical situations in which multiple exposures to a variety of mosquito species occur. PMID- 2563326 TI - 'Ecstasy': a human neurotoxin? PMID- 2563327 TI - Home-based treatment of obsessive-compulsive patients: intersession interval and therapist involvement. AB - In a 2 x 2 factorial design massed vs spaced and therapist vs self-controlled exposure were compared with obsessive-compulsive patients. Intersession interval was varied keeping constant the length of exposure time and number of exposure sessions. Treatment in all conditions was home based. Treatment led to highly statistical and significant improvements on all measures. Massed exposure was as effective as spaced exposure; self-controlled exposure proved to be as effective as therapist-controlled exposure. PMID- 2563328 TI - Kinetic properties, nutrient-dependent regulation and energy coupling of amino acid transport systems in Penicillium cyclopium. AB - In submerged grown hyphae of Penicillium cyclopium the activities of seven transport systems could be distinguished which share in the uptake of L-arginine, L-glutamic acid, L-phenylalanine and L-leucine. They include the specific systems a (accepting L-arginine and L-lysine), b (L-phenylalanine, L-tyrosine), c (L glutamic acid) and d (L-leucine), system I (a 'general amino-acid permease') and the low-affinity systems II and III, which accept acidic or basic amino acids, respectively, but also L-phenylalanine. In nutrient-sufficient cells, systems I, II and III remain repressed; uptake is dominated by the specific systems b, c, d and a, the latter reaching its maximum activity. Nitrogen starvation is the most powerful signal for the development of systems I, II and III, whereas, in carbon starved cells, systems b, c and d reach maximum activities. The development of the general amino-acid permease in nitrogen-starved cells requires both translational and--with a few hours delay--transcriptional events as indicated by the influence of cycloheximide and 5-fluorouracil. The uptake of all amino acids is accompanied by a transient acidification of the cellular interior. Short-time preaccumulation of several anions, such as citrate, alpha-oxo-glutarate, glutamate (but not glutamine), increases the initial rate of amino-acid uptake at a pH above the optimum. Uncouplers inhibit the uptake not only under aerobic but also under anaerobic conditions, where the ATP content is not influenced by these compounds. These findings point to an H+/amino acid symport, which is tightly connected with the recycling of the incoming protons by the plasmalemma H+ ATPase. PMID- 2563329 TI - Two types of K+ channels in the apical membrane of rabbit proximal tubule in primary culture. AB - The patch-clamp technique was used to investigate ionic channels in the apical membrane of rabbit proximal tubule cells in primary culture. Cell-attached recordings revealed the presence of a highly selective K+ channel with a conductance of 130 pS. The channel activity was increased with membrane depolarization. Experiments performed on excised patches showed that the channel activity depended on the free Ca2+ concentration on the cytoplasmic face of the membrane and that decreasing the cytoplasmic pH from 7.2 to 6.0 also decreased the channel activity. In symmetrical 140 mM KCl solutions the channel conductance was 200 pS. The channel was blocked by barium, tetraethylammonium and Leiurus quinquestriatus scorpion venom (from which charybdotoxin is extracted) when applied to the extracellular face of the channel. Barium and quinidine also blocked the channel when applied to the cytoplasmic face of the membrane. Another K+ channel with a conductance of 42 pS in symmetrical KCl solutions was also observed in excised patches. The channel was blocked by barium and apamin, but not by tetraethylammonium applied to the extracellular face of the membrane. Using the whole-cell recording configuration we determined a K+ conductance of 4.96 nS per cell that was blocked by 65% when 10 mM tetraethylammonium was applied to the bathing medium. PMID- 2563330 TI - [Effects of psychotropic drugs of the phenothiazine series on matrix activity of thymocyte DNA and glucocorticoid receptor interactions]. AB - The experiment on adrenalectomized rats has shown that the concentration rates of 3.5.10(-4)M aminazine and 7.0.10(-4)M tizercine reduce 3H-acetonide triamcinolone receptor interaction, and the concentration rate of 7.0.10(-4) tizercine increases 3H-cortisol receptor interaction in liver cytosol. The concentration rates of 3.5.10(-7)M aminazine and 3.5.10(-5)M tizercine suppress 3H-uridine inclusion into the acid-resisting mRNA thymocyte fraction. Associated introduction of aminazine and tizercine with acetonide triamcinolone (10(-8)M) increases the inhibiting effect on 3H-uridine inclusion into mRNA thymocytes. This suggests that aminazine and tizercine suppress matrix activity of DNA thymocytes through their effect on glucocorticoid receptors of type II. PMID- 2563331 TI - Comparison of the antinociceptive effect of morphine and glutamate at coincidental sites in the periaqueductal gray and medial medulla in rats. AB - In rats, microinjection of the opioid agonist morphine (15-45 nmol/0.5 microliter) and the excitatory amino acid monosodium glutamate (30-60 nmol/0.5 microliter) into identical brainstem sites within the mesencephalic periaqueductal gray matter (PAG; 23 sites) and the rostral ventromedial medulla (RVM; 22 sites) produced an increase of tail flick and hot plate response latencies. At both PAG and RVM sites, there was a statistically significant relationship between the effect obtained with morphine and with glutamate on the two nociceptive responses. While morphine and glutamate produced indistinguishable inhibition of tail flick and hot plate response latencies in the PAG, the tail flick inhibition following RVM morphine, but not RVM glutamate, displayed a clear plateau. One parsimonious interpretation of these data is that (1) glutamate directly increases the activity in the bulbospinal pathway, and (2) morphine inhibits an evoked or tonic suppression of a bulbospinal projection. PMID- 2563332 TI - Long-lasting physiological effects of bath applied N-methyl-D-aspartate. AB - The present experiments describe a long-lasting form of potentiation induced in field CA1 of rat hippocampal slices by bath application of N-methyl-D-aspartate (NMDA), in association with low magnesium concentrations, glycine and spermine. The potentiation effect consisted of a 50% increase in slope of field potentials and was stable for at least 80 min post treatment. It was not accompanied by detectable changes in antidromic responses and was completely blocked by an antagonist of NMDA receptor. The possible relationship of pharmacologically induced potentiation to long-term potentiation (LTP) is discussed. PMID- 2563333 TI - Glutaminase inhibition and the release of neurotransmitter glutamate from synaptosomes. AB - Cerebrocortical synaptosomes were incubated with glutamine together with 6-diazo 5-oxo-L-norleucine (DON) and NH4Cl (which are known to inhibit phosphate stimulated glutaminase) in order to assess the effect of such inhibition on the pool sizes and extent of evoked release of endogenous amino acids, particularly glutamate. DON (5 mM) inhibited glutaminase by 73-89% and NH4Cl (1-4 mM) inhibited the enzyme by 45-53% under the conditions employed. NH4Cl (4 mM) incubated with synaptosomes for 30 min reduced pool sizes of aspartate and glutamate by 28% and inhibited release of glutamate by 55% compared to control release. DON caused a decrease in both the pool size of glutamate (22%) and the extent of veratrine-evoked release of amino acids (21%). PMID- 2563334 TI - Localization and characterization of somatostatin binding sites in the mouse retina. AB - We studied the binding of [125I]Tyr11-somatostatin-14 and [125I]Leu8,D Trp22,Tyr25-somatostatin-28 to frozen, unfixed sections of C57BL/6J mouse eyes with autoradiography. Specific binding of both ligands occurred in 3 maxima, a broad band extending from the retinal ganglion cell to the inner nuclear layers, a narrow and inconstant band over the outer plexiform layer, and a band over the retinal pigment epithelium and choroid. We quantified the label over the inner plexiform layer and found evidence for a single, saturable binding site after Scatchard analysis of saturation binding data. With [125I]Tyr11-somatostatin-14 the dissociation constant (Kd) was 1.48 nM and the total number of binding sites (Bmax) was 68 fmol/mg protein; in competition experiments the inhibitory binding constant (Ki) was 900 pM for somatostatin-14 and 350 pM for somatostatin-28. With [125I]Leu8,D-Trp22,Tyr25-somatostatin-28, Kd was 625 pM and Bmax was 69 fmol/mg protein; in competition experiments Ki was 4.58 nM for somatostatin-14 and 710 pM for somatostatin-28. These results demonstrate the existence of somatostatin receptors in the inner plexiform layer of the retina that appear to have greater specificity for somatostatin-28 than for somatostatin-14. PMID- 2563335 TI - Tryptamine receptors: neurochemical and electrophysiological evidence for postsynaptic and functional binding sites. AB - [3H]Tryptamine binding characteristics and responsiveness of spontaneously active caudate nucleus neurons to intravenous application of drugs were assessed 6 weeks following unilateral application of 6-hydroxydopamine (6-OHDA, 8 micrograms) to the substantia nigra of male Wistar rats. The effects of this lesion procedure on caudatal levels of dopamine, 5-hydroxytryptamine (5-HT) and their acid metabolites, and on pargyline-induced (200 mg/kg, 2 h, s.c.) accumulation of tryptamine in the caudate nucleus were also assessed. Levels of caudatal dopamine and metabolites were reduced ipsilateral to the lesion. Concurrently there was a reduction in the extent of pargyline-induced tryptamine accumulation. Caudatal [3H]tryptamine binding was increased ipsilateral to the lesion, indicating a postsynaptic localization of this binding site. Bmax values in the caudatal samples ipsilateral to the lesion were increased by an average of 34% relative to the contralateral side. Contralateral Bmax values were equivalent to those routinely obtained in control animals. The affinity (Kd) of these binding sites for [3H]tryptamine was unchanged by the lesion procedure. The firing rate of caudate neurons was inhibited by intravenous application of tryptamine, apomorphine and 5-MeODMT. The lesion procedure did not affect these responses to 5-MeODMT. Responses to tryptamine and to apomorphine were enhanced ipsilateral to the lesion by 10- and 3-fold respectively. Haloperidol (0.5 mg/kg, i.v.) reversed apomorphine-induced inhibition of caudatal neuronal firing rate. The effects of tryptamine were not reversed by haloperidol. These data indicate a classical adaptive increase in [3H]tryptamine binding in caudate following 6-OHDA lesions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563337 TI - Eastern equine encephalitis in Quebec and Connecticut, 1972: introduction by infected mosquitoes on the wind? AB - In 1972 there were outbreaks of eastern equine encephalitis in the Eastern Townships, Quebec, Canada and in Connecticut, USA. Climatic data including Northern Hemisphere synoptic charts were examined. The findings indicate that the virus could have been brought to Lac Brome by infected mosquitoes carried on surface winds from Meriden, Connecticut, on the night of August 22-23, 1972. The distance of 400 km would have been covered in 14-16 h at a speed of 25-30 km h-1 and at a temperature of 15 degrees C and higher. The first case was recorded 13 days later on September 5, 1972. The outbreak at Meriden, Connecticut started on August 21, 1972. On August 7, 1972 southwesterly winds blew along the Atlantic coast at heights up to 1.5 km. Infected mosquitoes could have been carried on the wind from Cape May, New Jersey, Delaware-Maryland-Virginia peninsula, North Carolina or Georgia. Flights would have been at 17 degrees-20 degrees C and lasted 5-6, 9-10, 14-16 and 20-26 h depending on the origin. The arrival on August 8, 1972 coincided with a cold front moving from the northwest through Connecticut. Culiseta melanura is regarded as the mosquito species most likely to have been involved in the transmission of infection. PMID- 2563336 TI - Requirement for capsular antigen KX105 and fimbrial antigen CS1541 in the pathogenicity of porcine enterotoxigenic Escherichia coli O8:KX105 strains. AB - The requirement for capsular antigen KX105 and fimbrial antigen CS1541 in the pathogenicity of porcine enterotoxigenic Escherichia coli O8:KX105 strains lacking the colonization factor antigens K88, K99, 987P and F41 was investigated using two encapsulated strains and their acapsular variants, one of which produced the fimbrial antigen CS1541 in vitro. None of the strains adhered in vitro to enterocytes isolated from newborn colostrum-deprived piglets. All of the strains caused diarrhea in orally infected, hysterotomy-derived, colostrum deprived piglets although a great variability in the clinical response of the piglets was observed. Colonization of the small intestine of infected piglets by these strains was only moderate and no differences in the ability to colonize the small intestine was noted between the strains. All of the strains reacted in the indirect fluorescent antibody test with both CS1541 and 987P antisera when applied to organisms in the intestines of infected piglets. A control strain expressing the 987P fimbrial adhesin also reacted with the CS1541 antiserum applied to organisms in the intestines of an infected piglet. It was concluded that capsular antigen KX105 was not essential for intestinal colonization and production of diarrhea in hysterotomy-derived colostrum-deprived pigs, and that fimbrial antigen CS1541 does not promote in vitro adherence to enterocyte brush borders but could be important in bacterial colonization in vivo. PMID- 2563339 TI - Influence of beta-adrenoceptor blocking drugs on lipid-protein interaction in synaptosomal membranes. An ESR study. AB - The influence of the beta-adrenoceptor blocking drugs atenolol, doberol, propranolol and exaprolol on synaptosomal membranes was studied using ESR spectroscopy of stearic acid spin labeled at the 16th position. The drugs changed the ESR spectra of the label in the membranes, where in addition to changes of a fluid lipid component they increased the proportion of a motionally-restricted component. No motionally-restricted component was found in the samples prepared from brain total lipid liposomes treated with the drugs. The drug propensities at 20 mmol/l concentration to increase the proportion of the motionally-restricted component in the following order, control less than doberol approximately atenolol less than or equal to propranolol less than exaprolol did not correlate with their potency to influence the dynamics of the bulk lipid membrane phase. The motionally-restricted component induced by exaprolol increased with raising temperature and prolongation of time of the sample incubation. The results indicate that the beta-adrenoceptor blocking drugs influence lipid-protein interaction in the synaptosomal membranes, which could be important for elucidation of their mechanism of biological membrane activities. PMID- 2563338 TI - Percutaneous transluminal angioplasty for innominate artery stenosis and total occlusion of subclavian artery in Takayasu's-type arteritis. AB - Previous reports describe high success rates achieved by dilating subclavian artery stenoses. Attempts at angioplasty for total occlusions have been uniformly unsuccessful. No previous case successfully recanalizing total subclavian artery occlusion was found after an extensive literature search. Modified guidewire technique facilitated safe crossing of the occlusion. PMID- 2563340 TI - Does vecuronium accumulate in the renal transplant patient? AB - Twenty ASA physical status Class III patients undergoing cadaver renal transplantation were studied. After 90 per cent T1 recovery, as determined by train-of-four measurement, from 1.0 mg.kg-1 succinylcholine to facilitate tracheal intubation, nine patients received atracurium 0.25 mg.kg-1 (Group I) and 11 patients received vecuronium 0.05 mg.kg-1 (Group II) intravenously. The following measurements were made: time to maximum block onset (first dose Max), injection to start of recovery (start REC1), injection to 25 per cent T1 twitch recovery (REC 251), injection to 75 per cent T1 (REC 75(1], injection to 90 per cent T1 (REC 90(1] and time from 25-75 per cent recovery T1 (REC 25-75(1]. Maximum blockade (Max block 1) was also measured. At 90 per cent T1 recovery, if time permitted, an identical dose of the appropriate relaxant was administered. Time from second dose to onset of maximum block (second dose Max) and 90 per cent recovery after second dose (REC 90(2] were then measured. At the conclusion of surgery, neuromuscular blockade was reversed with neostigmine 2.5 mg and glycopyrrolate 0.5 mg. One way ANOVA was performed to determine significance between the groups and a p less than 0.05 was considered significant. A paired t test was also performed between REC 90(1) and REC 90(2) for atracurium and vecuronium respectively. A p less than 0.05 was again considered significant. Measurement of first dose Max, start REC1, REC25(1), REC 75(1), REC 90(1), REC 25 75(1) and Max block 1 revealed no difference between the patients receiving an initial dose of atracurium and those receiving vecuronium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563342 TI - Enhanced hypoxic pulmonary vasoconstriction in hypertension. AB - In this study, we tested the hypothesis that hypoxic pulmonary vasoconstriction may be enhanced in systemic hypertension. The hypothesis took origin from the following two considerations: alveolar hypoxia constricts the pulmonary vessels by enhancing the Ca2+ penetration across sarcolemma of the smooth muscle cells and systemic high blood pressure is associated with an elevation of tone and reactivity of the lung vessels, which seems to depend on an excessive cytosol free Ca2+ concentration due to alterations in sodium handling and in the Na+-Ca2+ exchange system. These considerations suggest the possibility that the disorders in the biochemistry of smooth muscle contraction in hypertension facilitate the rise of cytosol Ca2+ concentration during alveolar hypoxia, thus resulting in a potentiation of the vasoconstrictor properties of this stimulus. In 43 hypertensive and 17 normotensive men, pulmonary arteriolar resistance has been evaluated during air respiration and after 15 minutes of breathing 17%, 15%, and 12% oxygen in nitrogen. Curves relating changes in pulmonary arteriolar resistance to oxygen breathing contents had similar configuration in the two populations but in hypertension were steeper and significantly shifted to the left, reflecting a lower threshold and an enhanced reactivity. This pattern was not related to differences in severity of the hypoxic stimulus, plasma catecholamine concentration, or hypocapnia and respiratory alkalosis induced by hypoxia and probably was not mediated through alpha-receptor activation. Calcium channel blockade with nifedipine was able to almost abolish both the normotensive and the hypertensive pulmonary vasoconstriction reaction. These findings support the hypothesis that hypoxic pulmonary vasoconstriction may be enhanced in systemic hypertension. PMID- 2563341 TI - Monoamine oxidase inhibitors revisited. PMID- 2563343 TI - Continuous monitoring of mixed venous oxygen saturation as an indicator of pharmacologic intervention. AB - This prospective study evaluated the ability of continuous SvO2 measurements to predict the onset and duration of action of the oral vasodilator, fenoldopam. Eight patients with New York Heart Association functional class 3 CHF received 100 mg fenoldopam in the fasted state. Serial hemodynamic parameters and SvO2 measurements were obtained at baseline and up to eight hours postdose. Although wide interpatient variability was observed, the SvO2-time response curve produced a similar trend as the CI-time response curve. The SvO2 may be a useful drug monitoring parameter in patients with NYHA class 3 CHF. Seriously ill patients may not have a good CI/SvO2 correlation. This is most likely due to an unstable oxygen consumption rate at the tissue level. Therefore, we recommend establishing the relationship between CI and SvO2 prior to its use as a drug monitoring parameter. PMID- 2563344 TI - Experience with topical administration of 4-aminosalicylic acid in ulcerative colitis. AB - 4-Aminosalicylic acid was applied topically in a daily dose of 1.4 gm for two weeks in ten patients with ulcerative colitis. After favorable results, the therapeutic effects of 4-aminosalicylic acid and salazopyrin enemas were compared in a two-week cross-over open trial, in 20 patients suffering from recurrent ulcerative colitis involving the rectum and rectosigmoid. No significant difference was found in the changes of the endoscopic picture of the mucosa. The results did not show a significant difference between 4-aminosalicylic acid and salazopyrin enemas, either in the clinical activity or in the histologic picture. 4-Aminosalicylic acid seems to be a suitable drug for improving the clinical symptoms of ulcerative proctitis. PMID- 2563345 TI - Heterotopic gastric mucosa of the rectum--characterization of endocrine and mucin producing cells by immunocytochemistry and lectin histochemistry. Report of a case. AB - Heterotopic gastric mucosa in the rectum is particularly uncommon; only 23 cases have been reported to date. Moreover, no studies have been done on the neuroendocrine apparatus and glycoprotein production of the heterotopic mucosa. This study reports on a 13-year-old boy, admitted with rectal bleeding and persistent tenesmus. An ulcerative lesion was found on colonoscopy; biopsies revealed a fundic-type gastric tissue. Medical therapy (H2-blockers) promptly healed the rectal ulcer; surgical excision of the heterotopia was performed with complete and permanent relief of symptoms (3-year follow-up). Immunocytochemistry (PAP) revealed 5-Ht and somatostatin cells in the gastric-type mucosa, as in the normal human stomach. These cells also were present in the surrounding rectal epithelium where PYY-enteroglucagon cells were detected, which were absent in the heterotopic tissue. Mucin histochemistry showed PAS-positive cells also strongly stained by LA lectin in the heterotopic tissue, differentiating the rectal epithelium that remained unstained. Therefore, the morphofunctional status (endocrine cells and mucins) of the gastric heterotopia was almost identical to its orthotopic counterpart, confirming the hypothesis that endocrine cells and mucin-producing cells differentiate their metabolic products according to the anatomic and functional activity of the epithelium where they grow. PMID- 2563346 TI - Elimination of permeability mutants from selections for drug resistance in mammalian cells. AB - Chinese hamster ovary (CHO) cells exhibit increased sensitivity to a wide variety of microtubule inhibitory drugs when verapamil is present in the growth medium. The extent of this increased sensitivity is drug specific: some drugs such as taxol and vinblastine respond greatly to the presence of verapamil, whereas other drugs such as griseofulvin respond very poorly. For the majority of drugs examined, however, a 2- to 10-fold increase in drug sensitivity is observed in the presence of verapamil at 5 micrograms/ml. The effects of verapamil are even more dramatic when drug-resistant mutant cells with a presumed alteration in membrane permeability are examined. In the presence of appropriate levels of verapamil, these mutants demonstrate a level of drug sensitivity comparable to that of the wild-type parental cells. Drug-resistant cells from similar selections but with well-defined alterations in alpha- or beta-tubulin and no evidence of alterations in membrane permeability, however, continue to exhibit increased resistance to the selecting drug even in the presence of verapamil. These studies support the conclusion that verapamil affects the membrane permeability to or transport of a wide variety of hydrophobic drugs. In addition, we have used this information to devise selections that virtually eliminate the isolation of drug-resistant permeability mutants. This methodology should be generally applicable to genetic studies of drug action that are complicated by the isolation of large numbers of mutants with permeability alterations. PMID- 2563347 TI - Sulfasalazine metabolites and dapsone attenuate formyl-methionyl-leucyl phenylalanine-induced mucosal injury in rat ileum. AB - The effects of 5-aminosalicylic acid (5-ASA), 4-ASA, N-acetyl-5-ASA, and sulfapyridine on mucosal permeability were determined in an experimental model of acute ileitis. In addition, the antiinflammatory drug dapsone was tested. The distal 10 cm of rat ileum was perfused with formyl-methionyl-leucyl-phenylalanine (FMLP) (10(-5) M), a bacterial peptide that activates and attracts neutrophils. Changes in mucosal permeability were assessed using the blood-to-lumen clearance of 51Cr-ethylene-diamineacetate. Luminal FMLP increased 51Cr-labeled ethylenediamineacetate clearance twofold and fourfold in the first and second hour, respectively. Addition of 5-ASA (10 mM), 4-ASA (10 mM), or dapsone (4 mM) to the luminal perfusate after 60 min of FMLP perfusion greatly attenuated the increased mucosal permeability observed after 120 min of FMLP perfusion. Neither N-acetyl-5-ASA (10 mM) nor sulfapyridine (5 mM) had an effect on the FMLP-induced increase in mucosal permeability. We characterized the inhibitory effect of these drugs on the catalytic activity of myeloperoxidase and tested their ability to scavenge hypochlorous acid in vitro. 5-Aminosalicylic acid, 4-ASA, and dapsone demonstrated a powerful inhibitory effect on the catalytic activity of myeloperoxidase, whereas all drugs were equally effective in scavenging HOCl. In additional in vitro experiments we were unable to demonstrate an inhibitory effect of either of the drugs on the catalytic activity of neutrophilic elastase. Our results indicate that inhibition of neutrophilic myeloperoxidase may be an important mechanism by which 5-ASA, 4-ASA, and dapsone attenuate FMLP-induced mucosal injury. PMID- 2563349 TI - A genetic linkage map of five marker loci in and around the Duchenne muscular dystrophy locus. AB - Linkage analysis of five marker loci in and around the Duchenne muscular dystrophy (DMD) locus, DXS84, DXS206, DXS164, DXS270, and DXS28, was conducted with 499 families. Overall, the best multipoint distances were found to be DXS84 3.7 +/- 0.6 cM-DXS206-1.0 +/- 0.4 cM-DXS164-1.9 +/- 0.6 cM-DXS270-12.0 +/- 1.1 cM DXS28. A comparison of this linkage map with the established physical map suggests the presence of hot spots for recombination in the DMD locus. PMID- 2563348 TI - A genetic linkage map of the long arm of human chromosome 22. AB - We have used a recombinant phage library enriched for chromosome 22 sequences to isolate and characterize eight anonymous DNA probes detecting restriction fragment length polymorphisms on this autosome. These were used in conjunction with eight previously reported loci, including the genes BCR, IGLV, and PDGFB, four anonymous DNA markers, and the P1 blood group antigen, to construct a linkage map for chromosome 22. The linkage group is surprisingly large, spanning 97 cM on the long arm of the chromosome. There are no large gaps in the map; the largest intermarker interval is 14 cM. Unlike several other chromosomes, little overall difference was observed for sex-specific recombination rates on chromosome 22. The availability of a genetic map will facilitate investigation of chromosome 22 rearrangements in such disorders as cat eye syndrome and DiGeorge syndrome, deletions in acoustic neuroma and meningioma, and translocations in Ewing sarcoma. This defined set of linked markers will also permit testing chromosome 22 for the presence of particular disease genes by family studies and should immediately support more precise mapping and identification of flanking markers for NF2, the defective gene causing bilateral acoustic neurofibromatosis. PMID- 2563350 TI - Confirmation of linkage of Friedreich ataxia to chromosome 9 and identification of a new closely linked marker. AB - A linkage analysis with chromosome 9 markers was performed in 33 families with Friedreich ataxia (FA). Linkage with D9S15, previously established by S. Chamberlain et al. (1988, Nature London 334:248-249) was confirmed in our sample (z(theta) = 6.82 at theta = 0.02) while INFB (interferon-beta gene) shows looser linkage. An additional marker, D9S5, was also shown to be closely linked to FA (z(theta) = 5.77 at theta = 0.00). PMID- 2563351 TI - Chromosomal mapping of the mouse IL-4 and human IL-5 genes. AB - We mapped the mouse interleukin (IL)-4 gene on chromosome 11 by restriction fragment length polymorphism using recombinant inbred mouse strains. The human IL 5 gene was mapped on chromosome 5q 23.3-31.1 by in situ hybridization. Because the granulocyte macrophage colony-stimulating factor (GM-CSF) and IL-3 genes were previously mapped on mouse chromosome 11 (within a 230-kb region) and human chromosome 5, the IL-4 and IL-5 genes are likely to cluster on the same chromosomes with the GM-CSF and IL-3 genes in both species. PMID- 2563352 TI - X-chromosome inactivation in the Wiskott-Aldrich syndrome: a marker for detection of the carrier state and identification of cell lineages expressing the gene defect. AB - Recently developed techniques for the direct analysis of DNA have made possible the determination of patterns of cellular X-chromosome inactivation. These techniques provide a potential method for carrier detection for several X-linked human disorders in which obligate carriers show nonrandom X inactivation. By using restriction fragment length polymorphic (RFLP) gene-specific probes in conjunction with methylation-sensitive enzymes, we have characterized the patterns of X-chromosome inactivation in cell subsets from females belonging to 10 kindreds segregating for the X-linked immune deficiency disorder Wiskott Aldrich syndrome (WAS). We show that selective inactivation of the X chromosome distinguishes obligate WAS carriers from noncarrier females and constitutes a valuable marker of the WAS carrier state. Selective inactivation phenomena were observed in the monocytes and T and B lymphocytes of obligate carriers, implying that the WAS gene defect is expressed in each of these cellular lineages. In conjunction with the use of linked DNA markers, RFLP-methylation analysis should render carrier detection feasible for the majority of females from WAS families. The results of such analyses also provide an initial step toward identifying the cellular level and molecular basis for WAS. PMID- 2563353 TI - Mapping of the human complement factor I gene to 4q25. AB - A detailed genetic and physical map of human complement factor I (IF) using somatic cell hybrids, in situ hybridization, and genetic linkage is reported. The gene has been localized to band 4q25. The order GC-INP10-ADH3-EGF-IF-IL2-MNS is proposed for genes on 4q on the basis of genetic and physical mapping techniques. A BclI polymorphism found with the IF probe demonstrated a maternal origin for a de novo deletion of chromosome 4 that was used in physically mapping the gene. The genetic and physical distances around band 4q24 suggest that 1 cM is approximately 1.2 million bp of DNA. This work provides a useful addition to the map of 4q. PMID- 2563354 TI - Restriction fragment length polymorphism in canine narcolepsy. PMID- 2563355 TI - Molecular genotyping of four chicken B-complex haplotypes with B-L beta, B-F, and B-G probes. PMID- 2563356 TI - Linkage of the mouse Hsp84 heat shock protein structural gene to the H-2 complex. PMID- 2563357 TI - Complexity, polymorphism, and connectivity of mouse Vk gene families. AB - To define the polymorphism and extent of the mouse immunoglobulin kappa (Igk) gene complex, we have analyzed restriction-enzyme digested genomic DNA from 33 inbred strains of mice with labeled DNA probes corresponding to 16 Vk protein groups (1 of them previously undescribed) and the Jk/Ck region (V, variable; J, joining; C, constant). These probes detected between 1 and 25 distinct restriction enzyme fragments (REF) that appeared in up to eight polymorphic patterns, thus defining eight mouse Igk haplotypes. The investigated portion of the Vk repertoire was estimated to encompass between 60 and 120 discernable Vk gene-containing REFs. In contrast to mouse VH gene families, several Vk gene families defined by these probes appeared to overlap. This observation has implications for Vk gene analyses by nucleic acid hybridization and raises the possibility that the Vk gene complex is a continuum of related sequences. PMID- 2563358 TI - The mouse Igh-1a and Igh-1b H chain constant regions are derived from two distinct isotypic genes. AB - Genetic and structural analyses of the mouse genes encoding constant region of immunoglobulin subclass (Igh-C) have shown that recombination is rare within this cluster which is inherited as a set designated the Igh haplotype. Recent molecular analyses have demonstrated that either DNA exchanges or gene duplications have probably occurred during the evolution of this set of genes. In order to assess the generality of the duplication processes, the presence and expression of two allelic forms of the Igh-1 (gamma 2a) gene (Igh-1a and Igh-1b) were examined in a large panel of wild mice belonging to Mus musculus domesticus and Mus musculus musculus species. Our data indicate that certain M. m. domesticus animals and most animals in the M. m. musculus group coexpress the two allelic forms of Igh-1. Moreover, genetic studies show that these two immunoglobulin types are encoded by tandemly arranged genes. We propose that wild mice, from which laboratory mice are derived, carry three isotypic gamma 2 genes (Igh-1a, Igh-1b, Igh-3), and these have given rise to the two isotypes seen in laboratory strains by a deletion/insertion mechanism. PMID- 2563359 TI - Biting rhythm of the vectors of Malayan filariasis, Mansonia annulifera, M. uniformis & M. indiana in Shertallai (Kerala state), India. AB - Biting activity of Mansonioides mosquitoes through all night collections for a period of two years in Shertallai region, in south India was studied. M. annulifera bites uniformly throughout the night, with two peaks of activity, one after midnight (24.00-01.00 h) and another before dawn (04.00-05.00 h). M. uniformis was found to be predominantly dusk biting, with its peak of activity between 1800 to 1900 h. M. indiana showed two peaks of biting activity one after dusk (2000-2100 h) and another before dawn (0300-0400 h). All biting, with its peak of activity between 1800 to 1900 h. M. indiana showed two peaks of biting activity one after dusk (2000-2100 h) and another before dawn (0300-0400 h). All these species following a particular pattern of activity cycles discerned the biting rhythm of Mansonioides mosquitoes to be species-specific. PMID- 2563360 TI - Definition of IDDM-associated HLA DQ and DX RFLPs by segregation analysis of multiplex sibships. AB - Genetic variation of the DQ alpha and beta and of the DX alpha genes, detectable as RFLP in genomic DNA digests, has been suggested to improve the identification of individuals at high risk for insulin-dependent diabetes mellitus (IDDM). DNA from all members of 32 IDDM multiplex families was digested with six restriction endonucleases and the resulting fragments analyzed in Southern blots for hybridization with labeled cDNA probes for those genes. A computerized segregation analysis procedure was then used to assign fragments to haplotypes. Associations among fragments and between fragments and haplotypes characterized serologically and biochemically for their class II genes and IDDM-carrier status were calculated. The results indicate that the alleles of the DX alpha polymorphism maintain linkage disequilibrium with those of the DQ beta genes responsible for the well-known DQ beta 3.2-IDDM association, so that IDDM-carrier haplotypes carry disproportionally often both DQ beta 3.2 and DX alpha-TaqI 2.2kb. Thus, these RFLPs identify a DR-DQ-DX haplotype in high linkage disequilibrium, rather than the locus or loci that account for their high relative risk. However, four DR4-DQ beta 3.2 haplotypes that lack DX alpha-TaqI 2.2kb were encountered, two of which are "affected." These haplotypes suggest that the identification of the "disease locus" can be facilitated by the study of unusual haplotypes in which distinct IDDM-associated alleles occur separated from their neighbors of the standard genetic configurations. PMID- 2563361 TI - Stimulant medication and the retarded. PMID- 2563362 TI - Mechanism of pneumococcal cell wall degradation in vitro and in vivo. AB - We compared the products of autolytic amidase-catalyzed wall degradation in vivo (in penicillin-induced lysis) and in vitro. Pneumococci labeled in their cell wall stem peptides by radioactive lysine were treated with penicillin, and the nature of wall degradation products released to the medium during lysis of the bacteria was determined. At early times of lysis (20% loss of wall label), virtually all the radioactive peptides released (greater than 94%) were of high molecular size and were still attached to glycan and teichoic acid. At times of more extensive bacterial lysis (56%), progressively larger and larger fractions of the released peptides became free, i.e., detached from glycan and teichoic acid. Analysis of the nondegraded residual wall material by high-resolution high pressure liquid chromatography revealed that this in vivo-triggered autolysis did not involve selective hydrolysis of some of the chemically distinct stem peptides. Parallel in vitro experiments yielded completely different results. Purified pneumococcal cell walls labeled with radioactive lysine were treated in vitro with low concentrations of pure amidase, and the nature of wall degradation products released during limited hydrolysis and after more extensive degradation was determined. In sharp contrast to the in vivo experiments, the main products of in vitro hydrolysis were free peptides. After a short treatment with amidase (resulting in a 20% loss of label), the material released was enriched for the monomeric stem peptides. At all times of hydrolysis (including the time of extensive degradation), only a relatively small fraction of the released wall peptides was covalently attached to glycan and teichoic acid components (17% as compared with 40% in the intact cell wall). We propose that the in vivo-triggered amidase activity first attacks the amide bonds in some strategically located (or unprotected) stem peptides that hold large segments of cell wall material together. The observations indicate that the in vivo activity of the pneumococcal autolysin is under topographic constraints. PMID- 2563363 TI - Genetic relationship of two highly studied Synechococcus strains designated Anacystis nidulans. AB - The cyanobacteria Synechococcus sp. strain PCC 7942 and Synechococcus sp. strain PCC 6301 are very closely related and both have been designated by the binomial Anacystis nidulans. The only established difference between the two strains is the superior transformation properties of strain PCC 7942. Significant homology between the rRNA genes of these strains was demonstrated by the ability of an rRNA operon from strain PCC 6301, interrupted by a spectinomycin and streptomycin resistance marker, to transform strain PCC 7942 by recombining with and replacing an endogenous rRNA operon. Restriction fragment length polymorphism data indicated that the chromosomes of the two strains were conserved around the three psbA loci, the two rRNA operons, and the psbDI locus. However, multiple polymorphisms were detected downstream of the psbDII locus, identifying a DNA rearrangement such as an inversion, insertion, or deletion within the chromosome. Analysis of genome structure by pulsed-field gel electrophoresis of large NotI restriction fragments showed only two bands that were visibly shifted between the chromosomes of the two strains. These data support their very close genetic relationship and the feasibility of studying genes derived from strain PCC 6301 in the highly transformable PCC 7942 strain. PMID- 2563364 TI - Mechanism of L-glutamate transport in membrane vesicles from Bacillus stearothermophilus. AB - In the presence of electrochemical energy, several branched-chain neutral and acidic amino acids were found to accumulate in membrane vesicles of Bacillus stearothermophilus. The membrane vesicles contained a stereo-specific transport system for the acidic amino acids L-glutamate and L-aspartate, which could not translocate their respective amines, L-glutamine and L-asparagine. The transport system was thermostable (Ti = 70 degrees C) and showed highest activities at elevated temperatures (60 to 65 degrees C). The membrane potential or pH gradient could act as the driving force for L-glutamate uptake, which indicated that the transport process of L-glutamate is electrogenic and that protons are involved in the translocation process. The electrogenic character implies that the anionic L glutamate is cotransported with at least two monovalent cations. To determine the mechanistic stoichiometry of L-glutamate transport and the nature of the cotranslocated cations, the relationship between the components of the proton motive force and the chemical gradient of L-glutamate was investigated at different external pH values in the absence and presence of ionophores. In the presence of either a membrane potential or a pH gradient, the chemical gradient of L-glutamate was equivalent to that specific gradient at different pH values. These results cannot be explained by cotransport of L-glutamate with two protons, assuming thermodynamic equilibrium between the driving force for uptake and the chemical gradient of the substrate. To determine the character of the cotranslocated cations, L-glutamate uptake was monitored with artificial gradients. It was established that either the membrane potential, pH gradient, or chemical gradient of sodium ions could act as the driving force for L-glutamate uptake, which indicated that L-glutamate most likely is cotranslocated in symport with one proton and on sodium ion. PMID- 2563365 TI - Purification and properties of glutamine synthetase from the non-N2-fixing cyanobacterium Phormidium laminosum. AB - Soluble glutamine synthetase activity (L-glutamate:ammonia ligase, ADP forming, EC 6.3.1.2) was purified to electrophoretic homogeneity from the filamentous non N2-fixing cyanobacterium Phormidium laminosum (OH-1-p.Cl1) by using conventional purification procedures in the absence of stabilizing ligands. The pure enzyme showed a specific activity of 152 mumol of gamma-glutamylhydroxamate formed.min-1 (transferase activity), which corresponded to 4.4 mumol of Pi released.min-1 (biosynthetic activity). The relative molecular mass of the native enzyme was 602 kilodaltons and was composed of 12 identically sized subunits of 52 kilodaltons. Biosynthetic activity required the presence of Mg2+ as an essential activator, although Co2+ and Zn2+ were partially effective. The kinetics of activation by Mg2+, Co2+, and Zn2+ were sigmoidal, and concentrations required for half-maximal activity were 18 mM (h = 2.2), 6.3 mM (h = 5.6), and 6.3 mM (h = 2.45), respectively. However, transferase activity required Mn2+ (Ka = 3.5 microM), Cu2+, Co2+, or Mg2+ being less effective. The substrate affinities calculated for L-Glu, ammonium, ATP, L-Gln, and hydroxylamine were 15, 0.4, 1.9 (h = 0.75), 14, and 4.1 mM, respectively. Optimal pH and temperature were 7.2 and 55 degrees C for biosynthetic activity and 7.5 and 45 degrees C for transferase activity. The biosynthetic reaction mechanism proceeded according to an ordered three-reactant system, the binding order being ammonium, L-Glu, and ATP. The presence of Mn2+ or Mg2+ drastically affected the thermostability of transferase and biosynthetic activities. Heat inactivation of biosynthetic activity in the presence of Mn2+ obeyed first-order kinetics, with an Ea of 76.8 kcal (ca. 321 kJ) mol-1. Gly, L Asp, L-Ala, L-Ser and, with lower efficiency, L-Lys and L-Met, L-Lys, and L-Glu inhibited only transferase activity. No cumulative inhibition was observed when mixtures of amino acids were used. Biosynthetic activity was inhibited by AMP (Ki= 7 mM), ADP (Ki= 2.3 mM), p-hydroxymercuribenzoate (Ki= 25 microM), and L methionine-D, L-sulfoximine (Ki= 2 microM). The enzyme was not activated in vitro by chemically reduced Anabaena thioredoxin. This is the first report of glutamine synthetase activity purified from a filamentous non-N2-fixing cyanobacterium. PMID- 2563366 TI - Structure and function of conjugative pili: inducible synthesis of functional F pili by Escherichia coli K-12 containing a lac-tra operon fusion. AB - In vivo and in vitro recombination methods were used to construct the recombinant plasmid pTG801, in which the F-plasmid DNA transfer (tra) genes required for the formation of functional F pili were placed under the lac transcriptional control sequences of pUC19. The 20 kilobases of cloned F DNA includes genes traA through the 5'-terminal part of traG; the plasmid lacks the positive regulatory gene traJ and all of the known tra genes required for the DNA transfer stage of conjugation. pTG801 transformants were sensitive to the donor-specific bacteriophages Q beta and f1, as measured by the formation of infectious centers. They were relatively insensitive to bacteriophage R17, as expected from the absence of traD. In the presence of a lacIq allele, sensitivity of pTG801 transformants to f1 and Q beta depended on the concentration of inducer (isopropyl-beta-D-thiogalactopyranoside [IPTG]). Viewed by electron microscopy, pTG801 transformants elaborated 7- to 10-nm-diameter filaments that could be laterally decorated with RNA bacteriophage particles, consistent with the formation of F pili. In stationary-phase cultures, these filaments formed massive aggregates and could be seen to adhere lengthwise to the cell surface; few pili accumulated in the medium as single filaments. PMID- 2563367 TI - Transformation-deficient mutants of piliated Neisseria gonorrhoeae. AB - Seven transformation-deficient mutants of piliated, competent Neisseria gonorrhoeae were isolated by screening them for their inability to be transformed by chromosomal DNA after chemical mutagenesis. Three distinct classes of mutants were obtained, each of which was piliated, as determined by electron microscopy. One class exhibited abnormal colony morphology and was unable to take up DNA into a DNase-resistant state. A second class was morphologically normal and took up DNA into a DNase-resistant state normally, but was deficient in both chromosomal and plasmid transformation; mutations in these mutants may affect entry of DNA into the cell proper. A third class was similar to the second but was fully competent for plasmid transformation, suggesting that there was a defect in a late stage of chromosomal transformation. PMID- 2563368 TI - In vitro protein synthesis by the moderate halophile Vibrio costicola: site of action of Cl- ions. AB - In vitro protein synthesis in Vibrio costicola [poly(U)-directed incorporation of phenylalanine] was studied. The extent of protein synthesis was limited by the number of ribosomes present. Density gradient centrifugation experiments suggested that, after runoff of ribosomes from the artificial messenger, the 50S subunit was unable to attach to the 30S-messenger complex. As shown previously (M. Kamekura and D. J. Kushner, J. Bacteriol. 160:385-390, 1984), Cl- ions inhibited protein synthesis; indeed, the highest rate of synthesis took place in the lowest attainable Cl- concentration (37 mM). The inhibitory effects were partly reversed by glutamate and betaine, both of which are concentrated within cells of V. costicola. The strongest reversal was seen when both glutamate and betaine were present. Cl- ions can prevent binding of ribosomes to poly(U) and displace ribosomes already bound to this artificial messenger. The effects of Cl- ions on binding were also reversed by glutamate and betaine. Cl- ions did not affect accuracy of translation; they were shown previously (Kamekura and Kushner, J. Bacteriol. 160:385-390, 1984) not to affect phenylalanyl-tRNA synthetase. It was also found that washing ribosomes with inhibitory NaCl concentrations did not interfere with their ability to carry out protein synthesis later in optimal (low) salt concentrations. On the contrary, these ribosomes were more active than before they were washed. We conclude that the main site of action of Cl- in the system studied is on the binding of ribosomes to the mRNA. PMID- 2563369 TI - Decrease of anion selectivity caused by mutation of Thr501 and Gly502 to Glu in the hydrophobic domain of the colicin E1 channel. AB - Structure-function relations of the colicin E1 ion channel were studied through the effects of mutations in the 35-residue hydrophobic region of the channel polypeptide and neighboring residues in the channel domain. Mutation of neutral residues threonine 501 and glycine 502 to a more polar or charged glutamic acid generated a protein whose channel conductance properties in each case had a decreased selectivity for anions. There was no significant effect on ion selectivity caused by mutations that changed residue charge outside the hydrophobic domain at the neighboring aspartic acid 509 or at glycine 439. The Thr501----Glu and Gly502----Glu mutants possessed lower cytotoxic and in vitro activity. An altered thermolysin cleavage pattern and a greater binding to membrane vesicles at pH greater than 4.5 of the Gly502----Glu mutant indicated greater exposure of its COOH-terminal hydrophobic domain in solution. It is concluded that the hydrophobic nature of threonine 501 and glycine 502 is important in the structure of the channel lumen and the soluble colicin. Altering proline 462, a residue conserved in five sequenced channel-forming colicins, had no significant effect on channel properties. These conclusions are discussed in the context of sequence-structure-function concepts for channel proteins. PMID- 2563370 TI - Genomic organization and polymorphisms of the human C3d/Epstein-Barr virus receptor. AB - The human C3d/Epstein-Barr virus receptor (CR2/CD21) is a 145-kDa protein primarily expressed on mature B lymphocytes. CR2 is a member of the regulators of complement activation (RCA) gene family found on band q32 of chromosome 1. The RCA proteins are characterized by the presence of 60-70 amino acid short consensus repeats (SCR). A full length CR2 cDNA was cloned and used to identify overlapping cosmid genomic clones. Analysis of CR2 exon-intron junctions revealed the presence of three types of exons in the short consensus repeat region of CR2. First, four exons each of which encodes two SCR are present. Five exons encode a single SCR. Six exons encode SCRs which are split in identical positions. The order of these types of exons is in a repeated array of four SCRs, indicating that the contemporary CR2 gene likely evolved from a more primitive gene containing four SCRs. The CR2 full length cDNA clone was used to find restriction fragment length polymorphisms (RFLPs). Restriction enzyme TaqI generated 2.55- and 2.10-kilobase (kb) polymorphic bands. This RFLP was mapped near the exon containing the first two SCRs. HaeIII digestion generated polymorphic bands of 1.45, 1.55, and 1.75 kb. The HaeIII 1.45-kb RFLP band maps near the exon containing the 15th SCR. The TaqI and HaeIII RFLPs will provide tools for the genetic analysis of CR2. The organization of the CR2 gene provides insights into the evolution of human CR2 and the RCA gene family. PMID- 2563371 TI - Isolation of tyrosine-melanocyte-stimulating hormone release-inhibiting factor 1 from bovine brain tissue. AB - Although Tyr-melanocyte-stimulating hormone release-inhibiting factor 1 (MIF-1) (Tyr-Pro-Leu-Gly-NH2) can exert a number of biological actions in the brain and elsewhere, it has never been isolated from any tissue. Accordingly, we attempted to purify it from acetic acid extracts of bovine brain tissue by gel filtration chromatography and several different high performance liquid chromatographic systems. Peptide content was followed by a specific and sensitive radioimmunoassay with an antibody that was generated against synthetic Tyr-MIF-1. In each of the five applied high performance liquid chromatographic systems, the immunoreactive fractions coincided exactly with the elution time of synthetic Tyr MIF-1 in the control runnings. The structure of the isolated peptide was identified by microsequence analysis as the tetrapeptide Tyr-Pro-Leu-Gly-NH2 and shown to be biologically active. PMID- 2563372 TI - Characterization of a large form of DNA polymerase delta from HeLa cells that is insensitive to proliferating cell nuclear antigen. AB - A large form of DNA polymerase delta from HeLa cells was recently purified in this laboratory as a factor required for conservative DNA synthesis in a reconstituted system utilizing UV-irradiated permeabilized human diploid fibroblasts (Nishida, C., Reinhard, P., and Linn, S. (1988) J. Biol. Chem. 263, 501-510). We have now purified this form of the enzyme utilizing its polymerase activity and further characterized it. The enzyme activity sediments at 11.1 S in low salt and 6.8 S in high salt. In both cases, activity cosediments with the major visible peptide displayed by sodium dodecyl sulfate-polyacrylamide gels which has an Mr of 215,000. This value is consistent with the molecular mass calculated from the sedimentation coefficient and gel filtration behavior in high salt. In low salt the apparent molecular mass was approximately double. The enzyme prefers poly(dA).oligo(dT) as template/primer in low salt, with which it has a processivity of several thousand nucleotides in 1 mM MgCl2. At isotonic KCl or potassium phosphate concentrations, the preferred template/primer is activated DNA. Proliferating cell nuclear antigen, also characterized as a DNA polymerase delta auxiliary protein, does not increase the activity of this preparation of the enzyme. An antibody to the proliferating cell nuclear antigen has no inhibitory effect, nor is it able to recognize any peptides in immunoblots of purified enzyme fractions. Under polymerizing conditions, the enzyme removes mismatched, but not matched nucleotides from the 3' terminus of oligo(dT) annealed to poly(dA) suggesting a proofreading function. The properties of this form of DNA polymerase delta distinguish it from other preparations reported in the literature. PMID- 2563373 TI - Misplacement of the amino-terminal positive charge in the prepro-alpha-factor signal peptide disrupts membrane translocation in vivo. AB - We have identified a series of mutations in the signal peptide of yeast prepro alpha-factor which specifically attenuate translocation across the endoplasmic reticulum membrane in vivo. In prepro-alpha-factor-somatostatin hybrids, transposition of the amino-terminal tripeptide from wild-type NH2-Met-Arg-Phe to NH2-Met-Phe-Lys or NH2-Met-Phe-Arg causes a 45-75% reduction in the efficiency of membrane translocation. This is evidenced by the intracellular accumulation of unglycosylated, signal-containing precursors which are membrane-associated and are exposed to the cytosol. Surprisingly, abolition of the single positive charge by replacing arginine with phenylalanine has little effect on translocation into the endoplasmic reticulum. We conclude that the presence of an amino-terminal positive charge is not necessary for efficient targeting or translocation; however, misplacement by one position markedly disrupts translocation without affecting targeting. These mutations thus define an early stage of membrane interaction that is sensitive to local charge effects. Furthermore, our data suggest that post-translational translocation, signal cleavage, and core glycosylation of these polypeptides may occur to a significant extent in vivo. PMID- 2563374 TI - Disruption of the 290-342 salt bridge is not responsible for the secretory defect of the PiZ alpha 1-antitrypsin variant. AB - Crystallographic studies have previously suggested that Lys290 forms a salt bridge with Glu342 in the serine protease inhibitor alpha 1-antitrypsin. Disruption of the formation of this structural feature by a Glu to Lys substitution at residue 342 in the PiZ variant has been implicated in causing the defective secretion of this mutant protein from hepatocytes (10-15% of normal). To test the validity of this hypothesis, mutant human alpha 1-antitrypsin cDNA constructs coding for specific amino acid substitutions at residues 290 and 342 were generated and the corresponding mutant proteins were expressed in mouse hepatoma cells. When the potential to form the salt bridge was reestablished by a Lys290 to Glu290 substitution in the PiZ variant, its secretion was increased to only 38% of normal. Furthermore, disruption of this structural feature by a Lys290 to Glu290 substitution in the normal inhibitor failed to reduce the secretion of alpha 1-antitrypsin to the extent observed for the PiZ variant (73% of normal). Finally, substitution of the neutral amino acid Gln at residue 342 only reduced the secretion of alpha 1-antitrypsin to 55% of normal. Of all mutant proteins tested, those bearing Lys at position 342 were secreted at the lowest levels. These findings demonstrate that although disruption of the 290-342 salt bridge does affect the secretion of alpha 1-antitrypsin, it is the substitution of Lys at residue 342 that causes the dramatic secretory defect of the PiZ variant. PMID- 2563375 TI - Dermorphin gene sequence peptide with high affinity and selectivity for delta opioid receptors. AB - Skin of the frog Phyllomedusa sauvagei contains a cDNA sequence that codes for the selective mu-receptor peptide dermorphin and a new heptapeptide we have designated as dermorphin gene-associated peptide (DGAP). Investigation of the opioid receptor binding characteristics of synthetic DGAP and [D-Met2]DGAP revealed that the latter peptide had high affinity and selectivity for delta-type opioid receptors in rat brain synaptosomes. The IC50 values for DGAP on mu- and delta-receptors were only 28 microM and 670 nM, respectively, while that for [D Met2]DGAP was 0.80 nM for delta-receptors and greater than 1 microM for mu receptors yielding a very high delta selectivity ratio (SR) of 1345. In comparison, the SR values for [D-Ala2,D-Leu5]enkephalin, [D Ser2,Leu5,Thr6]enkephalin, and [D-Pen2,5]enkephalin, ligands which are considered to be specific for delta-receptors, were 20, 42, and 301, respectively. Dermorphin, which contains a D-Ala2 residue and is a selective mu-receptor ligand (Lazarus, L.H., Guglietta, A., Wilson, W.E., Irons, B.J., and de Castiglione, R. (1989) J. Biol. Chem. 264, 354-362), exhibits a SR of 0.0055 similar to that for the conventional mu-agonist [D-Ala2,NMePhe4,Gly-ol]enkephalin (0.0040). This finding that frog skin cDNA contains the information to code for dermorphin and DGAP, or the presumed [D-Met2]DGAP molecule, which are among the most selective high affinity opioid ligands described for mu- and delta-receptors, may permit new insight into the design of future opioid receptor agonists and antagonists. PMID- 2563376 TI - CpG mutations in the reactive site of human C1 inhibitor. AB - C1 inhibitor plays an important role in the regulation of vascular permeability through its ability to inactivate enzymes which release polypeptide kinins. Dysfunctional C1 inhibitor molecules are present in the plasma of affected members of the Da and Ri hereditary angioneurotic edema kindreds. We constructed genomic libraries from Da and Ri patient DNAs which had been cleaved with BclI to generate a fragment containing 21 kilobases of the C1 inhibitor locus. C1 inhibitor gene-containing recombinants originating from mutant Da and Ri alleles were differentiated from those derived from normal alleles by linkage analysis using the intragenic HgiAI restriction fragment length polymorphism. Nucleotide sequencing of the complete protein-coding regions of the mutant alleles identified two different mutations in a CpG dinucleotide corresponding to the first two bases of arginine codon 444. These single base mutations changed the identity of the functionally critical P1 reactive site residue from arginine to cysteine (Da) or histidine (Ri). The additional cysteine residue in C1 inhibitor Da suggests how it is covalently bound to albumin in plasma. The presence of CpG dinucleotides in the codons specifying the P1 arginines of C1 inhibitor and antithrombin III explains the high incidence of histidine and cysteine substitutions observed among dysfunctional mutants of these serine protease inhibitors. PMID- 2563377 TI - Study of the 5-oxoprolinase reaction by 13C NMR. AB - 5-Oxoprolinase catalyzes the ATP-dependent decyclization of 5-oxo-L-proline to L glutamate. Previous studies provided evidence for the intermediate formation of a phosphorylated form of 5-oxoproline (Seddon, A. P., and Meister, A. (1986) J. Biol. Chem. 261, 11538-11541) and of a tetrahedral intermediate (Li, L., Seddon, A. P., and Meister, A. (1987) J. Biol. Chem. 262, 11020-11025). A new approach to the study of the reaction mechanism using the 18O isotope effect on the 13C NMR signals for 5-oxoproline and glutamate is reported here. The 13C chemical shifts induced by 18O substitution for the carbonyl group of 5-oxoproline and the gamma carboxyl group of glutamate are about 0.03 ppm with respect to the corresponding 16O-compounds. Using 5-[18O]oxo[5-13C]proline (97 and 79.5 atom % excess, 13C and 18O, respectively), the disappearance of the 18O-labeled and unlabeled 5 oxoproline and formation of the corresponding glutamate species were followed in the reactions catalyzed by purified preparations of 5-oxoprolinase isolated from Pseudomonas putida and from rat kidney. This procedure permits simultaneous determinations of the rates of 18O exchange and of the overall decyclization reaction. The ratios of 18O exchange rates to the overall reaction rates for the bacterial and kidney enzyme catalyzed-reactions were 0.28 and 0.14, respectively. The findings support the view that the coupling of ATP hydrolysis to 5-oxoproline decyclization involves formation of a phosphorylated tetrahedal intermediate. Although the exchange phenomena are consistent with the mechanistic interpretations, they seem not to be required for catalysis. PMID- 2563378 TI - Newly synthesized calsequestrin, destined for the sarcoplasmic reticulum, is contained in early/intermediate Golgi-derived clathrin-coated vesicles. AB - We have examined the possible role of clathrin-coated vesicles (CVs) in the genesis of the sarcoplasmic reticulum (SR) in developing chick skeletal myotubes. Calsequestrin (CSQ) a luminal Ca2+ binding protein of the terminal SR cisternae, is contained within the vesicle lumen of skeletal muscle CVs in substantial amounts, approximately four molecules/CV. Employing 3-day cultures of chick skeletal myotubes we demonstrate that after a 30-min labeling with [35S]methionine and cysteine, radioactivity in CSQ remains high in the CVs 45 min later and then declines, while labeled CSQ in the SR continues to rise. No CSQ appears to be secreted. All of the CSQ in both the CVs and SR is sensitive to the activity of endoglycosidase H, and a significant fraction also binds to wheat germ agglutinin. Based on these results, we discuss the hypothesis that a selective CV-mediated pathway exists in developing skeletal muscle cells for the transport of CSQ from the early/intermediate Golgi apparatus to the SR. PMID- 2563379 TI - Intracellular transport, sorting, and turnover of acetylcholinesterase. Evidence for an endoglycosidase H-sensitive form in Golgi apparatus, sarcoplasmic reticulum, and clathrin-coated vesicles and its rapid degradation by a non lysosomal mechanism. AB - Tissue-cultured muscle cells synthesize several oligomeric forms of acetylcholinesterase (AChE) destined for the cell surface or secretion. Previous studies on the biogenesis of AChE polypeptide chains have shown that only a small fraction become assembled into catalytically active oligomers which transit the Golgi apparatus and acquire endoglycosidase H (endo H) resistance. Most of the AChE polypeptides remain endo H-sensitive and are rapidly degraded intracellularly. We now show that all newly synthesized AChE polypeptides are transported from the rough endoplasmic reticulum to the Golgi apparatus where they acquire N-acetylglucosamine. However, approximately 80% of these AChE polypeptides remain endo H-sensitive and are degraded intracellularly with a half life of about 1.5 h by a mechanism which is insensitive to lysosomotropic agents. These endo H-sensitive AChE molecules can be chased into clathrin-coated vesicles and/or the sarcoplasmic reticulum prior to degradation. Pulse-chase studies of isotopically labeled or catalytically active AChE molecules suggest that there are at least two discreet populations of clathrin-coated vesicles which leave the Golgi, one whose origin is cis/medial and one whose origin is trans. These studies indicate the existence of a post-rough endoplasmic reticulum, non lysosomal degradative pathway for intra-luminal proteins and suggest that post translational events at the levels of protein sorting and degradation may play a role in regulating the abundance of exportable proteins. PMID- 2563380 TI - Conversion of amino acid residues in proteins and amino acid homopolymers to carbonyl derivatives by metal-catalyzed oxidation reactions. AB - A number of metal-catalyzed oxidation (MCO) systems mediate the oxidative inactivation of enzymes. This oxidation is accompanied by conversion of the side chains of some amino acid residues to carbonyl derivatives (for review, see Stadtman, E. R. (1986) Trends Biochem. Sci. 11, 11-12). To identify the amino acid residues which are sensitive to MCO oxidation, several enzymes/proteins and amino acid homopolymers were exposed to various MCO systems. The carbonyl groups which were formed were converted to their corresponding 3H-labeled hydroxy derivatives. After acid hydrolysis, the labeled free amino acids were separated by ion exchange chromatography. Each protein or polymer gave rise to several different labeled amino acids. The elution profiles of the labeled amino acids obtained from preparations of Escherichia coli glutamine synthetase which had been oxidized by MCO systems comprised of either Fe(II)/O2 or ascorbate/Fe(II)/O2 both in the presence and absence of EDTA were qualitatively the same. From a comparison of the elution profiles of labeled amino acids from various proteins with those obtained from homopolymers, it is evident that the side chains of histidine, arginine, lysine, and proline are particularly sensitive to oxidation by the MCO systems. This conclusion is supported also by direct amino acid analysis of acid hydrolysates which shows that the oxidation of glutamine synthetase, enolase, and phosphoglycerate kinase is associated with the loss of at least 1 histidine residue per subunit. From the results of studies with homopolymers, it is apparent that glutamic semialdehyde is a major product of both proline and arginine residues. In addition, hydroxyproline and unlabeled glutamic acid were identified among the hydrolysis products of oxidized poly-L proline, and unlabeled aspartic acid was identified as a product of poly-L histidine oxidation. PMID- 2563381 TI - Chromosomal protein HMG-14. Identification, characterization, and chromosome localization of a functional gene from the large human multigene family. AB - The human HMG-14 multigene family is one of the largest retropseudogene families known. To identify and isolate a functional human HMG-14 gene, genomic clones, selected with the cDNA, were screened with a set of 6 oligonucleotides. A single genomic clone was isolated suggesting that the human genome contains few, and perhaps only one, functional genes. An 8882-base pair (bp) genomic clone containing the complete, 6804-bp-long human gene together with 850 bp 5' to the start of transcription and 1228 bp 3' to the end of transcription was sequenced. The gene is comprised of 6 exons ranging in size from 30 to 839 bp, two of which code for the entire DNA binding site of the protein, and has several features typical of "housekeeping" genes. Using human-rodent somatic cell hybrids, the HMG 14 gene was localized to human chromosome 21. A restriction fragment length polymorphism, useful for further analysis and mapping, has been detected. The present article, which describes the first isolation and characterization of a gene coding for chromosomal protein HMG-14, indicates that genes coding for HMG 14 and HMG-17 may share several distinctive characteristics. Comparison with the human and chicken HMG-17 genes reveals that all contain 6 exons, that all have exons of similar size, that all have 5' regions highly enriched in GC residues and that all have features typical of housekeeping genes. PMID- 2563382 TI - Primary structure of rat liver dipeptidyl peptidase IV deduced from its cDNA and identification of the NH2-terminal signal sequence as the membrane-anchoring domain. AB - Two forms of dipeptidyl peptidase IV (DPP) were purified from rat liver plasma membranes: a membrane form (mDPP) extracted with Triton X-100 and a soluble form (sDPP) prepared by treatment with papain. Apparent molecular masses of mDPP and sDPP were 109 and 105 kDa, respectively, when determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The NH2-terminal sequences of the two forms were found to be completely different from each other. For further information on the molecular structure, we constructed a lambda gt11 liver cDNA library and isolated two cDNA clones for DPP, lambda cDP37 and lambda cD5. The 3.5-kilobase cDNA insert of lambda cDP37 contains an open reading frame that encodes a 767-residue polypeptide with a calculated size of 88,107 Da, which is in reasonable agreement with that of DPP (87 kDa) immunoprecipitated from cell free translation products. Eight potential N-linked glycosylation sites were found in the molecule, accounting for the difference in mass between the precursor and mature forms. Of particular interest is that the deduced NH2 terminal sequence with a characteristic signal peptide is completely identical to that determined for mDPP. In addition, the NH2-terminal sequence of sDPP is identified in the predicted sequence starting at the 35th position from the NH2 terminus. These results indicate that the signal peptide of DPP is not cleaved off during biosynthesis but functions as the membrane-anchoring domain even in the mature form. It is also found that the primary structure thus predicted has striking homology to that of gp 110, a bile canaliculus domain-specific membrane glycoprotein (Hong, W., and Doyle, D. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 7962-7966). PMID- 2563383 TI - Prevalence of Escherichia coli strains with localized, diffuse, and aggregative adherence to HeLa cells in infants with diarrhea and matched controls. AB - To determine the possible role of Escherichia coli strains with three different patterns of adherence to HeLa cells in causing diarrhea in infants in Sao Paulo, Brazil, we studied stool specimens from 100 infants up to 1 year of age with acute diarrheal illnesses and 100 age-matched control infants without recent diarrhea. E. coli with localized adherence to HeLa cells was much more common in patients (23%) than in controls (2%) (P less than 0.0001) and was detected more frequently than rotavirus (19%) was in patients, even though the study was conducted during the coldest months of the year. Most (80%) of the E. coli colonies with localized adherence were of traditional enteropathogenic E. coli serotypes. Little difference was found between patients and controls in the rate of isolation of E. coli with diffuse adherence (31 and 32%, respectively) or aggregative adherence (10 and 8%, respectively). A genetic probe used to detect a plasmid-mediated adhesin which confers expression of localized adherence proved to be 100% sensitive and 99.9% specific in detecting E. coli with localized adherence to HeLa cells. Although E. coli strains with localized adherence have now been shown to be enteric pathogens in several parts of the world, the role of strains showing diffuse adherence and aggregative adherence is still uncertain. PMID- 2563384 TI - Genetic diversity within Streptococcus mutans evident from chromosomal DNA restriction fragment polymorphisms. AB - Attempts to study the acquisition, transmission, and other aspects of the natural history of Streptococcus mutans infections in humans have been hampered by limitations and inconsistencies in methods by which phenotypic characteristics of individual isolates are examined. Because most mutans streptococci associated with human dental caries fall within the biotype I (serotypes c and f) grouping, designated S. mutans, these typing methods are of little value in distinguishing individual isolates. Here we show that strains of S. mutans obtained from over 30 individuals demonstrate unique "fingerprints" of chromosomal DNA digested with restriction endonuclease HaeIII. To demonstrate that this polymorphism in restriction fragments can be used to study the acquisition and transmission of this organism, we examined isolates of S. mutans from three mother-infant pairs obtained at the time the infant first became colonized by this organism. Results indicate that strains of S. mutans found in infants exhibit restriction fragment profiles identical to those of their mothers, strongly supporting the notion that mothers transmit this organism to their infants. Also, we show that strains of S. mutans with the same restriction fragment profile were stably maintained over a 3 year interval in the one mother-infant pair studied. Moreover, we found that mothers and their infants harbored only a few individual strains, suggesting that transmission of this organism is probably confined within discrete family cohorts. Collectively, these findings demonstrate the potential utility of genomic fingerprinting in studying the natural history of S. mutans infections in humans. PMID- 2563385 TI - Examination of colonies and stool blots for detection of enteropathogens by DNA hybridization with eight DNA probes. AB - We compared three methods for detecting enteropathogens in 416 children with diarrhea: (i) examination of 10 lactose-fermenting and all non-lactose-fermenting Escherichia coli (colony blots); (ii) examination of 300 colonies (replicate blots); and (iii) determination of the total bacterial growth of stools (stool blots). All specimens were spotted onto Whatman 541 filters and hybridized with specific radiolabeled DNA probes. Enterotoxigenic E. coli was detected in 38 patients by examining colony blots, in 52 patients by examining replicate blots, and in 45 patients by examining stool blots. Enteropathogenic E. coli adhesin factor was detected in 12 patients by colony blots, in 25 patients by replicate blots, and in 16 patients by stool blots. E. coli that hybridized with the enterohemorrhagic E. coli probe was detected in 2 patients by colony blots, in 11 patients by replicate blots, and in 0 patients by stool blots. Shiga-like toxin producing E. coli was detected in 0 patients by colony blots, in 12 patients by replicate blots, and in 0 patients by stool blots. Shigella spp. were identified by standard bacteriological methods in 82 patients, and enteroinvasive E. coli was identified by colony blots in 11 patients (total, 93), by replicate blots in 56 patients, and by stool blots in 35 patients. Of 82 culture-confirmed Shigella infections, 45 were identified by examining replicate blots with the 17-kilobase pair probe and 36 were identified by examination with the Ipa probe (P less than 0.05). Examining replicate blots with specific probes identified more enterotoxigenic E.coli (P < 0.005), enteropathogenic E.coli adhesion factor producing E.coli (P < 0.001), and Shiga-like toxin-producing E.coli (P < 0.005) infections than examining colony blots. More Shigella and enteroinvasive E.coli infections were identified by standard bacteriological methods and examining colony blots with a specific probe than by examining replicate and stool blots (P < 0.001). PMID- 2563386 TI - DNA probe for identification of Streptococcus pneumoniae. AB - A total of 287 clinical isolates of Streptococcus pneumoniae (pneumococcus) were tested for their ability to undergo autolysis when treated with sodium deoxycholate. The test was positive for all but one isolate, strain DOC-1. This autolysis required the activity of an enzyme which is unique and characteristic of S. pneumoniae: a choline-dependent N-acetylmuramoyl-L-alanine amidase, the gene product of the lytA gene. We used lytA as a DNA probe to test the distribution of the autolysin gene among clinical isolates of S. pneumoniae. In dot blot hybridization experiments our probe reacted with the DNA of 60 of 60 strains tested, including the autolysis-deficient clinical isolate DOC-1. No hybridization occurred when strains of Streptococcus sanguis, Streptococcus mutans, Streptococcus pyogenes, Streptococcus (Enterococcus) faecalis, Streptococcus (Enterococcus) faecium, Streptococcus agalactiae, and Streptococcus bovis were tested. The lytA gene appears to be an ideal candidate for use as a DNA probe for the identification of S. pneumoniae. PMID- 2563388 TI - The use of a DNA probe for epidemiological studies of candidiasis in immunocompromised hosts. AB - Reproducible typing procedures to differentiate isolates of Candida albicans are limited. C. albicans isolates were obtained from immunocompromised patients by using DNA restriction enzyme fragment analysis and hybridization with both a radiolabeled mitochondrial DNA probe and a nonradioactive (biotinylated) DNA probe. There were 110 pathogenic and nonpathogenic C. albicans isolates from 63 immunocompromised patients. EcoRI restriction fragment analysis with the biotinylated probe revealed different "fingerprint" patterns for 60 of 63 patients. Analysis of 57 isolates from 20 patients showed no intrapatient variation regardless of the isolation site. DNA probe "fingerprint" patterns were analyzed for eight patients on serially recovered (range, 2-18 mo) C. albicans isolates. The unique patient profiles persisted over time. The application of this biotinylated C. albicans DNA probe provides a more sensitive means than simple gel restriction fragment analysis to define the epidemiology of C. albicans infection. The use of this biotin-labeled nonradioactive probe has potential application in clinical evaluations of outbreaks of nosocomial candidiasis. PMID- 2563387 TI - The murine Il-6 gene maps to the proximal region of chromosome 5. AB - Murine Il-6 cDNAs were isolated by cross-hybridization with a human IL-6 cDNA from an IL-1 activated bone marrow stromal cell line (W20). Mouse-hamster somatic cell hybrids were utilized to localize murine Il-6 to chromosome 5. Genetic mapping with respect to En-2, AlbH, and Gus in backcross progeny from an interspecific mating between C57BL/6J and Mus spretus positioned Il-6 3 cM distal to En-2. The syntenic relationships of Il-6 and En-2 in mouse and man, as well as the potential role of IL-6 in tumorigenesis, are discussed. PMID- 2563390 TI - Priming of polymorphonuclear neutrophilic leukocyte oxidative activity by type 1 pili from Escherichia coli. AB - Preincubation of human polymorphonuclear leukocytes with free whole type 1 pili derived from Escherichia coli or with subunits of type 1 pili significantly augmented subsequent stimulation of cellular oxygen uptake by formyl-methionyl leucyl-phenylalanine, concanavalin A, opsonized zymosan, or latex particles coated with whole type 1 pili. No such priming of leukocyte activation occurred if phorbol myristate acetate was used as a stimulant or if bovine serum albumin was substituted for the type 1 pili preparations. When free in solution, neither pili preparation directly stimulated cellular oxidative metabolism. Type 1 pili mediated priming was not inhibited by polymyxin B, by mannose congeners, or by boiling the pili preparations but was prevented by trypsinization of the pili. This priming property indicates a further means by which type 1 pili may modulate the inflammatory response of the host and provides another tool that can be used to dissect the relationship between the structure and function of type 1 pili and to characterize leukocyte receptors for these pili. PMID- 2563389 TI - DNA polymorphisms among Escherichia coli isolated from bacteriuric women. AB - Restriction fragment length polymorphism (RFLP) patterns, plasmid profiles, and antimicrobial susceptibility patterns of paired sequential Escherichia coli isolates from antibiotic-treated and untreated elderly women were analyzed. Isolates from 26 of 27 subjects who were treated successfully with antibiotics but became reinfected differed by RFLP analysis, whereas 10 of 12 subjects who failed treatment and 11 of 14 untreated subjects had paired isolates with identical RFLP banding patterns. Only 40 of the 53 pairs of isolates could be analyzed by plasmid profiles; 36 of these 40 were concordant with RFLP analysis. Antimicrobial susceptibility patterns showed poor concordance with RFLP analysis. This study demonstrates the utility of RFLP analysis and indicates that isolation of E. coli from elderly women after sterilization of the urinary tract usually results from introduction of a new strain; elderly women who fail antibiotic therapy or receive no therapy may remain persistently infected with the same E. coli strain. PMID- 2563391 TI - Prevalence of the putative colonization factors CFA/III and PCFO159:H4 in enterotoxigenic Escherichia coli. PMID- 2563392 TI - Weiss lecture. Radiation chemistry: basic, strategic or tactical science? AB - The work of Weiss in the 1930s, particularly with Haber, has only recently been recognized to have implications in biology and medicine. Similarly, research in radiation chemistry and the application of the pulse radiolysis technique, for example, have implications far beyond traditional radiation chemistry. Some examples of such research are discussed against a background of categorization into 'basic', 'strategic' or 'tactical' science, rather than 'pure' and 'applied'. Examples discussed include redox properties of free radicals, which are now on a firm quantitative basis, and the identification and characterization of nitro radicals as intermediates in drug metabolism. Radical reactions often take place in multicomponent systems, and the techniques of radiation chemistry can be used to probe, for example, events occurring at interfaces in micelles. Industrial processes involving radiation are attracting investment, particularly in Japan. Radiation chemistry deserves further support to exploit its full potential, but much research using radiation chemical techniques often appears, probably more appropriately, under different labels. PMID- 2563393 TI - The radiation-induced inactivation of external yeast invertase in dilute aqueous solution. AB - The inactivation of external yeast invertase by irradiation in dilute aqueous solution has been investigated. The contributions of the individual radical species from water radiolysis to inactivation and amino acid degradation were estimated from the results of experiments in which solutions were saturated with nitrogen, nitrous oxide or oxygen, and on addition of hydroxyl radical scavengers. Under conditions where inactivation by hydroxyl radicals predominates, the rate of inactivation increased with increasing dose, indicating that in the initial stages of the radiolysis the mannose-rich oligosaccharide chains of the glycoprotein protect the polypeptide chain from radical attack. Amino acid analysis of the irradiated external invertase showed that there was significant destruction of tyrosine, phenylalanine, methionine and histidine residues. Destruction of methionine and histidine residues may be responsible for the free radical-induced inactivation of this enzyme. PMID- 2563394 TI - Cellular glutathione content and K values. II. AB - After exposure to radiation in the presence of different oxygen concentrations, the OER values were determined for human fibroblasts cell strains with differing intrinsic GSH content, and cells depleted of GSH to different extent by treatment with BSO. The yield of DNA single-strand breaks was used as the measure of radiosensitivity. The K values were found to be related specifically to the GSH concentration in the cell nuclei. A good agreement was found between the experimental observations and the prediction of a variant of the competition model in which a second type of damage, not influenced by oxygen, was included. PMID- 2563395 TI - Breakage of human interphase chromosomes by alpha particles and X-rays. AB - The technique of premature chromosome condensation (PCC) was used to compare the early formation of chromosome breaks in non-cycling HF19 human diploid fibroblasts when irradiated with slow alpha particles (3.2 MeV, 128 keV micron-1) or 250 kVP X-rays. For both radiations the production of PCC breaks increased approximately linearly with dose. The production coefficient for alpha particles was 12.5 +/- 0.6 per cent per Gy and for X-rays it was 5.8 +/- 0.2 per cell per Gy. Hence, the relative biological effectiveness (RBE) of the alpha particles was 2.16 +/- 0.13. This is smaller than reported values of the RBE for the production of chromosome-type exchange aberrations by slow alpha particles. This implies that there is a difference, spatial or qualitative, in the initial breaks produced by the densely ionizing alpha particle tracks and the more sparsely ionizing electron tracks from the X-rays. PMID- 2563396 TI - Enhanced sensitivity to camptothecin in ataxia-telangiectasia cells and its relationship with the expression of DNA topoisomerase I. AB - The antitumour drug camptothecin (CPT) can trap covalently bound topoisomerase I DNA intermediates as complexes which conceal single-strand scissions. In an attempt to evaluate the cytotoxic potential of these lesions in human cells we have measured: (1) cell cycle delay and cell killing by CPT in primary and transformed fibroblasts, and in lymphoblastoid lines derived from normal, X-ray sensitive ataxia-telangiectasia (A-T) and xeroderma pigmentosum (XP) donors; (2) the properties of sublines obtained by high-dose selection in CPT: (3) levels of drug-induced DNA strand scission in intact cells; (4) the cellular availability of extractable topoisomerase I. The drug induced a marked cell cycle block in G2 phase, the magnitude of the block being closely related to cell kill. XP group A cells showed normal sensitivity to CPT, whereas A-T derived cells were consistently hypersensitive (3-5 fold) in a manner which could not be related to a primary deficiency in topoisomerase I activity, abnormal capacity for complex formation or anomalies in the intracellular generation of DNA strand breaks. A CPT-resistant A-T subline had reduced topoisomerase I activity but retained the characteristic of hypersensitivity to X-radiation. The subline lost resistance upon in vitro passage with evidence that resistance was initially an unstable feature of a subpopulation of cells. The findings have implications for the role of topoisomerase I in the in vitro phenotype of A-T cells, and the contribution made by topoisomerase I-dependent damage to the cytotoxic action of CPT. PMID- 2563397 TI - Changes in X-ray sensitivity of mouse eggs from fertilization to the early pronuclear stage, and their repair capacity. AB - To study the changes in radiosensitivity of male and female genomes from fertilization to the pronuclear stage, the frequency of induced chromosome aberrations was examined at the first-cleavage metaphase in eggs fertilized with X-irradiated sperm, in eggs X-irradiated at the mature oocyte stage immediately before fertilization, and in fertilized eggs exposed to X-rays at various stages before DNA synthesis (1-5 h after insemination). Gametic treatment, fertilization and embryo culture were carried out in vitro. Most of the induced chromosome aberrations were chromosome-type aberrations, the frequency of chromosome fragments being the highest, followed by chromosome exchanges in both sperm and oocytes. The induction of chromosome-type aberrations was much higher in oocytes than in sperm. Chromosome-type aberrations were the main type also in fertilized eggs irradiated at the pre-DNA-synthetic stage. The radiosensitivity increased gradually with pronuclear formation (1-4h after insemination), but little difference in radiosensitivity was observed between eggs irradiated at 4 h and 5 h, corresponding to the stage when pronuclear formation was complete. The drastic change with pronuclear formation was found not only in radiosensitivity but also in the frequency of chromosome aberrations. The frequency of chromosome-type exchanges decreased drastically 2 h after insemination, and exchanges were barely observed at 5 h. The difference in radiosensitivity between male and female genomes also markedly changed with pronuclear formation, the chromosome aberration induction in the female genome being much higher than that in the male genome before accomplishment of pronuclear formation. The analysis of potentiation effects of 3-aminobenzamide and caffeine on the yield of X-ray induced chromosome aberrations demonstrated that the increase of radiosensitivity and the decrease of chromosome-type exchange induction with pronuclear formation, may be closely correlated with alterations in chromatin configuration in the pronuclei and in repair capacity of fertilized eggs at the pre-DNA-synthetic stage. No evidence based on repair efficiency was found for the marked difference in radiosensitivity between male and female genomes during pronuclear formation. PMID- 2563398 TI - Radiation-induced increase in the release of amino acids by isolated, perfused skeletal muscle. AB - Local exposure of the hindquarter of the rat to 15 Gy of gamma-radiation resulted, 4-6 h after irradiation, in an increased release of amino acids by the isolated, perfused hindquarter preparation, 70 per cent of which is skeletal muscle. This increase in release involves not only alanine and glutamine which are synthesized to a large extent de novo in muscle, but also those amino acids which are not metabolized by muscle and, therefore, released in proportion to their occurrence in muscle proteins. Because metabolic parameters and content of energy-rich phosphate compounds in muscle remain unchanged, it is unlikely that general cellular damage is the underlying cause of the radiation-induced increase in amino acid release. The findings strongly favour the hypothesis that the increased availability of amino acids results from enhanced protein breakdown in skeletal muscle which has its onset shortly after irradiation. This radiation induced disturbance in protein metabolism might be one of the pathogenetic factors in the aetiology of radiation myopathy. PMID- 2563399 TI - Time course of loss of residual radiation damage in murine skin assessed by retreatment. AB - The amount of radiation damage remaining in mouse foot skin has been assessed by retreatment from 10 days to 6 months after a range of first doses. The acute skin reaction was used as the endpoint. Mice hind feet were first irradiated with a range of single doses (15-37.5 Gy) covering zero to near full effect. Feet were retreated with a full range of single doses together with groups of non previously treated age-matched control mice. No age-related changes in radiation sensitivity were observed. Dose-response curves were constructed for all retreatment times for each priming dose, and isoeffect doses were calculated for both peak and average skin reactions. If 2-6 months were allowed to elapse before retreatment, the skin could be reirradiated as if it were previously untreated. However, if only 1 month was allowed to pass before retreatment, damage was 'remembered' after all first doses. The amount of damage 'remembered' in terms of dose was 11 Gy after a first dose of 37.5 Gy, and was less after the lower first doses. PMID- 2563400 TI - Pulmonary immune response of dogs after exposure to 239PuO2. AB - This study evaluated the cell-mediated (CMI) and humoral immune responses in four Beagle dogs five to six years after single inhalation exposures to different monodisperse 239PuO2 aerosols (0.72-1.4 microns activity median aerodynamic diameter). These exposures resulted in initial lung burdens ranging from 19 to 35 kBq. Four nonexposed dogs were used as age-matched controls. Anesthetized dogs were immunized by instillation of sheep red blood cells (SRBC) into selected lung lobes. Cells and fluids were obtained serially from blood samples and by bronchoalveolar lavage of the saline- and SRBC-treated lung lobes at 5-20 days after immunization. The CMI response evaluated by the leukocyte procoagulant activity test was similar in the saline- and SRBC-treated lobes of both groups of dogs. The humoral immune response was measured by the enzyme-linked immunosorbent assay. No differences were shown between the amount of antibody measured in the sera or lung lavages from control or Pu-exposed dogs. Histopathology of the tracheobronchial lymph nodes from the Pu-exposed dogs showed them to be fibrotic with no lymphoid cells, suggesting that these tissues could not respond to the antigen deposited in the lungs. However, both mediastinal and sternal lymph nodes did contain lymphoid tissue, and were likely to be the lymphoid tissues that produced the immunity to the antigen deposited in the lungs of the exposed dogs. Although both exposed and control dogs produced immune responses to the antigen instilled into their lungs, differences were observed in the number of neutrophils in lung lavages from the control and exposed animals. There was a dramatic influx of neutrophils into both the saline- and SRBC-treated lung lobes of the Pu-exposed dogs that was not seen in the age-matched controls. This suggests that the inhaled 239PuO2 produced chronically-active inflammation in the lung which may contribute to recruitment of lymphocytes to the lung following intrapulmonary deposition of antigen. In conclusion, the immune responses induced by lung immunization of dogs that had inhaled 239PuO2 were not suppressed by large doses of chronic alpha irradiation of the lungs and tracheobronchial lymph nodes, indicating that local pulmonary immune responses are preserved despite severe radiation-induced alteration of these tissues. PMID- 2563401 TI - Incorporation of [3H]acetate into the membrane lipids of a murine tumour during the development of thermotolerance. AB - The incorporation of [3H]acetate into the membrane lipids of a C3H mammary adenocarcinoma, grown s.c. in the hind paw of CBA mice, was followed to estimate the effects on the de novo synthesis of membrane lipids after hyperthermic treatments. Thermotolerance developed in response to a heat treatment at 43 degrees C for 20 min, as verified through growth rate studies of tumours exposed to fractionated heat treatments. Our results show that, during the development of thermotolerance, the relative rates of incorporation of [3H]acetate into the major lipid classes of the tumour cell membranes change significantly. The de novo synthesis of phospholipids decreased while that of cholesteryl esters plus triglycerides increased. The incorporation of [3H]acetate into cholesterol remained constant. Consequently, the ratio [3H]cholesterol/[3H]lecithin increased significantly during the development of thermotolerance. When the incorporation of [3H]acetate was followed 72-96 h after the heat treatment, i.e. at the interval at which heat resistance was observed to approach that of control tumours, the incorporation into cholesterol was significantly reduced while incorporation into phospholipids increased to control levels. Thus, the ratio [3H]cholesterol/[3H]lecithin was significantly lower, when compared to that of control tumours. The functional relationship between the heat-induced changes in the de novo synthesis of membrane lipids and the development of thermotolerance is discussed with regard to a mechanism based on homeoviscous adaptation of the membranes. PMID- 2563402 TI - Advanced drug therapy for juvenile rheumatoid arthritis. AB - Childhood rheumatic diseases are frequently chronic, painful, and potentially debilitating. They may adversely affect growth and development, compromise future quality of life, and contribute added stress to the patient and family. Awareness of these consequences provides a stimulus to develop more effective therapeutic regimens. There is optimism that new therapeutic strategies will result in the more widespread and earlier use of drugs, including those discussed, that may substantially impede or arrest the underlying disease. PMID- 2563404 TI - Evaluation of xylazine hydrochloride as the sole immobilizing agent in moose and caribou--and its subsequent reversal with idazoxan. AB - Xylazine hydrochloride was used as the sole immobilizing agent in moose and caribou. The animals were free-ranging and immobilization was accomplished from a helicopter using powered darts. Following a period of immobilization during which radiotelemetry collars were fitted, the animals were revived using idazoxan (RX 781094) or its methoxy analogue RX 821002. Xylazine was administered at dose rates of approximately 3.0 mg/kg and 5.0 mg/kg to the moose and caribou, respectively. Moose received 430 +/- 27 mg of xylazine and a mean dose of 10 mg idazoxan (RX 781094). Caribou received 485 +/- 30 mg xylazine and a mean dose of 4 mg idazoxan (RX 821002). This technique gave adequate immobilization with rapid recovery of consciousness in both species. PMID- 2563405 TI - AMA-MSS Interim Meeting. PMID- 2563403 TI - Expression of lacto series type 2 antigens in human renal cell carcinoma and its clinical significance. AB - We performed immunohistochemical examination of serial sections of human fetal and adult renal tissue as well as human renal carcinoma tissue, using monoclonal antibodies T5A7, 1B2, FH2, FH4, and FH6. These monoclonal antibodies were directed to lacto series type 2 antigens with sugar-chain structures: lactosylceramide, lactoneotetraosylceramide (paragloboside), Lex (a chemically well-defined fucosyl carbohydrate antigen), difucosyl Lex, and sialosyl-difucosyl Lex, respectively. The staining pattern in fetal renal tissue changed significantly according to the stage of organogenesis. In addition, expression of the antigens, especially paragloboside and sialosyl-difucosyl Lex, was closely related to the prognosis of the patient. These results suggest that the expression of a series of oncofetal antigens in development or differentiation of organs is reflected in the reversion to an immature pattern of antigenic expression in tumor tissue. The pattern of antigen expression in renal tumors offers a good criterion for ascertaining the degree of tumor differentiation and malignancy and is valuable for determining prognosis. PMID- 2563406 TI - [Multiple endocrine neoplasia]. PMID- 2563407 TI - Acceleration of a murine T-cell leukemia associated with loss of interleukin-3 producing activity. AB - L-8313 is a murine T-cell leukemia cell line the cells of which constitutively produce interleukin-3 (IL-3). S2-8313 is a unique subline of L-8313 which has lost IL-3 producing activity. Although the growth of S2-8313 cells in the bone marrow is comparable to that of L-8313 cells, the survival time of C3H mice grafted with S2-8313 cells is significantly shorter than that of C3H mice grafted with L-8313 cells. The accelerated death observed in C3H mice bearing S2-8313 cells is attributable to early development of granulocytopenia and thrombocytopenia, which resulted from the loss of the IL-3 producing activity. PMID- 2563408 TI - Poorly expressed CD2 antigen on the leukemic cells of adult T-cell leukemia and chronic lymphocytic leukemia of T-cell lineage. AB - Adult T-cell leukemia (ATL) and T-cell chronic lymphocytic leukemia (T-CLL) are hematologic neoplasms in differentiated stages of peripheral mature T cells. This is suggested by the presence of CD2 (E rosette receptor) and mature and/or pan-T cell membrane surface antigens on their leukemic cells. We recently encountered one patient with ATL and another with T-CLL; their leukemic cells poorly expressed CD2 antigens, but clinical presentation, morphology of the leukemic cells and other marker studies were characteristic of either ATL or CLL. The clinical significance of the poor expression of CD2 remains to be further studied. The two patients reported here died of severe complications 4 and 9 weeks after diagnosis. The poor expression of CD2 in the peripheral T cells in these neoplasms is likely to indicate an aggressive nature of the disease. PMID- 2563409 TI - Effect of parenteral glutamine peptide supplements on muscle glutamine loss and nitrogen balance after major surgery. AB - Twelve patients admitted for elective resection of carcinoma of colon or rectum were allocated at random to experimental and control groups (six in each) and received a total parenteral nutrition regimen providing 230 mg N/kg and 166 KJ/kg daily over the first 5 postoperative days. In the experimental group the parenteral fluid was supplemented with a synthetic glutamine-containing dipeptide, L-alanyl-L-glutamine (54 mg peptide-N/kg per day) and the control group received corresponding amounts of alanine-N and glycine-N. On each postoperative day nitrogen balance was better in the experimental group; mean daily nitrogen balance with alanyl-glutamine was -1.5 (SE 0.4) g N/day and with the control solution -3.6 (0.2) g N/day. The cumulative nitrogen balances on the fifth postoperative day were -7.1 (2.2) and -18.1 (1.7) g N, respectively. With the peptide-containing solution intramuscular glutamine concentration remained close to the preoperative value whereas with the control solution it decreased from 19.7 (SE 0.9) to 12.0 (0.6) mmol/l intracellular water. PMID- 2563410 TI - Tyramine conjugation test for prediction of treatment response in depressed patients. AB - In a group of 30 patients with major unipolar depressive disorder urinary excretion of tyramine sulphate was measured after an oral dose of tyramine. There was a significant correlation between impaired tyramine conjugation and response to tricyclic antidepressant medication. PMID- 2563411 TI - Duplications of mitochondrial DNA in mitochondrial myopathy. AB - Restriction enzyme analysis was done on total cellular DNA extracted from whole blood in two patients with mitochondrial myopathy and multisystem involvement and their families. The two patients had an abnormal mitochondrial genome with a large (about 8 kb) duplication present in several tissues. Normal mitochondrial DNA (mtDNA) was also present, but within each maternal lineage the abnormal mitochondrial genome was confined to clinically affected individuals. This observation, together with the failure of extensive population surveys to identify such abnormalities of mtDNA, suggests that these mutations cause the disease. PMID- 2563412 TI - Practical use of duplex Doppler analysis of the renal vasculature in critically ill patients. AB - Duplex doppler examination of blood flow in renal interlobar arteries was analysed in twelve critically ill patients before and during low-dose dopamine infusion (2 micrograms/kg/min). Renal vasodilatation and increased blood flow were observed with dopamine, confirming results with indirect or invasive techniques. The doppler ultrasound technique is entirely non-invasive, is simple and quick to carry out, provides instant results, and will allow tailoring of inotropic support in critically ill patients to provide optimum renal blood flow. PMID- 2563413 TI - Localisation of endocrine-related tumours with radioiodinated analogue of somatostatin. AB - Various endocrine-related tumours contain large numbers of high-affinity somatostatin receptors. 123I-labelled tyr-3-octreotide (tyr-3-SMS 201-995, a synthetic derivative of somatostatin) was used to localise such tumours in vivo with a gamma-camera. Positive scans were obtained for two meningiomas, two gastrinomas, and one carcinoid; negative scans were obtained for one insulinoma (in which unlabelled octreotide had no effect on insulin levels), one phaeochromocytoma, one adrenal carcinoma (octreotide had no effect on cortisol levels), and three medullary thyroid carcinomas (octreotide had no effect on calcitonin levels). Thus radioiodinated tyr-3-octreotide can label somatostatin receptors in endocrine-related tumours in vivo and can therefore be used for tumour localisation. PMID- 2563414 TI - Curtains up on the NHS review. PMID- 2563415 TI - Trends in lymphoma diagnosis. PMID- 2563416 TI - Mitochondrial DNA and genetic disease. PMID- 2563417 TI - Monitoring the anaesthetist. PMID- 2563418 TI - Lactating adenomas of the breast. PMID- 2563419 TI - Morbidity of very low birthweight infants at corrected age of two years in a geographically defined population. Report from Project on Preterm and Small for gestational age infants in The Netherlands. AB - In a nationwide prospective survey on very preterm and very-low-birthweight infants in the Netherlands, a neurodevelopmental assessment was made at the corrected age of two years in a virtually complete population. The study achieved a 97.4% follow-up rate. A major handicap was found in 59 children and a minor handicap in 111 children (4.4% and 8.3% of liveborn infants, respectively). Unlike mortality, handicap was apparently unrelated to gestational age or birthweight. PMID- 2563420 TI - Nephrotic syndrome: from toddlers to twenties. AB - 63 patients with steroid-sensitive, biopsy-proven minimal-change nephrotic syndrome were followed for between 10 and 21 years. 2 died. All the survivors had normal renal function and blood pressure, and only 2 had a single attack. Frequent relapse was more common with young age of onset and in boys. The frequency of relapse fell rapidly over the first 4 years after diagnosis and then plateaued. Relapses continued into adult life. No definite endpoint to the disease could be defined although there was a linear relation between length of remission and risk of subsequent relapse. PMID- 2563421 TI - Current status of allergen immunotherapy. Shortened version of a World Health Organisation/International Union of Immunological Societies Working Group Report. PMID- 2563422 TI - Treatment of neurocysticercosis: is praziquantel the new hope? PMID- 2563423 TI - Clinical freedom: two lessons for the UK from US experience with privatisation of health care. PMID- 2563424 TI - Aluminium and Alzheimer's disease. PMID- 2563425 TI - Neutrophil cytoplasmic autoantibodies and Wegener's granulomatosis. PMID- 2563426 TI - Brother-to-sister transmission of measles after measles, mumps, and rubella immunisation. PMID- 2563427 TI - Measles, mumps, and rubella vaccine coverage. PMID- 2563428 TI - Treatment of disseminated peritoneal hydatid disease with praziquantel. PMID- 2563429 TI - Carbon dioxide breathing and mountain sickness. PMID- 2563430 TI - Mental problems and frequent use of analgesics. PMID- 2563431 TI - Mosaicism and sporadic haemophilia: implications for carrier determination. PMID- 2563433 TI - Retinopathy of prematurity. PMID- 2563432 TI - Point mutation in c-HA-ras oncogene in normal and abnormal cells. PMID- 2563434 TI - Uncoated versus coated 5-aminosalicylic acid in Crohn's disease. PMID- 2563435 TI - Asymptomatic myocardial ischaemia: biobehavioural mechanism. PMID- 2563436 TI - Genetics of schizophrenia. PMID- 2563438 TI - Computers and general practice. PMID- 2563437 TI - Nottingham study of neurotic disorder. PMID- 2563439 TI - Criticism of IPPNW. PMID- 2563440 TI - No-fault compensation. PMID- 2563441 TI - Birth defect registries. PMID- 2563442 TI - Lack of serological association between herpesvirus and atherosclerosis. PMID- 2563443 TI - Parvovirus and idiopathic thrombocytopenic purpura. PMID- 2563444 TI - Epstein-Barr virus in brain and Rasmussen's encephalitis. PMID- 2563445 TI - Food irradiation. PMID- 2563446 TI - Sunlight as disinfectant. PMID- 2563447 TI - Salmonella enteritidis phage type 4 and hens' eggs. PMID- 2563449 TI - Atlantoaxial instability in Down syndrome. PMID- 2563448 TI - Assessment of fetal renal reserve in low level obstructive uropathy. PMID- 2563450 TI - Patent foramen ovale or left atrial thrombi in unexplained arterial embolism. PMID- 2563451 TI - Treatment of cocaine abuse with mazindol. PMID- 2563452 TI - Maternal allele loss in Wilms' tumour. PMID- 2563453 TI - Kidney for sale by live donor. PMID- 2563454 TI - Hypoxic-reperfusion injury in the inflamed human joint. AB - A series of experiments showed that, on exercise of the inflamed human knee, intra-articular pressure rises above synovial capillary perfusion pressure, causing intra-articular hypoxia; and that, on cessation of exercise, there is oxidative damage to lipids and IgG within the joint. These findings are consistent with the hypothesis that persistence of synovial inflammation can be due to exercise-induced hypoxic-reperfusion injury mediated by reactive oxygen species. PMID- 2563455 TI - Insulin deficiency in non-insulin-dependent diabetes. AB - A highly specific two-site immunoradiometric assay for insulin was used to measure the plasma insulin response to 75 g glucose administered orally to 49 patients with non-insulin-dependent diabetes (NIDDM). The plasma insulin concentration 30 min after glucose ingestion was lower in the diabetic patients than in matched controls for both non-obese (11-83 pmol/l vs 136-297 pmol/l, p less than 0.01) and obese subjects (23-119 pmol/l vs 137-378 pmol/l, p less than 0.01). By means of another two-site immunoradiometric assay, the basal intact proinsulin level was found to be higher in the NIDDM patients than in the controls for both non-obese (7.1 [SEM 1.2] pmol/l vs 2.4 [0.4] pmol/l, p less than 0.01) and obese subjects (14.4 [2.2] pmol/l vs 5.9 [1.9] pmol/l, p less than 0.01). The basal level of 32-33 split proinsulin was also raised in NIDDM. Previous failure to show clear separation between normal and NIDDM insulin responses was probably due to the high concentrations of proinsulin-like molecules in the plasma of NIDDM patients. These substances cross-react as insulin in most, if not all, insulin radioimmunoassays but have very little biological insulin-like activity. It is therefore now possible and necessary to designate most NIDDM patients as insulin deficient. PMID- 2563456 TI - Effect of breast-feeding on immune response to BCG vaccination. AB - The effect on BCG immunisation of feeding either formula or breast milk was assessed in Canadian Cree infants who were vaccinated either at birth or after 1 month of age. The response to BCG was measured in terms of lymphocyte blastogenesis stimulated by purified protein derivative of Mycobacterium tuberculosis. Breast-feeding significantly enhanced cell-mediated immune response to BCG vaccine given at birth, but had no significant effect if vaccine was given after 1 month. These findings were not related to maternal history of tuberculosis or BCG vaccination, and the feeding method did not influence lymphocyte stimulation by candida or streptococcal antigens. PMID- 2563457 TI - Assessment of clinical criteria for identification of severe acute lower respiratory tract infections in children. AB - 222 acute lower respiratory tract infections (LRI), as defined by the World Health Organisation, were identified during one year's surveillance of a cohort of 500 Gambian children aged 0 to 4 years. Symptoms and signs at presentation were related to radiological evidence of lobar consolidation, indicating severe LRI. In infants, a fever of greater than 38.5 degrees C, refusal to breast-feed, or the presence of vomiting were the best predictors of severe LRI. In children aged 1 to 4 years, a fever of greater than 38.5 degrees C or a respiratory rate greater than 60/min were the most accurate clinical signs for severe LRI. Chest indrawing did not discriminate severe LRI. These community-based findings differ from results of hospital-based studies. PMID- 2563458 TI - Radioimmunoimaging for diagnosis of bone marrow involvement in breast cancer and malignant lymphoma. AB - Granulopoietic bone-marrow was scintigraphically imaged in 15 patients with carcinoma of the breast and known skeletal metastases, 10 patients with malignant lymphomas, and 15 controls without suspected malignant disease, with a technetium 99m labelled murine monoclonal IgG1 antibody directed against nonspecific cross reacting antigen (NCA-95) and carcinoembryonic antigen. Immunoscintigraphy revealed more lesions than did bone scanning in both patient groups. This method offers a sensitive, cost-effective, and easy-to-perform whole body technique for evaluating metastatic spread. PMID- 2563460 TI - Breath of life. PMID- 2563459 TI - Cytomegalovirus infection in allograft recipients. PMID- 2563461 TI - Dr Koop's diet. PMID- 2563462 TI - Insulin pen: mightier than syringe? PMID- 2563463 TI - Interleukin-2: sunrise for immunotherapy? PMID- 2563464 TI - Neuropathology of the brain in HIV infection. AB - The brains of 26 patients infected with the human immunodeficiency virus (HIV) were examined post mortem. All patients were men, aged 20-67 years (mean 38.8). 13 (50%) were homosexual, 3 (12%) were bisexual, 8 (31%) were haemophiliac, 1 was both an intravenous drug addict and homosexual, and 1 denied belonging to any risk group. Only 3 (12%) brains were normal, whereas 23 (88%) showed abnormalities that varied in severity and complexity. 11 were affected by more than one disease. In addition to neoplasms, opportunistic infections, and vascular lesions, 6 cases of HIV encephalitis were found, characterised by multinucleate giant cells which indicate the presence of HIV. Microglial macrophage nodules (nodular encephalitis) occurred in 5 cases. Cerebral pathology differed between risk groups: all 6 patients with HIV encephalitis were homosexuals, whereas vascular lesions were more common in haemophiliacs. These observations have fundamental implications for clinical practice and indicate the importance of neuropathological examination in AIDS. PMID- 2563465 TI - Screening for otitis media with effusion in preschool children. AB - 1439 Dutch children were included in a randomised trial to evaluate the efficacy of preschool screening for otitis media with effusion (OME) by 3-monthly tympanometry. Children with bilateral OME on two consecutive occasions were referred for further investigation and then, if parents gave their consent, allocated at random to treatment or non-treatment groups. The effect of childhood screening for OME and subsequent treatment was evaluated by assessment of language performance; no benefit was found, mainly because of the large degree of spontaneous recovery. PMID- 2563466 TI - Isothiazolinone preservative: cause of a continuing epidemic of cosmetic dermatitis. AB - A preservative system for cosmetics and toiletries containing as active ingredients a mixture of methylisothiazolinone and methylchloroisothiazolinone (1.5%) is an important cause of cosmetic allergy in many European countries. Most cases have been caused by products of the "leave-on" variety, such as moisturising creams. The use of isothiazolinone preservative in such products should be abandoned. More critical evaluation of its sensitising potential before marketing might have prevented the continuing epidemic of allergic cosmetic dermatitis due to this preservative. New chemicals should undergo extensive toxicological evaluation before their use in cosmetics is allowed. Ingredient labelling should be made a legal requirement. PMID- 2563467 TI - Lochgoilhead fever: outbreak of non-pneumonic legionellosis due to Legionella micdadei. AB - Analysis of case histories from 187 people who had visited a hotel and leisure complex in Lochgoilhead, a village on the west coast of Scotland, indicated that 170 had had an acute illness characterised by headache, fatigue, arthralgia, myalgia, cough, and breathlessness. These symptoms were consistent with Pontiac fever-like illness. Legionella micdadei was isolated from the leisure complex whirlpool spa at the time that 60 of 72 individuals with symptoms seroconverted to L micdadei antigen. This outbreak is thought to be the first of a Pontiac fever-like illness ascribed to L micdadei and the first large-scale outbreak of its kind to have occurred outside North America. Whirlpool spas can be a major reservoir of legionella organisms; they must therefore be properly maintained and operated to prevent outbreaks of infection. PMID- 2563468 TI - Gender, lipoproteins, diet, and cardiovascular risk. Sauce for the goose may not be sauce for the gander. PMID- 2563469 TI - Listeria in food: a veterinary perspective. PMID- 2563471 TI - Cheese-borne listeria meningitis in immunocompetent patient. PMID- 2563470 TI - Listeriosis in pregnancy. PMID- 2563472 TI - Binding assay to predict response to H2-antagonists. PMID- 2563473 TI - Diffuse Lewy body disease. PMID- 2563474 TI - What makes microbes stick? PMID- 2563476 TI - Atrial fibrillation and antithrombotic prophylaxis: a prospective meta-analysis. PMID- 2563475 TI - Peripheral laser angioplasty with the pulsed dye laser and ball-tipped optical fibres. PMID- 2563477 TI - Correcting total serum calcium. PMID- 2563478 TI - Pyelonephritic Escherichia coli strains as intestinal pathogens in two newborn infants. PMID- 2563479 TI - Gender and survival in neuroblastoma. PMID- 2563480 TI - Treatment of hyponatraemia secondary to water overload. PMID- 2563481 TI - CD4+ T-cell damage to islet beta cells. PMID- 2563482 TI - The white-coat effect and self-recording of blood pressure. PMID- 2563483 TI - Bus-door accidents. PMID- 2563484 TI - Destitution at the festive season. PMID- 2563485 TI - Rheumatoid arthritis and coeliac disease. PMID- 2563486 TI - Systemic capillary leak syndrome. PMID- 2563487 TI - Infective cause of childhood leukaemia. PMID- 2563488 TI - Short stature and malnutrition in cystic fibrosis. PMID- 2563489 TI - Rapid diagnosis of cytomegalovirus infection in renal transplant recipients. PMID- 2563491 TI - Breastfeeding and HIV. PMID- 2563490 TI - Clinical reactivation of hepatitis B in anti-HBs-positive patients with AIDS. PMID- 2563492 TI - Duodenal ulcer and Campylobacter pylori. PMID- 2563493 TI - Transdermal nicotine and placebo. PMID- 2563494 TI - Chorionic villus sampling or amniocentesis. PMID- 2563495 TI - Is cervical laser therapy painful? PMID- 2563496 TI - Skin test for penicillin hypersensitivity. PMID- 2563497 TI - Alcohol and the U-shaped curve. PMID- 2563498 TI - Single injection for treatment of community-acquired pneumonia. PMID- 2563499 TI - Confidence intervals and trial sizes. PMID- 2563500 TI - Cisapride and cystic fibrosis. PMID- 2563501 TI - Multiple sclerosis or blood-brain barrier disease. PMID- 2563502 TI - Chinese medicine interfering with digoxin immunoassays. PMID- 2563503 TI - Correction of corticosteroid-induced osteoporosis by percutaneous hormone implants. PMID- 2563504 TI - Fluconazole and candidosis. PMID- 2563505 TI - Incompetent adults and consent to treatment. PMID- 2563506 TI - Migration and geographic variations in ischaemic heart disease in Great Britain. AB - The British Regional Heart Study seeks to explain the geographic variations in cardiovascular disease in Great Britain. A strong geographic gradient in the risk of a major ischaemic heart disease (IHD) event was found in 7735 middle-aged men who were followed up for 6.5 years. Regardless of where they were born, men examined in Scotland experienced the highest IHD risk, while those examined in the South of England had the lowest. The place of examination (ie, residence) was a more important determinant of the risk of a major IHD event than the place of birth. It seems unlikely that the geographic differences in IHD risk among middle aged British men can be directly explained by their genetic inheritance or by their prenatal and postnatal diet. PMID- 2563507 TI - Anorectal varices, haemorrhoids, and portal hypertension. AB - In a prospective study of 100 consecutive patients with cirrhosis, 44% had anorectal varices. The prevalence of anorectal varices rose with progression of portal hypertension; it was 19% in cirrhotic patients without portal hypertension compared with 59% in those who had bled from oesophageal varices. There was no evidence that endoscopic sclerotherapy directly increased the prevalence of anorectal varices. Haemorrhoids occurred independently of anorectal varices and their presence was unrelated to the degree of portal hypertension. These data provide further evidence that haemorrhoids and anorectal varices are separate and distinct entities. However, both can bleed and careful examination is essential to prevent misdiagnosis and inappropriate treatment. PMID- 2563508 TI - Predisposing locus for Alzheimer's disease on chromosome 21. AB - Linkage between Alzheimer's disease and markers on the long arm of chromosome 21 was investigated in six families affected by disease of early onset. Linkage was confirmed and the disease locus shown to be centromeric to the locus D21S1/S11 on the long arm of the chromosome. It is argued that the data are consistent with the notion that all patients with Alzheimer's disease of genetic aetiology have a predisposing locus on chromosome 21. PMID- 2563509 TI - Antibodies as a barrier to kidney transplantation. PMID- 2563510 TI - Surgical treatment of hypertrophic obstructive cardiomyopathy. PMID- 2563511 TI - Microbiology at the crossroads. PMID- 2563512 TI - Testicular descent revisited. PMID- 2563513 TI - Haemolytic disease of the newborn. PMID- 2563514 TI - Chorioamnionitis: cause or effect? PMID- 2563515 TI - 348 cases of suspected neonatal alloimmune thrombocytopenia. AB - Serological and clinical data were collected in 348 cases of suspected neonatal alloimmune thrombocytopenia (NAT). Of the 144 mothers who were Zwa-negative, 107 had Zwa antibodies--alone (94); with HLA antibodies (12); or with Bra antibodies (1). Antibodies were detected in 12 of the 204 Zwa-positive mothers as follows: anti-Bra (9), anti-Zwb (1), anti-Baka with HLA antibody (1), and blood group B isoagglutinins (1). The frequency of NAT due to Bra incompatibility (19%) was second to Zwa (78%). Zwa-NAT was clinically the more severe (14% had intracranial haemorrhages) and responded well to either maternal platelet transfusions or intravenous IgG. In Bra-NAT intracranial haemorrhages were not observed and most children recovered without specific therapy. PMID- 2563516 TI - Misuse of verapamil in pre-excited atrial fibrillation. AB - Of 18 patients who attended accident and emergency departments with pre-excited atrial fibrillation, 10 were inappropriately treated with intravenous verapamil. The reason for the inappropriate treatment was misdiagnosis of the arrhythmia, although diagnostic electrocardiograms were available for all patients: in only 3 of the 18 patients was the correct diagnosis made before intervention. Misdiagnosis occurred because of failure to consider pre-excitation as a possible diagnosis, rather than bias towards a single alternative arrhythmia. The use of intravenous verapamil was associated with deterioration in the clinical condition of 6 patients and continued arrhythmia in all 10. PMID- 2563517 TI - Protective effect of rear-seat restraints during car collisions. AB - The nature of injuries to 2684 car occupants involved in 1055 car accidents were analysed. Less than 1% front-seat occupants were children, compared with 25% of rear-seat passengers. Nearly all (97%) rear-seat passengers were unrestrained. Type of impact was generally similar for front-seat as for back-seat occupants, except for rollover impacts, which were commoner among rear-seat passengers. Injury severity distribution was similar for front-seat as for rear-seat occupants. Except for minor-to-moderate neck injuries, which were the result of deceleration, most injuries to rear-seat passengers were due to contact with the front seat, with glazing materials, or with other parts of the car. The use of car restraints by rear-seat passengers should reduce the incidence and severity of injuries. PMID- 2563518 TI - Blood cholesterol: is population screening warranted in the UK? PMID- 2563519 TI - Tamoxifen and the uterus and endometrium. PMID- 2563520 TI - Oral contraception and genital-tract malignancy. PMID- 2563521 TI - Cervical cancer deaths in young women. PMID- 2563522 TI - Caffeinated beverages and decreased fertility. PMID- 2563523 TI - Trends in unexpected infant deaths in Sheffield. PMID- 2563524 TI - Infective cause of childhood leukaemia. PMID- 2563525 TI - Vascular access for haemodialysis. PMID- 2563526 TI - Nebulised budesonide in severe childhood asthma. PMID- 2563527 TI - Interpretation of 99mTc-HMPAO washout after stroke. PMID- 2563528 TI - Regular intake of analgesic mixtures and risk of end-stage renal failure. PMID- 2563529 TI - Valproate, spina bifida, and birth defect registries. PMID- 2563530 TI - Mianserin. PMID- 2563531 TI - Gluten IgA antibodies and coeliac disease. PMID- 2563532 TI - Diarrhoea due to 5-aminosalicylic acid in breast milk. PMID- 2563533 TI - Listeria monocytogenes and chilled foods. PMID- 2563534 TI - Continuous positive airway pressure. PMID- 2563536 TI - Seizure after zidovudine overdose. PMID- 2563535 TI - Duodenal ulcer relapse after eradication of Campylobacter pylori. PMID- 2563537 TI - HIV-2 in west Africa in 1966. PMID- 2563538 TI - Overdosage of zidovudine. PMID- 2563539 TI - Biliary pseudolithiasis and drugs. PMID- 2563540 TI - Ultrasonography and non-quantitative chorionic gonadotropin assay in diagnosis of subacute ectopic pregnancy. PMID- 2563541 TI - Plastic or paraffin? PMID- 2563542 TI - Morbidity and preterm delivery: importance of 100% follow-up. PMID- 2563544 TI - Endoscopic biopsy forceps and transfer of tissue between cases. PMID- 2563543 TI - Teachers, stress, and mortality. PMID- 2563545 TI - Epileptic seizures and smother-proof pillows. PMID- 2563546 TI - Histiocytic medullary reticulosis: a lethal form of primary EBV infection in young children in Taiwan. PMID- 2563547 TI - Postictal SPET in epilepsy. PMID- 2563548 TI - Staphylococcus lugdunensis endocarditis. PMID- 2563549 TI - Cysticercosis causing single, small CT lesions in Indian patients with seizures. PMID- 2563550 TI - Anaesthetic failure due to faulty agents. PMID- 2563551 TI - Follow-up of NANB hepatitis outbreak in plasmapheresis unit. PMID- 2563552 TI - Alpha-interferon and post-transfusion NANB hepatitis. PMID- 2563553 TI - Efficacy of flumazenil in COPD patient with therapeutic diazepam levels. PMID- 2563554 TI - Arthropod venom and bone marrow aplasia/rhabdomyolysis. PMID- 2563555 TI - Local anaesthesia for cataract surgery. PMID- 2563556 TI - Sex difference in D7S8 marker allele distribution in adult cystic fibrosis patients. PMID- 2563557 TI - Identical mitochondrial DNA deletion in blood and muscle. PMID- 2563558 TI - Anaphylactic reactions to teniposide. PMID- 2563559 TI - Motoneuron disease and exposure to solvents. PMID- 2563560 TI - Access to medical reports. PMID- 2563561 TI - Licensing authority's use of drug manufacturer's confidential data. PMID- 2563563 TI - AIDS in the UK and world wide. PMID- 2563562 TI - Consensus statements on HIV transmission. PMID- 2563564 TI - Immunoadhesins for HIV infection. PMID- 2563565 TI - [Immunoreactive somatostatin in the peripheral olfactory system and nasal mucosa of the human]. AB - Immunohistochemically, the cyclic retradecapeptide somatostatin was demonstrated in periglomerular cells and fibres of the glomerular layer (Fig. 3) of the human olfactory bulb as well as in the superficial granule layer and fibres (Fig. 2) of the olfactory tract. Additionally, somatostatin was found in endothelial cells of subepithelial arterioles and capillaries of the regio olfactoria (Fig. 4a, b) and superior turbinate. We assume that somatostatin has a paracrine transmitter function in the peripheral olfactory system and a regulative function on the blood flow of the superior nasal mucosa. PMID- 2563566 TI - Attenuation of ethanol intoxication by alpha-2 adrenoceptor antagonists. AB - The interaction of a highly potent and selective alpha-2 adrenoceptor antagonist, atipamezole with ethanol was investigated in tests assessing a number of ethanol's behavioral effects. Atipamezole antagonized ethanol's effects on directed exploration in a holeboard test, reduced observer-rated intoxication and also reduced the duration of loss of righting reflex caused by ethanol. Similar effects were produced by another alpha-2 adrenoceptor antagonist idazoxan. The magnitude of the effects was comparable to that produced in the same animal models by the imidazodiazepine Ro 15-4513, which antagonizes ethanol by an action at central benzodiazepine receptors. Whereas Ro 15-4513 possesses marked behavioral effects on its own, atipamezole is comparatively inactive in all paradigms so far tested. The data suggest that alpha-2 adrenoceptors can play an important role in modulating the intoxicating effects of ethanol. PMID- 2563567 TI - N-terminal basic amino acids are not required for translocation and processing of preproparathyroid hormone. AB - An N-terminal deletion mutant of preproparathyroid hormone that contains a single basic amino acid, lysine, in the N-terminal domain of the signal peptide is translocated across the endoplasmic reticulum membrane similarly to intact preproparathyroid hormone. To examine the function of charged residues preceeding the hydrophobic core, the lysine was replaced by an uncharged (methionine) or negatively charged (glutamic acid) amino acid. The translocational activity of the mutant signal peptides was assayed in a reticulocyte lysate system containing chicken oviduct microsomal membranes. Altering the net charge of the N-terminal domain did not abolish signal sequence activity, although the efficiency of translocation was decreased for the mutant with a glutamic acid substitution. Posttranslational, ribosome independent, translocation was observed for all the mutants tested, with the same dependence on N-terminal charge but with much lower efficiency than cotranslational translocation. These studies show that the presence of basic amino acids in the N-terminal domain of a eukaryotic signal sequence is not required for its activity. PMID- 2563569 TI - In vivo binding pattern of a trans-regulator of homoeotic genes in Drosophila melanogaster. AB - The specification and maintenance of the metameric pattern in Drosophila melanogaster is regulated by complicated gene interactions. The differential expression of the homoeotic genes of the Antennapedia complex (ANT-C) and bithorax complex (BX-C), which determine segmental identities, is partly controlled by cross-regulatory interactions of loci within the two clusters and partly by trans-acting factors located outside the two complexes. One of the trans-regulatory genes, Polycomb (Pc), acts as a repressor of the ANT-C and BX-C. Mutations of Polycomb result in a complete depression of the homoeotic genes, leading to abdominal transformations of all body segments. Polycomb is part of a large class of trans-regulatory genes (Pc-group), estimated to comprise up to 40 loci. We have raised antibodies against the Polycomb protein, and, using an improved immunostaining technique, showed that the Polycomb protein binds to 60 discrete sites along the polytene chromosomes of salivary glands. These sites comprise the ANT-C and the BX-C as well as several locations of Pc-group genes. This is the first clear evidence for a direct interaction of Polycomb with homoeotic loci and other Pc-group genes. PMID- 2563568 TI - Transgenic mice overexpressing the mouse homoeobox-containing gene Hox-1.4 exhibit abnormal gut development. AB - The mouse homoeobox-containing genes exhibit temporally and spatially specific patterns of expression in embryonic and adult tissues and are thought to be important in regulation of development and cellular differentiation, perhaps by mechanisms analogous to homoeotic genes in Drosophila melanogaster. There has been no direct demonstration that expression of these mammalian genes can affect developmental processes, however. Hox-1.4, like other mouse homoeobox-containing genes, has been shown to be expressed in specific regions of the mid-gestation embryo, but is unique in that its highest level of expression in the adult animal is restricted to developing male germ cells. We have introduced a construct carrying the mouse Hox-1.4 gene into the germ line of mice to begin to identify the cis-acting elements required for proper expression and to assess the consequences of increasing Hox-1.4 gene expression. The construct was designed to produce normal Hox-1.4 protein from transcripts that are distinguishable from the products of the endogenous gene. The integrated transgene seemed to exhibit the appropriate tissue specificity of expression, but transcript levels were elevated in certain tissues, particularly the embryonic gut. This overexpression correlated with changes in the normal developmental program of the gut, resulting in an inherited abnormal phenotype known as megacolon. PMID- 2563570 TI - Using caged neurotransmitters. AB - The development of photolabile caged neurotransmitters has made possible the study of the split-second kinetics of receptor-ligand interactions. An instrument has now been developed for activating the neurotransmitter and measuring the neuron's response. PMID- 2563571 TI - The possible role of new beta-adrenoceptor agonists in the treatment of obesity. PMID- 2563572 TI - Abnormal serum gamma-glutamyltranspeptidase in alcoholics. Clues to its explanation. AB - In order to investigate the reason for the elevation of serum gamma glutamyltranspeptidase (GGT) after chronic alcohol consumption, the activity of this enzyme, together with the activities of aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase in serum (parameters of liver cell damage) and the excretion of D-glucaric acid (D-GA) in urine (parameter of microsomal enzymatic induction) were determined in 72 chronic alcoholics. Of these, 32 had no significant liver disease (1st group) and 40 had an overt liver disease varying from fatty liver to liver cirrhosis (2nd group). The GGT was elevated in only 62% of the patients of the first group, but in 95% of the second group. Of the latter group, patients with cirrhosis had significantly higher GGT mean levels than the patients with fatty liver. On the other hand, increased D-GA excretion was only found in 23% of the group 1 patients and in 44% of the group 2 patients. Moreover, in all patients there was a significant correlation between the values of GGT and aspartate aminotransferase, but not between GGT and D-GA. From these results, the GGT increase in chronic alcoholics, would seem to be better related to cellular damage than to enzymatic induction assessed on the basis of D-GA urinary excretion. PMID- 2563573 TI - Detection of the early steps of cadmium nephropathy--comparison of light- and electron-microscopical patterns with the urinary enzymes excretion. An experimental study. AB - Few data are yet available comparing the histological patterns of cadmium nephropathy with the values of urinary enzyme excretions, useful indexes of renal tubular damage. 40 Wistar rats, divided into four groups (A-D), were intoxicated with cadmium chloride (CdCl2) at 16 ppm in drinking water for 4, 16, 40 and 60 weeks, respectively. At the end of each period all the intoxicated rats and 5 controls were assessed for creatinine clearance, fractional excretion of gamma glutamyltransferase (UfrGGT) and alpha-glucosidase (UfrAGL), indexes of anatomical tubular damage, and for fractional clearance of lysozyme (CfrLys), index of functional tubular damage. Thereafter, the rats were sacrificed and their kidneys examined with light and electron microscopy. Control rats and group A and B rats did not show any histological impairment. A widespread vesiculation of proximal tubular cells with mitochondrial and lysosomal alterations was found in the group C rats and was more evident in group D. The brush border never showed any damage in all groups in accordance with the finding of a normal excretion pattern of UfrGGT, an enzyme situated in this structure. The UfrAGL was increased only in group D rats (p less than 0.025), who showed the most severe anatomical damages. The CfrLys, an index of tubular function, was elevated in group C and D rats (p less than 0.02 and p less than 0.002, respectively). It was possible to detect the initial renal tubular damage. PMID- 2563574 TI - Renal tubular dysfunction in pancreatic cancer and chronic pancreatitis. AB - Urinary excretion of alpha-glucosidase (AGL), gamma-glutamyltransferase (GGT) and ribonuclease (RNase), and serum amylase and immunoreactive trypsin (IRT) were determined in 38 control subjects, 48 patients with pancreatic cancer, 77 with chronic pancreatitis and 47 with extrapancreatic diseases in order to ascertain the presence of a renal tubular damage and to investigate its etiology. A significantly increased frequency of pathological results for all urinary enzymes was documented in the various groups of patients as compared to controls. Significant correlations were detected among AGL, GGT and RNase. Considering the subjects as a whole, GGT and RNase excretions correlated with serum IRT and amylase; the two urinary enzymes were found to be higher when jaundice was present. In chronic pancreatic disease enzymuria was related to increased serum pancreatic enzymes; in extrapancreatic diseases it was associated to hyperbilirubinemia. The vast majority of patients with pancreatic cancer and elevated urinary enzymes presented hepatic metastases and/or jaundice. We can conclude that an anatomical and functional tubular impairment is detectable in some patients with chronic pancreatic and extrapancreatic diseases. Tubular damage seems to least in part to be related to pancreatic inflammation and necrosis in chronic pancreatic disease, while jaundice may be found to play an important role in diseases of the hepatobiliary tract. In pancreatic cancer, liver dysfunction (presence of liver metastases and/or extrahepatic cholestasis) also appears to be involved in altering tubular cells. PMID- 2563575 TI - Plasma concentration of atrial natriuretic peptide in different phases of Korean hemorrhagic fever. AB - Korean hemorrhagic fever (KHF) is an epidemic viral disease characterized by high fever, hemorrhagic tendency and renal failure, and by hemorrhages of right atrium and renal medulla as well as necrosis of anterior hypophysis. Plasma immunoreactive atrial natriuretic peptide (irANP) levels of 15 patients in the oliguric phase was 94.8 +/- 8.4 pg/ml (mean +/- SEM), 80% higher than of the normal control group (53.0 +/- 4.7 pg/ml; n = 28). In the diuretic phase it declined to 63.7 +/- 5.3 pg/ml (n = 26). Plasma renin activity (PRA) in the oliguric phase was 19.0 +/- 1.3 ng AI/ml/h, and in the diuretic phase 5.3 +/- 0.9 ng AI/ml/h, significantly higher than the control value (2.5 +/- 0.1 ng AI/ml/h). Elevations of irANP and PRA were not correlated in each group. Also systemic blood pressure as well as heart beats were significantly increased in the oliguric phase. These findings suggest that the increased irANP may have resulted from increased circulatory volume and that the ANP secretory process may not be affected by the disease. PMID- 2563576 TI - [A case of spider bite in Hungary]. AB - The authors introduce their newest case of spider-bite. They demonstrate the consequences of two types of spider bite based on referencial datas. PMID- 2563577 TI - Expression pattern of the neu (NGL) gene-encoded growth factor receptor protein (p185neu) in normal and transformed epithelial tissues of the digestive tract. AB - The neu gene (also called NGL, erbB-2, and HER-2) encodes a 185-190 kDa transmembrane glycoprotein, p185neu, which has tyrosine-specific kinase activity and is homologous to but distinct from the epidermal growth factor receptor. The normal expression of neu mRNA and protein has been demonstrated in epithelial tissues of adult animals. Also, activation of the neu oncogene has been implicated in a variety of human adenocarcinomas. In the present study, we examined the expression of the p185neu protein in normal and transformed digestive tract tissues and in a panel of digestive tract-derived cell lines. By immunohistochemistry, strong reactivity was observed in the mucosal epithelium of the stomach, small intestine, and colon of both rodents and humans. In the small intestine, there was prominent p185neu expression by mucosal epithelium of the villus, with little or no staining in the crypts. Prominent expression was observed in the liver parenchyma, the endocrine and exocrine portions of the pancreas, and in the salivary gland. Immunoreactive p185neu was also demonstrated in fetal human intestinal epithelium. Tissue sections of selected benign and malignant colonic neoplasms were also examined. Immunoreactivity was consistently greater in adenomatous polyps than in adjacent normal colonic epithelium or areas showing malignant degeneration. By radioimmunoprecipitation, there was decreased expression in cell lines derived from more anaplastic colonic tumors. The p185neu protein is expressed widely in normal and transformed epithelial tissues of the digestive tract of the adult rat and human. This finding suggests that p185neu, a putative growth factor receptor, may play a role in the regulation of normal growth and function or in the malignant transformation of these cells. PMID- 2563578 TI - Rapid carrier and prenatal diagnosis of Duchenne and Becker muscular dystrophy. PMID- 2563579 TI - Four RFLPs of the human insulin receptor gene: PstI, KpnI, RsaI (2 RFLPs). PMID- 2563580 TI - A new EcoRI polymorphism for the insulin receptor gene. PMID- 2563581 TI - An allelic polymorphism of the angiotensinogen gene in mice. PMID- 2563582 TI - Isolation and mapping of a polymorphic DNA sequence (pB3.811) on chromosome 12 [D12S33]. PMID- 2563583 TI - Two polymorphic TaqI sites at the human NGFR locus (17q12----17q22). PMID- 2563584 TI - A moderately frequent HindIII polymorphism at the human NGFR locus (17q12--- 17q22). PMID- 2563585 TI - A rare PvuII RFLP at the CRYB1 locus (17q11.2----17q12). PMID- 2563586 TI - A two allele DNA polymorphism of the human phenylethanolamine N-methyltransferase (hPNMT) gene identified by HGIA I. PMID- 2563587 TI - [Asymptomatic Caroli's disease limited to the left hepatic lobe. A cause of isolated increase of gammaglutamyltranspeptidase?]. PMID- 2563588 TI - [Acute pancreatitis during treatment with salazosulfapyridine]. PMID- 2563589 TI - Major histocompatibility complex class I-specific cytolytic T cells, derived from gld mice, lacking Thy-1, CD4, and CD8. AB - Thy-1 is a major cell surface molecule expressed on murine thymocytes and peripheral T cells. Its physiological function is unknown, but in vitro studies suggest that Thy-1 may transmit activation signals to T cells and may play a role in the growth and/or differentiation of thymocytes [Kroczek, R. A., Gunter, K. C., Seligmann, B. & Shevach, E. M. (1986) J. Immunol. 136, 4379-4384; Kroczek, R. A., Gunter, K. C., Germain, R. N. & Shevach, E. M. (1986) Nature (London) 322, 181-184]. However, not all mouse thymocytes are Thy-1+ [Scollay, R., Wilson, A., D'Amico, A., Kelly, K., Egerton, M., Pearse, M., Wu, L. & Shortman, K. (1988) Immunol. Rev. 104, 81-120]. In addition, C3H-gld/gld mice accumulate large numbers of Thy-1- (and Thy-1+) T-cell antigen receptor-positive CD8- CD4- (double negative) T cells in peripheral lymphoid organs. Our previous studies of these Thy-1- and Thy-1+ double negatives suggested that lack of Thy-1 expression correlated with diminished capacity to respond to T-cell stimuli. In this report, we describe a Thy-1- alpha/beta T-cell receptor-positive major histocompatibility complex-specific cytotoxic T-cell clone derived from C3H-gld/gld lymph node residing cells. The data show that, at least in this system, Thy-1 (and CD8/CD4) expression is not required for growth, cytolytic activity, or expression of functional T-cell receptor complexes in vitro and raise the possibility that Thy 1 expression may not be obligatory in vivo for development of cytotoxic T lymphocyte precursors in gld mice. PMID- 2563590 TI - Partial purification and characterization of a membrane-derived factor regulating neurotransmitter phenotypic expression. AB - Cell membrane contact induces the de novo expression of choline O acetyltransferase (CAT; acetyl-CoA: choline O-acetyltransferase, EC 2.3.1.6) activity in cultures of virtually pure neonatal rat dissociated sympathetic neurons. To identify molecular mechanisms underlying membrane-associated CAT induction, the responsible membrane component was characterized and partially purified. Substantial CAT-inducing activity was found in membranes from adult rat spinal cord and sensory and sympathetic ganglia. Whole brain membranes demonstrated significantly less activity. CAT induction in sympathetic neurons in response to spinal cord membranes was linear with respect to time, after an initial 6-hr lag. It was also linear with respect to concentrations of spinal cord protein from 2 to 100 micrograms per ml. CAT-inducing activity was extracted from spinal cord membranes by incubation with 100 mM NaCl and was purified approximately 5000-fold by DEAE ion-exchange and gel filtration chromatography. The active factor appears to be an extrinsic protein with an apparent molecular mass of 27 kDa. It is inactivated by trypsin and chymotrypsin but is moderately thermostable, retaining activity at 60 degrees C but not at 90 degrees C. PMID- 2563592 TI - gamma-Glutamyl transpeptidase localization in esophageal exfoliative cytology of a chinese population at risk for carcinoma. AB - gamma-Glutamyl transpeptidase (gamma-GT) localization in exfoliative esophageal specimens was evaluated in a population of 911 individuals aged 35 or older who lived in an area of China with a high incidence of esophageal squamous carcinoma. Of the total population, 24.4% was positive for gamma-GT. Graded histopathologic cellular classification revealed the following incidences: normal, 10.4% (43 of 413); hyperplasia, 22.0% (44 of 200); Grade I dysplasia, 42.4% (98 of 231); Grade II dysplasia, 46.0% (23 of 50); near-carcinoma, 100% (4 of 4); and squamous carcinoma, 77.0% (10 of 13). In these groups gamma-GT-positive cells were usually from normal esophageal squamous epithelium. A lower incidence of gamma-GT positive cells was found for dysplastic cells (dysplasia grade I, 6.5% (15 of 231); grade II, 10.0% (5 of 50). The presence of gamma-GT-positive cells in normal esophageal squamous epithelium may prove useful for identification of suspect populations; the presence in dysplastic cells may serve as a diagnostic marker for patients who will soon progress to the carcinoma stage and who should receive early treatment and careful follow-up. PMID- 2563593 TI - International Society of Radiology and International Congress of Radiology. PMID- 2563591 TI - A plasmid-encoded regulatory gene, rns, required for expression of the CS1 and CS2 adhesins of enterotoxigenic Escherichia coli. AB - To be virulent, enterotoxigenic Escherichia coli (ETEC) must produce a toxin and a pilus-like structure that mediates specific attachment to host tissue. Expression of two of these specific adherence structures, CS1 and CS2, requires the presence of a plasmid in an ETEC strain of a particular serotype and biotype. We show here that this plasmid does not contain the structural gene for a pilin protein, as previously believed. Instead we have identified a plasmid-encoded gene called rns that is required for expression of CS1 or CS2 colonization factor antigens and for adhesion. The rns gene, defined by two separately isolated insertion mutations, produces a 26-kDa protein when transcribed and translated in vitro. At the protein level the rns gene product is homologous to AraC, a positive regulator of the arabinose operon of enteric bacteria, and to RhaR and RhaS, which regulate the rhamnose operon of E. coli. The homology of the Rns protein to AraC is localized to regions that are believed to bind to DNA. Moreover, the sequence of one of these homologous regions is consistent with a DNA binding helix-turn-helix motif. The average G + C content of E. coli DNA is 50%; yet the rns gene contains only 28% G + C, suggesting that it was acquired from some other organism. PMID- 2563594 TI - Logistics of a university interventional radiology practice. PMID- 2563595 TI - Function of a bacterial activator protein that binds to transcriptional enhancers. AB - The nitrogen regulatory (NtrC) protein of enteric bacteria, which binds to sites that have the properties of transcriptional enhancers, is known to activate transcription by a form of RNA polymerase that contains the NtrA protein (sigma 54) as sigma factor (referred to as sigma 54-holoenzyme). In the presence of adenosine triphosphate, the NtrC protein catalyzes isomerization of closed recognition complexes between sigma 54-holoenzyme and the glnA promoter to open complexes in which DNA in the region of the transcription start site is locally denatured. NtrC is not required subsequently for maintenance of open complexes or initiation of transcription. PMID- 2563596 TI - Purification of growth hormone-specific transcription factor GHF-1 containing homeobox. AB - Pituitary-specific expression of the growth hormone (GH) gene is governed by a transcription factor, GHF-1, that binds to two sites within its promoter. Recently, GHF-1 was shown to be a member of the homeobox family of DNA-binding proteins. An important question is whether GHF-1 controls the expression of other pituitary specific genes, such as prolactin (Prl), expressed in closely related cell types. To this end, GHF-1 was purified from extracts of GH- and Prl expressing pituitary tumor cells and identified as a 33-kilodalton polypeptide. Although GHF-1 bound to and activated the GH promoter, it did not recognize the Prl promoter. However, at least one other factor in the same extracts, which was easily separated from GHF-1, bound to several sites within the Prl but not the GH promoter. Antibodies to GHF-1 did not react with the Prl binding activity. These results imply that the pituitary-specific expression of GH and Prl is governed by two distinct trans-acting factors. PMID- 2563597 TI - Nifedipine-induced hypotension and mesenteric ischemia. AB - We have presented a case in which nifedipine-induced hypotension in a patient receiving beta blocker and nitrate therapy precipitated mesenteric angina, leading to emergency exploratory celiotomy. Although paradoxic angina is an unusual complication of this antianginal therapy, it should be considered in any patient who has acute abdominal pain and hypotension while receiving this therapeutic regimen. PMID- 2563598 TI - Malignant islet cell tumor of the pancreas associated with high plasma calcitonin and somatostatin levels. AB - In this case report we describe a 41-year-old man, without a recognizable preexisting syndrome, who had ascites of massive proportions and died. A malignant tumor of the endocrine pancreas was identified and associated with an extremely high plasma calcitonin level. The plasma somatostatin level was also significantly elevated. Only one previous case report of a tumor with comparable secretory characteristics has been identified. PMID- 2563599 TI - Regulation of the expression of some genes for enzymes of glutathione metabolism in hepatotoxicity and hepatocarcinogenesis. AB - The reversible stage of tumor promotion, which follows the stage of initiation and precedes that of progression in multistage carcinogenesis, is a unique example of reversible toxicity in biological systems. In order to study the molecular mechanisms involved in the action of promoting agents during this stage, the regulation of the expression of genes for two enzymes of glutathione metabolism, gamma-glutamyl transpeptidase (GGT) and the placental isozyme of glutathione S-transferase (GST-P), was studied under several different conditions of promotion during multistage hepatocarcinogenesis in the rat. Promotion by phenobarbital caused an increased expression of both of these genes in altered hepatic focal lesions, although this was somewhat more variable in the case of the GGT gene. C.I. Solvent Yellow 14, an industrial dye, served as an effective promoting agent. Feeding this dye resulted in a dramatic increase in the expression of GST-P, but not that of GGT in altered hepatic foci. Factors in crude, cereal-based diets inhibited the stage of promotion by diethylnitrosamine, but enhanced promotion by phenobarbital in a synergistic manner. In contrast, at least one purified diet had the converse effect during this stage. The mRNA levels of GST-P were uniformly elevated dramatically in reversible nodules and neoplasms of rat liver that had been induced by diethylnitrosamine and phenobarbital promotion. In contrast, the level of GGT mRNA was somewhat variable, with an occasional neoplasm exhibiting almost a background level of expression of this gene. Therefore, the altered regulation of multiple genes in hepatocytes during the stage of promotion can vary with the promoting agent itself; this process may be related to the heterogeneous gene expression seen in hepatic neoplasms. A possible role for specific DNA sequences in the 5' flanking regions of such genes is considered. In addition, a cDNA clone to the mRNA of human liver GGT was isolated and sequenced. The homology of the coding sequence of the human liver GGT mRNA to that of rat kidney GGT mRNA was striking. PMID- 2563601 TI - The murine (C3H/He) epidermal Ia+ dendritic cells (Ia+DECs) and Thy-1+ dendritic cells (Thy-1+DECs) in contact hypersensitivity and aging. AB - The contact sensitivity evaluated by the ear swelling test and the dynamic changes of epidermal Ia+ dendritic cells (Ia+DECs) and Thy-1+ dendritic cells (Thy-1+DECs) were studied in trinitrochlorobenzene (TNCB) sensitized different age group C3H/He mice after challenge. A significant increase of ear swelling was observed between 6 h and 10 days of both 8-10 week (wk) and 40-48 wk groups; the ear swelling indices of 8-10 wk group were significantly higher than those of 40 48 wk group from 18 h to 5 days. A significant decrease of the densities of Ia+DECs from 18 h to 48 h, followed by a gradual increase reaching significant increase of the densities of Ia+DECs from 5 days to 21 days in both 8-10 wk and 40-48 wk groups, was observed; the densities of Thy-1+DECs significantly decreased from 18-48 h, followed by a gradual increase reaching a significant increase from 5 days to 21 days in both 8-10 wk and 40-48 wk groups. In the normal control groups, a significant decline of both Ia+DECs and Thy-1+DECs in the 40-48 wk group was observed. Results suggest that contact allergy may be diminished in aged mice. On the other hand, like Ia+DECs, Thy-1+DECs seem to be involved in the process of contact allergy. PMID- 2563600 TI - Epidermal growth factor receptor expression and growth fraction in human tumours of the nervous system. AB - 100 tumours of the human nervous system were investigated by means of immunohistochemistry in order to determine the expression of epidermal growth factor receptor (EGFr) and the proliferative activity as evaluated by demonstration of the proliferation-associated Ki-67 antigen. Epidermal growth factyr receptor immunoreactivity was present in 79% (23/29) of the high-grade malignant gliomas examined but in only 9% (2/22) of the low-grade gliomas. Besides the gliomas, EGFr-expression was detectable in smaller amounts in most (13/15) meningiomas, in one anaplastic neurinoma and in individual tumour cells of one medulloblastoma. In addition, EGFr-expression was found in 50% (6/12) of metastatic carcinomas. Seven of eight medulloblastomas, two cerebral primitive neuroectodermal tumours (PNETs), three benign neurinomas, one ganglioneuroma, one metastatic intracerebral malignant melanoma, three spinal plasmacytomas and one immunocytoma showed no detectable EGFr-expression. Our results indicate that (1) the expression of EGFr in human tumours of the nervous system depends on the histological tumour type and (2) in the glioma group is related to the grade of malignancy. A close correlation between EGFr-expression and proliferative activity as evaluated by Ki-67 staining could not, however, be established. PMID- 2563602 TI - UVB erythema inhibited by topically applied substances. AB - The photoprotective and antierythematous effects of topical corticosteroids, acetylsalicylic acid (ASA), indomethacin (IM), butylhydroxytoluene (BHT) and diphenhydramine hydrochloride (DPH) and the influence of the application time on the formation of erythema were studied in healthy volunteers. The test substances, incorporated in o/w creams, were applied to the back in large Finn Chambers 24, 4 and 1 hours before and 1 and 4 hours after ultraviolet light B (UVB) irradiation (3 x MED). The reactions were assessed both visually and using a laser Doppler flowmetry device. When applied before irradiation, ASA and IM, significantly inhibited UVB erythema. When applied after irradiation, IM, DPH and potent corticosteroids reduced UVB erythema, but ASA, BHT and hydrocortisone did not. 5% ASA had the greatest photoprotective effect when the cream was applied 4 hours or 1 hour before irradiation. The photoprotective effect was only slight when the 5% ASA cream was applied 24 hours before irradiation. PMID- 2563603 TI - Absorption of lidocaine and prilocaine after application of a eutectic mixture of local anesthetics (EMLA) on normal and diseased skin. AB - A eutectic mixture of 5% lidocaine and prilocaine was applied under occlusion for 1 or 2 hours on 25-100 cm2 areas of normal and diseased skin, and the absorption was followed by measuring the concentrations of the drugs in the draining vein and the general circulation at different time intervals after the application. The analgesic and vascular effects in the skin were also recorded. When the mixture was applied on normal skin the absorption was more rapid from the face than from the forearm. The absorption from diseased skin was faster than that from normal skin, with higher plasma concentrations, and a more rapid but shorter anesthetic effect was noted. With the doses used the plasma levels in the general circulation were 100 times lower than those associated with toxicity. The drug concentrations in the draining vein were highest after treatment of diseased skin and were 2-90 times higher than in the general circulation. The plasma concentrations of lidocaine and prilocaine ran parallel to each other, but the prilocaine level was 10-50% lower than that of lidocaine in the draining vein and 200-300% lower in the general circulation. PMID- 2563604 TI - A low-fat diet supplemented with dietary fish oil (Max-EPA) results in improvement of psoriasis and in formation of leukotriene B5. AB - Several studies have indicated that certain lipoxygenation products of arachidonic acid, particularly leukotriene B4 (LTB4), may be involved in psoriatic pathophysiology. One way of inhibiting the formation of LTB4 is to replace arachidonic acid in phospholipids with eicosapentaenoic acid. Eicosapentaenoic acid is converted into LTB5, which has a lower biologic activity than LTB4. In the present study psoriatic patients were put on a low-fat diet supplemented with dietary fish oil (Max-EPA 30 ml daily), a source of eicosapentaenoic acid, for 4 months. Twenty-six out of 30 patients with psoriasis vulgaris completed the study. Moderate or excellent improvement was observed in 58% of the patients, while mild improvement or no change was observed in 19% and 23%, respectively. The capacity of peripheral blood neutrophils to synthesize LTB4 and LTB5 in vitro was determined after stimulation with A23187. Before the study, negligible amounts of LTB5 were formed. After 1 month the average of LTB5/LTB4 ratio was 0.42. No further increase of the LTB5/LTB4 ratio was found. There existed no relationship between the clinical response and the LTB5/LTB4 ratio. The results of the present study suggest that dietary fish oil supplementation may be used in the therapy in psoriasis. However, studies defining the dose and the quality of fish oils are imperative. PMID- 2563605 TI - Selenium in whole blood and plasma is decreased in patients with moderate and severe psoriasis. AB - Concentrations of selenium in whole blood and plasma in 113 patients with moderate to severe psoriasis were compared with those in 104 healthy reference subjects. Most of the patients (85%) had had their psoriasis for at least 10 years and all had previously been treated with PUVA baths. Selenium concentrations both in whole blood and plasma were decreased both in male and female psoriasis patients. Male psoriatics 20-49 years old and women with disease of long duration (greater than 20 years) had particularly low selenium concentrations in whole blood. The lowest whole blood values were found in a subgroup of male patients with widespread disease of long duration who had also required treatment with methotrexate and/or retinoids to control their disease. The blood selenium in patients who also had psoriatic arthritis or arthralgia did not differ from those without such symptoms. PMID- 2563607 TI - Scoring atopic dermatitis: the simpler the better? AB - Two scoring methods evaluating the severity of atopic dermatitis have been compared. One was simple, quick and compatible with a busy outpatients clinic. The other was more complicated and time consuming; it took into account most of the evaluable clinical signs of disease activity in each involved site. There was a highly significant correlation between the two methods of scoring, thus validating the simplest one; furthermore, the more complicated method was less reproducible than the simplest when used by two physicians on the same patient. This suggests that a simple and feasible scoring method is meaningful in keeping records at each visit in any patient with atopic dermatitis. Such records may then be used in retrospect for the evaluation of any new therapy. PMID- 2563606 TI - A double-blind comparison of acitretin and etretinate in the treatment of severe psoriasis. Results of a Nordic multicentre study. AB - Acitretin, the free acid of etretinate, is less lipophilic and has a much shorter terminal half-life than the parent compound. The present double-blind, randomized study compared the therapeutic effectiveness and the tolerability of acitretin (n = 127) and etretinate (n = 41) in psoriasis. Patients were treated with 40 mg daily for the first 4 weeks and with an individually adjusted dose for the subsequent 8 weeks. The average daily doses of acitretin (0.54 mg/kg/day) and etretinate (0.65 mg/kg/day) were similar. The PASI (Psoriasis Area and Severity Index) scores improved in parallel in the 2 treatment groups. At the completion of the study, the PASI score improvement was 75.8% for acitretin and 70.8% for etretinate. Both acitretin and etretinate resulted in mucocutaneous side effects. Assessments of tolerability by investigators and patients showed a statistically significant difference in favour of etretinate. These results demonstrate that acitretin and etretinate have similar therapeutic effectiveness in psoriasis. Although the tolerance to acitretin was lower than to etretinate, acitretin offers the important advantage of a much shorter period of potential teratogenicity and is, therefore, to be preferred in women of childbearing potential. PMID- 2563608 TI - Skin problems from visual display units. Provocation of skin symptoms under experimental conditions. AB - Thirty patients having skin problems experienced being caused by work with visual display units (VDU) were tested double-blind with two VDUs. One VDU had strong electrostatic and electromagnetic fields and the other VDU had an identical appearance but the electrostatic field and electromagnetic fields were practically eliminated. Approximately 80% of the patients reacted with stinging or itching in the face during the 3 hours working period with 25% relative humidity in the room. No difference between the VDUs was found with regard to provoking these symptoms. At 60% relative humidity 13 patients of 19 experienced stinging or itching in the face. Those 13 that reacted were asked to come another time and were informed that the VDU was not turned on and that all electric fields that were present came from the cable to the VDU. A green cloth was put over the VDUs. This time 11 of the 13 patients reacted with stinging and itching in spite of the fact that the VDU was turned off. The present study does not indicate that electric and electromagnetic fields are of major importance in provoking subjective skin symptoms in patients experiencing skin problems when working with VDUs. PMID- 2563609 TI - Acute febrile neutrophilic dermatosis--a marker of malignancy? AB - A retrospective survey during a 2-year period disclosed 18 patients with acute febrile neutrophilic dermatosis (Sweet's syndrome). An associated lympho- or myeloproliferative malignancy was found in 6 patients. Attacks of Sweet's syndrome preceded the diagnosis of neoplasia in 4 patients (3 months to 6 years). Some differences in symptoms and signs were found in the group of patients with associated malignancy compared with the group without, that is, male predominance, mucosal symptoms, anemia, and frequent recurrence of skin symptoms. The onset of Sweet's syndrome indicates an acute infectious disease, and the patients are frequently referred to departments of internal medicine and infectious diseases. In addition, the skin lesions may mimic those which often accompany a generalized infection (erythema multiforme, erythema nodosum, vasculitis, pustular eruptions and urticaria). Since Sweet's syndrome may precede the possibly associated malignant disease, the initial diagnosis of the syndrome is important and should be made with confidence with increasing awareness of the characteristic symptoms. PMID- 2563610 TI - The complement fixing ability of anti-basement membrane zone IgG subclass of herpes gestationis and bullous pemphigoid. AB - The antibody activities and complement fixing ability of the IgG subclass antibodies of herpes gestationis (HG) and bullous pemphigoid (BP) antibodies were investigated. IgG was prepared by DEAE Affi-Gel Blue column. IgG3 and IgG 1.2.4 fractions were isolated through Protein-A Sepharose CL-4B column. In HG, neither IgG nor C3 deposits were detected at the basement membrane zone (BMZ) in the skins incubated with IgG3 fractions, although C3 deposits were detected without IgG deposits in the skins incubated with IgG 1.2.4 fractions. In BP, IgG and C3 deposits were detected in the skins incubated with IgG3 fractions, while neither IgG nor C3 deposits were detected in the skins incubated with IgG3 fractions. These findings suggest that IgG3 might not be able to fix complement at BMZ in BP and HG, although there are some reports that complement activating ability of IgG3 is the strongest among IgG subclasses. PMID- 2563611 TI - Implantation of orthopaedic devices in patients with metal allergy. AB - Patients with a contact allergy to chromium, cobalt and/or nickel, patch test verified before implantation of a metallic orthopaedic device, were followed up years later by clinical and radiographic examination as well as with epicutaneous and intracutaneous tests. Eighteen patients had been exposed to an orthopaedic implant for several years (mean 6.3 years) containing a metal to which they were allergic. None had suffered any dermatologic or orthopaedic complications attributable to the contact allergy. PMID- 2563612 TI - Serum aminoterminal propeptide of type III procollagen in progressive systemic sclerosis and localized scleroderma. AB - Sera from 31 patients with progressive systemic sclerosis (PSS), 5 patients with widespread localized scleroderma (LS), and 3 patients with lichen sclerosus et atrophicus were analyzed for aminoterminal propeptide of type III procollagen (PIIINP) using a radioimmunoassay based on human propeptide. Thirty-eight per cent of the patients with PSS had levels above normal range, including all of the 3 patients with diffuse scleroderma. The same applies to 4 of 5 patients with widespread localized LS, while PIIINP in all 3 patients with lichen sclerosus et atrophicus were within normal levels. In patients with acrosclerosis, elevated PIIINP seems to be correlated to rapid progression and extension of lesions. A significant increase in PIIINP was found in a patient following discontinuation of prednisone and cyclophosphamide, while the present investigation did not allow judgement of effects of treatment with either penicillamine or cyclosporin A. PMID- 2563613 TI - Multiple eccrine poromas arising in chronic radiation dermatitis. AB - A 70-year-old white man developed 7 eccrine poromas in an area of chronic radiation dermatitis of his right lower extremity over a period of 37 years. To our knowledge, multiple eccrine poromas unequivocally linked to chronic X-ray damage are hitherto unreported. PMID- 2563614 TI - Intermittent leukapheresis: an adjunct to low-dose chemotherapy for Sezary syndrome. AB - Eleven patients with Sezary syndrome were treated with intermittent leukapheresis in addition to low-dose chlorambucil and prednisone. The results were as good as or better than those with chemotherapy alone. We believe the combined program with continuous leukapheresis to be optimal therapy but note that intermittent treatment offers some benefit for patients. PMID- 2563615 TI - Dithranol in the treatment of inflammatory linear verrucous epidermal nevus. AB - A case of inflammatory linear verrucous epidermal nevus (ILVEN) is reported. Short contact treatment with dithranol resulted in complete relief from itching and a remarkable clearing of all linear lesions except from a small verrucous band on the shin. In patients with ILVEN it is advisable to try dithranol therapy before carrying out surgical procedures such as excision, cryotherapy, electrocautery. The prompt response to dithranol is best explained by the assumption that most of the lesions in this case of ILVEN represented true linear psoriasis. PMID- 2563616 TI - Psoriasis of the nails treated with grenz rays: a double-blind bilateral trial. AB - The effect of grenz ray therapy in the treatment of psoriatic nails was assessed in 22 patients by randomly allocating active treatment to the psoriatic nails of one hand while the other one, which received simulated therapy, served as a control. Five Gy of grenz rays were applied on 10 occasions at intervals of 1 week. There was a significantly better response to active treatment compared with the untreated control. However, the therapeutic response was moderate. It is concluded that grenz ray therapy could be useful only when the psoriatic nails are of normal thickness. PMID- 2563617 TI - 311 nm UVB lamps in the treatment of psoriasis with the Ingram regimen. AB - A new experimental fluorescent lamp emitting UVB mainly in a narrow peak around 311 nm was compared with a conventional broad band UVB lamp in the treatment of psoriasis with the Ingram regimen. In 20 patients one arm was treated with the new lamps and the other arm with conventional lamps. In 12 patients the results were same, but the new lamp was more effective in 8 patients. In another trial, 53% of 17 patients treated with the new lamp showed good results compared with 30% of 23 patients treated with conventional lamps. In conclusion, the 311 nm UVB lamp is at least as effective as the conventional broad band UVB lamps in the treatment of psoriasis with the Ingram regimen. PMID- 2563618 TI - Peptide-T in the treatment of psoriasis and psoriatic arthritis. A case report. AB - The following is a description of the improvement of a HIV-negative patient with psoriasis and monoarthritis who received treatment with Peptide-T, a synthetic octapeptide that shares a segment of the envelope glycoprotein (gp 120) of the human immunodeficiency virus. PMID- 2563619 TI - Squamous cell carcinomas in relation to cyclosporin therapy of non malignant skin disorders. AB - Two patients with psoriasis and one with pityriasis rubra pilaris developed squamous cell carcinomas in relation to cyclosporin A therapy. Dermatological patients previously treated with ultraviolet radiation and other kinds of immunosuppressive therapy may represent a special risk group with respect to cyclosporin. A related malignancies. PMID- 2563620 TI - Cantharidin treatment of molluscum contagiosum. PMID- 2563621 TI - Duration of action of vecuronium in infants and children anaesthetized without potent inhalation agents. AB - The duration of effects of vecuronium has previously been studied in paediatric patients only during inhalation anaesthesia. We therefore studied the age-related differences in duration of action after administration of 0.1 mg kg-1 of vecuronium in 66% N2O in O2, fentanyl anaesthesia without inhalation agents. Forty-nine children (2 wk-14 yr, ASA 1-2) were selected for study and divided into four groups according to age. Evoked EMG monitor (Relaxograph, Datex, Finland) was calibrated with hypothenar muscle recording. The completely restored first twitch (T1) level was used as reference for recovery calculations. In infants (age less than 1 yr) the onset time (68 s) was only 0.6-0.8 times that of older patients (P less than 0.01). In infants the duration of surgical relaxation to T1 10% (42 min) and the recovery index (21 min) were 1.7-2.9 times longer than in older patients (P less than 0.01). It took 55 min from the beginning of recovery to full restoration of T1 in infants, compared with 20-24 min in 3-15 year-old children. Interestingly, the younger the child, the more rapidly train of-four ratio recovered compared with T1 (P less than 0.01). Because of the age dependent prolongation of vecuronium relaxation and spontaneous recovery in small children, the level of relaxation should be monitored also in clinical practice. PMID- 2563622 TI - Interactions between noradrenergic and cholinergic mechanisms involved in spinal nociceptive processing. AB - Antinociceptive effects have been demonstrated after systemic and spinal administration of the adrenoceptor agonist clonidine and cholinomimetic drugs in animals and human. The present investigation was undertaken in rats to study the possible interactions between spinal noradrenergic and cholinergic mechanisms in modulating the reaction to nociceptive stimuli. Using the tail immersion test, an additive antinociceptive effect was found between intrathecal (IT) clonidine (10 micrograms) and physostigmine (15 micrograms, IT). The effect of clonidine was attenuated by atropine (15 micrograms, IT). Physostigmine (15 micrograms, IT) antinociception, which was of short duration was abolished by atropine (15 micrograms, IT) and attenuated by phentolamine (20 micrograms, IT). Neostigmine (5 micrograms, IT) produced a prolonged antinociceptive response. In animals pretreated with 6-hydroxydopamine IT, leading to a selective depletion of spinal cord noradrenaline, physostigmine (15 micrograms, IT) was ineffective in altering the nociceptive test response. Neither clonidine, nor physostigmine produced changes in latency times in the hot plate test (58 degrees C) in the doses employed. In conclusion, a clear-cut interaction exists between spinal noradrenergic and cholinergic systems for antinociception. To explain the interactions, several possible mechanisms may be considered, including cholinomimetic effects produced by clonidine, and the presence of muscarinic receptors in the dorsal horn of the spinal cord. PMID- 2563623 TI - Subdural and epidural empyemas: MR imaging. AB - The MR images of six patients with extraaxial empyemas (five subdural and four epidural) were reviewed and compared with CT scans. MR demonstrated convexity and interhemispheric collections, which were mildly hyperintense relative to CSF and hypointense relative to white matter on short TR pulse sequences and hyperintense relative to CSF and white matter on long TR pulse sequences, allowing distinction from sterile effusions and most chronic hematomas. A hypointense rim, representing displaced dura, was depicted at the interface between the lesion and brain in epidural empyemas, a feature absent in subdural empyemas. Inflammation induced parenchymal abnormalities, including edema, mass effect, and reversible cortical hyperintensity, were well depicted on MR imaging. MR was superior to CT in demonstrating the presence, nature, and extent of these lesions in all cases. Because early and accurate diagnosis will significantly improve the prognosis of these serious infections, MR is preferred to CT for patients in whom an acute intracranial infection is suspected. PMID- 2563624 TI - Effects of dilevalol on rest and supine exercise hemodynamics in mild to moderate systemic hypertension. AB - Eight mildly to moderately hypertensive subjects free of any antihypertensive medications and on a normal salt diet performed maximal supine arm exercise. Before starting the exercise, a right-sided cardiac catheterization was performed to measure hemodynamic parameters before and during exercise. All patients had normal increases in cardiac output for the level of exercise performed and the peripheral vascular resistance diminished appropriately. An increase in the right atrial and pulmonary artery wedge pressures during exercise could be explained by increased venous return. After the baseline testing, rest and exercise hemodynamics were repeated 2 hours after the administration of 400 mg of dilevalol, a new beta blocker. For the next 2 weeks the patients received 400 mg of the study drug twice a day, with repeat studies obtained thereafter. As with other beta blockers, dilevalol decreases the heart rate and cardiac output on exercise, but, in addition, it induces a decrease in the resting systemic vascular resistance. This action is similar to its isomer, labetalol. PMID- 2563625 TI - alpha-Linolenic acid and long-chain omega-3 fatty acid supplementation in three patients with omega-3 fatty acid deficiency: effect on lymphocyte function, plasma and red cell lipids, and prostanoid formation. AB - alpha-Linolenic acid deficiency is described in three patients. Observed clinical symptoms were hemorrhagic dermatitis, hemorrhagic folliculitis, skin atrophy, and scaly dermatitis. Supplementation with ethyl alpha-linolenate followed by a purified fish oil (EPA-oil) began to normalize symptoms within 10 d. The mitogenic response in isolated lymphocytes was reduced whereas the number of T lymphocytes increased significantly. Serum thromboxanes, urinary excretion of 2,3 dinor-6-keto-prostaglandin F1 alpha (PGI2-M), and bleeding time were unaffected. The results indicate that omega-3 fatty acids are essential for normal accumulation of erythrocyte omega-6 acids. The dietary intake of long-chain omega 3 acids required to obtain midnormal concentrations of omega-3 acids in plasma and erythrocyte lipids was estimated to be 350-400 mg/d (0.4% of calories), whereas the corresponding mean intake of alpha-linolenic acid was 990 mg/d (1.0% of calories). It is suggested that essential fatty acid requirement should be stated as grams or milligrams per day, similarly to other essential nutrients. PMID- 2563626 TI - Linoleic acid and linolenic acid: effect on permeability properties of cultured endothelial cell monolayers. AB - High circulating plasma levels of free fatty acids may injure endothelial cells, resulting in decreased barrier function of the vascular endothelium. The effect of media supplementation with varying concentrations of either linoleic (C18:2 omega 6) or linolenic acid (C18:3 omega 3) on albumin transfer across cultured endothelial monolayers was studied. A 24-h cell exposure to linoleic but not linolenic acid resulted in a concentration dependent and largely reversible increase in albumin transfer. Both fatty acids and in particular linolenic acid incorporated into cellular phospholipids. In contrast, only supplementation with linoleic but not linolenic acid resulted in an increased incorporation of this fatty acid into cell triglycerides. Similarly, only total cell triglyceride content increased after incubation with linoleic- but not with linolenic-enriched media. These results indicate that cellular enrichment with linoleic but not linolenic acid causes cellular perturbations that may be implicated in atherosclerosis. PMID- 2563627 TI - The efficacy and safety of home nebulizer therapy for children with asthma. AB - We evaluated the efficacy and safety of nebulized beta-agonists used in the homes of 22 children with asthma. Patients served as their own controls for the comparison of asthma-related variables between periods of 12 months before and 12 months after the initiation of home nebulizer therapy. Significant reductions in the numbers of emergency department visits, hospital admissions, and short courses of prednisone therapy occurred when home nebulizer therapy was provided. The younger the patients, the more frequently improvement occurred. After the initiation of home nebulizer therapy there was no change in the severity of illness when patients presented to the emergency department or in the number of episodes of respiratory failure. Home nebulizer therapy for children with asthma appears safe and reduces the need for hospital care and short courses of prednisone therapy. PMID- 2563628 TI - Beneficial effects of glucose polymer and an H2-receptor blocker in a patient with a proximal ileostomy. AB - A patient with an ileocolectomy and proximal ileostomy for Crohn's disease had severe diarrhea and steatorrhea. An oral electrolyte solution containing glucose polymer was shown to improve water and electrolyte absorption. Ileal contents were abnormally acidic. Therapy with an H2-receptor blocker raised ileal pH, improved the efficiency of fat absorption, and promoted a gain in body weight. PMID- 2563629 TI - Famotidine and ranitidine, but not cimetidine, cause severe, disabling headache. PMID- 2563630 TI - An autopsy of epidemiologic methods: the case of "poppers" in the early epidemic of the acquired immunodeficiency syndrome (AIDS) PMID- 2563631 TI - Analysis of DNA polymorphism haplotypes linked to the cystic fibrosis locus in North American black and Caucasian families supports the existence of multiple mutations of the cystic fibrosis gene. AB - Strong linkage disequilibrium (LD) was found between DNA marker XV2c and the cystic fibrosis (CF) locus (delta = 0.46) and between DNA marker KM19 and CF (delta = 0.67) in 157 CF and 138 normal chromosomes from U.S. Caucasians. DNA haplotypes with nine polymorphic sites were created in 54 Caucasian families. There is a strong LD between the haplotypes and the presence of the mutant CF genes. This implies that the DNA polymorphisms examined are close to the CF gene and that one mutation of the CF gene predominates in the Caucasian population. Haplotype analysis can also be used to refine estimates of CF carrier risk in Caucasians. Data for XV2c and MET markers in 16 American black patients and their families revealed a different haplotype distribution and LD pattern with the CF locus. These data suggest that racial admixture alone does not explain the occurrence of CF in American blacks and that multiple alleles of the CF gene may exist in this population. PMID- 2563632 TI - Genealogical analysis of cystic fibrosis families and chromosome 7q RFLP haplotypes in the Hutterite Brethren. AB - In the 100-year period 1880-1980 the Hutterite population increased from about 442 to 23,000 individuals in North America. There are three endogamous subdivisions in this Caucasian genetic isolate. A total of 11 cystic fibrosis (CF) families from Canada and the United States were investigated, including at least two families from each of the three subdivisions, the Dariusleut, Lehrerleut, and Schmiedeleut. A study of RFLPs for the loci D7S8, D7S23, MET, and D7S18 (also called D7S16) in the region of the CF gene in 10 families shows considerable genetic variability. There were three different extended CF gene region haplotypes on CF chromosomes (CF haplotypes), and there were 13 different extended CF gene-region haplotypes on normal chromosomes (normal haplotypes). The three CF haplotypes have different D7S23 and MET haplotypes. Parents who have the same CF haplotype are, on the average, more closely related than parents who have different haplotypes, but only within the same subdivision. A marriage node graph of 11 families illustrates the complexity of Hutterite genealogies. The frequency distribution of CF haplotypes in the Hutterite sample differs notably from those of larger agglomerates of family data from collaborative studies, with respect to D7S8, MET haplotypes, and D7S23 haplotypes. We propose that there were at least three CF carriers among the founders of the Hutterite population and that copies of a particular CF haplotype in current individuals are identical by descent. The alternative that one or more genetically distinguishable CF haplotypes resulted from recombination since the founding of the population is considered to be less likely. PMID- 2563633 TI - Phenylalanine hydroxylase gene haplotypes in Polynesians: evolutionary origins and absence of alleles associated with severe phenylketonuria. AB - A total of 630 haplotypes for the phenylalanine hydroxylase (PAH) gene locus were established in five groups of Polynesians comprising Samoans, Tongans, Cook Islanders, Maori, and Niueans. Considerable genetic continuity was demonstrated between these widely dispersed populations, since three common haplotypes (4, 1, and 7) constituted over 95% of alleles. A control group of individuals from Southeast Asia shared the same major haplotypes, 4, 1, and 7, with Polynesians. These data provide further support for the theories of genetic homogeneity and of Asian affinities of the Polynesian precursor populations. The absence of severe phenylketonuria (PKU) in both Polynesians and Southeast Asians is consistent with the lack of PAH haplotypes 2 and 3, on which the severe PKU mutants have arisen among Caucasians. PMID- 2563634 TI - A hypervariable microsatellite revealed by in vitro amplification of a dinucleotide repeat within the cardiac muscle actin gene. AB - The human genome contains approximately 50,000 copies of an interspersed repeat with the sequence (dT-dG)n, where n = approximately 10-60. In humans, (TG)n repeats have been found in several sequenced regions. Since minisatellite regions with larger repeat elements often display extensive length polymorphisms, we suspected that (TG)n repeats ("microsatellites") might also be polymorphic. Using the polymerase chain reaction to amplify a (TG)n microsatellite in the human cardiac actin gene, we detected 12 different allelic fragments in 37 unrelated individuals, 32 of whom were heterozygous. Codominant Mendelian inheritance of fragments was observed in three families with a total of 24 children. Because of the widespread distribution of (TG)n microsatellites, polymorphisms of this type may be generally abundant and present in regions where minisatellites are rare, making such microsatellite loci very useful for linkage studies in humans. PMID- 2563635 TI - Polymorphic DNA haplotypes at the LDL receptor locus. AB - Mutations in the low-density lipoprotein (LDL) receptor gene result in the autosomal dominant disorder familial hypercholesterolemia (FH). Many different LDL receptor mutations have been identified and characterized, demonstrating a high degree of allelic heterogeneity at this locus. The ability to identify mutant LDL receptor genes for prenatal diagnosis of homozygous FH or to study the role of the LDL receptor gene in polygenic hypercholesterolemia requires the use of closely linked RFLPs. In the present study we used 10 different RFLPs, including three newly described polymorphisms, to construct 123 independent haplotypes from 20 Caucasian American pedigrees. Our sample contained 31 different haplotypes varying in frequency from 0.8% to 29.3%; the five most common haplotypes account for 67.5% of the sample. The heterozygosity and PIC of each site were determined, and these values disclosed that eight of the RFLPs were substantially polymorphic. Linkage-disequilibrium analysis of the haplotype data revealed strong nonrandom associations among all 10 RFLPs, especially among those sites clustered in the 3' region of the gene. Evolutionary analysis suggests the occurrence of both mutational and recombinational events in the generation of the observed haplotypes. A strategy for haplotype analysis of the LDL receptor gene in individuals of Caucasian American descent is presented. PMID- 2563636 TI - High-dose monotherapy and combination therapy with calcium channel blockers for angina. A comprehensive review of the literature. AB - Clinical experience with the calcium channel-blocking agents has established their efficacy in the therapy of painful and silent myocardial ischemia. Questions have arisen, however, about side-effect characteristics of these medications as clinical practice has led to utilization of higher doses of individual drugs than employed in large numbers of patients in early clinical trials as well as combinations with other antianginal agents including beta blockers. A study was undertaken to examine the published literature regarding side effects associated with high-dose versus low-dose therapy with nifedipine and diltiazem and the use of these agents in combination with beta-blockers. This investigation demonstrated that utilization of high-dose diltiazem (more than 240 mg per day) as opposed to low-dose diltiazem (no more than 240 mg per day) was associated with an increased incidence of atrioventricular block, and increased peripheral vasodilatory effects. In contrast, the use of high-dose nifedipine (more than 60 mg per day) was not associated with atrioventricular block. At clinically high dosage levels, the incidence of peripheral edema was comparable for both nifedipine and diltiazem, although low-dose nifedipine resulted in a significantly greater incidence of edema compared with low-dose diltiazem. This analysis also demonstrated that bradyarrhythmia is associated with the combination of a beta-blocking agent and either low- or high-dose diltiazem, but not with nifedipine-beta-blocker combinations. Clinical experience suggests caution in the combined use of diltiazem and a beta-blocking agent because of the demonstrated additional adverse negative chronotropic and dromotropic effects. No additional adverse electrophysiologic effects have been noted for nifedipine-beta blocker combinations. The literature analysis supports and mirrors widespread clinical experience obtained since nifedipine and diltiazem were introduced. It should be noted, though, that combination therapy with calcium channel blockers and beta-blockers should be done with caution, since there have been occasional reports of congestive heart failure or exacerbation of angina with this combination. PMID- 2563637 TI - Role of dipeptidyl peptidase IV (gp 108) in passive Heymann nephritis. Use of dipeptidyl peptidase IV-deficient rats. AB - Injection of antibodies to renal tubular membrane (Fx1A) into Lewis rats induces granular deposits of IgG in glomeruli and proteinuria (passive Heymann nephritis, PHN), and similar lesions are also induced by antibody to one of the antigens in Fx1A, dipeptidyl peptidase IV (DPP IV, gp 108). In this study, the role of DPP IV in PHN was investigated using DPP IV-deficient F344 rats. The amount of DPP IV found in F344 rat kidneys was less than 0.05% of that present in Wistar rats, and injection of anti-DPP IV antibody into F344 rats did not induce proteinuria. Injection of anti-F344 Fx1A rabbit antibodies that contain no detectable anti-DPP IV antibody into Lewis or F344 rats induced PHN, characterized by granular deposits of rabbit IgG in glomeruli and massive proteinuria, although the appearance of proteinuria was delayed in comparison with that occurring in response to injection of anti-Wistar Fx1A antibodies. These results indicate that DPP IV may contribute to, but is not essential for, the induction of PHN. PMID- 2563638 TI - Pertussis toxin effects on transmitter release from perivascular nerve terminals. AB - Pertussis toxin (PT) was used to investigate the possible involvement of an inhibitory GTP-binding protein (G protein) in neuromuscular transmission at the sympathetic nerve terminal of guinea pig mesenteric artery. When the intracellular microelectrode technique was used, the amount of transmitter released was evaluated by recording the amplitude of excitatory junction potentials (EJPs) in the vascular smooth muscle (VSM) cells. EJPs were evoked by perivascular nerve stimulation with brief square pulses. The amplitude of EJPs was depressed by exogenously applied norepinephrine (NE) (greater than or equal to 10(-7) M) or histamine (greater than or equal to 10(-7) M). However, these effects of NE and histamine on EJP amplitude were almost completely blocked in PT pretreated VSM cells (2 x 10(-7) g/ml PT, 24 h at 21 degrees C). These effects of PT pretreatment were time and temperature dependent. In contrast, there was no significant change in the resting membrane potential (-70.1 +/- 2.1 mV) or input resistance (11.9 +/- 0.4 M omega) in the VSM cells after PT pretreatment. These results suggest that the effects of NE and histamine on EJP amplitude may be mediated by PT-sensitive G proteins in the presynaptic nerve terminal. PMID- 2563639 TI - Renal sympathetic nerve activity and natriuresis during water immersion in conscious dogs. AB - The role of renal sympathetic nerve activity (RSNA) in the natriuresis and diuresis induced by head-out water immersion (WI) was studied in eight conscious female dogs. The dog was instrumented chronically with a stainless steel electrode for the measurement of RSNA and two catheters for the measurements of systemic arterial (Pa) and central venous (Pv) pressures. The WI caused an immediate reduction of RSNA by 43 +/- 7% (P less than 0.05), and this low level was sustained throughout a 120-min WI under thermoneutral conditions (37 degrees C). Urine flow and sodium excretion increased by 211 +/- 54 (P less than 0.05) and 240 +/- 122% (P less than 0.05), respectively, but creatinine clearance did not change significantly during WI. A step increase in Pa (by 10 +/- 4 mmHg, P less than 0.05) and Pv (by 10.0 +/- 0.8 mmHg, P less than 0.05) was observed also during WI. In another series of studies, renal denervations were performed 2-4 wk before the experiment in six of the same dogs. Dogs with renal denervation showed no significant changes in urine flow and sodium excretion in response to WI, whereas Pa and Pv increased by 10 +/- 7 and 10.0 +/- 2.0 mmHg relative to the control level, respectively. It is concluded that the reduction of RSNA observed during WI plays a major role in the natriuresis in the dog. PMID- 2563640 TI - Tonic influences from the rostral medulla affect sympathetic nerves differentially. AB - Tonically active neurons in the rostral ventrolateral medulla (RVLM) that project to the autonomic regions of the spinal cord are essential for maintenance of arterial blood pressure at normal levels. Microinjection of glycine into the RVLM in anesthetized cats to inhibit the tonic discharge of these neurons caused variable initial responses in renal and mesenteric nerve discharge and arterial blood pressure. These initial responses were consistently followed by more prolonged decreases in renal and mesenteric nerve discharge and decreases in arterial blood pressure. The tonic influences of neurons in the RVLM were found to be distributed unequally to sympathetic nerves because activity of renal nerves was decreased significantly more than that of mesenteric nerves. The variable nerve and cardiovascular responses during the first 1-3 min after glycine injection were not solely due to loading or unloading of baroreceptors because similar initial responses were seen in vagotomized and sinoaortic denervated cats. Additionally, when muscimol was microinjected into the same sites, only consistent and prolonged decreases in nerve discharge and blood pressure occurred. The inhibitory actions of muscimol on RVLM neurons caused significantly greater decreases in renal than mesenteric nerve activity. Together, these findings demonstrate that the tonic discharge of neurons in the RVLM has unequal influences on renal and mesenteric nerves. PMID- 2563641 TI - Transovarial transmission of Gamboa virus in a tropical mosquito, Aedeomyia squamipennis. AB - We report transovarial transmission of Gamboa virus (Bunyavirus) in Aedeomyia squamipennis, a tropical mosquito which is active and bloodfeeding throughout the year. Gamboa virus was isolated during each of the 28 months of the study from every mosquito stage, including eggs, demonstrating that vertical transmission is a maintenance mechanism of this virus. The overall minimum infection rate was 5.1/1,000 mosquitoes. Identification of the 567 isolates by neutralization indicated that greater than or equal to 2 serotypes or subtypes of Gamboa virus circulate at the study site. PMID- 2563643 TI - Bladder neuropathy in periarteritis nodosa. PMID- 2563644 TI - Pyruvate kinase isozymes from the green alga, Selenastrum minutum. II. Kinetic and regulatory properties. AB - The kinetic and regulatory properties of two pyruvate kinase isozymes, PKp and PKc (apparent chloroplastic and cytosolic isozymes, respectively) from the green alga Selenastrum minutum were studied. The two isozymes differed greatly in several kinetic properties. Although both isozymes showed hyperbolic substrate saturation kinetics, the apparent Michaelis constants for PEP and ADP were about twofold and fourfold lower, respectively, for PKc as compared with PKp. ADP was the preferred nucleotide substrate for both isozymes. However, PKc utilized alternate nucleotides far more effectively than did PKp. PKc and PKp also differed strongly in the effect of activators and inhibitors on the enzymes. Although both isozymes were activated by dihydroxyacetone phosphate (DHAP) with a similar activation constant of about 30 microM, this activator (0.5 mM) caused an approximate 30% increase in the Vmax of PKc, but had no effect on the Vmax of PKp. PKp, but not PKc, was inhibited by ribose 5-phosphate, ribulose 1,5 bisphosphate, 2-phosphoglycerate, phosphoglycolate, and malate. Both isozymes were inhibited by MgATP, Mg2citrate, Mg2oxalate, and Pi. PKc was far more sensitive to inhibition by Pi, as compared with PKp. Pi was a competitive inhibitor of PKc with respect to phosphoenolpyruvate (PEP) (Ki = 1.3 mM). Glutamate was a potent inhibitor of PKc, but had no effect on PKp. In contrast with Pi, glutamate was a mixed-type inhibitor of PKc with respect to PEP (Ki = 0.7 mM). DHAP facilitated the binding of PEP by both isozymes and reversed or relieved the inhibition of PKc by Pi and/or glutamate. The regulatory properties of PKp indicate that it is likely less active in the light and more active in the dark. The in vivo activity of PKc is probably regulated by the relative cytosolic levels of DHAP, Pi, and glutamate; this provides a rationale for the activation of algal cytosolic pyruvate kinase which occurs during periods of enhanced ammonia assimilation. PMID- 2563642 TI - Somatostatin expression in the cerebellar cortex during postnatal development. An immunohistochemical study in the rat. AB - The distribution of somatostatin-immunoreactive (SOM-IR) elements in the cerebellar cortex of the rat has been studied at different stages of postnatal development (from birth to day 30) and in adult animals using immunohistochemistry. The results showed that in vermis of new born animals there are three main groups of SOM-IR structures within the cortex which subsequently spread along the Purkinje cell layer. In addition, both in the vermis and in the lateral lobes, numerous more evenly distributed SOM-positive cells and fibers could be seen. SOM-IR Golgi cells, Purkinje cells and climbing fibers could then be recognized during the subsequent developmental stages. In the vermal zone, SOM IR Purkinje cells formed patches, which seemed to be part of a sagittal columnar or band-like organization. This was most obvious between days 5 and 21 of postnatal development. Subsequently there was a reduction in the number of immunoreactive Purkinje cells but a patchy disposition remained. In addition high numbers of SOM-IR Purkinje and Golgi cells and also climbing fibers were identified in the flocculus and paraflocculus at all stages of development studied, and they were also seen in the adult rats in these regions. In the lateral lobes expression of SOM-like immunoreactivity (LI) decreased and almost completely disappeared in adult animals. The present results demonstrate that a SOM or a SOM-LI peptide can be transiently detected in many Purkinje and Golgi cells in the cerebellar cortex, suggesting a role in events related to developmental processes. However, in some regions and structures SOM-LI can be seen also in adult animals. PMID- 2563646 TI - Prostaglandin E2 and alpha 2 adrenoceptor agonists inhibit the pentose phosphate shunt in pancreatic islets. AB - Glucose utilization in isolated pancreatic islets of the rat was inhibited by prostaglandin (PG) E2 and the alpha 2 adrenoceptor agonist, clonidine, to a similar extent; other prostaglandins did not affect glucose utilization. Islet oxidation of [1-14C]glucose and [6-14C]glucose demonstrated that the pentose phosphate shunt was inhibited by PGE2 and clonidine. Pertussis toxin antagonizes the effects of clonidine and PGE2 on total glucose utilization and pentose phosphate shunt activity. The results suggest that PGE2 and alpha 2 adrenoceptor agonists may regulate glucose metabolism through similar transduction mechanisms, and that a guanine nucleotide binding regulatory (G) protein modulates certain metabolic effects of prostaglandins and adrenergic agonists. PMID- 2563645 TI - Action of Ebselen on rat hepatic microsomal enzyme-catalyzed fatty acid chain elongation, desaturation, and drug biotransformation. AB - In the previous study, the organoselenium-containing anti-inflammatory agent, Ebselen, was found to disrupt both hepatic microsomal NADH- and NADPH-dependent electron transport chains. In the current investigation, we focus on the action of Ebselen on three separate metabolic reactions, namely, fatty acid chain elongation, desaturation, and drug biotransformation, which utilize reducing equivalents via these microsomal electron transport pathways. Both NADH-dependent and NADPH-dependent chain elongation reactions showed (i) that the condensation step was inhibited by Ebselen; all three substrates, palmitoyl CoA (16:0), palmitoleoyl CoA (16:1), and gamma-linolenyl CoA (18:3), were differentially affected by Ebselen; for example, the apparent Ki's of Ebselen for the condensation of 16:0, 16:1, and 18:3 in the absence of bovine serum albumin (BSA) preincubation were 7, 14, and 34 microM, and those in the presence of BSA preincubation were 35, 62, and 150 microM, respectively, supporting earlier data for multiple condensing enzymes; (ii) that the beta-ketoacyl CoA reductase catalyzed reaction step which appears to receive electrons, at least in part, from the cytochrome b5 system, was also markedly inhibited by varying Ebselen concentrations; and (iii) that similar results were obtained with the dehydrase and the enoyl CoA reductase. Hence, each of the four component steps was significantly inhibited by Ebselen. Another important fatty acid biotransformation reaction, delta 9 desaturation of stearoyl CoA to oleoyl CoA, was significantly inhibited (90%) by 30 microM Ebselen. This effect appeared to be directly related to the NADH-dependent electron transport chain rather than to a direct action on the desaturase enzyme. Last, Ebselen also inhibited both aminopyrine and benzphetamine N-demethylations, two cytochrome P450-catalyzed reactions, in untreated rats, in rats on a high carbohydrate diet, and in phenobarbital-treated rats. PMID- 2563647 TI - Elevated blood pressure in treated hypertensives with low-level lead accumulation. AB - The relationship between blood pressure (BP) and blood lead concentration (PbB) was examined in 51 bus drivers who were treated for hypertension. These drivers were a subset of a representative sample (N = 342) of the driver population (N = approximately 2,000), and were not selected for hypertension or lead exposure. Blood lead concentrations ranged from 2-24 micrograms/dl (median: 6.9 micrograms/ld). There were 33 subjects treated primarily with diuretics, and 18 subjects were treated with beta blockers. Adjusted regression coefficients relating systolic BP with PbB were -6.4 +/- 11.4 and 4.5 +/- 12.9 mmHg/In(micrograms/dl) in each group, respectively, but were not statistically significant. The adjusted coefficients for diastolic BP were 1.12 +/- 3.89 and 14.3 +/- 5.69 mmHg/In(micrograms/dl) (p = 0.036), respectively. The latter relationship represents an average increment of 12 mmHg in diastolic BP over the range of observed PbBs (2.0 to 11.4 micrograms/dl) in subjects treated with beta blockers. Thus, beta blocker therapy may be less effective in reducing diastolic pressure in individuals with higher PbBs and suggests an action of lead at PbBs below current standards. PMID- 2563648 TI - Mental testing with metoprolol. PMID- 2563649 TI - Beta-blockade and mental performance. PMID- 2563650 TI - Mechanism of activation of particulate guanylate cyclase by atrial natriuretic peptide as deduced from radiation inactivation analysis. AB - The interaction between the receptor (Rc) for atrial natriuretic peptide (ANP) and the effector enzyme particulate guanylate cyclase (GC) has been studied by radiation inactivation. Irradiation of bovine lung membranes produced an increase in GC activity at low radiation doses followed by a dose-dependent reduction at higher doses. This deviation from linearity in the inactivation curve disappeared when lung membranes were pretreated with ANP. Essentially identical results were also obtained with adrenal membranes. Based on these radiation inactivation data, the following dissociative mechanism of activation of particulate guanylate cyclase by ANP has been proposed: Rc.GC(inactive) + ANP----Rc.ANP + GC(active). PMID- 2563651 TI - Inhibition of purified human postheparin lipoprotein lipase by beta-adrenergic blockers in vitro. AB - We examined the effects of five beta-adrenergic blockers on the hydrolysis of phosphatidylcholine-stabilized triolein particles by purified human postheparin lipoprotein lipase (PHLpL) in order to evaluate the possible role of direct inhibition as a mechanism of drug-induced hypertriglyceridemia. The relative inhibitory potencies were observed in the following order: propranolol much greater than pindolol greater than metoprolol greater than atenolol greater than nadolol. There was a positive correlation between the octanol/water partition coefficients of these agents and their inhibition of lipoprotein lipase, suggesting that hydrophobicity may be one of the major determinants for PHLpL inhibition. The amount of the beta-adrenergic blockers required to produce 50% inhibition of human PHLpL was much greater than that required to inhibit purified bovine lipoprotein lipase. PMID- 2563652 TI - High affinity binding of 3H rauwolscine and 3H RX781094 to alpha 2 adrenergic receptors and non-stereoselective sites in human and rabbit brain cortex membranes. AB - The radiolabeled antagonists 3H RX 781094 and 3H rauwolscine bind with high affinity to alpha 2 adrenergic receptors as well as to non-receptor sites in human and rabbit brain cortex membranes. These non-receptor sites form an important contaminant of the specific binding when non-specific binding is determined in the presence of 10 microM phentolamine or more. While phentolamine is no suitable ligand to discriminate both sites, (-)-epinephrine displays a sufficient affinity ratio to separate radioligand binding to these sites. When 1 microM (-)-epinephrine is used for the determination of the non-specific binding, both radioligands bind specifically to alpha 2 receptors. Under these conditions, 3H rauwolscine and 3H RX 781094 bind to the same amount of non-cooperative sites; binding isotherms for human brain are Bmax = 113 +/- 15 fmol/mg protein and Kd = 22.8 +/- 4.2 nM for 3H RX781094 and Bmax = 110 +/- 17 fmol/mg protein and Kd = 4.7 +/- 2.5 nM for 3H rauwolscine. Competition binding experiments show, for both radioligands and in both species, the typical pharmacological potency order of alpha 2 adrenergic receptors, i.e. phentolamine greater than yohimbine greater than prazosin for the antagonists and UK 14304 greater than p-aminoclonidine greater than or equal to (-)-epinephrine greater than (+)-epinephrine greater than isoproterenol for the agonists. Whereas the alpha 2 receptor sites display high affinity and stereoselectivity towards (-)-epinephrine and (+)-epinephrine, the non-receptor sites bind both epinephrine isomers with equal low affinity. Specific binding of both radioligands to these sites can be determined when total binding is performed in the presence of 1 microM (-)-epinephrine and non-specific binding the presence of 1 mM phentolamine. 3H rauwolscine binding to the non stereoselective sites can be displaced with high affinity by 5-HT, suggesting binding to a 5-HT1-receptor. The 3H RX 781094 binding displays low affinity for most alpha adrenergic ligands and do not correspond to beta adrenergic, dopaminergic or serotonergic receptors. PMID- 2563653 TI - Pharmacology of binding of 3H-SCH-23390 to D-1 dopaminergic receptor sites in rat striatal tissue. AB - 3H-SCH-23390, a selective antagonist of D-1 dopamine (DA) receptors, was used in a radioreceptor assay with rat brain striatal tissue, optimized biochemically, and extensively characterized pharmacologically with striatal membranes. Nonspecific binding, defined with excess cis(Z)-flupenthixol (300 nM), averaged 20-25% of total counts bound. Specific binding was linearly dependent on the amount of original striatal tissue (0-4 mg) or protein (0-250 micrograms), temperature dependent, saturable and reversible, and appeared to involve a single site at ligand concentrations limited to less than 10 nM. Binding in rat brain regions ranked as: striatum greater than accumbens greater than prefrontal cortex greater than posterior cerebral cortex greater than cerebellum. Association was virtually complete within 30 min at 30 degrees, and the rate of dissociation at 30 degrees was 0.0377 min-1 (half-time = 18.4 min). Affinity (Ka or Kd) determined from association and dissociation rate constants and from concentration isotherms averaged 0.349 and 0.340 nM respectively. Including Na+ at 150 mM increased apparent maximum specific binding (Bmax) by 19%, with a 29% increase in affinity; other monovalent cations alone had small effects on specific binding; Ca2+ and Mg2+ reduced binding by 42%. Agents (N = 85) were tested for potency (Ki or IC50) in competition with the ligand (at 0.30 nM). Those known to have selective effects at D-1 receptors, generally, were most potent and stereoselective. Na+ (150 mM) had little effect on the affinity of cis thioxanthenes but decreased that of most other agents tested with high D-1 affinity. For antipsychotic agents, the correlation of typical clinical daily doses versus Ki at D-1 sites (r = 0.06) was much lower than at D-2 sites (r = 0.94). (-)Thioridazine was discovered to be D-1 selective, whereas the (+) enantiomer was selective for D-2 sites labeled with 3H-spiperone. Relatively sedating antidepressants had greater D-1 affinity than their less-sedating, secondary amine congeners. PMID- 2563654 TI - Multiple mechanisms of adriamycin resistance in the human leukemia cell line CCRF CEM. AB - CEM cells exhibiting a 25-fold (C25X) or 80-fold (C80X) increase in resistance to adriamycin were isolated and characterized. C25X cells were cross-resistant to daunomycin and etoposide (VP-16) but not to vincristine or colchicine. These cells were not defective in the cellular accumulation of drug and did not contain detectable levels of P-glycoprotein. Continued exposure of C25X cells to adriamycin resulted in increased levels of resistance and additional phenotypic changes. These cells (C80X) now contained high levels of P-glycoprotein and were cross-resistant to a variety of agents including vincristine and colchicine. A fluorometric assay for DNA unwinding was used to measure levels of drug-induced DNA breaks in sensitive and C25X resistant cells. Studies carried out with VP-16, 4'9-acridinyl-aminomethanesulfon-m-anisidide (m-AMSA), adriamycin, or daunomycin showed that the level of drug-induced DNA strand breakage in resistant cells was considerably less than that occurring in drug-treated sensitive cells. These studies, therefore, show that treatment of CEM cells with adriamycin resulted in a nuclear alteration that contributed to drug resistance. They also demonstrate that prolonged treatment of cells with adriamycin resulted in membrane alterations that affect cellular drug accumulation. Adriamycin resistance in CEM cells can thus occur as a result of at least two distinct mechanisms. PMID- 2563655 TI - Analysis of structural features of dihydropyridine analogs needed to reverse multidrug resistance and to inhibit photoaffinity labeling of P-glycoprotein. AB - Synthetic dihydropyridine analogs were screened to determine whether they would reverse multidrug resistance of a multidrug-resistant human KB carcinoma cell line, KB-C1. Among twenty-four dihydropyridine analogs examined, thirteen almost completely overcame drug resistance (group A), nine partially overcame resistance (group B) and two did not reverse resistance (group C). The twenty-two compounds that reversed drug-resistance (groups A and B) were hydrophobic dihydropyridine derivatives. Three compounds that reversed resistance, NK-113, NK-138 and NK-194, increased the accumulation of [3H]vincristine in the resistant KB-C1 cells, but not in the parental KB cells, nor in a revertant cell line, KB-C1-R2. NK-101 (group C), which did not reverse resistance, had no effect on drug accumulation. Enhanced efflux of vincristine from the resistant cells was inhibited completely by NK-194, but NK-194 did not affect vincristine influx. Nine of the twenty-four compounds were screened to determine whether they inhibited photoaffinity labeling of the cell surface protein gp170 (P-glycoprotein) in KB-C1 cells by N (p-azido-[3-125I]-salicyl)-N'-beta-aminoethylvindesine [( 125I]NASV). All five compounds of group A, NK-138, NK-194, NK-200, NK-203 and NK-220, inhibited the photoaffinity labeling of gp170 at less than 10-100 microM, whereas NK-113 and NK 196 of group B inhibited the labeling at 100-200 microM. By contrast, NK-101 and NK-102 of group C did not inhibit labeling even at 2000 microM. These studies confirm the relationship among reversal of multidrug resistance, decreased efflux of vincristine, and inhibition of [125I]NASV labeling of P-glycoprotein. PMID- 2563656 TI - Glucocorticoid hormones prevent the induction of gamma-glutamyl transpeptidase by ethanol in a rat hepatoma cell line. AB - The increase in serum gamma-glutamyl transpeptidase (GGT) is a well known marker of chronic alcoholism in man. We have previously shown that ethanol (180 mM) induces GGT activity 2-3-fold in the C2 rat hepatoma cell line. In this study, we have analyzed the interaction of ethanol with steroid hormones and drugs in this well defined cell culture system. Dexamethasone (100 nM), a synthetic glucocorticoid agonist, completely prevented the induction of GGT by ethanol, but had no effect when added alone. This inhibitory effect was also observed with other corticosteroids, but not with sex steroids; it was prevented by RU 486, a glucocorticoid antagonist. These observations suggest that dexamethasone acts through a high affinity glucocorticoid receptor. Conversely, ethanol did not interfere with the glucocorticoid induction of alanine aminotransferase in the same cell. We have analyzed the metabolism of ethanol in the C2 cells. These cells lack significant alcohol dehydrogenase activity as well as any cytochrome P 450 Alc immunoreactivity. Dexamethasone did not modify the disappearance of ethanol in the culture medium of those cells. We conclude that glucocorticoid hormones interact with ethanol at the cellular level, and that this interaction does not involve a modification of alcohol metabolism. PMID- 2563657 TI - Treatment of pain in acute myocardial infarction. AB - The treatment of pain in acute myocardial infarction varies with local practice. Narcotic analgesics are still the usual treatment in many hospitals. Knowledge of optimal doses, duration of pain relief, and time between drug administration and pain relief is inadequate. Many studies indicate that the relief of pain is often incomplete after treatment with narcotic analgesics. There is often a need for alternative treatments. Large randomised studies consistently show that beta blockade, initially given intravenously and then orally, relieves pain and reduces the need for analgesics. Studies also indicate that early administration of streptokinase and glyceryl trinitrate relieves pain. There is evidence that drugs that limit ischaemic damage also relieve pain. PMID- 2563658 TI - Median nerve evoked potentials during propofol anaesthesia. AB - Median nerve somatosensory evoked potentials (SSEP) were studied in 10 ASA I patients during general anaesthesia with propofol, vecuronium and fentanyl after premedication with diazepam. N14 and N20 latencies, central conduction time (CCT) and corresponding interpeak amplitudes were assessed at induction, after intubation, during steady state, at re-commencement of spontaneous breathing and during early recovery. Data were compared with pre-induction values. Median nerve SSEP were reproducible throughout the procedure and wave forms were suitable for assessment of neurological disorders. Intra-individual changes were considerable for latencies and amplitudes. PMID- 2563659 TI - Circulating stem cell autografts in high-risk myeloma. PMID- 2563660 TI - Metabolism of long-chain polyunsaturated fatty acids in isolated cardiac myocytes. AB - The uptake and integrated intracellular metabolism of (n - 6) and (n - 3) polyunsaturated fatty acids was studied in isolated rat cardiac myocytes and in the perfused heart. Labeled linolenic acid (18:3(n - 3)) uptake and its subsequent metabolism into carbon dioxide as well as acylation into lipids was nonsaturable over a substrate range of 0.02 to 0.4 mM. [1-14C]Linoleic acid (18:2(n - 6)), dihomo-gamma-linolenic acid (20:3(n - 6)) and arachidonic acid (20:4(n - 6)) were transported into myocytes at rates similar to those for linolenic acid. Conversely both [1-14C]-gamma-linolenic acid (18:3(n - 6)) and eicosapentaenoic acid (20:5(n - 3)) were taken up at a slower rate. Oxidation of 18:3(n - 6) was 4-5-fold greater when compared with C18-C20 polyunsaturated fatty acids. When myocytes were incubated with labeled 18:2(n - 6), 18:3(n - 6), 18:3(n - 3), 20:4(n - 6) or 20:5(n - 3), it was not possible to detect any desaturation or chain-elongation products. Identical results were obtained when hearts were perfused with 1-14C-labeled linoleic acid. PMID- 2563661 TI - Two phases of engraftment established by serial bone marrow transplantation in mice. AB - Serially transplanted bone marrow eventually fails to reconstitute lethally irradiated mice. The reasons for this loss of repopulating ability are unknown. We showed that serial bone marrow transplantation changed the ratio of hematopoietic progenitors in bone marrow. The numbers of granulocyte-macrophage colony-forming units (CFU-GM) in the bone marrow did not change with serial transplantation. Spleen CFU (CFU-S) numbers decreased with serial transfer but remained at levels which should be associated with engraftment, even on the transfers which were unsuccessful. The CFU-S, therefore, did not appear to be the cells responsible for long-term hematopoietic repopulation. The number of successful serial transfers was dependent on the size of the grafts, and prolonging the time interval between transfers reestablished the ability of the serially transplanted marrow to reconstitute lethally irradiated recipients. Serial bone marrow transplantation dissociated two phases of engraftment. The first unsustained phase was maintained with repeated serial transfer and appeared to be produced by committed progenitors. The second sustained phase was eventually lost with repeated serial transfer and was apparently due to the pluripotent stem cell. PMID- 2563662 TI - Effect of 5-fluorouracil on "primitive" hematopoietic stem cells that reconstitute whole erythropoiesis of genetically anemic W/Wv mice. AB - The potential to reconstitute the whole erythropoiesis of a genetically anemic (WB X C57BL/6)F1-W/Wv (WBB6F1-W/Wv) mouse for at least 8 weeks was compared between 5-fluorouracil (5FU)-treated and nontreated bone marrow cells. C57BL/6 Pgk-1b/Pgk-1a female mice, in which each stem cell had either A-type or B-type phosphoglycerate kinase (PGK) owing to the random inactivation of one of two X chromosomes, were used as donors. As a marker of the reconstitution, electrophoretic pattern of hemoglobin was used. The concentration of the stem cells that reconstitute the whole erythropoiesis of WBB6F1-W/Wv mouse was higher in the marrow of donors that had received an injection of 5FU two days previously (two-day 5FU-treated) than in the marrow of nontreated donors. In the marrow of four-day 5FU-treated mice, however, the concentration was comparable to that of nontreated mice. The PGK electrophoretic pattern of WBB6F1-W/Wv mice reconstituted by nontreated marrow cells was comparable to the PGK pattern of WBB6F1-W/Wv mice reconstituted by four-day 5FU-treated marrow cells. Thus, a single stem cell with extensive proliferative potential rather than multiple spleen colony-forming units appeared to be responsible for the erythropoietic reconstitution in the transplantation of nontreated healthy marrow cells as well as 5FU-treated marrow cells. PMID- 2563663 TI - Molecular defect in factor IXHilo, a hemophilia Bm variant: Arg----Gln at the carboxyterminal cleavage site of the activation peptide. AB - A genomic DNA library and the enzymatic DNA amplification technique were used to isolate human factor IX coding sequences of a hemophilia Bm variant, factor IXHilo. A point mutation that resulted in the substitution of a glutamine (CAG) for an arginine (CGG) at amino acid 180 was found in exon VI of the factor IX gene (G----A at nucleotide 20519). This mutation alters the carboxy terminal cleavage site for the activation peptide at Arg180-Val181. The arginine residue at the activation peptide cleavage site is conserved in mouse, canine, bovine, and human factor IX, suggesting that the arginine at amino acid 180 is important for normal cleavage. Sequencing of all of the coding regions of factor IXHilo revealed no other mutations. We have also shown that the point mutation in exon VI creates a new Dde I restriction site, which, in combination with the enzymatic DNA amplification technique, provides a quick, reliable, and sensitive method for carrier detection and antenatal diagnosis in affected kindreds. This is the first report of the molecular defect in a hemophilia Bm patient with a markedly prolonged ox brain prothrombin time. PMID- 2563665 TI - A PvuII polymorphism of the bcr region in patients with hematopoietic disorders and their families. AB - The BCR gene on chromosome 22 has received increasing attention because of its involvement in the Philadelphia (Ph') translocation. For most restriction enzymes, this locus has been found to be nonpolymorphic. Two alleles have only been found when Taql-digested DNA is hybridized to a 5' bcr-specific probe. We describe another two-allele polymorphism detected by the same probe in PvuII digested DNA. The polymorphism is characterized by an additional PvuII site in the bcr region: this causes the appearance of an additional band of about 2.3 kb or 2.5 kb besides a 4.8-kb fragment in hybridizations with the 5' bcr or a 3' bcr probe. The incidence of the second allele is very low. It has only been found in some patients with hematopoietic malignancies and in a group of volunteers having a leukemia patient in their families. PMID- 2563666 TI - Paul and Carol--1. An introduction to modern molecular genetics. PMID- 2563664 TI - Immunohistochemical detection and quantitation of P-glycoprotein in multiple drug resistant human myeloma cells: association with level of drug resistance and drug accumulation. AB - Using several multiple drug-resistant human myeloma cell lines as standards, we developed an immunohistochemical staining technique and means of quantitating P glycoprotein in individual myeloma cells. The level of staining intensity for P glycoprotein in individual myeloma cells was quantitated by measuring the average optical density of each cell with a microscopic computerized cell analysis system. Using this system, we observed that the level of P-glycoprotein for individual cells within a cell population of known drug sensitivity was very homogeneous (coefficient of variation less than or equal to 13%). Analysis of cell lines with gradually increasing levels of multidrug resistance (8226/S, 8226/Dox6 and 8226/Dox40) demonstrated a close association between the level of resistance to doxorubicin, defined by the mean lethal dose (D0) and the amount of P-glycoprotein on individual cells determined by the optical density (r = 0.82, P less than 0.0005). Intracellular doxorubicin (DOX) accumulation in the individual cell lines was inversely related to the level of drug resistance expressed as D0. P-glycoprotein was also detected in the marrow-derived myeloma cells of patients with drug refractory disease using immunohistochemical staining. The amount of P glycoprotein in the cells of one patient was directly compared to the amount found in the simultaneously stained standard cell lines (8226/Dox6 and 8226/Dox40) by comparing the optical densities for individual cells. Using this immunohistochemical technique to detect and quantitate P-glycoprotein in patient myeloma cells and comparing it to standard multidrug resistant myeloma cell lines may be of value in determining the contribution of P-glycoprotein to clinical drug resistance in patients with multiple myeloma. PMID- 2563669 TI - Clinical characteristics in sporadic and familial medullary thyroid carcinoma. A nationwide study of 249 patients in Sweden from 1959 through 1981. AB - All patients with medullary thyroid carcinoma (MTC) diagnosed in Sweden during 1959 through 1981 were recruited for study from the Swedish Cancer Registry. Among a total of 249 patients, 66 were diagnosed in 1959 through 1969 and 183 from 1970 through 1981. Apparently sporadic MTC was present in 186 patients, and familial MTC in 63. Twenty-seven patients with familial MTC were diagnosed from clinical symptoms and 36 by screening. In both the sporadic group and the symptomatic familial group, approximately 80% of the patients had palpable thyroid tumors. Lymph node metastases were present in 44% of the sporadic group and in 37% of the group with symptomatic familial MTC, and distant metastases in approximately 20% of the patients in each of these groups. The patients detected by screening differed significantly from the two groups of symptomatic cases by having a lower frequency of palpable thyroid lesions (50%), smaller tumors, and lower frequencies of cervical lymph node metastases (14%) and distant metastases (0%). Multivariate analyses revealed no significant differences between sporadic cases and symptomatic familial cases regarding tumor size or stage. Large tumors (greater than 3 cm) were more frequently accompanied by palpable cervical lymph nodes and were associated with an approximately four times higher risk of distant metastases than tumors smaller than 1 cm. Women had significantly smaller tumors and a more favorable stage distribution than age-related men. PMID- 2563667 TI - Parkinson's disease. AB - In Parkinson's disease there is degeneration of neurons in the substantia nigra, with consequent depletion of the neurotransmitter dopamine. The triad of tremor, rigidity and bradykinesia is the clinical hallmark. Drugs currently used for palliative therapy fall into three categories: anticholinergic agents, dopamine precursors (levodopa combined with extracerebral decarboxylase inhibitors) and artificial dopamine agonists. It has been argued, on theoretical grounds, that some drugs slow the progress of Parkinson's disease, although no firm evidence has supported this. Treatment must be individualized, and more than one type of drug can be given concurrently after a careful build-up in dosage. We review the adverse effects of various drugs and consider new developments such as slow release preparations, selective D-1 and D-2 agonists and transplants of dopaminergic cells into the brain. The treatment of Parkinson's disease can be demanding, rewarding and sometimes frustrating, but it remains a most challenging exercise in pharmacotherapy. PMID- 2563668 TI - Somatostatin in medullary thyroid cancer. In vitro and in vivo studies. AB - The authors evaluated the presence of somatostatin (SRIF) in the plasma and in the tumor tissue of a total of 22 patients with medullary thyroid cancer (MTC) and studied the effect of exogenous SRIF administration on basal and pentagastrin (PG)-stimulated plasma calcitonin (CT) and carcinoembryonic antigen (CEA) levels. Mean plasma SRIF concentrations were significantly higher than those found in normal controls, with five of 15 patients having plasma SRIF levels above the mean + 2 SD of normal controls. High immunoreactive SRIF concentrations were found in the extract of three tumor tissues but not in one follicular thyroid cancer or in one toxic diffuse goiter. By immunoperoxidase staining seven of 11 (63.6%) primary MTC and five of 13 (38.5%) metastases expressed SRIF antigen in a low number of cells and with a weak degree of staining. As expected, CT was expressed in almost 100% of the cases with positivity in most of the cells and strong degree of staining. Patients with positive SRIF staining in the primary tumor had longer survival than SRIF negative patients. Infusion of synthetic SRIF (11 micrograms/minute/45 minutes) produced a significant reduction of plasma CT (but not CEA) levels in 12 of the 15 patients submitted to this test. Maximal percent decrease of plasma CT ranged from 10.8% to 72.7% of the basal value and was usually observed between 30 and 45 minutes from the beginning of the infusion. When infused together with the injection of PG, SRIF was able to significantly (P less than 0.05) inhibit the PG-induced CT release in five of six patients tested. These results demonstrate the following: SRIF is present in a few cells of many primary MTC and less frequently in their metastases; tentatively, the expression of SRIF antigen in the tumor seems to be associated with longer survival; increased SRIF concentrations are found in the plasma of some patients with metastatic involvement; and treatment with exogenous SRIF reduces the basal and PG-induced CT (but not CEA) release from the tumor. PMID- 2563670 TI - Ring chromosome in a patient with MEN IIA S. AB - Three patients with MEN IIA S belonging to the same family, which has five affected subjects in two generations, have been studied. Constitutional karyotype examination identified an extra ring chromosome in 3% of the cells in one of the affected subjects. The number of rings for each cell varied, and different evolutionary aspects of the ring, such as doubling and interlocking, were present. Furthermore, asynchronous DNA replication of doubling rings was observed. This nonhomogeneous, nonrandom chromosome abnormality found in normal tissue of patients with MEN IIA S could be indicative of sensitivity to structural alteration of some chromosome regions. PMID- 2563671 TI - Frequent loss of heterozygosity on chromosome 14q in neuroblastoma. AB - Using 29 polymorphic DNA markers which detect allelic deletion of genes at specific loci on 19 different chromosomes, we analyzed 14 neuroblastomas for possible loss of chromosomal heterozygosity. The incidence of loss of heterozygosity was high at the D14S1 locus on chromosome 14q, being detected in six of 12 patients (50%). In spite of the cytogenetic finding suggesting high frequency of chromosome 1p deletion, loss of heterozygosity at the MYCL locus on 1p32 was detected only in two of nine patients (22%). It was also found in two of 11 patients (18%) on 13q, but not on chromosomes 2, 3, 5, 6, 7, 8, 9, 10, 11, 12, 15, 16, 17, 18, 19, and 20. The present results indicate that recessive genetic changes involving sequences on chromosome 14q may play an important role in the development of neuroblastoma. PMID- 2563672 TI - Phenotypic instability of drug sensitivity in a human colon carcinoma cell line. AB - Colon cancer is one of the tumors most refractory to treatment by chemotherapy, and this may be due to inherent phenotypic instability of such tumor cells with respect to the biochemical determinants of drug sensitivity. To test this hypothesis, a clonal human colon carcinoma cell line, clone A, was passaged in culture in the absence of selection conditions or mutagens. During this time, sensitivity to several drugs was examined, and was found to decrease 4-fold during 30 weeks of culture. Five randomly selected subclones, having never been exposed to drug or mutagen, displayed a range of sensitivities to etoposide (50% inhibitory concentrations ranging from 1.5 to 4.9 microM) and to vincristine (9 fold range), but all had the same sensitivity to methotrexate. With time these sensitivities also changed, and subsequent subclones were chosen from the lines with highest and lowest drug sensitivity. Again a wide range of phenotypes was observed. Sensitivity to vincristine ranged 14-fold and to doxorubicin 3-fold. Several biochemical determinants of drug sensitivity had a broad range of expression between cell lines. Cellular accumulation of [3H]vincristine, as well as expression of multidrug resistance protein P170 and glutathione transferase activity all varied significantly between subclonal lines. This suggests that some human colon tumors may be phenotypically unstable with respect to drug sensitivity, and this could contribute to clinical resistance to chemotherapeutic compounds. PMID- 2563673 TI - Synergistic activation and repression of transcription by Drosophila homeobox proteins. AB - We have used a transient expression assay employing Drosophila tissue culture cells to study the potential of several Drosophila homeobox proteins to function as transcriptional regulators. A 96 bp fragment from the promoter region of the segment polarity gene engrailed, previously shown to contain five copies of a 10 bp consensus binding site for these proteins, enhanced transcription in the presence, but not the absence, of several different homeobox protein expression vectors. It is interesting that cotransfection with combinations of expression vectors encoding the homeobox proteins fushi tarazu, paired, and/or zen resulted in substantial synergistic increases in expression. In contrast, the products of the even-skipped and engrailed genes were found to repress, or quench, the activation induced by the other proteins. We discuss the implications of these results with respect to the role of homeobox genes in the control of embryonic development, and propose a "multi-switch" model whereby the activity of a target gene depends on the interactions of different homeobox proteins with multiple copies of a common binding site. PMID- 2563674 TI - Pressor hormones regulate atrial-stretch-induced release of atrial natriuretic peptide in the pithed rat. AB - Atrial wall stretching is a known stimulus for atrial natriuretic peptide (ANP) secretion. The effects of the stimulation of autonomic nervous system, hemodynamic factors, and humoral factors (epinephrine, angiotensin, vasopressin, and brain extracts) on the release of ANP under basal conditions and during increased atrial pressure produced by acute volume loading in pithed rats were examined. In conscious rats, acute volume expansion by 0.9% of saline (4 ml) increased the plasma immunoreactive ANP (IR-ANP) concentrations by a factor of 4 (140 +/- 30 pg/ml vs. 521 +/- 140 pg/ml, p less than 0.001, n = 8), whereas volume-induced ANP release was blocked in pithed rats (75 +/- 9 pg/ml vs. 99 +/- 13 pg/ml, NS, n = 7). The ANP versus right atrial pressure curve shifted to the right, indicating that much smaller amounts of IR-ANP were released in pithed than in conscious rats for each given increase in right atrial pressure. Electrical vagal and sympathetic nerve stimulation or changes in heart rate had no effect on plasma IR-ANP concentrations and failed to restore the volume-load induced release of ANP in pithed rats. When extracts of anterior pituitary lobe, brain cortex, or hypothalamus were infused, no effect on volume-expansion-induced plasma IR-ANP levels was seen. In contrast, acute volume expansion caused a fourfold increase in levels of circulating IR-ANP in pithed rats that received posterior pituitary extracts, and the ANP versus right atrial pressure curve shifted markedly to the left. Infusion of a V1 antagonist blocked the volume expansion-induced ANP release produced by the posterior pituitary extract. When [Arg8]-vasopressin (0.025 or 0.05 micrograms/kg/min) was infused to pithed rats, mean arterial pressure increased but basal plasma IR-ANP did not change significantly. However, acute volume expansion in the presence of vasopressin infusion (0.05 micrograms/kg/min) increased the amount of circulating IR-ANP by a factor of 4 (113 +/- 14 pg/ml vs. 414 +/- 43 pg/ml, p less than 0.001, n = 8). Thus, for a given increase in right atrial pressure, a similar amount of IR-ANP was released in the pithed rat during the vasopressin infusion as in the normal conscious animal. V1 antagonist blocked the increase in mean aterial pressure as well as the increase of plasma IR-ANP produced by [Arg8]-vasopressin. In addition, volume expansion during intravenous epinephrine (1.75 micrograms/kg/min) and angiotensin (1.0 micrograms/kg/min) doubled plasma IR-ANP levels.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2563676 TI - Ontogeny of hypothalamic vasopressin, oxytocin and somatostatin gene expression. AB - The expression of 3 neuropeptide genes, vasopressin (AVP), oxytocin (OT) and somatostatin (SOM), was studied in the developing rat hypothalamus using Northern blot analysis combined with densitometric scanning. A unique profile of developmental expression was established for each of the 3 genes. SOM mRNA is detectable at embryonic day 14 and reaches 40% of the adult levels by embryonic day 18. By contrast, accumulation of AVP and OT mRNA is mainly a postnatal event. AVP mRNA, although detectable in the late embryo, rises gradually after birth and attains 40% of adult levels after the second postnatal week. Maturation of OT gene expression occurs even later and parallels AVP gene expression with a lag time of one week. Observed increases in mRNA levels are due to an upregulation of gene expression since they occur essentially following cessation of neuronal cell proliferation. The rise in AVP and OT mRNA accumulation coincides with the establishment of synaptic input to AVP and OT neurons. Expression of the SOM gene, by contrast, occurs prior to neuronal cell differentiation and points to a possible function of SOM in the embryonic brain. PMID- 2563677 TI - Molecular genetics and craniofacial malformations. PMID- 2563675 TI - Ontogeny of somatostatin in cerebral cortex of macaque monkey: an immunohistochemical study. AB - Distribution of somatostatin-immunoreactive cells in the cerebral cortex of macaque monkeys at embryonic day 120 (E120), E140, newborn, postnatal day 60 (P60) and adult stages were studied by the avidin-biotin-peroxidase immunohistochemical method. At all stages, there existed 3 types of cells in the gray matter: bipolar, multipolar and small-sized cells which stained only in perikaryon. Somatostatin-immunoreactive cells were observed from E120. The cell number increased between E120 and E140 and decreased until P60. At the newborn stage, a high density of cells was distributed in layer II of the prefrontal and parietal cortices (areas FD and PE). In layer I of the postcentral, parietal, temporal and preoccipital cortices (areas FA, PC, PE, TA, TE and OA), small numbers of horizontal cells were detected only at the embryonic and newborn stages. In adulthood, the number of somatostatin cells was much smaller than at the early stages (E140 and newborn). Compared to other cortical areas, in occipital cortex (area OC), there was little change in cell number during development. In occipito-temporal cortices, there were increases in cell number from posterior to anterior portion at all the stages. The large number of somatostatin cells in all layers of the cerebral cortex during the early stages indicates that somatostatin plays a role in the development of the monkey cerebral cortex. PMID- 2563678 TI - X-linked cleft palate and ankyloglossia in an Icelandic family. AB - Information was available on 293 family members and spouses of seven generations in an Icelandic family with high frequency of cleft of secondary palate and ankyloglossia. The authors have personally investigated 182 individuals in generations IV-VII and have drawn blood from over 100 members for genetic marker studies. The senior author, Dr. Bjornsson, has operated on two-thirds of the affected individuals. Twenty-six family members had cleft palate (CP) and, of these, 19 (17 male and two females) had ankyloglossia as well (CP + A). Twenty females and one male had only ankyloglossia (A). All mothers in one of two branches of the family who had sons with CP + A had ankyloglossia themselves. This was not the case in the other branch, in which the mothers of affected sons were themselves unaffected. Fathers affected with CP, CP + A, or high vaulted palate (HVP) never had affected sons. As reported earlier, the condition has been mapped to the q13-q21 region of the X chromosome using restriction fragment length polymorphism (RFLP) techniques (Moore et al, 1987). Our conclusion is that this midline defect is X-linked but varies in the severity of expression. PMID- 2563679 TI - New techniques in the management of foot trauma. AB - Modern techniques are effective in the reduction and fixation of intraarticular fractures of the foot. While not all fractures can be restored anatomically, careful evaluation of roentgenograms and computed tomographic scans will identify those for which open reduction and internal fixation may be considered. Careful attention to the biomechanical principles and surgical techniques of open reduction and internal fixation can restore function and a useful, painless foot in previously hopeless cases. PMID- 2563680 TI - Common dental emergencies. Evaluation and management for emergency physicians. AB - Dental pain and disfigurements cause patients to seek care from the Emergency Department of hospitals. It is important for physicians to understand dental disease and trauma in order to diagnose, treat, and refer patients with dental emergencies efficiently. Many common dental problems have been discussed in clinical terms. Treatment and referral options were offered. Techniques for local dental anesthesia and avulsed tooth splinting were described. PMID- 2563681 TI - Hormone ontogeny in the ovine fetus and neonate. XXII. The effect of somatostatin on the growth hormone (GH) response to GH-releasing factor. AB - We postulated that an increase in the biological effectiveness of somatostatin (SRIF) accounts, at least in part, for the decrease in basal and GRF-induced ovine GH (oGH) secretion observed around birth in the ovine fetus and neonate. To test this hypothesis, SRIF (SRIF-14; given as 30 micrograms/kg iv bolus, followed by 2 micrograms/kg.min for 75 min) was infused into chronically catheterized fetal and neonatal lambs, and the oGH response induced by GRF [GRF-(1-44) amide; 1 microgram/kg] in the presence of exogenous SRIF was compared to the oGH response induced by GRF in saline-infused controls. In fetuses of 115-122 days gestation, SRIF had no detectable effect on the oGH response to GRF [peak incremental oGH response (mean +/- SEM), 527 +/- 124 vs. 562 +/- 103 ng/ml in controls]. In neonatal lambs (3-17 days old), SRIF completely suppressed the immediate oGH response to GRF (peak incremental response, 0.8 +/- 1.3 vs. 111 +/- 34 ng/ml in controls; P less than 0.02). In late gestational fetuses (126-139 days old), a transitional pattern was observed (peak incremental oGH response, 207 +/- 56 vs. 324 +/- 30 ng/ml in controls; P less than 0.04). In the second part of this study, we explored, in the neonatal lamb, the hypothesis that SRIF withdrawal plays a role in pulsatile GH secretion and that the amount of GRF to which the somatotrope is exposed before SRIF withdrawal is a major factor in determining the amplitude of GH bursts. SRIF (SRIF-14; a 30 micrograms/kg bolus, followed by 2 micrograms/kg.min) was infused iv for 40 min, GRF [GRF-(1-44) amide; 1 microgram/kg] was injected iv 20 min after starting the SRIF infusion, and the oGH rise after SRIF withdrawal was evaluated. In one series of controls GRF was replaced by saline, and in the other SRIF was replaced by saline. The oGH rise during recovery after SRIF alone was lower than that after the combined administration of SRIF and GRF (peak oGH increment, 8 +/- 3 vs. 38 +/- 12 ng/ml; P less than 0.04). The amplitude of the GH pulse after SRIF and GRF was similar to the immediate oGH response to GRF alone. These studies show that SRIF is unable to suppress the immediate oGH response to GRF in the ovine fetus, and that the suppressive effect of SRIF on the immediate oGH response to GRF increases gradually in late gestation and sharply at birth.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2563682 TI - The relative importance of nervous system and hormones to the 2-deoxy-D-glucose induced hyperglycemia in fed rats. AB - We examined the relative contributions of hormones and nervous system to the total 2-deoxy-D-glucose (2-DG)-induced central nervous system-mediated hyperglycemia. 2-DG was injected into the third cerebral ventricle in the following four groups of rats, and hepatic venous plasma glucose, immunoreactive glucagon, immunoreactive insulin, epinephrine, and norepinephrine were measured: 1) intact rats; 2) intact rats receiving somatostatin with insulin infusion through the femoral vein to inhibit glucagon secretion and maintain the basal insulin level; 3) bilateral adrenalectomized (ADX) rats to prevent epinephrine secretion; and 4) ADX rats receiving somatostatin with insulin infusion. Comparing areas under glucose curves among the intact rats, those receiving somatostatin with insulin infusion, ADX rats, and ADX rats receiving somatostatin with insulin infusion, the area under the glucose curve was intact rats greater than intact rats receiving somatostatin with insulin infusion greater than ADX rats receiving somatostatin with insulin infusion greater than ADX rats. These results suggest that there are three distinct sympathetic nervous system responses to 2-DG-induced central nervous system-mediated hyperglycemia. 2-DG induced hyperglycemia is not dependent on only one of those three systems, it is dependent on all of them. The relative potency of the factors to 2-DG-induced hyperglycemia increases in the following order: direct neural innervation of liver (including suppressive epinephrine action on insulin secretion), glucagon, and direct epinephrine action on liver. PMID- 2563683 TI - Paradoxical enhancement of pituitary growth hormone (GH) responsiveness to GH releasing factor in the face of high somatostatin tone. AB - Pituitary GH secretion is regulated by a delicate interplay between stimulatory (GRF) and inhibitory [somatostatin (SRIF)] hypothalamic hormones, although the nature of the GRF/SRIF interaction remains to be elucidated. In the present study, we documented a significant elevation of plasma SRIF-like immunoreactivity in 72-h fasted rats compared to that in fed controls (129.0 +/- 17.9 vs. 38.2 +/- 5.8 pg/ml; P less than 0.01) and used this model of high SRIF tone to further delineate the interrelation between GRF and SRIF in physiological regulation of pulsatile GH secretion. We examined pituitary GH responsiveness to GRF, both in vivo and in vitro, after 72-h exposure to nutritional deprivation and high SRIF secretion. In vivo, GRF-induced GH release was markedly enhanced in the face of high circulating SRIF; freely moving, starved rats released 4- to 8-fold more GH than fed controls in response to rat GRF iv. In vitro, both basal and human GRF induced GH release were augmented 2- to 4-fold in perifused dispersed anterior pituitary cells of starved rats compared to those in fed controls, and this enhanced responsiveness persisted in the presence of 10(-9) M SRIF. These results demonstrate that SRIF not only inhibits GH secretion stimulated by GRF, but that under different temporal conditions SRIF may act in a paradoxically positive manner to sensitize pituitary GH responsiveness to GRF. Such a cooperative interaction of the two peptides may be necessary to optimize pulsatile GH release. Our findings provide support for the hypothesis that the temporal patterning of hypothalamic GRF/SRIF signals to pituitary somatotrophs may be the major determinant for pulsatile GH secretion and, ultimately, body growth. PMID- 2563684 TI - Immunocytochemical and physiological evidence of a synapse between dopamine- and luteinizing hormone releasing hormone-containing neurons in the ewe median eminence. AB - Immunocytochemical labeling revealed that the arcuate nucleus (ARN) of the ewe's hypothalamus contains numerous tyrosine hydroxylase (TH)-positive neurons, that appear to lack dopamine-beta-hydroxylase (DBH)-like immunoreactivity. Axons of these presumed dopaminergic neurons converge in the median eminence (ME) with Luteinizing Hormone Releasing Hormone (LHRH)-containing axons originating mostly from neurons situated in the medial preoptic area. Electron microscopic double labeling revealed synaptic contacts between TH-positive presynaptic profiles and LHRH-containing postsynaptic elements. Samples of ME, ARN, paraarcuate and lateral hypothalamus were dissected and incubated to assess LHRH release and tissue content. Only ME-LHRH release was significantly reduced in the presence of dopamine (DA). All other regions released equal amounts with and without DA. Thus, a presynaptic dopaminergic inhibition of LHRH-containing axons at the level of the ME might contribute to the regulation of LHRH release into the portal vessels. PMID- 2563685 TI - Membrane phospholipid methylation is associated with surfactant secretion in rabbit type II alveolar epithelial cells. AB - We investigated the involvement of membrane phospholipid methylation in receptor mediated secretion of surfactant in adult rabbit type II alveolar epithelial cells (type II pneumocyte). Phospholipid methyltransferase activity was found in type II pneumocyte microsomes. Cell cultures of adult rabbit type II pneumocytes were then used to assay methyltransferase activity in the presence of the beta adrenergic agonist, terbutaline, and the methyltransferase inhibitor, 3 deazaadenosine. Terbutaline predictably stimulated adenylate cyclase activity and surfactant secretion. It was also found to stimulate incorporation of methyl groups into phosphatidylcholine and to increase beta-adrenergic receptor availability as assayed by binding of dihydroalprenolol (DHA). Surfactant secretion, as well as adenylate cyclase activity, were stimulated by terbutaline and were inhibited by 3-deazaadenosine. 3-Deazaadenosine did not inhibit DHA binding. These results suggest that phospholipid methylation plays a role in stimulus-secretion coupling in adult rabbit type II pneumocytes. PMID- 2563686 TI - Neurobiology of seizure predisposition in the genetically epilepsy-prone rat. AB - Seizure predisposition in the genetically epilepsy-prone rat (GEPR) is innately determined and these animals exhibit consistent and reproducible convulsive patterns. This epilepsy model is made up of 2 independently derived colonies of animals with each exhibiting a characteristic convulsive pattern. In response to a standardized acoustic stimulus, GEPR-3s exhibit moderate or clonic convulsions and GEPR-9s exhibit more severe tonic extensor convulsions. Besides exhibiting convulsions in response to sound stimulation, some GEPRs experience spontaneous and hyperthermic seizures. They are also abnormally sensitive to a number of seizure provoking stimuli that produce seizures in normal animals. The neurochemical basis for the seizure predisposition in GEPRs is increasingly well understood. Abnormalities in central nervous system norepinephrine and serotonin are widespread and may play a prominent role in regulation of seizures in the GEPR. Amino acid neurotransmitter systems are less well defined in the GEPR but abnormalities exist and may be, along with other documented deficiencies, responsible in part for the seizure predisposition that is characteristic of GEPRs. PMID- 2563687 TI - Induction of delayed synchronized bursts in hippocampal slices by weak sine-wave stimulation; role of the NMDA receptor. AB - Weak (20-50 microA) sine-wave stimulation at 60 Hz (SWS) of either the mossy fibers or the Schaffer collaterals promoted epileptiform synchronized bursts in the CA2/3 area of rat hippocampal slices in the absence of epileptogenic agents. Following brief SWSs (2-10 sec every 5 min), delayed synchronized bursts (DSBs) were triggered by weak test pulses in either pathway and transmitted to CA1. The long (2-10 sec) refractory periods which followed synchronized bursts in CA2/3 limited their rate of occurrence. Furthermore, SWS decreased the activity for several minutes in slices that exhibited frequent bursts. DSBs were reversibly blocked by perfusion with the N-methyl-D-aspartate (NMDA) specific antagonist DL 2-amino-5-phosphono-valeric acid (APV). The involvement of NMDA receptors was further suggested by the facilitation of CA2/3 synchronized bursts in medium with NMDA (5 microM) or lacking magnesium, and by iontophoresis of NMDA in the CA2/3 stratum radiatum. The findings that SWS-induced DSBs persisted for hours in undisturbed slices, and that bursts abolished by APV reappeared during washout in control solution, suggest long-term changes in the CA2/3 synaptic region. PMID- 2563688 TI - The efficacy of benorylate in postoperative dental pain. AB - The efficacy of a single pre-operative dose of benorylate (4 g) was determined in a double-blind, randomized, placebo-controlled parallel study in patients undergoing removal of a single impacted lower third molar. Patients treated with benorylate 4 g reported significantly less pain between 3-6 h after dosage than those treated with placebo. Overall pain scores at 6 h were significantly less in the benorylate group than the placebo group. However, overall pain scores at 12 h did not differ significantly between treatment groups. It is concluded from this study that a single dose of benorylate 4 g given immediately prior to the removal of an impacted lower third molar provides limited pain control during the postoperative period. PMID- 2563690 TI - Separation of CFU-S from primitive cells responsible for reconstitution of the bone marrow hemopoietic stem cell compartment following irradiation: evidence for a pre-CFU-S cell. AB - We have studied the in vivo spleen colony-forming ability and marrow repopulating ability of murine bone marrow cells differing in mitochondrial activity. Following centrifugal elutriation the cells were sorted on the basis of rhodamine 123 fluorescence intensity within a predetermined light scatter window. It is shown that a class of hemopoietic stem cells exists that differs from the majority of day-12 spleen colony-forming units (CFU-S) in that it has low mitochondrial activity and a high capacity to generate in time new day-12 CFU-S and cells that rescue recipients from radiation-inflicted death. These data add direct evidence for the identity of pre-CFU-S and CFU-S by physical separation. PMID- 2563691 TI - Immune thrombocytopenia in dogs after fetal liver hematopoietic cell transplantation. AB - Immune thrombocytopenia occurred in 6 of 33 engrafted dogs (18%) after fetal liver hematopoietic cell transplantation. Concurrent granulocytopenia occurred in three of six dogs and anemia in one. All dogs were receiving cyclosporin to prevent graft rejection and graft-versus-host disease (GVHD). None of the dogs had signs of GVHD. Bone marrow obtained at the time of platelet nadir was hypercellular with megakaryocyte hyperplasia. All dogs exhibited anti megakaryocyte antibodies detected by direct immunofluorescence of bone marrow smears. Treatment with oral prednisolone resulted in normalization of platelet counts in five of six dogs and granulocyte and erythrocyte counts in dogs exhibiting concurrent leukopenia or anemia. Two long-term survivors (greater than 2.5 years) have not developed further hematologic abnormalities since initial diagnosis and treatment. PMID- 2563689 TI - Debrisoquine oxidative phenotyping and psychiatric drug treatment. AB - The debrisoquine/sparteine phenotype was determined in 51 patients with depression, who were subdivided into 3 groups in terms of their drug treatment. Log (MR) for each group was compared. Patients treated with benzodiazepines had the same distribution of log (MR) as the healthy population, but the distribution was shifted towards higher values in patients treated with neuroleptics and antidepressants. It appears that the phenotypic expression of debrisoquine oxidation may be modified by drugs whose metabolism follows the same route as debrisoquine. The debrisoquine test must be carefully interpreted in patients receiving several drugs in the same time. PMID- 2563692 TI - Evaluation of a mouse Y chromosome probe for assessing marrow transplantation. AB - The multicopy mouse Y chromosome DNA probe 80Y/B (1) has been used to probe genomic DNA isolated from male and female bone marrow cells, mixed together in known ratios. It was found that in a mixture with a ratio of 1 male:200 female marrow cells the male cells could readily be detected by this method. The potential use of this technique for assessing repopulation kinetics of marrow transplantation is discussed. PMID- 2563693 TI - The influence of stereospecific assignments on the determination of three dimensional structures of proteins by nuclear magnetic resonance spectroscopy. Application to the sea anemone protein BDS-I. AB - The influence of the stereospecific assignments of beta-methylene protons and the classification of chi 1 torsion angles on the definition of the three-dimensional structures of proteins determined from NMR data is investigated using the sea anemone protein BDS-I (43 residues) as a model system. Two sets of structures are computed. The first set comprises 42 converged structures (denoted STEREO structures) calculated on the basis of the complete list of restraints derived from the NMR data, consisting of 489 interproton and 24 hydrogen bonding distance restraints, supplemented by 23 phi backbone and 21 chi 1 side chain torsion angle restraints. The second set comprises 31 converged structures (denoted NOSTEREO structures) calculated from a reduced data set in which those restraints arising from stereospecific assignments, and the corresponding chi 1 torsion angle restraints, are explicitly omitted. The results show that the inclusion of the stereospecific restraints leads to a significant improvement in the definition of the structure of BDS-I, both with respect to the backbone and the detailed arrangement of the side chains. Average atomic rms differences between the individual structures and the mean structures for the backbone atoms are 0.67 +/- 0.12 A and 0.93 +/- 0.16 A for the STEREO and NOSTEREO structures, respectively; the corresponding values for all atoms are 0.90 +/- 0.17 A and 1.17 +/- 0.17 A, respectively. In addition, while the overall fold remains unchanged, there is a small but significant atomic displacement between the two sets of structures. PMID- 2563694 TI - A liver-specific nuclear protein that binds to the distal promoter element of the rat tyrosine aminotransferase gene. AB - Using a gel retardation assay and exonuclease III footprinting, we have analyzed sequence-specific DNA-binding nuclear factors which interact with the distal promoter element of the rat tyrosine aminotransferase gene. A factor called LspA1, binding to a sequence that resembles the consensus binding site for the transcription factor Ap-1, was shown to be present in adult rat-liver nuclear protein extracts but not in the extracts from embryonic liver or spleen nuclei. PMID- 2563695 TI - Prophylactic short course rectal metronidazole for cesarean section. A double blind controlled trial of a simple low cost regimen. AB - One hundred and eighty two patients undergoing elective and emergency cesarean section were entered in a randomized double-blind placebo controlled trial of short course metronidazole rectal suppositories. There was a significant reduction in wound infection, febrile morbidity and postoperative hospitalization in the treated group. The reduction was greatest for serious wound infection. The simplicity and low cost of the regimen make it suitable for hospitals in developing countries. PMID- 2563696 TI - Uterine myoma in pregnancy: ultrasound study. AB - In many cases the presence of a myomatous formation in pregnancy begins its effect in a symptomless way and is revealed only by a regular sonographic control or at the time of delivery. In other cases, however, it could represent the cause of several complications, such as increased incidence of abortion extrauterine pregnancy premature breaking of the membranes, premature delivery distocia during delivery or it could itself be the site of necrotic processes. The attitude towards this type of pathology during pregnancy has often varied. The authors report their experiences regarding 408 cases of pregnancies complicated by myoma that were followed with accurate sonographic monitoring and they evaluate the incidence of the principal complications during pregnancy at the time of delivery and the eventual influence on the fetal weight at the time of birth. PMID- 2563697 TI - Characteristics of traditional midwives and their beliefs and practices in rural Bangladesh. AB - This study has analyzed the characteristics, beliefs and practices of midwives in rural Bangladesh. The midwives were mainly above age 30, married or widowed, and illiterate. Most of them learned their midwifery from informal sources such as female relatives or neighbours. Often, during pregnancy, childbirth, and post partum period, midwives imposed dietary restriction on the mothers. Similarly, devices used in the cutting of the umbilical cord and placenta were not properly sterilized and potentially dangerous substances were applied at the navel after cutting the umbilical cord or placenta. There was a practice of withholding breast-feeding up to 3 days after the birth of a child. However, there were also some beliefs or practices among the midwives that could be regarded as based on scientific understanding such as the practice of cutting the umbilical cord by boiled razor blade or the belief that child death could occur from tetanus caused by the unsterilized device used in the cutting of the umbilical cord. PMID- 2563698 TI - Cost effectiveness of ambulatory uterine activity monitoring. AB - A cost analysis is presented comparing 34 patients who received uterine activity monitoring versus 33 patients who attempted to detect uterine activity by palpation. All patients were at high risk for preterm delivery and were given the same educational information and prenatal care regarding signs and symptoms of preterm labor. The results revealed an increase in newborn days (640) and cost to those patients who were in the self-palpation group ($13,364) compared to monitored parturients (268 days and $8,633). The difference was attributed to neonatal morbidity from an increased number of preterm deliveries greater than 26 weeks but less than 37 weeks (P = 0.04). The increase in NICU days was significant (P = 0.03). No difference in normal newborn costs for infants delivered after greater than 33 weeks could be detected between the two groups, but morbidity was increased among control infants delivering between 34 and 36 weeks. Uterine activity monitoring to prevent preterm birth appears to be medically effective and reduces cost. PMID- 2563699 TI - Pelvic inflammatory disease and the intrauterine contraceptive device. AB - The relationship between the use of intrauterine contraceptive device (IUD) and pelvic inflammatory disease (PID) was examined in 1054 patients who were seen at the Family Planning Clinic of Jos University Teaching Hospital and were followed up. The overall risk of PID developing in women wearing the IUD was minimal: 62 out of 1054 (5.9%). The rates, however, varied for the 6-month periods studied. The incidence of PID decreased as the period of usage increased. The greater number of patients developed PID less than 3 months from the date of IUD insertion. When PID occurred it was usually of mild or moderate intensity and the response to antibiotic therapy was very encouraging. There was no relationship between the parity of the patients and the development of PID. Even though there is a definite link between the use of IUD and the development of PID it does not obliterate the benefits which the use of IUD provide for the majority of its patrons; and so the use of IUD should continue. PMID- 2563700 TI - Abdominal hysterectomies at area hospital, Point Fortin, Trinidad, West Indies. AB - A retrospective analysis of abdominal hysterectomies (3.08% of major operations) at the Area Hospital (52 beds), over a 5-year period revealed that the majority of the patients were multiparous in the reproductive age group. Menstrual irregularity is the most frequent indication, however, hysterectomy in these cases is not justified. High febrile morbidity (38%), blood transfusions and prolonged hospitalization were observed. Peer review, prophylactic antibiotics and survey of the Island hospitals are recommended to make hysterectomy appropriate, safe and to explain any racial variation. PMID- 2563701 TI - Hysterosalpingography versus laparoscopy in tubal infertility: comparison based on findings at laparatomy. AB - One hundred ten infertile women underwent hysterography (HSG) and laparoscopy at the Obafemi Awolowo University, Nigeria. Both techniques showed normal tubal patency in 62 (56.4%) women and abnormal pelvic pathology in 48 (43.6%). All 48 women underwent laparotomy for tuboplasty. At laparotomy, HSG and laparoscopic assessments were compared. Both techniques were comparable in the diagnosis of intratubal and distal tubal occlusion. However, laparoscopy was superior in the diagnosis of non-tubal factors and proximal tubal occlusion (P less than 0.002). It is suggested that laparoscopy should be the first procedure in the investigation of tubal infertility and only followed by HSG if the results show some abnormality. PMID- 2563702 TI - Culture of seminal fluid in infertile men and relationship to semen evaluation. AB - Bacterial flora of the seminal fluid and its influence on semen quality, was examined in 225 asymptomatic unselected men. Each semen sample was cultured aerobically, anaerobically, for genital mycoplasmas, and for Chlamydia trachomatis. Semen analysis was made according to standard methods recommended by the W.H.O. All 225 semen samples had microbial isolates. All isolates had colony counts of 10(2) colony forming units (cfu/ml). Thirty-three cases had greater than 10(2) cfu/ml, 85 cases had greater than 10(3) cfu/ml and 78 cases greater than 10(5) cfu/ml. The most common organisms isolated were Ureaplasma urealyticum in 86 samples and C. trachomatis in 26 samples. The most frequent abnormal parameters were viability (117 of 212, 52%), motility (85 of 212, 40%) and number of sperm cells (74 of 225, 32.8%). No significant correlation was found between abnormal semen parameters and presence of U. urealyticum, and C. trachomatis. We concluded that asymptomatic bacteriospermia (infection) in the semen did not significantly affect the count, motility or morphologic features of the specimen. PMID- 2563703 TI - Testicular volume and seminal fluid profile in fertile and infertile males in Ilorin, Nigeria. AB - Total testicular volume (TTV) measured with a metric tape and vernier calliper in 20 men whose wives were 12-20 weeks gestation, and in 48 infertile men was correlated with the height, weight, Ponderal Index (PI), sperm density, and total sperm in the ejaculate. There was a good correlation between TTV by tape and calliper (r = 0.96, P less than 0.001). There was a positive linear correlation between TTV and height but not with the other parameters. Giant head spermatozoa were more common in fertile Nigerian males than in fertile Asian or caucasian males (P less than 0.001). Giant head sperm and sperm with tail defects occurred in 6.25%, 11.1% fertile and 37.1%, 47.4% infertile men respectively. Spontaneous abortion due to blighted ova were more prevalent in wives whose husbands has greater than 8% giant head sperm (P less than 0.02). PMID- 2563704 TI - Importance of echographic and endocrine monitoring for the assessment of ovulation by follicle stimulating hormone in polycystic ovarian disease. AB - Sixteen patients with polycystic ovarian disease (PCOD) were treated for 39 cycles with pure follicle-stimulating hormone (FSH) for the induction of ovulation. At ovulation time human chorionic gonadotropin (hCG) was administered. Twenty-one cycles were ovulatory. Twenty-three were classified as normostimulated (N): six pregnancies and three abortions were observed. In the remaining eight hyperstimulated (H) cycles there were four full-term pregnancies. Dosage and length of treatment were greater in patients with excess body weight (P less than 0.01). H cycles were characterized in respect to N cycles by: (1) higher baseline values of 17-hydroxy progesterone (17-OHP) plasma levels and LH/FSH ratios; (2) higher plasma concentrations and rate of increase of 17-OHP periovulatory levels. E2 plasma levels did not permit a clear differentiation between H and N cycles, and it was not useful for the timely recognition of hyperstimulation. Our data show that a slight controlled degree of ovarian hyperstimulation is beneficial to pregnancy rate and outcome. PMID- 2563705 TI - Prostaglandin E2 induction of abortion and fetal demise. AB - Prostaglandin E2 (PGE2) suppositories have been shown to be active contractile agents and are effective in uterine evacuation for mid-trimester abortion or fetal demise. In this study, 85 patients were treated with vaginal PGE2 suppositories. When laminaria were used in patients with closed cervices, and compared to those who had minimal cervical dilatation, there was no difference in the time from induction to expulsion. Ninety-three percent of the 85 patients aborted successfully within 24 h. In each of the seven "failures", three or less suppositories were used prior to a dilatation and evacuation procedure. In this study, 81% of the abortions were complete, and in one-third of the remaining patients dilatation and curettage was performed just after delivery of the fetus. The incidence of minor side-effects ranged from 12 to 21%, and there were no major complications. It is concluded that the use of vaginal prostaglandin E2 suppositories for induction of mid-trimester abortion or fetal demise in the third trimester is safe and effective. PMID- 2563706 TI - Celo-intestinal fistulae complicating advanced extra-uterine pregnancy. AB - Multiple celo-intestinal fistulae with the passage of fetal bones per rectum as a rare complication of advanced extra-uterine pregnancy is presented. This was further complicated by a recent intra-uterine pregnancy which culminated in obstructed labor and uterine rupture. The subsequent peritonitis was of etiological significance in the formation of the peritoneo- or celo-intestinal fistulae. PMID- 2563707 TI - Acute polyhydramnios. AB - A case of acute polyhydramnios successfully managed via therapeutic amniocentesis is reported. A discussion and review of this rare entity is presented. PMID- 2563708 TI - Ogilvie's syndrome in the postcesarean section patient. AB - Two cases of colonic pseudo-obstruction (the so-called Ogilvie's syndrome) are reported. Both patients were in the immediate postcesarean section puerperium. The importance of early diagnosis in these cases is stressed, because this complication has a high mortality rate, frequently in relation to delayed diagnosis and treatment. One patient was successfully treated with conservative measures because diagnosis was made early. A plain X-ray abdominal film which shows cecum dilatation, with or without ascending and transverse colon dilatation, and no distal air, makes the diagnosis. A cecum diameter of 9 cm or more is a surgical indication, because the possibility of wall perforation is high. Surgical techniques are: puncture decompression or cecostomy. When cecum diameter is less than 9 cm, non-surgical measures (nasogastric suction, correction of any fluid and electrolytic imbalance, and maybe a flatus tube) are indicated. Observation through repeated X-ray abdominal films shows when the surgical indication appears: (1) failure of the conservative treatment (cecal distension continues or increases); or (2) cecal perforation is documented. PMID- 2563709 TI - AIDS. PMID- 2563710 TI - Transient catecholaminergic (TC) cells in the vagus nerves and bowel of fetal mice: relationship to the development of enteric neurons. AB - Catecholaminergic cells are transiently present during development of the fetal murine bowel. These transient catecholaminergic (TC) cells appear at Day E10, but by Day E13 can no longer be detected. In order to evaluate the hypothesis that these cells are the precursors of enteric neurons, we investigated the possibilities that TC cells coexpress neuronal and catecholaminergic markers, that they can be found along the presumed path followed by crest-derived cells migrating to the gut, and that they are proliferating. TC cells were identified immunocytochemically using polyclonal or monoclonal antibodies to tyrosine hydroxylase (TH). At Day E9.5, TH-immunoreactive cells were observed to be present along the wall of the primordial esophagus in lines that extended from the developing nodose ganglia down to the boundary of the stomach. At Day E9.5, TC cells were absent from the remaining foregut. These lines of esophageal TH immunoreactive cells became continuous with similar cells in the wall of the stomach and duodenum on Day E10. Coincident expression of neurofilament immunoreactivity was seen in all of the esophageal TH-immunoreactive cells present at Day E9.5, as well as in the entire set of esophageal and lower enteric TH-immunoreactive cells present at Day E10 (or later); moreover, at Days E9.5 and E10, all of the neurofilament-immunoreactive cells in the esophagus, stomach, or duodenum were also TH-immunoreactive. In contrast, neurofilament immunoreactivity was not expressed by the endodermally derived pancreatic duct and islet cells, which were also TH-immunoreactive; nor could expression of neurofilament immunoreactivity be detected in the TH-immunoreactive cells of the nodose ganglia. It was not until Day E11 that neurofilament-immunoreactive cells, which did not coexpress TH immunoreactivity (the definitive phenotype of enteric neurons) began to appear in the gut. Vagal axons reached as far distally as the nodose ganglion on Day E9.5, the esophagogastric junction on Day E10, and did not enter the stomach until Day E11. When the vagus nerves reached their level, the TH-immunoreactive cells in the wall of the esophagus came to lie among the nerve fibers. TH-immunoreactive cells are thus present on the pathway ultimately followed by the vagus nerves, but they develop before vagal fibers reach their level. The vagal TH-immunoreactive cells, therefore, are probably not initially migrating on vagal fibers, but appear instead to be overtaken by the descending vagus nerves.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2563711 TI - Effects of diabetes and diuresis on contraction and relaxation mechanisms in rat urinary bladder. AB - Experiments were designed to gain information on the mechanisms leading to diabetic urinary bladder dysfunction. Bladders from control rats, animals subjected to 4-5 wk streptozocin-induced diabetes, and rats subjected to equivalent diuresis produced by 5% sucrose feeding were studied with an in vitro whole-bladder preparation and neurochemical measurements. The diuretic group was used to distinguish alterations produced by metabolic effects on nerve and muscle from those induced by prolonged periods of excessive diuresis. Diuresis alone explains many of the diabetes-induced effects, including decreased norepinephrine levels, postsynaptic supersensitivity for sympathetic regulation of bladder storage, decreased responsiveness to parasympathetic regulation of emptying, and enhanced prostaglandin F2 alpha-induced contraction. Other diabetes-induced effects were not observed in the diuretic controls and are presumed to result from metabolic alterations associated with diabetes. These effects were decreases in norepinephrine uptake and in choline acetyltransferase activity, both markers of nerve terminal function. Thus, diuretic and metabolic factors appear to contribute to the early signs of parasympathetic and sympathetic neuropathy. In contrast, we found no evidence for loss of sensory nerve function in the diabetic bladder, at least at the organ level, because no diabetes- or diuresis-induced changes were observed in responsiveness to substance P or capsaicin. PMID- 2563712 TI - Increased neuropeptide Y concentrations in specific hypothalamic regions of streptozocin-induced diabetic rats. AB - Untreated streptozocin-induced diabetic (STZ-D) rats have previously been shown to have significantly increased hypothalamic concentrations of neuropeptide Y (NPY), a regulatory peptide that powerfully stimulates eating and drinking and inhibits secretion of several pituitary hormones when injected centrally. Tissue NPY concentrations have been measured by radioimmunoassay in selected hypothalamic regions microdissected from fresh, unfixed brain slices to localize diabetes-associated NPY changes precisely within the hypothalamus. Significant (35-200%) increases in NPY concentrations (P less than .01 vs. matched nondiabetic controls) were found in specific hypothalamic regions between 3 and 14 wk after induction of STZ-D. These regions included the paraventricular and ventromedial nuclei and lateral hypothalamic area, major appetite-regulating areas that are sensitive to the hyperphagic and polydipsic actions of NPY. Increased NPYergic activity in these areas may, at least partly, drive the increased eating and drinking characteristic of STZ-D. NPY concentrations were also increased in the arcuate nucleus and medial preoptic area. Because both of these regions are important in modulating pituitary hormone secretion, local NPY increases may be involved in the impaired secretion of luteinizing hormone, thyroid-stimulating hormone, growth hormone, and prolactin known to occur in STZ D. Our finding of NPY increases in specific hypothalamic nuclei associated with functional changes found in STZ-D suggests that this peptide may have a role in the altered metabolic and neuroendocrine regulation of the syndrome. PMID- 2563713 TI - Mapping mendelian factors underlying quantitative traits using RFLP linkage maps. AB - The advent of complete genetic linkage maps consisting of codominant DNA markers [typically restriction fragment length polymorphisms (RFLPs)] has stimulated interest in the systematic genetic dissection of discrete Mendelian factors underlying quantitative traits in experimental organisms. We describe here a set of analytical methods that modify and extend the classical theory for mapping such quantitative trait loci (QTLs). These include: (i) a method of identifying promising crosses for QTL mapping by exploiting a classical formula of SEWALL WRIGHT; (ii) a method (interval mapping) for exploiting the full power of RFLP linkage maps by adapting the approach of LOD score analysis used in human genetics, to obtain accurate estimates of the genetic location and phenotypic effect of QTLs; and (iii) a method (selective genotyping) that allows a substantial reduction in the number of progeny that need to be scored with the DNA markers. In addition to the exposition of the methods, explicit graphs are provided that allow experimental geneticists to estimate, in any particular case, the number of progeny required to map QTLs underlying a quantitative trait. PMID- 2563714 TI - Molecular genetic variation in the centromeric region of the X chromosome in three Drosophila ananassae populations. I. Contrasts between the vermilion and forked loci. AB - We have surveyed three natural populations of Drosophila ananassae for restriction map variation at the forked (f) and vermilion (v) loci, using 6 cutter restriction enzymes. Both loci are located in the centromeric region of the X chromosome. Two major conclusions can be drawn from the data. First, we found strong evidence for population subdivision, i.e., significant differences in the frequency distributions of polymorphisms and/or haplotypes between the Burma, India, and Brazil populations. Secondly, the pattern of DNA sequence variation between the two loci is unexpectedly different. The level of nucleotide variation in the v locus region is reduced (relative to f), especially in the Burma population. Furthermore, in contrast to v, we found no insertions/deletions larger than 700 bp and no significant linkage disequilibrium at f. The genetic differentiation among subpopulations can readily be attributed to restricted migration as the predominant evolutionary force. According to population genetics theory, the differences in DNA polymorphisms between the two loci are in qualitative agreement with the hypothesis that recombination is reduced in the v locus region ("centromere effect") but not at f. In order to test this hypothesis directly, we determined the cytogenetic positions of several loci in the centromeric region by in situ hybridization and found by comparison with the genetic map that recombination at v is indeed very low, much lower than at f. PMID- 2563715 TI - [German language reprints of original papers from the supplement "Nizatidine in peptic ulcer disease" of the Scandinavian Journal of Gastroenterology, Volume 22, Supplement 136, 1987]. PMID- 2563716 TI - [Alzheimer's disease. Review of the current status of research]. AB - The enormous social problems and costs caused by patients suffering from dementia induce growing public interest and become a great challenge of medical science. This report attempts to give a review of recent investigations in neuropathology, genetics, neurotransmitter research, epidemiology, diagnostics and therapy of Alzheimer's dementia, the most common type of dementia. A lot of recent molecular genetic experiments and many neuropathological analogies of Alzheimer's dementia and Down's syndrome indicate a damage on the chromosome 21 as possible cause of Alzheimer's dementia. The neuropathological changes are not limited to the grey matter and cholinergic system, but the white matter and some neurotransmitter systems (noradrenaline, dopamine, serotonin and somatostatin) are affected too. Therapeutical trials to compensate these transmitter deficits show no or only poor clinical benefit. Metabolic studies show disturbances in glucose metabolism of Alzheimer brains suggesting an intraneural energy deficit may be the main damage in Alzheimer's dementia. In spite of extensive technical and psychopathometrical diagnostical attempts Alzheimer's dementia remains to be difficult to diagnose precisely clinically. Best information is given by PET. PMID- 2563717 TI - Expression of type 1 fimbriae and mannose-sensitive hemagglutinin by recombinant plasmids. AB - Deletions within the cloned genes (fimA) encoding the type 1 major fimbrial subunits of two isolates of Klebsiella pneumoniae resulted in a nonfimbriate but hemagglutinating phenotype after transformation of Escherichia coli HB101 or ORN103. Phenotypic expression of type 1 fimbriae could be restored by transformation with plasmids containing the fimA genes of the fimbrial gene clusters from different strains. The surface fimbriae expressed were serologically identical to those of the polymerized product of the introduced fimA gene. The fimA gene products of Salmonella typhimurium and Serratia marcescens could utilize the accessory fimbrial genes of K. pneumoniae to produce surface-associated, hemagglutinating fimbriae. The relatedness of the type 1 fimbrial gene clusters from multiple isolates of members of the family Enterobacteriaceae was examined by DNA hybridization techniques. These analyses demonstrated little nucleotide sequence agreement among distinct genera of the enteric bacteria. PMID- 2563718 TI - Seroprevalence of HTLV-I in Portugal and evidence of double retrovirus infection of a healthy donor. AB - The prevalence of antibodies to HTLV-I in 5,475 Portuguese from 6 regions spanning the country was studied. Overall seroprevalence was 0.55%, indicating that Portugal is not an endemic area for this virus. Seropositives were distributed throughout the country, and no geographic clustering was observed. The seroprevalence of individuals who had lived in former Portuguese colonies in Africa (0.7%) was significantly higher than that of individuals who had not been in Africa (0.36%). An increase in seroprevalence with age was noted, and more females than males were antibody-positive, though not significantly so. Serum from one donor (1711), originating from Guinea-Bissau, was shown by Western blot and radioimmune precipitation to react with various proteins of HTLV-I, HIV-1 and -2, and SIV. Based on the serologic profiles and isolation of bona fide HTLV-I from her lymphocytes (confirmed by immunologic analysis, molecular cloning of the provirus and restriction enzyme analysis and sequencing of the DNA), together with the reactivity of her sera with an HIV-2 isolate obtained from her husband, we conclude that this donor was doubly infected with HTLV-I and HIV-2, rather than being the host to an as yet unidentified retrovirus. PMID- 2563719 TI - Structure and expression of c-erbB-2 and EGF receptor genes in inflammatory and non-inflammatory breast cancer: prognostic significance. AB - C-erbB-2 and epidermal growth factor receptor (EGFR) genes were independently shown to be associated with breast cancer progression. In this report, we have analyzed the structure and expression of these 2 genes in the same tumor specimens of a large series of breast cancers. Two clinical types of tumor were studied: inflammatory (IBC) and non-inflammatory breast cancers (NBC) obtained from 221 untreated patients at different clinical stages. Amplification and over expression of the c-erbB-2 proto-oncogene were observed in 27% and 47% of tumors, respectively, and were strongly associated with breast cancers of the most unfavorable prognosis, namely IBC and NBC with multiple positive axillary nodes. EGFR gene was neither amplified nor rearranged. A restriction fragment length polymorphism (RFLP) for HindIII endonuclease was observed. EGFR transcripts were detected in 46% of tumors and observed more frequently in IBC than in NBC (p less than 0.02). In NBC the presence of EGFR transcripts increased linearly with lymph node involvement and was associated with estrogen-receptor-negative tumors (p = 0.01). Analysis of both genes from the same tumor samples indicated that genes are associated with cancer aggressiveness. Furthermore, in NBC these 2 genes were independently activated, in contrast to IBC in which activated genes were negatively correlated, suggesting that c-erbB-2 and EGFR genes play different roles in NBC and IBC. PMID- 2563720 TI - The significance of oncogene amplification in primary breast cancer. AB - Alterations in the gene copy numbers of the proto-oncogenes HER2/neu and c-myc in primary human breast cancer investigated in 73 patients. We detected amplification of HER2/neu in 17 patient samples and amplification of c-myc in 11, while amplification of both was seen in 6 samples. There was no correlation of age, hormone receptor positivity or tumour size with amplification of either proto-oncogene. Amplification of HER2/neu was significantly correlated with the stage of the disease. HER2/neu amplification was observed in 18.5% and 38% of node-negative and node-positive patients, respectively; the association between HER2/neu amplification and advanced stage of the disease was statistically significant (p = 0.05). Since this is a prospective study, the clinical significance of oncogene amplification is not known. The relatively high frequency of HER2/neu amplification points to a functional role in human breast cancer, particularly in the progression of the disease. The method used in our study allows oncogene amplification to be studied in conjunction with hormone receptor determination and thus may be of value in large clinical trials to determine the significance of oncogene abnormalities in breast cancer. PMID- 2563721 TI - Characterization of HTLV-I isolates and T lymphoid cell lines derived from French West Indian patients with tropical spastic paraparesis. AB - Lymphoid cell lines derived from the peripheral blood of French West Indian patients with HTLV-I sero-positive Tropical Spastic Paraparesis and HTLV-I isolates were characterized. While patients' peripheral blood lymphocytes did not express detectable HTLV-I antigens when uncultured, they did so after short-term culture. Established cell lines were of T-cell lineage: CD2+, CD3+, CD4+, CD7+, WT31+ with activated T-cell markers CD25+, DR+ and a clonal rearrangement of the beta and gamma genes of the T-cell receptor. HTLV-I antigens were detected in cell lines by indirect immunofluorescence, Western blot and radio immunoprecipitation assays. After 4 months in culture, low levels of Mg2+ dependent reverse transcriptase activity were detected and electron microscopy revealed numerous type-C retroviral particles similar to HTLV-I virions. Western blot and radio-immunoprecipitation analysis of purified viruses revealed gp46, p24, p19 and Pr53gag proteins similar to those detected in HUT 102 and MT2 cell lines. Deep analysis of env-coded precursor of one TSP versus ATL isolates revealed minor differences in their molecular weights. Southern blot analysis using 32P HTLV-I env gene as a probe showed the presence of HTLV-I proviral fragments clonally integrated into the genome of the cell lines. Our data suggest that HTLV-I isolated from Tropical Spastic Paraparesis does not differ significantly from the leukemogenic prototypes. Does HTLV-I induce either acute lymphoproliferative diseases or chronic neuromyelopathies depending upon as yet unknown co-factors? This question remains to be determined. PMID- 2563722 TI - Immunohistochemical detection of P-glycoprotein in human tumor cells with a low degree of drug resistance. AB - We report the immunohistochemical detection of the 170-180 kDa multi-drug resistance-related P-glycoprotein in human tumor cells with a low level of resistance. A series of human squamous lung cancer cell lines with increasing levels of resistance to doxorubicin (DOX) was developed and stained for P glycoprotein, using the JSB-IMAb. Subline SW1573/50A with a 4- to 6-fold cross resistance to daunorubicin (DNR) and vincristine (VCR) showed rather uniform positive staining for P-glycoprotein apparently at cytoplasmic sites. Only in cells with higher degrees of resistance (greater than 10-fold) could plasma membrane-associated P-glycoprotein be made visible. DNR efflux was increased in SW1573/50A as compared to the parent line SW1573 (52 and 70% DNR were retained during 3 min efflux respectively). Verapamil partially reversed DNR and VCR resistance in SW1573/50A. Cells obtained from a metastasized renal cell carcinoma and cultured in vitro stained in a similar way to SW1573/50A and showed some sensitivity to verapamil modulation of VCR cytotoxicity. Our results suggest that weakly resistant cancer cells obtained from patients can be routinely detected with JSB-I on cytospins, and implicate that in such weakly resistant cells P glycoprotein may be present, while plasma membrane expression is not yet readily detectable. PMID- 2563724 TI - Isolation of enterotoxigenic Escherichia coli from a foal with diarrhea. AB - Enterotoxigenic Escherichia coli was isolated from a 3-day-old foal with diarrhea. The isolate was distinguished from nonpathogenic E coli by determining the presence of pili and enterotoxin production. A standard slide agglutination test was performed, using pooled antisera that contained antibodies against K99 and F41 pilus antigens, K87 capsular antigen, and 0101 somatic antigen. Agglutination of the antisera occurred in the presence of the isolate. Piliation was verified by use of negative-contrast electron microscopy. Further, the isolate produced a heat-labile enterotoxin-like antigen that cross-reacted with a reagent containing formalin-treated, heat-killed Staphylococcus aureus (cowan 1 strain) bearing anti-cholera antibodies. On the basis of the aforementioned procedures and the absence of other identifiable enteric pathogens, we believe that E coli was responsible for causing diarrhea in the foal. PMID- 2563725 TI - Daily exercise enhances coronary resistance vessel sensitivity to pharmacological activation. AB - The effect of daily exercise on the coronary resistance vessel sensitivity to intracoronary infusion of several pharmacological agents was assessed in 12 conscious adult mongrel dogs. alpha-Adrenergic receptor agonists (norepinephrine and phenylephrine) significantly decreased coronary blood flow velocity. beta 2 Adrenergic receptor agonists (isoproterenol and zinterol) and a metabolic vasodilator (adenosine) significantly increased coronary blood flow velocity. These responses occurred without altering factors that influence myocardial metabolism. Daily exercise significantly enhanced the coronary vascular sensitivity to each of the pharmacological agents. These results suggest that a nonspecific potentiation to pharmacological activation occurs after daily exercise. After left stellate ganglionectomy, intracoronary infusions of each pharmacological agent had similar effects on coronary blood flow velocity as presented for the intact dogs; however, daily exercise did not enhance the coronary vascular sensitivity to the pharmacological agents. These results demonstrate the need for an intact nervous system for the vascular adaptations associated with daily exercise. PMID- 2563723 TI - Generalized lichen nitidus. Report of two cases treated with astemizol. AB - Two patients, one man aged 65 and one woman aged 48, presenting generalized lichen nitidus are reported. The clinical and histopathologic features of this uncommon presentation of lichen nitidus are illustrated as well as the excellent clinical response to the treatment with an H1-blocking antihistaminic (astemizol). PMID- 2563726 TI - Adrenergic stimulation of inositol phosphate accumulation in tracheal epithelium. AB - To determine whether inositol phosphates are important second messengers in the regulation of Cl- secretion by airway epithelia, we examined the relationship between inositol phosphate accumulation and Cl- secretion in response to adrenergic agonists. We found that epinephrine stimulated Cl- secretion and inositol phosphate accumulation with similar concentration dependence. Although isoproterenol stimulated Cl- secretion, there was no effect of beta-adrenergic receptor activation on inositol phosphate accumulation. In contrast, alpha 1 adrenergic receptor activation stimulated inositol phosphate accumulation but failed to induce Cl- secretion. Another Cl- secretagogue, prostaglandin E1, also failed to stimulate inositol phosphate accumulation. These data suggest that inositol phosphate accumulation is neither sufficient nor required for stimulation of Cl- secretion in cultured canine tracheal epithelial cells. PMID- 2563727 TI - Transcriptional regulation of proto-oncogene expression by epidermal growth factor, transforming growth factor beta 1, and triiodothyronine in MDA-468 cells. AB - We have examined the epidermal growth factor (EGF) dependence of the transcription of different proto-oncogenes, using cultured human mammary carcinoma MDA-468 cells and a nuclear run-on transcription assay. We found that the stimulation of MDA-468 cells with EGF regulates moderately and to different extents the transcription of the EGF-receptor(R) and c-erbB-2 proto-oncogenes. In contrast, the transcription of other proto-oncogenes, including c-myc, c-H-ras, and c-fps, was unchanged. Furthermore, we provide evidence that transforming growth factor beta 1 (TGF-beta 1) selectively and to different degrees modulates the EGF-dependent transcription of EGF-R and c-erbB-2 genes. In this study, we also discovered that T3 (triiodothyronine) exerts synergistic control on the action of EGF alone or in association with TGF-beta 1, on EGF-R and c-erbB-2 gene transcription. Moreover, we established that within 6 h after the addition of EGF, cytoplasmic EGF-R mRNA levels are increased several-fold and that this accumulation is enhanced by the presence of TGF-beta 1 and/or T3. The results described here are consistent with the hypothesis that a complex program of cooperative interactions among EGF-, TGF-beta 1-, and T3-generated signals at the transcriptional level may mediate, at least in part, the combined actions of EGF, TGF-beta 1, and T3 in target cells. PMID- 2563728 TI - Hypertonic media inhibit receptor-mediated endocytosis by blocking clathrin coated pit formation. AB - Two seemingly unrelated experimental treatments inhibit receptor mediated endocytosis: (a) depletion of intracellular K+ (Larkin, J. M., M. S. Brown, J. L. Goldstein, and R. G. W. Anderson. 1983. Cell. 33:273-285); and (b) treatment with hypertonic media (Daukas, G., and S. H. Zigmond. 1985. J. Cell Biol. 101:1673 1679). Since the former inhibits the formation of clathrin-coated pits (Larkin, J. M., W. D. Donzell, and R. G. W. Anderson, 1986. J. Cell Biol. 103:2619-2627), we were interested in determining whether hypertonic treatment has the same effect, and if so, why. Fibroblasts (human or chicken) were incubated in normal saline made hypertonic with 0.45 M sucrose, then broken open by sonication and freeze-etched to generate replicas of their inner membrane surfaces. Whereas untreated cells display typical geodesic lattices of clathrin under each coated pit, hypertonic cells display in addition a number of empty clathrin "microcages". At first, these appear around the edges of normal coated pit lattices. With further time in hypertonic medium, however, normal lattices largely disappear and are replaced by accumulations of microcages. Concomitantly, low density lipoprotein (LDL) receptors lose their normal clustered distribution and become dispersed all over the cell surface, as seen by fluorescence microscopy and freeze-etch electron microscopy of LDL attached to the cell surface. Upon return to normal medium at 37 degrees C, these changes promptly reverse. Within 2 min, small clusters of LDL reappear on the surfaces of cells and normal clathrin lattices begin to reappear inside; the size and number of these receptor/clathrin complexes returns to normal over the next 10 min. Thus, in spite of their seeming unrelatedness, both K+ depletion and hypertonic treatment cause coated pits to disappear, and both induce abnormal clathrin polymerization into empty microcages. This suggests that in both cases, an abnormal formation of microcages inhibits endocytosis by rendering clathrin unavailable for assembly into normal coated pits. PMID- 2563730 TI - Effects of the somatostatin analog BIM 23014 on the secretion of growth hormone, thyrotropin, and digestive peptides in normal men. AB - BIM 23014 (BIM) is a long-acting octapeptide somatostatin analog. We studied the effects of this analog on the secretion of GH, TSH, and gastroenteropancreatic hormones [secretin, motilin, and pancreatic polypeptide (PP)] in normal men. In the first protocol three BIM doses (125, 250, and 500 micrograms) and vehicle were administered sc in random order at 2000 h to eight normal young men. Plasma GH concentrations decreased during the first part of the night only after the highest dose (P less than 0.05). Plasma secretin levels did not change, while plasma motilin decreased after the 250- and 500-micrograms doses (P = 0.05 and P = 0.02, respectively), and plasma PP decreased after all three doses (P less than 0.05, P less than 0.01, and P less than 0.01, respectively) during the first part of the night. In the second protocol, eight men received BIM, administered by constant sc infusion during the night in a dose of 2 mg/12 h, or vehicle, either alone or in association with a 10 ng/kg.min iv GHRH or vehicle infusion. Nocturnal GH secretion was suppressed by the BIM infusion (P less than 0.001). GH secretion, stimulated by GHRH infusion (P less than 0.001), was reduced by concomitant BIM infusion (P less than 0.001) and was pulsatile during the combined infusions. BIM infusion suppressed the physiological nighttime rise in plasma TSH levels. Plasma motilin and PP levels were reduced by BIM, when administered either alone or in combination with GHRH. We conclude that: 1) BIM is capable of reducing GH secretion when administered sc in a dose of 500 micrograms and of abolishing nocturnal GH secretion when constantly infused at a dose of 2 mg/12 h; 2) BIM, constantly infused, reduces the nocturnal rise in TSH secretion; and 3) motilin and PP secretion are more sensitive than that of GH to BIM, as they are reduced by a lower dose. PMID- 2563729 TI - Effects of cytoplasmic acidification on clathrin lattice morphology. AB - Reducing the internal pH of cultured cells by several different protocols that block endocytosis is found to alter the structure of clathrin lattices on the inside of the plasma membrane. Lattices curve inward until they become almost spherical yet remain stubbornly attached to the membrane. Also, the lattices bloom empty "microcages" of clathrin around their edges. Correspondingly, broken open cells bathed in acidified media demonstrate similar changes in clathrin lattices. Acidification accentuates the normal tendency of lattices to round up in vitro and also stimulates them to nucleate microcage formation from pure solutions of clathrin. On the other hand, several conditions that also inhibit endocytosis have been found to create, instead of unusually curved clathrin lattices with extraneous microcages, a preponderance of unusually flat lattices. These treatments include pH-"clamping" cells at neutrality with nigericin, swelling cells with hypotonic media, and sticking cells to the surface of a culture dish with soluble polylysine. Again, the unusually flat lattices in such cells display a tendency to round up and to nucleate clathrin microcage formation during subsequent in vitro acidification. This indicates that regardless of the initial curvature of clathrin lattices, they all display an ability to grow and increase their curvature in vitro, and this is enhanced by lowering ambient pH. Possibly, clathrin lattice growth and curvature in vivo may also be stimulated by a local drop in pH around clusters of membrane receptors. PMID- 2563731 TI - Plasma dihydroxyphenylalanine and total body and regional noradrenergic activity in humans. AB - Dihydroxyphenylalanine (DOPA) is the immediate product of the rate-limiting step in catecholamine biosynthesis, hydroxylation of tyrosine. This study examined whether plasma concentrations of DOPA are related to tyrosine hydroxylase activity. Plasma concentrations of DOPA, norepinephrine, and the norepinephrine metabolites 3,4-dihydroxyphenylglycol (DHPG) and 3-methoxy-4-hydroxyphenylglycol (MHPG) were measured in arterial blood and blood draining the heart, brain, and forearm of 21 patients undergoing cardiac catheterization. Rates of entry of norepinephrine into arterial plasma and plasma draining the heart were estimated using infusions of radioactive norepinephrine. Arterial plasma DOPA correlated positively with arterial plasma DHPG (r = 0.63), MHPG (r = 0.47), norepinephrine (r = 0.67), and the rate of entry of norepinephrine into arterial plasma (r = 0.62). There were significant arteriovenous increments in plasma DOPA: 28% across the heart, 18% across the brain, and 32% across the forearm. Arteriovenous increments in plasma DOPA across the brain correlated positively with increments in plasma DHPG (r = 0.83), but not with increments in norepinephrine or MHPG. In the arm, where MHPG was the major metabolite, arteriovenous increments in DOPA correlated positively with increments in MHPG (r = 0.52) and with the combined increments in MHPG, DHPG, and norepinephrine (r = 0.60). In the heart, where DHPG was the major metabolite, arteriovenous increments in DOPA correlated positively with increments in DHPG (r = 0.72) and the combined increments in DHPG, MHPG, and norepinephrine (r = 0.62). The rate at which norepinephrine entered the great cardiac venous plasma from tissues of the heart correlated positively with the rate at which DOPA overflowed from the heart into the systemic circulation (r = 0.56). The relationships between plasma DOPA and norepinephrine metabolism and the rates of norepinephrine entry into plasma support the view that plasma DOPA reflects tyrosine hydroxylase activity. PMID- 2563732 TI - Influence of growth hormone on glucose-induced glucose uptake in normal men as assessed by the hyperglycemic clamp technique. AB - To determine whether physiological increments in circulating GH concentrations influence glucose-induced glucose uptake (GIGU), two-step sequential hyperglycemic clamp (plasma glucose, 6 and 14 mmol/L) studies were performed in six normal subjects with and without GH infusion (40 ng/kg.min). The latter resulted in serum GH levels of 15 +/- 1 (+/- SE) microgram/L. Infusion of somatostatin (250 micrograms/h during step 1 and 750 micrograms/h during step 2) together with a replacement dose of insulin (1.1 pmol/kg.min) resulted in serum insulin levels comparable to basal levels in both studies. The GIGU ([3 3H]glucose), assessed as the difference between steps 2 and 1 glucose utilization during the final 60 min of each step (150 min) was markedly impaired during GH infusion (with GH, 1.1 +/- 0.2 mg/kg.min; without GH, 3.1 +/- 0.3 mg/kg.min; P less than 0.001). Moreover, the percent increase in glucose uptake was considerably reduced during hypersomatotropinemia (with GH, 44 +/- 9%; without GH, 97 +/- 11%; P less than 0.01). In the GH infusion as well as control studies, endogenous glucose production (EGP) was similar at the two levels of glycemia, whereas GH infusion approximately doubled EGP [2.3 +/- 0.2 vs. 1.1 +/- 0.3 mg/kg.min and 2.0 +/- 0.4 vs. 1.1 +/- 0.4 mg/kg.min (step 1 and 2, respectively)]. We conclude that moderate hypersomatotropinemia for several hours is characterized by impaired GIGU as well as augmented EGP. PMID- 2563733 TI - Specific effect of CV 205-502, a potent nonergot dopamine agonist, during a combined anterior pituitary function test. AB - CV 205-502, an octahydrobenzo[g]quinoline, is a dopamine agonist compound that is not an ergot or ergoline derivative. To investigate the site of action of CV 205 502, three groups of five men were given single daily doses of CV 205-502 (0.04, 0.06, or 0.08 mg/day, doses that suppress plasma PRL by 60-80% for 24 h) for 5 days; on day 6 a combined anterior pituitary function test using iv administration of four hypothalamic releasing hormones (TRH, 200 micrograms; GHRH, 100 micrograms; CRH, 100 micrograms; LHRH, 100 micrograms) was performed. One month later the challenge tests were repeated to obtain control values. The following hormones were measured by RIA in plasma: TSH, ACTH, cortisol, PRL, GH, LH, FSH, and testosterone. With the exception of plasma PRL levels, basal and releasing hormone-stimulated values were similar after CV 205-502 administration and after the 1-month washout period. Basal plasma PRL was lower after CV 205-502 administration, and the response to TRH was attenuated by all three doses of CV 205-502 (the mean percent inhibition values were 76%, 93%, and 94%, respectively). All three doses of CV 205-502 were well tolerated, and another group of men well tolerated 0.1 mg daily. The results confirm that CV 205-502 is a potent dopamine agonist, which directly inhibits lactotropic cells but has no effect on other pituitary cell types. PMID- 2563734 TI - Half-time of endogenous growth hormone (GH) disappearance in normal man after stimulation of GH secretion by GH-releasing hormone and suppression with somatostatin. AB - The half-life (t1/2) of disappearance of endogenous GH from serum was studied using physiological effectors to stimulate and then suppress GH release. GH secretion was stimulated by a single iv injection of GHRH, followed 45 min later by an iv bolus dose and then a 2.5-h infusion of somatostatin (SRIH) to suppress further release. The in vivo t1/2 of GH in seven men was calculated from serum GH concentrations measured at frequent intervals after beginning the SRIH infusion. The mean t1/2 of endogenous GH was 18.9 +/- 0.8 (+/- SE) min by monoexponential analysis and 3.5 +/- 0.7 and 20.7 +/- 0.7 min by biexponential fitting. In these normal men, the decline in GH concentrations after GHRH and SRIH administration was similar to that after the administration of GHRH alone, which yielded a t1/2 of 20.3 +/- 1.9 min. We conclude that the physiological kinetics of endogenous GH removal/disappearance can be estimated in vivo in man using GHRH with or without SRIH infusion. PMID- 2563735 TI - Impairment of noninsulin-mediated glucose disposal in the elderly. AB - While normal aging is characterized by resistance to insulin-mediated glucose disposal (IMGU), the effect of age on noninsulin-mediated glucose disposal (NIMGU), which is responsible for the majority of basal glucose uptake, has not been completely evaluated. These studies were conducted on healthy nonobese young (n = 10; age, 20-30 yr) and old (n = 10; age, 62-80 yr) men. Each subject underwent two paired studies in random order. In all studies a [3H]glucose infusion was used to measure glucose uptake and production rates, and somatostatin (500 micrograms/h) was infused to suppress endogenous insulin release. In study A, plasma glucose was kept close to fasting levels (approximately 5.6 mmol/L) using an euglycemic clamp protocol for 4 h. Plasma insulin decreased to less than 20 pmol/L within 15 min and remained suppressed thereafter in all studies. Steady state (15-240 min) plasma glucagon levels were slightly greater in the elderly [young, 86 +/- 5 (+/- SE); old, 98 +/- 2 ng/L; P less than .05]. Basal glucose uptake was similar in both groups (young, 877 +/- 21; old, 901 +/- 24 mumol/min). Glucose uptake during the last hour of the study (180-240 min) was used to represent NIMGU, because insulin action was assumed to be absent by this time. NIMGU was less in the elderly (young, 744 +/- 18; old, 632 +/- 32 mumol/min; P less than 0.01). In study B, plasma glucose was kept at about 11 mmol/L for 4 h using a hyperglycemic clamp protocol. Plasma insulin decreased to less than 20 pmol/L within 15 min and remained suppressed thereafter in all studies. Steady state plasma glucagon levels were slightly but not significantly higher in the elderly (young, 88 +/- 6; old, 100 +/- 4 ng/L). Basal glucose uptake (young, 910 +/- 27; old, 883 +/- 25 mumol/min) and NIMGU (young, 933 +/- 36; old, 890 +/- 16 mumol/min; P = NS) were similar in both young and old subjects. We conclude that aging is associated with impairment in NIMGU only in the basal state, which may explain in part the increase in fasting glucose with age. PMID- 2563736 TI - Inhibition of lipopolysaccharide-induced alpha/beta-interferon synthesis by dopamine and the dopamine-1 agonist fenoldopam. AB - In vitro incubation of Listeria monocytogenes-immune spleen cells in the presence of dopamine or fenoldopam, a dopamine-1 (D1) agonist, inhibited alpha/beta interferon (IFN) synthesis induced by the mitogen lipopolysaccharide (LPS), in a manner that appeared to be concentration dependent. In addition, the inhibitory effect of dopamine and fenoldopam on the synthesis of IFN was prevented by incubating immune spleen cells in the presence of haloperidol, a D1 antagonist. PMID- 2563737 TI - Ventral mesencephalic neurons containing both cholecystokinin- and tyrosine hydroxylase-like immunoreactivities project to forebrain regions. AB - The coexistence of cholecystokinin- and tyrosine hydroxylase-like immunoreactivities within neurons of the rat ventral mesencephalon was analyzed by using an indirect immunofluorescence technique for the simultaneous demonstration of two antigens in the same tissue section. A high degree of colocalization was observed in the substantia nigra pars compacta, in which 80 90% of all labeled neurons at rostral and up to 70% at intermediate levels contained both cholecystokinin and tyrosine hydroxylase. At caudal levels, the incidence of colocalization declined to approximately 30-50%. All of the immunoreactive perikarya in the substantia nigra pars lateralis were labeled with both substances. Other areas of the ventral midbrain that exhibited a moderate proportion of neurons immunoreactive for both cholecystokinin and tyrosine hydroxylase included the ventral tegmental area, interfascicular nucleus, and rostral and caudal linear nuclei. In addition, coexistence was occasionally observed within neurons of the central and ventral periaqueductal gray matter, supramammillary region, peripeduncular region, retrorubral field, and extremely rarely, within the substantia nigra pars reticulata. Cell bodies containing tyrosine hydroxylase-like immunoreactivity (indicative of dopamine) usually outnumbered those containing the peptide except in the supramammillary region and in the ventral periaqueductal gray matter, where the cholecystokinin perikarya were present in higher numbers. The double-labeling colocalization technique was combined with fluorescence retrograde tracing to determine some of the forebrain projections of these neurons. Ventral midbrain neurons containing both cholecystokinin and tyrosine hydroxylase were found to project to the caudate putamen, nucleus-accumbens, prefrontal cortex, and amygdala. These projections originated from neurons located predominantly in the substantia nigra pars compacta and the ventral tegmental area. Thus, cholecystokinin occurs within the well-known dopaminergic nigrostriatal pathway in the rat. Overall, these results demonstrate that a significant proportion of the dopamine neurons giving rise to the ascending mesotelencephalic projections also contain the peptide cholecystokinin. PMID- 2563738 TI - Comparison of met-enkephalin, dynorphin A, and neurotensin immunoreactive neurons in the cat and rat spinal cords: II. Segmental differences in the marginal zone. AB - This study examined the number of met-enkephalin, dynorphin A 1-8, and neurotensin immunoreactive (IR) neurons in the marginal zone (lamina I) at one thoracic (T8:cat,T9:rat), one midlumbar (L5:cat,L4:rat), and one lower lumbar or sacral (S1:cat,L6:rat) spinal cord segment in the cat and rat. Marginal zone IR neurons ranged 10-70 microns in diameter in cats and 10-50 microns in rats and were flattened, pyramidal, fusiform, or polygonal in morphology. Immunoreactive neurons for each peptide in both species were found in the marginal zone at all spinal levels, but with a differential segmental distribution. The average number of IR neurons per 50-microns section generally was lowest in thoracic cord and greatest in lower lumbar/sacral cord for all peptides. For enkephalin and dynorphin, the estimated total number of IR neurons per segment and number of IR neurons per volume (mm3) generally were lowest in the midlumbar segments and highest in the thoracic and lower lumbar/sacral cord. For neurotensin, the estimated total number of neurons per segment remained lowest in the thoracic and largest in the lower lumbar/sacral cord. The number of neurotensin IR neurons per volume was equal in the thoracic and midlumbar cord, but remained highest at lower lumbar/sacral levels. The IR neurons quantified in this study may be interneurons or may serve as supraspinal projection neurons. The large number of IR neurons observed in segments receiving a relatively large visceral afferent input suggests that some of these neurons may be involved in visceral sensory processing. In addition, the segmental distribution of the IR neurons indicates that physiological and pharmacological studies on the effects of opioid and/or neurotensin peptides should be interpreted in light of the spinal segment(s) investigated. PMID- 2563739 TI - Immunohistochemical localization of seven different peptides in the human spinal cord. AB - It is necessary to study the normal chemical contents in the human spinal cord in order to understand neurochemical changes that might occur under pathological conditions. In the present study, the comparative distribution of seven peptides was examined immunohistochemically in four levels (cervical, C; thoracic, T; lumbar, L; sacral, S) of the human spinal cord by means of the peroxidase antiperoxidase technique. The peptides examined included bombesin (BOM), substance P (SP), cholecystokinin (CCK), somatostatin (SOM), methionine enkephalin (M-ENK), vasoactive intestinal polypeptide (VIP), and thyrotropin releasing hormone (TRH). Among the seven peptides examined, four (BOM, CCK, SOM, and TRH) have never been described in the human spinal cord and the present work clearly demonstrates their existence in specific patterns. The terminals that were immunostained for BOM and CCK were localized in high concentration in the superficial dorsal horn (laminae I-II), in moderate amounts in the lateral part of laminae V and VII, and lesser amounts in the intermediate gray (lamina VII) and the dorsal part of the central gray (lamina X). Whereas BOM showed a similar distribution pattern at all spinal levels, CCK was mainly found in thoracic and lumbar levels. The SOM terminals were localized in the superficial dorsal horn (the highest density in lamina II but very few in lamina I), the intermediolateral cell column, intermediate gray, and central gray. This peptide was more widely distributed in the sacral cord with its terminal field extending into the ventral horn. The TRH terminals were mainly located in the ventral horn. Frequently, TRH terminals were seen adjacent to large ventral horn neurons. Furthermore, many neurons in the ventral and intermediate gray and Clarke's column demonstrated TRH immunoreactivity. The other three peptides (SP, M-ENK, and VIP) have been previously demonstrated in the human spinal cord and the present study confirmed their general spinal distribution with minor differences. PMID- 2563740 TI - Distribution of somatostatin receptors in the brain of the frog Rana ridibunda: correlation with the localization of somatostatin-containing neurons. AB - The biochemical characterization and anatomical distribution of somatostatin binding sites were examined in the brain of the frog Rana ridibunda, and the distribution of the receptors was compared with the location of somatostatin immunoreactive neurons. The pharmacological profile of somatostatin receptors was determined in the frog brain by means of an iodinated superagonist of somatostatin, [125I-Tyr0,DTrp8]S-14. Membrane-enriched preparations from frog brain homogenates were shown to contain high-affinity receptors (KD = 0.78 +/- 0.34 nM; Bmax = 103 + 12.7 fmoles/mg protein) with pharmacological specificity for [DTrp] substituted S14 and S28 analogs. The distribution of somatostatin binding sites was studied by autoradiography on coronal sections of frog brain. Various densities of somatostatin receptors were detected in discrete areas of the brain. The highest concentration of binding sites was observed in the olfactory bulb, in the pallium, and in the superficial tectum. Moderate binding was observed in the striatum, amygdaloid complex, preoptic area, and cerebellum. Immunocytochemical studies of the distribution of somatostatin-28 (S28) related peptides were also conducted in the frog brain. Two antisera that recognize distinct epitopes of the somatostatin molecule have been used for immunohistochemical mapping of the peptide. Antiserum SS9 recognizes both S28 and somatostatin-14 (S14) and allowed the labelling of perikarya. Antiserum S320 recognizes the N-terminal fragment (1-12) resulting from enzymatic cleavage of S28. This latter antiserum, which does not cross-react with S28, stained mainly neuronal processes. At the infundibular level, however, both antisera stained cell bodies and fibers. Immunoreactive somatostatin-related peptides were detected in many areas of the frog brain. In the diencephalon, a heavy accumulation of perikarya and fibers was seen in the preoptic nucleus, the dorsal and ventral infundibular nuclei, and the median eminence. Immunoreactive perikarya were also observed in the telencephalon, especially in the pallium and in thalamic nuclei. Immunostained processes were detected in many telencephalic areas and in the tectum. There was good correlation between the distribution of somatostatin-immunoreactive elements and the location of somatostatin-binding sites in several areas of the brain, in particular in the median pallium, the tectum, and the interpeduncular nucleus.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2563741 TI - Improved diagnostic performance of exercise thallium-201 single photon emission computed tomography over planar imaging in the diagnosis of coronary artery disease: a receiver operating characteristic analysis. AB - Qualitative interpretation of tomographic and planar scintigrams, a five point rating scale and receiver operating characteristic analysis were utilized to compare single photon emission computed tomography and conventional planar imaging of myocardial thallium-201 uptake in the accuracy of the diagnosis of coronary artery disease and individual vessel involvement. One hundred twelve patients undergoing cardiac catheterization and 23 normal volunteers performed symptom-limited treadmill exercise, followed by stress and redistribution imaging by both tomographic and planar techniques, with the order determined randomly. Paired receiver operating characteristic curves revealed that single photon emission computed tomography was more accurate than planar imaging over the entire range of decision thresholds for the overall detection and exclusion of coronary artery disease and involvement of the left anterior descending and left circumflex coronary arteries. Tomography offered relatively greater advantages in male patients and in patients with milder forms of coronary artery disease, who had no prior myocardial infarction, only single vessel involvement or no lesion greater than or equal to 50 to 69%. Tomography did not appear to provide improved diagnosis in women or in detection of disease in the right coronary artery. Although overall detection of coronary artery disease was not improved in patients with prior myocardial infarction, tomography provided improved identification of normal and abnormal vascular regions, particularly of the left anterior descending and circumflex artery regions. These results indicate that single photon emission computed tomography provides improved diagnostic performance compared with planar imaging in many clinical subgroups, and suggest that it represents the diagnostic imaging procedure of choice in exercise thallium-201 perfusion studies. PMID- 2563742 TI - The effect of azelastine on neutrophil and eosinophil generation of superoxide. AB - Azelastine is a new investigational drug used to treat rhinitis and asthma. In addition to described actions that include inhibition of immunologic release of histamine from mast cells and basophils, blockade of smooth muscle contraction to various spasmogenic mediators, and relaxation of airway smooth muscle, azelastine has been ascribed anti-inflammatory properties. To understand further the mechanisms by which azelastine may regulate inflammation, its effect on neutrophil and eosinophil superoxide (O2-) generation was evaluated. Purified suspension of neutrophils (greater than 95% pure) and eosinophils (greater than 85% pure) were isolated from human peripheral blood samples by continuous density gradients of Percoll. The isolated granulocyte suspensions (1 x 10(5) cells) were added to 96-well microtiter plates in the presence of cytochrome c, activated by either N-formyl-methionyl-leucyl-phenylalanine, phorbol myristate acetate, calcium ionophore A23187, or opsonized zymosan particles; O2- generation was measured by the reduction of cytochrome c. We found that azelastine significantly inhibited both neutrophil and eosinophil generation of O2- in a dose-dependent fashion (10(-7) to 10(-5) mol/L) with each activator except zymosan. Furthermore, the degree to which azelastine suppressed O2- was similar in neutrophils and eosinophils. Thus, azelastine, in concentrations achieved therapeutically, inhibited granulocyte generation of O2-. This anti-inflammatory effect may also be beneficial in the treatment of asthma. PMID- 2563743 TI - Evaluation of the efficacy and safety of loratadine in perennial allergic rhinitis. AB - Loratadine, a new nonsedating antihistamine, was evaluated for efficacy and safety in 228 patients with perennial allergic rhinitis. Taken at a dose of 10 mg once daily, loratadine was significantly more effective than placebo and comparable to terfenadine, 60 mg taken twice daily, in reducing combined symptom scores in this patient population. Efficacy was maintained throughout the 28-day course of treatment. The overall incidence of side effects with loratadine was low (14%) with few occurrences of sedation (3%) and dry mouth (4%). PMID- 2563744 TI - Duration of the effect of a single dose of azelastine on histamine-induced bronchoconstriction. AB - We studied the duration of the protective effect of azelastine against histamine induced bronchoconstriction in six subjects with asymptomatic asthma. The study was performed in two periods of five consecutive days each. After a histamine inhalation test, we randomly administered either placebo or a single oral dose of 8.8 mg azelastine in a double-blind crossover fashion. Histamine challenges were repeated 5 hours after ingestion and at 1 PM on the following four days. The geometric means of the dose of histamine (in cumulative breath units [cbu]) necessary to increase specific airway resistance by 100% as compared with baseline (PD100SRaw) were 8.7 and 8.5 cbu before placebo and azelastine, respectively. Placebo did not significantly influence PD100SRaw within 99 hours after treatment. Five and 27 hours after azelastine, PD100SRaw increased to 178.2 and 46.7 cbu, respectively (p less than 0.05). Two patients showed a highly significant protection against the airway effect of histamine even 99 hours after ingestion (p less than 0.01). These data demonstrate a variable duration of the antihistaminic property of azelastine. The prolonged therapeutic effect in some patients may be beneficial in the timing of medication intervals. PMID- 2563745 TI - Antihistamines in asthma. AB - Substantial evidence indicates that airway hyperresponsiveness in asthma is associated with the inflammatory response directed toward the airway epithelium and submucosa. Endogenously released spasmogenic mediators interacting with smooth muscle have a greater effect on hyperresponsive than on normal airways. Histamine, a bronchoactive and vasoactive mediator, is released in patients with naturally occurring or allergen-induced asthma. It is clear that mast cells are activated as part of the asthmatic response, and they have now been implicated as effector cells of mediator secretion based on the results of provocative tests. The advent of more potent histamine antagonists selective for H1 receptors has led to a reappraisal of antihistamine therapy in asthma. Drugs such as terfenadine and azelastine inhibit early bronchoconstriction and to some extent the late-phase asthmatic response as well. The efficacy of other new antihistamines (e.g., ketotifen and cetirizine) may extend beyond H1-receptor blockade to inhibit some of the chronic (and perhaps more important) cellular events in asthma. PMID- 2563746 TI - Systemic side effects of glaucoma medications. AB - A brief review of the common systemic side effects of the various medications used to treat glaucoma on an outpatient basis has been presented. This review is not complete, and the variety of side effects that may be caused by these drugs is extensive. Because it is usual for glaucoma patients to use several different drugs concurrently, the range of possible side effects in a given individual is large and serious. Not only should the ophthalmologist be familiar with the side effects of these medications, but all physicians involved in the patient's care should be aware of the glaucoma medications being used and consider these medications when the patient has symptoms or complications. Because the patient rarely associates the use of an eyedrop with problems in his or her general health, it is especially incumbent upon the physician to suspect and recognize these problems. Physicians should also elicit problems that the patient may not even complain of so alterations in therapy may be made. PMID- 2563747 TI - gamma-Glutamyltranspeptidase activity in intact leukocytes: flow cytometric analysis and sorting. AB - A fluorescent method developed for visualizing gamma-glutamyltranspeptidase (GGT) in intact liver cells was adapted to leukocytes and used in a multiparameter flow cytometric study of blood and bone marrow cells from rats with subcutaneous implants of mammary carcinoma 5A. The severe granulocytosis caused by this non metastatic tumor was preceded by a progressive rise in the percentage of leukocytes with high GGT fluorescence. Both granulocytes and small, immature cells of bone marrow showed increased GGT expression, whereas in blood this increase was attributable entirely to mature granulocytes. At 28 days (but not yet at 14 days) after carcinoma implantation, 20-30% of blood or bone marrow granulocytes constituted a distinct subpopulation in that their GGT fluorescence intensity range was much higher and did not overlap with the range for the rest of the population. The results indicate that fluorescent GGT assay of intact leukocytes provides a useful probe for flow cytometric analysis of population heterogeneity in leukoproliferative disorders. PMID- 2563748 TI - Serum electrolytes and parathyroid hormone in patients in a coronary care unit. AB - A prospective study was carried out in 499 patients admitted to a coronary care unit (CCU) in order to evaluate the incidence of clinically significant electrolyte disturbances. Low serum potassium values (less than 3.6 mmol l) occurred in 7% of the CCU patients and low serum magnesium values (less than 0.70 mmol l-1) in 6%. Few patients had low values of both these ions (1.9%). In 49 patients the contents of these electrolytes in muscle biopsies were similar to the values of control subjects and were unrelated to treatment with diuretics. Serum calcium was determined in 444 of the patients and was above the reference range in 11 (2.5%). If we consider their concomitant parathyroid hormone (PTH) values, primary hyperparathyroidism was likely to occur in at least seven patients (1.5%). Patients with acute myocardial infarction (AMI) had mean PTH and electrolyte values similar to those of individuals without this disease. In conclusion, the present study indicates that clinically important disturbances of magnesium, potassium or calcium homeostasis are rare among unselected patients admitted to a CCU. PMID- 2563749 TI - Individual identification from semen by the deoxyribonucleic acid (DNA) fingerprint technique. AB - For individual identification from semen, the deoxyribonucleic acid (DNA) fingerprint technique was used. In a blind trial, we succeeded in determining the semen donors among several volunteers comparing the DNA fingerprints of the blood and semen samples, respectively. Thereafter, we examined semen in a condom left beside a naked female dead body. The DNA fingerprint of the semen was recognized to be identical to that of the blood from a suspected man arrested later. This is the first report that the DNA fingerprint technique was practically used in a criminal investigation in Japan. PMID- 2563750 TI - 3H-spiperone binding sites in post-mortem brains from schizophrenic patients: relationship to neuroleptic drug treatment, abnormal movements, and positive symptoms. AB - In post-mortem putamen samples from 27 schizophrenics and 27 controls D2 receptors were measured by Scatchard analysis using 3H-spiperone as a ligand. Maximum number of binding sites (Bmax) and apparent dissociation constant (KD) were significantly increased only in patients in whom neuroleptic medication had been given within a three-month period before death. When the neuroleptic medication had been withdrawn at least 3 month before death, there was a slight, but not significant, reduction in Bmax values and unchanged KD values. Withdrawal of neuroleptic drugs was followed by a normalization of the KD values within 2 weeks and a slower reduction of Bmax values. There were 6 schizophrenic patients with mainly positive schizophrenic symptoms and 17 patients with mainly negative symptoms; positive schizophrenic symptoms were not related to higher Bmax values. There was no difference in 3H-spiperone binding between patients with and without movement disorders (tardive dyskinesia or extrapyramidal symptoms). PMID- 2563751 TI - Endocrine and neurochemical effects of (+)-PHNO, a dopamine D2 agonist. AB - The naphthoxazine compound, (+)-PHNO, is a dopamine D2 receptor agonist which acts within the central nervous system. The effects of this drug on serum concentrations of corticosterone and prolactin and on brain concentrations of catecholamines and some of their metabolites were determined in male rats. Administration of (+)-PHNO in doses ranging from 3-300 micrograms/kg i.p. resulted in increased serum corticosterone, decreased serum prolactin and decreased concentrations of the dopamine metabolites, DOPAC and HVA, in the brain. At the higher doses of (+)-PHNO, concentrations of MHPG sulfate in the brain stem were increased and hypothalamic epinephrine concentrations were decreased. Pretreatment with centrally acting dopamine antagonists (spiperone or haloperidol) prevented the (+)-PHNO-induced changes in serum corticosterone, prolactin and brain catecholamines. In contrast, pretreatment with halopemide, a dopamine antagonist which penetrates poorly into the brain, was unable to block the effects of (+)-PHNO on serum corticosterone and brain catecholamines. These data show that (+)-PHNO, a dopamine agonist structurally unrelated to other dopamine agonists, acts centrally to affect serum corticosterone and brain catecholamines. PMID- 2563752 TI - The influence of human corticotropin-releasing hormone on somatostatin secretion in depressed patients and controls. AB - Twenty subjects (10 patients with a major depressive episode and 10 individually matched healthy controls) received 100 micrograms synthetic human corticotropin releasing hormone (hCRH) as an i.v. bolus dose. Healthy subjects and depressed patients exhibited a significant increase of plasma somatostatin (SRIH) concentrations with no difference between both comparison groups. Compared to controls, depressed patients showed a significant attenuation of corticotropin (ACTH) responses, while cortisol secretion in response to hCRH was normal. No correlations were found among basal plasma concentrations of SRIH, ACTH or cortisol and SRIH, ACTH or cortisol responses following hCRH. These findings are compatible with the hypothesis that hypothalamic-pituitary-adrenal (HPA) hyperactivity in the depressive state may primarily be due to central hypersecretion of CRH and support the view of a hCRH-induced SRIH secretion which is not related to HPA dysfunction associated with major depression. PMID- 2563753 TI - Subcellular distribution of tyrosine hydroxylase in some catecholaminergic rat brain areas determined by a quantitative immunoblot assay. AB - The subcellular distribution of the protein tyrosine hydroxylase (TH) after fractionation of rat brain tissue was studied by a sensitive technique of immunoblot quantification in the dopaminergic nigrostriatal and the dorsal noradrenergic pathways and in the ventrolateral medulla. This repartition indicates that in all catecholaminergic regions of the cell bodies studied, the contribution of the nerve endings to the total TH amount is very low (less than 7%), in contrast to that observed in the terminal fields. The correlative subcellular determination of the TH amount and activity in the same tissue could be a useful approach for studying experimentally induced mechanisms of catecholamine synthesis modulation in different brain catecholaminergic pathways. PMID- 2563754 TI - Inositol phospholipid hydrolysis and potentiation of cyclic AMP formation by noradrenaline in rat cerebral cortex slices are not mediated by the same alpha adrenoceptor subtypes. AB - A pharmacological study was undertaken to determine whether the noradrenaline stimulated breakdown of inositol phospholipids and the potentiation of isoprenaline-stimulated cyclic AMP by noradrenaline in rat cerebral cortex slices are mediated by the same alpha-receptor subtype. The rank order of potency of a range of alpha 1 and alpha 2 antagonists suggests that both responses may involve an alpha 1 receptor, but there were several differences between the pharmacological profiles for the two systems. Although in both cases, all selective alpha 1 antagonists were more potent than alpha 2 antagonists, the rank orders and the absolute potencies differed for the two responses. The inhibition of the inositol phosphate response was characterised by a high alpha 1/alpha 2 antagonist ratio, and in most cases, Hill slopes of inhibition were consistent with the involvement of a single receptor site. Inhibition of the cyclic AMP response had a much lower alpha 1/alpha 2 antagonist ratio and generally exhibited Hill slopes less than one. Evidence has been provided suggesting that adenosine is involved in the potentiation of cyclic AMP and that other, as yet unidentified, factors may also be involved. Even in the absence of an adenosine component, the results presented support the suggestion that the potentiation due to noradrenaline is mediated by a receptor whose identity does not easily fit with the currently accepted classification of alpha adrenoceptors. PMID- 2563755 TI - Use of calcium antagonism for the characterization of drug-evoked dopamine release from the brain of conscious rats determined by microdialysis. AB - Dopamine was determined by microdialysis of the striatum of conscious rats. We investigated whether the release of dopamine, induced by nine different pharmacological treatments, was sensitive to calcium antagonism. Calcium antagonism was determined by Mg2+ or Cd2+ infusion. The following conditions were investigated: haloperidol, haloperidol plus GBR 12909, nomifensine, (+) amphetamine (all administered intraperitoneally), KCl, 1-methyl-4-phenyl pyridinium ion (MPP+), glutamate, ouabain, and 120 mmol/L magnesium (all applied by infusion through the dialysis membrane). The results on calcium antagonism were combined with data on tetrodotoxin (TTX) sensitivity. With the combined data, three different types of dopamine release were characterized. First, action potential-dependent dopamine release was observed in animals treated with saline, haloperidol, haloperidol plus GBR 12909, nomifensine, and ouabain. Second, action potential-independent release was established in the case of (+)-amphetamine, glutamate, MPP+, and 120 mmol/L Mg2+. Finally, K+-induced dopamine release was classified as TTX independent and calcium dependent. It is concluded that brain dialysis is a powerful method for differentiating between different types of neurotransmitter release. PMID- 2563756 TI - Effects of acute hyperammonemia on cerebral amino acid metabolism and pHi in vivo, measured by 1H and 31P nuclear magnetic resonance. AB - The effects of an acute intravenous infusion of ammonium acetate on rat cerebral glutamate and glutamine concentrations, energy metabolism, and intracellular pH were measured in vivo with 1H and 31P nuclear magnetic resonance (NMR). The level of blood ammonia maintained by the infusion protocol used in this study (approximately 500 microM, arterial blood) did not cause significant changes in arterial PCO2, PO2, or pH. Cerebral glutamate levels fell to at least 80% of the preinfusion value, whereas glutamine concentrations increased 170% relative to the preinfusion controls. The fall in brain glutamate concentrations followed a time course similar to that of the rise of brain glutamine. There were no detectable changes in the content of phosphocreatine (PCr) or nucleoside triphosphates (NTP), within the brain regions contributing to the sensitive volume of the surface coil, during the ammonia infusion. Intracellular pH, estimated from the chemical shift of the inorganic phosphate resonance relative to the resonance of PCr in the 31P spectrum, was also unchanged during the period of hyperammonemia. 1H spectra, specifically edited to allow quantitation of the brain lactate content, indicated that lactate rose steadily during the ammonia infusion. Detectable increases in brain lactate levels were observed approximately 10 min after the start of the ammonia infusion and by 50 min of infusion had more than doubled. Spectra acquired from rats that received a control infusion of sodium acetate were not different from the spectra acquired prior to the infusion of either ammonium or sodium acetate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563757 TI - L-[3H]glutamate binding to a membrane preparation from crayfish muscle. AB - The binding of L-[3H]glutamate to an isolated membrane preparation from crayfish tail muscle has been studied. The muscle homogenate was osmotically shocked, frozen and thawed, and thoroughly washed before incubation with L-[3H]glutamate. The preparation showed high specific binding of L-glutamate with a KD of 0.12 microM and Bmax of 4.7 pmol/mg protein measured in Tris/HCl pH 7.3 and at 4 degrees C. Nonspecific binding was 5-10% of total binding. The glutamate binding was highly stereospecific [K0.5 (D-glutamate), 270 microM] and showed a high degree of discrimination between L-glutamate and L-aspartate [K0.5 (L-aspartate), 54 microM]. In mammalian CNS preparations potent agonists of L-glutamate such as kainate and N-methyl-D-aspartate had no effect at 1 mM, and quisqualate was a weak inhibitor of L-glutamate binding [K0.5 (quisqualate), 162 microM]. Ibotenate was the most potent inhibitor [K0.5 (ibotenate), 0.27 microM], and various esters of L-glutamate were of intermediate potency as displacers of L-[3H]glutamate binding (K0.5 values from 6 to 60 microM). The glutamate binding site from crayfish muscle is clearly different from any of the subclasses of glutamate receptors in mammalian CNS. A possible physiological function of the binding site is a postsynaptic receptor for glutamate, either an extra-junctional or a junctional receptor. PMID- 2563758 TI - Regulation of thyrotropin releasing hormone degrading enzymes in rat brain and pituitary by L-3,5,3'-triiodothyronine. AB - The effect of treatment with L-3,5,3'-triiodothyronine (T3) on the levels of pyroglutamyl peptidase I and pyroglutamyl peptidase II in rat brain regions, pituitary, and serum was studied. Pyroglutamyl peptidase I cleaves pyroglutamyl peptides such as thyrotropin releasing hormone (TRH), luteinizing hormone releasing hormone, neurotensin, and bombesin, whereas pyroglutamyl peptidase II appears to be specific for TRH. Acute administration of T3 did not affect pyroglutamyl peptidase I in any of the regions studied, whereas pyroglutamyl peptidase II was significantly elevated in frontal cortex and pituitary. Treatment with T3 for 10 or 14 days significantly elevated pyroglutamyl peptidase I in pituitary, hypothalamus, olfactory bulb, hippocampus, and thalamus. Chronic T3 treatment elevated pyroglutamyl peptidase II in frontal cortex and in serum. These studies demonstrate regulation of neuropeptide degrading enzymes by thyroid hormones in vivo. This regulation may play a role in the negative feedback control of thyroid status by T3. PMID- 2563759 TI - Modulation of dendritic release of dopamine by N-methyl-D-aspartate receptors in rat substantia nigra. AB - A superfusion system was used to study the effects of excitatory amino acids (EAA) on release of [3H]dopamine ([3H]DA) previously taken up by rat substantia nigra (SN) slices. The EAA tested (20-250 microM), with the exception of quisqualate and kainate, markedly evoked [3H]DA release from nigral slices when Mg2+ ions were omitted from the superfusion medium. The EAA receptor agonists exhibited the following relative potency in stimulating [3H]DA release: L glutamate (L-Glu) greater than N-methyl-D-aspartate (NMDA) greater than NM(D,L)A greater than D-Glu much greater than quisqualate = kainate. D-2-Amino-5 phosphonovalerate (100-200 microM), an antagonist for NMDA receptors, substantially reduced [3H]DA release evoked by L-Glu or NMDA. In contrast, L-Glu diethyl ester (100-200 microM) produced a lesser blocking effect on [3H]DA release evoked by the EAA. Further experiments showed that the NMDA-mediated release of [3H]DA was totally suppressed by the omission of Ca2+ or by the addition of tetrodotoxin (0.1 microM) to the superfusion medium. In addition, strychnine, an antagonist for glycine (Gly) receptors, significantly decreased NMDA (100 microM)-evoked as well as glycine (100 microM)-evoked release of [3H]DA from nigral slices. The results shown support the idea that activation of NMDA subtype receptors in SN may trigger a Ca2+-dependent release of DA from dendrites of nigro-striatal DA-containing neurons. Furthermore, a transsynaptic mechanism that may partially involve Gly-containing interneurons is proposed to account for some of the events mediating NMDA receptor activation and DA release in SN. PMID- 2563760 TI - Pharmacological and functional heterogeneity of astrocytes: regional differences in phospholipase C stimulation by neuromediators. AB - Carbamylcholine stimulated phospholipase C activity in astrocytes in primary culture from the mesencephalon but not from the striatum or cerebral cortex of the mouse embryo. An alpha 1-adrenergic-mediated response was observed in all astrocyte populations. 2-Chloroadenosine potentiated the alpha 1-adrenergic response in mesencephalic and striatal astrocytes but not in cortical astrocytes. It also stimulated the carbamylcholine-evoked response in mesencephalic astrocytes. Through cell-cell cooperation, 2-chloroadenosine potentiated the neuronal carbamylcholine-evoked activation of phospholipase C in homotopic cocultures (neuro-glial) from the striatum but not in homotopic cocultures (neuro glial) from the cerebral cortex or in heterotopic cocultures (cortical astrocytes striatal neurons; striatal astrocytes-cortical neurons. PMID- 2563761 TI - A voltage-clamp study of isolated stingray horizontal cell non-NMDA excitatory amino acid receptors. AB - 1. Horizontal cells enzymatically isolated from retinas of the Atlantic stingray (Dasyatis sabina) were voltage-clamped using the patch electrode in the whole cell mode. A rapid microsuperfusion system was used to apply excitatory amino acid agonists and antagonists. 2. The isolated cells responded to glutamate (GLU), kainate (KA), quisqualate (QA) and alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA). Responses elicited by GLU, QA, and AMPA but not KA exhibited a concentration-dependent and concanavalin A- (Con-A) sensitive desensitization. No responses were elicited by aspartate, N-methyl-D-aspartate, or quinolinate at concentrations as high as 1.0 mM. 3. Judging from the concentration producing one-half of the maximal current response (EC50), the rank order affinities of the agonists was QA greater than or equal to GLU greater than AMPA greater than KA. Whereas KA had the lowest affinity of the agonists tested it was the most efficacious, producing the largest currents. Hill coefficients of the concentration-response data were near two for KA and GLU and near one for QA and AMPA. 4. The agonists differed in their sensitivity to various excitatory amino acid receptor antagonists. Kynurenate (KYN) produced a nearly complete block of horizontal cell responses to GLU and KA at concentrations that had little effect on QA and AMPA. Piperidine-2,3-dicarboxylic acid (cis-PDA), 1-(4 chlorobenzoyl)-piperazine-2,3-dicarboxylic acid (pCB-PzDA), and folic acid were less potent antagonists than KYN but were also better blockers of KA and GLU responses than of QA- and AMPA-elicited responses. 5. When QA, AMPA, or GLU were applied in combination with 55.0 microM KA the current was less than that produced by KA alone. The rank order potency for the inhibition of KA-elicited responses was QA greater than AMPA greater than GLU. In the presence of low concentrations of KA (1.0-20.0 microM), QA- and AMPA-elicited responses were potentiated. This potentiation was prevented by KYN. 6. Single-channel conductance and mean open time were estimated from the current noise fluctuations in the presence of agonist. The mean single-channel conductance for QA was 9 pS that was almost twice as large as the conductance for KA (5.9 pS) and GLU (5.7 pS). The mean open time in the presence of QA or GLU was approximately 1 ms, which was about one-half of that for KA (2.0 ms). 7. These results are best explained by the existence of a single receptor protein with multiple but not identical ligand-binding sites.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2563762 TI - Selective modulation of spike duration by serotonin and the neuropeptides, FMRFamide, SCPB, buccalin and myomodulin in different classes of mechanoafferent neurons in the cerebral ganglion of Aplysia. AB - An examination of the cellular properties and synaptic outputs of mechanoafferent neurons found on the ventrocaudal surface of the cerebral ganglion of Aplysia indicated that the cerebral mechanoafferent (CM) neurons are a heterogeneous population of cells. Based on changes in action potential duration in response to bath applications of 5-HT in the presence of TEA, CM neurons could be divided into 2 broad classes: mechanoafferents whose spikes broaden in response to 5-HT (CM-SB neurons) and mechanoafferents whose spikes narrow in response to 5-HT (CM SN neurons). Morphological and electrophysiological studies of the CM-SN neurons indicated that they were comprised of previously identified interganglionic cerebral-buccal mechanoafferent (ICBM) neurons and a novel set of sensory neurons that send an axon into the LLAB cerebral nerve and have perioral zone receptive fields that are similar to those of ICBM neurons. Changes in spike width due to 5 HT were correlated with changes in synaptic output as indicated by the magnitudes of EPSPs evoked in postsynaptic neurons. Electrical stimulation of cerebral nerves and connectives also produced spike narrowing or broadening, and the sign of the effect was a function of the parameters of stimulation. Both heterosynaptic facilitation and heterosynaptic depression of EPSPs evoked in follower cells could be demonstrated. A variety of putative neuromodulators other than 5-HT were also found to affect the duration of action potentials in both classes of CM neurons. FMRFamide had effects opposite to that of 5-HT. SCPB and a recently characterized Aplysia neuropeptide, buccalin, broadened the spikes of both CM classes. Another neuropeptide, myomodulin, decreased the duration of CM SB neuron spikes but had no effect on CM-SN spikes. Since the CM neurons appear to mediate a variety of competing behaviors, including feeding, locomotion, and defensive withdrawal, the various neuromodulator actions may contribute to the mechanisms whereby behaviors are selected and modified. PMID- 2563763 TI - Signaling properties of Ascaris motorneurons: graded active responses, graded synaptic transmission, and tonic transmitter release. AB - The commissural motorneurons of the nematode Ascaris are capable of transmitting signals passively over long distances with little decrement. This ability is due to the high resistivities of their membranes (Davis and Stretton, 1989). Although these cells rely on their passive properties for long-distance signaling, voltage sensitive channels are present in commissural membranes. These channels underlie the graded active responses that can be elicited at the offset of abrupt hyperpolarizing and depolarizing intracellular current pulses. The inhibitory motorneurons generate membrane potential oscillations when they are strongly depolarized. All-or-none action potentials have never been observed to occur spontaneously, nor has it been possible to evoke them even when the cells have been strongly hyperpolarized to remove any possible channel inactivation. Our findings indicate that the typical all-or-none action potentials so commonly used in nerve cells throughout the animal kingdom do not occur in these cells. Synaptic transmission is therefore mediated without spikes and is graded. The resting potentials of Ascaris motorneurons lie where the synaptic input-output curves are steepest, above the threshold for release of neurotransmitter. Tonic transmitter release from commissural motorneurons may be the neural mechanism underlying the hydrostatic skeleton of Ascaris. PMID- 2563764 TI - Postnatal development of glutamatergic, GABAergic, and cholinergic neurotransmitter phenotypes in the visual cortex, lateral geniculate nucleus, pulvinar, and superior colliculus in cats. AB - We have analyzed the postnatal development of glutamatergic/aspartergic, GABAergic, and cholinergic neurotransmitter systems in the visual cortical Areas 17 and 18, lateral geniculate nucleus (LGN), pulvinar, and the visual and non visual parts of superior colliculus (SC) in kittens. High-affinity uptake of D aspartate (HA D-Asp), glutamate decarboxylase (GAD), and choline acetyltransferase (ChAT) activities were measured as a means of probing the development of the respective transmitter systems. HA D-Asp exceeded the adult level several-fold in all areas during the postnatal period which corresponded with the period of maximal dendritic/axonal branching patterns and synapse densities in the respective regions. GAD exhibited a gradual increase towards adult levels during the first month. The adult level was reached during postnatal week (PNW) 5-6 in Areas 17 and 18, during PNW3 within LGN, pulvinar, and the visual part of SC. In the nonvisual part of SC, the adult GAD level was reached as early as PNW2. ChAT exhibited biphasic developmental profiles in Areas 17 and 18. An initial peak of near adultlike activity in PNW2 was followed by a decline and subsequently by a slow increase towards adult levels during PNW5-17. ChAT developed very slowly in LGN and pulvinar, and in the latter structure only approximately 70% of the adult activity had been attained by PNW17. In both subdivisions of SC, ChAT had reached adult levels during PNW3-5. Dark-rearing from birth until PNW6 moderately attenuated GAD development in all areas and increased ChAT activity in Areas 17 and 18 but did not affect development of HA D Asp in any part of the kitten visual system. Our neurochemical findings in the developing cat visual system are consistent with available evidence regarding localization of neurotransmitter systems, as well as postnatal changes in terms of cytoarchitectonics, synaptogenesis, functional development, and susceptibility to neonatal dark-rearing in visual pathways. PMID- 2563765 TI - Human fetal mesencephalic tissue grafted to dopamine-denervated striatum of athymic rats: light- and electron-microscopical histochemistry and in vivo chronoamperometric studies. AB - Human fetal mesencephalic tissue obtained from elective first-trimester abortions was grafted to 6-hydroxydopamine-denervated striatum of athymic (nude) rats. After 3-6 months, the transplants were evaluated by light and electron microscopy using antibodies against tryosine hydroxylase (TH), human specific Thy-1 (Thy-1), 5-hydroxytryptamine (5-HT), and laminin. In vivo chronoamperometric studies of K+ induced release of electroactive species were done prior to the histochemical evaluations. At the light microscopical level, Thy-1-immunoreactivity was evenly distributed throughout the entire transplants. Thy-1-immunoreactive nerve fibers were observed radiating from the graft into the host striatum. In sections that were double-stained with antibodies against Thy-1 and TH, such nerve fibers contained both markers. Also 5-HT-immunoreactive cells were found in the grafts with processes both in the grafts and radiating into host neuropil. Laminin immunohistochemistry showed an even distribution of capillaries in the graft with less density than in host brain, suggesting immaturity of graft tissue. At the ultrastructural level, TH-immunoreactive axons made symmetric contacts with unlabeled dendritic shafts and dendritic spines within the host brain. A few asymmetric contacts with TH-immunoreactive axons were seen. 5-HT-immunoreactive terminals made both symmetric and asymmetric contacts with unlabeled dendritic shafts and spines. In vivo chronoamperometry using local application of K+ revealed average signals that were lower on the transplanted side than in control striatum. However, close to the grafts significant amounts of the K+-evoked signal amplitudes were as large as 1.3 microM, and the ratio of the reduction to oxidation currents suggested release of a mixture of dopamine and 5-HT. Taken together, this study shows that human fetal mesencephalic tissue pieces survive grafting into nude rats, develop normal vascularization, and express coexistence of TH- and Thy-1-immunoreactivity. Human TH- and 5-HT-immunoreactive nerve fibers form synapses in host striatum and release monoamine neurotransmitters. PMID- 2563766 TI - Stereotypic morphology of glutamatergic synapses on identified muscle cells of Drosophila larvae. AB - The distribution and morphology of glutamatergic synapses on Drosophila bodywall muscle fibers were examined at the single-synapse level using immunocytochemistry and electrophysiology. We find that glutamate-immunoreactive motor endings innervate the entire larval bodywall musculature, with each muscle fiber receiving at least one glutamatergic ending. The innervation is initiated at stereotyped locations on each muscle fiber from where moderately branched varicose nerve processes project over the internally facing muscle surface. Individual muscle fibers have distinct stereotypic patterns of nerve endings that occupy characteristic regions on the cell surface. The muscle-specific branching pattern of motor endings is reiterated by segmentally homologous fibers. Two morphological types of innervating nerve processes can be distinguished by their bouton size distributions: (1) Type I processes, which have localized branching and a broad size distribution of relatively large varicosities ranging up to 8 microns (mean diameter, 3.1 +/- 1.6 microns; +/- SD, n = 521), and (2) thinner Type II processes, which have a narrower distribution of small varicosities with a mean diameter of only 1.4 +/- 0.6 microns (+/- SD, n = 214). Immunoelectron microscopy with peroxidase-labeled second antibody demonstrates that the varicosities are surrounded by a subsynaptic reticulum, that they contain immunoreactive vesicles of about 30-50 nm, and thus probably represent synaptic release sites. By iontophoretic application of glutamate we mapped the responsive sites on the muscle surface and found an excellent correspondence between transmitter sensitivity and the patterns of endings as described by immunocytochemistry. In contrast to our finding of numerous glutamate iontophoresis-sensitive sites, we did not detect any aspartate-responsive muscles. These data provide strong new evidence for glutamate being an endogenous transmitter at the Drosophila larval neuromuscular junction. PMID- 2563767 TI - Brugia malayi and Brugia pahangi: inherent difference in immune activation in the mosquitoes Armigeres subalbatus and Aedes aegypti. AB - The inherent ability of Brugia malayi and Brugia pahangi (Nematoda) to establish successful relationships with the mosquitoes Armigeres subalbatus and Aedes aegypti Liverpool strain was evaluated. Brugia pahangi microfilariae (mff) avoided the immune response and developed normally in A. subalbatus exposed to the parasite by an infective bloodmeal, whereas nearly 85% of B. malayi were destroyed by the immune response. Because A. aegypti supports the development of both filarial worm species but destroys intrathoracically inoculated B. pahangi isolated from jird blood, blood-isolated B. malayi were inoculated into A. aegypti, and the immune response was compared with that observed against B. pahangi. The response against B. malayi was significantly more rapid and effective than the response against B. pahangi. Similar results were obtained when blood-isolated B. pahangi or B. malayi were inoculated into A. subalbatus. Microfilariae of both species were able to avoid immune destruction in A. aegypti if they were allowed to penetrate the Liverpool midgut in vitro prior to inoculation. Most B. pahangi that had first penetrated an Armigeres midgut prior to inoculation into A. subalbatus were able to avoid the immune response, but by day 3 postinoculation, less than 40% of the B. malayi, treated in the same manner, were able to escape the immune response. Genetic susceptibility of mosquitoes to infection by filarial worms and potential mechanisms of immune evasion/suppression are discussed regarding B. malayi and B. pahangi. PMID- 2563768 TI - Mechanisms of central hemodynamic and sympathetic regulation by mu opioid receptors: effects of dermorphin in the conscious rat. AB - The effects of i.c.v. administered dermorphin, a highly selective mu-opioid agonist, on cardiac function and renal, mesenteric and hindquarter blood flow were studied in conscious rats. Core temperature, blood gases, arterial plasma levels of norepinephrine, epinephrine, dopamine, 3,4-dihydroxyphenylalanine and dihydroxyphenylacetic acid (DOPAC) also were examined. Cardiac output was measured using a thermodilution technique and regional blood flows using directional pulsed Doppler velocimetry. Dermorphin, at doses of 0.1-100 nmol/kg, increased blood pressure and hindquarter blood flow, renal and mesenteric resistance, and core temperature. Higher doses (1-5 mumol/kg) caused respiratory depression, acidosis, and shock despite profound sympatho-adrenomedullary stimulation. Circulating levels of catecholamines were significantly increased at the dermorphin doses of 0.1-100 nmol/kg. At the 100 nmol/kg dose, plasma levels of epinephrine, norepinephrine, the dopamine metabolite dihydroxyphenylacetic acid and the catecholamine precursor 3,4-dihydroxyphenylalanine were increased by 2-15-fold. The data indicate that mu opioid receptor stimulation exerts potent effects on cardiorespiratory functions, activates the sympathoadrenomedullary system and produces a pattern of blood flow changes consistent with the stress induced "defense" response (skeletal muscle vasodilation and splanchnic vasoconstriction). Excessive mu opioid receptor stimulation leads to shock due to respiratory and hemodynamic collapse. PMID- 2563769 TI - Characterization of L-glutamate action on the release of endogenous dopamine from the rat caudate-putamen. AB - In the present study the effect of L-glutamic acid (L-Glu), N-methyl-D-aspartic acid (NMDA), kainic acid (KA) and quisqualic acid (QUIS) on the release of endogenous dopamine (DA) from slices of the rat caudate-putamen was investigated. DA was measured by high-performance liquid chromatography coupled to an electrochemical detector. L-Glu, NMDA, KA and QUIS, in the absence of Mg++, produced a dose-related, Ca++-dependent increase in DA release. The order of agonist efficacy was L-Glu greater than NMDA greater than KA = QUIS. D-2-amino-7 phosphonoheptanoic acid (0.5 mM), but not L-2-amino-7-phosphonoheptanoic acid, antagonized the action of L-Glu and NMDA, but did not modify the effect of KA or QUIS. Tetrodotoxin (0.1 microM) partially inhibited the stimulatory effect of KA and QUIS, but not that of L-Glu or NMDA. Mg++ (1.2 mM) abolished the excitatory effect of NMDA, significantly reduced the action of L-Glu, but did not influence the action of KA or QUIS. The inhibitory action of Mg++ on the L-Glu-induced DA release was reversed when L-Glu was coupled to high concentrations of K+. N allylnormetazocine (SKF-10,047), a benzmorphan agent, produced a stereospecific inhibition of L-Glu-induced DA release. This inhibition was also produced by 1-[1 (2-thienyl)cyclohexyl]piperidine, a phencyclidine receptor ligand, but not by 1,3 di-O-tolylguanidine, a sigma receptor-selective ligand. The results of this study show that L-Glu increases DA release predominantly by activation of the NMDA receptor located presynaptically on dopaminergic afferents.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563770 TI - Tertatolol increases glomerular filtration and urinary sodium in anesthetized rats. AB - Tertatolol is a new beta-adrenergic blocking agent that possesses unique actions on kidney function. In the present study, the effect of increasing doses of tertatolol (0.025, 0.05, 0.1, 0.2 and 0.3 mg) was examined in different groups of anesthetized rats and the results compared with injection of the vehicle. Bolus i.v. injections of tertatolol resulted in a dose-related elevation of urine volume (UV), from 4.1 +/- 0.3 to 6.0 +/- 0.4, 10.2 +/- 1.3, 13.8 +/- 1.2, 18.8 +/ 1.9 and 16.4 +/- 1.8 microliters/min, respectively. Similarly, urine Na increased from 0.4 +/- 0.1 to 0.8 +/- 0.2, 1.2 +/- 0.2, 2.2 +/- 0.3, 3.4 +/- 0.7 and 2.9 +/- 0.6 mueq/min. These changes were associated with a progressive rise in glomerular filtration rate (GFR) from a mean control value of 1.15 +/- 0.10 to 1.28 +/- 0.09, 1.41 +/- 0.07, 1.50 +/- 0.10, 1.62 +/- 0.16 and 1.48 +/- 0.07 ml/min. No change in urinary phosphate excretion was observed. In a separate group of studies, the effect of tertatolol was compared with propranolol (1 mg): the maximal elevation on UV and urine Na was less than 25% of that obtained with tertatolol. Elevation of GFR was also 30% higher in animals receiving tertatolol. The effect of tertatolol on UV and urine Na resulted in part from the rise in GFR but also from tubular inhibition of Na transport, since fractional Na excretion increased from 0.24 +/- 0.09 to a maximal value of 1.76 +/- 0.10%. This tubular action presumably occurs beyond the proximal nephron, because no effect on urinary phosphate excretion was obtained during tertatolol.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563771 TI - Nitroglycerin-induced tolerance affects multiple sites in the organic nitrate bioconversion cascade. AB - The effect of nitroglycerin (NTG)-induced tolerance on the spasmolytic effects of a series of vasodilators was determined to establish potential sites of tolerance. Concentration-effect curves to vasodilators were completed concurrently in U46619-contracted bovine isolated coronary artery rings pre exposed to 100 microM NTG for 10 min (NTG-tolerant rings) and in control rings not pre-exposed to NTG. Compared to control rings, NTG-tolerant rings were markedly less responsive (P less than .01, n = 8-10) to the spasmolytic actions of NTG, isosorbide dinitrate, sodium nitroprusside (SNP) and 3 morpholinosydonimine (SIN-1), whereas the spasmolytic actions of S-nitroso-N acetylpenicillamine and nitric oxide were only marginally attenuated in NTG tolerant rings. On the other hand, no significant difference in the relaxant responses of NTG-tolerant and control coronary artery rings were observed to either the endothelium-dependent vasodilator, A23187 or the guanylate cyclase independent vasodilator, theophylline. In additional cross-tolerance studies, relaxations to the organic nitrate vasodilator, NTG were significantly more attenuated (P less than .05, n = 5) by tolerance induced by NTG, than by either SNP or SIN-1, whereas the actions of the non-nitrate vasodilators, SNP and SIN-1 were attenuated more by SNP-and SIN-1-induced tolerance than by NTG-induced tolerance (P less than .05, n = 5). We conclude that, in this isolated coronary artery preparation, NTG-induced tolerance affects at least two major sites in the cascade of events between the initial site of NTG action and guanylate cyclase activation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563772 TI - 3-aminopyridazine derivatives with atypical antidepressant, serotonergic, and dopaminergic activities. AB - Minaprine [3-[(beta-morpholinoethyl)amino]-4-methyl-6-phenylpyridazine dihydrochloride] is active in most animal models of depression and exhibits in vivo a dual dopaminomimetic and serotoninomimetic activity profile. In an attempt to dissociate these two effects and to characterize the responsible structural requirements, a series of 47 diversely substituted analogues of minaprine were synthesized and tested for their potential antidepressant, serotonergic, and dopaminergic activities. The structure-activity relationships show that dopaminergic and serotonergic activities can be dissociated. Serotonergic activity appears to be correlated mainly with the substituent in the 4-position of the pyridazine ring whereas the dopaminergic activity appears to be dependent on the presence, or in the formation, of a para-hydroxylated aryl ring in the 6 position of the pyridazine ring. PMID- 2563773 TI - New 4-(heteroanilido)piperidines, structurally related to the pure opioid agonist fentanyl, with agonist and/or antagonist properties. AB - A research program based on certain heterocyclic modifications (12-50) of the fentanyl (1) molecule has generated a novel class of opioids. In the mouse hot plate test, these compounds were weaker analgesics than 1. Two types of antagonists were observed in morphine-treated rabbits: those (e.g., 28) that reversed both respiratory depression and analgesia and those (e.g. 32) that selectively reversed respiratory depression. Evaluation of in vitro binding affinities to rat brain opioid receptors was inconclusive for a common locus of action for the agonist as well as the antagonist component. Further pharmacological evaluation of 32, N-(2-pyrazinyl)-N-(1-phenethyl-4-piperidinyl)-2 furamide, in the rat showed it to be a potent analgesic (ED50 = 0.07 mg/kg, tail flick test) with little cardiovascular and respiratory depression when compared to fentanyl. PMID- 2563774 TI - New mutation to Huntington's disease. AB - We report a large family with an isolated case of Huntington's disease (HD), which is probably the result of a new mutation. The patient developed clinical signs typical of HD at the age of 36. The clinical course of the patient's disease is documented by several clinical admissions over a period of 14 years at present. The family history is strikingly negative with the parents having been clearly unaffected into their 80s and with 13 older and two younger, living, healthy sibs. Extensive testing of polymorphic markers (blood groups, red cell and serum proteins, HLA antigens) showed no indication of non-paternity, but rather gave strong support to the hypothesis that the proband is a full sib. In addition, DNA typing for several RFLPs known to be closely linked to the HD gene locus indicated that several clearly unaffected sibs share one or the other or both of the patient's haplotypes. This is further evidence in favour of the hypothesis of a new mutation at the HD locus. The posterior probability of a new mutation to HD in the patient exceeds 99%, even if an a priori probability of non paternity of 10% and a mutation rate of HD of 10(-7) is assumed. PMID- 2563775 TI - Alpha thalassaemia: a potential source of error in DNA linkage studies for adult polycystic kidney disease. AB - DNA polymorphisms associated with adult polycystic kidney disease and detected by restriction enzymes PvuII and RsaI were sought in 18 members of a Greek Cypriot family. The adult polycystic kidney disease (APKD) defect was shown to segregate with a specific 3'HVR PvuII DNA polymorphism and enabled exclusion of APKD in two children incorrectly diagnosed by ultrasound. An unusual problem of the DNA linkage approach in detection of APKD is illustrated. This occurred after coinheritance of alpha+ thalassaemia and APKD, which resulted in an altered band size for the (+) RsaI restriction fragment length polymorphism. PMID- 2563776 TI - Carrier detection and early diagnosis of Wilson's disease by restriction fragment length polymorphism analysis. AB - Wilson's disease, a rare autosomal recessive disorder, has been recently mapped to the long arm of chromosome 13 (q14.1). In this study, we carried out linkage analysis between three chromosome 13 DNA markers, D13S1, D13S10, D13S2, the locus for the red cell enzyme esterase D (ESD), and the Wilson's disease locus (WND) in 17 Wilson's disease families of Italian descent, mostly from Sardinia. We confirmed a tight linkage [theta = 0.00, Z (theta) = 4.07] between the WND and ESD loci, and provided suggestive evidence for linkage [theta = 0.00, Z(theta) = 1.85] of the WND locus with D13S10. Multipoint linkage analysis indicated the following order: centromere-D13S1-D13S10-WND-ESD-D13S2. RFLP analysis at these two loci in our families allowed us either to define the carrier status (50%) or to exclude the homozygous state (25%) in the great majority of unaffected sibs. PMID- 2563777 TI - Prenatal exclusion testing for Huntington's disease: a problem of too much information. AB - At eight weeks of pregnancy a couple were informed that the prospective father's mother had died of Huntington's disease (HD). There were no living affected members in the immediate family to confirm the diagnosis. By inspection of the local genetic register, it was established that it was indeed HD segregating in the extended family. Genotyping of the prospective mother and father, the father's unaffected father, and his unaffected maternal grandmother was carried out using a battery of polymorphic DNA markers, including a new probe which has a very low recombination rate with the HD locus. Analysis of DNA from a chorionic villus sample taken at 10 weeks of pregnancy showed that the fetus must have inherited a chromosome from its father's affected mother. Its risk of developing HD was 47%. If the genotype of the unaffected maternal grandmother was taken into account, the risk was reduced to 42%. Neither risk was considered acceptable by the prospective parents and the pregnancy was terminated at 12 weeks' gestation. Prospects for future pregnancies are good, with a 50% chance of having a child whose risk of inheriting the HD gene is less than 1.5%. In retrospect it was noted that although genotyping of the maternal grandmother had refined the fetal risk, it had also nearly contributed to an inadvertent and unwanted predictive test for HD on the father. This case makes the point that in prenatal exclusion testing, linkage information must be generated with considerable care. PMID- 2563778 TI - The cardiocirculatory and metabolic effects of endotoxin challenge after canine resuscitated hemorrhagic shock. AB - Although adequate volume resuscitation has decreased mortality from hemorrhagic shock, recovery in many patients is complicated by sepsis. To determine whether a subject debilitated by hemorrhagic shock would exhibit greater cardiocirculatory dysfunction when challenged with sepsis, ten dogs (Group I) were hemorrhaged to a mean arterial blood pressure of 30 mm Hg. After 2 hours of hypotension, shed blood and lactated Ringer's solution (50 ml/kg) were given, and the dogs were observed for 3 to 6 days. Ten dogs were sham hemorrhage and served as controls (Group II). On the experimental day, all cardiovascular and hemodynamic parameters were measured in both groups of animals before endotoxin challenge. There was no significant difference in cardiac output, stroke volume, stroke work, +dP/dt max, myocardial blood flow, myocardial oxygen metabolism, or acid base balance in the two groups. Compared to sham-hemorrhaged dogs, resuscitated shock dogs had a significantly lower mean arterial blood pressure (127 +/- 7 vs. 110 +/- 6 mm Hg; p less than 0.05), and heart rate was significantly higher (86 +/- 6 vs. 109 +/- 7 beats/minute; p less than 0.05). Furthermore, maximal rate of left ventricular pressure fall (-dP/dT max) was significantly lower in the animals previously hemorrhaged, suggesting a persistent defect in left ventricular relaxation. Blood glucose and insulin levels were significantly elevated in the resuscitated shocked dogs, likely due to increased circulating catecholamine concentrations and enhanced glycogenolysis. Endotoxin shock caused significant hypotension, acidosis, and impaired regional perfusion in all dogs. In addition, cardiac output, stroke volume, dP/dT, and left ventricular end diastolic pressure fell and hyperglycemia and hyperinsulinemia occurred in all dogs after endotoxin injection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563780 TI - Bilateral ureteral stricture from polyarteritis nodosa. AB - A case of bilateral, asynchronous ureteral stricture from polyarteritis nodosa is described. Two cases of unilateral ureteral stricture from polyarteritis nodosa have been reported previously. Ureteral obstruction not associated with retroperitoneal fibrosis is rare with polyarteritis nodosa. PMID- 2563779 TI - Detection of missing vesicoureteral reflux with bethanechol chloride-aided voiding cystourethrography. AB - Experience in the detection of absent vesicoureteral reflux with bethanechol aided voiding cystourethrography is presented. The detection rate with conventional contrast-enhanced voiding cystourethrography among 9 study subjects, in whom vesicoureteral reflux otherwise was highly suspected from accompanying clinical, urographic and endoscopic features, and in whom it was absent contralaterally or bilaterally, was improved from 33 per cent (only 3 of 9 patients were positive for reflux) to 100 per cent (all 9 were proved to have vesicoureteral reflux) after bethanechol stimulation. Some representative cases are presented and the mechanisms of inducing vesicoureteral reflux with bethanechol are discussed as well as its clinical relevance. Bethanechol-aided voiding cystourethrography is suggested as a highly sensitive method to detect absent vesicoureteral reflux. PMID- 2563781 TI - Can the durability of mitral valve reconstruction be increased? PMID- 2563782 TI - Rat hippocampal slices 'in vitro' display spontaneous epileptiform activity following long-term organotypic culture. AB - Organotypic cultures of rat hippocampal slices were maintained for periods of up to 12 weeks in vitro. Cultures adopted a two-dimensional architecture whilst retaining the subfields characteristic of intact hippocampal slices. Coventional intracellular onset of spontaneous long-lasting epileptiform activity. Epileptiform activity characteristic of both interictal and ictal events (paroxysmal depolarising shifts, tonic/clonic phases and afterdischarges) was observed in the absence of pharmacological manipulation or of orthodromic stimulation. Epileptiform activity was abolished in the presence of high Mg2+ concentration or tetrodotoxin, agents known to block synaptic transmission. In addition, the frequency of epileptiform events was independent of membrane potential and the amplitude of the paroxysmal depolarising shift (PDS) displayed a near linear relationship with membrane potential. The PDS could be reversed at potentials approaching synaptic equilibrium potential. The N-methyl-D-aspartate (NMDA)-receptor antagonist DL-2-amino-5-phosphonovalerate (DL-APV) dose dependently reduced both the amplitude and duration of the spontaneous paroxysmal shift, having no effect on the initiation of the event or the resting membrane parameters of the neurone. DL-APV also attenuated a late component of the synaptically evoked excitatory postsynaptic potentials (epsp) not observed in non epileptiform neurones. Application of GABAA receptor antagonists bicuculline or picrotoxin converted interictal events to ictus. In the presence of these agents, ictal events were up to 90 s in duration. These results suggest that long-term culturing of hippocampal explants leads to an alteration in the balance of excitatory and inhibitory synaptic activity. This allows the expression of an excitatory amino acid depolarisation acting through NMDA receptors which contributes to the generation and maintenance of spontaneous epileptiform activity which is synaptic in origin. PMID- 2563783 TI - Beta-adrenergic blocking drugs in the treatment of congestive heart failure. AB - Experimental and clinical data suggests that increased serum catecholamines may have predominantly detrimental effects in patients with congestive heart failure. Some investigators have proposed use of beta-blockers in heart failure as a means of ameliorating the harmful effects of the excess catecholamines. The clinical experience to date with use of these agents as therapy in congestive heart failure is limited but does suggest a future role for beta-blockers as adjunct therapy in a select population of patients with heart failure. PMID- 2563784 TI - Beta-adrenergic blockade and calcium channel blockade in myocardial infarction. AB - Because of their hemodynamic and antiarrhythmic actions, beta-adrenergic blockers and calcium-entry blockers have been suggested for use in patients with myocardial infarction (MI) for reducing infarct size, preventing ventricular ectopy, and for prolonging life in survivors of acute MI. Experimental studies have suggested their usefulness in these areas. Clinical studies have demonstrated a role for beta-blockers in the hyperacute phase of MI, and in longterm treatment of infarct survivors. Calcium channel blockers appear to have somewhat less utility in patients with Q wave MIs, but may have an important role in therapy of the non-Q wave infarct. PMID- 2563785 TI - [The AIDS conference and reality]. PMID- 2563786 TI - Fall in blood pressure with modest reduction in dietary salt intake in mild hypertension. Australian National Health and Medical Research Council Dietary Salt Study Management Committee. AB - 111 untreated subjects (mean [SEM] age 58.4 [1.0] years; 93 male, 18 female) with diastolic blood pressure between 90 and 100 mm Hg were seen fortnightly, and after four pre-diet visits they were randomised into a low sodium intake group (53 subjects; diet containing less than 80 mmol sodium/day plus 8 placebo tablets daily) or a normal sodium intake group (55 subjects; same dietary sodium plus 8 slow-release sodium chloride [10 mmol] tablets daily). 103 subjects completed the intervention phase of 8 weeks. Urinary sodium fell significantly in the low sodium group but not in the normal sodium group. Urinary potassium excretion did not change in either group. Mean (SEM) systolic and diastolic blood pressure fell by 6.1 (1.1) and 3.7 (0.6) mm Hg, respectively, in the low sodium group, but by only 0.6 (1.0) and 0.9 (0.6) mm Hg in the normal sodium group. Multivariate analysis allowing for the effects of pre-diet blood pressure, weight, and age, reduced the effect of lowering the sodium intake on the systolic pressure from 5.5 (SEM 1.5) mm Hg to 4.8 (1.3) mm Hg (p less than 0.005) but the effect on diastolic pressure was not changed significantly. PMID- 2563787 TI - Noradrenergic activity and silent ischaemia in hypertensive patients with stable angina: effect of metoprolol. AB - 30 patients (10 normotensive, 20 hypertensive) with stable angina and positive treadmill exercise tests entered a double-blind, placebo-controlled crossover trial of metoprolol, 100 mg twice daily. At the end of each treatment phase, blood pressure was monitored for 24 h and Holter and real-time electrocardiographic (ECG) monitoring were carried out and an activity diary kept for 48 h. Blood samples for catecholamine measurement were taken after 30 min supine, 60 min standing, and at the first silent ischaemic event, triggered by the real-time ECG monitor, by means of an ambulatory blood withdrawal pump. Metoprolol lowered blood pressure and heart rate in both normotensive and hypertensive subjects, and reduced the frequency and duration of silent ischaemic episodes in hypertensive subjects. Plasma noradrenaline measured during silent ischaemia while the patients were resting was significantly higher than the control supine level without ischaemia. These findings suggest that noradrenergic hyperactivity may have a role in coronary vasoconstriction and that treatment which neutralizes sympathetic tone may be especially beneficial in treatment of silent ischaemia in hypertensive patients. PMID- 2563788 TI - Prevention of hepatitis B virus carrier state in infants according to maternal serum levels of HBV DNA. AB - 235 infants of HBeAg-carrier mothers in Hong Kong were assigned to four study groups. Groups I, II, and III received hepatitis-B (HB) vaccine at birth and at 1, 2, and 6 months. Group I also received seven monthly injections of HB immunoglobulin (HBIg), and group II received one HBIg injection at birth. Group III received vaccine only and group IV received placebo for both vaccine and HBIg. At the age of 3 years, all infants of the three treatment groups were significantly protected against the HB virus (HBV) carrier state compared with the placebo group (p less than 0.0001); the protective efficacy rates in groups I, II, and III were 87%, 80%, and 65%, respectively. At all times, group I was significantly better protected than group III. In groups III and IV, infants of mothers with serum HBV DNA levels of 5 pg/ml or above were at a significantly higher risk of acquiring the HBV carrier state than those whose mothers had HBV DNA levels below 5 pg/ml. This difference was not significant in groups given HBIg. Of the 183 infants who initially escaped HBV infection, 73 (40%) had transient and 8 (4%) chronic HBV infection between 6 and 36 months. Vaccinated infants who had actively formed anti-HBs remained well protected against the HBV carrier state. However, infants in groups I and II with no active anti-HBs response to vaccine became at risk for the HBV carrier state when the passively acquired anti-HBs antibodies had disappeared. HBIg should be included in HB vaccination schedules for all infants of HBeAg-positive mothers. PMID- 2563790 TI - Zidovudine in symptomless HIV infection. PMID- 2563789 TI - Diagnostic value of the apex beat. AB - 100 patients were examined without knowledge of other findings to assess the value of apex displacement as a sign of cardiomegaly; 25 had a radiographic cardiothoracic ratio greater than 50%. The apex was located in only half of the patients, palpability being influenced by frame size, percentage ideal body weight, and percentage body fat. By comparison with the cardiothoracic ratio, apex displacement beyond the midclavicular line as the diagnostic test for cardiomegaly had a specificity of 76%, a sensitivity of 59%, a positive predictive value of 59.4%, and a negative predictive value of 76.9%. Another clinical sign of cardiomegaly, apex more than 10 cm from the midsternal line, was more sensitive but even less specific. PMID- 2563791 TI - Anything to eat? PMID- 2563792 TI - An arranged marriage. PMID- 2563794 TI - Abdominojugular test. PMID- 2563793 TI - Bench surgery for renal tumours. PMID- 2563795 TI - Penicillin allergy in childhood. PMID- 2563796 TI - Late referral for biliary atresia--missed opportunities for effective surgery. AB - To assess whether clinicopathological features other than the age at operation influence prognosis after surgery for extrahepatic biliary atresia (EHBA) and to determine whether the age at referral has fallen since a previous survey, 50 consecutive cases with EHBA referred between February, 1985, and December, 1987, were reviewed. Liver or spleen size, liver function tests, or histological appearance of liver biopsy specimen before surgery were not predictive of outcome. The jaundice cleared up in 12 of 14 children operated on by age 8 weeks, but in only 13 of 36 operated on later. In 41 referral was delayed. All 25 children in whom surgery was successful are alive and well, while 13 of 25 with unsuccessful surgery have died, at a median age of 1 year. To improve the prognosis of infants with EHBA parents and health staff need a better awareness of the early clinical features of EHBA and of the necessity for prompt referral. Liver disease should be suspected in any infant jaundiced after 14 days of age. PMID- 2563797 TI - Individual and group hypnotherapy in treatment of refractory irritable bowel syndrome. AB - 33 patients with refractory irritable bowel syndrome were treated with four 40 minute sessions of hypnotherapy over 7 weeks. 20 improved, 11 of whom lost almost all their symptoms. Short-term improvement was maintained for 3 months without further formal treatment. Hypnotherapy in groups of up to 8 patients was as effective as individual therapy. PMID- 2563798 TI - Pharmacokinetics of recombinant human erythropoietin in patients on continuous ambulatory peritoneal dialysis. AB - To determine the optimum regimen for giving recombinant human erythropoietin (EPO) to patients on continuous ambulatory peritoneal dialysis (CAPD), the pharmacokinetics of single-dose EPO administered intravenously (120 U/kg), intraperitoneally (50,000 U), and subcutaneously (120 U/kg) was investigated. After intravenous administration serum EPO levels decayed exponentially from a peak of 3959 mU/ml, with a half-life of 8.2 h. 2.3% of the total intravenous dose was lost in the dialysate during the first 24 h. Peak serum EPO levels of 375 mU/ml at 12 h and 176 mU/ml at 18 h were attained following intraperitoneal and subcutaneous administration, respectively. The bioavailability of subcutaneous EPO (21.5%) was seven times greater than that of intraperitoneal EPO (2.9%). These results suggest that subcutaneous EPO represents the most satisfactory route of administration for CAPD patients. PMID- 2563799 TI - Indoor air pollution in developing countries and acute respiratory infection in children. PMID- 2563800 TI - Adverse effects of sunscreens in photosensitive patients. AB - Experimental and epidemiological evidence shows that the common photosensitive disorder of polymorphic light eruption is caused by the ultraviolet A component of sunlight. Sunscreens protect mainly against ultraviolet B; consequently they reduce sunburn and allow longer periods of exposure to the sun and to greater doses of ultraviolet A than would otherwise be possible. Patients with polymorphic light eruption who intend to obtain a tan by sunbathing should not, therefore, be treated with sunscreens which may worsen their rash, but should be advised to sunbathe without sunscreens for a shorter time. PMID- 2563801 TI - Bioavailability of aluminium. PMID- 2563802 TI - Abdominal incision. PMID- 2563803 TI - Alpha-linolenic acid. PMID- 2563804 TI - High rate of human papillomavirus type 16 infection in cytologically normal cervices. PMID- 2563805 TI - Adverse effects of lisinopril. PMID- 2563806 TI - Isotretinoin and eosinophilic pleural effusion. PMID- 2563807 TI - Hepatitis B and mental handicap. PMID- 2563808 TI - Lasegue's sign. PMID- 2563809 TI - Priapism in patient on tamoxifen. PMID- 2563810 TI - Maternal immune response and recurrent miscarriage. PMID- 2563811 TI - Retinal changes, gas bubbles, and diving. PMID- 2563813 TI - Hyponatraemia and haemolytic uraemic syndrome. PMID- 2563812 TI - Atlantoaxial instability, sport, and Down syndrome. PMID- 2563814 TI - Proguanil. PMID- 2563815 TI - Infectivity of unconventional viruses in dura mater. PMID- 2563816 TI - Cardiac arrhythmias and haemodialysis, bicarbonate or acetate? PMID- 2563817 TI - Chronic inflammatory arthropathy associated with HTLV-I. PMID- 2563818 TI - Seronegative neuroborreliosis. PMID- 2563819 TI - Audit and necropsy. PMID- 2563820 TI - Brother-to-sister transmission of measles after MMR immunisation. PMID- 2563821 TI - Medical research funding. PMID- 2563822 TI - Baldness and minoxidil advertising. PMID- 2563823 TI - Small round structured viruses and their spread. PMID- 2563824 TI - Xp21 DNA probe in diagnosis of muscular dystrophy and spinal muscular atrophy. PMID- 2563825 TI - First trimester transvaginal sonographic diagnosis of fetal anomalies. PMID- 2563826 TI - Transvaginal ultrasonography for diagnosis of placenta praevia. PMID- 2563827 TI - Vaginal endosonography in endometrial cancer. PMID- 2563829 TI - Hazard to bronchoscopists. PMID- 2563828 TI - Home intravenous diuretic therapy for patient with refractory heart failure. PMID- 2563830 TI - Health hazard from mercury soap. PMID- 2563831 TI - Immunological evaluation before liver transplantation in children. PMID- 2563832 TI - Fluoride and osteoblasts. PMID- 2563833 TI - Non-invasive measurement of cerebral blood flow during cardiopulmonary bypass. PMID- 2563834 TI - 4-Aminosalicylic acid enemas for ulcerative colitis. PMID- 2563835 TI - Congenital renal dysgenesis possibly due to captopril. PMID- 2563836 TI - Plasma-derived versus recombinant hepatitis B vaccines. PMID- 2563837 TI - Sitting down with the literature. PMID- 2563838 TI - Mianserin product licence. PMID- 2563839 TI - Echocardiography in diagnosis of aortic dissection. AB - In a multicentre study the accuracy of echocardiography was measured in 164 consecutive patients with suspected aortic dissection. The diagnosis was subsequently proven (82 patients) on the basis of transthoracic and transoesophageal echocardiography and additional diagnostic procedures, including computed tomography (CT), aortic angiography, and surgery and/or necropsy. The sensitivity and specificity of echocardiography were 99% and 98%, respectively, with positive and negative predictive values of 98% and 99%. For CT sensitivity was 83%, specificity 100%, and positive and negative predictive values 100% and 86%, respectively. For aortography sensitivity and specificity were 88% and 94%, and the positive and negative predictive values 96% and 84%, respectively. Echocardiography, including the transoesophageal route, can provide accurate diagnosis of aortic dissection within 15 min. Diagnostic difficulties occur only in a few type II dissections, when complementary diagnostic procedures, including CT or angiography, may be needed. To clarify branch involvement angiography is required. PMID- 2563841 TI - Abnormal jejunal potential difference in cystic fibrosis. AB - Transmucosal potential difference (PD) and intraluminal pressure were recorded from the same jejunal site in 15 healthy adult controls and 4 adults with cystic fibrosis. In the controls, runs of contractions were associated with wave-like changes in PD with, during phase III-like activity, a mean peak amplitude of -9.7 mV. Although there were no obvious differences in motor patterns, wave-like changes in PD were absent in patients with cystic fibrosis. Intraluminal boluses of 4 mg pilocarpine, or 0.1 mg prostaglandin E2, caused changes of -4.6 mV and 4.5 mV, respectively, in controls; these responses were not seen in patients with cystic fibrosis. There were no significant differences in basal PD and PD changes caused by altered concentrations of infused saline or glucose between patients and controls. PMID- 2563840 TI - Features of endothelial dysfunction in early diabetic nephropathy. AB - The release of tissue plasminogen activator (tPA) by vascular endothelial cells during exercise was studied in forty men with insulin-dependent diabetes. Three groups, matched for age and diabetes duration, were defined as: group I (n = 19), normal urinary albumin excretion (less than 30 mg/24 h); group II (n = 11), incipient diabetic nephropathy (30-300 mg albumin excreted per 24 h); and group III (n = 10), clinical diabetic nephropathy (more than 300 mg albumin excreted per 24 h). Nine non-diabetic men served as controls. The rise in tPA antigen with exercise was similar in the controls and group I but significantly smaller in groups II and III (p less than 0.01). The albumin transcapillary escape rate was significantly higher in groups II and III than in group I and normal controls (p less than 0.01). The basal plasma level of von Willebrand factor was higher in groups III (p less than 0.01) and II (difference not significant, p = 0.06) than in group I and normal controls. These findings suggest that insulin-dependent diabetic patients with only slightly raised urinary albumin excretion have general endothelial cell dysfunction or damage. It is not yet clear whether these changes are important in the pathogenesis of thrombosis and atherosclerosis in these patients. PMID- 2563842 TI - Distinction of Becker from limb-girdle muscular dystrophy by means of dystrophin cDNA probes. AB - The similarity in clinical features of X-linked Becker muscular dystrophy (BMD) and the autosomal recessive limb-girdle (LGD) type of adult muscular dystrophy makes differential diagnosis of the isolated male case difficult. DNA probes complementary (cDNA) to the Duchenne/Becker muscular dystrophy gene product, dystrophin, can detect molecular deletions in 60-70% of affected subjects. Thirty three patients with BMD or LGD (thirty isolated and three with an affected brother) were screened with a panel of cDNA probes for the whole dystrophin gene. Deletions were found in thirteen of eighteen (72%) patients with a diagnosis of BMD. Deletions were also found in four of the fifteen (27%) patients previously thought to have LGD, who were therefore reclassified as having BMD. All male patients with progressive muscular dystrophy of limb-girdle pattern should be routinely screened with these cDNA probes as a useful adjunct to their clinical diagnosis since the results have important implications for genetic counselling of affected families. PMID- 2563843 TI - Soluble interleukin-2 receptors in serum and bile of liver transplant recipients. AB - Soluble interleukin-2 receptors (IL2R) were measured as markers of lymphocyte activation in serum and bile of liver transplant recipients. Serum and biliary levels were significantly higher in patients with acute rejection than in those with other complications (serum p less than 0.0025, bile p less than 0.001) or stable grafts (both p less than 0.0001). Levels rose 24 h before rejection could be detected by conventional liver tests. Biliary levels were more specific and sensitive than serum levels for rejection. Local production of IL2R accounted for the high levels in bile; the bile to serum ratio of IL2R was greater than that of albumin in 16 of 18 patients with acute rejection. Serum levels were high early in the course of chronic rejection but fell as it progressed to end-stage. Measurement of soluble IL2R may have a role in the early diagnosis of acute rejection and in identifying patients with chronic rejection in whom further immunosuppression will provide no benefit. PMID- 2563844 TI - Secondary prevention of coronary disease with lipid-lowering drugs. PMID- 2563845 TI - Senile dementia of Alzheimer's type--normal ageing or disease? PMID- 2563846 TI - Brush cytology for colorectal cancer. PMID- 2563847 TI - Zinc fingers and vitamin D resistance. PMID- 2563848 TI - Population interventions reassessed. PMID- 2563849 TI - Health promotion and the compression of morbidity. PMID- 2563850 TI - Lyme disease: prevalence and clinical importance of Borrelia burgdorferi specific IgG in forestry workers. AB - 41 forestry workers, who had a high occupational risk of tick-bites, were screened for antibodies to Borrelia burgdorferi by ELISA and western blotting techniques, and questioned about possible symptoms of Lyme disease. Antibodies were detected in 10 of the 40 men who had been bitten by ticks. Definite symptoms of Lyme disease, in the form of erythema migrans, were reported by only 2 workers and none had a history of neurological illness. PMID- 2563851 TI - Ghee applications to the umbilical cord: a risk factor for neonatal tetanus. AB - A case-control study was done to elucidate risk factors for neonatal tetanus (NNT) in rural Pakistan; three controls were selected for each of the 69 cases. Factors commonly believed to be associated with NNT, such as the tool used to cut the umbilical cord and the method of preparing the tool, were not confirmed as risk factors. However, applying ghee to the wound stump was. New prevention opportunities deriving from the finding should be pursued aggressively while efforts to develop effective delivery systems to immunise mothers against tetanus continue, a task made difficult by sociocultural factors. PMID- 2563852 TI - Aluminium in infant formulas. PMID- 2563853 TI - Aluminium encephalopathy and Alzheimer's disease. PMID- 2563854 TI - Endogenous benzodiazepines in hepatic encephalopathy. PMID- 2563855 TI - Investigation of rectal bleeding. PMID- 2563856 TI - Listeriosis in Scotland, 1988. PMID- 2563857 TI - Life expectancy, Down syndrome, and dementia. PMID- 2563858 TI - Evolution of lactase persistence. PMID- 2563859 TI - Nasopharyngeal oxygen in children. PMID- 2563860 TI - Granulocyte-macrophage colony-stimulating factor for granulocyte defects of bone marrow transplant patients. PMID- 2563861 TI - Pain and cervical laser therapy. PMID- 2563862 TI - Family history of type 2 diabetes in schizophrenic patients. PMID- 2563863 TI - Incentive for measles, mumps, and rubella vaccination. PMID- 2563864 TI - Inherited deletion at Duchenne dystrophy locus in normal male. PMID- 2563865 TI - Excretion of platinum into breast milk. PMID- 2563866 TI - Young dyspeptic patients. PMID- 2563867 TI - Food irradiation. PMID- 2563868 TI - Israeli doctors and human rights. PMID- 2563869 TI - Reform of law on abortion. PMID- 2563870 TI - Morbidity of very-low-birthweight infants. PMID- 2563871 TI - Screening for inborn errors of purine synthesis. PMID- 2563872 TI - Listeria monocytogenes bacteraemia in AIDS. PMID- 2563873 TI - Specificities of HIV-1 antibodies in serum and in peripheral blood supernatants. PMID- 2563874 TI - Aerosolised pentamidine. PMID- 2563876 TI - SPECT and PET in epilepsy. PMID- 2563875 TI - Asthma epidemics and soybean in Cartagena (Spain) PMID- 2563877 TI - Gentamicin and antiseptic resistance in epidemic methicillin-resistant Staphylococcus aureus. PMID- 2563878 TI - Essential drugs in a European setting. PMID- 2563879 TI - Dietary guidelines for infants. PMID- 2563880 TI - Efficacy of salbutamol in chronic cough in childhood leukaemia. PMID- 2563881 TI - Stroke prevention and oestrogen replacement therapy. PMID- 2563882 TI - Transmission of hepatitis B from gynaecologist to patient. PMID- 2563883 TI - Toxicology of mussels contaminated by neuroexcitant domoic acid. PMID- 2563884 TI - Computed tomography in childhood acute lymphoblastic leukaemia. PMID- 2563885 TI - Surgical treatment of hypertrophic obstructive cardiomyopathy. PMID- 2563886 TI - Clonidine for short stature. PMID- 2563887 TI - Microbiology at the crossroads. PMID- 2563888 TI - Abdominal incision. PMID- 2563889 TI - Circulating anti-elastase in systemic lupus erythematosus. PMID- 2563890 TI - Adverse reactions to carbamazepine managed by desensitisation. PMID- 2563891 TI - Uninsured pharmacists and illegible prescriptions. PMID- 2563892 TI - HIV infection and drug misusers. PMID- 2563893 TI - Effects of ingestion of glucose on GH and TSH secretion: evidence for stimulation of somatostatin release from the hypothalamus by acute hyperglycemia in normal man and its impairment in acromegalic patients. AB - Ingestion of glucose is known to induce suppression of GH secretion in normal subjects and this phenomenon is often absent in acromegalic patients. To clarify the mechanism of GH suppression in acute hyperglycemia in normal subjects and disturbed GH response in acromegalic patients, the effects of acute hyperglycemia on plasma GH and TSH levels were examined in normal subjects and acromegalic patients. Plasma GH levels were significantly lowered 45-60 min after ingestion of 75 g glucose and elevated at 210 and 240 min in nine normal subjects. Plasma TSH levels were also significantly lowered between 45 and 120 min after ingestion; levels then gradually rose. Subcutaneous administration of 50 micrograms SMS 201-995, a long acting somatostatin analog, lowered plasma TSH levels in both normal subjects and acromegalic patients, and there was no significant difference in the degree of decrease in plasma TSH levels between the normal subjects and patients. These results, taken together with several reports that somatostatin suppresses TSH secretion as well as GH secretion, suggest that acute hyperglycemia stimulates somatostatin release from the hypothalamus, thus causing inhibition of GH and TSH secretion. However, in ten acromegalic patients, only two showed suppression of plasma GH levels to below 50% of basal level and the degree of suppression of TSH secretion was significantly less than in normal subjects in the glucose tolerance test. It is, therefore, suggested that somatostatin release in response to acute hyperglycemia is impaired in most acromegalic patients and that this abnormality may be one of causes for the absence of the normal GH response to acute hyperglycemia in this disorder. PMID- 2563894 TI - Drugs for psychiatric disorders. PMID- 2563895 TI - Oral hypnotic drugs. PMID- 2563896 TI - Comparison of invasive and conservative strategies after treatment with intravenous tissue plasminogen activator in acute myocardial infarction. Results of the thrombolysis in myocardial infarction (TIMI) phase II trial. AB - We treated 3262 patients with intravenous recombinant tissue plasminogen activator (rt-PA) within four hours of the onset of chest pain thought to be caused by myocardial infarction. Of these patients, 1636 were then randomly assigned to treatment according to an invasive strategy consisting of coronary arteriography 18 to 48 hours after the administration of rt-PA, followed by prophylactic percutaneous transluminal coronary angioplasty (PTCA) if arteriography demonstrated suitable anatomy; 1626 patients were randomly assigned to treatment according to a conservative strategy, as part of which arteriography and PTCA were to be performed only in patients with spontaneous or exercise induced ischemia. In the group assigned to the invasive strategy, PTCA was attempted in 928 of the 1636 patients (56.7 percent); the procedure was anatomically successful in 93.3 percent. In the group assigned to the conservative strategy, 216 patients (13.3 percent) underwent clinically indicated PTCA within 14 days of the onset of symptoms. Reinfarction or death within 42 days, the primary end point, occurred in 10.9 percent of the group assigned to the invasive strategy and in 9.7 percent of those assigned to the conservative strategy (P not significant). There were no significant differences between the two groups in the ejection fraction at rest or during exercise, either at hospital discharge or six weeks after randomization. Eleven of 582 patients (1.9 percent) who received 150 mg of rt-PA and 15 of 2952 patients (0.5 percent) who received 100 mg of rt-PA had intracranial hemorrhage. A subgroup of 1390 patients who were eligible for short-term intravenous beta-blockade were randomly assigned to receive 15 mg of intravenous metoprolol immediately, followed by oral metoprolol, or oral metoprolol begun on day 6. The ejection fraction and the incidence of death in the two groups were similar during the hospital period. Total mortality within the first 6 days and at 42 days was also similar. However, in the group that received intravenous metoprolol, 16 patients had nonfatal reinfarctions and 107 patients had recurrent ischemic episodes by six days after entry into the study, as compared with 31 and 147 patients, respectively, among those randomly assigned to deferred oral beta-blockade (P = 0.02 and P = 0.005, respectively); the latter comparison was considered statistically significant according to the study criteria.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2563897 TI - Viral proliferating cell nuclear antigen. PMID- 2563898 TI - Characterization of the yeast HSP60 gene coding for a mitochondrial assembly factor. AB - The hsp60 protein isolated from the protozoan Tetrahymena thermophila is induced in response to heat stress and is a member of an immunologically conserved family represented in Escherichia coli and in mitochondria of plants and animals. We report here the cloning and characterization of a nuclear gene, HSP60, which codes for the hsp60 homologue from the yeast Saccharomyces cerevisiae. Nucleotide sequence analysis revealed that yeast hsp60 is related to the groEL protein of E. coli and the RUBISCO-binding protein (RBP) of chloroplasts. HSP60 was found to be the genetic locus of the conditional-lethal mutation described by Cheng et al., which at non-permissive temperature is defective in the assembly of several different multisubunit complexes in mitochondria. These data are consistent with the hypothesis that the groEL-related proteins serve an evolutionarily conserved function as accessory factors facilitating the folding and/or association of individual subunits of multimeric protein complexes. PMID- 2563899 TI - Guanylate cyclases send out new signals. PMID- 2563900 TI - A membrane form of guanylate cyclase is an atrial natriuretic peptide receptor. AB - Atrial natriuretic peptide (ANP) is a polypeptide hormone whose effects include the induction of diuresis, natriuresis and vasorelaxation. One of the earliest events following binding of ANP to receptors on target cells is an increase in cyclic GMP concentration, indicating that this nucleotide might act as a mediator of the physiological effects of the hormone. Guanylate cyclase exists in at least two different molecular forms: a soluble haem-containing enzyme consisting of two subunits and a non-haem-containing transmembrane protein having a single subunit. It is the membrane form of guanylate cyclase that is activated following binding of ANP to target cells. We report here the isolation, sequence and expression of a complementary DNA clone encoding the membrane form of guanylate cyclase from rat brain. Transfection of this cDNA into cultured mammalian cells results in expression of guanylate cyclase activity and ANP-binding activity. The ANP receptor/guanylate cyclase represents a new class of mammalian cell-surface receptors which contain an extracellular ligand-binding domain and an intracellular guanylate cyclase catalytic domain. PMID- 2563901 TI - Production of chimaeric mice containing embryonic stem (ES) cells carrying a homoeobox Hox 1.1 allele mutated by homologous recombination. AB - Several mouse gene families related to Drosophila developmental control genes and containing a homoeobox, a paired box or a finger domain, have been cloned and structurally analysed. On the basis of structural similarities to the Drosophila genes and of their spatially and temporally restricted expression patterns during mouse embryogenesis, it has been proposed that these mammalian genes also are involved in the control of development. To elucidate the function of homoeobox genes by genetic means, mouse mutants must be generated. We have developed a technique for mutagenesis in vivo and have used it to mutate the homoeobox Hox 1.1 gene. In vivo mutagenesis was achieved through homologous recombination between an endogenous Hox 1.1 allele and a microinjected mutated gene in pluripotent embryonic stem (ES) cells. Mutant cells were identified by means of the polymerase chain reaction (PCR) and mutant clones were used to generate chimaeric mice. Because the homologous recombination event is formally a gene conversion event and no selection is required to screen for cells carrying the mutated allele, in vivo mutagenesis allows specific alterations in the target sequence to be made without the introduction of any other sequences. PMID- 2563902 TI - Production of a mutation in mouse En-2 gene by homologous recombination in embryonic stem cells. AB - A full understanding of the function of genes that control developmental events can be obtained only by a combination of molecular and mutational analysis. One putative developmental gene is the mouse engrailed-like gene En-2, which was isolated by virtue of its extensive homology to Drosophila engrailed, which contributes to the control of segmentation in the developing insect. Our hybridization analysis in situ has revealed that expression of En-2 is restricted to a specific domain of the developing central nervous system from 8 days of development on, indicating a role for the gene in establishing spatial domains in the brain. Unfortunately no En-2 mutations are available to elucidate further its function in development. To this end, we report here the isolation of three pluripotent embryonic stem cell lines in which one copy of the homoeobox containing gene, En-2, has been altered by homologous recombination. PMID- 2563903 TI - Intraglomerular deposition of coagulation-fibrinolysis factors and a platelet membrane antigen in various glomerular diseases. AB - The intraglomerular location of coagulation-fibrinolysis factors (CFF) and a platelet membrane antigen (glycoprotein IIb-IIIa; GPIIb-IIIa) was determined in 101 patients with various glomerular diseases. Renal biopsy specimens were examined by immunofluorescence microscopy, using antisera against fibrinogen/fibrin reactive antigen (FRA), cross-linked fibrin degradation products (XL-FDP), fibronectin (FN), factor XIII-subunit a (F-XIIIa), plasminogen (Plg), alpha 2-plasmin inhibitor (alpha 2-PI) and GPIIb-IIIa. Intraglomerular deposits of the CFF were found at high rates in patients with IgA glomerulonephritis (GN), membranous nephropathy (MN) and lupus GN. The coexistence of deposits of these factors was ascertained by the double-staining method. The deposition rates of XL-FDP and GPIIb-IIIa were very low in patients with minimal-change nephrotic syndrome and focal glomerulosclerosis. Some cases of diabetic glomerulosclerosis (DGS) showed CFF deposition. FRA deposits associated with F-XIIIa and FN may indicate the presence of the cross-linked fibrin. Furthermore, the presence of Plg deposits together with alpha 2-PI and XL FDP suggests the deposition of fibrin followed by fibrinolysis, but not of fibrinogen, and the coexistence of GPIIb-IIIa suggests the involvement of platelets in the reactions. These studies provide evidence that stabilized fibrin deposition with subsequent fibrinolysis and platelet activation take place in glomeruli in a fairly large proportion of patients with IgA GN, MN and lupus GN and in some cases of DGS. PMID- 2563904 TI - Long-term potentiation in frontal cortex: role of NMDA-modulated polysynaptic excitatory pathways. AB - The present study examined the role of N-methyl-D-aspartic acid (NMDA) receptors in synaptic plasticity in regular-spiking cells of rat frontal cortex. Intracortical stimulation, at levels subthreshold for elicitation of action potentials, evoked a late excitatory postsynaptic potential (EPSP) in layer II III neurons that was sensitive to the selective NMDA antagonist D-2-amino-5 phosphonovaleric acid (APV). This late EPSP showed marked short-term frequency dependent depression, suggesting that it is polysynaptic in origin. Polysynaptic late EPSPs were selectively enhanced following high-frequency stimulation. This sustained increase in synaptic efficacy, or long-term potentiation, was expressed in regular spiking cells and appeared to result from activation of NMDA receptors on excitatory interneurons. These data demonstrate the existence of an NMDA modulated polysynaptic circuit in the neocortex which displays several types of use-dependent plasticity. PMID- 2563905 TI - Ontogeny of glutamate accumulating activity in rat brain synaptic vesicles. AB - The relationship between the ontogeny of the vesicular glutamate uptake system and synaptogenesis in rats was investigated. For this purpose we have developed a simplified procedure for the preparation of crude synaptic vesicles which are sufficiently pure to demonstrate a highly ATP-dependent glutamate uptake. ATP dependent glutamate uptake into synaptic vesicles was found to increase dramatically starting on postnatal day 10 and reaching a maximum on day 30 (76 +/ 40 and 657 +/- 40 pmol/mg protein/10 min, respectively), correlating well with the active period of synaptogenesis. Stimulation of uptake by chloride also developed in parallel with the vesicular glutamate uptake. In contrast, combined non-ATP-dependent uptake and non-specific binding remained constant (21 +/- 6 pmol/mg protein/10 min). This development of vesicular glutamate uptake during the period of synaptogenesis supports the notion that synaptic vesicles play an important role in glutamate synaptic transmission. PMID- 2563906 TI - Synthesis of glutamate and glutamine in dibutyryl cyclic AMP-treated astrocytes. AB - The relative contributions of radioactively labeled fatty acids and glucose to synthesis of glutamate and glutamine were compared in native and dibutyryl cyclic AMP (diBcAMP)-treated primary rat astrocytes. The intracellular specific activities of glutamate and glutamine were 10-fold greater than the specific activities of aspartate or alanine. Butyrate, octanoate and palmitate were equally as effective as precursors for glutamate and glutamine while glucose was 50% as effective as the fatty acids. The specific activity of glutamate and glutamine were identical in the absence of diBcAMP. In diBcAMP treated cells the specific activity of glutamine was greater than that of glutamate when octanoate and palmitate were the labeled precursors. This suggests that cultured astrocytes preferentially utilize free fatty acids for glutamate/glutamine synthesis and that diBcAMP-treated astrocytes contain more than one glutamate compartment. PMID- 2563907 TI - Ontogeny of tyrosine hydroxylase-positive but dopamine beta-hydroxylase-negative neuron-like cells in the pineal gland of golden hamsters. AB - The ontogeny of tyrosine hydroxylase (TH)-positive but dopamine beta-hydroxylase (DBH)-negative neuron-like cells in the pineal gland of golden hamsters was investigated by double immunostaining. These cells first appeared on the 6th postnatal day and thereafter increased in number. On the other hand, TH- and DBH positive nerve fibers, probably originating from the superior cervical ganglion, were already present in the pineal gland at birth. PMID- 2563908 TI - Tyrosine hydroxylase-like immunoreactive neurons in the striatum of the rat. AB - Striatal neurons exhibiting tyrosine hydroxylase-like immunoreactivity (TH-LI) were found in the adult rat. Most of these neurons had a cell body with a 10-20 microns diameter and several spiny dendrites. The number of striatal neurons with TH-LI was increased on the side ipsilateral to an electrothermic or 6 hydroxydopamine-induced lesion which had been placed in the regions around the substantia nigra. PMID- 2563909 TI - Multiple voltage-sensitive K+ channels regulate dendritic excitability in cerebellar Purkinje neurons. AB - Ionic conductances present in the dendritic region of the cerebellar Purkinje neuron were studied using the single-channel and whole-cell recording methods. Several types of voltage-sensitive K+ channels including a Ca2+ activated K+ channel were found to be a prominent components of the dendritic membrane. All patches studied contained K+ channel types and most patches contained more than one K+ channel type. In cell attached recordings, K+ channel activity was associated with the late phase of spontaneous action potentials suggesting a functional relationship. These data demonstrate that voltage-sensitive ion channels contribute to dendritic excitability and suggest that the transduction and integration of synaptic signals may involve both active and passive ionic conductances. PMID- 2563910 TI - [Rapid care after myocardial infarction crucial for the prognosis]. AB - Treatment in cases of acute myocardial ischaemia is currently undergoing dynamic changes. Several studies, involving thousands of patients with acute cardiac infarction, have shown beneficial effects of beta blockers, thrombolysis, and treatment with platelet aggregation inhibitors. Current research is focused on the definition of subgroups among those with acute myocardial ischaemia who are best served by a particular treatment or combination of treatments. PMID- 2563911 TI - [Role of the serum level of mitochondrial aspartate aminotransferase as marker of alcoholic intoxication in cirrhotic patients]. AB - The purpose of this study was to assess the diagnostic value of serum mitochondrial aspartate aminotransferase activity (mAST) and of the mitochondrial aspartate aminotransferase/total aspartate aminotransferase ratio (mAST/t AST) as markers of chronic alcoholism in cirrhotic patients. Sixty-three hospitalized cirrhotic patients (35 drinkers, 28 abstainers) were investigated. Ninety-six per cent of abstainers had normal values of mAST activity, while 89 per cent of drinkers had high levels of mAST activity. Cirrhotic patients were better divided into drinkers and abstainers by mAST activity (92 per cent) than by mean globular corpuscular volume (MCV) (81 per cent, NS) or by gamma-glutamyltransferase activity (GGT) (75 per cent, P less than 0.01). When the hospital costs of these markers were taken into account, MCV had a better "quality/price" ratio (Q/P) defined as diagnostic value/mean hospital cost (Q/P = 2.5) than MCV plus GGT (diagnostic value 61 per cent, Q/P = 1.2). The measurement of mAST activity in patients with high MCV value or with discrepancy between MCV and GGT values increased the diagnostic value of the other laboratory measurement to 89 per cent, but at a higher cost (Q/P = 0.8). Mitochondrial AST activity is a sensitive and specific marker of chronic alcoholism in cirrhotic patients. However, owing to its high cost, it should be proposed as a second marker after MCV and GGT. PMID- 2563912 TI - Facilitation as a tool to study the entry of calcium and the mechanism of neurotransmitter release. AB - We have shown the usefulness of using facilitation as an indirect tool for measuring release-related processes that cannot be measured directly. Most of the findings obtained by measuring facilitation have been verified (by various groups) by direct measurements in systems where such measurements could be carried out. This provides reassurance that the methodology is sound. Using facilitation one can gain insight into numerous processes that together govern the dependence of release on the intracellular calcium concentration C. The physiological conclusions have been listed in the text. We reiterate some of these conclusions here, in order to emphasize certain additional matters. For 20 years it has been known that release is a saturating cooperative function of the extracellular Ca2+ concentration Ce (Dodge and Rahamimoff, 1967). Yet several questions remained open, such as whether this behavior in fact reflected the dependence of the release on the intracellular Ca2+ concentration C, of entry on Ce, or of a combination of these individual possibilities. We have found, using short-term facilitation as the investigatory tool, that release is a saturating cooperative function of C. It is obvious that saturation will eventually take place as C increases. What is important to emphasize is that the saturation occurs at physiological values of C. Such values typically correspond to the amount of Ca2+ that enters following only one or very few pulses. Various aspects of facilitation F can be used to characterize the processes responsible for the removal of the calcium that enters the nerve terminal. Here we emphasized the role of the duration of F, but information can also be obtained by examining other aspects of facilitation (Parnas and Segel, 1980; H. Parnas et al., 1982; I. Parnas et al., 1982a). The principal conclusion is that removal shows saturation kinetics.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563913 TI - Neurochemical transmission and the sodium-pump. PMID- 2563914 TI - The limits of medicine: women's perception of medical technology. AB - This paper develops an analysis of women's perceptions of medical technology and the elements which shape them, and then draws out the implications for medicine and the and the medicalization thesis. In the first part of the paper we outline the macro-theoretical debates about medicalization and the role of medical technology in this process, and the consequences for those who use health care. The implications for women are given particular attention as they have a higher level of contact with health care than men. We then evaluate the arguments of these macro-theorists against evidence from two ethnographic studies, concerning women patients' and their doctors' attitudes to the use of minor tranquillizers and women's evaluations of medicine and medical practice. This provides a basis for questioning some of the assumptions of the macro-theorists regarding the social relations of medical technology and the medicalization thesis. PMID- 2563915 TI - Hypertension, alcohol and cations in urban black and coloured South Africans. AB - Epidemiological evidence shows a positive relationship between alcohol consumption and blood pressure. High alcohol intake and hypertension are common in urban South African men. Relationships between mean arterial pressure (MAP), serum gamma-glutamyltransferase (GGT) as an index of alcohol intake, age, mass and levels of cations in the serum and erythrocytes were investigated in normotensive and hypertensive black and coloured men. Serum levels of potassium, magnesium and calcium and the red blood cell magnesium level were found to be significantly decreased in the black hypertensives. Serum GGT was equally raised in normotensive and hypertensive blacks and was positively correlated with systolic blood pressure in the hypertensive subjects. The coloured hypertensives were heavier, older and had significantly higher serum GGT levels than their normotensive counterparts. Serum GGT was positively correlated with MAP in the coloured subjects. No consistent relationships were found between GGT and blood cations. These data suggest that in younger black subjects alcohol is associated with systolic blood pressure only once hypertension has developed. Other factors, such as cations, may be more important than alcohol in the pathogenesis of hypertension in this group. Alcohol consumption is an important risk factor in coloured hypertensives. PMID- 2563917 TI - Biochemical and cellular changes in bronchoalveolar lavaged samples from rats after inhalation of mosquito-coil smoke. AB - A group of 30 female albino rats were exposed to mosquito-coil smoke, 8 hours a day, 6 days per week, for 6 months. Another group which was exposed to air served as control. At the end of the experiment, the enzyme activities, total protein and lecithin contents as well as cellular responses in the lung lavage between the control and smoke-exposed rats were compared. Morphological observations using scanning and transmission electron microscopy demonstrated that the alveolar macrophages of smoke-exposed rats lost their typical ruffled membranes. They possessed small cytoplasmic processes on their smooth cell surfaces, small particles in phagolysosomes and mitochondria with a very electron-dense matrix. The levels of total protein and lecithin and the activities of lactate dehydrogenase, acid phosphatase and beta-glucuronidase in the lung-lavage fluid of smoke-exposed rats were significantly (P less than 0.05) higher than those of the controls. Increases (P less than 0.05) of serum enzymes, including lactate dehydrogenase, aspartate aminotransferase, isocitrate dehydrogenase and aldolase, indicated damage of liver tissues, but the levels of serum urea and urea nitrogen remained at the control levels implying normal functions of the kidneys of the mosquito-coil smoke-exposed rats. The level of serum tri-iodothyronine also increased significantly (P less than 0.05), but thyroxine remained at the control level. PMID- 2563916 TI - Differential sparing of somatostatin-neuropeptide Y and cholinergic neurons following striatal excitotoxin lesions. AB - We previously found that quinolinic acid striatal excitotoxin lesions result in a relative sparing of somatostatin and neuropeptide Y neurons. In the present study we examined dose-response effects of excitotoxins acting at the three subtypes of glutamate receptors: N-methyl-D-aspartate (AA1), quisqualate (AA2), and kainic acid (AA3). Concentrations of both somatostatin-like immunoreactivity (SLI) and neuropeptide a Y-like immunoreactivity (NPYLI) were compared with those of substance P-like immunoreactivity (SPLI) and GABA. Kainic acid (AA3), quisqualic acid (AA2), and AMPA (AA2) resulted in dose-dependent reductions in all four neurochemical markers examined, while N-methyl-D,L-aspartate (AA1) and quinolinic acid (AA1) resulted in relative sparing of SLI and NPYLI. At doses of each excitotoxin which resulted in comparable 50% reductions in both GABA and SPLI only N-methyl-D,L-aspartate and quinolinic acid had no significant effect on concentrations of SLI and NPYLI. The relative sparing of somatostatin neuropeptide Y neurons was confirmed histologically by using histochemical staining for NADPH-diaphorase neurons combined with either Nissl stains, or immunohistochemical staining for enkephalin. Lesions with N-methyl-D-aspartate agonists resulted in preferential sparing of NADPH-diaphorase neurons while these neurons were more vulnerable than other neurons to kainic acid or AMPA. Choline acetyltransferase neurons were relatively spared, as compared with other neurons, by agents acting at all three glutamate receptor subtypes. N-methyl-D,L-aspartate lesions were blocked with MK-801, while there was no effect on quisqualic acid or kainic acid lesions. The relative sparing of somatostatin-neuropeptide Y neurons following striatal excitotoxin lesions with N-methyl-D-aspartate (AA1) agonists probably reflects a paucity of AA1 receptors on these neurons. Since these neurons are also spared in Huntington's disease, excitotoxins acting at the N methyl-D-aspartate (AA1) site provide an improved neurochemical model of this illness. PMID- 2563918 TI - Role of neurotransmitters in fenitrothion-induced aggressive behaviour in normal and lesioned rats. AB - Normal and brain-lesioned (amygdala, septal and nigral regions) rats exposed to fenitrothion (10 mg/kg) for 15 subsequent days showed elevated foot-shock fighting behaviour (septal and nigral lesioned rats). Treated animals failed to perform the rotorod test and showed decreased locomotor activity followed by convulsions. The brain-lesioned rats treated with 10 mg/kg fenitrothion showed a marked decrease in the level of norepinephrine (NE) and serotonin (5-HT). PMID- 2563919 TI - Platelet levels of glutamate and aspartate in normal subjects. PMID- 2563920 TI - Transmembrane signalling via the T cell antigen receptor heterodimer and the CD2 antigen. A synergistic pathway for activation of T cells. AB - T cell surface proteins involved in transmembrane signalling resulting in the activation of T cells were investigated utilizing as probes monoclonal antibodies directed at T cell surface antigens. Here we report that mAbs that react with a framework determinant of alpha/beta heterodimer of T cell receptor for antigen, anti-TCR-1, and those with the SRBC-binding epitope of the CD2 antigen, OKT11, are synergistic in promoting T cell proliferation. The proliferative response is dependent upon crosslinking of anti-TCR-1 and OKT11, and is associated with a significant increase in the concentration of intracellular free calcium in T cells. Moreover, EGTA and a direct (staurosporine) or a competitive (1-[5 isoquinolinylsulfonyl]-2-methyl piperazine) inhibitor of protein kinase C prevents T cell proliferation accomplished with crosslinked anti-TCR-1 and OKT11. Our findings, in addition to demonstrating the synergism between the signals initiated via the T cell receptor for antigen and the CD2 antigen, suggest a role for calcium and protein kinase C in the transduction of signals generated with crosslinked anti-TCR-1 and OKT11. PMID- 2563921 TI - [Cryopreservation of hemopoietic stem cells in autologous bone marrow transplantation]. AB - Autologous bone-marrow transplantation (AKMT) is employed to shorten the period of aplasia following aggressive radiotherapy and chemotherapy. Deep freezing of haemopoietic stem cells (HS) must be undertaken in this connection. In order to achieve the greatest survival of HS, the following requirements must be fulfilled: 1) The use of acid citrate dextrose as an anticoagulation agent is recommended. 2) The best results are obtained if a concentrate of mononuclear cells is deep frozen. 3) A cryoprotective agent must be employed in order to avoid injurious osmotic gradients through the cell membrane. Ten percent di methyl sulphoxide has been most tested and is most suitable. Cell concentrations before freezing must be 1-5 x 10(7) nuclear cells (NBC) ml. 4) Cells must be frozen in a homogenous 3-4 mm thick layer in special bags made of temperature resistant material. 5) Controlled freezing at a rate of between 1 and 2 degrees C/minute is employed. The heat yield at the eutechnical point must be eliminated. 6) Storage must be undertaken at temperatures lower than -140 degrees C. Bone marrow should not be employed after storage for three years. 7) Agar culture is employed as a screening test to demonstrate if an adequate number of cells are available. 8) The minimum dosage at AKMT is assessed as 1-7 x 10(7) NBC/kg. 9) The threshold dosage at AKMT is assessed to be 5-10 x 10(7) NBC/kg. 10) Thawing must be done rapidly. 11) The infusion should be concluded at the latest 15 minutes after thawing.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563922 TI - [Testicular retention]. PMID- 2563923 TI - Recent studies on the effects of beta blockers on blood lipid levels. PMID- 2563924 TI - The Nifedipine-Total Ischemia Awareness Program: a national survey of painful and painless myocardial ischemia including results of antiischemic therapy. AB - The Nifedipine-Total Ischemia Awareness Program was designed to evaluate the prevalence, prognostic implications and effect of therapy on painful and painless myocardial ischemic episodes in a nationwide study of patients with angina pectoris. Three hundred forty-eight patients with at least 2 anginal attacks/week while taking antianginal medications were enrolled at 53 participating centers between September 1, 1986 and March 31, 1988; 312 of the 348 patients formed the study group, while 36 patients formed the control group. At least 1 episode of ST segment depression during two 48-hour periods of Holter monitoring was present in 136 of the 312 patients in the study group. In these 136 patients, there was a total of 372 episodes of ST-segment depression, of which only 69 (18%) were painful; 85% of the 136 patients had either painless episodes only or both painless and painful episodes. Despite apparently adequate antianginal therapy, 48 patients had greater than or equal to 3 episodes of ST-segment depression/48 hours of ambulatory electrocardiographic monitoring, and 38 patients greater than 60 minutes of ST-segment depression. After nifedipine was administered, there was a 23% reduction in the mean number of episodes of ST-segment depression (2.7 +/- 0.3 to 2.1 +/- 0.2, p less than 0.01). The most pronounced effects were found in the 48 patients with greater than or equal to 3 episodes of ST-segment depression and the 38 patients with greater than or equal to 60 minutes of total ischemic time.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563925 TI - Hypnotherapy as an adjunct to narcotic analgesia for the treatment of pain for burn debridement. AB - This paper presents a hypnotherapeutic intervention for controlling pain in severely burned patients while they go through dressing changes and wound debridement. The technique is based on Barber's (1977) Rapid Induction Analgesia (RIA) and involves hypnotizing patients in their rooms and having their nurses provide posthypnotic cues for analgesia during wound cleaning. Five subjects who underwent hypnotherapy showed reductions on their pain rating scores (Visual Analogue Scale) relative to their own baselines and to the pain curves of a historical control group (N = 8) matched for initial pain rating scores. Although the lack of randomized assignment to experimental and control groups limited the validity of the results, the findings provide encouraging preliminary evidence that RIA offers an efficient and effective method for controlling severe pain from burns. PMID- 2563926 TI - H2 antagonists in the treatment of reflux esophagitis: a critical analysis. AB - To ascertain through clinical and endoscopic assessment, the clinical benefits of using H2 antagonists to treat patients with reflux esophagitis, we researched the English language literature using Index Medicus (1978-87) and bibliographic resources of textbooks. We selected 10 of 20 articles originally reviewed, which specifically addressed the stated purpose. Forty-three percent to 63% of patients treated with H2 antagonists improved symptomatically, compared with 9% to 35% of patients receiving placebo. Sixty percent to 88% of patients receiving H2 antagonists improved endoscopically, compared with 26% to 28% of patients receiving placebo. H2 antagonists are an effective treatment for reflux esophagitis. They cause a symptomatic and endoscopic improvement, mainly in moderate-to-severe erosive esophagitis. PMID- 2563927 TI - Effect of epoxyeicosatrienoic acids on growth hormone release from somatotrophs. AB - Growth hormone secretion was stimulated in vitro by products of arachidonic acid epoxygenase, the epoxyeicosatrienoic acids. 5,6-Epoxyeicosatrienoic and 14,15 epoxyeicosatrienoic acid stimulated growth hormone release from an enriched population of somatotrophs (approximately 85%) by twofold. Inhibition of arachidonic acid metabolism by indomethacin did not affect growth hormone releasing hormone stimulation of growth hormone release. In contrast, pretreatment of somatotrophs with an 11,12-isonitrile analogue of arachidonic acid that inhibits arachidonic acid epoxygenase, resulted in a 20-25% inhibition of growth hormone-releasing hormone-stimulated growth hormone release. 14,15 Epoxyeicosatrienoic acid stimulated a concentration-dependent increase (twofold) in the cytoplasmic concentration of adenosine 3',5'-cyclic monophosphate (cAMP) in the somatotrophs. 14,15-Epoxyeicosatrienoic acid also rapidly increased the intracellular free calcium concentration in somatotrophs from resting levels (approximately 80 nM) to greater than 250 nM. Growth hormone-releasing hormone increased the free intracellular calcium to 160-180 nM. Preincubation of somatotrophs with somatostatin inhibited growth hormone-releasing hormone stimulated growth hormone secretion, cAMP accumulation, and 14,15 epoxyeicosatrienoic acid stimulated cAMP accumulation. These data are suggestive that the epoxyeicosatrienoic acids may have a role in the secretion of growth hormone. PMID- 2563928 TI - Linkage between somatostatin and acid secretion: evidence from use of pertussis toxin. AB - Pertussis toxin was used to examine the functional linkage between somatostatin and acid secretion and the mode of action of somatostatin at the cellular level in the isolated luminally perfused mouse stomach. Pretreatment of the stomach with pertussis toxin (125-1,250 ng/ml) for 60 min 1) caused a significant twofold increase in histamine-stimulated acid secretion (from 42 +/- 7 to 82 +/- 12 nmol/min; P less than 0.01) but not pentagastrin-stimulated secretion and 2) blocked the inhibitory effect of somatostatin on basal and histamine-stimulated acid secretion but not on pentagastrin-stimulated acid secretion. The ability of pertussis toxin to reverse selectively the inhibitory effect of somatostatin on histamine-stimulated acid secretion is consistent with the ability of pertussis toxin to inactivate a guanine nucleotide binding protein, which couples somatostatin receptors to inhibition of adenylate cyclase; histamine, but not gastrin, stimulates acid secretion via activation of adenylate cyclase. Secretagogue-stimulated acid secretion was accompanied by a parallel increase in somatostatin secretion that is largely determined by luminal acidity. The augmentation of histamine-stimulated acid secretion after treatment with pertussis toxin implied that the concomitant increase in somatostatin secretion is coupled to acid secretion and acts to attenuate it. The results confirm the role of gastric somatostatin as a paracrine regulator of acid secretion. PMID- 2563929 TI - The Shanghai 800: prevalence of tardive dyskinesia in a Chinese psychiatric hospital. AB - The authors found that 73 (8.4%) of 866 patients with chronic schizophrenia in the Shanghai Psychiatric Hospital who had been treated with neuroleptics had tardive dyskinesia. This low prevalence rate is possibly ascribable to the use of relatively low doses of neuroleptics. PMID- 2563930 TI - Underrecognized and underresearched side effects of neuroleptics. PMID- 2563931 TI - [Comments on the paper by G. Steiner et al. Effects and side effects of somatostatin]. PMID- 2563932 TI - Neuropsychological side effects of beta-blockers. AB - Fifty-five studies of cognitive side effects of beta-blockers were reviewed. Many of the studies were limited by small sample size, use of drugs out of the range of normal administration, failure to control for known confounders for neuropsychological evaluation, or lack of a crossover design. As a result, one needs caution in drawing conclusions on this topic. Nevertheless, given the widespread use of beta-blockers and the frequent allegations about their relative side effect profiles, it is appropriate to summarize the results of these disparate studies. Across all beta-blockers and all cognitive domains, the drugs led to improved functioning in 16% of observations and worsened functioning in 17%, with no significant effect in the rest. Memory, learning, and abstraction have been studied less frequently. The perceptual motor cognitive domain was the most widely studied and was frequently affected by these drugs. There was no consistent evidence of a trend for lipophilic drugs to impair this domain more than lipophobic drugs. There is some evidence that these drugs have a positive effect on complex task performance. The drugs also seem to increase sedation; however, in these studies there was no evidence for a differential effect across lipophilic vs lipophobic drugs. PMID- 2563933 TI - Regional cerebral blood flow in depression and mania. AB - Forty-three depressed inpatients, referred for electroconvulsive therapy, and 30 manic patients were examined with clinical ratings and regional cerebral blood flow (rCBF) determinations. The depressed patients were mainly medication free, while most of the manic patients were medicated. Both patient groups showed a normal cerebral blood flow level and regional distribution compared with age- and sex-matched normal controls. In the depressed group and especially in the unipolar subgroup, a significant positive relationship was found between the mean rCBF and symptoms of depression and cognitive dysfunction. Eighteen of the depressed and 18 of the manic patients were reexamined in a euthymic state following treatment and recovery. Only minor and statistically nonsignificant flow changes were found in connection with the clinical improvement. In the manic patients, a significant negative relationship was found between neuroleptic dosage and rCBF. PMID- 2563934 TI - The training and utilization of surgical physician assistants. A retrospective study. AB - A community hospital's search for qualified surgical house staff in 1975 led to the development of a postgraduate residency program in surgery for physician assistants. Eleven years after its inception, the program's purpose and structure were reviewed, and its alumni, goals, and contributions were evaluated. A 1987 alumni survey provided data to assess the value of residency training to current employment and job satisfaction. PMID- 2563935 TI - Effect of beta-adrenergic blockade on the growth rate of abdominal aortic aneurysms. PMID- 2563936 TI - Familial clusters in human T-cell lymphotropic virus type I-associated myelopathy. PMID- 2563937 TI - Protirelin (thyrotropin-releasing hormone) in amyotrophic lateral sclerosis. The role of androgens. AB - Protirelin (thyrotropin-releasing hormone) appears to be a neuromodulator in the extrahypothalamic nervous system and has been suggested as an adjunct in the treatment of amyotrophic lateral sclerosis (ALS). Clinical studies have been divided on the efficacy of protirelin (TRH) despite strong experimental findings that are consistent with a role for the peptide in ALS. Recent findings provide evidence of a gender-related specificity in the ability of protirelin to potentiate the monosynaptic reflex. While castration in male neonatal rats lowered the sensitivity to protirelin, testosterone treatment restored that sensitivity. An examination of the clinical studies reveals a failure either to identify patients' sex or to separate the results on the basis of sex. These findings provide convincing evidence for the potential efficacy of protirelin in ALS if the patient's sex and underlying hormonal status are taken into account. PMID- 2563938 TI - Simultaneous presentation of parathyroid, thyroid and parotid tumours 44 years after neck irradiation. AB - A case of three tumours presenting 44 years following external irradiation to the neck is reported. Whilst thyroid, parotid and parathyroid tumours are all described in this situation, the simultaneous presentation of all three has not been documented previously. PMID- 2563939 TI - Involvement of D-amino acid oxidase in elimination of free D-amino acids in mice. AB - The physiological role of D-amino acid oxidase was investigated by using mutant ddY/DAO- mice lacking the enzyme. Free D-amino acid concentrations in the mutant mice were significantly higher than those of control ddY/DAO+ mice in kidney, liver, lung, heart, brain, erythrocytes, serum and urine. The results suggest that the enzyme is involved in the catabolism of free D-amino acids in the body, and that free D-amino acids are also excreted into urine. PMID- 2563940 TI - Steroid hormones inhibit binding of Vinca alkaloid to multidrug resistance related P-glycoprotein. AB - Multidrug-resistant cells are characterized by the presence of P-glycoprotein on the plasma membrane, which binds and probably transports antitumor agents outside the cells. P-glycoprotein is also present in various normal tissues such as the adrenal gland. To investigate the physiological function of P-glycoprotein, we examined possible endogenous materials which inhibit the binding of vincristine to the resistant cell membrane. The binding was inhibited by steroid hormones, most efficiently by progesterone. Progesterone also reduced the photoaffinity labeling of P-glycoprotein by a photoactive analogue of vindesine. These results suggest that P-glycoprotein in the adrenal gland could have a role in the secretion of steroid hormones. PMID- 2563941 TI - Guanosine 3':5'cyclic monophosphate and activators of guanylate cyclase inhibit secretagogue-induced corticotropin release by rat anterior pituitary cells. AB - The secretion of corticotropin by perfused rat anterior pituitary cell columns was studied. Forty-one residue corticotropin releasing factor, vasopressin and high extracellular KC1 all stimulated the secretion of corticotropin. The hormonal response to corticotropin-releasing factor (10(-10) mol/l), vasopressin (10(-9) mol/l) as well as KC1 (48 mmol/l) was reduced by membrane permeant analogs of cGMP, such as 8-BrcGMP and dibutyryl-cGMP. The 8-BrcGMP analog (10(-5) mol/l) inhibited corticotropin release in response to corticotropin-releasing factor by 30%, that to vasopressin by 70%, and that to KCl by 50%. Atriopeptin1 28 (10(-8) and 10(-7) mol/l), a peptide known to activate membrane-bound guanylate cyclase in the anterior pituitary gland, decreased the release of corticotropin induced by vasopressin to about 30% of control. Similarly, activators of soluble guanylate cyclase, such as glyceryltrinitrate and sodium nitroprusside (10(-5) mol/l) inhibited vasopressin-stimulated corticotropin release by 60%. In conclusion, the data show that purported activators of particulate and soluble guanylate cyclase, as well as derivatives of cGMP itself are strong inhibitors of secretagogue-induced corticotropin release by corticotroph cells of the anterior pituitary gland. PMID- 2563943 TI - A role of carbohydrate-carbohydrate interaction in the process of specific cell recognition during embryogenesis and organogenesis: a preliminary note. AB - A possible role of cell surface glycoconjugates in cell recognition has been envisioned based on recognition of carbohydrates by cell surface proteins such as endogenous lectins, glycosyltransferases, and hydrolases (refs. 18-22 in text). A new possibility that a specific carbohydrate at the cell surface could be recognized by the same or similar carbohydrate on the counterpart cell surface is now suggested by specific interaction between Lex and Lex, but not between Lex and sialylated or non-substituted type 2 chain. A new hypothesis is hereby proposed for carbohydrate-carbohydrate interactions as recognition signals during embryogenesis and organogenesis. PMID- 2563942 TI - Metabolite transport in rat kidney mitochondria: ornithine/phosphate translocator. AB - Ornithine uptake by rat kidney mitochondria is here first shown by monitoring the reduction of the intramitochondrial pyridine nucleotides which occurs as a result of metabolism of imported ornithine via ornithine aminotransferase and 1 pyrroline-carboxylate dehydrogenase. Ornithine uptake shows saturation features (Km and Vmax values, measured at 20 degrees C and at pH 7.20, were found to be about 0.85 mM and 23 nmoles/min x mg protein, respectively) and proves to be inhibited by D-ornithine, inorganic phosphate, praseodimium chloride and mersalyl. Neither malate nor glutamate, but phosphate was found to exchange with ornithine. Phosphate efflux caused by externally added ornithine was shown both as revealed by a c colorimetric assay and as continuously monitored by measuring extramitochondrial reduction of NAD+ in the presence of glyceraldehyde-3 phosphate, glyceraldehyde-3-phosphate dehydrogenase, ADP and 3-phosphoglycerate kinase. The role of ornithine carrier in kidney metabolism will also be discussed. PMID- 2563944 TI - Beta-blockers and hypertension. AB - While there is general agreement on the natural history, pathology, and pathophysiology of hypertension, there continues to be controversy over the selection of specific antihypertensive agents. All antihypertensive agents will, by definition, lower blood pressure, and factors beyond side effects and other difficulties associated with therapy form the basis of selecting specific agents. One of these factors is the effect of a given drug on core organ function. Propranolol was the first beta-adrenoceptor-blocking agent introduced for the treatment of hypertension. Initiation of therapy with propranolol may result in a decline in blood pressure more at the expense of cardiac function due to a concomitant rise in total peripheral resistance. Furthermore, propranolol may result in a decline in both glomerular filtration rate (GFR) and renal blood flow (RBF). In contrast, cardioselective beta-blockers or those with intrinsic sympathomimetic activity may not adversely affect renal function. It had been predicted that nadolol, a noncardioselective beta-blocker without intrinsic sympathomimetic activity, should result in decreased renal function. In contrast, observations demonstrated a preservation or improvement in both RBF and GFR, suggesting the presence of an alternative effective mechanism. Recent additions to the beta-adrenolytic group of antihypertensive agents include drugs with concurrent beta-blockade and vasodilation. This vasodilatation may be achieved through agonist properties resulting in lesser increases in vasomotor tone and smaller, if any, decreases in cardiac output. Alternatively, vasodilation may be achieved by concomitant alpha-adrenoceptor blockade, such as with labetalol. This agent preserves GFR and RBF during therapy of hypertension, in patients with normal as well as diminished renal function and hypertension.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563945 TI - Angiotensin-converting enzyme inhibitors in the treatment of mild to moderate essential hypertension. AB - The rationale for the use of new drugs in the treatment of mild to moderate hypertension is based on the knowledge that these drugs can both achieve the same goals as old drugs and offer additional advantages. Available data indicate that angiotensin-converting enzyme inhibitors (ACEIs) are as effective in reducing blood pressure as thiazide diuretics and beta-blockers, and that they maintain their effect without the development of tolerance. Therefore they may be expected to achieve at least the same prevention of target organ damage (heart failure, cerebral strokes, and arterial nephrosclerosis) as achieved by previous drugs and also to be effective when target organs have already been damaged. Moreover ACEIs seem to offer two possible advantages: cardioprotection and a favorable influence on the quality of life. Many experimental but indeed few clinical data suggest that these drugs can exert primary cardioprotection and that they might exert a beneficial therapeutic effect in hypertensives with coronary heart disease. Owing to the quality of their hypotensive effect, to the lack of detrimental effects on physical, psychical, and sexual activity, and to the positive influence on the sense of well-being, ACEIs seem to be better tolerated than previous drugs. However, although available data suggest that ACEIs can be considered a further advance in the treatment of hypertension, more extensive information is needed to confirm these promising results. PMID- 2563946 TI - Critical comments on recent literature. Age and the response to antihypertensive drugs. PMID- 2563947 TI - Population frequencies of tyrosine hydroxylase restriction fragment length polymorphisms in bipolar affective disorder. PMID- 2563948 TI - Cutis verticis gyrata associated with chronic schizophrenia in Chinese. PMID- 2563949 TI - One form of inclusion body beta-thalassemia is due to a GAA----TAA mutation at codon 121 of the beta chain. PMID- 2563950 TI - Studies on gamma-glutamyl transpeptidase in rodents exposed to benomyl. PMID- 2563951 TI - Coated mesalazine (5-aminosalicylic acid) versus sulphasalazine in the treatment of active ulcerative colitis: a randomised trial. AB - OBJECTIVE: To assess the safety and efficacy of a preparation of mesalazine (5 aminosalicylic acid) coated with a pH dependent resin (Eudragit L) as compared with sulphasalazine in patients with active mild to moderate ulcerative colitis. DESIGN: Eight week randomised double blind parallel group study. SETTING: Forty six gastroenterology outpatient clinics in seven countries. PATIENTS: Two hundred and twenty patients aged 18-70 who met the following criteria: clinical activity index greater than or equal to 6 and endoscopic index greater than or equal to 4; no concomitant treatment for ulcerative colitis; no hypersensitivity to salicylates or sulphonamides. Of the 164 patients eligible for efficacy analysis, 87 received the coated preparation of mesalazine and 77 sulphasalazine. Most of the remaining patients (28 in each group) were ineligible for the efficacy analysis because of treatment with steroid enemas. All pretrial characteristics were comparable in the two treatment groups. INTERVENTIONS: Coated mesalazine (Mesasal) 1.5 g daily or sulphasalazine 3.0 g daily for eight weeks. Compliance monitored by pill counts. END POINT: Clinical and endoscopic remission. MEASUREMENTS AND MAIN RESULTS: Clinical activity measured by daily diary cards, assessment by investigators, and laboratory findings. Endoscopic evaluation at week 8. After four weeks 50 of 70 patients (71%) taking coated mesalazine and 38 of 58 (66%) taking sulphasalazine had achieved remission of their disease by eight weeks remission rates were 74% (37/50 patients) and 81% (35/43) in the two treatment groups respectively. Endoscopic remission at eight weeks was recorded in 20 of 41 patients (49%) taking coated mesalazine and 18 of 38 (47%) taking sulphasalazine. There was a higher incidence of adverse events among patients taking sulphasalazine (25/105; 24%) than among those taking coated mesalazine (16/115; 14%). CONCLUSION: Mesalazine coated with Eudragit L is a safe, logical alternative to sulphasalazine. PMID- 2563952 TI - Treatment of itching in atopic eczema with antihistamines with a low sedative profile. PMID- 2563954 TI - Effect of antianginal medications on the prognostic value of exercise thallium scintigraphy. AB - Whether administration of antianginal medications at the time of exercise thallium scintigraphy reduces the prognostic value of this test was retrospectively examined using two year follow-up of 201 patients. Sensitivity, specificity, positive and negative predictive values and accuracy of five test outcomes for prediction of coronary events (unstable angina, myocardial infarction, cardiac death) were compared between groups of patients either taking or not taking antianginal medications. Specificity and negative predictive value of exercise and redistribution thallium scores for prediction of coronary events were greater in patients not taking antianginal medication (P less than 0.05). It was concluded that normal exercise thallium scintigraphy affords greater assurance against future coronary events in patients tested while not taking antianginal medications. PMID- 2563953 TI - Study of the in vitro bioactivation of albendazole in human liver microsomes and hepatoma cell lines. AB - The metabolism of albendazole (ABZ), a benzimidazole anthelminthic, was studied in either microsomal preparations of human liver biopsies or cultured human hepatoma cell lines. Metabolites were analyzed by HPLC. Our data show that microsomes from human biopsies and two human cell lines, HepG2 and Hep3B, oxidize the drug to the sulfoxide very efficiently, whereas the third cell line tested, SK-HEP-1, does not. Both cytochrome P-450 dependent monooxygenases and flavin containing monooxygenases appear to be involved in human ABZ metabolism. Using the cell line displaying the highest ABZ-metabolizing activity, HepG2, the cytotoxic and the inducing effects of the parent drug ABZ and of two primary metabolites, the sulfoxide and the sulfone were studied. These three chemicals provoked a rise in mitotic index resulting from cell division blockage at the prophase or at the metaphase (ABZ metabolites) stage, and ABZ was more cytotoxic than its metabolites. With regard to enzyme-inducing effects, our data clearly demonstrate that the sulfoxide and, to a lesser degree, the sulfone are potent inducers of some drug metabolizing enzymes (i.e., cytochrome P-488 dependent monooxygenases and UDP glucuronyltransferase), whereas ABZ fails to increase and even slightly decreases these enzymatic activities. In conclusion, the HepG2 human hepatoma cell line appears to be suitable for the study of many parameters of metabolism and action of ABZ and other structurally related compounds in humans. PMID- 2563955 TI - Hemodynamic and adrenergic responses of bevantolol and propranolol in hypertensives. AB - The hemodynamic and neurohumoral responses of 21 thiazide-resistant hypertensives receiving sequential chronic therapy with propranolol and bevantolol, a new cardioselective beta-1 blocker, were studied and compared with their responses to placebo. The objective was to determine to what extent decreased circulation levels of catecholamines and renin activity contributed to the hypotensive action of bevantolol and whether it demonstrated a significant sparing effect on vascular resistance. Both propranolol and bevantolol lowered supine and erect blood pressures to a comparable extent but the response of diastolic pressure to upright posture was maintained. Resting heart rates were lowered and postural tachycardia was attenuated. Propranolol induced a greater decrease in forearm blood flow and greater increase in vascular resistance than bevantolol. Both drug therapies were associated with lowered plasma concentrations of noradrenaline and adrenaline, while the decrease in noradrenaline levels was linearly related to the fall of mean arterial pressure for both drugs. Plasma renin activity was lowered only to a marginal extent by either drug but aldosterone concentrations were significantly reduced to a comparable extent by both drugs. The results suggest that a negative chronotropic action on the heart and an overall reduction in sympathetic nervous tone both contribute to the hypotensive effects of bevantolol and propranolol, but reduction of plasma renin activity may be of lesser importance. Bevantolol demonstrated a significant vascular sparing effect in this patient group compared with propranolol. PMID- 2563956 TI - Elevation of sialyl stage-specific mouse embryonic antigen levels in pleural effusion in patients with adenocarcinoma of the lung. AB - Sialyl stage-specific mouse embryonic antigen (SSEA-1) levels were measured in pleural effusions obtained from patients with lung cancer and benign pulmonary disease, using a solid-phase immunoradiometric sandwich assay. The mean (+/- SEM) levels (unit/ml) of pleural fluid sialyl SSEA-1 were 3620 +/- 1419 in adenocarcinoma (n = 25), 123 +/- 30 in nonadenocarcinoma (n = 13) and 95 +/- 19 in benign pulmonary disease (n = 13), respectively. The positive rate was 64% in adenocarcinoma, 7.7% in nonadenocarcinoma, and 0% in benign pulmonary disease, respectively, when a cutoff level was defined as the mean + 3 SD value (300 unit/ml) based on pleural fluid sialyl SSEA-1 levels in benign pulmonary disease. There was a significant positive correlation between pleural fluid levels of sialyl SSEA-1 and those of carcinoembryonic antigen in adenocarcinoma patients (r = 0.8246, P less than 0.01). Pleural fluid sialyl SSEA-1 levels correlated with cytologic findings in adenocarcinoma patients. These observations suggest that sialyl SSEA-1 in pleural effusion is a useful marker to discriminate malignant from nonmalignant and adenocarcinoma from nonadenocarcinoma of the lung. PMID- 2563957 TI - Treatment of mouse thymus cells with phosphatidylinositol-specific phospholipase C preferentially abrogates their reactivity with autoantibodies to the Thy-1 antigen. AB - Thy-1 molecules have been shown to become anchored to the membrane of thymocytes and of T-cells via a phosphatidylinositol link. In the present study the intensity of immunofluorescent staining of mouse thymus cells by monoclonal xenoantibodies and alloantibodies specific for the Thy-1.2 determinant was significantly reduced following exposure of the cells to phosphatidylinositol specific phospholipase C (PI-PLC). The majority of the PI-PLC-treated thymus cells retained, however, some reactivity with Thy-1.2 antibodies. In contrast, the immunofluorescent staining of thymus cells with monoclonal autoantibodies to Thy-1 determinants (20-10-5 and C16-31) was completely abolished by PI-PLC treatment. These results suggest that whereas monoclonal autoantibodies to Thy-1 react preferentially with PI-PLC-sensitive Thy-1 molecules, monoclonal antibodies to the Thy-1.2 specificity react with all cell surface Thy-1 molecules, regardless of their sensitivity to PI-PLC treatment. PMID- 2563959 TI - Isolation and characterization of dipeptidyl aminopeptidase IV from human kidney cortex. AB - Intact dipeptidyl aminopeptidase IV (DAP IV) was solubilized by bromelain treatment from human kidney brush border plasma-membranes. Purification of DAP IV was performed by a 3-step method, applying lectin-affinity chromatography on WGA Sepharose, gel filtration and anion-exchange chromatography. DAP IV from human kidney cortex showed a pH optimum of 8.7 and was totally inhibited by 1 mmol/l Zn2+. Isolated DAP IV revealed a relative molecular mass of 250 kDa as determined by the native-PAGE method and of 220 kDa by the gel filtration method. Analytical isoelectric focussing of DAP IV revealed an isoelectric point of pH 5.3. Ultrastructural analysis of isolated DAP IV fractions, using the negative staining technique, disclosed the presence of numerous globular particles with an average diameter of 5 nm which correspond to the structural substrate of the purified protein. PMID- 2563958 TI - Clinical significance of a new isoform of serum alanine aminopeptidase; relationship with liver disease and alcohol consumption. AB - The potential clinical interest of a new isoform of alanine aminopeptidase (AAP) was examined using polyacrylamide gradient gel electrophoresis. This form, called F-AAP, is more proteolyzed than normal serum AAP. It appears to be associated with many liver diseases and is related to cytolysis, but also to elevated GGT levels. The F-AAP form was present in 52% of subjects treated with anticonvulsant drugs and in 64% of subjects consuming more than 44 g of alcohol per day. Analysis of F-AAP may be of interest in detecting effects of drugs and alcohol on the liver. PMID- 2563960 TI - Surgical pearls. AB - This article contains a potpurri of surgically related stratagems, alternative techniques, and philosophies. These surgical pearls have been selected for their simplicity, usefulness, and applicability. PMID- 2563961 TI - Treatment of an enteric fistula with somatostatin in a premature. AB - The authors present the case of an extremely premature baby affected by severe Necrotizing Enterocolitis (NEC) needing intestinal resection and ileostomy. An enteric fistula developed 3 days after ostomy closure. The baby was started on Total Parenteral Nutrition (TPN), and 7 days later the fistula output remained constant. Somatostatin (SM) was then given intravenously (3.0 micrograms/Kg/hr) and the fistula closed on the 3rd day of treatment. Since SM was introduced in 1986 as an adjunct treatment to TPN in enteric fistulas, the authors believe that theirs is the first report of a successful SM treatment in a premature. PMID- 2563962 TI - Pharmacokinetics of esmolol and ASL-8123 in renal failure. AB - The effect of renal function on the pharmacokinetics of esmolol, an ultra-short acting beta-adrenergic blocker, and its major metabolite, ASL-8123, was examined in six healthy control subjects, six patients maintained on hemodialysis, and six patients on continuous ambulatory peritoneal dialysis (CAPD). In addition, the impact of hemodialysis and CAPD on removal of esmolol and ASL-8123 was determined. Multiple blood, urine, and dialysate samples were collected during a 72-hour period and assayed for esmolol and ASL-8123 by HPLC. The pharmacokinetic disposition of esmolol was not significantly altered by renal failure. Mean (+/- SD) total body clearance for esmolol was 171.4 +/- 69.8, 249.8 +/- 176.3, and 265.3 +/- 143.1 ml/min/kg for the control, hemodialysis, and CAPD patients, respectively. Mean elimination half-life (t1/2) was 7.2 minutes in control subjects compared with 7.1 and 8.0 minutes for the hemodialysis and CAPD groups, respectively. The apparent volume of distribution of esmolol did not differ significantly among the three groups. ASL-8123 was shown to accumulate in patients with renal failure, as evidenced by a mean maximum blood concentration of 42.8 +/- 12.2 micrograms/ml in the control group compared with 76.1 +/- 23.9 and 87.1 +/- 20.4 micrograms/ml in the hemodialysis and CAPD groups, respectively (p less than 0.05). The elimination t1/2 of ASL-8123 was prolonged in patients with renal failure, averaging more than 42 hours compared with only 4 hours in the control subjects. Approximately 20% of the esmolol dose as ASL-8123, was removed by either hemodialysis or CAPD, contributing minimally to the elimination of the drug.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563964 TI - Somatostatin in gastroenterology. American Gastroenterologic Association, plenary session papers. Chicago, Illinois, May 1987. PMID- 2563963 TI - Pancreatic enzyme replacement therapy. Importance of gastric acid secretion, H2 antagonists, and enteric coating. AB - The relative efficacy of three commercial pancreatic enzyme supplements in improving fat absorption was studied using the [14C]triolein breath test in 12 patients with chronic pancreatitis. Two of the supplements were enteric coated. The one nonenteric coated product was studied twice: with and without ranitidine coadministration. Doses complied with the manufacturers recommendations. Baseline studies included pentagastrin-stimulated gastric acids, 72-hr fecal fat excretion, and [14C]triolein absorption while not on supplementation. Acid outputs were variable (BAO: 0.3-4.1 meq/hr; MAO: 3.5-34.6 meq/hr). Three patients had mild steatorrhea (i.e., less than 10 g/day) and the remaining severe fat malabsorption (56.9 +/- 41.5 g/day). Although fat absorption was significantly improved by all three supplements, the nonenteric coated preparation was most effective (P less than 0.001). However, laboratory analysis demonstrated that lipase content was four times greater, ie, 17,000 IU/4 tablets. Pretreatment with ranitidine failed to further improve the absorption in patients given nonenteric supplements but was effective in those found to have high or normal acid outputs (P less than 0.001). Our results suggest that the recommended dosage of enteric coated preparations is insufficient for adult patients with severe chronic pancreatitis. Secondly, the marked variability of acid secretion in such patients possibly accounts for the variability of results obtained by others on the usefulness of coadministration of antacids and H2 antagonists. Routine measurement of gastric acid secretion status may help optimize the choice and form of pancreatic enzyme supplementation. PMID- 2563965 TI - Somatostatin in portal hypertension. AB - The effect of somatostatin on portal pressure is mediated by splanchnic arterial vasoconstriction which induces a reduction in portal blood flow and pressure. One of the most important characteristics of somatostatin is that its splanchnic effect is not accompanied by major systemic hemodynamic effects. Somatostatin has been used in several controlled trials to test its potential in controlling acute variceal bleeding. The results remain controversial. Different findings in existing clinical trials may derive in part from distinct protocols for somatostatin administration. Published trials suggest that somatostatin may be as effective as vasopressin in the acute management of variceal bleeding. However, since the efficacy of vasopressin has been questioned, a comparison of two potentially ineffective drugs cannot establish definitively the efficacy of somatostatin in controlling variceal bleeding. The most significant finding of the two published studies has been the lower incidence of minor and major complications with somatostatin when compared to vasopressin. Newer trials in progress may shed new light into the potential use of somatostatin for the treatment of variceal bleeding. PMID- 2563966 TI - Biochemistry and physiology of gastrointestinal somatostatin. AB - Somatostatin, a tetradecapeptide initially isolated from the ovine hypothalamus, is widely distributed throughout the gastrointestinal tract where it may act as a hormone, local chemical messenger, or neurotransmitter to elicit many physiological actions. Release of somatostatin from D cells in the gut is regulated by mechanisms that are both dependent on and independent of cAMP. In most cases somatostatin acts to inhibit the function of its target cells. It performs this action in part via pertussis-toxin-sensitive inhibitory guanine nucleotide-binding proteins that regulate adenylate cyclase activity. Other mechanisms may involve sites of action distal to intracellular second messenger systems. PMID- 2563967 TI - Transferrin and mitochondrial aspartate aminotransferase in young adult alcoholics. AB - Two recently proposed biochemical markers of alcoholism, namely, quantitation of plasma transferrin variant (Tf5.7) and the ratio of plasma mitochondrial aspartate aminotransferase (m-AspAT) to total AspAT (t-AspAT), were tested for their ability to detect young adult alcoholics. Another commonly used biochemical test, namely, activity of plasma gamma glutamyltransferase (GGT) was included as a comparison. Although mean values of GGT, TF5.7, total transferrin (Tftot), m AspAT and t-AspAT in alcoholics were significantly higher than those in controls, there were too many overlapping values in each test between alcoholics and controls to render any of these tests suitable as a marker for young adult alcoholics. Depending on cut-off limits, the sensitivity of each test ranged from 0-52% and the specificity ranged from 80-97%. Moreover, the m-AspAT/t-AspAT and Tf5.7/Tftot ratios were not significantly different between alcoholics and controls. A stepwise linear discriminant function analysis of all the variables resulted in a slight increase in classification sensitivity (66%) but a decrease in specificity (77%). The relatively short duration (mean = 5.6 years) of heavy alcohol intake and the time elapsed (mean = 5.8 days) since the alcoholics last consumed alcohol very likely contributed to the low sensitivity. Young adults might also be more resilient with regard to the damaging biochemical effects of ethanol. Abnormal biochemical values might reverse to normal values much more quickly in young adult alcoholics than in those who are older and have more years of alcohol abuse. PMID- 2563968 TI - Effect of ethanol on the maturation of the spontaneous transmitter release by regenerated nerve endings. AB - The cellular mechanisms involved in the theratogentic action of ethanol are not well known, but neuron outgrowth and synaptogenesis are regarded as the periods in which ethanol causes its major damage in the nervous system. The effects of chronic treatment with ethanol on the maturation of the spontaneous transmitter release by regenerated nerve endings in the rat were studied. The sciatic nerve was crushed and miniature end plate potentials (mepps) were recorded intracellularly in the re-innervated extensor digitorum longus muscle at different points in time after denervation; end plate potentials (epps) were also recorded. Two main effects were observed in the re-innervated muscles of ethanol treated rats: (1) the appearance of spontaneous and evoked transmitter release was delayed and (2) the subsequent increase in frequency of mepps is faster. The possible mechanisms involved in these effects are discussed. PMID- 2563969 TI - Narcotic analgesics, their detection and pain measurement in the horse: a review. AB - Narcotic analgesics produce pharmacological effects by interacting with specific opiate receptors. At least five major types of opiate receptors have been recognised. These include mu (morphine) and kappa (ethylketazocine) receptor types. Narcotic analgesics which interact with mu receptors produce locomotor and autonomic stimulation at doses that produce little or no analgesia. Therefore, use of these drugs as analgesics in equine medicine has not been very satisfactory. Theoretical considerations suggested that the role of kappa agonists in equine analgesia be investigated. Using a pure kappa agonist, U-50, 488H, good analgesia was produced in the horse with little or no locomotor stimulation or autonomic effects. These data suggest that kappa agonists may be superior analgesics for clinical use in the horse. On the other hand, the locomotor stimulant effects of mu agonist analgesics enable their use as illegal medications. Specifically, these agents produce a good running response, signs of central nervous stimulation and analgesia, all potentially useful effects in a racehorse. Regulatory control of most narcotic analgesics can be obtained by high performance thin layer chromatographic screening. However, effective screening for the fentanyls and small doses of etorphine can only be achieved by use of immunoassay. PMID- 2563970 TI - Iodine-131 MIBG scintigraphy in small cell lung cancer. AB - There is a well documented relationship between small cell carcinoma of the lung and the amine precursor uptake and decarboxylation system of endocrine cells (APUD). We attempted to exploit this association by employing the unique radiopharmaceutical, 131I-MIBG, which is recognized and taken up by the APUD system to monitor disease activity in patients with small cell carcinoma of the lung. A total of eight patients with biopsy proven, metastatic small cell carcinoma of the lung were studied. 131I-MIBG was synthesized in our laboratory by reacting metaiodobenzylamine hydrochloride with cyanamide with subsequent solid phase radioiodination. A dose of 0.5 mCi radiopharmaceutical was injected and images obtained on a large field of view gamma camera with a high energy parallel hole collimator at 2, 24, and either 48 or 72 h. Images were compared with known focal areas of metastatic disease demonstrable on computed tomographic scan, chest roentgenogram or bone scan. We were unable to detect reproducible correlations between the images produced by conventional radiographic techniques and the images produced by our radiopharmaceutical. We conclude that this agent will probably not be useful for localization of metastatic small cell lung carcinoma. PMID- 2563971 TI - Different mechanisms account for the suppression of interleukin 2 production and the suppression of interleukin 2 receptor expression in Trypanosoma brucei infected mice. AB - Lymph node cell populations derived from Trypanosoma brucei-infected mice failed to produce interleukin 2 (IL 2) in response to a potent mitogenic trigger and suppress the potential of normal lymph node cells to secrete IL 2 in co-culture assays. This suppression is promptly restored by the addition of indomethacin, which blocks prostaglandin synthesis, but is not markedly affected by the addition of catalase, which degrades H2O2. The suppression of the IL 2 receptor expression, on the other hand, is not restored by the addition of indomethacin, nor by the simultaneous supply of both indomethacin and catalase. This discrepancy is not caused by an extreme susceptibility of the receptor expression to low prostaglandin (PG) concentrations, but rather by the presence of suppressive cells that operate through a PG-independent mechanism. This suppressive mechanism accounts for the loss of the IL 2 receptors on both the Ly 2+ and the L3T4+ T cell compartment. The indomethacin-treated co-cultures, which manifest a normal IL 2 production but lack the IL 2 receptors, manifest an impaired DNA synthesis and contain a decreased number of T cell blasts. PMID- 2563972 TI - Evidences for protein kinase C. Activation in T lymphocytes by stimulation of either the CD2 or CD3 antigens. AB - Phosphorylation of protein kinase C (PKC) substrates in T lymphocytes was analyzed after stimulation with specific pairs of anti-CD2 monoclonal antibodies (mAb) or an anti-CD3 mAb. The results show that the appropriate stimulation of both CD2 or CD3 antigens results in phosphorylation of a 80-kDa putative PKC substrate and that this phosphorylation event is sensitive to a PKC inhibitor, sphinganine. CD2- and CD3-dependent phosphorylation was found to be strongly dependent on an extensive cross-linking of surface antigens. The biological importance of cross-linking of CD2 and CD3 was also evident for other biological responses such as interleukin 2 production and induction of an autocrine growth response. Finally, we also present evidence for interaction between the CD2 and CD3 signal transducing pathways. PMID- 2563973 TI - Restriction fragment analysis of V preB and lambda 5 within the genus Mus. AB - DNA from a panel of inbred strains of mice and colony bred mice, isolated from different geographical locations, was hybridized to mouse V preB and lambda 5 probes under stringent conditions, indicating sequence similarities greater than 80%. The probe for lambda 5 detects one gene and the probe for V preB detects two genes (V preB1 and V preB2) in the inbred strains of mice examined under the stringency used. No restriction endonuclease fragment length polymorphisms (RFLP) were detected with the V preB and lambda 5 DNA probes among the inbred strains of mice using Bam HI and Hind III. Very few RFLP were detected among Mus musculus subspecies, and the intensity of the hybridization did not differ significantly with either DNA probe. The number of RFLP increased slightly when different species and subgenera were examined, and the intensity of the hybridization signal began to decrease in samples from the different subgenera, suggesting a slight decrease in sequence similarity for both V preB genes with increased time of divergence. Fewer RFLP were detected with the lambda 5 DNA probe. DNA from 11 different Mus species representing 4 subgenera, genetically isolated from laboratory mice for approximately 1-12 million years, continued to hybridize under high stringency conditions using both DNA probes. A comigrating lambda 5 and V preB restriction endonuclease fragment was detected in most of the samples examined, suggesting the close physical linkage of V preB1 and lambda 5 is maintained within the genus Mus. These results suggest that V preB1, V preB2 and lambda 5 have been present for over 12 million years. PMID- 2563974 TI - Intrathecal bradykinin acts presynaptically on spinal noradrenergic terminals to produce antinociception in the rat. AB - In the awake restrained rat the intrathecal (i.th.) administration of 8.1 pmol 8.1 nmol of bradykinin (BK) and kallidin (KD) enhanced the reaction time (RT) to a noxious radiant heat stimulus in a dose-dependent manner. The fragments BK-(1 8) and BK-(1-7) were active only at doses higher than 10 nmol and the following rank order of potency was observed: BK greater than KD much greater than BK-(1-8) greater than BK-(1-7). The increment of tail-flick latency was greatest at 1 (BK) or 6 (KD) min and the RT returned to basal levels within 15 min post administration. The effect of BK (81 pmol) was unaffected by the prior i.th. administration of propranolol and naloxone but was significantly potentiated by prazosin (P less than 0.05). In contrast, the response to BK was significantly blocked (P less than 0.001) by phentolamine, idazoxan and yohimbine as well as by treatment with 6-hydroxydopamine (6-OHDA) at a dose of 20 micrograms (i.th.) 1 week earlier. The latter pretreatment reduced the antinociceptive effect of i.th. tyramine (7 mumol) and potentiated that to noradrenaline (NA) (0.6 nmol) (P less than 0.01) while it preserved both the antinociceptive effect of neurokinin B (8 nmol) and the hyperalgesic effect of substance P (6.5 nmol). A biochemical analysis revealed that 6-OHDA treatment reduced the NA content in the lumbar spinal cord by 60% without affecting the levels of serotonin, dopamine, adrenaline or their main metabolites. There were also significant reductions in NA content in cervical (44%) and thoracic (55%) spinal cord. Pretreatment with 6 OHDA for a longer survival period (2 weeks) caused a further decrease of NA in the lumbar spinal cord (88%); however, the serotonin and dopamine levels were reduced in all regions examined. These results suggest that BK (kinins) may inhibit spinal nociceptive sensory transmission and produce analgesia by acting presynaptically on terminals of bulbospinal NA-containing fibers. PMID- 2563975 TI - Biochemical analysis of young rats homozygous for the cataract mutation cat. AB - Cataractogenesis was studied in young rats homozygous for the radiation-induced recessive cataract mutation cat. Homozygous cat/cat rats have reduced body weight (about two-thirds of the wild type) when 3 weeks old. The litter size is also diminished to about two-thirds of the wild type. For lens-specific parameters, as compared with homozygous wild type, the wet weight of the cataractous lenses is reduced, although the concentration of water-soluble lens proteins per wet weight is the same. No major alterations could be detected in the pattern of the water soluble lens proteins separated by isoelectric focusing or gel electrophoresis run with or without mercaptoethanol. Additionally, no statistically significant alterations could be detected in the biochemical parameters of the lens used as indicators for osmotic stress (water content of the lens and the Na+-K+-dependent ATPase), for the energy state (ATP) and for the redox state (oxidized glutathione and superoxide dismutase). In contrast, the activity of transglutaminase is significantly enhanced in lenses as well as in the liver of young cat-rats, which might be understood as a biochemical marker for alterations in the developmental program. Cataractogenesis in the cat-rat is, therefore, suggested to be part of a syndrome including dwarfism and reduced litter size. PMID- 2563976 TI - Expression of the mutant gene for L-gulono-gamma-lactone oxidase in scurvy-prone rats. AB - A mutant strain of Wistar rats with L-gulono-gamma-lactone oxidase deficiency has recently been established. To investigate this deficiency by DNA and RNA blot hybridization analyses, a fragment of a previously cloned cDNA encoding rat L gulono-gamma-lactone oxidase was used as a probe. When genomic DNA of the mutant rat was digested with several restriction enzymes, the probe hybridized to fragments of the same sizes as those produced from DNA of normal rats. Poly(A)+RNA from the liver of the mutant rat was found to contain an L-gulono gamma-lactone oxidase-specific mRNA of a normal size at a comparable level to that of normal rats. An in vitro translation experiment revealed that the mRNA programmed the synthesis of an enzyme protein which had the same molecular weight as that of the translational product of the normal mRNA, although the amount synthesized was markedly reduced as compared with that synthesized with the normal mRNA. In accordance with this observation, a very low but definite degree of L-gulono-gamma-lactone oxidase activity was detected in the microsomes of the mutant rat by a newly developed, highly sensitive method. PMID- 2563978 TI - Presynaptic effects of spider toxins: increase of high affinity uptake in the arthropod peripheral glutamatergic system. AB - In contrast to the reported effects of polyamines on the high affinity neurotransmitter uptake, two polyamine-like spider toxins significantly increase the high affinity uptake of glutamate as demonstrated with high resolution autoradiography. The effects of both spider toxins were compared to those of a polyamine toxin from the wasp Philanthus triangulum, which is known to inhibit the high affinity glutamate uptake. PMID- 2563977 TI - Immunohistochemical localization of glutamine synthetase in human liver. AB - Glutamine synthetase (GS) of human liver was recognized with a polyclonal antibody to pig brain GS, but failed to stain with an antibody against rat liver GS. Using the latter antibody GS of human liver was shown to be localized within small rings of 1 to 3 hepatocytes surrounding the terminal hepatic venules. This pattern was analogous to that seen in rat and mouse liver. PMID- 2563980 TI - The hypothalamic somatostatinergic pathways mediate feeding behavior in the rat. AB - The level of somatostatin in the hypothalamus was higher in satiated rats than in hungry rats. Elevating hypothalamic somatostatin levels by administering somatostatin into the hypothalamus produced a decrease in food intake, whereas lowering hypothalamic somatostatin levels by administering cysteamine into the peritoneal cavity produced an increase in food intake in rats. PMID- 2563979 TI - Late inhibitory postsynaptic potentials in rat prefrontal cortex may be mediated by GABAB receptors. AB - The GABAB antagonist phaclofen blocked the postsynaptic hyperpolarization induced by the GABAB agonist baclofen during intracellular recordings in rat cortical cells. This effect appears to be selective since responses to GABAA agonists (muscimol, THIP), GABA, 5-HT and L-glutamate were unaffected. Phaclofen also blocked synaptically evoked late inhibitory postsynaptic potentials (late IPSP). These results suggest that the late IPSPs in cortical neurons are mediated by GABA acting on GABAB receptors. PMID- 2563981 TI - The new genetics: molecular technology and reproductive biology. PMID- 2563982 TI - Variations of somatostatin-like immunoreactivity in the circumoesophageal ganglia, the hepatopancreas, the mantle edge, and the hemolymph of shell repairing snails (Helix aspersa). AB - Immunocytology and radioimmunoassays demonstrate the presence of immunoreactive somatostatin-like (IR-SOM-LI) material in the tissues of young or adult snails Helix aspersa bred under short or long day in controlled artificial conditions. Neurosecretory cells in the circumoesophageal ganglia and in the digestive gland, small fibers in the hepatopancreas, and small granules in the mantle epithelial cells are immunoreactive toward antisomatostatin. In all experimented animals shell fractures induce variations of the IR-SOM-LI concentrations in all assayed tissues whatever the lighting conditions were underwent. These findings support the hypothesis that in Helix one or more substances related to the vertebrate tetradecapeptide is (are) involved in the repair processes but that the storage and the metabolism may be different during the biological cycle of the snails. These results are compared with those previously published using different gastropods and different methods. PMID- 2563984 TI - Localization of ammonia-metabolizing enzymes in human liver: ontogenesis of heterogeneity. AB - Immunohistochemical analysis of human liver (8 to 94 years) shows a compartmentation of ammonia-metabolizing enzymes across the acinus. The highest concentration of carbamoylphosphate synthetase (ammonia) is found in the parenchymal cells around the terminal portal venules. Glutamine synthetase is found in a small pericentral compartment two to three cells thick. In contrast to observations in rat liver, in human liver a well-recognizable intermediate zone can be distinguished in which neither enzyme can be detected. This intermediate zone is not yet established at the age of 8 years but can be recognized in livers from 25 years onward. Carbamoylphosphate synthetase can already be detected in the liver of human fetuses at 5 weeks of development. The enzyme distribution reveals a random heterogeneity among the hepatocytes, suggesting that not all hepatocytes start to accumulate carbamoylphosphate synthetase at the same time. From 9 weeks of development onward, the enzyme becomes homogeneously distributed throughout the liver parenchyma until at least 2 days after birth. Glutamine synthetase cannot be detected during this period. In addition, the definitive architecture of the acinus is not yet completed at birth. These results therefore support the idea that in human liver, metabolic zonation with respect to NH3 metabolism exists as it does in rat liver. Furthermore, the data show that this functional compartmentation becomes established concomitant with the development of the acinar architecture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563983 TI - Early biochemical responses of the small intestine of coeliac patients to wheat gluten. AB - The pathogenesis of coeliac disease has been investigated by studying the response of small intestinal hydrolases in patients with coeliac disease subject to gluten challenge. Small intestinal biopsies taken before and two and a half hours after a gluten challenge in five patients with coeliac disease who had been maintained on a gluten free diet were examined by a combination of electron and light microscopy, organ culture, pulse chase biosynthetic labelling, SDS-PAGE and autoradiography. Before the challenge, the small intestinal biopsies showed nearly normal morphology. Two and a half hours after the challenge there was deterioration in villus architecture, distortion of microvillus structure, disorganisation of the intermicrovillus pit region, an increase in lysosome like bodies in the apical cytoplasm of the luminal enterocytes and pronounced hypertrophy of the rough endoplasmic reticulum of these cells. SDS-PAGE of small intestinal biopsies from four treated coeliac patients before gluten challenge revealed normal microvillus membrane and hydrolase composition. There was a generalised reduction but no specific alteration in the pattern of polypeptide synthesis in the mucosa of the small intestine in these subjects two and a half hours after the gluten challenge. These results suggest that the generalised reduction in small intestinal brush border enzymes in coeliac patients is not the primary pathogenetic mechanism and represents a secondary effect. PMID- 2563985 TI - Beta-blockade prevents recurrent gastrointestinal bleeding in well-compensated patients with alcoholic cirrhosis: a multicenter randomized controlled trial. AB - To assess the efficacy of beta-blockers in preventing rebleeding in selected cirrhotic patients and to compare the tolerance, safety and patient compliance of a selective and a nonselective beta-blocker, 94 patients were randomly assigned to propranolol (32 patients), atenolol (32 patients) or placebo (30 patients). Randomization was made at least 15 days after the bleeding episode. Propranolol was given orally at increasing doses until the resting pulse rate was reduced by approximately 25%. Atenolol was given at a fixed dose of 100 mg per day. Patients were followed up for a mean of 357 days. Rebleeding occurred in 14 patients in the placebo group, 10 in the atenolol group and eight in the propranolol group. The incidence of rebleeding was significantly lower in patients receiving propranolol than in those on placebo (propranolol vs. placebo: p = 0.01, logrank test). Atenolol was less effective than propranolol (atenolol vs. placebo: p = 0.065, logrank test). Bleeding-free survival was better for patients on active drugs than for those on placebo (propranolol vs. placebo = p = 0.01, atenolol vs. placebo: p = 0.05, logrank test). Retrospective analysis revealed that, whatever the type of treatment, abstinence from alcohol was crucial in preventing rebleeding. We conclude that beta-blocker treatment is effective in preventing rebleeding from esophageal varices in carefully selected alcoholic cirrhotic patients who survive at least 2 weeks after acute variceal hemorrhage and stop drinking. PMID- 2563986 TI - Enzyme profile of rat bile ductular epithelial cells in reference to the resistance phenotype in hepatocarcinogenesis. AB - An extensive bile ductular cell hyperplasia with the formation of well differentiated bile ductules is the most prominent feature of rat liver at 6 to 15 weeks after bile duct ligation. We have improved our previous cell isolation procedure and are now routinely able to obtain from such livers high yields of viable bile ductular epithelial cells. These cells were characterized with respect to their specific activities of gamma-glutamyl transpeptidase and beta glucuronidase and of select Phase I and Phase II enzymes of biotransformation. At the time of their isolation, only a very small number of the bile ductular epithelial cells were observed to be in DNA synthesis. In addition, in histological sections prepared from intact hyperplastic bile ductular tissue isolates, only the bile ductular epithelial cells exhibited histochemical staining for gamma-glutamyl transpeptidase activity. Typically, greater than 95% of the cells isolated from this tissue were also found to be histochemically positive for gamma-glutamyl transpeptidase activity, and no hepatocytes were seen contaminating this cell population. Biochemically, the isolated bile ductular cells exhibited a gamma-glutamyl transpeptidase specific activity that was 100 times higher than that of hepatocytes isolated at the same time from the bile duct-ligated rats and more than 300 times higher than the specific activity of the enzyme of freshly isolated normal rat hepatocytes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2563987 TI - Leucocyte common antigen in the differential diagnosis of Hodgkin's disease. AB - The utility of immunohistochemical staining for leukocyte common antigen (LCA) in the differential diagnosis of Hodgkin's disease was studied in a series of 42 cases of Hodgkin's and non-Hodgkin's lymphoma and compared with Leu M-1, a proposed marker for Reed-Sternberg cells. LCA staining of neoplastic cells was absent in 23 of 24 cases of Hodgkin's disease but present in 16 of 18 cases of non-Hodgkin's lymphoma including eight of 10 cases with pleomorphic Reed Sternberg-like cells. Leu M-1, in contrast was present in 14 of 24 cases of Hodgkin's disease, but also in five of 18 cases of non-Hodgkin's lymphoma including five of 10 cases with pleomorphic Reed-Sternberg-like cells. Immunohistochemical staining for LCA appears useful in the differential diagnosis of Hodgkin's disease and appears to be a better discriminant than Leu M-1. PMID- 2563988 TI - Study of restriction fragment length polymorphisms at the human phenylalanine hydroxylase locus and evaluation of its potential application in prenatal diagnosis of phenylketonuria in Chinese. AB - Using a human phenylalanine hydroxylase cDNA probe, the restriction fragment length polymorphisms at the human phenylalanine hydroxylase locus have been determined with the restriction enzymes BglII, PvuII, EcoRI + BamHI, MspI, XmnI, HindIII and EcoRV. The frequency of the observed heterozygosity of the restriction site polymorphisms at this locus in a Chinese population is approximately 54%, which is significantly lower than that in Caucasians. No DNA rearrangement or deletion of the phenylalanine hydroxylase locus was detected among mutant phenylalanine hydroxylase genes in seven Chinese classical phenylketonuria (PKU) families. Haplotype analysis of these seven families revealed that the mutant alleles belonged to five different haplotypes, i.e. haplotype 4, 11 and three unreported haplotypes. The majority of normal and mutant phenylalanine hydroxylase genes are confined to hyplotype 4. These results indicate that approximately 42% of Chinese PKU families are informative for prenatal diagnosis of PKU when eight restriction sites linked to the phenylalanine hydroxylase locus are examined. PMID- 2563989 TI - Sulfasalazine therapy in inflammatory bowel disease. AB - Sulfasalazine currently is the most commonly prescribed medication for inflammatory bowel disease. Insights into the pharmacokinetics, their side effects and mechanism and their possible mode of action has led to rational basis for its clinical use and the development of new related agents. This review summarizes these aspects and discusses the future role of sulfasalazine in the face of new oral preparations. PMID- 2563990 TI - Topical salicylate therapy (4-ASA and 5-ASA enemas). AB - Topical therapy with amino salicylate enemas is the treatment of choice for proctitis, proctosigmoiditis, and left-sided ulcerative colitis. Adjunctive sulfasalazine therapy is often required. Relapse rates of 80 per cent can be expected within 3 months of discontinuing therapy. PMID- 2563992 TI - Spinal anesthesia for obstetrics. PMID- 2563991 TI - Antibiotics and inflammatory bowel diseases. AB - In evaluating the medical literature dealing with antibiotics and inflammatory bowel diseases, I cannot help but recall the adage: "Those who have enthusiasm have no controls and those who have controls have no enthusiasm." There just are not enough data to justify the use of most antibiotics in the treatment of most patients with Crohn's disease or ulcerative colitis. An increasing body of data does support the use of metronidazole in selected patients with Crohn's disease, especially those with perianal disease or fistulae. However, this drug has important side effects that may preclude its long-term use. Other antibiotics have been inadequately tested and there is not adequate evidence to support their use. PMID- 2563993 TI - Takayasu's arteritis. AB - Takayasu's arteritis, an inflammatory and obliterative disease of medium and large arteries, is classified as a giant cell arteritis. It has a predilection for the aortic arch and its branches and the pulmonary arteries. Unlike atherosclerotic vascular diseases, Takayasu's arteritis affects young women primarily. Early symptoms may be mainly systemic and may resemble polymyalgia rheumatica. The etiology is unknown, but infectious agents and autoimmunity have been suggested to play a role. Diagnosis is based on symptoms, physical findings, and angiographic findings, because tissue diagnosis rarely is feasible. The two cases reported here represent contrasting phases of Takayasu's arteritis. PMID- 2563994 TI - Maintaining mosquito cell lines at high temperatures: effects on the replication of flaviviruses. AB - The upper thermal limit for maintenance of eleven mosquito cell lines was studied. Although most cell lines could be grown at 32 degrees C to 34 degrees C, Anopheles stephensi cell line could be maintained at 37 degrees C. At higher temperatures initial growth rate was higher, but yield of cells after about a week of incubation was lower than at the standard temperature (28 degrees C). Replication of several flaviviruses in Aedes albopictus cell cultures adapted to 34.5 degrees C was faster, and viral titers were higher than at 28 degrees C. PMID- 2563995 TI - Rat kidney proximal tubule cells in defined medium: the roles of cholera toxin, extracellular calcium and serum in cell growth and expression of gamma glutamyltransferase. AB - A culture system is described in which rat kidney proximal tubule epithelial cells (RPTE) can be prepared with good yield and high viability and grown in culture under serum-free conditions. The cells require EGF, insulin, cholera toxin and either 1% dialyzed serum or a complex of bovine serum albumin with oleic acid (BSA/OA). The cells can be maintained for long periods of time and express several markers for RPTE. The cells have both alkaline phosphatase and gamma-glutamyltransferase activity and respond to parathyroid hormone but not vasopressin. The specific activity of gamma-glutamyltransferase decreases when the cells begin to grow, but increases when they reach confluence. Extracellular calcium plays a role in the induction of gamma-glutamyltransferase in confluent cells. Cells grown in media containing low calcium, i.e. less than 0.4 mM, have reduced specific activity of gamma-glutamyltransferase. Extracellular calcium also alters the morphology of the cells in that cells grown in low calcium are single cells or loose clusters suggesting poor cell-cell contact. When the calcium is raised to 1.0 mM, the cells change their shape and organization to adopt the morphology of cells maintained continuously in 1.0 mM calcium. The cells can be passaged onto plastic surfaces which have been coated with collagen but cannot be subcultured on uncoated or serum coated plastic. This culture system will be a useful model for the investigation of renal carcinogenesis and the role of cell proliferation in that process. PMID- 2563997 TI - The groES and groEL heat shock gene products of Escherichia coli are essential for bacterial growth at all temperatures. AB - The products of the groES and groEL genes of Escherichia coli, constituting the groE operon, are known to be required for growth at high temperature (42 degrees C) and are members of the heat shock regulon. Using a genetic approach, we examined the requirement for these gene products for bacterial growth at low temperature (17 to 30 degrees C). To do this, we constructed various groES groEL heterodiploid derivative strains. By inactivating one of the groE operons by a polar insertion, it was shown by bacteriophage P1 transduction that at least one of the groE genes was essential for growth at low temperature. Further P1 transduction experiments with strains that were heterodiploid for only one of the groE genes demonstrated that both groE gene products were required for growth at low temperature, which suggested a fundamental role for the groE proteins in E. coli growth and physiology. PMID- 2563996 TI - Identification and characterization of the genes encoding the type 3 and type 1 fimbrial adhesins of Klebsiella pneumoniae. AB - Strains of Klebsiella pneumoniae are known to express two morphologically and functionally distinct filaments, the type 3 and the type 1 fimbriae. The gene (mrkD) encoding the adhesion of K. pneumoniae type 3 fimbriae was identified by transcomplementation analysis with the pap fimbrial gene cluster of Escherichia coli. The nucleotide sequence of the mrkD gene was determined. In addition, the determinant coding for the K. pneumoniae type 1 fimbrial adhesion was identified, and its nucleotide sequence was deduced. The predicted amino acid sequences of the K. pneumoniae adhesion proteins are compared, and similarities with the major fimbrial structural proteins (MrkA and FimA) are discussed. PMID- 2563998 TI - Rhizobium meliloti 1021 has three differentially regulated loci involved in glutamine biosynthesis, none of which is essential for symbiotic nitrogen fixation. AB - We have cloned and characterized three distinct Rhizobium meliloti loci involved in glutamine biosynthesis (glnA, glnII, and glnT). The glnA locus shares DNA homology with the glnA gene of Klebsiella pneumoniae, encodes a 55,000-dalton monomer subunit of the heat-stable glutamine synthetase (GS) protein (GSI), and complemented an Escherichia coli glnA mutation. The glnII locus shares DNA homology with the glnII gene of Bradyrhizobium japonicum and encodes a 36,000 dalton monomer subunit of the heat-labile GS protein (GSII). The glnT locus shares no DNA homology with either the glnA or glnII gene and complemented a glnA E. coli strain. The glnT locus codes for an operon encoding polypeptides of 57,000, 48,000, 35,000, 29,000, and 28,000 daltons. glnA and glnII insertion mutants were glutamine prototrophs, lacked the respective GS form (GSI or GSII), grew normally on different nitrogen sources (Asm+), and induced normal, nitrogen fixing nodules on Medicago sativa plants (Nod+ Fix+). A glnA glnII double mutant was a glutamine auxotroph (Gln-), lacked both GSI and GSII forms, but nevertheless induced normal Fix+ nodules. glnT insertion mutants were prototrophs, contained both GSI and GSII forms, grew normally on different N sources, and induced normal Fix+ nodules. glnII and glnT, but not glnA, expression in R. meliloti was regulated by the nitrogen-regulatory genes ntrA and ntrC and was repressed by rich N sources such as ammonium and glutamine. PMID- 2563999 TI - Induced multidrug resistance in murine leukemia L1210 and associated changes in a surface-membrane glycoprotein. AB - The aim of this study was to find out whether only resistant cells of the "multidrug-resistant" phenotype show the described changes of plasma membrane glycoprotein (170 kDa) or whether resistant cells that do not express this phenotype reveal corresponding results. Doxorubicin-resistant (L1210dox) and daunorubicin-resistant L1210 ascites tumor cells (L1210dnr) (multidrug-resistant tumor cells) were therefore compared with cytosine-arabino-side-resistant (L1210AraC) and cyclophosphamide-resistant L1210 ascites tumor cells (L1210ctx) (not multi-drug-resistant tumor cells). The resistant cell lines were generated in vivo in tumor-bearing mice and the resistance to cytostatic agents was evaluated in vivo and in vitro. Using the accumulation assay with rhodamine-123, the multidrug resistance can be detected. In order to determine alterations in the plasma membranes we used the monoclonal antibodies 265/F4 and C219, which were prepared against the membrane glycoprotein P170 (170 kDa) in colchicin resistant Chinese hamster ovary cells. The results demonstrate that L1210dox and L1210dnr tumor cells show an intense immunostaining by the streptavidin/biotinylated-peroxidase-complex method and by the streptavidin/biotin/phycoerythrin immunofluorescence method. In contrast no specific immunostaining was observed in parental (sensitive) and L1210AraC or L1210ctx tumor lines. The results were confirmed by immunoblotting. To determine whether multidrug-resistant DNA sequences were expressed in the multidrug resistant tumor cells, Northern blots with RNA od sensitive and resistant cells were performed using the clone pcDR1.5. Elevated RNA levels were detected only in resistant cells with the multidrug-resistant phenotype. Thus, the results of this study demonstrate that only resistant cells with the multidrug-resistant phenotype show an increased expression of the membrane 170-kDa glycoprotein. PMID- 2564000 TI - H2-antagonists and carmustine. AB - The highly metabolized nitrosourea carmustine (BCNU) is an anticancer agent which alkylates DNA and is metabolized to both active and inactive species by cytochrome P-450 enzymes. Other highly metabolized anticancer drugs have altered toxicities when some histamine H2 antagonists are coadministered. To test this hypothesis with BCNU, DBA/2J male mice were given a single injection of cimetidine (CMT 100 mg/kg) or ranitidine (RNT 25 mg) at various times up to 30 min before, or up to 60 min after a BCNU injection. Spleen colony assays for normal bone marrow stem cell viability showed enhanced toxicity for BCNU when CMT was administered concomitantly. In P-388 leukemia-bearing DBA/2J mice, both CMT and RNT significantly enhanced the antitumor effects of BCNU doses of 30 mg/kg. Pharmacokinetic analyses of BCNU elimination in Cd-1 mice showed marked prolongation of BCNU elimination and increased (BCNU concentration) x time products when CMT was concomitantly administered. These results demonstrate that enhanced BCNU bone marrow toxicity and antitumor activity is produced by CMT. The effect appears to be related to impaired drug clearance when the two agents are administered concurrently. RNT slightly enhanced BCNU antileukemic effects, but it did not significantly alter BCNU myelotoxicity nor drug elimination patterns. PMID- 2564001 TI - Correlation between lectin binding and clinical factors in seminoma patients. AB - The binding of a panel of lectins to histological sections of seminomas was studied. The findings were correlated with different clinical parameters. Concanavalin A and wheat germ agglutinin stained all seminomas whereas horse gram (DBA) and peanut agglutinin did not stain the tumor cells. A varying staining pattern was found with lectins from castor bean (RCA I), soy bean (SBA) and gorse (UEA I) indicating a heterogeneity of the tumor cell population. In the seminomas that were derived from undescended testis there were more cases that showed positive staining with soy bean agglutinin, which shows that the intra-abdominal location of the seminoma might cause changes in the cellular metabolism resulting in glycoconjugates different from those in descended tumors. No correlation was found between the lectin staining and the prognosis, stage or metastasis of the tumor. PMID- 2564002 TI - Cloning of cDNAs encoding two related 100-kD coated vesicle proteins (alpha adaptins). AB - Coat proteins of approximately 100-kD (adaptins) are components of the adaptor complexes which link clathrin to receptors in coated vesicles. The alpha adaptins, which are found exclusively in endocytic coated vesicles, separate into two bands on SDS gels, designated A and C (Robinson, M. S., 1987. J. Cell Biol. 104:887-895). Two distinct cDNAs (sequences 1 and 2) encoding the two alpha adaptins were cloned from a mouse brain cDNA library. Southern blotting indicates that there is one copy of each of the two alpha-adaptin genes, and that there are no additional closely related genes. Based on the size of the predicted protein products of the two genes (108 and 104 kD), the relative abundance of the two messages in brain and liver, and the reactivity of a sequence 1 fusion protein with different antibodies, it was possible to conclude that sequence 1 codes for A and sequence 2 for C. The two protein sequences are strikingly homologous to each other (84% identical amino acids), the major difference being an additional stretch of 41 amino acids, rich in prolines and acidic residues, inserted into the COOH-terminal half of A. In situ hybridization carried out on mouse brain sections indicates that the same cell type may express both transcripts, but that their relative expressions vary. Antipeptide antibodies are now being raised to find out whether the proteins are localized in functionally distinct populations of endocytic coated vesicles. PMID- 2564004 TI - Postischemic administration of idazoxan, an alpha-2 adrenergic receptor antagonist, decreases neuronal damage in the rat brain. AB - The effect of an alpha-2 receptor antagonist, idazoxan, on ischemic neuronal damage in the hippocampus and neocortex was studied in rats following 10 min of forebrain ischemia. Idazoxan was given 0.1 mg/kg i.v. immediately after recirculation, followed by 48 h of continuous infusion at a rate of 10 micrograms/kg/min. A histopathological examination of the CA1 region of the dorsal hippocampus and neocortex from each hemisphere was made on paraffin embedded sections following 7 days of survival. In ischemic animals receiving an infusion of saline, 71% of the neurons in the hippocampal CA1 region were degenerated. In contrast, in the idazoxan-treated animals only 31% of the neurons were irreversibly damaged (p less than 0.01). We conclude that postischemic administration of the alpha-2 antagonist idazoxan protects neurons against damage following cerebral ischemia. Rapid postischemic administration of alpha-2 adrenergic receptor antagonists could be an effective treatment after stroke and cardiac arrest. PMID- 2564003 TI - Formation of coated vesicles from coated pits in broken A431 cells. AB - Biochemical and morphological techniques were used to demonstrate the early steps in the endocytosis of transferrin in broken A431 cells. After binding 125I transferrin, the cells were broken by scraping and then warmed. 125I-transferrin became inaccessible to exogenous anti-transferrin antibody providing a measure of the internalization process. Parallel morphological experiments using transferrin coupled to horseradish peroxidase confirmed internalization in broken cells. The process was characterized and compared with endocytosis in intact cells and showed many similar features. The system was used to show that both the appearance of new coated pits and the scission of coated pits to form coated vesicles were dependent on the addition of cytosol and ATP whereas invagination of pits was dependent on neither. PMID- 2564005 TI - 1 Alpha,25-dihydroxyvitamin D3 receptor distribution and effects in subpopulations of normal human T lymphocytes. AB - Receptors for 1 alpha,25-dihydroxyvitamin D3 [1,25-(OH)2D3] are expressed upon activation of human lymphocytes, and the hormone inhibits in vitro the proliferation of mitogen-activated lymphocytes. In this study we examined the distribution of the 1,25-(OH)2D3 receptor protein in the two major subsets of T lymphocytes (T helper and T suppressor cells) and the effect of the hormone on their respective rates of proliferation. We activated normal lymphocytes in the presence of monocytes with phytohemagglutinin and subsequently isolated the T helper (T4-positive) and the T suppressor (T8-positive) subsets using monoclonal antibodies and complement-mediated lysis. In parallel experiments, we first isolated monocyte-depleted T4 and T8 cells and then activated them using phytohemagglutinin and a phorbol ester. Using either approach we found that both T4 and T8 lymphocytes expressed the 1,25-(OH)2D3 receptor protein upon activation. The concentration of this protein, its affinity for the ligand (Kd, approximately 10(-10) mol/L), and its sedimentation characteristics (S = 3.3) were indistinguishable in the two T cell subsets. Furthermore, the time kinetics of expression of the receptor after activation were very similar in the two subsets. Nevertheless, 1,25-(OH)2D3 inhibited in a dose-dependent fashion the rate of proliferation of the helper subset, but had no effect on the proliferation of suppressor cells. The finding of a dissimilar effect of 1,25 (OH)2D3 on the proliferation of the T helper and T suppressor cells despite their indistinguishable receptor status suggests that the 1,25-(OH)2D3 receptors of the T cells might not be involved in the effects of the hormone on T-cell proliferation, and that the 1,25-(OH)2D3-induced inhibition of mitogen-activated T4 cell proliferation could be mediated indirectly. PMID- 2564007 TI - Histometric studies on cellular infiltrates of tuberculin tests in patients with haemophilia. AB - The number of microanatomical location of CD4 and CD8 lymphocytes, of cells bearing receptors for I12 and transferrin, and of monocyte/macrophages in the dermis at the site of a tuberculin test were measured in 13 patients with haemophilia (10 seronegative and three seropositive for human immunodeficiency virus (HIV]. The overall density of lymphocytes in the perivascular and diffuse parts of the infiltrate was similar to that reported in other groups of subjects without evidence of immunosuppression. The CD4:CD8 ratio of the infiltrating lymphocytes throughout the section showed an inverse relation with clotting factor consumption. There was no significant change in the CD4:CD8 ratio in the diffuse infiltrate at various levels into the dermis in tuberculin reactions in patients with haemophilia, unlike healthy controls and other groups with no evidence of immunosuppression, who have previously been shown to have increasing CD4:CD8 ratio with increasing depth into the dermis. The number of cells bearing receptors for I12 and transferrin and of monocyte/macrophages was related to total lymphocyte density in the infiltrate. There was no evidence of serious impairment of the cell mediated response to a long term recall antigen, but the relatively low preponderance of CD4 lymphocytes in the diffuse infiltrate, particularly in the deeper dermis, may be the earliest indicator of impending immunosuppression. PMID- 2564008 TI - A revised interpretation of the TRH test results in female depressed patients. Part II: Prolactin responses. Relationships with sex hormones, corticosteroid state, age, monoamines and amino acid levels. AB - Prolactin (PRL) levels were recorded in baseline conditions and 20 and 60 min after thyrotropin releasing hormone (TRH) administration (200 micrograms i.v.) in 60 depressed females categorized according to DSM-III. Peak PRL responses were significantly (r = 0.727, P less than 0.001) correlated with their baseline levels. Consequently, the PRL responses to TRH were largely predicted by baseline PRL levels. It was suggested that the PRL responses to TRH consisted of two parts. The first component was a relative exaggeration of basal PRL, reflecting the basal activity of the hormone. The second component was the residual response. This part was estimated by partialling out the relative effects of basal PRL on peak PRL responses by means of regression analysis. Basal PRL and residual PRL responses were uninformative for major depression. Post-menopausal females showed significantly reduced basal PRL levels. There was a significant negative correlation between basal PRL and follicle stimulating hormone levels, age and post-dexamethasone cortisol values. The residual PRL responses were negatively correlated with free triiodothyronine levels and positively with serotonergic variables, i.e., 5-hydroxyindoleacetic acid in 24-h urine and the ratio L-tryptophan/competing amino acids. PMID- 2564006 TI - Class specific antibody response to gonococcal infection. AB - An enzyme immunoassay was used to determine IgM, IgG, and IgA antibodies to gonococcal pili in 68 patients with uncomplicated gonorrhoea, 35 women with pelvic inflammatory disease, and in 115 normal controls. A clear difference in response rate in all three antibody classes between patients with gonorrhoea and healthy controls was evident. Among women with gonorrhoea, the magnitude of antibody response was higher than among men with gonorrhoea, especially in the IgM class. No major differences were found in the overall distribution of serological findings between women with uncomplicated gonorrhoea and those with gonococcal pelvic inflammatory disease. Among this last group, however, high IgM antibody levels in acute phase sera were significantly associated with the isolation of Neisseria gonorrhoeae in the upper genital tract. PMID- 2564009 TI - High hepatic gamma-glutamyltransferase (gamma-GT) activity with normal serum gamma-GT in children with progressive idiopathic cholestasis. AB - gamma-Glutamyl transferase (gamma-GT) was assayed in the serum and liver biopsies of children affected with either progressive idiopathic cholestasis (PIC, Byler's disease), or other types of cholestatic (biliary atresia, cholestasis of various origins) and non-cholestatic diseases. The mean liver gamma-GT activity was increased significantly only in PIC and biliary atresia. In contrast, the serum gamma-GT activity, raised in children with evident damage to the main bile ducts or to the interlobular bile ducts, was normal in children with PIC. Although the mechanism for such a discrepancy between high liver and normal serum gamma-GT activities in PIC is still speculative, this peculiarity could prove to be of use in leading to a better understanding of the disease. PMID- 2564011 TI - Effect of ursodeoxycholic acid treatment on alanine aminotransferase and gamma glutamyltranspeptidase serum levels in patients with hypertransaminasemia. Results from a double-blind controlled trial. AB - The ability of ursodeoxycholic acid (UDCA, 600 mg/day) to lower alanine aminotransferase (ALT) blood levels in blood donors rejected for donation because of fluctuating hypertransaminasemia was evaluated in a randomized, controlled, double-blind clinical trial vs. placebo. All subjects with ALT values at least twice the normal upper limit in at least two out of three previous checks (the last one not more than 1 month previously) were admitted to the study. Checks were carried out 1, 2 and 3 months after the admission. 59 out of 65 patients completed the study. Although all patients were asked to abstain from alcohol, more than 50% of them in both groups had basal gamma-glutamyltransferase (gamma GT) values higher than normal. After 1 month of treatment and throughout its duration, UDCA was effective in lowering ALT in all patients (30% decrease with respect to the basal value) and, especially, in lowering gamma-GT in those patients with elevated levels (50% decrease with respect to the basal value). This decrease was significantly different from the spontaneous 10% decrease of the ALT and gamma-GT levels observed in the placebo group. 3 months after suspension of therapy a rebound of both ALT and gamma-GT to values comparable to the basal ones or even higher was found only in UDCA-treated patients. We conclude that the short-term administration of UDCA is free of hepatotoxic effects and could be useful in lowering ALT and gamma-GT serum levels. The real significance of UDCA treatment in the natural history of chronic liver diseases deserves further investigation. PMID- 2564012 TI - Computer-aided surveillance of surgical infections and identification of risk factors. AB - A continuous record of postoperative surgical infections was carried out by electronic data processing (EDP) of 4340 orthopaedic and general operations. The overall infection rate was 6.3%, ranging from 2.3% (clean wounds) to 27.1% (dirty wounds). The corresponding deep infection rates were 1.6%, 0.4% and 4.6%. Employing a multiple logistic regression analysis, 10 risk factors were evaluated. Factors found to be significant were: wound contamination, department, duration of operation, date of operation and age, and in addition for the department of general surgery: surgeon, planning of operation, length of preoperative stay and anatomic groups. A statistical model for identification of risk patients is described. Postoperative stay was on average 20.5 days longer in infected patients. We find that EDP-recording may result in an annual cost reduction of at least 175,000 pounds for our hospital. PMID- 2564013 TI - Studies on the prevalence of Pseudomonas aeruginosa in a Jordanian hospital. PMID- 2564010 TI - Effect of cimetidine, ranitidine, famotidine and omeprazole on hepatocyte proliferation in vitro. AB - Recently reports have indicated that both cimetidine and ranitidine delay cell proliferation in rats following 70% partial hepatectomy and result in an increased mortality following this procedure. The present study was designed to determine whether three H2 blocking agents (cimetidine, ranitidine, famotidine) and a new, powerful antisecretory drug (omeprazole) specifically influence hepatocyte proliferation in primary culture. Hepatocytes were isolated from livers of normal male rats by the standard collagenase perfusion technique. Hepatic DNA synthesis and percent of labelled nuclei were determined after 48 h incubation. Hepatocytes in culture were incubated with the H2 blocking agents and omeprazole or with different concentrations of serum obtained from sham-operated or 70% hepatectomized rats treated or not with the same agents. Rats were injected intraperitoneally at 8:00 a.m. on two consecutive days. In hepatectomized rats, the first dose was injected at 8:00 a.m. immediately after surgery, the second, 24 h later. The serum of sham-operated or 70% hepatectomized rats that did not receive drugs served as control. No changes in DNA synthesis, percentage of labelled nuclei and transaminase were detected when the agents were added to the hepatocytes in culture at concentrations within the effective pharmacological dosage and 30 times higher. Similarly, no changes in these parameters were obtained when different concentrations of serum obtained from sham-operated rats treated with H2 blocking agents or omeprazole were added to the basal culture medium. However, a significant inhibition of DNA synthesis and of percentage of labelled nuclei was observed when hepatocytes were incubated in the presence of serum from 70% hepatectomized rats that had been treated with cimetidine or with ranitidine. The serum of 70% hepatectomized rats treated with famotidine and omeprazole had no effect on hepatocyte proliferation in vitro. No effect on transaminase was found in these conditions. PMID- 2564014 TI - Invasive aspergillus infection: possible non-ward common source within the hospital environment. AB - Six immunocompromised patients housed in widely separated portions of a hospital campus developed invasive aspergillosis during a single month. This represented a significant increase (P less than .001) in the number of cases observed over the 3 years which included this event (19 cases in 36 months). Epidemiological investigation suggested that this cluster of cases was due to a common source outbreak related to construction activity in a central radiology suite serving the hospital. Such non-ward exposures to nosocomial hazards are becoming increasingly important for immunosuppressed hosts. PMID- 2564015 TI - Infection risks from electrically operated breast pumps. AB - Using formula milk seeded with Staphylococcus epidermidis as indicator, bacteria were observed to pass beyond the collecting bottles of three electrically operated breast pumps. Bacteria were recovered from sites distal to the level of visible contamination, the incidence increasing with repeated use of the apparatus. Despite use of a sterile collecting bottle, retrograde contamination of freshly collected milk may occur from previously contaminated components of the pumps. The results suggest that a terminal in-line air filter is essential to ensure aerosols containing potentially pathogenic bacteria do not contaminate the suction source or be emitted to the environment with the exhaust air. Where a single pump is used by more than one person adequate sterilization of all removable components is essential. PMID- 2564016 TI - Methods for calculating the efficiency of bacterial surface sampling techniques. AB - A mathematical model is described which may be used to assess the efficiency and consistency of a surface sampling method as well as estimate the concentration of bacteria on the site prior to sampling. PMID- 2564017 TI - The use of intravenous cannulae and the occurrence of thrombophlebitis. AB - The occurrence of thrombophlebitis in a coronary care unit was studied in relation to the use of short plastic intravenous cannulae. The incidence of thrombophlebitis was 51% in cases where cannulae were used for continuous infusion of glucose 5% and 13% for cannulae which were locked after the injection of heparin. Only one case of infectious thrombophlebitis was seen. The other cases of thrombophlebitis had a chemical or mechanical aetiology. Replacement of glucose 5% by a NaCl 0.9% solution for continuous infusion reduced the incidence of thrombophlebitis to 33%. Heparin-locked cannulae, to provide rapid access to the patient's circulation, proved to be a safe alternative to continuous infusion. PMID- 2564018 TI - Microbial contamination of hospital showers and shower water: the effect of an automatic drain valve. AB - The installation of a valve to drain away water on the completion of showering was found to have no significant effect on the numbers of microorganisms in shower water and on the internal surfaces of shower fittings. PMID- 2564019 TI - Measurement of bacterial and fungal air counts in two bone marrow transplant units. AB - We evaluated air contamination with bacteria and fungi in a transplantation unit, successively housed in two buildings. Bacterial air contamination was least in laminar air flow rooms, and reduced in ultraclean air rooms in comparison with conventional rooms. Similar results were obtained with culture of air for fungi. PMID- 2564020 TI - Evaluation of ethylene oxide sterilization of tissue implants. AB - The ability of ethylene oxide (ETO) gas to penetrate tissue matrices was evaluated using Bacillus subtilis var niger (NCTC 10073) spore strips sandwiched between sheets of various tissues exposed to ETO for varying periods. The decimal reduction times (min) obtained using commercially prepared spores were: 7.97 for naked spores, 7.34 between amnion, and 6.39 between amnio-chorion, indicating complete penetration of the matrix by the gas. Similar conclusions were reached using skin and dura mater. The possible protective effect of serum was evaluated by drying spores suspended in serum in different ways, followed by exposure to the gas. Decimal reduction times obtained were: 8.67 (dried at 37 degrees C); 8.09 (dried at 55 degrees C) and 8.97 (freeze-dried) confirming slight protection. However, in all cases sterilization was achieved following 100 min ETO exposure, giving a 2.5 times safety factor for the commercial ethylene oxide sterilizer. PMID- 2564021 TI - Lowered prevalence of infection with lactulose therapy in patients in long-term hospital care. AB - Over a 6-month, retrospective study period, regular lactulose therapy resulted in a significant reduction in the prevalence of UTIs when compared to a control group receiving no lactulose (P less than 0.005). The overall number of patients contracting infections was also reduced (P less than 0.25). There was a parallel reduction in the prescription of antibiotics (P less than 0.05) and in the number of patients requiring antibiotic therapy (P less than 0.005). There was no alteration in faecal continence with lactulose therapy and no urinary catheters were used in either group. In two additional groups of patients, low dose lactulose therapy for 6 months and a shorter duration of lactulose therapy for under 2 months, had no significant effect on the prevalence of UTIs. This study was retrospective and the results were both unexpected and do not explain the mode of action of lactulose. A controlled prospective study involving larger patient numbers is required before any realistic therapeutic recommendations can be made. PMID- 2564022 TI - Routine cleaning and the elimination of Campylobacter pylori from endoscopic biopsy forceps. AB - The effect of routine cleaning in removing Campylobacter pylori from the biopsy forceps of endoscopes has been examined in a series of 50 patients. Campylobacter pylori was isolated from the biopsies of 15 of the patients, while one of the 50 biopsy forceps washings yielded the organism after routine cleaning. This study suggests that there is a small chance of transmitting C. pylori by endoscopic equipment if cleaning is the only method of decontamination adopted. PMID- 2564023 TI - Laundering of nurses' dresses at home. PMID- 2564024 TI - Appropriate disposal of wound dressings. PMID- 2564025 TI - MIC tests are not suitable for assessing antiseptic handwashes. PMID- 2564026 TI - Effect of chlorhexidine on bacterial pathogenicity. PMID- 2564027 TI - MRSA and environmental cleanliness. PMID- 2564028 TI - Prevalence of antibiotic use in a hospital in Brazil. PMID- 2564029 TI - HIV-induced immunodeficiency. Relatively preserved phytohemagglutinin as opposed to decreased pokeweed mitogen responses may be due to possibly preserved responses via CD2/phytohemagglutinin pathway. AB - We studied the proliferative response of PBL to the mitogens PHA and PWM and Candida albicans Ag in 301 HIV seropositive homosexual men, of whom 55 had AIDS. The responses to PHA were reduced only in the clinically ill HIV seropositive subjects. In contrast, the responses to PWM were profoundly reduced in most HIV seropositive subjects including the asymptomatic group. Further analysis of 16 HIV seropositive subjects showed that the proliferative responses were reduced in both CD4 and CD8 T cell subsets. A total of 15 HIV seropositive individuals with low responses to PWM, of whom seven had AIDS and eight controls were chosen for the following studies. Expression of T3, Ti, delta receptors, and CD2 was investigated and showed an increased percentage of CD2 receptors positive cells in HIV seropositive subjects without AIDS. The proliferative responses of PBL to stimulation with PHA, PWM, antibodies to CD3, or antibodies to CD2 were investigated and showed significant correlation in controls, whereas in contrast, only the responses to PHA and CD2ab correlated in patients with AIDS. The proliferative responses to CD2ab and CD3ab in controls were larger than the responses to both PHA and PWM. In patients, these responses were less suppressed than the responses to PWM indicating that stimulation with mitogens is more complex than a simple stimulation of Ti/T3 and CD2 receptors. Further investigations were done on resting T cells, i.e., lymphocytes depleted of macrophages and pre-activated cells. Addition of PHA to these cells resulted in preactivation with expression of IL-2R (CD25) but not in proliferation. In contrast, addition of PHA plus SRBC, which bind to the CD2 receptors caused IL-2R expression, IL-2 production, and proliferation. Addition of PWM + SRBC did not result in proliferation. A comparison of the responses to PHA + SRBC of resting T cells from 26 HIV seropositive individuals, of whom seven had AIDS and 12 seronegative controls, showed that these responses were normal or only slightly decreased in the 19 seropositive men without AIDS whereas it was decreased in AIDS patients. Nevertheless, all AIDS patients showed clear-cut responses in this assay. Thus, the discrepancy between responses to PHA and PWM may be explained by an at least partially preserved function of the PHA/CD2-dependent pathway. We suggest that the defect induced by the HIV infection primarily concerns T3/Ti induced responses. PMID- 2564030 TI - An improved method for the treatment of data from DNA strand break measurements using filter elution techniques with an internal standard. PMID- 2564032 TI - Free radical scavenging activity of papaya juice. AB - Papaya juice is an efficient scavenger of highly reactive hydroxyl radicals (OH.) formed during 60Co irradiation of water. The OH. radicals were detected by the electron spin resonance (ESR) technique of spin trapping using DMPO (5,5-dimethyl 1-pyrroline-N-oxide) or by a colorimetric assay in which salicylate is converted into polyhydroxybenzoic acids. Papaya juice is also able to quench the ESR signal of a stable free radical (TEMPOL) and the ESR signal of the DMPO-OH adduct. The active substance(s) in papaya juice are heat-stable, dialyzable, and soluble in water but not in lipid solvents. The active agents do not appear to be ascorbate, tocopherol, or carotenoids. PMID- 2564031 TI - Matrix-isolation of H-induced free radicals from purines in acidic glasses. AB - H.(D.)-atoms produced by photo-oxidation (lambda = 254 nm) of Fe2+ in acidic glasses (6 mol dm-3 H2SO4/H2O or D2SO4/D2O) at 77K were allowed to react with the purine bases adenine, guanine, hypoxanthine and xanthine as well as with ribo- and deoxyribosides (-tides) of adenine and guanine by annealing to 110-130 K. The ensuing free radicals were studied by electron spin resonance (ESR) spectroscopy at 77 K. Individual radical species were assigned by simulation of patterns isolated from thermal annealing studies up to 180 K. It is shown that H.-atom reaction with the bases produces C2- and C8-addition radicals for adenine and C8 addition species for guanine. Guanine is also photo-oxidized directly in the glass, producing a cation or its deprotonated successor species. In the nucleosides (-tides) of both bases, H. atoms were found to abstract hydrogen from carbon sites C1', C2' or C3', and C5' for ribosides and C1', C2', and C5' for deoxyribosides (-tides), respectively. PMID- 2564033 TI - Radiation-induced crosslinking between poly(deoxyadenylic-deoxythymidylic acid) and tripeptides containing aromatic residues. AB - OH.-induced covalent peptide-nucleotide adducts have been isolated by reverse phase chromatography from the enzymic hydrolyzates of gamma-ray irradiated solutions containing double-stranded poly(deoxyadenylic-deoxythymidylic acid) and one of the tripeptides, lysyl-tryptophyl-lysine or lysyl-tyrosyl-lysine. Numerous compounds were formed, resulting presumably from different modes of radical addition. All isomers appeared to have the same general structure peptide d(ApTpA), based mostly on double-labelling experiments of bases and phosphate groups in DNA. The major adduct fraction obtained from Lys-Trp-Lys and poly(dA dT) was purified to homogeneity by sequential reverse-phase and ion-exchange chromatography, and characterized spectrally. The pattern of acid and alkaline hydrolysis suggests that thymine is the site of peptide-nucleotide binding in this particular adduct fraction. PMID- 2564034 TI - Similar lethal effect in mammalian cells for two radioisotopes of copper with different decay schemes, 64Cu and 67Cu. AB - The decays of 64Cu incorporated in human malignant (A549) or monkey nonmalignant (CVI) cells lead to cell death. When plotted as a function of the radioactivity introduced in the growth medium (microCi/ml at t = 0), the residual colony forming capability decreases exponentially. The slope of the corresponding curve is steeper for A549 than for CV1 cells. Different data show that the cellular lethal event is a consequence of 64Cu transmutation and not of the irradiation by the simultaneously emitted beta- and beta+ particles. Liquid holding results show that the lethal event is irreparable. The decays of 67Cu, another radioisotope of copper, lead to cell death with the same exponential survival curve and the same lethal efficiency as for 64Cu, in spite of their different decay schemes. The lethal efficiency of both copper isotopes is close to that of 125I utilized in the form of iododeoxyuridine under the same experimental conditions as 64Cu and 67Cu. The high lethal efficiency of radioactive copper transmutations raises questions about the role in DNA functioning of copper atoms known to be present in trace amounts in this macromolecule. The lethal consequence of radioactive copper transmutations suggests that the copper atoms bound to DNA are essential for cellular functioning. PMID- 2564035 TI - Radiation-induced DNA damage and repair in quiescent and proliferating human tumor cells in vitro. AB - The purpose of this study was to examine radiation-induced DNA strand breakage and repair in quiescent and proliferating human tumor cells in vitro and determine their relationship to radiation sensitivity and potentially lethal damage repair (PLDR). Using centrifugal elutriation we have isolated from fed plateau-phase cultures of HEp-3 human squamous carcinoma cells, relatively pure populations of quiescent and proliferating cells. This was confirmed by both [3H] thymidine labelling and acridine orange (AO) staining with flow cytometry. Quiescent cells were more sensitive to ionizing radiation (Do = 0.97 Gy) than were proliferating cells (Do = 1.28 Gy). However, quiescent cells showed higher repair of potentially lethal damage (PLDR) than did proliferating cells. Repair of single-strand breaks (ssb) and double-strand breaks (dsb) as measured by filter elution did not differ significantly between quiescent and proliferating cells. For both populations, ssb and dsb repair kinetics and final damage remaining were the same, suggesting that repair of DNA strand breaks is not entirely responsible for the difference in radiation sensitivity between quiescent and proliferating cells. PMID- 2564036 TI - Lethal effect and potentially lethal damage recovery in cultured mammalian cells irradiated by neutron-capture beams. AB - Cell killing and potentially lethal damage (PLD) recovery in Hela cells were examined after irradiation in suspension using a 10B neutron-capture beam. Thermal-neutron irradiation in the medium containing from 0.09 to 9 mM boron-10 (0.9-90 micrograms/g 10B, 90 per cent enriched boric acid) resulted in steeper survival curves than in the boron-free medium. The relative biological effectiveness (r.b.e.), expressed as the Do ratio, increased from 2.2 +/- 0.1 for the control without boron-10 to 5.4 +/- 0.3 in the medium containing 0.9 mM boron 10. The r.b.e. value was 5.2 +/- 0.5 at a boron-10 concentration of 9 mM, and did not increase when the boron-10 concentration was increased from 0.9 to 9 mM. It was suggested that the observed plateau of r.b.e. at boron-10 concentrations higher than 0.9 mM was due to the dominancy of the 10B(n, alpha)7Li reaction for dose accumulation. PLD recovery after the neutron irradiation containing 0.9 mM boron-10 was not observed, while a small effect was observed after irradiation in the thermal neutron beam, although much less than after gamma rays. The results show that the neutron-capture beam is an excellent beam for radiotherapy if the boron-10 concentration in tumour cells could be elevated to about 1 mM. PMID- 2564037 TI - Induction of tumour hypoxia post-irradiation: a method for increasing the sensitizing efficiency of misonidazole and RSU 1069 in vivo. AB - It is known that hydralazine can decrease blood flow to experimental murine tumours. A consequence of this, in the KHT sarcoma, is the induction of close to 100 per cent radiobiological hypoxia, which lasts for nearly 2 h following i.v. injection of 5 mg/kg hydralazine to the mouse. This phenomenon is exploitable in order to increase the apparent sensitizing efficiency of the nitroheterocyclic radiosensitizers, misonidazole and RSU 1069, and is demonstrated using the treatment schedule: sensitizer----60 min----X-rays----1 min----hydralazine. Such a strategy will first take advantage of the radiosensitizing properties of the nitroimidazole, then after irradiation the hydralazine should allow expression of the differential toxicity towards hypoxic cells known to occur with misonidazole and RSU 1069. Misonidazole gives an enhancement ratio (ER) of 1.3 at 100 mg/kg, rising to 2.0 at 1000 mg/kg. Where hydralazine is given after irradiation, no additional cell kill is observed with 1000 mg/kg. In contrast, at lower doses of misonidazole, hydralazine induces a substantial increase in cell killing such that the ER obtained with 100 mg/kg is the same as that achieved with 1000 mg/kg misonidazole when used alone with radiation. Similarly, 20 mg/kg RSU 1069 with radiation followed by hydralazine is equivalent to the radiosensitizing effect of 80 mg/kg RSU 1069 without hydralazine. In addition, doses of RSU 1069 that normally give no radiosensitization (5 or 10 mg/kg) produce substantial increases in cell killing when combined with hydralazine. PMID- 2564038 TI - Interaction of hyperthermia and radiation in tolerant and nontolerant HeLa S3 cells: role of DNA polymerase inactivation. AB - The activities of DNA polymerase alpha and beta were measured in tolerant and nontolerant HeLa S3 suspension cells. The heat-inactivation of the enzymes and their recovery when cells were incubated at 37 degrees C after the heat challenge was compared to the synergistic action of heat and radiation and its disappearance at the level of cell survival. Thermotolerant cells were radiosensitized by heat similarly to nontolerant cells, but the sensitization decreased more rapidly in the tolerant cells when time at 37 degrees C was allowed between the two treatments. For polymerase activities the extent of inactivation, as well as the kinetics of recovery, were similar in tolerant and nontolerant cells. The results show that the activities of DNA polymerase alpha and beta do not always correlate with the extent of heat radiosensitization. It is concluded that heat inactivation of these enzymes may not be taken as a general cause for the synergistic effect of hyperthermia and radiation. As an alternative mechanism, changes in nuclear protein binding due to cellular heating are suggested, since these correlate well with effects observed for radiosensitization under different experimental conditions, including the use of thermotolerant cells. PMID- 2564039 TI - In vitro studies of the sensitivity of canine bone-marrow erythroid burst-forming units (BFU-E) and fibroblast colony-forming units (CFU-F) to X-irradiation. AB - The radiosensitivity of the early erythroid progenitor cells (BFU-E) and the progenitor cells of the stroma (CFU-F) in canine bone marrow was studied under steady-state conditions by in vitro irradiation with 280 kV X-rays. The dose effect relationship for colony formation was determined for BFU-E obtained from the iliac crest marrow, and for CFU-F in bone marrow collected from the iliac crest and the humerus of adult beagles. The BFU-E were adequately stimulated with serum from lethally irradiated dogs to obtain a source of BPA (burst-promoting activity). The BFU-E proved to be extremely radiosensitive, and the survival curve was exponential (D0 = 15.3 +/- 1.8 cGy). We showed that buffy-coat leukocytes separated from bone marrow leukocytes obtained by aspiration were an optimum source of CFU-F. A curve was fitted to the data obtained for CFU-F obtained from the iliac crest or the humerus, resulting in D0 = 241 +/- 38 cGy and an extrapolation number n = 1.38 +/- 0.62 or D0 = 261 +/- 40 cGy and n = 1.04 +/- 0.42, respectively. According to these findings, and other published data, we conclude that the canine bone marrow BFU-E are presently the most radiosensitive hemopoietic cells detected among all hemopoietic cells of different mammals. PMID- 2564040 TI - Hematopoietic death of unprotected man from photon irradiations: statistical modeling from animal experiments. AB - Most estimates of the radiation dose lethal to 50 per cent of a human population are based on historical data taken from well-known experiences reflecting inadequately known physical and biological conditions, or from medical procedures where individual patients received advantages of modern clinical care. It has been debated as to whether the experience of unprotected man would more closely reflect that of hospital patients or of the radiobiological studies with large animals. The issue at question is whether the apparent two-fold or more increased susceptibility of large animals to death from bone marrow damage (compared with the majority of estimates for man) is due more to true interspecies differences or the lack of medical support. This study is an attempt to assess the radiosensitivity of unprotected man in terms of the composite animal experiments. Based upon an extensive data base containing 121 separate animals studies using 13 different species, an estimate of the mortality dose-response relationship due to a uniform, continuous field of photon radiation is predicted for 70 kg unprotected man. Man is assumed to have a level of radiation sensitivity similar to that of the species represented in the data base, after adjustment for body weight. The mathematical model used includes fixed terms to account for effects of body weight and dose rate, and random terms reflecting inter- and intra species variation and experimental error. Point predictions and 95 per cent prediction intervals are given for the LD05, LD10, LD25, LD50, LD75, LD90, and LD95, for dose rates ranging from 0.01 to 0.5 Gy/min, and treatment times ranging from about 2 min to about 24 h. At 1 cGy/min our point prediction of the LD50 is 299 cGy with an associated 95 per cent prediction interval of (168 cGy, 535 cGy). The analogous values at 50 cGy/min are 183 cGy and (103 cGy, 326 cGy). PMID- 2564041 TI - Competition for polyamine uptake into rat lung slices by WR2721 and analogues. AB - The objective of these studies was to determine whether a series of structurally related radioprotective agents could act as substrates for the recently identified polyamine system in the lung. We have shown that WR2721 (S-2(3 aminopropylamino)ethyl phosphorothioate), S-2(4-aminobutylamino)ethyl phosphorothioate (S-ABEP or WR2822) and S-2(7-aminoheptylamino)ethyl phosphorothioate (S-AHEP) competitively inhibit the uptake of putrescine into rat lung slices. The ability of the radioprotectors to act as substrates for the polyamine uptake system was expressed as the Ki for each compound. The Ki values for WR2721, S-ABEP and S-AHEP in the absence of dithiothreitol were 48, 57 and 7 mumol dm-3 compared to 155, 88 and 15 mumol dm-3 in the presence of dithiothreitol, indicating that the disulphide form may have a higher affinity for the transport system. By analogy with other substrates for the polyamine uptake system we have concluded that it should be possible to target radioprotectors to the alveolar epithelial type I and II cells and the Clara cells in the lung, as they prossess this uptake system, and thus protect these cells from oxidative stress. PMID- 2564042 TI - Pitfalls in the assessment of late lung damage in irradiated mice: complications related to pleural effusion. PMID- 2564043 TI - Re: Assessment of late lung damage. PMID- 2564044 TI - Radiological heart enlargement in treated hypertensive men: a comparative study of chest X-ray examination and M-mode echocardiography. AB - Twenty-five hypertensives with no history of myocardial infarction and with a radiologically determined heart enlargement were examined with M-mode echocardiography and compared with a normotensive control group (n = 41). All except two of the hypertensive patients were on a beta-blocker based antihypertensive treatment regime. The relative heart volume on X-ray was significantly larger in the hypertensives, 562 ml m-2 body surface area (BSA), compared to the normotensives, 408 ml m-2 BSA (P less than 0.001). Both left ventricular diameter (LVD) in end-diastole and end-systole and left arterial (LA) diameter were significantly larger in the hypertensives (56 vs. 51 mm, P less than 0.01; 35 vs. 31 mm, P less than 0.01; 46 vs. 42 mm, P less than 0.01, respectively) as was the LV mass (296 vs. 203 g, P less than 0.001). The end systolic wall stress (ESWS) was significantly greater in the hypertensives. Despite these findings resting left ventricular fractional shortening was the same and showed a similar correlation with ESWS (r = 0.79 and r = 0.77, respectively) in both groups. Hence, left ventricular systolic performance was not impaired in the hypertensives compared to the normotensives. These results show that an enlarged cardiac silhouette on the chest X-ray in hypertensive subjects with beta-blocker based drug therapy must be interpreted with caution and must not, a priori, be judged as a sign of an impaired systolic cardiac function. PMID- 2564045 TI - Apparent non-involvement of prions in the pathogenesis of spongiform change in HIV-infected brain. AB - Characteristic neuropathological white matter changes in the brains of patients dying from the acquired immune deficiency syndrome (AIDS) suggest pathogenetic involvement of either the PrP27-30 gene or a scrapie-related prion; however, proteinase-K resistant scrapie-like proteins could not be detected in brain tissue from AIDS patients. PMID- 2564046 TI - Mitochondrial storage forms of acetyl CoA carboxylase: mobilization/activation accounts for increased activity of the enzyme in liver of genetically obese Zucker rats. AB - In earlier reports, we have described a previously unrecognized mechanism which regulates the activity of acetyl CoA carboxylase in rat liver by the control of its distribution between relatively inactive mitochondrial and active cytosolic forms. In this study, the activity, total quantity and the subcellular distribution of acetyl CoA carboxylase were determined in liver of fed and fasted (48 h) homozygous obese (fa/fa) zucker rats and homozygous lean (Fa/Fa) littermates. The results indicate that neither diet nor genetic obesity affected the total quantity of acetyl CoA carboxylase per unit weight of liver. Instead, increased activity of this enzyme in the liver of the Zucker rat was primarily due to a shift in the subcellular distribution away from relatively inactive mitochondrial forms toward active cytosolic forms. Thus, the Zucker rat appears to be yet another example illustrating the physiological importance of regulating the activity of acetyl CoA carboxylase by controlling its subcellular distribution. PMID- 2564047 TI - Comparison of dehydroepiandrosterone and clofibric acid treatments in obese Zucker rats. AB - The effects of either dehydroepiandrosterone (DHEA) or clofibric acid (CFA) treatment on obese female Zucker rats were compared. After 5 wk of treatment, food intake, body weight gain and food efficiency ratio of DHEA-treated rats were 85, 64 and 75%, respectively, of those of CFA-treated and control rats. Liver weights of DHEA- and CFA-treated rats were higher than those of control rats. Non fasting serum glucose levels were similar in all groups, but serum insulin level of DHEA-treated rats was 59% of that in CFA-treated and control rats. Results of glucose tolerance tests were not different among the groups. Mitochondrial state 3 and 4 rates expressed per g liver or per liver with either glutamate-malate or succinate as substrate were higher in DHEA-treated rats than in CFA-treated or control rats. Mitochondrial long-chain fatty acyl-CoA hydrolase activity was 8.5 and 4 times higher in DHEA- and CFA-treated rats, respectively, than in control rats. These results suggest that although DHEA and CFA affect some hepatic biochemical parameters similarly, they are distinct in their effects on body weight, serum insulin and mitochondrial respiration in obese Zucker rats. PMID- 2564048 TI - Improved definition of carrier status in X-linked hypohidrotic ectodermal dysplasia by use of restriction fragment length polymorphism-based linkage analysis. AB - The detection of carriers of the X-linked disorder hypohidrotic ectodermal dysplasia is problematic because of random X-inactivation; the diagnosis was previously based on the observation of subtle defects in ectodermal structures in at-risk females. Linkage studies have recently mapped hypohidrotic ectodermal dysplasia to the region Xq11-q21.1. We assessed the improvement in carrier detection by the method of linkage analysis, in which restriction fragment length polymorphisms were used as markers, in 72 at-risk female members of 29 families. Carriers analyses were based on pedigree information, dental examination of at risk females (phenotype), and DNA analyses at seven linked marker loci. Linkage analysis based on restriction fragment length polymorphisms significantly improved risk estimates over those based on phenotype and pedigree alone. When all available information was combined, 85% (61/72) of the at-risk females had final risks of less than 5% or greater than 95%, and 68% (49/72) had risks less than 1% or greater than 99%. A diagnosis of hypohidrotic ectodermal dysplasia was also excluded (97.5% probability) by DNA and linkage analyses from a sample of cord blood from an at-risk male; a similar approach can be taken for prenatal diagnosis of the disorder. PMID- 2564049 TI - Status of dopamine in bovine pineal glands and the stimulation of N acetyltransferase activity by D2-dopaminergic receptor agonists in the rat pineal glands in culture. AB - In a previous study, we identified in the bovine pineal gland two [3H]spiroperidol-binding sites with KD values of 0.18 and 2.1 nM and Bmax values of 37 and 630 fmol/mg protein, respectively. In this study, the status of dopamine in the bovine pineal glands was delineated further by measuring the relative concentrations of dopamine and norepinephrine and the relative concentrations of serotonin and melatonin. Furthermore, the presence of 4.0 +/- 0.6 micrograms/dopamine/gm tissue encouraged us to delineate the effects of select dopaminergic receptor agonists and antagonists on the synthesis of melatonin in vivo and on the activity of N-acetyltransferase in the rat pineal gland in culture. The acute administration of haloperidol (3 mg/kg intraperitoneally [ip]) or sulpiride (200 mg/kg ip) increased the concentration of melatonin in the pineal gland from 160.6 +/- 8.18 to 327.6 +/- 45.43 and 306.5 +/- 40.53 pg/gland, respectively. Dopamine exhibited dual effects on the activity of N-acetyltransferase, inhibiting the basal activity at 0.1 microM and stimulating it at 10 microM, and the later effect was blocked by propranolol. D2 dopaminergic receptor agonists such as bromocriptine (4.0 microM) or LY-171555 (10.0 microM) partially attenuated the norepinephrine-induced stimulation of N acetyltransferase, and these attenuating effects were reversed by D2-dopaminergic antagonists such as haloperidol (10 microM) or domperidone (10 microM). The results of these studies are interpreted to indicate that for the synthesis of melatonin, the pineal D2-dopaminergic receptors may function independently from those of the beta 1-adrenergic receptor sites. Furthermore, the said D2 dopaminergic receptor are amenable to down regulation since the activity of N acetyltransferase remained unaltered (0.0717 vs. 0.0729 nmol/gland/h) following chronic treatment (4 mg/kg ip/day for 30 days) with bromocriptine. PMID- 2564050 TI - Genetic changes in human adrenocortical carcinomas. AB - Recent studies have suggested that loss of heterozygosity at loci on the short arm of human chromosome 11 (11p) may be important in the pathogenesis of benign and malignant adrenal cortical tumors. To test this concept, adrenocortical carcinomas from nine patients and benign adrenal cortical lesions from eight patients were tested for loss of alleles at loci on human chromosomes 11, 13, and 17. All patients with adrenocortical carcinoma whose normal somatic tissues were heterozygous for a locus on chromosome 17p had lost alleles in the tumor. Four of six patients with adrenocortical carcinoma who were heterozygous for one or more alleles on chromosome 11p in normal tissues had lost 11p alleles in the tumor. Three of six patients with adrenocortical carcinoma showed loss of 13q alleles in the tumor. Loss of alleles on chromosomes 11p, 13q, and 17p was observed in primary tumors and metastases but not in adrenocortical adenomas or hyperplastic lesions of the adrenal cortex. One patient with adrenocortical carcinoma had a somatic mutation in the HRAS1 gene in the normal adrenal gland. The consistency of the genetic changes on chromosomes 11p, 13q, and 17p suggests that they are important in the pathogenesis of adrenocortical carcinoma. PMID- 2564051 TI - Transmitter neurochemistry of the efferent neuron system innervating the labyrinth. AB - It is likely that several mechanisms contribute to the efferent control of cochlear and vestibular function. Different effects are probably mediated by different neuronal transmitters. In spite of a number of transmitter candidates, it is still widely assumed that the entire efferent system can be globally characterized as cholinergic. We attempted to label retrogradely identified efferent neurons in the brainstem with a monoclonal antibody against choline acetyltransferase (ChAT), the acetylcholine (ACh) synthesizing enzyme. Only a portion of the vestibular efferents could thus be shown to be cholinergic in the rat. Medial cochlear efferents, terminating under outer hair cells, may also be cholinergic since they stain intensely for acetylcholine esterase (AChE) after pre-treatment with the AChE inhibitor diisopropylfluorophosphate (DFP). The lateral cochlear efferents terminating under inner hair cells, as well as more than half of the vestibular efferent neuron population, reacted negatively with either method designed to identify cholinergic neurons. Half of the lateral olivo cochlear neuron population filled retrogradely with tritiated gamma-amino butyric acid [( 3H]-GABA). These cells were similar in size and distribution to neurons staining for the GABA synthesizing enzyme glutamic acid decarboxylase (GAD). Retrograde transport of [3H]-aspartate from the inner ear to the brainstem was seen in half of the lateral olivocochlear population, as well as in part of the efferent vestibular population in group E and in the caudal pontine reticular nucleus (CPR). Since various peptides have also been located in efferent neurons, this system is chemically diversified. Several distinct mechanisms of efferent control with presumably differing functions must, therefore, exist. PMID- 2564052 TI - [Do benzodiazepines represent "natural" drugs?]. PMID- 2564053 TI - [European Nursing Conference, 21-24 June, 1988 in Vienna. "Vienna Declaration" and recommendations]. PMID- 2564054 TI - Diagnostic value of liver function tests in bile duct obstruction. AB - Serological tests may be of value in differentiating acute and chronic bile duct obstruction because the rate of alteration of hepatic cellular integrity and function will affect the rate of cellular product release. In a canine model the common bile duct was obstructed either suddenly (N = 7) or gradually (N = 5). A control group (N = 5) had the common bile duct dissected free from the surrounding tissues. Blood was taken before and 1, 2, 4, 7, 11, 14, 17, 21, and 28 days after initiating obstruction. Serum alkaline phosphatase, bilirubin, aspartate aminotransferase, alanine aminotransferase, ornithine carbamyl transferase, and gamma-glutamyl transferase levels were significantly greater with sudden compared to gradual occlusion, and the values were larger than those in the control. The range of values of alkaline phosphatase, bilirubin, and aspartate aminotransferase did not overlap in the acute and chronic groups at specific times. Serum albumin and total protein were normal in all groups. The magnitude of alkaline phosphatase, aspartate aminotransferase, and bilirubin elevation may help in the differentiation of acute and chronic biliary obstruction. PMID- 2564055 TI - Effect of calcium entry and beta blockade during infrarenal aortic clamping. AB - Clamping and declamping during aortic surgery produce a hemodynamically significant myocardial stress. The cardiovascular (CV) response to this stress may be adversely altered by calcium antagonists and beta-adrenoreceptor blockade employed to control symptomatic coronary artery disease. This study evaluated the effect of verapamil (V), propranolol (P), and their combination (P + V) on the CV response to infrarenal abdominal aortic cross-clamping and declamping in anesthetized dogs. Six dogs received P as a bolus of 0.5 mg/kg 20 min before clamping. Six additional dogs received V as a 300 micrograms/kg bolus followed by a V infusion of 6 micrograms/kg/min for 20 min before clamping. A third group of six dogs received the P bolus followed 20 min later by the V regimen (P + V). In both the V and P + V groups, 6 micrograms/kg/min V was infused throughout the clamping and declamping sequence. A fourth group of six control dogs received no cardioactive drugs during the experiment. Heart rate, mean aortic blood pressure, left ventricular end-diastolic pressures, peak rate of rise of left ventricular pressure, cardiac output, and systemic vascular resistance were measured in all animals before aortic cross-clamping, at 5 and 40 min after clamping, and 5 min after declamping. The results demonstrated additive negative chronotropic and inotropic properties of P + V therapy with a more significantly adverse effect than that of either drug alone. The implications of this study warrant added caution when patients treated with these drugs undergo abdominal aortic surgery. PMID- 2564056 TI - Gonadal steroids and anterior lobe dynorphin in the male rat. AB - Immunoreactive (ir)-dynorphin levels were measured, and the species characterized by high performance liquid chromatography (HPLC), in the pituitary and hypothalamus of intact and castrate male rats. On HPLC, ir-dynorphin co-eluted with authentic dynorphin A 1-8, dynorphin A 1-17 and dynorphin 1-32 in the hypothalamus and intermediate lobe; in two different reversed phase (RP)-HPLC systems, anterior lobe ir-dynorphin co-eluted uniquely with dynorphin 32 (4K dynorphin). Anterior lobe levels of total ir-dynorphin were significantly lowered 7 days after castration, while HPLC profiles in all tissues remained unchanged. The change in anterior pituitary ir-dynorphin levels was reversed in a dose related manner by dihydrotestosterone (15-500 micrograms/100 g b. wt/day); estradiol benzoate (3 micrograms/100 g/day) was without effect. The changes on castration and androgen administration suggest that gonadal steroids play a role in the regulation of dynorphin, as well as gonadotrophins and prolactin, within the anterior pituitary gland. PMID- 2564057 TI - Patent foramen ovale and decompression sickness in divers. AB - 30 patients with a history of decompression sickness were examined for the presence of patent foramen ovale by bubble contrast, two-dimensional echocardiography and colour flow doppler imaging. With bubble contrast, 11 (37%) of the patients had right-to-left shunting through a patent foramen ovale during spontaneous breathing. 61% of a subset of 18 patients with serious signs and symptoms had shunting. This number was significantly higher than the 5% prevalence seen with the same diagnostic technique in 176 healthy volunteers. The presence of patent foramen ovale seems to be a risk factor for the development of decompression sickness in divers. PMID- 2564058 TI - Rates of thromboembolism with three different mechanical heart valve prostheses: randomised study. AB - In 434 operations for valvular heart disease, patients were randomised to receive Bjork-Shiley, Edwards-Duromedics, or Medtronic-Hall mechanical prostheses. At a median follow-up time of 37.5 months there were no differences in hazard of death or non-embolic events. In the first six postoperative months the incidence of thromboembolism was about the same for the three valves. Subsequently, however, the incidence of thromboembolism for the Edwards-Duromedics valve was 3.9 times higher than for the Bjork-Shiley valve, and for the Medtronic-Hall valve 2.6 times higher than for the Bjork-Shiley valve. PMID- 2564059 TI - Infantile gastroenteritis associated with excretion of pestivirus antigens. AB - Faeces from children under 2 years old who had gastroenteritis that could not be attributed to recognised enteric pathogens were examined with a monoclonal antibody-based immunoassay for Pestivirus antigens. Such antigens were detected in 30 of 128 episodes of gastroenteritis. Children without diarrhoeal disease and children infected with rotaviruses had little evidence of Pestivirus infection (faeces positive in 1 of 28 and 1 of 31, respectively). The diarrhoeal disease in children excreting Pestivirus antigens resembled that in other children except that it was more commonly associated with signs and symptoms of respiratory inflammation. PMID- 2564060 TI - Molecular genetics of amyloid neuropathy in Europe. AB - The Portuguese type of familial amyloid polyneuropathy (FAP type I), a disabling autosomal dominant disorder with onset in early adult life, is caused by a point mutation in the transthyretin (TTR; previously known as prealbumin) gene. DNA analysis in thirteen European families (one British, two French, one Italian, one Greek, and eight Cypriot) showed that members of all those from Cyprus and Greece, and one from France, carried the FAP type I mutation. Patients from seven of these ten kindreds were not known to have a genetic disease before this study, which demonstrated the mutation in 16 of 43 clinically unaffected relatives. 2 of these were aged over 50 years. TTR gene analysis has useful applications in genetic counselling, including prenatal diagnosis, in identifying the cause of seemingly sporadic cases of amyloid neuropathy, and in epidemiological studies of FAP. PMID- 2564061 TI - Transfusions and graft-versus-host disease. PMID- 2564062 TI - Speech dysfluency. PMID- 2564063 TI - Vigabatrin. PMID- 2564064 TI - Auxiliary liver transplantation. PMID- 2564065 TI - A lump in the throat. PMID- 2564066 TI - Effect of methotrexate on relapse after bone-marrow transplantation for acute lymphoblastic leukaemia. International Bone Marrow Transplant Registry. AB - Data from 634 patients who received HLA-identical bone-marrow transplants for acute lymphoblastic leukaemia in first or second remission were analysed to examine the influence of mode of prophylaxis against graft versus host disease on rate of relapse of leukaemia. Methotrexate was associated with a significantly lower risk of leukaemia recurrence than were other methods of GVHD prophylaxis (relative risk 0.2, p less than 0.0003, for first-remission transplants; relative risk 0.3, p less than 0.0001, for second remission transplants). The decreased risk of relapse did not seem to be mediated via an impact on incidence or severity of graft versus host disease. A direct antileukaemia effect of methotrexate is the most likely mechanism. PMID- 2564067 TI - World-wide antibiotic resistance in methicillin-resistant Staphylococcus aureus. AB - Antibiotic resistance patterns were determined for 106 strains of methicillin resistant Staphylococcus aureus (MRSA) from 21 countries. Resistance to gentamicin, tobramycin, netilmicin, amikacin, streptomycin, or erthromycin was recorded in more than 90% of strains. Resistance to the other compounds tested was as follows: tetracycline 86%, minocycline 76%, trimethoprim 69%, clindamycin 66%, neomycin 59%, chloramphenicol 39%, rifampicin 26%, fosfomycin 22%, ciprofloxacin 17%, fusidic acid 12%, bacitracin 2%, and novobiocin 1%. All the stains were sensitive to mupirocin, pristinamycin, ramoplanin, teicoplanin, and vancomycin. There were geographical patterns of resistance: MRSA from the UK and Australia were predominantly resistant to trimethoprim, whereas many strains from centres in Europe and the USA were sensitive. MRSA that were resistant to ciprofloxacin were of French and German origin. 15 strains, 12 of which came from France, Turkey, or Brazil, were resistant either to thirteen or to fourteen agents. PMID- 2564068 TI - How-often-that-high graphs of serum cholesterol. Findings from the Scottish Heart Health and Scottish MONICA studies. AB - The Scottish Heart Health and Scottish MONICA studies included measurements of serum total cholesterol in 10,450 representative men and women aged 25-64 years recruited across Scotland in 1984-86. The results were typical of Britain as a whole. A new graph, called the HOTH graph ("how often that high?"), shows the percentage of the population with serum total cholesterol at or above any given value. Median (equal to mean) cholesterol levels in men were 5.5, 6.0, 6.3, and 6.2 mmol/l in successive 10-year age groups 25-64, whilst equivalent values in women were 5.2, 5.5, 6.4, and 7.2 mmol/l. By comparison with other countries these are high, but the percentage of the population above specific cutpoints is disproportionate, varying considerably by age and sex. Application of imported cholesterol management algorithms, based on global cut-points, would lead to an overwhelming caseload of patients needing intensive lipid investigation and management, dominated by older women, and incurring great costs. 35% of the population 25-64 years old would be at or above 6.5 mmol/l and 11% at or above 7.8 mmol/l. A population diet and multiple-risk-factor strategy would be more feasible and rational than a one-dimensional cholesterol cut-point approach. PMID- 2564069 TI - Conservative management of an echovirus 11 outbreak in a neonatal unit. AB - Two babies in a neonatal unit presented on the same day with meningitis due to echovirus 11, which was thought to have been introduced by staff. At this time echovirus 11 was also isolated from the stools of eight other babies; five of them did not have signs of infection. No intervention was made except to emphasise the importance of handwashing. There was evidence of secondary spread to two babies who were both clinically well. The attack rate was twelve (29%) of forty-one babies exposed. Seven of the twelve infected babies were born before 30 weeks' gestation and would have had little or no maternal antibody, yet only two of the seven babies had signs of infection. Despite lack of special measures, all babies recovered. Most cases of horizontally acquired neonatal echovirus infection are mild: extreme measures in the management of outbreaks are unnecessary. PMID- 2564070 TI - Working Paper 6 on the National Health Service. PMID- 2564071 TI - Embryo biopsy. PMID- 2564072 TI - Population screening for serum cholesterol. PMID- 2564073 TI - Self-control training in blepharospasm. PMID- 2564074 TI - Listeriosis and ready-cooked chicken. PMID- 2564075 TI - Vitamin D and breast cancer. PMID- 2564076 TI - Resistant hypertension due to pica (baking soda) PMID- 2564077 TI - Immune dysfunction in patients not responding to hepatitis B vaccination. PMID- 2564078 TI - Oral contraceptives and breast cancer: is the CASH study really negative? PMID- 2564079 TI - Duplex Doppler analysis of renal vasculature. PMID- 2564080 TI - HIV-2 in West Africa. PMID- 2564081 TI - Peripheral facial nerve palsy associated with HIV infection. PMID- 2564082 TI - Electroencephalography and HIV infection. PMID- 2564083 TI - Identification of EBV-DNA in tumour cells of AIDS-related lymphomas by in-situ hybridisation. PMID- 2564084 TI - Debrisoquine hydroxylation polymorphism and personality. PMID- 2564085 TI - Unusual transmission of falciparum malaria in Italy. PMID- 2564086 TI - Cetirizine in pollen-associated asthma. PMID- 2564087 TI - Prostaglandins in irradiation-induced mucositis. PMID- 2564088 TI - HTLV-I positive spastic paraparesis in a temperate zone. PMID- 2564089 TI - Maternal mortality in a Sahelian capital. PMID- 2564090 TI - NHS white-paper. PMID- 2564091 TI - Clinical trials in the United States. PMID- 2564092 TI - Kidneys for sale. PMID- 2564093 TI - Interleukin-2 receptor levels indicating relapse in multiple sclerosis. PMID- 2564094 TI - Isothiazolinone. PMID- 2564095 TI - Left-ventricular support by intraventricular blood pump during high-risk coronary angioplasty. PMID- 2564096 TI - When laser vaporisation for CIN fails, what next? PMID- 2564097 TI - Correcting total serum calcium. PMID- 2564098 TI - Asthma and rhinitis associated with lycopodium spores on condoms. PMID- 2564099 TI - Corticosteroid-induced osteoporosis and hormone implants. PMID- 2564100 TI - Plastic or paraffin? PMID- 2564101 TI - Anaerobic curved rods in breast abscess and vagina. PMID- 2564102 TI - Diet and coronary risk in men and women. PMID- 2564103 TI - Symptomatic relief of meningioma by buserelin maintenance therapy. PMID- 2564104 TI - Aluminium in infant formulas. PMID- 2564105 TI - Fetal brain transplantation for Parkinson's disease: technique for obtaining donor tissue. PMID- 2564106 TI - Side-effects of subcutaneous apomorphine in Parkinson's disease. PMID- 2564107 TI - Undifferentiated cells, metaplasia, c-myc expression, and gastrointestinal carcinogenesis. PMID- 2564108 TI - Paracetamol overdose: timing the antidote. PMID- 2564109 TI - Adjuvant chemotherapy in operable gastric cancer. 5 year follow-up of first British Stomach Cancer Group trial. AB - 411 patients were entered into a prospective, randomised controlled trial of adjuvant chemotherapy after gastrectomy for adenocarcinoma. After a follow-up of at least 5 1/2 years there has been no survival advantage for those receiving adjuvant 5-fluorouracil and mitomycin C with or without an induction course of 5 fluorouracil, vincristine, cyclophosphamide, and methotrexate compared with those undergoing surgery only. There have been 366 deaths, including 22 from treatment related conditions. A multivariate analysis of prognostic factors demonstrated that stage of disease, nodal and resection margin involvement, and the presence of residual disease are significant determinants of survival. Weight loss before surgery had a significant independent influence on survival. The combination of preoperative symptoms and intraoperative findings may be used to select patients for radical or palliative procedures. PMID- 2564110 TI - Comparison of rifampicin with phenobarbitone for treatment of pruritus in biliary cirrhosis. AB - The anti-pruritic effects of rifampicin (10 mg/kg) and phenobarbitone (3 mg/kg) were assessed in 22 patients with primary biliary cirrhosis in a crossover randomised clinical trial. Each agent was given for 14 days, with a 30-day washout period between treatments. 21 patients completed the course of rifampicin and 18 that of phenobarbitone; rifampicin was withdrawn from 1 patient when anaemia and renal failure developed, whereas 3 patients stopped taking phenobarbitone because of a rash and the 4th merely refused the drug. Rifampicin had a greater anti-pruritic effect than phenobarbitone. The symptom improved in 19 patients taking rifampicin and in 8 taking phenobarbitone, the degree of improvement being greater with rifampicin than with phenobarbitone. Pruritus disappeared in 9 patients receiving rifampicin, and three of them were free of itch when switching over to phenobarbitone. Both drugs were equally effective in inducing hepatic microsomal function but rifampicin has the additional effect of reducing cholestasis. Its anti-pruritic effect should be tested in long-term clinical trials. PMID- 2564111 TI - Tumour-infiltrating lymphocytes and interleukin-2 in treatment of advanced cancer. AB - Tumour-infiltrating lymphocytes (TIL) were isolated and expanded from small tumour biopsy samples of twenty-eight patients (thirteen with malignant melanoma, seven with renal cell carcinoma, and eight with non-small-cell lung cancer). The patients were treated with autologous expanded TIL (about 10(10)) and continuous infusions of recombinant human interleukin-2(1-3 x 10(6) U/m2 per 24 h). 29% of the patients with renal cell cancer and 23% of those with melanoma achieved objective tumour responses lasting 3-14 months. Toxic side-effects were limited, and no patient required intensive-care monitoring. Adoptive immunotherapy with TIL and interleukin-2 may be an effective systemic approach to the treatment of some patients with malignant melanoma and renal cell carcinoma. PMID- 2564112 TI - Field testing and comparative evaluation of rapid, visually read screening assays for antibody to human immunodeficiency virus. AB - Five rapid, visually read assays for detection of antibody against human immunodeficiency virus (HIV) were evaluated on fresh serum samples from 4000 prospective blood donors at Mama Yemo Hospital, Kinshasa, Zaire. The sensitivity of the assays, based on 214 specimens positive by western blot, ranged from 84.6% to 99.1%. The specificity, based on 3664 samples negative by enzyme-linked immunosorbent assay (ELISA) or western blot, ranged from 92.7% to 98.8%. Three readers scored each test result independently; disagreement about test interpretation occurred in 1.2-8.3% of the specimens. There was no correlation between assay performance and assay principle (agglutination or dot immunobinding) or antigen source (viral lysate or recombinant). Assays such as these can be readily implemented in a developing country transfusion centre, where blood screening by ELISA is not practicable. PMID- 2564113 TI - Urinary leukotriene E4 after antigen challenge and in acute asthma and allergic rhinitis. AB - The leukotrienes LTC4, D4, and E4 are potent bronchoconstrictor agents and are thought to have an important role in asthma. Urinary LTE4, a stable urinary end product of LTC4 and LTD4, was measured, by means of high-performance liquid chromatography and radioimmunoassay. LTE4 excretion followed a log-normal distribution in twenty-nine healthy controls, with a geometric mean of 23.8 (95% confidence interval 19.9-28.2) ng/mmol creatinine. Urine was collected from eight atopic subjects for 3 h after antigen inhalation and a control urine collection was made a week later at the same time of day. Urinary LTE4 was significantly higher after antigen challenge than in the control sample (153.7 [87.1-271.3] vs 23.5 [13.7-69.5] ng/mmol creatinine; p less than 0.01). Urinary LTE4 was also measured in twenty patients with severe acute asthma and nine patients with seasonal allergic rhinitis. Mean urinary LTE4 was higher in the asthmatic patients (78.3 [46.5-131.8] ng/mmol creatinine) than in normal subjects (p less than 0.01), although there was substantial overlap into the normal range. The urinary LTE4 values of the rhinitis patients were within the normal range whether or not they had symptoms. LTC4 and LTD4 were also found in bronchoalveolar lavage fluid from one of the three atopic subjects challenged with antigen before lavage, and in a single patient who underwent lavage after admission with severe acute asthma. These studies provide evidence that leukotrienes are released in vivo in man after antigen challenge and in acute asthma. PMID- 2564114 TI - Type 2 diabetes or NIDDM: looking for a better name. PMID- 2564115 TI - Is the Apgar score outmoded? PMID- 2564116 TI - PAF antagonists in asthma. PMID- 2564117 TI - Routine arthroscopy for acute haemarthrosis of the knee. PMID- 2564118 TI - The journal club--let's audit. PMID- 2564119 TI - Factors influencing mortality after curative resection for large bowel cancer in elderly patients. AB - Mortality rates from the Large Bowel Cancer Project are presented with special reference to patients older than 70 years. The in-hospital mortality rate among those who underwent curative resection for colorectal carcinoma was 7%. Unlike long-term prognosis, which is influenced by pathological features, in-hospital mortality is influenced largely by clinical factors. Age was an adverse factor (78% of deaths occurred among those aged over 70, who formed 46% of the study population), as was obstruction or perforation. 55% of deaths were due to cardiopulmonary complications. Educating patients to seek treatment early, careful preoperative assessment and postoperative monitoring of cardiopulmonary function, and, in selected patients, use of local treatments rather than wide resections may help to reduce mortality in elderly patients. PMID- 2564120 TI - The prepuce: a mistake of nature? AB - Retrospective studies suggest that circumcision of newborn boys will reduce the frequency of male early infantile urinary tract infection (UTI) by about 90%. If they are correct, this will be the first known instance of a common potentially lethal disease being preventable by extirpation of a piece of normal tissue. To reconcile the phenomenon with existing views of evolution and biology, it is suggested that the effects of one unphysiological intervention are counterbalancing those of another--ie, colonisation of the baby's gastrointestinal tract and genitals in maternity units by Escherichia coli strains of non-maternal origin, to which the baby has no passive immunity. As an alternative to circumcision to prevent early infantile male UTI, more natural colonisation could be promoted by strict rooming-in of mother and baby or by active colonisation of the baby with his mother's anaerobic gut flora. PMID- 2564121 TI - Importance of the clicking hip in screening for congenital dislocation of the hip. AB - A birth cohort of 3289 babies was investigated prospectively for one year to assess the importance of risk factors for congenital dislocation of the hip (CDH). 426 (13%) babies had an identifiable risk factor when examined by paediatric senior house officers. On further examination in the first week of life at a screening clinic, 51 proved to have CDH, an incidence of 15.5 per 1000 births. The results confirmed the importance of established factors such as family history, caesarean section, and the breech position, and demonstrated the clinical significance of the presence of clicking hips on initial examination. After up to 4.1 years of follow-up there have been no late presentations. By taking into account the clinical examination by senior house officers, secondary screening of babies at high risk of CDH can be very effective. This extended high risk group may contain all potentially abnormal hips. All such babies should be examined by ultrasonography in hospitals where it is available. PMID- 2564123 TI - Medical negligence and no-fault liability. PMID- 2564122 TI - Primary cutaneous bacillus cereus infection in neutropenic children. AB - A review of culture reports for a five-year period at St Jude Children's Research Hospital yielded 10 cases of primary cutaneous Bacillus cereus infection in neutropenic patients treated for cancer or aplastic anaemia. Vesicles or pustules were seen only on the limbs. The infections, all of which arose in the spring or summer, responded to antibiotics. In neutropenic patients B cereus should thus be regarded as a possible cause of isolated vesicles. PMID- 2564124 TI - Called to account. A radiologist. PMID- 2564125 TI - Haemorrhoids, constipation, and hypertensive anal cushions. PMID- 2564126 TI - Inhibition of epoxide hydrolase by anticonvulsants and risk of teratogenicity. PMID- 2564127 TI - Valproate and spina bifida. PMID- 2564128 TI - "Leukoaraiosis" explained. PMID- 2564129 TI - Latitude and ischaemic heart disease. PMID- 2564130 TI - Safety of divers. PMID- 2564131 TI - Nephrotic syndrome after injections of bovine cartilage and marrow extract. PMID- 2564132 TI - Aluminium and infant formulae. PMID- 2564133 TI - Defloration as risk factor for heterosexual HIV transmission. PMID- 2564134 TI - Oral contraceptives and breast cancer. PMID- 2564135 TI - Adjuvant tamoxifen and second cancers. PMID- 2564136 TI - Cervical cancer deaths in young women. PMID- 2564137 TI - Listeria in hospital lettuce. PMID- 2564138 TI - Butane sniffing causing ventricular fibrillation. PMID- 2564139 TI - Diltiazem and heart block. PMID- 2564140 TI - Drug advertising. PMID- 2564141 TI - Fourth European meeting on hypertension. PMID- 2564142 TI - Mental tests on elderly patients. PMID- 2564143 TI - Menkes' disease: a disorder of zinc metabolism? PMID- 2564144 TI - 99mTc-HMPAO washout in prognosis of stroke. PMID- 2564145 TI - Cerebral malaria, missed again. PMID- 2564146 TI - Is fine-needle aspiration of tumours harmless? PMID- 2564147 TI - Rice water/salt solution for diarrhoea. PMID- 2564148 TI - Anti-cardiolipin antibodies in viral infection. PMID- 2564149 TI - Case for midline incisions. PMID- 2564150 TI - Hypnotherapy in irritable bowel syndrome. PMID- 2564151 TI - Surgery for gastric cancer. PMID- 2564152 TI - Binding-defective low-density lipoprotein in family with hypercholesterolaemia. PMID- 2564153 TI - Carbamazepine for cocaine addiction? PMID- 2564154 TI - Rectal and colonic varices in cirrhosis. PMID- 2564155 TI - Erythropoietin deficiency in acute renal failure. PMID- 2564156 TI - Intradermal hepatitis B vaccine. PMID- 2564157 TI - Vancomycin and nephrotoxicity. PMID- 2564158 TI - Life-threatening Salmonella enteritidis phage type 4 gastroenteritis in infancy. PMID- 2564159 TI - AIDS and life assurance. PMID- 2564160 TI - Phylogenetic analysis using insertion sequence fingerprinting in Escherichia coli. AB - Chromosomal DNA from 23 closely related, pathogenic strains of Escherichia coli was digested and probed for the insertion sequences IS1, IS2, IS4, IS5, and IS30. Under the assumption that elements residing in DNA restriction fragments of the same apparent length are identical by descent, parsimony analysis of these characters yielded a unique phylogenetic tree. This analysis not only distinguished among bacterial strains that were otherwise identical in their biochemical characteristics and enzyme electrophoretic mobilities, but certain aspects of the topology of the tree were consistent across several unrelated insertion elements. The distribution of IS elements was then reexamined in light of the inferred phylogenetic relationships to investigate the biological properties of the elements, such as rates of insertion and deletion, and to discover apparent recombinational events. The analysis shows that the pattern of distribution of insertion elements in the bacterial genome is sufficiently stable for epidemiological studies. Although the rate of recombination by conjugation has been postulated to be low, at least two such events appear to have taken place. PMID- 2564161 TI - [Benzodiazepines in pregnancy]. PMID- 2564162 TI - Hereditary ataxia. AB - The hereditary ataxias, also referred to as the spinocerebellar degenerations, comprise a series of clinical manifestations that include ataxia and dysmetria, resulting from the predominant involvement of the cerebellum and its afferent and efferent pathways. These disorders are system degenerations; many of them are specific entities clearly inherited as autosomal dominant or autosomal recessive traits. Although the clinical manifestations and neuropathologic findings of cerebellar disease dominate the spinocerebellar degenerations, there may also be characteristic changes in the basal ganglia, optic atrophy, retinitis pigmentosa, or peripheral nerve disease. There are many gradations from pure cerebellar manifestations to mixed cerebellar and brain-stem disorders, cerebellar and basal ganglia syndromes, and spinal syndromes or peripheral nerve disease. The clinical picture may be consistent in one family, but sometimes there is a characteristic syndrome in the majority of family members and an entirely different disorder in one or several members. PMID- 2564163 TI - [Antihistamines in pediatrics]. AB - Histamine antagonists have various effects on H1, H2, cholinergic, adrenergic and serotoninergic receptors. In childhood side effects may be different and central agitation may be more pronounced than in adults. Histamine antagonists should be avoided in the fetal and neonatal periods. They are not indicated in common cold. The use of histamine antagonists in childhood will be discussed. PMID- 2564164 TI - Drug therapy. Combined beta-adrenergic and calcium-entry blockade in angina pectoris. PMID- 2564165 TI - How Escherichia coli infects the urinary tract. PMID- 2564166 TI - Prions. Sheep disease in human clothing. PMID- 2564167 TI - Transcripts of one of two Drosophila cyclin genes become localized in pole cells during embryogenesis. AB - Cyclins, originally discovered in the eggs of marine invertebrates, are proteins which undergo dramatic cycles of synthesis followed by degradation at the metaphase-anaphase transition of cell division. That they participate in the G2-M transition is supported by the fact that when synthetic cyclin messenger RNAs from clam and sea urchin are microinjected into the G2-arrested oocytes of Xenopus, they induce maturation. The cyclin of fission yeast is the product of the cdc13 gene, which is known to interact with cdc2, a gene required for the entry into mitosis. We have cloned the genes that encode A-type and B-type cyclins from Drosophila melanogaster by virtue of their sequence similarity to oligonucleotides corresponding to conserved regions of the cyclin genes. We show that both genes encode abundant maternal mRNAs, but whereas the cyclin A mRNA is relatively uniformly distributed before cell formation, the cyclin B mRNA becomes localized to the developing pole cells. In larvae, cyclin A is expressed predominantly in brain and imaginal disks, whereas cyclin B transcripts are abundant in testes. PMID- 2564168 TI - Linkage of a prion protein missense variant to Gerstmann-Straussler syndrome. AB - Gerstmann-Straussler syndrome is a rare familial neurodegenerative condition that is vertically transmitted, in an apparently autosomal dominant way. It can also be horizontally transmitted to non-human primates and rodents through intracerebral inoculation of brain homogenates from patients with the disease. The exact incidence of the syndrome is unknown but is estimated to be between one and ten per hundred million. Patients initially suffer from ataxia or dementia and deteriorate until they die, in one to ten years. Protease-resistant prion protein (PrP) and PrP-immunoreactive amyloid plaques with characteristic morphology accumulate in the brains of these patients. Current diagnostic criteria for Gerstmann-Straussler syndrome incorporate clinical and neuropathological features, as animal transmission studies can be unreliable. PrP is implicated in the pathogenesis and transmission of the condition and in scrapie, an equivalent animal disease. It was discovered by enriching scrapie infected hamster brain fractions for infectivity. Because there is compelling evidence that the scrapie isoform of PrP is a necessary component of the infectious particle, it seemed possible that the PrP gene on the short arm of human chromosome 20 in Gerstmann-Straussler syndrome might be abnormal. We show here that PrP codon 102 is linked to the putative gene for the syndrome in two pedigrees, providing the best evidence to date that this familial condition is inherited despite also being infectious, and that substitution of leucine for proline at PrP codon 102 may lead to the development of Gerstmann-Straussler syndrome. PMID- 2564169 TI - Somatostatin and beta-endorphin levels in cerebrospinal fluid of nonmedicated and medicated patients with epileptic seizures. AB - Neuropeptides have been proposed to play a role in regulation of the seizure threshold and interictal behavior in experimental models of epilepsy, but there are few studies concerning neuropeptides in human epilepsy. We compared the levels of two peptides, somatostatin (SLI) and beta-endorphin (BEP) in lumbar cerebrospinal fluid (CSF) of unmedicated (N = 18) and medicated (n = 24) epileptic patients with the levels of these peptides in control (n = 20). Peptide levels in the CSF of patients with panic disorder (8) were also evaluated. Patients with chronic medicated epilepsy had a SLl level 80% (p = 0.003, Mann Whitney U-test) that of the controls, 76% (p = 0.011) that of unmedicated patients, and 84% (p = 0.028) that of the panic group. BEP in the CSF did not differ in unmedicated, medicated and control patients. On the other hand, patients with panic disorder had higher levels of BEP in CSF than did the controls (117%, p = 0.041). In panic patients SLl was at control level. The present study indicates that the peptidergic systems are affected differentially in epilepsy and in panic disorder. Furthermore, there seems to be selectivity in the affect on peptidergic systems during the period when the epilepsy becomes chronic. PMID- 2564170 TI - Activation of the bilateral corticostriatal glutamatergic projection by infusion of GABA into thalamic motor nuclei in the cat: an in vivo release study. AB - The unilateral application of GABA (10(-5) M; 30 min) into thalamic motor nuclei of the cat increases the release of dopamine in both caudate nuclei. This effect has been suggested to be related to an activation of the bilateral corticostriatal glutamatergic projection, glutamate exerting a presynaptic facilitatory influence on dopamine release. To explore this hypothesis further, halothane-anesthetized cats implanted with push-pull cannulae were used in order to examine the effects of such a GABA application on the release of glutamate in both caudate nuclei. Aspartate, alanine, glutamine, serine and tyrosine were also measured in the superfusates. The unilateral application of GABA (10(-5) M; 30 min) into thalamic motor nuclei increased the release of glutamate bilaterally. Although less pronounced, ipsi- or bilateral increases in the efflux of alanine, glutamine and tyrosine were also observed. Contralateral changes in the efflux of glutamate, alanine and tyrosine were prevented following acute section of the corpus callosum. In addition, when applied continuously into one caudate nucleus, 2-amino-5-phosphonovaleric acid, a blocker of N-methyl-D-aspartate receptors, prevented the GABA-induced increase in alanine or tyrosine efflux but did not affect the enhanced release of glutamate. These results confirm that the unilateral application of GABA in thalamic motor nuclei activates a thalamo cortico-striatal neuronal loop leading to the stimulation of glutamate release in both caudate nuclei. Changes in the efflux of other amino acids could be linked to increased metabolic activity of striatal target cells resulting from the increased release of glutamate and from its effect on N-methyl-D-aspartate receptors. PMID- 2564171 TI - Linear relation between the magnitude of long-term potentiation in the dentate gyrus and associative learning in the rat. A demonstration using commissural inhibition and local infusion of an N-methyl-D-aspartate receptor antagonist. AB - Field potentials were recorded in the dentate gyrus of freely-moving rats in a classical conditioning paradigm in which high-frequency stimulation of the perforant path served as a conditioned stimulus. Paired or unpaired perforant path stimulus-footshock presentations were given to animals engaged in a previously acquired food-motivated lever-pressing task. Conditioned suppression of lever-pressing was the behavioural measure of conditioning. Perforant path stimulus trains at an intensity above spike threshold induced long-term potentiation of synaptic transmission in the dentate gyrus. In this condition, animals learned the perforant path stimulus-shock association. Three strategies were employed to block the induction or reduce the magnitude of long-term potentiation induced by the conditioned stimulus: (1) reduction of the intensity of the stimulus below the spike threshold resulted in no long-term potentiation and a failure by the animals to learn the perforant path stimulus-shock association; (2) inhibitory modulation of long-term potentiation by high frequency activation of commissural input to the dentate gyrus resulted in learning deficits; (3) chronic infusion of DL-2-amino-5-phosphonovalerate, a selective antagonist of the N-methyl-D-aspartate subtype of glutamate receptor, blocked the induction of long-term potentiation and prevented associative learning. A highly significant linear relation emerged from a correlational analysis between the magnitude of the change in synaptic efficacy at the activated synapses and the amount the animals learned about the perforant path stimulus-shock association. The results presented in this paper are consistent with the hypothesis that associative learning depends on the development of lasting changes in synaptic function. We propose that the activation of N-methyl D-aspartate receptors in the dentate gyrus is involved in this process and that the more change in synaptic efficacy is produced in the activated network, the more the animals learn. PMID- 2564172 TI - Long-term potentiation in the hippocampus of the anaesthetized rat is not associated with a sustained enhanced release of endogenous excitatory amino acids. AB - The relationship between long-term potentiation of synaptic transmission and the release of endogenous glutamate and aspartate has been investigated in the CA1 region of the hippocampus and in the fascia dentata of the anaesthetized rat. A high-frequency train of electrical stimulation of afferent pathways produced a long lasting (greater than 2 h) enhancement of the field excitatory postsynaptic potential in CA1 and of the population spike in the fascia dentata. In both regions, this was not associated with a significant long lasting increase in the release of glutamate and aspartate. It is concluded that the maintenance of long term potentiation is not associated with a sustained increase in the release of excitatory amino acids. PMID- 2564173 TI - Repeated administration of SCH 23390 enhances the SKF 38393-induced inhibition in the rat hippocampus. AB - The functional modification of the D1 dopamine receptor subtype following acute or repeated administration of the D1 receptor antagonist SCH 23390 (0.5 mg/kg s.c.) was studied in the rat hippocampal slice preparation. The activity of the D1 receptor system was evaluated by measuring the effect of the D1 receptor agonist SKF 38393 on the spontaneous firing of CA1 hippocampal neurons. The testing was performed 1, 2 and 7 days after discontinuation of the treatment. Repeated (21 days, once daily), but not acute, administration of SCH 23390 significantly potentiated the inhibitory reaction to SKF 38393. The inhibition evoked by SKF 38393 was blocked by application of SCH 23390 (10(-8) M). The results show that repeated treatment with SCH 23390 enhances the inhibitory effect of SKF 38393 in the rat hippocampus, probably due to an increase in the number of D1 dopamine receptors. PMID- 2564174 TI - Characterization of excitatory amino acid receptors in cultured chick cerebellar neurons. AB - In order to characterize excitatory amino acid receptors in cultured chick cerebellar neurons, the effects of amino acid agonists on the input resistance and the antagonist specificity of the depolarization induced by each agonist were intracellularly studied. In Mg-containing medium, glutamate (especially at low doses), aspartate and N-methyl-D-aspartate not only decreased the input resistance at depolarized membrane potentials but also increased it at around the resting potential. In Mg-free medium, glutamate (high and low doses) and all other agonists simply decreased the input resistance. The effects of antagonists on amino acid-induced depolarizations in Mg-free medium were as follows: Mg and alpha-aminoadipate blocked N-methyl-D-aspartate and aspartate most strongly, glutamate and kainate moderately, and quisqualate only slightly; 2-amino-4 phosphonobutyrate antagonized N-methyl-D-aspartate most strongly, aspartate moderately and others mildly; 2-amino-5-phosphonovalerate blocked aspartate most strongly and others mildly; gamma-D-glutamylglycine blocked kainate most strongly and others moderately; and kynurenate was rather nonselective but most strongly antagonized N-methyl-D-aspartate and aspartate. These results suggest that all receptor subtypes (N-methyl-D-aspartate-, quisqualate- and kainate-types) are present in cultured chick cerebellar neurons, but their antagonist specificities are different from those in other central neurons, and also that glutamate at a low dose activates N-methyl-D-aspartate receptors, while it acts on quisqualate receptors at a high dose. PMID- 2564175 TI - Klebsiella pneumoniae infection complicating a puncture wound of the foot: a case report. AB - Klebsiella pneumoniae infections of the feet are rare following puncture wounds. We present a case of such an infection following a nail injury, and stress that there is nothing distinctive about the clinical presentation with this organism and that bacterial cultures and sensitivity tests of isolates are necessary for proper wound management. PMID- 2564176 TI - Effects of bethanechol and adreno-blockers on thermoregulation in spinal cord injury. AB - The authors report two cases of hypothermia due to a treatment associating Bethanechol and Adreno-Blockers. They emphasize the mechanisms of thermoregulation and discuss the pathophysiology of such hypothermia incidents. The most evident explanation is heat loss, principally mediated through Bethanechol, whereas mechanisms of heat preservation are prevented by Adreno Blockers. Patients susceptible to this risk must be carefully monitored. PMID- 2564177 TI - Bacterial exotoxins: how they work. PMID- 2564178 TI - The Mycobacterium tuberculosis BCG-a protein has homology with the Escherichia coli GroES protein. PMID- 2564179 TI - The nucleotide sequence of the fanD gene encoding the large outer membrane protein involved in the biosynthesis of K99 fimbriae. PMID- 2564180 TI - A TaqI RFLP of the human T-cell receptor gamma (TCRG) variable gene V11. PMID- 2564181 TI - SacI polymorphism defined by human T-cell receptor gamma (TCRG) variable gene V11. PMID- 2564182 TI - A rare c-mos RFLP in normal (noncancerous) fibroblasts from two related cancer patients. PMID- 2564183 TI - An NcoI RFLP associated with the gene encoding the alpha-3 chain of human type VI collagen (COL6A3). PMID- 2564184 TI - Two KpnI restriction fragment alleles of the human T-cell receptor delta (TRD) joining segment J2. PMID- 2564185 TI - A new TaqI allele at DXS52 frequent in Algeria. PMID- 2564186 TI - Human insulin receptor RFLPs detected by HindIII and DraI. PMID- 2564187 TI - A polymorphic locus [D1S88] is detected by probe LA01.41 on chromosome 1p. PMID- 2564188 TI - A probe from an X-Y homology region detects RFLPs in Xq13-q22. PMID- 2564190 TI - Additional polymorphism for D7S8 linked to cystic fibrosis including detection by DNA amplification. PMID- 2564189 TI - A stable core region of the tra operon mRNA of plasmid R1-19. AB - The degradation of the polycistronic tra-mRNA of the resistance plasmid R1-19 leads to the accumulation of a well defined series of stable mRNA species. The majority of the most stable mRNAs contains the message for the traA gene only. The differently sized stable mRNAs possess a common 3'terminus within the traL gene but vary at their 5' ends. The 3'terminus probably results from protection against exoribonucleases by a secondary structural feature. We propose that the 5' ends are generated by endoribonucleolytic cleavage. The stability of this part of the tra-mRNA exceeds 30 minutes and probably increases the rate of expression of the traA gene product propilin, the precursor of the sex pilus subunit. The expression of propilin and its processing into a protein of the molecular weight of mature pilin is demonstrated with the isolated gene. The sequence of the so far unknown genes traL and traE of R1-19 is presented. PMID- 2564191 TI - RFLPs of the gene for the human glycine receptor on the X-chromosome. PMID- 2564192 TI - A StuI RFLP upstream of the low density lipoprotein receptor (LDLR) gene. PMID- 2564193 TI - The anonymous DNA probe p7-26 identifying the locus [D7S17], reveals an XmnI polymorphism. PMID- 2564194 TI - RFLP identified by the anonymous DNA segment FZ IV E11 at 1p23 [HGM9 no.D1S87]. PMID- 2564195 TI - RFLP identified by the anonymous DNA segment OLVIIF5 at 21q22.1-22.3 [HGM9 no.D21S143]. PMID- 2564196 TI - A highly polymorphic locus on chromosome 11 which has homology to a collagen triple-helix coding sequence. PMID- 2564197 TI - A pitfall in the prenatal diagnosis of beta-thalassaemia by RFLP. PMID- 2564198 TI - [Occurrence of germ cells in a cryptorchism testis in the adult]. PMID- 2564199 TI - [Anaphylactic shock in patients treated with beta-blockaders]. PMID- 2564200 TI - [Anti-cytoplasmic antibodies of neutrophil granulocytes in systemic vasculitis]. AB - Autoantibodies to the cytoplasmic components of neutrophil polymorphonuclear granulocytes (ACPA) were searched for by indirect immunofluorescence on ethanol fixed smears from normal white blood cells in 248 sera from patients with Wegener's granulomatosis, Kawasaki disease and various other vasculitides, connective tissue diseases, nephropathies and infiltrative pulmonary diseases. Apart from a weak positivity in a few Kawasaki disease sera, ACPA were found in Wegener's granulomatosis (and micropolyarteritis) only, with a correlation between ACPA titers and disease activity. These results confirm the value of ACPA in the diagnosis and evaluation of activity of Wegener's granulomatosis. PMID- 2564201 TI - [Drug surveillance of beta-blockers in eyedrops]. PMID- 2564202 TI - Increased capacity for glutathione synthesis enhances resistance to radiation in Escherichia coli: a possible model for mammalian cell protection. AB - A strain of Escherichia coli, enriched in its content of gamma-glutamylcysteine synthetase and glutathione synthetase activities by recombinant DNA techniques, is more resistant to the lethal effects of gamma-irradiation than is the corresponding wild strain. Although the gene-enriched strain has higher glutathione levels than the wild strain, the observed radioresistance appears to be associated with the increased capacity of the gene-enriched strain to synthesize glutathione when irradiated rather than to the cellular levels of glutathione per se. Thus, resistance was abolished in the presence of buthionine sulfoximine, a selective inactivator of gamma-glutamylcysteine synthetase that decreases glutathione synthesis but that does not act directly to lower cellular glutathione levels. Conclusions drawn from studies on this E. coli model system may have relevance to protection of mammalian cells by glutathione. PMID- 2564203 TI - Purification of a putative brain somatostatin receptor. AB - The brain somatostatin (somatotropin release-inhibiting factor; SRIF) receptor was purified by affinity chromatographic techniques. A protein of 60 kDa could be purified from rat brain. The protein was eluted from a [D-Trp8]SRIF affinity column with either sodium acetate (pH 5.5) or free [D-Trp8]SRIF. The binding of the protein to the affinity column was prevented by free [D-Trp8]SRIF or the stable SRIF analogue SMS 201-996 but not by the inactive somatostatin 28-(1-14). The purified receptor could be covalently labeled by the 125I-labeled SRIF analogue CGP 23996. Excess [D-Trp8]SRIF blocked the binding of 125I-labeled CGP 23996 to the purified receptor, but somatostatin 28-(1-14) did not affect the binding. A 60-kDa protein was also purified from the anterior pituitary cell line AtT-20, which has a high expression of SRIF receptors. In contrast, no 60-kDa protein could be purified from CHO cells, which have no detectable SRIF receptors. These findings present evidence for the purification of the SRIF receptor. PMID- 2564204 TI - Programmed cell death (apoptosis) in pancreatic cancers of hamsters after treatment with analogs of both luteinizing hormone-releasing hormone and somatostatin. AB - Female Syrian golden hamsters with N-nitrosobis(2-oxopropyl)amine (BOP)-induced ductal pancreatic cancers were treated with long-acting microcapsular preparations of the 6-D-tryptophan analog of luteinizing hormone-releasing hormone [( D-Trp6]LH-RH), releasing 25 micrograms/day; the somatostatin analog D Phe-Cys-Tyr-D-Trp-Lys-Val-Cys-Trp-NH2 (RC-160), liberating 15 micrograms/day; and the combination of these two peptides. Therapy with analogs was initiated 24 weeks after initial administration of BOP. These treatments resulted in significantly better survival of all animals as compared to BOP controls; body weights of surviving peptide-treated animals were significantly higher than those of the BOP controls. All 15 BOP-control animals had pancreatic cancers. In the group treated with RC-160 four hamsters were free of tumors, whereas therapy with [D-Trp6]LH-RH resulted in seven tumor-free animals, and combination of RC-160 and [D-Trp6]LH-RH resulted in eight tumor-free animals from groups of 15. Only preblastomatous lesions were found in these animals. Average tumor weight of animals in all peptide-treated groups, sacrificed 60 days after beginning the peptide treatment, was significantly lower than that of BOP controls. No significant differences were seen between the various peptide-treated groups. Histologically, analog-treated tumors of hamsters showed striking regressive changes characteristic of programmed cell death (apoptosis). This apoptosis presumably resulted from hormonal effects on tumor cells from prolonged treatment with these analogs of hypothalamic hormones. Our present data confirm the beneficial effect of long-acting microcapsules of [D-Trp6]LH-RH and RC-160 on pancreatic carcinoma and suggest a mode of action for these peptides. The feasibility of applying this treatment with analogs of hypothalamic hormones to human pancreatic carcinoma can be envisioned from these studies. PMID- 2564205 TI - Noise in neural networks: thresholds, hysteresis, and neuromodulation of signal to-noise. AB - We study a neural-network model including Gaussian noise, higher-order neuronal interactions, and neuromodulation. For a first-order network, there is a threshold in the noise level (phase transition) above which the network displays only disorganized behavior and critical slowing down near the noise threshold. The network can tolerate more noise if it has higher-order feedback interactions, which also lead to hysteresis and multistability in the network dynamics. The signal-to-noise ratio can be adjusted in a biological neural network by neuromodulators such as norepinephrine. Comparisons are made to experimental results and further investigations are suggested to test the effects of hysteresis and neuromodulation in pattern recognition and learning. We propose that norepinephrine may "quench" the neural patterns of activity to enhance the ability to learn details. PMID- 2564206 TI - The utilization of nitrogen for growth in mice fed blends of purified proteins. AB - Reasons for differences in growth of weanling mice fed different blends of purified proteins in diets apparently adequate in indispensable amino acids were investigated. Dietary nitrogen provided at increasing levels from a mixture of casein, gelatin, and zein, or from these plus gliadin and lactalbumin resulted in similar weight gains and efficiency of feed utilization in weanling mice. These responses were always lower (P less than 0.05) than for mice fed the control diet. When diets were based on true digestible nitrogen and amino acids, weight gains and feed efficiency decreased (P less than 0.05) as the dietary nitrogen level increased. High plasma glycine concentrations and high phenylalanine to tyrosine ratios were associated with high dietary glycine levels provided by gelatin. When a mixture of constant proportions of protein sources, predominantly casein, supplied most of the dietary nitrogen, small increases in weight gains and feed efficiency were observed as the nitrogen level in the diet increased. Varying the proportions of sources, which increased the percentage of gelatin, resulted in decreased gains in response to more dietary nitrogen. Replacing the amino acids found in gelatin by crystalline L-amino acids and replacing glycine by glutamic acid improved gains and feed efficiency (P less than 0.05) compared with gelatin-containing diets. These studies demonstrated that glycine at dietary concentrations of approximately 1-2%, either from gelatin or as the free amino acid, inhibited growth and feed utilization of growing mice. PMID- 2564207 TI - Novel autonomic neurotransmitters and intestinal function. AB - A wide variety of substances, including amines and peptides, have been detected within the complex neuronal pathways of the enteric nervous system using immunohistochemical techniques. In this article we have discussed some of the more recent data on the effects of these substances on intestinal activity. We have also commented on the many difficulties associated with ascribing neurotransmitter status to individual compounds. The technique of immunoblockade of neurogenic functional responses has been used in an attempt to identify some of the putative neurotransmitter substances. The search for selective antagonists continues. PMID- 2564208 TI - Effects of a specific beta 2-receptor blocker in neuroleptic-induced akathisia. AB - To assess the role of blockade of beta-receptor subpopulations in the treatment of neuroleptic-induced akathisia (NIA), the specific beta 2-antagonist ICI 118,551 was compared to placebo in a double-blind study. After a baseline evaluation on placebo, patients were treated with ICI 118,551 or placebo. Five of six patients treated with ICI 118,551 showed improvements in NIA, while only one of four patients improved on placebo. Patients were then treated openly with propranolol, a mixed beta 1, beta 2-antagonist. Compared to ICI 118,551, no further improvement on objective measures of akathisia was seen on propranolol. Mean subjective assessments of NIA declined on propranolol, but changes were variable and not statistically significant. PMID- 2564209 TI - Gastrin release in obese Zucker rats. AB - In this study, gastrin release in the obese Zucker rat was investigated by in vivo and in vitro experiments. Obese rats exhibited normal plasma gastrin levels at 3 weeks (preobese), were moderately hypergastrinemic at 3 months and severely hypergastrinemic at 5 months, compared to lean littermates. Following oral peptone, plasma gastrin levels doubled in both lean and obese rats. Basal and vagally stimulated gastrin release from perfused stomachs was greater in obese compared to lean rats and atropine had no effect on basal gastrin release in either group. Basal somatostatin release from the perfused stomach was found not to differ in both groups of animals. Morphological studies revealed an increase in the number of gastrin-containing G-cells in adult obese rats compared to lean littermates, but not in 3-week-old pups compared to lean littermates, indicating a strong correlation between cell number and plasma gastrin levels. These data indicate that the obese Zucker rat exhibits fasting hypergastrinemia in vivo, a condition which appears after weaning and increases in severity with age. Gastrin hypersecretion persists from the vascularly perfused stomach preparation. The basal hypergastrinemia of the obese Zucker rat is independent of a hyperactive postganglionic cholinergic drive but is associated with and probably causally related to an increase in antral G-cell numbers. PMID- 2564211 TI - Restriction fragment length patterns of DNA from parasitic nematodes of sheep. AB - High molecular weight DNA obtained from sheep parasitic nematodes Haemonchus contortus, Ostertagia circumcincta and Trichostrongylus colubriformis, was digested with various restriction endonucleases. Digestion with Eco R1 produced the most informative pattern of repeat sequence bands. H contortus adult or larval DNA produced bands of 2.7, 3.0 and 1.4 kb. O circumcincta adult or larval DNA had common 2.7 and 1.4 kb bands with adult specific bands of 2.2 and 0.9 kb and a larval specific 2.08 kb band. T colubriformis adults or larval DNA produced 2.7, 1.4 and 0.79 kb bands. These preliminary results show that restriction patterns of repeat sequence DNA may be useful for the identification of various trichostrongylid species parasitic for sheep. PMID- 2564210 TI - Effect of gamma-aminobutyric acid on bombesin-evoked release of somatostatin and gastrin from isolated rat stomach. AB - The effect of gamma-aminobutyric acid (GABA) on basal and bombesin (BBS) stimulated release of somatostatin (SLI) and gastrin from isolated perfused rat stomach was examined. In the control study, BBS at a dose of 10 nM significantly stimulated release of SLI and gastrin. Infusion of GABA (1-1000 nM) caused a depression of SLI release induced by BBS (10 nM) in a dose-dependent fashion. However, at doses used in this study GABA had no effect on either basal level of SLI and gastrin or BBS-elicited gastrin release. These results indicate that GABA can specifically modulate BBS-induced SLI release from rat stomach. PMID- 2564212 TI - [Receptor blockage: Nobel Prize for medicine 1988]. PMID- 2564213 TI - [Individualization of the antihypertensive treatment]. PMID- 2564214 TI - Molecular analysis of the human serum amyloid A (SAA) gene family. AB - We have assigned the human serum amyloid A (SAA) gene family to a 90 kb region on the short arm of human chromosome 11 (11p) by hybridization of defined genomic fragments of human SAA genes to DNA from rodent-human somatic cell hybrids and to large DNA fragments separated by transverse alternating field gel electrophoresis. We have also characterized SAA probe hybridization patterns in human DNA cleaved with restriction endonucleases Hind III, Pst I, BglII, TaqI, and XbaI and found invariant patterns except for a two-allele restriction fragment length polymorphism (RFLP) with Hind III. These studies show that the SAA gene family comprises at least three members in the haploid human genome and will be useful in identifying variant patterns and establishing linkage between members of the SAA gene family and other markers on chromosome 11. PMID- 2564215 TI - Dipeptidyl peptidase IV in human T lymphocytes. Impaired induction of interleukin 2 and gamma interferon due to specific inhibition of dipeptidyl peptidase IV. AB - Specific inhibitors of the membrane-bound dipeptidyl peptidase IV (DP IV) and polyclonal antibodies against this enzyme were used to investigate the relationships between DP IV activity and the production and action of T cell derived lymphokines. Production of interleukin 2 (IL-2) and gamma interferon by mitogen plus phorbol ester-stimulated mononuclear cells from human blood was found to be reduced in the presence of N-Ala-Pro-O-(nitrobenzoyl-)-hydroxylamine, epsilon-(4'-nitro) benzoxycarbonyl-Lys-Pro, and anti-(DP IV) immunoglobulin in a dose-dependent manner. Moreover, the proliferative response of mitogen-stimulated mononuclear cells to IL-2 is impaired in the presence of DP IV inhibitors. Therefore it is suggested that the membrane peptidase DP IV is involved in the induction and activation of cytokines controlling lymphocyte proliferation. PMID- 2564216 TI - Heme-dependent activation of soluble guanylate cyclase by nitric oxide: regulation of enzyme activity by porphyrins and metalloporphyrins. PMID- 2564217 TI - Prevalence of tardive dyskinesia in private psychiatric inpatients. AB - The authors in a one-week period examined inpatients at a large private psychiatric hospital to determine the prevalence of tardive dyskinesia. This population's characteristics are described and defined as different from those studied previously. Five of 90 patients (5.5%) who received neuroleptic drugs were judged to have tardive dyskinesia. The authors review the literature and discuss their findings. PMID- 2564218 TI - [Ultrasonic diagnosis of undescended testes. Methods and possibilities]. AB - 83 boys with 123 empty hemiscroti were assessed by clinical examination and ultrasound. After repeated clinical examination with the patient in general anesthesia 60 undescended testes were operated. We present the technical aspects of examination of undescended testes by ultrasound. When testicular position as established by ultrasound was compared with intraoperative finding position, ultrasound had a very high sensitivity, specificity and accuracy for testes positioned distal to the internal ring. The anatomical position of the testis at sonography and the patient's age provide objective evidence to discern retractile and truly undescended testes. PMID- 2564219 TI - Antagonism of bromobenzene-induced hepatotoxicity by the alpha-adrenoreceptor blocking agents phentolamine and idazoxan: role of hypothermia. AB - A recent study from our laboratory revealed that cotreating mice with the alpha adrenoreceptor antagonists phentolamine and idazoxan markedly diminished bromobenzene-induced hepatotoxicity. Subsequent studies also revealed that such cotreatment does not alter the pharmacokinetic disposition of bromobenzene in mice nor its bioactivation to reactive metabolites. In the present study, the possible role of hypothermia in the phentolamine antagonism of bromobenzene induced hepatotoxicity was investigated. Bromobenzene alone caused a significant, dose-related hypothermia. The high dosage regimen (10 mg/kg per dose) of phentolamine or idazoxan that had been found to be hepatoprotective in earlier studies potentiated this hypothermia and more than doubled the net decrease in core body temperature experienced by the animals. Placing mice receiving bromobenzene in an environment with an ambient temperature of 10 degrees C likewise increased the hypothermia experienced by animals receiving bromobenzene. The magnitude of the net change in core body temperature elicited by exposure to cold was similar to but slightly less than the net change produced by cotreatment with either alpha-adrenoreceptor antagonist and the magnitude of the hepatoprotection this procedure provided against bromobenzene hepatotoxicity was equivalent to that observed with phentolamine cotreatment. In contrast, a lower dosage regimen of either adrenoreceptor antagonist (2.5 mg/kg per dose) resulted in no additional hypothermia yet still produced a near maximal antagonism of bromobenzene-induced hepatotoxicity. Further, increasing the ambient temperature to 30 degrees C completely reversed the phentolamine-induced (10 mg/kg per dose) increase in hypothermia, but did not affect phentolamine's antagonism of the bromobenzene-induced changes in hepatic glutathione levels, serum alanine aminotransferase activity, or 24-hr mortality. Therefore, we conclude that while the hepatoprotective intervention of phentolamine can be mimicked by an exposure to cold that results in hypothermia, it is clear that alpha-adrenergic antagonists diminish the hepatotoxicity induced by bromobenzene by a mechanism that is independent of hypothermia. PMID- 2564220 TI - Failure of a CD18/anti-LFA1 monoclonal antibody infusion to prevent graft rejection in leukemic patients receiving T-depleted allogeneic bone marrow transplantation. AB - CD18 antibodies react with the common beta chain of the human leukocyte function antigen (LFA1)* and thus block the functions mediated by the three identified molecules in humans. A murine CD18 monoclonal antibody was infused in 8 leukemic patients receiving allogeneic T-depleted bone marrow transplantation in order to prevent graft rejection. This was part of the conditioning, including total-body irradiation and high-dose chemotherapy, given to all patients. To prevent graft versus-host disease the donor bone marrow T cells were depleted using complement mediated cytolysis or a ricin A conjugate immunotoxin, and cyclosporine or methotrexate were given posttransplant. A persistent level of free circulating anti-LFA1 antibody was detected in 5/8 patients. Despite this, 5 graft failures occurred, with 2 patients experiencing late rejection (days 60 and 97) following HLA-identical transplantation and 3 patients having no engraftment following haplo-mismatched transplant. One other patient died of early sepsis. Only 2 patients (who differed at 1 HLA locus from their donor) are alive with long-term complete chimerism (300 and 315 days). Transient inhibition of recipients' leukocyte functions with an anti-LFA1 antibody did not appear to facilitate engraftment of allogeneic T-depleted marrow transplantation for leukemias. PMID- 2564221 TI - [The fetal hydantoin syndrome]. AB - A boy with dysplasia/hypoplasia of nails and fingertips is reported. He was born to a woman who had been medicated during pregnancy with hydantoin for epilepsy. The clinical and radiological features of the fetal hydantoin syndrome are described. It is suggested that hydantoinexposed fetuses should be examined by ultrasound and serum folate monitored during pregnancy. PMID- 2564222 TI - Avian retroviral vectors derived from avian defective leukemia virus: role of the translational context of the inserted gene on efficiency of the vectors. AB - We have constructed retroviral vectors derived from the genome of avian erythroblastosis virus ES4 (AEV ES4). The neo selectable gene was substituted for the original v-erbA or v-erbB oncogenes of AEV, either in the same or in a different reading frames. Recombinant retrovirus were rescued and used to infect chicken embryo fibroblasts or quail QT6 cells. When the neo gene was inserted in the same reading frame as the original oncogene, we obtained (1) a high level of expression of the neo gene, (2) a balanced ration of both genomic and subgenomic RNAs, and (3) high titer recombinant viruses. Conversely, when the neo gene was inserted in a reading frame different from that of the original oncogene, we observed (1) a very low level of expression of the neo protein, (2) a predominance of the viral transcript used as translational template for the neo protein synthesis, and (3) low titer recombinant viruses. One of the vectors was used to transfer a human delta-globin gene into avian cells in culture without detectable rearrangement of this gene, but exhibited a deletion within the conserved noncoding region located between the two original oncogenes. Our data provide information for further construction of double expression vectors. Furthermore, three of the vectors would provide helpful tools to identify genetic elements of the virus genome involved in splicing regulation. PMID- 2564223 TI - WHO expert committee on drug dependence. Twenty-fifth Report. PMID- 2564224 TI - [Enzyme chemical studies of pericardial fluid]. AB - The authors examined the enzyme activities of LDH, CK, GOT, GPT and gamma-GT in blood-free pericardial fluid. The cases were divided into seven groups: a shot in the head with instant death, sudden cardiac death, poisoning, brain death with a long survival time, sudden infant death syndrome, and asphyxia. Taking all five enzymes into consideration, the cases of cardiac death differed significantly on the 1% level from the head shooting. However, concerning CK sudden cardiac death differed on the 5% level from the deaths as a result of poisoning. The wide range of the results, however, does not permit any reliable association of one single value with any of the respective groups. PMID- 2564225 TI - [Psychosomatic aspects of neuroleptic side effects]. AB - Using four clinical examples it is demonstrated that the neuroleptic induced side effects occasionally occur due to an interplay of pharmacological and psychological factors. On the one hand, psychic conflicts can in a more or less specific way foster the occurrence of certain side effects; on the other hand, the presence of side effects can enable the patients' psychic problems to become manifest. The clinician should always be on the alert regarding the possible psychosomatic nature of neuroleptic side effects and intervene psychotherapeutically if appropriate. PMID- 2564226 TI - The lupus anticoagulant in systemic lupus erythematosus. AB - The lupus anticoagulant was found in six of 41 unselected patients with systemic lupus erythematosus (14%). Three of these six patients had episodes of thrombosis. Thrombosis occurred in only one patient in the remainder of the series without the lupus anticoagulant. The lupus anticoagulant should be considered as one of the criteria for the diagnosis of systemic lupus erythematosus, and it may be a useful marker for those patients at risk from thromboembolism. It should be looked for in young adults with thrombotic episodes. PMID- 2564227 TI - Antibodies to the Borrelia burgdorferi flagellum in patients with scleroderma, granuloma annulare and porphyria cutanea tarda. AB - It is generally accepted that cutaneous Lyme borreliosis comprises erythema chronicum migrans, lymphadenosis benigna cutis, and acrodermatitis chronica atrophicans. In recent years the tick-borne spirochete Borrelia burgdorferi has been associated with a number of other cutaneous disorders. We therefore investigated sera from 175 patients with localized scleroderma (morphea) (n = 64), systemic sclerosis (n = 74), granuloma annulare (n = 16) and porphyria cutanea tarda (n = 21) with the new, highly sensitive and specific Borrelia burgdorferi flagellum ELISA assay. As controls (n = 297) served normal healthy volunteers and patients with other skin diseases. It was found that the distribution of individual antibody values and the median antibody levels were identical in controls and in patients with scleroderma, granuloma annulare and porphyria cutanea tarda. These data do not support the hypothesis of an etiological association between Borrelia burgdorferi infection and scleroderma, granuloma annulare or porphyria cutanea tarda. PMID- 2564228 TI - Tyrosinase activity in serum from patients with malignant melanoma. AB - A preparation procedure is presented for the determination of tyrosinase catalysed stereo-specific dopa oxidase activity in serum. Purification is obtained by separation on a Phenyl-Sepharose hydrophobic interaction column, followed by Con-A-Sepharose chromatography. Five out of seven sera from patients with widespread melanoma metastases were found to contain detectable quantities of tyrosinase. There was no tyrosinase activity in seven sera from patients with other malignancies, nor in six other control sera from individuals without malignancies. One serum which showed high tyrosinase activity was processed as above and studied by SDS-PAGE. A dopa-reactive band with an apparent MW of 66 kD was present in the gel, i.e. at the same place as that of the soluble tyrosinase of cultured human malignant melanoma cells. The protein was found to have the same pI at isoelectric focusing, and eluted in the same way from the preparation columns used, as did soluble tyrosinase. PMID- 2564229 TI - Isolation of human tyrosinase from cultured melanoma cells. AB - Tyrosinase was isolated from cultured melanoma cells using a procedure involving solubilization of the enzyme by means of Triton X-100, followed by different types of chromatography and tryptic digestion to make the enzyme soluble even in the absence of detergent. Starting with a membranous material containing 72 mg protein, 0.21 mg tyrosinase was obtained. The recovery of tyrosinase was 36% of the quantity found in the membranous starting material. In order to acquire a completely purified enzyme preparation suitable for amino acid sequence analysis, SDS-PAGE followed by blotting onto a polyvinylidene difluoride membrane was performed as a final step. The apparent molecular weight was found to be 66,000. Determination of the amino acids of the aminoterminal portion by automated Edman degradation showed the following sequence: His-Phe-Pro-Arg-Ala-X-Val-Ser-Ser-Lys Asn-Leu-Met-Glu-Lys-Glu-X-X-Pro-Pr o-The enzyme purified has an amino acid sequence identical with that of human tyrosinase deduced from c-DNA by Kwon et al. Striking similarities between our amino acid sequence and that predicted by Yamamoto et al. from mouse tyrosinase c-DNA were also observed. PMID- 2564230 TI - Serum selenium levels in patients with malignant melanoma. AB - The incidence of malignant melanoma of the skin has increased rapidly among white people during the last decade. Although the pathogenesis of malignant melanoma is not clear, epidemiologic data and experimental work indicates that ultraviolet (UV) radiation plays a critical role in tumorigenesis. Recent data implicate peroxidative reactions in UV-carcinogenesis and clearly demonstrate that selenium (Se) confers protection, in part by inducing cellular-free radical scavenging systems and by enhancing peroxide breakdown, thus enhancing the capacity of the cell to cope with oxidant stress. With this in mind, we investigated the Se status of 101 patients with malignant melanoma. Our study revealed significantly lower Se levels in the sera of melanoma patients than of controls. Although patients in all clinical stages had lower Se levels than the controls, patients in stage III (disseminated melanoma) had the lowest levels. Separate analysis of histogenetic subtypes of melanoma revealed that lentigo maligna melanoma (LMM) and superficial spreading melanoma (SSM) had the strongest association with depressed Se serum levels. Our results, showing for the first time that malignant melanoma is associated with low Se concentrations, are consistent with results of epidemiologic studies showing an inverse correlation between serum Se levels and certain cancers. PMID- 2564231 TI - Modulation of glutathione level in cultured human melanoma cells. AB - The effects of buthionine sulphoximine (BSO) treatment on cellular glutathione (GSH) content and on the cytotoxic action of menadione were investigated in cultured IGRI human melanoma cells. Addition of BSO (10(-8)-0.5 X 10(-3) M) to the cultures resulted in a dose- and time-dependent depletion of cellular GSH. BSO (10(-5) and 10(-6) M) did not influence cell multiplication up to 48 h, as determined by trypan blue staining. Menadione (3 X 10(-5) M) treatment decreased the cellular GSH concentration and also reduced cell number after a 24 h exposure. Its cytotoxicity was increased by BSO (10(-5), 10(-6) M), though the potentiating effect was moderate. PMID- 2564232 TI - Non melanoma skin cancer of the scalp. On the etiology. AB - In order to evaluate the relative significance of previous grenz-ray treatment for human non melanoma skin carcinogenesis, the files were studied of all patients treated for non melanoma skin cancer of the scalp (n = 82, male/female ratio 1.1) at the Department of Dermatology, the Finsen Institute, from 1976 to 1985. Fourteen patients, with a male/female ratio of 3.7, were treated for squamous cell cancer (SCC). Sixty-five patients, with a male/female ratio of 0.9, were treated for basal cell cancer (BCC). Twelve patients (15%, 11 with BCCs, 1 SCC), of which eight with psoriasis, were previously treated with grenz rays on the scalp, and two of them had not been exposed to additional skin carcinogens. Comparably, malignant conversion in sebaceous and verrucous nevi accounted for 9 cases or 11%. Characteristically, scalp cancers associated with previous grenz ray treatment were BCCs, the male/female ratio were less than 0.1 and two-thirds occurred in patients with multiple skin cancer. That grenz-ray related scalp cancers more often develop in females than in males was further confirmed by comparison to the sex distribution among patients treated on the scalp with grenz rays in the years 1950, 1960 and 1970 (p less than 0.01). (Accepted August 10, 1988.) PMID- 2564233 TI - Efficacy of topical treatment in psoriasis with MC903, a new vitamin D analogue. AB - In 10 in-patients with chronic plaque psoriasis, the antipsoriatic effect of MC903, a new synthetic analogue of vitamin D was evaluated. In each patient two symmetrical located psoriatic plaques were selected for the study. Topical treatment with MC903 cream (containing 1.2 mg MC903 per g cream) was compared with placebo cream in a double-blind, controlled, left-right, randomized way during 6 weeks of therapy. Compared with baseline, the clinical (erythema, scaling and infiltration) improvement was significant after 1 week of therapy with MC903 cream, while lateral comparison showed MC903 cream significantly better than cream base after 4 weeks of therapy (p less than 0.05). Measurements of skin blood flow by the laser Doppler technique in evaluating the disease activity was not superior to the clinical assessments. In 3 patients the psoriatic lesions treated with MC903 cream cleared completely during 6 weeks of therapy. No essential adverse reactions were observed. MC903 has a potent effect on cell proliferation and cell differentiation, but has minimal effect on calcium metabolism. It is concluded that this synthetic vitamin D analogue is potentially useful in the treatment of psoriasis. PMID- 2564234 TI - Prevalence of cement eczema in Denmark before and since addition of ferrous sulfate to Danish cement. AB - This is a study of the prevalences of chromate allergy and hand eczema among workers engaged in the manufacture of pre-fabricated concrete building components in Denmark in 1981 and again in 1987. In September 1981 the chromate content of cement manufactured and sold in Denmark was reduced to not more than 2 ppm (parts per million) of water-soluble chromate. This was accomplished by adding ferrous sulfate, thus increasing the cost of the cement by about 1%. There was a statistically significant decrease in the prevalence of chromate allergy and hand eczema following the addition of ferrous sulfate, but there was no change in the frequency of skin irritation. The economic benefit of adding ferrous sulfate was demonstrated by a decrease in the need for dermatological services and topical steroid treatment. Cement eczema as a result of chromate allergy is a common occupational dermatitis among workers in the building and construction industries and a reduction in the chromate content of cement would appear to be a reasonable preventive measure in areas where there is a large concentration of construction industries. PMID- 2564235 TI - Diffuse palmoplantar keratoderma associated with acrocyanosis. A family study. AB - Four members of a family, in which 8 suffered from diffuse palmoplantar keratoderma associated with an uncommon form of acrocyanosis, are reported. Acrocyanosis and palmoplantar keratoderma do not always occur together and, therefore, an autosomal dominant inheritance for this association is suggested. PMID- 2564236 TI - Atopic dermatitis and the indoor climate. The effect from preventive measures. AB - Nine patients with atopic dermatitis (AD) were clinically evaluated before and after moving to new houses with improved air exchange, low relative humidity and optimal temperature control. During a 2-year period three clinical and subjective assessments were performed each month of disease activity, and compared with changes in suspended and respirable dust particles, room temperature, air exchange rate, concentration of house-dust mites in bedrooms, and the concentration of organic solvents in the indoor air. Ten matched patients with AD, who did not move, served as a control group. The skin condition of patients moved improved significantly after moving. The indoor climate was improved on: 1) air exchange rate, 2) relative humidity, and 3) room temperature, but the amounts of house dust mites, respirable air particles and organic solvents were unchanged. The clinical and subjective improvement in AD could not be correlated to any single indoor environmental factor. The present investigation supports the current concept, that AD may be a multifactorial disease, and that the indoor climate may be a contributing factor affecting the eczema. PMID- 2564237 TI - Plasma cells in the dermal infiltrate of mycosis fungoides are of polyclonal origin. AB - There are several reports on the occurrence of immunoglobulin-producing B-cell lymphomas in Sezary's syndrome and mycosis fungoides, which may be the consequence of the helper activity of the neoplastic T-cells. Therefore we investigated skin biopsies of 50 patients with mycosis fungoides regarding the presence of plasma cells and their immunoglobulin profile. Nine of these patients had plasma cell nests, most frequently located at the lower edge of the infiltrate. IgE was detectable consistently, and IgG, IgM and IgA could also be demonstrated in the majority of these cases; kappa- and lambda-chains were present in equal amounts. Our results demonstrate polyclonal activation of plasma cells in a subgroup of mycosis fungoides patients. PMID- 2564238 TI - Efficacy and transdermal absorption of permethrin in scabies patients. AB - The clinical efficacy and transdermal absorption of permethrin, a new synthetic insecticide was investigated in ten scabies patients. All patients were successfully treated with one application of a cream, containing 5% permethrin. Apart from mild postscabies dermatitis no side-effects were observed. The mean weight of cream used per patient was 25 g (range 21-32; mean content of permethrin 1250 mg). The degree of permethrin absorption was assessed indirectly by determination of conjugated and unconjugated cis- and trans-CVA (a metabolite of permethrin) excretion in urine using two dimensional gas chromatography mass spectrometry. It was found that during the first 48 hours the mean estimated absorption was 6 mg (range 3-11), which is approximately 0.5% of the total dose. PMID- 2564239 TI - Ringworm-like late syphilides. AB - A peculiar form of late, superficial, serpiginous, non-ulcerative syphilides, leaving no visible scars, resembling clinically the ringworm of the glabrous skin, showing the granulomatous histopathological structure and relatively abundant T. pallidum, predominantly spread around the blood vessels is described. PMID- 2564240 TI - Distribution of CD1a-positive cells in psoriatic skin during the evolution of the lesions. AB - Normal skin and psoriatic lesions from 35 patients were investigated immunohistochemically with regard to the CD1a+ cell population (Langerhans' cells and indeterminate cells) in the epidermis as well as in the dermal infiltrate. In the normal-appearing skin, we found the regularly typical pattern of CD1a+ dendritic cells in suprabasal position, but in lesional skin of chronic psoriasis the CD1a+ cells were scattered in the acanthotic epidermis. In initial lesions, CD1a+ cells represent up to 50-60% of the infiltrating cells of the dermal compartment, in several cases being preferentially localized in the upper part of the papillar dermis close up to the epidermal CD1a+ cells in basal position, whereas in chronic psoriasis they represent less than 10%. These results suggest that in psoriasis vulgaris, CD1a+ cells actively migrate between the epidermis and the dermal vessels. PMID- 2564241 TI - Androgens in hirsute women referred for electroepilation. AB - One hundred and five hirsute women were examined. Elevated levels of at least one androgen were found in 58 women, most often free testosterone, dehydroepiandrosterone sulfate and delta 4-androstenedione. No significant correlation was found between level of androgens and hirsutism score or frequency of menstrual disturbances other than amenorrhea. Fifty-three women were examined at least 6 months after electroepilation. A significant decrease in hirsutism score and frequency of self-treatment was found. Scarring was not observed. Electroepilation performed by an experienced person should be recommended as an excellent treatment for hirsutism. PMID- 2564242 TI - Immunohistochemical reactivity of monoclonal antibodies generated after immunization of mice with cells from a psoriatic lesion. AB - Monoclonal antibodies were produced by immunizing mice with dispersed epidermal cells from a psoriatic plaque. We obtained 128 growing hybridoma clones. The immunohistochemical reactivity of the antibodies produced by these hybridomas was primarily screened on sections of skin biopsies from psoriatic plaques. Supernatants with positive staining were then tested on other dermatoses and human organs. Six supernatants stained infiltrating dermal cells and the whole epidermis in psoriatic lesions, but showed no reaction with sections from normal skin, ichthyosis, or acutely inflamed skin. With these antibodies, positive staining was also seen in some areas of the normal-appearing skin of patients with increasingly active psoriasis as well as after treatment in recently clinically healed lesions. A different and more heterogenous staining of epidermis with no or fewer infiltrating cells was seen in longstanding inflammatory disorders such as chronic eczema, neurodermatitis, lichen planus, discoid lupus erythematosus and mycosis fungoides. Scattered staining with these antibodies was in addition seen on infiltrating macrophage-like cells present also in other organs than skin and in some of the organs also other types of cells were stained, such as neck mucosal cells of the stomach, parts of duodenal mucosal cells, certain secretory cells in parotid gland, lung bronchiole and Hassall's bodies on the thymus. The cross-reactivity of antibodies to psoriatic lesions with other organs is of interest, since psoriasis might involve various tissues. Our findings support the idea that there exist unique cell surface antigens in psoriatic skin which might help elicit an immune reaction and/or be targets for such local immune reactions. PMID- 2564243 TI - Benzodiazepine antagonism by aminophylline. AB - A double-blind, randomised study was performed to investigate whether aminophylline could reverse the sedative effect of benzodiazepine and if it could shorten the observation time necessary after benzodiazepine sedation. Forty patients undergoing minor abdominal, urogenital or lower extremity surgery were given benzodiazepine to maintain a state of deep sedation after spinal or epidural analgesia was achieved. Postoperatively, the patients received either aminophylline, 110 mg, or physiological saline intravenously. The aminophylline treated patients showed a significantly more rapid reversal of sedation, but after 30 min there was no difference between the two groups. It is concluded that aminophylline antagonizes the sedative effect of benzodiazepine, but in routine benzodiazepine sedation, aminophylline will not shorten the necessary observation period after sedation. PMID- 2564244 TI - Effect of increased alpha-adrenergic activity on the blood pressure/cardiac output relationship in dogs. AB - The relationship between mean aortic blood pressure (MAP) and cardiac output (CO) was examined in anaesthesized, open-chest dogs during variations in pre-load with and without alpha-adrenergic stimulation with phenylephrine. When phenylephrine increased MAP to 200 mmHg, CO fell greatly and could not be increased by volume expansion. Left ventricular ultrasonic measurements and pressure recordings showed that the Frank-Starling mechanism was maximally activated. During vena cava obstruction CO and MAP fell proportionally. At a lower infusion rate of phenylephrine, MAP increased to 160 mmHg without a great reduction of CO. As in control experiments without phenylephrine infusion, CO could be increased by dextran/saline infusion and lowered about 20% below control by vena cava obstruction with no significant change in MAP; by further caval obstruction CO and MAP fell in proportion. Phenylephrine did not alter the relationship between aortic baroreceptor activity and MAP. The same MAP/CO relationships were obtained before and after bilateral vagotomy and nephrectomy. Caval obstruction and pacing tachycardia resulted in similar MAP/CO relationships despite different effects on left ventricular end-diastolic pressure. Thus, phenylephrine infusion may raise MAP to 200 mmHg but no cardiac reserve is left. During reduction of CO by caval obstruction, peripheral vascular resistance remains constant despite varying baroreceptor activity. At the lower infusion rate of phenylephrine, raising MAP to 160 mmHg, peripheral vascular resistance is constant at low CO, but at high CO the vasoconstrictive effect of phenylephrine is counteracted by a vasodilatory mechanism which seems to be flow-dependent. PMID- 2564245 TI - Priapism and neuroleptics: a case report. PMID- 2564246 TI - On-line small-bore chromatography for neurochemical analysis in the brain. PMID- 2564247 TI - Fabry disease: molecular genetics of the inherited nephropathy. AB - Originally described as a dermatologic curiosity by Fabry in 1898 and independently by Anderson in the same year, Fabry disease is now recognized as an inborn error of glycosphingolipid metabolism resulting from the defective activity of the lysosomal enzyme, alpha-galactosidase A (see Desnick and Sweeley for a comprehensive review). The enzymatic defect, transmitted by an X-linked recessive gene, leads to the accumulation of neutral glycosphingolipids with terminal alpha-galactosyl residues in the plasma and in the lysosomes of endothelial, perithelial, and smooth muscle cells of the cardiovascular-renal system and, to a lesser extent, in reticuloendothelial, myocardial, and connective tissue cells. Epithelial cells in the kidney, cornea, and other tissues contain the lysosomal depositions, as do the ganglia and perineural cells of the autonomic nervous system. The major accumulated substrate is globotriaosylceramide [galactosyl-(alpha 1----4)-galactosyl-(beta 1----4) glucosyl-(beta 1----1')-ceramide]; another substrate, galabiosylceramide [galactosyl-(alpha 1----4)-galactosyl-(beta 1----1')-ceramide] is deposited primarily in renal lysosomes. The clinical manifestations of Fabry disease are the sequelae of the anatomical and physiologic alterations produced by progressive glycosphingolipid deposition. Clinical onset of the disease in hemizygous males usually occurs during childhood or adolescence, with periodic crises of severe pain in the extremities (acroparesthesias), the appearance of the vascular cutaneous lesions (angiokeratoma), hypohidrosis, and the characteristic corneal dystrophy. With increasing age, the major morbid symptoms of the disease result from the progressive infiltration of glycosphingolipid in the cardiovascular-renal system. Death usually occurs from renal, cardiac, or cerebral complications of the vascular disease. Prior to the availability of treatment by renal transplantation or dialysis, the average age at death for affected males was about 40 years. Heterozygous females, who may exhibit the disease in an attenuated form, are most likely to have only corneal opacities. Previously, the diagnosis of affected hemizygous males and heterozygous females was based on clinical findings and the levels of alpha-galactosidase A activity in easily obtained sources, e.g., plasma and isolated lymphocytes or granulocytes. Because the gene encoding alpha-galactosidase A undergoes random X inactivation, the expressed level of enzymatic activity in females heterozygous for the disease gene may vary significantly, thereby making accurate carrier detection difficult.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2564248 TI - Genetic amyloidosis: recent advances. AB - The hereditary amyloidoses are usually associated with peripheral neuropathy and, for this reason, they have been called familial amyloidotic polyneuropathy (FAP). The neuropathy usually involves both the peripheral sensory motor and autonomic nervous systems, resulting in not only loss of sensation and motor function but also bowel dysfunction and orthostatic hypotension. In addition, most of the FAP syndromes involve other organ systems, in particular the renal and cardiac systems. Renal amyloidosis usually presents as nephrotic syndrome and later terminates in azotemia. Cardiac amyloidosis may present as congestive heart failure or arrhythmia. Whereas the neuropathy of hereditary amyloidosis may be the most prominent clinical feature, it is renal or cardiac disease that often dictates the length of survival. PMID- 2564249 TI - Molecular genetics and polycystic kidney diseases. AB - Polycystic kidney diseases (PKDs) comprise a group of common disorders, inherited as a monofactorial trait, usually dominant but sometimes recessive. The origin and mechanism of these diseases are unknown. Molecular genetics has now provided the means to seek the diseased gene(s), however, and, once found, to unravel the protein sequence and ultimately to understand the pathophysiology of these disorders. PMID- 2564250 TI - Test may predict which patients with HIV infection will develop AIDS. PMID- 2564251 TI - Electrical pacemakers of canine proximal colon are functionally innervated by inhibitory motor neurons. AB - Pacemaker activity in the canine proximal colon occurs at the submucosal and myenteric borders of the circular layer [Am. J. Physiol. 252 (Cell Physiol. 21): C215-C224 and C290-C299, 1987]. The present study investigated the neural regulation of rhythmic electrical activity. Spontaneous inhibitory junction potentials (IJPs) were observed in intracellular recordings from circular muscle cells near the myenteric border. The amplitudes of these events decayed with distance through the circular layer. Stimulation at the myenteric plexus surface evoked IJPs that mimicked the spontaneous events. Stimulation at the submucosal surface evoked IJPs in adjacent cells that were of shorter duration and of different waveform than myenteric IJPs. Amplitudes of IJPs evoked by stimulation near either surface decayed with distance from the site of stimulation. The decay functions for IJPs were essentially identical to the decay of spontaneous slow waves or myenteric potential oscillations. Spontaneous and evoked IJPs affected the amplitudes, durations, and patterns of ongoing rhythmic electrical activity. The data suggest that myenteric and submucosal pacemaker populations may be innervated by different populations of inhibitory nerve fibers. Innervation appears to be heterogeneous with dense populations of inhibitory nerve fibers predominantly located in the pacemaker regions. Neural regulations of pacemaker activity influences rhythmic electrical activity throughout the muscularis. PMID- 2564252 TI - Cytosolic free calcium in normal somatotropes: effects of forskolin and phorbol ester. AB - Digital imaging microscopy using the calcium-sensitive indicator probe fura-2 was combined with a reverse hemolytic plaque assay (RHPA) for growth hormone (GH) secretion. This technique allows dynamic measurements of the cytosolic free calcium concentration ([Ca2+]i) in individual pituitary somatotropes. Stimulation by growth hormone-releasing factor (GRF) increases, whereas somatostatin (SRIF) reduces [Ca2+]i in this cell type. [Ca2+]i increased in somatotropes when the cellular content of adenosine 3',5'-cyclic monophosphate (cAMP) was elevated by 1) activating cellular adenylate cyclase with forskolin (5 microM) and 2) treatment with the cAMP-analogues dibutyryl-cAMP (1 mM) or 8-bromo-cAMP (5 mM). The forskolin-induced calcium rise was abolished in the absence of extracellular calcium. This indicates that cAMP increases the influx of calcium into the cytosol and thereby stimulates hormone release. When forskolin was given in combination with SRIF (10 nM), [Ca2+]i decreased to the same level reached with SRIF treatment alone, indicating a site of action distal to the generation of cAMP. Activating protein kinase C with the phorbol ester 12,13-phorbol dibutyrate (PDB; 100 nM) increased [Ca2+]i as well. Again, this effect was dependent on extracellular calcium and blocked when PDB and SRIF were applied simultaneously. Combined stimulation with GRF plus PDB did not augment the response of [Ca2+]i over GRF treatment alone. PMID- 2564253 TI - Determinants of glutathione efflux and biliary GSH/GSSG ratio in perfused rat liver. AB - Utilizing the isolated perfused rat liver, we examined several factors influencing efflux of glutathione [reduced glutathione (GSH) and glutathione disulfide (GSSG)] into perfusate and bile, including the effects of perfusate composition, oxygen delivery to the liver, and acivicin (AT-125), an inhibitor of gamma-glutamyl transferase activity. When livers were perfused with a recirculating Krebs-Ringer bicarbonate buffer only 7-26% of released glutathione was excreted into bile, mainly in its oxidized form (71-90% as GSSG). In contrast, when 20% bovine red blood cells or 20% fluorocarbon emulsion were utilized as perfusates, biliary glutathione accounted for a larger fraction of total hepatic efflux (16-41%), and only 39-65% was excreted as GSSG. To determine whether O2 delivery to the liver could explain some of these differences, biliary and sinusoidal efflux of glutathione were measured as O2 delivery was varied by 1) increasing the perfusion flow rate, 2) altering the concentration of fluorocarbon emulsion (5, 10, and 20%), and 3) changing the PO2 (95% O2-5% CO2 vs. 50% O2-5% CO2-45% N2). Under all experimental conditions, an increase in O2 delivery was accompanied by an increase in bile flow and in the concentration and rate of glutathione efflux into bile but no significant change in sinusoidal efflux of glutathione. Hepatic tissue GSH and GSSG levels were not affected by the various treatments. When gamma-glutamyl transferase activity was inhibited with AT-125, biliary glutathione increased to levels of approximately 50% of total hepatic efflux in fluorocarbon-perfused livers, and only 24-29% of the glutathione was excreted as GSSG.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2564254 TI - Calcitonin gene-related peptide action on intestinal circular muscle. AB - Isolated segments of the guinea pig small intestine were used to examine the effect of calcitonin gene-related peptide (CGRP) on the motor activity of the circular muscle. CGRP (0.3-30 nM) initiated phasic contractions of the circular muscle due to stimulation of cholinergic neurons. Peristalsis, however, was inhibited by CGRP. A further analysis of this effect showed that CGRP had no inhibitory influence on the main, cholinergic, pathway of the ascending enteric reflex (AER) contraction, whereas the hexamethonium- and atropine-resistant pathways of the AER were blocked. The inhibition of the atropine-resistant AER resulted from an action of CGRP on nerves and, since it was antagonized by apamin, might be explained by a CGRP-induced activation of enteric inhibitory neurons. The direct relaxant action of CGRP on the longitudinal muscle was not affected by apamin. These findings indicate a heterogeneity and topical selectivity in the motor actions of CGRP on the gut and suggest that this peptide, when released from nerve endings within the intestine, plays a specific role in the regulation of intestinal motility. PMID- 2564255 TI - Neuropeptide Y augments adrenergic contractions at feline lower esophageal sphincter. AB - The purpose of this study was to determine the anatomic and physiological interactions between neuropeptide Y (NPY) and adrenergic transmitters at the feline lower esophageal sphincter (LES). Intraluminal pressures of the esophagus, LES, and gastric fundus were recorded in anesthetized cats. In a separate group of cats, gastroesophageal junctions were removed after locating the LES manometrically. Adjacent sections were stained with antibodies for NPY and tyrosine hydroxylase (TH). Indirect immunofluorescence revealed staining of TH- and NPY-like immunoreactive (LI) nerves in the circular muscle of the LES. Staining was also present for NPY-LI in longitudinal muscle, muscularis mucosa, periarterial nerves, and nerves and cell bodies of the myenteric plexus of the LES. In vivo, NPY produced a biphasic effect at the LES with an initial contraction (lasting approximately 30 s) followed by a relaxation (lasting approximately 3 min). NPY at the dose giving 50% of the maximum response (10(-6) g/kg) induced a contraction of 14.8 +/- 5.0 mmHg (P less than 0.05) followed by a decrease of basal LES pressure from 29.2 +/- 4.7 to 19.4 +/- 2.9 mmHg (P less than 0.01). Propranolol and phentolamine had no influence on either effect of NPY.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2564256 TI - Na-HCO3 cotransport and Na-H antiporter in chronic respiratory acidosis and alkalosis. AB - Renal acidification in renal proximal tubule is thought to be mediated by luminal Na-H antiporter and the HCO3- generated by this antiporter is removed from the cell by a basolateral Na-HCO3 cotransporter. To study the effect of respiratory acid-base disorders on these transport systems, we have measured the Na-HCO3 cotransport in basolateral membranes and Na-H antiporter in luminal membranes in control rabbits, rabbits exposed to 10% CO2 (chronic hypercapnia), and rabbits exposed to 10% O2-90% N2 (chronic hypocapnia). The Vmax of HCO3(-)-dependent 22Na uptake was significantly higher in chronic hypercapnia than controls (2.54 +/- 0.03 vs. 1.18 +/- 0.21 nmol.mg protein-1.3 s-1, P less than 0.001). Likewise, the Vmax of the Na-H antiporter was also increased compared with controls (924.9 +/- 42.1 vs. 549.1 +/- 62.8 fluorescence units (FU).300 micrograms protein-1.min-1). In chronic hypocapnia, the Vmax of Na-HCO3 cotransport was lower than controls (0.72 +/- 0.11 vs. 1.18 +/- 0.21 nmol.mg protein-1.3 s-1, P less than 0.05). There was no difference, however, in the Vmax of the Na-H antiporter between hypocapnia and control (524.2 +/- 24.3 vs. 549.1 +/- 62.8, FU.300 micrograms protein-1.min-1). The Vmaxs of the Na-HCO3 cotransport and of the Na-H antiporter in hypocapnic, control, and hypercapnic rabbits were linearly related (r = 0.81), suggesting a simultaneous adaptation of the two systems in respiratory acid-base disorders.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2564257 TI - Interstitial dynamics in rats with early stage experimental cirrhosis of the liver. AB - Pathogenesis of edema in cirrhosis of the liver is still incompletely understood. The present study was designed to examine interstitial fluid dynamics in cirrhotic, non-ascitic rats, measuring interstitial fluid pressure by means of a subcutaneous plastic capsule in basal conditions during extracellular fluid volume expansion with Ringer solution and during albumin infusion. Urine flow and sodium excretion and plasma and interstitial fluid volumes were simultaneously measured. Cirrhotic rats exhibited reduced urine flow and sodium excretion, both in basal conditions and in response to expansion maneuvers. Plasma and interstitial fluid volumes were higher in cirrhotic than in control animals. Remarkable alterations were present in capsular pressures in cirrhotic rats. In the control rats, basal capsular pressure values were negative, and they increased after Ringer infusion and markedly decreased with albumin infusion. In contrast, in cirrhotic rats, basal capsular pressures were in the positive range and they remained nearly constant during ringer infusion and albumin administration. These results suggest that in cirrhotic rats there are significant alterations in systemic interstitial dynamics, even before ascites formation. Altered systemic capillary dynamics may therefore be important early changes that precede and thus contribute to the formation of edema in cirrhosis. PMID- 2564258 TI - Neuronal histamine modulates feeding behavior through H1-receptor in rat hypothalamus. AB - To clarify the physiological role of hypothalamic neuronal histamine in control of food intake, ingestive behavior and neuronal activity were investigated under blockade or diurnal fluctuation of hypothalamic neuronal histamine. Histamine H1- but not H2-receptor antagonist potently induced feeding in a dose-related manner after infusion into rat third cerebroventricle at 1100 h. Elicitation was attenuated after infusion at 1940 h when histamine content in the hypothalamus was low and was abolished after intraperitoneal pretreatment with 0.11 mmol alpha fluoromethylhistidine (alpha-FMH), a specific suicide inhibitor of histidine decarboxylase. Electrophoretic application of a histamine H1-receptor antagonist to ventromedial hypothalamic neurons specifically suppressed activities of glucose-responding neurons that are related to food intake. The suppressive effect was also attenuated by intraperitoneal pretreatment with alpha-FMH. These results suggest that feeding induced by histamine H1-receptor antagonists is due to blockade of neuronal histamine at the site of histamine H1-receptors, at least in part, in the ventromedial hypothalamus and that diurnal fluctuation of feeding behavior may reflect circadian variation of neuronal histamine level. PMID- 2564259 TI - Effects of althesin and urethan-chloralose on neurohumoral cardiovascular regulation. AB - The cardiovascular effects of althesin (ALT) and urethan-chloralose (UC) anesthesia were compared in conscious, chronically instrumented rats. Althesin had no effect on arterial pressure or base-line resistance in the renal, superior mesenteric, and hindquarters vasculatures but increased heart rate. In contrast, UC decreased arterial pressure, heart rate, and mesenteric resistance. Although UC attenuated depressor responses to nitroglycerin, neither anesthetic significantly altered regional vascular reactivity to intravenous phenylephrine and nitroglycerin. The cardiac chronotropic baroreflex was examined by comparing the slope of the curves relating maximal changes (delta) in heart rate (pulse interval) that occurred at the point coinciding in time with the maximal changes in mean arterial pressure produced by phenylephrine and nitroglycerin. Neither anesthetic significantly altered the baroreflex slope (delta pulse interval/delta mean arterial pressure) for pressor and depressor stimuli. Both anesthetics attenuated the sympathoexcitatory response to cerebroventricular angiotensin II, although ALT had less of a depressive effect (pressor response during ALT and UC = 65 and 30%, respectively, of conscious). Plasma renin activity (PRA) and the hemodynamic response to peripheral angiotensin-receptor antagonism were significantly increased (PRA by almost 6-fold) during UC, whereas ALT was without effect. Similarly, UC but not ALT induced vasopressin-dependent vascular tone. Ganglionic blockade indicated that peripheral neurogenic tone was not altered by ALT anesthesia. These data suggest that althesin produces fewer hemodynamic disturbances than urethan-chloralose and largely maintains cardiovascular regulation intact. PMID- 2564260 TI - Graded neuropsychological impairment and elevated gamma-glutamyl transferase in chronic alcoholic men. AB - This study hypothesizes that distinct biochemical and metabolic disturbances associated with liver injury may be related to specific cognitive changes in alcoholics. In 132 alcoholic men admitted to an alcohol treatment program, increases in gamma-glutamyl transferase (GGT) values were correlated with impairment in several measures of visuoperceptual and visuoconceptual functioning. The association between plasma levels of GGT and neuropsychological performance was independent of the relative contribution of other laboratory measures of liver injury and of alcohol consumption histories. These observations support the hypothesis that elevated levels of GGT are distinctly associated with neuropsychological deficits and suggest that possible mechanisms beyond severe hepatic dysfunction and alcohol consumption underlie cognitive deficits in alcoholics. PMID- 2564261 TI - The cardiovascular effects of mivacurium chloride (BW B1090U) in patients receiving nitrous oxide-opiate-barbiturate anesthesia. AB - The dose-effect relationship of mivacurium chloride on arterial blood pressure, heart rate, and plasma histamine was determined in 97 consenting ASA physical status I-II patients receiving nitrous oxide-oxygen-opiate-barbiturate anesthesia. In the absence of surgical stimulation during steady state anesthetic conditions with controlled ventilation, average maximum change in tachograph counted heart rate was 7% or less after 10-15-s injection of mivacurium at all doses from 0.03 to 0.30 mg/kg. Average peak change in mean arterial pressure measured via radial arterial catheter was 7% or less after all doses from 0.03 to 0.15 mg/kg. Transient (0.2-4.5 min) decreases in arterial blood pressure were noted after 10-15-s injection in some patients at 0.20, 0.25, and 0.30 mg/kg. When they occurred, these changes were usually accompanied by facial erythema lasting 2-5 min and were correlated with increases in plasma histamine level (P less than 0.001). Facial erythema, decrease in blood pressure, and elevation of histamine level were all accentuated by increasing the dose of mivacurium and by more rapid injection of the drug. For example, mean blood pressure decreased an average of 13% after injection of mivacurium 0.25 mg/kg over 10-15 s. In contrast, during administration over 30 and 60 s of this dose, arterial pressure decreased 7.6 and 1.5%, respectively (P less than 0.001, 10-15 s vs. 60-s injection). Average peak histamine level, which increased to 132% of control after administration of 0.25 mg/kg over 10-15 s, did not change after injection over 60 s.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2564262 TI - Muscle pain occurs after outpatient laparoscopy despite the substitution of vecuronium for succinylcholine. AB - The occurrence, location, and severity of muscle pain were determined when vecuronium was used in lieu of succinylcholine during outpatient laparoscopy. Postoperative muscle pain, in 11 body parts, was assessed by a linear analogue scale questionnaire that was completed by each patient on the evening of surgery and for the next three mornings. All patients had general endotracheal anesthesia with nitrous oxide, thiopental, and fentanyl. Succinylcholine 1.5 mg/kg (3-4 min after 3 mg of d-tubocurarine) was given to 14 patients for tracheal intubation and then by infusion for additional muscle relaxation. Another 14 patients received vecuronium 50 micrograms/kg iv as the only muscle relaxant used; all of these patients had residual neuromuscular blockade antagonized with glycopyrrolate 7 micrograms/kg and edrophonium 0.5 mg/kg iv. Both groups were similar in age, weight, length of procedure, time to discharge, and amount of thiopental and fentanyl used (P greater than 0.05). No difference was noted in either group with respect to the severity of pain by body part over time. Mean total body pain scores were generated for each group at all four intervals as an alternate type of analysis. No statistical significance was demonstrated by a Student's t test in any group at any interval sampled. The authors failed to demonstrate that the substitution of vecuronium for succinylcholine lowers the incidence of myalgia when used in outpatient diagnostic laparoscopy. They refrain from concluding that vecuronium contributes to postanesthetic myalgia, but feel justified in stating that the avoidance of succinylcholine did not lower the severity or occurrence of muscle pains after laparoscopy when vecuronium was used in its place. PMID- 2564263 TI - A comparison of the neuromuscular blocking and autonomic effects of two new short acting muscle relaxants with those of succinylcholine in the anesthetized cat and pig. AB - The actions of two new steroidal nondepolarizing neuromuscular blocking agents, structurally related to vecuronium, have been compared with those of succinylcholine in anesthetized cats and pigs. Both new compounds (Org 7617 and Org 9616) exhibited properties typical of nondepolarizing relaxants in each species. Org 9616 was one-fifth (ED50 cat tibialis 154 micrograms.kg-1) and Org 7617 was one-tenth (ED50 cat tibialis 287 micrograms.kg-1) as potent as vecuronium as a neuromuscular blocking drug. Both compounds produced rapidly developing muscle relaxation in cats that, like that of succinylcholine, was transient in time course (onset/duration of action--tibialis anterior muscle: Org 7617 1.6/3.9 min; Org 9616 1.5/4.3 min; succinylcholine 1.7/5.7 min). In pigs that were used as a predictor of duration of action in humans, both Org 7617 and Org 9616 also produced short-lived neuromuscular blockade. The neuromuscular blocks produced by Org 7617 and Org 9616 were readily reversed by neostigmine. Both compounds blocked the heart rate responses to vagal stimulation at doses higher than those required to produce neuromuscular block. The vagal:neuromuscular blocking ratio for Org 7617 was 10, and for Org 9616 was 17. These compare to approximate published values for vecuronium, atracurium, and pancuronium of 60, 25, and 3, respectively. Ganglion block was only seen at 30-60 times the neuromuscular blocking doses. Both compounds produced a decrease in arterial blood pressure. This was more pronounced with Org 7617. In addition, Org 9616 produced a slight increase in heart rate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2564264 TI - Effects of nitrous oxide and halothane on mu and kappa opioid receptors in guinea pig brain. AB - The effects of two general anesthetics, nitrous oxide and halothane, and oxygen on mu and kappa opioid receptor subtypes from guinea-pig brain were investigated. mu receptor binding was defined using [3H]dihydromorphine as the ligand. Nitrous oxide (100%) and halothane (2%) decreased the [3H]dihydromorphine binding affinity (Kdair = 0.87 nM, KdN2O = 1.45 nM, Kdhalothane = 2.30 nM) without affecting the density of binding sites. A decrease in the [3H]dihydromorphine binding affinity without influence on the density of binding sites was also observed in the presence of 100% oxygen (KdO2 = 1.40 nM). kappa receptor binding was defined using [3H](-)ethylketocyclazocine as the ligand, in the presence of 100 nM D-ala2-D-leu5-enkephalin and 30 nM morphine. While 100% nitrous oxide caused a slight decrease in [3H](-)ethylketocyclazocine binding affinity (Kdair = 0.24 nM, KdN2O = 0.31 nM) and a substantial decrease in the density of binding sites (Bmaxair = 115 fmol/mg protein, BmaxN2O = 84 fmol/mg protein), halothane dramatically affected both the affinity (Kdhalothane = 0.70 nM) and density (Bmaxhalothane = 38 fmol/mg protein). Oxygen (100%) reduced [3H]dihydromorphine binding affinity. Differential effects of two anesthetics on the same receptor or distinct actions of the same anesthetic on different receptors could indicate the presence of specific targets for anesthetics at the membrane level. Conversely, effects of volatile anesthetics on opioid receptors could reflect a non-specific perturbation of the lipidic and proteinaceous components of the membranes. PMID- 2564265 TI - The impact of prednisone in life-threatening idiopathic anaphylaxis: reduction in acute episodes and medical costs. AB - Two patients with frequent episodes of well documented life-threatening idiopathic anaphylaxis were studied. Despite prior treatment with antihistamines and self-administered epinephrine, both patients continued to have episodes of anaphylaxis. At the time of presentation to the Northwestern University Allergy Service, prednisone and antihistamines with or without sympathomimetics were started. Described in this report are the substantial reductions in the frequency of hospitalizations, emergency room visits, and anaphylactic episodes after our medical management was instituted. PMID- 2564266 TI - Single-dose study of the effect of terfenadine on theophylline absorption and disposition. AB - Terfenadine, a nonsedating antihistamine, and theophylline, a methylxanthine used extensively in the treatment of asthma, have potential for frequent concomitant administration. A single-dose pharmacokinetic study was performed to investigate the potential for a drug-drug interaction affecting serum theophylline concentrations. No statistically significant effect on theophylline absorption or elimination was found when 60 mg of terfenadine was concomitantly administered with 4 mg/kg of theophylline in 17 normal healthy male volunteers. The results of this single-dose study suggest there is no significant acute pharmacokinetic interaction affecting serum theophylline concentrations when terfenadine and theophylline are administered concomitantly as single isolated doses. Further investigation is needed to determine if repeated administration of terfenadine is equally without effect on theophylline absorption or disposition. PMID- 2564267 TI - Effects of inhaled thiazinamium chloride on histamine-induced and exercise induced bronchoconstriction. AB - The protective efficacy of aerosolized thiazinamium chloride (TC) against histamine-induced and exercise-induced bronchoconstriction was evaluated in 15 subjects with stable, mild asthma. Following reproducible bronchoprovocation with these stimuli, each subject underwent randomized, double-blind, crossover pretreatment with single doses of nebulized TC (300, 600, and 900 micrograms), placebo, and an active control drug (metaproterenol or cromolyn), followed by histamine or exercise challenge (two separate protocols). The results indicated that all doses of TC significantly blocked histamine-induced bronchoconstriction as compared with placebo. Overall, aerosolized TC was ineffective in blocking exercise-induced bronchoconstriction, although 900 micrograms TC tended to be more effective than placebo. Thiazinamium (900 micrograms) produced a modest bronchodilator effect. No clinically significant adverse effects related to TC occurred. We conclude that aerosolized TC is effective in attenuating histamine induced but not exercise-induced bronchoconstriction in the doses studied. Further studies are warranted to evaluate the role of TC in asthma therapy. PMID- 2564268 TI - Fatal Portuguese man-o'-war (Physalia physalis) envenomation. AB - A fatal case of Physalia physalis (Portuguese man-o'-war) envenomation occurred on the Florida Atlantic coast in 1987. Despite appropriate beachside first aid, the patient was conscious only several minutes before having primary respiratory arrest and, later, cardiovascular collapse that resulted in death. Discharged nematocysts were still visible on the injured stratum corneum five days after envenomation. Additional treatment maneuvers suggested by this case include testing the tentacle fragments found on the victim's skin before their removal to ensure that nematocyst firing has been counteracted. We document the first human fatality caused by P physalis envenomation. PMID- 2564269 TI - Somatostatin and a long-acting analogue, octreotide acetate. Relevance to dermatology. PMID- 2564270 TI - Flavivirus entry into cultured mosquito cells and human peripheral blood monocytes. AB - The entry modes of Japanese encephalitis (JE) and dengue-2 (DEN-2) viruses into C6/36 mosquito cells and of DEN-2 virus into human peripheral blood monocytes in vitro were studied. Inoculation of either JE or DEN-2 virions into C6/36 cells resulted in direct penetration of the virions into the cytoplasm at the cell surface in 3 stages. At stage 1, virions attached to the plasma membrane of host cells by their envelope spikes; at stage 2, the virion envelopes approximated to and eventually overlapped the host plasma membrane, and in the process the plasma membrane at the attachment sites dissolved; and, at stage 3, virions penetrated into the cytoplasm through the plasma-membrane disruptions created at the adsorption sites. Virions themselves apparently disintegrated at or near the penetration sites, for no virions were seen in the deeper cytoplasm. Coated pits did not form at the virion attachment sites, and virion-containing vesicles were not found in the cytoplasm. In the entry of DEN-2 virus into human peripheral blood monocytes, virions were found, adsorbed onto the external surface of the plasma membrane and attached to the luminal surface of macropinocytic vacuolar membranes. The latter apparently occurred as the result of ruffling and macropinocytic activities of the cells. At both sites virions penetrated into the cytoplasm through the plasma or vacuolar membrane in the same manner as they did through the plasma membrane of C6/36 cells. No evidence of viral entry by receptor-mediated endocytosis was observed. Implications of the entry mode of the mosquito cell-generated DEN-2 virus into human peripheral blood monocytes to an early process of natural, mosquito-transmitted infection is discussed. PMID- 2564271 TI - Physical mapping of the genomic heterogeneity of isolates of equine herpesvirus 2 (equine cytomegalovirus). AB - The BamHI, EcoRI, and HindIII physical maps of the genomes of 14 isolates of equine herpesvirus 2 (EHV 2) were determined by Southern blot analysis using DNA fragments of a previously mapped EHV 2 strain 86/67. No two isolates had identical maps for all 3 enzymes, the number of differing cleavage sites between pairs of isolates varying from 3 to 21. Overall 75 cleavage sites were mapped, of which 40 were variable. Cleavage sites occurred throughout the genome, including within the terminal repeat regions. Additionally, fragment length polymorphisms, independent of cleavage site loss or gain, were mapped to 5 regions of the genome, 4 of which occurred within the terminal repeat regions. PMID- 2564272 TI - Excitotoxins: a possible new mechanism for the pathogenesis of ischemic retinal damage. PMID- 2564273 TI - Preliminary purification and characterization studies of a low molecular weight, high affinity cytosolic lead-binding protein in rat brain. AB - Carrier-free 203Pb has been used to label high affinity lead-binding proteins in rat brain cytosol to allow their initial characterization. The low molecular weight 203Pb-protein complex collected from a Sephadex G-75 column eluate has been further purified by Sephadex DEAE chromatography and then partially characterized. The protein has a molecular weight of 23,000 daltons as determined by SDS polyacrylamide gel electrophoresis and significant levels of glutamic acid (9.3%), aspartic acid (10.8%) and cysteine (9.4%). Western blot studies conducted using the polyclonal antibody to the renal lead-binding proteins showed a lack of reactivity, indicating that the brain protein is immunologically distinct from that found in the kidney. PMID- 2564274 TI - [Arg292----Val] or [Arg292----Leu] mutation enhances the reactivity of Escherichia coli aspartate aminotransferase with aromatic amino acids. AB - Arg292 of E. coli aspartate aminotransferase was substituted with valine or leucine by site-directed mutagenesis. In comparison with the wild-type enzyme, either of the mutant enzymes showed a decrease by over 5 orders of magnitude of kcat/km values for aspartate and glutamate. This supports the contention that Arg292 is important for determining the specificity of this enzyme for dicarboxylic substrates. In contrast, mutant enzymes displayed a 5- to 10-fold increase in kcat/Km values for aromatic amino acids as substrates. Thus, introduction of an uncharged, hydrophobic side chain into position 292 leads to a striking alteration in substrate specificity of this enzyme, thereby improving catalytic efficiency toward aromatic amino acids. PMID- 2564275 TI - Opossum kidney contains a functional receptor for the Escherichia coli heat stable enterotoxin. AB - The Escherichia coli heat-stable enterotoxin (ST1 or STa) binds to specific receptors on mammalian intestinal brush border membranes, and stimulates guanylate cyclase in those membranes. We have found a similar signal transduction system in brush border membranes prepared from kidney cortex of the American opossum (Didelphis virginiana, and in a cell line (OK cell) derived from that tissue. Activation of guanylate cyclase by ST1 is therefore not limited to intestinal cells. Furthermore, since it is unlikely that ST1 which is produced in the intestinal lumen, would have access to kidney receptors, this suggests the existence of an endogenous peptide resembling ST1, at least in marsupials. PMID- 2564276 TI - Behavior of mother rats in conflict tests sensitive to antianxiety agents. AB - Previous studies of freezing and open-field activity have demonstrated that lactating rats are less fearful or less anxious than nonpregnant ones. The purpose of this investigation was to observe the behavior of mother rats in conflict tests, which are frequently used in studies on the neurobiology of anxiety. In the punished drinking test, in which licking from a water spout is punished by electric shocks, mothers (observed on Day 1 postpartum following 24 hr of water deprivation) were found to drink more than virgins. Mothers (Day 1 postpartum) also consumed more food than controls in an unfamiliar open field. In contrast, no difference between mothers (Day 5 postpartum) and virgins was present in the exploration of an electrified shock probe. The largest maternal anticonflict effects in the drinking and feeding tests were recorded when the females were tested with their pups. Increased punished drinking was also observed in virgin rats treated with the anxiolytic benzodiazepine midazolam. Water-deprived virgins and mothers did not differ in the shock titration test, a result suggesting that diminished pain reactivity was unlikely to account for the increased punished drinking in mothers. Moreover, females in late pregnancy, which are hypoalgesic (Gintzler, 1980), did not lick more than virgins in the punished drinking test. Following 24 hr of water deprivation, unpunished drinking was higher in lactating females than in virgins, so the increased acceptance of punishment by mothers might have been due to their being more thirsty than virgins.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2564277 TI - Antagonism of NMDA receptors impairs acquisition but not retention of olfactory memory. AB - Prompted by evidence pointing to a key role of the N-methyl-D-aspartate (NMDA) receptor system in the induction of long-term potentiation and possibly in the formation of some types of memory, we examined the effect of chronic intraventricular administration of D-amino-phosphono-valeric acid (AP5), a competitive NMDA receptor antagonist, on olfactory discrimination and avoidance learning. These two tasks were selected because they are affected to very different degrees by damage to the hippocampus and other telencephalic structures rich in NMDA receptors. Twenty rats previously trained to solve a series of discriminations between two simultaneously presented odors were infused with either 20 mM D-AP5 or saline (n = 10 per group) for 14 days. An important and unusual feature of the paradigm was that it permitted a comparison of drug effects on acquisition of new discriminations versus retention of old ones. Animals treated with AP5 made significantly more errors than did saline controls in acquiring discriminations between low-intensity odors presented with long intertrial intervals (ITIs). However, no deficit was observed when short ITIs (less than 2 min) or strong odors were used. Animals treated with AP5 had no difficulty in recognizing odors on which they were trained before administration of the drug. After exhaustion of the pumps, performance of the AP5 group was indistinguishable from that of the control group. One-way active avoidance learning was not affected by chronic infusion of AP5. Several possibilities are discussed that could account for the selective olfactory learning deficit.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2564278 TI - Sympathetic nervous system and hypertension. Therapeutic perspectives. AB - The sympathetic nervous system has a key role in the physiological regulation of the circulation and may also mediate some of the factors that cause or sustain clinical hypertension. Most classes of antihypertensive drugs produce at least part of their effects by inhibitory actions on the sympathetic pathways. The centrally acting agents and the peripheral alpha-adrenergic blockers are the most specific currently used sympathetic inhibitors. The available alpha-blockers, prazosin and terazosin, effectively reduce blood pressure when used as monotherapy or in combination with other antihypertensive drugs. When given once daily, the actions of terazosin persist throughout the full 24-hour period, including the important midmorning hours when there may be heightened sympathetic activity. The incidence of symptomatic adverse effects with the alpha-blockers is low. Because they specifically block peripheral alpha-receptor-mediated vasoconstriction, the alpha-blockers decrease total peripheral resistance at rest and do not antagonize the appropriate vasodilatory responses to exercise. They maintain or increase blood flow in regional circulations; cerebral blood flow in elderly patients is unchanged despite significant decreases in systemic blood pressure. In contrast to other drug classes that can adversely affect the lipid profile, the alpha-blockers slightly decrease total cholesterol concentrations. Recent studies also have shown that terazosin and prazosin can promote decreases in echocardiographically measured left ventricular wall thickness and muscle mass in hypertensive patients with left ventricular hypertrophy. Thus, this type of sympathetic blockade not only produces antihypertensive effects, but additionally may have a beneficial impact upon other cardiovascular risk factors. PMID- 2564279 TI - Is vecuronium a long-acting neuromuscular blocking agent in neonates and infants? AB - Vecuronium was used as the only neuromuscular blocking agent in 81 paediatric patients (neonates to adolescents) during fentanyl and nitrous oxide anaesthesia. The thenar electromyogram was used to monitor neuromuscular blockade. Neonates and infants had a mean requirement of vecuronium 105 micrograms kg-1 during the first 1 h of anaesthesia, to establish and maintain 90-98% neuromuscular blockade, compared with a mean requirement of 217 micrograms kg-1 for children aged 3-10 yr (P less than 0.05). Vecuronium 100 and 150 micrograms kg-1 maintained neuromuscular blockade greater than 90% for 59 and 110 min, respectively, in neonates and infants, but only for 18 and 38 min in children and for 37 and 68 min in adolescents (P less than 0.05). Vecuronium may be regarded as a long-acting neuromuscular blocking agent in patients aged less than 1 yr. PMID- 2564280 TI - Haemodynamic effects of vecuronium. AB - Three hundred and twenty adults (ASA grade I, both sexes), received diazepam 10 mg by mouth (50% received atropine 7 micrograms kg-1 in addition) i.m. 45 min before operation. Patients were then allocated randomly to undergo general anaesthesia with either a nitrous oxide-neurolept technique or nitrous oxide halothane. Vecuronium was administered to 50% of the patients in each anaesthetic group and heart rate and arterial pressure were monitored. Vecuronium did not influence heart rate, or systolic or diastolic arterial pressures. PMID- 2564281 TI - Histiocytoma cutis: a tumour of dermal dendrocytes (dermal dendrocytoma). AB - The histogenesis of histiocytoma (dermatofibroma) was investigated using new antibodies that demonstrate factor XIIIa (FXIIIa) positive cells and the monocyte macrophage cell series (MAC 387), in formalin fixed tissue. The distribution of S100 protein, vimentin and Ulex europaeus agglutinin I (UEA-I) were also studied. The antibody against FXIIIa labelled the normal dermal population of fixed connective tissue cells (dermal dendrocytes) emphasizing their dendritic processes; cells that are widely distributed, but are most numerous in the papillary dermis. In contrast, the antibody, MAC 387 against monocyte derived macrophages, did not label this cell population. In 30 histiocytomas, intense labelling for FXIIIa was found peripherally, but labelling was rather weaker in cells situated centrally. Only a few cells labelled for S100 protein and with the monoclonal antibody MAC 387, but vimentin positivity was universal. UEA-I labelled only vessels in histiocytomas. However, FXIIIa labelling was negative in 16 dermatofibrosarcomas, three keloids and several other fibroproliferative lesions. These contrasting results with FXIIIa labelling have practical implications for diagnosis of benign and malignant spindle cell tumours of the skin. We conclude that histiocytomas (dermal dendrocytomas) take their origin from dermal fixed connective tissue cells, (dermal dendrocytes), but other spindle cell tumours may differ in histogenesis. PMID- 2564282 TI - Synthesis of pyrene derivatives of cerebroside sulfate and their use for determining arylsulfatase A activity. AB - Two fluorescent derivatives of cerebroside sulfate ('sulfatide') have been synthesized and used as substrates for determining arylsulfatase A activity. These were 12-(1-pyrene)dodecanoyl cerebroside sulfate (P12-sulfatide) and 12(1 pyrenesulfonylamido)dodecanoyl cerebroside sulfate (PSA12-sulfatide). When incubated at pH 5.0 in the presence of 5 mM MnCl2 and 5.5 mM of taurodeoxycholate, either substrate was hydrolyzed by arylsulfatase A of human leukocytes. The rate of hydrolysis was proportional to the incubation time and concentration of enzyme; Michaelis-Menten type kinetics were observed with increasing concentrations of substrate. For determining the rate of hydrolysis, each of the two products (i.e., P12- and PSA12-cerebrosides) were separated from the bulk of respective unreacted sulfatide on small columns of DEAE-Sephadex A-25 and their fluorescence intensities read at 343-378 and 350-380 nm for the excitation and emission wavelengths for P12- and PSA12-cerebrosides, respectively. When extracts of skin fibroblasts derived from normal individuals and patients with Maroteaux-Lamy (lacking arylsulfatase B) or metachromatic leukodystrophy (lacking arylsulfatase A) were used as source of enzyme, P12 sulfatide was hydrolyzed by the former two but not by the latter cell extract. Several derivatives of cerebroside sulfate were also synthesized and found to inhibit the hydrolysis of pyrenesulfatide by leukocyte arylsulfatase A. The results demonstrate that these two pyrene containing sulfatides can be effectively used as specific substrates for the determination of arylsulfatase A activity in extract of cells and most probably also of tissues. PMID- 2564283 TI - Multiple-dose pharmacokinetics of the new H1-receptor antagonist tazifylline in healthy volunteers. AB - The multiple-dose pharmacokinetics of the new H1-receptor antagonist, tazifylline, were investigated in healthy volunteers. From single-dose data, tazifylline appeared to be rapidly absorbed (median tmax of 0.6 h) and eliminated (t1/2 = 1.0 +/- 0.2 h). However, plasma levels measured on days 3 and 8 of the multiple-dose regimen (10 mg b.i.d. for 8 days) indicated moderate accumulation. A two-compartment model best described multiple-dose data with a terminal half life of 15.6 +/- 7.6 h consistent with twice-daily dosing of tazifylline. PMID- 2564284 TI - Blasted with ennui. PMID- 2564285 TI - Cardiotoxic effect with convulsions in terfenadine overdose. PMID- 2564286 TI - Optimum duration of antithyroid drug treatment determined by assay of thyroid stimulating antibody in patients with Graves' disease. AB - OBJECTIVE: To determine the optimal duration of antithyroid drug treatment by monitoring serum thyroid stimulating antibody values in patients with Graves' disease. DESIGN: Prospective longitudinal trial of patients with Graves' disease followed up for 24 months after withdrawal of treatment. SETTING: Tertiary referral centre. PATIENTS: A total of 64 consecutive patients with untreated Graves' disease, eight of whom were subsequently excluded. Fifty six patients completed the study. INTERVENTIONS: All patients were treated initially with carbimazole 40 mg, then with decreasing doses that maintained a euthyroid state. Treatment was scheduled to continue for 18 months but was withdrawn earlier if serum thyroid stimulating antibody became undetectable. END POINT: Serum values of thyroid stimulating antibody (assayed by stimulation of human thyroid cells in vitro) and thyroid hormones and thyroid state every three months during treatment and afterwards every six months for 24 months. MEASUREMENTS AND MAIN RESULTS: In 44 patients serum thyroid stimulating antibody became undetectable during treatment and treatment was withdrawn (median duration of treatment nine months, range 3-18 months). In 12 patients the antibody could be detected during 18 months of treatment. Among the first group of 44 patients initial values of the antibody before treatment were significantly lower than in the second group of 12 patients (median 225% (range 138-1236%) v 570% (250-1480%), p less than 0.001); the incidence of relapse was also lower (41% v 92%, p less than 0.001); and among those who did relapse the disease free interval after treatment was longer (median 12 months v 1 month, p less than 0.001). Moreover, the initial median serum values of thyroid stimulating antibodies were not related to the occurrence of relapse or remission as these did not differ between patients who did and did not have a relapse (median 267% (range 139-1480%) v 220% (range 138-1236%). CONCLUSION: Monitoring of serum thyroid stimulating antibody was a good guide to the duration of treatment as it allowed the treatment period to be considerably shortened in a large group of patients with no loss of efficiency. PMID- 2564287 TI - The optimization of collection of peripheral blood stem cells for autotransplantation in acute myeloid leukaemia. AB - Between November 1982 and November 1986 31 patients with acute myeloid leukaemia underwent peripheral blood stem cell apheresis during haemopoietic regeneration following induction chemotherapy. A retrospective analysis of the factors affecting the efficacy of stem cell harvest and of the clinical outcome of these patients was performed. The mean number of myeloid progenitor cells (CFU-GM) collected was significantly higher in the complete remission group (n = 22) than in the partial remission group (n = 9). Fifty x 10(4) CFU-GM/kg body weight or more, which produced rapid, complete and sustained haemopoietic reconstitution after autografting in our patients, were collected from six of nine patients who underwent three or four 7-litre aphereses over 5-7 days using a lymphocyte collection procedure on the Fenwal CS3000 [Protocol B] but only from two of 12 patients who underwent three or four 5-litre aphereses over 3-5 days using the Aminco Celltrifuge [Protocol A] (p less than 0.05). No adverse effects on the rates of neutrophil, platelet and lymphocyte recovery after induction chemotherapy or on long-term disease-free survival for patients who achieved a complete remission could be attributed to apheresis when compared with a historical control group of 39 patients who achieved complete remission following the same induction chemotherapy but did not undergo apheresis. We conclude that sufficient numbers of peripheral blood stem cells to produce safe and rapid haemopoietic reconstitution can be collected from most patients who achieve complete remission using apheresis Protocol B without impairment of haemopoietic recovery or adversely affecting the length of complete remission. PMID- 2564288 TI - The thermogenic actions of alpha 2-adrenoceptor agonists in reserpinized mice are mediated via a central postsynaptic alpha 2-adrenoceptor mechanism. AB - 1. The dose-related effects of the selective alpha 2-adrenoceptor agonists clonidine, UK-14,304 and B-HT 933 on the body temperature of untreated and reserpine-treated mice were investigated. 2. In untreated mice all three agonists induced a dose-related hypothermia. The highest doses of UK-14,304 and B-HT 933, 3 and 100 mg kg-1 respectively, elicited a marked (10 degrees C) hypothermia, whereas the maximal hypothermic effect of clonidine (5.5 degrees C) was less pronounced and reached a plateau at a dose of 0.5 mg kg-1 i.p. 3. Reserpine (2.5 mg kg-1, s.c.) induced a marked hypothermia in the mouse; 18 h after injection body temperature had decreased to only slightly (0.5-1.5 degrees C) above ambient (19 degrees C). 4. All three alpha 2-agonists produced a partial dose-related reversal of reserpine-induced hypothermia; maximal thermogenic responses (9-10 degrees C increases in body temperature) were elicited by doses of 0.2, 0.5 and 16 mg kg-1 i.p. of clonidine, UK-14,304 and B-HT 933 respectively, and the log dose-response curves for all 3 agonists were bell-shaped. 5. Following intracerebroventricular administration to reserpine-treated mice, the thermogenic response to clonidine was more rapid in onset, and the agonist was 20 fold more potent than when injected i.p. 6. The selective alpha 2-adrenoceptor antagonists, idazoxan (0.05-0.5 mg kg-1), Wy 26392 (0.3-5.0 mg kg-1) and yohimbine (0.1-1.6 mg kg-1) given orally attenuated the thermogenic responses to all 3 agonists in reserpinized mice in a dose-related manner. Pretreatment with a single dose of idazoxan (0.3 mg kg-1, orally) elicited a 6 fold parallel shift to the right in the dose-response curve to clonidine. 7. The selective alpha 1-adrenoceptor antagonists, prazosin (10 mg kg-1) and indoramin (3-10 mg kg-1), and the beta adrenoceptor antagonist, propranolol (10 mg kg-1), only partially attenuated the thermogenic responses to the alpha 2-agonists in reserpinized mice. These effects were variable and not clearly dose-related. 8. Pretreatment of reserpinized mice with the catecholamine synthesis inhibitor, alpha-methyl-p-tyrosine, markedly attenuated (60-95%) the thermogenic response to the noradrenaline uptake inhibitor, desipramine (0.13-12.5 mg kg-1, i.p.), but only slightly reduced (10 35%) that to clonidine (0.032-0.5 mg kg-1, i.p.). 9. These results suggest that alpha2-adrenoceptor agonists reverse reserpine-induced hypothermia via a central mechanism involving activation of postsynaptic alpha 2-adrenoceptors. PMID- 2564289 TI - The effects of acute and chronic desipramine on the thermogenic and hypoactivity responses to alpha 2-agonists in reserpinized and normal mice. AB - 1. The effects of acute and chronic (14 day) administration of the noradrenaline uptake inhibitor, desipramine (DMI), on the thermogenic responses to clonidine in reserpine-treated mice, and on the hypothermic and hypoactivity responses to the alpha 2-agonist, UK-14,304, in untreated mice were examined. 2. Taking the capacity of DMI to delay the onset of reserpine-induced hypothermia as an indicator of noradrenaline (NA) uptake inhibition, the lowest dose of DMI to inhibit uptake significantly for 12 h in the mouse was shown to be between 10 and 20 mg kg-1 orally. 3. Chronic (every 12 h for 14 days), but not acute treatment with DMI (15 mg kg-1, orally), attenuated the thermogenic responses to low doses (0.02-0.225 mg kg-1, i.p.) of clonidine (injected 20 h after the last dose of DMI) in reserpinized mice. 4. Acute DMI administration slightly attenuated the hypothermia and hypoactivity induced by UK-14,304 (0.25-1.0 mg kg-1, i.p.) when injected 2h, but not when injected 18-21h before the agonist. In contrast, 18-21h after withdrawal from chronic DMI both of these responses to UK-14, 304 were markedly attenuated. 5. As the thermogenic response to clonidine in reserpinized mice appears to involve central post-synaptic alpha 2-adrenoceptors, these results suggest that prolonged inhibition of NA uptake decreases the sensitivity of postsynaptic alpha 2-adrenoceptors. The results of the studies using UK-14,304 indicate that central alpha 2-adrenoceptors involved in mediating other behavioural and pharmacological responses to alpha 2-agonists are also down regulated by chronic inhibition of NA uptake. PMID- 2564290 TI - Mechanism of dexamphetamine-induced mydriasis in the anaesthetized rat. AB - 1. The effect of intravenous administration of dexamphetamine [+)-Amp) on rat pupil diameter was investigated. In all experiments, the vagosympathetic trunks were sectioned bilaterally at the cervical level. 2. In rats anaesthetized with urethane, (+)-Amp (0.1-0.3 mg kg-1, i.v.) produced a dose-related increase in pupil size. The mydriatic effects of (+)-Amp were evident immediately after administration. 3. Pretreatment with the alpha 2-adrenoceptor antagonists yohimbine (1.5 mg kg-1 i.v.) or idazoxan (0.5 mg kg-1, i.v.) blocked the pupillary response to (+)-Amp. Yohimbine caused about a 30 fold shift to the right in the dose-response curve whereas idazoxan almost completely abolished the mydriatic response to (+)-Amp. 4. In contrast, pretreatment with the alpha 1 adrenoceptor antagonist phenoxybenzamine (2 mg kg-1, i.v.), failed to alter significantly the pupillary response to (+)-Amp. 5. Depletion of central nervous system (CNS) monoamines with reserpine (5 mg kg-1, i.p.) and alpha-methyl-p tyrosine (2 x 300 mg kg-1, i.p.) prevented the pupillary response to (+)-Amp. 6. The mydriatic effect of (+)-Amp was present only in preparations that had intact parasympathetic neural tone to the iris. Central preganglionic denervation of the oculomotor nerve abolished the mydriatic response of (+)-Amp. 7. These results indicate the (+)-Amp acts in the CNS to produce mydriasis in the anaesthetized rat by stimulating CNS postsynaptic alpha 2-adrenoceptors, findings that are consistent with the hypothesis that (+)-Amp acts predominantly as an indirect sympathomimetic agent to release endogenous stores of a monoaminergic neurotransmitter (perhaps noradrenaline). PMID- 2564291 TI - Expression of functional postjunctional alpha 2-adrenoceptors in rabbit isolated distal saphenous artery--a permissive role for angiotensin II? AB - In the rabbit isolated distal saphenous artery, the population of postjunctional adrenoceptors is of the alpha 1 variety under normal in vitro experimental conditions, based on the potency order of selective agonists and on the effects of the antagonists prazosin and rauwolscine against responses to UK-14304. Angiotensin II (A II, 0.05 microM) however, without affecting resting baseline tension, markedly enhanced responses to UK-14304, particularly at low concentrations. This previously unseen component of the response to UK-14304 was resistant to prazosin (0.1 microM) but susceptible to rauwolscine (1 microM). A II would therefore appear to have a permissive role for the expression of a quiescent population of postjunctional alpha 2-adrenoceptors in the rabbit distal saphenous artery. PMID- 2564292 TI - A local circuit neocortical synapse that operates via both NMDA and non-NMDA receptors. AB - 1. In slices of rat neocortex, spike triggered averaging was employed to record in one neurone the excitatory postsynaptic potential (e.p.s.p.) generated by a spike in another, neighbouring neurone. When recorded at different membrane potentials, some of these e.p.s.ps exhibited a voltage relation typical of neuronal responses to N-methyl-D-aspartate (NMDA). 2. Selective NMDA antagonists reduced the amplitude of these e.p.s.ps, but had little effect on their early rising phase. In contrast, a less selective excitatory amino acid antagonist reduced all phases of the e.p.s.p. 3. By analyzing single axon e.p.s.ps we have been able to establish that the synaptic input to one cortical cell, delivered by a single presynaptic cortical cell, operates simultaneously via NMDA and non-NMDA amino acid receptors. PMID- 2564293 TI - Time course and extent of alpha 1-adrenoceptor density changes in rat heart after beta-adrenoceptor blockade. AB - 1. It has been suggested that impaired beta-adrenoceptor stimulation is a condition under which the functional role of cardiac alpha 1-adrenoceptors is enhanced. We therefore investigated the extent and time course of changes in alpha 1-adrenoceptor characteristics after chronic treatment with the beta adrenoceptor blocker propranolol in rat heart. For comparison beta-adrenoceptors were also studied. The mechanism of the changes in adrenoceptor density was investigated with cycloheximide, an inhibitor of protein synthesis. The functional significance of an increased alpha 1-adrenoceptor density was tested by measuring isometric force of contraction in the presence of phenylephrine or isoprenaline in right ventricular papillary muscles. 2. Rats were treated with propranolol (9.9 mg kg-1 daily) or 0.9% NaCl, applied with osmotic minipumps for 1, 2, 3 or 7 days. Propranolol treatment resulted in a maximally 28% increase of alpha 1-adrenoceptor density after 3 days (NaCl 95.9 +/- 3.5 vs. propranolol 123.0 +/- 1.6 fmol mg-1 protein, n = 6, P less than 0.01). This up regulation reached significant levels after 2 days of treatment and was reversible after cessation of treatment within two days. KD-values were the same for NaCl- and propranolol-treated rats. Changes of Bmax and KD in beta-adrenoceptor binding assays did not reach significant levels. 3. Cycloheximide (1.5 mg kg-1 i.p. daily for 3 days) inhibited the propranolol-induced increase in Bmax of alpha 1 adrenoceptors completely. In addition, cycloheximide also decreased the density of alpha 1- and beta-adrenoceptors also under control conditions. 4. pD2-values for the positive inotropic effect of phenylephrine and isoprenaline in isolated electrically driven papillary muscle were similar in NaCl- and propranolol treated rats (phenylephrine: 5.41 + 0.11 vs. 5.41 + 0.19, n = 7; isoprenaline: 6.31 + 0.18 vs. 6.65 + 0.19, n = 7). The observed increase in alpha-adrenoceptor density in healthy rat heart may therefore not be high enough to enhance the phenylephrine-induced increase in force of contraction. 5. In conclusion, time course and effects of cycloheximide indicate that the increase in B,,,, of myocardial alpha 1-adrenoceptors is due to de novo synthesis of receptors. However, at least for the rat heart model, a functional significance of this increase could not be demonstrated. PMID- 2564294 TI - Intravenous digital subtraction angiography in non-specific aorto-arteritis. AB - We have studied 40 patients with non-specific aorto-arteritis (Takayasu's disease) using intravenous digital subtraction angiography (DSA). The aorta, its major branches and (in 18 patients) pulmonary arteries were evaluated to determine the degree and extent of involvement. No complications related to the procedure were encountered. Good quality diagnostic images were obtained in 39 out of 43 instances. The unsuccessful examinations were in patients with congestive cardiac failure, and in one patient who would not co-operate. Aortic wall thickness and mild involvement of the descending aorta in the region of the diaphragm could not be assessed. Intravenous DSA is acceptable for the diagnosis and follow-up of aorto-arteritis in suitable patients. PMID- 2564295 TI - Tyrosine hydroxylase phosphorylation in rat brain striatal synaptosomes. AB - The present studies were carried out to determine if tyrosine hydroxylase phosphorylation in rat brain striatal synaptosomes is activated by dibutyryl cyclic AMP treatment. Incubation of synaptosomes with [32P]orthophosphate, followed by immunoprecipitation and sodium dodecyl sulfate polyacrylamide gel electrophoresis, produced a band of radioactivity associated with a 62 kDa polypeptide. Treatment with the catecholamine neurotoxin, 6-hydroxydopamine, produced parallel losses of: (1) tyrosine hydroxylase enzyme activity, (2) dopamine content, and (3) the 62 kDa band of radioactivity. These data support the identification of this band as a tyrosine hydroxylase-derived polypeptide. Incubation with dibutyryl cyclic AMP produced an increase in soluble tyrosine hydroxylase activity and phosphorylation. These results suggest that the increase in synaptosomal catecholamine synthesis produced by dibutyryl cyclic AMP is mediated by an increase in tyrosine hydroxylase phosphorylation. PMID- 2564296 TI - Evoked release of endogenous adenosine from rat cortical slices by K+ and glutamate. AB - Release of endogenous adenosine from rat cortical slices was determined in response to depolarization by 30 mM K+ and by exposure to glutamate. K+ and glutamate both released adenosine. Glutamate-evoked release was decreased by approximately 50% in the presence of the N-methyl-D-aspartate (NMDA) receptor antagonist, 2-amino-5-phosphonovaleric acid (APV). The adenosine released by glutamate could modulate neurotransmission, and may have a protective effect in pathologic conditions of excess excitation involving glutamate. PMID- 2564297 TI - A new group of tyrosine hydroxylase-immunoreactive neurons in the cat thalamus. AB - A new cell group composed of a large number of neurons immunoreactive to tyrosine hydroxylase (TH) was demonstrated in the paraventricular nucleus and midline nuclei of the cat thalamus, using four different anti-TH sera after colchicine treatment. However, in these regions, we did not detect any cell bodies containing other catecholamine synthesizing enzymes nor dopamine. PMID- 2564298 TI - Possible involvement of calcitonin gene-related peptide (CGRP) in non-cholinergic non-adrenergic relaxation induced by mesenteric nerve stimulation in guinea pig ileum. AB - Mesenteric nerve (MN) stimulation produced a biphasic response, i.e., a contraction followed by a prolonged relaxation in the isolated guinea pig ileum after complete adrenergic neuron blockade with guanethidine. This biphasic response mimicked the effect of capsaicin by itself. The latter relaxation response to MN stimulation was unaffected by hexamethonium, but was abolished by capsaicin in an irreversible fashion. Calcitonin gene-related peptide desensitization (CGRP-D) reduced the relaxation response to about 20% of the control. After pretreatment with atropine and guanethidine, MN stimulation provoked a pure relaxation, which was significantly reduced to about 20% of the control by CGRP-D. Therefore, it seems likely that the non-adrenergic non cholinergic relaxation response to MN stimulation is partly mediated via a release of CGRP from sensory nerve endings. PMID- 2564299 TI - Effects of ganglionic blocking agents on behavioral responses to centrally administered CRF. AB - The ganglionic blocking agents, chlorisondamine (CL) and hexamethonium (HEX) were used to examine the role of altered autonomic function in the behavioral response to i.c.v.-administered corticotropin-releasing factor (CRF). Animals were tested in either a novel modified open field or in their home cages. CRF-induced alterations in locomotion, grooming and eating were assessed in both environments in the presence or absence of CL or HEX. In the home cage the ability of CRF to increase grooming was attenuated by pretreatment with either CL or HEX. In the modified open field only HEX significantly suppressed grooming. In the familiar environment CRF-induced increases in locomotion were significantly inhibited by CL. However, in a novel environment, where CRF suppresses locomotion, CL was inactive. The competitive ganglionic nicotinic blocking agent, HEX, on the other hand, inhibited both the increased locomotion produced by CRF in the home cage and also the decreased locomotion induced by CRF in the modified open field. CRF suppression of food consumption was attenuated by CL. These results indicate that while centrally-mediated activation of the sympathetic nervous system cannot account for the full magnitude of the behavioral effects of i.c.v. CRF, such activation may play a part in both the locomotor activating components of the CRF response seen in the familiar environment as well as the suppressive effects seen in the novel environment. PMID- 2564300 TI - Age-related alterations in noradrenergic input to the hippocampal formation: structural and functional studies in intraocular transplants. AB - Intrinsic versus extrinsic determinants of age-related alterations in hippocampal noradrenergic transmission were investigated using intraocular allografts in rats. Three groups of animals were examined: young hippocampal transplants in young hosts, old transplants in old hosts and young transplants in old hosts. Postsynaptic sensitivity to noradrenaline (NA) was measured by extracellular recordings of spontaneous activity and superfusion with known concentrations of catecholamines in the anterior chamber of the eye. Hill plots demonstrated that the dose-response relationships of NA-induced depressions were linear and parallel in the 3 groups. Aged hippocampal grafts displayed a highly significant subsensitivity to NA of one order of magnitude. The EC50 for this group was 203.1 microM as compared to 29.2 in young grafts. Young intraocular grafts in old hosts responded similarly to transplants in young hosts, with an EC50 of 32.4 microM for the depressant actions of NA. Collaterals of the host iris sympathetic ground plexus invaded the hippocampal grafts. The density of this noradrenergic innervation was estimated by immunohistochemistry for tyrosine hydroxylase. A slightly increased density and fluorescence intensity of the noradrenergic fibers were observed in the old transplants as compared to the young transplants in young and old hosts. This was correlated with a significantly (P less than 0.01) increased content of NA in old transplants, as measured with high performance liquid chromatography. The old transplants also contained a large number of autofluorescent lipofuchsin granules, which were absent in the young transplants, regardless of the recipient age. Taken together, these results suggest the existence of alterations in pre- as well as postsynaptic noradrenergic mechanisms in the aging hippocampus. These changes were dependent on transplant age rather than host age, thus suggesting an involvement of intrinsic rather than extrinsic determinants in this model system. PMID- 2564301 TI - Excitatory and inhibitory neurotransmitters in the generation and degeneration of hippocampal neuroarchitecture. AB - The possibility that excitatory and inhibitory inputs to neurons can affect the generation and degeneration of neuroarchitecture was examined in hippocampal pyramidal neurons in isolated cell culture. Dendritic outgrowth and cell survival were directly monitored in neurons exposed to: the excitatory neurotransmitter glutamate, the inhibitory transmitter GABA, anticonvulsants or combinations of these agents. Glutamate caused a graded series of changes in pyramidal neuron cytoarchitecture: a selective inhibition in dendritic outgrowth and dendritic pruning was observed with subtoxic levels of glutamate while cell death was induced by higher levels. Low levels of GABA alone or in combination with diazepam, carbamazepine, phenobarbital or phenytoin were without effect on dendrite outgrowth while higher levels caused moderate reductions in outgrowth. Neither GABA nor the anticonvulsants affected cell survival. GABA plus diazepam, phenobarbital, carbamazepine and phenytoin each significantly reduced the dendritic regression and cell death normally caused by glutamate. Elevation of extracellular K+ to 50 mM caused dendritic regression and 100 mM K+ caused cell death; these effects were greatly reduced by GABA and anticonvulsants. The calcium channel blocker Co2+ prevented the dendritic regression and cell death caused by both glutamate and K+ indicating that calcium influx was required for the neuroarchitectural responses. Taken together, these results demonstrate that neurotransmitters and neuromodulatory drugs can have direct and interactive effects on both neurite outgrowth and cell survival. Such neurotransmitter actions may play roles in both the formation and degeneration of the neuronal circuits in which they participate in information coding. PMID- 2564302 TI - Effect of central administration of phencyclidine on plasma concentrations of luteinizing hormone. AB - Previous research has demonstrated that excitatory amino acids acting at N-methyl D-aspartate (NMDA) receptors stimulate the release of luteinizing hormone (LH). Castration also elevates LH, an effect that may also involve NMDA receptors since the specific NMDA antagonist, DL-2-amino-5-phosphonopentanoic acid (AP5), antagonizes this action. Since PCP antagonizes a variety of actions of NMDA agonists, we hypothesized that it would diminish the ability of glutamate, homocysteic acid and castration to elevate LH. Our data support this hypothesis. PMID- 2564303 TI - Differential sensitivity of dorsal and ventral root activity to magnesium and 2 amino-5-phosphonovalerate (APV) in an isolated mammalian spinal cord preparation. AB - Spontaneous activity emerging from the isolated hamster spinal cord simultaneously along the ventral and dorsal lumbar roots was shown by inter-spike interval analysis to have different firing patterns. Ventral root firing was reduced in solutions containing 1 mM magnesium or sub-micromolar concentrations of 2-amino-5-phosphonovalerate (APV), whereas the activity in the dorsal roots was unchanged. Differences in sensitivity to magnesium and APV were also shown on the evoked ventral and dorsal root reflexes, with the dorsal root reflex being unaffected, and the ventral root reflex exhibiting a long latency component that was sensitive to magnesium and APV. These results indicate that NMDA receptors are involved in the generation of part of the ventral root responses in the spinal cord, but not in the generation of the dorsal root reflex and spontaneous dorsal root activity. PMID- 2564304 TI - Steady-state levels of pro-dynorphin-related end-products from the brain of the amphibian, Xenopus laevis. AB - Steady-state analyses of prodynorphin-derived opioid peptides were conducted on acid extracts of the brain of the frog. Xenopus laevis. Radioimmunoassays specific for dynorphin A(1-17), dynorphin A(1-8), alpha-neoendorphin and dynorphin B coupled with gel filtration chromatography and reverse phase high performance liquid chromatography were used. The major prodynorphin-related end product detected was alpha-neoendorphin. Interestingly, Leu-enkephalin was also detected. Since the Xenopus proenkephalin precursor does not contain the Leu enkephalin sequence, these data suggest that some of the prodynorphin-related end products had been cleaved to yield Leu-enkephalin. PMID- 2564305 TI - Presence of catecholaminergic axon-terminals containing beta-adrenergic receptor in the periventricular zone of the rat hypothalamus. AB - The present study, using a light microscopic double-immunofluorescence method, has revealed the presence of fibers containing both tyrosine hydroxylase- and beta-adrenergic receptor-like immunoreactivities in the rat hypothalamic periventricular zone. Subsequent immunoelectron microscopic analysis demonstrated that these belong to axon terminals. These findings suggest that presynaptic beta adrenergic receptor is present in catecholaminergic terminals. PMID- 2564306 TI - N-methyl-D-aspartate antagonist applied to the spinal cord hindlimb enlargement reduces the amplitude of flexion reflex in the turtle. AB - APV (D(-)-2-amino-5-phosphonovalerate), an NMDA (N-methyl-D-aspartate) antagonist, was applied in situ onto segments of the hindlimb enlargement of the turtle spinal cord. APV reduced the response amplitude of the flexion reflex. In contrast, APV did not alter the responsiveness of the rostral scratch reflex. Afferents for the flexion reflex enter the spinal cord via the dorsal roots of the middle segment of the hindlimb enlargement; afferents for the rostral scratch reflex enter the spinal cord via dorsal roots located anterior to the hindlimb enlargement. The results are consistent with the hypothesis that sensory interneuron NMDA receptors, synaptically activated either directly or indirectly by nearby cutaneous afferent axons, play a role in the spinal cord processing of cutaneous information. PMID- 2564307 TI - Immunocytochemical evidence for direct connections between neurotensin-containing axons and dopaminergic neurons in the rat ventral midbrain tegmentum. AB - A light- and electron-microscopic double antigen localization technique was employed to study cellular relationships between neurotensin (NT)-containing axons and dopaminergic neurons in the substantia nigra and ventral tegmental area of the rat. Direct contacts were observed between NT-immunoreactive axon terminals and the dendrites and perikarya of neurons containing the dopamine biosynthetic enzyme, tyrosine hydroxylase (TH). NT-positive terminals were also found to contact unlabeled dendrites and neuronal somata. These results reveal an ultrastructural substrate through which endogenous NT might influence the activity of dopaminergic neurons in the ventral midbrain tegmentum. PMID- 2564308 TI - Sipple syndrome. PMID- 2564309 TI - [Hemodynamic effects of anesthesia using propofol, fentanyl, nitrous oxide and vecuronium in kidney transplantation in the adult]. PMID- 2564310 TI - The pharmacotherapy of septic shock. AB - Septic shock is a common clinical problem in the intensive care setting. The high mortality rate associated with septic shock, regardless of age, reflects the inadequacy of available therapeutic approaches. The purpose of this article is to review the current pharmacologic approaches to the treatment of septic shock with an emphasis on the pathophysiologic correlations of these treatments. PMID- 2564311 TI - Increased P-glycoprotein expression and multidrug-resistant gene (mdr1) amplification are infrequently found in fresh acute leukemia cells. Sequential analysis of 15 cases at initial presentation and relapsed stage. AB - Using a DNA probe of mdr1 and an anti-P-glycoprotein monoclonal antibody (MRK16), the authors investigated 19 cases of adult acute leukemia patients (one M1, six M2, three M3, one M4, three M5, two L1, and three L2), comparing leukemia cells at the initial presentation (I) with those at the relapsed stage (R). By Southern hybridization analysis mdr1 DNA levels were not amplified in 32 samples from 19 patients (I: 14, R: 18). By Northern hybridization analysis mdr1 mRNA levels were not expressed in ten samples from seven patients (I: 4, R: 6). By indirect immunofluorescent assay with MRK16 antibody P-glycoprotein was not detected in 30 samples from 18 patients (I: 13, R: 17). Thus, P-glycoprotein expression and mdr1 gene amplification occurred infrequently not only in leukemia cells at the initial presentation but also in those at the relapsed cases and may not be a major cause of refractoriness to antileukemia drugs in adult acute leukemia. PMID- 2564312 TI - Treatment of N-nitrosobis(2-oxopropyl)amine-induced pancreatic cancer in Syrian golden hamsters with D-Trp-6-LH-RH and somatostatin analogue RC-160 microcapsules. AB - Antitumoral effects of the agonist of luteinizing hormone-releasing hormone (D Trp-6-LH-RH) and the somatostatin analog RC-160 (D-Phe-Cys-Tyr-D-Trp-Lys-Val-Cys Trp-NH2) on chemically induced ductal pancreatic adenocarcinomas were studied. The tumors were induced in female Syrian golden hamsters by weekly s.c. injections of N-nitrosobis(2-oxopropyl)amine at a dose of 10 mg/kg b.w. for 6 weeks. 18 weeks after the last injection, the peptides in controlled-release microcapsule formulations were administered s.c. The animals received the following therapies: Group 1 (N = 15), vehicle only; Group 2 (N = 13), D-Trp-6-LH RH microcapsules releasing 25 micrograms/day injected s.c. once a month; Group 3 (N = 14), RC-160 microcapsules, liberating 25 micrograms/day administered s.c. every 15 days; Group 4 (N = 14), the combination of D-Trp-6-LH-RH plus RC-160 microcapsules. The experiment was terminated on the 80th day when all hamsters in the control group were dead, but in the treated Groups 2, 3, and 4, we observed 71, 77, and 86% of survival rate, respectively. In addition to the prolongation of survival, the combination treatment resulted in a significant decrease in the tumorous pancreatic weight, increase in the body weight of the animals, reduction in ascites from 100 to 8.3% and regressive histological changes in 67% of the specimens. Our findings suggest that somatostatin analogues and D-Trp-6-LH-RH could be considered for the development of hormonal therapy for pancreatic cancer. PMID- 2564313 TI - Inhibition of promotion and persistent nodule growth by S-adenosyl-L-methionine in rat liver carcinogenesis: role of remodeling and apoptosis. AB - The resistant hepatocyte model (initiation/selection) and the triphasic model (initiation/selection followed by phenobarbital, for a maximum of 16 weeks) were compared for their ability to generate enzyme-altered foci (EAF) and nodules in the liver of Wistar rats initiated by diethylnitrosamine. The effects of S adenosyl-L-methionine (SAM) on the development of preneoplastic tissue was tested in these experimental models. In the absence of phenobarbital (PB), EAF and early nodules (EN) went through a phase of rapid growth, between 4 and 9 weeks after initiation, to a phase in which progressive decrease in number and size occurred. By the 26th week only a few remodeling EAF and nodules were found. In PB-treated rats a rapid increase in the percentage of liver occupied by EAF and EN, up to the 9th week after initiation, was followed by a period of slow growth (from the 9th to the 20th week) and then, after PB withdrawal (20th week), by a drop in the number and size of EAF and EN. However, at the 26th week actively growing nodules with a low tendency to spontaneous remodeling (persistent nodules) developed. EAF and EN showed a high DNA synthesis 5 weeks after initiation. Thereafter, progressive decline in DNA synthesis, coupled with remodeling and decrease in number of biochemical markers, was seen both in the absence and, even though to a lesser extent, in the presence of PB, indicating that preneoplastic lesions became increasingly insensitive to PB. Relatively few apoptotic bodies could be observed in EAF and EN during PB treatment. After PB withdrawal, decrease in growth potential was coupled with increase in apoptotic bodies. In contrast, in persistent nodules relatively high apoptosis occurred which partially counterbalanced high DNA synthesis. Administration of SAM for a maximum of 16 weeks, starting at the 4th week after initiation, caused a great decrease in number and size of EAF and EN, associated with inhibition of DNA synthesis, high cell death by apoptosis, high remodeling, and loss of biochemical markers, in preneoplastic lesions of both PB-treated and untreated rats. A 1-8-week SAM treatment, started after the development of persistent nodules, caused a great regression of nodular lesions, coupled with a sharp fall in DNA synthesis and increase in apoptosis. It is suggested that inhibition by SAM of the development of preneoplastic tissue is linked to a shift of the equilibrium between cell production and cell death in favor of cell death.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2564314 TI - Characterization of rat T cell precursors sorted by chemotactic migration toward thymotaxin. AB - An established rat thymic cell line secretes a peptide in the 11 kd range called thymotaxin that attracts a small subset of juvenile rat bone marrow cells via a chemotactic mechanism. The selected cell subset (0.1% of the total bone marrow) is composed of low-density lymphoid cells that do not replicate, and display an immature Thy-1+T-B- phenotype. Thymotaxin-responding cells do not grow in semi solid cultures under hemopoietic growth factors stimulation, and survive only in coculture with thymic stroma under steroid-free conditions. This stroma mimics the thymic microenvironment and allows a fraction of responding bone marrow cells to acquire T cell differentiation markers and to synthesize transcripts of the TCR alpha and beta chains. Chemotactic migration toward thymic epithelial cell peptides can be used in vitro to sort pre-T cells from the rat bone marrow. The sorted T cell precursors are resting stem cells possibly committed to lymphoid lineage. PMID- 2564315 TI - Translation of cyclin mRNA is necessary for extracts of activated xenopus eggs to enter mitosis. AB - The cyclins are a family of proteins encoded by maternal mRNA. Cyclin polypeptides accumulate during interphase and are destroyed during mitosis at about the time of entry into anaphase. We show here that Xenopus oocytes contain mRNAs encoding two cyclins that are major translation products in a cell-free extract from activated eggs. Cutting these mRNAs with antisense oligonucleotides and endogenous RNAase H blocks entry into mitosis in a cell-free egg extract. The extracts can enter mitosis if either of the cyclin mRNAs is left intact. We conclude that the synthesis of these cyclins is necessary for mitotic cell cycles in cleaving Xenopus embryos. PMID- 2564317 TI - Human CD2 3'-flanking sequences confer high-level, T cell-specific, position independent gene expression in transgenic mice. AB - We have localized a set of T cell-specific DNAase I hypersensitive sites in the 3'-flanking region of the human CD2 gene. A 5.5 kb BamHI-XbaI fragment containing these DNAase I hypersensitive sites conferred efficient, copy number-dependent, T cell-specific expression of a linked human CD2 minigene, independent of the position of integration in the transgenic mouse genome. When linked to the mouse Thy-1.1 gene or the human beta-globin gene, this fragment conferred the same T cell-specific expression, independent of its orientation. These results suggest that this flanking region is both necessary and sufficient for full tissue specific activation of homologous and heterologous genes in transgenic mice. PMID- 2564316 TI - Expression and function of Drosophila cyclin A during embryonic cell cycle progression. AB - Cyclin proteins are thought to trigger entry into mitosis. During mitosis they are rapidly degraded. Therefore, mitosis and consequently cyclin degradation might be triggered at a time when cyclins have reaccumulated to a critical level. We cloned and sequenced a Drosophila cyclin A homolog and identified mutations in the corresponding gene. Immunofluorescent staining revealed that cyclin A accumulates in the interphase cytoplasm of cellularized embryos, but relocates to the nuclear region early in prophase and is completely degraded within metaphase. Cyclin A was expressed in dividing cells throughout development, and a functional cyclin A gene was required for continued division after exhaustion of maternally contributed cyclin A. Importantly, the timing of post cellularization divisions was not governed by the rate of accumulation or level of cyclin A. PMID- 2564318 TI - [Anxiolytics 1985-1986]. PMID- 2564319 TI - Mechanistic studies on the DDT-induced inhibition of intercellular communication. AB - Two structurally unrelated compounds, 1,1'-(2,2,2-trichloroethylidene) bis(4 chlorobenzene) (DDT) and 12-O-tetradecanoylphorbol-13-acetate (TPA), are both potent inhibitors of cell-cell communication in vitro as well as tumour promoters in vivo. There is evidence that TPA acts via a specific receptor mechanism involving activation of protein kinase C (pkC). The mechanism of action of DDT has been discussed in terms of membrane perturbation, increased intracellular calcium, interaction with calmodulin and decreased cAMP levels. In the present study the objective was to examine the potential role of pkC activation in DDT induced inhibition of intercellular communication in cultured cells. The V79 metabolic cooperation assay was used for measuring intercellular communication. Furthermore, the effects of DDT on the activity of partially purified pkC from V79 cells was measured, as was the interaction of DDT with the phorbol ester/DAG binding site on the pkC enzyme. Results from the biochemical studies showed that DDT neither activates pkC nor binds to the phorbol ester/DAG-binding site, as measured by displacement of PDBU binding. Using the metabolic cooperation assay it was demonstrated that pretreatment with TPA made cells refractory, i.e. a second application of TPA did not inhibit cell-cell communication. DDT added to cells down-regulated with TPA inhibited cell-cell communication, even though these cells were refractive to TPA. This result further supports the hypothesis that DDT and TPA inhibit intercellular communication primarily by different pathways. At non-cytotoxic concentrations, pkC inhibitors (H7, W7 and palmitoyl carnitine) did not affect the TPA- or DDT-induced inhibition of cell-cell communication in the V79 metabolic cooperation assay. Quercetin, a pkC inhibitor which has been reported to eliminate DDT- or TPA-induced inhibition of intercellular communication, was investigated in an in vivo study that measured promotion of enzyme-altered foci in DEN-treated rat liver. Quercetin co administered with DDT did not act as an antipromoter. PMID- 2564320 TI - Differential susceptibility of a fish, tilapia Oreochromis mossambicus (Teleostei, Cichlidae) to hepatocarcinogenesis by diethylnitrosamine and methylazoxymethanol acetate. AB - Oreochromis mossambicus, commonly known as tilapia, is a freshwater teleost with a wide tolerance to environmental conditions. Multiple focal lesions in the liver were observed 2 months after cessation of a one-month long treatment with 100 p.p.m. diethylnitrosamine. Cells were small and compact and arranged in sheets. Ultrastructurally, these cells have abundant endoplasmic reticulum, round mitochondria, less conspicuous golgi apparatus and fat droplets. Other organs like the intestines, spleen, kidneys, ovaries and pituitary appeared normal. Two liver inducible enzymes, gamma-glutamyl transferase and tyrosine aminotransferase were elevated by 5- and 3-fold respectively. Aggressive migration of hepatocytes was observed in tumorigenic liver explants. Vitellogenesis and early embryological development appeared unaffected as the female fish spawned during hepatocarcinogenesis. However, their fry were stunted and short-lived. To compare the susceptibility of tilapia to another hepatocarcinogen, the fish were also treated with methylazoxymethanol acetate at 10 p.p.m. for 0.5-1 h. However, methylazoxymethanol acetate was too toxic and 75% of the fish succumbed 1 day after treatment. Moreover, after 2 months post-treatment, neither tumors nor change in enzyme activities were observed in any organ. These results suggest that tilapia could be a useful model for screening and differentiating carcinogens since they could develop liver tumors within only 2 months after treatment with diethylnitrosamine. PMID- 2564321 TI - Centrifugal elutriation of hepatocytes from 2-acetylaminofluorene-treated rats and their characterization by flow cytometry. AB - Treatment of male Wistar rats with 2-acetylaminofluorene (2-AAF) markedly altered the ploidy distribution of liver cells. Small diploid hepatocytes first appeared after 4-5 weeks feeding of a diet containing 0.02% 2-AAF; after 9 weeks 65-70% of the hepatocytes were diploid. Approximately two-thirds of this new liver cell population persisted after termination of the treatment. The hepatocytes from 2 AAF treated animals were separated according to size and ploidy by centrifugal elutriation and stained for gamma-glutamyltranspeptidase (gamma-GTase). The percentage of gamma-GTase-positive hepatocytes did not significantly differ between the various elutriated cell fractions. Thus gamma-GTase-positive liver cells obtained by feeding of 2-AAF do not represent a distinct size class of hepatocytes. The significance of carcinogen-induced diploid hepatocytes in hepatocarcinogenesis is discussed. PMID- 2564322 TI - The role of GSH depletion and toxicity in hydroquinone-induced development of enzyme-altered foci. AB - Hydroquinone (HQ) may activate oxygen via redox cycles in biological systems and may also deplete glutathione (GSH). Both these reactions are potentially harmful, and we have studied their possible involvement in hydroquinone-induced development of gamma-glutamyltranspeptidase (GGT)-positive enzyme-altered foci in rat liver. The effect of HQ was compared to the effect of duroquinone, catechol, resorcinol and phenol. The dose was 100 mg/kg per day and the test substances were administered for 7-12 weeks in these foci experiments. HQ gave an increased number of foci and increased the foci volume, while none of the other compounds had any significant effect on these parameters. HQ, duroquinone and resorcinol were also tested at a higher dose level (200 mg/kg per day), but this dose gave a lower number of foci than the 100-mg dose. HQ, duroquinone and catechol induced single-strand breaks in hepatic DNA. Single doses of HQ (200 mg/kg) increased malondialdehyde excretion in urine, indicating in vivo lipid peroxidation. Duroquinone, phenol and resorcinol were negative with respect to malondialdehyde excretion. Catechol could not be properly tested as the 200-mg dose killed several animals. HQ and catechol induced hepatic ornithine decarboxylase activity. This effect was correlated to GSH depletion. An in vitro model for toxicity studies with hepatocytes from carcinogen-treated rats was also used. In this model HQ could be shown to be selectively toxic to GGT-negative cells in the presence of extracellular GSH. The toxicity was preceded by a rapid depletion of GSH. Catechol also depleted GSH and could be shown to be selectively toxic, but higher concentrations than those used for HQ had to be used. Duroquinone, phenol and resorcinol were not selectively toxic to GGT-negative cells. As duroquinone can be regarded as a more potent inducer of redox cycles than HQ, it can be concluded that the foci data provide no evidence for an involvement of redox cycles in HQ induced development of enzyme-altered foci. They suggest that GSH depletion may act to develop enzyme-altered foci, and the in vitro data indicate a mechanism by which GSH depletion and toxicity may induce this effect. PMID- 2564323 TI - Effects of regional alpha- and beta-blockade on resting and hyperemic coronary blood flow in conscious, unstressed humans. AB - Our purpose was to determine if there are basal adrenergic influences on the coronary circulation in humans. We studied 56 patients with denervated hearts after cardiac transplantation and 19 normally innervated patients with angiographically normal coronary arteries. Coronary blood flow velocity was measured during cardiac catheterization with a subselective 3F intracoronary Doppler catheter. Heart rate was controlled by atrial pacing. Epicardial coronary artery diameter was measured by automated analysis of digital coronary angiograms. Coronary flow reserve was assessed by intracoronary papaverine hydrochloride (12 mg) injections. Regional sympathetic blockade was produced by intracoronary injections of phentolamine (3 mg, alpha) and propranolol (2 mg, beta) or metoprolol (3 mg, beta 1). After alpha-blockade, mean arterial pressure fell significantly (p less than 0.05) in both the denervated transplant (-5.8 +/- 1.5%) (mean +/- SEM) and normally innervated patients (-12.6 +/- 3.2%). Reductions in coronary flow velocity also were observed in these groups (-8.2 +/- 2.3% and -9.2 +/- 5.8%, respectively). Calculated coronary vascular resistance was unchanged. Similar changes were seen when patients were pretreated with beta blockade before alpha-blockade. Nonspecific beta-blockade did not affect mean arterial pressure but decreased coronary velocity (innervated, -11.6 +/- 3.9%; denervated, -9.3 +/- 2.4%) and increased coronary vascular resistance (innervated, 15.4 +/- 6.7%; denervated, 10.2 +/- 3.7%). Coronary vascular resistance did not rise in either group after selective beta 1-blockade with metoprolol. Coronary flow reserve did not change in either patient group after either alpha- or beta-blockade. Changes in epicardial coronary artery diameter were small and generally not significant. These data suggest that alpha-receptor mediated vascular tone is negligible in both denervated transplant patients and normally innervated patients. Additionally, the increase in vascular resistance after nonselective beta-blockade is the result of direct beta 2 vascular effects. Our data further suggest that there is little adrenergically mediated epicardial artery tone (either humoral or neural) at rest and that maximal vasodilator responses are not limited by adrenergically mediated vasomotor tone. PMID- 2564324 TI - Effects of verapamil and propranolol on changes in extracellular K+, pH, and local activation during graded coronary flow in the pig. AB - The beta-adrenergic and calcium channel blocking agents are known to reduce heart rate and alter myocardial contractility. More recent evidence suggests that both agents affect the metabolic consequences of ischemia, independent of their effects on heart rate and contractility. We used a low-flow model of ischemia in swine with heart rate held constant by atrial pacing. Blood was shunted from the carotid artery to the left anterior descending coronary artery through a controlled-flow roller pump to assess the threshold flow for the rise in extracellular potassium ([K+]e) and fall in extracellular pH (pHe) associated with ischemia during control situations and after the administration of either propranolol or verapamil. We also measured the changes in activation delay and contractility associated with graded flow reductions in the presence and absence of these drugs. We found that when heart rate is held constant, 1) verapamil shifts the threshold flow for [K+]e and pHe to lower levels, but propranolol does not; 2) verapamil lessens activation delay, while propranolol aggravates the delay; and 3) verapamil reduces afterload and selectively depresses contractility in the reperfused ischemic zone. We conclude that the calcium channel blockers and the beta-adrenergic-blocking agents have different effects and possibly different modes of action and should not be considered interchangeable when evaluating therapeutic options for patients with ischemic heart disease. PMID- 2564325 TI - Female haemophilia A in a family with seeming extreme bidirectional lyonization tendency: abnormal premature X-chromosome inactivation? AB - We studied a female child with mild classical haemophilia A, presenting with a F VIII deficiency similar to that detected in her maternal grandfather. Investigations on several occasions showed that the obligate carrier mother of the proposita had normal VIII:C activity, whereas her likewise obligate carrier sister had a typical carrier VIII:C/vWf:Ag pattern. The child was a phenotypically normal female with normal karyotype. Her father had no clinical or biochemical signs of haemophilia A. RFLP-analysis using DX13 and St14 probes each elicited one allele (5.8 and 3.4 kb, respectively) segregating along with the affected F VIII gene from the hemizygous grandfather to both his daughters and further to the haemophilic female child. The paternity of the child was analyzed using various red cell and HLA antigens and RFLP by p29C, a probe detecting polymorphic hypervariable TaqI and PstI fragments in the pseudoautosomal areas of the X- and Y-chromosomes. All results obtained were concordant with the declared paternity. RFLP-analysis, using single (Pst I) and double digestion (Pst I/Hha I) of DNA and a PGK probe, revealed a remarkable difference in hybridization fragments, strongly suggesting hypermethylation, and in consequence, preferential X-chromosome inactivation in the proposita. This points to extreme lyonization as the most plausible explanation for haemophilia A in this female child. A familial tendency to abnormal premature X-chromosome inactivation is speculated. PMID- 2564326 TI - DNA polymorphism at the locus for human cholesteryl ester transfer protein (CETP) is associated with high density lipoprotein cholesterol and apolipoprotein levels. AB - Cholesteryl ester transfer protein (CETP) is a protein involved in "reverse cholesterol transport" and it could play an important role in facilitating the removal of cholesteryl esters from peripheral tissues for transport to the liver or for transfer of cholesterol between plasma lipoprotein particles. Both functions may be relevant to susceptibility or resistance to atherosclerotic disease. We have studied 149 and 146 unrelated persons, respectively, for the A and B polymorphism at the CETP locus detectable with the restriction enzyme TaqI. The B system is by far the more polymorphic. A search for association with risk or "anti-risk" factor levels was conducted with the following quantitative parameters: total cholesterol, HDL cholesterol, triglycerides, apolipoprotein AI (apoA-I), apolipoprotein B (apoB) and Lp(a) lipoprotein levels. Highly significant differences in apoA-I concentration were found between the two categories of homozygotes in the B polymorphism. The association observed remained significant after multiplying the p value by the number of quantitative parameters used for the association tests. There was a dosage effect on the apoA I level of genes in the B polymorphism. We conclude that the associations observed are likely to reflect true biological phenomena. The effect of CETP genes appeared to be limited to non-smokers. PMID- 2564327 TI - Molecular Xp deletion in a male: suggestion of a locus for hypogonadotropic hypogonadism distal to the glycerol kinase and adrenal hypoplasia loci. AB - We have analyzed one patient with a syndrome of glycerol kinase deficiency (GKD), adrenal hypoplasia (AH), mental retardation (MR) and hypogonadotropic hypogonadism (HH). Although a cytogenetic analysis of the patient failed to reveal any detectable chromosomal abnormality, Southern blot analysis, using DNA probes from the Xp21-Xp22 region, revealed a molecular deletion localized between the DXS41 and the DXS268 loci. Our results together with those of others (van Ommen et al. 1986, 1987, Francke et al. 1987, Yates et al. 1987, Chelly et al. 1988) suggest that the GK gene is located between the DXS68 and DXS268 loci. In addition, we propose a locus for HH in Xp, distal to the genes for GK and AH. PMID- 2564328 TI - Resistance to experimental autoimmune thyroiditis: L3T4+ cells as mediators of both thyroglobulin-activated and TSH-induced suppression. AB - Mechanisms suppressive to induction of murine experimental autoimmune thyroiditis (EAT) can be activated by pretreatment with tolerogenic doses of mouse thyroglobulin (MTg) or prior TSH infusion to raise circulatory MTg levels. MTg activated suppressor T cells (Ts), shown earlier to be Thy-1+ and probably I-J+, were further characterized by in vivo administration of paired rat monoclonal antibodies to distinct epitopes on the L3T4 or Lyt-2 molecule, either on the day of, or subsequent to, initiation of the tolerogenic regimes. The cells required at the time of MTg pretreatment were L3T4+, Lyt-2- and low anti-L3T4 doses had no effect on their activation. The cells that mediated the strong MTg-induced resistance following pretreatment were also L3T4+; their suppressor function could only be abrogated by depletion of L3T4+, but not Lyt-2+, cells. Injection of cyclophosphamide (20-100 mg/kg) either prior to EAT induction or after Ts activation did not affect the severity of disease. Similarly, the suppressor state evoked by TSH infusion could only be abrogated by anti-L3T4 treatment. These findings indicate that both MTg-activated and TSH-induced suppression are mediated by L3T4+ cells. We hypothesize that MTg-specific Ts are present in normal, EAT-susceptible mice in low numbers to contribute to the maintenance of self-tolerance and that they are stimulated by increased levels of circulatory MTg to expand/differentiate and mediate the marked resistance to EAT induction. PMID- 2564329 TI - Study of DNA polymorphisms of the apolipoprotein AI-CIII-AIV gene cluster in patients with peripheral arterial disease. AB - 1. We have determined the frequency of DNA polymorphisms of the human apolipoprotein AI-CIII-AIV gene cluster, detected with XmnI, PstI, and PvuII, in a group of patients with peripheral arterial disease. 2. Of the patients, 81 had no evidence of disease in the coronary and carotid arteries, 73 had coronary artery disease but no evidence of carotid artery disease, 25 patients had carotid artery disease but no evidence of coronary artery disease, and 38 had both coronary and carotid artery disease. 3. Levels of triacylglycerol, cholesterol, apolipoprotein B and apolipoprotein AI were not significantly different between the four patient groups. 4. The frequencies of the alleles for the apolipoprotein AI-CIII-AIV polymorphisms, detected with XmnI, PstI and PvuII, did not differ significantly in the patient groups when compared with a sample of clinically well normolipidaemic individuals also from a London population. 5. All five patients with the XmnI genotype we designate X2X2 had high levels of cholesterol, apolipoprotein B and apolipoprotein AI. 6. Patients with the rare VB2 allele of the apolipoprotein CIII-AIV restriction fragment length polymorphism had lower levels of cholesterol, acylglycerol and significantly lower levels of serum apolipoprotein. 7. Our observations suggest that variation in the apolipoprotein AI-CIII-AIV gene cluster may not be contributing significantly to the development of peripheral arterial disease, but variation associated with some of the restriction fragment length polymorphisms may be involved in determining levels of cholesterol- and apolipoprotein-B-containing lipoproteins. PMID- 2564330 TI - Why must some schizophrenic patients be involuntarily committed? The role of insight. AB - Twenty-four of 52 (46%) schizophrenic patients hospitalized because of acute psychotic episodes associated with preadmission medication noncompliance required involuntary commitment. Committed patients were rated as significantly more severely ill than voluntary patients and were significantly more likely to be transferred to extended treatment facilities after acute care. However, committed patients were significantly less likely than were voluntarily admitted patients to acknowledge that they were psychiatrically ill and in need of treatment, i.e., to demonstrate insight. Although psychopathology diminished significantly in both committed and voluntary patients over the course of hospitalization, only in voluntary patients did insight increase significantly. Over a 21/2 to 31/2 year follow-up, those patients who had been involuntarily committed at the index hospitalization were significantly more likely to require involuntary admissions than were the initially voluntary patients. Inability to see the self as ill seems to be a persistent trait in some schizophrenic patients. PMID- 2564331 TI - [Angioplasty of the subclavian artery. The technic, early and late results]. AB - Percutaneous transluminal angioplasty (PTA) was performed in 22 patients (13 women and nine men; mean age 57 years [44-74] ) with symptomatic obstruction of the subclavian artery which was due to arteriosclerotic stenosis in 14, occlusion in six patients, radiogenic multiple stenoses in one (after radiotherapy for breast cancer), and stenoses of the left and right subclavian artery in one patient with Takayasu's arteritis. Primary success occurred in 21 (91%). The only complication was a stenosis of the brachial artery after combined brachiofemoral recanalization of a subclavian artery occlusion, but it did not require treatment. No emboli were noted. A good long-term result was achieved in 19 of the 21 primarily successful PTAs, in two instances after repeat PTA. PTA has thus been shown to be a simple, well tolerated interventional treatment of symptomatic subclavian obstruction with a high success rate and low risk of complications. It is to be preferred as a primary procedure to surgical intervention. PMID- 2564332 TI - [Drug therapy of unstable angina pectoris]. PMID- 2564333 TI - [Gene technology in animal husbandry, new research approaches exemplified by the milk protein gene of cattle]. AB - For investigation of bovine milk protein genes several methods of recombinant DNA techniques are presented. Possible applications of genome research in animal breeding are given including the characterization of structure and function for single genes, gene mapping as well as screening for gene variants in populations. Hence it follows that scientific and practical developments can be expected and will be an influence on future animal production. PMID- 2564334 TI - [Diagnosis of hereditary diseases with molecular biology methods]. AB - The identification of phenotypically normal carriers of genetic defects is crucial for eliminating recessive defect genes from farm animal populations. Recombinant DNA techniques allow us to identify defect genes on the DNA-basis regardless of the animals age and time of gene expression. Genetic defects also are important as model systems to investigate the regulation of metabolic pathways and the mechanisms of embryonic development. PMID- 2564335 TI - [Standardization of ICO-G2 monoclonal antibodies against X-hapten]. AB - Results obtained from comparative analysis of ICO-G2 monoclonal antibody with some standard CD 15 monoclonal antibodies (FMC-11, TG-1, G-2, B-3-4) and with MG 1, MG-3, MG-5, MG-6 monoclonal antibodies (Prague) performed by flow cytometry on FACS 440 instrument (Becton Dickinson) indicate that all monoclonal antibodies have a similar reactivity and may identify the same X-haptene structure (also known as Lex) included into the differentiation cluster as CD15. PMID- 2564336 TI - Sodium butyrate induces pyroglutamyl peptidase I and decreases thyrotropin releasing hormone receptors in GH3 cells. AB - The effect of sodium butyrate treatment on TRH-degrading enzymes and TRH receptors in GH3 cells was investigated. The specific activity of pyroglutamyl peptidase I (EC 3.4.19.3) was increased by exposure to sodium butyrate in a time- and concentration-dependent manner, whereas the specific activity of prolyl endopeptidase (EC 3.4.21.26) was unchanged. The maximal effect occurred at a concentration of 1 mM sodium butyrate and 16 h after exposure. The increase was reversible upon removal of sodium butyrate from the cell culture. Cycloheximide totally blocked the stimulation, indicating that the increase was due to new protein synthesis. Sodium butyrate had no effect on pyroglutamyl peptidase I activity in the AtT-20 cell line. [methyl-3H]TRH binding to intact GH3 cells was reduced to 70% of the control value when cells were exposed to 1 mM sodium butyrate for 8 h. A maximal decrease in binding to 40% of the control value occurred after 16 h of exposure. The Kd of [methyl-3H]TRH binding was not changed. Sodium butyrate altered GH3 cell morphology, but the morphological changes occurred after alterations of pyroglutamyl peptidase I activity and [methyl-3H]TRH-binding sites. Other agents known to alter GH3 cell morphology had no effect on pyroglutamyl peptidase I activity. These results indicate that sodium butyrate can in some respects mimic the action of T3 on GH3 cells. Moreover, they provide further evidence that the activity of pyroglutamyl peptidase I, but not prolyl endopeptidase, is subject to regulation in the GH3 cell. PMID- 2564337 TI - In vivo and in vitro production of inhibin by cryptorchid testes from adult rats. AB - Adult male rats were made bilaterally cryptorchid for 3, 7, 10, and 14 days, and the peripheral serum, testicular vein serum, and interstitial fluid levels of inhibin were measured by RIA, and compared with values obtained in intact animals. The levels of inhibin in peripheral serum, testicular vein, and interstitial fluid were significantly elevated (P less than 0.01) after 3 days of cryptorchidism but declined significantly thereafter. The secretion of inhibin was also studied in vitro using isolated segments of seminiferous tubules from scrotal and abdominal testis in adult rats made unilaterally cryptorchid for 3, 6, and 12 days. The basal inhibin secretion by 3-day cryptorchid seminiferous tubules incubated at 37 C was significantly greater when compared with the scrotal seminiferous tubules at 32 C. If seminiferous tubules from scrotal testes were incubated at 37 C, the secretion of inhibin was greatly increased to similar levels observed by the 3-day abdominal seminiferous tubule cultures. In addition inhibin secretion was significantly higher when tubules from 5-week hypophysectomized rats were cultured at 37 C compared to 32 C. The inhibin secretion by seminiferous tubules from 6-day abdominal testes had returned to scrotal seminiferous tubule levels but then decreased below scrotal seminiferous tubule levels after 12 days of cryptorchidism. Seminiferous tubules from cryptorchid testes remain responsive to FSH stimulation (500 ng/ml) up to 12 days of cryptorchidism. FSH-stimulated inhibin production was increased at 3 and 12 days after cryptorchidism, but similar at 6 days after cryptorchidism, compared to the response of tubules obtained from scrotal testes. Furthermore, using seminiferous tubules from normal adult rats, FSH-stimulated inhibin production was increased by raising the incubation temperature from 32 C to 37 C. These in vivo and in vitro data suggest a dual stimulatory and inhibitory effect of higher temperature on inhibin production with an initial rise in basal and FSH stimulated inhibin secretion by the cryptorchid testis which seems to be due to a direct effect of temperature on Sertoli cells. The subsequent decline in inhibin secretion could be due to the disruption of the seminiferous epithelium. PMID- 2564338 TI - Comparison of the effects of glucagon-like peptide-1-(1-37) and -(7-37) and glucagon on islet hormone release from isolated perfused canine and rat pancreases. AB - Recently, it has been demonstrated that glucagon-like peptide-1 (GLP-1)-(7-37) possesses a potent insulinotropic activity. In this paper, we compared the effects of GLP-1-(1-37) and -(7-37) and glucagon on insulin, glucagon, and somatostatin release from isolated perfused canine and rat pancreases under the perfusate condition of 5.5 mM glucose plus arginine. With canine pancreas perfusion, 1 nM GLP-1-(7-37) was more potent in stimulating insulin and somatostatin release than was the same dose of glucagon [stimulation to 375 +/- 36% vs. 302 +/- 28% of the basal level for insulin (P less than 0.05); 724 +/- 129% vs. 311 +/- 33% of the basal level for somatostatin (P less than 0.01)]. GLP 1-(1-37) (1 nM) did not stimulate either insulin or somatostatin release. GLP-1 (7-37) (1 nM) decreased the glucagon level of the effluent perfusate to 67.2 +/- 3.4% of its basal level; but 1 nM GLP-1-(1-37) did not. Glucagon (1 nM) decreased GLP-1-like immunoreactivity to 64.0 +/- 5.2% of its basal level. With rat pancreatic perfusion, the minimal dose for stimulation of insulin release was 100 nM for GLP-1-(1-37), 0.1 nM for GLP-1-(7-37), and 1 nM for glucagon, respectively. Glucagon release was partially inhibited by 100 nM GLP-1-(1-37) and 1 and 10 nM GLP-1-(7-37). The present results indicate that 1) since GLP-1-(7-37) is released from the intestine, it might be an important incretin candidate along with gastric inhibitory peptide; and 2) the release of proglucagon-derived peptides from pancreatic A-cells is regulated by autofeedback through glucagon and GLP-1. PMID- 2564339 TI - Insulin-like growth factor-I action on growth hormone secretion and messenger ribonucleic acid levels: interaction with somatostatin. AB - Insulin-like growth factor I (IGF-I) suppresses GH gene transcription and GH secretion, while somatostatin (SRIF) only suppresses GH secretion. The interaction of these inhibitors of GH was, therefore, tested in primary rat pituitary cells grown in serum-free medium. Maximal inhibition of GH secretion (to 30% of the control value) was achieved by 13 nM IGF-I, while 5 nM SRIF suppressed GH to 36% of control secretion. The respective ED50 values for IGF-I and SRIF inhibition of GH secretion were similar (approximately 2.5 nM). Treatment of cells with the two agents together resulted in a further inhibition of basal GH secretion to 18% of control untreated cells (P less than 0.005). Increasing doses of SRIF (2.5-10 nM) in the presence of IGF-I caused a dose dependent suppression of GH secretion. PRL levels were not altered by these treatments, indicating a selective effect on GH. GRH-induced GH secretion was further attenuated by combined IGF-I and SRIF treatment compared to the effect of either of these two agents alone. Northern analysis showed that IGF-I suppressed GH mRNA transcripts, while SRIF did not alter GH mRNA levels. The results indicate that physiological concentrations of both IGF-I and SRIF suppress long term basal GH secretion. Only IGF-I alters GH mRNA levels. These two peptides, therefore, appear to attenuate in vitro GH secretion by different mechanisms. PMID- 2564341 TI - Mechanism of action of cysteinyl leukotrienes on glucose and lactate balance and on flow in perfused rat liver. Comparison with the effects of sympathetic nerve stimulation and noradrenaline. AB - Rat livers were perfused at constant pressure via the portal vein with media containing 5 mM glucose, 2 mM lactate and 0.2 mM pyruvate. 1. Leukotrienes C4 and D4 enhanced glucose and lactate output and reduced perfusion flow to the same extent and with essentially identical kinetics. They both caused half-maximal alterations (area under the curve) of carbohydrate metabolism at a concentration of about 1 nM and of flow at about 5 nM. The leukotriene-C4/D4 antagonist CGP 35949 B inhibited the metabolic and hemodynamic effects of 5 nM leukotrienes C4 and D4 with the same efficiency, causing 50% inhibition at about 0.1 microM. 2. Leukotriene C4 elicited the same metabolic and hemodynamic alterations with the same kinetics as leukotriene D4 in livers from rats pretreated with the gamma glutamyltransferase inhibitor, acivicin. 3. The calcium antagonist, nifedipine, at a concentration of 50 microM did not affect the metabolic and hemodynamic changes caused by 5 nM leukotriene D4. The smooth-muscle relaxant, nitroprussiate, at a concentration of 10 microM reduced flow changes, without significantly affecting the metabolic alterations. 4. Leukotriene D4 not only reduced flow; it also caused an intrahepatic redistribution of flow, restricting some areas from perfusion. Thus, leukotrienes increased glucose and lactate output directly in the accessible parenchyma and, in addition, indirectly by washout from restricted areas during their reopening upon termination of application. 5. The phospholipase A2 inhibitor, bromophenacyl bromide, but not the cyclooxygenase inhibitor, indomethacin, at a concentration of 20 microM reduced the metabolic and hemodynamic effects of 5 mM leukotriene D4. 6. Stimulation of the sympathetic hepatic nerves with 2-ms rectangular pulses at 20 Hz and infusion of 1 microM noradrenaline increased glucose and lactate output and decreased flow, similar to 10 nM leukotrienes C4 and D4. The kinetics of the metabolic and hemodynamic changes caused by the leukotrienes differed, however, from those due to nerve stimulation and noradrenaline. 7. The leukotriene-C4/D4 antagonist, CGP 35949 B, even at very high concentrations (20 microM) inhibited the metabolic and hemodynamic alterations caused by nerve stimulation or noradrenaline infusion only slightly and unspecifically.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2564340 TI - Regulation and patterns of endogenous and exogenous gene expression during differentiation of embryonal carcinoma cells. AB - Embryonal carcinoma (EC) cells offer an interesting model system for evaluating differentiation because the cells are pluripotent, thus resembling germ cells and embryonic stem cells, and because a number of agents have been defined that are capable of promoting the differentiation of these cells. This chapter examines how EC cells might be triggered to differentiate, with emphasis on retinoic acid because this compound is a potent, naturally occurring inducer that has been studied extensively in this system. The nature of alterations in gene expression during EC cell differentiation is reviewed from the perspective of evaluating whether these changes are likely to be responsible for, or a result of, the differentiation event. Finally, we consider in molecular terms why EC cells, but not their differentiated derivatives, are refractory to the expression of many viral genomes following infection. Based upon these studies, we propose that fundamental changes in gene expression that are observed when differentiation is triggered in EC cells are likely to be due to the disappearance or neutralization of strong repressor elements. PMID- 2564342 TI - Secretion-stimulating and secretion-inhibiting hormones stimulate high-affinity pertussis-toxin-sensitive GTPases in membranes of a pituitary cell line. AB - Different peptide hormones influence hormone secretion in pituitary cells by diverse second messenger systems. Recent data indicate that luteinizing-hormone releasing hormone (LHRH) stimulates and somatostatin inhibits voltage-dependent Ca2+ channels of GH3 cells via pertussis-toxin-sensitive mechanisms [Rosenthal et al. (1988) EMBO J. 7, 1627-1633]. In other pituitary cell lines, somatostatin has been shown to cause a pertussis-toxin-sensitive decrease in adenylate cyclase activity, and LHRH and thyrotropin-releasing hormone (TRH) stimulate phosphoinositol lipid hydrolysis in a pertussis-toxin-independent manner. Whether stimulation of Ca2+ influx by TRH is affected by pertussis toxin is not known. In order to elucidate which of the hormone receptors interact with pertussis-toxin sensitive and -insensitive G-proteins, we measured the effects of LHRH, somatostatin and TRH on high-affinity GTPases in membranes of GH3 cells. In control membranes, both LHRH and TRH stimulated the high-affinity GTPase by 20%, somatostatin by 25%. Maximal hormone effects were observed at a concentration of about 1 microM. Pretreatment of cells with pertussis toxin abolished pertussis toxin-catalyzed [32P]ADP-ribosylation of 39-40-kDa proteins in subsequently prepared membranes and reduced basal GTPase activity. The toxin also reduced by more than half the increases in GTPase activity induced by LHRH and TRH; stimulation of GTPase by somatostatin was completely suppressed. Stimulation of adenylate cyclase by vasoactive intestinal peptide (VIP) was not impaired by pretreatment of cells with pertussis toxin. Somatostatin but not LHRH and TRH decreased forskolin-stimulated adenylate cyclase activity. The results suggest that the activated receptors for LHRH and TRH act via pertussis-toxin-sensitive and -insensitive G-proteins, whereas effects of somatostatin are exclusively mediated by pertussis-toxin-sensitive G-proteins. PMID- 2564344 TI - Glutamate and aspartate immunoreactivity in cortico-cortical neurons of the sensorimotor cortex of rats. AB - Retrograde transport of tracers and immunocytochemistry have been used to determine if association and callosal neurons in the primary motor and somatosensory cortex of rats contain high levels of glutamate or aspartate and may, thus, use these amino acids as neurotransmitter. After tracer injections in these areas, about 65% of the retrogradely labeled neurons in layer V in the ipsilateral or contralateral hemisphere are immunopositive for glutamate. Lower percentages of double-labeled neurons are found in layers III, VI, and II. Similar results are obtained when sections are processed for aspartate immunoreactivity. About 90% of retrogradely labeled neurons are immunopositive in sections incubated with a mixture of both glutamate-and aspartate antisera. These results suggest that a large fraction of cortico-cortical neurons are immunoreactive for either one amino acid but not for both. It is proposed that neurons with high levels of one amino acid use this as neurotransmitter; high levels of glutamate and aspartate are likely to be present in a fraction of neurons which may release both amino acids or a substance closely related to these. PMID- 2564343 TI - Cholecystokinin and tyrosine hydroxylase messenger RNAs in neurons of rat mesencephalon: peptide/monoamine coexistence studies using in situ hybridization combined with immunocytochemistry. AB - The cellular localization of neurons expressing cholecystokinin (CCK) and tyrosine hydroxylase (TH) mRNAs was analysed in rat ventral mesencephalon using in situ hybridization techniques with both complementary DNA and synthetic oligonucleotide probes. Cell bodies distributed throughout the substantia nigra, ventral tegmental area, interfascicular nucleus, midline raphe nuclei, and central and ventral periaqueductal grey matter were found to contain CCK mRNA or TH mRNA as indicated by high densities of grains overlying the perikarya. The in situ hybridization technique was combined with immunocytochemistry on the same tissue section to localize the peptide or enzyme within its respective mRNA containing somata. In addition, the presence of TH immunoreactivity was demonstrated within cell bodies labeled for CCK mRNA and immunostaining for CCK was detected within TH mRNA-containing neurons. In the medial geniculate nucleus a strong labeling for CCKmRNA was observed, in spite of the fact that so far no CCK-like immunoreactivity has been demonstrated in perikarya in this nucleus. The specificity of the probes was verified by RNA blot hybridization. These results confirm recent double-labeling immunocytochemical studies and further characterize the coexistence of CCK and TH at the level of their mRNAs as well as their post-translational products in a large population of mesencephalic dopamine neurons known to project to forebrain areas. PMID- 2564345 TI - Induction of hemopoietic chimerism in the caprine fetus by intraperitoneal injection of fetal liver cells. AB - Intraperitoneal injection of allogeneic liver cells from 43-day-old male fetuses into normal 60-day female goat fetuses resulted in persistent hemopoietic chimerism in surviving recipients without clinical evidence of graft-versus-host disease. Transplantation of normal fetal liver cells into preimmunocompetent goat fetuses affected with beta-D-mannosidosis may provide an alternative strategy for evaluating hemopoietic stem cell transplantation in the treatment of human lysosomal storage diseases. PMID- 2564346 TI - Apoptotic hepatocytes become insoluble in detergents and chaotropic agents as a result of transglutaminase action. AB - Physiological deletion of cells ensues programmed death which involves formation of apoptotic bodies with fragmented DNA. Here we report that apoptotic hepatocytes are insoluble in detergents, urea, guanidine hydrochloride, reducing agents and thereby can be isolated from rat liver following collagenase treatment. They are wrinkled, spherical structures similar to cornified envelopes of epidermis by phase-contrast microscopy and show irregular, globular morphology by scanning-electron microscopy. Part of their DNA content is cleaved into nucleosomal and oligonucleosomal fragments. Their insolubility, like that of the cornified envelope, is evoked by epsilon-(gamma-glutamyl)lysine and N1,N8 bis(gamma-glutamyl)spermidine protein cross-linking bonds formed by transglutaminase. PMID- 2564347 TI - Particular processing of pro-opiomelanocortin in Xenopus laevis intermediate pituitary. Sequencing of alpha- and beta-melanocyte-stimulating hormones. AB - alpha- and beta-melanocyte-stimulating hormones (alpha-MSH and beta-MSH) have been isolated from Xenopus laevis neurointermediate pituitary and microsequenced. Intracellular alpha-MSH is not N-acetylated after proteolytic processing of pro opiomelanocortin in contrast to mammalian alpha-MSHs. There is a high preservation of the melanotropic amino acid sequence common to all MSHs although in Xenopus beta-MSH a histidine residue replaces the glutamic acid residue found in position 8 of mammalian beta-MSHs. PMID- 2564348 TI - [The effect of stimulation frequency on the speed of isometric myocardial relaxation in mammals]. AB - The relaxation of tension of the rabbit and rat heart right ventricle papillary muscles was estimated by the characteristic time t30. An increase in the stimulation rate as well as an increase in extracellular concentration of Ca ions decelerated the relaxation in the rabbit myocardium, whereas in the rat myocardium a shortening of interimpulse interval accelerated isometric relaxation. Constant administration of reserpine and beta-blocking agents did not alter the effects. The mechanism of the stimulation rate effect on the velocity of relaxation are discussed. PMID- 2564349 TI - The incidence of pregnancy induced hypertension in southeast Turkey. AB - One hundred twenty patients with pregnancy-induced hypertension (PIH) were delivered during 1986. The incidence of pre-eclampsia and eclampsia, per 1000 births, was 103 and 19, respectively. Two-thirds of eclamptics were nulliparous and three-quarters were less than 25 years of age. All patients with eclampsia had convulsions before admission. Although maternal mortality was 4% in patients with pre-eclampsia and 0% in those with eclampsia, the fetal mortality rate was 32% in pre-eclamptic and 60% in eclamptic patients. PMID- 2564350 TI - Accuracy of prediction of gestational age by ultrasound measurement of biparietal diameter in Nigerian women. AB - The accuracy of assessment of gestational age by ultrasound measurement of biparietal diameter (BPD) was evaluated in 84 Nigerian women. Ultrasound and menstrual delivery dates were compared in these women who had no complications of pregnancy and delivered infants whose birthweights were appropriate for 40 weeks. The results showed that ultrasound dating was more accurate than menstrual dating as evident from the number of women who delivered on and within 1 or 2 weeks of predicted delivery dates. It is concluded that (1) our locally produced BPD normogram is as accurate as those from elsewhere and (2) the normogram is recommended for use in Nigerian women for accurate pregnancy dating. PMID- 2564351 TI - Cervical ripening with intravaginal prostaglandin E2 gel. AB - Fifty-four patients were randomized in a double blind fashion into either a placebo group or a group treated with a 3 mg dose of an intravaginal prostaglandin E2 gel. The group receiving the prostaglandin E2 gel had a mean change in cervical score of 2.7 while the patients in the placebo group had no significant change. Even though a significantly higher number of patients in the prostaglandin group went into spontaneous labor, the incidence of cesarean section was not different between the two groups. Upon comparing the conflicting reports of various studies on the subject, it is concluded that the method of product preparation, particularly the source of prostaglandin E2 utilized and the choice of drug vehicle, may be important variables in determining treatment success, as measured in terms of decreased cesarean section rate. PMID- 2564352 TI - The development and evaluation of a computerised antenatal questionnaire. AB - A computerised questionnaire, for convenience entitled "PAM" (Programmed Aid for use in Midwifery), was created for use at the time of first antenatal assessment. The system required simple direct input by the expectant mother; and in response to a short descriptive letter, 73.4% of 349 women were prepared to try the new method. The response of 100 consecutive women who tried the system was further analysed. Seventy women liked PAM, four disliked the technique and the remainder had no strong feelings. Asked by the midwife which they would prefer, 46 women expressed a preference for the computer questionnaire and only nine would rather have had a traditional interview with a midwife asking the same questions. PAM or similar systems provide a new opportunity, both for research and as a means of reliably decentralizing the initial assessment of a pregnancy. PMID- 2564353 TI - The outcome of pregnancies complicated by preterm rupture of the membranes with and without cerclage. AB - Thirty-two patients with cerclage were compared to 76 patients without cerclage. All pregnancies were complicated with preterm (less than 36 weeks) rupture of the membranes (PTROM). The management following removal of the cerclage was the same for the two groups and consisted of conservative treatment unless chorioamnionitis ensued. It was found that under similar conditions and treatment there was no significant difference between the outcomes of PTROM-complicated gestations with and without a cerclage. The low added risk of cerclage to PTROM should not deter its use when indicated. PMID- 2564354 TI - The relation of the true conjugate to maternal height and obstetric performance in Ghanaians. AB - The true conjugate was determined intraoperatively with a caliper in 114 Ghanaian women and was correlated with their height, obstetric performance and fetal dimensions. Those patients undergoing cesarean section for cephalopelvic disproportion (Group Ia) were found to have a significantly shorter mean true conjugate (9.54 cm +/- 0.63 S.D.) and mean body height (152.68 cm +/- 5.46 S.D.) and a smaller true conjugate - fetal biparietal diameter difference (10.93 mm) than those who had no cephalopelvic disproportion (Group Ib) and whose mean measurements were 10.61 cm +/- 0.81 S.D., 157.20 cm +/- 5.69 S.D. and 21.50 mm, respectively (P = 0.0001). Recommendations for appropriate referral of rural clinic patients and for selection of patients for repeat cesarean sections are based on the above findings. PMID- 2564355 TI - Fetal macrosomia and pregnancy outcome in Lagos. AB - In an analysis of 6376 singleton births the prevalence of macrosomia was 4.9%; the attending perinatal mortality was 58/1000 compared to 18/1000 in controls. Eighty-three percent perinatal deaths occurred in unbooked patients after prolonged and neglected labor. Mortality and morbidity were weight related; the macrosomic baby delivered by section or by diabetic mother or that died was significantly heavier. There was strong association between maternal age, parity, diabetes, mild hypertension, previous history of a big baby and macrosomia in this study. Pregnancy was significantly prolonged with higher incidence of emergency sections and primary postpartum hemorrhage in mothers of macrosomic babies. Fetal sex does not appear to be an important factor in macrosomia. PMID- 2564356 TI - The effect of postplacental insertion of the spiked and the standard Lippes loop on uterine involution. AB - The spiked Lippes loop was inserted in 53 women and the standard loop in 55 women immediately following the delivery of the placenta after normal vaginal delivery. A group of 50 recently delivered women with no IUD was studied as a control. The cases were followed up for 1 year. Ultrasonic scanning was performed during the 6th week after insertion for 47 cases with the spiked loop and 43 cases with the standard loop. All loops were correctly positioned within the uterine cavity except in two cases; one using the spiked and the other using the standard loop. The loops were found displaced downwards and as there was excessive bleeding, the devices were removed. Regarding the effect of the IUD on uterine involution, it was found that the fundus to internal os length for the cases with IUDs was shorter than that among the controls, also this length among the standard loop users was significantly shorter than that of the controls (P less than 0.01). Postplacental IUD insertion did not delay uterine involution, on the contrary, it helped involution. Regarding the effect of lactation, the fundus to internal os length for the fully lactating women either using a loop or not, was found to be significantly shorter than that of the non-lactating cases (P less than 0.002). PMID- 2564357 TI - A 3-year evaluation of the TCu380Ag and the Cu-7. AB - The TCu380Ag and the Cu-7 intrauterine devices (IUDs) were evaluated in a randomized comparative clinical trial. A total of 198 women who had not recently been pregnant entered the trial in Manila, Philippines and were followed-up through 3 years postinsertion. The 36-month follow-up rates were impressively high at 80.3 for TCu 380A users and 76.7 for users of the Cu-7. Although the two IUDs demonstrated statistically similar termination rates, the findings suggest a higher efficacy for the TCu380Ag. Gross cumulative life-table pregnancy rates were 0.0 and 4.5 for the TCu380Ag and Cu-7, respectively, at 3 years. Overall 3 year continuation rates were 74.3 and 64.9 for users of the respective devices. PMID- 2564358 TI - Pregnancy in a non-communication rudimentary uterine horn. AB - A case of pregnancy in a rudimentary uterine horn is presented. The pregnancy ended in fetal demise at the end of the second trimester. The diagnosis was missed by ultrasonography on two occasions and was made only at laparotomy following failure to initiate uterine contractions. Histologic examination of the myometrium of the rudimentary horn revealed extensive interstitial fibrosis. A high index of suspicion of pregnancy in a rudimentary horn is recommended whenever adequate ecbolic infusions fail to induce myometrial contractions. PMID- 2564359 TI - Pregnancy and primary biliary cirrhosis. AB - Pregnancy is rarely encountered in patients with primary biliary cirrhosis. Ten pregnancies in nine patients have been reported in the literature. All five pregnancies that continued beyond the 31st week had increasing jaundice during late pregnancy. We describe a case of primary biliary cirrhosis with symptomatic onset at age 19, pregnancy a year later, with subsequent resolution of jaundice and pruritus. This course is different than most described by others. PMID- 2564360 TI - Vaginal sonographic visualisation of a cervical carcinoma. AB - In the present case, the vaginal sonographic picture of a cervical (collum) carcinoma, which was clinically non-apparent, but was histologically established through a fractionated curettage, led to a change in the surgical procedure in the sense that a primary operation according to Wertheim--Meigs instead of a knife conisation was performed. The postoperative histology showed a good correlation between sonographic tumor boundaries and the actual expansion of the carcinoma. PMID- 2564361 TI - Successful hemisterilization in a patient with uterus didelphus. AB - Women with uterine malformations frequently present with reproductive impairment. This case addresses the opposite problem, a woman with uterus didelphus who conceived with ease. Following three consecutive cesarean sections in the left uterine horn, she was offered sterilization for medical reasons, but refused on religious grounds. As a compromise, she accepted unilateral tubal closure in order to force future pregnancies to the right uterine horn. She has now had three pregnancies with two cesarean sections on that side. PMID- 2564362 TI - Embolization therapy in patients with severe arterial bleeding after hysterectomy. AB - The case history of a patient aged 41 years, who suffered recurrent hemorrhages following vaginal hysterectomy is reported. After several surgical interventions complete hemostasis was finally achieved by selective embolization of the internal iliacal artery. The application of this procedure is recommended in case of recurrent bleeding complications caused by pelvic surgery or neoplasms. PMID- 2564363 TI - New technique for managing second trimester intrauterine fetal death. AB - In this study, a new method for terminating second trimester pregnancies complicated by intrauterine fetal death is analysed. The technique consisted of a combination of extraamniotic ethacridine lactate with intramuscular sulprostone (16-phenoxy-omega-17,18,19,20 tetranor PGE2 methyl sulfonylamide). Objective documentation of the efficacy of this method was obtained by continuous monitoring of intrauterine pressure in two patients. The method was found to be simple, safe, cheap and effective and deserves increased acceptance. PMID- 2564364 TI - A requirement for protein phosphorylation in regulating the meiotic and mitotic cell cycles in echinoderms. AB - Populations of hormone-stimulated starfish oocytes and fertilized sea urchin eggs undergo synchronous meiotic and mitotic divisions. We have studied the requirement for protein phosphorylation during these events by testing the effects of 6-dimethylaminopurine (6-DMAP) upon the incorporation of [32P]orthophosphate. It was found that 6-DMAP blocked meiosis reinitiation and early cleavage and simultaneously inhibited protein phosphorylation, without changing the rate of [35S]methionine incorporation or pattern of protein synthesis. The protein, cyclin (54 kDa in starfish and 57 kDa in sea urchin), continues to be synthesized in the presence of 6-DMAP. This protein is destroyed at first and second cell cycles when 6-DMAP is added 30 min following fertilization but not when this drug is present before fertilization. Thus, cyclin breakdown does not depend on the completion of the nuclear events of M phase, and its time of breakdown is set at an early step between fertilization and first cleavage. Using tubulin immunostaining, we found that 6-DMAP did not affect the cortical microtubules and resting female centrioles of prophase arrested starfish oocytes, whereas it induced a precocious disappearance of spindle fibers when applied to hormone-stimulated oocytes. While an early addition of 6-DMAP precluded nuclear breakdown and spindle formation in both systems, a late treatment always allowed chromosome separation and centriole separation. Under these conditions pericentriolar tubulin persisted and could organize new spindles after the inhibitor was removed. It is suggested that (1) the assembly of cortical and centriolar-associated microtubules is not controlled by the same factors as spindle-associated tubulin; (2) specific proteins which are required for the cell to enter the following M-phase can become operative only via a process depending upon protein phosphorylation; (3) microtubule associated kinases may play an important role in MPF function and spindle dynamics. PMID- 2564365 TI - Increased basal glucose production and utilization in children with recent obesity versus adults with long-term obesity. AB - To characterize the abnormalities of glucose homeostasis and insulin action early in the course of human obesity, we studied in vivo glucose kinetics in seven children who were recently massively overweight. At time of study they were gaining weight at a rate of 13.5 +/- 1.4 kg/yr. They were compared with six age matched control subjects. Six adults with long-term obesity and five normal adults were studied in parallel. The obese children and adults were normoglycemic and hyperinsulinemic. We found that glucose production and utilization were remarkably higher in obese children (295 +/- 18 mg/min; 7.6 mg.kg-1 lean body mass.min-1) than in control children (129 +/- 13 mg/min; 4.4 mg.kg-1 lean body mass.min-1, P less than .01) and obese adults (151 +/- 8 mg/min; 3.1 +/- 0.3 mg.kg-1 lean body mass.min-1, P less than .01). Obese adults had normal rates of glucose production and utilization. Insulin- and non-insulin-mediated glucose uptake, estimated with somatostatin-induced suppression of endogenous insulin secretion, contributed almost equally to the excess glucose utilization observed in the obese children. When studied with the euglycemic-hyperinsulinemic clamp, obese children could not increase glucose disposal to the same extent as normal children and were not able to adequately suppress their endogenous glucose production. Recently obese children are therefore characterized by an increased basal glucose turnover rate and an already established insulin resistance of the liver and probably the skeletal muscles. PMID- 2564366 TI - A role for dopamine as an endogenous protective factor in the rat stomach. AB - Application of an irritant to the surface of the gastric mucosa confers protection against subsequent application of a damaging agent ("adaptive cytoprotection"). The possibility that this adaptive response is mediated via the release of nonprostaglandin mediators was examined using an ex vivo gastric chamber model, in which the irritant (1 M sodium chloride) could be applied to only one-half of the exposed mucosa. Rats were pretreated with indomethacin and one of various receptor antagonists, including cimetidine, pyrilamine, atropine, propranolol, phentolamine, cyproheptadine, haloperidol, and BN52021. In control studies, application of hypertonic saline to the left side of the mucosa reduced hemorrhagic damage induced by subsequent application of 70% ethanol by 71% +/- 4% (p less than 0.01). Of the receptor antagonists tested, only cyproheptadine and haloperidol significantly attenuated the degree of protection afforded by exposure to hypertonic saline. Subsequent dose-response studies with other serotonergic and dopaminergic antagonists suggested that dopaminergic receptors are involved in the adaptive response to the irritant. Topical application of a dopamine agonist, bromocriptine, or L-beta-3,4-dihydroxyphenylalanine, significantly reduced the extent of damage induced by subsequent application of 70% ethanol. The results of the gastric chamber studies were confirmed in conscious rats in which the irritant, dopamine agonist, and ethanol were administered orally. These results therefore suggest a role for endogenous dopamine as a mediator of the adaptive cytoprotection phenomenon. PMID- 2564367 TI - Relationship between serum growth hormone levels and the brain and pituitary content of immunoreactive somatostatin in the goldfish, Carassius auratus L. AB - In this study, the relationships between endogenous brain and pituitary immunoreactive somatostatin (irSRIF) and circulating growth hormone (GH) levels in the goldfish were examined using two approaches. First, the amount of irSRIF in extracts of the pituitary gland and various brain regions was measured by radioimmunoassay several times throughout the year and was compared to serum GH levels at each time. The amounts of irSRIF in extracts of the pituitary gland, hypothalamus, and telencephalon were found to be inversely related to seasonal changes in serum GH levels, such that irSRIF was highest in these regions when serum GH levels were lowest (November and February). Conversely, irSRIF in these regions was lower in May, June, and July when serum GH levels were highest. These results suggest that endogenous irSRIF in the pituitary and forebrain may participate in the regulation of seasonal changes in serum GH levels in the goldfish. In extracts from other brain regions (thalamus + midbrain and cerebellum + medulla), some changes in the amount of irSRIF were observed among the various sample times, but these variations were not related to changes in serum GH levels. In a second set of experiments, the origin of irSRIF fibers innervating the goldfish pituitary gland was examined by using brain lesioning techniques to destroy regions of the forebrain known to contain irSRIF perikarya and fibers, and subsequently measuring the amount of irSRIF in the pituitary gland. Lesions in the preoptic area of the forebrain resulted in increased serum GH levels concomitant with a decrease in pituitary irSRIF content. This provides direct evidence that the preoptic area is the origin of a somatostatinergic projection inhibiting GH secretion from the goldfish pituitary. Lesions centered in the nucleus lateral tuberis (NLT) pars anterioris did not influence serum GH levels or the pituitary content of irSRIF. In contrast, more posterior lesions centered in the NLT pars posterioris (NLTp) resulted in a dramatic reduction in the amount of irSRIF in the pituitary. This suggests that the majority of irSRIF projections to the goldfish pituitary pass through the area destroyed by the lesion centered in the NLTp; it is also possible that perikarya within this area may be the origin of at least some of the irSRIF-containing fibers in the goldfish pituitary. Together, results from the present study provide evidence of a functional relationship between circulating levels of GH and endogenous brain and pituitary irSRIF in the goldfish. PMID- 2564368 TI - Hepatic encephalopathy in thioacetamide-induced acute liver failure in rats: characterization of an improved model and study of amino acid-ergic neurotransmission. AB - An imbalance of excitatory and inhibitory amino acid-ergic neurotransmission has been suggested to play a role in the pathogenesis of hepatic encephalopathy. For further evaluation of this hypothesis, several parameters of amino acid-ergic neurotransmission were studied in rats with acute liver failure induced by the administration of 300 mg per kg thioacetamide by gavage on two consecutive days. By appropriate supportive care, hypoglycemia, renal failure and hypothermia were avoided. Rats were monitored clinically and neurologically. Hepatic encephalopathy evolved in four distinct, easily recognizable stages. Light and electron microscopic examination of brains of rats with hepatic encephalopathy revealed only a slight swelling of nuclei of neurons and astrocytes without signs of neuronal degeneration or brain edema. In rats with hepatic encephalopathy, the concentrations of GABA, glutamate and taurine were decreased in the cerebral cortex, the hippocampus and the striatum, whereas those of aspartate and glycine were unchanged or increased. GABAA and benzodiazepine receptors were studied as parameters for the postsynaptic GABAA-benzodiazepine receptor complex, glutamic acid decarboxylase as parameter for presynaptic GABA-ergic neurons and stimulation of benzodiazepine binding by GABA as a parameter for a GABA-mediated postsynaptic event. None of these parameters was different in hepatic encephalopathy as compared to controls. Similarly, Ca++/Cl(-)-dependent and independent glutamate receptors as parameters for glutamatergic neurons were unchanged in rats with hepatic encephalopathy. Thus, in rats with thioacetamide induced liver failure and hepatic encephalopathy, changes of the concentrations of neurotransmitter amino acids occur in the brain. Other neurochemical parameters, however, failed to identify alterations of GABA-ergic or glutamatergic neurotransmission in hepatic encephalopathy. PMID- 2564369 TI - Studies on CD11a and CD18 molecules with two new monoclonal antibodies: differential myelomonocytic antigen expression of PMA treated HL60 and U937 cell lines. AB - Biochemical and functional aspects as well as features of cellular distribution of the differentiation groups CD11a and CD18 were studied in the course of a detailed characterization of two new monoclonal antibodies which recognize the alpha (GRS3) and beta (GRF1) chains of the LFA-1 antigen. Both MAbs inhibited homotypic aggregation of an EBV cell line. In contrast, only GRF1 (anti-beta chain) was able to inhibit granulocyte aggregation as well. Different myeloid monocyte antigen modulation was noted in PMA induced macrophage differentiation of the U937 and HL60 cell lines. PMA treated HL60 cells showed increased expression of alpha M (CD11b) and alpha X (CD11c) antigens but no change in HLA DR or CD14 antigen expression. No variation in the expression of LFA complex antigen (CD11a, b and c, or CD18) was observed on U937 cells, which on the other hand presented de novo expression of HLA-DR and CD14 antigens. PMID- 2564370 TI - Exon duplication and triplication in the human T-cell receptor gamma constant region genes and RFLP in French, Lebanese, Tunisian, and black African populations. AB - The human T-cell receptor gamma (TCRG) locus comprises 14 variable genes (TRGV), five joining segments (TRGJ), and two constant region genes (TRGC). The constant gamma 1 gene, TRGC1, consists of three exons, whereas the TRGC2 gene contains four or five exons due to the duplication or triplication of exon 2 and spans 9.5 kb or 12 kb, respectively. In this paper, we define the alleles of the T-cell receptor gamma J2 and C2 genes, and we show that two Hind III allelic fragments, 5.4 kb and 8 kb, characterize unambiguously the C2 gene with duplication or triplication of exon 2. We show also that the cDNA of the HPB-MLT cell line results from the transcription of an allelic TRGC2 gene with duplicated exon 2. We propose a model involving unequal crossing-overs to explain the organization and the evolution of the TRGC locus. Moreover, we analyze the TCRG haplotypes in four different populations (French, Lebanese, Tunisian, and Black African) to underline their interest for population genetics. PMID- 2564371 TI - Restriction fragment length polymorphisms of the mouse T-cell receptor gene families. AB - We have studied the restriction fragment length polymorphisms (RFLPs) found in the germline T-cell receptor genes of 25 inbred Mus musculus strains and 8 wild Mus species. Included in the inbred mice tested were several strains which spontaneously develop systemic autoimmune disease. Extensive polymorphism was evident for the variable (V) gene segments of the alpha gene family for both the inbred strains and wild mouse species. Changes in the total number of bands hybridizing with probes for V alpha gene segments suggest that members of a V alpha gene segment subfamily are not closely linked, but are interspersed with members of other subfamilies; that expansion and contraction of the multimembered subfamilies may be an important diversifying factor. Our data obtained with beta gene probes revealed genomic diversity that is much more limited than that seen for the alpha locus. Analysis of inbred mice with probes for the gamma gene locus revealed some RFLPs, but little evidence of expansion or contraction in the numbers of gene segments. Among the autoimmune mice, NZW, NZB, and BXSB/MpJ all display distinctive differences with alpha gene probes. NZW mice have a large deletion of the beta gene family, which has been reported previously. We found no differences to distinguish the MRL/MpJ lpr/lpr mice from non-autoimmune strains. PMID- 2564372 TI - Recombinational sites observed in naturally occurring intra-RT1 recombinant strains. PMID- 2564373 TI - Role of afferent renal nerves in spontaneous hypertension in rats. AB - In the present study we examined sympathetic function and baroreceptor reflex sensitivity in adult spontaneously hypertensive rats (SHR) after a selective transection of afferent renal nerves in the prehypertensive and established phases of hypertension. Renal deafferentation performed between 3 and 4 weeks after birth did not influence the course of the development of high blood pressure when compared with sham-operated rats. Mean arterial pressure, heart rate, and plasma norepinephrine concentrations were similar in both groups when measured at 13 weeks after renal deafferentation. However, blood pressure responses to ganglionic blockade with hexamethonium were significantly reduced in the renal deafferented SHR. Baroreceptor reflex sensitivity, assessed by heart rate responses to blood pressure changes induced by phenylephrine and nitroprusside, was significantly enhanced in these rats. When renal deafferentation was performed in adult SHR with established hypertension, mean arterial pressure decreased slightly but significantly by 5%. Heart rate, plasma norepinephrine concentrations, and responses to hexamethonium were not affected by this procedure. However, in the renal deafferented adult SHR, heart rate responses to phenylephrine but not to nitroprusside were significantly increased. Thus, in contrast to efferent renal nerves, afferent renal nerves do not play an important role in the development and maintenance of high blood pressure in SHR, but may contribute to the mechanisms that alter sympathetic function and baroreceptor reflex sensitivity in SHR during the development of hypertension. PMID- 2564374 TI - Molecular cloning and nucleotide sequence of the colonization factor antigen I gene of Escherichia coli. AB - The colonization factor antigen I (CFA/I) gene has been isolated and sequenced. The amino acid sequence of CFA/I deduced from the nucleotide sequence is composed of 170 amino acids. The first 23 amino acids are considered to be the signal peptide of the CFA/I protein since they are not present in the protein sequence. Among the remaining amino acids, only two are different from the protein sequence: amino acid position 76 is an aspartic acid instead of an asparagine, and position 97 is a serine instead of an alanine. The CFA/I gene has a typical Shine-Dalgarno sequence located 10 base pairs (bp) upstream from the initiation codon. The sequence TACAAT located 48 bp upstream from the initiation codon was tentatively designated the -10 sequence of the CFA/I gene promoter. No sequences homologous to the consensus -35 promoter sequence was found. A pair of inverted repeat sequences followed by a stretch of eight A's are located 45 bp downstream from the termination codon of the CFA/I gene; this region may be a rho independent transcriptional terminator. PMID- 2564375 TI - Failure of pertussis toxin to inhibit activation of guanylate cyclase by the heat stable enterotoxin of Escherichia coli (STa) in the T84 cell line. AB - The heat-stable enterotoxin (STa) of Escherichia coli causes intestinal secretion by stimulating guanylate cyclase, an enzyme believed to be distinct from the STa receptor. Pertussis toxin (PT) has been reported to block the ability of STa to stimulate guanylate cyclase in rat intestinal mucosa (S. A. Epstein, R. A. Giannella, and H. J. Brandwein, FEBS Lett. 203:44-48, 1986). This suggested that a guanine nucleotide regulatory protein (G protein) coupled the STa receptor to guanylate cyclase, a function not previously recognized for G proteins. We sought to explore this phenomenon and, if possible, to identify this G protein. Initial experiments with the human colon carcinoma cell line T84 revealed that higher than-expected concentrations (1 micrograms/ml) of PT were needed to intoxicate cells, as assessed by ADP-ribosylation of endogenous PT substrate, but that 99 to 100% intoxication could be achieved. Homogenates made from fully intoxicated cells did not differ from controls in basal or STa-stimulated guanylate cyclase activity, and cyclic GMP accumulation in intact T84 cells was not changed by PT treatment. We conclude that a PT-sensitive G protein is not involved in the stimulation of cyclic GMP production by the enterotoxin STa. PMID- 2564376 TI - Actinomyces viscosus fibril antigens detected by immunogold electron microscopy. AB - Strains representing taxonomic clusters of Actinomyces viscosus and Actinomyces naeslundii were studied by indirect immunogold electron microscopy with either monospecific anti-type 1 and anti-type 2 rabbit antibodies or species-specific monoclonal antibodies. The monoclonal and anti-type 2 antibodies localized on long fibrils, whereas the anti-type 1 antibodies mostly localized close to the cell body or on shorter appendages. PMID- 2564377 TI - Correlative study on neuro-endocrine differentiation and presence of somatostatin receptors in breast carcinomas. AB - To correlate the expression of neuro-endocrine (NE) markers with the presence of somatostatin receptors (SSR) in human breast cancer, a series of 100 cases was stained with the Grimelius silver procedure, by immunocytochemistry with specific NE markers (i.e., chromogranin A and B and synaptophysin) and by autoradiography for SSR. Nine cases were positive for at least one NE marker, while 4 cases were positive for all of them. SSR were detected in 17 cases, either with low (10 cases) or with high receptor density (7 cases). A highly significant correlation was established between the expression of NE markers and high SSR density. Our findings are additional evidence of the existence of a group of breast cancers showing morphological and cytochemical similarity with neuro-endocrine tumors present in other organs. The identification of this type of breast cancer is of biological and potential therapeutic interest. PMID- 2564378 TI - Restriction fragment length polymorphism of the L-myc gene and susceptibility to metastasis in renal cancer patients. AB - We examined Southern blot analyses of normal and tumor DNAs from 50 patients with sporadic renal cancer, using the human L-myc oncogene fragment as a hybridization probe. Our purpose was to study the relationship between the restriction fragment length polymorphism (RFLP) of the L-myc and the frequencies of metastases. There was no individual difference in patterns of L-myc RFLP between normal and tumor tissue DNAs digested with EcoRI. The patients were classified into 3 genetic types according to the polymorphic patterns defined by the 2 alleles [10-kilobase (kb) and 6.6-kb fragments]. The relative ratios of the 3 genotypes in the renal cancer patients were similar to those seen in healthy Japanese. However, of 16 patients who exhibited distant organ metastases at the time of surgery, only one was a 10-kb fragment homozygote. The incidence of distant metastases in 10-kb homozygotes was significantly lower than that in 6.6-kb homozygotes plus heterozygotes (p = 0.06). These results basically correspond to the previous findings in the lung cancer patients, and suggest that L-myc RFLP is a widely applicable genetic marker to predict prognosis in cancer patients. PMID- 2564379 TI - T-lymphocyte subsets in patients with idiopathic dilated cardiomyopathy. AB - T-cell subsets were measured in the peripheral blood of 33 patients with heart failure from idiopathic dilated cardiomyopathy, 22 patients with heart failure from other causes, and 33 normal controls. Mean T-suppressor cell percentage was 30% in normals, 21% in patients with idiopathic dilated cardiomyopathy whose duration of symptoms was less than 1 year (P = 0.0005), and 26% in those with symptoms for greater than 1 year (P = 0.05). Similarly, percentage of T suppressor cells in the group with heart failure from causes other than idiopathic dilated cardiomyopathy was significantly lower (23%; P = 0.005) in those with short duration of symptoms. When both heart failure groups were combined those with symptoms for less than 1 year had significantly lower T suppressor frequencies (22%) than those with symptoms for more than 1 year (P = 0.015). Multivariate analysis identified duration of symptoms and age as the only independent predictors of T-suppressor cell frequencies. Decreased percentage of T-suppressor cells in patients with idiopathic dilated cardiomyopathy may be an epiphenomenon related to duration of heart failure. This should be taken into account in assigning an etiologic mechanism for T-suppressor cells in idiopathic dilated cardiomyopathy. PMID- 2564380 TI - Physician assistant supervisor regulations. PMID- 2564381 TI - Subdural empyema complicating frontal and ethmoid sinusitis. PMID- 2564382 TI - Antigenic expression of aminopeptidase M, dipeptidyl-peptidase IV and endopeptidase by primary cultures from rabbit kidney proximal tubule. AB - Techniques using microdissected tubules from rabbit kidney allow the isolation of well defined segments which can be cultured to obtain pure renal cell epithelia. From microdissected proximal tubules, we obtained epithelia the cells of which exhibit some of the antigenic expressions of the initial proximal cells. For this purpose, we used three monoclonal antibodies raised against apical brush border membranes of the proximal tubules. We determined with precision the identity and some of the molecular characteristics of the antigens bound by these three antibodies and found that they correspond to three hydrolases present in the brush borders of proximal renal cells (amino-peptidase, dipeptidyl-peptidase IV and endopeptidase). These apical markers are expressed by the growing cells of primary cultures from proximal tubules, suggesting strongly that they are effectively proximal cells and that no appreciable dedifferentiation occurred during the growth process. We have also shown that apical expression of these hydrolases on the plasma membrane of the epithelium occurred only after several days of culture and determined the complete polarization of the cells. Electron microscopy studies confirmed the degree of polarization of the cultured cells by the presence of numerous microvilli on their apical face. PMID- 2564383 TI - An increased risk of insulin-dependent diabetes mellitus (IDDM) among HLA DR4,DQw8/DRw8,DQw4 heterozygotes. AB - Serological HLA typing of 92 insulin-dependent diabetes mellitus (IDDM) patients and 300 healthy controls was performed by the immunomagnetic typing technique. We found an increased risk of IDDM among DR4/w8 heterozygotes, similar to that seen for DR3/4 heterozygotes. The DQ alleles of these DR4/w8 patients were therefore established. When hybridized with a cDNA DQB probe, BamHI-digested DNA from eight out of the nine DR4/w8 patients revealed only one single 10.8-kb DQB1-specific fragment typical of DQw4 and DQw8. DNA from all DR4/w8 patients also hybridized to an oligonucleotide probe corresponding to amino acids (aa) 23-30 of the beta chain of DQw8. However, using the oligonucleotide probe, the staining intensity was found to be only half of that seen when DNA from DQw8 homozygotes was used instead, suggesting that the eight DR4/w8 patients had DQw8 in a single dose and carried the DQw4 allele at the DRw8 haplotype. Therefore, eight of nine DR4/w8 IDDM patients seemed to be DR4,DQw8/DRw8,DQw4, which, thus, may be associated with susceptibility to develop IDDM. One common explanation of IDDM susceptibility for DR4,DQw8/DRw8,DQw4 and DR3,DQw2/DR4,DQw8 heterozygotes may be that they share similar DQ alpha beta epitopes encoded by DQA and DQB genes in trans. PMID- 2564384 TI - Beginnings and main directions of psychoneuroimmunology. AB - The present paper offers a survey of advances in the domain of psychoneuroimmunology. Major topics covered are the role of the nervous system in regulation of the immune system functions, neuromediator and neuroendocrine mechanisms for regulation of the lymphoid cell activity and hypothalamic involvement in modulation of the activity of bone marrow as a source of stem cells. Emphasis has been on the neurophysiological correlates of the organism's reaction to antigen. Attention has been given to the work of A.D. Speransky and discussion of the psychoneuroimmunologic research based on literature data and findings from our own studies is presented in the historical context. PMID- 2564385 TI - Pediatricians' use of chaperones when performing gynecologic examinations on adolescent females. AB - This study assesses the influence of the interest and skill in adolescent health care on a pediatrician's use of a chaperone during the pelvic examination. A national sample of 558 pediatricians selected at random and 384 members of the American Academy of Pediatrics' Section on Adolescent Health (SAH) completed questionnaires assessing their perceived skills in and practice of adolescent medicine. Some 90.4% of the non-SAH males and 31.0% non-SAH females always used a chaperone compared to 69.4% of SAH males and 12.5% of SAH females (p less than or equal to 0.05). The actual frequency of chaperone use was less among SAH members than non-SAH pediatricians (p less than or equal to 0.013) and less among females in both groups (p less than or equal to 0.0001). Among male physicians, the variables found to have the strongest association with a less frequent use of chaperones included the frequency that oral contraceptives were prescribed, perceived skill in providing contraceptive counseling, percent of adolescents in the pediatrician's practice, and the frequency with which confidential services were provided. These correlations were stronger among SAH members than non-SAH pediatricians. Among female SAH members, not using a chaperone was associated with a higher perceived skill in managing sexually transmitted diseases, the frequency with which they performed pelvic examinations, and the percent of adolescents in their practice. Among non-SAH female physicians, the perceived skill to perform a pelvic examination and provide contraceptive counseling were associated with increased chaperone use. These data suggest that many factors other than gender and medicolegal influences are associated with the physician's decision concerning the use of a chaperone. PMID- 2564386 TI - Dirofilariasis. PMID- 2564387 TI - Evaluation of cimaterol for finishing swine including a drug withdrawal period. AB - One hundred fifty crossbred pigs (55 kg) were allotted by weight, sex and litter to a randomized complete-block design with five dietary treatments, six blocks per treatment and five pigs per pen with sex equalized across treatments. Corn soybean meal-based diets (.65% lysine) with 0, .25 and .5 mg/kg cimaterol were fed, on an ad libitum basis, to pigs slaughtered at an average pen weight of 104 kg/pig. Drug withdrawal prior to slaughter was 1, 3 and 5 d for pigs fed cimaterol at .25 mg/kg and 1 d for those fed cimaterol at .5 mg/kg of diet. Dietary cimaterol level influenced (quadratic, P less than .01) average daily gain during the first 42 d on test; however, daily feed intake and feed conversion ratio were not affected (P greater than .1). Pigs fed .25 mg/kg cimaterol with a 1-d drug withdrawal had 6.8, 7.7 and 13.5% less 10th rib fat depth and 11.1, 6.1 and 13.3% less P2 fat depth than those subjected to either a 3- or 5-d drug withdrawal or those fed the 0 mg/kg cimaterol diet (control), respectively. Overall, pigs fed cimaterol had 7.9% larger longissimus muscle area and 2.6% more kilograms of muscle than pigs fed the control diet. Cimaterol fed at .5 mg/kg resulted in higher (P less than .05) Warner-Bratzler shear force values and altered the proportion of saturation in some long-chain fatty acids, although the total saturated:unsaturated fat ratio was not affected. Pigs fed no cimaterol had less thaw loss (P less than .05) than did those fed other treatments.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2564388 TI - Mania secondary to amantadine treatment of neuroleptic-induced hyperprolactinemia. AB - The authors report a case of mania secondary to amantadine in a patient for whom amantadine was used to treat galactorrhea and amenorrhea. Attempts to treat the mania and endocrine symptoms simultaneously and the biological implications are discussed. PMID- 2564389 TI - Amino acid sequence deduced from a rat kidney cDNA suggests it encodes the Zn peptidase aminopeptidase N. AB - We have isolated and characterized rat kidney cDNA clones encoding a 140-kDa glycoprotein that exhibits characteristics of a cell surface Zn-peptidase. Structural features predicted for this putative kidney Zn-peptidase (KZP) are most consistent with properties previously determined for the Zn-peptidase aminopeptidase N. The deduced amino acid sequence of rat KZP is almost identical to the NH2-terminal sequence of aminopeptidase N purified from rabbit. The overall amino acid composition predicted for rat KZP is remarkably similar to that previously determined for rabbit and pig aminopeptidase N. The predicted Mr of rat kidney KZP approximates the Mr of the unglycosylated form of aminopeptidase N. The topology predicted for KZP is identical to that observed for aminopeptidase N: a short cytoplasmic domain at the NH2 terminus immediately precedes an uncleaved signal/anchor domain; a stalk region connects this membrane anchor to the extracellular, hydrophilic bulk of the protein containing catalytic sites required for Zn-peptidase activity. In addition, mRNA encoding KZP is present in tissues known to exhibit aminopeptidase N activity. Taken together with the observation that only a single gene homologous to KZP DNA is present in the rat and human genomes, these results suggest that we have established the primary structure of rat kidney aminopeptidase N. PMID- 2564390 TI - Expression of human asparagine synthetase in Escherichia coli. AB - Human asparagine synthetase was expressed in Escherichia coli. Synthesis of the enzyme was demonstrated by immunoblotting and by complementation of asparagine auxotrophy in E. coli. The recombinant enzyme was shown to have both the ammonia- and glutamine-dependent asparagine synthetase activity in vitro. Compared to asparagine synthetase isolated from beef pancreas, the one expressed in E. coli migrated at a slightly slower rate on a denaturing protein gel. In contrast with previous reports, the data obtained here strongly suggest that the active enzyme is a homodimer. The production of soluble and active enzyme was shown to be highly temperature-dependent. Expression at 37 degrees C yielded no soluble enzyme, whereas growth at 30 and 21 degrees C favored the production of soluble asparagine synthetase. The incubation temperature was also important for complementation of asparagine auxotrophy in E. coli, as growth in the absence of asparagine occurred at 30 degrees C and not at 37 degrees C. PMID- 2564391 TI - The role of glutathione in copper metabolism and toxicity. AB - Cellular copper metabolism and the mechanism of resistance to copper toxicity were investigated using a wild type hepatoma cell line (HAC) and a copper resistant cell line (HAC600) that accumulates copper and has a highly elevated level of metallothionein (MT). Of the enzymes involved in reactive oxygen metabolism, only glutathionine peroxidase was elevated (3-4-fold) in resistant cells, suggestive of an increase in the cellular flux of hydrogen peroxide. A majority of the cytoplasmic copper (greater than 60%) was isolated from both cell lines as a GSH complex. Kinetic studies of 67Cu uptake showed that GSH bound 67Cu before the metal was complexed by MT. Depletion of cellular GSH with buthionine sulfoximine inhibited the incorporation of 67Cu into MT by greater than 50%. These results support a model of copper metabolism in which the metal is complexed by GSH soon after entering the cell. The complexed metal is then transferred to MT where it is stored. This study also indicates that resistance to metal toxicity in copper-resistant hepatoma cells is due to increases in both cellular GSH and MT. Furthermore, it is suggested that elevated levels of GSH peroxidase allows cells to more efficiently accommodate an increased cellular hydrogen peroxide flux that may occur as a consequence of elevated levels of cytoplasmic copper. PMID- 2564392 TI - Isolation and characterization of the human chromosomal gene for polypeptide chain elongation factor-1 alpha. AB - The cDNA for human elongation factor-1 alpha was isolated from a cDNA library of human fibroblast cells. Using the cDNA as a probe, a number of chromosomal genes encoding the human elongation factor-1 alpha were isolated. Characterization of the clones by restriction enzyme mapping and nucleotide sequence analysis has revealed that only one of them is an active gene, whereas all of the other genes are processed pseudogenes. The active gene consists of 8 exons and 7 introns spanning about 3.5 kilobases, and the sequence of its exons is completely identical to that of the human elongation factor-1 alpha cDNA. The first non coding exon of 33 base pairs is separated by a 943-base pair intron from the coding exons. The primer extension of human elongation factor-1 alpha mRNA has indicated that the transcription of human elongation factor-1 alpha gene starts from a C residue, and a "TATA" box was found 24 base pairs upstream of the initiation site. Three and five Sp1 binding sites are present on the 5'-flanking region and the 1st intron, respectively. Furthermore, one Ap-1 binding site is located in the 1st intron. By using nuclear extracts from HeLa cells, the promoter of human elongation factor-1 alpha gene could stimulate in vitro transcription better than the adenovirus major late promoter. PMID- 2564393 TI - Regulatory properties of magnesium-dependent guanylate cyclase in Dictyostelium discoideum membranes. AB - We have characterized a magnesium-dependent guanylate cyclase in homogenates of Dictyostelium discoideum cells. 1) The enzyme shows an up to 4-fold higher cGMP synthesis in the presence of GTP analogues with half-maximal activation at about 1 microM guanosine 5'-O-(3-thio)triphosphate (GTP gamma S) or 100 microM guanosine 5'-(beta, gamma-imido)triphosphate; little or no stimulation was observed with GTP, guanosine mono- and diphosphates or with adenine nucleotides, with the exception of the ATP analogue adenosine 5'-(beta, gamma imido)triphosphate. 2) Both basal and GTP gamma S-stimulated guanylate cyclase activity were rapidly lost from homogenates as was the ability of GTP gamma S to stimulate the enzyme after cell lysis. 3) Inclusion of 25 microM GTP gamma S during cell lysis reduced the KM for GTP from 340 to 85 microM and increased the Vmax from 120 to 255 pmol/min.mg protein, as assayed in homogenates 90 s after cell lysis. 4) Besides acting as an activator, GTP gamma S was also a substrate for the enzyme with a KM = 120 microM and a Vmax = 115 pmol/min.mg protein. 5) GTP gamma S-stimulated, Mg2+-dependent guanylate cyclase was inhibited by submicromolar concentrations of Ca2+ ions, and by inositol 1,4,5-trisphosphate in the absence of Ca2+ chelators. 6) Guanylate cyclase activity was detected in both supernatant and pellet fractions after 1 min centrifugation at 10,000 x g; however, only sedimentable enzyme was stimulated by GTP gamma S. We suggest that the Mg2+-dependent guanylate cyclase identified represents the enzyme that in intact cells is regulated via cell surface receptors, and we propose that guanine nucleotides are allosteric activators of this enzyme and that Ca2+ ions play a role in the maintenance of the enzyme in its basal state. PMID- 2564394 TI - Characterization of an endoprotease from rat small intestinal mucosal secretory granules which generates somatostatin-28 from prosomatostatin by cleavage after a single arginine residue. AB - We have extracted, characterized, and partially purified an enzyme from secretory granules from rat small intestinal mucosa which cleaves a synthetic prosomatostatin substrate on the carboxyl side of a single arginine residue. This substrate Leu-Gln-Arg-Ser-Ala-Asn-Ser-NH2 contains the monobasic site at which mammalian prosomatostatin is cleaved in vivo to generate somatostatin-28. This activity was released from the granules by osmotic shock followed by extraction with 500 mM KCl. The enzyme had a molecular weight of about 55,000, a pH optimum of about 7.5, and a Km for the synthetic substrate of 20 microM. It was partially inhibited by diisopropyl fluorophosphate, phenylmethanesulfonyl fluoride, iodoacetate, soybean trypsin inhibitor, and EDTA. It was also very sensitive to aprotinin (complete inhibition at 25 micrograms/ml) but was not inhibited by bestatin, pepstatin, or p-chloromercuribenzoate. This endoprotease was unable to cleave three small trypsin and kallikrein substrates (N alpha-benzoyl-L-arginine ethyl ester, N alpha-benzoyl-DL-arginine p-nitroanilide, and N alpha-benzoyl-L arginine 7-amido-4-methylcoumarin). It was unable to cleave either the Arg-Asp bond in CCK 12 or the Arg-Glu and Arg-Met bonds of synthetic peptides corresponding to sequences of anglerfish prosomatostatin II situated upstream from the somatostatin-28 domain. These observations together suggest that adjacent amino acids play a role in determining the conformational specificity of the monobasic cleavage. This soluble enzyme was also able to cleave three synthetic substrates containing dibasic residues (Arg-Lys or Lys-Arg) on the carboxyl side of the arginine, although it did so less rapidly than at the monobasic cleavage sites. When incubated with partially purified prosomatostatin from anglerfish pancreas, significant quantities of somatostatin-28 II were produced. All these cleavages were completely blocked by preincubation with aprotinin. Although further work is required to clarify the physiological role of this enzyme, it appears, in view of its catalytic properties, this endoprotease could be involved in the conversion of prosomatostatin to somatostatin-28 in intestine mucosal secretory cells. PMID- 2564395 TI - Vasoactive intestinal peptide stimulates long-chain fatty acid oxidation and inhibits acetyl-coenzyme A carboxylase activity in isolated rat enterocytes. AB - The effects of vasoactive intestinal peptide (VIP) on fatty acid oxidation in isolated rat enterocytes were investigated. VIP (10(-7) M) increased more than 2 fold the production of 14CO2 from [U-14C]palmitate. This effect was dose dependent (K0.5 = 5.10(-11) M) and appeared to be related to the stimulation of cAMP production since it was mimicked by forskolin (10(-4) M). VIP also stimulated oxygen consumption of the cells, an effect accounted for by the stimulation of the oxidation of both exogenous added palmitate (0.12 mM) and endogenous fatty acids produced by lipolysis. VIP appeared to specifically enhance the oxidation of long-chain fatty acids since its effects were counteracted by 5.10(-5) M sodium 2-[6-(chlorophenoxy)hexyl]oxirane-2 carboxylate, a potent inhibitor of carnitine palmitoyltransferase 1, and since VIP did not affect cell respiration in the presence of octanoate. These results suggested that VIP stimulated long-chain fatty acid oxidation by increasing their translocation into the mitochondria. Therefore, we examined the effect of VIP on the activity of acetyl-coenzyme A carboxylase, the enzyme responsible for the biosynthesis of malonyl-CoA, a physiological inhibitor of carnitine acyltransferase 1. VIP produced an acute, dose-dependent (Ki = 3.10(-11) M), 90% inhibition of acetyl-coenzyme A carboxylase activity. These results allow us to elucidate the mechanism of the recently reported inhibitory effect of VIP on glucose oxidation (Vidal, H., Comte, B., Beylot, M., and Riou, J. P. (1988) J. Biol. Chem. 263, 9206-9211) and demonstrate for the first time that balance between fatty acids and glucose as energetic fuels is under neurohormonal control in isolated rat enterocytes. PMID- 2564396 TI - Alpha 2-adrenergic receptor stimulation mobilizes intracellular Ca2+ in human erythroleukemia cells. AB - Human erythroleukemia cells are a model system for studies of alpha 2-adrenergic receptors and their coupling to inhibition of adenylate cyclase (McKernan, R. M., Howard, M. J., Motulsky, H. J., and Insel, P. A. (1987) Mol. Pharmacol. 32, 258 265). Using Fura-2, we show that alpha 2-adrenergic receptor stimulation also increases intracellular Ca2+ in these cells by 80-250 nM. Although epinephrine only inhibited forskolin-stimulated cAMP generation when beta-adrenergic receptors were blocked, the Ca2+ increase was not affected by beta-adrenergic receptor blockade. The Ca2+ increase was not affected by forskolin or 8-bromo cAMP. Thus, alpha 2-adrenergic receptors independently couple to elevation of intracellular Ca2+ and adenylate cyclase inhibition. Chelating all extracellular Ca2+ did not reduce the response, demonstrating mobilization of intracellular, rather than influx of extracellular Ca2+. The epinephrine-stimulated Ca2+ mobilization occurred prior to any detectable increase in inositol-(1,4,5) trisphosphate. It was abolished by pretreatment with pertussis toxin (which blocks some G protein-mediated processes), but not by aspirin and indomethacin (which inhibit cyclooxygenase), nordihydroguaiaretic acid (which inhibits lipoxygenase), or Na+-free buffer (to block any Na+H+ exchange). We conclude, therefore, that alpha 2-adrenergic receptors on human erythroleukemia cells couple to mobilization of intracellular Ca2+ via a (pertussis toxin-sensitive) G protein-mediated mechanism that is independent of inhibition of adenylate cyclase. PMID- 2564397 TI - Transformation-sensitive and growth-related changes of protein synthesis in REF52 cells. A two-dimensional gel analysis of SV40-, adenovirus-, and Kirsten murine sarcoma virus-transformed rat cells using the REF52 protein database. AB - Two-dimensional gels of normal and virally transformed REF52 cells have been quantified and compared using the QUEST system for construction and analysis of protein databases. The REF52 protein map is based on more than 1600 high quality spots, and the relative amounts of these proteins are studied in 79 gels representing 12 major experiments. REF52 cells transformed by SV40, adenovirus, and Kirsten murine sarcoma virus (KiMSV) are compared to normal REF52 cells at several stages of growth from low density to confluence and after refeeding confluent cells. In addition, early (1-4 h) and late (21-24 h) responses to serum stimulation were measured in normal, SV40-and adenovirus-transformed cells. The database has been analyzed with respect to 1) known marker proteins and protein sets, 2) global comparison of protein patterns, and 3) selection of unknown spots which have interesting patterns of regulation. For the marker proteins, which include the tropomyosin family and the proliferation-sensitive nuclear antigen, new aspects of regulation by growth and transformation have been revealed. Proliferation-sensitive nuclear antigen, a protein known to be involved in DNA synthesis, is growth-regulated in normal cells and overexpressed in some SV40- and adenovirus-transformed cells. Global comparisons reveal no overall correlation between growth-regulated changes and transformation-induced changes; however, a set of 26 coregulated proteins, including proliferation-sensitive nuclear antigen, was found to be overexpressed in REF52 cells transformed by SV40 or adenovirus. These proteins are synthesized at rates that correlate with the rate of cell proliferation in REF52 and Kirsten murine sarcoma virus-transformed cells but, in SV40- and adenovirus-transformed cells, these proteins are synthesized at high levels independent of the rate of growth. These data suggest that the transforming proteins of SV40 and adenovirus share a function that results in deregulation of the genes coding for a class of cell cycle-regulated proteins. PMID- 2564399 TI - Beta-adrenergic control of cell volume and chloride transport in an established rat submandibular cell line. AB - Rat submandibular cells treated with methylcholanthrene are able to be propagated in continuous culture while retaining beta-adrenergic responsiveness. A specific clone, RSMT-A5, has been isolated and studied in detail. RSMT-A5 cells possess beta-adrenergic receptors (BARS) as judged by [3H]-dihydroalprenolol ([3H]-DHA) binding studies. [3H]-DHA binds to RSMT-A5 membranes in a specific and saturable manner with respect to time and [3H]-DHA concentration. Specific binding is saturable within three min of incubation, and a Scatchard analysis reveals a single class of high affinity binding sites with an equilibrium dissociation constant of 0.62 +/- 0.03 nM and a receptor density of 101 +/- 4 fmole/mg protein. Antagonist competition studies indicate that the BARs are primarily of the beta 2-subtype. The BARs are functional since isoproterenol stimulation results in an increased intracellular cAMP content, marked morphological change, and decreased cell volume and chloride content. These same responses can be evoked by treating RSMT-A5 cells with 8-bromo-cAMP. Ion transport inhibitors such as bumetanide (an inhibitor of Na/K/Cl cotransport), SITS and DIDS (inhibitors of chloride-bicarbonate exchange), amiloride (an inhibitor of Na-H exchange), ouabain (an inhibitor of Na/K-ATPase), and dipyridamole and 9-anthracene carboxylic acid (chloride channel blockers) fail to inhibit the isoproterenol stimulated change in chloride content. The effects of either isoproterenol or 8 bromo-cAMP on both chloride content and cell volume can be inhibited by the chloride channel blocker N-phenylanthranilic acid, however. Taken together, our results indicate that RSMT-A5 cells possess a beta-adrenergic receptor system which controls intracellular volume and chloride content by modulating transport processes that are 1) cAMP-responsive and 2) inhibitable by the putative chloride channel blocker N-phenylanthranilic acid. PMID- 2564398 TI - Endocytosis from coated pits of Shiga toxin: a glycolipid-binding protein from Shigella dysenteriae 1. AB - Evidence is presented that endocytosis is involved in the transport to the cytosol of the cytotoxin from Shigella dysenteriae 1, Shiga toxin, which acts by removal of a single adenine residue in 28-S ribosomal RNA. Inhibition of endocytosis by ATP depletion of the cells prevented toxin uptake. Exposure of HeLa S3 and Vero cells to toxin at low extracellular pH, where translocation to the cytosol, but not endocytosis is inhibited, allowed the toxin to accumulate in a compartment where it was protected against antibodies to the toxin. Upon transfer of the cells to normal medium endocytosed toxin entered the cytosol. Electron microscopical studies of cells exposed at 0 degrees C to a toxin horseradish peroxidase (HRP) conjugate, or to unconjugated toxin followed by horse antitoxin antibodies and then protein G-gold, revealed that the Shiga toxin binding sites were randomly distributed on the cell surface, without any preference to, for example, coated pits. In contrast, when cells were exposed to toxin at 37 degrees C, the binding sites were preferentially localized in coated pits. The Shiga-HRP conjugate was also seen in endosomes, lysosomes, and in the Golgi region. Endocytosis by the coated pit/coated vesicle pathway was selectively inhibited by acidification of the cytosol. Under these conditions, both the uptake of toxin-HRP conjugates and intoxication of the cells were inhibited. Evidence from the literature as well as our own results suggest that Shiga toxin binding sites are glycolipids. Thus, Shiga toxin appears to be the first example of a lipid-binding ligand that is endocytosed from coated pits. PMID- 2564400 TI - Effects of temperature on the degradation and biliary secretion of asialoorosomucoid by the perfused rat liver: evidence for two intracellular pathways. AB - We have utilized the in situ perfused rat liver under nonrecirculating conditions to examine the effect of temperature on the metabolism and biliary secretion of [125I]-asialoorosomucid (ASOR). In this manner we were able to follow the fate of a single round of internalized ligand. In control livers perfused at 37 degrees C, approximately 50% of [125I]-ASOR injected into the portal vein was extracted on first pass. Five minutes after the injection, radioactivity, which had been extracted initially, began to appear in the hepatic venous effluent. Within 25 min, 50% of the initially extracted radioactivity was released into the perfusion medium; the bulk of this radioactivity (greater than 95%) was soluble in trichloroacetic acid. In livers perfused at temperatures slightly less than 37 degrees C (30-35 degrees C), first-pass extraction of [125I]-ASOR was similar to that observed at 37 degrees C. However, a severalfold decrease in the rate of release of radioactivity from the liver into the perfusion medium was noted at the lower perfusion temperatures; whereas greater than 50% of the initially extracted radioactivity was released within 30 min from livers perfused at 37 degrees C, only 5% was released at 30 degrees C. At the lower perfusion temperature, a larger proportion of the released radioactivity was acid precipitable (24% vs. 5%). Some radioactivity also was recovered in the bile; of the total amount of radioactivity released from the liver in 30 min at 37 degrees C, approximately 5% was directed into the bile. At lower temperatures of perfusion, a greater fraction of the radioactivity that was released from the liver was directed into the bile (20% at 30 degrees C vs. 5% at 37 degrees C). The data imply that the endosomal pathway to the lysosome is highly sensitive to slight reductions in temperature while the transcytotic route into bile is less sensitive. Lower temperatures might prolong the residence time of ASOR in the prelysosomal endosomal compartments, and thereby increase the likelihood that undegraded ligand will be returned to the blood or be missorted into bile. PMID- 2564401 TI - Concentration dependence of class III and beta-adrenergic blocking effects of sotalol in anesthetized dogs. AB - Sotalol is unique among beta-adrenergic blocking drugs in possessing significant class III antiarrhythmic actions. The present study was designed to assess the relative concentration dependence of beta-blocking and class III actions of sotalol and to relate the findings to concentrations achieved during oral sotalol therapy in humans. Measurements were made in anesthetized dogs under control conditions, and then in the presence of a series of stable sotalol plasma concentrations produced by sequential loading and maintenance infusions. Beta blocking effects of sotalol, determined by attenuation of the chronotropic actions of isoproterenol, were seen at the lowest dose used. Increases in atrial and ventricular refractory periods (observed in dogs with autonomic blockade to exclude beta-receptor-mediated or reflex autonomic effects) required much larger doses of sotalol. Half-maximal beta-blocking effects occurred at an average sotalol concentration of 0.8 +/- 0.3 mg/liter, an order of magnitude lower than the concentrations required for half-maximal effects on atrial (6.9 +/- 1.2 mg/liter, p less than 0.01) and ventricular (6.8 +/- 2.8 mg/liter, p less than 0.05) refractoriness. These results show that substantially higher concentrations are needed for the class III effects of sotalol than for its beta-blocking action. These pharmacodynamic differences need to be considered in evaluating the antiarrhythmic efficacy and mechanisms of this unusual drug. PMID- 2564402 TI - Comparative effects of carbohydrate restriction vs starvation on biochemical parameters related to neurotransmitters in rat. AB - Adult rats were submitted to a 4-day starvation period or maintained on a 50% carbohydrate-restricted diet for 8 consecutive days to obtain a body weight loss of 20-30%. Serum dopamine-beta-hydroxylase (DBH) activity and amino acids content were measured as well as brain tryptophan and tyrosine levels. Moreover, brain serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), noradrenaline (NA), and dopamine (DA) contents were assayed in five brain areas. In 4-day starved and 8 day carbohydrate-restricted rats, the serum tyrosine and total tryptophan contents as well as tyrosine to the sum of six neutral amino acids ratios were lowered. Moreover, in these groups, free tryptophan to the sum of six neutral amino acids ratio remained normal and serum DBH activity increased. In the brain, to a decreased tyrosine content observed in 4-day starved and 8-day carbohydrate restricted rats corresponded a high DA to NA ratio in the hypothalamus, thalamus, and raphe nuclei, thus suggesting a low DA utilization whereas a low DA to NA ratio was found in the neostriatum. On the other hand, brain tryptophan content was decreased in 4-day starved rats and increased in 8-day carbohydrate restricted rats. In the former group, a high 5-HT to 5-HIAA ratio characteristic of a low 5-HT utilization was found in the hypothalamus and neostriatum whereas in the latter group a significant decrease in this ratio was only observed in the thalamus. These results suggest that the biochemical response to starvation vs carbohydrate restriction can be differentiated on neurochemical and neuroanatomical bases. PMID- 2564403 TI - Dr. Sullivan's response to Dr. Toogood's editorial. PMID- 2564404 TI - Ultrastructural localization of neuropeptides and GABA in rat dorsal horn: a comparison of different immunogold labeling techniques. AB - Several immunogold techniques were used to determine the ultrastructural localization of calcitonin gene-related peptide (CGRP), tachykinin, somatostatin, and gamma-amino-butyric acid (GABA) immunoreactivity in the dorsal horn of rat spinal cord. The immunocytochemical reactions were carried out directly on ultrathin sections from non-osmicated frozen tissue, non-osmicated low temperature-embedded (Lowicryl K4M) tissue, and osmicated epoxy-embedded material. Preservation of ultrastructural morphology and immuno-labeling efficiency were compared. Morphology of subcellular organelles was relatively good in ultra-thin frozen sections, which showed the highest immunoreactivity. However, only very small samples of tissue could be examined. Although there was relatively good immunolabeling in the Lowicryl K4M-embedded tissue, the ultrastructure of the neuropil, and particularly that of synapses, was poorly maintained. In contrast, the osmicated epoxy-embedded material offered optimal morphological preservation together with accurate subcellular localization of all antigens under study. The latter approach thus enabled clear visualization of CGRP, tachykinin, and somatostatin immunoreactivity restricted to large dense cored vesicles (90-150 nm diameter) in many axonal and synaptic profiles in the superficial layers of the dorsal horn. CGRP- and tachykinin-positive profiles were also present in the tract of Lissauer. GABA immunoreactivity was present mainly in axons and terminals, and less frequently in somatic and dendritic profiles. In terminals, which often formed symmetrical synapses on immunonegative dendritic profiles, it was associated with small (30-60 nm diameter) clear vesicles and mitochondria. Double immunolabeling was possible on all preparations, but the osmicated, epoxy-embedded material clearly showed co localization of peptides, especially of CGRP and tachykinins, within the same dense-cored vesicles in axonal fibers and/or terminals. On the other hand, peptide and GABA immunoreactivity were consistently seen in different nerve profiles. In a few cases, GABAnergic terminals were seen to synapse on tachykinin positive fibers. PMID- 2564405 TI - Restriction fragment length polymorphisms for the renin gene in Dahl rats. AB - Genomic DNAs from inbred Dahl salt-hypertension sensitive (S) and inbred Dahl salt-hypertension resistant (R) rats were examined for restriction fragment length polymorphisms (RFLPs) using a rat renin cDNA probe. Eight of the 28 restriction enzymes used yielded polymorphic fragments between S and R strains. This suggests a major structural difference in or near the renin gene of Dahl rats. Some, but not all, of the polymorphisms are compatible with an insertion/deletion mutation of about 1.1 kb in size. The ratio of renin gene copy numbers for S to R was found to be 1, showing that the RFLPs are not likely to be due to gene duplication. PMID- 2564406 TI - The role of Ca2+ and Mg2+ in the cytotoxic T lymphocyte reaction and in the secretion of N alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester-serine esterase by human T cell clones. AB - Human T cell clones contain enzymes that can cleave the substrate N-alpha benzyloxycarbonyl-L-lysine thiobenzyl ester (BLT). All CTL clones tested in this study secreted BLT-serine esterase activity, whereas only one of three tested non cytolytic T cell clones secreted this enzymatic activity upon Ag-specific activation. BLT-serine esterase secretion could also be induced by the Fc gamma+ target cell Daudi in the presence of mAb specific for the TCR/CD3 complex, CD2, or the T cell activation Ag Tp 103. In addition, anti-CD3 and a mitogenic combination of anti-CD2 mAb, induced secretion of BLT-serine esterase in the absence of target cells, whereas anti-Tp 103 failed to do so. The secreted BLT serine esterase activity induced by the various ligands was inhibited by the serine esterase inhibitors PMSF and m-ABA, but not by N-alpha-p-tosyl-L-lysine chloromethyl ketone. Significant BLT-serine esterase activity was induced by target cells or soluble anti-CD3 in the absence of extracellular Ca2+ ions, provided that extracellular Mg2+ ions were present. The cytotoxic activities by the human CTL clones were completely blocked under these conditions. All ligands that induced BLT-serine esterase secretion in the absence of extracellular Ca2+, induced a transient rise of intracellular Ca2+. Soluble anti-CD3 mAb did not induce a transient rise in intracellular Ca2+ or secretion of BLT serine esterase in CTL preincubated for 2 h with 5 mM EGTA. These findings indicate that mobilization of intracellular Ca2+ in human CTL clones is required for induction of secretion of BLT-serine esterase. PMID- 2564407 TI - Endothelial-leukocyte adhesion molecule-1-dependent and leukocyte (CD11/CD18) dependent mechanisms contribute to polymorphonuclear leukocyte adhesion to cytokine-activated human vascular endothelium. AB - We have examined the contributions of endothelial-leukocyte adhesion molecule-1 (ELAM-1) and the complex of leukocyte surface adhesion molecules designated CD11/CD18 to the adhesion of human polymorphonuclear leukocytes (PMN) to cultured human endothelial cells (HEC), activated by rIL-1 beta for 4 or 24 h. Inhibition of PMN attachment to IL-1-activated HEC was measured in a quantitative in vitro monolayer adhesion assay, after treatment with mAb directed to ELAM-1 (mAb H18/17), and to CD11a (mAb L11), CD11b (mAb 44), CD11c (mAb L29), and CD18 (mAb 10F12), alone or in combination. Pretreatment of activated HEC with mAb H18/7 inhibited PMN adhesion by 47 +/- 8% whereas control mAb had no effect. CD11/CD18 directed mAb significantly blocked PMN adhesion to activated HEC (anti-CD11a, 40 +/- 3%; anti-CD11b, 34 +/- 4%; anti-CD18, 78+/- 6% inhibition). The combination of mAb H18/7 and each of the various anti-CD11/CD18 mAb resulted in greater inhibition of PMN adhesion than any Mab alone. After 24 h of rIL-1 beta treatment, when ELAM-1 was markedly decreased but elevated PMN adhesion was still observed, mAb H18/7 had no effect on PMN adhesion. At this time, CD11/CD18 dependent adhesive mechanisms predominated and a CD11c-dependent mechanism became apparent (anti-CD11a, 67 +/- 4% inhibition; anti-CD11b, 45 +/- 9%; anti-CD11c, 26 +/- 6%; anti-CD18, 97 +/- 1%). In summary, PMN adhesion to IL-1-activated HEC involves both CD11/CD18-dependent mechanisms and an ELAM-1-dependent mechanism, and the relative contribution of these varies at different times of IL-1-induced HEC activation. The additive blocking observed at 4 h with mAb H18/7 in combination with CD11/CD18-directed Mab implies that members of the CD11/CD18 complex do not function as an obligate ligand(s) for ELAM-1. PMID- 2564408 TI - The role of natural suppressor and natural killer activities in resistance to hemopoietic transplantation in unirradiated hosts. AB - We report here that bone marrow stem cell engraftment in unirradiated hosts correlates with levels of natural suppressor (NS) activity in the host at the time of transplantation. This is shown in the antibody-facilitated murine chimeras, in which conditioning consists of a single injection of anti-host MHC antibody, which results in long term hemopoietic engraftment in P----F1 and syngeneic donor-host combinations. The data establish that, in two independent situations, engraftment is reduced in hosts with elevated NS activity. Resistance to engraftment in antibody-conditioned adult hosts is increased by prior administration of CFA, which also increases NS activity. Likewise, neonatal animals, which are highly resistant to antibody-facilitated engraftment, exhibit a spontaneously-increased level of NS activity. This resistance declines with the ontogenic waning of splenic NS activity. Conversely, administration of facilitating antibody decreases host bone marrow NS activity, while anti-MHC antibodies that fail to facilitate engraftment do not reduce it. NK activity, on the other hand, correlates poorly with resistance or susceptibility to marrow engraftment in these situations. These results suggest that immunoregulatory functions associated with hemopoiesis may control engraftment of donor stem cells in unirradiated hosts. PMID- 2564409 TI - Fluorescence intensity analysis through simplex optimization in flow cytometry. AB - Fluorescence intensity analysis in flow cytometric surface immunophenotyping has recently been appreciated in clinical applications. A curve fitting method to estimate the mean and SD values of fluorescence intensity is described in this report. A Gaussian distribution is aimed to be adapted for a specified distribution in logarithmically scaled histogram data through the simplex optimization, one of the non-linear least squares methods. In comparison with the conventional methods which include the detection of peak point and the direct calculation, this fitting method has demonstrated exceeding precisions in the estimation of both parameters with limited involved cell counts in typical lymphocytic phenotyping. The actual estimation for a precise SD value will develop the quality control approaches based on the fluorescence intensity analysis. While this method is not suitable for distributions that involve extremely small cell counts or that deviate markedly from a symmetric Gaussian, it has additional advantages of loose requirements, namely, narrow fitting regions, ordinarily small cell counts, practical computational periods and a simple programming. PMID- 2564410 TI - [Clinical usefulness of serum sialyl Le(x)-i measurement in patients with ovarian cancer]. AB - Sialyl Le(X)-i (Sialyl SSEA-1, SLX) is one of the type 2 chain carbohydrate antigens, which is defined by a monoclonal antibody FH-6. The clinical usefulness of the measurement of serum Sialyl LeX-i levels in the follow-up study of outpatient with ovarian cancer was examined for the early detection of recurrence as the serodiagnostic test. Elevated serum Sialy Le(X)-i levels (more than 38 unit/ml) were observed before treatment in 7 of 12 patients with a good prognosis (group A) and in 15 of 26 patients with a poor prognosis (group B). Six (85.7%) in 7 patients with elevated serum Sialyl Le(X)-i levels in group A decreased to the normal range after treatment, whereas 3 (20.0%) in 15 patients with positive serum Sialyl Le(X)-i levels in group B decreased below 38 unit/ml a after treatment. In 9 (34.6%) in group B, elevated serum Sialyl Le(X)-i levels were observed with an average 3.1 weeks before clinical evidence of recurrence. With tumor progression, serum Sialyl Le(X)-i levels also rose in 25 (96.2%) in group B. In 4 (15.4%) in group B, serum Sialyl Le(X)-i levels were a useful tumor marker compared with others including CA 125, TPA, and CEA. Consequently, the measurement of serum Sialyl Le(X)-i levels may be useful to monitor the condition of the disease, presume progression, and detect recurrence early in outpatients with ovarian cancer. PMID- 2564411 TI - Neutrophil membrane sulfhydryl groups are involved in stimulated neutrophil adherence to endothelium. AB - We investigated the role of membrane sulfhydryl groups in adherence of stimulated polymorphonuclear neutrophils to cultured endothelial cells. Treatment of neutrophils with p-chloromercuriphenyl sulfonate (PCMPS), a slowly penetrating sulfhydryl reagent, inhibited phorbol myristate acetate (PMA)- or calcium ionophore A23187-stimulated adherence to cultured human or bovine endothelial cells. At concentrations which completely blocked PMA-stimulated adherence, PCMPS did not cause release of lactic dehydrogenase, inhibit PMA-mediated degranulation or hydrogen peroxide production, or prevent the PMA-induced increased surface expression of CD11b/CD 18 (Mac-1). Coincubation with a competing reduced sulfhydryl compound protected neutrophils from inhibition of PMA-stimulated adherence by PCMPS, whereas coincubation with an oxidized sulfhydryl compound did not. Monobromotrimethylammoniobimane, a nonpenetrating sulfhydryl reagent that is structurally unrelated to PCMPS, also inhibited stimulated neutrophil adherence to endothelium cultures. We conclude that stimulated neutrophil adherence to endothelium involves neutrophil membrane protein sulfhydryl groups. PMID- 2564412 TI - Selenium and the immune response: 2. Enhancement of murine cytotoxic T-lymphocyte and natural killer cell cytotoxicity in vivo. AB - An inverse correlation between cancer incidence and dietary intake of the trace mineral element selenium has been well established in epidemiological and experimental studies. The mechanisms for this chemoprotective effect are unresolved. Much attention has been focused on the antiproliferative effects of selenium on various normal and neoplastic cell types. However, dietary selenium supplementation can also enhance the expression of various humoral and cellular immune responses. In examining the effects of dietary selenium on cell-mediated immunity in mice, we observed that selenium supplementation caused the enhanced expression of spontaneous natural killer (NK) cytotoxicity in spleen cells and of specific cytotoxic T-lymphocyte (CTL) cytotoxicity in peritoneal exudate cells (PEC). NK activity of spleen-cell suspensions from selenium-supplemented mice increased an average of 70% over that of the control group (basal diet). Cytotoxic activity of PEC from mice injected with tumors intraperitoneally peaked earlier in selenium-supplemented animals, and the appearance of cells staining positively for Thy 1.2 surface antigen in selenium-supplemented animals also preceded the values observed in control animals. We propose here that enhancement of in vivo cytotoxic mechanisms, is likely to act synergistically with tumor growth inhibition in the reduction of tumor incidence associated with selenium intake. PMID- 2564413 TI - The tumor-rejection antigens of the 1591 ultraviolet fibrosarcoma. Potential origin and evolutionary implications. AB - Previously, we cloned and sequenced the three novel MHC class I genes expressed by the C3H UV fibrosarcoma, 1591. We have extended the analysis of the polymorphic nature of these genes relative to the C3H strain. Scattered nucleotide differences among the tumor genes as compared with the C3H H-2 and Qa sequences make it highly unlikely that the novel tumor genes were generated by recombination between endogenous C3H sequences. Given that two of the tumor clones, A149 and A166, are remarkably similar in amino acid and DNA sequence to H 2Lq and H-2Dq, respectively, we also examined the 1591 RP2 and GUS loci for evidence of polymorphism. Compared with C3H and B10.AKM, 1591 appears to be heterozygous at each of these loci, consistent with an H-2q origin for the two novel 1591 class I genes. Interestingly, the third tumor gene, designated A216, shares certain characteristics with the H-2Ks antigen, reminiscent of the naturally occurring combination of H-2Ks, H-2Dq, and H-2Lq antigens found in some Swiss mouse strains. As a result, we propose that the non-C3H/HeN characteristics displayed by the 1591 tumor point to a non-C3H origin for the novel tumor class I genes of 1591. PMID- 2564414 TI - Structure of the human CR1 gene. Molecular basis of the structural and quantitative polymorphisms and identification of a new CR1-like allele. AB - Structural and quantitative polymorphisms have been described in human CR1. In the former, the S allotype is larger than the F allotype by 40-50 kD, the size of a long homologous repeat (LHR). In the latter, homozygotes for a 7.4-kb Hind III fragment express fourfold more CR1 per erythrocyte than do homozygotes for the allelic 6.9-kb restriction fragment. The basis for these genomic polymorphisms has been determined by restriction mapping the entire S allele and part of the F allele. The S allele is 158 kb and contains 5 LHRs of 20-30 kb, designated -A, B/A, -B, -C, and -D, respectively, 5' to 3'. Extensive homology was found among the LHRs in their restriction maps, exon organization, and the coding and noncoding sequences. The presence of LHR-B/A in the S allele but not in the F allele accounts for the longer transcripts and polypeptide associated with the former allotype. At least 42 exons are present in the S allele, with distinct exons for the leader sequence, the transmembrane and cytoplasmic regions and most of the SCRs comprising the extracellular portion of CR1. Consistent with the mapping of the ligand binding site to the first two SCRs in each LHR, the second SCRs in LHR-A, -B/A, -B, and -C are encoded by two exons, reflecting a specialized function for this unit. The allelic 7.4/6.9-kb Hind III fragments extend from the 3' region of LHR-C to LHR-D. The 6.9-kb restriction fragment is the result of a new Hind III site generated by a single base change in the intron between the exons encoding the second SCR of LHR-D. A second cluster of genomic clones has been identified by hybridization to CR1 probes. Although they contain regions of hybridization to the cDNA and genomic probes derived from CR1, these cannot be overlapped with the structural gene owing to their distinct restriction maps. Three genomic polymorphisms previously identified by CR1 cDNA probes map to this region. These additional clones may represent part of a duplicated allele located nearby within the CR1 locus. PMID- 2564415 TI - Site-restricted persistent cytomegalovirus infection after selective long-term depletion of CD4+ T lymphocytes. AB - We have established a murine model system for exploring the ability of a CD4 subset-deficient host to cope with cytomegalovirus infection, and reported three findings. First, an antiviral response of the CD8 subset of T lymphocytes could be not only initiated but also maintained for a long period of time despite a continued absence of the CD4 subset, whereas the production of antiviral antibody proved strictly dependent upon help provided by the CD4 subset. Second, no function in the defense against infection could be ascribed as yet to CD4-CD8- T lymphocytes, which were seen to accumulate to a new subset as a result of depletion of the CD4 subset. This newly arising subset did not substitute for CD4+ T lymphocytes in providing help to B lymphocytes, and was also not effective in controlling the spread of virus in host tissues. As long as a function of these cells in the generation and maintenance of a CD8 subset-mediated response is not disproved, caution is indicated with concern to an autonomy of the CD8 subset. Third, even though with delay, the CD8+ effector cells raised in the CD4 subset-deficient host were able of clear vital tissues from productive infection and to restrict asymptomatic, persistent infection to acinar glandular epithelial cells in salivary gland tissue. PMID- 2564416 TI - Human lymphocytes bearing T cell receptor gamma/delta are phenotypically diverse and evenly distributed throughout the lymphoid system. AB - A direct quantitative and phenotypic cytofluorographic analysis of TCR gamma/delta+ lymphocytes as well as an immunohistologic study of their tissue distribution and microanatomy was made possible by the availability of two mAbs (anti-TCR-delta 1 and anti-C gamma M1) specific for framework determinants on human TCR gamma and delta chains, respectively. TCR-gamma/delta+ lymphocytes, ranging between greater than 0.5 and 16% of CD3+ cells, were found in fetal and postnatal thymus, fetal and adult peripheral lymphoid organs, and adult peripheral blood. While TCR-gamma/delta+ lymphocytes comprised a small subpopulation of T cells (mean, approximately 4%) occasionally greater than 10 16% of CD3+ cells expressed TCR-gamma/delta. Virtually all TCR-gamma/delta+ thymocytes/lymphocytes expressed CD7, CD2, and CD5 but were heterogeneous with respect to their expression of CD1, CD4, CD8, CD28, CD11b, CD16, and Leu-7. Human TCR-gamma/delta+ cells populate both organized lymphoid tissues (thymus, tonsil, lymphnode, and spleen) as well as the gut- and skin-associated lymphoid systems at similar frequencies without obvious tropism for epithelial microenvironments. TCR-gamma/delta+ lymphocytes tend to be located within a given organ wherever TCR alpha/beta+ lymphocytes are found. This study shows that TCR-gamma/delta+ lymphocytes constitute a small but numerically important, phenotypically diverse T cell population distributed throughout the body. These results support the concept that TCR-gamma/delta+ cells comprise a distinct, functionally heterogeneous, mature T cell sublineage that may substantially broaden the T cell repertoire at all immunologically relevant sites. PMID- 2564418 TI - A subset of memory CD4+ helper T lymphocytes identified by expression of Pgp-1. AB - The Pgp-1 glycoprotein (Ly-24 antigen) is acquired by mature murine T lymphocytes at the time of primary antigen stimulation Pgp-1 was previously shown to be a useful cell surface marker for distinguishing antigen-specific memory CD8+ T lymphocytes after immunization. Here we demonstrate that this observation extends to CD4+ T lymphocytes. Antigen-specific CD4+ T cells in mice immunized with sperm whale myoglobin or keyhole limpet hemocyanin were contained nearly exclusively in the minor Pgp-1+ subset. PMID- 2564417 TI - Human cytomegalovirus induces stage-specific embryonic antigen 1 in differentiating human teratocarcinoma cells and fibroblasts. AB - Cell surface expression of stage specific embryonic antigen 1 (SSEA-1), or Lex (III3 FucnLC4), was induced in differentiated human teratocarcinoma cells and in human diploid fibroblasts 3-6 d after infection with human cytomegalovirus (HCMV). In parallel, fucosylated lactoseries glycolipids bearing the SSEA-1/Lex epitope were readily detected in the infected cells but not in the uninfected cells. HCMV infection also results in altered expression of several glycosyltransferases. SSEA-1/Lex induction is probably a consequence of both increased expression of beta 1----3N-acetylglucosaminyltransferase, which catalyzes the rate-limiting step in lactoseries core chain synthesis, and subtle alterations in the relative competition for common precursor structures at key points in the biosynthetic pathway. Since SSEA-1 has been suggested to play a role in some morphogenetic cell-cell interactions during embryonic development, the induction of this antigen at inappropriate times might provide one mechanism whereby intrauterine infection with HCMV can damage the developing fetal nervous system. PMID- 2564419 TI - Genetic organization of complement receptor-related genes in the mouse. AB - Using an interspecific cross, gene linkage relationships among members of the murine complement receptor-related genes, C4bp, Cfh, Mcry, and Mcr2, were analyzed by segregation of RFLP in 200 mice. The human homologues of these genes are tightly linked, composing the RCA locus, which maps to human chromosome (Chr.)1q32, within a large linkage group conserved between human Chr.1q21-32 and mouse Chr.1. RFLP associated with C4bp and Cfh map within this conserved linkage group; Cfh is located 9 cM telomeric to C4bp, which is consistent with linkage data for their human homologues. Mcry and Mcr2, while tightly linked, are located outside the conserved group, 40 cM telomeric to C4bp. These data suggest that a translocation or inversion occurred within the RCA family during the evolution of the mouse, defining a breakpoint of this large conserved linkage group. PMID- 2564420 TI - Interaction between S-100 proteins and steady-state and taxol-stabilized microtubules in vitro. AB - S-100 proteins are a group of three 21-kilodalton, acidic, Ca2+-binding proteins of the "E-F hand" type shown to regulate several cell activities, including microtubule (MT) assembly-disassembly. We show here that S-100 proteins interact with MTs assembled from either whole microtubule protein or purified tubulin, both in the absence and in the presence of the MT-stabilizing drug taxol. Evidence for the binding of S-100 to MTs comes from both kinetic (turbidimetric) and binding studies. Kinetically, S-100 enhances the disassembly of steady-state MTs in the presence of high concentrations of colchicine or vinblastine at 10 microM free Ca2+ and disassembles taxol-stabilized MTs at high Ca2+ concentrations. Experiments performed using 125I-labeled S-100 show that S-100 binds Ca2+ independently to a single set of sites on taxol-stabilized MTs assembled from pure tubulin with an affinity of 6 x 10(-5) M and a stoichiometry of 0.15 mol of S-100/mol of polymerized tubulin. Under certain conditions, S-100 proteins also cosediment with MTs prepared by coassembly of S-100 with MTs, probably in the form of an S-100-tubulin complex. Because S-100 binds to MTs under conditions where this protein fraction does not produce observable effects on the kinetics of assembly-disassembly, e.g., in the absence of Ca2+ at pH 6.7, we conclude that the S-100 binding to MTs does not affect the stability of MTs per se, but rather creates conditions for increased sensitivity of MTs to Ca2+. PMID- 2564421 TI - Effect of pyrithiamine treatment and subsequent thiamine rehabilitation on regional cerebral amino acids and thiamine-dependent enzymes. AB - Pyrithiamine-induced thiamine-deficiency encephalopathy in the rat shows many neuropathological and biochemical similarities to Wernicke's encephalopathy in humans. Treatment of rats with pyrithiamine resulted in moderate reductions of glutamate in thalamus and pons and in generalized severe reductions of aspartate in pons (by 89%, p less than 0.01), thalamus (by 83%, p less than 0.01), cerebellum (by 53%, p less than 0.01), and cerebral cortex (by 33%, p less than 0.05). Alanine concentrations were concomitantly increased. Activities of the thiamine-dependent enzyme alpha-ketoglutarate dehydrogenase (alpha KGDH) were decreased in parallel with the aspartate decreases; pyruvate dehydrogenase complex activities were unchanged in all brain regions. Following thiamine administration to symptomatic pyrithiamine-treated rats, neurological symptoms were reversed and concentrations of glutamate, aspartate, and alanine, as well as alpha KGDH activities, were restored to normal in cerebral cortex and pons. Aspartate levels and alpha KGDH activities remained below normal values, however, in thalamus. Thus, pyrithiamine treatment leads to reductions of cerebral alpha KGDH and (1) decreased glucose (pyruvate) oxidation resulting in accumulation of alanine and (2) decreased brain content of glutamate and aspartate. Such changes may be of key significance in the pathophysiology of the reversible and irreversible signs of Wernicke's encephalopathy in humans. PMID- 2564422 TI - Aspartate aminotransferase for synthesis of transmitter glutamate in the medulla oblongata: effect of aminooxyacetic acid and 2-oxoglutarate. AB - The effects of aminooxyacetic acid (AOAA), a transaminase inhibitor, and 2 oxoglutarate, a precursor to glutamate by the activity of aspartate aminotransferase (AAT), on slices of rat medulla oblongata, cerebellum, cerebral cortex, and hippocampus were studied. The slices were superfused and electrically stimulated. There was a Ca2+-dependent stimulus-evoked release of endogenous glutamate, gamma-aminobutyric acid (GABA), and beta-alanine in all regions examined. AOAA (10(-4) and 10(-3) M) decreased the release of glutamate in the medulla oblongata and cerebellum but not in the hippocampus. L-Canaline, a specific inhibitor of ornithine aminotransferase, did not affect the glutamate release in the medulla. 2-Oxoglutarate (10(-3) M) increased the release of glutamate in the medulla oblongata and cerebellum but not in the cerebral cortex and hippocampus. Treatment with AOAA (10(-4) M) almost abolished the activities of AAT in all regions studied. AOAA (10(-4) and 10(-3) M) increased the stimulus evoked release of GABA in the cerebellum, cerebral cortex, and hippocampus, whereas the stimulus-evoked release of beta-alanine was decreased by this agent in all regions studied. These results suggest the participation of AAT in the synthesis of the transmitter glutamate in the medulla oblongata and cerebellum of the rat. PMID- 2564423 TI - Synthesis and release of dopamine in rat brain: comparison between substantia nigra pars compacts, pars reticulata, and striatum. AB - Dopamine (DA) is synthesized and released not only from the terminals of the nigrostriatal dopaminergic neuronal pathway, but also from the dendrites in the substantia nigra. We have investigated the regulation of the DA turnover, the DA synthesis rate, and the DA release in the substantia nigra pars compacts (SNpc) and pars reticulata (SNpr) in vivo. As a measure of DA turnover, we have assessed the concentrations of 3,4-dihydroxyphenylacetic acid and homovanillic acid. As a measure of the DA synthesis rate, we have determined the 3,4 dihydroxyphenylalanine accumulation after inhibition of aromatic L-amino acid decarboxylase by 3-hydroxybenzylhydrazine. As a measure of DA release, we have investigated the disappearance rate of DA after inhibition of its synthesis by alpha-methyl-p-tyrosine and the 3-methoxytyramine accumulation following monoamine oxidase inhibition by pargyline. Both the DA turnover and the DA synthesis rate increased following treatment with the DA receptor antagonist haloperidol and decreased following treatment with the DA receptor agonist apomorphine in the SNpc and in the SNpr, but the effects of the drugs were less pronounced than in the striatum. gamma-Butyrolactone treatment, which suppresses the firing of the dopaminergic neurons, increased the DA synthesis rate in the striatum (165%), but had no such effect in the SNpc or SNpr. Haloperidol, apomorphine, and gamma-butyrolactone increased, decreased, and abolished, respectively, the DA release in the striatum, but the drugs had no or only slight effects on the alpha-methyl-p-tyrosine-induced DA disappearance and on the pargyline-induced 3-methoxytyramine accumulation in the SNpc or SNpr. Taken together, these results indicate that the DA synthesis rate, but not the DA release, are influenced by DA receptor activity and neuronal firing in the SNpc and SNpr. This is in contrast to the situation in the striatum, where both the DA synthesis rate and the DA release are under such control. PMID- 2564424 TI - Endogenous amino acid release from cultured cerebellar neuronal cells: effect of tetanus toxin on glutamate release. AB - Endogenous amino acid release was measured in developing cerebellar neuronal cells in primary culture. In the presence of 25 mM K+ added to the culture medium, cerebellar cells survived more than 3 weeks and showed a high level of differentiation. These cultures are highly enriched in neurons, and electron microscopic observation of these cells after 12 days in vitro (DIV) confirmed the presence of a very large proportion of cells with the morphological characteristics of granule cells, making synapses containing many synaptic vesicles. Synaptogenesis was also confirmed by immunostaining the cells with antisera against synapsin I and synaptophysin, two proteins associated with synaptic vesicles. From these cultures, endogenous glutamate release stimulated by 56 mM K+ was already detected after only a few days in culture, the maximal release value (1,579% increase over basal release) being reached after 10 DIV. In addition to that of glutamate, the release of aspartate, asparagine, alanine, and, particularly, gamma-aminobutyric acid (GABA) was stimulated by 56 mM K+ after 14 DIV, but to a lesser extent. No increase in serine, glutamine, taurine, or tyrosine release was observed during K+ depolarization. The effect of K+ on amino acid release was strictly Ca2+-dependent. Stimulation of the cells with veratridine resulted in a qualitatively similar effect on endogenous amino acid release. In the absence of Ca2+, 30% of the veratridine effect persisted. The Ca2+-dependent release was quantitatively similar after stimulation by veratridine and K+. Treatment of cerebellar cells with tetanus toxin (5 micrograms/ml) for 24 h resulted in a total inhibition of the Ca2+-dependent component of the glutamate release evoked by K+ or veratridine. It is concluded that glutamate is the main amino acid neurotransmitter of cerebellar cells developed in primary culture under the present conditions and that glutamate is probably mainly released through the exocytosis of synaptic vesicles. PMID- 2564425 TI - Purification of two dipeptidyl aminopeptidases II from rat brain and their action on proline-containing neuropeptides. AB - From the soluble and membrane fractions of rat brain homogenate, two enzymes that liberate dipeptides of the type Xaa-Pro from chromogenic substrates were purified to homogeneity. The two isolated dipeptidyl peptidases had similar molecular and catalytic properties: For the native proteins, molecular weights of 110,000 were estimated; for the denatured proteins, the estimate was 52,500. Whereas the soluble peptidase yielded one band of pI 4.2 after analytical isoelectric focusing, two additional enzymatic active bands were detected between pI 4.2 and 4.3 for the membrane-associated form. As judged from identical patterns after neuraminidase treatment, both peptidases contained no sialic acid. A pH optimum of 5.5 was estimated for the hydrolysis of Gly-Pro- and Arg-Pro-nitroanilide. Substrates with alanine instead of proline in the penultimate position were hydrolyzed at comparable rates. Acidic amino acids in the ultimate N-terminal position of the substrates reduced the activities of the peptidases 100-fold as compared with corresponding substrates with unblocked neutral or, especially, basic termini. The action of the dipeptidyl peptidase on several peptides with N terminal Xaa-Pro sequences was investigated. Tripeptides were rapidly hydrolyzed, but the activities considerably decreased with increasing chain length of the peptides. Although the tetrapeptide substance P 1-4 was still a good substrate, the activities detected for the sequential liberation of Xaa-Pro dipeptides from substance P itself or casomorphin were considerably lower. Longer peptides were not cleaved. The peptidases hydrolyzed Pro-Pro bonds, e.g., in bradykinin 1-3 or 1-5 fragments, but bradykinin itself was resistant. The enzymes were inhibited by serine protease inhibitors, like diisopropyl fluorophosphate or phenylmethylsulfonyl fluoride, and by high salt concentrations but not by the aminopeptidase inhibitors bacitracin and bestatin. Based on the molecular and catalytic properties, both enzymes can be classified as species of dipeptidyl peptidase II (EC 3.4.14.2) rather than IV (EC 3.4.14.5). However, some catalytic properties differentiate the brain enzyme from forms of dipeptidyl peptidase II of other sources. PMID- 2564426 TI - Glutamate stimulation of [3H]dopamine release from dissociated cell cultures of rat ventral mesencephalon. AB - In dissociated cell cultures of fetal rat ventral mesencephalon preloaded with [3H]dopamine, glutamate (10(-5)-10(-3) M) stimulated the release of [3H]dopamine. Glutamate stimulation of [3H]dopamine release was Ca2+ dependent and was blocked by the glutamate antagonist, cis-2,3-piperidine dicarboxylic acid. Glutamate stimulation of [3H]dopamine release was not due to glutamate neurotoxicity because (1) glutamate did not cause release of a cytosolic marker, lactate dehydrogenase, and (2) preincubation of cultures with glutamate did not impair subsequent ability of the cells to take up or release [3H]dopamine. Thus, these dissociated cell cultures appear to provide a good model system to characterize glutamate stimulation of dopamine release. Release of [3H]dopamine from these cultures was stimulated by veratridine, an activator of voltage-sensitive Na+ channels, and this stimulation was blocked by tetrodotoxin. However, glutamate stimulated [3H]dopamine release was not blocked by tetrodotoxin or Zn2+. Substitution of NaCl in the extracellular medium by sucrose, LiCl, or Na2SO4 had no effect on glutamate stimulation of [3H]dopamine release; however, release was inhibited when NaCl was replaced by choline chloride or N-methyl-D-glucamine HCl. Glutamate-stimulated [3H]-dopamine release was well maintained (60-82% of control) in the presence of Co2+, which blocks Ca2+ action potentials, and was unaffected by the local anesthetic, lidocaine. These results are discussed in terms of the receptor and ionic mechanisms involved in the stimulation of dopamine release by excitatory amino acids. PMID- 2564427 TI - Clinical trials of agents that reverse multidrug-resistance. PMID- 2564428 TI - Drug-resistance in multiple myeloma and non-Hodgkin's lymphoma: detection of P glycoprotein and potential circumvention by addition of verapamil to chemotherapy. AB - The B-cell neoplasms, multiple myeloma and non-Hodgkin's lymphoma, frequently become drug resistant, despite initial responses to chemotherapeutic drugs. Tumor cells from eight patients with clinically drug-refractory disease were evaluated by immuno-histochemical staining for monoclonal immunoglobulin (Ig) expression, nuclear proliferation antigen, P-glycoprotein (P-gly) expression, and other cellular antigens. P-gly was detected on tumor cells from six of eight patients with drug-resistant disease. Of the six patients with P-gly-positive tumors, five patients had advanced multiple myeloma and one had a drug-refractory non Hodgkin's lymphoma. Cellular RNA analysis confirmed the over-expression of P-gly. In an effort to overcome drug resistance, a pilot study evaluated possible verapamil enhancement of chemotherapy in these eight patients. All patients had developed progressive disease while receiving a regimen containing vincristine and doxorubicin, and seven of eight patients had previously received continuous infusion vincristine and doxorubicin plus oral dexamethasone (VAD). At the time of progressive disease, continuous infusion verapamil was added to the VAD regimen. Three of the eight patients who were refractory to vincristine and doxorubicin alone responded when verapamil was added to VAD. The three patients who responded had P-gly-positive tumors. Verapamil increased the intracellular accumulation of doxorubicin and vincristine in vitro for both a P-gly-positive myeloma cell line and tumor cells from two patients with end-stage myeloma which over-expressed P-gly. The dose-limiting side effect associated with the addition of verapamil to chemotherapy was temporary impairment of cardiac function, manifest as hypotension and cardiac arrhythmia. We conclude that P-gly expression occurs in drug-refractory B-cell neoplasms and may contribute to the development of clinical drug resistance. However, other factors, such as the proliferative activity of the tumor, may also play a role in determining response to chemotherapy. The administration of verapamil along with VAD chemotherapy may partially circumvent drug resistance in patients whose tumors over-express P-gly. PMID- 2564429 TI - A muscle acetylcholine receptor is expressed in the human cerebellar medulloblastoma cell line TE671. AB - The human neuromedulloblastoma cell line TE671 is shown by single-channel recordings to express nicotinic acetylcholine receptors (AChRs) that are blocked by alpha-bungarotoxin (alpha Bgt). These AChRs do not react with antisera to the alpha Bgt-binding protein of brain or with monoclonal antibodies (mAbs) to brain nicotinic AChRs that do not bind alpha Bgt. TE671 AChRs do react with autoantibodies to muscle AChRs from myasthenia gravis patients and with mAbs to muscle AChRs, including mAbs specific for extrajunctional AChRs. AChRs. AChRs purified from TE671 cells are composed of 4 kinds of subunits corresponding to those of muscle AChR. Sequences of cDNAs for the ACh-binding alpha subunit and the delta subunit of this AChR further identify it as muscle AChR. Expression of TE671 AChR can be up-regulated by nicotine and dexamethasone, and down-regulated by forskolin. PMID- 2564430 TI - Vasopressin neuromodulation in the hippocampus. AB - This study explored an effector mechanism associated with the arginine vasopressin (AVP) recognition site in the hippocampus, namely, potentiation of norepinephrine (NE)-induced cAMP accumulation in the surviving hippocampal slice. The biochemical mechanisms that underlie the AVP potentiation were investigated as follows: First, the actions of AVP upon NE-induced accumulation of cAMP in hippocampal slices from rat brain were specific to AVP and not shared by other closely related peptides, namely, oxytocin and AVP4-9. Second, the AVP-induced neuromodulation involved beta-adrenergic receptors, with AVP having no effect on cAMP levels in the absence of NE. Third, the potentiation by AVP was biphasic, with lower AVP concentrations potentiating NE-induced cAMP accumulation, while higher concentrations did not potentiate. Fourth, an antagonist of V1-type AVP receptors blocked AVP potentiation. Fifth, AVP potentiation was dependent upon extracellular calcium concentrations. Sixth, AVP potentiation was blocked by 50 microM trifluoperazine, which is consistent with a calcium-calmodulin involvement but which might also implicate protein kinase C. These alternatives and the nature of the calcium involvement are discussed. AVP actions thus appear to involve interactions between several second-messenger systems and suggest a biochemical mechanism by which AVP exerts its centrally mediated behavioral effects. PMID- 2564431 TI - Release of glutamate and of free fatty acids in vasogenic brain edema. AB - The pathophysiological potential of mediator substances in manifestations of secondary brain damage is attracting increased attention. This is particularly true of the excitatory transmitters glutamate and arachidonic acid. Noxious properties of these compounds in central nervous tissue have been demonstrated. The current study was performed to determine whether glutamate and arachidonate are released in brain tissue secondary to focal trauma. For this purpose, a cold injury of exposed cerebral cortex was induced in cats. Marked accumulation of glutamate was observed in interstitially drained edema fluid, reaching 10 to 15 times the level that was assessed in normal cerebrospinal fluid (CSF) prior to trauma. The extracellular release of glutamate was further dramatically enhanced by a critical decrease of the cerebral perfusion pressure due to a malignant increase of intracranial pressure. Under these conditions, glutamate concentrations 1000 to 1500 times normal levels accumulated in vasogenic edema fluid, demonstrating a relationship between the extent of the release of glutamate in damaged brain and the severity of the insult. Although under normal conditions glutamate concentrations in plasma were considerably higher than in the interstitial fluid, the pronounced increase of glutamate in this compartment due to trauma cannot be explained by transport of the compound together with the plasma-like edema from the intravascular space. Corresponding findings were obtained for free fatty acid concentrations in edema fluid. Almost all fatty acids that were studied had a significantly higher concentration in edema fluid than in normal CSF obtained as a control prior to trauma. However, contrary to the findings for glutamate, fatty acid concentrations in edema fluid were lower than in plasma. Accumulation of fatty acids in vasogenic edema fluid might, therefore, have resulted from uptake of the material together with edema fluid through the breached blood-brain barrier. Arachidonic acid was an exception. Its concentrations were significantly higher in edema fluid than in plasma, suggesting that it was released from cerebral parenchyma as the underlying mechanism of its extracellular accumulation. The current observations provide further support for a mediator function of glutamate and arachidonic acid in acute traumatic lesions of the brain. Quantitative assessment of the release of highly active mediator substances in brain tissue may facilitate analysis of the therapeutic efficiency of specific treatment aimed at interfering with the release or pathological function of mediators of secondary brain damage. PMID- 2564432 TI - Prenatal diagnosis and heterozygote detection by DNA analysis in ornithine transcarbamylase deficiency. AB - This report summarizes our experience with DNA analysis using a complementary DNA probe for ornithine transcarbamylase in 24 individuals or families with deficiency of this enzyme. In four cases, including three reported elsewhere, a Taql restriction site alteration directly detected the mutation. In 10 additional cases, only an affected male was available, and results of DNA analysis using the Taql enzyme were normal. In 10 cases, family studies were performed with the use of restriction fragment length polymorphisms. Prenatal diagnostic studies were performed for three informative pregnancies, and two affected male fetuses were identified. Analysis of two restriction fragment length polymorphisms, Mspla and BamHl, was informative in 14 of 19 (74%) known carrier females and in 21 of 35 (60%) females (the total number studied). One female previously predicted to be a noncarrier by protein-loading test was determined to be a carrier by analysis of restriction fragment length polymorphisms. The frequency of Taql site alterations was 4 of 24 families (17%). These data illustrate the importance of DNA analysis, pedigree analysis, and biochemical testing in families with ornithine transcarbamylase deficiency to detect carriers and establish the diagnosis prenatally. PMID- 2564433 TI - Genetic counseling for phenylketonuria. PMID- 2564434 TI - Histogenesis of dieldrin and DDT-induced hepatocellular carcinoma in Balb/c mice. AB - Male, Balb/c mice were fed diets containing dieldrin (10 ppm) and DDT (100-175 ppm) for 75 weeks. Control and treated mice were serially killed and their livers analyzed by histological and histochemical procedures after 2, 4, 8, 16, 36, 52 and 75 weeks of exposure. Mice administered both chlorinated hydrocarbons initially responded with centrolobular hepatocytomegaly. The cells were characterized by decreased glucose-6-phosphatase and succinate dehydrogenase activity. At later periods 52 through 75 weeks, foci of phenotypically-altered hepatocytes were noted. The cells of these lesions were basophilic or clear staining in hematoxylin and eosin-stained sections and displayed increased gamma glutamyl transpeptidase activity. In mice preloaded with iron dextran, cells of foci were negative for iron when the surrounding parenchyma was siderotic. Hepatocellular adenomas (HA) and carcinomas (HPC) were composed of cells with increased gamma glutamyl transpeptidase and glucose-6-phosphate dehydrogenase and decreased glucose-6-phosphatase and succinate dehydrogenase activity. In iron loaded mice, the cells of HA and HPC did not stain for iron in otherwise siderotic surroundings. Both hepatocellular foci and adenomas may be potential precursors of mouse hepatocellular carcinomas. PMID- 2564435 TI - Neuroleptic-resistant schizophrenics. AB - According to Crow's postulated positive-negative distinction, negative symptoms of schizophrenia are less responsive to neuroleptic drugs. Not all research evidence supports that expectation, however, so that neuroleptics need not be withheld from any schizophrenic patients. Other aspects of Crow's hypothesized distinction have indicated possible promising results, but more research is required. PMID- 2564436 TI - Neuroleptic-induced parkinsonian side effects in the mentally handicapped. AB - Sixty-seven neuroleptic-mediated mentally handicapped subjects in a hospital were studied to determine the prevalence of Parkinsonian side effects. A Parkinsonism scale was devised and administered. Sixty-one per cent of the sample had mild to moderately severe side effects. Sex, age, cumulative and current chlorpromazine doses, cumulative and current anticholinergic doses and anti-epileptic medication status did not predict the Parkinsonism scores. Overt brain damage was not a predictor. The difference between the neuroleptic medicated group and neuroleptic free matched controls was highly significant indicating that the Parkinsonian type of movement disorder was related to neuroleptic medication. PMID- 2564437 TI - Primate evolution of a human chromosome 1 hypervariable repetitive element. AB - The clone designated hMF #1 represents a clustered DNA family, located on chromosome 1, consisting of tandem arrays displaying a monomeric length of 40 bp and a repetition frequency of approximately 7 x 10(3) copies per haploid genome. The sequence hMF #1 reveals multiple restriction fragment length polymorphisms (RFLPs) when human genomic DNA is digested with a variety of 4-6-bp recognition sequence restriction enzymes (i.e., Taq I, Eco RI, Pst I, etc.). When hamster and mouse genomic DNA was digested and analyzed, no cross-species homology could be observed. Further investigation revealed considerable hybridization in the higher primates (chimpanzee, gorilla, and orangutan) as well as some monkey species. The evolutionary relationship of this repetitive DNA sequence, found in humans, to that of other primates was explored using two hybridization methods: DNA dot blot to establish copy number and Southern DNA analysis to examine the complexity of the RFLPs. Homology to the hMF #1 sequence was found throughout the suborder Anthropoidea in 14 ape and New and Old World monkey species. However the sequence was absent in one species of the suborder Prosimii. Several discrepancies between "established" evolutionary relationships and those predicted by hMF #1 exist, which suggests that repetitive elements of this type are not reliable indicators of phylogenetic branching patterns. The phenomenon of marked diversity between sequence homologies and copy numbers of dispersed repetitive DNA of closely related species has been observed in Drosophila, mice, Galago, and higher primates. We report here a similar phenomenon for a clustered repeat that may have originated at an early stage of primate evolution. PMID- 2564438 TI - Evidence of mitochondrial involvement in scrapie infection. AB - Two cDNA libraries were constructed from brain membrane and cytoskeletal preparations purified from scrapie-infected hamster brains. Four recombinants strongly preferential to the scrapie cytoskeletal preparation were identified by the differential hybridization of 7,000 recombinants. These clones were not, however, preferential to total nucleic acids extracted from scrapie-infected hamster brains. DNA sequence analysis revealed all four clones to have significant sequence similarities to the mouse mitochondrial genome. This correlation led us to consider a mitochondrial association with scrapie infectivity. Brain mitochondria were purified by sucrose gradient density centrifugation and found to contain high infectivity. Removal of mitochondrial outer membranes by osmotic shock or digitonin treatment resulted in no detectable loss of titer. PMID- 2564439 TI - Genetic analysis of endogenous xenotropic murine leukemia viruses: association with two common mouse mutations and the viral restriction locus Fv-1. AB - We have defined 40 endogenous xenotropic virus (Xmv) loci from several common inbred strains of mice by examining provirus-cell DNA junction fragments in recombinant inbred mice. Some inbred strains carried unique proviruses, but most Xmv loci were present in several strains, indicating that many Xmv integration events preexisted modern inbreeding. It was also clear that most Xmv junction fragment variation between inbred strains resulted from independent integration events and not modification or restriction site polymorphism following integration. Chromosomal assignments were determined for 32 Xmv loci by comparing their recombinant inbred strain distribution patterns to those of known genetic markers. The Xmv loci were generally dispersed throughout the genome, but several chromosomal regions contained more than one provirus. Furthermore, several close genetic associations with cellular genes were discovered. Four Xmv loci were closely linked to Fv-1b, a dominant viral resistance gene present in C57BL/6J, BALB/cJ, A/J, and several other strains. Xmv-28 was closely linked to rd (retinal degeneration), and Xmv-10 was closely linked to a (non-agouti), both of which are old mutations as inferred from their broad distribution in mice. We suggest that Xmv integration contributed to genetic diversity in the past and that much of this diversity exists today in common laboratory strains. PMID- 2564440 TI - Induction of nephroblastoma by myeloblastosis-associated virus type 1: state of proviral DNAs in tumor cells. AB - Myeloblastosis-associated virus type 1 (MAV1) derived from a molecular clone of infectious proviral DNA (B. Perbal, J. S. Lipsick, J. Svoboda, R. F. Silva, and M. A. Baluda, J. Virol. 56:240-244, 1985) was shown to specifically induce nephroblastoma in chickens and therefore belongs to the MAV-N class. We show that nephroblastomas are polyclonal tumors containing rearranged proviral genomes. Rearrangements occur preferentially in the gag-pol region of the MAV1 proviral genome, and similar rearrangements can be detected in well-developed independent tumors. PMID- 2564441 TI - Human Sertoli-spermatogenic cell cocultures prepared from biopsies of cryptorchid testes performed during orchidopexy. AB - A procedure is described for the preparation and maintenance of human Sertoli spermatogenic cell cocultures using biopsies of normal and undescended testis. The evaluation of cell viability and differentiation potential of cultured spermatogenic cell was monitored by [3H]thymidine labeling combined with light microscopic autoradiography. Spermatogenic cells of the same progeny, connected by intercellular bridges, display synchronous DNA synthesis when labeled at the preleptotene stage of meiotic prophase. The pattern of [35S]methionine-labeled secretory proteins was determined by two-dimensional electrophoresis and autoradiography during testicular development and compared with these observed in human Sertoli-spermatogenic cell cocultures prepared from same specimens. Both testicular tissue and cocultured Sertoli and spermatogenic cells displayed comparable patterns of secretory proteins. A discrete group of acidic polypeptides of Sertoli cell origin enhanced their radiolabeling intensity during testicular development. Results of this paper indicate that human Sertoli spermatogenic cell cocultures could be valuable for assessing the proliferation and differentiation potential of spermatogenic cells in children with cryptorchid testis. PMID- 2564442 TI - Lower urinary tract function in the sheep fetus: studies of autonomic control and pharmacologic responses of the fetal bladder. AB - An experimental preparation was developed to study fetal bladder function. In 16 fetal sheep at 120 days' gestation, two distinct types of bladder contractions were identified, one typical of contractions seen after birth and one phasic. Bladder capacity and intravesical pressure during contraction varied widely among fetuses, but residual urine was invariably less than one ml. Several drugs altered fetal bladder function: bethanechol decreased bladder capacity (6.8 vs. 13.6 ml.); atropine increased bladder capacity (27.3 vs. 6.8 ml.) and resulted in a 17-ml. residual urine; and ritodrine, magnesium sulfate and diltiazem inhibited bladder contractions and also resulted in residual urines of seven, six and seven ml. respectively. The fetal sheep bladder at 120 days' gestation is under both cholinergic and beta-adrenergic control. Further, drugs currently administered to pregnant women for the treatment of pre-term labor and pre-eclampsia cross the placenta and can directly alter bladder function in the developing fetus and the neonate. PMID- 2564443 TI - Residual urine in children with acute cystitis and in healthy children: assessment by sonography. AB - Residual urine was assessed by ultrasound in children with single attacks of symptomatic lower urinary tract infection and in healthy controls. Residual urine was found significantly more often in the 39 patients during acute illness as well as during a follow-up of 6 months, compared to 55 control children. Infections caused by P-fimbriated Escherichia coli were not more often associated with residual urine than infections with nonP-fimbriated Escherichia coli or other bacterial species. It is suggested that residual urine is a facilitating host factor among others in the pathogenesis of symptomatic urinary tract infection in childhood. PMID- 2564444 TI - Ureteral obstruction caused by vasculitis. AB - We report 2 cases of ureteral obstruction caused by vasculitis (polyarteritis nodosa and systemic lupus erythematosus). Case 1, in which the diagnosis was unknown preoperatively, was managed surgically with ureteral resection and reimplantation. Case 2 was managed medically with chlorambucil and corticosteroids, which resulted in resolution of ureteral obstruction. PMID- 2564445 TI - Beta-blockers and primary prevention of coronary heart disease in patients with high blood pressure. AB - We conducted a population-based, case-control study to determine whether beta blockers, used for the treatment of hypertension, prevent first events of coronary heart disease. Cases were patients who had high blood pressure treated with medicines and who presented in 1982 to 1984 with angina or fatal or nonfatal myocardial infarction. Controls were a probability sample of health maintenance organization patients with pharmacologically treated hypertension and free of coronary heart disease. Blinded to case-control status, we reviewed the medical records of the 248 cases and 737 controls. The health maintenance organization's computerized pharmacy database was used to ascertain the use of beta-blockers. Fewer cases than controls were taking beta-blockers. This difference was confined to those with nonfatal infarctions. After adjustment for confounding, the estimated relative risk was 0.62 (95% confidence interval, 0.39 to 0.99). Higher doses of beta-blockers conferred greater protection. We conclude that beta blockers may prevent first events of nonfatal myocardial infarction in patients with high blood pressure. PMID- 2564446 TI - Immunohistochemical demonstration of somatostatin-containing cells in the equine thyroid and parathyroid glands. PMID- 2564447 TI - [Continuing education within the framework of the annual meeting of the European Nursing Students Group 1988 in Berne]. PMID- 2564448 TI - [Importance of inhalation therapy in bronchial diseases]. PMID- 2564449 TI - Surface and cytoplasmic expression of CD2, CD13, and CD25 antigens in human T lymphotropic virus type I infected cell line. An immunoelectron microscopic study. AB - Immunoperoxidase electron microscopy revealed intracellular expression of CD2, CD13, and CD25 antigens in a unique human T lymphotropic virus type I infected cell line, TI-CL. This cell line is thought to be derived from cord mononuclear cells in the bifurcation stage of T cell and myelomonocytic lineage, based on cytochemical, immunologic, and molecular genetic analyses. Each antigen expression was observed on the plasma membranes, the cisternae of rough endoplasmic reticulum, and the perinuclear cisternae. Surface and cytoplasmic expression of CD25 antigen in adult T cell leukemia/lymphoma cells have been reported, but similar events concerning CD2 and CD13 antigens have not been described previously. CD13 antigen is a marker of myelomonocytic lineage, and biochemical analyses have demonstrated that monoclonal antibodies belonging to CD13 precipitated two glycoprotein molecules of 130,000 and 150,000 molecular weight and that the former molecule (gp130) is the precursor protein of the latter (gp150). In our findings, CD13-positive reaction in the cisternae of rough endoplasmic reticulum and the perinuclear cisternae may correspond to the precursor protein of CD13. CD2 antigen, a 50 kilodalton sheep erythrocyte rosette receptor protein, is the early T lineage antigen and plays an important role in T cell activation. The detection of CD2 antigen on the cell surface and in the cytoplasm may morphologically confirm that this antigen is synthesized in the cytoplasm and inserted into the plasma membrane. PMID- 2564450 TI - Evidence for the in vivo production of interleukin-3 in lethally irradiated mice transplanted with syngeneic murine splenocytes. AB - We wish to demonstrate new evidence for the in vivo production of interleukin-3 (IL-3). Syngeneic murine splenocytes were transplanted into lethally irradiated mice. The spleen cells in these transplant mice spontaneously produced IL-3 in cultures, and IL-3-like activity was detected in the serum. Chronological changes of the Thy-1 antigen expression on the bone marrow cells, and the number of myeloid progenitors in the bone marrow from post-transplant mice correlated with the amount of IL-3 produced in vivo. These results suggest that IL-3 may be produced by activated T cells during the syngeneic mixed lymphocyte reaction induced in vivo. PMID- 2564451 TI - IMP dehydrogenase: inhibition by the anti-leukemic drug, tiazofurin. AB - Tiazofurin through its active metabolite thiazole-4-carboxamide adenine dinucleotide (TAD) inhibits IMP dehydrogenase, the rate-limiting enzyme of GTP biosynthesis. IMP dehydrogenase activity in human leukemic cell extracts (33.4 +/ 0.1 nmol/h/mg protein) was increased 11-fold compared to normal leukocytes (3.1 +/- 0.5). Km values for IMP and NAD+ of leukemic IMP dehydrogenase were 22.7 and 44.0 microM, respectively. XMP inhibited competitively with IMP and noncompetitively with NAD+. NADH exerted mixed type inhibition with respect to both IMP and NAD+. The inhibitory pattern of TAD was quite similar to that of NADH; however, the affinity of TAD to leukemic IMP dehydrogenase (Ki = 0.1 microM) was three orders of magnitude higher than the natural product NADH (Ki = 150 microM). These results contribute to an understanding of the mechanism of action of tiazofurin in the treatment of leukemia. PMID- 2564452 TI - Acute myeloid leukemia: analysis of ras gene mutations and clonality defined by polymorphic X-linked loci. AB - In vitro DNA amplification and synthetic oligonucleotide hybridization was used to analyze 57 acute myelocytic leukemias (AML) for the presence of ras gene mutations. We demonstrated mutated alleles in 19% of primary AMLs (10/51) as well as in five of six secondary leukemias. Mutations occurred predominantly at N-ras codons 12, 13, or 61 (13 cases) and twice at Ki-ras codons 12 and 13. Ras gene mutations were preferentially associated with an M4 morphology according to the FAB (French-American-British) classification, but no particular correlation was observed with respect to clinical parameter (sex, age, course of disease) or immunophenotype and karyotype. Mutated ras alleles were absent in nine mutation positive cases analyzed during remission. However, a more complex pattern emerged from the five patients analyzed in relapse exhibiting identical ras mutations in three cases, absence of a mutated allele in one patient, and acquisition of a N ras mutation in yet another case, in which no mutation had been detected initially. Moreover, restriction fragment length polymorphisms (RFLP) of the X chromosome genes hypoxanthine phosphoribosyl transferase (HPRT) and phosphoglycerate kinase (PGK) were studied in 19 of the AML patients. Nine cases (47%) were heterozygous for BglI or BamHI RFLPs at the PGK or HPRT loci, respectively, and therefore suitable for clonal analysis investigating X chromosome inactivation. All of the patients exhibited a monoclonal leukemic cell population at presentation. In addition, five of seven cases studied in remission showed reemergence of a polyclonal pattern. However, two children exhibited persistence of monoclonal hematopoiesis despite complete clinical/hematological remission and a corresponding loss of a mutated ras allele in one of the patients. These data indicate the value of molecular genetic approaches for evaluation of the heterogeneous nature of remission and relapse in AML. PMID- 2564453 TI - Biochemical characterization of U937 cells resistant to L-asparaginase: the role of asparagine synthetase. AB - A human histiocytic lymphoma cell line, U937, is highly sensitive to L asparaginase with an ID50 of about 0.0001 U/ml after 72 hr of culture. When U937 cells were made resistant to either L-asparaginase (1 U/ml) or asparagine deprivation, the activity of asparagine synthetase increased to 80- or 7-fold of the wild type, respectively. The phenotype of the resistance to L-asparaginase turned out to be stable under nonselective conditions for over several months. The hybrids between L-asparaginase sensitive (Molt4) and resistant (HL-60) cell lines revealed the latter phenotype in terms of L-asparaginase sensitivity and the activity of asparagine synthetase. Furthermore, U937 cells resistant to L asparaginase could survive in glutamine-free media with 1.5-fold elevation of glutamine synthetase activity. These results altogether clarify the role of asparagine synthetase in L-asparaginase toxicity and have a good implication for the clinical use of L-asparaginase. PMID- 2564454 TI - [In which patients is beta blockade in myocardial infarction most effective?]. PMID- 2564455 TI - [Medical ethics. A symposium of the Swedish Medical Society in 1988. Ethics and reproduction technology]. PMID- 2564456 TI - Increased risk of breast cancer after low-dose irradiation. AB - A significant increase in the risk of breast cancer has been found for the most recent 5-year period of a long-term follow-up study of children subjected to scalp irradiation, in whom a carcinogenic effect was previously only apparent in the head and neck. This increased risk was found among women aged 5 to 9 years at exposure. The breast had been exposed to a low radiation dose of approximately 16 mGy. PMID- 2564457 TI - Detection of polymorphisms at cytosine phosphoguanadine dinucleotides and diagnosis of haemophilia B carriers. AB - The polymerase chain reaction procedure (PCR) was used to detect a polymorphic Hha I site adjacent to the factor IX locus in a panel of 33 phenotypically normal caucasian individuals. This technique was also applied to a haemophilia B family pedigree. The Hha I polymorphic site was located 8 kb 3' to the factor IX gene, and the proportion of female subjects expected to be heterozygous at this site was 0.48. The Hha I locus was in linkage equilibrium with the other polymorphic loci on the factor IX gene. These findings, besides increasing the proportion of caucasian individuals whose haemophilia B carrier state can be diagnosed from 79% to 89%, demonstrate this widely applicable use of PCR for the detection of DNA polymorphism at cytosine phosphoguanadine dinucleotides irrespective of the methylation status. PMID- 2564458 TI - Long-term interferon therapy for thrombocytosis in myeloproliferative diseases. AB - 31 patients with thrombocytosis associated with myeloproliferative disorders were included in a prospective trial of long-term interferon therapy. 6 patients (19%) had side-effects which required withdrawal of interferon within one year. 22 patients (71%) achieved and maintained a complete response (platelet count less than 440 x 10(9)/l) for at least twelve months, with reduction or abolition of symptoms associated with thrombocytosis and a significant fall in bone-marrow megakaryocytes. At twelve months, 25 patients were randomly allocated to maintenance or withdrawal of interferon. Thrombocytosis recurred rapidly when treatment was stopped, but a second remission could be achieved by resumption of interferon therapy. PMID- 2564459 TI - Decline in skeletal muscle mitochondrial respiratory chain function: possible factor in ageing. AB - State III (activated) mitochondrial respiration rates with pyruvate/malate, glutamate/malate, and succinate as substrates were assayed in isolated intact skeletal muscle mitochondria in 29 subjects aged 16-92 years. There was a significant negative correlation between respiration rate and age with all substrates tested. A similar trend was seen for respiratory enzyme activities assayed in muscle homogenate. These findings suggest a substantial fall in mitochondrial oxidative capacity in ageing muscle, which may contribute to reduced exercise capacity in elderly people. Mitochondrial respiratory failure may contribute to the ageing process in other organs. PMID- 2564460 TI - Lignocaine metabolite formation as a measure of pre-transplant liver function. AB - A method for rapid assessment of hepatic function in liver donors based on the formation of the lignocaine metabolite monoethylglycinexylidide (MEGX), was used in a prospective study of 69 donor-recipient pairs. The probability of graft survival over 120 days was significantly higher for livers from donors with MEGX test values above 90 micrograms/l than for those from donors with MEGX values of 90 micrograms/l or below. Other liver function tests (bilirubin, prothrombin time, activity of aminotransferases, glutamate dehydrogenase, and cholinesterase, indocyanine green clearance, and galactose elimination capacity) were inefficient at predicting early outcome of transplantation. For a 20-day graft survival, the MEGX test showed prognostic sensitivity of 73% and specificity of 78%. These findings suggest that the MEGX formation test could be valuable for selection of donor organs. PMID- 2564461 TI - Mitochondrial DNA mutations as an important contributor to ageing and degenerative diseases. AB - The human mitochondrial genome is very small and economically packed; the expression of the whole genome is essential for the maintenance of mitochondrial bioenergetic function. Mutation occurs at a much higher rate in the mitochondrial DNA (mtDNA) than in chromosomal DNA. Transient heteroplasmy of mtDNA occurs after a mutational event; the random pattern of cytoplasmic segregation that occurs during subsequent growth gives rise to a mosaic of cells. The variable proportion of mutant mitochondrial genomes per cell results in cells with a range of bioenergetic capacities. It is proposed that the accumulation of mitochondrial mutations and the subsequent cytoplasmic segregation of these mutations during life is an important contributor both to the ageing process and to several human degenerative diseases. Replacement therapy and pharmacological support may be possible for the amelioration of such disorders by means of appropriate redox compounds. Moreover, new compounds with desired redox potentials can be rationally designed for clinical use. PMID- 2564462 TI - Meningococcal meningitis. PMID- 2564463 TI - Self-tolerance and autoimmunity: bridging the gap. PMID- 2564464 TI - Outcome of depression. PMID- 2564465 TI - Nausea and vomiting after general anaesthesia. PMID- 2564466 TI - Transdermal testosterone. PMID- 2564467 TI - Training of counsellors on sickle-cell disorders in Africa. AB - Falling infant and childhood mortality rates, especially in urban centres, have allowed greater survival of individuals with sickle-cell disorders (SCD), and the need to provide appropriate services has become pressing. The training and employment of counsellors on SCD, shown here to be popular and feasible, seems an essential first step towards the development of a community-based and appropriate policy for coping with SCD in Africa. PMID- 2564468 TI - Prevention of Pneumocystis carinii pneumonia by inhaled pentamidine. AB - The efficacy and toxicity of pentamidine inhaled once a month to prevent Pneumocystis carinii pneumonia (PCP) was investigated in 102 patients with the acquired immunodeficiency syndrome (AIDS) or AIDS-related complex (ARC). The cohort was compared with historical controls after a mean duration of prophylaxis of 6.38 months. 86% and 15% of the patients had AIDS or ARC, respectively. 50% of patients had had one previous episode of PCP, 9% had had two episodes, and 3% had had three. 11 patients acquired PCP. Among these 51 patients with one prior episode of PCP, the PCP-free survival after 3.03, 4.7, and 6.38 months of prophylaxis was 98%, 92%, and 82%, respectively. Compared with those for historical controls, the data suggest that inhaled pentamidine can delay relapse by 6 months and reduce the rate of relapse by 50%. PCP acquired while patients were inhaling pentamidine prophylactically was mild and had a case-fatality rate of only 9%. Further investigation of the prophylactic value of inhaled pentamidine is warranted. PMID- 2564469 TI - Comfrey herb tea and hepatic veno-occlusive disease. PMID- 2564470 TI - Tilting towards a diagnosis in recurrent unexplained syncope. AB - 71 patients with recurrent syncope which remained unexplained after standard clinical and electrophysiological investigation underwent 60 degree head-up tilt. This procedure reproduced symptoms with vasovagal syncope in 53 (74%), 40 of whom had bradycardia, some with prolonged asystole, during syncope. The other 13 patients had predominant vasodepression with hypotension. Mean time to syncope after tilt was 25 min. Patients with conduction tissue disease and age-matched control subjects had a 15% and 7% incidence of tilt syncope, respectively. Temporary dual-chamber pacing aborted syncope in 85% of subjects, and improved cardiac index and systemic blood pressure during tilt. Long-term results indicate that selected patients may benefit from permanent dual-chamber pacing. Head-up tilt is useful in the investigation of unexplained syncope because symptoms are reproduced in front of a medical witness. PMID- 2564471 TI - Called to account. Obstetrics. Pediatrics. PMID- 2564472 TI - Desmopressin and myocardial infarction. PMID- 2564473 TI - Hepatitis delta infection in north-west Spain. PMID- 2564474 TI - Inhaled budesonide for acute wheeze in infants. PMID- 2564475 TI - 99mTc-HMPAO washout from brain. PMID- 2564476 TI - Totally implantable vascular access for antimicrobial infusion at home and prevention of systemic candidosis. PMID- 2564477 TI - Hypoxic-reperfusion injury in inflamed joints. PMID- 2564478 TI - Osseous malformation in baby born to woman on cyclosporin. PMID- 2564479 TI - Cutaneous alternariosis responding to ketoconazole. PMID- 2564480 TI - Altered c-H-ras gene in non-neoplastic tissue. PMID- 2564481 TI - Duchenne muscular dystrophy: neonatal screening and prenatal diagnosis. PMID- 2564482 TI - Warfarin to prevent thromboembolism in chronic atrial fibrillation. PMID- 2564483 TI - Antineutrophil cytoplasmic antibodies. PMID- 2564484 TI - Amiodarone prophylaxis in hypertrophic obstructive cardiomyopathy. PMID- 2564485 TI - Measuring serum gonadotropins: a cautionary note. PMID- 2564486 TI - Visually read HIV immunoassays. PMID- 2564487 TI - Reduced CD4 + T cells and severe oral candidiasis in absence of HIV infection. PMID- 2564488 TI - Occupational asthma in a chicory grower. PMID- 2564489 TI - Clinical signs of pneumonia in children. PMID- 2564490 TI - Trends in unexpected infant deaths. PMID- 2564491 TI - Myocardial injury after interleukin-2 therapy. PMID- 2564492 TI - Nephrotic syndrome: from toddlers to twenties. PMID- 2564493 TI - Pelvic floor support in constipation. PMID- 2564494 TI - Acute hepatic failure in hepatitis A. PMID- 2564495 TI - Subcutaneous apomorphine as a diagnostic test for dopaminergic responsiveness in parkinsonian syndromes. PMID- 2564496 TI - Increasing number of organ donors. PMID- 2564497 TI - Israel and The Lancet. PMID- 2564498 TI - Safe motherhood: the Niamey conference. PMID- 2564499 TI - Occupations, deaths, and coroners' inquiries. PMID- 2564500 TI - Pancreatic transplantation. PMID- 2564501 TI - Laterality of breast cancer associated with repeated chest fluoroscopies. PMID- 2564502 TI - Chorionic villus sampling versus amniocentesis. PMID- 2564503 TI - Ultrasound screening for fetal malformations. PMID- 2564504 TI - AIDS in the UK and world wide. PMID- 2564505 TI - Effect of theophylline on exercise-induced myocardial ischaemia. AB - In a single-blind, placebo-controlled, randomised trial in 20 patients with stable angina pectoris, intravenous theophylline ethylenediamine (aminophylline), 7 mg/kg, increased the time to onset of angina by 46%, the heart-rate/blood pressure product (an index of myocardial oxygen consumption) at 1 mm ST segment depression by 22%, and exercise duration by 24%. In a subsequent double-blind placebo-controlled trial in 8 patients a single oral dose of theophylline (375 mg) increased the time to onset of angina by 56%, the heart-rate/blood-pressure product at 1 mm ST segment depression by 22%, and the exercise duration by 35%. Infusion of theophylline ethylenediamine during angiography (10 patients) did not affect the diameter of epicardial coronary arteries. The beneficial effects of theophylline may be due to redistribution of coronary blood flow from non ischaemic to ischaemic myocardium. PMID- 2564506 TI - Long-term use of thiazide diuretics and risk of hip fracture. AB - To assess whether long-term thiazide use is associated with a decreased risk of hip fracture, a nested case-control study was done in the Canadian province of Saskatchewan between 1984 and 1985 among residents who were 65 years of age or older and who were not receiving other drugs thought to affect bone mass. There were 905 hip fractures identified from hospital discharge records and 5137 population controls matched for age, sex, and calendar year. Drug use was ascertained from computerised pharmacy records. Risk of hip fracture decreased significantly with increasing duration of current thiazide use: relative risk (95% confidence interval) of 1.2 (0.9-1.5) for less than 2 years use, 0.8 (0.7 1.0) for use of 2-5 years, and 0.5 (0.3-0.7) for 6 or more years. In contrast, there was no such trend for use of other antihypertensive-diuretic drugs (relative risk 0.9 [0.6-1.3] for use of 6 or more years). This protective effect was not altered by age, sex, nursing home residence, previous hospital admission, or use of other antihypertensive-diuretic drugs or psychotropic drugs. Medical record review for a sample of 235 cases suggested this finding was not due to confounding by body mass, ambulatory status, functional status, or dementia. These results support the hypothesis that thiazides protect against osteoporosis in elderly people. PMID- 2564507 TI - Amoxycillin plus tinidazole for Campylobacter pylori gastritis in children: assessment by serum IgG antibody, pepsinogen I, and gastrin levels. AB - 32 children (mean age 12 years, range 6-18) with non-specific abdominal pain and Campylobacter pylori positive gastritis received a six week course of daily oral amoxycillin (50 mg/kg) and tinidazole (20 mg/kg). Before treatment and one month after stopping treatment, endoscopic biopsy samples were taken from the antral mucosa and serum C pylori IgG antibody, pepsinogen I, and gastrin levels were measured in fasting blood samples. One month after treatment 30 children (94%) were cleared of C pylori and gastritis had resolved in 27 (84%) and was improved in the remaining 5. Serum IgG, pepsinogen I, and gastrin levels were significantly decreased after treatment. Of 12 children assessed at six months, 9 remained free of C pylori. Increases or decreases in IgG level indicated clearance or recurrence, respectively, of C pylori. PMID- 2564508 TI - Inhibition of platelet aggregation by abnormal high density lipoprotein particles in plasma from patients with hepatic cirrhosis. AB - ADP-induced aggregation of normal washed platelets was measured by nephelometry in the presence of plasma high density lipoprotein (HDL) from normal subjects and from 30 patients with hepatic cirrhosis. HDL, at one-eighth of its plasma concentration, inhibited platelet aggregation; the effect of cirrhotic HDL (40% [SD 29%] inhibition) was significantly greater than that of normal HDL (16% [11%]). The mean apolipoprotein E content of cirrhotic HDL was significantly higher than that of normal HDL, and strongly inhibitory HDL contained twice as many apolipoprotein-E-rich particles as weakly inhibitory HDL. Inhibition of platelet aggregation was correlated with the apolipoprotein E content of HDL from patients with cirrhosis. PMID- 2564509 TI - Polymerase chain reaction for detection of dissemination in gastric lymphoma. AB - The polymerase chain reaction (PCR) was used to detect malignant lymphoma cells with the bcl-2 gene rearrangement in the peritoneal washings and bone marrow of a patient with an apparently localised gastric lymphoma. After four courses of cytotoxic drug treatment the cells could no longer be detected in either site. PCR is a useful addition to the staging investigations of non-Hodgkin lymphoma and can also be used to monitor response to treatment. PMID- 2564510 TI - Stem cells in neoplasia. PMID- 2564511 TI - Undergraduate general practice. PMID- 2564512 TI - Polyarticular gout. PMID- 2564513 TI - Acetabular development after closed reduction of congenital dislocation of the hip. PMID- 2564514 TI - Peptide regulatory factors: a new Lancet series. PMID- 2564515 TI - Peptide regulatory factors: multifunctional mediators of cellular growth and differentiation. PMID- 2564516 TI - Atrial natriuretic peptide. An important factor in sodium and blood pressure regulation. PMID- 2564517 TI - Rising mortality from motoneuron disease in the USA, 1962-84. AB - From 1962 to 1984, age-specific mortality for motoneuron disease (MND) in the United States rose in all demographic groups over the age of 40. The increase was seen in both men and women, and both whites and non-whites, and was most pronounced in the elderly (eg, 378% in white women aged 80-84 years). Men were at 50% higher risk than women, and whites had twice the risk of non-whites. These increases may reflect an improvement in case ascertainment, but they also seem to show a true rise in the incidence of MND, particularly among the elderly. Such an increase suggests an environmental aetiology. PMID- 2564518 TI - A sociologist looks at the GMC. PMID- 2564519 TI - Called to account. Surgery. Orthopedics. PMID- 2564520 TI - Population intervention. PMID- 2564521 TI - Secondary prevention of coronary heart disease. PMID- 2564522 TI - Halothane anaesthetic and the ozone layer. PMID- 2564523 TI - Maternal hepatitis B virus DNA in mother-infant transmission. PMID- 2564524 TI - Early detection of breast cancer in developing countries. PMID- 2564526 TI - Management of spontaneous rupture of the membranes and no uterine activity in healthy primigravidae after 34 weeks' gestation. PMID- 2564525 TI - Warfarin to prevent thromboembolism in chronic atrial fibrillation. PMID- 2564527 TI - Treatment of refractory ulcerative colitis with cyclosporin enemas. PMID- 2564528 TI - Desferrioxamine content of aluminum. PMID- 2564529 TI - Simple metabolic bed for paediatric use. PMID- 2564530 TI - Effects of temperature on reactivity of plasma in recombinant HIV-1 competitive assay. PMID- 2564531 TI - Racial differences in gallbladder motor function. PMID- 2564532 TI - Anorectal varices, haemorrhoids, and portal hypertension. PMID- 2564533 TI - Lovastatin and elevated creatine kinase: results of rechallenge. PMID- 2564534 TI - Bone marrow immunoscintigraphy. PMID- 2564535 TI - Activity of thiocarbamazine in patients dually infected with Schistosoma and Onchocerca volvulus. PMID- 2564536 TI - Premarital sex in rural India. PMID- 2564537 TI - Rapid diagnosis of malaria. PMID- 2564538 TI - Oral metolazone plus frusemide for home therapy in patients with refractory heart failure. PMID- 2564539 TI - Adjuvant systemic treatment for breast cancer. PMID- 2564540 TI - Specific IgE-antibodies to Bordetella pertussis antigens during whooping cough. PMID- 2564541 TI - Long-term hepatotoxicity of tacrine. PMID- 2564542 TI - Morbidity of very-low-birthweight infants. PMID- 2564543 TI - CD4 is not the membrane receptor for HHV-6. PMID- 2564544 TI - Parvovirus-associated thrombocytopenic purpura. PMID- 2564545 TI - Dopaminergic mechanism for caffeine-induced decrease in fertility? PMID- 2564546 TI - Neurological decompression sickness. PMID- 2564547 TI - Ocular fundus lesions in divers. PMID- 2564548 TI - Paradoxical systemic embolisation through a patent foramen ovale. PMID- 2564549 TI - C-reactive protein and serum agglutination in vivo of intravenous fat emulsions. PMID- 2564550 TI - Tamoxifen and endometrial cancer. PMID- 2564551 TI - Sniffer dogs in the melanoma clinic? PMID- 2564552 TI - Warning against use of intrathecal mitoxantrone. PMID- 2564553 TI - Immunosuppressive properties of cyclosporin metabolites. PMID- 2564554 TI - Detumescence by exercise bicycle. PMID- 2564555 TI - Increase in resistance to penicillin in pneumococci in Spain. PMID- 2564556 TI - Transfusion associated graft-versus-host disease in autologous bone marrow transplantation. PMID- 2564557 TI - Control of life-threatening bleeding from caput medusae by umbilical vein embolisation. PMID- 2564558 TI - Crohn's disease in a married couple and their four children. PMID- 2564559 TI - Impregnated via a bullet? PMID- 2564561 TI - Have an HIV test today. PMID- 2564560 TI - Delayed birth injuries claim. PMID- 2564562 TI - Mitral valve prolapse as a risk factor for infective endocarditis. AB - The frequency of mitral valve prolapse was assessed in 48 patients with mitral valve endocarditis and in 96 controls matched for age and sex, attending a routine family screening clinic or having surgery of the limbs. The frequency of mitral valve prolapse in cases with endocarditis (9 of 48 patients) was more than three times that in controls (6 of 96) (odds ratio 3.5; 95% confidence interval [CI] 1.1-10.5). When patients with rheumatic heart disease, an established risk factor for infective endocarditis, were excluded from the study group, patients were nearly six times more likely to have infective endocarditis than were controls (odds radio 5.7; 95% CI 1.8-18.4). However, a higher risk of infective endocarditis was seen only in the subjects with mitral valve prolapse and a previously known systolic murmur (odds ratio 14.5; 95% CI 1.7-125). The results indicate that mitral valve prolapse constitutes a true risk factor for infective endocarditis only when associated with the presence of a precordial systolic murmur. PMID- 2564564 TI - Naloxone applied to conjunctiva as a test for physical opiate dependence. AB - Significant pupillary dilatation in response to conjunctivally applied naloxone was demonstrated with polaroid photography in 46 opiate addicts on maintenance methadone prescription. No pupillary response was seen to an identical procedure in 17 non-addict subjects who had been given an opiate premedication before elective surgery. This pupillary response to naloxone could be used as a test to identify the physically dependent opiate user. PMID- 2564563 TI - Comparison of fluconazole and ketoconazole for oropharyngeal candidiasis in AIDS. AB - In a randomised, double-blind study the efficacy and toxicity of oral fluconazole 50 mg daily and ketoconazole 200 mg daily were compared for the treatment of oropharyngeal candidiasis in patients with acquired immunodeficiency syndrome (AIDS) and AIDS-related complex (ARC). 20 episodes (18 patients) were treated with fluconazole and 20 episodes (19 patients) with ketoconazole. Pretreatment clinical features and laboratory test results were similar in both groups. 17 episodes (85%) in the fluconazole group and 16 (80%) in the ketoconazole group could be evaluated. There was clinical cure at the end of therapy in all fluconazole-treated and 12 of 16 (75%) ketoconazole-treated episodes. Cultures were negative at the end of therapy in 87% of the fluconazole group and 69% of the ketoconazole group. 1 patients stopped taking fluconazole because of severe nausea. 1 of 18 fluconazole-treated and 4 of 19 ketoconazole-treated patients had transient rises in alanine or aspartate aminotransferase. Fluconazole seemed more effective than ketoconazole in the treatment of oral thrush among AIDS and ARC patients. PMID- 2564565 TI - Diagnostic value of ascites adenosine deaminase in tuberculous peritonitis. AB - The value of ascitic fluid adenosine deaminase activity in distinguishing tuberculosis from other causes of ascites was examined in a retrospective study of 41 patients with bacteriologically confirmed tuberculous peritonitis and 41 control patients, matched for age and sex, with ascites of other causes (12 alcoholic cirrhosis, 5 cryptogenic cirrhosis, 12 malignant disorders, 3 pancreatitis, and 9 miscellaneous causes). The mean ascites adenosine deaminase activity was 99.8 (SD 49.1) in tuberculous patients and 14.8 (8.4) U/l in control patients (p less than 0.0001). A cutoff of 32.3 U/l had a sensitivity of 95% and specificity of 98% in distinguishing between the two groups. In a subsequent prospective study of 64 patients with ascites, 11 were found to have tuberculosis. Of the others, 23 had cirrhosis (18 alcoholic, 5 cryptogenic), 17 malignant disorders, 3 pancreatitis, 5 cor pulmonale, 3 congestive cardiac failure, 1 systemic mastocytosis, and 1 renal failure and hypothyroidism. The mean ascites adenosine deaminase activity was 112.6 (45.0) U/l in the patients with tuberculous ascites and 16.3 (36.7) U/l (p less than 0.0001) in those with ascites of other causes. In this study, adenosine deaminase had a sensitivity of 100% and specificity of 96% in discriminating tuberculosis from other causes of ascites. These findings suggest that the ascitic fluid adenosine deaminase activity may be used to identify patients in whom the diagnosis of abdominal tuberculosis must be pursued. PMID- 2564566 TI - Induction of graft-versus-host disease after autologous bone marrow transplantation. AB - To induce graft-versus-host disease (GvHD) in patients undergoing autologous bone marrow transplantation (BMT), five consecutive patients with non-Hodgkin lymphoma or Hodgkin's disease in resistant relapse were treated with cyclophosphamide and total body irradiation or busulphan and cyclophosphamide, followed by autologous BMT. The patients received cyclosporin 1 mg/kg per day intravenously for 28 days after BMT. Histologically proven grade II acute GvHD of the skin developed in all the patients at a median of 11 days (range 9-13) after BMT. One patient died and the GvHD resolved in 1-3 weeks in the four remaining patients--spontaneously in two and with corticosteroid treatment in two. Lymphocytes from one patient obtained at the time of GvHD were cytotoxic for the patient's own pretransplant lymphocytes. Cytotoxicity was blocked by antibodies directed against class II histocompatibility (HLA-DR or Ia) antigens. Cyclosporin-induced GvHD after autologous BMT resembles mild GvHD after allogeneic grafting. This syndrome appears to be mediated by autoreactive Ia-specific lymphocytes. PMID- 2564567 TI - Trauma and dystonia. PMID- 2564568 TI - Behcet's disease. PMID- 2564569 TI - Transferring diabetic patients to human insulin. PMID- 2564570 TI - Exercise training, fitness, and asthma. PMID- 2564571 TI - Diversion colitis. PMID- 2564572 TI - Post-receptor signalling pathways. PMID- 2564573 TI - The origins of intravenous fluid therapy. PMID- 2564574 TI - Congenital malformations and oral poliovirus vaccination during pregnancy. AB - In February, 1985, mass vaccination with live oral poliovirus vaccine (OPV) was started during a poliomyelitis outbreak in Finland. Pregnant women were advised to take the vaccination. Judged as the rate of reported malformations, OPV during early pregnancy had no harmful effects on fetal development. PMID- 2564575 TI - Resource allocation and bioethics. PMID- 2564576 TI - Acute haemorrhagic gastritis controlled by omeprazole. PMID- 2564577 TI - Gastric carcinoid tumours in patients with Zollinger-Ellison syndrome on long term omeprazole. PMID- 2564578 TI - Buspirone in treatment of premenstrual syndrome. PMID- 2564579 TI - Comparative virulence of respiratory syncytial virus subgroups A and B. PMID- 2564580 TI - Diagnosis of herpes simplex encephalitis by single photon emission tomography. PMID- 2564581 TI - Detection of 14;18 chromosomal translocation in paraffin-embedded lymphoma tissue. PMID- 2564582 TI - Globus. PMID- 2564583 TI - Aluminium and the nucleus of nerve cells. PMID- 2564584 TI - Aluminium, hot water tanks, and neurobiology. PMID- 2564585 TI - Aluminium from a coffee pot. PMID- 2564586 TI - International prostitutes and transmission of HIV. PMID- 2564587 TI - Oral contraceptives and breast cancer. PMID- 2564588 TI - Oral contraception and genital tract malignancy. PMID- 2564589 TI - Cervical cancer deaths in young women. PMID- 2564590 TI - Endocervical brush device. PMID- 2564591 TI - Cardiac hypertrophy in old age. PMID- 2564592 TI - Histopathologists, malignancies, and undefined high-power fields. PMID- 2564593 TI - Human monoclonal antibody to ganglioside GM2 for melanoma treatment. PMID- 2564594 TI - Insulin pen injectors. PMID- 2564595 TI - Trials of dipyrone in Thailand. PMID- 2564596 TI - Israel: doctors and defence forces. PMID- 2564597 TI - Academic obstetrics and gynaecology. PMID- 2564598 TI - Occupations, deaths, and coroners' inquests. PMID- 2564599 TI - Borrelia burgdorferi infection in UK workers at risk of tick bites. PMID- 2564600 TI - Long-term follow-up of immunomodulation in treatment of HTLV-I-associated myelopathy. PMID- 2564601 TI - Dietary fat guidelines for men and women. PMID- 2564602 TI - Sarcosporidiosis revealed in sputum. PMID- 2564603 TI - Plasma D-dimer and pulmonary embolism. PMID- 2564604 TI - Germ cell injury and childhood leukaemia clusters. PMID- 2564605 TI - Childhood leukaemia mortality before 1970 among populations near two US nuclear installations. PMID- 2564606 TI - Intestinal-type gastric carcinoma and colonic carcinoma: a common pathogenesis? PMID- 2564607 TI - Fatal listeria meningitis in immunosuppressed patient. PMID- 2564608 TI - Listeriosis in immunocompromised pregnancy. PMID- 2564609 TI - Medical defence and crown indemnity. PMID- 2564610 TI - AIDS education. PMID- 2564611 TI - Functional characterization of beta-adrenoceptor subtypes in rabbit right atria. AB - The advent of radioligand binding studies has allowed the classification of receptor subtypes in various tissues. However, the presence of a receptor subtype in a heterogenous tissue does not insure that the receptor has a significant physiological role. beta 1- and beta 2-Adrenoceptors have been reported to coexist in the rabbit right atria. The purpose of the present investigation was to determine the physiological role of beta-adrenoceptor subtypes in catecholamine-induced chronotropic responses in the rabbit right atria through comparison of data from functional and radioligand binding studies. Rank order of potency was determined using isoproterenol, epinephrine and norepinephrine for both chronotropic and inotropic responses in the rabbit right atria and right ventricular papillary muscles, respectively. These studies indicated that the beta 1-adrenoceptor was primarily responsible for catecholamine-induced responses. Next, the beta 1-selective antagonist, atenolol, was found to inhibit the chronotropic responses of the nonselective beta-agonist, isoproterenol, and the beta 2-selective agonist, terbutaline, to the same extent. These data indicate that terbutaline produces its chronotropic effects in the rabbit right atria through stimulation of beta 1-, not beta 2-adrenoceptors. Finally, competition studies for [125I]iodocyanopindolol and the relatively selective beta 1- and beta 2-adrenoceptor antagonists (ICI 89406 and ICI 118551, respectively) indicated that the ratio of beta 1- to beta 2-adrenoceptor subtypes is 6:1. It is concluded that while both receptors may be present in the rabbit right atria, the beta 1-adrenoceptor is the predominant subtype both in density and physiological significance, while the beta 2-adrenoceptor plays little, if any role, in the chronotropic responses induced by catecholamines. PMID- 2564612 TI - Validation of transport measurements in skeletal muscle with N-13 amino acids using a rabbit isolated hindlimb model. AB - We are studying the transport of C-11 and N-13 labeled amino acids in tumor bearing rabbits to determine the role of amino acid transport in the pathogenesis of muscle wasting in cancer. To validate a new, in vivo, method for measuring transport in skeletal muscle with these compounds, an isolated hindlimb model was developed in rabbits. The limb was perfused with a non-recirculating, normothermic, constant pressure system and a cell-free perfusate. Hemodynamic and metabolic parameters were measured during the first 75 min. of perfusion and found to remain normal and stable. Flow varied directly with perfusion pressure over the normal range of resting flows in the intact rabbit hindlimb. Time activity curves (TAC's) were recorded from the medial thigh following bolus co injection of L-[amide N-13] glutamine or N-13 L-glutamate with Tc-99m human serum albumin (HSA) into the femoral artery. Regional plasma flow was determined from the Tc-99m data. The N-13 TAC's consistently manifested a three-phased washout with half times of approximately 30 sec., 5 min. and 2 hr. Capillary and cellular transport parameters were computed from the N-13 data using a double barrier, single capillary model of capillary and cellular transport and assuming that the three washout components result, respectively, from tracer throughput, extraction into the interstitial space and extraction into the intracellular space. This interpretation was validated and the sensitivity of the technique to transport processes demonstrated by examining the effects on the N-13 TAC's and computed transport parameters of several factors known to influence cellular transport of amino acids, viz., the insulin concentration, amino acid concentration and pH of the perfusate. Time-activity curves and transport parameters for N-13 L-glutamine in the isolated limb were very similar to those observed in the intact rabbit hindlimb, suggesting that studies in the perfused model are indicative of amino acid transport in vivo. The methodology described here is especially well suited for studying the specific effects on transport of factors which influence amino acid metabolism in skeletal muscle (e.g., hormones and monokines). PMID- 2564613 TI - Effects of specific alpha-adrenoceptive agents on extraneuronal uptake (uptake2) of isoproterenol in perfused rat heart. AB - Effects of specific alpha-adrenoceptive agents (alpha 1-agonist, alpha 1 antagonist, alpha 2-agonist and alpha 2-antagonist) on the extraneuronal accumulation of 3H-isoproterenol in the perfused rat heart were examined. The extraneuronal accumulation of 3H-isoproterenol in the hearts perfused with 3H isoproterenol (10(-6)M) under COMT inhibition by tropolone (10(-4)M) was about 6 times higher than that of intact COMT. The increase in the accumulation by COMT inhibition was regarded as 100% and the effects of specific alpha-adrenoceptive agents on the accumulation was evaluated. alpha 1-agonists, methoxamine and phenylephrine, did not affect the accumulation. alpha 1-antagonists, prazosin, bunazosin and YM-12617, significantly decreased the accumulation of 3H isoproterenol and these IC50 values were 2 x 10(-6)M, 3.5 x 10(-6)M and 2.3 x 10( 5)M, respectively. alpha 2-agonists, clonidine and guanabenz, significantly reduced the accumulation and these IC50 values were 3.4 x 10(-5)M and 2.9 x 10( 7)M, respectively. The alpha 2-antagonist, yohimbine, did not affect the accumulation. The present experiments clearly demonstrated that the tested alpha 1-antagonists and alpha 2-agonists inhibited uptake2 in rat heart but the tested alpha 1-agonists and an alpha 2-antagonist did not inhibit it. PMID- 2564614 TI - Inhibition of hepatic cholesterol synthesis by the alpha 1-adrenoceptor blocker doxazosin in the hypercholesterolemic golden hamster. AB - The effect of treatment with the alpha 1-specific adrenoceptor blocker, Doxazosin, on lipid parameters was studied in male Golden hamsters fed a cholesterol-enriched diet. Within 1 week the Doxazosin-treated animals had a lower plasma (-12%) and hepatic (-30%) cholesterol content than the cholesterol fed controls. De novo cholesterol synthesis in the liver was lowered by 39% in the Doxazosin-treated animals. These data indicate that the reported beneficial effect of alpha 1-blockade on plasma cholesterol levels may be due to lowering of the hepatic cholesterol synthesis. PMID- 2564616 TI - Chronic electroconvulsive shock and neurotransmitter receptors--an update. AB - Electroconvulsive shock (ECS) produces many neurochemical alterations which may be related to its efficacy in the treatment of different psychiatric disorders. This review focuses particularly on experimental findings of CNS receptor changes in animals following chronic ECS and relates them to neurotransmitter and behavioral changes. Also, the pharmacological effect of other antidepressant treatment are compared. Possible mechanisms of action are discussed. PMID- 2564615 TI - Behavioral recovery after irreversible inactivation of D-1 and D-2 dopamine receptors. AB - Irreversible inactivation of both D-1 and D-2 dopamine (DA) receptors by N ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) resulted in complete loss of stereotypy response to R-(-)-N-propylnorapomorphine (NPA; 0.1-1.0 mg/kg, s.c.) 24 hr later. Stereotyped sniffing recovered much more rapidly than oral behaviors. The D-2 antagonist sulpiride (200 mg/kg) and the putatively nonselective antagonist cis-flupenthixol (2 mg/kg), administered prior to EEDQ, prevented the loss of NPA-induced sniffing but only partially protected against loss of oral behaviors 24 hr later. Complete protection of both behaviors was seen after pretreatment with a combination of sulpiride and the selective D-1 antagonist SCH 23390 (1 mg/kg); pretreatment with the selective D-1 antagonist SCH 23390 alone, however, did not modify the rate of recovery of either behavioral response. The results suggest that either different populations of DA receptors mediate expression of these behaviors or stimulation of a small fraction of the total DA receptor pool may be sufficient to elicit sniffing but not oral responses. Furthermore, maintaining a normal complement of D-2 rather than D-1 receptors appears to be a critical determinant for the elicitation of these behaviors. PMID- 2564617 TI - [3H]L-657,743 (MK-912): a new, high affinity, selective radioligand for brain alpha 2-adrenoceptors. AB - L-657,743 (MK-912), a highly potent and selective alpha 2-adrenoceptor antagonist was tritiated to a high specific activity and its binding characteristics to brain tissue were determined. The specific binding of [3H]L-657,743 to rat cerebrocortex was saturable, reversible, and dependent on tissue concentration. In saturation studies, [3H]L-657,743 binding was resolved into two high affinity components exhibiting Kd values of 86 pM and 830 pM with densities of 82 fmol/mg protein and 660 fmol/mg protein, respectively. Based on the binding potencies of a variety of compounds with differing receptor selectivities, the sites labeled by [3H]L-657,743 were characteristic of alpha 2-adrenoceptors. In contrast to alpha 2-antagonists, alpha 2-agonists displayed shallow competition curves. In the presence of 100 microM GTP, Gpp(NH)p or 150 mM NaCl, the competition curve for epinephrine was shifted to the right, whereas that for yohimbine was unaffected. In studies utilizing human cerebrocortical tissue, [3H]L-657,743 also bound with high affinity to sites characteristic of alpha 2-adrenoceptors. PMID- 2564618 TI - Hypothalamic growth hormone-releasing factor (GRF) participates in the negative feedback regulation of growth hormone secretion. AB - Effects of growth hormone (GH) excess on immunoreactive hypothalamic GH-releasing factor (GRF) and somatostatin (SRIF) were studied in rats. Hypothalamic GRF content significantly reduced after 7-day daily treatment with 160 micrograms of rat GH or after inoculation of GH-secreting rat pituitary tumors, MtT-F4 for 9 or 13 days and GH3 for 3 months. Basal and 59 mM K+-evoked release of GRF from incubated hypothalami diminished, more than the content, by 43-51% in MtT-F4 tumor- or by 67-83% in GH3 tumor-bearing rats. In contrast, there was a small but significant increase in content or release of SRIF in rats harboring the GH3 or MtT-F4 tumor, respectively. These results indicate the existence of a negative feedback loop via hypothalamic GRF as well as SRIF in control of GH secretion. PMID- 2564619 TI - High affinity binding sites for basic fibroblast growth factor in rat hepatic plasma membranes. AB - The binding of [125I]-recombinant basic FGF (rec bFGF) to rat hepatic plasma membranes was investigated. [125I] rec bFGF bound to an apparent single class of high affinity binding sites (KD = 69 pM; Bmax = 9.61 fmoles/mg proteins). The absence of low affinity sites was confirmed by the inability of sulphated polysaccharides and heparinase to interfere with FGF binding. A good correlation existed between the ability of bovine pituitary-derived bFGF, rec bFGF and bovine brain-derived aFGF to displace [125I]rec bFGF from these binding sites and their in vitro potency on bovine aortic endothelial cell proliferation. PMID- 2564620 TI - Chronic, but not acute, administration of morphine alters antiopiate (Tyr-MIF-1) binding sites in rat brain. AB - Opiate addiction could involve a change in the binding of endogenous antiopiates. A candidate for such a role is Tyr-MIF-1 (Tyr-Pro-Leu-Gly-NH2), a brain peptide that can antagonize exogenous and endogenous opiates and bind to opiate receptors. Its primary action, however, may be through its own binding site in brain, which we now report is altered by chronic administration of morphine. Rats given morphine pellets had reduced binding of both iodinated and tritiated Tyr MIF-1 on day 5, when substantial tolerance is evident. In contrast, mu and delta opiate receptors were increased. Acute injection of an analgesic dose of morphine did not reduce Tyr-MIF-1 binding, indicating that chronic administration is required for the change. These findings open new approaches to the study of addiction by focusing on antiopiate activity. PMID- 2564621 TI - The current status of opioid research on gastrointestinal motility. AB - Apparently conflicting data on opioid effects on gastrointestinal motility have been reported in the literature. The current status is reviewed and an attempt is made to find a common denominator to discrepant results by suggesting functionally contrasting opioid systems modulating the same physiological functions. Upon superimposition, these contrasting systems might result in opposite opioid effects dependent on the actual functional balance between the systems at the time of drug administration. Inhibitory neuromodulation at multiple sites leading to either inhibition or disinhibition by opioids may serve as a common basis of their contrasting effects. This interpretation, though consistent with most of the currently available data, is still a working hypothesis. PMID- 2564622 TI - Transcriptional control of adrenal catecholamine and opiate peptide transmitter genes. AB - In the rat, decreasing transsynaptic activity through adrenal denervation, nicotinic receptor blockade, or explanation is associated with an increase in preproenkephalin mRNA, enkephalin prohormone and peptide. In contrast, catecholamine pathways remain unchanged under similar conditions. Since it is not known whether changes in messenger RNA result from stabilization or increased synthesis, we exploited transcription 'run-on' assays to measure the rate of transmitter gene read out. Tyrosine hydroxylase message (TH-mRNA) was found to be the most abundantly produced transcript in the unmanipulated control rat adrenal medulla. TH-mRNA was produced in excess of twice the rate of transcription of the structural gene beta-actin. In contrast, preproenkephalin transcription occurred at a much lower rate (60% of the actin gene and only 25% of tyrosine hydroxylase gene transcription). All transcripts were inhibited by the polymerase II inhibitor, alpha-amanitin. After two days in explant culture, the rate of enkephalin transcription increased approximately 2-fold (to the same level as actin transcription); while tyrosine hydroxylase transcriptional activity fell to 30% of actin level. To analyze cellular mechanisms, explants were depolarized with potassium chloride. Enkephalin gene transcription was observed to be 2.5 fold less when grown under depolarizing conditions (50 mM KCl) than in control explants. On the other hand, tyrosine hydroxylase gene read-out was unchanged, similar to results obtained when TH catalytic activity was measured. These data indicate that membrane depolarization can selectively regulate expression of a transmitter gene product and are consistent with a proposed transsynaptic regulatory mechanism controlling biosynthesis of adrenal opiate peptides.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2564623 TI - Differences in the effects of membrane depolarization on levels of preprosomatostatin mRNA and tyrosine hydroxylase mRNA in rat sympathetic neurons in vivo and in culture. AB - Regulation of preprosomatostatin mRNA and tyrosine hydroxylase mRNA were examined in sympathetic neurons of the rat superior cervical ganglion (SCG). Surgical denervation of the adult SCG increased ganglion levels of preprosomatostatin (SS) mRNA more than 11-fold, and levels of the mRNA remained elevated 14 days after surgery. By contrast, denervation decreased levels of tyrosine hydroxylase (TH) mRNA. Potassium- or veratridine-induced membrane depolarization of cultured neonatal sympathetic neurons decreased levels of SS mRNA but elevated levels of TH mRNA. Sodium channel blockade with tetrodotoxin prevented the effects of veratridine on SS and TH mRNAs. In toto these observations suggest that transsynaptic nerve impulse activity and sympathetic neuron membrane depolarization decrease SS synthesis but increase TH synthesis at the mRNA level. Thus nerve impulse activity may alter the relative levels of different transmitters co-expressed in the same neuronal population by inhibiting levels of some species of mRNA while simultaneously stimulating levels of others. PMID- 2564624 TI - [The oldest school on the Don]. PMID- 2564625 TI - AIDS is changing the world. Report on the Fourth International Conference on AIDS, Stockholm, Sweden 12-16 June, 1988. PMID- 2564626 TI - Opioids a better treatment for acute than tardive akathisia: possible role for the endogenous opiate system in neuroleptic-induced akathisia. AB - Patients who are on neuroleptics with acute akathisia improve more with opioid therapy than patients who are off neuroleptics with tardive akathisia. Since tardive akathisia usually occurs after long term neuroleptic exposure, we propose that this difference in therapeutic response is due to permanent changes in receptor sensitivity that arise from such prolonged exposure. We have previously described a patient with severe acute akathisia whose motor restlessness was totally suppressed by opioid therapy. This improvement was rapidly reversed by the opiate receptor blocker naloxone. This suggests that the endogenous opiate system is involved in the pathogenesis of neuroleptic-induced akathisia. PMID- 2564627 TI - Maserlike nonlinear scatter from human breath, a surface-enhanced far infrared scatter effect. AB - A cotton tip applicator was utilized to stimulate far IR scatter (emission) from human breath. The reticulated cotton tip was vibrated at 200 Hz, the acoustic frequency that matches the average vibration frequency of common species of mosquitoes. Mosquitoes have open resonator dielectric sensillae (spines) on their antenna that match the 643 cm-1 (15.5) Cabannes line in the 667 cm-1 region of CO2 rotation line, acoustic Stokes Brillouin and Stokes Raman lines are noted. Implications of such maserlike (coherent or partial coherent) scatter lines to insect control and disease diagnostics are discussed. PMID- 2564628 TI - Role of differential drug uptake, efflux, and binding of etoposide in sensitive and resistant human tumor cell lines: implications for the mechanisms of drug resistance. AB - In order to study the mechanism of etoposide (VP-16) resistance in human tumor cells and to assess the role of P-170 glycoprotein in VP-16 accumulation, we have examined the uptake and efflux of VP-16 in both sensitive and multidrug-resistant MCF-7 human breast and HL60 human promyelocytic leukemia cells. The drug resistant cells, MCF-7/ADR and HL60/ADR, were selected for resistance to adriamycin and were 200- to 250-fold resistant to VP-16. Whereas MCF-7/ADR cells overexpress the P-170 glycoprotein and show the multidrug-resistant phenotype, HL60/ADR cells do not overexpress the P-170 glycoprotein. Although there was a 2 fold decrease in accumulation of VP-16 in MCF-7/ADR cells, this decrease did not correlate with a 250-fold resistance to the drug. VP-16 efflux was rapid and almost complete from MCF-7 cell lines and it was decreased at 4 degrees. Further, there was a significant increase in VP-16 accumulation in the MCF-7/ADR cells in the presence of glucose-free medium supplemented with sodium azide. However, no change in the pattern of VP-16 efflux was observed. Under these conditions, addition of glucose caused release of VP-16 from MCF-7/ADR cells, suggesting energy-dependent modifications in the drug binding. Coincubation of vincristine with VP-16 also increased the drug accumulation and decreased the rate of efflux of VP-16 in both sensitive and resistant MCF-7 cells, suggesting that vincristine and VP-16 may compete for similar binding and efflux mechanisms in these cell lines. In contrast, daunorubicin increased VP-16 accumulation only in the sensitive MCF-7 cell line, whereas the efflux rate of VP-16 was not significantly changed in either cell line. HL60 sensitive cells accumulated 4- to 5-fold more VP-16 than the resistant subline. Both sensitive and resistant cells showed an important noneffluxable pool of the drug, 3-fold larger for sensitive cells (79 +/- 12 versus 25 +/- 2 pmol of VP-16/mg of protein, for sensitive and resistant cells, respectively). The efflux of VP-16 was temperature dependent only in sensitive cells. VP-16 accumulation in HL60/ADR cells was increased in glucose free medium supplemented with sodium azide; however, the noneffluxable pool of VP 16 was not significantly changed. In contrast, although these conditions had no effect on the drug accumulation in the parental line, they caused a decrease in the noneffluxable pool of VP-16, suggesting an energy-dependent binding and retention of VP-16.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2564629 TI - Beta 1- and beta 2-adrenergic receptor-mediated adenylate cyclase stimulation in nonfailing and failing human ventricular myocardium. AB - Prenalterol (beta 1-agonist), denopamine (beta 1-agonist), and zinterol (beta 2 agonist) were partial agonists of adenylate cyclase (AC) stimulation in human ventricular myocardium obtained from nonfailing chambers whose beta 1/beta 2 receptor subtype ratio was approximately 80/20. At a concentration less than its low affinity (beta 2) Kl, betaxolol, a highly selective beta 1-antagonist, inhibited isoproterenol (non-selective agonist), denopamine, and prenalterol stimulation of AC, indicating that isoproterenol, denopamine, and prenalterol are all capable of stimulating AC through beta 1-receptor activation. At a concentration less than its low affinity (beta 1) Kl, ICI 118,551, a highly selective beta 2-agonist, inhibited both isoproterenol and zinterol stimulation of AC, indicating that isoproterenol and zinterol stimulate AC through beta 2 receptors. Zinterol stimulation of AC was mediated entirely by beta 2-receptors, inasmuch as 10(-7) M betaxolol had no effect on the zinterol dose-response curve and ICI 118,551 produced a degree of blockade (KB = 5.2 +/- 1.6 X 10(-9) M), consistent with the beta 2-receptor Kl of the latter (2.0 +/- .4 X 10(-9) M, p, not significant). In nonfailing myocardium, analysis of beta 1 versus beta 2 stimulation by the nonselective agonist isoproterenol revealed that the numerically small (19% of the total) beta 2 fraction accounted for the majority of the total adenylate cyclase stimulation. In failing ventricular chambers with a beta 1/beta 2 receptor subtype ratio reduced from 82/19 (nonfailing) to 64/36 (p less than 0.001) and a beta 1-receptor density reduced by 61% (p less than 0.001), maximal denopamine stimulation was reduced by 49% (p less than 0.001). Moreover, in preparations from failing heart, the component of denopamine stimulation that was inhibited by 10(-7) M betaxolol (beta 1 component) was reduced by 77% (p less than 0.05). Finally, in preparations derived from failing ventricular myocardium, beta 2-receptor density was not significantly decreased, but zinterol stimulation of AC was reduced by 32% (p less than 0.05). We conclude that heart failure results in subsensitivity to both selective beta 1 and beta 2 stimulation of adenylate cyclase, with beta 1 subsensitivity due to selective beta 1 receptor down-regulation and beta 2 subsensitivity due to partial uncoupling of beta 2 receptors from subsequent events in the beta 2-adrenergic pathway. PMID- 2564630 TI - Beta-adrenergic desensitization reduces the sensitivity of adenylate cyclase for magnesium in permeabilized lymphocytes. AB - Magnesium modulates hormone-sensitive adenylate cyclase activation. In the present studies, we have examined the magnesium requirement of beta-adrenergic stimulated adenylate cyclase activity in permeabilized human lymphocytes. Following isoproterenol pretreatment, under conditions that lead to homologous beta-adrenergic desensitization, the EC50 of magnesium for beta-adrenergic stimulated adenylate cyclase activity was significantly increased [control; 1.99 (+0.81/-0.57) mM; desensitized; 3.82 (+0.31/-0.29) mM]. Further, when assays were performed at high Mg2+ concentrations following agonist pretreatment, we detected only small and inconsistant reductions in beta-adrenergic-stimulated adenylate cyclase activity and in beta-adrenergic cAMP-dependent protein kinase activity. In contrast, the detection of agonist-induced beta-adrenergic receptor sequestration and alterations in receptor affinity for agonists was not qualitatively affected by changes in magnesium concentrations. The data demonstrate that the functional consequences of beta-adrenergic desensitization may be obscured at high magnesium concentrations. Furthermore, in this system desensitization may be viewed as a beta-receptor-specific reduction in magnesium sensitivity. PMID- 2564631 TI - [3H]idazoxan and some other alpha 2-adrenergic drugs also bind with high affinity to a nonadrenergic site. AB - We compared the pharmacological properties of the alpha 2-adrenergic radioligand [3H]idazoxan with those of [3H]rauwolscine in rat and [3H]yohimbine in human renal cortical membranes. The density of "specific" [3H]idazoxan binding sites (defined by 100 microM tolazoline) was twice as high as that of [3H]rauwolscine in rat kidney and four times as high as that of [3H]yohimbine in human kidney. A variety of structurally different drugs fully competed for specific [3H]rauwolscine and [3H]yohimbine binding, with affinities appropriate for the interaction with alpha 2-adrenergic receptors. Specific [3H]idazoxan binding, however, was only partially competed for by the catecholamines epinephrine and norepinephrine in both tissues. Thus, [3H]idazoxan labels both alpha 2-adrenergic receptors and a nonadrenergic site. Clonidine, B-HT 920, moxonidine, phentolamine, prazosin, yohimbine, dopamine, and serotonin also could not compete for this site. However, UK 14,304, guanabenz, indanidine, tolazoline, oxymetazoline, and SK&F 104,078 competed for the additional [3H]idazoxan sites with affinities similar to those at alpha 2-adrenergic receptors. [3H]idazoxan binding substantially in excess of [3H]rauwolscine or [3H]yohimbine binding was also found in human platelets, myometrium, and erythroleukemia (HEL) cells but not in three cell lines lacking alpha 2-receptors (MDCK, BC3H1, and Jurkat cells). Although we have been unsuccessful thus far in defining the precise nature of the additional [3H]idazoxan binding sites, we hypothesize that these sites may be closely affiliated with alpha 2-adrenergic receptors but clearly distinct from the catecholamine binding site of the receptor. The results indicate that care must be taken in the use of [3H]idazoxan or drugs that are recognized at its nonadrenergic site when studying alpha 2-adrenergic effects and receptor subtypes. PMID- 2564632 TI - Interactions of full and partial agonists with HT29 cell alpha 2-adrenoceptor: comparative study of [3H]UK-14,304 and [3H]clonidine binding. AB - The HT29 cell line expresses alpha 2-adrenoceptors that are negatively coupled to the adenylate cyclase system and is, in this respect, a valuable model for in vitro study of alpha 2-adrenergic receptivity in a tissue from human origin. In these cancerous cells, UK-14,304 is a full agonist of the alpha 2-adrenergic mediated inhibition of the vasoactive intestinal peptide-induced cyclic AMP accumulation, whereas clonidine acts only as a partial agonist. In the present report, we used [3H]UK-14,304 as radioligand and compared its binding characteristics with those of [3H]clonidine in order to better understand the difference between full and partial agonism on the basis of agonist/receptor interactions. [3H]UK-14,304 labeled with high affinity (KD = 0.39 +/- 0.05 nM) a single class of sites having the pharmacological specificity of an alpha 2 adrenoceptor. Comparison of [3H]UK-14,304, [3H]clonidine, and [3H]yohimbine Bmax proved that both 3H-agonists labeled the same number of sites (172 +/- 14 versus 179 +/- 21 fmol/mg of protein), whereas the 3H-antagonist recognized more sites (246 +/- 22 fmol/mg of protein). Inhibition of [3H]yohimbine by the two agonists was consistent with the existence of an heterogeneous population of receptors and analysis of the data according a two-site inhibition model showed 1) that the KiL/KiH ratio was higher for UK-14,304 than for clonidine and 2) that the percentages of high affinity state receptor recognized by both agonists were identical (56 +/- 4% with UK-14,304 and 59 +/- 5% with clonidine). Kinetics of [3H]UK-14,304 and [3H]clonidine binding indicated more complex agonist-receptor interactions than equilibrium data did. Association as well as dissociation of both radioligands appeared to be biphasic, suggesting a relative heterogeneity of 3H-agonist binding sites. Kinetic behavior of [3H]UK-14,304 only differed from that of [3H]clonidine by a much slower dissociation rate and by its ability to induce the formation of a tightly bound component, which corresponded to the formation of a very stable full agonist/receptor/Gi complex. Effects of guanosine 5'-(imido)-triphosphate and GTP on [3H]UK-14,304 and [3H]clonidine binding also proved that the agonists were not similarly sensitive to guanine nucleotides. PMID- 2564633 TI - N-methyl-D-aspartate/glycine and quisqualate/kainate receptors expressed in Xenopus oocytes: antagonist pharmacology. AB - Quantitative pharmacological studies were done to determine the properties of excitatory amino acid receptors expressed in Xenopus oocytes injected with rat brain mRNA. Smooth currents with properties indicative of N-methyl-D-aspartate (NMDA) and quisqualate/kainate receptors were observed in mRNA-injected oocytes. Schild analysis of currents evoked by NMDA indicated that the EAA receptor antagonist D-2-amino-5-phosphonovalerate (D-APV) exerted a competitive block of the oocyte NMDA receptor, because the Schild regression was linear with a slope not significantly different from unity (1.03 +/- 0.025) up to 100 microM D-APV. The pA2 estimated for D-APV antagonism of NMDA currents (5.87 +/- 0.043) was nearly identical to that for D-APV as an L-aspartate antagonist (pA2 = 5.86 +/- 0.073, slope = 0.97 +/- 0.036), suggesting that these two agonists are selective for NMDA receptors in oocytes up to concentrations well above 1 mM. 6-Nitro-7 cyano-quinoxaline-2,3-dione (CNQX) reduced the maximum NMDA response significantly (70% reduction by 15 microM CNQX) but had no effect on the NMDA EC50. CNQX exerted a mixed competitive-noncompetitive block of the glycine site on NMDA receptors; 15 microM CNQX increased the glycine EC50 by 5-fold and reduced the maximum glycine response by 35%. In addition, CNQX exerted a potent and competitive antagonism of currents evoked by kainate. The Schild regression was linear up to 30 microM CNQX with a slope of 1.02 +/- 0.014 and a pA2 of 6.53 +/- 0.029. The block of kainate or NMDA currents by 2 microM CNQX was not voltage dependent. D-APV exerted a weak antagonism of kainate-evoked currents, with a pA2 of 3.39 +/- 0.044, but the slope of the Schild regression was slightly less than 1 (0.90 +/- 0.03). These data demonstrate a clear pharmacological distinction between receptors that mediate the kainate- and NMDA-induced currents and quantify the potency of CNQX and D-APV acting at NMDA/glycine and quisqualate/kainate receptors. The implications of these data for the identification of EAA receptors in oocytes and the classification of neuronal EAA receptors are discussed. PMID- 2564634 TI - The general control activator protein GCN4 is essential for a basal level of ARO3 gene expression in Saccharomyces cerevisiae. AB - The ARO3 gene encodes one of two 3-deoxy-D-arabino-heptulosonate-7-phosphate isoenzymes in Saccharomyces cerevisiae catalyzing the first step in the biosynthesis of aromatic amino acids. The ARO3-encoded 3-deoxy-D-arabino heptulosonate-7-phosphate synthase (EC 4.1.2.15) is feedback inhibited by phenylalanine; its isoenzyme, the ARO4 gene product, is inhibited by tyrosine. Both genes ARO3 and ARO4 are strongly regulated under the general control regulatory system. Cells carrying only one intact isogene are phenotypically indistinguishable from a wild-type strain when grown on minimal medium. The complete functional ARO3 promoter comprises 231 base pairs and contains only one TGACTA binding site for the general control activator protein GCN4. Mutating this element to TTACTA inhibits binding of GCN4 and results in a decreased basal level of ARO3 gene product and slow growth of a strain defective in its isogene ARO4. In addition, ARO3 gene expression cannot be elevated under amino acid starvation conditions. An ARO3 aro4 strain with gcn4 genetic background has the same phenotype of low ARO3 gene expression and slow growth. The amount of GCN4 protein present in repressed wild-type cells therefore seems to contribute to a basal level of ARO3 gene expression. The general control activator GCN4 has thus two functions: (i) to maintain a basal level of ARO3 transcription (basal control) in the presence of amino acids and (ii) to derepress the ARO3 gene to a higher transcription rate under amino acid starvation (general control). PMID- 2564635 TI - New B-lymphocyte-specific enhancer-binding protein. AB - We report the discovery of a new B-lymphocyte-specific enhancer-binding protein. A series of gel retardation assays using fragments that scan the -2172 to -1180 region of the major histocompatibility complex class II gene E alpha reveal a site (W) that serves as the recognition sequence for two nuclear proteins, one B cell restricted and the other ubiquitously occurring. Certain characteristics of the NF-W1 and NF-W2 pair recall the OTF-2/NF-A2 and OTF-1/NF-A1 pair that binds to the immunoglobulin octamer, but we demonstrate that the two protein pairs are distinguishable by several criteria. NF-W1 and NF-W2 interact differentially with their common GTTGCATC binding site, display a different affinity for it, and have molecular weights that differ by about 20,000. Yet, proteolysis experiments and cross-linking analyses indicate that the two W complexes show structural relatedness. PMID- 2564637 TI - cDNA sequence predicting an octapeptide-repeat antigen of Plasmodium falciparum. PMID- 2564636 TI - Involvement of proliferating cell nuclear antigen (cyclin) in DNA replication in living cells. AB - Proliferating cell nuclear antigen (PCNA) (also called cyclin) is known to stimulate the activity of DNA polymerase delta but not the other DNA polymerases in vitro. We injected a human autoimmune antibody against PCNA into unfertilized eggs of Xenopus laevis and examined the effects of this antibody on the replication of injected plasmid DNA as well as egg chromosomes. The anti-PCNA antibody inhibited plasmid replication by up to 67%, demonstrating that PCNA is involved in plasmid replication in living cells. This result further implies that DNA polymerase delta is necessary for plasmid replication in vivo. Anti-PCNA antibody alone did not block plasmid replication completely, but the residual replication was abolished by coinjection of a monoclonal antibody against DNA polymerase alpha. Anti-DNA polymerase alpha alone inhibited plasmid replication by 63%. Thus, DNA polymerase alpha is also required for plasmid replication in this system. In similar studies on the replication of egg chromosomes, the inhibition by anti-PCNA antibody was only 30%, while anti-DNA polymerase alpha antibody blocked 73% of replication. We concluded that the replication machineries of chromosomes and plasmid differ in their relative content of DNA polymerase delta. In addition, we obtained evidence through the use of phenylbutyl deoxyguanosine, an inhibitor of DNA polymerase alpha, that the structure of DNA polymerase alpha holoenzyme for chromosome replication is significantly different from that for plasmid replication. PMID- 2564638 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 15-1989. A 52-year-old man with neurofibromatosis and jaundice. PMID- 2564639 TI - Distal-less encodes a homoeodomain protein required for limb development in Drosophila. AB - The spatial organization of the Drosophila embryo depends on the activity of three axial pattern-forming systems. In addition to the anterior-posterior and dorsal-ventral systems that organize the segmented body plan, a proximal-distal pattern-forming system is required to provide positional information for the developing limbs. The development of both the larval and adult limbs depends directly on the activity of the Distal-less gene. Genetic analysis has shown that Distal-less functions as a developmental switch that is required to promote the development of limb structures above the evolutionary ground-state of body wall. Here we provide genetic evidence that indicates a graded requirement for Distal less activity during limb development. Reduction of this activity has a global effect on pattern formation in the limb. The molecular structure of the Distal less locus indicates that the gene encodes a homoeodomain-containing protein which is therefore likely to specify limb development through differential regulation of subordinate genes. PMID- 2564641 TI - Prescribing addictive medications. PMID- 2564640 TI - Temporally distinct pre- and post-synaptic mechanisms maintain long-term potentiation. AB - Long-term potentiation (LTP) in the hippocampus is widely studied as the mechanisms involved in its induction and maintenance are believed to underlie fundamental properties of learning and memory in vertebrates. Most synapses that exhibit LTP use an excitatory amino-acid neurotransmitter that acts on two types of receptor, the N-methyl-D-aspartate (NMDA) and quisqualate receptors. The quisqualate receptor mediates the fast synaptic response evoked by low-frequency stimulation, whereas the NMDA receptor system is activated transiently by tetanic stimulation, leading to the induction of LTP. The events responsible for maintaining LTP once it is established are not known. We now demonstrate that the sensitivity of CA1 neurons in hippocampal slices to ionophoretically-applied quisqualate receptor ligands slowly increases following the induction of LTP. This provides direct evidence for a functional post-synaptic change and suggests that pre-synaptic mechanisms also contribute, but in a temporally distinct manner, to the maintenance of LTP. PMID- 2564642 TI - [Children with autism and related contact disorders: medical aspects]. AB - In children with infantile autism or atypical pervasive developmental disorders somatic aspects play an important role. A review is presented of important hereditary, pre-, peri- and neonatal factors, findings at neurological examination, specific medical disorders and neurochemical and neurophysiological findings. Results of the medical examination of 15 children with autistic or atypical developmental disorders are presented. It is concluded that extensive medical examination of these children is indicated: in 8 out of 15 children a clinically relevant chromosomal, neurological or biochemical disorder could be detected. PMID- 2564643 TI - [Medical aspects of space flight]. PMID- 2564644 TI - [Diuretics in the treatment of arterial hypertension]. PMID- 2564645 TI - Better blocking agents? PMID- 2564647 TI - Taxol produces a predominantly sensory neuropathy. AB - Taxol, a plant alkaloid with promise as an antineoplastic agent, produced a predominantly sensory neuropathy in 16 of 60 patients treated in two phase I trials. This neuropathy occurred only at taxol doses greater than 200 mg/m2. Symptoms typically started 1 to 3 days following treatment, beginning in the hands and feet simultaneously in most patients. Electrophysiologic data suggests both axonal degeneration and demyelination. This previously undefined neurotoxic neuropathy most likely results from taxol's unique ability to produce microtubule aggregation in dorsal root ganglion cells, axons, and Schwann cells. PMID- 2564646 TI - Behavioural and electroencephalographic effects of excitatory amino acid antagonists and sigma opiate/phencyclidine-like compounds in rats. AB - The present work was a comparative study of the electrocortical and behavioural effects of phencyclidine-like drugs and sigma-opiate drugs and of some excitatory amino acid antagonists. Phencyclidine-like drugs and sigma-opiate drugs elicited three dose-dependent electrocortical patterns: 1. Increase of periods of cortical desynchronization 2. Increase of the amplitude of fast (20-30 Hz) cortical waves; 3. Appearance of typical cortical complexes consisting of slow-wave (0.5-2 Hz) sharp-wave complexes. These effects were accompanied by excitatory motor symptoms, such as stereotyped movements of the neck and limbs. The NMDA competitive antagonist D,L-2-amino-5-phosphonovaleric acid (D,L-AP5), administered intracerebroventricularly (0.25-2 mumol), elicited phencyclidine like stereotyped behaviour and cortical desynchronization, but failed to elicit/sigma typical cortical complexes. The non-selective amino acid antagonist cis-2,3 piperidine dicarboxilic acid (cis 2,3-PDA), administered intraperitoneally (3.3 mmol) failed to elicit both stereotyped behaviour and typical phencyclidine/sigma cortical complexes. The results strongly suggest the hypothesis that a reduction of excitatory amino acid neurotransmission, utilizing N-methyl-d-aspartate (NMDA)-preferring receptors, may be involved in the genesis of phencyclidine/sigma-induced stereotyped behaviour. PMID- 2564648 TI - Formation of long-term potentiation in superior colliculus slices from the guinea pig. AB - Superior colliculus slices of sagittal section were prepared from the guinea pig. Postsynaptic potential (PSP) was evoked in the superficial grey layer (SGL) after the electrical stimulation to optic layer. Tetanic stimulation to the optic layer elicited long-term potentiation (LTP) in the PSP of the SGL. Tetanic stimulation of 20 s in duration and 50 Hz in frequency was most effective for the formation of LTP. The LTP formation was masked during application of 2-amino-5 phosphonovalerate (APV), a specific antagonist for N-methyl-D-aspartate (NMDA) receptor, but LTP was observed when APV was removed from the perfusion medium. PMID- 2564649 TI - Involvement of locus coeruleus and noradrenergic neurotransmission in fentanyl induced muscular rigidity in the rat. AB - Whereas muscular rigidity is a well-known side effect that is associated with high-dose fentanyl anesthesia, a paucity of information exists with regard to its underlying mechanism(s). We investigated in this study the possible engagement of locus coeruleus of the pons in this phenomenon, using male Sprague-Dawley rats anesthetized with ketamine. Under proper control of respiration, body temperature and end-tidal CO2, intravenous administration of fentanyl (50 or 100 micrograms/kg) consistently promoted an increase in electromyographic activity recorded from the gastrocnemius and abdominal rectus muscles. Such an induced muscular rigidity by the narcotic agent was significantly antagonized or even reduced by prior electrolytic lesions of the locus coeruleus or pretreatment with the alpha-adrenoceptor blocker, prazosin. Microinjection of fentanyl (2.5 micrograms/50 nl) directly into this pontine nucleus, on the other hand, elicited discernible electromyographic excitation. It is speculated that the induction of muscular rigidity by fentanyl may involve the coerulospinal noradrenergic fibers to the spinal motoneurons. PMID- 2564650 TI - Glutamate immunoreactivity in rat dorsal root axons. AB - Approximately 8.5% of the unmyelinated and 2.5% of the myelinated primary afferent axons in lumbar dorsal roots of normal rats are immunostained for glutamate. Thus unmyelinated fibers are the predominantly immunostained population under the conditions of our experiments. The mean size of the unmyelinated fibers is greater at L6 than at L4 and L2. The meaning of this is not clear, but it may imply that pelvic visceral afferents are slightly larger than afferents from other areas. We emphasize that the immunostained axons can be demonstrated in otherwise normal animals, so changes in the percentages of labeled axons in response to various stimuli will be of interest. PMID- 2564651 TI - N-methyl-D-aspartate receptor activation is required for the induction of both early and late phases of long-term potentiation in rat hippocampal slices. AB - The possible involvement of N-methyl-D-aspartate (NMDA) receptors in mechanisms enabling the maintenance of long-term potentiation (LTP) was investigated in rat hippocampal slices. The action of the specific NMDA receptor antagonists (-)-2 amino-7-phosphonoheptanoic acid (D-APH) and 2-amino-5-phosphonovaleric acid (DL APV) as well as of the inactive isomer L-APH was tested on orthodromic population excitatory postsynaptic potential (EPSP) and population spike (PS) responses recorded extracellularly from CA1 pyramidal cells. If the active D-isomer of APH (10 microM) or DL-APV (50 microM), but not if L-APH was present during tetanization, both EPSP and spike potentiation were markedly reduced or even blocked for the whole recording period (8 h after tetanization). It is concluded that the NMDA receptor component expressed during tetanization is a necessary step not only for initiation but also for subsequent mechanisms enabling late phases of synaptic LTP. Some remaining potentiation of the population spike may be related to a second, NMDA-independent mechanism. PMID- 2564652 TI - Excitatory transmission in the dorsal horn is in part mediated through APV sensitive NMDA receptors. AB - The role of subtypes of excitatory amino acid receptor in synaptic transmission in the spinal dorsal horn has been studied in an in vitro slice preparation with well-preserved afferent inputs to the dorsal horn. Intracellular recordings were made from 30 dorsal horn neurons in laminae I-III of 18-28 day old rats. Superfusion of the slice with a Mg2+ zero solution resulted in an increase in the amplitude and duration of dorsal root-evoked excitatory postsynaptic potentials (EPSP) recorded intracellularly from dorsal horn neurons. Bath application of D-2 amino-5-phosphonovalerate (10(-5) M to 2.5 x 10(-5) M) or DL-2-amino-5 phosphonovalerate (10(-4) M to 2.5 x 10(-4) M) rapidly and reversibly abolished the later component of the EPSP evoked by activation of either group of primary afferents and selectively antagonized the N-methyl-D-aspartate-induced depolarization. PMID- 2564653 TI - Extracellular accumulation of glutamate in the hippocampus induced by ischemia is not calcium dependent--in vitro and in vivo evidence. AB - Calcium dependency of ischemia-induced increase in extracellular glutamate in the hippocampus was studied in vitro and in vivo. Perfusion of a low pO2 medium without glucose (in vitro ischemia) induced an increase in extracellular glutamate in rat hippocampal slices. This increase did not depend on Ca2+, which is in contrast with the observation that about 40% of membrane depolarization (50 mM KCl)-evoked release was Ca2+-dependent. In vivo cerebral ischemia of 5 min duration in gerbils also caused Ca2+-independent increase in extracellular glutamate in the hippocampus. The data suggest that the increase in extracellular glutamate induced by ischemia is not due to the enhanced release of neurotransmitter glutamate. PMID- 2564654 TI - Muscarinic inhibition of endogenous glutamate release from rat hippocampus synaptosomes. AB - The effects of acetylcholine (ACh) on the depolarization-evoked release of endogenous glutamic acid (Glu) have been studied using synaptosomes prepared from rat hippocampus and depolarized in superfusion with 15 mM KCl. Acetylcholine inhibited Glu release in a concentration-dependent way. The natural agonist was particularly effective causing 50% inhibition of Glu release at 10 microM in the absence of acetylcholinesterase (AChE) inhibitors. The inhibitory effect of ACh on the K+-evoked release of Glu was antagonized by the selective muscarinic receptor antagonist atropine but not by the nicotinic receptor antagonist mecamylamine. The data represent the first demonstration that muscarinic receptors located on Glu axon terminals in rat hippocampus may modulate the release of Glu. PMID- 2564655 TI - A comparison of nizatidine with the three other histamine receptor antagonists for duodenal ulcer therapy. AB - Nizatidine (Axid) is the newest of four histamine receptor antagonists approved by the Food and Drug Administration for treatment of duodenal ulcer and maintenance therapy. It is as effective as the other histamine blockers in healing active duodenal ulcers and in the prevention of relapse. Nizatidine is comparable in price to cimetidine (Tagamet). Nizatidine does not interfere with the hepatic metabolism of other drugs. No significant central nervous system side effects have been reported. It is safe to use in elderly patients or in patients with renal insufficiency. The dosage, however, should be decreased when renal function is impaired. PMID- 2564656 TI - Pseudonasolacrimal duct obstruction caused by nasal allergy. AB - While mechanical obstruction of the nasolacrimal duct is the most common cause of acquired unilateral epiphora, it is not the only one. Of the 94 patients evaluated with this symptom, 22 cases (23%) were caused by primary nasal changes in the form of nasal allergy. Many of the characteristic findings of mechanical obstruction were present. Several patients had previous nasolacrimal duct surgery, which failed to eliminate the epiphora. Nasal allergy was not initially considered either by previous examiners or the patients themselves, because of the absence of typical allergic symptoms. All of these patients, however, obtained relief from their unilateral tearing with treatment directed toward the nasal pathology only. Oral sympathomimetics and antihistamines were the most effective. Two patients improved with primary nasal surgery. PMID- 2564657 TI - Overexpression of erbB-2 or EGF receptor proteins present in early stage mammary carcinoma is detected simultaneously in matched primary tumors and regional metastases. AB - Membrane protein levels of erbB-2 and epidermal growth factor (EGF) receptor as well as gene aberrations affecting these proto-oncogenes in human mammary cancer were determined in primary and metastatic lesions. Among 57 patients erbB-2 gene amplification was detected in 11 tumors (19%). In 10 of these patients where expression levels could be assayed gene amplification was associated with a high level of erbB-2 protein. In contrast, EGF receptor gene amplification with over expression of the protein product was observed in 2 tumors (4%). In addition, 14 out of 53 (26%) primary tumors exhibited moderately increased erbB-2 protein levels in the absence of gene amplification. Similar aberrations resulting in overexpression of EGF receptor protein without detectable gene amplification were associated with 2 tumors (4%) among 47 patients analyzed. In 7 patients, expression level and gene copy numbers of erbB-2 or EGF receptor were similarly altered in the primary tumor and metastatic lesions derived from the same patient. Concordance of increased receptor gene expression in primary and metastatic lesions combined with the observation that such alterations are detectable as early as stage I and II mammary tumors, suggests that overexpression of erbB protooncogene family members can develop early in breast cancer and is maintained during tumor progression. Comparison of erbB-2 overexpression with clinical disease parameters revealed a correlation of this alteration with inflammatory mammary carcinoma (P = 0.042) implying an association of elevated erbB-2 protein levels with enhanced malignancy of the tumor cell in vivo. PMID- 2564658 TI - Design and development of pilus vaccines for Haemophilus influenzae diseases. PMID- 2564659 TI - Evaluation of pilus vaccines for prevention of experimental otitis media caused by nontypable Haemophilus influenzae. PMID- 2564660 TI - The surface of Branhamella catarrhalis: a systematic approach to the surface antigens of an emerging pathogen. AB - The incidence of B. catarrhalis as a cause of otitis media is increasing. As strategies for preventing infections caused by Streptococcus pneumoniae and nontypable H. influenzae are developed, the relative importance of B. catarrhalis will increase further in the next decade. A goal of studies of surface antigens is to identify potential vaccine components. Surface antigens of B. catarrhalis include OMPs, LOS and fimbriae. Preliminary studies indicate that OMPs and LOS are relatively antigenically conserved among strains. Future investigations should be directed toward identifying conserved antigens that will generate protective antibodies. PMID- 2564661 TI - Calf thymus DNA polymerase delta independent of proliferating cell nuclear antigen (PCNA). AB - DNA polymerase delta from calf thymus was purified under conditions that minimized proteolysis to a specific activity of 27,000 units/mg. The four step isolation procedure included phosphocellulose, hydroxyapatite, heparin-Sepharose and FPLC-MonoS. This enzyme consists of four polypeptides with Mr of 140, 125, 48 and 40 kilodaltons. Velocity gradient sedimentation in glycerol removed the 48 kDa polypeptide while the other three sedimented with the DNA polymerase activity. The biochemical properties of the three subunit enzyme and the copurification of 3'----5' exonuclease activity were typical for a bona fide DNA polymerase delta. Tryptic peptide analysis showed that the 140 kDa polypeptide was different from the catalytic 180 kDa polypeptide of calf thymus DNA polymerase alpha. Both high Mr polypeptides (140 and 125 kDa) were catalytically active as analysed in an activity gel. Four templates were used by DNA polymerase delta with different preferences, namely poly(dA)/oligo(dT)12-18 much much greater than activated DNA greater than poly(dA-dT) greater than primed single stranded M13DNA. Calf thymus proliferating cell nuclear antigen (PCNA) could not stimulated this DNA polymerase delta in any step of the isolation procedure. If tested on poly(dA)/oligo(dT)12-18 (base ratio 10:1), PCNA had no stimulatory effect on DNA polymerase delta when tested with low enzyme DNA ratio nor did it change the kinetic behaviour of the enzyme. DNA polymerase delta itself did not contain PCNA. The enzyme had an intrinsic processivity of several thousand bases, when tested either on the homopolymer poly(dA)/oligo(dT)12-18 (base ratio 64:1) or on primed single-stranded M13DNA. Contrary to DNA polymerase alpha, no pausing sites were seen with DNA polymerase delta. Under optimal in vitro replication conditions the enzyme could convert primed single-stranded circular M13 DNA of 7,200 bases to its double-stranded form in less than 10 min. This supports that a PCNA independent DNA polymerase delta exists in calf thymus in addition to a PCNA dependent enzyme (Lee, M.Y.W.T. et al. (1984) Biochemistry 23, 1906-1913). PMID- 2564663 TI - PstI RFLP for the anonymous [D13S2] (p9D11) probe. PMID- 2564662 TI - Expression of Hox-2.4 homeobox gene directed by proviral insertion in a myeloid leukemia. AB - The presence of an altered Hox-2.4 gene in the WEHI3B murine myeloid leukemia suggests that homeobox genes may contribute to neoplasia. A survey of 31 leukemia cell lines of the myeloid, lymphoid and erythroid lineages revealed that Hox-2.4 was expressed only in WEHI3B and the pre-B lymphoid line 70Z/3, in which no DNA rearrangement was observed. To clarify the WEHI3B alteration and normal Hox-2.4 structure, we have sequenced near full length cDNA clones from WEHI3B and 70Z/3, and the 5' portion of the normal Hox-2.4 gene. A WEHI3B cDNA clone demonstrates that an intracisternal A-particle (IAP) provirus has inserted within the first exon of the gene and generated a Hox-2.4 mRNA with a 5' sequence derived from the IAP long terminal repeat. A remarkable degree of similarity found between the amino acid sequences of Hox-2.4 and Hox-3.1, which reside on different chromosomes, supports the notion that an ancient homeobox gene cluster has been duplicated and dispersed early in vertebrate evolution. PMID- 2564664 TI - BANI detects a biallelic DNA polymorphism of the human phenylethanolamine N methyltransferase [hPNMT] gene. PMID- 2564665 TI - New TaqI RFLPs at the DXS52 (St14) locus in the black population. PMID- 2564666 TI - ApaI dimorphism at the human vitamin D receptor gene locus. PMID- 2564667 TI - PstI polymorphism of the alpha 1-antitrypsin-like gene. PMID- 2564668 TI - Two 47z [DXYS5] RFLPs on the X and the Y chromosome. PMID- 2564669 TI - Previously unreported NcoI RFLP for human CSF1R. PMID- 2564670 TI - SstI polymorphism revealed by anonymous probe cpl 2.6 [D5S89] which maps to 5q21- --5q31. PMID- 2564671 TI - RFLPs detected at 2 pter-p12 with a thyroid peroxidase cDNA probe, TPO3 (McKusick no. 27450). PMID- 2564672 TI - Cancer pain control: one state's experience. AB - The World Health Organization estimates that approximately 3.5 million people suffer daily from cancer pain. Approximately 30% to 40% of people with intermediate stages of cancer and 55% to 90% of patients with advanced or terminal disease have pain. A multidisciplinary effort is under way in Wisconsin to improve cancer pain management. In recognition of the state's initiative, the World Health Organization has designated Wisconsin a demonstration state for cancer pain management. Support for staffing is provided by the United States Public Health Service Interagency Committee on Pain and Analgesia. In December 1986, 65 people representing 50 health organizations met to develop a comprehensive plan of action to improve pain management in people with cancer. In this article the various statewide activities focused at improving cancer pain management are discussed. PMID- 2564673 TI - Control of pain in cancer patients. AB - Almost three quarters of patients with cancer have severe pain, from invasion of the cancer itself, from effects of therapy, or from causes unrelated to the cancer (but often exacerbated by it). With the proper pain-management strategy, however, pain can be controlled in most patients. The analgesic ladder for pain control, promoted by the World Health Organization, begins with a nonnarcotic agent, progresses to a weak narcotic plus a nonnarcotic, and finally reaches a strong narcotic. Adjuvant agents, which increase the analgesic potency of the drug being used, may be added at any level. The most common reasons for inadequate pain control in cancer patients are incorrect narcotic dosing and incorrect switching from one narcotic to another and from one route of administration to another. Factors that influence pain management (eg, fear, anxiety, sleep disturbance) should be treated as well with appropriate medications, behavioral therapy, counseling, hypnosis, and other supportive techniques. These points are illustrated in the case report (see box, page 328). PMID- 2564674 TI - Fimbriae of Bacteroides nodosus: protein engineering of the structural subunit for the production of an exogenous peptide. AB - The pattern of sequence variation between Bacteroides nodosus fimbrial subunits of different serotypes suggests a degree of flexibility, which might be exploited for protein engineering approaches for the expression of other peptides. We have tested this using the well-characterized peptide epitope from VP1 of foot-and mouth disease virus (FMDV), residues 144-159: LRGDLQVLAQKVARTL (strain 01-BFS). Using bacterial codon usage, several oligonucleotides were designed for the substitution of this sequence internally at hypervariable regions of the fimbrial subunit (aligned for maximum homology), and for its addition at the carboxyterminus with a diglycine spacer as a flexible hinge. Following site directed mutagenesis in phage M13, the modified genes were placed under PL promoter control and placed in a broad host range vector. Analysis of the variant proteins expressed in E. coli showed that these substitutions affected, to varying extents, recognition by both anti-fimbrial and anti-FMDV antibodies, indicating that hypervariable region 2 is a major antigenic determinant of the fimbrial subunit and that local stereochemical effects can influence antibody binding to the FMDV peptide antigenic determinant. In Pseudomonas aeruginosa, viable transformants could only be obtained with the mutant gene encoding the carboxy-terminal graft. These cells provided fimbrial preparations comprised of the modified subunit. This then constitutes the prototype for the development of a general expression system for the production of vaccine epitopes and other bioactive peptides. Furthermore, there is considerable scope for further modification of the system, for example by engineering specific chemical or protease cleavage sites for release of the grafted peptide. Alternatively, the fimbriae themselves may serve as a useful supramolecular carrier or adjuvant for immune provocation. PMID- 2564675 TI - Amino acid substitutions in albumin variants found in Brazil. AB - Conventional horizontal starch-gel electrophoresis in four buffer systems and structural studies were performed on four albumin variants, and the findings were compared with similar previous data. Albumins Coari I and Porto Alegre I have a previously unreported amino acid substitution (glutamic acid replaced by lysine at position 358, denoted 358 Glu----Lys). The alteration in albumin Porto Alegre II (501 Glu----Lys) is the same as that found for three alloalbumins of Asiatic origin, designated Vancouver, Birmingham, and Adana. Albumin Oriximina I has the same exchange as albumin Maku (541 Lys----Glu). Some of these findings can be explained only by the occurrence of independent mutations at the same site in the albumin gene. They also point to a third cluster of mutations in that gene, indicating hypermutability in some of its segments. PMID- 2564676 TI - Formation of pilin in Pseudomonas aeruginosa requires the alternative sigma factor (RpoN) of RNA polymerase. AB - The promoter region of the Pseudomonas aeruginosa pilin gene has a high degree of similarity to the nitrogen-regulated promoters of enteric bacteria. These promoters are recognized by the alternative sigma factor of RNA polymerase, termed RpoN (NtrA or GlnF). This observation suggested that the P. aeruginosa pilin gene may be transcribed by the RpoN-containing RNA polymerase. We, therefore, cloned the RpoN gene from P. aeruginosa into Escherichia coli (where it formed a functional product) and used that cloned gene to construct a mutant of P. aeruginosa that was insertionally inactivated in its RpoN gene. This mutant failed to synthesize pilin, indicating that the RpoN sigma factor is required for transcription of the pilin gene. PMID- 2564677 TI - CD4+ lymphocyte function with early human immunodeficiency virus infection. AB - The pathogenesis of cellular immune deficiency following human immunodeficiency virus (HIV) infection could result from quantitative and/or qualitative dysfunction of the CD4+ lymphocyte population. To better characterize the T-cell response to soluble antigen with HIV infection, we have isolated peripheral blood lymphocytes and purified populations of CD4+ lymphocytes from healthy HIV antibody-positive subjects, patients with acquired immunodeficiency syndrome (AIDS)-related complex (ARC), and healthy HIV antibody-negative controls. T lymphocyte function was determined by proliferative response to lectin (phytohemagglutinin), phorbol 12-myristate 13-acetate (PMA), calcium ionophore, purified recombinant HIV envelope gp120, tetanus toxoid antigen, and tetanus toxoid antigen in the presence of recombinant gp120 or purified recombinant soluble CD4. PBLs and CD4+ lymphocytes from asymptomatic HIV-infected subjects responded equally well to lectin, PMA, and/or calcium ionophore and to tetanus toxoid as cells from uninfected control subjects. The cells that proliferated in response to a soluble antigenic stimulus did not respond to gp120. Cells from subjects with ARC had a selective antigen recognition defect independent of the number of CD4+ lymphocytes. Recombinant gp120 inhibited CD4+ lymphocyte proliferation to antigenic stimulus by 30-40%. Recombinant soluble CD4, a proposed therapeutic for HIV, had no effect on T-cell response to antigen. A selective antigen recognition response was not compromised early in HIV infection but was compromised in subjects with ARC. Inhibition of proliferation to tetanus toxoid by gp120 suggests that HIV may affect major histocompatibility complex II restricted antigen recognition independent of CD4+ cell loss. PMID- 2564678 TI - Somatostatin analogues inhibit growth of pancreatic cancer by stimulating tyrosine phosphatase. AB - Several analogues of somatostatin were examined in the Mia PaCa-2 human pancreatic cancer cell line for their ability to promote tyrosine phosphatase activity affecting the receptors for the epidermal growth factor. The inhibition of growth of the Mia PaCa-2 cells in culture was also evaluated to determine the mechanism of action of somatostatin analogues and their relative effectiveness in inhibiting cancer growth. Of the analogues tested D-Phe-Cys-Tyr-D-Trp-Lys-Val-Cys Trp-NH2 (RC-160) caused the greatest stimulation of tyrosine phosphatase activity. Analogue D-Phe-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH2 (RC-121) had less effect but was more potent than somatostatin-14. Analogue D-Phe-Cys-Phe-D-Trp-Lys Thr-Cys-Thr(ol) (SMS 201-995) produced no significant dephosphorylation. The analogues displayed the same order of activity in assays on growth inhibition of Mia PaCa-2 cells in cultures. Analogue (SMS-201-995) caused virtually no tyrosine phosphatase stimulation or growth inhibition in this cancer cell line, although it possesses a much higher antisecretory activity than somatostatin-14 in normal tissues. These observations indicate that somatostatin and some of its analogues can act as growth inhibitors in cancer cells through the activation of tyrosine phosphatase. These data reinforce the view that somatostatin analogue RC-160 and related compounds could be used for treatment of pancreatic cancer. PMID- 2564681 TI - Increased uptake of methylated low-density lipoprotein induced by noradrenaline in carotid arteries of anaesthetized rabbits. AB - Atherosclerosis is accelerated in hyperlipidaemias but, apart from the concentration of low-density lipoprotein (LDL) in the blood, very little is known about other influences on the disease process. We now provide evidence that in anaesthetized rabbits the atherogenic uptake of LDL by arterial walls is accelerated by noradrenaline at its physiological concentrations in rabbit and human blood. The principle of the experiments was to compare the uptake of intravenously injected, radioactively labelled LDL, methylated to prevent removal by high-affinity receptors, in the two carotid arteries of anaesthetized rabbits after infusing low concentrations of noradrenaline into one carotid and saline as control into the other, the volume rates of infusion being about 1% of the carotid blood flows. Human LDL, which behaves sufficiently like rabbit LDL for these purposes, was prepared, methylated and radio-iodinated by standard methods. At the end of the infusions, the arteries were excised and their radioactivities determined. Noradrenaline infused for 2 h to produce local blood concentrations of nominally 1, 10, 50 and 100 nM significantly increased the LDL radioactivities of the walls of the noradrenaline-infused carotids. Concentrations of nominally 100 nM also increased the LDL radioactivities of the walls of the saline-infused carotids; this was associated with significant increases in their blood noradrenaline concentrations. These results may contribute towards an explanation for the accelerated atherosclerosis and the increased incidence of its clinical manifestations in conditions associated with elevated blood noradrenaline concentrations, including the episodic increases associated with stress and cigarette smoking as well as the more persistent increases caused by phaeochromocytoma. PMID- 2564679 TI - In vitro transcription directed from the somatostatin promoter is dependent upon a purified 43-kDa DNA-binding protein. AB - In vitro transcription analyses were used to establish the biological function of a 43-kDa affinity-purified DNA-binding protein. The 43-kDa affinity-purified protein protects the region from position -59 to position -35 of the somatostatin promoter from DNase I digestion. This region of the somatostatin promoter harbors the TGACGTCA motif, also found and required for function in a number of other cAMP-responsive and adenovirus E1A-inducible promoters. Efficient and authentic transcription in vitro directed from the somatostatin promoter requires the TGACGTCA promoter element. In vitro transcription assays performed in the presence of somatostatin (positions -60 to -29), enkephalin (positions -105 to 71), and adenovirus type 5 E3 gene (positions -72 to -42) competitor fragments, harboring similar TGACGTCA motifs, selectively inhibit transcription directed from the somatostatin promoter, suggesting that the TGACGTCA element is the site of interaction of a somatostatin gene transactivator. Furthermore, extracts depleted of the TGACGTCA-binding activities by affinity chromatography utilizing a biotinylated oligonucleotide-avidin resin, are incapable of directing transcription from the somatostatin but not from the adenovirus major late promoter. Addition of the purified 43-kDa protein to the affinity-depleted extract restores transcription from the somatostatin promoter. These results are consistent with the 43-kDa protein being a trans-activator of the somatostatin gene. PMID- 2564682 TI - Growth and elimination of nerve terminals at synaptic sites during polyneuronal innervation of muscle cells: a trophic hypothesis. AB - This paper examines the possibility that the elimination of synapses from cells arises from a competition between the nerve terminals for trophic molecules made available by the cells. This idea is applied to the elimination of synapses that occurs during the polyneuronal innervation of muscle cells which accompanies both the development and reinnervation of muscles. In the proposed model, each motorneuron makes the same amount of receptor in its soma for a trophic molecule provided in limited quantities by each muscle cell; this receptor is then distributed to the collateral terminals of the motorneuron in concentrations proportional to the amount of receptor made in the soma by the motorneuron; the more collateral terminals initially possessed by a motorneuron the less will be their concentration of receptor. The receptors in the several collateral terminals on a muscle cell then compete for the trophic molecule provided by the muscle, and terminal growth is proportional to the number of receptor-trophic molecule bonds formed. An autocatalytic effect has been introduced whereby the increase in size of a terminal accelerates the rate by which the trophic molecule is made available to that terminal for bonding with its receptors. In addition, the affinity between nerve terminal receptors and muscle molecules can be varied in the model. Finally, motorneuron cell death has been analysed as the elimination of neurons that have insufficient terminal area to take up a growth factor in amounts that will allow for the survival of the neuron. PMID- 2564680 TI - Cachectin alters anterior pituitary hormone release by a direct action in vitro. AB - Cachectin (tumor necrosis factor) is a powerful macrophage hormone released during infection, which circulates in blood to produce diverse effects in the organism. We examined the effect of cachectin on release of anterior pituitary hormones from either hemipituitaries or dispersed pituitary cells incubated in vitro. The action of cachectin on dispersed cells was demonstrable only after 2 hr of incubation. With this incubation time, the protein produced a dose-related stimulation of release of adrenocorticotropin (ACTH), growth hormone (GH), and thyrotropin (TSH), but not of prolactin (Prl), from both hemipituitaries and dispersed cells. The doses required for stimulation were low in the case of hemipituitaries, usually of the order of 10(-12) M, whereas they were higher by one or two orders of magnitude with the dispersed pituitary cells. This may be related either to loss of receptors for the protein during the dispersion procedure or to the fact that in the hemipituitary system cell interactions are facilitated because the cells are close to each other. In the dispersed cell system cachectin evoked a dose-related decrease in cyclic AMP content. Incubation with somatostatin lowered the cyclic AMP content of the cells and depressed GH output without altering output of TSH or Prl. When somatostatin and cachectin were incubated together with the cells, the suppression of cyclic AMP production was abolished; TSH and Prl release were stimulated, but the action of cachectin to stimulate GH release was blocked. The stimulation of Prl release by cachectin in the presence of somatostatin may be related to the elevation of cyclic AMP, a known stimulator of Prl release. The cyclooxygenase inhibitor indomethacin nearly completely blocked the stimulatory effect of cachectin on release of GH and TSH from dispersed pituitary cells but had only a slight and nonsignificant attenuating effect on its ACTH-releasing action. These results suggest that at least part of the stimulatory action of the peptide on pituitary hormone release is brought about by prostaglandins. The failure of indomethacin to block the release of ACTH induced by cachectin suggests that other mechanisms may be involved in the release of ACTH induced by this peptide. Since the concentrations of cachectin required to stimulate pituitary hormone release are similar to those that are encountered in plasma during infection, it is likely that this direct pituitary action has pathophysiological significance. PMID- 2564684 TI - Amplifiers and the handicap principle in sexual selection: a different emphasis. AB - Population genetic models have shown that female choice is a potential cause of the evolution of male display. In these models the display is assumed to be the immediate object of female choice. Here I present an explicit genetic model that shows that male display can evolve as a consequence of female choice even when the display is not the immediate object of choice. When females initially base their preferences on the existence of variance in a cue that is correlated with male viability, a rare display can evolve to fixation if it amplifies the previously recognized differences in males, (i.e. if it increases the resolution power of females with respect to the original cue). By definition, amplifying displays (or amplifiers) increase mating success of the more viable males and decrease mating success of the less viable males. Therefore, the higher the frequency of the preferred, more viable males, the more likely it is that amplifiers will evolve to fixation. The evolution of an amplifier is further facilitated by a genetic association that is built up between the amplifier allele and the more viable allele. If the expression of the amplifier is limited to the more viable males, the amplifier will evolve to fixation provided only that the change in total fitness to the more viable males (higher mating success, lower viability), is positive. PMID- 2564683 TI - Restoration of fast muscle characteristics following cessation of chronic stimulation: physiological, histochemical and metabolic changes during slow-to fast transformation. AB - Implantable electronic stimulators were used to subject fast-twitch tibialis anterior and extensor digitorum longus muscles of adult rabbits to a chronically increased level of use. Stimulation was discontinued after 6 weeks and physiological, histochemical and biochemical properties of the muscles were examined at intervals over the ensuing 20 weeks. Previous work had shown that 6 weeks of stimulation was sufficient to bring about a substantial transformation of type in fast-twitch muscles, which then exhibited much of the character of muscles of the slow-twitch type. The present experiments showed that these stimulation-induced changes were completely reversible. The time-course of reversion was such that the muscles had recovered their original fast properties by about 12 weeks after the cessation of stimulation. The contractile characteristics and post-tetanic potentiation typical of fast muscle returned rapidly, in only 3-4 weeks, and over the same period the proportion of histochemical type 1 fibres declined from about 70% to control levels. Changes in fatigue-resistance, capillary density and enzyme activity followed a more prolonged time-course; in particular, the decline in the activity of enzymes of oxidative metabolism corresponded closely to that already established for the mitochondrial volume fraction. Reacquisition of fast properties was not accompanied by any changes in specific force-generating capacity. Observations from these experiments and from a related morphological study fit into a 'first in, last-out' pattern for the response to stimulation and recovery. The slow-to fast reversion that takes place during the recovery period provides a further opportunity for testing causal associations within the events underlying type transformation. It has important consequences for therapeutic applications that make use of the fatigue-resistant character of chronically stimulated muscle. PMID- 2564685 TI - Possible role of bacterial adherence and bacterial adhesins in sepsis and septic shock. PMID- 2564686 TI - The neuropathic foot--a management scheme: a case report. AB - The purpose of this case report is to present a management approach that was effective in the healing and long-term care of a neuropathic plantar ulcer in a patient with diabetes mellitus. The report demonstrates that a successful management program must go beyond the stage of healing and include patient education and techniques for reducing plantar pressures to prevent the recurrence of plantar ulcers. PMID- 2564688 TI - A neurotransmitter basis for Eysenck's theory of personality. AB - A physiological basis for Eysenck's theory of personality is proposed which is different from those bases suggested by Eysenck himself. Psychoticism, neuroticism and extraversion are associated with varying levels of dopamine, serotonin, and norepinephrine, respectively. PMID- 2564687 TI - Endogenous sleep-promoting substances and sleep regulation. PMID- 2564689 TI - Benzodiazepines in the treatment of alcoholism. AB - This chapter comprises three sections that cover the main aspects of benzodiazepines and alcohol: (1) the basic pharmacology of benzodiazepines; (2) use of benzodiazepines in the treatment of withdrawal; and (3) the use of benzodiazepines in treating alcoholics. The basic studies suggest that a major site of action of alcohol may be the GABA/benzodiazepine receptor complex and that compensatory alterations in this complex may underly withdrawal. In the section on alcohol withdrawal, interactions between the GABA/benzodiazepine receptor complex, sympathetic nervous system, and hypothalamic-pituitary-adrenal axis are discussed. Use of benzodiazepines in the treatment of the alcohol withdrawal syndrome are reviewed, including the possibility that the benzodiazepines may prevent withdrawal-induced "kindling." Lastly, we review indications for, and efficacy of, benzodiazepines in long-term treatment of patients with alcoholism. Benzodiazepines are not indicated for the treatment of alcoholism. Furthermore, they have very few indications in alcoholics and their dependency-producing potency has to be appreciated when they are used in patients with alcoholism. PMID- 2564690 TI - [Mucoviscidosis: current diagnostic possibilities. Applications in perinatology]. AB - The gene of cystic fibrosis is localised on the long arm of chromosome 7. DNA probes placed close to the gene enable a study of restriction polymorphism to follow the transmission of the gene in index families. It is now possible to counsel those families, who already have an affected child, with an early antenatal diagnosis at ten weeks after the last period. In our personal experience, based on a study of the genotype of 48 families, 70% were informative when they were studied by two probes corresponding to the local pJ3.11 and met. When the latter probes Km19-XV2c were studied concurrently useful information was achieved in 96%. DNA analysis non enables the detection of the chromosome carrying the deleterious gene in practically every family where there is a child suffering from the disease. PMID- 2564691 TI - [Treatment with anti-epileptic drugs--a survey of mentally handicapped adults]. AB - Type and frequency of anticonvulsive drug treatment have been analyzed in the case of 1154 mentally retarded adults. Compared with international studies, anticonvulsive drug prevalence is low and amounts to 23.0% in the institutionalized group and 11.9% in the noninstitutionalized. Anticonvulsive drug prevalence decreases significantly with increasing age. A comparison of the 1986 anticonvulsive medication with that of 1980 shows distinctly rarer prescription of phenytoin and phenobarbital, and a decrease in the treatment with three or more different anticonvulsants in favour of treatment with only one or two anticonvulsive drugs in 1986. PMID- 2564692 TI - [Pillars of therapy of chronic obstructive bronchitis]. AB - While bronchial asthma is defined on the basis of pulmonary function, emphysema has a patho-anatomic substrate. Chronic bronchitis on the other hand is defined clinically: productive cough of three months duration per year occurring in two successive years. Complete abstinence of smoking is a prerequisite for a meaningful anti-obstructive therapy. Beta-2-adrenergic stimulation and corticosteroids represent the treatment of choice. Beta-2-agonists are effectively complemented by anticholinergic drugs. Oral or parenteral corticosteroids should be used liberally for exacerbations. For prolonged treatment topical steroids are prescribed exclusively in order to prevent systemic side effects. The type of inhalation therapy (electric nebulizer, aerosol spray, insufflator) has to be adapted to the individual patient. Correct application of inhalation-technique is important. Topical steroids should be administered by means of a spacer chamber. Physical therapy is the most important adjuvant treatment. Selective respiration-training, a correction of breath technique and autogenic bronchial drainage manoeuvres have to be learned in the setting of a rehabilitation program and are controlled frequently by the treating physician. The integration of all these measures into a treatment and rehabilitation plan is facilitated in a specialized clinic. PMID- 2564693 TI - Toxic shock syndrome toxin 1 is encoded by a variable genetic element. AB - The primary cause of toxic shock syndrome is toxic shock syndrome toxin 1 (TSST 1), a 22,049-dalton exotoxin. Approximately 20% of Staphylococcus aureus isolates produce TSST-1; the production of this toxin is therefore a variable genetic trait. The TSST-1 gene and its flanking sequences are found on a genetic element that is present in TSST-1-positive isolates and absent in TSST-1-negative isolates. Preliminary sequence data and Southern hybridization experiments with the cloned flanking sequences have provided evidence that the TSST-1 element is 4 7 kilobases in size. Hybridization analysis of whole-cell DNA from two genetically mapped TSST-1-positive strains has demonstrated that the TSST-1 element has at least two chromosomal locations. This finding suggests that the element is mobile. Biotyping of 75 TSST-1-positive isolates showed that the large majority were tryptophan-negative, and Southern hybridization analysis of whole cell DNA from these isolates revealed a common blotting pattern--an observation suggesting that these strains are clonal. PMID- 2564694 TI - Neuropeptide Y, peptide YY, and sympathetic control of rectal tone and anal canal pressure in the cat. AB - Sympathetic mechanisms in the regulation of rectal tone and anal canal pressure have been investigated in the cat. Immunohistochemical studies demonstrated neuropeptide Y-like immunoreactivity in noradrenergic cell bodies of the inferior mesenteric ganglion, in nerve endings in the circular smooth-muscle layers of the rectum and internal anal sphincter, in the myenteric plexuses, and around blood vessels and anal glands. Peptide YY-like immunoreactivity was demonstrated exclusively in endocrine cells of the rectal mucosa. By radioimmunoassay the amount of neuropeptide Y in the inferior mesenteric ganglion was found to be 28.5 (17.8-66.6) pmol g-1, in the rectum 0.8 (0.7-1.6) pmol g-1, and in the anal canal region 0.8 (0.5-1.0) pmol g-1 of tissue. Neuropeptide Y (4-80 pmol kg-1 min-1 intravenously) and peptide YY (1-25 pmol kg-1 min-1 intravenously) increased rectal tone and anal canal pressure. The effects were not inhibited by guanethidine, phentolamine, or propranolol. Noradrenaline (100-1000 pmol kg-1 min 1 intravenously) increased rectal tone and anal canal pressure. These effects were blocked by phentolamine and propranolol. Electric stimulation of the lumbar colonic nerves and the hypogastric nerves (8 Hz) increased rectal tone and anal canal pressure. After propranolol and phentolamine the responses partly remained. On lumbar colonic nerve stimulation the remaining responses of the rectum and anal canal were 60% (40-68%) (p less than 0.05) and 20% (15-29%), respectively, whereas on hypogastric stimulation the remaining rectal and anal responses were 32% (20-42%) (p less than 0.05) and 31% (20-47%) (p less than 0.05). Further administration of guanethidine abolished the remaining responses of the rectum and anal canal. Since neuropeptide Y is present in sympathetic neurons and mimics non-adrenergic responses to nerve stimulation, this peptide is a transmitter candidate for the effects resistant to adrenergic blocking agents but sensitive to guanethidine. Peptide YY may also play a role in sphincter control, but it cannot be excluded that it activates mechanisms similar to those activated by neuropeptide Y. PMID- 2564695 TI - [An unusual cause of gangrene: cold injury caused by liquid nitrogen]. AB - A case is described of gangrene of the foot and lower leg due to cold injury following exposure to liquid nitrogen. The severe damage to the entire microcirculation was followed by irreversible ischemia finally requiring amputation of the lower leg. Histology revealed conspicuous lesions of the small vessels within the entire soft tissues, with subsequent thrombotic occlusion and circulatory blockade. Some recommendations concerning occupational exposure to liquid nitrogen are added. PMID- 2564696 TI - Multidrug resistance in cancer. AB - Chemotherapy often fails because a tumor develops resistance to an array of different drugs. A single glycoprotein turns out to be responsible: it proliferates in some cells and pumps out the drugs. Now that the protein pump has been identified it may be possible to interfere with its action or to make it the target for drugs that destroy the cancer cell. PMID- 2564697 TI - Astrocytes. PMID- 2564698 TI - Cnidocyte mechanoreceptors are tuned to the movements of swimming prey by chemoreceptors. AB - Cnidocytes, the stinging cells of cnidarians, discharge nematocysts in response to physical contact accompanied by the stimulation of specific chemoreceptors. Cnidocytes in fishing tentacles of a sea anemone are now found to discharge nematocysts preferentially into targets vibrating at 30, 55, and 65 to 75 hertz. Moreover, in the presence of submicromolar concentrations of known chemosensitizers, such as N-acetylated sugars and mucin, these optima shift to 5, 15, 30, and 40 hertz, frequencies that correspond to the movements of swimming prey. Hence, chemoreceptors for these substances tune cnidocyte mechanoreceptors to frequencies that match the movements of the prey. PMID- 2564699 TI - Commitment of neural crest cells to the sensory neuron lineage. AB - Clonal cultures and monoclonal antibodies against a lineage-specific epitope, stage-specific embryonic antigen-1 (SSEA-1) were used to analyze the commitment of quail neural crest cells to the sensory neuron pathway. There were two distinct populations of sensory cells at the time of gangliogenesis. Postmitotic neuroblasts that remained in close association with the neural tube coexisted with a large number of pluripotent cells that formed the leading edge of the emigrating cells and gave rise to sensory and autonomic neuroblasts and to melanocytes. The data suggest a dual origin of spinal sensory neuroblasts and a predominantly late divergence of the autonomic and sensory lineages. PMID- 2564700 TI - Possible role of carbamates in neurotoxicity and neurotransmitter inactivation. PMID- 2564701 TI - In vivo modulation of cytolytic activity and Thy-1 expression in TCR-gamma delta+ intraepithelial lymphocytes. AB - Although the functional aspects of the alpha beta T cell antigen receptor (TCR) found on most peripheral T cells are well described, the function of the gamma delta TCR remains unclear. Murine intraepithelial lymphocytes (IEL) of the small intestine are CD8+, express the gamma delta TCR, and are constitutively lytic. Fresh IEL from germ-free mice had no lytic activity. Moreover, whereas IEL from normal mice are 30 to 50 percent Thy-1+, IEL from germ-free did not express Thy 1. Acclimation of germ-free mice to nonsterile conditions resulted in the generation of Thy-1+ IEL and induction of lytic activity. Thus CD8+ TCR-gamma delta IEL were regulated by externally derived stimuli via a specific functional interaction between IEL and gut-associated antigens. PMID- 2564702 TI - Defensive behaviors in infant rhesus monkeys: environmental cues and neurochemical regulation. AB - To survive, primates must detect danger in time to activate appropriate defensive behaviors. In this study, the defensive behaviors of infant rhesus monkeys exposed to humans were characterized. It was observed that the direction of the human's gaze is a potent cue for the infant. Infants separated from their mothers were active and emitted frequent distress vocalizations. When a human entered the room but did not look at the infant, it became silent and froze in one position. If the human stared at the infant, it responded with aggressive barking. Alterations of the opiate system affected the frequency of the infant's distress calls without affecting barking and freezing, whereas benzodiazepine administration selectively reduced barking and freezing. This suggests that opiate and benzodiazepine systems regulate specific defensive behaviors in primates and that these systems work together to mediate behavioral responses important for survival. PMID- 2564703 TI - Factor XIIIa-expressing dermal dendrocytes in AIDS-associated cutaneous Kaposi's sarcomas. PMID- 2564704 TI - Effects of nicotinic agonists and antagonists on N-methyl-D-aspartate-induced 3H norepinephrine release and 3H-(1-[1-(2-thienyl)cyclohexyl]-piperidine) binding in rat hippocampus. AB - The nicotinic agonists dimethylphenylpiperazinium iodide (DMPP) and carbachol (CARB) as well as (-)nicotine [-)NIC) were tested alone and in combination with N methyl-D-aspartate (NMDA) for their abilities to enhance the efflux of 3H norepinephrine (NE) from slices of rat hippocampus. CARB and (-)NIC produced small, transient increases in NE efflux, while DMPP produced larger, longlasting increases. Inasmuch as the nicotinic antagonists mecamylamine (MECA) and hexamethonium (C6) did not consistently inhibit the increases in NE efflux produced by these agonists, the role of a nicotinic receptor in mediating these responses is uncertain. CARB and DMPP enhanced the ability of NMDA to stimulate NE release, while (-)NIC did not. MECA, but not C6, was found to selectively antagonize NMDA-stimulated NE release that did not appear to involve a nicotinic receptor. Binding studies indicated that MECA and the related nicotinic antagonist pempidine produced an inhibition of NMDA-stimulated NE release by an action at the PCP receptor that is known to be linked to the NMDA receptor ionophore complex. These data suggest that the actions of these ganglionic blocking agents on excitatory responses in the hippocampus involve inhibition of excitatory amino acid as well as nicotinic receptors. PMID- 2564705 TI - Is current treatment increasing asthma mortality and morbidity? PMID- 2564706 TI - Alterations of guinea pig alveolar macrophage oxidative metabolism by nicotine. AB - The influence of nicotine, a cholinergic agonist, on guinea pig pulmonary alveolar macrophage (PAM) oxidative metabolism was examined. Nicotine caused a concentration-dependent enhancement or inhibition of chemiluminescence response and superoxide anion release by zymosan-stimulated PAM. Thus, at 5 x 10(-10) and 5 x 10(-8) M of nicotine the chemiluminescence response was augmented to 132 and 113%, respectively. At higher concentrations, such as 5 x 10(-7) and 1 x 10(-4) M, however, these responses were inhibited to 83 and 51% of the control, respectively. Similarly, at 5 x 10(-10) and 5 x 10(-9) M nicotine, superoxide anion release was enhanced to 226 and 209% of the control, respectively. Higher concentrations of nicotine, 5 x 10(-5) and 5 x 10(-4) M, inhibited this response to 53 and 58% of the control, respectively. Neither the potentiating nor the inhibitory effect of nicotine was affected by a muscarinic (atropine) or nicotinic (hexamethonium) cholinergic antagonist. None of the drugs examined, by themselves, stimulated PAM oxidative metabolism or influenced oxyradical generation by a cell-free system. This study demonstrates that nicotine, a major component of cigarette smoke, may play a significant role in the pathogenesis of some pulmonary diseases. PMID- 2564707 TI - Hepaticojejunostomy for treatment of common hepatic duct obstructions associated with duodenal stenosis in two foals. AB - Two female Standardbred foals 2 and 3 months of age were presented with signs of gastroduodenal obstruction that was confirmed with contrast radiography and exploratory surgery. Ventral midline celiotomy was performed, showing stenosis of the duodenum proximal and distal to the hepatopancreatic ampulla. The common hepatic duct, the pancreatic duct, and the sigmoid section of the duodenum proximal to the stenosis were greatly dilated. To bypass the intestinal obstruction, a side-to-side duodenojejunostomy was performed. Obstruction of the common hepatic duct was relieved by side-to-side hepaticojejunostomy. In addition, jejunojejunostomy was performed distal to the other anastomoses. Both foals recovered. On admission, the foals' total bilirubin, aspartate aminotransferase, and gamma glutamyl transferase levels were greatly elevated. During the subsequent 6 to 8 months, they returned to normal. Six months after the first surgery, one foal was readmitted with an acute abdominal crisis. At surgery, there was greater than 360 degrees clockwise rotation of the mesenteric root involving most of the jejunum. At necropsy, the previously created stomas were patent. The liver and bile duct were grossly and histologically normal. The second foal continues to progress normally 12 months after surgery. PMID- 2564708 TI - [Results of a clinical trial of sulfasalazine in rheumatoid arthritis]. AB - The authors tested the therapeutic effectiveness and tolerance of the sulphonamide chemotherapeutic preparation sulfasalazine in 69 patients with active rheumatoid arthritis. A one-year open clinical investigation was completed by 41 patients (59%) where in the course of treatment significant improvement of the majority of clinical and laboratory indicators of the activity of the rheumatic process was recorded. Treatment was terminated prematurely by 28 patients (41%) incl. 16 (23%) where treatment was discontinued because of undesirable side-effects which developed most frequently during the first months of treatment and were not very serious. X-ray evaluation of the joints of the hands after a one-year interval did not reveal deterioration of the X-ray findings in patients who completed the course of treatment. PMID- 2564709 TI - [Possible causes for an increased content of blood peptide hormones in small-cell lung cancer]. AB - The study established the following three possible causes of elevated blood levels of peptide hormones in small-cell lung cancer: endocrine-cell tumor sitting in the lung, presence of tumor endocrine tissues in lung carcinoma of hyperplastic response of normal endocrine cells of the lung and/or other organs to malignancy in the lung. PMID- 2564710 TI - Autoimmune endocrine diseases. PMID- 2564711 TI - Thy-1, CD4 and CD8 in T cell development. PMID- 2564713 TI - Dipeptidyl amino peptidase IV cytochemistry in circulating lymphocytes from HIV-I seropositive subjects. AB - In order to assess whether changes in the relative proportions of cells pertaining to different T-lymphocyte subsets correlate with dipeptidyl amino peptidase IV (DAP IV) activity, the enzyme was cytochemically investigated in circulating lymphocytes from 20 HIV-I-seropositive drug addicts. Although on the average, a significant reduction of lymphocyte DAP IV activity was found in comparison with healthy control subjects, wide variations between individual cases were observed, and no obvious relationship emerged between DAP IV staining on one hand, immunocytological findings and Walter Reed staging on the other. Therefore, in view of the preliminary results reported, the variations of lymphocyte DAP IV activity in the course of HIV I infection seem to deserve further investigation, in order to clarify both the biological significance of this reaction and its possible clinical relevance. PMID- 2564712 TI - Differential growth hormone responses to glucose and growth hormone releasing hormone in lactating dairy cows. AB - Dairy cows in early lactation were reported to secrete growth hormone in response to declining glucose concentrations at day 5 but not day 30 post-partum, whereas GH responses to TRH were reported to be enhanced after day 30 post partum. The present study examined GH response following glucose infusion and the effect of GHRH as well as effects of SRIH on GHRH-stimulated GH release. Declining plasma glucose concentrations after glucose infusion stimulated GH release at day 5 post partum but not in nonpregnant, nonlactating cows or in cows at days 30 and 90 post partum. GHRH stimulated GH release on all days tested, but the response was highest at day 30 post partum when compared to other days. Somatostatin infusion inhibited GHRH effects on GH concentrations only at day 30 post partum and in nonpregnant, nonlactating cows. Thus, a differential response of the GH regulatory system could be demonstrated between days 5 and 30 post partum utilizing different stimuli. Evaluation of plasma glucose and free fatty acid concentrations on days 5, 10, 20, and 30 post partum revealed a progressive decrease in FFA but not glucose as lactation progressed. Decreased plasma FFA concentrations were paralleled by a decrease in basal GH, somatomedin-C and epinephrine. Thus, a decline in FFA may be responsible for the disparity between effects of GHRH and glucose on GH release between days 5 and 30 post partum. PMID- 2564714 TI - Update on beta blockers. AB - Beta adrenergic blockers have been one of the most prescribed category of drugs during the past 15 years. Recently, many new beta blockers have been introduced because they have unique pharmacokinetic and pharmacodynamic properties that benefit patients. The purpose of this article is to review the pharmacological properties and clinical uses of beta blockers. The latter part of this review will discuss the anesthesia implications of this group of drugs. PMID- 2564715 TI - A comparison study of vecuronium bromide and atracurium besylate for rapid sequence induction. AB - Rapid sequence induction is necessary in emergency surgical operations to lessen the chance of aspiration of stomach contents. Succinylcholine usually is the relaxant of choice, because of its rapid onset. However, succinylcholine has side effects which may result in potentially life-threatening conditions. The purpose of this study was to compare two short-acting non-depolarizing muscle relaxants vecuronium and atracurium, using the priming principle, with a depolarizing muscle relaxant, succinylcholine. The comparison may determine if a more suitable method for rapid sequence induction can be identified. Conditions at intubation and at the time to 80-90% neuromuscular blockade were evaluated. Subjects were intubated when the train of four revealed an 80-90% twitch depression. In Group I, the control group using succinylcholine, the mean time to 80-90% neuromuscular block was 74.8 seconds. In Group II subjects, who had received vecuronium, the mean time was 149.4 seconds. Subject in Group III, who received atracurium, had a mean time of 163.7 seconds. There was statistical significance within all three groups (ANOVA, p less than 0.01). Group I subjects showed a significantly faster time to 80-90% neuromuscular block when compared with subjects in Group II and III, but no difference in the time to 80-90% block was revealed between Group II and Group III subjects. Conditions for intubation at 80-90% neuromuscular blockade were the same for all three groups. It was concluded that the administration of vecuronium and atracurium using the priming principle did not allow onset times similar to succinylcholine and that the intubating conditions were similar among all three groups at 80-90% neuromuscular blockade. PMID- 2564716 TI - Vestibular sympathetic nervous system in guinea pig. AB - The vestibular sympathetic fibers were examined in 20 guinea pigs by the immunohistochemical demonstration of tyrosine hydroxylase and dopamine B hydroxylase. The vestibular sympathetics originated in the ipsilateral superior cervical ganglion and entered the internal auditory meatus along the labyrinthine artery. At the Schwann-glial border, some of the sympathetic fibers left the artery and went into the superior and inferior divisions of the vestibular nerve and formed a loose meshwork among the Scarpa's ganglion cells, while other fibers continued to follow the labyrinthine artery. Both groups of fibers entered the cristae ampullares and saccular and utricular maculas after several bifurcations in the cribrose areas and terminated either near the capillaries beneath the sensory epithelia or among the vestibular nerve fibers. These fibers traveled freely in the vestibular labyrinth without being restricted to following blood vessels or vestibular nerve fibers. Some sympathetic fibers made direct contact with the vestibular efferent fibers or the vestibular afferent fibers at the node of Ranvier. Sympathetic fibers were not observed in the sensory epithelia or semicircular canals, and were rarely found in the vicinity of the dark cells. PMID- 2564717 TI - Some aspects of the communicational and computational organization of the brain. AB - Some features of the morphofunctional organization of the CNS have been analysed. Different types of hierarchical organization have been recognized, each of which could deeply affect the circulation (communicational aspect) and elaboration (computational aspect) of information. These two aspects have been discussed on the basis of the existence of two types of electrochemical transmission in the CNS: wiring and volume transmission. By evaluating the CNS operations at different levels of analysis a 'computational hierarchical organization' has been delineated. This concept is very relevant to the understanding of the 'computational power' of the brain (Agnati & Fuxe 1984, Conrad 1985a). In fact, it leads to the distinction between horizontal and vertical elaboration of information. The hypothesis is introduced that in the vertical elaboration of information a central role may be played by the neuronal membrane. In fact, this structure can not only be influenced by the extra- and intracellular signals, but also effectively interconvert the electrical coding into the chemical coding of information. These aspects are discussed in the frame of the possible organization of the membrane into 'domains', each domain being a patch of membrane in which pre-selected molecular movements are possible, resulting in molecular interactions. The movement of a molecule outside its domain is considered energetically unfavourable. The possible formal treatment of this hypothesis is mentioned in Conrad's work (1985b). PMID- 2564718 TI - Basic features of an extracellular recording method to study secretion of a sympathetic co-transmitter, presumably ATP. AB - An extracellular recording method is described which permits in suitable model tissues (e.g. guinea-pig or mouse vas deferens) study of the nerve impulse in sympathetic terminals and the release of transmitter from sites inside or outside the recording electrode. Negative- or positive-going potentials were assumed to reflect the excitatory junction current (EJC) caused by transmitter released inside or outside the electrode, respectively, and hence termed 'EJCi' (i for inside) or 'EJCo' (o for outside). The EJCo were shown to be Ca2+-dependent, blocked by addition of tetrodotoxin or guanethidine, resistant to the alpha 1 adrenoceptor blocking agent prazosin but suppressed by desensitization of P2 purinoceptors by alpha,beta-methylene ATP, and hence, presumably, are caused by release of ATP as a sympathetic co-transmitter. The amplitude of the EJCo was voltage-dependent and increased with the length and frequency of stimulus trains within the range of 1-50 shocks at 0.1-2.5 Hz. In conclusion, combined use of EJCi and EJCo provides a useful tool for physiological and pharmacological analysis of pre- and post-junctional events associated with the secretion of a sympathetic co-transmitter, presumably ATP. PMID- 2564719 TI - Some pharmacological applications of an extracellular recording method to study secretion of a sympathetic co-transmitter, presumably ATP. AB - Extracellular recording in guinea-pig or mouse vas deferens or rat tail artery was used to study the effects of some pharmacological agents on the nerve terminal spike (NTS) and the secretion of a sympathetic co-transmitter (presumably ATP), as reflected in the excitatory junction current (EJC). A negative-going EJCi (i for inside) was assumed to reflect release from sites inside, and a positive-going EJCo (o for outside) release from sites outside the recording electrode. Passage into or out of the electrode seemed to be slow. Tetrodotoxin (TTX) in the outer medium blocked the NTS and ECJo as well as EJCi; TTX in the pipette blocked stimulus-evoked but not spontaneous EJCi. The dihydropyridine Ca2+ channel blocking agent, nifedipine, was without effect, but Cd2+ in the external medium blocked EJCo and also, by an effect apparently 'upstream' of varicosities, inhibited EJCi (i.e. release within the patch) but not the NTS. When present in the outer medium the alpha 2-adrenoceptor agonists, clonidine and xylazine, blocked both EJCo and EJCi, but not the NTS. The effects of clonidine were blocked by yohimbine, which in itself increased the EJCo by about 50%. Neuropeptide Y and met-enkephalin in the outer medium blocked EJCo; the effect of met-enkephalin was blocked by naloxone. The K+ channel blocking agents, tetraethylammonium and 4-aminopyridine, inside or outside the electrode, increased dramatically the size of EJCi or EJCo, respectively. PMID- 2564720 TI - Relationship between renal sympathetic activity and diuretic effects of atrial natriuretic peptide (ANP) in the rat. AB - The effects of various adrenergic agonists and antagonists on the diuretic/natriuretic effects of rANP (103-125) were investigated in conscious and anaesthetized normotensive rats. Pharmacological sympathetic inhibition by reserpine completely inhibited the diuretic/natriuretic effects of ANP. However, surgical renal nerve denervation did not influence the renal response to ANP. Further studies using various pharmacological agents which interfere with adrenergic activity revealed that the diuretic mechanism of action differed between conscious and anaesthetized animals. In the anaesthetized group only, dopamine (D1) blockade reduced ANP-induced diuresis. In the conscious as well as anaesthetized rats, however, pre-synaptic dopamine (D2) stimulation and alpha 2 adrenergic receptor blockade effectively inhibited the renal response to ANP. The results of this study are compatible with the notion that ANP acts indirectly within the kidney via interaction with dopamine-containing neuronal or non neuronal structures in the kidney. PMID- 2564722 TI - Neuroleptic-induced marching-in-place. AB - Marching-in-place (MIP) was observed in 12 out of 133 (9%) chronically hospitalized psychiatric patients and in none of 60 hospital staff controls. The prevalence was similar in both sexes. MIP was associated with tardive dyskinesia or parkinsonism or both but did not occur alone. Counting the number of steps per minute provides a reliable method to quantify MIP and may permit a simple objective method for investigating the neuropharmacology of this phenomenon. Preliminary observations suggest that anticholinergic agents may improve MIP in some patients but worsen it in others. PMID- 2564721 TI - Elevated CSF somatostatin concentrations in demented patients parallel improved psychomotor functions induced by integrity-promoting care. AB - Elderly demented patients from 2 nursing homes participated in this study. An experimental group (n = 17) was subjected to a 3-month program with integrity promoting care resulting in emotional, intellectual and physical activation, whereas a control group (n = 18) had no change from the regular ward care. Dementia rating scales and psychological tests were administered before the start of the study and at the end of the 3-month study period. Lumbar punctures were performed at the same time for assessment of neuropeptide concentrations (somatostatin (SRIF), arginine vasopressin (AVP) and corticotropin-releasing factor) in the cerebrospinal fluid (CSF). In the control group no significant changes were found in the ratings before and after the study. In contrast, improvements in intellectual and motor functioning were observed in the experimental group. Concomitantly, the SRIF concentrations in CSF were significantly elevated in the experimental group and not affected in the control group. The CSF AVP concentrations were reduced in both groups at the end of the study, but considerably more in the control group. It is concluded that environmental factors can improve intellectual and motor functioning in demented patients and also alter CSF biochemical measures of dementia. PMID- 2564723 TI - The effects of terazosin and methyclothiazide on blood pressure and serum lipids. AB - This study compared the antihypertensive efficacy and the effects on serum lipids of terazosin, a new selective alpha 1-adrenergic antagonist, of methyclothiazide (MCTZ), and of the two drugs used as combination therapy. Adult patients with supine diastolic blood pressure ranging from 95 to 120 mm Hg were eligible to enter this double-blind, randomized, parallel-group study. Analyses of the blood pressure data from the 194 evaluable patients revealed that all three treatments produced significant (p less than 0.001) reductions in supine and standing systolic and diastolic blood pressures from baseline values. Moreover, combination therapy resulted in significantly greater mean blood pressure reductions than were observed with either drug used as monotherapy. In the group receiving terazosin monotherapy, the total serum cholesterol level, low-density lipoprotein plus very-low-density lipoprotein cholesterol fraction, and triglyceride level fell significantly (median changes of 3.7%, 5.0%, and 16.3%, respectively, p less than 0.05). However, in the group receiving MCTZ monotherapy, the total serum cholesterol level, low-density lipoprotein plus very low-density lipoprotein cholesterol fraction, and triglyceride level increased significantly (4.7%, 7.1%, and 12.5%, respectively, p less than 0.001). In contrast, no significant changes from baseline values were observed for any lipid variable in the group receiving terazosin/MCTZ combination therapy. We conclude that terazosin is effective antihypertensive therapy that has a potentially beneficial effect on the serum lipid profile when used as monotherapy and that it counteracts the negative impact of MCTZ monotherapy on the serum lipid profile when used concurrently with this thiazide diuretic. PMID- 2564724 TI - Role of the Frank-Starling mechanism in maintaining cardiac output during increasing levels of treadmill exercise in beta-blocked normal men. AB - To determine the effects of beta blockade on hemodynamics during increasing levels of treadmill exercise, 10 healthy volunteers were studied after 1 week of placebo, and then after 1 week of treatment with oral propranolol, 80 mg twice daily, or dilevalol, 400 mg once daily. The study was randomized and double blind, with a crossover sequence. Hemodynamics were measured by CO2 rebreathing at rest and at 25, 50, 75 and 100% of VO2 max. After placebo, cardiac output increased from 5.8 +/- 2.1 (rest), to 19.4 +/- 6.4 liters/min (100% VO2 max), mainly due to an increase in heart rate from 84 +/- 6 to 169 +/- 15 beats/min. Stroke volume increased from 70 +/- 27 (rest), to 137 +/- 65 ml (25% VO2 max), and then leveled off to 116 +/- 41 at 100% VO2 max. After both beta blockers, exercise cardiac output was maintained at 100% VO2 max: 20.1 +/- 9.3 liters/min with propranolol and 19.1 +/- 8.6 with dilevalol. However, a significant reduction versus placebo values was observed for cardiac output at 25% VO2 max, from 13.7 +/- 5.9 during placebo, to 9.4 +/- 2.5 during propranolol, and to 9.6 +/- 2.3 during dilevalol (both p less than 0.01 vs placebo). Maintenance of cardiac output with both beta blockers at higher levels of exercise came from an increased stroke volume (p less than 0.05 vs placebo), while heart rate (in beats/min) was greatly reduced (propranolol 61.6 +/- 9.4 rest, 90.1 +/- 10.7 at 100% VO2 max; dilevalol 70.8 +/- 6.4 rest, 99.2 +/- 11.8 at 100% VO2 max, p less than 0.01 vs placebo for each).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2564725 TI - Esmolol versus verapamil in the acute treatment of atrial fibrillation or atrial flutter. AB - The effects of esmolol, an ultrashort-acting beta blocker, and verapamil were compared in controlling ventricular response in 45 patients with atrial fibrillation or atrial flutter, in a randomized, parallel, open-label study. Patients with either new onset (less than 48 hours, n = 31) or old onset (greater than 48 hours, n = 14) of atrial fibrillation or flutter with rapid ventricular rate were stratified to receive esmolol (n = 21) or verapamil (n = 24). Drug efficacy was measured by ventricular rate reduction and conversion to sinus rhythm. The heart rate declined with esmolol from 139 to 100 beats/min (p less than 0.001) and with verapamil from 142 to 97 beats/min (p less than 0.001). Fifty percent of esmolol-treated patients with new onset of arrhythmias converted to sinus rhythm, whereas only 12% of those who received verapamil converted (p less than 0.03). Mild hypotension was observed in both treatment groups. Esmolol compares favorably with verapamil with respect to both efficacy and safety in acutely decreasing ventricular response during atrial fibrillation or flutter. Moreover, conversion to sinus rhythm is significantly more likely with esmolol. PMID- 2564726 TI - Comparative acute blood pressure reduction from intravenous fenoldopam mesylate versus sodium nitroprusside in severe systemic hypertension. PMID- 2564727 TI - A method for measuring lymphocyte proliferation in mixed lymphocyte cultures using a nuclear proliferation antigen, Ki-67, and flow cytometry. AB - The mixed lymphocyte culture procedure using tritiated thymidine (3H-TdR) incorporation is time consuming and labor intensive, therefore costly. With the use of a fluorescent antibody to a human nuclear proliferation antigen, Ki-67, and flow cytometry, mixed lymphocyte cultures on 20 families of renal and bone marrow transplant patients and normal controls were performed. In this method for measuring lymphocytic proliferation, previously developed by the authors, the entire culture and staining procedures are performed in microculture plates. Finally, the cell suspensions are aspirated with a microsampler to be analyzed by a flow cytometer. Excellent correlation of the percentage of Ki-67-positive cells and the counts per minute (CPM) of 3H-TdR incorporated into the DNA was obtained. This method eliminates the use of radioactive labels, is less time consuming, and yields results two to three days earlier than the radioactive method. In addition, the authors dual-labeled the lymphocyte nuclei with Ki-67 and propidium iodide (Ki-67/PI). This permitted the comparison of the appearance of nuclear antigen with the various phases of the cell cycle. PMID- 2564728 TI - Coronary artery aneurysms in a young adult. AB - Coronary artery aneurysms are an extremely rare cause of death at any age, but particularly so in young adults. A case is described of a young and previously fit man who died suddenly following thrombosis within one of multiple coronary artery aneurysms. He had had a childhood illness with symptoms consistent with an episode of unrecognized infantile polyarteritis nodosa. It is suggested that many cases of aneurysm of the coronary arteries in young adults, where atherosclerosis has been excluded, may in fact be due to previous polyarteritis, rather than of congenital origin as is usually claimed. PMID- 2564729 TI - Molecular genetics of phenylketonuria in Mediterranean countries: a mutation associated with partial phenylalanine hydroxylase deficiency. AB - We report the characterization of a mutation in the phenylalanine hydroxylase (PAH) gene associated with partial residual activity of the enzyme. This point mutation (280glu----lys) was found by sequencing a mutant cDNA clone derived from a needle biopsy of the liver in a child with variant form of phenylketonuria. There is a strict concordance between homozygosity for the mutation and this particular phenotype. The (280glu----lys) mutation is linked to an original and rare RFLP haplotype at the PAH locus found in south Europe and North Africa. So far, this genotype-haplotype association is both inclusive and exclusive. Thirty three PAH-deficient patients were screened for the mutation by using polymerase chain-reaction amplification of their genomic DNA extracted from Guthrie cards. Since a large number of patients can be screened for a particular mutation by using Guthrie cards, the possibility arises of using these samples collected by national newborn screening centers for prospective and retrospective detection of other mutations in the human genome. PMID- 2564730 TI - Linkage disequilibrium study of RFLPs detected at the human muscle nicotinic acetylcholine receptor subunit genes. AB - We screened DNA from unrelated individuals for RFLPs in the muscle nicotinic acetylcholine receptor (AcChoR) genes. These RFLP markers can be used for genetic linkage and association studies to test the hypothesis that receptor structure or regulation is involved in the development of myasthenia gravis (MG). The cDNAs from four subunits (alpha, beta, gamma, and delta) of the murine muscle AcChoR were used as probes to identify RFLPs in the homologous human genes. Digestion of DNA from 15 unrelated individuals with a set of 10 restriction enzymes revealed 11 RFLPs. At least one RFLP was found for each subunit gene. Eight RFLPs were found at the linked gamma and delta gene loci, six with minor allele frequencies greater than 15%, making that linkage group a very informative marker locus (PIC = .72). PIC values were calculated for the RFLPs from allele and haplotype frequency estimates obtained from a population sample of 53 individuals. The delta gene was assigned by in situ hybridization to region q31----q34 of chromosome 2. All pairs of RFLPs were analyzed for linkage disequilibrium. Of the 16 pairs of RFLPs from the same gene or from the linked gamma and delta genes, 13 pairs showed evidence of disequilibrium that was significant, with P less than .05. The implications of these results are discussed. PMID- 2564731 TI - Unequal crossover generates variation in ubiquitin coding unit number at the human UbC polyubiquitin locus. AB - An observed mRNA length polymorphism of the human UbC polyubiquitin gene transcript was shown to correlate exactly with a three-allele HaeIII RFLP. Both polymorphisms apparently result from a variation in the number of ubiquitin coding units per UbC allele. Transcriptionally active alleles containing seven, eight, or nine coding units were observed, at frequencies suggesting that alleles of higher coding-unit number are selectively retained in the population. We propose unequal crossover events, promoted by the highly repetitive structure of the polyubiquitin gene, as a basis for the coding-unit number variation, and we present preliminary evidence for such a crossover, on the basis of analysis of known UbC DNA sequences. PMID- 2564732 TI - Mapping of recombinants near the Huntington disease locus by using G8 (D4S10) and newly isolated markers in the D4S10 region. AB - Genetic linkage between the marker G8 (D4S10) and Huntington disease (HD) was studied in six Dutch pedigrees. The informativeness of the D4S10 locus was increased by isolation of a cosmid, C5.5, with a G8 subclone used as probe. We present a restriction map of 70 kb in the D4S10 region. Two subclones of C5.5, H5.52 and F5.53, detect MspI and SinI RFLPs, respectively. These probes increase the informativeness of D4S10 in the Dutch HD population from 55% to 95%. Seven recombinations were found in 124 informative meioses in which multipoint segregation of D4S10 haplotypes and the HD locus was studied. Two of the recombinations occurred within the D4S10 region. The other five recombinations are highly valuable for the mapping of present and future markers relative to each other and to the HD gene. In addition, several recombinations between markers in meioses from unaffected parents were noted, which will also be useful in ordering new markers. On the basis of our three-point recombination data, the orientation of the D4S10 region relative to HD is HD-H5.52-G8-F5.53, which independently confirms the previously derived polarity for D4S10. PMID- 2564734 TI - Oncogenes and human breast cancer. AB - The role of oncogenes in breast tumorigenesis is unclear. Alterations and/or amplification of several oncogene sequences have been observed in primary human breast tumors, in breast tumor cell lines, and in mammary tumors in model systems. In principle, such alterations could be sites of primary lesions for human breast cancer, causes of tumor progression or metastasis, or simply secondary lesions of highly aberrant tumor genomes. The present study tested genetic linkage of breast cancer susceptibility to nine oncogenes in 12 extended families including 87 affected individuals. Lod scores for close linkage of each candidate sequence to breast cancer were -19.6 for HRAS, -12.3 for KRAS2, -1.0 for NRAS, -6.0 for MYC, -6.1 for MYB, -8.2 for ERBA2, -7.9 for INT2, and -5.1 for RAF1. Regions of chromosome 11p associated with tumor homozygosity and the region of 3p carrying the gene for Von Hippel-Lindau disease could also be excluded from linkage to human breast cancer. The 5-kb allele of the MOS oncogene, previously proposed to be associated with breast cancer, was absent in these families, suggesting that polymorphism at this locus is not associated with inherited susceptibility. These results strongly suggest that oncogenes are not the sites of primary alterations leading to breast cancer. On the other hand, alterations in one or more of these sequences may be associated with tumor progression. PMID- 2564733 TI - Autosomal dominant retinitis pigmentosa: exclusion of the gene from the short arm of chromosome 1 including the region surrounding the rhesus locus. AB - Members of a large Irish pedigree exhibiting early-onset autosomal dominant retinitis pigmentosa (ADRP) were typed for the rhesus blood group and nine DNA markers on chromosome 1. Close linkage between the ADRP locus and any of the marker loci was excluded using two-point analysis. With use of the sex-averaged maps of Dracopoli et al. and Donis-Keller et al. and a strategy of rolling multipoint analyses, support was gained for the exclusion of ADRP from a 224-cM region of the chromosome, including almost the entire short arm. The disease locus was significantly excluded from within at least 50 cM of the rhesus locus and, as a loose linkage between these two genes has been suggested by other studies, this result may support the possibility of genetic heterogeneity within the autosomal dominant subgroup of retinitis pigmentosa. PMID- 2564735 TI - Isolated systolic hypertension in the elderly: results of a statewide survey of clinical practice in New Jersey. AB - PURPOSE AND SUBJECTS AND METHODS: The advisability of treating isolated systolic hypertension (ISH) in the elderly is a matter of debate. We therefore surveyed current clinical practices for ISH among members of the New Jersey Academy of Family Physicians and the American College of Physicians-New Jersey Chapter (n = 1,514). To our knowledge, no such survey focusing on ISH had previously been reported. Two rounds of questionnaires were mailed three weeks apart in April and May of 1985, followed by a phone survey of non-respondents in June and July. A third mailing was performed where appropriate. RESULTS: The response rate was 87 percent. For ISH in patients aged 60 and older, 89 percent of the 1,318 respondents reported using drug therapy. A diuretic was chosen as the first-line drug by 67.5 percent of respondents. The most common second-line drug choices were a beta blocker (28 percent), a central alpha agonist (19 percent), and a diuretic (18.5 percent). The patient's age entered strongly into decisions on drug use. Although 55 percent of respondents would use drug therapy at a systolic blood pressure of 160 mm Hg or less for persons aged 60 to 69, only 35 percent would do so for persons aged 70 to 79, and 26 percent for persons aged 80 and older. For all patient age groups, older physicians were more likely to consider drug therapy than were younger physicians. Among specialty groups, general practitioners were the most likely to consider drug therapy and cardiologists the least likely to do so for all patient age groups. Family physicians and general internists appeared to have very similar practice patterns regarding definition and treatment of ISH. CONCLUSION: The overall message of the survey results is that the majority of physicians questioned use drug therapy, usually a diuretic or a beta blocker, for some patients with ISH. However, no widely accepted standard practice on the definition and management of ISH in persons aged 60 and older was observed. PMID- 2564736 TI - Growth hormone deficiency, wormian bones, dextrocardia, brachycamptodactyly, and other midline defects. AB - We report on a 17-month-old boy with Wormian bones, short stature, growth hormone deficiency, developmental delay, brachycamptodactyly, dextrocardia, cryptorchidism, midshaft hypospadias, hypoplastic left kidney, and imperforate anus. This unique combination of abnormalities has not been reported previously. PMID- 2564737 TI - McKusick-Kaufman syndrome: report of an instructive family. AB - We report on two sisters and their brother, all of whom are affected by McKusick Kaufman syndrome. This sibship confirms autosomal recessive inheritance and the occurrence of the syndrome in Melanesians. It also demonstrates multisystem involvement in both sexes and illustrates difficulties in the diagnosis and management of hydrocolpos. PMID- 2564738 TI - Prenatal exclusion testing for Huntington disease using the polymerase chain reaction. AB - Prenatal exclusion of Huntington disease (HD) may be carried out by analysis of cosegregating DNA markers on a first-trimester chorionic villus sample. The conventional Southern blot method is time-consuming and requires microgram quantities of DNA and milligram quantities of villus tissue. The use of the polymerase chain reaction (PCR) to amplify genomic DNA by a factor of 10(7) or more makes it possible to do analyses on very small samples in a few hours and without recourse to Southern blotting or hybridization with radioactive probes. We report on a fetus at risk of HD; prenatal testing was carried out by using the PCR to amplify a polymorphic DNA sequence adjacent to the HD locus. The risk of the fetus inheriting the HD gene could not be excluded and the pregnancy was terminated. This represents an example of gene tracking by using amplification of a restriction fragment length polymorphism at some distance from the relevant mutation. PMID- 2564739 TI - Angelman and Prader-Willi syndromes share a common chromosome 15 deletion but differ in parental origin of the deletion. AB - Many Prader-Willi syndrome (PWS) and Angelman syndrome (AS) patients have a cytogenetic deletion of 15q11q13. While AS and PWS share a similar cytogenetic anomaly, they have very different clinical phenotypes. DNAs from 4 AS patients were examined using 5 chromosome 15q11q13-specific cloned DNA segments. With the present level of resolution, the molecular deletions between AS and those previously reported for PWS did not appear to differ. However, in contrast to the paternal inheritance of the deleted chromosome 15 observed in the majority of PWS patients, maternal inheritance of the deleted chromosome 15 was demonstrated in the AS patients by restriction fragment length polymorphisms (RFLPs). PMID- 2564740 TI - Polyarteritis nodosa in pregnancy: a case report and brief literature review. AB - Patients who develop polyarteritis nodosa in pregnancy generally have a grim prognosis. A young nulliparous woman developed the disease and attained remission before becoming pregnant. Although she developed a recurrence, both mother and infant did well. It appears that if a patient becomes pregnant while in remission the chance of a successful pregnancy outcome is reasonable. PMID- 2564741 TI - Bolus tocolysis: treatment of preterm labor with pulsatile administration of a beta-adrenergic agonist. AB - The treatment of premature labor with beta-adrenergic substances is complicated by side effects. Although most human control mechanisms are pulsatile, therapy is usually administered continuously. We designed a microprocessor-controlled pump to allow pulsatile tocolytic infusion, hoping to reduce the total dose and thus the side effects. In 33 patients pulsatile bolus tocolysis was compared with continuous tocolysis in a control group of 38 patients. Bolus tocolysis required considerably less beta-sympathomimetic agent for comparable therapeutic success (median dosage 3.0 versus 15.9 mg, p less than 0.001). Duration of therapy under bolus tocolysis was also significantly shorter (p less than 0.05). Birth weight was higher after bolus tocolysis (median 3070 versus 2580 gm, p = 0.05). Additional indicators favored bolus tocolysis but were not statistically significant: a longer gestational period, fewer infants weighing less than 2500 gm, and a lower incidence of respiratory distress syndrome. Pulmonary edema occurred in one patient during continuous tocolysis. PMID- 2564742 TI - Zygosity determination of multiple pregnancy by deoxyribonucleic acid fingerprints. AB - We used a new method of deoxyribonucleic acid analysis to determine zygosity in multiple pregnancies. This method uses a minisatellite core probe, requires only a small amount of deoxyribonucleic acid, and detects the restriction fragment length polymorphisms that are a result of allelic differences in the number of tandem repeats that contain the core sequence. Southern blot hybridization showed an individual-specific deoxyribonucleic acid fingerprint and each polymorphic band in the sibling could be identified within one (but not both) of the parents. Identical deoxyribonucleic acid fingerprints among the siblings of multiple pregnancy indicate they must be monozygotic. This method is sufficiently reliable and rapid so the determination of zygosity in multiple pregnancy can be made the same day the fetal deoxyribonucleic acid is made available. PMID- 2564744 TI - [Infection of bones and joints]. PMID- 2564743 TI - Addiction to neuroleptics? PMID- 2564745 TI - Impairment of the antagonism of vecuronium-induced paralysis and intra-operative disopyramide administration. AB - A 63-year-old male was admitted to hospital for a cholecystectomy, vagotomy and gastro-enterostomy. Muscle paralysis was induced with 70 micrograms/kg vecuronium, followed by increments of 20 micrograms/kg when the initial twitch height returned to 25% of control. The patient received 3 doses of 10 mg disopyramide intravenously, on account of supraventricular ectopic beats, followed by an infusion of 25 mg/hour. Paralysis was reversed using 0.75 mg atropine and 2.5 mg neostigmine once the twitch height had returned spontaneously to 25% of its initial value. Fifteen minutes later, twitch height had returned to control value and the train-of-four was above 85%, but the responses to tetanic stimulation at 100 Hz and 50 Hz remained severely depressed (10% and 45%, respectively). The patient's trachea was extubated after 20 minutes, but residual fade was still observed. This impairment of neuromuscular transmission, detected only with high frequency stimulation, was present with a measured concomitant plasma level of disopyramide of 5.1 micrograms/ml. PMID- 2564747 TI - The use of perioperative opiates for laparoscopy. PMID- 2564746 TI - The use of flumazenil (Anexate, Ro 15-1788) in the management of drug overdose. AB - This is a report of a large selfadministered overdose of temazepam and meprobamate. The administration of flumazenil (Ro 15-1788) led to the partial antagonism of the depressant action of the drugs which was sufficient to avoid the need for invasive respiratory and cardiovascular support. PMID- 2564748 TI - Anxiety and informed consent. PMID- 2564749 TI - Doses of muscle relaxants. PMID- 2564750 TI - Interaction between nitrous oxide and diazepam in the mouse staircase test. AB - The interaction between nitrous oxide and diazepam was assessed in the mouse staircase test. In this paradigm, the numbers of rears (NR) (simultaneous standing on hindlegs and sniffing the air) and steps ascended (NSA) reflect anxiety and locomotor activity, respectively. Antianxiety drugs characteristically reduce NR but not NSA, while sedative drugs reduce both NR and NSA in generally parallel fashion. In this study, increasing concentrations of nitrous oxide alone produced progressively greater and statistically significant increases in NSA. Nitrous oxide alone had no significant effect on NR until, at a concentration of 75%, it decreased NR. Diazepam alone decreased NR equally and significantly at all doses. With certain combinations of concentration and dose, nitrous oxide enhanced the effect of diazepam on NR without altering NSA. These findings suggest that nitrous oxide can increase the antianxiety effect of diazepam without increasing its sedative effect in the mouse staircase paradigm. PMID- 2564751 TI - Prolonged vecuronium-induced neuromuscular blockade in children. PMID- 2564752 TI - Effect of oral and intramuscular famotidine on pH and volume of gastric contents. PMID- 2564753 TI - Benzodiazepine antagonism does not provoke a stress response. AB - Acute anxiety reactions have been reported following antagonism of benzodiazepine induced sedation. In this study, the level of sedation and anxiety was assessed in 30 patients randomly assigned to receive either saline or flumazenil (a benzodiazepine antagonist) after midazolam sedation according to a double-blind protocol. Carefully titrated doses of flumazenil, 0.8 +/- 0.2 mg (mean +/- SD), effectively reversed residual midazolam-induced sedation without producing significant changes in the patients' level of anxiety. In addition, plasma epinephrine, norepinephrine, vasopressin, and beta-endorphin concentrations were measured in a subset of patients (n = 5) from each group. The levels of these stress hormones did not acutely change following flumazenil (or saline). These results indicate that flumazenil, 0.6-1.0 mg iv, can antagonize midazolam sedation without producing acute anxiety or evidence of a stress response. PMID- 2564754 TI - A postoperative pain management service. PMID- 2564755 TI - The function of synaptic transmitters in the retina. PMID- 2564756 TI - Involvement of hormonal and neuromodulatory systems in the regulation of memory storage. PMID- 2564757 TI - Acute regulation of tyrosine hydroxylase by nerve activity and by neurotransmitters via phosphorylation. PMID- 2564758 TI - [Hemorrhagic gastroduodenal ulcer: contribution of new medical treatments]. AB - The mortality due to upper gastrointestinal haemorrhage from ulcers has remained unchanged for several (estimated to be 6%), despite progress in surgery and intensive care, new drug treatments and, more recently, the development of endoscopic treatments. Drug treatments, although frequently used in practice always have a controversial haemostatic efficacy. Antacids have a haemostatic efficacy, but only at doses responsible for side effects. Antisecretory drugs appear to be systematically prescribed, if only to commence treatment of the peptic ulcer disease. However, their haemostatic efficacy has not been established except in the sub-group of gastric ulcers, especially in elderly subjects. According to a recent study, prostaglandins are as active as antacids. Since 1975, endoscopy is the support for haemostatic treatments, most of which are still under evaluation. Therapeutic injections, a simple and inexpensive technique, certainly represents one of the most promising methods. Different authors have proposed different solutions for injection (absolute alcohol, aetoxysclerol, adrenaline, thrombin...) with comparable haemostatic efficacy. Laser photocoagulation (Argon and Nd-YAG) is an expensive and impractical technique and a number of randomised studies indicate that the initial enthusiasm for this technique is not justified. Monopolar electrocoagulation does not constitute a real progress in contrast with bipolar electrocoagulation which significantly decreases the frequency of recurrent haemorrhages. Lastly, the efficacy of the heater probe remains to be evaluated in the light of promising studies, particularly those of Johnson in 1985. At the present time, these endoscopic haemostatic treatment should be reserved to high surgical risk patients (elderly or debilitated subjects).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2564759 TI - Chemical ecology and behavioral aspects of mosquito oviposition. PMID- 2564760 TI - Asthma, wheezing, and school absence in primary schools. AB - The prevalence of wheezing, 'asthma', treatment for 'asthma', and school absence as a result of wheezing in Nottingham was calculated from a questionnaire survey of parents of 4750 children in a random sample of primary schools. A response was achieved for 3805 (80%) children of whom 438 (11.5%) had had episodes of wheezing in the last year and 224 (5.9%) had been diagnosed as having asthma. Asthma treatment had been prescribed for 251 (6.6%) of all children, two thirds of all the children receiving drugs. Two hundred and sixty five (7%) children had lost time from school because of wheezing (median loss of seven days). Of the 64 children losing more than 10 days, 45 (70%) were not taking any drugs, or taking only beta agonists. The prevalence of wheezing found by this survey was comparable with that in similarly designed surveys, though the proportion of children diagnosed as having asthma was higher. Though doctors may now diagnose asthma more readily wheezing still remains an important cause of school absence and still seems to be undertreated. PMID- 2564761 TI - Schizophrenia in the elderly: what nurses need to know. AB - People age 65 and older with schizophrenia comprise a small but growing number of the mentally ill that nurses, especially those who work in nursing homes, care for. Elderly schizophrenics are a heterogeneous group. They are subject to all of the chronic and acute illnesses common to elderly persons. In addition, their medical problems are frequently overlooked and are difficult to treat. As many as 50% of them have tardive dyskinesia, which has potentially fatal consequences, especially in the elderly. PMID- 2564762 TI - Effects of alpha 2-adrenoceptors blockade on the inhibition of the nociceptive jaw-opening reflex by the lateral reticular nucleus in rabbits. PMID- 2564763 TI - Pharmacokinetic determinants of dynamic differences among three benzodiazepine hypnotics. Flurazepam, temazepam, and triazolam. AB - Healthy adult volunteers (n = 52) received single oral doses of flurazepam hydrochloride (15 mg), temazepam (15 mg), triazolam (0.25 mg), or placebo in a parallel, double-blind study. Sedative effects were greatest with triazolam, followed next by temazepam; peak effects closely coincided with peak plasma concentrations. Differential recovery from sedation corresponded in part to differences in mean elimination halflife, although sedative effects returned to baseline before plasma drug concentrations became undetectable. Sedation following flurazepam administration was less intense than with triazolam and temazepam. When tested at three hours after dosing, none of the active treatments impaired learning of a 16-item word list. However, at 24 hours, triazolam recipients could not recall a significant fraction of what was learned. Thus, dynamic differences among three benzodiazepine hypnotics may be partly explained by kinetic differences, as well as, we should caution, by possible "clinical inequivalence" in dosage. PMID- 2564764 TI - Atypical neuroleptics, dose-response relationships, and treatment-resistant psychosis. PMID- 2564765 TI - Parkinsonism following neuroleptic withdrawal. PMID- 2564766 TI - Ethics of drug discontinuation studies in schizophrenia. PMID- 2564768 TI - Pisa syndrome in a Chinese patient. AB - A Chinese female with 10 years' history of a special form of tardive dystonia (Pisa syndrome) is described. Mild improvement has occurred after 5 months' treatment with tetrabenazine despite the long duration of dystonia and the presence of secondary structural changes. PMID- 2564767 TI - Inhibition of the stimulated canine exocrine pancreas by amino acids and fat. AB - To investigate the possible exocrine pancreatic inhibitory actions of amino acids and fat, pancreatic fistula outputs and plasma concentrations of glucagon, somatostatin, and pancreatic polypeptide were measured for response to intravenous (IV) and intraduodenal nutrient administration in six dogs submaximally stimulated with cholecystokinin. Intravenous amino acids caused abrupt and significant 45% to 73% reductions in stimulated pancreatic protein, bicarbonate, and volume outputs. There were no significant associated changes in plasma hormone concentrations and no similar immediate pancreatic inhibition with IV mannitol, thus suggesting a possible direct inhibitory effect of amino acids. Intraduodenal amino acids and IV fat evoked no significant pancreatic output suppression. Intraduodenal fat rapidly caused a significant 40% to 62% reductions in stimulated outputs that were associated with an 81% rise in plasma pancreatic polypeptide concentration, suggesting a gut-mediated inhibition. We conclude that IV amino acids and intraduodenal fat both inhibited stimulated pancreatic secretion but probably by different mechanisms. PMID- 2564769 TI - Tardive dyskinesia clinic. PMID- 2564770 TI - College congress. PMID- 2564771 TI - Different forms of post-training memory processing. AB - Memories are not acquired in their definitive form, but can be considerably modified in the period that follows after acquisition, both quali- and quantitatively. This may happen either by a process of consolidation or strengthening of each memory trace or by the incorporation of further information to each experience. This further information may be provided by the action of drugs, including that of endogenous substances released by each training experience, and by the addition of information provided by other tasks or events. Evidence in favor of the existence of these various post-training processes, and of their importance for memory formation, is discussed. All of these processes are time dependent, all may depend on stimulus aftereffects, each is differently affected by drugs given in the post-training period, and all may interact with each other. PMID- 2564772 TI - Applied tension, applied relaxation, and the combination in the treatment of blood phobia. AB - Thirty patients with phobia for blood, wounds and injuries were treated individually with applied tension, applied relaxation, or the combination of these two methods for 5, 9 and 10 sessions, respectively. They were assessed on self-report, behavioral and physiological measures before and after treatment, and at a 6-month follow-up. All groups improved significantly on 11/12 measures, and the improvements were maintained at follow-up. Applying stringent criteria, 73% of the patients were clinically improved at the end of treatment and 77% were so at follow-up. Despite a failure to find between-group differences, on many measures there was a trend favoring applied tension. Since this method is as effective as the other treatments in only half the time, applied tension should clinically be the treatment of choice for blood phobia. PMID- 2564773 TI - Relationships among measures of communication, marital satisfaction and exposure during couples treatment of agoraphobia. AB - The goals of this study were to examine the relationships among measures of couples interactions (satisfaction and communication), and response to treatment within a group of agoraphobic clients and their partners. The treatment included the clients' partners in structured, graduated in vivo exposure practices. Twenty two clients and their partners completed pre-, mid- and post-treatment assessments. Treatment responders rated themselves and their partners as more communicative regarding the client's fears, at pre- and mid-assessments in comparison to nonresponders; measures of communication related inversely to levels of anxiety reported during exposures, but marital satisfaction did not relate to any measure of exposure. In addition, level of anxiety during exposure differentiated responders from nonresponders; responders experienced a significant reduction in their levels of anxiety. Finally, ratings of frequency of communication at mid-assessment were highly predictive of treatment outcome at post-assessment. The implications from these findings for communication skills training for a subset of couples who enter couples treatment for agoraphobia are discussed. PMID- 2564774 TI - Tb(III)-ion-binding-induced conformational changes in platelet factor XIII. AB - Ca(II) ions are crucial during proteolytic conversion of Factor XIII zymogen into the active enzyme Factor XIIIa. Factor XIII proteolyzed by thrombin or trypsin in the presence of 5 mM-EDTA resulted in rapid inactivation of transglutaminase activity. Factor XIIIa formed by thrombin or trypsin in the presence of 40 microM Tb(III) ions, however, was indistinguishable from Factor XIIIa formed in the presence of 2-5 mM-Ca(II) ions with respect to molecular mass and transglutaminase activity. Thrombin treatment of Factor XIII in the presence of 1 5 microM-Tb(III) ions resulted in three fragments (76 kDa, 51 kDa and 19 kDa) with simultaneous loss of transglutaminase activity. Tb(III) ions at concentrations greater than 40 microM made platelet Factor XIII resistant to proteolysis by either thrombin or trypsin. Other lanthanide(III) ions [Ln(III) ions] tested [Ce(III), La(III) and Gd(III) ions] functioned similarly to Tb(III) ions during proteolytic activation of Factor XIII. Ln(III) ions (10-100 microM) were unable to replace the Ca(II) ions required for transglutaminase activity of Factor XIIIa. Tb(III) ions also inhibited in a non-competitive manner the transglutaminase activity of Factor XIIIa (Ki 71 microM) even when measured in the presence of 200-fold molar excess of Ca(II) ions. Factor XIII selectively bound to a Tb(III)-chelate affinity column, and could not be eluted by 100 mM CaCl2. Binding of Tb(III) ions to Factor XIII was demonstrated by fluorescence emission due to Forster energy transfer. A 10(4)-fold molar excess of CaCl2, but not NaCl, partially quenched Tb(III) fluorescence. Low concentrations (5-20 microM) of Tb(III) ions also inhibited the binding of Factor XIII to des-A fibrinogen by about 43%, whereas higher concentrations (40-100 microM) promoted binding. Conformational changes in Factor XIII consequent to the binding of Tb(III) ions could be responsible for the observed effects on protein structure and function. PMID- 2564775 TI - 13C-n.m.r. study of citrate metabolism in rabbit renal proximal-tubule cells. AB - The metabolism of a 13C-labelled substrate, [3-13C]citrate, was monitored in rabbit renal proximal-tubule cells by 13C n.m.r. The relative enrichments of label in glutamate, glutamine and alpha beta-glucose allowed a calculation of the rate of the glutamate dehydrogenase flux relative to the flux via phosphoenolpyruvate carboxykinase. A ratio of 1.2 +/- 0.05 was found. Under steady-state conditions of active gluconeogenesis, the ratio of flux through pyruvate kinase to the gluconeogenic rate was 0.97 +/- 0.03. PMID- 2564776 TI - Evidence that the production of acetate in rat hepatocytes is a predominantly cytoplasmic process. AB - By using [1-14C]butyrate, the fluxes of butyrate to acetate and fatty acids were measured in rat hepatocytes. Both fluxes were inhibited to a similar extent by ( )-hydroxycitrate, with no significant effect on butyrate uptake. These results indicate that acetate formation takes place in the cytoplasm, presumably via ATP stimulated acetyl-CoA hydrolase. Since acetate formation occurred despite a net uptake of acetate, the results are also consistent with the operation of a substrate cycle between acetate and acetyl-CoA, recently proposed by other workers, and suggest that this cycle is cytoplasmic. PMID- 2564777 TI - Bovine milk-fat-globule membrane contains an enzymically inactive form of acetyl CoA carboxylase. AB - Enzymically inactive acetyl-CoA carboxylase [acetyl-CoA:carbon-dioxide ligase (ADP-forming), EC 6.4.1.2] was found as a component of bovine milk-fat-globule membrane (MFGM). Acetyl-CoA carboxylase was present in MFGM at a higher concentration than in cytosolic or mitochondrial fractions of bovine mammary tissue, which makes it unlikely that its presence was due to simple contamination by these subcellular constituents. PMID- 2564778 TI - Somatostatin and lanthanum discriminate two Ca2+ mobilizing mechanisms regulated by bombesin receptors in peptic cells. AB - Bombesin (BB)-stimulated pepsinogen secretion from frog esophageal peptic acini was inhibited 40% by somatostatin (SS) (IC50 = 1 nM), and by 30-50% by low concentrations (10(-7)-10(-4)M) of lanthanum chloride. SS inhibited basal secretion as well as both early (0-2 min) and late (2-30 min) BB-stimulated secretory phases. By contrast, LaCl3 selectively inhibited the late secretory phase and was without effect on basal secretion. SS (100 nM) and LaCl3 (30 microM) attenuated BB-stimulated 45Ca2+ uptake, and the combination resulted in additive inhibition. High K+ media decreased basal secretion, abolished SS but not LaCl3 inhibition of BB-stimulated secretion, and blocked SS inhibition of BB mediated 45Ca2+ uptake. These findings suggest the existence on peptic cells of distinct La3+-sensitive, and somatostatin-sensitive, K+ dependent, Ca2+ mobilizing mechanisms which contribute to BB receptor-mediated secretion. PMID- 2564779 TI - Autoradiographic analyses of agonist binding to muscarinic receptor subtypes. AB - The binding of four muscarinic receptor agonists to regions of rat brain was examined through quantitative autoradiographic techniques. Oxotremorine, arecoline, pilocarpine and bethanechol were chosen based on their different potencies and efficacies in muscarinic second messenger systems. Overall, the order of potency for inhibition of [3H]-l-quinuclidinyl benzilate ([3H]-l-QNB) binding to rat brain slices was oxotremorine greater than pilocarpine = arecoline much greater than bethanechol. Regional assays of agonist potency indicated that all agonists were more selective for brainstem and thalamic regions than for hippocampal and cortical regions. The high selectivity of agonists for areas such as the paraventricular thalamus and the superior colliculus, which also display low affinity for pirenzepine, suggests that muscarinic agonists bind with higher affinity to M2 receptors. Of the four agonists examined, pilocarpine displayed the lowest selectivity for M2 receptors in that IC50 values for pilocarpine were only 3-fold higher in the hippocampal and striatal regions (e.g. CA3: 40.6 +/- 9.4 microM) than in thalamic and brainstem regions (e.g. paraventricular thalamus: 14.9 +/- 6.2 microM). Oxotremorine was 8-fold more potent in the brainstem and thalamus, while arecoline and bethanechol were, respectively, 19- and 100-fold more selective for brainstem and thalamic receptors. Scatchard analyses revealed heterogeneous binding profiles for some agonists within single brain regions, suggesting that multiple agonist sites exist even within regions of predominantly M1 or M2 receptors. For example, arecoline displayed curved Scatchard plots within the external layers of the cerebral cortex, layer CA1 of the hippocampus (predominantly M1 subtype), and the paraventricular thalamus (predominantly M2 subtype). The ability of agonists to recognize multiple sites within a single region may reflect the ability to recognize receptors coupled or uncoupled to second messenger systems through G-proteins. PMID- 2564780 TI - Elevated levels of serum hyaluronate and correlation with disease activity in experimental models of arthritis. AB - The serum hyaluronate (HA) concentration was measured in groups of rats immunized for adjuvant or type II collagen arthritis. Serum HA increased as the arthritic lesions developed, correlating with the severity of the disease. This increase in HA was not related to metabolic impairment, because rats with adjuvant arthritis metabolized intravenously administered tritiated HA at a rate similar to that of normal rats. Serum HA levels may be useful as an indicator of synovitis in experimental and possibly in clinical arthritis. Further, this model could serve as an experimental approach for studies of HA metabolism in chronic joint inflammation. PMID- 2564781 TI - Requirements for flare reactions of joint inflammation induced in mice by cloned MT4+, Lyt-2- T cells. AB - Joint inflammation was induced in C57B1/6 mice by injection of cloned MT4+, Lyt-2 T cells specific for the antigen methylated bovine serum albumin (mBSA), together with mBSA. In this model, after waning of the inflammation, flare reactions can be induced by a rechallenge with the specific antigen. Herein we show that such flare reactions can still be induced several weeks after waning of the joint inflammation, as was demonstrated both in normal C57B1/6 mice and in athymic C57B1 nude mice. The results in the latter group indicate that T cells of the recipient mice are not necessary for the elicitation of flare reactions. On histologic examination, the inflammatory infiltrates in the knee joints of the nude mice appeared to be mainly granulocytic. The cloned T cells persisted and remained functionally reactive in the knee joint for at least 2 weeks in the absence of the antigen, and thus, in the absence of inflammation. In view of the similarities between induced joint inflammation in mice and rheumatoid arthritis in humans, these data may be relevant to our understanding of the processes involved in the latter disease. PMID- 2564782 TI - Recovery of neuromuscular function after infusion or intermittent bolus doses of atracurium or vecuronium. AB - Neuromuscular block and postoperative recovery of grip strength and peak expiratory flow (PEF) were compared in patients receiving atracurium or vecuronium administered by continuous infusion or intermittent bolus doses. The same total dose of atracurium (0.92-0.98 mg kg-1) or vecuronium (0.16-0.18 mg kg 1) was given by both methods. A similar degree of neuromuscular block was attained in all groups. A control group receiving no neuromuscular blocking drugs was also studied. Grip strength and PEF were reduced significantly in all groups (less than 80% of preoperative value) 15 min after operation. This was most marked following infusion of vecuronium (less than 50%). Grip strength recovered in all groups in 30-60 min. PEF was still significantly less than control value at 90 min in all groups receiving neuromuscular blocking drugs. PMID- 2564783 TI - Train-of-four fade during onset and recovery of neuromuscular block: a study in non-anaesthetized subjects. AB - The actions of alcuronium, vecuronium and tubocurarine have been studied in the isolated forearms of six healthy, non-anaesthetized volunteers. The responses of adductor pollicis were measured during onset and recovery of neuromuscular block for each agent. There was a drug-related disparity between mechanomyogram (MMG) and electromyogram (EMG) measurement of the first response of the train-of-four (T1) and of the ratio of the fourth (T4) to the first response (TOF ratio). There were significantly higher T1 values for the EMG than for MMG during alcuronium blockade (P = 0.03). For tubocurarine, however, the relationship was reversed. The relationship between T1 and TOF ratio during onset and recovery of neuromuscular block was a hysteresis. The TOF ratio at 50% T1 was significantly higher during onset than during recovery for all three drugs, measured by MMG or EMG (P less than 0.005). Analysis of variance of the differential fade loops failed to show a drug-related effect. We conclude that care should be taken in assuming interchangeability between MMG and EMG measurement of T1. Relationships between T1 and TOF ratio derived during recovery do not necessarily apply during onset and may lead to error in estimating the degree of muscle relaxation. PMID- 2564784 TI - Effects of pipecuronium and pancuronium on the isolated rabbit heart. AB - We have studied the effects of pipecuronium and pancuronium on myocardial contractility and heart rate in two different animal preparations. Pipecuronium and pancuronium produced no change in isometric contraction of rabbit atria. The chronotropic effects of pipecuronium, pancuronium and vecuronium were investigated using acetylcholine as an agonist in isolated perfused rabbit heart. Pancuronium but not pipecuronium or vecuronium, produced a significant degree of antagonism to the bradycardia produced by acetylcholine. PMID- 2564785 TI - Influence of suxamethonium on the action of subsequently administered vecuronium or pancuronium. AB - The effects of suxamethonium were studied on the onset time and duration of action of vecuronium or pancuronium in 45 adult patients anaesthetized with halothane and nitrous oxide. After an intubating dose of suxamethonium, the force of the evoked twitch returned to a value greater than control. The onset of the reduction in force produced by subsequent administration of vecuronium or pancuronium was faster and recovery slower. This potentiating effect of suxamethonium persisted for at least 2 h. PMID- 2564786 TI - Schizophrenia, tardive dyskinesia, and brain GABA. AB - We measured the contents of gamma-aminobutyric acid (GABA) and of other amino compounds in five regions of autopsied brain from 18 patients with schizophrenia and from a large group of adult control subjects dying without any neurological or psychiatric disorder. In addition, concentrations of GABA were measured in the cerebrospinal fluid (CSF) of living schizophrenic patients and control subjects. No deficiency of GABA was found in the frontal cortex, caudate nucleus, putamen, nucleus accumbens, or medial dorsal thalamus of patients dying with schizophrenia, nor were GABA concentrations low in the CSF of living schizophrenic patients. These results do not confirm our earlier report of low levels of GABA in the nucleus accumbens and thalamus of some schizophrenic patients. We do not find neurochemical evidence favoring an involvement of GABAergic neuronal hypofunction in the etiology either of schizophrenia or of neuroleptic-induced tardive dyskinesia. PMID- 2564787 TI - Fluphenazine treatment reduces CSF somatostatin in patients with schizophrenia: correlations with CSF HVA. AB - CSF somatostatin and homovanillic acid (HVA) were measured in 14 schizophrenic patients while they were drug-free and during chronic fluphenazine treatment. CSF somatostatin was significantly reduced and CSF HVA was significantly elevated (p less than 0.002) during fluphenazine treatment. There was a trend toward correlation between CSF somatostatin and CSF HVA in the 14 schizophrenic patients when drug-free (r = 0.49, p less than 0.07) and fluphenazine-treated (r = 0.47, p less than 0.08). When examined in a larger group (n = 46) of drug-free schizophrenics, this relationship was highly significant (r = 0.59, p less than 0.001). These clinical data are consistent with preclinical evidence indicating a functional interaction between CNS somatostatin and dopamine systems. PMID- 2564788 TI - Comparison of prolactin responses to D-1 and D-2 antagonists in rats: Ro 22-1319 is a potent D-2 antagonist. PMID- 2564789 TI - Molecular organization of the cytokine gene cluster, involving the human IL-3, IL 4, IL-5, and GM-CSF genes, on human chromosome 5. AB - The human genes for the hematopoietic growth factors interleukin-3 (IL-3), IL-5, and granulocyte-macrophage colony-stimulating factor (GM-CSF) have been mapped to 5q23-31. We present in situ hybridization evidence that the human IL-4 gene is located at 5q23.3-31.2, suggesting that the four cytokine genes may be closely linked. We used pulsed-field gel electrophoresis to prepare subchromosomal restriction maps surrounding these genes to define this possible linkage more precisely. The IL-4 and IL-5 genes are tightly linked, being 90 to 240 kilobases (kb) apart, as has been shown for the IL-3 and GM-CSF genes, which are only 9 kb apart. Possible overlap of the map containing the IL-4 and IL-5 genes with restriction sites 5' to the IL-3 gene suggests that the four cytokine genes may be localized within 500 kb of each other. The endothelial cell growth factor gene (ECGF), which has also been localized to the 5q31 region, did not appear to be close to the cytokine genes. Linkage of the IL-3, IL-4, IL-5, and GM-CSF genes has important implications in the evolutionary origin and regulation of expression of these genes. The four cytokine genes are located in the region of the long arm of chromosome 5, which is deleted in the 5q- anomaly. The present study provides a basis for further investigations of this disorder. PMID- 2564790 TI - Lymphoid differentiation of the hematopoietic stem cell that reconstitutes total erythropoiesis of a genetically anemic W/Wv mouse. AB - We investigated whether the stem cell that reconstitutes total erythropoiesis of a WBB6F1-W/Wv mouse differentiates into lymphoid lineage. The electrophoretic pattern of hemoglobin was used as a marker of the reconstitution; 3 phosphoglycerate kinase (PGK), an X chromosome-linked enzyme was used as a tool for estimating clonality. We injected 10(5) bone marrow cells of 5-FU treated C57BL/6-Pgk-1b/Pgk-1a female mice, in which each stem cell had either A-type PGK or B-type PGK due to random inactivation of one of two X chromosomes, into genetically anemic (WB x C57BL/6)F1-W/Wv (hereafter WBB6F1-W/Wv) mice that contained only B-type PGK. The recipient WBB6F1-W/Wv mice, in which erythropoiesis was reconstituted with donor cells for a long term, were killed and the PGK patterns of bone marrows, thymus, lymph nodes, and Peyer's patches were examined. A considerable amount of A-type PGK was detected in the lymphoid organs of the WBB6F1-W/Wv mice in which erythrocytes showed only A-type PGK when killed. In contrast, A-type PGK was scarcely detectable in the lymphoid organs of the WBB6F1-W/Wv mice in which erythrocytes showed only B-type PGK when killed. The present results suggest that the hematopoietic stem cells estimated by the erythropoiesis reconstituting assay differentiate into lymphoid lineage and that the long-term erythropoiesis reconstitution assay is useful for detecting the true primitive hematopoietic stem cells. PMID- 2564791 TI - Mixed hematopoietic chimerism after allogeneic transplantation with lymphocyte depleted bone marrow is not associated with a higher incidence of relapse. AB - Using red cell phenotyping, cytogenetic analysis of blood lymphocytes, chromosome studies of bone marrow cells, and restriction fragment length polymorphism (RFLP) studies of peripheral blood cells, we demonstrated a high number of mixed chimeras after allogeneic bone marrow transplantation (BMT). Donor marrow from HLA-A, -B, and -DR identical, mixed lymphocyte culture (MLC) nonreactive siblings was depleted of 98% of lymphocytes using counterflow centrifugation. Thirty-two of 48 recipients (67%) appeared to be mixed chimeras at 6 months after transplantation. The high number of mixed chimeras is probably a result of lymphocyte depletion of the marrow graft and the high sensitivity of red cell phenotyping for the demonstration of minor cell populations (at levels as low as 0.01%). The probability of relapse-free survival from 6 months to 4 years after BMT was 85% for the mixed chimeras and 65% for the complete donor chimeras. We conclude that in this study, mixed chimerism is not associated with a higher incidence of relapse. PMID- 2564793 TI - Risks of dependence on benzodiazepine drugs. PMID- 2564792 TI - Granulocytic differentiation of HL-60 cells is not regulated by DNA de novo methylation. AB - DNA cytosine methylation and transcription of specific genes are inversely correlated. In granulocytic differentiation of HL-60 cells there is a distinct down regulation of the c-myc proto-oncogene expression, which is probably a causal mechanism. With differentiation of HL-60 cells we found no restriction enzyme fragment length polymorphism (RFLP) within the c-myc proto-oncogene, which indicates that there is no loss of regulatory elements (e.g., TATAA boxes within the first exon). Furthermore, we found no de novo methylation in this region. Methylation of other DNA regions, which could influence c-myc expression, is also not necessary for differentiation, as was shown by inhibition of DNA methylase. L Ethionine and S-adenosyl-L-homocysteine are both potent inhibitors of DNA methylase and do not influence proliferation of HL-60 cells, as shown by FACS analysis. PMID- 2564794 TI - Treatment of itching in atopic eczema. PMID- 2564795 TI - Laparoscopy and the management of the impalpable testis. AB - We describe the technique of laparoscopy when seeking to locate an impalpable testis. The procedure was used in 86 boys with unilateral or bilateral undescended testes; of the 103 testes which were sought, 64 were present, 39 were absent and 53 were intra-abdominal. Laparoscopy is useful in the management of the impalpable testis. The information gained from the study is valuable when advising parents of the significance of an impalpable testis. For example, there is a 39% chance it will be absent, a 29% chance that it will be easily placed in the scrotum and a 38% chance that a staged procedure may be necessary. PMID- 2564796 TI - Cryptorchidism. PMID- 2564797 TI - A spider toxin (JSTX) inhibits L-glutamate uptake by rat brain synaptosomes. AB - Joro spider toxin (JSTX), a specific blocker of glutamate receptors, was found to exert a prominent suppressive action on the Na+-dependent binding of L-glutamate to synaptic membranes and on glutamate uptake by synaptosomes in a dose-dependent manner. In contrast, the synthesized 2,4-dihydroxyphenylacetylasparagine (2,4 DHPA-ASN), a common moiety of spider toxins, which has been shown to exhibit almost the same activity as intact JSTX with respect to the inhibition of Na+ independent glutamate binding to its synaptic membrane receptors, shows lower potency in inhibiting Na+-dependent binding and uptake of L-glutamate. From these findings, it is clear that JSTX has the ability to inhibit not only L-glutamate binding to its synaptic membrane receptors but also L-glutamate uptake by synaptosomes, and that polyamines linked to 2,4-DHPA-ASN in the molecule of spider toxins may participate in the inhibition of L-glutamate uptake. PMID- 2564798 TI - Pharmacological characterization of voltage-clamped catfish rod horizontal cell responses to kainic acid. AB - Excitatory amino acid-induced currents were examined in voltage-clamped rod horizontal cells dissociated from the catfish retina. The cells responded to glutamate (GLU) and the GLU analogues kainate (KA), quisqualate (QA), and alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), while N-methyl-D aspartate (NMDA) produced inconsistent responses. Of the effective agonists, only KA produced large, concentration-dependent current responses. While QA, AMPA, GLU, and NMDA were poor agonists, these compounds were able to block rod horizontal cell responses to KA. The rank order potency for this inhibition was: QA greater than AMPA greater than or equal to L-GLU much greater than D-GLU = NMDA. Several excitatory amino acid receptor antagonists were also able to inhibit rod horizontal cell responses to KA. The rank order potency for the inhibition by the compounds tested was: kynurenate greater than cis-piperidine dicarboxylic acid much greater than D,L-alpha-amino-adipate. Comparison of the potency of several ligands to inhibit rod and cone horizontal cell responses to KA suggested similarities in the KA binding sites of both cell types. PMID- 2564799 TI - Effect of phorbol esters on noradrenaline release from cerebral arteries. AB - The effect of phorbol 12-myristate, 13-acetate (PMA), an activator of protein kinase C, on noradrenaline (NA) release from cat cerebral arteries preincubated with [3H]NA was investigated in order to examine the role of that enzyme in this secretion. PMA (3 microM) potentiated tritium release elicited by electrical stimulation (2 Hz, 0.3 ms) without modification of spontaneous secretion, whereas 4 alpha-phorbol 12,13 didecanoate (3 microM), an inactive compound, had no effect. Tritium release evoked by tyramine was not modified by PMA. The electrically evoked tritium secretion was reduced by clonidine (1 microM) or B-HT 920 (0.1 microM), alpha 2-adrenoceptor agonists, and unaffected by yohimbine (1 microM), an alpha 2-adrenoceptor antagonist. The presence of clonidine, B-HT 920 or yohimbine reduced the action of PMA. The facilitatory effect of PMA was not increased by the Ca2+ ionophore A23187 (5 microM). These results suggest that: (1) protein kinase C of perivascular adrenergic nerve endings participates in the exocytotic release of NA; (2) the effects of PMA could be partially due to an interference with prejunctional autoinhibitory alpha 2-adrenoceptors, and (3) the increase of intracellular Ca2+ produced by A23187 appears to be inadequate for potentiating the action of PMA. PMID- 2564801 TI - Patterns of gamma-motoneuron activity in the external intercostal muscles of the cat during respiration. AB - Recordings of gamma-motoneurons from fine filaments of external intercostal nerves were made during spontaneous breathing in the anesthetized cat. The patterns of alpha- and gamma-motoneuron activity varied within different areas of the rib cage. Areas of muscle which were recruited during inspiration received input from both tonically and phasically active gamma-motoneurons. Those areas of the rib cage which were not recruited during inspiration received activity from tonically active gamma-motoneurons. These patterns of gamma-motoneuron activity are in agreement with those suggested from previous recordings of muscle spindle activity. PMID- 2564800 TI - Evidence for excitatory amino acid transmission between mesencephalic nucleus of V afferents and jaw-closer motoneurons in the guinea pig. AB - Previous studies have suggested that monosynaptic transmission between spinal primary afferent fibers and motoneurons is mediated by an excitatory amino acid, most likely glutamate or aspartate. No such comparable studies have been carried out in the trigeminal system. In an attempt to elucidate the neurotransmitter(s) mediating monosynaptic transmission between mesencephalic of V nucleus afferents (Mes V) and trigeminal jaw-closer motoneurons, the effect of iontophoretic application of excitatory amino acid antagonists on the Mes V-induced field potential, recorded in the trigeminal motor nucleus (Mot V), was examined. Application of DL-2-amino-4-phosphonobutyrate (APB) and the broad spectrum amino acid antagonists, kynurenic acid (KYN) and gamma-D-glutamylglycine (DGG), for 3-4 min reversibly reduced the amplitude of the Mes V induced field potential. The effect of APB was much greater than any of the other compounds tested. On the other hand, the specific N-methyl-D-aspartate (NMDA) receptor blocker DL-2-amino 5-phosphonovaleric acid (APV), was without effect on the field potential. Based on current-response curves for each antagonist tested, the order of potency was determined to be APB greater than KYN greater than DGG greater than APV. These antagonists were also compared with respect to their efficacy in blocking individual jaw-closer motoneuron activity induced by iontophoretic application of amino acid receptor excitants glutamate (Glut), aspartate (Asp), kainate (K), and quisqualate (Q). NMDA application was without effects on these motoneurons. The profile of activity of these antagonists on these amino acid excitants was similar to that found in other areas of the CNS by other investigators. KYN and DGG both significantly reduced responses induced by all excitants tested, whereas APB had more modest effects on K and Q excitation and was without effect on Glut and Asp excitations in most cells tested. The data suggest that an excitatory amino acid, activating non-NMDA receptors, mediates some component of synaptic transmission between Mes V afferents and jaw-closer motoneurons. The data is also consistent with the proposal made in other systems that APB blocks synaptic transmission by a mechanism other than postsynaptic receptor blockade. PMID- 2564802 TI - Glycine, like glutamate, microinjected into the nucleus tractus solitarii of rat decreases arterial pressure and heart rate. AB - Glycinergic mechanisms have been implicated in central cardiovascular regulation. However, the inhibitory amino acid's role in the nucleus tractus solitarii (NTS), the site of termination of cardiovascular afferents, has not been clarified. Thus, we sought to determine if the microinjection of glycine into the NTS alters arterial pressure and heart rate. Microinjections of glycine, like glutamate, confined to the NTS decreased arterial pressure and heart rate in a neurally mediated, dose-dependent manner. The glycine antagonist strychnine completely blocked these effects of glycine but did not itself alter arterial pressure or heart rate, or interfere with the baroreceptor reflex. The acute hypotensive, bradycardic response to glycine was followed by a period during which glycine essentially eliminated the cardiovascular responses to the microinjection into the NTS of glutamate, an amino acid reputed to be a transmitter in the baroreceptor reflex arc. These data suggest that glycine is involved in cardiovascular regulation by the NTS but do not support its being an integral transmitter in the baroreceptor reflex. PMID- 2564803 TI - Taxol toxicity. Epithelial necrosis in the gastrointestinal tract associated with polymerized microtubule accumulation and mitotic arrest. AB - Taxol, an antineoplastic agent with a novel mechanism of action, is currently undergoing Phase I trials at The Johns Hopkins Hospital, Baltimore. The authors recently observed striking mitotic arrest associated with epithelial necrosis and ulceration in an esophageal biopsy specimen. The biopsy specimen was taken from a patient who received taxol the day before endoscopy. Review of our autopsy files revealed four other cases in which taxol had been administered. Sections of esophagus, stomach, small intestine, colon, liver, skin, bone marrow, and testes were examined for evidence of mitotic arrest. Mitotic arrest was seen in the two patients who underwent autopsy who received taxol less than 11 days before death, whereas the two autopsy patients who received taxol more than 2 weeks before death did not show these changes. Although mitotic arrest was most prominent in the esophagus, it was also found in the stomach, small intestine, colon, liver, and bone marrow. The mitotic arrest was associated with bundling of intermediate filaments and appeared to be secondary to accumulation of polymerized microtubules. These results suggest that taxol induces a transient mitotic arrest associated with cell necrosis. PMID- 2564804 TI - Paraffin section markers for Reed-Sternberg cells. A comparative study of peanut agglutinin, Leu-M1, LN-2, and Ber-H2. AB - Hodgkin's disease (HD) is sometimes difficult to distinguish from non-Hodgkin's lymphomas, and a reliable marker for Reed-Sternberg and related (R-S) cells in paraffin sections would be useful. Ninety-one cases of HD with PNA, anti-Leu M1, and LN-2, and 90 cases with Ber-H2 were studied. The staining results were evaluated independently. R-S cells stained positively with one or more of the reagents in all cases. PNA staining was positive in 78 cases (85.7%); Leu M1, 63 (69.2%); LN-2, 71 (78.0%); and Ber-H2, 80 cases (88.9%). Positively stained cells were readily recognized in 71 cases (91.0%) of PNA+, 51 (80.9%) of Leu M1+, and 51 (71.8%) of LN-2+ and 71 (88.7%) of Ber-H2+ cases; the cells were found only after careful search in the remaining cases. Sixteen cases of peripheral T-cell lymphoma (large cell type, ten; mixed, five; unclassifiable, one) were also stained. Tumor cells did not stain with PNA or anti-Leu M1 in any of the 16 cases but did stain positively with LN-2 in four and with Ber-H2 in five. Thus, the detection rate of R-S cells was the highest with Ber-H2, closely followed by PNA. PNA, however, stained the largest number of R-S cells per case, and the results were least affected by the type of fixative employed. Staining of peripheral T cell lymphoma appeared to be nil or extremely rare with PNA and Leu M1, whereas it was not uncommon with Ber-H2 and LN-2. In conclusion, to facilitate the detection of R-S cells in paraffin sections, the application of a panel of three markers, PNA, Leu M1, and Ber-H2, appears to be necessary at this point in time. PMID- 2564805 TI - Genetically different cell subpopulations in hydatidiform moles. A study of three cases by RFLP, flow cytometric, cytogenetic, HLA, and morphologic analyses. AB - Most investigators find a good correlation between the morphologic and cytogenetic classification of hydatidiform moles (HM), but exceptions have been noted. We have examined three cases of HM, using chromosome marker analysis on cultured cells, human leukocyte antigen typing on cultured and uncultured tissue, and restriction fragment length polymorphism (RFLP) analysis and flow cytometry on uncultured cells. In one morphologically partial mole, only one cell population (triploid) was found and data obtained by the above-mentioned techniques were concordant. The other two moles, which were classified morphologically as complete, consisted of several cell subpopulations differing in DNA content. In both cases only one cell population was disclosed by cytogenetic investigation. In one case, the cytogenetic analysis indicated that the cultured cells were near triploid with paternal chromosomes exclusively, whereas RFLP analysis showed that maternal X chromosomal alleles were present in the mole. The present findings demonstrate that some HMs contain cellular subpopulations with differing DNA content. One explanation for discordance between cytogenetic and morphologic classification may thus be the detection of only one cell subpopulation when karyotyping. PMID- 2564806 TI - Expression of c-erbB-2 oncoprotein: a prognostic indicator in human breast cancer. AB - Sections of formalin-fixed, paraffin-embedded tissue from 185 primary breast carcinomas were stained immunohistochemically using a polyclonal antibody against the c-erbB-2 oncoprotein. Positive staining, which is known to correlate with gene amplification, was associated with earlier relapse, shorter postrelapse survival, and shorter overall survival. Lymph node, epidermal growth factor receptor, and estrogen receptor status, tumor size, and histological grade also had prognostic significance but, applying multivariate analysis, only lymph node status was a more important predictor of relapse-free and overall survival than staining for the oncoprotein. Positive staining was correlated with negative estrogen receptor status and high histological grade, but there was no association with either lymph node or epidermal growth factor receptor status or tumor size. Expression of the c-erbB-2 oncoprotein appears to be an important independent indicator of prognosis in human breast cancer. PMID- 2564807 TI - Gene regulation and oncogenes. AACR special conference in cancer research. American Association for Cancer Research. PMID- 2564808 TI - Differential modulation of gene induction by glucocorticoids and antiglucocorticoids in rat hepatoma tissue culture cells. AB - Studies of glucocorticoid and antiglucocorticoid induction of tyrosine aminotransferase (TAT) in two rat hepatoma cell lines (Fu5-5 and HTC) are described. These studies revealed several phenomena that are not consistent with the current models of steroid hormone action: (a) TAT induction occurred at glucocorticoid levels below those required for comparable receptor occupancy in Fu5-5, but not in HTC, cells; (b) the ability of antiglucocorticoids to induce TAT is higher in Fu5-5 than in HTC cells; (c) the values of the amount of TAT agonist activity with the antiglucocorticoid dexamethasone 21-mesylate and of log10 of the dexamethasone concentration required for half-maximal induction of TAT were not constant over time but varied in a linear, reciprocal manner. This modulation was seen for several glucocorticoids and antiglucocorticoids at the level of both TAT enzyme and mRNA but not for two other glucocorticoid inducible genes in the same cells. These results, plus the fact that a similar difference in the concentration required for half-maximal TAT induction in Fu5-5 cells was seen for both glucocorticoids and cyclic AMP, argue that the modulation occurs at some point distal to receptor-steroid complex binding to the biologically active nuclear sites but proximal to translation of TAT mRNA. In order to explain these results, it is pointed out that models involving second messengers are entirely appropriate for steroid hormone action. The participation of a modulated trans acting factor in such a model may explain the above results. PMID- 2564809 TI - Oncogene mediated repression of glucocorticoid hormone response elements and glucocorticoid receptor levels. AB - We have previously described the inhibition of glucocorticoid-dependent transcription from the mouse mammary tumor virus long terminal repeat promoter by products of the H-ras and v-mos oncogenes. We have studied the effects of conditional oncogenes on expression of glucocorticoid-dependent indicator genes. Expression of the glucocorticoid-dependent transcription of the tyrosine aminotransferase gene was monitored in FTO-2B rat hepatoma cells during Mr 21,000 protein (p21) H-ras induction. A strong transcriptional repression of the tyrosine aminotransferase gene followed p21 H-ras expression. The sequences in a glucocorticoid-dependent promoter which are responsible for the oncogene-mediated repression could be localized to the glucocorticoid response element; a construct in which a 15-base pair glucocorticoid response element was inserted 5' of the thymidine kinase promoter exhibited the oncogene-mediated repression of transcription. We observed a strong repression of glucocorticoid-dependent promoters and promoter constructs not only in the presence of p21 H-ras and p37 v mos but also with p60 v-src. p57 v-myc, however, had no effect. Oncogene expression is not a sufficient prerequisite for an initial repression of glucocorticoid hormone-dependent gene transcription, since even in the presence of constitutively high levels of oncogene product a transient stimulation of glucocorticoid-dependent gene expression was found. Protein synthesis inhibition experiments revealed that no hormonally induced cellular protein is needed for the oncogene-mediated repression. It seemed reasonable that this phenomenon might reflect oncogene effects on the glucocorticoid receptor. We, therefore, made measurements of the glucocorticoid receptor protein. In the presence of glucocorticoid hormone the receptor translocated rapidly from the cytoplasm to the nucleus. In normal NIH 3T3 cells, after 24-h treatments the nuclear receptor levels had declined to about 50% of those determined at 2 h and in the presence of p21 H-ras they declined to 15%. The levels of cytoplasmic receptor were not affected by p21 H-ras expression. PMID- 2564810 TI - Bilateral subdural empyema due to Salmonella enteritidis in an infant. AB - A case is reported of an infant with bilateral subdural empyema due to Salmonella enteritidis. This Salmonella species is extremely uncommon as a pathogen in a focal infection in the central nervous system. Treatment by early surgical drainage, followed by systemic antibiotics and subdural irrigation, was successful. This is the first case reported of infantile subdural empyema due to S. enteritidis. PMID- 2564811 TI - Amino-terminal sequences of prosomatostatin direct intracellular targeting but not processing specificity. AB - Rat preprosomatostatin (rPPSS) is processed to two bioactive peptides, somatostatin-14 and somatostatin-28. In anglerfish islets, the two peptides are synthesized by distinct cell types and are derived from different precursors, anglerfish preprosomatostatin-1 (a(I)PPSS) and anglerfish preprosomatostatin-2 (a(II)PPSS). To determine the basis of the differential processing, we introduced a(I)PPSS or a(II)PPSS expression vectors into mammalian endocrine cell lines that can accomplish both patterns of processing. Both precursors were processed identically, indicating that cellular factors must determine the processing pattern. Although similar processing sites are present in both precursors, high levels of unprocessed anglerfish prosomatostatin-2 were secreted constitutively from the transfected cells. A hybrid protein containing the leader sequence and a portion of the pro-region of rPPSS fused to the carboxy-terminal third of a(II)PPSS was processed and secreted via a regulated pathway. We conclude that the amino-terminal 78 residues of rPPSS contain sufficient information to correct the targeting deficiency of a(II)PPSS in mammalian endocrine cell lines. PMID- 2564812 TI - Determination of axial polarity in the vertebrate embryo: homeodomain proteins and homeogenetic induction. PMID- 2564813 TI - Involvement of the Xenopus homeobox gene Xhox3 in pattern formation along the anterior-posterior axis. AB - The Xenopus homeobox gene Xhox3 shows a graded expression in the axial mesoderm, with the highest concentration in the posterior end of frog gastrula and neurula embryos. To investigate the function of the Xhox3 gene, synthetic Xhox3 mRNA was injected into different regions of developing embryos. In particular, Xhox3 was supplied in excess to anterior cells, which normally have the lowest levels of Xhox3 RNA. The results show that injection of Xhox3, but not control, mRNA into prospective anterior regions of developing embryos produces a series of graded axial defects. The injected embryos gastrulate normally but fail to form anterior (head) structures. Our findings suggest that Xhox3 is involved in establishing anterior-posterior cell identities during pattern formation of the axial mesoderm in early embryonic development. PMID- 2564814 TI - Glutathione and lipid peroxide levels in rat liver following administration of methapyrilene and analogs. AB - The possibility was examined that the induction of tumors in rat liver by feeding methapyrilene, which is not mutagenic, is related to effects on glutathione levels and lipid peroxidation. Fischer 344 rats were given single-dose and multiple-dose treatments with the anti-histamine methapyrilene (MP), which is carcinogenic in rats, and with two non-carcinogenic analogs, methafurylene (MF) and thenyldiamine (TD) and the effects on malonaldehyde (MDA) formation and glutathione (GSH) levels in the liver were investigated. After a single dose, MDA levels were increased at 6 h by MF and TD and at 24 h by MP. MDA levels returned to normal after 30 h with MP and MF, but not with TD. Levels of MDA (and other TBA-reactive products) after four daily treatments were most elevated by TD, less elevated by MP, and were lowered by MF. Forty-two hours following treatment with both MP and MF, MDA levels had returned to normal, but in TD-treated animals MDA remained high. GSH levels were highest after MF and MP, and remained high at 42 h, but TD induced only a small increase. There appears to be increased lipid peroxidation in the liver as a result of treatment of rats with MP, MF and TD. The greater response induced by TD, as well as the increased liver GSH levels after repeated administration of all three drugs indicate that lipid peroxidation in rat liver is not a particular effect related to the liver carcinogen methapyrilene. PMID- 2564815 TI - gamma-Glutamyltranspeptidase-negative phenotypic property of preneoplastic and neoplastic liver lesions induced by ciprofibrate does not change following 2 acetylaminofluorene administration. AB - Preneoplastic and neoplastic lesions induced by peroxisome proliferators in the livers of rats and mice do not express gamma-glutamyltranspeptidase (GGT). Previous studies have shown that the absence of GGT is not due to the toxic effect of peroxisome proliferators or due to the presence of inactive enzyme. The present experiment was designed to examine whether the GGT-negative property of these lesions is stable and irreversible or whether these lesions can be modulated to express GGT by 2-acetylaminofluorene (AAF). Hepatic lesions were induced in F-344 rats by feeding ciprofibrate (0.025%) in diet for 60 or 84 weeks. These rats were then administered AAF in diet (0.02%) for 5 weeks and the altered areas (AAs), neoplastic nodules (NNs) and hepatocellular carcinomas were analyzed for the expression of GGT. Ninety per cent of carcinomas, 90-100% of NNs and greater than 60% AAs were negative for GGT following AAF treatment. These results are very similar to the phenotypic features observed in the livers of rats treated with ciprofibrate alone. The results of this study suggest that the GGT-negative property of ciprofibrate-induced lesions is stable and not modulatable by AAF. PMID- 2564816 TI - DNA probes for carrier identification in Duchenne muscular dystrophy. PMID- 2564817 TI - Enzymic urea assay: a new colorimetric method based on hydrogen peroxide measurement. AB - We describe a new enzymic colorimetric method in which urea is measured in serum by use of a single reagent mixture. Ammonia produced by urea hydrolysis, catalyzed by urease, reacts with glutamate and ATP in the presence of glutamine synthetase. The ADP so produced is assayed in reactions catalyzed sequentially by pyruvate kinase and pyruvate oxidase in a system that generates hydrogen peroxide. The hydrogen peroxide is measured at 500 or 550 nm in a reaction catalyzed by horseradish peroxidase, with phenol/4-aminophenazone as the chromogen. The reaction is complete in 15 min at 37 degrees C. The standard curve is linear up to a urea concentration of 40 mmol/L. Precision is good; CVs ranged from 2.5% to 3.1%. Results by the present method compared well with those by a candidate Reference Method and are not subject to interferences from commonly used drugs and anticoagulants. PMID- 2564818 TI - Use of DNA probes in detecting carriers of Duchenne muscular dystrophy: selected case studies. AB - Detection of Duchenne muscular dystrophy carriers by genetic analysis is illustrated by four case studies. The technique is most useful in obligate families, in excluding carrier status in isolated cases, and in families in which the affected child demonstrates a molecular deletion. A major limitation of this technique is that the accuracy of carrier status in isolated (i.e., no family history) cases is limited by the probability that the affected child may represent a new mutation. To improve the carrier risk estimate generated by the DNA data, particularly in isolated cases, we suggest that measuring creatine kinase activities in the serum and performing the genetic analysis on the nonaffected males may be helpful. PMID- 2564819 TI - Somatostatin analogues in diabetes mellitus. AB - Growth hormone (GH) has long been considered to have importance in diabetes. With poor control in Type 1 diabetes GH levels are high and may aggravate poor metabolic control. Pharmacological suppression of GH release at this stage might reverse the metabolic changes, with the possible added benefit of lower plasma insulin concentrations. Diabetic patients with life-long GH deficiency rarely develop retinopathy, while pituitary ablation in patients with retinopathy often leads to improvement. Growth hormone release inhibiting factor, somatostatin, has a short plasma half-life, and multiple effects on the endocrine system and on the gastrointestinal tract, making it unsuitable for clinical use as a GH suppressant. Long-acting analogues have a long half-life, but remain non-specific in their effects. In Type 2 diabetes the analogue Octreotide suppresses insulin and glucagon release, leaving glucose levels either unchanged or somewhat elevated. Gastrointestinal side-effects have been common, but may diminish with long-term treatment. In Type 1 diabetes insulin requirement is decreased by Octreotide, but as in Type 2 diabetes GH suppression has been observed consistently only when the drug was given at bed-time. The decrease in insulin requirement may reflect suppression of glucagon release and/or gut effects. Amelioration of the 'dawn phenomenon' has not proved possible, and hypoglycaemia has proved a particular problem with Octreotide given subcutaneously at night. The lack of effective GH suppression (particularly in patients with proliferative retinopathy), lack of specificity, and the gut and hypoglycaemic side-effects, argue strongly against a clinical role for the current somatostatin analogues in diabetes mellitus. PMID- 2564820 TI - A most memorable case. PMID- 2564821 TI - Participation of nicotinic receptor in hormone release from isolated rat islets of Langerhans. AB - Pancreatic hormone release is generally thought to be regulated through adrenergic as well as muscarinic receptors. We have previously observed possible nicotinic involvement in insulin release. In the present study, we incubated isolated rat islets for 60 min with various concentrations of atropine (a muscarinic receptor blocker), alpha-bungarotoxin (alpha-Btx, a nicotinic receptor blocker), and anti-acetylcholine receptor antibody (IgG) (anti-Ach.R.Ab) obtained from a patient with myasthenia gravis. Atropine suppressed insulin release, and alpha-Btx and anti-Ach.R.Ab potentiated it; atropine did not suppress glucagon release, while alpha-Btx and anti-Ach.R.Ab raised it. None of these agents influenced somatostatin release. These observations suggest that muscarinic as well as nicotinic receptors influence insulin release, as nicotinic receptors do glucagon release. Neither nicotinic nor muscarinic receptors seem to regulate somatostatin release. PMID- 2564822 TI - [New data on tardive dyskinesias]. PMID- 2564823 TI - Long-term reduction in sudden deaths after a multifactorial intervention programme in patients with myocardial infarction: 10-year results of a controlled investigation. AB - Three-hundred and seventy-five unselected patients below 65 years of age and with acute myocardial infarction participated in a controlled investigation aimed at studying the effects of a multifactorial intervention programme on morbidity, mortality and risk factor control. After ten years' follow-up the significantly lower sudden death and coronary mortality observed three years after myocardial infarction still persisted in the intervention group (188 patients) compared with the control group (187 patients). The incidence of sudden death in the intervention group was 12.8% compared with 23.0% in the controls (P = 0.01). The incidence of coronary mortality was 35.1% and 47.1%, respectively (P = 0.02). No significant difference was found in the number of patients with clinical non fatal reinfarctions (25.6% and 19.3%, respectively). During the first year, when the mortality difference was most marked, the use of beta blockers was not significantly different between the groups. The results suggest that with a multifactorial intervention programme which starts early after the infarction and lasts for years a significant long-term reduction in sudden deaths and coronary mortality can be attained. PMID- 2564824 TI - Antibody 12-15 cross-reacts with mouse Fc gamma receptors and CD2: study of thymus expression, genetic polymorphism and biosynthesis of the CD2 protein. AB - Previously we described a monoclonal antibody (mAb 12-15) that reacted with murine Fc receptor proteins (beta 1, beta 2 and alpha) and an undefined molecule of 37 kDa (beta 3) on certain types of cells. Here we present serological and biochemical evidence that the beta 3 chain is expressed on mouse thymocytes and that it is identical to the CD2 antigen. By immunofluorescence staining and cytofluorographic analysis greater than 95% of thymocytes were positive. Brightly staining cells coincided with cortisone-resistant thymocytes suggesting that mature thymocytes expressed higher levels of the antigen. Biosynthetic labeling of DBA/2 thymocytes with [35S]methionine showed that the size of the CD2 precursor molecule was 43 kDa which was processed to approximately 55-65 kDa in the mature molecule. mAb 12-15 was also reactive with the tunicamycin-treated form of the CD2 antigen suggesting that the cross-reactive epitope was of protein nature. Comparison of different mouse strains indicated that two molecular forms of CD2 exist. On BALB/c thymocytes, the relative mass of the native molecule was approximately 60-70 kDa (CD2.1) and slightly larger than in DBA/2 (CD2.2). Following endoglycosidase F treatment both proteins still showed a slight difference in electrophoretic mobility. Several inbred mouse strains were analyzed for expression of CD2 forms. When mAb 12-15 was used in cytotoxic T lymphocyte inhibition experiments using specific CTL and tumor target cells it was found that the antibody could specifically inhibit CTL-mediated lysis presumably by interfering with CD2 function. PMID- 2564825 TI - Malaria transmission-enhancing activity in mosquitoes by mammalian host anti sporozoite antibodies. PMID- 2564829 TI - [Work organization of the occupational pathology department of a provincial hospital]. PMID- 2564830 TI - Etilefrine and amezinium reduce uterine blood flow of pregnant guinea pigs. AB - Etilefrine and Amezinium are used during pregnancy to prevent hypotension and fetal growth retardation. The effect of these drugs on uterine blood flow (UBF), uterine vascular resistance (UVR) and fetal growth are, however, unknown. 31 guinea pigs were injected daily with Etilefrine (3 X 0.14 mg/kg) and Amezinium (0.14 mg/kg) from day 30 to day 60. Uterine blood flow was measured with radioactive-labeled microspheres. Uterine vascular resistance was calculated from arterial blood pressure and uterine blood flow. 10 guinea pigs treated with 0.9% sodium chloride served as controls. As a response to Etilefrine and Amezinium, UBF fell by 68 and 48%, respectively, accompanied by an increase in UVR. The fetal weight decreased only slightly. It is concluded that long-term application of anti-hypotensive drugs may be hazardous to uterine blood flow and fetal growth. PMID- 2564831 TI - Variable receptor affinity hypothesis. AB - Measurements of the contractile response to norepinephrine (NE) of a variety of arteries of three mammalian species that are commonly used in the laboratory provide evidence that tissue sensitivity and affinity of the alpha 1-adrenoceptor for NE covary over a range of several orders of magnitude. The quantitative relationship suggests that variation in affinity can, to a great extent, account for the variation in sensitivity found in a number of circumstances. Furthermore, it is argued that the variation in affinity appears to be continuous and thus does not provide a basis for receptor type subdivision. There is also evidence that adrenergic antagonist affinity can vary significantly in tissues. The factors that might account for this variation include differences in receptor chemical structure, in the local membrane microenvironment, and in a number of intracellular processes. A hypothesis of variable receptor affinity has been proposed. If it is correct, then variation in receptor affinity is an important functionally relevant variable that could account for selectivity of tissue responses to circulating hormones and may represent a mechanism of change in the intact organism and in disease. PMID- 2564832 TI - Are antacids cytoprotective? PMID- 2564833 TI - Duodenal and antral mucosal prostaglandin E2 synthesis in a study of normal subjects and all stages of duodenal ulcer disease treated by H2 receptor antagonists. AB - We tested the hypothesis that the production of prostaglandin E2 (PGE2) is impaired in duodenal ulcer disease and affected by treatment and healing. This was investigated by a study of maximal PGE2 synthesis rates in duodenal and antral mucosal biopsies obtained at endoscopy. The patients were divided into three groups. Group (a): endoscopically normal controls (n = 56); group (b): treatment controls (non-DU disease: gastric ulcer or oesophagitis treated by histamine H2 receptor antagonists) (n = 41); and group (c): patients with DU disease (n = 183) further subdivided into group (c1) active ulcer not on treatment (n = 47), (c2) treated active ulcer (n = 35), (c3) healed ulcer on treatment (n = 86), and (c4) healed ulcer not on treatment (n = 15). Group (a) synthesised (mean (SD] 106.6 (39.0) pg PGE2/mg wt of tissue from the duodenal bulb and 129.9 (56.9) from the second part of the duodenum. No difference was found between group (a) and (b) at either site. Group (c1) ulcer rim made 49.8 (22.7) and at all stages ulcer rim and scar made less than the control duodenal bulb (p less than 0.02). Uninvolved duodenal bulb form groups (c1) (63.4 (31.0], (c2) (83.6 (38.5], and (c3) (81.5 (31.1], however, also made significantly less than controls (p less than 0.02) and a similar though non-significant trend was seen in group (c4). Biopsies from the second part of the duodenum did not synthesise significantly less than the control group but a similar trend was noticed at each stage of ulcer treatment. Biopsies of control antrum synthesised 124.5 (32.2) but only 93.7 (44.2) in group (cl) (p < 0.005). All stages of duodenal ulcer healing were associated with a decreased capacity to synthesise the major prostaglandin PGE2 at the ulcer site and the uninvolved duodenal bulb and, in acute untreated duodenal ulcer, the uninvolved antrum. This decreased capacity may be the consequence of the disease process itself and not secondary to the treatment, indicating a basic pathophysiological abnormality which may explain the characteristic tendency of the disease to relapse. PMID- 2564834 TI - Somatostatin in the treatment of acute pancreatitis: a prospective randomised controlled trial. AB - A prospective study was carried out to evaluate the efficacy of somatostatin in the treatment of acute pancreatitis. Seventy one patients were randomised to control (h = 36), or to the somatostatin group (h = 35) who received somatostatin 100 micrograms/h after a 250 microgram bolus for the first two days. The following were compared in the two groups on admission and two days later: laboratory tests of prognostic significance, severity of pancreatitis, and also morbidity and mortality. Of the nine laboratory tests compared, the white blood cell count, lactate dehydrogenase, and urea concentrations were significantly lower in the somatostatin group two days after admission. Severity of pancreatitis after hospitalisation increased in fewer patients given somatostatin (NS). There was a trend toward fewer complications, especially local, in the somatostatin group. Mortality in both groups was low. Somatostatin appeared to reduce the local complications of acute pancreatitis. A larger trial is necessary to show its beneficial effect conclusively. PMID- 2564835 TI - A simple gas chromatographic identification and determination of 11 CNS stimulants in biological samples. Application on a fatality involving phendimetrazine. AB - A method is presented for the simultaneous identification and quantification of several CNS stimulants, including amphetamine in plasma and urine by GC/FID using mephentermine as an internal standard. No derivation is necessary and after a single alkaline extraction, GC analysis for the 11 compounds tested is achieved in 23 min. The lower limit of detectability was found to be 4 ng/ml for amphetamine in plasma. This method is sensitive, reproducible, selective and applicable in forensic and clinical toxicological analyses. Toxicological findings, after a fatality involving phendimetrazine are presented as an application of the procedure. PMID- 2564836 TI - Gene of X-chromosomal congenital stationary night blindness is closely linked to DXS7 on Xp. AB - Congenital stationary night blindness is characterized disturbed or absent night vision that is always present at or shortly after birth and nonprogressive. The X linked form of the disease (CSNBX; McKusick catalog no. 31050) differs from the autosomal types in that the former is frequently associated with myopia. X chromosome-specific polymorphic DNA markers were used to carry out linkage analysis in three European families segregating for CSNBX. Close linkage without recombination was found between the disease locus and the anonymous locus DXS7, mapped to Xp11.3, assigning the mutation to the proximal short arm of the X chromosome. Linkage data obtained with markers flanking DXS7 provided further support for this localization of the gene locus. Thus, in addition to retinitis pigmentosa and Norrie disease, CSNBX represents the third well-known hereditary eye disease the locus of which is mapped on the proximal Xp and closely linked to DXS7. PMID- 2564837 TI - Rapid isolation of moderate and highly polymorphic DNA fragments mapping close to WT (Wilms' tumour) and AN2 (aniridia) on chromosome 11. AB - Genes implicated in the development of Wilms' tumour (WT) and aniridia (AN2) have been localised to a sub-region of band p13 on chromosome 11 by molecular and cytogenetic characterisation of WAGR syndrome patients carrying variable constitutional deletions. Polymorphic markers for the region would be valuable for linkage analysis in the familial forms of both Wilms' tumour and aniridia, as well as for studying somatic rearrangements of chromosome 11 in a variety of tumour types. Here we describe the isolation and characterisation of three frequently polymorphic arbitrary DNA fragments that map proximal to the AN2 and WT loci. PMID- 2564838 TI - Study of a family with a fragile site of the X chromosome at Xq27-28 without mental retardation. AB - The fragile site Xq27-28 was observed in several individuals of a large family. It is expressed at a high frequency among the carrier females, even as adults, and in one clinically normal male. None of the members of this family is affected with the mental retardation normally linked to this fragile site. Cytogenetic and flanking DNA marker polymorphism studies suggest a possible dissociation between the fragile site and clinical expression of the disease. PMID- 2564839 TI - DNA haplotype analysis at the phenylalanine hydroxylase locus in the Turkish population. AB - Thirty-nine Turkish phenylketonuria (PKU) families were investigated for their DNA haplotypes at the phenylalanine hydroxylase (PAH) locus. There was a threefold higher incidence of consanguinity in the population studied compared with the general Turkish population. The PAH DNA haplotype 6 was found to be almost exclusively associated not only with the mutant PAH genes but also with the classic phenotype in 39% of the Turkish patients. This haplotype was of not importance in northern European populations. The two DNA haplotypes (1 and 4) that were almost equally frequent among the normal and the mutant PAH genes in northern European populations show virtually the same distribution in Turkish individuals. In all populations studied, these haplotypes are associated with different phenotypes. PMID- 2564840 TI - Confirmation and refinement of the localisation of the c-MEL locus on chromosome 19 by physical and genetic mapping. AB - The human gene locus c-MEL was identified following transfection of genomic DNA from the human melanoma cell line NK14; it has previously been assigned to chromosome 19 (p13.2-q13.2) by analysis of somatic cell hybrids. We have further refined the position of this gene to the proximal region of 19p (cen-p13.2), using cell hybrids containing only fragments of human chromosome 19. We have confirmed this physical localisation by linkage analysis with a recently described restriction fragment length polymorphism for the c-MEL gene, and mapped the locus within the region of the low density lipoprotein receptor gene (LDLR) (Lod 4.43, theta = 0.10) and the anonymous marker D19S11 (13.1.25) (Lod 9.33, theta = 0). This gene thus maps to a region of chromosome 19 involved in karyotypic abnormalities in a variety of malignancies including melanomas and leukaemias. PMID- 2564841 TI - A new polymorphic DNA probe pS43 derived from a flow sorted library is assigned to human chromosome 20q13. AB - Human chromosome 20 has not been sufficiently mapped, as only four DNA probes detecting RFLPs and 18 genes have been assigned to this chromosome. Using a chromosome 20-specific library to isolate and characterize new low-copy DNA probes, we found a new polymorphic DNA probe (pS43), which is assigned to human chromosome 20q13. PMID- 2564842 TI - Restricted expression of CD2 among subsets of sheep thymocytes and T lymphocytes. AB - A monoclonal antibody (mAb) generated against sheep T-cell blasts, called I/35 A, blocks sheep autologous E rosetting and competes with purified T11 target structure (TS), the sheep form of LFA3, for binding sites on the sheep T-cell surface. Immunoprecipitation from lysates of surface iodinated sheep T cells identifies the cell surface molecule recognized by mAb I/35 A as a single chain polypeptide migrating as a diffuse band of MW 55,000. From its binding properties and the biochemical nature of the target antigen, we conclude that mAb I/35 A is directed at sheep CD2. This finding makes sheep the first animal model in which the CD2-LFA3 (T11TS) system is defined by mAbs to both receptor and ligand. When analysed by two-colour flow cytometry and by immunohistochemistry, the cellular expression of CD2 in sheep differs significantly to that reported in humans. In peripheral blood, CD2 is found exclusively on CD4+8- and CD4-8+ T cells, while the third, CD4-8- (predominantly SBU-T19+) subset of sheep T cells (around 20% in peripheral blood) is CD2-. In thymus, only low to moderate levels of CD2 expression occurs on 80% of cells. Among these, medullary 'single positive' thymocytes express the highest level of CD2, whereas the CD4-8- 'double negative' population (which in contrast to peripheral CD4-8- T cells contains only very few SBU-T19+ cells) consists of CD2- and weakly positive cells. In peripheral lymphoid organs, CD2+ lymphocytes occur in the T-cell regions of spleen, lymph nodes and jejunal Peyer's patches (JPP). Tissue macrophages found in B-cell follicles of lymph nodes and JPP are also CD2+. The implications of these findings are discussed in terms of the role CD2 plays in the proliferation of immature thymocytes and of the possible importance of CD2/LFA3 interactions in lymphocyte recirculation. PMID- 2564843 TI - Binding of monoclonal antibody to CD16 causes calcium mobilization in large granular lymphocytes but inhibits NK killing. AB - A monoclonal antibody (mAb), CLB/FcR gran I, reactive with the CD16 Fc receptor (FcRlo/FcRIII) of human cells, leads to calcium mobilization in large granular lymphocytes (LGL) but not in granulocytes. Identical responses are obtained with F(ab')2 fragments of this antibody, indicating that the response is independent of Fc-FcR binding, and that bivalent cross-linking of this receptor is adequate for optimal calcium mobilization. The calcium response was greater in CD3- LGL compared to CD3+ LGL, although the response was augmented in the latter cells by prior rosetting with sheep red blood cells (SRBC). Calcium mobilization in CD3- LGL induced by CLB/FcR gran I is associated with inhibition of natural killer cell (NK) killing, and inhibition of the enhanced NK killing induced by the anti CD2 low-density monoclonal antibody, 9.1. This supports the view that the NK enhancing activity of 9.1 is due to simultaneous binding to CD2 and CD16, and may in fact be transduced through the CD16 molecule. The variable reported effects of anti-CD16 antibodies on NK killing are likely to reflect the epitope bound rather than the isotype of antibody used, since F(ab')2 fragments of CLB/FcR gran I also inhibit NK killing. PMID- 2564844 TI - Remarkable sequence conservation of the HLA-DQB2 locus (DX beta) within the highly polymorphic DQ subregion of the human MHC. AB - The HLA-D region of the major histocompatibility complex (MHC) is characterized by a remarkable diversity. Most of the HLA class II genes are highly polymorphic, and in addition, the number and organization of individual loci in that region varies in different haplotypes. This extensive allelic polymorphism of immune response genes has well-known functional implications. Within the HLA-D region, two loci, DQA2 and DQB2 (formerly called DX alpha and DX beta), represent a very special case: the detailed structure of these two genes is entirely compatible with expression, yet their expression has never been demonstrated in any tissue. Consequently, there exists no known corresponding protein product. Pseudogenes are known to accumulate mutations, as observed for instance in the case of HLA DPA2,-DPB2, or -DRB2 genes. We have therefore investigated the extent of DQ2 genes' polymorphism by DNA sequence comparison and by oligonucleotide hybridization across a large number of different haplotypes, and compared it with other genes in the HLA-D region. We show here that, contrary to the adjacent DQ1 genes, DQ2 genes exhibit little and possibly no polymorphism. This conservation of DQ2 genes in many haplotypes indicates that the DQ1-DQ2 duplication event must have preceeded the extensive diversification of DQ1 genes and raises the puzzling question of why DQ2 genes have remained nonpolymorphic. This suggests that either these genes correspond to an unusually invariant region of the MHC or they are under a strong selective pressure for the conservation of the amino acid sequence of a putative DQ2 gene product. The latter would imply that the HLA-DQ2 genes are expressed into a protein product endowed with essential functional properties. PMID- 2564845 TI - Definition of rabbit class I and class II MHC gene haplotypes using molecular typing procedures. PMID- 2564846 TI - Polymorphism of three HLA-DR3-bearing major histocompatibility complex extended haplotypes. PMID- 2564847 TI - Assembly of mutant pilins in Pseudomonas aeruginosa: formation of pili composed of heterologous subunits. AB - Recently, we reported the degree of N-terminal processing within the cytoplasmic membranes of three mutant pilins from Pseudomonas aeruginosa PAK with respect to leader peptide removal and the methylation of the N-terminal phenylalanine (B. L. Pasloske and W. Paranchych, Mol. Microbiol. 2:489-495, 1988). The results of those experiments showed that the deletion of 4 or 8 amino acids within the highly conserved N terminus greatly inhibited leader peptide removal. On the other hand, the mutation of the glutamate at position 5 to a lysine permitted leader peptide cleavage but inhibited transmethylase activity. In this report, we have examined the effects of these mutant pilins upon pilus assembly in a P. aeruginosa PAO host with or without the chromosomally encoded pilin gene present. Pilins with deletions of 4 or 8 amino acids in the N-terminal region were not incorporated into pili. Interestingly, pilin subunits containing the glutamate-to lysine mutation were incorporated into compound pili together with PAO wild-type subunits. However, the mutant pilins were unable to polymerize as a homopolymer. When wild-type PAK and PAO pilin subunits were expressed in the same bacterial strain, the pilin subunits assembled into homopolymeric pili containing one or the other type of subunit. PMID- 2564848 TI - Ammonia assimilation pathways in nitrogen-fixing Clostridium kluyverii and Clostridium butyricum. AB - Pathways of ammonia assimilation into glutamic acid were investigated in ammonia grown and N2-fixing Clostridium kluyverii and Clostridium butyricum by measuring the specific activities of glutamate dehydrogenase, glutamine synthetase, and glutamate synthase. C. kluyverii had NADPH-glutamate dehydrogenase with a Km of 12.0 mM for NH4+. The glutamate dehydrogenase pathway played an important role in ammonia assimilation in ammonia-grown cells but was found to play a minor role relative to that of the glutamine synthetase/NADPH-glutamate synthase pathway in nitrogen-fixing cells when the intracellular NH4+ concentration and the low affinity of the enzyme for NH4+ were taken into account. In C. butyricum grown on glucose-salt medium with ammonia or N2 as the nitrogen source, glutamate dehydrogenase activity was undetectable, and the glutamine synthetase/NADH glutamate synthase pathway was the predominant pathway of ammonia assimilation. Under these growth conditions, C. butyricum also lacked the activity of glucose-6 phosphate dehydrogenase, which catalyzes the regeneration of NADPH from NADP+. However, high activities of glucose-6-phosphate dehydrogenase as well as of NADPH glutamate dehydrogenase with a Km of 2.8 mM for NH4+ were present in C. butyricum after growth on complex nitrogen and carbon sources. The ammonia-assimilating pathway of N2-fixing C. butyricum, which differs from that of the previously studied Bacillus polymyxa and Bacillus macerans, is discussed in relation to possible effects of the availability of ATP and of NADPH on ammonia-assimilating pathways. PMID- 2564849 TI - The membrane form of guanylate cyclase. Homology with a subunit of the cytoplasmic form of the enzyme. AB - A cDNA clone for the membrane form of guanylate cyclase has been isolated from the testis of the sea urchin Strongylocentrotus purpuratus. An open reading frame predicts a protein of 1125 amino acids including an apparent signal peptide of 21 residues; a single transmembrane domain of 25 amino acids divided the mature protein into an amino-terminal, extracellular domain of 485 amino acids and a carboxyl domain of 594 intracellular amino acids. Three potential Asn-linked glycosylation sites were present in the proposed extracellular domain. The deduced protein sequence was homologous to the protein kinase family and contained limited but significant regions of identity with a low molecular weight atrial natriuretic peptide receptor. The carboxyl region (202 amino acids) was 42% identical with a subunit of the cytoplasmic form of guanylate cyclase recently cloned from bovine lung (Koesling, D., Herz, J., Gausepohl, H., Niroomand, F., Hinsch, K.-D., Mulsch, A., Bohme, E., Schultz, G., and Frank, R. (1988) FEBS Lett. 239, 29-34). Therefore, the membrane form of guanylate cyclase is a member of an apparently large family of proteins that includes the low molecular weight atrial natriuretic peptide receptor, the soluble form of guanylate cyclase and protein kinases. PMID- 2564850 TI - Purification and characterization of a 43-kDa transcription factor required for rat somatostatin gene expression. AB - A 43-kDa DNA binding protein which recognizes the TGACGTCA element of the rat somatostatin promoter has been purified from rat brain. Purification of the protein involved initial separation of three sequence-specific binding activities, b1-b3, from each other using DEAE-Sepharose chromatography. The protein corresponding to the b2 complex was further purified to apparent homogeneity by two cycles of sequence-specific DNA affinity chromatography, yielding a single species with an apparent mass of 43,000 daltons on a silver stained polyacrylamide gel. Sequence-specific DNA binding of this purified protein was demonstrated by Southwestern blotting, renaturation, and DNase I footprinting studies. The 43-kDa protein was phosphorylated on serine residue(s) by the catalytic subunit of cAMP-dependent protein kinase, as shown by phosphoamino acid analysis. Furthermore, the purified protein specifically stimulated transcription from the rat somatostatin promoter in an in vitro transcription system. These results indicate that this 43-kDa protein is a transcription factor required for somatostatin gene expression. PMID- 2564852 TI - Nonrandom X chromosome DNA methylation patterns in hemophiliac females. AB - Molecular X chromosome inactivation analysis was used to characterize three females (and their families) with severe hemophilia. First, the maternal and paternal X chromosomes were distinguished by restriction fragment length polymorphisms (RFLPs). Second, the patterns of methylation of X chromosome genes using methylation-sensitive restriction endonucleases were determined. Of the six X chromosome probes tested, only the phosphoglycerol-kinase (PGK) and hypoxanthine-phosphoribosyl-transferase (HPRT) clones were informative, indicating that other X chromosome probes are not useful for X inactivation analysis. After digestion with Hpa II or Hha I, the hybridization intensity of the RFLPs of all three mothers and an unaffected sister were diminished by 50%, consistent with random X chromosome inactivation. The methylation patterns of the X chromosomes of the affected females, however, were clearly nonrandom. Depending upon the probe and the patient, HPRT and PGK sequences were either completely methylated or unmethylated. These findings are extremely suggestive that nonrandom X chromosome inactivation (lyonization) is the basis for severe hemophilia in these females. PMID- 2564851 TI - Human myeloid plasma membrane glycoprotein CD13 (gp150) is identical to aminopeptidase N. AB - To determine the primary structure of CD13, a 150-kD cell surface glycoprotein originally identified on subsets of normal and malignant human myeloid cells, we isolated the complete sequences encoding the polypeptide in overlapping complementary DNA (cDNA) clones. The authenticity of our cDNA clones was demonstrated by the ability of the coding sequences, subcloned in a retroviral expression vector, to mediate expression of bona fide CD13 molecules at the surface of transfected mouse fibroblasts. The nucleotide sequence predicts a 967 amino acid integral membrane protein with a single, 24 amino acid hydrophobic segment near the amino terminus. Amino-terminal protein sequence analysis of CD13 molecules indicated that the hydrophobic segment is not cleaved, but rather serves as both a signal for membrane insertion and as a stable membrane-spanning segment. The remainder of the molecule consists of a large extracellular carboxyterminal domain, which contains a pentapeptide consensus sequence characteristic of members of the zinc-binding metalloprotease superfamily. Sequence comparisons with known enzymes of this class revealed that CD13 is identical to aminopeptidase N, a membrane-bound glycoprotein thought to be involved in the metabolism of regulatory peptides by diverse cell types, including small intestinal and renal tubular epithelial cells, macrophages, granulocytes, and synaptic membranes prepared from cells of the central nervous system. PMID- 2564853 TI - Ultrastructural localization of tyrosine hydroxylase-like immunoreactivity in the rat hippocampal formation. AB - The light and electron microscopic localization of antigenic sites for a polyclonal antiserum directed against the catecholamine synthesizing enzyme, tyrosine hydroxylase (TH), was examined in the hippocampal formation of the rat brain with a double-bridged peroxidase-antiperoxidase method. By light microscopy, the majority of varicose processes with intense TH-like immunoreactivity (LI) were contained in the hilus of the dentate gyrus (DG) and strata radiatum and lacunosum-moleculare of the CA3 region of the hippocampus. Only a few immunoreactive fibers were observed in the molecular and granule cell layers of the DG, in strata oriens and pyramidale of CA3, and in all layers of CA1. Electron microscopy confirmed that these labeled processes were primarily axons and axon terminals. Terminals with TH-LI were 0.4-1.1 micron in diameter and contained many small clear vesicles and from 0 to 3 larger dense-core vesicles. The number and types of associations formed by terminals with TH-LI were remarkably similar in the DG and hippocampus proper despite known differences in intrinsic cells and function. In both regions, the majority of terminals with TH-LI formed junctions on small (distal dendrites (52% of 112 in the DG; 67% of 116 in CA3) and dendritic spines (30% in the DG; 18% in CA3) that were both asymmetric and symmetric. In the DG, axosomatic junctions (2% of 112) were symmetric and occurred exclusively on the perikarya of granule cells, whereas junctions on large (proximal) dendrites were more numerous (16%), exhibited symmetric as well as asymmetric membrane specializations, and were of both granule (molecular layer) and nongranule (hilus) cell origin. In CA3, synaptic contacts on perikarya (5% of 116) and large (proximal) dendrites (10%) of both pyramidal cell and nonpyramidal cell origin were few and all symmetric. The distribution and types of synaptic associations formed by terminals with TH LI in the CA1 region paralleled that seen in the CA3 region. In both the dentate and hippocampus proper, 10% of the terminals with TH-LI were observed closely apposed to unlabeled terminals that formed asymmetric synapses with dendrites and dendritic spines. In rare instances, TH-immunoreactive terminals were found in close association with the basement membrane of blood vessels, astrocytic processes, or with other unlabeled terminals not forming recognizable junctions. In addition TH-LI was occasionally detected within the cytoplasm of a minority of astrocytes.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2564855 TI - Depression. PMID- 2564854 TI - Uptake of glutamic acid by Streptococcus salivarius subsp. thermophilus CNRZ 302. AB - Glutamic acid uptake in Streptococcus salivarius subsp. thermophilus was energy dependent, the source of energy and adaptation to sugar being important to efficiency of uptake. The disaccharides, lactose and sucrose, stimulated uptake, but cells grown in glucose were more active. Optimum temperature was approximately 40 degrees C and pH approximately 7.0. NaCl was strongly inhibitory to the uptake of glutamic acid although not to that of isoleucine. High specificity existed because only L-aspartic acid was inhibitory. PMID- 2564856 TI - Activation of cord T lymphocytes. I. Evidence for a defective T cell mitogenesis induced through the CD2 molecule. AB - A study was carried out on cord blood T cell activation via the CD2-mediated pathway. Despite similar percentages of circulating CD3+ and CD2+ cells in adult and cord blood, the proliferation of cord PBMC to the anti-CD3 mAb and cord T cells to anti-CD2 mAb were defective. The T cell CD3-surface structure was normally able to control CD2-mediated activation, as its modulation by a non mitogenic anti-CD3 mAb blocked cord PBMC proliferation induced by anti-CD2 mAb. CD2-stimulated cord T cells did not proliferate and did not produce a significant amount of IL-2 in culture, although they expressed the IL-2R. This observation was confirmed by the optimal proliferation of CD2-induced cord T cells when rIL-2 was added. Despite the alternative T cell activation pathway is monocyte independent in adults, the defective cord T cell activation via the CD2 molecule could also be bypassed by the addition of PMA, small amounts of either autologous or allogeneic adult and cord AC or simply rIL-1 alone. Our findings provide evidence for an intrinsic functional defect in cord CD2-mediated T cell activation, which is linked to an impaired increase of free cytoplasmic calcium, as confirmed by the effectiveness of calcium ionophore A23187 in restoring a good CD2-induced cord T cell proliferation and by measurement of cellular calcium uptake after activation via the CD2 molecule. The characteristics of cord T cells revealed by this study recall the thymocyte functional pattern and may represent functional expression of the previously described phenotypic immaturity of cord T cells. PMID- 2564857 TI - Mechanisms of immune memory. T cell activation and CD3 phosphorylation correlates with Ta1 (CDw26) expression. AB - The Ta1 (CDw26) Ag distinguishes a subset of circulating T lymphocytes that is the major population proliferating to recall Ag challenge. Unlike receptors for growth factors such as IL-2 and transferrin, the Ta1 Ag is present on T cell lines and clones irrespective of cell cycle. The appearance of Ta1 on T cells that respond to recall Ag allowed us to investigate activation requirements that may be associated with T cell immune memory. Ta1+ peripheral blood T cells were induced to proliferate by mAb recognizing either the invariant chains of the TCR, or by pairs of mitogenic antibodies directed to the CD2 molecule. In contrast, Ta1- cells were not stimulated by these antibodies. In addition, Ta1-cells did not proliferate maximally after addition of the phorbol ester PMA in combination with the calcium ionophore Ionomycin, suggesting that the intracellular targets of these agents may not be fully active. Anti-CD3-induced elevation of intracellular calcium levels was equivalent in the two subpopulations, suggesting that calcium mobilization mechanisms were intact. In contrast, PMA-induced phosphorylation of TCR CD3 chains was significantly greater in Ta1+ cells as compared to Ta1- T cells. Taken together, our results indicate that Ta1 expression, which is associated with T cell activation and memory, may be causally related to TCR and CD2-mediated activation mechanisms. The PMA inducible TCR phosphorylation in Ta1+ memory cells associated with their increased ability to proliferate after CD3/TCR or CD2 stimulation suggests that intracellular phosphorylation events may be causally associated with T cell immune memory. PMID- 2564858 TI - T cell regulation of human B cell proliferation and differentiation. Regulatory influences of CD45R+ and CD45R- T4 cell subsets. AB - The immunoregulatory functions of human T4 cell subpopulations defined by mAb to the CD45R molecule (2H4) were examined. Both CD45R- and CD45R+ T4 cells that had been treated with mitomycin C (CD45R- and CD45R+ T4-mito) provided help for the generation of Ig-secreting cells (ISC) in cultures stimulated by PWM or by immobilized mAb to CD3 (64.1). IL-2 enhanced the generation of ISC in PWM stimulated cultures and in anti-CD3-stimulated cultures containing CD45R+ T4 mito. The generation of ISC was maximal in cultures containing anti-CD3-activated CD45R- T4-mito and was not increased by IL-2. By contrast, CD45R+ T4 cells that had not been treated with mitomycin C suppressed B cell responses in cultures stimulated with PWM or anti-CD3, whereas CD45R- T4 cells suppressed the generation of ISC only in cultures stimulated with anti-CD3. IL-2 enhanced suppression by anti-CD3, but not PWM, activated CD45R- T4 cells. Suppression by CD45R+ T4 cells was maximal and not increased by IL-2. CD45R+ T4-mito were more effective suppressor-inducers in PWM-stimulated cultures, promoting the differentiation of suppressor-effector cells from CD8+ T cells. However, both CD45R+ and CD45R- T4-mito exerted comparable suppressor-inducer function in anti CD3-stimulated cultures. Moreover, in anti-CD3-stimulated cultures, T8 cells could function as both suppressor-effector cells and suppressor-inducer cells. One of the functions of suppressor-inducer cells in this system appeared to involve the production of IL-2. Thus, the addition of IL-2 facilitated the induction of suppressor-effector T8 cells by CD45R- T4-mito in PWM-stimulated cultures. Although IL-2 production by the T cell subsets varied widely depending on the nature of the stimulus, these differences could not entirely explain their capacity to function as helper cells, suppressor-effector cells or suppressor inducer cells. These results indicate that both CD45R+ and CD45R- T4 cells can help or suppress B cell responses, as well as induce suppressor-effector T8 cells. Moreover, suppressor-inducer function of T cells is not limited to the T4 cell population, but rather can also be accomplished by T8 cells. The results indicate that both T4 cell subsets and T8 cells exert multiple regulatory effects on human B cell function, with the nature of the activating stimulus playing a major role in determining the functional capacity of various T cell subsets. PMID- 2564860 TI - In vivo effects in mice of an anti-T cell immunotoxin. AB - We have evaluated both the proliferative response as well as the Thy-1 Ag expression of lymphocytes from mice treated in vivo with an anti-Thy-1 immunotoxin (IT). The IT was a rat IgG2c mAb recognizing the Thy-1 Ag, disulfide linked to a ribosome-inactivating protein isolated from the seeds of the plant Saponaria officinalis (soapwort). Toxicity studies showed that a single i.v. injection of doses up to 20 micrograms IT/mouse was well tolerated and allowed indefinite survival. The Con A-induced proliferative response of spleen cells from mice killed 1 day after treatment with sublethal doses of IT was inhibited in a dose-dependent manner, with complete inhibition observed at doses of greater than or equal to 5 micrograms IT/mouse. Control experiments showed that the inhibition was due to the IT and not to its single components. Moreover, the IT effect was abolished by a large (100-fold) excess of anti-Thy-1 mAb alone given concurrently, but not by an unrelated, isotype-matched rat mAb. At all IT doses, the proliferative response to a B cell mitogen (LPS) was normal. Kinetic studies showed a time- and dose-dependent reconstitution of Con A responsiveness. In limiting dilution cultures of spleen cells from mice treated with 5 micrograms IT 1 or 4 days before death, a 97% depletion of T lymphocytes capable of proliferation was observed. Limiting dilution cultures showed that also the thymus of IT-treated mice was depleted by more than 90% of growth-competent T lymphocytes. Cytofluorographic studies of Thy-1+ cells from the spleens of IT treated mice gave results which did not correlate with those obtained in functional assays. Thus, a dose-dependent reduction, followed by a time-dependent reconstitution of Thy-1+ cells was observed in this case too, but the depletion occurred at later time points and was less complete than that observed in functional assays. Moreover, the mean fluorescence intensity of the residual Thy 1+ cells decreased below normal levels. PMID- 2564861 TI - Specific rehabilitation for the injured recreational runner. PMID- 2564859 TI - Responsiveness of fetal and adult CD4-, CD8- thymocytes to T cell activation. AB - Day-14 fetal CD4-, CD8- thymocytes showed a greater proliferative response to PMA + IL-4 than did adult double-negative thymocytes. In contrast, adult double negative thymocytes were more responsive to PMA + IL-1 + IL-2 or to IL-1 + IL-2 alone. The adult double-negative thymocytes showed significantly greater proliferation than fetal thymocytes after stimulation via anti-CD3 or anti-Thy-1 in the presence or absence of interleukins (IL-1 + IL-2 or IL-4). Adult CD4-, CD8 thymocytes also exhibited greater calcium mobilization following anti-CD3 stimulation IL-2-dependent activation with anti-Thy-1 or IL-1 + IL-2 in the absence of PMA resulted in marked expansion of CD 3+, F23.1+, CD4-, CD8- thymocytes, a population absent in fetal thymocytes but constituting 4% of pre cultured CD4-, CD8- adult thymocytes. IL-4 + PMA failed to expand this CD 3+ population. It is hypothesized that before expression of functional TCR, T cell development may be more dependent on activation pathways not using IL-2; after TCR expression, IL-2-dependent pathways, including Thy-1-mediated stimulation, become functional. PMID- 2564862 TI - The combination of radiotherapy and chemotherapy: a review. AB - The association of radiotherapy (RT) and chemotherapy (CT) is clearly beneficial in tumours which are both chemosensitive and radiosensitive (Tubiana et al. 1987). However, contrary to the expectations, in most solid tumours this association did not lead to significant progress in long-term survival. In head and neck cancers, for example, despite a large number of controlled clinical trials, there is not yet a clear-cut demonstration of any increase in survival (DeVita et al. 1986, Tannock and Browman 1986, Taylor 1987, Tubiana et al. 1985). These disappointing results are probably due to two factors: (1) the insufficient effectiveness of the available drugs on most solid tumours; (2) the cumulative toxic effects of radiation and drugs on the normal tissues, which limits the dose of both modalities in combined treatments. PMID- 2564863 TI - Track structure analysis of ultrasoft X-rays compared to high- and low-LET radiations. AB - Monte-Carlo track structure simulations of ultrasoft X-rays, and of selected low- and high-LET radiations for comparison, have been used to obtain statistically valid frequency distributions of energy deposition in small subcellular targets which resemble the dimensions of short segments of DNA, nucleosomes and short segments of chromatin fibre. It is found that in all cases large numbers (approximately 10(3] of direct energy deposition events occur in these targets in a single mammalian cell irradiated with 1 Gy of any of these radiations. In almost all cases the numbers of energy depositions of substantial size (say, approximately greater than 100 eV in a DNA segment, approximately greater than 300 eV in a nucleosome or approximately greater than 800 eV in a segment of chromatin fibre) are also quite large, being approximately 10 to 100 per cell per Gy. It seems clear therefore that the direct effects of radiation on macromolecules must be considered in assessing the biological effects of any ionizing radiations on mammalian cells. The calculations also show that high-LET radiations can produce uniquely large energy depositions in the targets, such as are virtually unachievable by any of the other radiations; this allows the possibility of unique biochemical and cellular damage by high-LET radiations. At any realistic dose for mammalian cells, virtually all the energy depositions in these targets, from all the radiations, are due to single independent tracks; the multi-track component is negligibly small. The absolute numbers of energy depositions of approximately greater than 100 eV in DNA segments in a cell are similar to experimentally measured numbers of DNA double-strand breaks, but both these sets of numbers are one or two orders of magnitude larger than the numbers of lethal events produced in mammalian cells. The frequency of threshold energy of approximately 120 eV in a DNA segment correlates reasonably well with the relative biological effectiveness of ultrasoft X-rays and low-LET radiations for relatively radioresistant cells, but a lower threshold energy may be required for other, more sensitive, cells. PMID- 2564864 TI - Diffuse reflectance pulse radiolysis of solid DNA: the effect of hydration. AB - Using diffuse reflectance pulse radiolysis, it has been demonstrated from spectral characteristics of the resulting transients that the chemical events following irradiation of DNA depend upon its state of hydration. PMID- 2564865 TI - The role of sulphur peptide functions in free radical transfer: a pulse radiolysis study. AB - Cascading transfers of free radical centres, involving sulphur and aromatic protein functions, have been studied in further detail. The disulphide radical anion appears to be an important terminus of both oxidative and reductive radical transfer. In deaerated solutions of cysteine (20 mmol dm-3) the yield of Cys2/SS. closely resembles the yield of all primary free radicals generated by water radiolysis (.OH, H. and eaq-). The alanyl Ala/C beta., formed by electron addition to cysteine and subsequent SH- elimination, oxidizes cysteine with a rate constant of k8 = 5.0 x 10(6)dm3mol-1s-1 at pH 6 to 7 and 3.6 x 10(6)dm3mol 1s-1 at pH 9 to 10. In the case of glutathione (GSH) the eaq--induced carbon centred radical oxidizes the parent thiol with rate constants k(G. + GSH) of 7.0 x 10(6) and 1.3 x 10(6)dm3mol-1s-1 at pH 8 and pH 10, respectively; and with dithiothreitol (D(SH)2) the corresponding reaction rate is k(.DSH + D(SH)2) = 5.5 x 10(6)dm3mol-1s-1 at pH 7.0. The decarboxylated methionyl Met/C. alpha, formed by reaction of .OH with methionine, is capable of electron transfer to cystine, indicating a reduction potential for decarboxylated methione more negative than 1.6 V. The ring-closed methionyl radical cation Met/SN.+, formed by reaction of .OH with Met-Gly, oxidizes azide via equilibration, Met/SN.+ + H+ + N3- in equilibrium Met + N3., which enables an estimate to be given for the one-electron reduction potential: E degrees (Met/SN.+ + H+; Met) = +1.42 +/- 0.3 V (pH 6.8). Some further reactions of oxidizing dimeric Met2/SS.+ species in neutral solution have been demonstrated. The direction and nature of the transfers can be expressed by the scheme: (formula; see text). PMID- 2564866 TI - One-electron reduction of 5-deazalumiflavin in aqueous solution: a pulse radiolysis study. AB - The one-electron reduction of 5-deazalumiflavin has been studied in aqueous solution in the acidity range H0 = -1 to pH 13 using the reducing species CO2-, e aq and (CH3)2COH radicals. The spectral and other properties of the deazaflavin radicals formed were found to be independent of the reductant used. Four protolytic forms of the radical were distinguished with associated pKa values of 1.3 +/- 0.3, 6.0 +/- 0.3 and 10.7 +/- 0.3. PMID- 2564867 TI - Evidence for the dominance of direct excitation of DNA in the formation of strand breaks in cells following irradiation. AB - The yields of radiation-induced strand breaks measured in a plasmid DNA irradiated as a 'dry film' are similar to those measured in SV40 DNA irradiated in a cellular environment (Roots et al. 1985). This suggests a common mechanism, namely direct excitation of the DNA rather than indirectly inflicted damage from radiation-induced water radicals. This result is discussed in terms of a recently proposed mechanism of excitation transfer in DNA following direct excitation by ionizing radiation. PMID- 2564868 TI - Cell killing and DNA damage in Chinese hamster V79 cells treated with hydrogen peroxide. AB - Hydrogen peroxide induces lesions in cells similar to those from ionizing radiation, by a Fenton-type production of hydroxyl radicals. At 4 degrees C significant levels of DAN single-strand breaks (ssb) could be measured using the alkaline elution technique, after a 20-min incubation with 10(-5) mol dm-3 H2O2. Only at higher concentrations of H2O2 (greater than 10(-4) mol dm-3) where the levels of ssb measured corresponded to that induced by more than 18 Gy of X-rays, was any significant cell killing detected in a clonogenic assay. Cell killing was observed to coincide with the measurement of significant levels of DNA double strand breaks (dsb) using the filter elution technique at pH 9.6. This suggests that dsb and not ssb are important as regards hydroxyl-radical-induced cell kill, as found for ionizing radiation. The correlation of induced dsb with lethal events showed that the predicted lethal effect of the H2O2-induced dsb was approximately 5 times less than that of X-ray-induced dsb. This is compared with data previously obtained which showed differences in the lethality of dsb with the quality of radiation (Prise et al. 1987). PMID- 2564869 TI - Radiosensitization of E. coli B/r by arylhydrazonopropanedinitriles. AB - Several arylhydrazonopropanedinitriles and an arylhydrazonopropane-diethyl ester (derivatives of well-known uncouplers of oxidative phosphorylation) have been studied with respect to their ability to radiosensitize E. coli B/r under oxic and hypoxic conditions. Of the compounds studied, 2-carboxyphenylhydrazono propanedinitrile and 2-carboxyphenylhydrazonopropanediethylester were found to be the most efficient radiosensitizers under hypoxia, whilst the former compound was also found to provide radiosensitization under oxic conditions. Increased radiosensitization by 4-carboxyphenylhydrazonopropanedinitrile was observed on decreasing the pH of the irradiation incubation medium. The results are discussed with respect to the physicochemical properties of these compounds and their reactivity with thiols, for which data are presented. PMID- 2564870 TI - Biphasic survival curves for XRS radiosensitive cells: subpopulations or transient expression of repair competence? AB - Four of the most radiosensitive xrs variants of CHO-K1 cells, obtained after mutagenizing treatment with EMS, have been studied in detail over three to five decades of cell survival. Although these lines were initially reported to have very steep exponential survival curves, and to vary in sensitivity between themselves by a factor of two, we found in each case a similar biphasic response. The initial sensitivity was similar for all four lines, with a D0 of 0.5-0.7 Gy. A subpopulation, representing between 0.4 and 12 per cent of the cells, showed a resistant response, characterized by a D0 of 1.5-2.0 Gy. The previously reported variation in sensitivity seems to result from differences in the fraction of resistant cells rather than from differences in the D0. The consequence of such phenotypic variants within each cloned line is considerable, both for radiobiological studies of repair, and for molecular biology studies of the repair genes. Attempts were made to clone the sensitive and resistant subpopulations from each xrs cell line. Simple cloning from an untreated population was expected to yield pure sensitive cells, but these cells also gave biphasic responses in most cases. Only the cell line with the lowest resistant fraction (xrs5) gave a completely sensitive response in two of its subclones. Cells selected as survivors after high radiation doses were expected to yield resistant populations. However, for xrs4, 5 and 7 these subclones all gave biphasic responses. Three of the subclones from xrs6 gave biphasic responses but others gave a resistant response close to the wild type. We present a model in which transient gene expression may be seen in each individual cell if the silent copy of the xrs repair gene is temporarily hemimethylated. This transient gene transcription should occur during DNA synthesis, in the interval between synthesis of the gene and maintenance methylation. This interval may vary from cell line to cell line, resulting in different fractions of resistant cells. PMID- 2564871 TI - Expression of lethal mutations in progeny of irradiated mammalian cells. AB - The cloning efficiency of the progeny of CHO and BALB/3T3 cells surviving acute exposures of 100-1000 cGy of X-rays was examined by three different experimental protocols. A dose-dependent decrease in cloning efficiency was observed with both cell types up to 13-15 population doublings after exposure. This decrease persisted for longer times after high radiation doses; for example, the cloning efficiency of the progeny of 3T3 cells 28 population doublings after irradiation with 1000 cGy was 44 per cent of that in parallel non-irradiated controls. Confluent holding under conditions which allowed the repair of potentially lethal damage had no effect on this phenomenon. These results are consistent with the hypothesis that large numbers of lethal mutations may be expressed among the progeny of surviving cells for many generations after irradiation. PMID- 2564872 TI - Rapid repair of potentially lethal damage in normal and ataxia telangiectasia cell lines. AB - Potentially lethal damage repair (PLDR) was investigated in two normal and three ataxia telangiectasia (AT) human-skin fibroblast cell lines cultured in vitro. Using plateau-phase cells, both the time kinetics and the amount of repair were measured after irradiation. PLDR depended on both dose and survival level, as previously seen in rodent cells. Human cells differed from rodent cells in PLDR speed and the ability to discern two components within the repair response. Fast repair had a t1/2 of approximately 5-7 min while the slow response occurred over hours. AT cells had demonstrable PLDR in contrast to previous studies. Quantitatively, the proportion of fast and slow repair was similar for each dose in either normal or AT cells. However, AT cells had lower levels of both types of repair. When analyzing PLDR in human cells, differences in the rate of repair between human and rodent cells must be taken into account. PMID- 2564873 TI - Growth kinetics of C3H10T1/2 cells exposed to low-LET radiation. AB - Growth curves and size of the colonies of C3H10T1/2 cells exposed to low-LET radiation (31 MeV protons) were determined after 0, 1, 3, 5, and 7 Gy. The data show that: cell density at confluence was 3.3 x 10(4) cells/cm2; the initial division delay was very small; in the first 15 h the increase in the cell number was essentially the same at all doses; at 100 h the colony size distribution was very large, ranging from 0 to 7 generations, even within the control population. The temporal dependence of the growth properties of surviving and non-surviving cells was represented by the equation N = N0(Fe(a(t - dD] + (1 - F)ea/bD(1 - e - bD(1 - e - bD(t - dD]) where F is the surviving fraction, t the time of sampling, a the growth rate, d the division delay per unit dose, b the rate per unit of dose at which the non-surviving cells lose their ability to divide. The resulting values were: a = 0.029 +/- 0.002 h-1; b = 0.0041 +/- 0.0009 h-1 Gy-1 and d = 1 +/ 0.8 h Gy-1. It was found that growth curves are affected by non-surviving progeny up to 150, 200 and 250 h after irradiation at 3, 5 and 7 Gy, whereas at longer times the population consists essentially of progeny of surviving cells. PMID- 2564874 TI - Transport of alpha-aminoisobutyric acid into rat parotid after X-irradiation. AB - Rat parotid gland exposed to 20 Gy X-irradiation exhibits functional alteration 3 days after exposure. The flow rate of saliva and the uptake of alpha aminoisobutyric acid by the gland was reduced to 50 per cent of values for the control nonirradiated glands. When the same gland was studied in an in vitro system it functioned normally. K+ release and alpha-aminoisobutyric acid uptake by the irradiated dispersed acinar cells was comparable to the control. Transport alteration from the circulatory system into the parotid gland may cause the initial radiation-induced damage. PMID- 2564875 TI - The influence of cisplatin and unilateral nephrectomy on the response of the rat kidney to irradiation. AB - Cisplatin was administered as a single i.p. dose of 5 mg/kg to WAG/Rij rats at intervals of 7 days or 0.5 h before, or 7 days after graded X-ray doses to the left kidney. The right kidney was surgically removed 4 weeks after irradiation. Renal function was determined by measuring total urine volume excreted in 24 h, urine osmolality and serum urea. The severity of alterations in the various anatomic compartments of the kidney induced by the various treatments were graded histologically. The administration of 2 or 5 mg/kg cisplatin alone did not alter any of the kidney function parameters. Isoeffective radiation doses calculated for each of the functional parameters continuously decreased with increasing time after treatment. Differences between the isoeffective doses for the three combined treatments and for treatment with irradiation alone were only observed for urine osmolality and urine volume which primarily are tubular-related functional parameters. The histopathological grading studies also indicated that enhancement by cisplatin of radiation-induced damage was almost entirely confined to the tubules. The results of this study indicate that the sequence and length of time between treatments is an important variable in the development of cisplatin plus radiation-induced renal injury, but that none of the tested combinations showed a more than additive toxicity. PMID- 2564876 TI - Testing of methyleneiminodiacetic-catechol and other aromatic chelating agents for decorporation of 238Pu and 241Am in rats. AB - The removal of 238Pu and 241Am by five chelating agents prepared in China was compared in pilot experiments with removal by Ca-DTPA and LICAM(C). The most promising substance is quinamic acid (a methyliminodiacetic polyquinoline derivative, code name 811 or 703-73), especially in combination with Ca-DTPA. However, the best over-all reduction of both 238Pu and 241Am in all the organs studied was achieved by Ca-DTPA-administered at a ten-fold human equivalent dosage. PMID- 2564877 TI - Effect of gamma irradiation on the B vitamins of pork chops and chicken breasts. AB - A study was made of the effect of low-dose gamma irradiation on the content of thiamine (B1), riboflavin (B2), niacin, pyridoxine (B6) and cobalamin (B12) in pork chops, and thiamine, riboflavin and niacin in chicken breasts. Gamma irradiation from a caesium-137 source was used to irradiate the samples in a range of 0.49 to 6.65 kGy from -20 to +20 degrees C. Over the range of dose and temperature studied it was possible to derive a mathematical expression for predicting the losses. A calculation was made of the effect of the loss of thiamine, riboflavin and niacin due to irradiation on the overall loss of these vitamins in the American diet. The losses of riboflavin and niacin were of the order of a fraction of a per cent. Pork is an important source of thiamine, but the calculated loss at 1.0 kGy of this vitamin in cooked pork was only 1.5 per cent. There were initial increases with radiation doses up to 2-4 kGy in the measured concentrations of riboflavin and niacin in both pork and chicken. The increases were highly significant, and are of concern both to the study of radiation effects and the chemical method of the determination of these two vitamins. PMID- 2564878 TI - A single-shot rapid-mixing device for radiobiological studies with mammalian cells. AB - A single-shot rapid-mixing device is described for the rapid addition of solutions of radiation-modifying agents, to cell suspensions, at well-defined times relative to a pulse of radiation. The liquid injection system could be used to initiate or quench a wide range of chemical or biochemical reactions. The rapid-mixing device is based on a syringe driven by a stepper motor and can inject up to 2 cm3 liquid in less than 100 ms. The radiation source, a 4 MV Van de Graaff accelerator, provides an electron beam which is deflected from the beam dump on to the sample in two stages, providing a 10 ms radiation pulse. A digital delay circuit defines the interval between mixing and irradiation. The apparatus has been designed to study the kinetics of processes that occur over a time range extending from about 0.1 s to some minutes. It bridges the gap between the ranges available with conventional fast-mixing and those using standard X- or gamma irradiation methods. The time resolution of the technique has been examined by following the timecourse of radiosensitization by oxygen in mammalian cells. The timecourse of radioprotection of aerobic mammalian cells by dithiothreitol has been measured using the technique. PMID- 2564879 TI - Comments on radiation-induced G2 arrest in mouse zygotes. PMID- 2564880 TI - Benzodiazepine prescription abuse and the benzodiazepine withdrawal syndrome. AB - Benzodiazepines are among the most widely-prescribed medications in the world. Although tolerance is unlikely, abuse is widespread. Prescription abuse is especially common, with medication being taken for longer periods than intended, or at the wrong dosage, or for purposes not intended by the physician. Prolonged use, even at therapeutic levels, can produce a definite "benzodiazepine withdrawal syndrome" with disturbing symptoms such as insomnia, twitching, and restlessness, leading to prolongation of the already inappropriate usage. Suggestions for reducing this abuse are discussed. PMID- 2564881 TI - Treatment of nail puncture wounds. PMID- 2564882 TI - Two Ca current components of the receptor current in the electroreceptors of the marine catfish Plotosus. AB - In the isolated sensory epithelium of the Plotosus electroreceptor, the receptor current has been dissected into inward Ca current, ICa, and superimposed outward transient of Ca-gated K current, IK(Ca). In control saline (170 mM/liter Na), with IK(Ca) abolished by K blockers, ICa declined in two successive exponential phases with voltage-dependent time constants. Double-pulse experiments revealed that the test ICa was partially depressed by prepulses, maximally near voltage levels for the control ICa maximum, which suggests current-dependent inactivation. In low Na saline (80 mM/liter), ICa declined in a single phase with time constants similar to those of the slower phase in control saline. The test ICa was then unaffected by prepulses. The implied presence of two Ca current components, the fast and slow ICa's, were further examined. In control saline, the PSP externally recorded from the afferent nerve showed a fast peak and a slow tonic phase. The double-pulse experiments revealed that IK(Ca) and the peak PSP were similarly depressed, i.e., secondarily to inactivation of the peak current. The steady inward current, however, was unaffected by prolonged prepulses that were stepped to 0 mV, the in situ DC level. Therefore, the fast ICa seems to initiate IK(Ca) and phasic release of transmitter, which serves for phasic receptor responses. The slow ICa may provide persistent active current, which has been shown to maintain tonic receptor operation. PMID- 2564883 TI - Two syndromes of schizophrenic psychopathology associated with left vs. right temporal deficits in P300 amplitude. Four case reports. AB - In four schizophrenic patients, we examined the relationship between clinical course, including neuroleptic response, and the following biological and psychological measures: topography of the auditory P300 event-related potential, computerized tomography (CT), Andreasen's positive and negative symptom scales, the Thought Disorder Index, and a neuropsychological test battery. Two previous studies in our laboratory had shown that schizophrenic patients were differentiated from a matched normal control group by a left temporal scalp region deficit in P300 topography. This present report compares two schizophrenic patients with the typical left temporal P300 topography deficit with two schizophrenic patients with a right temporal P300 topography deficit. The two right temporal deficit patients had more positive symptoms, more thought disorder, more severely impaired functioning, earlier age of onset, poorer response to neuroleptic medications, more diffuse cognitive deficits on a neuropsychological testing battery, and poorer premorbid history than the two left temporal deficit patients. There was some evidence for the presence of more CT abnormalities suggestive of frontal lobe pathology in the right temporal deficit patients. PMID- 2564884 TI - The natural history and functional consequences of dysphagia after hemispheric stroke. AB - Data from 357 conscious stroke patients taking part in an acute intervention trial and assessed within 48 hours of the onset of symptoms, were used to investigate the prevalence and natural history of swallowing problems. Nearly 30% of patients with single-hemisphere strokes were initially found to have difficulty swallowing a mouthful of water, but in most of those who survived, the deficit had resolved by the end of the first week. Strong correlations were found between dysphagia and speech impairment (comprehension and expression) and with facial weakness, but there was no association with the side of the stroke. After controlling for other markers of overall stroke severity such as conscious level, urinary continence, white blood cell count and strength in the affected limbs, swallowing impairment still showed a significant inverse correlation with functional ability at 1 and 6 months. These results indicate that, even if dysphagia itself is not responsible for much excess mortality in acute stroke, it might still lead to complications which hamper functional recovery. PMID- 2564885 TI - Excitatory amino acid-stimulated uptake of 22Na+ in primary astrocyte cultures. AB - In this study we have found that L-glutamic acid, as well as being taken up by a Na+-dependent mechanism, will stimulate the uptake of 22Na+ by primary astrocyte cultures from rat brain in the presence of ouabain. By simultaneously measuring the uptake of 22Na+ and L-3H-glutamate a stoichiometry of 2-3 Na+ per glutamate was measured, implying electrogenic uptake. Increasing the medium K+ concentration to depolarize the cells inhibited L-3H-glutamate uptake, while calculations of the energetics of the observed L-3H-glutamate accumulation also supported an electrogenic mechanism of at least 2 Na+:1 glutamate. In contrast, kinetic analysis of the Na+ dependence of L-3H-glutamate uptake indicated a stoichiometry of Na+ to glutamate of 1:1, but further analysis showed that the stoichiometry cannot be resolved by purely kinetic studies. Studies with glutamate analogs, however, showed that kainic acid was a very effective stimulant of 22Na+ uptake, but 3H-kainic acid showed no Na+ -dependent uptake. Furthermore, while L-3H-glutamate uptake was very sensitive to lowered temperatures, glutamate-stimulated 22Na+ uptake was relatively insensitive. These results indicate that glutamate-stimulated uptake of 22Na+ in primary astrocytes cultures cannot be explained solely by cotransport of Na+ with glutamate, and they suggest that direct kainic acid-type receptor induced stimulation of Na+ uptake also occurs. Since both receptor and uptake effects involve transport of Na+, accurate measurements of the Na+ :glutamate stoichiometry for uptake can only be done using completely specific inhibitors of these 2 systems. PMID- 2564886 TI - Roles for mitotic history in the generation and degeneration of hippocampal neuroarchitecture. AB - The mechanisms regulating the highly ordered neuroarchitecture of the mammalian brain are largely unknown. The present study took advantage of hippocampal pyramidal-like neurons that arose from a common progenitor cell in cell culture (sister neurons) to ascertain the contribution of intrinsic factors to both the generation and degeneration of neuroarchitecture. Sister neurons were similar in overall cell form and dendritic numbers and lengths. Control non-sister neurons that grew in contact did not generate similar morphologies, indicating that the similarity of sister cells did not result from influences of the local microenvironment or cell interactions. These results suggest that intrinsic factors related to mitotic history play a role in the generation of neuroarchitecture. Since particular groups of hippocampal neurons are sensitive to glutamate neurotoxicity in situ and are vulnerable in neurodegenerative disorders, it was of interest to test glutamate sensitivity in the neuronal population and in mitotic sister neurons. A subpopulation of pyramidal neurons was sensitive to glutamate neurotoxicity. A striking finding was that sister neurons were invariably either both sensitive or both resistant to glutamate, while non-sister neurons often showed different responses to glutamate. Pharmacological studies indicated that glutamate neurotoxicity was mediated by kainate/quisqualate type receptors by a mechanism involving calcium influx through membrane channels. Fura-2 measurements of intracellular calcium revealed that sister neurons had similar rest levels of calcium and, strikingly, glutamate caused a dramatic increase in intracellular calcium levels only in neurons which subsequently degenerated. Apparently, intrinsic differences in sensitivity to glutamate lie at a point prior to calcium entry, probably at the level of glutamate receptors. Taken together, these results indicate that the mitotic history of a neuron can determine its presence and potential for connectivity as well as its susceptibility to neurodegeneration. PMID- 2564887 TI - Summary of the NATO advanced research workshop on dietary omega 3 and omega 6 fatty acids: biological effects and nutritional essentiality. AB - A number of human studies presented at the workshop indicate that the premature infant at birth is biochemically deficient in docosahexaenoic acid (DHA) in both the brain and liver phospholipids, and that DHA is essential for normal visual acuity. The amount of DHA necessary to maintain normal amounts of the liver and brain phospholipids postnatally is 11 mg/kg daily. Elderly patients on long-term gastric tube feedings and others on long-term intravenous fluids and on total parenteral nutrition are particularly prone to deficiencies of alpha-linolenic acid, eicosapentaenoic acid (EPA) and DHA. The amounts estimated to prevent deficiencies in the elderly are 800-1100 mg/d of alpha-linolenic acid and 300-400 mg/d of EPA and DHA combined. Preliminary data indicate that children with malnutrition and mucoviscidosis, women with toxemia, and elderly people have decreased amounts of DHA in plasma phospholipids. The omega 3 fatty acids lower triglycerides and, at high levels, lower cholesterol. The anti-aggregatory, anti thrombotic and anti-inflammatory properties of omega 3 fatty acids have been confirmed, and a dose-response curve is emerging. Despite the increase in bleeding time, no clinical evidence of bleeding has been noted by the investigators in any of the studies. Clinical trials are necessary in order to precisely define the dose and mechanisms involved in defining the essentiality of omega 3 fatty acids in growth and development and their beneficial effects in coronary heart disease, hypertension, inflammation, arthritis, psoriasis, other autoimmune disorders, and cancer. PMID- 2564888 TI - Genetic evidence for the role of Thy-1 in neurite outgrowth in the mouse. AB - Non-neuronal accessory cells of mouse dorsal root ganglion cultures secrete a complex that stimulates neurite outgrowth in neonatal sympathetic ganglion neurons. A monoclonal antibody that binds to these two cell types also immunoprecipitates neurite outgrowth complex from mouse conditioned medium and binds to mouse brain tissue. Genetic analysis of the component recognized by the antibody revealed that it maps to the Thy-1 locus on mouse chromosome 9. Further studies of cell-type specificity, sedimentation analysis and antibody competition, confirmed that it is indistinguishable from the product of the Thy-1 locus. The finding of an association between Thy-1 and neurite outgrowth complex in the mouse argues for a role of the Thy-1 locus in the interactions of neurons with their surroundings. PMID- 2564889 TI - Evidence for allosteric blockade of serotonergic receptors in rabbit thoracic aorta. AB - 2-Brom-d-lysergic acid diethylamide (BOL) appeared to behave as either a reversible competitive or noncompetitive antagonist of vascular serotonergic (5 HT2) receptors depending on experimental conditions. This was explored using rabbit thoracic aorta rings mounted in tissue baths for the measurement of isometric contraction. BOL caused concentration-dependent parallel rightward shifts of the 5-HT dose-response curve in untreated aortas but, in addition, caused a marked reduction of maximal response in aortas pretreated with benextramine to inactivate alpha adrenoceptors. Ketanserin behaved as a reversible competitive antagonist in both untreated and benextramine-pretreated aortas. The respective ketanserin pA2 values were 9.08 +/- 0.09 (S.E.M.) and 9.01 +/- 0.04. Ketanserin, 1 x 10(-7) M, reversed completely the reduction of maximal response caused by 1 x 10(-9) M BOL and partially reversed those caused by 1 x 10(-8) and 1 x 10(-7) M BOL. Furthermore, the 5-HT dose-response curve in the presence of 1 x 10(-7) M ketanserin and that in the presence of 1 x 10(-7) M ketanserin plus 1 x 10(-9) M BOL were superimposed. These results are consistent with the conclusion that BOL is a noncompetitive 5-HT2 receptor antagonist. We propose that BOL acts at an allosteric site to convert the 5-HT2 receptor to a low activity state, thus, reducing the maximal response. BOL does not act at the 5-HT2 receptor itself. Ketanserin competes with 5-HT at the 5-HT2 receptor and with BOL at the allosteric site.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2564890 TI - Differential effects of dopaminergic drugs on open-field behavior of spontaneously hypertensive rats and normotensive Wistar-Kyoto rats. AB - Brain catecholamines may play a role in the development of hypertension in the spontaneously hypertensive rat (SHR). To investigate central dopaminergic function in this strain, the effect of various dopaminergic drugs on open-field activity was assessed. In general, basal levels of ambulation were similar in SHRs and their controls, the Wistar-Kyoto rats (WKY). Exploratory rearing activity was increased in SHRs, however. Haloperidol inhibited open-field ambulation in SHRs and WKY, but higher doses were needed in the SHR. Apomorphine inhibited ambulatory activity in WKY but had virtually no effect in SHRs. Amphetamine and the dopamine uptake inhibitor GBR-12909 increased ambulation in SHRs and WKY to a similar extent. Central administration of haloperidol or sulpiride decreased ambulatory activity in WKY, but had no effect in SHRs at the doses used. SCH-23390 decreased ambulation scores both in SHRs and WKY, with the effect being slightly greater in the latter strain. In all cases exploratory rearing was affected as well by the drugs used. The large difference in base-line scores has to be taken into account when interpreting these data, however. Also, for comparison the noradrenergic drugs clonidine and desipramine were tested. Both elicited decreases in activity which were identical in SHRs and WKY. These results show that a number of dopaminergic drugs have differential effects on open-field activity of SHRs as compared to WKY. Ambulatory activity appeared more resistant to inhibition in SHRs than in WKY. The possibility that these results could reflect altered dopaminergic function in SHR involved in the development of hypertension in this strain is discussed. PMID- 2564891 TI - Interaction acetylcholine-glutamate in rat hippocampus: involvement of two subtypes of M-2 muscarinic receptors. AB - The effects of acetylcholine (ACh) on the release of endogenous glutamic acid (GLU) and of [3H]ACh have been investigated comparatively in superfused rat hippocampal synaptosomes. Exogenous ACh added to the superfusion fluid inhibited the Ca++-dependent K+ (15 mM)-evoked release of GLU in a concentration-dependent manner (the maximal inhibition was about 50%). Carbachol and oxotremorine mimicked, although less potently, the action of ACh. The inhibition of GLU release caused by 10 microM ACh was antagonized by 0.1 microM atropine but not by 10 microM mecamylamine. It also was insensitive to the M-1 receptor antagonists pirenzepine or dicyclomine (both at 1 microM). In contrast, the novel M-2 muscarinic antagonist AF-DX 116 [(11-[(2-[diethylamino]methyl)-1 piperidinyl]acetyl)-5-11-dihydro-6 H-pyrido-[2-3-b][1,4]benzo-diazepine-6-one] was as potent as atropine in blocking the inhibition of GLU release brought about by ACh. The autoreceptor-mediated inhibition of [3H]ACh release observed in presence of ACh (10 microM) was totally antagonized by atropine (0.1 microM). It was insensitive to mecamylamine (10 microM), dicyclomine (1 microM) or pirenzepine (1 microM). However, it was much less sensitive to AF-DX 116 (80-100 times) than the cholinergic inhibition of GLU release. It is concluded that 1) the release of GLU in rat hippocampus can be inhibited through a muscarinic receptor and 2) this novel muscarinic receptor belongs to the M-2 subtype but it seems to differ pharmacologically from the M-2 autoreceptor. PMID- 2564892 TI - Structure-activity study on the actions of taxol and related taxanes on primary cultures of adrenal medullary cells. AB - Taxol is a natural plant product which has antimitotic activity. It is currently being investigated for use both as a cancer chemotherapy agent and as a tool to investigate microtubule involvement with various cellular functions. In the studies reported here, the actions of taxol on cultured adrenal medullary cells are compared to those of two other taxanes, baccatin III and 19-hydroxybaccatin III, which are structurally similar but lacking a bulky ester linkage at the C13 position. Unlike taxol, baccatin III and 19-hydroxybaccatin III have little or no known actions on mammalian microtubules. These studies were designed 1) to establish structural requirements for taxol's effects on adrenal cells and 2) to investigate whether the effects of taxol on adrenal cells involve microtubules. The effects of taxol on basal catecholamine release, on nicotinic receptor stimulated catecholamine release and on the adrenal microtubule network are quantitatively and qualitatively different from those of baccatin III and 19 hydroxybaccatin III. These results suggest that esterification of taxol at position C13 is important in taxol's inhibitory actions on nicotinic receptor stimulated catecholamine release. Similar structure requirements have been reported for taxol's actions on microtubules. Catecholamine release stimulated through activation of nicotinic receptors is much more sensitive to the inhibitory actions of taxol than is release stimulated by other secretagogues. Finally, a direct effect of taxol on the adrenal microtubular network was demonstrated using treatment conditions which are similar to those used to interfere with nicotinic receptor function. The studies reported here offer further evidence that taxol's actions on nicotinic receptor function may involve microtubules. PMID- 2564893 TI - Pharmacological characterization of the receptor mediating electrophysiological responses to dopamine in the rat medial prefrontal cortex: a microiontophoretic study. AB - The pharmacological profile of the receptor mediating inhibitory effects of dopamine (DA) in the rat medial prefrontal cortex (PFC) was characterized using extracellular single unit recording and microiontophoretic techniques. Iontophoretic application of DA inhibited 65% of spontaneously active cells in the deep layers of the PFC, while producing little effect on cells in superficial laminae. The D2 selective antagonist, sulpiride, specifically attenuated DA induced inhibition of deep layer PFC neurons, without blocking the inhibitory effects of gamma-aminobutyric acid (GABA) or serotonin (5-HT). Surprisingly, sulpiride antagonism did not appear to be stereospecific, as both its (-)- and (+)-isomers proved equally effective at blocking the inhibitory effects of DA. In contrast to sulpiride, the D1 selective antagonist, SCH23390, was much less effective at attenuating inhibitory responses to DA. The effects of selective agonists also were examined on DA-sensitive PFC neurons. The D2 selective agonist, LY171555, and the D1 selective agonist, SKF38393, produced inhibitory effects on a small number of DA-sensitive PFC neurons. However, the majority of cells tested were inhibited only by DA and not by LY171555 or SKF38393. In addition, coiontophoresis of LY171555 and SKF38393. In addition, coiontophoresis of LY171555 and SKF38393 failed to inhibit the majority of DA-sensitive PFC cells tested. LY171555, but not SKF38393, significantly attenuated DA-induced inhibition when applied simultaneously, suggesting that the D2 selective agonist might possess partial agonist/weak antagonist activity at this receptor. These results indicate that the receptor mediating the inhibitory effects of DA in the medial PFC has the pharmacological characteristics of a D2 subtype. However, this receptor may not be identical to D2 sites in other brain regions. PMID- 2564894 TI - Synthesis of congeners and prodrugs. 3. Water-soluble prodrugs of taxol with potent antitumor activity. AB - Taxol has shown good in vivo antitumor activity in a number of test systems. The formulation of taxol for antitumor testing has been difficult. Esterification at either C-2' or C-7 resulted in loss of in vitro tubulin assembly activity but not cytotoxicity. These observations suggested that esters at C-2' and/or C-7, which would tend to promote water solubility, might serve as useful prodrugs of taxol. The reaction of taxol with either succinic anhydride or glutaric anhydride in pyridine solution at room temperature gave the crystalline mono 2'-adducts 1b and 1f, respectively. Salts of these acids (1b, 1f, 1i) were formed by the addition of 1 equiv of the corresponding base, followed by evaporation and/or freeze drying of the solvent(s). The salts had improved antitumor activity as compared to the free acids. The triethanolamine and N-methylglucamine salts showed greatly improved aqueous solubility and were more active than the sodium salts. The glutarate series was preferred because of the higher activity and the higher yields obtained. 2'-Glutaryltaxol (1f) was coupled with 3-(dimethylamino)-1 propylamine, using CDI, to form in excellent yield the amino amide 1o. The hydrochloride salt (1p) showed good solubility and was extremely potent and active. At 10 mg/kg, in the B16 screen, 1p gave a T/C of 352 with 5 out of 10 cures. In the MX-1 breast xenograft assay, this prodrug gave values of -100 at doses of 40 and 20 mg/kg, with all live animals being tumor free. PMID- 2564896 TI - Role of the first assistant in ophthalmic surgery. AB - 1. As the number of Ambulatory Surgery Centers increases and the Medicare reimbursements for surgeon first assistants decline, the need for non-physician first assistants grows. 2. Qualifications for a first assistant include knowledge skill, and devotion to this speciality. 3. The first assistant is part of a medical team that consists of the surgeon, the first assistant, the scrub nurse or technician, and the circulating nurse. 4. The role of the first assistant helps to increase efficiency in the operating room and enhance the quality of patient care. PMID- 2564895 TI - Identification of structural requirements for analogues of atrial natriuretic peptide (ANP) to discriminate between ANP receptor subtypes. AB - The structure-activity relationships for affinity and selective binding of atrial natriuretic peptide (ANP) and analogues to guanylate cyclase coupled (CC) and non cyclase coupled (NC) receptors in rabbit lung membranes are described. We have designed a series of peptides to try to identify the minimal sequence involved in specific recognition of each receptor subtype. The affinity of the peptides was determined from competitive binding experiments. Several peptides derived from the rat ANP sequence, e.g., des-[Phe106, Gly107, Ala115, Gln116]ANP-(103-125)NH2 (4), des-[Cys105,121]ANP-(104-126) (5), and [Acm-Cys105]ANP-(105-114)NH2 (9) have high affinity and selectivity for the noncoupled site. Peptide 4 was the most selective ligand with an affinity superior to that of ANP-(103-126). This compound does not displace the radiolabeled ligand from the guanylate cyclase coupled receptor at the highest concentration tested (100 nM). The structure activity relationship for affinity and selectivity is discussed. Comparison of the peptide sequences suggests that the structural feature responsible for recognition of the NC site resides in a single sequence of seven contiguous amino acids from the cyclic core of the hormone. The corresponding heptapeptide retains affinity to the guanylate cyclase uncoupled binding site and is proposed to encompass the minimal sequence for specific recognition of the non-guanylate cyclase coupled ANP receptor. PMID- 2564897 TI - Training program for new ophthalmic assistants. AB - 1. A certified technician supervises the training of new ophthalmic assistants, but the entire technical staff is involved in the instruction. 2. The program lasts for 10 weeks: 8 weeks of training with ophthalmic procedures and 2 weeks of observation. 3. The essential elements of a well-rounded training program include knowledgeable trainers, role playing, and hand-on experience. PMID- 2564898 TI - Evidence for a role of virulent human immunodeficiency virus (HIV) variants in the pathogenesis of acquired immunodeficiency syndrome: studies on sequential HIV isolates. AB - Sequential human immunodeficiency virus (HIV) isolates, recovered from a panel of longitudinally collected peripheral blood mononuclear cells obtained from 20 initially asymptomatic HIV-seropositive homosexual men, were studied for differences in replication rate, syncytium-inducing capacity, and host range. Eleven individuals remained asymptomatic; nine progressed to acquired immunodeficiency syndrome (AIDS) or AIDS-related complex (ARC) at the time point at which the last HIV isolate was obtained. In 16 individuals, only non-syncytium inducing (NSI) isolates, with a host range restricted to mononuclear cells, were observed. From four individuals, high-replicating, syncytium-inducing (SI) isolates that could be transmitted to the H9, RC2A, and U937 cell lines were recovered. From two of these four individuals, SI isolates were obtained throughout the observation period. In the two others, a transition from NSI to SI HIV isolates was observed during the period of study. Three of these four individuals developed ARC or AIDS 9 to 15 months after the first isolation of an SI isolate. With the exception of the two individuals in whom a transition from NSI to SI isolates was observed, within a given individual the replication rate of sequential HIV isolates was constant. A significant correlation was found between the mean replication rate of isolates obtained from an individual and the rate of CD4+ cell decrease observed in this individual. In individuals with low replicating HIV isolates, no significant CD4+ cell loss was observed. In contrast, recovery of high-replicating isolates, in particular when these were SI isolates, was associated with rapid decline of CD4+ cell numbers and development of ARC or AIDS. These findings indicate that variability in the biological properties of HIV isolates is one of the factors influencing the course of HIV infection. PMID- 2564900 TI - [Sport and drugs--when is there a risk for doping? All beta blockaders and diuretics are now prohibited]. PMID- 2564899 TI - Carrier identification of cystic fibrosis by recombinant DNA techniques. AB - As a result of recent advances in molecular genetics, carrier testing for cystic fibrosis (CF) is now available as a clinical assay in a limited number of laboratories. Because neither the gene nor the mutation for this disorder has yet been defined, the analysis relies on an indirect approach that uses DNA sequence polymorphisms and linkage analysis. With use of this general approach, several family members, in addition to those persons seeking carrier information, must be tested. Currently, more than 97% of families are "fully informative" when these markers are used in a linkage analysis; thus, the carrier status of most persons who have a relative with CF can be determined. Recently, strong linkage disequilibrium has been shown between two polymorphic loci (defined by the DNA probes KM.19 and XV-2c) and the CF locus. Because of this important finding, haplotype testing can be used in many clinical settings, such as for families in which a DNA sample is not available from the affected person or for those families in which one spouse has no family history of CF and the other is either affected or is at a high risk of carrying the CF mutation. Overall, the application of recombinant DNA techniques has greatly enhanced the ability to determine, with a high level of accuracy, the carrier status of those persons at risk for inheriting the CF mutation. PMID- 2564901 TI - Natural history of localised prostatic cancer. A population-based study in 223 untreated patients. AB - In a population-based study, disease progression and survival were evaluated in untreated patients with newly diagnosed cancer of the prostate without distant metastases. Complete follow-up was achieved in 223 of 227 (98%) consecutively diagnosed, eligible patients of all ages. After 5 years, the cumulative progression-free survival (with 95% confidence interval) was 71.8 (65.5-78.1)% and survival corrected for causes of death other than prostatic cancer was 93.8 (88.3-97.6)%. Univariate and multivariate analyses showed no association between age at diagnosis and the natural course. Local progression was less common in localised, non-palpable tumours than in larger tumours. The rate of progression was 18.7 (6.1-57.1) times higher and that of disease-specific death 216.0 (31.2 1496) times higher in patients with poorly than in those with highly differentiated tumours. It is concluded that tumour grade at diagnosis is an excellent predictor of local and distant progression. The low death rate, especially in patients with highly and moderately differentiated tumours, means that any local or systemic therapy intended for patients with early prostatic cancer must be evaluated in clinical trials with untreated controls for comparison. PMID- 2564902 TI - Chronic relapsing pericarditis and dilated cardiomyopathy: serological evidence of persistent enterovirus infection. AB - By means of two different IgM-capture assays, enterovirus-specific IgM responses were shown in 9 of 14 (64%) patients with chronic relapsing pericarditis. This finding suggests persistent enterovirus infection, particularly coxsackie B virus infection. IgM responses persisted for at least 1 year and for up to 10 years after onset of symptoms. In contrast, patients with acute enterovirus infections, including acute pericarditis, had transient responses. Among patients with acute pericarditis, the level of IgM antibody was significantly higher in those who subsequently relapsed (mean 1.21, range 0.6-2.0 optical density [OD] units) than in those who did not (0.4, 0.2-0.9 OD units; p less than 0.01). Of 86 patients with dilated cardiomyopathy, 28 (33%) showed enterovirus-specific IgM responses which were present for up to 19 months before transplantation and persisted up to 4 years afterwards. Although the distribution of HLA types in these patients was similar to that in the general population, the frequency of the HLA A2 haplotype was significantly higher in those who were IgM positive. IgM antibody was significantly more common in those who had had symptoms for longer than a year before transplantation than in those with a shorter duration of symptoms (1 of 21 vs 8 of 23; p less than 0.02). Persistent virus-specific serum IgA responses were also shown in patients with chronic cardiac disease. PMID- 2564903 TI - Intense microvascular constriction after angioplasty of acute thrombotic coronary arterial lesions. AB - Immediately after balloon dilation of a fresh thrombotic coronary lesion, 5 patients had angina, ST segment elevation, and a striking reduction of blood flow in the dilated artery. A mean (SEM) pressure gradient across the dilated lesion of only 3(1) mm Hg and an average minimum lesion diameter of 1.7 mm indicated that the decline in resting blood flow was not due to obstruction at the site of the original lesion. Neither distal vascular emboli nor side branch occlusions were visible on the angiogram. An increase in distal coronary artery pressure during a subsequent balloon inflation suggested that the site of vasoconstriction was distal to the origin of collateral vessels. The syndrome lasted 48-80 min and was not reversed with nitroglycerin or thrombolytic drugs. Papaverine lessened the syndrome transiently on one occasion. Such microvascular constriction, caused by release of potent vasoconstrictors from the clot, may partly explain the failure of emergency angioplasty to reduce infarct size in acute myocardial infarction. PMID- 2564904 TI - Investigations in children who were in utero at onset of insulin-dependent diabetes in their mothers. AB - 55 children who were in utero when type 1 diabetes developed in their mothers were studied at a mean (SEM) age of 10.4 (0.6) years: only 1 was diabetic. Biochemical and immunological indices, measured in 35 children, showed no evidence of beta cell dysfunction. Thus, the fetal beta cells seem to be unaffected by the mechanisms that cause diabetes in their mothers. PMID- 2564905 TI - Health in the greenhouse. PMID- 2564906 TI - Alternatives to growth hormone. PMID- 2564907 TI - Homing in on Wilson's disease. PMID- 2564908 TI - Retinal vasculitis. PMID- 2564909 TI - Blind injustice. PMID- 2564910 TI - Haemopoietic growth factors 1. PMID- 2564912 TI - Patient-applied podofilox for treatment of genital warts. AB - In a double-blind trial, 0.5% podofilox (podophyllotoxin) or placebo was applied by patients to their own genital warts in up to four treatment cycles. At some time during the study, 25 of the 56 podofilox treated patients and none of the 53 placebo group were completely wart-free. At the end of the treatment, 73.6% of the original warts in podofilox treated patients were gone compared with only 8.3% of those in the placebo group (mean percentage of total original wart area was reduced by 82.3% compared with 4.2%). 82% of the treated warts in the podofilox group and 13% in the placebo group had resolved at 6 weeks. Recurrence was observed in 34% of the previously resolved warts. Consistent with this rate of recurrence, new warts developed in a third of the subjects in each group at sites remote from the treatment site. There were no systemic adverse reactions, although transient inflammation, erosion, pain, and burning were common. PMID- 2564911 TI - Prevalence of and mortality from human immunodeficiency virus type 2 in Bissau, West Africa. AB - In a community based prevalence study of HIV infection in Bissau, West Africa, 1987, the population in 100 randomly selected "houses" was asked to participate. 89% (1329/1499) were examined and had a blood sample taken. None was HIV-1 seropositive but 4.7% were seropositive for HIV-2 (0.6% in children, 8.9% in those aged 15 years and over, and 20% in those aged 40 years and over). There was no significant difference in seroprevalence between areas or ethnic groups or between individuals of different civil status when age was taken into account. Sexual contact and blood transfusions were the dominant transmission routes, and no case of vertical transmission was identified. The HIV-2 seroprevalence in spouses of HIV-2 seropositive index persons was 40%. For a history of blood transfusion the relative risk of being HIV-2 seropositive was 103.6 in children and 2.4 for adults. After exclusion of spouses, no clustering of HIV-2 seropositivity was seen. At follow-up, after a mean observation time of 325 days, there was an excess mortality for HIV-2 seropositives. The relative risk of dying for HIV-2 seropositive children was 60.8 and for adults 5.0. PMID- 2564913 TI - Adrenaline response to hypoglycaemia and insulin species. PMID- 2564914 TI - Duodenal ulcer relapse after eradication of Campylobacter pylori. PMID- 2564915 TI - Trends in lymphoma diagnosis. PMID- 2564916 TI - Routine coronary angiography for effort angina. PMID- 2564918 TI - Embryo biopsy. PMID- 2564917 TI - Cardiac failure responding to growth hormone. PMID- 2564919 TI - Fetal brain tissue. PMID- 2564920 TI - 99mTc-HMPAO washout in prognosis of stroke. PMID- 2564921 TI - Caffeinated beverages and decreased fertility. PMID- 2564923 TI - Intrauterine ultrasonography and endometrial cancer. PMID- 2564924 TI - Umbilical cord care and neonatal tetanus. PMID- 2564922 TI - Hepatitis B in mental handicap hospitals. PMID- 2564925 TI - Vaginal ultrasound. PMID- 2564926 TI - Detection of bone marrow lesions. PMID- 2564927 TI - Disproportionate antecollis in multiple system atrophy. PMID- 2564929 TI - Duration of contraception after etretinate. PMID- 2564928 TI - Prevention of NSAID-induced gastric ulcer with prostaglandin analogues. PMID- 2564930 TI - Cardiovascular and renal effects of ramipril. PMID- 2564931 TI - Lisinopril and renal failure. PMID- 2564932 TI - Abdominal incision. PMID- 2564933 TI - Innocent elevation of aspartate aminotransferase. PMID- 2564934 TI - Ciguatera and military activity. PMID- 2564935 TI - Food irradiation. PMID- 2564936 TI - Complaints and litigation. PMID- 2564937 TI - Eleventh week amniocentesis for prenatal diagnosis of metabolic diseases. PMID- 2564938 TI - Third World debts. PMID- 2564939 TI - Transoesophageal ultrasound imaging of spinal canal and cord. PMID- 2564940 TI - Corticosteroid induced osteoporosis and hormone implants. PMID- 2564941 TI - Food-borne listeriosis. PMID- 2564942 TI - Possible role of person-to-person transmission of Listeria infection in immunocompromised patients. PMID- 2564943 TI - Bartonellosis and Kaposi sarcoma of AIDS. PMID- 2564944 TI - Munchausen AIDS. PMID- 2564945 TI - Replicative and cytopathic characteristics of HIV-2 and severity of infection. PMID- 2564946 TI - Munchausen Zollinger-Ellison syndrome. PMID- 2564947 TI - Legal power to sterilise incompetent women. PMID- 2564948 TI - Trichophyton asthma: sensitisation of bronchi and upper airways to dermatophyte antigen. AB - 12 adult patients with perennial asthma and chronic skin infection were found to have immediate hypersensitivity to Trichophyton spp. 10 patients were tested by bronchial provocation and gave immediate bronchial reactions to an extract of T tonsurans. Double-blind, placebo-controlled nasal challenge of 8 patients demonstrated that the upper airways of these patients were also sensitive to this dermatophyte antigen. In addition to perennial asthma most of the patients had persistent eosinophilia and chronic rhinosinusitis. The results suggest that absorption of fungal antigen can give rise to IgE antibody production, sensitisation of the airways, and symptomatic asthma and rhinosinusitis. Several patients had many of the features of late onset or "intrinsic" asthma. PMID- 2564949 TI - Randomised double-blind trial of recombinant pro-urokinase against streptokinase in acute myocardial infarction. PRIMI Trial Study Group. AB - In a prospective, double-blind, randomised trial, 401 patients with a first acute myocardial infarction were treated within 4 h of onset of symptoms with 80 mg recombinant pro-urokinase or single-chain urokinase plasminogen activator (rscu PA; proposed INN saruplase) intravenously given as a 20 mg bolus followed by 60 mg infusion for 60 min (198 patients), or 1.5 million IU streptokinase infused over 60 min (203 patients). The first two angiograms were taken at 60 min and at 90 min. Angiography was repeated at 24-36 h. Patency rates at 60 min were 71.8% for rscu-PA and 48.0% for streptokinase (p less than 0.001) and at 90 min they were 71.2% and 63.9%, respectively (p = 0.15). At 24-36 h 6/121 patients treated with rscu-PA and 5/114 patients treated with streptokinase showed reocclusion of the vessel. At the end of the thrombolytic infusion (60 min) fibrinogen concentration had decreased to 0.44 (0.23-1.27) g/l (median, 1st and 3rd quartile) in patients treated with rscu-PA and to 0.17 (0.06-0.27) g/l in patients treated with streptokinase (p less than 0.001). Concentrations of fibrin(ogen) degradation products rose to 96 (24-240) mg/l after rscu-PA and to 240 (192-360) mg/l after streptokinase (p less than 0.001). Bleeding complications were less common in the rscu-PA than in the streptokinase group (p less than 0.01). Thus intravenous rscu-PA led to higher patency rate, earlier reperfusion, less disturbance of haemostasis, and fewer bleeding complications than did intravenous administration of streptokinase. PMID- 2564950 TI - Cholesterol as risk factor for mortality in elderly women. AB - 92 women aged 60 years and over (mean 82.2, SD 8.6) living in a nursing home and free from overt cancer were followed-up for 5 years. 53 died during this period; necropsy revealed cancer in only 1 patient. Serum total cholesterol at entry ranged from 4.0 to 8.8 mmol/l (mean 6.3, SD 1.1). Cox's proportional hazards analysis showed a J-shaped relation between serum cholesterol and mortality. Mortality was lowest at serum cholesterol 7.0 mmol/l, 5.2 times higher than the minimum at serum cholesterol 4.0 mmol/l, and only 1.8 times higher when cholesterol concentration was 8.8 mmol/l. This relation held true irrespective of age, even when blood pressure, body weight, history of myocardial infarction, creatinine clearance, and plasma proteins were taken into account. The relation between low cholesterol values and increased mortality was independent of the incidence of cancer. PMID- 2564951 TI - Prevalence of diabetes and impaired glucose tolerance in rural Tanzania. AB - The prevalence of diabetes mellitus and impaired glucose tolerance (IGT) was assessed by use of WHO diagnostic criteria in 6299 Africans aged 15 years and above living in six villages in Tanzania. 0.87% (1.1% male, 0.68% female) had diabetes and 7.8% (6.9% male, 7.7% female) had IGT. Prevalence rates were 1.1% and 8.4%, respectively, when age-adjusted to the USA population. Only 7 (13.5%) of the 53 individuals with diabetes had been known to have the disorder; 34 (74%) of the other 46 were symptom-free. Mean age was 54 (SD 20) for diabetic subjects and 37 (17) years for the whole population. Diabetes and IGT rates did not differ significantly between villages despite geographical, socioeconomic, and dietary differences. Diabetes rates increased modestly with age and body mass index (BMI). Fasting blood glucose (FBG) levels did not rise significantly with age but correlated positively with systolic blood pressure (BP) and negatively with haemoglobin concentration (Hb) and BMI. The 2 hour post-glucose load blood glucose values correlated positively with age, sex, and systolic BP and negatively with Hb. Diabetes is less prevalent in rural Africa than in developed countries, even when age has been corrected for. This difference is probably related to body weight, diet, and exercise. PMID- 2564952 TI - Novel lysosomal glycoaminoacid storage disease with angiokeratoma corporis diffusum. AB - A 46-year-old Japanese woman had disseminated angiokeratoma, confirmed by electron microscopy which showed numerous cytoplasmic vacuoles in cells of the kidney and skin. Enzyme activities against synthetic and natural substrates in leucocytes and fibroblasts were normal. Her urine contained a large amount of sialylglycoaminoacids, with predominant excretion of an O-glycoside-linked glycoaminoacid. PMID- 2564953 TI - Abortion USA. PMID- 2564954 TI - Signals, transducers, and secretion. PMID- 2564955 TI - Brain damage by neonatal hypoglycaemia. PMID- 2564956 TI - Visual evoked potentials and nonfunctioning pituitary tumours. PMID- 2564957 TI - Xerostomia. PMID- 2564958 TI - Haemopoietic growth factors 2: clinical applications. PMID- 2564959 TI - Mammography in symptomatic breast disease. AB - The contribution of mammography to the diagnosis of breast cancer was examined in 5080 patients with various breast symptoms. There were 562 breast cancers within this group. The sensitivity of mammography was 88%. Only 18 cancers were detected by mammography alone, and of these 3 were in the other breast. 7 of the remaining 15 cancers had clinical signs which would have been pursued to open biopsy in the absence of mammography. Clinical examination is of paramount importance in the management of patients with symptomatic breast disease. Mammography is most useful when applied in specific situations rather than to screen every patient with breast symptoms. PMID- 2564961 TI - Aspergillus, asthma, and amoebae. PMID- 2564960 TI - Horizontal transmission of hepatitis B virus. AB - Meta-analysis of seroprevalence surveys shows that horizontal transmission of hepatitis B virus (ie, that occurring without apparent parenteral, sexual, or perinatal exposure) is common in areas endemic for the virus. It occurs especially in pre-adolescent children. In developed countries, where endemicity of hepatitis B virus is low, horizontal transmission (probably via saliva or open wounds) may occur in households with a persistent carrier, but it is less efficient a means of infection than is sexual or perinatal transmission. Horizontal transmission also seems possible in pre-school day-care centres in developed countries, despite the small numbers of carriers in such places. PMID- 2564962 TI - Latitude and ischaemic heart disease. PMID- 2564963 TI - Stopping smoking and risk of ischaemic heart disease. PMID- 2564964 TI - Desmopressin and myocardial infarction. PMID- 2564965 TI - Antacids and drug trials for duodenal ulcer. PMID- 2564966 TI - Nafamostat to stabilise plasma samples taken for complement measurements. PMID- 2564967 TI - Malaria, hereditary elliptocytosis, and pyropoikilocytosis. PMID- 2564968 TI - Praziquantel for neurocysticercosis. PMID- 2564969 TI - Measurement of fetal peripheral perfusion with a pulse oximeter. PMID- 2564970 TI - Monitoring the anaesthetist. PMID- 2564971 TI - Ursodeoxycholic acid in primary biliary cirrhosis. PMID- 2564972 TI - Epstein-Barr virus infection and thyroid dysfunction. PMID- 2564973 TI - Clinical signs of pneumonia in children. PMID- 2564974 TI - Morbidity of very-low-birthweight infants. PMID- 2564975 TI - Aminophylline reversal of diazepam intoxication. PMID- 2564976 TI - Acute hepatic failure related to hepatitis A. PMID- 2564977 TI - Magnetic resonance imaging in diabetes insipidus. PMID- 2564978 TI - Suspected autochthonous kala-azar in Austria. PMID- 2564979 TI - Gene therapy. PMID- 2564980 TI - Mitochondrial DNA deletion in Pearson's marrow/pancreas syndrome. PMID- 2564981 TI - Yeast-derived hepatitis B vaccine and yeast sensitivity. PMID- 2564982 TI - Type III collagen deficiency. PMID- 2564983 TI - Hazards of oxygen treatment in the ambulance. PMID- 2564984 TI - Tyramine conjugation and antidepressants. PMID- 2564985 TI - Sicca syndrome in patients with Wegener's granulomatosis. PMID- 2564986 TI - Lupus anticoagulant induced by hydatidosis? PMID- 2564987 TI - Raynaud's symptoms improved by cold. PMID- 2564988 TI - Immunisation and general practice. PMID- 2564989 TI - Incentives for preventive medicine in general practice. PMID- 2564990 TI - Students and general practice. PMID- 2564991 TI - Diltiazem and heart block. PMID- 2564992 TI - Disabled child as income. PMID- 2564993 TI - NHS white-paper and medical education. PMID- 2564994 TI - Oncogenic viruses and cervical cancer. PMID- 2564995 TI - Cellular immune activation and erythropoiesis in gynaecological cancer. PMID- 2564996 TI - Rapid effect of oral limaprost in Raynaud's disease in childhood. PMID- 2564997 TI - Gender and survival in neuroblastoma. PMID- 2564998 TI - Radioimmunoassay of endothelin. PMID- 2565000 TI - Baldness and minoxidil. PMID- 2564999 TI - Asbestos-contaminated nappies and familial mesothelioma. PMID- 2565001 TI - Isothiazolinone sensitivity. PMID- 2565002 TI - Doppler ultrasound and early pregnancy failure. PMID- 2565003 TI - Dimethylformamide, metal dyes, and testicular cancer. PMID- 2565004 TI - Raised serum lipoprotein during treatment with lovastatin. PMID- 2565005 TI - Restraining the other shoulder. PMID- 2565006 TI - Hyperglycaemia, automatism, and insanity. PMID- 2565007 TI - AIDS in the UK and world wide. PMID- 2565009 TI - Persistence of cyclicity of the plasma dopamine metabolite, homovanillic acid, in neuroleptic treated schizophrenic patients. AB - The dopamine metabolite, homovanillic acid, decreases in concentration in plasma between 8:30 A.M. and 12:30 P.M. In patients with schizophrenia this cyclic change is attenuated by chronic neuroleptic treatment; however, if the 8 A.M. dose of neuroleptic is omitted, the decrease in level occurs. Presuming that neuroleptics attenuate the decline through a receptor mediated compensatory increase in dopamine release, it would appear that receptors are not fully occupied by neuroleptics even at therapeutically effective doses. The usual morning decrease in plasma cortisol levels was unaffected by neuroleptics. PMID- 2565008 TI - Prolactin (PRL) release-inhibiting properties of the alpha 2 adrenergic receptor antagonist idazoxan: comparison with yohimbine. AB - The effects of the alpha 2 adrenergic receptor antagonists yohimbine (YOH) and Idazoxan (ID) on secretion of PRL were compared in nonanesthetized male rats bearing permanent intraatrial cannulae for i.v. drug delivery and serial blood sampling. YOH induced a dose-related elevation of basal plasma PRL levels. ID had either no effect or a tendency to lower them and effectively inhibited stimulation of PRL secretion with morphine, 5-hydroxytryptophan (5HTP), quipazine or restraint stress. YOH at low doses did not alter the PRL secretory responses to these stimuli or enhanced them at the highest dose used (1.56 mg/kg). ID inhibited the PRL-stimulating, effect of 5HTP or morphine following inhibition of NE synthesis with FLA63 or pretreatment with clonidine. It also blocked the effect of quipazine in rats pretreated with prazosin. It is concluded that ID, in a complete contrast to YOH effectively inhibits PRL secretion. The inhibitory mechanism appears to be unrelated to its interaction with the alpha adrenergic receptors. PMID- 2565010 TI - [Using benzodiazepines for withdrawal from benzodiazepines?]. PMID- 2565011 TI - Drugs for cardiac arrhythmias. PMID- 2565012 TI - Biochemical events in spontaneous seizures in the Mongolian gerbil. AB - The Mongolian gerbil, with its spontaneous epileptiform seizures, was chosen as an experimental model of human epilepsy. Neurochemical parameters possibly related to the seizure process were studied. In the immediate seizure process amino acid profiles of cortex, hippocampus, and striatum were not different in seizuring animals when compared to seizure-resistance controls. Of two peptides analyzed, only somatostatin appeared elevated in the cortex 2 hr postictal (143 fmol/mg protein; controls, 123 fmol/mg protein); neuropeptide Y was not affected. A follow up of the time course of cyclic AMP and cyclic GMP showed significant elevations of both substances as a consequence of seizures. Most prominent was a 5.5-fold increase in cyclic GMP in the cerebellum 30 sec after seizure onset. PMID- 2565013 TI - Effect of trophic factors, released after hippocampal injury, on astroglial cell proliferation. AB - An increase in astrocyte mitogenic factors and in some specific astroglial enzymatic activities after neuronal injury has been observed. Our study is concerned with the effect of the intracerebral administration of ibotenic acid (IBO) into the rat hippocampus. IBO injection causes a selective degeneration of neurons while sparing afferent fibers. We observed a transient increase in glutamine synthetase activity, a well-known astroglial marker, reaching a peak at 9-15 days after injury in lesioned hippocampus. We investigated the presence of astrocyte mitogenic factors at various times after toxin injection. Crude extracts, prepared from lesioned hippocampi 4, 9, and 14 days after IBO injection, were tested for the ability to stimulate [methyl-3H]thymidine incorporation into rat astroglial cell cultures. Crude extracts prepared 9 and 14 days after IBO injection showed a higher mitogenic activity compared to extracts prepared 4 days after lesion. Mitogenic activity of injured brain extracts was suppressed by heat inactivation (100 degrees C for 10 min). PMID- 2565014 TI - Vecuronium bromide as an alternative to pancuronium bromide for neuromuscular blockade in the mechanically ventilated neonate. PMID- 2565015 TI - [Revascularization with the branched graft in middle aortic syndrome]. AB - A case of middle aortic syndrome which was thought to be the thoracoabdominal type of Takayasu's disease was successfully treated with the branched graft bypass. Patient was a 23 year-old woman with hypertension and abdominal pain. The preoperative angiography revealed aortic stenosis from the celiac axis to the left renal artery. The operative procedures were as follows; patient was positioned in supine with her left shoulder and arm raised. Eighth intercostal thoracotomy and midline laparotomy was performed with the thoracoabdominal incision. The branched graft was made previously with woven Dacron (phi 18 mm) and three EPTFEs (phi 8mm). The woven Dacron of the graft was used for the bypass from the descending thoracic aorta to the infrarenal abdominal aorta, and the branched EPTFEs of the graft were used for the bypasses to the common hepatic artery, the superior mesenteric artery and the right renal artery in this order. The bypasses were placed along the anatomical courses in the retroperitoneal space. Postoperatively, the blood pressure dropped and the abdominal pain disappeared. The plasma renin activity decreased and the renal function improved. Two months after operation the bypasses were patent by the angiography and now six months after operation she has returned to her social life healthily. The approach to the aorta and its abdominal branches by thoracotomy and laparotomy and bypass with the three branched graft was useful for middle aortic syndrome. PMID- 2565016 TI - [Wound healing, blood coagulation and fibrinolysis during operations involving gastric cancer surgery]. AB - Serial changes of FPA, FPB beta 15-42, FN, XIIIa, and alpha 2PI were investigated for the study on wound healing, blood coagulation, and fibrinolysis during gastric cancer surgery. For a control, we compared the preoperative values with the postoperative ones. These results also were compared with the values in healthy volunteers and in patients with cholelithiasis or myoma uteri. Our findings were as follows; 1) Compared with the control values, a statistically significant elevation of FPA, FPB beta 15-42 and FPA/FPB beta 15-42 ratios in patients with gastric cancer was noticed after operation. 2) Compared with the control values, a statistically marked decrease of FN, XIIIa and alpha 2PI in patients with gastric cancer was observed after operation. 3) The preoperative FPA and FPB beta 15-42 levels of gastric cancer patients were appreciably greater than those of normal healthy volunteers. Compared with patients with cholelithiasis or myoma uteri, however, the only preoperative FPA of gastric cancer patients showed significantly high levels. 4) FN and alpha 2PI revealed a notable positive correlation. These results suggest (1) increase of coagulation activity (thrombin formation) in gastric cancer patients; (2) increase of intravascular coagulation and fibrinolytic activity (thrombin and plasmin formation) during gastric cancer surgery; and, (3) depression of FN, XIIIa and alpha 2PI during surgery was due to sequestration at the site of tissue injury. PMID- 2565017 TI - Preparation and utility of a radioiodinated analogue of daunomycin in the study of multidrug resistance. AB - Anthracyclines are an important class of cytotoxic drugs that are frequently used in cancer chemotherapy, especially in acute leukemia. The pharmacokinetics and disposition of these compounds in whole animals and in cells have been studied employing 3H-labeled forms. However, their usefulness is limited by their low specific activities and the low energy of 3H. Therefore, we have labeled daunomycin using 125I-Bolton-Hunter reagent. The resultant anthracycline analogue, iodomycin, has a specific activity of approximately 2000 Ci/mmol. Although this compound was 10-fold less toxic to normal cells than daunomycin, multidrug-resistant cells were cross-resistant to it. Like other drugs to which these cells are cross-resistant, its accumulation by them was greatly reduced, compared with drug-sensitive cells. We have also utilized this compound in photoaffinity labeling experiments to identify its target in multidrug-resistant cells. We observed the specific binding of iodomycin to P-glycoprotein in membrane vesicles as well as in intact cells, thereby directly demonstrating that this protein specifically binds anthracyclines as well as Vinca alkaloids. PMID- 2565018 TI - Sensitive detection and schizodeme classification of Trypanosoma cruzi cells by amplification of kinetoplast minicircle DNA sequences: use in diagnosis of Chagas' disease. AB - Amplification of DNA sequences from the kinetoplast minicircle DNA was employed as a method for the detection and classification of small numbers of Trypanosoma cruzi cells. Two overlapping fragments from the conserved 120 bp minirepeat regions of the minicircle DNA and one fragment covering the adjacent variable regions were amplified. The minimal amount of minicircle DNA required to detect a product by hybridization with an oligonucleotide probe was 0.015 fg, which represents approximately 10 molecules or 0.1% of the minicircle DNA component of a single cell. The amplification worked equally well with kDNA from several strains of T. cruzi and did not occur with kDNA from several other kinetoplastids. kDNA recovered from less than 10 trypanosomes in whole blood could be used as a template for amplification; the presence of a several billion fold excess of human DNA had no effect on the amplification process. Schizodeme analysis by hybridization with specific oligonucleotides or by direct restriction enzyme digestion could be performed on the amplified fragments representing the minicircle conserved region or variable regions. This method should prove useful as a rapid, specific and sensitive assay for Chagas' disease in chronic patients as well as for epidemiological studies of infected animals and insects. PMID- 2565019 TI - Antigenic variation. Modulating bacterial virulence. PMID- 2565020 TI - Primary and transplantable hepatomas induced by aflatoxin B1 in hypothyroid rats. AB - The influence of hypothyroidism on AFB1 carcinogenesis in inbred Wistar rats was studied. The primary AFB1 hepatocarcinogenesis was significantly delayed and reduced. The transplantable hepatomas showed prolonged latency periods in rats of both sexes, and in males decreased tumor weights and a lower number of lung metastases were observed. The level of GGTP in serum and liver paralleled the advancement of morphological hepatic lesions, and the activity of serum GGTP in hypothyroid bearers of the transplantable hepatoma was lower in females than in males. It was concluded that the lack of thyroid hormones during the latency period of primary and transplantable hepatomas is decisive for delayed AFB1 carcinogenesis. PMID- 2565021 TI - Complex biochemical analysis of human breast tumor tissue. AB - The quantitative biochemical analysis of tissue specimens from 76 human breast carcinomas consisted of examination for cytosolic estrogen receptors (cER), nuclear estrogen receptors (nER), progesterone receptors (PgR), 1,25 dihydroxycholecalciferol receptors (DR), carcinoembryonic antigen (CEA), alpha lactalbumin (aLA), and gamma-glutamyl transferase (gGT). The highest incidence was found in CEA (76%), DR (70%), and aLA (62%). There was a high percentage of tumors containing only DR, in contrast to the tumors containing only cER or PgR. The simultaneous occurrence of DR and CEA was considerably high (61%). No statistically significant differences were observed in these biochemical parameters in relation to the grade of differentiation of the tumors. The values of aLA in tumors that invaded lymphatic or blood vessels were lower as compared to those tumors that invaded adipose or connective tissues. The level of statistical significance of this difference was close to 5%, the differences in other parameters were statistically insignificant. For prognosis assessed at the time of surgery, after a 2-3-year follow-up of 36 patients the level of gGT in the tumor seems to be the most promising prognostic factor. The values of gGT were significantly lower in those patients whose tumors were in progression during this time. The significance of nER and aLA was also taken into consideration. PMID- 2565022 TI - Immunophenotypic analysis of the inflammatory infiltrate in ocular cicatricial pemphigoid. Further evidence for a T cell-mediated disease. AB - Ocular cicatricial pemphigoid (OCP) is characterized by the deposition of immunoglobulin and complement along the conjunctival epithelial basement membrane zone (BMZ). In order to further elucidate the cellular populations of the local inflammatory infiltrates, the authors used a panel of monoclonal antibodies in cryostat tissue sections to delineate T cell subsets, B lymphocytes, dendritic cells, and macrophages in six patients with OCP. In comparison with matched controls of the epibulbar conjunctiva, the authors discovered a threefold increase in T lymphocytes within the epithelium and a 20-fold increase within the substantia propria. In contrast with the normal-standing population of conjunctival T lymphocytes, there were activated interleukin 2 receptor (IL-2R) positive lymphocytes in both the epithelium and the substantia propria. Macrophages were the second most common cells in the substantia propria, accounting for 12.7% of the mononuclear population--a threefold increase over the normal percentage. B cells and plasma cells, normally absent from epibulbar conjunctiva, were the next most prominent populations, constituting 6.9 and 4.6%, respectively, of all mononuclear cells. Dendritic cells which process antigen locally constituted only 1.2% of the mononuclear cell population, but were increased 25-fold over normal controls. By elaborating cytokines that promote fibroplasia, the T cells in OCP may be effector cells along with macrophages and other inflammatory cells in bringing about scarification of the substantia propria, and may furthermore be responsible for an immunoregulatory defect that allows local B lymphocytes to produce autoantibodies to the BMZ. PMID- 2565023 TI - Polymerase chain reaction: replication errors and reliability of gene diagnosis. AB - The impact of replication errors on the reliability of polymerase chain reaction (PCR) data is studied theoretically. Practical applications of our results to RFLP analysis and oligonucleotide probing confirm that for practical purposes replication errors can be neglected if a large number of starting templates (e.g. 100,000) is being used. For single locus analysis in single cells, however, the probability of false diagnosis due to such errors is of the order of 1 percent. PMID- 2565024 TI - Human non-histone chromosomal protein HMG-17: identification, characterization, chromosome localization and RFLPs of a functional gene from the large multigene family. AB - The multigene family of chromosomal protein HMG-17 is the largest known human retropseudogene family. A functional gene was identified and isolated by screening cDNA-selected genomic clones with a set of 5 oligonucleotides whose sequence corresponded to regions in which the sequence of the retropseudogenes differed from that of the cDNA and which did not span previously identified exon/intron junctions. A 7195 bp genomic fragment containing 6 exons, ranging in size from 30 to 817 bp, two of which encode the entire DNA binding domain of the protein, was sequenced. The gene has features which are typical to "housekeeping" genes and is characterized by a very high content of G + C residues in a 1.4 kb fragment starting 500 bp from the cap site and by an "HTF" island in the 5' region. Transcriptional regulatory signals, exon/intraon boundaries and features characteristic of "housekeeping" genes are evolutionary conserved between the human and chicken genes. The HMG-17 gene was localized to human chromosome 1p12 34. RFLP's useful for further mapping were detected. The experimental evidence presented leads to the assumption that the gene characterized is the only functional human HMG-17 gene. PMID- 2565025 TI - Two RFLPs associated with the human endogenous retroviral element S71 on chromosome 18q21. PMID- 2565026 TI - A rare TaqI RFLP immediately 3' of the HEXB gene on chromosome 5. PMID- 2565027 TI - A new BglII RFLP at the epidermal growth factor receptor locus (erbB-1) on chromosome 7. PMID- 2565028 TI - RFLP for HindIII at the Duchenne muscular dystrophy gene. PMID- 2565029 TI - An anonymous single-copy clone, p55-B1, from chromosome 17 identifies a TaqI RFLP [HGM9 no. D17S86]. PMID- 2565031 TI - RFLP for Sst I (Sac I) at the human plasminogen activator inhibitor type 1 (PAI 1) gene locus. PMID- 2565030 TI - Human T-cell receptor CD3-epsilon (CD3E)/TaqI DNA polymorphism. PMID- 2565032 TI - Hinf I polymorphism 3' to the human beta-globin gene detected by the polymerase chain reaction (PCR). PMID- 2565033 TI - Analysis of B-G and immune response genes in the Iowa State University S1 chicken line by hybridization of sperm deoxyribonucleic acid with a major histocompatibility complex class II probe. AB - Sperm DNA was isolated from chickens of the Iowa State University S1 line. Birds were from sublines selected for B-G antigen, humoral immune response to glutamic acid-alanine-tyrosine (IrGAT), and response to Rous sarcoma virus-induced (RSV) tumors. The DNA was digested with restriction enzymes and subjected to Southern blot analysis with a DNA probe specific for a class II gene of the chicken major histocompatibility complex (MHC). Restriction fragment length polymorphisms (RFLP) associated with B-G antigen type were found after digestion of DNA with three enzymes, PvuII, BglII, or Sau3A, out of a total of 15 tested. No RFLP were shown to be associated with IrGAT or RSV type. This study shows that RFLP analysis of DNA may be a useful addition to or alternative to serological evaluation of MHC haplotype in the chicken. PMID- 2565034 TI - Amplification of the proto-neu oncogene facilitates oncogenic activation by a single point mutation. AB - To determine whether the amplification of the proto-neu oncogene (also called c erbB-2) plays a role in tumorigenicity, we previously generated an NIH 3T3 transfectant (DHFR/G-8) that carried the amplified proto-neu gene. The DHFR/G-8 cells exhibited normal morphology. Their growth curve was similar to that of NIH 3T3 cells but was different from that of the B104-1 cell, and NIH 3T3 transfectant that carries the activated neu oncogene. When injected into nude mice, B104-1 cells produced tumors within 2 weeks, whereas the DHFR/G-8 cells did not produce tumors until 3 months after injection, and the NIH 3T3 cells did not produce any tumors even after 3 months. The tumors produced by the injection of the DHFR/G-8 cells were excised and grown in culture. The cells derived from the tumors were of transformed morphology and highly tumorigenic. The DNAs from the tumor cells were transfected into NIH 3T3 cells. The transfection resulted in foci on the NIH 3T3 monolayer. Southern analysis indicated that the foci derived from the transfection contained the neu gene. Using oligonucleotides as probes, the neu gene in the foci was found to carry a single-point mutation identical to the one previously found in the rat neuroblastoma and glioblastoma induced by the ethylnitrosourea. We conclude that the DNA region encoding the transmembrane domain of neu is a hot spot for converting the proto-neu gene into an activated oncogene and that amplification of the proto-neu gene facilitates mutation of the hot spot. PMID- 2565036 TI - Allozymes of glucose-6-phosphate isomerase differentially modulate pentose-shunt metabolism in the sea anemone Metridium senile. AB - We tested the hypothesis that kinetic differences among allelic variants of glucose-6-phosphate isomerase (GPI; D-glucose-6-phosphate ketol-isomerase, EC 5.3.1.9) from the sea anemone Metridium senile differentially modulate glucose metabolism at the glycolysis-pentose-shunt branch point. Fractional contribution of pentose shunt and absolute flux of glucose in glycolysis were measured in fasted or fed anemones acclimated to 5 degrees C or 15 degrees C. When fed, anemones of genotype Gpiss routed a greater fraction of glucose through the shunt than did Gpiff anemones; the effect was more pronounced at 5 degrees C than at 15 degrees C. This confirms predictions from kinetic and population data and is consistent with thermal selection maintaining the variation. Relative levels of shunt metabolism increased at 5 degrees C, compared with 15 degrees C, in fed anemones regardless of genotype, but the proportion of glucose metabolized by the pentose shunt was unchanged by temperature in fasted anemones. Glucose flux through the shunt was constant at approximately 5 pmol.mg-1.hr-1 in fed anemones at 5 degrees C and 15 degrees C and in fasted anemones at 15 degrees C, indicating apparently near-perfect thermal acclimation of the absolute flux of glucose through the shunt in fed, but not in fasted, anemones. Rates of glucose oxidation and flux through the shunt in freshly collected anemones were similar to those of anemones fed and acclimated at 15 degrees C in the laboratory. If these differences affect organismal-level processes, Gpi variation could contribute to Darwinian fitness in thermally varying environments. PMID- 2565035 TI - Characterization of the human 5-lipoxygenase gene. AB - The gene for human 5-lipoxygenase has been isolated from three different bacteriophage genomic libraries and a genomic cosmid library. The gene spans greater than 82 kilobases and consists of 14 exons. The size range for the exons is 82-613 base pairs, whereas that for the introns is approximately 200 bp to greater than 26 kb. A major site of transcription initiation in leukocytes was mapped to a thymidine residue 65 base pairs upstream of the ATG initiation codon by nuclease S1 protection and primer extension experiments. Other potential minor initiation sites were found. The putative promoter region contains no TATA and CCAAT sequences in the expected positions upstream of the major transcription initiation site but contains multiple GC boxes within a (G + C)-rich region, as does the immediate 5' region of the first intron. Characteristics common to the 5' end of the human 5-lipoxygenase gene and the promoter regions of the housekeeping genes raise important questions concerning the regulation of 5 lipoxygenase gene expression. PMID- 2565037 TI - Two-dimensional DNA fingerprinting of human individuals. AB - The limiting factor in the presently available techniques for the detection of DNA sequence variation in the human genome is the low resolution of Southern blot analysis. To increase the analytical power of this technique, we applied size fractionation of genomic DNA restriction fragments in conjunction with their sequence-dependent separation in denaturing gradient gels; the two-dimensional separation patterns obtained were subsequently transferred to nylon membranes. Hybridization analysis using minisatellite core sequences as probes resulted in two-dimensional genomic DNA fingerprints with a resolution of up to 625 separated spots per probe per human individual; by conventional Southern blot analysis, only 20-30 bands can be resolved. Using the two-dimensional DNA fingerprinting technique, we demonstrate in a small human pedigree the simultaneous transmission of 37 polymorphic fragments (out of 365 spots) for probe 33.15 and 105 polymorphic fragments (out of 625 spots) for probe 33.6. In addition, a mutation was detected in this pedigree by probe 33.6. We anticipate that this method will be of great use in studies aimed at (i) measuring human mutation frequencies, (ii) associating genetic variation with disease, (iii) analyzing genomic instability in relation to cancer and aging, and (iv) linkage analysis and mapping of disease genes. PMID- 2565038 TI - Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms. AB - We developed mobility shift analysis of single-stranded DNAs on neutral polyacrylamide gel electrophoresis to detect DNA polymorphisms. This method follows digestion of genomic DNA with restriction endonucleases, denaturation in alkaline solution, and electrophoresis on a neutral polyacrylamide gel. After transfer to a nylon membrane, the mobility shift due to a nucleotide substitution of a single-stranded DNA fragment could be detected by hybridization with a nick translated DNA fragment or more clearly with RNA copies synthesized on each strand of the DNA fragment as probes. As the mobility shift caused by nucleotide substitutions might be due to a conformational change of single-stranded DNAs, we designate the features of single-stranded DNAs as single-strand conformation polymorphisms (SSCPs). Like restriction fragment length polymorphisms (RFLPs), SSCPs were found to be allelic variants of true Mendelian traits, and therefore they should be useful genetic markers. Moreover, SSCP analysis has the advantage over RFLP analysis that it can detect DNA polymorphisms and point mutations at a variety of positions in DNA fragments. Since DNA polymorphisms have been estimated to occur every few hundred nucleotides in the human genome, SSCPs may provide many genetic markers. PMID- 2565039 TI - Mitotic recombination of chromosome 17 in astrocytomas. AB - Allelic combinations at seven loci on human chromosome 17 defined by restriction fragment length polymorphisms were determined in tumor and normal tissues from 35 patients with gliomas. Loss of constitutional heterozygosity at one or more of these loci was observed in 8 of the 24 tumors displaying astrocytic differentiation and in the single primitive neuroectodermal tumor examined. The astrocytomas showing these losses included examples of each adult malignancy grade of the disease, including glioblastoma (malignancy grade IV), and seven of them demonstrated concurrent maintenance of heterozygosity for at least one chromosome 17 locus. Determination of allele dosage together with the genotypic data indicated that the tumor chromosomes 17 were derived by mitotic recombination in 7 of the 9 cases with shared homozygosity of the region 17p11.2 pter in all cases. In contrast, tumors of oligodendrocytic, ependymal, or mixed cellular differentiation did not exhibit loss of alleles at any of the loci examined. These data suggest that the somatic attainment of homozygosity for loci on chromosome 17p is frequently associated with the oncogenesis of central nervous system tumors, particularly those showing solely astrocytic differentiation, and that mitotic recombination mapping is a useful approach towards the subregional localization of a locus whose rearrangement is involved in this disease. PMID- 2565040 TI - Elevated dynorphin in the hippocampal formation of aged rats: relation to cognitive impairment on a spatial learning task. AB - Radioimmunoassay revealed increased dynorphin A(1-8)-like immunoreactivity [dynA(1-8)LI] in the aged rat brain. Among a number of brain regions examined, an age-related dynA(1-8)LI elevation was found only in the hippocampal formation and frontal cortex. Moreover, the increase in dynA(1-8)LI in the aged hippocampus was associated with a decline in spatial learning ability: dynA(1-8)LI distinguished aged rats that were behaviorally impaired from aged cohorts that learned the spatial task as rapidly as younger animals. Northern blot hybridization using a 32P-labeled complementary RNA probe encoding rat prodynorphin indicated that the abundance of prodynorphin mRNA was also significantly increased in the hippocampal formation of aged rats with identified spatial learning impairments. PMID- 2565042 TI - Anatomical plasticity of synapses in the lamina of the optic lobe of the fly. AB - Insects are frequently assumed to have hard-wired nervous systems that fail to demonstrate functional plasticity. We have produced changes in synaptic frequency, and analysed their developmental time course, dynamics and reversibility, in the lamina underlying the compound eye of the fly, by exposing young adults to different visual stimuli. The class of synapse examined feeds back from L2, one of the monopolar cells found in each lamina cartridge, to photoreceptor terminals; each site is a synaptic dyad marked by the presence of a few, round vesicles surrounding a T-shaped presynaptic ribbon and, in the photoreceptor, by a subsynaptic vacuole. In control adult flies reared in normal room lighting, the frequency of synaptic profiles scored in micrographs of single sections initially increased until one day post-eclosion (E + 1), but declined thereafter. Frequencies measured in left and right eyes of the same control animals were closely matched. Experimental flies were put for one to two days into an integrating sphere illuminated continuously with square-wave, 25 Hz green light. They had one eye occluded, so providing control comparisons between flicker-reared (FR) and occluded (dark-reared, DR) eyes within the same animal. The DR eyes invariably (n greater than 22) had higher frequencies of synaptic profiles than those seeing light, regardless of age or the period of light exposure, although the detailed relative effects of FR and DR depend upon the age of the animal. The evidence suggests that exposure to light actively depresses synaptic frequency and increases its variability. The greatest difference (30%) achieved was at two to four days after eclosion and there was no difference beyond six days, so demarcating a prospective sensitive period. Rearing in DC light was equally effective as FR, so visual contrasts per se are apparently inessential. Frequency values can change rapidly. During the first 24 h post eclosion, DR resulted in new synapses adding to L2's complement of 25-35 at a maximum rate of 4 per 6 h, whereas light exposure caused a frequency decrease after as little as 6 h. Alternating 24 h periods of light and dark during the first two days produced reversible synaptic frequency changes. Individual synaptic contacts enlarge with age but not significantly with different visual experiences. The decrease in frequency of synaptic profiles with age thus actually underestimates the true decrease in synaptic number, whereas the altered synaptic frequencies seen after differential exposure represent true differences in synaptic number.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2565043 TI - [Imaging diagnosis at the threshold of the 21st century. A conference in honor of Antonio Toti. The Academy of Sciences and Medico-Surgical Society of Ferrara (16 November 1988)]. PMID- 2565041 TI - Regulation of ATP-sensitive K+ channels in insulinoma cells: activation by somatostatin and protein kinase C and the role of cAMP. AB - The actions of somatostatin and of the phorbol ester 4 beta-phorbol 12-myristate 13-acetate (PMA) were studied in rat insulinoma (RINm5F) cells by electrophysiological and 86Rb+ flux techniques. Both PMA and somatostatin hyperpolarize insulinoma cells by activating ATP-sensitive K+ channels. The presence of intracellular GTP is required for the somatostatin effects. PMA- and somatostatin-induced hyperpolarization and channel activity are inhibited by the sulfonylurea glibenclamide. Glibenclamide-sensitive 86Rb+ efflux from insulinoma cells is stimulated by somatostatin in a dose-dependent manner (half maximal effect at 0.7 nM) and abolished by pertussis toxin pretreatment. Mutual roles of a GTP-binding protein, of protein kinase C, and of cAMP in the regulation of ATP sensitive K+ channels are discussed. PMID- 2565044 TI - Evaluation of total plasma bile acid concentrations for the diagnosis of hepatobiliary disease in horses. AB - Plasma bile acid concentrations were measured in normal horses. There was no diurnal variation in values, and age and sex had no effect. There was no significant difference between serum and plasma bile acid concentrations in clinically normal horses. Plasma bile acids were stable on storage for one month at -20 degrees C. The total plasma bile acid concentrations together with total and direct bilirubin concentrations and plasma activities of aspartate aminotransferase, glutamate and iditol dehydrogenase were evaluated in horses with various types of hepatobiliary disease (hepatic necrosis, lipidosis, neoplasia and cirrhosis), gastrointestinal disease, cardiovascular, orthopaedic and various other conditions not affecting the liver. Total plasma bile acids together with plasma glutamate and iditol dehydrogenase activities were the best indicators of liver disease. Total plasma bile acid concentrations were the most sensitive indicator of a wide variety of hepatic diseases but alone were unhelpful in differential diagnosis and were of more value when combined with the other tests of hepatic disease. PMID- 2565045 TI - [3d consensus conference on intensive care and emergency medicine. Care of severe acute asthma crisis in adults]. PMID- 2565046 TI - Heritable allele-specific differences in amounts of apoB and low-density lipoproteins in plasma. AB - Low-density lipoprotein (LDL) concentrations correlate with risk of coronary heart disease, and genetic variation affecting LDL levels influences atherosclerosis susceptibility. The principal LDL protein is apolipoprotein B (apoB); apoB is not exchangeable between lipoprotein particles and there is only one apoB per LDL particle. Plasma LDL therefore consists of two populations, one containing apoB derived from the maternal and one from the paternal apoB alleles. Products of the apob gene with high or low affinity for the MB-19 monoclonal antibody can be distinguished, and this antibody was used to identify heterozygotes with allele-specific differences in the amount of apoB in their plasma. A family study confirmed that the unequal expression phenotype was inherited in an autosomal dominant manner and was linked to the apob gene locus. Significant apoB genetic variation affecting plasma LDL levels may be more common than previously appreciated. PMID- 2565047 TI - Allelotype of colorectal carcinomas. AB - To examine the extent and variation of allelic loss in a common adult tumor, polymorphic DNA markers were studied from every nonacrocentric autosomal arm in 56 paired colorectal carcinoma and adjacent normal colonic mucosa specimens. This analysis was termed an allelotype, in analogy with a karyotype. Three major conclusions were drawn from this analysis: (i) Allelic deletions were remarkably common; one of the alleles of each polymorphic marker tested was lost in at least some tumors, and some tumors lost more than half of their parental alleles. (ii) In addition to allelic deletions, new DNA fragments not present in normal tissue were identified in five carcinomas; these new fragments contained repeated sequences of the variable number of tandem repeat type. (iii) Patients with more than the median percentage of allelic deletions had a considerably worse prognosis than did the other patients, although the size and stage of the primary tumors were very similar in the two groups. In addition to its implications concerning the genetic events underlying tumorigenesis, tumor allelotype may provide a molecular tool for improved estimation of prognosis in patients with colorectal cancer. PMID- 2565048 TI - [Development of infection with human immunodeficiency virus]. PMID- 2565049 TI - [Torture--helper and hangman?. Interview by Bjorn Arild Ostby]. PMID- 2565050 TI - [Tourette syndrome. New aspects on pharmacological treatment]. AB - Tourette syndrome is now widely accepted as a neurological disorder characterized by chronic multiple tics and uncontrollable sounds. The exact etiology of Tourette syndrome is still unknown, although an organic etiology is suspected. Hypotheses of the pathophysiologic mechanism of Tourette syndrome have generally included a component of neurotransmitter system "imbalance". Dopaminergic, cholinergic, serotoninergic, noradrenergic and peptidergic neurotransmitters have all been proposed as components of this "imbalance". The two most utilized agents, haloperidol and pimozide, decrease central dopaminergic activity by blocking dopamine receptors and have been shown to improve the symptomatology of Tourette syndrome. However, adverse effects limit their usefulness and new drugs that have fewer adverse effects and greater efficacy are needed. Some of the new therapeutic agents evaluated in Tourette syndrome are mentioned. The purpose of this paper is to provide physicians with guidelines for the pharmacological treatment of Tourette syndrome, based on the present available literature. PMID- 2565051 TI - Vertebrate homeodomain proteins: families of region-specific transcription factors. AB - Vertebrate homeodomain proteins are transcription factors whose genes can be isolated via a conserved DNA-binding domain called the homeobox. We review recent studies suggesting that one function of these genes is the early subdivision of the embryo along the antero-posterior axis into 'fields' of cells with different developmental potential. PMID- 2565052 TI - Clinical collection and use of peripheral blood stem cells in pediatric patients. AB - Peripheral blood stem cells have been used in lieu of marrow as autologous cellular support after marrow-ablative radio/chemotherapy. Autologous marrow support allows the use of higher, more effective doses of therapy. Peripheral blood stem cells (PBSC) offer the potential for this form of "transplant" in patients with marrow involved with disease. PBSC use also allows the avoidance of marrow harvest under general anesthesia. Previous reports of PBSC use have been limited to adults. This article describes peripheral blood stem cell collection in children, and documents the successful use of such cells in a 21/2-year-old boy and a 3-year-old girl with progressive stage IV neuroblastoma involving the marrow. Collections were performed using a Fenwal CS3000 blood cell processor primed with fresh frozen plasma and red cells, using a low flow rate to avoid citrate toxicity. The transplant was performed using collections that were negative or low in neuroblastoma cells (as detected by anti-neuron-specific enolase and antineuroblastoma monoclonal antibodies) after preparation with high dose cyclophosphamide, etoposide, and cisplatin. Posttransplant recovery was uneventful, and engraftment was comparable to that in 4 patients treated with a similar preparative regimen followed by infusion of autologous marrow. Using careful technique, PBSC support in lieu of marrow may be a viable treatment modality in pediatric neuroblastoma or other solid tumors, particularly when the marrow is involved with disease. PMID- 2565053 TI - Signaling via the T-cell antigen receptor heterodimer and the CD2 antigen: a novel, synergistic pathway for activation of T-cells. PMID- 2565054 TI - The T lymphocyte CD2 antigen--genetic and functional studies. PMID- 2565055 TI - Prolongation of cardiac allograft survival by pretransplant administration of cyclosporine and donor-specific dendritic cells. PMID- 2565057 TI - Reassessment of the role of class II antigens in skin graft rejection. PMID- 2565056 TI - CD4+ lymphocytes play a dominant role in murine xenograft rejection. PMID- 2565058 TI - DNA typing of HLA class II antigens is informative in kidney transplantation. PMID- 2565059 TI - Matching for HLA-D region and T-cell antigen receptor gene restriction: fragment length polymorphisms in renal transplantation. PMID- 2565060 TI - Are OKT3 binding sites on T cells modulated or merely converted (blindfolded) by OKT3 during therapy? PMID- 2565061 TI - Serum dipeptidyl peptidase IV in cardiac transplant recipients. PMID- 2565062 TI - Functional adaptation of small intestinal mucosa after syngeneic and allogeneic orthotopic small bowel transplantation. PMID- 2565063 TI - Autologous blood stem cell transplantation. PMID- 2565064 TI - Hemopoietic growth factors and marrow transplantation: an overview. AB - Current clinical trials suggest that use of G-CSF and GM-CSF is likely to benefit marrow transplant patients by significantly reducing the duration of post transplant leukopenia. It remains to be determined whether the use of megakaryocyte-active growth factors (e.g., Multi-CSF) will prove of value in elevating platelet levels in such patients. PMID- 2565065 TI - Allogeneic bone marrow transplantation in newborn mice treated pre- and post natally with anti-THY 1.2 monoclonal antibody. PMID- 2565066 TI - Fate of donor T cells in systemic graft-versus-host reaction. PMID- 2565067 TI - Autologous peripheral blood stem cell transplantation: analysis of autografted cells and lymphocyte recovery. PMID- 2565068 TI - Unmatched stem cell transplantation as a possible alternative to bone marrow transplantation. PMID- 2565070 TI - Immunological clearance of 75Se-labelled Trypanosoma brucei in goats. AB - An experiment was conducted to determine, under different conditions, the capacity of young adult East African goats to eliminate intravenously inoculated [75Se]selenomethionine-labelled Trypanosoma brucei from the bloodstream. Over 80% of labelled trypanosomes, preincubated for 1 h in inactivated normal goat serum, were detectable in the circulation 1 h after inoculation into normal goats. By contrast, after incubation in serum from goats which had been immunised against the homologous trypanosome clone, parasites were largely removed from the bloodstream within 5 min after inoculation. When the goats were necropsied 1 h after the inoculation of radiolabelled trypanosomes, 50% of the injected activity was found in the liver and lungs, the contribution of each organ being dependent to some extent on whether the inoculum was via a mesenteric or the jugular vein. The same result was obtained when labelled parasites were incubated in normal goat serum, and then inoculated into immunised goats; thus, rapid blood clearance occurred, and high activity was detected in the lungs and liver. The results confirm those of previous studies in laboratory mice in which the removal of trypanosomes from the circulation of an immune animal was achieved primarily by uptake of opsonised trypanosomes by elements of the mononuclear phagocytic system. PMID- 2565069 TI - [Current problems in the prevention of sudden cardiac death]. AB - Within the framework of the control of cardiovascular diseases the prophylaxis of the sudden heart death is an actual problem of health policy and medicine. Since the most frequent cause of the sudden heart death is the coronary heart disease, elements of the primary and secondary prophylaxis of the coronary heart disease are also of importance for the prevention of the sudden heart death. Central points of the prophylaxis of the sudden heart death are the improvement of the prehospital care and the shortening of the prehospital times of delay in acute myocardial infarction as well as the recognition, registration and care of risk persons. Nowadays the demand is existing more to fulfil these tasks in the daily practice. The actual possibilities of the medicamentous prophylaxis are analysed and discussed. More and more comprehensive knowledge about the prophylactic effect of the beta-receptor blockers as to the reduction of the frequency of the sudden hearth death is existing. They are most effectful in persons with a high risk particularly after an acute myocardial infarction. Though beta-receptor blockers shall cause changes of the serum lipids, on the basis of a more fundamental analysis of literature can be stated, since there is no clue to it, that beta-receptor blockers further the development of arteriosclerosis. On the contrary there are references for the fact that the development of arteriosclerosis is delayed. For this reason the demand has to be made to fully use the therapeutical potential of the beta-receptor blockers in the therapy of the chronic cardiovascular diseases and their complications.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565071 TI - Glossina morsitans morsitans: mortalities caused in adults by experimental infection with entomopathogenic fungi. AB - Various strains of the entomopathogenic fungi: Beauveria bassiana, Metarhizium anisopliae, Paecilomyces fumosoroseus and P. farinosus were found to be pathogenic for adult tsetse, Glossina morsitans morsitans but B. bassiana and M. anisopliae were the most pathogenic, often causing mortalities of up to 100%. Dose-mortality relationships were demonstrated for both B. bassiana and M. anisopliae and male tsetse were observed to be more susceptible to infection than females. Pure cultures of B. bassiana and M. anisopliae were isolated from haemolymph and fat body of tsetse previously exposed to fungal spores and hyphal fragments were often observed floating freely in the haemolymph. No increase in abortions was observed in female pregnant tsetse infected with B. bassiana and M. anisopliae and pupae produced by the infected females showed no increase in pupal mortality. Furthermore, when larvae were heavily dusted with spores of B. bassiana and M. anisopliae prior to pupation and incubation, no increase was observed in pupal mortality suggesting lack of fungal infection. PMID- 2565072 TI - Presence of circulating anodic antigen in serum of Schistosoma intercalatum infected patients from Gabon. AB - The presence of the schistosome circulating anodic antigen (CAA) in serum of Schistosoma intercalatum-infected patients from Gabon has been investigated using an enzyme-linked immunosorbent assay (ELISA). Blood samples were collected from 10 endemic controls, 29 patients which excreted viable S. intercalatum eggs in rectal mucosa and stool, six persons in which only non-viable eggs were found in rectal biopsy specimens and one person in which besides non-viable eggs a small number of viable eggs was found in the rectal biopsy specimen. CAA, a genus specific antigen, could be demonstrated in 58.6% of the patients with S. intercalatum eggs in their stools. In comparison to S. mansoni infections, very light infections (0.6 eggs per gram faeces) could be detected by the ELISA. A strong correlation between parasite burden (eggs per gram faeces) and antigen level (CAA-titer) was found (Spearman's rho = 0.65). Only one positive ELISA results was found in the group with solely non-viable eggs in rectal tissue. As no false positive results were detected for the negative controls, the present results suggest, in accordance with results earlier obtained for schistosomiasis mansoni, that only in active S. intercalatum infections is antigen demonstrable. PMID- 2565073 TI - Cutaneous leishmaniasis of the New World: diagnostic immunopathology and antigen pathways in skin and mucosa. AB - Non-specific chronic inflammation and/or granulomatous reaction are the main histopathological manifestations of cutaneous and mucocutaneous leishmaniasis of the New World. Plasma cell infiltration associated with collagen and vascular changes are data suggestive but not diagnostic of the disease. Specific diagnosis is only possible through demonstration of the parasite in the tissue examined. It is noteworthy that the parasites are usually scanty and difficult to demonstrate in the lesions. Biopsies from 40 patients with cutaneous or mucocutaneous leishmaniasis were examined using the immunofluorescence and immunoperoxidase techniques in order to demonstrate the parasite and/or antigen in the tissues. Nineteen biopsies showed non-specific chronic inflammation and 21 a granulomatous reaction. Parasites were found in 20% of the routine biopsies. The positivity through indirect immunofluorescence was 88.46% in frozen sections of fresh material and 89.28% in paraffin embedded tissue. The antigen positivity with the immunoperoxidase technique was 64.51%. Antigen was detected as amastigotes and also as diffuse material in the macrophage cytoplasm and adsorbed in the epithelial basement membrane and vessel walls. There was no difference in the positivity of antigen according to the type of inflammatory reaction. PMID- 2565074 TI - Fatty acid composition of amastigote and trypomastigote forms of Trypanosoma cruzi. AB - The fatty acid composition of total lipids from trypomastigote and amastigote forms of Trypanosoma cruzi and of Vero cells before and after parasite infection were analyzed by gas-liquid chromatography and mass spectrometry. Even-numbered, saturated, monoenoic and polyenoic acids ranging from C-12 to C-18 were characterized in both T. cruzi development stages. Significant changes in the fatty acid composition occurred during the T. cruzi life cycle. Oleic and linoleic acids were prominent in trypomastigote forms, whereas palmitic acid was the major fatty acid of amastigotes. Other differences include higher stearic acid and lower palmitoleic and linolenic acid levels as well as the absence of lauric acid in amastigotes as compared with trypomastigote forms. The fatty acid pattern of Vero cells before T. cruzi infection as compared with that after infection showed mostly qualitative differences. Linoleic and linolenic acids were observed only in T. cruzi infected cells. PMID- 2565075 TI - Effects of induction of anti-embryonation immunity on liver granulomas, spleen weight and portal pressure in mice infected with Schistosoma japonicum. AB - BALB/c mice sensitized with injections of viable immature Schistosoma japonicum eggs had significantly fewer and smaller granulomas in the liver, lower portal pressure and smaller spleens at D + 75 of infection compared to similarly infected unsensitized controls. The portal pressure and spleen weights of the mice sensitized with immature eggs were not different from uninfected unsensitized mice of similar ages at D + 75 of infection. The results strongly support our hypothesis that it should be possible to prevent serious hepatosplenic disease in schistosomiasis japonica by vaccination to induce anti embryonation immunity. PMID- 2565076 TI - Different sensitivities of rat and human red cells to exogenous Ca2+. AB - During an examination of the effects of shear and of the Ca2+ ionophore A23187 on Ca2+ entry into erythrocytes of rats and humans, we noted that rat erythrocytes were much more sensitive to Ca2+-induced hemolysis than the human cells. An examination of the effect of Ca2+ on transglutaminase, a cytosolic enzyme in the erythrocyte which cross-links membrane proteins and renders cells less deformable, demonstrated a correlation between enzyme activity and Ca2+-induced hemolysis. Both rat and human cells subjected to shear-induced Ca2+ entry exhibited increased enzyme activity and altered membrane protein SDS-PAGE patterns. Twenty micromolar A23187 with Ca2+ at concentrations above 80 microM caused hemolysis of rat erythrocytes. In contrast to human erythrocytes, under these conditions no membranes were recoverable from rat erythrocytes. At lower concentrations of Ca2+ (25 and 50 microM), however, rat erythrocytes maintained integrity, and exhibited enhanced transglutaminase activity and cross-linking of membrane proteins. The rat enzyme can be activated 30% by 10 microM Ca2+, while 50 microM Ca2+ was necessary to achieve a similar activation of the enzyme from human red blood cells. In studies of shear-stimulated Ca2+ uptake by erythrocytes the rat red cell enzyme was more readily activated. The SDS-PAGE pattern of rat red cell membranes after a 30 sec shear showed specific changes in protein banding, including the appearance of bands greater than 330 kDa. Changes in protein banding were also apparent in cytosolic proteins. This work supports the view that shear-induced Ca2+ entry activates transglutaminase that leads to cross linking of membrane components, a loss of cell integrity, and eventual cell death. PMID- 2565077 TI - Haplotype distribution of the human phenylalanine hydroxylase locus in Scotland and Switzerland. AB - RFLP haplotypes at the phenylalanine hydroxylase (PAH) locus were determined in 45 nuclear Caucasian families from Switzerland and Scotland. The RFLPs at the PAH locus are highly informative, and prenatal diagnosis is possible in 85% of the families studied. The data were combined with the profiles previously observed in the Danish population, in order to study the variation in RFLP haplotype distribution among European populations. A total of 22 different haplotypes were observed in Denmark, Switzerland, and Scotland. Fifteen and 19 haplotypes are associated with the normal (non-PKU) and with the mutant chromosomes, respectively. The haplotype distribution and the allele frequency of normal chromosomes remain constant between Denmark, Switzerland, and Scotland. However, both the haplotype distribution and allele frequencies of mutant chromosomes show significant variation between the three countries. Our results suggest there may be additional mutations in the PAH gene that cause PKU. PMID- 2565078 TI - Major-histocompatibility-complex gene markers and restriction-fragment analysis of steroid 21-hydroxylase (CYP21) and complement C4 genes in classical congenital adrenal hyperplasia patients in a single population. AB - The gene CYP21B, encoding the steroid 21-hydroxylase enzyme of adrenal steroid biosynthesis, has been mapped to the human major histocompatibility complex (MHC). Deficiency of this enzyme leads to congenital adrenal hyperplasia (CAH). We report the phenotypes of the HLA and complement C4 and Bf genes, which are closely linked to the CYP21B gene, together with a detailed analysis of the CYP21 and C4 RFLP, in 17 Finnish families with CAH. The RFLP analysis with six restriction enzymes suggested that, altogether, 35% of the affected chromosomes had a CYP21B + C4B gene deletion, 9% an obvious gene conversion of the CYP21B gene to a CYP21A-like gene, and 3% a CYP21A + C4B duplication. The remaining 53% gave the RFLP patterns also found in nonaffected chromosomes. We also found that a 14.0-kb EcoRI RFLP marker of the CYP21 genes was strongly associated with the presence of a short C4B gene, suggesting that some of the RFLP markers found with the CYP21 probe may actually derive from C4B gene polymorphism. Three particular MHC haplotypes, each with a characteristic RFLP pattern, were found in many unrelated families. These three haplotypes accounted for 59% of the affected chromosomes in our study group, the rest (41%) of the affected chromosomes being distributed among various subtypes. The results suggest that, within a single, well-defined population such as in Finland, only a few CYP21B gene defects may constitute a substantial part of the affected chromosomes. This finding will help in genetic studies of CAH in such populations. PMID- 2565079 TI - Identification and characterization of 23 RFLP loci by screening random cosmid genomic clones. AB - As part of our search for polymorphic DNA probes, we have screened cosmids from a human genomic DNA library for their ability to reveal RFLPs. A total of 101 randomly isolated cosmid clones were tested in Southern hybridizations for polymorphic band patterns. Fifty-four of these clones revealed RFLPs with one or more of nine restriction enzymes. Twenty-three of these clones have been further characterized and assigned to 10 different chromosomes by linkage analysis or by hybridization to panels of human-hamster hybrid cell lines. Fifteen of the probes have heterozygosities greater than or equal to .5. The relative efficiency of RsaI and PstI restriction enzymes in detecting polymorphism was different from results obtained with libraries constructed in bacteriophage vectors. Screening randomly selected cosmid probes is an efficient method for detecting RFLPs. PMID- 2565080 TI - An intronic region within the human factor VIII gene is duplicated within Xq28 and is homologous to the polymorphic locus DXS115 (767). AB - The genomic sequences recognized by the anonymous probe 767 (DXS115) are localized to two sites within Xq28. One site lies within intron 22 of the factor VIII gene (FBC). Physical mapping suggests that the second site lies within 1.2 megabases of the F8C gene. The RFLPs detected by 767 are located within the second site. Genetic data suggest that F8C and DXS115 are tightly linked (theta max = .04; Zmax = 8.30). Recombination events in meioses informative for DXS52 (St14), DXS115, and F8C suggest that DXS115 and F8C lie distal to DXS52. PMID- 2565081 TI - Isolation of additional polymorphic clones from the cystic fibrosis region, using chromosome jumping from D7S8. AB - The cystic fibrosis (CF) locus has been located, by both linkage analysis and physical mapping, to a 900-kb region of 7q22-31 flanked by D7S8 (J3.11) and D7S23 (XV-2c). Using a 100-kb general jumping library, we isolated two sequential jump clones, J31 and J29, to one side of the D7S8 region and one jump clone, J32, to the other side of D7S8, so that the total region covered is about 300 kb. Three new RFLPs were detected by J29 and J32. Using PFGE mapping and the three jump clones, we found it possible to orient D7S8 on the chromosome and, by linkage analysis, to further narrow the CF region by 100 kb. The orientation of D7S8 will be useful for directing the isolation of other jump clones toward the CF locus. Though the newly described RFLPs are in considerable linkage disequilibrium with D7S8 polymorphisms, they increase the informativeness of genetic markers in the D7S8 region and should be useful in prenatal diagnosis. PMID- 2565082 TI - Isolation of a new DNA marker in linkage disequilibrium with cystic fibrosis, situated between J3.11 (D7S8) and IRP. AB - A cosmid library of recombinants containing nonmethylated CpG sites for rare cutter restriction enzymes was used previously to isolate the gene IRP and four polymorphic DNA markers (pPT-3, pXV-2c, pCS.7, and pKM.19) which are close to and in linkage disequilibrium with the cystic fibrosis (CF) mutation. We have analyzed several new clones from the same library and have isolated a further cosmid, cNX.6d, which maps approximately 160 kb from CS.7, in the J3.11 direction. A DNA fragment (pMP6d-9) (D7S399) derived from cosmid cNX.6d detects a frequent polymorphism with MspI. Strong linkage disequilibrium between CF and MP6d-9 is found in European populations. Recombinations in two families suggest that CF is between the MspI polymorphic site recognized by pMP6d-9 and the polymorphism recognized by pJ3.11. The new marker is the closest, to date, to CF and will be useful for prenatal diagnosis and carrier testing. PMID- 2565083 TI - Genetic linkage of Beckwith-Wiedemann syndrome to 11p15. AB - Beckwith-Wiedemann syndrome (BWS), characterized by multiorgan developmental abnormalities and predisposition to cancer, usually occurs sporadically, but small apparently dominant pedigrees have been described. Since rare patients show varying karyotypic abnormalities on the short arm of chromosome 11, it has been suggested that BWS may be related to the Wilms tumor gene on 11p13 or, alternatively, to growth factor genes on 11p15. We performed genetic linkage analysis on two BWS kindreds, using RFLPs for loci on 11p. BWS was linked to the insulin gene (11p15.5), with an overall maximum lod score of 3.60 (recombination fraction = .00). Linkage to D11S16 (11p13) could be excluded for recombination fractions less than or equal to .03. These results suggest that BWS defines a tumor-predisposition gene on 11p15. PMID- 2565084 TI - X-linked severe combined immunodeficiency: localization within the region Xq13.1 q21.1 by linkage and deletion analysis. AB - X-linked severe combined immunodeficiency (SCID) (McKusick 30040; IMD4) is a disease of unknown pathogenesis characterized by severe and persistent infections from early in life that are due to absence of both cellular and humoral immune function. Although the disease has been provisionally mapped to proximal Xq, high lethality and lack of a carrier test have limited the number of scorable meioses. We performed linkage analysis in six new kindreds with X-linked SCID, using a random pattern of T-cell X inactivation to rule out the carrier state in at-risk women. Our linkage results, combined with analysis of Xq interstitial deletions, confirmed the regional assignment of X-linked SCID, narrowed the boundaries within which this locus lies to Xq13.1-q21.1, and established the locus order DXS159-(PGK1, SCID)-DXS72-DXS3, defining flanking markers for prenatal diagnosis and carrier testing. PMID- 2565085 TI - Localization of the genetic defect in multiple endocrine neoplasia type 1 within a small region of chromosome 11. AB - Multiple endocrine neoplasia type I (MEN-1), a Mendelian disorder with an autosomal dominant mode of inheritance, causes hyperplasia in the parathyroid glands and hyperplasia or neoplasm in the anterior pituitary gland and/or the pancreatic islets. The genetic defect responsible for MEN-1 in three families was recently mapped to the long arm of chromosome II by linkage between the MEN-1 locus and the gene for skeletal muscle glycogen phosphorylase (PYGM) at 11q13. We have constructed a genetic linkage map of seven markers in the vicinity of the MEN-1 locus that has allowed us to map more precisely the gene associated with MEN-1; the target region has been narrowed to about 12 cM. The closely linked markers will be useful also for identification of likely carriers in families in which an allele responsible for MEN-1 segregates. PMID- 2565086 TI - Cardiorenal effects of atrial natriuretic factor administration in congestive heart failure: natriuresis and diuresis without hemodynamic alterations. AB - The effects of low bolus dose (70 +/- 6 micrograms [mean +/- SEM]) atrial natriuretic factor (ANF) administration was assessed in 16 patients with chronic congestive heart failure. Measurements were made for at least 60 minutes before and after the dose of ANF. There was a significant increase in urine flow rate (0.81 +/- 0.06 to 1.81 +/- 0.23 ml/min, p less than 0.01), sodium excretion rate (56 +/- 14 to 80 +/- 23 microEq/min, p less than 0.01), fractional excretion of sodium (1.23 +/- 0.49 to 1.63 +/- 0.60 percent, p less than 0.01) and potassium excretion rate (35 +/- 7 to 42 +/- 6 microEq/min, p less than 0.02). However, no significant alterations in renal plasma flow or glomerular filtration rate were observed. Furthermore, there was no significant correlation between the change in urine flow rate or sodium excretion rate and the change in renal plasma flow or glomerular filtration rate, respectively. In addition, there was no significant effect on cardiac index, mean aortic or left ventricular filling pressures, or systemic vascular resistance. There also was no discernible relationship between the response to ANF and the baseline concentrations of plasma ANF, aldosterone, or plasma renin activity. Thus, in patients with congestive heart failure, low dose ANF boluses may produce an increase in urine flow rate and sodium excretion rate that is independent of renal plasma flow or glomerular filtration rate. This suggests a meaningful direct renal tubular effect of exogenous ANF in this setting. PMID- 2565087 TI - Analysis of proliferative grade using anti-PCNA/cyclin monoclonal antibodies in fixed, embedded tissues. Comparison with flow cytometric analysis. AB - Cell kinetic information is an important adjunct to histologically-based tumor classifications. Presently, cell kinetic data can be obtained from slide-based material only with monoclonal antibodies such as Ki-67, which require the use of frozen sections and cannot be applied to archival, paraffin-embedded material. Monoclonal antibodies have recently been generated to PCNA/cyclin, a 36 kd, S phase-associated nuclear protein. The authors investigated whether monoclonal antibody 19A2 could be used to identify proliferating cells within fixed, embedded tissue sections. Deparaffinized sections of 41 methacarn-fixed human tumors were immunostained with 19A2 using a streptavidin biotin immunoperoxidase system. A semiquantitative scoring system was used to evaluate the fraction of cells that were PCNA/cyclin-positive, and this score was compared with cell kinetic data obtained from parallel flow cytometric S-phase analysis that had been performed on fresh samples of the same tumors. While there was general agreement between the slide-based, antibody-derived and the flow cytometrically derived cell kinetic information, some discrepancies were observed. Some of the latter represented cases in which the anti-PCNA/cyclin antibody preparations demonstrated significant heterogeneity in the numbers of proliferating cells in different regions of the tumor. In other cases, a significant fraction of the positive cells corresponded to nontumor stromal and/or inflammatory cells. In these cases, the slide-based method provided more information about the tumor cell population than did the flow cytometry data. It is concluded that semiquantitative immunocytochemical analysis with anti-PCNA/cyclin antibodies may represent a simple, reproducible, yet powerful technique for the routine analysis of cell kinetic data in alcohol-fixed, paraffin-embedded tissue by the surgical pathologist. PMID- 2565088 TI - Inhibitory regulation of rat exocrine pancreas by peptide YY and pancreatic polypeptide. AB - Peptide YY (PYY) and pancreatic polypeptide (PP) have been shown to inhibit exocrine pancreatic secretion in vivo in a variety of species. This study evaluates the type of stimulation inhibited by PYY and PP by examining, in urethan-anesthetized rats, the inhibition of pancreatic secretion when stimulated to a comparable extent by cholecystokinin (CCK), 2-deoxy-D-glucose (2DG), bethanecol, and electrical vagal nerve stimulation. PYY at maximal infusion rates inhibited stimulation by CCK by 83%, bethanecol by 55%, and electrical nerve stimulation by 40%. The inhibition of CCK stimulation was half maximal at 250 pmol.kg-1.h-1. By contrast, PYY totally inhibited 2DG-stimulated secretion with half-maximal inhibition at 10 pmol. kg-1.h-1. PP acted similarly to PYY in inhibiting CCK and 2DG-stimulated pancreatic protein secretion but was fivefold weaker in each case. These findings indicate that PYY and PP have multiple actions but preferentially inhibit neurally mediated pancreatic secretion at a preacinar cell locus, possibly at a central site of action. PMID- 2565089 TI - Inhibition of gastric and pancreatic secretion in dogs by CGRP: role of somatostatin. AB - The coexistence of calcitonin gene-related peptide (CGRP) and somatostatin (SS) within the stomach and pancreas and the potent inhibitory effects of both peptides on exocrine secretions from these organs suggest that they are functionally related. To assess the potential role of SS in the mediation of CGRP action, the effects of intravenous human CGRP (64, 132, and 264 pmol.kg-1.h-1) and somatostatin-14 (SS-14; 100, 400, and 800 pmol.kg-1.h-1) on plasma levels of SS immunoreactivity (SS-IR) and on pentagastrin-stimulated gastric and pancreatic secretion were compared in conscious dogs. CGRP caused significant inhibition of gastric acid (85-102%), pancreatic protein (63-86%), and pancreatic bicarbonate (74-89%) outputs and a simultaneous dose-related rise (40-102 fmol/ml) in plasma SS-IR. Cessation of CGRP infusion resulted in prompt return of plasma SS-IR to basal levels and an increase in gastric and pancreatic secretion. Although CGRP is a potent releasor of SS into the circulation, its inhibitory action on gastric acid secretion cannot be explained solely by a rise in plasma SS-IR. In the pancreas, in contrast to the stomach, inhibition appears to be more closely related to a rise in circulating level of SS-IR. PMID- 2565090 TI - Neurohumoral changes during onset and offset of ovine heart failure: role of ANP. AB - The temporal relationship of changes in atrial natriuretic peptide (ANP) secretion to other pathophysiological changes in heart failure has not been investigated. We studied the hemodynamic, hormonal, and metabolic changes before, during, and after the induction of heart failure in eight sheep using a 14-day period of rapid left ventricular pacing (LVP). Arterial pressure declined 21% on the first day, while cardiac output fell progressively to 48% of base line, and atrial pressures rose to a plateau over the first week. Plasma ANP levels increased 10-fold with a close correlation to left atrial pressure in all sheep. Furthermore, ANP secretion appeared to be sustained throughout the LVP period but did not prevent avid sodium retention. Marked and early activation of the renin angiotensin system was observed, whereas the major increase in plasma aldosterone commenced 4 days later. On termination of LVP, a prompt natriuresis and diuresis occurred with return of all parameters toward base line. Thus this ovine model is useful for studying pathophysiological changes during the onset and offset of heart failure. PMID- 2565091 TI - Coronary flow during exercise after selective alpha 1- and alpha 2-adrenergic blockade. AB - This study was carried out to determine the relative importance of alpha 1- and alpha 2-adrenergic vasoconstriction in opposing the increase in coronary blood flow, which occurs during exercise. The response of left circumflex coronary artery blood flow was examined during treadmill exercise in 16 chronically instrumented dogs during control conditions, after selective alpha 1-adrenergic blockade with intracoronary prazosin, and after alpha 2-blockade with intracoronary idazoxan. During control conditions, graded treadmill exercise resulted in progressive increases of coronary blood flow and decreases of coronary vascular resistance. Prazosin produced highly selective alpha 1 adrenergic blockade; coronary blood flow was significantly higher and coronary vascular resistance significantly lower during all but the heaviest exercise stage after prazosin. Idazoxan produced highly effective, but only moderately selective, alpha 2-adrenergic blockade. However, after idazoxan, coronary blood flow and coronary vascular resistance during exercise were not significantly different from control. Combined alpha 1- and alpha 2-adrenergic blockade was not more effective in increasing coronary blood flow during exercise than was alpha 1 adrenergic blockade alone. These data support a role for alpha 1-adrenergic coronary vasoconstriction in limiting the increase in coronary blood flow, which occurs during exercise, but do not support a role for alpha 2-mediated coronary vasoconstriction during exercise. PMID- 2565092 TI - Characterization of coronary vasoconstrictor site in medullary reticular formation. AB - The importance of sympathetic neural influences in regulating coronary blood flow has been well established. However, central nervous system pathways responsible for these effects are largely unknown. In a feline model, we have identified a site in medullary reticular formation that may play a role in neural control of the coronary circulation. Changes in heart rate (HR), mean arterial pressure (AP), Doppler coronary flow velocity (CBFV), and femoral flow velocity (FBFV) were measured in 67 anesthetized cats. Electrical stimulation in a specific region of the right medullary lateral reticular formation produced elevations in HR (12 +/- 2% from 156 beats/min), AP (41 +/- 6% from 83 mmHg), CBFV (33 +/- 7%), and femoral vascular resistance index (136 +/- 27%). After beta-adrenergic blockade (propranolol), a transient (5-15 s) stimulus-induced decrease in CBFV was observed in 67% of animals, with a 55 +/- 6% increase in coronary vascular resistance index, not the result of autoregulation. Ipsilateral stellate ganglionectomy or systemic alpha 1-adrenergic blockade abolished the CBFV decrement. Microinjection of L-glutamate into this medullary region failed to elicit either pressor or coronary vasomotor responses. It is concluded that electrical stimulation in a specific site within medullary reticular formation produces neurogenic coronary vasoconstriction as part of a more generalized activation of central sympathetic fibers. This brain stem site may play an important role in reflex or behaviorally mediated coronary responses. PMID- 2565093 TI - Comparison of the effects of famotidine and ranitidine on gastric secretion in patients undergoing elective surgery. AB - A randomised double-blind comparison of oral famotidine and ranitidine given 2 hours before induction, on gastric secretion (volume and pH) was carried out on 93 patients undergoing elective surgery. Gastric contents were aspirated immediately after tracheal intubation. Famotidine significantly reduced the gastric volume, compared with the other groups, including ranitidine. Both famotidine and ranitidine significantly elevated gastric pH towards neutral, compared with the other groups. There was no significant difference between ranitidine and famotidine in respect of the pH. The patients premedicated with famotidine and ranitidine were well protected against Mendelson's syndrome, whereas 38% of patients from the other groups remained at risk. PMID- 2565094 TI - Effect of vecuronium on atropine-induced changes in heart rate. AB - The effect of vecuronium on the heart rate response to atropine has been studied by comparing dose-response relationships in two groups of patients who underwent extracorporeal shock wave lithotripsy. One group received vecuronium (0.1 mg/kg) and the other acted as control. Incremental doses of atropine (1.8, 1.8, 3.6, 7.2 and 14.4 micrograms/kg) were administered and changes in heart rate recorded. No significant differences were observed between the two groups following each incremental dose of atropine. PMID- 2565095 TI - Phenytoin-induced resistance to vecuronium. PMID- 2565096 TI - Does labetalol affect neuromuscular recovery? PMID- 2565097 TI - Effect of terfenadine on cold air-induced bronchospasm. AB - Cold air hyperventilation challenge (CAHC) has been shown to induce bronchoconstriction in asthmatics. We investigated whether terfenadine, a non sedating H1 antihistamine, had a protective effect against CAHC. Twelve mild moderate asthmatics, sensitive to both methacholine and CAHC, underwent a double blind 3-way crossover study of terfenadine at doses of 0, 120, and 240 mg. For four hours after administration of the study drug, pulmonary responses were measured, followed by a CAHC. There was a significant improvement in pulmonary function on the 120-mg and 240-mg days. The percent drop in FEV1 to CAHC was determined by comparing the pre-challenge baseline to the challenge responses. There was a significant attenuation in the FEV1 fall immediately after challenge on active drug days, but it was not sustained. Terfenadine appears to have an attenuating effect against CAHC-induced bronchoconstriction. PMID- 2565098 TI - Comparative study of astemizole and terfenadine in the treatment of chronic idiopathic urticaria. A randomized double-blind study of 40 patients. AB - Forty patients with chronic urticaria were treated with terfenadine or astemizole in a randomized double-blind study. The intensity of the symptoms was quantified and documented daily by the patients. Both antihistamines had a clear effect on the intensity of the pruritus and the severity of the wheals. Neither astemizole nor terfenatine produced any sedative side effects. PMID- 2565099 TI - Quantitative bacterial cultures and beta-lactamase activity in chronic suppurative otitis media. AB - Aspiration of the exudate through open perforation was performed in 54 children with chronic suppurative otitis media. Eighty aerobic and 81 anaerobic isolates were recovered. Aerobic bacteria only were involved in 20 patients (37%), and anaerobic organisms only in seven (13%). Mixed aerobic and anaerobic isolates were recovered from 27 patients (50%). The most common bacteria isolated were anaerobic gram-positive cocci, Bacteroides melaninogenicus group, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Staphylococcus aureus. There were 45 beta lactamase-producing bacteria (BLPB), 30 aerobes, and 15 anaerobes recovered from 38 patients (70%). beta-Lactamase activity was detected in 30 of the 38 ear aspirates (79%) that contained BLPB. All but one of these organisms were in excess of 10(4) colony-forming units/mL. The detection of beta-lactamase activity in the ear aspirates provides evidence of the role of BLPB in the failure of penicillin therapy to eradicate chronic ear infection. PMID- 2565100 TI - [Role of beta-blockers in the treatment of arterial hypertension]. AB - The introduction, more than twenty years ago, of beta-blockers in the treatment of arterial hypertension, represented a significant advance. With these medications, many hypertensive patients are effectively under control and malignant hypertension is practically inexistent. Today, the treatment of hypertension is markedly improved with the development of new, active medications, while the beta-blockers family has markedly evolved. The role of beta-blockers in the treatment of hypertension, must therefore be re-evaluated according to their properties as compared to those of other classes of antihypertensive medications. Indeed, there are standard contraindications to he use of beta-blockers, sometimes resulting in adverse reactions, either clinical (fatigue, sexual disorders, vasomotor syndromes)--much less frequent with the new molecules--or biological (especially serum lipid levels), the consequences of which remains ill-defined--some beta-blockers appear practically without any harmful effect. Actually, despite these drawbacks, usually minimal, there are numerous and strong arguments in favor of the use of beta-blockers in the treatment of hypertension: 1) their significant follow-up in the treatment of hypertension; this is a well-known argument; 2) their effectiveness in hypertension, as a single drug and single daily dose; 3) their cost, which is lower than that of new anti-hypertensive medications; 4) their cardio-protective role, demonstrated by experimental data (myocardial protection and anti arrhythmic effect in experimental ischemia), and clinical data (improvement of left ventricular hypertrophy, control of blood pressure increase during exertion and stress, secondary prevention after myocardial infarction).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565101 TI - Conversion of canavanine to alpha-keto-gamma-guanidinooxybutyrate and to vinylglyoxylate and 2-hydroxyguanidine. AB - It was observed previously that hydroxyguanidine is formed in the reaction of canavanine(2-amino-4-guanidinooxybutanoate) with amino acid oxidases. The present work shows that hydroxyguanidine is formed by a nonenzymatic beta,gamma elimination reaction following enzymatic oxidation at the alpha-C and that the abstraction of the beta-H is general-base catalyzed. The elimination reaction requires the presence in the alpha-position of an anion-stabilizing group--the protonated imino group (iminium ion group) or the carbonyl group. The iminium ion group is more activating than the carbonyl group. Elimination is further facilitated by protonation of the guanidinooxy group. The other product formed in the elimination reaction was identified as vinylglyoxylate (2-oxo-3-butenoate), a very highly electrophilic substance. The product resulting from hydrolysis following oxidation was identified as alpha-keto-gamma-guanidinooxybutyrate (ketocanavanine). The ratio of hydroxyguanidine to ketocanavanine depended upon the concentration and degree of basicity of the basic catalyst and on pH. In the presence of semicarbazide, the elimination reaction was prevented because the imino group in the semicarbazone derivative of ketocanavanine is not significantly protonated. Incubation of canavanine with 5'-deoxypyridoxal also yielded hydroxyguanidine. Since the elimination reactions take place under mild conditions, they may occur in vivo following oxidation at the alpha-C of L canavanine (ingested or formed endogenously) or of other amino acids with a good leaving group in the gamma-position (e.g., S-adenosylmethionine, methionine sulfoximine, homocyst(e)ine, or cysteine-homocysteine mixed disulfide) by an L amino acid oxidase, a transaminase, or a dehydrogenase. Therefore, vinylglyoxylate may be a normal metabolite in mammals which at elevated concentrations may contribute to the in vivo toxicity of canavanine and of some of the other above-mentioned amino acids. PMID- 2565102 TI - [Expression and function of proteins associated with multidrug resistance]. AB - We have identified the proteins specifically expressed in multidrug-resistant tumor cells and studied the functions of these proteins. The 170-to 180-kDa membrane glycoprotein (P-glycoprotein) is an ATPase which works as a pump molecule transporting chemotherapeutic drugs outside the resistant cells. The 22 kDa soluble protein (sorcin) is a calcium binding protein of unknown function. The 85-kDa membrane protein is specifically expressed in adriamycin-resistant cells and induced by treatment with adriamycin, suggesting a mechanism unique for adriamycin resistance. Our monoclonal antibodies to these proteins may well become useful tools for the diagnosis of clinical drug resistance. PMID- 2565103 TI - [Detection of multidrug resistant cells in human malignant diseases by monoclonal antibodies and strategy to eradicate resistant malignant cells]. AB - Two monoclonal antibodies of F (ab')2 form, MRK 16 and MRK 20 that recognize P glycoprotein and P85 kD protein respectively, were useful to detect multidrug resistant cells in human lymphoma, leukemia and gastrointestinal cancer cell lines. They were classified into 4 groups: Group I (4 cell lines) was insensitive to vinca alkaloids, anthracyclines, etoposide (VP-16) and actinomycin-D (ACT-D), and reactive to MRK 16 and MRL 20. Group II (2 cell lines) was insensitive to vincristine (VCR), but not reactive to both antibodies. Group III (3 cell lines) was insensitive to anthracyclines and VP-16, but sensitive to vinca alkaloids and ACT-D, and reactive to MRK 20 but not to MRK 16. Group IV (all other cell lines) was sensitive to these drugs, and not reactive to both antibodies. MRK 16 detects P-glycoprotein-associated multidrug resistance (MDR), while MRK 20 detects P 85kd associated novel MDR. These monoclonal antibodies were useful for detection of MDR cells in clinical samples. PMID- 2565104 TI - [Analysis of VH genes which encode the variable region of monoclonal antibodies directed to cancer-associated carbohydrate antigens]. AB - Immune responses against cancer-associated, carbohydrate antigens are investigated by studying idiotypic determinants of specific antibodies with monoclonal anti-idiotypic antibodies, and by analyzing the structure of VH genes which encode the V region of the anti-carbohydrate antibodies. Four syngenic anti idiotypic antibodies towards monoclonal antibodies which are specific to the sialyl Lewis A antigen and two kinds of SSEA-1 related antigens (sialyl SSEA-1 and fucosul SSEA-1) were obtained. Antibodies directed to carbohydrate antigens were mostly of IgM isotype, indicating these antigens are T-independent antigens, while anti-idiotypic monoclonal antibodies directed to those antibodies were mostly of IgG isotype, suggesting that T cells participate actively in the anti idiotypic response. The Northern blotting analysis of VH genes of monoclonal antibodies directed to negatively-charged carbohydrate antigens such as gangliosides or sulfated glycolipids expressed the VH gene family J558 (group 1), followed by J606 (group 6) and Q52 (group 2) families. On the other hand, monoclonal antibodies directed to SSEA-1 related neutral carbohydrate antigens expressed VH genes of a minor family such as X24 (group 4), V31 (group 9), or 7183 (group 5). The same VH family as expressed in anti-SSEA-1 antibody (x24) was also expressed in the antibodies such as anti-i and anti-I antibodies, which are directed to the synthetic precursors of the SSEA-1 antigen. In either case, the antibodies directed to one particular carbohydrate antigen tended to express the VH gene of one particular family exclusively. This suggests idiotypic homogeneity of the anti-carbohydrate antibodies. PMID- 2565106 TI - The elusive 'cellulite'. PMID- 2565105 TI - Comparative renal effects of intravenous administration of fenoldopam mesylate and sodium nitroprusside in patients with severe hypertension. AB - We studied the renal effects of intravenous administration of fenoldopam mesylate, a dopamine-1 agonist, vs sodium nitroprusside following acute reduction of blood pressure (BP) in 11 patients with severe hypertension (supine BP, 168/124 to 252/135 mm Hg). Following randomization (open-label), timed urinary and plasma samples for clearance of urea and creatinine and excretion of sodium, potassium, and calcium were obtained as well as plasma renin activity for a two hour collection prior to infusion, during a two-hour period of BP control (supine diastolic BP, 95 to 110 mm Hg), and following two hours off the drugs. Mean arterial pressure was lowered similarly with the two drugs (-22% on fenoldopam vs -20% on nitroprusside; P = NS), and neither plasma renin activity nor plasma aldosterone concentration were changed by either drug. However, patients receiving fenoldopam had significant increases in urinary flow and excretion of sodium, potassium, and calcium, whereas patients receiving nitroprusside had no changes in these parameters. Patients receiving fenoldopam had a net fluid balance of -334 mL from the end of baseline to the end of the treatment period, while the nitroprusside group had a positive balance of 382 mL. Thus, these findings show that acute BP reduction with fenoldopam is associated with both a diuresis and natriuresis in severely hypertensive patients while lowering BP with nitroprusside does not predictably alter renal function and causes a moderate expansion in volume. PMID- 2565107 TI - An evaluation of cognitive-behavioral therapy for training resistance to visually induced motion sickness. AB - This investigation examined the techniques for reducing visually-induced motion sickness. On the basis of their responses to a motion sickness history questionnaire, 32 subjects were selected and assigned to 1 of 4 groups such that the groups were matched on the basis of their ability to tolerate visually induced apparent motion (VM). One group received 10 sessions of desensitization training only (DT); a second group received 10 sessions of cognitive therapy only (CT); a third group received 10 sessions of combined desensitization and cognitive therapy treatment (CG); and a fourth group received no treatment (C). (There are many speculations about why and how an individual's response changes with repeated stimulation. We have arbitrarily selected the term desensitization to connote the decrease in sensitivity over time with repeated exposures). The results indicated that only the groups which received cognitive therapy (i.e., CT and CG) exhibited significant increases in tolerance to VM when pretreatment measures were compared to posttreatment measures. No significant differences in pre- vs. posttreatment measures were observed in the desensitization only or control groups (i.e., DT and C). A similar pattern of results was obtained with the symptomatology data. PMID- 2565108 TI - Passenger fear of flying: behavioural treatment with extensive in-vivo exposure and group support. AB - The techniques used in the treatment of passenger fear of flying are described. They include behaviour modification techniques (relaxation, systematic desensitization, and cognitive restructuring), an educational programme, high in vivo exposure, and group support. Results from self-rating scales and qualitative feedback from course participants during 1979-1985, show that the collective use of these techniques contribute significantly to a reduction of fear and discomfort associated with passenger flying. PMID- 2565109 TI - Glutamate uptake into synaptic vesicles of bovine cerebral cortex and electrochemical potential difference of proton across the membrane. AB - Measurements have been made of the ATP-dependent membrane potential (delta psi) and pH gradient (delta pH) across the membranes of the synaptic vesicles purified from bovine cerebral cortex, using the voltage-sensitive dye bis[3-propyl-5 oxoisoxazol-4-yl]pentamethine oxanol and the delta pH-sensitive fluorescent dye 9 aminoacridine respectively. A pre-existing small delta pH (inside acidic) was detected in the synaptic vesicles, but no additional significant contribution by MgATP to delta pH was observed. In contrast, delta psi (inside positive) increased substantially upon addition of MgATP. This ATP-dependent delta psi was reduced by thiocyanate anion (SCN-), a delta psi dissipator, or carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone (FCCP), a protonmotive-force dissipator. Correspondingly, a substantially larger glutamate uptake occurred in the presence of MgATP, which was inhibited by SCN- and FCCP. A nonhydrolysable analogue of ATP, adenosine 5'-[beta gamma-methylene]triphosphate, did not substitute for ATP in either delta psi generation or glutamate uptake. The results support the hypothesis that a H+-pumping ATPase generates a protonmotive force in the synaptic vesicles at the expense of ATP hydrolysis, and the protonmotive force thus formed provides a driving force for the vesicular glutamate uptake. The delta psi generation by ATP hydrolysis was not affected by orthovanadate, ouabain or oligomycin, but was inhibited by N-ethylmaleimide, quercetin, trimethyltin, 7 chloro-4-nitrobenzo-2-oxa-1,3-diazole and 4-acetamido-4'-isothiocyanostilbene 2,2'-disulphonic acid. These results indicate that the H+-pumping ATPase in the synaptic vesicle is similar to that in the chromaffin granule, platelet granule and lysosome. PMID- 2565111 TI - Effect of inhibition of glutathione synthesis on the metabolism and protein conjugation of [14C]captopril in the rat. PMID- 2565110 TI - Stimulation of hepatic cholesterol biosynthesis by fatty acids. Effects of oleate on cytoplasmic acetoacetyl-CoA thiolase, acetoacetyl-CoA synthetase and hydroxymethylglutaryl-CoA synthase. AB - The effects of oleic acid on the activities of cytosolic HMG-CoA (3-hydroxy-3 methylglutaryl-CoA) synthase, AcAc-CoA (acetoacetyl-CoA) thiolase and AcAc-CoA synthetase, as well as microsomal HMG-CoA reductase, all enzymes in the pathway of cholesterol biosynthesis, were studied in the isolated perfused rat liver. Oleic acid bound to bovine serum albumin, or albumin alone, was infused for 4 h at a rate sufficient to sustain an average concentration of 0.61 +/- 0.05 mM fatty acid during the perfusion. Hepatic cytosol and microsomal fractions were isolated at the termination of the perfusion. Oleic acid simultaneously increased the activities of the cytosolic cholesterol-biosynthetic enzymes 1.4-2.7-fold in livers from normal fed rats and from animals fasted for 24 h. These effects were accompanied by increased net secretion by the liver of cholesterol and triacylglycerol in the very-low-density lipoprotein (VLDL). We confirmed the observations reported previously from this laboratory of the stimulation by oleic acid of microsomal HMG-CoA reductase. In cytosols from perfused livers, the increase in AcAc-CoA thiolase activity was characterized by an increase in Vmax. without any change in the apparent Km of the enzyme for AcAc-CoA. In contrast, oleic acid decreased the Km of HMG-CoA synthase for Ac-CoA, without alteration of the Vmax. of the enzyme. The Vmax. of AcAc-CoA synthetase was increased by oleic acid, and there was a trend towards a small increase in the Km of the enzyme for acetoacetate. These data allow us to conclude that the enzymes that supply the HMG-CoA required for hepatic cholesterogenesis are stimulated, as is HMG-CoA reductase, by a physiological substrate, fatty acid, that increases rates of hepatic cholesterol synthesis and cholesterol secretion. Furthermore, we suggest that these effects of fatty acid on hepatic cholesterol metabolism result from stimulation of secretion of triacylglycerol in the VLDL by fatty acids, and the absolute requirement of cholesterol as an important structural surface component of the VLDL necessary for transport of triacylglycerol from the liver. PMID- 2565113 TI - Genetic analysis of CR1 expression on erythrocytes of patients with systemic lupus erythematosus. AB - The role of genetic factors in decreased expression of CR1 in patients with systemic lupus erythematosus (SLE) was investigated by assessing the frequency of genotypes determining the numbers of CR1 on erythrocytes obtained from 52 patients with SLE and from 84 normal individuals. The expression of CR1 was quantitated using flow cytometry. Genotypes were determined by analyzing the CR1 gene restriction fragment length polymorphism using the CR1.1 complementary DNA probe and the Hind III restriction enzyme. In normal subjects, the distribution of the HH, HL, and LL genotypes fit the Hardy-Weinberg law. The frequency for the H allele did not differ between the 2 groups. No individual homozygous for the LL genotypes was found among the SLE patient population. Taken together, data from this and previous studies indicate an under-representation of the LL homozygous genotype in patients with SLE. SLE patients expressed decreased numbers of CR1 per erythrocyte within each genotype compared with these numbers in normal subjects. The results suggest that defective expression of CR1 in SLE patients is acquired and that the presence of the L allele is not linked to a genetic susceptibility for SLE. PMID- 2565112 TI - Drug metabolism by periportal and perivenous rat hepatocytes. Comparison of phase I and phase II reactions and their inducibility during culture. AB - Hepatocytes were aseptically isolated from either the periportal (pp; zone 1) or the perivenous (pv; zone 3) region by digitonin-collagenase perfusion and cultured on type I collagen for 4 to 9 days. In freshly isolated cells the pp:pv activity ratios of the acinar marker enzymes gamma-glutamyltranspeptidase (gamma GT), alanine aminotransferase (ALAT) and glutamate dehydrogenase (GLDH) were 2.8, 1.6 and 0.76, respectively. During culture ALAT and GLDH activities gradually declined, but the pp-pv difference was retained for at least 4 days. In contrast, the difference in the gamma-GT activity was rapidly lost, due to its fast initial activation in pv cells. The initial 7-ethoxycoumarin O-deethylase (ECDE) activity was higher in pv cells; this difference was retained for several days of culture and was increased by induction in vitro with either phenobarbital (PB) or beta naphthoflavone (beta NF). Although the basal UDP-glucuronyltransferase (UDPGT) activity with either p-nitrophenol (pNP) or hydroxybiphenyl (HBP) as substrate did not differ significantly, the in-vitro PB- or beta NF-induced activity was higher in pv cells. Both glucuronidation and sulfation of methylumbelliferone tended to be higher in pv cells. Glutathione S-transferase was initially significantly higher in pv cells and this difference was augmented after in vitro induction by PB or beta NF. After six days in culture all the observed pp-pv differences had disappeared. These results suggest that hepatocytes isolated from the perivenous region seem to maintain their initially higher capacity for phase I and phase II drug reactions during culture and also respond more strongly than periportal cells to in vitro induction. PMID- 2565114 TI - n-3 fatty acid deficiency in man. AB - A total of nine patients with n-3 fatty acid deficiency are described. They had been fed by gastric tube for 2.5-12 years, and had received 0.02-0.09% of calories as n-3 acids. The observed clinical symptoms of n-3 fatty acid deficiency were scaly and haemorrhagic dermatitis, haemorrhagic folliculitis of the scalp, growth retardation and impaired wound healing. All patients had extensive brain damage making it impossible to evaluate n-3 effects on cerebral functions. The patients were supplemented with cod liver and soya oil, alpha linolenate followed by a purified fish oil, or with a mixture of linseed and cod liver oil. The results indicate that a dietary supply of 1.0-1.2% of alpha linolenic acid is necessary to obtain a mid-normal lipid concentration of n-3 fatty acids, and suggest that the minimal dietary requirement is 0.2-0.3% of total calories. Long-chain n-3 fatty acids seem to be approximately twice as efficient as alpha-linolenic acid in maintaining normal n-3 fatty acid concentrations in plasma and red cell lipids. PMID- 2565115 TI - Interactions between the metabolism of n-3 and n-6 fatty acids. AB - Dietary n-3 fatty acids modify the fatty acid composition of phospholipids of different cells and tissues from rats in diverse ways. Neutrophil and platelet phospholipids contain elevated amounts of 20:5n-3, but only relatively small changes occur in the levels of 22-carbon n-3 fatty acids. Conversely, dietary n-3 acids result primarily in an increase in 22-carbon n-3 acids in heart, liver and kidney phospholipids. Platelets metabolize exogenous n-6 and n-3 fatty acids into a variety of different autocoids. However, it appears that only arachidonate and 20:5n-3 are released from phospholipids upon agonist-induced stimulation of phospholipases. Neutrophils metabolize arachidonate and 20:5n-3 in similar ways, both relative to phospholipid biosynthesis and the subsequent release of these acids for metabolism into leukotrienes. PMID- 2565116 TI - Pro-Leu-Gly-NH2 serves as a conditioned stimulus in the acquisition of conditioned tolerance. AB - The effect of Pro-Leu-Gly-NH2 (MIF) on the acquisition of tolerance to morphine induced antinociception and its efficacy as a cue predictive of morphine administration was examined. Daily administration of MIF prior to morphine injection did not attenuate the acquisition of tolerance to the antinociceptive properties of morphine, as measured by the latency to hindpaw lick in a hot-plate test of analgesia. When the animals were tested 72 hr later without MIF pretreatment, they appeared to lose tolerance, as indicated by longer latencies to paw lick. These data suggest that in some situations MIF may interfere with the acquisition of tolerance by acting as a cue that reliably predicts the antinociceptive properties of morphine. PMID- 2565117 TI - Cardiovascular responses to metipranolol and timolol eyedrops in healthy volunteers. AB - 1. Intraocular pressure and cardiovascular responses to metipranolol 0.1% and 0.3% and timolol 0.25% eyedrops were measured in a balanced single dose placebo controlled crossover study in eight healthy volunteers aged 34-58 years. 2. Timolol 0.25% and metipranolol 0.3% reduced intraocular pressure throughout the 6 h period of observation to a similar extent. Metipranolol 0.1% was marginally less effective, significantly reducing pressure up to 4 h only. 3. No drug treatment significantly altered resting heart rate or blood pressure. Timolol 0.25% significantly reduced exercise tachycardia (P less than 0.05), an effect which was not shown by metipranolol 0.1 or 0.3%. Exertional pain in the legs occurred more frequently after timolol 0.25% and metipranolol 0.3% than after metipranolol 0.1% or placebo eyedrops. 4. Octan-1-ol/pH 7.4 buffer distribution coefficients at 37 degrees C were found to be: metipranolol 5.19, timolol 0.84, indicating that metipranolol has an approximately 6-fold greater lipid solubility. 5. It is concluded that, by comparison with timolol, metipranolol in eyedrop concentrations up to 0.3%, despite its greater lipid solubility, reaches concentrations in the systemic circulation which are less likely to affect the heart. PMID- 2565118 TI - Beta-adrenoceptors and human skeletal muscle characterisation of receptor subtype and effect of age. AB - 1. Rectus abdominis muscle biopsies were obtained from 28 patients undergoing abdominal surgery. In membranes prepared from these biopsies beta-adrenoceptor binding was examined. The apparent affinity (KD) and the density (Bmax) of the receptors for the radioligand (-)-[125I]cyanopindolol were 28.5 +/- 2.7 (pM) and 25.9 +/- 2.1 (fmol mg-1 protein) (mean +/- s.e. mean) respectively. In forceps biopsies from vastus lateralis muscle from four healthy volunteers the values for KD and Bmax were 22.5 +/- 4.4 (pM) and 16.4 +/- 2.2 (fmol mg-1 protein). The binding characteristics for the radioligand were similar in the biopsies from the two muscle sites. 2. Inhibition of the radioligand binding by the selective beta 2-adrenoceptor antagonist ICI 118551 (KI = 117 +/- 45 nM) and selective beta 1 adrenoceptor antagonist metoprolol (KI = 15229 +/- 5046 nM) suggests the dominance of beta 2-adrenoceptor subtype in human skeletal muscle. 3. There were no significant differences in the skeletal muscle beta-adrenoceptor densities or affinities between the young and older patients. PMID- 2565119 TI - A comparison of the influence of famotidine and cimetidine on phenytoin elimination and hepatic blood flow. AB - The H2-receptor antagonist cimetidine has been reported to decrease the hepatic clearance of numerous drugs by inhibiting cytochrome P-450 metabolism, decreasing liver blood flow or both. In this open-label, randomized crossover study we determined whether therapeutic doses of famotidine, a newer H2-receptor antagonist, has similar effects. Ten healthy subjects received single doses of both phenytoin 100 mg orally and indocyanine green intravenously without other treatment, and then again during treatment with famotidine or cimetidine. After a drug-free period, this sequence was repeated with the alternate H2-receptor antagonist. Cimetidine decreased the plasma clearance of phenytoin by 16% +/- 14% (mean +/- s.d.), but was not found to have a significant influence on phenytoin volume of distribution or terminal elimination rate constant nor on blood clearance of indocyanine green. Famotidine was not found to alter either phenytoin or indocyanine green kinetics. PMID- 2565120 TI - Molecular basis and population genetics of phenylketonuria. PMID- 2565121 TI - Effect of the beta-agonist cimaterol on growth and composition of neonatal rats selected for large and small body size. AB - The effect of subcutaneous injection of the beta-agonist cimaterol on growth and body composition of neonatal rats differing in growth potential was examined. Rats that represented substrain populations of Charles River CD rats selected for either large or small body size were used. Cimaterol administration resulted in a greater reduction in body weight gain in the Large strain rats. Body growth rate declined linearly with increasing doses of cimaterol in both strains. Percent carcass fat and protein were unchanged with cimaterol treatment. Administration of cimaterol inhibited skeletal muscle growth but caused cardiac hypertrophy. These results suggest that the effectiveness of beta-agonists is influenced by animal genotype. PMID- 2565122 TI - Mode of bilirubin deposition in the cerebellum of developing jaundiced Gunn rats. AB - Bucolome is known as a potent displacer of bilirubin from the blood into tissues. The effects of the drug on newborn rats with congenital jaundice (Gunn rats) were examined on postnatal days 7, 11, 15 and 21. A single subcutaneous injection of bucolome resulted in a rapid fall of the total plasma bilirubin concentration and the lowered level persisted as long as 23 h. Concomitantly with the drop, the cerebellar bilirubin level increased within 1 h. Although cerebellar bilirubin returned to the initial level after 24 h in rats treated with bucolome on days 7 and 21, it remained almost constant or rather increased during the period of 1-24 h in rats treated on days 11 or 15. In 15-day-old rats, localized yellow staining in the cerebellum 24 h after treatment was most apparent in the granule cell layer of the ventral part of the pyramis and the dorsal part of the uvula under the dissecting microscope. These observations suggest that the period most susceptible to bilirubin deposition lies around day 15 in the cerebellum of jaundiced Gunn rats. PMID- 2565124 TI - Pain control in breast cancer. A panel discussion. AB - Pain can be a prominent finding in breast cancer patients. It may occur in the setting of the postmastectomy period, related to the disruption of normal neural pathways or the development of lymphedema. In advanced disease, the management of pain from nerve compression or bone metastases requires special approaches. In this panel discussion, the participating physicians will discuss these topics and provide an up-to-date approach to pain control in breast cancer patients. PMID- 2565123 TI - Detection of P-glycoprotein in human leukemias using monoclonal antibodies. AB - Overexpression of a Mr 170,000 membrane glycoprotein (P-glycoprotein) is consistently associated with multidrug resistance in cell lines. Two monoclonal antibodies (Mab) against P-glycoprotein (265/F4 and C 219) were used to examine tumour samples from patients with leukemias for evidence of P-glycoprotein overexpression. High levels of P-glycoprotein (greater than 5% positive cells) were detected with both antibodies in samples from 3 out of 18 patients suggesting that a multidrug resistant phenotype may also occur in human leukemias. PMID- 2565125 TI - Applications of modified electrodes in the voltammetric determination of catecholamine neurotransmitters. PMID- 2565126 TI - The International Collegium of Rehabilitative Audiology. PMID- 2565127 TI - Genotypic analysis of engraftment in thalassemia following bone marrow transplantation using synthetic oligonucleotides. AB - DNA hybridization with synthetic oligonucleotide probes was used to assess engraftment in 19 thalassemic patients who received bone marrow grafts from their respective healthy HLA-identical siblings. Three oligomers complementary to the tandem repetitive sequences of different hypervariable regions of human DNA were designed so as to produce simple RFLP (restriction fragment length polymorphism) patterns. Each probe hybridizes to one or two bands in HinfI-digested genomic DNA. The combined use of these three probes allowed a discrimination between all the HLA-identical siblings tested. Both donor-specific and recipient-specific DNA fragments existed in 18 out of the 19 sibling pairs studied. One pair possessed only a donor-specific fragment. DNA analysis at an early stage after the graft detected donor-specific fragments in 15 out of 19 patients, recipient-specific fragments in three patients and a mix of recipient and donor fragments in one patient. At a later stage this patient possessed donor-specific fragments only. Follow-up DNA analysis confirmed these findings. Thus 16 patients continued to display donor-specific fragments over 60 days post-transplant. These DNA data showed strong correlation with the clinical status of the patients as well as with other markers of engraftment including cytogenetics and hemoglobin synthesis. The patients who showed donor-specific fragments over 60 days have been free of thalassemic symptoms for over 300 days. Moreover, in 11 cases it was possible to predict the fate of the graft within 15 days after transplantation. In conclusion, the use of the three synthetic oligonucleotide probes provides a powerful tool in documenting engraftment in bone marrow transplantation. PMID- 2565128 TI - Sulphasalazine in rheumatoid arthritis: haematological problems and changes in haematological indices associated with therapy. AB - This prospective study documents the haematological responses in 300 rheumatoid patients (RA) treated with sulphasalazine (SASP) for between 1 and 9 years. It also examines the effect of SASP on the total white cell and platelet counts over 2 years in relation to disease activity in 80 RA patients. Neutropenia occurred in six (2%) (three severe--neutrophil count less than 0.8 X 10(9)/l) after 3 and 12 weeks. The drug was withdrawn in six immediately and in one patient after 21 months when the neutrophil count fell to 0.7 X 10(9)/l. An additional 11 (3.7%) developed mild or transient leucopenia between 2 weeks and 24 months, and eight continued therapy. Thrombocytopenia occurred in one patient at 18 weeks associated with other reactions. Four with Felty's syndrome developed a further fall in the total WBC associated with thrombocytopenia in two. A rise in mean cell volume was common (72%), and macrocytosis (MCV greater than 98 fl) occurred in 27 (9%). Macrocytic anaemia was rare (less than 1%). All haematological problems were reversible. In 80 patients treated with SASP for 2 years there was a significant fall in the median white cell and platelet counts at 3 months associated with improvement in disease activity. PMID- 2565129 TI - Berger's disease with polyarteritis nodosa. AB - We describe a patient with Berger's disease and polyarteritis nodosa. This association has not been described previously in the literature. A causal relationship between the two diseases is suggested. PMID- 2565130 TI - Ten years of sulphasalazine use in rheumatoid arthritis. PMID- 2565131 TI - Effects of a spider toxin (JSTX) on hippocampal CA1 neurons in vitro. AB - The effect of a toxin (JSTX) obtained from Nephila clavata (Joro spider) on the CA1 pyramidal neurons of the hippocampus was studied using slice preparations. JSTX blocked the excitatory postsynaptic potentials (EPSPs) in the pyramidal neuron evoked by Schaffer collateral stimulation but was without effect on the antidromic action potentials or on the resting conductance. Depolarization induced by ionophoretic application of glutamate was readily suppressed by JSTX but aspartate-induced depolarization was much less sensitive to the toxin. Among preferential agonists activating 3 receptor subtypes for excitatory amino acids, quisqualate responses were most effectively suppressed by JSTX. Kainate responses were similarly suppressed but in some cells higher concentration of the toxin was needed to block the responses. N-methyl-D-aspartate (NMDA) responses were the least sensitive to JSTX but they were suppressed by +/- 2-amino-5 phosphonovaleric acid (APV). Long term potentiation (LTP) once it had taken place was not completely inhibited by APV. In the presence of JSTX, however, LTP was blocked and tetanic stimuli produced only a short-lived potentiation. In Mg2+ free solution, an orthodromic stimulation evoked repetitive spike responses which were superimposed on the depolarization following the initial spike. APV suppressed the depolarization and associated spikes leaving an orthodromic response which was sensitive to JSTX. The results suggest that JSTX blocks EPSPs in CA1 pyramidal neurons which are mediated by non-NMDA type receptors. PMID- 2565132 TI - Monoclonal antibody directed against glutaraldehyde conjugated glutamate and immunocytochemical applications in the rat brain. AB - Like other small-sized neurotransmitter molecules, glutamate (Glu) was conjugated to carrier proteins via glutaraldehyde (G). Human serum albumin (HSA) and thyroglobulin (TH) conjugates were alternately injected into mice. When a relevant immune response was obtained for antibody affinity and specificity, hybridization of spleen activated lymphocytes with SP2/O/Ag myeloma cells was performed. Supernatant culture media of hybridomas were tested for the presence of anti-conjugated Glu antibodies with our ELISA method. Selected hybridomas giving good antibody affinity and specificity were then cloned by the limiting dilution technique. Using DEAE-chromatographed ascites fluid, Glu reactivity was observed on the cortex and the hippocampus. Staining obtained with this monoclonal antibody was in agreement with that observed with previous polyclonal antisera directed against conjugated Glu or monoclonal anti-gamma-glutamyl-Glu antibody. PMID- 2565133 TI - Adrenergic mediated phosphatidylinositol metabolism is modulated by epileptic discharges in human neocortex. AB - Adrenergic mediated phosphatidylinositol (PI) hydrolysis was measured in tissues obtained from 21 patients immediately following surgery for focal epilepsy. Accumulation of [3H]inositol monophosphate (IP1) was significantly reduced (21%, P less than 0.01) in actively spiking neocortex (n = 15) versus samples from non spiking regions (n = 9). Epileptic discharges may blunt alpha 1-adrenoceptor stimulated transmembrane signalling in human neocortical epileptic foci. PMID- 2565134 TI - Electron microscopic immunocytochemical evidence for the existence of bidirectional synaptic connections between growth hormone-releasing hormone- and somatostatin-containing neurons in the hypothalamus of the rat. AB - Synaptic contacts between growth hormone-releasing hormone (GHRH)- and somatostatin-containing neurons were demonstrated in the rat hypothalamus by a double-staining immunocytochemical method at the electron microscopic level. Somatostatin-immunoreactive nerve terminals synapse on GHRH-positive dendrites and cell bodies in the arcuate nucleus. A fine network of GHRH-immunopositive nerve terminals was observed at the light microscopic level in the rostral part of the periventricular nucleus and in the dorsal part of the arcuate nucleus around somatostatin-containing neuronal elements. With the electron microscope synaptic contact between GHRH-containing nerve terminals and somatostatin containing dendrites are demonstrated. The reciprocal innervation between GHRH- and somatostatin-containing neurons that project to the median eminence and regulate growth hormone secretion must allow them to coordinate their activities. PMID- 2565135 TI - L-glutamate binding sites of normal and atrophic human cerebellum. AB - The binding kinetics, pharmacologic properties, ontogeny and localization of L glutamate binding sites were studied in membrane preparations and sections of normal and olivopontocerebellar atrophy (OPCA) human cerebellum. One binding component was found with a Kd value in the order of 150 x 10(-9) M. No significant changes of Kd values were observed with age, whereas the highest Bmax value was observed at the age of 1 year. L-Aspartate, ibotenate, quisqualate and L-homocysteic acid were potent inhibitors of L-[3H]glutamate binding. Quantitative densitometric measurements indicated the presence of L-glutamate sites in both the molecular and granule cell layer. In OPCA cerebella a very significant decrease of L-[3H]glutamate specific binding (Bmax) was observed, whereas Kd values were found unchanged. The pharmacologic properties of L [3H]glutamate binding sites of OPCA cerebellar tissues were similar to those of normal cerebellum. [3H]quinuclidinyl benzylate binding, expressed in fmol/mg protein, did not show significant differences between normal and OPCA cerebella. PMID- 2565136 TI - Cardiac responses to the microinjections of excitatory amino acids into the intermediolateral cell column of the rat spinal cord. AB - Sympathoexcitatory cardiovascular responses to the microinjections of L-glutamate into the intermediolateral cell column (IML) of the upper thoracic cord (C8 to T4) were studied. Mean arterial pressure (MAP), heart rate (HR), the rate of increase in the left ventricular pressure (dP/dt) and contractility index were monitored in immobilized and artificially ventilated male Wistar rats anesthetized with pentobarbital or isoflurane. On the right side, microinjections (10-20 nl) of L-glutamate (0.9-1.77 nmol in 0.9% sodium chloride solution, pH 7.4) into the IML at T2 level produced marked tachycardiac responses with relatively small changes in contractility. On the left side, similar microinjections produced marked increase in dP/dt and contractility index with relatively small increase in HR. On either side, the responses were smaller at T1 and T3 level and absent at C8 and T4 level. No changes in blood pressure were observed with microinjections of L-glutamate on either side. Microinjections of N methyl-D-aspartic acid (NMDA), 1-100 pmol, into the IML elicited responses similar to those of L-glutamate. These amino acids failed to evoke any response when microinjected into the adjacent areas (e.g. 0.5 mm lateral or medial to the IML). The effects of glutamate and NMDA in the IML were blocked by microinjections of glutamic acid diethylester (GDEE) and D-2-amino-7 phosphonoheptanoic acid (D-AP7), respectively. Control microinjections of physiological saline into the IML produced no responses. These results indicate that excitatory amino acids, in small doses and volumes, can be used to identify cardiac sympathoexcitatory neuronal pools in the IML. This preparation may prove useful in characterizing pharmacological actions of various putative neurotransmitters in this region of the spinal cord. PMID- 2565137 TI - Cytosolic free calcium and NAD/NADH redox state in the cat cortex during in vivo activation of NMDA receptors. AB - Activation of the N-methyl-D-aspartate (NMDA) receptors and the concomitant Ca2+ entry have been implicated in neuronal injury in a variety of pathological states. The effects of extracellular Mg2+ concentrations and D,L-2-amino-5 phosphonovaleric acid (APV), a competitive NMDA receptor antagonist on the NMDA induced responses were investigated in vivo. In vivo fluorometric measurements were made of changes in cytosolic free Ca2+ ([Ca2+]i) and NADH fluorescence directly from the cat cortex using indo-1, a fluorescent Ca2+ indicator. Changes in [Ca2+]i were assessed utilizing the ratio of indo-1 emission at two wavelengths (400 and 506 nm) during excitation with ultraviolet light (340 nm). Application of 100 microM NMDA to the cortex produced a significant increase in the [Ca2+]i signal ratio at physiological concentrations of Mg2+ (1.2 mM). This increase was enhanced in the absence of Mg2+ and was completely blocked either at 5 mM Mg2+ or in the presence of 50 microM APV. The NAD/NADH redox state was initially oxidized, which was also blocked by either high Mg2+ or APV. The application of NMDA elicited characteristic electroencephalogram (EEG) changes consisting of a marked reduction in amplitude and regular spikes (17-20 Hz). These EEG changes did not appear in the presence of APV. In addition to NMDA receptor antagonists, the level of extracellular Mg2+ is a potent physiological modulator of the NMDA response. PMID- 2565138 TI - Excitatory amino acid-evoked release of [3H]GABA from hippocampal neurons in primary culture. AB - We investigated the release of gamma-[2,3-3H(N)]aminobutyric acid ([3H]GABA) from hippocampal neurons in primary cell culture. [3H]GABA release was stimulated by the excitatory amino acid neurotransmitter glutamate as well as by N-methyl-D aspartate (NMDA) and kainate. Cell depolarization induced by raising [K+]o or by veratridine also stimulated [3H]GABA release. NMDA-induced release was completely blocked by 3-((+/-)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP+), Mg2+ and Zn2+ whereas the release induced by glutamate and kainate was much less susceptible to inhibition by these substances. Furthermore, removal of external Ca2+ inhibited NMDA-induced release, but not that induced by glutamate, kainate, veratridine or 50 mM K+. Removal of external Na+ reduced [3H]GABA release evoked by all stimuli, but to different extents. All of the excitatory amino acids tested increased [Ca2+]i within hippocampal neurons as assessed by fura-2 based microspectrofluorimetry. This increase in [Ca2+]i was completely dependent on the presence of external Ca2+. These results suggest that Ca2+-dependent and independent forms of GABA release from hippocampal interneurons may occur. [3H]GABA release evoked by glutamate, kainate, veratridine or 50 mM K+, appeared to be mediated by the reversal of electrogenic, Na+-coupled GABA uptake. Release was inhibited by nipecotic acid, an inhibitor of the Na+-coupled GABA uptake system. However, release induced by NMDA may also include a Ca2+-dependent component. PMID- 2565139 TI - Cysteamine-induced depletion of somatostatin produces differential cognitive deficits in rats. AB - The effects of a variety of doses of systemically administered cysteamine (a somatostatin depletor) were studied on step-through passive avoidance retention, as well as acquisition and performance of a delayed spatial alternation task and a signaled extinction discrimination task in rats. Retention of single trial passive avoidance was significantly reduced by a pretraining (60-min) dose of cysteamine at 50, 100, 150 and 200 mg/kg s.c. This effect was shown to be sensitive to behavioral manipulation; in a second experiment, a retention deficit was found only at the two highest doses tested (150 and 200 mg/kg s.c.) after a second exposure to the footshock. In the operant conditioning studies, biweekly injections (Monday and Wednesday) of cysteamine administered one hour before testing produced no statistically significant changes in acquisition or performance of either the delayed spatial alternation or the signaled discrimination task. The results of these series of experiments suggest that active somatostatin release or chronic somatostatin depletion may selectively affect performance maintained by different behavioral procedures. PMID- 2565140 TI - Release of endogenous N-acetylaspartylglutamate (NAAG) and uptake of [3H]NAAG in guinea pig cerebellar slices. AB - For the purpose of obtaining chemical information about the physiological role of N-acetylaspartylglutamate (NAAG), the release of endogenous NAAG from and the uptake of [3H]NAAG by Guinea pig cerebellar slices were investigated in comparison with L-aspartate (Asp) and L-glutamate (Glu). Although endogenous NAAG was found to be released spontaneously from the slices as is endogenous Asp and Glu, high-K+-induced facilitation of release occurred only for endogenous Asp and Glu in a Ca2+-dependent manner, but not for NAAG. It was confirmed that [3H]NAAG itself was taken up in a Na+-dependent manner by the slices by two low-affinity processes with small Vmax values, and labeled Glu and glutamine were detected as the metabolites of [3H]NAAG in the slices. The [3H]NAAG uptake was slower than that of labeled Glu and was significantly depressed by NAAG, Asp, Glu and D aspartate, but not affected by gamma-aminobutyrate, suggesting that NAAG may share a common uptake carrier with excitatory amino acids. These results suggest that endogenous NAAG may act extracellularly, but the amount of endogenous NAAG released from nerve terminals by presynaptic depolarization may be very small if any, and also that spontaneously liberated NAAG can be inactivated by low affinity uptake systems, at least, in the Guinea pig cerebellum. PMID- 2565141 TI - Behavioral and biochemical evidence for a functional role of excitatory amino acids in the median raphe nucleus. AB - Recent experiments have suggested the existence of excitatory amino acid (EAA) containing afferents to the median raphe nucleus. In the present study we investigated the functional significance of EAAs in the median raphe (MR) by examining the behavioral and biochemical effects of intra-raphe injections of EAA antagonists. Injections of kynurenic acid, gamma-glutamylglycine, 2-amino-5 phosphonovaleric acid (2-APV) and 2-amino-7-phosphonoheptanoic acid (2-APH) into the median raphe resulted in marked hyperactivity. In contrast to the effect of 2 APV and 2-APH, intra-raphe injections of the homologues of these compounds with 4, 6 or 8 carbon atoms, which have a lower affinity for excitatory amino acid receptors, were without significant effects on activity. Additionally, the effects of 2-APV were shown to be stereospecific to the active D-isomer further suggesting receptor mediation of the effect. Injections of EAA antagonists into the dorsal raphe nucleus or the ventral tegmental area were much less effective in increasing activity than were injections into the MR, suggesting anatomical specificity of the effect. Injections of 2-APV into the median raphe were also shown to result in a reduction of serotonin metabolism within the hippocampus and an increase in dopamine metabolism within the nucleus accumbens and the magnitude of both of these effects was positively correlated with the behavioral responses to the injections. These findings suggest that cells within the median raphe may be subject to a tonic excitation exerted through EAA receptors. PMID- 2565142 TI - NMDA receptor antagonists block norepinephrine-induced long-lasting potentiation and long-term potentiation in rat dentate gyrus. AB - In the in vitro rat dentate gyrus, norepinephrine-induced long-lasting potentiation (NELLP) and long-term potentiation (LTP) of responses to perforant path stimulation were blocked by the N-methyl-D-aspartate (NMDA) receptor antagonists, D(-)-2-amino-5-phosphonovaleric acid (D(-)APV) and 3-[(+/-)-2 carboxypiperazin-4-yl]propyl-1-phosphonic acid (CPP). CPP and D(-)APV, but not L(+)APV, also depressed the orthodromic population spike but not the antidromic spike, which suggests that these receptors may function in low-frequency evoked activity of granule cells. We conclude that NELLP, like LTP in the dentate gyrus, requires NMDA receptor activation. PMID- 2565143 TI - N-methyl-D-aspartate receptor autoradiography in rat brain after angular bundle kindling. AB - The specific binding of L-[3H]glutamate to N-methyl-D-aspartate (NMDA) receptors in brain regions of kindled rats was visualized autoradiographically and quantitated. When assayed 28 days after the last evoked seizure, NMDA receptor binding had declined by 7-11% in stratum radiatum of the dorsal hippocampal area CA1, in both deep and superficial layers of the motor cortex and in layers I-IV of the somatosensory cortex. No significant changes were detected in any other brain region nor in any region examined 1 day after the last evoked seizure. These findings suggest that the enhanced activation of NMDA receptors in kindled rats cannot be explained by an increased expression of these receptors. Rather, kindling leads to a regionally-selective down regulation of NMDA receptor binding. PMID- 2565144 TI - Evidence for a distinct nigropallidal dopaminergic projection in the squirrel monkey. AB - Injections of the retrograde fluorescent tracer fast blue in the striatum (STR) and nuclear yellow in the internal segment of the globus pallidus (GPi) in the squirrel monkey (Saimiri sciureus) revealed a nigropallidal projection whose cellular origin was largely distinct from that of the nigrostriatal pathway. Neurons containing the tracer injected in GPi were scattered throughout the substantia nigra-ventral tegmental area complex where they formed approximately 20-25% of the total number of retrogradely labeled cells. Only about 5-10% of all positive neurons were double-labeled after STR-GPi injections. In experiments combining the use of the fluorescent tracer propidium iodide with immunofluorescence, the majority of neurons projecting to GPi displayed tyrosine hydroxylase immunoreactivity. Hence, in addition to their important role at striatal level, midbrain dopaminergic neurons may influence directly the output neurons of the basal ganglia at pallidal level in primates. PMID- 2565145 TI - Effects of inhibitory and excitatory amino acid neurotransmitters on isolated cerebral parenchymal arterioles. AB - The vasoactive properties of inhibitory (gamma-aminobutyric acid (GABA), glycine) and excitatory (glutamate, aspartate) amino acid neurotransmitters were studied in isolated rat cerebral parenchymal arterioles. None of these neurotransmitters had a significant effect on vessel diameter at concentrations between 10(-9) and 10(-3) M, except for 10(-3) M glycine. These amino acid neurotransmitters are unlikely to play a role in regulation or pathophysiology of the cerebral microcirculation by directly changing vascular diameter. PMID- 2565146 TI - Repeated reserpine administration up-regulates the transduction mechanisms of D1 receptors without changing the density of [3H]SCH 23390 binding. AB - Behavioural studies have shown that stimulation of D1 receptors, which is uneffective in normal rats, induced strong hypermotility in rats pretreated with reserpine for 5 days. On this basis, we investigated D1 receptor plasticity using the 5-day treatment with reserpine (1 mg/kg; s.c.) as an experimental model. The function of striatal D1 receptors was determined both in binding studies with [3H]SCH 23390 and by measuring formation of cAMP in response to the selective agonist, SKF 82526. The results indicate that the responsiveness of adenylate cyclase (AC) to D1 receptor stimulation was markedly increased after reserpine administration, while no significant changes were found in [3H]SCH 23390 binding site density. Moreover, formation of cAMP after stimulation of Gs protein with GppNHp was markedly enhanced in dopamine (DA)-depleted rats; the responsiveness of AC to forskolin, which directly stimulates the AC catalytic unit, was not affected by reserpine administration. These data indicate that reserpine-induced D1 receptor up-regulation is apparently mediated by a marked enhancement of the coupling efficiency of Gs protein, suggesting that the D1 behavioral supersensitivity does not correlate with the density of D1 receptors, but is reflected by a selective up-regulation of their transduction mechanisms. PMID- 2565147 TI - Selective staining of a population of parvalbumin-containing GABAergic neurons in the rat cerebral cortex by lectins with specific affinity for terminal N acetylgalactosamine. AB - Lectins with specific affinity for terminal N-acetylgalactosamine, Vicia villosa agglutinin (VVA) and Glycine max agglutinin (soybean agglutinin; SBA), are shown to stain selectively a subpopulation of GABAergic neurons in the rat cerebral cortex. About 90% of VAA- and/or SBA-stained cells are also parvalbumin immunoreactive, but no VVA-stained cells showed somatostatin-28-like or cholecystokinin-8-like immunoreactivities. PMID- 2565148 TI - Different developmental potentials of diencephalic and mesencephalic dopaminergic neurons in vitro. AB - Morphological and functional differentiation of dopamine (DA) neurons was compared in dissociated cultures from gestational day 14 rat mesencephalon and diencephalon. Numbers of tyrosine hydroxylase-immunoreactive (TH-IR) neurons relative to all neurons were 4 and 1.7 times higher in mesencephalic than in diencephalic cultures at 6 and 13 days in vitro (DIV), respectively. Morphological maturation of diencephalic DA neurons was retarded in comparison to mesencephalic DA neurons. There were gross differences in soma size and length of processes between the two types of DA neurons, the appearance of which was strongly reminiscent of DA cell types described in vivo. Functional maturation of DA neurons was quantified by measuring uptake and Ca2+-dependent K+-stimulated release of [3H]DA per TH-IR neuron. As early as 6 DIV, DA uptake by mesencephalic DA neurons was saturable, was sensitive to benztropine and reserpine, and could be displaced by unlabeled DA. Twenty to 30% of the radioactivity accumulated could be released upon depolarization within a period of 5 min. At 6 DIV, influx of [3H]DA into diencephalic DA neurons was almost insensitive to benztropine, reserpine and unlabeled DA. Even after 13 DIV, diencephalic DA uptake was characterized by a markedly lower initial velocity, a longer time needed to reach saturation, a lower uptake capacity, and a lower sensitivity to benztropine than mesencephalic DA uptake. The releasable pool was very small and did not increase between DIV 6 and 13. The results demonstrate that mesencephalic DA neurons in vitro differentiate considerably faster than diencephalic DA neurons and gain functional competence very early in brain development. Comparison with data on adult nigrostriatal and hypothalamic DA systems suggests that the in vitro differences reflect a fundamental regional diversity of DA neurons. PMID- 2565149 TI - Immunohistochemical distribution of somatostatin in the infant hypothalamus. AB - Somatostatin (SS)-containing neurons were mapped in the normal infant hypothalamus with immunohistochemistry, using the peroxidase anti-peroxidase technique. Neurons displaying SS immunoreactivity show a widespread distribution throughout the hypothalamic region. Principal SS-immunoreactive like (SS-IL) perikarya are located in the paraventricular, infundibular and posterior nuclei and in the preoptic region. High SS innervation is also found in the ventromedial and in the lateral mammillary nuclei, and in the median eminence. In general this distribution of SS-IL agrees well with that reported for rat. Compared to the immunohistochemical distribution of SS in human adult hypothalamus, this mapping in the infant hypothalamus is grossly similar. However some differences may be underlined: the presence of a moderately dense group of SS-IL perikarya in the tuberal and posterior nuclei, and a dense innervation of the ventromedial nucleus and in the median eminence. This first detailed distribution of SS immunoreactivity in infant hypothalamus can provide basic knowledge for further studies of infant neuropathology. PMID- 2565150 TI - Glucose deprivation neuronal injury in cortical culture. AB - Murine cortical cell cultures deprived of glucose for 6-8 h developed extensive neuronal degeneration, apparent both morphologically and by efflux of lactate dehydrogenase to the bathing medium. This neuronal damage could be substantially reduced by addition of D-2-amino-5-phosphonovalerate (D-APV), in a concentration dependent (IC50 about 2 microM) and stereospecific (D-APV more potent than L-APV) fashion. A similar neuron-protective effect could also be obtained with several other NMDA antagonists, 2-amino-7-phosphonoheptanoate, phencyclidine, MK-801, ketamine, and (+)-SKF 10,047, as well as with the broad spectrum glutamine antagonist kynurenate. In contrast, little protection could be obtained with gamma-D-glutamylaminomethyl sulfonate and L-glutamate diethyl ester, compounds which have been reported to act primarily at non-NMDA receptors. These observations support the hypothesis that glucose deprivation-induced cortical neuronal injury is largely mediated by NMDA receptors, and suggest that cell culture methodology can be useful in the quantitative characterization of that injury. PMID- 2565151 TI - Electrophysiological and pharmacological properties of putative A13 incertohypothalamic dopamine neurons in the rat. AB - A13 incertohypothalamic dopamine (DA) neurons were labelled with antibodies raised to tyrosine hydroxylase in the male rat. Electrophysiologically, these neurons could be distinguished from their neighboring non-DA cells by their wide action potentials (greater than 2 ms), slow firing rates (0-3.8 impulses/s) and by the ability of iontophoresed DA and systemically administered apomorphine to inhibit impulse flow. Low doses of the antipsychotic drug haloperidol attenuated DA's response and reversed the apomorphine-inhibition of impulse flow. PMID- 2565152 TI - Excitatory effect of histamine on the arousal system and its inhibition by H1 blockers. AB - To clarify whether the sedative effect of H1 blockers is exerted in relation to H1 receptors in the brain, EEG activity recorded from the cortex and thalamus of rats was studied by power spectral analysis. EEG processing was performed by the FFT method and displayed as compressed spectral arrays. When a train of low frequency electrical stimulation was applied to the midbrain reticular formation of conscious rats, there was an increase in spectral power recorded at the cortex and thalamus, especially in the low frequency bands (0-6 Hz). The intraventricular administration of histamine suppressed the increase in power; this inhibition was antagonized by simultaneous administration of pyrilamine or diphenhydramine, though not in in combination with cimetidine or ranitidine. As in the case of histamine, the administration of 2-methylhistamine decreased power in the slow wave region, while administration of 4-methylhistamine did not. It was assumed that the arousal effect of histamine is exerted via H1 and not related to H2 receptors. Adverse effects of H1 blockers, such as drowsiness, may be caused by their inhibition of histamine's arousal effect. PMID- 2565153 TI - Beta-adrenergic and opioid receptors on pituicytes cultured from adult rat neurohypophysis: regulation of cell morphology. AB - Explants of adult rat neurohypophysis were maintained in culture for 14 days. The majority of cells present in the outgrowth of such cultures were identified as pituicytes on the basis of immunostaining for glial fibrillary acidic protein. Pituicytes were also stained by antisera to the membrane glycoprotein antigen Thy 1 and the extracellular matrix glycoprotein fibronectin. The cultures contained naloxone sensitive binding sites for the opioid receptor ligand [3H] dynorphin A 1-8 and peripheral-type benzodiazepine binding sites. Dynorphin binding was visualised over pituicytes following autoradiography. The morphology of cultured pituicytes was regulated by beta-adrenergic receptors present on the cells which, when activated, stimulated rapid transformation from a flattened irregular morphology to a stellate, process-bearing morphology. Dynorphin was without effect on the morphology of cultured pituicytes. These findings are discussed in the context of the known morphological plasticity of pituicytes in vivo. PMID- 2565155 TI - Penetrating wounds, abscesses, gravel, and bruising of the equine foot. AB - Foot-associated lamenesses are common diagnostic challenges for the equine practitioner. This article reviews the commonly occurring penetrating and concussive solar injuries of the equine foot. Diagnostic and therapeutic approaches for the management of these conditions are suggested. With proper treatment, many of these serious injuries will show excellent results. PMID- 2565154 TI - An electron microscopic study of somatostatin immunoreactive structures in lamina II of the rat spinal cord. AB - We are reporting the results of a light and electron microscopic study of somatostatin (SOM) immunoreactive (I) structures in lamina II of the lumbar spinal cord of the rat. At the light microscopic level, the observed distribution and morphology of SOM-I cell bodies and fibers confirmed published studies. At the electron microscopic level, SOM immunostaining in perikarya was localized to the golgi region. Immunostaining in cell bodies could be enhanced by colchicine treatment and after such treatment, it was noticeably increased in the cytoplasm. Synaptic contacts on SOM-I cell bodies were rare and SOM-I axons contacted unlabeled somata in lamina II. Some SOM-I dendrites participate in glomerular arrangements and they exhibited postsynaptic densities adjacent to the central profile of the glomerulus. Nonglomerular SOM-I dendrites and spines were postsynaptic to vesicles containing axons. Vesicle containing SOM-I axons presynaptic to larger dendrites were also observed in the outer portion of lamina II. Somatostatin has been implicated in nociception and some of the SOM-I structures reported here may be the anatomical substrates for SOM-induced analgesia. PMID- 2565156 TI - Management of lacerations and avulsion injuries of the foot and pastern region and hoof wall cracks. AB - The causes, clinical signs and various approaches to treatment of injuries involving the foot and pastern regions are reviewed, and the prognosis for each type of injury is discussed. PMID- 2565157 TI - Miscellaneous conditions of the equine foot. AB - The etiology, clinical presentation, radiographic findings, diagnostic criteria, differential diagnoses, treatment, and prognosis are reviewed for several clinically important conditions of the equine foot. These include pedal osteitis, sheared heels, distal sesamoid bone (navicular) fractures, subchondral bone cyst of the distal phalanx, distal interphalangeal joint subluxation, congenital phalangeal hypoplasia, bipartite and tripartite distal sesamoid bones, keratoma, ossification of the cartilages of the distal phalanx (sidebones), necrosis of the cartilages of the distal phalanx (quittor), thrush, canker, vesicular stomatitis, and chronic selenium toxicosis. PMID- 2565158 TI - Bevantolol attenuates thiazide stimulated renin secretion and catecholamine release in diuretic resistant hypertensives. AB - An attempt was made to establish whether cardioselective beta-blockade could counteract the stimulation by hydrochlorothiazide of the renin-angiotensin and sympathetic nervous systems and to what extent such actions contributed to the antihypertensive effect of bevantolol. The hemodynamic and neurohumoral responses of 21 thiazide resistant hypertensives who had received sequential chronic therapy with hydrochlorothiazide, hydrochlorothiazide combined with bevantolol and bevantolol monotherapy were compared. In these patients, bevantolol had a negative chronotropic effect and appeared, when administered alone, to induce an overall lowering of sympathetic nervous system activity without inhibiting the reflex responses of peripheral vascular resistance to postural change or lowered heart rate. When bevantolol and hydrochlorothiazide were administered together, sympathetic activity appeared to be maintained, possibly as a reflex response to volume depletion but vascular resistance did not appear to be responsive to baroreceptor stimulation. Diminished vascular reactivity induced by the hydrochlorothiazide is suspected to be a contributory factor. Inhibition of thiazide stimulated renin release by bevantolol may contribute to the antihypertensive effect of the combined therapy. PMID- 2565159 TI - DNA sampling and informed consent. AB - Standard consent forms for blood and tissue sampling are inadequate for DNA sampling. However, creating new and separate forms for each type of activity associated with DNA analysis (banking, linkage analysis and genetic diagnosis) tends to dissociate the participant from what is essentially a medical continuum. Furthermore, DNA sampling involves the sharing of samples and data among centres. To ensure patient control throughout this multifaceted process, we have developed an integrated approach to obtaining consent for DNA sampling at each level of participation. Movement from one level to another is reflected in the choices offered to participants. This inclusive approach is based on the underlying principle of informed consent, namely the respect for individuality, confidentiality and freedom of choice. This approach should help practitioners of medical genetics recognize the medical context of DNA sampling. PMID- 2565160 TI - Immunotherapy of murine hepatic metastases with lymphokine-activated killer cells expanded in serum-free media and recombinant interleukin 2. AB - It has been shown that the systemic administration of lymphokine-activated killer (LAK) cells with recombinant interleukin 2 (RIL-2) is effective in reducing the number of established pulmonary and hepatic metastases in murine models. Similarly, this modality of therapy has been proven effective against certain selected human tumors as well. In view of the rising concern with transmission of virally related communicable diseases such as hepatitis and AIDS, we have undertaken the evaluation of a serum-free medium (AIM V) for the generation and expansion of murine LAK cells for use in in vivo tumor immunotherapy against murine hepatic metastases. Day 3 LAK cells generated in AIM V medium demonstrated a greater percentage of viable cells than cells generated in serum containing complete medium (CM) (mean percentage of yield, 59 versus 25%, AIM V medium versus CM, respectively, P less than 0.001, N = 6 consecutive experiments). When day 3 LAK cells were transferred to new medium (CM to CM and AIM V to AIM V), a highly reproducible expansion of these cells was demonstrated which was significantly better for cells expanded in AIM V medium versus cells expanded in CM (mean fold expansion on day 21 of culture; 201 versus 54, AIM V medium versus CM, respectively, P less than 0.005, N = 4 consecutive experiments). When day 3 LAK cells, day 5 expanded LAK cells, and day 13 expanded LAK cells grown in CM or in AIM V medium were given in vivo with RIL-2 to mice harboring hepatic metastases, cells grown in AIM V medium demonstrated an increased antitumor activity compared to cells grown in CM. As an example in experiment 1, the mean number of metastases with day 5 expanded LAK cells grown in CM and given with RIL 2 was 47 while the mean number of metastases with day 5 expanded LAK cells grown in AIM V medium and given with RIL-2 was 5 (P less than 0.002). These experiments demonstrate that AIM V medium can be utilized to generate greater numbers of murine LAK cells with enhanced in vivo antitumor activity compared to cells generated in CM. These findings could be applied to the expansion of cytotoxic cells for human antitumor therapy. PMID- 2565161 TI - Positive inotropic/vasodilator agents. AB - Beta-adrenergic sympathomimetic agents such as dobutamine and dopamine, and phosphodiesterase inhibitors such as amrinone, milrinone, and enoxamone, exert a direct positive inotropic effect upon the myocardium by causing an increase in cyclic AMP levels. The phosphodiesterase inhibitors also exert a substantial direct vasodilator effect. Both the sympathomimetic agents and the phosphodiesterase inhibitors can be of value in the acute, short-term management of myocardial failure. At present, the use of these agents for long-term therapy of congestive heart failure is unproven, and remains investigational. PMID- 2565162 TI - Immunosuppression and beta-blockade in heart failure. AB - Two potential treatments, immunosuppression for myocarditis and beta-adrenergic blockade for heart failure, have been advocated. However, definitive conclusions await the result of ongoing large, multicenter, randomized clinical trials. There is circumstantial evidence that myocarditis may be a common precursor of dilated cardiomyopathy. Animal studies of myocarditis suggest that critically timed immunosuppression may be an effective therapy. Although promising, clinical studies to date are small and mostly uncontrolled. Clinical studies have demonstrated that beta-blockers are generally well tolerated in heart failure, and long-term treatment may result in improved symptomatic and functional status. These effects may be due to antagonism of direct and indirect effects of chronic reflex sympathetic stimulation on the cardiovascular system, including down regulation of the myocardial beta-adrenergic receptor. However, prolongation of survival has not been documented by any studies. Initiation of beta-blockade requires careful patient selection and titration with small initial doses of metoprolol. PMID- 2565163 TI - Neurohormonal influences in the pathogenesis of congestive heart failure. AB - Despite the interest in neurotransmitter and hormonal influences in heart failure in the last decade, the issues surrounding their contribution to the pathophysiology of congestive heart failure have become more complex. This must be considered in view of the pathway of hormonal stimulation and inhibition (Fig. 3) and the multiple steps where abnormal responses can interrupt these pathways. It is reasonable to state that the extent of neurotransmitter and hormonal activity in stage IV heart patients, treated with digoxin and diuretics, has been characterized. However, these observations cannot be extended to other stages of CHF, and assumptions cannot be made regarding either the untreated patient, or the nature of disease progression. Furthermore, there is insufficient information regarding the direct effect of these hormonal systems, with the exception of norepinephrine, on the myocardium. These issues must be the focus of future studies. PMID- 2565164 TI - Coexistence of multiple peptides in small intensely fluorescent (SIF) cells of inferior mesenteric ganglion of the guinea pig. AB - Coexistence of peptides in the small intensely fluorescent cells was demonstrated by immunocytochemistry for met-enkephalin-Arg-Gly-Leu, vasoactive intestinal polypeptide, somatostatin, neuropeptide Y and dynorphin. In the extreme example, a single cell was immunoreactive to all 5 peptides examined. Four peptides coexisted in 8% and three peptides in 13% of SIF cells. In 10% of SIF cells no peptide immunoreactivity could be detected. The most prevalent peptide was met enkephalin (in 46% of cells), then vasoactive intestinal polypeptide (45%), somatostatin (39%), neuropeptide Y (31%) and dynorphin (24%). Met-enkephalin and vasoactive intestinal polypeptide coexisted most commonly (25%). PMID- 2565165 TI - Vasoactive intestinal polypeptide-immunoreactive nerves in the pulmonary vasculature of the aquatic file snake Acrochordus granulatus. AB - The indirect immunofluorescence technique was used to determine the distribution of vasoactive intestinal polypeptide-immunoreactive and somatostatin immunoreactive axons in the pulmonary vasculature of the aquatic file snake Acrochordus granulatus. A dense distribution of vasoactive intestinal polypeptide immunoreactive axons was found on the common pulmonary artery, the anterior and posterior pulmonary arteries, and the smaller arteries branching to the lung. The density of these axons appeared greater in arterial preparations taken from more distal regions of the lung. The densest distribution of vasoactive intestinal polypeptide-immunoreactive axons was observed on the larger pulmonary veins in all regions of the lung. These axons were observed on the larger veins within the lung parenchyma but not on the smaller veins. Axons and cell bodies were observed in the vagal nerve trunks which run parallel to the pulmonary arteries and veins. In contrast, no somatostatin-immunoreactive axons were observed in any region of the pulmonary vasculature. It is proposed that the perivascular plexus of vasoactive intestinal polypeptide-immunoreactive axons may represent part or all of the vagal postganglionic innervation of the pulmonary vasculature. PMID- 2565166 TI - [Dipeptidylpeptidase IV in lymphocytes and lymphatic organs in humans]. PMID- 2565168 TI - Fentanyl, sufentanil, alfentanil and myocardial function. PMID- 2565169 TI - Fentanyl, sufentanil, alfentanil and myocardial function. PMID- 2565170 TI - Asystole after alfentanil-succinylcholine. PMID- 2565167 TI - Respiratory depression and spinal opioids. AB - Administration of epidural and intrathecal opioids may provide excellent postoperative analgesia, but a minority of patients will suffer dangerous respiratory depression. This review discusses the detection and measurement of respiratory depression and summarizes the relevant literature as it pertains to epidural and intrathecal opioid administration. The respiratory depressant effects and pharmacokinetics of spinal opioids are reviewed. The clinical implications and areas of future investigation are discussed. PMID- 2565171 TI - Further evidence that mitogen-induced cell proliferation does not support the formation of enzyme-altered islands in rat liver by carcinogens. AB - Our earlier studies have revealed that direct hyperplasia induced by liver mitogens such as lead nitrate, ethylene dibromide, nafenopin and cyproterone acetate, unlike compensatory cell proliferation induced by partial hepatectomy and CCl4, does not support the formation of enzyme-altered islands induced by chemical carcinogens in the liver. In the previous studies carcinogens were given at the peak of DNA synthesis induced by the liver mitogens. If the mitogens have altered the sensitive phase of the hepatocyte to the carcinogenic attack, administering the carcinogen at one time point following the mitogenic stimulus might have missed the sensitive phase. In order to overcome this possibility in the present study male Wistar rats weighing 200-250 g were given N-methyl-N nitrosourea (MNU; 60 mg/kg, i.p.) at three points representing G1, S and G2/M phases of the cell cycle following different types of liver cell proliferative stimuli. In another experiment MNU (60 mg/kg, i.p.) and diethylnitrosamine (15 mg/kg, i.p.) were given prior to the administration of proliferative stimuli. The initiated hepatocytes were also assayed following promotion by two different promoting regimens, namely phenobarbital and the resistant-hepatocyte model. Further, the initiated hepatocytes were monitored not only by using the appearance of islands of enzyme-altered hepatocytes but also using the incidence of hepatocellular carcinoma. The results of this study clearly revealed that irrespective of the protocol used, only the compensatory liver cell proliferation but not the mitogen-induced direct hyperplasia supported the formation and the growth of enzyme-altered islands in the liver induced by chemical carcinogens. PMID- 2565172 TI - Gamma-glutamyl transpeptidase activity in superficial exfoliated cells during hamster buccal pouch carcinogenesis. AB - The study of gamma-glutamyl transpeptidase (GGT) activity in transformed tissues has conventionally involved the killing of an animal or excision of the lesion, preventing longitudinal study. Oral exfoliative cytology allows longitudinal examination; however, it has been shown to be an unreliable criterion of malignancy. GGT activity has been demonstrated histochemically to involve the full thickness of the epithelium, including the keratin layer during carcinogenesis of hamster buccal pouch epithelium. This study correlates the GGT stained foci in tissue sections and the proportion of GGT-positive cells in a superficial smear during a 13-week regime of tri-weekly topical application of 7,12-dimethylbenz[a]-anthracene (DMBA) in mineral oil. GGT-positive cells were detected in smears 3 weeks after application of the carcinogen, coincident with GGT-positive foci in tissue sections involving the keratin layer. The proportion of GGT-positive cells in each experimental period increased during the first 7 weeks of the experiment and plateaued thereafter. The number of GGT-positive foci in tissue sections in each experimental period also increased during the experiment. GGT activity was not detected in either smears or tissue sections of untreated or mineral oil treated mucosa. The correlation between the proportion of GGT-positive cells in smears and GGT-positive foci in tissue sections suggest the possibility of studying the GGT activity in an experimentally induced lesion without its elimination. Furthermore, oral exfoliative cytology using GGT staining may be useful in detecting precancerous lesions clinically. PMID- 2565174 TI - Free tissue transfer for reconstruction of the weight-bearing portion of the foot. AB - Twenty-eight patients treated with free tissue transfer for soft-tissue coverage of the weight-bearing portion of the foot were investigated over a minimum follow up period of 12 months. Seven were treated with a sensory innervated flap transfer, and 21 had a noninnervated skin or muscle flap transfer. Secondary procedures to debulk or sculpture the flap were necessary in seven patients. Complications occurred in six patients and were either related to lack of protective sensibility (in three) or excessive flap mobility (in three). Impaired flap sensibility was more common in noninnervated flaps, but excessive flap mobility was more typical of cutaneous flaps. The merits of an innervated cutaneous flap, which may be excessively bulky, must be weighed against those of a well-contoured muscle flap without sensory innervation. PMID- 2565173 TI - Comparison of the properties of [3H]-D-2-amino-5-phosphonopentanoic acid and [3H] DL-2-amino-7-phosphonoheptanoic acid binding to homogenates of rat cerebral cortex. AB - 1. The pharmacology and ionic regulation of [3H]-2-D-2-amino-5-phosphonopentanoic acid ([3H]-D-AP5) and [3H]-DL-2-amino-7-phosphonoheptanoic acid ([3H]-DL-AP7) binding to homogenates of rat cerebral cortex were examined using radioligand binding methodology. 2. Both [3H]-D-AP5 and [3H]-DL-AP7 labelled a single population of binding sites with dissociation constants of 0.39 and 1.8 mumol/l, respectively. The density of binding sites found with [3H]-DL-AP7 was 13 times greater than that found with [3H]-D-AP5. 3. The ionic requirements of the [3H]-D AP5 binding site in the presence of chloride were such that calcium acetate enhanced binding, while magnesium and sodium acetate both decreased binding. In the absence of chloride both calcium and chloride ions stimulated binding. 4. In a chloride-free buffer calcium acetate stimulated binding of [3H]-DL-AP7 in a biphasic manner. Chloride ions (ammonium salt) enhanced binding slightly at low concentrations (0.1-1.0 mmol/l) above which binding was reduced to non-specific levels. The ionic dependence of [3H]-DL-AP7 binding had some similarities to the previously defined GLU-C site. 5. The pharmacological profile of the site labelled by [3H]-D-AP5 was consistent with that of a recognition site for N methyl-D-aspartate (NMDA) as defined in electrophysiological experiments. [3H]-DL AP7 did not label an NMDA site as several non-NMDA ligands displaced binding with high affinity and the binding was not stereospecific as found for [3H]-D-AP5. Moreover, the pharmacological profile of the [3H]-DL-AP7 site did not correspond to any excitatory amino acid receptor as presently defined. PMID- 2565175 TI - The radionuclide assessment of a system for slow intrathecal infusion of drugs. AB - The authors describe a new radionuclide method for assessing the functional integrity of slow intrathecal infusion devices. Approximately 11 mCi (400 MBq) of Tc-99m DTPA is injected into the pump chamber. Early and delayed imaging of the pump, tubing, and spinal cerebrospinal fluid space is performed on a digital large-field-of-view gamma camera. The digitally displayed images are then reviewed by adjustment of the grey scale window. Four patients have had eight studies and in each case the result has been confirmed by surgical exploration or by clinical response to change in therapy. PMID- 2565177 TI - Hyperactivity and inattentiveness. School assessment of stimulant treatment. AB - The school health folders of all students receiving stimulant medication for hyperactivity/inattentiveness from five public elementary and two middle schools in Baltimore County were evaluated to 1) record pre-treatment teacher ratings of those on stimulant medication; 2) assess, using the rating scales, the initial and subsequent effectiveness of the medication; and 3) to identify treatment subgroups. Major findings were 1) three-fourths of the 176 students on stimulant medication in 1987 had pre-treatment ratings consistent with moderate to severe hyperactivity/inattentiveness; 2) over 90 percent of the students evidenced at least 50 percent improvement initially following stimulant treatment by teacher ratings; 3) 76 percent of the medicated students continued to show this level of improvement at the end of the 1987 school year; 4) medication improvement and compliance declined in middle school; 5) 15 percent of the students on stimulant medication in 1987 were inattentive but not hyperactive on baseline teacher ratings, but their degree of improvement with stimulants was equivalent to that of hyperactives; 6) in the county-run hyperkinetic clinic, the subpopulation of inattentive/non-hyperactive students on stimulants rose from 7 percent to 18 percent of the total between 1976 and 1987. PMID- 2565178 TI - Effect of alpha-2 adrenergic receptor stimulation on short-circuit current across isolated skin of the toad Bufo arenarum. AB - 1. The effect of the alpha-2 adrenergic agonist clonidine on short-circuit current (SCC) across isolated skins of Bufo arenarum toads was investigated. 2. Clonidine inhibited basal SCC in a dose-dependent manner. 3. Blockade of the effect of clonidine on basal SCC by the selective alpha-2 antagonist yohimbine supports the hypothesis that the inhibitory effect is mediated by the stimulation of alpha-2 adrenergic receptors. 4. The fact that the inhibitory effect of clonidine is higher in skins with spontaneous positive SCC than in the negative ones, and that the alpha-2 agonist was unable to alter amiloride-induced negative SCC suggests that the inhibitory effect of clonidine may probably be mediated by inhibition of sodium transport. PMID- 2565179 TI - Induction of glutathione S-transferases in genetically inbred male mice by dietary ethoxyquin hydrochloride. AB - 1. Constitutive and ethoxyquin hydrochloride (EQ-HCl)-induced hepatic glutathione (GSH) S-transferase, GSH reductase, and GSH peroxidase activities were determined in 5 strains of 8-10 week old inbred male mice. 2. The constitutive GSH S transferase (GST) activity varied from 2.9 (SJL/JCR) to 8.9 (C57BL/6NCR) mumol product formed/min/mg protein and the corresponding values for the EQ-HCl-treated mice were in the range of 15.3-25.3 mumol product formed/min/mg protein. 3. EQ HCl induced GST activity in all the strains examined and this contrasted to the induction activity of Aroclor 1254 which was strain-dependent. GST activity was induced 2.9-fold in Aroclor 1254-responsive (C57BL/6) and 2.8-fold in non responsive (DBA/2) mice, respectively. PMID- 2565180 TI - The venom of Ampulex compressa--effects on behaviour and synaptic transmission of cockroaches. AB - 1. The solitary wasp Ampulex compressa stings a cockroach, Periplaneta americana, twice. 2. The first sting into the ventral thorax results in a transient paralysis. During this paralysis the wasp stings the suboesophageal ganglion, which gradually results in a permanent deactivation. 3. The venom gland is a paired and highly branched organ, with a common ductus venatus. The large lumen is lined with a folded cuticula. No venom reservoir is present. 4. Extract of the venom gland induces a slow contraction of the guinea pig ileum. 5. The agonist present in the venom cannot be identified with a known agonist. 6. Venom gland extract blocks synaptic transmission from the cercal nerve to giant neurons in the sixth abdominal ganglion of the cockroach. 7. The block develops gradually, like the gradual appearance of the effects of the sting into the suboesophageal ganglion on the behaviour of the cockroach. PMID- 2565181 TI - Excitotoxicity, reflex responses, and evoked changes in extracellular potassium in the frog spinal cord. AB - 1. The effects of the excitatory amino acid agonists kainate (KA), quisqualate (QUIS), and N-methyl-D-aspartate (NMDA) were studied in vitro on the hemisected frog spinal cord. 2. Prolonged (1.0 hr) application of excitatory amino acid agonists (KA, 50 or 300 microM; QUIS, 30 microM; NMDA, 300 microM) significantly reduced the ventral root potentials (VRPs) and [K+]0 evoked by a dorsal root tetanus (10 sec, 25 Hz), by brief (10 sec) applications of the same agonists (KA, 30 microM; QUIS, 30 microM; NMDA, 300 microM), and by GABA (10 sec, 1.0 mM). 3. The effect was essentially irreversible and persisted despite 2-4 hr of washing. 4. Excitatory amino acid antagonists (APV, 30 microM and kynurenate, 2 mM) blocked the neurotoxic effects of the excitatory agonists NMDA and KA respectively, an observation which indicates the observed effects of the agonists require the activation of specific excitatory receptors. 5. TTX did not alter the neurotoxic effects of KA suggesting that interneuronal firing does not contribute to the observed changes. 6. Addition of high K+ did not duplicate the effect of prolonged excitatory amino acid agonist exposure, an indication that elevation of K+ does not cause the decreased responses. 7. Light microscopy did not provide any evidence of gross tissue damage. 8. The parallel reduction of postsynaptic responses and delta [K+]0 support the idea that elevation of extracellular [K+] by afferent stimuli results from interneuronal activity. PMID- 2565182 TI - Further consideration of phenobarbital effects on cytochrome P-450 activity in the killifish, Fundulus heteroclitus. AB - 1. Ethoxyresorufin O-deethylase (EROD) activity, aldrin epoxidase (AE) activity, cytochrome P-450 content, and levels of cytochrome P-450E (the major BNF inducible P-450 form and primary EROD catalyst in scup) or its homologues were measured in hepatic microsomes isolated from Fundulus heteroclitus, scup (Stenotomus chrysops) and brook trout (Salvelinus fontinalis) treated with beta naphthoflavone (BNF) or phenobarbital (PB). 2. In all three teleost species, BNF treatment caused expected increases in P-450 content, EROD activity and P-450E level; but either no change or a slight decrease in AE turnover rate (nmol/min/nmol P-450). 3. Polyclonal antibodies to P-450E did not inhibit AE activity in microsomes from BNF-treated scup, confirming that this major BNF inducible P-450 form does not catalyze AE activity in fish. 4. In contrast, PB treatment did not affect hepatic AE activity, P-450 content or levels of "P-450E" in F. heteroclitus, but did variably affect EROD activity which was suppressed in one experiment and elevated in another. 5. The results indicate that (i) contrary to previous reports, neither PB nor MC-type inducers increase AE activity in F. heteroclitus, (ii) MC-type inducers do not affect AE activity in the other teleost species examined, and (iii) AE activity is not a reliable indicator of P 450 induction by environmental chemicals. 6. We emphasize the need to establish the mechanism of PB action, and the nature of any fish P-450 forms analogous to PB-inducible forms in mammals in order to conclusively evaluate PB-responses in fish. PMID- 2565176 TI - Clinical pharmacokinetics of non-opiate abused drugs. AB - The present review discusses the available data on the kinetic properties of non opiate abused drugs including psychomotor stimulants, hallucinogens and CNS depressants. Some of the drugs of abuse reviewed here are illicit drugs (e.g. cannabis, cocaine), while others are effective pharmacological agents but have the potential to be abused (e.g. benzodiazepines). Although some of the drugs mentioned in this review have been in use for centuries (e.g. caffeine, nicotine, cocaine, cannabis), knowledge of their kinetics and metabolism is very recent and in some cases still incomplete. This is partially due to the difficulties inherent in studying drugs of abuse in humans, and to the complex metabolism of some of these drugs (e.g. cannabis, caffeine) which has made it difficult to develop sensitive assays to determine biological pathways. Although drugs of abuse may have entirely different intrinsic pharmacological effects, the kinetic properties of such drugs are factors contributing to abuse and dependence. The pharmacokinetic properties that presumably contribute to self-administration and drug abuse include rapid delivery of the drug into the central nervous system and high free drug clearance. Kinetic characteristics also play an important role in the development of physical dependence and on the appearance of a withdrawal syndrome: the longer the half-life, the greater the likelihood of the development of physical dependence; the shorter the half-life, the earlier and more severe the withdrawal. The balance between these 2 factors, which has not yet been carefully studied, will also influence abuse patterns. The clinical significance of kinetic characteristics with respect to abuse is discussed where possible. PMID- 2565183 TI - Comparative studies on the distribution of rhodanese and beta-mercaptopyruvate sulfurtransferase in different organs of sheep (Ovis aries) and cattle (Bos taurus). AB - 1. The activities of rhodanese and beta-mercaptopyruvate sulfurtransferase (MST) in different organs of sheep and cattle were measured. 2. Liver, kidney, omasum, and rumen were the richest sources of both enzymes. The activities of both enzymes in other organs of the sheep and the cattle decreased in the order of lung, brain, heart, abomasum, lymph node, urinary bladder, spleen, and the skeletal muscle. 3. The activities of both enzymes in most organs of the sheep were higher than the cattle. 4. Both enzymes showed higher activities in the epithelial layers than the muscular layers of rumen, omasum and reticulum. 5. In most of the tissues of both species the level of rhodanese activity was greater than MST. PMID- 2565184 TI - Elevation of sex steroids and inhibition of UDP-glucuronyltransferase are out of phase during gonadal maturation in the common carp. AB - 1. Plasma sex steroid concentrations, onset of gonadal maturation, and hepatic microsomal UDP-glucuronyltransferase (UDPGT) activities were followed under natural temperature and photoperiod in outdoor tanks, and under controlled laboratory temperature and photoperiod regimens in common carp (Cyprinus carpio). 2. Decreased activity of UDPGT was out of phase with elevations in plasma testosterone and 17 beta-estradiol during gonadal maturation. 3. Injection of pituitary extract induced final gonadal maturation and transient elevations (within 24 hr) of both plasma sex steroid concentrations and UDPGT activities. 4. There were no simple relationships between plasma sex steroid concentrations and activity of hepatic microsomal UDPGT in common carp. PMID- 2565185 TI - Two subtypes of acetylcholinesterase isoenzymes distinguishable by Angusticeps type toxin F7. AB - 1. Toxin F7, a toxin isolated from Dendroaspis angusticeps (green mamba) venom, exhibited a potent inhibition on the acetylcholinesterase of Bungarus snake venoms and homogenized brains and muscles of Bungarus and Trimeresurus snakes as well as that of mammalian tissues. 2. The acetylcholinesterase in the venoms and tissues of Naja (cobra) species as well as that in avian tissues, however, was found to be about 1000 times less susceptible towards inhibition by toxin F7. 3. It is concluded that there exist at least two subtypes of acetylcholinesterase isoenzymes distinguishable by an angusticeps-type toxin F7. PMID- 2565186 TI - Effects of hypothalamic releasing hormones and biogenic amines on identified neurones in the circumoesophageal ganglia of the water snail (Planorbis corneus). AB - 1. The effect of locally applied releasing hormones, thyrotropin-releasing hormone (TRH) and luteinizing-hormone-releasing hormone (LHRH) and the putative neurotransmitters, acetylcholine (ACh) and dopamine (DA), on the neuronal excitability of identified invertebrate giant dopaminergic neurone (GDN) and serotoninergic neurone (5-HT) (Planorbis corneus) were investigated by intracellular recording in vitro. 2. The membrane potential of GDN was of the order of -60 to -70 mV. The microiontophoretically applied substances produced membrane depolarization as well as spike activation. Their order of efficacy was as follows: TRH greater than ACh greater than DA greater than LHRH. 3. The effects of the tested TRH, ACh, LHRH and DA on serotoninergic neurones were less pronounced. 4. During ACh depolarization the membrane resistance of GDN was found to be strongly reduced, whereas TRH produced only a small reduction in membrane resistance. 5. Dihydro-beta-erythroidin (DHE) added to the bath solution reversibly blocked ACh depolarization without influencing TRH depolarization. Concentrations of atropine sulfate required to block the ACh depolarization were higher by at least 100 order of magnitude. 6. These effects are discussed in relation to the immunoreactive TRH detected earlier in the central nervous system of invertebrates and vertebrates. The results are consistent with the postulate that TRH acts as a neuromodulator and/or neurotransmitter on invertebrate and vertebrate neurones. PMID- 2565187 TI - The effect of yohimbine on plasma levels of T3, T4 and cortisol in xylazine immobilized white-tailed deer. AB - 1. The effect of yohimbine (Y) on blood levels of thyroxine (T4), triiodothyronine (T3), and cortisol was investigated in 5 mature male white tailed deer immobilized with xylazine hydrochloride (X). 2. T4 levels were erratic in X-treated deer, but stabilized in the X- and Y-treated deer. 3. T3 remained unchanged in both groups. 4. Cortisol levels have increased in X-treated deer, but declined in X- and Y-treated deer. 5. Yohimbine is a potent and safe antidote of X not affecting T3 and T4. Caution should be used in using R or Y in cortisol studies. PMID- 2565188 TI - Further studies on the understanding of Octodon degus natural resistance to morphine: a comparative study with the Wistar rat. AB - 1. Octodon degus shows higher levels of tolerance to morphine when compared with the Wistar rat. 2. In the formalin algesiometric test, this caviomorph is more resistant to pain (P less than 0.01) and to the analgesic effect of morphine (P less than 0.001). 3. CD50 and LD50 were significantly higher in Octodon degus as compared with Wistar rat. 4. Morphine caused in rat severe hypotension, while doses eight times higher in O. degus had a transient effect. 5. 3H-naloxone binding in adrenal glands of O. degus is higher than in other tissue samples assayed from the same animal or rats. PMID- 2565189 TI - Intraocular pressure in cats is lowered by drops of hornet venom. AB - 1. Nine cats were given an intravenous injection of the Oriental hornet (Vespa orientalis, Vespinae; Hymenoptera) venom sac extract (VSE) and seven cats had the same VSE administered as eye drops. 2. When injected intravenously, the hornet VSE decreased the intraocular pressure in both eyes sharply during the first 20 min and with a slower rate later on until the end of the 3 hr experiment. The intraocular pressure dropped to zero in some cases. 3. VSE eye drops decreased the intraocular pressure only in the treated eye, while in the second eye (left as a control) the intraocular pressure remained the same throughout the experiment. 4. The decrease in the intraocular pressure was sharp during the first 20 min and slowed down afterwards until the end of the experiment. 5. The intraocular pressure did not reduce to zero. 6. This study shows that the active components of the hornet venom which caused a decrease in the intraocular pressure can cross the cornea and exert a hypotensive effect in the eye. PMID- 2565190 TI - Interaction of serotonin and leu-enkephalin on the habituating central neurons of Helix pomatia L. in situ and in vitro. AB - 1. Habituating neurons (a, b and c) of Helix pomatia reacted to the serotonin (10(-5)-10(-4)M) with depolarization evoking oscillatory waves and burst firing at the range of -35 to -55 MP values. 2. Isolated habituating cells were hyperpolarized by serotonin and failed to generate membrane oscillation or bursting pattern. 3. Only the isolated habituating neurons reacted to the application of leu-enkephalin (10(-5)-10(-4)M) by depolarization. 4. Neither membrane oscillation nor burst firing were evoked by leu-enkephalin. 5. On the cells a, b and c leu-enkephalin modulated the serotonin effect through cyclic 3',5'-AMP system both in situ and in vitro. 6. The membrane oscillation and burst firing of the habituating cells are connected to the regulation of various rhythmic processes including pneumostoma movements. PMID- 2565191 TI - Glucuronides in mussel Mytilus galloprovincialis as a possible biomonitor of environmental carcinogens. AB - 1. The in vitro incubation of mussel digestive gland with 1 mM aminofluorene resulted in the formation of glucuronides that (a) became mutagenic with carp liver S9, and (b) liberated S9-dependent mutagenic aglucones after beta glucuronidase treatment. 2. Natural populations of mussels from unpolluted and polluted sites, as well as mussels exposed to 3 ppm of aminofluorene or to used engine oil, did not accumulate detectable amounts of premutagens, mutagens, or mutagenic glucuronides/aglucones either in digestive gland tissue or in shell cavity water. 3. The mutagenicity testing of mussel's glucuronides/aglucones does not seem to be useful as a biomonitor of environmental carcinogens. PMID- 2565192 TI - Effect of mazindol on dystrophic mice and on growth in young rats. AB - 1. Mazindol, which has been proposed as a therapy for muscular dystrophy because of a suppression of growth hormone release was administered orally (0.1 mg/kg body wt/day) for approximately six weeks to healthy young rats and dystrophic mice. 2. Mazindol had no effect on the dystrophic mice. 3. Mazindol treated rats had reduced wt gain, but this effect was due to appetite suppression not growth hormone inhibition. 4. No effect of mazindol was seen on rat muscle, but there were significant increases in liver, heart and kidney wts compared to controls. PMID- 2565193 TI - Some characteristics of mitochondrial monoamine oxidase activity in eggs of carp (Cyprinus carpio) and rainbow trout (Salmo gairdneri). AB - 1. Monoamine oxidase (MAO) activity towards tryptamine, 5-hydroxytryptamine (5 HT) and phenylethylamine (PEA) has been measured in mitochondria isolated from carp and trout eggs. 2. In carp eggs all the tested substrates are metabolized and the highest affinity is found with tryptamine. In trout eggs a consistent level of MAO activity is obtained using tryptamine. 3. The inhibition dose response curves of clorgyline and deprenyl indicate that both in carp and trout eggs there is only one form of mitochondrial MAO, distinct from MAO A and B which have been described in vertebrate tissues. 4. Both in carp and trout egg mitochondria a semicarbazide-sensitive amine oxidase is not involved in the deamination of the used substrates. 5. MAO found in carp and trout eggs might be involved in metabolism of some neurotransmitter monoamines during early developmental stages. PMID- 2565194 TI - Responses of the three-toed sloth, Bradypus tridactylus, to some commonly used pharmacologic agents. II. Chloralose and reserpine. AB - 1. Chloralose, 50 mg/kg i.v., is a safe effective anesthetic for sloths and reduces incidence of cardiac arrhythmias. 2. However, chloralose blocks baroreflexes and may reduce the sensitivity of beta 1 cardiac receptors. 3. Reserpine, 0.70 mg/kg given i.v. in divided doses, blocks the hypertensive effect of 100 micrograms/kg of tyramine in sloths. 4. Reserpine in this dosage materially reduces arterial pressure and heart rate; these effects last at least 7 days. 5. Reserpine potentiates the hypertensive effects of epinephrine and norepinephrine materially. 6. In sloths reserpine increases cardiac irritability but does not block baroreflexes. 7. As is true with most other drugs sloths are more sensitive to chloralose and reserpine than most common laboratory animals. PMID- 2565195 TI - Effect of aging on presynaptic alpha 2-adrenoceptor mechanisms in guinea pig ileum. AB - 1. Presynaptic alpha 2-adrenoceptor mechanisms in electrically stimulated longitudinal muscles of ilea isolated from 3, 10, 20 and 47 week-old guinea pigs were studied by analysis of the concentration-response curves of noradrenaline, a full agonist, and clonidine, a partial agonist, and the Scatchard plot of specific binding of [3H]-p-aminoclonidine to synaptosomal fractions from the longitudinal muscle of guinea pig ileum. 2. The pD2 value of noradrenaline and the maximum contraction induced by clonidine increased with age from 3 to 20 weeks and there after decreased to 47 weeks, while the pA2 value of yohimbine against noradrenaline did not alter with age. 3. The capacity of the maximum binding sites of [3H]-p-aminoclonidine increased with increasing age (3-20 weeks), while the dissociation constant (Kd) of [3H]-p-aminoclonidine did not change during the same period. 4. The changes in the presynaptic alpha 2 adrenoceptor mechanisms with age are considered to be due to the change in the total concentration of presynaptic of alpha 2-adrenoceptors. PMID- 2565196 TI - Alpha 2-adrenergic and VIP receptors in rabbit ciliary processes interact. AB - The interaction between alpha 2-adrenergic and VIP receptors has been studied by examining inhibition of VIP-stimulated cyclic AMP production by adrenergic agonists in intact, excised rabbit ciliary processes. Epinephrine, norepinephrine, isoproterenol, dopamine, and the specific alpha 2-adrenergic agonists clonidine and p-aminoclonidine exhibit dose-dependent inhibition of VIP stimulated cyclic AMP production. I50s, clonidine (0.05 microM) = p aminoclonidine (0.05 microM) congruent to epinephrine (0.1 microM) less than norepinephrine (2.0 microM) less than isoproterenol (15 microM) = dopamine (15 microM), are consistent with the characteristic binding affinities of these adrenergic agonists for alpha 2-adrenergic receptors. Inhibition of VIP stimulated cyclic AMP production by clonidine, epinephrine, isoproterenol, and dopamine is blocked by yohimbine but not by prazosin. These data establish the alpha 2-adrenergic specificity of the inhibitory effects observed. We have previously shown that beta 2-adrenergic receptor-mediated stimulation of cyclic AMP production in rabbit ciliary processes is also inhibited by postjunctional alpha 2-adrenergic receptors. These studies support the idea that the catecholamines may regulate aqueous humor formation by inhibiting stimulation of cyclic AMP production via postjunctional alpha 2-adrenergic receptors in ciliary processes. PMID- 2565197 TI - Inhibition of calcium of beta adrenoceptor mediated cAMP responses in isolated bovine corneal epithelial cells. AB - The increases in adenosine 3',5' monophosphate (cAMP) content were measured in isolated bovine corneal epithelial cells in response to either adrenergic agonists or adenylate cyclase stimulation. The beta selective adrenergic agonist, isoproterenol, and the adrenergic agonists, norepinephrine as well as epinephrine elicited large increases in cAMP accumulation. At their maximum effective concentrations, the respective increases were 16-fold, 6.6-fold and 4.7-fold. These stimulatory effects were completely inhibited by the beta selective adrenergic antagonist, propranolol. Similarly forskolin increased cAMP content more than 3-fold. These increases and the previous identification of beta adrenoceptors in fresh intact bovine corneas as well as cells in culture indicate that this enzymatic dissociation procedure does not affect the cAMP responses to either adrenergic agonists or forskolin. The relationship was considered between increases in Ca2+ concentration and the effects of either isoproterenol or forskolin, on cAMP accumulation. There were no changes in any of the cAMP responses at bathing solution Ca2+ concentrations between 0.01 microM and 1 microM. However, in cells permeabilized to Ca2+ with 10 microM ionomycin, increases within this concentration range depressed the baseline levels of cAMP content. Furthermore, the stimulatory effects of both forskolin and isoproterenol on cAMP accumulation were significantly blunted in this concentration range. These blunting effects by Ca2+ were not the result of any measurable decrease in ATP content. This negative relationship between increases in Ca2+ concentration and increases in cAMP content indicates that changes in intracellular Ca2+ concentration could modulate the second messenger function of cAMP linked to these agents. PMID- 2565199 TI - Hypertension. AB - An estimated 58 million Americans are at increased risk of morbidity and premature death due to high blood pressure (BP) and require some type of therapy or systematic monitoring. This article focuses on recent advances in our understanding of the pathogenesis of hypertension, new approaches to the diagnosis and treatment of secondary hypertension, and current views of the most appropriate nonpharmacologic and pharmacologic therapy for essential hypertension. In view of the extremely high prevalence of the disorder, emphasis is placed on efficient and cost-effective strategies for diagnosing and managing the hypertensive patient. Recent evidence indicates that nonpharmacologic therapy, including dietary potassium and calcium supplements, reduction of salt intake, weight loss for the obese patient, regular exercise, a diet high in fiber and low in cholesterol and saturated fats, smoking cessation, and moderation of alcohol consumption produces significant sustained reductions in BP while reducing overall cardiovascular risk. Accordingly, nonpharmacologic antihypertensive therapy should be included in the treatment of all hypertensive patients. In persons with mild hypertension, nonpharmacologic approaches may adequately reduce BP, thereby avoiding the expense and potential side effects of drug therapy. In patients with more severe hypertension, nonpharmacologic therapy, used in conjunction with pharmacologic therapy, can reduce the dosage of antihypertensive medications necessary for BP control. Patients treated with nonpharmacologic therapy only should be followed closely, and if BP control is not satisfactory, drug therapy should be added. The large number of drugs available for use in hypertension treatment, coupled with our rapidly expanding knowledge of the pathophysiology of hypertension and of the adverse effects of these drugs in individual patient groups, make it possible to individualize antihypertensive treatment. When used as monotherapy, most agents effectively lower BP in the majority of patients with mild or moderate essential hypertension. Thus, a single agent from one of four classes: diuretics, angiotensin-converting enzyme inhibitors, calcium channel blockers, and beta adrenergic blockers, usually provides effective BP control with minimal side effects in most patients. Therapy should be initiated with the agent most likely to be effective in BP lowering and best tolerated. If the initial agent is ineffective at maximal recommended therapeutic doses or has undue side effects, an alternative agent from another class should be tried. When monotherapy is unsuccessful, a second agent, usually of a different mechanism of action, should be PMID- 2565198 TI - Studies of He-T DNA sequences in the pericentric regions of Drosophila chromosomes. AB - He-T DNA is a complex set of repeated DNA sequences with sharply defined locations in the polytene chromosomes of Drosophila melanogaster. He-T sequences are found only in the chromocenter and in the terminal (telomere) band on each chromosome arm. Both of these regions appear to be heterochromatic and He-T sequences are never detected in the euchromatic arms of the chromosomes (Young et al. 1983). In the study reported here, in situ hybridization to metaphase chromosomes was used to study the association of He-T DNA with heterochromatic regions that are under-replicated in polytene chromosomes. Although the metaphase Y chromosome appears to be uniformly heterochromatic, He-T DNA hybridization is concentrated in the pericentric region of both normal and deleted Y chromosomes. He-T DNA hybridization is also concentrated in the pericentric regions of the autosomes. Much lower levels of He-T sequences were found in pericentric regions of normal X chromosomes; however compound X chromosomes, constructed by exchanges involving Y chromosomes, had large amounts of He-T DNA, presumably residual Y sequences. The apparent co-localization of He-T sequences with satellite DNAs in pericentric heterochromatin of metaphase chromosomes contrasts with the segregation of satellite DNA to alpha heterochromatin while He-T sequences hybridize to beta heterochromatin in polytene nuclei. This comparison suggests that satellite sequences do not exist as a single block within each chromosome but have interspersed regions of other sequences, including He-T DNA. If this is so, we assume that the satellite DNA blocks must associate during polytenization, leaving the interspersed sequences looped out to form beta heterochromatin. DNA from D. melanogaster has many restriction fragments with homology to He-T sequences. Some of these fragments are found only on the Y. Two of the repeated He-T family restriction fragments are found entirely on the short arm of the Y, predominantly in the pericentric region. Under conditions of moderate stringency, a subset of He-T DNA sequences cross-hybridizes with DNA from D. simulans and D. miranda. In each species, a large fraction of the cross-hybridizing sequences is on the Y chromosome. PMID- 2565200 TI - Methoxyflurane enhances allyl alcohol hepatotoxicity in rats. Possible involvement of increased acrolein formation. AB - The effect of methoxyflurane anesthesia on allyl alcohol-induced hepatotoxicity and the metabolism of allyl alcohol was studied in male rats. Hepatotoxicity was assessed by the measurement of serum alanine aminotransferase activity and histopathological examination. Allyl alcohol-induced hepatotoxicity was enhanced when allyl alcohol (32 mg/kg) was administered 4 hr before or up to 8 days after a single 10-min exposure to methoxyflurane vapors. The possibility that methoxyflurane increases alcohol dehydrogenase-dependent oxidation of allyl alcohol to acrolein, the proposed toxic metabolite, was evaluated by measuring the rate of acrolein formation in the presence of allyl alcohol and liver cytosol. The effect of methoxyflurane on alcohol dehydrogenase activity in liver cytosol was also assessed by measuring the rate of NAD+ utilization in the presence of ethyl alcohol or allyl alcohol. Alcohol dehydrogenase activity and rate of acrolein formation were elevated in methoxyflurane-pretreated rats. The results suggest that a modest increase in alcohol dehydrogenase activity and rate of acrolein formation markedly enhances allyl alcohol-induced hepatotoxicity. PMID- 2565201 TI - The covalent binding to protein of valproic acid and its hepatotoxic metabolite, 2-n-propyl-4-pentenoic acid, in rats and in isolated rat hepatocytes. AB - The covalent binding of radioactivity to protein following administration of 14C labeled analogs of valproic acid (VPA) and a hepatotoxic metabolite thereof, 2-n propyl-4-pentenoic acid (delta 4-VPA), was investigated in male rats. Covalent binding occurred in a number of tissues, the level of binding being greatest to proteins in liver for each compound. Moreover, the binding of radioactivity from delta 4-VPA to hepatic macromolecules was higher than the corresponding value for VPA. When radiolabeled VPA and delta 4-VPA were incubated with rat hepatocytes, radiolabel again became bound to cellular proteins, the time-course of which suggested the existence of at least two underlying mechanisms. Thus, after initial rapid binding of both substrates, a secondary slow phase was evident, which favored binding of delta 4-VPA. Although phenobarbital pretreatment of rats had little effect on the covalent binding of either substrate to isolated hepatocytes, clofibrate pretreatment markedly enhanced the covalent binding of both VPA and delta 4-VPA to these cells. In contrast, the covalent binding of VPA and delta 4-VPA was suppressed strongly by 4-pentenoic acid, a potent inhibitor of beta-oxidation, but was not affected by metyrapone, an inhibitor of cytochrome P-450 activity. (-)-Borneol and 8-bromo-cAMP, two inhibitors of glucuronidation, acted to decrease the binding of both substrates, although this inhibition was evident only in the early stages of incubation. A similar effect was seen with valeric acid, the saturated analog of 4-pentenoic acid.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565202 TI - Role of pharmacokinetics and metabolism in the enhanced susceptibility of middle aged male Sprague-Dawley rats to acetaminophen nephrotoxicity. AB - Middle-aged male Sprague-Dawley (SD) rats (9-12 months) are more susceptible to acetaminophen (APAP)-induced nephrotoxicity than are young (2-3 months) adult males. The present studies were designed to evaluate the role of pharmacokinetics and renal and hepatic metabolism of APAP in age-dependent nephrotoxicity. Following 750 mg/kg APAP, ip, a nephrotoxic dosage in 12-month-old but not 3 month-old rats, renal cortical APAP concentrations were significantly greater in 12-month-old compared with 3-month-old SD rats at 3, 4, and 6 hr after treatment. Renal medullary APAP concentrations in 12 month-old rats were significantly greater than in 3-month-old rats at 2, 3, and 5 hr after treatment. Serum APAP concentrations were significantly elevated in 12-month-old compared with 3-month old rats from 2 through 5 hr after APAP (750 mg/kg ip). However, APAP tissue/serum concentration ratios were similar in 3- and 12-month-old rats, indicating that differences in tissue concentration were secondary to increased serum concentrations in older rats. Conjugated APAP metabolites in blood were similar in 3- and 12-month-olds during the initial 2-3 hr after 750 mg/kg APAP, ip, but began to accumulate in 12-month-old but not 3-month-old rats within 6-8 hr after APAP administration, perhaps secondary to declining renal function. After 500 mg/kg APAP, iv, blood APAP concentrations were markedly elevated in 12 month-old compared with 3-month-old rats during the entire course of the experiment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565203 TI - Effect of experimental diabetes on elimination kinetics of diflunisal in rats. AB - The effects of insulin-deficient diabetes on the elimination of diflunisal were investigated in streptozotocin-treated rats. Diflunisal, a fluorinated salicylate with nonsteroidal antiinflammatory properties, is eliminated primarily as the ester and ether glucuronides. After an iv injection of a 10 mg/kg dose, diabetic rats cleared diflunisal more rapidly than control rats; time-averaged total body clearances were 1.96 +/- 0.29 and 1.10 +/- 0.12 ml/min/kg, respectively. For a low clearance drug such as diflunisal, changes in the total body clearance can result from changes in the extent of plasma protein binding and/or drug metabolic rate. To determine whether the pronounced changes in elimination clearance in diabetic rats were due to the changes in plasma protein binding or enzyme activity, diflunisal was infused to obtain steady state kinetics. At steady state, the unbound intrinsic clearance increased from 43.4 +/- 16.4 ml/min/kg in the control rats to 82.5 +/- 21.1 ml/min/kg in diabetic rats at a high infusion rate (72 micrograms/min). When the infusion rate was lowered to 4.5 micrograms/min, the respective values for the unbound intrinsic clearance were 353 +/- 101 ml/min/kg and 561 +/- 112 ml/min/kg. Diabetic rats, however, showed no changes in plasma protein binding of diflunisal. The data suggest that the elimination of diflunisal was increased as a result of increased enzyme activity. Insulin treatment appeared to reverse the diabetic effect, suggesting that the effect on drug metabolism was the result of insulin deficiency and not a secondary or nonspecific effect of streptozotocin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565204 TI - Cocaine disposition in humans after intravenous injection, nasal insufflation (snorting), or smoking. AB - The disposition of radiolabeled cocaine in humans has been studied after three routes of administration: iv injection, nasal insufflation (ni, snorting), and smoke inhalation (si). Metabolism, followed by urinary excretion of metabolites, proved to be the major route of elimination in all cases. Hydrolytic products (benzoylecgonine, ecgonine methyl ester) were the major excretion products. Benzoyl ecgonine was generally most prevalent, but after smoking two subjects excreted larger amounts of ecgonine methyl ester and the ratio of the two compounds averaged lower in subjects who smoked cocaine. Low binding of cocaine to plasma proteins was observed and blood to plasma ratios were essentially unity. The volume of distribution of cocaine is low (2.70 liter/kg for V beta). Absorption of smoked cocaine was rapid (half-time of 1.1 min). Absorption after ni was slower (half-time of 11.7 min). After iv injection, a rapid distribution phase was observed (half-life of 11 min) and the elimination half-life was 78 min. In 16 subjects divided into three groups based on routes, the half-life based on the average rate constant was 69 min. Bioavailability was good after ni (80%). Undecomposed cocaine from si was well absorbed, but observed bioavailability was diminished by degradation from heating. PMID- 2565205 TI - On the antipyrine test in laboratory animals. Studies in the dog and monkey. AB - The antipyrine (AP) test has been challenged in species other than humans on the grounds that, in some nonhuman species, particularly on induction, hepatic blood flow may become as prominent a factor in AP clearance as hepatic metabolism. Therefore, we investigated in dogs and monkeys the disposition of AP to determine how well AP serves as a model drug to indicate changes in rates of hepatic clearance. After administration of an oral solution of AP (5 mg/kg) to control dogs, the percentage of the dose absorbed was 98%, based on urinary and fecal excretion of AP and its metabolites. Despite complete AP absorption, absolute bioavailability of AP was 78 +/- 12% under basal conditions, suggesting that AP does undergo some degree of presystemic elimination, approximately 22%. After PB administration of 20 mg/kg/day for 9 days, po, AP bioavailability decreased to 60 +/- 14%. The systemic clearance of AP increased from 9.4 +/- 2.3 ml/min/kg under basal conditions to 27.5 +/- 4.6 ml/min/kg following PB. PB decreased mean plasma AP half-life from 71.5 min under basal conditions to 27.7 min, and mean hepatic blood flow increased from 0.49 liters/min to 0.63 liters/min. Induction doubled the hepatic extraction ratio for AP to 0.4 from 0.2 under basal conditions. In beagle dogs after PB pretreatment, 97% of the total systemic clearance of AP was estimated to be due to enhanced hepatic AP metabolism, only 3% to increased hepatic blood flow. Therefore, for dogs under both basal and induced conditions it is concluded that AP clearance reflects predominantly hepatic AP metabolism, being negligibly influenced by hepatic blood flow.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565206 TI - The physiological disposition of lovastatin. AB - Lovastatin is a pro-drug lactone whose open chain beta-hydroxy-acid (HA) is a potent inhibitor of hydroxymethylglutaryl-CoA-reductase and thus of cholesterol synthesis. Because the liver is the major site of cholesterolgenesis, it is the principal target organ for agents of this class. In animals, lovastatin is not as well absorbed as HA given per se, but that fraction that is absorbed reaches the portal circulation largely unchanged and is more efficiently extracted by the liver, after which it is reversibly biotransformed to HA and irreversibly to other enzymatically active products. These, like HA, maintain high hepatic gradients relative to all tissues examined. The minimal systemic burden for HA is attributable in part to the metabolic equilibrium, lovastatin in equilibrium HA, the opposing reactions for which appear to be present in most tissues. Excretion is very largely biliary in all species. Detailed comparisons of absorption, distribution, metabolism, and excretion profiles presented here and elsewhere indicate dogs to be the most appropriate paradigm for humans for study of lovastatin disposition. PMID- 2565207 TI - Pharmacokinetics and metabolism of SK&F 86002 in male and female Sprague-Dawley rats. AB - Several pharmacokinetic parameters for SK&F 86002 [6-(4'-fluorophenyl)-5-(4' pyridyl)-2,3-dihydroimidazo(2,1-b)-thia zole] and its metabolites (sulfoxide, sulfone) were measured in male and female Sprague-Dawley rats after iv (5 mg/kg) and a wide range (10-80 mg/kg) of oral doses of SK&F 86002. In both sexes. SK&F 86002 is metabolized to an active metabolite, sulfone, which has an extended half life (approximately 13 hr) and, therefore, has the potential to accumulate upon repeated dosing. In addition, striking differences between sexes were noted in several of the pharmacokinetic parameters measured. The AUCs areas under the plasma concentration time curves, for SK&F 86002 in female rats obtained at the higher doses of SK&F 86002 were substantially greater than expected, when compared with lower doses in female rats or with equivalent doses in male rats. Furthermore, at all doses of SK&F 86002, AUCs for sulfoxide were substantially larger in female than in male rats. Consequently, the systemic exposure to SK&F 86002 and metabolites is substantially greater in female rats than in male rats. Therefore, extrapolation of the pharmacologic and toxicologic sequelae of SK&F 86002, observed at larger doses in female rats, to lower doses should be approached cautiously. Furthermore, the extended half-life of the pharmacologically active sulfone metabolite of SK&F 86002 suggests that this metabolite could accumulate on repeated daily dosing of SK&F 86002 and could, therefore, account for much of the chronic pharmacologic and toxicologic activity of SK&F 86002. PMID- 2565208 TI - N-methylation as a toxication route for xenobiotics. II. In vivo formation of N,N'-dimethyl-4,4'-bipyridyl ion (paraquat) from 4,4'-bipyridyl in the guinea pig. AB - The biotransformation of 4-phenylpyridine and 4,4'-bipyridyl to N-methylated quaternary ammonium metabolites in guinea pig and rabbit has been examined. Neither animal species excreted the neurotoxin N-methyl-4-phenylpyridinium ion as a urinary metabolite after ip administration of 4-phenylpyridine. However, treatment of rabbits with 4,4'-bipyridyl resulted in the formation of N-methyl 4,4'-bipyridinium ion in the urine (1.2% of the administered dose), and ip administration of 4,4'-bipyridyl to guinea pigs afforded both N-methyl-4,4' bipyridinium ion and N,N'-dimethyl-4,4'-bipyridinium ion (paraquat) as urinary metabolites (0.8% and 2.9%, respectively, of the administered dose). The detection of the lung toxin paraquat as a urinary metabolite of 4,4'-bipyridyl is a significant finding, in that it represents the first documented report of the formation of a toxic metabolite via the N-methylation pathway. PMID- 2565209 TI - Spironolactone metabolism in target tissues. Characteristics of deacetylation in kidney, liver, adrenal cortex, and testes. AB - Prior investigations demonstrated that many of the actions of spironolactone (SL) required deacetylation of the parent compound as the first step in the formation of biologically active metabolites. Studies were done to characterize the process of deacetylation in several target tissues. The reaction was catalyzed by microsomal and cytosolic fractions of livers, kidneys, adrenal glands, and testes. Microsomal activity was greatest in liver and kidney and far exceeded cytosolic metabolism in those tissues. In adrenal glands and testes, by contrast, deacetylation was greater in cytosolic than microsomal fractions. The metabolism mediated destruction of adrenal microsomal cytochromes P-450 by SL was enhanced by coincubation of microsomes with cytosol, illustrating the potential importance of combined microsomal and cytosolic metabolism in the actions of SL. The deacetylation of SL was decreased by various esterase inhibitors; the organophosphate compounds were the most potent inhibitors. The effectiveness of the esterase inhibitors varied from tissue to tissue, as well as from microsomes to cytosol within each tissue. The results indicate that SL deacetylation is catalyzed by microsomal and cytosolic esterases in various target tissues; several isozymes appear to be involved. These and prior observations suggest that tissue metabolism of SL is of major importance in the actions of the drug. PMID- 2565210 TI - Renal tubular secretion of amiloride and its inhibition by cimetidine in humans and in an animal model. AB - The histamine H2 antagonist cimetidine has been shown to reduce the renal tubular secretion of other organic cations through competition for the specific transport system with organic cations in the renal proximal tubule. The potential interaction between cimetidine and the potassium-sparing diuretic amiloride was investigated in humans and in the isolated perfused rat kidney. A chronic dosing study was conducted in eight healthy subjects who received, in random order, amiloride (5 mg daily), cimetidine (400 mg twice daily), both drugs together, and a control phase in which no drug was present. Cimetidine reduced the renal clearance of amiloride by a mean of 17%, from 358 +/- 134 to 299 +/- 118 ml/min (p less than 0.05), and the urinary excretion of amiloride from 65 +/- 11 to 53 +/- 13% of the dose (p less than 0.05). Amiloride reduced the excretion of cimetidine from 43 +/- 7 to 32 +/- 9% of the dose (p less than 0.05) and the area under the plasma concentration-time curve for cimetidine by a mean of 14% (p less than 0.05) but had no effect on the renal clearance of cimetidine. In the perfused rat kidney, cimetidine reduced the amiloride unbound renal clearance to glomerular filtration rate ratio from 5-7:1 to 1-2:1 (p less than 0.05). These studies demonstrate that cimetidine inhibits the renal tubular secretion of amiloride in humans and in rats to a similar extent. In addition, in humans the gastrointestinal absorption of both amiloride and cimetidine appear to be reduced by each other, by an as yet unknown mechanism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565211 TI - Metabolism of cyclosporin A. IV. Purification and identification of the rifampicin-inducible human liver cytochrome P-450 (cyclosporin A oxidase) as a product of P450IIIA gene subfamily. AB - A cytochrome P-450 involved in the metabolism of cyclosporin A (CsA) was isolated and purified to electrophoretic homogeneity from human liver microsomes of renal transplant donors. This cytochrome, designated P-450(CsA), exhibited a type I binding spectrum in the presence of CsA with a Ks(app) of 25 microM, a molecular weight of 52 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and a maximal absorbance at 449 nm when reduced in the presence of carbon monoxide. The N-terminal sequence of P-450(CsA), determined by Edman degradation reaction, was 63% homologous with that of the rabbit liver CsA oxidase P-450 3c and 100% homologous with that of the human liver isozyme P-450(HLp/NF), recently identified as the human nifedipine (NF) oxidase. Polyclonal and monoclonal antibodies directed against P-450 3c and P-450(HLp/NF), respectively, recognized native microsomal and highly purified P450(CsA). As observed in the rabbit, human liver microsomes were shown to generate mono- and dihydroxy, as well as dihydroxy and/or monohydroxy N-demethylated, derivatives of CsA. Production of these metabolites was shown to be specifically inhibited by anti-P-450 3c polyclonal antibodies. CsA oxidase, NF oxidase, and erythromycin demethylase were shown to be closely correlated with the level of P-450(CsA) determined from Western blot or enzyme-linked immunosorbent assay. Moreover, these monoxygenase activities and the hepatic level of P-450(CsA) were simultaneously increased in the liver of patients treated for 4 days with 600 mg of rifampicin per day. Finally, NF was shown to be a competitive inhibitor of CsA oxidation and vice versa. We conclude that P-450(CsA) is responsible for most (80%) of CsA oxidase activity in human liver, is encoded by gene P450IIIA3, as is NF oxidase, or a very closely related gene, and is strongly inducible by rifampicin pretreatment. PMID- 2565212 TI - The pharmacokinetics of albumin conjugates of D-penicillamine in rats. AB - A protein conjugate that forms during the metabolism of D-penicillamine (D-PEN) in rats is electrophoretically indistinguishable from the (D-PEN-albumin) mixed disulfide that forms during the oxidation of D-PEN in plasma in vitro. Both the conjugate formed in vivo and the mixed disulfide synthesized in vitro are eliminated slowly in rats, with half-lives of 5.33 +/- 0.25 days and 3.48 +/- 0.42 days, respectively. These half-lives approximate or exceed the half-lives of radioiodinated plasma protein in the animals. The stability of the D-PEN-albumin conjugate contrasts with the rapid elimination of D-PEN itself and probably explains delayed elimination of D-PEN-containing disulfides in humans. Stable modification of tissue proteins by D-PEN may also be involved in the mode of action of D-PEN in rheumatoid arthritis. PMID- 2565213 TI - Stereoselective metabolism and pharmacokinetics of racemic methylphenobarbital in humans. AB - The stereoselectivity of the metabolism and pharmacokinetics of methylphenobarbital (MPB) was studied in six healthy adult male volunteers given single oral doses of the racemic drug. All of the volunteers were phenotypically extensive metabolizers of the drug. The R- and S-enantiomers of MPB were analyzed in plasma by an enantioselective HPLC method, and the enantiomers of the 4 hydroxy-MPB metabolite in urine by a similar procedure. The (R)-MPB was extensively hydroxylated, with an average of 49.56% of that enantiomer being recovered in urine as (R)-4-hydroxy-MPB. Only 7.16% of the (S)-MPB was converted to the corresponding hydroxy metabolite. The extensive hydroxylation of (R)-MPB resulted in rapid elimination of this enantiomer, with a terminal plasma half life of 7.52 +/- 1.70 (SD) hr. The (S)-MPB, the only recognized metabolites of which were (S)-4-hydroxy-MPB and phenobarbital (PB), was eliminated very slowly [t1/2, 69.78 +/- 14.77 (SD) hr]. The oral clearance of (R)-MPB (0.470 +/- 0.184 (SD) liters/hr/kg) was much higher than that of (S)-MPB [0.017 +/- 0.001 (SD) liters/hr/kg]. The extreme differences in metabolic fate and pharmacokinetics of the enantiomers of MPB are interesting. Most of the circulating PB seemed to be derived from (S)-MPB. In other respects the pharmacodynamic implications of these pharmacokinetic differences are unclear, because the relative anticonvulsant potencies of the enantiomers of MPB are unknown. PMID- 2565214 TI - Metabolic interaction between morphine and naloxone in human liver. A common pathway of glucuronidation? AB - Metabolic interaction between naloxone and morphine has been studied in vitro in human liver microsomes. Morphine and naloxone are metabolized by an UDP glucuronyl transferase to their 3-glucuronides. There is a very good correlation between the rates of formation of morphine-3-glucuronide and of naloxone-3 glucuronide. Naloxone seems to have a 10-fold higher affinity for the enzyme binding site than does morphine. Morphine and naloxone show bisubstrate kinetic patterns which appear very similar. PMID- 2565215 TI - Species differences in protein binding of diflunisal. PMID- 2565216 TI - Tissue distribution of intravenously administered dimethyl sulfoxide in the normal rat. PMID- 2565217 TI - Hormonal control of tyrosine hydroxylase in the median eminence: demonstration of a central role for the pituitary gland. AB - In intact male rats the concentration of dopamine in hypophysial portal plasma of animals treated simultaneously with estradiol and progesterone was twice that of animals treated with the solvent vehicle. Treatment with estradiol or progesterone alone had no effect on dopamine in portal plasma. The rate of synthesis of dihydroxyphenylalanine (DOPA), the precursor of dopamine, in tuberoinfundibular dopaminergic (TID) neurites in the median eminence (ME) was 15 +/- 1.0 (mean +/- SE) pmol DOPA/ME.h in estradiol-progesterone-treated animals compared to 3.2 +/- 0.02 in vehicle-treated controls. Treatment with estradiol or progesterone alone gave a result similar to that seen in controls. In hypophysectomized animals treated with estradiol and progesterone, DOPA synthesis in the ME was greatly attenuated compared to that in intact rats. The in situ activity of tyrosine hydroxylase (TH; expressed as moles of DOPA per mol TH/h) in the ME was 178 +/- 16.5 in estradiol-progesterone-treated intact rats, but was 27 +/- 2.4, 52 +/- 4.2, and 35 +/- 2.5 in animals treated with the solvent vehicle, estradiol, and progesterone, respectively. In hypophysectomized rats the in situ activity of TH in the ME of animals treated with estradiol and progesterone was 53 +/- 8.4, which was significantly (P less than 0.01) less than that in similarly treated intact animals. The circulating PRL level in vehicle-treated animals was 35 +/- 4.6 ng/ml compared to 121 +/- 16 in estradiol-treated animals and 133 +/- 12.2 in estradiol- and progesterone-treated rats, indicating that the difference in the effects of estradiol and estradiol-progesterone on dopamine release, DOPA synthesis, and in situ TH activity was not solely due to a difference in circulating PRL levels. Maintenance for 7 days of anterior pituitary tissue as a graft in a lateral ventricle of intact rats resulted in a 2 fold increase in the synthesis of DOPA and TH activity in the ME compared to that in animals with liver implants. Results obtained in hypophysectomized animals with implants were similar to those in intact animals. The concentrations of PRL in cerebrospinal fluid of intact rats and hypophysectomized rats with anterior pituitary implants in the lateral ventricles were 96 +/- 32 and 127 +/- 35 ng/ml, respectively, which was significantly (P less than 0.001) greater than those in animals with liver implants. We suggest that a factor of pituitary origin stimulates TH activity in TID neurons. This stimulation may be due to PRL, but the existence of another stimulatory substance secreted by pituitary cells cannot be excluded. PMID- 2565219 TI - Regulation of pancreatic enzyme secretion in conscious rats by intraluminal somatostatin: mechanism of action. AB - These studies were undertaken to characterize 1) the immunoreactive somatostatin (SS) forms found in the gastrointestinal lumen and 2) to assess the possible role of luminal SS on pancreatic exocrine function. Results indicate that different forms of SS are secreted into the rat gastric and duodenal lumen in proportions corresponding to their gastric and duodenal contents. Inhibition of diversion stimulated pancreatic exocrine secretion by intraduodenal infusion of SS (SS-ID) is dose dependent with a maximal effective dose of 24 micrograms/kg-1h-1 for volume and 48 micrograms/kg-1h-1 for protein output. Infusion of supramaximal doses result in a loss of the inhibitory effect of SS-ID on both volume and protein outputs. Comparison between inhibition of pancreatic secretion by iv SS (SS-IV) and SS-ID indicates that SS-ID is about 20 times less potent than SS-IV in its inhibition of stimulated pancreatic exocrine secretion. Intraileal (SS-IL) infusion of SS at 48 micrograms/kg-1h-1 did not inhibit stimulated pancreatic secretion but was rather stimulatory possibly through the inhibition of putative ileal inhibitors. Likewise, passive immunization against circulating SS did not affect the inhibitory effects of SS-ID on pancreatic secretion. These data indicate that SS is secreted into the rat gastrointestinal lumen and that its infusion into the duodenal lumen inhibits stimulated pancreatic secretion in a dose-dependent fashion. The observation that SS-IL did not inhibit pancreatic secretion and that passive immunization against circulating SS did not affect the inhibitory effect of SS-ID suggest that the action of SS-ID on stimulated exocrine pancreatic secretion is probably indirect and may involve a duodenal signal, possibly an inhibition of endogenous release of cholecystokinin and/or secretin. PMID- 2565218 TI - Protein kinase C is not essential for growth hormone (GH)-releasing factor induced GH release from rat somatotrophs. AB - To examine the role of protein kinase-C in the mediation of GH release we used acutely dispersed purified somatotrophs in static incubation and acutely dispersed adenohypophyses in perifusion. In static incubation, activation of protein kinase-C by phorbol 12-myristate 13-acetate (PMA) and 1,2-dioctanoyl-rac glycerol (diC8) resulted in an increase in GH release and a concurrent concentration-dependent increase in cAMP accumulation. The GH response to diC8 in perifusion was reversible and repeatable. On the other hand, the GH response to PMA was not repeatable. The lack of repeatability is most likely due to the depletion of protein kinase-C by prolonged treatment with PMA. This assumption is strengthened by the observation that 1 h of perifusion with PMA left the somatotrophs refractory to a subsequent application of diC8. When graded pulses of GRF were applied during treatment with PMA, the GH response to GRF was not altered. Somatostatin reduced (in static incubation) or blocked (in perifusion) the release of GH induced by diC8 and PMA, but the accumulation of cAMP was not affected. We conclude that 1) activation of protein kinase-C in normal somatotrophs results in GH release which may not be completely independent of the cAMP pathway; 2) activation of protein kinase-C is not essential for GRF-induced GH release; and 3) SRIF acts at a site distal to or independent of cAMP to inhibit GH release induced by activators of protein kinase-C. PMID- 2565220 TI - Involvement of the hippocampus in central nervous system-mediated glucoregulation in rats. AB - To find out whether the hippocampus is involved in central nervous system mediated glucoregulation, we injected saline, neostigmine, dopamine, norepinephrine, bombesin, beta-endorphin, somatostatin, and prostaglandin F2 alpha into the dorsal hippocampus in anesthetized fed rats. After injection of dopamine, norepinephrine, bombesin, beta-endorphin, somatostatin, or prostaglandin F2 alpha, the level of hepatic venous plasma glucose did not differ from that in saline-treated control rats. However, neostigmine, an inhibitor of acetylcholine esterase, caused a dose-dependent increase in the hepatic venous plasma glucose concentration. This neostigmine-induced hyperglycemia was dose dependently suppressed by coadministration of atropine, but not by hexamethonium. Injection of neostigmine (5 X 10(-8) mol) resulted in an increase not only in glucose but also in glucagon, epinephrine, and norepinephrine in hepatic venous plasma. In bilateral adrenalectomized rats, neostigmine-induced hyperglycemia was suppressed, but the hepatic venous plasma glucose concentration still increased significantly. These results indicate that the hippocampus is involved in central nervous system-mediated glucoregulation through cholinergic muscarinic activation, partly via epinephrine secretion. PMID- 2565221 TI - Evidence that gonadotropin-releasing hormone also functions as a growth hormone releasing factor in the goldfish. AB - The present study examined the influence of GnRH on the in vivo and in vitro secretion of GH in the goldfish (Carassius auratus). Intraperitoneal injection of several GnRH peptides, including a form native to goldfish, salmon GnRH (sGnRH), elevated circulating GH levels in female goldfish. An analog of mammalian GnRH (mGnRH), [D-Ala6,Pro9-NEt] mGnRH (mGnRH-A), at a dosage of 0.1 microgram/g BW increased serum GH levels for up to 48 h after a single ip injection. Goldfish receiving a series of injections of this dose of mGnRH-A also displayed an increased rate of body growth, indicating that the mGnRH-A-induced increase in the circulating GH level was sufficient to accelerate body growth. In vitro experiments using perifused pituitary fragments found that sGnRH stimulated the secretion of GH from the goldfish pituitary in a potent, dose-dependent, and reversible manner. The time course of response and half-maximally effective dose of sGnRH were very similar for both GH and gonadotropin (GTH) secretion in vitro, suggesting that the mechanism(s) mediating the stimulatory actions of GnRH in the goldfish may be similar for both GH and GTH secretion. However, GnRH-induced GH and GTH secretion from the goldfish pituitary can occur independently of each other, as demonstrated by the finding that somatostatin inhibited the GnRH stimulation of GH secretion in vitro, without influencing the GTH response, whereas the dopamine agonist apomorphine inhibited GnRH-induced GTH secretion in vitro, without influencing the GH response. Furthermore, the dopamine antagonist pimozide did not influence serum GH levels, although pimozide potentiated the stimulatory effect of GnRH on GTH secretion in vivo by blocking the endogenous GTH release inhibitory action of dopamine. Results of the present study suggest that the secretion of GH and GTH in the goldfish are regulated, at least in part, through a common releasing factor, GnRH, whereas somatostatin and dopamine appear to act independently as GH and GTH release inhibitory factors, respectively. PMID- 2565222 TI - Failure of non-surgical procedures to treat gastric trichobezoar. PMID- 2565223 TI - Esophageal varices: current therapy in 1989. PMID- 2565224 TI - Variation in some enzymes in amniotic fluid and maternal serum during pregnancy. AB - The following 10 enzymes were assayed in 187 amniotic fluid and maternal serum samples at 15-42 weeks of gestation: alkaline phosphatase, heat-stable alkaline phosphatase (only in amniotic fluid), acid phosphatase, alanine aminotransferase, aspartate aminotransferase, alpha-amylase, gamma-glutamyltransferase, creatine kinase, lactate dehydrogenase, and lysozyme. The normal reference ranges are reported for amniotic fluid and maternal serum enzymes, together with the abnormal values accompanying neural tube defects and EPH-gestosis. The determination of gamma-glutamyltransferase, heat-stable alkaline phosphatase and creatine kinase was found to be of appreciable diagnostic significance in clinical practice. PMID- 2565225 TI - Enzyme levels in bronchoalveolar lavage fluid and serum of active pulmonary tuberculosis patients. AB - Bronchoalveolar lavage (BALF) was found to be a useful index of cell damage. It has been observed that the enzymes in BALF could give an idea of cell damage in a pulmonary disease. Acid phosphatase, alkaline phosphatase, leucine aminopeptidase and gamma-glutamyl-transpeptidase were assessed in mild, moderate, and severe pulmonary tuberculosis patients and Mantoux-negative normal controls. Activities of these enzymes were found to be higher in patients and were increasing with the severity of the disease. Increase in these enzymes in pulmonary tuberculosis patients could be attributed to lung tissue damage. PMID- 2565226 TI - Activity of the gamma-glutamyltransferase, leucine aminopeptidase and alkaline phosphatase enzymes in human tear fluid. AB - In this work the presence of gamma-glutamyltransferase (GGT), leucine amino peptidase (LAP) and alkaline phosphatase (AP) is shown in human tear fluid. We studied these levels according to sex, age and some eye refraction defects. The differences between the levels for both sexes are not significant. LAP and AP do not show any differences in either age groups or individuals with some refraction defects. The average level of GGT is higher from 40 years of age upwards (p less than 0.005). In individuals with refraction defects, the enzymatic activity is significantly higher (p less than 0.05) than the activities found in normal subjects. The levels of the three enzymes in serum and tear fluid do not show a significant correlation nor are they significantly modified after the samples have been frozen for a month at -20 degrees C. PMID- 2565227 TI - Life stress and lymphocyte alterations among patients with rheumatoid arthritis. AB - The relation between life stress and immune parameters was investigated for 33 female rheumatoid arthritis (RA) patients interviewed during three routine monthly clinic checkups. Life stress from major and minor events, coping efficacy, and self-reported psychological distress were assessed, and immunofluorescence of T-cells and B-cells was performed on the blood drawn during each visit. Small stressful events were positively related to the proportion of circulating B-cells, psychological distress was inversely related to proportion of circulating T-cells, and major life events were associated with lower T helper/T-suppressor cell ratios. PMID- 2565228 TI - Androgen and oestrogen response to a single injection of hCG in cryptorchid horses. AB - Androgen (testosterone and androstenedione) and oestrogen (oestradiol -17 beta and oestrone) concentrations were measured by radio-immunoassay in the peripheral plasma of two geldings (five-years-old), three bilateral cryptorchids (two, two and a half, and five-years-old) and three normal intact stallions (four, five and five and a half-years-old) before and after a single injection of 10,000 iu human chorionic gonadotrophin (hCG). In the stallions, hCG administration resulted in an immediate sharp increase of conjugated oestrogens and a more gradual increase of unconjugated androgens. In the cryptorchids, the unconjugated androgens increased following a similar pattern to that observed in the stallions, but reached lower peak values, whereas the conjugated oestrogens showed only a very slight increase. In the stallions and cryptorchids, the maximum oestrogen levels were reached two days after injection, whereas the maximal levels for androgens were reached a day later. In the geldings, hCG injection had no effect on plasma steroid levels. It is suggested that the measurement of unconjugated androgens (testosterone or/and androstenedione) before and three days after intravenous injection of 10,000 iu hCG may prove useful for the diagnosis of cryptorchidism or exploration of testicular function in stallions. PMID- 2565229 TI - Blockade of sustained repetitive action potentials in cultured spinal cord neurons by zonisamide (AD 810, CI 912), a novel anticonvulsant. AB - Zonisamide is a novel anticonvulsant that prevents seizures in laboratory animals and in man. Zonisamide (3 micrograms/ml and above) blocked the sustained firing of action potentials induced by depolarizing steps of current injected across the membrane of intracellularly recorded spinal cord neurons. Responses to GABA and glutamate were not altered by zonisamide, and spontaneous synaptically evoked activity was not reduced until higher concentrations of zonisamide (10 micrograms/ml) were applied. PMID- 2565230 TI - Clorazepate in dogs: tolerance to the anticonvulsant effect and signs of physical dependence. AB - Dogs were treated with clorazepate, which is known to be completely metabolized to desmethyldiazepam. 2 mg/kg t.i.d. were given orally for 5-6 weeks, a dose regimen providing plasma concentrations of desmethyldiazepam in the range known to be therapeutic in man. The rate of development of tolerance to the anticonvulsant effect was followed by weekly determinations of the convulsive threshold for pentetrazole before, during and after cessation of treatment. The development of tolerance was not as clear and pronounced as that found after treatment with diazepam and clonazepam in earlier studies with dogs. The seizure threshold was elevated by a factor of 1.2-3.5 during the first 2 weeks of treatment; during the following weeks, tolerance developed in only 2 out of 6 dogs (decline of the pentetrazole threshold in spite of rising or unchanged plasma concentrations). 36 h after withdrawal of clorazepate, the convulsive threshold had fallen below the control values in all dogs, but 1 week later it had returned to the control level. One day after cessation of treatment, 2 out of 6 dogs showed withdrawal seizures, which, in 1 case, were lethal. This shows that severe withdrawal symptoms, even lethal seizures, may appear after abrupt discontinuation of chronic clorazepate treatment, in spite of the relatively low tolerance liability of clorazepate. PMID- 2565231 TI - The kinetics of the thermal deactivation of transglutaminase from guinea-pig liver. AB - By incubating native (N) transglutaminase from guinea-pig liver at various temperatures and assaying it at 25 degrees C, two steps in the irreversible deactivation process to the denatured form (D) have been found. The fitting of the data to the equations of two possible models (the two-steps model and the two isoenzymes model) is only compatible with the first one (N----X----D). It is shown that the structure of the active intermediate, X, depends on the deactivation temperature and on the thermal history of the enzyme. This may mean that transglutaminase exists in a large number of microstates. Surprisingly, the activation energy of deactivation is lower than that of activity (36.6 +/- 3.4 against 47.2 +/- 2.2 kJ.mol-1). By deactivating transglutaminase at a constant temperature (55 degrees C) and assaying it at variable temperatures, the activation energy of the intermediate, (X55), has been determined to be 40.2 +/- 5 kJ.mol-1, of the same order of magnitude as the native form. Among several agents assayed, only Ca2+ had a positive effect on the thermal stability of this enzyme. At 40 degrees C, transglutaminase was quite stable in the presence of Ca2+ (in its absence, the half-life was 65 min) and at 45 degrees C, its thermostability had been considerably increased, the half-life being raised from 47 min to 275 min. PMID- 2565232 TI - Properties of D-amino-acid oxidase from Rhodotorula gracilis. AB - The flavoprotein D-amino-acid oxidase was purified to homogeneity from the yeast Rhodotorula gracilis by a highly reproducible procedure. The amino acid composition of the protein was determined; the protein monomer had a molecular mass of 39 kDa and contained one molecule of FAD. The ratio between A274/A455 was about 8.2. D-Amino-acid oxidase from yeast showed typical flavin spectral perturbations on binding of the competitive inhibitor benzoate and was reduced by D-alanine under anaerobiosis. The enzyme reacted readily with sulfite to form a covalent reversible adduct and stabilized the red anionic form of the flavin semiquinone on photoreduction in the presence of 5-deazariboflavin; the 3,4 dihydro-FAD form was not detectable after reduction with sodium borohydride. Thus D-amino-acid oxidase from yeast exhibited most of the general properties of the dehydrogenase/oxidase class of flavoproteins; at the same time, the enzyme showed some peculiar features with respect to the same protein from pig kidney. PMID- 2565233 TI - The acute effects of intravenous xamoterol ('Corwin', I.C.I. 118, 587) on resting and exercise haemodynamics in patients with mild to moderate heart failure. AB - The known properties of xamoterol, a partial beta 1-agonist, provide a basis to pharmacologically modulate cardiac responses to variations in sympathetic tone. Haemodynamic variables were assessed at rest and on exercise before and after intravenous xamoterol (0.2 mg kg-1), in 30 patients with mild to moderate cardiac failure. Xamoterol produced significant improvements in resting cardiac index (2.51 +/- 0.15 to 2.80 +/- 0.14 l min-1 m-2; P less than 0.001), stroke volume (62 +/- 4 to 75 +/- 5 mljbeat-1; P less than 0.001) and stroke work index (42.4 +/- 3.6 to 47.7 +/- 3.9 gm beat-1 m-2; P less than 0.01). This occurred despite a significant reduction in heart rate (78 +/- 3 to 74 +/- 2 beats min-1; P less than 0.05). There were also significant reductions in systemic vascular resistance (1990 +/- 141 to 1669 +/- 112 dynes s-1 cm-5; P less than 0.01) and double product (1146 +/- 46 to 1051 +/- 41 mmHg min-1 x 10(-1); P less than 0.05), with no significant changes in systolic blood pressure, pulmonary wedge pressure or ejection fraction. Xamoterol significantly attenuated the heart rate response to exercise (112 +/- 4 to 97 +/- 3 beats min-1; P less than 0.001), with no impairment in the expected exercise induced increase in cardiac index. This was due to the significant increase in stroke volume from 81 +/- 6 to 95 +/- 7 ml beat-1 (P less than 0.001). There were no significant changes in resting or exercise noradrenaline levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565234 TI - The future of saphenous vein as a coronary artery bypass conduit. AB - Continuing widespread use of autologous saphenous vein for coronary artery bypass grafting seems unavoidable despite its poor-term patency. We review here the evidence that platelet activation is responsible for early and late vein graft occlusion and conclude that other mechanisms probably contribute to late occlusions. We suggest that a rational strategy to improve vein graft patency should include: improved endothelial preservation during surgical implantation; use of better antiplatelet agents, in particular those which prevent platelet adhesion as well as aggregation; reduction of risk factors including serum cholesterol; and application of agents (e.g. heparin) which inhibit smooth muscle cell proliferation directly. We draw parallels between the pathogenesis of vein graft occlusion and coronary atherosclerosis and suggest that testing strategies for improving vein graft patency may also shed light on atherogenesis. PMID- 2565235 TI - Analysis of gene dosage on chromosome 11 in children suffering from Beckwith Wiedemann syndrome. AB - The Beckwith-Wiedemann syndrome (BWS) is composed of multiple congenital malformations coupled with a high concurrent risk for the development of specific rare childhood tumours. The syndrome is characterised by a complex mode of inheritance, but recent evidence indicates that it is an autosomal dominant trait with variable penetrance. It has been previously suggested that major rearrangements of the short arm of chromosome 11 are involved in the aetiology of the disease. We undertook to search for rearrangements in 11p in four patients with BWS and their parents and siblings. By using cloned DNA fragments homologous to four genes located on 11p, namely catalase, parathyroid hormone, insulin-like growth factor II and the proto-oncogene c-Ha-Ras, we subjected DNA from the patients to a restriction fragment length polymorphism (RFLP) analysis after digestion with restriction enzymes. We found no evidence for any large scale deletions or amplifications in this chromosomal region. We therefore conclude that altered gene dosage is not, as has been suggested, a requirement for the development of BWS. This raises the question of whether some other molecular mechanism is responsible for the malformations observed. PMID- 2565236 TI - Isoelectric focusing of bovine major histocompatibility complex class II molecules. AB - Serological approaches have been relatively unsuccessful in defining the allelic products of the bovine major histocompatibility (MHC) class II loci. We demonstrate that bovine class II allelic products can be characterized by precipitation with a polyclonal antiserum and separation using one-dimensional isoelectric focusing. Polymorphic beta chains were present in immunoprecipitates from both biosynthetically and surface-labeled lectin-stimulated bovine T cells. Precipitates from biosynthetically labeled but not surface-labeled T cells contained a basic invariant chain and a non-polymorphic structure. The non polymorphic structure appears to be a beta chain. The polymorphic class II beta chain co-segregated with bovine MHC class I allelic products in a half-sibling family, providing evidence for linkage between bovine class I and class II loci. This approach to the biochemical analysis of the bovine class II structures should facilitate the investigation of the association between the bovine products and disease susceptibility. PMID- 2565237 TI - Dynorphin stimulates the release of ANP from isolated rat atria. PMID- 2565238 TI - Biochemical and pharmacological characterization of ICI 198,615: a peptide leukotriene receptor antagonist. AB - ICI 198,615 is one representative of a new chemical class of peptide leukotriene receptor antagonists that are the most potent and selective described to date. ICI 198,615 antagonized LTC4-, LTD4- and LTE4-induced increases in cutaneous vascular permeability in the guinea pig, with i.v. ED50 values of 0.083, 0.11 and 0.067 mumol/kg, respectively. Against LTD4, ICI 198,615 was 615 and 415 times more potent than LY 171883 and FPL 55712, respectively. L-Serine borate, an inhibitor of the metabolism of LTC4 to LTD4, did not influence the antagonism by ICI 198,615 of LTC4-induced increases in cutaneous vascular permeability. The compound both inhibited and reversed aerosol ovalbumin antigen-induced increases in pulmonary resistance in passively sensitized guinea pigs, but demonstrated little ability to inhibit or reverse ovalbumin antigen-induced decreases in dynamic lung compliance. At concentrations ranging from 10(-8) to 10(-5) M, ICI 198,615 had no significant effect on either the spontaneous or ovalbumin antigen induced release of histamine, peptide leukotrienes, thromboxane B2 or 6-keto prostaglandin F1 alpha from chopped guinea pig lung. At 10 microM, the compound did not inhibit 5-, 12- or 15-lipoxygenase. Finally, ICI 198,615 antagonized LTD4 induced increases in TxB2 release from chopped guinea-pig lung. PMID- 2565239 TI - Nebivolol is devoid of intrinsic sympathomimetic activity. AB - Nebivolol is a chemically novel, potent and selective beta 1-adrenoceptor blocking agent that acutely lowers arterial blood pressure in hypertensive patients and rats without depressing, or even enhancing, left ventricular function. These properties could be compatible with a partial agonistic effect of beta-adrenoceptor-blocking agents. It was the aim of the present study to investigate whether nebivolol has intrinsic sympathomimetic properties. The study was performed on reserpinized dogs and spontaneously hypertensive rats, and on various isolated tissues from various species. Unlike pindolol and practolol, nebivolol did not exert a stimulating effect on the heart rate and left ventricular function in reserpinized animals and/or in isolated atria of reserpinized rats at doses that are clinically active. Nebivolol did not induce relaxation of isolated coronary arteries and saphenous veins at concentrations that block beta-adrenoceptors. These findings indicate that nebivolol is devoid of intrinsic sympathomimetic activity at clinically relevant doses. PMID- 2565240 TI - Evaluation of delta receptor mediation of supraspinal opioid analgesia by in vivo protection against the beta-funaltrexamine antagonist effect. AB - The involvement of delta opioid receptors in supraspinal analgesia was investigated. With this aim, opioids that produced analgesia in the tail immersion test were administered i.c.v. to mice a few minutes before the irreversible antagonist, beta-funaltrexamine (beta-FNA). Protection of the respective analgesic effects from beta-FNA blockade was obtained when evaluated 24 h later. Moreover, mu ligands protected the analgesia evoked by ED50s of morphine, [D-Ala2,N-Me-Phe4,Met-(o)5-ol]enkephalin (FK 33-824), [D-Ala2,N-Me Phe4,Gly-ol5]enkephalin (DAGO) and human beta-endorphin at doses (ED50s) lower than those required for delta ligands (approximately ED90s) to reach a similar protection. delta Preferential ligands effectively protected the analgesia induced by ED50s of [D-Ala2,D-Leu5]enkephalin (DADLE), [D-Thr2,Leu5]enkephalin Thr6 (DTLET) and [D-Pen2,D-Pen5]enkephalin (DPDPE) from the beta-FNA deteriorating effect. FK 33-824 and DAGO also provided good protection of the analgesia elicited by these delta ligands whereas morphine protected much less. Binding studies after i.c.v. injection of beta-FNA showed that its alkylating effect on opioid receptors was restricted to periventricular areas. In PAG, where the mu/delta receptor ratio is about 10, [3H]DADLE specific binding was still present after ED50s of DPDPE, DAGO, morphine and DADLE as protecting agents. [3H]Dihydromorphine [( 3H]DHM) binding was well protected by ED90s of morphine and DAGO, and to a lesser extent by DPDPE and DADLE. These results suggest that delta ligands, after binding to delta receptors, also need to act upon mu receptors to produce high levels of supraspinal analgesia in the tail immersion test. PMID- 2565241 TI - Effects of human ANF-(99-126) on sympathetic nerve function, hemodynamic parameters and plasma cGMP levels in anesthetized, ganglion-blocked dogs. AB - The ability of human ANF-(99-126) (ANF) to affect neuronal function, plasma cGMP levels or various hemodynamic parameters was tested in anesthetized dogs subjected to ganglionic blockade which prevented reflex changes in sympathetic tone. In each group of dogs, heart rate changes due to electrical stimulation of the cardioaccelerator nerve at 0.5, 1, 3 and 10 Hz were measured before and after ANF or vehicle infusion. A possible direct effect of ANF on the beta-adrenoceptor mechanisms was assessed by evaluating changes in heart rate after i.v. administration of isoproterenol (a beta-adrenoceptor agonist). The results showed that i.v. infusion of ANF (200 pmol/kg per min for 60 min), but not vehicle, produced significant inhibition of heart rate responses to low frequency nerve stimulation (0.5-1 Hz) when compared to vehicle 30 min after the end of infusion; high frequency (10 Hz) responses remained unaffected. A significant (P less than 0.05) enhancement in isoproterenol responses was evident in ANF-treated animals at the end of infusion, and 30 min afterward. ANF increased plasma cGMP from pretreatment levels of 28 +/- 4 to 149 +/- 1 pmol/ml at the end of infusion, and falling to 63 +/- 7 pmol/ml 30 min later (t 1/2 = 25.1 +/- 2.2 min). Cardiac output also tended to fall during infusion and remained significantly reduced ( 18%) 30 min after infusion's end; blood pressure fell without a decrease in total peripheral resistance. Thus, ANF produced a direct, frequency-dependent depressant action on sympathetic nerve function, reduced both cardiac output and blood pressure, and enhanced chronotropic responses to beta-adrenoceptor stimulation. These persistent changes in hemodynamic parameters and neuronal function may contribute to the mechanism of action of ANF. PMID- 2565243 TI - Spinal dynorphin involvement in the analgesia of pregnancy: effects of intrathecal dynorphin antisera. AB - In both rats and humans there is an analgesia associated with pregnancy. This analgesia is spinally mediated and involves the kappa type of opiate receptor. The current study demonstrates that intrathecal administration of high affinity dynorphin antibodies produces a significant reduction in jump thresholds during pregnancy (day 20). The administration of pre-adsorbed antisera fails to produce this effect. These results support the hypothesis that a spinal dynorphin/kappa opiate receptor system is activated during gestation. PMID- 2565242 TI - [3H]idazoxan binding at non-alpha 2-adrenoceptors in rabbit adipocyte membranes. AB - The imidazoline ligand, [3H]idazoxan, labels a large population of high-affinity binding sites in rabbit fat cell membranes (Bmax = 1370 +/- 91 fmol/mg protein; KD = 1.6 +/- 0.6 nM) when imidazoline derivatives are used for definition of non specific binding. [3H]Idazoxan sites are not alpha 2-adrenoceptors as assessed by competition studies which showed that epinephrine, norepinephrine and yohimbine do not inhibit [3H]idazoxan binding. Naphazoline, tramazoline and the Na+/H+ exchange inhibitor, amiloride, completely inhibited [3H]idazoxan binding. The Ki values were 9, 27 and 48 nM, respectively. PMID- 2565244 TI - Effects of neuraminidase on the regulation of erythropoiesis: III: Characterization of carbohydrate moieties on the surface of thymic regulatory cells that interact with erythroid colony-forming cells. AB - In this study we further define cell surface carbohydrate structures relevant to cellular interactions that regulate erythropoiesis. An analysis of thymocyte cell surface negativity was made using fluoresceinated poly-L-ornithine (FITC poly-L ornithine) as a probe that binds to negatively charged sites (i.e., sialic acid residues) at the cell surface. Two distinct subpopulations are labeled, comprising both intensely as well as weakly fluorescent subpopulations of thymocytes. Prior treatment of thymocytes with Vibrio cholerae neuraminidase (VCN), which removes cell surface sialic acid residues, markedly reduced the FITC poly-L-ornithine surface labeling of these cells. Distinct enzymatic modifications of regulatory cell functions were also assessed by the ability of thymocytes to function as separate regulatory subpopulations. Confirming our previous observations, treating thymocytes with VCN impaired the enhancement activity but had little effect on thymocyte regulatory ability to suppress erythroid colony growth. In contrast, treatment of thymocytes with galactose oxidase (GAO) or beta-galactosidase (beta-GAL) removed suppressor activity either before or after VCN treatment. A further exposure of GAO-treated thymocytes to sodium borohydride or hydroxylamine, which reduce D-galactose residues, restores their suppressor function and prevents enhancement. These differential enzymatic effects on thymocyte regulatory cell functions suggest that different carbohydrate structures may be involved in helper and suppressor activities for erythroid colony formation. Sialic acid residues may be associated with certain cells that function to enhance erythropoiesis, and D-galactose residues may be associated with the suppressor subpopulation. PMID- 2565245 TI - Evidence for the occurrence of an enkephalin-like peptide in adrenaline and noradrenaline neurons of the rat medulla oblongata. AB - The indirect immunofluorescence technique was used to analyze the catecholaminergic neurons in the medulla oblongata of the rat for the presence of enkephalin (ENK)- and neuropeptide Y (NPY)-like immunoreactivity (LI). In colchicine pretreated animals, using a double staining technique with mouse and rabbit antibodies against ENK and tyrosine hydroxylase (TH) or phenylethanolamine N-methyltransferase (PNMT), it was demonstrated that both TH- and ENK-LI occurred in the same neurons, particularly in many neurons of the A1 noradrenaline cell group. In the transition zone to the C1 adrenaline cell group, a proportion of PNMT-positive cells also contained ENK-LI. In the rostral and mid portion of the C1 group only few TH/PNMT-positive cells were found to be ENK-positive. In the noradrenergic A2 region, a moderate number of cell bodies also contained TH- plus ENK-LI, whereas only a few of the adrenaline cells of the C2 and C3 groups showed ENK-LI. In addition, with an elution restaining technique it was possible to demonstrate that several of the cells containing TH- and ENK-LI were also positive for NPY-LI. The present findings demonstrate that a subpopulation of the catecholaminergic neurons in the medulla oblongata of the rat is ENK-positive, thereby indicating a possible co-release of the two compounds in their projection areas, for example the paraventricular nucleus and the spinal cord. PMID- 2565246 TI - Role of brain acetylcholine in vasopressin release during osmotic stimulation and hemorrhage. AB - There is considerable evidence to suggest that there is a cholinergic link in the neural control of vasopressin release, but the precise role for this link has not been adequately demonstrated in the intact animal. We have, therefore, examined in conscious unrestrained rats the effects of central cholinergic blockade on the stimulation of vasopressin release by increased plasma osmotality (iv infusion of 2.5 M NaCl at 0.1 mg/kg body weight.min for 30 min) and by decreased blood volume (2 successive hemorrhages of 10% of blood volume each). The vasopressin responses to these stimuli were unaffected by either intracerebroventricular (icv) atropine (10 micrograms; muscarinic blockade) or icv hexamethonium (10 micrograms; nicotinic blockade) in doses which block the vasopressin responses to icv cholinergic agonists. The implications of these findings are discussed. PMID- 2565247 TI - Chronic intrastriatal L-pyroglutamate: neuropathology and neuron sparing like Huntington's disease. AB - Continuous injection of L-pyroglutamate (L-PGA) into the rat striatum induces a lesion with three regions: a necrotic core, a rim of pyknotic cells, and a peripheral spongiose region. The L-PGA was administered through an implanted intrastriatal cannula coupled to an Alzet osmotic pump loaded with one of three doses of L-PGA (3, 5, or 13 times the normal amount of L-PGA/g wet wt rat forebrain (23 nmol/g). The magnitude of the lesion was dependent upon the concentration of buffered L-PGA and the duration of continuous pumping. The necrotic region contained macrophages and neutrophils, while condensed neurons and oligodendroglial cells were present in the pyknotic region. The spongiose region contained vacuolated neuropil and degenerating nerve cells and oligodendroglia. The spongiose region blends with the normal neuropil. A population of aspiny neurons were identified throughout the spongiose region. These neurons stained positive for NADPH diaphorase and demonstrated a somatostatin-like immunoreactivity similar to that of the aspiny neurons spared in Huntington's disease and in the neurotoxin-induced striatal-lesioned rat models of Huntington's disease. PMID- 2565248 TI - Latent iron deficiency alters gamma-aminobutyric acid and glutamate metabolism in rat brain. AB - A diet containing 18-20 mg iron/kg to young weaned rats for 8 weeks altered the metabolism of gamma-aminobutyric acid and glutamate in the central nervous system without affecting blood hemoglobin. Subsequent rehabilitation with 390 mg iron/kg diet for 2 weeks normalized these changes. PMID- 2565249 TI - [Intermediator relation of the CNS and the improvement in the agents for therapy of drug poisoning]. AB - Intoxication induced by any "synaptic" poison is followed by the involvement in the pathological process of not only the mediator "target-system" but also other closely interrelated neurotransmitter systems of the brain. The functional condition of every of the systems may be of certain importance for the development of evident manifestations of intoxications. The study of the laws of the interactions of neurotransmitter systems in the central nervous system and their responses to intoxications may become the basis for the search of new effective drugs enhancing efficacy of classical antidotes of synaptic poisons. PMID- 2565250 TI - [Effect of salazosulfapyridine (sulfasalazine) on the function of the complement system in rats with adjuvant arthritis]. AB - In experiments on 24 albino male rats weighing 150-200 g it was found that the complement system and autoimmune humoral reactions are involved in the development of adjuvant arthritis. The use of sulfasalazine since the 10th day after intragastric administration of Freund complete adjuvant in a daily dose of 250 mg/kg for 2 weeks increased the activity of the complement system and decreased the autoimmune component. The data obtained clarify one of the possible mechanisms of allergic reactions to sulfasalazine which is related to the complement system activation. PMID- 2565252 TI - Prosomes discriminate between mRNA of adenovirus-infected and uninfected HeLa cells. AB - Prosomes are small cytoplasmic RNP complexes associated with repressed mRNA. In in vitro translation, they discriminate between the mRNA of adenovirus-infected HeLa cells and those of uninfected cells grown under normal conditions. Prosomes as well as their RNA constituents interact much more strongly with poly(A)+ mRNA of infected cells and inhibit their translation in vitro preferentially. A possible role of prosomes in the differential regulation of translation is discussed. PMID- 2565253 TI - Ethics and human values in family planning. PMID- 2565251 TI - [Comparative evaluation of the effect of carbidine and levamisole on the E rosette-forming activity of the lymphocytes in alcoholic intoxication]. AB - The E-rosette formation activity of lymphocytes in human alcoholics treated with carbidine was studied. In experimental alcohol intoxication in animals the effect of carbidine on the E-rosette formation was compared with that of levamisole. The effects of the both drugs in the in vitro experiments were studied as well. The use of carbidine for treating alcoholism was shown to produce changes in the E rosette formation indicating possible immunodulating properties of the drug. In experimental conditions of the immune response to ram erythrocytes during alcoholic intoxication the inhibitory effect of carbidine on the E-rosette formation was more pronounced. In the in vitro conditions carbidine changes the E rosette formation in alcoholics similarly to levamisole suggesting the immunomodulating properties of carbidine. PMID- 2565254 TI - The stability and short-term fluctuation in serum oxytocinase activity in pregnancy. AB - Maternal serum samples obtained from 123 normal pregnant women between 28 and 40 weeks gestation were analysed for oxytocinase (EC 3.4.11.3) activity using s benzyl-1-cysteine-4-nitroanilide as substrate. Oxytocinase activity was found to be stable at varying temperatures, with a mean co-efficient of variation of 5.0%. The short-term fluctuation of oxytocinase activity had a mean coefficient of variation of 3.6%. There is no suggestion of a "pulsatile" release of oxytocinae from its placental origin. PMID- 2565255 TI - Cesarean sections for maternal indications in Kasongo (Zaire). AB - Cesarean section rates in a rural area in Eastern Zaire are described. Indications are stringent and section is essentially performed on maternal indications. In the urban area cesarean section was performed for 1.1% of the expected births over a 9-year period, but only for 0.3% of expected births in the rural area, indicating a major problem in access to the district hospital. Section rates are compared with those reported for other settings. PMID- 2565256 TI - The intramural segment and the uterotubal junction: an anatomic and histologic study. AB - The injected latex method was used to study the anatomy of the intramural segment in ten uteri. Another ten uteri were used for its histological evaluation. The intramural segment had an S-shaped course with double curves. Its length ranged between 0.9 and 1.7 cm. The lining epithelium showed variations at different levels with increase in cell density and number of cilia as the isthmus was approached. At the uterotubal junction, there were mucosal folds of endosalpinx directed towards the tubal lumen. PMID- 2565257 TI - The regional versus national incidence of ectopic pregnancy in Finland from 1966 to 1986. AB - The annual incidence of ectopic pregnancy (EP) per deliveries, per all diagnosed pregnancies and per female population at fertile age (15-44 years) were measured from 1966 to 1986 in a well-defined urban area of Southwestern Finland, the Turku Region. The incidence rates increased markedly, and were, in the mid-80s, among the highest in the world: 2.6 per 100 deliveries, 1.8 per 100 diagnosed pregnancies and 153 per 100,000 fertile-aged women. The regional incidence rate exceeded the national one in the 1970s, whereas in the 1980s the regional rate which has levelled-off during recent years has been equal to, and currently even lower than the national one. This suggests that changes in the incidence of EP in urban area(s) preceed those in the whole country. PMID- 2565258 TI - Trophoblastic disease: 20 years' experience. AB - A series of 155 women with gestational trophoblastic disease (GTD) was evaluated to determine the effect of age on the severity of the disease. The malignant sequelae of hydatidiform moles were of similar frequency at all ages. However, young women, less than 20 years old, had significantly less malignant GTD and significantly less metastatic malignant GTD than did older women. PMID- 2565259 TI - Biochemical changes in human cervical connective tissue during pretreatment with laminaria tents in legal first trimester abortions. AB - In 14 legal first trimester abortions, vacuum aspiration was preceded by dilatation of the cervical canal by laminaria tent for 16-20 h. Biopsy specimens obtained from the cervix before insertion and before removal of the laminaria tent were analyzed for biochemical changes in cervical connective tissue. Pretreatment with laminaria tents resulted in a decrease of the total collagen concentration and of the collagenolytic activity, though the differences were without statistical significance. Both these findings and those concerning the subfractions of the hydroxyproline concentration suggest that pretreatment with laminaria induces a certain degree of softening of the cervix, similar to that reported after intracervical application of PGE2 gel. PMID- 2565260 TI - Sexually transmitted disease among women attending a family planning clinic in Zaria, Nigeria. AB - Three types of sexually transmitted infections were investigated among 150 family planning clients using different methods of contraception and 50 controls. For oral contraception, intrauterine contraceptive device, medroxyprogesterone acetate (depo-provera) and controls, the incidence of trichomonas was 0%, 6%, 8% and 4%; for gonorrhoea it was 0%, 8%, 2% and 6%; for syphilis it was 4%, 34%, 16% and 12%. Only the difference in prevalence of syphilis among intrauterine contraceptive users and control was statistically significant (P less than 0.3). PMID- 2565261 TI - Sheep-acquired severe Chlamydia psittaci infection in pregnancy. AB - There have been five confirmed cases of severe Chlamydia psittaci infection during pregnancy, three having been treated in Edinburgh, Scotland. The most recent case is presented and previous experience is reviewed. The illness usually causes thrombocytopenia with disseminated intravascular coagulation, renal failure and hepatic dysfunction during the late second and early third trimester. The outcome for the fetus is usually fatal and the infection only resolves after delivery or abortion. The main hope is for education to prevent infection occurring in susceptible populations. PMID- 2565262 TI - Sarcoidosis of the female genital tract: a case presentation and survey of the world literature. AB - Sarcoidosis of the female genital tract is a rare clinical entity with only 20 cases reported in the world literature to date. An additional case is presented with a review of the previously reported cases. The diagnostic and histologic aspects of the disease are also discussed. The presence of granulomatous diseases in the female genital tract should initiate a thorough investigation for potential etiologies by both the pathologist and clinician. Etiologies of granuloma fraction must include coccidiomycosis, foreign body reactions, lymphogranuloma inguinale, and tuberculosis. Bacteriologic proof is essential to differentiate these from sarcoidosis. PMID- 2565263 TI - Leiomyoma of the female urethra--a clinical curiosity. AB - A case of leiomyoma of the female urethra is reported. The patient, a perimenopausal, multiparous woman presented with a gradually enlarging mass per vagina for the past 6 months. A provisional diagnosis of carcinoma of skene's duct or vaginal leiomyoma was entertained and an excisional biopsy performed. To our surprise, histopathological examination revealed an urethral cellular leiomyoma. PMID- 2565264 TI - Clinical pharmacology of antipsychotic drugs. AB - Antipsychotic drugs are widely used to treat abnormal behaviour particularly that related to the functional psychoses such as schizophrenia. This review discusses the pharmacokinetics and pharmacology of antipsychotic drugs like chlorpromazine. Clinical use comprises the induction of tranquillization in disturbed psychiatric patients, the treatment of acute and chronic schizophrenic symptoms, and the postponement of relapse in such patients. Unwanted effects are multifarious, involving many systems of the body. Extrapyramidal signs and symptoms are particularly noticeable, and the chronic type, tardive dyskinesia, is a major problem. PMID- 2565265 TI - Pharmacological modulation of cutaneous reactivity to histamine: a double-blind acute comparative study between cetirizine, terfenadine and astemizole. AB - In a double-blind study performed in 81 healthy volunteers, 10 mg cetirizine and 60 mg terfenadine given orally in a single administration significantly inhibited skin reactivity to histamine. Astemizole (10 mg) was completely ineffective. The inhibitory effect of cetirizine was potent and regular whereas 6/28 (21%) volunteers did not respond to terfenadine. The difference observed between cetirizine and terfenadine might be due to differences in the metabolism of the two drugs after administration: terfenadine is rapidly and extensively metabolized whereas cetirizine is directly active without the need for biotransformation and, indeed is poorly metabolized. PMID- 2565266 TI - Ranitidine plus bromazepam in the treatment of duodenal ulcer: effect on gastric acid secretion. AB - The effect of adding bromazepam to treatment of duodenal ulcer with ranitidine was investigated in 30 out-patients. Under double-blind conditions one group of 15 patients received, for 14 days, 300 mg ranitidine and 6 mg bromazepam in the evening as a single dose. The other group received the same dose of ranitidine together with placebo, also for 14 days. In addition to measurements of gastric acid secretion after fasting and following histamine provocation, psychological ratings (Hamilton Anxiety Rating Scale and Zung Self-Assessment Scale for Anxiety) were carried out. Basal acid secretion was the same in both groups. During treatment, maximal acid output was significantly lower in the group of patients taking bromazepam than in those taking placebo. Similarly, significant differences were seen in favour of bromazepam in the psychological tests. It is concluded that these results confirm that bromazepam exerts a significant influence on acidity, over and above the effect of ranitidine. This effect, in all probability, is modulated by the emotional environment of individuals. PMID- 2565267 TI - Oral bioavailability of nizatidine and ranitidine concurrently administered with antacid. AB - This four-way crossover open-label study on eight healthy subjects was designed to investigate the effect on bioavailability of orally administered nizatidine given concurrently with an antacid (magnesium hydroxide-aluminium hydroxide mixture). Ranitidine was used as a comparison. Subjects were hospitalized overnight for each drug application which was given as a single dose [nizatidine or ranitidine (300 mg) or antacid (20 ml) of a neutralizing capacity of 50 mmol hydrochloric acid]. Plasma nizatidine or ranitidine concentrations were measured by high performance liquid chromatography. No statistically significant difference occurred in the kinetic profile of nizatidine after antacid administration although it took longer to reach maximum concentration in plasma. The area under the concentration-time curve was also reduced (by less than 10%) and there was a longer lag between administration and absorption. Bioequivalence (Westlake test) for ranitidine in the presence or absence of antacid was confirmed (difference less than 12%). For nizatidine, with or without antacid, the value of the Westlake test was just above the limit for bioequivalence (21.4%), whereas the Student's t-test for related means (one-tailed distribution) gave P = 0.045. There was no clinically relevant nizatidine-antacid interaction at the doses used in this study. PMID- 2565268 TI - Genetic analysis of the homeotic gene complex (HOM-C) in the beetle Tribolium castaneum. AB - Our laboratories have undertaken both genetic and molecular studies of the homeotic gene complex (HOM-C) of the beetle Tribolium castaneum, and this paper discusses results from our genetic analyses. We describe here the adult phenotypes and complementation behavior of over 50 new mutations. Many of these homeotic phenotypes resemble those of Drosophila melanogaster, but few precisely parallel the segmental transformations seen in this fly. Analysis of putative loss-of-function mutations affecting the head and thorax suggests that the maxillopedia and Cephalothorax genes most closely resemble proboscipedia and Sex combs reduced of Drosophila. In the abdomen, putative loss-of-function alleles of Abdominal affect a domain corresponding to those of the combined abdominal-A and Abdominal-B genes of Drosophila. In contrast to the situation in flies, Abdominal loss-of-function variants in Tribolium cause anteriorward transformations in A3 A5a, but posteriorward transformations in A5p-A7. The implications of the differences in developmental strategies evolved in Tribolium vs Drosophila are discussed. PMID- 2565270 TI - [Hepatitis B virus and periarteritis nodosa]. PMID- 2565271 TI - [The liver and periarteritis nodosa]. PMID- 2565272 TI - Opioid-like action of eseroline on micturition reflex in rats. AB - 1. Eseroline (3-10 mg/kg s.c.), the first product of metabolic breakdown of eserine (physostigmine) markedly increased bladder capacity and decreased voiding efficiency of reflex micturition in rats. 2. Eserine had an opposite effect on bladder function in vivo, while morphine (1-3 mg/kg s.c.) reproduced the action of eseroline. The effects of eseroline or morphine were prevented by naloxone which per se reduced bladder capacity. 3. In vitro both eserine and eseroline potentiated the cholinergic component of the twitch response to field stimulation of the rat bladder and this effect was atropine-sensitive while morphine had no effect. 4. These findings provide further support to the notion that eseroline possesses both eserine- and opioid-like actions, but this latter predominates in determining its actions on bladder function in vivo. PMID- 2565269 TI - The genetic predisposition to fibrocalculous pancreatic diabetes. AB - Fibrocalculous pancreatic diabetes (previously known as tropical pancreatic diabetes) is a rare cause of diabetes confined to countries within the tropical belt. The aetiology of fibrocalculous pancreatic diabetes is thought to be environmental although the agent(s) is unknown. We have investigated a possible genetic basis of this disease by looking for restriction fragment length polymorphisms of genes implicated in the aetiology of diabetes mellitus. Seventy six Dravidian patients with fibrocalculous pancreatic diabetes were studied, and the restriction fragment length polymorphisms obtained compared to racially matched control subjects (n = 94), patients with Type 2 (non-insulin-dependent) diabetes (n = 87) and Type 1 (insulin-dependent) diabetes (n = 58). No association of fibrocalculous pancreatic diabetes was found with restriction fragment length polymorphisms of the insulin receptor gene. Although no association of fibrocalculous pancreatic diabetes was found with polymorphism of the HLA DR alpha/DQ alpha/DX alpha genes, an association was found with the Taq 1 restriction fragment length polymorphisms of the DQ beta gene (DQ beta T2/T6 present in 39% of patients with fibrocalculous pancreatic diabetes compared to 19% in control subjects; p = 0.01; corrected p value = 0.04) which is similar to that found in Type 1 but not Type 2 diabetes. An association of fibrocalculous pancreatic diabetes was also found with the hypervariable region in the 5-prime flanking region of the insulin gene; 40% of patients possessed the class 3 allele compared to 9.5% of control subjects p = 0.0001; corrected p value = 0.0008).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565273 TI - Involvement of threshold level in the contractile responses for some alpha 1 andrenoceptor agonists in the rabbit iris dilators. AB - 1. The relationship between receptor occupancies and contractile responses for some alpha 1-adrenoceptor agonists were investigated in rabbit iris dilator smooth muscles. 2. Noradrenaline acted as a full agonists, while oxymetazoline and xylometazoline behaved as partial agonists with moderately higher intrinsic activity, and tizanidine and clonidine were partial agonists with lower intrinsic activity. 3. The pD2-values of oxymetazoline and xylometazoline were practically equal to the corresponding pKB-values, the negative log of dissociation constant, estimated by the partial irreversible blockade of alpha 1-adrenoceptors with phenoxybenzamine. However, the pD2-values of tizanidine and clonidine were significantly lower than the corresponding pKB-values. 4. The threshold phenomena lay between the receptor occupations and tissue responses, therefore, the pKB values of partial agonists with lower intrinsic activity were different from their pD2-values. 5. These results suggest that the threshold phenomena in the tissue used may be an important factor in determining the agonist sensitivity. PMID- 2565274 TI - Xylamine enhances pineal gland N-acetyltransferase activity in vitro. AB - 1. The action of N-2'-chloroethyl-N-ethyl-2-methyl benzylamine (xylamine) on rat pineal gland sympathetic innervation was examined. 2. This alkylating agent caused a concentration-dependent increase in pineal gland N-acetyltransferase (NAT) activity in neurologically intact pineal glands that was suppressed in glands previously subjected to bilateral superior cervical ganglionectomy. 3. Xylamine-induced elevations in NAT activity were attenuated by beta-noradrenergic antagonist drugs but not by alpha-noradrenergic antagonist drugs. 4. Since pineal gland uptake of radiolabelled norepinephrine (NE) was impaired by xylamine, the drug may increase pineal gland NAT activity by inhibiting NE reuptake into the presynaptic nerve terminal, thereby increasing the amount of the neurotransmitter available to stimulate pinealocyte beta-noradrenoceptors. PMID- 2565275 TI - Endothelium modulates the effects of alpha-adrenoceptor agonists in vascular smooth muscle. AB - 1. The contraction induced by clonidine and guanfacine but not phenylephrine was enhanced in endothelium-denuded and methylene blue pretreated rabbit aortic, pulmonary artery rings and isolated perfused whole superior mesenteric, carotid and femoral arteries from the same species. 2. The responses to acetylcholine in guanfacine preconstricted superior mesenteric and carotid arteries were also augmented when compared with phenylephrine and clonidine preconstricted segments. No difference was observed to the relaxing activity of acetylcholine in the aortic, pulmonary artery rings and whole perfused femoral artery contracted by phenylephrine, clonidine and guanfacine. 3. Acetylcholine also produced a biphasic effects in the pulmonary artery rings precontracted with the used alpha adrenoceptor agonists. The contractile effect was observed with the concentration up to 10(-6) M of acetylcholine and was higher for guanfacine than phenylephrine and clonidine precontracted rings. 4. These results were taken as an evidence for the specificity of alpha 2-adrenoceptor agonists on the production and release of EDRF from vascular endothelium and in this respect guanfacine seems to have higher potency than clonidine. 5. These results also indicate that production or release of EDRF from vascular endothelium may vary depending on the regional vascular bed. PMID- 2565276 TI - In vitro release of pancreatic hormones following 99% pancreatectomy in the chicken. AB - In vitro assessment was made of the hormone-release capability of splenic pancreatic tissue 16 days after adult chickens had 99% of the pancreatic mass surgically removed. The objective of this study was to evaluate if the enlargement of the splenic lobe remnant after 99% pancreatectomy was attended by alterations in the responsivity of hormone release and, if so, were such changes reflective of all pancreatic hormones. After a 24-hr fast, splenic lobe tissue was obtained from young adult chickens on Postoperative Day 16, diced into 18-22 mg cubes, and incubated in vitro in media containing varying amounts of glucose with or without added somatostatin (SRIF). At 15-min intervals, the tissue cubes were transferred to fresh media and samples of each medium measured for insulin, glucagon, and APP. Viability of the tissue after 75 min was tested by tissue response to added 5 mM phenylalanine. The results obtained indicated that while total content of all four hormones (including SRIF) increased with tissue enlargement, the concentration of each decreased significantly except for SRIF, which remained at control levels. Further, the sensitivity of the B-cell in releasing insulin when confronted by a glucose challenge was not altered by previous pancreatectomy, while that of glucagon release from the A-cell was depressed. A-cell responsivity to SRIF does not appear to be adversely affected by previous 99% pancreatectomy. APP release was least affected by SRIF addition to the media, although depression by high glucose occurred. It is concluded that differential alterations occur in chicken pancreatic hormone-releasing cells as a result of 99% pancreatectomy. The efficacy in maintaining low, but still adequate, plasma I/G molar ratios (reported earlier) by the splenic remnant tissue either reflects a remarkable functional readjustment to surgical removal of 99% of the pancreatic mass in chickens or, alternatively, suggests the existence of extrapancreatic sources of insulin and glucagon, but not APP. PMID- 2565277 TI - The segmentation gene Kruppel of Drosophila melanogaster has homeotic properties. AB - In Drosophila hindgut, Malpighian tubules and posterior midgut develop from the most posterior region of the blastoderm. One of the genes that influences the differentiation of the Malpighian tubules is Kruppel (Kr), a segmentation gene of the gap class. Kr homozygous embryos lack thoracic and abdominal segments and, depending on the allele, develop nearly normal Malpighian tubules or do not differentiate them at all. In the wild type, injection of horseradish peroxidase (HRP) into cells of the early gastrula at various posterior positions results in labeling of hindgut (93%), Malpighian tubules (46%), and posterior midgut (20%). Malpighian tubules were labeled only in combination with hindgut. In Kr1 homozygous embryos that lack Malpighian tubules, the label was restricted to hindgut (84%) and posterior midgut (24%). Because we could not find significant cell death in the posterior region of Kr1 embryos, we counted the cell nuclei in the hindguts of wild-type and mutant embryos. The results show that the hindgut in Kr1 embryos contains those cells that would differentiate into Malpighian tubules in wild type. Therefore, we conclude that the Kruppel gene exhibits a homeotic function in addition to its role as a segmentation gene and is involved in separating hindgut and Malpighian tubule cells and in the elongation process as well. PMID- 2565278 TI - A new family of mouse homeo box-containing genes: molecular structure, chromosomal location, and developmental expression of Hox-7.1. AB - Two families of homeo box-containing genes have been identified in mammals to date, the Antennapedia- and engrailed-like homeo boxes, based on the sequence similarity to those from Drosophila. Here, we report the isolation of a homeo box containing gene that belongs to a new family of which there are at least three related genes in the mouse genome. The homeo box of this new gene shows remarkable similarity to the Drosophila Msh homeo box that we designate as the prototype for this family. The gene maps to the proximal end of mouse chromosome 5 and does not cosegregate with any known homeo box-containing gene. We designate this locus Hox-7.1. In situ hybridizations to mouse embryos at different stages show a unique pattern of expression, as compared to other homeo box-containing genes described thus far. Hox-7.1 transcripts are detected in 9.5-day-old embryos in the neural crest, developing limb bud, and visceral arches. Later, this gene is expressed in regions of the face that are derived from neural crest and in the interdigital mesenchymal tissues in both the fore- and hindlimbs. PMID- 2565280 TI - [Syrup of ipecac as an emetic]. PMID- 2565279 TI - Immunoglobulin secretion by isolated intestinal lymphocytes: spontaneous production and T-cell regulation in normal small intestine and in patients with coeliac disease. AB - The in vitro secretion of immunoglobulins by small intestinal lymphocytes isolated from 47 patients with normal histology and 23 patients with treated and untreated coeliac disease was examined using an enzyme linked immunosorbent assay. In control patients, duodenal lymphocytes spontaneously secreted higher levels of IgM than jejunal lymphocytes (p less than 0.05). Significantly higher levels of both IgA (p less than 0.05) and IgM (p less than 0.001) were secreted by jejunal lymphocytes of 10 patients with untreated coeliac disease than cells isolated from normal jejunal tissue. IgM and IgA secretion by duodenal lymphocytes isolated from control patients was increased in a dose dependent manner by coculture with autologous peripheral blood T lymphocytes. This effect was not observed with jejunal lymphocytes of control or treated coeliac patients. Peripheral T-cells of untreated coeliac patients, however, showed significant helper effects (p less than 0.05) for IgM and IgA secretion by autologous jejunal lymphocytes. The results suggest that jejunal lymphocytes of patients with untreated coeliac disease show major differences in their capacity to synthesise and secrete immunoglobulins in vitro and the enhanced secretion might result from changes in T-cell immunoregulatory function. PMID- 2565281 TI - [Benzodiazepines--good hypnotics? Interview with R. Steinberg, Landeck Pfalz clinic: suppression of deep sleep-dependence in long-term use]. PMID- 2565282 TI - [Kaposi sarcoma. T4 lymphocytes secrete growth factor]. PMID- 2565283 TI - [Epileptic psychoses and their drug treatment]. AB - This paper is concerned with classification, clinical-electroencephalographic correlation, principles of treatment, and pharmaceutic therapy of epileptic psychoses. Based on the system of the physically founded reversible psychoses, classification of epileptic psychoses is developed, which is easy to apply for clinical and research purposes. Its principles are the criteria of disturbance of consciousness and of connexion to epileptic seizures. Epileptic psychoses without disturbance of consciousness frequently go along with a forced normalization of epileptic EEG-changes. This clinical-electroencephalographic correlation is documented by the cases of a depressive-paranoid and a cenesthetic alternative psychosis. Epileptic psychoses connected to seizures, going along with disturbances of consciousness, however, show, without any exception, a pathological changed EEG. Also in the cases of the often iatrogenically produced epileptic psychoses with disturbances of consciousness yet not connected to seizures, the EEG-results are of decisive diagnostic importance. Each of these three clinical-electroencephalographically defined groups of psychoses calls for concentration on particular pathogenetical aspects concerning a specific pharmaceutic therapy. The respective principles of treatment are developed in subtly differentiated ways and they are provided with suggestion as to medicamental treatment. Schizophrenia-like epileptic psychoses are a model for idiopathic schizophrenias and so important perspective opens up for research. PMID- 2565284 TI - Effects of transglutaminase substrates and inhibitors on the motility of demembranated reactivated spermatozoa. AB - The effects of transglutaminase (TGase) substrates putrescine, dansylcadaverine, spermine, etc., and the TGase inhibitor cystamine were tested on the motility of demembranated mammalian spermatozoa. These products blocked within a few seconds the motility of demembranated reactivated spermatozoa at concentrations ranging from 0.25 to 5 mM. These minimal inhibitory concentrations could be decreased 5 150-fold when TGase substrates and inhibitor were incubated with demembranated spermatozoa for 15 min prior to the addition of Mg.ATP. The inhibition was reversed by higher concentrations of Mg.ATP but none of these TGase substrates or inhibitor could inhibit bull sperm dynein ATPase. TGase activities, as measured by the incorporation of 3H-putrescine into TCA-precipitable proteins, were present in both sperm Triton-soluble and -insoluble fractions. On the other hand, amine acceptor protein substrates for the TGase-catalyzed reaction were present only in the insoluble fraction. The Triton-soluble TGase was similar to the known "tissue" TGases; the Triton-insoluble TGase activity was calcium independent. The same TGase substrates and inhibitor that blocked the motility of reactivated spermatozoa also blocked TGase activities. Linear relationships were observed between the concentrations of these substances required to block sperm motility and those to block TGase activities. These data suggest the involvement of a TGase activity in sperm motility. PMID- 2565286 TI - What is the place of laser lithotripsy in the treatment of common bile duct stones? PMID- 2565285 TI - Hepatitis B surface antigen binds to human serum albumin cross-linked by transglutaminase. AB - It has been postulated that polymerized human serum albumin may play a role in the infection of hepatocytes by hepatitis B virus, because both the envelope of hepatitis B virus (HBsAg) and hepatocytes exhibit binding activity for human serum albumin after cross-linking by glutaraldehyde. Since glutaraldehyde dependent cross-linking of albumin molecules is not likely to occur in vivo, we considered the possibility that albumin may be polymerized by the action of transglutaminase enzymes present in plasma as activated factor XIII or released into plasma from tissues. Guinea pig liver transglutaminase covalently cross linked human serum albumin molecules into dimers, trimers and polymers up to hexamers as shown by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. HBsAg particles bound transglutaminase-cross-linked as well as glutaraldehyde-cross-linked human serum albumin as demonstrated by radioimmunoassay and immunoelectron microscopy. The binding was blocked by preincubation of HBsAg with transglutaminase- or glutaraldehyde-cross-linked human serum albumin, anti-HBs or monoclonal anti-pre-S2, but not by polymerized bovine or rat serum albumin or by monomeric human serum albumin. These data indicate that HBsAg particles contain specific binding sites for transglutaminase cross-linked human serum albumin, but it remains to be determined whether the albumin polymers play a role in the attachment of hepatitis B virus to hepatocytes. PMID- 2565287 TI - Glutamate and kainate increase intracellular sodium activity in leech neuropile glial cells. AB - Na+-selective, double-barrelled microelectrodes were used to measure intracellular Na+ activity (aiNa) and membrane potential (Em) in neuropile glial cells of isolated segmental ganglia in the leech Hirudo medicinalis. Bath application of glutamate (10(-3) M) resulted in membrane depolarizations of about 5 mV and a concomitant increase of aiNa by between 2 and 10 mM. Kainate (10(-4) M) elicited depolarizations of up to 40 mV amplitude followed by a prominent after hyperpolarization. During kainate, aiNa increased by 7 to 25 mM. In contrast to glutamate, an initial decrease of aiNa was detected during the action of kainate. N-methyl-D-aspartate (NMDA, 10(-5)-10(-3) M) had no effect of Em and aiNa. The results indicate that leech glial cells have a kainate-preferring non NMDA glutamate receptor. PMID- 2565288 TI - Activation of excitatory amino acid receptors reduces thymidine incorporation and cell proliferation rate in primary cultures of astrocytes. AB - Addition of quisqualate (a heterocyclic analogue of glutamate) reduced [methyl 3H]thymidine incorporation and cell proliferation in primary cultures of rat cortical astrocytes. The inhibitory action of quisqualate was mimicked by glutamate and ibotenate, whereas kainate, N-methyl-D-aspartate (NMDA) and alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) were inactive. These results suggest that activation of a specific class of excitatory amino acid receptors contributes to the regulation of growth and proliferation of glial cells in primary culture. PMID- 2565289 TI - Variable expression of a yeast-phase-specific gene in Histoplasma capsulatum strains differing in thermotolerance and virulence. AB - By means of differential hybridization techniques, several yeast-phase-specific DNA sequences were identified in the dimorphic pathogenic fungus Histoplasma capsulatum. A 1.85-kilobase (kb) HindIII fragment from one genomic clone, yps-3, hybridized to at least three distinct yeast poly(A)+ RNAs of 1.3, 1.05, and 0.95 kb from the virulent strain G217B. These mRNAs were not detected in mycelia. When mycelia from G217B were induced to become yeast by transfer from 25 to 37 degrees C, a process requiring approximately 9 days, expression of yps-3 was detected within 24 h, although not in the initial 2 h following the temperature shift. In contrast, a low-virulence strain (Downs) which completes the transition in approximately 2 weeks failed to express the yps-3 gene during phase transitions. A third isolate, G186B, intermediate in its virulence properties and in the time required for the transition (11 days), expressed a single 1.25-kb mRNA but only at low levels in the yeast phase and only after 3 days following the 25-to-37 degrees C temperature shift. Although yps-3 expression does not appear to be essential for the transformation to the yeast phase, it may facilitate the early adaptive processes which permit the mycelium-to-yeast transition and survival of the yeast phase of H. capsulatum at elevated host temperatures. The phase specific yps-3 nuclear gene is carried on highly polymorphic restriction fragments in all three strains, suggesting that this probe may provide a sensitive diagnostic tool for the classification of H. capsulatum isolates. PMID- 2565290 TI - Use of mitochondrial and ribosomal DNA polymorphisms to classify clinical and soil isolates of Histoplasma capsulatum. AB - We have developed an improved scheme for the classification of environmental and clinical isolates of Histoplasma capsulatum that is based on analysis of mitochondrial DNA (mtDNA) and ribosomal DNA (rDNA). Strains were initially divided into mtDNA groups according to restriction digests of whole-cell DNA and Southern hybridization with cloned mtDNA probes. Strains within a mtDNA class could be further grouped by polymorphisms in rDNA. The majority of soil and clinical isolates from the United States had identical mtDNA patterns; however, rDNA polymorphisms were common in both types of isolates. The combination of mtDNA and rDNA typing described in this report will be useful in resolving questions concerning the epidemiology of H. capsulatum infections. PMID- 2565291 TI - Cloning and expression of an adhesin (AIDA-I) involved in diffuse adherence of enteropathogenic Escherichia coli. AB - The adherence of enteropathogenic Escherichia coli (EPEC) to the small bowel mucosa is an important step in the pathogenesis of diarrheal diseases. It has been shown that many EPEC strains adhere to HEp-2 and especially HeLa cells in characteristic patterns termed localized adherence (LA) and diffuse adherence (DA). A plasmid-derived DNA fragment encoding a factor specific for LA hybridized only to EPEC strains expressing LA, which demonstrated that LA and DA are mediated by two genetically distinct adhesins. EPEC strain 2787 (O127:H27), isolated from a case of infantile diarrhea, exhibited three major properties: (i) it showed DA to HeLa cells, (ii) it carried two large (ca. 100-kilobase [kb]) plasmids and one small plasmid of about 3 kb, and (iii) no fimbriae could be detected by electron microscopy in organisms grown on agar plates or in liquid cultures. Whole isolated plasmid DNA was partially digested with EcoRI and cloned into the vector pBR322. One recombinant clone (pIB6) was found to exhibit the same DA pattern on HeLa cells as did the parent strain. This clone contained an 11-kb DNA fragment derived from the largest of the three plasmids, as shown by Southern hybridization. By deletion analysis, a 6.0-kb DNA fragment was shown to be sufficient for expression of the DA phenotype. This insert encoded the production of a 100,000-dalton protein mediating adhesion to HeLa cells. PMID- 2565292 TI - Genetic relationships among Mycobacterium leprae, Mycobacterium tuberculosis, and candidate leprosy vaccine strains determined by DNA hybridization: identification of an M. leprae-specific repetitive sequence. AB - Comparative DNA hybridization studies of genomic DNA indicated that, while different isolates of armadillo-derived Mycobacterium leprae have a high degree of homology, binding of M. leprae genomic DNA to DNA of other species of mycobacteria or to corynebacteria was low, establishing that M. leprae is only remotely genetically related to any of the species examined. Several candidate leprosy vaccine mycobacterial strains were similarly found to have little genetic similarity to M. leprae. In contrast, the DNAs of the slow-growing mycobacteria M. tuberculosis, M. africanum, M. bovis, and M. microti were found to be very closely related. In the course of these studies, an M. leprae-specific repetitive sequence, greater than 15-fold per genome equivalent, was identified that might be useful for diagnostic and epidemiological studies. PMID- 2565293 TI - Heterologous protection against invasive Escherichia coli K1 disease in newborn rats by maternal immunization with purified mannose-sensitive pili. AB - Heterologous protection against Escherichia coli K1 bacteremia with antibody to purified mannose-sensitive (MS) pili was demonstrated in a neonatal rat model. The serological relatedness of purified MS pili from 17 E. coli K1 clinical isolates was examined by an enzyme-linked immunosorbent assay. Five pilus serogroups were identified, with the pili in each group showing 50% or greater cross-reactivity with the typing serum of the group. The MS pili from 12 of 17 (70%) strains belonged to just two serogroups. Pregnant Sprague-Dawley rats (dams) were immunized with purified pili, and their newborns (pups) were challenged with heterologous E. coli. Bacteremia was significantly reduced when the pili used for immunization were from the same serogroup as the pili expressed by the challenge bacteria. Thus, immunization with C94 pili and challenge with E03 (71% cross-reactivity) or E04 (50% cross-reactivity) resulted in bacteremia rates of 12 of 17 (17%) versus 51 of 79 (65%) in controls and 0 of 75 (0%) versus 28 of 70 (40%) in controls, respectively (P less than 0.001 for each comparison). With lower cross-reactivity, less protection was observed (P less than 0.05 for 22 to 37% pilus serological relatedness). No protection was seen in pups suckled by dams immunized with MS pili having only 5% serological relatedness to the pili on the challenge strain. PMID- 2565295 TI - Role of interleukin 4 and gamma interferon in the regulation of human IgE synthesis: possible alterations in atopic patients. AB - The IgE helper function of human T cell clones or their phytohemagglutinin induced supernatants was positively correlated with their ability to produce or their content in interleukin 4 (IL-4), whereas it was inversely correlated with production of or content in gamma interferon. The addition to B cell cultures of anti-IL-4 antibody abolished not only the IgE synthesis induced by recombinant human IL-4, but also that induced by IL-4-producing T cell clones or their phytohemagglutinin-induced supernatants. A clonal analysis in nonatopic donors and patients with common atopy showed that atopics possess in their peripheral blood significantly higher numbers of T cells able to secrete IL-4 and to provide helper function for IgE. PMID- 2565294 TI - Distribution of the P-associated-pilus (pap) region among Escherichia coli from natural sources: evidence for horizontal gene transfer. AB - Variation in chromosomal DNA in Escherichia coli was studied with probes specific for the P-associated-pilus (pap) region. The presence of DNA homologous to pap was determined by dot blots. Variation in the number of copies of pap and in the organization of internal and flanking sequences was determined by Southern blot hybridization. The 229 strains studied were also classified by O:K:H serotyping and multilocus enzyme electrophoresis. There was considerable heterogeneity in the presence of pap and distribution of pap-homologous DNA in these E. coli strains from natural sources. In general, there was less variation in pap among strains of the same specific O:K:H serotype and enzyme electrophoretic type than among random isolates. There were, however, E. coli strains identified as members of the same clone by O:K:H serotyping and enzyme electrophoresis that were pap positive and pap negative or had different Southern blot patterns for the pap probes (pap type). There were also isolates of the same pap type that differed in two of three O:K:H serotype antigens and the majority of enzymes that determined their enzyme electrophoretic type. These latter two observations were interpreted as evidence for the horizontal (infectious) transfer of the pap-homologous sequences among clones of E. coli. PMID- 2565296 TI - Assessment of beta-blocking activity of low-dose bupranolol. AB - In the present study an investigation was made on the pharmacodynamic effect of the beta-blocking agent bupranolol in the low-dose range. Bupranolol is usually given in doses of 100 mg twice daily in the treatment of hypertension, however the dose range between 20 and 100 mg was studied using graded isoproterenol injections in healthy volunteers. A significant beta-1-blocking activity was observed for the 20 mg dose already. This effect was reduced after a treatment of 10 days. The effect increased with the higher doses, there might be a linear correlation between the logarithm of the dose and the reduction of the tachycardia after the isoproterenol injections in the low-dose range. It was concluded that using the safe and sensitive isoproterenol injection method, the clinical effect of very low doses of bupranolol may be demonstrated. The low dose might be useful to reduce the reflex tachycardia seen in the treatment of hypertension with vasodilating drugs. PMID- 2565297 TI - Effect of Biostim (RU 41.740) on natural killer cell generation from bone marrow precursors. AB - We have evaluated the possible effect of RU 41.740 (Biostim), a mixture of two glycoproteins extracted from K. pneumoniae, on the in vitro interleukin-2 (IL-2) induced generation of NK cells from bone marrow (BM) precursors and on the in vivo reconstitution of splenic NK activity in lethally irradiated (9 Gy) and BM reconstituted mice. Our results show that RU 41.740 is able to augment the generation of NK cells when added (1-0.01 micrograms/ml) to normal or 5 fluorouracil-resistant BM, cultured in the presence of recombinant IL-2. Also, in vivo treatment of lethally irradiated mice, transplanted with syngeneic BM cells, with RU 41.740 (1-0.1 mg/kg i.v.) from day 0 through day 4 after BM graft, resulted in a significant augmentation of NK activity reconstitution. PMID- 2565298 TI - "Babylon-scopy" or the semantic confusion in gynecologic endoscopy. PMID- 2565299 TI - Long-term use of an LH-RH agonist in the management of uterine leiomyomas: a study of 17 cases. AB - Seventeen nonmenopausal women with symptomatic uterine myomas, diagnosed by clinical examination and confirmed by pelvic ultrasonography, were treated with a delayed-release microcapsule preparation of D-Trp-6-LH-RH (Decapeptyl) injected intramuscularly, every 28 days. The microcapsules were designed to release 100 micrograms/day for 30 days. The mean duration of treatment was 4.7 months (range, 1-11). In all patients, the pituitary-ovarian axis was suppressed after 1 month of treatment, and mean serum estradiol levels fell to 17.3 pg/mL (range, 5-80). There were no significant changes in serum LH and FSH levels. Fifteen patients (83%) experienced an improvement of such symptoms as abdominal pain and bleeding. Enlarged uteri decreased in 81% of patients during the treatment, and in 38% of them the decrease in uterine volume was more than 50%. Among the 12 myomas found in 10 women, 2 disappeared and 9 decreased in volume during the treatment; for 7 myomas the decrease was more than 50%. After Decapeptyl, eight patients did not require any additional therapy, four underwent surgery, and the others were treated with progestins. The side effects were mild, consisting mainly of hot flushes. Our findings suggest that Decapeptyl may be useful for the treatment of uterine myomas. PMID- 2565300 TI - Factors affecting the success of artificial insemination by frozen donor semen. AB - The records of 227 couples who received artificial insemination by frozen donor semen in 984 treatment cycles were analysed to find out the factors affecting the pregnancy rates. The cumulative pregnancy rate at 6 months was 46.8% and the pregnancy rate per cycle was 10%. The pregnancy rates were adversely affected by (1) increase in the recipient age above 30 years, (2) irregularity of menstrual cycles, (3) the use of clomiphene citrate for induction of ovulation, and (4) low cervical mucus scores. Among our donor semen samples selected with a set of criteria, the sperm counts and initial and post-thaw motility did not affect the pregnancy rates. PMID- 2565301 TI - Social problems of infertile women in India. AB - Female partners of 200 consecutively seen infertile couples attending our infertility clinic were interviewed on a structured, precoded form for the social problems faced by the childless women. One or more social problems were present in 34% couples, of whom 53% faced abandonment. Sixteen percent of the women had a strained relationship with their husbands. Social problems increased with duration of marriage and infertility, while these decreased with increase in age, education, and income of the husband. Similarly, the problems decreased with women's education and income. Only seven couples had requested AID. PMID- 2565302 TI - Successful term pregnancies after tubal anastomosis utilizing contralateral tubal segments. AB - In performing reversal of tubal sterilization, the microsurgeon sometimes has to face difficult anatomical situations due to extensive tubal damage, particularly after laparoscopic unipolar cauterization. In a recent case report, a technique of utilization of contralateral fallopian tube segments in tubal reanastomosis has been described. An intrauterine pregnancy followed, which resulted in a spontaneous abortion at 8 weeks' gestation. We report here our experience in ten cases utilizing a similar technique. Three patients achieved a term pregnancy. One patient achieved three intrauterine pregnancies that ended in first-trimester abortions. It is felt by the authors that this technique should be kept in mind by the practicing microsurgeon for selected cases. PMID- 2565303 TI - Evaluation of stimulated cycles with multiple follicular development but low oocyte retrieval. AB - Factors that may explain a low oocyte recovery rate by laparoscopy were examined. Two different groups of women were evaluated. Group A (n = 56) had zero or one oocyte recovered, despite the aspiration of three or more follicles. Group B (n = 36), had oocytes recovered from more than 75% of punctured follicles. Group B showed significant increases in total estradiol (E2) values during the follicular phase (P less than .05), E2 peak levels (P less than .001), and number of patients with E2 rise the day after hCG administration (P less than .001) compared with group A. However, the number and size of the follicles as determined by ultrasound during the last two pre-oocyte recovery days were not significantly different. On the day of laparoscopy, there was a significant decrease in the number of follicles in group A versus B (P less than .001). In addition, the number of patients with a decrease in E2 values the day of laparoscopy was significantly higher in group A (P less than .001). Together these factors suggest an increased incidence of ovulation prior to laparoscopy in group A. The extent and severity of adhesions were graded as absent, mild or moderate, and severe. No difference was observed between groups. Moreover, comparing the number of follicles visualized before laparoscopy and the number of follicles punctured, the ovarian accessibility was not different (76.6% vs. 79.5%). This suggests that a decrease in access to the ovaries was not the cause of poor oocyte recovery. It is concluded that differences in recovery rates, then, are best attributed to the endocrine milieu of the cycle. PMID- 2565305 TI - Encouraging rates of fertility after ectopic pregnancy. AB - A follow-up study (4 to 8 years) on fertility of 110 patients seeking pregnancy after ectopic pregnancy (EP) showed normal delivery in 65%, recurrent EP in 20%, and infertility in 15%. Delivery rates were better in patients who were under age 30 years, used an IUD at the time of operation for EP, and underwent conservative operative treatment for acute EP. The subsequent intrauterine pregnancy rates of EP patients who had used an IUD at the time of ectopic nidation were the same (92%) as after removal of IUD in the healthy population. It appears that IUD plays a predisposing role in EP only when in use. The study also shows that modern diagnostic and therapeutic procedures are beneficial for improving subsequent fertility rates after EP. These fertility rates are highly encouraging with respect to the massive increase of EP in the Finnish population. PMID- 2565304 TI - Chlamydial serology in women with tubal infertility. AB - The purpose of the study was to determine whether serologic studies for Chlamydia would be helpful in identifying cases in which a tubal factor is responsible for infertility. One hundred and fourteen infertile women, consecutive patients who came to the infertility clinic at the University of Texas Medical Branch at Galveston, had serological tests for Chlamydia and subsequent tubal evaluations by laparoscopy or laparotomy. Seventy-four patients (65%) had positive titers for Chlamydia, of whom 57 (77%) were found to have tubal obstruction. Forty-four had distal tubal obstruction; three had cornual obstruction; and ten had peritubal adhesions. Of the 40 patients with negative titers for Chlamydia, only 14 (35%) were found to have tubal obstruction. The difference in the incidence of tubal disease between women with positive titers and those with negative titers was statistically significant (P less than .002). A significant correlation (P less than .001) was observed between the prevalence of antibodies and distal tubal obstruction. Of the 74 patients with positive titers, 48 (65%) had no history of prior symptomatic pelvic inflammatory disease. These findings suggest that chlamydial infection, as evidenced by positive antibody titers, is associated with a significantly high incidence of tubal infertility and that in the majority of these patients, the prior infection was subclinical and asymptomatic. PMID- 2565306 TI - Clinicopathologic diagnosis of mycoplasma endometritis. AB - Chronic endometritis and its association with cervical colonization by Ureaplasma urealyticum was investigated in a group of 28 infertile patients treated for infertility problems. Twenty had positive cervical cultures for U. urealyticum and eight had negative cultures. There was focal chronic endometritis in 11 of the endometrial biopsies of the 20 cases with positive mycoplasma cultures. Ten of these biopsies were in the secretory phase, and one was in proliferative phase. No case with a negative culture showed endometritis. PMID- 2565307 TI - Assessment of infertility surgery by a pelvic scoring system. AB - A pelvic scoring and staging system previously reported was used to evaluate 77 major microsurgical procedures during a 3-year period. Pregnancy rates by clinical assessment groups were (a) surgery indicated for tubo-ovarian adhesions (SGY:F) = 50%; (b) surgery indicated with other pelvic pathology, such as adnexal masses or myoma (SGY:F/etc) = 23%; and (c) surgery questionably indicated (SGY:?) = 18%. Using the scoring system, pregnancy rates by stage were 75%, 41%, and 14% for Stages I, II, and III, respectively. The pregnancy rate was higher in the pure tubal factor group as compared with those with additional infertility factors. Pelvic scores and stages correlated well with clinical expectations. Life-table analysis of cumulative pregnancy rates showed that all subsequent pregnancies occurred within the 24 months following surgery. A significant correlation noted between pelvic scores and cumulative pregnancy rates illustrates the system's usefulness in predicting reproductive potential postoperatively. This method can be used to assist the surgeon in establishing a couple's prognosis and to compare results between surgeons or different surgical techniques. PMID- 2565308 TI - Evaluation of the preovulatory rise of follicle stimulating hormone and progesterone in normally ovulating women of reproductive age. AB - Sixteen normally ovulating women of reproductive age were evaluated to determine the extent of the preovulatory FSH and progesterone rise in association with the LH surge. Highly significant differences were demonstrated between the baseline and peak preovulatory levels of both FSH and progesterone (P less than .0005). These significant peak ovulatory differences of FSH and progesterone were associated with a 2.5-fold rise (mean difference of 9.06 mIU/mL) of FSH and a 3.0 fold rise (mean difference of 1.94 ng/mL) of progesterone. These small, yet very significant, preovulatory increments of FSH and progesterone are important in the ovulatory process and the formation of an adequate corpus luteum. PMID- 2565309 TI - Late sequelae following laparoscopic female sterilization. AB - A retrospective study regarding long-term sequelae among 818 randomly selected patients undergoing elective laparoscopic tubal sterilization is presented. These procedures were carried out at the Women's Medical Pavilion in Dobbs Ferry, NY, a free-standing surgical unit, between January 1, 1980 and June 30, 1985. A questionnaire was sent to each patient's last known address and, at the same time, attempts were made to contact them by telephone; 623 patients were reached and assessed as to satisfaction with tubal ligation, any ensuing menstrual changes requiring a D&C, and/or psychological problems. Also determined was the number of patients in the series who failed to continue routine gynecological care. It was found that the incidence of dissatisfaction with the operation was 2.9%, of undesired menstrual changes, 4.8%, and of psychiatric morbidity, 8.2%. These results were also correlated with the data of our medical records. In our series, 16.9% of the women failed to continue routine gynecological care. The necessity of thorough counseling by strict guidelines and the need for follow-up are discussed. PMID- 2565310 TI - Effectiveness of silicone sheeting in preventing the formation of pelvic adhesions. AB - The uterine horns of 35 female rabbits were subjected to injury by cautery. Ten days later the adhesions were scored and lysed, and in 30 animals the lesion on one side was covered with a segment of silicone. The contralateral uterine horn served as an internal control. After 5, 10, or 15 days, the silicone was removed (groups A, B, and C, respectively). Five animals (group D) in which no silicone barrier was introduced served as an external control. The rabbits were killed ten days later, and the pelvic adhesions were scored again. A significant reduction in adhesion formation following lysis was found in groups A, B, and C. The largest difference between the treated and the untreated side was obtained in group A. In each of these groups, the mean adhesions score on the treated side was significantly lower following lysis than in group D. PMID- 2565311 TI - Flow cytometric analysis of human spermatozoa treated with antiserum to human seminal inhibin. AB - Inhibin from human seminal plasma is structurally identical to sperm coating antigen. Using the flow cytometric technique it has been demonstrated that there is a positive correlation between initial motility of sperm and the amount of inhibin coated on the spermatozoal surface. PMID- 2565312 TI - Reproductive outcome after multiple ectopic pregnancies. AB - Seventy-six patients with two or more ectopic pregnancies treated at the Department of Obstetrics and Gynecology, University Central Hospital, Tampere, Finland over a period of 14 years (1972-1985) were retrospectively analyzed. Conservative tubal surgery had originally been performed in 57% of patients with a repeat tubal pregnancy, and in 41% of control patients with a single tubal pregnancy. After two ectopic pregnancies, 53 patients were actively trying to conceive. Of these patients, 25% achieved delivery, 40% had a third ectopic pregnancy, and 35% did not conceive. Ipsilateral tubal pregnancy occurred in 83% after salpingotomy, in 88% after fimbrial evacuation, and in 47% after tubal resection. Conservative surgery was performed in 16 patients with only one tube where an ectopic pregnancy occurred; 25% had a term delivery, 25% had a repeat ectopic pregnancy, and 50% did not conceive. Follow-up of 19 patients after three tubal pregnancies showed that 16% delivered, 26% had a repeat tubal pregnancy, and 58% did not conceive. There was no significant difference between fertility results after salpingectomy and those after conservative surgery. PMID- 2565313 TI - Cervicitis as an antecedent to pelvic infection: a review. PMID- 2565314 TI - Empirical therapy of the male with clomiphene in couples with unexplained infertility. AB - Unexplained infertility may be secondary to a cryptic male or female factor. Although most often empirical therapy of the female partner may be attempted, clomiphene has been claimed, after uncontrolled studies, to improve fertility in men with subnormal spermograms. We chose to determine if clomiphene therapy of the male would improve fertility in couples with unexplained infertility despite normal-appearing semen parameters. One hundred husbands were randomized to treatment with clomiphene citrate, 25 mg daily for 25 days with 5 days' rest each month, if their social security numbers ended in an even number or ascorbic acid, 500 mg daily, if ending in an odd number. All female infertility factors had to be meticulously corrected for at least eight cycles for inclusion in the study, along with a minimum of 1 1/2 years' duration of infertility. Within 8 months, 29 of 50 couples (58%) with clomiphene therapy of the male achieved a pregnancy, but only 8 of 50 (16%) with ascorbic acid treatment of the male. There were no appreciable changes in sperm counts, motility, or morphology after either treatment, nor were there any significant differences in semen parameters in those conceiving versus those who did not. Further, improved fertility could not be accounted for by improvement in the hamster ova penetration test. Possibly, clomiphene improves some quality of the sperm that is defective but not measurable by standard androgenologic methods, or it improves some aspect of the seminal plasma. Perhaps, though, the results might be better explained on a psychogenic basis, i.e., clomiphene is a "better" placebo than ascorbic acid.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565315 TI - Return of fertility in various types of IUD users. AB - The assessment of reversibility of a contraceptive method is of importance to both providers and users. Ninety-one women who discontinued IUD use because of desire of pregnancy were followed up for pregnancy and its outcome. Cumulative pregnancy rates at 3, 6, 12, and 18 months after IUD removal were 61.5%, 87.9%, 92.3%, and 96.7%, respectively. Mean interval from IUD removal to conception was 4.4 months (range, 1-18 months). These findings add further evidence that fertility of women is not impaired by use of IUDs as a spacing method. PMID- 2565316 TI - Psychosocial distress and infertility: a review of controlled research. AB - Three hypotheses have been most often cited on the link between infertility and psychosocial distress: (1) psychosocial problems trigger infertility; (2) infertility triggers psychosocial distress; and (3) there is an interactive causal relationship between infertility and psychosocial distress. The controlled research on these three hypotheses was reviewed. The thirty publications that met inclusion criteria provide convincing evidence that, taken as a whole, patients diagnosed and treated in infertility clinics show significantly higher levels of psychosocial distress than do control groups. As well, in general, female patients score higher on psychosocial distress measures than males. However, the authors conclude that research designs to date have failed to control crucial variables that permit conclusive empirical tests of the three hypotheses. The paper closes with recommendations for future research that would accelerate the evaluation of scientific data available on the subject. PMID- 2565317 TI - Serum CA 125 as a marker for patients with external endometriosis. AB - Serum CA 125 levels were assayed in 75 healthy female controls and 34 patients with external endometriosis prior to treatment, during treatment, and after treatment. The results of the assay showed that the mean pretreatment serum CA 125 level of the patients with external endometriosis (80.7 U/mL) was significantly (P less than .001) higher than that of the controls (17.9 U/mL). This difference was even more pronounced for those patients with progressive (stage III or IV) endometriosis (mean = 114.0 U/mL). Thus, the assay of serum CA 125 may be useful as an auxiliary tool in the diagnosis of progressive endometriosis. Although the high pretreatment level of serum CA 125 decreased after danazol treatment of 400 mg/day to within the normal limits, usually within 4 to 12 weeks, laparoscopic examinations indicated that most of the endometriosis cases were not cured until 12-16 weeks of danazol treatment. PMID- 2565318 TI - Chromatographic separation of immunosuppressive factors from seminal plasma from vasectomized boars. AB - Components of boar seminal plasma inhibiting the proliferation of murine and porcine lymphocytes were separated into four peaks by chromatography on Sephacryl S-200. Repeated chromatography of these peaks demonstrated that the substances responsible for inhibition of the blastic transformation of lymphocytes corresponded to components with high molecular weight that during separation were broken down to components with a lower molecular weight; these lower-molecular weight components also inhibited lymphocyte proliferation. The component with inhibitory activity in peak II (molecular weight 70,000) was manifestly different from the component in peak I (170,000). PMID- 2565319 TI - Effects of clomiphene citrate on cervical mucus: analysis of some influencing factors. AB - A multicenter study was performed to investigate the effects of a standard dose of clomiphene citrate (100 mg/day from day 5 of the cycle for five days) on the quality of cervical mucus. A total of 82 cervical mucus samples from 60 infertile patients (15 with secondary amenorrhea, 16 with delayed ovulation, 15 with anovulatory cycles, 14 with oligomenorrhea) was examined on the day of maximum follicular diameter. As a control we studied 54 cervical mucus samples from 47 patients with documented tubal sterility or infertile partners, who had spontaneous cycles with normal rhythm. Cervical mucus was unfavorable in 59% of the treatment cycles compared with 11% of the control cycles. Clomiphene citrate's antiestrogenic effect on cervical mucus was found to be related to the length of time between the last drug administration and the day of maximum follicular diameter (delta days), since cervical mucus was favorable in 64% of the cycles with a delta days more than 6 and in only 16% of the cycles with a delta days of 6 or less. Clomiphene citrate thus seems able to decrease cervical mucus quality. This could be one of the causes of the discrepancy between the rates of ovulation and pregnancy reported in patients treated with this drug. PMID- 2565320 TI - Results of operative pelviscopy in 202 cases of ectopic pregnancy. AB - This study presents 202 cases of ectopic pregnancy treated by pelviscopy over 9 years, from 1978 to 1987. The tube was conserved in 177 patients, and a salpingectomy was performed in 25 cases. The former group includes 24 cases of tubal abortion. Longitudinal salpingectomy, to extract the products of conception, was performed in the remainder of cases (153), in 14 of which tubal rupture had already occurred. Postoperative bleeding demanded repeated pelviscopy in three cases, and laparotomy in two. Postoperative infection required salpingectomy in one case. One patient underwent laparotomy in another hospital, and in two cases, chorionic villi were found within a hematosalpinx at re pelviscopy. The pregnancy rate in 74 patients desiring pregnancy was 57%. The recurrence rate of ectopic pregnancy in both the ipsi- and contralateral tube was 7%. PMID- 2565321 TI - In vitro effect of methionine-enkephalin and thymosin on murine expression of Thy1-2 antigen. AB - The present study investigates the in vitro effect of methionine-enkephalin (Met Enk) on Thy1-2 antigen density and the interaction between this opioid peptide and a thymic hormone (thymosin fraction V (TFV]. In vitro, it was found that Met Enk at various concentrations (28.10(-11), 28.10(-10), 28.10(-9) and 28.10(-8) M) had an increased positive effect on the Thy1-2 antigen density of Balb/C mouse thymocytes. This action was observed at all concentrations of Met-Enk. The addition of TFV to Met-Enk thymocyte cultures showed an additive effect of these two hormones. The physiological significance of these observations is that they indicate an interaction between immune and endocrine system functions. PMID- 2565322 TI - Short-term somatostatin infusion affects T lymphocyte responsiveness in humans. AB - Peripheral blood lymphocytes (PBL) function was analysed in 16 young men with duodenal ulcers after one-hour intravenous infusion of somatostatin (SMS) at a dose of 250 micrograms/h. Proliferative responses of PBL from SMS-treated patients were significantly diminished compared with pre-treatment values, after stimulation with PHA, PWM or Con A. Spontaneous IL-2R expression was moderately increased after SMS infusion but PHA-induced IL-2R expression was not affected by this drug. Alloantigen and autoantigen stimulation of PBL showed no significant changes in the proliferative response after SMS infusion. NK cell activity was similarly unaffected. These observations establish a link between SMS exposure and possible development of immune dysfunction. PMID- 2565323 TI - Microwave attenuation of ethanol-induced interactions with noradrenergic neurotransmitter systems. AB - Research suggests that microwave (MW) irradiation can attenuate ethanol (EtOH) induced hypothermia in a manner that may depend, in part, on noradrenergic (NE) neurotransmitter systems. To investigate this possible interaction, neonatal rats were injected with the neurotoxin 6-hydroxydopamine (6-OHDA) to lesion central NE neurons. When tested as adults, lesioned, MW irradiated rats did not demonstrate the interaction between MW (2.45 GHz, 45 min, specific absorption rate = 0.3 W/kg) and EtOH-induced hypothermia that was seen among control animals. Additional experiments examined MW interactions with centrally and peripherally acting beta-adrenergic antagonists. Acute low-level MW irradiation attenuated EtOH-induced hypothermia in the rat. Pretreatment with 1.0 mg/kg of the centrally active beta-adrenergic antagonist propranolol significantly attenuated the ethanol-induced hypothermia of sham-irradiated (SH-irradiated) rats. There was no consistent effect of propranolol on MW irradiated animals, regardless of dose. Similarly, the degree of hypothermia demonstrated by SH-irradiated controls was significantly attenuated compared to MW irradiated animals by pretreatment with the peripheral beta-adrenergic antagonist CGP-12177 (doses of 0.1, 1.0 and 10.0 mg/kg). In vivo binding data indicates only the highest dose of CGP-12177 to be centrally active. Taken together, the results confirm NE mediation of EtOH induced hypothermia and suggest that MW energy may in some way mimic the role of beta-adrenergic antagonists. PMID- 2565324 TI - The polymorphism of HLA-DR3 in South African populations. AB - The polymorphism of HLA-DR3 was investigated in families and unrelated individuals of three population groups: South African (SA) Negroes, Cape Coloureds and SA Caucasoids. Serological and restriction fragment length polymorphism (RFLP) analysis indicated that DR3 could be subdivided into DRw17 (previously DR3.1) and DRw18 (previously DR3.2). In contrast, the two-dimensional (2-D) gel electrophoresis patterns could not distinguish between the DRB1 gene products of the HLA-DRw17 and DRw18 cells. Two DRB3 variants, correlating with the T-cell defined specificities Dw24 and Dw25 were identified at the genomic and product level. Of ten haplotypes studied with the newly defined HLA-DRw18 specificity, all had the DRB3 RFLP pattern associated with Dw24. HLA-DRw17 was found in all three population groups tested, although in the SA Negroes HLA-DRw18 was the prevalent DR3 subgroup. This subgroup was also present in the Cape Coloureds but was absent in the SA Caucasoids tested. HLA-DRw18 forms part of the most characteristic SA Negro haplotype, Bw42, DQw4, Dw"RSH," while HLA-DRw17 is part of the classic Caucasoid haplotype, B8, DQw2, Dw3. PMID- 2565325 TI - A murine monoclonal antibody that recognizes an extracellular domain of the human c-erbB-2 protooncogene product. AB - A murine IgM monoclonal antibody, designated SV2-61, was generated against human c-erbB-2 gene-transfected NIH-3T3 (SV11) cells. SV2-61 defined a 185-kDa molecule present on the surface of SV11 cells, another line of c-erbB-2 gene-transfected NIH-3T3 (A4-15) cells, and MKN-7 human gastric cancer cell line carrying an amplified human c-erbB-2 gene. The SV2-61-defined antigen was found to show protein kinase activity in vitro. The SV2-61 was reactive with human c-erbB-2 gene-transfected NIH-3T3 cell lines but not with transfectants carrying c-erbB-2 gene mutants which lack a coding region for the extracellular domain. It was reactive with a portion of human epithelial cell lines but not with native NIH 3T3, TGF-alpha-coding gene-, activated c-raf gene- or Ha-ras gene-transfected NIH 3T3 cells, or non-epithelial human cells. These results indicate that the SV2-61 is an antibody which recognizes an extracellular domain of the c-erbB-2 gene product, 185-kDa protein. PMID- 2565326 TI - Study of the accuracy of physical and biochemical markers in semen to detect infectious dysfunction of the accessory sex glands. AB - Infection of the male accessory sex glands may result in impaired secretory function and alteration of the composition of seminal plasma. Using receiver operating characteristic curves and accuracy tests, the power of several biochemical and physical markers was evaluated for their ability to discriminate between semen of infected and noninfected infertile men. The total output of citric acid had the strongest discriminating power, followed by acid phosphatase and gamma-glutamyltranspeptidase. Measurement of the concentration of fructose was found to be nondiscriminatory. PMID- 2565327 TI - Pharmacological alterations of the activity of afferent fibers innervating hair cells. AB - To determine whether some of the substances that may be present in hair-cell sensory organs could affect neural activity in afferent fibers, we examined 56 compounds for the ability to alter the discharge rate of afferent fibers innervating hair cells in the lateral line organ of Xenopus laevis, the African clawed frog. These compounds included amino acids, glutamyl dipeptides, standard neurotransmitter candidates, and other constituents of tissues and body fluids. Substances found to be excitatory included some neutral amino acids (alanine, serine, threonine, asparagine, glutamine, and proline), ATP, carnosine, histidine, and barium chloride. Compounds that suppressed discharge included the aromatic amino acids (phenylalanine, tryptophan, and tyrosine), serotonin, and gamma-glutamyl dipeptides. GABA and acidic amino acids (glutamate, aspartate, and cysteine sulfinate) produced a brief excitation followed by a suppression of discharge rate. Several of these substances were active at sufficiently low concentrations that their presence in body fluids may affect afferent fiber discharge rate under normal or pathological conditions. PMID- 2565329 TI - Effects of beta 1- vs. beta 1 + beta 2-blockade on exercise endurance and muscle metabolism in humans. AB - The effects of beta-blockade on muscle utilization of glycogen and triglycerides, as well as potassium metabolism, were studied in eight healthy male subjects performing long-duration exercise to exhaustion. Subjects were studied after treatment with either placebo (PLAC), beta 1-selective (atenolol, 100 mg/day, AT), or nonselective beta-blockade (nadolol, 80 mg/day, NAD) each for 1 wk according to a randomized, double-blind, cross-over design. NAD and AT caused identical decreases in exercise heart rates, but endurance (71 +/- 8 min with PLAC) decreased significantly more with NAD (-33 +/- 4%) than with AT (-14 +/- 6%). Muscle glycogen breakdown, taking exercise time into account, was unaffected by treatment. In contrast, muscle triglyceride utilization was completely blocked by NAD whereas it was unchanged with AT as compared to PLAC. Adipose tissue lipolysis was inhibited to a similar extent by the two beta-blockers. Serum potassium increased to higher levels at exhaustion and muscle potassium decreased to lower levels with NAD than with AT or PLAC. These results suggest that decreased utilization of muscle triglycerides combined with lack of an enhanced glycogenolysis to compensate as well as alterations in potassium metabolism contribute to the decreased exercise capacity with nonselective beta-blockade compared with beta 1-selective blockade. PMID- 2565328 TI - Endothelin produces pulmonary vasoconstriction and systemic vasodilation. AB - Endothelin is a newly described polypeptide derived from endothelial cells. The effects of porcine endothelin on the pulmonary vascular bed and systemic vascular bed were investigated in the anesthetized, intact-chest cat under conditions of constant pulmonary blood flow and left atrial pressure. Intralobar bolus injections of porcine endothelin (100-1000 ng) produced a mild vasoconstrictor response in the pulmonary vascular bed. The pulmonary vasoconstrictor response to endothelin was not altered when pulmonary vasomotor tone was increased by infusion of U46619. In contrast to this mild pulmonary vasoconstrictor response, endothelin decreased systemic arterial pressure. Moreover, injections of porcine endothelin into the right and left atria produced similar reductions in aortic pressure as well as similar increases in cardiac output and decreases in systemic vascular resistance. The systemic vasodilator response to porcine endothelin was not affected by beta 2-adrenoceptor blockade. The present data suggest that endothelin does not undergo significant first-pass pulmonary metabolism. The pulmonary vasoconstrictor response to bolus injections of porcine endothelin is not altered by changes in pulmonary vasomotor tone. In contrast, endothelin markedly dilated the systemic vascular bed independently of activation of beta 2 adrenoceptors. The present study provides the first report of the activity of endothelin on pulmonary and systemic hemodynamics in vivo. Moreover, the potent vasodilator activity of endothelin in the systemic vascular bed and its weak effect on pulmonary vessels suggest that endothelin may be more important in the regulation of peripheral vasomotor tone than the pulmonary vascular bed. PMID- 2565330 TI - Estimation of the mitochondrial redox state in human skeletal muscle during exercise. AB - The mitochondrial redox (NAD+/NADH) state can be used as a reflection of oxygen availability within the mitochondrion. Previous studies using isolated muscle preparations suggest that active muscle is not hypoxic during lactate production, whereas experiments with humans come to the opposite conclusion. Six men exercised for 5 min at 75% maximal O2 consumption (VO2max) and then at 100% VO2max to exhaustion. Ammonia, oxoglutarate (alpha-ketoglutarate), and glutamate, as well as lactate, were measured in biopsies (vastus lateralis) taken at the end of each exercise. The three former metabolites were used to determine the mass action ratio of glutamate dehydrogenase and thus were used as an estimate of the mitochondrial redox state. Muscle lactate increased (P less than 0.05) to 14.5 and 24.5 mmol/kg wet wt after 75 and 100% VO2max, respectively. At both exercise intensities, muscle ammonia rose (P less than 0.05), glutamate fell (P less than 0.05) to only 30-35% of rest levels, and oxoglutarate declined (P less than 0.05). Despite the high levels of muscle lactate accumulation, the estimated mitochondrial redox rate rose 300% (P less than 0.05) in both exercise bouts. This response should increase the activity of key oxidative enzymes and promote increased VO2. Furthermore the data do not support the concept that muscle lactate is formed because of tissue hypoxia. PMID- 2565331 TI - Cyclooxygenase blockers inhibit ethanol-induced pulmonary vasoconstriction in lambs. AB - We investigated the mechanism of ethanol-induced pulmonary vasoconstriction in lambs, by a pharmacological approach. We chronically instrumented 28 lambs to determine whether phentolamine (alpha-block), propranolol (beta-block), promethazine and cimetidine (H1- and H2-block), high-dose indomethacin, or low- and high-dose meclofenamate (cyclooxygenase block) altered the vasoconstriction. Ethanol alone increased pulmonary vascular resistance from 0.14 to 0.49 Torr.ml 1.kg-1.min (U). Only indomethacin (7-8 mg/kg po) and high-dose meclofenamate (7-8 mg/kg iv) abolished the pulmonary vascular response to ethanol infusion. Pulmonary vascular resistance was 0.14 U after ethanol plus indomethacin and was 0.2 U after ethanol plus high-dose meclofenamate (P = NS vs. base line). Low-dose meclofenamate (2 mg/kg) attenuated the vasoconstrictor response. Systemic vascular resistance increased moderately after ethanol and had a similar pattern of inhibition by cyclooxygenase blockade. Cardiac output and heart rate decreased nearly significantly after ethanol (P less than 0.06), a tendency that was also ablated by cyclooxygenase inhibition. Thus the acute cardiocirculatory response to ethanol involves an intact prostaglandin synthase system in lambs. To our knowledge, these data are the first documentation that cyclooxygenase enzyme blockade can eliminate the acute cardiac and vascular effects of ethanol in a whole-animal system. PMID- 2565332 TI - Hypervariable DNA fingerprinting in Escherichia coli: minisatellite probe from bacteriophage M13. AB - Extensive restriction-fragment-length polymorphism was revealed in Escherichia coli strains by using a region of the bacteriophage M13 genome as a DNA hybridization probe. This variation was observed across natural strains, in clinical samples, and to a lesser extent in laboratory strains. The sequence in M13 which revealed this fingerprint pattern was a region of the gene III coat protein, which contains two clusters of a 15-base-pair repeat. Oligonucleotides made to a consensus of these repeats also revealed the fingerprint profile. While this consensus sequence has significant homology to the lambda chi site sequence, an oligonucleotide made of the chi sequence did not reveal polymorphic fingerprint patterns in E. coli. The strain variation revealed by the M13 and M13 derived oligonucleotide probes will be useful for bacterial characterization and should find use in studies of bacterial evolution and population dynamics. The findings raise questions about what these repeated sequences are and why they are so variable. PMID- 2565333 TI - Genetic diversity of avian and mammalian Chlamydia psittaci strains and relation to host origin. AB - Genetic relationships were reported for Chlamydia psittaci derived from psittacine birds, pigeons, turkeys, humans, cats, muskrats, cattle, and sheep and for C. trachomatis, including representative strains of the three biovars, through physical analysis of genomic DNA including DNA fingerprinting with restriction endonuclease SalI, DNA-DNA hybridization in solution with S1 nuclease, and Southern analysis with genomic DNA probes. A total of 26 strains were divided into four groups of C. psittaci and two groups of C. trachomatis, on the basis of DNA fingerprints. The six groups of Chlamydia spp. were related to host origin: two avian groups (Av1 and Av2), one feline and muskrat group (Fe1), one ruminant group (Ru1), one C. trachomatis biovars trachoma and lymphogranuloma group (CtHu), and one C. trachomatis mouse biovar group (CtMo), although an ovine abortion strain belonged to the avian group Av2. DNA-DNA hybridization assay and Southern analysis with genomic DNA probes indicated three DNA homology groups in the genus Chlamydia: an avian-feline group (groups Av1, Av2, and Fe1), a ruminant group (group Ru1), and a C. trachomatis group (groups CtHu and CtMo). Furthermore, the Southern analysis indicated that the homologous sequences (DNA homology of at least 14%) within the avian-feline group were distributed along the whole genome, whereas the homologous sequences (DNA homology of less than 24%) among the three DNA homology groups were localized in distinct regions of the genome DNA. These results suggest that Chlamydia spp. are derived from a common ancestor and have diverged into various groups showing restricted host ranges as a natural characteristic and that the species C. psittaci should be differentiated into groups related to host origin and DNA homology. PMID- 2565334 TI - Differentiation of arcA, arcB, and cpxA mutant phenotypes of Escherichia coli by sex pilus formation and enzyme regulation. AB - In Escherichia coli, mutations in arcA (dye) or arcB anaerobically derepress the synthesis of a multitude of enzymes of aerobic function, and mutations in arcA or cpxA impair F-pilus formation. It is thought that arcA encodes a promoter recognizing protein, whereas arcB and cpxA encode sensor proteins which interact with the arcA product. In this study we found that anaerobic growth of a wild type F' strain decreased the synthesis of both the enzymes and the pilus. Although the two arcA mutants examined were both anaerobically derepressed in the enzymes and impaired in aerobic pilus formation as expected, one mutant hyperproduced the pilus anaerobically. The two arcB mutants examined showed normal pilus formation when grown aerobically. When grown anaerobically they developed more pili than the wild-type strain did when grown aerobically. When a cpxA mutant was examined for synthesis of two aerobic enzymes, normal regulation was found. The available data suggest the following. The arcA product anaerobically represses certain genes of aerobic function and activates certain genes related to F function. It appears that the arcB product senses the redox or energy state; absence of the gene function shifts the arcA product to the nonrepressive form for enzyme synthesis for aerobic pathways. The cpxA product, on the other hand, senses the sexual state; absence of the gene function shifts the arcA product to the inactive form for F-pilus synthesis. PMID- 2565335 TI - The role of paired basic amino acids in mediating proteolytic cleavage of prosomatostatin. Analysis using site-directed mutagenesis. AB - Many small peptide hormones are synthesized as larger precursors in which the mature hormone sequence is flanked by pairs of basic amino acids. These precursors often undergo extensive post-translational modifications; a critical step in this process is proteolytic excision of the hormone at the paired basic residues. To determine the role of paired basic amino acids as recognition signals for cleavage by processing enzymes, we investigated the heterologous expression of prosomatostatin (the pro-somatotropin release inhibiting factor (pro-SRIF). Pro-SRIF is one of the simplest peptide hormone precursors, possessing a single copy of the 14-residue SRIF peptide at its carboxyl terminus preceded by the least common pair of basic amino acids, Arg-Lys. Employing site directed mutagenesis, we altered the paired basic cleavage site to the more common Arg-Arg and Lys-Arg residues. The native and mutated precursors were expressed in rat pituitary GH3 cells and mouse 3T3 cells using a retroviral vector. Alteration of the paired basic residues had no effect on the specificity of proteolytic cleavage as both the native and mutant precursors were processed with 70 to 80% efficiency in GH3 cells. Surprisingly, when the mutant pro-SRIFs were expressed in 3T3 cells, which do not process the native precursor, the Arg Arg and Lys-Arg precursors were processed with 16 and 20% efficiency, respectively. The role of an acidic compartment in mediating pro-SRIF cleavage was also investigated using low concentrations of the lysosomotrophic drug Chloroquine. Twenty-five microM Chlorquine completely inhibited pro-SRIF cleavage and intracellular storage; the unprocessed precursor was secreted into the medium. We conclude that (i) exposure to an acidic compartment is required for pro-SRIF maturation, and (ii) the conformation of the processing site, rather than the composition of the basic amino acids, defines cleavage specificity by prohormone processing enzymes. PMID- 2565336 TI - Characterization of 17-beta-estradiol-dependent and -independent somatostatin receptor subtypes in rat anterior pituitary. AB - Previous studies from this laboratory showed that treatment with 17-beta estradiol (E2) caused an acquisition of inhibitory effect of somatostatin (SRIF) on prolactin release with an increased number of SRIF-binding sites in the rat anterior pituitary. The aim of this study was to characterize the E2-dependent SRIF receptor in comparison with the E2-independent one, which was expressed in ovariectomized rats. The following observations were obtained: 1) both of the E2 dependent and E2-independent SRIF receptors, measured with 125I-Tyr11-SRIF as a radiolabeled ligand, were enriched in the plasma membrane fraction of the cells, displaying a single class of binding site (E2-dependent: Kd, 32 pM, Bmax, 2.3 pmol/mg protein; E2-independent: Kd, 83 pM, Bmax, 0.26 pmol/mg protein). The ligand binding to both receptors was sensitive to monovalent and divalent cations, and GTP. 2) Among the SRIF analogs tested, the relative potencies of SRIF28 and its analog and cyclosomatostatin compared with SRIF were lower in the E2-dependent receptor than in the E2-independent one. 3) A cross-linking study with N-hydroxysuccinimidyl-4-azido-benzoate revealed that the molecular weight of the cross-linked E2-dependent receptor was approximately 94,000, whereas that of the E2-independent one was 82,000, irrespective of the presence of a reducing reagent. The molecular weight of SRIF receptor from normal male or female rat pituitary was similar to the E2-independent type. 4) Both types of the cross linked SRIF receptors were solubilized by sucrose monolaurate, adsorbed to a wheat germ agglutinin-agarose column, and eluted with N-acetyl-glucosamine. 5) SRIF inhibited the forskolin-stimulated adenylate cyclase activity in the pituitary membranes from E2-treated rats, but it did not in the E2-depleted membranes. These results demonstrate that there are at least two subtypes of SRIF receptor in the rat anterior pituitary, one of which is exclusively expressed by the treatment with E2, and that these subtypes are distinct with respect to ligand binding specificity, molecular weight, and coupling to adenylate cyclase inhibition. PMID- 2565338 TI - Expression of a multidrug resistance-adenosine deaminase fusion gene. AB - A novel fusion gene has been created in which the expression of a dominant selectable marker, the human multidrug resistance gene, is directly linked to the expression of human adenosine deaminase cDNA. The chimeric gene was inserted between the long terminal repeats of a Harvey murine sarcoma virus expression vector and used to transfect drug-sensitive human KB carcinoma cells. Transfectants were selected in increasing concentrations of colchicine and found to contain multiple copies of the intact fusion gene, which is stably and efficiently expressed. A membrane-associated 210-kDa human P-glycoprotein adenosine deaminase fusion protein is synthesized which retains function of the multidrug transporter and also exhibits adenosine deaminase activity. The data indicate that the human multidrug resistance gene may be used as a dominant selectable marker to introduce other genes in the form of gene fusions into cultured cells. PMID- 2565337 TI - Heterogeneity at the 5' end of rat acetyl-coenzyme A carboxylase mRNA. Lipogenic conditions enhance synthesis of a unique mRNA in liver. AB - Multiple forms of acetyl-CoA carboxylase mRNA were previously detected in the mammary gland (Lopez-Casillas, F., Luo, X., Kong, I.-S., and Kim, K.-H. (1989) Gene, in press). We have now established that the rat liver also contains heterogeneous acetyl-CoA carboxylase mRNA populations that differ in the 5' untranslated region. In addition, the liver contains a unique form of acetyl-CoA carboxylase mRNA in which the 5'-nontranslated end differs from the species in mammary gland. The 5' end of this unique species was characterized using a procedure for cloning minute amounts of primer extension products (pAU clones). This procedure should also be useful for obtaining full length clones of other mRNAs. The DNA sequence of pAU clones indicates that this liver-specific acetyl CoA carboxylase mRNA has a 315-base long untranslated region. The first 242 nucleotides replace the 5' end of the predominant acetyl-CoA carboxylase mRNA found in the mammary gland (FL56 type). Under lipogenic conditions the unique liver acetyl-CoA carboxylase mRNA increases and is the major species of acetyl CoA carboxylase mRNA. Livers from rats fed a normal diet and the mammary glands of lactating rats do not contain detectable amounts of the pAU type mRNA. On the other hand, the epididymal adipose tissue from these animals contains mainly the pAU type and only minimal amounts of the FL56 type. The multiple forms of acetyl CoA carboxylase mRNA appear to be generated by differential splicing. In addition, transcription appears to be physiologically regulated by the use of tissue-specific acetyl-CoA carboxylase gene promoters. PMID- 2565339 TI - Structure of the human gene for the proliferating cell nuclear antigen. AB - The proliferating cell nuclear antigen (PCNA, cyclin) was originally defined as a nuclear protein whose appearance correlated with the proliferative state of the cell. It is now known to be a co-factor of DNA polymerase delta and to be necessary for DNA synthesis and cell cycle progression. cDNA clones of human PCNA have been isolated and, using one of these cDNA, we have now obtained from a lambda phage library a clone containing the entire human PCNA gene and flanking sequences. The human PCNA gene is a unique copy gene and has 6 exons. It spans, from the cap site to the poly(A) signal 4961 base pairs. We have identified, in the 5'-flanking sequence, a region with promoter activity, a well as other structural elements common to other promoters. An interesting feature of the PCNA gene is the presence of extensive sequence similarities among introns and between introns and exons. PMID- 2565340 TI - Expression of asparagine independence in variants of ICR 2A haploid frog cells. AB - Properties of the change from asparagine dependence (asn-) to independence (asn+) were investigated in the androgenetic haploid frog cell line ICR 2A. Two types of asn+ variants arose spontaneously during culture. Glutamine-dependent asparagine synthetase (AS) activity, found to be deficient in asn- cells, was repressed by asparagine in one type of variant and expressed constitutively in the other. No quantitative differences in AS-specific DNA sequences or changes in ploidy were evident between asn+ and asn- cells. The asn+ frequency in ICR 2A populations, not dramatically influenced by chemical mutagens, was increased 130-fold by exposure to 5-azacytidine. The methylation of CCGG sequences at the 5' end of the AS structural gene was found to be reduced equally in both types of asn+ variant. These results indicate that decreased DNA methylation is essential but not necessarily sufficient for the expression of AS activity in this frog cell system. PMID- 2565341 TI - Regulation of tissue transglutaminase gene expression as a molecular model for retinoid effects on proliferation and differentiation. AB - Retinoids (structural and functional analogs of vitamin A) are potent antiproliferative agents whose mode of action is poorly understood. It has been suggested that the molecular events that underscore their action involve alterations in gene expression, but no gene has yet been shown to be directly regulated by these molecules. Several years ago, we found that retinoic acid caused an accumulation of the enzyme tissue transglutaminase in murine peritoneal macrophages and in human promyelocytic leukemia (HL-60) cells. We now report that this induction is caused by an increase in the mRNA for this enzyme. Retinoic acid is the only mediator of this induction, since its effects do not depend on the presence of serum proteins. The induction of tissue transglutaminase mRNA is not due to an increase in its stability but to an increase in the relative transcription rate of its gene. We present a model to correlate the retinoid induction of tissue transglutaminase with retinoid effects on cellular growth and differentiation. PMID- 2565342 TI - Dipeptidylpeptidase IV and trypsin-like enzymatic degradation of human growth hormone-releasing hormone in plasma. AB - The plasma enzyme responsible for primary proteolytic cleavage of growth hormone releasing hormone (GRH) at the 2-3 amino acid bond was characterized. Native GRH[GRH(1-44)-NH2 and GRH(1-40)-OH], and COOH-terminally shortened fragments [GRH(1-32)-NH2 and GRH(1-29)-NH2] were rapidly cleaved, while GRH(2-32)-NH2 was not degraded at this site. Moreover, degradation to GRH(3-44)-NH2 was unaffected by an aminopeptidase inhibitor, indicating that this metabolite was generated from a single step cleavage by a dipeptidylpeptidase (DPP) rather than sequential aminopeptidase cleavages. Conversion to GRH(3-44)-NH2 was blocked by diprotin A, a DPP type IV (DPP IV) competitive inhibitor. D-Amino acid substitution at either position 1 or 2 also prevented hydrolysis, characteristic of DPP IV. Analysis of endogenous plasma GRH immunoreactivity from a human GRH transgenic pig revealed that the major peak coeluted with GRH(3-44)-NH2. Native GRH exhibited trypsin like degradation at the 11-12 position but cleavage at the 12-13 site occurred only with GRH(1-32)-NH2 and GRH(1-29)-NH2. Formation of these metabolites was independent of prior DPP IV hydrolysis but was greatly reduced by trypsin inhibitors. Evaluation of plasma stability of potential GRH super analogues, designed to resist degradation by these enzymes, confirmed that GRH degradation in plasma occurs primarily by DPP IV, and to a lesser extent by trypsin-like enzyme(s). PMID- 2565343 TI - Circulating prosomatostatin-derived peptides. Differential responses to food ingestion. AB - Prosomatostatin (pro-S) and its bioactive posttranslational products, somatostatin-14 (S-14), somatostatin-13 (S-13), and somatostatin-28 (S-28), were measured in human plasma by the use of immunoglobulins to the NH2-terminus of S 28 conjugated with agarose to separate them and, thereafter, by RIA with an antiserum recognizing the COOH-terminus of pro-S, and by specific RIA for the NH2 terminus of S-14 and pro-S. In healthy men, mean basal levels of pro-S were 4 pg equivalent S-14/ml; S-14/S-13 combined were 9 pg equivalent S-14/ml; and S-28 levels were 16 pg/ml. After a 700-kcal meal, pro-S, S-14, and S-14/S-13 did not change, whereas S-28 levels doubled by 120 min and remained elevated for 240 min. To evaluate the origins of these peptides, their levels were compared in peripheral, portal, gastric, and mesenteric veins of anesthetized patients and in patients with total resection of stomach and pancreas before and after nutrient intake. The stomach and small intestine were sources of both peptides; however, most S-28 originated in the small intestine. These findings suggest that, in contrast to S-14, S-28 is a hormone and may modulate postprandial nutrient absorption and use. PMID- 2565344 TI - Molecular cloning and nucleotide sequence of complementary DNAs encoding human short chain acyl-coenzyme A dehydrogenase and the study of the molecular basis of human short chain acyl-coenzyme A dehydrogenase deficiency. AB - Complementary DNAs encoding the precursor of human placental short chain acyl coenzyme A (CoA) dehydrogenase (SCAD) (EC 1.3.99.2) were cloned and sequenced. The cDNA inserts in these clones were 1,852 bases in length combined, and encoded the entire 412-amino acid precursor SCAD (mol wt 44,303). This sequence included the 24-amino acid leader peptide moiety (mol wt 2,576) and 388 amino acids corresponding to the mature protein (mol wt 41,727). The comparison of SCAD and medium chain acyl-CoA dehydrogenase sequences revealed a high degree of homology, suggesting that these enzymes evolved from a common ancestral gene and belong to a gene family. We also studied mutant human SCAD in cultured skin fibroblasts from three patients with hereditary SCAD deficiency. Labeling fibroblast cultures with [35S]-methionine followed by immunoprecipitation with anti-SCAD antibody revealed that a normal size variant SCAD protein was synthesized. In all of the three SCAD-deficient cell lines, the size of variant SCAD mRNA as determined by Northern blotting using one of the normal SCAD cDNA as a probe was also normal, and no difference was observed on Southern blots in the restriction patterns of mutant genomic DNA using EcoRI, TaqI, HincII, and BamHI. These results suggest that the defects in SCAD in these cell lines are caused by a point mutation. PMID- 2565346 TI - Stable remission of tardive dyskinesia by L-dopa. AB - Thirty-five adult chronic schizophrenic patients who had had severe persistent tardive dyskinesia for many years and who had received long-term neuroleptic therapy were treated with small repeated doses of L-dopa. After 4 weeks of treatment the intensity and frequency of involuntary movements decreased, and after 3 months orofacial and choreoathetotic dyskinetic movements diminished very much in all patients. Discontinuation of L-dopa therapy in 10 patients resulted in the return of involuntary movements after 6 weeks. Readministration of the same dose of L-dopa produced the previous therapeutic effects in all patients. Using the NIMH Abnormal Involuntary Movement Scale, the patients were rated severe (4) before treatment and mild (2) after treatment. Maintenance of all 35 patients on daily haloperidol 15 mg, or its neuroleptic equivalent, and small repeated doses of L-dopa induced a stable remission of all involuntary dyskinetic movements for the study year. The placebo control group remained unchanged with the same severe persistent dyskinetic manifestations. PMID- 2565345 TI - The human platelet alloantigens, PlA1 and PlA2, are associated with a leucine33/proline33 amino acid polymorphism in membrane glycoprotein IIIa, and are distinguishable by DNA typing. AB - The human platelet alloantigens, PlA1 and PlA2, comprise a diallelic antigen system located on a component of the platelet fibrinogen receptor, membrane glycoprotein (GP) IIIa. Of the known platelet alloantigens, PlA1, which is carried by 98% of the caucasian population, appears to be the alloantigen that most often provokes neonatal alloimmune thrombocytopenic purpura and posttransfusion purpura. The structural features of the GPIIIa molecule responsible for its antigenicity are as yet unknown. Using the polymerase chain reaction (PcR), we amplified the NH2-terminal region of platelet GPIIIa mRNA derived from PlA1 and PlA2 homozygous individuals. Nucleotide sequence analysis of selected amplified cDNA products revealed a C in equilibrium T polymorphism at base 196 that created a unique Nci I restriction enzyme cleavage site in the PlA2, but not the PlA1 form of GPIIIa cDNA. Subsequent restriction enzyme analysis of cDNAs generated by PcR from 10 PlA1/A1, 5 PlA2/A2, and 3 PlA1/A2 individuals showed that Nci I digestion permitted clear discrimination between the PlA1 and PlA2 alleles of GPIIIa. All PlA2/A2 individuals studied contain a C at base 196, whereas PlA1 homozygotes have a T at this position. This single base change results in a leucine/proline polymorphism at amino acid 33 from the NH2 terminus, and is likely to impart significant differences in the secondary structures of these two allelic forms of the GPIIIa molecule. The ability to perform DNA-typing analysis for PlA phenotype may have a number of useful clinical applications, including fetal testing and determination of the phenotype of severely thrombocytopenic individuals. PMID- 2565347 TI - Neuroleptic-valproic acid combination in treatment of psychotic symptoms: a three case report. AB - Valproic acid is infrequently used for treatment of psychosis. Three consecutive patients with severe neuroleptic-resistant psychotic symptoms responded dramatically to the combination of valproic acid and neuroleptics. All three had normal EEGs and CT scans. Outpatient follow-up showed that the combination was effective in maintaining remissions. PMID- 2565348 TI - Brainstem afferents to the tuberomammillary nucleus in the rat brain with special reference to monoaminergic innervation. AB - Monoaminergic innervation of a histamine-producing cell group, the tuberomammillary nucleus in the posterior hypothalamus, was investigated in the rat by light and electron microscopic immunohistochemical techniques. Immunohistochemical staining of sections of the posterior hypothalamus was demonstrated afferent fibers immunoreactive to tyrosine hydroxylase in ventral and medial subgroups of the tuberomammillary nucleus afferent fibers immunoreactive to tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH), phenyletanolamine-N-methyltransferase (PNMT), and serotonin (5-HT). TH- and DBH immunoreactive fibers were similar and were evenly and densely distributed throughout the tuberomammillary nucleus. Fibers stained with 5-HT antibodies were also present throughout the tuberomammillary nucleus but exhibited the densest labeling in the dendritic layer adjacent to the glia limitans in the ventral subgroup. Innervation by PNMT-immunoreactive axons was sparse. Electron microscopic analysis of TH-, DBH-, and 5-HT-immunoreactive fibers in the tuberomammillary nucleus revealed vesicle-containing terminal boutons, which formed synapses with dendrites of varying size. Synaptic contacts with nerve cell bodies were not found. Retrograde transport of the fluorescent dye Fast Blue injected into the tuberomammillary nucleus, combined with immunofluorescent staining with anti-TH, anti-DBH, anti-PNMT, and anti-5-HT antibodies, showed that monoaminergic input to the tuberomammillary nucleus originated mainly from the adrenergic and noradrenergic cell groups C1-C3 and A1-A2, respectively, and from the serotoninergic cell groups B5-B9 as designated by Dahlstrom and Fuxe ('65). Few double-labeled neurons were found in the nucleus locus coeruleus and the dopaminergic cell groups of the rostral brain stem. The present findings suggest that the activity of the histamine-producing neurons of the tuberomammillary nucleus is influenced by monoaminergic neurons in the ventrolateral and dorsomedial medulla oblongata and the raphe nuclei of the rostral brainstem. PMID- 2565349 TI - Ultrastructural analysis of dynorphin B-immunoreactive cells and terminals in the superficial dorsal horn of the deafferented spinal cord of the rat. AB - Light microscopic studies have demonstrated important differences in the distribution of enkephalin and dynorphin cells and terminals in the dorsal horn. Most importantly, dynorphin neurons are located in regions almost exclusively associated with the transmission and/or control of nociceptive messages (laminae I, IIo, and V); enkephalin neurons, although located in the same regions, are also found in areas involved in the transmission of nonnociceptive messages, e.g., laminae IIi and III. To determine whether there are also differences in the synaptic organization of the two opioid peptides, we have examined the distribution of dynorphin B immunoreactivity at the ultrastructural level. The studies were performed in colchicine-treated rats that underwent dorsal rhizotomy so that the relationship of dynorphin terminals and cells to primary afferent terminals could be established. Dynorphin B-immunoreactive cell bodies and dendrites in laminae I and IIo receive convergent primary and nonprimary afferent input, which suggests that dynorphin neurons receive a small-diameter, nociceptive input. Dynorphin terminals predominantly contain round, agranular vesicles; some terminals also contain a few dense core vesicles. Most dynorphin terminals are presynaptic to unlabelled dendrites; both asymmetric and symmetrical axonal contacts were noted. Dynorphin-immunoreactive boutons are also presynaptic to unlabelled cell bodies and spines. Twenty-nine percent of dynorphin terminals were associated with axonal profiles, including degenerating primary afferent terminals; only rarely could a synaptic density be detected. Although some degenerating primary afferent terminals were clearly presynaptic to dynorphin-immunoreactive terminals, in most cases, the polarity of the relationship between primary afferents and dynorphin terminals could not be established. These data indicate that synaptic interactions made by and with dynorphin-immunoreactive cells and terminals in the superficial dorsal horn are not very different from those that were previously reported for enkephalin cells and terminals. Thus, it is unlikely that dynorphin terminals provide a significant presynaptic input to primary afferent fibers. On the other hand, the presence of a primary afferent input to dynorphin cell bodies and dendrites in the superficial dorsal horn suggests that dynorphin cells receive a direct input from small-diameter, nociceptive primary afferents. That connection might contribute to the increased levels of dynorphin message and peptide that have been reported in rats experiencing a chronic inflammatory condition.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2565350 TI - Nigrotectal projection to the inferior colliculus: horseradish peroxidase transport and tyrosine hydroxylase immunohistochemical studies in rats, cats, and bats. AB - The present study investigated descending projections from the substantia nigra to the auditory tectum. Small (0.02-0.05 microliters) injections of a 30-60% aqueous solution of horseradish peroxidase (HRP) were made unilaterally into the inferior colliculus in rats, cats, and bats (Eptesicus fuscus). Tissue blocks including the substantia nigra, superior colliculus, and inferior colliculus were removed, sectioned, and processed for visualization of HRP. Results show that the substantia nigra, pars lateralis, projects to the inferior colliculus ipsilaterally. In addition, retrogradely labeled cells are found dorsal to the pars lateralis, in a column within the lateral tegmental area of the midbrain. Analysis of injection sites suggests that the principal target of this nigral projection is the dorsal and rostral pericentral region of the inferior colliculus. Immunohistochemical studies with an antibody to tyrosine hydroxylase demonstrate catecholaminergic neurons within the pars lateralis and lateral tegmentum that are similar in location and morphology to one class of HRP retrogradely labeled cells within these structures. These immunohistochemical studies also demonstrate a plexus of fine, varicose tyrosine hydroxylase-positive axons in the rostral pericentral region of the colliculus. The presence of this nigrotectal projection to the inferior colliculus is discussed in relation to its possible role in the control of acousticomotor behavior. PMID- 2565351 TI - Human postnatal CD4- CD8- CD3- thymic T cell precursors differentiate in vitro into T cell receptor delta-bearing cells. AB - The signals required for activation and the differentiation of human triple negative postnatal thymocytes were studied in vitro. Highly purified populations of CD4-, CD8-, CD3- (triple negative) thymocytes were isolated by combined panning and preparative cell sorting and the ability of triple negative thymocytes to proliferate in response to various cytokines determined. Maximal triple negative proliferation was obtained using a mitogenic combination of CD2 antibodies and either rIL-2 or the phorbol ester, PMA. Long term growth (2 to 6 wk) of postnatal triple negative thymocytes was best achieved using CD2 antibodies and rIL-2. After in vitro culture with CD2 antibodies and rIL-2, triple negative thymocytes gave rise to TCR-delta+ cells beginning on day 2 of culture (approximately 15% CD3/TCR-delta+) reaching maximum (approximately 60% CD3/TCR-delta+) on day 7 with stable number of TCR-delta+ cells observed in vitro for up to 6 wk. Analysis of 30 clones of human postnatal triple negative thymocytes demonstrated 9 of 30 (30%) were TCR-delta+, beta F1-, essentially ruling out overgrowth of the triple negative population over time by a minor pool of contaminating TCR-delta+ cells. Thus, these studies have defined an in vitro culture system for human postnatal T cell precursors and demonstrated that precursors of human TCR-gamma delta+ T cells reside in the triple negative thymocyte pool. PMID- 2565352 TI - Distribution and ontogeny of CD2 expression by murine T cells. AB - We have raised a polyclonal antiserum to murine CD2 by immunization of a rabbit with a synthetic peptide corresponding to a hydrophilic sequence in the extracellular domain of the murine CD2 gene. The antiserum immunoprecipitates a 55 kDa protein, consistent with the size predicted by the cDNA sequence. Flow microfluorometric analysis of a panel of T cell tumors and clones demonstrated concordance of reactivity of intact cells with the anti-CD2 serum and the presence of CD2 mRNA. Surprisingly, although splenic T cells were found to uniformly express high levels of CD2, several of a panel of functional T cell clones were found to lack CD2 expression. This suggests that the clones lost CD2 upon in vitro cultivation, and may not be required for activation or maintenance in culture. Adult thymocytes exhibited heterogeneous expression of CD2. The majority of CD4-8- thymocytes expressed low levels and CD4+8+ thymocytes intermediate levels, whereas all CD4+8- and the majority of CD4-8+ thymocytes expressed high levels of CD2. Multiparameter analysis of CD2 expression and that of CD3, CD5, JIId, and IL-2R p55 chain showed that expression of CD2 correlates with the maturational state of thymocytes. Finally, analysis of fetal thymuses from timed pregnancies revealed that expression of CD2 is preceded by that of IL 2R p55 chain. PMID- 2565353 TI - Expression of adhesion structures during B cell development in man. AB - We have used three-color flow cytometry to study the expression of adhesion structures during B cell development in man. The results indicate that the cell surface molecule(s) recognized by 515, a mAb which defines a broadly expressed family of cell-surface glycoproteins that includes lymphocyte homing receptors, exhibit a clear bimodal distribution (515lo and 515hi); 515hi cells were found exclusively on more mature B cells which already expressed high levels of CD20. Earlier, less mature B cells, identified by their expression of CD10, were uniformly 515lo. In contrast, the CD11a/LFA-1 Ag was acquired gradually over the course of B cell development. B cells which expressed high levels of CD20 expressed three to six times as much CD11a/LFA-1 as cells which expressed CD10. Interestingly, expression of the 515hi phenotype was tightly correlated with that of Leu-8, a marker previously shown to define maturational and functions subsets of B cells. These data document the coordinated regulation of multiple cell surface structures during B cell ontogeny, and demonstrate that adhesion structures necessary for proper B cell function are precisely up-regulated during B cell differentiation in man. PMID- 2565354 TI - Accumulation of 111In-neutrophils in rabbit skin in allergic and non-allergic inflammatory reactions in vivo. Inhibition by neutrophil pretreatment in vitro with a monoclonal antibody recognizing the CD18 antigen. AB - The mAb 60.3 recognizes the neutrophil CD18 Ag. We have investigated the effect of in vitro pretreatment of radiolabeled neutrophils with mAb 60.3 on their accumulation in vivo. Further, we have compared the in vivo effects of mAb 60.3 with its effects on neutrophil adherence in vitro. Neutrophil accumulation in vivo was measured in response to: 1) exogenous mediators FMLP, C5a des Arg, LTB4 and IL-1; 2) endogenous mediators generated in a non-allergic inflammatory reaction induced by zymosan; and 3) endogenous mediators generated in two allergic inflammatory reactions, a passive cutaneous anaphylactic reaction and a reversed passive Arthus reaction in rabbit skin. Pretreatment of neutrophils with mAb 60.3 inhibited their accumulation in all the responses. The results demonstrate that there is a common mechanism mediating neutrophil accumulation in these inflammatory reactions. Neutrophils pretreated with mAb 60.3 were also unresponsive to chemoattractants in in vitro adherence assays. However, the antibody-treated neutrophils responded normally to FMLP and C5a with respect to granular enzyme release. These results suggest that the basal expression of CD18 Ag is important for the adherence of neutrophils to microvascular endothelial cells stimulated by the local generation, or administration, of chemical mediators in vivo. Despite the fact that mediators such as FMLP can increase CD18 expression in vitro, it appears more likely that such mediators act in vivo by inducing a conformational change in the basally expressed neutrophil adhesive molecules. PMID- 2565355 TI - Lymphomas with acquired mouse mammary tumor virus proviruses resemble distinct prethymic and intrathymic phenotypes defined in vivo. AB - A number of murine T cell lymphomas expressing the T cell Ag Thy-1 contain acquired mouse mammary tumor (MMTV) proviruses. These lymphomas all express detectable levels of MMTV RNA, yet the majority of the tumors fail to produce MMTV particles. To determine if the ability of lymphomas to produce MMTV is a reflection of the differentiation state of the tumor, we examined eight lymphomas for expression of surface B and T cell Ag as well as for rearrangements and expression of TCR genes. All tumors could be grouped into three categories observed in vivo, including early lymphoid, nonmature intrathymic T cells, and immature intrathymic T cells. Cell lines corresponding to all three phenotypes produced MMTV particles, suggesting that production of virus is not linked to the differentiation state of lymphoid cells. These studies highlight the potential advantage of studying T cell lymphomas vs mixed primary populations or T cell hybridomas for evaluation of both phenotypic and molecular markers in clonal T cells. PMID- 2565356 TI - Effect of Bacillus sphaericus and Bacillus thuringiensis on acid-phosphatase activity of mosquito larvae, Culex pipiens and Aedes caspius. AB - The use of Bacillus sphaericus and B. thuringiensis H-14 form one of the important group of biological control agent against mosquito larvae. Acid phosphatase enzyme plays a significant role in determining susceptibility of mosquito larvae to both bacterial species. Biochemical assay showed activation in acid phosphatase in Culex pipiens and Aedes caspius treated with B. thuringiensis. Variation in acid phosphatase activity occurred in both mosquitoes treated with B. sphaericus, while there was no change in acid phosphatase activity in A. caspius. An obvious increase in activity in C. pipiens treated with the same bacteria was observed. PMID- 2565357 TI - The informal caregiver responsible for home care of the individual with cognitive dysfunction following brain injury. AB - The brain-injured often require care long after they leave the acute care setting. Increasingly this care is being given at home. Informal caregivers are an important link to the brain-injured patient's achieving optimal health and wellness. Characteristics of caregivers are given and the AFFIRM model used as a guide to assist nurses in their preparation of caregivers. A case study illustrates use of the model. PMID- 2565358 TI - Transglutaminase levels and immunologic functions of BCG-elicited mouse peritoneal macrophages isolated by centrifugal elutriation. AB - BCG-elicited mouse peritoneal macrophages were separated into three subpopulations by counterflow centrifugal elutriation. The three subpopulations were characterized on the basis of the level of a protein cross-linking enzyme, tissue transglutaminase. Subpopulation-3 consisted of large cells (greater than 95% esterase positive and greater than 90% viable) and had at least a fivefold higher transglutaminase activity (35 +/- 6 nmol/hr/mg) as compared to macrophages in subpopulation-1 (6 +/- 2 nmol/hr/mg) and at least a threefold higher enzyme activity as compared to subpopulation-2 (11 +/- 2 nmol/hr/mg). Subpopulation-3 also showed sevenfold higher phagocytosis of IgG-coated sheep red blood cells. The three subpopulations showed no difference in their ability to kill Listeria monocytogenes as determined by [3H]-thymidine release. Subpopulations-2 and -3 caused 90% inhibition of murine adenocarcinoma (EMT-6) tumor cell growth in the presence or absence of lipopolysaccharide. Subpopulation-1 had a poor ability to inhibit EMT-6 cell growth (29 +/- 12%). However, in the presence of lipopolysaccharide, this activity increased by at least threefold (92 +/- 7%). The three subpopulations showed no significant difference in their cytolytic activity against murine mastocytoma (P815) target cells in the presence or absence of lipopolysaccharide. These results suggest that tissue transglutaminase may have no significant role in bactericidal, tumoricidal, or tumoristatic function of macrophages; however, it might have some role in promoting the Fc receptor-mediated phagocytic function of the macrophages. PMID- 2565359 TI - The influence of repeated treatment with imipramine, (+)- and (-)-oxaprotiline on behavioural effects of dopamine D-1 and D-2 agonists. AB - The paper examined the action of imipramine, (+)- and (-)-oxaprotiline, administered repeatedly to rats, on the behavioural effects of the dopamine D-1 and D-2 agonists, SKF 38393 and quinpirole, respectively. The three antidepressants studied, given in the single dose or repeatedly, attenuate the enhanced grooming evoked by SKF 38393. The locomotor hyperactivity, evoked by quinpirole administered s.c., is increased by repeated but not single-dose treatment with imipramine and (+)-oxaprotiline [but not with (-)-oxaprotiline]. Quinpirole at a low dose produces the locomotor hypoactivity which is attenuated by repeated, but not single-dose, treatment with the anti-depressants studied here. Repeated imipramine and (+)-oxaprotiline [but not (-)-oxaprotiline] increase the locomotor activity effect of quinpirole injected into the nucleus accumbens. The results indicate that the enhanced responsiveness of the dopamine system, observed previously after repeated treatment with antidepressants, may be mediated by the dopamine D-2 receptors. PMID- 2565360 TI - Dopamine D-2 antagonists reverse apomorphine-induced decreased water intake in the rat: prediction of antipsychotic drugs with few extrapyramidal side-effects? AB - Water intake in water deprived rats was decreased by administration of a low dose of apomorphine (0.1 mg/kg s.c.). This dose is too low to induce hyperactivity and stereotypies. Four different dopamine (DA) D-2 antagonists were used to counteract this effect of apomorphine; haloperidol [an antipsychotic inducing extrapyramidal side-effects (EPS)], sulpiride (an antipsychotic inducing less EPS than haloperidol), metoclopramide (not used as an antipsychotic but inducing EPS) and domperidone (not passing through the blood brain barrier). Domperidone did not counteract the apomorphine effect, indicating a central mechanism of action for apomorphine. Metoclopramide did not counteract the apomorphine effect and, in higher doses, water intake was even further reduced. Sulpiride completely counteracted the apomorphine effect but, in higher doses, did not by itself reduce water intake. Haloperidol counteracted the apomorphine effect in a small dose-range and caused a further reduction in the water intake when given in high doses. The results can be explained by the existence of two subpopulations of D-2 receptors related to different functions. The model described may be used in screening experiments aimed at finding new antipsychotic drugs with a low incidence of EPS. PMID- 2565362 TI - Production and maintenance of large numbers of Dugesia tigrina (Turbellaria: Tricladida) for the control of mosquitoes in the field. AB - Methods to increase and preserve Dugesia tigrina for mosquito control purposes through cocoon production, mechanical sectioning and cold storage were examined. Cocoon production was an ineffective and unreliable means of increasing planarian numbers, with an average of only 0.19 cocoons containing 0.74 young produced per adult per week. In comparison, mechanical sectioning proved to be more appropriate for increasing the number of planaria. One hundred planaria averaging 8 mm in length could be sectioned into 600 segments in approximately 18 min; regeneration was achieved by approximately 94% of these segments, usually within 8 days after sectioning. Techniques were developed to store planaria at 10 degrees C until needed for field release. PMID- 2565361 TI - A metabolite of buspirone increases locus coeruleus activity via alpha 2-receptor blockade. AB - Extracellular single unit recording techniques was used to pharmacologically analyze the excitatory action of buspirone on locus coeruleus (LC) noradrenergic neurons. Intravenously administered buspirone (0.5-8 mg/kg) dose-dependently increased LC firing rate. Furthermore, pretreatment with buspirone (8 mg/kg, i.p.) caused a parallel shift to the right of the dose-response curve for the inhibitory action of the alpha 2-receptor agonist clonidine on LC neurons. The inhibitory effect of microiontophoretically applied noradrenaline on LC neurons was not altered by the simultaneous application of buspirone, but almost totally blocked by its major metabolite 1-(2-pyrimidinyl-piperazine) (1-PP). The results indicate that buspirone causes activation of LC neurons via an alpha 2-receptor antagonistic action of its metabolite, 1-PP. PMID- 2565363 TI - Impact of new insect growth regulators and their formulations on mosquito larval development in impoundment and floodwater habitats. AB - Four insect growth regulators were evaluated in the laboratory and field. In the laboratory, AC-291898 showed excellent activity, inducing 90% mortality in Culex quinquefasciatus and Aedes aegypti at 0.5-0.7 ppb. The EC formulation of XRD-473 was slightly more active than the technical material with an LC90 of 0.84 ppb and 0.92 ppb against Ae. aegypti and Cx. quinquefasciatus, respectively. In the field, AC-291898 at the rates of 0.005 and 0.01 lb AI/acre caused 85 and 100% inhibition of adult emergence of Cx. tarsalis 2 days after treatment. Activity, however, declined at the low rate, while the high rate remained active for more than 7 days. In the same species, XRD-473 induced complete inhibition of adult emergence 2 days after treatment at the rates of 0.01, 0.025 and 0.05 lb AI/acre. At the low rate (0.01 lb AI/acre), activity declined markedly, while the 2 higher rates remained active for one week. In Cx. peus larvae, AC-291898 at the rates of 0.005, 0.01 and 0.025 lb AI/acre produced complete inhibition of adult emergence 2 days after treatment, but activity declined at the 2 lower rates 7 days after treatment, while the high rate (0.025 lb AI/acre) remained active for more than one week. Methoprene (4%) pellets were effective against Cx. tarsalis for 7 days at the rates of 0.25 lb AI/acre whereas 0.5 lb AI/acre was required to obtain similar results against Cx. peus larvae.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565364 TI - New mosquito records for Nepal. AB - Thirteen mosquito species of the genera Aedes, Culex, Mimoyia and Uranotaenia are reported from Nepal for the first time. Taking into account taxonomic changes, 70 species of culicine (i.e., non-anopheline) mosquitoes are now known from Nepal. PMID- 2565365 TI - Diurnal pattern and behavior of oviposition of Toxorhynchites theobaldi in the field. AB - The diurnal pattern and oviposition behavior of Toxorhynchites theobaldi natural populations were studied in 25 artificial containers in the field. The mosquito exhibited a bimodal oviposition pattern with the lower peak at 1100 hr and a mean of 15.7 eggs per container. The higher peak was observed at 1900 hr with a mean of 80.9 eggs per container. Each female flew from 21 to 58 elliptic vertical circles before ejecting one egg upon the surface. In 270 oviposition events, the average was 31.4 ellipses, and the frequency distribution of flights number with different ellipse numbers was fitted to a Poisson distribution. There was a significant linear correlation (r = 0.70) between the oviposition rate and the container surface area. PMID- 2565366 TI - Significant associations between mosquito control service requests and mosquito populations. AB - Correlation and multiple regression analyses were used to examine the relationships between mosquito population densities and the numbers of telephone requests for mosquito control services made by people in Polk County, Florida. In 32 of 42 census tracts, there were significant (P less than 0.05) correlations between mosquitoes and requests and that, for 16 tracts, a significant linear relationship could be demonstrated between service requests and only one mosquito taxon. The significant associations were dependent on the types of mosquitoes rather than the relative abundance among mosquito groups. Service requests were most often related to Aedes, followed in importance by Culex salinarius and Anopheles; least often to Cx. nigripalpus. Evidence to support a cause and effect relationship is also presented. PMID- 2565367 TI - Evaporation and skin penetration characteristics of mosquito repellent formulations. AB - Formulations of the mosquito repellent N,N-diethyl-3-methylbenzamide (deet) in combination with a variety of additives were developed to control repellent evaporation and percutaneous penetration. Deet was also formulated with the repellent dimethyl phthalate to study the interaction of the two compounds on the skin. The evaporation and penetration processes were evaluated on whole and split thickness pig skin using radiolabeled repellents with an in vitro apparatus. Under essentially still air and air flow conditions, one of the deet formulations resulted in significantly reduced total evaporation and percutaneous penetration of deet as compared to unformulated repellent. When deet and dimethyl phthalate were combined, neither repellent affected the total amount of evaporation and penetration of the other compound. However, initial percutaneous penetration and evaporation rates were slightly less and decayed less rapidly than when both chemicals were tested separately at the same dose. These results indicated a degree of competition of the two compounds for the same avenues of loss. PMID- 2565368 TI - Laboratory evaluation of controlled-release repellent formulations on human volunteers under three climatic regimens. AB - Six controlled-release personal-use topical insect/arthropod repellent formulations of diethylmethylbenzamide (deet) were evaluated in an environmental chamber on volunteers for repellency against the mosquitoes Aedes aegypti and Ae. taeniorhynchus under three climatic regimens: basic variable high humidity (tropical environment), basic constant high humidity (forested and wet environment) and basic hot (hot-dry environment). The best protection under all the climatic regimens was provided by the Biotek formulation. In a tropical environment, some formulations induced more biting from mosquitoes than the concurrent untreated control in the late hours of the testing. Repellency was not directly related to the deet concentration in the various controlled-release repellent formulations. PMID- 2565369 TI - Effects of release rates on the range of attraction of carbon dioxide to some southwestern Ontario mosquito species. AB - The effects of release rates of 0, 250, 500, 1,000 and 4,000 ml/min on the range of attraction of carbon dioxide to some southwestern Ontario mosquito species was determined using ramp traps placed at 3, 7, 11, 15 and 19 m from a central pressurized cylinder. For female Aedes vexans, spring Aedes spp. and Anopheles walkeri, an increase in the release rate of CO2 from 1,000 to 4,000 ml/min resulted in extension of the range of attractiveness from between 3-7 m to between 7-11 m; rates of 500 and 250 ml/min did result in an increase in number of mosquitoes in the traps. Correspondingly, significantly more mosquitoes were caught in the traps at 3 m when the rate was increased to 1,000 ml/min from 500 ml/min. For Ae. vexans, 4,000 ml/min of CO2 attracted more mosquitoes to the 7 m traps than 1,000 ml/min. In this work carbon dioxide did not result in an increase in the number of Culiseta inornata, Cs. morsitans and Culex restuans and Cx. pipiens in the traps. PMID- 2565370 TI - A self-marking device for emergent adult mosquitoes. AB - A portable, tent-like device with powdered fluorescent pigment for marking emergent adult mosquitoes was evaluated in the field using Culex quinquefasciatus adults. The top of the device was equipped with cheesecloth partitions impregnated with the pigment. Eighty-six percent of the emergent adults passed through the partitions, and all were marked. PMID- 2565371 TI - Changes in excitatory amino acid receptor binding in the intact and decorticated rat neostriatum following insulin-induced hypoglycemia. AB - An involvement of excitatory amino acid (EAA) transmitter-receptor interactions in the development of hypoglycemia-induced neuronal damage has been suggested. We report here on the binding to EAA receptors in the rat caudate nucleus and cerebral cortex, during and following severe insulin-induced hypoglycemia with an isoelectric EEG of 10 or 30 min duration. The binding of alpha-[3H]amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid [( 3H]AMPA) to quisqualate receptors, [3H]kainic acid (KA) to kainate receptors, and [3H]glutamate to N-methyl-D aspartate (NMDA)-sensitive sites was determined by quantitative autoradiography. During EEG isoelectricity, AMPA binding was reduced by approximately 40%, which could represent quisqualate receptor desensitization. One hour following glucose induced recovery, AMPA binding was no longer different from control level. As the recovery period was prolonged to 1 or 4 weeks, AMPA binding decreased. The decrease was more pronounced in the dorsolateral than in the ventromedial part of the striatum. This correlates with the distribution of neuronal damage, and probably reflects loss of receptor binding sites due to cell death. During the period of EEG silence there was a tendency toward an increase in NMDA displaceable glutamate binding. Following 4 weeks of recovery, binding to NMDA receptors was significantly decreased. Glutamate binding to NMDA-sensitive sites was remarkably resistant to neuronal necrosis and was not significantly different from control values in the dorsolateral caudate 1 week following the hypoglycemic coma. No changes in KA binding were found until 1 week posthypoglycemia, when a significant reduction in binding was noted in the lateral striatum.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565372 TI - Delayed neurologic deterioration following anoxia: brain mitochondrial and metabolic correlates. AB - Hyper- but not normoglycemic cats exposed to 8 min of anoxia show neurologic signs (fasciculations, myoclonic jerks, seizures) that develop after a symptom free period. We examined brain mitochondrial function and metabolite concentrations at 0, 1, 3, and 5 h following exposure to anoxia, to correlate biochemical findings with the presence ("symptomatic") or absence ("presymptomatic") of neurologic signs. Brain mitochondria isolated postexposure only from symptomatic cats showed markedly decreased (-50%), state 3 (ADP stimulated), and uncoupler-stimulated respiration rates with NAD- and FAD-linked substrates. Respiratory control and ADP/oxygen (ADP/O) ratios remained unchanged, indicating, respectively, that coupling and efficiency of ATP synthesis were preserved. Thus, inhibition of electron transport chain function, not phosphorylative activity, may be rate limiting for respiration. During anoxia, hyperglycemic cats showed higher brain lactate levels (26 versus 20 mumol/g), but similar ATP and phosphocreatine concentrations, compared with normoglycemic cats. After exposure, in all animals lactate and phosphocreatine were restored to control levels, whereas ATP remained at 85%. Cats that became symptomatic demonstrated four- to sixfold increases in lactate and 50% reductions in phosphocreatine. At 3 and 5 h postexposure, symptomatic animals showed significant reductions in ATP concentrations. We conclude that although initially asymptomatic, hyperglycemic cats exposed to anoxia undergo a neurologic deterioration over several hours following reoxygenation that is correlated with inhibition of mitochondrial respiration, increases in tissue lactate, and decreases in energy state. PMID- 2565373 TI - Prenatal ontogenesis of brain phenolamines and catecholamines in relation to their metabolizing enzymes in Roman avoider strains of rats. AB - Phenolamines, particularly octopamines, are of special importance in avoidance behavior. In the Roman low avoidance (RLA) strain, p-octopamine can induce locomotor behavioral activity that is normally observed in the Roman high avoidance (RHA) strain. For these reasons, the levels of prenatal octopamines (para and meta isomers) have been studied in relation to noradrenaline and dopamine levels. In the hypothalamus and brainstem of RHA, a maximum level of the para isomer is observed at 15 days of embryonic development but, unlike in controls and RLA animals, this level remains almost constant until 20 days. For the meta-isomer and catecholamines, there is a 1-2 day delay in detection between controls and RLA or RHA. The study of related enzyme activities reveals that tyrosine hydroxylase displays a 2-day delay in RHA when compared to the control value at 19 days of fetal life. These results are discussed in terms of the role of p-octopamine in avoidance conditioning and of the possible delayed expression of the tyrosine hydroxylase gene in Roman strains of rats. PMID- 2565374 TI - Solubilization of quisqualate-sensitive [3H]glutamate binding activity from rat retina. AB - Binding activity of a putative central neurotransmitter, L-glutamic acid, was examined in the supernatant preparations solubilized from rat retinal membranes by Nonidet P-40. [3H]Glutamate binding activity increased linearly with increasing concentrations of the solubilized proteins up to 15 micrograms. The binding activity reached an equilibrium within 10 min at 2 degrees C, while increasing with incubation time up to 60 min at 30 degrees C. Addition of an excess of nonradioactive glutamate rapidly decreased the activity at 30 degrees C. Scatchard analysis revealed that the solubilized retinal binding activity consisted of a single component with a KD of 0.25 microM and a Bmax of 57.4 pmol/mg protein. The solubilized binding activity exhibited a stereospecificity and a structure selectivity to L-glutamate, and was abolished by quisqualate, L glutamate diethyl ester, and DL-2-amino-3-phosphonopropionate. None of the other agonists and antagonists for the central excitatory amino acid receptors affected the binding activity. Reduction of incubation temperature from 30 degrees C to 2 degrees C resulted in a drastic attenuation of the binding activity due to decrement of the number of the apparent binding sites. Cation-exchange column chromatography revealed that unidentified radioactive material was in fact formed during the incubation of [3H]glutamate with the retinal preparations at 30 degrees C. These results suggest that retinal [3H]glutamate binding activity may be derived at least in part from the quisqualate-sensitive membranous enzyme with a stereospecific and structure-selective high affinity for the central neurotransmitter. PMID- 2565375 TI - Dopamine autoreceptors modulate the phosphorylation of tyrosine hydroxylase in rat striatal slices. AB - The hypothesis that dopamine (DA) autoreceptors modulate the phosphorylation of tyrosine hydroxylase (TH; EC 1.14.16.2) was investigated in rat striatal slices. Tissue was prelabeled with 32P inorganic phosphate, and TH recovered by immunoprecipitation with anti-TH rabbit serum. The TH monomer was resolved on sodium dodecyl sulfate polyacrylamide gels, and the extent of phosphorylation was determined by scanning densitometry of autoradiographs. Depolarization of striatal slices with 55 mM K+ markedly increased the incorporation of 32P into several proteins, including the TH monomer (Mr = 60,000). A similar increase in TH phosphorylation occurred in response to the adenylate cyclase activator forskolin and the cyclic AMP analog dibutyryl cyclic AMP. An increase in TH phosphorylation was not observed in response to the D1-selective agonist SKF 38393. The D2-selective DA autoreceptor agonist pergolide decreased the phosphorylation of TH below basal levels and blocked the increase in phosphorylation elicited by 55 mM K+. The inhibitory effect of pergolide was antagonized by the D2-selective antagonist eticlopride. Changes observed in the phosphorylation of TH were mirrored by changes in tyrosine hydroxylation in situ. These observations support the hypothesis that a reduction in TH phosphorylation is the mechanism by which DA autoreceptors modulate tyrosine hydroxylation in nigrostriatal nerve terminals. PMID- 2565376 TI - Effect of impact trauma on neurotransmitter and nonneurotransmitter amino acids in rat spinal cord. AB - N-Methyl-D-aspartate (NMDA) administration exacerbates neurological dysfunction after traumatic spinal cord injury in rats, whereas NMDA antagonists improve outcome in this model. These observations suggest that release of excitatory amino acids contributes to secondary tissue damage after traumatic spinal cord injury. To further examine this hypothesis, concentrations of free amino acids were measured in spinal cord samples from anesthetized rats subjected to various degrees of impact trauma to the T9 spinal segment. Levels of excitatory and inhibitory neurotransmitter amino acids [gamma-aminobutyric acid (GABA), glutamate, aspartate, glycine, taurine] and levels of nonneurotransmitter amino acids (asparagine, glutamine, alanine, threonine, serine) were determined at 5 min, 4 h, and 24 h posttrauma. Uninjured surgical (laminectomy) control animals showed modest but significant declines in aspartate and glutamate levels, but not in other amino acids, at all time points. In injured animals, the excitatory amino acids glutamate and aspartate were significantly decreased by 5 min posttrauma, and remained depressed at 4 h and 24 h as compared with corresponding laminectomy controls. In contrast, the inhibitory amino acids, glycine, GABA, and taurine, were decreased at 5 min postinjury, had partially recovered at 4 h, and were almost fully recovered at 24 h. The nonneurotransmitter amino acids were unchanged at 5 min posttrauma and significantly increased at 4 h, with partial recovery at 24 h. At 4 h postinjury, severe trauma caused significantly greater decreases in aspartate and glutamate than did either mild or moderate injury. These findings are consistent with the postulated role of excitatory amino acids in CNS trauma. PMID- 2565377 TI - Tyrosine hydroxylase and levodopa responsive dystonia. AB - It has been suggested that a form of inherited dystonia responsive to levodopa might be due to an abnormality of tyrosine hydroxylase gene. This hypothesis has been tested using a cDNA tyrosine hydroxylase gene probe in three families with this disorder. No evidence for genetic linkage between the disease and tyrosine hydroxylase loci was found; it is possible that the disorder results from a post transcriptional defect confined to the brain. PMID- 2565378 TI - Dual labelling of circulating CD8 cells in patients with multiple sclerosis. AB - Peripheral blood T cell phenotypes have been analysed in serial samples from patients with multiple sclerosis, their unaffected relatives and controls using a panel of antibodies chosen to distinguish T suppressor and activated suppressor cells from other CD8 lymphocytes. Overall, the percentage of Leu 2a cells correlated with alterations in the Leu 2a/15 suppressor sub-population (r = 0.79, p less than 0.001). Fewer circulating Leu 2a and Leu 2a/15 positive cells were identified in multiple sclerosis patients than unaffected individuals but there was no alteration in percentage of activated (Leu 2a/DR) CD8 cells. These findings suggest that the fluctuations in CD8 cells, characteristic of patients with multiple sclerosis, are due to alterations in T suppressor phenotype; this may then lower the threshold for activation of other T cell subpopulations. PMID- 2565379 TI - EPSPs in rat neocortical neurons in vitro. II. Involvement of N-methyl-D aspartate receptors in the generation of EPSPs. AB - 1. Intracellular recordings were obtained from neurons in layer II/III of rat frontal cortex. Single-electrode current- and voltage-clamp techniques were employed to compare the sensitivity of excitatory postsynaptic potentials (EPSPs) and iontophoretically evoked responses to N-methyl-D-aspartate (NMDA) to the selective NMDA antagonist D-2-amino-5-phosphonovaleric acid (D-2-APV). The voltage dependence of the amplitudes of the EPSPs before and after pharmacologic changes in the neuron's current-voltage relationship was also examined. 2. NMDA depolarized the membrane potential, increased the neuron's apparent input resistance (RN), and evoked bursts of action potentials. The NMDA-induced membrane current (INMDA) gradually increased with depolarization from -80 to -40 mV. The relationship between INMDA and membrane potential displayed a region of negative slope conductance in the potential range between -70 and -40 mV which was sufficient to explain the apparent increase in RN and the burst discharges during the NMDA-induced depolarization. 3. Short-latency EPSPs (eEPSPs) were evoked by low-intensity electrical stimulation of cortical layer IV. Changes in the eEPSP waveform following membrane depolarization and hyperpolarization resembled those of NMDA-mediated responses. However, the eEPSP was insensitive to D-2-APV applied at concentrations (up to 20 microM) that blocked NMDA responses. 4. EPSPs with latencies between 10 and 40 ms [late EPSPs (lEPSPs)] were evoked by electrical stimulation using intensities just subthreshold to the activation of IPSPs. The amplitude of the lEPSP increased with hyperpolarization and decreased with depolarization. 5. The lidocaine derivative QX-314, injected intracellularly, suppressed sodium-dependent action potentials and depolarizing inward rectification. Simultaneously, the amplitude of the eEPSP significantly decreased with depolarization. Neither the amplitude of a long-latency EPSP nor the amplitude of inhibitory postsynaptic potentials (IPSPs) was significantly affected by QX-314. 6. Cesium ions (0.5-2.0 mM) added to the bathing solution reduced or blocked hyperpolarizing inward rectification. Under these conditions, the amplitude of the eEPSP increased with hyperpolarization. The amplitude of the lEPSP was unaltered or enhanced. 7. The lEPSP was reversibly blocked by D-2-APV (5-20 microM), although the voltage-dependence of its amplitude did not resemble the action of NMDA on neocortical neurons.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2565380 TI - Oral 5-aminosalicyclic acid in children with colonic chronic inflammatory bowel disease: clinical and pharmacokinetic experience. AB - Five children between the ages of 10 and 17 years with chronic inflammatory bowel disease with involvement of colon had intolerance to sulfasalazine, and desensitization trials had failed in them. In addition, they were steroid dependent. Therefore treatment was tried with an oral 5-aminosalicylic acid (5 ASA) preparation coated with the pH sensitive polymer Asacol. Three of these patients responded to the new drug and were weaned off the steroids. However, the other two patients developed repeated side effects from the medication. Plasma levels and urinary excretion of 5-ASA and its major metabolite in four of these patients were determined and were noted to be similar to those in the adults. Range of variable steady state plasma level of 5-ASA was 0.1-5 mg/L and of its acetyl-metabolite (Ac-5-ASA) was 1-8 mg/L. PMID- 2565381 TI - Does proctosigmoiditis in inflammatory bowel disease presage the imminent onset of symptoms? AB - This study was undertaken to determine if asymptomatic children and adolescents with inflammatory bowel disease and moderate to severe anorectosigmoid inflammation might remain symptom-free for at least 12 months without specific intrarectal therapy. We prospectively studied 13 asymptomatic patients 6-21 years of age (four with Crohn's disease and nine with nonspecific colitis) with previously documented anorectosigmoid inflammation. Of these 13, four had moderate to severe anorectosigmoid inflammation both endoscopically and histologically. These four patients (two with Crohn's disease and two with nonspecific colitis) were entered into the second phase of the study. Three were receiving sulfasalazine, and one received methylprednisolone, 4 mg/day, and 6 mercaptopurine, 50 mg/day. None received intrarectal therapy. Clinical evaluation revealed that all four remained asymptomatic for 12 months despite the continued presence of moderate to severe anorectosigmoid inflammation. These results indicate that in children and adolescents with inflammatory bowel disease, the presence of inflammation of the anorectosigmoid does not necessarily correlate with or presage the onset of symptoms of proctosigmoiditis. Therefore, active inflammation of the anorectosigmoid is not the sole prerequisite for intrarectal therapy. The clinician should be guided by the symptoms of the patient, not by the presence or absence of active anorectosigmoid inflammation. PMID- 2565382 TI - Recurrent infantile vomiting due to intentional ipecac poisoning. AB - Recurrent vomiting is a common symptom in infancy that, when severe, may prompt an extensive diagnostic evaluation. We report a 1-year-old infant whose recurrent vomiting eluded diagnosis until ipecac syrup was detected in vomitus and urine. Separation of the infant from the mother resulted in complete resolution of the symptoms. In Munchausen syndrome by proxy, a disorder in which a mother fabricates or induces illness in her child, most reported fatalities are due to intentional poisoning. In atypical cases of recurrent vomiting, early screening of body fluids for unexpected drugs, particularly ipecac, may prevent potentially fatal delays in diagnosis. PMID- 2565383 TI - Characterization of the solution degradation products of etintidine, an H2 receptor antagonist. AB - The primary solution degradation products of the antiulcer drug etintidine (1, N" cyano-N-[2-[[(5-methyl-1H-imidazol-4-yl)methyl]thio]ethyl]- N'-2 propynylguanidine) were determined to be the guanyl urea 2, the guanidine 3, the amine 4a, and the cyanoamine 4b. These products resulted from nitrile hydrolysis and/or intramolecular cyclization of the guanidino and propargyl groups. The amine 4a was found to be a predominant degradation product in aqueous media of pH 4-7 at 70 +/- 0.2 degrees C. PMID- 2565384 TI - Acute effects of ranitidine, famotidine and omeprazole on plasma gastrin in the rat. AB - In the rat, treatment with gastric inhibitory drugs may result in hypergastrinemia, an effect thought to be in response to increased gastric pH caused by inhibition of acid secretion. This study compared 24-hr profiles of plasma gastrin levels associated with three different compounds at equivalent, highly effective antisecretory doses. Ranitidine, famotidine and omeprazole at 60, 20 and 40 mg/kg p.o., respectively, inhibited basal acid secretion of chronic gastric fistula rats by greater than 95% and raised intraluminal pH to above 7.0 for 5 hr. The peak plasma gastrin levels associated with each agent were observed 5 hr after dosing. Ranitidine, famotidine and omeprazole induced statistically significant and distinct peak hypergastrinemic responses of 312 +/- 20, 483 +/- 28 and 616 +/- 27 pg/ml, respectively. After 8 hr ranitidine and famotidine associated gastrin values returned to control levels, whereas those of omeprazole remained substantially above control values until the 12th hr. Differences in peak gastrin levels between compounds disappeared at increased dose levels of 500 mg/kg for ranitidine, 200 or 2000 mg/kg for famotidine and 140 mg/kg for omeprazole. Unlike high dose famotidine, omeprazole (140 mg/kg) maintained peak plasma gastrin levels at 8, 12, and 16 hr after dosing. These studies demonstrate clearly hypergastrinemic responses to single dose administration of ranitidine, famotidine and omeprazole. The differences observed in peak plasma gastrin levels at equivalent antisecretory doses of these agents suggests the presence of luminal acid independent components that effect gastrin release. Moreover, these studies indicate that, in the rat, the most unique aspect of omeprazole associated hypergastrinemia is the magnitude of its prolonged response. PMID- 2565385 TI - Kinetics of drug action in disease states. XXIX. Effect of experimental nephrotic syndrome on the pharmacodynamics of heptabarbital: implications of severe hypoalbuminemia. AB - The purpose of this investigation was to determine the effect of nephrotic syndrome (NS) on the pharmacodynamics of a barbiturate. NS was induced in male rats by puromycin aminonucleoside; it caused hypoproteinemia, increased liver and kidney weight and elevated serum creatinine and urea nitrogen concentrations. Serum albumin concentration decreased from 3.5% in controls to 0.90% in NS animals. The rats were infused i.v. with heptabarbital, 1 mg/min, until they lost their righting reflex. The total dose (mean +/- S.D.) required by rats with NS, 40.2 +/- 4.2 mg/kg, was substantially lower than that required by normal animals (68.6 +/- 6.2 mg/kg, P less than .001). Serum protein binding of heptabarbital was reduced from 49% in controls to 26% in NS rats. However, the drug concentration in cerebrospinal fluid (CSF) at the pharmacologic endpoint was not significantly different in controls and NS rats (18.9 +/- 1.5 vs. 18.3 +/- 1.4 mg/l). Serum, CSF and the brain contained appreciable concentrations of a metabolite of heptabarbital. To determine if the metabolite contributes to the pharmacologic effect of the parent drug, rats received an i.v. injection of 46, 60 or 100 mg/kg of heptabarbital. Concentrations of heptabarbital in CSF at return of righting reflex (which occurred after 15, 25 and 50 min, respectively) were independent of dose whereas metabolite concentrations increased with increasing dose. Thus, the metabolite of heptabarbital in male rats is pharmacologically inactive.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565386 TI - Interaction of non-guanylate cyclase-linked atriopeptin receptor ligand and endopeptidase inhibitor in conscious rats. AB - We examined the interaction of SC-46542 [des(Phe106, Gly107, Ala115, Gln116) AP(103-126)], a non-guanylate cyclase-linked atriopeptin (AP) binding site ligand, with thiorphan, an inhibitor of endopeptidase 24.11, on mean arterial pressure, urine flow, urinary sodium excretion and plasma AP immunoreactivity in conscious rats. The coadministration of SC-46542 (16 micrograms/kg/min) and thiorphan (30 mg/kg i.v. bolus) produced a greater diuresis and natriuresis (but had no effect on mean arterial pressure) than administration of either compound alone; plasma APir increased 2-fold during coadministration of SC-46542 and thiorphan (from 325 +/- 46 to 676 +/- 86 pg/ml). Administration of SC-46542 or thiorphan alone had small or no effects on mean arterial pressure, urine flow, urinary sodium excretion or plasma APir. Converting enzyme inhibition did not contribute to the effects of thiorphan since coadministration of captopril plus SC-46542 produced effects similar to SC-46542 alone. When a near threshold infusion of AP(103-126) was combined with the coadministration of SC-46542 and thiorphan, there was a potentiation of the depressor, diuretic and natriuretic responses. Neither SC-46542 nor thiorphan alone had these effects. SC-46542 potentiated the depressor but not diuretic or natriuretic responses to low dose AP(103-126) infusion; thiorphan had little or no effect on the responses to low dose AP(103-126). We conclude that blockade of non-guanylate cyclase-linked AP binding sites with SC-46542 combined with inhibition of AP degradation by endopeptidase 24.11 with thiorphan increases diuresis and natriuresis more than inhibition of either system alone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565387 TI - Acute epinephrine treatment fails to facilitate stimulus-induced neurotransmitter overflow in the isolated perfused rat kidney. AB - The effects of acute in vitro epinephrine treatment (40 nM, 1 hr) on stimulus induced neurotransmitter overflow from the isolated perfused kidney were examined. This treatment resulted in the release of both norepinephrine and epinephrine upon nerve stimulation but no increase in fractional stimulus-induced overflow. Experiments utilizing beta and/or alpha adrenoceptor blockade revealed that beta adrenoreceptor blockade alone had no effect on overflow but that a dose dependent decrease in stimulus-induced overflow was produced by beta adrenoceptor blockade after alpha adrenoceptor blockade. Thus, under normal conditions the facilitatory influence of epinephrine via prejunctional beta adrenoceptors is masked by the prejunctional alpha adrenoceptor negative feedback loop. The effects of extraneuronal presentation of isoproterenol, salbutamol and epinephrine on stimulus-induced neurotransmitter overflow also were examined. In the absence of alpha adrenoceptor blockade, only epinephrine failed to enhance neurotransmitter overflow. After competitive alpha adrenoceptor blockade, all three agents produced significant increases in stimulus-induced neurotransmitter overflow from the rat kidney, although alpha adrenoceptor stimulation appeared to limit this effect at high agonist concentrations. We conclude that the dominance of prejunctional alpha adrenoceptors in the rat kidney results in the inability of epinephrine to produce beta adrenoceptor-mediated facilitation of neurotransmitter overflow even when it is incorporated into renal neurotransmitter stores and coreleased with endogenous norepinephrine upon nerve stimulation. PMID- 2565388 TI - Regulation of Ca++ influx into striatal neurons by kainic acid. AB - We investigated the mechanisms by which kainic acid (KA) produces increases in [Ca++]i in single striatal neurons in vitro using fura-2-based microfluorimetry. When neurons were depolarized by perfusion with high K+ or veratridine containing solutions, [Ca++]i rose rapidly to a peak and then declined to a lower sustained plateau that persisted as long as the depolarizing stimulus. The peak high K+ induced rise in [Ca++]i occurred at [K+]o greater than 50 mM and the plateau was largest at 30 mM K+. [K+]o that was greater than 70 mM caused the magnitude of the plateau to decrease. Responses to high K+ stimulation were completely dependent on [Ca++]o and presumably represented Ca++ influx. Nitrendipine partially blocked the peak of the high K+-induced response and completely blocked the sustained plateau Ca++ influx. The nitrendipine-resistant portion of the high K+ response could be completely blocked by predepolarization of the cell in Ca++ free solution. KA also produced large increases in [Ca++]i that were abolished on removal of external Ca++. Predepolarization/nitrendipine greatly reduced the effect of lower [KA] (100 microM). However, KA-induced increases in [Ca++]i became increasingly resistant to block of voltage-sensitive Ca++ channels as [KA] rose above 100 microM, indicating a second route of Ca++ entry that may be the KA receptor-gated ionophore. About one-half the responses to KA (100 microM) also displayed a large oscillation. [Ca++]i rose to a peak, fell and then rose again before finally declining to a plateau level. This oscillation was abolished when all external Na+ was replaced by Li+ and may result from alterations in the buffering of [Ca++]i as a result of KA-induced Na+ influx. PMID- 2565389 TI - Antidiarrheal activity of alpha-2 adrenoceptor agonist SK&F 35886. AB - Alpha-2 adrenoceptor agonists exhibit antidiarrheal activity in animal models and in humans. However, hypotensive and sedative side effects seriously limit the use of these agents to treat diarrhea. SK&F 35886 (2,6-dimethyl phenylamino imidazoline) is an alpha-2 adrenoceptor agonist with little central nervous system activity. In Ussing chamber preparations of rabbit ileum, SK&F 35886 produces a concentration-dependent decrease in basal short-circuit current (Isc) (EC50 0.2 microM) that is dependent on the presence of mucosal HCO3. This concentration-response curve is shifted to the right of rauwolscine, increasing the EC50 to 30 microM. Prazosin had no effect on this response. Flux studies indicate that SK&F 35886 increases net Cl absorption and enhances HCO3 absorption without altering net Na flux. After PGE2 stimulation of Isc, SK&F 35886, applied either serosally or mucosally, immediately returns the Isc to base line. This effect is due to a reversal of the PGE2-induced inhibition of Na and Cl absorption. In vivo SK&F 35886 dose-dependently inhibits PGE2-induced enteropooling when given orally (ED50 approximately 31 micrograms/kg). This effect is attenuated significantly by rauwolscine (1.0 micrograms/kg s.c.). In cecectomized rats, SK&F 35886 abolishes PGE2-induced diarrhea within 1 hr after oral administration of the drug. SK&F 35886 (500 micrograms/kg p.o.) did not alter hexobarbital sleep time or elicit piloerection or lethargy, whereas clonidine (37.3 micrograms/kg p.o.) significantly enhanced hexobarbital sleep time. These results illustrate the ability of a peripheral acting alpha-2 agonist to promote absorption and inhibit secretion and diarrhea in the mammalian intestine. PMID- 2565390 TI - Effect of gepirone and ipsapirone on the stimulated and unstimulated secretion of prolactin in the rat. AB - Previous studies have demonstrated that the 5-hydroxytryptamine1A (5-HT1A) agonist buspirone stimulated rat prolactin (PRL) secretion. Administration of the 5-HT1A agonists gepirone (GEP) or ipsapirone (IPS) s.c. in doses from 1 to 10 mg/kg had no effect on PRL secretion in male rats. However, pretreatment with GEP (10 mg/kg) or IPS (10 mg/kg) significantly attenuated the increase in serum PRL concentration elicited by the 5-HT agonists 5-methoxy-N,N-dimethyltryptamine or 6 chloro-2-(1-piperazinyl)-pyrazine (MK-212). To determine whether the inhibitory effect of GEP or IPS was related to a serotonergic mechanism or a more general inhibitory effect on PRL secretion, the ability of GEP and IPS to inhibit the haloperidol- or alpha-methyl-p-tyrosine-induced increase in PRL secretion was studied. GEP (1, 3 and 10 mg/kg) and IPS (10 mg/kg) inhibited the increase in PRL secretion produced by either haloperidol (0.25 mg/kg) or alpha-methyl-p-tyrosine (75 mg/kg). Furthermore, GEP produced a concentration-dependent inhibition of PRL secretion from anterior pituitary tissue incubated in vitro. The inhibitory effect of GEP was comparable to that of dopamine in this system. Moreover, haloperidol blocked completely the GEP-mediated suppression of PRL secretion in this preparation. These data suggest agonist properties of both GEP and IPS at D2 dopamine receptors. In light of other evidence that GEP has D2 antagonist effects in vivo, we hypothesize that GEP and perhaps IPS are partial dopamine agonists which may contribute to their antianxiety and/or antidepressant properties. PMID- 2565391 TI - Prolactin-lowering ability of (+/-)-idazoxan may be linked to a central noradrenergic-serotonergic interplay. AB - 2-[2-(1,4-benzodioxanyl)-2-imidazoline] (idazoxan) sometimes lowers basal prolactin levels in the male adult rat, but strongly inhibits hyperprolactinemia in suckling rats. A possible antido paminergic drug effect is not involved, due to its inability to modify prolactin release from superfused pituitary in vitro as well as rat haloperidol hyperprolactinemia in vivo. On the contrary, in the rat idazoxan counteracts hyperprolactinemias due to central presynaptic serotonergic neurotransmission increase (5-hydroxytryptophan, D-fenfluramine and fluoxetine) but not those related to direct agonists at 5-hydroxytryptamine (5 HT) receptor (6-chloro-2-[1-piperazinyl] pyrazine, MK 212; 1-(2,5-dimethoxy-4 iodophenyl)2-aminopropane, DOI). (+/-)-Idazoxan does not modify [3H]-5-HT release from isolated hypothalamic synaptosomes. It displays an affinity for alpha-2 adrenergic autoreceptors about 250-fold more than for alpha-2 heteroreceptors located on 5-HT nerve terminals in the central nervous system (pA2 value 9.99, equivalent to a Kb of 0.1 nM vs. pA2 7.60 equivalent to a Kb of 2.5 nM). The noradrenergic outflow selectively induced by idazoxan in the brain may negatively modulate the 5-HT release from the relevant nerve endings, thus preventing prolactin release due to an activation of presynaptic serotonergic axons induced both physiologically (lactation) and pharmacologically (5-hydroxytryptophan, D fenfluramine and fluoxetine) without influencing hyperprolactinemias related to a direct activation of serotonergic receptors (MK 212 and DOI). Other blocking agents, strong but less selective than (+/-)-idazoxan for noradrenergic brain neurotransmission, do not modify or increase blood prolactin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565392 TI - A nonsteady-state agonist antagonist interaction model using plasma potassium concentrations to quantify the beta-2 selectivity of beta blockers. AB - We studied the competitive interaction of terbutaline and two beta blockers, metoprolol and oxprenolol, with different cardioselectivity for the beta-2 adrenoceptor. Using pharmacokinetic-dynamic modeling in nonsteady-state conditions, of the antagonism by the beta blockers of the terbutaline-induced hypokalemia, the beta blocker beta-2 selectivity was quantitated in the terms of IC50 values representing plasma concentrations resulting in half-receptor occupancy. Six healthy subjects were given an 0.5-mg s.c. dose of terbutaline on three occasions: 1) 1 hr after p.o. administration of a placebo; 2) 1 hr after 150 mg of metoprolol p.o.; and 3) 1 hr after 80 mg of oxprenolol p.o. During 7 hr after terbutaline administration drug concentrations and effects were monitored. Oxprenolol decreased both terbutaline volume of distribution (-69%) and clearance (-63%) and increased its area under plasma concentrations vs. time curve (+157%). Such effects of metoprolol on terbutaline pharmacokinetics were not observed. The dynamic model offered a good description of the observed effects. The apparent IC50 values varied between 42 and 68 ng/ml (mean, 54 ng/ml) for metoprolol and between 3.6 and 4.7 ng/ml (mean, 4.1 ng/ml) for oxprenolol. In view of these results, and comparing them with apparent beta-1 IC50 values as reported in the literature, metoprolol can be considered a relatively beta-1 selective agent. Pharmacokinetic-dynamic modeling of the interaction of beta-2 sympathicomimetics and beta blocking agents after single dosing, seems to be a suitable method for the determination of the relative beta-2 selectivity of the antagonist. PMID- 2565393 TI - Adenosine A1 receptors inhibit adenylate cyclase activity and neurotransmitter release and hyperpolarize pyramidal neurons in rat hippocampus. AB - Adenosine is a neuromodulator with multiple actions upon the physiology and biochemistry of the brain. Although the receptors that inhibit synaptic transmission and regulate cyclic AMP formation have been well characterized in terms of their pharmacological properties, the receptor(s) that mediate the postsynaptic actions of adenosine have not. We have used the adenosine-mediated inhibition of low calcium bursting in the rat hippocampal slice preparation as a measure of a postsynaptic effect of adenosine. Low calcium bursting consists of repetitive spiking evoked by antidromic stimulation of the CA1 pyramidal neurons and can be suppressed by adenosine. In the present study, we compared the effects of a selective adenosine A1 receptor antagonist, 8-cyclopentyltheophylline, on functional responses to adenosine in hippocampal slices. Its apparent affinities for the receptors mediating the following effects were increases in cyclic AMP formation (1700 nM), decreases in cyclic AMP formation after forskolin pretreatment (57 nM), inhibition of excitatory synaptic responses (42 nM), inhibition of repetitive spiking (45 nM), and it was a competitive antagonist for all responses tested. These data demonstrate that inhibition of transmitter release, adenylate cyclase and low calcium bursting in the hippocampus are all mediated via adenosine receptors that have pharmacological properties similar to the A1 receptor and that 8-cyclopentyltheophylline has a high degree of selectivity for this receptor as opposed to the A2 receptor that mediates increases in cyclic AMP formation. PMID- 2565394 TI - Histamine as a central modulator of rat intestinal transit. AB - Histamine (HA) injected i.c.v. to rats inhibited intestinal propulsion in linear relation to the log of the administered doses (in the range from 20-100 micrograms/rat). In the same dose range HA also induced a dose-related analgesic effect (tail-flick test). The dose of HA maximally active by the i.c.v. route (100 micrograms/rat) showed neither of these effects when injected i.v. or i.p. HA-induced intestinal inhibition and analgesia were antagonized competitively by i.c.v. mepyramine (10 micrograms/rat), an H1 receptor antagonist, whereas cimetidine (10 micrograms/rat), an H2 receptor antagonist, had no effect. Repeated i.c.v. injections of HA resulted in tachyphylaxis of both intestinal inhibition and analgesia. Pretreatment with i.c.v. naloxone (20 micrograms/rat) antagonized the antipropulsive effect of HA in a noncompetitive fashion, but did not affect its antinociceptive action. The relevance of the central histaminergic system in the modulation of gastrointestinal motility and its relationship with the opioid system are discussed. PMID- 2565395 TI - Time-dependent enhancement of xylazine-induced, alpha-2 adrenoceptor-mediated vasoconstriction in isolated and perfused canine pulmonary veins. AB - By using the cannula inserting method, vasoconstrictor responses to norepinephrine (a mixed alpha-1 and alpha-2 adrenoceptor agonist), phenylephrine (a selective alpha-1 adrenoceptor agonist), clonidine and xylazine (selective alpha-2 adrenoceptor agonists) were investigated in the isolated and perfused canine pulmonary vein. The segment of vessels was perfused by Krebs-Ringer bicarbonate solution at a constant flow rate at 37 degrees C. Two hours after setting up the perfusion preparation, norepinephrine and phenylephrine induced dose-dependent increases in perfusion pressure. Xylazine and clonidine, however, did not induce any significant responses at this time. Although the vasoconstrictor responses to norepinephrine and phenylephrine did not change statistically during 11 hr after setting up, xylazine-induced responses were perfusion-time-dependently and significantly enhanced 5, 8 and 11 hr after setting up. On the other hand, clonidine induced no significant vascular responses during 11 hr. Xylazine-induced responses were antagonized by DG-5128 (a selective alpha-2 adrenoceptor antagonist), but not by bunazosin (a selective alpha-1 adrenoceptor antagonist). Furthermore, even in the endothelium-removed preparation by treatment with saponin, the enhancement of xylazine-induced responses were similarly observed in a time-dependent manner. These findings suggest that, in the isolated and perfused canine pulmonary vein, xylazine induced alpha-2 adrenoceptor-mediated vasoconstrictor response was time dependently enhanced with the presence and absence of the endothelium. In contrast with xylazine, clonidine did not induce significant vasoconstriction during the experiments. This result suggests that clonidine has little activity at the alpha-2 adrenoceptors which mediate latent xylazine-induced vasoconstriction in the isolated, long-time-perfused canine pulmonary veins. PMID- 2565396 TI - Interaction of a chemically reactive prazosin analog with alpha-1 adrenoceptors of rat tissues. AB - An alkylating analog of prazosin, SZL49 [1-(4-amino-6,7-dimethoxy-2-quinazolinyl) 4-(2-bicyclo[2.2.2]octa-2,5- diene-2-carbonyl)-piperazine], was synthesized and its pharmacological properties examined. SZL49 competed with nanomolar potency at [3H]prazosin binding sites of rat tissues. Pretreatment of membranes with SZL49 (10 nM) followed by washing led to a reduction in [3H]prazosin binding without a change in the Kd of the remaining sites. However, preincubation even at a concentration of 1 microM, led to only a 50% reduction in binding. Higher concentrations of SZL49 in the preincubation mixtures increased the Kd of the remaining sites. Pretreatment of membranes with phenoxybenzamine led to greater than 80% reduction in these sites. Preincubating membranes with SZL49 together with prazosin prevented the loss of binding caused by SZL49 alone. Utilizing different buffers or altering the ratio of absolute amounts of SZL49 and receptors in the preincubations failed to increase the blockade of [3H]prazosin binding sites. SZL49 was injected i.p. into rats and 16 hr later membranes were prepared from tissues and [3H]prazosin saturation experiments were performed. Whereas Kd values in brain and heart were no different from controls, the Kd value of the remaining kidney binding sites was increased approximately 5-fold in test animals in contrast to in vitro experiments. Maximum binding values of heart were reduced significantly by approximately 42%. Maximum binding values of kidney and brain were reduced about 21 and 36%, respectively. In functional studies with isolated rat aorta, pretreatment with SZL49, followed by a 1.5 hr washout, shifted to the right in a dose-dependent manner the dose-response curves for phenylephrine and norepinephrine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565398 TI - The cellular eye lens and crystallins of cubomedusan jellyfish. AB - The ultrastructure and major soluble proteins of the transparent eye lens of two cubomedusan jellyfish, Tripedalia cystophora and Carybdea marsupialis, have been examined. Each species has two complex eyes (one large and one small) on four sensory structures called rhopalia. The lenses consist of closely spaced cells with few organelles. The lens is situated next to the retina, with only an acellular layer separating it from the photoreceptors. SDS-PAGE showed that the large lens of C. marsupialis has only two crystallin polypeptide bands (with molecular masses of approximately 20,000 and 35,000 daltons), while that of T. cystophora has three bands (two with a molecular mass near 20,000 daltons and one with a molecular mass near 35,000 daltons). Interestingly, the small lens of T. cystophora appears to be markedly deficient in or lack the lower molecular weight proteins. The crystallins behaved as monomeric proteins by FPLC and showed no immunological reaction with antisera of the major squid crystallin, chicken delta crystallin or mouse gamma-crystallin in western immunoblots. Very weak reactions were found with antimouse alpha- and beta-crystallin sera. The 35,000 dalton crystallin of T. cystophora was purified and called J1-crystallin. It contained relatively high leucine (13%) and tyrosine (9%) and low methionine (2%). Several tryptic peptides were sequenced. Weak sequence similarities were found with alpha and beta-crystallins, which may account for some of the apparent weak immunological cross-reactivity with these vertebrate crystallins. A polyclonal antiserum made in rabbits from a synthetic peptide of J1-crystallin reacted strongly with J1-crystallin of T. cystophora and C. marsupialis in immunoblots; by contrast, no reaction was obtained with the lower molecular weight crystallins from these jellyfish, with the squid crystallin, or with any crystallins from the frog or human lens. Thus, despite the structural similarities between the cubomedusan, squid and vertebrate lenses, their crystallins appear very different. PMID- 2565397 TI - From distributed sensory processing to discrete motor representations in the diencephalon of the electric fish, Eigenmannia. AB - During their jamming avoidance response (JAR), weakly electric fish of the genus Eigenmannia shift their electric organ discharge (EOD) frequency away from a similar EOD frequency of a neighboring fish. The behavioral rules and neural substrates for stimulus recognition and motor control of the JAR have been extensively studied (see review by Heiligenberg 1986). The diencephalic nucleus electrosensorius (nE) links sensory processing within the torus semicircularis and optic tectum with the mesencephalic prepacemaker nucleus which, in turn, modulates the medullary pacemaker nucleus and hence the EOD frequency. Two separate areas within the nE responsible for JAR-related EOD frequency rises and frequency falls, respectively, were identified by iontophoresis of the excitatory amino acid L-glutamate. Bilateral lesion of the areas causing EOD frequency rises resulted in elimination of JAR-related frequency rises above a baseline frequency obtained in the absence of a jamming stimulus. Similarly, bilateral lesion of the areas causing frequency falls resulted in a loss of JAR-related frequency falls below the baseline frequency. Whether these areas are also responsible for non JAR-related frequency shifts is not known. The strength of response and spatial extent of the areas causing frequency shifts varied among fish and also varied in individual fish, reflecting the strength of JAR-related frequency shifts and the balance of activities in frequency-rise and frequency-fall areas. Local application of bicuculline-methiodide or GABA demonstrated a tonic inhibitory input to each area and suggests a reciprocal inhibitory interaction between the two ipsilateral areas, possibly accounting for much of the individual plasticity. The nE thus is a site for neuronal transformation from distributed, topographically organized processing within the laminated structures of the torus and tectum to discrete cell clusters which control antagonistic motor responses. PMID- 2565399 TI - Synthesis and structure-activity relationship of substituted tetrahydro- and hexahydro-1,2-benzisothiazol-3-one 1,1-dioxides and thiadiazinones: potential anxiolytic agents. AB - Several novel substituted tetrahydro- and hexahydro-1,2-benzisothiazol-3-one 1,1 dioxides and thiadiazinones were prepared and examined in a series of in vitro and in vivo tests to determine their pharmacological profile. Most compounds were orally active in blocking the conditioned avoidance response (CAR) but did not antagonize apomorphine-induced stereotyped behavior. Several compounds demonstrated moderate to high affinity for the 5-HT1A receptor binding site, with compounds 37 and 38 containing 2-pyrimidinylpiperazinyl and [3 (trifluoromethyl)phenyl]piperazinyl moieties and compound 47 containing the 2 pyrazinylpiperazinyl moiety displaying the highest affinity (Ki values of 10, 4, and 9 nM, respectively). Compound 37, 3-[4-[4-(2-pyrimidinyl)-1 piperazinyl]butyl]hexahydro-4, 7-etheno-1H-cyclobut [f]-1,2-benzisothiazol-3(2H) one 1,1-dioxide, buspirone, and ipsapirone showed similarities in their neurochemical and behavioral profiles. They were similar in potency in blocking CAR with AB50 values of 39, 32, and 42 mg/kg, respectively. They also demonstrated high affinity and selectivity for the 5-HT1A receptor site (Ki = 10 nM) and exhibited partial agonist/antagonist activity in the serotonin syndrome test. In addition, compound 37 inhibited apomorphine-induced climbing behavior much more potently (ED50 of 3.4 mg/kg) than stereotyped behavior (ED50 of 32.2 mg/kg) and will be evaluated further. Structure-activity relationships within this series of compounds are discussed. PMID- 2565401 TI - N-(fluoroethyl)(imidazolylphenyl)formamidines. The issue of the active species of mifentidine. AB - Three N-fluoroethyl-substituted (imidazolylphenyl)formamidine derivatives, namely, 2-fluoroethyl (3b), 2,2-difluoroethyl (3c), and 2,2,2-trifluoroethyl (3d), were prepared to test the effect of fluorine substitution on basicity and, then, on H2-antagonist affinity in comparison with the unsubstituted N-ethyl derivative (3a), taken as a model of mifentidine. Imidazolylphenyl isothiocyanate (1), obtained by reaction of 4-(aminophenyl)imidazole with carbon disulfide and ethyl chloroformate, was condensed with the requisite 2-fluoro-substituted ethylamines to give the intermediate thioureas (2b-d). Desulfurization of these thioureas by Raney nickel furnished the desired formamidines (3b-d). Increasing fluorine substitution was found to decrease basicity of the formamidino group substantially (3a, pKa = 8.65; 3b, pKa = 8.12; 3c, pKa = 6.60; 3d, pKa = 6.14), while having a modest effect on the imidazole portion. Affinity at the H2 receptors, evaluated from antagonism of histamine-stimulated chronotropic response on guinea pig atria, increased following fluorine substitution (3a, KB = 177; 3b, KB = 61; 3c, KB = 21; 3d, KB = 7.6). It is concluded that H2-receptor antagonist affinity in the mifentidine series is mostly dependent on the availability of the neutral species. These data support the hypothesis that mifentidine, like cimetidine, acts through the neutral species. PMID- 2565400 TI - Activity of aromatic substituted phenylpiperazines lacking affinity for dopamine binding sites in a preclinical test of antipsychotic efficacy. AB - Generally, antipsychotic agents are dopamine receptor blocking agents that also block conditioned avoidance responding (CAR) in the rat. Recently, however, both (Q-methoxyphenyl)piperazine (OMPP, 1h) and (m-chlorophenyl)piperazine (MCPP, 1o) have been reported to block conditioned avoidance responding in the rat although neither has dopamine receptor blocking properties. The present paper examines the behavioral and biochemical profile of a number of additional substituted phenylpiperazines. None of the phenylpiperazines tested demonstrated high affinity for either dopamine D-1 or D-2 receptor sites, yet many were effective in blocking CAR. The results suggest that the phenylpiperazines may be effective antipsychotic agents without blocking dopamine receptors. Moreover, the active compounds did demonstrate activity in displacing ligand binding to serotonin receptors. Receptor binding profiles were determined for 5-HT-1A and 5-HT-1B binding sites as well as for 5-HT-2 sites. The data from this preclinical test suggest these phenylpiperazines might be effective antipsychotic agents acting via a nondopaminergic mechanism of action. PMID- 2565402 TI - Synthesis and dopaminergic activity of 2-substituted octahydrobenzo[f]quinolines. AB - A series of 2-substituted octahydrobenzo[f]quinolines has been synthesized and assayed for dopamine agonist activity. Only the compounds corresponding to the beta-rotameric conformation of dopamine showed biphasic activity in competition binding studies with the radioligand [3H]spiroperidol. These findings suggest that the congeners possessing the beta-rotamer conformation show receptor-binding characteristics that resemble those of the ergolines more closely than do those of the corresponding alpha-rotamer congeners. PMID- 2565403 TI - Prediction of doxorubicin resistance in vitro in myeloma, lymphoma, and breast cancer by P-glycoprotein staining. AB - Prior studies have shown that the P-glycoprotein is a cell membrane efflux pump that is quantitatively increased in expression in multidrug-resistant tumor cell lines. In this study, fresh tumor tissues from patients with multiple myeloma, malignant lymphoma, or metastatic breast cancer were studied immunohistochemically for P-glycoprotein expression and for in vitro sensitivity to doxorubicin. Twenty-six patients who were either previously untreated or in relapse after chemotherapy had tumor specimens submitted that could be evaluated in both assays. The testing was done independently and blindly in separate laboratories instead of our being provided relevant clinical data on the patients. Tumor cells from 12 of the 26 patients (46%) stained positively for P glycoprotein. Fifteen of the 26 specimens (58%) exhibited drug resistance in vitro. Although only three (21%) of the 14 P-glycoprotein-negative tumors exhibited in vitro resistance to doxorubicin, all 12 fresh tumors that stained positively for P-glycoprotein were resistant to doxorubicin. The difference in frequency of intrinsic doxorubicin resistance between P-glycoprotein-negative and -positive tumors was highly significant (P less than .001). Similar trends were observed in each of the individual tumor categories and were statistically significant in myeloma and breast cancer. Four of the biopsy specimens that stained positively for P-glycoprotein and exhibited doxorubicin resistance were from patients who had not received prior cytotoxic chemotherapy. Similar conclusions were reached when results of drug sensitivity tests were ranked in relation to the median infective dose rather than by criteria based on correlations with clinical drug resistance. Our findings indicate that positive staining for P-glycoprotein associated with multidrug resistance predicts intrinsic cellular resistance of human cancers to doxorubicin. We anticipate that immunohistochemical staining for P-glycoprotein will prove useful in clinical oncology. PMID- 2565404 TI - Increased cytosolic pH in multidrug-resistant human lung tumor cells: effect of verapamil. AB - In a set of four increasingly multidrug-resistant variants of SW-1573 human lung tumor cells, the pHi (i.e., steady-state cytosolic pH) increased up to 0.44 U as a function of the level of doxorubicin resistance. The elevated pHi in the most resistant (2,000-fold) variant dropped to the control level upon addition of verapamil, a known inhibitor of P-glycoprotein activity. These data suggest that, in the absence of xenobiotic substrates, P-glycoprotein activity can affect cellular pHi. This finding may be important for the elucidation of the physiological function of this protein. PMID- 2565405 TI - Drug use in trauma victims. AB - We examined the prevalence and characteristics of drug use in a large sample of fatally and nonfatally injured trauma victims. Routinely collected urine specimens from 452 emergency room patients and 160 persons autopsied at the Medical Examiner's Office (MEO) were analyzed for the presence of marijuana, cocaine, opiates and benzodiazepines using EMIT enzyme immunoassays. Blood alcohol levels were also measured. Tests were positive for at least one drug in 40.3% of the ER and 18.7% of the MEO samples. Marijuana was the most commonly detected drug in both groups. Specimens were more likely to be positive in younger persons and in males, and in victims of assaults and traffic accidents. Alcohol was present in the blood in more than one third of ER and MEO samples. Only 39.8% of ER samples and 52.3% of MEO samples were negative for both alcohol and drugs. PMID- 2565406 TI - Observations on the clinical manifestations and treatment of an experimental infection with Brugia malayi in man. AB - This paper reports clinical observations made during experimental Brugia malayi infection in man. This was induced by 52 infective larvae-carrying mosquitoes, Anopheles sinensis. A monkey, Maccaca mulatta, was inoculated with infective larvae from the mosquitoes infected with nocturnally periodic type of B. malayi from patients. Six months later, the monkey became a microfilaria carrier. Nine days later, the subject was experimentally bitten by mosquitoes infected from biting the monkey, with an estimated delivery of over 200 infective larvae. Seven days after the infection, general (but fluctuating) pruritus developed, followed by lymphadenitis and swelling of the hand and forearm, with a mild pyrexia for nearly 3 months. The eosinophil counts continuously increased. There were four separate attacks of lymphangitis. On day 142, microfilaraemia was first detected. After two more weeks, the subject was treated with a total dose of 15 g diethylcarbamazine (DEC) over 30 days. During this period, 28 small tender subcutaneous nodules appeared on his arms. After treatment, microfilaraemia gradually disappeared and the swelling of his arms improved. Three months later, the eosinophil count of his blood returned to normal but some temporary and scattered small pruritic erythematous patches still persisted over his left forearm. Lymphangitis has not recurred. PMID- 2565407 TI - The number of spermatogonia in various congenital testicular disorders. AB - Various congenital testicular disorders, including monorchism, retractile testis, cryptorchidism and male intersex, were investigated by counting the number of spermatogonia per seminiferous tubule. The results showed that all 7 cases of monorchism had normal numbers of spermatogonia per seminiferous tubule. However, in 29 cases of a retractile testis a normal testis was observed in 13 (44.8 per cent). Therefore testicular dysgenesis is suggested to exist in more than half of cases of the retractile testis. Of 150 cases of cryptorchidism 82 were bilateral and 68 were unilateral. There was no significant difference in the number of spermatogonia per seminiferous tubule between these 2 groups. The higher the testes were located the worse the ratio of spermatogonia per seminiferous tubule. Fewer or absent spermatogonia were observed in 2 patients less than 2 years old. Of 28 contralateral scrotal testes in patients with unilateral cryptorchidism 4 (14.3 per cent) had no spermatogonia per seminiferous tubule and 8 (28.0 per cent) had a decreased number of spermatogonia per seminiferous tubule. The male intersex patients had much damage even in the scrotal testes. From these results it is suggested that these congenital testicular disorders, except monorchism, have similar histological features. Moreover, these conditions are possibly related in etiology to the phenomenon of deficient androgen stimulation. PMID- 2565409 TI - Psychiatry. PMID- 2565408 TI - An in vivo evaluation of alpha adrenergic receptors in canine prostate. AB - The role of alpha 2 adrenergic receptors in the prostate in vivo is unknown. A model was developed to measure canine prostatic urethral pressure in vivo, and to assess the ability of various alpha adrenergic blocking agents to affect prostatic pressure. In this model, an esophogeal pressure catheter is inserted into the prostatic urethra to record prostatic urethral pressure. We investigated the effects of alpha adrenergic agonists and antagonists on prostatic pressure using this model. Dose-response curves were generated for epinephrine, and were then repeated in the presence of either prazosin (alpha 1 antagonist), yohimbine (alpha 2 antagonist) or SK&F-86466 (alpha 2 antagonist). Prazosin was the most potent of the three drugs in competitively blocking epinephrine-induced contraction of the prostate, with an inhibition constant of 0.24 micrograms./kg. calculated from the double reciprocal plot. Clonidine, an alpha 2 adrenergic agonist, caused contraction of the prostate, which was also blocked by prazosin. Furthermore, the specific alpha 2 agonist BHT-920 was totally inactive in our system. These results demonstrate that the increase in urethral pressure caused by alpha-adrenergic agonists can be blocked by alpha adrenergic antagonists. However, the specific alpha 1 antagonist, prazosin, is more potent than alpha 2 antagonists, and is also effective against an alpha 2 agonist, clonidine. This suggests that blockade of alpha 1 receptors may be a more useful strategy for causing relaxation of the prostate than blockade of alpha 2 receptors. PMID- 2565410 TI - [Nursing of patients on respirators. Drugs used during the application of respirators]. PMID- 2565411 TI - Adrenergic control of cAMP generation in rat inner medullary collecting tubule cells. AB - The adrenergic nervous system profoundly alters water excretion by both renal and extrarenal pathways. The effects of catecholamines on cultured rat inner medullary collecting tubule cells were studied. The beta-adrenergic agonist, isoproterenol, increases cAMP from 19.5 +/- 2.3 to 79.4 +/- 14.4 fm/micrograms protein, P less than 0.001. The response to arginine vasopressin (AVP) is also greater in the presence of isoproterenol, but the increment is unchanged when compared to that seen in the absence of AVP. The agonist effect of isoproterenol is blocked by propranolol but not by the specific beta 1 antagonist, atenolol. The effect of alpha-adrenergic stimulation was studied by the use of norepinephrine (NE) in the background of the beta blocker, propranolol. NE decreases AVP-stimulated cAMP generation from 190 +/- 11 to 117 +/- 10 fm/micrograms, P less than 0.001, N = 6. The specific alpha 2 antagonist, yohimbine, but not the alpha 1 antagonist, prazosin, prevents the NE-induced decrease as AVP-stimulated cAMP is restored to 187 +/- 19 fm/micrograms. Similarly the selective alpha 2 agonist, clonidine, significantly inhibits both AVP- and isoproterenol-mediated cAMP generation. To define the site of alpha 2 inhibition in the adenylate cyclase (AC) complex the effect of pertussis toxin (PT) was investigated. After pretreatment with PT (1-1000 ng/ml), AVP-stimulated cAMP was not inhibited by NE. The alpha 1 agonist, phenylephrine, fails to inhibit AC or to increase cytosolic Ca in these cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565412 TI - Immunomodulating therapy of rheumatoid arthritis by high-dose intravenous immunoglobulin. AB - 11 patients with rheumatoid arthritis were treated with intravenous immunoglobulin (IVGG). In 6 patients clinical results were impressive, although lasting responses could be achieved in 3 patients only. This treatment was immunomodulating, since the immunoregulatory T-cell ratio (CD4/CD8) decreased following therapy by reducing CD4-positive cells in-vivo. By use of anti-mu antibodies as a B-cell specific mitogen, IVGG-treatment was seen to suppress early processes of B cell activation. In parallel to these cellular effects, IVGG led to a reduction in the levels of polyethyleneglycol-precipitated circulating immune complexes as measured by lasernephelometry. PMID- 2565413 TI - [Effectiveness of oxygen equipment as a means of the life support system during space flights]. AB - This paper presents pooled data about the factors determining health status and work capacity of aircrew using positive pressure breathing systems at altitudes over 12,000 m. It is indicated that the system compensating positive pressure is primarily responsible for the protective and ergonomic characteristics of altitude equipment. PMID- 2565414 TI - Campylobacter jejuni infection within a laboratory animal production unit. AB - A conventional laboratory animal production unit in which rats, mice, guineapigs and rabbits were bred in one building and cats maintained in a separate, but adjacent area was examined for the presence of intestinal thermophilic Campylobacter spp. Campylobacter jejuni was recovered from 18.84% of 552 animals. The infection rate was highest amongst the cats (51.7%), with rats being the second most commonly infected (23.2%), whereas only 7.7% of guineapigs and a single rabbit (1%) were positive. Campylobacter-like organisms were cultured from 10% of the mice, but these bacteria failed to grow on subsequent subculturing. By using bacterial restriction endonuclease DNA analysis (BRENDA), a single type of C. jejuni was identified from all isolates recovered from the rats, guineapigs and a rabbit, suggesting a common source of infection. In contrast, there were 5 different BRENDA patterns derived from cat isolates. No isolates of C. jejuni were obtained from humans working within the unit or from animal bedding or the immediate environment, although it was suggested that the organism may have entered and spread within the unit from sawdust. PMID- 2565415 TI - Contamination of commercially available quisqualic acid by glutamate-like and aspartate-like substances. AB - Six different batches of the glutamic acid analogue quisqualic acid were analyzed with high-pressure liquid chromatography (HPLC). All batches examined showed contaminant peaks. Different batches had different contaminant peaks and differing amounts of each contaminant. Every batch of quisqualic acid tested demonstrated a contaminant peak which co-eluted with exogenously added glutamic acid. Certain batches possessed a contaminant which co-eluted with aspartic acid. The levels of glutamate-like contamination ranged from 0.08 to 0.60%, and the levels of aspartate-like contamination ranged from undetectable amounts to 0.80%. The amount of combined glutamate- and aspartate-like contamination of each batch of quisqualate correlated very highly with the ability of that batch to interact with non-quisqualate receptors in an autoradiographic binding assay. These non quisqualate receptors are likely N-methyl-D-aspartate (NMDA) receptors. Thus, when high concentrations of quisqualate are used experimentally, contamination is likely to produce spurious effects at non-quisqualate glutamate receptors. Quisqualate itself may be a more specific agonist than assumed previously. PMID- 2565416 TI - The reducing effects of tazifylline on histamine-induced bronchoconstriction in atopic asthmatics. AB - Percentage reductions in the peak flow rates to a standardized number of histamine inhalations were compared after 7 days of treatment with 10 mg bd tazifylline and after 7 days of placebo in a randomized crossover study in 11 male seasonal atopic asthmatics. After tazifylline, mean % reductions in peak flow rates were significantly lower than after placebo (p = 0.03). The effects of tazifylline had disappeared within 7 days of stopping treatment. Using a 50% decrease in bronchial reactivity as the criteria of "protection", 7 of the subjects were "protected" by tazifylline only and none by placebo only (p = 0.03). PMID- 2565417 TI - Prescribed fenoterol and death from asthma in New Zealand, 1981-83: case-control study. AB - A case-control study was conducted to examine the hypothesis that fenoterol by metered dose inhaler (MDI) increases the risk of death in patients with asthma. The case group comprised 117 patients aged 5-45 who died of asthma between August, 1981, and July, 1983. For each case, 4 controls, matched for age and ethnic group, were selected from asthma admissions to hospitals to which the cases themselves would have been admitted, had they survived. The relative risk of asthma death in patients prescribed fenoterol by MDI was 1.55 (95% CI 1.04 2.33, p = 0.03). The possibility of confounding or effect modification by severity was assessed by consideration of subgroups defined by markers of asthma severity. The fenoterol MDI relative risk was 2.21 (95% CI 1.26-3.88, p = 0.01) in patients prescribed three or more categories of asthma drugs, 2.16 (95% CI 1.14-4.11, p = 0.02) in patients with a hospital admission for asthma during the previous 12 months, and 6.45 (95% CI 2.72-15.3, p less than 0.01) in patients prescribed oral corticosteroids at time of death or admission. In the group of patients with the most severe asthma (defined by a hospital admission during the previous year and prescription of oral corticosteroids) the fenoterol MDI relative risk was 13.29 (95% CI 3.45-51.2, p less than 0.01). After adjustment for severity, no other asthma treatment commonly used in New Zealand seemed to be associated with an increased risk of asthma death. Not all sources of bias can be definitely excluded; however, when considered together with other epidemiological and experimental evidence, these findings are consistent with the hypothesis that use of fenoterol by MDI increases the risk of death in severe asthma. PMID- 2565418 TI - Complement system protein C4 and susceptibility to hydralazine-induced systemic lupus erythematosus. AB - 21 patients with systemic lupus erythematosus induced by long-term treatment with hydralazine were investigated to see whether susceptibility to this syndrome was associated with deficiency of the classical pathway complement protein, C4. 16 of 21 (76%) patients had one or more C4 null (ie, non-productive) alleles compared with 35 of 82 normal subjects (43%). This difference was significant. The HLA-DR4 antigen, known to be in linkage disequilibrium with the C4B null allele, was also significantly more frequent in the patients (14 of 21 patients compared with 31 of 81 normal subjects). Susceptibility to hydralazine-induced lupus, as in idiopathic systemic lupus erythematosus, may depend partly upon genetically determined C4 levels. PMID- 2565419 TI - Predisposition to familial osteoarthrosis linked to type II collagen gene. AB - The genetic background of two families, in whom a predisposition to primary osteoarthrosis is inherited as a dominant trait, was investigated. Use of restriction fragment length polymorphisms within and around the type II collagen gene on chromosome 12 revealed a linkage between this cartilage-specific gene and primary osteoarthrosis. PMID- 2565420 TI - Polymerase chain reaction for detection of residual leukaemia. AB - The occasional finding of cells positive for the Philadelphia (Ph) chromosome months or years after bone-marrow transplantation for chronic myeloid leukaemia raises the possibility that the Ph-positive clone may never be eradicated. The polymerase chain reaction with probes able to detect the transcript of the bcr/abl hybrid gene at very low levels was used to study marrow cells from seven patients in continuing haematological and cytogenetic remission 5-7 years after allogeneic bone-marrow transplantation for chronic myeloid leukaemia. No evidence of the leukaemic mRNA was found. Thus, it seems that all leukaemic cells were eradicated in these patients and that they are truly cured. PMID- 2565421 TI - Opioid inhibition of neurally mediated mucus secretion in human bronchi. AB - Capsaicin, which induces release of neuropeptides such as substance P from sensory nerves, stimulated mucus secretion in surgically resected human bronchi in vitro. Pretreatment of the tissue with the opioid antagonist naloxone significantly enhanced secretion, possibly by blocking the inhibitory effect of opiate premedication before surgery. Capsaicin-induced mucus secretion was completely blocked by morphine, and this effect was reversed by naloxone. Thus, sensory nerve stimulation increases mucus secretion in human airways, which might contribute to the mucus hypersecretion seen after inhalation of irritants such as cigarette smoke. Secretion can be completely inhibited by opioid drugs, so they may represent a new therapeutic approach to airway hypersecretion in chronic bronchitis and asthma, in which axon reflex mechanisms have been implicated. PMID- 2565423 TI - Classification of hypertensive disorders of pregnancy. PMID- 2565422 TI - Ligands specific to peripheral benzodiazepine receptors for treatment of porphyrias. AB - Accumulating data indicate that porphyrins are physiologically endogenous ligands to mitochondrial peripheral benzodiazepine receptors. An isoquinoline carboxamide derivative that likewise binds to peripheral benzodiazepine receptors could prove therapeutically useful in porphyrias by displacing porphyrins from these receptors in mitochondria. PMID- 2565424 TI - The new GP contract: will patients suffer? PMID- 2565425 TI - Desmopressin and arterial thrombosis. PMID- 2565426 TI - Investigation of failed low back surgery. PMID- 2565427 TI - Carcinoid tumours and endocrine cell hyperplasia. PMID- 2565428 TI - Nationwide community-based serological survey of HIV-1 and other human retrovirus infections in a central African country. Rwandan HIV Seroprevalence Study Group. AB - In December, 1986, a nationwide serological survey of human immunodeficiency virus type 1 (HIV-1) infection in the general population of Rwanda was done in two parts--one in the rural and the other in the urban population. The sampling method was a modification of the cluster sampling technique developed for monitoring immunisation coverage. Antibodies to HIV-1 (and to HIV-2 and human T cell leukaemia/lymphoma virus type I [HTLV-I]) were detected by immunoenzymatic assays and confirmed by western blot. The number of clusters surveyed was 30 in each setting, urban and rural. HIV-1 seroprevalence was 17.8% (95% confidence interval 14.3-21.2%) in the urban sample (n = 1870) and 1.3% (0.5-2.2%) in the rural sample (n = 742). In the urban sample, females were more frequently HIV-1 seropositive than males (21.0% vs 14.6%). Age-specific peaks of HIV-1 seroprevalence were identified at 0 to 5 years of age (10.1%) and at 26-40 years (30.0%). No differences in seroprevalence were observed in terms of age and sex in the rural sample. None of the sera were seropositive for HIV-2 and HTLV-I seroprevalence was 0.2% in the urban sample and 0.3% in the rural. Nationwide serological surveys could be effective in evaluating the spread of HIV infection and the efficacy of public health interventions against AIDS in developing countries. PMID- 2565430 TI - Working for which patients and at what cost? AB - The Government's white-paper Working for Patients proposes introducing a system in which publicly financed resources for hospital and community health services will be distributed to districts and general practitioners with practice budgets for them to choose between competing providers from both the public and private sectors. The NHS Management Board in 1986 observed that such a system would be costly and impractical and would require careful pilot work in situations where its benefits are likely to outweight its costs. This paper shows that there is no reason for changing that judgement. PMID- 2565429 TI - Interleukins and the immune system 1. PMID- 2565431 TI - Adverse selection: the Achilles heel of the NHS reforms. PMID- 2565432 TI - Aluminium, water chemistry, and Alzheimer's disease. PMID- 2565433 TI - Doubts on "curative" resection of pancreatic cancer. PMID- 2565434 TI - Direct dissolution of gallbladder stones. PMID- 2565435 TI - Juvenile chronic arthritis and coeliac disease. PMID- 2565436 TI - Clinical problems during the fast of Ramadan. PMID- 2565438 TI - Anaphylactoid drug reactions to ciprofloxacin and rifampicin in HIV-infected patients. PMID- 2565437 TI - Gangrenous stomatitis in AIDS. PMID- 2565440 TI - HIV infection--discrimination and criminalisation. PMID- 2565439 TI - Bronchoconstriction, nebulised pentamidine, and mast cells. PMID- 2565441 TI - Parity as prognostic factor for malignant mixed mullerian tumours. PMID- 2565442 TI - Intradermal hepatitis B vaccine. PMID- 2565443 TI - Amyotrophic lateral sclerosis in Sweden 1970-83 and solvent exposure. PMID- 2565444 TI - Trends in mortality from motoneuron disease. PMID- 2565445 TI - Role of routine functional brain imaging in temporal lobe epilepsy. PMID- 2565446 TI - Surgery for temporal lobe epilepsy. PMID- 2565447 TI - Neurological complications after simultaneous immunisation against tick-borne encephalitis and tetanus. PMID- 2565448 TI - Late referral for biliary atresia. PMID- 2565449 TI - Unusual case of haemophilia B. PMID- 2565450 TI - Muscular dystrophy or spinal muscular atrophy? PMID- 2565451 TI - Non-invasive diagnosis of mitochondrial myopathy. PMID- 2565452 TI - Prenatally diagnosed 45,X/46,XY and normal phenotype. PMID- 2565453 TI - Intraperitoneal erythropoietin. PMID- 2565455 TI - Drug monitoring in Spain. PMID- 2565454 TI - Interleukin-2 therapy and blockage of double-lumen catheters. PMID- 2565456 TI - Impotence and nizatidine. PMID- 2565457 TI - Nephrotic syndrome after injections of bovine cartilage and marrow extract. PMID- 2565458 TI - Atropine as possible contaminant of comfrey tea. PMID- 2565459 TI - Dose standardisation of botulinum toxin. PMID- 2565460 TI - Myositis in association with carbimazole therapy. PMID- 2565461 TI - Frequency distribution of serum cholesterol. PMID- 2565462 TI - Insulin secretion and resistance. PMID- 2565463 TI - The insulin pen. PMID- 2565464 TI - Minocycline for dystrophic epidermolysis bullosa. PMID- 2565465 TI - Why is rice water effective for diarrhoea? PMID- 2565466 TI - Successful treatment of falciparum malaria in pregnancy with mefloquine. PMID- 2565467 TI - Escherichia coli O157:H7 from water. PMID- 2565468 TI - Disseminated Fusarium oxysporum infection in patient with heatstroke. PMID- 2565469 TI - Searching for hyperlipidaemia in patients with coronary artery bypass grafts. PMID- 2565470 TI - Cholera and contagion. PMID- 2565471 TI - Court of appeal endorses medical decision to allow baby to die. PMID- 2565472 TI - Cases/controls, breast cancer, and the pill. PMID- 2565473 TI - Con-fusion. PMID- 2565475 TI - Interleukins and the immune system 2. PMID- 2565474 TI - Duration of impaired consciousness in relation to side of lesion after severe head injury. NIH Traumatic Coma Data Bank Research Group. AB - The relation between hemispheric lateralisation of intracerebral lesion and duration of impaired consciousness was investigated in 43 survivors of severe closed head injury. Duration of impaired consciousness, defined as the time from injury until the obeying of commands, was significantly longer in patients with left hemisphere than right hemisphere lesions. However, the side of intracerebral lesion had no effect on duration of impaired consciousness defined as time from injury until localisation to pain. Use of verbal methods to assess level of consciousness may be responsible for the observation that lesions of the language dominant hemisphere produce greater disturbance of consciousness than similar insults to the right hemisphere. PMID- 2565476 TI - Relative weight and major ischaemic heart disease events in hypertensive men. AB - Some studies have suggested that lean hypertensive men may be at greater risk of major ischaemic heart disease (IHD) events than obese hypertensive men. The issue was examined on data from the British Regional Heart Study for 7735 middle-aged men followed up for an average of 7.5 years; during this time 443 men experienced a major IHD event. Hypertension was defined as systolic blood pressure of 160 mm Hg or above, diastolic blood pressure of 95 mm Hg or above, or receiving treatment for hypertension. For both hypertensive and normotensive men the rate of major IHD events, standardised for age and cigarette smoking, rose with increasing body mass index (BMI). The relative odds associated with a 5 kg/m2 difference in BMI (ie, a 15 kg difference in weight in men of average height [1.73 m]) were 1.30 (p = 0.02) and 1.43 (p = 0.0004) for hypertensive and normotensive men, respectively. A review of eleven prospective studies, including the British Regional Heart Study, presented in standard form for comparative purposes, suggests that lean hypertensive men are not at higher risk of major IHD events than overweight/obese hypertensive men. There seems to be no justification for the suggestion that a policy of weight reduction to lower blood pressure might be inappropriate. PMID- 2565477 TI - Good clinical practice in clinical research. PMID- 2565478 TI - Community outbreak of Legionnaires' disease in Barcelona. PMID- 2565479 TI - Anaesthetic agents and the ozone layer. PMID- 2565480 TI - Indoor air pollution in developing countries and acute respiratory infection in children. PMID- 2565481 TI - Increased risk of breast cancer after low-dose irradiation. PMID- 2565482 TI - Reduced platelet aggregation in long-distance runners. PMID- 2565483 TI - Rapid test for malaria. PMID- 2565484 TI - Novel KI-ras codon 61 mutation in infiltrating leucocytes of oral squamous cell carcinoma. PMID- 2565485 TI - Diagnosis of Niemann-Pick disease type C on chorionic villus cells. PMID- 2565486 TI - Treatment of membranous nephropathy with prednisolone and chlorambucil. PMID- 2565487 TI - Indocyanine-green clearance and liver transplantation. PMID- 2565488 TI - Nausea and vomiting after general anaesthesia. PMID- 2565489 TI - Haemolysis after intravenous streptokinase. PMID- 2565490 TI - Hypokalaemia in patients with acute myeloid leukaemia after treatment with fluconazole. PMID- 2565491 TI - Severe anorexia and possible psychosis or hypomania after trazodone-tryptophan treatment of aggression. PMID- 2565492 TI - A new treatment for endometriosis. PMID- 2565493 TI - Campylobacter pylori, non-ulcer dyspepsia, and race in Malaysia. PMID- 2565494 TI - Rising mortality from motoneuron disease. PMID- 2565495 TI - Does adult T-cell leukaemia originate from intestinal mucosa. PMID- 2565496 TI - Long-term evaluation of mechanical heart valves. PMID- 2565497 TI - Fetal brain tissue and Parkinson's disease. PMID- 2565498 TI - Pamidronate to reduce bone pain in normocalcaemic patient with disseminated prostatic carcinoma. PMID- 2565499 TI - Electric shock treatment for snake bite. PMID- 2565500 TI - Erythromycin resistance in group A streptococci. PMID- 2565501 TI - Hypoxic-reperfusion injury in inflamed joints. PMID- 2565502 TI - Soy-based infant milk formulas and passive intestinal permeability. PMID- 2565503 TI - Levels of aluminium in infant formulae. PMID- 2565505 TI - Scandal in Southwark. PMID- 2565504 TI - Adoptive immunotherapy. PMID- 2565506 TI - In-vitro fertilisation and sex ratio. PMID- 2565507 TI - Influence of zidovudine on progression to AIDS in cohort studies. PMID- 2565508 TI - Jaundice in HIV positive haemophiliac. PMID- 2565509 TI - Reduced CD4+ T cells and candidiasis in absence of HIV infection. PMID- 2565510 TI - Removal and replacement of Tenckhoff catheter at single operation. PMID- 2565511 TI - VP-1 antigen in chronic postviral fatigue syndrome. PMID- 2565512 TI - Erythropoietin in acute renal failure. PMID- 2565513 TI - Lipid-lowering drugs in treatment of hyperlipidaemia associated with nephrotic syndrome. PMID- 2565514 TI - AIDS and HIV: a myth? PMID- 2565515 TI - Oral contraceptive use and breast cancer risk in young women. UK National Case Control Study Group. AB - All women diagnosed with breast cancer before age 36 in eleven geographical areas in Britain were included in a case-control study designed to investigate the relation between oral contraceptive (OC) use and breast cancer risk. 755 cases each with a matched control were interviewed in their homes by trained interviewers. General practitioner notes and family planning clinic records were abstracted and combined with the interview data. There was a highly significant trend in risk of breast cancer with total duration of OC use with relative risks of 1.43 (95% confidence interval 0.97-2.12) for 49-96 months use, and 1.74 (95% confidence interval 1.15-2.62) for 97 or more months use. The relative risks were similar for use before and after first full-term pregnancy. There is some evidence that OCs containing less than 50 micrograms oestrogen have a lower risk associated with their use than higher oestrogen dose OCs and that there may be some protective effect of use of progestagen-only pills. These apparent differences between OC types are only marginally significant statistically and need further investigation. Extensive investigations of possible bias using data abstracted from general practitioner notes for both interviewed and non interviewed cases and controls demonstrate that biases cannot explain these results although some relative risks may be slightly exaggerated. There is, however, no support for these findings in national breast cancer registration rates which are not increasing. PMID- 2565516 TI - Association between biological properties of human immunodeficiency virus variants and risk for AIDS and AIDS mortality. AB - 49 individuals seropositive for human immunodeficiency virus (HIV) antibody were studied longitudinally for the relation between in-vitro properties of their sequential HIV isolates and clinical course before and after the development of AIDS. They were classified into three groups according to the syncytium-inducing capacity, replication rate, and host range of their HIV isolates. The most rapid progression to AIDS (median 15 months) and the lowest survival rate following AIDS diagnosis (median survival 12.5 months) were observed in individuals with high-replicating, syncytium-inducing HIV isolates, followed by individuals with high-replicating, non-syncytium-inducing isolates. In contrast, most individuals with low-replicating, non-syncytium-inducing HIV isolates remained symptom-free during the study period (median follow-up until AIDS diagnosis greater than 42 months), and the few individuals from this group in whom AIDS developed were still alive at the end of the study period (median survival greater than 34 months). In addition, AIDS patients from the three groups differed with respect to their symptoms. Zidovudine treatment in the symptom-free period seemed to delay the onset of AIDS in all risk groups, although stabilisation of CD4+ cell numbers was observed only in individuals with non-syncytium-inducing HIV variants. PMID- 2565517 TI - Beneficial effect of brief pre-transfusion incubation of platelets at 37 degrees C. AB - The effects of brief warming of stored platelet concentrates were assessed in 15 children undergoing transfusion for stable thrombocytopenia due to chemotherapy (n = 13) or aplastic anaemia. Half of a pool of platelet concentrates stored at 22 degrees C was incubated at 37 degrees C for 1 hour and the other half at room temperature. Each patient received one bag of warmed and one of unwarmed cells transfused in random order 2 h apart. Platelet warming improved transfusion efficacy, as assessed on the basis of corrected platelet count increments (CCIs) and platelet morphology. Compared with unwarmed bags, warmed bags had a higher morphology score (p = 0.0001) and a higher CCI (adjusted for the transfusion order) at 1 h (n = 11; p = 0.014) and at 2 h (n = 15, p = 0.006) post transfusion. Thus, with platelets stored at room temperature bags warmed before transfusion to 37 degrees C for 1 h provide a larger number of circulating platelets after transfusion than do unwarmed bags. PMID- 2565518 TI - Plasma endothelin levels in patients with uraemia. AB - Plasma immunoreactive endothelin concentrations were measured in patients with uraemia and in non-uraemic controls. Concentrations were beneath the detection limit of the assay in most patients without uraemia but were readily detectable in those undergoing haemodialysis. Plasma endothelin concentrations in patients with uraemia who were not undergoing haemodialysis were lower than in those on maintenance dialysis. PMID- 2565519 TI - Nutrition and the metabolic response to injury. PMID- 2565520 TI - Ascariasis. PMID- 2565521 TI - Cosines and sinus arrhythmia. PMID- 2565522 TI - Inhaled steroids and recurrent wheeze after bronchiolitis. PMID- 2565523 TI - The management of acute poisoning due to beta-adrenoceptor antagonists. AB - Although many cases of beta-adrenoceptor antagonist (beta-blocker) poisoning are uneventful, a proportion develop serious and sometimes fatal cardiovascular system depression with severe hypotension. As beta-adrenergic tone is not essential for cardiovascular function in health, there is no physiological reason why total beta-adrenoceptor blockade should have serious consequences in the resting individual. The toxic actions of beta-blockers appear to be related to properties such as membrane depressant activity and possibly due to actions on beta-adrenoceptors distinct from those in the cardiovascular system. Most reports of serious adverse effects following overdosage concern beta-blockers with significant membrane depressant activity, and in particular propranolol and oxprenolol, with which progressive heart block and bradycardia are features. Sotalol toxicity, with its unique electrophysiological action, is a special case. Animal experiments confirm that beta-blockers with membrane depressant activity are more toxic than the newer more selective ones, such as atenolol and nadolol. However, experimental models also reveal that artificial ventilation markedly reduces the toxicity of all beta-blockers tested, suggesting a respiratory depressant action with very high doses. Treatment of serious overdosage in man should include maintenance of adequate ventilation. High dose intravenous glucagon is recommended, because its inotropic action depends on direct stimulation of adenylate cyclase. beta-Agonists such as isoprenaline (isoproterenol) or prenalterol may be effective, but the nature of agonist competitive antagonist interactions may necessitate the use of unrealistically large doses to overcome very high tissue beta-blocker concentrations. PMID- 2565524 TI - Astemizole--another non-sedating antihistamine. PMID- 2565525 TI - [New data on the fauna of blood-sucking Culicidae mosquitoes in Ziiama forest (Forest Guinea)]. AB - A total of 885 specimens of blood-sucking mosquitos from Zyama Forest (the Republic of Guinea) were identified as belonging to 38 species of 8 genera. Three species were recorded there for the first time whereas 27 species, 2 subgenera and 2 genera were new for the studied region of the country (Forest Guinea). Four specious combinations of the mosquitos were distinguished as anthropophilic, sylvatic, meadow and marsh ones. PMID- 2565526 TI - [Morphofunctional changes in the ovarioles of blood-sucking mosquitoes (Diptera, Culicidae) during oogenesis. I. Normal oogenesis]. AB - Contemporary level of knowledge of morphological specificities of the mosquitos' ovariole development during normal oogenesis is analysed basing on the authors' proper research and literature data. Disputable problems of morphofunctional role of certain ovariole structures are discussed. The paper is illustrated by photos and pictures of ovarioles from the moment of germarium formation to the beginning of vitellogenesis. PMID- 2565527 TI - [Mixed infections in the pathology of blood-sucking larvae pathology. 2. Entomopathogenic properties of bacterioviral complexes]. AB - Mixed infection of the mosquitoes' larvae of the first age group by densonucleasis virus and entomopathogenic strains of Pseudomonas fluorescens and Serratia marcescens enhanced viral infection in the presence of toxicosis induced by exogenic entomotoxic bacterial metabolites. Possibility of interaction between bacterial cells and the mosquito densonucleasis virus, producing an adverse effect on the duration of the disease, was demonstrated. Duration of the disease provoked by bacterioviral infection of the larvae was of a specific character: nontypical degeneration and untimely replacement of intestinal epithelium at the fourth larval stage were observed along with acute viral lesions of all the tissues. PMID- 2565528 TI - [The Nobel Prize for Medicine for the development of a therapeutic principle]. PMID- 2565529 TI - [Pharmacologic prevention of allergic complications caused by local anesthesia in odontostomatology]. AB - The immunological responses of IgE mediated Type I and of cell-mediated Type IV that intervene in the biphasic mechanism of allergic phenomena, represent the site of action of the pharmacological therapeutic aids usable in the prophylaxis of allergic complications of local anaesthetics. Having examined the pharmacodynamics, the possible clinical use of these drugs is discussed. PMID- 2565530 TI - The C proteins of HeLa 40S nuclear ribonucleoprotein particles exist as anisotropic tetramers of (C1)3 C2. AB - The C proteins (C1 and C2) of HeLa 40S heterogeneous nuclear ribonucleoprotein particles copurify under native conditions as a stable complex with a fixed molar protein ratio (S.F. Barnett, W.M. LeStourgeon, and D.L. Friedman, J. Biochem. Biophys. Methods 16:87-97, 1988). Gel filtration chromatography and velocity sedimentation analyses of these complexes revealed a large Stokes radius (6.2 nm) and a sedimentation coefficient of 5.8S. On the basis of these values and a partial specific volume of 0.70 cm3/g based on the amino acid composition, the molecular weight of the complex was calculated to be 135,500. This corresponds well to 129,056, the sequence-determined molecular weight of a (C1)3C2 tetramer. Reversible chemical cross-linking with dithiobis(succinimidyl propionate) and analysis of cross-linked and cleaved complexes in sodium dodecyl sulfate polyacrylamide gel electrophoresis confirmed that the C proteins exist as tetramers, most or all of which are composed of (C1)3C2. The tetramer is stable in a wide range of NaCl concentrations (0.09 to 2.0 M) and is not dissociated by 0.5% sodium deoxycholate. This stability is not the result of disulfide bonds or interactions with divalent cations. The hydrodynamic properties of highly purified C-protein tetramers are the same for C-protein complexes released from intact particles with RNase or high salt. These findings support previous studies indicating that the core particle protein stoichiometry of 40S heterogeneous nuclear ribonucleoproteins is N(3A1-3A2-1B1-1B2-3C1-1C2), where N = 3 to 4, and demonstrate that the C-protein tetramer is a fundamental structural element in these RNA-packaging complexes. The presence of at least three tetramers per 40S monoparticle, together with the highly anisotropic nature of the tetramer, suggesting that one-third of the 700-nucleotide pre-mRNA moiety packaged in monoparticles is associated through a sequence-independent mechanism with the C protein. PMID- 2565531 TI - Purification of a cellular replication factor, RF-C, that is required for coordinated synthesis of leading and lagging strands during simian virus 40 DNA replication in vitro. AB - Cell extracts (S100) derived from human 293 cells were separated into five fractions by phosphocellulose chromatography and monitored for their ability to support simian virus 40 (SV40) DNA replication in vitro in the presence of purified SV40 T antigen. Three fractions, designated I, IIA, and IIC, were essential. Fraction IIC contained the known replication factors topoisomerases I and II, but in addition contained a novel replication factor called RF-C. The RF C activity, assayed in the presence of I, IIA, and excess amounts of purified topoisomerases, was detected in both cytosol and nuclear fractions, but was more abundant in the latter fraction. RF-C was purified from the 293 cell nuclear fraction to near homogeneity by conventional column chromatography. The reconstituted reaction mix containing purified RF-C could replicate SV40 origin containing plasmid DNA more efficiently than could the S100 extract, and the products were predominantly completely replicated, monomer molecules. Interestingly, in the absence of RF-C, early replicative intermediates accumulated and subsequent elongation was aberrant. Hybridization studies with strand-specific, single-stranded M13-SV40 DNAs showed that in the absence of RF C, abnormal DNA synthesis occurred preferentially on the lagging strand, and leading-strand replication was inefficient. These products closely resembled those previously observed for SV40 DNA replication in vitro in the absence of proliferating-cell nuclear antigen. These results suggest that an elongation complex containing RF-C and proliferating-cell nuclear antigen is assembled after formation of the first nascent strands at the replication origin. Subsequent synthesis of leading and lagging strands at a eucaryotic DNA replication fork can be distinguished by different requirements for multiple replication components, but we suggest that even though the two polymerases function asymmetrically, they normally progress coordinately. PMID- 2565532 TI - The poly(A)-poly(A)-binding protein complex is a major determinant of mRNA stability in vitro. AB - Using an in vitro mRNA decay system, we investigated how poly(A) and its associated poly(A)-binding protein (PABP) affect mRNA stability. Cell extracts used in the decay reactions were depleted of functional PABP either by adding excess poly(A) competitor or by passing the extracts over a poly(A)-Sepharose column. Polyadenylated mRNAs for beta-globin, chloramphenicol acetyltransferase, and simian virus 40 virion proteins were degraded 3 to 10 times faster in reactions lacking PABP than in those containing excess PABP. The addition of purified Saccharomyces cerevisiae or human cytoplasmic PABP to PABP-depleted reactions stabilized the polyadenylated mRNAs. In contrast, the decay rates of nonpolyadenylated mRNAs were unaffected by PABP, indicating that both the poly(A) and its binding protein were required for maintaining mRNA stability. A nonspecific single-stranded binding protein from Escherichia coli did not restore stability to polyadenylated mRNA, and the stabilizing effect of PABP was inhibited by anti-PABP antibody. The poly(A) tract was the first mRNA segment to be degraded in PABP-depleted reactions, confirming that the poly(A)-PABP complex was protecting the 3' region from nucleolytic attack. These results indicate that an important function of poly(A), in conjunction with its binding protein, is to protect polyadenylated mRNAs from indiscriminate destruction by cellular nucleases. A model is proposed to explain how the stability of an mRNA could be affected by the stability of its poly(A)-PABP complex. PMID- 2565535 TI - Structural differences in the renin gene of Dahl salt-sensitive and salt resistant rats. AB - Genomic libraries in lambda EMBL4 phage were constructed from both inbred Dahl salt-hypertension-sensitive (S) and inbred Dahl salt-hypertension-resistant (R) rats. Overlapping clones containing the renin genes were isolated from these libraries by screening with a renin cDNA probe. Clones were characterized by a combination of restriction mapping and Southern blot analysis. The results showed that the S-rat renin gene is remarkably different from the R-rat renin gene. The major differences are 1) a 1.2-kilobase (kb) insertion in the first intron of the S-gene which accounts for most of the restriction fragment length polymorphisms found in the renin genes between S and R strains, such as those generated with Bg/II [2.7 kb (S)/1.5 kb (R)], EcoRI [6.4 kb (S)/5.2 kb (R)], and HindIII [9.6 kb (S)/8.4 kb (R)]; 2) an additional HindIII site located at the 3' end of the R gene which accounts for another HindIII restriction fragment length polymorphisms [25 kb (S)/22 kb, 3.4 kb (R)]; 3) two SmaI sites at the 5' flanking region of the first exon of the S-gene, whereas there is only one SmaI site in the corresponding region of the R-gene; and 4) three AvaI sites in the first intron of the S-gene in contrast to two AvaI sites in the same region of the R-gene These differences in the renin genes of Dahl rats might affect renin gene expression, which could account for the known strain differences in plasma and tissue renin activities. These structural studies provide a basis for genetic investigation into the relationship of the renin gene to blood pressure in Dahl rats. PMID- 2565533 TI - The regulated production of mu m and mu s mRNA is dependent on the relative efficiencies of mu s poly(A) site usage and the c mu 4-to-M1 splice. AB - The relative abundance of the mRNAs encoding the membrane (mu m) and secreted (mu s) forms of immunoglobulin mu heavy chain is regulated during B-cell maturation by a change in the mode of RNA processing. Current models to explain this regulation involve either competition between cleavage-polyadenylation at the proximal (mu s) poly(A) site and cleavage-polyadenylation at the distal (mu m) poly(A) site [poly(A) site model] or competition between cleavage-polyadenylation at the mu s poly(A) site and splicing of the C mu 4 and M1 exons, which eliminates the mu s site (mu s site-splice model). To test certain predictions of these models and to determine whether there is a unique structural feature of the mu s poly(A) site that is essential for regulation, we constructed modified mu genes in which the mu s or mu m poly(A) site was replaced by other poly(A) sites and then studied the transient expression of these genes in cells representative of both early- and late-stage lymphocytes. Substitutions at the mu s site dramatically altered the relative usage of this site and caused corresponding reciprocal changes in the usage of the mu m site. Despite these changes, use of the proximal site was still usually higher in plasmacytomas than in pre-B cells, indicating that regulation does not depend on a unique feature of the mu s poly(A) site. Replacement of the distal (mu m) site had no detectable effect on the usage of the mu s site in either plasmacytomas or pre-B cells. These findings are inconsistent with the poly(A) site model. In addition, we noted that in a wide variety of organisms, the sequence at the 5' splice junction of the C mu 4 to-M1 intron is significantly different from the consensus 5' splice junction sequence and is therefore suboptimal with respect to its complementary base pairing with U1 small nuclear RNA. When we mutated this suboptimal sequence into the consensus sequence, the mu mRNA production in plasmacytoma cells was shifted from predominantly mu s to exclusively mu m. This result unequivocally demonstrated that splicing of the C mu 4-to-M1 exon is in competition with usage of the mu s poly(A) site. A key feature of this regulatory phenomenon appears to be the appropriately balanced efficiencies of these two processing reactions. Consistent with predictions of the mu s site-splice model, B cells were found to contain mu m precursor RNA that had undergone the C mu 4-to-M1 splice but had not yet been polyadenylated at the mu m site. PMID- 2565534 TI - Retrovirus-mediated transfer and expression of the interleukin-3 gene in mouse hematopoietic cells result in a myeloproliferative disorder. AB - A high-titer, recombinant retroviral vector produced in psi 2 packaging cells has been used to introduce the murine interleukin-3 (IL-3) gene into mouse hematopoietic cells. Integration and expression of the IL-3 gene was observed in spleen foci from which could be derived factor-independent, continuously proliferating cell lines. Irradiated or genetically anemic W/Wv recipients of infected hematopoietic cells developed a myeloproliferative syndrome characterized by a marked elevation in leukocyte count, bone marrow hyperplasia, and enlargement of the liver and spleen. The syndrome reflected proliferation of one or more stem cell clones, the progeny of which were capable of repopulating secondary recipients. One animal developed the syndrome primarily by a paracrine mechanism. Endogenous IL-3 production caused amplification of hematopoietic cells but did not appear to alter the maturational or self-renewal potential of these cells. PMID- 2565536 TI - [Antihypertensive agents: adrenergic beta receptor agonists]. PMID- 2565537 TI - [Kidney tubule dysfunction caused by acetic acid]. AB - Two patients admitted after ingestion of 80% acetic acid are described. Only the first patient developed haemolysis, slight intravascular coagulation and oliguric kidney insufficiency. They were treated with a nasogastric tube and total parenteral feeding. During the first week after admission urinary excretion of beta 2-microglobulin, alanine-aminopeptidase and N-acetyl-glucosaminidase was significantly increased. The patients remained haemodynamically stable and did not develop fever. The above-mentioned elevated excretions returned to normal levels. Both patients showed similar patterns of tubular proteinuria. The observations in the second patient suggest a direct toxic effect of acetic acid on the proximal tubule of the kidney. PMID- 2565538 TI - [Disorders in male fertility during treatment with sulfasalazine]. PMID- 2565539 TI - Chronic renal tubular effects in relation to urine cadmium levels. AB - In this study the urine activities of two brush border membrane enzymes, alanine aminopeptidase (AAP) and gamma-glutamyltranspeptidase (GGT), and the lysosomal enzyme N-acetyl-beta-D-glucosaminidase (NAG) were measured in men exposed to cadmium to investigate chronic renal toxicity. The subjects consist of a group with urine cadmium levels less than 2.0 micrograms/l and a group with higher cadmium levels (urine cadmium greater than or equal to 2.0 micrograms/l) with past or present occupational exposure to cadmium. The mean NAG value corrected for creatinine in the higher cadmium group (2.95 U/g creatinine) is significantly different from the mean low cadmium group value (0.92 U/g creatinine, p = 0.0083). Likewise, AAP in the higher group (13.83 U/g creatinine) is significantly different from that of the low group (6.58 U/g creatinine; p = 0.0018). NAG and AAP also give significant correlations with cadmium levels in urine (NAG: r = 0.51; p = 0.0001; AAP: r = 0.56; p = 0.0001). There is no similar statistically significant difference in GGT means in the two groups. In contrast to cadmium correlations, blood lead does not correlate with NAG, AAP, or GGT. A dose-response relationship was found between NAG and cadmium and between AAP and cadmium. The analysis of this relationship gives estimates of a 10% chance of observing an elevated NAG value at a cadmium level of 6.3 micrograms/g creatinine (8.0 micrograms/l) and a 10% chance of observing an elevated AAP at a cadmium level of 5.0 micrograms/g creatinine (3.4 micrograms/l). These data indicate elevations of NAG and AAP at urine cadmium levels below the level of 10 micrograms/g creatinine recommended as an upper limit by the 1980 World Health Organization Study Group. Since elevated levels of NAG and AAP in cadmium-exposed workers may reflect chronic renal tubular nephrotoxicity, these findings indicate that cadmium levels below 10 micrograms/g creatinine may be accompanied by subclinical tubular dysfunction, and that WHO guidelines should be interpreted cautiously, particularly with reference to workers who are no longer exposed and may have had higher cadmium body burdens in the past. PMID- 2565540 TI - Increased seizure susceptibility induced by guanidinoethane sulfonate in E1 mice and its relation to glutamatergic neurons. AB - Convulsions and brain levels of amino acids and 5-hydroxytryptamine (5-HT) in E1 mice were examined after oral administration of a 1% guanidinoethane sulfonate (GES) solution. The incidence of convulsions increased 3 days after starting GES administration, and this effect continued throughout the 6 months of drug administration. Glutamate levels were increased in the cerebrum, and glutamine levels were increased in the cerebellum three days after starting GES administration. Brain 5-HT levels were not changed at that time. These results suggest that increased seizure susceptibility induced by GES in E1 mice is related to glutamatergic neurons. PMID- 2565541 TI - Study of amino acid formation during palmitate oxidation in rat brain mitochondria. AB - The interrelation of palmitate oxidation with amino acid formation in rat brain mitochondria has been investigated in purified mitochondria of nonsynaptic origin by measuring the formation of aspartate, alpha-ketoglutarate, and glutamate during palmitate oxidation, and also by assaying 14C-products of [1-14C]palmitate oxidation. Oxidation of palmitate (or [1-14C]palmitate) resulted in the formation of aspartate (or 14C-aspartate), and the oxidation was inhibited by aminooxyacetate (an inhibitor of transaminase). Palmitate oxidation also resulted in alpha-ketoglutarate formation, which was sensitive to the effect of aminooxyacetate. Addition of NH4Cl was found to increase 14C-products and formation of alpha-ketoglutarate, whereas glutamate formation was not increased unless the rate of palmitate oxidation was reduced by 50% by aminooxyacetate or alpha-ketoglutarate was added exogenously. Exogenous alpha-ketoglutarate was found to decrease 14C-products, but not aspartate formation. These results indicated that palmitate oxidation was closely related to aspartate formation via aspartate aminotransferase. During palmitate oxidation without aminooxyacetate or added alpha-ketoglutarate, however, alpha-ketoglutarate was not available for glutamate formation via glutamate dehydrogenase. We discuss the possibility that this was because (a) oxidative decarboxylation of alpha-ketoglutarate to form succinyl-CoA was favored over glutamate formation for the competition for alpha ketoglutarate in the same pool, and (b) the pool of alpha-ketoglutarate produced in the aspartate aminotransferase reaction did not serve as substrate for glutamate formation. PMID- 2565542 TI - Isolation of metabolically distinct synaptosomes on Percoll gradients. AB - Synaptosomes were prepared from whole rat brain by six different methods based on gradients of sucrose, Ficoll or Percoll. In these, the synthesis and calcium specific release of amino acids were assessed by two different procedures. Preparations based on sucrose showed the least calcium-specific release, followed by Ficoll-derived synaptosomes. As previously described, Percoll gave two separate populations of synaptosomes, both very active in terms of release of aspartate, glutamate, and GABA. The data involving release and synthesis were not identical, but did agree in the following: in low-density synaptosomes, haloperidol blocked both the release and synthesis of glutamate, but was without effect in the heavier population. 2-chloroadenosine and 2-oxoglutarate affected both release and synthesis only in the high-density population. Dopamine blocked aspartate release and synthesis only in the high-density population. These results suggest that haloperidol interferes with glutamate release and synthesis via a mechanism which may not involve adenosine, serotonin, or dopamine. PMID- 2565543 TI - Microspectrofluorometric evaluation of single- and double-stranded DNA in short term cultured human glioma cells. AB - Seven cerebral gliomas in short-term culture were studied by microspectrofluorometry and acridine orange staining to assess their nuclear content of single- and double-stranded DNA. Benign gliomas showed a diploid DNA pattern, whereas malignant gliomas revealed a higher frequency of DNA aneuploidy and hyperploidy. The content of single-stranded DNA remained relatively low in benign gliomas; however, that in malignant gliomas varied widely. After ACNU treatment, the double-stranded DNA histograms showed S-phase-specific accumulation in all cases but 1 with increasing concentrations of ACNU. The single-stranded DNA content decreased considerably in 2 cases, which responded well to chemotherapy and showed clinical amelioration. PMID- 2565544 TI - Effect of magnesium valproate on amygdala-kindled seizures in the rat: comparison with sodium valproate. AB - The anticonvulsant activity of a salt of valproic acid (VA), magnesium valproate (MgV), was assessed against amygdala-kindled seizures in rats. The anti-epileptic power of MgV was compared with that of sodium valproate (NaV). Kindling was obtained by delivering daily to one of the amygdala a 2 s train of monophasic square-wave pulses (1 ms, 60 c.p.s., 100-130 microA) via chronically implanted electrodes. Magnesium valproate and NaV were tested once kindling was stabilized and the post-kindling threshold for generalized convulsions was determined. The drugs were administered intraperitoneally in doses ranging from 25 to 200 mg/kg. The injection/test interval was 30 min. Each animal received a single dose every 24 h. Magnesium valproate exhibited an anticonvulsant activity qualitatively and quantitatively similar to that of NaV. Statistically significant differences were not found between the two drugs with respect to the reduction of seizure severity and afterdischarge (AD) duration. The calculated ED50's were 94.58 and 97.41 mg/kg for the suppression of generalized seizures, 176.96 and 129.26 mg/kg for the suppression of partial seizures, 275.96 and 224.13 mg/kg for the suppression of the local AD in the MgV and NaV treated groups, respectively. PMID- 2565545 TI - Central nervous system lymphomas and immunodeficiency. AB - Primary lymphomas of the central nervous system (CNS) may sometimes be associated with some immunological abnormalities, including renal or cardiac transplants, some congenital and acquired immunodeficiencies, immunoinflammatory diseases and immunosuppressive treatments. A relatively high incidence of cerebral lymphomas has been particularly noticed in renal or cardiac transplantation patients and in those with acquired immune deficiency syndrome (AIDS), two conditions which are today observed with increasing frequency. The different congenital and acquired immunodeficiencies associated with cerebral lymphomas and the pathogenetic connections between the two conditions are discussed from a large review of the literature. PMID- 2565547 TI - Relationship between oedema, blood pressure, and blood flow following local brain injury. AB - Vasogenic brain oedema is a concomitant of a wide variety of central nervous system lesions that have in common a disturbance of the blood-brain barrier permeability. Although it is well established that the formation and spread of this extracellular type of oedema is influenced directly by variations of systemic arterial pressure, the consequences of these events on local cerebral blood flow are not known. In the current experiments, vasogenic brain oedema was produced in rats by cold-injury lesions, and local cerebral blood flow measurements were made using a technique of 14C-antipyrine quantitative autoradiography under pharmacologically controlled conditions of hypertension, hypotension, and normotension. Evidence is presented that the formation and spread of vasogenic brain oedema is accompanied by the development of a congruent zone of decreased local cerebral blood flow, and that the magnitude of this field of oedema/ischaemia is a direct function of systemic arterial pressure. These findings may have important clinical implications. PMID- 2565546 TI - Regional brain glycogen stores and metabolism during complete global ischaemia. AB - Microwave fixation in situ was used to assess regional glycogen and glucose stores in normal rat brain. Glycogen levels were highest in the cerebellum and pons/medulla (38.0 and 35.6 nmol/mg protein), and lowest in the striatum and cerebral cortex (17.4 and 23.6 nmol/mg protein respectively). Glucose concentrations paralleled glycogen, ranging from 5.9 to 10.8 nmol/mg protein. Glycogen, glucose, and lactate were measured during complete global ischaemia (decapitation) to assess regional differences in ischaemic metabolism. Those regions which in normal brain contain higher glycogen and glucose stores were found to maintain significantly higher levels of glycogen and glucose for at least 2 minutes of ischaemia. Lactate accumulated to highest levels after 30 minutes of ischaemia in those regions with highest glucose and glycogen stores. Lactate levels did not, however, rise above 90 nmol/mg protein in any brain region, a level well below that considered potentially neurotoxic. The data indicate considerable regional differences in normal and ischaemic glycogen metabolism that might contribute to known regional differences in vulnerability to global ischaemia. PMID- 2565548 TI - Phosphatases and cathepsin D activities after vasogenic oedema: an experimental study. AB - The role of two phosphatases (acid and alkaline phosphatase) and a lysosomal aspartyl endopeptidase (cathepsin D) in producing rat brain oedema was studied in 3 different rat cerebral areas (i.e. frontal cortex, hippocampus and striatum) at 1, 2 and 3 d after vasogenic brain oedema induction. The percentage of water content in the frontal cortex increased immediately, 1 d after oedema induction and remained high for 2 and 3 d after oedema induction. In the hippocampus and the striatum the water content only increases 3 d after oedema induction. In the oedematous hemisphere (right), when compared to the contralateral hemisphere (left), the acid phosphatase activity decreases in the hippocampus, while the alkaline phosphatase increases in the frontal cortex and striatum; cathepsin D increases only in the striatum. The changes caused by the enzymatic activities were significant only 2 and 3 d after oedema induction. The results of this study show that: (i) the vasogenic oedema induced in experimental conditions was not sufficient to cause a massive liberation of lysosomal enzymes and (ii) brain areas adjacent (below) to the site of the experimental oedematous lesion (frontal cortex) were influenced by oedema induction. PMID- 2565549 TI - Intraventricular infusion of 2-amino-7-phosphonoheptanoate (APH) mitigates ischaemic brain damage. AB - Intracerebroventricular infusion of the competitive N-methyl-D-aspartate (NMDA) antagonist 2-amino-7-phosphonoheptanoate (APH) was evaluated as a neuroprotective regimen in a rat transient cerebral ischaemia model allowing long-term recovery. Ventricular delivery of APH via an osmotic minipump was chosen to allow continuous and direct access to brain tissue of this polar molecule, and because of the potential applicability of such a regimen in clinical situations where the brain is at risk from cerebral ischaemia and a ventricular catheter is in place. The highest tolerable concentration of APH, 50 mM, was given at 1 microliter/h. Selective neuronal necrosis was significantly reduced in the cerebral cortex and the incidence of infarction in the substantia nigra pars reticulata (SNPR) was decreased. A borderline protective effect was seen in the hippocampal CA1 pyramidal neurons, and no significant protection was seen in the caudate nucleus. The results suggest a limited usefulness for APH as a cerebral protective agent. PMID- 2565550 TI - Microvascular and transcranial Doppler sonographic evaluation of cerebral aneurysm flow pattern. AB - Ninety percent of cerebral aneurysms were diagnosed after their first rupture with consecutive subarachnoid haemorrhage. In order to detect the aneurysms before rupture investigation by transcranial Doppler sonography was used on the basis of intra-operative evaluation of the normal aneurysmal flow pattern. In 34 patients the directly registered aneurysmal flow pattern was characterized by mean velocities slower than in parent arteries, by low diastolic flow velocities and by additional peaks superimposed on the normal pulse curve. The transcranial investigation with a large sample volume which includes aneurysm and parent artery was not able to distinguish this aneurysmal flow pattern from the normal one. Only in large to giant aneurysms a pathological flow pattern could be recorded transcranially. PMID- 2565551 TI - Transcranial Doppler in acute brain stem infarction. AB - Two patients with acute brain stem infarctions were examined angiographically and with the transcranial Doppler. One patient had a patent basilar artery but worsened clinically. The second patient had a thrombosis of the left vertebral artery which was dislodged with the angiographic catheter. Thrombolytic therapy was initiated. Serial transcranial Doppler examinations showed persistent patency of the vertebrobasilar system although the patient failed to improve. The usefulness of the transcranial Doppler in the management of acute stroke is discussed in the context of these patients. PMID- 2565552 TI - Effects of thiopentone and etomidate on median nerve somatosensory evoked responses. AB - Somatosensory evoked potentials (SEP) after median nerve stimulation were recorded in 40 patients during infusion of either 15 mg/kg bw thiopentone or 1 mg/kg bw etomidate (n = 10) within 15 min and after 0.3 mg/kg bw etomidate (n = 20). Marked alterations of SEP waveforms and changes in latencies were observed in all patients. Central conduction time (CCT) was significantly correlated to plasma thiopentone concentration. Infusion of high doses of thiopentone and etomidate was followed by a complete loss of middle and long latency components. Amplitude of the primary cortical SEP N20 was found to be unchanged after thiopentone and to be increased after etomidate, indicating the synchronizing properties of this drug. A pronounced increase in SEP latencies and CCT and waveform alterations have to be considered during hypnotic drug administration in intensive care medicine and intraoperatively. PMID- 2565553 TI - Effects of L-carnitine, L-acetylcarnitine and gangliosides on the regeneration of the transected sciatic nerve in rats. AB - Three pharmacological agents, L-carnitine, L-acetylcarnitine and gangliosides, were tested for their ability to enhance the regeneration of the rat sciatic nerve following transection and microsurgical repair. The drugs were administered intraperitoneally at the dose of 50 mg/kg/d for 28 and 56 d postoperatively. At the end of treatment, the motor function recovery of the peroneal component of the sciatic nerve was assessed and the regenerated nerves were analysed morphometrically on histological semi-thin sections. Also, the reinnervated extensor digitorum longus (EDL) muscles were studied histochemically using the adenosine-triphosphatase (ATP-ase) technique 56 d after surgery. Motor function assessment at 56 d after nerve repair revealed that L-acetylcarnitine-treated animals recovered a clinical grade significantly higher (p less than 0.05) than the control animals. Twenty-eight days after nerve repair, the number of myelinated fibres was significantly higher (p less than 0.05) in L acetylcarnitine and ganglioside-treated animals than in control animals. However, 56 d after nerve repair the number of regenerated fibres in all the drug-treated groups was not significantly different from that of the control group. The EDL muscles of the drug-treated animals did not show significant differences from those of control animals with respect to fibre composition and fibre diameter although the L-acetylcarnitine-treated animals exhibited a significantly lower (p less than 0.05) degree of muscle atrophy than did the control animals. The results of the present work seem to indicate that L-acetylcarnitine and to a lesser extent gangliosides exert some favourable effect on the regeneration of the transected sciatic nerve in rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565554 TI - Serum and cerebrospinal fluid enzymes in subarachnoid haemorrhage. AB - Serum levels of creatine kinase (CK) and its isoenzyme CK-MB, lactate dehydrogenase (LDH), hydroxybutyric dehydrogenase (HBDH), glutamic oxalacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) were studied in 50 patients with subarachnoid haemorrhage. In 18 cases the cerebrospinal fluid (CSF) was also examined for total concentration of CK and CK-MB. The results were correlated with the degree of neurological deterioration, the angiographic spasm and prognosis. Concurrent increase of CK-MB, LDH and HBDH serum levels indicates a poor prognosis, whereas increase of GOT and GPT does not have clinical significance. High CK-MB levels in CSF were associated with the worst clinical evolution. However, increase of serum enzymes coincided in most cases with the appearance of the spasm. Monitoring of CK-MB, LDH and HBDH serum levels can be useful for following the evolution of the spasm and in predicting the outcome for patients with subarachnoid haemorrhage. PMID- 2565555 TI - Oestrogen and progesterone sensitivity in cultured meningioma cells. AB - Several studies have detected oestrogen and progesterone receptors in meningioma specimens; recently we have also confirmed the presence of steroid receptors in cultured cells from meningiomas. This paper describes the oestrogen and progesterone receptor assay in cultured cells from 6 meningiomas and the influence of steroid hormones on the cell growth and morphology. Four (66%) of the 6 specimens were positive for both receptors. Growth of cultured cells from tumours without receptors is not appreciably modified by the addition of hormones; the cultured cells from tumours with positive receptors are not essentially influenced by oestrogen, whereas progesterone produces a rapid and marked suppression of the cell growth and modifies their form and adhesivity; also the addition of an oestrogen and progesterone blend produces growth suppression. A similar effect of the progesterone on the cultured cells has also been obtained in a specimen of malignant meningioma. The results of this study suggest that the modulation of progesterone levels may be of therapeutic usefulness, particularly in patients with recurrent malignant meningiomas. PMID- 2565557 TI - A low hippocampal dynorphin A (1-8) immunoreactivity in spontaneously hypertensive rats. AB - Dynorphin A (1-8)-like immunoreactivity (DN-LI A(1-8] was determined by radioimmunoassay in the brains of age matched spontaneously hypertensive rats (SHR) and two normotensive control groups consisting of Wistar-Kyoto (WKY) and Sprague-Dawley (SD) strain rats. A significantly lower DN-LI A(1-8) was found in the hippocampus and hypothalamus of the SHR compared with the WKY groups. DN-LI A(1-8) was 24% of control WKY levels in hippocampus and 79% of that in WKY hypothalamus at 16 weeks. Similar lower levels of DN-LI A(1-8) were also observed in SHR at 4, 8, and 12 weeks during the development of hypertension when compared with both WKY and SD groups. We failed to find significant differences in brain stem DN levels between the groups. The relationship between the low hippocampal dynorphin levels in SHR and the hypertension is problematical because the differences were present before (4 wks), during (8 and 12 wks) and after (16 wks) its development. PMID- 2565556 TI - Long-term haloperidol treatment decreases somatostatin binding in rat brain. AB - The effects of short and long-term haloperidol treatment on somatostatin concentration and specific binding in rat cerebral cortex and hippocampus were examined using the binding ligand 125I-Tyr1-somatostatin. Haloperidol treatment did not affect the concentration of somatostatin-like immunoreactivity in the two brain areas. Nevertheless, long-term, and not short-term, haloperidol treatment decreased the number of somatostatin receptors in the cerebral cortex and hippocampus. No significant differences in the apparent binding affinity values were seen after haloperidol treatment. When added at the time of the binding assay haloperidol 34.2 microM produced a 42% and 27% decrease in cerebrocortical and hippocampal membrane somatostatin receptors respectively. PMID- 2565558 TI - Measurement and chromatographic characterization of prodynorphin-derived peptides in the guinea-pig ileum. AB - Guinea-pig ileum was dissected and the mucosa, submucosa and external musculature extracted with aqueous acetic acid for measurement of four prodynorphin-derived peptides, namely dynorphin A 1-8, dynorphin A 1-17, dynorphin B, and alpha neoendorphin. The peptide-like immunoreactive material extracted from the external musculature was characterized by multi-dimensional chromatographic analysis and compared to synthetic porcine standards. The chromatographic methods utilized were: reversed-phase high performance liquid chromatography (RP-HPLC), using two different eluants; cation exchange high performance liquid chromatography (CE-HPLC) and gel filtration chromatography. The dynorphin A 1-8 like immunoreactive material was homogeneous and coeluted with the standard in all chromatographic modes. The dynorphin A 1-17-like and dynorphin B-like immunoreactive material was heterogeneous but showed a peak that coeluted with synthetic standard in all chromatographic modes. The alpha-neoendorphin-like immunoreactive material also appeared to be heterogeneous with the major component on CE-HPLC coeluting with the synthetic peptide standard while the major component on RP-HPLC eluted differently. It was concluded that the guinea pig ileum contains immunoreactivity for peptides derived from all coding regions of the prodynorphin gene and that these peptides may be present in multiple immunoreactive forms. PMID- 2565559 TI - Memory systems in the chick: dissociations and neuronal analysis. AB - Young domestic chicks learn to recognize the visual characteristics of an object to which they are exposed. A restricted part of the forebrain, the intermediate and medial part of the hyperstriatum ventrale (IMHV) is implicated in this process. This form of exposure learning can be dissociated from (i) the ability to learn certain procedures or skills, and (ii) a predisposition to attend to particular types of naturalistic objects. The first of these dissociations is reminiscent of that found in certain human organic amnesias, whilst the second may have its counterpart in the processes involved in face recognition by infants. The right and left IMHV play different roles in the memory that underlies imprinting. The cellular and molecular processes involved in this form of memory are discussed. PMID- 2565560 TI - Ontogenesis of tyrosine hydroxylase-immunopositive structures in the rat hypothalamus. An atlas of neuronal cell bodies. AB - The development of the catecholaminergic system in the hypothalamus and in the septal region was studied in rats from the 12th fetal day until the 9th postnatal day. Catecholaminergic structures were visualized with pre-embedding immunocytochemistry using antiserum to tyrosine hydroxylase. An intensification of diaminobenzidine product with silver and gold was additionally applied to make the immunocytochemical technique more sensitive. In this paper only the data on the appearance and distribution of the tyrosine hydroxylase-immunopositive neurons (cell bodies) are presented, whereas the catecholaminergic innervation of the hypothalamus with the tyrosine hydroxylase-immunopositive fibers is the topic of an accompanying paper. Sparse tyrosine hydroxylase-immunopositive neurons were first observed in the anlage of the hypothalamus and septal region on the 13th fetal day. Their number increased progressively with age and by the 15th fetal day they already gave rise to a large dorsal accumulation. From the 18th fetal day on, tyrosine hydroxylase immunopositive neurons began to occupy their definitive positions, mainly concentrating within the hypothalamus: in the zona incerta, periventricular and arcuate nuclei. To a lesser extent, they were concentrated in the medial preoptic area, suprachiasmatic, supraoptic, paraventricular, dorsomedial, and anterior hypothalamic nuclei. The data on the distribution of the tyrosine hydroxylase-immunopositive neurons both in the hypothalamus and in the septal region during ontogenesis are summarized in the precise atlas. Primarily small bi- and unipolar catecholaminergic neurons first observed in the youngest fetuses undergo cytodifferentiation during ontogenesis, giving rise to at least two different populations localized ventrally, mainly in the arcuate nucleus, and dorsally, in the zona incerta. The neurons of the former population remain similar to those of the youngest fetuses, whereas the neurons of the latter increase significantly in size, forming several long, highly ramified processes. PMID- 2565561 TI - Ontogenesis of tyrosine hydroxylase-immunopositive structures in the rat hypothalamus. Fiber pathways and terminal fields. AB - The innervation of the hypothalamus and septal region by catecholaminergic fibers was studied in rats from the 12th fetal day until the 9th postnatal day. Catecholaminergic fibers were visualized with preembedding immunocytochemistry using antibodies to tyrosine hydroxylase. An intensification of diaminobenzidine product with silver and gold was additionally applied to increase the sensitivity and resolution power of the routine immunocytochemical technique. It has been demonstrated that, from the 13th fetal day, the hypothalamus and the septal region receive catecholaminergic fibers either belonging to the hypothalamic neurons or coming with the medial forebrain bundle from the outside of the hypothalamus. As the development of the hypothalamus proceeds, these fibers form the extensive networks within some neurosecretory centers either containing (the zona incerta, periventricular nucleus, etc.) or almost lacking (suprachiasmatic and paraventricular nuclei) the catecholaminergic neurons. In the former case, they terminate on the processes or perikarya of catecholaminergic neurons, while in the latter case their varicosities surround the immunonegative presumptive neurons in a basket-like manner. Moreover, from the 18th fetal day catecholaminergic fibers penetrate between the ependymal cells towards the 3rd ventricle and the primary capillary plexus of the hypophysial portal circulation, apparently providing the release of catecholamines to the cerebrospinal fluid and portal blood, respectively. The data obtained in this study are considered as the morphological basis for the involvement of the hypothalamic catecholamines in neuroendocrine regulations during ontogenesis. PMID- 2565562 TI - Stimulation of the nucleus basalis of Meynert increases cerebral cortical blood flow in rats. AB - Focal electrical stimulation of the magnocellular nucleus of the basal forebrain (nucleus basalis of Meynert; NBM) or a microinjection of L-glutamate (50 nmol) into the NBM increased cerebral cortical blood flow in the parietal lobe in urethane-anesthetized rats. The vasodilative responses were elicited only ipsilateral to the site of stimulation. Most of the vasodilative responses were abolished by intravenous administrations of muscarinic and nicotinic cholinergic blocking agents (atropine 0.5 mg/kg and mecamylamine 2 mg/kg). This suggests that the cholinergic projecting system sending fibers from the NBM to the parietal lobe contributes to the vasodilation of the cortex by activating muscarinic and nicotinic cholinergic receptors. PMID- 2565563 TI - Stimulation of the nucleus basalis of Meynert increases acetylcholine release in the cerebral cortex in rats. AB - The effect of focal stimulation of the magnocellular nucleus of the basal forebrain (nucleus basalis of Meynert; NBM) on acetylcholine (ACh) release in the cerebral cortex in the parietal lobe was examined in halothane-anesthetized rats. ACh was measured using a microdialysis method. Focal electrical stimulation of the unilateral NBM increased ACh release in the ipsilateral cerebral cortex in stimulus intensity and frequency dependently. Microinjection of L-glutamate (100 nmol) into the unilateral NBM also increased ACh release in the ipsilateral cerebral cortex. The ACh release in the contralateral cerebral cortex was not affected by these unilateral stimulations of the NBM. It was concluded that focal stimulation of the NBM releases ACh from cortical terminals of cholinergic fibers originating in the NBM. PMID- 2565564 TI - Glutamate receptor agonists increase intracellular Ca2+ independently of voltage gated Ca2+ channels in rat cerebellar granule cells. AB - Changes in membrane potential and cytosolic free Ca2+ concentrations, [Ca2+]i, in response to L-glutamate and glutamate receptor agonists were measured in rat cerebellar granule cells grown on coverslips. The membrane was depolarized by the application of L-glutamate and kainate, and by elevating the extracellular K+ concentration, as determined by using the membrane potential probe bisoxonol (DiBA-C4-(3)). The [Ca2+]i as measured with fura-2 was 220 nM on average under resting conditions and increased by raising the extracellular K+ and by applying L-glutamate, kainate, quisqualate or N-methyl-D-aspartate (NMDA). Verapamil and nifedipine reduced the high-K+ induced rise in [Ca2+]i but did not significantly affect the responses produced by NMDA, quisqualate and kainate, suggesting that the increase in intracellular Ca2+ in response to glutamate receptor agonists is primarily due to Ca2+ influx through receptor-coupled ion channels. PMID- 2565565 TI - Reversal of desipramine-induced suppression of paradoxical sleep by a long-acting somatostatin analogue (octreotide) in rats. AB - EEG sleep recordings were performed in rats under intraperitoneal injections of saline, desipramine (DMI, 4 mg/kg) an inhibitor of noradrenaline reuptake, and DMI plus the octapeptide somatostatin analogue (octreotide, 0.2 mg/kg). As already reported, DMI resulted in selective suppression of paradoxical sleep (PS) and increased slow wave sleep (SWS). The administration of the octapeptide somatostatin analogue totally reversed the DMI-induced suppression of PS, but had no effect on SWS. This finding confirms previous results demonstrating a role of somatostatin in the generation of PS. In addition, it suggests that the suppression of PS by DMI may be due to an inhibitory effect on somatostatin release, rather than to an alteration of brain noradrenaline. PMID- 2565566 TI - REM sleep without atonia after lesions of the medial medulla. AB - Rapid eye movement (REM) sleep is normally accompanied by a complete suppression of tone in the antigravity musculature. Pontine lesions have been shown to block this suppression, producing a syndrome of REM sleep without atonia. We now report that glutamate-induced lesions of the medial medulla, including the nucleus magnocellularis, caudal nucleus gigantocellularis and rostral nucleus paramedianus, produce REM sleep without atonia. These nuclei may function as part of a ponto-medullary system suppressing muscle tone in REM sleep. PMID- 2565567 TI - Modification of potassium-induced interictal bursts and electrographic seizures by divalent cations. AB - Reduction of external calcium and magnesium from 1.5 to 1.2 mM intensified potassium-induced interictal bursts, increased the likelihood of electrographic seizure occurrence in CA1, and rendered seizure initiation independent of N methyl-D-aspartate (NMDA) receptor activation. In contrast to slices bathed in 1.5 mM divalent cations, in 1.2 mM divalents spontaneous CA1 seizures still occurred in CA1 minislices that contained at least 1500 neurons after removal of the CA3 burst generator, suggesting that divalent cations critically modulate the dependence of CA1 seizure initiation on interictal input. Since this slight reduction in external divalent cations enhanced tissue excitability, similar changes might promote epileptogenesis in situ. PMID- 2565568 TI - K+ differentially affects the excitatory amino acids- and carbachol-elicited inositol phosphate formation in rat brain synaptoneurosomes. AB - K+, excitatory amino acids (EAAs) and carbachol (Carb) were tested separately or in pairs for their ability to stimulate inositol phosphate (IPs) formation in rat forebrain synaptoneurosomes. K+ ions per se, stimulate IPs synthesis (158% of the control value) as well as EAAs and Carb. The glutamate (Glu)- and quisqualate (QA)-elicited IPs formation is not additive with that evoked by K+. Inversely, K+ ions (up to 30 mM) potentiate the Carb-induced IPs accumulation. These results indicate that QA (or Glu) and Carb enhance IPs formation independently and that QA- and K+ -induced IPs responses are interdependent. This suggests that they share a 'common intermediate' step in the multistep mechanism which leads from receptor activation to the IPs synthesis. This 'common intermediate' step may be depolarization and/or Na+ influx. PMID- 2565569 TI - The psychoneuroendocrinology of functional hypothalamic amenorrhea. AB - Women with functional hypothalamic amenorrhea display multiple neuroendocrine aberrations suggestive of altered central neurotransmission. The role of antecedent stress as an explanation for both the neurochemical changes and the dysfunctional behavior of these women is examined by utilizing concepts provided by the animal model of inescapable shock and the human condition of post traumatic stress disorder. PMID- 2565570 TI - Reduction of the effects of growth hormone release inhibiting factor enhances plasma growth hormone response to GHRH. AB - Responses of serum growth hormone (hGH) to glucagon (G), growth hormone releasing hormone (GHRH) and G/GHRH were measured in 8 normal adults and 6 patients with growth hormone deficiency (GHD). In normal adults, serum hGH reached its peak value (12.7 +/- 1.6 ng/ml) at 150 +/- 10 min, as blood glucose declined to its minimum after a transitory hyperglycemia in G test. The normal adults were responsive to GHRH test (GH peak 14.7 +/- 2.3 ng/ml at 30 +/- 0 min). In GHD, the responders to both G and GHRH tests showed a strongly positive response in G/GHRH test, with a serum hGH peak value of 34.6 +/- 4.1 ng/ml at 131 +/- 8 min being much higher than that of either single G or GHRH test (P less than 0.01), but without significant difference to the sum of the two single tests (P greater than 0.10). Among GHD patients, only 2 responded to GHRH and G/GHRH tests with hGH peak values 6.8 +/- 0.7 and 6.9 +/- 0.7 ng/ml at 45 +/- 15 and 90 +/- 0 min, respectively, both peak values being essentially similar (P greater than 0.10). We suggest that the mechanism of stimulation of pituitary hGH secretion in G test might involve inhibition of release of hypothalamic GH release inhibiting factor (GHRIF) caused by hypoglycemia after a transitory hyperglycemia following G injection. These results may further confirm our previous postulation (1986) that insulin hypoglycemia may increase hGH release by inhibiting hypothalamic cell secretion of GH release inhibiting factor. PMID- 2565571 TI - The award of medals by the president, Sir George Porter, at the anniversary meeting, 30 November 1988. PMID- 2565572 TI - Differential response to a competitor by Atlantic salmon adopting alternative life-history strategies. AB - The life cycle of the Atlantic salmon is extremely variable. In good growing conditions, juvenile salmon either metamorphose into the migratory phase by their second spring, or delay this for at least another year. The strategy appears to be decided in their first summer. This study compared competitive responses of fish adopting the two strategies. Laboratory experiments showed that the two types of fish had similar foraging efficiencies in isolation. However, although a simulated competitor had little effect on the feeding behaviour of fast developing fish, it caused an 18-fold increase in the incidence of failed feeding attempts by fish delaying development. The probability of an attack failing was dependent on how close the competitor came. PMID- 2565573 TI - Non-mitochondrial calcium ion regulation in rat ventricular myocytes. AB - Ca2+ exchange has been measured in a suspension of rat ventricular myocytes treated with digitonin or saponin to render the sarcolemma permeable to small molecules and ions. Two fractions of exchange were identified, one that was attributed to the mitochondrial component of the cell and the other to a non mitochondrial fraction. Mitochondrial Ca2+ uptake was blocked by sodium azide and depended on respiratory substrates whereas non-mitochondrial uptake occurred independently of these molecules but was dependent on ATP and creatine phosphate. Non-mitochondrial Ca2+ uptake could be induced at a Ca2+ concentration below 1 microM and the initial rate increased with concentration up to 100 microM. Uptake could be reversed by sulmazole (a caffeine-like substance) and this reversal in turn inhibited by ryanodine. These properties suggest that the major locus for non-mitochondrial Ca2+ exchange is at the sarcoplasmic reticulum. Ca2+ exchange could be modulated by a number of agents, including carnosine, but was unaffected by others, including Na+, inositol trisphosphate and cyclic AMP. A kinetic model of the data is presented, which incorporates similar data of Ca2+ uptake into the mitochondrial fraction. The rates of Ca2+ exchange measured in these experiments suggest that these two components of the cell can reduce the sarcoplasmic Ca2+ concentration rapidly enough to account for the observed transient nature of the isometric twitch. Furthermore, it is suggested that both non-mitochondrial and mitochondrial fractions of the cell could significantly contribute to tension relaxation in rat cardiac muscle. PMID- 2565574 TI - Early social status and the development of life-history strategies in Atlantic salmon. AB - Atlantic salmon have a variable life cycle. In good growing conditions, underyearling fish may metamorphose into the migratory smolt phase during their second spring, or delay at least a further year. The strategy adopted by particular fish appears to become fixed during their first summer. This paper examines whether either feeding efficiency or dominance in mid-summer correlates with the life-history strategy adopted. Eighty fish were individually marked and their feeding efficiency (= mean handling time for food items) and dominance rank measured under laboratory conditions in mid-July. Growth rates of the fish were then monitored over the next three months, until developmental strategies became apparent. Discriminant and logistic regression analyses revealed that both dominance rank and size attained by July were independent, significant predictors of future developmental pattern (the age at metamorphosis being correctly predicted on the basis of rank and size in 84% of cases) whereas feeding efficiency had no effect. Thus fish that were dominant or larger two months after first feeding or both had a greater probability of migrating after only one year in freshwater than those more subordinate or smaller or both. PMID- 2565575 TI - Automated site-directed drug design: a general algorithm for knowledge acquisition about hydrogen-bonding regions at protein surfaces. AB - This is the first of four papers that begin to explore the possibility of automated site-directed drug design. A general outline is given of the logical steps involved in approaching the problem. The starting point is the process of knowledge acquisition about the site. An algorithm is described here for the construction of a map of hydrogen-bonding regions at protein surfaces directly from the Brookhaven Protein Data Bank coordinates. Hydrogen-bonding atoms are located, intramolecular bonds are searched for, hydrogen-bonding atoms at the surface are found and hydrogen-bonding regions are computed at the accessible surface. A grid is placed within each region discovered and the probability of hydrogen bonding at each grid point is computed. The output of the program is a map of hydrogen-bonding regions displayed within a user-defined window. This information can be used as part of a knowledge base for the automatic construction of novel ligands to fit specified binding sites. PMID- 2565576 TI - Automated site-directed drug design: the prediction and observation of ligand point positions at hydrogen-bonding regions on protein surfaces. AB - The HSITE program proposed in the previous paper was written to define putative ligand-point regions that could be found at protein surfaces. These regions would represent positions for hydrogen-bonding acceptor and donor atoms. In this paper the prediction of the location of these regions is compared with: (1) the position of the oxygen atoms of water molecules on the hydrated proteins myoglobin and plastocyanin; and (2) the position of hydrogen-bonded atoms in methotrexate and NADPH co-crystallized with dihydrofolate reductase, and in amidinophenyl-pyruvate co-crystallized with trypsin. The prediction of ligand point regions is in agreement with the surveys of experimental data for water molecule positions in protein crystals and with the positions of hydrogen-bonding atoms found in co-crystallized ligands. PMID- 2565577 TI - Automated site-directed drug design: the concept of spacer skeletons for primary structure generation. AB - This paper examines the problem of automated structure generation at specified binding sites. The objective is to obtain molecular graphs that span the binding site and incorporate predicted ligand points at their vertices. Three approaches are considered: brute-force techniques, subgraph addition and spacer skeletons. Spacer skeletons are assemblies of molecular subgraphs and are used to reduce the combinatorial problems of structure generation to a practicable level for future analysis. This description is restricted to structure generation in two dimensions. Assemblies of rings are examined for planarity by searching the Cambridge Structural Database. Appropriate spacer skeletons may then be fitted to arrays of site points. PMID- 2565578 TI - Automated site-directed drug design: the formation of molecular templates in primary structure generation. AB - In this paper the spacer skeleton concept is used to produce molecular graphs of putative ligands for binding sites. The skeletons are transformed into molecular templates within the constraints of the accessible surface of the ligand-binding site. A distance-matrix method is used to compare ligand points with vertices of the spacer skeleton through a permutation of all possible correspondences. A tolerance parameter is used to screen for poor matches. As a result, a small number of matched vertices and ligand points are produced. These are fitted into the site by a constrained optimization routine using an analytical function. Ligand points fall within the site and are optimally positioned adjacent to the corresponding site points; other vertices of the spacer skeleton lying beneath the accessible surface of the site are clipped off. A molecular template is thereby formed with its vertices linked to the ligand points. The final step is to verify that the bonding integrity of the skeleton remains. The computational methods outlined in this paper have been tested at two binding sites: the pteridine binding site in dihydrofolate reductase and the amidinophenylpyruvate site of trypsin. Molecular graphs for both sites were generated automatically; they showed strong similarity to those of the natural ligands. PMID- 2565579 TI - Properties of membrane ion conductances evoked by hormonal stimulation of guinea pig and rabbit isolated hepatocytes. AB - Membrane conductance changes evoked in isolated guinea-pig or rabbit hepatocytes by hormonal stimulation were studied with the whole-cell patch clamp technique. In Cl-containing solutions, noradrenaline (NA), ATP or angiotensin II (AII) evoked an increase of conductance to both K (GK) and Cl (GCl) ions. Activation of GK occurred after a delay of several seconds and was sustained in the presence of hormone. Activation of GCl was transient, lasting several seconds, and arose either at the same time or shortly after the increase in GK. Conductances showed an initial rapid rise and slow oscillatory changes during maintained hormone application. The NA-induced current reversed at -19 mV in Cl solutions, between the equilibrium potentials for chloride (ECl = 0 mV) and potassium ions (EK = -85 mV), and at -75 mV, near EK, in Cl-free solution. In both conditions whole-cell current-voltage curves were linear in the range -100 mV to +40 mV. The conductance increase produced by NA to Cl- ions was about 50 nS, that to K+ ions was 6 nS. The potassium conductance increase was abolished by the polypeptide toxin apamin (50 nM). An increase in membrane current noise was associated with NA-evoked outward K+ current and blocked by apamin. Spectral analysis gave estimates of the elementary K channel conductance of 1.7 pS. Power spectra were fitted by two Lorentzian components, with average half-power frequencies of 2 and 190 Hz. These results are discussed in relation to the single-channel properties and indicate that the open probability of K channels during the NA response is high. In Cl solutions, with apamin to block the K conductance, no increase in current noise was detected during the large Cl conductance evoked by NA. This suggests either that Cl channels are of very low unitary conductance (less than 1 pS) or that Cl transport is due to a membrane carrier. The complex time-course of hormonally evoked conductances is not due to the properties of ion conductances per se but probably to underlying changes of intracellular second-messenger concentration. PMID- 2565580 TI - Novel neurotransmitters in the autonomic nervous systems of nonmammalian vertebrates. PMID- 2565581 TI - Somatostatin and its analogues in the therapy of gastrointestinal disease. PMID- 2565582 TI - Placebo-controlled study on acute and subchronic effects of buspirone vs bromazepam utilizing psychomotor and cognitive assessments in healthy volunteers. AB - The acute and subchronic effects of orally administered buspirone (acute: 15 mg; subchronic: 5 mg t.i.d.; Day 7:5 mg) and bromazepam (acute: 6 mg; subchronic: 2 mg t.i.d.; Day 7: 2 mg) on psychomotor and cognitive parameters were explored vs placebo in a randomized, double-blind crossover design with three periods in 12 healthy male volunteers. The washout periods between active treatments lasted one week. Psychomotor and cognitive measurements were taken with the Oculodynamic Text (ODT), which is a computerized psychophysiological online testing device for the simultaneous measurement of electrooculographic and performance (signal identification) parameters. Subjective side effects were recorded with a symptom checklist (41 items) with regard to the first and seventh day of administration. Intradiurnal assessments were done at 0, +1, +3, and +5 hours post-dose on Days 1 and 7 of each of the three randomized treatment periods. Buspirone behaved placebo-like in the EOG and signal-identification parameters after acute and subchronic administration. After bromazepam the respective parameters were significantly impaired--acutely as well as subchronically. Acute and subchronic buspirone was significantly superior to bromazepam with regard to the majority of psychophysiological parameters. The incidence of acute side effects was higher for both active treatments than for placebo. With buspirone there were fewer acute and subchronic side effects than with bromazepam. The side effect profiles of the three preparations were different. With regard to all active treatments the incidence of side effects was lower on Day 7 after the subchronic dosing period--with a third of the subchronic dose--than after the first single administration. PMID- 2565583 TI - Efficacy and tolerability of a new antipsychotic compound (savoxepine): results of a pilot-study. AB - Savoxepine is a new tetracyclic compound displaying potent neurolepticlike effects in pharmacological studies. Of particular interest is its preferential binding to dopamine-2 receptors in the hippocampus, which leads to the hypothesis that savoxepine may exert antipsychotic effects at doses not inducing extrapyramidal side-effects. In an open pilot-study 18 patients suffering from acute schizophrenic psychoses or paranoid syndromes were treated with savoxepine in an individually adapted dose range from 0.50 to 10 mg per day. A good antipsychotic efficacy could be demonstrated in 10 of 16 patients. Savoxepine was found to be generally well tolerated. Contrary to expectations, mild or moderate extrapyramidal side-effects, especially of the parkinsonian type, were registered. Future research has to test the suggested advantage of savoxepine in comparison with other neuroleptic drugs. PMID- 2565584 TI - Cyclic AMP oscillations in suspensions of Dictyostelium discoideum. AB - A model developed previously for signal relay and adaptation in the cellular slime mould Dictyostelium discoideum is shown to account for the observed oscillations of calcium and cyclic AMP in cellular suspensions. A qualitative argument is given which explains how the oscillations arise, and numerical computations show how characteristics such as the period and amplitude of the periodic solutions depend on parameters in the model. Several extensions of the basic model are investigated, including the effect of cell aggregation and the effect of time delays in the activation and adaptation processes. The dynamics of mixed cell populations in which only a small fraction of the cells are capable of autonomous oscillation are also studied. PMID- 2565585 TI - A double-blind trial of essential fatty acid supplementation in patients with tardive dyskinesia. AB - This study reports the results of a trial of essential fatty acid (EFA) supplementation in psychiatric patients (predominantly schizophrenics) with movement disorders. Evidence of EFA deficiency in these patients was observed. The antidyskinetic effect of EFA supplementation was marginally significant but not clinically important. However, active treatment produced highly significant improvements in total psychopathology scores and schizophrenia subscale scores, and a significant improvement in memory. PMID- 2565586 TI - Neuroleptic-induced akathisia: a review. AB - Neuroleptic-induced akathisia (NIA) is a relatively common side effect of neuroleptics, in which patients complain of a subjective sense of restlessness usually referable to the legs and have characteristic motor movements. This paper will review: 1) history of spontaneously occurring syndromes of pathologic restlessness and NIA, 2) the clinical significance of NIA, 3) issues concerning the diagnosis and quantification of NIA, 4) treatments of NIA and 5) possible future directions for research in this area. Special attention will be paid to newer treatments for this syndrome, specifically beta-blockers. PMID- 2565588 TI - An alpha 2 adrenoceptor antagonist, Org 3770, enhances nocturnal melatonin secretion in man. AB - Nocturnal plasma melatonin concentration was significantly increased in ten normal volunteers following the administration of an alpha 2 adrenoceptor antagonist, Org 3770 (30 mg). This result supports the existence of an alpha 2 adrenergic influence on melatonin secretion in man and provides a possible clinical neuroendocrine marker of the action of an alpha 2 antagonist. PMID- 2565587 TI - Effects of centrally acting beta adrenergic agonists on discrete trial conditioned avoidance behavior in rats. AB - The acute effects of centrally acting beta adrenergic agonists on discrete trial conditioned avoidance responding in rats were examined. Clenbuterol (0.01-3.0 mg/kg), salbutamol (0.01-30 mg/kg), and prenalterol (30-300 mg/kg) suppressed avoidance responding in a dose-dependent manner at doses that did not produce escape failures. For comparative purposes, the effects of the tricyclic antidepressant desipramine (1.0-30 mg/kg) and the antipsychotic haloperidol (0.03 0.3 mg/kg) were similarly assessed. Both compounds suppressed avoidance responding in a dose-dependent manner. Only haloperidol (0.3 mg/kg) produced escape failures. Administered alone, the beta adrenergic antagonist propranolol (1.0 and 10 mg/kg) did not affect avoidance behavior. When administered prior to clenbuterol (0.1 mg/kg), salbutamol (1.0 mg/kg), or prenalterol (100 mg/kg), propranolol antagonized the beta adrenergic agonist-induced suppression of avoidance responding. The suppressive effect of desipramine (3.0 mg/kg) on avoidance performance only tended to be attenuated by propranolol. Propranolol had no effect on the ability of haloperidol (0.1 mg/kg) to reduce avoidance responding. These results suggest that the effects of the beta adrenergic agonists clenbuterol, salbutamol, and prenalterol on discrete trial avoidance behavior are mediated, in part, through agonist interactions with beta adrenergic receptors. PMID- 2565589 TI - Psychological and immunological predictors of genital herpes recurrence. AB - The relationships among stressful life experience, mood, helper-inducer (CD4+) and suppressor-cytotoxic (CD8+) T cells and genital herpes simplex virus (HSV) recurrence were investigated prospectively in 36 patients with recurrent HSV. The following factors were measured monthly for six months: stressful life experience (including current acute and ongoing stressors, residual effects of previous stressors, and anticipation of future stressors), negative mood, health behaviors, other possible HSV triggers, HSV recurrences, and the proportion of CD4+ and CD8+ cells (in half the sample). Results averaging monthly scores over the six-month study period indicated that: 1) subjects with high levels of stressful experience had a lower proportion of both CD4+ and CD8+ cells, 2) subjects with high levels of depressive mood, anxiety, or hostility had a lower proportion of CD8+ cells, and 3) subjects with high levels of depressive mood who did not report many symptoms of other infections had a higher rate of HSV recurrence. A model is proposed linking depressive mood, CD8+ cells, and HSV recurrence. PMID- 2565590 TI - [A study of prescriptions for psychotropic drugs at a French psychiatric hospital]. AB - The prescribing habits were studied in a psychiatric hospital which provides services for one district of Paris. The sample consisted of 175 inpatients. A mean of 2.1 psychotropic drugs was prescribed per patient, and more than two thirds received two psychotropic drugs or more. Almost one-half had a PRN order. Major tranquilizers were frequently used in patients not having a psychotic disorder and were associated to anticholinergic drugs in 55% of the cases. Factors associated with drug prescriptions were diagnosis, age and unit of treatment. Our results are similar to those of previous studies on excessive polypharmacy. The discussion draws attention on the need to obtain data on the rationale for prescribing and the response to treatment. Emphasis is placed on the potential usefulness of studying drug prescriptions for psychiatric practice. PMID- 2565591 TI - [Mixed benzodiazepine poisoning and reversal with flumazenil (Ro 15-1788)]. AB - A case of 17 year old female with acute benzodiazepines (800 mg of bentazepan and 400 mg de chlordiazepoxide) and tricyclic antidepressants (500 mg of imipramine) self poisoning is reported. Initial examination showed comatous patient (Glasgow coma scale 7, sedation scale 4) and a critical respiratory failure with a suspicious of pulmonary aspiration of gastric contents. Supportive measures were started and flumazenil administered intravenous (0.1 mg/min) until a total dose of 1.2 mg; 15 minutes later neurological state was absolutely normal. We conclude that flumazenil is a selective and useful benzodiazepine antagonist in the treatment of acute benzodiazepine poisoning. PMID- 2565592 TI - [Therapeutic application of anesthetic blocks in weever-fish stings]. AB - Five cases of weaver-fish sting (weaver Trachinus) are described. All patients presented severe pain and peripheral vascular disorders. In 4 cases the sting was in the upper limb and treated with brachial plexus axillary block. One patient had been stung in the foot and treated with epidural continuous block with catheter: In all cases bupivacaine 0.5% was employed and evolution was good 24-48 hours later. We conclude that anesthetic blocks should be included in the symptomatic treatment of those cases, that is: steroids, antihistaminics, calcium, analgesic agents, antibiotics, and antitetanic vaccine. PMID- 2565593 TI - [Metastasis of carcinoma of the colon to cryptorchid testis. An infrequent finding]. PMID- 2565594 TI - Effects of d- and l-pentazocine on the release and uptake of norepinephrine in rat brain cortex. AB - Effects of d- and l-pentazocine on the release and uptake of norepinephrine and other neurotransmitters were examined in rat cerebral cortex. D- and l pentazocine (10(-6)-10(-4) M) evoked the release of tritium from rat cortex slices preloaded with [3H]norepinephrine. Both isomers of pentazocine (3 x 10(-5) M) also evoked the release of tritium from slices preloaded with [3H]dopamine and [3H]5-hydroxytryptamine ([3H]5-HT) but did not from those preloaded with [3H] gamma-aminobutyric acid ([3H]GABA) and [3H]choline. The releasing effect of pentazocine was neither dependent on the extracellular calcium nor antagonized by naloxone. Both isomers inhibited the uptake of [3H]norepinephrine, [3H]dopamine and [3H]5-HT but not that of [3H]GABA and [3H]choline into synaptosomes prepared from the cortex. These results suggest that the effects of d- and l-pentazocine on release and uptake are relatively specific to monoamines and might explain some aspects of pharmacological actions of pentazocine. PMID- 2565595 TI - [Systemic non-rheumatoid inflammatory arthropathies or connective tissue diseases]. PMID- 2565596 TI - Daytime naps in narcoleptic patients. PMID- 2565597 TI - Utilization of amino acids and peptides by Fusobacterium nucleatum. AB - Fusobacterium nucleatum strains ATCC 10953, Fevl, F1, F3, and F6 utilized amino acids, in particular glutamate, histidine, and aspartate were common to all strains. Strain differences were observed in the utilization of threonine, serine, lysine, tyrosine, and methionine, and only strain ATCC 10953 utilized all these amino acids. The glutamate and histidine pools were in all cases fully depleted before the other amino acids were attacked and at the same time all strains except 10953 started to utilize peptides at a noticeable rate. For strain Fevl, glutamyl- and aspartyl-containing peptides seemed to be of considerable nutritional importance, and this strain did not grow on a medium based on amino acids alone. On the other hand, strain 10953 did not utilize any peptides to a noticeable extent, and it could grow on an amino acid based medium. PMID- 2565598 TI - [Coronary steal syndrome following internal mammary artery bypass surgery: incidence and recommendations for avoiding it]. AB - With the increased use of the internal mammary artery for coronary artery bypass surgery, new complications are seen. This report describes the twelfth case of a coronary-subclavian steal syndrome. It summarizes all previously reported cases and makes some recommendations in order to avoid and treat this condition. PMID- 2565600 TI - [Roentgen diagnosis of the foot]. AB - Diagnostic modalities in trauma studies of the foot include conventional radiography, ultrasound, computed tomography (CT), magnetic resonance imaging (MRI), and digital projection radiography (DR). Several projections and special views of the conventional radiographic technique are shown with special respect to the hindfoot and the subtalar joint. The importance of ultrasound for soft tissue diagnosis is emphasized. Other modalities (CT, MRI, DR) are illustrated. PMID- 2565599 TI - Access to a messenger RNA sequence or its protein product is not limited by tissue or species specificity. AB - RNA amplification with transcript sequencing (RAWTS) is a rapid and sensitive method of direct sequencing that involves complementary DNA synthesis, polymerase chain reaction (PCR) with a primer or primers containing a phage promoter, transcription from the phage promoter, and reverse transcriptase-mediated sequencing. By means of RAWTS, it was possible to sequence each of four tissue specific human messenger RNAs (blue pigment, factor IX, phenylalanine hydroxylase, and tyrosine hydroxylase) in four cell types examined (white blood cells, liver, K562 erythroleukemia cells, and chorionic villus cells). These results indicate that there is a basal rate of transcription, splicing, and polyadenylation of tissue-specific mRNAs in adult and embryonic tissues. In addition to revealing sequence information, it is possible to generate a desired in vitro translation product by incorporating a translation initiation signal into the appropriate PCR primer. RAWTS can be used to obtain novel mRNA sequence information from other species as illustrated with a segment of the catalytic domain of factor IX. In general, the ability to obtain mRNA sequences rapidly across species boundaries should aid both the study of protein evolution and the identification of sequences crucial for protein structure and function. PMID- 2565601 TI - [Reconstructive foot surgery following complex trauma of the foot]. AB - Compound post-traumatic foot deformity may follow indirect, direct, or combined trauma. This is demonstrated by a number of case reports. Particularly during adolescence, primary injuries to the soft tissue, with neurovascular injury or compartment syndrome, and post-traumatic skin contractures can cause severe growth deformity at the site of the injury and distal to it. Compression screw arthrodesis, using the 6.5-mm cancellous screw with the triple arthrodesis and the 3.5-mm cortical screw with the Lisfranc arthrodesis, has particular significance. It is stable, it allows functional treatment later, and bone union takes place rapidly. The most important surgical principles in post-traumatic foot surgery are: precise reconstruction of the foot axes, the medial and lateral foot length, and the longitudinal and transverse arches, i.e., restitution of the normal foot anatomy. PMID- 2565602 TI - [Technics of primary and secondary amputation of the foot]. AB - Maintenance therapy of the traumatized foot by partial amputation, in contrast to lower-leg amputation with subsequent artificial leg adjustment, should always be attempted if there is any possibility of shaping a stump that is independent of the prosthesis. As in former years, the disarticulation technique is performed; however, today the technique has been modified to obtain more functional results and to conserve tissue, which may mean that transosseous amputation is practicable. The quality of the soft-tissue flap and stability are the determining factors in the quality of the stump. These prerequisites mean that the operative technique must be adapted to the requirements of the patients; the soft tissue must be preserved and procedures used that will permit reconstructive procedures later. PMID- 2565603 TI - Tumor suppressor genes and inherited predisposition to malignancy. PMID- 2565604 TI - The neu (c-erbB-2) oncogene. AB - The neu gene was first identified in rat tumors that had been induced by the carcinogen ethyl nitrosourea. The human homolog of neu, usually designated c-erbB 2, is located on chromosome 17, q21. It specifies a transmembrane receptor-like phosphoglycoprotein that is closely related to the EGFr (c-erbB-1). The ligand for c-erbB-2 is not known. A significant proportion of adenocarcinomas (especially of the breast, colon, and pancreas) have amplification and/or overexpression of c-erbB-2. The unique qualities associated with the subset of tumors that overexpress c-erbB-2 have not yet been firmly identified. PMID- 2565605 TI - P-glycoprotein and multidrug resistance in cancer chemotherapy. PMID- 2565606 TI - Comparison of methods for transcatheter fragmentation of gallstones. AB - Alternative methods have been considered for treating cholelithiasis. Compared to extracorporeal shockwave lithotripsy (ESWL), a percutaneous endoscopic approach would be more invasive, but would offer the advantage of immediate stone removal without the need for subsequent drug therapy. We performed an in vitro comparison of three methods of transcatheter cholecystolithotripsy with regard to effectiveness of stone fragmentation, damage to the gallbladder mucosa, and compatibility with percutaneous delivery systems. The three devices used for cholecystolithotripsy were the ultrasonic lithotriptor (UL), the electrohydraulic lithotriptor (EHL), and the thulium-holmium-chromium: YAG laser (THC:YAG). The UL effectively fragmented all types of stones studied, although it is necessary to hold the stone against the tip of the probe. The EHL quickly fragmented noncalcified and pigment stones simply by placing the tip in the vicinity of the stone, but calcified stones had to be held in position near the electrode. The THC:YAG was effective at fragmenting each type of stone, but the number of pulses required was quite large, corresponding to 7 min for some stones. The EHL had the most capacity for mucosal damage, followed by the THC:YAG laser. The UL produced no mucosal damage at the exposure times tested. The UL is not compatible with flexible endoscopes while the EHL and the THC:YAG are. Because of the specific advantages and disadvantages of each device, a combination of devices may be required for successful clinical cholecystolithotripsy. PMID- 2565607 TI - [Medical decision analysis in veterinary practice. An introduction with reference to the problem: to operate or not in cryptorchism in dogs]. AB - In the present report, medical decision analysis in veterinary practice is introduced. Fundamentals such as decision tree, utility, and sensitivity analysis are explained using a clinical example. The decision-analytic approach is found to produce an interesting result. Finally, a prospect for the future is presented. PMID- 2565608 TI - HLA-DR4 associated Dw types in rheumatoid arthritis. AB - Frequencies of HLA-DR4 and its related Dw types were compared between randomly selected normal controls and the index cases of multiplex rheumatoid arthritis (RA) families. A DR4 frequency of 68.3% was observed in index cases (n = 57) compared to 31.2% in normal controls (n = 96). Cellular typing with homozygous typing cells (HTCs) revealed significant increases of Dw4 (49.1% vs 22.9% RR = 3.2 p less than 0.001) and Dw14 (22.8% vs 2.1% RR = 13.9 p less than 0.001) in the index cases. A non-significant increase was seen for Dw13 (8.8% vs 4.1%). When DR4 positive patients and controls were compared, a significant increase was seen only for Dw14 (34.2% vs 6.6% RR = 7.3 p less than 0.01). Data from HLA genotyped RA and normal families allowed an examination of haplotype combinations of HLA-B antigens and DR4/Dw types to be made. HLA-Dw4 was predominantly found with B44 and Bw62 with nearly all DR4/Bw62 haplotypes being Dw4 positive. HLA Dw13 was associated with B44 and Dw14 with Bw60, B44 and B27. Based on HTC and normal family data. Dw10 was found to be strongly associated with B38 containing haplotypes. Analysis of 69 C4A, C4B complement typed DR4 haplotypes failed to show any statistically significant association between Dw type and "complotype". However, there was a suggestion of C4A3. BQO being associated with Dw4 (34.2% vs 16.1% X2 = 2.9 p = ns) and C4A3, B1 with Dw14 (45.5% vs 27.6% X2 = 2.1 p = ns).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565609 TI - Effects of prenatal methylmercury exposure on urinary proximal tubular enzyme excretion in neonatal rats. AB - The basal developmental pattern of excretion of 3 proximal tubular enzymes was determined in 8-h urinary specimens from neonatal rats. Gammaglutamyltransferase (GGT), alkaline phosphatase (ALP), and N-acetyl-beta-glucosaminidase (NAG) activities were measured at 3, 6, 9 and 12 days after birth. Subsequently, methylmercury chloride (CH3HgCl), known to induce foetotoxic changes in the proximal tubule was administered on days 8, 10 and 12 of gestation at 3 or 6 mg/kg and its effects on the enzyme activities were examined. Dose-related increases in the 3 enzyme activities occurred at dose levels that produced no maternal or postnatal toxicity, nor overt morphological malformation of the kidney. The peak of enzyme activities averaged about 200% and 130% of the control values for GGT, ALP, and NAG respectively, and occurred on days 3 and 6 in the treated groups. Urinary enzyme activities returned to the control levels from days 6 to 12. Our data point to the possibility of detecting CH3HgCl-induced prenatal effect on the kidney by measuring the 8-h urinary excretion of enzymes by rats in the early postnatal period. PMID- 2565610 TI - Studies on the induction of cholangiofibrosis by coumarin in the rat. AB - The histogenesis of coumarin-induced cholangiofibrosis in the rat has been determined. Proliferation of ductal structures was preceded by extensive damage to hepatocytes in the centrilobular region. Focal proliferation of ducts and fibrous tissue was present at 3 months and typical areas of cholangiofibrosis at 6 months. By 18 months the lesion was extensive and contained areas showing bizarre histological features suggestive of malignancy although no evidence of extra-hepatic metastasis was found. The lesion in animals returned to standard diet showed varying degrees of involution with extensive atrophy and fibrosis. A number of parameters of hepatic mixed function oxidase activity were reduced during the initial treatment period, at later times there was recovery of some microsomal enzyme activities. The activity of gamma-glutamyltransferase and the hepatic content of non-protein sulphydryl groups, in contrast, were raised throughout the treatment period. PMID- 2565611 TI - Diesel soot particles catalyze the production of oxy-radicals. AB - The formation of a strong oxidant similar to the OH. radical is catalyzed by diesel soot particles in the presence of cysteine and hydrogen peroxide or in the presence of light. The oxidant(s) formed causes fragmentation of methylthioketobutyric acid measurable as ethylene release. Furthermore, the model carotenoid crocin is bleached and thiobarbituric-acid-reactive material (malondialdehyde) is produced from linolenic acid. All reactions are inhibited by scavengers (propyl gallate, alpha-tocopherol, diazobicyclooctane) and by catalase. The reactions observed suggest that the toxicity and mutagenicity of diesel soot particles is at least in part due to the formation of reactive oxygen species. PMID- 2565612 TI - Biochemical studies in suicide victims: current findings and future implications. PMID- 2565613 TI - Does cyclosporin A affect neurotransmitters? Studies after kidney transplantation. PMID- 2565614 TI - RFLP analyses of naturally occurring intra-RT1 recombinant strains. PMID- 2565615 TI - Analysis of the major histocompatibility genes that control susceptibility to diabetes mellitus in the BB rat. PMID- 2565616 TI - Induction of suppressor T cells in vitro by an autologous alloreactive T cell line propagated from a DSBT-enhanced rat renal allograft. PMID- 2565617 TI - Laparoscopy and Fowler-Stephens orchiopexy in the management of the impalpable testis. AB - In children with an impalpable testis, laparoscopy is a valuable procedure that can be used in conjunction with inguinal and abdominal exploration to localize the testis. The instrumentation and technique of laparoscopy are described. If the testis is intra-abdominal, a Fowler-Stephens orchiopexy often is successful. Staging the Fowler-Stephens orchiopexy, with preliminary ligation of the testicular vessels in situ, followed 6 months later with transection of the vessels and orchiopexy, may improve the success of this technique by allowing collateral arterial flow to develop with minimal risk of arterial spasm. PMID- 2565618 TI - The human chromophobe cell renal carcinoma: its probable relation to intercalated cells of the collecting duct. AB - In the present study we have examined ten cases of the chromophobe type renal cell carcinoma. This type of tumor is distinguished from the other carcinomas of the kidney with light cytoplasm (formerly called "hypernephroid") by (a) a positive Hale's iron colloid stain of the cytoplasm, (b) the occurrence of numerous invaginated vesicles within the cytoplasm that resemble the invaginated vesicles of intercalated cells of the collecting duct system, and (c) a positive immunoreaction of both the plasma membrane and the cytoplasm with antibodies to the epithelial membrane antigen (EMA) and carbonic anhydrase C (CAC), respectively. Unlike oncocytomas, which also express CAC and EMA, the chromophobe renal cell carcinoma does not express the erythrocyte anion exchanger band 3. These findings strongly indicate that chromophobe renal cell carcinomas as well as oncocytomas of the kidney are histogenetically related to the two populations of intercalated cells of the collecting duct system. Thus, both tumors represent examples of renal tumors which disprove the broadly accepted hypothesis that all epithelial tumors of the kidney are histogenetically related to the proximal tubule. PMID- 2565619 TI - Acridine orange-mediated photodamage of microsomal- and lysosomal fractions. AB - Irradiation of microsomes with visible light in the presence of externally-added acridine orange results in O2 uptake, malondialdehyde accumulation, and inactivation of the microsomal drug-metabolizing system. The latter effect is reflected by a decrease in NADPH-cytochrome P450- and NADH-cytochrome b5 reductase activities and cytochromes P450 and b5 content by 88-, 85-, 60-, and 34%, respectively, after 5-min irradiation. Anoxia prevented inactivation of both reductases by 70-90%, whereas it prevented completely cytochrome b5 destruction. The presence of reducing equivalents, at the expense of NADPH and NADH, exert a partial protection (40-54% residual activities) against photosensitization damage on both reductase activities, whereas it almost fully protected cytochrome b5. Photosensitization of lipid peroxidation, as well as inactivation of the microsomal drug-metabolizing system, appears to involve both a type I and type II process. Products of lipid peroxidation might also play a role in enzyme inactivation and cytochrome destruction, as suggested by kinetic and time course studies and the redox state of microsomes. The uptake of acridine orange by isolated lysosomes is linearly dependent on the concentration of added dye and the distribution between extra- and intralysosomal acridine orange is strongly dependent on the amount of lysosomes. Irradiation of acridine orange-loaded lysosomes (light intensity at the sample position approximately 320 mW/cm2) produces an impairment of the membrane which leads to a rapid release of enzyme (N-acetyl-beta-glucosaminidase activity) into the medium, accompanied by a loss of activity in the lysosome-containing pellet and a partial photodamage of the enzyme. Concomitantly, thiobarbituric acid-reactive material accumulation increases in the reaction mixture with increasing irradiation time. When light intensity at the position was reduced to approximately 3.6 mW/cm2, photodamage of lysosomes was of a lesser magnitude, allowing the demonstration of a lag phase, which decreased with irradiation time, probably reflecting the so-called first stage activation of lysosomes, preceding the release of lysosomal enzymes. PMID- 2565620 TI - Complement activation in amyloid plaques in Alzheimer's dementia. AB - Amyloid plaques in Alzheimer's dementia contain complement factors C1q, C4 and C3. In the present study we demonstrate complement activation in amyloid plaques using immunoenzymatical techniques and specific antibodies against subunits of individual complement components and activated complement products. Amyloid plaques contain C1q and activated C3 fragments (C3c and C3d, g) but no C1s and C3a. These findings demonstrate that the complement components are not passively bound to the amyloid plaque structures but are the result of an activation process. The role of complement activation in the genesis of senile plaques is discussed. PMID- 2565621 TI - Evaluation of lysosomal stability in living cultured macrophages by cytofluorometry. Effect of silver lactate and hypotonic conditions. AB - The ability of living mouse peritoneal macrophages to retain the lysosomotropic photosensitizer acridine orange (AO) within their secondary lysosomes was studied with a novel cytofluorometric method. During exposure to blue light, cellular AO fluorescence turned from a red granular pattern to that of diffuse green. The resulting change in total fluorescence intensity versus time - a primary decline due to red fluorescence bleaching and a secondary recovery due to the spectral shift - was interpreted as the result of leakage of AO from the lysosomal vacuome. The hypothesis that this time course should be affected by changes in lysosomal membrane stability was tested by labilizing the lysosomes by exposure of cultured macrophages to either hypotonic medium or silver lactate. In hypotonic medium, the ability to retain AO decreased continuously. Exposure to low concentrations of silver lactate (10 microM) also decreased AO retention time. We suggest that this method could be used, within appropriate experimental conditions, to evaluate lysosomal membrane stability in living cells. PMID- 2565622 TI - Cytotoxicity of phenazine methosulphate on skeletal muscle. The role of the sarcoplasmic reticulum in initiating myofilament damage. AB - Phenazine methosulphate (PMS) or ferricyanide caused ultrastructural damage, including sarcolemma folds and swelling of the sarcoplasmic reticulum (SR), in amphibian skeletal muscle which corresponds with that triggered by a rise in [Ca]i and which, it is suggested, is caused by the activation of NAD(P)H oxidases at the sarcolemma (where it causes sarcolemma folding) and SR (where it causes myofilament damage). PMS also caused SR swelling and more limited damage in chemically-skinned muscle at zero [Ca]. In contrast with the oxygen paradox of cardiac muscle, there is no evidence for the production of oxygen radicals since no protection was provided by N2, mannitol, desferrioxamine or alpha-tocopherol, nor was the cell damage produced by an influx of Ca across the sarcolemma. PMID- 2565623 TI - Increase in B-cells in the pancreatic remnant after partial pancreatectomy in pigs. An immunocytochemical and functional study. AB - The regenerative and functional capacity of B-cells in the remaining pancreatic tissue after surgical removal of 40%, 60% and 80% of the pancreas was examined in 7 month old pigs (three animals in each group). Prior to resection and 1, 3 and 6 weeks after surgery, basal and glucose-stimulated levels of insulin and blood glucose were determined and compared with the preoperative data and that of sham operated controls. For quantitative morphology, the volume of the resected specimen and the residual pancreatic tissue, 6 weeks after surgery, was determined and sections evaluated by immunocytochemistry (insulin, glucagon, somatostatin, pancreatic polypeptide) combined with morphometry. In the remaining pancreas, the volume density of the B-cells was increased by 19% (1.57-1.92 after 60% resection; p less than 0.02) and 56% (1.57-2.38 after 80% resection; p less than 0.02) 6 weeks after surgery, compared with the respective resected portion of the pancreas and the controls (n = 12). The non-B-cells gained between 0-10% (PP-cells), 10-20% (D-cells) and 30-40% (A-cells) in the different resection groups. As the number of B-cells per given islet area remained unchanged (mean 4.12 cells/0.25 mm2), the increased volume density was due to an increase in cell number rather than cell size. Insulin secretion (integrated values, 0-120 min), was not significantly impaired after 40% and 60% resection (2711 +/- 250 all preoperative samples; 3215 +/- 474 40% at 6 week intravenous glucose tolerance test (IV-GTT); 1677 +/- 109 60% at 6 week IV-GTT), although the glucose levels (integrated values) were increased during the IV-GTT. The 80% resected animals showed a significant decrease in the insulin response only 1 week after surgery (integrated values: 2711 +/- 250 all preoperative samples, compared with 1250 +/- 508 1 week IV-GTT; p less than 0.05), while the integrated glucose values during IV-GTT (0-120 min) were significantly elevated throughout the observation period. These results suggest a B-cell hyperplasia in the residual pancreas after resection, which may cope with a normal functional demand, but disclose functional abnormalities when challenged with an increased glucose load. PMID- 2565624 TI - Localization of proinsulin and insulin in human insulinoma: preliminary immunohistochemical results. AB - We have carried out an immunohistochemical investigation of 15 human insulinomas applying monoclonal antibodies specifically recognizing proinsulin and insulin. Our results demonstrate that the epitopes unique to proinsulin and insulin can be detected with the respective monoclonal antibodies using the protein A-gold technique after routine formaldehyde fixation and paraffin embedding of the tissues. The immunostaining pattern for proinsulin and insulin in the insulinomas was different from the observed in B cells of pancreatic islets present in the adjacent normal pancreas. Furthermore, the pattern of immunostaining was found to vary from tumor to tumor. These findings strongly suggest the possibility of a disturbed proinsulin to insulin conversion in human insulinomas. PMID- 2565625 TI - Evidence of an acquired increase in mitogenesis in streptozotocin-diabetic rats, apparently relating to some tissue factor. AB - We have previously reported that rats which have been suffering from streptozotocin-diabetes for 4 weeks show a supranormal mast cell mediated mitogenesis in mesenteric windows and in the skin; this late emerging, augmented mitogenic responsiveness appears, to be unaffected by insulin per se. To test whether this increased proliferogenic response is effected by some acquired quality within the tissue rather than a systemic factor in the blood, we studied mast cell mediated mitogenesis in organ-cultured intact mesenteric windows from rats with diabetes of 4 weeks' duration, using a biochemically-defined serum-free growth medium. Mast cells were activated by Compound 48/80 and their secretion was quantified biochemically in terms of histamine release. The mast cell dependent mitogenic reaction in the predominant, morphologically discrete fibroblasts and mesothelial cells was quantified photometrically using Feulgen absorption analysis of individual cell nuclei, and by determination of the mitotic index. Both types of target cell responded to a significantly greater degree mitogenically in diabetic compared with control tissue. This finding suggests that a considerable part of the increased mitogenic responsiveness previously observed in diabetic animals in vivo is causally related to some tissue-bound, i.e., cellular and/or extracellular factor(s) acquired during the course of the disease. PMID- 2565626 TI - In situ hybridization--a useful tool for studies on collagen gene expression in cell culture as well as in normal and altered tissue. AB - We report the application of antisense RNA probes for in situ hybridization to identify collagen type I and type III mRNA synthesizing fibroblasts under in vitro and in vivo conditions in normal and wounded human skin. Non-specific hybridization was excluded by specific distribution patterns of alpha 1(I)- and alpha 1(III) probes in mouse fetuses. In addition, the specificity of hybridization was checked by sense probes, radioactively labelled transcripts of Gemini vectors and a keratin probe. In normal skin weakly activated fibroblasts were sparsely scattered within the dermis, while in wound healing processes mRNA both for alpha 1(I) and for alpha 1(III) was dramatically increased, thus suggesting that collagen synthesis is at least partly regulated at a pretranslational level. In addition, the intensity of the labelling, as defined by image analysis and the distribution pattern of collagen mRNA synthesizing cells, provide strong evidence that wound healing by primary intention starts within the deep dermis. PMID- 2565627 TI - Comparison of enzyme phenotypes in human bladder tumours and experimentally induced hyperplastic and neoplastic lesions of the rat urinary bladder. A combined histochemical and immunohistochemical approach. AB - The expression of a number of enzymes involved in drug metabolism, membrane function etc. was compared in hyperplastic and neoplastic lesions of the rat bladder and in human bladder tumours. Transitional cell carcinomas (TCC) in both rat and Man were characterized by decreased alkaline phosphatase (ALP) and increased gamma-glutamyl transpeptidase (GGT), beta-glucuronidase (beta-G1), succinate dehydrogenase (SD) and glucose-6-phosphate dehydrogenase (G6PD) activities. In addition, binding for antibodies specific for different cytochrome P-450 species (UT50, PB3a, MC1, MC2) and microsomal epoxide hydrolase (mEHb) was elevated in both murine and human tumours. Comparison of the enzyme phenotype in hyperplastic lesions induced by freeze ulceration or uracil administration with that in preneoplastic papillary or nodular hyperplasia (PNH) and TCC suggested, however, that most of the alteration in enzyme content or activity was non specific and related to requirements for epithelial cell proliferation. On the other hand, the decreased ALP, and increased GGT and beta-G1 activity appeared more directly related to neoplastic transformation. The results suggested that qualitative differences exist between reactive hyperplasia and preneoplastic or neoplastic lesions in the urinary bladder. The finding of increased cytochrome P 450, in clear contrast to the reduction characteristic of preneoplastic hepatic lesions, may be important with regard to the observed difference in neoplastic transformation between the bladder and liver in response to drug metabolising enzyme inducers. PMID- 2565628 TI - Compensatory adrenal growth in dexamethasone treated rats. AB - The changes in right adrenal weight and adrenocortical mitotic activity have been quantified in the early (up to 72 h) stages following left adrenalectomy or sham adrenalectomy in adult male Sprague Dawley rats. These have been compared with the changes seen in rats pretreated for 14 days with a daily intraperitoneal injection of the synthetic glucocorticoid, dexamethasone (200 micrograms/kg) body weight. The results indicate a significant proliferative response in both groups of animals, although basal proliferative activity and the amplitude of the response was lower in the dexamethasone treated animals. In addition, they suggest two waves of mitotic activity at 24 and 72 h. PMID- 2565629 TI - Further studies on the biological characteristics of an endogenous colon mitosis inhibitor: comparison with some structurally related peptides. AB - Previous work indicates that the colonic epithelial cell proliferation in mice is reversibly inhibited by the tripeptide pGlu-His-GlyOH found in aqueous extracts of the intestine. In the present study we examined the possible tissue specificity of the colon mitosis inhibitor. The mitotic rate in the small intestine, epidermis and forestomach in mice was registered after a single i.p. injection of the tripeptide. A significantly reduced rate of cell renewal was found at 18 h in the epidermis whereas no inhibition was observed in the forestomach or ileal epithelium. To investigate whether the amino acid sequence of the tripeptide is essential for the inhibitory effect, three structurally related bioactive peptides were tested and compared to the effect of CMI. CMI showed a bell-shaped dose-response relationship as previously shown, whereas the mitotic rate was not reduced in the colonic epithelium after treatment with either an epidermal mitosis inhibitory pentapeptide, or the dipeptide pGlu-GlyOH, or an analogue of luteinizing hormone-releasing hormone. The efficacy of the tripeptide was dependent on the basal rate of cell renewal in the colonic epithelium. When the tripeptide was given at the circadian nadir of cell proliferation a delayed reduction of proliferative activity was observed at 6 h after treatment, whereas treatment when the rate of cell proliferation was at its circadian zenith gave an immediate mitotic inhibition. PMID- 2565630 TI - Freeze-fracture study of plasma membranes in wild type and daunorubicin-resistant Ehrlich ascites tumor and P388 leukemia cells. AB - The plasma membrane is considered to play a major role in the development and maintenance of the multidrug resistance (MDR) phenotype, a role which may in part be mediated by an inducible 170 kD transmembrane protein (P-170). The present freeze-fracture study of plasma membranes of daunorubicin-resistant Ehrlich ascites and P388 leukemia cells demonstrated a significant increase in the density of intramembrane particles (IMP) in the P-face, but not the E-face, of resistant sublines compared with wild type cells. Furthermore, a three dimensional histogram plot of the diameters of P-face IMPs in Ehrlich ascites tumor cells showed the emergence of a subpopulation of 9 X 11 nm IMPs not found in wild type cells. The size of these IMPs would be consistent with a MW of approximately 340 kD, thus indicating that P-170, shown to be present in both resistant cell lines by Western blot analysis and immunohistochemical staining, exists as a dimer in the plasma membrane. Incubation with the calcium channel blocker verapamil, in concentrations known to inhibit daunorubicin efflux in resistant cells, showed evidence of membrane disturbance in the form of IMP clustering in both wild type and resistant Ehrlich ascites tumor cells. However, incubation with daunorubicin itself did not alter the freeze-fracture morphology of the plasma membranes. PMID- 2565632 TI - [Pharmacologic modification of mediator liberation and the effect of mediators]. AB - Blocking specific steps in the chain of events leading to disease is essential for elucidating pathogenesis. If antagonism to a given receptor does not modify the disease the receptor plays no role in the disease. In this review we describe specific points in the chain of events leading to respiratory disease at which pharmacologic intervention is possible and we describe results of such intervention. This includes interventions not presently available therapeutically. We distinguish three groups of interventions: interventions interfering with pre-receptor mechanism, those interacting with pharmacologic receptors or with proteins coupling receptor activation to cellular events and, finally, interventions affecting postreceptor mechanisms, e.g., second messenger systems including ion channels. Of the many interventions possible early pre receptor interventions are most desirable (preventive measures), but are often not available. Interventions at the receptor level, although available and in most cases specific for a given receptor are of limited effectiveness because multiple receptor involvement predominates in respiratory disease. Interventions at the post-receptor level have been most successful thus far presumably because post receptor mechanisms (e.g. second messenger systems) are shared by many receptors, therefore interventions at this level are largely independent of receptor(s) involved and have been most effective clinically. PMID- 2565631 TI - Expression of ras oncogene p21 during human fetal development as determined by monoclonal antibodies RAP-5, Y13-259, and DWP. AB - In this report we describe the expression of the ras proto-oncogene p21 protein in various tissues during normal fetal development. Conventional, formalin fixed and paraffin-embedded sections of normal organs were examined from fetuses ranging 9 to 42 weeks of gestation. Immunohistochemical localization of ras p21 was accomplished using the broadly reactive, mouse monoclonal antibodies RAP-5 and Y13-259. The monoclonal antibody DWP, which is specific for a mutated form of ras p21 having a valine/cysteine at amino acid position 12, was also used. Detectable expression of the p21 protein was seen at different time periods during fetal development depending on the tissue. The expression of ras p21 (as detected by RAP-5 and Y13-259) was noted in a wide range of cell types and tissues; intense immunostaining was noted in epithelial cells of the gastrointestinal tract, exocrine and endocrine pancreas, renal tubules and transitional urotheliem, as well as in other tissues. This immunostaining generally, but not invariably, corresponded with patterns previously reported in benign and/or malignant neoplasms of adult tissues. In most instances ras p21 expression, when present, occurred during periods of rapid growth in given organ systems. However, some actively proliferating fetal tissues such as thymus and spleen, failed to express detectable ras p21 suggesting that factors other than cell cycle may influence its expression. No reactivity with DWP was noted in any of the tissues, suggesting that the mutated forms detected by this monoclonal antibody are not expressed during normal human embryogenesis. These data show that there is regulated expression, and broad distribution of this gene product in normal developing human fetal tissue. PMID- 2565633 TI - [The development by I. P. Razenkov of the concept of neurohumoral regulation in the body]. AB - The problem of neurohumoral regulation is a key feature in the activities of I. P. Razenkov. I. P. Razenkov extended the notion of humoral regulation (1924-1937) and formulated the main aspects of the neurohumoral regulation concept. Further concretization of this concept was made on the model of the digestive system. The results of all studies have been generalized. PMID- 2565634 TI - Rapid isolation with Sep-Pak C18 cartridges and wide-bore capillary gas chromatography of some butyrophenones. AB - A simple and rapid method for isolation of five butyrophenones with Sep-Pak C18 cartridges from human samples, and their wide-bore capillary gas chromatography (GC), are presented. The GC was made by both flame ionization and electron capture detections. The drugs contained in alkaline samples were directly applied to the cartridges and eluted with chloroform/isopropanol (9:1). The recoveries with use of the cartridges were excellent for most drugs in both urine and plasma samples. We can recommend the Sep-Pak C18 cartridges for isolation of butyrophenones because of simplicity and rapidity, and also wide-bore capillary GC because of high sensitivity and low decomposition of drugs during passage through the column. PMID- 2565635 TI - [Physiological parameters of guinea pigs under long-term anesthesia with controlled respiration]. AB - 56 guinea pigs were anesthetized with a mixture of alpha-chloralose and a small dose of ethyl urethane injected i.p. in order to compare the chloralose-urethane anesthesia with neuroleptanesthesia, a mixture of droperidol-fentanyl and a small induction dose of Na-pentobarbital was injected i.p. to 14 animals. All animals were maintained at constant body temperature of 38 degrees C and artificially respirated via a tracheal cannula monitoring the end expiratory CO2 concentration. Mean arterial blood pressure (carotid artery) and heart rate were continuously recorded. Arterial acid-base status and pO2 were tested. Mean arterial blood pressure and heart rate were found to be somewhat higher in the neuroleptanesthetized animals. Under chloralose-urethane anesthesia and neuroleptanesthesia the mean arterial blood pressure amounted to 56 and 62 mm Hg, respectively. These findings correspond to data given in the literature concerning the mean pressure in unanesthetized guinea pigs. The mean heart rates of 295 and 316 min-1, respectively are somewhat higher compared to the normal range. Under both anesthesias, the arterial acid-base status was within the normal limits of unanesthetized animals. The mean arterial pO2 of 80.9 and 76.5 mm Hg was relatively low. The present studies have shown that chloralose-urethane as well as droperidol-fentanyl with pentobarbital are useful, under controlled artificial respiration, to achieve long-term anesthesia maintaining nearly normal systemic circulatory and respiratory conditions. PMID- 2565636 TI - gamma-Glutamyltranspeptidase in liver homogenates of rats of different ages: enzyme kinetics and age course of Km and Vmax. AB - Optimum incubation conditions for the determination of gamma glutamyltranspeptidase with liver homogenates from rats of different ages have been determined: about 1 mg protein/assay, 4.4 mM L-gamma-glutamyl-p nitroanilide, 40 mM glycylglycine, 9.6 mM MgCl2 in 0.1 M Tris-HCl buffer, pH 8.0; final volume 1.3 ml. The enzyme activity is very high at birth, the increase before birth is not continuous. After birth the enzyme activity decreases quickly over the first postnatal days. The ENDRENYI-KWONG-plot was linear with homogenates from newborn and 3-day-old animals. For all other age groups the graphs showed an angle. Statistical analysis carried out that a two-enzyme model plots the experimental data only insignificantly better than a one-enzyme model. It may be concluded that there are different GGT isozymes which show different affinities towards the substrate and which show different developmental patterns. PMID- 2565637 TI - [Neurohumoral and immune homeostasis in experimental toxoplasmosis in white rats]. AB - The intraperitoneal inoculation of Toxoplasma gondii strain RH into white rats is accompanied by the disorganization of choline-and histamine-energic activity, which, on one hand, facilitates the spread of the infective agent throughout the body and, on the other hand, activates nonspecific and specific protective factors. The toxoplasmic antigen appears in the blood stream as early as 24 hours after inoculation. Its penetration into tissues coincides with the activation of complement, this activation first following the alternative pathway and then, after the appearance of antibodies, the classical pathway. The increase of the titers of specific antibodies favors the formation of subcompensation in the acetylcholine and histamine systems and is accompanied by the elimination of the parasites from internal organs and their migration to the brain. Such complex immunological investigations provide useful information making it possible to explain some specific features of the pathogenesis of toxoplasmosis. PMID- 2565638 TI - [Adhesion in meningococcus]. AB - The adhesive activity of 113 meningococcal strains with different invasive properties and 10 Neisseria nonpathogenic strains have been studied. Adhesive properties have been revealed in 80-82% of these strains. Meningococcal strains isolated from the nasopharynx of carriers possess high hemagglutinating activity. The percentage of cells with pili in these strain was also higher than that in meningococcal strains isolated from the liquor of patients. This shows the presence of direct correlation between the number of pili in the cells (according to the data of electron microscopy) and their activity in the hemagglutination test, which makes it possible to use this test for the determination of the adhesive capacity of meningococci, associated with the presence of pili. PMID- 2565639 TI - [Successful autotransplantation of intra-abdominal testes into the scrotum using microvascular technics]. AB - The surgical treatment of undescended and nonpalpable testes still remains controversial. The shorter the vascular stem, the greater difficulties are during orchidopexy. The results of the formerly used "stage" and "Loop" techniques are unsure and frequently lead to postoperative atrophy of the testes. Thanks to the marked development of the microsurgical technique, over the past 10 years, besides the conventional methods for orchidopexy of the testes with a short vascular stem, microvascular revascularization on the autotransplanted testes from the abdomen into the scrotum is also used. During the last year two boys were successfully operated upon with this method with which this microvascular anastomosis technique is described and the problems openly discussed. PMID- 2565640 TI - Definition of CD 11a, b, c, and CD 18 glycoproteins on chemotactically deficient granulocyte membranes in patients affected by myeloid disorders. AB - We studied neutrophil chemotaxis and surface membrane glycoproteins in 12 patients suffering from myeloid disorders with abnormal karyotype using in vitro techniques in all 12. Chromosome studies were also carried out virtually simultaneously. We chose to study only patients showing a deficit of chemotaxis (p less than 0.001). Ten of these patients revealed a clonal chromosome abnormality in most of the leukemic cells. The monoclonal antibody technique using MoAb 60.1, 60.3, 60.5, OKM1, LFA-1 and 9E8 demonstrated normal leukocyte membrane glycoproteins CD 11a, b, c and CD 18 and therefore excludes that an abnormality of these is involved in the pathogenesis of altered leukocyte chemotaxis. No correlation between chemotactic deficit and any specific clonal chromosome abnormality was found. PMID- 2565641 TI - Molecular characterization of Italian chromosomes carrying the Lepore Boston gene. AB - Hemoglobin Lepore Boston is characterized by abnormal non-alpha-chains that are the product of a fusion delta beta gene originated from an unequal crossing over between misaligned delta and beta genes. The investigation of the restriction fragment length polymorphisms (RFLP) of the beta-globin gene cluster in 18 Italian Lepore Boston chromosomes indicates that the hybrid gene is linked to two RFLP patterns. The majority of chromosomes show a pattern which corresponds to haplotype V and a minority to haplotype I according to Orkin's classification. A single Hb Lepore Boston homozygote was homozygous for haplotype V. The two haplotypes differ only for a single site 3' to the beta cluster. Our data allow the speculation that in Italy the Lepore Boston gene might be the result of multiple recombination events. PMID- 2565642 TI - Electron microscopical localization of guanylate cyclase activity in the neocortex of the guinea pig. AB - The localization of the guanylate cyclase (GC) activity has been established in the neocortex of adult guinea pigs by means of electron microscopical histochemistry [the DMSO-method of Fujimoto et al. (1981)]. Reaction product was deposited within a population of large- and medium-sized cortical neurons as well as in the cytoplasm of a part of the dendrites of variable size and in the cytoplasm and the nuclear membrane of a number of protoplasmic astrocytes. In the perikarya of the positive neurons, the reaction precipitate was mainly located within the cisterns of the rough endoplasmic reticulum and on the nuclear membrane. In the dendrites, the reaction product was usually distributed in close contact with microtubules, microfilaments, and beneath the postsynaptic membranes of a number of axodendritic synaptic contacts. The axons and all presynaptic boutons were negative. Thus, the localization of the GC could be determined as exclusively postsynaptic. The results obtained support the view for the probable participation of cyclic GMP in the cholinergic, glutaminergic or GABAergic, or peptidergic transmitter mechanisms in the central nervous system. PMID- 2565643 TI - Application of immunogold-silver staining and immunoenzymatic methods in multiple labelling of human pancreatic Langerhans islet cells. AB - The use of immunogold-silver staining (IGSS) combined with immunoperoxidase and/or immunoalkaline phosphatase methods for the simultaneous demonstration of pancreatic islet cell hormones on routinely fixed paraffin-embedded human tissue sections was examined. If IGSS was applied first, the black colour of silver enhanced colloidal gold on doubly immunostained sections contrasted with the colours of most of the chromogens used generally in the 2 immunoenzymatic methods. If IGSS was followed by immunoalkaline phosphatase and immunoperoxidase techniques in optional sequence, 3 different hormone-containing cell types could be stained simultaneously without non-specific cross-reactions. IGSS and immunoalkaline phosphatase methods, together with 2 kinds of non-cross-reacting immunoperoxidase systems, permitted the detection of 4 distinct antigens on the same tissue section. Multiple immunohistochemical labelling of the endocrine pancreas provides an opportunity for the correct and rapid analysis of the topographic and morphometric relationships between different hormone-producing cell populations under both normal and pathological conditions. IGSS is of great potential for the simultaneous immunolabelling of antigens situated within separate cells. PMID- 2565644 TI - Early stages of late onset Alzheimer's disease. Diagnostic criteria, protein metabolism, precursor loading effects, neurochemical and neuropsychological applications. PMID- 2565645 TI - Astrocytic pathology of methionine sulfoximine-induced encephalopathy. AB - To investigate the roles imposed on astrocytes for glutamate metabolism, a specific inhibitor of glutamine synthetase (GS), methionine sulfoximine (MSO), was repeatedly administered to rats and histopathological changes were correlated with glycogen accumulation and the immunocytochemistry of GS and glial fibrillary acidic protein (GFAP). Prolonged MSO-loading (every 12 h up to three times, 100 150 mg/kg body weight) brought about the appearance of astrocytes with swollen, watery nuclei reminiscent of Alzheimer II glia chiefly in the neocortex, hippocampus and lateral thalamus after 24 h. Concomitantly, profound accumulation of glycogen ensued in the superficial three layers of the neocortex, hippocampus and pyriform cortex. GS immunoreactivity appeared enhanced in the cortex, hippocampus and lateral thalamus with parallel increase in GFAP immunoreactivity after prolonged treatment. Oligodendrocytes in the diencephalon and brain stem also normally contained GS immunoreactivity. Some animals developed necrotic lesions in the dorsolateral neocortex. The area of glycogen accumulation coincided with the known distribution of N-methyl D-aspartate (NMDA) glutamate receptors and, thus, GS may play important roles in NMDA receptor-mediated glutamate metabolism. The Alzheimer II type changes, however, did not correlate with NMDA-receptor distribution. These results indicate certain regionalizations in the roles of astrocytes and oligodendrocytes in glutamate and ammonia metabolisms. PMID- 2565646 TI - Excretory function after unilateral renal denervation and administration of propranolol to unanaesthetized dogs. AB - The experiments were carried out on female dogs with exteriorized ureters prior to and following surgical denervation of the left kidney. Propranolol 1.0 mg/kg b.w. was administered intravenously. Sodium, potassium, chloride, calcium, magnesium, zinc, copper, creatinine and urea concentrations in the urine from the denervated and intact kidneys as well as in blood drawn were determined. After renal denervation PAH clearance was determined. As a result of denervation diuresis and calcium and copper excretion were increased while urine osmolality was diminished. No change occurred in kidney blood flow and GFR. After propranolol administration diuresis, calcium and copper excretion in the intact kidney significantly increased. Changes in the excretory function of the left kidney following its denervation were not a result of alterations in renal haemodynamics. Results obtained indicative of that beta-adrenergic receptors contribute to the excretion of calcium and copper ions. PMID- 2565647 TI - [Immediate complications of radical cystectomy. Intestinal fistula: our experience in the treatment with somatostatin]. AB - During the period of time between November 1986 and May 1988, 14 total cystectomies with one-step urinary derivations were performed in our Service. The surgical techniques utilized include: 7 Bricker, 2 Camey, 1 ileocecal segment derivation, 2 cutaneous transureterostomies and 2 cutaneous ureterostomies. Among the complications observed there were 3 intestinal fistulas which were treated at first in a conservative way with parenteral nutrition and somatostatin. An immediate response was obtained in all three cases with a decrease in the fistula deficit. Surgical treatment was required in two patients after a lack of response to conservative treatment. The fistula closed in the third patient, to open again spontaneously 15 days later. It was controlled with conservative means and closed again 45 days later. PMID- 2565648 TI - [The advantages and disadvantages of surgical prevention of thrombosis]. AB - The need for providing for a prophylaxis against thrombosis in surgery is now generally acknowledged in view of a basic risk of 10-60% thromboses and 1-5% fatal pulmonary embolisms. Initial efforts to recognise risk of thrombosis in patients on the basis of certain characteristic signs have not been successful, so that general prophylaxis is now fundamentally preferred. Measures of physical prophylaxis of thrombosis have been only partly successful, but the effort required to apply them in practice is out of all proportion to the effect achieved, so that this type of prophylaxis can only be used as a complement to drug therapy and with special high-risk patients. Although anticoagulation is effective, its rate of side effects is so high and the technical difficulties involved are so great that this method cannot be recommended in general prophylaxis of venous thrombosis. This also applies, albeit less dramatically, to thrombose prophylaxis with dextranes; in fact, these must be used with caution only, especially in elderly patients. Completely useless but characterised by considerable side effects is the attempt to achieve venous thrombosis prophylaxis via aggregation inhibitors. Today the standard method in surgery is low-dose heparin prophylaxis according to the effectivity-risk calculation. By applying the conventional non-fractionated heparins according to the low-dose schema it was possible to lower the pulmonary embolism mortality in all surgical disciplines with the exception of traumatology and orthopaedics to one third and the thrombosis rate to one third and the thrombosis rate to one third to one fourth of the original risk.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565649 TI - [Differentiated therapy of distal radius fracture]. AB - The fracture of the distal radius loco typico is one of the most frequently occurring fractures. This article presents types of fractures, in particular Colles', Smith's, Barton's and multiple-fragment's fractures. Conservative and operative treatment is described, as well as problems related to the different procedures. Besides fracture retention by plaster splint or cast, Besides fracture retention by plaster splint or cast, transcutaneous osteosynthesis by Kirschner wire, lag screws, osteosynthesis by buttress plate and the use of the external fixator are described as examples of operative therapeutics. PMID- 2565650 TI - [Thoracic trauma in the pre-hospital phase]. AB - Preclinically relevant injuries of the thorax are described patho physiologically. The diagnosis at the scene of accident and the treatment of the emergency of the functional after-effect injury are described. Respiratory insufficiency requires early artificial respiration. A pneumothorax should be drained, at least, on the respired patient. If there is a pneumothorax associated with tension due to the confined air, the relief would be obligatory. The relief should take place after the digital opening of the thorax by silicon drainage. If there is a mediastinal emphysema with a seriously haemodynamic effect, the relief would be indicated by collar mediastinostomy. If there is a pericardium tamponade with circulatory collapse after a perforating trauma, the patient must be brought to the medical attention of a surgeon as quickly as possible. The delay in time must not be the consequence of the relief of puncture. If a person injured in an accident who has a rupture of the trachea can not be respired immediate exposure of the rupture site is imperative. Resuscitation measures in case of an injured thorax must be done at the opened thorax. PMID- 2565651 TI - [Statistical evaluation of 4,141 osteosyntheses with autocompression plates]. AB - Statistical display of the cases of our hospital who underwent osteosynthesis with autocompression plate between 1968 and 1986. Based on the unselected evaluation of our surgical reports and computer monitoring an analysis of the total of 4.141 ACP-osteosynthesis is given according to the various origin types of surgical procedures, age and sex distribution and chiefly complications of the operative revisions. The total of revised complications was 4.18%, among that the rate of infections was 1.37%, of pseudarthroses 0.92%, breaking out of plates 2.2 per thousand and of material fractures 1.9 per thousand. From these data a high security of the osteosynthesis by autocompression plates concluded. PMID- 2565652 TI - [Observations on post-traumatic osteitis]. AB - The following study reports the frequency, the promoting factors, the therapy and her results of the 32 patients with posttraumatic osteitis we treated between 1984 and 1986 in the Surgical University Clinic Tubingen. Most of the primary injuries were caused by traffic-accidents; especially dangerous were those with motor-bikes, which led frequently via open fractures of the shank to osteitis. We saw the highest infection-rates after plate-osteosyntheses. The infects became obvious in most cases either about one month after the accident or a year later coinciding with the increased use of the limb. The most frequent bacterium was Staph. aureus both in the mono-infections and in the mixed-infections forming a third of the group. We always performed an operative therapy with the intention to stabilize the bone and to clean the infection-site. The strict performance of this management led to infect-suppression in all cases with the need of only one amputation. PMID- 2565653 TI - [Displacement of the tibial tuberosity in retropatellar cartilage damage and recurrent patellar dislocation]. AB - In our hospital in the years 1980 till 1986 37 displacements of the tuberositas tibiae were performed in chondropathia patellae and recurrent luxation of the patella. The follow-up examination took into consideration the radiological and clinical findings, the subjective complaints and the ability of going in for sports. These clinical experiences and pressure measurements in the femoro patellar joint show the best results after the displacement of the tuberositas tibiae according to Blauth. Additionally we recommend an accurate indication for chondropathia patellae and recurrent luxation of the patella and accompanying surgical measures. PMID- 2565654 TI - [Dynamic transfixation in unstable dislocation fracture of the proximal interphalangeal joint]. AB - Unstable fracture dislocations of the PIP-joint are difficult to manage, because the primary joint destruction and the necessity of immobilisation often result in joint stiffness and the loss of immobilisation may promote chronic subluxation. In the case described in the article we treated the unstable fracture dislocation with a dynamic transfixation with a "force couple splint" following the open reconstruction and internal fixation of the destroyed joint. The application of this dynamic transfixation allowed early active mobility exercising of the PIP joint during bone healing and produced an excellent result. PMID- 2565655 TI - [Surgical treatment of fresh subcutaneous rupture of the Achilles tendon and results of its treatment]. AB - In the years between 1981 and 1986 32 patients with a subcutaneous rupture of the Achilles tendon were operated on. Two different surgical techniques were preferred: using the tendon of the m. plantaris as suture according to Streli, and Bunnell's suture. In a clinical exploration 17 patients could be recorded. According to the classification of Trillat, 70.6% of the patients show a very good result. That means that the patients have the same sports and professional efficiency as before the injury and operation, a painless Achilles tendon, a stable heel and unrestricted mobility. PMID- 2565656 TI - [Traumatic dislocations/dislocation fractures of the cervical spine. Neurosurgical experiences in 42 surgical patients]. AB - Neurosurgical results in 42 patients operated on for cervical spine dislocations or fracture-dislocations are described. The study shows that repositioning with the Crutchfield extension is almost always successful. Imbricated fractures must occasionally be repositioned surgically (2 cases). A strikingly large proportion of patients (50%) in whom decompression was carried out ventrally were found to be suffering from traumatic disc hernia in the lower cervical spine. This was often accompanied by laceration of the posterior longitudinal ligament. Operative stabilization of unstable fractures, especially pseudarthrosis, of the odontoid process was performed in the main by dorsal immobilization (blocking) at C1/C2 (pelvic bone chips/wiring), whereby the residual rotational limitation was only of moderate degree and well tolerated subjectively. Two patients in whom stabilization was achieved in this way developed secondary ossification of the old pseudarthrosis of the odontoid process. In 2 cases the operation was carried out by the transbuccal route, again with the use of pelvic bone chips. Operative stabilization below C2 was performed 15 times using the ventral approach only, namely 13 times by Cloward's method and twice by Caspar's method. A combined ventrodorsal approach was used a total of 13 times in patients with severely damaged dorsal ligaments; here we made use ventrally of autologous pelvic bone chips, dorsally of wire osteosynthesis. The results were in the main good in respect of repositioning, uniformly good in respect of stability. In the great majority of cases with medullary or radiculomedullary deficits, the operation resulted in complete healing or at least an improvement, namely in 75% of patients with lesions of the odontoid process and in 81% of those with lesions below C2. In the second part of the study, the advantages and disadvantages of the various methods are set out and discussed in the light of recent advances and in particular of operative experience with tumours in the region of the cervical spine. PMID- 2565657 TI - [Conservative fracture treatment of the cervical spine with halo fixation]. AB - A differentiated therapeutic approach is necessary in discoligamentous or osseous lesions of the cervical spine. Whereas lesions with neurological disturbances are stabilised by operation almost in all cases--and extensive lesions of the lower cervical spine (C3 to C7) quite frequently--, conservative treatment is the order of the day in sequels to trauma at the upper cervical spine (C1 and C2) as well as in mild lesions of all sections of the cervical vertebral column. Fractures of the dens of Types II and III (after Anderson and d'Alonzo) are the exception, since they are now being stabilized by surgery on a large scale. The advantages of the halo-fixateur therapy compared against extension and immobilization in Minerva gypsum are that secondary correction of positioning is possible; that functionally disturbing and extended spondylodeses are avoided; that care of the polytraumatized patient is facilitated; that X-ray films are easy to assess; and that the period of hospitalization is greatly reduced. Successful treatment is possible only of the possible complications are known. Failures are possible if the head screws become loose, if there are pin-trac infections, if dislocations and fractures are redislocated, and if there are points of pressure beneath the jacket. PMID- 2565658 TI - [Stability of the spine following osteosynthesis with internal fixation. Case report]. AB - Fractures of the first and third lumbar vertebrae were primarily reduced and stabilized with a Fixateur interne (The XII-LIV) and filled with transpedicular bone grafts in a 21 years old girl. Nine months after this injury she fell from the fifth floor sustaining a flexion-distraction fracture of Th XI whereas the lumbar spine as well as the Fixateur interne remained intact. Stabilization by a Fixateur interne was carried out again. This is an example of the broad application of Fixateur interne in lower thoracic and lumbar spine traumatology and for the high degree of stability of the Fixateur system. PMID- 2565659 TI - [Modified transpeduncular spongiosa-plasty with osteotomy of the transverse process]. AB - Bone grafting of the vertebral body is an essential part in stabilisation of the spine. A modified transpeduncular approach to the vertebral body is described. Using this approach a bone graft can be placed more exactly in a shorter time. PMID- 2565660 TI - [Spongiosa formation in plate osteosynthesis--a comparative animal experiment study of current and auto-compression plates using the Zespol principle]. AB - Osteosynthesis with modern self-contracting plates has a few fundamental drawbacks apart from its undisputably good points. Among these drawbacks we have the pressure strain on the bone surface; the traction strain exercised by the screw thread in the bone; and, chiefly, spongiotisation and bone atrophy due to so-called stress protection. A new modification of this process, called Zespol, in which the plate is no longer fastened direct to the bone but is fastened floating over the bone surface to so-called platform screws, tries to solve these problems. In a comparative animal experiment tibial transverse osteotomies in the rabbit were treated by osteosynthesis using small fragmentary ACP and Zespol plates. In respect of healing of the fracture none of the two methods showed any advantage. Histological evaluation of the plated bone sections were characterised in both plate systems by increasing spongiotisation and bone atrophy that increased with the length of application. Degradation processes were about equal with both systems. A special feature with the Zespol animals was that the cleft originally existing between the plate and the bone was filled up with newly formed spongy and later cortical bone. Due to this process the original cortex layer lost its function and atrophied severely. After 12 to 18 weeks the bone surface had reached the lower side of the Zespol plate and there was contact by touch. This restored similar biomechanical conditions as those existing in conventional plates after this period. PMID- 2565661 TI - [Percutaneous Goetze cerclage in torsion fractures of the tibia. A computer assisted follow-up of 186 cases]. AB - 186 lower leg torsional fractures treated via percutaneous cerclage wiring (Gotze's method) yielded good to very good final results. There were no intraoperative complications such as lesions of the vessels and nerves. The number of refractures (1.61%), the incidence of delayed healing (4.3%), infections (3.23%) and misalignments (6.9%) are all comparable with data that can be found in the literature. No pseudoarthroses were seen. Besides low rate of complications and the rapidity of the procedure, the simplicity and low cost of the required instrumentarium are also very attractive. The fact that the patients remain in the hospital for a short while only, is yet another contribution to keeping the cost of general health care low in these times of exploding expenditure for medical attendance. PMID- 2565662 TI - [Rotational deformities following pediatric femoral shaft fracture]. AB - At the beginning of 1983 65 children who had been treated at our hospital for fractures of the femoral shaft could be followed-up both clinically and radiologically at an average of 29 months after the injury. The radiological examination was performed with special regard to the rotational deformity; the torsion was measured using Rippstein's method. Out of 65 patients, 19 patients showed a difference between the angles of antetorsion of more than 10 degrees (29.2%) which we considered to be "rotational malpositions" (8 patients with endorotational and 11 patients with exorotational deformities). 40% of all fractures localized in the proximal third of the femur were accompanied by a rotational deformity of more than 10 degrees as compared to the other side. Exactly 4 years later a second examination of antetorsion was made using the same methodical procedure. 18 out of the 19 patients with rotational malpositions and 2 patients with a difference between the AT-angles of exactly 10 degrees could be re-examined. The average correction in the overall population was 3.4 degrees ( 10 degrees to +5 degrees). In 15 cases we found a decrease in the difference between the angles of antetorsion by 5.8 degrees on an average, 1 patient showed no change in the difference, and in 4 patients the difference had increased by 3 degrees on an average. In 7 of the 20 patients (35%) the difference between the angles of antetorsion was less than 10 degrees, whereas the rotational malposition persisted in 13 cases. The extent of correction was much larger in patients with exorotational deformities than in those with endorotational deformities. A very close relation could be seen between the age of the children and the extent of correction. Furthermore, rotational deformities caused by fractures of the proximal third of the femoral shaft showed significantly less tendency towards correction than fractures with a more distal location. PMID- 2565663 TI - [The "positioned image" of the talo-calcaneus joint in instability of the posterior distal ankle joint]. AB - With regard to the very common disease of sprained ankles the combined tear with ruptures of ligaments of the subtalar joint are possible. Even an isolated instability is known as the reason of long lasting post-traumatic complaints. Using the apparatus of Scheuba, well-known in diagnosing instabilities of the knee and ankle joint, a new standardized method controlling instabilities of the subtalar talo-calcanear joint is introduced. In an experimental investigation this radiological technique has been tested out in cadaveric ankle joints (nine joints) with artificial ligamentous lesions. The results show that in all cases an instability is recognizable radiologically using this technique. For clinical use the bilateral examination is recommended failing standardized values for unilateral use. PMID- 2565664 TI - Addictive behaviors and benzodiazepines: 1. Abuse liability and physical dependence. AB - A review of scientific and clinical evidence indicates: (1) benzodiazepines have an abuse liability and can cause physical dependence, (2) abuse liability is of a lower order of magnitude than that associated with common intoxicants such as barbiturates, opioids or stimulants, (3) sustained, exclusive use of benzodiazepines for inducing intoxication occurs but, it is infrequent, (4) benzodiazepines tend to be secondary drugs to a preferred primary intoxicant; in experimental paradigms normals prefer placebo to benzodiazepines, (5) susceptibility to physical dependence varies widely as low doses are sufficient to produce it in some but very high multiples are not sufficient to produce it in many others, (6) factors predisposing to physical dependence are: total lifetime dose, previous exposure to drugs cross-tolerant, such as alcohol or barbiturates, concomitant severe medical/psychiatric problems, and severe persisting stress. Individual susceptibility to abuse and to become dependent on benzodiazepines should be investigated much more vigorously than it has heretofore. PMID- 2565665 TI - Muscle strength, activity, housing and the risk of falls in elderly people. AB - In a national survey in Britain, 983 elderly people randomly selected from eight areas were asked about falls they had had in the past. The independent association of social and physical variables with a history of one or more falls was analysed by multiple logistic regression. Those who had fallen one or more times had reduced grip strength and were less mobile than those who had not fallen. More of them used non-phenothiazine tranquilizers, lived alone, had recently lost weight or were physically disabled. Independent of these influences, a history of having fallen was strongly related to place of residence. Part of this relationship was explained by differences in housing, in particular the percentage of houses without indoor lavatories. PMID- 2565666 TI - Guinea pig alveolar macrophages possess beta-adrenergic receptors. PMID- 2565667 TI - Analysis of the beta-adrenoceptor mediated inhibition of IgG1 and IgE dependent guinea-pig anaphylactic tracheal smooth muscle contraction. PMID- 2565668 TI - Effect of azelastine on microviscosity of bronchial lavage fluid obtained from actively sensitized and challenged guinea pigs. AB - Fluorescence polarisation technique was adapted to measure microviscosity in the bronchoalveolar lavage fluid (BAL) of guinea pigs. In sensitized guinea pigs, 20 hours after antigen challenge, the microviscosity of BAL was increased by 86%, suggesting that antigen-induced bronchospasm is followed by inflammatory events. Pretreatment with azelastine (3 mg/kg, p.o., 2 hours prior to antigen challenge) tended to normalize microviscosity in the bronchoalveolar lavage fluid obtained from challenged guinea pigs. The present results suggest that azelastine inhibits the increase of microviscosity of BAL, a symptom of the late phase reactions. PMID- 2565669 TI - Effects of cooling on in vitro responses of human peripheral airways to inflammatory mediators and neurotransmitters. AB - In asthma, inhalation of cold dry air induces bronchoconstriction. It has been suggested that cooling of the airway wall might induce this bronchoconstriction. Therefore the effects of cooling on the contractility of human peripheral airways were studied in vitro before and during lowering of the temperature. Cooling relaxed human airways in vitro and reduced responsiveness to methacholine, histamine and LTC4 both in terms of -logEC50 and maximal effect. It also reduced the maximal effect of isoprenaline. There was no measurable alpha-adrenergic activity before or during cooling. We conclude that cooling does not sensitize human airways to any of these agonists and that cold-induced bronchoconstriction in vivo is not due to a direct effect of cooling on airway smooth muscle. PMID- 2565670 TI - Antigenic cross-reactions between tick-borne orbiviruses of the Kemerovo serogroup. AB - Antigenic relationships between the four subgroups of the Kemerovo serogroup of tick-borne orbiviruses were examined. Kemerovo (KEM subgroup), Broadhaven (BRD) and Wexford (WEX) [Great Island (GI) sub-group], Chenuda (CNU sub-group), and Wad Medani (WM sub-group) viruses cross-reacted in immunofluorescence tests. Complement fixation tests (CFT) revealed that KEM virus was more closely related to BRD and WEX viruses than to either CNU or WM viruses. By cross-neutralization, all the viruses were shown to be distinct; only BRD and WEX showed slight cross reaction. In Vero cells infected with either KEM, BRD, WEX, or CNU viruses, 10 major (p1 to p10) and a variable number of minor virus-induced 35S-labelled polypeptides were detected. Comparison of the polypeptides precipitated by homologous AF revealed close similarities between BRD and WEX viruses but obvious differences between these two viruses and KEM and CNU viruses. In cross reactions, p6 of KEM and CNU viruses, and p7 of BRD and WEX viruses were immunoprecipitated. Immunofluorescence and immunoprecipitation tests appeared to identify an inter-group specific antigen that was distinct from an intra-group specific antigen detected by CFT. The results support division of KEM-related viruses into 3 new serogroups - KEM (comprising KEM and GI subgroups), CNU and WM - and indicate that a 38 to 43kD polypeptide carries the major inter-group specific antigen. PMID- 2565672 TI - Rapid diagnosis of adult diarrhea rotavirus (ADRV): detection of viral antigens in faecal samples using staphylococcal co-agglutination test. AB - Staphylococcus aureus Cowan I rich in protein A when sensitized with guinea pig antiserum to adult diarrhea rotavirus (ADRV) at 1:16 gave a strong co agglutination with ADRV-positive faecal samples as previously confirmed by electron microscopy (EM) and enzyme-linked immunosorbent assay (ELISA). The bacteria sensitized with normal guinea pig serum did not give any co agglutination. Blocking tests using rabbit ADRV-specific antiserum for the treatment of twelve ADRV-positive samples abolished the reaction. All the fifty ELISA-confirmed ADRV-positive faecal samples gave positive co-agglutination, whereas all the forty-eight ELISA-negative faecal samples from healthy subjects gave negative results. The test has been proved to be rapid, simple, specific, and economic, useful for rapid diagnosis even in remote areas, so that the ADRV infection can definitely be differentiated from some of acute bacterial diarrheas. PMID- 2565671 TI - A rapid method for detection of flavivirus antigens: staphylococcal co agglutination test using monoclonal antibodies to Japanese encephalitis virus. AB - Staphylococcus aureus rich in protein A when coated with monoclonal antibodies (MoAb) to Japanese encephalitis virus (JEV) gave a highly specific reaction with flavivirus antigens. The bacteria coated with JEV species-specific MoAb gave a strong co-agglutination with fifty-six JEV isolates from various parts of China, but no co-agglutination with Murray Valley encephalitis (MVE) and Kunjin (Kun) virus antigens. The flavivirus- and subgroup-specific MoAbs were reactive with MVE and Kun, as well as with the majority of the JEV strains. Blocking test with homologous MoAbs abolished co-agglutination further confirming its specificity. Numerous virus particles were observed on the surface of MoAb-coated staphylococci under the electron microscope after co-agglutination. The test appeared rapid, specific, simple to perform, and useful for rapid detection and identification of flaviviruses. PMID- 2565673 TI - Molecular and biological properties of a variant of avian influenza A/Seal/Massachusetts/1/80 (H7N7) virus that is pathogenic for mice. AB - A/Seal/Mass/80 influenza virus has been shown to be closely related antigenically and genetically to avian influenza H7N7 viruses, however, the virus does not replicate efficiently in avian species but does replicate in most mammals, except mice (Hinshaw et al., Infect. Immun., 34, 351-361, 1981). In order to develop a model defining the molecular changes that occur during acquisition of virulence, the A/Seal/Mass/80 virus was adapted to growth in mouse lungs. The adaptation was accompanied by changes in a number of properties of the haemagglutinin as well as by changes in other genes of the virus as determined by RNA: RNA hybridization. PMID- 2565675 TI - Amplification of arbovirus transmission by mosquito intradermal probing and interrupted feeding. AB - Probing is the crucial phase for the successful intake of the blood by a mosquito female, saliva being ejected during the intradermal probing period. When Tahyna virus (California group, family Bunyaviridae) carrying and transmitting Aedes aegypti mosquito was allowed to feed on 3 suckling white mice for 4 hr, 66.7% of the 162 exposed mice became infected. When one infected mosquito was put in contact with 5 mice, 53.6% of the 250 exposed mice became infected. Multiple transmission of the virus to the available hosts during completion of one blood meal by a single mosquito has been demonstrated. PMID- 2565674 TI - Occurrence and aetiology of acute respiratory diseases: results of a longterm surveillance programme. AB - Totals of 58,661,000 acute respiratory disease (ARD) cases, 1,376,651 bronchitis and pneumonia complications, and 93,042 deaths from influenza, bronchitis, pneumonia or chronic pulmonary affection were notified during 11 years of ARD surveillance from 1975 to 1986. All ARD seasons started with the first phase in September-December; this increase in morbidity was caused chiefly by adenoviruses, parainfluenza viruses, rhinoviruses and M. pneumoniae. Second wave of ARD morbidity occurring in January-April used to be explosive and was associated with an influenza epidemic in 9 of the 11 seasons; only in 1978/79 and 1984/85 the ARD epidemics were caused by adenoviruses and especially RSV, the share of influenza being minimal. Pneumonia and bronchitis excesses occured during epidemics caused by M. pneumoniae in 1975/76, 1980/81 and 1985/86. Particularly high mortality excesses occurred in 1976, 1977 and 1983 during epidemics elicited by a new drift variants of influenza A(H3N2). Identification of viral agent of M. pneumoniae attempted in 5474 ARD cases was successful at 37.4%. The respective contributions of parainfluenza viruses, adenoviruses, influenza A virus and RSV to overall aetiologically identified morbidity were 14.2, 13.9, 13.8, and 12.0%. Mixed infections (2-3 agents identified simultaneously) accounted for 14.6% of cases. Type B influenza virus, rhinoviruses, enteroviruses and herpes simplex virus contributed only by 5.6 7.8%. In ordinary seasons the share of M. pneumoniae in aetiologically identified ARD morbidity was 0.6-3.8%; this agent displayed predominance at 5-year cycles, when accounting for 20.5-38.9% of cases. The most frequently detected agents in individual age groups were as follows: in preschool children parainfluenza (18.6%), RSV (16.6%), and adenoviruses (17.4%); in school children M. pneumoniae (26%), influenza A and B (10.2 and 14.7% respectively), and adenoviruses (10.7%); in adolescents and young adults influenza type A (20.2%), M. pneumoniae (15.0%), and rhinoviruses (13.3%); in adults above 25 years age influenza A virus (38%), and other respiratory viruses at a frequency lower than 10% each. PMID- 2565676 TI - Effects of pH and ionic strength on precipitation of phytopathogenic viruses by polyethylene glycol. AB - The effects of ionic strength of the solution (changed by varying NaCl concentrations or buffer molarity) on the precipitation with polyethylene glycol (PEG) 6000 were studied on phytopathogenic viruses of different morphology: the isometric red clover mottle virus (RCMV), rod-shaped tobacco mosaic virus, flexuous potato virus X (PVX) and bacilliform alfalfa mosaic virus. With increasing NaCl concentration or buffer molarity up to a certain level (0.1 mol/l), the efficiency of PEG precipitation increased. This relationship did not apply to PVX. The effects of pH on PEG precipitation were studied on RCMV. The efficiency of precipitation increased with decreasing difference between pH of the solution and pI of the virus. PMID- 2565677 TI - Chemical composition of phase I Coxiella burnetii soluble antigen prepared by trichloroacetic acid extraction. AB - Optimal conditions of extraction (time and temperature) by trichloroacetic acid of soluble antigen from phase I Coxiella burnetii (TCAE), possessing protective properties and used as a chemovaccine against Q fever in men, were studied. Extracts prepared under various conditions were analysed for their polysaccharide, protein and phosphorus contents. Forty-five min of extraction at 0 degrees C were sufficient to obtain a soluble antigen reacting in immunodiffusion with hyperimmune rabbit antiserum. The polysaccharide contents decreased with prolonged extraction at 0 degrees C. At higher extraction temperatures (37 and 100 degrees C), the polysaccharide contents increased while that of proteins decreased. TCAE prepared at 100 degrees C gave no positive immunodiffusion reaction. PMID- 2565678 TI - Immunoblotting analysis of class-specific antibody response in patients with primary HSV infection. AB - Class-specific immune response in acute herpes simplex virus (HSV) infections to individual virus specified polypeptide antigens was analysed by immunoblotting. The HSV specified glycoproteins B (gB), C (gC), and D (gD) were detected. IgG antibody response was shown to develop to various other virus specific polypeptides as well. The IgM- and IgA-antibody responses remained restricted to only a few HSV specific proteins, namely VP 13 (80 kD), VP 20 (52 kD), and VP 23 (40 kD) and to the low molecular weight polypeptides. PMID- 2565679 TI - Wesselsbron virus infection in West African dwarf goats (Fouta djallon): virological and immunological studies. AB - West African dwarf goats were experimentally infected with Nigerian strain of Wesselsbron virus. Viraemia was detected in infected goats 2 days after infection and lasted for one day. A 100% mortality was observed among the infected animals; the virus was reisolated in mice from almost every tissue obtained from the bodies of infected goats. In addition, the infected goats developed complement fixing and haemagglutination inhibiting antibodies to Wesselsbron virus. PMID- 2565680 TI - Total haemolytic complement profile in chicks following fowl pox vaccination or infection. AB - Total haemolytic complement levels were assessed in normal, fowl pox-vaccinated or infected chicks using radial immune haemolysis up to 28 days post-treatment. Significantly lower values of total haemolytic complement were recorded 7-21 days post-vaccination or 21 days post-infection as compared to controls (p less than 0.05). The differences between intervals, the influence of the period of treatment were also significant, but the vaccinated chicks did not differ significantly from the infected ones (p less than 0.05). It is concluded that lower circulating levels of total haemolytic complement may be due to deposition of complement at the sites of virus replication. PMID- 2565681 TI - Serologic survey of wild rodents in Georgia for antibodies to orthopoxviruses. PMID- 2565682 TI - The effect of beta blockade on single photon emission computed tomographic (SPECT) thallium-201 images in patients with coronary disease. AB - We evaluated the effect of beta blockers on thallium-201 (Tl-201) single photon emission computed tomographic (SPECT) imaging in 12 patients with coronary disease using an automated computer algorithm. Maximal exercise heart rate and blood pressure were reduced and exercise time was increased with beta blockers. Estimated stress defect size decreased from 47 +/- 36.3 gm during placebo treatment to 32 +/- 27.1 gm during beta blocker therapy (-32%; p less than 0.01). The placebo treatment redistribution defect was estimated to be 28 +/- 29.8 gm. It fell to 15 +/- 23.3 gm with beta blockade (-46%; p less than 0.005). All patients had a stress Tl-201 defect during placebo treatment and eight had redistribution defects consistent with residual scar. During beta blocker therapy, 2 of 12 patients had normal stress-redistribution studies and only five patients had redistribution defects. Beta blockade can reduce exercise and redistribution Tl-201 SPECT defect size significantly while simultaneously increasing exercise time and reducing angina. Beta blockers may unmask or may eliminate evidence of redistribution. Tl-201 SPECT imaging may be useful in defining the reduction in ischemia produced by cardiac drugs. PMID- 2565683 TI - The mechanisms of exercise provocation of supraventricular tachycardia. AB - Treadmill exercise tests, electrophysiologic studies, and isoproterenol infusions were performed in 14 patients with exercise provocable supraventricular tachycardia to delineate the mechanisms of exercise provocation of paroxysmal supraventricular tachycardia. Treadmill exercise tests reproducibly provoked supraventricular tachycardia in all patients. Supraventricular tachycardia similar to that provoked by exercise occurred spontaneously during isoproterenol infusions in 9 of 11 patients tested. The specific supraventricular tachycardia diagnoses of all patients were atrial reentrant tachycardia (two patients), automatic atrial tachycardia (three), atrial flutter-fibrillation (one), atypical junctional tachycardia (two), and orthodromic atrioventricular (AV) reentrant tachycardia (six) as defined by electrophysiologic studies. Various mechanisms of exercise or isoproterenol induction of supraventricular tachycardia were identified. A critical heart rate and/or appropriate sympathetic state was found to provoke all instances of reentrant or automatic atrial tachycardia and atypical junctional tachycardia. A properly timed atrial premature beat provoked five of six cases of AV reentrant tachycardia and the only case of atrial flutter fibrillation. The remaining case of AV reentrant tachycardia was induced by a ventricular premature beat. In conclusion, the mechanisms of exercise provocation of reentrant or automatic supraventricular tachycardia are multiple and include a critical sinus rate, increased sympathetic tone, and properly timed atrial or ventricular premature beats. PMID- 2565685 TI - Stability of esmolol hydrochloride and sodium nitroprusside in intravenous admixtures. AB - The stability of esmolol hydrochloride and sodium nitroprusside in an admixture containing both drugs was studied. Solutions containing sodium nitroprusside in a final concentration of approximately 200 micrograms/mL and esmolol hydrochloride in a final concentration of 10 mg/mL in 5% dextrose injection were prepared in a 250-mL volumetric flask. The flask was wrapped with a light-protective cover, stored at ambient room temperature (15-30 degrees C), and protected from light. All experiments were conducted in triplicate with samples taken at 0, 2, 4, 8, and 24 hours. Testing included measurement of pH and absorbance at 400 and 600 nm. High-performance liquid chromatography was used to measure esmolol hydrochloride and sodium nitroprusside concentrations. No changes were observed in the physical appearance, pH, or absorbance of the admixtures. Neither the esmolol hydrochloride nor the sodium nitroprusside concentrations varied by more than 4% during the study. Under the conditions studied, esmolol hydrochloride is compatible with sodium nitroprusside in an admixture containing both drugs. PMID- 2565684 TI - One-year prognosis of primary ventricular fibrillation complicating acute myocardial infarction. The GISSI (Gruppo Italiano per lo Studio della Streptochinasi nell'Infarto miocardico) investigators. AB - The 1-year prognosis of 293 patients discharged alive from the hospital after acute myocardial infarction (AMI), who experienced primary ventricular fibrillation (VF) in the acute phase, was compared with that of a reference group of 6,337 patients identified from the same population included in the Gruppo Italiano per lo Studio della Streptochinasi nell'Infarto miocardico (GISSI) trial. There was no difference in the 6- and 12-month mortality between the patients with primary VF and the reference group (3.7 vs 2.7% and 4.1 vs 4.2%, respectively). Survival of the 2 groups was also similar when patients were stratified according to infarct site (anterior and posterior), and whether or not they received treatment with streptokinase during AMI. Thus, long-term mortality of patients discharged alive after AMI complicated by primary VF is low and is not influenced by previous fibrinolytic therapy or by infarct site. The excess mortality of patients with primary VF is confined to the hospital phase, after which survivors represent a low-risk subgroup. PMID- 2565686 TI - Visual compatibility of intravenous famotidine with selected drugs. PMID- 2565687 TI - Isradipine in essential hypertension: the Belgian General Practitioners' Study. AB - Over 200 hypertensive patients were recruited by 37 general practitioners into a single-blind 12-week study to assess the efficacy, tolerability, and safety of isradipine as an antihypertensive, alone and in combination with guanfacine. A total of 212 patients were given isradipine at doses of 1.25 and 2.5 mg twice daily. Twelve hours after the last dose, diastolic blood pressure was reduced to no more than 90 mm Hg in 52.6 percent of patients treated with isradipine alone. After eight weeks of treatment, 30 percent of patients were also given guanfacine 1 mg daily. By Week 12, 67.6 percent of the patients had attained normotension. Compared with placebo, side-effect frequency was higher for flushing and edema with isradipine, and dry mouth was more frequent with added guanfacine. Electrocardiographic examinations and routine laboratory determinations showed no clinically relevant changes. These data indicate that isradipine as monotherapy and in combination with guanfacine is an effective antihypertensive agent. Most patients will continue to participate in a two-year follow-up involving bimonthly clinical visits and half-yearly electrocardiographic examinations and laboratory determinations. PMID- 2565689 TI - Safety and efficacy of isradipine, alone and in combination, in the treatment of angina pectoris. AB - Five hundred ninety outpatients aged 18 years or older with stable angina pectoris entered a multicenter, single-blind, nonrandomized, baseline-controlled study to assess the efficacy, safety, and tolerability of isradipine in doses of 2.5, 5, or 7.5 mg three times daily for 12 weeks, following a two-week placebo "washout" period. Patients were assessed at the initial visit and, thereafter, every two weeks with a final evaluation at Week 14. The final mean dose was 5.9 mg three times daily. Overall, isradipine was found to reduce significantly the angina attack rate and nitroglycerin consumption in patients with chronic, stable, effort-induced angina pectoris. Isradipine was generally well tolerated when prescribed alone or with concomitant beta-blocker medication. PMID- 2565688 TI - First-line and combination treatment for hypertension. AB - Conventional stepped-care approaches for the control of high blood pressure based on the use of diuretics and beta-blockers have had a disappointing impact on coronary heart disease, the major cause of death in hypertension. Drug therapy for blood pressure should be complementary to attempts to control other risk factors and should not adversely affect other factors. A range of drugs, including calcium antagonists, angiotensin-converting enzyme inhibitors, and alpha-blockers, can be considered as possible first-line treatment. The choice of drug therapy should be individualized and the minimal number of drugs used in the simplest rational regimen. Simple demographic factors and associated diseases, risk factors, and activities should be used to select the most appropriate agents. New combinations offer alternatives for patients who do not receive adequate control with monotherapy. PMID- 2565690 TI - Progression to AIDS in patients with lymphadenopathy or AIDS-related complex: reappraisal of risk and predictive factors. AB - PURPOSE: In 1985, we reported that acquired immunodeficiency syndrome (AIDS) developed in 14 of 81 (17%) men with generalized lymphadenopathy followed prospectively for an average of 13 months. The presence of oral thrush or constitutional symptoms, or both, or severely impaired T4+ cell responses to specific antigen (interferon-gamma production) accurately identified patients at immediate risk for AIDS. The purpose of the current report is to describe the progress of these 81 patients during the three and a half years since enrollment and to include new data on initial serum levels of beta 2 microglobulin and human immunodeficiency virus (HIV) p24 antigen. PATIENTS AND METHODS: The mean age of the 81 patients was 35.4 years; 79 were homosexuals and two were drug abusers. Immunologic testing was performed once at the time of enrollment in all patients. Seventy-seven of the 81 patients were seropositive for HIV antibody. Frozen samples of serum, also obtained at initial study, were assayed in 1988 for beta 2 microglobulin and HIV p24 antigen. The clinical status of patients was determined six, 14, and 36 months after enrollment was closed (June 1984) by either interview and examination or telephone contact with private physicians. RESULTS: After three and a half years of follow-up, 42 patients have developed AIDS, including (1) 77% who had had thrush or symptoms, or both, (2) 80% to 88% of those who originally demonstrated marked immunologic abnormalities (skin test anergy, less than 200 T4+ cells/mm3, T4/T8 cell ratio of less than 0.5, severely impaired interferon-gamma production [less than 25 U/mL], or elevated serum beta 2 microglobulin level [greater than 3.0 mg/L], and (3) 95% of patients with HIV p24 antigenemia. However, AIDS also developed in 51% of patients who had had more apparently benign initial manifestations (lymphadenopathy alone, herpes zoster), in 41% to 54% despite normal initial results for either T4+ cell number, interferon-gamma secretion, beta 2 microglobulin, or skin testing, and in 44% of those whose sera did not contain HIV antigen. CONCLUSION: These updated results demonstrate the remarkably poor prognosis of patients with generalized lymphadenopathy or AIDS-related complex irrespective of initial clinical, immunologic, and serologic findings, and suggest that essentially all such persons may be candidates for antiviral therapy. PMID- 2565691 TI - Alcoholics' use of benzodiazepines. PMID- 2565692 TI - Immunohistochemical approach to the functional morphology of the hypothalamic hypophysial system. AB - Immunohistochemical studies at the light and electron microscopic levels have provided much information on functional morphology in the hypothalamic hypophysial system. The present paper describes the immunohistochemical techniques available at present and their use to determine the localizations of neurons containing hypophysiotrophic substances, the co-storage of plural signals in these neurons, and the synaptic regulation of these neurons in rats. PMID- 2565693 TI - [Mechanomyography and electromyography--2 competing methods of relaxometry using vecuronium]. AB - The recording of the evoked twitch tension has been the established reference method to quantitate neuromuscular blockade. The evoked compound electromyogram has been introduced later as a clinically more convenient alternative. We compared both methods in 20 patients in whom cumulative dose response curves of vecuronium and the time constants of weaning neuromuscular blockade were determined. The evoked twitch tension was slightly more sensitive in detecting residual neuromuscular blockade than the EMG. No significant differences between the two methods were found in the cumulative 90% blocking dose, the duration of block and the recovery time (25%-75%). These results are in agreement with communications of previous authors using different nondepolarizing muscle relaxants. In the absence of abnormal conditions such as neuromuscular disorders and hypothermia, recording of evoked electromyography is a clinically satisfactory method to quantitate neuromuscular blockade. PMID- 2565694 TI - An assay for dihydroorotase using high-performance liquid chromatography with radioactivity detection. AB - An assay for measuring dihydroorotase activity was devised. Radiolabeled substrate and product were separated by high-performance liquid chromatography using a reverse-phase column with ion-pairing, and the radioactivity was quantitated by flow detection. PMID- 2565695 TI - Characterization of benzodiazepine drugs by ion mobility spectrometry. AB - Chemical ionization ion mobility spectrometry (CI-IMS) was used to characterize a number of benzodiazepines. In almost every example studied, the positive ion mobility spectrum consisted of a single ion peak corresponding to [M]+ or [MH]+. With some compounds, e.g., oxazepam, lorazepam, and chlordiazepoxide, fragment ions were noted that serve as good markers for the identification of these chemicals. Reduced mobility constants (K0) for the most significant peaks were calculated, and all ions produced were mass-analyzed by injection into a quadrupole mass spectrometer. The results of this study point to the potential of IMS as a qualitative tool for the rapid detection (analysis time less than 10 s) and reliable identification of benzodiazepines. Preliminary results on the application of digital signal processing and a second-derivative algorithm to partially overlapping IMS peaks are presented, and potential improvements are discussed. PMID- 2565696 TI - Co-localization pattern of growth hormone (GH) and prolactin (PRL) within the anterior pituitary cells in the female rat and female musk shrew. AB - By a double immunocytochemical labeling procedure, using the protein A-gold method combined with electron microscopy, the co-localization pattern of growth hormone (GH) and prolactin (PRL) was detected in the anterior pituitary cells of female rats and female musk shrews. Two types of co-localization of GH and PRL were demonstrated. First, GH- and PRL-containing secretory granules were intermixed within closely aggregated and interdigitated cell clusters that were composed of GH and PRL cells. This phenomenon was characteristically seen in pregnant rats and pregnant musk shrews. Therefore, the occurrence of an intermixture of GH and PRL granules might be related to an enhanced cellular function for PRL synthesis. In another pattern of co-localization of GH and PRL, both hormones were co-packaged in the same secretory granules within a single cell. Such cells were scarce, small, irregularly shaped, and observed only in pregnant rats. These mixed GH-PRL cells contained not only mixed GH-PRL granules but also granules containing only GH or PRL. This suggests that these bihormonal cells are able to synthesize, synchronously or asynchronously, GH and PRL. Furthermore, granule extrusion from the mixed cells was clearly shown in this study. It seems likely that the mixed GH-PRL cells reveal active cellular function in the pituitary gland of the pregnant rat. PMID- 2565697 TI - [Comparison in healthy volunteers of the systemic beta adrenergic receptor blockade by beta blockader eye drops]. AB - The isoproterenol dose-response curve was used to assess quantitatively the degree of systemic beta-adrenoceptor blockade induced by metipranolol (Betanol) and betaxolol (Betoptic) eye drops. The study was carried out in twelve healthy volunteers, aged 22 +/- 1.4 yr. In a randomized double-blind trial, each volunteer received, on separate occasions at least one week apart, one drop in each eye of either placebo (physiological saline) or either of the ophthalmic beta-blockers. The intraocular pressure (Pio), heart rate (fc), arterial systolic (Pasys) and diastolic (Padia) pressures were measured before instillation of the eye drops after 15 to 30 min rest, and 3 h afterwards. The isoproterenol dose response curve was studied 3 h after instillation of the drops. The CD25 (the amount of isoproterenol needed to increase fc by 25 b.min-1) was obtained by extrapolation on the least square linear regression curve. Both beta-blockers gave a significant fall in Pio compared with placebo, metipranolol more than betaxolol (p less than 0.02). There was also a significantly greater fall in fc with both metipranolol and betaxolol than with placebo. There were no changes in Pasys and Padia. CD25 was significantly increased with both beta-blocker eye drops as compared with placebo (p less than 0.05 for betaxolol; p less than 0.01 for metipranolol), but there was no difference between the two. Systemic absorption after topical application of ocular beta-blockers was thus confirmed for both metipranolol and betaxolol. However, the degree of beta-adrenoceptor blockade was weaker than that observed with other older ocular beta-blockers (timolol and carteolol).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565698 TI - [2 cases of pulmonary excision in patients with acquired immunodeficiency syndrome. What is the role of surgery in AIDS?]. AB - The authors report two cases of lung resection in patients with AIDS: left upper lobectomy for diagnosis and treatment of an active, localized lung lesion which proved to be tuberculosis and atypical resection of the right apex and pleurodesis for an infected pneumohydrothorax. The postoperative course was uneventful despite the fragile clinical context. The place of surgical indications for diagnostic purposes is reduced by the good results obtained with endoscopic investigations and limited to failures of these procedures and to pulmonary sarcomatous lesions. The place of therapeutic surgical indications is essentially reserved for pleural complications (pleurodesis, pleurectomy, pleurotomy). Surgery may be unjustified in tracheostomised patients at the stage of respiratory failure. It may have a small but effective role with reduced risk in active, localized lung lesions. The surgery must be performed according to simple and classical by very rigorous rules of protection for the operators and the environment. PMID- 2565699 TI - N-methyl-D-aspartate antagonists: ready for clinical trial in brain ischemia? AB - Antagonists of the N-methyl-D-aspartate (NMDA) subclass of glutamate receptors may offer a new approach for the treatment of ischemic brain injury. This strategy is supported by a well-developed scientific foundation and encouraging results in a variety of in vivo and in vitro experimental models. Several specific antagonists, including MK-801, dextrorphan, dextromethorphan, and ketamine, have already been used at low doses in humans for other indications and are potential candidates for Phase I clinical trials. PMID- 2565700 TI - Role of G proteins in the regulation of the cardiovascular system. AB - A precise description of the involvement of G proteins in regulation of the cardiovascular system is not possible at the present time although it is clear that they do have important regulatory roles. The cardiovascular system is composed of a variety of cell types, which are subject to control by several different hormones, as well as by hormones that have several different effects in the same cell type. Although, historically, variations in the type and number of receptors located on each cell have been used to explain this diversity of hormonal responses, we must now consider the large number and diversity of G proteins in any effort to understand the coordinated hormonal regulation of cellular functions. Given that there are eight known G proteins and several others have been speculated, each of which is composed of three subunits, each of which has several different forms, the possible combinations of subunits into functionally distinct G proteins is enormous. To place this newly described family of G proteins into the appropriate hormone signaling pathways will require a continued research effort. However, with recent progress in producing specific antibodies to each of the G protein subunits, it may now be possible to determine the specific receptor-effector functions of each G protein and their individual subunits. PMID- 2565701 TI - Carbohydrate-specific monoclonal antibodies bind to human granulocytes and stimulate antibody-dependent cellular cytotoxicity. AB - Mouse monoclonal antibodies which specifically recognize human granulocytes are used to study the classification, differentiation, and function of these cells. Mouse monoclonal antibody WEM-G1 specifically binds to human neutrophils and eosinophils. It also affects granulocyte function by stimulating granulocyte mediated antibody-dependent cellular cytotoxicity. Biochemical studies presented here show that WEM-G1 recognizes the sugar sequence 3-fucosyllactosamine, Gal beta 1-4[Fuc alpha 1-3]GlcNAc. This sequence is present in granulocyte glycolipids and in glycoproteins of average approximate Mr 165,000 and 105,000. WEM-G1 is thus similar to other monoclonal antibodies that recognize this sequence on granulocytes and various other cells. Some of these 3 fucosyllactosamine-specific antibodies affect several other granulocyte functions. Knowledge of the biochemical structure of the WEM-G1 antigen suggested testing granulocyte function with other monoclonal antibodies of similar specificity. Antibodies recognizing both the identical oligosaccharide structure and a related sequence, Gal beta 1-4GlcNAc-R, were also found to stimulate granulocyte-mediated antibody-dependent cellular cytotoxicity. PMID- 2565702 TI - The recombinant spinach acyl-acyl carrier protein-I expressed in Escherichia coli is the 18:1 delta 11(cis) thioester. AB - A synthetic spinach acyl carrier protein-I (ACP-I) gene was cloned and expressed in the Escherichia coli beta-alanine auxotroph SJ16 (P. D. Beremand et al. (1987) Arch. Biochem. Biophys. 256, 90-100). After characterization of the transformed cells and purification of the protein product it was evident that 50% of the recombinant spinach ACP-I was acylated during early log-phase growth (D. J. Guerra et al. (1988) J. Biol. Chem. 263, 4386-4391). We have purified the recombinant acyl-acyl carrier protein-I to greater than 90% homogeneity and have made a fatty acid methyl ester of the delipidated and trypsin-treated preparation. We have found that the acyl moiety attached to recombinant spinach acyl carrier protein-I is 18:1 delta 11(cis) (cis-vaccenic acid) a major unsaturated end product of Escherichia coli de novo fatty acid synthesis. This result reflects previous work (D. S. Guerra et al. (1986) Plant Physiol. 82, 448 453) which suggested the acyl carrier protein-I structure has evolved from ancestral ACP structures to accommodate the eukaryotic pathway of lipid synthesis in higher plants. The accumulation of recombinant 18:1 delta 11(cis) acyl carrier protein-I in transformed E. coli SJ16 cells attests to the poor reactivity of this substrate to acyl transferase reactions and may help explain the lack of effect on pools of fatty acids found in vivo. PMID- 2565703 TI - [Recovery kinetics of hematopoiesis after peripheral blood stem cell autotransplantation]. AB - Hematopoietic recovery kinetics was evaluated in five children with therapy refractory cancers who received peripheral blood stem cell autotransplantation (PBSCT) following marrow-ablative chemotherapy. Four children received graft containing more than 1.5 X 10(4) CFU-GM/kg and days required to achieve granulocyte counts of greater than 0.5 X 10(9)/l were 6, 10, 13, and 18, respectively. One patient received 0.9 X 10(4) CFU-GM/kg, but, recovery of granulopoiesis was retarded (greater than 30 days). The days required to achieve platelet count of greater than 50 X 10(9)/l were 12, 15, 16, 195, and greater than 240, respectively. Transient decrease of blood cell counts developed 3 to 5 weeks after transplantation and thereafter, the recovery of hematopoiesis became stable. With the use of a large number of progenitors, PBSCT seems to be safe and effective new type of stem cell rescue operation as an alternative to bone marrow transplantation. PMID- 2565704 TI - [Sialyl Lewis(x)-I (SLX) as a tumor-associated carbohydrate antigen in sera in patients with gastric and colorectal cancer--evaluation according to clinico pathological factors]. AB - Sialyl Lewis(x)-i (SLX) as a new tumor-associated antigen was assayed in sera of 133 gastric cancer and 62 colorectal cancer patients with preoperative state in order to elucidate the clinico-pathological features. The percent positives in sera of patients with gastric and colorectal cancer were 12.7% (17/133) and 16.1% (10/62), respectively, and there were few positive cases in the early stages. Therefore, it is suggested that the early diagnosis for gastric and colorectal cancer using SLX alone may be difficult. However, it is revealed that SLX has a high tumor-specificity in consideration of the clinico-pathological properties such as H, P, n, ly, v factors, depth of cancer invasion and operative radicality. These data suggested that SLX could be applicable as a useful tumor marker which shows the metastasis or extension of carcinoma. PMID- 2565705 TI - Immunosuppression in men with bowenoid papulosis. AB - The immune status of four men with bowenoid papulosis was evaluated. Each case had been refractory to multiple methods of treatment. Three of the men had other infections and demonstrated a depletion of T4-helper cells. Two of these patients were anergic on skin testing, and the third showed weak reactivity. The fourth patient, who had no evidence of additional infections, had a normal T4 value and T4/T8 ratio, but was anergic on skin testing. All the men were serologically negative for human immunodeficiency virus antibodies. One of the immunosuppressed patients developed squamous cell carcinoma of the tongue, which, along with his bowenoid papulosis, contained human papillomavirus 16 DNA. We suggest that patients with persistent bowenoid papulosis be investigated for altered immune status and followed up as potential candidates for the development of epithelial malignant neoplasms. PMID- 2565706 TI - Identifying akinesia and akathisia: the relationship between patient's self report and nurse's assessment. AB - This study examines the relationship between the self-reports of 96 out-patients at four community mental health centers and nursing assessments of these same patients regarding drug-induced akinesia and akathisia. Akinesia, diminished spontaneity characterized by motor slowness and stiffness, and akathisia, a feeling of inner restlessness, are common but often misdiagnosed side effects of neuroleptics. Significant positive correlation was found between self-reports and nursing assessments. Data indicate that specific subjective responses as well as systematic clinical examination should be included when assessing for drug induced akinesia and akathisia. PMID- 2565707 TI - HLA class II DR, DQ, and DP restriction fragment length polymorphisms in rheumatoid arthritis. AB - HLA class II restriction fragment length polymorphisms (RFLPs) were studied in 43 individuals with established seropositive rheumatoid arthritis (RA) and in a group of healthy controls. All patients and controls were tissue typed for HLA-A, B, and DR antigens. Rapid, initial screening for RA associated RFLPs was conducted by pooling DNA samples from 11 HLA-DR4 positive patients with RA and comparing the RFLP patterns with those seen in a pool of DNA samples drawn from 11 HLA-DR4 positive healthy controls. Candidate RA associated RFLPs were examined in our full panel of patients with RA and controls. In most cases the RFLPs detected showed no significant association with RA. An exception was a 13.0 kb DraI DQ beta associated RFLP, which, when HLA-DR4 positive patients with RA and controls were considered alone, showed a weak positive association with susceptibility to RA. This RFLP was not associated with known DR, DQ, or Dw specificities. These results show a distinct paucity of class II RA associated RFLPs but may indicate a role for DQ beta genetic variation in the aetiology of RA. PMID- 2565709 TI - Reduced level of sex-specific antigen (H-Y antigen) on lymphocytes in some patients with bilateral cryptorchidism. AB - Sex-specific (Sxs) antigen on the surface of nucleated cells from normal human males seems to be essential for the formation of testes. The relative quantity of the antigen on lymphocytes was evaluated by absorption experiments in a complement-dependent cytotoxicity test or in an ELISA technique using antisera against Sxs antigen produced by immunization of female rats. Lymphocytes from 13 normal males were Sxs-antigen positive, and cells from 12 normal females were characterized as Sxs-antigen negative. However, in the testing of lymphocytes from nine boys with bilateral cryptorchidism, only six revealed a normal male absorption pattern, whereas the antigen level on cells from three boys, all of them with normal karyotype, was reduced compared with the normal male level. No correlation between Sxs-antigen level and testosterone response after treatment with hCG could be demonstrated. PMID- 2565708 TI - Extrathoracic and transthoracic management of vascular disease of the aortic arch branches: a 16-year experience. AB - To determine factors influencing results of operation for proximal brachiocephalic arterial disease, a 16-year review of 74 consecutive patients was undertaken. A total of 62 subclavian, 14 common carotid, 6 innominate, and 2 vertebral arteries were revascularized during 79 procedures. The approach was transthoracic in 12 operations and extrathoracic in 67. Mean follow-up was 57 +/- 45 months. Two patients (2.5%) died of stroke after extrathoracic revascularization of the common carotid artery. Cumulative 5-year and 10-year freedom from neurological events was 81% and 75%, respectively. The best results were obtained with transthoracic procedures, with revascularization of the subclavian artery rather than the common carotid artery, and in operations performed in patients without associated distal carotid disease. In view of the recent progress in operative techniques and postoperative surgical care, the choice between the transthoracic approach and the extrathoracic approach should not be biased; rather, they should remain equal and viable alternatives based on anatomical and clinical features of the individual patients. PMID- 2565710 TI - Neuroleptic responsiveness in siblings concordant for schizophrenia. PMID- 2565711 TI - Duodenal somatostatinoma with psammoma bodies. AB - A duodenal somatostatinoma was found incidentally in a 60-year-old woman undergoing cholecystectomy. Microscopically, the tumor had a glandular architecture with abundant psammoma bodies. Electron microscopy revealed tumor cells resembling D-cells of the pancreatic islets. Immunohistochemically, there was staining for neuron-specific enolase, chromogranin, and diffuse cytoplasmic positivity for somatostatin only. By immunoelectron microscopy, somatostatin was identified predominantly in lucent membrane-bound secretory granules. X-ray dispersive microanalysis showed the psammoma bodies contained calcium apatite crystals. This case is compared with other reported cases with a description of cellular localization of somatostatin and development of psammomatous calcification. PMID- 2565712 TI - [Endocrine cells of the APUD-system in the human lung (electron microscopy characteristics)]. AB - Lungs of 4 human fetuses (11-, 13-, 22-, 28-week-old), of 1 stillborn and of 3 mature persons, operated in connection with pulmonary cancer, have been investigated. In the fetal lungs apudocytes and neuroepithelial bodies (NEB) have been revealed. The apudocytes differ from each other by structure and size of endocrine granules. In the 11-week-old fetus P1 cells with two types of granules occur most often. Among P1 cells there are several subgroups, differing in their granule dimensions. P2 apudocytes possess granules of one type with a round core and a narrow rim of cytoplasm. P3 cells are characterized with still larger granules, a very dense core and a narrow rim. In large bronchi some groups are found, consisting of two and more endocrine cells of all three types. In the lungs of the 13-week-old fetus P1 cells are defined and a new type of cells, that contain homogenous granules, characterizing by their small size. In 22 weeks of development in the intrapulmonary bronchi apudocytes with granules specific for Ec-cells are found. NEB consists of cells and islands, possessing polymorphous granules. Various types of apudocytes are defined in large bronchi of the 22-week old fetus. In the stillborn infant apudocytes in the lung are found very seldom. In lung of the mature persons the morphology of apudocytes is unitypical. Thus, during embryogenesis and after birth there are variable types of endocrine cells and NEB. PMID- 2565713 TI - [Duodenal ulcer with gastrin cell hyperplasia--a special form of peptic ulcer]. AB - Pap test was used to study gastric and duodenal G and D cells, blood gastrin levels, basal and stimulated acid production, clinical manifestations, and morphological characteristics of the mucosa in 39 patients with duodenal ulcer and 13 controls. The findings enable the authors to outline a special form of peptic ulcer that is characterized by gastrin cellular hyperplasia concurrent with relative pyloric D cell deficit, gastric metaplasia in the duodenum and gastric hypersecretion. Such patients have more frequently multiple ulcers, predominantly 0(I) blood group, complication-aggravated course of the disease, and ulcer history in close relatives. Moreover, it has been demonstrated that incidence of gastric metaplasia is due to gastric hypersecretion; hyperplasia of duodenal gastrin cells is associated with incidence of gastric metaplasia in patients with peptic ulcer. PMID- 2565714 TI - [Functional morphology of breast APUD cells in dysplasia and neoplastic processes]. AB - Histochemical and immunohistochemical methods were used to examine 29 malignant tumors (18 lobular and 11 invasive carcinomas) and 34 fibroadenomas of the mammary gland (MG). APUD cells containing serotonin, melatonin, and beta endorphine were shown to be present in the duct epithelium of the normal MG and its pericanalicular fibroadenoma. APUD cells were detected in 21 of the 29 malignant tumours of MG. Hormonal differences of APUD cells were found in poorly and well differentiated carcinomas: the former contained serotonin, melatonin, and beta-endorphine (inhibitors of proliferation), the latter--insulin and adrenocorticotropic hormone (stimulators of cell division). Such differences in the endocrine function of MG malignant tumors are likely to be significant in the clinical course and determination of prognosis for carcinomas of various differentiation. PMID- 2565715 TI - Attachment of Moraxella bovis to calf corneal cells and inhibition by antiserum. AB - An in vitro assay was developed using calf corneal cells to assess the importance of fimbriae in the colonisation of the bovine ocular surface by Moraxella bovis, and the role of fimbrial antibodies in the bovine immune response and resistance to infectious bovine keratoconjunctivitis (IBK). Fimbriae promoted adherence of M. bovis to calf corneal cells in culture; 15 fimbriate isolates, representative of 6 fimbrial serogroups of M. bovis, adhered to the cells whereas 4 non fimbriate isolates failed to do so. Fimbrial antibodies in hyperimmune rabbit serum inhibited attachment of all fimbriate strains of the homologous fimbrial serogroup but not those of 5 heterologous serogroups. The relevance of these results to the use of a polyvalent fimbrial vaccine in the control of IBK is discussed. PMID- 2565716 TI - Partitioned twin analysis: a power study. AB - Individual differences in the human genome may now be measured with molecular genetic techniques. Therefore, dizygotic (DZ) twins may be classified as sharing two, one, or zero "genes" identical by descent for any measured polymorphism. As a result, we may partition genetic variation into two sources: (i) genotypes at and closely linked to particular marker loci identified with restriction fragment length polymorphisms (RFLPs) and (ii) other genetic variation. The power of the classical twin study to reject false models lacking either a marker effect or a residual genetic effect is explored. Additivity of genetic effects at or near the locus and of the residual genetic variation as well as random environmental variation are assumed. Results indicate that statistical rejection of models could be achieved with sample sizes which are within the range of several current twin registers. A design including monozygotic (MZ) twins is compared with one consisting of only DZ twins. MZ twins add considerable power for the detection of residual genetic variation but provide no information to resolve genetic marker effects. PMID- 2565717 TI - Evolutionary relationships between laboratory mice and subspecies of Mus musculus based on the restriction fragment length variants of the chymotrypsin gene at the Prt-2 locus. AB - Restriction endonuclease fragment length variants in mice were compared by Southern blot analysis using the cDNA probe pcXP33 for the chymotrypsin gene. The variants were detected in the restriction patterns generated by fragments from digestions with BglII, EcoRI, HindIII, Pstl, SacI, and XbaI. The set of protein phenotypes and the restriction patterns of chymotrypsin gene were examined in many laboratory strains and wild subspecies. Most laboratory strains (26 strains) are grouped into a set defined as Set 1, but only a few laboratory strains (AU/SsJ and five BALB/c sublines) are classified as belonging to Set 2. Of wild subspecies, only BRV-MPL (M. brevirostris) can be placed in Set 1, while DOM-BLG and SK/Cam (M. domesticus) belong in Set 2. The assignment of an appropriate set defined by the characteristics of the chymotrypsin gene has also been investigated in M. musculus, two Chinese subspecies, M. yamashinai, M. molossinus, and M. castaneus, and the evolutionary relationship between laboratory mice and various subspecies of Mus has been examined. PMID- 2565718 TI - Detection of enhancer binding proteins recognizing the human immunodeficiency virus long terminal repeat by in situ gel retardation. AB - We have examined the formation of a DNA "enhancer" - protein complex occurring in situ. Oligonucleotides corresponding to the human immunodeficiency virus (HIV) core enhancer sequence were synthesized, annealed and radiolabeled. The DNA was electroporated either into Jurkat cells or into fresh human peripheral blood T lymphocytes. After the appropriate incubation time and stimulation with various mitogenic agents, cells were lysed and the lysates were electrophoresed on a native polyacrylamide gel. The specific protein-nucleic acid complexes which we obtained were apparently identical to those observed with the "classical" in vitro gel mobility shift assay: one complex seems to be constitutive and the other is induced by mitogens. Additionally competition experiments using "cold" oligonucleotides demonstrated binding specificity in situ. We recommend this novel method for studying DNA-binding proteins and their activation since it requires as few as 10(6) cells, may use primary tissue isolates, and furthermore, allows the rapid assessment of cellular activation signals involved in the post transcriptional modification of trans-acting factors. PMID- 2565719 TI - Expression of surface antigen and mRNA for the CD11c (alpha X, p150) subunit of the human leukocyte adherence receptor family in hematopoietic cells. AB - We characterized the surface antigen and mRNA expression for the CD11c (alpha X, p150) subunit of the human leukocyte adherence receptor family during hematopoietic cell differentiation. The CD11c subunit antigen and mRNA are constitutively expressed in undifferentiated HL-60 promyelocytic leukemia cells, and levels increase markedly with differentiation along the monocyte/macrophage pathway using phorbol myristate acetate. Human monocyte-derived macrophages and human alveolar macrophages express elevated levels of the CD11c subunit antigen and mRNA, indicating that the changes observed in vitro are present in vivo. Dot blot analysis of immature and mature lymphoid and myeloid cells and cell lines demonstrate equivalent levels of CD11c mRNA expression. We conclude that CD11c gene expression is selectively increased during hematopoietic cell differentiation along the monocyte/macrophage pathway. PMID- 2565720 TI - Developmental change and molecular properties of somatostatin receptors in the rat cerebral cortex. AB - The postnatal development and molecular properties of somatostatin receptor were studied in rat cerebral cortex. With [125I-Tyr11]SRIF as a radiolabeled ligand, the specific ligand binding to crude membrane increased transiently in the early phase of postnatal development and then decreased. This increase of somatostatin binding was mainly due to the increased number of binding sites. The two subtypes classified by Tran et al., SSA and SSB, were confirmed and the studies on the relative amount of the subtypes revealed that more SSA subtype was expressed compared with SSB subtype during a week after birth, but, thereafter, both subtypes were almost equally expressed throughout the developmental stages tested. Molecular weight of the covalently labeled somatostatin receptor (SSA subtype), which was determined with the aid of the cross-linking agents, was estimated to be approximately 71,000 with no intramolecular disulfide bond. PMID- 2565722 TI - RFLP studies in different ethnic groups. PMID- 2565721 TI - Lack of correlation between the apolipoprotein B XbaI polymorphism and blood lipid levels in a Swedish population. AB - The possible connections between the apolipoprotein B (apo B) XbaI polymorphism and the serum levels of total cholesterol, triglycerides, LDL, HDL and apo B have been investigated among 187 randomly selected subjects from Gothenburg, Sweden. The interferences of age and sex on the serum lipoproteins and apo B concentrations were considered. Using multiple regression analysis, we compared the different lipid levels and the levels of apo B with the genotypes X1X1, X1X2 and X2X2 (X1 = without the XbaI restriction site, X2 = with the site), with age and with sex and with those factors combined with each other. A significantly higher concentration of serum cholesterol and LDL among men than among women was found and total serum cholesterol, LDL and apo B were positively correlated with age. The allele frequency of the XbaI polymorphism in the sample was 0.45 for the allele without the XbaI restriction site. No correlation was found between the apo B genotypes and the levels of serum lipoproteins or apo B. PMID- 2565723 TI - The knee-chest position does not improve the efficacy of ipecac-induced emesis. AB - Previous studies have shown that ipecac-induced emesis, even if instituted very early, removes only a mean of 28% to 45% of an ingested tracer. Because vomiting is an ancient reflex that occurs in mammals, reptiles, and other animals, we speculated that, in humans, maintaining a sitting rather than a horizontal posture during induced emesis might decrease the efficacy of gastric emptying. To test this hypothesis, 20 normal fasting adult subjects underwent induced emesis in the knee-chest position on one day and in the sitting position on another. Twenty-five 100-micrograms tablets of cyanocobalamin were ingested as a tracer along with 250 mL tap water. Ten minutes after tracer ingestion, 30 mL ipecac syrup and 640 mL tap water were swallowed. All resulting vomitus was homogenized, frozen, and later assayed for cobalt using atomic absorption spectrophotometry. There was no difference in mean tracer recovery with the two positions: knee chest, 47.2% v sitting, 46.9% (paired t test, P greater than .95). Analysis of cobalt recovery for all 40 episodes of emesis revealed a mean of 51.2 +/- 23.7 (SD) micrograms out of 108.7 micrograms total cobalt ingested (95% Cl, 43.6 to 58.7 micrograms). This represented 47.1% of the administered tracer dose (95% Cl, 40.1% to 54.0%). Even if initiated only ten minutes after an ingestion, ipecac induced emesis removes an average of less than half of an ingested tracer dose, with a high degree of intersubject variability. Horizontal patient positioning does not appear to improve the efficacy of this procedure. PMID- 2565724 TI - Rapid tranquilization of the violent patient. AB - Agitated, psychotic patients with the potential for violence pose significant management problems for emergency department staff. With the advent of rapid tranquilization (RT), clinicians were offered a safe, effective method for controlling such patients, eliminating the need for restraints or seclusion rooms. While RT is regarded as a major treatment innovation in psychiatry, nonpsychiatrists are reluctant or unaware of the uses of antipsychotic medication as it pertains to RT. This article provides a brief overview of the pharmacokinetics of antipsychotic medication and reviews the following aspects of RT: route of administration, dosing, time intervals between doses, side effects, and alternative medications for RT. The authors also offer practical guidelines for RT use in the emergency department. PMID- 2565725 TI - Prolonged Q-T interval following astemizole overdose. AB - Astemizole overdose has been reported to cause torsades-de-pointes and in the present case it caused prolongation of the Q-T interval. Astemizole overdoses should be managed in a similar way to overdose with other cardiotoxic drugs. PMID- 2565726 TI - Lymphocyte subpopulation with low membrane potential in the blood of cyclosporin- and prednisone-treated patients: in vivo selectivity for T4 subset. AB - Previous work on the mode of action of CsA revealed that this drug shifts the membrane potential of human periferal blood lymphocytes in vitro. Recently we have analyzed lymphocytes of cyclosporin-treated transplant patients. Membrane potential analysis with the DIOC6(3) fluorescence dye indicates that all the studied patients have a subpopulation of lymphocytes with low membrane potential and that this population is made up predominently of OKT4+ cells. However there is no correlation between the clinical events and the percentage distribution of lymphocyte subpopulations as defined by the membrane potential and the T4/T8 ratio of the total lymphocyte population. PMID- 2565727 TI - [Neurotransmitters and neurologic disorders]. AB - Neurotransmitters should fulfil the following main three criteria; 1) they should be localized in synaptic terminals, 2) they should have certain actions on neurons, 3) they should be released from the terminals after nerve stimulation. In addition to the above three, recent studies have revealed that neurotransmitters should have their own receptors. Moreover, recent immunohistochemical studies have revealed certain neurons to have two or more neurotransmitter candidates; one classic and the other non-classic neurotransmitter candidate. These progresses in the field of neurotransmitter science were briefly reviewed. As an extension of the basic studies mentioned above, clinical studies of measurements of neurotransmitter markers in brains of neurodegenerative disorders are now actively performed. Based on my own studies in the same line, it is possible to summarize the fundamental patterns of neurotransmitter alterations in neurodegenerative disorders as follows: 1) they decrease as a result of degeneration of neuron groups (the most popular pattern); 2) they decrease in spite of the morphologically normal appearance of neurons ("biochemical non-functioning"); 3) they increase in the projecting regions of certain neurons which escaped degeneration. This may be a result of simple condensation of normal nerve terminals or real increase based on the compensatory mechanisms. PMID- 2565728 TI - Effect of Wy-45,727 on 24 h intragastric acidity in twenty normal volunteers. AB - 1. The effects of single daily doses of 20, 50 and 150 mg of WY-45,727, a novel H2-receptor antagonist, and placebo were compared using long term pH-monitoring in 20 male volunteers. 2. Intragastric acidity was measured using combined Ingold glass electrodes. Subjects underwent four studies each under identical dietary conditions. Medication was taken after the evening meal. 3. Median 24 h pH rose from 1.3 (1.2-1.4 interquartile range) on placebo to 1.9 (1.6-2.8) on 20 mg WY 45,727, to 3.1 (2.3-3.7) on 50 mg WY-45,727 and to 4.5 (3.7-4.7) on 150 mg WY 45,727. All three doses increased 24 h and night-time pH significantly compared with placebo (P less than 0.0001). 4. The PAGE test for order effects confirmed clear dose dependent inhibition of acidity (P less than 0.0001). 5. Highly consistent individual responses were found during the night following 150 mg WY 45,727. PMID- 2565729 TI - Importance of beta 2-adrenoceptor stimulation in the suppression of intradermal antigen challenge by adrenaline. AB - 1. Seven atopic subjects received two injections of antigen and one of saline intradermally in the back on each of 4 separate days. They were pretreated with four different drug combinations: (a) adrenaline 0.3 mg subcutaneously over the deltoid muscle (b) subcutaneous adrenaline preceded by 5 mg of the specific beta 2-adrenoceptor antagonist ICI 118,551 orally (c) 8 mg of salbutamol orally (d) placebo. Tablets were given 2 h before and subcutaneous injections 15 min before the intradermal injections of saline and antigen. 2. The median flare response to intradermal low dose antigen and high dose antigen after pretreatment with adrenaline was 4% and 49% of the response seen following pretreatment with placebo (P less than 0.001). When adrenaline was preceded by ICI-118,551, the corresponding median flare responses were 2% and 44% (P less than 0.001) of the placebo response. The flare response after pretreatment with salbutamol was not significantly different from placebo. 3. Adrenaline suppressed the median weal response to the higher dose of antigen to 52% of the response after pretreatment with placebo (P less than 0.05). This suppression by adrenaline was blocked by pretreatment with ICI 118,551. The median weal response after the highest dose of antigen was suppressed by salbutamol to 66% of the response seen after placebo, although this was not significant even when a further three subjects were studied with either salbutamol or placebo.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565730 TI - Long-term study of chimaerism in bone marrow transplantation recipients for severe aplastic anaemia. AB - We used minisatellite probes to analyse by DNA fingerprints the long-term engraftment (median 4.3 years, range 1-2) of 21 bone marrow transplantation recipients for severe aplastic anaemia. Patients received their graft from histocompatible siblings. They were conditioned with cyclophosphamide (150 mg/kg) and a 6GY thoracoabdominal irradiation and did not have ex-vivo T cell depletion of marrow donor. DNA was extracted peripheral mononuclear cells and analysed by Southern blotting with 32P-labelled single-stranded RNA probes. Seven out of 21 donor-recipient pairs were sex-mismatched and additionally studied with a probe detecting a male specific repeated sequence on the Y chromosome. Red cell surface phenotype was also used as marker of engraftment in most cases. Long-term engraftment appeared complete for all patients studied with respect to the three methods. PMID- 2565731 TI - Mobilisation of the tongue by Tunnelierung and split skin graft insertion. AB - Immobility of the tongue, secondary to an illness or subsequent to surgical treatment of orofacial malignancies, reduces the ability of the patient to communicate in a socially acceptable manner. For some specific anatomical situations, a new technique is described allowing uncomplicated tongue mobilisation and restoration of its function. PMID- 2565732 TI - Mercaptan and dicarboxylate inhibitors of hamster dihydroorotase. AB - In mammals, dihydroorotase is part of a trifunctional protein, dihydroorotate synthetase, which catalyzes the first three reactions of de novo pyrimidine biosynthesis. Dihydroorotase catalyzes the formation of a peptide-like bond between the terminal ureido nitrogen and the beta-carboxyl group of N-carbamyl-L aspartate to yield heterocyclic L-dihydroorotate. A variety of evidence suggests that dihydroorotase may have a catalytic mechanism similar to that of a zinc protease [Christopherson, R. I., & Jones, M. E. (1980) J. Biol. Chem. 255, 3358 3370]. Tight-binding inhibitors of the zinc proteases, carboxypeptidase A, thermolysin, and angiotensin-converting enzyme have been synthesized that combine structural features of the substrates with a thiol or carboxyl group in an appropriate position to coordinate a zinc atom bound at the catalytic site. We have synthesized (4R)-2-oxo-6-thioxohexahydropyrimidine-4-carboxylate (L-6 thiodihydroorotate) and have found that this analogue is a potent competitive inhibitor of dihydroorotase with a dissociation constant (Ki) in the presence of excess Zn2+ ion of 0.17 +/- 0.02 microM at pH 7.4. The potency of inhibition by L 6-thiodihydroorotate in the presence of divalent metal ions decreases in the order Zn2+ greater than Ca2+ greater than Co2+ greater than Mn2+ greater than Ni2+; L-6-thiodihydroorotate alone is less inhibitory and has a Ki of 0.85 +/- 0.14 microM. 6-Thioorotate has a Ki of 82 +/- 8 microM which decreases to 3.8 +/- 1.4 microM in the presence of Zn2+. Zn2+ alone is a moderate inhibitor of dihydroorotase and does not enhance the potency of other inhibitors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565733 TI - Changes in the cofactor binding domain of bovine striatal tyrosine hydroxylase at physiological pH upon cAMP-dependent phosphorylation mapped with tetrahydrobiopterin analogues. AB - The structure of the cofactor binding domain of tyrosine hydroxylase (TH) was examined at physiological pH by determining kinetic parameters of (R) tetrahydrobiopterin [(R)-BH4] and a series of tetrahydropterin (PH4) derivatives (6-R1-6-R2-PH4: R1 = H and R2 = methyl, hydroxymethyl, ethyl, methoxymethyl, phenyl, and cyclohexyl; R1 = methyl and R2 = methyl, ethyl, propyl, phenyl, and benzyl). A minimally purified TH preparation that was not specifically phosphorylated (designated as "unphosphorylated") was compared with enzyme phosphorylated with cAMP-dependent protein kinase. The Km for tyrosine with most tetrahydropterin analogues ranged between 20 and 60 microM with little decrease upon phosphorylation. Two exceptions were an unusually low Km of 7 microM with 6 ethyl-PH4 and a high Km of 120 microM with 6-phenyl-6-methyl-PH4, both with phosphorylated TH. Tyrosine substrate inhibition was elicited only with (R)-BH4 and 6-hydroxymethyl-PH4. With unphosphorylated TH (with the exception of 6-benzyl 6-methyl-PH4, Km = 4 mM) an inverse correlation between cofactor Km and side chain hydrophobicity was observed ranging from a high with (R)-BH4 (5 mM) to a low with 6-cyclohexyl-PH4 (0.3 mM). An 8-fold span of Vmax was seen overall. Phosphorylation caused a 0.6-4-fold increase in Vmax and a 35-2000-fold decrease in Km for cofactor, ranging from a high of 60 microM with 6-methyl-PH4 to a low of 0.6 microM with 6-cyclohexyl-PH4. A correlation of the size of the hydrocarbon component of the side chain with affinity is strongly evident with phosphorylated TH, but in contrast to unphosphorylated enzyme, the hydroxyl groups in hydroxymethyl-PH4 (20 microM) and (R)-BH4 (3 microM) decrease Km in comparison to that of 6-methyl-PH4. Although 6,6-disubstituted analogues were found with affinities near that of (R)-BH4 (e.g., 6-propyl-6-methyl-PH4, 4 microM), they were frequently more loosely associated with phosphorylated TH than their monosubstituted counterparts (6-phenyl-PH4, 0.8 microM; cf. 6-phenyl-6-methyl PH4, 8 microM). A model of the cofactor side-chain binding domain is proposed in which a limited region of nonpolar protein residue(s) capable of van der Waals contact with the hydrocarbon backbone of the (R)-BH4 dihydroxypropyl group is opposite to a recognition site for hydroxyl(s). Although interaction with either the hydrophilic or hydrophobic regions of unphosphorylated tyrosine hydroxylase is possible, phosphorylation by cAMP-dependent protein kinase appears to optimize the simultaneous operation of both forces. PMID- 2565734 TI - Complexation of fibronectin with tissue transglutaminase. AB - Previous work [Lorand, L., Dailey, J. E., & Turner, P. M. (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 1057-1059] showed that fibronectin might serve as a specific carrier for transglutaminases accidentally discharged from erythrocytes or other cells into plasma. In the present study we examined the association of these proteins in purified systems. Complexation was readily demonstrable by nondenaturing electrophoresis, using dansylcadaverine-dependent activity staining as well as immunoblotting procedures, and also by HPLC gel filtration. The results indicate a stoichiometry of 2:1 for the binding of the human erythrocyte transglutaminase (80K) to human plasma fibronectin (440K). The attachment is noncovalent in nature and does not involve cross-linking of the proteins either to themselves or to each other. Binding occurs in the absence of Ca2+, suggesting that a domain on the transglutaminase molecule other than the catalytic site is needed for complexation with fibronectin. Limited proteolysis with chymotrypsin for delineating the relevant region in fibronectin yielded two gelatin- (collagen) binding fragments (56K and 46K), each displaying affinity for transglutaminase. Moreover, these fragments--like intact fibronectin--bound erythrocyte transglutaminase and gelatin simultaneously in ternary complexes. PMID- 2565735 TI - Activities of enzymes of lipid metabolism in Morris hepatoma 7800 C1 cells. AB - (1) The rate of palmitate oxidation in the 7800 C1 Morris hepatoma cells was about 60% of the activity observed in hepatocytes. The stimulatory effect of glucagon in hepatocytes was not observed in the hepatoma cells. The rate of fatty acid synthesis from [2-14C]acetate in the hepatoma cells was 1/20 of the activity in hepatocytes. The conversion of [2-14C]acetate to cholesterol was not different in the two kinds of cell. (2) Acetyl-CoA carboxylase and fatty acid synthetase were significantly decreased in the hepatoma cells. The hepatoma cells had, however, raised activities of malate dehydrogenase (decarboxylating), and glucose 6-phosphate and 6-phosphogluconate dehydrogenases. (3) The activities of the enzymes were not affected by different concentrations of glucose or palmitate in the culture medium. Insulin, dexamethasone, triiothyronine and glucagon had no effect on the enzyme activities. This is in contrast to the adaptation of the peroxisomal beta-oxidation system, which is induced by fatty acids and modified by hormones. PMID- 2565736 TI - The effect of methionine on methotrexate metabolism in rat hepatocytes in monolayer culture. AB - The effect of methyl donors on the metabolism of methotrexate has been investigated in rat hepatocytes in monolayer culture. Pulse exposure to low concentrations of methotrexate (1 microM, 3h) in the absence of methionine results in the facile formation of the di- to pentaglutamates with the di- and triglutamate predominating. Further incubation after the removal of methotrexate (MTX) results in a shift to the tetra- and pentaglutamate at the expense of the shorter chain length derivatives. The same measurement in the presence of 1 mM methionine causes approx. an 80% inhibition in the formation of polyglutamates. This effect can be partially achieved when methionine is replaced by choline or betaine. No alteration in the formation of 7-hydroxymethotrexate could be detected by similar changes in methionine concentrations in the medium. The activity of the enzymes which synthesize and degrade methotrexate polyglutamates, folylpolyglutamate synthetase and gamma-glutamyl hydrolase, respectively, were the same in extracts of cells grown in the absence and in the presence of 1 mM methionine. Incubation of the hepatocytes with methionine causes a significant increase in 5,6,7,8-tetrahydrofolate (H4folate), 5,10-methylenehydrofolate and 10 formyltetrahydrofolate and a decrease in 5-methyltetrahydrofolate. These results suggest that the inhibition of glutamylation of methotrexate could be due in part to an elevation in reduced folates which can more effectively compete with methotrexate as a substrate for folylpolyglutamate synthetase. Inhibition in methotrexate glutamylation by methionine, betaine and choline in hepatocytes may contribute to the alleviation of hepatic toxicity by methyl donors. PMID- 2565737 TI - Purification and properties of DAHP synthase from Nocardia mediterranei. AB - 3-Deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase, the first enzyme of the shikimate pathway was isolated from Nocardia mediterranei. It has a molecular weight of approx. 135,000, and four identical subunits, each with a molecular weight of 35,000. The Km values for phosphoenolpyruvate (PEP) and D-erythrose 4 phosphate (E-4-P) were 0.4 and 0.25 mM, respectively, and kinetic study showed that LTrp inhibited DAHP synthase activity, but was not competitive with respect to PEP or E-4-P. The enzyme activity was inhibited by excess of E-4-P added in the incubation system. D-ribose 5-phosphate (R-5-P), D-glucose 6-phosphate (G-6 P) or D-sedoheptulose 7-phosphate (Su-7-P) etc. inhibited DAHP synthase in cell free extract, but on partially purified enzyme no inhibitory effect was detected. The indirect inhibition of R-5-P and other sugar phosphates was considered to be due to the formation of E-4-P catalyzed by the related enzymes present in cell free extract. PMID- 2565738 TI - [Human thrombocyte guanylate cyclase in diabetes mellitus]. AB - Clear differences in properties of platelet guanylate cyclase from healthy donors and patients with diabetes mellitus were identified; departure from the norm was more pronounced in the case of the II-type of this disease, than the I-type. We have registered the decrease in the Mg-activity (basal) of guanylate cyclase by 30 and 50 per cent, Mn-activity--by 20 and 30 per cent, the state of guanylate cyclase activation by 0.1 mM sodium nitroprusside to 2 and 3-fold in patients with I and II-type of diabetes mellitus, consequently. The possible causative factors of these disturbances are discussed. It is suggested that the decrease in guanylate cyclase activation by nitroprusside is due to the enzyme heme deficiency rising with the diabetes mellitus. PMID- 2565739 TI - Correlation between interleukin-1 production and engraftment of transplanted bone marrow stem cells in patients with lethal immunodeficiencies. AB - Interleukin-1 (IL-1) production by endotoxin-stimulated, cultured monocytes from 19 patients with lethal congenital immune disorders were studied and compared with normal controls. Lipopolysaccharide (LPS) stimulated IL-1 production was normal in three of three patients with Wiskott Aldrich syndrome (WAS), two of three combined immunodeficiency with T-cell predominance (CIDTP) and nine of 13 with severe combined immunodeficiency (SCID). Monocytes deficient in IL-1 production could be restored to normal production after incubation with indomethacin in three of five deficient patients. Monocytes from the other two patients could not be induced to generate IL-1, suggesting either an intrinsic deficiency or an alternate inhibitory mechanism as the basis for the IL-1 deficiency observed. In patients with SCID who were transplanted with HLA haplotype disparate, T-cell depleted marrow without preparative chemotherapy, deficiency of monocyte IL-1 production was correlated with graft failure. Immune reconstitution was achieved in IL-1 deficient patients only when donor monocytes were also engrafted. We hypothesize that deficiencies of IL-1 production may contribute to the heterogeneous expression of combined immunodeficiencies, and may also restrict the engraftment and functional development of allogeneic lymphoid progenitors from a T-cell depleted marrow graft. PMID- 2565740 TI - The murine M-CSF gene is localized on chromosome 3. PMID- 2565741 TI - Origin of leukemic relapse after bone marrow transplantation: comparison of cytogenetic and molecular analyses. AB - Leukemic relapse following bone marrow transplant (BMT) is generally due to the recurrence in recipient cells, but may rarely occur as a result of donor cell transformation. Donor cell relapse is generally identified using cytogenetic markers such as the sex chromosomes. Recently, molecular techniques have been used to identify the origin of bone marrow cells by their DNA restriction fragment length polymorphisms. We describe the case of a male pediatric patient who had a leukemic relapse 30 months following BMT from his sister. Both cytogenetic and molecular techniques were used to identify the origin of the leukemic relapse. Cytogenetic analyses indicated the absence of the Y chromosome and the presence of a donor cell type 9qh polymorphism, suggesting a donor cell relapse. Molecular analyses also indicated the absence of the Y chromosome but demonstrated the recurrence of recipient DNA markers from three other chromosomes, suggesting a recipient cell relapse. While the leukemic cell lineage cannot be definitively assigned in this case, our results suggest that caution must be exercised when assigning leukemic cell lineage following post-BMT relapse. PMID- 2565742 TI - Characterization of seven antihistamines, their N-oxides and related metabolites by fast atom bombardment mass spectrometry and fast atom bombardment tandem mass spectrometry. AB - We have examined the synthetic N-oxides of five ethylenediamine-type antihistamines using fast atom bombardment (FAB) mass spectrometry and FAB tandem mass spectrometry (MS/MS). Fragmentation of the protonated molecule in the normal and collisionally activated spectra appeared to be characteristic for this class of antihistamine N-oxide. Spectra were also acquired from an ethanolamine and a propylamine antihistamine N-oxide for comparison. These results were very similar to those obtained from biologically produced antihistamine N-oxides, as well as isomeric metabolites, which were readily distinguished from the N-oxides by characteristic fragmentation. In addition, a prominent ion 16 daltons lower in mass, which has been attributed to loss of elemental oxygen from the protonated N oxide in chemical ionization mass spectral studies, was shown to be a matrix dependent product of the solution-phase reduction of the antihistamine N-oxide to the parent antihistamine during FAB ionization. These results demonstrate that with a non-reducing matrix such as glycerol, FAB mass spectrometry and FAB MS/MS are excellent methods for the characterization of the non-conjugated antihistamine metabolites such as the N-oxides. PMID- 2565743 TI - Profound bradycardia after addition of diltiazem to a beta blocker. PMID- 2565744 TI - Treatment of chronic granulocytic leukemia in blastic crisis. PMID- 2565745 TI - Genetics and biology of human retroviruses. PMID- 2565746 TI - The role of peripheral blood stem cells as rescue after myeloablative therapy. PMID- 2565747 TI - From radiation chimeras to 1988. PMID- 2565748 TI - Uses and abuses of benzodiazepines. PMID- 2565749 TI - Blockade of both D1- and D2-dopamine receptors inhibits amphetamine-induced ascorbate release in the neostriatum. AB - In vivo recordings with electrochemically modified microvoltammetric electrodes revealed that several neuroleptic drugs, including haloperidol, clozapine, and thioridazine, blocked the rise in extracellular ascorbate produced by amphetamine in the neostriatum of urethane-anesthetized rats. This effect was also observed in animals that received a combined injection of Sch-23390 and sulpiride, but not when either of these drugs were administered alone or in combination with the 5 HT2 blocker, ritanserin. These results indicate that a combined blockade of D1- and D2-dopamine receptors blocks amphetamine-induced ascorbate release. PMID- 2565750 TI - A conformationally restricted analogue of L-glutamate, the (2S,3R,4S) isomer of L alpha-(carboxycyclopropyl)glycine, activates the NMDA-type receptor more markedly than NMDA in the isolated rat spinal cord. AB - Depolarizing actions of 4 conformationally restricted L-glutamate analogues, (2S,3S,4S) isomer (L-CCG-I), (2S,3R,4R) isomer (L-CCG-II), (2S,3S,4R) isomer (L CCG-III) and (2S,3R,4S) isomer (L-CCG-IV) of L-alpha-(carboxycyclopropyl)-glycine (L-CCG), were investigated in the isolated rat spinal cord by extracellular recordings of potential changes of motoneurones from the ventral roots, in order to study the interaction between the conformation of glutamate and its receptor subtype. The order of the depolarizing activity was quisqualate greater than L CCG-IV = kainate greater than NMDA greater than L-CCG-I greater than L-CCG-III greater than L-CCG-II. The depolarization caused by L-CCG-IV was effectively blocked by the NMDA antagonists and Mg2+ ions, while the L-CCG-I response was not affected by these blockers. These results suggest that the NMDA-type receptor is activated by a folded form of L-glutamate. PMID- 2565751 TI - Actions of somatostatin on perfused bovine adrenal glands and cultured bovine adrenal medullary cells. AB - The effects of somatostatin on catecholamine secretion and inositol phosphate accumulation have been studied using isolated perfused bovine adrenal glands and cultured bovine adrenal medullary cells. Somatostatin had no effect on basal adrenaline or noradrenaline secretion from either preparation. At concentrations above 1 microM, somatostatin inhibited the secretion of both catecholamines induced by 5 microM nicotine from cultured chromaffin cells. In contrast, over the concentration range 0.1 nM-10 microM, somatostatin had no effect on the secretory responses produced by 10 nM angiotensin II or 1 microM histamine. Inositol phosphate accumulation in cultured bovine adrenal medullary cells was unaffected by 0.1 nM-0.1 microM somatostatin, however at 1 and 10 microM somatostatin it was significantly increased, by 23% and 103% respectively. The effects of somatostatin (0.1 nM-10 microM) and of 50 microM muscarine on inositol phosphate accumulation were simply additive. Similarly, somatostatin at 0.1 nM and 10 nM together with 10 nM angiotensin II or 1 microM histamine produced additive inositol phosphate responses. In contrast, 1 microM somatostatin gave significantly more-than-additive (synergistic) inositol phosphate responses with angiotensin II and histamine. The results suggest that some adrenal medullary cells possess several types of receptors, and that these receptors may interact to produce non-additive responses. PMID- 2565752 TI - GM1 ganglioside-induced recovery of nigrostriatal dopaminergic neurons after MPTP: an immunohistochemical study. AB - The administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to mice results in the loss of dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) from the mouse striatum and a loss of cells containing tyrosine hydroxylase (TH) immunoreactivity from the substantia nigra. The cells that remained in the nigra after MPTP treatment were smaller in diameter than normal cells. Treatment with GM1 ganglioside beginning 24 h after establishing the MPTP lesion resulted in partial restoration of DA and DOPAC content in the striatum and an increase in the diameter of the TH-immunoreactive nigra cells. It appears, therefore, that treatment of MPTP-intoxicated mice with GM1 ganglioside results in the partial restoration of both the biochemistry and morphology of dopaminergic neurons. PMID- 2565753 TI - Effects of intrahypothalamic injections of GABA, muscimol, pentobarbital, and L glutamic acid on feed intake of satiated sheep. AB - Five wethers were surgically prepared with cranial implants to study the role of gabaminergic neural pathways on the hypothalamic control of feeding behaviour in ruminants. In the first experiment, the animals were injected (1 microL) with a physiological Tyrode (0.95%) solution, muscimol (0.5 and 1.0 nmol), GABA (0.5 and 1.0 nmol), and L-glutamic acid (0.5 and 1.0 nmol). Feed intake following injections of muscimol (1.0 nmol) and L-glutamic acid (0.5 and 1.0 nmol) was twice as large as that following the Tyrode solution, at 60-min postinjections. These results, however, were not statistically significant (p = 0.12-0.15). In the second experiment, the animals were injected (1 microL) with saline, muscimol (0.8 nmol), L-glutamic acid (0.8 nmol), and pentobarbital (0.26 mumol). Fifteen minutes after the injections, pentobarbital had induced a significant feeding response when compared with control values (p less than 0.01), whereas the effect of L-glutamic acid was not significant. However, 30 min after the injections, feed intake of sheep having received L-glutamic acid was higher than that obtained with the control injections (p less than 0.01). The response to pentobarbital was stronger than that to either muscimol or L-glutamic acid. Histological analyses of brain tissue indicated that injections were performed in the ventromedial hypothalamus of four sheep and in the dorsomedial hypothalamus of the other. The data indicate that L-glutamic acid stimulates feed intake by acting either as a precursor of GABA or by a direct stimulation of glutaminergic neural pathways involved in the control of feed intake. PMID- 2565754 TI - Responses of the isolated rectum of the rainbow lizard (Agama agama) to sympathomimetics. AB - The relaxant effects of isoprenaline, noradrenaline, and adrenaline on the isolated rectum of the rainbow lizard (Agama agama) were studied. Responses were measured as a reduction of carbachol-induced contractions for each sympathomimetic agent. Isoprenaline, adrenaline, noradrenaline produced a dose dependent relaxation of this preparation and the order of potency was as given. The pD2 value of 8.15 +/- 1.88 obtained for isoprenaline was significantly different (p less than 0.05) from those for adrenaline (5.80 +/- 0.90) and noradrenaline (5.25 +/- 1.18). H35/25, propranolol, and practolol competitively antagonized the relaxant effects of isoprenaline on the isolated lizard rectum. The pA2 values for these beta-adrenoceptor antagonists did not differ significantly (at p less than 0.05). alpha-Adrenoceptor antagonists, phentolamine and phenoxybenzamine, failed to alter the relaxant responses of these sympathometics to any appreciable extent. These results are interpreted to suggest that the relaxant effect produced by these sympathomimetics are mediated predominantly by beta-adrenoceptors that are not significantly differentiated into subtypes. alpha-Adrenoceptors in this preparation contribute minimally to the observed inhibitory response following sympathomimetic stimulation. PMID- 2565755 TI - Propranolol antagonizes hypotension induced by alpha-blockers but not by sodium nitroprusside or methacholine. AB - A pressor response has been observed with propranolol, a nonselective beta adrenoceptor antagonist, in animals given a nonselective alpha-adrenoceptor antagonist. This study investigates whether a pressor response to propranolol occurs in conscious unrestrained rats following a hypotensive response induced by phentolamine (nonselective alpha-antagonist), prazosin (selective alpha 1 antagonist) and (or) rauwolscine (selective alpha 2-antagonist), sodium nitroprusside (smooth muscle relaxant), or methacholine (muscarinic agonist). The rats were subjected to a continuous infusion of a hypotensive agent or normal saline followed by i.v. injection of propranolol. The infusion of phentolamine significantly decreased mean arterial pressure (MAP). Subsequent injection of propranolol restored MAP to the control level. Prazosin and rauwolscine each caused a small but not significant decrease in MAP which was reversed by propranolol. Concurrent infusions of prazosin and rauwolscine caused a significant decrease in MAP. Subsequent injection of propranolol caused a large pressor response which increased MAP to 20% above control MAP prior to the administration of drugs. Nitroprusside or methacholine each caused a significant decrease in MAP, but the hypotension was not antagonized by propranolol. The concurrent infusions of a low dose of nitroprusside and prazosin caused a significant decrease in MAP which was reversed by propranolol. The infusion of saline did not alter MAP, and propranolol did not cause a pressor response. It is concluded that propranolol antagonizes the hypotensive effect of an alpha-blocker but not that of sodium nitroprusside or methacholine. Our results suggest the presence of a specific interaction between alpha- and beta-antagonists. PMID- 2565756 TI - Presence of bacterial glycocalyx and fimbriae on Pasteurella haemolytica in feedlot cattle with pneumonic pasteurellosis. AB - This investigation was conducted to determine if Pasteurella haemolytica within feedlot cattle affected by pneumonic pasteurellosis express fimbriae (pili) and bacterial glycocalyx. Bacteriological culture of pulmonary tissue from three calves with fibrinous pneumonia resulted in heavy growth of P. haemolytica. Transmission electron microscopy of the lungs showed numerous microcolonies of gram-negative bacteria with morphology typical of Pasteurella haemolytica. The cells within these microcolonies possessed bacterial glycocalyces which stained with ruthenium red. Glycocalyx-encased microcolonies were also present in specimens examined by scanning electron microscopy. Typical P. haemolytica cells were evident in a tracheal specimen and these bacteria had radial glycocalyces consistent with polysaccharide and proteinaceous material condensed on linear structures suggestive of fimbriae. The pathogenetic importance of the bacterial glycocalyx and fimbriae in shipping fever pneumonia has yet to be established but their presence in clinical cases of Pasteurella pneumonia in feedlot cattle further supports a possible role in the initiation and progression of this disease as well as bacterial resistance to antimicrobial agents. PMID- 2565757 TI - Effect of lenperone hydrochloride on gastroesophageal sphincter pressure in healthy dogs. AB - Treatment of healthy dogs with the butyrophenone derivative, lenperone hydrochloride, at two different doses significantly decreased gastroesophageal sphincter pressure (GESP). No dose-related effect was identified. Individual variation in the response to lenperone hydrochloride was noted which was consistent on a day-to-day basis. Lenperone hydrochloride is unsuitable for chemical restraint of dogs undergoing esophageal manometry because it decreases GESP and because the magnitude of the decrease varies considerably between dogs. PMID- 2565758 TI - Recommendations from the Canadian Hypertension Society Consensus Conference on the Pharmacologic Treatment of Hypertension. PMID- 2565761 TI - Alteration of the c-mos locus in "normal" tissues from humans exposed to radium. AB - The structure of a number of human protooncogenes of persons with internal systemic exposure to radium was analyzed by restriction enzyme digestion and Southern blotting of their DNA. Two extra c-mos EcoRI restriction fragment length bands of 5.0 and 5.5 kilobases were found in tissue DNA from six of seven such individuals. The extra c-mos bands were detected in DNA from many, but not all, of the tissues of the individuals exposed to radium. Kidney DNA, however, from three of four individuals exposed to radium contained these alterations; kidney DNA from six age-matched controls did not. The 5.0- and 5.5-kilobase bands, which were of a similar intensity, varied in their intensity with respect to that of the normal 2.5-kilobase band of the c-mos gene. The DNAs that have the polymorphic bands also appear to have a more complex c-mos methylation pattern. Our results suggest that the c-mos restriction fragment length alterations found in individuals exposed to radium were induced rather than inherited, are epigenetic in origin, and most likely result from changes in the methylation of bases surrounding the single exon of the c-mos protooncogene. PMID- 2565759 TI - Anorexia nervosa and bulimia nervosa. AB - No definitive therapy exists for anorexia nervosa (AN) or bulimia nervosa (BN). Nevertheless, biologic and psychologic research into these disorders has increased over the last decade. We examine the various drugs available for treatment. Advances in pharmacotherapy for AN have been modest and have reflected efforts either to stimulate hunger and weight gain or to control complications of the starvation process. Food remains the "drug" of choice. Antidepressants have been found to be beneficial in the treatment of BN. The meaning of this in the context of a relation between BN and mood disorders remains unclear, since coexistent depression does not predict a positive response to these drugs. Pharmacotherapy represents a single but important dimension of the management of patients with eating disorders. The optimal integration of drug therapy and psychotherapy and the identification of predictors of a positive response to drugs have yet to be addressed by clinical research. PMID- 2565760 TI - Induction of rat liver gamma-glutamyltranspeptidase-positive foci by oral administration of 1-nitropyrene. AB - In the present study, the question of whether the ubiquitous environmental pollutant, 1-nitropyrene (1-NP), can induce gamma-glutamyltranspeptidase (GGT) positive foci, an early lesion occurring during hepatocarcinogenesis, when given orally to F344 rats was examined. Significant induction of GGT-positive foci was observed with all the doses of 1-NP (1000, 500, 250 and 100 mg/kg body wt) used in the present experiment after 6 repeated intragastric (i.g.) intubations when accompanied by partial hepatectomy (PH) performed midway, but not after a single i.g. 1000 mg/kg body wt intubation plus PH. The potential for induction of foci by 1-NP, however, was far less than that by benzo[a]pyrene (B[a]P). The results thus suggested a weak but substantial initiation activity for 1-NP in the rat liver when given orally, particularly with repeated doses. PMID- 2565762 TI - Loss of the same alleles of HRAS1 and D11S151 in two independent pancreatic cancers from a patient with multiple endocrine neoplasia type 1. AB - The DNAs from two independent pancreatic cancers (tumors 1 and 2) in a patient with multiple endocrine neoplasia type 1 were analyzed. No amplification or gross rearrangement of 19 protooncogenes was observed. However, Southern blot analysis using polymorphic DNA probes revealed loss of heterozygosity at loci on chromosome 11p in both tumors. In tumor 1, an extensive region including the HRAS1, PTH, CALCA, and D11S151 loci was deleted, while in tumor 2 loss of heterozygosity was limited at the HRAS1 and D11S151 loci. Because loss of heterozygosity at other chromosomal loci in the two tumors was quite rare, loss of genes on 11p might be nonrandom. It is noteworthy that the same allele at the HRAS1 locus and also the same allele at the D11S151 locus were lost in the two independent tumors. These results suggest that loss of genes at the HRAS1 and/or D11S151 loci plays an important role unmasking the remaining sequences probably having a recessive mutation. PMID- 2565763 TI - Characterization of the multidrug resistance protein expressed in cell clones stably transfected with the mouse mdr1 cDNA. AB - Structural features of the multidrug resistance protein encoded by the mouse mdr1 gene were studied in multidrug-resistant cell clones stably transfected with a biologically active cDNA clone. Independently derived transfectant cell clones, initially selected in Adriamycin, were shown to be cross-resistant to several drugs, including actinomycin D, amsacrine, mitoxantrone, VP-16, and vinblastine but remained sensitive to cis-platinum, 5-fluorouracil, arabinocytosine, and bleomycin. In drug-resistant transfectants the mdr1 gene product was greatly overexpressed as a polypeptide of apparent molecular weight 160,000-170,000. This protein was present in membrane enriched fractions and could be metabolically labeled with [3H )glucosamine, confirming that the transfected mdr1 gene encodes a membrane glycoprotein. The protein was found phosphorylated on serine residues and was shown to be photolabeled by both the calcium antagonist azidopine and the ATP analogue 8-azido ATP. Tryptic mapping of the ATP-photoaffinity labeled protein indicated that ATP crosslinking was site-specific and limited to two discrete peptide fragments of the protein, suggesting that the overexpressed mdr protein is capable of direct and specific ATP binding. PMID- 2565764 TI - P-glycoprotein expression in multidrug-resistant human ovarian carcinoma cell lines. AB - Multiple selections with either vinblastine or vincristine in the human ovarian carcinoma cell line SKOV3 resulted in variants with increasing degrees of multidrug resistance. SKOV3 derivatives that span a wide range in resistance (4- to 2000-fold) were obtained and analyzed for P-glycoprotein expression. In general, we observed a progressive increase in P-glycoprotein level (detected by Western blot) that paralleled the increase in multidrug resistance. However, a more detailed analysis of the P-glycoprotein mRNA and gene level indicated that the amount of P-glycoprotein expressed may be under complex control. At low levels of resistance, only an increase in P-glycoprotein mRNA and protein was observed. At intermediate to high levels of resistance P-glycoprotein gene amplification became evident. At the high level of resistance, an example was observed where only the amount of P-glycoprotein was increased without a concomitant increase in mRNA or gene copy. The mechanisms through which the content of P-glycoprotein in the plasma membrane is mediated are not understood; it is possible that the resistant variants identified here represent perturbations at different levels of regulation. PMID- 2565765 TI - A strategy for measuring the therapeutic properties of new antiparkinsonian drugs, such as +PHNO, in patients with on-off fluctuations. AB - We describe a simple quantitative bedside method for assessing the dopaminergic properties of new therapeutic agents, namely, by documenting their duration of action in patients with Parkinson's disease who exhibit "on-off" fluctuations. This model represents a form of human dopaminergic "bioassay". PMID- 2565766 TI - The antiparkinsonian activity of CQA 206-291, a new D2 dopamine receptor agonist. AB - CQA 206-291, a new D2 dopamine receptor agonist with a biphasic dopaminergic profile, was given to six patients with idiopathic Parkinson's disease after overnight drug withdrawal. With incremental single oral doses of CQA, a dose related, clinically significant, and prolonged antiparkinsonian effect was observed. Most subjects experienced drowsiness after the drug while a minority of subjects experienced nausea and/or vomiting or postural hypotension. Further study of this drug in humans is indicated. PMID- 2565767 TI - Alpidem, a novel anxiolytic drug. A double-blind, placebo-controlled study in anxious outpatients. AB - The anxiolytic activity of alpidem (150 mg/day) and its effects on psychomotor performances were compared with placebo in 60 outpatients. The trial was a double blind, parallel group, and the two treatments were administered orally in three divided doses for 3 weeks. Eighteen male and 42 female patients (mean age, 39.6 years) suffering from generalized anxiety or adjustment disorder with anxious mood of at least 1-month duration entered the trial at the end of a 1-week placebo run-in period designed to exclude early placebo responders. Efficacy was assessed with the Hamilton rating scale for anxiety (HRSA), the state-trait anxiety inventory (STAI x 1: anxiety as state), a visual analogue scale (VAS), and clinical global impression (CGI). Psychomotor performance was assessed by the digit symbol substitution test (DSST). Alpidem was significantly more effective than placebo in decreasing the severity of anxiety, both in the physician's judgment [total HRSA (p = 0.007), psychic symptoms (p = 0.0040), somatic symptoms (p = 0.0002)] and in the patients' evaluation [STAI x 1 (p = 0.0001) and VAS (p = 0.0003)]. Psychomotor performance was improved by both treatments; there was no difference between results with alpidem and placebo at the DSST (p = 0.2801), but the improvement was almost twofold on alpidem. Side effects were negligible with both treatments and the efficacy index, obtained from the CGI, was significantly better with alpidem than with placebo after day 7 (at least p less than 0.03). PMID- 2565768 TI - Beta-adrenergic blockers in patients with acute myocardial infarction. AB - Beta-blockers given within the first 24 hours of an acute MI, first as an intravenous bolus followed by oral therapy, have been shown to reduce in-hospital mortality following a myocardial infarction. For the long-term, beta-blockers represent the only documented effective prophylactic treatment for MI patients. The reduction in all-cause mortality is due primarily to a reduction in atherosclerotic cardiovascular deaths, particularly sudden cardiac deaths. This finding is consistent with the experimental observation that beta-blockers raise the threshold for ventricular fibrillation. Indeed, MI patients with complex ventricular arrhythmias respond very favorably to beta-blockers. However, reductions in nonsudden deaths and nonfatal reinfarctions have also been observed, suggesting that the beneficial effects of beta-blockers are not limited to antiarrhythmic effects alone and that these drugs may also have anti-ischemic effects. The prime candidates for beta-blocker therapy are the high-risk MI patients with transient electrical and/or pump complications during the acute phase. If therapy is initiated within hours of an acute infarction, it seems reasonable to continue it after hospital discharge. Evidence from the large, long term trials suggest that, in the absence of any troublesome adverse reaction and given that most post-MI patients experience ventricular arrhythmias and angina, it seems reasonable not to limit the duration of treatment. PMID- 2565770 TI - Proliferating cell nuclear antigen (PCNA/cyclin) immunocytochemistry as a labeling index in mouse lung tissues. AB - Proliferating cell nuclear antigen is expressed in cells from late G1 through the S-phase of the cell cycle. Therefore, antibodies directed against this molecule should provide a probe for labeling immunocytochemically the nuclei of proliferating cells. Herein we demonstrate the feasibility and reliability of this technique by quantifying immunostained pulmonary nuclei. We applied polyclonal and monoclonal antisera to alveolar and bronchiolar pulmonary epithelial cells in various proliferative states in tissue-sections and in vitro. A/J mice had a slightly higher labeling index than C57BL/6J mice, and proliferation in both strains increased dramatically after butylated hydroxytoluene treatment produced compensatory hyperplasia of Type-II pneumocytes. Immunostaining in fetal and neonatal lung samples from mice was higher than in adults. Spontaneous lung adenomas had a higher labeling index than the surrounding normal lung tissue. In addition, new data contained herein demonstrate a strain difference in proliferation of bronchiolar epithelial cells, and quantify the extent to which BHT-induced lung damage increases these proliferative rates. This mammalian nuclear antigen did not cross-react with antiserum to a functionally related bacterial protein, the beta subunit of E. coli DNA polymerase-III holoenzyme. PMID- 2565771 TI - Characterization of gamma-glutamyl transpeptidase activity of cultured endothelial cells from porcine brain capillaries. AB - Endothelial cells were isolated with high viability (greater than 93%) from porcine brain capillaries by Percoll gradient centrifugation after purely enzymatic digestion. Primary cultures were grown to confluent cell monolayers and quantitated for the activity of gamma-glutamyl transpeptidase. The gamma-glutamyl transpeptidase activity starts from a high enzymatic level, decreases with time in culture to about 15% of the initial value, and remains constant at this level after day 10 in culture. The activity progression depends on surface conditions. In the presence of collagen, an exponential decrease starts immediately after seeding, with a time constant of 70 +/- 10 h. In the absence of collagen, gamma glutamyl transpeptidase activity first decreases on day 1 after plating, recovers to the initial value on day 2 and 3 and afterwards declines exponentially to a low and constant activity level. Ethanol added to the cell culture at a time when low constant activity is reached, reactivates the gamma-glutamyl transpeptidase to 30% of the initial value. PMID- 2565772 TI - Control of reactivation and microtubule sliding by calcium, strontium, and barium in detergent-extracted macrocilia of Beroe. AB - Macrocilia of the ctenophore Beroe are activated to beat continuously in the normal direction by membrane-mediated Ca2+ influx (Tamm: Journal of Comparative Physiology [A] 163:23-31, 1988a). Using saponin or Brij-58 permeabilized models of macrocilia, we show that ATP-reactivation of beating requires microM levels of free Ca2+, Ba2+, or Sr2+. Isolated macrocilia beat initially in reactivation solution (RS) containing Ca2+, Ba2+, or Sr2+ and then undergo microtubule sliding disintegration without added proteases. Addition of protease inhibitors to RS + 10(-5) M Ca2+ prevents sliding disruption. Pretreatment in wash solution (containing 1 mM EGTA) without protease inhibitors, followed by RS + 10(-5) M Ca2+ with protease inhibitors results in extensive sliding disintegration. However, treatment in wash solution followed by RS + protease inhibitors does not induce sliding. Therefore, Ca2+ is not required for proteolysis by endogenous proteases, but is necessary for sliding disintegration. Local iontophoretic application of Ca2+, Ba2+, or Sr2+ to permeabilized macrocilia in RS lacking these cations triggers motility and/or sliding disintegration. Extrusion of microtubules occurs from the tip or the base, depending on whether or not the macrocilium remains attached to its large actin bundle. Thin sheets of microtubules telescope out initially, due to synchronized sliding of subsets of doublet microtubules from parallel rows of axonemes. Macrocilia are one of the first examples of ATP-induced microtubule sliding which retains Ca2+ sensitivity. In addition, the finding that Ba2+ and Sr2+ also trigger active sliding provides an additional method for investigating the control of dynein-powered microtubule movements. PMID- 2565773 TI - Changes in serum concentrations of taurine and other amino acids in clinical antihypertensive exercise therapy. AB - Thirty-one Japanese with essential hypertension were divided into training (n = 21) and non-training (n = 10) groups. Physical training of 10 weeks was instituted after 4 or more weeks of observation, and changes in blood pressure and serum concentrations of taurine and other amino acids were investigated. The workload in physical training was predetermined by the submaximal multistage graded exercise test on a bicycle ergometer, and the blood lactate threshold which reflects approximately 40-60% of maximal oxygen uptake was chosen. The hypertensive patients underwent bicycle ergometer training for 60 minutes, three times a week for 10 weeks. Blood pressures were significantly decreased by 14.8/6.6 mmHg in systole/diastole in the training group, but not in the non training group. Serum concentrations of taurine and cystine were increased significantly by 26% and 287%, in the training group. Increase in serum asparagine (11%), histidine (6%) and lysine (7%) concentrations was also significant, only in the training group. Plasma norepinephrine level and whole blood and plasma volumes were significantly reduced. The change in serum taurine level was significantly negatively correlated with the change in plasma norepinephrine. In addition there was a significant positive correlation between the change in plasma norepinephrine and the change in diastolic blood pressure in the training group. Based on these results, the increase in serum taurine which is known for its antihypertensive activity could contribute, at least in part through the reduction in plasma norepinephrine level, to the antihypertensive effect of exercise. PMID- 2565774 TI - Dopamine receptor blocking action of a dibenzothiepin derivative isofloxythepin in rats. AB - 1. Subcutaneous injection of isofloxythepin or haloperidol inhibited exploratory behaviour, methamphetamine (3 mg/kg)-induced hyperactivity and bromocriptine (15 mg/kg)-induced yawning, and also elicited catalepsy. 2. Isofloxythepin and haloperidol increased concentrations of dihydroxyphenylacetic acid in the striatum and elevated serum prolactin levels. 3. The results suggest that isofloxythepin, as well as haloperidol, blocks the action of the dopamine D2 receptors in the striatum, nucleus accumbens and pituitary. PMID- 2565775 TI - Glucose intolerance in uremic patients: the relative contributions of impaired beta-cell function and insulin resistance. AB - Glucose tolerance and tissue sensitivity to insulin were examined in 19 renal failure patients on chronic regular hemodialysis (group U) and in 6 matched control subjects with normal renal function (group A). Based on glucose tolerance as assessed by an oral glucose tolerance test (OGTT), glucose tolerance was normal in 5 (group U:N), borderline in 5 (group U:BL) and decreased in 9 uremic subjects (group U:D). Compared with group A the uremics demonstrated significantly (p less than 0.01) impaired insulin sensitivity as assessed by a continuous mixed infusion of somatostatin, insulin and glucose (SIGIT). In addition 19 non-diabetic subjects with normal fasting blood glucose and normal renal function, matching the uremic patients with respect to glucose tolerance as assessed by OGTT, were studied (group B). In group B impairments in both insulin secretion and insulin sensitivity tended to be more pronounced in subjects with decreased OGTT as compared with those with borderline OGTT. In contrast, insulin resistance was present to a similar degree in uremic subjects of group U:N, U:BL and U:D. During SIGIT endogenous insulin, glucagon and growth hormone (GH) were suppressed in both uremic and control subjects. This implies that insulin resistance in uremia is most likely not due to hyperglucagonemia or abnormal GH metabolism. During OGTT subjects of group U:N had significantly higher insulin response than subjects of group U:BL (p less than 0.02) and group U:D (p less than 0.01). Insulinogenic index was significantly higher in group U:N than in group U:BL (p less than 0.02) and group U:D (p = 0.01) and was higher in group U:BL than in group U:D (p less than 0.02).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565769 TI - Effects of long-term administration of antidepressants and neuroleptics on receptors in the central nervous system. AB - 1. A review of the effects of long-term administration of antidepressants and neuroleptics on receptors in the central nervous system is presented. 2. The effects of antidepressants on adenylate cyclase activity and on receptor binding in brain tissue are discussed. Effects on a variety of receptor types are considered. 3. The utilization of electrophysiological, behavioral, and neurochemical studies to assess receptor function after chronic antidepressant administration is discussed, as is the use of peripheral receptor estimations in clinical studies. 4. Animal studies on the actions of chronic administration of neuroleptics on pre- and postsynaptic dopamine receptors are reviewed. Effects of these drugs on dopamine receptors in humans are considered from the following perspectives: postmortem and in vivo binding studies in schizophrenia, tardive dyskinesia, and central versus peripheral receptor estimation. PMID- 2565776 TI - Hemodynamic effects of continuous infusion of AR-C 239 in left ventricular failure. AB - AR-C 239 is a new specific alpha 1 antagonist drug with an action similar to that of prazosin. In dogs it appears to be specific for the alpha 1 adrenoreceptor. AR C 239 was tested in 11 patients (49 +/- 11 years old) with left heart failure, who had not received any previous treatment. The drug was infused intravenously with a stepwise increase in dose, producing three stable plasma concentration plateaus: 10 +/- 2, 51 +/- 3, and 138 +/- 55 ng/ml. Hemodynamic data were collected at time T0 before drug infusion, and half-way through each infusion. Blood pressures and cardiac output were recorded using a Millar microtip manometer (left ventricle), a Swan-Ganz catheter (pulmonary artery), and a femoral catheter, connected to a Syscomoram computer system which calculated the following parameters: work, resistances, tension-time index and contractility indices. Left ventricular ejection fraction and volumes were obtained from cineangiograms performed 30 min before drug infusion and at the end of the third infusion plateau. AR-C 239 produces a dose-dependent fall in systemic blood pressure (r = 0.539, n = 33, p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565777 TI - Rapid DNA extraction and sensitive alkaline blotting protocol: application for detection of gene rearrangement and amplification for clinical molecular diagnosis. AB - A simple and sensitive DNA extraction/Southern blotting method is described for the routine detection of gene rearrangement and amplification in DNA samples. Significant features of the procedure are: (1) sequential digestion of RNA and protein in the same DNA buffer; and (2) use of a modified alkaline transfer protocol which increases sensitivity and decreases the autoradiographic exposure period. The method lends itself to handling multiple samples containing minimal amounts of starting material, requires equipment commonly found in a clinical laboratory, and yields results within 36 h of specimen receipt. Accordingly, these procedures may find utility in the establishment of molecular diagnostics in a conventional clinical setting. PMID- 2565778 TI - Restriction fragment length polymorphism of HLA and non-HLA genes in DR3/4 heterozygous Danish insulin-dependent diabetic patients and healthy individuals: reassessment of the influence of alpha DX and insulin-linked polymorphic loci, and new splits of DQw3 haplotypes. AB - We have investigated restriction fragment length polymorphism (RFLP) of HLA and non-HLA regions of the genome in the homogeneous Danish population. Insulin dependent diabetes mellitus (IDDM) patients and healthy individuals were selected for being HLA-DR 3/4 heterozygous to evaluate the influence of genes other than DR on disease susceptibility. Five different probes were used: HLA alpha and beta DQ (chromosome 6), the Ins 310 genomic fragment which detects a polymorphic region 5' to the insulin gene (chromosome 11), and cDNA for the constant regions of the T cell receptor alpha and beta genes (chromosomes 14 and 7). Fifteen cells homozygous for the HLA-D region were used to obtain reference DNA patterns. This allowed us to describe four splits among the HLA-DQw3 haplotypes (DQw3.1 to DQw3.4). The two new haplotypes DQw3.3 and DQw3.4 do not code for the TA10 serological marker which is found on DQw3.1 positive cells. One-hundred per cent of IDDM patients were typed as DQw3.2 versus 68 per cent for controls (p = 0.003). However, our results do not indicate a role for the Ins 310 or for the alpha DX locus region in IDDM susceptibility, in contrast to previous reports by others. The restriction enzymes that we have used did not reveal significant differences between DNA patterns of patients and controls with probes for the constant region of the T cell receptor genes. PMID- 2565780 TI - Seasonal changes in the red blood cell system in the European bison, Bison bonasus L. AB - 1. In 195 European bisons divided into four groups (Group 1, 0-3 year-old males; Group 2, 0-3 year-old females; Group 3, mature bulls, over 3 years old; Group 4, mature cows, over 3 years old) seasonal changes in red blood cell number and diameter, haemoglobin level, haematocrit value, index F, MCH, MCHC and MCV were studied. 2. Seasonal cyclicity was found only in red blood cell diameters in all groups. 3. In young males the cyclicity was found in MCHC and in MCV only. 4. In young females the cyclicity was found in Hb and Hct values, in MCH and in MCV. 5. In mature bulls cyclicity was found only in RBC diameter and in MCV. 6. In mature cows the cyclicity was found in index F, MCH and in MCHC values. 7. Seven out of 14 acrophases of cyclic indices occurred just before autumnal equinox and three before vernal equinox. PMID- 2565779 TI - Basal and tolbutamide-induced plasma somatostatin in healthy subjects and in patients with diabetes and impaired glucose tolerance. AB - Peripheral levels of basal and tolbutamide-induced somatostatin have been measured in patients with diabetes or impaired glucose tolerance (IGT) and compared with those in normal individuals. Basal somatostatin was significantly higher in patients with Type 1 diabetes than in age-matched control subjects. This increase was most pronounced at diagnosis, and appeared to be related to metabolic control in insulin-treated patients. No increase was noted in patients with Type 2 diabetes or with IGT. Intravenous bolus injection of tolbutamide enhanced peripheral somatostatin levels in healthy volunteers in a biphasic manner. Patients with IGT also exhibited a biphasic response but the amplitude of the first phase was higher. No secretory response was detected in 27/29 Type 1 diabetic patients at diagnosis; a somatostatin response to tolbutamide became detectable again in Type 1 patients with normalization of their basal somatostatin levels but was then paradoxically related to poor blood glucose control. In Type 2 diabetes, basal somatostatin levels were similar to age matched control subjects, but decreased upon intravenous tolbutamide administration. PMID- 2565781 TI - Tissue specific differences in the fatty acid composition of the marmoset monkey (Callithrix jacchus). AB - 1. The possible relationship between the fatty acid composition of the adipose tissue of the marmoset monkey (Callithrix jacchus) and its dietary lipid intake was examined after a long-term feeding trial. 2. Only the proportions of stearic and linoleic acid in the adipose tissue were similar to those in the diet. The proportions of all other saturated and unsaturated fatty acid components were significantly different thus greatly reducing the potential value of this relationship in the marmoset. 3. The phospholipid fatty acid profiles of cardiac and skeletal muscle, aortae, kidney, liver, lung and brain were also compared to those of the plasma, platelets or red blood cells. 4. With the exception of the liver where the fatty acid profile is similar to that of the plasma, no clear relationships were found between the fatty acid profiles of these tissues and the components of the blood. 5. The fatty acid composition of skeletal muscle was very similar to that of the cardiac muscle of the marmoset, suggesting that muscle biopsy might be useful as an index of cardiac muscle composition. PMID- 2565782 TI - Cathepsin B, D and H activities in muscles of chicks of fast and slow growing strains: effect of age and diet. AB - 1. Activities of cathepsins B, D and H were measured in leg and breast muscles of fast growing (broiler) and slow growing (layer) chicks at eight time intervals between 1 and 29 days of age. 2. These enzyme activities were also measured in muscles from fast and slow growing chicks given a low protein (125 g/kg crude protein) diet between the ages of 17 and 24 days. 3. Activities of none of these cathepsins differed greatly between muscle type or strain of chick. However in both strains of chick cathepsin D and H in muscles significantly decreased with increasing age (muscle size) of the chick. Cathepsin D activity also increased when muscle proteolytic rates were increased by feeding a low protein diet. This latter effect was significant only in the muscles of fast growing chicks. 4. The results suggest that lysosomal proteases are not responsible for the differences in muscle protein degradation and growth between fast and slow growing strains of chicks, or between muscle types in the chick. PMID- 2565783 TI - Enhanced release of cholecystokinin by dietary amino acids in chicks (Gallus domesticus). AB - 1. The effect of dietary amino acids and protein on cholecystokinin (CCK) release into plasma was investigated in chicks by feeding a meal through a stomach tube, followed by the CCK determination with specific CCK-8 antibody. 2. The results showed that both isolated soya protein and an amino acid mixture simulating the amino acid composition of the soya protein increased the release of CCK, though to a lesser extent with a delayed response in the former, when added to a protein free diet. 3. Among amino acids added singly to the protein-free diet, phenylalanine was more efficient than arginine and valine, exerting a response almost identical to the complete amino acid mixture. PMID- 2565784 TI - Cardiovascular hemodynamics in the swine following long-term placement of prosthetic grafts post right ventriculotomy. AB - 1. Heart rate, cardiac output and arterial, pulmonary artery, central venous and pulmonary wedge pressures were measured in eight neutered male pigs of an average of 14 months of age and 145.0-231.0 kg body weight. 2. These measurements were made 10-16 months after right ventriculotomy and placement of Gore-Tex PTFE Vascular Grafts. 3. Arithmetic means, standard deviations and coefficients of variation were calculated and the 95% confidence limits for ranges were established. 4. Comparison of derived data with that from published data on swine and on humans indicated normal cardiovascular hemodynamics in this group. 5. In addition, there was no gradient across the patched area. 6. It appears, therefore, that these patches had no adverse effects on the cardiovascular system of growing pigs. PMID- 2565785 TI - Regulation of weight loss in male farm mink. AB - 1. Regulation of weight loss was studied in wild mink (Mustela vision) under farm conditions. 2. Body weight of male minks was already adjusted to a low level before the mating season. 3. Weight loss was a result of changes in energy expenditure and energy intake. 4. This study emphasizes the dynamic nature of seasonal energy regulation. PMID- 2565786 TI - Basal rate of metabolism and temperature regulation in Sorex coronatus and S. minutus (Soricidae: Mammalia). AB - 1. The basal rate of metabolism (BMR), body temperature and thermal conductance of Sorex minutus and Sorex coronatus were studied. Special attention was given to measuring BMR in resting and postabsorptive shrews. 2. Both species exhibit a high BMR, respectively 339 and 289% of the value expected on the basis of body mass. 3. The thermal conductance of both species is similar to the predicted value and their body temperature averages 38.6 and 37.6 degrees C respectively. 4. Available data suggest that a very high BMR may be a common feature within the genus Sorex. PMID- 2565787 TI - Behavioral discrimination between glutamate and the four basic taste substances in mice. AB - 1. Behavioural studies using the conditioned taste aversion (CTA) paradigm in mice showed that aversion conditioned to monosodium L-glutamate (MSG), which elicits a unique taste in humans, did not strongly generalize to any of the four basic taste stimuli, suggesting that mice could behaviourally discriminate between MSG and the four basic taste stimuli. 2. Denervation of bilateral glossopharyngeal nerve significantly increased behavioural similarities (the strength of generalization in the CTA paradigm) between MSG and sodium salts. This was not the case after destruction of the bilateral chorda tympani nerve. 3. These results suggest that taste information of glossopharyngeal nerve plays a more important role in the behavioural discrimination between MSG and the four basic tastes than does that of the chorda tympani nerve. PMID- 2565788 TI - Peripheral neural basis for behavioural discrimination between glutamate and the four basic taste substances in mice. AB - 1. Single chorda tympani fibres sensitive to monosodium L-glutamate (MSG), elicit a unique taste in humans and gave a greater response to NaCl and/or sucrose than to MSG whereas several MSG-sensitive glossopharyngeal fibres responded only slightly if at all to NaCl and sucrose. 2. The across-fibre correlations showed that MSG and NaCl produced similar response patterns in the chorda tympani fibres but different response patterns in the glossopharyngeal fibres. 3. These results suggest that taste information of glossopharyngeal fibres plays a relatively more important role in the qualitative discrimination between MSG and the four basic taste substances than that of chorda tympani fibres. PMID- 2565789 TI - Evaluation of mammary blood flow measurements in lactating goats using the ultrasound Doppler principle. AB - 1. Non-invasive methods were developed for measuring mammary blood flow in lactating goats. 2. A Doppler principle ultrasound device was equipped with an external detector measuring maximal blood velocity (Vmax) and average blood velocity (Vav) was calculated as Vmax/2. Volume flow then depended on determination of the angle of insonation and the cross-sectional area of the milk vein (the caudal superficial epigastric or subcutaneous abdominal vein). 3. Blood velocities were measured on the milk vein of either side of the animal while clamping the pudendal veins manually. Blood velocities ranged from 7-34 cm/sec. 4. The milk vein diameter was measured by means of a slide gauge which, for clearly protruding veins, gave similar results to that measured by ultrasound scanning. In protruding veins the cross-section was circular. In non-protruding veins the cross-section was elliptical and the slide gauge significantly (P less than 0.01) overestimated the cross-sectional area. The milk vein diameter of either side measured in 10 lactating goats was 8.8 +/- 1.1 mm (means +/- SD). 5. Blood flow ranged from 90-675 ml/min in a dry and a high-yielding (3.4 l milk daily) goat, respectively. The reproducibility of the blood flow measurements was 12-16%. 6. It is concluded that the present method may be used for quantitative measurements of mammary blood flow in goats. PMID- 2565790 TI - Changes in the adrenal steroidogenic responsiveness of the mallard duck (Anas platyrhynchos) during early post-natal development. AB - 1. Plasma concentrations of corticosterone (B), aldosterone (Aldo) and deoxycorticosterone (DOC) were measured in mallard ducklings immediately before and after exposure to acute immobilization stress. 2. Except for transient declines in B and DOC between the 4th and 14th days after hatching, the resting concentration of each hormone did not change significantly during post-natal development. 3. The stress-induced in Aldo was maximal at hatching while maximal increases in B and DOC did not occur until one day later. 4. Thereafter the magnitude of the stress-induced increases in the concentrations of all of the hormones decreased steadily and on the 21st and 28th days after hatching only B increased significantly in response to stress. PMID- 2565791 TI - Development of temperature regulation in the common white-toothed shrew, Crocidura russula. AB - 1. Body temperature and oxygen consumption were measured during the first month of postnatal life in two litters of Crocidura russula at four different ambient temperatures. 2. Body temperature in the nest varies from 30.5-35.0 degrees C during the first 10 days; afterwards it becomes more constant (35.5-37.0 degrees C). 3. First homoiothermic reactions occur on the 2nd day of life and become effective on the 4th day. 4. In the case of undernourished shrews, torpor is already developed on the 2nd day. 5. After the 7th day torpor is the obligatory reaction in cooling experiments. 6. This discontinuous development of temperature regulation is interpreted as typical for shrews capable of torpor. PMID- 2565792 TI - Relationships between renal hemodynamics and plasma levels of arginine vasotocin and mesotocin during hemorrhage in the domestic fowl (Gallus domesticus). AB - 1. Renal tissue blood flow (renal perfusion) and plasma levels of arginine vasotocin (AVT) and mesotocin (MT) were measured in anesthetized chickens before and during hemorrhage. 2. Renal perfusion did not decrease (P less than 0.05) until nearly 50% of the blood volume had been removed. The decrease in renal perfusion was not related to arterial blood pressure but was concomitant with an increase (P less than 0.05) in plasma AVT levels. 3. Renal perfusion during hemorrhage was positively correlated with plasma MT levels by the regression equation: renal perfusion = 0.091 (MT)-1.1459 which was highly significant (P less than 0.001, r2 = 0.95). 4. The results of this study suggest that MT as well as AVT may participate in regulating blood flow in the avian kidney. PMID- 2565793 TI - Properties of barnacle photoreceptor cells in culture. AB - 1. Properties of median photoreceptor cells in cultured ocelli from the giant barnacle (Balanus nubilus) were compared in isolated ocelli, ocelli maintained with the supraesophageal ganglion, and fresh ocelli. 2. Cultured photoreceptor cells exhibited slight deterioration after 2-4 weeks. Cell bodies maintained their structure but apparently lost some dendrites. Electron micrographs revealed fewer rhabdomeres. Axons did not degenerate. 3. Intracellularly recorded responses to light in both cultured preparations were qualitatively normal with a small decrease in sensitivity and increase in input resistance. The waveforms of the light responses were normal. 4. The characteristic shadow reflex was maintained after 6 weeks. PMID- 2565794 TI - Centrifugal decrement in diameter of myelinated afferent fibres innervating frog taste organ. AB - 1. Regional changes in the diameter of single myelinated afferent nerve fibres innervating the taste disc of the fungiform papillae on the bullfrog tongue were investigated morphologically and functionally. 2. The diameter of myelinated afferents in the medial lingual branch of the glossopharyngeal nerve averaged 8.4 microns at the proximal end of the tongue and gradually decreased at the rate of 0.8 micron/cm length of the fibres as they ran in the apical direction of the tongue. 3. The conduction velocity of single myelinated afferent fibres within the tongue decreased gradually as they ran peripherally. 4. Electrophysiological inspection of neural connections between the fungiform papillae suggests that a gradual centrifugal decrease in the diameter of a single myelinated afferent fibre is not due to multiple bifurcations of the fibre at various sites within the tongue, but due to a natural gradual decrease in the thickness of the myelin sheath and the diameter of axon. PMID- 2565795 TI - Expression of myosin in atrial areas of the bovine myocardium. AB - 1. A comparison of myosins from defined areas of the bovine atrial myocardium was performed by measuring Ca2+-ATPase activity and electrophoretic separation of myosin light chains. 2. Some areas of atrial myocardium contained myosin with slightly higher ATPase activity than others. 3. There were also clear differences in the amount of one ventricular light chain of myosin in defined regions of atrial myocardium. 4. No close relationship existed between the expression of ventricular and atrial myosin light chains and myosin ATPase activity. PMID- 2565796 TI - Biochemical correlates of the structural allometry and site-specific properties of mammalian adipose tissue. AB - 1. The maximum activities of the glycolytic enzymes hexokinase (HK) and phosphofructokinase (PFK) were measured in defatted homogenates of adipose tissue from nine homologous depots of 57 wild and captive mammals belonging to 17 species and eight orders and differing in body mass by six orders of magnitude. 2. Site-specific differences in the enzyme activities were similar in all terrestrial species and were not consistently related to adipocyte volume. 3. The specimen-mean maximum activities of HK and PFK did not correlate with body mass, body composition or natural diet. 4. When specimens of different body composition and body mass were compared, glycolytic enzyme activity per adipocyte was directly proportional to adipocyte volume. 5. Site-specific differences in collagen content of adipose tissue did not correspond to those adipocyte volume. When homologous depots of different specimens were compared, the collagen content of adipose tissue was directly proportional to body mass. 6. Adipose tissue of large cetaceans contains more collagen than predicted from the allometric equations fitted to the data from terrestrial mammals. 7. Neither the scaling of the collagen content with body mass nor the site-specific differences in its abundance are consistent with a role as protection or support for adjacent tissues. 8. There are consistent site-specific differences in the extracellular components of adipose tissue as well as in the structure and metabolism of the adipocytes. 9. Adipose tissue differs from most other tissues in that its maximum metabolic capacities do not scale to body mass. 10. Adjustment of the biochemical activity of adipose tissue to changes in body mass and body composition must depend upon neural and endocrine controls, not upon intrinsic differences in its metabolic capabilities. PMID- 2565797 TI - Taxol, cytochalasin B and colchicine effects on fibroblast migration and contraction: a role in glaucoma filtration surgery? AB - Migration and contraction are fibroblast activities in scar tissue formation which may contribute towards the failure of trabeculectomies. We attempted to modify the migration and contraction of Tenon's capsule fibroblasts in vitro using the drugs taxol, cytochalasin B and colchicine. Migration assays were conducted in 48-well micro-chemotaxis chambers, using rabbit aqueous humour which has been previously identified as a powerful chemoattractant for Tenon's fibroblasts, and fibronectin as the stimuli for migration. All three drugs inhibited the migration of fibroblasts to both fibronectin and aqueous humour, with maximal activity seen at 10(-6)M (taxol), 10(-5)M (cytochalasin B), and 10( 3) M (colchicine). In a single cell contractile assay in which contraction of whole fibroblasts was initiated with exogenous adenosine triphosphate (ATP), taxol and colchicine showed anticontractile activity, maximal at 10(-5)M and 10( 3)M respectively, but cytochalasin B was ineffective. The significance of the activities of these drugs in relation to their proposed use for improving the success of trabeculectomies is discussed. PMID- 2565798 TI - Human retrovirology. Facts and concepts. PMID- 2565799 TI - Male infertility related to 5-ASA enemas. PMID- 2565800 TI - [Enlistment of assistant physicians without proof of qualification for the rescue service]. PMID- 2565801 TI - Canine testicular tumors in descended and cryptorchid testes. AB - A comparative study of canine testicular tumors in descended and cryptorchid testes was performed in order to associate pathologic features with clinical signs. Observation on 80 canine testicular tumors revealed the following distribution: in undescended testes, 4 tumors were classified as Seminomas (mean age: 8.8 +/- 3 years) and 14 as Sertoli cell tumors (8.7 +/- 1.7 years); in descended testes, 21 tumors were classified as seminomas (8.8 +/- 3 years), 13 as Sertoli cell tumors (9.8 +/- 1.8 years), 22 as Leydig cell tumors (11.5 +/- 2 years), 5 multiple primary tumors and 1 as an immunoblastic lymphoma. Histological features were studied and correlated with other clinical parameters: feminization manifestations, prostatic disease, perineal hernia and perianal gland tumor. PMID- 2565803 TI - Postnatal development of tyrosine-hydroxylase-immunoreactive cells in the rat retina. Morphology and distribution. AB - The appearance and development of tyrosine-hydroxylase-immunoreactive cells were studied using whole mounts of postnatal rat retinas. The two classes of cells described in the adult retina were observed in the young rat. Small unipolar cells appeared as soon as the day of birth in the superior retina and large stellate cells by postnatal day (PND) 7. By PND 10, their number reached that of stellate cells of the adult-retina. It is hypothesized that small unipolar cells are transformed into large stellate cells as they mature. Several arguments are put forward to support this conclusion, the most important of which is the appearance of peripheral crescents containing a high density of small cells which disappear proportionately as the number of large stellate cells increases. The development of the vasculature was also studied. By PND 7, it was more developed in the temporal superior quadrant, in the region where the first stellate cells appeared. PMID- 2565804 TI - Ethanol in clinically relevant concentrations enhances expression of oligodendroglial differentiation but has no effect on astrocytic differentiation or DNA synthesis in primary cultures. AB - Primary cultures of cerebral glia derived from newborn rat brain were utilized to evaluate the effects of ethanol on DNA synthesis and cellular differentiation. Glutamine synthetase was employed as a marker for astrocytic differentiation and 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNP), for oligodendroglial differentiation. Concentrations of ethanol of 17-86 mM, i.e., a concentration range comparable to that observed in humans, were utilized. No effect of ethanol on DNA synthesis was observed, despite the use of synchronized cultures. Similarly, no effect of ethanol on the developmental increase of glutamine synthetase activity was seen, despite the use of astrocytes purified by the selective detachment technique and a prolonged duration of exposure to ethanol (17 days). In contrast, however, a striking enhancement of CNP activity was demonstrable in both mixed glial cultures and in oligodendroglia purified by the selective detachment technique. In the latter cells, the stimulatory effect of ethanol was evident within 9 days of exposure, and after 17 days of exposure, ethanol-treated cells exhibited a two-fold higher CNP activity than did control cells. This peak effect was observed at an ethanol concentration of only 17 mM. Thus, these data indicate that (1) clinically relevant concentrations of ethanol have no effect on either DNA synthesis or on at least one expression of astrocytic differentiation in glial primary cultures, but (2) these concentrations exert a striking enhancement of CNP activity, a marker of oligodendroglial differentiation. The findings have implications both for the effects of ethanol on the developing nervous system and the regulation of oligodendroglial differentiation. PMID- 2565805 TI - Impact of chromium and tin on a nitrogen-fixing cyanobacterium Anabaena doliolum: interaction with bivalent cations. AB - The toxicity of chromium and tin on growth, uptake of NO3- and NH4+, nitrate reductase and glutamine synthetase activity of Anabaena doliolum, and its interaction with bivalent cations, viz. Ca2+, Mg2+, Mn2+, Ni2+, Co2+, and Zn2+, has been studied. Some interacting cations, viz. Ca, Mg, and Mn, substantially antagonized the toxic effects of chromium and tin with reference to growth and nutrient (NO3- and NH4+) uptake in the hierarchical sequence Ca greater than Mg greater than Mn, whereas the sequence of hierarchy was Mn greater than Mg greater than Ca for nitrate reductase and glutamine synthetase activity of A. doliolum. A synergistically inhibitory pattern of interaction was noted for all the parameters, viz. growth, uptake of NO3- and NH4+, nitrate reductase and glutamine synthetase activity of A. doliolum, when Ni, Co, and Zn were used in combination with test metals in the growth medium. These bivalent cations followed the synergistic inhibition sequence Ni greater than Co greater than Zn and potentiated the toxicity of test metals in the N2-fixing cyanobacterium under study. PMID- 2565802 TI - Histamine2 (H2)-receptor antagonists in the treatment of urticaria. AB - Urticaria may develop in response to a number of stimuli such as allergic reactions, drugs, cold, pressure, stings and, most interestingly, neuropsychological upheavals. Classical treatment has utilised H1-receptor antagonists, in view of the fact that histamine released from local mast cells acts on H1-receptors on the vasculature and participates in the pathophysiology of this syndrome. More recently, H2-receptor antagonists have also been tried, alone or in combination, with encouraging results. The question still remains why H2-receptor antagonists should have any beneficial effect since H2-receptors are mostly present on exocrine cells and on T-suppressor lymphocytes, where they are stimulatory, or mast cells, where they are auto-inhibitory. Possible explanations may include the ratio of H1- to H2-receptors on local vasculature and the effect of H2-antagonists on responses elicited through nervous system activity via cholinergic or neuropeptidergic neurons. Finally, evidence is presented that certain tricyclic H-receptor antagonists may have powerful inhibitory effects on secretion from both peripheral and central nervous system mast cells, as well as from neurons. The possible role of H3-receptors in this process is also discussed. At present, the available evidence does not justify the routine use of H2-antagonists in the treatment of urticaria. PMID- 2565806 TI - Effect of endogenous opioid peptides on TRH release from rat stomach in vitro. AB - The effect of endogenous opioid peptides (beta-endorphin, leucine-enkephalin, dynorphin 1-13 on the release of thyrotropin releasing hormone (TRH) from the rat stomach in vitro was studied. The rat stomach was incubated in the medium 199 with 1.0 mg/ml of bacitracin (pH 7.4) (medium) and the amount of TRH released into the medium was measured by radioimmunoassay. The release of TRH from the rat stomach was inhibited significantly in a dose-related manner with the addition of endogenous opioid peptides, but not affected with naloxone. However, the inhibitory effect of endogenous opioid peptides on TRH release from the rat stomach was prevented by the addition of naloxone. These findings suggest that endogenous opioid peptides inhibit TRH release from the rat stomach in vitro. PMID- 2565807 TI - A chimeric EGF-R-neu proto-oncogene allows EGF to regulate neu tyrosine kinase and cell transformation. AB - The neu oncogene, characterized by Weinberg and colleagues, is a transforming gene found in ethylnitrosourea-induced rat neuro/glioblastomas; its human proto oncogene homologue has been termed erbB2 or HER2 because of its close homology with the epidermal growth factor receptor (EGF-R) gene (c-erbB1). Expression of the rat neu oncogene is sufficient for transformation of mouse NIH 3T3 fibroblasts in culture and for the development of mammary carcinomas in transgenic mice, but the neu proto-oncogene has not been associated with cell transformation. We constructed a vector for expression of a chimeric cDNA and hybrid protein consisting of the EGF-R extracellular, transmembrane and protein kinase C-substrate domains linked to the intracellular tyrosine kinase and carboxyl terminal domain of the rat neu cDNA. Upon transfection with the construct, NIH 3T3 cells gave rise to EGF-R antigen-positive cell clones with varying amounts of specific EGF binding. Immunofluorescence and immunoprecipitation using neu- and EGF-receptor specific antibodies demonstrated a correctly oriented and positioned chimeric EGF-R-neu protein of the expected apparent mol. wt on the surface of these cells. EGF or TGF alpha induced tyrosine phosphorylation of the chimeric receptor protein, stimulated DNA synthesis of EGF R-neu expressing cells and led to a transformed cell morphology and growth in soft agar. In contrast, the neu proto-oncogene did not show kinase activity or transforming properties when expressed at similar levels in NIH 3T3 cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565808 TI - HER2 cytoplasmic domain generates normal mitogenic and transforming signals in a chimeric receptor. AB - We have investigated the biological function of an unidentified human growth factor, the ligand of the putative HER2 receptor, by characterizing the signalling properties of its receptor. HER2 (or c-erbB-2), the human homolog of the rat neu proto-oncogene, encodes a transmembrane glycoprotein of the tyrosine kinase family that appears to play an important role in human breast carcinoma. Since a potential ligand for HER2 has not yet been identified, it has been difficult to analyze the biochemical properties and biological function of this cell surface protein. For this reason, we replaced the HER2 extracellular domain with the closely related ligand binding domain sequences of the epidermal growth factor (EGF) receptor, and examined the ligand-induced biological signalling potential of this chimeric HER1-2 protein. This HER1-2 receptor is targetted to the cell surface of transfected NIH 3T3 cells, forms high and low affinity binding sites, and generates normal mitogenic and cell transforming signals upon interaction with EGF or TGF alpha. The constitutive activation of wild-type HER2 in transfected NIH 3T3 cells suggests the possibility that these cells synthesize the as yet unidentified HER2 ligand and activate HER2 by an autocrine mechanism. PMID- 2565809 TI - The homeotic gene Sex Combs Reduced of Drosophila: gene structure and embryonic expression. AB - The homeotic gene Sex Combs Reduced (Scr) of Drosophila is required during embryogenesis for labial and first thoracic segment development. We define the Scr gene structure, showing that the major embryonic transcript is proximal to the fushi tarazu gene, and report the sequence of the transcript, which encodes a 413-amino acid, homeodomain-containing protein. We describe Scr protein distribution throughout embryogenesis. Expression begins at gastrulation and is eventually apparent in three tissues, epidermis, nervous system and visceral mesoderm, though there are clear contrasts in the domains of expression in these three tissues. PMID- 2565811 TI - Metabolism of cysteinyl leukotrienes in non-recirculating rat liver perfusion. Hepatocyte heterogeneity in uptake and biliary excretion. AB - 1. The uptake, metabolism and biliary excretion of the cysteinyl leukotrienes LTC4, LTD4 and LTE4, were studied in a non-recirculating rat liver perfusion system at constant flow in both antegrade (from the portal to the caval vein) and retrograde (from the caval to the portal vein) perfusion directions. During a 5 min infusion of [3H]LTC4, [3H]LTD4 and [3H]LTE4 (10 nmol/l each) in antegrade perfusions single-pass extractions of radioactivity from the perfusate were 66%, 81% and 83%, respectively. Corresponding values for LTC4 and LTD4 in retrograde perfusions were 83% and 93%, respectively, indicating a more efficient uptake of cysteinyl leukotrienes in retrograde than in antegrade perfusions. The concentrations of unmetabolized leukotrienes in the effluent perfusate were 8-12% in antegrade and 2-4% in retrograde perfusions. [14C]Taurocholate extraction from the perfusate was inhibited by LTC4 by only 3%, suggesting that an opening of portal-venous/hepatic-venous shunts does not explain the effects of perfusion direction on hepatic LTC4 uptake. 2. Following infusion of [3H]LTC4 and [3H]LTD4, in the antegrade perfusion direction, about 80% and 87%, respectively, of the radiolabel taken up by the liver was excreted into bile. In retrograde perfusions, however, only 40% and 57%, respectively, was excreted into bile and the remainder was slowly redistributed into the perfusate, indicating that leukotrienes were taken up into a hepatic compartment with less effective biliary elimination or converted to metabolites escaping biliary excretion. The metabolite pattern found in bile was not affected by the direction of perfusion. Biliary products of LTC4 were polar metabolites (31-38%), LTD4 (27-30%), LTE4 (about 1%) and N-acetyl-LTE4 (3-4%) in addition to unmodified LTC4 (17-18%). 3. LTC4 was identified as a major metabolite of [3H]LTD4 in bile, amounting to about 20% of the total radioactivity excreted into bile. This is probably due to a gamma-glutamyltransferase-catalyzed glutamyl transfer from glutathione in the biliary compartment, as demonstrated in in vitro experiments. The presence of sinusoidal gamma-glutamyltransferase activity in perfused rat liver was shown in experiments on the hydrolysis of infused gamma-glutamyl-p-nitroanilide. 90% inhibition of this enzyme activity by AT-125 did not affect the metabolism of LTC4. 4. When [3H]LTE4 was infused in the antegrade perfusion direction, biliary metabolites comprised N-acetyl-LTE4 (24%) and polar components (60%).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2565810 TI - Hox-7, a mouse homeobox gene with a novel pattern of expression during embryogenesis. AB - A new mouse Hox locus, Hox-7, is defined on chromosome 5 by a gene homologous to the Drosophila gene msh, which contains a homeobox sequence distantly related to that of Antennapedia. By in situ hybridization, expression of Hox-7 is detected in the neural fold of embryos, and also in cephalic neural crest. In addition, expression takes place in the developing valves of the embryonic heart. Mandibular and hyoid arches are strongly labelled, expression becoming restricted to the most distal part of mouth and face processes as development proceeds. Intense labelling is also observed in developing limb buds, in the distal region which has been shown to be essential for limb morphogenesis. The pronounced accumulation and regional localization of Hox-7 transcripts in mandibular and limb processes point to a specific morphogenetic role for this mouse homeobox gene. PMID- 2565812 TI - Energy requirement and kinetics of transport of poly(A)-free histone mRNA compared to poly(A)-rich mRNA from isolated L-cell nuclei. AB - ATP-promoted efflux of poly(A)-rich RNA from isolated nuclei of prelabeled mouse lymphoma L5178y cells has an activation energy of 51.5 kJ/mol, similar to that found for the nuclear envelope nucleoside triphosphatase (48.1 kJ/mol) assumed to be involved in mediating nucleocytoplasmic transport of at least some RNA. Here we show that efflux of two specific poly(A)-rich mRNAs (actin and beta-tubulin) from isolated L-cell nuclei is almost totally dependent on the presence of ATP, while efflux of poly(A)-free histone mRNA (H4, H2B, and H1) also occurs to a marked extent in the absence of this nucleotide. Measurements of temperature dependence of transport rate revealed an activation energy of 56.1 kJ/mol for actin mRNA, while the activation energy for histone-H4-mRNA efflux was in the same range as that found for ATP-induced release of RNA from demembranated nuclei (about 15-20 kJ/mol). Addition of nonhydrolyzable nucleotide analogs of ATP to the in vitro system used for measurement of RNA transport did not result in release of nonhistone mRNA (actin), but enhanced the efflux of H4 mRNA to approximately the same extent as ATP. Although not absolutely required, addition of ATP stimulated the rate of export of histone mRNA about twofold. Only the poly(A)-rich RNA, but not the poly(A)-free RNA, released from isolated nuclei was found to compete with poly(A) for the nuclear envelope mRNA-binding site, indicating the mechanism of transport for both RNA classes to be distinct. Export of both nonhistone and histone mRNA was found to be inhibited by a monoclonal antibody against a p60 nuclear-pore-complex antigen. This antibody had no effect on the nucleoside triphosphatase, mediating transport of poly(A)-rich mRNA. PMID- 2565813 TI - Reliable five-minute test strip method for identification of Streptococcus pyogenes. AB - A novel and rapid method (Strep Strip, Lab M) for identification of Streptococcus pyogenes was evaluated. The method combines the established test for pyroglutamyl aminopeptidase (PYR) with a rapid chromogenic test for beta-glucosidase on a paper strip. The test was evaluated with 274 clinical isolates and 237 culture collection isolates of beta-haemolytic streptococci. Streptococcus pyogenes was identified with 100% specificity. Six isolates identified as Lancefield group A and which might therefore be assumed to be Streptococcus pyogenes were shown by the test strip method not to be this species. The beta-glucosidase test on the test strip allowed differentiation between enterococci and Streptococcus pyogenes (both PYR positive) in all cases. The test strip method represents an economical and accurate method for identification of Streptococcus pyogenes in clinical material. PMID- 2565814 TI - Detection of the multidrug resistant phenotype in human tumours by monoclonal antibodies and the streptavidin-biotinylated phycoerythrin complex method. AB - The aim of this study was to find out whether the membrane glycoprotein P-170 can be detected in human tumours with both acquired and intrinsic resistance to chemotherapeutic agents using monoclonal antibodies (265/F4 and C219) and the streptavidin-biotinylated phycoerythrin complex method. Pretreated leukaemia cells and untreated lung and ovarian carcinomas were analysed. Two plasmacytomas and one leukaemia expressed high levels of P-glycoprotein, whereas two leukaemias showed moderate, and three leukaemias no expression of this protein. The intrinsic resistance was analysed with a panel of four human epidermoid lung cancer xenografts grown in nude mice. The expression of P-glycoprotein could be correlated with the degree of resistance. In addition, one out of five ovarian carcinomas revealed a high level of P-glycoprotein. PMID- 2565815 TI - Effects of classical and atypical antipsychotic drugs on isolation-induced aggression in male mice. AB - A series of classical, atypical and putative antipsychotic drugs were compared for their ability to inhibit isolation-induced intraspecies aggression with affinity for D-2 dopamine receptors and induction of akinesia. The majority of drugs tested significantly inhibited aggressive behavior only after doses that greatly decreased the ability of mice to move. Even though akinesia seemed to account for inhibition of aggression there was no apparent correlation with binding to striatal D-2 receptors. PMID- 2565816 TI - CY 208-243, a novel dopamine D-1 receptor agonist, fails to modify dopamine release in freely moving rats. AB - CY 208-243, a novel D-1 agonist structurally unrelated to other D-1 agonists, at doses which elicited behavioural stimulation with locomotion, sniffing and grooming, failed to modify the release and metabolism of dopamine (DA) in the nucleus accumbens and in the dorsal caudate of freely moving rats, as estimated by transcerebral dialysis. CY 208-243 prevented the increase of DA release and metabolism elicited by the specific D-1 antagonist, SCH 23390, but not by the specific D-2 antagonist, sulpiride. The results support the conclusion that CY 208-243 is an effective and specific D-1 agonist in vivo. PMID- 2565817 TI - Biochemical and functional studies on EMD 49,980: a potent, selectively presynaptic D-2 dopamine agonist with actions on serotonin systems. AB - EMD 49,980 proved to be a potent and selectively presynaptic D-2 dopamine receptor agonist in biochemical studies with rats. Thus, the gamma-butyrolactone induced accumulation of dihydroxyphenylalanine, used as a presynaptic model, was antagonized with ED50 values of 0.29 and 0.09 mumol/kg in striatum and t. olfactorium, respectively, with high maximal effects. In contrast, striatal acetylcholine concentrations, reflecting actions at normosensitive postsynaptic D 2 receptors, were only moderately increased by about 30% over a dose range of 2.3 68 mumol/kg. In rats with unilateral nigrostriatal lesions, EMD 49,980 induced long-lasting contralateral turning, indicative of agonistic actions at denervated postsynaptic D-2 receptors. In addition, EMD 49,980 potently inhibited serotonin (5-HT) uptake in vitro and in vivo. Binding studies confirmed D-2 activity in the nM range but, similarly potent effects were observed at 5-HT1A binding sites. Measurement of 5-hydroxytryptophan (5-HTP) accumulation in the n. raphe revealed that, in vivo, the net effect of EMD 49,980 on 5-HT systems is an agonistic one. Control experiments indicate that inhibition of 5-HTP accumulation by EMD 49,980 is induced mainly via direct activation of 5-HT1A receptors, although some contribution due to 5-HT uptake inhibition is likely. Furthermore, results with various reference compounds make it unlikely that there are indirect effects, also via alpha 2-receptors in the models used and support the view that D-2 agonistic, 5-HT uptake inhibiting and 5-HT1A agonistic actions are independent properties of EMD 49,980. PMID- 2565818 TI - The induction of grooming and vacuous chewing by a series of selective D-1 dopamine receptor agonists: two directions of D-1:D-2 interaction. AB - A range of 3- and 6-substituted 1-phenyl-1H-3-benzazepine analogues of SK&F 38393 with D-1 agonist activity were compared for their behavioural effects in the intact adult rat and for their relative affinities for D-1 and D-2 dopamine receptors in vitro. All compounds showed selective affinity for D-1 receptors and induced prominent grooming behaviour, but those with the lower D-1:D-2 selectivity ratios also induced additional episodes of non-stereotyped sniffing, locomotion and rearing. No vacuous chewing was noted. There were marked differences in in vivo potency, extending over a 100-fold range. These responses to the most potent agonist, SK&F 77434 (3N-allyl-SK&F 38393) were reduced enantioselectively by the D-1 antagonist R-SK&F 83566. They were also reduced enantioselectively by the D-2 antagonist R-piquindone, but this pretreatment additionally released a marked vacuous chewing response to SK&F 77434. Prominent grooming may be a characteristic behavioural response to a range of D-1 agonists. It is suggested that there may be at least two forms of functional interaction between D-1 and D-2 systems, manifested concurrently in distinct elements of behaviour: one co-operative, as in the regulation of grooming, and with correlates in the regulation of pallidal neural activity; the other oppositional, as in the regulation of vacuous chewing, and with correlates in the regulation of striatal adenylate cyclase activity. PMID- 2565819 TI - Role of dopamine D-1 and D-2 receptors in the regulation of neurotensin systems of the neostriatum and the nucleus accumbens. AB - Activation of dopamine D-1 receptors with multiple administrations of SKF 38393 significantly increased the level of neurotensin-like immunoreactivity in the striatum and the nucleus accumbens. However, a similar treatment with the D-2 receptor-selective agonist, LY 171555, decreased the same in both structures; when the two drugs were administered concurrently, their individual effects were blocked. These results suggest that dopamine D-1 and D-2 receptors antagonistically regulate neurotensin systems of the striatum and nucleus accumbens. On the other hand, blockade of D-2 receptors (with sulpiride) elevated, while D-1 receptors blockade (with SCH 23390) caused no change in the level of neurotensin in both these structures. Dopamine D-1 receptors did not appear to contribute to the sulpiride-mediated effect as concurrent administration of SCH 23390 did not alter the response. PMID- 2565820 TI - Expression of dipeptidyl peptidase IV in rat tissues is mainly regulated at the mRNA levels. AB - Dipeptidyl peptidase IV (DPPIV) is a serine peptidase that cleaves N-terminal dipeptides from polypeptides when the second residue is a proline or an alanine. We have recently cloned cDNAs for rat gp110, a membrane glycoprotein with Mr of 110,000 isolated initially from rat liver. Studies reported here establish that the gp110 for which we have cloned cDNAs is DPPIV. Using the antibodies against and cDNA for DPPIV, we have assessed the tissue distribution of DPPIV by molecular approaches. Immunoblot analysis demonstrated that DPPIV is present in the kidney, lung, and small intestine at high levels, in the liver and spleen at moderate levels, and in the heart at low levels. The highest levels of mRNA for DPPIV were detected in the kidney and small intestine as compared to moderate levels found in the lung, liver, and spleen. The lowest levels of DPPIV mRNA were found in the stomach, testis, and heart. No detectable DPPIV protein and mRNA were found in brain or muscle. LDPPIV protein and mRNA are present at much lower levels in fetal livers as compared to the adult liver. Indirect immunofluorescence microscopy demonstrated that DPPIV is localized in the bile canaliculus of hematocytes and in the apical membrane domains of kidney tubule and small intestine. Further studies by Southern blot analysis indicate that DPPIV is encoded by a single gene. PMID- 2565822 TI - Clones containing variant forms of complete human rRNA genes. Characterization and sequence of their transcription initiation region. AB - In order to assess the extent of structural heterogeneity among ribosomal genes in the human genome, several cosmid clones, each containing an entire transcription unit, have been isolated and analyzed. Five cloned genes were recognized as structurally different from each other to some extent, either within the transcription unit or in its immediate vicinity. The sequence of a 1.2 kb region encompassing the transcription start site has been determined for the five cloned genes. Point differences among the genes are observed at five nucleotide positions within the analyzed portion of the 5' external transcribed spacer of the ribosomal gene. Moreover, upstream from the transcription start site, a unique difference is observed with the absence from the three genes of a 19-nucleotide long stretch which is present in the two other variant gene forms. PMID- 2565821 TI - Defect in phorbol acetate-induced translocation of diacylglycerol kinase in erbB transformed fibroblast cells. AB - We here show that tetradecanoyl phorbol acetate (TPA) and 1-oleoyl 2-acetyl glycerol (OAG) cause the translocation of diacylglycerol (DG) kinase from the cytosol to the membrane fractions in chick embryo fibroblast (CEF) cells. However, this translocation is not marked in erbB-transformed chick embryo fibroblast (GEV) cells. The activities of phosphatidylinositol (PI) and phosphatidylinositol 4-phosphate (PIP) kinases in membrane fractions are not altered by TPA treatment in either CEF or GEV cells. Such reduced translocation of DG kinase by TPA is also observed in src-transformed cells, but not in myc transformed cells. These results suggest that the defect in DG kinase translocation may result in failure to suppress the overactivation of protein kinase C in erbB-2 and src-transformed cells, which may lead to cell growth and transformation. PMID- 2565823 TI - Glycolysis in P-glycoprotein-overexpressing human tumor cell lines. Effects of resistance-modifying agents. AB - We show that drugs, such as verapamil, which reverse multidrug resistance (MDR), in P-glycoprotein-overexpressing tumor cells, increased the rate of lactate production in four human MDR cell lines, but not in the parent, sensitive cell lines. The effect on glycolytic rate was maximal at a medium concentration of 2 microM verapamil. The glycolytic rate in sensitive (A2780) and MDR 2780AD) cells showed the same pH dependence, but the effect of verapamil was seen only in 2780AD cells at all pH values investigated (6.6, 7.4 and 8.2). A series of drugs such as nigericin, oligomycin, amiloride and monensin had similar effects in the two cells. Phorbol myristate acetate increased lactate formation in neither cell line. Verapamil induced an extra amount of ATP consumption in P-glycoprotein expressing 2780AD cells of approx. 25 pmol/s per 10(6) cells, which was estimated to be about 10% of cellular energy turnover. PMID- 2565825 TI - Developmental characteristics of fetal arrhythmias during the period from intrauterine to early extrauterine life assessed using dual echocardiography. AB - To evaluate the outcome of fetal arrhythmias, serial echocardiographic examinations were made on 29 fetuses between 18 and 41 weeks gestation, including 23 with premature contractions (PCs) and 6 with complete atrioventricular block (AVB). Of the cases of PCs 43.5% (10/23) disappeared spontaneously during the antenatal period. 39.1% (9/23) converted to a normal sinus rhythm in early neonatal life, while 17.4% (4/23) had complications of either atrial or ventricular tachycardia. All with AVB (6/6) remained, regardless of advance in gestational age and continued through to the postnatal period. PCs and AVB indicate functional error and an impairment during development of the conduction system, respectively. PMID- 2565824 TI - [Studies on the detection of antibodies to Neisseria gonorrheae using pilus antigen]. AB - The formation of antibodies against pilus antigen of strain F-62 was studied by indirect hemagglutination in the sera of 100 patients. The criterion for the diagnosis of gonorrheal disease was a positive culture of Neisseria gonorrhoeae. A positive smear test (staining with methylene blue) was accepted only in a few cases if the chain of infection could be followed completely. Retrospective studies showed that 75% of our patients with gonorrheal disease had antibodies against pilus antigen. The specificity was 91.7%. Attention is drawn to problems involved in evaluating the test results. PMID- 2565826 TI - Torsion of ovarian tumors: a clinicopathological study. AB - Torsion of ovarian tumors occurred predominantly in the reproductive age group. The majority of the cases presented in pregnant (22.7%) than in non-pregnant (6.1%) women. The major presenting symptom was pain but an abdominal mass was palpable in 79.4% of cases. Torsion was more common on the right ovary and 50% were gangrenous at laparotomy. Most of the tumors were benign cystic teratomas. Only 8.7% of the tumors were malignant. PMID- 2565827 TI - Two-stage repair of giant vesico-vaginal fistula. AB - A review of 840 bladder fistulae over 10 years revealed that 64 (7.6%) were giant vesico-vaginal fistulae. Association with vaginal stenosis and recto-vaginal fistulae were 23.4% and 4.6%, respectively. The results of direct one-stage repair were poor at 35% for the first attempt while the two-stage repair achieved 80% success at the first attempt. Advantages of the two-stage repair include change in the direction of repair for maximum protection of the terminal ureters and also the possibility of change in patient positioning and approach to the repair of the fistula after the first stage. PMID- 2565828 TI - Dating sonographic endometrial images in the normal ovulatory cycle. AB - Pelvic sonograms were obtained from 5 fertile volunteers and 150 infertile patients with normal ovulatory cycles, using real-time ultrasound. The existence of a linear cavity echo (C), thick hypoechoic layer (H) and increased echogenicity (E) were chosen as the parameters of sonographic endometrial images to be studied for dating the ovulatory cycle. In 109 serial sonograms from the fertile volunteers, H was observed from 3 days before ovulation (day -3) to ovulation day (day 0). E was observed from day +1 to day +8, and C, from day -10 to day +3. Studies on 189 sonograms from the infertile patients revealed a similar pattern for these parameters. We tested the accuracy of sonographic images of the endometrium, using sonograms from five patients who underwent hysterectomy. The measurements of endometrial thickness, in vivo, and in vitro, showed little difference. Sonographic endometrial images are considered indicative of histological changes, under the influence of estradiol and progesterone. Therefore, observation of the combined quantitative changes in C, H and E facilitates dating of the sonographic endometrial images in a normal ovulatory cycle. PMID- 2565829 TI - Effect of immunopotentialization on rate of vaginal smear normalization according to appearance of cervical intraepithelial neoplasia. AB - Sixty one women (age range 20-35 years) treated by Gynatren immunopotentialization (three vaccinations every second week) were divided into two groups: group I with cervical intraepithelial neoplasia (CIN) and group II without CIN. Four weeks of therapy resulted in normalization of the vaginal flora and cervical milieu as well as in disappearance of CIN in 54% of cases stated cytologically. These findings confirmed our previous observations. Moreover, it was stated that process of regression of inflammation, metaplasia as well as normalization of vaginal flora is less apparent in women with CIN. PMID- 2565830 TI - Progesterone in saliva as an index of ovarian function. AB - Progesterone was assayed in samples of saliva collected daily throughout the menstrual cycle and correlated with corresponding daily concentrations of pregnanediol-3 alpha-glucuronide in early morning urine. Ovarian follicular growth was monitored on a daily basis with a real-time sector scanner. The concentrations of salivary progesterone shows a specific pattern with a mid-cycle rise and a peak in the luteal phase. The mean concentration in the follicular phase, as determined, was 40.4 pmol/l (range 15.3-110.7 pmol/l). It increased in the peri-ovulatory period to a peak of 201.1 pmol/l (range 46.4-289.8 pmol/l) 6 days following follicular rupture. PMID- 2565831 TI - Labor outcome of juvenile primiparae in a population with a high incidence of contracted pelvis. AB - Labor outcome of primiparae less than 17 years was compared with non-juvenile primiparae in a population with a high incidence of contracted pelvis. Juvenile primiparae were referred to hospital on the basis of age, whereas non-juveniles were referred for an obstetric complication. There was no statistical difference in rates of cesarean section, Ventouse, operative delivery, low birth weight, or perinatal mortality between 538 juveniles and 5294 older "high risk" mothers. Symphysiotomy rates were actually higher in the juveniles (12.6% vs. 9.7%, P less than 0.05). These data support the practice of hospital referral for juvenile African women in labor. PMID- 2565833 TI - Malignant ovarian tumors associated with pregnancy: report of six cases. AB - Six cases of pregnancy associated malignant ovarian tumors, four epithelial cancers, one immature teratoma and one metastatic cancer of colon origin, are reported. One patient with mucinous cystadenocarcinoma had a history of persistent ovarian tumor during her past three pregnancies. Another patient was found to have mucinous cystadenocarcinoma after an emergency operation for twisted ovarian tumor. Immature teratoma associated with pregnancy is very rare and our case seems to be the 8th reported such case. PMID- 2565832 TI - Radionuclide hysterosalpingography for measurement of human oviductal function. AB - A clinical method is described and claimed to evaluate the active transportation capacity of the luminal epithelium lining the human uterus and fallopian tube. After intracervical application of technetium-labelled human albumin spheres at 1 2 days before ovulation this radioactivity could be followed by gamma-camera when moved in ad ovarian direction. This method is called radionuclide hysterosalpingography (RN-HSG). The results were compared to the findings at normal hysterosalpingography (HSG). By use of this method it was possible to verify active passage in cases of tubal spasm at HSG, lack of transport in cases of normal patent oviducts at HSG as well as presence or absence of active transport through sactosalpinges with or without fimbrial passages to the abdominal cavity as seen at normal HSG. Congruent findings between HSG and RN-HSG was observed in 49%. The studied oviducts were found to be patent with normal HSG but lacked transportation capacity when studied by RN-HSG in 41%. The clinical use of this method is discussed in view of the selection of patients for different forms of in vitro fertilization and egg transfer (IVF-ET). PMID- 2565834 TI - Endometrial carcinoma presenting as a large inguinal mass. AB - Patients with endometrial cancer usually present with vaginal bleeding. A case of endometrial cancer presenting as an asymptomatic, large, inguinal mass is reported. This unusual clinical picture and its management are discussed. PMID- 2565835 TI - Uterine sarcoma with liposarcomatous differentiation: report of a case and review of the literature. AB - Heterologous sarcomas of the uterus are uncommon neoplasms. A liposarcoma in combination with a leiomyosarcoma of the uterus has been reported only once prior to this case report. We report a case of uterine sarcoma demonstrating two distinct variants of liposarcoma. This has not been seen to date. PMID- 2565836 TI - Postcoital vesico-vaginal fistula following surgery for cancer cervix. AB - A 25-year-old parous woman was treated with radical hysterectomy for cancer of the cervix. Six months later she developed a vesico-vaginal fistula following coitus. The fistula was repaired and vaginal reconstruction performed. Sexual dysfunction and the need for vaginal reconstruction are discussed. PMID- 2565837 TI - Urinary excretion of N-acetyl-beta-D-glucosaminidase and alanine aminopeptidase during pregnancy. AB - The assay of enzyme activity in urine seems a reliable and safe method to monitor different kidney diseases. However, its use in pregnant patients might be limited by the modifications of kidney function during pregnancy. The aim of the present study was to evaluate the trend of excretion of the lysosomal enzyme N-acetyl beta-D-glucosaminidase (NAG) and the brush border enzyme alanine aminopeptidase (AAP) during uncomplicated pregnancies. NAG excretion showed a significant increase (P less than 0.001) throughout pregnancy, while no significant modification of AAP levels was demonstrated. These data support the hypothesis that the two enzymes are excreted into the urine through different mechanisms and might constitute markers for different pathological events. As the increase of NAG excretion may be related to the kidney functional adaptation to pregnancy, different cut-off limits must be established in this period. PMID- 2565838 TI - Linkage studies on NIDDM and the insulin and insulin-receptor genes. AB - Twenty Black families in which at least two siblings had non-insulin-dependent diabetes mellitus (NIDDM) were typed for restriction-fragment-length polymorphisms at the insulin (INS), insulin-receptor (INSR), and HLA-DR beta loci. Evidence for linkage between NIDDM and these loci was assessed with various genetic models for the transmission of NIDDM and with the affected-sib-pair approach, which does not require assumptions concerning a genetic model for NIDDM. Tight linkage between NIDDM and any of the loci was unlikely under all of the genetic models examined. Similarly, for all three of the loci, the distribution of affected sib pairs sharing 2, 1, or 0 genes identical by descent was not significantly different from (and was very similar to) that expected if the locus were unrelated to disease susceptibility. There was no evidence for linkage heterogeneity for any of the loci when families were grouped according to obesity or age at onset or when considering families individually. We conclude that the INS and INSR loci can be ruled out as major susceptibility loci for NIDDM in most Black families segregating this disorder, but we recognize that defects at either of these loci may cause or contribute to NIDDM in some patients. In addition, it is possible that variation at the INS and/or INSR loci may contribute to NIDDM susceptibility by modifying susceptibility due primarily to another major gene(s) or as part of an overall polygenic component to NIDDM. PMID- 2565839 TI - Pharmacological basis for the use of nimodipine in central nervous system disorders. AB - Nimodipine, a Ca2+ antagonist with cerebrovasodilatory and anti-ischemic effects, binds to rat, guinea pig, and human brain membranes with high affinity (less than 1 nM). Only at higher concentrations has nimodipine been reported to block the release of some neurotransmitters and hormones from neuronal tissue. Nimodipine has no consistent effect on brain oxygen consumption or cortical ATP or phosphocreatine levels, although the ischemia-induced fall of brain ATP levels in gerbils or the lowering of intracellular brain pH in rabbits with focal cerebral ischemia were antagonized by the drug. In rats and baboons with middle cerebral artery occlusion, nimodipine was found to reduce neurological deficits without an increase in intracranial pressure or brain edema. Electrophysiological studies with nimodipine suggested a direct neuronal action. In rabbit dorsal root ganglion cells, concentrations as low as 20 nM were reported to block inward Ca2+ currents. Recent studies have suggested that nimodipine may also improve memory in brain-damaged or old rats, restore sensorimotor function and abnormal walking patterns of old rats, and accelerate acquisition of associative learning in aging rabbits. Blockade of age-related changes in Ca2+ fluxes in rat hippocampal neurones by nimodipine in vitro pointed to neuronal plasma membrane as the site of nimodipine action. The therapeutic usefulness of nimodipine appears not to be limited to cerebral ischemia, but may include dementia, age-related degenerative diseases, epilepsy, and ethanol intoxication. PMID- 2565840 TI - Brevetoxins: unique polyether dinoflagellate toxins. AB - Brevetoxins are lipid-soluble polyether marine toxins of unique structure and pharmacological function. Toxins are active in vivo in the nanomolar to picomolar concentration range and in vitro in isolated neuromuscular or giant axon preparations and in single-cell or subcellular model systems. Their effect is excitatory, mediated by the enhancement of cellular Na+ influx. Brevetoxins bind at site 5 on the voltage-sensitive sodium channel, a specificity shared with ciguatoxin. This site is allosterically linked to other natural toxin binding sites on the channel. PMID- 2565841 TI - Negative effects of famotidine on cardiac performance assessed by noninvasive hemodynamic measurements. AB - In a randomized placebo-controlled study 12 healthy volunteers were treated for 1 wk each with 10 mg of nifedipine four times daily plus placebo or the same dose of nifedipine concurrently with 40 mg of famotidine once a day. Famotidine did not significantly alter pharmacokinetic parameters of nifedipine. Determination of systolic time intervals showed that the preejection period and the ratio of the preejection period and the left ventricular ejection time were significantly reduced by administration of nifedipine plus placebo. Coadministration of famotidine and nifedipine, however, led to a significant increase of these parameters. In an additional double-blind study, a significant rise of the preejection period and of the ratio was detected after administration of famotidine alone. In impedance cardiography stroke volume and cardiac output were significantly reduced by famotidine. Heart rate and blood pressure values were not altered by the H2-antagonist. For the first time, to our knowledge, the observed changes of hemodynamic parameters appear to indicate that famotidine may exert negative effects on cardiac performance which, in our opinion, could be of clinical relevance in elderly subjects or in patients with heart failure. PMID- 2565842 TI - Parenteral antisecretory drug therapy in patients with Zollinger-Ellison syndrome. AB - Forty-six patients with Zollinger-Ellison syndrome were studied prospectively to determine a safe and effective method and criterion for controlling gastric acid hypersecretion during periods when oral antisecretory agents could not be used. In each patient it was possible to reduce acid secretion to less than or equal to 10 mEq/h after an i.v. bolus of 150 or 300 mg of cimetidine and a stepwise titration of cimetidine given by continuous infusion. The mean dose given by i.v. infusion was 2.9 mg/kg body wt.h but there was a wide range (0.5-7.0 mg/kg body wt.h) and the minimal dose had to be determined individually for each patient. The minimal i.v. cimetidine dose did not correlate with basal or maximal acid output or fasting gastrin concentration, but correlated closely with either the previous oral dose of cimetidine (r = 0.96, p less than 0.001) or the previous oral dose of ranitidine or famotidine (r = 0.95, p less than 0.001). To study the efficacy and safety of an i.v. infusion of cimetidine, 34 patients undergoing surgery were maintained on i.v. cimetidine for a mean of 12 days (range 1-83 days). One-half of the patients did not require dose adjustment, whereas the remainder required an average of 2 adjustments, usually in the first 3 postoperative days. No patient developed complications attributable to gastric acid hypersecretion in the postoperative period, and there was no detectable neurologic, hematologic, or hepatic toxicity. This study demonstrates that a continuous i.v. infusion of cimetidine adequately inhibits gastric acid hypersecretion in patients with Zollinger-Ellison syndrome. However, high doses were frequently required, the dose had to be determined in a stepwise fashion individually for each patient, and the i.v. dose correlated with the previous oral dose. Reducing acid secretion to less than or equal to 10 mEq/h was a safe criterion during surgery and continuous i.v. cimetidine was safe and effective in achieving this degree of control for up to 83 days. PMID- 2565844 TI - Costs of medical and surgical treatment of duodenal ulcer. AB - Proximal gastric vagotomy and intermittent and maintenance therapy with H2 antagonists have all been claimed to be effective in long-term management of duodenal ulcer disease. The model of a Markov chain was used to compare their costs by a medical decision analysis. The high price of the initial procedure made proximal gastric vagotomy the most expensive therapy, its costs rising from +10,600 after 1 yr to +12,200 after 15 yr. The average costs of intermittent therapy per patient rose from +500 to +7500. Maintenance therapy cost as much as intermittent therapy but provided 8% and 4% more time spent free of ulcer relapse and pain, respectively. In a sensitivity analysis, the order of the therapeutic options regarding their cost-effectiveness remained robust to changes in the assumptions underlying the model. In a European health care system, the initial surgical procedure cost only one-seventh of the average annual income compared with two-thirds in the United States, and proximal gastric vagotomy turned out to be the cheapest therapy after 6 yr. These results suggest that maintenance therapy provides the best long-term management. Gastric surgery may represent a cost-effective measure of ulcer prevention in Europe but not in the United States. PMID- 2565843 TI - Mechanism for increase of gastrin release by secretin in Zollinger-Ellison syndrome. AB - In patients with Zollinger-Ellison syndrome, serum gastrin level is increased by secretin and is decreased by somatostatin. To elucidate the cellular mechanism for these actions, we investigated the direct effects of secretin and somatostatin on dispersed gastrinoma cells from a patient with Zollinger-Ellison syndrome. In the presence of 3-isobutyl-1-methylxanthine, secretin significantly stimulated gastrin release from dispersed gastrinoma cells, which was inhibited by somatostatin. In the presence of guanosine 5'-triphosphate, furthermore, secretin enhanced adenylate cyclase activation in the membranes from these cells, and this activation was reduced by somatostatin, whereas neither secretin nor somatostatin affected inositol phospholipid turnover. On the other hand, removal of guanosine 5'-triphosphate from incubation medium abolished both the stimulatory effect of secretin and the inhibitory effect of somatostatin on adenylate cyclase activation. Furthermore, pertussis toxin pretreatment reversed the ability of somatostatin to inhibit secretin-induced increase in gastrin release and activation of adenylate cyclase. Thus, in this gastrinoma patient, secretin and somatostatin appeared to act directly on gastrinoma cells to stimulate and inhibit gastrin secretion, respectively, by modulating adenylate cyclase activation, probably via guanine nucleotide-binding proteins. PMID- 2565845 TI - Premedication for upper gastrointestinal endoscopy. AB - Premedication is not essential to endoscopy but patient tolerance is clearly improved and, thus, ease of examination. Although comparable results can be achieved through nonpharmacologic means, the time and effort involved precludes their widespread use. Despite near universal utilization of premedication in endoscopy, the associated risk is difficult to determine from the available literature. The reported data reveal nominal risk yet must be viewed as minimums. The ideal drug with predictable clinical effects, minimal postprocedure impairment, little respiratory compromise, and proven antagonist is not yet available. Although midazolam seems to represent an advance, recent emphasis on respiratory depression is particularly troublesome. Studies evaluating various agents have suffered from lack of quantitation of such parameters as patient tolerance, ease of examination, and postprocedure impairment. Development of proven standards for these parameters would have to occur before a definitive double-blind randomized trial could be undertaken. Suggested means of assessing these parameters are listed in Table I. Improvement in major morbidity would be difficult in light of its low incidence. As the search for the ideal drug continues, endoscopists must continue to use drugs whose full effects are incompletely understood. The ability to increase patient comfort must be balanced with the small, but ever present, risk of morbidity and mortality. PMID- 2565846 TI - Anti-acid secretion activity of drugs cimetidine, ranitidine, tiotidine D 15,144 in dogs fixed with gastric fistulae. AB - 1. Acid secretion for each dog has reached a near maximum (100%) at the 6th samples, 90 min after the intravenous infusion of histamine (10 mu ghr-1, or approximately equal to 0.3 mghr-1). 2. 0.5 mgkg-1 Cimetidine had produced a mean inhibition of 47% on the stomach. 3. 0.1 mgkg-1 Ranitidine (D 14,951) could only inhibit a maximum of 28%, and the secretion had return to normal in just 30 min. 4. 0.025 mgkg-1 Tiotidine (D 15,104) had inhibited 53% acid secretion within 15 min of exposure. Recovery was quite similar to that of Cimetidine, at 150 min. 5. At a dosage one fifth of Cimetidine (0.1 mgkg-1) D 15,144 had depressed 35% of acid secretion at the first 15 min. The inhibition is gradually increased to about 43% (at 30 min), and was maintained for the next 105 min. PMID- 2565847 TI - Short time action of antirheumatic substances on the liver of rat. Protective effect of tryptophan + methionine. AB - 1. A large number of drugs, including some antirheumatic substances, cause liver damage. 2. Only a little information is available, so far, on the causes of such damage and their prevention. 3. From studies on a number of antirheumatic drugs as to their effect upon the liver, three categories could be differentiated: (a) large effect (b) low effect (c) no effect. 4. Judging from our results, the liver damage is induced via a disturbance of the NAD-adenoribosylation metabolism. PMID- 2565848 TI - Effect of histamine on two different affinity sites in beta-adrenoceptors. AB - 1. To study the effect of histamine on two different affinity sites in beta adrenoceptors, we tested specific binding of [3H]befunolol to microsomal fractions from the guinea pig taenia caecum in the presence and absence of histamine (10(-5) M). 2. The Scatchard plot of data in the absence of histamine was recognized as two straight lines suggesting the existence of two different affinity sites: high and low. However, a curvilinear plot with upward concavity was observed in the presence of histamine (10(-5) M). 3. These results suggest the possibility that histamine alters the binding sites of beta-adrenoceptors into several classes of sites with lower affinity. 4. Further, an antagonism between isoprenaline and BFE-55, which interacts only with the high affinity site, was tested on the atria (beta 1-adrenoceptor predominant) and tracheae (beta 2-adrenoceptor predominant) of the guinea pig. The pA2-values in the atria were almost equal to those in the trachea, suggesting that both beta 1- and beta 2-adrenoceptors contained two different affinity sites. PMID- 2565849 TI - Effects of TRH and DN-1417 on high potassium-evoked acetylcholine release from rat basal forebrain slices determined directly by radioimmunoassay. AB - 1. High potassium (50 mM)-evoked acetylcholine (ACh) release from rat basal forebrain slices under conditions without an exogenous choline supply was determined using a radioimmunoassay for ACh. 2. A consistent amount of ACh release was observed at each repetitive stimulation and ACh content in brain slices was not altered by potassium stimulations. These results indicate the existence of a large intracellular releasable ACh store, which is independent of new synthesis from exogenous choline. 3. Atropine, even at a concentration of 10( 6) M, did not affect the potassium-evoked ACh release. Thus, modulation of ACh release by the muscarinic autoreceptor was not revealed under the conditions employed. 4. Thyrotropin-releasing hormone (TRH, 10(-4) M) caused a slight and statistically insignificant increase in potassium-evoked ACh release. DN-1417, a TRH analogue, at a concentration of 10(-4) M significantly increased potassium evoked ACh release. These findings indicate that DN-1417 is able to enhance ACh output independently of ACh synthesis from exogenous choline. PMID- 2565850 TI - Kappa-receptor mechanisms in synaptosomal Ca uptake. AB - 1. Inhibition of Ca uptake by certain opioids was tested in synaptosomes of rat brain. The potency order was dynorphin A 1-13, a kappa-selective agonist greater than nalorphine greater than nalorphine epoxide greater than morphine. 2. The pA2 values (negative logarithms of dissociation constant) of naloxone against four opioids were not significantly different from each other, suggesting that the site of action of the four opioids is identical. 3. Morphiceptin, a mu-selective agonist and DADLE, a delta-selective agonist had no effect on Ca uptake. 4. These results suggest that the site of action of the four opioids is kappa-receptors. 5. Potency order estimated from competition inhibition curves of specific binding of [3H]ethylketo-cyclazocine (kappa-selective ligand) by the test opioids was nalorphine greater than nalorphine epoxide greater than dynorphin A 1-13 greater than morphine. 6. The difference between the two potency orders suggests that affinities and intrinsic activities of the drugs are important factors in determining their agonistic activity in kappa-receptor mechanisms. PMID- 2565851 TI - Growth hormone and somatostatin in the plasma of transiently diabetic ducks: basal variation and response to glucose. AB - In the duck, subtotal pancreatectomy induces a transient diabetes, with decreased insulin and glucagon basal levels as well as responses to glucose. At the same time, a transient increase in basal peripheral somatostatin occurs, followed by an increase in growth hormone in the postdiabetic state. Intravenous glucose induces a slight decrease in somatostatin secretion in normal, but not in diabetic animals, and no significant variation in growth hormone secretion at any state. An obvious role of growth hormone or somatostatin in the development of this transient diabetes in the duck could not be detected in this study. PMID- 2565852 TI - A novel fimbrial haemagglutinin produced by a strain of Salmonella of serotype Salinatis. AB - A strain of Salmonella of serotype Salinatis, that produced a mannose-resistant and eluting haemagglutinin (MREHA) when cultured at 37 degrees C but not at 18 degrees C, was examined by electron microscopy after negative staining. Production of this MREHA, previously described as being non-fimbrial, was correlated with the presence of thin fimbriae which had an external diameter of 3.6 nm. The purified Salinatis thin fimbriae had an estimated Mr of 19 kDa. This fimbrial MREHA was not produced by strains of the antigenically related serotypes Duisburg and Sandiego. PMID- 2565853 TI - The complete nucleotide sequence of the glutamine synthetase gene (glnA) of Bacillus subtilis. AB - The glutamine synthetase (GS) gene from Bacillus subtilis PCI 219 was cloned in Escherichia coli using the vector pBR329. A plasmid, pSGS2, was isolated from a glnA+ transformant and the cloned GS gene was found to be located in a 3.6 kb DNA fragment. The nucleotide sequence of a 1.8 kb segment encoding the GS was determined. This segment showed an open reading frame which would encode a polypeptide of 444 amino acids. The amino acid sequence of this GS gene product has higher homology with that of the Clostridium acetobutylicum GS than that of the E. coli GS. PMID- 2565854 TI - Molecular genetics of the Caenorhabditis elegans heterochronic gene lin-14. AB - We describe a general strategy for the genetic mapping in parallel of multiple restriction fragment length polymorphism (RFLP) loci. This approach allows the systematic identification for cloning of physical genetic loci within about 100 kb of any gene in Caenorhabditis elegans. We have used this strategy of parallel RFLP mapping to clone the heterochronic gene lin-14, which controls the timing and sequence of many C. elegans postembryonic developmental events. We found that of about 400 polymorphic loci in the C. elegans genome associated with the Tc1 family of repetitive elements, six are within 0.3 map unit of lin-14. The three closest lin-14-linked Tc1-containing restriction fragments were cloned and used to identify by hybridization an 830-kb region of contiguous cloned DNA fragments assembled from cosmid and yeast artificial chromosome libraries. A lin-14 intragenic recombinant that separated a previously cryptic lin-14 semidominant mutation from a cis-acting lin-14 suppressor mutation was used to map the location of the lin-14 gene to a 25-kb region of this 830-kb contig. DNA probes from this region detected lin-14 allele-specific DNA alterations and a lin-14 mRNA. Two lin-14 semi-dominant alleles, which cause temporally inappropriate lin 14 gene activity and lead to the reiterated expression of specific early developmental events, were shown to delete sequences from the lin-14 gene and mRNA. These deletions may define cis-acting sequences responsible for the temporal regulation of lin-14. PMID- 2565856 TI - Mapping RFLP loci in maize using B-A translocations. AB - Plants hypoploid for specific segments of each of the maize (Zea mays L.) chromosomes were generated using 24 different B-A translocations. Plants carrying each of the B-A translocations were crossed as male parents to inbreds, and sibling progeny hypoploid or not hypoploid for specific chromosomal segments were recovered. Genomic DNAs from the parents, hypoploid progeny, and nonhypoploid (euploid or hyperploid) progeny for each of these B-A translocations were digested with restriction enzymes, electrophoresed in agarose gels, blotted onto reusable nylon membranes, and probed with nick-translated, cloned DNA fragments which had been mapped previously by restriction fragment length polymorphism (RFLP) analysis to the chromosome involved in the B-A translocation. The chromosomal segment on our RFLP map which was uncovered by each of the B-A translocations was determined. This work unequivocally identified the short and long arms of each chromosome on this map, and it also identified the region on each chromosome which contains the centromere. Because the breakpoints of the B-a translocations were previously known on the cytological and the conventional genetic maps, this study also allowed this RFLP map to be more highly correlated with these maps. PMID- 2565855 TI - Molecular population genetics of mtDNA size variation in crickets. AB - Nucleotide sequence analysis of a region of cricket (Gryllus firmus) mtDNA showing discrete length variation revealed tandemly repeated sequences 220 base pairs (bp) in length. The repeats consist of 206 bp sequences bounded by the dyad symmetric sequence 5'GGGGGCATGCCCCC3'. The sequence data showed that mtDNA size variation in this species is due to variation in the number of copies of tandem repeats. Southern blot analysis was used to document the frequency of crickets heteroplasmic for two or more different-sized mtDNAs. In New England populations of G. firmus and a close relative Gryllus pennsylvanicus approximately 60% of the former and 45% of the latter were heteroplasmic. From densitometry of autoradiographs the frequencies of mtDNA size classes were determined for the population samples and are shown to very different in the two species. However, in populations where hybridization between the two species has occurred, the frequencies of size classes and cytoplasmic genotypes in each species' distinct mtDNA lineage were shifted in a manner suggesting nuclear-cytoplasmic interactions. The data were applied to reported diversity indices and hierarchical statistics. The hierarchical statistics indicated that the greatest proportion of variation for mtDNA size was due to variation among individuals in their cytoplasmic genotypes (heteroplasmic or homoplasmic state). The diversity indices were used to estimate a per-generation mutation rate for size variants of 10(-4). The data are discussed in light of the relationship between genetic drift and mutation in maintaining variation for mtDNA size. PMID- 2565857 TI - The Hox-1.3 homeo box protein is a sequence-specific DNA-binding phosphoprotein. AB - We report that the murine Hox-1.3 homeo domain protein is a nuclear phosphoprotein capable of binding to specific DNA sequences. DNase I protection of the Hox-1.3 gene promoter region with the Hox-1.3 protein identifies a binding site 144 bp upstream from the start of transcription. Both phosphorylated and nonphosphorylated forms bind DNA directly in a sequence-specific manner. Electrophoretic mobility shift assays were performed with a set of synthetic oligonucleotides representing either the DNase I-protected region of the Hox-1.3 gene or partially homologous sequences present in promoter regions of other characterized viral, yeast, and mammalian genes. From the results, we deduce a consensus binding motif of CPyPyNATTAT/GPy. Base substitutions in the core ATTA sequence severely reduce or abolish binding. In the SV40 enhancer, the Hox-1.3 binding motif overlaps both the octamer (Octa2) and the transactivator protein-1 (AP-1) binding sites. The Hox-1.3 binding motif also overlaps the nuclear factor III (NF-III) octamer motif in the adenovirus-2 origin of DNA replication. Overlap among DNA-binding sites suggests that regulation imparted by certain cis-elements may be integrated by these different factors. PMID- 2565858 TI - Structure and expression of a family of Ultrabithorax mRNAs generated by alternative splicing and polyadenylation in Drosophila. AB - The 77-kb primary transcript of the homeotic Ultrabithorax (Ubx) gene is alternatively spliced to yield at least five different coding regions. Each is restricted to either a 3.2- or a 4.3-kb size class generated by alternative polyadenylation. The pathways for splicing and polyadenylation are therefore coordinately regulated, and because the relative abundance of the respective mRNAs varies throughout development, these pathways also appear to be developmentally regulated. Translation of these mRNAs yields a family of Ubx proteins characterized by constant amino- and carboxy-proximal regions of 247 and 99 amino acid residues, respectively. Members of this family are distinguished by a short variable region that links the constant regions and consists of different combinations of three optional elements of 9, 17, and 17 residues. Only four amino acid residues separate this variable region from the 60-residue homeo domain of the carboxy-terminal constant region. This proximity suggests that functional differences among the Ubx proteins derive from the differential effects of their variable regions on the DNA-binding capacity of the homeo domain. An argument is made that these functional differences are tissue specific. PMID- 2565859 TI - [Various subjective and objective indicators of intellectual and emotional stress in air traffic controllers during different work shifts]. AB - The purpose of the study was to examine air-traffic controllers attending the automated air-traffic management system called Start. The data revealed that during the night shift air-traffic controllers experienced significant changes in body functioning in comparison with morning and day shifts (deterioration of self appraisal indices and elevation of catecholamin excretion). The correlation analysis demonstrated that activation of sympathoardrenal chains (primarily of the mediator chain) had a negative influence on the indices of health state, activity and attitude of mind of air-traffic controllers. PMID- 2565860 TI - Rebound nocturnal hypersecretion after four weeks treatment with an H2 receptor antagonist. AB - Daytime intragastric pH, fasting and meal stimulated serum gastrin and nocturnal acid output were studied in eight male duodenal ulcer patients before, during and two days after completing nizatidine 300 mg nocte (20:00 h) for four weeks. Median nocturnal acid output (mmol/10 h) decreased during treatment to 11.6 (range 0.4-26.7) compared with pretreatment value of 39.4 (9.8-91.2); median acid inhibition 77% (p less than 0.01) which was strongest between 24:00 and 04:00 h. Two days after discontinuing treatment, nocturnal acid output increased to 74.1 (11-181). Compared with the pretreatment value this represents median rebound hypersecretion of 77% (p less than 0.05), caused by increased H+ concentration and volume of secretion. Overall median daytime intragastric pH (09:00-21:00 h) was unchanged on the final day of treatment and two days after completing therapy, compared with the pretreatment values. Fasting serum gastrin measured between 09:30 and 10:00 h and the integrated gastrin response to an OXO breakfast taken out at 10:00 h were also similar during and after treatment, compared with pretreatment values. The rebound nocturnal hypersecretion may be relevant to the high ulcer relapse rates after stopping H2 receptor antagonists. PMID- 2565861 TI - [Antihypertensive effect of bopindolol on stroke-prone spontaneously hypertensive rats (SHRSP)]. AB - The antihypertensive effect of bopindolol, a long-acting beta-adrenoceptor blocking agent, was investigated in stroke-prone spontaneously hypertensive rats (SHRSP). One group received tap water during the period of 8 to 32 weeks of age. The average dose of bopindolol administered was calculated from water intake to be approximately 1.4 mg/kg/day. The lowering effect in blood pressure of bopindolol was apparent at the age of 14 weeks, and this continued up to the end of the experiment. Bopindolol significantly reduced the heart rate. Plasma levels of urea nitrogen (BUN), triglyceride, and phospholipid of SHRSP treated with bopindolol were lower than those of the control SHRSP. One of the 8 control SHRSP died, and no rats treated with bopindolol died during the experiment. The histopathological study revealed that three of the control SHRSP had cerebral apoplexy, whereas there was no evidence of cerebral apoplexy in the treated SHRSP. Chronic treatment of bopindolol clearly alleviated myocardial fibrosis and hypertrophic changes in the left ventricular wall of the heart. Decreases in the incidence of proliferative arteritis and malignant nephrosclerosis in the kidney and necrotizing arteritis of the mesenteric arteries were observed in SHRSP treated with bopindolol. The data presented indicate that bopindolol is a powerful antihypertensive agent. PMID- 2565862 TI - Bopindolol: long-term effects on blood pressure, renal function, plasma renin activity, and plasma aldosterone in mild-to-moderate essential hypertension. AB - Bopindolol was given to hypertensive patients in a single daily dose ranging from 1 to 4 mg. In 86.7% of the patients diastolic blood pressure was reduced by 10 mmHg or more, or lowered to below 90 mmHg, already during the first 4 wk of treatment. Inulin clearance decreased by 10%; PAH and creatinine clearance, blood urea nitrogen, and serum creatinine were unchanged. PMID- 2565863 TI - Localization of adrenergic and neuropeptide tyrosine-containing nerves in the mammalian liver. AB - The distribution of adrenergic nerves in guinea pig and rat liver was studied by the immunolocalization of fibers containing tyrosine hydroxylase and dopamine beta-hydroxylase, enzymes involved in the synthesis of catecholamines. In both species, adrenergic fibers were identified within portal tracts, often in close proximity to hepatic artery branches. In guinea pig liver, but not rat liver, abundant intraacinar fibers were identified; fibers were also seen within the walls of terminal hepatic vein radicles and larger hepatic veins. The presence of peptidergic nerves containing the regulatory peptide neuropeptide tyrosine and the C-flanking peptide CPON was investigated by indirect immunofluorescence. The distribution of these nerves was similar to that of tyrosine hydroxylase- and dopamine beta-hydroxylase-positive nerves and showed the same species difference. The effector sympathetic nature of tyrosine hydroxylase- and neuropeptide tyrosine-positive fibers in rat liver was confirmed by chemical denervation studies using 6-hydroxydopamine. PMID- 2565864 TI - Physical mapping of DXS134 close to the DXS52 locus. AB - The locus DXS134 (cpX67) has been physically linked to the cluster of polymorphic loci DXS52, DXS15, and DXS33. A comparison of physical and genetic distance indicates a high rate of recombination in this region. PMID- 2565865 TI - Mapping of the gene encoding the multifunctional protein carrying out the first three steps of pyrimidine biosynthesis to human chromosome 2. AB - The CAD gene encodes a trifunctional protein that carries the activities of the first three enzymes (carbamyl phosphate synthetase II, aspartate transcarbamylase, and dihydroorotase) of de novo pyrimidine biosynthesis. Genomic fragments of the human CAD gene have been obtained by screening a human genomic library in bacteriophage lambda using a Syrian hamster cDNA clone as a probe. These human genomic clones have been used to assign the CAD gene to human chromosome 2 using in situ hybridization to human metaphase chromosomes and Southern blot hybridization analysis of DNA isolated from a panel of Chinese hamster/human hybrid cells. In situ hybridization analysis has allowed further localization of this gene to the chromosomal region 2p21-p22. PMID- 2565866 TI - Pseudodeficiency of arylsulfatase A: a common genetic polymorphism with possible disease implications. AB - At the locus for arylsulfatase A (ASA) at least four to five alleles exist: besides the normal ASA+ and at least two to three deficiency alleles (ASA-), a pseudodeficiency allele, ASAp, is known. On SDS-PAGE the ASAp enzyme migrates slightly faster than ASA+. Treatment of extracts from cells with ASA+/ASA+, ASAp/ASAp, or ASA+/ASAp genotypes with endoglycosidase F leads to the same deglycosylated subunit pattern. Presumably the degree of glycosylation is lower in ASAp than in ASA+. In a large-scale screening project we determined a gene frequency of 7.3% for ASAp. Thus, the ASA locus is polymorphic. In seven families, ASAp showed a codominant mode of inheritance with ASA+. Homozygosity for ASAp has no obvious clinical consequences. In subjects with the compound genotype ASA-/ASAp, the residual enzyme activity may fall below a critical threshold, so that the substrate can no longer be hydrolyzed sufficiently. Since these compounds are not so rare (estimated frequency 0.073%), this mechanism could be of importance in neuropsychiatric disorders with late onset. PMID- 2565867 TI - DNA restriction fragment length polymorphisms in differential diagnosis of genetic disease: application in neuromuscular diseases. AB - Three families, in which several male individuals suffer from a hereditary neuromuscular disease, were examined by analysis of naturally occurring restriction fragment length polymorphisms (RFLPs) and by screening for deletions. Originally, differential diagnosis included spinal muscular atrophy (two families) and limb girdle syndrome (one family) or Becker muscular dystrophy. Since deletions were not detectable, an X-chromosomal segment, carrying DNA markers for the dystrophin gene and its flanking regions was reconstructed; this demonstrated Becker muscular dystrophy is the most probable primary cause of illness in these families. Furthermore, the possible carriership of female members of these families could be determined accurately. PMID- 2565868 TI - Heterogeneity of mRNA expression in Italian fucosidosis patients. AB - Genomic DNA and mRNA from six unrelated Italian patients affected with fucosidosis were analyzed using two probes (AF3 and AF11B) of partial length cDNA coding for the lysosomal enzyme alpha-L-fucosidase. DNA from patient 2, digested with EcoRI, showed a variant pattern of hybridization caused by the loss of an EcoRI site. The same patient showed a markedly decreased amount of mRNA on Northern blot hybridization. Among the remaining patients, who showed no variation at the DNA level, two apparently lacked mRNA for alpha-L-fucosidase whereas the other three showed a transcript similar in size and amount to that observed in controls. These data confirm the genetic heterogeneity of the molecular defects causing fucosidosis in Italy. PMID- 2565869 TI - Four DNA polymorphisms in the LDL-receptor gene and their use in diagnosis of familial hypercholesterolemia. AB - To examine the potential usefulness of restriction fragment length polymorphisms (RFLPs) for diagnosis of familial hypercholesterolemia (FH), we determined the genotype of FH patients and their relatives for the Apa1I, NcoI, PvuII and StuI RFLP of the LDL-receptor gene in a sample of German patients attending the Lipid Clinic in Munich. There was no significant difference in the relative allele frequency between the group of FH patients and controls for any of the four polymorphisms. Using linkage analysis, we could determine the four-RFLP haplotypes of 39 defective and 90 normal LDL-receptor genes in 38 FH families. In our sample, defective LDL-receptor genes occur on 6 different chromosomes determined by the four RFLPs. This suggests that at least 6 different genetic defects may cause FH in this sample. RFLPs of the LDL-receptor gene cannot be used to detect FH in individuals; however, appropriate diagnosis can be carried out in more than 90% of families using linkage analysis and these RFLPs. PMID- 2565870 TI - A new polymorphic marker very closely linked to DXS52 in the q28 region of the human X chromosome. AB - We have isolated an X chromosome probe, St35.691 (DXS305), which detects two RFLPs with TaqI and PstI, whose combined heterozygosity is about 60%. This probe has been assigned to Xq28 by physical and genetic mapping and is very closely linked to DXS52, DXS15, and the coagulation factor VIII gene (F8C). The best estimate of the recombination fraction for the DXS52-DXS305 interval is 0.014, with a lod score of 50.1. Multipoint analysis places DXS305 on the same side of F8C as DXS52, but complete ordering of the three loci was not possible with our present data. This highly informative marker should be useful in the precise mapping of the many disease genes that have been assigned to the Xq28 band. PMID- 2565871 TI - Eco RI RFLP in the human thyroid peroxidase (TPO) gene on chromosome 2. AB - The authors describe the source and characteristics of a new probe (pTP02/0.7EP). PMID- 2565872 TI - Remarks on the article "Use of Linkage Equilibrium Data in Prenatal Diagnosis of Cystic Fibrosis" by L. Strain et al. PMID- 2565873 TI - Isolation and chromosomal localization of the human En-2 gene. AB - By low stringency hybridization we have isolated from a human cosmid genomic library sequences homologous with a probe from the Drosophila engrailed gene. Partial nucleotide sequence analysis shows a consensus splice acceptor site followed by an open reading frame (ORF) that can encode 104 amino acids; the first 94 amino acids have 71% identity with the Drosophila engrailed protein. The shared region contains a homeo domain and is within the region of engrailed shared with the Drosophila invected gene and the mouse En-1 and En-2 genes. At the amino acid level, the human sequence is 85% identical with the mouse En-1 gene and 100% identical with the mouse En-2 gene. Hybridization against a panel of human-hamster somatic cell hybrids maps this human En-2 gene to chromosome 7, and regional mapping by in situ hybridization to human chromosomes localizes it to region 7q36 at the end of the long arm. PMID- 2565874 TI - Development of multiple endocrine neoplasia type 2A does not involve substantial deletions of chromosome 10. AB - In MEN2A both familial and sporadic cases are known. The familial cases show a dominant pattern of inheritance. In these respects, MEN2A resembles other tumors in whose etiology so-called tumor suppressor genes play a decisive role. The MEN2A locus has been assigned to chromosome 10 by linkage studies. Analysis of tumor DNA from 42 patients shows that markers on chromosome 10 were lost in only one tumor. Thus, these results contrast with previous studies which show that tumor development is generally associated with the loss of the whole or substantial parts of the chromosome on which the putative tumor suppressor gene is located. PMID- 2565875 TI - Molecular structure of the human asparagine synthetase gene. AB - The human gene for asparagine synthetase has been isolated and the molecular organization has been determined by mapping and DNA sequencing of intron-exon boundaries. The gene spans 35 kb and contains 13 exons. The structure of the human gene has a high degree of similarity to that of the hamster asparagine synthetase gene, with identical positions for all but one of the intron regions. The 5' upstream region of this gene, like other housekeeping genes, lacks conventional TATA and CAAT boxes. Comparisons of the 5' upstream sequences of the human and hamster genes show limited similarity; however, both have a very high G + C content which may play a role in expression through DNA methylation. PMID- 2565876 TI - The human calbindin 27-kDa gene: structural organization of the 5' and 3' regions, chromosomal assignment, and restriction fragment length polymorphism. AB - The 5' and 3' regions of the human gene coding for calbindin 27 kDa were cloned and sequenced. Structural features of the 5' region included the presence of an Alu repeat and two elements regularly associated with eukaryotic promoters: an alternating purine-pyrimidine element and a homopurine-homopyrimidine box. The 3' region contained a second Alu family member and a degenerate 1.4-kb L1 repeat. A comparison with the chicken promoter was made in order to define regions conserved in evolution and potentially important in gene expression regulation. The greater similarity is located around the TATA box, but strongly conserved elements were not found. The gene was assigned to chromosome 8 by using human rodent hybrid cell lines. Two restriction fragment length polymorphisms (HindIII and SacI) were detected with a cDNA probe recognizing the 3' end of the gene. PMID- 2565877 TI - Linkage analysis of multiple endocrine neoplasia type 1 with INT2 and other markers on chromosome 11. AB - We evaluated linkage between the locus for multiple endocrine neoplasia type 1 (MEN1) and several polymorphic DNA markers on chromosome 11 in a single large pedigree. On the basis of the finding of a basic fibroblast growth factor (bFGF) like substance circulating in plasma of MEN1 patients, we chose a bFGF-related gene known to be localized to 11q13 as one of the markers. This gene locus, INT2, was found to be closely linked to the MEN1 gene. Pairwise and multipoint analyses with INT2 confirm the recent finding by C. Larsson et al. (1988, Nature (London) 332: 85-87) of MEN1 linkage to another marker, skeletal muscle glycogen phosphorylase, at 11q13. PMID- 2565879 TI - Localization of the gene for classic Alport syndrome. AB - The inheritance of Alport syndrome has been controversial for 30 years because no clear diagnostic criteria were established to define a clinically homogeneous group of patients. In this study, 41 families with "classic" Alport syndrome were identified and studied. All the pedigrees are compatible with X-linked inheritance. A formal genetic study confirmed linkage to probe S21 (DXS17), with a maximum LOD score of 4.72 at a recombination frequency of 0.06. PMID- 2565878 TI - Chromosomal localization of zinc finger protein genes in man and mouse. AB - We have determined the mouse and human chromosomal location of a gene (Zfp-3) that codes for a protein that contains potential DNA zinc-binding fingers. An analysis of the segregation of restriction fragment length polymorphisms in recombinant inbred strains and in an interspecific backcross demonstrated that Zfp-3 is located on mouse chromosome 11. Zfp-3 is very closely linked to the Trp53-1 locus but unlinked to another finger protein gene Zfp-4 located on mouse chromosome 8. In humans ZFP3 has been localized to chromosome 17p12-17pter and thus is part of the conserved linkage group between this chromosome and the distal half of mouse chromosome 11. PMID- 2565880 TI - The DNF15S2 locus at 3p21 is transcribed in normal lung and small cell lung cancer. AB - Small cell lung cancer (SCLC) has been associated with a deletion of the short arm of chromosome 3. One SCLC cell line, H748, has an interstitial deletion of chromosome 3p and shows allele loss for the DNF15S2 locus detected by the probe lambda H3. Conservation of DNF15S2 sequences in mouse indicated that this human genomic fragment may contain coding sequences. Screening of a normal lung cDNA library with chromosome 3-specific fragments of the lambda H3 probe resulted in the isolation of 18 positive clones. The cDNA clones detect an additional DNA polymorphism that is in linkage disequilibrium with the HindIII polymorphism of the DNF15S2 locus. Sequence analysis indicated that the DNF15S2 locus could potentially code for a previously unreported protein of 67 kDa which has 26 cysteine residues. DNF15S2 is part of the coding region of a 3.3-kb mRNA expressed in lung. Northern analysis indicated that this mRNA was not detectable in one of five SCLC lines. This SCLC line, H128, also lacks the enzyme aminoacylase 1. PMID- 2565881 TI - Localization of the human erbB-2 gene on normal and rearranged chromosomes 17 to bands q12-21.32. AB - Through the use of a cDNA probe, the human erbB-2 gene was localized by in situ hybridization of normal human chromosomes at 17q11-q21. In situ hybridization of chromosomes derived from fibroblasts carrying a constitutional 15;17t(q22.3;q11.21) translocation showed that the erbB-2 gene was relocated on the rearranged chromosome 15. These results as well as grain localization on prophase chromosomes locate the erbB-2 gene at 17q12-q21.32. This localization may facilitate the search for human malignancies with chromosome changes involving the erbB-2 gene. PMID- 2565882 TI - The necessity of both allogeneic antigens and stem cells for cyclophosphamide induced skin allograft tolerance in mice. AB - We have reported that in an H-2 identical murine combination of AKR/J (AKR, H-2k, Thy1.1) and C3H/HeJ (C3H, H-2k, Thy 1.2), specific tolerance to C3H skin in AKR mice is induced only when both intravenously (i.v.) 1 x 10(8) viable C3H spleen cells and, two days later, intraperitoneally (i.p.) 200 mg/kg cyclophosphamide (CP) have been given. To further examine this mechanism of tolerance, we used 2000R-irradiated C3H spleen cells as an antigen source and bone marrow cells depleted of Thy1.2+ cells and Ia+ cells as a stem cell source. When a mixture of 1 x 10(8) irradiated spleen cells and 3 x 10(7) bone marrow cells was used as tolerogen and 200 mg/kg CP was administered two days later, a profound and specific long-lasting tolerance was induced. This tolerant state, however, was less profound than that induced with spleen cells plus CP. When the number of irradiated spleen cells was fixed at 1 x 10(8), the tolerant state was dose dependent on the quantity of bone marrow cells. On the other hand, when the number of bone marrow cells was fixed at 1 x 10(6), tolerance induction depended on the dosage of irradiated spleen cells. Tolerance induced with irradiated spleen cells plus bone marrow cells and CP was tolerogen specific. Tolerance was never induced when the bone marrow cells had been irradiated with 2000R prior to injection. Transfer experiments showed that the tolerant state, in its acute phase, appeared to be predominantly based on reduction of functionally reactive cells. The prolongation of skin allograft survival in tolerant mice could not be attributed directly to suppressor cells, nor was any evidence of suppressive factor induction observed. In the chronic phase, however, the importance of the suppressive mechanisms appeared to be relatively increased. EPICS analysis of the thymocytes using fluorescein-conjugated anti-Thy1.1 and anti-Thy1.2 antibodies showed that a minimal degree of mixed chimerism had been established in the tolerant mice. Moreover, both T cells and Ia+ cells had beneficial effects on the induction of tolerance. We conclude that in the tolerance induced by spleen cells plus CP, histocompatibility antigens expressed on the surface of the spleen cells were essential to the antigen-stimulated cell destruction mechanism. Stem cells contained in the spleen cells also appeared to be crucial for maintaining tolerance by establishing a minimal degree of mixed chimerism. PMID- 2565883 TI - Immunoregulatory effects of cloned T cells on B-cell responses: comparison of autoimmune and non-autoimmune derived clones. AB - T-cell clones were derived from autoimmune prone (NZB) and non-autoimmune (C58) strains of mice and tested for their effects in several assays of B-cell responsiveness. Clones from the C58 strain suppressed lipopolysaccharide LPS stimulated B-cell proliferation, activation and immunoglobulin synthesis. In contrast, an NZB-derived clone enhanced these measures of B-cell response. The effects of the NZB clone were more notable on splenic target populations taken from mice 6 months of age or older. MHC-compatible DBA/2 spleen cells also showed enhancement of B-cell activation, but not of immunoglobulin synthesis by the NZB clone. It has been shown previously that all of the clones suppress T-cell proliferative responses. A potentially important skewing of the immune system toward humoral rather than cellular responses is therefore mediated by this clone derived from an autoimmune strain. PMID- 2565885 TI - Detection of individual interleukin 2-producing cells after anti-CD3 antibody activation. AB - A new intracytoplasmic immunofluorescence staining technique to detect and quantify human interleukin-2 (IL-2)-producing lymphocytes is described. Blood mononuclear cells (MNC) were cultured and stimulated by mitogenic anti-CD3 antibody to produce IL-2. The cells were then fixed and subsequently permeabilized in suspension by the detergent saponin. Cytoplasmic IL-2 could then be demonstrated using polyclonal IL-2 specific antibodies and indirect immunofluorescence staining. A characteristic morphology of the IL-2 staining was noted with a local circle-shaped accumulation in the cytoplasm in a perinuclear position, probably reflecting the presence of the lymphokine in the Golgi stacks. A rapid, but transient IL-2 production peak 6 h after initiation of the cultures was observed. Only 1% of the cells produced IL-2, but each IL-2 stained cell was very bright, indicating a low capacity of anti-CD3 antibody to induce IL-2 production rather than an insensitivity of our detection system. Of the IL-2 producing cells 90% were CD4-positive T cells and 10% were CD8-positive T cells as revealed by two-colour staining of cell surface antigens and cytoplasmic IL-2. PMID- 2565884 TI - T-cell receptor expression in intestinal intra-epithelial lymphocyte subpopulations of normal and athymic mice. AB - Intra-epithelial lymphocytes (IEL) in murine small intestine were analysed for the presence of cell-surface antigens and T-cell receptor allotype in normal and athymic BALB/c mice by immunoperoxidase histochemistry on frozen sections and immunofluorescence on isolated IEL. In frozen sections, IEL of normal mice were 97.7% CD45+, 93.5% CD3+, 46.2% Thy-1+, 91.1% CD8+, 10.7% CD4+ and 21.1% KJ16+ (V beta 8.1 and 8.2). FACS analysis of isolated IEL confirmed the level of KJ16 expression and also demonstrated that 25% of IEL were F23.1+ (V beta 8.1-8.3). Immunofluorescent double-staining revealed a skewed distribution of T-cell receptor (TcR) expression on Thy-1+ and Thy-1- IEL. KJ16 and F23.1 were expressed on 25.9% and 32.7% of Thy-1+ IEL, respectively; however, the frequency of V beta 8 expression was diminished on Thy-1- IEL (4.1% KJ16+ and 12.1% F23.1+). IEL are present in athymic mice, but at reduced levels. In frozen sections these cells were 91.9% CD45+, 69.5% CD3+, less than 1% Thy-1+, 83.6% CD8+, less than 1% CD4+ and less than 1% KJ16+. Thus it appears that in normal mice there may be two distinct lineages of IEL, a thymus-dependent Thy-1+ population which utilizes the alpha beta T-cell receptor and a thymus-independent Thy-1- population (represented in athymic mice), which may possibly utilize the alternative gamma delta TcR. PMID- 2565887 TI - The use of a monoclonal antibody against a proliferating cell nuclear matrix antigen in the study of solid human tumors. AB - A monoclonal antibody (MAb) identifies the nuclear antigen p125/6.5 associated with the nuclear matrix and present in proliferating human cells. By direct and indirect immunofluorescence, the presence and distribution of the antigen p125/6.5 in cryostat sections from primary and metastatic solid human tumors has been investigated. The antigen is present in nuclei of malignant and benign tumor cells, but is not detected in adjacent normal tissues. The antigen displays a speckled nucleoplasmic distribution, while nucleoli are negative. The intensity of the immunofluorescence reaction is markedly higher in the nuclei of some individual or grouped tumor cells. PMID- 2565886 TI - The polymorphic Fc gamma receptor II gene maps to human chromosome 1q. AB - Human receptors for IgG (Fc gamma R) play important roles in the immune response. Expression of the human Fc gamma RII gene may be relevant in immune complex related disorders such as systemic lupus erythematosus and Sjogren's syndrome. We have used spot blot analysis of dual laser-sorted human chromosomes to localize the Fc gamma RII gene to human chromosome 1. Spot blot analysis of sorted derivative chromosomes sublocalized the gene to the chromosome 1 long arm (1q12-- -q25.1). This subchromosomal localization involved reassigning a reciprocal chromosome translocation breakpoint. We also identified Xmn I and Taq I Fc gamma RII polymorphic restriction sites that arose before the races diverged. These common Xmn I and Taq I polymorphisms are predicted to be informative for segregation analysis with human diseases in 85% of all matings. PMID- 2565888 TI - Role of glucose-6-phosphate dehydrogenase on enhanced proliferation of pre neoplastic and neoplastic cells in rat liver induced by N-nitrosomorpholine. AB - The relationship between G6PD and the number and size of pre-neoplastic and neoplastic lesions in rat liver was examined. Male Sprague-Dawley rats were given drinking water containing 250 mg/l of NNM for 8 weeks. Pre-neoplastic and neoplastic lesions with positive staining of GT were classified histochemically into G6PD-positive and G6PD-negative lesions. At week 24, quantitative analysis showed that the incidence and number of G6PD-positive lesions were significantly higher than those of G6PD-negative lesions. Histologically, hepatocellular carcinomas were more frequent and larger in G6PD-positive lesions than in G6PD negative ones. Statistical analysis of the quantitative data also showed that the number and volume of G6PD-positive hepatocellular carcinomas were significantly higher than in G6PD-negative carcinomas. The G6PD-positive pre-neoplastic lesions had a significantly higher labelling index than G6PD-negative ones. These findings revealed a considerable difference in the growth of these 2 populations of the lesions, and indicate that G6PD activity may be closely related to proliferation of the pre-neoplastic and neoplastic cells in the liver. PMID- 2565889 TI - The biliary elimination of the selective beta-receptor blocking drug talinolol in man. AB - The biliary elimination of the beta-1-receptor blocking agent talinolol was investigated after intravenous administration of 30 mg in 6 patients with a T tube drain after cholecystectomy. Serum concentration decreased in a biexponential manner with a median terminal half-life time of 4.4 h (range: 3.0 6.2 h). In some patients a second peak of the serum level was found. Concentration-time curves in bile paralleled serum profiles. The bile:serum concentration ratio (b:s-ratio) ranged from 24 to 98. The biliary clearance amounted to 43 ml/h.kg (range: 13-212 ml/h.kg). The median of the amount of talinolol eliminated by bile was 2.8 mg (range: 1.1-7.4 mg). It is concluded that talinolol undergoes an enterohepatic circulation. However, the amount eliminated cannot provide a sufficient explanation for the second peak, observed in some patients. PMID- 2565890 TI - A democratic strategy for organizational change. AB - This article presents a model of democratic work organization based on concrete examples from Sweden. It focuses on how democratic work organizations can come about; how they can be introduced, developed, and protected; and, in general, how their growth can be encouraged. In the final section the perspective is broadened. The possibilities of creating an industrial democracy are discussed from a societal perspective. Women in the public sector are seen as a possible "avant-garde" in a union-based struggle for the democratic organization of work. PMID- 2565891 TI - Crystal-state structural analysis of two gamma-lactam-restricted analogs of Pro Leu-Gly-NH2. AB - The crystal structures of two analogs of Pro-Leu-Gly-NH2 (1), containing a gamma lactam conformational constraint in place of the -Leu-Gly- sequences, are described. The highly biologically active (S,R)-diastereomer 2a is semi-extended at the C-terminus, with the N-terminal Pro residue in the unusual "C5" conformation [psi 1 = -0.8(15) degrees] stabilized by a (peptide)N-H...N(amino) intramolecular H-bond [the N(3)...N(4) separation is 2.687(11)A]. Conversely, the N,N'-isopropylidene aminal trihydrate of the (S,S)-diastereomer 2b, compound 3, adopts a beta-bend conformation at the C-terminus, as already reported for 1. However, the backbone torsion angles [phi 2 = 57.4(4), psi 2 = -129.9(3) degrees; psi 3 = -92.3(4), phi 3 = 6.4(5) degrees] lie close to the values expected for the corner residues of an ideal type-II' beta-bend. A weak intramolecular 4----1 H-bond is seen between the Gly carboxyamide anti-NH and Pro C = O groups. In the newly formed 2,2,3,4-tetraalkyl-5-oxo-imidazolidin-1-yl moiety the psi 1 torsion angle is 12.9(4) degrees and the intramolecular N(3)...N(4) separation is 2.321(4)A. PMID- 2565892 TI - [Glomerulopathies in systemic diseases]. PMID- 2565893 TI - Vasoactive intestinal peptide (VIP)-like immunoreactivity in the human sympathetic ganglia. AB - Vasoactive intestinal peptide immunoreactive (VIP-IR) nerve fibres and terminals, neurons and small granule containing cells were observed in human lumbal sympathetic ganglia. Electron-microscopically VIP-IR was localized in the large dense-cored vesicles in nerve terminals and on the membranes of the Golgi complexes in the neurons. A small population of principal ganglion cells was surrounded by VIP-IR nerve terminals. Most of these neurons contained acetylcholinesterase (AChE) enzyme but were not tyrosine hydroxylase immunoreactive (TH-IR). All VIP-IR ganglion cells and most of the nerve fibres contained AChE but not TH-IR. It appears that in human sympathetic ganglia VIP is localized in the cholinergic neurons and nerve fibres and that the VIP-IR nerve terminals innervate mainly the cholinergic subpopulation of the sympathetic neurons. PMID- 2565894 TI - RFrAP: a computer program for analysis of complex HLA RFLPs. Assignment of restriction fragments to haplotypes by segregation analysis. AB - The availability of HLA class II cDNA probes has led to the development of a powerful method for the discrimination of genetic variation in this region of the major histocompatibility complex. There are problems with this approach which reduce its usefulness, including the hybridization of a probe with multiple restriction fragments (RFs) from the same locus or of two different cDNA probes with the same RF (cross-hybridizing). These problems may now be largely overcome by the relatively simple computer program presented here, which allows the entry, storage, and editing of phenotype data. It includes utilities for the automatic assignment of bands to haplotypes and also statistical functions to determine several types of correlations. The outputs are presented in a format familiar to HLA serologists and immunogeneticists. In addition, the data is stored in a manner that allows the user to easily prepare "ad hoc" figures and tables that illustrate the more complex relationships among bands and between them and other variables. PMID- 2565895 TI - The DR3(w18),DQw4 haplotype differs from DR3(w17),DQw2 haplotypes at multiple class II loci. AB - The polymorphism of HLA class II molecules in man is particularly evident when comparisons between population groups are made. This study describes a DR3 haplotype commonly present in the American black population. Unlike the Northern European population, in which almost all DR3 individuals are DQw2, approximately 50% of DR3-positive American blacks express a DQw4 allelic product. This study characterizes the DR subregion of that haplotype. cDNA sequence analysis has revealed a DR beta gene which differs at several positions from previously described DR3 beta 1 genes. It is postulated that a gene-conversion-like event with a DRw52 beta gene as donor has generated some of these differences. The haplotype carries a DRw52a allele as defined by oligonucleotide hybridization studies. DNA restriction fragment analysis using a family and several unrelated individuals has allowed us to identify DR alpha and beta fragments associated with the DR3(w18),DQw4 haplotype. The most striking observation is that the DR3(w18),DQw4 haplotype differs from DR3(w17),DQw2 haplotypes at multiple class II loci. Several genetic mechanisms including reciprocal recombination, gene conversion, and point mutation were involved in generating the differences between these haplotypes. Once established, the DR3(w18),DQw4 haplotype appears to be relatively stable in the population. PMID- 2565896 TI - Nobel lecture in physiology or medicine--1988. Drugs from emasculated hormones: the principle of syntopic antagonism. PMID- 2565897 TI - Patients with neuroleptic malignant syndrome histories: what happens when they are rehospitalized? AB - Among approximately 1450 patients treated with neuroleptics in a short-term psychiatric hospital, there were no cases of the neuroleptic malignant syndrome (NMS). Six patients with histories of NMS were treated, either with nonneuroleptic therapies or with low doses of low-potency antipsychotic agents. No case of NMS developed, but one patient suffered transient subsyndromal signs while treated briefly with loxapine. Staff education, screening for patients with history of NMS, and detection of early signs can lower the incidence of this serious drug toxicity. PMID- 2565899 TI - Enhanced presynaptic facilitation of vascular adrenergic neurotransmission in spontaneously hypertensive rats. AB - 1. The interaction between the vascular renin-angiotensin system and presynaptic beta-adrenoreceptors was examined in male and female normotensive and spontaneously hypertensive rats, using the in vitro perfused mesentery preparation. 2. Enhancement of the pressor response to periarterial nerve stimulation by isoprenaline was shown to be significantly greater in preparations from male and female spontaneously hypertensive rats compared to corresponding preparations from normotensive Wistar rats. 3. In preparations from normotensive and hypertensive animals, the potentiating effect of isoprenaline was prevented by pretreatment with propranolol or ICI 118 551, but not atenolol, implicating a beta 2-adrenoreceptor. 4. Angiotensin II enhanced the responses to peripheral nerve stimulation in preparation from normotensive and hypertensive animals. Enhancement was significantly greater in preparations from hypertensive animals. 5. The potentiation caused by isoprenaline was blocked by the angiotensin II receptor antagonist [Sar1-Ile8] angiotensin II, and by captopril. The potentiation following angiotensin II was unaffected by ICI 118 551. 6. These results suggest that stimulation of presynaptic beta 2-adrenoreceptors activates a localized angiotensin II system. No significant differences in this facilitatory system were observed between male and female animals, but the potentiation caused by activation of this system was considerably greater in the spontaneously hypertensive rats. PMID- 2565898 TI - Psychobiological effects of carbohydrates. AB - The authors studied whether the fatiguing effects of eating lunch are greater for carbohydrate-rich meals than for other meals, and related the time course of behavioral change to plasma glucose, insulin, and amino acids. On different occasions, in counterbalanced order, normal women (N = 7) fasted overnight, ate a standard breakfast, and at lunch either continued to fast or ate a high carbohydrate, low-protein meal; a hedonically similar meal containing both carbohydrate and protein; or a high-protein, low-carbohydrate meal. Meals were isocaloric and equated for fat content. Only the carbohydrate meal significantly increased fatigue, which could not be attributed to hypoglycemia because plasma glucose remained elevated. Fatigue began approximately, when the carbohydrate meal elevated the plasma tryptophan ratio but ended even though the ratio remained elevated. Fatigue after a high-carbohydrate lunch could not be explained by reactive hypoglycemia or sweet taste, and could partially be explained by the hypothesis that fatigue parallels an elevation of the tryptophan ratio. PMID- 2565900 TI - Presynaptic alpha 2-adrenoreceptors affecting terminal synaptic transmission by the nervi cardiaci accelerantes in the rat. AB - 1. Changes in heart rate were monitored in response to supramaximal electrical stimulation of the spinal sympathetic cardiac nerves in the pithed rat. 2. Submaximal positive chronotropic responses of the heart to stimulation (0.1-1.0 Hz) were inhibited after intravenous injection of the alpha 2-adrenoreceptor agonists B-HT 920 and xylazine, but not after the alpha 1-adrenoreceptor agonist methoxamine. 3. The alpha 2-adrenoreceptor antagonist yohimbine or idazoxan but not the alpha 1-adrenoreceptor antagonist prazosin, given intravenously, reduced the inhibitory effect of B-HT 920 on positive chronotropic responses to cardiac sympathetic nerve stimulation. Neither yohimbine, idazoxan nor prazosin had any effect on positive chronotropic responses to nerve stimulation. 4. Desmethylimipramine (DMI) potentiated the increased heart rate during continuous stimulation of the cardiac nerves at 0.2 Hz. Responses to stimulation, at this frequency, were unaffected by yohimbine, either in the absence or presence of the neuronal uptake inhibitor, DMI. 5. Intravenous infusion of the angiotensin antagonist saralasin or injection of the angiotensin converting enzyme inhibitor captopril had no significant effect, either on positive chronotropic responses to stimulation or on the inhibitory effects of B-HT 920. Positive chronotropic responses to stimulation were also unaffected by yohimbine in the presence of captopril, indomethacin, or indomethacin plus captopril. 6. Although presynaptic effects of alpha 2-agonists and their antagonists on cardiac nerve function were demonstrated, no evidence was obtained that prejunctional alpha 2-adrenoreceptors modulate transmission during continuous supramaximal firing at constant frequency.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565901 TI - A comparative study of gastric secretory stimulants in conscious dogs. AB - 1. In conscious dogs with gastric fistulae, Heidenhain pouches and Thomas duodenal fistulae, pentagastrin was found to be a more potent peak acid and pepsin stimulant in both innervated stomach and vagally denervated pouch than methacholine chloride. 2. Slopes of curves relating response to the logs of molar doses of pentagastrin and methacholine, at peak secretion, did not differ significantly. The maximal pentagastrin stimulated secretion from the pouch was smaller than that for methacholine; from the fistula they did not differ. 3. Ganglionic blockade depressed methacholine stimulated peak acid secretion, but did not affect pentagastrin stimulated acid secretion. Dose-response curves for methacholine-induced acid and pepsin secretion at the perigee did not differ from those obtained with ganglionic blockade. 4. Ganglionic blockade depressed pepsin secretion from the fistula whether stimulated with pentagastrin or methacholine. Pouch pepsin secretion was small and no difference between curves was seen. PMID- 2565902 TI - Failure of selective antagonists (CH-38083 and idazoxan) to distinguish between prejunctional and postjunctional alpha 2-adrenoreceptors. AB - 1. The prejunctional alpha 2-adrenoreceptor antagonist activity of CH-38083 (7,8 (methylenedioxi)-14-alpha-hydroxyalloberbane HCl), idazoxan and prazosin was determined against B-HT 920 on the tachycardia induced by cardiac nerve stimulation in pithed rats. Antagonism of cirazoline and B-HT 920 pressor responses was used to assess postjunctional alpha 1- and alpha 2-adrenoreceptor antagonist activities. 2. CH-38083 was more potent than idazoxan in blocking pre- and postjunctional alpha 2-adrenoreceptor sites. There was no difference between the activities of the two selective alpha 2-adrenoreceptor blocking agents on pre and postjunctional alpha 2-adrenoreceptors. 3. These data classify CH-38083 as a potent and highly selective alpha 2-adrenoreceptor antagonist in vivo and further support evidence of the homogeneity of pre- and postjunctional alpha 2 adrenoreceptors. PMID- 2565903 TI - A comparison of the properties of prenalterol and corwin at beta 1- and beta 2 adrenoreceptors in vitro. AB - 1. The affinities and efficacies (relative to isoprenaline) of prenalterol and corwin at beta 1- and beta 2-adrenoreceptors, have been determined in isolated cardiac and vascular tissues respectively. 2. Prenalterol and corwin have similar affinities for cardiac beta 1-adrenoreceptors. The affinity of prenalterol for beta 2-adrenoreceptors is approximately 10 times lower than for beta 1 adrenoreceptors; that for corwin is approximately 100 times lower than for beta 1 adrenoreceptors. 3. The efficacies of prenalterol and corwin, relative to isoprenaline, at beta 1 and beta 2-adrenoreceptors, are similar. 4. The greater selectivity of corwin compared with prenalterol, as an agonist at beta 1 adrenoreceptors, is a reflection of its lower affinity for beta 2 adrenoreceptors. PMID- 2565904 TI - Coat proteins isolated from clathrin coated vesicles can assemble into coated pits. AB - Isolated human fibroblast plasma membranes that were attached by their extracellular surface to a solid substratum contained numerous clathrin coated pits that could be removed with a high pH buffer (Moore, M.S., D.T. Mahaffey, F.M. Brodsky, and R.G.W. Anderson. 1987. Science [Wash. DC]. 236:558-563). When these membranes were incubated with coat proteins extracted from purified bovine coated vesicles, new coated pits formed that were indistinguishable from native coated pits. Assembly was dependent on the concentration of coat protein with half maximal assembly occurring at 7 micrograms/ml. Assembly was only slightly affected by the presence of divalent cations. Whereas normal appearing lattices formed in a low ionic strength buffer, when assembly was carried out in a low pH buffer, few coated pits were evident but numerous small clathrin cages decorated the membrane. Coated pits did not form randomly on the surface; instead, they assembled at differentiated regions of membrane that could be distinguished in carbon/platinum replicas of frozen and etched membranes by the presence of numerous particles clustered into patches the size and shape of a coated pit. PMID- 2565905 TI - The propeptide of preprosomatostatin mediates intracellular transport and secretion of alpha-globin from mammalian cells. AB - We have investigated the role of the somatostatin propeptide in mediating intracellular transport and sorting to the regulated secretory pathway. Using a retroviral expression vector, two fusion proteins were expressed in rat pituitary (GH3) cells: a control protein consisting of the beta-lactamase signal peptide fused to chimpanzee alpha-globin (142 amino acids); and a chimera of the somatostatin signal peptide and proregion (82 amino acids) fused to alpha-globin. Control globin was translocated into the endoplasmic reticulum as determined by accurate cleavage of its signal peptide; however, alpha-globin was not secreted but was rapidly and quantitatively degraded intracellularly with a t 1/2 of 4-5 min. Globin degradation was insensitive to chloroquine, a drug which inhibits lysosomal proteases, but was inhibited at 16 degrees C suggesting proteolysis occurred during transport to the cis-Golgi apparatus. In contrast to the control globin, approximately 30% of the somatostatin propeptide-globin fusion protein was transported to the distal elements of the Golgi apparatus where it was endoproteolytically processed. Processing of the chimera occurred in an acidic intracellular compartment since cleavage was inhibited by 25 microM chloroquine. 60% of the transported chimera was cleaved at the Arg-Lys processing site in native prosomatostatin yielding "mature" alpha-globin. Most significantly, approximately 50% of processed alpha-globin was sorted to the regulated pathway and secreted in response to 8-Br-cAMP. We conclude that the somatostatin propeptide mediated transport of alpha-globin from the endoplasmic reticulum to the trans-Golgi network by protecting molecules from degradation and in addition, facilitated packaging of alpha-globin into vesicles whose secretion was stimulated by cAMP. PMID- 2565907 TI - Differentiation of germ cells in seminiferous tubules transplanted to testes of germ cell-deficient mice of W/Wv and Sl/Sld genotypes. AB - (WB X C57BL/6)F1-W/Wv (hereafter, WBB6F1-W/Wv) mice and (WC X C57BL/6)F1-Sl/Sld (hereafter, WCB6F1-Sl/Sld) mice are sterile due to the deficient spermatogenesis in the testes. The cause of deficient spermatogenesis in WBB6F1-W/Wv mice is considered to be a defect in germ cells themselves, whereas that in WCB6F1-Sl/Sld mice is considered to be a defect in tissue environment necessary for differentiation of germ cells. Seminiferous tubules isolated from cryptorchid testes of C57BL/6- +/+ mice were transplanted into the testes of WBB6F1-W/Wv and WCB6F1-Sl/Sld mice to clarify that the extratubular environment of these mice was intact or not. Type A spermatogonia in the transplanted tubules normally differentiated into spermatids, suggesting that the extratubular environment is intact in both WBB6F1-W/Wv and WCB6F1-Sl/Sld mice. PMID- 2565906 TI - Ciliary neurotrophic factor induces cholinergic differentiation of rat sympathetic neurons in culture. AB - Ciliary neurotrophic factor (CNTF) influences the levels of choline acetyltransferase (ChAT) and tyrosine hydroxylase (TH) in cultures of dissociated sympathetic neurons from newborn rats. In the presence of CNTF both the total and specific activity of ChAT was increased 7 d after culture by 15- and 18-fold, respectively, as compared to cultures kept in the absence of CNTF. Between 3 and 21 d in culture in the presence of CNTF the total ChAT activity increased by a factor of greater than 100. Immunotitration demonstrated that the elevated ChAT levels were due to an increased number of enzyme molecules. In contrast to the increase in ChAT levels, the total and specific activity levels of TH were decreased by 42 and 36%, respectively, after 7 d in culture. Half-maximal effects for both ChAT increase and TH decrease were obtained at CNTF concentrations of approximately 0.6 ng and maximal levels were reached at 1 ng of CNTF per milliliter of medium. The effect of CNTF on TH and ChAT levels were seen in serum containing medium as well as in serum-free medium. CNTF was shown to have only a small effect on the long-term survival of rat sympathetic neurons. We therefore concluded that the effects of CNTF on ChAT and TH are not due to selective survival of cells that acquire cholinergic traits in vitro, but are rather due to the induction of cholinergic differentiation of noradrenergic sympathetic neurons. PMID- 2565908 TI - Retroviruses expressing different levels of the normal epidermal growth factor receptor: biological properties and new bioassay. AB - Two retroviral DNAs that encode the normal human epidermal growth factor (EGF) receptor hEGFR have been generated by inserting a hEGFR cDNA into two different retroviral vectors. One DNA (pCO11-EGFR-neo) also contained a linked selectable marker gene (neoR). The other (pCO12-EGFR) only expresses hEGFR. When introduced into NIH3T3 cells, the two DNAs and the viruses derived from them induced a fully transformed phenotype, including focal transformation and growth in agar or low serum, but transformation depended entirely upon EGF being present in the growth medium. Compared with pCO11-EGFR-neo, pCO12-EGFR induced EGF-dependent transformation 2-5 times more efficiently and expressed higher numbers of receptors (4 x 10(5) vs. 1 x 10(5) EGF receptors per cell). The results indicate that transforming potential is directly related to the number of EGF receptors. In defined, serum-free medium that contained only very low concentrations of insulin (0.6 microgram/ml) and transferrin (0.6 micrograms/ml), hEGFR-virus infected cells were able to grow with EGF as the only growth factor. Moreover, daily incubation of the cells with EGF for only 30 min was sufficient to induce growth. NR6 cells, which lack endogenous EGF receptors, were transformed as efficiently as NIH3T3 cells by the hEGFR virus. The dose-dependent growth response to EGF of infected NR6 cells grown in serum-free medium can be used as a highly sensitive bioassay for the quantitative assessment of EGF and transforming growth factor type alpha (TGF alpha). This bioassay is at least as sensitive as previously reported radioimmunoassays and can measure a much wider concentration range (10 pg-100 ng/ml). Uninfected NR6 cells or NR6 cells infected by helper virus alone can be used as controls for the EGF specificity of growth stimulation. PMID- 2565909 TI - Overexpression of the c-erbB-2 protein in human breast tumor cell lines. AB - The c-erbB-2 proto-oncogene is amplified in a high percentage of primary human breast tumors, suggesting that the overexpression of this gene may be involved in the development of human breast cancer. We have investigated five human breast tumor cell lines and have detected amplified c-erbB-2 gene copies in two of them. This amplification leads to overexpression of the c-erbB-2 protein. In addition, two other cell lines have elevated protein levels without gene amplification, suggesting that other mechanisms can lead to overexpression of the c-erbB-2 protein. These results are similar to those that we obtained during a study of primary breast tumors (Berger et al.: Cancer Res 48:1238-1243, 1988). These breast tumor cell lines should be useful for an analysis of c-erbB-2 expression and of the mechanisms that in some cases lead to overexpression. PMID- 2565910 TI - The metabolic centroid method for PET brain image analysis. AB - The method of centroids is an approach to the analysis of three-dimensional whole brain positron emission tomography (PET) metabolic images. It utilizes the brain's geometric centroid and metabolic centroid so as to objectively characterize the central tendency of the distribution of metabolic activity in the brain. The method characterizes the three-dimensional PET metabolic image in terms of four parameters: the coordinates of the metabolic centroid and the mean metabolic rate of the whole brain. These parameters are not sensitive to spatially uniform random noise or to the position of the subject's head within a uniform PET camera field of view. The method has been applied to 40 normal subjects, 22 schizophrenics who were treated with neuroleptics, and 20 schizophrenics who were neuroleptic-free. The mean metabolic centroid of the normal subjects was found to be superior to the mean geometric centroid of the brain. The mean metabolic centroid of chronic schizophrenics is lower and more posterior to the mean geometric centroid than is that of normals. This difference is greater in medicated than in unmedicated schizophrenics. The posterior and downward displacement of the mean metabolic centroid is consistent with the concepts of hypofrontality, hyperactivity of subcortical structures, and neuroleptic effect in schizophrenics. PMID- 2565911 TI - Isolation, separation and analysis in neurochemistry: trace amines and acids as an illustrative example. AB - In this brief overview various neurochemical isolation procedures that can be adopted for the analysis of several monoamine neurotransmitters/neuromodulators and their principal oxidatively deaminated metabolites are outlined. With respect to the trace amines, they can be identified and quantitated as their so-called dansyl derivatives after thin-layer chromatographic separation by mass spectrometric (MS) electron-impact (EI) ionisation followed by selected-ion monitoring (SIM) of their molecular ions. Deuterated homologues are added as internal standards at the start of the analytical procedure. The MS-EI-SIM procedure offers a tissue extract or releasate sensitivity of about 100 pg/g of tissue or fluid. In the case of tryptamine or phenylethylamine, by utilising different derivatives (N-acetylpentafluoropropionyl or N acetylpentafluorobenzoyl), which cyclise to form perfluorinated spirocyclic compounds, it is possible using MS negative chemical ionisation techniques coupled with monitoring of the (M-HF) ions to achieve sensitivities for tissue extracts of 1 pg/g or less. Acidic and neutral metabolites (up to twelve of them can be assayed simultaneously) can be detected and quantitated in tissue extracts, releasates or biological fluids as their methylpentafluoropropionyl or trifluoroethyl-pentafluoropropionyl derivatives in the 100-1000 pg range using gas chromatographic-MS-SIM procedures. PMID- 2565912 TI - Dose-response study and long term effect of the somatostatin analog octreotide in patients with therapy-resistant acromegaly. AB - Twelve acromegalic patients in whom standard therapy was unsuccessful were evaluated with 24-h serum GH profiles (hourly sampling) and oral glucose tests (oGTT) while being treated with octreotide, a long-acting somatostatin analog. During a dose-response study (300, 600, and 1500 micrograms/day sc, for 4 weeks), serum GH decreased significantly after 300 micrograms/day in 8 of 12 patients [from 14.5 +/- 6.2 (+/- SE) to 4.9 +/- 1.9 micrograms/L]. Higher doses further reduced serum GH concentrations in 3 (600 micrograms/day) and 1 (1500 micrograms/day) patients, respectively. Four patients did not respond to any dose. Serum GH concentrations declined normally (GH nadir, less than 2 micrograms/L) after glucose ingestion in 4 of the 10 nondiabetic acromegalic patients. In 4 patients, including 2 of the initial nonresponders, serum GH further declined during long term treatment (12 and 18 months). In the latter 2 patients, serum insulin-like growth factor I (IGF-I) concentrations had decreased during the dose-response study despite the absence of measurable GH suppression. Eight patients attained normal serum IGF-I concentrations during treatment. Serum IGF-I and GH correlated significantly before, but not during, treatment. Retrospective comparison suggested that in 5 of 6 patients, serum GH was more effectively suppressed by octreotide than by bromocriptine. The 24-h serum octreotide concentration varied greatly among the patients. Although the 24-h serum octreotide and GH concentrations did not correlate with one another, the serum octreotide and IGF-I concentrations when the patients were receiving 300 micrograms/day tended to be negatively correlated (r = -0.496; P = 0.118). The 24 h serum insulin values decreased and those of glucose increased during treatment; after oral glucose, serum insulin was lower and glucose was higher. However, after 12 months of treatment, the 8-h serum insulin profile and peak serum insulin after oral glucose administration had returned to pretreatment values, while serum glucose remained abnormal. We conclude that 1) octreotide lowers serum GH in many, but not all, acromegalic patients resistant to other forms of treatment; 2) doses in excess of 300 micrograms/day should be tested in those patients in whom lower doses are ineffective; 3) serum IGF-I measurement may be a better indicator of treatment success than GH measurement; 4) octreotide concentrations do not correlate with GH suppression; and 5) deterioration of carbohydrate tolerance does occur but tends to improve during chronic treatment. PMID- 2565913 TI - Long term effects of continuous subcutaneous infusion of the somatostatin analog octreotide in the treatment of acromegaly. AB - The marked pituitary tumor shrinkage achieved by continuous sc infusion (CSI) of the long-acting somatostatin analog octreotide in one acromegalic patient led us to treat 16 other acromegalic patients for up to 24 months by CSI. This therapy, given in doses ranging from 100-600 micrograms/day, resulted in normalization of the mean daily serum GH (mGH) and insulin-like growth factor I levels in 9 of the 17 patients (53%). In 7 patients, mean daily serum GH decreased but not to normal; 3 of these patients had hyperprolactinemia which was not influenced by octreotide. One patient was completely unresponsive. In contrast to the biochemical results, 80% of the patients had marked clinical improvement. Side effects consisted of slightly impaired carbohydrate tolerance in 2 patients and cholelithiasis in 2 patients. Pituitary tumor size decreased in only 3 patients; in 1 of them visual field defects disappeared rapidly. These results suggest that octreotide treatment may prove beneficial before surgery in patients with macroadenomas, although its efficacy varies widely. Potential responsivity can usually be determined by a short course (24 h) of CSI of octreotide. PMID- 2565914 TI - Inhibition of the growth hormone (GH) response to GH-releasing hormone by constant Met-GH infusions. AB - GH release is controlled by hypothalamic hormones and insulin-like growth factor I, synthesized under the influence of GH, and perhaps also by GH itself. The availability of recombinant Met-GH was the basis for studies aimed at 1) obtaining constant serum GH levels by means of constant Met-GH infusions (40 and 80 ng/kg.min for 6 h), and 2) evaluating the metabolic effects of constant GH levels and, in particular, their effects on the serum GH response to GHRH. In six normal men, both Met-GH infusions increased plasma FFA levels, but did not alter the circulating levels of somatostatin, insulin-like growth factor I, insulin, glucose, cholesterol, and triglycerides. The Met-GH infusions did cause a dose related inhibition of GHRH-induced GH release. These data indicate that it is possible to maintain constant serum GH levels by means of constant Met-GH infusions at different infusion rates, and that GH inhibits its own release. PMID- 2565915 TI - Suppression of ectopic adrenocorticotropin secretion by the long-acting somatostatin analog octreotide. AB - The long-acting somatostatin analog (octreotide) was administered to a 37-yr-old woman with the ectopic ACTH syndrome. The patient had diffuse metastatic spread of a nonpituitary tumor, presumably of pancreatic origin, and severe and rapidly progressive hypercortisolism with extreme myopathy, hypokalemia, and diabetes mellitus. Plasma ACTH and lipotropin levels and 24-h urinary cortisol excretion were greatly elevated [218 pg/mL (48 pmol/L), 1340 pg/mL (220 pmol/L), and up to 830 micrograms/24 h (2290 nmol/day), respectively]. Urinary cortisol excretion decreased to normal within 3 days after the initiation of octreotide therapy (150, 300, and 600 micrograms/day), and plasma ACTH and lipotropin levels also decreased. Urinary cortisol excretion remained normal for 2 months during chronic octreotide therapy, and her general condition improved dramatically. The only side-effect was a slight increase in the number of bowel movements. Tumor progression, however, was not controlled, and she eventually died of hepatic insufficiency. These data indicate that octreotide can be a highly effective treatment for patients with the ectopic ACTH syndrome. PMID- 2565916 TI - Immunoreactive somatostatin in male reproductive system in humans. AB - Somatostatin-like immunoreactivity is distributed widely in humans; the highest concentration is in the central nervous system, gastrointestinal tract, and pancreas. To determine if somatostatin is present in the male reproductive system, we analyzed human testis, epididymis, prostate, and semen. Somatostatin like immunoreactivity was detectable in acid extracts of human testis, epididymis, and prostate (n = 6 each) in concentrations of 4.0 +/- 1.4 (+/- SD), 14.7 +/- 3.2, and 27.5 +/- 5.1 pmol/g wet wt, respectively. Considerable amounts of immunoreactive somatostatin also were detectable in semen; the mean value was 3.8 +/- 1.3 nmol/L (n = 6). This value was 200-fold higher than that in peripheral plasma. The somatostatin immunoreactivity in these tissues was characterized by gel filtration chromatography. Two peaks of somatostatin immunoreactivity, one coeluting with somatostatin-14 and the other with somatostatin-28, were found in the testis, epididymis, prostate, and hypothalamus. The amounts of the two sizes were nearly equal in the testis; somatostatin-14 predominated in the epididymis, prostate, and hypothalamus; whereas only somatostatin-28 was detected in semen. The presence of somatostatin in the male reproductive system suggests that somatostatin may play a role in the regulation of reproductive function in men. PMID- 2565917 TI - Alpha-1 adrenoceptor blockade with doxazosin in hypertension: effects on blood pressure and lipoproteins. AB - The effects of doxazosin, a long-acting alpha-1 adrenoreceptor blocking drug, were observed upon blood pressure and serum lipoproteins. Thirty patients with supine diastolic blood pressure between 90 and 114 mm Hg during single-blind placebo therapy were randomized to double-blind treatment with either doxazosin or further placebo in a parallel-design protocol. Starting at one mg, dosage was doubled every 2 weeks during a 10-week treatment period to a maximum dose of 16 mg once daily. Blood was sampled in the fasting state before and during double blind therapy for measurement of total cholesterol and triglycerides, cholesterol in the lipoprotein fractions, and apolipoproteins A and B. At the end of 10 weeks of titration, systolic and diastolic blood pressure were each reduced by 14 mm Hg in the standing position when measured 24 hours following the previous dose. Supine pressure was lowered by 6 mm Hg systolic and by 5 mm Hg diastolic at the same time point. Measured hourly for 12 hours following the ingestion of doxazosin, blood pressure was lowered maximally at 4-5 hours when an additional decline of 14/6 mm Hg (systolic/diastolic) was observed in the standing position and 13/6 in the supine posture. Postural dizziness, the most frequent symptomatic complaint, was reported in 4 patients during doxazosin treatment. After brief interruption of treatment in one and dosage adjustment in another, titration was continued in all four and no patient was withdrawn because of side effects. Concerning lipoproteins, the ratio of total cholesterol to HDL cholesterol and of LDL to HDL cholesterol both improved during treatment with doxazosin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565918 TI - Bronchial and cardiovascular effects of ocular topical B-antagonists in asthmatic subjects: comparison of timolol, carteolol, and metipranolol. AB - B antagonists eye drops are most effective for the treatment of chronic open angle glaucoma. By this way of administration they have a very good systemic bioavailability. Bronchial, and cardiovascular effects of three of these topicals: timolol, carteolol and metipranolol have been evaluated in three parallel groups of asthmatic patients. The three topics induce bronchoconstriction without significant difference between them, and lower heart rate (sometimes very intensely) whatever the B antagonist studied. From these data, it is recommended to practitioners to follow carefully the rules of administration of B blockers, even in eye drops. PMID- 2565919 TI - Isolation and purification of scrapie-associated fibrils and prion protein from scrapie-infected hamster brain. AB - We report the purification of prion protein (PrP) 27-30 and scrapie-associated fibrils (SAF) from hamsters infected with the 263K strain of scrapie. SDS-PAGE of fractions purified from scrapie-infected brains revealed several bands at approximately 28.5 kDa, 23.9 kDa and 14.3 kDa and, in one set of preparations, a protein of Mr 26 kDa was found in both scrapie-infected and sham-inoculated animals. The specificity of PrPs was confirmed by Western blotting. Ultrastructural analysis of fractions from scrapie-infected brains revealed numerous fibrils measuring approximately 20 nm in diameter and 100 to 200 nm in length. The substructure of these fibrils consisted of protofilaments which were usually straight and rarely helically arranged. We conclude that the electron microscopical appearance of SAF depends much on the purification scheme. The PrP27-30 as well as proteins of lower Mr are easily detectable in scrapie infected brains. The detection of protein of a Mr 26 kDa in both scrapie-infected and sham-inoculated animals suggests that this form of PrP may exist in equilibrium with PrP33-35c. PMID- 2565920 TI - Traumatic plantar urticaria or plantar erythema nodosum. PMID- 2565921 TI - Sulfasalazine: a potential psoriasis therapy? AB - In an open study, 32 patients with moderate to severe stable plaque psoriasis were treated with sulfasalazine, 3 gm daily for 8 weeks. Twenty-four patients completed the study, and 19 had modest to marked improvement or clearing. If confirmed by a double-blind study, sulfasalazine could become an alternative therapy in some patients with moderate to severe psoriasis. PMID- 2565922 TI - The histochemical location of three diagnostic enzymes in the marmoset liver. AB - The histochemical locations of alkaline phosphate (ALP), leucine aminopeptidase (LAP) and gamma glutamyltransferase (GGT) were demonstrated in the liver of the marmoset (Callithrix jacchus). Although all three enzymes were located in cell membranes, the location of LAP was demonstrated by a chromogenic substrate, in the canalicular membrane. GGT was seen in a vascular network, provisionally identified as the peribiliary arterial plexus. Possible diagnostic applications in toxicology are discussed. PMID- 2565923 TI - Stingray spine foreign body in the foot. AB - An unusual case report of a retained stingray spine foreign body in the foot is presented. The case is interesting in that the foreign body, although visualized on radiograph, was difficult to locate intraoperatively. A review of techniques for localizing foreign bodies in the foot is presented as well as a discussion on stingray injuries in general. Because of the possibility of spine breakage in a stingray puncture wound, radiographs should be considered in injuries of this type. PMID- 2565924 TI - Food restriction prevents an age-associated increase in rat liver beta-adrenergic receptors. AB - In male Wistar rats fed ad libitum (24% protein, 4.5 Kcal/gm), the [125I]iodopindolol binding capacity of the beta-adrenergic receptors in liver of 24-month-old animals is 3-4 times greater than that of 6-month-old counterparts. In rats fed the same diet, on alternate days from weaning, the receptor capacity did not increase significantly between 6 and 24 months (10.20 +/- 0.55 vs 9.20 +/ 0.72 fmol/mg) or between 24 and 30 months. This was not due to acute dietary deprivation, as rats food-restricted for only 2 weeks, at 23.5 months of age, also showed elevated receptor capacities compared to 6-month-old ad libitum fed animals. Moreover, intermittent feeding produced no significant effects among 6 month-old animals, whether restricted since weaning or for two weeks prior to sacrifice. Many biochemical parameters that decrease with aging in rats fed ad libitum are prevented by dietary restriction. Our results demonstrate that a reproducible biochemical process that increases with aging is also prevented with dietary restriction. The age-related, liver beta-receptor increase may be a potentially reliable marker for studying biochemical perturbations that modify life span. PMID- 2565925 TI - Colchicine prevents D-galactosamine-induced hepatitis. AB - The hepatoprotective effect of colchicine in a model of liver intoxication with galactosamine (GalN), 375 mg/kg, i.p., was studied in rats. At 0.5, 1, 3, 6, 18 and 24 h after GalN intoxication the following markers of liver damage were measured: serum activity of alanine aminotransferase, alkaline phosphatase, gamma glutamyltranspeptidase, hepatic calcium and glycogen contents, liver lipoperoxidation, and liver plasma membrane activity of alkaline phosphatase, gamma-glutamyltranspeptidase and high-affinity Ca2+-ATPase. 24 h after GalN intoxication increases in serum levels of alanine aminotransferase, alkaline phosphatase and gamma-glutamyltranspeptidase were observed along with decreases in plasma membrane activities of alkaline phosphatase, gamma glutamyltranspeptidase, and high-affinity Ca2+-ATPase. A sharp increase of lipoperoxidative processes measured as malondialdehyde production was also observed. Pretreatment of rats with colchicine 10 micrograms/rat/day p.o. for 7 days before GalN injection prevented partially the toxic effects of GalN. When a dose of 50 micrograms/rat/day for 7 days was given the drug prevented almost completely the damage induced by galactosamine, with the exception of glycogen and serum alkaline phosphatase that remained different from controls. Time-course experiments showed that malondialdehyde formation increased 30 min after intoxication while all other changes became apparent from 6 h after treatment, suggesting that lipoperoxidation may be a prerequisite for galactosamine-induced damage. The protection offered by colchicine was related to its capacity to inhibit lipoperoxidation. Histochemical findings paralleled the biochemical results. The possible role of lipoperoxidation in galactosamine-induced liver damage is discussed. PMID- 2565926 TI - The cell surface glycoprotein Mac-1 (CD11b/CD18) mediates neutrophil adhesion and modulates degranulation independently of its quantitative cell surface expression. AB - It has previously been shown that during degranulation Mac-1 (CD11b/CD18)--a glycoprotein that plays a central role in neutrophil adhesion-is up-regulated on PMN surfaces. It has been assumed that this quantitative change in adhesion Ag expression on the cell surface would in turn lead to increased cellular adhesiveness. In contrast, we found that at an incubation temperature of 16 degrees C, stimulated neutrophil adhesion to plastic tissue culture dishes in the presence of FMLP (2.5 x 10(-6) M), TNF (10 ng/ml), or PAF (1 x 10(-4) M) occurred without cellular degranulation or Mac-1 surface up-regulation as measured cytofluorometrically. As shown by functional inhibition studies employing monoclonal antibodies 60.3 (anti-CD18) and 60.1 (anti-CD11b), adhesion at 16 degrees C, where no CD11b/CD18 up-regulation was seen, is mediated by CD11b/CD18 just as it is at 37 degrees C, where degranulation and CD11b/CD18 up-regulation could be demonstrated. The physiologic importance of these findings was underscored by experiments done on endothelial monolayers, which showed that PMN association with endothelial cells is absolutely independent from the quantitative up-regulation of Mac-1 on PMN surfaces. When neutrophils were stimulated at 37 degrees C by endotoxin, an agent that does not induce aggregation (a form of intercellular adhesion), Mac-1 surface expression increased only after cells had become adherent, whereas cells held in suspension to prevent cell-substrate adhesion neither degranulated nor up-regulated their Mac-1 surface expression. Thus, not only is adherence independent of degranulation and Mac-1 cell surface up-regulation, but both degranulation and Mac-1 surface up-regulation appear to depend on the process of adhesion. Correspondingly, incubation of neutrophils with antibodies 60.1 and 60.3 inhibited not only adhesion of cells stimulated with FMLP at 37 degrees C but degranulation as well. These results indicate that Mac-1 influences degranulation as well as it controls adhesion not by its mere quantity on the cell surface, but rather by an yet undefined molecular modulation. PMID- 2565927 TI - Variability of CR2 gene products is due to alternative exon usage and different CR2 alleles. AB - The gene encoding human complement receptor 2 (CR2) lies within a cluster of genes on human chromosome 1. Polymorphisms have been described for several of these genes or gene products. To examine any possible polymorphisms in the CR2 gene product we have analyzed the CR2 transcriptional products for variations via alternative exon usage and for the presence of different CR2 alleles within the human population. This analysis has suggested that an exon encoding a 60 amino acid short consensus repeat can be alternatively spliced. Evidence for two transcriptional products has been found in a variety of transformed human B cells and two human tonsils. Interestingly the ratio of the two forms of mRNA, that which includes the alternative exon and that which does not, is not constant when examining the RNA from these sources. In addition to the variation introduced by the alternatively spliced product, there appears to exist in the human population at least three different alleles for CR2. These alleles have been defined by multiple nucleotide changes which are not silent but which alter the amino acid encoded at that site. However, we have not identified allelic variants of CR2 which would produce a product varying in the number of long homologous repeats, as has been identified for the various alleles of the closely related protein, human complement receptor 1. PMID- 2565928 TI - Numbers and functions of transplantable primitive immunohematopoietic stem cells. Effects of age. AB - This report introduces a new method in immunology, a use of the binomial formula with covariance to estimate numbers and proliferative patterns of the most primitive lymphoid precursors. We studied the primitive stem cells (PSC) from which most circulating lymphocytes and erythrocytes were descended during 300 to 400 days in recipients of genetically distinguishable marrow mixtures in competitive repopulation. Equivalent PSC concentrations (Eq. PSC Conc. or Conc.) were estimated, with the notion of common PSCs contributing equally in lymphoid and myeloid compartments. Similar estimation was done for common PSCs from which lymphocytes (and erythrocytes) drawn at successive sampling times about 100 days apart were descended. The percentages of lymphocyte and erythrocyte types, P1 and Pe, measured in each recipient were closely correlated, especially after 6 months and later. Close correlations were also found in cells sampled at successive one hundred day intervals, especially after the first. Apparently a few PSCs or their direct descendents produced most of the blood lymphocytes and E, and this production continued for many months. Concentrations of these PSCs (Equivalent PSC concentrations) were about one per 10(5) marrow cells from young donors. This is much lower than previous estimates, probably because our methods focus only on the most interesting precursors, those from which most of the circulating cells were descended. Equivalent PSC concentrations were about two-fold higher in old donors; old marrow produced correspondingly higher P1 and Pe values, but these declined with time. There were also small increases with time in the P1 and Pe values with young donors. To explain the temporal trends, we suggest that excess concentrations of precursors less primitive than PSC are present in old marrow, and their contribution to the differentiated cell population gradually declines. Possibly such precursors, as well as true PSC, proliferate in old donors to compensate for deficiencies that develop with age. PMID- 2565930 TI - Activation of human cytolytic cells through CD2/T11. Comparison of the requirements for the induction and direction of lysis of tumor targets by T cells and NK cells. AB - We studied the mechanisms whereby human T cells and NK cells are activated and directed to lyse tumor targets through the CD2 (T11/E-rosette) Ag. Using two cloned NK lines, we showed that these cells, as had previously been shown for T cells, could be directed to lyse an "NK-resistant" tumor target in the presence of antibody heterodimers. These heterodimers consisted of a (mAb) to CD2 (anti T11(2) or anti-T11(3] linked to a mAb recognizing the tumor cell (J5, anti CALLA). However, distinct differences between NK cells and T cells were observed with regard to the requirements for such directed lysis: first, only one epitope of CD2 on NK cells (either T11(2) or T11(3] needed to be recognized by the antibody heterodimer in order for directed lysis to occur, whereas for T cells both T11(2) and T11(3) epitopes had to be recognized. Second, in confirmation of previous data with monomeric anti-T11(2) or anti-T11(3) antibody, heterodimers constructed with these reagents enhanced conjugate formation between NK cells and tumor targets, whereas no such enhancement was seen with T cells. All types of heterodimer directed lysis were dependent on the adhesion molecule LFA-1, as an anti-LFA-1 antibody-blocked lysis. Third, whereas in T cells lysis mediated through CD2 appeared to be regulated by CD3 but not vice versa, all types of lysis by NK cells appeared to be regulated through CD2. Finally we showed that F(ab')2 fragments of the anti-T11(2) and anti-T11(3) antibodies could activate NK cells, but were unable to activate T cells either as cloned cytolytic lines, or in populations of PBL. The implications of our findings with regard to the role of CD2 in the activation of cytolytic cells is discussed. PMID- 2565929 TI - Requirement of leukocytic cell adhesion molecules (CD11a-c/CD18) in the enhanced NK lysis of iC3b-opsonized targets. AB - Treatment of Raji or Daudi cells with human serum under conditions which allow the alternative pathway of C activation results in their C3-opsonization and enhanced sensitivity to NK-mediated lysis. The effector lymphocytes have low buoyant density, carry CD16 and HNK1 markers as well as the CD11a-c/CD18 leukocytic cell adhesion molecules. One of these molecules, made up of CD11b-CD18 (alpha- and beta-chains), is also the receptor for iC3b. We studied the role of the cell adhesion molecules in the NK effect on targets with and without C3 fragments. We focused on the E/T interaction of opsonized cells in the presence of anti CD18 mAb. mAb directed to the CD11a molecule caused 0 to 30% inhibition of the lysis of both non-opsonized and opsonized cells whereas the mAb antibody directed to the CD11c molecule had no effect. Reagents reactive with the iC3b binding site of CD11b (alpha-chain of the CR3) molecule did not alter the lysis of non-opsonized targets whereas they abrogated the C3-mediated increment of the Nk effect on opsonized cells. Two mAb preparations, 60.3 and IB4, directed to the CD18 chain shared by the three cell adhesion molecules abrogated in a dose dependent way the lysis of both non-opsonized and opsonized targets. The 60.3 mAb inhibited the iC3b binding site of CR3 (despite its localization on the alpha chain) and in accordance it inhibited the binding of lymphocytes to the opsonized target also. The IB4 did not affect this site and in accordance it inhibited only partially the binding of effectors to the C3 fragment carrying Raji, nevertheless it inhibited their lysis. This result indicates that the iC3b-CR3 bridge is insufficient for triggering the lysis in absence of the contact through the adhesion molecules. PMID- 2565932 TI - Dendritic epidermal T cells: activation requirements and phenotypic characterization of proliferating cells. AB - Dendritic epidermal T cells (DETC) are CD45+, Thy-1+, CD5-, CD8-, CD4- murine lymphocytes that express surface-bound CD3 antigens associated with T cell receptor gamma/delta heterodimers. Using epidermal cells greatly enriched for DETC and depleted of Langerhans cells, we found that DETC have growth requirements quite different from those of accessory cell-depleted lymph node and splenic T cells. Although the latter cells strongly proliferate in response to phorbol myristate acetate (PMA) + ionomycin, DETC, when exposed to interleukin-1 (IL-1), interleukin-3 (IL-3), concanavalin A (ConA), PMA, and ionomycin used either alone or in combination, do not exhibit significant mitotic activity. Recombinant interleukin 2 (rIL-2), albeit ineffective by itself, leads to vigorous proliferation of DETC when used with either ConA or PMA + ionomycin + IL 1. In contrast, the combination of PMA and recombinant interleukin-4 (rIL-4), which triggers growth of lymph node T cells, does not induce proliferation of DETC. Although a portion of proliferating DETC expressed CD8 antigens, essentially none bore detectable amounts of surface-bound CD4 or CD5 antigens, or both. Continuing stimulation of primary DETC cultures with lectin/lymphokine-rich media results in the propagation of cells with the essential phenotypic features of resident DETC. PMID- 2565933 TI - Loss of heterozygosity at polymorphic chromosomal loci in patients with malignant melanoma. AB - One type of aberration of DNA found in human cancers is loss of heterozygosity at chromosomal loci. This loss suggests that inactivation of particular genes is involved in the genesis of cancer cells. We have analyzed tumor DNA and normal DNA from eight patients with malignant melanomas using 24 polymorphic DNA probes and detected loss of heterozygosity in three patients at five loci on four different chromosomes. These results indicate that the frequency of loss of heterozygosity at chromosomal loci is relatively high in malignant melanomas, but that the regions lost are not restricted to specific chromosomes. PMID- 2565931 TI - Effect of psoralens and ultraviolet radiation on murine dendritic epidermal cells. AB - Monofunctional psoralens produce less phototoxicity than bifunctional psoralens after ultraviolet A (UVA) irradiation. We investigated the effect of repetitive treatments with angelicin (isopsoralen), a monofunctional psoralen, plus UVA radiation (IPUVA) on the number and morphology of dendritic epidermal cells (dEC). This effect was compared with that of 8-methoxypsoralen plus UVA radiation (PUVA), UVA alone, and UVB radiation. C3H/HeN mice were treated topically with the drugs three times/wk for 4 consecutive wk; followed each time by 1 or 2.5 J/cm2 of UVA radiation. Other groups of mice were treated with the drugs alone, UVA alone, or 0.81 J/cm2 of UVB. Epidermal sheets were stained for ATPase, Ia, and Thy-1 markers. Mice treated with PUVA and UVB exhibited severe phototoxicity, whereas no overt phototoxicity was observed in mice treated with IPUVA, UVA alone, or the drugs alone. Early during the PUVA and UVA treatments the ATPase marker was lost from dEC, followed by loss of the Ia marker; the Ia marker was lost before the ATPase marker from dEC in animals treated with IPUVA. At the end of the treatment, however, nearly total depletion of ATPase+, Ia+, and Thy-1+ dEC was observed in mice treated with PUVA and IPUVA. UVB radiation caused rapid depletion of Thy-1+ dEC as well as ATPase+ and Ia+ cells. During treatments with IPUVA, PUVA, UVA, and UVB, the Langerhans cells became rounded and lost their dendrites. These changes were quantitated by image analysis. We conclude that alterations of cutaneous immune cells can occur in the absence of overt phototoxicity, and that monofunctional and bifunctional psoralens plus low dose of UVA radiation may have different effects on dEC markers. PMID- 2565934 TI - E.s.r. studies on the mechanism of hydroxyl radical-induced strand breakage of polyuridylic acid. AB - Reaction of photolytically produced .OH radicals with polyuridylic acid [poly(U)] in neutral solutions resulted in the electron spin resonance (e.s.r.) spectrum of the C(5)-OH-6-yl (= 6-yl) radical 1b of the nucleobase. At pH less than or equal to 4 the spectrum of the base radical had disappeared and instead the cyclic 2' oxo-3'-yl sugar radical 2a was observed. The assignment of the sugar radical was supported by model reactions with SO4-. as the radical inducing agent. Time resolved e.s.r. measurements showed that the rate of decay of the 6-yl base radical at neutral pH is virtually the same as that of the strand break (sb) formation. These results prove that: (i) in agreement with an earlier proposal the C(2') mechanism contributes to sb formation of poly(U), and (ii) the decay of the 6-yl radical is the rate-determining step in the reaction sequence leading to strand breakage. The change in the e.s.r. spectra at pH 4 is due to an increase in the rate of sb formation with increasing proton concentration. This effect is explained by generation of the radical cation, 4, from the 6-yl radical and/or by rearrangement of the 6-yl radical into the C(6)-OH-5-yl (= 5-yl) radical in proton-induced reactions and subsequent rapid H abstraction from the sugar moieties by species 4 and/or 5. PMID- 2565935 TI - Post-ultraviolet DNA synthesis in the absence of repair: role of the single strand DNA-binding protein. AB - Post-ultraviolet DNA synthesis kinetics were investigated in the Escherichia coli uvrA recA strain and its isogenic counterpart, overproducing single-strand DNA binding protein (SSB). It was demonstrated that large quantities of SSB enhance the capacity of the unmodified replisome to use the UV-damaged template for DNA synthesis. DNA thus synthesized is of low molecular weight, as shown by sedimentation in alkaline sucrose gradients. It is therefore suggested that SSB actively participates in the replisome translocation past dimers and/or the initiation of new DNA chains downstream of these lesions. PMID- 2565937 TI - DNA damage and repair in rodent and human cells after exposure to JANUS fission spectrum neutrons: a minor fraction of single-strand breaks as revealed by alkaline elution is refractory to repair. AB - We have examined the induction and repair of breaks induced in the DNA of Chinese hamster V79 and human P3 epithelial teratocarcinoma cells by JANUS fission spectrum neutrons (mean energy 0.85 MeV) and 60Co gamma radiation in the biological dose range, using alkaline filter elution methods. Fission-spectrum neutrons induce fewer immediate single-strand breaks (ssb) per gray of absorbed dose than do gamma rays, as measured by alkaline elution methods. Previous survival measurements have indicated incomplete recovery after neutron exposures. The present data demonstrate that whereas most ssb caused by exposure to fission spectrum neutrons can be rapidly repaired by both cell lines, a small but statistically significant fraction of the ssb induced by exposure to 6 Gy of neutrons is refractory to repair. In contrast, all measurable ssb induced by 3 Gy gamma rays are rapidly repaired. PMID- 2565938 TI - Inhibition of X-ray-induced DNA strand break repair in polyamine-depleted HeLa cells. AB - Treatment of HeLa cells with the polyamine biosynthesis inhibitors, alpha difluoromethylornithine (DFMO) or methylglyoxal bis(guanylhydrazone) (MGBG), results in, depending on the conditions, partial or complete depletion of the cellular polyamines: putrescine, spermidine and spermine. In this compromised state cells exhibited a distinct deficiency in repair of X-ray-induced DNA strand breaks. The half-time for return of normal DNA sedimentation following 1.6 Gy was 9.5 min for untreated control cells and 22, 32 and 50 min for cells treated with MGBG, DFMO + MGBG and DFMO, respectively. Normal repair kinetics were restored to these cells upon a short incubation in media containing all three polyamines. The rapid early phase of repair following higher X-ray doses (16 Gy) was also delayed in polyamine-depleted cells but later repair occurring 1-4 h post-irradiation, representing chromatin reconstitution, was apparently normal. PMID- 2565936 TI - Detection of DNA damage in cells exposed to ionizing radiation by use of anti single-stranded DNA monoclonal antibody. AB - An immunochemical method has been developed for quantitative detection of DNA damage in mammalian cells. The method is based on the binding of a monoclonal antibody to single-stranded DNA. The clone producing this antibody (D1B) was obtained as a by-product from fusion of mouse myeloma cells with spleen cells isolated from a mouse immunized with chemically modified DNA. The technique is based upon the determination of the percentage single-strandedness resulting from the time-dependent partial unwinding of cellular DNA under alkaline conditions. Single- and double-strand DNA breaks, or lesions converted into such breaks in alkaline medium, form initiation points for the unwinding. The extent of unwinding from these points under defined conditions is a measure of the number of such sites. The method is rapid, does not require radioactive labelling of DNA or physical separation of single-from double-stranded molecules, is sufficiently sensitive to detect damage induced by 1 Gy of ionizing radiation and needs only small numbers of cells. The usefulness of the technique was demonstrated in a study of the induction of DNA damage and its repair in cultured Chinese hamster cells and in human white blood cells after exposure to 60Co-gamma-rays, and in white blood cells and bone marrow cells of X-irradiated mice. A dose-related DNA unwinding was observed and repair of DNA lesions was observed up to 60 min after irradiation. PMID- 2565939 TI - raf involvement in the simultaneous genetic transfer of the radioresistant and transforming phenotypes. AB - We examined a human Alu+ mouse tertiary transformant derived from a noncancerous skin fibroblast cell line which exhibits the unique characteristic of being resistant to the killing effects of ionizing radiation. This transformed cell line was found to contain activated human c-raf-1, and demonstrated an increased level of radioresistance indicating the simultaneous transfer of both the transforming and radiation-resistant phenotypes. We have also found a relationship between the presence of activated oncogenes, specifically those with serine/threonine kinase activity and the radioresistant phenotype. PMID- 2565940 TI - Radiation-induced diploid spermatids in mice. AB - Diploid elongated spermatids of mice were enriched by flow cytometry and cell sorting using a new type of sorter (Partec). The sorted abnormal spermatids were identified morphologically and by nuclear area integration. The radiation-induced increase in the frequency of diploid elongated spermatids was monitored with time following acute X-ray exposure of mice. Dose-response curves for acute 60Co-gamma and 14 MeV neutron irradiations yielded an RBE value of 4.3 for the doubling of the control level. PMID- 2565941 TI - The independent action of radiation and cisplatin on the survival or recovery of human normal or tumour cells in vitro or in vivo. AB - Recovery from radiation- or cisplatin-induced lethal damage has been studied in euoxic normal human foetal lung fibroblasts (HFL cells) that remain viable for at least 20 days in plateau-phase culture. After a 1 hour treatment with cisplatin the half-time of recovery was about 2 days. By contrast recovery after radiation was more rapid with half-times of approximately 10 h. There was no further measurable recovery after 2 days. With either agent the recovery ratios (RR) were dose-dependent but recovery (following treatment with equitoxic doses of the two agents) was appreciably greater after cisplatin (RR approximately 123 after 40 microM for 1 h) than after radiation (RR approximately 15 after 900 cGy). When radiation (900 cGy) was combined with cisplatin (40 microM for 1 h) the cell survival, measured at 5 days or later times after the treatments, was not significantly less than that predicted by the additive, independent effects of both agents (calculated as the product of their respective effects on cell survival) irrespective of whether cisplatin was given 1 h before, during or for 1 h immediately after radiation. In euoxic, exponentially growing HFL or HeLa cells there was no evidence that combinations of cisplatin and radiation gave more than additive toxic effects in the protocols tested. The combined effects of cisplatin or carboplatin and whole-body irradiation given 45 min later, on human melanoma cells (assessed by their colony-forming abilities in vitro) growing in thymectomised mice, were essentially the same as that predicted by the additive, independent effects of the two agents. PMID- 2565942 TI - The effect of fetal irradiation on the growth of postnatally xenotransplanted tumour cells. AB - Exposure of the mouse fetus (NMRI-strain) to 1.0 Gy X-irradiation has a marked effect on postnatally xenotransplanted glioma cells. In comparison to non irradiated animals, irradiation on gestation day 14 resulted in: (a) a significantly higher rate of animals which failed to develop visible tumours growing from the inoculum; (b) a significant inhibition of the growth rate of solid gliomas; (c) a pronounced granulocytic and mast cell infiltration, and tissue necrosis, in the invading gliomas. The results suggest that irradiation in prenatal life exerts an amplifying effect on the antitumour response in postnatal life. PMID- 2565943 TI - Behaviour of spherical and irregular (U,Pu)O2 particles after inhalation or intratracheal instillation in rat lung and during in vitro culture with bovine alveolar macrophages. AB - Spherical and irregularly shaped mixed (U,Pu) oxide particles were administered to rats by inhalation and by intratracheal instillation. The lung retention of the particles was independent of particle shape and of the route of administration. Only a small percentage of the administered radioactivity was found in other organs. Detailed electron microscopic studies showed particles within membrane-enclosed vacuoles as well as lying free in the cytoplasm. PMID- 2565944 TI - Changes in oligosaccharides of human blood cells measured by tritium- and FITC labeled lectins after in vitro- and in vivo-irradiation--lectin binding as a 'biological dosemeter'? AB - The lectin-binding system has been described previously as a biological dosimeter, by revealing induced changes in oligosaccharides of the cell membrane. The measurements were performed by binding [3H]concanavalin A to blood cells. Our results on human blood cells irradiated in vitro with doses in the range 0.5-5 Gy indicate great difficulties in using radioactive labeled Con-A for an accurate quantitative analysis of radiation effects on cell membranes. It appears nearly impossible to differentiate between only a few damaged cells and the remaining undamaged cells. Using fluorescein-labeled Con-A and wheat germ agglutinin, single-cell measurements of fluorescence intensity by flow cytometry revealed enhanced lectin-binding to platelets, lymphocytes and monocytes in the dose range 0.5-5 Gy after in vitro irradiation. But even by this method it was impossible to discriminate irradiations in either partial or whole-body irradiated patients. There were no significant or reproducible changes in the binding capacities of the blood-cell membranes of these patients. Therefore, the suitability of lectin binding as a 'biological indicator' for irradiation could not be confirmed. PMID- 2565945 TI - Chromosomal aberrations in cynomolgus peripheral lymphocytes during and after fractionated whole-body gamma-irradiation. AB - Cynomolgus monkeys (Macaca fascicularis) were exposed to fractionated whole-body gamma-irradiation at high and low dose rates for 4 or 5 weeks. The time dependence of chromosomal aberrations was studied in relation to the number of lymphocytes during irradiation and after exposure for periods of up to about 600 days for chromosomal aberrations and 200 days for lymphocyte counts. Additivity of the daily effects on the number of chromosomal aberrations was observed during the exposures. Immediately after the end of the exposures the number of chromosomal aberrations decreased to reach low values. The disappearance of chromosomal aberrations seemed to be related to recovery of the lymphocyte counts. The data presented here emphasize the different kinetic patterns of chromosomal aberrations after fractionated and acute irradiation. PMID- 2565946 TI - Effect of gentamicin on the carbohydrates of the vestibular end organs: an investigation by the use of FITC-lectins. AB - The effect of gentamicin on the glycoconjugates in the vestibular end organs is demonstrated using FITC-lectins. Five milligrams of gentamicin dissolved in 0.1 ml. saline was injected in a single dose into the middle ear of adult guinea pigs. Seven days after the injection, the fluorescent reactivity of Wheat germ agglutinin (WGA), Abrus precatorius agglutinin (APA), Concanavaline A (Con A), and Latyrus odoratus agglutinin (LOA) was decreased in the apical epithelial cell surface as well as in the gelatinous layer of the otolithic membrane of the maculae. In the cupula of the crista ampullaris, the reactivity for WGA and Con A was reduced. The otoconia, however, showed no detectable changes. This indicates that gentamicin may induce an altered carbohydrate metabolism resulting in a decrease of N-acetyl-glucosamine, mannose, galactose and glucose in the glycocalyx lining the epithelial cells as well as in the gelatinous layer of the otolithic membrane or cupula in the vestibular end organs. PMID- 2565947 TI - Plasma concentrations of cholecystokinin octapeptide and food intake in male rats treated with cholecystokinin octapeptide. AB - Intraperitoneal injection of 5 micrograms cholecystokinin octapeptide (CCK-8) into male rats deprived of food for 48 h produced a transient (less than 15 min) increase in plasma levels of CCK-8 but suppressed food intake for an extended period (45 min). Plasma concentrations of CCK-8 after i.p. injection of CCK-8 were raised to levels which were fairly comparable to those after feeding. Intracerebroventricular (i.c.v.) injection of the CCK antagonist proglumide (100 micrograms) reversed the effect of CCK-8 on food intake, while i.p. injection of proglumide (100 micrograms) did not have this effect. Feeding increased the plasma concentrations of somatostatin and gastrin but not of oxytocin, and somatostatin and oxytocin but not gastrin were released in response to i.p. injection of CCK-8. However, neither somatostatin nor oxytocin affected food intake, and their release in response to CCK-8 was unaffected by i.c.v. injection of proglumide. These results support the suggestion that CCK-8 is a physiological 'satiety' peptide, which can affect food intake in rats by mechanisms involving both peripheral and central CCK receptors. PMID- 2565948 TI - Transient adhesion of neutrophils to endothelium. AB - Fluorescently labeled polymorphonuclear leukocytes (PMN) were used to measure adhesion to human umbilical vein endothelial cells (EC) cultured in vitro. Stimulation of PMN with phorbol dibutyrate (PDB), TNF, or C5a caused an increase in adhesion followed by a return to prestimulation levels of adhesion of longer times of incubation. Maximal adhesion of PMN to EC occurred rapidly in response to C5a (5 min) and more slowly with TNF or PDB (15 min). PMN stimulated to adhere with C5a detached from EC by 15 min. PMN from CD11/CD18-deficient patients and PMN incubated with anti-CD18 mAbs failed to bind to EC despite maximal stimulation. Anti-CD11a/CD18 and anti-CD11b/CD18 each partially inhibited adhesion, and a combination of these two reagents completely blocked adhesion. The adhesion we measured was therefore completely dependent on CD11/CD18, and CD11a/CD18 and CD11b/CD18 each contributed to adhesion. Stimuli that enhanced adhesion of PMN to EC also enhanced expression of CD11b/CD18 on the cell surface, but the time course of expression correlated poorly with changes in adhesivity. To determine if changes in the expression of CD11b/CD18 are necessary for the changes in adhesivity, we used enucleate cytoplasts that did not increase expression of CD11b/CD18. Cytoplasts showed a normal rise and fall in adhesivity in response to PDB. We conclude that the transient adhesion of stimulated PMN to naive EC is regulated by changes in the nature of existing CD11/CD18 molecules on the PMN surface. Changes in expression of CD11b/CD18 may contribute to enhancement of adhesivity, but a definite role for this phenomenon has yet to be established. PMID- 2565950 TI - Multi-drug theophylline overdose. PMID- 2565951 TI - [Genetic counseling on a molecular basis for Huntington's chorea]. AB - The gene responsible for Huntington Disease has been mapped at the end of the short arm of chromosome 4. Different polymorphic probes from the immediate vicinity of the gene are available and provide accurate tools for the identification of carriers of the gene. The presymptomatic testing of adults or the prenatal testing (exclusion testing) of the unborn child must be considered with great caution, it requires a multidisciplinary approach and will benefit from intensive collaboration with the lay organizations. Presymptomatic testing will presumably find a much wider application when it can be coupled to an effective preventive treatment. PMID- 2565949 TI - Direct observation of the gene organization of the complement C4 and 21 hydroxylase loci by pulsed field gel electrophoresis. AB - Pulsed field gel electrophoresis and enzymes that cut genomic DNA infrequently have been used to define large RFLPs at the human C4 loci. With the enzymes BssH II or Sac II, and C4 or 21-hydroxylase DNA probes, it has been possible to observe directly the number of C4 genes present on a haplotype, and also whether the C4 genes are long (6-7-kb intron present) or short (6-7-kb intron absent). Haplotypes that have either two long C4 genes or one long and one short C4 gene generate BssH II fragments of approximately 115 or approximately 105 kb, respectively. Haplotypes that have either a single long or a single short C4 gene generate BssH II fragments of approximately 80 or approximately 70 kb, respectively. This technique has been used to analyze the DNA isolated from PBMC and allows the complete definition of the C4 gene organization of an individual without the need for family studies. PMID- 2565952 TI - [Contribution of molecular biology to the prevention of cystic fibrosis. Experience in Lyon]. AB - Enzymatic prenatal diagnosis of cystic fibrosis was performed in 113 amniotic fluids and DNA polymorphism was studied in 104 families, including 28 cases with prenatal material analysis. According to the results, the enzymatic diagnosis should be cautiously interpreted when the risk is less than 1/4. In these situations DNA analysis in the parents is very helpful to assess the reliability of enzymatic diagnosis. PMID- 2565953 TI - [Myotonic dystrophy of Steinert]. AB - The gene for myotonic dystrophy maps to 19q13.2----19q13.3. The closest proximal marker is the gene for creatine kinase CKMM at a recombination faction of 0-2%. Prenatal diagnosis will be performed with a minimal risk when a distal closer to the gene is available. PMID- 2565954 TI - Prognostic factors and staging classification of patients with epidemic Kaposi's sarcoma. AB - Two hundred twelve patients with acquired immune deficiency syndrome (AIDS) related Kaposi's sarcoma (KS) were followed prospectively. Univariate and multivariate analyses were performed to determine significant predictors of survival and development of opportunistic infection (OI) from the time of diagnosis of KS. Clinical variables analyzed were age at onset, presence of systemic symptoms, prior or coexistent OI, development of OI greater than 3 months following KS diagnosis, and extent of disease. Laboratory variables analyzed were absolute number of peripheral T-helper lymphocytes (T4), helper/suppressor ratio (T4/T8), serum beta-2-microglobulin, and serum acid labile alfa interferon. Three independent variables were predictive of shorter survival: (1) prior or coexistent OI (P = .02), (2) presence of systemic symptoms (P = .001), and (3) absolute T4 count less than 300 cells/microL (P = .002). Based on survival, patients with AIDS-related KS can be divided into four groups: (1) those with no prior or coexistent OI, no systemic symptoms, T4 greater than or equal to 300 cells/microL (median survival, 31 months): (2) those with no prior or coexistent OI, no systemic symptoms, and T4 less than 300 cells/microL (median survival, 20 months); (3) those with no prior or coexistent OI and presence of systemic symptoms (median survival, 15 months); and (4) those with prior or coexistent OI (median survival, 7 months). PMID- 2565955 TI - Development of dehydroemetine nanoparticles for the treatment of visceral leishmaniasis. AB - The preparation and physico-chemical characterization of lyophilized polyisohexylcyanoacrylate nanoparticles loaded with dehydroemetine (DHE) for the treatment of visceral leishmaniasis disease is described. The resulting formulation was found to efficiently absorb DHE and gave very reproducible preparations with regard to the size and drug adsorption rate. Stability has been confirmed for at least 24 months. The acute toxicity of DHE was reduced in intravenous administration by its association with nanoparticles. Data concerning tissue distribution in mice showed that DHE nanoparticles were rapidly cleared from the blood stream and that they mainly concentrated in the reticuloendothelial system. Furthermore, DHE linkage to the carrier reduced the cardiac concentrations of the drug. PMID- 2565956 TI - Thermal characterization of drug/polymer and excipient/polymer interactions in some film coating formulation. AB - Some probable consequences of the dissolution/migration of a major solid dosage component in or into an applied film coating during or after a film coating operation have been investigated using free films of hydroxypropyl methylcellulose (HPMC) and polyvinyl alcohol (PVA) incorporating small amounts of either lactose (a diluent) or ephedrine hydrochloride (a drug). Intrinsic features of the films such as softening, glass transition, crystallinity and melting were examined by differential scanning calorimetry and thermomechanical analysis. Generally, the results indicate that ephedrine hydrochloride exhibited plasticizing activity in both HPMC and PVA films. On the other hand, incorporation of lactose produced an opposite effect (stiffening) in PVA films as demonstrated by increased glass transition temperature and crystallinity. On the basis of these findings, it was proposed that the undesired presence of a component of a solid dosage core in the applied film coating could significantly alter its end-use properties such as diffusivity and the incidence of film coating defects. It was also shown that the application of the relationship of Moelter & Schweizer (1949) in the evaluation of the plasticizer efficiency of non homologous additives could pose problems of interpretation. PMID- 2565957 TI - Chemical reactivity of aluminium-based pharmaceutical compounds used as phosphate binders. AB - Several aluminium-containing substances, including antacids used as phosphate binders in treating renal failure, have been analysed in-vitro under different pH conditions for the release of Al3+ ions and for binding of phosphate. Control experiments on different forms of pure aluminium hydroxide validated the methods. At pH 2 it was the most amorphous forms which released Al3+ most rapidly. These aluminium ions, available for absorption by the patient, were released from all antacids tested, but no firm phosphate-binding was detected while the pH remained at 2. Phosphate was bound at pH 8, by adsorption onto the surface of aluminium hydroxide. No significant amounts of free Al3+ exist in solution at pH 8, since at that pH aluminium hydroxide is precipitated. The most amorphous forms of this solid were the most efficient phosphate-binders. Alumino-silicate salts require prior exposure to acid to produce free Al3+ before they can act as phosphate binders, whereas amorphous aluminium hydroxide acts as an efficient phosphate binder without prior exposure to acid. Chemical principles are employed to show why aluminium release and phosphate-binding are separate and independent processes. Methods are proposed for maximizing the activity of phosphate- binders in-vivo, while minimising aluminium release. PMID- 2565958 TI - Rate and extent of absorption of clonidine from a transdermal therapeutic system. AB - The in-vivo performance of a clonidine transdermal therapeutic system (TTS 3.5 cm2, 2.5 mg) was assessed in 12 healthy normal volunteers. Particular attention was paid to the rate and extent of absorption of clonidine from the TTS dosage form by reference to a 2 h i.v. infusion of clonidine. The absolute bioavailability of clonidine from the TTS dosage form was found to be approximately 60% with clonidine being released from the TTS at a relatively reproducible and consistent rate of 4.32 micrograms h-1 over a 7-day period. PMID- 2565959 TI - The interaction of papain with polycations. AB - An investigation has been made of the interactions of the enzyme papain with the polycations protamine, polybrene, poly(L-lysine), spermine, spermidine and the neutral polymer polyvinylpyrrolidone (PVP). At low concentrations, each behaves as an inhibitor of the enzyme. As their concentrations increased above a certain level, the activity of the systems increased, and their inhibition of the enzyme appeared to be less pronounced. When acting by themselves in the presence of the substrate haemoglobin, each of the polycations was a weak proteolytic catalyst with a ranking of catalytic effectiveness of protamine greater than polybrene greater than poly(L-lysine) greater than polyvinyl-pyrrolidone greater than spermidine greater than spermine. This effect could explain the anomalous inhibition of papain by these polycations. The interaction of papain with dansyl protamine (DNSP) and the extent of complex formation were studied using a fluorescence polarization technique and the results showed that there was a strong interaction occurred. The strength of binding was assessed by determination of the critical electrolyte concentration (0.2 M, NaNO3). The stoichiometry of the DNSP-papain complex was found to be 63 base moles of DNSP to one mole of papain. PMID- 2565960 TI - Unequal disposition of enantiomers of the organic cation oxyphenonium in the rat isolated perfused liver. AB - This paper describes the results of pharmacokinetic experiments in the rat isolated perfused liver with enantiomers of oxyphenonium. The study was performed with the [14C]methyl labelled compounds. In this preparation both metabolism and biliary excretion were significantly different for the (+)- and the (-)-isomer. Hepatic uptake rate was similar, but total biliary excretion (including metabolites) of the (-)-isomer was only 55% compared with the excretion of the (+)-isomer. In line with these data, after 2 h only 30% of the dose of the (+) isomer and over 50% of the dose of the (-)-isomer was still found in the liver, predominantly in the form of metabolites. The metabolic profile was investigated using ion pair TLC. At least two metabolites were detected in bile for both enantiomers. However, unchanged (-)-oxyphenonium persisted for longer in bile, indicating either a more rapid canalicular transport of the (+)-isomer and/or a more rapid metabolism of (+)-oxyphenonium to cholephilic metabolites. PMID- 2565961 TI - Comparative studies of the effects of RS-8359 and safrazine on monoamine oxidase in-vitro and in-vivo in mouse brain. AB - The effect of RS-8359, pyrimidine on monoamine oxidase (MAO) has been compared with a hydrazinic MAO inhibitor, safrazine (beta-piperonylisopropylhydrazine hydrochloride,) which is a MAO inhibitor used clinically. In-vitro radiochemical determination of MAO activity showed that the IC50 of RS-8359 was 0.52 microM for the deamination of 5-hydroxytryptamine (5-HT) in the mouse brain mitochondrial preparation, while beta-phenylethylamine (PEA) deamination was inhibited by only 20% at 100 microM of the drug. 5-HT deamination in the brain homogenate prepared from mice killed 60 min after administration of RS-8359 was inhibited significantly by 14 and 48%, at 30 and 100 mg kg-1 (p.o.), respectively, while deamination of PEA was little affected at the same doses. On the other hand, safrazine strongly inhibited both 5-HT and PEA deaminations, but showed no selectivity toward the substrate used. The extent of MAO inhibition by RS-8359, measured fluorometrically with kynuramine as a substrate in the brain homogenate, was independent of preincubation up to 80 min. In contrast, the inhibitory potency of safrazine was strengthened by preincubation in a time-dependent manner. Oral administration of RS-8359 (3-30 mg kg-1) caused a dose-dependent increase in endogenous monoamines in mouse brain, which disappeared a few hours after its administration. Increase in monoamine content caused by safrazine lasted for at least 24 h. These results indicate that RS-8359 is a reversible and specific inhibitor of MAO-A, while safrazine is an irreversible and non-specific MAO inhibitor, in-vivo and in-vitro in mouse brain. PMID- 2565962 TI - In-vivo effects of (E)-2-(3',4'-dimethoxyphenyl)-3-fluoroallylamine (MDL 72145) on amine oxidase activities in the rat. Selective inhibition of semicarbazide sensitive amine oxidase in vascular and brown adipose tissues. AB - One hour after MDL 72145 ((E)-2-(3',4'-dimethoxyphenyl)-3-fluoroallylamine) (2.5 mg kg-1) was given by intraperitoneal injection, the semicarbazide-sensitive amine oxidase (SSAO) activity of rat aorta and brown adipose tissue measured in vitro was reduced by more than 95% of its control value, whereas the monoamine oxidase (MAO-A) activity remained virtually unaffected. The action of this drug on amine oxidases in the liver at this dose was less selective. The in-vitro effect of MDL 72145 on the soluble enzyme diamine oxidase from rat intestine was 100 fold less potent than that of semicarbazide but about equipotent with semicarbazide on sheep plasma amine oxidase. Overall MDL 72145 was selectively more active against membrane bound SSAO enzymes that deaminate primary monoamines. Although MDL 72145 does inhibit MAO-B activity these results suggest that this compound may be used to study the effect of selective inhibition of SSAO activity on the pharmacological responses of appropriate preparations in vitro. PMID- 2565963 TI - The effect of diphenylamine-2-carboxylate on C1- channel conductance and on excitability characteristics of rat skeletal muscle. AB - The effect of diphenylamine-2-carboxylate (DPC), a blocker of the C1- conductive pathway in C1- transporting epithelia, has been evaluated in-vitro on the electrophysiological variables of rat extensor digitorum longus muscle fibres. DPC (5-240 microM) caused a dose-related increase of membrane resistance which was attributed entirely to a fall in C1- channel conductance (IC50, 120 microM), since potassium conductance was not affected by the treatment. DPC also modified fibre excitability. A significant dose-dependent increase was observed in the latency of the action potential and in the excitability of the membrane. DPC was less potent on striated fibres than anthracene-9-carboxylic acid, another specific blocker of C1- channel conductance. Moreover DPC was less potent on skeletal muscle than on C1- transporting epithelia. Morphological differences in the C1- channels or of the drug binding sites may account for the differences between tissues. PMID- 2565964 TI - Mediators of the plasma extravasation response to silver nitrate in the rat skin, subplantar region and ankle joint. AB - Plasma extravasation responses to silver nitrate (AgNO3), histamine, 5 hydroxytryptamine (5-HT), bradykinin and prostaglandin E1 (PGE1) in the abdominal skin, hindpaw ankle joint and subplantar region of rats have been investigated using the Evans blue dye leakage technique. All substances tested produced plasma extravasation and combination of low doses (5 x 10(-10) mol) of either histamine or bradykinin with PGE1 (5 x 10(-10) mol) exhibited potentiation of responses of all regions. Responses to AgNO3 (1 x 10(-6) mol) were significantly reduced by the H1 receptor antagonist, mepyramine, only in the abdominal skin, but the H2 receptor antagonist metiamide reduced the responses at subplantar and ankle joint regions. Indomethacin significantly reduced the AgNO3 responses at the ankle joint only, but aprotinin reduced it at the other two regions. In rats pretreated with a combination of all antagonists the residual plasma extravasation response to AgNO3 was very small, indicating that the response could be almost totally accounted for by the combined actions of mast cell amines, kinins and prostanoids. The finding that prostanoids played a major role in the plasma extravasation response of the rat ankle joint to AgNO3 indicated that this model would be useful for the screening of non-steroidal anti-inflammatory drugs. PMID- 2565965 TI - The effect of some tricyclic antidepressants on the inhibition of mouse brain monoamine oxidase in-vivo by phenelzine. AB - Four tricyclic antidepressants, amitriptyline, imipramine, desipramine and iprindole have been shown to partially protect mouse brain monoamine oxidase in vivo from the irreversible enzyme inhibition produced by subsequent injection of phenelzine. Levels of protection were similar when the enzyme was assayed with selective substrates (5-hydroxytryptamine and phenethylamine) for both the A and B forms of the enzyme. Although other explanations cannot at this stage be ruled out, these observations are consistent with the tricyclic antidepressants acting as reversible inhibitors of brain monoamine oxidase in-vivo. PMID- 2565966 TI - Hypothermia induces supersensitivity of mouse vas deferens to adrenergic agonists: evidence for postjunctional alpha 2-adrenoceptors. AB - Hypothermia-induced potentiation of alpha-adrenoceptor-mediated responses of the mouse vas deferens to noradrenaline is mainly caused by inhibition of the sites of loss. However, even after blockade of the sites of loss for noradrenaline or when using methoxamine (which is not a substrate for uptake or metabolism) hypothermia still causes a significant increase in responsiveness. This remaining supersensitivity was shown to be an increased receptor affinity. Furthermore, hypothermia revealed contractile responses to low concentrations of UK 14304 (an alpha 2-agonist). PMID- 2565967 TI - The effects of B-HT 920 and St 91 on venous haemodynamics in cats. AB - The present study reports the effects of 2-amino-6-allyl-5,6,7,8-tetrahydro-4H thiazolo-[4,5-d]azepin dihydrochloride (B-HT 920) and 2-(2,6-diethylphenylimino) 2-imidazolidine hydrochloride (St 91) in autoperfused cat hindquarters since we have previously shown that clonidine lowered blood pressure, heart rate and vena cava blood flow (VCBF) but not hindquarters perfusion pressure, indicating a selective venodilator action of this drug (Bentley et al 1986). It was found that intravenous (i.v.) and intracisternal (i.c.m.) administration of B-HT 920 caused essentially identical effects to those of clonidine. St 91, given i.c.m. lowered blood pressure and VCBF but not perfusion pressure, while i.v. St 91 had little effect on these variables. Thus, parallel changes in blood pressure and VCBF occurred using both drugs, suggesting that these centrally-acting clonidine-like drugs also caused selective venodilatation. PMID- 2565968 TI - QSAR study on species differences in microsomal N-oxygenation of N,N dimethylalkylamines. AB - The metabolic N-oxygenation of nine long chain N,N-dimethylalkylamines and tri-n butylamine has been studied using hepatic microsomal homogenates from mice, dogs and guinea-pigs. The relative oxidizability of amines (ROA) was correlated with structure, lipophilicity and nucleophilicity parameters of substrates and corresponding amine oxides formed in this biological reaction. The highest conversion of amines to N-oxides was found with male guinea-pig, followed by dog (male, female) and the lowest amount of amine oxides has been produced with male mice microsomal homogenates. The analyses were carried out by GLC and the results quantified using QSAR methodology. ROA is parabolically dependent upon structure and physicochemical properties of the substrates and products which was proved by the high statistical significance of the regression equations. The biological N oxygenation of these amines is controlled by lipophilicity, stereochemistry and electronic effects. PMID- 2565969 TI - Doxorubicin is a potent inhibitor of interleukin 1 induced cartilage proteoglycan resorption in-vitro. AB - Since interleukin 1 (IL-1) induces the transcriptional synthesis of enzymes responsible for cartilage resorption it was decided to examine the effects of the antitumour drug, doxorubicin, a DNA transcriptional inhibitor, on alpha IL-1 induced cartilage--resorption in-vitro. Doxorubicin inhibited the resorption in a concentration-dependent fashion, an effect which was shown to be reversible. Fine structure of the chondrocytes was preserved by the doxorubicin treatment with IL 1 in contrast to the extensive cellular destruction evident in cartilage treated with IL-1 alone. [14C]doxorubicin was bound to cartilage proteoglycans, and this effect was promoted by treatment of the cartilage with IL-1. This binding of the drug may prevent access of the proteoglycans to destructive enzymes during the resorptive process induced by IL-1. PMID- 2565970 TI - Difference spectrophotometric assay of nitrazepam in tablet formulations. AB - A difference spectrophotometric procedure is described for the assay of nitrazepam in tablet formulations. The method is based on the measurement of absorbance at 282 nm of a solution of the tablet extract in 0.1 M hydrochloric acid (pH 1) relative to that of an equimolar solution in 0.1 M sodium hydroxide (pH 13). The method is precise and selective for nitrazepam in the presence of the tablet excipients and 2-amino-5-nitrobenzophenone, the principal hydrolysis product of nitrazepam. The absence of a constant isosbestic point in the difference spectrum of nitrazepam during hydrolysis in alkaline solution indicates the presence of a previously unrecognized intermediate hydrolysis product. PMID- 2565971 TI - Effects of chronic nicotine pretreatment on (+)-amphetamine and nicotine-induced synthesis and release of [3H]dopamine from [3H]tyrosine in rat nucleus accumbens. AB - The effects of chronic (14 day) administration of nicotine (1.5 mg kg-1 day-1) on the rat nucleus accumbens have been examined. Pretreatment of animals with nicotine increased the endogenous level of dopamine. The ability of (+) amphetamine to stimulate formation and release of [3H]dopamine from [3H]tyrosine was greatly potentiated in tissue slices from the nucleus accumbens of rats pretreated with nicotine. Furthermore, nicotine was effective in stimulating the formation and release of from [3H]dopamine from [3H]tyrosine in tissue slices from chronic nicotine-treated animals. PMID- 2565972 TI - Effects of tapioca obtained from cassava (Manihot utilissima) on the disintegration and dissolution rates of paracetamol tablets. AB - The properties of tapioca obtained from cassava (Manihot utilissima) have been evaluated. Its binding effect in tablets of paracetamol on the disintegration and dissolution rates was compared with tablets prepared with polyvinylpyrrolidone and gelatin. The nature and amount of the binders were found to alter the disintegration and dissolution rates of the tablets by reducing their wettability as measured by the adhesion tension of water. A linear relationship has been found to exist between the adhesion of water on the tablets and their disintegration and dissolution rates. PMID- 2565973 TI - Muscarinic receptor subtypes involved in bethanechol-induced water intake in the rat. PMID- 2565975 TI - Prodrugs of peptides. IV: Bioreversible derivatization of the pyroglutamyl group by N-acylation and N-aminomethylation to effect protection against pyroglutamyl aminopeptidase. AB - Various N-acyl derivatives and N-Mannich bases of the model compound L pyroglutamyl benzylamide were synthesized to assess their suitability as prodrug forms for the N-terminal pyroglutamyl residue occurring in several peptides, with the aim of improving peptide delivery characteristics. Whereas pyroglutamyl benzylamide was rapidly hydrolyzed by pyroglutamyl aminopeptidase, the N-acyl derivatives and N-Mannich bases (N-aminomethyl derivatives) were totally resistant to cleavage by the enzyme. On the other hand, these derivatives are readily bioreversible, the conversion to the parent pyroglutamyl amide taking place either by spontaneous hydrolysis at physiological pH, as demonstrated for the N-Mannich bases, or by plasma enzymes, as shown for the N-acyl derivatives. The results suggest that by appropriate N-acylation or N-aminomethylation it may be feasible to protect pyroglutamyl-containing peptides against cleavage by pyroglutamyl aminopeptidase and to obtain a release of the parent peptide in the organism, hence improving the delivery characteristics of such peptides. PMID- 2565974 TI - Nuclear magnetic resonance spectra and mosquito repellent properties of some oxazolidine derivatives of 5-methyl-4-hexenal. AB - Seven novel oxazolidines from 5-methyl-4-hexenal were synthesized, characterized, and evaluated in a cloth test system as mosquito repellents. These heterocycles were a homologous series of 3-alkanoyl-2-(4'-methyl-3'-pentenyl)oxazolidines, with substituents at N-3 ranging from acetyl to n-nonanoyl groups. Carbon-13 NMR spectroscopy at 100.6 MHz revealed that these molecules existed in deuterochloroform solution as unequal mixtures of cis (syn)- and trans (anti) rotational isomers. At 25 degrees C, this isomerism at the amide group resulted in two detectable 13C signals for all assigned carbon atoms, except for the methyl carbons of the isopentenyl side chain. Differences in 13C chemical shifts for the methyl group at N-3 were observed up to the n-butanoyl homologue. The 13C signal assignments were aided by the distortionless enhancement by polarization transfer (DEPT) spectral editing technique. Two-dimensional carbon-hydrogen chemical shift correlation experiments aided in partially interpreting the complex proton spectra of these molecules. Repellent activity of the acetyl, trideuteroacetyl, and propionyl derivatives was observed in bioassays with three mosquito species, but the long-chain homologues were practically ineffective as mosquito repellents. PMID- 2565976 TI - Immobilized sulfatase:beta-glucuronidase enzymes for the qualitative and quantitative analysis of drug conjugates. AB - The enzymes sulfatase and beta-glucuronidase from Helix pomatia were simultaneously immobilized on aminopropyl control pore glass. Once immobilized, these enzymes retained activity under varied conditions of pH, organic solvent, and temperature. To hydrolyze the sulfate and glucuronide conjugates of xenobiotics, the immobilized enzymes were either added directly to incubation mixtures for qualitative in vitro studies or packed in a short stainless steel column and placed in an HPLC system for quantitative studies. By incorporating specific inhibitors (D-saccharic acid-1,4-lactone to inhibit beta-glucuronidase or phosphate ions to inhibit sulfatase) into the incubation mixture or into the HPLC mobile phases, selective hydrolysis of either sulfate or glucuronide conjugates was achieved. Upon removal of the inhibitors from the incubation mixtures or from the mobile phases, original enzyme activity was restored. The utility of immobilized enzymes was demonstrated for quantitative analysis of sulfate and glucuronide conjugates of fenoldopam, where the liberation of the catechol aglycone moiety was necessary for electrochemical detection. PMID- 2565977 TI - Identification of secretin diastereoisomers produced during synthesis. AB - The byproducts P-1 and P-2, which were produced during the synthesis of porcine secretin, were isolated in pure form from the crude secretin by HPLC. These were identified by a combination of amino acid analysis, enzymatic digestion, and isocratic or linear gradient reversed-phase (RP)-HPLC. The amino acid compositions of P1 and P2, determined by amino acid analysis after acid hydrolysis, were found to be the same as those of porcine secretin without distinction between L-and D-amino acids. But, HPLC of their digestive fragments with trypsin and alpha-chymotrypsin differed from that of secretin. The fragments, S7-12 of P-1 and S13-21 of P-2 were determined to be different from the corresponding fragments obtained from secretin by HPLC analysis of their digestive fragments. The amino acid composition of each acid hydrolysate, following digestion with D-amino acid oxidase, was found to have less leucine or alanine content than secretin. The HPLC analysis of the fragments from P-1 and P 2 by tryptic and alpha-chymotryptic digestion showed that they are the same as those from synthetic D-Leu10 secretin or D-Ala17 secretin, respectively. Consequently, P-1 and P-2 are concluded to be the secretin diastereoisomers, D Leu10 and D-Ala17 secretin, respectively. PMID- 2565978 TI - AS and a 9.2 kb Pvu II class I HLA DNA restriction fragment. PMID- 2565979 TI - Genetic services in the context of DNA probes: what do they cost? AB - We describe results from the first year of a three year economic evaluation of genetic services in the context of DNA probes provided at three genetic centres in Great Britain. The analysis so far has concentrated on the costs of providing DNA diagnostic services. Estimates are given of the total costs of providing DNA probe services at each of the three centres, together with information for assessing the cost implications of future developments in these services. DNA probe services are likely to be funded as a Regional specialty. This paper concludes that relative to other developing medical technologies and as an investment by Regions, DNA probe services are inexpensive. An appraisal of the benefits to be derived from DNA probe services is still to be undertaken. PMID- 2565980 TI - Haplotypes identified by 10 DNA restriction fragment length polymorphisms at the human low density lipoprotein receptor gene locus. AB - Ten useful two allele restriction fragment length polymorphisms of the low density lipoprotein receptor gene were used for haplotype analysis in 45 unrelated familial hypercholesterolaemic (FH) patients, 60 normal controls, and 32 FH homozygotes, all of whom were white Afrikaners. Pedigree analysis in 27 informative heterozygous FH and 23 normal families has shown the segregation of at least 17 haplotypes in the normal population (111 chromosomes) compared to a predominant association of two of these haplotypes with the disease in the FH subjects. This association was further confirmed in 32 FH homozygotes, indicating at least two 'founder' members for the disease in the Afrikaner population. Recombination events were not detected in any of the families studied and we thus conclude that the haplotypes associated with FH function as specific markers for the disease and will allow presymptomatic diagnosis in affected families. PMID- 2565981 TI - Tolerance to nitroglycerin is caused by reduced guanylate cyclase activation. AB - Since the diminished vasodilatation characterizing tolerance to organic nitrates is associated with lower rises in 3', 5'-cyclic guanosine monophosphate (cGMP) levels, the possibility that nitrovasodilators desensitized guanylate cyclase (GC) when pre-incubated with coronary supernatants was studied. In the absence of cysteine, pre-incubation with nitroglycerin (NG) decreased GC-activity during subsequent incubation to 24 +/- 7% of control values, whereas six other nitrovasodilators had much smaller effects. When cysteine was present during pre incubation, NG-stimulation of GC remained significantly higher (59 +/- 3%; P less than 0.05), whereas the effects of other nitrovasodilators were not significantly changed. We also found that GC-activity, when reduced by pre-incubation with NG could only be restored by readdition of native coronary supernatant, suggesting that the enzyme became inactivated. NG pre-incubation of GC (in contrast to coronary strips) almost completely abolished the direct and thiol-independent stimulatory effect of 3-morpholinosydnonimine (SIN-1) down to 4.5 +/- 0.2%, whereas pre-incubation with other nitrovasodilators reduced the stimulatory response to SIN-1 to only 59 to 98%. Increasing concentrations of NG during pre incubation dose-dependently (IC50 = 0.13 mM) reduced the activating effect of SIN 1 during incubation. There was also a time dependence in NG-induced inactivation of GC which followed first order kinetics with a calculated half life of 2.5 min in the absence of a thiol. The latter was increased to 4.0 or 19.2 min, respectively, when glutathione or cysteine-methylester were present during pre incubation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2565982 TI - Glutamate degradation in the ischemic dog heart: contribution to anaerobic energy production. AB - The present study investigated the conversion of amino acids to succinate and the contribution of this pathway to anaerobic energy production during regional ischemia in the dog heart in situ. The relation between regional myocardial blood flow, estimated by the tracer microsphere technique, and myocardial contents of metabolites (glutamate, alanine, succinate, lactate) as well as their local arterio-venous differences (A-V) were determined. During 30 min of coronary artery occlusion, myocardial glutamate decreased from 2.3 mumol/g wet wt in control tissue to 1.2 mumol/g wet wt in severely ischemic areas, while aspartate was unaffected. Myocardial alanine increased in a 1: 1 stoichiometry compared to glutamate, and succinate accumulated. During control perfusion (118 mmHg), A-V of lactate, succinate and glutamate were +470, -0.7 and -3.9 nmol/ml, respectively. Stepwise reduction of perfusion pressure led to the release of lactate and succinate from the underperfused area; extraction of glutamate occurred at the lowest perfusion pressure investigated (34 mmHg; A-V: -500, -10.4 and +4.2 nmol/ml, respectively). The data indicate that during regional ischemia in vivo, succinate is synthetized exclusively from glutamate via 2-oxo-glutarate, following transamination with glycolytic pyruvate yielding alanine, while the contribution of aspartate is negligible. Using tissue levels of glutamate and lactate together with the local arterio-venous concentration differences of these compounds, it can be estimated that degradation of glutamate delivers 20% of the ATP generated by substrate level phosphorylation reactions. Thus energy production by the glutamate degradation pathway is significant in vivo under conditions of flow deprivation. PMID- 2565983 TI - Abuse of benzodiazepines. PMID- 2565984 TI - Drug and neurotransmitter receptors. New perspectives with clinical relevance. AB - Our understanding of drug and neurotransmitter receptors has been greatly advanced by receptor binding techniques. The theme of multiple-receptor subtypes with clinical relevance is highlighted by calcium antagonist and benzodiazepine receptors. Numerous drugs may exert therapeutic and untoward effects via calcium channels. Examples include effects of loperamide hydrochloride on diarrhea, the phenothiazine thioridazine hydrochloride on cardiac and ejaculatory function, and the neuromuscular toxicity of aminoglycosides. Elevated numbers of calcium antagonist receptors in hypertrophic cardiomyopathy may be involved in the pathophysiology of this disease. Benzodiazepines act via discrete central and peripheral receptors. At central receptors, benzodiazepines facilitate the inhibitory effects of the neurotransmitter gamma-aminobutyric acid. Peripheral benzodiazepine receptors are localized to mitochondria and enriched in several endocrine glands where they are influenced by therapeutic blood levels of benzodiazepines such as diazepam. PMID- 2565985 TI - Current concepts in brain resuscitation. AB - The area of cerebral resuscitation has become an exciting area of research in critical care medicine. It is a complicated field, however, which has seen attempts to protect the brain using a single therapy such as barbiturates ultimately disappoint investigators in the field. It is likely that much more work needs to be done in understanding the intracellular metabolic and biochemical effects of ischemia before therapies can be designed that are likely to be effective. This work might ultimately require knowledge of how ischemia or hypoxia interrupt cellular RNA and DNA machinery before these effective therapies can be developed. Before we are discouraged by the difficulty of the task, however, it is useful to review how much progress has been made in understanding the pathophysiology of ischemic brain injury in the past decade, so that we may be challenged to continue our efforts in this exciting area of critical care medicine. PMID- 2565986 TI - Intracellular distribution of cardiac beta-adrenoceptors in SHR and WKY. AB - To compare the intracellular distribution of beta-adrenoceptors in isolated myocytes of SHR with that of age-matched WKY and to examine changes in the distribution with aging, we measured the number of cell surface and total beta adrenoceptors in 6-(young groups) and 37-week old (old groups) SHR and WKY. The number of surface beta-adrenoceptors was significantly lower in 6-week old SHR compared with that in age-matched WKY. But there was no difference in the number of total beta-adrenoceptors between the two groups. The number of surface beta adrenoceptors in old groups was significantly reduced compared with that in the young groups, in both SHR and WKY. However, the number of total beta adrenoceptors in the old groups did not show any difference from that in the young groups. Isoproterenol-stimulated c-AMP formation in 6-week old SHR was significantly lower than that of WKY of the same age. Isoproterenol-stimulated c AMP formation in the old groups was significantly reduced than that of the young groups in both SHR and WKY. These results suggested that (1) there may be a difference in the intracellular distribution of cardiac beta-adrenoceptors between 6-week old SHR and WKY, (2) the intracellular distribution of cardiac beta-adrenoceptors may be changed with aging, and (3) intracellular distribution might be related to the difference in c-AMP formation to beta-agonist. PMID- 2565987 TI - [Evaluation of sialylated LewisX as a tumor-associated carbohydrate antigen in the sera of patients with gastric cancer]. AB - The clinical usefulness as a tumor-associated antigen of the new monoclonal antibody CSLEX1, which reacts with the sialylated LewisX antigen, has been evaluated serologically in 141 patients with a gastric cancer. The serum sialylated LewisX was measured by fluorescent EIA. The percent positive figure of these gastric cancer patients was 17.0% (24/141), which indicates a significant increase when compared with that of the controls. The percent-positive values, according to the clinical stage, were 9.2%, 9.1%, 15.6%, and 36.4% for stages I, II, III, and IV, respectively. The degree of liver metastasis and peritoneal dissemination of the cancer correlated with the percent-positive values of the antigen. This suggests that sialylated LewisX is useful as a tumor-associated antigen. PMID- 2565989 TI - [Prevention of ultraviolet insufficiency after a long stay by humans in an isolation chamber]. AB - The purpose of this study was to investigate biological effects of various regimens of ultraviolet irradiation in order to prevent UV insufficiency in men long kept in an enclosure. The subjects were kept for 60 days under artificial illumination in a Moscow laboratory in winter time. Two different regimens of UV irradiation were used and compared: 20 exposures at a dose of 0.75 MED and 20 exposures at incremental doses from 0.5 to 2 or 3 MED. Before and after 10 and 20 exposures (with medium-range radiation being predominant) the skin state, vitamin and mineral metabolism, and cytotoxicity of natural killers were investigated. The experimental observations allowed the conclusion that prophylaxis of UV insufficiency in men long kept in an enclosure can best be provided by UV irradiation at an incremental dose up to 2 MED. During this exposure skin sensitivity to UV radiation decreased, the content of vitamins D, A, E and that of Ca, P, Cu grew, and cytotoxicity of natural killers lightly increased. UV irradiation at a dose of 0.75 MED proved inadequate and that at a dose of 3 MED induced signs of excessive irradiation. PMID- 2565988 TI - [Rapid and complete hematopoietic reconstitution following peripheral blood stem cell autotransplantation in a child with T cell acute lymphoblastic leukemia]. AB - Rapid and complete hematopoietic reconstitution was achieved in a child with T cell acute lymphoblastic leukemia who was autografted with peripheral blood stem cells (PBSC). A large number of PBSC was collected by two courses of 3-4 hour lasting lymphopheresis during early remission induced by the second-line chemotherapy and then cryopreserved in liquid nitrogen. A myeloid progenitor cell dose of 203 X 10(4) CFU-GM/kg body weight was reinfused to the patient following marrow-ablative chemotherapy (MCNU 600 mg/m2, cytosine arabinoside 6 g/m2, etoposide 300 mg/m2, cyclophosphamide 160 mg/kg). Neutrophil count reached 0.5 X 10(9)/l by day + 7 and platelet count reached 20 X 10(9)/l by day + 9. Thereafter, white blood cell count continued to increase and reached a maximum of 38 X 10(9)/l on day + 14. Thus, the rapid recovery of hematopoiesis minimised marrow aplasia-related risks. This approach of stem cell rescue operation can be applied to the treatment of children with cancer, who otherwise have no hope to be cured, as an alternative to bone marrow transplantation. PMID- 2565990 TI - Serum interleukin-2 receptor: clinical and biological implications. PMID- 2565991 TI - [Topical beta adrenergic stimulants and respiratory arrest]. PMID- 2565992 TI - [Progress in the analysis of cystic fibrosis: prenatal diagnosis and carrier detection]. PMID- 2565993 TI - [Physiopathology of the transplanted heart. Review and clinical implications]. PMID- 2565994 TI - [Usefulness of a somatostatin analog in the treatment of chronic severe diarrhea caused by Cryptosporidium]. PMID- 2565995 TI - Non-invasive evaluation of hemodynamic, tremorogenic and biochemical effects in response to beta-adrenoceptor stimulation. AB - 1, 4, 12 and 24 micrograms SOM 1397 CL, a new beta 2-adrenergic bronchodilator, were administered by inhalation to 10 healthy volunteers in a double-blind, placebo-controlled, within-subject crossover study in order to assess circulatory, tremorogenic and biochemical effects. 1 microgram SOM 1397 CL did not cause any relevant changes in the measured parameters. After administration of 4 micrograms a slight but continuous increase in tremor amplitude and c-AMP was observed. The effects on hemodynamics and other laboratory values could be considered negligible. Overdoses of 12 and 24 micrograms resulted in a dose dependent increase in systolic blood pressure, pulse rate, tremor amplitude, alpha-AMP and lactic acid, as well as in a decrease of diastolic blood pressure and potassium. These effects may be partly attributable to beta 2-receptor stimulation. The method described in this paper can be applied as a useful tool for determination of beta 2-adrenergic activity in healthy volunteers, independent of the conventional methods which are used to evaluate efficacy of bronchodilators by the degree of bronchial muscular relaxation. PMID- 2565996 TI - Stereotactic fine-needle biopsy in 2594 mammographically detected non-palpable lesions. AB - To assess the accuracy of detection of breast cancers by mammography and stereotactic fine-needle biopsy (SFNB) 2594 mammographically detected non palpable lesions were sampled. On the basis of combined evaluation by mammography and cytology of these samples, 2005 (77.3%) of the cases were judged as benign lesions without need of surgery and only 1 of these turned out to be a cancer 14 months later. In 567 (21.9%) patients diagnostic and/or therapeutic surgery was done. Breast cancer was confirmed by histopathology in 429 (75.7%) of the patients operated on and a further 60(10.6%) had non-malignant pathological changes (eg, sclerosing adenosis, epitheliosis, fibroadenoma, or papilloma). Thus, surgery was justified in 86.3% (489) of the patients. In addition to the histopathologically verified cancers, another 22 (0.8%) breast cancers were diagnosed by mammography and cytology but these patients were not subjected to surgery for various reasons. A combination of mammography and SFNB offers a procedure of high sensitivity for early diagnosis of breast cancer. PMID- 2565997 TI - Effects of hydroxychloroquine and sulphasalazine on progression of joint damage in rheumatoid arthritis. AB - The effects of hydroxychloroquine and sulphasalazine on progression of joint damage shown by X-rays were compared in a double-blind, randomised trial in 60 patients with rheumatoid arthritis not previously treated with slow-acting antirheumatic drugs. X-rays of the hands and feet at the start and after 24 and 48 weeks of treatment were available for 28 patients treated with hydroxychloroquine and 22 treated with sulphasalazine. Erosions and joint space narrowing were scored by a single observer unaware of treatment. At baseline there were no significant differences in demographic, clinical, or radiographic characteristics between the treatment groups. Patients withdrawn because of lack of effect were included in the analysis. The median number of erosions was lower in the sulphasalazine than the hydroxychloroquine group at 24 weeks of treatment (2.5 vs 10) and the difference was significant at 48 weeks (5 vs 16; p less than 0.02). The difference in median total score of joint damage was significant at 24 weeks (6.5 vs 17; p less than 0.02) and at 48 weeks (8 vs 33; p less than 0.02). The increase in number of erosions and total score was significantly greater in the hydroxychloroquine than the sulphasalazine group, both after 24 weeks and after 48 weeks of treatment. PMID- 2565998 TI - Immunogenicity of a killed hepatitis A vaccine in seronegative volunteers. AB - 32 volunteers who lacked antibody to hepatitis A virus (HAV) received, at intervals of one month, three injections of a killed vaccine made from HAV propagated in diploid human fibroblast cell cultures. In 16 the vaccine was coupled with aluminium hydroxide as adjuvant. The serconversion rates measured by radioimmunoassay, with and without adjuvant respectively, were 13% and 25% at four weeks, 81% and 89% at eight weeks, and 94% and 100% at twelve weeks. Four weeks after the third injection all volunteers had high neutralising antibody titres. Twelve weeks later the titres of anti-HAV and anti-HAV neutralising antibodies had not declined. PMID- 2565999 TI - Warning symptoms of hypoglycaemia during treatment with human and porcine insulin in diabetes mellitus. AB - 32 subjects with long-term insulin-dependent diabetes mellitus (IDDM) were entered into a double-blind, randomised crossover trial with human and porcine insulin. They were treated during both periods with regular insulin and with protamine (NPH) insulin. 18 subjects started with human and 14 with porcine insulin; the two insulin periods each lasted twelve weeks; the insulin doses were much the same in the two periods (mean 23 [SD 9] U daily NPH; 14 [7] U daily regular insulin), as were blood glucose profiles and HbA1c values. There were 171 episodes of hypoglycaemia during human and 150 episodes during porcine insulin. Patients completed questionnaires after each hypoglycaemic episodes and at the end of the trial. Hunger and sweating without concomitant neuroglycopenic symptoms were significantly more frequent as initial warning symptoms during porcine than during human insulin (41% vs 20%), whereas neuroglycopenic symptoms were more frequent during human insulin. At the end of the trial 18 of 32 subjects reported diminished awareness of hypoglycaemia during human insulin compared with 6 of 32 during porcine insulin. Hypoglycaemia developed faster during human than during porcine insulin administration. The transfer of IDDM subjects from porcine to human insulin seems to alter warning symptoms of low blood glucose concentration, with consequent impairment of its early recognition. PMID- 2566000 TI - Relation between immunocytochemical estimation of oestrogen receptor in elderly patients with primary breast cancer and response to tamoxifen. AB - The relation between oestrogen receptor status of primary breast cancer as determined immuncytochemically on fine-needle aspirates and the response to tamoxifen therapy has been assessed in 52 patients. The proportion of cells staining correlated well with the likelihood of response, which emphasises the importance of tumour heterogeneity. PMID- 2566002 TI - A test object for assessing pulse oximeters. PMID- 2566001 TI - Clindamycin with primaquine for Pneumocystis carinii pneumonia. AB - Combined therapy with clindamycin and primaquine was used in twenty-eight episodes of Pneumocystis carinii pneumonia in 25 patients, of whom 17 had been unresponsive or intolerant to conventional treatment and 8 were being treated for the first time. The treatment was effective in all but two episodes, the main adverse reaction being a generalised maculopapular rash. PMID- 2566003 TI - Human papillomaviruses and the polymerase chain reaction. PMID- 2566004 TI - Metastatic fundamentals. PMID- 2566005 TI - Respiratory symptoms in farmers. PMID- 2566006 TI - Alpha-blockade for hypertension: indifferent past, uncertain future. PMID- 2566007 TI - Residual disability after ankle-joint injury. PMID- 2566008 TI - Hepatitis B vaccine in the expanded programme of immunisation: The Gambian experience. The Gambia Hepatitis Study Group. AB - As part of the Gambia Hepatitis Intervention Study, hepatitis B vaccine has been integrated into the national Expanded Programme of Immunisation (EPI) without major changes to the schedule of immunisation or to the mechanism of delivery. Serological results on a sample of vaccinated children at one year of age show that the strategy has been effective in reducing the prevalence of persistently infected children. The number of non-responders to the vaccine is low (less than 2%) and the antibody concentrations attained in responding children should give adequate protection when they are at high risk of persistent hepatitis B infection. Integration of hepatitis B vaccine into the EPI is feasible and effective in Africa. PMID- 2566009 TI - Cyclosporin in juvenile dermatomyositis. AB - Juvenile dermatomyositis in fourteen children who had not responded fully to steroids and other immunosuppressants and who had had chronic active disease for an average of 3 years was successfully treated with cyclosporin. Twelve patients had serious complications of the disease or of previous treatment. The response to cyclosporin included recovery of muscle strength and function and resolution of complications. It was possible to stop steroids or to reduce the steroid dose, which had previously been difficult, in all fourteen patients. In general, a low dose of cyclosporin (2.5-7.5 mg/kg daily) was sufficient and no serious side effects were seen. PMID- 2566010 TI - Obstacles to the treatment of detainees in South Africa. PMID- 2566011 TI - Fenoterol and fatal asthma. PMID- 2566012 TI - PAF antagonists in asthma. PMID- 2566013 TI - Human insulin hypoglycaemia unawareness. PMID- 2566014 TI - Latitude and heart disease. PMID- 2566015 TI - Liver transplantation in cystic fibrosis. PMID- 2566016 TI - Chest drain thrombectomy. PMID- 2566017 TI - Successful treatment of meningioma with octreotide. PMID- 2566018 TI - Vigabatrin in GABA metabolism disorders. PMID- 2566020 TI - Epstein-Barr virus associated B-cell lymphomas in AIDS and after organ transplantation. PMID- 2566019 TI - Detecting maternal cell contamination in prenatal diagnosis. PMID- 2566021 TI - Mitochondrial DNA mutation in family with Leber's hereditary optic neuropathy. PMID- 2566022 TI - Study of bivalent dengue vaccine in volunteers. PMID- 2566024 TI - Erythropoietin concentrations during treatment. PMID- 2566023 TI - Nasopharyngeal oxygen in children. PMID- 2566025 TI - Genetic differences between cystic fibrosis with and without meconium ileus. PMID- 2566026 TI - HTLV-I and neurological disease in South India. PMID- 2566027 TI - Health hazards on tap. PMID- 2566028 TI - International response to disasters. PMID- 2566029 TI - Self-prescribing and promotion of antidiarrhoeal drugs. PMID- 2566030 TI - British diplomas and limited registration. PMID- 2566031 TI - Origins of intravenous fluid therapy. PMID- 2566032 TI - Privacy for a handicapped child. PMID- 2566033 TI - When laser vaporisation for CIN fails, what next? PMID- 2566034 TI - AIDS and intranasal heroin. PMID- 2566036 TI - Reactivation of hepatitis B. PMID- 2566035 TI - Glove powder and HIV ELISA tests. PMID- 2566037 TI - Side-effects of subcutaneous apomorphine in Parkinson's disease. PMID- 2566038 TI - Symptomatic parvovirus B19 infection and heat-treated factor IX concentrate. PMID- 2566039 TI - Parenteral peptide supplements after major surgery. PMID- 2566040 TI - Diagnosis of superior sagittal sinus thrombosis by three-dimensional magnetic resonance flow imaging. PMID- 2566041 TI - Effect of a sunscreen in photosensitive patients. PMID- 2566042 TI - Conservative management of enterovirus infections in neonatal intensive care units. PMID- 2566043 TI - Senile dementia of Lewy body type and spectrum of Lewy body disease. PMID- 2566044 TI - Aspiration of boiling tea leading to respiratory failure. PMID- 2566046 TI - Medical quackbusters. PMID- 2566045 TI - Sterilisation of mentally handicapped woman approved by House of Lords. PMID- 2566047 TI - The death penalty and human rights. PMID- 2566048 TI - Double-masked trial of cyclosporin versus colchicine and long-term open study of cyclosporin in Behcet's disease. AB - The efficacy and safety of oral cyclosporin 10 mg/kg per day in Behcet's disease were compared in a randomised double-masked study with those of colchicine, 1 mg orally per day, and were also investigated in a long-term open study. The double masked study showed that cyclosporin was effective in treating not only the ocular manifestations of Behcet's disease but also oral aphthous ulcer, dermal lesions, and genital ulceration. Efficacy did not weaken during long-term treatment. PMID- 2566049 TI - Retrovirus-like sequences in Graves' disease: implications for human autoimmunity. AB - On Southern blotting of DNA extracted from thyroid glands of five patients with Graves' disease, two probes (720 bp and 942 bp) for gag human immunodeficiency virus type 1 (HIV-1) gave a positive hybridisation signal in all samples tested. DNAs from peripheral blood mononuclear cells hybridised with the 720 bp gag HIV-1 probe in three of the five patients, none of whom had antibodies to HIV-1. Negative results were obtained with DNA from normal thyroid glands, thyroid neoplasms, various unrelated normal tissues, and virus-infected human cell lines. The intensity of the signal and the pattern of bands observed with the DNA of Graves' patients were heterogeneous and, in general, were not the same in the thyroid glands and peripheral blood mononuclear cells of individual patients. Similarly, no correlation was found between the positive hybridisation signals and other genetic and immunological indices or the duration of anti-thyroid drug treatment at the time the patients were investigated. The findings suggest the presence of a novel retrovirus, and the retrovirus-like sequences seem to be closely associated with thyroid autoimmunity. PMID- 2566050 TI - Ten-year follow-up study of islet-cell antibodies and childhood diabetes mellitus. AB - To find out whether subclinical autoimmunity precedes onset of nonfamilial insulin-dependent diabetes mellitus (IDDM), 4806 schoolchildren aged 5-19 years from a township in Holland were followed-up for at least ten years after blood was sampled for measurement of islet-cell antibodies (ICA). ICA positivity conferred a relative risk of IDDM of 533 (95% CI 145-1955). In the 10 years of follow-up 4 of the 8 ICA-positive subjects became insulin dependent, whereas the probability of being free of IDDM was 99.9% for those who were ICA-negative at the start of the study. The findings suggest that, although chronic autoimmunity involving the pancreatic beta-cells may precede non-familial IDDM by many years, a positive ICA test on a single occasion predicts the development of IDDM in only 4 out of 8 subjects over a period of 10 years. PMID- 2566051 TI - Usefulness of ophthalmoscopy in mild to moderate hypertension. AB - A panel of two physicians and two ophthalmologists examined 25 patients with untreated essential hypertension by direct ophthalmoscopy and assessment of fundal photographs; daytime ambulatory sphygmomanometric blood pressure monitoring, estimation of left ventricular mass by electrocardiography and two dimensional echocardiography, and measurement of urinary microalbumin excretion were also carried out. No relation was found between blood pressure determined by clinic or ambulatory sphygmomanometry and retinopathy. The retinal features sought on fundal photographs were the percentages of arteriovenous crossings with venule nipping, venule deviation, or attenuation of venular light reflex. The ratio of arteriolar to venular diameter was measured. Only focal narrowing of arterioles was associated with higher blood pressure. There was no independent relation between retinal features and age, measures of left ventricular mass, or urinary microalbumin excretion. Assessment of arteriovenous crossing abnormalities by direct ophthalmoscopy was subject to wide variability among the panel members. Direct ophthalmoscopy was not clinically useful in the assessment of mild to moderate hypertension, whereas urinary microalbumin excretion correlated strongly with clinic blood pressure. PMID- 2566053 TI - AIDS: prevention, policies, and prostitutes. PMID- 2566052 TI - Femoral artery recanalisation with percutaneous angioplasty and segmentally enclosed plasminogen activator. AB - To establish whether re-occlusion of the femoral artery could be prevented, in 6 consecutive patients undergoing percutaneous transluminal angioplasty (PTA) for superficial femoral artery occlusion the recanalised segment was isolated, with a 7-French double-balloon catheter. 5 mg recombined human tissue-type plasminogen activator (rt-PA) and 1000 IU heparin were then infused into the enclosed space for 30 minutes, followed by intravenous heparin for 24 hours. At 10 and 30 days all 6 patients had evidence of recanalisation and remission of symptoms. Mean ankle-arm pressure index improved by 72% at 24 hours, by 118% at 10 days, and by 103% at 30 days after the procedure. No patients had major complications. Treatment of superficial femoral artery occlusions by PTA with rt-PA and intravenous heparin seems to prevent rethrombosis. PMID- 2566054 TI - Diabetic neuropathy. PMID- 2566055 TI - Reperfusion or persisting occlusion? PMID- 2566056 TI - Peers reviewed. PMID- 2566057 TI - New development in phototherapy. PMID- 2566058 TI - Evaluation of four screening tests for bacteriuria in elderly people. AB - Four screening tests for bacteriuria were assessed at ward level in 418 elderly subjects and were compared with standard methods of bacterial culture. The tests were visual appearance; microscopy; dipstick for nitrite, leucocyte esterase, protein, and blood; and dipstick for nitrite and organisms. The sensitivity of the tests varied from 85.6% to 98.3%, and the specificity from 18.4% to 82.9%. A combination of visual appearance and dipstick testing for nitrite and leucocyte esterase gave a sensitivity of 96.1% with a specificity of 50.6%, and could have reduced by almost one-third the number of urine samples submitted to the laboratory for processing. PMID- 2566059 TI - Transplant or chemotherapy in acute myelogenous leukaemia. International Bone Marrow Transplant Registry. AB - It is not known whether chemotherapy or bone-marrow transplantation is the more effective treatment for acute myelogenous leukaemia (AML) in first remission. Consequently, some centres recommend transplants only for patients for whom a poor response to chemotherapy is predicted. To assess how reasonable this strategy is, data from 704 recipients of HLA-identical sibling transplants for AML in first remission were analysed. 5-year leukaemia-free survival (LFS) was 48% (95% confidence interval 43-53%); leukaemia relapse rate was 20% (16-24%). Improved LFS was associated with younger age and lower white blood cell count at diagnosis. These prognostic variables are similar to those reported to affect outcome after chemotherapy. These findings suggest that a strategy of treatment assignment based on risk factor analysis is unlikely to resolve the controversy of transplant versus chemotherapy for AML in first remission. PMID- 2566060 TI - Cachectin/tumour necrosis factor. PMID- 2566061 TI - Who needs steroid receptor assays? PMID- 2566062 TI - Medical education: seat of pants versus structured flying. PMID- 2566063 TI - Fluconazole versus ketoconazole in oropharyngeal candidiasis in AIDS. PMID- 2566064 TI - Detecting t(14;18) in paraffin-embedded lymphoma tissue. PMID- 2566065 TI - Polymerase chain reaction to confirm extranodal progression of follicular lymphoma. PMID- 2566067 TI - Brush cytology for colorectal cancer. PMID- 2566066 TI - Papillomavirus infection and skin cancer in renal allograft recipients. PMID- 2566068 TI - Glaucoma and race. PMID- 2566069 TI - Dimethylsulphoxide-induced encephalopathy. PMID- 2566070 TI - Cardiac hypersensitivity to 5-aminosalicylic acid. PMID- 2566071 TI - Nifedipine and impaired oxygenation in patients with chronic bronchitis and cor pulmonale. PMID- 2566072 TI - Nifedipine in the treatment of Raynaud's disease in childhood. PMID- 2566073 TI - Low-dose zidovudine for AIDS. PMID- 2566074 TI - HIV-2 in West Africa in 1966. PMID- 2566075 TI - HTLV-I antibodies in Papua New Guinea. PMID- 2566076 TI - Drug advertising. PMID- 2566077 TI - Coma scale to predict survival. PMID- 2566078 TI - Mental tests on elderly patients. PMID- 2566079 TI - Cyclophosphamide in severe polymyositis. PMID- 2566080 TI - EDRF and microvascular constriction after angioplasty. PMID- 2566081 TI - Ivermectin and prothrombin time. PMID- 2566082 TI - 99mTc-HMPAO uptake and washout in stroke. PMID- 2566083 TI - Fail-safe mechanism against autoimmunity. PMID- 2566084 TI - Cerebral consumption of glucose. PMID- 2566085 TI - Polyarticular gout. PMID- 2566086 TI - Liver transplantation in primary hyperoxaluria type 1. PMID- 2566087 TI - Hepatotoxicity of paracetamol in combination with interferon and vinblastine. PMID- 2566088 TI - Fatal adult respiratory distress syndrome after quinine overdose. PMID- 2566089 TI - Missed diagnosis of mycobacterial infection. PMID- 2566090 TI - Low frequency of erythromycin-resistant Streptococcus pneumoniae in The Netherlands. PMID- 2566091 TI - Salmonella infection and ulcerative colitis. PMID- 2566092 TI - Anaerobiospirillum: cause of a "new" zoonosis? PMID- 2566094 TI - Latitude and ischaemic heart disease. PMID- 2566093 TI - Diet, pre-eclampsia, and intrauterine growth retardation. PMID- 2566095 TI - Transdermal testosterone. PMID- 2566096 TI - Cardiac hypertrophy in old age. PMID- 2566097 TI - Exercise training, fitness, and asthma. PMID- 2566098 TI - Monoclonal technique to aid decision on endocrine therapy in breast cancer. PMID- 2566099 TI - Total serum IgE levels in children with pertussis. PMID- 2566101 TI - AIDS in the UK and world wide. PMID- 2566100 TI - The Baltimore dispute. PMID- 2566102 TI - Hemodynamic effects of hypertonic saline in the conscious rat. AB - The present study examines the role of vasopressin and the sympathetic nervous system on the hemodynamic effects of an infusion of hypertonic saline (NaCl 1.5 M) in conscious rats. The cardiovascular response to hypertonic saline was similar in both untreated and hexamethonium-pretreated rats. Mean arterial pressure increased by 15 mmHg as a consequence of the elevation of total peripheral resistance, while cardiac index was decreased. The administration of an antagonist to the pressor activity of vasopressin in rats with intact reflexes, partially decreased mean arterial pressure and total peripheral resistance and increased cardiac index toward basal values. In contrast, the hemodynamic response to hypertonic saline was totally reverted when the vasopressin antagonist was injected in the hexamethonium-pretreated rats. The results of the present study indicate that the hypertensive response induced by hypertonic saline in conscious rats is due to the vasoconstrictor effects of both vasopressin and the sympathetic nervous system. PMID- 2566103 TI - Inhibition of rat colonic motility and cardiovascular effects of new gut-specific beta-adrenergic phenylethanolaminotetralines. AB - We have investigated the ability of the new putative beta-adrenergic agonists phenylethanolaminotetralines (PEAT) to inhibit intestinal motility in relation to their cardiovascular effects, in ethylurethane-anesthetized rats. The representative PEAT SR 58375A, SR 58572A and SR 58539B and the reference beta adrenergic agonists isoproterenol, salbutamol and ritodrine caused dose-related inhibition of proximal colon spontaneous motility: ED50 (microgram/kg, i.v.) 210, 92 and 19; 5.6, 176 and 201, as listed. This inhibition was prevented by the beta adrenergic antagonist alprenolol, but not by desipramine (which prevented the inhibition of colonic motility by tyramine and enhanced that by norepinephrine). The minimal effective doses (MED) of isoproterenol, salbutamol and ritodrine raising heart rate and/or lowering blood pressure (by 10 to 20%), was substantially lower (about 1/10 to 1/150) than their ED50 for inhibition of colonic motility. The MED raising heart rate of the three PEAT, on the other hand, was about twice (SR 58375A and SR 58572A) to five (SR 58539B) times their ED50 for inhibition of colonic motility. None of the PEAT lowered blood pressure up to the top tested dose. Therefore the PEAT may prove preferable to the currently best tolerated beta-adrenoceptor agonists, because they appear less liable to induce cardiovascular side effects. This supports the prospective therapeutic interest of PEAT for intestinal hypermotility disorders. PMID- 2566104 TI - Regional distribution and regulation of preprosomatostatin messenger RNA in the striatum, as revealed by in situ hybridization histochemistry. AB - Quantitative in situ hybridization was used to examine the regional distribution of preprosomatostatin messenger RNA (PPSOM mRNA) in the dorsal striatum (caudo putamen) of the mouse. In addition, because mesencephalic dopaminergic neurons project to the striatum where they play a role in the regulation of peptide containing neurons, the effect of dopamine receptor blockade on the levels of striatal PPSOM mRNA was determined. Sagittal brain sections from male Swiss Webster mice were processed for in situ hybridization histochemistry using an 35S radiolabelled RNA probe in order to quantify levels of PPSOM mRNA in individual neurons of the dorsal caudo-putamen using light microscopy and computer-assisted grain analysis. In control animals, individual neurons of the dorsolateral caudo putamen had higher levels of PPSOM mRNA than did those of the medial part of the structure. Treated mice were injected with fluphenazine-N-mustard (FNM), an antagonist which, at the dose used (4 mumol/kg), irreversibly blocks dopamine D2 but not D1 receptors in the mouse striatum. FNM treatment (for 2 days, twice a day) produced an increase in striatal dopamine turnover and a decrease in PPSOM mRNA levels in the lateral, but not the medial striatum. The results indicate that there is a lateral to medial gradient in the levels of PPSOM mRNA per individual neuron in the dorsal caudo-putamen of control animals, which is abolished by FNM treatment. This suggests that intrinsic striatal somatostatinergic neurons are differentially regulated by dopamine, depending on their lateromedial location within the striatum. PMID- 2566106 TI - Critical evaluation of bioluminescence methods for quantifying bacterial adhesion to polystyrene. AB - A total of 134 strains of Escherichia coli which included 60 fecal and 74 urinary isolates, cultured in liquid and on solid media, were examined for adhesive properties using bioluminescence and haemagglutination methods. The study aimed to verify whether irrespective of the absence or presence of flagella, there is any relation between haemagglutination and bioluminescence test. Examining the results we failed to note any correlation between the two methods: strains bearing MS, MR or MS-MR adhesins adhered to polystyrene at random. Even though it is fast and easy to perform, bioluminescence is not an alternative to traditional methods to reveal MS adhesins. PMID- 2566105 TI - Restriction-map variation at the zeste-tko region in natural populations of Drosophila melanogaster. AB - Restriction-map variation in 64 X chromosome lines extracted from three different natural populations of Drosophila melanogaster was investigated with seven six nucleotide-recognizing enzymes for a 20-kb region including the zeste and tko genes. Ten restriction-site and four length polymorphisms (two insertions and two deletions) were detected. Contrary to the predicted lower level of variation for genes on the X chromosome, the level of variation attributable to nucleotide substitution (estimated heterozygosity/nucleotide = 0.004) was similar to that previously reported for autosomal loci. The amount of insertion/deletion variation in the studied region was within the range observed in autosomal regions and thus not explainable by a simple selection model against the effects of insertional mutations. A general lack of linkage disequilibrium between polymorphic sites was observed. PMID- 2566108 TI - Japanese encephalitis after a two-week holiday in Bali. AB - Japanese encephalitis is described in a 10-year-old girl after a short holiday in Bali. Four days after returning to Australia the patient presented with a high fever, stupor and rapidly-developing focal neurological signs. Recovery occurred gradually over a period of three months and she has returned to school. Japanese encephalitis viral infection was confirmed by a marked rise in specific haemagglutination-inhibition antibodies and the presence of immunoglobulin M antibodies to the flavivirus group. It is important to be aware of the possibility of arboviral infection in patients with encephalitis. In view of the recent outbreaks of Japanese encephalitis in Asia, travellers to the region should be warned to protect themselves from mosquito-bites. PMID- 2566107 TI - Lipids and lipoproteins in patients undergoing coronary-artery surgery. AB - Fasting blood samples were obtained from 290 patients who were undergoing elective coronary-artery graft procedures, and cholesterol, triglyceride and high density lipoprotein cholesterol levels were measured. The 1983 National Heart Foundation of Australia's Risk Factor Prevalence Study was used as a source of age- and sex-matched "control" data. Of these patients, 80% had cholesterol levels of greater than 5.5 mmol/L; in 55% of patients, the level exceeded 6.5 mmol/L. Only 4% of patients who received a graft showed hypertriglyceridaemia alone (triglyceride level, greater than 2 mmol/L). Combined hyperlipidaemia (cholesterol level, greater than 5.5 mmol/L and triglyceride level, greater than 2.0 mmol/L) was present in 52% of subjects. Low-density lipoprotein cholesterol levels exceeded 3.5 mmol/L in 69% of men and in 71% of women. In terms of five 10 year age intervals, mean plasma triglyceride and cholesterol levels were elevated significantly in patients who had undergone a coronary-artery grafting procedure compared with those of subjects in the National Heart Foundation study. The mean high-density lipoprotein cholesterol levels were markedly-lower compared with those of the subjects in the National Heart Foundation study. Of those patients whose plasma cholesterol levels were less than 5.5 mmol/L, 97% of patients had high-density lipoprotein cholesterol levels that were less than the mean level for subjects in the National Heart Foundation study. Thus, a very-high proportion of patients who underwent coronary-artery bypass surgery had lipid abnormalities which required intervention postoperatively. PMID- 2566109 TI - Postoperative analgesia. PMID- 2566111 TI - [A 35-year-old patient with necrotizing exanthema, restrictive cardiomyopathy, liver cirrhosis and acute psychosis]. PMID- 2566110 TI - Insect repellents. PMID- 2566112 TI - [Central effects of five beta-adrenergic receptor blockers in healthy volunteers: a quantitative EEG study]. AB - The effects of five beta blockers on the central nervous system of healthy subjects was studied by computerized EEG analysis. All subjects underwent continuous recording with a Holter magnetic type recorder during the experimental period. For 10 consecutive days, five groups of subjects received alternately placebo and the beta blockers acebutolol 600 mg, carteolol 20 mg, metoprolol 200 mg, pindolol 30 mg and sotalol 320 mg. EEG recordings (C4/P4, P4/02 and C3/P3, P3/01) lasting 5 min were made between 8.30 and 9.30 a.m. Subjects were at rest with eyes closed and there was no vigilance control. The signal was recorded on a magnetic tape recorder and then processed by Nicolet MED 80 system. Comparisons of absolute and relative powers and of average frequencies were then made between the different sequences and groups. The possible correlations between the changes observed in the power spectrum and the clinical, pharmacological and pharmacokinetic specific properties of each beta blocker are discussed. PMID- 2566114 TI - An Eco RI polymorphic site in the human complement C4 gene distinguishes juvenile rheumatoid arthritis (JRA) susceptibility-bearing haplotypes. AB - Susceptibility to acquire Juvenile Rheumatoid Arthritis (JRA) is linked to HLA DR5 and DRw8 antigens in Caucasoid populations. However, the frequency of HLA-DR5 is too high in the normal Spanish population and JRA cannot thus be found to be associated with this antigen. It has been found a 14.3 kb-C4-Eco RI restriction fragment length polymorphism which correlates significantly with JRA and may be used as a marker for this disorder in Spaniards. PMID- 2566113 TI - A Thy-1 negative lymphoma cell variant defective in the formation of glycosyl phosphatidylinositol membrane protein anchors. AB - Thy-1 is a glycoprotein present on the membrane of murine cells of the T-lineage. The mature Thy-1 is anchored to the membrane via a glycolipid, phosphatidylinositide. In order to study the regulation of the synthesis and membrane insertion of this protein, the biochemical properties of a Thy-1.2 negative variant T-lymphoma cell (RL male 1.4) were studied. It contains intracellular Thy-1 protein but fails to express it on the cell surface. While the wild type and the mutant show similar labelling of the intracellular Thy-1 glycoprotein with amino acids, no ethanolamine is incorporated into the Thy-1 molecule of RL male 1.4. A plasmid, pT1, containing the normal Thy-1.2 gene and bacterial gpt gene was transfected into RL male 1.4 and into the murine plasmacytoma cell, J558L. A transfected plasmacytoma, T1J2, synthesized a normal sized Thy-1 protein and displayed the antigen on the membrane. In contrast, the mycophenolic acid resistant RL male 1.4 transfectants did not display Thy-1.2 on the cell surface, despite the presence of substantial amounts of Thy-1 intracellularly. Two other antigens known to be anchored in the membrane by phospholipid, Ly-6e and Qa-2, were also examined in RL male 1.4. RL male 1.4 did not express Ly-6e after alpha interferon induction. In addition, the expression of Qa-2 antigen was greatly diminished in RL male 1.4 in comparison to RL male 1.3. Thus, the defect in RL male 1.4 is not restricted to Thy-1.2, but includes other similarly anchored glycoproteins as well. This implies that the addition of phospholipid to core proteins is similar, if not identical, for all these molecules and that the RL male 1.4 cell lacks the capacity to from the lipid glycoprotein linkage required for the expression of these proteins on the cell surface. PMID- 2566115 TI - Neurotransmitter systems and receptor plasticity in brain aging. PMID- 2566116 TI - A mitochondrial DNA mutation as a cause of Leber's hereditary optic neuropathy. AB - Leber's hereditary optic neuropathy is a maternally inherited disease associated with the late onset of bilateral loss of central vision and cardiac dysrhythmias. The maternal inheritance is explained by the mitochondrial origin of the disease. Analysis of the sequence of a mitochondrial DNA has indicated that a single nucleotide change at position 11778 is associated with this disease. This mutation converts the 340th amino acid of NADH dehydrogenase subunit 4 from an arginine to a histidine and eliminates an SfaNI endonuclease restriction site. A survey of restriction-fragment-length polymorphisms in the mitochondrial DNA of three independent families with this disease (an American black and two white European families) and 10 controls confirmed that this SfaNI site is associated with the disease. A phylogenetic tree for mitochondrial DNA polymorphism and sequence variants from three probands with Leber's disease and four controls was constructed, and the mutation at position 11778 was found to be associated with two mitochondrial DNA backgrounds--an American black mitochondrial DNA and a European mitochondrial DNA. Thus, this mutation must have arisen twice independently. Since the mutation correlated with symptoms of Leber's disease in both cases, these findings indicate that the mutation is a cause of the disease. This genetic analysis has identified the specific point mutation in the mitochondrial DNA that results in Leber's hereditary optic neuropathy. PMID- 2566117 TI - Amyloid A4 protein and its precursor in Down's syndrome and Alzheimer's disease. AB - In patients with Alzheimer's disease, amyloid fibrils that are aggregates of A4 protein subunits are deposited in the brain. A similar process occurs at an earlier age in persons with Down's syndrome. To investigate the deposition of amyloid in these diseases, we used a radioimmunoassay to measure levels of the amyloid precursor (PreA4) in the serum of 17 patients with Down's syndrome, 15 patients with Alzheimer's disease, and 33 normal elderly controls. The mean (+/- SD) concentration of serum PreA4 was increased 1.5-fold in patients with Down's syndrome (2.49 +/- 1.13 nmol per liter) as compared with that in controls (1.68 +/- 0.49 nmol per liter; P less than 0.007); the levels in patients with Alzheimer's disease were similar to those in controls (1.83 +/- 0.78; P less than 0.98). We also found that the concentration of PreA4 in the brain tissue of two adults with Down's syndrome (100 and 190 pmol per gram) was higher than that in the brain tissue of either 26 patients with Alzheimer's disease (64.4 +/- 17.3 pmol per gram) or 17 elderly controls with neurologic disease (68.5 +/- 26.3 pmol per gram). Immunocytochemical studies of brain tissue from 26 patients with Down's syndrome showed that the deposition of A4 protein amyloid began in these patients approximately 50 years earlier than it began in 127 normal aging subjects studied previously, although the rate of deposition was the same. We conclude that, since the gene for PreA4 is on the long arm of chromosome 21, which is present in triplicate in Down's syndrome, overexpression of this gene may lead to increased levels of PreA4 and amyloid deposition in Down's syndrome. However, since increased levels of PreA4 are not present in Alzheimer's disease, additional factors must account for the amyloid deposition in that disorder. PMID- 2566118 TI - Cytoadherence of knobless Plasmodium falciparum-infected erythrocytes and its inhibition by a human monoclonal antibody. AB - Red blood cells infected with mature stages of the malaria parasite Plasmodium falciparum bind to the endothelial lining of capillaries and venules. This sequestration is important for the survival of the parasite but may have severe consequences for the host. For example, it is involved in the causation of cerebral malaria which carries 25% mortality. Knob-like protrusions present on the surface of infected erythrocytes have been considered necessary but not sufficient for this cytoadherence. Here we describe the adhesion to endothelial cells of infected erythrocytes which do not have knobs. A human monoclonal antibody (33G2) which was specific for an epitope containing regularly spaced dimers of glutamic acid present in the repeated amino-acid sequences of some defined P. falciparum antigens was found to inhibit cyto-adherence and may therefore be an important reagent for elucidating the molecular basis of parasite sequestration. PMID- 2566119 TI - Where next with peer-review? PMID- 2566121 TI - Regulating the cell cycle. PMID- 2566120 TI - Suppressor effects and cyclic AMP accumulation by the CD29 molecule of CD4+ lymphocytes. AB - Integrins are a superfamily of related molecules whose function, where known, is to mediate adhesion. The so-called very-late-activation antigen (VLA) family includes at least five distinct heterodimers, each composed of a unique alpha subunit non-covalently associated with a common beta-subunit. Several members of the family have been shown to bind extracellular matrix proteins, but the function of VLA-4 is so far unknown. VLA-4 is the only member of the family detected on thymocytes and resting T cells. We show here that an antibody which recognizes the beta-subunit of VLA-4 (CD29) on T cells can inhibit CD4+ cell proliferation triggered by CD2 or CD3, and that binding of this antibody to activated T cells leads to an increase in cyclic AMP levels which is comparable to that elicited by forskolin. These negative signalling effects are unique to this antibody: other CD29 antibodies do not affect the growth of activated CD4 cells but enhance the proliferation of whole T cell populations and abrogate the suppressive effects of mitomycin-treated CD8 cells on CD4-cell growth. Taken together, our results indicate that VLA-4 functions in cell-cell interactions and that it is the target for the suppressive effects of CD8 cells on CD4 cells. PMID- 2566122 TI - Evidence from mitochondrial DNA that African honey bees spread as continuous maternal lineages. AB - African honey bees have populated much of South and Central America and will soon enter the United States. The mechanism by which they have spread is controversial. Africanization may be largely the result of paternal gene flow into extant European populations or, alternatively, of maternal migration of feral swarms that have maintained an African genetic integrity. We have been using both mitochondrial and nuclear DNA restriction fragment length polymorphisms to follow the population dynamics between European and African bees. In earlier reports, we suggested that if African honey bees had distinctive mitochondrial (mt) DNA, then it could potentially distinguish the relative contributions of swarming and mating to the Africanization process. Because mtDNA is maternally inherited, it would not be transmitted by mating drones and only transported by queens accompanying swarms. Furthermore, the presence of African mtDNA would reflect unbroken maternal lineages from the original bees introduced from Africa. The value of mtDNA for population studies in general has been reviewed recently. Here we report that 19 feral swarms, randomly caught in Mexico, all carried African mtDNA. Thus, the migrating force of the African honey bee in the American tropics consists of continuous African maternal lineages spreading as swarms. The mating of African drones to European queens seems to contribute little to African bee migration. PMID- 2566123 TI - Neotropical Africanized honey bees have African mitochondrial DNA. AB - Non-indigenous African honey bees have invaded most of South and Central America in just over 30 years. The genetic composition of this population and the means by which it rapidly colonizes new territory remain controversial. In particular, it has been unclear whether this 'Africanized' population has resulted from interbreeding between African and domestic European bees, or is an essentially pure African population. Also, it has not been known whether this population expanded primarily by female or by male migration. Restriction site mapping of 62 mitochondrial DNAs of African bees from Brazil, Venezuela and Mexico reveals that 97% were of African (Apis mellifera scutellata) type. Although neotropical European apiary populations are rapidly Africanized by mating with neotropical African males, there is little reciprocal gene flow to the neotropical African population through European females. These are the first genetic data to indicate that the neotropical African population could be expanding its range by female migration. PMID- 2566124 TI - [Strong inhibition of gastric acid secretion: advantages and possible hazards]. PMID- 2566125 TI - [Is strong inhibition of gastric acid in the treatment of peptic ulcer sensible?]. PMID- 2566126 TI - [Drug treatment of reflux esophagitis: effects of strong gastric acid inhibition]. PMID- 2566127 TI - [Slow-acting antirheumatic agents]. PMID- 2566128 TI - [A testis tumor in the left adnexi]. AB - Cryptorchidism increases the incidence of malignant testicular tumours. The case is described of a normally functioning male patient with a tumour in an undescended testis, in whom a special type of intersexuality was found: the persistent mullerian duct syndrome, one of the types of male pseudohermaphroditism, in which no disturbances of virilisation occur and which is often associated with cryptorchidism and other developmental anomalies. PMID- 2566130 TI - Regulatory activity and topological distribution of folliculo-stellate cells in rat anterior pituitary cell aggregates. AB - An enriched population of cells immunoreactive to antiserum against S-100 protein, a marker of folliculo-stellate (FS) cells in the rat pituitary, was obtained by separation of dispersed pituitary cells from adult female rats by gradient sedimentation at unit gravity. The effect of FS cells on the stimulation and inhibition of prolactin (PRL) and growth hormone (GH) release was studied by coaggregation experiments of the FS cell-enriched population with respectively a lactotroph-enriched and a somatotroph-enriched population from adult female rats. The FS cell population not only attenuated the stimulation of PRL and GH release, but also significantly attenuated the inhibition of PRL release by 10, 30 or 300 nM dopamine (DA), and the inhibition of GH release by 0.1 nM somatostatin (SRIF). The stimulatory action of angiotensin II (AII) on PRL secretion in the presence of DA was also attenuated by the FS cells. Light microscopic evaluation of immunostained semithin sections showed a meshwork of cytoplasmic extensions of FS cells as well as follicular structures in the aggregates. There was no preferential association of FS cells with certain cell types. The permeability of the aggregates to diffusing molecules was tested at the ultrastructural level by the lanthanum hydroxide tracing technique. Lanthanum traced the intercellular gaps over the entire aggregate irrespective of whether the proportional number of FS cells was high or low, indicating that FS cells do not seal off certain areas in the aggregate by the formation of tight junctions. It is suggested that FS cells attenuate the action not only of stimulatory but also inhibitory secretagogues on hormone-secreting pituitary cells. The possible physiological relevance of the present findings is supported by the topological distribution of the FS cells in the aggregates, which closely resembles that of the intact pituitary. PMID- 2566129 TI - Possible involvement of beta endorphin(1-31) and dynorphin(1-13) in the central hypotensive mechanism of action of alpha methyldopa. AB - The present study was performed to gain more information on the nature of the opioid peptide(s) involved in the mechanism of action of alpha-methyldopa. Conscious, normotensive Wistar rats were used and all treatments were given intracisternally. For blood pressure and heart rate, pretreatment with a midportion beta-endorphin antiserum resulted in a parallel shift to the right of the dose-response curve for alpha-methyldopa. In addition, when rats were pretreated with various dilutions of this antiserum and treated with a constant dose of alpha-methyldopa, the antiserum dose-dependently inhibited alpha methyldopa-induced hypotension and bradycardia. Using antisera specifically recognizing the C-terminus of beta-, gamma- and alpha-endorphin, respectively, revealed that only the beta-endorphin antiserum inhibited the decrease in blood pressure seen after administration of alpha-methyldopa. An antiserum against [Met5]enkephalin did not influence the cardiovascular responses following alpha methyldopa. On the other hand, a dynorphin(1-13) antiserum also inhibited in a dose-dependent manner the hypotension induced by alpha-methyldopa. When administered 3 h after the injection of alpha-methyldopa, the beta-endorphin and dynorphin(1-13) antisera failed to reverse the hypotension induced by alpha methyldopa. The results favor a role for beta-endorphin(1-31) and dynorphin(1-13) in the hypotension centrally mediated by alpha-methyldopa. PMID- 2566131 TI - Presence of a particulate thyrotropin-releasing hormone-degrading pyroglutamate aminopeptidase activity in rat liver. AB - The data of this work describe the presence in the rat liver of a thyrotropin releasing hormone (TRH)-degrading particulate metal-containing pyroglutamate aminopeptidase of high molecular weight. Following the fractionation of liver homogenate in 0.25 M sucrose, solubilization of the particulate fraction with papain and gel filtration on ACA34, an enzyme activity was detected which converts TRH into pyroglutamic acid and histidyl-proline diketopiperazine (cyclo[His-Pro]; cHP). [L-Histidine-2,5-3H]TRH and [L-proline-2,3-3H]TRH were used as a tracer. Products formed were separated by thin-layer chromatography, localized and quantified by scanning for radioactivity. Enzyme activities were tested using preferential site-directed inhibitors. Particulate pyroglutamate aminopeptidase activity was found to be sensitive to chelating agents. The physicochemical properties of this particulate aminopeptidase were distinct from the soluble pyroglutamate aminopeptidase from the same source. The particulate enzyme shared several similarities with particulate pyroglutamate aminopeptidase from adenohypophysis, brain and with serum enzyme, reported to have narrower specificity for TRH compared to the soluble pyroglutamate aminopeptidase from several tissues. The Km of the gel-filtrated enzyme is 27 microM and the specific activity 306 pmol.min-1.mg protein-1. Although no definite role has been established for this enzyme, it might be a potential determinant of cHP concentrations in liver. Furthermore, cHP is known to possess biological activities and specific binding sites in liver membranes. One of the major sites for TRH breakdown in vivo, the liver, probably represents a target tissue for TRH, especially for pancreatic TRH, and the particulate enzyme involved in the conversion of TRH into cHP may assume a specific function. PMID- 2566132 TI - Effect of CH-38083, a selective antagonist of alpha 2-adrenoceptors, on renal sympathetic function. AB - The effects of 7,8-(methylenedioxi)-14-alpha-hydroxyalloberbane HCl (CH-38083), a structurally new, selective and potent alpha 2-adrenoceptor antagonist, and of idazoxan, were studied on both the spontaneous activity of the postganglionic sympathetic renal nerve and on the clonidine- or xylazine-induced inhibitory action in anaesthetized cats. The drug CH-38083 (30-200 micrograms/kg, i.v.) caused a sustained increase of the sympathetic activity and blood pressure. Larger doses of idazoxan (200-500 micrograms/kg, i.v.) were needed to induce similar effects. Both antagonists were effective in antagonizing the sympatho inhibitory effect of clonidine or xylazine. The excitatory response to selective alpha 2-adrenoceptor antagonists suggests that the sympathetic output undergoes tonic inhibition due to a permanent alpha 2-adrenoceptor stimulation. PMID- 2566133 TI - [Surgical therapy of duodenal ulcer today]. PMID- 2566134 TI - [Cryptorchism. Clinical considerations and therapy]. PMID- 2566135 TI - [Recent progress in dopaminergic control of aldosterone secretion]. AB - The role of dopamine on the aldosterone secretion-regulation mechanism has been confirmed by the identification of dopaminergic receptors on adrenal glomerulosa cells. Though the renin-angiotensin system plays an important primary role in the regulation of aldosterone secretion it has recently been suggested that dopamine has direct inhibitory influence on aldosterone release. Dopamine has no effect on basal plasma aldosterone levels, while it inhibits the aldosterone response in a sodium-depleted state or upright posture. Furthermore, in vitro studies have shown that dopamine modulates the activity of zona glomerulosa enzymes regulating the biosynthesis of aldosterone. The use of dopamine agonist and antagonist substances has shown that dopamine inhibits aldosterone biosynthesis by activating DA2 dopaminergic receptors identified in adrenal glomerulosa cells. PMID- 2566136 TI - Peripheral inflammation is associated with increased dynorphin immunoreactivity in both projection and local circuit neurons in the superficial dorsal horn of the rat lumbar spinal cord. AB - The present study combined the retrograde transport of fluorescent tracers with the immunocytochemical identification of dynorphin A(1-8) in superficial dorsal horn neurons to examine whether peripheral inflammation-induced dynorphin increases are found in local circuit neurons only or also in neurons projecting at least to the caudal mesencephalon. Evidence is presented that complete Freund's adjuvant-induced inflammation produces a large increase in the number of lamina I dynorphin-containing projection and non-projection neurons, and in the number of lamina II dynorphin local circuit neurons. PMID- 2566137 TI - Abnormal expression of tyrosine hydroxylase-like immunoreactivity in intraocular transplants of rat caudate nucleus. AB - The purpose of the present study was to examine tyrosine hydroxylase (TH)-like immunoreactivity in single intraocular grafts of caudate nucleus, and in caudate grafts which were co-grafted with substantia nigra. Grafts of caudate obtained from fetal rats (crown-rump length: 15-17 mm) were placed into the anterior chamber of eye of adult female Sprague-Dawley rats, and were allowed to survive from 6 to 20 months. The host rats were then perfused with fixative and the tissue was prepared for immunocytochemistry for TH. In 7 out of 8 surviving caudate grafts, including those containing no substantia nigra, tyrosine TH-like immunoreactive (THLI) cell bodies were present. The results of our study suggest that TH is inappropriately expressed in caudate grafts placed in the anterior chamber of the eye. PMID- 2566138 TI - Tyrosine hydroxylase-immunoreactive intrinsic neurons in the Auerbach's and Meissner's plexuses of humans. AB - We carried out an immunohistochemical study of the Auerbach's and Meissner's plexuses of the human alimentary tract, using antiserum against tyrosine hydroxylase (TH), a rate-limiting enzyme of the catecholamine-synthesizing pathway. TH-immunoreactive intrinsic neuronal cell bodies were observed in both plexuses in almost all parts of the alimentary tract, being most frequent in the Auerbach's plexus of the lower esophagus. PMID- 2566139 TI - Somatostatin cyclic octapeptide analogs which preferentially bind to SOMa receptors block a calcium current in rat superior cervical ganglion neurons. AB - To characterize further the somastatin (SOM) receptor mediating Ca2+ current reduction in rat superior cervical ganglion (SCG) neurons, the effects of three synthetic SOM octapeptide analogs, D-Phe-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH2 (IM-4 82), D-Nal-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH2 (DC 13-116), and D-Phe-Cys-Phe-D-Trp Lys-Thr-Cys-Thr-OL (SMS 201-995), which bind preferentially to pituitary SOM receptors (SOMa) were investigated. Ca2+ currents were recorded using the whole cell variant of the patch-clamp technique from neurons isolated enzymatically from adult rat SCG. Application of the SOM analogs (0.003-3 microM) produced a rapid, reversible, and concentration-dependent decrease in Ca2+ current amplitude in addition to slowing the rising phase of the Ca2+ current. Estimates of the concentration producing half-maximal block (EC50) and maximum attainable block (Bmax) for DC 13-116, IM 4-28, and SMS 201-995 were 196, 67, and 9.5 nM, respectively, and 52, 57, and 48%, respectively. The results suggest that the SOM receptor on SCG neurons more closely resembles the SOMa receptor of the anterior pituitary than the SOMb receptor of cerebral cortical membranes. PMID- 2566140 TI - Kynurenate and 2-amino-5-phosphonovalerate interact with multiple binding sites of the N-methyl-D-aspartate-sensitive glutamate receptor domain. AB - By studying the binding of [3H]glycine and [3H]glutamate to rat synaptic membranes in the presence of 2-amino-5-phosphonovalerate (APV) and kynurenate (KYN) we have demonstrated that KYN is more potent than APV in displacing [3H]glycine, while an opposite order of potency was seen in displacing [3H]glutamate. Moreover, 2-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP) inhibited only [3H]glutamate binding. The [3H]MK-801 specific binding was inhibited by all of the above antagonists; this action was abolished by glutamate, while glycine partially reversed only the action of KYN. Hence, KYN inhibits glutamate receptors by preferentially interfering with glycine recognition sites, while APV preferentially interacts with N-methyl-D-aspartate (NMDA) recognition sites. PMID- 2566141 TI - Distribution of dynorphin B and methionine-enkephalin in the mouse hippocampus: influence of genotype. AB - Immunohistochemical techniques were used to localize dynorphin B and methionine enkephalin in the mouse hippocampus. Methionine-enkephalin-like immunoreactivity was found within the somata of interneurons distributed mainly in and around the CA1 stratum pyramidale and stratum granulosum as well as in the mossy fibers. Dynorphin B appeared to be confined to the mossy fiber pathway. In addition, we observed a difference between the inbred mouse strains DBA/2 and C57BL/6 with regard to the areas of the dynorphinergic mossy fiber projections: the intra- and infrapyramidal terminal fields were larger in the latter group. PMID- 2566143 TI - Open Forum for Health Information of New Zealand Association Inc. PMID- 2566142 TI - Activation of N-methyl-D-aspartate receptors contributes to the EPSP at perforant path synapses in the rat dentate gyrus in vitro. AB - The excitatory postsynaptic potential (EPSP) evoked in the granule cells of the rat dentate gyrus following low frequency stimulation of the perforant path has been investigated using intracellular recording. The EPSP was reduced by low microM concentrations of the non-N-methyl-D-aspartate (NMDA) receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). A small CNQX-resistant component of the EPSP remained. This could be blocked by the NMDA receptor antagonist (+/-)-2 amino-5-phosphonovalerate, was enhanced in Mg2+-free medium and showed a potential-dependency characteristic of the activation of NMDA ionophores. These results demonstrate that NMDA receptors contribute to the EPSP in the granule cell. PMID- 2566144 TI - Loss of heterozygosity at the c-raf locus in small cell lung carcinoma. AB - The c-raf-1 oncogene is located at chromosome 3p25, near a region known to be specifically deleted in patients with renal cell carcinoma and small cell lung carcinoma (SCLC). From cytogenetic analyses of SCLC cell lines, we have estimated that one c-raf-1 allele was deleted in approximately 80% of the cases. However, c raf-1 was generally thought to be distal to the most common deletion in SCLC, 3p14-23. Using restriction site polymorphisms (RFLPs) located within the c-raf-1 locus, we have examined DNA from 84 human lung carcinomas. In an analysis of 11 paired (normal versus tumor) SCLC DNA samples, all five informative cases showed loss of heterozygosity at this locus in the corresponding tumor sample. Analysis of 73 unpaired lung carcinoma DNAs showed that out of 31 non-SCLC samples, 19% were heterozygous for the BglI polymorphism and 25% showed heterozygosity with TaqI. However, all of the 42 SCLC samples were homozygous for both of these RFLPs. This striking loss of heterozygosity at the c-raf-1 locus in SCLC indicates that one allele of c-raf-1 is deleted in SCLC. The kinase activity of the c-raf protein appears to be constitutively activated in these cells. Whether this apparent activation results from genetic or epigenetic events is under investigation. PMID- 2566145 TI - Allelic variation of the c-raf-1 proto-oncogene in human lymphoma and leukemia. AB - Several lines of evidence point to the involvement of the proto-oncogene c-raf-1 in the development of lymphoma and leukemia and in a previous study we found variants of this gene in 3 lymphoma patients. To see whether variation at this locus plays a role in predisposition to these cancers, we have examined the frequency of an unusual EcoRI allele of c-raf-1 in 99 non-Hodgkin's lymphoma and 28 acute lymphoblastic leukemia patients, and in 182 controls. To optimize the chance of detecting a difference, we selected 113 of the controls from geriatric patients with no family history of cancer. No difference in the frequency of the allele was found between patients and controls, thereby refuting our hypothesis that this polymorphism of the c-raf-1 locus contributes to genetic susceptibility to these two related cancers. We estimate that the frequency of the rarer b EcoRI allele of this locus is 0.091 +/- 0.012 in the Australian Caucasian population. PMID- 2566146 TI - The oncogene associated with human papillary thyroid carcinoma (PTC) is assigned to chromosome 10 q11-q12 in the same region as multiple endocrine neoplasia type 2A (MEN2A). AB - In this report we assigned to chromosome 10q the human oncogene PTC frequently associated with the papillary type of thyroid carcinoma. Using an informative panel of human-mouse somatic cell hybrids and 'in situ' hybridization to human metaphase chromosomes, we localized the PTC gene at bands q11-q12 of chromosome 10. These bands belong to one of the two regions on chromosome 10 linked to the cancer syndrome multiple endocrine neoplasia type 2A (MEN2A). Therefore, it is suggested that genes clustered in certain regions of chromosome 10 could be involved in the developmental regulation of the thyroid gland. PMID- 2566148 TI - [Neurohumoral mechanisms of regulating the motor-evacuatory function of the stomach]. PMID- 2566147 TI - Bladder-neck opening test in spinal cord injury patients using a new i.v. alpha blocking agent, alfuzosin. AB - A bladder-neck opening test using alfuzosin, a new alpha-adrenoceptor blocking agent, was carried out in 21 patients with spinal cord injury. The efficacy of alfuzosin was assessed with 4 simple urodynamic parameters: micturition, residual urine, posterior urethral pressure and diameter. Both mean urethral pressure and diameter were significantly affected after the administration of 5 mg i.v. alfuzosin. The test was clinically positive in 18 patients: 11 out of 13 patients using intermittent catheterisation or continuous drainage urinated and 6 out of 8 patients already tapping had reduced residual urine volumes. A decrease in posterior urethral pressure was also observed in 2 out of 3 patients who did not respond clinically to alfuzosin. Alfuzosin was well tolerated during this test. Oral alfuzosin should therefore be investigated in patients who gave a satisfactory response to such a test. PMID- 2566149 TI - Prostaglandins and neurotransmission at the guinea pig and rabbit urinary bladder. AB - High concentrations of prostaglandins (PGE1, PGE2, or PGE2 alpha) (2 x 10(-6) M) produced a slow contraction of longitudinal strips of detrusor muscle taken from the bladders of guinea pigs and rabbits. At a lower concentration (10(-6) M) prostaglandins enhanced contractions produced by field stimulation of nerves in guinea pig but not rabbit strips. The contractions were not affected by indomethacin. Contractions of guinea pig strips in response to acetylcholine at 10(-4) M were enhanced by prostaglandins and unaffected by indomethacin. Membrane potentials of smooth muscle cells recorded with micro electrodes, were unchanged up to 10(-6) M PGE2. Above this the cells were depolarized with an increase in frequency of spontaneous action potentials. Synchronous recording of electrical and mechanical activity with the double sucrose gap indicated a decrease in amplitude of the evoked excitatory junction potential and action potential even when the contraction was enhanced in the presence of PGE2. Responses to repeated stimulation at 10 Hz for 1 min were progressively depressed. This trend was slightly reduced by PGE2 but unaffected by indomethacin. It is concluded that prostaglandins are not normally released by the nerves to the urinary bladder but are able to facilitate contraction in the guinea pig. This effect is probably on the excitatory-contraction coupling, possibly by mobilizing Ca2+. Some modification of transmitter release by the nerves may also occur. PMID- 2566150 TI - Nucleotide sequence of the F41 fimbriae subunit gene in Escherichia coli B41. PMID- 2566151 TI - Six RFLPs for human T cell receptor alpha (TCRA) on chromosome 14. PMID- 2566152 TI - An HaeIII RFLP for the human homeo box-containing gene EN1. PMID- 2566153 TI - An SstI RFLP for the human homeo box-containing gene EN2. PMID- 2566154 TI - PvuII and RsaI RFLPs for the human homeo box-containing gene EN2. PMID- 2566155 TI - An StuI RFLP at the human cholesteryl ester transfer protein (CETP) locus. PMID- 2566156 TI - A restriction fragment length polymorphism at murine Glud locus co-segregates with Rib-1, Es-10, and Tcra on chromosome 14. PMID- 2566157 TI - An EcoRI polymorphism of the human von Willebrand factor cDNA (VWF). PMID- 2566158 TI - An anonymous single-copy clone, p30-1-60, identifies a frequent RFLP on chromosome 3p [HGM9 no. D3S86]. PMID- 2566159 TI - A polymorphic locus at Xq27-28 detected by the probe U6.2 [DXS304]. PMID- 2566160 TI - [Effectiveness of a new beta 2-agonist (broxaterol) on bronchospasm induced by methacholine (blind study versus salbutamol)]. AB - A new beta 2-agonists, broxaterol, administered by oral spray has been tested to evaluate its action modifying on the bronchial hyperreactivity to metacholine. Such drug has been administered double blind Vs salbutamol individualizing for both, at 15' and 90' premedication the PD20 FEV1 (doses of metacholine necessary to reduce of 20% the FEV1). The results that we have obtained confirm the protective capacity for both drugs on the bronchoconstriction induced by aspecific stimulus. However, in confront with salbutamol, the broxaterol presents an action more lasting in the time. Such requisite and absence of collateral effects permit to affirm that this new beta 2-agonists is fit for its utilization in the infantile intrinsic asthma. PMID- 2566162 TI - [Systemic effects of beta-blockaders in eyedrops]. PMID- 2566161 TI - [Effects of xamoterol in moderate cardiac insufficiency]. AB - Xamoterol, a partial agonist of beta 1-adrenoceptors, was tested as a cardiotonic drug in 10 patients with moderate chronic heart failure. The trial was double blind drug versus placebo for 3 months and open for one year. At 3 months the patients were clinically improved with downgrading by one NYHA class, and there was a significant increase in response to a 36-second exercise, as compared with the placebo group. Haemodynamic index, a 14 per cent fall in pulmonary wedge pressure and a 7 per cent increase in left ventricular stroke work as compared with the placebo group. This clinical and haemodynamic improvement was paralleled by a reduction of the double product on exercise. In long term use, xamoterol did not seem to induce tachyphylaxis, and few side-effects were recorded during this trial. PMID- 2566163 TI - [Dermatitis and hepatitis in a child treated with salazosulfapyridine in hemorrhagic rectocolitis]. PMID- 2566164 TI - Regulation of the gastrin promoter by epidermal growth factor and neuropeptides. AB - The regulation of gastrin gene transcription was studied in GH4 pituitary cells transfected with constructs comprised of the first exon of the human gastrin gene and various lengths of 5' regulatory sequences ligated upstream of the reporter gene chloramphenicol acetyltransferase. Gastrin reporter gene activity in GH4 cells was equal to the activity of a reporter gene transcribed from the endogenously expressed growth hormone promoter. The effect of a variety of peptides on gastrin gene transcription including epidermal growth factor (normally present in the gastric lumen), gastrin-releasing peptide, vasoactive intestinal peptide, and somatostatin (present in gastric nerves) was assessed. Epidermal growth factor increased the rate of gastrin transcription almost 3 fold, whereas thyrotropin-releasing hormone and vasoactive intestinal peptide increased gastrin transcription 2- and 1.5-fold, respectively. Gastrin-releasing peptide, a peptide that strongly stimulates gastrin release, weakly increased gastrin transcription (1.3-fold). Somatostatin inhibited the increase in gastrin transcription induced by epidermal growth factor, thyrotropin-releasing hormone, and vasoactive intestinal peptide. Constructs containing various lengths of 5' regulatory sequences defined a response element -40 to -82 base pairs (bp) 5' to the transcription initiation site. This 40-bp sequence contains Sp1 and AP2 binding sites, which suggests that epidermal growth factor and thyrotropin releasing hormone stimulate gastrin gene transcription through transcription factors that bind to Sp1 and/or AP2 motifs. PMID- 2566166 TI - Inhibition of insulin receptor phosphorylation by peptides derived from major histocompatibility complex class I antigens. AB - Peptides from the alpha 1 region (residues 61-85) of the D and K molecules of the major histocompatibility complex class I antigens inhibit insulin-induced tyrosine kinase activity of the purified human insulin receptors (IRs) as measured both by autophosphorylation and IR-mediated substrate [poly(Glu,Tyr)] phosphorylation. Half-maximal effect of the Dk-(61-85) peptide on IR autophosphorylation is obtained at 1.2 microM, and almost complete inhibition of IR kinase activity is obtained at 10 microM peptide. The corresponding K kappa (61-85) peptide has a significantly weaker effect on autophosphorylation. No such effects are observed with nine peptides of similar length, but unrelated to major histocompatibility complex class I antigens. Neither of the major histocompatibility complex class I-derived peptides has any effect on the constitutively active kinase of a genetically engineered cytoplasmic IR domain. Further, insulin binding to IR is unaltered in the presence of the major histocompatibility complex class I-derived peptides. The inhibitory activity of the peptides on insulin-induced IR phosphorylation facilitated the observation that IRs require insulin to become substrate for an independent tyrosine kinase. In the presence of an inhibitory peptide, the constitutively active cytoplasmic IR kinase domain only phosphorylates the intact IR in the presence of insulin. We conclude that the tyrosine kinase activity of IRs may be altered by peptide interaction at an allosteric site and, moreover, IRs require insulin to assume a conformation permitting phosphorylation by an independent kinase. PMID- 2566165 TI - Changes in stability and allosteric properties of aspartate transcarbamoylase resulting from amino acid substitutions in the zinc-binding domain of the regulatory chains. AB - Changes in subunit interaction energies linked to the allosteric transition of the regulatory enzyme aspartate transcarbamoylase (ATCase; EC 2.1.3.2) from Escherichia coli are localized in part at interfaces between the six catalytic (c) and six regulatory (r) polypeptide chains. Site-directed mutagenesis has been used to construct enzymes with amino acid substitutions in a limited region of the zinc-binding domain of the r chains. Substitution of Ser or His for r114 Cys, one of four cysteines binding the structural zinc ion in the regulatory chain, leads to incorrectly folded chains as shown by the inability to detect stable assembled holoenzyme in cell extracts. Replacement of r111 Asn by Ala at the interface between an r chain and a c chain in the apposing catalytic trimer causes a complete loss of the homotropic and heterotropic effects characteristic of wild-type ATCase. Moreover, sedimentation velocity experiments demonstrated that this mutant enzyme exists in the R ("relaxed") conformation in the absence of active site ligands due to preferential destabilization of the T ("taut") conformation relative to the R state. In contrast, replacement of r113 Asn by Ala at the interface between adjacent r and c chains leads to an increase in the cooperativity of the enzyme. When r139 Lys is replaced by Met, Vmax is reduced by 50% compared to wild-type ATCase, whereas it is increased about 2-fold when r142 Glu is replaced by Asp. Amino acid substitutions in this domain significantly affect subunit interaction energy as measured by rate of subunit exchange when holoenzymes are incubated with isolated catalytic subunits, thus permitting measurements of the effect of the bisubstrate analog N-(phosphonacetyl)-L asparatate in weakening intersubunit interactions. Subunit exchange increased about 9-fold for the r142 Glu----Asp mutant and almost 20-fold for the r142 Glu-- -Ala mutant in the presence of the ligand. PMID- 2566167 TI - Gene for proliferating-cell nuclear antigen (DNA polymerase delta auxiliary protein) is present in both mammalian and higher plant genomes. AB - Proliferating-cell nuclear antigen (PCNA; also called cyclin) was originally described in proliferating mammalian cells as a nuclear protein with an apparent Mr of 33,000-36,000 and recently was found to be a DNA polymerase delta auxiliary protein. To elucidate whether PCNA/cyclin is a universal protein necessary for proliferation of eukaryotes, a search was conducted for PCNA/cyclin homologues in higher plants. In Southern blot-hybridization analysis, a rat PCNA/cyclin cDNA probe hybridized with homologous sequences in genomic DNAs from rice, soybean, and tobacco. A PCNA/cyclin-related molecular clone (pCJ-1) was isolated from rice DNA and was partially sequenced. The pCJ-1 probe hybridized with a 1.2-kilobase transcript in RNA from rice root tips and shoots. Immunoblot analysis of the soluble extract of soybean root tips with monospecific anti-PCNA/cyclin identified an immunoreactive protein with an apparent Mr of 34,000. Immunohistochemical analysis revealed the presence of an immunoreactive PCNA/cyclin protein in the nuclei of cells in the meristem of soybean root tips. The highly homologous nature of the gene for PCNA/cyclin throughout the animal and plant kingdoms suggests that the product of the gene plays an essential role in DNA replication in eukaryotes. PMID- 2566168 TI - Tumor-specific loss of 11p15.5 alleles in del11p13 Wilms tumor and in familial adrenocortical carcinoma. AB - We have compared constitutional and tumor genotypes in nine cases of hereditary Wilms tumor (WT) and in three unrelated cases of familial adrenocortical carcinoma (ADCC). Since susceptibility to these tumors can be observed in malformation syndromes associated with a constitutional deletion of band 11p13 (WT) and with a constitutional duplication of band 11p15.5 (WT, ADCC), we investigated these two candidate regions by using 11p polymorphic markers. As expected, somatic chromosomal events, resulting in a loss of heterozygosity limited to region 11p15.5, were observed in the tumor of two familial cases of adrenocortical carcinoma. Surprisingly, however, analysis of the WT of two patients with a constitutional deletion of band 11p13, associated with aniridia, genitourinary abnormalities, and mental retardation (WAGR syndrome), revealed a loss of heterozygosity limited to region 11p15.5. These data therefore suggest that observation of a specific loss of heterozygosity may not necessarily point to the site of the initial germinal mutation. Together with previous similar observations of a loss of heterozygosity limited to 11p15.5 in breast cancer and in rhabdomyosarcoma, our data suggest that region 11p15.5 may carry a non-tissue specific gene that could be involved in genetic predisposition, in tumor progression, or in both. PMID- 2566169 TI - Virus-induced delayed-type hypersensitivity reaction is sequentially mediated by CD8+ and CD4+ T lymphocytes. AB - After subcutaneous inoculation into the hind foot of a mouse, the lymphocytic choriomeningitis (LCM) virus multiplies locally, attaining 10(7)-10(8) mouse infectious units per g of tissue; elimination commences around day 7. About 1 day earlier, the foot begins to swell, which is regarded as a delayed-type hypersensitivity (DTH) reaction. To answer the question of whether the local inflammatory response is involved in virus clearance, we needed to known what cells mediate both these phenomena. With three different procedures--namely, depletion in vivo of defined cells by treatment of mice with monoclonal antibodies ("serologic surgery"), adoptive immunization with negatively selected cells, and adoptive immunization with cells from mice differing at the major histocompatibility gene complex--it is shown that the LCM virus-induced local DTH reaction consists of two phases that are sequentially mediated by (first) class I restricted cytotoxic/suppressive CD8+ and (second) class II-restricted helper/inducer CD4+ T lymphocytes. In contrast, for virus elimination only the former subset of T lymphocytes was found to be needed. Thus, an association may exist between the CD8+ cell-mediated component of the local DTH response and control of the infection, but the CD4+ cell-mediated part appears to be of doubtful antiviral relevance. PMID- 2566170 TI - Specific interaction of aurintricarboxylic acid with the human immunodeficiency virus/CD4 cell receptor. AB - The triphenylmethane derivative aurintricarboxylic acid (ATA), but not aurin, selectively prevented the binding of OKT4A/Leu-3a monoclonal antibody (mAb) and, to a lesser extent, OKT4 mAb to the CD4 cell receptor for human immunodeficiency virus type 1 (HIV-1). The effect was seen within 1 min at an ATA concentration of 10 microM in various T4+ cells (MT-4, U-937, peripheral blood lymphocytes, and monocytes). It was dose-dependent and reversible. ATA prevented the attachment of radiolabeled HIV-1 particles to MT-4 cells, which could be expected as the result of its specific binding to the HIV/CD4 receptor. Other HIV inhibitors such as suramin, fuchsin acid, azidothymidine, dextran sulfate, heparin, and pentosan polysulfate did not affect OKT4A/Leu-3a mAb binding to the CD4 receptor, although the sulfated polysaccharides suppressed HIV-1 adsorption to the cells at concentrations required for complete protection against HIV-1 cytopathogenicity. Thus, ATA is a selective marker molecule for the CD4 receptor. ATA also interfered with the staining of membrane-associated HIV-1 glycoprotein gp120 by a mAb against it. These unusual properties of a small molecule of nonimmunological origin may have important implications for the study of CD4/HIV/AIDS pathogenesis and possibly treatment. PMID- 2566171 TI - Co-release of acetylcholine and gamma-aminobutyric acid by a retinal neuron. AB - Rabbit retinas were vitally stained with 4',6-diamidino-2-phenylindole (DAPI), a fluorescent compound that selectively accumulates within the cholinergic amacrine cells. The retinas were then incubated in vitro in the presence of radioactive gamma-aminobutyric acid (GABA) and autoradiographed. The cells that accumulated DAPI were found to accumulate GABA, confirming immunohistochemical evidence that the cholinergic amacrine cells contain GABA. Incubation of retinas in the presence of elevated concentrations of K+ caused them to release acetylcholine and GABA, and autoradiography showed depletion of radioactive GABA from the cholinergic amacrine cells. This indicates that the cholinergic amacrine cells can secrete acetylcholine and GABA. Retinas were double-labeled with [14C]GABA and [3H]acetylcholine, allowing simultaneous measurement of their release. The release of [14C]GABA was found to be independent of extracellular Ca2+. Radioactive GABA synthesized endogenously from [14C]glutamate behaved the same way as radioactive GABA accumulated from the medium. In the same experiments the simultaneously measured release of [3H]acetylcholine was strongly Ca2+-dependent, indicating that the releases of acetylcholine and GABA are controlled by different mechanisms. Synaptic vesicles immunologically isolated from double labeled retinas contained much [3H]acetylcholine and little or no [14C]GABA. These results suggest that the cholinergic amacrine cells release acetylcholine primarily by vesicle exocytosis and release GABA primarily by means of a carrier. PMID- 2566173 TI - T cells as helpers and suppressor cells in immunity. PMID- 2566174 TI - Alternative pathways for T lymphocyte activation. PMID- 2566172 TI - Single potassium channels opened by opioids in rat locus ceruleus neurons. AB - Currents through single-ion channels were recorded in the cell-attached configuration from locus ceruleus neurons enzymatically dissociated from newborn rats. When the selective mu opioid receptor agonist Tyr-D-Ala-Gly-MePhe-Gly-ol was in the patch-clamp electrode, unitary inward currents were observed with conductance of approximately 45 pS (measured at zero pipette potential, with 150 mM potassium in the recording electrode). Long silences, lasting many seconds to minutes, separated periods of activity of similar durations. Within such activity periods the distribution of closed times of the channels was best fitted by the sum of two exponential functions (time constants approximately 1 and 30 ms), and the durations of channel openings were fit by a single exponential function; mean open time increased from 2 to 120 ms as agonist concentration increased. Channel activity was not seen when high concentrations of opioids were applied to the neuron outside the patch-clamp recording electrode, indicating intimate coupling between receptor and potassium channel. Unitary currents with similar properties were also seen when pipettes contained alpha 2 adrenoceptor agonists or somatostatin. Taken with previous findings, the results indicate that mu opioid receptors, alpha 2 adrenoceptors, and somatostatin receptors can couple directly to membrane potassium channels through the local intermediary action of a GTP binding protein. PMID- 2566175 TI - Contrasuppression, tolerance, and tumor immunity. PMID- 2566176 TI - Prevention of tumor growth and enhancement of cell-mediated immunity by an antigen-specific contrasuppressor factor from a T cell hybridoma. AB - We have produced a contrasuppressor T cell hybridoma which has positive effects on multiple forms of cell-mediated immunity. First of all, it protects the adoptive transfer of contact sensitivity from suppressor cells and factors. In addition, TcsF modifies the response to normally tolerogenic administrations of hapten, leading mice to develop contact sensitivity and CTL activity instead of tolerance. Most relevant to this conference, mice which have been both treated with AF5.C6 TcsF and painted with TNCB resist challenge with highly malignant TNP modified tumors. These experiments suggest a decisive role for contrasuppression in tumor rejection. PMID- 2566178 TI - Novel peripheral neurotransmitters in invertebrates. PMID- 2566177 TI - Low doses of chemotherapy to inhibit suppressor T cells. PMID- 2566179 TI - The polymorphic oxidation of beta-adrenoceptor antagonists. PMID- 2566180 TI - Benzodiazepine abuse and dependence in psychiatric inpatients. AB - Over a period of five and a half years, 792 inpatients with benzodiazepine (BZD) abuse and dependence in accordance with the WHO definition were registered at a university psychiatric hospital. One-quarter of them abused BZD exclusively, while three-quarters suffered from polytoxicomania or were alcohol dependent as well. It was possible to distinguish two groups of patients: one with primary, mainly low-dose, dependence preferring lorazepam and the other with secondary, often high-dose, dependence with diazepam as the main drug. In 108 patients with isolated BZD dependence, withdrawal symptoms of somatic, psychological, or perceptual quality were observed. The severity of the withdrawal syndrome seemed to depend on the time of consumption, dose, mode of withdrawal, and type of BZD compound. Following abrupt cessation, 11 patients with long-standing dependence on high BZD doses developed withdrawal psychoses, presenting a delirious, paranoid-hallucinatory, or depressive-anxious syndrome. PMID- 2566181 TI - Relations between psychotic symptoms and serum prolactin levels. AB - Constantly high prolactin values during treatment with neuroleptics at least contribute to the continuance of pathological psychic symptoms. Single-drug therapy should always be the aim in long-term treatment of chronic schizophrenic patients, gynaecological disorders should be recognised in time, and an adenoma of the pituitary should be considered in differential diagnosis if the prolactin levels are permanently increased. PMID- 2566182 TI - The influence of exaprolol upon the ischaemic rat heart and its interaction with sarcolemmal (Na+ + K+)-ATPase. AB - The capability of cyclohexylphenol exaprolol of protecting the ischaemic myocardium during ischaemic cardiac arrest was assessed in the isolated working rat heart. Exaprolol added to the perfusion medium in a dose of 10(-7) mol.l-1 only minimally influenced the left ventricular function (reduced the stroke volume by 18.84% and cardiac output by 14.63%). The hearts were subjected to global ischaemia for 75 min at 26 degrees C and subsequently reperfused for 60 min at 37 degrees C. The recovery of left ventricular function following reperfusion, expressed as a percentage of preischaemic functional performance was used as an indicator of the ischaemic tolerance of the heart. The effect of exaprolol on sarcolemmal (Na+ + K+)-, Mg2+- and Ca2+-ATPase activities was also examined. Exaprolol-pretreated hearts revealed better postischaemic recovery of the left ventricular dP/dt max and stroke volume as well as improved efficiency in the transformation of chemical energy to mechanical work. Exaprolol in 10(-4) mol.l-1 concentration significantly stimulated the specific activity of sarcolemmal (Na+ + K+)-ATPase. Possible mechanisms of the salutary effect of exaprolol on the ischaemic heart are discussed. PMID- 2566183 TI - From chlorpromazine to tardive dyskinesia (brief history of the neuroleptics). PMID- 2566184 TI - The psychodynamic action of psychopharmacologic drugs and the target symptom versus the anti-psychotic approach to psychopharmacologic therapy: thirty years later. PMID- 2566185 TI - Pharmacokinetic study of iminodibenzyl antipsychotic drugs, clocapramine and Y 516 in dog and man. AB - The pharmacokinetic properties of the iminodibenzyl antipsychotic drugs clocapramine (CCP, 3-chloro-5-[3-(4-carbamoyl-4-piperidino piperidino) propyl] 10, 11-dihydro-5H-dibenzo[b, f]azepine) and Y-516 (3-chloro-5-[3-(2-oxo-1, 2, 3, 5, 6, 7, 8, 8a-octahydroimidazo [1,2-a] pyridine-3-spiro-4'-piperidino) propyl] 10, 11-dihydro-5H-dibenzo[b, f]azepine) were investigated in dog and man. Dogs were administered CCP and Y-516 intravenously, intraperitoneally, and orally, and the concentrations of the parent drugs and their metabolites in the plasma and urine were determined. Half-life (t1/2) was approximately the same by all three administration routes, being approximately 5 h for CCP and 3 h for Y-516. Bioavailability following oral administration was 0.16 +/- 0.01 (mean +/- SD, n = 3) for CCP and 0.29 +/- 0.07 for Y-516. The fractions of dose absorbed following oral administration were 0.43 +/- 0.07 and 0.79 +/- 0.24, and the fractions of dose metabolized in the liver due to the first-pass effect were 0.63 +/- 0.05 and 0.63 +/- 0.04 for CCP and Y-516, respectively. Y-516 was detected in the plasma after intraperitoneal and oral administration of CCP. The ratio of the AUC of Y 516 to that of CCP was 0.06 following intraperitoneal administration and 0.40 following oral administration. This indicated that while the metabolism of CCP into Y-516 may occur partly in the liver due to the first-pass effect, it occurs mostly within the gastrointestinal tract itself or its mucosa.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566186 TI - Behavioural effects of the benzodiazepine receptor partial agonist RO 16-6028 in mice. AB - The imidazo-diazepinone RO 16-6028 is a benzodiazepine receptor partial agonist which exhibits some anti-conflict effects in the two-chambered light/dark test without significantly affecting the behaviour of mice confronted with the staircase test. In addition, this drug slightly reduced locomotion and more markedly rearing in a free exploration procedure. These results indicate that RO 16-6028 appears to produce some anxiolytic and sedative properties like full agonists, but with weaker magnitude. This could be related to the benzodiazepine partial agonistic profile of the compound. PMID- 2566188 TI - Lack of dyskinesias in unmedicated schizophrenics. PMID- 2566187 TI - Attenuation of defensive threat and attack in wild rats (Rattus rattus) by benzodiazepines. AB - A battery of tests designed to elicit reactions to a variety of non-painful threat stimuli was used to study the effects of chlordiazepoxide (5-20 mg/kg), diazepam (1-5 mg/kg) and midazolam (1-10 mg/kg) on the defensive repertoire of wild Rattus rattus. The most consistent effect of benzodiazepine treatment, across compounds and tests, was a marked reduction in defensive threat and attack behaviors, with midazolam effective over a wider range of situations. In contrast, effects on freezing and flight reactions were more variable, differing substantially as a function of stimulus context. The general profile of observed changes in defense cannot be explained in terms of either non-specific behavioral suppression or a global reduction in defensiveness. Rather, our findings suggest that benzodiazepines may primarily induce a shift within the defense repertoire. PMID- 2566189 TI - In search of cellular mechanisms of memory. PMID- 2566190 TI - Neuropeptides in Alzheimer's disease: a postmortem study. AB - The concentration of 5 neuropeptides, neurotensin (NT), somatostatin (SRIF), corticotropin-releasing factor (CRF), bombesin and thyrotropin-releasing hormone (TRH) was measured in 3 cerebrocortical areas and several subcortical regions in post-mortem brains obtained from patients with histologically verified Alzheimer's disease and from controls without neurological or psychiatric disorders using sensitive and specific radioimmunoassay procedures. In Alzheimer's disease, reductions in the concentration of SRIF and CRF were observed in frontal and temporal cortex. In addition, in Alzheimer's disease, SRIF was also reduced in concentration in the hypothalamus, whereas CRF concentrations were reduced in the caudate nucleus. Neurotensin was reduced in concentration in the amygdala in Alzheimer's disease. No alterations in TRH or bombesin/gastrin-releasing peptide were found. These findings provide further evidence for the pathological involvement of certain neuropeptide-containing neurons in Alzheimer's disease. PMID- 2566191 TI - Antisecretory effects of somatostatin and vasopressin in the rat colon descendens in vitro. AB - The effects of two hormones, vasopressin and somatostatin (SOM), on ion secretion in rat colon descendens were compared. Three modes for induction of epithelial secretion were used: neuronally mediated secretion due to electric field stimulation (EFS), Ca2+-dependent secretion elicited by carbachol, and cAMP dependent secretion evoked either by a receptor-mediated mechanism elicited by vasoactive intestinal peptide (VIP) or by a direct activation of the adenylate cyclase by means of forskolin. Somatostatin inhibited ion secretion evoked by EFS (55-65%), carbachol (80%) and VIP (95%) in a dose-dependent manner. Maximal inhibition by SOM was observed at 10(-7) M. Somatostatin had, however, no effect on the secretory response to forskolin. The inhibition of the VIP effect could be attenuated by pretreatment with pertussis toxin. In contrast, vasopressin in concentrations as low as 0.025-0.25 U/liter decreased the secretory effects of EFS (55-75%) and carbachol (85%), but had no effect on cAMP-dependent secretion elicited either by VIP or forskolin. The results suggest that the antisecretory effect of vasopressin is mediated only by a block in the Ca2+ pathway, whereas SOM inhibits Ca2+-dependent secretion as well as receptor-mediated cAMP-dependent secretion. The interaction with the cAMP pathway is located at the step between stimulation of the receptor and activation of the adenylate cyclase and probably involves an Ni-protein. PMID- 2566192 TI - Studies on the inspiratory generating effect of the dorso-medial area of nucleus facialis. AB - The responses in phrenic nerve activity resulting from stimulation and successive focal block of the dorso-medial area of nucleus facialis (dmNF) were studied in urethane anaesthetized rabbits. It was found that: (1) Long train stimulation (0.025-0.2 mA, 100 Hz) delivered to the dmNF caused marked prolongation in inspiratory duration and distinct increase in amplitude of phrenic discharge. Continuous inspiration with increased amplitude was usually observed during stimulation. (2) Short train stimulation (10 pulses, 100 Hz, 0.025-0.2 mA) during inspiration increased the duration and amplitude of inspiration. When the short train stimulation was delivered during expiration, inspiration would be initiated prematurely. (3) Microinjection of L-glutamate (0.5 M, 0.5-2 microliters) into the dmNF also increased inspiratory duration and amplitude. (4) Focal block of the dmNF by microinjection of lidocaine (2%, 1-2 microliters) produced strong depression or complete cessation of inspiration. In conclusion, the results of the present study show that the dmNF may play an important role in the neurogenesis of respiratory rhythm. PMID- 2566193 TI - [Systemic complications of beta-blocking eyedrops. Apropos of 6 cases]. AB - Six cases of systemic reactions to topical treatment with beta-blocking eyedrops are reported, bradycardia and faintness due to an overdosage of ophthalmic timolol; decompensated heart failure one month after the prescription of carteolol eyedrops: bronchospasm after two weeks of treatment with metipranolol eyedrops; crippling Raynaud's phenomenon of otherwise unknown origin, which had begun with timolol eyedrops, continued with carteolol eyedrops and regressed after discontinuation of ophthalmic beta-blockers; aggravation of an anaphylactoid shock in a patient treated with ophthalmic timolol, and myocardial infarction possibly due to the abrupt withdrawal of timolol eyedrops. It cannot be overstressed that the rules governing the prescription of oral beta-blockers also apply to ophthalmic preparations of these drugs: respect of contra indications, strict adherence to the dosage recommended, gradual drug withdrawal and regular supervision. Only controlled studies and long-term follow-up will be able to demonstrate differences in safety between the five beta-blockers commercialized as eyedrops in this country. PMID- 2566194 TI - [An autopsy case of malignant histiocytosis-like disorder following hypersensitive reaction to mosquito bite]. AB - An autopsy case of an 18-year-old Japanese girl with a malignant histiocytosis like disorder that developed during the course of a hypersensitive reaction to mosquito bite is reported. Episodes of hypersensitive reactions to mosquito bite had been repeated since she was 12 years old and at the age of 18 years she died of acute respiratory failure only 11 days after a mosquito bite. On autopsy, the dermal reaction to the last mosquito bite had already calmed down, and 'poorly differentiated histiocytes', which were presumed from their histochemical characteristics, had remarkably infiltrated into multiple organs, appearing like leukemic infiltration. Only a small number of them were noted in the lymph nodes and the bone marrow. The results of histochemical examination of 'poorly differentiated histiocytes' was as follows: (1) Neither markers for granulocytes (peroxidase and naphthyl AS-D Cl esterase) nor markers for lymphocytes and plasma cells (leukocyte common antigen and immunoglobulins) were detected. (2) Some markers for histiocytes (peanut lectin agglutinin and lysozyme) were positive in some of the proliferated cells. (3) A marker for T-zone histiocyte (s-100 protein) was negative. These results suggested that the proliferated cells included cells of the monocyte-macrophage system. These cells morphologically showed no phagocytic activity and were suggested to be immature histiocytes. Jurco et al. reported (poorly differentiated) malignant histiocytosis consisting of immature histiocytes without phagocytic activity, by using histochemical methods.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566195 TI - [The treatment of convulsions in children]. PMID- 2566196 TI - [Optimal use of antihistaminics]. PMID- 2566197 TI - [The correct use of beta 2 stimulants]. PMID- 2566198 TI - Intermittent medication for schizophrenic outpatients: who is eligible? AB - The Medication Clinic of a large, urban Mental Health Center was screened for schizophrenic patients eligible for an intermittent medication approach. A total of 112 patients were evaluated, and 39, or 34.8 percent of the sample, met our basic inclusion criteria. No sex or age differences were found for eligibility. Sufficient eligible patients were found to make the intermittent medication approach a useful part of a comprehensive psychopharmacological program for schizophrenia, if the efficacy of the approach is demonstrated in clinical trials. PMID- 2566199 TI - Response to neuroleptic drugs as a device for classifying schizophrenia. AB - Although schizophrenic patients are routinely treated with neuroleptic medication, the diversity in response to such treatment is noteworthy; some patients are exquisitely responsive to neuroleptic treatment, while others are clearly resistant. The authors examine the hypothesis that neuroleptic-responsive and neuroleptic-resistant patients have different illnesses by considering the following issues: the reliability of the distinction between neuroleptic responsiveness and resistance; the consistency in neuroleptic responsiveness over time; the association between neuroleptic responsiveness and other clinical features; and the neuroleptics' therapeutic action. On the basis of the data available and on theoretical and historical grounds, the distinction between neuroleptic-responsive and neuroleptic-resistant patients warrants application in both clinical and research settings. PMID- 2566201 TI - Two cultures find common ground. PMID- 2566200 TI - [Stress ulcer disease and its prevention]. AB - The incidence and prophylaxis of stress ulcer disease are analyzed. Acute gastroduodenal stress lesions are frequently encountered in critically ill patients, yet the incidence of stress ulcer bleeding has decreased. It is still difficult to describe the risk in an individual patient in spite of the fact that patient groups prone to development of stress ulcer disease have been well defined (e.g. patients on mechanical ventilation). Prophylaxis with antacids, H2 receptor antagonist, pirenzepine and sucralfate seems to be equally effective and capable of reducing the incidence of stress ulcer bleeding. Its efficacy does not really correlate with the capability of a drug to keep gastric pH above 4. The higher the incidence of stress ulcer bleeding under placebo, the better the efficacy of prophylactic treatment. There is evidence that blockers of gastric acid secretion may be associated with an increased incidence of nosocomial pneumonia, but more data are needed in this regard. PMID- 2566202 TI - Gene signals relapse of breast, ovarian cancers. PMID- 2566203 TI - Intracranial complications of sinusitis. AB - Sinusitis is a common problem that is routinely diagnosed and treated by most primary care physicians. Although most cases usually respond to appropriate therapy, some occasionally progress to the development of intracranial complications, including meningitis, osteomyelitis, epidural and subdural empyema, intracranial mucocele or polyps, and frank brain abscess. It is important to develop a rational approach to the diagnosis and treatment of these conditions. A high clinical index of suspicion must always be maintained, since symptoms are often masked by previous antibiotic therapy. Radiologic evaluation must always include computerized tomography (CT) for accurate diagnosis and surgical planning. Therapy includes surgical drainage and high doses of appropriate intravenous antibiotics. Cefuroxime and metronidazole provide excellent broad spectrum antibacterial coverage. Only early recognition and appropriate therapy can reduce the potential morbidity and mortality associated with these life-threatening complications. PMID- 2566204 TI - Weight gain associated with psychotropic drugs. AB - The weight increase that is an often reported side effect of psychotropic drug use has implications for health risks as well as medication noncompliance. In this article I review the literature on the association between the use of several classes of psychotropic drugs and weight gain, discuss the possible mechanisms for this effect, and offer some practical approaches for reducing the risk of drug-induced weight gain, including the identification of a group of patients "at risk." PMID- 2566205 TI - In vivo quantification of blood-brain transfer and binding of [125I]HEAT, an alpha 1-adrenoceptor antagonist. AB - The uptake and binding constants of [125I]iodo-2-[beta-(4-hydroxyphenyl)-ethyl amino-methyl]tetralone ( [125I]HEAT) in rat brain were determined in vivo. The initial clearance of the radioligand from blood to brain, K1, was calculated from the initial uptake of the radioligand; it averaged 0.21 +/- 0.01 (SD) ml g-1 min 1, consistent with an initial extraction of 25% (i.e., one-quarter of the blood flow). The most strongly binding regions included the olfactory bulb, thalamic nuclei, medial geniculate body, and cerebral cortical layers. We identified saturable, specific binding in frontal cortex layers 1, 5a, and 5c (motor region), frontal cortex layers 3+4, ventral thalamic nuclei, medial geniculate body, striatum, cerebellum, and olfactory bulb. Addition of unlabeled ligand depressed binding in all regions to the same low level (partition coefficient) of 0.8 ml g-1. Displacement of [125 I]HEAT binding by unlabeled HEAT yielded a global affinity constant (KDVd) of 34 +/- 8 pmol g-1 and receptor densities (Bmax) that varied from 50 pmol g-1 in cerebellar cortex and caudate nucleus to 200 pmol g-1 in the region of highest specific binding, the medial geniculate body. PMID- 2566207 TI - Effects of toxin isolated from the sea anemone Bolocera tuediae on electrical properties of isolated rat skeletal muscle and cultured myotubes. AB - Electrophysiological effects of a polypeptide toxin isolated from the sea anemone Bolocera tuediae (BTTX II) on isolated fast and slow rat skeletal muscle and on cultured rat myotubes are described in this paper. Nanomolar concentrations of BTTX II cause a concentration-dependent depolarization. This effect is prevented by the presence of tetrodotoxin. Action potential duration is prolonged in all preparations investigated, with the fast muscle being the least sensitive. The positive overshoot and the maximum rate of rise of the action potential is decreased by BTTX II. Cultured myotubes which are less susceptible to the sodium channel blocking activity of tetrodotoxin have the highest sensitivity to all actions of BTTX II. It is further shown that the proportion of spontaneously contracting myotubes is diminished and the frequency of contractions is increased by the toxin. The effects of Bolocera tuediae toxin II resemble those observed with Anemonia sulcata toxin II on mammalian skeletal muscle. From the results it is concluded, that BTTX II may interfere with both the activation and inactivation of sodium channels of the muscle membrane. PMID- 2566206 TI - [The determination of blood parameters by Reflotron]. AB - In this paper, the practicability of a quick diagnostic laboratory test to perform blood analysis in healthy pigeons was tested. The Reflotron manufactured by Boehringer Mannheim, works on a dry chemical basis, consists of a reflectance photometer and the accompanying test strips, is easy to use, and yields the result of the desired parameter in a maximum of three minutes. As test material plasma and serum were used. At the time the equipment was tested, test strips for glucose, cholesterol, hemoglobin, triglycerides, urea, uric acid, GOT, GPT and G GT were available on the market. The usual laboratory "wet chemical" analysis procedures were used to compare the results. The obtained measurements were analyzed using describing and concluding statistics. The influence of gender, feed and time of day on the nine parameters was also considered. PMID- 2566208 TI - First assisting in cataract surgery: program results. AB - 1. This program was established to teach non-physicians to first assist in cataract surgery, since third party payors would no longer pay for physicians to first assist. 2. Among the advantages to utilizing non-physician first assistants are: availability, reliability, familiarity with instruments and equipment, and quality of service. 3. The questionnaire results confirm that the advantages of having non-physician first assistants far outweigh the disadvantages. PMID- 2566209 TI - Peripheral blood stem cell collection and use in Hodgkin's disease. Comparison with marrow in autologous transplantation. AB - Hematopoietic progenitor cells can be collected from blood by cytapheresis; the clinical use of these cells may offer such advantages over marrow as the avoidance of general anesthesia, collection on an outpatient basis, and use when marrow is involved with malignancy. Since Hodgkin's disease rarely spreads hematogenously, postchemotherapy marrow transplantation with autologous peripheral blood stem cells (PBSCs) was compared to that with marrow transplantation in patients with this disorder. Seven patients were treated with PBSCs and 19 with marrow. Five to nine collections of PBSC were performed per patient. There was a rebound increase in circulating committed progenitors when PBSC were collected during the marrow rebound after cyclic chemotherapy. After intensification and cellular rescue, quicker recovery of circulating white cells (p less than 0.05) and a shorter hospital stay (not significant) were seen in the PBSC patients than in those treated with autologous marrow. There was no difference in the duration of red cell or platelet transfusion required after transplant. Of six patients whose marrows were previously involved by Hodgkin's, recurrent or progressive disease has occurred in five. PBSC may be a viable alternative to marrow in selected patients. PMID- 2566210 TI - [Ulcer surgery on the cross road?]. PMID- 2566211 TI - [Functioning of the artificial bladder after fistuloplasty by the transvaginal approach]. PMID- 2566212 TI - Linkage of the gene for the scrapie-associated fibril protein (PrP) to the Sip gene in Cheviot sheep. AB - The gene Sip with two alleles, sA and pA, is the major gene determining the incubation period of scrapie in its natural host, sheep. Two lines of Cheviot sheep have been bred which differ in their response to experimental infection with SSBP/1 scrapie. The negative line have a decreased incidence of disease caused by SSBP/1 and are SippApA. The positive line have an increased incidence of disease and the majority are either SipsAsA or SipsApA; it is not possible to distinguish between the two genotypes on the basis of scrapie incubation time because the sA allele is fully dominant with SSBP/1 scrapie. There are also rare SippApA segregants in the positive line. The major protein (PrP) of scrapie associated fibrils is encoded by a cellular gene and a cDNA copy of the hamster PrP mRNA has been used to analyse the restriction fragment length polymorphism of the two lines of Cheviot sheep. Two polymorphisms of the sheep PrP gene were found, by using HindIII and EcoRI, which appear to act as markers for the alleles of Sip. Using these polymorphisms it is now possible to assign a Sip genotype to the sheep in the Cheviot flock. Preliminary results from Anglo-Nubian goats and a cow are also reported. PMID- 2566213 TI - [Lymphocyte subpopulations in patients with skin basalioma of the trunk]. AB - In 11 male patients with histologically proven basal cell epithelioma of the trunk, we studied the distribution of peripheral lymphocyte subsets. All of the patients revealed pathological T-cell subsets, 6 of them showing reduced numbers of CD8 and 5 decreased numbers of CD 4 cells. 2 patients had taken arsenic; in 2 patients we diagnosed a further tumor of some other origin. PMID- 2566214 TI - [Aspects of genetic technics in diabetes research]. PMID- 2566215 TI - [Injuries and damage caused by excessive stress in classical ballet]. AB - Typical injuries of highly professional classical ballet dancers were evaluated in 108 members of German opera companies. We demonstrated the predominance of overuse-syndromes to acute injuries related to the excessive training-extends. Disorders of the feet, particularly tendinitis followed by low-back pain, chondromalacia patellae, jumpers knee, shin splints and muscle strains are most frequent. Considerable time often elapsed before diagnosing was made because complaints were ignored by dancers. Our inquiry includes range of motion of the spine and the extremities, serial x-rays and a systematic review of ballet related injuries, training-programs and techniques. PMID- 2566216 TI - [Pharmacologic aspects of the treatment of drug addiction due to morphine-like substances]. PMID- 2566217 TI - [Side effects of psychopharmacotherapy in patients with schizophrenia proceeding against a background of organic cerebral insufficiency]. AB - Analyzed were 1999 courses of treatment in 212 patients with paroxysmal and paranoid schizophrenia. Twelve groups of side-effects were defined according to syndromal and structural principles. Compared to the uncomplicated schizophrenic patients, the ones with organic cerebral insufficiency had higher sensitivity to psychopharmacologic drug therapy. The number of untoward effects per one course of treatment in organically deficient patients was 1.5 times higher than that of the control group. It was also shown that cerebral organic insufficiency promoted the long-lasting and chronic extrapyramidal disorders. It also increased the rate of akathisia, akinetic-rigid syndrome, hyperkineses, fever, epileptiform paroxysms, circulatory-autonomic disorders. Side-effects were closely related to the nature of drug therapy. PMID- 2566218 TI - [Amino acid reserves of the blood in patients with schizophrenia and their correction in insulin coma]. AB - Free amino acid content was measured in the blood serum in patients with permanently progressive paranoid schizophrenia under conventional insulin shock therapy. The influence of several amino acids (methionine, glutamic acid) preinjection on their base levels was also assessed. Metabolic response to the insulin coma was different in preinjected vs. control groups. Glutamine acid preinjected patients displayed less pronounced shifts in amino acid reserves in the blood serum that smoothed the shock-related metabolic stress in these patients as related to conventional shock. PMID- 2566219 TI - Some aspects of the experimental induction and measurement of itch. AB - Two different rating scales--a visual analogue scale (VAS) connected to a chart recorder, and Pain-Track, a micro-computerized system with a 7-step-graded, fixed point, non-verbal scale (FPNVS)--were evaluated for their capacity to assess experimental, histamine-induced itch continuously in 38 healthy subjects. The consequences for itch perception of using different injection sequences of various histamine concentrations were also investigated. A linear dose-response relationship was shown with random injection order for all subjective variables studied (itch latency and duration, maximal itch intensity, 'total itch index') with the VAS, but only for itch duration and 'total itch index' with the FPNVS. Using the VAS and injecting histamine solutions with increasing concentration, a significant dose-response curve was obtained for maximal itch intensity and 'total itch index', but when the same histamine stimuli were presented in the reverse (i.e. decreasing) order, there was no dose-response relationship. This indicates that central nervous system interaction may be unequally activated, depending on the order of different injected histamine stimuli. The objective variable flare was unaffected by the injection sequence. It is concluded that random injection order should be used in the assessment of itch sensation, in order to avoid systematic errors. The fact that the FPNVS did not discriminate as well as the VAS could indicate that our experimental stimuli were too weak to be properly discriminated with a 7-step-graded scale. PMID- 2566220 TI - An experimental study evaluating the effect of minoxidil on the growth cycle of hair follicles. AB - The possibility that topically-applied minoxidil might affect the growth cycle of hair follicles was studied in inbred Herston white mice and HRA/Skh1 hairless mice. In the normal follicular cycle, the anagen or growth phase can be followed by autoradiographic demonstration of [3H]thymidine uptake in proliferating matrical cells, and the catagen or regression phase can be recognised, using light microscopy, by the presence of greatly increased death of matrical cells by apoptosis. Using these two markers, the effects of topically-applied minoxidil on follicular kinetics were studied, during neonatal hair growth and the spontaneous wave of hair loss that occurs 16 to 17 days after birth. Minoxidil at strengths of either 1% or 3%, applied daily to the dorsal skin of newborn mice from birth until 25 days of age, was found to have no recognisable effect. Despite this negative result, however, the study does show the potential for the use of apoptosis as a marker for catagen in research in dermatopathology. PMID- 2566221 TI - Interleukin-6 in the epidermis of patients with psoriasis before and during PUVA treatment. AB - Biopsies from lesional and unaffected skin of 6 patients with psoriasis, taken before and during treatment with psoralen plus UVA (PUVA) were examined immunohistologically, using partially purified polyclonal antibodies to crude supernatants of activated human blood monocytes. By absorption with recombinant derived human monokines, we were able to demonstrate that interleukin-6 (IL-6) (but not IL-1 alpha or IL-1 beta) was located in a laminar and granular pattern in stratum corneum, and on epidermal cell membranes in the viable cellular epidermis. Before PUVA treatment, the intensity and the extension of staining for IL-6 were both markedly increased in lesional skin compared with uninvolved skin. A weaker staining for IL-6 was observed in lesional skin, simultaneous with the clinical improvement of psoriasis. The staining patterns for IL-6 in biopsies from cleared lesional skin and uninvolved psoriatic skin were identical at the conclusion of therapy. PMID- 2566222 TI - Epidermal Langerhans' cells in chronic eczematous dermatitis of the palms treated with PUVA and UVB. AB - Epidermal Langerhans' cells (LC) were studied in patients with chronic eczematous dermatitis of the palms. The monoclonal antibodies anti-Leu 6 and anti-HLA-DR were used, and the cells visualized with an immunoperoxidase technique. Increased numbers of LC were found in allergic contact dermatitis as well as in irritant contact dermatitis and hyperkeratotic dermatitis of the palms. The increased number of epidermal LC in lesional skin may facilitate presentation of exogenous or endogenous antigens to activated T-cells. One hand was treated with PUVA or UVB and the other hand served as a non-exposed control. PUVA treatment cleared the dermatitis and the LC number decreased markedly. With UVB treatment clinical improvement was achieved, and a less pronounced decrease in epidermal LC was noticed. There seems to be a crude relationship between the extent of clinical improvement and the reduction in epidermal LC numbers. The change in LC might be a primary event or secondary to a reduction of the inflammatory process. PMID- 2566223 TI - The development of a Ki-1-positive large cell non-Hodgkin's lymphoma in pagetoid reticulosis. AB - A case of the disseminated variant of pagetoid reticulosis is described which progressed after many years of disease to a large cell anaplastic (Ki-1) lymphoma of T-cell type. The lymphoma cells showed abnormal cellular DNA and a high proliferative rate, as revealed by immunophenotypic examination and single-cell DNA measurements. The cells were positive for activation associated antigens and expressed a T-helper/inducer phenotype. A similar phenotype was expressed by the neoplastic cells in the co-existing lesions of pagetoid reticulosis. These findings support the view that pagetoid reticulosis is a variant of the cutaneous T-cell lymphomas which originates from activated lymphoid cells and which may on occasion progress to a potentionally more aggressive lymphoid malignancy. PMID- 2566224 TI - Hyperpigmented acral papular mucinosis, systemic lupus erythematosus and universal alopecia. AB - A 42-year-old man presented with systemic lupus erythematosus, universal alopecia and non-pruritic hyperpigmented papular mucinosis. The latter was most evident on acral areas. In hyperpigmented areas of the face the immunofluorescence showed deposits as in LE and with alcian blue and colloidal iron an abundance of mucin was demonstrated in the dermis. A lesion on the back showed only papular mucinosis. Fifteen cases of LE and papular mucinosis reported in the literature are reviewed. Our patient differs with respect to the marked pigmentation of his lesions, their localization and the association with universal alopecia. PMID- 2566226 TI - Epidemiology of different types of hand eczema in an industrial city. AB - Different types of hand eczema in an industrial city were studied. Questionnaires were sent to 20,000 individuals aged 20-65 years, randomly selected from the population register of the city. Those subjects (1,385) considering themselves to have had hand eczema within the previous 12 months were invited to a dermatological examination. It was found that hand eczema occurred twice as often among females as among males. The most common diagnosis was irritant dermatitis. Atopic hand eczema and allergic contact dermatitis had a lower but approximately equal prevalence. Onset of hand eczema at young ages was common, in particular among women. Hand eczema was shown often to be a long-lasting disease with a relapsing course. Atopic hand eczema seemed to be most unfavourable, with a long duration, high continuity of symptoms and extensive involvement. PMID- 2566225 TI - Cutaneous drug reactions: clinical types and causative agents. A five-year survey of in-patients (1981-1985). AB - We collected a 5-year series of drug eruptions. There were 225 cases, 128 of them verified by a positive provocation test. The most common types of clinical reaction were fixed drug eruptions, exanthematous eruptions and urticarias. The drugs most often responsible for the eruptions were antimicrobial agents and antipyretic/anti-inflammatory analgesics. Comparing this series with our three previous series from the same hospital, the total number of drug eruptions proved to have decreased over the last 30 years. The main groups of drugs causing skin reactions have remained the same, but in recent years the proportion of sulphonamides has diminished. PMID- 2566227 TI - Racial differences in mole proneness. AB - Mole counts were studied in relation to skin complexion in various racial groups. White children had a median total number of naevocytic naevi of 17.0, versus 2.5 in non-white children (p less than 0.001). Young white adults showed a similar mole proneness to that of coloured subjects (61.0 versus 16.0; p less than 0.001). With regard to moles greater than 2 mm diameter in the young-adult group, white subjects again exhibited a higher median count than non-white subjects (5.5 versus 1.0; p less than 0.001). There was an inverse gradient of mole counts in young adults from subjects of white complexion through those of mixed ancestry, Oriental ancestry, to those of Negroid descent. This study indicates that there is a strong racial background predisposing to the development of naevocytic naevi. PMID- 2566228 TI - Irritancy of dithranol in normally pigmented and depigmented skin of patients with vitiligo. AB - In order to ascertain the extent to which the pigmentary system plays a protective role in dithranol-induced irritancy, a within-subject comparison was carried out between normally pigmented and depigmented skin of patients with vitiligo. In open patch tests, various concentrations of dithranol in a cream base were applied to the normally pigmented and depigmented skin of 6 patients with vitiligo. The responses were assessed 48 h after application. A mild to moderate inflammation occurred in the pigmented and depigmented skin and no statistically significant difference was shown between the two test areas. The present study does not support the hypothesis that the pigmentary system might be involved in dithranol-induced irritancy. PMID- 2566229 TI - Melanocytic proliferation in condylomata acuminata. A report of two cases and investigation by in situ hybridisation. AB - Two cases of melanocytic lesion occurring in condylomata acuminata are described. In situ hybridization with human papillomavirus (HPV) DNA probes specific for 6b, 11, 16, 18 revealed positivity with HPV6 and 11, in the non-dysplastic condylomata, and HPV 18 positivity in the case showing severe dysplasia. The HPV localization was confined to the superficial parakeratotic zones, remote from the melanocytic proliferation. The relationship between human papillomavirus and the melanocytic lesions is discussed. PMID- 2566230 TI - Serum aminoterminal propeptide of type III procollagen. A non-invasive test for liver fibrogenesis in methotrexate-treated psoriatics. AB - Serum aminoterminal propeptide of type III procollagen (PIIINP) was studied in 73 psoriatics receiving methotrexate and in 11 selected for trial with methotrexate or etretinate. 72 of the patients on methotrexate were also investigated with liver biopsies. The highest PIIINP value was found in a patient with ascites and her PIIINP decreased after medication was discontinued. Psoriatics with fibrosis or cirrhosis in their liver biopsies had a significantly higher mean PIIINP than patients without fibrosis, who had the same mean value as psoriatics prior to treatment. Based upon the individual data together with data from serial PIIINP investigations of 11 patients studied during treatment, it is concluded that PIIINP can be utilized as a valuable non-invasive test for liver fibrogenesis in methotrexate-treated psoriatics. PIIINP is not specific for the liver, but the study indicates that the number of liver biopsies can be reduced in psoriatics on methotrexate who have normal levels of PIIINP. PMID- 2566232 TI - Experimental dermatophyte infection. The extent of the fungal invasion. AB - A spore suspension from a granular strain of Trichophyton mentagrophytes under occlusion for 4 days was used to produce two fungal lesions on the upper arm of the Trichophytin-negative author. The material for culture was obtained by stripping across the visible lesions and several centimetres into the surrounding skin. Already when the occlusion was removed, the whole stratum corneum was heavily invaded up to 20 mm into the surrounding, normal-looking skin. When the intensity and the size of the visible lesions topped after about 2 weeks, the culture positivity reached 45 mm into the perilesional skin. One week later the spontaneous involution had eliminated nearly all fungal organisms through the entire horny layer of both the visible lesions and their surroundings. After 42 days the lesions were culture-negative. It was concluded that the growth pattern of an experimental infection could be as observed in natural infections. PMID- 2566231 TI - Trichophyton rubrum abscesses in immunocompromised patients. A case report. AB - A 72-year-old immunocompromised man with myelodysplastic syndrome who developed multiple erythematous, scaly abscesses like lesions on his left foot and lower leg is described. He also had dry scaly lesions on his soles and lesions on several toe nails. A punch biopsy showed abscesses with fungal elements and Trichophyton rubrum was cultured from skin scales and the biopsy. A diagnosis of T. rubrum abscesses should be suspected in all immunocompromised patients with signs of superficial dermatophyte infection. PMID- 2566233 TI - Cyclosporin A does not modify Langerhans' cell number and distribution in normal human skin. AB - We used the model of human skin graft on nude mouse to investigate a possible influence of systemically administered cyclosporin A (CsA) on epidermal Langerhans' cells (LC). This experimental model has the advantage of being independent of the systemic humoral and cellular influences of a human host. No change in the human LC distribution or number could be observed after 3 weeks of CsA therapy as revealed by anti-CD1 and anti-HLA-DR immunohistochemical stainings and the cell counting, despite the evidence of the drug effect on the keratinocyte proliferation. However, our findings do not rule out the possibility that CsA influences the LC functional capacities responsible for the local cell mediated immunity responses. PMID- 2566234 TI - Topical cyclosporin A in alopecia areata. AB - We conducted a trial of topical application of 10% cyclosporin A in an oil preparation in 10 patients with alopecia areata and alopecia universalis. After 12 months of therapy, no beneficial response was observed in any of the 10 patients. PMID- 2566235 TI - Prostaglandin E1 and prostaglandin F2 alpha in exudate in nickel allergy. AB - Ten nickel-allergic patients and 5 healthy control subjects participated in a study of the kinetics of the flux and concentration of migrated leukocytes and extracellular PGE1 and PGF2 alpha during a 48 h period, using a skin chamber technique. The patients were provided with two skin chambers, one with and one without nickel challenge. A higher flux of leukocytes, PGE1 and PGF2 alpha was observed during the second day of allergen exposure, while the concentrations probably due to dilution were unchanged or diminished, indicating an unspecific role of the prostaglandins during the contact allergic reaction. No correlations were found within the groups between the migration of leukocytes and the prostaglandin content. PMID- 2566236 TI - Unilateral eruption of endogenous eczema after hemiparesis. AB - Five patients with cerebrovascular hemiplegia developed an endogenous eczema (nummular eczema, pompholyx, allergids, atopic dermatitis). In all cases the dermatitis was mainly confined to the healthy side. PMID- 2566237 TI - Unusual localization of lichen amyloidosus. Topical treatment with dimethylsulfoxide. AB - An unusual localization of lichen amyloidosus in a patient with IgG k benign monoclonal gammopathy is reported. After topical treatment with dimethylsulfoxide the lesions improved, but histological examination still showed amyloid deposits. PMID- 2566238 TI - A survey of elderly new patients at a dermatology outpatient clinic. AB - A study of all new dermatitic out patients greater than 70 years of age, referred in the period January-June 1987 (257 patients), was undertaken to obtain information on the types of skin diseases and the dermatological outpatient services required in the case of the elderly. The group accounted for 14% of all new patients. The leading diagnoses were seborrheic keratosis (15.6%), basal cell carcinoma (13.6%), solar keratosis (13.2%), psoriasis (9.7%) and leg ulcer (9.3%). A skin biopsy was required in 27%, and surgical treatment in 31% of the group, compared with 12% and 16%, respectively, of all new patients during the same period. Consequently, the elderly group were more likely to require a follow up appointment and tended to be more time-consuming outpatients. PMID- 2566239 TI - Transformation of myelodysplasia to acute myeloid leukaemia during psoralen photochemotherapy (PUVA) treatment of psoriasis. AB - A patient with stable chronic myelomonocytic leukaemia was treated with 8 methoxypsoralen photochemotherapy (PUVA) for erythrodermic psoriasis. After 4 months, transformation to acute myeloid leukaemia (AML) occurred. We would suggest caution when considering PUVA therapy in patients with pre-leukaemic disorders. PMID- 2566240 TI - Monoamine- and diamine oxidase activities in psoriasis. AB - Monoamine- and diamine oxidase activities were measured by a sensitive photometric assay in 25 psoriasis vulgaris patients. Results were compared with plasma histamine values determined fluorimetrically. Increased plasma histamine levels were associated with significantly lowered diamine--and type B monoamine oxidase activities in platelet-rich plasma of the psoriasis patients. Our data suggest that cofactor levels and/or inhibiting factors are responsible for the observed monoamine- and diamine oxidase activities. PMID- 2566241 TI - Dietary supplementation with a combination of n-3 and n-6 fatty acids (super gamma-oil marine) improves psoriasis. AB - Psoriasis may improve during dietary supplementation with fish oil containing n-3 fatty acids including eicosapentaenoic acid. In the present study 17 psoriatic patients were treated with Super Gamma-Oil Marine containing a combination of n-3 and n-6 fatty acids (linoleic acid and gammalinolenic acid). After 4 months, excellent improvement was observed in 2 patients, moderate improvement in 8, mild improvement in 4, and no improvement in 3 patients. These results may indicate that a combination of n-3 and n-6 fatty acids is useful for the treatment of psoriasis. However, controlled studies including more patients are warranted. PMID- 2566242 TI - Multiple bites from the larvae of the tick Ixodes ricinus. A case report. AB - The case is presented of a 32-year-old man with multiple reddish-blue papules on both thighs and sporadic on the lower part of the trunk and lower legs, clinically compatible with multiple 'insect' bites. The lesions were found to be multiple bites from larvae of the tick Ixodes ricinus. The larvae are about 1 mm long, live very close to the ground, and mainly parasitize small rodents. They are, therefore, generally not well known. Although seldom encountered, we suggest that bites from the larvae of the tick Ixodes ricinus should be kept in mind in patients with multiple 'insect' bites, especially in periods with high tick activity. The differential diagnosis is important because the larvae may cause infection with the spirochete Borrelia burgdorgferi. PMID- 2566243 TI - Start and end of the effects of terfenadine and astemizole on histamine-induced wheals in human skin. AB - A study was made of effects of two antihistamines, terfenadine (60 mg twice daily) and astemizole (10 mg once daily) on wheals induced by histamine dihydrochloride (10 mg/ml) in the prick test on the upper back of 15 healthy students. The suppressive effects of terfenadine on the histamine wheal appeared earlier (2 h), and disappeared earlier (within 1 day) than those of astemizole (3 days and 28 days, respectively). No difference between the maximal effects of the two drugs was seen. PMID- 2566244 TI - Lack of effect of cyclosporin A in pityriasis rubra pilaris. PMID- 2566245 TI - Erythroderma, seborrheic keratoses and Leser-Trelat sign. PMID- 2566246 TI - Development of metastatic skin cancer during methotrexate therapy for psoriasis. PMID- 2566247 TI - Graves' hyperthyroidism following primary hypothyroidism: sequential changes in various activities of thyrotropin receptor antibodies. AB - A 40-year-old male who developed Graves' hyperthyroidism following primary hypothyroidism is reported. He presented with clinical signs of hypothyroidism and concomitant myasthenia gravis. The thyroid was not palpable. He was treated with T4, pyridostigmine and prednisolone. One year later he developed hyperthyroidism and goitre. His initial serum IgG had no intrinsic thyroid stimulating activity, but showed almost complete inhibition of TSH-stimulated cAMP generation (99.4%, normal less than 38%) and [3H]thymidine incorporation (99.5%, normal less than 40%) into rat thyroid cells, FRTL-5 cells, with very high activity (80.2%, normal less than 15%) of TSH binding inhibitor immunoglobulin. When he developed hyperthyroidism and goitre, his IgG showed a strong thyroid stimulation, both cAMP production (27-fold increase) and [3H]thymidine incorporation (5.5-fold increase). No inhibitory activities were noted. These findings suggest that clinical states of autoimmune thyroid diseases can be changed in accordance with changes of functional properties of TSH receptor antibodies. PMID- 2566248 TI - Clobazam increases pulsatile luteinizing hormone secretion in normal men. AB - To investigate the effects of the 1,5-benzodiazepine, clobazam, on LH secretion in normal men, LH pulsatile secretion was defined after oral administration of 40 mg of clobazam or a placebo to 6 healthy male volunteers, according to a randomized cross-over design. LH pulse frequency increased significantly from a mean of 3.8 (range 3-5) pulses/8 h after placebo, to a mean of 5 (range 4-7) pulses/8 h (P less than 0.05), after clobazam. Mean LH concentrations and peak amplitudes did not change significantly. These results suggest that clobazam mediates its effects on LH secretion at the hypothalamic level by increasing the frequency of episodic GnRH release. PMID- 2566249 TI - Involvement of histaminergic neurons in the stress-induced release of pro opiomelanocortin-derived peptides in rats. AB - Histamine, which acts as a neurotransmitter, stimulates the release of the pro opiomelanocortin derived peptides ACTH, beta-lipotropin, and beta-endorphin. Since stress affects the hypothalamic turn-over of neuronal histamine, we investigated the role of histaminergic neurons in the mediation of the stress induced release of ACTH and beta-endorphin immunoreactivity in male rats. In control animals histamine receptor antagonists had no effect on the release of ACTH or beta-endorphin immunoreactivity. Restraint and ether stress increased plasma ACTH 3- and 2-fold, respectively. The responses were almost prevented by intracerebroventricular or intra-arterial infusion of the H2-receptor antagonists cimetidine and ranitidine. Infused intracerebroventricularly the H1-receptor antagonist mepyramine inhibited the ACTH response to restraint by 45% (P less than 0.01), but had no effect on the response to ether. Infused intra-arterially the H1-receptor antagonists mepyramine or SKF-93944 had no effect. Restraint and ether stress increased plasma beta-endorphin immunoreactivity 6- and 5-fold, respectively. Sephadex G-50 gel chromatography of plasma showed that the beta endorphin immunoreactivity in stressed rats co-eluted with beta-lipotropin and beta-endorphin, whereas the immunoreactivity in control animals co-eluted almost exclusively with beta-endorphin. The H2-receptor antagonists cimetidine and ranitidine infused intracerebroventricularly inhibited the responses of beta endorphin immunoreactivity to restraint and ether stress by 90 and 70%, respectively, whereas intra-arterial infusion of these antagonists inhibited the responses by only 50 and 60%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566250 TI - Midazolam does not potentiate the effect of vecuronium in patients. AB - Reports of midazolam interaction with vecuronium in animals prompted us to compare midazolam (0.25 mg kg-1) with thiopentone (5 mg kg-1) for possible interactions with vecuronium in patients, when used for induction of anaesthesia. After the administration of either of the two induction agents, the patients received vecuronium 0.1 mg kg-1. The onset time, duration of action and 25-75% recovery index of the neuromuscular blockade were recorded by measuring the force of thumb adduction evoked by ulnar nerve stimulation. We found no differences between patients receiving either midazolam or thiopentone in their response to vecuronium. In three of the ten patients receiving midazolam, the injection of this drug produced a 8-29% reduction of the initial twitch height. PMID- 2566251 TI - Spontaneous recovery of residual neuromuscular blockade after atracurium or vecuronium during isoflurane anaesthesia. AB - With atracurium and vecuronium, spontaneous recovery of residual neuromuscular blockade monitored electromyographically during 0.5% isoflurane anaesthesia was studied in 60 patients undergoing plastic surgery. After thiopentone, in random order, either atracurium 0.5 mg kg-1 or vecuronium 0.1 mg kg-1 was administered and isoflurane added to N2O and O2 mixture. Following spontaneous recovery of both the single twitch amplitude (T1) to 75% of the control value and the train of-four ratio (TOF ratio) to 75%, incremental doses of the relaxant were given to maintain the T1 at less than 10%. Before the end of surgery, the blockade was again permitted to recover spontaneously. During the initial spontaneous recovery, the mean recovery time of T1 from 25% to 75% (the recovery index) with atracurium was longer (P less than 0.001) than that with vecuronium (13.2 min and 10.1 min, respectively) but, during the second recovery, the mean recovery index was shorter (P less than 0.05) with atracurium than with vecuronium (16.1 min and 19.8 min, respectively). The recovery time from T1 75% to TOF ratio 75%, indicating the recovery rate of residual neuromuscular blockade, with atracurium was about 15 min after both the initial and the second recoveries. With vecuronium, the respective recovery times were significantly (P less than 0.001) longer (25.6 min and 38.5 min, respectively). It is concluded that with vecuronium there is slower spontaneous recovery of residual neuromuscular blockade than with atracurium. PMID- 2566253 TI - The influence of 0.5% isoflurane on a vecuronium-induced neuromuscular blockade. AB - The influence of adding 0.5% isoflurane to a narcotic-based anaesthesia on the duration of effect and recovery time after repetitive administration of vecuronium was studied in ten healthy patients. The twitch response in the adductor pollicis muscle was recorded after supramaximal train-of-four (TOF) stimulation of the ulnar nerve at the wrist. Prior to endotracheal intubation a bolus dose of vecuronium (0.08 mg/kg b.w.) was given. During surgery repeated injections of vecuronium (0.02 mg/kg b.w.) were administered at a TOF ratio of 0.25. Hand-skin temperature, systolic blood pressure, end-tidal CO2 and isoflurane concentrations were continuously monitored. Before and after 90 min administration of isoflurane, the duration of effect was 21 +/- 4 and 24 +/- 5 min (mean +/- s.d., P less than 0.05) respectively. Corresponding recovery times were 270 +/- 60 and 280 +/- 70 s (n.s.). Skin temperature remained unchanged and systolic blood pressure showed only minor variations. The addition of 0.5% isoflurane to a narcotic-based anaesthesia causes a moderate increase in duration of effect but no change in recovery time from a repetitive vecuronium-induced neuromuscular blockade of 0.02 mg/kg. PMID- 2566252 TI - Cardiovascular effects of vecuronium, atracurium, pancuronium, metocurine and RGH 4201 in dogs. AB - The effect on the cardiovascular haemodynamic status of five neuromuscular blocking drugs, RGH-4201, vecuronium, atracurium, pancuronium and metocurine, was studied in five conditioned foxhounds anaesthetised with fentanyl. Changes in heart rate, mean arterial blood pressure, central venous pressure, mean pulmonary artery pressure, pulmonary capillary wedge pressure, and cardiac output were recorded at 2, 5, 10, 20 and 30 min after administration of the drugs. From these, stroke volume, systemic vascular resistance and pulmonary vascular resistance were calculated. Administration of RGH-4201 was followed by a pronounced increase in heart rate, accompanied by an increase in cardiac output and a decrease in systemic and pulmonary vascular resistance. Metocurine and pancuronium resulted in a decrease of right and left filling pressures and systemic-/pulmonary vascular resistance. Changes after atracurium, vecuronium and metocurine were minimal. It is concluded that RGH-4201 causes major alterations in the cardiovascular haemodynamic status in dogs anaesthetised with fentanyl when compared to vecuronium, atracurium, metocurine and pancuronium. With respect to cardiovascular stability, atracurium and vecuronium offer advantages. PMID- 2566254 TI - Myoclonic encephalopathy due to bismuth salts: treatment with dimercaprol and analysis of CSF transmitters. AB - Two cases of myoclonic encephalopathy due to bismuth salts intoxication are reported. In both, treatment with dimercaprol led to clinical recovery. This therapy was shown to enhance bismuth clearance. We also present data on the CSF metabolites dopamine, norepinephrine and serotonin of one patient. PMID- 2566255 TI - Alzheimer's disease: areal and laminar pathology in the occipital isocortex. AB - Sensitive and specific silver methods for demonstration of (1) amyloid and/or precursors of amyloid and (2) neurofibrillary changes were applied to examine the pathology revealed by the occipital isocortex in cases of Alzheimer's disease and age-matched controls. In general, amyloid and/or precursors of amyloid are encountered in plaque-like formations. Large numbers of amyloid plaques occur in layers that only occasionally harbor neuritic plaques. Amyloid deposits can be found in abundance in the occipital cortex of demented individuals exhibiting an only sparse number of neuritic plaques. In demented individuals the striate area contains almost as much amyloid as the parastriate area or the peristriate region. Neurofibrillary changes are encountered in neuritic plaques, neurofibrillary tangles, and neuropil threads. Neuritic plaques are predominantly found in layers II and III. Their density changes even within the boundaries of architectonic units. Large numbers of plaques are found in the cortex covering the depth of the sulci. The number of neurofibrillary tangles increases abruptly when passing the striate/parastriate and the parastriate/peristriate boundaries. The neuropil threads may densely fill a layer without the presence of neurofibrillary tangles (layer V of the striate area). Neuropil threads contribute a substantial part to the total amount of the intraneuronally deposited pathological material. PMID- 2566256 TI - [Loprazolam: a new imidazobenzodiazepine for the treatment of insomnia]. AB - Loprazolam is a 1,4-benzodiazepine with hypnotics properties, useful for acute or chronic insomnia. His half-life of 7 to 8 hours in healthy adults, it may have advantages over benzodiazepines of short and longer-acting hypnotics. Although Loprazolam has been shown to have anti-anxiety, anticonvulsant and muscle relaxant effects, a longer acting benzodiazepine may be more appropriate when this prolonged effects is desired. Loprazolam would appear how a effective hypnotic with minimal side effects, which come to cover a place between the benzodiazepines with short and long half-life. PMID- 2566257 TI - Genetics of two populations of Glossina morsitans centralis (Diptera: Glossinidae) from Zambia. AB - Glossina morsitans centralis Machado was collected from the main fly belt west of Mumbwa Zambia and from the apparently isolated 'Keembe pocket' and 11 gene-enzyme systems were examined by polyacrylamide gel electrophoresis. There were no significant differences in allele frequencies among flies collected entering a vehicle, from fly-rounds, or from F3 traps in the main fly belt. Mean heterozygosity per locus is slightly higher in flies from the main fly belt than it is in flies from the 'Keembe pocket'. Allele frequencies at loci for xanthine oxidase (Xo), aldehyde oxidase (Ao) and a thoracic esterase (Est-2) were significantly different in the two populations and it is concluded that there is little gene flow between them. PMID- 2566258 TI - Contribution of focal haemorrhage and microvascular fibrin deposition to fatal envenoming by Russell's viper (Vipera russelli siamensis) in Burma. AB - In Burma, clinicopathological studies were carried out in three young farmers who died 15, 52 and 36 h after being bitten by Russell's vipers. Clinical features included local swelling, spontaneous systemic bleeding, defibrination, shock, cardiac arrhythmia, hypoglycaemia, coma and oliguria. On admission to hospital, 15, 48 and 21 h after the bites, serum venom antigen concentrations ranged from 50 to 130 ng/ml. Autopsies revealed widespread congestion and bleeding in the lungs, gastrointestinal and renal tracts, adrenals, heart, brain and anterior pituitary. There was histopathological evidence of focal haemorrhage and fibrin deposition at the site of the bite and in the pituitary, lungs and kidneys and acute tubular necrosis. Deposition of fibrin microthrombi results from the action of venom procoagulants. Shock was attributed to increased capillary permeability, revealed clinically by conjunctival oedema. Acute pituitary/adrenal failure in one case was explained by fibrin deposition and haemorrhage in the anterior pituitary--resembling Sheehan's syndrome. Acute tubular necrosis resulted from ischaemia caused by fibrin deposition and to prerenal factors. An intractable cardiac tachyarrhythmia may have been caused by subendocardial and myocardial haemorrhages. PMID- 2566259 TI - Cytochemical demonstration of lectin binding-sites in the cuticle and tissues of Acanthocheilonema viteae (Filarioidea). AB - The lectin-gold technique was used for the ultrastructural localization of lectin binding sites on thin sections of Lowicryl K4M embedded adult females, infective larvae and SDS-2-mercaptoethanol-insoluble cuticle components of Acanthocheilonema (Dipetalonema) viteae. Helix pomatia lectin (HPL) coupled to 14 nm gold particles, was used for the demonstration of N-acetyl-D-galactosamine containing glycoconjugates. Triticum vulgaris (wheat germ) agglutinin (WGA) coupled to 10 nm gold particles after cross-linking to BSA or ovomucoid-gold after application of unlabeled WGA, demonstrated WGA binding sites (N-acetyl-D glucosamine). With both lectins no surface labelling of the cuticle was observed, but subcuticular layers reacted positively. HPL-gold was bound to cuticular fibers, the matrix and to the electron dense layer within the cortical zone of the cuticle of female worms. WGA-gold complexes were bound mainly to the cuticle matrix and somatic tissues. The results support the hypothesis that tissue dwelling parasitic nematodes have reduced their surface carbohydrates perhaps as a consequence of their parasitic life. PMID- 2566260 TI - Schistosoma rodhaini: intramolluscan larval development, migration and replication processes of daughter sporocysts. AB - The larval development of Schistosoma rodhaini in Biomphalaria glabrata presents two features of interest. The first concerns the migration of daughter sporocysts: only a fraction of the daughter sporocysts produced by the mother sporocyst migrate directly towards the digestive gland of the snail and become cercariogenous; the others remain in the anterior region of the snail where the majority transforms into small sporocystogenous sporocysts. The second point concerns the replication processes: only direct replication of daughter sporocysts was observed in sporocysts in a prehepatic localization. The demographic processes involved in the development of S. rodhaini are compared with other species of Schistosoma and interpreted as an adaptive character for better exploitation of the snail host. PMID- 2566261 TI - Distribution of surgical hydatidosis in central Tunisia (1982-1985). AB - The mean annual surgical incidence rates (MASIR) of hydatidosis were measured for the 1982-1985 period in central Tunisia in order to investigate the natural history of human hydatidosis and provide baseline data for the evaluation of future prevention campaigns. 986 cases were identified from surgical records of regional hospitals to which cases from central Tunisia are mandatorily referred. The overall MASIR was 19.3 per 10(5) inhabitants. The MASIR according to district varied from 0 to 56.6 per 10(5), was higher in women than in men (22.6 versus 15.8 per 10(5] and increased with age (maximum 53.0 per 10(5) in the 50-59 age group for women and 27.1 per 10(5) in the 40-49 group for men). Among the 940 cases with single organ hydatidosis, liver ranged first (55.3%, 545/940), then lung (32.4% 320/940), kidney (2.7%) and spleen (1.8%). Among the 865 subjects with single lung or liver hydatidosis, liver was more often involved in women (349 out of 503) than in men (196 out of 362) (chi 2 = 20.9, p less than 0.001). In both sexes, the lung/liver ratio decreased with age. These data reveal the existence of highly endemic foci of hydatidosis within central Tunisia. The predominance of infection in women might be due to sex related behavioral differences. The causes of liver predominance in women and variation of lung/liver ratio are open to question. PMID- 2566262 TI - Use of DNA probes to identify Trypanosoma congolense and T. simiae in tsetse flies from The Gambia. AB - Species- and strain-specific DNA probes were used to identify patent midgut infections in Glossina morsitans submorsitans and G. palpalis gambiensis captured at four sites in The Gambia. 52% of mature Nannomonas infections and 12% of immature infections were identified. Trypanosoma (Nannomonas) simiae accounted for the majority of identified infections in G.m. submorsitans, indicating the importance of distinguishing this species from the closely related T.(N) congolense when assessing the trypanosomiasis challenge to cattle. Both the savannah and riverine-forest groups of T. congolense were present, although the riverine-forest form was found only in G.p. gambiensis at Pirang, an isolated area of forest. Two-thirds of the samples remain unidentified by probes specific for: Trypanozoon; T. congolense savannah, riverine-forest and Kenya coast forms; T. simiae; and T. vivax, probably owing in part to low numbers of trypanosomes. However, the failure to identify several heavy Nannomonas infections, strongly suggests the presence of a further, as yet unknown, kind of Nannomonas. PMID- 2566263 TI - Chitin derivatives as novel substrates for Trypanosoma brucei brucei attachment in vitro. AB - Procyclic trypomastigotes of Trypanosoma brucei brucei, cultured in Cunningham's medium with 20% heat-inactivated foetal calf serum at 27 degrees C, attached to chitosan and to gels of N-acetyl chitosan and glycol chitosan. Following attachment, epimastigotes, metacyclic-like trypomastigotes and multinucleate parasites appeared in the culture supernatant. PMID- 2566264 TI - Current considerations on a Loa loa simian reservoir in the Congo. PMID- 2566265 TI - A new method for the isolation of the trypanocidal factor from normal human serum. PMID- 2566266 TI - The spread of freshwater snails including those of medical and veterinary importance. AB - The present paper discusses the spread of freshwater snails that act as intermediate hosts for various trematodes that cause diseases in man and in cattle. Examples of snail species which have spread across natural barriers are reviewed as well as the mechanisms involved in this transport. Focus is put on the extensive trade in freshwater aquatic plants and aquarium fish, and the need for better control of this trade is emphasized in order to minimize the risk of unintended import of potentially harmful snails. PMID- 2566267 TI - Biological control of the snail hosts of Schistosoma mansoni in the Caribbean area using Thiara spp. AB - Field observations and experiments using thiarid snails as competitors of Biomphalaria spp., potential intermediate hosts of Schistosoma mansoni in the Caribbean area, are reviewed. The parthenogenetic snails, Thiara granifera and T. (= Melanoides) tuberculata, were introduced to the Neotropical area in recent decades. In numerous islands and countries, these oriental species have demonstrated their capacity to colonize rapidly and densely many types of habitats while at the same time reducing and even eliminating populations of Biomphalaria spp. The results of field experiments, carried out in several Caribbean islands, have shown the efficiency as well as the limitations of T. tuberculata as a competitor of B. glabrata and B. straminea. In St. Lucia, B. glabrata was apparently eliminated from marshes and streams, 6 to 22 months after the introduction of the competitor. In Martinique, T. tuberculata was introduced into two groups of water-cress beds which constituted the last transmission sites of schistosomiasis on the island. In just less than three years after the introduction of the competitor, both B. glabrata and B. straminea have been eliminated from the transmission sites. In Guadeloupe, several introductions have been carried out in different types of habitat such as permanent ponds, canals, streams and temporary marshes. The findings of all field experiments have indicated that thiarid snails as competitors of pulmonates are favoured by the presence of permanent and stable habitats, preferably shallow, with emergent plants and well oxygenated. On the other hand, the competitor snails are at a disadvantage in waterbodies which are temporary, extremely deep, poorly oxygenated or with a dense mat of floating aquatic vegetation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566269 TI - New observations on cyclical development of Trypanosoma vivax in Glossina. AB - It is widely held that cyclical development of Trypanosoma vivax in Glossina is confined to the proboscis. This view has been re-examined in a series of experiments. Teneral G. morsitans centralis were fed on a goat infected with T. vivax IL 1392 and dissected 1-2 h after feeding. The infection rates in the labrum and hypopharynx were 40% and 0%, in contrast to 82% and 58%, respectively, observed in a control group dissected on day 25. This suggested that in a significant number of tsetse, cyclical development of T. vivax was initiated at sites other than the proboscis. Subsequent experiments revealed the presence of trypomastigotes, pre-epimastigotes and epimastigotes in the cibarium/oesophageal region of tsetse dissected 1-48 h after an infected feed. To investigate this further, tsetse proboscides were excised at intervals beginning 1 h after an infected feed, and transferred to in vitro culture conditions. Parasite multiplication and full cyclical development were only observed in proboscides excised 4 h or later after the infected bloodmeal. Thus, it would appear that at least in a number of tsetse, T. vivax cyclical development initially occurs in the cibarium/oesophageal region from where parasites migrate to the food canal of the proboscis and development is completed to infective metatrypanosomes in the hypopharynx. PMID- 2566268 TI - Chloroquine-resistant Plasmodium falciparum in eastern Sudan. AB - In vivo testing of the sensitivity of Plasmodium falciparum to chloroquine was carried out in 61 falciparum malaria patients with acute symptoms, in Eastern Sudan. In 26 patients (42%), P. falciparum was resistant to chloroquine. Nine patients (15%) had RI resistance, seven (11%) had RII resistance while ten (16%) had RIII resistance. The persistance of parasitaemia and symptoms were highly correlated in patients with RIII responses. In 21 patients in vitro testing of chloroquine sensitivity was carried out simultaneously with the in vivo testing using the World Health Organization microtest. In vivo and in vitro testing were also highly correlated. Isolates from 12 patients with proven in vivo resistance, grew in vitro in the presence of chloroquine concentrations above 0.8 X 10(-6) mol/l blood. Resistant strains have either been spread by refugees across the borders from Ethiopia or have developed indigenously. Mounting drug pressure, mass movement of non-immune refugees and loss of immunity among local inhabitants, due to the drought, are in favour of development of an indigenous focus. Epidemics with intense transmission caused by heavy rains following the drought could have greatly enhanced the emergence and spread of resistant strains. PMID- 2566270 TI - Survival estimates for adult Culex pipiens in the Nile Delta. AB - In an endemic area for Wuchereria bancrofti filariasis in the Nile Delta, survival of adult female Culex pipiens was estimated by parity rate, mosquito infection and infectivity rates. Infection rates and 4th instar larval populations, as well as infection and parity rates, were linearly correlated. Infectivity correlated only with parity rates. These associations corresponded to parallel changes in ambient temperature. Although survival calculated from parity rates measured longevity of both infected and non-infected Cx. pipiens, survival based on infection and infectivity was a more reliable indicator for parasite transmission. PMID- 2566271 TI - Seasonal population changes and malaria transmission potential of Anopheles pharoensis and the minor anophelines in Mwea Irrigation Scheme, Kenya. AB - A study in 1984 and 1985 showed that Anopheles gambiae s.l. and An. pharoensis were the major anophelines in Mwea Irrigation Scheme, Kenya, constituting 83.86% and 15.69% of the catch respectively. Four minor species made up the remaining 0.45%. The irrigation phase of the rice cultivation cycle in August, which linked the flooding effects of the two rainy seasons, resulted in major population increases of An. pharoensis and enabled continuous breeding for up to 9 months per year. The average of mean monthly proportions of unfed, bloodfed, and gravid females was 26.6, 58.8, and 14.6% respectively. The Plasmodium falciparum sporozoite rates for An. pharoensis were 1.3% by ELISA and 0.68% by dissection, while those for An. funestus were 1.7% by ELISA and 1.25% by dissection. An. pharoensis can contribute to the epidemiology of Malaria in the Mwea area. PMID- 2566272 TI - Isolation and cultivation in vitro to the infective, metacyclic stage of Trypanosoma (Nannomonas) simiae from Glossina morsitans submorsitans. AB - Two separate trypanosome isolations were made from a single Nannomonas-infected Glossina morsitans submorsitans from The Gambia. Inoculation of a piglet with the infected hypopharynx produced an infection with Trypanosoma simiae. DNA was isolated from the bloodstream forms to prepare a probe specific for this species. Trypanosomes isolated from the fly midgut were frozen in liquid nitrogen and then cultivated in vitro. Amplification of this population and elimination of a yeast contaminant were achieved by two passages through laboratory G. m. morsitans. Further cultivation in vitro resulted in the production of epimastigotes and, later, metacyclic forms. Two pigs inoculated with cultivated metacyclic forms developed infections with atypical, relapsing parasitaemias and extended survival time. Neither the metacyclic forms, nor bloodstream forms derived from them, infected calves. The identity of various stages of the in vitro cultivated, procyclic-derived stock was confirmed morphologically and with the T. simiae specific DNA probe. PMID- 2566273 TI - A simple and rapid method to initiate Trypanosoma brucei brucei and T. brucei evansi bloodstream form cultures. AB - Initiation of cultures of bloodstream forms of trypanosomes from the Trypanozoon subgenus is an established laboratory procedure. The trypanosomes are usually separated from the blood cells of the donor animal by centrifugation on a density gradient (Hirumi et al., 1977) or by differential centrifugation (Hill et al., 1978; Brun et al., 1981; Zweygarth et al., 1982; Baltz et al., 1985). Isolation on an anion exchange column (Lanham and Godfrey, 1970) has been little used for this purpose. We describe a simple and rapid method for the initiation of trypanosome bloodstream form cultures from infected host blood, avoiding centrifugation and anion exchange column procedures. Two T.b. brucei stocks, CP 271 and CP 547, and two T.b. evansi stocks, CP 1134 and CP 2087, were used. Stock CP 271 was isolated in 1980 from a goat in Matuga/Kenya, stock CP 547 was isolated in 1985 from a naturally infected bovine in Jilib/Somalia. The two T.b. evansi stocks were both isolated from camels, stock CP 1134 in 1979 in Kulal/Kenya, stock CP 2087 in 1980 in the Sudan. PMID- 2566275 TI - Acute transient memory loss. AB - Anterograde amnesia refers to the inability to form new memories. Alcoholic blackout, benzodiazepine-induced amnesia and transient global amnesia are three disorders that result in acute transient memory loss. These disorders may be difficult to recognize, because the memory loss is not usually accompanied by other symptoms of neurologic impairment. PMID- 2566274 TI - Sulphasalazine-induced reversible male infertility. AB - A male patient suffering from ulcerative colitis, presented with primary infertility due to sulphasalazine therapy. Sulphasalazine was discontinued and the patient was treated with a 5-aminosalicylic acid preparation (Salofalk) in the form of an enema. After 3 months, the semen quality was dramatically improved and a successful pregnancy ensued. The sulphapyridine moiety of sulphasalazine seems to be responsible for male infertility and depressed semen quality. The metabolite 5-aminosalicylic acid is proposed as a suitable alternative to sulphasalazine in cases of male infertility. PMID- 2566276 TI - The current state of pertussis and pertussis vaccines. A report on the Fifth International Symposium on Pertussis. PMID- 2566277 TI - Cooperative regional program for distribution of poison prevention information and syrup of ipecac. AB - A regional program to distribute poison prevention information and syrup of ipecac to families that have regular contact with young children is described. In December 1985 the department of pharmacy services at Bristol Hospital in Connecticut proposed implementation of a poison prevention program targeted to families with young children (less than 12 years of age) in the hospital's five town service area. A planning committee was created to define program goals and oversee operations. The committee decided that poison prevention kits consisting of an instructional booklet and a one-ounce bottle of syrup of ipecac would be distributed to selected residents of the five-town area, with individual instruction provided in the correct use of syrup of ipecac. Funding was provided principally by the hospital, with some additional money from private foundations. The project was named CAP (Combating Accidental Poisoning) and ran initially from December 1, 1986, to June 30, 1987. Kits were distributed in cooperation with area health-care professionals with whom families had regular contact, including pediatricians, family-practice physicians, and community pharmacies. Apart from the hospital pharmacy service itself, the most effective participants were pediatricians; family-practice physicians were highly ineffective. During the course of the initial CAP program 6610 kits were distributed, with 48.7% going to families considered at high risk for the occurrence of an accidental child poisoning. An ongoing program to distribute these kits to all newly delivered mothers and to all area pediatricians free of charge has resulted in more than 15,000 kits being distributed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566278 TI - Characteristics of ANP-sensitive guanylate cyclase in inner medullary collecting duct cells. AB - Our previous characterization of equilibrium binding kinetics of atrial natriuretic peptide (ANP) to the surface of inner medullary collecting duct (IMCD) cells suggested the existence of a single class of high-affinity receptors, functionally coupled to increases in cellular guanosine 3',5'-cyclic monophosphate (cGMP). We have now sought to understand the mode of regulation of this signal transduction system by studying the particulate guanylate cyclase (PGC) enzyme from these cells. PGC activity with and without ANP in membranes, made by homogenization and high-speed centrifugation of suspensions of IMCD cells, was linear up to 5 min and was stimulated by ANP [143 +/- 21 (ANP) vs. 38 +/- 7 (control) pmol/mg protein, n = 3, P less than 0.02]. Vmax increased more than threefold with ANP [130 +/- 19 (ANP) vs. 35 +/- 4 (control) pmol.mg protein 1.min-1, n = 4, P less than 0.005] without significant change in the Km [0.68 +/- 0.17 (ANP) vs. 0.55 +/- 0.08 (control) mM] of the enzyme. Half-maximal stimulation of guanylate cyclase activity occurred at 5 x 10(-10) M ANP, a concentration consistent with our binding data, and with physiological effect. PGC required divalent cations for basal activity and for ANP-stimulated activity; Mg2+ and Mn2+ were most potent in this respect, and Ca2+ was without effect. Both basal and stimulated PGC activities were inhibited in response to changes in the NaCl, but not urea concentration of the assay system. We conclude that binding to the single 120-130 kDa ANP receptor in IMCD cells results in stimulation of PGC by increasing its Vmax and thereby elevating intracellular cGMP, the likely mediator of ANP action in these cells. PMID- 2566279 TI - Cystine and lysine reabsorption in the isolated perfused rat kidney. AB - Renal tubular reabsorption of cystine and lysine were studied in the isolated perfused rat kidney to bridge the gap between in vivo clearance studies, and in vitro transport studies of tubule fragments, cells, and brush-border membranes. Lysine was reabsorped by a saturable transport system shared by the dibasics. Cystine was also reabsorbed by a saturable transport system, which was shared in part by the dibasics (maximum inhibition 30%). The lysine threshold (Fmin) was 0.9 mumol.min-1.g-1, with a tubular maximum (TM) of 2.4 mumol.min-1.g-1. The cystine Fmin was 0.06 mumol.min-1.g-1; the TM could not be estimated because it was above the limit of cystine solubility. There was no evidence of cystine "secretion." The gamma-glutamyltransferase inhibitor, AT-125, decreased cystine excretion, but only in the presence of glutathione, glycine, glutamate, and the diabasic amino acids. This suggests that cystine from glutathione degradation at the brush border may contribute to urinary cystine (an explanation of the phenomenon of cystine secretion), but only under certain conditions. PMID- 2566280 TI - Pentobarbital anesthesia alters pulmonary vascular response to neural antagonists. AB - We investigated the effects of pentobarbital sodium anesthesia on vasoregulation of the pulmonary circulation. Our specific objectives were to 1) assess the net effect of pentobarbital on the base-line pulmonary vascular pressure-to-cardiac index (P/Q) relationship compared with that measured in conscious dogs, and 2) determine whether autonomic nervous system (ANS) regulation of the intact P/Q relationship is altered during pentobarbital. P/Q plots were constructed by graded constriction of the thoracic inferior vena cava, which produced stepwise decreases in Q. Pentobarbital (30 mg/kg iv) had no net effect on the base-line P/Q relationship. In contrast, changes in the conscious intact P/Q relationship in response to ANS antagonists were markedly altered during pentobarbital. Sympathetic alpha-adrenergic receptor block with prazosin caused active pulmonary vasodilation (P less than 0.01) in conscious dogs but caused vasoconstriction (P less than 0.01) during pentobarbital. Sympathetic beta-adrenergic receptor block with propranolol caused active pulmonary vasoconstriction (P less than 0.01) in both groups, but the magnitude of the vasoconstriction was attenuated (P less than 0.05) during pentobarbital at most levels of Q. Finally, cholinergic receptor block with atropine resulted in active pulmonary vasodilation (P less than 0.01) in conscious dogs, whereas vasoconstriction (P less than 0.01) was observed during pentobarbital. Thus, although pentobarbital had no net effect on the base-line P/Q relationship measured in conscious dogs, ANS regulation of the intact pulmonary vascular P/Q relationship was altered during pentobarbital anesthesia. PMID- 2566281 TI - A novel effect of norepinephrine on cardiac cells is mediated by alpha 1 adrenoceptors. AB - In the heart, alpha-adrenergic agonists have long been known to produce a positive inotropic effect that is rate dependent and associated with action potential prolongation but is not accompanied by adenosine 3',5'-cyclic monophosphate (cAMP) elevation. The ionic mechanism of these effects is unknown. We report that a transient outward K+ current, a major determinant of plateau duration in rabbit and human atria, is strongly inhibited by norepinephrine and the alpha-adrenoceptor agonists methoxamine and phenylephrine. These effects of alpha-stimulation can be blocked by prazosin. The reduction in the transient outward current substantially slows action potential repolarization. These results can explain the regional and species-dependent positive inotropic effects of alpha-adrenergic stimulation in the heart and give important new insight into the autonomic regulation of cardiac function. In addition, reduction in this repolarizing current during the enhanced alpha-adrenergic responsiveness of myocardial ischemia may be a factor in the genesis of arrhythmias produced by catecholamines. PMID- 2566282 TI - Plasma atriopeptin during exercise in dogs under beta-blockade. AB - The purpose of this study was to describe plasma atriopeptin concentrations at rest and in response to moderate treadmill exercise (10 min, 4 km/h, 26% slope) performed with or without nonspecific beta-adrenergic blockade (1 mg/kg iv propranolol) in 10 mongrel dogs [19 +/- 2 (SE) kg]. A small (20%) but significant (P less than 0.05) increase in plasma atriopeptin concentration was observed from rest (43 +/- 5 pg/ml) to exercise (52 +/- 6 pg/ml) without beta-blockade. Propranolol significantly reduced heart rate at rest (89 +/- 7 vs. 104 +/- 7 beats/min) and during exercise (96 +/- 10 vs. 176 +/- 11 beats/min), and this was associated with a larger increase in plasma atriopeptin concentration during exercise (rest 46 +/- 6 pg/ml; exercise 171 +/- 22 pg/ml). Exercise under beta blockade is associated with an increased preload of the heart. These results further support the hypothesis that atriopeptin release during exercise is under the control of atrial stretch. The higher plasma atriopeptin concentration observed during exercise under beta-blockade may contribute to the reduction of the response of plasma renin activity (1.0 +/- 0.1 vs. 3.0 +/- 0.6 ng.ml-1.h-1) and aldosterone concentration (87 +/- 36 vs. 138 +/- 25 pg/ml). Vasopressin concentration was lower at rest and during exercise under propranolol (3.5 +/- 1.3 vs. 4.9 +/- 0.9 and 6.1 +/- 2.2 vs. 9.9 +/- 1.5 pg/ml, respectively), which might reflect a dissociation between activity of the renin-angiotensin system and vasopressin release. PMID- 2566283 TI - Autonomic modulation of ultradian blood pressure and heart rate oscillations in dogs. AB - Autonomic receptor modulation of ultradian oscillations of blood pressure and heart rate was studied in telemetered free-running dogs. Data, analyzed for their harmonic content by fast Fourier transform (FFT) methods, indicated that ultradian and circadian oscillations of 22.9 +/- 2.5 and 10.5 +/- 0.9 (SD) mmHg, respectively, were present. The average principal frequency for the ultradian oscillations in 12 dogs was 0.760 +/- 0.11 cycles/h for arterial pressure and 0.808 +/- 0.10 for heart rate. Atropine had no effect on periodicity of either arterial pressure or heart rate. Metoprolol, a beta 1-antagonist, or hexamethonium, a ganglionic blocker, significantly reduced the power of both arterial pressure and heart rate (P less than 0.05), whereas the primary frequencies of both were unchanged. Prazosin, an alpha 1-blocker, sharply reduced arterial pressure power (P less than 0.05) and increased the power of heart rate (P less than 0.05), demonstrating that it is possible to uncouple arterial pressure oscillations from influences of heart rate. We conclude that the sympathetic limb of the autonomic nervous system is primarily responsible for these oscillations and that vagal influences on the heart partially dampen these rhythms. PMID- 2566284 TI - Bradycardia in patients receiving atracurium or vecuronium in conditions of low vagal stimulation. AB - Four groups of 20 patients each received either vecuronium or atracurium together with either glycopyrronium or saline, and underwent anaesthesia free of vagolytic drugs, and surgery devoid of vagal activity. Determinations of plasma histamine concentrations were made to examine the possible correlation between these levels and changes in heart rate and blood pressure as well as a possible relationship with skin reactions after the administration of the relaxants. Patients who received vecuronium without the anticholinergic drug, glycopyrronium, showed a greater tendency towards bradycardia (though not statistically significant) than those given atracurium. More cutaneous reactions were observed with patients who received atracurium than in those with vecuronium, but there was no correlation with plasma histamine concentrations of either relaxant group. There was no correlation either between histamine concentrations and heart rate or blood pressure associated with atracurium. The incidence of bradycardia with either relaxant is low if the anaesthetic technique and the surgery are devoid of vagal activity. PMID- 2566285 TI - Respiratory arrest after a caudal injection of bupivacaine. AB - A 3.5-year-old child with Cornelia de Lange syndrome presenting for orchidopexy and herniotomy received general anaesthesia which was supplemented by a caudal injection of bupivacaine. Shortly after this he had a respiratory arrest with fixed dilated pupils. No cardiovascular system instability was observed. Approximately one hour later the child made a rapid and complete recovery. The possible causes of these events are discussed with particular reference to the potential effects of caudal injections on intracranial pressure. PMID- 2566286 TI - [The clinical significance of drug interactions between opiates and calcium antagonists. A randomized double-blind study using fentanyl and nimodipine within the framework of postoperative intravenous on-demand analgesia]. AB - It is widely accepted that the nociceptive state and opiate-induced nociception are regulated at least in part by calcium ions. Animal experiments suggest that systemically or intracerebroventricularly applied calcium antagonizes analgesic effects, whereas calcium chelating agents or calcium channel blockers enhance them. Recently, von Bormann et al. [3] reported a fentanyl-saving effect in cardiovascular patients who had received an intraoperative infusion of nimodipine; this finding was discussed as a possible synergistic analgesic interaction. Since doubts remained as to whether this interpretation was justified, the present study aimed to verify, in awake postoperative patients, whether nimodipine increased the analgesic efficacy of fentanyl. Forty ASA I-II patients (mean age 43-44 years) undergoing elective hysterectomy under standardized balanced anesthesia were investigated. In the recovery room, they were allowed to self-administer fentanyl by means of the On-Demand Analgesia Computer (ODAC). Demand dose was 34.5 micrograms, infusion rate 4 micrograms/h, lockout time 1 min, hourly maximum dose 250 micrograms. The patients were randomly and double-blindly assigned to have an additional infusion of either placebo (P) or nimodipine (N: 15 micrograms/kg/h during the first 2 h, 30 micrograms/kg/h from the 3rd to the 12th h). Fentanyl consumption, pain scores (actual and retrospective), blood pressure, heart rate, respiratory rate, and side-effects were monitored. The mean duration of patient-controlled analgesia was 16 (P) to 19 (N) h, during which time 0.64 +/- 0.46 (N) to 0.79 +/- 0.43 (P) micrograms fentanyl/kg/h was demanded. Pain relief was very satisfactory in 92.5% of the patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566287 TI - Double-blind comparison of midazolam and temazepam as oral premedicants for outpatient anaesthesia. AB - Oral premedication with midazolam 7.5 mg was compared with temazepam 20 mg in a double-blind study of sixty patients undergoing day-stay urological surgery. One hour following ingestion similar degrees of anxiolysis and sedation were reported by patients for both compounds. However, midazolam was observed by anaesthetists to produce the greater anxiolytic effect and was given the better overall assessment. Midazolam produced significantly greater amnesia both at the time of induction and 30 minutes postoperatively. At the time of discharge four hours postoperatively no significant difference could be observed in psychomotor performance or subjective sedation although on the evening of surgery the temazepam group had a greater incidence of sleepiness and an earlier time to retiring. Although the differences were small, the residual post-discharge effects of temazepam lead us to conclude that midazolam 7.5 mg is the more suitable premedicant for outpatient use. PMID- 2566288 TI - Intrathecal somatostatin in cat and mouse studies on pain, motor behavior, and histopathology. AB - Effects of intrathecal (i.t.) somatostatin (SST) on nociception, motor function, and spinal cord pathology were evaluated in cats and mice. Cats chronically implanted with i.t. lumbar catheters received either a single injection of 2 mg SST i.t. (group I, N = 4), four repetitive injections on consecutive days of 2 mg SST i.t. (group II, N = 4) or saline i.t. (group III, N = 2). No analgesic effects were observed following single or repeated SST injections as evaluated by the skin twitch response. However, significant impairment of hind leg motor function ranging from unbalanced gait to paralysis was observed following the first SST i.t. injection. Histological examination of spinal cords six days after the first SST injection in group II showed multiple pyknotic neurons in all cats. Some cats showed focal demyelination in the posterior column of the spinal cord white matter. Mice received a single percutaneous injection of 50 micrograms SST i.t. (group I, N = 7), 5 micrograms SST i.t. (group II, N = 3), or saline i.t., (group III, N = 5). No analgesic effects were observed in groups II and III as evaluated by the hot plate (HP) and tail flick (TF) tests. Injection of 50 micrograms SST i.t. (group I) caused reversible flaccid hind leg paralysis in all mice and concomitant increases in HP and TF latencies. Histologic examination revealed focal demyelination in the spinal cord in three out of seven mice in this group. Present data substantiate neurotoxic effects of i.t. SST and lack of behaviorally defined antinociception at innocuous dosages. PMID- 2566289 TI - Resistance to muscle relaxants in a patient receiving prolonged testosterone therapy. PMID- 2566290 TI - Esmolol use during resection of pheochromocytoma: report of three cases. PMID- 2566291 TI - Assessment of compliance in children using inhaled beta adrenergic agonists. AB - The use of inhaled beta adrenergic agonists has assumed a greater role in the treatment of pediatric asthma as medications with a longer duration of action and more specificity for the beta 2 receptor have become available. Because of their effectiveness and safety, they are being selected as the drug of choice for maintenance therapy in children with asthma. The following study assessed compliance in asthmatic children using metaproterenol administered via a metered dose inhaler (MDI) on a daily basis. Compliance was determined in 17 asthmatic children (aged 5 to 13 years) by measurement of cannister weight before and after a 2-week treatment period. Five children were studied for one 2-week period, while 12 children were studied for two consecutive 2-week periods. Two of five children (40%) completing 2 weeks were compliant, while only one of 12 children (8%) completing 4 weeks of the study was determined to be compliant as judged by changes in cannister weight during the treatment period. We conclude that despite the efficacy and safety of beta 2 adrenergic agonists administered via a MDI, poor compliance could potentially detract from their value as a maintenance medication in some asthmatic children. PMID- 2566292 TI - The natural history and response to therapy of chronic urticaria and angioedema. AB - Previous reports have suggested that the etiology of chronic urticaria/angioedema (greater than 6 weeks) can be identified 10% to 30% of the time while few reports have addressed the natural history of chronic urticaria/angioedema. An analysis of all patients referred to the authors' practice between 1983-1985 with a diagnosis of urticaria/angioedema was performed. Patients with hereditary angioedema were excluded. Eighty-six of the 214 patients had chronic urticaria/angioedema. In the remaining 128 cases of acute urticaria there were four exercise induced, nine contact, six cold induced, six drug induced, 11 food induced, one viral hepatitis associated, 29 with dermographism, and 62 undetermined. An etiology could not be determined in any of the patients with chronic urticaria/angioedema. Laboratory tests, including CBC, chemistry panel, urinalysis, ANA, rheumatoid factor, complement studies, sedimentation rate, and skin tests were all noninformative. Chronic angioedema without urticaria occurred in only nine cases, 31 cases had chronic urticaria alone, and 46 cases had both chronic urticaria and angioedema. Of the patients followed over the 3-year period, 27 resolved while 48 continued to have urticaria/angioedema. Response to medications was variable and will be discussed. Our study suggests that an etiology is determined in much less than 10% of patients with chronic urticaria; fortunately, 32% of our cases resolved over a 3-year period. PMID- 2566293 TI - Long-term treatment with thymomodulin reduces airway hyperresponsiveness to methacholine. AB - We investigated the effect of thymomodulin, a calf thymus acid lysate with immunomodulating activity, on bronchial hyperresponsiveness to methacholine of atopic subjects with asthma. In 16 subjects we measured airway responsiveness at 30, 60, and 90 days after treatment with placebo (eight subjects) or thymomodulin (eight subjects; 80 mg daily orally). The degree of bronchial responsiveness to methacholine was significantly reduced at 90 days during treatment with thymomodulin and remained reduced, even if not significantly, 60 days after cessation of treatment. PMID- 2566294 TI - Functional interactions between TRH and 5-hydroxytryptamine (5-HT) and proctolin in rat brain and spinal cord. PMID- 2566295 TI - Use of receptor antagonist in elucidating the mechanism of action of TRH in GH3 cells. PMID- 2566296 TI - Inhibitors of TRH-degrading enzymes. PMID- 2566297 TI - Increased ciliary beating frequency of nasal mucosa following immunotherapy for allergy. AB - The present study was designed to elucidate the effect of immunotherapy on the beating frequency of nasal cilia in patients with nasal allergy. Of 40 patients with nasal allergy due to Dermatophagoides farinae, 20 were treated with immunotherapy by the use of D farinae extracts, and 20 control patients were treated with antihistamine tablets. Mucosal pieces were taken from the right inferior turbinate before and 1 year after the initiation of treatment, and the ciliary beating frequency (CBF) was examined by a photoelectric method. The use of antihistamine tablets did not increase CBF even when it relieved the nasal symptoms. Immunotherapy, on the other hand, increased CBF in 66.7% of patients when it alleviated their nasal symptoms. The CBF before immunotherapy of patients who showed an increase CBF after treatment was statistically higher than that of patients who did not. PMID- 2566298 TI - Carbohydrate metabolism in the mosquito pathogen Bacillus sphaericus 2362. AB - Bacillus sphaericus 2362 is pathogenic for mosquito larvae and is being considered for large-scale production as a larvicide. The inability of the bacteria to metabolize carbohydrates requires that they be grown on proteinaceous media. This bacterium was found to be unable to transport glucose or sucrose into the cell, and it lacked glucokinase and hexokinase activity. In addition, it lacked phosphoglucose isomerase, phosphofructokinase, and glucose 6-phosphate dehydrogenase, which are early enzymes of the Embden-Myerhof-Parnas and hexose monophosphate pathways. The presence of other enzymes in these pathways was indicated by assay, by the metabolism of glycerol to acetate, and by growth on acetate and gluconate as sole carbon sources. Critical enzymes of the Entner Doudoroff pathway were also shown to be absent. PMID- 2566300 TI - Cimetidine and beta-blockers. PMID- 2566299 TI - Enzymic analysis of the crabtree effect in glucose-limited chemostat cultures of Saccharomyces cerevisiae. AB - The physiology of Saccharomyces cerevisiae CBS 8066 was studied in glucose limited chemostat cultures. Below a dilution rate of 0.30 h-1 glucose was completely respired, and biomass and CO2 were the only products formed. Above this dilution rate acetate and pyruvate appeared in the culture fluid, accompanied by disproportional increases in the rates of oxygen consumption and carbon dioxide production. This enhanced respiratory activity was accompanied by a drop in cell yield from 0.50 to 0.47 g (dry weight) g of glucose-1. At a dilution rate of 0.38 h-1 the culture reached its maximal oxidation capacity of 12 mmol of O2 g (dry weight)-1 h-1. A further increase in the dilution rate resulted in aerobic alcoholic fermentation in addition to respiration, accompanied by an additional decrease in cell yield from 0.47 to 0.16 g (dry weight) g of glucose-1. Since the high respiratory activity of the yeast at intermediary dilution rates would allow for full respiratory metabolism of glucose up to dilution rates close to mumax, we conclude that the occurrence of alcoholic fermentation is not primarily due to a limited respiratory capacity. Rather, organic acids produced by the organism may have an uncoupling effect on its respiration. As a result the respiratory activity is enhanced and reaches its maximum at a dilution rate of 0.38 h-1. An attempt was made to interpret the dilution rate-dependent formation of ethanol and acetate in glucose-limited chemostat cultures of S. cerevisiae CBS 8066 as an effect of overflow metabolism at the pyruvate level. Therefore, the activities of pyruvate decarboxylase, NAD+- and NADP+-dependent acetaldehyde dehydrogenases, acetyl coenzyme A (acetyl-CoA) synthetase, and alcohol dehydrogenase were determined in extracts of cells grown at various dilution rates. From the enzyme profiles, substrate affinities, and calculated intracellular pyruvate concentrations, the following conclusions were drawn with respect to product formation of cells growing under glucose limitation. (i) Pyruvate decarboxylase, the key enzyme of alcoholic fermentation, probably already is operative under conditions in which alcoholic fermentation is absent. The acetaldehyde produced by the enzyme is then oxidized via acetaldehyde dehydrogenases and acetyl-CoA synthetase. The acetyl-CoA thus formed is further oxidized in the mitochondria. (ii) Acetate formation results from insufficient activity of acetyl-CoA synthetase, required for the complete oxidation of acetate. Ethanol formation results from insufficient activity of acetaldehyde dehydrogenases.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2566301 TI - T4+ cell production of interferon gamma and the clinical spectrum of patients at risk for and with acquired immunodeficiency syndrome. PMID- 2566303 TI - Treatment of cancer pain with orally and spinally administered opioids. PMID- 2566302 TI - Genome type analysis of adenoviruses: isolates from one year from the Hannover area. AB - Adenoviruses (AV), isolated from 138 children during the year 1981 in the Hannover area, were studied by DNA restriction analysis with the enzymes BamHI, BglII, BstEII, EcoRI, HindIII, KpnI, and SmaI and compared with the respective prototypes. Varying fragment patterns were depicted and genome types analyzed. Prototype-like strains of AV1 and 5 were not found. Types 2, 5, 1, 3, and 7 showed decreasing genetic variation in that order. Altered restriction sites were physically mapped on the genome; they appeared to be randomly distributed. The high genetic variability of AV2 and 5 is remarkable for this study population, limited in time and space. Clinical and epidemiological data were also presented in relation to serotypes. PMID- 2566304 TI - Homeobox genes and development of the vertebrate CNS. AB - The discovery of homeobox genes in vertebrates may allow analysis of a basic problem in developmental neurobiology: how regional differences in CNS organization are specified during development. This view is based on the roles defined for homologous genes in Drosophila development, and is supported by studies of the patterns of homeobox gene expression in vertebrate embryos. Homeobox genes comprise a multigene family, members of which are expressed in different spatially restricted domains along the anterior-posterior axis of the CNS. These observations are consistent with homeobox genes having roles in the positional specification of CNS organization, and experimental tests of this should be forthcoming shortly. PMID- 2566305 TI - Mechanisms underlying induction and maintenance of long-term potentiation in the hippocampus. AB - Long-term potentiation (LTP) in the hippocampus is accompanied by a number of changes on both sides of the synapse. It is now generally considered that the trigger for initiating LTP is the entry of calcium into the postsynaptic area through the NMDA-associated channel while the mechanism(s) underlying the maintenance of LTP are less well understood and probably involve contributions from both sides of the synapse. PMID- 2566306 TI - Characterization of a somatostatin-28 generating metallo-endoprotease from rat brain cytosol. AB - Brain cytosol contains a neutral metallo-protease of about 80,000 which cleaves a substrate containing the site at which mammalian prosomatostatin is cleaved to generate somatostatin 28 in vivo. This represents a cleavage on the carboxyl side of a single arginine residue at an Arg-Ser bond. The enzyme was unable to cleave several other substrates containing single arginine residues or two substrates containing an Arg-Lys or Lys-Arg pair. When it was incubated with anglerfish pancreatic prosomatostatin, it produced significant quantities of a peptide which co-eluted with somatostatin 28 II. Based on the ability of this enzyme to cleave small and large substrates related to somatostatin, it is a potential candidate for the enzymes which cleaves prosomatostatin in vivo. PMID- 2566307 TI - Selective potentiation by L-cysteine of apparent binding activity of [3H]glutathione in synaptic membranes of rat brain. AB - Significant apparent binding activity of [3H]glutathione was detected in synaptic membranous preparations of the rat brain. In vitro addition of sucrose (50-1000 mM) and Triton X-100 (0.02-0.1%) significantly diminished the apparent binding activity, whereas pretreatment of the membranes with Triton X-100 (0.01-0.4%) did not affect the activity. A slight but statistically significant reduction of the apparent binding activity was induced by the in vitro addition (1 mM) of two constituent amino acids, L-glutamic acid and glycine. In contrast, another constituent amino acid, L-cysteine, potently enhanced the binding activity at a concentration higher than 0.1 mM. No prominent alteration of the activity occurred following the inclusion of structurally-related amino acids, dithiothreitol, dithioerythritol and numerous other amino acids. Scatchard analysis revealed that the apparent binding consisted of two independent separate components with Kd values of 0.76 and 11.0 microM, and Bmax values of 4.00 and 27.0 pmol/mg protein respectively. In vitro addition of 1 mM L-cysteine resulted in a single component with a Kd of 8.5 microM and a Bmax of 105 pmol/mg protein. Pretreatment of the membranes with 1 mM L-cysteine potentiated the apparent binding, with a further addition of L-cysteine having no effect. The retina had the highest activity followed by the hypothalamus, striatum, spinal cord, midbrain, hippocampus, medulla-pons, cerebellum and cerebral cortex, which occurred independently of the incubation temperature. In peripheral organs examined, the pituitary possessed higher activity than the retina, with progressively lower activities in the adrenal, liver, spleen, skeletal muscle and heart. No significant activity was detected in the kidney. Addition of 1 mM L cysteine significantly potentiated the activities at 30 degrees, but not at 2 degrees, in the hippocampus and cerebral cortex without affecting those in other central structures. In contrast, a profound inhibition of the activity was induced by the addition of L-cysteine in the pituitary, adrenal, intestinal mucosa, skeletal muscle and retina independently of the temperature. These results suggest that L-cysteine may selectively potentiate the apparent binding activity of [3H]glutathione in particular regions of the brain, while eliminating that in the peripheral excitable tissues. PMID- 2566308 TI - Cardiovascular effects of the new cardiotonic agent 1,2-dihydro-6-methyl-2-oxo-5 (imidazo[1,2-a]pyridin-6-yl)-3-pyridine carbonitrile hydrochloride monohydrate. 1st communication: studies on isolated guinea pig cardiac muscles. AB - Cardiotonic effects and the mechanism of action of 1,2-dihydro-6-methyl-2-oxo-5 (imidazo[1,2-a]pyridin-6-yl)-3-pyridine carbonitrile hydrochloride monohydrate (E 1020), a new cardiotonic agent, were investigated in vitro. E-1020 (10(-7)-10(-4) mol/l) produced a concentration-dependent positive inotropic effect in papillary muscles. E-1020 also caused an increase in contractile force in the right atria which was accompanied by small increases in spontaneous beating rate. The inotropic effect of E-1020 on papillary muscles was not altered by treatment with beta-adrenoceptor or histamine H2-receptor blockade, but was attenuated by the muscarinic agonist, carbachol. E-1020, like isoprenaline (isoproterenol, Iso), restored the contraction of papillary muscles which had been arrested in a high potassium solution. The inotropic response of papillary muscles to Iso was potentiated by pretreatment with E-1020 at a minimally-effective inotropic concentration (3 x 10(-7) mol/l). E-1020 did not affect the activity of dog kidney Na+, K+-ATPase. On the other hand, the compound specifically inhibited the cyclic AMP-specific isoenzyme (fraction III) of phosphodiesterase and caused an elevation of cyclic AMP content in guinea pig hearts. These results indicate that E-1020 is a potent cardiotonic agent with a minor chronotropic effect and that its inotropic action is mainly mediated by the rise in cardiac cyclic AMP content due to the inhibition of cyclic AMP-specific phosphodiesterase. PMID- 2566309 TI - Plasma lipoprotein lipids in relation to the MspI polymorphism of the apolipoprotein AII gene in Caucasian men. Lack of association with plasma triglyceride concentration. AB - Digestion of the human apolipoprotein (apo) A-II gene with the endonuclease MspI produces fragments of 3.0 or 3.7 kb, reflecting the presence or absence of a polymorphic site within an Alu sequence 3' to the gene. Patients with hypertriglyceridemia have been shown to have an increased prevalence of the 3.0 kb allele. To explore this observation further, plasma lipoprotein lipids were studied in a random sample of fasted middle-aged Caucasian men, of which 59 were 3.0 kb homozygotes, 24 were heterozygotes, and 2 were 3.7 kb homozygotes. After adjusting for the effects of age, height, weight, alcohol intake and cigarette consumption by covariance analysis, no statistically significant associations were present between genotype and the concentrations of triglyceride in whole plasma or the d less than 1.019 g/ml fraction of plasma (i.e., VLDL + IDL). Nor were the cholesterol concentrations in VLDL + IDL, low density lipoprotein (LDL, d = 1.019-1.063 g/ml), high density lipoprotein (HDL), HDL2 or HDL3 related to genotype. In an independent comparison of eight 3.0 kb homozygotes and eight 3.7 kb homozygotes (all Caucasians) drawn from a different community, genotype was unrelated to the triglyceride or cholesterol concentrations in VLDL (d less than 1.006 g/ml), IDL + LDL (d = 1.006-1.063 g/ml) or HDL, after adjustment for the effects of covariates. These results suggest that the MspI polymorphism of the apo A-II gene is not associated with genetic variation that significantly affects triglyceride transport in the majority of men.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566310 TI - Urinary enzyme output during detoxification of chronic alcoholic patients. AB - The urinary excretion of N-acetylglucosaminidase, gamma-glutamyl transferase and lactate dehydrogenase was elevated by two to three fold in chronic alcoholic patients when compared to normal controls. After 4 weeks of alcohol abstention, the urinary excretion of these enzymes returned to control values. Increased urinary enzyme excretion was not related to malnutrition or pathological glomerular filtration and is due most probably to a direct effect of alcohol on the renal proximal tubule. PMID- 2566311 TI - Acetaldehyde-induced alterations in metabolism of monoamines in mouse brain. AB - The effects of single and repeated inhalations of acetaldehyde (AcAl) on spontaneous activity and the metabolisms of cerebral monoamines and neuroactive amino acids were investigated. Both single and repeated inhalations of AcAl induced a significant increase of spontaneous activity at the initial stage followed by the loss of motor activity and coma. The AcAl inhalation-induced central excitement, exhibited by hyperkinesia and occasional jumpings, were found to be more severe following a single administration than by repeated ones. These abnormal behaviors observed following a single administration of AcAl, were accompanied by significant decreases of noradrenaline in the cerebral cortex, brainstem and of dopamine in the brainstem. In addition, it was found that these decreases in catecholamines were associated with significant decreases in the contents of 3-methoxy-4-hydroxyphenylethyleneglycol, 3, 4-dihydroxyphenylacetic acid and homovanillic acid in brain. The contents of aspartic acid in the cerebral cortex and of GABA in the brainstem also showed an increase. On the other hand, animals subjected to the repeated inhalation of AcAl and exhibiting a state of central depression, showed the increase of adrenaline as well as the decrease of GABA in the cerebellum. These results suggest that a single inhalation of AcAl may induce central excitation as well as facilitate the metabolic turnover of cerebral catecholamines, while repeated inhalation of AcAl may result in central depression accompanied by decreased turnover of central catecholamines. Possible involvement of the changes in cerebral aspartic acid and GABA in the exhibition of central effects of AcAl is also suggested. PMID- 2566312 TI - Inhibition of the brain to blood transport system for enkephalins and Tyr-MIF-1 in mice addicted or genetically predisposed to drinking ethanol. AB - Enkephalin concentrations in the brain correlate inversely with ethanol intake and the predisposition of different strains of mice to drink. This and other evidence link ethanol ingestion, addiction, and withdrawal to opiate peptides. We studied the effect of these conditions on the saturable, stereospecific system that transports the enkephalins and Tyr-MIF-1 (a peptide with antiopiate action) out of the brain. The transport rate in mice physically dependent on ethanol was only 56% of the rate in control mice, but during withdrawal from ethanol transport rates increased to levels seen in controls. Transport rates were also lower in strains of mice previously determined to have lower enkephalin concentrations in the brain and to be predisposed to drinking ethanol. Acute intraperitoneal or intracerebroventricular injection of ethanol had minimal direct effects on the transport rate, suggesting that it is not ethanol itself, but those factors associated with addiction and the predisposition to drink ethanol, that altered transport. These studies raise the possibility that the inhibition of this system that transports enkephalins/Tyr-MIF-1 out of the brain might offer a new approach to the control of drinking and withdrawal from ethanol. PMID- 2566313 TI - Antagonism of selected ethanol-enhanced brain stimulation properties by Ro 15 4513. AB - Low doses of ethanol increase responding for brain stimulation. Recently, other intoxicating effects of ethanol have been reversed by the imidazobenzodiazepine, Ro 15-4513. Possibly, Ro 15-4513 blockade also acts on reward-enhancing properties of ethanol. Rats trained to alternately shuttle between nose poke and lever operanda for rewarding stimulation to the medial forebrain bundle, were tested following intragastric intubations of ethanol (18%, 1.35 g/kg), Ro 15-4513 (3 mg/kg in 18% ethanol), or vehicle. Ro 15-4513 reversed ethanol-enhanced effects on reinforced responses. Because Ro 15-4513 did not completely block instrumental responding for brain stimulation, we conclude its effects on ethanol were not acting on the same reward substrate as the current and consummatory response. PMID- 2566314 TI - Regulation of angiotensinogen gene. AB - The development of recombinant DNA technology has introduced new directions for the study of the angiotensinogen molecule. The cloning and sequencing of the human and rat cDNAs demonstrate the similarity of angiotensinogen to various serine protease inhibitors produced by the liver and was the beginning of studies looking for new physiological roles of angiotensinogen, in addition to the substrate for renin. The determination of the nucleotide sequence of these cDNAs also allowed the identification of angiotensinogen mRNA in many tissues in addition to the liver that is the major site of synthesis. This multilocalization of angiotensinogen is one of the arguments for the presence and the function of local renin-angiotensin systems. Finally, the hepatic biosynthesis of angiotensinogen is regulated by many different hormonal factors including glucocorticoid, estrogen, thyroid hormone, insulin, and angiotensin II. The cloning of the angiotensinogen gene offers the opportunity to study this regulation at the transcriptional level. We present in this paper a review of the literature concerning the new aspects of angiotensinogen using molecular biological tools and its regulation together with the characterization of the human angiotensinogen gene. PMID- 2566315 TI - Maladaptive behaviors associated with neuroleptic drug maintenance. AB - In this study we examined whether the decision to maintain mentally retarded persons on neuroleptic drugs was associated with specific maladaptive behaviors. A drug-withdrawal group, a drug-reinstatement group, and a drug-control group were used. A two-way repeated measures multivariate analysis of variance design indicated that mentally retarded persons were more likely to be maintained on drugs because of events that were perceived as significant and recorded by institutional staff in clinical notes than because of the reasons for which the drugs were prescribed initially. In particular, staff members' perceptions of disturbing behavior, self-stimulation, and physical aggression were likely to influence their decision to maintain mentally retarded persons on neuroleptics. PMID- 2566317 TI - Liver cell necrosis and regeneration following injections of carbon tetrachloride. Effects of the thyrotropin-releasing hormone and somatostatin. AB - Mice were given 10 micrograms somatostatin or 25 micrograms TRH intraperitoneally 10 min before s.c. injection of 2 or 20 mg CCl4. The extent of liver cell necrosis and nuclear size were measured by the electronic Mini Mop method and the extent of necrosis and nuclear pleomorphism were estimated by a visual linear analogue scale of 100 mm, and compared to plasma concentrations of ASAT and ALAT. Pre-treatment with TRH or somatostatin resulted in significant reduction in the extent of necrosis 24 h after CCl4-injections (25%), with a lowering of ASAT from 13209 +/- 2955 U/l to 5144 +/- 924 after TRH and to 6186 +/- 966 after somatostatin, and of ALAT from 14343 +/- 3209 to 7718 +/- 1727 and 6494 +/- 1253 U/l, respectively. After 3 days the necroses were reduced from 16.5 +/- 1.7% by the Minimop method to 1.4 +/- 0.5% (90%) in mice given CCl4 alone, and from 12.3 +/- 1.7% to 3.8 +/- 1.2% in mice pretreated with TRH, and from 12.3 +/- 1.8% to 3.8 +/- 1.7% (70%) in mice pretreated with somatostatin. The plasma concentrations of ASAT and ALAT were reduced correspondingly. After 5 days no necroses were seen, and the plasma ASAT and ALAT were normal. After 6 months of weekly injections of TRH or somatostatin before 20 mg CCl4 the liver cell nuclear size (10.5 and 9.7 0.3 mu 2) was similar to that after CCl4 alone (9.7 0.3 mu 2), and twice that of controls (4.6-5.4 0.1 mu 2). Liver cell necrosis was not seen. The plasma concentrations of ASAT (131 8.6-162 11.3) and ALAT (98 8-104 9 Iu/l) were similarly 2-3 times those in controls. TRH and somatostatin thus reduced liver cell injury and delayed regeneration after single injections of CCl4. After 6 months of weekly injections no effects were observed. PMID- 2566318 TI - Guanine ribonucleotide metabolism in human red blood cells: evidence for a high rate of GMP dephosphorylation. AB - The flux rates through the metabolic pathways affecting the maintenance of GuRN pool in intact human RBC were studied. Normal RBC, incubated in KRBB, exhibited a markedly higher accumulation in nucleotides of Gu than of Hx. Addition of 8-AGuo, a potent inhibitor of PNP, resulted in a marked increase in the accumulation of label in the nucleosides, in Ino following incubation with Hx, and in Guo following incubation with Gu, indicating a very high rate of IMP and GMP degradation to bases through their respective nucleosides. Most of the degradation of GMP is by dephosphorylation to Guo, rather than through reductive deamination to IMP. The ultimate fate of IMP in RBC is its degradation to Ino and consequently to Hx. The contribution of AdRN or of IMP to the GuRN pool is negligible. The results indicate that concerning IMP and GMP, human RBC contain very active futile cycles, nucleotide----nucleoside----base----nucleotide, catalyzed by 5'-nucleotidase, PNP, and HGPRT. The operation of the complete cycles is essential for the maintenance of GuRN and the IMP pool size. These results may explain the finding of reduced GTP content in RBC from patients with an inborn deficiency of PNP or of HGPRT. PMID- 2566319 TI - Induction of expression of HIV in latently or chronically infected cells. PMID- 2566316 TI - Psychotropic medication of mentally retarded residents in community long-term care facilities. AB - Medicaid billing information was used to examine the administration of psychotropic medication to residents of community long-term care facilities providing mental retardation services. In 1984, 5,766 individuals receiving Medicaid in Illinois resided continuously in these facilities for the entire year. Of these, 1,667 (28.9%) received at least one psychotropic medication during the year, with thioridazine, haloperidol, and chlorpromazine being prescribed most frequently. Data for frequency, dose, and length of administration of individual psychotropic medications were reported for the total group and by level of mental retardation. Separate analyses indicated little or no influence of demographic and facility variables on either the probability of drug administration or amount administered. PMID- 2566320 TI - Expression of HIV antigens at the surface of infected T4 cells: immunoelectron microscopic evidence of an immunogenic phase prior to the viral release. AB - HIV antigens were detected by immunoelectron microscopy at the surface of human and simian T4 lymphocytes that had been infected in vitro. HIV antigens were detected at the surface of cells exhibiting viral particles but also at the surface of cells before the release of virions. The latter cells may be considered immunogenic since they are capable of triggering specific immune responses without the cytopathic effects due to viral release. PMID- 2566321 TI - Comparison of the cardiovascular and pulmonary effects of oral celiprolol, propranolol and placebo in normal volunteers. AB - 1. The effects on heart rate, blood pressure and pulmonary function of single oral doses of celiprolol hydrochloride (400 mg), and propranolol (40 mg) were compared with placebo in 12 healthy volunteers, in a double-blind three-period crossover study. 2. Celiprolol had no effect on heart rate while propranolol caused a significant reduction compared with placebo. Systolic blood pressure was reduced by propranolol but not celiprolol, whereas standing diastolic blood pressure was lowered by both drugs. 3. The maximal expiratory flow at 50% vital capacity (MEF.50), was significantly lower after propranolol compared with placebo and celiprolol. Celiprolol had no effect on the flow-volume loop parameters. 4. Effective pulmonary blood flow was significantly increased by celiprolol, but reduced by propranolol. 5. A high incidence of subjective side effects were experienced on celiprolol (10/12; particularly unpleasant in 5). Side-effects were experienced to a lesser extent on placebo (8/12). Only one volunteer experienced a side-effect on propranolol. 6. Oral celiprolol exerts its hypotensive effect by vasodilatation without reflex tachycardia. It does not cause airways obstruction in healthy subjects. PMID- 2566322 TI - Kinetic and magnetic resonance studies of the glutamate-43 to serine mutant of staphylococcal nuclease. AB - The Glu-43 residue of staphylococcal nuclease has been proposed to function as a general base that facilitates the attack of water on the phosphodiester substrate [Cotton, F. A., Hazen, E. E., & Legg, M. J. (1979) Proc. Natl. Acad. Sci. U.S.A. 76, 2551-2555]. With DNA as substrate, Vmax in the glutamate-43--serine (E43S) mutant enzyme is decreased by 2700-fold at pH 7.4 but only 376-fold at pH 9.9. With the wild-type enzyme, Vmax increases with pH to pH 9.2, above which it becomes less sensitive to further increase in pH, leveling off at pH 9.8. In contrast, Vmax of the E43S mutant continues to rise, first order in [OH-], to pH 9.8. Above pH 10 both activities fall irreversible. Hence the hydroxyl ion can partially replace the effect of Glu-43 on kcat, in accord with the proposed role of Glu-43 as a general base. The inflection point in the curve relating pH to log Vmax of the wild-type enzyme at pH 9.4 may reflect the ionization of a Ca2+-bound water, or of a Lys or Tyr residue at the active site. The activator Ca2+ and the competitive inhibitor Mn2+ bind to the E43S mutant an order of magnitude more weakly than to the wild-type enzyme as detected by kinetics and by direct metal binding studies, and approximately one additional water ligand on Mn2+ is found in the binary Mn2+ complex of the E43S mutant (1.4 +/- 0.2) as compared to that of the wild-type enzyme (0.8 +/- 0.2). These data suggest that Glu-43 coordinates the divalent cation in the binary enzyme-metal complex but dissociates from the metal to create a water binding site and to function as a general base in the ternary enzyme-metal-DNA complex. While a 2-fold weaker binding of DNA to the Ca2+ complex of the E43S mutant than to the wild-type enzyme is found by kinetic studies, an order of magnitude tighter binding of the competitive inhibitor 3',5' pdTp to the Mn2+ and Ca2+ complexes of E43S is found by direct binding studies. Distances from Co2+ to phosphorus in the ternary enzyme-Co2+-pdTp complexes reveal coordination of only the 5'-phosphate by Co2+ on the wild-type enzyme but coordination of both the 3'- and 5'-phosphates of pdTp on the E43S mutant.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2566323 TI - 1H NMR study of the solution properties of the polypeptide neurotoxin I from the sea anemone Stichodactyla helianthus. AB - The solution properties of the polypeptide neurotoxin I from the sea anemone Stichodactyla helianthus (Sh I) have been investigated by high-resolution 1H nuclear magnetic resonance (NMR) spectroscopy at 300 MHz. The pH dependence of the spectra has been examined over the range 1.1-12.2 at 27 degrees C. Individual pKa values have been obtained for the alpha-ammonium group of Ala-1 (8.6) and the side chains of Glu-8 (3.7), Tyr-36 (10.9), and Tyr-37 (10.8). For the remaining seven carboxyl groups in the molecule (from five Asp, Glu-31, and the C terminus), four pKa values, viz., 2.8, 3.5, 4.1 and 6.4, can be clearly identified. The five Lys residues titrate in the range 10.5-11, but individual pKa values could not be obtained because of peak overlap. Conformational changes associated with the protonation of carboxylates occur below pH 4, while in the alkaline pH range major unfolding occurs above pH 10. The molecule also unfolds at elevated temperatures, having a transition temperature of ca. 55 degrees C at pH 5.25. Exchange of the backbone amide protons has been monitored at various values of pH and temperature in the ranges pH 4-5 and 12-27 degrees C. Up to 18 slowly exchanging amides are observed, consistent with the existence of a core of hydrogen-bonded secondary structure, most probably beta-sheet.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566324 TI - Sequence-specific 1H NMR assignments and secondary structure in the sea anemone polypeptide Stichodactyla helianthus neurotoxin I. AB - Sequence-specific assignments are reported for the 500-MHz 1H nuclear magnetic resonance (NMR) spectrum of the 48-residue polypeptide neurotoxin I from the sea anemone Stichodactyla helianthus (Sh I). Spin systems were first identified by using two-dimensional relayed or multiple quantum filtered correlation spectroscopy, double quantum spectroscopy, and spin lock experiments. Specific resonance assignments were then obtained from nuclear Overhauser enhancement (NOE) connectivities between protons from residues adjacent in the amino acid sequence. Of a total of 265 potentially observable resonances, 248 (i.e., 94%) were assigned, arising from 39 completely and 9 partially assigned amino acid spin systems. The secondary structure of Sh I was defined on the basis of the pattern of sequential NOE connectivities, NOEs between protons on separate strands of the polypeptide backbone, and backbone amide exchange rates. Sh I contains a four-stranded antiparallel beta-sheet encompassing residues 1-5, 16 24, 30-33, and 40-46, with a beta-bulge at residues 17 and 18 and a reverse turn, probably a type II beta-turn, involving residues 27-30. No evidence of alpha helical structure was found. PMID- 2566325 TI - A proton nuclear magnetic resonance study of the antihypertensive and antiviral protein BDS-I from the sea anemone Anemonia sulcata: sequential and stereospecific resonance assignment and secondary structure. AB - The sequential resonance assignment of the 1H NMR spectrum of the antihypertensive and antiviral protein BDS-I from the sea anemone Anemonia sulcata is presented. This is carried out with two-dimensional NMR techniques to identify through-bond and through-space (less than 5 A) connectivities. Added spectral complexity arises from the fact that the sample is an approximately 1:1 mixture of two BDS-I isoproteins, (Leu-18)-BDS-I and (Phe-18)-BDS-I. Complete assignments, however, are obtained, largely due to the increased resolution and sensitivity afforded at 600 MHz. In addition, the stereospecific assignment of a large number of beta-methylene protons is achieved from an analysis of the pattern of 3J alpha beta coupling constants and the relative magnitudes of intraresidue NOEs involving the NH, C alpha H, and C beta H protons. Regular secondary structure elements are deduced from a qualitative interpretation of the nuclear Overhauser enhancement, 3JHN alpha coupling constant, and amide NH exchange data. A triple-stranded antiparallel beta-sheet is found to be related to that found in partially homologous sea anemone polypeptide toxins. PMID- 2566326 TI - Determination of the three-dimensional solution structure of the antihypertensive and antiviral protein BDS-I from the sea anemone Anemonia sulcata: a study using nuclear magnetic resonance and hybrid distance geometry-dynamical simulated annealing. AB - The three-dimensional solution structure of the antihypertensive and antiviral protein BDS-I from the sea anemone Anemonia sulcata has been determined on the basis of 489 interproton and 24 hydrogen-bonding distance restraints supplemented by 23 phi backbone and 21 chi 1 side-chain torsion angle restraints derived from nuclear magnetic resonance (NMR) measurements. A total of 42 structures is calculated by a hybrid metric matrix distance geometry-dynamical simulated annealing approach. Both the backbone and side-chain atom positions are well defined. The average atomic rms difference between the 42 individual SA structures and the mean structure obtained by averaging their coordinates is 0.67 +/- 0.12 A for the backbone atoms and 0.90 +/- 0.17 A for all atoms. The core of the protein is formed by a triple-stranded antiparallel beta-sheet composed of residues 14-16 (strand 1), 30-34 (strand 2), and 37-41 (strand 3) with an additional mini-antiparallel beta-sheet at the N-terminus (residues 6-9). The first and second strands of the triple-stranded antiparallel beta-sheet are connected by a long exposed loop (residues 17-30). A number of side-chain interactions are discussed in light of the structure. PMID- 2566327 TI - NMR studies of toxin III from the sea anemone Radianthus paumotensis and comparison of its secondary structure with related toxins. AB - Nearly complete assignments of the proton nuclear magnetic resonance (NMR) spectrum of the polypeptide toxin III from the sea anemone Radianthus paumotensis (RP) are presented. The secondary structures of the related toxins RP II and RP III are described and are compared with each other and with another related toxin ATX Ia from Anemonia sulcata [Widmer, H., Wagner, G., Schweitz, H., Lazdunski, M., & Wuthrich, K. (1988) Eur. J. Biochem. 171, 177-192]. All of these proteins contain a highly twisted four-strand antiparallel beta-sheet core connected by loops of irregular structure. From the work done with AP-A from Anthopleura xanthogrammica [Gooley, P. R., & Norton, R. S. (1986) Biochemistry 25, 2349 2356], it is clear that this homologous toxin also has the same basic core. Some small differences are seen in the structures of these toxins, particularly in the position of the N-terminal residues that form one of the outside strands of the beta-sheet. In addition, the R. paumotensis toxins are two residues longer, extending the third strand of sheet containing the C-terminal residues. A comparison of chemical shifts for assigned residues is also presented, in general supporting the similarity of structure among these proteins. PMID- 2566328 TI - Determination of amino- and carboxyl-terminal sequences of guinea pig liver transglutaminase: evidence for amino-terminal processing. AB - Transglutaminases (EC 2.3.2.13) catalyze the formation of epsilon-(gamma glutamyl)lysine cross-links and the substitution of a variety of primary amines for the gamma-carboxamide groups of protein-bound glutaminyl residues. These enzymes are involved in many biological phenomena. In this study, the amino- and carboxyl-terminal sequences of guinea pig liver transglutaminase were identified by sequence analysis to determine whether this enzyme is processed posttranslationally at its terminal regions. Two peptides, believed to contain the amino-terminal sequences of transglutaminase, were isolated from the Pronase digest of the enzyme protein with SP-Sephadex C-25 column chromatography and reverse-phase HPLC. Analyses (amino acid analysis, sequencing after the treatment with an acylamino-acid-releasing enzyme, and fast atom bombardment mass spectrometry) of these peptides indicated that the amino-terminal structure of this enzyme is acetylAla-Glu-Asp-Leu-Ile-Leu-Glu. The candidate for the carboxyl terminal peptide in the trypsin digest of enzyme was isolated from the unadsorbed fraction of affinity chromatography with anhydrotrypsin agarose gel. The peptide was found to be Asn-Val-Ile-Ile-Gly-Pro-Ala. Both the terminal sequences were completely consistent with those predicted from the cDNA sequence [Ikura, K., Nasu, T., Yokota, H., Tsuchiya, Y., Sasaki, R., & Chiba, H. (1988) Biochemistry 27, 2898-2905]. These results indicated that the amino-terminal processing occurred after or in the course of translation of this enzyme, namely, removal of the initiator methionine and a subsequent acetylation of the alanine residue adjacent to the methionine. Our results did not indicate carboxyl-terminal processing of guinea pig liver transglutaminase. PMID- 2566329 TI - Differential inhibitory action of the fungal toxin orellanine on alkaline phosphatase isoenzymes. AB - The inhibitory action of orellanine (3,3',4,4'-tetrahydroxy-2,2'-dipyridyl-1,1' dioxide), a fungal toxin of Cortinarius orellanus Fr. and C. orellanoides R. Hry., on alkaline phosphatase isoenzymes was studied. Orellanine specifically inhibited alkaline phosphatase activity in LLC-PK1 renal epithelial cell cultures and in the colon carcinoma cell line Caco-2 without affecting gamma-glutamyl transpeptidase activity. Kinetic studies revealed that orellanine acts on renal alkaline phosphatase as a noncompetitive inhibitor, whereas the intestinal and placental isoforms are inhibited competitively. PMID- 2566330 TI - [Mapping of the thyroglobulin gene in high molecular weight DNA from the human thyroid in normal conditions and in thyroid pathology]. AB - A highly purified thyroglobulin (Tg) mRNA was isolated from human nodal euthyroid goiter. Ultracentrifugation in a 5-20% sucrose density gradient revealed that the protein has a sedimentation coefficient of 33S. cDNA was synthesized from the 33S RNA, using reverse transcriptase in the presence of a human placental ribonuclease inhibitor. In order to detect the polymorphism of restriction fragment length, screening of high molecular weight DNA from normal human thyroid gland and from nodal euthyroid and diffuse toxic goiters was carried out, using Tg cDNA probes and plasmid DNA containing a Tg cDNA fragment. After restriction with endonuclease Hind III, three Tg-containing fragments were identified both in the normal and nodal euthyroid goiters, whereas restriction with endonuclease Bam HI revealed two Tg-containing fragments in nodal euthyroid and diffuse toxic goiters. PMID- 2566332 TI - Maternal and fetal endocrine pancreas in the spontaneously diabetic BB rat. AB - The maternal and fetal endocrine pancreas were investigated in the diabetic BB rat on day 21 of pregnancy. The maternal pancreas of the diabetic rat contained practically no insulin-positive B cells. The A and D cell mass were also decreased, while plasma glucagon and somatostatin levels were normal or increased, confirming previous data. Six of 11 diabetic rats had B-cell-specific surface antibodies (ICSA), whereas only 1 of 10 nondiabetic rats was ICSA positive. The volume density of insulin-positive cells was decreased in the fetal pancreas of diabetic BB rats compared to fetuses of nondiabetic rats, but the volume density of glucagon- and somatostatin-positive cells remained normal. The B cells of these fetuses were ultrastructurally less granulated and showed signs of increased cellular activity. Plasma insulin levels were decreased while plasma glucagon and somatostatin concentrations were normal. ICSA were not detected in fetuses of nondiabetic and diabetic rats. There were no differences in the histology of the spleen and thymus between both groups of fetuses. Metabolic characterization of the growth-retarded fetuses of diabetic rats revealed, besides lower plasma insulin concentrations, increased branched chain amino acid levels, and normal plasma Sm/IGF-I levels. The main conclusions from this study are: (1) Severe maternal diabetes decreases the pancreatic insulin-positive cell mass and plasma insulin levels in the fetus, but not the A and D cell mass and function; (2) ICSA are not detectable in fetal plasma; (3) the influence of maternal BB rat diabetes on fetal endocrine pancreas and metabolic environment resembles that of severe streptozotocin-induced diabetes. PMID- 2566331 TI - Low intestinal lactase activity in offspring from ethanol-treated mothers. AB - Some aspects of small intestine maturation have been studied in the newborns from chronic ethanol-treated pregnant rats (25% ethanol in drinking fluid) immediately after birth (before suckling) and after 30 days of life. Litters delivered by mothers fed ad libitum with a standard diet diluted 50% with cellulose were used as a nutritional control. At birth, pups from ethanol-treated mothers showed significant decreases in total intestinal length and thickness, low total lactase activity and low somatostatin intestinal content. The intestinal alterations of these neonatal parameters are not present in newborns from mothers on fiber diluted diet. From delivery, pups from different experimental groups were nursed by normal lactating dams. At 30 days of age neither of those parameters differed among the groups. We propose that the low levels of total lactase activity in newborns from alcoholic mothers, that are a consequence of a lower intestinal mucosa content, are a direct effect of ethanol in utero on the fetal gastrointestinal system. PMID- 2566333 TI - Anomalous dominance and persistent tardive dyskinesia. AB - We examined handedness and cerebral hemispheric asymmetries on computed tomography (CT) scan in a sample of schizophrenic patients who were rated also for the presence or absence of persistent tardive dyskinesia (TD). Patients with TD showed a more standard dominance pattern, with dextral hand preference and normal occipital asymmetry. Anomalous dominance was associated with a marked underrepresentation of TD. Stepwise discriminant analyses indicated that the statistical prediction of TD was enhanced by the inclusion of dominance measures. Schizophrenic patients with strong standard dominance patterns may be more susceptible to developing TD, or conversely, anomalous dominance may confer protection against TD. PMID- 2566334 TI - Neuroleptics reverse attention asymmetries in schizophrenic patients. AB - Discussions of hemispheric asymmetry in psychopathology are often confounded by the effects of medication. We examined the effect of neuroleptic drugs on attention asymmetries in acutely psychotic patients admitted for the first time to a psychiatric hospital before the initiation of drug treatment and again after a period of treatment with neuroleptics. Overall performance did not change significantly; however, attention asymmetry was clearly related to the medication status of the patient: unmedicated patients showed inattention to the right hemispace, which changed to more prominent left-sided inattention when medicated. A longer time on medication or a higher daily dose were associated with a shift of inattention from the right to left hemispace. This suggests that neuroleptics may normalize left hemisphere performance, at the expense of deteriorated right hemisphere performance. PMID- 2566335 TI - Are disturbances in lipid-protein interactions by phospholipase-A2 a predisposing factor in affective illness? AB - Current theories of affective disorders do not account for many of the biological markers replicated in patient studies. We link many biological findings in a reasonable physiological relationship, compatible with mechanisms of action of pharmacological and electroshock therapies for depression. We propose that excessive phospholipase-A2 (PLA2) activity disrupts membrane fluidity, composition, and therefore, the activity, of membrane-dependent proteins. Similar disruptions in these proteins are documented in depressed patients and can be accounted for by excessive PLA2 activity. This paradigm accounts for disturbances in the activity of Na-K-ATPase, beta2- and alpha2-adrenergic receptors, MAO, norepinephrine and serotonin uptake, and imipramine binding. Disturbances in other membrane-dependent proteins, tyrosine and tryptophan hydroxylase, can explain the biogenic amine hypothesis. Inhibition of glucocorticoid receptor and TRH receptor binding to their respective ligands by PLA2 may explain patient nonsuppression in the Dexamethasone Suppression Test and poor response in the TRH stimulation test. Physiological regulators of PLA2 activity; calcium, cortisol, estrogen, progesterone, and PGE2 are documented abnormalities in some patients with affective disorders and consistent with excessive PLA2 activity. Thus, postpartum depression and premenstrual tension syndrome may be described in the paradigm. The mechanisms of action of tricyclic antidepressants, lithium, electroconvulsive shock, and some novel antimanic agents can be described in terms of alterations of PLA2 activity. Interestingly, ethanol perturbs membrane fluidity and membrane-bound enzymes in a manner similar to excessive PLA2 activity. A hereditary factor predisposing patients to affective disorders may be a gene defect at either PLA2 or in its regulation. PMID- 2566337 TI - Single oral dose proportionality pharmacokinetics of almitrine bismesylate in humans. AB - A single-blind study was conducted in 10 healthy male subjects. Each subject was tested with four single oral doses of capsules containing 25, 50, 100, 200mg almitrine bismesylate and one dose of placebo. Blood samples were drawn as a function of time and the concentration of almitrine in plasma was determined by gas chromatography utilizing nitrogen-phosphorus detection. Linear regression analysis of the data suggested that a deviation from linearity existed between the area under the plasma concentration time curves and the dose (R = 0.96). Linear analysis of the individual data indicates that a slight negative deviation from linearity is apparent for the 200 mg dose. The same trend was observed for the mean maximum almitrine plasma concentration, Cmax, which ranged from 38.9 +/- 11.8 to 286.2 +/- 99.1 ng ml-1 for the 25 and 200 mg dose, respectively. The time to peak was relatively constant regardless of the administered dose and ranged from 2.4 +/- 0.5 h to 2.8 +/- 0.8 h. Good agreement was obtained between the observed bioavailability parameters and those predicted from the nonlinear fit of the data. Further kinetic analysis of the data revealed mean total body clearance over fraction of dose absorbed ranging from 268.2 +/- 132.8 to 436.4 +/- 191.4 ml min-1 for doses 50 and 200mg, respectively. PMID- 2566336 TI - Effect of Ca2+ diffusion on the time course of neurotransmitter release. AB - The three-dimensional (3D) diffusion model of Fogelson, A. L., and R. S. Zucker (1985. Biophys. J. 48: 1003-1017) has been employed as the basis of a refined version of the "Ca theory" for neurotransmitter release. As such, it has been studied here as to its ability to predict the time course of release under various conditions. In particular, conditions were chosen in which the temporal variations in intracellular Ca2+ concentration, the sole factor controlling the release according to the Ca theory, were modified and tested experimentally. The predictions of this model were compared with the experimental results. It is shown that the 3D diffusion model, similarly to earlier simpler versions of the Ca theory, predicts that the time course of release is highly sensitive to both the level of depolarization and the level of the resting concentration of intracellular Ca2+ Moreover, the 3D diffusion model predicts that the time course of release is insensitive to changes in temperature. In contrast, the experimental results show that the time course of release is invariant to the level of depolarization and to the resting level in intracellular Ca2+, but highly sensitive to variations in temperature. PMID- 2566338 TI - Relative bioavailability of almitrine bismesylate in humans. AB - Bioavailability and bioequivalency studies of almitrine bismesylate from U.S. manufactured film coated, waxed, 50 mg tablets were compared in 34 normal healthy volunteers to 50 mg European film coated, waxed and unwaxed, tablets and a 0.5 per cent (w/v) oral reference solution of almitrine bismesylate in d,l malic acid. The U.S. manufactured formulations were 85.88 and 87.85 per cent of the calculated mean area under the individual concentration-time curve for almitrine bismesylate reference solution compared to 88.40 and 88.86 per cent for the waxed and unwaxed film coated European tablets, respectively. The mean peak plasma concentrations for the U.S. formulations were 176.3 ng ml-1 and 180.1 ng ml-1 compared to 196.3 and 200.1 ng ml-1 for the waxed and unwaxed European formulations, respectively. Mean time to peak plasma concentrations for the two U.S. formulations and the waxed and unwaxed European formulations were 3.22, 3.33, 3.06, and 3.26 h, respectively. In addition, the oral reference solution yielded a mean peak plasma concentration of 222.8 ng ml-1 and a mean time to peak plasma concentration of 2.68 h. Analysis of variance and multiple range comparisons (p less than 0.05) indicated that the tablet formulations were bioequivalent. The results of this study show that the U.S. formulated almitrine bismesylate tablets exceed 85 per cent relative bioavailability with respect to the oral reference solution and are bioequivalent compared to the marketed standard European tablet formulations. PMID- 2566339 TI - One year administration of almitrine bismesylate (Vectarion) to chronic obstructive pulmonary disease patients: pharmacokinetic analysis. AB - A double blind study utilizing orally administered almitrine bismesylate was conducted involving 36 stable chronic obstructive pulmonary disease (COPD) patients with hypoxia and with and without hypercapnia. The patients received 50 mg tablets twice daily for 360 days. Blood samples were taken both at predose and 3 hours postdose at different periods throughout 1 year dosage regimen and plasma levels were analyzed by a GLC method using a nitrogen-phosphorous detector. Plasma almitrine concentrations indicate large variability at each time sample. Results suggest an increasing trend in the almitrine plasma levels as a function of time. Plasma almitrine levels increased significantly (p less than 0.01) between test day 14 and test day 360 (243 +/- 213 per cent and 199 +/- 170 per cent for predose and 3h postdose samples, respectively) indicating that steady state is not achieved by day 14. Almitrine plasma levels appear to stabilize between test day 90 and test day 180. The effective multiple dose half-life for almitrine bismesylate in plasma is estimated to be 32 days. About half of the patients exhibited steady state peak plasma almitrine levels above 500 ng ml-1. In addition, 19 per cent of the patients achieved maximum apparent steady state almitrine levels greater than 700 ng ml-1. Mean accumulation was estimated to be 4.21 +/- 1.98 at one year. PMID- 2566340 TI - Effect of the beta-adrenoceptor agonist BRL 26830 on fatty acid synthesis and on the activities of pyruvate dehydrogenase and acetyl-CoA carboxylase in adipose tissues of the rat. AB - BRL 26830 is a thermogenic beta-adrenoceptor agonist which stimulates lipolysis and fatty acid oxidation in vivo. It also stimulates insulin secretion, and hence promotes glucose utilisation in vivo. The effect of this agent on white and brown adipose tissue of the rat was investigated. BRL 26830 increased the rate of fatty acid synthesis in vivo in white adipose tissue by 135% but reduced the rate of fatty acid synthesis in vivo in brown adipose tissue by 78%. The increase was abolished in white adipose tissue of streptozotocin-diabetic rats, indicating that the effect involved a rise in circulating insulin levels. The reduction in fatty acid synthesis in brown adipose tissues was associated with a reduction in the activity of acetyl-CoA carboxylase in the tissue consistent with a direct beta-adrenoceptor-mediated effect. BRL 26830 also increased the proportion of pyruvate dehydrogenase in its active form in vivo in brown adipose tissue and this increase was abolished in streptozotocin-diabetic rats. These findings illustrate different sensitivities of white and brown adipose tissues to combined beta-adrenergic and insulin stimulation. PMID- 2566341 TI - Restriction fragment analysis of the HLA class II region: application to tissue transplantation. AB - Southern blotting and hybridization with locus specific probes is a reliable method for the determination of HLA Class II specificities and in conjunction with several other techniques is providing valuable information on the nature of Class II polymorphisms in man. In terms of the identification of Class II specificities in transplantation however, the advanced nature of DR serology means that DR beta restriction fragment analysis is probably limited only to those situations where serological methods have been unsuccessful. Whether DQ and DP incompatibilities can evoke rejection responses in vivo remains to be fully ascertained although our preliminary data suggest that DQ mismatches probably have little effect upon graft outcome. Restriction fragment analysis is particularly attractive in that it enables reliable retrospective studies to be performed in conjunction with prolonged clinical follow up, and the technique will clearly continue to contribute significantly to our understanding of the importance of these molecules in transplantation. PMID- 2566342 TI - Transglutaminase-catalyzed incorporation of [2,5-3H]histamine into a Mr 84000 particulate protein in pancreatic islets. AB - Rat pancreatic islet homogenates catalyze the incorporation of [2,5-3H]histamine into endogenous proteins recovered in both the stacking gel and a Mr 84000 protein separated by polyacrylamide electrophoresis. The labelling of these proteins represents a Ca2+-dependent process inhibited by glycine methylester, but not sarcosine methylester, and enhanced after preincubation of the islets at a high concentration of D-glucose. Although transglutaminase activity is found in both soluble and particulate subcellular fractions, the endogenous transglutaminase substrates were located mainly in particulate, possibly membrane associated, material. PMID- 2566343 TI - Buprenorphine and temazepam abuse by drug takers in Glasgow--an increase. PMID- 2566344 TI - Rat bites and diabetic foot in the West Indies. PMID- 2566346 TI - World AIDS summit in London. PMID- 2566345 TI - Risks of dependence on benzodiazepine drugs. PMID- 2566347 TI - Fatigue states. PMID- 2566348 TI - Alpha 2-adrenoceptor agonists enhance responses to certain other vasoconstrictor agonists in the rat tail artery. AB - 1. The effects of the alpha2-adrenoceptor agonists clonidine, rilmenidine, TL99 and UK14304 on the vasoconstrictor response to sympathetic nerve stimulation and on the concentration-response curves to noradrenaline and phenylephrine were compared in two isolated, perfused vascular tissues: the rat tail artery (which has both postjunctional alpha 1- and alpha 2-adrenoceptors), and the rabbit ear artery (in which only alpha 1-adrenoceptors are present postjunctionally). 2. In the rabbit ear artery, the first observable effect of alpha 2-adrenoceptor agonists was inhibition of vasoconstrictor responses to sympathetic nerve stimulation. This occurred with concentrations of the alpha 2-adrenoceptor agonists which were far below those producing vasoconstriction. Responses to noradrenaline were not affected. 3. In contrast, in the rat isolated perfused tail artery, alpha 2-adrenoceptor agonists, in concentrations that produced no other observable effects, enhanced the vasoconstrictor responses to sympathetic nerve stimulation and to noradrenaline. Much higher concentrations of alpha 2 adrenoceptor agonists produced vasoconstriction in most preparations and only then reduced the response to sympathetic nerve stimulation. The enhancing effect of alpha 2-adrenoceptor agonists was blocked by idazoxan, but not by prazosin. 4. Vasoconstrictor responses in the rat tail artery to the relatively selective alpha 1-adrenoceptor agonist phenylephrine were enhanced by alpha 2-adrenoceptor agonists. The enhancement of the response to phenylephrine was greater than that to the mixed alpha 1- and alpha 2-adrenoceptor agonist noradrenaline. 5. Vasoconstrictor responses in the rat tail artery to vasopressin, ATP and KCl, like those to alpha 1-adrenoceptor agonists, were enhanced by alpha 2 adrenoceptor agonists.2+owever, vasoconstrictor responses to PMID- 2566349 TI - Evidence for co-transmitter role of neuropeptide Y in the pig spleen. AB - 1. The possible involvement of neuropeptide Y (NPY) in relation to noradrenaline (NA) and adenosine triphosphate (ATP) mechanisms in the sympathetic nervous control of the vascular tone and capsule contraction in the blood perfused pig spleen was investigated in vivo. 2. Local injections or infusions of NA, NPY and alpha-, beta-methylene ATP (mATP) caused vasoconstriction (perfusion pressure increase) and capsule contraction (increased venous blood flow). ATP only evoked vasodilatation. NPY was about 50 fold more potent than NA as a vasoconstrictor, and the NPY response was more long-lasting. Reserpine treatment did not change the effects of NPY. 3. Electrical stimulation of the splenic nerves in control animals caused a frequency-dependent, guanethidine-sensitive output of both NPY like immunoreactivity (-LI) and NA, suggesting co-release. The output of NPY-LI relative to NA was enhanced at high frequency stimulation. Furthermore, alpha adrenoceptor blockade by phentolamine enhanced both the output of NPY-LI and NA while inhibition of the neuronal uptake of NA with desipramine reduced the low frequency stimulation-evoked overflow of NPY-LI. Preganglionic denervation did not change the output of NPY-LI or NA. 4. Reserpine treatment reduced both the splenic content of NA and NPY-LI. Preganglionic denervation inhibited the reserpine-induced depletion of the NPY content but not of NA in terminal areas. The stimulation-evoked NPY overflow was markedly enhanced, especially at low frequency stimulation after reserpine, and the plasma levels of NPY-LI in the venous effluent were then in the nmolar range (i.e. where exogenous NPY induced vasoconstriction). The perfusion-pressure increase upon stimulation in reserpine treated, preganglionically-denervated animals was highly correlated (r = 0.91) to the NPY overflow. The functional 0.5 Hz responses were reduced after reserpine, while at higher frequencies the functional effects were of similar magnitude to controls but longer-lasting. 5. Tyramine induced a release of NA but not of NPY LI. Furthermore, the increase in perfusion pressure induced by tyramine was absent after reserpine. 6. After tachyphylaxis to the vasoconstrictor effects of mATP, the nerve stimulation-evoked, functional response as well as the NA and NPY LI overflow were unchanged. After reserpine treatment, both the perfusion pressure increase and NPY-LI overflow to nerve stimulation were reduced after mATP tachyphylaxis. 7. In conclusion, release of NPY rather than ATP may explain the long-lasting, non-adrenergic, splenic functional responses in reserpinized animals upon sympathetic stimulation. However, NA is most likely the main splenic transmitter when low-frequency stimulation is used under control conditions. PMID- 2566350 TI - The effect of medetomidine, an alpha 2-adrenoceptor agonist, on plasma atrial natriuretic peptide levels, haemodynamics and renal excretory function in spontaneously hypertensive and Wistar-Kyoto rats. AB - 1. The effects of the selective alpha 2-adrenoceptor agonist, medetomidine, were assessed on plasma levels of immunoreactive atrial natriuretic peptide (IR-ANP), haemodynamics and on urine water and solute excretion in conscious, chronically cannulated, 7 month-old spontaneously hypertensive (SHR) and age-matched Wistar Kyoto (WKY) rats, in order to examine the role of alpha 2-adrenoceptors in the control of ANP secretion. 2. A 60 min i.v. infusion of medetomidine (0.2 or 0.6 microgram kg-1 min-1) decreased heart rate dose-dependently in both strains. Medetomidine infusion (0.6 microgram kg-1 min-1) resulted in an increase in mean arterial pressure in WKY, whereas both doses decreased blood pressure in SHR. There was a slight increase in the right atrial pressure in both strains (WKY: +1.18 +/- 0.26 mmHg; SHR: +1.64 +/- 0.64 mmHg, NS) in response to infusion of 0.6 microgram kg-1 min-1 of medetomidine. 3. No differences were found in resting plasma IR-ANP levels between WKY (114 +/- 8 pg ml-1, n = 19) and SHR (117 +/- 10 pg ml-1, n = 21). Infusion of equibradycardic doses of medetomidine increased dose-dependently plasma IR-ANP levels in WKY, but did not affect the plasma IR ANP concentration in SHR rats. 4. Despite the different effect of medetomidine on ANP release in WKY and SHR rats, i.v. administration of medetomidine affected renal excretory functions similarly in both strains; urine flow and sodium excretion increased and urine osmolality decreased significantly, while there was no consistent change in urinary potassium excretion. Urine osmolality decreased to hypo-osmotic levels during the infusion of 0.6 yg kg-1 min1 of medetomidine, suggesting a possible interaction between alpha 2-adrenoceptor stimulation and the vasopressin system. 5. These results show that the alpha 2-adrenoceptor agonist medetomidine increased plasma levels of ANP in WKY rats, probably through an increase in mean arterial and right atrial pressures, whereas the SHR had attenuated ANP release to alpha 2-adrenoceptor stimulation. Our findings, that medetomidine caused marked natriuretic and diuretic effects in both strains and that these effects on the excretory functions of the kidneys were not related to changes in plasma levels of IR-ANP, demonstrate that changes in plasma ANP levels alone do not account for the diuretic and natriuretic effect of alpha 2-agonists. PMID- 2566351 TI - Effect of 5-hydroxytryptamine on [3H]-acetylcholine release from guinea-pig striatal slices. AB - 1. The effect of 5-hydroxytryptamine (5-HT) on spontaneous and electrically evoked tritium efflux was studied in guinea-pig caudate nucleus slices preloaded with [3H]-choline. 2. 5-HT, 10-300 mumol l-1, temporarily increased the spontaneous tritium efflux (as well as the endogenous acetylcholine (ACh) release) and, after 15 min perfusion, inhibited it. The facilitatory effect of 5 HT on spontaneous efflux was increased while the inhibitory effect did not occur in slices taken from dopamine-depleted guinea-pigs. 3. The increase in spontaneous tritium efflux by 5-HT was blocked by methiothepin, methysergide (pA2 8.7) and by the selective 5-HT2 antagonist, ritanserin (pA2 6.7). 4. The inhibition of spontaneous tritium efflux by 5-HT was prevented by methysergide and methiothepin but not by ritanserin and (-)-propranolol. 5. 5-HT, 100 mumol l 1, inhibited the electrically-evoked tritium efflux and this effect was unchanged in dopamine-depleted slices. 6. The inhibition of electrically-evoked tritium efflux by 5-HT was blocked by methiothepin and methysergide but not by (-) propranolol or ritanserin. 7. These results suggest that 5-HT may exert a rapid and transient (excitatory) and a more prolonged (inhibitory) control over striatal cholinergic neurones. PMID- 2566352 TI - Characterization of pigment aggregating alpha 2-adrenoceptors of fish melanophores by use of different agonists after partial irreversible receptor inactivation. AB - 1. The affinity for, and the intrinsic efficacy on, postsynaptic melanosome aggregating alpha 2-adrenoceptors of fish melanophores was studied for B-HT 920, clonidine, medetomidine, noradrenaline, phenylephrine and UK-14,304. Investigations were carried out by evaluating the effects of progressive, irreversible inactivation of the alpha 2-adrenoceptors by benextramine. 2. The double reciprocal plots of equieffective concentrations of B-HT 920, clonidine, noradrenaline and phenylephrine were linear, which indicated that these compounds exerted their effects, mainly, through interaction with one receptor site. 3. The affinity for the alpha 2-adrenoceptor selective agonist B-HT 920, was found to be about 1000 times higher than the affinity for the alpha 1-adrenoceptor selective agonist phenylephrine. 4. The corresponding plot of equieffective concentrations of medetomidine was not linear, which may indicate that this imidazole compound exerted its effect through more than one receptor site. However, when phenoxybenzamine was used in place of the more selective benextramine, a linear relationship was obtained. PMID- 2566353 TI - Lipomobilizing effects of procaterol and yohimbine in the conscious dog: comparison of endocrinological, metabolic and cardiovascular effects. AB - 1. Lipid mobilization during a hypocaloric diet may be enhanced by a pharmacological approach using beta 2-adrenoceptor agonists or alpha 2 adrenoceptor antagonists. Studies were undertaken in the dog, an animal model presenting fat cell antilipolytic alpha 2- and lipolytic beta-adrenoceptors, in order, firstly, to demonstrate the presence of beta 2 subtype adrenoceptors on adipocytes and, secondly, to compare the effects of procaterol (beta 2 adrenoceptor agonist) and of yohimbine (alpha 2-adrenoceptor antagonist) on metabolic, endocrinological and cardiovascular parameters. 2. Procaterol strongly stimulates lipolysis in dog adipocytes in vitro. The utilisation of selective beta 1- and beta 2-adrenoceptor antagonists (bisoprolol and ICI 118,551) in both lipolysis and binding studies (displacement of [3H]-dihydroalprenolol binding) demonstrated the presence of the two beta-adrenoceptor subtypes in dog fat cells. 3. Infusion of either yohimbine or procaterol (10 and 0.4 nmol min-1 kg-1, respectively for 30 min), provoked an equivalent increase in plasma non esterified fatty acids (+100%). Procaterol, but not yohimbine, induced hyperglycaemia (+120%). Plasma insulin was weakly enhanced by yohimbine (+120%) as compared to the increase given by procaterol (+500%). 4. Both drugs stimulated sympathetic nervous system activity, as indicated by the increased plasma noradrenaline concentration, but only yohimbine increased the plasma adrenaline level. 5. Cardiovascular measurements indicated that procaterol strongly enhances heart rate and transiently decreases mean blood pressure. Yohimbine exhibits a weaker effect on heart rate and slightly increases mean blood pressure. 6. The present work clearly indicates that lipid mobilization is enhanced during fasting in the dog by selective beta 2-adrenoceptor stimulation or by alpha 2 adrenoceptor blockade. This enhanced lipolytic effect may result either from a direct action of the drugs on the adrenoceptors of fat cells or from an activation of the sympathetic nervous system. Procaterol suffers major limitations since it strongly increases heart rate, immunoreactive insulin and glycaemia. On the other hand, yohimbine induces only minor modifications both in cardiovascular and endocrinological parameters. PMID- 2566355 TI - Changing pattern of admissions and operations for duodenal ulcer. AB - The admission rates for duodenal ulcer (DU) and the effect of H2-receptor antagonists (H2RA), introduced in the Trent Region of the UK in 1977, were examined. The admission rates are expressed per 10(6) of resident population. The use of H2RA has risen 3.7-fold (from 1978 to 1983), yet overall admission rates for perforation have changed little: 99 in 1972-76 (pre-H2RA period) compared with 103 in 1977-84 (H2RA period). Admission rates for haemorrhage have risen by 8 per cent, from 130 to 140 (P less than 0.01). However, the overall rates conceal large increases (P less than 0.01) in the admission rates for those aged greater than or equal to 65 years, of 33 per cent (from 264 to 352) for perforation and of 28 per cent (from 381 to 489) for haemorrhage. Emergency admissions for uncomplicated DU were unchanged: 88 in 1972-76 and 89 in 1977-84. However, the proportions operated on fell by 58 per cent (P less than 0.01), from 30 per cent of admissions in the pre-H2RA period compared with only 12 per cent in the H2RA period. Waiting-list admissions for uncomplicated DU fell by 43 per cent, from 187 to 106 (P less than 0.01), and the proportions operated on fell from 162 to 76; the combined effect resulted in a reduction of 53 per cent in the operation rates (P less than 0.01). In Rotherham, the use of H2RA has risen 6.2 fold (from 1978 to 1983) and they were generally used intermittently in 1976-78 and later for maintenance therapy and high-dose treatment. Yet admissions for perforation and for haemorrhage were unchanged. Emergency admissions for uncomplicated DU rose by 40 per cent, from 130 in 1972-75 to 182 in 1976-84, but the proportions operated on fell markedly, from 20 to 6 per cent (P less than 0.01); waiting-list admissions fell in 1976-78 by 29 per cent and in 1979-84 by 73 per cent. The proportions operated on in the three periods fell from 74 to 53 per cent and 25 per cent respectively and these two factors led to decreases in elective surgery of 50 per cent in 1976-78 and 91 per cent in 1979-84 (P less than 0.01).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2566354 TI - 6-Cyano-7-nitroquinoxaline-2,3-dione as an excitatory amino acid antagonist in area CA1 of rat hippocampus. AB - 1. A quantitative pharmacological investigation of the excitatory amino acid antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) has been made in area CA1 of rat hippocampal slices bathed in 1 mM Mg2+ containing medium. 2. At a concentration of 10 microM, CNQX reversibly antagonized responses to alpha-amino 3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), quisqualate and kainate; it produced a parallel shift in their log dose-response curves. Responses to N methyl-D-aspartate (NMDA) were not antagonized by 10 microM CNQX (dose-ratio: 1.04 +/- 0.06, n = 3). 3. Schild plots (constructed over the range 1-100 microM) yielded the following estimated pA2 values, AMPA 5.8, quisqualate 5.9, and kainate 5.9. NMDA was antagonized by 100 microM CNQX, giving an apparent log K of 4.44 +/- 0.06. 4. The slopes (+/- s.e. mean) of the Schild plots were for AMPA 0.84 +/- 0.06, quisqualate 0.79 +/- 0.04 and kainate 0.68 +/- 0.07. These were all significantly less than unity. 5. Synaptic responses elicited by low frequency activation of the Schaffer collateral-commissural pathway were blocked completely by CNQX (10 microM) providing that a low stimulus intensity was used. With high intensity stimulation a small component remained that was blocked by the selective NMDA antagonist D-2-amino-5-phosphonovalerate (APV). 6. These results suggest that CNQX does not differentially affect the responses of CA1 neurones to AMPA, quisqualate and kainate. It does, however, depress responses to these agonists to a greater degree than it does responses to NMDA and it is a highly effective synaptic antagonist. PMID- 2566356 TI - Are there bulbospinal catecholaminergic neurones in the guinea pig equivalent to the C1 cell group in the rat and rabbit? AB - The C1 cell group in the rat is characterized by neurones which contain both adrenaline and phenylethanolamine-N-methyltransferase, and usually also neuropeptide Y (NPY). The former two substances are lacking in Guinea pig brainstem and spinal cord. We have examined the distribution of NPY- and tyrosine hydroxylase-immunoreactivity in the ventrolateral medulla and thoracolumbar intermediate zone of Guinea pig, as well as the distribution of catecholamine containing neurone somata and spinal terminals visualized after formaldehyde glutaraldehyde fixation. The results are compared with comparable immunohistochemical data obtained from rats and rabbits. Catecholaminergic neurones in the Guinea pig with locations and terminations that correspond to those of the C1 cell group in rat and its analogue in the rabbit appear to consist of two subgroups, with only the more caudal group containing NPY. The more rostral group requires pretreatment with monoamine oxidase inhibitor to permit visualization of catecholamine fluorescence, a property previously though to be characteristic of adrenergic neurones. This observation raises the possibility that the catecholaminergic cell group in the C1 region of rabbits may not contain adrenaline either. PMID- 2566357 TI - Accelerated norepinephrine turnover in peripheral tissues after ventromedial hypothalamic stimulation in rats. AB - To obtain evidence for a functional connection between the ventromedial hypothalamic nucleus (VMH) and the sympathetic nervous system, effects of electrical stimulation of the VMH, the lateral hypothalamic area (LH) and the paraventricular hypothalamic nucleus (PVN) on norepinephrine (NE) turnover in the heart, liver, pancreas, spleen, submandibular gland and the interscapular brown adipose tissue were examined in anesthetized rats. Stimulation of the VMH elicited a 3-8-fold increase in the rate of NE turnover in all organs examined, whereas stimulation of the LH or the PVN had no appreciable effects. The effect of VMH stimulation was abolished after sympathetic ganglionic blockade with hexamethonium. Epinephrine turnover in the adrenal gland was accelerated by stimulation of not only the VMH but also the LH. It was concluded that the VMH is intimately associated with sympathetic facilitation in peripheral tissues. PMID- 2566358 TI - Organizational role of testosterone on the biochemical and morphological development of the hypogastric ganglion. AB - Previous reports have demonstrated that after postnatal day 10 testosterone influences hypogastric ganglion (HG) development by 'activating' morphological and biochemical indices. We now report an 'organizational' influence on the developing HG during the first 10 postnatal days. To investigate the organizational effects of testosterone, male rats were castrated within 12 h of birth. Testosterone replacement therapy initiated following castration maintained the normal number of neurons in the HG. Conversely, delayed replacement therapy starting at day 10 or vehicle treatment only, resulted in a significant decrease in neuron number. Castration also produced a significant decrease in somal and nuclear cross-sectional areas. Testosterone replacement, whether initiated immediately or if delayed until day 10, restored somal and nuclear cross sectional areas to normal. Tyrosine hydroxylase (TH) and choline acetyltransferase (ChAT) activities were sensitive to both testosterone dosage and the time of administration. Testosterone decanoate administered subsequent to castration was not able to completely reverse the enzyme activity deficits, while delayed replacement therapy was even less effective in restoring enzyme activities. In contrast, higher doses of testosterone completely reversed enzyme activity deficits, and in fact produced a significant increase in TH activity. Again, delayed testosterone replacement did not fully restore deficits in enzyme activity. In summary, the hormonal environment during the first 10 days of life is critical for the organization of HG cell number; in contrast, nuclear and cell size appear to be dependent on testosterone for activation. TH and ChAT activities also appear to be organized during this dose- and time-dependent developmental period. PMID- 2566360 TI - Stimulation-dependent release of adenosine triphosphate from hippocampal slices. AB - Schaffer collaterals of rat and mouse hippocampal slices were stimulated with bursts of pulses (300 Hz for 50 ms, 2-s intervals) for 30-s which caused a stable increase in the size of the population spike known as long-term potentiation. The release of adenosine triphosphate (ATP) was measured with a luciferase-luciferine system and the light emitted was recorded with a photomultiplier placed beneath a modified slice chamber. ATP release was observed shortly after the start of stimulation and was quantified by comparison with the response of standard solutions of ATP. No ATP release was observed in a Ca2+ free solution or after low frequency stimulation (1 Hz). Glutamate (2 mM), applied without electrical stimulation, did not evoke ATP release. Also, the glutamate receptor blocker, kynurenic acid (10 mM), did not block ATP release. It is concluded that ATP is released from electrically stimulated hippocampal slices from presynaptic nerve terminals in a calcium-dependent fashion and may play a role in the modulation of synaptic efficiency. PMID- 2566359 TI - Hypogastric ganglion perinatal development: evidence for androgen specificity via androgen receptors. AB - The hypogastric ganglion (HG) has previously been shown to be sensitive to both the organizational and activational influences of testosterone. The current investigations examined whether testosterone exerts similar effects prenatally, whether these events are specifically controlled by androgen, and whether androgens might directly masculinize the HG. Prenatal treatment with an anti androgen, flutamide, resulted in significant decreases in the adult levels of tyrosine hydroxylase (TH) activity, an index of postsynaptic noradrenergic ontogeny, and choline acetyltransferase (ChAT) activity, a marker for presynaptic terminal formation. In addition, testosterone propionate and dihydrotestosterone benzoate reversed the effects of neonatal castration on the development of TH and ChAT activities. In contrast estradiol benzoate was unable to restore enzyme activities. To determine whether the above observations might be produced by direct effects on the HG, androgen cytosol receptor characteristics were studied. Competition and saturation analyses demonstrate that the affinity and specificity of the androgen cytosol receptor in the HG are similar to that displayed in the pituitary, which has previously been shown to contain androgen receptors. These results suggest that the adult levels of TH and ChAT activities are organized during prenatal and early postnatal development. In addition, the organization of the HG appears to be androgen specific. The presence of cytosol androgen receptors suggests that the organizational effects of androgens are possibly induced by a direct mechanism. PMID- 2566361 TI - Characterization of stimulation-produced analgesia from the nucleus tractus solitarius in the rat. AB - Electrical stimulation of the commissural region of the nucleus tractus solitarius (NTS) inhibits the tail-flick reflex evoked by noxious heat. This antinociception can be measured in the awake or pentobarbital anesthetized rat at current intensities that do not induce overt behavioral side effects. Glutamate microinjections into the NTS, but not immediately surrounding the NTS, also inhibit the tail-flick reflex, demonstrating that activation of NTS cell bodies, and not fibers of passage, mediates antinociception from this region. In contrast, morphine microinjections into the NTS have no effect on the tail-flick reflex in anesthetized rats. These findings provide further evidence that the NTS is involved in the modulation of nociception. PMID- 2566362 TI - Nucleus raphe obscurus (nRO) influences vagal control of gastric motility in rats. AB - Because the nucleus raphe obscurus (nRO) maintains a direct connection with the dorsal vagal complex in the medulla, this nucleus has the potential to influence vagal control of gastric function. Both electrical- and glutamate-induced activation of the nRO were found to enhance gastric motility and tone in the rat. The gastric responses to nRO stimulation were abolished by peripheral muscarinic blockade. PMID- 2566363 TI - Auditory nerve neurotransmitter acts on a kainate receptor: evidence from intracellular recordings in brain slices from mice. AB - Intracellular recordings from neurons in brain slice preparations of the mouse ventral cochlear nucleus (VCN) were used to examine the actions of excitatory amino acid agonists and antagonists. Synaptic responses to electrical stimulation of the auditory nerve root were partially blocked by kynurenic acid, an antagonist that is specific for glutamate receptors. The antagonists specific for N-methyl-D-aspartate (NMDA), DL-2-amino-5-phosphonovalerate (APV) and Mg2+, did not affect the response, arguing against a role for NMDA receptors at the VIIIth nerve synapse. To test postsynaptic sensitivity to excitatory amino acid agonists, responses to bath applications were measured in VCN neurons while synaptic transmission was blocked by the removal of Ca2+ from the bath or by the addition of tetrodotoxin. Neurons in the VCN were 500-1000 times more sensitive to kainate than to glutamate or aspartate. In the absence of Mg2+, they were also sensitive to NMDA. The responses to kainate and glutamate were increased by the removal of calcium from the bath. These results imply that VCN neurons have both kainate and NMDA receptors and that synaptic transmission between auditory nerve fibers and neurons in the cochlear nuclear complex could be mediated by a substance related to kainate. PMID- 2566365 TI - Electrophysiological and biochemical characterization of a continuous human astrocytoma cell line with many properties of well-differentiated astrocytes. AB - Astrocytes comprise about 25% of the cellular volume of the brain, and their main function is to maintain homeostasis of the neuronal environment. These cells are commonly identified on the basis of their membrane electrical properties and the presence of specific proteins. We have characterized the human astrocytoma cell line designated UC-11MG and have shown these cells have many of the traits of differentiated astrocytes. Many of the UC-11MG cells have a large resting membrane potential, averaging -74 mV. The slope of the Em vs log [K]o cuve was 58.5 mV per decade [K]o. The cells were inexcitable when exposed to brief depolarizing current pulses. The astrocytoma traits are virtually identical to those previously reported for normal astrocytes. The astrocytoma cells also express glutamine synthetase activity which is considered specific to astrocytes among brain cells. Previous work had also demonstrated the presence of other astrocyte markers glial fibrillary acidic protein and S-100 protein in the UC 11MG cells. The steady-state ion transport properties of Na+, Cl-, and K+ were also characterized in these cells, and the rates of efflux were found to be similar to those in other astrocytes, with the major difference being the presence of a second kinetic compartment in the UC-11MG cells. From this work, we conclude that the UC-11MG cell line displays prominent features associated with differentiated astrocytes, and may provide an excellent model system for the study of human astrocytes. PMID- 2566364 TI - Effects of potassium channel blockade on endogenous glutamate release from cerebellar slices. AB - The effects of potassium channel blockade on the spontaneous release of endogenous glutamate from rat cerebellar slices was studied. Tetrapentylammonium (TPeA), 4-aminopyridine and quinine all increased the spontaneous release of glutamate. The effect of TPeA and 4-AP was potentiated in the absence of Ca2+ from the perfusing fluid. In normal artificial cerebrospinal fluid (ACSF) the Ca2+ channel antagonist, verapamil, mimicked the effects of TPeA seen in Ca2+ free ACSF. The increased release of glutamate produced by TPeA under Ca2+-free conditions was inhibited by the sodium channel blocker, tetrodotoxin, and by Ruthenium red, which inhibits mobilization of mitochondrial Ca2+. The results suggest that external Ca2+ is not required in the TPeA-induced release of glutamate. It is proposed that the prolongation of depolarization seen with potassium channel blocking drugs enables sufficient sodium to enter the neurone and release Ca2+ from intraneuronal stores in order to facilitate transmitter release. PMID- 2566366 TI - [Risk factors and prevention of tardive dyskinesias]. AB - Tardive dyskinesia is a frequent complication of a neuroleptic treatment. The physiopathology is unclear and the curative treatments inadequate. This explains the necessity for preventive measures at three levels: the prescription of neuroleptics, the evaluation of risk factors and early diagnosis. An analysis of epidemiological studies allows the evaluation of risk factors published in literature. PMID- 2566367 TI - Pharmacotherapy of schizophrenia. AB - Numerous advances in our understanding of schizophrenia and its pharmacotherapy have occurred over the last forty years, reflected in basic research endeavours exploring the pathophysiology of schizophrenia, as well as efforts to establish new medications with a more selective psychotropic action and fewer side effects. Ongoing clinical research has, at the same time, provided valuable information regarding the applicability of the various agents, appropriate dosing, symptom response patterns and side effect profiles. This growth in knowledge also has served to underscore the limitations and drawbacks of currently available drugs, leading to a more cautious approach in their use. Newer technologies, such as positron emission tomography, hold considerable promise in our efforts to further clarify the precise mechanisms mediating schizophrenia, and to establish treatment alternatives. PMID- 2566368 TI - The immunophenotype of Reed-Sternberg cells. A study of 50 cases of Hodgkin's disease using fixed frozen tissues. AB - The authors analyzed 50 cases of Hodgkin's disease (HD) with a panel of antibodies which detect B-cell and T-cell specific markers and activation antigens using a sensitive immunocytochemical technique and paraformaldehyde lysine-periodate (PLP) fixed-frozen tissues. In 60% of cases either T-cell or B cell specific antigens were detected on Reed-Sternberg (RS) cells. Most T-cell cases were of nodular sclerosing (NS) and mixed cellularity (MC) type (65% and 30%, respectively) and most B-cell cases were either of NS or lymphocyte predominant (LP) type (55% and 45%, respectively). Leukocyte common antigen (LCA) was usually negative on RS cells in NS, but was present in approximately 50% of the cases of MC and LP types. Almost all cases were positive for the CD30 antigen (Ki-1). Most cases were also positive for CD15 (LeuM1) with the exception of the LP type. Activation antigens (Ia, CD25, T9) were expressed in a high proportion of cases regardless of subtype. The results suggest that most cases of HD are histogenetically derived from activated T-cells or B-cells. PMID- 2566369 TI - Controversies in cancer pain. Medical perspectives. AB - The treatment of pain in the patient with cancer has focused attention on a series of controversial issues involving medical, social, and moral factors. The medical factors include a lack of knowledge on the part of health care professionals regarding the rational use of opioid drugs. This is coupled with real limitations in the general understanding of the mechanisms of pain and its treatment using pharmacologic, anesthetic, and neurosurgical approaches. Several pharmacologic controversies, including the choice of drug, route and method of administration, and tolerance development and risk of substance abuse, have emerged with the use of opioids on a chronic basis in the cancer population. The social and moral implications involve the issue of who will pay for high technology pain management approaches for patients either at home or in hospice care and the ethical considerations in managing pain with opioid drugs. Carefully designed studies to assess these factors, coupled with broad educational programs, will improve the care of cancer patients in pain and expand our understanding of these important issues. PMID- 2566370 TI - Continuity of care for the cancer patient with chronic pain. AB - The care of patients with cancer and chronic pain, often complicated and demanding, strains the resources of a single physician. Continuity of care programs, such as that developed by the Pain Service of Memorial Sloan-Kettering Cancer Center (New York), with good communication and liaison work between hospital and community, add a much needed dimension to the pain management of these patients in the home. Although continuity of care programs resemble hospice programs in philosophy, there are major differences in admission criteria: the program is not restricted to dying individuals; patients may live alone; they may receive active therapy with a focus on cure or remission; and they continue to receive their care, including pain management, within a standard medical system under the supervision of their primary physician and nurse. PMID- 2566371 TI - Management of cancer pain. Pharmacology and principles of management. AB - The pain of the vast majority of patients with cancer can be controlled with the use of analgesic drugs. Pharmacokinetic and clinical studies have provided the basis for guidelines for the pharmacologic management of cancer pain. The fundamental concept underlying these guidelines is individualization of therapy, which has as its goal the maximization of pain relief and the minimization of adverse drug effects. This article presents these guidelines and discusses the pharmacologic properties and adverse effects of analgesic drugs commonly used in the treatment of cancer pain. Also reviewed are methods for avoiding and treating the adverse effects of analgesics. PMID- 2566372 TI - Anesthetic techniques for the management of cancer pain. AB - This review focuses on available anesthetic techniques for cancer patients, the indications, and appropriate agents for these potent tools in a stepwise approach to cancer pain. Anesthetic procedures are desirable when they will not compromise bodily functions important to the patient, and when tumor-directed therapy and noninvasive or less-invasive, low-risk approaches (primarily pharmacologic tailoring of analgesic drugs) fail to control pain. Nondestructive techniques include the epidural/intrathecal use of opioids via an implanted catheter, and local anesthetic blocks of nerves and sympathetic ganglia. Chronic intrathecal or epidural opioid infusion seems ideal if the life expectancy is 2 to 3 months. Local anesthetic blocks can help prognosticate results of subsequent neurolytic blocks, including undesirable effects. Destructive anesthetic procedures comprise injections of neurolytic agents (most commonly phenol or alcohol), and insertion of freezing probes, into nerves and ganglia. The types of nerve blocks performed, their complications, and success rates, and limitations of commonly used neurolytic agents as well as their proper applications, are described. The importance of proper patient selection and knowledge of the pathophysiology of the pain being treated is stressed, as is the appropriate timing of anesthetic procedures in the course of the disease. PMID- 2566373 TI - Neurosurgery in the treatment of cancer pain. AB - Patients with pain syndromes resulting from recurrent or metastatic cancer should be evaluated carefully to determine the cause of their pain and the need for appropriate antitumor treatment. Currently, opioid analgesics are the mainstay of pain control, but side effects limit their use in some patients. When pharmacologic pain control is inadequate or associated with intolerable side effects, neurosurgery should be considered. Currently the implantation of a pump for the intraspinal infusion of opioid analgesics is the most popular procedure, but its usefulness may be limited by the development of opioid tolerance. The most effective ablative pain control procedure at the current time is cordotomy, which is indicated in patients with unilateral pain. Although the place of neurostimulatory procedures in controlling cancer pain is not well established, they are attractive because of their nondestructive nature and potential usefulness in the treatment of bilateral pain syndromes. Specific antitumor surgical procedures should be considered in patients with certain spinal and plexopathy syndromes, because such intervention offers the prospect of both pain relief and tumor control. In this article, the neurosurgical procedures used in the management of cancer pain are reviewed. PMID- 2566374 TI - Pain management in a drug-oriented society. AB - A drug-oriented society promotes drug treatment of illness but responds with restrictive legislation and mores when faced with serious drug abuse by the populace. Narcotics are currently controlled because of their history of abuse and associated crime, and when their use, for whatever purpose, is suspected of abuse. Inadequate treatment of cancer pain with narcotics may stem, in part, from these events. Its major causes are confusion of legitimate and illegitimate narcotics use by society in general and health care providers in particular, combined with undefined terms (e.g., addicts, habitual users) primarily in state statutes, which should be revised and standardized. These factors intimidate physicians from prescribing rationally. For example, a prescription for an extremely large dose, and ordering an adequate quantity of a drug to have at home for a reasonable period of time, as is often required for the control of severe pain, may be perceived as an invitation for investigation of the physician's legitimacy. The real or imagined fear of an investigation encourages him or her to write prescriptions for multiple narcotics, each at the "acceptable" dosage, rather than for single narcotics in larger doses, which is simpler for the patient and preferable from a medical standpoint. Drug abuse is not generally a problem among cancer patients with pain. Physicians should strive to change social attitudes toward pain control with narcotics by enlisting the support of colleagues and, if necessary, by political activism. PMID- 2566375 TI - Structure and function of pili of pathogenic Neisseria species. PMID- 2566376 TI - Correlation of multidrug resistance with decreased drug accumulation, altered subcellular drug distribution, and increased P-glycoprotein expression in cultured SW-1573 human lung tumor cells. AB - Four multidrug-resistant variants of the human squamous lung cancer cell line SW 1573 with levels of doxorubicin resistance ranging from 10- to 2000-fold were characterized with respect to drug accumulation and efflux, subcellular drug distribution pattern, antioxidant defenses, and P-glycoprotein expression. For all these parameters except the antioxidant defenses a correlation was observed with the level of doxorubicin resistance; with increasing drug resistance cellular drug accumulation capacity (as measured for doxorubicin) progressively decreased, initial drug efflux rates (as measured for daunorubicin) progressively increased, while the subcellular doxorubicin distribution (as measured by fluorescence microscopy) gradually shifted from a "mainly nuclear" to a "mainly cytoplasmic" pattern. Our data suggest that in the present set of cell lines the same mechanism of resistance is operating at all levels of doxorubicin resistance. PMID- 2566377 TI - Correlation between long-term survival in breast cancer patients and amplification of two putative oncogene-coamplification units: hst-1/int-2 and c erbB-2/ear-1. AB - The incidence and association with 10-year survival of amplification in five protooncogenes or transforming genes were retrospectively examined using DNAs extracted from formalin-fixed, paraffin-embedded blocks of tissues obtained from 176 consecutive patients surgically treated for primary breast carcinoma. The incidences of greater than threefold amplification of hst-1, int-2, c-erbB-2, ear 1 (one of c-erbA), and c-myc were 12, 13, 16, 10, and 4.0%, respectively. hst-1 and int-2 were almost always coamplified (21/22), while c-erbB-2 and ear-1 were frequently coamplified (18/28) with almost the same copy number. The hst-1 and int-2 pair and the c-erbB-2 and ear-1 pair, localized on chromosomes 11q13 and 17q21-22, respectively, in normal cells, were inferred to be constituents of different amplification units. Amplification of hst-1 and/or int-2 was detected preferentially in the younger age group, and was correlated with poorer prognosis in cases carrying four or more copies of the genes. Amplification of c-erbB-2 and/or ear-1 was strongly correlated with poor prognosis in all 176 patients, especially those with lymph node metastasis. Amplification of c-myc was also correlated with poor prognosis. Cox's life-table regression analysis showed that amplification of c-erbB-2 had a prognostic value, which was independent of other known prognostic factors such as lymph node status and tumor size. PMID- 2566378 TI - Correlation between reversing of multidrug resistance and inhibiting of [3H]azidopine photolabeling of P-glycoprotein by newly synthesized dihydropyridine analogues in a human cell line. AB - Ten synthetic dihydropyridine analogues were investigated for their ability to reverse drug resistance in a multidrug-resistant human carcinoma cell line, KB Cl. Four dihydropyridine analogues completely reversed the resistance, three lowered the resistance, and three had little effect. The radioactive photoactive dihydropyridine calcium channel blocker, [3H]azidopine, photolabels P glycoprotein in membrane vesicles from KB-Cl cells. This photolabeling was almost completely inhibited by excess dihydropyridine analogues that reversed or lowered drug resistance. In contrast, the labeling was not significantly inhibited by analogues that do not reverse resistance. Among other reversing agents, cepharanthine and reserpine inhibited the [3H]azidopine photolabeling, but thioridazine did not. N-Solanesyl-N,N'-bis(3,4-dimethoxybenzyl)ethylenediamine slightly inhibited the labeling at 100 microM. An anticancer agent, vinblastine, also inhibited the labeling. The correlation between the reversing of the drug resistance and the inhibition of the [3H]azidopine photolabeling of P glycoprotein by dihydropyridine analogues suggests a role for P-glycoprotein in multidrug resistance and also the reversing of the resistance by dihydropyridine analogues. PMID- 2566379 TI - Characterization of monoclonal antibodies recognizing a Mr 180,000 P glycoprotein: differential expression of the Mr 180,000 and Mr 170,000 P glycoproteins in multidrug-resistant human tumor cells. AB - P-glycoprotein is a plasma membrane protein believed to mediate resistance to natural product drugs such as vincristine, Adriamycin, and actinomycin D. To facilitate the study of human P-glycoprotein, monoclonal antibodies (designated HYB-612, HYB-241, and HYB-195) were raised against vincristine-resistant human neuroblastoma (SH-SY5Y/VCR) cells. The antibodies recognize a Mr 180,000 plasma membrane phosphoglycoprotein produced in increased amounts in SH-SY5Y/VCR as well as in vincristine-resistant human neuroepithelioma (MC-IXC/VCR), vinblastine resistant human leukemia (CEM/VLB100), and actinomycin D- or vincristine resistant Chinese hamster (DC-3F/AD X and DC-3F/VCRd-5L) cells, as compared to control cells. Radioimmunoprecipitation of proteins in cells metabolically labeled with [35S]methionine, 32Pi, or [3H]glucosamine and Western transfer procedures were used for these studies. Characterization of the HYB-612 or HYB 241 antigen by destructive degradation produced a pattern of results typical of a conformation-dependent protein epitope. HYB-612 recognizes complexes of the Mr 180,000 antigen with an iodinated photoaffinity analogue of vinblastine or with tritiated azidopine. Furthermore, pretreatment of MC-IXC and MC-IXC/VCR cells with HYB-612 or HYB-241 before measurement of tritium-labeled actinomycin D or vincristine uptake increases the amount of drug accumulation in resistant, but not in sensitive, cells. Of importance is the fact that the Mr 180,000 protein is expressed in cells which also contain a Mr 170,000 P-glycoprotein. The relative amounts of the Mr 180,000 and 170,000 species vary from one drug-resistant cell line to another. Evidence that the Mr 180,000 protein is a P-glycoprotein and that there is a conserved complex pattern of resistance-related surface proteins in multidrug-resistant cells is presented in this report. PMID- 2566380 TI - Differences between the promoting activities of the peroxisome proliferator WY 14,643 and phenobarbital in rat liver. AB - In order to characterize the promoting activity of the peroxisome proliferator [4 chloro-6-(2,3-xylidino)-2-pyrimidinylthio]acetic acid (WY-14,463), male F344 rats which received a single 150-mg/kg dose of diethylnitrosamine (DEN) were fed 0.1% WY-14,643 or 0.05% phenobarbital in the diet for 11, 22, or 54 wk. WY-14,643 promoted the development of ATPase-deficient foci but not GGTase-positive or G6Pase-deficient foci, in contrast to phenobarbital which promoted development of foci detected by all three markers. The mode of promotion of ATPase-deficient foci by WY-14,643 was distinctly different from that of phenobarbital. WY-14,643 primarily increased mean volume of foci at 11 and 22 wk, while phenobarbital primarily increased the numerical density of foci at these time points. At 54 wk, the yield of hepatic neoplasms per liver was higher in rats fed WY-14,643 than in rats fed phenobarbital. To evaluate the possibility that the promotional activity of WY-14,643 was more effective at a later stage in hepatocarcinogenesis, rats receiving a dose of DEN and then phenobarbital in the diet for 11 wk were changed to a diet containing WY-14,643 for an additional 11 or 43 wk. However, WY-14,643 feeding from wk 11 to 22 caused a reduction in volume density of ATPase-deficient foci relative to the volume density of foci at 11 wk. In addition, feeding WY 14,643 from wk 11 to 54 caused similar yields of hepatic neoplasms whether or not phenobarbital was fed for the initial 11 wk. WY-14,643 induced hepatic peroxisome proliferation as indicated by palmitoyl CoA oxidase activity regardless of prior treatment with DEN and/or phenobarbital. The yield of neoplasms in rats not receiving DEN was greater in rats fed WY-14,643 for wk 11 to 54 than in rats fed WY-14,643 for wk 1 to 54. In summary, the peroxisome proliferator WY-14,643 was a more efficient promoter of hepatocarcinogenesis in DEN-initiated rats than phenobarbital. The promotional activity of WY-14,643, when evaluated by stereological analysis and by changing promoters, is distinct from that of phenobarbital, perhaps suggesting different cellular and/or molecular mechanisms of promotion. Understanding the role of promotion by WY-14,643 and other peroxisome proliferators may be important in understanding the mechanism of their hepatocarcinogenicity. PMID- 2566381 TI - Guanfacine treatment of Alzheimer's disease. PMID- 2566382 TI - Hox and HOM: homologous gene clusters in insects and vertebrates. PMID- 2566383 TI - The murine and Drosophila homeobox gene complexes have common features of organization and expression. AB - In situ hybridization analysis of mouse embryos shows the seven members of the Hox-2 complex to be differentially expressed in the central and peripheral nervous system and in mesodermal derivatives (somites and lung). Beginning at the 5' end of the cluster, each successive gene displays a more anterior boundary of expression in the central nervous system. A gene's position in the Hox-2 cluster therefore reflects its relative domain of expression along the anteroposterior axis of the embryo, a feature observed with Drosophila homeotic genes. Sequence comparisons of the Hox-2 cluster with other mouse and Drosophila homeobox genes have defined subgroups of related genes; in the mouse there are four clusters related by duplication and divergence. Alignment shows a clear relationship among genes in the mouse and Drosophila complexes, based on relative position, sequence identity, and domains of expression along the rostral-caudal axis. Our results argue that these complexes arose from a common ancestor, present before the divergence of lineages that gave rise to arthropods and vertebrates. PMID- 2566384 TI - Cooperative binding of steroid hormone receptors contributes to transcriptional synergism at target enhancer elements. AB - We demonstrated previously that two molecules of steroid hormone receptor bound efficiently to a single hormone response element (GRE/PRE) of the tyrosine aminotransferase gene (Tsai et al., 1988). Here, we show that two tandemly linked GRE/PREs conferred progesterone inducibility synergistically to a heterologous TK CAT fusion gene. Binding studies demonstrated that occupation of one GRE/PRE site by a progesterone receptor dimer increased the binding affinity of receptors for the second GRE/PRE site 100-fold. Thus, the observed synergistic induction of TK CAT may result from cooperative binding of receptor dimers to the two GRE/PRE sites. PMID- 2566386 TI - The homeotic gene fork head encodes a nuclear protein and is expressed in the terminal regions of the Drosophila embryo. AB - The region-specific homeotic gene fork head (fkh) promotes terminal as opposed to segmental development in the Drosophila embryo. We have cloned the fkh region by chromosomal walking. P element-mediated germ-line transformation and sequence comparison of wild-type and mutant alleles identify the fkh gene within the cloned region. fkh is expressed in the early embryo in the two terminal domains that are homeotically transformed in fkh mutant embryos. The nuclear localization of the fkh protein suggests that fkh regulates the transcription of other, subordinate, genes. The fkh gene product, however, does not contain a known protein motif, such as the homeodomain or the zinc fingers, nor is it similar in sequence to any other known protein. PMID- 2566385 TI - Development of the Drosophila retina: inductive events studied at single cell resolution. PMID- 2566387 TI - Inhibitory effects of anti-CD2 monoclonal antibodies on interleukin 2 production and interleukin 2 receptor expression in anti-CD3-induced T cell activation. AB - The effects of anti-CD2 monoclonal antibodies (mAb) on anti-CD3-driven interleukin 2 (IL2) production and IL2 receptor (IL2R) expression were investigated. Two anti-CD2 mAb, which had previously been shown to inhibit in vitro anti-CD3-induced T cell proliferation, also inhibited anti-CD3-induced IL2 production. However, it seemed unlikely that this was the crucial mechanism in the inhibition of anti-CD3-driven proliferation, since anti-CD2 mAb also partially inhibited T cell proliferation induced by the anti-CD3 mAb 446 which does not induce detectable IL2 levels. Anti-CD2 mAb also inhibited anti-CD3 induced surface IL2R expression as measured by immunofluorescence staining with an anti-IL2R mAb against the p55 chain. Inhibition of IL2R expression paralleled inhibition of proliferation. This anti-CD2-mediated inhibition involved a block in the generation of normal numbers of IL2R+ cells rather than a direct inhibitory effect on the IL2R+ cells themselves, since IL2R+ cells isolated from anti-CD2-containing cultures responded normally to IL2. Exogenous IL2 and IL4, singly or in combination, could reverse neither the anti-CD2-mediated inhibition of anti-CD3-induced proliferation nor the anti-CD2-mediated inhibition of anti CD3-induced IL2R expression. Taken together, these observations suggest that anti CD2 mAb inhibit anti-CD3-driven proliferation by inhibiting the generation of IL2R+ cells at a maturational stage proximal to their expression of surface IL2R. This inhibition cannot be overcome by exogenous IL2 or IL4, suggesting that the underlying biochemical mechanism involves an IL2- and IL4-independent pathway. PMID- 2566388 TI - Detection of a Thy-1 precursor in human T lymphoma cell lines. AB - In an attempt to gain insight into the biosynthesis and processing of human Thy 1, rabbit antisera (R alpha TP) were raised against a synthetic peptide (TP) of 13 amino acid residues of identical amino acid sequence to that of residues 110 122 of the putative Thy-1 precursor deduced from the cDNA sequencing. Immunoblotting assay showed that the IgG fraction of R alpha TP (R alpha TP-IgG) recognized the synthetic peptide without demonstrable cross-reactivity with isolated human brain Thy-1 and detergent-solubilized membrane proteins of human brain cells. Immunohistochemical studies showed that both R alpha TP-IgG and HB 2S-1 (a monoclonal antibody which reacts with both membrane-bound Thy-1 on viable cells and detergent-solubilized Thy-1) stained cells of two human T lymphoma cell lines (HUT-78 and HUT-102) but they did not stain cells of a human B lymphoma cell line (Raji). In contrast, in cell surface indirect immunofluorescence assay, HB-2S-1 reacted with HUT-78 and HUT-102 cells while R alpha TP-IgG did not. Taken together, these data indicate the existence of a precursor form of human Thy-1 which is processed prior to anchoring to the cell surface. PMID- 2566389 TI - [The functioning of cryptorchid testes in puberty and adolescence]. PMID- 2566390 TI - Spectroscopic analysis of charge transfer complex formation between neuroleptics and iodine. AB - Molecular interactions between iodine and various neuroleptics were investigated by UV/Vis spectroscopy. Iodine was found to form charge transfer complexes in a 1:1 stoichiometry and of n-sigma type with these molecules. The values of the formation constants Kc of these iodinated complexes indicate a strong donor acceptor interaction. These drugs can therefore be expected to interfere with thyroid metabolism. PMID- 2566393 TI - Agonist-antagonist opioids: theory and clinical practice. PMID- 2566391 TI - Circulatory responses to laryngoscopy: the comparative effects of placebo, fentanyl and esmolol. AB - The circulatory response to a 30-second laryngoscopy followed by orotracheal intubation was recorded in 60 patients of ASA physical status III or IV undergoing a variety of non-cardiac surgical procedures. Patients were randomly allocated to either the placebo, esmolol (500 micrograms.kg-1.min-1 X 6 minutes, followed by 300 micrograms.kg-1.min-1 X 9 minutes), or fentanyl (0.8 microgram.kg 1.min-1 X 10 minutes) group, and the observer was blinded to the infusion administered. Esmolol blunted the heart rate (HR) response, while fentanyl decreased it below the baseline and maintained it there, in spite of laryngoscopy. Similarly, fentanyl decreased the systolic (SBP), mean (MBP) and diastolic blood pressures (DBP) significantly below the baseline, while these pressures were either retained at or elevated slightly above control in the esmolol group. In these doses, the HR response to laryngoscopy was more effectively blocked by fentanyl, while esmolol better retained perfusion pressure. There were no complications or ischaemic electrocardiographic changes in any patient. PMID- 2566392 TI - Cyclosporine-vecuronium interaction. PMID- 2566394 TI - Distribution of beta-adrenergic receptors in failing human myocardium. Implications for mechanisms of down-regulation. AB - The density of beta-adrenergic receptors is reduced in crude membranes prepared from failing human myocardium. We used quantitative autoradiography of radioligand binding sites in intact tissue slices to determine whether the total tissue content of receptors is reduced and to characterize the transmural distribution of receptors in cardiac myocytes and the coronary vasculature in hearts obtained from nine cardiac transplant patients with severe congestive failure. Binding of [125Iodo]cyanopindolol to transmural slices of human myocardium was rapid, saturable, stereoselective, and displaceable by agonists and antagonists with an appropriate rank order of potency. Binding isotherms in four normal and nine failing ventricles showed a significant reduction in the total tissue content of beta-receptors in failing myocardium (38.3 +/- 2.0 fmol/mg protein) compared with normal tissue (52.4 +/- 1.7 fmol/mg protein, p = 0.038). In the normal ventricles, the greatest receptor density was observed autoradiographically in myocytic regions of the subendocardium. Receptor density of the coronary arterioles was approximately 70% of that in adjacent myocytic regions. The density of binding sites in both myocytic regions and arterioles was diminished in all regions of the failing ventricles, but down-regulation was due primarily to a selective reduction of beta-receptors of subendocardial myocytes (63 +/- 5% of subepicardial receptor density vs. 115 +/- 6% in controls, p less than 0.0001). These observations indicate that down-regulation occurs nonuniformly in the transmural distribution and thus is likely not related simply to elevated circulating catecholamine levels. PMID- 2566395 TI - Differences in alpha-adrenergic responsiveness between human internal mammary arteries and saphenous veins. AB - Little is known regarding specific biologic and pharmacologic differences between human internal mammary arteries and saphenous veins. To better define the role of alpha-adrenoceptor-mediated vasoconstriction in human internal mammary arteries and saphenous veins, we obtained fresh specimens of both vessels from 32 patients undergoing coronary artery bypass surgery. Dose-response curves were generated for the relatively selective alpha 1-receptor agonist phenylephrine, the alpha 2 receptor agonist BHT-920, and the alpha 1- and alpha 2-receptor agonist norepinephrine. Phenylephrine elicited similar contractile responses in internal mammary arteries and saphenous veins, with a mean EC50 (the effective concentration necessary to produce 50% of the maximal contraction) of 1.4 X 10( 6) M for internal mammary arteries and 1.8 X 10(-6) M for saphenous veins (p = NS). Selective stimulation of alpha 2-receptors with BHT-920 elicited a marked contractile response only in saphenous veins. Dose-response curves for phenylephrine and BHT-920 were shifted to the right for both vessels in the presence of the alpha 1-receptor antagonist prazosin and the alpha 2-receptor antagonist yohimbine, respectively. Norepinephrine elicited contraction at a lower concentration in saphenous veins than in internal mammary arteries with a mean EC50 of 7.8 X 10(-8) M for saphenous veins and a mean EC50 of 3.4 X 10(-7) M for internal mammary arteries (p less than 0.05). The results suggest that alpha adrenoceptor-mediated vasoconstriction is caused primarily by alpha 1-receptors in human internal mammary arteries and by alpha 1- and alpha 2-receptors in human saphenous veins. PMID- 2566396 TI - The development of N-methyl-D-aspartate receptors in cat visual cortex. AB - During a critical period of early postnatal development, the visual cortex of kittens is susceptible to experience-dependent modifications of neuronal response properties. Recently, the activation of N-methyl-D-aspartate (NMDA) receptors has been identified as an indispensable prerequisite for the induction of such modifications. We therefore investigated developmental changes in the density and distribution of NMDA receptors and questioned whether these showed a relation to the time course of the critical period. We determined the proportion of [3H]glutamate binding sites that were displaced by the NMDA receptor antagonist 2 amino-5-phosphonovalerate (APV) on 10-microns-thick cryostat sections of the primary visual cortex. The overall density of APV-sensitive [3H]glutamate binding sites increased dramatically between the second and the fourth week and stayed at this level throughout the critical period. Towards the end of the critical period, these binding sites decreased and finally reached adult values that were slightly above those of 2-week-old kittens. APV-sensitive binding sites were present in all cortical layers of the age groups investigated. While the general pattern of developmental changes was similar in all layers, slight differences existed in the time course. These observations are compatible with the notion that NMDA receptor activation is required for the expression of use-dependent change of response properties in the kitten visual cortex. Furthermore, they suggest as a possible reason for the decline of malleability towards the end of the critical period the reduction of NMDA receptors. PMID- 2566397 TI - Persistent hypotensive effect of L-dopa given early during development to rats with inherited stress-induced arterial hypertension. AB - A long-lasting decrease of the basal and stress-induced arterial blood pressure was obtained in rats with inherited emotional stress-induced arterial hypertension by means of injections of the dopamine precursor L-DOPA during early development (21-25 days after birth). The restoring effect of L-DOPA was produced through enhancement of synthesis of the brain noradrenaline and, perhaps, adrenaline. The effect was associated with a normalization of the response of the brain adrenergic system to noradrenaline and, presumably, with increase of tyrosine hydroxylase activity in the cortex and hindbrain. PMID- 2566398 TI - Activation energy and lectin affinity chromatography of gamma-glutamyltransferase as a marker for enzyme heterogeneity. AB - Lectin affinity chromatography of gamma-glutamyl transferase (GGT,EC 2.3.2.2) is able to detect differences in the carbohydrate moiety of the enzyme. Binding of tissue GGT towards lectins is significantly different from serum GGT, showing increased galactosylation in tissue forms. Kidney GGT is less glycosylated than GGT from other tissues (liver, pancreas, prostate, vesiculae seminales). Increases in sialic acid content of GGT are associated with an increase in the activation energy of the catalyzed reaction. Differences in galactose, fucose and N-acetylhexosamine content induce much smaller effects on activation energy. In liver diseases, serum GGT is characterized by an altered affinity against lectins recognizing galactose, fucose and N-acetyglucosamine and by increased activation energy. In patients with liver disease, use of fixed temperature conversion factors can lead to erroneous calculations of serum GGT enzyme activity (errors up to 13.3%). PMID- 2566399 TI - A simplified and rapid test for acetylator phenotyping by use of the peak height ratio of two urinary caffeine metabolites. AB - We describe a simplified liquid-chromatographic test in which acetylator phenotype is determined by measuring the peak height ratio of two urinary caffeine metabolites, 5-acetylamino-6-formylamino-3-methyluracil and 1 methylxanthine. We applied this test to determine the acetylator phenotypes of 52 subjects who regularly drink coffee, tea, or caffeinated beverages. Also, we determined the acetylator phenotypes of these subjects according to a well established sulfasalazine test, which yielded identical results. We established the reproducibility of the described test by determining the acetylator phenotypes of 10 additional subjects on two different days separated by a period of two to five weeks. Of the 52 subjects examined by both tests, 40 (76.9%) were classified as slow acetylators, which agrees well with the percentage reported elsewhere for 297 similar subjects from the Saudi population. PMID- 2566400 TI - Circadian variations and reference intervals for some enzymes in urine of healthy children. AB - Circadian variations of alanine aminopeptidase (EC 3.4.11.2), gamma glutamyltransferase (EC 2.3.2.2), and N-acetyl-beta-glucosaminidase (EC 3.2.1.30) in urine were studied in 10 healthy children, ages six to 15 years. Urine specimens were collected during 24 h, grouped into four time intervals. Enzymes were measured spectrophotometrically, with automation. These enzymes all showed diurnal variation, with morning (8 a.m.-12 noon) excretion being highest. We also analyzed timed urinary specimens (8 a.m.-12 noon) from 136 healthy children, ages two to 11 years. Reference intervals are presented for these enzymes. High excretion of the three enzymes was observed in children two and three years old. PMID- 2566402 TI - Surgery for peptic ulcer disease in the era of H2 receptor blockers. PMID- 2566401 TI - The serum activities of AP, gamma-GT, GLDH, GPT and CHE after complete biliary obstruction and choledochocaval fistula in the rat. AB - The activities in serum of alkaline phosphatase, gamma-glutamyltransferase, glutamate dehydrogenase, glutamic pyruvic transaminase and cholinesterase were compared after complete biliary obstruction (CBO) and choledochocaval fistula (CCF) in the rat. CCF was used as a model of complete biliary retention without bile stasis and without increased pressure in the biliary tract. The increases in AP, GLDH and gamma-GT within 24-h post-op. show no difference between the two experimental groups. The conclusion is that the retention of biliary constituents alone is responsible for the increase in the levels of serum activity and that other conditions like bile stasis and increased pressure in the biliary tract do not play an important role in the pathogenesis of these alterations. The rise of GPT activity in CCF is of a lesser degree than in CBO. PMID- 2566403 TI - Genetic heterogeneity between two clinical forms of cystic fibrosis evidenced by familial analysis and linked DNA probes. AB - CF heterogeneity has been evidenced from both clinical and genetic observations. At least two clinical forms of CF are easily distinguishable: CF with meconium ileus and CF without meconium ileus. The results of prenatal diagnosis have shown that the recurrence rates of CF are different in these two clinical forms. Molecular analysis of Restriction Fragment Length Polymorphisms (RFLPs) tightly linked to the cystic fibrosis (CF) gene defined several types of CF and normal chromosomes in a French sample of 64 families with CF. The CF mutation is tightly linked to one XV-2C and KM19 RFLPs haplotype but is differently linked to J3.11 RFLP alleles, depending on whether or not the clinical form of CF is associated with ileus. A distortion of the segregation ratio observed between normal and CF haplotypes in the families with ileus could explain the high recurrence rate of CF in such families. PMID- 2566404 TI - Early and late posttraumatic foot reconstruction. AB - When complex trauma occurs in the foot, there is (1) loss of structural continuity that renders the bony architecture mechanically useless and (2) changes in the soft tissue that subject the foot to potential contractures and atrophy. The priorities of salvage of such injuries are early and accurate diagnosis, early reduction and stabilization, careful handling of soft tissues, tissue transfer to fill defects, and early motion. By these means it is possible to diminish the effects of edema, atrophy, stiffness, and osteoporosis. When initial treatment fails, salvage procedures must consider the length of disability, the expected functional outcome after salvage, and the lifestyle of the individual. Late salvage is by selective arthrodesis and realignment of mechanical relationships, followed by aggressive functional rehabilitation. PMID- 2566405 TI - Effects of sodium depletion on renal prostanoid synthesis in rats: influence of the converting enzyme inhibitor captopril. AB - 1. The synthesis of prostaglandin (PG) E2, PGF2 alpha, 6-keto-PGF1 alpha and thromboxane (TX) B2 by isolated glomeruli, cortical tubules, inner medullary slices and outer medullary slices was measured in salt-depleted (LNa) rats and in salt-depleted rats receiving captopril (LNa-CEI). Animals were studied before and after 4, 9 and 15 days of Na+ depletion. 2. Na+ balance was reached in LNa rats after 4 days. Blood pressure and creatinine clearance remained stable. Serum Na+ decreased from 140 +/- 1 to 126 +/- 1 mmol/l (mean +/- SEM, P less than 0.01). In contrast, LNa-CEI rats were unable to conserve Na+ adequately: fractional excretion of Na+ and natriuresis were constantly greater than in LNa animals. As a consequence, LNa-CEI rats developed severe hyponatraemia, lost weight and their creatinine clearance decreased. 3. The glomerular synthesis of PGE2, PGF2 alpha and 6-keto-PGF1 alpha, but not of TXB2, was significantly increased in LNa rats. In LNa-CEI rats, the synthesis of PGE2 and 6-keto-PGF1 alpha was similar to control values, but PGF2 alpha and TXB2 synthesis was elevated at day 9. In cortical tubules, PGE2 and PGF2 alpha were unaffected by Na+ depletion, but 6 keto-PGF1 alpha and TXB2 were increased and a similar trend was observed in LNa CEI rats. In outer medulla of LNa rats, a decrease in all the eicosanoids measured was observed at day 4. In LNa-CEI animals, the synthesis of PGE2 and PGF2 alpha, but not of 6-keto-PGF1 alpha and TXB2, was significantly depressed. In inner medulla, Na+ depletion only tended to decrease PGF2 alpha and 6-keto PGF1 alpha, but in the presence of captopril, the synthesis of all prostanoids was significantly decreased. PMID- 2566406 TI - [Therapy of insomnia]. AB - The authors review the neurochemical and electrophysiological features of insomnia, together with the results obtained by various substances. The literature data show that the benzodiazepines (BZ) should be administered for short periods of time, in order to avoid addiction and withdrawal symptoms. For this reason, the authors suggest that, before starting a therapy with such substances, an accurate clinical evaluation should be made and a good knowledge of the pharmacokinetics of the various BZ is essential. PMID- 2566407 TI - Enzymatic influences on amino acid transport across the small intestine. PMID- 2566408 TI - Cross-adapted sugar responses in the mouse taste cell. AB - 1. Intracellular recordings of mouse taste cell responses were made using a glass micro-electrode filled with Procion yellow dye solution. 2. Six sugars (sucrose, maltose, lactose, glucose, galactose and fructose) produced the depolarization responses. 3. Gustatory cross adaptation between sugars was determined. When the taste cell was pre-adapted with one of the six sugars, the other five sugars, cross adapted, produced depolarization, hyperpolarization or null responses. 4. From these observations, it is suggested that there are multiple sugar receptor sites on the receptor membrane of the mouse taste cell. PMID- 2566410 TI - Porcine adrenal prolactin receptors: characterization, changes during neonatal development and effects of hypoprolactinemia. AB - 1. Adrenal prolactin (PRL) receptors were identified within the adrenal cortex of pigs (Sus domesticus), and found to be located specifically on isolated zona fasciculata/reticularis cells (6437 sites per cell). 2. These PRL receptors were associated with binding to [125I]-oPRL which was characterized as being time and temperature dependent, specific for PRL, saturable, of high affinity (Ka = 10(10)/M) with a single class of binding sites, and irreversible except under extreme conditions. 3. The concentrations (fmol/mg protein) of PRL receptors decreased by 35% (P less than 0.05) between 3 and 10 days of age, and subsequently remained constant until 30 days of age. Total content (fmol/paired adrenals) increased progressively (2-fold, P less than 0.05) between 3 and 30 days of age. 4. Short-term (less than 16 hr) and prolonged (7 weeks) hypoprolactinemia (46-64% of control levels, P less than 0.05) were not associated with changes in numbers of porcine adrenal unoccupied PRL receptors. PMID- 2566409 TI - Selective procaine inhibition of rat chorda tympani responses to electric taste stimulation. AB - 1. The lingual treatment of 1% procaine for 10 min selectively suppressed responses of the rat chorda tympani nerve to anodal current applied to the tongue with NaCl in the bathing medium to about 50% of control but the drug produced no significant suppression in responses to chemical taste stimuli. 2. The magnitude of suppression of response to anodal current varied with concentration of procaine and kind of bathing medium for the current stimulation (larger in the order of NaCl greater than KCl greater than CaCl2 greater than HCl). 3. Such ion specificity in procaine suppression suggests that responses of the chorda tympani nerve to anodal current are provoked through the taste cell (not direct action on the taste nerve), and that the receptor mechanisms for anodal current are at least partly different from that for chemical taste stimuli. PMID- 2566411 TI - Body and egg iron content in shaver and New Hampshire chickens. AB - 1. A comparative study on the iron content in internal organs (blood, liver, spleen, kidneys and heart), carcass and feathers, as well as on the whole iron content has been carried out at different ages (4, 8, 13, 18 and 24 weeks), on two chicken breeds (shaver and New Hampshire). 2. The iron content in internal organs in shaver was higher than in New Hampshire. 3. A tendency of the iron content in the carcass to decrease with age has been observed for both breeds; values were higher for laying shaver hens. 4. The egg-producing strain, shaver, showed a greater iron status with a stable body iron content from 13 weeks on (53 55 ppm), permitting a constant laying frequency (90%) during an 18-month period, with only a small reduction of egg iron content from 33.8 to 31 ppm. 5. The whole body iron decreased progressively with age in the New Hampshire broiler strain to 42-49 ppm iron, and the laying process sharpened this tendency. A drastic reduction in laying, accompanied by a lower egg iron content (28.9 ppm iron), was observed. 6. Iron data obtained in the present paper reflect the specific genetic adaptation to egg production of the shaver strain. PMID- 2566412 TI - Total body haematocrit iron kinetics and erythrocyte life span in pigeons (Columba livia). AB - 1. The mean pigeon erythrocyte life span was found to be 17-25 days by Cr51 labeled erythrocytes and 21 +/- 3.4 days by iron kinetics. 2. Total red blood cell volume has been calculated by Cr51-labeled erythrocytes while total plasma volume was determined both by a dye method and iron kinetic data. From these results total blood volume and total body haematocrit were found to be 0.090 +/- 0.002 ml/g body wt and 36 +/- 4.3%, respectively. 3. Venous haematocrit, haemoglobin concentration, erythrocyte count, mean corpuscular haemoglobin concentration, plasma iron and red blood cell iron have also been measured. 4. A significant difference between total body and venous haematocrit and a short mean red blood cell life span, due to ageing and to random destruction of erythrocytes were shown. 5. The above observations are compared with analogous available data for human beings and their physiological significance is discussed. PMID- 2566413 TI - Ventilatory responses of the ground squirrel, Spermophilus tridecemlineatus, to various levels of hypoxia. AB - 1. Minute ventilation (VE) in the semifossorial ground squirrel (Spermophilus tridecemlineatus) increased with increased levels of hypoxia. 2. The increase in VE was brought about primarily by an increase in breathing frequency (f). There was no significant change in tidal volume (VT). 3. The PiO2 threshold for the ventilatory response to hypoxia and position of the ventilatory response curve in the ground squirrel were closer to the semifossorial echidna (Tachyglossus aculeatus) than the completely fossorial mole rate (Spalax ehrenbergi); both the ground squirrel and echidna had a higher PiO2 threshold than the mole rat. 4. The ventilatory response curve was shifted to the left in the mole rat. 5. These observations indicate that the mole rat is the least responsive to hypoxia of the three species. PMID- 2566414 TI - The change in the rate of muscle protein metabolism of rats from weaning to 90 days of age. AB - 1. Protein synthesis, net deposition and breakdown was studied in the gastrocnemius muscles of growing rats between weaning and 90 days of age. 2. Fractional protein synthetic rates declined from 30.02% at 25 days to 7.41% at 90 days. 3. The rate of protein degradation follows a similar pattern to that of protein synthesis. A linear relationship was found. 4. The break in the growth curve between 30 and 31 days was also observed in protein metabolism. PMID- 2566415 TI - Urinary immunoreactive androgen levels during sexual development in the male tree shrew (Tupaia belangeri). AB - 1. Immunoreactive androgens were measured in the urine of individual male tree shrews throughout post-natal development. 2. Urinary androgens were low during the infantile phase and then rose significantly between the 36-45 and 46-55 day age groups, in association with the onset of pubertal development. 3. Androgen excretion increased linearly during the pubertal period in parallel with the progressive development of the testes and reproductive tract. 4. The precise endocrine correlates established in the present work suggest that serial determination of urinary androgens provides a reliable method of monitoring male reproductive development which may be applied to small, sensitive species. PMID- 2566416 TI - Salivation in the red kangaroo (Macropus rufus) during sympathetic nerve stimulation. AB - 1. Continuous electrical stimulation at low frequency (5 Hz) and short pulse duration (500 microseconds) of the cervical sympathetic trunk for periods up to 15 min caused no obvious flow from the parotid or mandibular glands of the red kangaroo. 2. Higher frequencies combined with longer pulse durations caused both glands to secrete. Flow reached maximum in less than 3 min and then declined but, on cessation of stimulation, flow increased again for a short period. This flow response may be caused by the interaction of the secretory response with myoepithelial contraction. 3. The parotid saliva had substantially higher protein, phosphate and hydrogen ion concentrations, and lower sodium concentrations than cholinergic parotid saliva. The low pH indicates bicarbonate concentrations far lower than in other sympathetic salivas. 4. The mandibular saliva had higher protein, urea and potassium, and lower chloride and hydrogen concentrations than cholinergic mandibular saliva. PMID- 2566417 TI - Seasonal levels of cortisol, triiodothyronine and thyroxine in male axis deer. AB - 1. Seasonal plasma levels of thyroxine (T4), triiodothyronine (T3) and cortisol were investigated between November and June in seven penned male Axis deer. 2. No distinct seasonal variation of cortisol has been detected. The levels oscillated between 1 and 5 micrograms/dl. 3. The stress of immobilization and sampling had little effect on cortisol levels. Concentrations remained mostly stable in three consecutive samples taken 10 min apart. 4. T3 concentrations were stable between November and March (average values 110-120 ng/dl). After a sharp decline in April (average 70 ng/dl), a strong rebound in May and June was observed. 5. A distinct seasonal peak of T4 (highest individual value, 12.1 micrograms/dl) was detected in March. After a sharp decline in April (lowest individual value, 4.5 micrograms/dl) a strong rebound followed in May. PMID- 2566419 TI - Variation in renal structure and urine concentrating capacity among ground squirrels of the Spermophilus townsendii complex (Rodentia: Sciuridae). AB - Renal structure and urine concentrating capacity varied significantly among nine populations of desert ground squirrels representing four closely related taxa. Variation in renal structure was weakly associated with taxonomic grouping. Urine concentrating capacity was not correlated with standard measures of habitat were aridity, but it was correlated with soil salinity. Maximum urine osmolalities for one population from the most saline habitat were among the highest documented for sciurid rodents. Results do not support the hypothesis that hibernating sciurids have poor renal efficiency. PMID- 2566418 TI - Rate of water turnover and electrolyte balance of an arid-zone marsupial, the spectacled hare wallaby (Lagorchestes conspicillatus) on Barrow Island. AB - The work reported in this paper describes aspects of the water and electrolyte metabolism of free-ranging spectacled hare wallabies (Lagorchestes conspicillatus) on Barrow Island in Western Australia. Two populations were studied in both spring (October) and summer (March). Rates of water turnover were measured in spring with tritiated water and were extreme ly low, approximating 5% of the total body water per day, and these are amongst the lowest rates yet published for any marsupial. Urine outputs were also extremely low and variable in spring, and urine osmolality averaged from 859 to 1015 mOsm/kg. In summer, rates of urine production were significantly higher, but urine osmolality did not change. Osmolar clearance was elevated in animals collected in March, and free water clearance was more negative but, in view of higher rates of urine production, a significant modification of renal function is not indicated. Previous surveys have indicated that the density of hare wallabies is greater in disturbed areas of Barrow Island. Dietary analysis suggests that this unequal distribution results from a greater availability of preferred plant species growing in these disturbed areas. PMID- 2566421 TI - The effect of food restriction on circulating insulin-like growth factor-I in mice divergently selected for high or low protein or fat to body mass ratios. AB - 1. Plasma insulin-like growth factor-I (IGF-I) was measured in mice divergently selected for high and low lean tissue gain or high and low fat to body weight ratio, both before and after fasting and refeeding. 2. Selection for high lean tissue resulted in increased body weight and a higher basal IGF-I concentration at 10 weeks of age. 3. Selection resulting in a difference in fatness had no effect on IGF-I concentration. 4. Circulating IGF-I decreased more rapidly in response to 24 hr food withdrawal in growing (5-week-old) than in almost fully grown (10-week-old) animals from all lines. PMID- 2566422 TI - Sodium and potassium concentrations in the intestinal fluids of the fowl, Gallus domesticus. AB - 1. Na and K concentrations in the luminal fluids on the jejunum, ileum and colon were measured in domestic fowl on diets containing different amounts of Na and K. 2. Physiological adjustments of the Na and K content of these fluids were observed to occur in all three intestinal segments. 3. Regulation of gastrointestinal losses of Na and K appears to be initiated in the anterior regions of the intestines of the domestic fowl and is maintained or amplified as the ingesta moves posteriorly. PMID- 2566423 TI - Hyperoxia decreases lung size of amphibian tadpoles without changing GSH peroxidases or tissue peroxidation. AB - 1. During the development of D. pictus larvae (Amphibia) in normoxia, selenium (Se) GSH-Px increased whereas non-Se GSH-Px did not change. 2. Acclimation to 60 or 100% O2 did not change Se GSH-Px or non-Se GSH-Px. 3. Hyperoxia did not change tissue peroxidation (TBA-RS) confirming the good capacity of D. pictus tadpoles for O2-adaptation. 4. Since hyperoxic induction of catalase (CAT) has been previously described in D. pictus tadpoles, it is concluded that CAT is more important than both GSH-Px for the establishment of O2-adaptation. 5. Increases of Se GSH-Px, SOD and CAT, are probably important for adaptation to the change from aquatic to aerial environment during metamorphosis in normoxia. 6. Chronic exposure to 100% O2 enormously reduced the lung size of D. pictus larvae. PMID- 2566420 TI - An in vitro preparation of the extensor digitorum communis muscle from the chick (Gallus domesticus) for studies of protein turnover. AB - 1. Extensor digitorum communis (EDC) muscles from the chick synthesized and degraded proteins at linear rates, and maintained high levels of ATP, creatine phosphate and glycogen, for 9 hr of incubation. 2. The apparent viability of the EDC preparation was improved by the provision of a continuous supply of oxygen, glucose, insulin and amino acids at the levels found in chick plasma, and incubation at 33.5 degrees C. 3. Incubated muscles were in net negative protein balance; however, under optimal conditions, this represented only 0.03% of total protein/hr. Rates of protein synthesis in the EDC preparation were 65% of those determined in vivo, comparing favourably with values obtained by other workers in mammalian muscle. 4. Rates of protein synthesis responded to insulin, amino acids, and to the nutritional status of the bird, as had been observed in prior studies with mammalian muscle. By contrast to mammalian muscle protein degradation was relatively insensitive to these modifiers. PMID- 2566424 TI - Differences in the protein composition of rat parotid salivas evoked by sympathetic and parasympathetic nerve stimulation. AB - 1. Parotid salivas were collected from rats following electrical stimulation of either the sympathetic or parasympathetic nervous supplies. 2. The protein compositions of these salivas were compared using high performance ion-exchange chromatography (FPLC), SDS electrophoresis and biochemical assay. 3. Chromatography and electrophoresis indicated differences between the protein compositions of sympathetic and parasympathetic salivas. 4. Protein secretion derived solely from the exocytosis of parotid acinar cells cannot account for these differences. PMID- 2566425 TI - Hibernation in a monotreme, the echidna (Tachyglossus aculeatus). AB - The body temperatures of five echidnas in Australia's Southern Alps were monitored by radio telemetry from February to December 1987. All five hibernated throughout the winter, showing very low body temperatures (4-9 degrees C, close to ambient) when torpid, compared with 28-33 degrees C in a typical day during the active season. Spontaneous arousals from hibernation occurred every 2-3 weeks, during which body temperatures rose rapidly to over 30 degrees C for several hours before dropping to be close to ambient again. The identification of "classical" hibernation in a monotreme, with a similar pattern to that seen in Eutheria and in an animal as large as the largest eutherian hibernator, has important implications for current ideas about the evolution of endothermy. PMID- 2566426 TI - Metabolic responses of chicken embryos to graded, prolonged alterations in ambient temperature. AB - 1. Chicken embryos aged 12, 16, 18 and 20 (externally pipped) days of incubation were exposed to graded reductions (2 degrees C) in ambient temperature from 38 to 28 degrees C, exposure to each temperature lasting up to 9 hr. 2. Oxygen uptake was measured first at 38 degrees C and then in the quasi-equilibrium state at lowered temperatures. The temperature coefficient (Q10) was calculated for each egg. 3. For mild cooling (32 degrees C), the Q10 in 18-day-old embryos was about 1.5, while 12- and 16-day-old embryos had a Q10 value of about 2, indicating that a feeble homeothermic metabolic response to cooling appears in late prenatal embryos. It became more marked in externally pipped embryos and further augmented in hatchlings. PMID- 2566427 TI - Metabolic responses to gradual cooling in chicken eggs treated with thiourea and oxygen. AB - 1. Late prenatal chicken embryos in eggs injected with saline showed a feeble homeothermic metabolic response to gradual cooling. This response was absent in thiourea-treated eggs. This suggests that the incipient homeothermic metabolic response before paranatal life may be attributed to thyroid development. 2. The compensatory metabolic response disappeared in embryos exposed to a hypoxic environment, while it was augmented in eggs in pure O2, decreasing as ambient temperature fell. 3. These results may indicate that the homeothermic metabolic response in late embryos is O2-conductance-limited and power-limited as previously suggested. PMID- 2566428 TI - Monoaminergic nerves in the skin of plaice Pleuronectes platessa (L.). AB - 1. The fluorescent histochemical technique of Falck and Hillarp was applied to plaice skin. The presence of monoaminergic nerve terminals, containing predominantly stores of adrenaline, forming a plexus in and around the melanophore layer was demonstrated. 2. Such stores were enriched by noradrenaline in the presence of monoamine oxidase inhibitor, unaffected by spinal section, depleted by spinal nerve section or ligatures and abolished by reserpine. 3. The observations support the view that teleost sympathetic melanophore aggregating nerves are truly adrenergic. PMID- 2566429 TI - Gastrin/CCK-like peptides in the spiny dogfish, Squalus acanthias; concentrations and actions in the gut. AB - 1. The presence of gastrin/CCK-like immunoreactive material in both muscle and mucosal layers of the whole gut, except the cardiac stomach, in Squalus acanthias has been confirmed by radioimmunoassay. The highest levels were measured in rectum and spiral intestine respectively. 2. Fractionation of the spiral intestine mucosal extract on DEAE 52 columns indicated the presence of multiple forms of gastrin/CCK in the elasmobranch gut. 3. Synthetic mammalian gastrin- and CCK-like peptides, when effective, increased the rhythmic activity and sometimes the basal tonus in smooth muscle preparations from the intestine or rectum. 4. The irregular effects of mammalian peptides on the motility indicated an essential difference from the mammalian counterpart in the structure of the elasmobranch peptide present in the gastrointestinal nerves. PMID- 2566430 TI - Effects of 2-arylbenzimidazoles on rat hepatic microsomal monooxygenase system. AB - 1. The effects of eight newly synthesized 2-aryl substituted benzimidazole derivatives on control and phenobarbital (PB) treated rat liver microsomal aniline 4-hydroxylase and ethylmorphine N-demethylase activities, and their binding to control and PB-treated rat liver microsomal oxidized cytochrome P-450 are presented. 2. All compounds inhibited ethylmorphine N-demethylase activity with I50 values ranging from 8.50 x 10(-4) M to 27.83 x 10(-4) M in control and ranging from 2.80 x 10(-4) M to 15.79 x 10(-4) M in PB-treated rats. 3. Aniline 4 hydroxylase activity was inhibited by all of the compounds tested having I50 values in the range of 7.04 x 10(-4) M-31.37 x 10(-4) M in PB-treated rats, but only five of the compounds showed inhibitory activity in control rats. 4. Only a few significant regression coefficients could be found between the parameters of the chemicals studied and their inhibitory patterns. 5. No correlation has been observed between the binding of the derivatives and their inhibitory pattern. PMID- 2566431 TI - Kinins in ant venoms--a comparison with venoms of related Hymenoptera. AB - 1. Venom preparations have been made of six ant, one pompilid wasp, two mutillid wasp, and four social wasp species. 2. The venoms were analysed pharmacologically in order to detect kinin-like activity. 3. Due to the small amounts of venoms available only a cascade of smooth muscle preparation could be used. 4. Kinin activities have been found in five ant venoms and in four social wasp venoms. 5. No kinin activity has been found in the venoms of the pompilid and mutillid wasps. 6. All ant venoms also contain unidentified agonists for vertebrate smooth muscle preparations. PMID- 2566432 TI - Effects of tributyltin compounds on ionic regulation and gill ATPase activity in estuarine fish. AB - 1. Striped bass (Morone saxatilis) were exposed to 0, 0.10, 0.34, or 1.09 micrograms/l tributyltin (TBT) for 14 days. Gill Mg2+ ATPase and serum Na+, K+, Ca2+ and Mg2+ were not altered. Na+K+ATPase was significantly (P less than 0.05) increased (+48%) at 0.10 microgram/l. 2. In striped bass gill homogenates exposed to TBT in vitro, there was significant inhibition of Na+K+ATPase at 106.0 micrograms/l (17.2%) and Mg2+ATPase at 53.0 and 106.0 micrograms/l (16.0% and 24.3%, respectively). 3. In mummichog (Fundulus heteroclitus) gill homogenates, there was significant inhibition of Na+K+ATPase at 25.3 and 50.5 micrograms/l (10.9% and 16.1%) and Mg2+ATPase at 5.1, 25.3, and 50.5 micrograms/l (26.7%, 32.2%, and 36.2%). PMID- 2566433 TI - Synthetic egg-laying hormone of Aplysia: quantitative studies of induction of egg laying in Stylocheilus and of the activation of Aplysia buccal neuron B16. AB - 1. Synthetic Aplysia egg-laying hormone (ELH-lysine-amide) elicited egg laying in Stylocheilus at a threshold dose of 0.5 microgram per recipient, estimated to be a concentration in the circulation of Stylocheilus of approximately 70 nM. 2. Threshold level and size of egg mass produced by ELH-lysine-amide and bag cell extracts (containing biological ELH) were not significantly different. Latency to lay of recipients of 0.5-4.0 micrograms ELH-lysine-amide (30 +/- 1 min) was significantly longer (P less than 0.05) than for bag cell extract recipients (21 +/- 1 min). 3. ELH-lysine-amide depolarized and activated action potentials in Aplysia buccal neuron B16 in high magnesium, low calcium medium. 4. The lowest concentration of ELH-lysine-amide to activate a supra-threshold response of left and right B16 neurons ranged from 250 nM to 1 microM. 5. Threshold levels for responses to synthetic ELH-lysine-amide and to biological ELH were approximately the same in both egg-laying assay and electrophysiological assay, indicating the likely identity of synthetic and biological ELH. However, the shorter egg-laying latency with bag cell extract suggests that there may be additional factors in the extract that facilitate egg laying. PMID- 2566434 TI - Effect of continuous and intermittent clenbuterol feeding on rat growth rate and muscle. AB - 1. The growth response to clenbuterol is a dynamic process. 2. Body weight gain is stimulated within two days of treatment and the effect attenuates by two weeks of treatment. 3. Intermittent feeding prevents the attenuation of the growth response. 4. Muscle weight increased 14-22% by both feeding regimens. 5. Clenbuterol decreased cathepsin B activity in the EDL and gastrocnemius and increased the activity in the soleus after two weeks of continuous clenbuterol treatment. PMID- 2566435 TI - Phagocytosis in Tetrahymena thermophila: naloxone-reversible inhibition by opiates. AB - 1. Nanomolar concentrations of opiates inhibit phagocytosis in the ciliated protozoan Tetrahymena thermophila. 2. Naloxone and naltrexone counteract the effect of the opiate agonists tested. 3. The dose-response curves are U-shaped, with no detectable effect at low or high concentrations. 4. An increase in extracellular calcium and dopamine counteract the inhibition caused by metenkephalin. 5. The recognition mechanism for opiates in Tetrahymena cannot be classified as belonging to any of the mammalian opiate receptor subtypes and is perhaps a primitive receptor. PMID- 2566436 TI - Histamine directly acts on beta-adrenoceptors as well as H1-histaminergic receptors, and causes positive inotropic effects in isolated ventricular muscles of carp heart (Cyprinus carpio). AB - 1. The mechanism for positive and negative inotropic effects of histamine was studied in electrically stimulated ventricular strips of carp heart. 2. A high concentration of histamine (1 mM) caused a transient negative, and subsequent positive inotropic effects. The positive effect was significantly reduced by pyrilamine, diphenhydramine or dl-propranolol, but was not affected by cimetidine or d-propranolol. 3. Prior treatment with reserpine significantly decreased epinephrine and norepinephrine contents in ventricular muscles, and also almost completely abolished the positive inotropic effect caused by tyramine; however, this treatment failed to affect the positive inotropic effect of histamine. 4. The transient negative inotropic effect was reduced by neither atropine, diphenhydramine, pyrilamine nor cimetidine, and potentiated by pyrilamine. 5. These results suggest that the positive inotropic effect of histamine observed in the ventricular muscle of carp heart is mediated by a direct stimulation of both H1-receptors and beta-adrenoceptors. The negative inotropic effect is unrelated to either cholinergic or histaminergic receptor stimulation. PMID- 2566437 TI - The roles of alpha- and beta-adrenoceptors in the chronotropic responses to norepinephrine in carp heart (Cyprinus carpio). AB - 1. The chronotropic effect of norepinephrine was studied in isolated spontaneously beating atrial preparations of carp (Cyprinus carpio) heart. 2. Norepinephrine, 0.1 microM, caused a positive chronotropic effect, while at 1 microM it caused either a positive or a negative chronotropic effect. The positive chronotropic effect, observed in 13 preparations, was potentiated by phentolamine and almost completely blocked by propranolol. 3. The negative chronotropic effect observed in the other 5 preparations was greater in the presence of propranolol, reduced by phentolamine and not affected by atropine. 4. These results indicate that alpha- and beta-adrenoceptors may coexist, mediating the negative and positive chronotropic effects, respectively, in isolated atrial preparations of carp heart. PMID- 2566438 TI - Ca2+ dependence of inositol 1,4,5-trisphosphate 3-kinase activity in the cytosol fraction of pig coronary artery. AB - 1. The activity of inositol 1,4,5-trisphosphate 3-kinase in subcellular fractions of smooth muscles of the pig coronary artery was examined. 2. Incubation of [3H]inositol 1,4,5-trisphosphate (IP3) with muscle homogenates produced more polar 3H-radioactivity (probably as inositol 1,3,4,5-tetrakisphosphate, IP4) than IP3, in the Mg2+- and ATP-dependent manner, thereby indicating the presence of IP3 3-kinase activity in homogenates of the muscle. 3. Most of the kinase activity was present in the cytosol fraction. The enzyme activity was reversibly activated by Ca2+ with a half-maximal effective concentration of 2.5 x 10(-7) M. 4. The calmodulin antagonists, W-7 and chlorpromazine inhibited the Ca2+ activated enzyme activity. PMID- 2566440 TI - A system for the application of general anaesthetics and other volatile agents to superfused, isolated tissue preparations. AB - 1. A delivery system for the application of general anaesthetics or other gaseous and volatile agents to superfused, isolated preparations is described in detail. 2. This system delivers known concentrations of anaesthetic and controls for evaporation and absorption of volatile agents, whilst allowing intracellular electrophysiological recordings to be made from the tissue with minimal disturbance. 3. In particular, this delivery system permits accurate, controlled experiments to be carried out on the neuronal actions of general anaesthetics. PMID- 2566439 TI - A comparison of the actions of 4-aminopyridine, caffeine and quinine on the toad isolated rectus abdominis muscle. AB - 1. 4-Aminopyridine (4-AP)-induced contractures have been compared with those evoked by caffeine and quinine on the toad rectus abdominis muscle. 2. All three compounds produced slowly-developing sustained contractures. The time to half maximal contracture and relaxation was significantly longer for 4-AP than for caffeine or quinine. 3. Verapamil and manganese inhibited 4-AP, caffeine and quinine-induced contractures. 4. Ca2+-free-EGTA Ringer and procaine severely inhibited caffeine and quinine responses, but 4-AP contractures were relatively unaffected. 5. In depolarizing (100 mM K+) Ringer solution, caffeine and quinine responses were reduced to 6-9% of their controls. 4-AP responses were reduced by about 25%. 6. It is concluded that in the toad rectus muscle, 4-AP-induced contractures differ from those produced by caffeine and quinine, and appear to rely mainly on the release of intracellular located Ca2+, while caffeine and quinine are considered to act predominantly on plasma membrane sites. PMID- 2566441 TI - The actions of three volatile general anaesthetics on withdrawal responses of the pond-snail Lymnaea stagnalis (L.). AB - 1. The gastropod mollusc Lymnaea stagnalis (L.) is an ideal model system for studies on anaesthesia. It is reversibly anaesthetized by the general anaesthetics halothane, enflurane and isoflurane. 2. Criteria for "anaesthesia" in Lymnaea were established. The reflex used in ED50 trials was the whole animal withdrawal reflex. 3. ED50 values for halothane, enflurane and isoflurane were, 0.83% v.v. (volume for volume), 1.01% v.v. and 1.09% v.v. respectively. 4. Relationships between anaesthetic concentrations, weights of animals and mortality rates are reported. PMID- 2566442 TI - In vitro inhibition of methionine incorporation in the dorsal bodies of Helix aspersa by synthetic FMRFamide. AB - 1. In in vitro conditions, synthetic FMRFamide was shown to inhibit the uptake of labelled methionine by the dorsal bodies (DB)-containing connective tissue of Helix aspersa. 2. This effect occurred at physiological concentrations and in a dose-dependent manner. 3. Among the different cell types of the explants, the DB cells preferentially incorporated the radioactive precursor. 4. These results suggest that FMRFamide plays a biological role in controlling the DB activity (protein synthesis) of Helix aspersa. PMID- 2566443 TI - The effect of MS 222 an anaesthetic on the peroxide metabolism enzymes in erythrocytes of freshwater and marine fish species. AB - 1. The effect of 70 mg/l and 35 mg/l MS 222 an anaesthetic on the enzymes: superoxide dismutase (SOD), catalase (C) and peroxidase (P) were estimated in erythrocytes of Cyprinus carpo, a freshwater fish and Dicentrarchus labrax, a marine fish. 2. The end of the summer, at 16 degrees C MS 222 in concentration 70 mg/l caused an enhancement of the SOD and peroxidase activities and a decrease of the catalase activity. 3. In the autumn at 22 degrees C SOD and peroxidase activities in erythrocytes of Dicentrarchus labrax are normally higher than at 16 degrees C. On the contrary MS 222 causes no significant modification of enzymatic activities measured, but an increase in the dispersion of the results. 4. At 13 degrees C in the spring, MS 222 has no immediate influence on the activity of these enzymes, whilst at the same temperature at the beginning of winter, SOD is the only one activated. 5. It seems that in experiments concerning peroxide metabolism enzymes the use of anaesthetic MS 222 is not advisable. PMID- 2566444 TI - Effect of bleached kraft pulp mill effluent on hepatic biotransformation reactions in vendace (Coregonus albula L.). AB - 1. The effect of bleached kraft pulp mill effluent (BKME) on xenobiotic biotransformation enzyme activities in the liver of vendace (Coregonus albula L.) was studied by exposing some fish in field laboratory tanks at 0, 0.1, 0.2 and 0.5% (v/v) effluent concentrations of BKME corresponding to 0-0.08 toxic units and others in clean water as controls. 2. Slight increase (57%, 68%) in polysubstrate monooxygenase enzyme activities, measured as benzo(a)pyrene hydroxylase and 7-ethoxycoumarin O-deethylase activities, was observed in a dose related manner after 70 days of exposure to 0.5% concentration of BKME. 3. Highest increase was observed at 0.2% effluent concentration after 120 days of exposure, measured as benzo(a)pyrene hydroxylase activity. 4. BKME had an affect on UDPglucuronosyltransferase, at the beginning (14-70 days) by decreasing and in longer exposure (120 days) by slightly increasing the activity. PMID- 2566445 TI - Linearity of the accumulation of various dosages of uranium in the major organs of mature male Japanese quail. Effect of various doses of estradiol-17 beta. AB - 1. Male Japanese quail were given 2.20 x 10(-4)-14.53 mg uranium/kg intravenously as uranyl ion (235U label). 2. The relationship between dosage and the 18-hr accumulation of U in leg bones, liver, kidneys and testes was linear. 3. Increases in U deposition with increased dosage were approximately 1:1, except for kidneys where 10-fold increases in dosage resulted in 25-fold increases in deposition. 4. Estradiol-17 beta increased U deposition in bones by 15-fold thereby providing some protection for the kidneys as now the ratio of dosage to accumulation was 1:1. PMID- 2566446 TI - Twitch potentiation of frog (Rana japonica) skeletal muscle by antipyrine. AB - 1. The effects of antipyrine on single muscle fibres from the frog Rana japonica were examined. Antipyrine potentiated the twitch tension, but did not affect the tetanus response or induce any contracture by itself. 2. The duration of the action potential was prolonged in a dose-dependent manner. 3. Antipyrine did not affect the mechanical threshold, the uptake of Ca2+, or the Ca2+ induced release of Ca2+ from the sarcoplasmic reticulum. 4. Ca2+-channel blockers did not affect the twitch potentiation by antipyrine. PMID- 2566447 TI - Characterization of the normal bovine platelet aggregation response. AB - 1. Bovine platelets are more sensitive to stimulation by platelet activating factor (PAF) than adenosine-di-phosphate (ADP) or thrombin. 2. While epinephrine, arachidonic acid and serotonin are ineffective by themselves as aggregatory stimulants of bovine platelets they enhance the aggregation response of other platelet agonists. 3. There is no correlation between thromboxane A2 production and release and the extent of platelet aggregation in bovine platelets. 4. The dependence of bovine platelet aggregation on a phospholipid pathway and calcium mobilization is indicated. PMID- 2566448 TI - Temperature regulation following nickel intoxication in the mouse: effect of ambient temperature. AB - 1. The purpose of this study was to examine the interaction between ambient temperature (Ta) and the effects of nickel chloride on the thermoregulatory system of the mouse. 2. Male mice of the BALB/c strain were injected with nickel chloride at dosages of 0, 0.1, 1.0, 2.5, 5.0 and 10.0 mg/kg intraperitoneally and placed in an environmental chamber set at a Ta of either 10, 20, 30 or 35 degrees C for 60 min. Colonic temperature was then measured after one hour of exposure at a given Ta. 3. The thermoregulatory effects of nickel chloride were highly dependent on Ta. Nickel chloride had no effect on body temperature at Ta's of 30 and 35 degrees C. 4. 10 mg/kg dosage of nickel chloride caused a significant reduction in colonic temperature at a Ta of 20 degrees C. At a Ta of 10 degrees C the 5 and 10 mg/kg dosages of nickel chloride caused a significant lowering of body temperature. 5. Using segmented linear regression techniques it was shown that the threshold dose of nickel chloride for causing hypothermia was 9.6 and 3.3 mg/kg at Ta's of 20 and 10 degrees C, respectively. 6. This study has shown that two stressors, low Ta and nickel chloride intoxication, when applied independently have no effect on body temperature; however, when applied simultaneously, they have a significant toxic effect on thermoregulation. PMID- 2566449 TI - A vinblastine sensitive high affinity choline uptake system. AB - 1. The Limulus cardiac ganglion high affinity choline uptake system (HAChUS) was inhibited 40, 51 and 64% following pre-exposure to 10, 100 and 500 microM vinblastine, respectively. 2. In contrast, high affinity uptake of choline in the Limulus corpora pedunculata and abdominal ganglia, tissues in which a cholinergic function has been described, were unaffected. 3. In pulse-chase experiments, the cardiac ganglion was incubated in 0.1 microM [3H]choline for 60 min and then switched to an incubation medium containing 1 mM unlabelled choline for varying periods of time. 4. Under these conditions, a 3-fold increase of radiolabel above basal level was measured in the pellet fraction within 2 hr of post-labelling incubation. 5. Prior exposure of the ganglion to 500 microM vinblastine completely eliminated this increase of radioactivity in the pellet fraction. 6. Treatment of the radiolabelled pellet fraction with phospholipase C resulted in the solubilization of 72% of the radiolabel. 7. Ten (10) microM 5 hydroxytryptamine (5-HT), a concentration previously shown to inhibit spontaneous electrical activity within the cardiac ganglion, resulted in a 40% decrease in high affinity choline uptake in this tissue selectively. 8. These results are consistent with the view that a probable role of the Limulus cardiac ganglion HAChUS is the supply of choline subserving the synthesis of membrane phospholipid. 9. It is further speculated that this membrane phospholipid synthesis may be associated with synaptic vesicle turnover. PMID- 2566450 TI - Testosterone as a factor modifying the toxicity of ekatin for the Pharaoh quail (Coturnix coturnix Pharaoh). I. Mature birds. AB - 1. Testosterone reduces the haemolytic action of Ekatin on the morphotic blood elements by accelerating maturation of erythroblastic cells. 2. Testosterone, by accelerating the metabolism of the pesticide causes a defective defence processes of the organism by decreasing the number of macrophages. 3. Intoxication of androgenised birds with Ekatin leads to a breakdown of the systemic adaptive mechanisms by rapid and early switching on the adrenal cortex in response to the stress caused by poisoning. PMID- 2566451 TI - Further characterisation of the [35S]-TBPS binding site of the GABA receptor complex in locust (Schistocerca gregaria) ganglia membranes. AB - 1. Using homogenates of supraoesophageal ganglia from locust we observed specific binding of [35S]-TBPS which was linear with protein concentration up to 7 mg/ml, showed a pH optimum at pH 9.0 and was linear with NaCl concentration up to 350 mM. 2. Kinetic analysis of the binding showed positive cooperativity as a result of changes in on and off-rates with occupation of the binding site by the ligand. The apparent KD = 417 nM and Bmax = 1083 fmol/mg of membrane protein were calculated using a computer program for dose-response curve fitting. 3. The binding was enhanced by GABA, pentobarbital and benzodiazepines. Picrotoxinin had no effect on the binding at 0.1 mM. Only the cage convulsants TBPS and IBP were able to displace the binding. 4. Whilst the characteristics of the binding are similar to those reported for house fly thorax and abdomen preparations they are significantly different from those reported for house fly head, cockroach nerve cord and rat brain. PMID- 2566452 TI - Vasoactive properties of dihomo-gamma-linolenic acid and series one prostaglandins in a freshwater teleost, Channa maculata. AB - 1. Prostaglandins A1, B1, E1 and F1 alpha (2-120 micrograms/kg), arachidonic acid and dihomo-gamma-linolenic acid (0.1-2 mg/kg) were injected intravenously into Channa maculata and changes in arterial blood pressure were recorded. 2. Injection of PGF1 alpha had no significant effect on arterial blood pressure. Injection of PGA1 and PGE1 was followed by dose-dependent hypotension whereas injection of PGB1 elicited significant dose-dependent increase in arterial blood pressure. 3. Both dihomo-gamma-linolenic acid and arachidonic acid were also depressor agents but dihomo-gamma-linolenic acid was more potent. 4. A single bolus intravenous injection of indomethacin (5 mg/kg) or 4 daily intraperitoneal injections (4 x 10 mg/kg) significantly lowered arterial blood pressure. One hour after pre-treatment of indomethacin, the vascular effects of both prostaglandin precursors were abolished. 5. It appears that the vascular effects of prostaglandins in Channa maculata are qualitatively different from those reported for mammals. PMID- 2566453 TI - Computer analysis of the effect of beta-endorphin and dynorphin and related compounds on opioid binding to mouse brain membrane. AB - The binding of three tritiated opioid agonists--dihydromorphine, D-ala2-D-leu5 enkephalin and ethylketocyclazocine--was subjected to competition by unlabeled beta-endorphin, dynorphin-(1-13), and various fragments of these peptides, and the results analyzed by a computer program that we developed in an earlier study [11]. Peptides in both groups bound with highest affinity to sites 1 and 3 in our 4-site model, corresponding to the mu and delta sites of conventional classifications, with the dynorphin peptides also interacting with site 2, the kappa site. These results are discussed in relationship to the possible biological roles of these peptides as analgesics or as modulators of analgesia. PMID- 2566454 TI - Approximate maximum likelihood population pharmacokinetic models for the design of dosage regimens. AB - One of the applications of pharmacokinetics and pharmacodynamics is the design of improved drug administration regimens. In its simplest form only global descriptors of the response to a bolus are used to obtain the dosing scheme leading to the desired drug plasma concentration level. For further improvement on these simple administration regimens it is necessary to have additional information on the drug kinetics in terms of either a disposition equation model or a compartmental model. Due to the high variability of individual responses to the same dosing scheme the design of drug administration regimens is often based on the average pharmacokinetic parameters of a data base of individuals. Although widely used, this approach presents some problems since the average disposition equation model and the average compartmental model are inconsistent between themselves and with the global descriptors of the average response. In this paper methods and algorithms for the estimation of alternative population pharmacokinetic models are developed and their use is illustrated using pharmacokinetic data available from the literature. PMID- 2566455 TI - [Subtotal adrenalectomy versus autotransplantation of the adrenal cortex--an alternative procedure in bilateral adrenalectomy in MEN II?]. AB - Complete loss of steroid production is a very remarkable effect of total adrenalectomy. This is the reason why attempt was made to preserve some adrenal cortex function in cases with bilateral adrenalectomy. Till now successful autografting of adrenal cortex has been reported only in cases with Cushing's syndrome. We now report on two cases with MEN II syndrome, in whom autografting into the rectus muscle was done successfully. In order to compare results, data on two patients with subtotal adrenalectomy are given. Outcome was similar. In all 4 cases function of adrenal cortex was demonstrated by scintigraphy and by blood chemistry. In one case with autotransplantation and in another with subtotal resection, the remaining adrenal tissue showed response to ACTH. Provided more experience, in our opinion subtotal adrenalectomy and adrenal autotransplantation in cases of MEN II syndrome might turn out to prove as equivalent procedures. PMID- 2566458 TI - Displacement of bilirubin from adult and newborn serum albumin by a drug and fatty acid. AB - Competitive binding of bilirubin and 5-butyl-1-cyclohexylbarbituric acid (bucolome) to human serum albumin was investigated by equilibrium dialysis, with and without added bilirubin (0.5 mol/mol albumin). Further observations were made by peroxidase oxidation kinetics. Finally, cobinding of the two ligands was studied by spectrophotometry. Experiments were performed with defatted adult human serum albumin, with umbilical cord serum, and with adult albumin to which was bound 2 mol palmitate/mol albumin. Bucolome does not induce displacement of bilirubin from binding to defatted adult albumin when one molecule of either ligand is bound. The second molecule of bound bucolome displaces the first bilirubin molecule. With albumin in umbilical cord serum, the first bucolome does decrease binding of the first bilirubin to some extent although the two ligands can bind to the same albumin molecule. The presence of two molecules of bound palmitate together with one of bilirubin causes enhanced bilirubin displacement on binding of bucolome. These observations may be of consequence for practical testing of the bilirubin displacing effect of drugs. Experiments with neonatal albumin and with added fatty acid should be included for a complete study. PMID- 2566457 TI - Characterization of in vivo acid secretory responses of rabbit with comparison to dog and rat. AB - Even though rabbit gastric glands are a commonly used model for the study of gastric physiology, little is known about the secretion of gastric acid in the rabbit in vivo. Gastric acid secretion in response to pentagastrin, histamine, and bethanechol stimulation, and the effect of specific cholinergic and histamine H2-receptor blockers was studied in 62 urethane-anesthetized rabbits. Additionally, the responses of eight conscious rabbits prepared with chronic gastric fistula were compared with similarly prepared conscious dogs (n = 10) and conscious rats (n = 18). In anesthetized rabbits, maximal pentagastrin stimulation resulted in acid outputs only 22% that of maximal histamine stimulation, but this was enhanced by restoring cholinergic tone with a subthreshold infusion of bethanechol. In conscious rabbits, pentagastrin was an effective stimulant, resulting in maximal acid output 70% that of maximal histamine stimulation. This is in contrast to the in vitro findings reported reported by others utilizing isolated gastric glands and cells. Histamine was a potent stimulant of acid secretion in both the anesthetized and conscious rabbit, an observation that parallels the in vitro findings. Unlike the dog and rat, atropine was ineffective as an inhibitor of histamine-stimulated acid secretion in the conscious rabbit, although it was marginally effective against histamine in anesthetized rabbits and against pentagastrin in conscious rabbits. It is concluded that cholinergic tone plays a crucial role in pentagastrin-stimulated acid secretion in the rabbit. This may be an explanation for the poor response to pentagastrin described in isolated rabbit gastric gland preparations. PMID- 2566456 TI - Relaxatory responses of canine proximal stomach to esophageal and duodenal distension. Importance of vagal pathways. AB - The viscerovisceral reflex control of gastric tone remains poorly characterized. We have previously demonstrated physiological variations in gastric tone that occur during fasting and after feeding. These variations are neurally regulated. We have now compared the reflex mechanisms modulating gastric tone that are elicited by esophageal or duodenal distension in fasted, conscious dogs. To determine the pathways involved in these reflexes, we combined the technique of vagal blockade (by cooling the supradiaphragmatic vagi isolated within a surgically implanted cooling jacket) with the administration of autonomic drugs. Gastric tone was measured as the air volume within an intragastric bag maintained at a constant, low pressure by an electronic barostat. Standardized distensions were performed by means of an inflatable balloon-catheter positioned either in the mid-esophagus (in three dogs) or in the distal duodenum (in three dogs). A profound and consistent gastric relaxation was induced by distension of either the esophagus (247 +/- 21 ml delta volume, P less than 0.05) or the duodenum (238 +/- 29 ml, P less than 0.05). Supradiaphragmatic vagal cooling abolished the gastric relaxatory response to duodenal distension and significantly reduced, but did not completely suppress, the response to esophageal distension. Neither cholinergic stimulation (intravenous bethanechol) nor adrenergic blockade (combined intravenous phentolamine and propranolol) had any significant effect on either gastric relaxatory response. Combined adrenergic and cholinergic (intravenous atropine) blockade induced gastric relaxation, but failed to suppress the gastric responses. We conclude that both esophageal and duodenal distension elicit gastric relaxation by a noncholinergic vagal mechanism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566459 TI - Stimulative effects of TMB-8 and trifluoperazine on pancreatic hormone release. AB - The effects of 8-N-N-diethylamino octyl 3,4,5-trimethoxybenzoate (TMB-8) and trifluoperazine (TFP) on the early phase (10 min) of the release of pancreatic hormones from isolated rat islets were investigated. TMB-8 and TFP stimulated insulin, glucagon, and somatostatin release in a dose-dependent manner at a low glucose concentration (2.5 mM). The levels of glucagon and somatostatin release were also stimulated by these two agents at a high glucose concentration (10 mM). Their effects were independent of external calcium ion level. These two agents did not modify insulin release at the high glucose concentration. The stimulative effects of the two agents on the release of these hormones were partially suppressed when the islets were pretreated with 6-hydroxydopamine (6-OHDA), a chemical adrenergic denervator that acts at nerve endings. In this situation, the norepinephrine (NE) released from pancreatic islets decreased to 44% of that of non-treated islets (P less than 0.01). The addition of NE (10(-9) M) to the incubation medium increased insulin, glucagon, and somatostatin secretion by 20 30% over control levels (P less than 0.05). In conclusion, the early phase of pancreatic hormone release was stimulated by TMB-8 and TFP. Our results strongly suggest that these two drugs could be mediated by the NE released from nerve endings in the islets. PMID- 2566460 TI - Effect of tick-borne fever (Ehrlichia phagocytophila) and trypanosomiasis (Trypanosoma brucei 1066) on the pharmacokinetics of sulfadimidine and its metabolites in goats. AB - The effect of tick-borne fever (TBF) and trypanosomiasis (TBR) on the plasma disposition of sulfadimidine (SDD) in goats was studied after iv administration of 20 and 200 mg/kg of body weight. In each group of six goats, the plasma disappearance curves showed four animals with rapid and two with slow SDD elimination. It is likely that this difference is determined by oxidative rather than acetylation phenotype. In all goats administered 20 mg/kg, half-life increased with TBF but not with TBR. Vd(beta) decreased with both infections. With 200 mg/kg, Vd did not change, whereas AUC and MRT increased with both infections. Metabolites were examined in TBF experiments: N4-acetyl-SDD (N4Ac), 6 hydroxymethyl-SDD (CH2OH) and its glucuronide, 5-hydroxy-SDD (SOH) and its glucuronide, and 6-carboxy-SDD (COOH) and its glucuronide (COOH-glu). At low dose (20 mg/kg), TBF caused the proportion of dose recovered from urine as unchanged SDD to be halved, whereas N4Ac increased correspondingly (2x). After the high dose (200 mg/kg), elimination was saturated and changes in proportional recovery of SDD in urine were less. However, the N4Ac proportion was still doubled, in contrast to the other metabolites, suggesting that saturation was caused by oxidations rather than by acetylation. Formation of CH2OH was the same in health and disease after the low dose, but glucuronidation dropped from 20% to 4% (rapid) or 7% (slow) of total CH2OH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566461 TI - Pulmonary toxicity of trichloroethylene in mice. Covalent binding and morphological manifestations. AB - We examined the time course of trichloroethylene (TCE)-induced pulmonary injury and focused on morphological changes and covalent binding of [14C]TCE soon after administration of a single dose of TCE (2000 mg/kg) to CD-1 male mice. At 1 hr after chemical treatment, Clara cells of the bronchiolar epithelium exhibited necrotic changes involving the mitochondria and endoplasmic reticulum. Dilatation of the endoplasmic reticulum became more severe at 2 hr after TCE administration and, by 4 hr, distended cisternae coalesced to form small vacuoles within the cytoplasmic matrix of the Clara cell. The severity of cellular damage increased progressively between 8 and 12 hr and, by 24 hr, the majority of Clara cells within an airway were severely vacuolated. Covalent binding of [14C]TCE to lung macromolecules was evident at 1 hr, peaked at 4 hr, declined thereafter, and reached a plateau between 12 and 24 hr. Peak binding (142.6 +/- 31.8 nmol/g of wet weight) represented approximately 20% of [14C]TCE distributed to the lung. Although the levels of binding in the liver were at all times greater than those in the lung, liver injury was relatively insignificant. The results demonstrate a positive correlation between the onset of Clara cell injury and the formation of reactive metabolites, as assessed by covalent binding of [14C]TCE. PMID- 2566462 TI - Hepatic drug metabolism in female Fischer rats as a function of age. PMID- 2566463 TI - Metabolism of atrial natriuretic peptide. Extraction by organs in the rat. AB - The pharmacokinetics and metabolism of atrial natriuretic peptide (ANP24) were investigated in male Sprague-Dawley rats. Animals were instrumented with arterial and venous catheters to infuse ANP24 and sample blood at various systemic locations; ANPir concentrations were determined by radioimmunoassay. Total clearances (TC) were 150 +/- 15 and 90 +/- 3 ml of blood/min/kg at infusion rates of 137 and 833 ng/min/kg and the half-life of ANPir was 35 +/- 5 seconds. The volume of distribution was 87 +/- 8 ml/kg at the high infusion rate. The kidneys' extraction ratios (E) of ANPir at the two rates of infusion were 0.55 and 0.61, respectively, whereas the intestines' were 0.44 and 0.27. E values of the muscle/sex organs were 0.43 and 0.54 at the two infusion rates. In contrast, no significant degree of extraction was observed for the liver or the heart and lungs. Taking regional blood flow into account, the kidneys were responsible for one sixth to one third of TC while the intestines accounted for one sixth of TC. We conclude that ANP24 is cleared from the blood at a relatively high rate under steady state conditions and that the kidneys, intestines, and muscle/sex organs contribute to its metabolic fate. PMID- 2566464 TI - Activities of cytosolic and microsomal drug oxidases of rat hepatocytes in primary culture. AB - Sensitive and specific chromatographic assays for the measurement of flavin containing monooxygenase (N-oxygenase and S-oxygenase activities) and aldehyde oxidase activities in rat hepatocyte primary cultures were developed. Conditions for the measurement of enzymatic activities in rat liver cell cultures were first optimized using freshly isolated cell suspensions. Activities of the cytochrome P 450/P-448 isozymes in rat hepatocytes maintained in primary culture [assessed by the O-deethylation of 7-ethoxycoumarin (P-450/P-448) and the N-demethylation of N,N-dimethylaniline (P-450)] rapidly declined to 25% of the initial levels by 48 hr in culture. The flavin-containing monooxygenase system was considerably more stable in cell culture. Flavin N-oxygenase activity (assessed by the N-oxidation of N,N-dimethylaniline) declined slightly (10-15%) and remained almost constant over the 48-hr culture period, whereas S-oxygenase activity (assessed by the S oxygenation of tetrahydrothiophen) gradually declined and stabilized at approximately 65% of its initial activity at 48 hr in culture. Aldehyde oxidase activity (assessed by the 1-hydroxylation of phthalazine) declined to approximately 20% of the initial value by 48 hr in culture. The differential stability of the microsomal and cytosolic drug oxidases in rat hepatocytes in primary culture demonstrates some of the limitations of this model for metabolic studies. PMID- 2566465 TI - Selective inactivation of rat liver cytochromes P-450 by 21-chlorinated steroids. AB - The inactivation by 21-chlorinated steroids of rat liver cytochromes P-450 involved in the hydroxylation of progesterone and androstenedione has been investigated. Preincubation of intact liver microsomes from phenobarbital-treated rats with 21-chloropregnenolone, 21,21-dichloropregnenolone, or 21,21 dichloroprogesterone in the presence of NADPH caused a time-dependent decrease in progesterone 21-hydroxylase and in progesterone or androstenedione 6 beta hydroxylase activity but had negligible or only minor effects on five other steroid hydroxylases. The compounds differed, however, with regard to the relative rate constants for inactivation of the 21- and 6 beta-hydroxylases. For example, 21,21-dichloroprogesterone and 21,21-dichloropregnenolone inactivated the progesterone 6 beta-hydroxylase at similar rates, but the dichloroprogesterone was a more effective inactivator of the 21-hydroxylase. The results indicate that the introduction of a dichloromethyl group into a substrate bearing a methyl group normally hydroxylated by only one or a few isozymes of cytochrome P-450 may be a rational means of designing isozyme-selective inhibitors but that target and nontarget enzymes may not totally retain the regioselectivity they exhibit towards the underivatized substrate. PMID- 2566466 TI - Effect of organ perfusion on renal drug transport. Application to furosemide in the isolated perfused rat kidney. AB - Angiotensin II was used as a probe to study the effect of changes in perfusate flow rate on the renal clearance parameters of furosemide in the isolated perfused rat kidney. Drug studies were performed in three rats with no angiotensin II present in the perfusate (treatment I) and in three rats with a 2.7 ng/min infusion of angiotensin II into the perfusate (treatment II). Furosemide was introduced into the recirculating perfusate at an initial concentration of 3.5 micrograms/ml and was assayed using HPLC. The protein binding of furosemide in perfusate was determined by equilibrium dialysis. Angiotensin II was found to have a dramatic effect on the renal hemodynamics, resulting in a 42% decrease in perfusate flow, a 27% decrease in GFR, and a 25% increase in filtration fraction. Values for the fractional excretion of glucose were very low and consistent, with or without angiotensin II (3.0-3.5%). Although the fraction unbound of furosemide was unchanged between treatments (0.770 for treatment I vs. 0.695% for treatment II), the renal and secretion clearances of furosemide were reduced by about 30% in the presence of angiotensin II. However, if the renal clearance (CLr) was corrected for free fraction (fu) and glomerular filtration rate (GFR) [ER = CLr/(fu.GFR)], there was no difference between the excretion ratio (ER) values of furosemide after the two treatments (29.0 for treatment I vs. 29.6 for treatment II). These results imply that the altered clearance parameters of furosemide are more likely the consequence of a reduction in functional nephron mass rather than a change in intrinsic secretory transport per unit mass of nephron. PMID- 2566467 TI - Inhibition of 3-methylindole bioactivation by the cytochrome P-450 suicide substrates 1-aminobenzotriazole and alpha-methylbenzylaminobenzotriazole. AB - The cytochrome P-450 suicide substrates 1-aminobenzotriazole (ABT) and alpha methylbenzylaminobenzotriazole (alpha MB) were used as probes to examine the participation of cytochrome P-450 monooxygenases in the metabolism and covalent binding of 3-methylindole. ABT was a potent inactivator of 3-methylindole turnover and covalent binding of [methyl-14C]3-methylindole to protein in goat lung microsomal incubations. Both covalent binding and 3-methylindole turnover were decreased approximately 50% at 0.01 mM and 100% at 0.1 mM concentrations of ABT. The effects of ABT indicated that toxicity, as related to covalent binding, was directly dependent upon cytochrome P-450 catalysis. The inactivation of 3 methylindole turnover was greater with a 0.01 mM concentration of the isozyme selective inhibitor alpha MB, 74% as compared with 47% for ABT. alpha MB (0.01 mM) decreased benzphetamine N-demethylase activity by 82% but decreased 7 ethoxyresorufin O-deethylase activity by only 28%. Thus, both 3-methylindole metabolism and benzphetamine oxidation were selectively inactivated by alpha MB. These findings suggest that 3-methylindole is metabolized to alkylating, electrophilic intermediates preferentially by the homologues of "phenobarbital inducible" isozymes (presumably forms 2 and 5 in analogy to rabbit lung isozymes) to cytochrome P-450 in pulmonary microsomes, rather than by the polycyclic aromatic hydrocarbon-inducible isozymes. PMID- 2566468 TI - Isolation and identification of 6-desmethylnaproxen sulfate as a new metabolite of naproxen in human plasma. AB - A new metabolite of naproxen, 6-desmethylnaproxen sulfate (6-DMNS), has been identified in plasma from normal and uremic subjects after oral administration of a single 500 mg dose of naproxen. Tentative identification was achieved by the finding of an increase in the concentration of 6-DMN upon incubation of the plasma with arylsulfatase from Helix pomatia or from Aerobacter aerogenes. More definitive identification was established through demonstration that the HPLC retention time of the conjugate is identical to that of an authentic reference sample of 6-DMNS. Unequivocal identification was accomplished by means of LC-MS after the metabolite was isolated from the plasma by protein precipitation with acetonitrile and further purified by anion-exchange and reversed phase HPLC. Plasma profiles of 6-DMNS for normal and uremic subjects, obtained by a procedure involving differential enzymatic hydrolysis using arylsulfatase from H. pomatia, revealed that 6-DMNS was present in plasma from all subjects but in relatively high concentrations only in subjects with impaired renal function and that the extent of the conjugation is related directly to the severity of the dysfunction. No evidence was found for the presence of glucuronide or sulfate conjugates of naproxen in plasma. PMID- 2566469 TI - Fluconazole is a potent inhibitor of antipyrine metabolism in vivo in mice. AB - Fluconazole, a bis-triazole antifungal, is distinguished from imidazole antifungals (e.g. ketoconazole) by its potency and pharmacokinetic characteristics. Imidazole-containing compounds are well documented to inhibit the hepatic cytochrome P-450-dependent enzyme system; whether this effect occurs with a bis-triazole agent is unknown. The [14C]antipyrine breath test was employed to investigate the effects of fluconazole on this enzyme system in CD-1 male mice. Control, ketoconazole (100 mg/kg), and fluconazole (1 and 10 mg/kg) were studied in single- and multiple-dose experiments. Fluconazole had potent inhibitory effects on the total (mean = -73% +/- 2%), demethylase (mean = -90% +/ 2%), and nondemethylase (mean = -60% +/- 4%) elimination rate constants (all p less than 0.001). The fraction of the administered radioactivity excreted as 14CO2 was decreased by 50-80% in the fluconazole groups (p less than 0.001). These effects were seen after single- and multiple-dose studies; however, return to baseline occurred more quickly in the multiple-dose group. These effects were significantly more pronounced than those observed with equipotent doses of ketoconazole. These results provide evidence that fluconazole is a potent, partially selective, and reversible inhibitor of the cytochrome P-450-dependent enzyme system in mice. Future studies will be required to assess this property and possible interactions with drugs metabolized by this enzyme system in humans. PMID- 2566470 TI - Cilastatin-insulin interaction in the rat. AB - The administration of the selective renal dehydropeptidase I inhibitor cilastatin on insulin pharmacokinetics and pharmacodynamics was studied in the intact rat model. After either a single iv dose of cilastatin, 25 mg/kg, or saline, a 30-min urine sample and pre- and postcollection blood samples were obtained from rats infused to steady state with either saline or insulin. There were no significant changes in insulin or glucose handling in groups administered saline with or without cilastatin. In rats with elevated serum insulin concentrations (greater than 75 microU/ml), cilastatin produced a 2-fold decrease in fractional urinary clearance of insulin (1.03 +/- 0.49 vs. 2.30 +/- 0.72%; p = 0.0026). No differences in renal function as assessed by creatinine clearance and sodium transport were observed between the groups. Serum glucose levels decreased by 46% and were significantly correlated with serum insulin levels (r = -0.66; p = 0.003). The administration of cilastatin to a patient exhibiting hyperinsulinemia may decrease the total body clearance of insulin resulting in profound hypoglycemia. PMID- 2566471 TI - Characterization of a carbamic acid ester glucuronide of the secondary amine sertraline. AB - In the dog, the major excretory metabolite of the antidepressant sertraline was characterized as sertraline carbamoyl-O-glucuronide. The intact conjugate was isolated from bile by HPLC. The metabolite was labile to beta-glucuronidase and produced sertraline as the single hydrolytic product, based on HPLC and GC-MS analyses. By fast atom bombardment MS analysis, [M+H]+ and [M+Na]+ ions at m/z 526 and 548 were observed, as were the proton and sodium adducts of the aglycone (m/z 350 and 372) due to cleavage of the glycosidic bond and elimination of the glucuronic acid moiety (176 amu). The observed mass of the aglycone was 44 amu greater than sertraline, indicating that a carbamic acid of this secondary amine was conjugated with glucuronic acid. These data suggest that sertraline in solution reversibly associates with CO2 before formation of sertraline carbamoyl O-glucuronide. This novel amine glucuronide was also identified in human plasma after the oral administration of sertraline to each of seven subjects. The glucuronide was stable in plasma at both acidic and basic pH. PMID- 2566472 TI - Stereoselective oxidation of nilvadipine, a new dihydropyridine calcium antagonist, in rat and dog liver. AB - The stereoselective oxidation of nilvadipine (NV), a new 1,4-dihydropyridine calcium antagonist, to the corresponding pyridine analog was studied after incubation of (+)- and (-)-NV with rat and dog liver microsomes. The rates of formation of the pyridine analog and disappearance of NV were similar for each species, indicating that aromatization of NV is the primary metabolic step. Formation of the corresponding pyridine required the presence of an NADPH generating system and was significantly inhibited by carbon monoxide and metyrapone, indicating the participation of cytochrome P-450. In male rat liver microsomes, the apparent Km values for the formation of the pyridine from (+)- and (-)-NV were 11.2 and 8.1 microM, and the Vmax values were 7.48 and 3.37 nmol/mg of protein/min, respectively. Therefore, the Vmax/Km value, which is equivalent to the intrinsic clearance of the drug, for the oxidation of (+)-NV was 1.59-fold greater than that for the oxidation of the (-)-enantiomer. In female rats, (-)-NV oxidation exhibited two distinct apparent Km values, whereas the that of the (+)-enantiomer did not. The (+)/(-) ratio of Vmax/Km was 1.23. On the other hand, in male dog microsomes the Km values for (+)- and (-)-NV were 21.9 and 12.2 microM, and Vmax values were 3.02 and 2.45 nmol/mg of protein/min, respectively; the (+)/(-) ratio of Vmax/Km was 0.69. These results indicate that the stereo-selective oxidation of NV is species dependent and is sex related in rat liver. PMID- 2566473 TI - Identification of two main urinary metabolites of [14C]omeprazole in humans. AB - The excretion and metabolism of [14C]omeprazole given orally as a suspension was studied in 10 healthy male subjects. An average of 79% of the dose was recovered in the urine in 96 hr, with most of the radioactivity (76% of dose) being eliminated in the first 24 hr. Pooled urine (0-2 hr) from five subjects, containing about 47% of the dose, was analyzed by reverse phase gradient elution LC with radioisotope detection. Omeprazole was completely metabolized to at least six metabolites. The two major metabolites were extensively purified by LC and their structures were determined by MS with derivatization and use of stable isotopes, 1H NMR, and comparison with synthetic references. They were formed by hydroxylation of a methyl group in the pyridine ring, followed by further oxidation of the alcohol to the corresponding carboxylic acid. Both metabolites retained the sulfoxide group of omeprazole, rendering them as unstable as the parent compound at pH less than 7. They accounted for approximately 28% (hydroxyomeprazole) and 23% (omeprazole acid) of the amount excreted in the 0-2 hr collection interval. Based on in vitro studies with the synthetic metabolites in isolated gastric glands, it is unlikely that M1 and M2 will contribute to the pharmacological effect of omeprazole in humans. PMID- 2566474 TI - Selective mechanism-based inactivation of the major phenobarbital-inducible P-450 cytochrome from rabbit liver by phencyclidine and its oxidation product, the iminium compound. AB - Phencyclidine, 1-(1-phenylcyclohexyl)piperidine, was found in this study to be a mechanism-based inactivating agent for cytochrome P-450 form 2, the major phenobarbital-inducible cytochrome of rabbit liver microsomes. This process is highly selective for P-450 form 2, both in microsomes and in the reconstituted enzyme system, in that forms 3a, 3b, 4, and 6 are not affected. However, phencyclidine iminium ion, an oxidative metabolite, inactivates both P-450 form 2 and form 3b in a metabolism-dependent manner. Both phencyclidine and its iminium ion give biphasic kinetics of inactivation with similar rate constants, which supports the hypothesis that the iminium ion is an intermediate in the inactivation of P-450 form 2 by the parent compound. The absorption of the oxidized cytochrome and of the ferrous-carbonyl complex in the visible spectrum are both decreased upon inactivation by phencyclidine, indicating modification of the heme moiety. Several modified hemes produced by the action of phencyclidine were isolated by HPLC. PMID- 2566475 TI - Central nervous system kinetics of atenolol and metoprolol in the dog during long term treatment. AB - Beagle dogs with catheters chronically implanted into the lateral cerebral ventricle were used to study the distribution of atenolol and metoprolol between the cerebrospinal fluid (CSF) and blood plasma over a 24-hr period during long term treatment. The concentration of atenolol declined more slowly in CSF than in blood plasma and the CSF/plasma ratio of atenolol (after iv administration for 7 days) increased from 0.08 +/- 0.02 (2 hr after dose) to 0.83 +/- 0.14 (24 hr after dose) (mean +/- SD). Furthermore, the CSF concentration of atenolol, relative to the plasma concentration, increased during repeated drug administration. The CSF/plasma ratio 24 hr after an iv dose was 0.48 +/- 0.12 on day 1 and 0.83 +/- 0.14 on day 7. The CSF concentration of the more lipophilic beta 1-adrenoceptor antagonist metoprolol was almost the same as the concentration of the drug in blood plasma. After 7 days of oral treatment, the CSF/plasma ratio of metoprolol 24 hr after dosing was 0.81 +/- 0.10. The regional CSF concentration of atenolol along the neuraxis was determined in anaesthetized dogs after acute iv administration of the drug. The atenolol concentration in CSF from the lateral cerebral ventricle was similar to that in the cisterna magna but lower than the concentration in CSF sampled from the lumbar region. It is concluded that the CSF concentration of the moderately lipophilic beta 1 adrenoceptor antagonist metoprolol equilibrates with the plasma concentration of the drug more rapidly compared with the hydrophilic drug atenolol. PMID- 2566476 TI - The pharmacokinetics of salicylate in the pregnant Wistar rat. AB - Sodium salicylate, in a single dose of 50 mg/kg, was administered by iv injection to nonpregnant female and 20-day pregnant Wistar rats. Blood samples (for serum) and urine were collected and analyzed for salicylate, gentisic acid, salicyluric acid, and salicyl glucuronides by HPLC. Pregnant rats showed a significant decrease in body weight-normalized total clearance but no change in absolute total clearance of salicylate. On a body weight-adjusted basis there was a slight increase in the apparent volume of salicylate distribution in pregnancy but this increase becomes highly significant if expressed in absolute terms. The biological half-life of salicylate was significantly increased in late pregnancy. Serum protein binding of salicylate is decreased in pregnancy relative to nonpregnant females but in both groups binding shows a concentration dependence. The partial clearances of both salicyluric and gentisic acids were reduced by pregnancy in the rat whereas that of the salicyl glucuronides appeared unchanged. This latter result in intact pregnant animals contrasts with previously reported decreases in glucuronyltransferase activity in isolated liver preparations from pregnant rats. PMID- 2566477 TI - Temporal variations in the pharmacokinetics of isoniazid and N-acetylisoniazid in rats. AB - The temporal variation in the pharmacokinetics of a single dose administration of isoniazid (INH) and its major metabolite, N-acetylisoniazid (AINH), was investigated at 0900 and 2100 in rats synchronized to a 12-hr light-dark cycle. INH and AINH were administered by the intraarterial and ip routes at a dose of 20 mg/kg, in order to determine the hepatic extraction ratio of both compounds. The mean AUC values of INH obtained after the intraarterial and ip injections of the drug were 42.6 and 8.4 micrograms.hr/ml at 0900 and 30.7 and 12.8 micrograms.hr/ml at 2100, respectively. Consequently, there were temporal variations (mean values at 0900 and 2100) in the clearance (491.6 and 665.3 ml/hr/kg), bioavailability (0.20 and 0.42), hepatic extraction ratio of INH (0.80 and 0.58), and apparent hepatic blood flow (9.7 and 18.6 ml/min). Also, the AUC of AINH was higher at 2100 when INH was injected intraarterially. Likewise, AINH had a lower t 1/2 and AUC values and a higher clearance at 2100, but the hepatic extraction ratio of AINH was 0.26 at 2100 compared with the value of 0.07 at 0900. No diurnal variations were present in the apparent volume of distribution of INH and AINH. In vitro experiments showed that the mean activity of the hepatic N-acetyltransferase was 135.3 nmol/mg at 2100, compared with 59.0 nmol/mg at 0900.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566479 TI - [Schizophrenia and selective attention]. AB - A comparative study of selective attention was made on 22 schizophrenic inpatients, 18 non schizophrenic inpatients and a control of 22 non psychiatric adults. Signs of schizophrenia were quantified through the BPRS (Brief Psychiatric Rating Scale) and selective attention was expressed as the capacity to resist a distracting stimulus dimension in the Stroop word-colour test. The results indicated higher distractibility in schizophrenic patients, and a positive, significative relation was found between this interference and the "activation" cluster. PMID- 2566478 TI - In vivo and in vitro pharmacokinetic differences between four structurally closely related anthracyclines in hematopoietic cell subtypes in humans. AB - Minor differences in chemical structure of adriamycin (ADM), 4'-epiadriamycin (E ADM), daunomycin (DNM), and 4-demethoxydaunomycin (D-DNM) lead to large differences between cellular and plasma pharmacokinetic parameters in vivo, as well as in cellular drug handling. Anthracyclines accumulated in cells to several hundred-fold the plasma concentration. Half-lives, as well as the ratio of parent drug/metabolite, differed markedly. The slopes of the in vivo cellular concentration-time curves after the end of the bolus injection resembled the efflux curves observed after a 5-min exposure in vitro. In vivo, the area under the cellular concentration-time curve (AUCc) for equimolar dosages was largest for ADM and smallest for D-DNM. In vitro however, cellular drug levels and AUCc were highest for D-DNM, followed by DNM, E-ADM, and ADM. Final cellular drug concentrations were 300-2500 times the medium concentration, with clearly higher values observed after the 360-min exposure. Minor structural differences were related to considerable variations in cellular drug handling, with different patterns in vivo and in vitro. These studies point to difficulties occurring in the in vitro experimental studies of in vivo pharmacokinetic properties of anthracyclines and stress the need for direct determination of target cell drug concentrations in vivo, in the search for the understanding of cell drug handling related mechanisms of action of the anthracyclines. PMID- 2566480 TI - Bidirectional transhemispheric interaction between amygdaloid kindling induced by excitatory amino acids and electrical stimulation. AB - Using transhemispheric transfer and post-transfer interference as specific measures, the relationship between chemical and electrical kindling at the amygdala (AM) in rats was examined. Chemical kindling was accomplished by repeated injections of a subconvulsive dose of combined L-glutamate (GLU) and L aspartate (ASP) in a molar ratio of 1:3 (GLU 0.375 microM:ASP 1.125 microM). Strong bidirectional transhemispheric transfer between electrical and chemical kindling occurred at the secondary-site AM. In addition, at the primary-site retest, bidirectional posttransfer interference between electrical and chemical kindling was observed. These findings strongly suggest that endogenous GLU/ASP participates in the transsynaptic changes which are presumed to underlie the kindling phenomenon. PMID- 2566482 TI - Assessment of anaesthetic adequacy with upper facial and abdominal wall EMG. AB - We compared changes in biopotentials arising from upper facial (FEMG) and abdominal (AEMG) muscles associated with alterations in alveolar enflurane concentration and neuromuscular block. Induction of anaesthesia significantly reduced both FEMG and AEMG mean amplitudes (-60% and -43%, respectively). Neuromuscular blocker-induced abolition of the electrically evoked thenar EMG response did not prevent FEMG and/or AEMG activation during endotracheal intubation. Decreasing the alveolar enflurane concentration was associated with an increase in FEMG amplitude prior to visible signs of arousal in half of the patients. Movement and other signs of inadequate anaesthesia were associated with distinct increases in FEMG amplitude in 29 out of 30 patients. Recovery from neuromuscular block during unchanged alveolar enflurane concentration was associated with increasing amplitudes of both FEMG and AEMG. Finally, very low amplitude FEMG recordings were always associated with relaxed abdominal muscles. PMID- 2566481 TI - Inducible in vivo DNA footprints define sequences necessary for UV light activation of the parsley chalcone synthase gene. AB - We began characterization of the protein--DNA interactions necessary for UV light induced transcriptional activation of the gene encoding chalcone synthase (CHS), a key plant defense enzyme. Three light dependent in vivo footprints appear on a 90 bp stretch of the CHS promoter with a time course correlated with the onset of CHS transcription. We define a minimal light responsive promoter by functional analysis of truncated CHS promoter fusions with a reporter gene in transient expression experiments in parsley protoplasts. Two of the three footprinted sequence 'boxes' reside within the minimal promoter. Replacement of 10 bp within either of these 'boxes' leads to complete loss of light responsiveness. We conclude that these sequences define the necessary cis elements of the minimal CHS promoter's light responsive element. One of the functionally defined 'boxes' is homologous to an element implicated in regulation of genes involved in photosynthesis. These data represent the first example in a plant defense gene of an induced change in protein--DNA contacts necessary for transcriptional activation. Also, our data argue strongly that divergent light induced biosynthetic pathways share common regulatory units. PMID- 2566483 TI - Serological response to P-fimbriae of Escherichia coli in patients with urinary tract infections. AB - The antibody response to P-fimbriae of Escherichia coli in patients with upper urinary tract infections was investigated. In the sera of patients with pyelonephritis obtained at the initial visit to hospital (3 to 7 days after the onset of symptoms), a high incidence of antibodies to P-fimbriae was detected (12 out of 14 patients). P-fimbriated Escherichia coli strains were isolated from urine samples in all of these antibody-positive patients. Antibodies detected by ELISA using purified antigen were essentially IgG and specifically recognized P fimbriae. These antibodies inhibited completely, or in some cases partially, mannose-resistant hemagglutination with P-fimbriated Escherichia coli. PMID- 2566484 TI - In vitro activity of mannan sulfate, a novel sulfated polysaccharide, against human immunodeficiency virus type 1 and other enveloped viruses. AB - Mannan sulfate, a novel sulfated polysaccharide, was prepared and investigated for its activity against human immunodeficiency virus type 1 (HIV-1) in vitro. Mannan sulfate completely inhibited HIV-1-induced cell destruction and viral antigen expression in HIV-1-infected Molt-4 (clone 8) cells at a concentration of 4 micrograms/ml. The 50% antiviral effective doses obtained with mannan sulfate in Molt-4 (clone 8) cells and in MT-4 cells were 1.5 and 9.3 micrograms/ml, respectively. No toxicity for Molt-4 (clone 8) cells or MT-4 cells was observed at a concentration of 4,000 and 2,500 micrograms/ml, respectively. Mannan sulfate was also inhibitory to other enveloped viruses, i.e. herpes simplex virus types 1 and 2, vaccinia virus and vesicular stomatitis virus. These results suggest that mannan sulfate may be useful for the chemotherapy of various viral infections, including those causing and associated with AIDS. PMID- 2566485 TI - Cancer prevention by mass screening for gynaecologic cancers in the European community. PMID- 2566486 TI - L-alpha-aminodipate antagonizes the priming effect of quisqualate in hippocampal slices. PMID- 2566487 TI - Hyperosmolarity but not histamine evokes secretion of nasal fluid in the rat. AB - Histamine (dihydrochloride or free base), mannitol or sodium chloride applied topically to the nasal mucosa of anaesthetized rats stimulated the secretion of fluid in a dose-dependent manner. Compound 48/80 applied in the same way and histamine given s.c. failed to evoke secretion. The secretion evoked by topically applied histamine was resistant to pretreatment with H1 and H2 receptor antagonists i.p. Furthermore, the secretion evoked by histamine and sodium chloride was resistant to pretreatment with atropine i.p. or lidocaine topically. The secretion evoked by mannitol or by sodium chloride was not reduced by H1 receptor antagonists and was not significantly different from that obtained after challenge with histamine of equivalent osmolality. We conclude that the response to topically applied histamine reflects the hyperosmolarity of the compounds applied and not the effect of histamine on histamine receptors. PMID- 2566488 TI - Biphasic effect of D-2 agonist quinpirole on locomotion and movements. AB - The effects of the D-2 agonist quinpirole on forward progression, and on vertical and lateral movements, were measured for 2 h in rats injected with either saline, 0.03, 0.125, 0.5 or 8 mg/kg of the drug. Results showed that the drug had a biphasic effect: the lowest dose decreased and the high doses increased the amount of locomotion and of movement. The decrease in activity produced by the low dose had its onset within minutes after administration of the drug; persistent and marked hyperactivity was observed from about 60-80 min after injection of 0.5-8 mg/kg of quinpirole. Moreover, in animals injected with intermediate doses, the excitation was preceded by a brief period of reduced locomotion. It is suggested that the biphasic effect may reflect two independent actions of the drug, possibly on activity in the nucleus accumbens. PMID- 2566489 TI - SKF 38393: vacillatory behavior in immature rats. AB - SKF 38393 was administered to postnatal rat pups on day 15 and day 30. Behaviors measured on day 15 revealed that SKF 38393-treated groups displayed significant deficits, in comparison to controls, in spontaneous alternation and decision making: less alternation and more vacillatory behavior at every dosage tested. On day 30, all SKF 38393-related behaviors disappeared. PMID- 2566490 TI - Somatostatin inhibition of VIP-induced somatostatin release, cyclic AMP accumulation and 45Ca2+ uptake in diencephalic cells. AB - The effect of somatostatin (SRIF) on VIP induction of SRIF secretion, cyclic AMP accumulation and 45Ca2+ influx was investigated in cultured diencephalic cells. [D-Trp8]SRIF suppressed VIP-stimulated SRIF release and decreased VIP-stimulated cyclic AMP accumulation in a dose-dependent manner. SRIF-14 blocked basal and VIP stimulated 45Ca2+ entry into cells. The data suggest that the inhibitory effect of SRIF on VIP-induced SRIF release is partly due to a decrease in Ca2+ entry into cells. PMID- 2566491 TI - (+)-[3H]3-(3-hydroxyphenyl)-N-(1-propyl)-piperidine binding to sigma receptors in mouse brain in vivo. AB - Binding of i.v. administered (+)-[3H]3-(3-hydroxyphenyl)-N-(1-propyl)piperidine ([3H]3-PPP) in the brain of intact mice is antagonized dose responsively by sigma receptor ligands. The correlation of potencies for inhibition of binding in vivo and in vitro indicates that sigma receptors in mouse brain are labeled in vivo by i.v. [3H]3-PPP. 3-PPPP, the N-phenylpropyl derivative of norpropyl-3-PPP exhibits very high affinity for sigma receptors in vitro and in vivo. PMID- 2566492 TI - Microdialysis and striatal dopamine release: stereoselective actions of the enantiomers of N-0437. AB - An intracerebral dialysis method was used to test both enantiomers of the very potent and selective dopamine (DA) D-2 agonist 2-(N-propyl-N-2-thienylethylamino) 5-hydroxytetralin, N-0437, for their actions on DA receptors in the striatum of the rat. (-)N-0437 induced a 60% decrease in DA release, which was independent of the presence or absence of a kainic acid lesion placed unilaterally in the striatum. Stereotyped behaviour was apparent following administration of the (-) enantiomer. Thus, (-)N-0437 displayed an agonistic action on both pre- and postsynaptic D-2 receptors. (+)N-0437 did not induce any effect in the release model after peripheral administration nor did it induce any form of stereotypy. A comparison between the effects of (-)N-0437 after oral (10 mumol/kg) and transdermal (10 mumol/kg) administration showed the advantages of the latter mode of administration. Transdermal application induced a much longer duration of action of the drug (13 h) in comparison with the oral mode (5 h). Thus, transdermal administration may be a very useful method of drug application for therapeutic use. PMID- 2566494 TI - Repeated amphetamine: reduced dopamine neuronal responsiveness to apomorphine but not quinpirole. AB - Extracellular recordings from single nigrostriatal dopamine (DA) neurons in rats revealed significantly reduced neuronal sensitivity to the inhibitory effects of i.v. apomorphine following repeated amphetamine (4 mg/kg per day i.p., 14 days). This effect was reversed by acute SCH 23390. Quinpirole sensitivity was reduced in amphetamine-treated rats only following acute SKF 38393 pretreatment. These results suggest that, in amphetamine-treated animals, D-1 receptor activation is important for the expression of reduced nigrostriatal DA neuron sensitivity to apomorphine. PMID- 2566493 TI - NMDA antagonists block restraint-induced increase in extracellular DOPAC in rat nucleus accumbens. AB - The effects of the N-methyl-D-aspartate (NMDA) receptor antagonists CPP, TCP, PK 26124 and ifenprodil, and of the minor tranquillizer diazepam on stress-induced changes of dopamine metabolism in the nucleus accumbens were investigated in the rat. Dopamine metabolism was assessed by measuring the extracellular levels of 3,4-dihydroxyphenylacetic acid (DOPAC) by means of in vivo differential pulse voltammetry with electrochemically pretreated carbon fiber electrodes. Physical immobilization of the rats for 4 min caused a marked and long-lasting increase in extracellular DOPAC levels in the nucleus accumbens. A similar, though shorter lasting, augmentation of extracellular DOPAC was observed in the nucleus accumbens after systemic administration of the anxiogenic agent methyl-beta carboline-3-carboxylate (beta-CCM) (10 mg/kg s.c.). Pretreatment with CPP (1 mg/kg i.p.), TCP (3 mg/kg i.p.), PK 26124 (3 mg/kg i.p.), ifenprodil (3 mg/kg i.p.) or diazepam (2 mg/kg i.p.) totally antagonized the immobilization-induced increase in extracellular DOPAC in the nucleus accumbens. Diazepam and the benzodiazepine (omega 1-2) receptor antagonist flumazenil (30 mg/kg i.p.), but not ifenprodil, also antagonized the beta-CCM-induced activation of dopamine metabolism in the nucleus accumbens. Finally, systemic administration of haloperidol (25 micrograms/kg i.p.) increased the extracellular concentrations of DOPAC in the nucleus accumbens, but pretreatment with ifenprodil (3 mg/kg i.p.) did not modify this response. These data indicate that NMDA receptor antagonists prevent the activation of dopamine metabolism in the nucleus accumbens caused by immobilization stress but not by beta-CCM-induced anxiogenic stimulation. These results suggest that NMDA receptor antagonists may possess an anxiolytic-like action in the rodent, which is exerted via neuroanatomical circuits distinct from those acted upon by diazepam. PMID- 2566495 TI - 5-HT1 receptor agonists attenuate the naloxone-induced jumping behaviour in morphine-dependent mice. AB - 8-Hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) and RU 24969 have been used to investigate whether 5-HT1A and 5-HT1B receptors are involved in the naloxone induced jumping behaviour of the chronically morphine-dependent mouse. To control for possible interactions with catecholaminergic systems, the effects of alpha 1- and alpha 2-adrenoceptor antagonists were investigated. 8-OH-DPAT and RU 24969, as well as buspirone, ipsapirone and flesinoxan, were found to suppress jumping. The effects were mimicked by the alpha-adrenoceptor antagonists, idazoxan, WY 26392, yohimbine and rauwolscine. Inhibition of 5-HT synthesis with para chlorophenylalanine (pCPA) had only minimal effects on withdrawal jumping per se; the attenuating effects of 8-OH-DPAT and RU 24969 were not altered in pCPA pretreated animals. The effects of RU 24969 were blocked by (-)-pindolol and, stereoselectively, by (-)-SDZ 21-009. (-)-Pindolol neither influenced the action of 8-OH-DPAT nor showed any effect per se. The actions of 8-OH-DPAT and buspirone, but not of RU 24969 and idazoxan, were blocked by the 5-HT1A receptor antagonist, spiroperidol. Similarly, both haloperidol and prazosin prevented the attenuating action of 8-OH-DPAT but did not interfere with the action of RU 24969. We conclude that the actions of 8-OH-DPAT and RU 24969 are mediated by postsynaptic receptors. The 5-HT1B receptor appears to mediate the attenuating action of RU 24969; the exact mechanism of action of 8-OH-DPAT remains open but activation of an alpha 1-adrenoceptor is implicated. PMID- 2566496 TI - Involvement of a central nervous pathway in yohimbine-induced insulin secretion. AB - Yohimbine hydrochloride, an alpha 2-adrenoceptor antagonist, was administered (3.3 mg/kg i.v.) to anesthetized normal dogs provided with a T-shaped catheter inserted in the pancreaticoduodenal vein. The effects on blood glucose levels and pancreatic hormones were investigated. We show that yohimbine induced an immediate and pronounced stimulatory effect on insulin secretion accompanied by a clear decrease in blood glucose levels. Yohimbine also stimulated the pancreatic secretion of somatostatin and glucagon. However, the secretion kinetics were not the same for the three hormones: the stimulation was rapid and immediate for insulin and somatostatin, whereas it was progressive for glucagon. All these stimulatory effects were suppressed by propranolol, thus implicating beta adrenergic mechanisms. Bilateral cervical vagotomy markedly reduced the immediate effect of yohimbine on insulin secretion, suggesting that a central neural pathway was implicated. In contrast, the progressive elevation in glucagon secretion was not decreased by vagotomy. Our results suggest that yohimbine stimulates, at least in part, insulin secretion by blocking central alpha 2 adrenoceptors. PMID- 2566497 TI - Alpha 2-adrenoceptor in HT29 human colon adenocarcinoma cell-line: study of [3H]( )-adrenaline binding. AB - The HT29 human adenocarcinoma cell-line was used to investigate the binding of [3H](-)-adrenaline at alpha 2-adrenoceptors. All aspects of the study indicated that alpha 2-adrenoceptors were specifically labeled. [3H](-)-adrenaline bound with high affinity (KD = 2.28 +/- 0.41 nM) to a single population of non interacting sites. The rank order of adrenoceptor antagonists (yohimbine greater than phentolamine much greater than prazosin) and agonists (UK-14,304 greater than clonidine greater than (-)-adrenaline greater than (-)-noradrenaline greater than (+)-adrenaline greater than (+)-noradrenaline greater than amidephrine) to compete with [3H](-)-adrenaline binding showed that the labeled sites were alpha 2-selective and stereospecific. Comparison of the binding parameters of [3H](-) adrenaline with those of [3H]clonidine (partial-agonist) and [3H]yohimbine (antagonist) indicated that [3H](-)-adrenaline and [3H]clonidine labeled a similar number of sites (156 +/- 13 versus 175 +/- 21 fmol/mg protein) and that [3H]yohimbine (Bmax = 246 +/- 22 fmol/mg protein) labeled more sites than the 3H agonists. Data on the inhibition of [3H]yohimbine binding by (-)-adrenaline was better fitted to a two-site model and revealed (1) that the KiL/KiH ratio was higher for (-)-adrenaline than for clonidine (2) that both agonists recognized the same percentage of high-affinity receptors. The results from a kinetic study of [3H](-)-adrenaline binding were apparently inconsistent with the equilibrium data. Both the association and dissociation were bi-exponential, suggesting a relative heterogeneity of the labeled sites. The tritiated physiological full agonist was moreover able to induce tight-binding. The practical consequences of this property are discussed. PMID- 2566498 TI - Effects of adrenalectomy on responses mediated by dopamine D-1 and D-2 receptors. AB - The effects of surgical adrenalectomy were investigated on behavioural responses produced by the selective D-1 agonist, SK&F 38393, alone, and in combination with the D-2 agonist, quinpirole (LY171555). Further, stereotyped responses to apomorphine and LY171555 were assessed following treatment with either the D-1 or the D-2 antagonists, SCH 23390 and raclopride, respectively. There was no difference between sham-adrenalectomized (sham) and adrenalectomized (ADX) groups in responses to SK&F 38393. Although concomitant stimulation of both receptor subtypes increased the incidence of stereotyped sniffing behaviour, there was no difference in the magnitude of this effect between the sham and ADX groups. Raclopride reduced LY171555-induced sniffing and hypothermia less in ADX rats than in sham controls, which was consistent with the hypothesis that adrenocortical hormones affect D-2 receptor responsiveness. SCH 23390 had a greater inhibitory effect on LY171555 responses, but a smaller effect on apomorphine responses in the ADX group compared with their sham controls. It is concluded that the amplified D-2-stimulated response observed in ADX rats may be more dependent on tonic D-1 receptor activation than the control D-2 response of shams. PMID- 2566499 TI - Indirect evidence for a role of prostaglandins as second messengers of the prejunctional effect of opioids in guinea-pig ventricular preparations. AB - The cardiac response to adrenergic nerve stimulation was dose dependently reduced in a statistically significant manner by 1-10 microM dynorphin-(1-13) in isolated atria, and by 0.1-1 microM dynorphin-(1-13) in guinea-pig ventricular preparations. The inhibitory effect of dynorphin was maintained in atria that had been pretreated with two cyclooxygenase inhibitors at concentrations that induce an 80% inhibition of the enzyme, namely indomethacin 3 microM and acetylsalicylic acid 200 microM. The inhibitory effect of dynorphin disappeared in similarly pretreated ventricular preparations. These results suggest that, whilst the mediation of the effect of dynorphin is carried out mainly by specific opioid receptors in the atrial section, in the ventricular tissue it occurs through the endogenous prostanoid system. PMID- 2566500 TI - Time and dose dependence of the 'priming' of the expression of dopamine receptor supersensitivity. AB - The D-1 receptor agonist, SKF 38393 (2 mg/kg s.c.), failed to elicit contralateral turning when administered to drug-naive rats 17 days after unilateral 6-hydroxydopamine (6-OHDA) lesioning of the medial forebrain bundle, while it elicited intense contralateral turning 90 days post-lesioning. On the other hand the D-1/D-2 receptor agonist, apomorphine (0.1 mg/kg s.c.), induced contralateral turning in drug-naive rats lesioned 14 days earlier and made the administration of SKF 38393 (2 mg/kg s.c.) 3 days later effective to evoke contralateral turning ('priming'). The effectiveness of apomorphine as a primer of SKF 38393-induced turning depended critically on the interval between the administration of the two agonists. Effectiveness was minimal after 3 h and increased after 6-12 h, peaked at 72 h and was reduced after 10 days. The D-2 receptor agonist, LY 171555 (0.2 mg/kg s.c.), was also effective as a primer of SKF 38393-induced contralateral turning and this effect also was dependent upon the interval between priming and SKF 38393 administration. Moreover, priming was dependent on the dose of drug used as primer and on the dose of SKF 38393 used as a challenge. In contrast to SKF 38393, priming was unable to make effective a dose of LY 171555 that was ineffective in drug-naive rats, suggesting that LY 171555 affects D-1-dependent turning to a greater extent than D-2-dependent turning. The results indicate that the priming phenomenon is rather strictly time and dose-dependent. PMID- 2566501 TI - Cardiovascular effects of L-tyrosine: influence of blockade of tyrosine metabolism. AB - Tyrosine is the precursor of catecholamines. Small doses of tyrosine produce tachycardia and hypertension while higher doses produce bradycardia and hypotension in anaesthetised rats. The mechanism of these effects has not been established. An increased synthesis and release of catecholamines has been suggested to be the mechanism. Various pretreatments were given to anaesthetised Wistar rats to study the influence of a blockade of L-tyrosine metabolism and thus a blockade of catecholamine synthesis, on these cardiovascular effects: valine, which inhibits tyrosine uptake into brain, alpha-methyl-p-tyrosine, which blocks the rate-limiting enzyme, tyrosine hydroxylase, carbidopa and benserazide, which both inhibit dopa decarboxylase, and desipramine, which blocks catecholamine re-uptake. Benserazide and alpha-methyl-p-tyrosine partially blocked the stimulatory effects of tyrosine. None of the pretreatments were able to block effectively the inhibitory effects of L-tyrosine. Therefore, the metabolism of tyrosine to form catecholamines may be involved in the stimulatory but not in the inhibitory cardiovascular effects of L-tyrosine. Valine pretreatment did not antagonize the depressant effects of tyrosine. Since valine blocks the uptake of L-tyrosine into the brain, the depressant effects of L tyrosine might be peripheral rather than central in origin. PMID- 2566502 TI - Effects of intrastriatal injections of selective dopamine D-1 and D-2 agonists and antagonists on jaw movements of rats. AB - The effects of bilateral intrastriatal injections of the selective D-1 and D-2 antagonists, SCH23390 and sulpiride on apomorphine-induced jaw movements were studied in ketamine-anaesthetized rats after C1 spinal transection. A photo transducer attached to the lower mandible automatically detected jaw movements. Apomorphine (0.2, 0.5 and 1.0 mg/kg i.v.) dose dependently increased jaw movements, an effect prevented by prior administration into the ventral striatum of either SCH23390 (0.1, 0.5 and 1 microgram) or sulpiride (125 ng). To be effective, SCH23390 had to be given less than 30 min before apomorphine whereas sulpiride had to be given earlier. Sulpiride injected into the dorsal striatum potentiated the effects of apomorphine, an action prevented by administering the sulpiride with SCH23390. Local application of the selective D-1 and D-2 agonists, SKF38393 (5 micrograms) and quinpirole (10 micrograms) into sites within the ventral striatum from which repeated jaw movements could be obtained by electrical stimulation, also evoked jaw movements; the effects of combining the two drugs were much greater than the effects of either drug alone. PMID- 2566503 TI - Affinity chromatography with the immobilized agonist N-0434 yields an active and highly purified preparation of the dopamine D-2 receptor from bovine striatum. AB - Partial purification of the dopamine D-2 receptor from bovine striatum, solubilized in the presence of 1% digitonin, was obtained by chromatography on wheat germ lectin agarose. The preparation was purified approximately 10-fold. The stability of the receptor preparation was considerably improved and non specific protein absorption on the affinity gel used later was decreased. Further purification was achieved on a column containing a D-2-selective agonist, N-0434. Approximately 90% of the receptor activity was bound to the gel and 20-40% of the activity could be eluted by pH shock. The total purification factor after one affinity chromatography step was estimated to be at least 1500. An active preparation of at least 20% purity was obtained after a second cycle of affinity chromatography. This corresponds to an enrichment of more than 5000 times compared to the solubilized receptor preparation. PMID- 2566504 TI - Role of ancillary properties of beta-adrenoceptor antagonists in protecting the heart from anoxia. AB - The mechanism of the cardioprotective action of beta-blocking drugs against anoxia or ischemia is still not clear. We used beta-blockers (5 x 10(-7) M) of various pharmacodynamic profiles in a model of isolated, perfused working guinea pig heart subjected to 20 min of anoxia to study this. The cardioprotective effects were evaluated by measuring the recovery of the flow indices after 15 min of reoxygenation. There was a significant cardioprotective action (as measured by the effect on stroke volume recovery and on recovery of other flow and work indices) of the beta-blocking properties (nadolol, P less than 0.05), of the membrane-stabilizing property [+)-propranolol, P less than 0.05) and of a combination of these two properties with (+/-)-propranolol, which had a significantly greater effect than nadolol (P less than 0.05). The addition of weak (acebutolol) or strong (pindolol) intrinsic partial agonist activity had no clear unfavourable effect, as the degree of cardioprotection was comparable with that obtained with (+/-)-propranolol. The stroke volume recovery (percent recovery after anoxia) in the control hearts was 42.57 +/- 12.75 compared to 54.84 +/- 6.94 in hearts pretreated with nadolol, 62.99 +/- 11.41 with (+) propranolol, 71.02 +/- 11.36 with (+/-)-propranolol, 72.63 +/- 13.08 with acebutolol and 68.01 +/- 15.42 with pindolol. In vitro heart protection from anoxia with beta-blockers would appear to be related to beta-blocking activity and/or membrane stabilizing property but not a function of partial agonist activity. These ancillary properties of beta-blockers should thus be taken into account in studies on cardioprotection. PMID- 2566505 TI - Acute effects of S-adenosyl-L-methionine on catecholaminergic central function. AB - Catecholaminergic function was studied in rat brain areas after acute administration of S-adenosyl-L-methionine (SAM: 10 mg/kg i.p.). The noradrenaline (NA) concentration increased in the hippocampus and frontal cortex and decreased in the olfactory bulbs between 1 and 2 h after the SAM injection. NA biosynthesis was stimulated in vivo and the concentration of normetanephrine was increased only in the hippocampus. SAM may indirectly affect NA biosynthesis and metabolism. This effect on noradrenergic function might participate in its putative antidepressive action. PMID- 2566506 TI - Beta 2-adrenoceptor-mediated increase in the slow inward calcium current in atrial cells. AB - The effects of procaterol (beta 2-adrenoceptor agonist) on the slow inward calcium current (Isi) were examined in single cells of guinea-pig heart. Procaterol increased Isi in atrial cells, in the presence of atenolol (beta 1 adrenoceptor antagonist). The effect was abolished by ICI-118551 (beta 2 adrenoceptor antagonist). However, procaterol did not modify the membrane currents in ventricular cells. These results suggest that beta 2-adrenoceptor agonists bind to beta 2-adrenoceptors in atrial cells and augment Isi, but beta 2 adrenoceptors are not present in ventricular cells. PMID- 2566507 TI - Rilmenidine selectivity for imidazoline receptors in human brain. AB - The selectivity of three centrally acting antihypertensive agents for the medullary imidazoline-preferring receptors (IPR) versus cortical alpha adrenoceptors was investigated in human brain. [3H]Clonidine binding was studied in various membrane preparations. Competition experiments were performed. Cortical membrane preparations were used as they mainly contained classical alpha adrenoceptors whereas medullary membrane preparations from the nucleus reticularis lateralis contained only IPR insensitive to catecholamines. Rilmenidine, a new antihypertensive agent, appeared 2.5 and 3.5 times more selective than clonidine and guanfacine, respectively, for medullary IPR sites than for cortical alpha-adrenoceptors, thus providing a possible explanation for the low sedative effects of this new molecule. PMID- 2566508 TI - Brain dialysis provides evidence for D2-dopamine receptors modulating noradrenaline release in the rat frontal cortex. PMID- 2566509 TI - Quantitation and subcellular localization of proliferating cell nuclear antigen (PCNA/cyclin) in oocytes and eggs of Xenopus laevis. AB - Proliferating cell nuclear antigen (PCNA/cyclin) is a 36-kDa polypeptide present in the nuclei of mitotically active cells. It is known to be involved in DNA replication through an association with DNA polymerase delta. We examined the total content as well as the subcellular distribution of PCNA in the oocyte and the egg of Xenopus laevis by employing immunocytological staining and immunoblot analysis. While oocytes are not capable of replicating chromosomes, PCNA is abundant in the nucleus (about 65 ng per nucleus). The oocyte cytoplasm, on the other hand, does not contain a significant quantity of this protein. The amount of total PCNA does not change appreciably during oocyte maturation and the subsequent stages of egg cleavage. Thus, PCNA belongs to a class of proteins which are stockpiled during oogenesis in order to be utilized later for early embryogenesis. PMID- 2566511 TI - Temperature compensation in an ultradian rhythm of tyrosine aminotransferase activity in Euglena gracilis Klebs. AB - Tyrosine aminotransferase activity of Euglena oscillates with an ultradian period of approximately 4-5 h. The oscillation frequency in the time series was determined by cosine fitting. Experiments which were performed between 16 and 31.5 degrees C revealed temperature compensation. PMID- 2566510 TI - Ethanol ingestive behavior as a function of central neurotransmission. AB - Uncontrollable alcohol ingestive behavior has been linked to deficits of central neurotransmission. The pineal gland plays an important role in modulating ethanol intake in numerous animal species. The opioidergic (i.e. beta-endorphin, enkephalin, and dynorphin) system is involved in both the actions of alcohol and opiates, as well as craving and/or genetic predisposition towards abuse of these two agents. Furthermore, there is significant evidence to link ingestive behaviors with the ventral tegmental accumbens-hypothalamic axis, whereby the biogenic amines dopamine and serotonin are reciprocally involved. Evidence is presented which implicates the striatum and the hypothalamus as possible specific loci for regional differences between alcohol-preferring and alcohol nonpreferring mice. We believe that photoperiod-induced alcohol ingestive behavior may involve alterations in both pineal and hypothalamic opioid peptides. PMID- 2566512 TI - New expression vectors for the fission yeast Schizosaccharomyces pombe. AB - A general expression vector (pMB332) for the fission yeast Schizosaccharomyces pombe was constructed. The heterologous gene expression is driven by the S. pombe alcohol dehydrogenase (adh) promoter. Transcription termination signals were isolated from the S. pombe actin gene. The vectors carry the Saccharomyces cerevisiae Ura3 gene, which complements the S. pombe ura4 mutation. The plasmid stability is conferred by the S. pombe ars and stb elements isolated from pFL120 [(1983) Cell 32, 371-377]. An 'ATG' vector (pMB340) was created, which allows the expression of protein fragments fused to a translational start codon downstream of the adh promoter. The function of this vector system is shown by the production of the human blood coagulation protein factor XIIIa. PMID- 2566513 TI - [The role of the workers of a feldsher-midwife center in the battle against infertility]. PMID- 2566515 TI - [Blood-sucking flies and the protective measures against them]. PMID- 2566514 TI - [The role of the feldsher in the prevention and treatment of trauma among agricultural workers]. PMID- 2566516 TI - [The organization of therapeutic and prophylactic care for children at feldsher midwife centers]. PMID- 2566517 TI - [Gamma-aminobutyric and glutamic acid levels in the brain of rats exposed to noise and vibration on ships]. AB - The influence of different levels of noise and vibration on the content of GABA and glutamic acid in the brain and on behavioural characteristics of rats have been studied on 60 white rats under voyage conditions. A correlation is determined between biochemical and physiological reactions of the central nervous system in animals and duration and level of the influence of the studied factors. PMID- 2566518 TI - [The role of the endothelium in the reactions of the vascular wall to mechanical stretching and the action of neuromediators]. AB - In isolated rings of the rat aorta, the responses of the smooth muscles to mechanical stretching were studied: the endothelium cells potentiated the relaxation under the stretching. After removal of endothelium cells, acetylcholine induced contraction of the smooth muscle. PMID- 2566519 TI - Lethal course of X-linked dominant chondrodysplasia punctata in a male newborn. AB - We report a newborn with some manifestations of chondrodysplasia punctata. Additional abnormalities were hydrocephalus, bilateral syndactyly of the fourth and fifth fingers and toes, absence of the middle phalanx of all toes, hypoplasia of the second and third phalanges of all fingers and cryptorchidism. This observation suggests that we are possibly dealing with a rare male case of X linked dominant chondrodysplasia punctata. PMID- 2566521 TI - Prospects for nonsurgical female sterilization. PMID- 2566520 TI - Effects of small variations in insulin and glucagon levels on plasma aminoacids concentrations. AB - To determine the effect in normal subjects of small variations of insulin and glucagon on plasma aminoacids concentrations we suppressed endocrine pancreas secretion with somatostatin and measured aminoacids levels during a sequential insulin infusion in the absence (control test, low glucagon level) or in the presence (normal glucagon concentration) of a replacement glucagon infusion. Insulin infusion rates were 0.05, 0.09, 0.15 and 0.30 mU.kg-1.min-1 during the control test and 0.09, 0.15, 0.30 and 0.40 mU.kg-1.min-1 during the replacement test. During the control test, glucagon decreased (p less than 0.01) and insulin levels were successively 8.2 +/- 0.4, 10.1 +/- 0.7, 11.9 +/- 0.14 and 18.5 +/- 0.8 mU.l-1. The only effect on insulin was to decrease branched-chain aminoacids (BCAA). BCAA were inversely related to insulinemia (p less than 0.01). A significant decrease was obtained for an insulin level of 11.9 +/- 0.4 mU.l-1, a value intermediate between those decreasing glycerol (10.1 +/- 0.7 mU.l-1) and stimulating total body glucose uptake (18.5 +/- 0.8 mU.l-1). During the test with glucagon replacement glucagon was maintained at its initial value. Insulin levels were successively 8.3 +/- 0.3, 11.9 +/- 0.3, 19.7 +/- 0.6 and 26.7 +/- 0.5 mU.l 1. Insulin decreased always BCAA but also threonine, proline, tyrosine, methionine and total aminoacid levels. BCAA were always inversely related to insulin levels (p less than 0.01) but the slope of the relationship was modified and more insulin was needed to decrease BCAA concentration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566522 TI - Evaluation of the benefit of setting up a decentralised obstetric-surgical structure in order to reduce maternal mortality and transfers in the Ivory Coast. AB - Our purpose is to study the way to reduce maternal mortality when introducing a structured obstetric and surgical center to a community previously without such a center. The study was done in Ivory Coast, where such a structure was set up in 1982 in a rural hospital at Divo. It has been observed that with such a structure there are no more transfers of women from Divo to the teaching hospital at Cocody, and a reduction in maternal mortality at Divo. This observation regarding the value of bringing obstetric services to communities currently without them could be a way among those available to reduce maternal mortality in a developing country such as the Ivory Coast. PMID- 2566523 TI - Zinc and copper concentration in plasma of pregnant women in Nigeria. AB - The concentration of plasma zinc and copper were determined by atomic absorption spectrophotometry in 58 Nigerian women at various times in pregnancy and compared with those of 11 non-pregnant controls. The results showed 67.5% decrease in zinc and 100% increase in copper over non-pregnant levels at the end of pregnancy. The mean plasma zinc levels were appreciably lower and copper the same as published values for Caucasian women at similar periods in pregnancy. Furthermore, there was no correlation between plasma zinc and copper and neonatal birthweight. We conclude that (1) a significant proportion of pregnant women in Nigeria have biochemical hypozincemia; (2) this hypozincemia is not responsible for fetal growth retardation in this population and (3) there is at present no justification for giving routine zinc supplementation to pregnant women in Nigeria. PMID- 2566524 TI - Umbilical artery velocity waveforms are not valid indices for assessing growth retardation in utero. AB - Blood flow velocity waveforms from the umbilical artery of 126 normal fetuses with no complications at 20-41 weeks and in 31 growth-retarded fetuses, were recorded using pulsed Doppler ultrasound. A total of 140 examinations were performed on normal fetuses and 49 on growth-retarded fetuses. The resistance index (RI) was used as an index of blood flow resistance. In 30 of the 31 growth retarded fetuses (96.8%), the values of RI were within the mean +/- 2 S.D. of normal fetuses. Placental/fetal weight ratio (P/F ratio) was assessed. All values of P/F ratio of growth-retarded fetuses were within the mean +/- 2 S.D. of normal values. Small placentas expressed as a normal P/F ratio almost always associated with normal RI values. Therefore, measurement of RI on umbilical artery blood flow velocity waveform by Doppler ultrasound cannot serve as a valid diagnostic index to predict growth retardation of the fetus in utero. PMID- 2566525 TI - Umbilical cord gamma-glutamyl transpeptidase and placental dysfunction. AB - Placental dysfunction has been suspected if human placental lactogen level and/or cystine-aminopeptidase activity were lower than 10th centile. Significant rise in gamma-glutamyl transpeptidase (GGTP) has been found in samples of arterial cord blood of newborns born to mothers with placental insufficiency. We observed the relationship between GGTP activity and decrease in cord pH and lower Apgar score of corresponding infants. We suggest that chronic hypoxemia as a consequence of placental dysfunction may result in damage of fetal liver cells. PMID- 2566526 TI - Fetal foot length--a new parameter for assessment of gestational age. AB - Ultrasonographic measurement of fetal foot length, a new parameter, was correlated with the gestation age. One hundred and five ultrasonographic measurement of fetal foot length was performed between 13 and 42 weeks gestation. Comparison of linear regression of foot length versus gestational age demonstrated a strong correlation with an r2 value of 0.84 (P less than 0.001). Ninety-five percent confidence intervals at each week compared favorably with both biparietal diameter and femur length data. Mean foot lengths at each week of gestation compared favorably with data based on pathological specimens described in 1920 (Streeter GL: Weight, sitting height, head size, foot length and menstrual age of the human embryo. Contrib Embryol Carnegie Inst. 11: 143, 1920). Measurement of fetal foot length is of particular use when other parameters do not accurately predict gestational age, e.g. hydrocephalus, anencephaly, short limb dysplasia. It can also be used in conjunction with biparietal diameter and femur length in the management of patients with premature labor in order to patients with premature labor in order to accurately predict gestational age. Hence the present study demonstrates that the ultrasonographic measurement of foot length is a reliable indicator of gestational age. PMID- 2566527 TI - Screening for toxoplasmosis in pregnancy. AB - Randomly collected sera from 386 pregnant women attending obstetric and gynecology clinics at Kind Khalid University Hospital, Riyadh, Saudi Arabia, were examined for toxoplasma antibodies by five serological methods, i.e. latex agglutination test (LAT), two indirect hemagglutination tests (IHAT) (Carter Wallace, USA and Ismunit, Italy), enzyme immunoassay (EIA) and indirect fluorescent antibody test (IFAT). The percentage of sensitivity, specificity and coincidence value of these tests were compared with IFAT which was used as a reference test. For routine screening of toxoplasmosis, LAT has proved in this study to be the most suitable test. The LAT is cost effective and easy to perform. In this study of the three tests (IFAT, EIA, immunosorbent agglutination assay) to demonstrate specific IgM for toxoplasmosis, the EIA test proved to be the most satisfactory because of its 99% specificity. If EIA equipment is available, it can be used for routine screening (IgG) as well as IgM determination. The incidence of toxoplasmosis in pregnant women varied between 25.4% and 36.3% depending on the method used. PMID- 2566528 TI - Biomagnetic measurements in uterine leiomyomas using a superconducting quantum interference device (SQUID). AB - Using the biomagnetometer superconducting quantum interference device (SQUID), the magnetic signals relating to spontaneous uterine activity were measured for the first time in 25 women with uterine leiomyomas, and 12 women with normal, non leiomyomatous, uteri. Magnetic radiation, in the range of low frequency (below 2 Hz), was of low amplitudes in normal uterine tissues and of high amplitudes in benign leiomyomas. The observed differences were confirmed by the corresponding power spectra obtained from the statistical Fourier analysis. It is suggested that biomagnetic measurements may facilitate the detection of uterine leiomyomas in equivocal cases. PMID- 2566529 TI - The use of prostaglandins and antiprogestins for pregnancy termination. AB - Although vacuum aspiration could be regarded as a simple procedure, complications do occur and attempts have been made to develop non-surgical procedures for termination of pregnancy during the first 3-4 weeks following the first missed menstrual period. A variety of prostaglandin (PG) analogs have been developed which are equally effective as vacuum aspiration to induce abortion during early pregnancy. However, the widespread acceptance of PG treatment is limited by a relatively high incidence of gastrointestinal side effects and uterine pain. Treatment with presently available antiprogestins alone is not sufficiently effective to compete with vacuum aspiration. However, administration of these compounds induces uterine contractions and increases the sensitivity of the myometrium to prostaglandin. These effects allow the development of sequential treatment with RU 486 and a low dose of PG analogs administered intramuscularly or vaginally which is highly effective and is seemingly associated with a lower frequency of side effects than if PG analogs are used alone. It can be concluded that this medical abortion method has the capacity to compete with vacuum aspiration for termination of early pregnancy. Randomized studies comparing the two procedures are, however, needed to confirm this statement. PMID- 2566530 TI - Genital herpes simplex virus infections: laboratory confirmation in diverse patient groups. AB - The laboratory diagnosis of genital herpes simplex virus (HSV) infections in various patient groups with diverse clinical manifestations including asymptomatic pregnant women with history of genital herpes and in a high prevalence group consisting of male patients with recurring genital ulcerations is described. HSV was detected by conventional cell culture and also by a CPE enhancement technique using human embryonic lung fibroblasts (MRC-5) and Vero cells with subsequent typing of isolate by HSV type-specific monoclonal antibodies. A definite improvement in HSV diagnosis was noted by the use of the CPE-enhancement technique with the reduction of turn-around time to 24 h at which 61.1% and 50% isolation rates were detected in MRC-5 cells in comparison to 38.9% and 16.6% by conventional cell culture in the high prevalence patient population and asymptomatic pregnant women, respectively. Delayed development of CPE and reduced isolation rates were detected using Vero cells. Follow-up sampling of subsequent episodes of genital ulcerations in those with history of recurrence but an initial negative cell culture result, increased the isolation rate substantially and enabled the confirmation of diagnosis of genital HSV infection. PMID- 2566532 TI - A 15-year study of illegally induced abortion mortality at Ilorin, Nigeria. AB - Fifty-three women died from complications of illegally induced abortion over a 15 year period. Seventeen (32.2%) of them were under 20 years of age, while 28 (52.8%) were nulliparous. The abortion rate was 210/100,000 with an abortion mortality rate of 0.9/100,00 women. Also the abortion ratio was 73.4/1000 live births while the abortion mortality ratio was 4.20/1000 abortions. Sepsis and hemorrhage were the two commonest avoidable causes of death, and preventable measures were discussed. PMID- 2566531 TI - Ovarian development of the female child and adolescent: I. Morphology. AB - Ovarian morphology of 180 randomly selected females, aged 8-21, was investigated, utilizing standard sector scanning ultrasound techniques, at -1 year (1 year before menarche), year 0 (menarche) and years +1 to +8 (1-8 years postmenarche). According to strict morphologic criteria, seven ovarian patterns were recognized which varied in dominance with age in reference to menarche. Prior to menarche a multifollicular ovarian pattern (Type 1) dominates which after menarche is substituted by a predominantly active ovarian pattern (Type 5). Approximately 20% of postmenarcheal females do not develop this active pattern. In a relatively persistent percentage of females a microcystic (Type 2) ovarian pattern was recognized throughout all postmenarcheal years, suggestive of a normal transitional pattern from Type 1 to Type 5 ovaries. In contrast, multicystic (Type 3), hyperthecosis (Type 4), polycystic ("Mickey Mouse") (Type 6) and silent (Type 7) ovaries appear to represent abnormal ovarian developments. Based on percentage distributions, it is tempting to speculate that multicystic (Type 3) and/or hyperthecosis (Type 4) ovaries represent precursor ovaries to Type 6 (polycystic) ovaries. Confirmation of this hypothesis would have major clinical importance for the early diagnosis of the polycystic ovarian syndrome. PMID- 2566534 TI - Retained pregnancy complicating donor ovum transfer. AB - Retained pregnancy is an uncommon complication of females participating in uterine lavage for purposes of ovum donation. Reported are two subjects experiencing this complication. Unlike other ovum donors, uterine retroflexion was present in both women. This anatomic configuration was likely responsible for the retained pregnancies and such women are best excluded from participating in this method of embryo donation. PMID- 2566533 TI - The effect of cervical dilatation by laminaria tent in first trimester legal abortions on blood loss related to fibrinolytic activity in the decidua and placenta. AB - Increased blood loss (BL) has been reported after cervical dilatation by laminaria tent in legal abortions. The BL was measured in 72 women in whom first trimester legal vacuum aspiration was performed. The cervical canal was dilatated by laminaria tent in 37 patients, and mechanically with Hegar dilators in 35 patients. BL was studied in relation to the plasminogen activators (u-PA, t-PA) and the plasminogen activator inhibitors (PAI-1, PAI-2) in the decidua and placenta. There was no significant difference in BL between the two groups. Decidual PAI-1 concentrations were significantly higher in the laminaria tent group than in the Hegar dilator group. An inactivation of u-PA by PAI-1 might explain the lack of increase in BL among the laminaria tent group. PMID- 2566535 TI - Virilizing granulosa cell tumor responsive to human chorionic gonadotropin and oral contraceptive with 8-year followup. AB - Urinary and serum steroid measurements were obtained in a 48-year-old female before and after the removal of a virilizing granulosa cell tumor (GCT). Preoperatively, serum testosterone was markedly elevated while 17-ketosteroid (17 KS) and estrogen excretion were normal. Chorionic gonadotropin administration effected a marked rise in urinary 17-KS excretion and serum testosterone while oral contraceptive therapy for 21 days decreased serum testosterone to normal. With tumor resection, serum testosterone fell to normal and subsequent administration of HCG had no stimulatory effect on serum testosterone or urinary 17-KS excretion. The patient's clinical response and 8-year followup attest to the tumor's unusual virilizing role. PMID- 2566536 TI - Comparative efficacy of once daily loratadine versus terfenadine in the treatment of allergic rhinitis. AB - This 1 week study compared the efficacy of once daily administration of 10 mg loratadine with 120 mg terfenadine in out-patients with seasonal allergic rhinitis. It focussed on the efficacy of treatment at the end of the 24 h period following a daily dose. The study was designed as a double-blind, randomized, parallel-group trial, and 41 patients were enrolled and evaluated for efficacy. Patients took an initial dose at the study site and returned on days 2 and 8. At day 2 (24 h after the initial dose), according to the physician's evaluation 57% of loratadine-treated patients had a good or excellent response, compared to 50% of those given terfenadine. At day 8, 24 h after the final dose, 71% of the loratadine-treated patients and 35% of the terfenadine-treated patients had a good or excellent response (P = 0.03). At days 2 and 8, reductions in mean symptom scores measured 22, 23 and 24 h after the initial and final doses showed an indication of being greater with loratadine than with terfenadine (nonsignificant due to small sample size). The incidence of sedation was similar in both groups. It is concluded that 10 mg loratadine, administered once daily, controls the symptoms of rhinitis more effectively than 120 mg terfenadine given once daily in the last few hours of the 24 h dosing period. PMID- 2566537 TI - A study of the anticholinergic activity of terfenadine in normal volunteers. AB - The anticholinergic effects of single doses of terfenadine, chlorpheniramine and atropine on salivary flow were compared with those of placebo in a double-blind crossover study in 21 normal adults. Salivary flow was measured immediately before administering study medications and 3 h after dosing, for the evaluation of anticholinergic activity. Mean salivary flow decreased significantly after administration of chlorpheniramine and atropine (P less than 0.01 and P less than 0.001, respectively), and was unchanged by terfenadine and placebo, indicating that terfenadine does not possess detectable anticholinergic activity. Mild drowsiness was reported by 57% and 29% of subjects who received chlorpheniramine and terfenadine, respectively. PMID- 2566538 TI - A review of developments in H2-receptor antagonist therapy: focus on famotidine. Proceedings of an official satellite symposium to the 8th Asian-Pacific Congress on Gastroenterology and the 5th Asian-Pacific Congress of Digestive Endoscopy. Seoul, Korea, 10 October 1988. PMID- 2566539 TI - Famotidine in the USA: a review of efficacy studies. AB - Since its introduction into the USA, famotidine has been widely studied. A variety of studies has shown it to be a very potent H2-receptor antagonist; more potent and longer acting than cimetidine or ranitidine. It has been shown to be efficacious, cost-effective and relatively safe in the treatment of duodenal ulcers, benign gastric ulcers, in maintenance therapy, and when used intravenously in the critical care setting. PMID- 2566540 TI - Recent advances in the treatment of digestive diseases. AB - Significant advances in the treatment of peptic ulcers have occurred during the last 25 years. Two important problem areas remain: resistant ulcers and ulcer relapse. Ulcers that remain unhealed after 8 weeks of treatment with full doses of active anti-ulcer drugs are considered resistant. Risk factors for the development of resistance are known, the most important being non-compliance with recognized effective therapy. A correlation exists between healing of duodenal ulcers and degree of gastric acid suppression. A duodenal ulcer that fails to heal with mild suppression of gastric acidity may, therefore, respond to more potent and longer acting drugs, such as famotidine or omeprazole. Prescribing drugs that may improve the mucosal defence may also be useful. For gastric ulcers, prolonging treatment may be effective. Currently available drugs provide some protection from ulcer relapse, but the risk of an ulcer recurring remains. Most recurrences are asymptomatic and ulcers are most likely to recur within a relatively short period of time. Success in the treatment of resistant ulcers and maintenance therapy to prevent relapse may require greater understanding of the pathology of ulcers and application of drug therapy that specifically corrects these abnormalities. PMID- 2566541 TI - Famotidine in the treatment of duodenal ulcers resistant to other histamine H2 receptor antagonists. AB - In 20% of patients with duodenal ulcers, treatment is ineffective. Ulcers are considered resistant when they fail to heal following 8-12 weeks of treatment with full dose duodenal ulcer therapy. The present study reviewed the records and endoscopic findings of 73 patients who were considered by their physicians to have resistant duodenal ulcers. From this population, 15 patients with documented, active duodenal ulcers were chosen to participate in this study. Each had failed treatment with cimetidine or ranitidine at recommended doses administered for from 56 days to more than 2 years. After up to 6 weeks' treatment with 20 mg famotidine given twice daily, eight patients showed endoscopic evidence of healing (S1 or S2) based on the criteria established by the Japanese Society of Digestive Endoscopy. Five others showed moderate to marked improvement H1-H3). No patient experienced side-effects during famotidine treatment. The greater antisecretory effect of famotidine may be useful in treating duodenal ulcer patients who fail to heal with cimetidine or ranitidine. PMID- 2566542 TI - Treatment of upper gastro-intestinal bleeding with the H2-receptor antagonist famotidine. AB - Gastro-intestinal bleeding from peptic and stress ulcers is serious and life threatening. Critically ill patients in intensive care units have many of the risk factors associated with bleeding from peptic and stress ulcers, including trauma, burns, sepsis, shock and multiple organ failure. This study investigated the results of treatment with famotidine, administered intravenously twice daily, to those in a control group that received treatment before the introduction of H2 receptor antagonists. The study was designed to determine whether famotidine reduced the need for emergency surgery in patients with bleeding ulcers and whether a reduction in mortality was associated with its use. The overall efficacy rate of famotidine was greater than 88%. The percentage of patients with a bleeding ulcer undergoing surgery was 24.5% compared with 50.3% in the historical control group. Twice daily intravenous administration of famotidine effectively stopped bleeding in patients with moderate to severe peptic ulcer and stress ulcer. Drug therapy for the treatment of upper gastro-intestinal bleeding, however, has limitations. Criteria for the use of famotidine include reduced mortality, rate of recurrent bleeding and rate of emergency surgery. PMID- 2566543 TI - Safety profile of H2-receptor antagonists: current status of famotidine. AB - An assessment of the adverse reactions associated with H2-receptor blockers is presented, with particular emphasis towards famotidine. Post-marketing studies on the H2-receptor blockers have shown that they have a low incidence of adverse reactions (1-2% of patients) and are very effective in the treatment of peptic ulcers. The most common adverse reactions are gastro-intestinal, although central nervous system, cardiovascular, haematological, endocrine and hepatic disturbances may occur. Drug interactions and the considerable amount of post marketing surveillance data, spanning over a decade of use, are also assessed. PMID- 2566544 TI - Pharmacology of H2-receptor antagonists: an overview. AB - The H2-receptor antagonists which are used for ulcer therapy fall into four main structural classes: cimetidine is an imidazole derivative; ranitidine and nizatidine belong to the basically substituted furans and thiazoles, respectively; famotidine is a member of the guanidino-thiazole group; and roxatidine belongs to the aminoalkylphenoxy series. Famotidine is the most potent, selective H2-receptor antagonist yet available for ulcer therapy. On a weight basis, famotidine is approximately eight times more potent than ranitidine and 40 times more potent than cimetidine. Furthermore, it increases the gastric mucosal blood flow, resulting in an increased haemostatic effect. Famotidine has a longer duration of action than either ranitidine or cimetidine. Since famotidine does not interact with cytochrome P-450 of the hepatic enzyme system, it does not appear to affect the metabolism of drugs metabolized by this system. The overall incidence of side-effects of the H2-receptor antagonists is in the range of 2-3% and no irreversible adverse effects are known. Famotidine has been found to be generally well tolerated. In post-marketing studies, the percentage of patients with side-effects is in the range 1.2-2%. Thus, famotidine is a safe and potent H2-receptor blocker of acid secretion. PMID- 2566545 TI - Molecular and clinical characterization of an insertional polymorphism of the insulin-receptor gene. AB - A restriction-fragment-length polymorphism (RFLP) detected with the human insulin receptor cDNA and the enzyme Sac I has been reported to be associated with non insulin-dependent diabetes mellitus (NIDDM) in White and Black populations and segregated with diabetes in two small pedigrees with maturity-onset diabetes of the young. A size difference of approximately 500 base pairs (bp) was demonstrated between the alleles of this and several other RFLPs that mapped to the same 300-bp region near the transmembrane coding region of the cDNA beta chain, thus suggesting the presence of an insertion in this region that could affect insulin-receptor function. Genomic DNA fragments containing this RFLP were cloned from an individual heterozygous for the putative insertion, and the differing fragments of the two alleles were sequenced. The presence of a 400-bp insertion was thus confirmed and was demonstrated to be entirely within an intron. No significant coding-region differences from published cDNA sequences were detected in four exons sequenced from the region of the insertional allele. The sequenced regions included multiple Alu repeat sequences. The RFLP was unusual in that the larger allele consisted of an additional Alu repeat sequence that included a new Pst I site. Because the nature and location of the insertion did not suggest a role in insulin-receptor function, the association of this RFLP with NIDDM and hyperinsulinemia was reexamined in a small sample of Whites. No association could be demonstrated, and the insertion also failed to segregate with NIDDM in five White pedigrees.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566546 TI - Exercise-induced rise in glucagon and ketogenesis during prolonged muscular work. AB - These experiments examined the role of the exercise-induced increment in glucagon in the control of ketogenesis during prolonged moderate-intensity (100 m/min, 12% grade) treadmill exercise. Dogs were studied during 150 min of exercise with saline infusion alone (C; n = 6) with the glucagon levels clamped at basal values (somatostatin infusion with basal glucagon replacement and the normal fall in insulin simulated; BG; n = 5) or with the normal exercise-induced rise in glucagon simulated (somatostatin infusion with the rise in glucagon and the fall in insulin simulated; SG; n = 5). Glucose was infused as needed in SG and BG to maintain the glycemic response seen in C. In all dogs, catheters were inserted into the carotid artery and the portal and hepatic veins for blood sampling and the vena cava and the splenic vein for infusions. Glucagon rose from 62 +/- 5 and 57 +/- 4 pg/ml at rest to 104 +/- 20 and 120 +/- 12 pg/ml during exercise in C and SG but did not deviate from basal in BG (56 +/- 3 pg/ml). Insulin fell similarly from rest to the end of exercise in C (13 +/- 2 to 5 +/- 1 microU/ml), SG (11 +/- 1 to 6 +/- 1 microU/ml), and BG (10 +/- 1 to 6 +/- 1 microU/ml). In C, SG, and BG, free-fatty acid (FFA) levels rose from 941 +/- 81, 1240 +/- 155, and 938 +/- 36 mu eq/L at rest to 1615 +/- 149, 1558 +/- 175, and 1391 +/- 160 mu eq/L with exercise.2+n C, PMID- 2566547 TI - Restriction fragment length polymorphism analysis of major histocompatibility complex genes in the non-obese diabetic mouse strain and its non-diabetic sister strains. AB - It has been suggested that one of the recessive genes controlling diabetes in non obese diabetic mice is linked to the major histocompatibility complex. We, therefore, performed restriction fragment length polymorphism studies of major histocompatibility complex genes (class I, II, and III) in non-obese diabetic mice in comparison with those of their non-diabetic sister strains, non-obese non diabetic, cataract, and ILI mice which were derived from the same Jcl-ICR mice as the non-obese diabetic mouse was. When class II and III probes and a minimum of four restriction enzymes were used, class II and III genes of non-obese diabetic mice were indistinguishable from those of cataract and ILI mice but totally different from those of non-obese non-diabetic mice. The studies also indicated that A beta, E beta, and C4-Slp genes of non-obese diabetic, cataract, and ILI mice, and A alpha, A beta, E beta and C4-Slp genes of non-obese non-diabetic mice are different from those of BALB/c and C57BL/6 mice, respectively. While non obese non-diabetic mice expressed the E alpha gene, non-obese diabetic, cataract, and ILI mice appeared to carry a deletion in the 5' end of the E alpha gene resulting in failure to transcribe the E alpha gene. When class I probe was used, cataract mice showed very different band patterns from those of the other ICR derived mice. It is suggested that non-obese diabetic, non-obese non-diabetic, and ILI mice contain only a single class I D region gene.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566548 TI - Effect of alpha a-adrenoceptor antagonist on platelet activation during insulin induced hypoglycaemia in type 2 (non-insulin-dependent) diabetes mellitus. PMID- 2566549 TI - [Prevention of the first hemorrhagic complication by rupture of esophageal varices in cirrhosis]. PMID- 2566550 TI - Neurohumoral pathways mediating stress-induced changes in rat gastrointestinal transit. AB - Pharmacologic and surgical procedures were used to define the neurohumoral pathways that mediate stress-induced changes in gastrointestinal transit in the rat. Physical restraint (a validated stressor) resulted in significant inhibition of gastric emptying and small bowel transit but in stimulation of large bowel transit. Most importantly, the results indicate that stress-induced changes in gastrointestinal transit in rats are differentially mediated by sympathetic and parasympathetic efferents of the autononic nervous system and, in part, by opiate pathways but not by the pituitary and adrenal glands. Stress-induced inhibition of gastric emptying is mediated by noradrenergic efferents, whereas stress induced changes in small and large bowel transit are predominantly mediated by vagal, parasympathetic efferents. PMID- 2566551 TI - Colonic N-acetylation of 5-aminosalicylic acid in inflammatory bowel disease. AB - 5-Aminosalicylic acid presently is believed to represent the therapeutically active moiety of the sulfasalazine molecule in the treatment of inflammatory bowel disease. The metabolism of this compound, however, has not been studied in detail. In this paper we provide evidence that 5-aminosalicylic acid is acetylated to N-acetyl-aminosalicylic acid in homogenates from colonic biopsy specimens (370 +/- 20 nmol/g wet wt or 2.9 +/- 0.9 nmol/mg.min, n = 10), whereas acetylation in fecal samples was only small (13.0 +/- 3.0 nmol/g). Mucosal N acetylation was rapid, cofactor- and pH-dependent, and could be enriched in the cytosolic fraction. In contrast, fecal acetylation was slow and did not depend on the presence of acetyl-coenzyme A. There were neither significant differences of acetylation between patients and controls nor a significant correlation to the individual acetylation phenotype. From our results we believe that presystemic acetylation of 5-aminosalicylic acid may be mainly mediated by a colonic mucosal enzyme and only to a small extent by fecal (bacterial) processes. PMID- 2566553 TI - [Characteristics of lymphocyte autoantigens]. AB - Specificity of autoantibodies against lymphocyte antigens was assayed in sera of patients with autoimmune hematologic diseases, leukemias, secondary immunodeficiencies as well as in sera of persons seropositive to antibodies against retroviruses HTLV-I and HTLV-III. Autoantigens of the lysates of stimulated peripheral blood lymphocytes and Hut-102 cells were characterized with the aid of immune blotting and radioimmunoprecipitation. No differences were found in autoantigen spectra identified by sera from patients belonging to different groups. Autoantigens 75 cd, 60 cd, 50 cd and 12 cd occurring in both cell types were demonstrable at a greater frequency. Some of the sera also demonstrated the total antigens 34 cd and 29 cd. The role of autoantigens in the pathology development is discussed. PMID- 2566552 TI - [The immunohistochemical growth fraction (Ki-67) of breast cancers: relations to tumor spread, tumor morphology and receptor testing]. AB - 245 breast cancer tissues were analysed immunohistochemically, using monoclonal antibody Ki-67, which spezifically reacts with a nuclear antigen of proliferating cells and represents the so-called growth fraction. According to the number of positive cells, three Ki-score categories (I-III) were established. The results were compared with prognostic variables (Histological grading, DNA cytophotometry), staging parameters, immunocytochemical (ER-ICA, mPRI), and biochemical (DCC) hormone receptor assay. We found a good correlation between Ki 67 and grading (Bloom and Richardson): G 1-tumors had small growth fractions, in contrast to G-III carcinoma, which showed a high percentage of Ki-Score III. Receptor negative cases had a large growth fraction and receptorpositive ones showed an uniform distribution. The comparison between Ki-67 and tumor spread or DNA-Cytophotometry showed no correlation. The majority of 31 recurrent tumors not only had a large number of positive cells, but also a definite dependence was found between Ki-Score and length of the disease-free period. Ki-67 seems to be an important immunohistochemical marker of breast cancer, which contributes to the individual assessment of the disease and can possibly give predictions on tumor development and response to radiation and systemic therapy. PMID- 2566555 TI - Role of glutamine as a direct co-repressor of glutamine synthetase in Rhodobacter capsulatus E1F1. AB - High performance liquid chromatography (HPLC) has been used to determine the internal levels of amino acids in Rhodobacter capsulatus E1F1 cells, subjected to different treatments and nutritional conditions. Glutamine synthetase activity and enzyme concentration correlated negatively with the level of glutamine, suggesting that glutamine per se acts as a co-repressor in the enzyme synthesis. Moreover, addition of the specific inhibitor L-methionine-D,L-sulfoximine, that produced an increase in enzyme concentration, specifically promoted a depletion of intracellular glutamine. PMID- 2566554 TI - Transport of nutrients into the renal brush border membrane vesicles as marker in evaluating the role of antipili antibodies in modulation of ascending pyelonephritis in rats. AB - The uptake of D-glucose, L-aspartate, L-lysine and L-proline was investigated in renal brush border membrane (BBM) vesicles prepared from control, infected or passively-immunized-infected rats. Except L-aspartate, a progressive decrease in the uptake of these nutrients in both infected and immunized-infected groups during the course of infection was observed, but the changes were less apparent in immunized-infected rats than in non-immunized ones. The uptake of L-aspartate was increased in vesicles from early stages of infection but decreased in those from later stages. Also in L-aspartate uptake, the changes were smaller in immunized animals. The uptake of nutrients was detectable earlier than were histopathological alterations of both kidneys. The observations demonstrated that uptake of D-glucose and amino acids in the kidneys is disturbed prior to appearance of histopathological lesions and thus can be used for early detection of the disease. The data also demonstrate that antipili antibodies afford partial protection against ascending pyelonephritis. PMID- 2566557 TI - Haplotype-specific interactions of non-H-2-linked genetic factors controlling the mouse C4 and Slp protein levels. AB - The influence of non-H-2 linked genes on the plasma levels of the H-2 S-region encoded proteins C4, Slp, and factor B was tested in Recombinant Inbred (RI) strains. The A X B and B X A RI strains exhibit a continuous range of C4 and Slp levels from very high to very low which reach beyond the levels of their parental strains, C57BL/6J and A/J, indicating involvement of several trans-regulatory (non-H-2-linked) genes. Only limited variation in levels of factor B has been found. No linkage relationship could be established for the trans-regulatory genes, because more than one gene is involved. A complex interaction of H-2 haplotype, genetic background, sex, and possibly maternal effect in determining the C4 and Slp protein plasma levels has been observed. The H-2-dependent sex effect is evident, because males have higher C4 levels than females in RI strains with H-2b but not with H-2a haplotype. This sex effect is also background dependent, because it is present in the H-2b congenic strain on A background (A.BY) but not in C57BL/10 and C57BL/6 (both H-2b). Mice from RI strains with H 2b haplotype have in general higher C4 levels than mice with H-2a haplotype. PMID- 2566556 TI - The isolation and sequence of missense and nonsense mutations in the cloned bacteriophage P22 tailspike protein gene. AB - Twenty-seven new mutations in the structural gene for the Salmonella typhimurium bacteriophage P22 tailspike protein have been isolated, mapped using a powerful plasmid-based genetic system and their DNA sequence changes determined. The mutations were generated by hydroxylamine treatment of the cloned gene on a plasmid expression vector. Assaying the activity of the tailspike protein produced from this plasmid and screening for plasmid mutants were accomplished by the in situ complementation of P22 capsids imbedded in soft agar to produce infectious phage. Deletion mutations in the cloned gene have been constructed by a two step procedure involving oligonucleotide linker insertion and in vitro deletion by restriction endonuclease digestion. The deletions, whose physical endpoints were determined by DNA sequencing, define 12 genetic and physical intervals into which the new mutations were mapped by marker rescue experiments. These deletions were transferred to phage P22 by recombination and used to map mutations carried on plasmids. Following mapping, the nucleotide change for each of the mutations was determined by DNA sequencing. The majority were absolute missense mutations although both amber and ochre nonsense mutations were also identified in the protein coding portion of the gene. The suppression pattern of the nonsense mutations was determined on several nonsense suppressors. Four of the mutations cause severely depressed levels of tailspike protein expression from both the cloned gene on the plasmid expression vector and from P22 phage carrying these mutations. These mutations were identified as nucleotide changes in what is probably the P22 late operon transcription terminator which immediately follows the tailspike protein coding sequence. PMID- 2566558 TI - Physical and genetic characterization of a 75-kilobase deletion associated with al, a recessive lethal allele at the mouse agouti locus. AB - The agouti locus (A) of the mouse determines the timing and type of pigment deposition in the growing hair bulb, and several alleles at this locus are lethal when homozygous. Apparent instances of intragenic recombination and complementation between different recessive lethal alleles have suggested that the locus has a complex structure. We have begun to investigate the molecular basis of agouti gene action and recessive lethality by using a series of genetically linked DNA probes and pulsed field gel electrophoresis to detect structural alterations in radiation-induced agouti mutations. Hybridization probes from the Src and Emv-15 loci do not reveal molecular alterations in DNA corresponding to the ae, ax, and al alleles, but a probe from the parotid secretory protein gene (Psp) detects a 75-kilobase (kb) deletion in DNA containing the non-agouti lethal allele (al). The deletion is defined by a 75-kb reduction in the size of BssHII, NotI, NruI and SacII high molecular weight restriction fragments detected with the Psp probe and is located between 25 kb and 575 kb from Psp coding sequences. Because the genetic distance between A and Emv-15 is much less than A and Psp, there may be a preferred site of recombination close to Psp, or suppression of recombination between A and Emv-15. The al deletion has allowed us to determine the genotype of mice heterozygous for different recessive lethal alleles. We find that three different recessive lethal complementation groups are present at the agouti locus, two of which are contained within the al deletion. PMID- 2566560 TI - Isolation, structure, and expression of labial, a homeotic gene of the Antennapedia Complex involved in Drosophila head development. AB - The labial (lab) gene of Drosophila melanogaster is necessary for the proper development of the embryonic (larval) and adult head. We have identified the lab transcription unit within the proximal portion of the Antennapedia Complex (ANT C) by mapping the molecular lesions associated with chromosomally rearranged lab alleles. We present its molecular structure, nucleotide sequence, and temporal pattern of expression. In addition, using antibodies generated against a fusion protein, we show that in the embryo the lab protein is distributed in neural and epidermal cells of the procephalic lobe; in a discrete loop of the midgut; and in specific progenitor sensory cells of the clypeolabrum, thoracic segments, and tail region. The regions of lab expression in the developing cephalon represent nonsegmented domains that are anterior to and largely nonoverlapping with the domains of expression of the Deformed (Dfd) and proboscipedia (pb) genes, two other homeotic loci of the ANT-C that also function to direct the development of head structures. Furthermore, lab head expression is associated with the complex cellular movements of head involution, a process that not only is defective in lab embryos, but the failure of which appears to be largely responsible for the defects observed in mutant embryos. Finally, we suggest that lab head expression provides a molecular marker for an intercalary segment, an ancestral segment that has become morphologically indistinct during the evolution of the insect head. PMID- 2566559 TI - Progressively restricted expression of a homeo box gene within the aboral ectoderm of developing sea urchin embryos. AB - A homeo box-containing gene, Hbox1 is expressed in an unusual and highly conserved spatial pattern in embryos of two different species of sea urchin, Tripneustes gratilla and Strongylocentrotus purpuratus. Hybridization in situ shows that this mRNA accumulates initially throughout the aboral ectoderm; however, between blastula and pluteus stages, the region containing Hbox1 mRNA retracts gradually until only a small area around the vertex is labeled in pluteus larvae. Aboral ectoderm appears cytologically uniform and also accumulates uniform levels of other tissue-specific mRNAs. Therefore, the Hbox1 pattern reveals a previously unsuspected heterogeneity of aboral ectoderm cells and a polarity within this tissue. In S. purpuratus, the Hbox1 gene product probably is not involved in initial specification of cell fate, as this message does not achieve a significant fraction of its peak abundance until almost hatching blastula stage, well after the time aboral ectoderm cells have initiated a tissue-specific program of gene expression. RNA blot and RNase protection analyses revealed low levels of Hbox1 mRNA in all adult tissues examined. However, this message was not detectable in mature eggs, suggesting that the Hbox1 gene does not have a maternal function. In addition to highly conserved spatial and temporal patterns of expression, the homeo box genes of these two urchin species also are conserved highly in sequences outside the homeo domain, despite the divergence of these two species (30-45 my). Two notable features of the protein shared with several vertebrate homeo proteins are a short conserved sequence encoded by an exon upstream of that encoding the homeo domain and a large region of high serine and proline content. PMID- 2566561 TI - [Improving the organizational aspects of coordinating research on the problem of "scientific principles of environmental hygiene"]. AB - A retrospective and scientific metric analysis of the work of the task-force committee "Scientific principles of environmental hygiene", its sections and subordinate institutions was carried out. The analysis demonstrated a major role of hygienic institutes in the development of the above problem and the least contribution of research epidemiologic, microbiologic and hygienic institutes along with chairs of medical institutes. Emphasis was given to a low share (33%) of materials introduced at the All-Union level in relation to the whole amount of the research on the problem, to easing of control on the realization of plans for introducing research results into practice since 1981 by the sections of the task force committee, inconsistence of the research subjects and their results introduced. Attention was paid to the shortcomings in the work of the task-force committee and its sections, necessity of their work intensification, primarily in planning which is one of major levels in raising efficacy of the research, their orientation at solving priority problems. PMID- 2566563 TI - [Pain measurement and a multi-stage plan in gynecologic oncology]. PMID- 2566562 TI - Disabling anxiety. PMID- 2566564 TI - [Treatment of tumor pain]. PMID- 2566565 TI - [Electroencephalographic study on the central action of a new anxiolytic: 3a alpha, 4 beta, 7 beta, 7a alpha-hexahydro-2-(4-(4-(2-pyrimidinyl)-1- piperazinyl) butyl)-4, 7-methano-1H-isoindole-1, 3(2H)-dione dihydrogen citrate (SM-3997)]. AB - Electroencephalographic (EEG) studies were performed to examine the effects of SM 3997 on the spontaneous EEG, EEG arousal responses, recruiting responses and hippocampal afterdischarges in rabbits and the spontaneous EEG in chronically electrode-implanted rats. In acute experiments using rabbits, SM-3997 at doses of 1-3 mg/kg, i.v., produced low-voltage fast waves in cortical EEG and slow waves with reduction of the amplitude in hippocampal EEG. The drug at doses of 1-3 mg/kg, i.v., dose-dependently inhibited the threshold stimulus voltages in EEG arousal responses induced by stimulation of the midbrain reticular formation and slightly inhibited the threshold in recruiting responses by stimulation of the centromedian nucleus of the thalamus. However, the cortical and hippocampal afterdischarges induced by hippocampal stimulation remained unaffected by SM-3997 at doses up to 3 mg/kg, i.v., while they were inhibited by diazepam of 1 mg/kg, i.v. In the study using rats in which electrodes were chronically implanted, SM 3997 at doses of 10-30 mg/kg, i.p., also produced low voltage fast waves in cortical EEG and slow waves of reduced amplitude in hippocampal EEG; and it simultaneously caused flat body posture. These results suggest that SM-3997 acts on both the cerebral cortex and hippocampus, inducing much more pronounced inhibition on the midbrain reticular formation-hippocampal system PMID- 2566566 TI - [Norrie syndrome: identification of carriers by segregation analysis with flanking DNA markers]. AB - Norrie disease is an X-linked recessive disorder. Affected males present with congenital blindness. Additionally, hearing loss and psychotic behavior may occur at any time. Since carriers are clinically healthy, they can only be identified by genetic means. Daughters of carriers or sisters of affected males have an a priori 50% risk of being carriers themselves. Close linkage has been found between the Norrie disease locus (NDP) and the DNA locus DXS7 mapped to Xp11.3. For genetic counselling, this linkage relationship allows carriers of the disease to be identified in informative families. We describe a large pedigree with Norrie disease. Segregation analysis was carried out with DXS7 and a second flanking marker, DXS255, both linked to NDP. In this way, three females at risk were identified who had a high probability of being carriers for Norrie disease. PMID- 2566568 TI - [Allergy treatment--10 years' results using astemizole. Proceedings of the International Hismanal Symposium. Garmisch-Partenkirchen, 24-26 February 1989. Abstracts]. PMID- 2566567 TI - [Staged therapy of chronic obstructive respiratory tract diseases. Treatment of chronic obstructive respiratory tract diseases by the established physician]. PMID- 2566569 TI - [Genetic analysis of familial polyposis coli. Linkage analysis and loss of heterozygosity in colorectal tumors of FPC by RFLP analysis]. AB - Familial polyposis coli (FPC) is a genetic disorder, transmitted as an autosomal dominant trait, characterized by numerous colorectal adenomas. If untreated, most patients may develop colorectal adenocarcinomas. We investigated linkage between FPC and several DNA markers, and loss of heterozygosity in colorectal tumors by using RFLP analysis. We examined 15 pedigrees for linkage analysis and 31 FPC patients including 16 adenocarcinomas and 43 adenomas and 15 non-polyposis colorectal carcinomas (NPCC) for searching loss of heterozygosity. 1. Significant linkage was not observed with 26 polymorphic DNA probes. Maximum lod score of 0.301 at a recombination fraction of 0.0 was observed with the marker D5S71, which was reported to be tightly linked to the major gene for FPC in Caucasian. 2. Loss of heterozygosity was observed at the loci on 17 chromosomes in colorectal carcinomas from FPC patients, and on 6 chromosomes in NPCC. Thus, chromosomes in FPC patients may be unstable compared with those in patients with NPCC. 3. Frequent loss of heterozygosity in colorectal carcinomas from FPC patients was observed on chromosomes 5 (20%), 14 (22%), 17 (43%) and 22 (38%), and was also observed on chromosomes 5 (33%), 14 (38%), 17 (27%) and 22 (15%) in NPCC. These results suggest that tumor suppression genes may locate on these chromosomes. 4. Tumor suppression genes may play a role by dose dependent way. PMID- 2566570 TI - In vitro desensitization of beta-adrenoceptors in guinea pig trachea: interactions between beta-adrenoceptor agonists and influence of adenosine and other drugs. AB - The aim of this study was to investigate quantitatively the action of and the interaction between beta-adrenergic receptor agonists in desensitizing guinea pig isolated trachea. It was also to evaluate the influence of substances whose effects on desensitization are either disputed (theophylline, indomethacin, ketotifen, hydrocortisone) or unknown (nicardipine, Bay K 8644, fenspiride, adenosine). Tracheal strips were contracted with histamine (5 x 10(-5) M) or acetylcholine (5.10(-5) M) and concentration-response (C/R) curves for various beta-adrenoceptor agonists were determined before and after incubation (20 min to 4 h) with the same beta-adrenoceptor agonist (autodesensitization), with other beta-adrenoceptor agonists (cross-desensitization), or with a beta-adrenoceptor agonist and another substance. Our results show that the autodesensitization induced by isoprenaline is concentration dependent and that concentration dependence is more pronounced with salbutamol and fenoterol than with isoprenaline and adrenaline with respect to autodesensitization: shifts (log unit) of the C/R curves were 0.59 +/- 0.06 (N = 5) for salbutamol (10(-5) M), 0.78 +/- 0.09 (N = 5) for fenoterol (10(-6) M), 0.30 +/- 0.04 (N = 9) for isoprenaline (10(-5) M), and 0.33 +/- 0.05 (N = 5) for adrenaline (10(-5) M). Our studies of cross-desensitization (desensitization to isoprenaline, adrenaline, salbutamol, and fenoterol induced by incubation with isoprenaline 10(-5) M) showed a significantly greater shift in the C/R curves for fenoterol (0.56 +/- 0.08, N = 5) and salbutamol (0.62 +/- 0.05, N = 5) than for adrenaline (0.35 +/- 0.07, N = 5) and isoprenaline itself (0.30 +/- 0.05, N = 9). Of the substances we studied, none modified the desensitization induced by isoprenaline except hydrocortisone and adenosine. Hydrocortisone (10(-8) M) reduced it significantly, although to a negligible extent. Adenosine (3 x 10(-4) M) did not shift the C/R curve to isoprenaline by itself, but incubation of tracheal strips with adenosine and isoprenaline caused a significantly greater shift of C/R curves to isoprenaline (0.30 +/- 0.04) than incubation with isoprenaline alone (0.20 +/- 0.04) (P less than 0.05, N = 5). These experiments suggest that adenosine may have increased the uncoupling and/or down-regulation phenomena induced by isoprenaline, or modified adenylate cyclase-cAMP activity. PMID- 2566571 TI - Gastrin and somatostatin levels in patients with gastric cancer. AB - Gastrin and somatostatin-like immunoreactivity (SLI) levels were studied by means of radioimmunoassay in peripheral venous blood of healthy volunteers and patients suffering from gastric adenocarcinoma or duodenal and gastric ulcers. Gastrin and SLI levels were also evaluated in patients in blood drawn from gastric veins during surgery. The elevations of gastrin and SLI levels were found in patients with gastric cancer as compared with healthy people and patients suffering from ulcers. The impairment of the negative feedback between gastrin and somatostatin secretions in patients with gastric cancer was suggested. PMID- 2566572 TI - Receptors and neurohormones in human pituitary adenomas. AB - In order to go further into the pathogenesis of human pituitary adenomas, we studied receptors for neurohormones (thyroliberin, TRH; dopamine, DA; somatostatin, SRIH), for estradiol and epidermal growth factor (EGF) thought to influence hormone secretion and/or cell growth. The following results were obtained: (1) the receptors listed above, with the exception of EGF receptors in the adenomas, are present in normal pituitary tissue and in prolactin (PRL)- and growth hormone (GH)-secreting adenomas; (2) they are functional and their affinities are not different in normal or tumoral tissues; (3) their density is variable and depends on the type of secreting adenoma (GH or PRL), the size of the tumor and the plasma level of the hormone which is secreted, and (4) in nonsecreting adenomas, only TRH receptors are found with characteristics identical to those observed in secreting adenomas. We also showed that TRH is contained in normal and tumoral pituitary tissues. TRH and SRIH are released in vitro from adenomatous cells in large amounts, suggesting their possible synthesis by the pituitary. In both cases a local regulation is observed. TRH release is stimulated in the presence of DA while SRIH is inhibited in the presence of TRH. This neuropeptide release may be implicated in the pituitary hormone regulation through a paracrine or an autocrine mechanism. Thus, the neurohormone receptors found in pituitary adenomas should be dependent on a more complex regulation than it has been envisaged till now. PMID- 2566573 TI - Somatostatin receptors in brain and pituitary. AB - Somatostatin (SRIF) actions in the brain and pituitary are mediated by specific receptors. Using radioiodinated ligands it has been possible to characterize the kinetics of specific binding sites in the brain and pituitary, and to determine their cellular localization by autoradiography. At the pituitary level, the inhibition of growth hormone, prolactin and thyrotropin secretions induced by SRIF is mediated through a single binding site which is coupled to the inhibition of adenylate cyclase. In the brain, SRIF receptors are localized on neurons and glial cells and are also coupled to adenylate cyclase inhibition. Two sites are differentiated in the brain with an analogue of somatostatin, SMS 201995. In humans, SRIF-binding sites have been related to a number of pathologies. At the pituitary level, it has been shown that the number of binding sites was negatively correlated to growth hormone levels in acromegaly. Furthermore, SRIF binding sites were undetectable in a patient which did not respond to SMS 201995 therapy. In the brain, meningiomas and gliomas are rich in SRIF binding sites. This suggests a possible role for SRIF on glia. In neurodegenerative diseases, cortical SRIF concentrations are decreased in Alzheimer's and Parkinson's disease associated with dementia while SRIF-binding sites are only affected in Alzheimer's disease. In conclusion, the physiological role of SRIF in the brain and pituitary can be evaluated by studying the receptors of the peptide. Such studies allow to question the implication of SRIF in endocrine and neuropathologies. PMID- 2566574 TI - Multiple transduction mechanisms of dopamine, somatostatin and angiotensin II receptors in anterior pituitary cells. AB - The concept of multifactorial pituitary control is now well established. As in other cell systems, integration of complex messages involves dynamic interactions of receptors and coupling mechanisms. Regulation of adenohypophyseal secretions has been shown to involve cyclic AMP production, the modulation of phosphatidylinositol phosphate breakdown and Ca2+ mobilization. Dopamine, somatostatin and angiotensin II receptors are negatively coupled to adenylate cyclase in anterior pituitary cells. In the case of angiotensin, this effect on adenylate cyclase appears paradoxical since the peptide markedly stimulates prolactin secretion. In fact, angiotensin II also markedly stimulates inositol phosphate production and this effect could account for the stimulated hormone secretion. In addition, dopamine could inhibit inositol phosphate production stimulated by angiotensin II and thyrotropin-releasing hormone. Dopamine and somatostatin also directly modulate voltage-dependent calcium channels, perhaps through a direct coupling with potassium channels. On the other hand, steroids modulate the sensitivity of adenohypophyseal cells to neurohormones by different mechanisms. In the case of somatostatin, it increases the number of specific binding sites, while in the case of dopamine estradiol affects the transduction mechanisms of D2 dopamine receptors. In conclusion, dopamine and somatostatin receptors appear coupled to various transduction mechanisms through pertussis sensitive G proteins in anterior pituitary cells. PMID- 2566575 TI - Family and population studies on the human pepsinogen A multigene family. AB - Human pepsinogen A (PGA) displays highly polymorphic isozymogen patterns after polyacrylamide gel electrophoresis and activity staining. The patterns differ with respect to the presence and the relative intensity of the individual fractions. Family studies strongly suggest that these isozymogen patterns are encoded by allelic haplotypes, encompassing different numbers and types of PGA genes. In this paper, we confirm the essential features of this multigene model. We establish the relationship between the haplotypes and the corresponding isozymogen patterns by determination of the PGA polymorphism at both the DNA and the protein level in 117 Dutch individuals, 60 of whom were unrelated. The combination of HindIII and EcoRI restriction fragment length polymorphisms (RFLPs) has enabled us to define different haplotypes, which are shown to segregate within families. Most genes are characterized by their specific EcoRI fragments. The HindIII RFLP is in strong linkage disequilibrium with PGA genes showing strong expression of the relevant isozymogen. Although a general picture of the relationship between genotypes and phenotypes is emerging, there are exceptions, suggesting that rare haplotypes evolve by unique crossover events. PMID- 2566576 TI - The levels of zeta, gamma, and delta chains in patients with Hb H disease. AB - Details are given of a study of blood samples from 24 patients with Hb H disease from different Mediterranean countries and from the Far East. Four different types of alpha-thal-1 (--) were observed, namely -(alpha) (approximately 20.5-kb deletion); --MED-I (approximately 17.5-kb deletion); --MED-II (greater than 26.5 kb deletion); and --SEA (approximately 18-kb deletion, in Orientals only). The alpha-thal-2 was mainly of the deletion type (16 with the 3.7-kb deletion; 1 with the 4.2-kb deletion), while 4 of the 7 patients with a nondeletional type had the five-nucleotide deletion at the donor splice site of the first intron of the alpha 2 gene. All patients had a mild-to-moderate hemolytic anemia; no significant differences in hematology were observed between the groups. Hb A2 was decreased to about one-third of the normal level. The Hb H formation varied considerably and its quantitation was not always satisfactory. Patients with Hb H disease due to any alpha-thal-1 combined with a nondeletional alpha-thal-2 had the highest Hb H levels and a more marked anemia. The zeta chain production was small and absent in patients with the MED-II type of alpha-thal-1 because this deletion included the zeta and psi zeta genes. The highest zeta chain levels were present in the four patients with the SEA type of alpha-thal-1. The gamma chain production was increased, particularly in patients with a mutation of C----T at position -158 to the G gamma globin gene. This gamma chain was primarily present as Hb Bart's (or gamma 4) and only about 15% was recovered as Hb F or alpha 2 gamma 2. The evaluation of the rate of gamma chains produced in these patients was greatly facilitated by data from one patient who had Hb H disease and a heterozygosity for the A gamma-beta+-HPFH. The low levels of Hb A2 and of Hb F (relative to Hb Bart's) can be explained by a decreased affinity of alpha chains for delta and gamma chains as compared with beta chains in conditions of severe alpha chain deficiency. PMID- 2566577 TI - Evidence that specific dopamine-1 receptor activation is involved in dopamine induced renin release. AB - Direct effects of dopamine on renin release were examined using static incubations and perifusions of rat renal cortical slices. Dopamine (10(-5)M) significantly stimulated renin release compared with control. To determine which receptors are involved in dopamine-elicited renin release, studies were performed with specific dopamine-1 and dopamine-2 receptor agonists and antagonists, as well as with alpha- and beta-adrenergic antagonists. Fenoldopam, a dopamine-1 receptor agonist, dose dependently stimulated renin secretion both in static incubations and perifusions; whereas quinpirole (10(-7)-10(-5)M), a dopamine-2 receptor agonist, was ineffective. Phentolamine (10(-4)M), an alpha-adrenergic antagonist, did not alter dopamine- or fenoldopam-induced renin release. Similarly, propranolol, a beta-blocker, did not interfere with the renin stimulation of dopamine (10(-5)M) or fenoldopam (10(-6)M) incubations or perifusion experiments; whereas propranolol significantly blocked isoproterenol action. SCH 23390 (10(-5)M), a specific dopamine-1 antagonist, blocked dopamine- and fenoldopam-induced renin. In contrast, pimozide, a dopamine-2 receptor antagonist, was ineffective. These studies indicate that dopamine is a direct renin secretogogue, and its effects seem to be mediated by specific dopamine-1 receptor activation, as neither alpha- nor beta-adrenergic blockers nor dopamine 2 receptor antagonists altered dopamine actions. The results suggest that dopamine produced locally in the kidney may stimulate renin secretion directly by dopamine-1 receptor activation. PMID- 2566578 TI - An increased pool of secretory hormones and peptides in adrenal medulla of stroke prone spontaneously hypertensive rats. AB - Secretory components of the adrenal medulla were compared in normotensive Wistar Kyoto (WKY) rats and in stroke-prone spontaneously hypertensive rats (SHRSP) at both 4 and 12 months of age. Noradrenaline, adrenaline, dopamine, neuropeptide Y, and chromogranins A and B were significantly higher in adrenal glands of SHRSP than those of WKY rats at 4 months. At 12 months, the levels of these components in SHRSP had increased even more (about 200% in WKY rats). There was no change in the relative composition of the adrenal "secretory cocktail." Neither the chromogranin A/chromogranin B ratio nor their apparent proteolytic processing in chromaffin granules differed between SHRSP or WKY rats. The lack of a significant change in membrane-bound cytochrome b561 and the small increase in dopamine beta hydroxylase suggest that the higher levels of secretory components in SHRSP are not simply caused by an increase in the number of chromaffin granules, but possibly by a selective increase in the secretory content of these organelles providing a larger package for quantal release by exocytosis. This may be relevant for the elevation of blood pressure in this strain. The immunological methods described in this paper allow for the first time a determination of the secretory quantal levels in catecholamine storage. This should be useful for further studies in hypertensive models. PMID- 2566579 TI - Evidence that glutamate mediates axon-to-Schwann cell signaling in the squid. AB - High-frequency stimulation (100 Hz) of isolated giant axons of the small squid Alloteuthis subulata and the large squid Loligo forbesi caused the periaxonal Schwann cell resting potential (Em = -40 mV) to hyperpolarize up to 11 mV in direct proportion to train duration and action potential amplitude. In both species, the Schwann cell also hyperpolarized up to 17 mV with the application of L-glutamate (10(-9) to 10(-6) M), in a dose-dependent manner. By contrast, in the presence of 10(-8) M d-tubocurarine (d-TC) to block the cholinergic component of the Schwann cell response, Schwann cells depolarized 8-9 mV during electrical stimulation of the axon or application of L-glutamate. In the presence of 10(-5) M 2-amino-4-phosphonobutyrate (2-APB), the hyperpolarization to glutamate and to axon stimulation was blocked, whereas the cholinergic (carbachol-induced) hyperpolarization was unaffected. In experiments with Alloteuthis, L-aspartate (10(-7) M) also caused a Schwann cell hyperpolarization, but this was not blocked by 2-APB. In tests with glutamate receptor agonists and antagonists, quisqualate (10(-5) M) produced a hyperpolarization blocked by 10(-4) M L-glutamic acid diethylester (GDEE), which also blocked the response to axonal stimulation. Kainic acid (10(-4) M) also caused a hyperpolarization, but n-methyl-D-aspartate (NMDA; 10(-4) M), ibotenate (10(-5) M), alpha-amino-3-hydroxy-5-methyl-isoxazole proprionate (AMPA; (10(-4) M), and isethionate (10(-5) M) had no effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566580 TI - In vivo expression and variation of Escherichia coli type 1 and P pili in the urine of adults with acute urinary tract infections. AB - In vivo expression of pili by Escherichia coli in the urine of 41 adults with lower urinary tract infections was analyzed by immunostaining with polyclonal antiserum to type 1 and P pili. Type 1 pili were detected in 31 of 41 urine specimens, while P pili were detected in 6 of 18 specimens. The piliation status of bacterial populations in urine was heterogeneous, varying from predominantly piliated to a mixture of piliated and nonpiliated cells. Bacteria frequently adhered to exfoliated uroepithelial cells and leukocytes in urine. Expression of pili in vivo did not always correlate with the hemagglutination phenotype after growth in vitro. Strains isolated from different sites in the urogenital tract of two individuals showed phenotypic variation in the state of piliation. The results demonstrate that E. coli type 1 and P pili are expressed and are subject to variation in vivo during acute urinary tract infections in adults. PMID- 2566582 TI - Anesthesia-induced dental injury. PMID- 2566581 TI - Cloning and temperature-dependent expression in Escherichia coli of a Legionella pneumophila gene coding for a genus-common 60-kilodalton antigen. AB - All Legionella species express a 60-kilodalton (kDa) protein which contains a genus-specific epitope recognized by murine monoclonal antibody GW2X4B8B2H6. A genomic cosmid library of Legionella pneumophila chromosomal DNA was constructed in pHC79 and screened for 60-kDa antigen-expressing clones with the monoclonal antibody. A 3.2-kilobase EcoRI fragment from cosmid 14B11 expressing a 60-kDa protein was subcloned into pUC19 (pSH16), and deletion of a 1.2-kilobase HindIII fragment (pSH16A) generated a 33-kDa truncated polypeptide no longer reactive with the monoclonal antibody. Southern blot analysis of chromosomal DNA from selected Legionella species restricted with EcoRI and probed with the 1.2 kilobase fragment coding for the carboxyl region of the protein revealed DNA homology which was not observed with DNA from Escherichia coli. Maxicell analysis of pSH16 identified a second polypeptide of approximately 15 kDa expressed from a gene (htpA) upstream of the gene coding the 60-kDa protein (htpB). Both proteins were preferentially synthesized by L. pneumophila following heat shock (temperature shift from 25 to 42 degrees C), and under steady-state growth conditions the relative level of 60-kDa protein was unaffected by temperature. In E. coli, expression of a 60-kDa protein from pSH16 also increased following heat shock (25 to 42 degrees C), but under steady-state conditions expression was temperature dependent. Temperature-dependent expression from pSH16 was not observed in an rpoH (htpR) mutant strain of E. coli. The Legionella 60-kDa protein appears to be a heat shock protein which shares cross-reactive epitopes with the GroEL homolog of E. coli. In addition, a region of htpB encoding the 27 kDa carboxyl portion of the protein containing the monoclonal antibody-reactive epitope also contains DNA sequences unique to and conserved within the genus. PMID- 2566583 TI - Intravenous sedation in dentistry and oral surgery. AB - IV administration is the most precise and effective means of sedating a patient. Its desired effect is a comfortable and cooperative patient whose pain is alleviated by regional local anesthesia. It requires a patient who understands the difference between being lightly sedated and unconscious. Its primary purpose is to diminish anxiety and apprehension rather than to obtund protective reflexes. At all times, the patient should remain conscious and appropriately responsive to questions or commands. This state can be readily achieved in most patients by carefully titrating a single drug such as diazepam or midazolam to effect. The use of multiple drugs is to be discouraged, since it generally increases the level of sedation and the number of complications. Dosing should be individualized rather than averaged or arbitrary. The end point of titration is the patient's verbal acknowledgement of feeling more relaxed and the physical evidence of such relaxation. The "Verrill sign" indicates a level of deep sedation--too deep for most cases of conscious sedation in the office setting. I strongly oppose the routine use of narcotics for office sedation. Monitoring of vital signs before, during, and after surgery ensures the safety of the consciously sedated patient. The use of a pulse oximeter is an evolving standard in general anesthesia and may eventually prove important for patients receiving conscious sedation in the office setting. PMID- 2566584 TI - Proposed role of calcium in reperfusion injury. PMID- 2566585 TI - Esmolol (Brevibloc) to assess dynamic subpulmonary stenosis in a child after repair of complete transposition: a case report. AB - Esmolol (Brevibloc), a new, ultra-short acting, cardioselective beta-adrenergic blocking agent with half-life of 9.2 min following i.v. administration was given to a 4-year-old child with known dynamic and fixed sub-pulmonary stenosis post Senning repair for complete transposition of the great vessels. The left ventricular systolic pressure increased from 48 to 100 mmHg, heart rate showed an increase from 65 to 140 bpm, the right femoral arterial pressures decreased from 115/58 to 77/35 mmHg following an infusion of Isoprel. Infusion of esmolol partially relieved the dynamic sub-pulmonary stenosis. There were no adverse effects and esmolol was tolerated well by the child. Esmolol might thus play a role in the pediatric catheterization laboratory during investigational procedures, electrophysiological studies and in the control of rapid supraventricular tachycardia, especially in adolescents with WPW pre-excitation. Esmolol would also be beneficial in emergency treatment of epinephrine or isoproterenol overdosage. PMID- 2566586 TI - Presynaptic modulation of sympathetic nerve transmission--an element in vasomotor control. AB - We have earlier presented evidence that adenosine may create vasodilation via presynaptic inhibition of sympathetic nerve fibers to muscle arterioles. In the present study the effect of selective adenosine antagonists on arteriolar diameter was investigated, using an autoperfused frog muscle (m. cutaneus pectoris). During application of the antagonists arteriolar diameter reductions were observed in the transparent, transilluminated muscle. The sulfonylated xanthine derivatives 8-(p-sulfo)phenyltheophylline (8-PS phi T), 8-(m sulfo)phenyltheophylline (8-MS phi T) and 1,3-dipropyl-8-p-sulfonphenylxanthine (DPSPX) are all considered to be selective competitive adenosine antagonists without unspecific intracellular effects. Topical application of each of these compouns on resting muscle resulted in a dose-dependent arteriolar contraction. The constriction was immediately reversed by addition of the adrenergic alpha receptor blocker phentolamine (7.1 x 10(-5) M), indicating that the arteriolar contraction was due to activation of postjunctional alpha-receptors present in vascular smooth muscle. Also, the A1-selective adenosine agonist (-)-N6-(R phenylisopropyl)-adenosine (R-PIA) eliminated the arteriolar constriction. These effects suggest, that the xanthine derivatives release the sympathetic nerve terminals from inhibition by endogenous adenosine. When nerve fibers were blocked by tetrodotoxin (TTX), 3 x 10(-6)M or when the frogs had been chemically sympathectomized with 6-hydroxydopamine (6-OHDA), no significant arteriolar contraction was observed during adenosine antagonist administration, complying with the view that in resting muscle endogenous adenosine acts presynaptically to inhibit sympathetic nerve terminals. These results suggest that presynaptic adenosine effects are not only involved in regulation of frog skeletal muscle blood flow in response to activity, but also play a role in the control of arteriolar tone in resting muscle. PMID- 2566587 TI - Neurophysiology and neurochemistry of drug dependence: a review. AB - To understand the neurophysiological and neurochemical mechanisms of drug dependence, the functional significance of dopamine, noradrenaline and endogenous opioid peptides in the mediation of natural, self-stimulation and pharmacological reinforcement are discussed. Data on search of system(s), mediator(s) and neurons of reinforcement as well as my own notions on reinforcement as a critical element in organization and regulation of the organism's adaptive activity in variable environments are presented. The role of chronic drug-induced stable modification of central neurochemical systems' functioning as a basis for the alteration of endogenous reinforcement processes and raising drug dependence are examined in detail for main addictive drugs, opiates and psychomotor stimulants. PMID- 2566588 TI - Characterization of southern African isolates of maize streak virus: typing of three isolates by restriction mapping. AB - The genomic replicative form DNAs (RF-DNA) of three maize streak virus isolates (MSV-CT, MSV-PE, and MSV-SW) from widely separated locations in southern Africa were characterized by restriction endonuclease mapping in order to assess the feasibility of using the technique to determine genetic variability between isolates. The viruses were transmitted to and propagated in laboratory-grown maize by the leafhopper vector Cicadulina mbila (Naude). MSV-PE produced more severe symptoms than MSV-CT and MSV-SW; the isolates were serologically identical in 'western' immunoblot tests, but distinct in 'sandwich' enzyme-linked immunosorbent assays. RF-DNA of all three isolates was prepared from infected maize; the RF-DNA of MSV-CT and MSV-PE was cloned in a plasmid vector in Escherichia coli. Restriction maps were generated from this cloned DNA and from the RF-DNA of MSV-SWA. The maps were similar in regions expected to be conserved, but there were also important differences between all isolates. The implications of these results, and of relationships amongst these and other sequenced isolates of MSV, are discussed. PMID- 2566589 TI - Microgeographic variation in rDNA intergenic spacers of Anopheles gambiae in western Kenya. AB - The genetic population structure of Anopheles gambiae (Diptera: Culicidae) in western Kenya was investigated by hybridizing a rapidly evolving rDNA intergenic spacer sequence to restriction endonuclease digests of genomic DNA extracted from single mosquitoes from seven localities. Significantly different distributions of restriction fragment arrays were obtained from field sites less than 10 km apart, which suggests restricted gene flow and a subdivided population structure. Eight of twenty-one possible comparisons between pairs of populations yielded significant differences. An eastern Kenya coastal population did not share its restriction fragment arrays with any of the western populations, suggesting that isolation by distance can be complete on a relatively small geographic scale (700 km). PMID- 2566590 TI - Thermotolerance in the mouse foot estimated at various levels of normal tissue damage. AB - The effect of fractionated 43.7 degrees C water bath heating on the skin of CDF1 mice was investigated. The normal tissue damage was scored at five levels (from slight redness and oedema to loss of a toe or greater damage) according to an arbitrary score system. The heating time to induce a given level of damage in half of the treated animals (RD50) was used as an end point. The feet were exposed either to a single treatment at 43.7 degrees C for different time periods or to a priming treatment of 30 min. at 43.7 degrees C followed at different intervals by a second graded heat treatment at 43.7 degrees C. In all treatment schedules, the score level increased proportionally with heating time, and the score system offered a good description of the acute skin damage following hyperthermia. The priming heat treatment induced thermotolerance with a time course independent of the score level chosen to estimate the heat response. The thermotolerance developed rapidly, reached a maximum within a 24 hr. interval, and then decayed slowly. The degree of thermotolerance was calculated by means of two previously described formulas for the thermotolerance ratio (TTR). The kinetics of thermotolerance in the skin of mice was independent of the TTR formula, whereas the degree of thermotolerance depended on both the score level and the TTR formula used. PMID- 2566591 TI - The biphasic change of cytosolic acetyl-CoA hydrolase in rat liver during 3' methyl-4-dimethylaminoazobenzene hepatocarcinogenesis. AB - When Donryu male albino rats were given diet containing 0.06% 3'-methyl-4 dimethyl-aminoazobenzene (3'-Me-DAB) for 20 weeks, the activity of cytosolic acetyl-CoA hydrolase in their livers decreased to about one-third the initial level in week 2, returned to the control level in week 7, and then decreased again to about one-tenth of the control in week 20. These changes in enzyme activity were parallel with changes in the amount of enzyme protein determined by ELISA. In 3'-Me-DAB-resistant rats, however, the enzyme activity and enzyme protein remained within the normal range during administration of 3'-Me-DAB containing diet for 20 weeks and no tumors were detectable macroscopically. Interestingly, the biphasic change in this enzyme activity was inversely associated with the well known change of gamma-glutamyltranspeptidase activity during azo-dye-induced hepatocarcinogenesis. PMID- 2566592 TI - Selective killing of human T cell lymphotropic virus type I-transformed cell lines by a damavaricin Fc derivative. AB - n-Pentyl ether of damavaricin Fc (n-pentyl DvFc) preferentially killed human T cell lymphotropic virus type I (HTLV-I)-transformed cell lines. The mechanism of action of the drug was investigated using MT-4 cells. Cytotoxic action was diminished by the removal of n-pentyl DvFc from the culture or by the addition of sulfhydryl compounds such as 2-mercaptoethanol and dithiothreitol. The killing activity of n-pentyl DvFc was also diminished by membrane-acting agents including quinidine and diphenylhydantoin. Influx and subsequent efflux of Ca2+ were observed when either HTLV-I infected (MT-4 cells) or uninfected cells were treated with n-pentyl DvFc. An efflux of K+ was observed in HTLV-I infected MT-4 cells immediately after the exposure of the cells to n-pentyl DvFc. The K+ efflux, however, was not observed in the uninfected T cells. n-Pentyl DvFc seems to act primarily on the cell surface of MT-4 cells, leading to the perturbation of membrane function. The restoration of cell growth, however, is critically dependent on the presence of dithiothreitol and 2-mercaptoethanol, implying a role for a free sulfhydryl group in the killing activity. PMID- 2566593 TI - Effects of cimaterol on nitrogen retention and energy utilization in lambs. AB - Fifty-two weaned lambs were used in a comparative slaughter feeding trial and six lambs in a concurrent digestibility trial to investigate the effects of cimaterol on nitrogen retention and energy utilization. Cimaterol (CIM) was administered in the feed at 10 ppm for 90 and 14 d, respectively, in a comparative slaughter feeding trial and in a digestibility trial. No difference was found in dry matter digestibility. Nitrogen retention in the CIM group (739 mg/Wkg.75/d) was greater (P less than .01) than that of the control group (321 mg/Wkg.75/d). This difference was accounted for primarily by reduced nitrogen loss in urine of the CIM group. Cimaterol improved growth rate and feed/gain ratio, although these improvements were only evident during the first 42 d of the 90-d feeding trial. The improvement in growth performance of the CIM group was associated with increased protein gain in the empty body (40.2 g/d, P less than .01) as well as in carcass (26.9 g/d, P less than .01), compared with 30.1 and 14.5 g/d of the control group. Cimaterol decreased (P less than .01) fat gain (91.4 vs 109.9 g/d), total daily energy gain (1.08 vs 1.22 Mcal) and energy gain/kg gain (4.17 vs 5.11 Mcal) compared with the control. Feeding CIM increased (P less than .01) estimated fasting heat production (73 vs 64 Kcal/Wkg.75) and metabolizable energy requirement for maintenance (110.2 vs 92.7 Kcal/Wkg.75) over the control.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566594 TI - Purification and properties of a 28-kilodalton hemolytic and mosquitocidal protein toxin of Bacillus thuringiensis subsp. darmstadiensis 73-E10-2. AB - The mosquitocidal crystal of Bacillus thuringiensis subsp. darmstadiensis 73-E10 2 was purified, bioassayed against third-instar Aedes aegypti larvae (50% lethal concentration, 7.5 micrograms/ml), and subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis, revealing polypeptides of 125, 50, 47, and 28 kilodaltons (kDa). When solubilized and proteolytically activated by insect gut proteases or proteinase K, the crystal was cytotoxic to insect and mammalian cells in vitro and was hemolytic. By using nondenaturing polyacrylamide gel electrophoresis, a polypeptide of 23 kDa, derived from the 28-kDa protoxin, was identified which was hemolytic and cytotoxic to Aedes albopictus, A. aegypti, and Choristoneura fumiferana CF1 insect cell lines. The 23-kDa polypeptide was purified by ion-exchange chromatography and gave 50% lethal dose values of 3.8, 3.3, and 6.9 micrograms/ml against A. albopictus, A. aegypti, and C. fumiferana CF1 cells lines, respectively. Cytotoxicity in vitro was both dose and temperature dependent, with a sigmoidal dose-response curve. The cytotoxicity of the 23-kDa toxin and the solubilized and proteolytically activated delta endotoxin was inhibited by a range of phospholipids containing unsaturated fatty acids and by triglyceride and diglyceride dispersions. An interaction with membrane phospholipids appears important for toxicity. Polyclonal antisera prepared against the 23-kDa polypeptide did not cross-react with polypeptides in the native crystals of four other mosquitocidal strains. PMID- 2566595 TI - Bacteriophage association of streptococcal pyrogenic exotoxin type C. AB - A gene encoding streptococcal pyrogenic exotoxin type C (SPE C) was isolated from bacteriophage DNA derived from Streptococcus pyogenes CS112. The gene, designated speC2, was shown to reside near the phage attachment site of phage CS112. A restriction endonuclease map of the CS112 phage was generated, and the location and orientation of the speC2 gene were determined. Hybridization analyses of eight SPE C-producing strains revealed restriction fragment length polymorphism of the speC gene-containing DNA fragments and further showed that each speC was linked to a common CS112 phage-derived DNA fragment. PMID- 2566596 TI - Glutamine auxotrophs of Bacillus subtilis that overproduce glutamine synthetase antigen have altered conserved amino acids in or near the active site. AB - A number of mutations within the Bacillus subtilis glutamine synthetase (GS) gene result in altered catalytic properties and overproduction of the GS antigen. The restriction fragments containing mutations from three such mutants were sequenced, and they all had amino acid changes in conserved residues found either within or near sequences contributing to the active site of the Salmonella typhimurium GS. PMID- 2566597 TI - Heterogeneity of the gamma-globin gene sequences in Japanese individuals: implication of gene conversion in generation of polymorphisms. AB - The linked fetal globin genes (the G gamma- and A gamma-globin genes) were cloned from Japanese individuals with three different haplotypes of the HindIII polymorphisms within the gamma-globin genes. Determination of nucleotide sequences of the segment spanning from IVS2 to the 3' flanking region of each gamma-globin gene revealed that nucleotide differences are located at 43 positions and a stretch of simple GT or GC sequences. Almost half of the nucleotide changes could be accounted for by gene conversion between the G gamma- and A gamma-globin genes. We found that gene conversion had created the SacI polymorphic site just downstream of the A gamma-globin coding region. Association of the SacI polymorphic site with the HindIII polymorphic site suggests that the region containing these two sites was derived from that of the linked G gamma globin gene through a gene conversion event. The nucleotide sequences obtained here are identical to those of the Caucasoid fetal globin genes of the same haplotypes, with the exception of some sequence changes in the hot spots of mutations. These results indicate that the sequence heterogeneity of the gamma globin genes can be classified into three major categories according to HindIII haplotypes. The possible mechanisms of generation of the heterogeneity of the gamma-globin gene sequences are discussed. PMID- 2566598 TI - Dissociation and auto-oxidation of hemerythrin induced by SH-modification: a kinetic study. AB - Hemerythrin from Siphonosoma cumanense has a trimeric structure consisting of identical subunits, which have no cooperativity nor Bohr effect on oxygen binding. The trimer was dissociated into its monomers by the modification of the SH group of its cysteines with p-chloromercuriphenylsulfonic acid (PCMPS), which was monitored by stopped-flow of both spectrophotomeric and small angle X-ray scattering methods. The results showed that the process involved sequential modification of the SH groups, dissociation into monomers, and auto-oxidation of ferrous iron in the active center. The modification of the SH groups with PCMPS followed second-order kinetics with a rate constant of 1.8 M-1.s-1. The dissociation and auto-oxidation followed first-order kinetics with rate constants of 4 X 10(-3) s-1 and 5 X 10(-4) s-1, respectively. The obtained rate of auto oxidation was much faster than that in the native state. These findings lead to the conclusion that the trimeric state of S. cumanense hemerythrin is necessary to prevent auto-oxidation. PMID- 2566599 TI - 19F-NMR study on the interaction of fluorobenzoate with porcine kidney D-amino acid oxidase. AB - The interactions of competitive inhibitors, o-, m-, and p-fluorobenzoates, with porcine kidney D-amino acid oxidase (DAO) were studied by 19F-NMR spectroscopy. The 19F-signals of DAO-bound fluorobenzoates were observed as considerably broadened peaks. The chemical shifts, which are referenced to 20 mM NaF in 50 mM sodium phosphate, pH 7.0, were 6.0, 8.2, and 11.9 ppm for free o-, m-, and p fluorobenzoates, respectively, while those of o-, m-, and p-fluorobenzoates bound to DAO were 12.5, 5.4, and 13.1 ppm, respectively. The 19F-signals of bound o- and p-fluorobenzoates were downfield-shifted relative to those of the free species, whereas the 19F-resonance of m-fluorobenzoate was up-field shifted from that of the free ligand. The magnitude of the chemical shift difference between the free and bound forms decreases in the order of o-, m-, and p-fluorobenzoates. The remarkably large downfield shift of the o-fluorobenzoate when bound to DAO was attributed to the close proximity of the ortho-fluorine atom to the flavin nucleus in comparison with meta- or para-fluorine. The pH-dependences of the 19F resonances of o-, m-, and p-fluorobenzoates were observed and the pKa values of 3.33, 3.80, and 4.05 were obtained for the carboxyl groups of o-, m-, and p fluorobenzoates, respectively. It was observed that the 19F-resonances of o- and p-fluorobenzoates are highly sensitive to the ionic state of the carboxyl group, while that of m-fluorobenzoate was moderately sensitive. PMID- 2566600 TI - Binding of connectin to myosin filaments. AB - Binding of native connectin (2,100 kDa fragment of alpha-connectin) to myosin filaments was investigated using a sedimentation technique and densitometric estimations of the separated proteins. In the presence of 60 mM KCl and 5 mM phosphate buffer, pH 7.0, as much as 1.5 mol of connectin was bound to 1 mol of myosin, suggesting that some 150 connectin filaments bound to a single myosin filament of approximately 0.5 micron in length. This value was much more than the ratio found in muscle (12:1). It appeared that C protein did not affect the binding of connectin to myosin filaments. PMID- 2566601 TI - In an Escherichia coli coupled transcription-translation system, expression of the osmoregulated gene proU is stimulated at elevated potassium concentrations and by an extract from cells grown at high osmolality. AB - We have studied the in vitro expression of the osmoregulated proU promoter in Escherichia coli coupled transcription-translation (S-30) extracts as a function of the osmolality of the culture medium used to grow the cells and of the salt concentration added to the extract. These variables represent novel extensions of the use of S-30 extracts to investigate gene regulatory phenomena in vitro. The concentrations of potassium acetate and of the physiologically relevant osmolyte potassium glutamate for maximal expression of the proU promoter are approximately 2-fold higher than the concentrations of these salts providing maximal expression of the lacUV5 promoter. The relative promoter activity of proU with respect to lacUV5 increases more than 30-fold with an increase in salt concentration from 50 to 300 mM. In comparative studies with S-30 extracts prepared from cells grown at high and low osmolalities, we find that at fixed salt concentrations expression of proU is increased 10-fold in the S-30 extract prepared from high osmolality grown cells, whereas the expression of lacUV5 is increased less than 2-fold. Addition of anti-sigma 70 monoclonal antibodies or purified sigma 70 to the S-30 extract demonstrated that the major proU promoter(s) used in the S-30 extracts is sigma 70-dependent. PMID- 2566602 TI - Incorporation of monoclonal antibodies into cells by osmotic permeabilization. Effect on cellular metabolism. AB - Incorporation of asparagine synthetase-specific monoclonal antibodies into L5178Y D10/R (L-asparaginase-resistant) murine lymphoma cells by osmotic lysis of pinocytic vesicles was used to evaluate the potential of the technique for macromolecular incorporation for metabolic studies. Nonspecific effects of the incorporation procedure included temporary inhibition of protein and DNA synthesis by 80-85% and a transitory loss of membrane integrity. Cells incorporated with an antibody inhibitory to tumor cell asparagine synthetase showed increased dependence upon an exogenous source of asparagine in the culture medium, while cells incorporated with a control antibody were not affected. These studies demonstrated that incorporation of inhibitory monoclonal antibodies into cells can be used to study the short term metabolic role of specific enzymes; however, the metabolic effects induced by the specific macromolecule must be evaluated within the context of the nonspecific effects caused by the osmotic treatment required for incorporation. PMID- 2566603 TI - Interaction of histidine-rich glycoprotein with human T lymphocytes. AB - Histidine-rich glycoprotein (HRGP), a human plasma and platelet protein, interacts with multiple ligands in vitro, including heparin, plasminogen, thrombospondin, and fibrinogen/fibrin. In this study, the binding of HRGP to human T lymphocytes was characterized. The binding was specific, concentration dependent, saturable, and reversible. Scatchard plot analysis revealed two classes of binding sites: the high affinity class had an apparent dissociation constant (Kd) of 1.92 X 10(-8) M, with 0.92 X 10(4) sites/cell, and the low affinity class had a Kd of 4.97 X 10(-7) M, with 3.7 X 10(4) sites/cell. HRGP binding to T cells in the presence of HRGP-depleted serum was comparable to that observed in buffer. Dot-blot analysis showed that HRGP bound to specific T cell proteins. Using both HRGP affinity chromatography and immunoprecipitation with affinity-purified anti-HRGP IgG, a major 56-kDa HRGP-binding protein in surface labeled T cell lysates was demonstrated. The 56-kDa protein was shown not to be related to the CD2 molecule on T cells. The binding characteristics of HRGP to T lymphocytes indicate a specific ligand-receptor interaction. This is the first demonstration of HRGP binding to a cell surface, and its binding to human T cells may play an important role in T lymphocyte biology. PMID- 2566604 TI - Characterization of a H+-ATPase in rat brain synaptic vesicles. Coupling to L glutamate transport. AB - Synaptic vesicles contain a H+-ATPase that generates a proton electrochemical gradient (delta mu H+) required for the uptake of neurotransmitters into the organelles. In this study, the synaptic vesicle H+-ATPase was examined for structural and functional similarities with other identified ATPases that generate a delta mu H+ across membranes. The synaptic vesicle H+-ATPase displayed immunological similarity with the 115-, 72-, and 39-kDa subunits of a vacuolar type H+-ATPase purified from chromaffin granules. Functionally, the ATP-dependent H+ pumping across synaptic vesicles and ATP hydrolysis were sensitive to the sulfhydryl-modifying reagents, N-ethylmaleimide and 4-chloro-7-nitrobenz-2-oxa 1,3-diazole, at concentrations known to affect vacuolar-type H+-ATPases. In addition, as with vacuolar-type H+-ATPases, the presence of NO3-, SO4(2-), or F- inhibited the generation of a delta mu H+, but addition of vanadate or oligomycin had no effect. The delta mu H+ is a function of the pH gradient (delta pH) and membrane potential (delta psi sv) across the synaptic vesicle. Acidification (delta pH) of the synaptic vesicle interior was enhanced in the presence of permeant anions, such as Cl-, or the K+ ionophore, valinomycin. In the absence of permeant anions, the H+-ATPase generated a delta psi sv that effected the transport of L-glutamate into the synaptic vesicles. Dissipation of delta psi sv by incubation with increased external Cl- or nigericin resulted in the abolition of glutamate uptake, despite the continued maintenance of a delta mu H+ across the synaptic vesicle as a substantial delta pH. The results suggest that the synaptic vesicle H+-ATPase is of a vacuolar type and energizes the uptake of anionic glutamate by virtue of the delta psi sv component of the delta mu H+ it generates. PMID- 2566605 TI - Differential phenotypic expression induced in cultured rat astroblasts by acidic fibroblast growth factor, epidermal growth factor, and thrombin. AB - We compared the effects of three growth factors, acidic fibroblast growth factor (aFGF), epidermal growth factor (EGF), and thrombin, on rat astroblast proliferation, morphology, glutamine synthetase-specific activity, and phenotypic expression of proteins. In vitro experiments were made on 20-day-old primary cultures. Astroblast proliferation was stimulated transiently (after 48 h treatment) by the three growth factors, while the cell glutamine synthetase activity began to increase significantly only after 3 days of treatment. Acidic FGF and EGF, but not thrombin, modified the cell morphology. The effects on phenotypic expression were first determined after 5 days of treatment to minimize the mitogenic effect of the factors. Proteins synthesized during the last 18 h of the treatments were separated by two-dimensional polyacrylamide gel electrophoresis. About 600 spots were compared, 54 were modulated by the various treatments, 13 were altered similarly by all three factors, 28 by aFGF and EGF, 7 by only aFGF, 3 by only EGF, and 3 by only thrombin. These results indicate a large similarity of effects between aFGF and EGF (41 proteins) and show that these factors elicit a more extended modulation of the phenotypic expression than thrombin (13 proteins). Each of the three factors has a few specific effects, which suggests that even for aFGF and EGF, which are supposed to elicit their effects through membrane receptor-associated tyrosine kinase activity, some specificity appears in their mechanism of action. A model is proposed to suggest that cell maturation is characterized by the modulation of the synthesis of many proteins which can be grouped into classes. Each class appears to be under the control of one regulatory element. The specificity of the effect of a growth factor should result from the activation of a specific combination of such regulatory elements. Analysis of the proteins after only 18 h of treatment, when neither proliferation nor maturation were significantly affected, showed that 11 proteins were regulated only at that time. These proteins could be related to intermediate steps of the growth factor signal transduction. PMID- 2566606 TI - Sequential protooncogene expression in regenerating kidney following acute renal injury. AB - Following loss of functional renal mass due to acute injury, there is significantly increased proliferation of tubular epithelium to replace injured and necrotic cells. In contrast, following uninephrectomy, the contralateral kidney increases in size primarily by hypertrophy, with little cellular proliferation. We and others have demonstrated only modest increases in renal protooncogene expression following uninephrectomy. In this study, we demonstrate markedly elevated expression of the protooncogenes c-fos, c-myc, c-Ki-ras, and c Ha-ras following acute renal injury induced by a single large parenteral dose of folic acid. The expression of these genes occurs in a sequential pattern similar to that seen in proliferating cells in culture and in regenerating liver. In addition, we demonstrate elevated levels of histone H4 and beta-actin mRNAs consistent with increased cell proliferation. These data suggest that the molecular mechanisms regulating cell proliferation in the kidney are similar to those in regenerating liver and in cultured cells. In addition, it appears that these events are regulated normally after acute renal injury and following uninephrectomy, since in both instances the levels of protooncogene expression correlate with the degree of cell proliferation. This is in direct contrast to a pathologic renal condition, polycystic kidney disease, in which the level of protooncogene expression is out of proportion to the degree of cell proliferation. Further studies of the molecular correlates of acute renal injury may yield insight into the pathogenesis of this and other clinically important renal disorders. PMID- 2566607 TI - Identification of amino acid residues modified by pyridoxal 5'-phosphate in Escherichia coli glutamine synthetase. AB - Chemical modification studies with pyridoxal 5'-phosphate have indicated that lysine(s) appear to be at or near the active site of Escherichia coli glutamine synthetase (Colanduoni, J., and Villafranca, J. J. (1985) J. Biol. Chem. 260, 15042-15050; Whitley, E. J., Jr., and Ginsburg, A. (1978) J. Biol. Chem. 253, 7017-7025). Enzyme samples were prepared that contained approximately 1, approximately 2, and approximately 3 pyridoxamine 5'-phosphate residues/50,000-Da monomer; the activity of each sample was 100, 25, and 14% of the activity of unmodified enzyme, respectively. Cyanogen bromide cleavage of each enzyme sample was performed, the peptides were separated by high performance liquid chromatography, and the peptides containing pyridoxamine 5'-phosphate were identified by their absorbance at 320 nm. These isolated peptides were analyzed for amino acid composition and sequenced. The N terminus of the protein (a serine residue) was modified by pyridoxal 5'-phosphate at a stoichiometry of approximately 1/50,000 Da and this modified enzyme had full catalytic activity. Beyond a stoichiometry of approximately 1, lysines 383 and 352 reacted with pyridoxal 5'-phosphate and each modification results in a partial loss of activity. When various combinations of substrates and substrate analogs (ADP/Pi or L-methionine-SR-sulfoximine phosphate/ADP) were used to protect the enzyme from modification, Lys-352 was protected from modification indicating that this residue is at the active site. Under all experimental conditions employed, Lys 47, which reacts with the ATP analog 5'-p-fluorosulfonylbenzoyl-adenosine does not react with pyridoxal 5'-phosphate. PMID- 2566608 TI - Dopamine-1-mediated stimulation of phospholipase C activity in rat renal cortical membranes. AB - Phospholipase C (PL-C) mediates transduction of neurotransmitter signals across membranes via hydrolysis of phosphatidylinositol-4,5-bisphosphate (PIP2), leading to generation of second messengers inositol-1,4,5-trisphosphate and diacylglycerol. In this study, dopamine-1 (DA-1) but not dopamine-2 (DA-2) agonists were shown to stimulate PL-C activity in renal cortical membranes. The DA-1 agonist, SKF 82526, stimulated the release of inositol phosphates from renal cortical membranes prelabeled with [3H]myoinositol. The majority of the label (75%) was found in phosphatidylinositol followed by PIP2 (15%) and phosphatidylinositol-4-phosphate (10%). A DA-1 specific effect on PL-C activity was also observed in an in vitro assay of PL-C activity in renal cortical membranes and basolateral and brush border membranes using [3H]PIP2 as the substrate. Dopamine and SKF 82526 stimulated the release of inositol phosphates from added [3H]PIP2 in a concentration-dependent manner. This release was blocked by the DA-1 antagonist SCH 23390 but not by the alpha-adrenergic antagonists phentolamine and prazosin. In contrast, the DA-2 agonist LY 171555 had no effect on inositol phosphate release. Guanosine 5'-(3-O-thio)triphosphate enhanced while guanyl-5'-yl thiophosphate attenuated the DA-1 agonist-stimulated PL-C activity. PL-C activity as measured by [3H]PIP2 hydrolysis had a pH optimum of 6.5, was inhibited by Mg2+ concentrations above 1 mM, was linear with time and protein concentration, and was sensitive to phosphatidylserine and calcium concentrations. We conclude that PL-C is activated by DA-1 but not DA-2 agonists in renal cortical membranes as well as both the basolateral and brush border renal tubular membranes. It is speculated that this action may mediate the natriuretic effects of dopamine in renal tubular epithelia. PMID- 2566609 TI - Factors involved in specific transcription by mammalian RNA polymerase II. Factors IIE and IIF independently interact with RNA polymerase II. AB - The purification and characterization of transcription factor IIF (TFIIF), a factor required for transcription by the RNA polymerase II machinery, is described. TFIIF was isolated from the previously described IIE protein fraction. TFIIF enters into the transcription cycle via a preinitiation complex, and it is required for the formation of a complex capable of initiating transcription in the presence of heparin concentrations that inhibit the action of a free factor. TFIIF and TFIIE independently interacted with purified RNA polymerase II. TFIIF and TFIIE were both required for transcription of several class II promoters, including a promoter that lacks the conserved TATA box. Interestingly TFIIF was absolutely required for the formation of a preinitiation complex; however, it also affected the elongation phase of the transcription cycle. TFIIF, together with the previously described elongation factor TFIIS, was required for efficient elongation. PMID- 2566610 TI - The small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase and its precursor expressed in Escherichia coli are associated with groEL protein. AB - The small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase is synthesized in the cytoplasm as a precursor which is transported into the chloroplast. During or after transport the precursor is processed to its mature size by removal of an amino-terminal transit peptide. Eight small subunits and eight large subunits (synthesized in the chloroplast) assemble to form the holoenzyme. We have expressed the precursor of the small subunit in Escherichia coli as a fusion to the carboxyl terminus of staphylococcal protein A'. The fusion protein was recovered from the bacterial lysate by chromatography on IgG agarose. A 58-kDa protein copurified with the fusion protein in approximately equal amounts. Much less of the 58-kDa protein copurified with a fusion in which the transit peptide was deleted, and it did not copurify with protein A'. The 58 kDa protein was identified as the E. coli groEL gene product with antibodies directed against a homologous mitochondrial heat shock protein. This finding is particularly interesting because a chloroplast protein involved in the assembly of ribulose-1,5-bisphosphate carboxylase/oxygenase also is homologous to the groEL protein. These homologs could modulate protein-protein interactions during folding and assembly of subunits into native complexes. PMID- 2566611 TI - Guanylate cyclase, a cell surface receptor. AB - Guanylate cyclase appears to represent a central member of a diverse family of proteins involved in cell signaling mechanisms including the protein kinases, a low Mr ANP receptor, and possibly adenylate cyclase (based on limited sequence identity with the yeast enzyme). A membrane form of guanylate cyclase represents a new model for cell surface receptors, although such a model was once envisioned for adenylate cyclase (79). In original models for adenylate cyclase, hormone was thought to bind with either the enzyme or with an unknown protein to enhance cyclic AMP production (79). Guanylate cyclase appears to fall into the first adenylate cyclase model where binding of a ligand to an extracellular site on the enzyme transmits a signal to an intracellular catalytic site. The production of cyclic GMP, a second messenger, and of pyrophosphate are then increased. The protein tyrosine kinase family of receptors (80) and possibly another forthcoming family of cell surface receptors containing protein tyrosine phosphatase activity (81-83) contain a single transmembrane domain like guanylate cyclase. Furthermore, the protein tyrosine kinases are activated by ligand binding to the extracellular domain. However, the activation of guanylate cyclase, unlike these cell surface receptors, results in the formation of a low molecular weight second messenger. PMID- 2566612 TI - Characterization and partial purification of a chloride- and calcium-dependent glutamate-binding protein from rat brain. AB - A glutamate-binding protein was solubilized from rat brain synaptic plasma membranes using sodium cholate. Its properties were characterized after addition of exogenous phospholipids and formation of proteoliposomes. Glutamate binding was dependent on calcium and chloride ions with maximal binding at concentrations of 10(-5) M calcium and 10 mM chloride ions. The effects of the two ions were synergistic rather than additive. In addition, glutamate binding was not affected by inhibitors specific for N-methyl-D-aspartate and kainate receptor subtypes, but was inhibited by quisqualate (Ki = 50 microM) and DL-2-amino-4 phosphonobutyrate (Ki = 1.3 mM). Furthermore, binding was abolished by 100 microM 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid and 1 mM dithiothreitol. These properties resemble those of the chloride- and calcium-dependent binding site. Starting from the detergent extract, the glutamate-binding protein was purified 123-fold using fractionated ammonium sulfate precipitation, chromatography on hydroxyapatite and on DEAE-Sephacel as sequential purification steps. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified protein fraction showed two major bands migrating with Mr values of 51,000 and 105,000. The properties of the partially purified binding protein were similar to those of the detergent extract. Glutamate binding to the partially purified protein is not due to a sequestration process or product binding to N-acetylated alpha-linked dipeptidase. Thus, the functional role of the binding protein remains to be established. PMID- 2566613 TI - Somatostatin gene expression in pancreatic islet cells is directed by cell specific DNA control elements and DNA-binding proteins. AB - Somatostatin is a peptide synthesized in the pancreatic islets, nervous system, gastrointestinal tract, and thyroid gland. Factors that control islet cell specific expression of the somatostatin gene were analyzed by expression of fusion genes consisting of 5' rat somatostatin gene sequences linked to coding sequences of the receptor genes, bacterial chloramphenicol acetyltransferase, and human growth hormone. Fusion genes containing 900 and 250 base pairs (bp) of 5' flanking DNA were preferentially expressed at 5-10-fold higher levels in somatostatin-producing islet cell lines, as compared with islet cell lines that produced insulin and glucagon, and in three non-islet cell lines. A deletional mutation consisting of only 65 bp of 5'-flanking sequence of the rat somatostatin gene expressed in all islet cell lines but not in non-islet lines, indicating the existence of a negative-acting islet cell-specific element located between nucleotides -250 and -65. The 65-bp sequence contains the octameric cAMP responsive enhancer (CRE) TGACGTCA (nucleotides -48 to -41). Fine mapping of sequences responsible for islet-specific expression by substitution of synthetic oligonucleotide cassettes revealed full retention of expression by deletion to nucleotides -48 and complete loss of expression at nucleotides -42 of the CRE. Substitution of the 9 bp adjacent 3' to the CRE of the somatostatin gene (nucleotides -40 to -32) with the corresponding sequence located 3' to the CRE of the glucagon gene abolished expression. By gel mobility shift and DNaseI footprinting analyses, proteins in extracts of islet cells bound to the 24 bp including the CRE and downstream adjacent 9 bp (nucleotides -58 to -35). An additional upstream region of DNA was protected from DNase I digestion (nucleotides -110 to -80). Proteins from non-islet cells bound to the region from nucleotides -58 to -35, but patterns of DNase I protection differed from those using proteins from islet cells. These observations indicate that several DNA binding proteins interact with cis-acting elements located between 35 and 58 bp upstream of the transcriptional start site of the rat somatostatin gene to determine islet cell-specific gene expression. CRE-binding protein(s) is ubiquitous among phenotypically different cells, and expression of the somatostatin gene in non-somatostatin-producing islet cells appears to be inhibited by a negative-acting element located upstream of the CRE. PMID- 2566614 TI - Opposite effects of chronic cortisol treatment on pre- and postsynaptic actions of clonidine in pithed rats. AB - 1. Both clinical and experimental studies have shown that chronic elevation of plasma cortisol levels are attended by altered adrenergic receptor function. In the present study we examined the effects of chronic cortisol treatment (25 mg kg 1 day-1 for 7 days by minipumps) on the peripheral cardiovascular responses of pithed, adrenal demedullated vagotomized rats. 2. Chronic cortisol treated rats had higher basal diastolic blood pressures after being pithed, suggesting that sympathetic outflow is not required to sustain elevated peripheral resistance in glucocorticoid-induced hypertension. 3. Whereas alpha 1-adrenoreceptor-mediated diastolic blood pressure responses were unaltered, alpha 2-adrenoreceptor mediated vasopressor responses were potentiated in pithed rats which were chronically treated with cortisol. 4. Elevation of plasma noradrenaline induced by the stimulation of the entire sympathetic outflow of pithed rats was not changed but the clonidine-induced presynaptic alpha 2-adrenoreceptor inhibition of noradrenaline release was attenuated by chronic cortisol treatment. 5. In conclusion, in adrenal demedullated pithed rats the responses mediated by peripheral pre- and postjunctional alpha 2-adrenoreceptors appeared to be affected oppositely after chronic cortisol treatment, suggesting that the pharmacologically homogeneous alpha 2-adrenoreceptor population may be modulated differently by chronic elevation of plasma glucocorticoids. PMID- 2566615 TI - Noradrenaline-induced constriction of large and small coronary arteries in the anaesthetized dog. AB - 1. In the anaesthetized dog external diameter of the left circumflex coronary artery and blood flow through that artery were measured to allow the effect of noradrenaline to be compared in large arteries and resistance vessels. 2. The injection of noradrenaline (0.5 micrograms kg-1) into the coronary artery, after bilateral vagotomy and antagonism of beta-adrenoreceptors, decreased large coronary artery diameter and coronary blood flow. Calculation of resistance in the large coronary artery and in the total left circumflex coronary vascular bed revealed that noradrenaline induced increases of 66% and 89% respectively. 3. Intra-coronary (i.c.) injection of the alpha 1-adrenoreceptor agonist phenylephrine (0.5-2 micrograms kg-1) decreased large coronary artery diameter and coronary blood flow. The alpha 2-adrenoreceptor agonist B-HT 920 (0.5-2 micrograms kg-1 i.c.) decreased coronary blood flow but did not significantly affect large coronary artery diameter. 4. Antagonism of alpha 1-adrenoreceptors with prazosin (10 micrograms kg-1 i.c.) abolished the noradrenaline-induced constriction of the large coronary artery but only partially attenuated the decrease in blood flow. The alpha 2-adrenoreceptor antagonist idazoxan (50 micrograms kg-1 i.c.) partially attenuated the noradrenaline-induced decrease in coronary blood flow but did not affect the large artery constriction. 5. It is concluded that noradrenaline constricts both large and small coronary arteries. Noradrenaline-induced constriction of the resistance vessels is more powerful than in the large artery. The constriction of large arteries in response to noradrenaline is mediated by alpha 1-adrenoreceptors. Postjunctional alpha 1- and alpha 2-adrenoreceptors are both involved in the constriction of the resistance vessels. PMID- 2566616 TI - Stimulation of postjunctional alpha-1 and alpha-2 adrenoreceptors increases proximal urethral perfusion pressure in the pithed rat. AB - 1. In pithed rats, the effects of postjunctional alpha-1 and alpha-2 adrenoreceptor stimulation on prostatomembranous urethral perfusion pressure (UPP) were characterized by using selective adrenoreceptor agonists and antagonists. 2. Dose-dependent increases in UPP were elicited by the intravenous administration of selective alpha-1 and alpha-2 adrenoreceptor agonists, phenylephrine (1.0-30.0 micrograms kg-1) and BHT-933 (10-3000 micrograms kg-1), respectively. 3. The effects of phenylephrine on UPP were antagonized, in a dose related manner, by pretreatment with prazosin (0.1 and 0.5 mg kg-1, iv), a selective alpha-1 adrenoreceptor antagonist. In contrast, pretreatment with rauwolscine 0.1 and 0.5 mg kg-1, iv), a selective alpha-2 adrenoreceptor antagonist, did not alter phenylephrine-induced increases in UPP. 4. BHT-933 elicited increases in UPP that were antagonized, in a dose-related manner, by pretreatment with rauwolscine (0.1 and 0.5 mg kg-1, iv). Neither dose of prazosin altered the effects of BHT-933 on UPP. 5. These results demonstrate increases in the resistance to flow (constriction) through the proximal urethra following selective stimulation of postjunctional alpha-1 and alpha-2 urethral adrenoreceptors. 6. The role of urethral alpha-adrenoreceptor subtypes in the neurohumoral control of the urethra remains to be determined. PMID- 2566618 TI - Simplified procedures for the determination of fenoldopam and its metabolites in human plasma by high-performance liquid chromatography with electrochemical detection: comparison of manual and robotic sample preparation methods. AB - Quantitative analytical methods, based on high-performance liquid chromatography with electrochemical detection, were developed for fenoldopam and its metabolites in human plasma. Two extraction methods, a liquid-liquid extraction method for fenoldopam and its methoxy metabolites and a liquid-solid extraction procedure for the sulfate and glucuronide conjugates of fenoldopam were developed. The extractions can either be performed manually or by robot. The limit of detection for fenoldopam, its sulfate and methoxy metabolites was 0.025, 2 and 0.5 ng/ml, respectively, at a signal to noise ratio of 4. The intra-assay and inter-assay coefficients of variation for both manual and robotic extraction procedures were comparable. These methods were suitably selective and sensitive for pharmacokinetic and metabolic studies of fenoldopam. PMID- 2566617 TI - Rheumatoid arthritis therapy: the slow-acting agents. PMID- 2566619 TI - Comparison of immunofluorescence and enzyme-linked immunosorbent assays for the serology of Hantaan virus infections. AB - Three enzyme-linked immunosorbent assay (ELISA) systems based upon different principles were developed for the serology of Hantaan virus infections and compared with an indirect immunofluorescence assay (IFA). The indirect IFA was carried out with gamma-irradiated Hantaan virus-infected and uninfected Vero E6 cells fixed with ethanol (-70 degrees C) or acetone (20 degrees C) on drop slides and a FITC-coupled sheep anti-human Ig preparation. Atypical staining in the IFA was avoided by using ethanol (-70 degrees C) instead of acetone (20 degrees C) fixation. In the first ELISA ('cell-assay'), Hantaan virus-infected or uninfected Vero E6 cells were used as antigens, which after gamma-irradiation were seeded into microtiter ELISA strips. Serial dilutions of human sera were incubated and specific antibodies were demonstrated with a horseradish peroxidase (HRPO) conjugated sheep anti-human Ig preparation. In the second ELISA ('competition assay') an affinity-purified human Ig preparation was used as a capture antibody for Hantaan virus antigen. After incubation of serial dilutions of human sera with this coat, the reactivity of the affinity purified anti-Hantaan virus Ig coupled to HRPO was determined. In the third ELISA ('complex trapping blocking [CTB]-assay') the same capture antibody was used to react with a mixture of the antigen and serial dilutions of human sera. The reactivity with the same HRPO conjugate was then determined. The results obtained in the respective assay systems with sera from people at risk or suspected of Hantaan virus infection coincided well. The CTB-ELISA proved to be faster and more sensitive than both the other ELISA systems, without giving more non-specific reactions: it detected almost all the IFA positive samples.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566620 TI - Role of thyrotropin receptor antibodies in the development of hyperthyroidism: follow-up studies on nine patients with Graves' disease. AB - TSH binding inhibitor immunoglobulin (TBII) and thyroid-stimulating antibody (TSAb) activities were measured serially for 4-32 months in nine patients before and during development of hyperthyroidism due to Graves' disease. Initially, all were euthyroid, seven had thyroid enlargement, one had proptosis, and seven had high serum titers of antithyroid microsomal antibodies. The occurrence of hyperthyroidism was preceded by detection of both TBII and TSAb in four patients and detection of TSAb alone in four patients. One patient had neither TBII nor TSAb when euthyroid. The mean initial TBII and TSAb activities were 10.2 +/- 15.2% (+/- SD) and 2677 +/- 4620%, respectively, when these patients were euthyroid. When they became hyperthyroid, both TBII and TSAb activities increased in all patients. At that time, TBII was detected in all but one (eight of nine subjects; 88.9%), with a mean activity of 58.8 +/- 23.4% (+/- SD), and TSAb was detected in all nine patients, with a mean value of 4508 +/- 4429%. These findings not only indicate the crucial role of TSH receptor antibodies in the development of hyperthyroidism due to Graves' disease, but also suggest that a certain period of subclinical Graves' disease exists before the onset of overt hyperthyroidism in most patients, in the sense that they have TSH receptor antibodies, especially TSAb, in their serum even though they are euthyroid. PMID- 2566621 TI - Improvement of spermatogenesis after treatment with the antiestrogen tamoxifen in a man with the incomplete androgen insensitivity syndrome. AB - A 32-yr-old man with a history of hypospadias, unilateral cryptorchidism, and pubertal gynecomastia (all surgically corrected) presented with complaints of infertility. Examination revealed scant virilization, recurrence of gynecomastia, small but normal sized testes, small prostate, and oligospermia. His plasma LH, testosterone, dihydrotestosterone, and estradiol levels were high, and his plasma FSH was below the reference range of adult men. An assay of pubic skin fibroblast androgen receptors confirmed the diagnosis of a form of incomplete androgen insensitivity syndrome. Administration of the estrogen receptor antagonist tamoxifen (10 mg, twice daily) induced an increase in plasma FSH greater than that which occurred in six men with idiopathic oligospermia. This man's wife conceived three times during a period of 5 yr, each time after he had received tamoxifen for 12-20 weeks and had considerable improvement of sperm parameters. Conversely, upon cessation of tamoxifen therapy, the semen abnormalities returned. These results indicate that estrogen action impaired this man's fertility, and the impairment could be reversed by administration of an estrogen receptor antagonist. PMID- 2566622 TI - Characterization of Borrelia burgdorferi isolates by restriction endonuclease analysis and DNA hybridization. AB - Genomes of several Borrelia burgdorferi isolates from North America and Europe were characterized by restriction endonuclease analysis and DNA hybridization using labeled B. burgdorferi whole-cell DNA (strain ATCC 35210). Several different restriction and homology patterns were observed among these isolates, indicating genotypic heterogeneity within this genus and species. It was concluded from this study that restriction endonuclease analysis of B. burgdorferi whole-cell DNA may be a reliable and accurate method for identifying strains or genotypes of the Lyme disease agent. PMID- 2566623 TI - Clonal analysis of childhood acute lymphoblastic leukemia with "cytogenetically independent" cell populations. AB - Acute lymphoblastic leukemia (ALL) is generally regarded as a clonal disease in which a single abnormal progenitor cell gives rise to neoplastic progeny. Five of 463 cases of childhood ALL with adequately banded leukemic cells were found to have two cytogenetically independent cell populations. In addition, two of the four cases tested had more than two rearranged immunoglobulin genes and (or) T cell receptor genes. To investigate the clonality of these unusual leukemias, we examined the neoplastic cells for X-linked markers extrinsic to the disease. Leukemic cells from each of the three patients heterozygous for an X-linked, restriction fragment length polymorphism showed a single active parental allele, suggesting that both apparently independent cell populations developed from a common progenitor. These cases provide evidence that leukemogenesis involves a multistep process of mutation and suggest that karyotypic abnormalities may be a late event of malignant transformation. PMID- 2566626 TI - The role of dopaminergic agents and the dopamine receptor in treatment for CHF. PMID- 2566625 TI - Gal-Gal pili vaccines prevent pyelonephritis by piliated Escherichia coli in a murine model. Single-component Gal-Gal pili vaccines prevent pyelonephritis by homologous and heterologous piliated E. coli strains. AB - The initial pathogenic step in nonobstructive Escherichia coli pyelonephritis usually involves the binding of a bacterial adhesin with host uroepithelial glycoprotein receptors containing the D-Gal p alpha 1----4 D-Gal p beta 1 (Gal Gal) moiety. In this study, groups of mice were immunized with Gal-Gal pili and challenged 2 wk later intravesicularly with E. coli strains expressing homologous or heterologous pili. 63 of 129 pili-immunized mice (49%) were protected from subsequent E. coli renal colonization compared with 5 of 85 control mice (6%). Among mice that had E. coli cultured from their right kidney, control mice had greater bacterial colony counts than pili-immunized animals (P less than 0.05). Light microscopic examination of kidneys demonstrated less histopathology among pili immunized mice than among control mice (P less than 0.05). Protection correlated with the presence of specific IgG antibodies in the urine and serum that bind to the major pilin structural polypeptide and not to the Gal-Gal pili tip adhesin per se. These results support the concept that immunization with a bacterial surface-coat constituent can prevent mucosal infection by interfering with colonization. Also Gal-Gal pili of E. coli represent a suitable candidate for immunoprophylaxis against pyelonephritis. PMID- 2566627 TI - A pharmacokinetic rationale for three times daily administration of ethotoin (Peganone). PMID- 2566628 TI - Haloperidol and reduced haloperidol concentrations and psychiatric ratings in schizophrenic patients treated with ascorbic acid. AB - Recent reports have suggested an augmentation by ascorbic acid of haloperidol treatment of schizophrenic patients. This study was designed to examine whether pharmacokinetic interactions between ascorbic acid and haloperidol occur in this population. Eight male inpatients diagnosed as having chronic schizophrenia by DSM-III-R criteria and stabilized on a fixed dose of haloperidol were given oral doses of ascorbic acid, 4.5 grams daily, for 2 weeks in an open trial. Serum concentrations of haloperidol and is metabolite, reduced haloperidol, were measured by high performance liquid chromatography. Psychiatric symptoms were monitored using the Psychiatric Symptom Assessment Scale performed by nursing staff blind to the haloperidol status but not to the ascorbic acid dosage. The addition of ascorbic acid was not associated with any change in psychopathology in this group of patients, nor was there any apparent pharmacokinetic interaction with haloperidol. PMID- 2566629 TI - Buspirone suppression of neuroleptic-induced akathisia: multiple case reports. PMID- 2566624 TI - Cooperative interactions of LFA-1 and Mac-1 with intercellular adhesion molecule 1 in facilitating adherence and transendothelial migration of human neutrophils in vitro. AB - The adherence of human neutrophils to human umbilical vein endothelial cells (HUVEC) is partially dependent on the CD11/CD18 family of glycoproteins on the neutrophil and ICAM-1 on the HUVEC. The CD18 heterodimer involved in this adherence was evaluated in vitro using subunit-specific monoclonal antibodies (MAbs). The adherence of unstimulated neutrophils to IL-1-stimulated HUVEC was significantly inhibited by anti-CD11a but not CD11b MAbs, while the adherence of fMLP-stimulated neutrophils was significantly inhibited by both anti-CD11a and CD11b. Anti-CD11a, but not anti-CD11b MAbs, reduced the adherence of unstimulated neutrophils on purified ICAM-1 to the same low level untreated neutrophils exhibited on a control protein, glycophorin. Stimulation with fMLP significantly increased neutrophil attachment to purified ICAM-1, but not to the control protein. Anti-CD11b MAbs reduced this chemotactically augmented adherence to that of unstimulated neutrophils, and in combination with anti-CD11a MAbs reduced adherence to that on the control protein. The results in this report indicate that unstimulated neutrophils exhibit LFA-1-dependent attachment to ICAM-1, and chemotactic stimulation enhances the attachment of human neutrophils to ICAM-1 by a Mac-1-dependent process. PMID- 2566631 TI - Effects of 1 alpha,25-dihydroxyvitamin D3 on the transglutaminase activity of transformed mouse epidermal cells in culture. AB - Induction of the transglutaminase activity of a transformed mouse epidermal cell line (PAM 212 cells) by 1 alpha,25-dihydroxyvitamin D3 (1 alpha,25-(OH)2-D3), the active form of vitamin D3, was investigated. Addition of 1 alpha,25-(OH)2-D3 to a culture medium stimulated twice the transglutaminase activity at a concentration of 10(-7) M, but vitamin D3, prostaglandin E1, E2, and F2 alpha failed to show this induction. Phorbol 12-o-tetradecanoylphorbol-13-acetate (TPA) and dexamethasone also induced an increase in transglutaminase activity. Exposure to both 1 alpha,25-(OH)2-D3 and retinoic acid caused remarkably synergistic effects on the induction of transglutaminase in the PAM 212 cells. In contrast, simultaneous addition of 1 alpha,25-(OH)2-D3 and TPA was antagonistic and resulted in less than additive induction. Vitamin D3 also showed a similar but lesser effect. These results suggest that 1 alpha,25-(OH)2-D3 induces the transglutaminase activity via mechanisms disparate from those of retinoic acid and modifies epidermal differentiation. PMID- 2566630 TI - Interstitial cells of the adult neocortical white matter are the remnant of the early generated subplate neuron population. AB - The postnatal fate of the first-generated neurons of the cat cerebral cortex was examined. These neurons can be identified uniquely by 3H-thymidine exposure during the week preceding the neurogenesis of cortical layer 6. Previous studies in which 3H-thymidine birthdating at embryonic day 27 (E27) was combined with immunohistochemistry have shown that these neurons are present in large numbers during fetal and early postnatal life within the subplate (future white matter), that they are immunoreactive for the neuron-specific protein MAP2 and for the putative neurotransmitters GABA, NPY, SRIF, and CCK. Here, the same techniques were used to follow the postnatal location and disappearance of the early generated subplate neuron population. At birth (P0), subplate neurons showing immunoreactivity for GABA, NPY, SRIF, or CCK are present in large numbers and at high density within the white matter throughout the neocortex, and the entire population can be observed as a dense MAP2-immunoreactive band situated beneath cortical layer 6. Between P0 and P401 (adulthood), the MAP2-immunostained band disappears so that comparatively few MAP2-immunoreactive neurons remain within the white matter. There is a corresponding decrease in the number and density of neurons stained with antibodies against neurotransmitters. In each instance, these neurons could be double-labeled by the administration of 3H-thymidine at E27, indicating that they are the remnants of the early generated subplate neuron population. The major period of decrease occurs during the first 4 postnatal weeks, and adult values are attained by 5 months. Within the white matter of the lateral gyrus (visual cortex), the density of immunostained neurons decreases dramatically: MAP2, 82%, SRIF, 81%, and NPY, 96%. While SRIF-immunoreactive neurons compose a nearly constant percentage of MAP2-immunoreactive neurons in the white matter between P0 (22%) and P401 (23%), those immunoreactive for NPY decline from 18 to 4%. These changes occur during the same period in which there is less than a twofold increase in white matter area. These observations indicate that the interstitial neurons of the adult neocortical white matter are the oldest neurons of the cerebral cortex since most if not all are derived from the subplate neuron population. In addition, a quantitative analysis suggests that the postnatal decline in subplate neuron density cannot be accounted for solely through dilution by differential growth of the white matter and most likely reflects an absolute decrease in subplate neuron number.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2566633 TI - The use of analgesics for cancer pain. PMID- 2566632 TI - Immunocytochemical localization of L-glutamate decarboxylase and catecholamine synthesizing enzymes in the retroperitoneal sympathetic tissue of the newborn rat. AB - The localization of L-glutamate decarboxylase (GAD), the enzyme synthesizing gamma-aminobutyric acid, was studied in newborn rat retroperitoneal sympathetic tissue, i.e. the main retroperitoneal paraganglion, adrenal medullae and abdominal sympathetic ganglia using the indirect immunofluorescence method. The coexistence of GAD with the catecholamine-synthesizing enzymes tyrosine hydroxylase (TH) and phenylethanolamine N-methyltransferase (PNMT) was analyzed in consecutive sections or by staining one section consecutively with different antisera. GAD immunoreactivity was observed only in some cell types of each organ studied. In the main retroperitoneal paraganglion, the small, intensely TH immunoreactive, paraganglion-type cells were GAD-immunoreactive, while the larger moderately TH-immunoreactive, neuron-like cells were non-reactive for GAD. In the adrenal medulla, GAD immunoreactivity was localized only in the adrenaline synthesizing, PNMT-immunoreactive chromaffin cells. The noradrenaline synthesizing, i.e. the TH-immunoreactive cells with no PNMT immunoreactivity, were non-reactive for GAD. In the abdominal sympathetic ganglia, some small intensely TH-immunoreactive cells were GAD-immunoreactive, while the principal neurons were non-reactive for GAD. These results provide immunohistochemical evidence that GAD is present and is colocalized with catecholamine-synthesizing enzymes in various sympathetic tissues of the newborn rat. The present results indicate that GAD is localized in adrenaline-synthesizing cells of all the sympathetic tissues studied. A fraction of noradrenaline-synthesizing cells of retroperitoneal sympathetic tissues, excluding the adrenal medulla, also contains GAD. PMID- 2566634 TI - Soluble IL-2 receptor in AIDS. Correlation of its serum level with the classification of HIV-induced diseases and its characterization. AB - By using a fluorescence sandwich ELISA, elevated IL-2R levels were detected in the sera from both HIV-infected hemophiliacs and other HIV-infected patients. The serum IL-2R levels were reflective of the classification of HIV-induced diseases by the Centers for Disease Control. Moreover, the IL-2R levels were negatively correlated most prominently with CD4 cell counts, with lymphocyte counts, and with a decrease in the CD4-CD8 ratio but not with either WBC counts or B cell counts. As striking elevations of serum IL-2R were noted in AIDS patients with group IVD infection, the serum IL-2R was purified sequentially by using size exclusion HPLC, high-pressure chromatofocusing, and H48 affinity HPLC. The isoelectric point values of IL-2R were separated into 4.2 and 3.8, whereas the Mr was determined to be only 45 kDa by immunoprecipitation with H48 antibody followed by SDS-PAGE. However, production of cellular and supernatant IL-2R was not elevated in PBMC of patients with AIDS or in any of the 19 HIV-I- or HIV-II infected cell line cells. In contrast, PBMC from patients with adult T cell leukemia and cell line cells that expressed human T cell lymphotropic virus -I or -II produced soluble IL-2R, constitutively. The mechanisms by which serum levels of IL-2R might be elevated in HIV-infected patients are discussed in comparison with that in adult T cell leukemia patients. PMID- 2566635 TI - Linkage analysis of the Bcg gene on mouse chromosome 1. Identification of a tightly linked marker. AB - We have mapped and determined the gene order of five cloned genes in the vicinity of the murine host resistance gene Bcg on mouse chromosome 1. For this, we have used a RFLP-type analysis in panels of 43 recombinant inbred strains, 3 congenic mouse strains, and 186 segregating backcross progeny derived from inbred strains of Bcgr and Bcgs genotypes. The Bcg alleles of segregating animals were established by in vivo infection with Mycobacterium bovis (Bacillus Calmette Guerin) strain Montreal. Genomic DNA prepared from progenitor mouse strains was isolated, digested with restriction endonucleases, and analyzed by Southern blotting to identify strain-specific RFLP for each DNA marker tested. Among a number of DNA markers tested, Len2, Fn, Vil, Alpi, and Achrg were found to co segregate with Bcg in mouse strains congenic for this locus. Detailed segregation analysis of the five markers and Bcg showed that Vil was extremely close to Bcg with no recombinant identified, whereas Fn and Len2 were located 4.5 and 9 cM proximal of Bcg, respectively. Alpi and Achrg mapped 5 and 5.5 cM distal from Bcg, respectively. Pedigree analysis in the recombinant inbred strains and backcross animals indicated the gene order: centromere-Len2-Fn-Vil,Bcg-Alpi Achrg. The tightly linked Vil marker can now be used as an entry point in recombinant genomic DNA libraries to clone sequences overlapping Bcg. This group of five genes flanking Bcg on mouse chromosome 1 is precisely conserved on the telomeric end of the long arm of human chromosome 2q. Our results suggest that a likely location for a putative human homologue to the murine host resistance gene Bcg is the long arm of human chromosome 2 (2q32-qter). PMID- 2566636 TI - Variation in binding of Bacillus sphaericus toxin and wheat germ agglutinin to larval midgut cells of six species of mosquitoes. AB - Bacillus sphaericus toxin labeled with fluorescein isothiocyanate was readily ingested by Culex pipiens, Aedes aegypti, Anopheles stephensi, Anopheles gambiae, Anopheles quadrimaculatus, and Anopheles albimanus larvae. Fluorescent toxin bound to the luminal cell surface in discrete regions of the posterior midgut and gastric caecum in C. pipiens. In Anopheles spp., toxin bound in a variable pattern to these structures and central and anterior midgut as well. The toxin did not bind to midgut cells of A. aegypti. The toxin was internalized in bright fluorescent vesicles in C. pipiens, but was not internalized in Anopheles spp. and appeared to be weakly bound in these larvae, leaking rapidly from the gut surface. The lectin, wheat germ agglutinin, which interferes with binding of the B. sphaericus toxin, bound to the posterior midgut and gastric caecum of all species, but was not internalized. These results suggest that the sugar moiety of the receptor is not solely responsible for specificity of this toxin, and that binding to Culex spp. midgut cells may be highly specific and of high affinity, whereas binding to Anopheles spp. cells may be nonspecific and/or of low affinity. PMID- 2566637 TI - Serum beta 2-microglobulin decreases in patients with AIDS or ARC treated with azidothymidine. AB - Abnormally elevated serum beta 2-microglobulin has been associated with progression of human immunodeficiency virus (HIV) disease and could reflect in vivo HIV activity. We prospectively studied the effect of azidothymidine therapy on serum beta 2-microglobulin concentration in 41 patients with AIDS or AIDS related complex. Median beta 2-microglobulin concentration decreased from 4.02 mg/L before therapy to 3.73 mg/L at week 24 of therapy (P = .016). Individual changes in beta 2-microglobulin during azidothymidine therapy correlated with changes in serum HIV p24 antigen (Spearman, r = .42, P = .007). Also, in a randomized placebo-controlled study, median beta 2-microglobulin concentration decreased after 16 w of therapy in 5 azidothymidine-treated patients compared with levels in 7 placebo-treated controls (P = .05). Serum beta 2-microglobulin appears to be a sensitive marker for in vivo antiretroviral drug activity and may be a better marker than serum p24 antigen for early intervention trials involving antiretroviral agents. PMID- 2566638 TI - Responsiveness of the hamster pancreatic cancer to treatment with microcapsules of D-Trp-6-LH-RH and somatostatin analog RC-160. Histological evidence of improvement. AB - The effect of treatment with D-Trp-6-LH-RH, an agonist of luteinizing hormone releasing hormone (LHRH), and somatostatin analog RC-160 was studied in male Syrian hamsters with N-nitrosobis(2-oxopropyl)amine (BOP)-induced pancreatic carcinoma. The peptides were administered periodically in long-acting microcapsule formulations designed to release controlled doses and maintain continuous blood levels of these analogs. The treatment lasted 60 d. Eighteen wk after administration of BOP, 80% of the animals developed ductal pancreatic adenocarcinomas, typically in multinodular form. Treatment with D-Trp-6-LH-RH resulted in a significant decrease in the tumorous pancreatic weight, and, in 35% of the specimens, changes indicative of histological regression were seen. Similarly, regressive alterations in the tumorous epithelium could be observed in 28% of the tumors in the RC-160 treated group. This regression was not accompanied by accumulation of lymphoid cells and only the epithelial components of the tumors were involved. These data indicate that the analogs D-Trp-6-LH-RH and RC-160 exert antitumoral effects on the experimentally-induced pancreatic cancer. It is unlikely that immunological mechanisms are involved in this response. These inhibitory effects on tumor growth could be mediated by creating a state of sex hormone deprivation of D-Trp-6-LH-RH and by inhibition of the release and/or action of gastrointestinal hormones and growth factors by the somatostatin analog RC-160. PMID- 2566640 TI - The cytoplasmic surface structures of uncoated vesicles in various tissues of rat as revealed by quick-freeze, deep-etching replicas. AB - We examined the cytoplasmic surface structures of the plasmalemmal vesicles in various tissues using quick-freeze, deep-etching replicas, and identified three types of cytoplasmic surface specializations. In addition to the baskets of clathrin-coated vesicles, stripe-surfaced and bumpy-surfaced vesicles could be identified. The striped patterns were clearly observed on the vesicles in endothelial cells and fibroblasts of various tissues, while in epithelial cells such as hepatocytes, renal proximal tubule cells, and superficial cells of the urinary bladder, instead of the striped patterns, rough-surfaced vesicles could be identified. These differences in 'uncoated' vesicles may suggest that there is the tissue specificity of the cytoplasmic surface structures of plasmalemmal vesicles. PMID- 2566639 TI - Clinical usefulness of sialyl SSEA-1 antigen as tumor marker for ovarian cancer as compared with CA125, CA19-9, TPA, IAP, CEA and ferritin. AB - In order to determine the clinical significance of sialyl SSEA-1 antigen, we compared its usefulness as a tumor marker for ovarian cancer with simultaneously measured CA125, CA19-9, TPA, IAP, CEA and ferritin. The sialyl SSEA-1 antigen in serum was measured by radioimmunoassay with an "FH-6" Otsuka Kit. The immunohistochemical localization of sialyl SSEA-1 antigen in ovarian carcinoma tissues was determined by an immunoperoxidase method using FH-6 monoclonal antibody. Among fifty-one patients with ovarian cancer, the incidence of elevated serum levels was 54.9% with sialyl SSEA-1 antigen, 90.2% with CA125, 48.8% with CA19-9, 78.0% with TPA, 73.1% with IAP, 17.1% with CEA and 63.4% with ferritin. On the other hand, among the patients with uterine malignancies and gynecologic benign tumors, the incidence of elevated sialyl SSEA-1 antigen levels in serum was lower than that of other tumour markers. In the patients with ovarian cancer, the serum levels of sialyl SSEA-1 antigen increased in accordance with the advance of the clinical stage and were also correlated with the effect of therapy. In the examination of immunohistochemical localization of sialyl SSEA-1 antigen, a positive reaction occurred in 10 out of 30 ovarian carcinoma specimens. Intense staining appeared in the secretory materials, in the luminal surface of the glands, and in the cytoplasm of cells. Thus, sialyl SSEA-1 antigen appears to be a useful tumor marker for the diagnosis of ovarian cancer, especially when measured simultaneously with CA125, CA19-9, TPA, ferritin and IAP. PMID- 2566641 TI - Aminergic and peptidergic modulation of motor function at an identified neuromuscular junction in Helisoma. AB - Electrophysiological studies suggest that motoneurone B19 in the buccal ganglia of Helisoma makes monosynaptic, cholinergic connections with the supralateral radular tensor (SLT) muscle of the buccal mass. Serotonin (5-HT) and small cardioactive peptide B (SCPB) were found to have peripheral modulatory effects on this motor pathway that are consistent with their previously described central facilitatory effects. Both neurotransmitters, when applied exogenously (10(-6) mol l-1) to isolated buccal ganglion-buccal muscle preparations, potentiated the magnitude of motoneurone B19-evoked muscle contractions (6.3 and 2.7 times, respectively) without affecting excitatory junctional potential (EJP) amplitudes. When applied to single dissociated SLT muscle fibres in cell culture, these modulators had similar effects on acetylcholine (ACh)-evoked muscle fibre shortening, demonstrating that these neuromodulators exert direct actions on the muscle cells. The cardioactive peptide FMRFamide (10(-6) mol l-1), although slightly potentiating muscle contractions in reduced neuromuscular preparations, significantly decreased both ACh-evoked muscle fibre shortening and depolarizing potentials in cultured SLT muscle cells. The differential effects of FMRFamide may, in part, be due to the elimination of interactive effects between multiple neurotransmitters that might exist in semi-intact preparations and in vivo. These results demonstrate that 5-HT, SCPB and FMRFamide in Helisoma can directly modulate the peripheral muscle targets of buccal motoneurones involved in the generation of cyclical feeding behaviour. PMID- 2566642 TI - Clinical biochemistry of sporidesmin natural intoxication (facial eczema) of sheep. AB - The effects of facial eczema, i.e. intoxication by the mycotoxin sporidesmin, were investigated by extensive biochemical screening of serum in 100 controls (A), 31 clinically ill (B) and 219 apparently healthy (C) Manech ewes under field conditions. Dramatic increases of gamma-glutamyltransferase1), alkaline phosphatases, bilirubin, cholesterol, aspartate and alanine aminotransferases, and lactate dehydrogenase confirmed the severity of liver damage in group B, but they were also observed in slightly more than 50% of group C animals. This demonstrated the true extent of the disease, which could be best assessed by the measurement of serum gamma-glutamyltransferase; but since this enzyme was still elevated one year later, it cannot be used as a reliable prognostic parameter. PMID- 2566644 TI - Immunoblot analysis of the serological response in Hantavirus infections. AB - Sera from patients with nephropathia epidemica (NE) or Korean hemorrhagic fever (KHF) were tested for specific antibody response to antigens of Hallnas virus and Hantaan virus strain 76-118. A Vero E6 derived cell line persistently infected with Hallnas virus strain B1, and Vero E6 cells freshly infected with Hantaan virus type strain 76-118 were used as antigens in the immunofluorescence assay (IFA) and the immunoblot. Blots were prepared from whole cell lysates. The convalescent-phase sera of NE patients tested in this study regularly revealed a marked reaction with a 52 kilodalton (Kd) protein of Hallnas virus and a 50 Kd protein of Hantaan virus. A convalescent serum from a patient with Korean hemorrhagic fever and a rat antiserum against Hantaan virus could recognize the 50 Kd band of Hantaan virus but showed no apparent reactivity with the 52 Kd component of Hallnas virus in the standard dilutions. Some sera could additionally identify minor bands in the 55 Kd and/or 67 Kd region of the blots. A one-way cross reactivity between Hantaan and Hallnas viruses was also evident from the results of the immunofluorescence assays in that NE convalescent sera reacted with both viruses, whereas KHF convalescent or anti-Hantaan sera gave strongly positive results with Hantaan virus but only faint reaction with Hallnas virus. PMID- 2566645 TI - Absence of perinatal transmission of blood-borne non-A, non-B hepatitis virus by chimpanzees with acute and chronic infection. AB - Two chimpanzees were born to parents with chronic non-A, non-B hepatitis and remained with their mothers until 12 and 18 months, respectively. The infants were followed from 7 to 8 weeks of age with biweekly or monthly blood samples and with monthly liver biopsies from 4 to 7 months after birth. Another chimpanzee, along with both of its parents, was held throughout the parents' acute infection with non-A, non-B hepatitis; at this time the infant was 14-16 months of age, and it was followed with bi-weekly blood samples and monthly biopsies from the time of potential exposure for 20 months. No abnormalities indicative of non-A, non-B hepatitis were detected in these animals. During the 29 to 35 months of follow up, alanine aminotransferases and gamma glutamyl-transferases (GGPT) levels remained well within normal range for animals held in the same facility. Histologic and electron microscopic examination of liver tissue revealed no abnormalities. PMID- 2566643 TI - Metabolic changes in coho and chinook salmon resulting from acute insufficiency in pancreatic hormones. AB - Acute deficiency in pancreatic peptides (insulin, somatostatin-25, glucagon, and glucagon-like peptide) was invoked for 9-12 hr in coho, Oncorhynchus kisutch, and chinook, O. tshawytscha, salmon by administration of specific antisera raised against purified salmon hormones. Insulin-deficient fish were hyperglycemic, had diminished glycogen content in the liver (Plisetskaya et al., '88a, elevated liver triacylglycerol lipase activity, and higher concentration of plasma triiodothyronine (T3) compared to a control group of fish injected with nonspecific rabbit serum. After immunoneutralization of somatostatin-25, fish remained normoglycemic, with higher liver glycogen content, decreased lipase activity, and elevated plasma levels of insulin, while the levels of T3 declined. The induced deficiency in glucagon family peptides led to comparatively smaller changes: liver glycogen content was increased after anti-glucagon-like peptide (aGLP) injection and transient hyperglycemia was apparent following anti-glucagon (aGLU) administration. Circulating levels of insulin remained unaffected for at least 9 hr following aGLU and aGLP treatments. The velocity of pyruvate kinase at 2.5 mM phosphoenolpyruvate (V2.5) was depressed, especially after the combined administration of aGLU + aGLP. The effectiveness of immunoneutralization experiments was greatly dependent on the particular stage of the fish life cycle. Antisera against fish pancreatic peptides proved to be a suitable tool in the studies of hormonal regulation of fish metabolism. PMID- 2566646 TI - Stimulation of D-2 dopamine receptors decreases the evoked in vitro release of [3H]acetylcholine from rat neostriatum: role of K+ and Ca2+. AB - Reportedly, stimulation of D-2 dopamine receptors inhibits the depolarization induced release of acetylcholine from the neostriatum in a cyclic AMP-independent manner. In the present study, we investigated the role of K+ and Ca2+ in the D-2 receptor-mediated inhibition of evoked [3H]acetylcholine release from rat striatal tissue slices. It is shown that the D-2 receptor-mediated decrease of K+ evoked [3H]acetylcholine release is not influenced by the extracellular Ca2+ concentration. However, increasing extracellular K+, in the presence and absence of Ca2+, markedly attenuates the effect of D-2 stimulation on the K+-evoked [3H]acetylcholine release. Furthermore, it is shown that activation of D-2 receptors in the absence of Ca2+ also inhibits the veratrine-evoked release of [3H]acetylcholine from rat striatum. These results suggest that the D-2 dopamine receptor mediates the decrease of depolarization-induced [3H]acetylcholine release from rat striatum primarily by stimulation of K+ efflux (opening of K+ channels) and inhibition of intracellular Ca2+ mobilization. PMID- 2566647 TI - Glutamate neurotoxicity in culture depends on the presence of glutamine: implications for the role of glial cells in normal and pathological brain development. AB - Glutamate toxicity was studied in neuronal (SC9), glial (WC5), and neuroblastoma glioma hybrid cell lines. In all three cell types, glutamate had a dual effect, depending on the concentration of glutamine in the culture medium. An expected dose-dependent cytotoxicity of the amino acid was observed when cells were cultured in medium containing the standard glutamine concentration (1-4 mM), but when the culture's glutamine content was decreased to 0.15-0.5 mM, glutamate had an apparent opposite, growth-promoting effect. The specificity of glutamate effect was indicated by the following: (a) it was stereospecific, with the L and not the D isomer being active; (b) monosodium aspartate was inactive in the presence of either high or low glutamine; and (c) monosodium glutamate and monopotassium glutamate had a similar dual effect. Furthermore, the glutamate receptor antagonist gamma-glutamylglycine blocked the amino acid cytotoxicity in a dose-dependent fashion. As glial cells are a major source of glutamine in the brain, neuronal-glial co-cultures were used to analyze the possible role of glial cells in glutamate neurotoxicity. It was found that SC9 cells were more sensitive to glutamate when co-cultured with WC5 cells. Continuous depolarization of the SC9 cells with KCl decreased cell number, but glutamate had no additive neurotoxic effect when added with KCl. We suggest that glutamine, glial cells, and neuronal activation play roles in modulating glutamate neurotoxicity, in developing as well as aged brains. It is tempting to speculate also that alterations in the glutamate/glutamine ratio under pathological conditions may take part in the etiology of some neurodegenerative diseases. PMID- 2566648 TI - Release of gamma-[3H]aminobutyric acid from rat olfactory bulb and substantia nigra: differential modulation by glutamic acid. AB - We have studied the glutamate modulation of gamma-[3H]aminobutyric acid ([3H]GABA) release from GABAergic dendrites of the external plexiform layer of the olfactory bulb and from GABAergic axons of the substantia nigra. In the olfactory bulb, [3H]GABA release was induced by high K+ and kainate, and not by aspartate and glutamate alone. However, when the tissue was conditioned by a previous K+ depolarization, glutamate and aspartate caused [3H]GABA release. The effect of glutamate was significantly enhanced when the GABA uptake mechanism was blocked by nipecotic acid. N-Methyl-D-aspartate and quisqualate did not cause [3H]GABA release under the same conditions. The acidic amino acid receptor antagonist 2-amino-4-phosphonobutyric acid and the N-methyl-D-aspartate receptor antagonist 2-amino-5-phosphonovaleric acid significantly inhibited the K+ glutamate- and the kainate-induced [3H]GABA release. Mg2+ (5 mM), which blocks the N-methyl-D-aspartate receptors, significantly inhibited the K+-glutamate induced but not the kainic acid-induced [3H]GABA release. The K+-glutamate stimulated release, but not the K+-stimulated [3H]GABA release, was strongly inhibited by Na+-free solutions or by 300 nM tetrodotoxin. Apparently the glutamate-induced release of [3H]GABA occurs through an interneuron because it is dependent on the presence of nerve conduction. In the substantia nigra no [3H]GABA release was elicited by any of the glutamate agonists tested. The present results clearly differentiate between the effects of glutamate on the release of [3H]GABA from the substantia nigra and from the olfactory bulb.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566649 TI - Frontal cortical and left temporal glutamatergic dysfunction in schizophrenia. AB - Glutamatergic mechanisms have been investigated in postmortem brain samples from schizophrenics and controls. D-[3H]Aspartate binding to glutamate uptake sites was used as a marker for glutamatergic neurones, and [3H]kainate binding for a subclass of postsynaptic glutamate receptors. There were highly significant increases in the binding of both ligands to membranes from orbital frontal cortex on both the left and right sides of schizophrenic brains. The changes are unlikely to be due to antemortem neuroleptic drug treatment, because no similar changes were recorded in other areas. A predicted left-sided reduction in D [3H]aspartate binding was refuted at 5% probability, but not at 10%. Previously reported high concentrations of dopamine in left amygdala were strongly associated with low concentrations of D-[3H]aspartate binding in left polar temporal cortex in the schizophrenics. The findings are compatible with an overabundant glutamatergic innervation of orbital frontal cortex in schizophrenia. The results also suggest that schizophrenia may involve left-sided abnormalities in the relationship between temporal glutamatergic and dopaminergic projections to amygdala. PMID- 2566650 TI - Characterization of [3H]quipazine binding to 5-hydroxytryptamine3 receptors in rat brain membranes. AB - [3H]Quipazine was used to label binding sites in rat brain membranes that display characteristics of a 5-hydroxytryptamine3 (5-HT3) receptor. The radioligand binds with high affinity (KD, 1.2 +/- 0.1 nM) to a saturable population of sites (Bmax, 3.0 +/- 0.4 pmol/g of tissue) that are differentially located in the brain. Specific [3H]quipazine binding is not affected by guanine or adenine nucleotides. ICS 205-930, BRL 43964, Lilly 278584, and zacopride display less than nanomolar affinity for these sites whereas MDL 72222 is approximately one order of magnitude less potent. The pharmacological profile of the binding site is in excellent agreement with that of 5-HT3 receptors characterized in peripheral physiological models. We conclude that [3H]quipazine labels a 5-HT3 receptor in the rat CNS. PMID- 2566651 TI - Role of glial cells for the basal and Ca2+-dependent K+-evoked release of transmitter amino acids investigated by microdialysis. AB - The role of glial cells for the inactivation and synthesis of precursors for amino acid transmitters was studied in the brains of anesthetized rats in vivo using the microdialysis technique. The dialysis probes were inserted stereotactically into each neostriatum. One neostriatum was treated with the gliotoxin fluorocitrate, whereas the contralateral side served as a control. The basal efflux of amino acids, reflecting the extracellular level, was measured as well as the efflux during depolarization with 100 mM K+ in the dialysis stream. The potassium-evoked efflux of transmitter amino acids was calcium dependent and thus considered to reflect release from the transmitter pool. gamma-Aminobutyric acid (GABA) and glutamate release from the treated side was higher than the control value during the first 2-3 h, a result indicating an important role of glial cells in the inactivation of released transmitter. After 6-7 h with fluorocitrate, the release of glutamate was lower than the control value, a result indicating an important role of glial cells in the synthesis of precursors for the releasable pool of glutamate. The role of glutamine for the production of transmitter glutamate and GABA in vivo was further investigated by inhibiting glutamine synthetase with intrastriatally administered methionine sulfoximine. The release of gluatamate into the dialysis probe decreased to 54% of the control value, whereas the release of GABA decreased to 22% of the control value, a result indicating that glutamine may be more important for transmitter GABA than for transmitter glutamate. PMID- 2566652 TI - Presynaptic aspects of cotransmission: relationship between vesicles and neurotransmitters. PMID- 2566653 TI - Observation of cerebral metabolites in an animal model of acute liver failure in vivo: a 1H and 31P nuclear magnetic resonance study. AB - Acute liver failure was induced in rats by a single intragastric dose of carbon tetrachloride. This causes hepatic centrilobular necrosis, as indicated by histological examinations, and produces a large increase in the activity of serum alanine aminotransferase. The plasma NH4+ level (mean +/- SEM) was 123 +/- 10 microM in the control group and 564 +/- 41 microM in animals with acute liver failure (each n = 5). 31P nuclear magnetic resonance (NMR) was used to monitor brain cortical high-energy phosphate compounds, Pi, and intracellular pH. 1H NMR spectroscopy was utilised to detect additional metabolites, including glutamate, glutamine, and lactate. The results show that the forebrain is capable of maintaining normal phosphorus energy metabolite ratios and intracellular pH despite the metabolic challenge by an elevated blood NH4+ level. There was a significant increase in the brain glutamine level and a concomitant decrease in the glutamate level during hyperammonaemia. The brain lactate level increased twofold in rats with acute liver failure. The results indicate that 1H NMR can be used to detect cerebral metabolic changes in this model of hyperammonaemia, and our observations are discussed in relation to compartmentation of NH4+ metabolism. PMID- 2566654 TI - Effects of aging on p- and m-octopamine, catecholamines, and their metabolizing enzymes in the rat. AB - Functions of octopamine in the mammalian brain are still not well known. An important aspect of this problem is the relationship between octopamines and catecholamines. Previous data have shown that their respective ontogenic evolutions are not parallel. Do the changes in brain related to aging also differentially affect these two groups of molecules? In order to check this point, the brain levels of p- and m-octopamine, p-tyramine, noradrenaline, and dopamine, as well as the activities of metabolizing enzymes, were determined in young adult and aging rats (20-26 months). Unlike catecholamines, there is a drastic decrease of p-octopamine after 20 months of age in the hypothalamus and telencephalon. p-Tyramine levels are also lowered. This change appears to be due to a decrease of the aromatic L-amino acid decarboxylase activity. These data, as those of ontogenic studies, confirm that p-octopamine and catecholamine metabolisms may have some independent steps and, moreover, that p-octopamine may have a role in the normal activity of the brain. PMID- 2566655 TI - Direct evidence that excitotoxicity in cultured neurons is mediated via N-methyl D-aspartate (NMDA) as well as non-NMDA receptors. AB - Cultured GABAergic cerebral cortex neurons were exposed to the excitatory amino acid (EAA) L-glutamate, kainate (KA), N-methyl-D-aspartate (NMDA), or RS-alpha amino-3-hydroxy-5-methyl-4-isoxazolopropionate (AMPA). To ensure a constant glutamate concentration in the culture media during the exposure periods, the glutamate uptake inhibitor L-aspartic acid beta-hydroxamate was added at 500 microM to the cultures that were exposed to glutamate. Each of these EAAs was able to induce neurotoxicity. It was not possible to reduce or prevent glutamate induced cytotoxicity by blocking only one of the glutamate receptor subtypes with either the NMDA receptor antagonist D-(-)-2-amino-5-phosphonopentanoate (APV) or with one of the specific non-NMDA antagonists 6-cyano-7-nitroquinoxaline-2,3 dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX). However, if the cultures were exposed simultaneously to glutamate and the antagonists in combination, i.e., APV plus CNQX or APV plus DNQX, the toxicity was completely prevented. Furthermore, CNQX and DNQX were shown to be selective blockers of cytotoxic phenomena induced by non-NMDA glutamate agonists with no effect on NMDA induced cell death. Likewise, APV prevented NMDA-induced cell death without affecting the KA- or AMPA-induced neurotoxicity. It is concluded that EAA dependent neurotoxicity is induced by NMDA as well as non-NMDA receptors. PMID- 2566656 TI - Glutamate release from guinea-pig synaptosomes: stimulation by reuptake-induced depolarization. AB - Glutamate (10-100 microM) reversibly depolarizes guinea-pig cerebral cortical synaptosomes. This does not appear to be because of a conventional autoreceptor. Neither kainate at 1 mM, 100 microM N-methyl-D-aspartate (NMDA), 100 microM L-2 amino-4-phosphonobutanoate (APB), nor 100 microM quisqualate affects the Ca2+ dependent release of glutamate from suboptimally depolarized synaptosomes. However, kainate, quisqualate, and the quisqualate agonists beta-N-oxalylamino-L alanine and alpha-amino-3-hydroxy-5-methylisoxazole propionate cause a slow Ca2+ independent release of glutamate from polarized synaptosomes. However, unlike kainate, quisqualate does not inhibit the acidic amino acid carrier. APB, NMDA, and the NMDA receptor-mediated neurotoxin beta-N-methylamino-L-alanine do not influence Ca2+-independent release at 100 microM. The depolarization of the plasma membrane by glutamate can be mimicked by D-aspartate, can be blocked by the transport inhibitor dihydrokainate, and is accompanied by the net uptake of acidic amino acids. L-Glutamate or D-aspartate at 100 microM increases the cytoplasmic free Ca2+ concentration. D-aspartate at 100 microM causes a Ca2+ dependent release of endogenous glutamate, superimposed on the Ca2+-independent heteroexchange with glutamate through the acidic amino acid carrier. The results suggest that the glutamatergic subpopulation of synaptosomes can be depolarized by exogenous glutamate. PMID- 2566657 TI - Effects of low concentrations of 4-aminopyridine on CA1 pyramidal cells of the hippocampus. AB - 1. Intracellular and extracellular recording techniques were used to study the effects of bath application of 4-aminopyridine (4-AP) on pyramidal cells of the CA1 subfield of rat hippocampal slices maintained in vitro. The concentration of 4-AP used in most experiments was 50 microM. However, similar results were obtained with a concentration ranging from 5 to 100 microM. 2. Following 4-AP application, cells impaled with K-acetate-filled microelectrodes hyperpolarized by an average of 2.6 mV (from -68.7 to -71.3 mV, P less than or equal to 0.01). This change was accompanied by the appearance of high-frequency spontaneous hyperpolarizations. Conversely, when KCl-filled microelectrodes were used, an average depolarization of 5.8 mV [from -73.1 to -67.3 mV, not significant (NS)] associated with the occurrence of repetitive depolarizing potentials was observed. In both cases, these changes were concomitant with a small decrease in membrane input resistance, which was statistically significant only for cells impaled with K-acetate-filled microelectrodes. When synaptic transmission was blocked by tetrodotoxin (TTX), 4-AP induced in cells studied with K-acetate microelectrodes an average depolarization of 2.4 mV (from -62.8 to -60.4 mV, P less than or equal to 0.01) accompanied by a small increase in input resistance (from 32.0 to 35.8 M omega, P less than or equal to 0.05). High-frequency spontaneous potentials failed to occur under these conditions. During 4-AP application, the threshold and the latency of action potentials elicited by a depolarizing current pulse increased in 36% of the neurons studied (n = 14). 3. The amplitude of the stratum (s.) radiatum-induced excitatory postsynaptic potential (EPSP) was augmented by 4-AP. Both the early and late inhibitory postsynaptic potentials (IPSPs) evoked by orthodromic stimuli were also increased in amplitude and duration. In addition, a late (peak latency, 150-600 ms) and long-lasting (duration, 600-1,500 ms) depolarizing potential appeared between the early and the late IPSPs and progressively increased until it partially masked these hyperpolarizations. This long-lasting depolarization (LLD) could also be induced by antidromic stimulation, although in this case it was preceded by an additional, fast-rising, brief depolarization. 4. A similar brief depolarization preceded the orthodromically induced LLD in 69% of the neurons bathed in the presence of 4-AP. The average value of the peak latency of this potential was 62 +/- 27 (SD) ms for orthodromic and 110 +/- 70 ms for antidromic responses.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2566658 TI - A modulatory proctolin-containing neuron (MPN). I. Identification and characterization. AB - The pentapeptide proctolin has been localized previously to the crustacean stomatogastric nervous system and shown to modulate the rhythmic activity of the pyloric network in the stomatogastric ganglion (STG) (Marder et al., 1986; Hooper and Marder, 1987). We have now identified a pair of modulatory proctolin containing neurons (MPNs) that cause proctolin-like modulation of the pyloric rhythm. Individual MPNs were identified by combining intracellular Lucifer yellow dye injection with rhodamine-visualized proctolin immunolabeling. Both MPNs are located in the esophageal nerve and send processes to the STG. Current injection into one MPN influences the second MPN, suggesting that they are electrically coupled. The 2 MPNs have similar effects on the pyloric rhythm of the STG. Intracellular stimulation of a single MPN was sufficient to enhance already active pyloric rhythms and initiated the pyloric rhythm in quiescent preparations. PMID- 2566659 TI - A modulatory proctolin-containing neuron (MPN). II. State-dependent modulation of rhythmic motor activity. AB - The effects of stimulating the modulatory proctolin-containing neurons (MPNs) on the pyloric rhythm of the stomatogastric ganglion of the crab, Cancer borealis, were compared with those produced by exogenously applied proctolin. The effects of both MPN stimulation and proctolin applications depend on the preceding physiological state of the preparation. Both treatments increase the pyloric cycle frequency dramatically in preparations that are slowly cycling, but they have little or no effect on pyloric cycle frequency in preparations that are already rapidly cycling. MPN and proctolin produce maximal pyloric cycle frequencies of about 1.2 Hz, although much faster pyloric frequencies are possible. MPN stimulation and proctolin applications affect the number of impulses fired in each burst by pyloric network neurons. MPN's excitatory actions are longer lasting when a preparation is active than when it is quiescent before stimulation. These data suggest that many of MPN's physiological actions result from its release of proctolin. Small unitary postsynaptic potentials evoked by MPN stimulation in the lateral pyloric neuron may indicate the presence of a second neurotransmitter in MPN. PMID- 2566660 TI - The earliest-generated neurons of the cat cerebral cortex: characterization by MAP2 and neurotransmitter immunohistochemistry during fetal life. AB - The earliest-generated neurons of the cat cerebral cortex have been studied here during development using a combination of 3H-thymidine birthdating with immunohistochemistry for the neuron-specific protein MAP2 or for several neuropeptides/transmitters. These neurons are the first postmitotic cells of the cortex, with birthdates during the 1-week period preceding the genesis of cells of the adult cerebral cortex (Luskin and Shatz, 1985a; Chun et al., 1987). However, they are transient and the majority disappear by adulthood (Luskin and Shatz, 1985a; Chun and Shatz, 1989). When autoradiographic birthdating is combined with MAP2 immunostaining during fetal life, the entire population of these early-generated neurons appears to be stained, resulting in labeled bands above and below the cortical plate. The band above the cortical plate (in the marginal zone) contains early-generated neurons with horizontal morphologies, while the thicker band beneath the cortical plate (within the intermediate zone) contains the somata of early-generated neurons and their elaborate processes that are frequently directed towards the ventricular surface. In view of the correspondence between the location of the early-generated neurons and the MAP2 immunostained band beneath the cortical plate, we suggest that this combined approach can be used to define accurately the subdivision of the intermediate zone known as the subplate. The early-generated neurons are also immunoreactive for GABA, neuropeptide Y (NPY), somatostatin (SRIF), and cholecystokinin (CCK) during fetal life. While GABA, NPY, and SRIF immunostaining could be detected by embryonic day 50 (E50), that for CCK was not found until E60. Moreover, there is a relationship between neuropeptide immunoreactivity and location within the cerebral wall. The marginal-zone neurons are immunoreactive only for CCK. The subplate neurons are immunoreactive for CCK, SRIF, and NPY. Most of those immunoreactive for SRIF tend to be clustered within the upper part of the subplate, while those immunoreactive for NPY tend to be located more deeply. Cells immunoreactive for GABA are more uniformly distributed throughout the cerebral wall. These observations demonstrate directly that the marginal zone and subplate contain peptide- and GABA-immunoreactive neurons that belong to the earliest-generated cell population of the cerebral cortex. The presence of these early-generated neurons, which achieve a remarkable degree of maturity during fetal life, suggests that they perform an essential, yet transient, role in the development of the cerebral cortex. PMID- 2566661 TI - Opiate and peptide inhibition of transmitter release in parasympathetic nerve terminals. AB - Somatostatin, morphine, and opioids inhibit transmitter release at intact neuromuscular junctions between ciliary ganglion neurons and the choroidal smooth muscle of the chick eye. Somatostatin and morphine, however, have no effect on release from terminals on the striated muscle target of the ciliary ganglion, the iris. In neuronal terminals of both the choroid and the iris, a high-affinity Na+ dependent choline uptake-mediated ACh synthesis is present at hatching. Both tissues exhibit a basal release of 3H-ACh which is potentiated severalfold during a 5 minute incubation in 55 mM K+ Tyrodes. Fifty percent of the basal release and 100% of the stimulated release are Ca2+ dependent and probably mediated through N like voltage-dependent Ca2+ channels. Co-incubation of the choroid with 10 microM morphine sulfate blocks approximately 90% of the stimulated release. The same effect is seen with 100 nM somatostatin, 10 microM dynorphin, and 100 microM met enkephalin arginine phenylalanine. Preincubation of the excised choroid with pertussis toxin (200 ng/ml) reverses the inhibitory effects of both morphine and somatostatin. In contrast, 3H-ACh release from terminals in the striated iris is not affected by either morphine or somatostatin at micromolar levels. These results suggest that both opiate and somatostatin receptors are present in the choroid target and that they may act through a final common pathway to modulate ACh release via G proteins. Second messengers such as cyclic AMP or diacylglycerol do not appear to mediate these effects; neither increasing cAMP levels in terminals nor activation of protein kinase C affects evoked release or its inhibition by morphine or other neuromodulators. It is unclear whether endogenous neuromodulation occurs in this system, although somatostatin-like immunoreactivity can be demonstrated in terminals of choroid neurons. PMID- 2566662 TI - Hypobaric-ischemic conditions produce glutamate-like cytopathology in infant rat brain. AB - We present a new animal model of perinatal hypoxic/ischemic brain damage and compare this type of brain damage with the excitotoxic type of damage previously described in the brains of infant rats and monkeys treated systemically with glutamate (Glu). Ten-d-old rats with unilateral occlusion of the common carotid artery were subjected to hypobaric conditions for 75 min and sacrificed 0-4 hr later for light and electron microscopic brain examination. The mortality rate was relatively low (12%), and brain damage was evident ipsilateral to the ligated carotid in 94% of surviving animals 4 hr after termination of the hypobaric event. Regions most frequently affected were the medial habenulum, dentate gyrus, caudate nucleus, frontoparietal neocortices, olfactory tubercle, and several thalamic nuclei. The acute cytopathological changes, primarily edematous degeneration of neuronal dendrites and cell bodies, evolved very rapidly, with some neurons manifesting end-stage necrosis at 0 hr (immediately after hypobaric exposure) and others developing such changes over a 1-4-hr period. We conclude that the neurodegenerative reaction induced in infant rat brain by hypoxia/ischemia is indistinguishable from the excitotoxic type of damage exogenous Glu is known to cause. Moreover, in a companion study (Olney et al., 1989) we show that MK-801, a powerful antagonist of the N-methyl-D-aspartate receptor complex (subtype of Glu receptor), protects against neuronal degeneration in this hypobaric/ischemic model. Our results reinforce other recent evidence suggesting that hypoxic/ischemic brain damage is mediated by endogenous Glu or related excitotoxins. PMID- 2566663 TI - The relationship between the number of synaptic vesicles and the amount of transmitter released. AB - The relationship between the number of synaptic vesicles and the amount of transmitter released from identified synapses was investigated in the dorsal longitudinal flight muscle (DLM) of the temperature-sensitive endocytosis mutant of Drosophila melanogaster, shibirets-1(shi). In the shi fly at 29 degrees C, vesicle recyling is blocked, but transmitter release proceeds normally. Thus, by inducing transmitter release at 29 degrees C, shi synapses gradually become depleted of synaptic vesicles. In this way it was possible to regulate the number of vesicles in a synapse. Intracellular recordings were made from individual fibers of the DLM in shi flies after various periods at 29 degrees C while stimulating at 0.5 Hz. The amplitude of the evoked excitatory junction potential (ejp), gradually decreased with longer exposure and was brought to various levels. The fiber was then rapidly fixed for electron microscopy. The number of vesicles per synapse was compared with the amplitude of the ejp at the time of fixation. It was observed that the smaller the ejp amplitudes became, the fewer vesicles were in the synapses. Also, as the ejp amplitude decreased, an increased number of synapses contained no vesicles. It is concluded that synaptic vesicles are directly involved in the release process. PMID- 2566664 TI - Concomitant characterization of behavioral and striatal neurotransmitter response to amphetamine using in vivo microdialysis. AB - The temporal and dose-related behavioral and striatal monoamine response to amphetamine (AMPH) was examined using in vivo microdialysis in freely moving rats. Extracellular dopamine (DA), serotonin (5-HT), and their metabolites were monitored concomitant with detailed characterization of the locomotor and stereotypy profiles. Consistent with previous results, AMPH (0.5-5.0 mg/kg) induced a rapid dose-dependent increase in DA concentration and decrease in the concentrations of the DA metabolites, DOPAC and HVA. DA and its metabolites exhibited contrasting temporal and dose-related patterns, suggesting that the decline in DA metabolites is functionally dissociated from the AMPH-enhanced DA release. In addition, AMPH at doses of 2.0 mg/kg and greater significantly increased extracellular concentrations of 5-HT, which, in contrast to the changes in dopamine, persisted for only 20-40 min. Comparisons of concentrations of DA and 5-HT for individual animals revealed significant correlations both during baseline and drug response, suggesting a possible functional interdependence between dopaminergic and serotonergic activity in striatum. Dose-response comparisons revealed a significant relationship between AMPH-induced increases in behavioral perseveration and the magnitude and duration of the DA release. However, the temporal patterns of the neurotransmitter response and individual components of stereotypy were not parallel, suggesting that the presence of stereotypies is not associated simply with quantitative differences in striatal DA release. By contrast, some features of the behavioral response were significantly correlated with AMPH-induced changes in striatal 5-HT concentrations. Our results suggest that the behavioral response to AMPH may be influenced by the interaction between levels of DA and 5-HT release, as well as by the state of their respective receptors. PMID- 2566665 TI - Ultrastructural basis for interactions between central opioids and catecholamines. I. Rostral ventrolateral medulla. AB - Opioids and some alpha 2-adrenergic agonists are both known for their potent hypotensive actions following local application to the rostral ventrolateral medulla (RVL), in particular the region containing the C1 adrenergic neurons. We sought to determine whether coexistence and/or synaptic interactions might account for the commonality of cardiovascular responses to opioids and catecholamines in the RVL. Dual light and electron microscopic (EM) immunoperoxidase labeling of a rat monoclonal antibody against the opioid peptide Leucine5 (Leu5)-enkephalin and immunoautoradiographic localization of a rabbit antiserum against the catecholamine synthesizing enzyme tyrosine hydroxylase (TH) were examined in single sections through the RVL of adult colchicine-pretreated rats. Cross-reactivity of the enkephalin antibody was most intense with Leu5 enkephalin. Methionine5-enkephalin as well as dynorphin A, but not beta endorphin, were also recognized by the antisera. By light microscopy, the Leu5 enkephalin-like immunoreactivity (LE-LI) was identified by peroxidase reaction product in perikarya and processes. Most of the perikarya containing LE-LI were located dorsolaterally or ventromedially to those showing immunoautoradiographic labeling for TH. However, a few perikarya appeared to contain both LE-LI and TH immunoreactivity (TH-I) which were difficult to differentiate by light microscopy. By EM, perikarya and dendrites immunoreactive for LE, TH, and both LE and TH were readily distinguishable. Perikarya and dendrites immunoautoradiographically labeled for TH alone were more numerous than those containing LE-LI or TH-I and LE-LI. Axon terminals also were immunolabeled either for one or both reaction products. However, the TH-labeled neurons constituted one of the primary (42% from a total of 118) targets of terminals containing LE LI. Additionally, some of these terminals containing LE-LI shared a common target with TH-labeled terminals. These common target neurons contained either TH-I or TH-I and LE-LI. In most cases, the identified junctions were symmetric and the terminals with LE-LI (0.4-1.2 microns in diameter) contained either (1) a few small clear vesicles (scv's) and numerous intensely immunoreactive large (100-150 nm) dense-core vesicles (dcv's); or (2) many scv's and from 0-6 dcv's of a somewhat smaller (80-120 nm) diameter. The latter type of terminal was more consistently dually labeled for TH. The remaining terminals containing LE-LI formed synaptic junctions with unlabeled perikarya or dendrites (32%), were in apposition to other unlabeled as well as TH or LE- and TH-containing terminals (4%) or were without recognizable specializations within the plane of section (22%).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2566666 TI - Reactions between dipeptidyl peptidase IV and diacyl hydroxylamines: mechanistic investigations. AB - Kinetics of inactivation of dipeptidyl peptidase IV (DP IV, EC 3.4.14.5) by N peptidyl-O-(4-nitrobenzoyl) hydroxylamines and their enzyme-catalyzed hydrolysis were followed using independent monitoring methods, all giving similar efficiency ratios of Kcat/Kinact. Different temperature dependences of the DP IV inactivation and enzyme-catalyzed hydrolysis provide evidence of independent rate determining steps for both reactions. Activation parameters of inactivation are similar to those of spontaneous decomposition of the compounds, suggesting a mechanistic relationship. Investigation of DP IV-inactivation, DP IV-catalyzed hydrolysis of N-Ala-Pro-O-Bz(4-NO2) and the decomposition of the suicide substrate in H2O and D2O gave solvent isotope effects of 4.65, 2.54 and 1.02, respectively. A proton inventory of the inactivation reaction indicates involvement of more than one proton in the formation or breakdown of its transition state. The linear proton inventory found for the hydrolytic reaction is consistent with one proton transition in the rate determining step and resembles the rate limiting deacylation of Ala-Pro-DP IV. The hypothetical reaction model now locates splitting in both reactions prior to formation of a covalent intermediate during the catalytic cycle. PMID- 2566667 TI - Administration of intraspinal analgesia in the home care setting. AB - Intraspinal analgesia can provide relief to patients suffering from intractable pain. Nurses following patients receiving intraspinal analgesia need to be familiar with care of the long-term intraspinal catheter, side effects that occur with spinal analgesia, interventions with acute tolerance, and use of drug combinations in the intraspinal infusion. Eighteen patients were observed for 1 year, using the epidural portal system. This method of intraspinal delivery permitted long-term epidural access without complications of system breakdown or catheter migration. With intraspinal infusions, the patient remained alert and could die at home with dignity, free from pain. PMID- 2566668 TI - Progress in the identification of the heterozygote in phenylketonuria. PMID- 2566669 TI - Sulfasalazine-induced hepatotoxicity in children with inflammatory bowel disease. AB - A case of severe sulfasalazine-induced hepatotoxicity is reported in a 14-year old girl with Crohn's disease. Fourteen days after beginning sulfasalazine, she developed a systemic reaction characterized by high fever, maculopapular rash, lymphadenopathy, abdominal pain, and malaise, with tender hepatomegaly and elevated liver functions, leukemoid reaction, with eosinophilia, and immune complexes. She responded promptly to high-dose intravenous steroids with complete recovery. This case was compared to the three similar pediatric cases in the literature. The latent period was 11-19 days between sulfasalazine therapy and the onset of high fever, generalized lymphadenopathy, and pruritic maculopapular rash that spared palms and soles and resolved with desquamation. All children had hepatotoxicity indicated by tender hepatomegaly, elevated liver enzymes, and histologic inflammation on liver biopsy. The similarity between these features and those of other sulfa-induced hepatotoxic reactions suggests that the sulfapyridine moiety is the etiologic agent in these hypersensitivity reactions. Those children with circulating immune complexes responded well to steroids. Proper therapy for this rare but severe idiosyncratic reaction includes prompt recognition and discontinuation of sulfasalazine, and high-dose corticosteroid therapy. PMID- 2566670 TI - Genetic counseling in X-linked retinitis pigmentosa. AB - The current status regarding genetic counseling in X-linked retinitis pigmentosa (XLRP) is reviewed. XLRP is the most severe form of retinitis pigmentosa (RP) and leads to blindness in the third or fourth decade of life. The biochemical basis of the disease is not known. Until now genetic counseling in this disease has been dependent on simple Mendelian laws of inheritance and the detection of carriers by clinical and electrodiagnostic means. The limitations with regard to carrier detection are discussed. With the recent advances made in recombinant DNA technology, genetic counseling has come to play an important part in the management of XLRP. The methods of DNA technology and their application to localizing the XLRP gene on the X chromosome are reviewed. The discovery of DNA linkage markers known as restriction fragment length polymorphisms (RFLPs) allow a marker closely linked to a disease gene to be followed through succeeding generations in an affected family. Since linkage studies suggest two XLRP loci, carrier detection and prenatal diagnosis of the disease still remain problematic. PMID- 2566671 TI - Transverse testicular ectopia. AB - In transverse testicular ectopia, both testes are present in the same hemiscrotum. We report such a case and review the possible mechanism of this anomaly, the differential diagnosis, recognition, and management. PMID- 2566672 TI - Analysis of the serum antibody responses to type 1 and type 2 fimbriae in mice immunized with Actinomyces viscosus T14V. AB - The antibody responses of inbred mice immunized with Actinomyces viscosus T14V cells were analyzed using enzyme-linked immunoabsorbent assays (ELISAs) for measuring serum antibodies reactive with A. viscosus T14V cells and type 1 and type 2 fimbriae. In A/J mice immunized intraperitoneally on days 0 and 14, the serum antibody responses approached peak levels during d 19-35, and a dose of 10(8) cells/injection elicited optimal responses. Analysis of the responses of three genetically diverse strains of inbred mice indicated striking variations in the amounts of anti-type 1 (6.5-fold) and anti-type 2 (14.3-fold) antibodies elicited. The observed variations in the magnitude of the anti-fimbrial antibody responses are theoretically of sufficient magnitude to account for significant differences between mouse strains in their ability to inhibit adherence of A. viscosus T14V to saliva-coated hydroxyapatite and other bacteria. These studies provide a model with which the effects of variations in anti-fimbrial antibody responses on bacterial adherence may be analyzed. PMID- 2566674 TI - A simple and rapid liquid chromatographic method for the determination of major metabolites of sulfasalazine in biological fluids. AB - A simple and rapid assay for quantitation of sulfasalazine metabolites in rat urine and plasma was developed using high-performance liquid chromatography (HPLC). The method involves dilution of urine or plasma samples (0.1 mL) with methanol for protein precipitation, followed by mixing and centrifugation at 10,000 x g. Chromatography was accomplished with a reversed-phase ODS C-18 column (5 mu; 4.6 x 250 mm). The mobile phase consisted of 20% methanol in 5.0 mM phosphate buffer (pH 6.0), with 0.5 mM tetrabutylammonium chloride as an ion pairing agent. The flow rate was 1.7 mL/min. An injection volume of 30 microL was used and the metabolites were quantitated by an ultraviolet detector at 254 nm. Benzamide was used as the internal standard. This method is linear in the range of 0.5 to 25 micrograms/mL for 5-aminosalicylic acid (5-ASA), acetylsulfapyridine (Ac-SP), and acetyl-5-aminosalicylic acid (Ac-5-ASA), and from 0.25 to 25 micrograms/mL for sulfapyridine (SP). The percent relative standard deviation ranged from 1 to 7.9% for the metabolite standard curves and precision studies. The limit of detection for 5-ASA, Ac-SP, and Ac-5-ASA is 100 ng/mL, and for SP is 50 ng/mL, in both urine and plasma. This method is rapid, precise, and accurate, and has been used to determine sulfasalazine metabolites in individual rat plasma and urine samples following an oral dose of 60 mg/kg of sulfasalazine. PMID- 2566673 TI - Neopterin: a predictive marker of acquired immune deficiency syndrome in human immunodeficiency virus infection. AB - In 79 homosexual men infected with human immunodeficiency virus (HIV), urinary neopterin was significantly higher as compared with 70 HIV-seronegative men in the same cohort (p less than 0.0001). This highly significant association was found both for naturally occurring oxidized (native) neopterin and for total neopterin (native plus chemically oxidized forms). In prospective follow-up for 18 months, the odds that the acquired immune deficiency syndrome (AIDS) would develop were elevated 25-fold among those whose native neopterin levels were in the highest quartile compared with those with lower neopterin levels, and the corresponding odds ratio for total neopterin was 7.8. Logistic regression analyses indicated that neopterin added useful information to T4-cell count in predicting AIDS onset and that both are statistically significant in the multivariate model. A cross-sectional survey revealed that neopterin levels were correlated with number of receptive anal intercourse partners in the year before HIV seroconversion (r = 0.60, p = 0.0005). Since neopterin may serve as a marker of monocyte/macrophage activation by soluble factors such as gamma-interferon, these data support a growing body of virologic and immunologic evidence that highlights the important role of the monocyte/macrophage in the pathogenesis of AIDS. PMID- 2566675 TI - Phosphodiesterase isozyme inhibition and the potentiation by zaprinast of endothelium-derived relaxing factor and guanylate cyclase stimulating agents in vascular smooth muscle. AB - We have examined the interaction of zaprinast with mediators of guanylate cyclase on the relaxation of aortic smooth muscle. Zaprinast, a selective inhibitor of the low Km-cyclic GMP (cGMP) phosphodiesterase [low Km cGMP phosphodiesterase (PDE)], was equally effective in relaxing phenylephrine-contracted aortas from spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) with an intact endothelium [EC50 = 7.6 (3.5-16.6) microM vs. 9.3 (4.1-21.3) microM, respectively]. In contrast, the vasorelaxant activity of zaprinast in intact and denuded phenylephrine-contracted guinea pig aortas, as well as denuded (SHR and WKY) aortas was minimal. Sodium nitroprusside and atriopeptin II were significantly (P less than .05) more potent as vasorelaxants in denuded SHR aortas when compared with denuded aortas from WKY. Pretreatment with zaprinast potentiated the vasorelaxant potency of sodium nitroprusside in both SHR and WKY aortas whereas atriopeptin II responses were potentiated only in WKY aortas. In studies with the low Km cGMP PDE, isolated via DEAE column chromatography, the apparent Km for cGMP and potency of zaprinast were approximately 2-fold greater (P less than .05) in WKY when compared with the same PDE isozyme isolated from SHR aortic preparations. However, the Vmax (picomoles per milligram per minute) for cGMP hydrolysis was greater in SHR than in WKY. In conclusion, these data show that, although there are no apparent differences in the influence of spontaneously released endothelium-derived relaxing factor from SHR and WKY aortas, reactivity differences to other agents known to stimulate guanylate cyclase activity exist between SHR and WKY denuded aortas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566676 TI - Differential modulation by [D-Pen2, D-Pen5]enkephalin and dynorphin A-(1-17) of the inhibitory bladder motility effects of selected mu agonists in vivo. AB - The possibility that the delta agonist, [D-Pen2, D-Pen5]enkephalin (DPDPE) and the putative endogenous kappa agonist, dynorphin A-(1-17) could differentially modulate the effects of a group of chemically diverse mu agonists was evaluated using inhibition of volume-induced contractions of the rat urinary bladder as a model of central nervous system opioid receptor function in vivo. Intracerebroventricular administration of equieffective doses of the mu agonists [D-Ala2, NMPhe4, Gly-ol]enkephalin (DAMGO), [N-MePhe3, D-Pro4]enkephalin (PL017), morphine, normorphine, sufentanil, etorphine, phenazocine, meperidine and methadone inhibited spontaneous bladder contractions for approximately 20 to 30 min. Low doses of DPDPE or dynorphin A-(1-17) failed to affect spontaneous bladder contractions; higher doses of DPDPE (greater than 15.5 nmol) and dynorphin A-(1-17) (i.e., greater than 3.7 nmol), inhibited bladder contractions. When coadministered i.c.v., DPDPE displaced the morphine dose-response line to the left and also potentiated the effects of normorphine and etorphine. In contrast, DPDPE failed to alter the actions of equieffective doses of DAGO, PL017, meperidine, methadone, phenazocine or sufentanil. The potentiation of the effects of morphine by DPDPE were prevented by i.c.v. coadministration of the delta antagonist, ICI 174,864 (N,N-diallyl-Tyr-Aib-Aib-Phe-Leu-OH); at the dose tested, the delta antagonist had no agonist effects alone and did not antagonize the effects of morphine directly. Furthermore, the agonist effects of morphine were potentiated by several different doses of DPDPE. Administration of i.c.v. dynorphin A-(1-17) produced a rightward displacement of the morphine dose response line and also antagonized the effects of normorphine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566677 TI - Effect of continuous exposure to selective D1 and D2 dopaminergic agonists on rotational behavior in supersensitive mice. AB - The effects of continuous exposure to selective dopaminergic agonists were examined in mice with unilateral 6-hydroxydopamine-induced lesions of the corpus striatum. Continuously infusing the D1 agonists, SKF 38393, SKF 75670 and Cy 208 243 with the use of implanted Alzet minipumps initially produced rotational behavior, but this effect decreased during the first 2 days and then stopped completely during days 3 to 7 of drug infusion. Infusion of the D2 agonists quinpirole and N-0437 also produced rotational behavior but, in contrast to the results seen with the D1 agonists, the rotational response remained present throughout the 7 days of drug exposure. The desensitization produced by continuous exposure to SKF 38393 was selective for the D1 system, as animals exposed continuously to SKF 38393 failed to rotate to an acute challenge dose of SKF 38393 but had a normal rotational response to quinpirole. SKF 75670 and CY 208-243 were less selective than SKF 38393; continuous exposure to SKF 75670 and CY 208-243 decreased the response to an acute injection of D1-agonists by 98 and 95%, respectively, and to that of D2-agonists by 64 and 38%, respectively. Infusing the peripherally acting D1 agonist, fenoldopam, or the inactive isomer ( )-SKF 38393 failed to produce desensitization, suggesting that SKF 38393-induced desensitization is produced by an action at D1 receptors within the central nervous system. These results demonstrate that the D1 system can be desensitized independently from the D2 system and that there are different mechanisms for the long term regulation of D1 and D2 dopaminergic systems. PMID- 2566678 TI - Alpha adrenoceptor agonist stimulation of oxygen consumption in rat proximal and distal nephrons. AB - Selective alpha-1 and alpha-2 adrenergic agonists were used to test the hypothesis that both receptor subtypes increase transcellular Na+ transport in the nephron. Oxygen consumption (QO2) was used as an index of transcellular transport and provided a continuous dynamic record of the tubules' response to an agonist. Both alpha-1 and alpha-2 adrenoceptor agonists produced a linear dose related increase in QO2 at a steeper slope than the control in proximal and distal nephron segments. Stimulation of QO2 by the adrenergic agonists did not occur in the presence of ouabain and did not exceed the maximal respiratory rate achieved with nystatin. The response was determined to be a receptor-mediated increase in the ouabain-sensitive component of respiration. An inactive stereoisomer of epinephrine produced no effect, and adrenergic antagonists inhibited the stimulation by the respective agonists. Adrenergic agonists stimulated QO2 in proximal segments to a much greater degree than observed with suspensions of distal segments. These results are consistent with the density of adrenoceptors on nephrons reported in radioligand binding and autoradiographic studies. The alpha-1 agonists, cirazoline and phenylephrine, had similar dose response curves and stimulated proximal tubules more than distal tubules. The alpha-2 agonists, guanabenz, UK 14,304 and B-HT 933, equivalently stimulated QO2 in distal tubules, but a spectrum of enhanced oxygen consumption was observed in proximal nephrons. Although both alpha adrenoceptor subtypes increased QO2 in proximal and distal nephrons, the mechanisms are likely to be different. PMID- 2566679 TI - Novel peptidic mu opioid antagonists: pharmacologic characterization in vitro and in vivo. AB - A series of six synthetic octapeptides, structurally related to somatostatin, demonstrate high affinity and selectivity for mu opioid receptors in radioligand binding assays. The compounds, D-Phe-Cys-Tyr-D-Trp-Lys-Thr-Pen-Thr-NH2 (CTP), D Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2 (CTOP), D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr NH2 (CTAP), D-tetrahydroisoquinoline carboxylic acid (D-Tic)-Cys-Tyr-D-Trp-Lys Thr-Pen-Thr-NH2 (D-Tic-CTP), D-Tic-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2 (D-Tic-CTOP) and D-Tic-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2 (D-Tic-CTAP), were tested in vitro and in vivo for agonist and antagonist potency and selectivity. In vitro bioassays included the guinea pig ileum, mouse vas deferens and rabbit vas deferens. In vivo tests included hotplate antinociception and gastrointestinal transit inhibition, performed in mice. In vitro, all six derivatives were competitive, highly selective mu antagonists (pA2 values from 6.4-7.9). The compounds demonstrated varying degrees of intrinsic agonist activity especially in the mouse vas deferens, the least active being CTAP and D-Tic-CTAP, which showed no mu or kappa agonist actions, and delta activity only at very high (greater than 3 microM) concentrations. In vivo, none of these compounds showed antinociceptive actions when administered i.c.v. in mice. All were competitive mu antagonists in the hotplate antinociception test.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566680 TI - Discriminative stimulus effects of mu and kappa opioids in the pigeon: analysis of the effects of full and partial mu and kappa agonists. AB - Pigeons were trained to discriminate a dose of either 0.01 mg/kg of bremazocine or 0.05 mg/kg of fentanyl from water using a two-key drug discrimination procedure. During tests of substitution, the selective kappa-opioid agonists bremazocine, U50, 488 and tifluadom substituted for the bremazocine stimulus, whereas the less selective kappa-opioid agonists ethylketocyclazocine, levallorphan, proxorphan and nalorphine substituted for the fentanyl stimulus. The full mu-opioid agonists fentanyl, morphine, I-methadone and levorphanol, as well as the partial agonists nalbuphine, butorphanol and buprenorphine, substituted for the fentanyl stimulus. Compounds with partial-opioid agonist effects, namely nalbuphine, butorphanol, buprenorphine, proxorphan, levallorphan and nalorphine, produced 50% fentanyl-appropriate responding at doses 25 to 369.2 times smaller than the doses required to decrease response rates to 50% of control values. In contrast, the full mu-opioid agonists fentanyl, morphine, I methadone and levorphanol produced 50% fentanyl-appropriate responding at doses only 1.3 to 10.9 times smaller than those required to decrease response rates by 50%. During tests of antagonism, both naloxone and Mr2266 produced a dose dependent attenuation of the stimulus effects of bremazocine and fentanyl, whereas beta-funaltrexamine antagonized the stimulus effects of fentanyl but not bremazocine. Although bremazocine has been reported to have mu-opioid antagonist effects, it failed to antagonize the stimulus effects of the training dose of fentanyl. The present investigation establishes further that pigeons can discriminate selective kappa-opioid agonists from mu-opioid agonists and that in pigeons the classification of numerous opioid compounds on the basis of their kappa-like or mu-like stimulus effects differ from those in rat and monkey. In addition, under the drug discrimination procedure the actions of compounds classified as partial-opioid agonists can be differentiated from those of full mu opioid agonists on the basis of the ratio of the dose required to engender fentanyl-like stimulus effects to the dose required to reduce response rates.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566681 TI - Human fat cell alpha-2 adrenoceptors. I. Functional exploration and pharmacological definition with selected alpha-2 agonists and antagonists. AB - This study was undertaken to investigate more fully the pharmacological characteristics of the human fat cell alpha-2 adrenoceptor. Biological assays were performed on intact isolated fat cells while radioligand binding studies were carried out with [3H]yohimbine in membranes. These pharmacological studies brought: 1) a critical definition of the limits of the experimental conditions required for the exploration of alpha-2 adrenergic responsiveness on human fat cells and membranes; 2) an improvement in the pharmacological definition of the human fat cell postsynaptic alpha-2 adrenoceptor. Among alpha-2 agonists, UK 14,304 was the most potent and the relative order of potency was: UK-14,304 greater than p-aminoclonidine greater than clonidine = B-HT 920 greater than rilmenidine. For alpha-2 antagonists, the potency order was: yohimbine greater than idazoxan greater than SK&F-86,466 much greater than benextramine; 3) a description of the impact of benextramine (irreversible alpha-1/alpha-2 antagonist) on human fat cell alpha-2 adrenergic receptors and on human fat cell function; the drug inactivates the alpha-2 adrenergic receptors with a minor impact on beta adrenergic receptors and without noticeable alterations of fat cell function as assessed by preservation of beta adrenergic and Al-adenosine receptor-mediated lipolytic responses; and 4) a definition of the relationship existing between alpha-2 adrenergic receptor occupancy, inhibition of adenylate cyclase activity and antilipolysis with full and partial agonists. The existence of a receptor reserve must be taken into account when evaluating alpha-2 adrenergic receptor distribution and regulation of human fat cells. PMID- 2566682 TI - Human fat cell alpha-2 adrenoceptors. II. Comparative study of partial and full agonist binding parameters using [3H]clonidine and [3H]UK-14,304. AB - Binding studies were carried out on human fat cell membranes with the major radioligands available for alpha-2 adrenergic receptor identification: the antagonist [3H]yohimbine, the partial agonist [3H]clonidine ([3H]CLO) and the full agonist radioligand [3H]UK-14,304 ([3H]UK). Binding approaches performed with [3H]UK and [3H]CLO; two imidazoline derivatives exhibiting full and partial agonist properties, respectively, in biological assays clearly indicate that: 1) partial and full agonists label an equivalent number of binding sites corresponding to the high affinity form of the alpha-2 receptor; 2) there is some correlation between the KiH/KiL ratio defined in competition of [3H]yohimbine binding and the intrinsic activity defined in biological assays; 3) differences exist between the dissociation of the full-agonist ([3H]UK) and the partial agonist ([3H]CLO); 4) the interaction of the full agonist with the alpha-2 receptor promotes the formation of an agonist-alpha-2 receptor-Gi protein complex (HRGi) complex which is more stable than that obtained with the partial agonist as objectivated by the sensitivity to the effects of guanosine 5' (imido)triphosphate and N-ethylmaleimide; 5) the full-agonist is characterized by a "tight agonist binding" which is not observed with the partial agonist. From a functional point of view, the lower biological activity of the partial alpha-2 agonist could be explained by the formation of more labile HRGi complexes having weaker stability by comparison with the full agonist agents which promote stronger HRGi complexes and sustained activity. PMID- 2566683 TI - Metabolic changes in glutathione and metallothionein in newborn rat liver. AB - Metallothionein (MT) and glutathione (GSH) both contain 30% cysteine and they have distinct developmental profiles in perinatal rat liver. The metabolic relationships between these two cysteine pools were investigated in newborn rats under various experimental conditions. Injection of 2-day-old rat pups with buthionine sulfoximine, phorone or diethylmaleate decreased hepatic GSH levels without any change in high basal levels of MT or zinc. Similarly injection of L oxothiazolidine carboxylate increased hepatic GSH levels but no changes in MT or zinc levels were observed. Administration of buthionine sulfoximine in drinking water to pregnant rats from day 14 of gestation decreased hepatic GSH concentrations in both the dams and pups with little change observed in neonatal hepatic zinc and MT levels or in gamma-glutamyltranspeptidase activity. The induction of MT synthesis by zinc salts in newborn rats was not affected by the in utero reduction of GSH levels. Although maternal hepatic GSH levels can be decreased by a sulfhydryl-deficient diet, no changes were observed in GSH, MT or zinc levels in newborn rat liver. Reduction of perinatal hepatic MT levels by in utero zinc deficiency had little effect on GSH levels. However, inhibition of the cystathionase pathway in newborn rats with propargylglycine decreased hepatic levels of MT, zinc and GSH. The results suggest that whereas there is little interaction between these two pools of cysteine, inhibition of cystathionase activity can decrease hepatic concentrations of both GSH and MT. PMID- 2566684 TI - Potentiation of 3-methylcholanthrene induction of rat hepatic cytochrome P450IA1 by dexamethasone in vivo. AB - The potentiation of the expression of polycyclic aromatic hydrocarbon-inducible cytochromes P450IA1 and P450IA2, and phenobarbital-inducible cytochrome P450IIB1 and NADPH-cytochrome P-450 reductase by dexamethasone in vivo was investigated using adrenalectomized and sham-operated rats. Hepatic cytochrome P450IA1 activity (determined by ethoxyresorufin O-deethylase activity) was induced maximally (30-fold) in both sham-operated and adrenalectomized rats 24 hr after a single injection of 3-methylcholanthrene (10 mg/kg). Dexamethasone (10 mg/kg) increased hepatic tyrosine aminotransferase activity 3- to 4-fold, but had little or no effect on ethoxyresorufin O-deethylase activity. However, dexamethasone potentiated the 3-methylcholanthrene-dependent induction of hepatic P450IA1 activity 2- to 3-fold in adrenalectomized rats and 1.5- to 2-fold in sham operated rats when administered concomitantly with 3-methylcholanthrene (P less than .05). The dose of dexamethasone required to potentiate 3-methylcholanthrene induction of P450IA1 activity (greater than 1 mg/kg) correlated well with the dose required to induce hepatic tyrosine aminotransferase activity; a marker of glucocorticoid action. Potentiation of 3-methylcholanthrene induction of P450IA1 activity in the rat appears to be tissue specific in that dexamethasone was found not to potentiate 3-methylcholanthrene induction of P450IA1 activity of rat lung or kidney. Cytochrome P450IA1 content induced by 3-methylcholanthrene in the liver was also potentiated 2- to 3-fold by dexamethasone. In contrast, potentiation of 3-methylcholanthrene induction of cytochrome P450IA2 content was not observed. Western blot analyses indicate that the increase in monooxygenase activity induced by 3-methylcholanthrene or 3-methylcholanthrene plus dexamethasone corresponds well only with the induction of cytochrome P450IA1 protein content.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566685 TI - Inhibition of aminopeptidases by peptides containing ketomethylene and hydroxyethylene amide bond replacements. AB - Inhibitors of aminopeptidase enzymes have been prepared by the synthesis of peptide substrate analogues in which the scissile amide bond has been replaced with the hydrolytically stable ketomethylene (-COCH2-) and hydroxyethylene [ CH(OH)CH2-] functionalities. Two synthetic strategies were used to prepare the inhibitors, and the advantages and disadvantages of each are discussed. The synthesis of peptides that contain the hydroxyethylene isostere was complicated by competing lactone and lactam formation, and attempts to prepare free N terminal dipeptide hydroxyethylene isostere derivatives were unsuccessful. All ketomethylene isosteres examined were weak inhibitors of both leucine aminopeptidase and aminopeptidase M. However, the ketomethylene inhibitor LysK(RS)Phe (58) (Ki = 4 nM) is a potent inhibitor comparable to the natural product, arphamenine A (ArgKPhe; Ki = 2.5 nM). Normal Michaelis-Menten kinetics for inhibition of membrane leucine aminopeptidase are observed in the absence of magnesium ion, but nonlinear kinetics were obtained in the presence of Mg2+. PMID- 2566686 TI - Mark-recapture studies on the Culiseta (Diptera: Culicidae) vectors of eastern equine encephalitis virus. AB - Natural populations of Culiseta melanura (Coquillett) and Cs. morsitans (Theobald) were marked and released at three sites associated with the Toad Harbor-Big Bay Swamp eastern equine encephalitis (EEE) virus focus in central New York. Collections of Culiseta from surveillance sites within 12.8 km of the release sites were screened for marked specimens. A total of 172 marked specimens were recaptured. The majority of recaptured mosquitoes were collected at sites associated with the swamp complex, but some were collected at seven of the eight recapture sites, up to 9.8 km from a release site. The distances traveled by 103 recaptured individuals were used to calculate the mean distance traveled and flight ranges for sexes and cohorts of both species. The mean distance traveled for females that were marked and released from resting shelters was 4 km for Cs. melanura and 5 km for Cs. morsitans. The mean distance traveled for females marked and released from a CDC + CO2 light trap was 9 km and 8 km for Cs. melanura and Cs. morsitans, respectively. The flight ranges of these species overlap the EEE epizootic zone, and the results of these studies support the hypothesis that these species are involved in the transfer of EEE virus from swamp to upland habitats. PMID- 2566687 TI - Culicidae (Diptera) mortality resulting from insecticide aerosols compared with mortality from droplets on sentinel cages. AB - The mortality of female Aedes aegypti exposed to an ultralow-volume (ULV) aerosol for 15 min and then transferred to clean cages was compared to the mortality of females exposed to the contaminated sentinel exposure cages. ULV aerosol sprays of fenitrothion (50% AI) were dispersed at 180 and 205 ml/min, and ULV aerosol sprays of bendiocarb (18.87% AI) were dispersed at 90 and 120 ml/min in an open field. At 24 h after exposure, the mean percentage of mortality of Ae. aegypti in cages previously exposed to insecticides was 87% at 46 m and 71.8% at 91 m downwind of aerosol generation for the four flow rates. The mean percentage of mortality of Ae. aegypti exposed to the aerosol and then transferred to clean cages was 74.5% at 46 m and 68% at 91 m. Insecticide droplets deposited on exposed cages caused significant mortality to Ae. aegypti. Data show that sentinel mosquitoes used in insecticide bioassays should be transferred into clean holding containers as soon as possible after exposure to prevent biasing of results from insect contact with insecticide deposited on cage walls and screens. PMID- 2566688 TI - Semirigid ureteroscopy: a new genre. AB - The semirigid ureteroscope has a diameter of 7.2F for the distal 11 cm. and gradually enlarges to 11.9F at the proximal portion. We inserted the device in the ureter of 50 consecutive patients without the need for ureteral dilation. The instrument has 2, 0.025-inch channels (2.1F) for wires (0.025), irrigation or 250 mu. laser fibers. Flexible optical fibers and a metal sheath allow for handling as with a rigid instrument, with 2 inches of flexure at the tip without loss of vision. The instrument was inserted successfully in 26 of 27 female patients (lower ureter in 13 of 13, mid ureter in 4 of 4 and upper ureter in 9 of 10), and 14 of 23 male patients (lower ureter in 8 of 10, mid ureter in 4 of 8 and upper ureter in 2 of 5). The semirigid ureteroscope may be used for laser fragmentation of ureteral calculi in almost all female patients and in some male patients. It may be used in strictured and otherwise impassable ureters, for placement of guide wires in the intramural tunnel and to examine the entire ureter atraumatically in most female and some male patients. The device has been designed specifically for laser lithotripsy, and its development expands the indications and facility with which this technology may be used. PMID- 2566689 TI - Infant testicular prostheses. AB - Traditionally, the parents of young boys with an absent testis are advised to let the child decide whether he wants a testicular prosthesis after he enters puberty. Unfortunately, the decision by an adolescent to undergo prosthesis insertion often is made only after experiencing ridicule and embarrassment about the genital appearance. Accordingly, we have offered to implant an infant size prosthesis in patients with cryptorchidism when inguinal and abdominal exploration demonstrates that the testis is absent. A total of 41 boys less than 5 years old with an absent testis underwent simultaneous insertion of a testicular prosthesis. The only complication was a superficial wound infection that did not necessitate removal of the prosthesis. Two boys report mild scrotal discomfort. In long-term followup 91 per cent of the families rated the cosmetic appearance as "good" and 94 per cent were content with the decision regarding placement of a prosthesis. Although many assume that these boys will desire a larger prosthesis after puberty, this consideration seems to be irrelevant. Longer followup will be necessary to ascertain whether this approach alters the psychosexual development of these patients. We believe that young boys with an absent testis should be considered for simultaneous testicular prosthesis insertion at inguinal exploration or orchiectomy. PMID- 2566690 TI - Orchiopexy of the high undescended testis by division of the spermatic vessels: a critical review of 38 selected transections. AB - The technique used to perform 38 orchiopexies of high undescended testes by spermatic vessel transection in 26 patients is reviewed. Careful attention to detail and selection using the Fowler-Stephens test allowed for an 89 per cent success rate, confirming the efficacy of this procedure in cases selected in this manner. PMID- 2566691 TI - Acetylcholine as a possible neurotransmitter in penile erection. AB - We investigated the erectile response to intracavernous injection of increasing doses of acetylcholine (0.5 to 500 micrograms.) in 10 monkeys. To differentiate between nicotinic (ganglionic) and muscarinic (parasympathetic postganglionic) effects, acetylcholine was likewise administered after 1.6 mg. trimethaphan camsylate and 0.1 mg. atropine, alone or sequentially. Erections were induced by cavernous nerve stimulation before and after atropine. Acetylcholine induced a dose-dependent, triphasic erectile response: a first tumescence phase followed by contraction and a subsequent second phase of tumescence. Atropine reduced but did not abolish the erectile response to acetylcholine: attainment of maximal intracavernous pressure after neurostimulation was both delayed and reduced (mean 25 cm. H2O). Only after combined nicotinic and muscarinic blockade was the erectile response to acetylcholine completely abolished. Histologic staining for acetylcholinesterase in five additional monkeys that had not received acetylcholine showed dense staining within the cavernous erectile tissue and around the cavernous arteries. Our data suggest that acetylcholine is a possible neurotransmitter for penile erection in monkeys. PMID- 2566692 TI - In vivo transfer of an R-plasmid in a urinary tract infection model. PMID- 2566693 TI - In vitro intravesical instillation of anticholinergic, antispasmodic and calcium blocking agents (rabbit whole bladder model). AB - The systemic side effects accompanying oral pharmacotherapy of neurogenic bladder dysfunction present significant drawbacks to this type of therapy. In these studies we investigated the effect of intravesical administration of anticholinergic, antispasmodic and calcium blocking agents on pressure response mediated by field stimulation and bethanechol. We used the rabbit in vitro whole bladder model for these experiments. The bladder from a mature male NZW rabbit was mounted in an organ bath as a whole bladder preparation. After control field stimulation and bethanechol stimulation, 20 ml. of saline containing the specific drug being evaluated was instilled into the bladder. At 30 minute intervals, the responses to field stimulation and bethanechol were determined. Two hours after instillation of 100 microM of each specific drug, the inhibition of the contractile response to bethanechol and field stimulation (as % inhibition) was as follows: oxybutynin (95%/64%), verapamil (85%/81%), atropine (68%/31%), diltiazem (47%/39%), and imipramine (44%/47%). Atropine and oxybutynin suppressed the contractile response of the bladder to bethanechol to a much greater extent than that to field stimulation, while verapamil, diltiazem and imipramine suppressed the contractile response to bethanechol and field stimulation to approximately the same extent. Two hours after drug instillation, the intravesical solution was washed out and replaced with saline, but the recovery of the bladder contraction was slow and incomplete. The results of this study suggest that the use of self-intravesical instillation to suppress bladder contractility should be a good therapeutic approach for patients with neurogenic bladder, especially those who are already managed by intermittent catheterization. PMID- 2566694 TI - Technical principles of direct innominate artery revascularization: a comparison of endarterectomy and bypass grafts. AB - Occlusive lesions of the innominate artery that require operation occur infrequently. Direct repair has been performed with low morbidity and mortality. There is debate over the best method of direct reconstruction. Twenty-six patients undergoing transsternal innominate artery repair over a 12-year period were reviewed to determine if either grafting or endarterectomy was superior and what technical factors might be responsible for success or failure. Most of the patients were women. Twenty-four of the patients had atherosclerotic lesions, whereas the other two had Takayasu's arteritis. Either neurologic or right upper extremity symptoms were present in 24 patients. Sixteen patients had grafting, and 10 underwent endarterectomy. There was one death. There were no strokes or transient ischemic attacks. Three patients experienced recurrence of their symptoms; all had failures of reconstruction. The use of a bifurcated graft in one patient was probably responsible for one recurrence of symptoms. Single limb grafts with added side arms are probably preferable to bifurcated grafts. Innominate artery grafting and innominate endarterectomy are equally effective, although grafting is applicable to more patients. Direct transsternal repair is the procedure of choice to correct innominate occlusive disease in patients who are good candidates for correction. PMID- 2566695 TI - Marijuana and benzodiazepines in patients receiving methadone treatment. PMID- 2566696 TI - [Trend in molecular biochemical studies on neurological disorders]. PMID- 2566697 TI - [Hemorrhagic fever with renal syndrome (HFRS) in Japan]. PMID- 2566698 TI - [Classification of HFRS viruses]. PMID- 2566700 TI - [The synthesis, storage and secretion of catecholamines]. PMID- 2566699 TI - [Investigation of nucleoprotein fixation method in immunohistology]. PMID- 2566701 TI - Binding characteristics of 3H-CGP12177 to beta-adrenoceptors in rat myocardial membranes. AB - The present study was designed to examine the selectivity of 3H-CGP-12177 (4-(3-t butylamino-2-hydroxypropoxy)-[5,7-3H]benzimidazole-2-one hydrochloride) for beta 1- and beta 2-adrenergic receptors by the Scatchard and the displacement analysis. The plots of specific binding obtained from the Scatchard analysis using 3H-CGP12177 for the rat myocardium membrane were uniphasic when the non specific binding was determined by the use of 10 microM I-propranolol, and the Kd and Bmax values were 408.53 +/- 67.20 pM and 12.27 +/- 0.83 fmoles/mg protein, respectively. On the other hand, two binding sites were observed in the displacement curve when I-metoprolol was used as a competitor. The existence of these two binding sites implied the selectivity of 3H-CGP12177 to beta adrenoceptors because 3H-CGP12177 was 1.8-fold more selective towards beta 1 adrenoceptors than beta 2-adrenoceptors. In addition, these two binding sites could be regarded as beta 1- and beta 2-adrenergic receptors from the evaluation of the binding characteristics beta-adrenoceptors by the displacement analysis using beta-selective antagonists. Thus, 3H-CGP12177, a hydrophilic radioligand, was useful for the binding assay of beta-adrenoceptors in rat myocardial membranes. PMID- 2566702 TI - Effect of isofloxythepin, a novel neuroleptic, on hippocampal stimulation-induced wet-dog shaking in the rat. AB - (+/-)Isofloxythepin (0.32-3.2 mg/kg, i.p.) significantly inhibited in a dose dependent manner wet-dog shaking (WDS) induced by electrical stimulation of the rat hippocampus. In addition, both optical isomers of isofloxythepin inhibited WDS, with the (-)-isomer being almost 3 times more potent than the (+)-isomer. Other neuroleptics such as haloperidol, chlorpromazine, zotepine and sulpiride also reduced significantly the number of WDS. The inhibitory potency of haloperidol was comparable to that of (+/-)isofloxythepin, which was approximately 3 times more potent than that of chlorpromazine or zotepine. Sulpiride suppressed significantly WDS only at the high dose of 100 mg/kg. None of the drugs affected hippocampal afterdischarge. Inhibition of WDS produced by (+/-) isofloxythepin or haloperidol was antagonized by pretreatment with a dopamine receptor agonist, lisuride. The present results indicate that isofloxythepin shares with other neuroleptics an inhibitory effect on WDS; dopaminergic blocking action appears to be important in the inhibition of WDS induced by hippocampal stimulation. PMID- 2566703 TI - 2-amino-5-phosphonovaleric acid blocks induction of an epileptiform discharge following a brief hypoxic episode in the hippocampal slices prepared from dietary Mg-deficient mouse. AB - The effects of a brief hypoxic episode on synaptic activity in the CA1 region of the hippocampus were studied using the in vitro slice prepared from dietary Mg deficient mouse. After 5 min of hypoxia, the synaptically evoked population spike was increased in amplitude and developed to an epileptiform discharge. The induction of the epileptiform discharge following hypoxia was prevented in the presence of DL-2-amino-5-phosphonovaleric acid (APV), a N-methyl-D-aspartate (NMDA) receptor antagonist, at a concentration of 30-50 microM. The post-hypoxic epileptiform discharge, however, was reduced but not blocked completely by the application of APV (concentration up to 100 microM). These results show that a brief hypoxic episode induces APV-sensitive epileptiform activity in the CA1 region of dietary Mg-deficient mouse, suggesting the involvement of NMDA receptors in post-hypoxic changes of synaptic transmission in the hippocampus. PMID- 2566705 TI - [Impression of the 7th Yokohama regional meeting of public health nursing]. PMID- 2566704 TI - Effects of isofloxythepin on central and peripheral histamine systems. AB - The effects of isofloxythepin, a dibenzo[b,f]thiepin derivative, on the central and peripheral histamine systems were compared with those of chlorpromazine and haloperidol. The three drugs examined all inhibited both the histamine-induced contraction of guinea pig ileum and the specific [3H]mepyramine binding to guinea pig brain membranes in a dose-dependent manner. The effectiveness in inhibiting these reactions was in the order of: chlorpromazine greater than isofloxythepin greater than haloperidol. The histamine-induced relaxation of rat uterus, which is mediated by H2-receptors, was not affected by isofloxythepin. The effect of isofloxythepin on the pargyline-induced accumulation of tele-methylhistamine in the mouse brain was indicative of a decrease in histamine turnover, whereas chlorpromazine and haloperidol were devoid of such effects. Isofloxythepin inhibited both the lethal effect of histamine injected i.v. in mice and histamine induced edema in rat hind paws far more strongly than chlorpromazine or haloperidol did. These results show that isofloxythepin is a neuroleptic with H1 antagonist properties, which are intermediate in potency between those of chlorpromazine and haloperidol, and also it may have an inhibitory action on histamine turnover in the brain. Protection against the lethal effect of histamine and the inhibition of histamine edema by isofloxythepin may largely be due to mechanisms other than the blocking of H1-receptors. PMID- 2566706 TI - [Ways of individualizing use of anti-anginal preparations]. PMID- 2566707 TI - [Local anesthetic effect and subjective tolerance of 2% carteolol and 0.6% metipranolol on healthy eyes]. AB - In a placebo-controlled, randomized, prospective, parallel, double-blind trial, corneal sensitivity was studied in 35 people with healthy eyes before and 1, 3, 6, 10, 15, and 30 minutes after application of carteolol 2% (20 eyes) and metipranolol 0.6% (20 eyes). Oxybuprocain 0.4% (local anesthetic, 10 eyes) served as a positive control substance, NaCl 0.9% (20 eyes) as a negative one (placebo). The test subjects assessed subjective tolerance of the test drugs on a four-step scale. Carteolol 2% did not decrease corneal sensitivity. There was no statistically significant difference between this group and the placebo group. Metipranolol 0.6% led to a decrease in corneal sensitivity of up to 14 mg (median), lasting up to 15 minutes. The decrease brought about by metipranolol 0.6% is about one-half of that caused by betaxolol 0.5%, but greater than that caused by timolol 0.5%. Oxybuprocain 0.4% causes corneal sensitivity to decrease below the measuring range of Cochet and Bonnet's esthesiometer (200 mg) for about 10 minutes. All but three eyes had returned to their initial corneal sensitivity values after 30 minutes. NaCl 0.9% eye drops do not decrease corneal sensitivity. On the contrary, a gradual increase in sensitivity (exercise effect) was found, which differed significantly from the initial value as of the 10th minute after drug application. These results confirm that it is necessary to check corneal sensitivity during topical beta-blocker therapy. "Responders," "sensitives," contact lens wearers, and patients with decreased corneal sensitivity (e.g., patients suffering from open-angle glaucoma with disk excavations, after cataract extraction or corneal transplantation) need frequent supervision.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566708 TI - [Diagnosis of cardiac lesions in the generalized form of meningococcal infection]. AB - Activity of the enzymes indicating heart affection in the acute period and in disease dynamics was increased in 97 patients with generalized forms of meningococcal infection. Signs of cytolysis, according to the data of enzymatic blood examination, were revealed to the utmost degree in patients with typical complications of meningococcal infection--brain edema, which persisted for 10 days of the disease. Herpetic infection joins twice as often in patients with brain edema and is accompanied by a pronounced cytolytic component in the myocardium and liver. Infectious cardiopathy as compared to other complications is characterized by a longer period of hyperenzymemia and the increment of the enzyme activity is greater in the acute period. PMID- 2566709 TI - [Tasks of the reorganization of public health in the light of the decisions of the 19th All-Union Conference of the CPSU and the resolutions of the All-Union Congress of Physicians]. PMID- 2566710 TI - [Errors in the diagnosis of hemorrhagic fever with nephrotic syndrome in surgical practice]. PMID- 2566711 TI - [Pathogenesis of water-electrolyte imbalance in acute renal failure in patients with hemorrhagic fever with nephrotic syndrome]. AB - Changes in blood plasma content of hormones which are observed in the different periods of hemorrhagic fever and the attendant renal syndrome are directed to the maintenance of significantly deranged water-electrolyte homeostasis. Adequate secretion of vasopressin and aldosterone in response to the changes in sodium concentration and plasma osmolality point to the absence of significant functional disorders of the corresponding glands. Pronounced hypernatremia in fatal cases is evidence of the deranged processes of osmoregulation associated primarily with kidney areactivity to vasopressin and prognostically is an unfavourable sign. The presence of pituitary necrosis in deceased subjects does not rule out the role of vasopressin deficiency in the pathogenesis of pronounced hypernatremia. PMID- 2566712 TI - Takayasu's arteritis. PMID- 2566713 TI - Atrial fibrillation after coronary artery bypass grafting. Is it a disorder of the elderly? AB - A total of 1666 patients undergoing isolated coronary artery bypass were studied for the occurrence of postoperative atrial fibrillation. Possible associations of this arrhythmia with various preoperative, intraoperative, and postoperative factors were studied by univariate (chi 2 and t tests) and multivariate (logistic regression) analyses. The overall incidence of postoperative atrial fibrillation was 28.4%, with the major occurrence 2 days after the operation. Both univariate and multivariate studies indicated the patient's age to be the dominant factor promoting postoperative atrial fibrillation, with an increasing prevalence in older patients (p = 0.0001). Multivariate analysis showed that postoperative beta blocker therapy conveyed considerable protection against postoperative atrial fibrillation (p = 0.001) but was less effective in the older patients. Men were more prone to this arrhythmia (p = 0.02). Although these associations appeared significant, the logistic model proved to be a poor predictor of postoperative atrial fibrillation, which suggests that other factors not studied, or mere chance, may also be responsible. PMID- 2566714 TI - Double antigen localization of two catecholamine enzymes and GABA in amacrine cells of the rat retina in semi-thin sections. AB - A double immunohistochemical labeling technique was developed with two immunoperoxidase reactions performed before and after embedding in epoxy resin using two different chromogens: diaminobenzidine (yellowish brown color) and diaminobenzidine + nickel ammonium sulfate (black color). The two catecholamine enzyme immunoreactivities (phenylethanolamine-N-methyltransferase, and tyrosine hydroxylase) were not found in the same cells, while gamma-aminobutyric acid (GABA) immunoreactivity was observed in large tyrosine-hydroxylase immunoreactive cells, but not in phenylethanolamine-N-methyltransferase immunoreactive cells. PMID- 2566715 TI - Intrinsic and synaptic mechanisms of hypothalamic neurons studied with slice and explant preparations. AB - The use of slice and explant preparations has allowed major advances in our understanding of the membrane physiology of mammalian hypothalamic neurons. This article will review intracellular electrophysiological studies of neurons in or immediately surrounding the supraoptic and paraventricular nuclei. Considerable information is now available on the intrinsic membrane mechanisms that control action potential generation and burst firing in magnocellular neuroendocrine cells (MNCs) within these nuclei. Neurons surrounding the paraventricular nucleus have different electrical properties than the MNCs, including low-threshold Ca2+ spikes and pronounced anomalous rectification. Bicuculline and kynurenic acid strongly depress fast IPSPs and EPSPs in MNCs, thus suggesting that inhibitory and excitatory amino acids mediate fast synaptic transmission in the hypothalamus. The effects of neuromodulators, such as noradrenaline and opioid peptides, have also been examined. Noradrenaline excites supraoptic neurons and leads to phasic firing through an alpha-1 mechanism and decreased K+-conductance. Opioid peptides act directly on mu-receptors to hyperpolarize about half of the neurons through an increased K+-conductance. In conclusion, using the magnocellular neuroendocrine system as a model, in vitro slice and explant preparations have allowed the characterization of electrophysiological properties, the identification of neurotransmitters for synaptic events, and studies on the mechanism of action of neuromodulators. PMID- 2566716 TI - [Transplantation of the arm to the contralateral upper arm stump]. AB - Authors performed at a severe injury of the upper limb the transposition of the left hand and lower arm on the stump of the right upper arm. 10 months have passed since the operation. The present state is described. PMID- 2566717 TI - [Early experience with the use of the Halo device in the treatment of damage to the cervical spine]. AB - Authors report on their experiences gained in 5 cases with the Halo device. The method of treatment is described. On the basis of a literary overview the field of indication, the advantages and the disadvantages of the method are described. In the assessment of the method it is stressed that according to their opinion this is the best conservative method of treatment, and the results compete with that of the operative treatment. PMID- 2566718 TI - [Management of hip fractures using a dynamic hip screw]. AB - Authors describe the history of the dynamic hip screw and analyse the hip fracture. The operative technique of the DHS is described in details. In an analysis of the material of the Wilhelminenspital it is stressed that in the last years the DHS became dominant in the treatment of hip fractures. The treatment of four different types of fractures with DHS is demonstrated. The method is compared with other methods suitable for the treatment of hip fractures and it is stated that the less complications can be expected from the use of the DHS. On the basis of this comparison it is shown that the DHS is worthy to occupy a decisive place in the treatment of hip fractures. PMID- 2566719 TI - [Conservative treatment of suppurative hematogenic arthritis of the hip joint]. AB - Authors collected from the 82 suppurations of the hip, during the 12 years activity of their Department, the cases treated from some reason conservatively. They had 21 such cases, 17 of them males. The average age of the patients was 38 years and the haematogenous way of the infection could be proven in 15 cases. The scheme of the treatment: very long.lasting giving of broad spectra antibiotics, temporary immobilization with extension, long-lasting relief of weight bearing, if necessary hip spica. The results are assessed critically. In this the tendency of recurrence of the disease, and that the longest follow-up was 10 years, in average however only 3.6 years, play a role. At closing of the material 12 patients stated to be free of complaints, the hip movements were completely retained in 4.9 cases are demonstrated in details and the "strategic" questions of the conservative treatment of the suppurations of the hip are discussed. PMID- 2566720 TI - [Use of a periosteal flap in the surgical management of rupture of the acromioclavicular joint capsule and ligament. Preliminary report]. AB - Authors use in the treatment of acromioclavicular joint dislocations to increase the safety of the ligament sutures a periosteal flap, gained from the clavicle. The use of the periosteal flap to increase the strength of the acromioclavicular ligament sutures is described. In case of fixing the joint with a tension band the suspension of the cerclage on a screw for an easier removal is suggested. PMID- 2566722 TI - [Pathomechanism of calcium phosphate arthropathy based on a case]. AB - Authors describe the case of a 43 years old male patient with complaints and restriction of motions since years, localized on the talocrural joint. In the background of the disease, responding inadequately to the therapy, calcium pyrophosphate depositions were found in the joint tissues. As the pathomechanism of the disease is even presently not cleared authors summarize our present knowledge referring to this problem. PMID- 2566721 TI - [Grade III open fracture of the femoral diaphysis in a polytraumatized child]. AB - Authors describe the case of a 7.5 years old girl run over and with polytrauma, whose bilateral fracture of the femoral diaphysis was treated with osteosynthesis. The operative indication was the III. degree open fracture on one side. Anatomical reduction was performed on both sides, with DC plate on the closed side and with fixateur externe after insertion of adaptation screws on the open one. The missing soft tissues were substituted with delayed split thickness mashgraft flaps. The injuries healed with good function. If the operative treatment of the femoral diaphysis is founded the lamellar osteosynthesis or the fixateur externe method is suggested. PMID- 2566723 TI - [Shoulder dislocation associated with humeral fracture on the same side (unstable shoulder joint)]. AB - The joint instability observed at the dislocation of the shoulder and the humeral fracture on the same side are described. The idea of "unstable shoulder joint" is used and the operative indication of the shoulder dislocation is outlined. PMID- 2566724 TI - [A rare case of simultaneous bilateral femoral neck fracture]. AB - In connection with a simultaneous bilateral femoral neck fracture in old age author underlines its rare occurrence, even in the international literature. He thinks the conservation of the femoral head even in old age especially significant, in the choice of the operation however the mental state of the elderly must also be taken into consideration. In bilateral femoral neck fracture, on one side by all means, a hemiarthroplasty of sure weight bearing stability is thought to be necessary. PMID- 2566726 TI - Leukaemia Research Fund international research symposium on cytotoxic drug resistance in leukemia and other malignancies. PMID- 2566725 TI - Myeloproliferative disorders: usefulness of X-linked probes in diagnosis. AB - Myeloproliferative disorders are neoplasms of the pluripotent hematopoietic stem cell. Accurate diagnosis and distinction from reactive processes can be difficult therein with cytogenetic analysis only being useful in a minority of patients. Use of X-linked restriction fragment length polymorphism and methylation analysis has enabled clonal analysis to be performed in up to 50% of females, significantly increasing the proportion of analyzable patients over methods dependent on glucose-6-phosphate dehydrogenase heterozygosity. Using hypoxanthine phosphoribosyl transferase and phosphoglycerate kinase probes, we have demonstrated monoclonality of peripheral blood leukocytes in three females with myeloproliferative disorders who had uninformative chromosomal analysis. This technique greatly enhances the diagnosis of early myeloproliferative disorder. PMID- 2566727 TI - [Type 1 fimbriae, P fimbriae and X adhesins in Escherichia strains causing pyelonephritis, cystitis and recurrent urinary infections]. AB - The presence of type 1 fimbriae, P fimbriae and X adhesins in 30 strains of Escherichia coli producing pyelonephritis, 30 strains producing cystitis, and 30 strains producing recurrent urinary infections are compared. All the strains were collected from adult women without risk factors for infections of the urinary tract. The presence of P fimbriae was 73% in strains producing pyelonephritis, 40% in strains producing cystitis and 20% in strains producing recurrent urinary infections; the presence of X adhesins was 3%, 0, and 17% respectively. There were, in consequence, significant differences between the groups studied; however, such differences were not observed with respect to the presence of type 1 fimbriae, which was 97%, 97% and 90%, respectively. We conclude that P fimbriae act as a factor of greater virulence in the strains of E. coli producing pyelonephritis and X adhesins are found almost exclusively in the strains producing recurrent urinary infections. PMID- 2566728 TI - Hepatic imaging and advanced endoscopic techniques. AB - Diseases of the liver and biliary tract can be diagnosed and potentially treated by a variety of radiologic modalities and endoscopic techniques. The imaging modalities of ultrasonography, computed tomography, nuclear scintigraphy and magnetic resonance are emphasized in this article. The current status of endoscopic retrograde cholangiopancreatography, sphincterotomy, biliary endoprostheses, cholangioscopy, endoscopic ultrasound, and laser lithotripsy is discussed. PMID- 2566729 TI - The pharmacology of histamine H2-receptor antagonists. AB - The structure of the first histamine H2-receptor antagonist, burimamide was described in 1972. Since then, numerous compounds with diverse chemical structures have been shown to possess H2-receptor antagonist activity. Most of these compounds comprise an aromatic ring and polar group linked by a flexible chemical chain. Currently five H2-receptor antagonists are available for therapeutic use. In vitro studies have confirmed the competitive nature of the interaction between agonist and antagonist for these compounds and a comparison of the pA2 values generated shows a rank order of potency cimetidine less than nizatidine = ranitidine less than or equal to roxatidine less than famotidine. The duration of action of these agents is largely dependent upon their pharmacokinetic properties; with the possible exception of roxatidine, all have plasma elimination half-lives in the range 2-4 hours. Competitive antagonists with a high degree of selectivity for the H2-receptor and with a longer duration of action are currently being investigated, e.g., sufotidine. In addition to competitive agents, several non-competitive H2-antagonists, e.g., loxtidine, have been described. These have a prolonged duration of action. Relative potency varies between animal species and pharmacological models studied but the results of animal pharmacological studies have generally provided a good indication of the likely effects in man. Studies of 24-hour intragastric acidity represent the most physiological model available in man for evaluating the effects of different antisecretory drug regimens. Comparative studies with ranitidine 150 mg b.i.d. and cimetidine 400 mg b.i.d. have shown a reduction in 24-hour acidity of 65% and 30%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566730 TI - Clinical importance of the new H2-antagonists. AB - The first H2-receptor antagonist, cimetidine, has been succeeded by ranitidine, and more recently nizatidine and famotidine. Others will doubtless follow. The pharmacodynamic differences between cimetidine and ranitidine, in terms of potency, are not obviously translated into therapeutic differences. Some studies have shown that the increased potency of standard doses of ranitidine affords an advantage, in terms of ulcer healing and relapse, over cimetidine. The therapeutic effects of these drugs can be predicted from their pharmacodynamic profiles and in the doses recommended differences between the newest agents and ranitidine would not be expected. It is, thus, difficult to define a specific clinical role for the newer drugs on the grounds of efficacy. Their importance may relate to the provision of encouragement for future drug development. PMID- 2566731 TI - [The 4th international meeting in Paris: current aspects of the biology and treatment of chronic lymphatic leukemia]. PMID- 2566732 TI - [What to do with a patient who demands treatment based on false premises?]. PMID- 2566733 TI - A cAMP binding abnormality in psoriasis. AB - In 34 psoriatic patients with various cutaneous manifestations (psoriasis vulgaris, erythroderma psoriaticum, guttate psoriasis), the ability of the RI regulatory subunit of cAMP-dependent protein kinase (PKA) to bind a cAMP analogue (8-azido [32P] cAMP) in erythrocyte membranes was significantly lower than that in 19 normal subjects (mean [SEM] 565 [35] vs 930 [35] fmol/mg protein). This enzyme defect was not found in patients with other forms of dermatitis that can be confused with psoriasis or with other inflammatory diseases. There was a significant negative correlation between the severity of the disease as expressed by the psoriatic area and severity index score and the binding of the cAMP analogue to PKA. A long-term study showed that oral retinoid treatment of psoriatic patients resulted in a correction of the binding defect. Unaffected members of psoriatic families had significantly lower than normal binding of cAMP to PKA (773 [60] fmol/mg protein). This study shows for the first time that in psoriasis a biochemical defect expressed in erythrocytes correlates with the severity of the disease as well as its clinical evolution. These results will be useful in clinical management of psoriatic disease for the choice and follow-up of retinoid therapy. PMID- 2566734 TI - Bacteriotherapy for chronic relapsing Clostridium difficile diarrhoea in six patients. AB - Six patients with chronic relapsing diarrhoea caused by Clostridium difficile were treated with rectal instillation of homologous faeces (one patient) or a mixture of ten different facultatively aerobic and anaerobic bacteria diluted in sterile saline (five patients). The mixture led to a prompt loss of Cl difficile and its toxin from the stools and to bowel colonisation by Bacteroides sp, which had not been present in pre-treatment stool samples. Strains of Escherichia coli, Cl bifermentans, and Peptostreptococcus productus in the mixture inhibited the in vitro growth of Cl difficile, which in turn inhibited the growth of Bacteroides ovatus, Bacteroides vulgatus, and Bacteroides thetaiotaomicron. The finding that Bacteroides sp had been absent during the patients' illness but was present after recovery suggests that the absence of Bacteroides sp may result in chronic relapsing Cl difficile diarrhoea, and that its presence may prevent colonisation by Cl difficile. PMID- 2566735 TI - Randomised, controlled trial of faecal occult blood screening for colorectal cancer. Results for first 107,349 subjects. AB - To assess the effectiveness of screening by faecal occult blood tests, 107,349 people without symptoms of colorectal disease identified from general practitioner records have been randomly allocated to test and control groups. 53,464 test subjects were invited to carry out the screening test; 27,651 (53%) of the 52,258 who received the tests did so. Further investigation of the 618 (2.3%) with positive tests showed 63 cancers (52% stage A) and 367 adenomas (266 subjects). Rescreening of subjects with negative results every 2 years (9510 first rescreen, 3639 second) has shown a significant fall in the rate of positive results (1.7% of 7344; 0.3% of 2906). Cancers have also been diagnosed in 20 subjects presenting in the interval between a negative test and rescreening, and in 83 non-responders. The incidence of cancer in the control group (123 subjects; 10.6% stage A) was 0.72 per 1000 person-years. Cancers detected by screening were at a less advanced pathological stage, but it is too early to show any effect of screening on mortality from colorectal cancer. PMID- 2566737 TI - Campylobacter pylori and duodenal ulcers: the gastrin link. AB - The possibility that Campylobacter pylori (CP) in the gastric antrum stimulates gastrin release in duodenal ulcer (DU) disease was examined in 31 patients. The 25 patients with antral colonisation with CP had higher basal and meal-stimulated plasma gastrin concentrations, and higher peak acid output (PAO), than did the 6 without CP in the autumn. PMID- 2566736 TI - Comparison of endoscopic polidocanol injection and YAG laser therapy for bleeding peptic ulcers. AB - 392 patients were examined by endoscopy for acute upper gastrointestinal bleeding; 140 had ulcers containing an actively bleeding visible vessel or a non bleeding visible vessel and were enrolled in a randomised trial of three endoscopic methods of haemostasis--adrenaline (1/10(4] alone, adrenaline plus polidocanol 1%, and adrenaline followed by yttrium-aluminium-garnet (YAG) laser photocoagulation. For patients with non-bleeding visible vessels sham treatment was significantly less effective in achieving haemostasis (8 of 20 patients) than were adrenaline plus polidocanol (18 of 20; p = 0.002) and adrenaline plus laser (16 of 20; p = 0.012). All three treatments significantly reduced total transfusion needs compared with sham treatment. For the whole group of patients, adrenaline plus polidocanol was significantly more effective than adrenaline alone in achieving permanent haemostasis; adrenaline plus laser was also more effective than adrenaline alone, but not significantly so. The efficacy of the three treatments was enhanced by repeated application on recurrence of bleeding. Since injection therapy with adrenaline and polidocanol was at least as effective as adrenaline plus laser therapy, it should be preferred over laser therapy because it is cheaper, easier to use, and perhaps also safer. PMID- 2566738 TI - Epstein-Barr virus silver anniversary. PMID- 2566739 TI - Mitral valve prolapse. PMID- 2566740 TI - Write for your patient. PMID- 2566741 TI - Stroboscopic examination of the larynx. PMID- 2566742 TI - Sensitive thyrotropin measurements: utility and futility. PMID- 2566743 TI - Dietary cholesterol and ischaemic heart disease. AB - In a cohort of 1824 middle-aged men followed for 25 years, intake of dietary cholesterol was associated with risk of death from ischaemic heart disease, from other cardiovascular diseases combined, from all cardiovascular diseases combined, and from all causes combined. The relative hazard of death from all cardiovascular diseases combined, associated with the difference between the mean of the first and fifth quintiles of cholesterol intake (a difference of 184 mg cholesterol/1000 kcal intake) was 1.46 (95% confidence interval 1.10-1.94) after adjustment for age, intake of other dietary lipids, and other coronary risk factors (including serum cholesterol). When stratified into three groups according to serum cholesterol (less than 220 mg/dl, 220-259 mg/dl, and 260 mg/dl or above), the corresponding relative hazards were 1.58, 1.50, and 1.41, respectively. These results are further evidence for the concepts that dietary cholesterol is atherogenic in man, and that the effect is partly independent of total serum cholesterol. They reinforce the recommendation that intake of dietary cholesterol should be low in people without overt hyperlipidaemia as well as those with raised serum cholesterol. PMID- 2566744 TI - Platelet-derived growth factor. PMID- 2566745 TI - Diagnostic value of an immunoassay to detect anti Campylobacter pylori antibodies in non-ulcer dyspepsia. AB - An enzyme-linked immunosorbent assay (ELISA) for detection of IgG antibodies against Campylobacter pylori was used to examine sera from 70 patients with non ulcer dyspepsia. 48 patients had C pylori associated gastritis according to culture or histology; mean optical density (OD) of the ELISA was significantly higher than that for the 22 patients with normal antral mucosa and absence of C pylori. Positive and negative predictive values for campylobacter-associated gastritis were 100% above OD 2.10 and below OD 1.00, respectively. Serology might replace endoscopy in the diagnosis of campylobacter-associated gastritis. PMID- 2566746 TI - Oocyte donation in 61 patients. AB - Oocyte donation was performed by gamete intrafallopian transfer in 61 women, 34 of whom were amenorrhoeic. The mean age of the donors was 32 years (range 23-38). Seventy-five treatment cycles gave 29 clinical pregnancies, of which 21 reached term, 3 continue, and 5 were lost (3 miscarriages, 1 tubal, and 1 cervical). 11 (38%) of the women who became pregnant were over 42 years old. When more than four oocytes were transferred, many of the pregnancies were multiple. 8 (38%) of the pregnancies that came to term were complicated by pre-eclamptic toxaemia. When oocytes are obtained from young women, the fertility potential is high irrespective of the recipient's age. PMID- 2566747 TI - Should all pregnant women be screened for gestational glucose intolerance? AB - Although gestational glucose intolerance is associated with the remote development of diabetes mellitus, the risk to the mother during the index pregnancy and the risk to her fetus remain uncertain. Nevertheless, universal screening for gestational glucose intolerance has many strong advocates. The scientific data supporting a universal screening programme--showing that treatment of gestational glucose intolerance does more good than harm--are limited. Until the evidence can be extended beyond that on infant birthweight, a more restrained approach than universal screening may be appropriate. PMID- 2566748 TI - Prevention of torture and inhuman or degrading treatment: medical implications of a new European convention. AB - A new European convention creates a mechanism for the prevention of torture and inhuman or degrading treatments of detained people through visits by outside, independent teams with unlimited access to places of detention. The convention has important implications for the medical profession: firstly, visits to psychiatric hospitals will be included and, in particular, to secure facilities, where the risk of human rights abuses is well established; and, secondly, the adequacy and ethics of medical care in prisons will be a key issue in assessing the protection of prisoners' human rights. The convention should be welcomed by the medical profession as a stimulus to the improvement of medical care for detained people. PMID- 2566749 TI - World no tobacco day: a challenge for women's health. PMID- 2566750 TI - Fenoterol and asthma mortality. PMID- 2566751 TI - Fenoterol and serum potassium. PMID- 2566752 TI - Mortality and colorectal surgery. PMID- 2566753 TI - Dirofilariasis of the breast. PMID- 2566754 TI - Lewis negative genotype and breast cancer risk. PMID- 2566755 TI - Enterovirus infection and cardiomyopathy in Cameroon. PMID- 2566756 TI - Outcome of childhood minimal-change disease. PMID- 2566757 TI - Plasmapheresis for late-stage trypanosomiasis. PMID- 2566758 TI - Thrombolytic therapy for superior vena cava syndrome. PMID- 2566759 TI - Echocardiography in diagnosis of aortic dissection. PMID- 2566760 TI - Candidate gene approach to type IIa hypercholesterolaemia. PMID- 2566762 TI - Molecular genetics of amyloid neuropathy in Europe. PMID- 2566761 TI - Detection of asymptomatic coronary artery disease in familial hypercholesterolaemia. PMID- 2566763 TI - Varicella zoster virus and recurrent aphthous ulceration. PMID- 2566764 TI - British diplomas and limited registration. PMID- 2566765 TI - Differential capitation rates. PMID- 2566766 TI - Scurvy and anaemia in refugees. PMID- 2566767 TI - Adoptive immunotherapy. PMID- 2566768 TI - Anti-HIV activity of castanospermine analogues. PMID- 2566769 TI - Augmentation of in-vitro HIV replication in peripheral blood mononuclear cells of AIDS and ARC patients by tumour necrosis factor. PMID- 2566770 TI - Mercury poisoning from dental amalgam through a direct nose-brain transport. PMID- 2566771 TI - Effect of suramin in a patient with adrenocortical carcinoma. PMID- 2566772 TI - Greenhouse effect. PMID- 2566773 TI - Anaesthetic agents and the ozone layer. PMID- 2566774 TI - Fatal pneumonia after glandular fever and rubella. PMID- 2566776 TI - Human organ transplants bill. PMID- 2566775 TI - Phenytoin and fasting. PMID- 2566777 TI - Plasma immunoreactive endothelin in uraemia. PMID- 2566778 TI - The Canadian American Ticlopidine Study (CATS) in thromboembolic stroke. AB - The Canadian American Ticlopidine Study (CATS) is a randomised, double-blind, placebo-controlled trial to assess the effect of ticlopidine (250 mg twice daily) in reducing the rate of subsequent occurrence of stroke, myocardial infarction, or vascular death in patients who have had a recent thromboembolic stroke. Twenty five centres entered 1072 patients into the study between 1 week and 4 months after their qualifying stroke. The patients were treated and followed for up to 3 years (mean 24 months). In the efficacy analysis, the event rate per year for stroke, myocardial infarction or vascular death, considered together, was 15.3% in the placebo group and 10.8% in the ticlopidine group, representing a relative risk reduction with ticlopidine of 30.2% (95% confidence interval 7.5-48.3%; p = 0.006). Ticlopidine was beneficial for both men and women (relative risk reductions 28.1%, p = 0.037, and 34.2%, p = 0.045, respectively). Analysis by intention-to-treat gave a smaller estimate of risk reduction (23.3%, p = 0.020) for stroke, myocardial infarction, or vascular death. Adverse experiences associated with ticlopidine included neutropenia (severe in about 1% of cases) and skin rash and diarrhoea (severe in 2% of cases each); all were reversible. This study provides evidence of a beneficial effect of ticlopidine in both men and women with a recent thromboembolic stroke. PMID- 2566779 TI - Beneficial effects of growth hormone treatment in GH-deficient adults. AB - A double-blind, placebo-controlled, crossover study on the effects of 4 months' growth hormone (GH) treatment was carried out in 22 GH-deficient adults (8 women, 14 men; mean [SEM] age 23.8 [1.2] years). 1 patient was withdrawn because of oedema. Mean total body weight of the other 21 did not change, whereas mean muscle volume of the thigh, estimated by computerised tomography (CT), was significantly higher after GH than after placebo (70.0 [3.7] vs 66.3 [3.1] ml/0.8 cm cross-sectional slice). The mean adipose tissue volume of the thigh and subscapular skinfold thickness fell significantly during GH treatment. Growth hormone caused a small increase in the isometric strength of the quadriceps muscles and a significant rise in exercise capacity (60.8 [7.2] vs 54.2 [6.6] kJ). The heart rate both at rest and after maximum exercise was low during the placebo period and increased significantly during GH treatment. Blood pressure and echocardiographic wall mass of the left ventricle did not change during the study. Growth hormone increased both mean glomerular filtration rate and renal plasma flow from a subnormal level on placebo to a level comparable with that of an age-matched control group. The filtration fraction did not change. Urinary albumin excretion was in the low normal range and was not affected by GH treatment. Finally, GH treatment normalised mean circulating levels of insulin like growth factor 1 (IGF-1), which were low after the placebo period (96 [9] micrograms/l placebo; 224 [28] micrograms/l GH). These findings suggest that GH, in a conventional replacement dose, has several potentially beneficial effects in GH-deficient adults and therefore encourage future long-term trials. PMID- 2566780 TI - Relation between infection with a subtype of HPV16 and cervical neoplasia. AB - A variant (HPV16b) of the previously described human papillomavirus (HPV)16 (now called HPV16a) has been identified by polymerase chain reaction assays. Compared with HPV16a, HPV16b has a 21bp deletion and several point mutations within the upstream regulatory region. All cervical carcinoma samples studied contained only HPV16a. HPV16b was found only rarely in cervical neoplasia but was common in the normal population. PMID- 2566782 TI - Tin-protoporphyrin and long wave length ultraviolet light in treatment of psoriasis. AB - To assess the effects of tin (Sn)-protoporphyrin (a synthetic haem analogue) in conjunction with long wave length ultraviolet light (UVA) radiation in psoriasis 10 patients, 9 of whom were substantially or completely unresponsive to other forms of therapy, received 2.0 mumol/kg bodyweight of Sn-protoporphyrin for 1 day followed by UVA light treatment for 21 days. The average starting UVA dose was 5.6 (SD 2.0) J/cm2, and the average cumulative UVA dose was 98.3 (35.1) J/cm2. Severity of psoriatic plaques, scored clinically on a scale of 0-3 for erythema, scaling, and induration, fell from a mean score of 7.9 at the start of the study to 3.6 at the end. Psoriatic lesions were improved in all patients and the effect was striking in some. The responses lasted throughout the three weeks of the study and no deleterious side-effects of the treatment were noted. Clinical follow-up for three months showed no rebound in disease activity. Sn protoporphyrin with conventional UVA light may be useful in the treatment of psoriasis. PMID- 2566781 TI - Prevention of transfusion-acquired cytomegalovirus infection in infants by blood filtration to remove leucocytes. Neonatal Cytomegalovirus Infection Study Group. AB - A multicentre, controlled trial was carried out to determine whether removal of leucocytes from blood by means of 'Imugard IG500' (Terumo) filters would prevent transfusion-acquired cytomegalovirus (CMV) infection in newborn infants. 72 infants whose mothers were seronegative and who received some seropositive blood were followed for 6 months for evidence of CMV infection. There were no significant differences between the groups who received filtered and unfiltered blood in median gestation, birthweight, or amount of seropositive blood received (median volume 32.5 ml and 34.5 ml, respectively). 9 (21%) of the 42 infants who received unfiltered blood and none of the 30 who received filtered blood were infected with CMV. All infected infants weighed less than 1500 g at birth; they represented 31% of very low birthweight (VLBW) infants at risk of CMV infection. None of 24 VLBW infants who received filtered seropositive blood was infected. 1 infected infant died and 5 had clinical features consistent with CMV infection. The results show that transfusion-acquired CMV infection is preventable by filtration of blood through a leucocyte filter. This method has advantages over other methods of removing leucocytes or the use of only seronegative blood for newborn infants. PMID- 2566783 TI - Diabetic honeymoon: prolonged at a price? PMID- 2566784 TI - Colon cancer: molecular analysis marches on. PMID- 2566785 TI - Surveillance cultures in neutropenia . PMID- 2566786 TI - Skin surface microscopy: anything new under the sun? PMID- 2566787 TI - Hyperkalaemia--silent and deadly. PMID- 2566788 TI - Cigarette smoking, polyunsaturated fats, linoleic acid, and coronary heart disease. PMID- 2566789 TI - Epidermal growth factor and related molecules. PMID- 2566791 TI - Maternal pyrexia associated with the use of epidural analgesia in labour. AB - To establish the effect of pain relief on maternal temperature during labour forty patients who went into spontaneous labour with a single fetus, had a normal temperature (less than 37.5 degrees C), and had no clinical evidence of infection were investigated prospectively. They were divided into two comparable groups- one receiving pethidine and the other epidural analgesia. Both groups had much the same temperatures at the beginning of labour and before any analgesic administration. The mean temperature in the pethidine group remained constant during labour, whereas in the epidural analgesia group it showed a significant rise after only 6 hours of labour. This rise was not related to any clinical evidence of infection. Patients receiving epidural analgesia during labour are at increased risk of developing pyrexia. This pyrexia may be the result of vascular and thermoregulatory modifications induced by epidural analgesia. PMID- 2566790 TI - Trial of prednisolone for prevention of melarsoprol-induced encephalopathy in gambiense sleeping sickness. AB - In a prospective randomised trial, 620 patients who had Trypanosoma brucei gambiense trypanosomiasis with central nervous system involvement were treated either with prednisolone plus melarsoprol or with melarsoprol only. 598 patients were evaluable: morbidity and death associated with melarsoprol-induced encephalopathy was reduced in patients who were given prednisolone. The two groups did not differ either in the incidence of other complications of melarsoprol therapy or in relapse rate after melarsoprol therapy. The cost of prednisolone would be outweighed by savings on the treatment of encephalopathies in such patients. PMID- 2566792 TI - The "Help Your Patient Stop" initiative. Evaluation of smoking prevalence and dissemination of WHO/UICC guidelines in UK general practice. AB - The World Health Organisation and the International Agency against Cancer in 1988 published joint guidelines on smoking cessation for primary health care teams. A booklet entitled Help Your Patient Stop was produced in the United Kingdom as a model for the international dissemination of these guidelines. This booklet was sent to UK general practitioners by post; about 4 weeks later, a random sample of 5000 were asked to complete a postal questionnaire about the booklet and their smoking habits. The response rate was 75%. About half (50.5%) remembered receiving the booklet, 27.7% had read it, and only 8.8% could write down any of the three essential activities in smoking cessation which the booklet was intended to promote and which were printed in bold letters on the inside back cover. Although the booklet itself might be an adequate model for other countries, unless dissemination and marketing of the information it contains can be improved, its achievement will be limited. However, the survey did have one optimistic feature: only 13.5% of general practitioners reported that they smoke; and only a third of those who gave full details of their smoking habit smoke cigarettes. PMID- 2566793 TI - Oral contraceptives and breast cancer. PMID- 2566794 TI - Long-term effects of methyl isocyanate. PMID- 2566796 TI - Minimal residual leukaemia in chronic myeloid leukaemia patients after T-cell depleted bone-marrow transplantation. PMID- 2566795 TI - Difficulty in detection of hernia and hydrocele in cryptorchidism. PMID- 2566797 TI - Adenosine deaminase and tuberculous peritonitis. PMID- 2566798 TI - Leukoaraiosis. PMID- 2566799 TI - IgG endomysial antibodies in IgA-deficient patient with coeliac disease. PMID- 2566800 TI - Interaction of fluconazole with cyclosporin. PMID- 2566801 TI - Apomorphine test for dopaminergic responsiveness. PMID- 2566802 TI - Heart failure responding to octreotide in patient with acromegaly. PMID- 2566803 TI - Interferon and hepatitis B vaccine in haemodialysis patients. PMID- 2566804 TI - Peripheral benzodiazepine receptors and treatment of porphyria. PMID- 2566805 TI - Failure to detect cytomegalovirus-DNA in IgA nephropathy by in-situ hybridisation. PMID- 2566806 TI - Histiocytic medullary reticulosis, a lethal form of primary EBV infection. PMID- 2566807 TI - Interactions between haemopoietic growth factors. PMID- 2566808 TI - Mitochondrial cytopathy and neurodegenerative diseases of old age. PMID- 2566809 TI - Skin cancer among atom bomb survivors. PMID- 2566810 TI - Cardiac arrhythmia suppression trial and flecainide. PMID- 2566811 TI - Localised prostatic cancer. PMID- 2566812 TI - HER2/neu amplification and comedo type breast carcinoma. PMID- 2566813 TI - Mitochondrial complex I deficiency in Parkinson's disease. PMID- 2566814 TI - Ascariasis. PMID- 2566815 TI - Sonography in diagnosis of fascioliasis. PMID- 2566816 TI - Fat embolism and patent foramen ovale. PMID- 2566817 TI - Rapid diagnosis of malaria. PMID- 2566818 TI - Acute renal failure after nebulised pentamidine. PMID- 2566819 TI - Immunological treatment for multiple sclerosis. PMID- 2566820 TI - Modifying hysterectomy rates. PMID- 2566821 TI - Bovine cartilage and marrow extract. PMID- 2566822 TI - Third World and European community. PMID- 2566823 TI - Sterilisation of a mentally incapable woman. PMID- 2566824 TI - Randomised comparison of chloramphenicol, ampicillin, cefotaxime, and ceftriaxone for childhood bacterial meningitis. Finnish Study Group. AB - In a multicentre study, 220 consecutive cases of bacterial meningitis in children older than 3 months were randomised to treatment with chloramphenicol, ampicillin (initially with chloramphenicol), cefotaxime, or ceftriaxone. The drugs were given in four equal daily doses for 7 days, except ceftriaxone which was given only once daily. 200 cases could be assessed; the causative organisms were Haemophilus influenzae type b (Hib) in 146; meningococci (Mnc) in 32; pneumococci (Pnc) in 13; and other or unknown in 9. In patients with Hib meningitis, sterilisation of the cerebrospinal fluid occurred most rapidly with ceftriaxone. Otherwise, in terms of overall clinical recovery, normalisation of laboratory indices, clinically significant adverse reactions, toxic effects, sequelae, and mortality rate, the treatment groups were very similar. However, there were 4 bacteriological failures, all in the chloramphenicol group. Also, the treatment was extended or changed in more cases in the chloramphenicol group than in the other groups. Chloramphenicol was thus inferior to the other three antimicrobials. Ampicillin is a good and cheap alternative, but there are difficulties with resistance. Easy administration tempts the use of ceftriaxone rather than cefotaxime but it causes diarrhoea. A 7-day course of ampicillin, cefotaxime, or ceftriaxone is sufficient in Hib, Mnc, or Pnc meningitis. PMID- 2566825 TI - Effects of intradermal gamma-interferon in cutaneous leishmaniasis. AB - The clinical and immunological evolution of lesions in cutaneous leishmaniasis was assessed after treatment with human recombinant gamma interferon (rIFN gamma). 3 weeks after rIFN-gamma treatment of lesions due to Leishmania braziliensis guyanensis, 12/13 had become smaller compared with 6/13 control lesions; only 4 treated lesions were free of parasites. 9 of 13 L tropica lesions treated with rIFN-gamma resolved completely within 4-8 weeks of treatment. An acute inflammatory reaction around treated lesions was more common in lesions due to L tropica. There were no other local or systemic adverse reactions. Histological and immunohistochemical studies indicate that local application of rIFN-gamma enhances cell-mediated immune responses and thus promotes healing of cutaneous leishmaniasis. PMID- 2566826 TI - Linkage between immunoglobulin E responses underlying asthma and rhinitis and chromosome 11q. AB - Family studies of IgE responses to common inhaled antigens have suggested dominant inheritance of atopy. Molecular genetic linkage analysis was used to confirm this proposal. In seven families the transmission of atopy was linked, with a maximum lod score of 5.58, to a DNA polymorphism defined by p lambda MS.51, which confirms dominant inheritance and assigns the gene locus to chromosome 11. PMID- 2566827 TI - Controlled trial of haem arginate in acute hepatic porphyria. AB - A double-blind study comparing placebo and haem arginate was conducted in 12 patients with acute intermittent porphyria. 2 days after admission in attack patients were randomised to receive intravenous haem arginate 3 mg/kg per 24 h for 4 days or placebo. 9 patients were readmitted with a further attack and were given the alternative treatment. Before randomisation the paired attacks were of similar severity with respect to urinary porphobilinogen (PBG) excretion and clinical manifestations. With haem arginate the median PBG excretion of the 9 patients with two attacks (normal range 0-16 mumol per 24 h) fell significantly from 332 mumol per 24 h (range 137-722) on admission to a median lowest level of 40 (range 22-105). On placebo, median PBG excretion was 382 (range 196-542) on admission, falling to 235 (range 128-427). Median duration of admission after the start of treatment was 11 days (range 2-28) for placebo and 8 days (3-26) for haem arginate. Median total analgesic requirement between the start of treatment and discharge was 8150 mg pethidine equivalents (range 0-17,650) with placebo versus 6425 (range 50-20,650) with haem arginate. Phlebitis occurred in 5 patients on haem arginate and in 2 on placebo. Haem arginate effectively reduces porphyrin precursor overproduction in the acute porphyric attack but this reduction is not accompanied by striking resolution of the clinical manifestations of the attack. PMID- 2566828 TI - Contribution of aminoacid catabolism to propionate production in methylmalonic acidaemia. AB - The rates of propionate production and aminoacid catabolism in 5 children with methylmalonic acidaemia were measured by use of stable isotope techniques. Total propionate production was 55-186 mumol/kg per h, to which the maximum contribution of protein catabolism was 10-35 mumol/kg per h (5-40%). These findings indicate important sources of propionate other than protein catabolism, which may account for the limited efficacy of dietary protein restriction in treatment of methylmalonic acidaemia. PMID- 2566829 TI - Salt and blood pressure: the next chapter. PMID- 2566830 TI - UK obstetrics and the new health service. PMID- 2566831 TI - Cerebrospinal fluid shunt infections. PMID- 2566832 TI - Risk factors for back trouble. PMID- 2566833 TI - Solving problems, taking decisions. PMID- 2566834 TI - Urgent endoscopic drainage for acute suppurative cholangitis. AB - 105 patients with acute calculous cholangitis who did not respond to conservative management underwent urgent endoscopic drainage of the biliary system at a mean of 1.5 days after admission. Treatment was successful in 102 (97%) patients. 3 of the patients in whom drainage was not successful underwent emergency surgery, with 1 death. 3 patients died of uncontrolled sepsis despite successful endoscopic drainage. 1 patient died of a stroke. The overall mortality was 4.7%. Among those in shock 2 out of 4 drained after 72 h died, compared with 3 out of 38 drained before 72 h. There were no deaths in the group without shock irrespective of the timing of drainage. PMID- 2566835 TI - Arrhythmia in heart failure: role of mechanically induced changes in electrophysiology. AB - Various mechanisms have been suggested to explain the high prevalence of ventricular arrhythmia in patients with heart failure, but as yet there is no unifying theory. There is growing evidence that changes in myocardial mechanical properties may directly alter cardiac electrophysiology by a process of mechanoelectric feedback. Moreover, when changes in cardiac loading similar to those seen in heart failure are produced experimentally in normal heart, there is a greater tendency to arrhythmogenesis. The intimate relation between changes in mechanical function and arrhythmia in heart failure could account for the lack of effect of most conventional antiarrhythmic drugs on arrhythmogenesis, and the beneficial effect of peripheral vasodilators. This paper argues that mechanically induced changes in electrophysiology are very important in the development of arrhythmia in cardiac failure; there may be no need to implicate other mechanisms, such as relative ischaemia, metabolic changes, or changes in sympathetic tone. PMID- 2566836 TI - Peptide regulatory factors in embryonic development. PMID- 2566837 TI - An initiative on vesicovaginal fistula. PMID- 2566838 TI - Torture of Turkish Kurds. PMID- 2566839 TI - Bradyarrhythmia with University of Wisconsin preservation solution. PMID- 2566840 TI - Chechum (poison sap tree) toxin, a potent activator of protein kinase C. PMID- 2566841 TI - Antecollis in parkinsonism. PMID- 2566842 TI - Selection for unrelated donor bone marrow transplantation. PMID- 2566843 TI - Desmopressin and Jehovah's Witness. PMID- 2566844 TI - Pathogenesis of progressive multiple sclerosis. PMID- 2566845 TI - Ondansetron with and without dexamethasone to treat chemotherapy-induced emesis. PMID- 2566846 TI - Delayed human chorionic gonadotropin administration for in-vitro fertilisation. PMID- 2566847 TI - HTLV-I and blood donors. PMID- 2566848 TI - Uncertainties about AIDS and HIV. PMID- 2566849 TI - Sensitivity of HIV screening tests. PMID- 2566850 TI - Rapid decrease in melatonin production during successful treatment of delayed puberty. PMID- 2566851 TI - In-vivo 31P magnetic resonance spectroscopy for monitoring treatment response in breast cancer. PMID- 2566852 TI - Pyruvate kinase deficiency and delayed clinical response to recombinant human erythropoietin treatment. PMID- 2566853 TI - Ifosfamide-induced Fanconi syndrome. PMID- 2566854 TI - Angina pectoris, adenosine, and theophylline. PMID- 2566855 TI - Fetal brain tissue. PMID- 2566856 TI - Promotion of medicines. PMID- 2566857 TI - UK health education. PMID- 2566858 TI - Abortion USA. PMID- 2566859 TI - Medication during Ramadan. PMID- 2566860 TI - Latent colorectal cancer found at necropsy. PMID- 2566861 TI - Adrenaline response to hypoglycaemia and insulin species. PMID- 2566862 TI - Neonatal hypoglycaemia. PMID- 2566863 TI - Interferon in lungs. PMID- 2566864 TI - Prevention of acute graft-versus-host disease by monoclonal antibody to interleukin-2 receptor. PMID- 2566865 TI - Elimination by interferon-alpha of malignant clone in chronic myeloid leukaemia. PMID- 2566866 TI - Guillain-Barre syndrome and human diploid cell rabies vaccine. PMID- 2566867 TI - Does caffeine withdrawal contribute to postanaesthetic morbidity? PMID- 2566869 TI - Ascariasis. PMID- 2566868 TI - Bronchoconstriction and pentamidine. PMID- 2566870 TI - Handling echovirus 11 outbreaks. PMID- 2566871 TI - Kussmaul's sign: an artifact? PMID- 2566872 TI - Skin cancer and the ozone layer. PMID- 2566873 TI - Osteoarthritis and cartilage collagen genes. PMID- 2566874 TI - Morbidity of very-low-birthweight infants. PMID- 2566875 TI - Empirical treatment of pneumococcal pneumonia in Spain. PMID- 2566876 TI - Neonatal tetanus (St Kilda, 19th century) PMID- 2566878 TI - AIDS prevention strategies. PMID- 2566877 TI - Benzodiazepine sedation and allegations of sexual assault. PMID- 2566879 TI - The effect of cetirizine on early allergic response. AB - A double blind, placebo-controlled, cross-over study was performed to determine the effect of cetirizine, an H1 antihistamine, on the immediate nasal allergic response. Ten persons underwent nasal challenge with antigen after premedication with 20 mg of cetirizine or placebo QD for 2 days. The response was monitored by counting the number of sneezes and by measuring the levels of histamine, prostaglandin D2, leukotriene C4, albumin, and TAME-esterase activity in recovered nasal lavages. The results showed a significant reduction in sneezing and in the amounts of recovered albumin, TAME-esterase activity, and leukotriene C4 but no reduction in the amounts of recovered histamine and prostaglandin D2. These results suggest that cetirizine does not inhibit mast cell activation but inhibits the consequences of the released histamine on H1 receptors: sneezing and increased vascular permeability. The results further suggest that mast cell release of histamine is the direct result of antigen stimulation, as opposed to reflex activation, and that other cells in addition to mast cells generate leukotrienes during the early allergic response. PMID- 2566880 TI - Systemic vasculitis: a temporal bone histopathologic study. AB - Systemic vasculitis includes a broad spectrum of disorders that may involve blood vessels of any size in any organ system. Systemic vasculitis is associated with immunopathogenic mechanisms. Sixteen temporal bones from eight persons were studied to determine histopathologic changes that occur in systemic vasculitis. Three persons had Wegener's granulomatosis, two had polyarteritis nodosa, and three had systemic lupus erythematosus. Otitis media was seen in 15 ears, with ten ears showing chronic middle ear changes and two showing fibrotic inner ear changes. In Wegener's granulomatosis, granulation tissue was observed around the eustachian tube and protympanum, and in polyarteritis nodosa, inflammatory cell infiltrate and thickened blood vessels were observed around the facial nerve. Although sensorineural hearing loss has been described clinically in systemic lupus erythematosus, the present report describes findings in temporal bones, including severe fibrosis and new bone formation throughout the inner ear. PMID- 2566881 TI - Dependence on extracellular potassium of the positive inotropic response to St 587, a selective alpha-1 adrenoceptor agonist, in Zucker rat heart ventricle. AB - The effects of St 587, a selective alpha-1 adrenoceptor agonist, were investigated in non obese and obese Zucker rat heart ventricles. In both groups, the numbers and affinity constants for alpha-1 adrenoceptors were found to be similar. At 4 or 10 mM [K]o, St 587 failed to increase the developed tension whereas at 14 mM [K]o, St 587 significantly increased it in both groups of rats. This effect was reversed by prazosin; St 587 also increased action potential duration at 14 mM [K]o. [K]o is thus important for the occurrence of the inotropic effect of St 587 in 12 week-old Zucker rats, either non obese or obese with reduced beta-adrenoceptor responsiveness. This suggests the participation of phosphoinositide metabolism in the mechanism of St 587 inotropic effect in the rat. PMID- 2566882 TI - Localization of dynorphin-induced neurotoxicity in rat spinal cord. AB - Intrathecally injected dynorphin A (1-13) in rats results in a reversible hindlimb paralysis and an irreversible loss of the tail-flick reflex. Histologic examination of the spinal cords of dynorphin treated rats demonstrated dead and/or dying neurons predominately localized in the central area which approximates Rexed lamina VII and X. In this area a maximum effect of the dynorphin-induced neurotoxicity is evident. Thus, the dynorphin-induced neuron death is suggestive of an anatomical selectivity. PMID- 2566883 TI - Chronic lesions differentially decrease tyrosine hydroxylase messenger RNA in dopaminergic neurons of the substantia nigra. AB - Long-term effects of lesions were analyzed in terms of gene expression. Nine months after unilateral 6-hydroxydopamine (6-OHDA) lesions of the substantia nigra pars compacta (s. nigra), the remaining dopaminergic (DAergic) neurons (tyrosine hydroxylase (TH) cells determined by immunocytochemistry (ICC] on the lesioned side were atrophic with smaller nucleoli. By in situ hybridization, the DAergic neurons on the lesioned side had a 50% smaller TH-mRNA concentration than on the contralateral non-lesioned side. However, beta-tubulin mRNA concentration in DAergic neurons was unaffected by the lesion. The lesions did not alter TH mRNA concentration in the contralateral non-lesioned side by comparison with unoperated controls. We propose that chronic lesions have long-term effects on gene expression because of damage sustained during compensatory hyperactivity after the lesion, or because of decreased trophic support from other neurons. PMID- 2566884 TI - Amygdaloid kindling of rats increases preprosomatostatin mRNA and somatostatin without affecting glutamic acid decarboxylase (GAD) mRNA or GAD. AB - The levels of preprosomatostatin (preproSS) mRNA, somatostatin-like immunoactivity (SS-LI) (also known as somatotropin-release inhibitory factor, or SRIF), glutamic acid decarboxylase (GAD) activity and GAD mRNA were determined in several brain regions of amygdaloid-kindled rats. SS mRNA and SS increased in the cortex and striatum, while only SS increased in the hippocampus. No changes were detected in either GAD activity or GAD mRNA in any brain region. The data suggest that somatostatin may be one of the factors involved in the chain of events leading to kindled seizures. PMID- 2566885 TI - [Tranquilizers]. PMID- 2566886 TI - Effects of stretching and disuse on amino acids in muscles of rat hind limbs. AB - Effects of stretching on muscle amino acids were tested in unloaded soleus by casting the foot in dorsiflexion on one limb of tail-casted, hindquarter suspended rats. For comparison with unloading, amino acids also were measured in shortened extensor digitorum longus (EDL) in the same casted limb and in denervated leg muscles. Concentrations of tyrosine and glutamate were lower, while aspartate, ammonia, and the ratio of glutamine to glutamate were greater in the stretched than in the freely moving, unloaded soleus, but stretched did not differ from weight-bearing, control muscle. Therefore, stretching the soleus muscle prevented changes in certain amino acids due to unloading. Aspartate, ammonia, glutamine, and the ratio of glutamine to glutamate were lower in the shortened EDL than in the freely moving muscle of the contralateral limb, or in the control muscle. When denervated, these leg muscles also showed lower aspartate, ammonia, and ratio of glutamine to glutamate relative to innervated muscles. Since muscle shortening or denervation produced amino acid changes that mimicked the effects of unloading on the soleus, these responses must reflect the effect of muscle disuse. These data suggested that lower ammonia might cause the lower ratio of glutamine to glutamate with disuse. Because the fresh muscle energy charge, one factor which controls AMP deaminase, generally was not affected by disuse, altered deamination of glutamate via glutamate dehydrogenase may explain the variations in muscle ammonia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566887 TI - Effect of repeated ingestion of aspartame-sweetened beverage on plasma amino acid, blood methanol, and blood formate concentrations in normal adults. AB - Aspartame (APM) is a widely used dipeptide sweetener (L-aspartyl-L-phenylalanine methyl ester). It has been suggested that excessive use of APM might elevate plasma aspartate, phenylalanine, and/or methanol concentrations to levels that are potentially harmful. Six normal young adults ingested eight successive servings of unsweetened and APM-sweetened beverage at one-hour intervals in a balanced crossover design. In one part, the beverage was not sweetened. In the other, each serving of beverage provided 600 mg APM, a dose equivalent to the amount provided by 36 oz of APM-sweetened diet beverage. Plasma aspartate concentration was not significantly increased after ingestion of unsweetened or APM-sweetened beverage. Similarly, ingestion of the unsweetened beverage had no significant effect on plasma phenylalanine concentration. However, ingestion of APM-sweetened beverage significantly increased plasma phenylalanine levels 1.41 to 2.35 mumol/dL above baseline 30 minutes after ingestion. Plasma phenylalanine values reached a steady state after administration of four to five servings and did not exceed normal postprandial values at any time. Blood methanol and formate concentrations remained within normal limits. The data indicate ready metabolism of APM when administered at levels that may be ingested by normal individuals who are heavy users of diet beverages. PMID- 2566888 TI - Effects of cimaterol, a beta-adrenergic agonist, on lipid metabolism in rats. AB - Rats fed a high carbohydrate diet containing 10 or 100 ppm cimaterol for 4 weeks gained 41% to 59% less fat and 70% to 76% more protein than controls, with no major changes in either energy gain or efficiency of energy retention. Effects of cimaterol on lipid metabolism in these rats were assessed. Cimaterol stimulated lipolysis in vivo and in vitro, but failed to influence rates of de novo fatty acid synthesis in either liver or white adipose tissue. Activities of fatty acid synthetase and malic enzyme in these tissues were also unaffected by cimaterol. Cimaterol administered in vivo failed to affect lipoprotein lipase activity in white adipose tissue, but elevated enzyme activity 67% to 75% in the extensor digitorium longus muscle. Lipoprotein lipase activity in the extensor digitorum longus muscle was also elevated by 66% during a 2 hour incubation with 1 mmol/L cimaterol. We conclude that cimaterol selectively stimulates both lipolysis in white adipose tissue and lipoprotein lipase activity in skeletal muscle, to direct energy away from adipose tissue deposition toward skeletal muscle accretion. PMID- 2566889 TI - Somatostatin effect in postprandial hypoglycemia. AB - The effect of the administration of oral glucose with or without a simultaneous intravenous somatostatin infusion on blood glucose, immunoreactive insulin, C peptide, and glucagon levels in seven patients with idiopathic postprandial hypoglycemia was studied. Oral glucose alone induced an excessive insulin response and hypoglycemia, whereas a slight suppression in glucagon levels without any increase at the hypoglycemic nadir was observed. The simultaneous administration of somatostatin significantly reduced the insulin response and induced a slower rise of blood glucose; no hypoglycemia developed. Only minor variations in glucagon were observed with respect to the basal test. A rebound in insulin, C-peptide, and glucagon levels was observed at the end of the somatostatin infusion. These data show that somatostatin can suppress glucose induced hypoglycemia in these subjects, thus suggesting that its long-acting analogues might be worth a therapeutic trial in severe idiopathic postprandial hypoglycemia. PMID- 2566890 TI - Use of microdialysis in neuroendocrinology. PMID- 2566891 TI - Primary thyrotropin-releasing hormone-degrading enzymes. PMID- 2566892 TI - Quantification of the mass of tyrosine monooxygenase in the median eminence and superior cervical ganglion. AB - A procedure is described that enables one to quantify the mass of TH in a fraction of the ME and SCG of rats. This procedure is specific and sensitive. It should be possible to study the biosynthetic activity of catecholaminergic neurons as a function of the mass of TH in such cells. PMID- 2566893 TI - Passive immunoneutralization: a method for studying the regulation of basal and pulsatile hormone secretion. AB - The combination of passive immunoneutralization of an endogenous factor with frequent sampling techniques to delineate the pattern of secretion represents a powerful tool for dissecting the contribution of that factor to normal and/or abnormal pituitary function. Passive immunoneutralization offers all of the advantages of other known means of abolishing endogenous factors. It has the specificity and the selectivity of antagonists while retaining the potential for replacement of biological activity that is possible with organ removal or selective brain cuts or lesions. The only major drawback to this approach is the potential difficulty in obtaining the appropriate reagents. The inherent flexibility of the model, however, increases the probability of obtaining useful data with those reagents that are available. At present, the number of studies using immunoneutralization is relatively small, but the knowledge gained from these studies has been substantial. As the number of immunoneutralization and replacement studies increases in the future, the usefulness of this powerful approach will be extended to its full potential. PMID- 2566894 TI - Methods for the study of somatostatin. AB - It is clear from the above that there are a number of methods for study of SRIF release. From the standpoint of convenience, the in vitro static incubation of ME is the most practical technique at the present time. Using this preparation SRIF release has been found to be modified by a number of neurotransmitters and peptides, and studies on the mechanism of release of the peptide have been initiated. There is no doubt that such studies should be complemented by perifusion studies, by studies involving larger pieces of the hypothalamus which encompass the entire somatostatinergic neuron, and by in vivo studies to determine the correlation of in vivo and in vitro release. Among the in vivo techniques which have been utilized, the push-pull cannula technique employing cannulae implanted in hypothalamus or anterior pituitary gland offers the most promise. A summary of the effects of some neurotransmitters and neuropeptides on hypothalamic SRIF secretion is reported in Table. I. PMID- 2566895 TI - Application of fast atom bombardment mass spectrometry to posttranslational modifications of neuropeptides. AB - FABMS is a powerful and sensitive analytical technique capable of providing structural information unattainable by standard methods of peptide analysis. Many posttranslational modifications are undetectable by other routine analytical methods. In addition, FABMS is capable of providing information regarding posttranslational modifications at levels of peptide comparable to those required for other methods of analysis (10-1000 pmol). FABMS has had the effect on protein structure analysis that structure determination of any neuropeptide might now be considered incomplete without some form of mass spectrometric analysis. Much of the recent explosive increase in the use of mass spectrometry for solving problems in peptide structure analysis can be traced to improvements in methods capable of producing molecular ions from nonvolatile species. With the development of these methods, it can be expected that refinements of existing methods and new ionization methods will continue to increase the mass range and sensitivity available for peptide structure determination. For a brief review of other mass spectrometric methods applicable to peptides, see Delgass and Cooks. PMID- 2566897 TI - The acquired immunodeficiency syndrome and mosquitoes. PMID- 2566896 TI - Comparison of virulence between two strains of Rattus serotype hemorrhagic fever with renal syndrome (HFRS) virus in newborn rats. AB - Two strains of hemorrhagic fever with renal syndrome (HFRS) virus from Rattus, SR 11 and KI-262, showed virtually identical antigenicity but differed from prototype strain Hantaan 76-118 (Apodemus origin) in a neutralization test. Wistar newborn rats inoculated intraperitoneally (i.p.) with SR-11, which was isolated from a laboratory rat associated with an outbreak of HFRS, developed clinical signs such as ataxia and limb paralysis and died at about 18 days after inoculation. The LD50 of SR-11 in 1-day-old rats was 10(1.2) focus-forming units (FFU). In contrast, the animals inoculated i.p. or intracerebrally with 10(4) FFU of KI-262, which was from a wild rat in a dumping-ground area--an enzootic focus where no human cases have been recorded--did not show any significant clinical signs. The susceptibility of rats to SR-11 fatal infection was age-dependent. Virus titers in brains, lungs, kidneys, and livers of the rats inoculated with SR 11 were significantly higher than those in the same organs of the animals infected with KI-262. Necrosis of neurons in the brain tissue occurred in the rats infected with SR-11, while it was mild in the animals infected with KI-262. PMID- 2566898 TI - Fourth International Conference on the acquired immunodeficiency syndrome. PMID- 2566899 TI - [The course and prognosis of ulcerative colitis]. PMID- 2566900 TI - Postherpetic neuralgia. AB - Postherpetic pain persisting 1 month or longer occurs in only a small percentage of all patients with herpes zoster. In most patients, PHN tends to diminish with time. The incidence is, however, directly related to age. Any therapeutic claim for prophylaxis or treatment of PHN has to be evaluated with these observations in mind. There is some information about the pathologic features and a concept of the pathogenesis can be suggested. There is evidence for an imbalance in fiber input (reduced large, inhibitory fibers, and intact or increased small, excitatory fibers) to an abnormal dorsal horn that may contain hypersensitive neurons. Prevention of PHN remains difficult. There is evidence that systemic steroids exert a preventive effect when employed in the treatment of herpes zoster in the immunocompetent patient. A reasonable regimen is 60 mg of prednisone tapered over 10 to 14 days. One double-blind, controlled study supports the use of amantadine in this situation; this drug is an option in patients for whom steroids are contraindicated, such as those with peptic ulcer, diabetes mellitus or compromised immune function. The dosage of amantadine used in this study was 100 mg twice daily for a month. Although a number of other therapies have been suggested, these remedies remain in need of further, more scientific study. For established PHN, there is firm support for the reduction of pain from severe to mild in two thirds of patients administered low doses of amitriptyline followed by gradual, small increments. In the age group over 65 years, one may use as small a dose as 10 mg with an increase of 10 mg every 5 to 7 days. In those younger than 65, a dose of 25 mg to start is reasonable, with increments of 25 mg. Although unproved, the addition of a phenothiazine, such as fluphenazine, may provide further pain relief. Preliminary studies also indicate that topical capsaicin may be a useful new treatment. Although widely used, there is no good evidence for the use of anticonvulsants alone in this disorder. Studies of local anesthetic sprays with vibration and continuous TENS are uncontrolled, but these modalities may be of some merit. One uncontrolled study reported benefit from epidural steroids. DREZ lesions are a possibility in failed medical cases, but other surgical procedures appear to be of little or no use. Although the measures described here will benefit a number of patients, PHN remains an intractable problem in some cases.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2566901 TI - Painful polyneuropathy. AB - Pain can complicate a diverse group of peripheral neuropathies. A number of potential mechanisms have been proposed, but available data are inadequate to posit any pathogenesis with certainty. In most cases, knowledge of the clinical findings associated with the various syndromes permits an etiologic diagnosis, which, in turn, may suggest a primary therapy. Symptomatic treatment is usually needed, however, and includes a large number of pharmacologic therapies, as well as ancillary physiatric and psychological treatments. PMID- 2566902 TI - Interaction of 6-cyano-7-nitroquinoxaline-2,3-dione with the N-methyl-D-aspartate receptor-associated glycine binding site. AB - The interaction of newly described antagonist of the non-NMDA glutamate receptor 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) with the glycine site of the NMDA receptor complex has been investigated. In whole-cell patch recordings from hippocampal neurons maintained in culture, currents induced by N-methyl-D aspartate (NMDA) were dependent on extracellular glycine. Responses to both NMDA (30 microM) and kainate (20 microM) were reduced by CNQX (10-30 microM). The antagonism by CNQX of NMDA, but not kainate, receptor-mediated responses could be reversed by increasing the concentration of glycine in the external medium. Glycine concentration-response curves constructed in the presence of 30 microM NMDA were shifted to the right by CNQX, suggesting that CNQX was competing with glycine for the glycine binding site. However, even at high concentrations of glycine (300 microM) the maximal NMDA current obtained in the presence of CNQX (10-30 microM) was not restored to control levels. Because CNQX had no effect on responses produced by supramaximal concentrations of NMDA (500 microM) and glycine (300 microM), it is suggested that CNQX also interacts with the NMDA recognition site. The antagonism of currents induced by NMDA was not dependent on the membrane potential, and the rapid onset and offset of the block suggested that there was little or no use dependence. Radioligand binding experiments were performed using [3H]glycine to label the strychnine-insensitive glycine regulatory site of the NMDA receptor complex in guinea pig brain frontal cortex membranes. CNQX displaced [3H]glycine binding in a concentration-dependent manner (IC50 = 5.7 microM). Scatchard analysis of the inhibition showed a decrease in the affinity (increase in Kd) of [3H]glycine binding, but no change in the number of binding sites (Bmax) in the presence of 5 microM CNQX, suggesting a competitive interaction. These data provide evidence that CNQX antagonizes NMDA receptor-mediated responses by competing with glycine for a modulatory site associated with the NMDA receptor complex. Furthermore, the results indicate that CNQX may not be as selective an antagonist for non-NMDA receptors as initially described, although its selectivity will depend on the concentration of the NMDA receptor ligand and may be enhanced by increasing the extracellular concentration of glycine. PMID- 2566903 TI - Characterization of dopamine receptor subtypes by comparative structure-activity relationships: dopaminomimetic activities and solid state conformation of monohydroxy-1,2,3,4,4a,5,10,10a-octahydrobenz[g]quinolines and its implications for a rotamer-based dopamine receptor model. AB - A series of phenolic (cis)- and (trans)-1,2,3,4,4a,5,10,10a octahydrobenz[g]quinolines were investigated in the D1 and D2 dopamine (DA) models, DA-sensitive adenylate cyclase and electrically evoked acetylcholine release, respectively, and were compared with the effects of the corresponding aminotetralins. A similar structure-activity pattern was found at both DA receptor subtypes. The change from the bicyclic to the tricyclic DA analogs resulted in a loss of activity of all beta-rotameric 8-hydroxy derivatives, suggesting the presence of a steric barrier. Derivatives of the alpha-rotameric 6 hydroxy trans series, in contrast to their inactive cis analogs, showed stimulatory effects that increased from N-methyl to N-n-propyl substitution, indicating an interaction with an N-alkyl binding site. The inactivity of the corresponding N-n-butyl derivative ("N-butyl phenomenon") suggests that the N alkyl substituents of this series point toward a "small N-alkyl binding site," which can be differentiated from a "large N-alkyl binding site." An X-ray of the active (-)-enantiomer of (trans)-6-hydroxy-N-n-propyloctahydrobenz[g]quinoline (R)-mandelat e revealed a 4aR, 10aR absolute configuration, corresponding to that of (-)-5-hydroxy-2-(N,N-di-n-propylamino)tetralin. The hydrogen bonding interactions of the axial N+-H proton and the hydroxy group to mandelate anions in the crystal provide a model for a possible drug-receptor interaction. Molecular modeling served to localize the steric barrier and the boundaries of the small N-alkyl binding site, which together form an "extended steric barrier." The results led to the proposal of a refined version of a rotamer-based general DA receptor model, which is supplemented by criteria for the orientation of DA agonists. Its application is demonstrated with apomorphine and ergoline. PMID- 2566904 TI - Two distinct quisqualate receptors regulate Ca2+ homeostasis in hippocampal neurons in vitro. AB - Addition of quisqualate to mouse hippocampal neurons in vitro elicited two types of changes in [Ca2+]i as assessed by fura-2-based microfluorimetry. The first was a transient spike or group of oscillations and the second was a long lasting "plateau" response. The long-lasting response was abolished on removal of either Ca2+ or Na+ from the external medium or by blocking voltage-sensitive Ca2+ channels. Furthermore, the novel glutamate antagonist 6-nitro-7-cyano-quinoxaline 2,3-dione was a competitive inhibitor of this response. In contrast, none of these manipulations abolished the transient [Ca2+]i spike. Transient [Ca2+]i spikes or oscillations could also be produced by the alpha 1-adrenergic agonist phenylephrine. Production of such an alpha 1-response reduced the size of a subsequently elicited quisqualate response. However production of transient [Ca2+]i spikes with caffeine did not alter the size of the quisqualate-induced spike. We conclude that hippocampal neurons possess two different types of quisqualate receptors. The first mediates quisqualate-induced depolarization and the second mediates Ca2+ mobilization from intracellular stores. PMID- 2566905 TI - Consequences of terbium (III) binding on the conformation and enzymatic activity of guinea pig liver transglutaminase. AB - Calcium ions are crucial for expression of transglutaminase activity. Although lanthanides have been reported to substitute for calcium in a variety of protein functions, they did not replace the calcium requirement during transglutaminase activity measurements. Furthermore, lanthanides strongly inhibited purified liver transglutaminase activity using either casein or fibrinogen as substrates. Terbium (III) inhibition of transglutaminase-catalyzed putrescine incorporation into casein was not reversed by the presence of 10-200 fold molar excess of calcium ions (Ki for Tb(III) = 60 microM). Conformational changes in purified liver transglutaminase upon Tb(III) binding were evident from a biphasic effect of Tb(III) on transglutaminase binding to fibrin. Low concentrations of Tb(III) (1 microM to 10 microM) inhibited the binding of transglutaminase to fibrin, whereas higher concentrations (20 microM to 100 microM) promoted binding. Conformational changes in purified liver transglutaminase consequent to Tb(III) binding were also demonstrated by fluorescence spectroscopy due to Forster energy transfer. Fluorescence emission was stable to the presence of 200 mM NaCl and 100 mM CaCl2 only partially quenched emission. Purified liver transglutaminase strongly bound to Tb(III)-Chelating Sepharose beads and binding could not be disrupted by 100 mM CaCl2 solution. Our data suggest that Tb(III)-induced conformational changes in transglutaminase are responsible for the observed effects on enzyme structure and function. The potential applications of Tb(III) transglutaminase interactions in elucidating the structure-function relationships of liver transglutaminase are discussed. PMID- 2566906 TI - Mitogen-induced genes are subject to multiple pathways of regulation in the initial stages of T-cell activation. AB - The delivery of a mitogenic signal to T cells via any one of several cell surface molecules elicits a variety of intracellular responses, some or all of which regulate subsequent gene expression events. The expression of nine novel mitogen induced genes in response to various T-cell-activating agents was examined to evaluate the diversity of pathways which regulate such genes. The relative contribution of distinct secondary signals, individually or together, to mitogen stimulated gene induction and the capability of individual genes to respond to the sometimes divergent signals generated from different cell surface structures is addressed. The activation of T cells with mitogenic monoclonal antibodies directed against the CD2 or CD3 cell surface molecules, or with phytohemagglutinin, induced all nine genes. Thus, stimulation by fully mitogenic agents regardless of cell surface-binding specificity correlated with the expression of all of the genes studied. However, heterogeneous patterns of gene expression, encompassing five regulatory classes, were revealed by the use of phorbol 12-myristate 13-acetate, calcium ionophore, and anti-CD28 monoclonal antibody, agents which mediated only a subset of intracellular events and thus an incomplete mitogenic signal. Interleukin-2 and two novel lymphokines represented one regulatory class that appeared to require unique transcriptional activation signals relative to the other mitogen-induced genes. As demonstrated in the accompanying paper (P. F. Zipfel, S. G. Irving, K. Kelly, and U. Siebenlist, Mol. Cell. Biol. 9:1041-1048, 1989), the immediate transcriptional response of T cells to mitogenic stimulation is quite complex, involving numerous genes beyond those which have been previously described. Furthermore, the discrimination of several regulatory phenotypes among these nine genes suggests that a multiplicity of signaling pathways extends from the cell surface to the level of transcription. PMID- 2566907 TI - p185HER2 monoclonal antibody has antiproliferative effects in vitro and sensitizes human breast tumor cells to tumor necrosis factor. AB - The HER2/c-erbB-2 gene encodes the epidermal growth factor receptorlike human homolog of the rat neu oncogene. Amplification of this gene in primary breast carcinomas has been show to correlate with poor clinical prognosis for certain cancer patients. We show here that a monoclonal antibody directed against the extracellular domain of p185HER2 specifically inhibits the growth of breast tumor derived cell lines overexpressing the HER2/c-erbB-2 gene product and prevents HER2/c-erbB-2-transformed NIH 3T3 cells from forming colonies in soft agar. Furthermore, resistance to the cytotoxic effect of tumor necrosis factor alpha, which has been shown to be a consequence of HER2/c-erbB-2 overexpression, is significantly reduced in the presence of this antibody. PMID- 2566908 TI - Identification of members of the P-glycoprotein multigene family. AB - Overproduction of P-glycoprotein is intimately associated with multidrug resistance. This protein appears to be encoded by a multigene family. Thus, differential expression of different members of this family may contribute to the complexity of the multidrug resistance phenotype. Three lambda genomic clones isolated from a hamster genomic library represent different members of the hamster P-glycoprotein gene family. Using a highly conserved exon probe, we found that the hamster P-glycoprotein gene family consists of three genes. We also found that the P-glycoprotein gene family consists of three genes in mice but has only two genes in humans and rhesus monkeys. The hamster P-glycoprotein genes have similar exon-intron organizations within the 3' region encoding the cytoplasmic domains. We propose that the hamster P-glycoprotein gene family arose from gene duplication. The hamster pgp1 and pgp2 genes appear to be more closely related to each other than either gene is to the pgp3 gene. We speculate that the hamster pgp1 and pgp2 genes arose from a recent gene duplication event and that primates did not undergo this duplication and therefore contain only two P glycoprotein genes. PMID- 2566909 TI - Transcriptional regulation of acetyl coenzyme A carboxylase gene expression by tumor necrosis factor in 30A-5 preadipocytes. AB - Acetyl coenzyme A (acetyl-CoA) carboxylase activity, amount, and mRNA levels increase during the differentiation of 30A-5 preadipocytes to adipocytes. Tumor necrosis factor (TNF) completely prevents this differentiation, with concomitant inhibition of acetyl-CoA carboxylase mRNA accumulation. To investigate the mechanisms by which TNF prevents acetyl-CoA carboxylase mRNA accumulation, we determined the effect of TNF on the transcription rate of the carboxylase gene and the half-life of carboxylase mRNA. Nuclear runoff transcription assays revealed no differences in the number of RNA polymerase molecules actively engaged in transcription of the acetyl-CoA carboxylase gene in preadipocytes, adipocytes, TNF-treated preadipocytes, or at any time during the course of differentiation. However, changes in adipsin, glycerophosphate dehydrogenase, and actin mRNAs, whose levels are also differentiation dependent, can be accounted for in part by changes in the number of polymerase complexes on their respective genes. To determine whether TNF caused a decrease in the stability of carboxylase RNA transcripts, we measured the rate of decay of prelabeled acetyl-CoA carboxylase mRNA. Control and TNF-treated cells showed no difference between the apparent half-lives of acetyl-CoA carboxylase mRNAs (9 h). However, the rate of acetyl-CoA carboxylase mRNA synthesis in vivo was decreased three- to fourfold in the presence of TNF. These data demonstrate that TNF prevents accumulation of acetyl-CoA carboxylase mRNA during preadipocyte differentiation by decreasing the rate of acetyl-CoA carboxylase gene transcription. However, transcriptional control is not due to a change in the number of RNA polymerase complexes actively engaged in carboxylase transcript elongation which could be measured by a number runoff assay. Instead, transcriptional control may be related to the rate at which RNA polymerase traverses the acetyl-CoA carboxylase gene. PMID- 2566910 TI - Role of poly(A) polymerase in the cleavage and polyadenylation of mRNA precursor. AB - To determine the role of poly(A) polymerase in 3'-end processing of mRNA, the effect of purified poly(A) polymerase antibodies on endonucleolytic cleavage and polyadenylation was studied in HeLa nuclear extracts, using adenovirus L3 pre mRNA as the substrate. Both Mg2+- and Mn2+-dependent reactions catalyzing addition of 200 to 250 and 400 to 800 adenylic acid residues, respectively, were inhibited by the antibodies, which suggested that the two reactions were catalyzed by the same enzyme. Anti-poly(A) polymerase antibodies also inhibited the cleavage reaction when the reaction was coupled or chemically uncoupled with polyadenylation. These antibodies also prevented formation of specific complexes between the RNA substrate and components of nuclear extracts during cleavage or polyadenylation, with the concurrent appearance of another, antibody-specific complex. These studies demonstrate that (i) previously characterized poly(A) polymerase is the enzyme responsible for addition of the poly(A) tract at the correct cleavage site and probably for the elongation of poly(A) chains and (ii) the coupling of these two 3'-end processing reactions appears to result from the potential requirement of poly(A) polymerase for the cleavage reaction. The results suggest that the specific endonuclease is associated with poly(A) polymerase in a functional complex. PMID- 2566912 TI - Biochemistry of Parkinson's disease 28 years later: a critical review. PMID- 2566913 TI - A current analysis of behavioral problems in patients with idiopathic Parkinson's disease. PMID- 2566914 TI - Imaging techniques in Parkinson's disease. PMID- 2566911 TI - Sequences downstream of AAUAAA signals affect pre-mRNA cleavage and polyadenylation in vitro both directly and indirectly. AB - To investigate the role of sequences lying downstream of the conserved AAUAAA hexanucleotide in pre-mRNA cleavage and polyadenylation, deletions or substitutions were constructed in polyadenylation signals from simian virus 40 and adenovirus, and their effects were assayed in both crude and fractionated HeLa cell nuclear extracts. As expected, these sequences influenced the efficiency of both cleavage and polyadenylation as well as the accuracy of the cleavage reaction. Sequences near or upstream of the actual site of poly(A) addition appeared to specify a unique cleavage site, since their deletion resulted, in some cases, in heterogeneous cleavage. Furthermore, the sequences that allowed the simian virus 40 late pre-RNA to be cleaved preferentially by partially purified cleavage activity were also those at the cleavage site itself. Interestingly, sequences downstream of the cleavage site interacted with factors not directly involved in catalyzing cleavage and polyadenylation, since the effects of deletions were substantially diminished when partially purified components were used in assays. In addition, these sequences contained elements that could affect 3'-end formation both positively and negatively. PMID- 2566915 TI - Early dopamine agonist therapy in Parkinson's disease. PMID- 2566916 TI - Clonal hematopoiesis demonstrated by X-linked DNA polymorphisms after allogeneic bone marrow transplantation. AB - We analyzed DNA from peripheral-blood and marrow cells from 12 recipients of allogeneic bone marrow transplants to determine whether monoclonal but otherwise normal hematopoiesis occurs in such patients. All patients were being treated for various forms of leukemia or lymphoma. In 10 patients, granulocytes isolated from peripheral-blood samples obtained 28 to 159 days after transplantation were polyclonal. In some, circulating T cells were isolated and also found to be polyclonal. In contrast, two patients had donor-derived monoclonal or oligoclonal hematopoiesis after transplantation. In one, DNA from circulating mononuclear cells obtained 29 days after transplantation revealed a monoclonal pattern on analysis of a restriction-fragment-length polymorphism in the phosphoglycerate kinase gene. In the other, analysis of a restriction-fragment-length polymorphism in the hypoxanthine phosphoribosyltransferase gene suggested the presence of a dominant clone in the granulocytes sampled 36 days after transplantation. When the latter patient was reassessed on day 267, the same clone of donor hematopoietic cells was still predominant and was found to include circulating T cells as well as granulocytes. We conclude that monoclonal hematopoiesis of donor origin may be observed in recipients of allogeneic bone marrow transplants, indicating that stem cells in normal adult human marrow are able to repopulate both lymphoid and myeloid compartments after transplantation. PMID- 2566917 TI - Cyclin synthesis drives the early embryonic cell cycle. AB - We have produced extracts of frog eggs that can perform multiple cell cycles in vitro. Destruction of the endogenous messenger RNA arrests the extracts in interphase. The addition of exogenous cyclin mRNA is sufficient to produce multiple cell cycles. The newly synthesized cyclin protein accumulates during each interphase and is degraded at the end of each mitosis. PMID- 2566918 TI - The role of cyclin synthesis and degradation in the control of maturation promoting factor activity. AB - We show that cyclin plays a pivotal role in the control of mitosis. A proteolysis resistant mutant of cyclin prevents the inactivation of maturation promoting factor and the exit from mitosis both in vivo and in vitro. We have used a fractionated extract to study the activation of MPF by added cyclin protein. PMID- 2566919 TI - CD2-mediated adhesion facilitates T lymphocyte antigen recognition function. AB - The CD2 T lymphocyte-surface glycoprotein serves to mediate adhesion between T lymphocytes and their cognate cellular partners which express the specific ligand LFA-3. In addition, CD2 by itself or in conjunction with T-cell receptor stimulation, transduces signals resulting in T-lymphocyte activation. One or both of these functions seems to be physiologically important, given that certain anti CD2 monoclonal antibodies block T-cell activation and that antigen-responsive memory T cells express a high level of CD2 relative to virgin T cells, which are largely antigen-unresponsive. Nevertheless, the contribution of the individual CD2 functions in T-cell responses has not been independently examined. To this end, human CD2 complementary DNAs encoding an intact LFA-3-binding adhesion domain, but lacking a functional cytoplasmic signal transduction element (CD2trans-), were introduced into an ovalbumin-specific, I-Ad restricted murine T cell hybridoma. The antigen-specific response of T hybridoma cells expressing human CD2trans- protein was enhanced up to 400% when the human LFA-3 ligand was introduced into the I-Ad expressing murine antigen-presenting cells. In contrast, no augmentation was observed if human LFA-3 was absent or expressed on a third party cell lacking the I-Ad restriction element. These results directly demonstrate the functional significance of adhesion events mediated between CD2 on the antigen-responsive T lymphocyte and LFA-3 on the presenting cell in optimizing antigen-specific T-cell activation. PMID- 2566920 TI - AIDS conference: uncertain start in Montreal. PMID- 2566921 TI - On minimum cyclic AMP formation rates associated with positive inotropic effects mediated through beta 1-adrenoceptors in kitten myocardium. Beta 1-specific and non-adrenergic stimulant effects of denopamine. AB - The agonist (-)-denopamine was used as a tool to study relationships between pharmacological effects and adenylate cyclase stimulation mediated through beta 1 adrenoceptors. 1. (-)-Denopamine was a full agonist in kitten papillary muscles (force), kitten left atria (force) and kitten and guinea-pig atria (sinoatrial frequency). (-)-Denopamine was a strong partial agonist in guinea-pig tracheae (relaxation). None of these effects was influenced by blockade of beta 2 adrenoceptors. beta 1-Adrenoceptors mediated all effects of (-)-denopamine in atria and effects of low (-)-denopamine concentrations in papillary muscles and tracheae, as assessed with beta 1-selective antagonists. 2. High (-)-denopamine concentrations caused positive inotropic effects in papillary muscles and tracheorelaxant effects that were resistant to blockade by beta 1-, beta 2- and alpha-adrenoceptor antagonists (non-adrenergic effects). 3. (-)-Denopamine stimulated the adenylate cyclase of membranes derived from kitten ventricle and calf tracheal cells with an intrinsic activity of 0.3 and 0.2, respectively, compared to catecholamines. The contribution of beta 1- and beta 2-adrenoceptors to cyclase stimulation was assessed by selective blockade. Cyclase stimulation through beta 2-adrenoceptors by (-)-denopamine was 12% in ventricle and 82% in trachea but is not associated with positive inotropic effects and tracheal relaxation. 4. (-)-Denopamine exhibited only a 2- to 5-fold selectivity for beta 1-adrenoceptors compared to beta 2-adrenoceptors, as estimated consistently from binding assays and blockade of cyclase stimulation in myocardial and tracheal cell membranes. 5. Desensitization of kitten ventricular tissues, caused by a 3 h exposure to 30 mumol/l (-)-isoprenaline followed by 5 h washout, reduced the inotropic sensitivity of papillary muscles without decreasing the maximum inotropic effects of (-)-denopamine. In desensitized tissues, the nonadrenergic effect contributed by 30% to the maximum inotropic effect of (-)-denopamine. 6. In membranes, derived from desensitized tissues, the maximum adenylate cyclase stimulation induced by (-)-isoprenaline, (-)-denopamine and xamoterol was reduced to 1/2 of the corresponding stimulations observed in membranes from sham desensitized tissues. The density of beta-adrenoceptors, assessed with 3H-(-)-CGP 12,177, was not changed by the desensitization procedure suggesting that part of the receptors was uncoupled from the adenylate cyclase. The partial inotropic agonist xamoterol, which has an intrinsic activity of 0.5 in non-desensitized tissues, failed to cause positive inotropic effects in desensitized papillary muscles suggesting that not all cyclic AMP possesses inotropic relevance.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2566922 TI - Carbostyril-based beta-adrenergic agonists: evidence for long lasting or apparent irreversible receptor binding and activation of adenylate cyclase activity in vitro. AB - The interaction of the carbostyril derivatives 5-[2-[[1-(4-aminophenyl)-2-methyl prop-2-yl]amino]-1-hydroxyethyl]- 8-hydroxycarbostyril (carbo-amine) and 5-[2-[[3 [4-(bromoacetamido)phenyl]-2-methylprop-2-yl]amino]-1- hydroxyethyl]-8 hydroxycarbostyril (carbo-Br) with the rat reticulocyte beta-adrenoreceptor system has been partially characterized. In the absence of a guanine nucleotide, the concentration of carbo-amine, carbo-Br and (-)isoprenaline that inhibited (-) [125I]iodocyanopindolol ([125I]CYP) binding by 50% (IC50) was 5.9 +/- 0.2, 3.3 +/ 0.3 and 49 +/- 3 nM, respectively. In the presence of a guanine nucleotide, the IC50 values were carbo-amine, 21 +/- 0.6 nM; carbo-Br, 7.6 +/- 0.3 nM and ( )isoprenaline, 813 +/- 66 nM. Preincubation of membranes with either of the carbostyril congeners followed by washing reduced specific [125I]CYP binding capacity without changing the KD value for the remaining receptors. The beta antagonist nadolol largely prevented the receptor reduction induced by the carbostyril compounds. Incubation of membranes for 18 h at 25 degrees C resulted in an 11% recovery of the carbo-amine-induced receptor loss and no recovery of the receptors lost by preincubation with carbo-Br. However, the carbo-amine induced receptor loss could be largely reversed (80%) by membrane heating at 45 degrees C whereas little reversal (less than 10%) was observed with the carbo-Br pretreated membranes. The concentration of carbo-amine, carbo-Br and ( )isoprenaline that stimulated half-maximal cAMP formation in reticulocyte membranes was 17.8 +/- 3.1, 8.2 +/- 2.1 and 241 +/- 17 nM, respectively, and all 3 agonists produced the same maximal response. Initial cAMP formation stimulated by the carbostyril derivatives and (-)isoprenaline was blocked by concurrent addition of propranolol after 7 min of incubation with either of the two carbostyril derivatives did not affect further cAMP production whereas with ( )isoprenaline further cAMP production was blocked.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566923 TI - Involvement of central histaminergic and cholinergic systems in the morphine induced increase in blood-brain barrier permeability to sodium fluorescein in mice. AB - Morphine (5 mg/kg, s.c.) caused a submaximal increase in the brain level of sodium fluorescein administered i.v. Histamine H1-antagonists, diphenhydramine and mepyramine, given either i.p. or i.c.v., had no significant influence on the effect of morphine. H2-Antagonists, cimetidine and ranitidine, administered i.c.v., but not i.p., significantly inhibited the morphine effect. alpha Fluoromethylhistidine, a specific histidine decarboxylase inhibitor (given i.p. and i.c.v.) and antimuscarinic drugs, atropine and biperiden, but not methylatropine (given i.p.) also significantly reduced the morphine effect. Physostigimine (i.p.) significantly enhanced the effects of 0.5 and 1 mg/kg of morphine. Similar effects of histaminergic and cholinergic drugs were also observed on the buprenorphine- and DAGO-induced increase in blood-brain barrier (BBB) permeability to sodium fluorescein. None of the treatments with 6 hydroxydopamine, alpha-methyltyrosine, 5,7-dihydroxytryptamine or p chlorophenylamine had any significant effect on the morphine-induced increase in BBB permeability. These findings suggest that the activation of brain H2 receptors by neuronal histamine and muscarinic receptors by acetylcholine is involved in the increase in BBB permeability to sodium fluorescein caused by mu opioid receptor agonists. PMID- 2566924 TI - Differential increases in brain levels of neuropeptide Y and vasoactive intestinal polypeptide after kainic acid-induced seizures in the rat. AB - Changes in immunoreactivities of neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) were investigated in the brain of rats after severe kainic acid (KA, 10 mg/kg, i.p.) induced limbic seizures. Decreased levels of both neuropeptides were observed in the frontal cortex, straitum, dorsal hippocampus and amygdala/pyriform cortex subsequently to the period of acute seizures (3 h after injection of the toxin). Then NPY increased consistently in the frontal cortex, hippocampus and amygdala/pyriform cortex. Highest levels (290% of controls) were found in the frontal cortex after two months. Anticonvulsant therapy with phenobarbital (20 mg/kg, i.p., twice daily for three weeks) partially suppressed the rise in NPY levels. Immunoreactivity of VIP increased (to 150%) in the frontal cortex only transiently 3 days after injection of kainic acid. At the subsequently examined time intervals (10-60 days after kainic acid) it declined to control values. Levels decreasing subsequently to acute seizures reflect increased release and degradation of the respective peptide. Increased NPY levels suggest "upregulation" of NPY/somatostatin/GABA neurons due to the decreased seizure threshold of the animals. The early, reversible rise of VIP in the cortex points to a short-lasting activation of this peptide system contained in local cholinergic neurons. This may be a consequence either of the acute seizures or subsequent neuropathological changes. PMID- 2566925 TI - Down-regulation of 3H-lofentanil binding to opiate receptors in different cultured neuronal cells. AB - There was stereospecific binding of 3H-lofentanil (KD value = 1.53 nM) to membranes of neuroblastoma-glioma NG 108-15 cells which are known to bear high affinity binding sites for enkephalin derivatives (delta-opiate receptor subtype). There was no high affinity specific binding of the mu-opiate specific ligand 3H-sufentanil. The specific binding of 3H-lofentanil to delta-opiate receptor subtype was down-regulated (decrease in Bmax value without change in the KD value) after prolonged incubation of the cells in the presence of leu- and met enkephalin (0.1 microM). There was no down-regulation of the opiate receptors (3H-lofentanil and 3H-D-ala-D-leu-enkephalin specific binding) after incubation of NG 108-15 cells with drugs from the fentanyl series (alfentanil or sufentanil). In cultured neurones from rat forebrain (15 day old embryos), the 3H lofentanil binding was specific with high affinity (KD: 0.048 nM) and a slow dissociation rate similar to that in adult rat cortex. Drugs of the fentanyl series (4-anilino-piperidines) were potent displacers whereas agonists of the delta- (enkephalin derivatives), sigma- (phencyclidine, haloperidol, 3 hydroxyphenyl-propylpiperidine) or K- (U 50488) opiate sites had a low affinity (Ki greater than 0.5 microM) for 3H-lofentanil specific binding sites. Since there was also specific binding of 3H-sufentanil, the opiate receptors in cultured neurones seem to be mainly of the mu-subtype and this is consistent with the ontogeny of opiate receptors subtypes. These receptors were down-regulated after incubation in the presence of etorphine, sufentanil and alfentanil but not enkephalin derivatives.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566926 TI - Biphasic relaxant curves to glyceryl trinitrate in rat aortic rings. Evidence for two mechanisms of action. AB - The concentration-effect curve for the relaxant effects of glyceryl trinitrate (GTN) in rat aortic rings consisted of two phases with IC50 values of 0.1 microM for Phase I and 14 microM for Phase II. Incubation of tissues with oxyhaemoglobin or the induction of tolerance to GTN abolished responses occurring in Phase I but were without effect on Phase II relaxant responses. Both phases of the relaxant curve appeared to involve cyclic GMP since responses were (i) potentiated by the cyclic GMP phosphodiesterase inhibitor zaprinast (M & B 22948) and (ii) inhibited by methylene blue and LY83583, agents which inhibit soluble guanylate cyclase. The latter agents inhibited Phase I responses in a non-surmountable manner while Phase II responses were shifted to the right without effect on the maximal response. Neither phase of relaxation involved stimulation of the Na+/K+ ATPase pump since treatment of tissues with ouabain or K+-free solutions did not alter the GTN biphasic curve. Phase I relaxant responses to GTN resembled those to the endothelium-dependent relaxant acetylcholine, since oxyhaemoglobin and methylene blue were non-surmountable antagonists; however there was no cross tolerance to acetylcholine in GTN tolerant tissues. Phase II relaxant responses resembled those obtained with sodium nitroprusside (SNP) since neither oxyhaemoglobin nor the induction of tolerance to GTN altered the response to SNP. These results indicate that there are two distinct mechanisms of relaxation for GTN in rat aortic rings; however both mechanisms appear to involve cyclic GMP as the second messenger. PMID- 2566927 TI - Frequency-dependence of serotonin autoreceptor but not alpha 2-adrenoceptor inhibition of [3H]-serotonin release in rat hypothalamic slices. AB - The overflow of tritium from stimulated rat hypothalamic slices preincubated with [3H]-serotonin (5-HT) was significantly enhanced by reducing the frequency of stimulation from 3 Hz to 1 Hz while keeping the number of impulses constant. The 5-HT receptor agonist 5-methoxytryptamine inhibited in a concentration-dependent manner the electrically-evoked release of [3H]-5-HT with IC50 values of 560 nmol/l and of 34 nmol/l when the stimulations were delivered at 3 Hz and 1 Hz, respectively. The terminal 5-HT autoreceptor antagonist methiothepin enhanced in a concentration-dependent manner the electrically-evoked release of [3H]-5-HT and this effect was greater at a frequency of stimulation of 3 Hz than at 1 Hz. In the same paradigm, the 5-HT reuptake inhibitors citalopram and paroxetine did not alter the overflow of radioactivity elicited by stimulation at 3 Hz but significantly decreased it at 1 Hz. In the presence of 5-HT autoreceptor blockade achieved with methiothepin, citalopram increased the overflow of [3H]-5-HT to the same extent at 1 Hz and at 3 Hz. The IC50 values for inhibition of [3H]-5-HT release by the selective alpha 2-adrenoceptor agonist UK 14.304 were 35 nmol/l at 3 Hz and 30 nmol/l at 1 Hz. It is concluded that modulation of 5-HT release by 5 HT autoreceptors, but not by alpha 2-adrenoceptors is dependent on the synaptic concentration of 5-HT as a function of the frequency of depolarization. PMID- 2566928 TI - Characterization and presynaptic modulation of stimulation-evoked exocytotic co release of noradrenaline and neuropeptide Y in guinea pig heart. AB - The relationship between noradrenaline and neuropeptide Y (NPY) release was investigated in the in situ perfused guinea pig heart with intact sympathetic innervation. For determination of NPY concentrations in the perfusate, a specific radioimmunoassay was employed and further characterized. Electrical stimulation of the left stellate ganglion (4, 8, 12, and 50 Hz; for 10 min) evoked a calcium dependent and frequency-related overflow of noradrenaline and NPY, which was positively correlated (r = 0.83; p less than 0.001; n = 25). When two subsequent stimulations (12 Hz; each for 1 min) were performed in the same heart, addition of noradrenaline (10 microM) 5 min prior to the second stimulation reduced NPY overflow by 43 +/- 10%. The stimulated release of noradrenaline and NPY was increased by the alpha 2-adrenoceptor antagonist yohimbine (1 microM) to 170 +/- 10% and 199 +/- 26%, and attenuated by the alpha 2-adrenoceptor agonist B-HT 920 (1 microM) to 70 +/- 9% and 68 +/- 9%, respectively. The adenosine analogue cyclohexyladenosine (1 microM) significantly reduced the stimulated overflow of both noradrenaline (to 57 +/- 5%) and NPY (to 73 +/- 8%). Exogenous NPY (100 nM) attenuated the stimulated overflow of noradrenaline by 30 +/- 6%. Uptake1 blockade with desipramine (100 nM) or nisoxetine (100 nM) prior to the second stimulation significantly increased noradrenaline overflow and attenuated that of NPY; the attenuation of the stimulation-evoked overflow of NPY was abolished by yohimbine (1 microM). Our results indicate that electrical stimulation induces a calcium-dependent, exocytotic co-release of noradrenaline and NPY.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566930 TI - Presynaptic action of adrenaline on adrenoceptors modulating stimulation-evoked 3H-noradrenaline release from rabbit isolated aorta. AB - The purpose of this investigation was to study the effect of adrenaline on presynaptic adrenoceptors by recording the release of 3H-noradrenaline evoked by electrical-field stimulation. Adrenaline (10(-10)-3 x 10(-9) mol/l) had no effect on the 3H-overflow evoked by stimulation of aorta preloaded with 3H noradrenaline. At 10(-8) and 3 x 10(-8) mol/l, the 3H-overflow was decreased by up to 47%. The maximum decrease was more marked in the presence of either cocaine (3 x 10(-5) mol/l) plus corticosterone (4 x 10(-5) mol/l), cocaine (3.3 x 10(-6) mol/l) plus normetanephrine (4 x 10(-5) mol/l), or desipramine (10(-6) mol/l) plus normetanephrine (10(-5) mol/l). The relationship between adrenaline-induced decrease and stimulation-frequency was dependent on the experimental design: either the decrease was the same at all frequencies (1-16 Hz) or it was more marked, the lower the frequency (1 greater than 3 greater than 8 Hz). Phentolamine and rauwolscine (both 10(-6) mol/l) antagonized the inhibitory effect of adrenaline (10(-8)-10(-6) mol/l). Phenoxybenzamine (10(-6) mol/l), prevented the inhibitory effect. No enhancing effect of adrenaline (10(-9)-10(-6) mol/l) was observed in the presence of these three alpha-adrenoceptor antagonists. Our results suggest that adrenaline activates inhibitory alpha 2 adrenoceptors, but not facilitatory beta-adrenoceptors on postganglionic sympathetic nerve terminals in rabbit aorta. PMID- 2566929 TI - Neuronally released (-)-noradrenaline relaxes smooth muscle of calf trachea mainly through beta 1-adrenoceptors: comparison with (-)-adrenaline and relation to adenylate cyclase stimulation. AB - The nature of the receptors that mediate the relaxation of smooth muscle by field stimulation, (-)-noradrenaline and (-)-adrenaline was investigated in calf tracheal smooth muscle. The relation between relaxation, stimulation of the adenylate cyclase and density of beta-adrenoceptor subtypes was studied with the help of antagonists of beta 1- and beta 2-adrenoceptors. The question of the existence of catecholamine-containing nerves was also investigated. (1) Nerves with varicosities exhibiting catecholaminergic fluorescence were observed between bundles of smooth muscle cells. (2) Consistent with the existence of adrenergic nerves (-)-noradrenaline was also found. The content of (-)-noradrenaline (1 microgram.g-1 w.w.) was the same in smooth muscle strips from the sublaryngeal region and from the region close to the bifurcation of the calf trachea. (-) Adrenaline was not detected. (3) Smooth muscle relaxation by low (-) noradrenaline concentration (0.6-2 nmol/l) was mediated through beta 1 adrenoceptors. Low concentrations of (-)-adrenaline (0.06-1 nmol/l) relaxed through beta 2-adrenoceptors. High concentrations of (-)-noradrenaline and (-) adrenaline also caused relaxation through beta 2- and beta 1-adrenoceptors respectively. (4) Field stimulation caused relaxation which was half maximal at 0.2-0.8 Hz. Blockade of beta 1-adrenoceptors strongly attenuated the relaxant response to field stimulation and shifted the frequency-relaxation curves to 4 times higher frequencies. These results are consistent with a beta 1-adrenoceptor mediated relaxation caused by (-)-noradrenaline released from sympathetic nerve endings at low stimulation frequencies. (5) Blockade of beta 2-adrenoceptors failed to reduce smooth muscle relaxation caused by field stimulation at low stimulation frequencies (0.1-1 Hz). However, after beta 1-adrenoceptor blockade, additional blockade of beta 2-adrenoceptors reduced the relaxant effects observed at high frequencies (2-400 Hz). The results suggest that high concentrations of endogenous (-)-noradrenaline cause relaxation through beta 2-adrenoceptors. (6) Binding experiments with 3H-(-)-bupranolol and 3H-ICI 118,551 revealed between 10,000 and 20,000 beta-adrenoceptors per smooth muscle cell of which 3/4 were beta 2 and 1/4 beta 1. The equilibrium dissociation constant of (-)-adrenaline for both beta 1- and beta 2-adrenoceptors and of (-)-noradrenaline for beta 1 adrenoceptors was 1 mumol/l. The affinity of (-)-noradrenaline for beta 2 adrenoceptors was 10 to 20 times lower than for beta 1-adrenoceptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566931 TI - Uptake inhibitors do not change the effect of imidazoline alpha 2-adrenoceptor agonists on transmitter release evoked by single pulse stimulation in mouse vas deferens. AB - The present study was undertaken to compare the presynaptic interaction of neuronal noradrenaline uptake inhibitors with imidazoline and phenylethylamine alpha 2-adrenoceptor agonists under two different conditions: at low and high noradrenaline concentrations in the biophase. Isolated mouse vasa deferentia were stimulated with trains of 7 pulses given at 0.2 Hz and the inhibition by the alpha 2-adrenoceptor agonists clonidine, alpha-methylnoradrenaline, and UK-14,304 of twitch responses was measured in the absence and in the presence of either cocaine (12 mumol/l) or desipramine (40 nmol/l). The effects were determined for the first (equivalent to single pulse stimulation) and the last stimulus of each train. Both uptake inhibitors antagonized the presynaptic inhibitory effects of imidazolines (clonidine and UK-14,304) on the last twitch; the effects on the first twitch remained unchanged. In contrast, the uptake inhibitors potentiated the inhibitory effect of the phenylethylamine (alpha-methylnoradrenaline) on both the first and the last twitches. These results support the view that the concentration of noradrenaline in the biophase plays a decisive role in the inhibition by alpha 2-adrenoceptor agonists of the electrically evoked release of noradrenaline. Agonists that are not substrates of neuronal uptake (i.e., clonidine, UK-14,304) become less effective when noradrenaline is present in the biophase while substrates of neuronal uptake (i.e., alpha-methylnoradrenaline) do not. The results argue against the hypothesis that uptake inhibitors interact directly with presynaptic alpha 2-adrenoceptors or act at some link between uptake and receptor sites. PMID- 2566932 TI - Excitatory amino acid receptors in rat locus coeruleus. An extracellular in vitro study. AB - The goal of this study was to investigate whether locus coeruleus neurons of the rat are sensitive to agonists of the different excitatory amino acid receptors. All experiments were performed on a midpontine rat slice preparation. Bath applied L-glutamate, kainate, N-methyl-D-aspartate (NMDA) and quisqualate induced concentration-dependent activations of all neurons which were reflected in an increase of the neurons' mean discharge rate. The rank order of cell activation was kainate approximately quisqualate greater than NMDA greater than L-glutamate. None of the agonists induced a bursting-type of discharge. The NMDA-receptor blocker DL-2-amino-5-phosphonovaleric acid (APV, 30 microM) selectively antagonized the NMDA-induced increase in cell firing. Kynurenic acid (100 microM) non-selectively attenuated the response to NMDA, kainate and quisqualate. Neither APV nor kynurenic acid per se had any effect on the spontaneous firing rate. If the Mg2+ concentration in the superfusion medium was lowered from 2 mM to nominally zero the response to NMDA was selectively increased. In conclusion, locus coeruleus neurons share with other neurons their sensitivity to agonists of all three types of excitatory amino acid receptors. However, in contrast to other neurons, they do not respond with a bursting type of discharge. PMID- 2566933 TI - Effect of omega-conotoxin GVIA on release of 3H-gamma-aminobutyric acid from slices of rat neostriatum. AB - omega-Conotoxin GVIA (omega-CT) diminished the potassium-induced in vitro release of 3H-gamma-aminobutyric acid (3H-GABA) from slices of rat neostriatum in a manner which depended on the concentration of potassium. omega-CT (0.1 mmol/l) decreased the release of 3H-GABA induced by 25 mmol/l K+ from 11.6% to 6.1% of tissue content, ie. by 48%, while it did not affect the release of 3H-GABA caused by 20 mmol/l K+, which was 4.8% of tissue content. However, in the presence of a polyclonal antiserum or cysteamine (600 mumol/l), both of which diminish the effects of endogenous somatostatin, 0.1-10 nmol/l omega-CT decreased the release of 3H-GABA induced by 20 mmoles/l K+ by 40%. It is concluded that omega-CT did not only inhibit GABA-neurones, but had an additional inhibitory effect on somatostatin neurones which are known to depress the release of 3H-GABA. It is further concluded that neuronal interactions, which are possible in brain slice preparations, may impede the interpretation of effects of drugs, especially if agents are used which affect basic mechanisms of transmitter release and thus the release of various transmitters from neurones. PMID- 2566935 TI - Peptic blockade. PMID- 2566936 TI - [Psychopathology and cognitive disorders in chronic schizophrenic patients with tardive dyskinesia]. PMID- 2566934 TI - Preponderance of beta- over alpha-adrenoceptors in mediating the positive inotropic effect of phenylephrine in the ferret ventricular myocardium. AB - [3H]prazosin bound to the membrane fraction derived from the ferret ventricular muscle with high affinity in a saturable manner (Kd = 0.25 nmol/l and Bmax = 27 fmol/mg protein in the right ventricle). [3H]CGP-12177, a beta-adrenoceptor ligand, bound to the membrane fraction with a Kd value of 0.29 nmol/l and a Bmax of 42 fmol/mg protein. In the isolated ferret papillary muscle driven at 1 Hz at 37 degrees C, phenylephrine elicited a concentration-dependent positive inotropic effect. The maximal effect of phenylephrine was comparable to that of isoprenaline. Prazosin (0.3 mumol/l) shifted the concentration-response curve for phenylephrine slightly but significantly to the right, the maximal response being unaffected. In contrast, bupranolol (0.3 mumol/l) shifted the curve for phenylephrine markedly downwards: the maximal response was depressed significantly to 40% and the curve became less steep. In the presence of prazosin and bupranolol the curve was shifted to the right, being essentially parallel to the control curve. These results indicate that in the ferret ventricular myocardium both alpha- and beta-adrenoceptors mediate the positive inotropic effect of phenylephrine. The extent of contribution of the two classes of adrenoceptor is quite different from that in other mammalian species. In the ferret heart, beta-adrenoceptors predominate over alpha-adrenoceptors in mediating the positive inotropic effect of phenylephrine, although the number of beta-adrenoceptors is not especially high when compared with other species. PMID- 2566937 TI - Takayasu's arteritis and nephrotic syndrome in a patient with crossed renal ectopia. AB - A male patient with nephrotic syndrome, crossed renal ectopia, and extensive Takayasu's arteritis (TA), involving one of the renal arteries, is described. Renal histology showed amyloidosis. After reviewing the literature and considering the finding of raised serum amyloid protein A levels, it was concluded that TA had probably caused the amyloidosis. The crossed renal ectopia must have been a purely coincidental finding. PMID- 2566938 TI - First International Conference on Alzheimer's Disease and Related Disorders. PMID- 2566939 TI - Calmodulin selectively modulates the guanylate cyclase activity by repressing the lipid phase separation temperature in the inner half of the bilayer of rat brain synaptosomal plasma membranes. AB - The association of [125I-]calmodulin with rat brain synaptosomal plasma membranes, when incubated for 1 h at 25 degrees in the presence or in absence of 20 microM Ca2+, follows a sigmoid path with a Hill coefficient h = 1.79 +/- 0.12 and h = 1.72 +/- 0.11, respectively. The total association of calmodulin with the membrane increased approx. 60%-80% at all the range of calmodulin concentrations used in the presence of 20 microM Ca2+. A three fold increase of guanylate cyclase activity was shown in the presence of low concentrations of calmodulin (up to 10 nM); higher concentrations (up to 40 nM) however, led to a progressive inhibition of the enzyme activity with respect to maximal stimulation. Calmodulin increased the lipid fluidity of synaptosomal plasma membranes labeled with 1,6 diphenyl-1,3,5-hexatriene (DPH), as indicated by the steady-state fluorescence anisotropy [(ro/r)-1]-1. Arrhenius-type plots of [(ro/r)-1]-1 indicated that the lipid separation of the membrane at 22.7 +/- 1.2 degrees was perturbed by calmodulin such that the temperature was reduced to 16.3 +/- 0.9 degrees and 15.5 +/- 0.8 degrees in the absence or in the presence of 20 microM Ca2+. Arrhenius plots of guanylate cyclase and acetylcholinesterase activities exhibited break points at 26.7 +/- 1.4 degrees and 22.3 +/- 1.0 degrees in control synaptosomal plasma membranes, respectively. The break point for the guanylate cyclase was reduced to 16.3 +/- 0.9 degrees in calmodulin treated synaptosomal plasma membranes whereas that of acetylcholinesterase remained unaffected (21.1 +/- 0.9 degrees).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566941 TI - Dopaminergic agents differentially regulate both processing and content of alpha N-acetylated endorphin and alpha-MSH in the ovine pituitary intermediate lobe. AB - Hypothalamo-pituitary disconnection (HPD) in the sheep results in a two-fold increase in pituitary intermediate lobe (IL) immunoreactive (ir)-alpha-N acetylated endorphin (NacEP) and ir-alpha-melanocyte-stimulating hormone (alpha MSH) content. The rise in IL NacEP content is accompanied by a markedly altered pattern of processing, in that NacEP1-27 becomes the dominant molecular species with a complementary fall in Nac alpha-EP and Nac gamma-EP. To determine if these effects reflect the loss of descending dopaminergic neuronal input to the IL, we have chronically treated two groups (n = 4 per group) of normal sheep with the dopamine antagonist haloperidol or the dopamine agonist bromocriptine; a group of HPD sheep were also treated with bromocriptine. Acid extracts of IL were diluted and ir-alpha-MSH and ir-NacEP content determined by radioimmunoassay; aliquots were submitted to reversed-phase HPLC and collected fractions similarly assayed. Bromocriptine lowered ir-NacEP and ir-alpha-MSH by about 30%; on HPLC the ir NacEP profiles, and perhaps to a lesser extent those for ir-alpha-MSH were qualitatively similar to untreated controls. In contrast, haloperidol increased by about 45% both ir-NacEP and ir-alpha-MSH levels and produced a marked change in the ir-NacEP molecular profile, with NacEP1-27 becoming the predominant molecular form and other species representing only minor components in the chromatogram. In the treated HPD group, bromocriptine partially restored the processing profiles previously observed in HPD animals to those found in untreated intact animals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566940 TI - The effects of transmitters on the affinity of the insoluble fraction of ox brain for Na+ and K+. AB - The insoluble fraction of ox-brain, which had previously been shown to have a non linear affinity for Na+ and K+, was prepared. Acetylcholine (1 x 10(-8) mol/l and 1 x 10(-7) mol/l) reduced the affinity of the fraction for Na+ and K+ to zero, while at 1 x 10(-6) mol/l, the affinity for the cations was almost as high as in the absence of the transmitter; the affinities for Na+ and K+ were particularly high, when the supernatant concentrations of these ions exceeded 80-100 mM. Addition of eserine (3 x 10(-5) mol/l) considerably modified the response of the fraction to acetylcholine (1 x 10(-5) mol/l). Atropine (1 x 10(-8) mol/l) in the absence or presence of acetylcholine (1 x 10(-5), or 1 x 10(-4) mol/l) reduced the affinity of the fraction for Na+ and K+ to zero. Epinephrine (3 x 10(-10) mol/l) lowered the affinity for Na+ and K+, while ergotamine itself (1 x 10(-5) mol/l) reduced it to zero. The addition of both epinephrine and ergotamine at the latter concentrations restored the affinity of the fractions for Na+ and K+ to what it had been in the absence of the transmitter or antagonist, previously reported. Norepinephrine (3 x 10(-10) mol/l), or ouabain (1 x 10(-7) mol/l) reduced the affinity of the fraction for Na+ and K+ to zero. Thus, the transmitters and antagonists altered the affinity of the insoluble fraction for Na+ and K+ non-linearity, dependent upon their concentrations, the concentrations of the cations, and the interaction of transmitter and antagonist. PMID- 2566942 TI - Activation of cholinergic neurotransmission by pyridostigmine reverses the inhibitory effect of hyperglycemia on growth hormone (GH) releasing hormone induced GH secretion in man: does acute hyperglycemia act through hypothalamic release of somatostatin? AB - Acute hyperglycemia blocks growth hormone (GH) secretion in response to provocative stimuli including growth hormone releasing hormone (GHRH) administration. However, the precise mechanism of glucose action is unknown. To determine if enhanced somatostatinergic stimulation accounts for the decreased GH secretion, we studied the effect of enhanced cholinergic tone by pyridostigmine on the hyperglycemia blockade of GH release in 7 normal subjects. Pyridostigmine, an acetylcholinesterase inhibitor, has been postulated as an inhibitor of somatostatin release. Each subject underwent 4 tests with GHRH injection (100 micrograms i.v. at 0 min). In the first (control) test, placebo was administered before GHRH. In the second test, 100 g of glucose was administered p.o. 45 min before GHRH. In the third test, pyridostigmine, 120 mg p.o., was administered 60 min before GHRH, and in the fourth test, pyridostigmine, glucose and GHRH were administered at -60, -45 and 0 min, respectively. GHRH-induced GH secretion of 25.8 +/- 4.5 ng/ml was significantly reduced by previous glucose administration (12.1 +/- 4.5 ng/ml) and significantly potentiated by previous pyridostigmine pretreatment (56.5 +/- 16.8 ng/ml). In the fourth test (pyridostigmine plus glucose plus GHRH) the GH peak of 42.4 +/- 9.2 ng/ml was significantly higher than after GHRH alone and not different to the pyridostigmine-GHRH test. In conclusion, central cholinergic activation by pyridostigmine reversed the hyperglycemic blockade of GHRH-induced GH secretion. In addition, hyperglycemia was unable to reduce the potentiating effect of pyridostigmine on GH secretion elicited by GHRH. Based on the reported actions of pyridostigmine, acute hyperglycemia might act over GH release by inducing hypothalamic somatostatin release. PMID- 2566943 TI - Calcium dependence of somatostatin (SRIF) release and cyclic AMP levels in cultured diencephalic neurons. AB - Calmodulin has been reported to be involved in the Ca2+-dependent hypothalamus in vitro. The present experiments were undertaken to determine whether at an early stage of development (in diencephalic primary cultures secreting SRIF, on the 11th day) the activation of a Ca2+-calmodulin kinase system is also involved in the release of the peptide. Since a calmodulin-dependent adenylate cyclase activity has been detected in the brain, we measured intracellular cyclic AMP accumulation as an additional parameter of calmodulin activity. SRIF release and cyclic AMP accumulation were stimulated by K+ (56 mM) and by the Ca2+ ionophores ionomycin (0.5 microM) and A 23187 (in a dose-dependent manner). Incubation of cells in Ca2+-free Locke medium or in the presence of Co2+ (1 mM) completely blocked ionophore-induced SRIF release and cyclic AMP accumulation. Three calmodulin antagonists (calmidazolium, W-7, and chlorpromazine) and two blockers of calmodulin-dependent kinase (phenytoin and diazepam) were tested on evoked SRIF release and cyclic AMP formation. Neither W-7 nor calmidazolium modified A 23187-induced SRIF release at any dose tested, although they inhibited, in a dose dependent manner, the stimulatory effect of the Ca2+ ionophore on cyclic AMP accumulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566944 TI - Antagonists of adenosine and alpha-2-adrenoceptors reverse the anticonvulsant effects of tizanidine in DBA/2 mice. AB - The anticonvulsant activity of 5-chloro-4-(2-imidazolin-2yl-amino)-2,1,3 benzothiazole, tizanidine, was studied following intraperitoneal (i.p.) administration in DBA/2 mice (which show sound-induced seizures). Protection against sound-induced seizures was observed after tizanidine, (0.5-4 mg/kg i.p.). The ED50 values for suppression of the tonic, clonic and wild running phases of sound-induced seizures were 0.54, 0.76 and 1.43 mg/kg, respectively. This protective action was significantly reduced by pretreatment with aminophylline (25 mg/kg i.p.), yohimbine (1 mg/kg i.p.) or piperoxan (20 mg/kg i.p.). Methysergide, a serotonin antagonists, did not significantly reduce the anticonvulsant effects of tizanidine. The present experiments suggest an involvement of purinergic and adrenergic mechanisms in the anticonvulsant action of tizanidine. PMID- 2566945 TI - Interactions of alpha 2-adrenoceptor antagonists with medetomidine and with ethanol in a holeboard test. AB - The effects of two selective alpha 2-adrenoceptor agonists (clonidine and medetomidine) and antagonists (atipamezole and idazoxan) were examined in the holeboard test. The interactions of the two antagonists with ethanol were also investigated. Atipamezole (0.1-3.0 mg/kg) and idazoxan (0.01-0.3 mg/kg) were without effect on either directed exploration or locomotor activity in the holeboard test, whereas clonidine (0.003-0.1 mg/kg) and medetomedine (0.003-0.1 mg/kg) were sedative. Atipamezole (1-3 mg/kg) and idazoxan (0.3-1.0 mg/kg) reversed the behavioral effects of 0.1 mg/kg of medetomidine. When administered with ethanol (2 g/kg), atipamezole (1-3 mg/kg) showed a significant antagonism of the ethanol-induced reduction in exploratory head-dipping: no change in the locomotor stimulant properties of ethanol was seen. A similar trend was seen for exploration after a combination of idazoxan (1-3 mg/kg) with 2 g/kg ethanol; however, the 3 mg/kg dose attenuated the locomotor stimulant effect. Both antagonists caused a dose-related reduction in the increase in head-dipping seen after administration of 1 g/kg of ethanol, without any effect on the locomotor stimulant effect of this dose. These results suggest mediation of alpha 2 adrenoceptors in some of the behavioral effects of ethanol. PMID- 2566946 TI - Conditioned taste aversions induced by phencyclidine and other antagonists of N methyl-D-aspartate. AB - Taste aversions can be conditioned in rats by a variety of psychoactive drugs, including those with reinforcing properties. Previous research, however, has not established clearly whether phencyclidine and related drugs are active in such procedures. The present study was carried out to investigate whether phencyclidine would induce a conditioned taste aversion and whether several other compounds (MK-801, the stereoisomers of NANM and ifenprodil) which, like phencyclidine, are known to antagonise the actions of N-methyl-D-aspartate (NMDA), would produce similar effects. When rats received injections of these compounds, after consuming a novel solution of saccharin, their subsequent consumption of the same solution decreased. The smallest doses of the different drugs which induced clear taste aversions were: phencyclidine 3 mg/kg, MK-801 0.3 mg/kg, (+)-NANM 10 mg/kg, (-)-NANM 3 mg/kg and ifenprodil 10 mg/kg. Thus, all the drugs were active. However, as neither the potencies nor the efficacies of the different compounds in inducing taste aversions correlated with their other behavioural effects or with their relative potencies in antagonising the effects of NMDA or in displacing phencyclidine from its binding sites, the mechanisms involved are unclear. PMID- 2566947 TI - Evidence that central 5-HT2 receptors do not play an important role in the anorectic activity of D-fenfluramine in the rat. AB - To gain information on the role of central 5-HT2 receptors in the reduction of food intake caused by D-fenfluramine in rats, different intraperitoneal doses of metergoline, a non-selective 5-HT receptor antagonist and ritanserin, a selective 5-HT2 receptor antagonist, were compared for their ability (a) to antagonize the anorectic effect of D-fenfluramine; (b) to occupy central 5-HT2 receptors in vivo (measured by the binding of [3H]spiperone in the frontal cortex) and (c) to affect the concentrations of D-fenfluramine and its active metabolite, D norfenfluramine in brain. Metergoline dose-dependently reduced the effect of D fenfluramine (2.5 mg/kg i.p.) on food intake, with complete antagonism at 1 mg/kg, a dose which occupies about 50% of cortical 5-HT2 receptors. Ritanserin, at a dose (0.5 mg/kg) causing 50% occupation of 5-HT2 receptors, had no effect on anorexia induced by D-fenfluramine and only partially prevented it at doses which caused maximum occupation of 5-HT2 receptors (1-2 mg/kg). Unlike 1 mg/kg metergoline, 1 mg/kg ritanserin significantly reduced the concentrations of D norfenfluramine in the frontal cortex and hypothalamus of rats 30 min after injection of D-fenfluramine. The results suggest that 5-HT receptors, other than 5-HT2, possibly 5-HT1B, are involved in the anorectic effect of D-fenfluramine in food-deprived rats. PMID- 2566948 TI - Effect of 5-HT1A agonists on stress-induced deficit in open field locomotor activity of rats: evidence that this model identifies anxiolytic-like activity. AB - Deficits in locomotion and exploratory behaviour in an open field were induced in rats by restraint for 2 hr, 23 hr before testing. Diazepam, 0.62 and 1.25 mg/kg, intraperitoneally (i.p.), 15 min before testing, reversed the stress-induced reduction in locomotion; 1.25 mg/kg also attenuated the effect of stress on exploration (rearing and object exploring). Diazepam did not affect the activity of controls. A putative anxiogenic compound, pyrazoloquinoline (CGS 8216, 10 mg/kg administered 30 min before testing), also markedly reduced locomotion and exploration and the effect was reversed by 2.5 mg/kg diazepam, 15 min before testing. Buspirone, 0.1 mg/kg subcutaneously (s.c.) 15 min before testing, significantly attenuated the effect of stress on locomotion and exploration but had no effect in controls. Larger doses (0.5 and 1.0 mg/kg) markedly reduced the behavioural measures in controls and did not modify or enhance the effect of stress. 8-Hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), 0.25 and 0.5 mg/kg (s.c.), 1 hr before testing, significantly attenuated the reduction in locomotion without affecting rearing and object-exploring in stressed rats. At doses from 0.125 to 0.5 mg/kg, 8-OH-DPAT reduced exploration in control rats. Two hr after restraint (corresponding to 21 hr before testing in the open field) 8-OH-DPAT, 0.125 to 2 mg/kg (s.c.), did not modify the open field deficits, caused by stress. In these treatment conditions, 0.5 and 2 mg/kg 8-OH-DPAT reduced locomotion and exploration in control rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566950 TI - The effects of anxiolytic and anxiogenic benzodiazepine receptor ligands on motor activity and levels of ascorbic acid in the nucleus accumbens and striatum of the rat. AB - The effects of the anxiolytic benzodiazepine flurazepam and the anxiogenic beta carboline N-methyl-beta-carboline-3-carboxylate (FG 7142) were measured in unanaesthetised rats. Changes in motor activity, using a Doppler-shift microwave device, and in the extracellular concentration of ascorbate in the striatum and nucleus accumbens, using linear sweep voltammetry with carbon paste electrodes, were monitored continuously over a period of 7 days. Both motor activity and release of ascorbate were greater during the dark than the light period; regression analysis showed a high correlation coefficient for motor activity vs release of ascorbate. The two drugs caused similar changes in this diurnal pattern. A single intraperitoneal injection of either flurazepam or FG 7142 at the end of the light period was followed by a reduction in the nocturnal rise of motor activity and of levels of ascorbate in both the nucleus accumbens and striatum. However, whereas the correlation coefficient for motor activity vs the level of ascorbate in both the nucleus accumbens and striatum remained high after the injection of flurazepam, there was a breakdown of the correlation on the day after the injection of FG 7142, followed by recovery. PMID- 2566949 TI - Fentanyl produces cholinergically-mediated analeptic and EEG arousal effects in rats. AB - Fentanyl (20 micrograms/kg i.p.), administered to naltrexone-pretreated, pentobarbital-anesthetized rats, produced a shortening of the duration of narcosis. This analeptic effect was blocked by atropine, but not by methylatropine, indicating that a central cholinergic mechanism was involved. Fentanyl also increased sodium-dependent high affinity uptake of choline activity in the hippocampus and cortex that had been depressed by the barbiturate. Injection of 0.8 ng of fentanyl into the pontis oralis in the pontine reticular formation also produced analepsis in naltrexone-pretreated, pentobarbitalized rats. Hippocampal EEG recordings also showed the appearance of cholinergically mediated theta activity, which was indicative of arousal activity in the hippocampus. These results suggest that fentanyl, in addition to possessing potent opiate activity, also activates a nonopioid-mediated central cholinergic arousal system. PMID- 2566951 TI - Central regulation of adrenal tyrosine hydroxylase: interaction between dopamine and GABA systems. AB - It has been previously demonstrated that nigrostriatal dopaminergic fibres participate in the neural regulation of the activity of adrenal tyrosine hydroxylase, specifically in its induction. To determine whether activation or inhibition of these fibres is responsible for this induction, the role of presynaptic dopamine receptors was investigated. Apomorphine (0.2 mg/kg), (+)3 PPP (10 mg/kg) and BHT 920 (1-3 mg/kg), drugs that are reported to bind to presynaptic dopamine receptors and thereby inhibit the release of that neurotransmitter, caused significant increases in the activity of the enzyme. As a central GABA (gamma-aminobutyric acid) system is believed to exert inhibitory control over the release of dopamine, GABA agonists were also tested for their effects. Muscimol (3 mg/kg), gamma-hydroxybutyrate (500 mg/kg) and HA-966 (150 mg/kg) produced significant induction of the adrenal enzyme; this induction was not blocked by dopamine postsynaptic receptor antagonists. After intraventricular administration (5 micrograms/rat) in normal animals, HA-966 produced significant induction of tyrosine hydroxylase. Its systemic administration did not induce the enzyme in animals with the adrenal denervated. When administered together at submaximal doses, HA-966 and BHT 920 produced an additive effect in the induction of adrenal tyrosine hydroxylase. PMID- 2566952 TI - Neuropeptides modulate beta-adrenergic agonist binding and receptor downregulation in slices of rat brain. AB - Displacement curves of the beta-adrenergic agonist (-)-isoproterenol, on the binding of [3H]-dihydroalprenolol (DHA) in homogenates from the brain of rat defined a two-site model of high (KH; 8.5 +/- 1.6 nM) and low (KL; 771 +/- 111 nM) affinity. The KL/KH ratio was 84.5 +/- 4.5. Somatostatin, ACTH, dynorphin and mastoparan, but not vasoactive intestinal peptide (VIP), substance P, met enkephalin or leu-enkephalin, each at 1 microM, decreased the KL/KH ratio to 35.2 +/- 4.2, 43.0 +/- 5.0, 52.0 +/- 2.3 and 60.3 +/- 8.0, respectively, without significant effect on either the Kd or Bmax of antagonist binding. Calcium was required for this effect of the neuropeptides. In slices from whole brain of rat, incubated for 2 hr, ACTH attenuated the isoproterenol-induced downregulation of beta-adrenoceptors. For beta-adrenergic agonists, there appeared to be a close correlation between the KL/KH ratio and intrinsic activity. These results suggest that certain neuropeptides may influence beta-adrenergic neurotransmission by modulating the intrinsic activity of beta-agonists. This modulation is manifested, at least in part, by changes in both binding affinity and receptor down-regulation. PMID- 2566953 TI - Neurochemical characteristics of aluminum-induced neurofibrillary degeneration in rabbits. AB - Aluminum-induced neurofibrillary degeneration in rabbits is known to affect particular populations of neurons. The neurotransmitter alterations which accompany aluminum neurofibrillary degeneration were examined in order to assess how closely they mimic those of Alzheimer's disease. There was a significant reduction in choline acetyltransferase activity in entorhinal cortex and hippocampus as well as significant reductions in cortical concentrations of serotonin and norepinephrine in the aluminum-treated rabbits. Significant reductions in glutamate, aspartate and taurine were found in frontoparietal and posterior parietal cortex. Concentrations of GABA were unchanged in cerebral cortex. Both substance P and cholecystokinin immunoreactivity were significantly reduced in entorhinal cortex but there were no significant changes in somatostatin, neuropeptide Y and vasoactive intestinal polypeptide. The five neuropeptides were unaffected in striatum, thalamus, cerebellum and brainstem. Neurochemical changes were found in the regions with the most neurofibrillary degeneration while regions with little or no neurofibrillary degeneration were unaffected. The reductions in choline acetyltransferase activity, serotinin and noradrenaline suggest that some neuronal populations preferentially affected in Alzheimer's disease are also affected by aluminum-induced neurofibrillary degeneration; however, the cortical somatostatin deficit which is a feature of Alzheimer's disease is not replicated in the aluminum model. PMID- 2566954 TI - The colocalization of cholecystokinin and tyrosine hydroxylase mRNAs in mesencephalic dopaminergic neurons in the rat brain examined by in situ hybridization. AB - The colocalization of cholecystokinin and tyrosine hydroxylase mRNAs was studied with a cellular resolution in the mesencephalic dopaminergic neurons of the rat brain by in situ hybridization using synthetic oligonucleotides. An extensive colocalization of cholecystokinin-expressing cells, greater than that seen previously by immunohistochemistry, was found in the ventral tegmental area and in the substantia nigra pars compacta. We observed in these regions that cholecystokinin and tyrosine hydroxylase mRNAs coexisted in the same neurons but not all dopamine cells expressed cholecystokinin mRNA. 6-Hydroxydopamine-induced destruction of mesostriatal dopaminergic neurons resulted in a complete loss of cholecystokinin and tyrosine hydroxylase mRNA expression throughout the substantia nigra pars compacta, indicating that all cholecystokinin expressing cells are 6-hydroxydopamine-sensitive. While increased enkephalin mRNA expression in the striatum ipsilateral to the lesion was detected, no change of cholecystokinin mRNA expression was observed in any forebrain on the lesioned side, suggesting that cholecystokinin expression in the forebrain is not under dopaminergic control. These results show the usefulness of the in situ hybridization approach for the precise localization of cells in rat brain which express mRNAs for cholecystokinin and tyrosine hydroxylase and for the study of the effects of neurotoxic lesions on these cells. PMID- 2566955 TI - Synaptic activation of N-methyl-D-aspartate and non-N-methyl-D-aspartate receptors in the mossy fibre pathway in adult and immature rat cerebellar slices. AB - The participation of excitatory amino acid receptors in mossy fibre-granule cell synapses in lobule VIa of adult and immature rat cerebellar slices was investigated using an extracellular grease-gap technique. For the immature slices, the age selected (14 days after birth) was one at which the sensitivity of granule cells to exogenous N-methyl-D-aspartate is much higher than in the adult. The principal synaptic potentials observed after low-frequency electrical stimulation of the white matter resembled closely those found to be centred in the granule cell layer in field potential studies in the cat in vivo. They comprised a short latency negative potential, a slow negative wave and, in the adult, a further late negative wave. In the adult, with 1.2 mM Mg2+ in the perfusing solution, none of these potentials was significantly affected by the N methyl-D-aspartate antagonist, 2-amino-5-phosphonovalerate, but they were all markedly inhibited by the broad spectrum antagonist, kynurenate, and, more potently, by the selective non-N-methyl-D-aspartate receptor blocker, 6-cyano-2,3 dihydroxy-7-nitro-quinoxaline. After removal of Mg2+, which has a blocking action on the ion channels associated with N-methyl-D-aspartate receptors, the size of all the potentials increased. The increase in the short latency potential was insensitive to 2-amino-5-phosphonovalerate but a component of the slow negative wave (and of the late negative wave) was reduced back to control levels by the antagonist. Application of 6-cyano-2,3-dihydroxy-7-nitro-quinoxaline (10 microM) in Mg2+-free solution revealed, in near isolation, a slow wave (latency to peak, 28 ms) which could be abolished by 2-amino-5-phosphonovalerate. In the immature slices, bathed in normal (Mg2+-containing) medium, 2-amino-5-phosphonovalerate caused a small reduction in the short latency potential and inhibited a component of the slow negative wave which could, again, be observed in relative isolation after perfusion of 6-cyano-2,3-dihydroxy-7-nitro-quinoxaline. Removal of Mg2+ increased the amplitudes of the short latency potential and the slow negative wave in a manner which was sensitive to 2-amino-5-phosphonovalerate and increased the size of the slow, 6-cyano-2,3-dihydroxy-7-nitro-quinoxaline-resistant wave. It is concluded that glutamate is likely to be the transmitter released by mossy fibres, at least those innervating lobule VIa.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2566956 TI - Release of taurine from cultured cerebellar granule cells and astrocytes: co release with glutamate. AB - The properties of the release of preloaded [3H]taurine and endogenous taurine were studied with cultured cerebellar granule cells (7-8 days in vitro) and astrocytes (14-15 days in vitro) from the rat. The spontaneous release of taurine from both cell types was slow. The release from both neurons and astrocytes was significantly enhanced by 0.1 mM veratridine, the stimulatory effect being more pronounced in granule cells than in astrocytes. No homo or heteroexchange with extracellularly added taurine or its structural analogues could be detected, suggesting that the efflux is probably not mediated via the membrane transport sites. Kainate stimulated the release more from granule cells than from astrocytes, the effect apparently being mediated by kainate-sensitive receptors. Depolarization of cell membranes by 50 mM K+ induced co-release of endogenous taurine and glutamate from both cell types. Preloaded [3H]taurine was readily released from astrocytes by potassium stimulation. Stimulated release occurred from granule cells if they had been cultured for 4 days with the label but not from the cells preloaded for only 15 min. PMID- 2566957 TI - cis-unsaturated fatty acids stimulate catecholamine secretion, tyrosine hydroxylase and protein kinase C in adrenal medullary cells. AB - In digitonin-permeabilized bovine adrenal medullary cells, arachidonic acid and oleic acid, the cis-unsaturated fatty acids, enhanced Ca2+-induced secretion of catecholamines, whereas elaidic acid, a trans-unsaturated fatty acid and stearic acid, a saturated fatty acid, had no effect. Indomethacin, an inhibitor of cyclooxygenase and nordihydroguaiaretic acid, an inhibitor of lipoxygenase, failed to inhibit the stimulatory effect of arachidonic acid. Stimulation of catecholamine secretion by arachidonic acid was abolished by the removal of adenosine 5'-triphosphate and Mg2+ from the incubation medium. Pretreatment of the cells with phorbol 12-myristate 13-acetate, an activator of protein kinase C, enhanced Ca2+-induced catecholamine secretion. In cells pretreated with phorbol 12-myristate 13-acetate, the stimulatory effect of arachidonic acid on Ca2+ induced catecholamine secretion was greatly reduced. In digitonin-permeabilized cells, arachidonic acid and oleic acid enhanced Ca2+-induced activation of tyrosine hydroxylase in the presence of adenosine 5'-triphosphate and Mg2+, whereas elaidic acid and stearic acid did not activate the enzyme. In a soluble fraction of adrenal medullary cells, 32P incorporation to histone by protein kinase C was increased by arachidonic acid and oleic acid, but not by elaidic acid and stearic acid. These results suggest that cis-unsaturated fatty acids modulate Ca2+-induced catecholamine secretion and tyrosine hydroxylase activity by activation of protein kinase C in adrenal medullary cells. PMID- 2566958 TI - [Role of somatostatin and parenteral nutrition in the treatment of acute pancreatitis. Personal experience]. AB - The aim of the study was to evaluate and control the therapeutic validity of Somatostatin administration and the clinical benefits of parenteral nutrition during acute pancreatitis. We selected 31 patients with 1st and 2nd degree pancreatitis according to Ranson's classification. Diagnosis was based on clinical and humoral data and confirmed by echography and CT examinations. The most common etiological cause was biliary++ lithiasis (74.2%). All the patients in the study were split into two groups and received conventional treatment. The therapeutic schedule administered to group 1 included somatostatin (250 micrograms/h for 72-96 h), while group 2 received total parenteral nutrition with 2,000-2,500 Kal/day trough a central vein. The data obtained from our study demonstrated that both somatostatin and parenteral nutrition are valid tools during the acute phase of the disease. It must be pointed out that the former significantly influences the clinical course and allows a precise control of the painful symptomatology, the objective picture and the curve of the main hematochemical parameters. Parenteral nutrition betters the anabolic response of the organism during the acute phase and carries out an indirect antienzymatic response, so favouring a quicker recovery than observed in the group treated with somatostatin. PMID- 2566959 TI - [Medical therapy of peptic ulcer: problems and prospects]. AB - In the last few years, several drugs have been proposed for the healing of peptic ulcers H2 receptor antagonists are probably the reference-drugs for their efficacy and safety. Acute treatment of peptic ulcer with cimetidine, ranitidine or famotidine gives a high healing rate: from 60% to 90%, depending on the location of the ulcer and the drug used. However, relapse after short-term treatment still remains frequent, and prophylaxis of recurrence must be decided. Long-term treatment with antisecretory drugs could be dangerous for changes of the gastric system, with nitrites and nitro-compounds occurrence, and for the risk of frequent relapses. Therefore, the role of mucus-barrier drugs and surgery must be reconsidered. PMID- 2566960 TI - [Growth hormone-releasing hormone. The physiopathologic aspects and its diagnostic-therapeutic use]. AB - The object of the present study is to review all that in the last years has been discovered about growth hormone-releasing hormone (GHRH), in order to point out both its physiopathological characteristics and its possible diagnostic and therapeutic use. In the first section are summarily reviewed the different studies that culminated in 1982 with the identification of three GRF: GRF(1-37) OH, GRF(1-40)-OH and GRF(1-44)-NH2, the last of which, by immunohistochemical methods, resulted to be similar to the hypothalamic hGHRH. Then we describe the anatomic distribution of GHRH in man, and its mechanism of action at both receptor and postreceptor levels. On the other hand, the control of the GHRH secretion by peptidergic hypothalamic neurons occurs through four principal monoaminergic systems such as dopaminergic, noradrenergic, adrenergic and serotoninergic ones, and also by cholinergic fibers and by endogenous opiates, all acting to cause the release, into the hypothalamo-hypophyseal portal circulation, of GHRH. In the second section is attracted attention on the GHRH as a diagnostic agent in the two diseases that represent the main alterations of the GH secretion: acromegaly and short stature. According to the different studies considered, it may be concluded that GHRH testing has limited diagnostic usefulness in the clinical evaluation of acromegaly, but allows to discriminate acromegalic patients with ectopic production of GHRH from those with pituitary tumors. For what concerns short stature, the results of observation realized both in adult subjects and in children, all with GH deficiency, by exogenous administration of GHRH, have pointed out that the majority of the GH deficiency patients have hypothalamic disregulation, and not a pure pituitary deficiency as it has been supposed before GHRH discovery. In the third section is attracted attention on the GHRH as a therapeutic agent. Its possible use in the therapy of the children with GH deficiency is of considerable interest, above all in relation to the hypothalamic pathogenesis of their short stature. PMID- 2566961 TI - [Urinary enzymes: early biochemical indicators in the biological monitoring study of subjects exposed to styrene]. AB - This paper studies the urinary enzymes N-acetyl-beta-glucosaminidase (NAG) and alanine aminopeptidase (AAP) in workers exposed to styrene. It is possible to consider these enzymes as early biochemical indicators of renal damage, owing to their ability to show alterations in the renal tubule when the creatinine clearance is still in the normal range. This preliminary study shows an increase in the two enzymes in workers exposed to styrene. PMID- 2566962 TI - Neuronal bungarotoxin blocks the nicotinic stimulation of endogenous dopamine release from rat striatum. AB - Nicotinic receptors in the brain are receiving increased attention due in part to the recent cloning of receptor subunits and to postmortem studies revealing alterations in receptor density associated with Alzheimer's disease. The peptide neurotoxin neuronal bungarotoxin (NBT) has been shown to block nicotinic cholinergic responses in autonomic ganglia and in retinal ganglion cells. These findings suggest that NBT may be a useful probe for studying nicotinic receptors in the brain. Therefore, we have investigated the effects of NBT on the nicotine mediated enhancement of endogenous dopamine release from rat striatal slices. It was found that the transient increase in dopamine release caused by 100 microM nicotine was completely blocked by 100 nM NBT, indicating that NBT is a functional nicotinic antagonist in this system. PMID- 2566964 TI - Hydrogen sulfide exposure alters the amino acid content in developing rat CNS. AB - Hydrogen sulfide is a widespread environmental pollutant that may produce severe effects on the developing nervous system. Putative amino acid neurotransmitter levels in the rat cerebrum and cerebellum were determined to evaluate the effects of exposure to hydrogen sulfide during perinatal development. The levels of aspartate, GABA, glutamate, glycine and taurine were quantitated using high performance liquid chromatography. With the exception of glycine, all of the amino acids examined were affected by the treatment. On day 21 postnatal, which was the last day of the exposure, aspartate, glutamate and GABA in the cerebrum and aspartate and GABA in the cerebellum were significantly depressed. The observed alterations in the amino acid levels during this critical phase of development may have chronically affected the activity of the neurotransmitters, their receptor sensitivity or their individual target areas. The consequence of one or a combination of such alterations may lead to behavioral and structural abnormalities. PMID- 2566963 TI - NMDA receptor regulation of neuronal morphology in cultured hippocampal neurons. AB - The relationship of the activity of specific neurotransmitter receptors to neuronal morphology was studied in cultured dentate granule neurons from embryonic rat hippocampus. These experiments were made possible by the development of a defined serum-free culture medium for growth at densities low enough to observe isolated neurons. N-Methyl-D-aspartate (NMDA) and its specific non-competitive antagonist, MK801, affected neuronal morphology in a reciprocal fashion. Like glutamate, NMDA stimulated extensive branching of neuronal processes. Not only was 75% of the branching blocked by MK801, but the longest process was 250% longer in the presence of MK801. These results suggest that neurotransmitters that act on the NMDA receptor may contribute to development and synaptogenesis. PMID- 2566965 TI - Generation and characterization of monoclonal antibodies specific for the human neu oncogene product, p185. AB - A series of monoclonal antibodies specific for the extracellular domain of the human neu gene product (p185) have been produced. The generation of these monoclonal antibodies, and their biochemical and immunological characterization is described. The immunization protocol utilized a series of injections of NIH3T3 cells, cyclophosphamide, and a neu transfected NIH3T3 cell line (designated 18-3 7) which expressed the full length human neu-encoded protein. This immunization regimen induced an immune response to the extracellular portion of p185 on the 18 3-7 cells. A panel of ten hybridomas were identified which secreted monoclonal antibodies with a variety of epitope specificities, and reacted with p185 in a number of different experimental formats. As the neu gene product has been associated with human breast cancers, a series of monoclonal antibodies such as these could prove useful in the diagnosis, prognosis and/or treatment of these human malignancies. PMID- 2566966 TI - Spontaneous and evoked quantal neurotransmitter release at the neuromuscular junction of the larval housefly, Musca domestica. AB - The release of neurotransmitter was monitored at the neuromuscular junctions of larval housefly ventrolateral muscles 6a and 7a, using intracellular recording, and a loose patch clamp technique to isolate discrete release sites. Transmitter release occurred spontaneously and could also be evoked by neural stimuli. Spontaneous discharges consisted of events which were randomly distributed in time and of bursts of temporally ordered events. Evoked and spontaneous release occurred in a quantal manner. The quantal content of evoked excitatory postsynaptic currents (EPSCs) was dependent upon the extracellular calcium concentration, increasing with a 3.8 power dependency. The relationship between the quantal content of a response and extracellular calcium concentration was offset by the presence of magnesium in the bathing saline. The rates of decay of miniature EPSCs (mEPSCs) and EPSCs were also found to increase with extracellular calcium concentration, consistent with a non-diffusion limited block of the glutamate receptor-channel complex by calcium ions (KB 2.5 x 10(4) s-1 M-1, P less than 0.01). The frequency of random mEPSCs (0.26 +/- 0.32 Hz, n = 24 cells) was independent of the extracellular calcium concentration. Random mEPSCs were not inhibited by 1 microM tetrodotoxin which blocked mEPSC bursts and neurally evoked responses. EPSCs evoked during mEPSC bursts had a significantly lower quantal content than those EPSCs recorded from the same nerve terminal between bursting, indicating that both of these forms of release recruited quanta from a common pool of transmitter. Following a neurally evoked EPSC the mEPSC frequency was potentiated severalfold, this delayed release was influenced by EPSCs with large quantal contents evoked in saline containing elevated calcium concentrations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2566967 TI - TaqI RFLP at 19q13.1 identified by the anonymous DNA sequence p5B18 [D19S28]. PMID- 2566968 TI - XmnI polymorphism of the human STS gene. PMID- 2566969 TI - A new polymorphism for the dystrophin intragenic probe P20 [DXS269] using BstX1. PMID- 2566970 TI - Two additional TaqI RFLPs in von Willebrand factor gene (VWF) and pseudogene. PMID- 2566971 TI - Stu I RFLP at the human Hep G2/erythrocyte glucose transporter (GLUT) gene locus. PMID- 2566972 TI - A BAM HI RFLP at the human tyrosine aminotransferase (TAT) gene locus at 16q. PMID- 2566974 TI - Detection of familial LPL deficiency by PvuII RFLP. PMID- 2566973 TI - Involvement of a cis-element that binds an H2TF-1/NF kappa B like factor(s) in the virus-induced interferon-beta gene expression. AB - Interferon-beta (IFN-beta) gene is transcriptionally activated following virus infection of various cell types such as fibroblasts. In the previous studies, regulatory DNA sequences that mediate the virus-induced transcriptional activation have been identified within the 5'-flanking region (up to around -117 respect to the CAP site) of the human IFN-beta gene. The sequences contain binding sites (-100 to -61) for a transcriptional activator, IRF-1, the gene of which is also virus-inducible. In the present study, we focused on an additional cis-element, located between the IRF-1 binding sites and TATA box. Interestingly, the element coincides with the previously identified elements for the transcription factors H2TF-1 and NF kappa B. The element, when tandemly repeated, functions in activating the distal gene expression in either constitutive or virus-inducible manner depending on the cell type. The results suggest the importance of cooperation between IRF-1 and H2TF-1/NF kappa B-like factor in the maximal IFN-beta gene induction. PMID- 2566975 TI - The M13 repeat probe detects RFLPs between two strains of the protozoan malaria parasite Plasmodium falciparum. PMID- 2566976 TI - Nsi I RFLP at the X-linked chronic granulomatous disease locus (CYBB). PMID- 2566977 TI - A BglII RFLP demonstrated for the Il-3 gene in normal human blood cells and leukemias. PMID- 2566978 TI - A KpnI polymorphism for the human insulin-responsive glucose transporter gene (GLUT4) on chromosome 17. PMID- 2566979 TI - BamH I restriction fragment alleles of the human T-cell receptor delta (TRD) variable region V2. PMID- 2566980 TI - Identification of the PvuII RFLPs from the switch human immunoglobulin alpha (IGSA) regions. PMID- 2566981 TI - A specific switch alpha probe of the human immunoglobulin IGHA locus. PMID- 2566982 TI - Frequencies of the human immunoglobulin IGHA2*M1 and IGHA2*M2 alleles corresponding to the A2m(1) and A2m(2) allotypes in the French, Lebanese, Tunisian and black African populations. PMID- 2566983 TI - Identification of an Apa I polymorphism within the human adenosine deaminase (ADA) gene. PMID- 2566984 TI - EcoRI RFLP at 19q13.1 identified by the anonymous DNA sequence p58B18 [D19S32]. PMID- 2566985 TI - Assessment and management of postoperative pain in children. AB - This paper focuses on the knowledge base about the assessment and management of postoperative pain in children. The first section deals with the nature and characteristics of postoperative pain. A description of current pain management practices with children, focusing on analgesic administration, is derived from available research literature. These reports suggest that postoperative pediatric pain relief may be inadequate. Recent advances in pain assessment and measurement in all age groups, particularly with verbal children and the new self-report measures, are discussed. The latest developments in pharmacological and nonpharmacological techniques for the relief of children's postoperative pain are also described. Finally, the paper concludes with a few suggestions for pediatricians relative to their role in assisting in the search for better assessment and management techniques in the care of postoperative children. PMID- 2566987 TI - Pharmacologic management of hypertension. New guidelines based on latest studies. AB - The fourth report of the Joint National Committee on Detection, Evaluation, and Treatment of High Blood Pressure varies from the Committee's earlier reports in a number of ways. The introduction of additional effective antihypertensive agents with varying pharmacologic properties has undoubtedly stimulated the Joint National Committee to recommend a more flexible individualized approach to treatment. Diuretics, beta-adrenergic receptor blockers, angiotensin-converting enzyme inhibitors, and calcium channel blockers are all recommended as reasonable first-step agents in treatment of hypertension. The pharmacologic properties, cost, and side-effect profiles of these drugs differ considerably, and the effectiveness of certain agents appears to vary according to the patient's age and race. All these factors should be considered, as should the existence of any concomitant disease, in choosing initial treatment. Whatever drug is chosen, the physician must continue to provide careful follow-up to ensure that adequate blood pressure control is achieved with minimal side effects. PMID- 2566986 TI - Clinical assessment and pharmacologic treatment of pain in children: cancer as a model for the management of chronic or persistent pain. AB - This paper describes methods of assessment and pharmacologic management for chronic or persistent pain in children. This report will be a clinical one, derived from the experiences of a pediatric oncologist (P.M.Z.) and a pediatric pain consultant (L.K.Z.). Case reports will illustrate clinical assessment and management strategies. PMID- 2566988 TI - Histamine H2-receptor blocking activity of ranitidine analogues containing 3 amino-4-alkylaminofurazan moieties. PMID- 2566989 TI - Some aspects of the neurotransmitter control of anterior pituitary function. AB - It is now indisputably established that a cohort of neurotransmitters and neuropeptides affect the secretion of anterior pituitary hormones by modulating the functional activity of specific hypophysiotropic regulatory hormones. Some salient aspects of the role of gamma-aminobutyric acid in the control of prolactin secretion and of catecholamines and acetylcholine in the control of growth hormone secretion are reviewed, also underlying some diagnostic and therapeutic applications of the new findings in neuroendocrine disorders. PMID- 2566990 TI - Pharmacotherapy of coronary insufficiency. PMID- 2566991 TI - The levels of somatostatin in the brain and CSF of rat after carbamazepine administration. AB - Previous studies have suggested that carbamazepine (CBZ), a potent antiepileptic drug, affects the somatostatinergic system in humans and animals; but the results have been contradictory. In the present study we further evaluated the effect of CBZ administration on somatostatin-like immunoreactivity (SLI) in cisternal cerebrospinal fluid (CSF) and in different areas of the rat brain. Somatostatin receptor binding in the cortex of CBZ-treated rats was also studied. Two hours after administration of CBZ at a dose of 30 mg/kg intraperitoneally, which resulted in a serum CBZ concentration of 64 microM, the SLI in CSF was lower than in vehichle-injected controls (P = 0.024, MANOVA). In the hippocampus SLI was elevated to 132% that of vehicle-injected controls (P = 0.016, Mann-Whitney U test). At a dose of 15 mg/kg a slight decrease in SLI was seen in CSF compared to vehicle-injected controls (P = 0.034, MANOVA) but no change was observed in the hippocampus. After administration of CBZ for 7 days (30 mg/kg intraperitoneally twice a day) we were not able to demonstrate any definitive change in SLI of rat CSF (MANOVA). In these rats the SLI in the hypothalamus was elevated compared to vehicle-injected controls (132%, P = 0.016, Mann-Whitney U-test). In experiments with both acute and chronic administration of CBZ, the somatostatin receptor binding was unchanged. The present study suggests that administration of CBZ only slightly affects the somatostatinergic system in the rat brain. PMID- 2566992 TI - The electrically-stimulated spinal reflex in pithed rats: a possible test model for evaluating blockade of central dopamine D1 and D2 receptors. AB - Intravenous injection of the dopamine (DA) D1 receptor agonist SK&F 38393 (4.3 mumol/kg = 1.25 mg/kg), or the DA D2 receptor agonist pergolide (3.2 mumol/kg = 1.25 mg/kg) increased the electrically-stimulated spinal reflex in pithed rats by more than 600 per cent. The specific DA D1 receptor antagonist SCH 23390 potently inhibited the SK&F 38393-induced spinal reflex but not the pergolide-induced reflex. The DA D2 receptor antagonists clebopride and YM 09151-2 inhibited the pergolide-induced reflex only. Two mixed DA D1/D2 antagonists (cis(Z)-flupentixol and zuclopenthixol) inhibited the effects of both SK&F 38393 and pergolide on the spinal reflex, while the neuroleptically inactive isomer of clopenthixol (trans(E)-clopenthixol) was also inactive in this context. Various antagonists (prazosin (alpha 1), idazoxan (alpha 2), 1- propranolol (beta), bicuculline (GABA] were inactive in the test model. The 5-HT2 receptor antagonists altanserin and ketanserin also showed antagonistic effect. It is concluded that the electrically-stimulated spinal reflex in pithed rats can be used as a test model to estimate the blockade of central DA D1 and DA D2 receptors without influence from alpha 1-adrenergic, alpha 2-adrenergic, beta-adrenergic and GABA-ergic receptors. However, a serotonergic receptor antagonism does influence the specificity of the test model. PMID- 2566994 TI - [Early isolated ophthalmological relapse in a case of periarteritis nodosa related to hepatitis B virus]. PMID- 2566993 TI - Comparison of anaesthetic and kinetic properties of thiobutabarbital, butabarbital and hexobarbital after intravenous threshold doses in the male rat. AB - Due to the exceptionally long duration of action of thiobutbarbital the anaesthetic properties of this barbiturate was reinvestigated with an intravenous threshold technique using butabarbital and hexobarbital as references. Adult male rats were used. The criterion of anaesthesia was a burst suppression in the EEG of 1 sec. or more (the "silent second" = SS). The dose which induced the criterion was used as a threshold. The barbiturates were infused with different rates to obtain dose rate curves. After induction of the threshold criterion the animals were either killed and different tissue concentrations were analyzed with a HPLC method or allowed to survive and duration of SS and duration of loss of righting reflex were recorded. With hexobarbital, duration of SS and of loss of righting reflex increased significantly with increasing dose rate. With increasing rates of thiobutabarbital and butabarbital there was in both cases a stepwise increase in duration of SS. At sacrifice, after induction of SS with slow rates brain concentrations of both thiobutabarbital and butabarbital were lower than values recorded after higher rates. The change between the two concentrations was abrupt and occurred at a rate of 20 mg/kg/min. with thiobutabarbital and at the rate of 1.25 mg/kg/min. with butabarbital. This phenomenon was the reverse of acute tolerance which was recorded with hexobarbital and can thus be denoted acute supersensitivity. A kinetic analysis of serum, muscle and fat indicated considerable differences between the barbiturates. As indicated by mortality figures the induction of acute supersensitivity could be potentially dangerous. PMID- 2566995 TI - Cloning and molecular characterization of the trithorax locus of Drosophila melanogaster. AB - The trithorax (trx) locus of Drosophila melanogaster affects segment determination primarily in the thoracic region. Mutant flies show transformations of the third and, to a lesser extent, first thoracic segment toward the second thoracic segment; abdominal transformations also occur. Prior genetic evidence suggested that these effects are based on interactions between trx and genes of the bithorax complex and Antennapedia complex. Further, interactions between the maternal effect locus female sterile homeotic (fsh) and trx have been observed. To aid in a molecular analysis of trx function, we have cloned the locus by a P element transposon tagging approach. Five insertion mutations have been mapped within a region of about 10 kilobases; one of these mutations reverted coincident with the loss of the insertion. Transcription mapping suggests that two RNAs of about 12 and 15 kilobases are the major transcripts of the trx locus and that the transcription unit comprises a region of about 25 kilobases. Transcripts from the trx locus are distributed uniformly in early embryos, but at 14-16 hr after fertilization the ventral nerve cord contains a higher concentration of trx RNA than other regions of the embryo. PMID- 2566996 TI - Loss of heterozygosity for the short arm of chromosome 1 in human neuroblastomas: correlation with N-myc amplification. AB - Partial monosomy of the short arm of chromosome 1 is the most consistent cytogenetic abnormality found in human neuroblastomas, but its overall frequency and significance are unclear. Using a panel of chromosome-1-specific DNA probes that identify restriction fragment length polymorphisms, we demonstrate that 13 of 47 human neuroblastomas (28%) have somatic loss of heterozygosity (LOH) at one or more loci on the distal short arm of chromosome 1. the chromosomal region that shows LOH most consistently is between 1p36.1 and 1p36.3; loss of a gene or genes in this region may be critical for the development or progression of neuroblastomas. The region of LOH in human neuroblastoma may resemble that described for pheochromocytoma, medullary thyroid carcinoma, and melanoma, which are also tumors of neural-crest origin. Although LOH for distal chromosome 1p can occur in early stages of neuroblastoma, the loss usually occurs in tumors of advanced clinical stages. LOH for the short arm of chromosome 1 correlates significantly with N-myc amplification, suggesting that these two genetic events are related. Indeed, these two lesions appear to characterize a genetically distinct subset of particularly aggressive neuroblastomas. PMID- 2566997 TI - Human umbilical cord blood as a potential source of transplantable hematopoietic stem/progenitor cells. AB - The purpose of this study was to evaluate human umbilical cord blood as an alternative to bone marrow in the provision of transplantable stem/progenitor cells for hematopoietic reconstitution. Although no direct quantitative assay for human hematopoietic repopulating cells is at present available, the granulocyte macrophage progenitor cell (CFU-GM) assay has been used with success as a valid indicator of engrafting capability. We examined greater than 100 collections of human umbilical cord blood for their content of nucleated cells and granulocyte macrophage, erythroid (BFU-E), and multipotential (CFU-GEMM) progenitor cells, in many cases both before and after cryopreservation. First it was determined that granulocyte-macrophage, erythroid, and multipotential progenitor cells remained functionally viable in cord blood untreated except for addition of anticoagulant for at least 3 days at 4 degrees C or 25 degrees C (room temperature), though not at 37 degrees C, implying that these cells could be satisfactorily studied and used or cryopreserved for therapy after transport of cord blood by overnight air freight carriage from a remote obstetrical service. Granulocyte-macrophage progenitor cells from cord blood so received responded normally to stimulation by purified recombinant preparations of granulocyte-macrophage, granulocyte, and macrophage colony-stimulating factors and interleukin 3. The salient finding, based on analysis of 101 cord blood collections, is that the numbers of progenitor cells present in the low-density (less than 1.077 gm/ml) fraction after Ficoll/Hypaque separation typically fell within the range that has been reported for successful engraftment by bone marrow cells. Another observation of practical importance is that procedures to remove erythrocytes or granulocytes prior to freezing, and washing of thawed cells before plating, entailed large losses of progenitor cells, the yield of unwashed progenitor cells from unfractionated cord blood being many times greater. The provisional inference is that human umbilical cord blood from a single individual is typically a sufficient source of cells for autologous (syngeneic) and for major histocompatibility complex-matched allogeneic hematopoietic reconstitution. PMID- 2566998 TI - Active transport of gamma-aminobutyric acid and glycine into synaptic vesicles. AB - Although gamma-aminobutyric acid (GABA) and glycine are recognized as major amino acid inhibitory neurotransmitters in the central nervous system, their storage is poorly understood. In this study we have characterized vesicular GABA and glycine uptakes in the cerebrum and spinal cord, respectively. We present evidence that GABA and glycine are each taken up into isolated synaptic vesicles in an ATP dependent manner and that the uptake is driven by an electrochemical proton gradient. Uptake for both amino acids exhibited kinetics with low affinity (Km in the millimolar range) similar to vesicular glutamate uptake. The ATP-dependent GABA uptake was not inhibited by the putative amino acid neurotransmitters glycine, taurine, glutamate, or aspartate or by GABA analogs, agonists, and antagonists. Similarly, ATP-dependent glycine uptake was hardly affected by GABA, taurine, glutamate, or aspartate or by glycine analogs or antagonists. The GABA uptake was not affected by chloride, which is in contrast to the uptake of the excitatory neurotransmitter glutamate, whereas the glycine uptake was slightly stimulated by low concentrations of chloride. Tissue distribution studies indicate that the vesicular uptake systems for GABA, glycine, and glutamate are distributed in different proportions in the cerebrum and spinal cord. These results suggest that the vesicular uptake systems for GABA, glycine, and glutamate are distinct from each other. PMID- 2566999 TI - Structural features of the acetyl-CoA carboxylase gene: mechanisms for the generation of mRNAs with 5' end heterogeneity. AB - Acetyl-CoA carboxylase [acetyl-CoA:carbondioxide ligase (ADP-forming), EC 6.4.1.2] is the rate-limiting enzyme in the biogenesis of long-chain fatty acids. We have previously characterized five acetyl-CoA carboxylase mRNA species that differ in their 5' untranslated regions but not in the coding region. We have now characterized the exon-intron structure of the genomic DNA that encodes the 5' untranslated region of the mRNA. Generation of different forms of the mRNA is the result of the selective use of two promoters and differential splicing of five different exons. These five exons contain a total of 645 nucleotides and they are scattered over a 50-kilobase-pair genomic DNA region that we have characterized. PMID- 2567000 TI - Chromatin configuration of the human CD2 gene locus during T-cell development. AB - To investigate the molecular basis for the tissue-specific expression of the human CD2 gene, its chromatin configuration was assessed by determining DNase I hypersensitivity and the degree of methylation during T-cell lineage commitment and development. Tissue-specific DNase I-hypersensitive sites were found within the 5' promoter region and a region 3' of the gene essential for gene expression. DNase I hypersensitivity of the 5' region correlated strictly with transcriptional activity, whereas hypersensitivity of the 3' region correlated with T-cell progenitor activity or lineage commitment but not necessarily with transcription. Hha I and Hpa II sites around the 5' and 3' regions were undermethylated in CD2-expressing T cells but were more extensively methylated in other cell types. These results define likely regulatory elements both upstream and downstream of the CD2 gene that control its tissue-specific expression. Further, they show that the 3' regulatory region adopts an open chromatin configuration prior to lineage commitment and during early stages of T-cell development before the CD2 gene is transcribed. PMID- 2567001 TI - gamma-Aminobutyric acid (GABA)-induced currents of skate Muller (glial) cells are mediated by neuronal-like GABAA receptors. AB - Radial glia (Muller cells) of the vertebrate retina appear to be intimately involved in regulating the actions of amino acid neurotransmitters. One of the amino acids thought to be important in mediating retinal information flow is gamma-aminobutyric acid (GABA). The findings of this study indicate that enzymatically isolated skate Muller cells are depolarized by GABA and the GABAA agonist muscimol and that the actions of these agents are reduced by bicuculline and picrotoxin. Membrane currents induced by GABA under voltage clamp were dose dependent, were associated with an increase in membrane conductance, and showed marked desensitization when the concentration of GABA exceeded 2.5 microM. The responses had a reversal potential close to that calculated for chloride, indicating that the currents were generated by ions passing through channels. These data support the view that skate Muller cells possess functional GABAA receptors. The presence of such receptors on retinal glia may have important implications for the role of Muller cells in maintaining the constancy of the extracellular milieu, for neuron-glia interactions within the retina, and for theories concerning the generation of the electroretinogram. PMID- 2567003 TI - Rapid increase in enzyme and peptide mRNA in sympathetic ganglia after electrical stimulation in humans. AB - Thoracic ganglia in humans were studied after electrical, preganglionic stimulation using in situ hybridization with synthetic oligonucleotide probes against the catecholamine-synthesizing enzymes tyrosine hydroxylase (EC 1.14.16.2) and dopamine beta-hydroxylase (EC 1.14.17.1) and neuropeptide tyrosine. Immunohistochemical analysis was also performed. Following short peroperative stimulation a severalfold increase in all three mRNAs was found in principal ganglion cells, whereas no definite changes could be detected in enzyme or peptide levels with immunohistochemistry. The results suggest a very rapid and sensitive regulation of genes involved in signal transmission in the sympathetic nervous system of humans. Moreover, they indicate that electrical stimulation of neurons and/or pathways combined with in situ hybridization may be used as a method to define neuronal projections by visualizing increases in mRNAs for transmitter enzymes and/or peptide in target cells. PMID- 2567002 TI - Localization of preferential sites of rearrangement within the BCR gene in Philadelphia chromosome-positive acute lymphoblastic leukemia. AB - The Philadelphia chromosome associated with acute lymphoblastic leukemia (ALL) has been linked to a hybrid BCR/ABL protein product that differs from that found in chronic myelogenous leukemia. This implies that the molecular structures of the two chromosomal translocations also differ. Localization of translocation breakpoints in Philadelphia chromosome-positive ALL has been impeded due to the only partial characterization of the BCR locus. We have isolated the entire 130 kilobase BCR genomic locus from a human cosmid library. A series of five single copy genomic probes from the 70-kilobase first intron of BCR were used to localize rearrangements in 8 of 10 Philadelphia chromosome-positive ALLs. We have demonstrated that these breakpoints are all located at the 3' end of the intron around an unusual restriction fragment length polymorphism caused by deletion of a 1-kilobase fragment containing Alu family reiterated sequences. This clustering is unexpected in light of previous theories of rearrangement in Philadelphia chromosome-positive chronic myelogenous leukemia that would have predicted a random dispersion of breakpoints in the first intron in Philadelphia chromosome positive ALL. The proximity of the translocation breakpoints to this constitutive deletion may indicate shared mechanisms of rearrangement or that such polymorphisms mark areas of the genome prone to recombination. PMID- 2567004 TI - Pertussis in England and Wales: an investigation of transmission dynamics and control by mass vaccination. AB - The epidemiology of pertussis and its prospects for control by mass vaccination in England and Wales are investigated by analyses of longitudinal records on incidence and vaccine uptake, and horizontal data on age-stratified case reports. Mathematical models of the transmission dynamics of the infection that incorporate loss of natural and vaccine-induced immunity plus variable vaccine efficacy are developed, and their predictions compared with observed trends. Analyses of case reports reveal that the individual force of infection is age dependent, with peak transmission in the 5- to 10-year-old age class. A model incorporating this age dependency, along with partial vaccine efficacy and loss of vaccine-induced immunity, generates predicted patterns that best mirror observed trends since mass vaccination was inaugurated in 1957 in England and Wales. Model projections accurately mirror the failure of mass vaccination to increase the inter-epidemic period of the infection (three years) over that pertaining before control. The analysis suggests that this is due to the impact of partial vaccine efficacy. Projected trends do not accurately reflect the low levels of pertussis incidence reported between epidemics in the periods of high vaccine uptake. This is thought to arise from a combination of factors, including loss of natural and vaccine induced immunity, biases in case reporting (where reporting efficiency is positively associated with the incidence of pertussis), and seasonal variations in transmission. Model predictions suggest that the vaccination of 88% of each birth cohort before the age of 1 year will eliminate bacterial transmission, provided the vaccine confers lifelong protection against infection. If vaccine-induced immunity is significantly less than lifelong (or if vaccination fails to protect all its recipients) repeated cohort immunization is predicted to be necessary to eliminate transmission. Future research needs are discussed, and emphasis is placed on the need for more refined data on vaccine efficacy, the duration of natural and vaccine-induced immunity and the incidence of clinical pertussis and subclinical infections (perhaps by the development of reliable serological tests). Future mathematical models will need especially to incorporate seasonality in transmission. PMID- 2567005 TI - Steps in the development of chemical and electrical synapses by pairs of identified leech neurons in culture. AB - Experiments have been made to follow the development of chemical and electrical transmission between pairs of leech neurons in culture. 1. The cell bodies of identified neurons were isolated from the CNS by suction after mild enzyme treatment, together with a length of the initial segment (or 'stump'). The neurons tested were Retzius cells (R), annulus erector motoneurons (AE), Anterior pagoda cells (AP) and pressure sensory cells (P). Pairs of cells were placed together in various configurations, with different sites on their surfaces making contact. 2. When pairs of Retzius cells were apposed with their stumps touching, serotonergic, chemically mediated synaptic transmission became apparent before electrical transmission. By 2.5 h impulses in either of the two Retzius cells produced hyperpolarizing inhibitory potentials in the other. These potentials were reversed by raised intracellular Cl and showed clear facilitation. The strength of chemical transmission between Retzius cells increased over the next 72 h. 3. After chemical transmission had been established, weak non-rectifying electrical transmission became apparent between Retzius cells at about 24-72 h. By 4 days coupling became stronger and tended to obscure chemically evoked synaptic potentials. 4. When pairs of Retzius cells were aligned in culture with the tip of one cell stump touching the soma of the other, chemical transmission also developed rapidly. Transmission was, however, in one direction, from stump to soma. At later stages non-rectifying electrical coupling developed as with stump-stump configuration. With the cell bodies of two Retzius cells apposed, electrical coupling developed after several days, before chemical transmission could be observed. 5. When Retzius and P cells were cultured with their stumps in contact, inhibitory chemical synaptic transmission developed within 24 h. Transmission was always in one direction, from Retzius to P cell. Electrical coupling of Retzius and P cells never occurred whatever the spatial relations of the cells to one another. 6. Annulus erector motoneurons, which contain ACh and a peptide resembling FMRFamide, first developed electrical coupling when the two stumps were in contact and then, later, bi-directional chemical transmission. Anterior Pagoda pairs placed stump-to-stump showed electrical connections. 7. Electronmicrographs revealed the presence of synaptic structures within 24 h after Retzius-Retzius, Retzius-P or AE-AE stumps were apposed. 8. The specificity of connections between cultured cells was similar to that observed in earlier experiments.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2567006 TI - [Hospital standards and professional profiles]. PMID- 2567007 TI - [International congress of nurses 1989. "Health in the schools"]. PMID- 2567008 TI - Computer modeling of membrane-anchored cellular and viral proteins: organization and function. AB - 1. Although secondary structure predictions are only 60% correct, the computer programs provide some idea as to peptide conformation. 2. The detection of hydrophobic domains in membrane-bound proteins allowed the identification of membrane insertional domains in the polypeptide. 3. A combination of computer analyses of protein conformation and the identification of functional domains in polypeptides must be substantiated by experimental data. 4. Computer analyses of membrane proteins may provide a theoretical approach to the selection and development of antibiotics capable of interfering with the insertion and function of viral proteins in cellular membranes. PMID- 2567009 TI - Computer-aided molecular modelling applied to the dopaminergic system. PMID- 2567010 TI - Beta-adrenoceptor modeling based on MEP studies. PMID- 2567011 TI - Binding of agonists and antagonists to beta-adrenergic receptor. PMID- 2567012 TI - Structure-activity relationships of H2-receptor histamine antagonists based on quantum chemical methods. PMID- 2567013 TI - Quantitative structure affinity relationships in a series of alpha-2 adrenergic amines using the molecular lipophilicity potential. PMID- 2567014 TI - Comparison between measured and calculated partition coefficients of beta receptor antagonists. PMID- 2567015 TI - [The effect of gamma rays on medazepam in a methanolic solution]. AB - Nine radiolytical products have been identified in a methanolic solution of Medazepam exposed to gamma-irradiation. A decomposition model taking into account these radiolytical products is presented. The formation of the individual radiolytical products can be deduced from analogous reactions of the radicals e sdv., H, CH2OH, OH and HO2/O2- with several substances. Based on this knowledge of the radiochemical behaviour of Medazepam scavenger systems against radiolysis in solution can be developped. PMID- 2567016 TI - Sucrose sham-feeding in the rat after administration of the selective dopamine D2 receptor agonist N-0437, d-amphetamine or cocaine. AB - Drugs which act as agonists at dopamine receptors, or which increase dopamine release (e.g., d-amphetamine, cocaine) are known to reduce food intake. The present experiments investigated, for the first time, the effects of a highly selective dopamine D2 receptor agonist, N-0437 (0.3-3.0 mg/kg, IP), on 5% sucrose sham-feeding in gastric fistulated rats, and compared these results with those of d-amphetamine (0.1-3.0 mg/kg, IP) and cocaine (3.0-10.0 mg/kg, IP). The results showed that sucrose sham-feeding was resistant to the effects of N-0437, even though the D2 agonist dose-relatedly reduced sucrose real-feeding in intact animals. The two psychomotor stimulants, d-amphetamine and cocaine, produced some reductions in sham-feeding, although in the case of the highest dose of d amphetamine, the pronounced reduction in the consumption of sucrose was probably secondary to induced behavioral stereotypy. The results suggest that D2 receptor stimulation may interact with satiety cues to reduce ingestion of sucrose, but that in the absence of potent satiety stimuli D2 receptor stimulation is ineffective. Furthermore, N-0437 appeared not to be equivalent to either d amphetamine or cocaine in their effects to reduce sucrose sham-feeding. PMID- 2567017 TI - The enzymology and molecular biology of the Ca2+-activated photoprotein, aequorin. AB - Aequorin is a bioluminescent protein, isolated from the hydromedusan Aequorea victoria. A recombinant cDNA plasmid (pAEQ1) was shown to encode apoaequorin by detecting photoprotein activity in an extract of an E. coli strain containing pAEQ1 (Prasher et al., 1986, Biochem. Biophys. Res. Comm. 126, 1259-1268). The nucleotide sequence of the pAEQ1 insert has been determined and is shown to differ significantly from the aequorin cDNA (AQ440) isolated by Inouye et al. (1985, Proc. Natl. Acad. Sci. USA 82, 3154-3158). Comparisons of the coding regions of the two cDNAs show there are 52 nucleotide differences, 19 of which are responsible for 18 amino acid replacements. These differences explain the microheterogeneity observed at 17 positions during the sequencing of native apoaequorin. Five aequorin isotypes extracted from Aequorea tissue are observed on 2-dimensional gels and the E. coli-expressed apoaequorin is shown to co migrate with one of these isotypes. The multiple isotypes could be caused by the presence of a multi-gene family since Southern blot analysis of Aequorea DNA suggests the presence of a minimum of four aequorin genes. Immunoblot analysis suggests that purified native aequorin is proteolytically cleaved during its purification from Aequorea. Comparison of the deduced cDNA translations and the protein sequence suggests the loss of seven residues from the amino terminal. Overexpression of the apoaequorin cDNA in E. coli now provides the means of obtaining gram quantities of a single isotype of the protein which can be converted to aequorin in the presence of coelenterate luciferin, oxygen and an appropriate thiol. Proper extraction procedures and a single chromatographic step provides apoaequorin which is greater than 95% homogeneous. PMID- 2567018 TI - A plasmid to visualize and assay termination and antitermination of transcription in Escherichia coli. AB - To facilitate the analysis of termination and antitermination of transcription in prokaryotes, a complex operon has been assembled into the pBR322 replicon, drawing upon natural and synthetic DNA elements. This operon is initiated from a strongly inducible promoter without temperature restraints. It includes a severe transcription terminator and therefore requires antitermination of transcription to express a downstream lacZ reporter gene. Antitermination can be provided by an upstream N-utilization site from phage lambda, working in conjunction with N protein supplied in trans from a compatible plasmid. In this situation, the nusA gene of Salmonella, substituted into the Escherichia coli host, prevents lacZ function, confirming that a good facsimile of lambda's specific antitermination mechanism has been recreated. The nonessential, easily assayed product of this operon, beta-galactosidase, is also screenable by colony color on chromogenic substrate. The plasmid described will therefore serve as a tester for mutations affecting the various aspects of transcription regulation by termination. PMID- 2567019 TI - The significance of subtyping tardive dyskinesia: a study of prevalence and associated factors. AB - The prevalence of tardive dyskinesia (TD) in 137 Nigerian psychiatric patients was 27%. There were no differences in the prevalence rate between patients with affective disorder and those with schizophrenia. There were also no significant differences between the sexes but a trend for the more severe forms of dyskinesia to be commoner in females was noticed. Demographic, clinical and treatment variables were investigated for association with TD and each of its two putative subsyndromes: orofacial and appendicular dyskinesias. Two cases of severe and persistent tardive dystonia, associated with orofacial TD, were seen in two young adults, one with relatively short exposure to neuroleptics. After initial univariate screening, multivariate statistical methods revealed that different factors were associated with each of the two subsyndromes. While length of hospitalization correlated significantly with orofacial dyskinesia, cumulative duration of exposure to high-potency neuroleptics and number of ECTs received were significantly associated with appendicular TD. Neither age nor sex correlated with either of the subsyndromes. The findings confirm and extend previous observations suggesting that these dyskinesias may involve different pathophysiological mechanisms. PMID- 2567020 TI - Ethopharmacology of social conflict and communication: anxiolytics, antidepressants, antipsychotics, and analgesics. Second international meeting of the European Behavioural Pharmacology Society. Athens, Greece, 31 August-3 September 1988. Proceedings. PMID- 2567021 TI - Temporal patterning of ultrasonic distress calls in the adult rat: effects of morphine and benzodiazepines. AB - Opioids and benzodiazepines modulate the ultrasounds that rats emit before being presented with aversive stimulation. Mild, intermittent 10 s electrical tail stimulation induces rats to emit 20-30 kHz ultrasounds before and after the stimulation. Analysis of the temporal parameters of the sounds via a customized computer system reveals them to be emitted in bouts of about five sounds, each sound being separated from the next by 0.2-0.35 s pauses. Morphine decreases the per cent time vocalizing during the pre-stimulation period, with 6 mg/kg being fully suppressive. This effect is reversed by 1 mg/kg naloxone pre-treatment. Chlordiazepoxide and morphine have opposite effects on the temporal structure of the pre-stimulation calls; chlordiazepoxide induces longer bouts with more pulses, and morphine dissolves the bout structure into a series of single pulses. These differential and selective effects of morphine and benzodiazepines on the occurrence and temporal structure of ultrasounds may be relevant to characteristics of different affective expressions. PMID- 2567022 TI - Temporal and sequential patterns of agonistic behavior: effects of alcohol, anxiolytics and psychomotor stimulants. AB - Social and agonistic interactions are composed of a range of species-typical acts, postures, displays and other communicative signals that follow characteristic patterns. Descriptive and analytic methods permit an assessment of the temporal and sequential features of highly probable patterns of agonistic interactions. Analysis of the intervals that separate consecutive attacks by a resident mouse or rat toward an intruder identifies bursts or epochs of attacks. Amphetamine (1.25, 2.5 mg/kg), but not diazepam or alcohol, alters the burst pattern of attack behavior. Higher doses of alcohol, but not diazepam, in either resident male rats or in lactating rats confronting an intruder, reduce the sequences of aggressive acts and postures with high transition probabilities as identified by lag sequential analysis. These results suggest that temporal and sequential patterning mechanisms may be differentially altered by amphetamine- and alcohol-type substances. These neural for many types of behavior. PMID- 2567023 TI - Quantitative and comparative analyses of pro-aggressive actions of benzodiazepines in maternal aggression of rats. AB - The pro-aggressive effects of low doses of benzodiazepines on maternal aggression in rats were studied. Chlordiazepoxide, diazepam, oxazepam and alprazolam produced bell-shaped dose-response curves, with increased aggression at low doses. Only alprazolam significantly reduced aggression at higher doses. A comparison of the drug effects on different aggressive elements revealed that chlordiazepoxide and oxazepam increased the frequency of more elements of the aggressive repertoire than diazepam or alprazolam. Thus, although all benzodiazepine receptor agonists increased aggression, there were significant quantitative differences in their effects. PMID- 2567024 TI - Differences among nine 1,4-benzodiazepines: an ethopharmacological evaluation in mice. AB - The present study explored whether the profiles of action of benzodiazepines on intraspecies conflict behavior in mice are different. The occurrence of seven behavioral elements was observed in aggressive and timid singly-housed male mice treated with drugs in paired interactions with untreated non-aggressive males. At low doses, some benzodiazepines (alprazolam, oxazepam and diazepam) inhibited defenses, escapes, or attacks, but did not reduce other activities (social sniffing, walking, rearing), and actually increased most of them. At comparable doses, other benzodiazepines (flunitrazepam, nitrazepam, clonazepam and chlordiazepoxide) stimulated only social sniffing, but reduced rearing or walking. Further benzodiazepines (triazolam and lorazepam) reduced defenses, escapes and attacks only at doses that suppressed most of the remaining activities as well. Thus, the nine benzodiazepines tested exhibited different profiles of action in the present study. Alprazolam, oxazepam and diazepam appeared least sedative, while triazolam and lorazepam were most sedative. PMID- 2567025 TI - Behavioral effects of alpha 2 adrenoceptor antagonists and their interactions with ethanol in tests of locomotion, exploration and anxiety in mice. AB - The behavioral effects of two highly selective alpha 2-adrenoceptor antagonists, atipamezole and idazoxan, were investigated in mice using the plusmaze test of anxiety and the holeboard test of directed exploration and locomotor activity. No anxiogenic effect, as assessed by the tendency to enter the closed as opposed to open arms of the plusmaze, was noted for either drug at any dose tested. Neither were there any significant effects on locomotor activity or directed exploration (head-dipping) in the holeboard, or total plusmaze arm entries at any dose of either drug. The co-administration of either atipamezole or idazoxan had no effect on either the anxiolytic effect of ethanol (2 g/kg) or its locomotor stimulant effect in the holeboard. Atipamezole (1.0 and 3.0 mg/kg) significantly reversed the ethanol-induced reduction in exploratory head-dipping; a similar trend was seen for idazoxan. There was also a significant potentiation of the ethanol-induced increase in the number of total arm entries made on the plusmaze caused by 1.0 mg/kg (but not 3.0 mg/kg) atipamezole and both 0.3 and 1.0 mg/kg idazoxan. The results suggest that some of the behavioral effects of ethanol can be reversed by alpha 2-adrenoceptor antagonists whilst others are unchanged. PMID- 2567027 TI - Discrepancy in the time course of EMD 23448 induced yawning and reduction of extracellular dopamine. PMID- 2567026 TI - Specific modulation of social memory in rats by cholinomimetic and nootropic drugs, by benzodiazepine inverse agonists, but not by psychostimulants. AB - The recognition of an unfamiliar juvenile rat by an adult rat has been shown to imply short-term memory processes. In this study the effect of various psychotropic drugs on this investigatory behaviour was examined. The procedure was as follows: an unfamiliar juvenile rat was placed in the home cage of an adult rat for 5 min. The time spent by the adult rat in investigating the juvenile was recorded. The adult rat was then immediately treated with vehicle or test compounds, and was again exposed for 5 min to the same juvenile 2 h later. At this time point vehicle-treated rats no longer recognized the juvenile rat, i.e. the time of investigation was similar to that observed during the first presentation. Arecoline (1 and 3 mg/kg IP), physostigmine (0.05 and 0.1 mg/kg SC), RS86 (0.5 mg/IP) and nicotine (0.125 and 0.5 mg/kg IP) reduced in a dose dependent fashion the time spent in investigating the juvenile during the second exposure. This result cannot be attributed to nonspecific effects, since it was not observed when a different juvenile was used for the second exposure. The effect of arecoline was reversed by scopolamine, but not by methylscopolamine. Aniracetam reduced investigatory behaviour at the dose of 50 mg/kg IP. FG 7142 (5 mg/kg IP) and beta-CCM (0.4 mg/kg IP) were also active and their effect was reversed by Ro 15-1788. DL-Amphetamine (0.5 and 1 mg/kg IP), nomifensine (1.25-10 mg/kg IP) and strychnine (0.25 and 0.5 mg/kg IP) were ineffective or reduced this behaviour unspecifically.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567028 TI - A comparison of the effects of diazepam versus several typical and atypical anti depressant drugs in an animal model of anxiety. AB - We examined the anxiolytic effects of a variety of anti-depressant drugs, administered either acutely or chronically, in an animal model of anxiety involving novelty-suppressed feeding in food-deprived rats. Following a single injection of desipramine (10 mg/kg) amitriptyline (10 mg/kg), mianserin (10 mg/kg), fluoxetine (10 mg/kg), buspirone (4 mg/kg), gepirone (4 mg/kg) or nomifensine (10 mg/kg), there was no decrease in the latency to begin eating in the novel environment such as occurred with diazepam (2 mg/kg). In fact, an increased latency was observed for desipramine, amitriptyline, fluoxetine, and nomifensine. In contrast, chronic (21 days) treatment with each of the above mentioned drugs, except nomifensine, significantly reduced the latency to begin eating relative to vehicle controls. These findings suggest that a variety of tricyclic and novel anti-depressant drugs acquire anxiolytic properties following chronic administration. PMID- 2567029 TI - Discriminative stimulus properties of midazolam: comparison with other benzodiazepines. AB - Rats (N = 12) were trained to discriminate midazolam (1 mg/kg, IP) from vehicle in a food reinforced operant conditioning procedure. Midazolam, flunitrazepam, diazepam, chlordiazepoxide and pentobarbital showed dose-dependent substitution for midazolam. Buspirone and Ro 15-1788 did not substitute for midazolam. The midazolam cue was dose-dependently antagonized by Ro 15-1788. In rats (N = 12) trained to discriminate chlordiazepoxide (3 mg/kg, IP) from vehicle midazolam, flunitrazepam, diazepam and chlordiazepoxide substituted completely and dose dependently for chlordiazepoxide. The relative potency of chlordiazepoxide and diazepam was three times less in the midazolam-trained animals than in the chlordiazepoxide-trained animals. Response rate and latency data further support the main finding that the midazolam cue is similar, but not identical to the cue of classical benzodiazepines. PMID- 2567030 TI - Chronic neuroleptic effects on spatial reversal learning in monkeys. AB - Cebus apella monkeys were chronically administered the antipsychotic drug fluphenazine decanoate for periods ranging from 3.5 to 5.5 years. In the present study, four of these monkeys and two controls were tested for cognitive abilities on a spatial learning task, which consisted of an original discrimination and four reversals of that discrimination. No effect of fluphenazine administration was seen in the rate of learning the original discrimination, but the carryover of learning across discrimination reversals was significantly reduced by fluphenazine. After overtraining on the original discrimination, the controls showed the normal difficulty in learning the first reversal. The fluphenazine treated monkeys showed no such disruption. On subsequent reversals, the controls showed continually improving performance, so that on the third and fourth reversals they had near-perfect scores. On the other hand, the fluphenazine treated monkeys showed no change over the four reversals. Unlike normal monkeys, their learning did not improve with practice. Although simple forms of learning seem to be relatively unaffected by chronic fluphenazine administration, more complex learning is disrupted. PMID- 2567031 TI - Role of specific dopamine receptor subtypes in amphetamine discrimination. AB - Biochemical, electrophysiological, and behavioral experiments suggest that the dopamine D-1 and D-2 receptor subtypes functionally interact. In rats trained to discriminate 1.0 mg/kg d-amphetamine, substitution with the D-2 agonist quinpirole (0.1-2.0 mg/kg) produces amphetamine-lever responding, whereas the D-1 agonist SKF 38393 (0.3-10.0 mg/kg) elicits only saline-appropriate responding. Combining either quinpirole (0.05-0.5 mg/kg) or SKF 38393 (0.5-10.0 mg/kg) with 0.3 mg/kg d-amphetamine results in dose-dependent increases in amphetamine-lever responding. Conversely, the D-1 antagonist SCH 23390 (0.02-0.1 mg/kg) antagonizes the discrimination produced by 0.7 mg/kg d-amphetamine. Additional combination studies examined the effect of DA receptor drugs on discrimination when quinpirole is substituted in d-amphetamine trained rats. SKF 38393 (0.5-7.0 mg/kg) fails to increase the amphetamine-appropriate lever response produced by either 0.05 or 0.2 mg/kg quinpirole. Similarly, SCH 23390 (0.01-0.1 mg/kg) fails to antagonize the amphetamine-lever responding produced by either 0.2 or 0.5 mg/kg quinpirole. Haloperidol (0.02-0.2 mg/kg) does antagonize the amphetamine appropriate response produced by quinpirole substitution. The d-amphetamine discrimination studies indicate that stimulating D-2 receptors alone or D-1 receptors in the presence of d-amphetamine yields d-amphetamine-lever responding, and suggests that D-1/D-2 receptors can functionally interact to alter discrimination behavior. Quinpirole substitution, on the other hand, shows an insensitivity to D-1 receptor manipulations. PMID- 2567032 TI - Yawning and suppression of exploration in amphetamine-treated rats, incompatibility with the autoreceptor hypothesis. AB - The hypothesis that yawning and suppression of exploration, induced by low doses of dopamine agonists, are mediated by stimulation of dopamine autoreceptors was tested by studying the influence of amphetamine on these behavioural models and on extracellular levels of dopamine sampled by microdialysis. Behaviour was measured in a holeboard apparatus. A low dose of amphetamine (0.2 mg/kg) caused slight activation of habituated rats. The same dose of amphetamine completely counteracted the decrease in extracellular dopamine levels caused by pergolide (0.02 mg/kg) and, instead, elevated the dopamine levels to 300% above baseline. The same or higher doses of amphetamine (0.5-1.0 mg/kg) did not completely antagonise suppression of exploration or yawning induced by apomorphine (0.05 mg/kg) or pergolide (0.02 mg/kg). The results suggest that both yawning and suppression of exploration induced by low doses of dopamine agonists are not mediated by dopamine autoreceptors, since these behaviours could be elicited when the extracellular levels of dopamine were elevated above baseline. The alternative hypothesis that these behaviours are mediated by sensitive post synaptic receptors is suggested. It was also found that combined treatment with SCH 23390 (0.05 mg/kg) and amphetamine (2 mg/kg) induced yawning, which further supports the new hypothesis. PMID- 2567033 TI - Differential generalization to pentobarbital in rats trained to discriminate lorazepam, chlordiazepoxide, diazepam, or triazolam. AB - In drug discrimination studies benzodiazepine-trained animals have typically responded on the drug lever when tested with barbiturates. In a recent study, greater specificity appeared to be shown when lorazepam was used as a training drug. The generality and limits of this finding were explored in the present set of experiments. The asymmetrical cross-generalization found in lorazepam- and pentobarbital-trained baboons was replicated in rats and was shown not to be a function of either lorazepam (0.1., 0.32, or 1.0 mg/kg) or pentobarbital (10 or 25 mg/kg) training dose (i.e., pentobarbital-trained rats responded on the drug lever in tests with lorazepam, but lorazepam-trained rats did not show comparable pentobarbital generalization). In the next experiment, groups of rats were trained to discriminate chlordiazepoxide (10 mg/kg), triazolam (0.1 mg/kg), or diazepam (1.0 mg/kg). Generalization to both lorazepam and pentobarbital was shown by these rats. Finally after daily pentobarbital administration, lorazepam trained rats made a sufficient number of responses after high pentobarbital doses to permit extension of the range of pentobarbital doses tested. Pentobarbital generalization increased, but still did not occur in all rats and was unreliable in successive tests in the same rats. These results suggest less homogeneity in the discriminative stimulus effects of "depressant drugs" than generally has been recognized. PMID- 2567034 TI - Effects of benzodiazepine and GABA antagonists on anticonflict effects of antianxiety drugs injected into the rat amygdala in a water-lick suppression test. AB - In order to elucidate the role of the amygdala in rat conflict behavior in a water lick suppression test, we examined the effect of lesions of various nuclei of the amygdaloid complex on this behavior. An anticonflict effect was produced by a lesion of the anterior part of central and basolateral amygdala, and lesion to the posterior part of the central amygdala, but not by posterior of the basolateral amygdala or medial amygdala lesions. These results suggest that the amygdala, especially the anterior part of the central and basolateral nuclei, plays an important role in conflicting behavior of rats in the water lick test. In a second experiment, the effects of benzodiazepine- and GABA-antagonists on the anticonflict action of diazepam, zopiclone, and phenobarbital injected into the anterior part of central and basolateral amygdala were examined, also using a water lick suppression test. A dose-dependent anticonflict action was produced by systemic administration as well as by intra-amygdala injection of diazepam, zopiclone, lormetazepam, flurazepam and phenobarbital. The order of potency was lormetazepam greater than zopiclone greater than or equal to diazepam greater than flurazepam greater than or equal to phenobarbital for both routes of injection. The anticonflict effects of diazepam and zopiclone injected into the amygdala were completely reversed by Ro15-1788 and beta-CCM but not by bicuculline, while the anticonflict effect of phenobarbital was reversed by beta CCM but not by Ro15-1788 or bicuculline. The present results strongly suggest that the anterior nuclei of central and basolateral amygdala are important sites of action of antianxiety drugs, and that an anticonflict action produced by intra amygdala injection of benzodiazepines or barbiturate is mediated through the different receptor mechanisms. PMID- 2567036 TI - New findings with anxiolytic drugs. Proceedings of a satellite conference held during the XVIth Collegium Internationale Neuro-Psychopharmacologicum (C.I.N.P) Congress. Munich, August 15-19, 1988. PMID- 2567035 TI - The relationship of symptomatology and medication to electrodermal activity in schizophrenia. AB - An electrodermal assessment procedure using mild innocuous tones, task-relevant tones, and loud white noise was administered to a sample of schizophrenic patients (n = 77). The present study investigated the relationship of schizophrenic symptomatology and anticholinergic level of neuroleptic medication to electrodermal variables. Each patient was evaluated for positive (florid) and negative (defect) symptoms. The neuroleptics that patients received were rated according to their anticholinergic effect. No significant differences in symptomatology were observed between electrodermal responders and nonresponders; however, nonresponders showed a nonsignificant tendency to have more positive and negative symptoms than responders. Both negative symptoms and anticholinergic levels were significant predictors of lower tonic levels of skin conductance. Patients receiving high anticholinergic neuroleptics showed significantly reduced measures of electrodermal responsivity and tonic levels. Thus, both symptomatology and type of neuroleptic are related to electrodermal measures, with the anticholinergic effect of neuroleptic medication being more pronounced. PMID- 2567037 TI - The new generation of serotonergic anxiolytics: possible clinical roles. AB - Serotonin has been implicated in mediating diverse physiologic and psychologic processes. The anatomy and complex pharmacology of brain-serotonin systems enables this neurotransmitter to broadly affect normal and abnormal behaviors. It appears that serotonin plays a role in multiple psychopathologies, including anxiety, depression, mood disorders, aggressive acting out, alcohol-related syndromes, and disinhibitory disorders characterized by impulsivity. It would not be surprising, therefore, if drugs that alter the dynamics of serotonergic neurotransmission prove to be effective in multiple clinical settings. Such agents may treat broad symptom clusters common to multiple nosologic categories. The new generation of serotonergic anxiolytics, including buspirone, gepirone, ipsapirone, and SM-3997, which interact potently with 5-hydroxytryptamine-1A receptors, may prove to be such symptom cluster drugs. There is a scientific rationale for exploring the clinical utility of these agents in anxiety, depression, mood disorders, aggressive syndromes, and alcohol-related disorders. PMID- 2567038 TI - Animal models for anxiety and response to serotonergic drugs. AB - In examining anxiety and the response of animal models to serotonergic drugs, four aspects should be taken into account: (1) the serotonin receptor is subdivided into at least six receptor subtypes; (2) benzodiazepines have acute anxiety-relieving effects, whereas antidepressants, serotonin-uptake inhibitors, buspirone, and serotonin antagonists have antianxiety effects only after prolonged administration; (3) diagnostic criteria differentiate several distinguishable anxiety disorders that have different responsiveness to serotonin related drugs, and (4) various types of animal models exist, each responding differently to serotonin-related drugs. Perhaps particular animal models are relevant only for the study of one particular type of anxiety disorder. This differentiated view will be used when discussing the role of 5-hydroxytryptamine (5-HT) receptor subtypes in anxiety disorders and anxiety models. The 5-HT1A receptor is implicated in anxiety by the compounds buspirone and 8-hydroxy-2-(di n-propyl-aminotetraline) (OHDPAT). The 5-HT1B or the 5-HT1D receptors play a role in the 'defensive burying' anxiety model and probably mediate antidepressant and antianxiety effects of serotonin-uptake inhibitors. The 5-HT1C receptor plays a role in the aversive brain stimulation anxiety model and could play a role in antianxiety effects of mianserin. The 5-HT2 receptor is selectively blocked by ritanserin. In animal 'conflict models' for anxiety, 5-HT-2-receptor antagonists are active, although they are weaker than the benzodiazepines. The 5-HT3-receptor antagonists are reported to be active in social interaction models for anxiety; however, clinical experience in anxiety using these compounds is not yet available. PMID- 2567039 TI - Serotonin-1A anxiolytics: an overview. AB - The selective serotonin-1A receptor partial agonist anxiolytics represent a new class of pharmacologic agents that have demonstrated efficacy in the treatment of generalized anxiety disorder (GAD). These compounds offer a completely different pharmacologic approach to this disorder from previous medications. The selective 5-hydroxytryptamine-1A (5-HT1A) anxiolytics buspirone, gepirone, ipsapirone, and SM-3997 have several important new and unique features that will be reviewed in this paper. These features include no cross-tolerance with alcohol or benzodiazepines, no evidence of abuse or misuse potential, and no withdrawal symptoms or rebound anxiety on cessation of therapy. The 5-HT1A anxiolytics have no muscle relaxant, sedative, or anticonvulsant properties and do not impair psychomotor functioning. They do have a slower onset of effect than standard benzodiazepines-clinical response is usually noted in 1-3 weeks. The side effect profile is quite different from that of the benzodiazepines. It includes gastrointestinal symptoms such as nausea and diarrhea, headache, dizziness, and restlessness. Some patients with GAD who have received chronic (greater than 1 month) benzodiazepine therapy may not respond as well to these compounds initially as will patients with no prior benzodiazepine treatment, especially if the benzodiazepine has been discontinued only recently. These compounds, buspirone in particular, have been shown to have excellent maintenance and prophylactic properties and to be well tolerated with long-term therapy (greater than 3 months). Because of their unique mechanism of action and side effect profile, and no evidence of misuse or abuse potential or interference with mental acuity, these compounds represent a definite advance in the pharmacologic management of GAD. PMID- 2567040 TI - Serotonin and alcohol: interrelationships. AB - Alcoholism is a multifaceted medicosocial problem. Recent literature discusses a common dyad, alcoholism and anxiety. Both disorders are interdigitated with the brain amine serotonin (5-hydroxytryptamine, 5-HT). Direct 5-HT activation reportedly attenuates alcohol consumption, whereas depletion enhances use patterns. Acute alcohol consumption has also been associated with a transient rise, albeit eventual diminished 5-HT turnover. A variety of 5-HT models have confirmed this observation, e.g., reduced platelet 5-HT content, uptake, and cerebrospinal fluid (CSF) 5-hydroxyindoleacetic acid. Such altered characteristics of 5-HT secondary to chronic alcohol use may explain the frequent morbidity of anxiety and/or depression. Acute alcohol consumption is also associated with accumulation of the 5-HT aldehyde derivative 5 hydroxymethtryptoline. Thus, alcohol may induce the in vivo formation of aldehydes, e.g., beta-carbolines, that themselves possess high lipophilicity and psychotropic activity. Future investigation into 5-HT-specific pharmacologic probes in alcoholism will be interesting. Preliminary research has consistently demonstrated that 5-HT-enhancing agents (e.g., zimelidine or fluvoxamine) decrease alcohol consumption, preference, and short-term memory decrements. Thus, 5-HT appears to represent at least one common denominator for a spectrum of behavioral disorders including anxiety and alcoholism. PMID- 2567041 TI - New evidence that L-glutamate is a transmitter at the squid giant synapse. AB - Experiments are described showing unequivocally that transmission at the squid giant synapse can be reversibly blocked by L-glutamate and its agonists kainate, quisqualate and AMPA, though not by NMDA. This effect is presumably brought about by desensitization. The glutamate antagonists cis-2,3-PDA, GAMS and the new quinoxalinediones CNQX and DNQX are also potent reversible blockers. These findings provide new evidence that L-glutamate is a transmitter at the giant synapse and further suggest that the glutamate receptor may be of the non-NMDA type. PMID- 2567042 TI - Microvascular autotransplantation of intra-abdominal testes. AB - In the period between 1981 and 1987, 23 microvascular autotransplantations of intra-abdominal testes were performed on 18 patients at the Royal Manchester Children's Hospital. Nineteen testes [82.6%] were successfully revascularised; however, four [17.4%] became partially or totally atrophic. Early surgery is recommended--well before the age of 2 years, prior to the onset of testicular damage, in order to reduce the risk of infertility. Although the early results are very encouraging, only long-term, postpuberty follow-up regarding both fertility and malignancy can determine the value of testicular autotransplantation. PMID- 2567043 TI - Rare inflammatory and hereditary connective tissue diseases. AB - Polyarteritis nodosa developing during gestation has an extremely grave prognosis. To an uncertain extent, this results from a delay in recognition and therapy. The diagnosis of PAN is complicated by the expanded differential of common conditions associated with pregnancy such as pre-eclampsia and toxemia which can present with similar symptoms and signs. On the other hand, the pregnant woman with known, quiescent disease has a much better prognosis with only one of four women experiencing exacerbation. In women with Behcet's disease, convincing reports of both pregnancy-related flares and remissions involving primarily mucocutaneous manifestations are found in the literature. Gestational exacerbation of the more serious manifestations including chorioretinitis, vasculitis and CNS disease does not appear to be a problem. Also, a significant effect on fetal development or survival is not evident. The pregnant woman with the Marfan syndrome and pre-existing cardiovascular disease, particularly dilatation of the aortic root, has a substantially increased risk of developing a major complication during gestation most commonly aortic aneurysm, dissection, rupture or insufficiency. Echocardiographic determination of the aortic root diameter is prognostic with a decreased risk at a diameter of 40 mm or less. A diameter of greater than 40 to 45 mm constitutes a significant contraindication to pregnancy. All pregnancies in patients with the Marfan syndrome are considered high risk and frequent evaluations and echocardiograms are required. The EDS patient is subject to a wide range of gestational complications resulting from the basic connective tissue defect manifested clinically by hyperextensible skin, joint hypermobility, connective tissue and vascular fragility, and poor wound healing. The most serious complications occur in type I EDS (gravis) and type IV (ecchymotic) and include extensive perineal tears and hematoma after vaginal delivery, uterine prolapse and rupture, difficulty in suturing wounds and controlling hemorrhage after cesarean section, spontaneous rupture of major arteries, and bowel perforation. Management of the pregnant patient with the EDS must be individualized after identification of the particular type. PMID- 2567044 TI - [Beta blocking drugs in chronic obstructive bronchopneumopathies]. AB - The respiratory tolerance can limit the use of beta-blocker medication (beta-) currently indicated in numerous clinical situations, both cardiovascular and also neurological and ophthalmological. Blockade of beta adrenergic receptors is without risk in subjects free of bronchial pathology, but may destabilize underlying airflow obstruction; either presenting as or sustaining bronchospasm in an asthmatic, and increasing bronchial obstruction, in patients suffering from chronic airflow obstruction (BPCO). Now BPCO, and in particular asthma, are easily associated with diseases for which beta- are indicated. In addition the notion of cardioselectivity is relative and dose dependent. IN PRACTICE: beta- are contraindicated in chronic airflow obstruction with bronchial hyper reactivity; in other cases, beta- should be used carefully choosing the most cardioselective products at the lowest effective dose. Whatever the route of administration (in particular eye drops), the beta- are reliable to produce the same harmful effects. There should be strict follow up of treatment and a watch kept for signs of a worsening or a provocation of respiratory problems; conversely the occurrence of such symptomatology in a patient with bronchial pathology should call into question the place of beta-. Finally beta- should be avoided in allergic subjects and (a fortiori) in specific immunotherapy where there is a possibility of anaphylactic shock. PMID- 2567045 TI - HIV-specific cytotoxic T lymphocytes and their possible implications in the future vaccine against AIDS. PMID- 2567046 TI - Harming and protecting responses to HIV. PMID- 2567047 TI - [Psychopharmaceutic agents in pain treatment]. AB - The chronic pain state can be psychologically induced; on the other hand, pain lasting for months and years can lead to psychopathological alterations. In this context one speaks of the pain-sick patient or of the pain disease. Due to the introduction of psychopharmaca in the treatment of severe chronic pain states, significant progress has been made. Particularly the combination of antidepressants with neuroleptics leads to a substantial reduction of pain. Such a therapy allows substantial savings of analgesics. The indication, the dosage, the mechanism of action and the side-effects of neuroleptics and antidepressants as drugs in the service of pain reduction among patients suffering from chronic pain are covered in the paper. PMID- 2567048 TI - [Convulsions caused by psychopharmaceutical agents]. AB - The convulsive effect of psychopharmaca depends on the 'epileptogenic' potency inherent to the substance, the sedation, the individual convulsion-threshold, epileptic disposition, paroxysmal EEG alterations, acute and chronic organic brain syndromes, alcoholism, high dosage, abrupt discontinuation of medication and forced onset of medication. The literature and the experience show convulsions to occur extremely rarely as a result of antidepressant and neuroleptic medication at standard doses. Caution is needed for patients at risk. PMID- 2567049 TI - Sulphasalazine versus penicillamine in the treatment of rheumatoid arthritis. AB - Fifty-four patients with rheumatoid arthritis were randomized to either sulphasalazine or D-penicillamine in order to compare the short- and long-term efficacy of these two agents in the treatment of rheumatoid arthritis. Decisive improvement was observed in both treatment groups over a 1 year period. Side effects were common in both groups and accounted for termination of therapy in 11 patients during the first year. Radiological deterioration was evident in both treatment groups. A trend toward greater radiological deterioration was observed in patients receiving sulphasalazine, but this was not statistically significant. Only 11 of the 38 patients who completed 1 year of therapy were continuing to take the same drug 5 years later. Eight patients were continuing D-penicillamine and three were still taking sulphasalazine. Among the patients who completed 1 year of therapy, treatment was subsequently terminated because of loss of effective disease control in a significantly higher proportion of patients receiving sulphasalazine (P less than 0.01). The radiological data and the latter observations suggest that D-penicillamine may be a more effective agent for long term treatment. PMID- 2567050 TI - [Parkinson's disease: some physiopathologic aspects and clinical practice]. AB - The lesions of Parkinson's disease chiefly involve the dopaminergic nigrostriatal tract and the great subcortical-cortical dopaminergic, noradrenergic, serotoninergic and cholinergic neuron systems. On the basis of these data, one may suggest a mechanism for some motor and mental disorders, as well as the rationale for treatment with L-dopa, the origin of its main side-effects and the classical escape phenomenon observed with this drug. PMID- 2567051 TI - [Antiparkinsonian drugs]. AB - Levodopa (+ dopa decarboxylase inhibitor) is the most active of all drugs used in the treatment of Parkinson's disease. It acts on both akinesia and rigidity and improves the prognosis of the disease by increasing life expectancy. But levodopa also produces late side-effects: it often induces abnormal movements, fluctuations in motor performance, on-off effects, psychotic hallucinations, etc. Since these late side-effects remain difficult to treat, it is always necessary to assess the benefits and risks of the first treatment with levodopa. Anticholinergic drugs, which mainly act on tremor, must be used with caution since they may induce memory alterations and often confusional states in aged parkinsonians. Dopamine agonists are prescribed as adjuvant therapy in the treatment of the late side-effects of levodopa. New drugs (selegiline), new pharmaceutical preparations (sustained release forms), the first treatment of the disease (levodopa alone versus agonists alone versus levodopa + agonists), together with the new pharmacological approaches (brain grafts, drug infusions) are now under clinical evaluation. PMID- 2567052 TI - [Evolution of enzymuria in the course of acute renal insufficiency caused by non steroidal anti-inflammatory agents]. PMID- 2567053 TI - Agonistic effects of anti-CD2 and anti-CD16 antibodies on human natural killer killing. AB - Two monoclonal antibodies (MoAb), 9-1 (anti-CD2) and 3G8 (anti-CD16), were previously shown to enhance the cytotoxic activity of human natural killer (NK) cells. The present study examined the effect of 9-1 and 3G8 with different effector and target cells to determine whether they activate NK cells through a common mechanism. Analysis of purified lymphocyte subpopulations demonstrated that the CD3+CD16+CD3- NK effector cell population is enhanced by both antibodies, while purified CD2+CD16-CD3+ T cells are not activated by either antibody. Although both antibodies enhance killing of K-562 and Daudi, killing of T-cell lines is enhanced by 9-1 and inhibited by 3G8. In contrast, killing of the promyelocytic cell line, U-937 is inhibited by 9-1 and enhanced by 3G8. On NK susceptible cells the pattern of enhancement with 3G8, an IgG1 MoAb, is consistent with the pattern of target cell expression of an Fc receptor, FcR II, known to bind IgG1 antibodies. This suggests that 3G8 may cross-link effector and target cells through CD16 on the effectors and FcR II on these targets. This could activate NK killing by a mechanism similar to antibody-dependent cellular cytotoxicity reactions (ADCC) with the MoAb in the reverse orientation. The failure of 3G8 F(ab')2 fragments to enhance NK killing, further supports the reverse ADCC mechanism of enhancement by 3G8. The pattern of enhancement mediated by 9-1, an IgG3 MoAb, is not correlated with any target cell Fc-receptor known to bind IgG3 MoAb. The effect of 9-1 may result, instead, from its binding to the unique 9-1 epitope on the CD2 molecule involved in CD2-mediated T-cell activation, as previously described. Alternative mechanisms, including activation of NK killing by 9-1 mediated cross-linking of CD2 and CD16 on the effector cells, have also been discussed. PMID- 2567054 TI - Inheritance of a large deletion within the human immunoglobulin heavy chain constant region gene complex and immunological implications. AB - A deletion of the immunoglobulin heavy chain (IgH) pseudo-gamma, gamma-2, gamma 4, epsilon, and alpha-2 constant region gene segments was found to segregate unchanged in three generations of a family. The IgG1 locus on the IgH allele carrying the deletion was expressed to the same extent as its normal counterpart. One individual who was heterozygous for the deletion had an IgG2 deficiency, whereas the four other heterozygous individuals had serum levels of IgG2 and IgG4 within the normal ranges. IgA2 levels were low or below the normal range in all heterozygous individuals. The data indicate that the expression of some Ig isotypes can be decreased by hemizygous deletions, possibly due to a lower probability for switching. PMID- 2567055 TI - Early mortality after 2,902 coronary artery bypass operations. AB - A review of 2,902 coronary artery bypass grafting operations is presented. During the 16-year study period the mean patient age rose from 51 to 59 years and the average number of grafts per patient from 1.5 to 3.0. There were 81 early deaths (2.8%, the most common cause being myocardial infarction (68%). Left main stem stenosis was present in 23 of these 81 patients and depressed left ventricular function in 30. Compared with the survivors, the deceased patients were characterized by higher age, proportionately large numbers of women, combined procedures and reoperations and less use of internal mammary artery grafts. Of the 94 patients aged greater than or equal to 70, 11 died (12%). The perioperative mortality was significantly greater (p less than 0.05) in women than in men (20/457 vs. 61/2445 viz. 4.4% vs. 2.5%). Combined operations were associated with 8.7% (27/311), reoperations with 6% (6/101) and coronary endarterectomy with 5% (4/75) early deaths. In the last year of the study there were three early deaths among 359 patients (0.8%) who underwent primary isolated coronary bypass grafting without endarterectomy. The perioperative risks fell steadily during 16 years, despite rising proportions of older patients, combined procedures, reoperation and coronary endarterectomy. PMID- 2567056 TI - The role of excitatory amino acids and NMDA receptors in traumatic brain injury. AB - Brain injury induced by fluid percussion in rats caused a marked elevation in extracellular glutamate and aspartate adjacent to the trauma site. This increase in excitatory amino acids was related to the severity of the injury and was associated with a reduction in cellular bioenergetic state and intracellular free magnesium. Treatment with the noncompetitive N-methyl-D-aspartate (NMDA) antagonist dextrophan or the competitive antagonist 3-(2-carboxypiperazin-4 yl)propyl-1-phosphonic acid limited the resultant neurological dysfunction; dextrorphan treatment also improved the bioenergetic state after trauma and increased the intracellular free magnesium. Thus, excitatory amino acids contribute to delayed tissue damage after brain trauma; NMDA antagonists may be of benefit in treating acute head injury. PMID- 2567057 TI - New type of receptor found. PMID- 2567058 TI - Polyarteritis nodosa presenting as recurrent pneumonia following splenectomy for hairy-cell leukemia. PMID- 2567059 TI - Mapping of prolactin and tumor necrosis factor-beta genes on human chromosome 6p using lymphoblastoid cell deletion mutants. AB - A collection of human B lymphoblastoid cell lines (LCLs) was used to map two genetic sequences for which polymorphism had not been identified: human prolactin (PRL) and tumor necrosis factor-beta (TNFB). The LCLs have overlapping deletions on chromosome 6p produced by gamma-irradiation of LCL 721. After using two chromosome 6p sequences for which LCL 721 is heterozygous to validate our scanning densitometry (SD) method for inferring gene copy number, SD was used to map TNFB and PRL. TNFB maps to the interval between the C4 complement and HLA-B loci within the MHC on chromosome 6p. PRL lies within the 6p21.3-6p22.2 interval distal to HLA-C. We found that LCL 721 is heterozygous for PRL DNA fragment lengths generated by HpaII but not MspI digestion, indicating that the two copies of PRL in LCL 721 are differentially methylated. This novel methylation RFLP was used to corroborate the region PRL assignment. PMID- 2567060 TI - Long-range structure of H-ras 1-selected transgenomes. AB - We have used chromosome-mediated gene transfer (CMGT) and whole cell fusion to derive human-mouse hybrid cells carrying reduced human chromosomes 11, by selecting for expression of the transforming H-ras 1 oncogene. To realize the full potential of these somatic cell genetic techniques as resources for enriched DNA probe isolation and the fine structure mapping of chromosomes, the nature of any molecular rearrangements that may accompany the process of DNA transfer must be understood. We have analyzed the long-range structure of our transgenomes by pulsed field gel electrophoresis (PFGE) and show here that, whereas during cell fusion several megabase pairs (Mb) of DNA can be transferred intact, multiple rearrangements of DNA accompany CMGT even in transgenomes where other methods of analysis gave no indication of such molecular scrambling. PMID- 2567061 TI - Gastrinoma. New medical and surgical approaches. AB - The clinical presentation of gastrinoma has changed since the original description by Zollinger and Ellison in 1955. Tumors currently found are smaller, extrapancreatic or extraintestinal in location, and frequently occult within lymph nodes. The incidence of hepatic metastases on initial presentation has decreased. In addition, the clinical course of patients with tumor in lymph nodes is benign, suggesting that more patients than were previously thought are now candidates for cure. Improved knowledge of the anatomic location of gastrinomas has enhanced our ability to find and remove them at laparotomy. As a result of these factors, more patients are being cured than ever before, and in the future, cure rate may be even higher. On the basis of these recent advances, the optimal treatment of gastrinoma is surgical excision for cure. PMID- 2567063 TI - [Quantitative characteristics of various endocrine cells of the duodenal bulb in the pre-ulcer conditions and duodenal ulcer]. AB - Endocrine cells of the duodenal bulb producing adrenalin, noradrenalin, serotonin, histamine and beta-endorphine were assayed in 12 patients with preulcer and 39 patients with duodenal ulcer. It is shown that preulcer was mainly characterized by hyperplasia of endocrine cells in the duodenal bulb mucosa, secreting adrenalin and noradrenalin in the presence of the deficiency of beta-endorphine-producing cells. The recurrent duodenal ulcer was associated with a dramatic fall in the number of catecholamine-containing endocrine cells in the duodenal bulb. PMID- 2567062 TI - Wound healing in nude mice: a study on the regulatory role of lymphocytes in fibroplasia. AB - In order to understand the role of T cells in postinjury fibroplasia, we have studied wound healing in congenitally athymic nude mice that lack a normally developed T cell system. Healing of incisional wounds, as assessed by wound breaking strength, was significantly stronger in nude mice compared with normal thymus-bearing animals. This was accompanied by a marked increase in the amount of reparative collagen synthesized at the wound site, as assessed by the hydroxyproline content of subcutaneously implanted sponges. Because nude mice have some extrathymic T cell maturation, we used an anti-Thy-1.2 (30H12) monoclonal antibody to selectively deplete T cells in vivo. Although such treatments impaired wound healing in normal mice, they had no effect on any wound healing parameter in nude mice. In a separate experiment, T cell reconstitution of nude mice, sufficient to significantly enhance in vivo delayed hypersensitivity responses, led to a decrease in both wound breaking strength and hydroxyproline deposition in subcutaneously implanted polyvinyl sponges. The data suggest that T cells play a dual role in wound healing: an early stimulatory role on macrophages, endothelial cells, and fibroblasts, and a late counterregulatory role, which may be responsible for the orderly completion of wound repair. PMID- 2567064 TI - [Morphology and aggregative properties of erythrocytes in patients with hemorrhagic fever with nephrotic syndrome]. AB - Fifty-six patients with hemorrhagic fever and renal syndrome (mainly with grave and medium-grave disease patterns) were examined for the morphology of red blood cells using scanning electron microscopy. In addition, a study was made of the deformability and aggregation properties of red blood cells. The role of the impaired erythrocytic component of hemostasis in the pathogenesis and clinical picture of hemorrhagic fever and renal syndrome is discussed. PMID- 2567065 TI - Limb defects in gray short-tailed opossums (Monodelphis domestica) following postnatal injection with ethanol or saline. AB - Because the hindlimbs of marsupials at birth are in an embryonic stage of development, this group can prove useful for studies of limb teratology. In this study, injection of neonatal gray opossums with 2 mg ethanol (ETOH) or saline (SAL) in the right or left hindquarter resulted in defects of the associated limb in 44% of ETOH animals and 16% of SAL animals. Affected SAL animals showed gait abnormalities, foot clubbing, and moderately reduced limb size. Affected ETOH animals showed these abnormalities as well as fused digits, missing digits, and, in one case, a partially missing limb. Thus, while injection itself may have resulted in altered limb development, local infusion of alcohol had a further teratogenic effect on such development. PMID- 2567066 TI - The antioxidant butylated hydroxytoluene can retard cerebellar degeneration induced transplacentally by a single low dosage of N-methyl-N-nitrosourea. AB - Late-onset cerebellar degeneration can be induced transplacentally in mice by a single low-dose (1 mg/kg) injection of the direct-acting DNA alkylating agent N methyl-N-nitrosourea (MNU) on day 16 of pregnancy. The offspring develop a mild ataxia that manifests by 12-16 weeks of age postnatally when the animals are challenged with a motor coordination task. Morphological evidence of degeneration includes pyknosis of Purkinje cells and abnormal foliation patterns. Additionally, these animals demonstrate a progressive retinopathy characterized by thinning of the nuclear and plexiform layers of the retina. Efforts to retard the cerebellar degeneration were undertaken in the present study. MNU-exposed and control animals were fed a standard mouse chow diet supplemented with 0.75% butylated hydroxytoluene (BHT), an antioxidant. This supplementation commenced 24 h following exposure to the teratogen and continued throughout the life of the offspring. A second group of MNU-exposed and control mice were fed a non-BHT supplemented, standard Purina mouse chow diet. Quantitative histological evaluation of cerebellar coronal sections indicated that by 4 weeks of age BHT fed, MNU-exposed mice had significantly fewer pyknotic Purkinje cells than non BHT-fed, MNU-exposed animals. This was true for the vermal, paravermal, and lateral areas of the cerebellum. The findings suggest the usefulness of teratogenic models of degenerative diseases for the testing of potential intervention strategies. PMID- 2567067 TI - Effects of in vivo and in vitro exposure to rhodamine dyes on mitochondrial function of mouse embryos. AB - Cationic rhodamines (Rh 123 and Rh 6G) can cause developmental toxicity in mice and inhibit embryonic mitochondrial respiration following in vivo or in vitro dye exposure. Rh B, a neutral rhodamine, fails to show such effects at comparable doses. To assess effects of rhodamines on development, F0F1ATPase activity and ADP translocation were measured on gestation day (GD) 12 in embryonic and adult mitochondria. ATP synthesis in embryonic mitochondria transplacentally exposed to Rh 123 (15 mg/kg/day) or Rh 6G (0.5 mg/kg/day) given to dams by i.p. injection from GD 7 to 10 were inhibited 39% and 49%, respectively. When isolated mitochondria were treated, dose-dependent inhibition was seen; at 5 micrograms of dye/mg mitochondrial protein, ATP synthesis was inhibited 65% and 81% by Rh 123 and Rh 6G, respectively. When F0F1ATPase activity was assessed, in vitro Rh 123 and Rh 6G exposures at levels up to 8 micrograms/mg mitochondrial protein resulted in enzyme inhibition, but at 10 micrograms/mg, ATPase activity was stimulated. Uncoupler-stimulated ATPase activity was also inhibited. ADP translocation was decreased by 19.1% and 37.7% by Rh 123 and Rh 6G, respectively, at dye concentrations of 20 micrograms/mg. Results of in vitro exposure of maternal liver mitochondria were similar to those for embryonic mitochondria, whereas liver from dams exposed in vivo on GD 7-10 was unaffected on GD 12. In vivo or in vitro treatment with Rh B did not affect any embryonic or maternal parameters. Such results support the hypothesis that inhibition of mitochondrial energy metabolism is a mechanism for the developmental toxicity of cationic rhodamines. PMID- 2567069 TI - Beneficial effects of ascorbic acid on preimplantation mouse embryos after exposure to cyclophosphamide in vivo. AB - To study mechanisms of embryotoxicity in early pregnancy, we have evaluated the genotoxic and embryolethal effects of ascorbic acid (AA) alone or in combination with cyclophosphamide (CPA). Female mice were exposed on day 3 of pregnancy. Embryotoxicity was investigated at term and genotoxicity shortly after treatment using the chromosomal aberration test and the sister chromatid exchange (SCE) assay as sensitive end points. Additionally, cytotoxic effects were determined by a proliferation test. AA was not found to be embryotoxic, cytotoxic, or genotoxic when given alone. In combination with 10 mg/kg CPA, however, which induced 50% aberrant metaphases, 100% increase SCE frequency, and a strong inhibition of cell proliferation, AA in a dose range of 25-1,600 mg/kg did not change SCE and proliferation, but reduced the rate of aberrant metaphases significantly. This anticlastogenic effect was clearly correlated to a beneficial effect on embryolethality at term when 200 mg/kg ascorbic acid was given in combination with 40 mg/kg CPA. The results suggest that during early pregnancy AA is not genotoxic even at so-called megadoses doses, but it seems to protect early embryos against damage induced by genotoxic agents like CPA. PMID- 2567068 TI - Interference of prenatal and postnatal exposure to Ca2+-antagonist agents on rat functional development of vascular system. AB - Exposure to drugs during pregnancy can alter functional development of the vascular system. The present investigation was carried out in order to evaluate the effects of prenatal and postnatal exposure to Ca2+-antagonist (diltiazem, verapamil, and nimodipine) drugs on the development of rat vasomotor reactivity. Studies were carried out on pregnant female albino rats exposed from the first day of pregnancy until weaning to diltiazem and verapamil (6 and 24 mg/kg in their drinking water ad libitum) and nimodipine (3 and 12 mg/kg in their food ad libitum). After weaning, pups were exposed until the 60th day of age to the same treatment as their mothers were. Afterwards, pups from the 60th to 90th day of age were fed with a normal diet. In 30-, 60-, and 90-day-old conscious and anaesthetized pups, we evaluated the following: 1) systolic arterial blood pressure; 2) vasomotor responses elicited by various agents: L-noradrenaline (0.1, 1, and 5 micrograms/kg IV), L-isoprenaline (0.01, 0.1, and 1 micrograms/kg IV), and acetylcholine (0.01, 0.1, and 1 micrograms/kg IV) and by sinus-carotid baroreceptor stimulation; and 3) catecholamine, acetylcholinesterase, and adenosinase plasma levels. Prenatal and postnatal exposure to Ca2+-antagonist drugs significantly (P less than .05) decreased the pressor response to sinus carotid baroreceptor stimulation and to L-noradrenaline and increased the hypotensive responses to L-isoprenaline and acetylcholine. Moreover, this type of treatment, although it induced a significant (P less than .05) decrease of catecholamine plasma levels, did not modify the acetylcholinesterase and adenosinase plasma levels in 30- and 60-day-old rats. On the 90th day of age, the evaluated parameters were not different from those of control rats. Our results showed that exposure to Ca2+ antagonists during pregnancy and the postnatal period may alter the functional development of rat vasomotor reactivity. PMID- 2567070 TI - Biochemical effects of three carcinogenic chlorinated methanes in rat liver. AB - Three chlorinated methanes, carbon tetrachloride, chloroform, and methylene chloride, known to cause liver tumors in rodents, were given by oral gavage to adult female rats both 21 h and 4 h before sacrifice. Then hepatic DNA damage, ornithine decarboxylase (ODC), cytochrome P-450, glutathione content, and serum alanine aminotransferase (SGPT) activity assays were performed. Carbon tetrachloride increased rat hepatic ODC activity and decreased cytochrome P-450 content at doses both below and above cytotoxicity (as measured by increased SGPT activity). At 54 and 160 mg/kg, chloroform increased hepatic ODC activity with minimal or no elevation in SGPT activity. At 480 mg/kg chloroform increased hepatic ODC and SGPT activity. A dose of 1,275 mg/kg methylene chloride caused a small, but significant amount of hepatic DNA damage. When these three compounds are compared on either an equimolar or equitoxic (1/5 LD50) basis, their ability to induce hepatic ODC or increase SGPT activity was carbon tetrachloride greater than chloroform greater than methylene chloride. The results of this biochemical study are interpreted with respect to the ability of chemicals to cause hepatic cancer by either genetic or epigenetic mechanisms. PMID- 2567072 TI - [Mammography screening. A statement from the consensus conference on mammography screening, Soria Moria, Oslo, February 8-10, 1989]. PMID- 2567071 TI - Sister chromatid exchange in human chromosomes from normal individuals and epileptic patients on combinations of anticonvulsants. AB - Sister chromatid exchange (SCE) frequency, a sensitive indicator in mutagenicity testing, and mitotic index (MI) have been studied to observe genotoxic effects in epileptic patients on routine combinations of anticonvulsant therapy. All patients, both male and female and from various age groups, revealed an increased frequency of SCE per metaphase and a low MI (P less than 0.001) with respect to controls. A nonsignificant decrease in SCE frequency has been observed with an increase in the age of onset of epilepsy. Although the SCE frequency increased and the MI decreased in some groups with respect to the duration of epilepsy, there was no difference observed in SCE frequency with the duration of therapy. PMID- 2567073 TI - 2-Bromohydroquinone-induced toxicity to rabbit renal proximal tubules: the role of biotransformation, glutathione, and covalent binding. AB - 2-Bromohydroquinone (BHQ) is a model toxic hydroquinone and plays an important role in bromobenzene-induced nephrotoxicity. Proximal tubules isolated to contain decreased glutathione (GSH) levels were at least twice as sensitive to the GSH depleting effects of BHQ and BHQ-induced mitochondrial dysfunction as were tubules with "normal" (i.e., in vivo) GSH content. The decrease in tubular GSH content resulted from BHQ-GSH conjugate formation. A mono-GSH conjugate (2-bromo 3-(glutathion-S-yl)hydroquinone) and a di-GSH conjugate (2-bromo-3,5- or 6 (diglutathion-S-yl)hydroquinone) were identified. In addition, a glucuronide conjugate was identified (2-bromo-1- or 4-O-glucuronylhydroquinone). BHQ-GSH conjugates were not responsible for BHQ-induced toxicity since (1) tubules with normal levels of GSH were more resistant to BHQ-induced toxicity even though they formed more BHQ-GSH conjugates than tubules with decreased GSH levels and (2) inhibition of gamma-glutamyltranspeptidase did not prevent BHQ-induced toxicity. BHQ-equivalents bound covalently to tubular protein in a concentration-, time-, and temperature-dependent manner with the majority of the binding (61%) occurring during the first 15 min after exposure to 0.2 mM BHQ. Tubules pretreated with GSH underwent less BHQ-protein alkylation and mitochondrial dysfunction, and the amount of BHQ recovered and BHQ-di-GSH conjugate formed increased. These data suggest that BHQ is biotransformed to a reactive intermediate (2-bromoquinone and/or 2-bromosemiquinone) and that this intermediate can react with GSH to form BHQ-GSH conjugates and/or bind covalently to tubular protein which may result in mitochondrial dysfunction and tubular death. PMID- 2567074 TI - Chemical modification of equinatoxin II, a lethal and cytolytic toxin from the sea anemone Actinia equina L. AB - The role of arginine and tyrosine in cytolytic properties of equinatoxin II, isolated from the sea anemone Actinia equina L., was studied by means of chemical modifications. The toxin was modified with 2,3 butanedione and tetranitromethane, respectively. The extent of modification and physico-chemical properties of the modified proteins were checked with amino acid analysis, isoelectric focusing and circular dichroic spectra. Extensive treatment of the toxin with 2,3 butanedione modified seven arginines and also two tyrosines, with resulting loss of hemolytic activity. Modification of two out of nine arginine residues resulted in a 25% loss of hemolytic activity, whereas nitration of three out of ten tyrosines decreased hemolytic activity by 95%. The nitrated toxin had at least a 30-fold higher i.v. LD50 than the native toxin. None of the modifications significantly affected the secondary structure of the toxin as revealed by the CD spectra. It is concluded that tyrosine residues are involved in both lethal and cytolytic activity, while the role of arginine residues is not evident because of the non specific alteration of tyrosine residues with 2,3 butanedione. PMID- 2567075 TI - Palytoxin promotes potassium outflow from erythrocytes, HeLa and bovine adrenomedullary cells through its interaction with Na+, K+ -ATPase. AB - Erythrocytes from four mammalian species were compared with regard to K+ loss triggered by palytoxin, to Na+, K+ -ATPase activity, and to ouabain sensitivity of both events. Palytoxin sensitivity (EC50) decreased in the order rat, man (approximately equal to 1 pM) greater than cattle (approximately equal to 500 pM) greater than dog (greater than 10 nM). Na+, K+ -ATPase activity, as measured by Rb uptake, was in the series rat greater than man greater than cattle greater than dog. The glycoside potently inhibited both palytoxin action and ATPase activity in man, cattle and dog erythrocytes, but weakly in those from rats. Ca2+ promoted the palytoxin effects on all erythrocytes. As shown for human erythrocytes, Sr2+ and Ba2+ but not Mg2+ can substitute for Ca2+, and sucrose can substitute for sodium chloride. Human HeLa and bovine adrenomedullary cells also lost their K+ within a few min when exposed to palytoxin (1-10 pM). Ouabain acted as a palytoxin antagonist on both cell types. We conclude that: (a) the ouabain binding site of Na+, K+ -ATPase is part of the palytoxin receptor in every cell type tested, (b) high palytoxin sensitivity is not necessarily accompanied by high ouabain sensitivity, and (c) active ion transport is not a precondition for the action of palytoxin or for its inhibition by ouabain. PMID- 2567076 TI - Stabilization of lethal and hemolytic activities of box jellyfish (Chironex fleckeri) venom. AB - The stability of both the lethal and hemolytic activities of box jellyfish (Chironex fleckeri) tentacle extract was assessed after various extraction procedures. Both activities were higher when no buffers or water were used during the initial extraction. Also, when the extract was first filtered through a Sep pak C18 cartridge, the residual lethal titre, after incubation for 24 hr at room temperature, was increased 16-fold and hemolysis was increased 2.6-fold. Evidence for proteolytic activity in the extract was also obtained and monitored by size exclusion HPLC. PMID- 2567077 TI - Interfacial activity of an ion channel-generating protein cytolysin from the sea anemone Stichodactyla helianthus. AB - The ability of a purified sea anemone (Stichodactyla helianthus) protein cytolysin to interact with a variety of interfaces was investigated by means of the Wilhemy plate method. At the air:water and lipid:water interfaces, the toxin lowered the surface pressure most readily as the aqueous phase pH increased towards the isoelectric point (9.8) of the toxin. The affinity of the toxin for both the phospholipid:water and the oil:water interfaces was much greater than for the air:water interface. Although the toxin had previously been found to avidly bind to sphingomyelin-containing phospholipid dispersions and bilayers, it failed to display any preferential interaction with a sphingomyelin monolayer relative to one of dipalmitoylphosphatidyl-choline under identical conditions, even when the monolayers were maintained at 40 dynes/cm, a pressure considered to produce phospholipid packing densities similar to those observed in cell membranes. Unlike many other membrane-active protein cytolysins, the ability of Stichodactyla cytolysin to penetrate these phospholipid monolayers was not affected by the initial surface pressure over the range 0-32 dynes/cm. However, at 40 dynes/cm initial packing pressure, the surface pressure generated by the cytolysin was similarly reduced in both sphingomyelin and dipalmitoylphosphatidylcholine monolayers. Our results suggest that the protein cytolysin initially binds reversibly to cell and artificial bilayer membranes in a non-specific manner; sphingomyelin domains in the bilayer then provide optimal conditions for insertion into the membrane and subsequent assembly of a stable multimeric complex which functions as an ion channel. PMID- 2567079 TI - [Regulation of tyrosine hydroxylase activity in the rat hypothalamus with gamma aminobutyric acid]. AB - It is established that GABA interacts with tyrosine hydroxylase through the allosteric site which is not identical to sites of tyrosine, DOPA, pterin cofactor, dopamine binding. This interaction is very significant in the GABA influence on the regulation of the tyrosine hydroxylase activity by presynaptic receptors. GABA is supposed to be able to cause dissociation of oligomeric forms of tyrosine hydroxylase. PMID- 2567078 TI - [The All-Union Forum of Physicians]. PMID- 2567080 TI - [Ophthamologic pathology in non-specific aortoarteritis]. AB - Nonspecific aortoarteritis is a systemic autoimmune disease eventuating in gradual stenosis of the aorta and the main vessels with ischemia of the respective organs. Ophthalmologic symptoms have been examined in 54 patients with nonspecific aortoarteritis. Subjective disorders of vision (short-term binocular blindness, metamorphopsia, pain behind the eye, amaurosis fugax) have been detected in 52% of the examinees. Organic lesions of the eye have been diagnosed in 60% of the patients: hypertensive angiopathy (22%), venous stasis retinopathy (17%), occlusion of the central retinal artery (1%), etc. Three possible mechanisms of the development of ocular symptoms have been established: (1) a result of symptomatic hypertension, (2) chronic ocular ischemia, (3) acute hemodynamic ocular circulation insufficiency. PMID- 2567082 TI - [Readers' conference on the work of the editorial board of the journal "Vestnik khirurgii im. I.I. Grekova"]. PMID- 2567081 TI - [Endocrine cells of the internal ear in the normal state and in specific labyrinthine lesion in rabbits]. AB - Using light and electron microscopes, apudocytes of internal ear in normal and syphilis-infected rabbits were examined. Temporal bone blocks were dissected from 16 healthy (control) and 32 infected (experimental) animals. Some of the preparations were hematoxylin and eosin stained for histological examinations; some were silver stained according to Griemelius for histochemical examinations; and others were used to prepare ultrathin sections for electron microscopy. It was demonstrated that apudocytes were normally found in different labyrinthine structures and consistently seen in the vascular stripe. Electron microscopy showed that the vascular stripe included endocrine cells containing secretory granules which are characteristic of serotonin-, melatonin- and catecholamine producing cells. Apudocytes from the syphilis-infected labyrinth were in the state of hyperfunction: the preparations stained according to Griemelius had lowered quantities of apudocytes and contained light cells with the hormone utilized completely; electron microscopy preparations showed granules in the capillary lumen that were transferred there by exocytosis. These observations give evidence that endocrine cells are involved in processes that develop in internal ear both in the norm and pathology. PMID- 2567083 TI - HSV hepatitis in the mouse: a light and electron microscopic study with immunohistology and in situ hybridization. AB - In order to characterize better the morphology and immune response in acute necrotizing HSV infection, murine HSV hepatitis was examined. BALB/c mice were inoculated intraperitoneally with 10(6) plaque-forming units (PFU) of HSV-1 (Lenette) and HSV-2 (D316). In both groups half the animals were pretreated with silica particles to block macrophage function. Up to 6 days after infection four mice from each group were sacrificed at daily intervals and the livers were examined by light and electron microscopy, immunohistology, in situ hybridization, combined immunohistology/in situ hybridization and titration of viral PFU. HSV-2 infected mice developed severe necrotizing hepatitis with persistence of HSV in the liver tissue until the end of the study. HSV-1 infected mice rapidly eliminated the virus and revealed only small necrotic foci. Early phase alterations and necrotic phase lesions were distinguished and characterized and morphologic evidence of a direct cytopathic effect of HSV was detected. A specific immune reaction in late stages appeared to be mediated by T4-positive T lymphocytes. In situ hybridization and immunohistochemistry showed a close correlation with virus titration and were valuable in characterizing early phases and in the assessment of prognosis and differential diagnosis. PMID- 2567084 TI - Proliferation-associated expression of DNA methyltransferase in human embryonic lung cells. AB - The cell cycle-dependent and proliferation-associated expression of the enzyme DNA methyltransferase has been evaluated immunocytochemically in synchronized L 132 human embryonic lung cells, using the anti-DNA methyltransferase monoclonal antibody M1F6D7/5C10. DNA methyltransferase-reactivity was firstly seen in mid-G1 cells. An intense and granular reaction in the cell nuclei with a sparing of the nucleoli was observed in addition to a homogenous and faint cytoplasmic staining. The staining intensity in the cell nuclei increased progressively up to mitosis. In early mitotic cells an intense perichromosomal staining was observed in addition to a homogenous staining of cyto- and karyoplasm after the resolving of the core membrane. In late mitosis the staining intensity decreased rapidly. Early G1 cells and density inhibited, resting G0 cells showed no DNA methyltransferase reactivity at all. Our results indicate that anti-DNA methyltransferase monoclonal antibodies could become valuable tools to detect proliferating cells in cell cultures and tissues. PMID- 2567085 TI - Immunohistochemical analysis of normal and mutated ras oncogene p21 expression in human pulmonary and pleural neoplasms. AB - In this study we examined 214 cases of primary human pulmonary neoplasms for the expression of a mutated form of the ras oncogene p21 product, recognized by the monoclonal antibody (MCA) DWP. Adjacent serial sections from these same cases had previously been used to demonstrate the frequency of ras p21 expression using the broadly reactive anti-ras p21 MCA RAP-5. Confirmation of the increased expression of p21 was accomplished using MCA Y13-259. The use of adjacent tissue sections from these cases allows the direct comparison of the expression of the mutated and non-mutated forms of ras p21. If reactivity with DWP would prove to be significantly more restrictive than that of the "pan" ras MCAs, RAP-5 and Y13 259, it would lend support to the possibility that DWP (and similar MCAs which detect other specific mutations) could be used to define subsets of these neoplasms based on their specific ras p21 phenotype. Since one would anticipate that the valine/cysteine substitution at position 12 of the ras p21 would occur at only low frequencies in human tumors, our results with DWP are consistent with this hypothesis. As previously reported, RAP-5 reacted with a high proportion of lung tumors (100/214 or 47%). In this report, we demonstrate the selective expression of the mutation recognized by the MCA DWP in only 5% of these same tumors (13/214), and that the expression of this mutated form is not restricted to any of the conventional histological subclasses of pulmonary neoplasms. PMID- 2567086 TI - Lysosomal enzyme leakage during the hypoxanthine/xanthine oxidase reaction. AB - Impairment of lysosomal stability due to reactive oxygen species generated during the oxidation of hypoxanthine by xanthine oxidase was studied in rat liver lysosomes isolated in a discontinuous Nycodenz gradient. Production of O2.- and H2O2 during the hypoxanthine/xanthine oxidase reaction occurred for at least 5 min, while lysosomal damage, indicated by the release of N-acetyl-beta glucosaminidase, occurred within 30 s, there being no further damage to these organelles thereafter. The extent of lysosomal enzyme release increased with increasing xanthine oxidase concentration. Superoxide dismutase and catalase did not prevent lysosomal damage during the hypoxanthine/xanthine oxidase reaction. Lysosomes reduced xanthine oxidase activity, as assessed in terms of O2 consumption, only slightly but substantially inhibited in a competitive manner the O2.- -mediated reduction of cytochrome c. This inhibition was almost completely reversed by potassium cyanide, thus pointing to the presence of a cyanide-sensitive superoxide dismutase in the lysosomal fraction. However, potassium cyanide did not affect the hypoxanthine/xanthine oxidase-mediated lysosomal damage, thus suggesting an inability of the lysosomal superoxide dismutase to protect the organelles. Negligible malondialdehyde formation was observed in the lysosomes either during the hypoxanthine/xanthine oxidase reaction or with different selective experimental approaches known to produce lipid peroxidation in other organelles such as microsomes and mitochondria.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567087 TI - An endogenous colon mitosis inhibitor reduces proliferation of colon carcinoma cells (HT 29) in serum-restricted medium. AB - Extracts of mouse intestine contain a colonic epithelial mitosis inhibitor that has recently been purified and identified as a tripeptide (pGlu-His-GlyOH). In order to elucidate further the biological characteristics of this peptide, the effect of the tripeptide on cell proliferation in a human colon carcinoma cell line (HT 29) was examined. The incorporation of tritiated thymidine was significantly reduced at 20-30 h after addition of the tripeptide. The dose response relationship was bell-shaped with loss of inhibitory effect at high or low doses. The number of cells were significantly reduced at a peptide concentration of 10(-8) M at 24 h, but not at 48 or 72 h after addition of the peptide. The inhibition was reversible, and was only observed when the cells were grown in a serum-restricted medium (1%). The inhibitory effect was abolished by increasing the serum content to 10% or adding insulin to the medium. PMID- 2567088 TI - Proliferative status of colonic mucosa in organ culture: 3H-thymidine-labelling studies and computer modelling. AB - 3H-thymidine labelling studies and a computer simulation have been employed to assess proliferative status and cellular organisation in colonic explants maintained in culture for 5 to 7 days. The one-hour flash labelling index (Is) for crypts within the middle region of explants (5.2%) was considerably lower than that observed in vivo (8.8%). Crypt length and the distribution of labelled cells appeared similar for both situations. A computer simulation program for crypt-cell proliferation was devised, facilitating the modulation of a number of parameters including the cell-cycle time (Tc) and its component phases, the cut off position, and cell loss at mitosis. This simulation was employed to model continuous labelling (72 h) data obtained in vitro and provided an estimate of various kinetic parameters. Data for the middle region of explants was fitted with a Tc of 62 h, an S phase of 8 h and a cell loss factor (20%) which was consistent with histological findings. A fit to the experimental data obtained in vitro could be achieved by a model based upon a mode of cellular organisation known to occur within crypts in vivo. Therefore in vitro, the dynamic processes of crypt-cell proliferation and migration appear to be organised in the same manner as seen in vivo. PMID- 2567089 TI - Are all lymphoid blasts in the cell cycle? DNA synthesis in the lymphoid tissues of the dog. AB - Labelling index after one or repeated intravenous injections of 3H-thymidine was measured for various subpopulations of lymphatic cells in different canine lymphoid compartments and correlated with cell morphology. High doses of tritiated thymidine were injected and exposure times of up to 211 days were used. The labelling indices of lymphoid blasts were comparable in all tissues investigated. Labelling index varied from 100% in immunoblasts to 4% in small sized lymphocytes. Approximately 80% of immunoblasts were labelled 1 h after 3H thymidine application and 100% labelling was obtained after 12 h repetitive 3H thymidine labelling. In contrast with medium-sized and large lymphocytes, immunoblasts seem to be rapidly proliferating cells in the dog with almost no Go cells. PMID- 2567090 TI - Freeze-fracture analysis of endothelial cell membranes in rabbit carotid arteries subjected to short-term atherogenic stimuli. AB - Endothelial cell membranes of rabbit carotid arteries were examined by the freeze fracture technique. In the normal endothelium the mean densities of membrane bound vesicles were 75 vesicles/microns 2 on the luminal cell membrane and 102 vesicles/microns 2 on the abluminal membrane. Whilst the vesical openings on the luminal membrane were randomly distributed those on the abluminal membrane were typically ordered in a macular pattern with lines free of vesicles. Tight and gap junctions between endothelial cells were numerous. After stimulating the carotid arteries with weak electrical impulses, a technique used to induce enhanced endothelial permeability and the formation of atheromatous plaques after repeated stimulations (Betz et al. 1985), vesicle openings were reduced to 78 vesicles/microns 2 on abluminal membranes. Membranes on the luminal side and intercellular tight and gap junctions remained unchanged. PMID- 2567091 TI - Blood donors with indeterminate anti-p24gag reactivity in HIV-1 western blot: absence of infectivity to transfused patients and in virus culture. AB - During a follow-up period of 23-40 months, 7 regular blood donors had persistently, and 4 had intermittently indeterminate anti-p24gag reactivity in human immunodeficiency virus (HIV)-1 Western Blot. Serological testing and viral cultures revealed that these donors had no signs of infection for HIV-1, HIV-2, human T-cell lymphotropic virus (HTLV)-4, and HTLV-1. Extensive interviewing and physical examination of these donors revealed neither risk factors, nor signs of HIV infection in the tested donors. Ten recipients, who were transfused with blood products from 6 of these 11 anti-p24gag-positive donors, were traced back. Six months after transfusion, no serological or clinical signs of HIV-1, HIV-2, or HTLV-1 infection were observed in these patients. It is concluded that blood donors with persistent or intermittent anti-p24gag reactivity in HIV-1 Western Blot, without development of antibodies to other HIV-encoded proteins in later blood samples, do not transmit the described retroviruses to transfused patients. PMID- 2567092 TI - Glomerulonephritis and sinusitis. Hypocomplementemic glomerulonephritis associated with coagulase-negative staphylococcal pansinusitis and subdural empyema. AB - A 16-year-old male developed acute hypocomplementemic glomerulonephritis in association with pansinusitis and subdural empyema. Nephrotic-range proteinuria, serum complement, and renal function rapidly returned to normal with antimicrobial therapy and surgical drainage of both infected cavities. Culture of the exudate obtained during drainage of the frontal sinus yielded pure growth of coagulase-negative staphylococcus. This case documents the association of acute hypocomplementemic glomerulonephritis and pansinusitis-subdural empyema due to coagulase-negative staphylococcus, not previously described. PMID- 2567093 TI - [The treatment of allergic diseases]. AB - In allergies, causal and symptomatic treatments are possible. Among the causal measures, antigen elimination has to be mentioned on the first hand. If this is not possible (in pollinosis, in allergy to the house dust mite), hyposensitization is indicated. Effectiveness and undesirable actions of hyposensitization are discussed in detail. Glucocorticoids maybe used for the symptomatic relief of allergic symptoms. However, cautious use is advisable. In type I-allergies, histamine is the most important mediator. Therefore, inhibitors of mast cell degranulation and H1-blocking agents are the drugs of first choice. Non sedative acting compounds (astemizole, oxatomide, terfenadine) are preferred. PMID- 2567094 TI - The changing role of physician assistants. PMID- 2567095 TI - Temazepam and diphenhydramine in nursing homes. PMID- 2567096 TI - [Gastrointestinal hormones and cancers of the gastrointestinal tract]. PMID- 2567097 TI - Takayasus' arteritis. PMID- 2567098 TI - [Enzymologic diagnosis of cerebral lesions in generalized forms of meningococcal infection in the acute period]. AB - Cerebrospinal fluid was tested for several enzymes (CPK, AST, GGTP, BP, BG, AK) in 97 patients with generalized meningococcal infection with prevalent CNS affliction. Marked changes were detected even in the absence of clinical signs of cerebral lesion that was typical only of meningococcemia. Nevertheless, most pronounced changes were observed in a group of patients with cerebral edema and swelling. The changes in cerebrospinal fluid enzyme levels coincided with a reduced glucose and increased protein contents. In these cases, CSF hypertension increased CSF pressure to maximal levels. Most informative indices were singled out and their levels characteristic of acute phase of the disease established. PMID- 2567099 TI - [Morphofunctional changes of the neurosecretory system and microcirculatory bed of the hypothalamus in patients with chronic alcoholic intoxication of varying degree]. AB - Microcirculation was studied in the hypothalamus in acute alcohol intoxication. Blood flow was sharply decelerated with capillaries paretically dilated, overfilled with blood. Stasis, sludge formation and venue microthrombosis were directly related to the alcohol concentration in blood and urine. In neurosecretion areas the hemorrhagic sites were detected with neurosecretory cells activity sharply reduced. Chronical alcohol administration increased the number of functionally inactivated capillaries. These with luminal vesiculation were also found as were lymphoid cell infiltrates in venule walls. The number of capillaries inactivated in the neurosecretory system paralleled the number of dark cells that reflected the inhibition effects. PMID- 2567100 TI - Neurochemical transmission in the dorsal column nuclei. AB - The transmitter chemistry of the dorsal column nuclei is reviewed, with special emphasis on the monosynaptic component of the dorsal column-medial lemniscal pathway. It is maintained that in this anatomically addressed system concerned mainly with fast, secure sensory transmission, amino acids represent the predominant mechanism used for chemical relay of primary afferent impulses. The major excitatory primary afferent transmitter is most likely glutamic acid, whereas gamma-aminobutyric acid (GABA) fulfills adequately the role of transmitter of recurrent, postsynaptic and presynaptic inhibition. Recent immunohistochemical and physiological evidence indicates that 5 hydroxytryptamine, originating mainly from neurons of the raphe nuclei, plays a modulatory role in dorsal column transmission of innocuous sensory information. The basic synaptic elements involved in transmission across this relay, along with their corresponding chemical identities, are presented in the form of a speculative model. PMID- 2567101 TI - Suppression of prolactin release and mRNA accumulation by two novel dopamine agonist agents. AB - Two novel dopamine agonist drugs, CV 205-502 and CQP 201-403, have been investigated to compare their effects on prolactin secretion and prolactin mRNA accumulation in cultured rat pituitary cells. Both drugs gave dose-dependent suppression of prolactin release over a 24 h incubation period: when each drug was used at 100 nmol/l CV 205-502 and CQP 201-403 induced suppression to 8.9 +/- 1.7 and 10.2 +/- 1.8% of control release, respectively, compared to 26.7 +/- 4.8% of control with 100 nmol/l bromocriptine. There was no consistent effect on growth hormone release. Cytoplasmic accumulation of prolactin mRNA was also inhibited by both drugs at this concentration, to 50.2 +/- 5.5% of control values by CV 205-502 and to 67.4 +/- 8% of control by CQP 201-403, and to a similar extent by 100 nmol/l bromocriptine (50.6 +/- 9.1% of control). None of the drugs had any significant effect on GH mRNA levels. These data suggest that the agents exert their effect at a pretranslational stage of prolactin synthesis, as well as at the level of hormone release. PMID- 2567102 TI - Prospective multicentre study on the prediction of relapse after antithyroid drug treatment in patients with Graves' disease. AB - Graves' disease is an autoimmune disease characterized by a course of remission and relapse. Since the introduction of antithyroid drug treatment, various parameters have been tested for their ability to predict the clinical course of a patient with Graves' disease after drug withdrawal. Nearly all these studies were retrospective [corrected] and often yielded conflicting results. In a prospective multicentre study with a total of 451 patients, we investigated the significance of a variety of routine laboratory and clinical parameters for predicting a patient's clinical course. Patients who had positive TSH receptor antibodies activity at the end of therapy showed a significantly higher relapse rate than those without (P less than 0.001). However, the individual clinical course cannot be predicted exactly (sensitivity 0.49, specificity 0.73, N = 391). The measurement of microsomal (P = 0.99, sensitivity 0.37, specificity 0.63, N = 275) or thyroglobulin antibodies (P = 0.76, sensitivity 0.18, specificity 0.84, N = 304) at the end of antithyroid drug therapy did not show a statistically significant difference in the antibody titre between the patients of the relapse and those of the remission group. Additionally, HLA-DR typing (HLA-DR3: P = 0.37, sensitivity 0.36, specificity 0.58, N = 253) was proven to be unsuitable for predicting a patient's clinical course. Patients with abnormal suppression or an abnormal TRH test at the end of antithyroid drug therapy relapse significantly more often (P less than 0.001) than patients with normal suppression or normal TRH test. Patients with a large goitre also have a significantly (P less than 0.001) higher relapse rate than those with only a small enlargement. The sensitivity and specificity values of all these parameters, however, were too low to be useful for daily clinical decisions in the treatment of an individual patient. This is also true for the combinations of different parameters. Though the highest sensitivity value (0.94) was found for a combination of the suppression and the TRH test at the end of therapy, the very low specificity value (0.13) for this combination reduced its clinical usefulness. PMID- 2567103 TI - Anti-TSH antibodies in Graves' disease and their failure to interact with TSH receptor antibodies. AB - The occasional occurrence in sera of patients with Graves' disease of negative values in the assay for TSH receptor antibodies led to the discovery of endogenous antibodies to TSH. We examined the sera of approximately 2500 patients with Graves' disease. Eight positive sera were found. The IgG in all 8 sera showed higher binding with both bTSH and porcine TSH (pTSH) than with human TSH (hTSH). This means that autoantibodies to TSH in sera from patients with Graves' disease are rare and often directed towards heterologous bovine and porcine TSH. When hTSH levels were determined in sera of hyperthyroid patients with positive antibodies to hTSH, discrepancies in serum hTSH levels were observed when using different assay methods, i.e. hTSH levels were higher with the double-antibody technique, and lower with immunoradiometric assays. Antibodies in these sera showed higher binding to pTSH-alpha subunit than to -beta subunit. The binding of the two pTSH subunits with antibodies could be displaced by intact bTSH. Neither stimulation in Graves' disease nor blocking in primary hypothyroidism of TSH receptor antibodies interfered with the binding of the anti-TSH antibodies to 125I-labelled pTSH, pTSH-alpha, and pTSH-beta. Consequently, using this type of autoantibodies to TSH we were unable to obtain evidence that the TSH receptor antibodies of patients with Graves' disease was an anti-idiotype antibody against anti-TSH antibodies. PMID- 2567104 TI - Train-of-four fade during onset of neuromuscular block with nondepolarising neuromuscular blocking agents. AB - Fade in the train-of-four (TOF) responses during onset of neuromuscular block was studied following administration of atracurium (225 or 450 micrograms/kg), vecuronium (40 or 80 micrograms/kg), pancuronium (60 or 120 micrograms/kg) and tubocurarine (450 micrograms/kg). TOF ratios were measured at approximate heights of T1 (first response in the TOF) of 75, 50 and 25%. Fade in TOF increased as the height of T1 decreased, with maximum fade being observed at T1 of 25%. The greatest difference between relaxants was observed at T1 of 25%, vecuronium showing the least fade and pancuronium, atracurium and tubocurarine showing increasing fade, in that order. The difference between atracurium and tubocurarine or between vecuronium and pancuronium was not significant, but the degree of TOF fade was significantly greater with atracurium and tubocurarine in comparison to vecuronium or pancuronium. PMID- 2567106 TI - Nine "Conferences of the three institutes" (1958-1988). PMID- 2567105 TI - Haemodynamic responses during general anaesthesia for renal transplantation in patients with and without hypertensive disease. AB - Haemodynamic changes in 81 patients undergoing surgery for renal transplantation were studied. They were allocated to three groups depending on whether or not they had chronic hypertension and which drugs were used to control it. The patients in Group I (N = 18, 22%) were normotensive and were not receiving antihypertensive therapy, those in Group II (N = 21, 26%) were taking beta blockers and those in Group III (N = 42, 52%) both beta-blockers and vasodilating agents. Antihypertensive medication was continued as prescribed until surgery. No anticholinergic premedication was given. All patients received a standardized anaesthesia which included thiopentone, fentanyl, vecuronium and isoflurane. Mean arterial blood pressure and mean heart rate were lowest in Group I compared with the other groups immediately before induction, following vecuronium and thiopentone administration, and after tracheal intubation (P less than 0.05). After the 10-min induction period, blood pressure and heart rate values did not differ between the groups. Although before and during surgery and anaesthesia central venous pressure did not differ between the groups, CVP was higher in Group I postoperatively compared with the other groups (P less than 0.05). No serious anaesthesia-related complications occurred. PMID- 2567107 TI - Serotonin and peptide immunoneuromodulators: recent discoveries and new ideas. PMID- 2567108 TI - Regulation of Escherichia coli glutamine synthetase. PMID- 2567109 TI - Genetics of growth hormone and its disorders. PMID- 2567110 TI - Evaluation of coronary artery disease in the patient unable to exercise: alternatives to exercise stress testing. AB - Exercise stress testing is a well-established method for the diagnostic, prognostic, and functional assessment of patients with known or suspected CAD. A variety of alternative tests have been described in patients unable to perform leg exercise. Atrial pacing and dipyridamole imaging have been evaluated most extensively, and results compare favorably with those of exercise testing for diagnosing the presence of CAD. Both tests may be used to assess prognosis after myocardial infarction, and dipyridamole imaging may be useful in patients undergoing preoperative evaluation. The use of the cold pressor test and isometric handgrip exercise have also been described. However, the value of both tests is limited by a relatively low sensitivity for detecting the presence of CAD. Other testing modalities--arm ergometry, intravenous infusion of beta adrenergic agonists, and transthoracic pacing--show promise but require further assessment to confirm their value. PMID- 2567112 TI - Preclinical pharmacologic properties of dilevalol, an antihypertensive agent possessing selective beta 2 agonist-mediated vasodilation and beta antagonism. AB - Dilevalol is a vasodilator with selective partial beta 2 agonism and nonselective beta-antagonist activity. In contrast to its potent beta 2-agonist activity, dilevalol is essentially devoid of beta 1-agonist action, and is therefore distinct from pindolol and celiprolol. Dilevalol inhibits beta 1- and beta 2 adrenergic receptors nonselectively in dogs, rats and in isolated tissues, including human myocardium, and its beta-antagonist potency is similar to that of propranolol. In contrast to its beta-antagonist activity, it exerts minimal if any alpha-adrenergic blockade. Dilevalol is 300- to 1,000-fold more potent at beta- than at alpha 1-adrenergic receptors, including those on human myocardium and mammary arteries. At antihypertensive and vasodilator doses in rats and dogs, dilevalol does not inhibit alpha 1-adrenergic receptors. Oral doses of 2.5 to 50 mg/kg of dilevalol reduce blood pressure in spontaneously hypertensive rats. Heart rate is not significantly affected. Tolerance does not develop to the antihypertensive response. In contrast, beta blockers such as propranolol do not reduce blood pressure in this model. Antihypertensive activity is also observed in dogs with renal hypertension. The hemodynamic profile of dilevalol is consistent with its vasodilator properties. In spontaneously hypertensive rats, antihypertensive doses of dilevalol reduce peripheral resistance without affecting cardiac output. Dilevalol also improves the compliance and distensibility of the large arteries. The vasodilator and antihypertensive responses to dilevalol are mediated by a partial agonist action at vascular beta 2 receptors, because they are inhibited by the nonselective beta blocker propranolol as well as the beta 2-selective antagonist ICI 118,551. PMID- 2567111 TI - Effects of dilevalol on forearm circulation in essential hypertension. AB - In 6 patients with untreated hypertension of mild or moderate degree, dilevalol was infused in the brachial artery. Doses were calculated to produce plasma levels approximating those achieved after oral dosing (0.03, 0.07, 0.1, 0.3 micrograms.kg-1.min-1) and also to produce plasma levels exceeding oral dosing (0.5, 1.0, 2.0 micrograms.kg-1.min-1) without causing any blood pressure or heart rate changes. The effects of the infusion on forearm blood flow were assessed by venous occlusion plethysmography. Dilevalol caused a progressive increase in flow in all subjects up to the intermediate dose given, with the effect being attenuated when the dose was increased further. Simultaneous propranolol infusion reduced the increase in flow induced by dilevalol, shifting the dilevalol-induced vasodilation dose response curve to the right. Isoproterenol infusion caused a marked, dose-related increase in flow that was equally well reduced by simultaneous infusion of dilevalol or propranolol. These results indicate that dilevalol effectively blocks peripheral vascular beta receptors in humans. The drug also acts as a partial beta 2-receptor agonist, causing vasodilation which can be reduced by co-administration of propranolol. PMID- 2567113 TI - Influence of beta 2 agonism and beta 1 and beta 2 antagonism on adverse effects and plasma lipoproteins: results of a multicenter comparison of dilevalol and metoprolol. AB - Dilevalol combines vasodilation due to selective beta 2 agonism and nonselective beta antagonism. We studied 311 patients randomized to dilevalol and 138 to metoprolol in a multicenter trial. After a 4-week placebo washout, dilevalol was titrated from 200 to 1,600 mg once daily and metoprolol from 100 to 400 mg to a goal supine diastolic blood pressure less than 90 and greater than or equal to 10 mm Hg decrease from baseline. Responders were followed for 1 year. The average age of patients was 51 years; 72% were men and 54% were white. Both drugs reduced blood pressure effectively to a similar level. Fewer patients discontinued dilevalol than did those taking metoprolol (9 vs 16%; p less than 0.03). More metoprolol-treated patients withdrew because of depression (6 vs less than 1%; p = 0.03) and impotence (5 vs less than 1%; p = 0.03). Lipoprotein levels before and after treatment were measured in 99 patients treated for 53.5 weeks with dilevalol (mean dose 438 mg). Dilevalol increased high-density lipoprotein (HDL) cholesterol by 2.5 mg/dl to 47.2 (p = 0.05), reduced low-density lipoprotein (LDL) cholesterol by 2.5 mg/dl, increased HDL/LDL by 0.03, and decreased total cholesterol/HDL cholesterol by 0.18. Triglycerides increased by 21 mg/dl (p = 0.06). In patients with an initial HDL cholesterol less than 35 mg/dl, dilevalol increased it by 9 mg/dl. In patients treated with metoprolol, the only significant change (p = 0.02) was a 41.9-mg/dl increase in triglyceride levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567115 TI - Ulcer healing: moderate or strong acid inhibition? PMID- 2567114 TI - Effects of dilevalol, metoprolol and atenolol on left ventricular mass and function in nonelderly and elderly hypertensive patients. AB - Using serial M-mode echocardiographic determinations of left ventricular (LV) mass and function, the effects of dilevalol, a selective beta 2 agonist with nonselective beta-antagonist properties, were compared with those of metoprolol in 2 centers in double-blind, randomized clinical trials using similar protocols in nonelderly hypertensive patients (aged less than 65 years) (study 1). In a separate bicenter study with a similar design, dilevalol was compared with atenolol in elderly hypertensive patients (aged greater than or equal to 65 years) (study 2). Patients in both studies received placebo for 2 to 4 weeks, and were then randomized to receive increasing doses of dilevalol (200, 400, 800, 1,600 mg) or metoprolol (100, 200, 300, 400 mg) in study 1, and dilevalol (100, 200, 400, 800 mg) or atenolol (50, 100 mg) in study 2, to achieve a supine diastolic blood pressure (BP) of less than 90 mm Hg. In both studies, LV function and mass (Penn convention) were determined by echocardiographic examinations performed before randomization and at the end of the active treatment phase. Dilevalol, metoprolol and atenolol significantly reduced BP compared with placebo. In the nonelderly patients, a modest reduction in LV mass was observed with dilevalol (p less than 0.03) but not with metoprolol. Indexes of LV function -as assessed by end-diastolic and end-systolic dimensions, LV ejection time and ejection fraction--were better preserved by dilevalol than by metoprolol. In the elderly, neither dilevalol nor atenolol affected LV mass; however, indexes of LV function were better preserved with dilevalol than with atenolol.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567116 TI - DNA marker haplotype association with pancreatic sufficiency in cystic fibrosis. AB - Patients with cystic fibrosis (CF) generally suffer from chronic obstructive lung disease, pancreatic insufficiency (PI), and a number of other exocrine malfunctions. Approximately 15% of CF patients are, however, pancreatic sufficient. To investigate whether the two clinical subgroups, PI and pancreatic sufficiency (PS), are caused by different CF mutant alleles, we have performed linkage disequilibrium and haplotype association analysis with three DNA markers that are tightly linked to the CF locus. The study showed that the allelic and haplotype distributions for these RFLPs are significantly different between the two groups. The data suggest that most of the CF-PI patients are probably descendants of a single mutational event at the CF locus and that the CF-PS patients resulted from multiple, different mutations. While final interpretation of these data awaits molecular cloning of the CF gene, the information on haplotype association in CF may be useful in genetic counseling and disease prognosis, in identifying the gene itself, and in defining the mutations. PMID- 2567118 TI - Identification and characterization of nine RFLPs at the adenosine deaminase (ADA) locus. AB - We have identified and/or characterized at least nine RFLPs at the adenosine deaminase (ADA) locus, detected by digestion of DNA with MspI, BanII, PstI, BalI, and PvuII. The RFLPs were distributed over approximately 15 kb of the gene, from IVS 2 to IVS 10. They exhibited Mendelian inheritance and were in Hardy-Weinberg equilibrium. For seven fully characterized RFLPs, the gene frequencies of the rare alleles in 90 chromosomes examined ranged from .33 to .04, the PIC from .34 to .07, and the heterozygosity from .09 to .58. In kindreds examined (58 independent chromosomes), a total of nine haplotypes could be defined on the basis of seven fully characterized RFLPs with a heterozygosity of .62 and PIC of .53. Because there was considerable linkage disequilibrium, only three haplotypes accounted for 90% of individuals. Similar heterozygosity and PIC values (.59 and .51, respectively) could be obtained on the basis of haplotypes defined by the two sites that were the most polymorphic and that were in the least degree of linkage disequilibrium. A strategy for use of the RFLPs in linkage studies is suggested. We have also examined DNA from 17 patients with complete genetic deficiency of ADA (resulting in severe combined immunodeficiency [ADA-SCID] and from 10 patients with partial ADA deficiency (deficient in erythrocytes, with varying levels of ADA in other cells and normal immune function). Although the RFLPs detected genetic compounds among both types of patients, there was, as expected, a decreased incidence of heterozygosity (ADA-SCIDs, .29; partial ADA deficients, .20). Two additional haplotypes not found in the normal population were identified in homozygous form in patients. This information should be useful in developing a rational approach to delineation of mutations at the ADA locus as well as in distinguishing recurrent mutations of independent origin from those derived from a common progenitor. PMID- 2567119 TI - Allergic reactions to amyl nitrite inhalation. PMID- 2567117 TI - Evidence for mutation by unequal sister chromatid exchange in the Duchenne muscular dystrophy gene. AB - We have studied three families each containing a male with Duchenne or Becker muscular dystrophy. Southern blot analysis using both genomic and cDNA probes revealed that an exon-containing segment of DNA within the gene is duplicated in the probands, their mothers, and, in two cases, their sisters. The grandpaternal origin of the duplication has been demonstrated in these families by RFLP and duplication analysis. The results suggest that unequal sister-chromatid exchange, which most likely occurred in the germ cell lineage of the proband's grandfather, is responsible for generating these duplications and that this type of intrachromosomal rearrangement, although rarely reported in humans, is not uncommon in the muscular dystrophy gene. PMID- 2567121 TI - A prospective analysis of 24 episodes of neuroleptic malignant syndrome. AB - The authors prospectively studied 24 consecutive cases of neuroleptic malignant syndrome occurring in 20 patients in a general hospital over a 6-year period. They present detailed data concerning the clinical setting in which neuroleptic malignant syndrome occurs, the associated clinical and laboratory features, the favorable outcome that can be achieved with vigorous supportive therapy, and the factors that might help predict whether patients with neuroleptic malignant syndrome will subsequently be able to tolerate the reintroduction of neuroleptic medication. PMID- 2567122 TI - Changing patterns of neuroleptic dosage over a decade. AB - A study of patterns of neuroleptic dosage for 206 schizophrenic inpatients showed significant differences over time and among three centers--a general hospital psychiatric unit, a community mental health center, and a state hospital. In 1982 patients' mean dose at discharge was higher than the peak mean daily dose in 1973, and high-potency neuroleptics were being used almost exclusively. The mean length of stay decreased from 49 days in 1973 to 34 days in 1982. The possible relationship, if any, between increasing dosage, decreased length of stay, and the switch from low-potency to high-potency neuroleptics remains undetermined. PMID- 2567123 TI - Quinine for neuroleptic-induced dystonia and muscle stiffness. PMID- 2567120 TI - Identification of a deoxyribonucleic acid allelic variant for beta 1-4 galactosyltransferase expression associated with male sperm binding/penetration infertility. AB - Studies on mouse sperm-egg binding and fertilization have been suggested to involve the interaction of sperm-associated beta 1-4 galactosyltransferase with egg zona pellucida glycoproteins. A population of human males, whose sperm demonstrated an inability to penetrate ovulated zona pellucida-free hamster eggs in vitro, were examined for the level of activity of beta 1-4 galactosyltransferase. The level of enzyme activity was found to be reduced in human sperm isolated from this group of individuals compared with a known hamster penetration-positive group. Analysis of the deoxyribonucleic acid from these individuals by Southern hybridization with a putative human complementary deoxyribonucleic acid clone to beta 1-4 galactosyltransferase identified a unique allele lacking 0.8 and 0.4 kb restriction fragments on digestion with the endonuclease Taq I. These results represent the first evidence to suggest that mutations could be associated with the human gene for galactosyltransferase. Our data help to clarify one of the possible molecular mechanisms responsible for sperm-egg binding/penetration interactions. PMID- 2567124 TI - Experimental vertical transmission of Japanese encephalitis virus by Culex tritaeniorhynchus and other mosquitoes. AB - Vertical transmission of Japanese encephalitis virus to the F1 adult stage was demonstrated in Culex tritaeniorhynchus, Cx. annulus, Cx. quinquefasciatus, and Armigeres subalbatus. Transmission to the F1 larval stage was demonstrated in Cx. pipiens, Aedes vexans, Ae. alcasidi, and A. flavus. In Cx. tritaeniorhynchus, vertical transmission rates (the percentage of parent females transmitting to progeny) varied (12-100%). Filial infection rates (the percentage of progeny infected) for a given mosquito virus combination were markedly affected by the interval of time between parental infection and oviposition, suggesting that vertical infection was not transovarial in nature but occurred at oviposition. Filial infection rates for Cx. tritaeniorhynchus also varied widely by family and, as measured in F1 larvae, rates in excess of 20% were observed in a family. Filial infection rates in Cx. tritaeniorhynchus F1 adults were about 4 times lower than those in larvae. Japanese encephalitis virus was sexually transmitted from male to female Cx. tritaeniorhynchus. PMID- 2567125 TI - [Decubitus ulcers of the pelvic area. Diagnosis and surgical therapy]. PMID- 2567126 TI - [The effect of pancuronium and norcuron on hemodynamics, coronary circulation and myocardial oxygen consumption in coronary surgery patients]. AB - The effects of the non-depolarizing muscle relaxants pancuronium (Pancuronium) and vecuronium (Norcuron) (0.1 mg/kg) on myocardial blood flow, myocardial oxygen consumption, myocardial lactate balance, cardiovascular dynamics and electrocardiogram were studied in two groups of eight patients undergoing coronary artery bypass surgery. After an introduction of anaesthesia with 0.015 0.02 mg/kg rohypnol, isoflurane (0.5 Vol%) and N2O/O2 neuromuscular blockade was induced with pancuronium or vecuronium combined with a single dose of 0.005 mg/kg fentanyl. Measurements and electrocardiogram were performed before anaesthesia, in steady state of anaesthesia and after relaxation with pancuronium or vecuronium, combined with fentanyl. The measurements consisted of heart rate (HR), cardiac index (CI), stroke volume index (SVI), mean arterial pressure (AP), total peripheral resistance (TPR), pulmonary arterial pressure (PAP), pulmonary capillary wedge pressure (PCWP), right atrium pressure (RAP), myocardial blood flow (MBF), coronary vascular resistance (CVR), myocardial oxygen consumption (MVO2), coronary ateriomixed venous oxygen content difference (AVDO2cor), myocardial lactate extraction (LE) and rate pressure product (RPP). In the vecuronium group heart rate (HR) decreased significantly greater (21%) than in the pancuronium group (9%). Therefore myocardial oxygen consumption (MVO2) and coronary blood flow (CBF) diminished more in the vecuronium (48% resp. 35%) than in the pancuronium group (31% resp. 18%). The higher metabolic demand in the pancuronium group induced a significantly lower coronary vascular resistance (CVR). All the other hemodynamic parameters did not differ significantly in both patients groups. In all the patients we could not observe ST-segment depressions or elevations in the ECG, increases of PCWP or myocardial lactate productions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567127 TI - [Postoperative, opiate-induced respiratory depression is not dependent on arousal]. AB - INTRODUCTION: Respiratory depression after intravenous anesthesia is supposed to be related to vigilance. This hypothesis could not yet be tested because of the lack of methods measuring continuously important parameters of respiration without altering the patient's vigilance. Pulse Oximetry offers this possibility. The following study was performed to investigate the effects of the Benzodiazepine Antagonist Flumazenil (Anexate) on parameters of vigilance and respiration after Midazolam/Fentanyl anesthesia. METHODS: 40 healthy patients aged 18-65 years who were to undergo arthroscopy were randomly assigned to Flumazenil (Group F) or no Flumazenil = Control (Group C). All patients received 7.5-10 mg Midazolam, 0.3-0.6 Fentanyl, 4-6 mg Vecuronium, were intubated and ventilated with 8 ml/kg BW VT x 12/min - N2O/O2 = 2:1. At the end of operation Group F received 0.3-0.5 mg Flumazenil. When adequate spontaneous ventilation was restored the patients were extubated and brought to a single bed room where they were monitored and observed without being disturbed, except at the arrival time (T1), 15 min (T2) and 30 min (T3) when blood pressure was measured and the pain score was asked. The following parameters were registered: Transcutaneous Oxygen Saturation (SAT) and Heart Rate (HR) continuously, Sedation (every minute) and Reactions to acoustic or verbal stimuli in the case of hypoxemia. The two groups were compared with respect to the number and severity of hypoxic events/15 min, the mean degree of sedation (6 point scale) and the number of adequate reactions to the acoustic alarm resp. instruction: "Take a deep breath!" STATISTICS: Wilcoxon Test, Chi-Square test (p less than or equal to 0.05) is significant). RESULTS: The groups were comparable with respect to their anthropometric data, dosages of Midazolam and Fentanyl, and perioperative blood pressures. Parameters of vigilance: Group F was less sedated than Group C (p = 0.052) and reacted better to the verbal instruction to take a deep breath in the case of hypoxia (p less than or equal to 0.05). Parameters of respiration: Hypoxic states occurred more frequently in group F (p = 0.098) and lasted longer. The severeness was significantly more pronounced in group F (p less than or equal to 0.05). There were no complications and the patients acceptance of the anesthetic procedure was high. CONCLUSIONS: The hypothesis that postoperative respiratory depression is related to the degree of vigilance cannot be accepted. In contrast there is a strong evidence that under special conditions patients can be in a relatively high degree of vigilance and do not breathe with subsequent severe hypoxemia. The possible u PMID- 2567128 TI - [Methadone--pharmacokinetics and pharmacodynamics of an opiate]. AB - Methadone is described in this review as a potent analgesic agent; in Germany it is seldom administered as a therapeutic agent. It has the following pharmacokinetic properties: high lipophilicity, large volume of distribution (initial and steady state), low clearance (hepatic capacity limited) with a long terminal elimination time, high bioavailability following oral administration, and a tendency to accumulate in the blood and tissues. The review also considers pharmacodynamic aspects of methadone, which in Germany is available only in the levo-rotatory form; all the effects are conditioned by this l-enantiomer. In other countries methadone is successfully used over the following range of indications: intra- and postoperative pain (intravenous and epidural administration), cancer pain, and non-malignant painful conditions. The concepts of minimal effective analgesic concentration in the blood plasma (MEAC) and concentration required for 50% relief on pain are discussed. The MEAC is lower for the l-isomer than for the racemate. Methadone alone is a good analgesic agent that has slight effects in the form of respiratory depression, to which partial tolerance builds up in the course of long-term use of the drug. The interaction between methadone with alcohol, benzodiazepines, and barbiturates can be dangerous. PMID- 2567129 TI - Phycoerythrin fluorescence-based assay for peroxy radicals: a screen for biologically relevant protective agents. AB - Under the conditions of this assay, antioxidants that react rapidly with peroxy free radicals (e.g., ascorbate, vitamin E analogs, urate), protect phycoerythrin completely from damage by such radicals generated by thermal decomposition of 2,2'-azobis(2-amidinopropane); other compounds provide partial concentration dependent protection. Change in phycoerythrin fluorescence emission with time provides a measure of the rate of free radical damage. The assay exploits the unusual reactivity of phycoerythrin toward these peroxy radicals. On a molar basis, phycoerythrin reacts with these radicals over 100-fold slower than do ascorbate or vitamin E analogs, but over 60-fold faster than other proteins. Applications of this assay to the estimation of the peroxy radical scavenging capacity of human plasma are described, and to the comparison of the scavenging properties of several proteins and of DNA, of vitamins and their derivatives, of catecholamine neurotransmitters, and of a variety of other low molecular weight biological compounds. PMID- 2567130 TI - Determination of carbonyl groups in oxidatively modified proteins by reduction with tritiated sodium borohydride. AB - Oxidatively modified proteins have been implicated in a variety of physiologic and pathologic processes. Oxidative modification typically causes inactivation of enzymes and also the introduction of carbonyl groups into amino acid side chains of the protein. We describe a method to quantify oxidatively modified proteins through reduction of these carbonyl groups with tritiated borohydride. The technique was applied to purified, oxidatively modified glutamine synthetase and to bronchoalveolar lavage fluid from dogs and from humans. Since the protein content of lung lavage fluid is low, a very sensitive method was required to measure the oxidized residues. Reduction of the carbonyl group generated during oxidation of proteins with tritiated borohydride provided excellent sensitivity. Incorporation of tritium was directly proportional to the amount of protein with a range from 10 to 1000 micrograms. Should moieties other than amino acids be labeled, they are easily removed by rapid benchtop hydrolysis of the protein followed by chromatography on Dowex 50. PMID- 2567131 TI - A superfusion system designed to measure release of radiolabeled neurotransmitters on a subsecond time scale. AB - A new method for subsecond measurement of release of neurotransmitters from nerve terminal preparations (e.g., synaptosomes) in vitro is described. Synaptosomes were prelabeled with [3H]GABA via a Na-dependent GABA uptake system. The prelabeled nerve terminals are retained on small glass fiber filters in a superfusion chamber accessed by three high speed, solenoid-driven valves. Microcomputer-programmed circuitry controls the timing of valve operation. Each valve controls the delivery of a separate solution to the chamber, permitting rapid and independent control of membrane potential, [Ca2+]e, and drug delivery. The minimal dead volume of the chamber and the relatively high solution flow rate afford time resolution for release of at least 60 ms. This time resolution was necessary to observe the most rapid of at least three components of GABA release. PMID- 2567132 TI - Pain control during extracorporeal shock wave lithotripsy of gallstones by titrated alfentanil infusion. PMID- 2567133 TI - Modified rapid sequence induction. PMID- 2567134 TI - Delayed respiratory depression following alfentanil. PMID- 2567135 TI - The bovine gene map. AB - The present status of the bovine gene map as well as some of the methods and strategies important for future efforts in completing the gene map of cattle are reviewed. PMID- 2567136 TI - The analyses of antigen and DNA polymorphism within the bovine major histocompatibility complex: 1. The class I antigens. AB - Serology, isoelectric focusing (IEF) of expressed antigens, and restriction fragment length polymorphisms (RFLP) were compared for the identification of BoLA class I haplotypes. Expressed antigens identified as bands by IEF correlated well with serological definition confirming and extending our earlier findings (Joosten et al. 1988). Comparison of serology and isoelectric focusing bands with restriction fragments was more complicated; fragments were found which correlated both with broadly reacting and antigen specific sera. We also found correlation of fragments with two or more sera which showed no cross-reactivity. Fragments unique to particular haplotypes were also observed. Serology remains the simplest method of typing BoLA class I antigens. Isoelectric focusing generally agrees with serological definition of antigens and detects antigens not yet defined by serology. It may also be useful in defining the products of other expressed BoLA class I loci. In order to identify RFLPs which could be used for typing, comparison with serology or IEF is essential. Haplotype specific RFLPs could be useful in identifying genes linked to the MHC. PMID- 2567137 TI - Association of restriction fragment length polymorphisms of swine leucocyte antigen class I genes with production traits of Duroc and Hampshire boars. AB - Restriction fragment length polymorphism analyses of swine leucocyte antigen (SLA) class I genes were performed on 70 Duroc and 38 Hampshire boars from the 1986-87 national performance tests of each breed in the USA. Few boars were inbred. Southern blotting and hybridization procedures were performed on genomic DNA, isolated from white blood cells, using PvuII endonuclease and a swine major histocompatibility complex (MHC) class I probe. Durocs had an average of 11 restriction fragments, with the most common being in 63% of the boars and the least common appearing in only one boar. Hampshire boars had an average of 12 restriction fragments, with the most common appearing in 73% of the boars and the least common appearing in only one boar. Least squares procedures and stepwise regression methods were used to examine the association between DNA restriction fragments and the selection index (INDEX), average daily gain (ADG), average backfat thickness (BF), loin muscle area (LEA), and age at 104 kg (DAY104). In the Duroc breed one DNA restriction fragment was associated with decreased INDEX (P less than 0.05) and decreased ADG (P less than 0.05) whereas two other fragments were associated with increased BF (P less than 0.05). In the Hampshire breed two restriction fragments were associated with an increase in INDEX (P less than 0.05). Cluster analyses were used to group pigs of each breed on the basis of similar RFLP patterns. One cluster group in the Duroc breed was associated with lower average INDEX values (P less than 0.05), greater average DAY104 (P less than 0.05), and a larger mean LEA (P less than 0.05). In the Hampshire breed one cluster group was associated with lower INDEX (P less than 0.05). These results suggest there may be an association between swine MHC class I genes and performance traits in swine. The use of SLA class I restriction fragments, as genetic markers, may have potential in the future for improving pig performance. PMID- 2567138 TI - Alpha 2 antagonists. PMID- 2567139 TI - Virulence determinants of Salmonella typhimurium from animal sources. AB - Two hundred seventy-eight strains of Salmonella typhimurium isolated from 1973 to 1981 from animal sources in New York State were studied for possible virulence determinants and for a serotype-specific plasmid possibly linked with virulence. Of the strains, 98% possessed type-1 fimbriae. All strains possessed flagella and were motile. One hundred twenty-three strains (44%) treated with mitomycin C tested positive for the cholera-Escherichia coli heat labile family of toxins by a kinetics-based ELISA; when treated with mitomycin C and extracted with polymyxin B, 249 (90%) were positive in the kinetics-based ELISA. All strains were negative in the Biken test. A smooth cell wall was found in 99% of the strains. Sixty-one percent (169) of the strains had a 62-Md plasmid. Seventy-six (27%) of the strains had detectable plasmids ranging in size from 1 to 124 Md. PMID- 2567140 TI - [Determination of growth hormone: comparison between a physiologic stimulus (exercise) and pharmacologic stimuli (clonidine/guanfacine)]. AB - We have compared the release of growth hormone (GH) after exercise (10 minutes pedaling on a fixed bicycle with ergometer), and two pharmacological alpha adrenergic stimuli: clonidine (CL) or guanfacine (GU); in 36 prepuberal children whose height was below the third percentile. The tests were carried out in basal conditions, randomized and with a minimum interval of seven days between them. Blood GH was determined from cannulated antecubital veins. The results show a positive response (GH greater than 7 ng/ml) in 65.7% of cases after exercise, in 74.2% after CL and 71% after GU. The significant secondary effects observed after CL administration were sleepiness and decrease of arterial pressure. It is concluded that exercise is a useful test for assessing a possible deficiency of GH, and lacks significant side effects and that the diagnostic value of GU is similar to that of CL. PMID- 2567141 TI - Surreptitious ipecac administration simulating intestinal pseudo-obstruction. PMID- 2567142 TI - Predictive markers for the acquired immunodeficiency syndrome (AIDS) in hemophiliacs: persistence of p24 antigen and low T4 cell count. AB - STUDY OBJECTIVE: To investigate the predictive value of assays for human immunodeficiency virus (HIV) p24 antigen, p24 antibody, and gp120 antibody compared with T4 cell counts. DESIGN: Prospective cohort selected from persons who had HIV-antibody seroconversion. PATIENTS: Eighty-seven persons with hemophilia with an actuarial cumulative acquired immunodeficiency syndrome (AIDS) incidence of 26% (CI, 12% to 40%), 8 years after HIV-antibody seroconversion. INTERVENTION: None. MEASUREMENTS AND MAIN RESULTS: Human immunodeficiency virus p24 antigen was detected in 8 of 74 (11%) of the patients without AIDS and 7 of 13 (54%) of the patients with AIDS. The 2-year actuarial incidence of AIDS was 24% (CI, 0% to 48%) after detection of p24 antigen, 16% (CI, 0% to 34%) after loss of p24 antibody, 20% (CI, 0% to 45%) after loss of gp120 antibody, 31% (CI, 15% to 47%) after a T4 count of less than 200 cells/microL, and 67% (CI, 31% to 100%) after a T4 count of less than 200 cells/microL among those patients positive for p24 antigen. Very low numbers of T4 and T8 lymphocytes, presence of p24 antigen in serum, and absence of p24 antibody all had some predictive value. However, only p24 antigen (relative hazard 6.0, P = 0.008) and T4 counts (relative hazard 5.3, P = 0.002 with T4 count less than 200 cells/microL) independently predicted AIDS up to 12 months before diagnosis. CONCLUSIONS: Strong predictors of AIDS are p24 antigenemia or low T4 counts. Detection of p24 antigen is highly specific and complementary to the greater sensitivity of low T4 counts. These findings have important implications regarding prognosis, counseling, and the planning of clinical trials. PMID- 2567144 TI - Non-Q-wave myocardial infarction: pathophysiology, prognosis, and therapeutic strategy. AB - Non-Q-wave infarction, formerly referred to as nontransmural infarction, is usually associated with less myocardial necrosis and a lower in-hospital mortality than Q-wave or transmural infarction. Despite the initially favorable prognosis, however, long-term survival of patients after non-Q-wave infarction is similar to or even shorter than that after Q-wave infarction. It is now well documented that patients with non-Q-wave infarction are more prone to reinfarction, which usually occurs in the same area as the original injury. Since extensive damage from recurrent myocardial necrosis has a considerable deleterious effect on long-term survival, patients with non-Q-wave infarction would be expected to benefit from prophylactic treatment that prevents reinfarction. Indeed, this subset of patients appears to provide a unique opportunity to modify favorably the natural history of ischemic heart disease. PMID- 2567143 TI - Unique alterations of thyroid hormone indices in the acquired immunodeficiency syndrome (AIDS) AB - STUDY OBJECTIVE: To determine alterations in serum thyroid hormone indices in patients with human immunodeficiency virus (HIV) infection. DESIGN: Prospective, single-blind study. SETTING: Large metropolitan hospital where 20% of all patients with the acquired immunodeficiency syndrome (AIDS) in Los Angeles are treated. PATIENTS: Twenty-six inpatients with bronchoscopy-proven Pneumocystis carinii pneumonia and AIDS. Outpatients included 10 persons seropositive for HIV, 10 with AIDS-related complex, and 10 with AIDS. MAIN RESULTS: There were 19 survivors and 7 nonsurvivors of P. carinii infection. Serum triiodothyronine (T3) values generally remained normal until hospitalization, with nonsurvivors having lower values than survivors (0.56 +/- 0.1 nmol/L compared with 1.3 +/- 0.1 nmol/L, P less than 0.002, respectively). Reverse triiodothyronine (rT3) levels were low in persons with AIDS-related complex (0.21 +/- 0.02 nmol/L, P less than 0.001) and in AIDS outpatients (0.17 +/- 0.02 nmol/L, P less than 0.001). Normalization of rT3 occurred after patients were hospitalized (0.28 +/- 0.01 nmol/L). Serum thyroxine-binding globulin values rose with progression of HIV infection (seropositive, 369.7 +/- 18.1 nmol/L, P less than 0.005; AIDS-related complex, 419.1 +/- 37.0 nmol/L, P less than 0.005; AIDS, 423.3 +/- 31.9 nmol/L, P less than 0.005; survivors, 476.3 +/- 24.6 nmol/L, P less than 0.001), whereas nonsurvivors had normal values. All values are compared with normal values (T3, 2.3 +/- 0.04 nmol/L; rT3, 0.28 +/- 0.01 nmol/L; thyroxine-binding globulin, 288.2 +/- 6.9 nmol/L). CONCLUSIONS: Infection with HIV produces unique alterations in thyroid function. A progressive decline in rT3 and elevation in thyroxine-binding globulin accompany advancing HIV infection. The persistence of a normal T3 despite progression of HIV infection may contribute to weight loss. A low serum T3 on admission correlates with mortality. PMID- 2567145 TI - The neuropharmacology of attention-deficit hyperactivity disorder. AB - Attention-deficit hyperactivity disorder, as described in the American Psychiatric Association's diagnostic manual DSM III-R (1), is a broadly defined syndrome of cognitive and behavioral problems that were originally found in children, but that have been increasingly identified in adults who had similar complaints in childhood (2). This paper briefly reviews the definition and treatment of this syndrome in children, evidence for specific neurochemical abnormalities, and effects of stimulants on neurochemistry. Finally, problems of diagnosis and treatment in adults are discussed. PMID- 2567146 TI - Chemical coding of neurons and plurichemical transmission. PMID- 2567147 TI - Inactivation of two-electron reduced medium chain acyl-CoA dehydrogenase by 2 octynoyl-CoA. AB - The acetylenic thioester, 2-octynoyl-CoA, inactivates medium chain acyl-CoA dehydrogenase from pig kidney by two distinct pathways depending on the redox state of the FAD prosthetic group. Inactivation of the oxidized dehydrogenase occurs with labeling of an active site glutamate residue and elimination of CoASH. Incubation of the reduced dehydrogenase with 2-octynoyl-CoA rapidly forms a kinetically stable dihydroflavin species which is resistant to reoxidation using trans-2-octenoyl-CoA, molecular oxygen, or electron transferring flavoprotein. The reduced enzyme derivative shows extensive bleaching at 446 nm with shoulders at 320 and 380 nm. Denaturation of the reduced derivative in 80% methanol yields a mixture of products which was characterized by HPLC, by uv/vis, and by radiolabeling experiments. Approximately 20% of the flavin is recovered as oxidized FAD, about 40% is retained covalently attached to the protein, and the remainder is distributed between several species eluting after FAD on reverse phase HPLC. The spectrum of one of these species ressembles that of a N(5)-C(4a) dihydroflavin adduct. These data suggest that a primary reduced flavin species undergoes various rearrangements during release from the protein. The possibility that the inactive modified enzyme represents a covalent adduct between 2-octynoyl CoA and reduced flavin is discussed. Analogous experiments using enzyme substituted with 1,5-dihydro-5-deaza-FAD show rapid and quantitative reoxidation of the flavin by 0.5 eq of 2-octynoyl-CoA. PMID- 2567148 TI - [A molecular basis for multidrug resistance and reversal of the resistance]. AB - Multidrug-resistance is frequently characterized by enhanced drug efflux resulting from a membrane glycoprotein of 170,000 daltons (P-glycoprotein). Analysis of cloned cDNAs for the human MDR 1 gene, whose product is the P glycoprotein, indicates that P-glycoprotein is an energy-dependent drug-efflux system for cytotoxic hydrophobic anticancer drugs. We have demonstrated that a photoanalog of a reversing agent, SDB-ethylenediamine, specifically binds to P glycoprotein. The binding site on P-glycoprotein seems to be identical with that of anticancer agents and other reversing agents. On the other hand, the radioactive photoactive dihydropyridine calcium channel blocker, [3H] azidopine, photolabels P-glycoprotein in membrane vesicles from multidrug-resistant cells. This photolabeling is almost completely inhibited by excess dihydropyridine analogs that reverse or lower drug-resistance. In contrast, the labeling is not significantly inhibited by analogs that do not reverse resistance. These results suggest that it may be possible to quickly screen for dihydropyridine analogs that reverse multidrug resistance by measuring the inhibition of [3H] azidopine labeling of P-glycoprotein. PMID- 2567149 TI - Somatostatin analogue in short term management of hyperinsulinism. PMID- 2567150 TI - A double blind comparative study of sulphasalazine and hydroxychloroquine in rheumatoid arthritis: evidence of an earlier effect of sulphasalazine. AB - In a double blind, single observer, 48 week study the effects of sulphasalazine (2 g daily) and hydroxychloroquine (400 mg daily months 0-6, thereafter 200 mg daily) were compared in 60 patients with definite or classical rheumatoid arthritis. They had not been treated previously with second line drugs. The onset of response with sulphasalazine was earlier than with hydroxychloroquine. After 48 weeks a comparison of the treatments showed no statistically significant differences in disease activity variables. Adverse reaction was the main reason for withdrawal in the sulphasalazine group and lack of efficacy in the hydroxychloroquine group. All adverse reactions, one being agranulocytosis after eight weeks of sulphasalazine treatment, appeared in the first three months of treatment and were completely reversible. PMID- 2567151 TI - Alpha 1-adrenoceptor stimulation reduced the positive chronotropic effect of simultaneously administered beta-adrenoceptor- and H2-receptor agonists in guinea pig right atria. AB - The modulatory influence of alpha 1-adrenoceptor agonists on the beta adrenoceptor-mediated or H2-receptor-mediated positive chronotropic effect was investigated in isolated guinea-pig right atria. In the presence of cirazoline, the isoprenaline- or impromidine-induced positive chronotropic effect was significantly reduced. Methoxamine also reduced the isoprenaline-induced positive chronotropic effect. The reduction was abolished by the alpha 1-adrenoceptor antagonist prazosin. The protein kinase C activator phorbol-12-myristate-13 acetate, as well as the protein kinase inhibitor polymyxin B, had no influence on the cirazoline-evoked decrease of the beta-adrenoceptor-mediated positive chronotropic effect. Cirazoline, in the absence of isoprenaline or impromidine, concentration-dependently decreased heart rate, which could not be abolished by phentolamine and prazosin. Stimulation of alpha 1-adrenoceptors by phenylephrine in the presence of the beta-adrenoceptor antagonist sotalol, decreased spontaneous heart rate. The decrease was abolished by prazosin as well as by pretreatment with the neurotoxin 6-hydroxy-dopamine. The decrease in heart rate may to some extent be the result of interactions at the level of signal transduction systems of the 3 receptors. PMID- 2567152 TI - Highly selective and specific antagonism of central and peripheral alpha 2 adrenoceptors by atipamezole. AB - The potency, selectivity and specificity of atipamezole [MPV-1248, 4-(2-ethyl-2,3 dihydro-1H-inden-2-yl)-1H-imidazole], as an alpha 2-adrenoceptor antagonist was studied. In receptor binding studies [( 3H]-clonidine and [3H]-prazosin displacement) an alpha 2/alpha 1 selectivity ratio of 8526 was obtained for atipamezole, while idazoxan and yohimbine showed ratios of 27 and 40, respectively. Atipamezole had also about a 100 times higher affinity on alpha 2 adrenoceptors than the reference compounds. In the electrically stimulated prostatic portion of rat vas deferens, atipamezole showed potent competitive antagonistic activity against clonidine (pA2 8.6) and medetomidine (pA2 8.7) at presynaptic alpha 2-adrenoceptors. In the epididymal portion of the rat vas deferens, atipamezole had only weak competitive antagonistic activity against phenylephrine (pA2 5.0). In binding studies and studies with isolated organs, atipamezole had no effect on beta 1-, beta 2-, H1-, H2-, 5-HT1-, 5-HT2-, muscarine, DA2-, tryptamine, GABA, opiate and benzodiazepine receptors. In the pithed rat, atipamezole, like idazoxan, had partial alpha 1-adrenoceptor-mediated vasoconstrictor effects in addition to potent alpha 2-adrenoceptor blocking activity. In mice, medetomidine-induced sedation was effectively antagonized by atipamezole. These results show that atipamezole is a potent, selective and specific antagonist of both centrally and peripherally located alpha 2 adrenoceptors. PMID- 2567153 TI - Comparative study in mice of tetrazepam and other centrally active skeletal muscle relaxants. AB - Tetrazepam is a 1,4 benzodiazepine (BZD) clinically used in France and Germany as a muscle relaxant. The activity of tetrazepam was compared to that of diazepam, baclofen, mephenesin and chlormezanone in mice, in pharmacological models which are predictive of muscle relaxant and sedative properties. Tetrazepam was active in all the 6 tests of muscle relaxation (traction, chimney, inclined screen, grip force, horizontal grid and morphine-induced Straub tail). The overall muscle relaxant potency of tetrazepam was inferior to that of diazepam, but was clearly superior to that of chlormezanone and mephenesin. Baclofen was less active than tetrazepam in 3 tests (traction, horizontal grid, and grip strength), but more active in the other 3 tests. The administration of the benzodiazepine receptor antagonist Ro 15-1788 blocked the effects of tetrazepam and diazepam in 2 representative tests, morphine-induced Straub tail and the rotarod test, but did not modify the activities of the other centrally acting muscle relaxants in these same models. The selectivity ratio (ED50 rotarod or ED50 locomotor activity/ED50 in each muscle relaxant test) for tetrazepam was superior to that of diazepam and all the other muscle relaxant drugs examined. It is concluded that tetrazepam exerts its muscle relaxant activity by stimulating central BZD receptors, and presents the advantage of a wide dissociation between muscle relaxant and sedative potencies. PMID- 2567154 TI - Inadequate blockade by hexamethonium of the baroreceptor heart rate response in anesthetized and conscious rats. AB - The ability of hexamethonium (HEX) alone or in combination with atropine methylbromide (AMB) to block the heart rate (HR) response to baroreceptor activation by phenylephrine (PE) or angiotensin II (AII)-evoked increments in blood pressure was evaluated in rats. A highly significant negative correlation existed under control conditions between the changes in MAP and HR. HEX (10-20 mg/kg) did not influence the baroreceptor HR response of anesthetized and conscious rats, suggesting anesthesia was not involved in the failure of HEX to abolish the baroreceptor HR response. A combination of HEX and AMB (full ganglionic blockade) substantially attenuated but did not abolish the HR response to baroreceptor activation, but this effect was not significantly different from that produced by AMB alone. In a marked contrast to HEX, chlorisondamine (CHL) abolished the baroreceptor HR response both in anesthetized and in conscious rats. Efficacy of ganglionic blockade was tested by examining the effects of HEX or CHL on the frequency-bradycardic response relationship evoked by electrical stimulation of the peripheral end of the right vagus; HEX only attenuated the response whereas CHL abolished it. It is concluded that, even in doses which lower blood pressure, HEX: 1) does not adequately block the baroreceptor HR response; 2) when combined with a muscarinic blocker to induce full ganglionic blockade, the decrease in baroreceptor HR response is largely, if not totally, attributable to muscarinic blockade; 3) is much weaker than CHL in blocking ganglionic transmission and, even when combined with a muscarinic blocker, it does not abolish the baroreceptor HR response whereas CHL does, and 4) CHL represents a better choice for studies that require complete ganglionic blockade in anesthetized or conscious rats. PMID- 2567155 TI - Synthesis of 3-[2-(2,3-dihydro-5-phenyl-4-substituted-3H-1,2,4- triazole-3-thione 2-yl)-acetylamino]-2-methyl-4(3H)-quinazolinones and their pharmacological activities. AB - Some 2-methyl-3-triazole-substituted-4(3H)-quinazolinones 3a-f were prepared and tested for their H1- and H2-antihistaminic activities. In addition these compounds are central nervous system depressants and anticonvulsants. 3e shows highly significant decrease of locomotor activity. PMID- 2567156 TI - [Quinazolinones. 16. Synthesis and in vitro pharmacology of 2-aryl-1- ([3 (imidazol-4-yl)propyl]-guanidinylalkyl)-2,3-dihydro-4(1H)- quinazolinones]. AB - A series of 2-aryl-1- ([3-(imidazol-4-yl)propyl]guanidinylalkyl)-2,3-dihydro 4(1H)- quinazolinones were prepared starting with appropriate aminoalkyl quinazolinones. The substances proved to be moderate H1-antagonists at the isolated guinea-pig ileum as well as H2-agonists, maximally achieving 1.6 times the activity of histamine at the isolated guinea-pig right atrium. Compounds with a three-membered carbon chain connecting the bicyclus and the guanidine system are up to 100 times more active at the atrium (chronotropic effect) than the corresponding lower homologues. PMID- 2567157 TI - Transient primary hypothyroidism in the newborn: experience of the Victorian Neonatal Thyroid Screening Programme. AB - Between May 1977 and December 1986, the Victorian Thyroid Screening Programme tested approximately 570,000 newborns for congenital hypothyroidism. One hundred and sixty-six cases of primary hypothyroidism, confirmed by formal thyroid function tests, were identified, of which 24 were later found to be transient. In addition, there were two patients with permanent dyshormonogenesis who passed through a stage of being biochemically euthyroid and so could have been diagnosed mistakenly as transient hypothyroidism. Fourteen of the transient cases were due to excessive intake of iodine. In two, this was due to maternal ingestion of iodide during pregnancy and in 12 the babies received large amounts of topical iodine antiseptic. Two cases were caused by maternal anti-thyroid antibodies and in eight instances the cause was unknown. The large number of cases due to the topical application of iodine antiseptic emphasizes the need for caution when using this substance in neonates. PMID- 2567160 TI - Prevention of cardiac arrhythmias by adaptation to hypoxia: regulatory mechanisms and cardiotropic effect. AB - Adaptation to intermittent hypoxia had a pronounced antiarrhythmic effect in acute myocardial ischemia in conscious animals. This effect was less pronounced in anesthesia and was absent in isolated heart. In reperfusion, the prophylactic effect of adaptation was equally pronounced in all cases. Adaptation prevented stress-induced exhaustion of brain beta-endorphine presumably by its accumulation in adrenal glands and resulted in the accumulation of dopamine, 5 hydroxytryptamine and 5-hydroxyindolacetic acid in brain structures. These data naturally lead to the assumption that central mechanisms play the main role in the antiarrhythmic effect of adaptation to intermittent hypoxia on ischemic arrhythmias, while mechanisms occurring at the level of heart play the main role in the protective effect of the same adaptation against reperfusion arrhythmias. PMID- 2567159 TI - Participation of NMDA-receptors in ischemic changes of calcium homeostasis in rabbit brain. AB - The effects of nimodipine and of 2-amino-5-phosphonovalarate (APV) on Ca2+ fluxes in rabbit hippocampus during ischemia and during local application of N-methyl-D aspartate (NMDA) were studied by in vivo dialysis-perfusion technique. The drop of extracellular calcium evoked by ischemia was reduced from 30% in untreated animals to 7.5% by nimodipine applied i.v. and to 9% by local APV administration, whereas local nimodipine application had only slight effect. NMDA-evoked decrease of extracellular calcium concentration was highly sensitive to APV, whereas nimodipine had no effect when applied i.v. or only slightly reduced this effect during local administration. These results indicate that calcium channels coupled to NMDA-sensitive glutamate receptors, rather than voltage-dependent calcium channels play a key role in ischemic calcium influx to hippocampal neurons. PMID- 2567158 TI - Zonation of hepatic lipogenic enzymes identified by dual-digitonin-pulse perfusion. AB - The zonal distribution within rat liver of acetyl-CoA carboxylase, ATP citrate lyase and fatty acid synthase, the principal enzymes of fatty acid synthesis, was investigated by using dual-digitonin-pulse perfusion. Analysis of enzyme mass by immunoblotting revealed that, in normally feeding male rats, the periportal/perivenous ratio of acetyl-CoA carboxylase mass was 1.9. The periportal/perivenous ratio of ATP citrate-lyase mass was 1.4, and fatty acid synthase exhibited the largest periportal/perivenous mass gradient, having a ratio of 3.1. This pattern of enzyme distribution was observed in male rats only; in females, the periportal/perivenous ratio of enzyme mass was nearly equal. The periportal/perivenous gradients for acetyl-CoA carboxylase, ATP citrate-lyase and fatty acid synthase observed in fed (and fasted) males were abolished when animals were fasted (48 h) and refed (30 h) with a high-carbohydrate/low-fat diet. As determined by enzyme assay of eluates obtained from the livers of normally feeding male rats, there is also periportal zonation of acetyl-CoA carboxylase activity, expressed either as units per mg of eluted protein or units per mg of acetyl-CoA carboxylase protein, suggesting the existence of gradients in both enzyme mass and specific activity. From these results, we conclude that the enzymes of fatty acid synthesis are zonated periportally in the liver of the normally feeding male rat. PMID- 2567161 TI - Expression of rat renal gamma-glutamyltransferase cDNA in Escherichia coli. AB - To obtain the expression of rat kidney gamma-glutamyltransferase (GGT) cDNA in E. coli, plasmids containing the cDNA sequences coding for various parts of GGT were constructed. Transformation of E. coli cells by these hybrid vectors results in a production of unglycosylated recombinant proteins, immunologically recognized by specific antirat kidney GGT antibodies. Plasmid, expressing the complete coding sequence of GGT cDNA, allows the production of enzymatically active proteins localized in the periplasmic space, while the same sequence without the N terminal hydrophobic region results in a production of cytoplasmic proteins. These recombinant proteins present a very basic isoelectric point (pI greater than 9). These results suggest that the presence of the N-terminal region seems to be necessary to direct the expressed proteins enzymatically active in the periplasmic space. PMID- 2567162 TI - Thiyl radical attack on polyunsaturated fatty acids: a possible route to lipid peroxidation. AB - Absolute rate constants have been measured for the reaction of cysteinyl free radicals, CysS., with linoleic (18:2), linolenic (18:3) and arachidonic acid (20:4) in water/alcohol mixtures using the radiation chemical technique of pulse radiolysis. They are in the order of 10(6)-10(7) M-1 s-1 and increase with the number of biallylic functions, and with the polarity of the solvent. The reaction is shown to be a hydrogen atom abstraction from biallylic C-H bonds and yields pentadienyl radicals. The thiol mediated repair of the latter is considerably slower. Thiyl free radicals must consequently be considered as a potential source of lipid peroxidation. PMID- 2567163 TI - The metal requirement of rat tyrosine hydroxylase. AB - The effect of added metals on purified rat tyrosine hydroxylase which is predominantly iron-free has been determined. The presence of 10 microM ferrous ammonium sulfate results in a ten-fold increase in the activity of enzyme containing 0.1 iron atom per subunit. The enzyme activity is half-maximal at a free ferrous iron concentration of 0.15 microM. Copper, zinc, silver, and nickel are unable to replace ferrous iron. Ferric iron is inactive unless ascorbate is included to reduce it. PMID- 2567164 TI - Molecular cloning and amino acid sequence of rat kidney aminopeptidase M: a member of a super family of zinc-metallohydrolases. AB - Using a polyclonal antibody, a partial cDNA clone for rat aminopeptidase M was identified in a lambda gt11 library from rat kidney. A synthetic oligonucleotide probe derived from the sequence of the insert was used to screen a randomly primed lambda gt10 library. This allowed the identification of several overlapping clones encoding the full sequence of the enzyme. The reading frame, 2898 base pairs in length, encodes a 966 amino acid polypeptide. A highly hydrophobic segment, 24 amino acids in length, located close to the aminoterminus, is proposed to serve as the membrane-spanning domain for this membrane-bound enzyme. The sequence includes nine potential N-linked glycosylation sites and one potential sulfation site. In addition, the rat aminopeptidase M sequence contains an eight amino acid consensus sequence believed to serve as the zinc binding domain in a family of zinc metallohydrolases. Rat aminopeptidase M shows 77% similarity with the recently cloned human enzyme, as well as weaker but significant similarity with aminopeptidase N from E. coli (18%) and with human leukotriene A4 hydrolase (21%). PMID- 2567165 TI - Molecular basis of mouse Himalayan mutation. AB - Many different coat-colors result from the c-locus mutation in the mouse. One of these interesting mutants is a Himalayan, which produces temperature sensitive tyrosinase, and the basis of this sensitivity remains unknown. We cultured Himalayan mouse melanocytes from the skin and constructed a cDNA library; then, we isolated the Himalayan tyrosinase cDNAs and determined the nucleotide sequence. The tyrosinase gene in the Himalayan mouse contains an A----G change at nucleotide 1259 that alters a histidine residue to an arginine residue at amino acid 420. This histidine residue and the surrounding amino acids are conserved in their evolution from mouse to human. Interestingly, the residue with its surrounding eight amino acids are aligned between mouse b-protein and human tyrosinase. These results indicate the possibility that the altered residue at amino acid 420 of mouse tyrosinase may be important in stabilization of the tyrosinase molecule, or in interaction with other molecules, such as tyrosinase inhibitors. PMID- 2567166 TI - Loss of surface galactosyl receptor activity on isolated rat hepatocytes induced by monensin or chloroquine requires receptor internalization via a clathrin coated pit pathway. AB - We studied the effect of hyperosmotic inhibition of the clathrin coated pit cycle on the monensin- and chloroquine-dependent loss of surface galactosyl (Gal) receptor activity on isolated rat hepatocytes. Cells treated for 60 min without ligand at 37 degrees C with 25 microM monensin or 300 microM chloroquine in normal medium (osmolality congruent to 275 mmol/kg) bound 40-60% less 125I-asialo orosomucoid (ASOR) at 4 degrees C than untreated cells. Cells exposed to monensin or chloroquine retained progressively more surface Gal receptor activity, however, when the osmolality of the medium was increased above 400 mmol/kg (using sucrose as osmolite) 10 min prior to and during drug treatment. Cells pretreated for 10 min with hyperosmolal media (600 mmol/kg) alone internalized less than or equal to 10% of surface-bound 125I-ASOR. Thus, the ligand-independent loss of surface Gal receptor activity on monensin- and chloroquine-treated hepatocytes requires internalization of constitutively recycling receptors via a coated pit pathway. PMID- 2567167 TI - Under anaerobic conditions, soluble guanylate cyclase is specifically stimulated by glutathione. AB - Various thiols exert non-specific effects on the activity of soluble guanylate cyclase under aerobic conditions. We studied the effects of thiols under anaerobic conditions (pO2 less than 6 Torr) on soluble guanylate cyclase, purified from bovine lung. Reduced glutathione stimulated the enzyme concentration-dependently with half-maximal enzyme stimulation at a concentration of about 0.5 mM. The extend of maximal enzyme stimulation (up to 80-fold) was comparable with the activation by NO-containing substances. The activation by glutathione was additive with the effect of sodium nitroprusside. Cysteine and various other thiols increased the enzyme activity 20-fold and 2- to 5-fold, respectively. The stimulatory effect of these thiols was not related to their reducing potency. Activation of soluble guanylate cyclase by glutathione was dose dependently reduced in the presence of other thiols (cysteine greater than oxidized glutathione greater than S-methyl glutathione). Under aerobic conditions or with Mn-GTP as substrate, the effect of glutathione on soluble guanylate cyclase was suppressed. The results suggest a specific role for glutathione in the regulation of soluble guanylate cyclase activity and a modulation of this effect by redox reactions and other intracellular thiols. PMID- 2567168 TI - Structure-activity relationships of 1H-indole-7-carboxamides as a novel series of selective alpha 1-adrenoceptor agonists. AB - The present isolated tissue study was designed to quantitate the alpha adrenoceptor agonist activity of AY- 30,191 (5-(1-hydroxy-2-amino-ethyl)-1H indole-7-carboxamide) and a series of related compounds. AY-30,191 induced contractions in the rabbit aorta, which were blocked by prazosin. In the rat vas deferens, while clonidine inhibited the electrically induced twitch response, AY 30,191 caused a prazosin-sensitive augmentation. In the dog saphenous vein, rauwolscine was less effective than the combination of rauwolscine and prazosin in inhibiting the contractions induced by AY-30,191. Pretreatment of the dog saphenous vein with phenoxybenzamine reduced the response to AY-30,191. The addition of rauwolscine to phenoxybenzamine-treated tissues had no effect on the contractions to AY-30,191 remaining after phenoxybenzamine treatment. These results suggest that AY-30,191 is a selective alpha 1-adrenoceptor agonist. Optimal alpha 1-adrenoceptor agonist activity in the 1H-indole-7-carboxamide series was seen in compounds in which a) the indole ring and the ethylamine side chain were intact; b) the indole nitrogen was unsubstituted; and c) the carboxamide was present at the 7-position in the indole ring. Removal of the carboxamide decreased alpha 1-adrenoceptor activity and, more importantly, resulted in a loss of alpha 1-adrenoceptor selectivity. Replacement of the carboxamide in the 7 position with methanesulfonamide resulted in a decrease in activity but a retention of alpha 1-adrenoceptor selectivity, whereas the dimethylamino analog was nonselective and the phosphoramidic acid diethylester analog was inactive.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567169 TI - General pharmacology of nizatidine in animals. AB - Nizatidine (N-[2-[[[2-[(dimethylamino)methyl-4-thiazolyl]- methyl]thio]ethyl]-N' methyl-2-nitro-1,1-ethenediamine, LY139037, Axid) is a novel, potent, and selective H2-antagonist. The potential secondary pharmacologic effects of this agent on the cardiovascular, respiratory, gastrointestinal, renal, hepatic, autonomic, and central nervous systems as well as effects on circulating blood glucose and the acute inflammatory response were examined. Nizatidine was generally inactive in the tests conducted in mice, rats, guinea pigs, rabbits, and dogs. Nizatidine and the reference H2-antagonist, cimetidine, both produced effects upon the cardiovascular and respiratory systems by intravenous administration in anesthetized dogs at doses in excess of the intended clinical exposure. In summary, these studies confirm the selective pharmacologic activity of nizatidine and indicate a low potential for secondary pharmacologic side effects to be encountered clinically. PMID- 2567170 TI - A novel class of local antiinflammatory steroids. 2nd communication: pharmacological studies of methyl 11 beta,17 alpha,21- trihydroxy-3,20-dioxo pregna-1,4-diene-16 alpha-carboxylate and methyl 11 beta,21-dihydroxy-3,20-dioxo pregna-1,4-diene-16 alpha-carboxylate. AB - Two novel 16-substituted steroidal carboxylate esters derived from prednisolone, methyl 11 beta,17 alpha,21- trihydroxy-3,20-dioxo-pregna-1,4-diene-16 alpha carboxylate (P16CM) and methyl 11 beta,21-dihydroxy-3,20-dioxo-pregna-1,4-diene 16 alpha- carboxylate (DeoxyP16CM) were evaluated for in vivo antiinflammatory and glucocorticoid activities. Results indicate that incorporation of a methoxycarbonyl group at the 16 position of prednisolone, as in P16CM, resulted in 5.5 times more local activity in the cotton pellet granuloma assay and 14 times more topical activity in the croton oil induced ear edema bioassay as compared with the parent compound prednisolone (P). The 17 alpha-dehydroxy analogue of P16CM (DeoxyP16CM) retained one-half the local activity of P in the cotton pellet granuloma bioassay and topical activity equal to P in the croton oil induced ear edema bioassay. Favorable dissociation of local from systemic effects is seen for these steroidal 16-carboxylate esters since their systemic antiinflammatory activity was significantly less than that of P, and their suppression of plasma corticosterone and ACTH levels was minimal. While P16CM does exhibit some thymolytic activity, DeoxyP16CM is essentially devoid of thymus atrophogenic effects at equiactive doses. Thus, these compounds may represent safer topical therapeutic agents. PMID- 2567171 TI - Longitudinal study on pharmacodynamics and pharmacokinetics of acute, steady state and withdrawn quazepam. AB - To assess pharmacodynamic and pharmacokinetic properties of acute, subchronic and withdrawn quazepam, a single-blind, longitudinal study was run in eight male, healthy young volunteers. The design covered a 1-week placebo run-in period, a period with daily oral night-time administration of 15 mg quazepam until a pharmacokinetic steady-state was reached (3 weeks) and a 2-week placebo withdrawal period. Oculodynamic Test (ODT) (EOG-registration with simultaneous choice reaction task, CRT) and Adaptive Pursuit Tracking Test (APTT) were used for assessment of intradiurnal and long-term profiles of attention, perception, cognition, objective sedation, psychomotor and muscular (force-related) parameters and cardiorespiratory measures under workload. Visual analogue scales (VAS) of sedation, excitation and state anxiety were applied additionally. Plasma levels of quazepam and its metabolites (oxoquazepam and desalkyl-oxoquazepam) were intermittently analyzed by GC, within 24 h after actual blood sampling in the morning of assessment days, to check the attainment of the intended criterion for termination of medication (steady-state, "on-line kinetic procedure"). Adverse effects were recorded by subjects' written free recall and a symptom checklist. Although a pharmacokinetic steady-state could be reached in sequence for the parent drug quazepam and its metabolites within 3 weeks, there was no pharmacodynamic steady-state at the end of this period, but a continuous impairment in oculomotor variables. Performance in the choice reaction task and the APTT showed a similar tendency, which was masked to a certain extend by learning effects. There were no signs for rebound effects within the 2 weeks after withdrawal. Relevant carry-over phenomena declined after 3 days of withdrawal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567172 TI - Effective method for prediction of transient hypothyroidism in neonates born to mothers with chronic thyroiditis. AB - An effective method of prediction of neonatal transient hypothyroidism was examined in 105 neonates (including a pair of twins) born to mothers with chronic thyroiditis (92 mothers with goitrous Hashimoto's disease and 12 with primary atrophic hypothyroidism). Antithyroid microsomal antibody was measured by a hemagglutination technique (MCHA), and antithyroid-stimulating hormone (TSH) receptor antibody by both radioreceptor assay (TBII) and biologic thyroid stimulation blocking assay (TSBAb). For generalization of predictive criteria, the expression of TBII activity was standardized using standard serum made taking units of MRC-LATS-standard B as a reference, and that of TSBAb activity was standardized as the degree of dilution with normal pooled serum to attain 50% inhibition of TSH (100 microU/ml)-induced cyclic adenosine monophosphate increase (TSBAb50). The MCHA titer in maternal serum at delivery correlated well with that of the corresponding cord serum, but not with the free thyroxine (T4) index or the TSH level in cord serum. TBII activity was positive in only 4 of 12 mothers with primary atrophic hypothyroidism, TSBAb activity was also positive only in these four mothers, and neonatal thyroid dysfunction was observed in three of their neonates. Two of these neonates developed transient hypothyroidism requiring T4 treatment, and the t third developed mild transient hyperthyrotropinemia with normal T4 and triidothyronine levels. The mothers whose neonates showed transient hypothyroidism had TBII activities of more than 300 U/ml and TSBAb50 activities of more than 300. Ninety-two mothers with goitrous Hashimoto's disease had neither TBII nor TSBAb activity, irrespective of their thyroid function, and delivered euthyroid babies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567173 TI - Activation of the cellular Harvey ras gene in mouse skin tumors initiated with urethane. AB - Mouse skin tumors, benign papillomas, and squamous cell carcinomas (SCCs) were initiated by a single topical application of urethane followed by repeated promotion with 12-O-tetradecanoylphorbol-13-acetate (TPA). Using the NIH 3T3 focus forming assay, dominant transforming activity was detected in DNA isolated from SCC samples. Rearranged and amplified copies of the c-Ha-ras gene were detected in NIH 3T3 transformant cell lines, indicating that an activated Ha-ras gene had been transferred to the NIH 3T3 recipient cells. Analysis of p21ras from the transformant cell lines suggested that the activating ras mutation was present in codon 61. Ultimately, the Ha-ras gene was shown to be activated by a specific A----T transversion at the second position of codon 61. This mutation was detected in both benign papillomas and SCCs, suggesting the activation occurred early in tumor development. The results demonstrate a highly consistent activation of the Ha-ras oncogene by a specific point mutation, suggesting a functional role for an activated ras gene in the initiation of mouse skin tumors by urethane. PMID- 2567174 TI - Mechanism of spontaneous loss of heat-stable toxin (STa) production in enterotoxigenic Escherichia coli. AB - Five strains of enterotoxigenic Escherichia coli (ETEC) showing spontaneous loss of heat-stable enterotoxin (STa) production were studied to elucidate the underlying genetic mechanisms. Southern blot analysis revealed that loss of STa production, and the corresponding lack of hybridization with the STa gene probes, were associated with deletions of DNA fragments harboring the relevant toxin genes rather than with loss of plasmids. PMID- 2567175 TI - Antigenic targets in epimembranous glomerulonephritis. Experimental data and potential application in human pathology. AB - Although membranous glomerulonephritis (MGN) has been long considered as a prototype of glomerulonephritis (GN) due to the deposition of circulating immune complexes (CIC), a growing body of evidence indicates that immune deposits can also be formed in situ and implicate antigens expressed by glomerular epithelial cells (GEC). Some of these antigens have recently been identified. The first one, gp330 - a 330-kd glycoprotein restricted to the coated pits of GEC and renal brush border (BB) - is responsible for Heymann nephritis, a rat model of MGN. However, it is absent in the human glomerulus and is therefore probably not involved in human cases of MGN, at least in those due to in situ formation of CIC. In addition, by raising monoclonal antibodies against rat and rabbit BB, we have isolated two other BB proteins also expressed on GEC. The latter, respectively identified as dipeptidyl peptidase IV (90 kd) and enkephalinase (85 kd), can serve as targets for the formation of short-lived immune deposits. Since they are also detected on human GEC, they might play a role in the pathogenesis of MGN in man. PMID- 2567176 TI - Distribution of calcitonin- and somatostatin-containing cells in thyroid lymphoma and in Hashimoto's thyroiditis. AB - Eleven cases of thyroid lymphoma were studied by the immunoperoxidase avidin biotin technique with calcitonin and somatostatin rabbit antisera. In 6 cases of non-Hodgkin lymphoma, in thyroid tissue residual to the lymphomatous infiltration, the C cell density was markedly increased and clustering was often observed; the C cells often took part in the follicular lining, frequently with polar distribution; these elements displayed a strong positivity for calcitonin, while the number of somatostatin-containing cells was lower and the staining less intense. In the only case of Hodgkin's lymphoma of the thyroid gland the staining was negative; in other 4 cases of non-Hodgkin lymphoma no residual thyroid tissue was found and the staining was also negative. As Hashimoto's thyroiditis is often associated with thyroid lymphoma, 13 cases of Hashimoto's thyroiditis were also studied; no C cells were observed and both stainings were negative. These data show that an increase in the C cell number may be a hallmark of thyroid lymphoma and that hyperplastic C cells show an intensive positivity for calcitonin. On the other hand, C cell hyperplasia is not present in Hashimoto's thyroiditis, in spite of the close association with thyroid lymphoma. Furthermore, we provide evidence that somatostatin-containing cells are present both in normal thyroid glands and in thyroid lymphoma. PMID- 2567177 TI - Origin of the HIV-susceptible human CD4+ cell line H9. PMID- 2567178 TI - Antagonism of moderate degrees of vecuronium-induced neuromuscular block by small doses of neostigmine. AB - We have studied the influence of a reduced dose of neostigmine on recovery from vecuronium-induced neuromuscular block in 26 adult patients, using electromyographic responses to train-of-four (TOF) stimulation. Neostigmine 10, 20 or 40 micrograms kg-1 was administered when the first response had recovered spontaneously to 5-10% or 40-50% of control. Antagonism was accepted as adequate when the first response reached 90% of control and the TOF ratio reached 0.7. At both degrees of spontaneous recovery, neostigmine 40 micrograms kg-1 evoked the most rapid antagonism. Clinical recovery was satisfactory with no differences between groups. Block produced by neostigmine was not observed. The pattern of recovery of the single response and the TOF ratio was not altered by neostigmine in the range of doses studied. We suggest that the dose of neostigmine should not be reduced below 40 micrograms kg-1 even when all responses of the TOF are present. PMID- 2567179 TI - Ultrastructure of an arterial lesion induced in rats by fenoldopam mesylate, a dopaminergic vasodilator. AB - Fenoldopam mesylate (FM) is a dopaminergic vasodilator with demonstrated efficacy and a favourable safety profile in hypertensive and congestive heart failure patients. FM produced a novel arterial lesion in renal and splanchnic arteries of rats, but not dogs or monkeys. The studies reported here were undertaken to investigate the ultrastructure of the arterial lesion induced in rats by FM in an attempt to shed light on its pathogenesis. Rats were infused intravenously with FM, either 50 micrograms/kg/min for 1 or 4 h, or 5 or 100 micrograms/kg/min for 24 h. Control rats were infused for 4 or 24 h with vehicle alone. Perfusion-fixed tissue from the stomach and pancreas of control and drug-treated rats was examined by transmission electron microscopy. No arterial lesions were seen in rats infused with the drug for 1 or 4 h, or in control rats. All drug-treated rats infused with 5 or 100 micrograms/kg/min of FM for 24 h had lesions in subserosal gastric arteries and interlobular pancreatic arteries. In areas of mild arterial damage, medial smooth muscle cells contained intracytoplasmic pseudovacuoles, autophagic vacuoles, and electron-dense, myofilamentous inclusions. More severe lesions were characterized by overt medial necrosis and haemorrhage. The endothelium of affected arteries was invariably intact, except in areas of severe medial damage. The internal elastic lamina and connective tissue elements within the arterial wall were unaffected. These findings suggest that medial smooth muscle cells are the primary site of damage caused by fenoldopam mesylate in splanchnic arteries of the rat. This iatrogenic arterial lesion could provide an interesting model to study the response of medial smooth muscle to pharmacologically mediated injury. PMID- 2567180 TI - Calitoxin, a neurotoxic peptide from the sea anemone Calliactis parasitica: amino acid sequence and electrophysiological properties. AB - We have isolated a new toxin, calitoxin (CLX), from the sea anemone Calliactis parasitica whose amino acid sequence differs greatly from that of other sea anemone toxins. The polypeptide chain contains 46 amino acid residues, with a molecular mass of 4886 Da and an isoelectric point at pH 5.4. The amino acid sequence determined by Edman degradation of the reduced, S-carboxymethylated polypeptide chain and tryptic and chymotryptic peptides is Ile-Glu-Cys-Lys-Cys Glu-Gly-Asp-Ala-Pro-Asp-Leu-Ser-His-Met-Thr-Gly-Thr- Val-Tyr - Phe-Ser-Cys-Lys Gly-Gly-Asp-Gly-Ser-Trp-Ser-Lys-Cys-Asn-Thr-Tyr-Thr-Ala- Val-Ala - Asp-Cys-Cys His-Glu-Ala. No cysteine residues were present in the peptide. Similarly to other sea anemone toxins, calitoxin interacts, in crustacean nerve muscle preparations, with axonal and not with muscle membranes, inducing a massive release of neurotransmitter that causes a strong muscle contraction. The low homology of CLX with RP II and ATX II toxins has implications regarding the role played by particular amino acid residues. PMID- 2567181 TI - Characterization of site-directed mutants in the lac permease of Escherichia coli. 2. Glutamate-325 replacements. AB - lac permease with Ala in place of Glu325 was solubilized from the membrane, purified, and reconstituted into proteoliposomes. The reconstituted molecule is completely unable to catalyze lactose/H+ symport but catalyzes exchange and counterflow at least as well as wild-type permease. In addition, Ala325 permease catalyzes downhill lactose influx without concomitant H+ translocation and binds p-nitrophenyl alpha-D-galactopyranoside with a KD only slightly higher than that of wild-type permease. Studies with right-side-out membrane vesicles demonstrate that replacement of Glu325 with Gln, His, Val, Cys, or Trp results in behavior similar to that observed with Ala in place of Glu325. On the other hand, permease with Asp in place of Glu325 catalyzes lactose/H+ symport about 20% as well as wild-type permease. The results indicate that an acidic residue at position 325 is essential for lactose/H+ symport and that hydrogen bonding at this position is insufficient. Taken together with previous results and those presented in the following paper [Lee, J. A., Puttner, I. B., & Kaback, H. R. (1989) Biochemistry (third paper of three in this issue)], the findings are consistent with the idea that Arg302, His322, and Glu325 may be components of a H+ relay system that plays an important role in the coupled translocation of lactose and H+. PMID- 2567182 TI - Effect of distance and orientation between arginine-302, histidine-322, and glutamate-325 on the activity of lac permease from Escherichia coli. AB - lac permease of Escherichia coli was modified by site-directed mutagenesis in order to investigate the effects of polarity, distance, and orientation between the components of a putative H+ relay system (Arg302/His322/Glu325) postulated to be involved in lactose-coupled H+ translocation. The importance of polarity between His322 and Glu325 was studied by interchanging the residues, and the modified permease--H322E/E325H--is inactive in all modes of translocation. The effect of distance and/or orientation between His322 and Glu325 was investigated by interchanging Glu325 with Val326, thereby moving the carboxylate one residue around putative helix X. The resulting permease molecule--E325V/V326E--is also completely inactive; control mutations, E325V [Carrasco, N., Puttner, I. B., Antes, L. M., Lee, J. A., Larigan, J. D., Lolkema, J. S., Roepe, P. D., & Kaback, H. R. (1989) Biochemistry (second paper of three in this issue)], and E325A/V326E, indicate that a Glu residue at position 326 inactivates the permease. The wild-type orientation between His and Glu was then restored by further mutation of E325V/V326E to introduce a His residue into position 323 or by interchanging Met323 with His322. The resulting permease molecules- M323H/E325V/V326E and H322M/M323H/E325V/V326E--contain the wild-type His/Glu orientation, but the His/Glu ion pair is rotated about the helical axis by 100 degrees relative to Arg302 in putative helix IX. Both mutants are inactive with respect to all modes of translocation. The results provide strong support for the contention that the polarity between His322 and Glu325 and the geometric relationship between Arg302, His322, and Glu325 are critical for permease activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567183 TI - Solution structure of recombinant hirudin and the Lys-47----Glu mutant: a nuclear magnetic resonance and hybrid distance geometry-dynamical simulated annealing study. AB - The solution structure of recombinant wild-type hirudin and of the putative active site mutant Lys-47----Glu has been investigated by nuclear magnetic resonance (NMR) spectroscopy at 600 MHz. The 1H NMR spectra of the two hirudin variants are assigned in a sequential manner with a combination of two dimensional NMR techniques. Some assignments made in our previous paper [Sukumaran, D. K., Clore, G. M., Preuss, A., Zarbock, J., & Gronenborn, A. M. (1987) Biochemistry 26, 333-338] were found to be incorrect and are now corrected. Analysis of the NOE data indicates that hirudin consists of an N terminal compact domain (residues 1-49) held together by three disulfide linkages and a disordered C-terminal tail (residues 50-65) which does not fold back on the rest of the protein. This last observation corrects conclusions drawn by us previously on hirudin extracted from its natural source, the leech Hirudo medicinalis. The improved sensitivity of the 600-MHz spectrometer relative to that of our old 500-MHz spectrometer, the availability of two variants with slightly different chemical shifts, and the additional information arising from stereospecific assignments of methylene beta-protons and methyl protons of valine have permitted the determination of the solution structure of hirudin with much greater precision than before. Structure calculations on the N-terminal domain using the hybrid distance geometry-dynamical simulated annealing method were based on 685 and 661 approximate interproton distance restraints derived from nuclear Overhauser enhancement (NOE) data for the wild-type and mutant hirudin, respectively, together with 16 distance restraints for 8 backbone hydrogen bonds identified on the basis of NOE and amide NH exchange data and 26 phi backbone and 18 chi 1 side-chain torsion angle restraints derived from NOE and three-bond coupling constant data. A total of 32 structures were computed for both the wild type and mutant hirudin. The structure of residues 2-30 and 37-48 which form the core of the N-terminal domain is well determined in both cases with an average atomic rms difference between the individual structures and the respective mean structures of approximately 0.7 A for the backbone atoms and approximately 1 A for all atoms. As found previously, the orientation of the exposed finger of antiparallel beta-sheet (residues 31-36) with respect to the core could not be determined on the basis of the present data due to the absence of any long-range NOEs between the exposed finger and the core.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567184 TI - N-methyl-D-aspartate-sensitive [3H]glutamate binding sites in brain synaptic membranes treated with Triton X-100. AB - Specific binding activity of radiolabeled L-glutamic acid, a putative central excitatory neutrotransmitter, was drastically increased with increasing concentrations of Triton X-100 used for pretreatment of rat brain synaptic membranes. The binding in these Triton-treated membranes was a protein dependent, inversely temperature-dependent, stereospecific, structure-selective and saturable process with a high affinity for the amino acid. The binding activity was invariably inhibited by agonists and antagonists for the N-methyl-D-aspartic acid (NMDA)-sensitive subclass, but not by agonists for the other subclasses of excitatory amino acid neurotransmitter receptors in the brain. Scatchard analysis revealed that the binding sites consisted of a single component with a Kd of 24.4 +/- 2.5 nM and a Bmax of 0.94 +/- 0.09 pmol/mg protein. Some endogenous tryptophan metabolites such as kynurenic acid and quinolinic acid also inhibited the binding. These results suggest that synaptic membranes may indeed contain the NMDA-sensitive receptors which are disclosed by Triton X-100 treatment. PMID- 2567185 TI - The substrate and sequence specificity of the AMP-activated protein kinase. Phosphorylation of glycogen synthase and phosphorylase kinase. AB - In addition to acetyl-CoA carboxylase and HMG-CoA reductase, the AMP-activated protein kinase phosphorylates glycogen synthase, phosphorylase kinase, hormone sensitive lipase and casein. A number of other substrates for the cyclic AMP dependent protein kinase, e.g., L-pyruvate kinase and 6-phosphofructo-2 kinase/fructose-2,6-bisphosphatase, are not phosphorylated at significant rates. Examination of the sites phosphorylated on acetyl-CoA carboxylase, hormone sensitive lipase, glycogen synthase and phosphorylase kinase suggests a consensus recognition sequence in which the serine residue phosphorylated by the AMP activated protein kinase has a hydrophobic residue on the N-terminal side (i.e., at -1) and at least one arginine residue at -2, -3 or -4. Substrates for cyclic AMP-dependent protein kinase which lack the hydrophobic residue at -1 are not substrates for the AMP-activated protein kinase. PMID- 2567186 TI - Cytoskeletal reorganization during electric-field-induced fusion of Chinese hamster ovary cells grown in monolayers. AB - Mammalian cells were shown to fuse after direct electric pulsation of the plated cells in culture. The extent of fusion was controlled by the duration of the post pulse incubation. Formation of polynucleated cells was slow, even at 37 degrees C. Pre-pulse incubation with colchicine increased the fusion yield slightly. Cytoskeletal organization during the post-pulse incubation was observed using immunofluorescence techniques. Microfilaments were unaffected, but microtubules disappeared during the first minutes following the pulse, and then reformed on subsequent incubation. PMID- 2567187 TI - The Malmo polymorphism of factor IX: establishing the genotypes by rapid analysis of DNA. AB - A DNA polymorphism in the coding region of coagulation factor IX--potentially valuable for carrier detection, prenatal diagnosis, and population studies--was described in 1985. It had been discovered with monoclonal antibodies that distinguish between threonine and alanine as the 148th residue of the peptide. Its use as a diagnostic tool has been limited because threonine-containing factor IX (Malmo A) is dominant to alanine-containing factor IX (Malmo B) in immunoassays of plasma; therefore, detection of Malmo heterozygotes is not possible in all instances. A DNA method for recognizing all heterozygotes has been developed, but it also has limitations. We report the development of another DNA procedure based on amplification of the relevant DNA with the polymerase chain reaction (PCR). This method is quick, avoids the use of isotopes and x-ray film, and specifically identifies all the Malmo genotypes: hemizygotes, homozygotes, and heterozygotes. The procedure can be performed satisfactorily on small samples of blood (less than 1 mL) as suggested by Kogan et al (N Engl J Med 317:985, 1987). The method described is applicable to any genetic polymorphism that overlaps a restriction enzyme recognition site. PMID- 2567188 TI - Partial gene deletion in a family with factor X deficiency. AB - The presence of gene lesions in coagulation factor X (FX, Stuart factor) was investigated in patients with FX deficiency or an FX abnormality (FX Friuli). The proposita had a heterozygous partial deletion of the FX gene with severe deficiency of FX activity and antigen. The lesion, which was inherited from her mother, removes the 3' portion of the gene coding for the catalytic domain of the factor. In this family, two differently affected FX genes are present, leading to double heterozygosity of the proposita and thus excluding consanguinity of parents. An apparently normal gene structure was observed in the other patient with FX abnormality, suggesting the presence of a small gene lesion. PMID- 2567189 TI - Molecular basis of Sp alpha I/65 hereditary elliptocytosis in North Africa: insertion of a TTG triplet between codons 147 and 149 in the alpha-spectrin gene from five unrelated families. AB - Hereditary elliptocytosis in North Africa is frequently associated with the alpha I/65 spectrin variant, characterized by an abnormal alpha I 65-kD instead of the normal alpha I 80-kD peptide following limited trypsin digestion of whole spectrin. A similar variant (although it yielded a 68-kD fragment) has been shown recently, in two black patients, to result from the insertion of a leucyl residue at position 148 (Marchesi et al: J Clin Invest 80:191, 1987). In order to determine if the underlying molecular defect was the same in North Africans and blacks (who originate from both sides of the Sahara Desert), we performed analysis directly at the DNA level. Starting from the DNA of an Algerian alpha I/65 heterozygote in whom the mutation was associated with identifiable RFLPs, we cloned and sequenced the alpha-spectrin gene region, which includes the mutation. We thus identified an extra leucine codon (TTG) between codons 147 and 149, the coding sequence becoming CAG TTG TTG CTG instead of CAG TTG CTG. We then used the polymerase chain reaction (PCR) method and dot-blot hybridization of the amplified DNA with mutant and normal allele-specific oligonucleotides to screen the DNA from four other unrelated North African subjects with Sp alpha I/65 hereditary elliptocytosis. In all families we studied, these subjects were heterozygous for the TTG insertion. These results demonstrate that Sp alpha I/65 hereditary elliptocytosis has the same molecular basis in North Africans and blacks. PMID- 2567190 TI - Trial of brief intermittent neuroleptic prophylaxis for selected schizophrenic outpatients: clinical outcome at one year. AB - A study was conducted to investigate a novel approach to the prophylaxis of schizophrenic relapse. The treatment strategy comprised brief intermittent courses of neuroleptic agents begun as soon as non-psychotic symptoms believed to be early signs of relapse appeared. Fifty four stable, remitted outpatients meeting the American Psychiatric Association's DSM-III criteria for schizophrenia were randomised double blind to receive brief intermittent treatment with either active or placebo depot neuroleptic injections. Only three patients given placebo injections and two controls were admitted to hospital during one year of follow up. Eight (30%) of the patients given placebo injections and only 2 (7%) of the controls, however, had a recurrence of schizophrenic symptoms. Patients given placebo injections experienced fewer extrapyramidal side effects and showed a trend towards a reduction in tardive dyskinesia. Dysphoric and neurotic symptoms were identified before eight out of 11 relapses, and these symptoms were more frequent in patients given placebo depot injections. These results suggest a viable but not necessarily better alternative to continuous oral or depot treatment for less ill, chronic, stabilised schizophrenics based on the early treatment of putative prodromal symptoms of relapse. PMID- 2567191 TI - Peripheral blood stem cell autotransplantation in treatment of childhood cancer. AB - The levels of circulating hematopoietic progenitor cells were measured sequentially in eight children receiving chemotherapy for acute leukemia or neuroblastoma. Significant increases in the progenitor levels (up to 50-fold in CFU-GM numbers) were observed during post-chemotherapy cytopenia in all cases, but differences among individuals in the kinetics of recovery of less committed progenitors (CFU-mix) contrasted with the synchronized-mode of expansion of committed progenitors (CFU-GM). Peripheral blood cells were collected by repeated continuous-flow leukaphereses from three of the children during post-chemotherapy expansion of the progenitor pool and were cryopreserved after fractionation procedures. Infusion of these stored cells into the patients after marrow ablative chemotherapy established trilineage hematopoiesis. This use of stem cell rescue should be useful as an alternative to bone marrow transplantation and extends the application of cure-oriented salvage therapy to childhood cancers. PMID- 2567193 TI - Giant cell (temporal) arteritis following polyarteritis nodosa. PMID- 2567192 TI - In what circumstances is it justifiable to prescribe concomitant misoprostol and/or H2-receptor blockers with non-steroidal anti-inflammatory drugs? PMID- 2567194 TI - Fatal hypersensitivity reaction to sulphasalazine. PMID- 2567195 TI - Histopathological changes in adult cryptorchid testes. AB - The histological specimens of 73 patients who presented with undescended testes after puberty were re-evaluated. None of the specimens revealed carcinoma in situ but in the specimens of 5 patients severe atypia was detected. None of the patients, including 5 with severe atypia, has shown any evidence of invasive testicular tumour during follow-up. PMID- 2567196 TI - Biochemical properties of monoamine-rich human neuroblastoma cells. AB - The biochemical, pharmacological and immunological characterization of cells derived from human neuroblastoma tumors recently acquired great interest, since these cells may be a putative donor source for transplantation in animal models of neurological disorders. We measured monoamine levels, tyrosine hydroxylase (TH) immunostaining, and the expression of major histocompatibility cell surface antigens (MHC) in 7 human neuroblastoma cell lines. Three cell lines (LAN5, NB69 and CHP126) had high levels of monoamines. TH immunostaining was strongly positive in CHP126 and LAN5, and NB69. MHC were not detected in any of the cells with high catecholamine levels. Treatment with neuroleptics increased the metabolism of dopamine in LAN5 but not in NB69. The implantation of LAN5 cells in immunocompetent, unilaterally 6-hydroxydopamine-lesioned rats decreased the apomorphine-induced contralateral rotation. The effect of the implant was greatest in animals in which LAN5 neuroblastoma cells, pretreated with dibutyryl cyclic adenosine monophosphate (DBcAMP) and prostaglandin E1 (PGE1, were implanted into the cerebral ventricle ipsilateral to the lesion, and then irrigated with DBcAMP administered through a totally implanted drug delivery system. The effect of the implant decreased after the second week. Neuroblastoma cells were found in approximately 50% of the implanted animals. TH immunostaining was weak or absent in the grafted animals. Inflammatory changes were present in the majority of the brains examined. Extensive tumor growth was present in one animal implanted with untreated cells. Grafting of cells treated with DBcAMP and PGE1 plus with mitomycin C and bromodeoxyuridine in animals immunosuppressed with cyclosporin A reduced the apomorphine-induced rotation to 40-60% of baseline levels and this reduction persisted beyond the period of infusion with DBcAMP. Intraventricular infusion of DBcAMP in animals injected with cell culture medium produced a transient reduction of rotation to 70% of baseline. The amphetamine induced rotation was not significantly reduced during the 4 weeks follow up. Atypical cells, consistent with surviving neuroblastoma cells, were observed in the brain of all transplanted animals. TH immunostaining was weak or negative in most cases. Human neuroblastoma cells may be an alternative donor tissue for the study of the effects of transplantation in animal models of Parkinson's disease. PMID- 2567198 TI - A slow NMDA-mediated synaptic potential underlies seizures originating from midbrain. AB - After bath-perfusion with gamma-aminobutyric acid antagonists, slices of the rat's inferior colliculus were studied electrophysiologically. Synchronized epileptiform events were found to occur. The most prominent intracellular event was a sustained 30 mV depolarization which was pharmacologically and electrophysiologically characterized as an N-methyl-D-aspartate-mediated event. We propose that elicitation of this slow synaptic potential is the a priori basis of seizures arising in this midbrain nucleus. PMID- 2567197 TI - Excitatory amino acids enhance dissociation of zinc from soluble protein in cytosol of rat hippocampus. AB - An ultra-filtration method for separating free Zn from its bound form with macromolecules was used to study the effect of excitatory amino acids on dissociation of Zn in the soluble fraction of rat hippocampus. L-Glutamate, L aspartate and L-cysteic acid but not D-glutamate significantly increased the dissociated Zn at the concentration of 100 microM. 2-Amino-5-phosphonovalerate did not inhibit the effect of L-glutamate. Likewise, the stimulatory effect of N methyl-DL-aspartate and quisqualate was observed at the concentration range between 1 and 100 microM. The effect of kainate was much less potent. Taurine and GABA had no stimulatory effect on the dissociation of Zn. PMID- 2567199 TI - [3H]MK-801 binding sites in neonate rat brain. AB - [3H]MK-801 binding sites are present in neonate rat brain as early as 3 days after birth. Immature hippocampus and cortex contain approximately one sixth the concentration of binding sites of the adult, while brainstem concentration is twice as high as that of adult. [3H]MK-801 binding is stimulated by glutamate and glycine and blocked by phencyclidine and Mg2+ both in 7-day-old neonate and adult, indicating that as early as 7 days postnatally, the N-methyl-D-aspartate type glutamate receptor and MK-801 binding site are functionally coupled. PMID- 2567200 TI - Ketamine protects cultured astrocytes from glutamate-induced swelling. AB - Effects of ketamine on glutamate-induced swelling of astrocytes in primary cell culture were studied. Following the exposure to 1 mM glutamate (Glu) for 4 h the intracellular water space (as measured by 3-O-methyl-[14C]glucose uptake) of astrocytes was increased by two-fold concomitant with cell swelling and disappearance of cellular processes observed by phase-contrast microscopy. Ketamine, when co-incubated with Glu, reduced the astrocytic swelling in a dose dependent manner. These data suggested that ketamine, in addition to its function as a non-competitive N-methyl-D-aspartate receptor antagonist, is also involved in protecting astrocytes from Glu-induced swelling. PMID- 2567201 TI - [Purification to apparent homogeneity of the somatostatin receptor of the human cell line HGT-1 of gastric origin]. AB - This communication reports the isolation and the purification of the gastric somatostatin receptor from the human cell line HGT-1. The receptor has been extracted from the cell membrane by Triton X 100, and a monoclonal antibody to this was prepared. A series of affinity chromatographies (Sepharose-antibody and Sepharose-somatostatin-14) and a final purification by steric exclusion on high performance liquid chromatography columns (HPLC) allowed us to obtain a fraction enriched 20,000 fold in 125I-Tyrll-somatostatin-14 specific binding (apparent dissociation constant: 7.6 x 10(-8) M). This fraction corresponded to a molecular mass of about 90 kDa (in presence of detergent) and to a maximal binding capacity of more than 10,000 pmol/protein. It therefore has a theoretical homogeneity close to 100%. PMID- 2567202 TI - [Addition of a morphinomimetic to the continuous perfusion of 0.125% bupivacaine for peridural obstetrical anesthesia. A comparative study of fentanyl and alfentanyl]. AB - Morphinic drugs added to epidural local anesthetic during labour enhance analgesia and obstetrical conditions. Fentanyl, 1 microgram/kg-1, is safe for the newborn. Alfentanil is of faster and shorter duration and its pharmacokinetics suggests less accumulation than fentanyl. The aim of this study is to compare Alfentanil versus Fentanyl when added to an epidural continuous bupivacaine 0.125% infusion. Two groups of parturients are constituted: group A 10 micrograms/kg alfentanil, group F 1 microgram/kg fentanyl. Pain is assessed with a 0 to 10 points scale. There are no differences between the two groups for age, weight, parity, term, initial cervical dilatation and new born weight. Analgesia begins quickly in the two groups, and is more pronounced in the group A (than in the group F (p less than 0.005). Analgesia is maintained for the whole dilatation course. Pain scores increase during expulsion but are significantly lower than the initial scores. No difference is noted as regards analgesia supplementation. Obstetrical data: labour duration, oxytocin dosage, expulsion strength, instrumental extraction rate and uterin evacuation are similar in the 2 groups. No cesarean section is observed. Neonatal status, established according to Apgar scores and then Amiel Tison neurological scales (0 to 30) respectively at 30 to 120 minutes are in the same favorable ranges: Apgar score is in all cases more than 9. The neurological score is 24 (group A) and 22.9 (group F) at 30 minutes and increases significantly at 120 minutes in the 2 groups (27 in the two groups).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567203 TI - Myocardial infarct expansion during indomethacin or ibuprofen therapy for symptomatic post infarction pericarditis. Influence of other pharmacologic agents during early remodelling. AB - Evidence of acute infarct expansion and the frequency of the acute infarct expansion syndrome (acute infarct dilatation and thinning associated with hypotension and left ventricular failure but no evidence of new necrosis) occurring at two days or more after a first acute Q-wave myocardial infarction were studied using serial two-dimensional echocardiography in 221 consecutive patients (100 anterior, 121 inferior). Patients with symptomatic pericarditis were treated with indomethacin (group 1, n = 73) or ibuprofen (group 2, n = 49) and those without symptomatic pericarditis received neither drug (group 3, n = 99). The overall frequency of the acute infarct expansion syndrome was 13% and 69% of these were among the pericarditis groups. The syndrome was significantly more frequent in group 1 (22%) than group 2 (8%) (P less than 0.05) or group 3 (9%) (P less than 0.025). Serial echocardiograms revealed more expansion with greater percentage increase in the infarct containing segment length in group 1 than group 2 or group 3 (18% versus 9% versus 9%, P less than 0.005). However, the decreases in infarct segment thickness were similar in groups 1 (24%) and 2 (25%) but greater (P less than 0.001) than in group 3 (7%). Despite similar infarct size and infarct thinning in groups 1 and 2, the degree of infarct expansion was greater and the infarct expansion syndrome more frequent in group 1. However, when allowance was made for the potential protective effect of prior use of intravenous nitroglycerin and concomitant use of nifedipine, indomethacin and ibuprofen had similar effects on expansion. Thus, indomethacin or ibuprofen should be used with caution after Q-wave infarction so as to avoid further expansion. The fact that short term use of other drugs might modify infarct remodelling should be considered in studies attempting to assess efficacy of one particular drug.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2567204 TI - Presence of membrane binding sites for [D-TRP6]-luteinizing hormone-releasing hormone in experimental pancreatic cancer. AB - Characteristics of binding sites (dissociation constant: Kd and maximal binding capacity: Bmax) for [D-Trp6]-luteinizing hormone-releasing hormone [( D-Trp6]-LH RH]), somatostatin (SS-14) and epidermal growth factor (EGF) were evaluated in membrane fractions of N-Nitrosobis (2-oxopropyl) amine (BOP)-induced pancreatic adenocarcinoma of hamsters. Intact, normal hamster pancreata did not show any binding sites for [D-Trp6]-LH-RH, but specific [D-Trp6]-LH-RH binding sites with low affinity and high capacity were found after pancreatic cancer was induced with BOP. Membrane binding sites for SS-14 and EGF, with high affinity and low capacity were present, both in normal and cancerous pancreata. Normal hamster pancreatic tissue had significantly higher levels of SS-14 binding sites and lower concentration of EGF binding sites as compared to pancreatic carcinoma. In vivo treatment of hamsters bearing pancreatic cancers with microcapsules of agonist [D-Trp6]-LH-RH and the somatostatin analog RC-160 alone, or in combination, caused histopathological regression of tumors and concomitantly decreased the Kd and Bmax of [D-Trp6]-LH-RH, and increased the Bmax of the SS-14 binding sites. These findings represent the first demonstration of binding sites for [D-Trp6]-LH-RH in pancreatic cancers. Our results also suggest that tumor inhibitory effects of [D-Trp6]-LH-RH and RC-160 in pancreatic cancer could be mediated not only indirectly through suppression of sex-steroids, gastrointestinal hormones and growth factors, but also directly by an action on specific binding sites located on the tumor membranes. PMID- 2567206 TI - Loss of heterozygosity on the short arm of chromosome 3 in carcinoma of the uterine cervix. AB - Loss of genes at specific chromosomal loci is a common genetic alteration in human tumors and is thought to be critical for unmasking the recessive genetic changes for tumorigenesis. To learn whether such recessive mutations are involved in the development of carcinoma of the uterine cervix, 18 fresh tumors were analyzed by Southern blot hybridization using 34 polymorphic DNA markers covering 19 different chromosomes. We found loss of heterozygosity at the D3S2 locus on chromosome 3p in all nine patients who could be evaluated. Human papillomavirus type 16 and type 18 were present in seven and three of 18 tumors, respectively, while no amplification of 13 oncogenes, including c-myc and H-ras, was detected in these tumors. These results suggest that recessive genetic changes on chromosome 3p are one of the important genetic alterations for the development of carcinoma of the uterine cervix. Since this locus is also lost commonly in lung cancer and in renal cell carcinoma, it is possible that these three different types of adult tumors result from mutations of the same recessive gene on chromosome 3p. PMID- 2567205 TI - Factors responsible for variable reported lineages of HL-60 cells induced to mature with butyric acid. AB - HL-60 is a multipotential human leukemia cell line widely used as an in vitro model to investigate myeloid differentiation. A variety of compounds can reproducibly induce these cells to differentiate towards specific lineages. However, under what appear to be similar experimental conditions, various laboratories have reported either neutrophilic differentiation or monocytic differentiation after butyric acid induction. We investigated different hypotheses to explain these dissimilar findings. First, the potential role of variable 1,25-dihydroxyvitamin D3 (VD3) concentrations in commercial fetal calf serum was assessed. Second, possible differences between laboratories inherent to the HL-60 cells themselves were explored. Lineage was assessed by morphology, histochemistry (nonspecific esterase), and neutrophil-specific (CD15) and monocyte-specific (MO2) surface antigens. We found that increasing concentrations of VD3 spanning the range reported in commercial fetal calf serum (25 to 155 pg/ml) act in synergy with butyric acid to result in higher monocyte/neutrophil ratios at the higher VD3 concentrations. Different lots of serum led to monocyte/neutrophil ratios in proportion to their VD3 concentrations. Starting with HL-60 cells obtained from different laboratories, several single-cell clones were derived which yielded either high percentages of monocytes, high percentages of neutrophils, or intermediate mixes of both cell types after induction with butyric acid. We conclude that the wide variation of VD3 concentration found in different lots of commercial fetal calf serum and intrinsic differences in HL-60 cells are two identifiable factors that can explain the discrepancies in lineage observed by different investigators after butyric acid induction of HL-60 cells. PMID- 2567207 TI - Enhancement of the effectiveness of Lyt-2+ T-cells for adoptive chemoimmunotherapy by short-term exposure of tumor-bearer spleen cells to polyethylene glycol and/or melphalan. AB - Uncultured tumor-infiltrated spleen cells (TISpC) from mice bearing large (20-22 mm) s.c. MOPC-315 plasmacytomas were previously shown to be ineffective in bringing about the cure of mice bearing a nonpalpable (Day 4) tumor that had been treated with a subcurative dose (10 mg/kg) of cyclophosphamide (i.e., adoptive chemoimmunotherapy, ACIT) (M. B. Mokyr, J. C. D. Hengst, and S. Dray, Cancer Res., 42:974-979, 1982). Here we show that TISpC cultured for 5 days in the presence of inactivated MOPC-315 stimulator cells acquire some effectiveness in curing mice by ACIT, and this effectiveness is greatly enhanced if polyethylene glycol 6000 (PEG) is also added to the culture. The Lyt 2+ T-cells, and not the L3T4+ T-cells, are responsible for the effectiveness of the cultured TISpC in ACIT. In fact, the L3T4+ T-cells are apparently not required even during culture of TISpC for the generation of Lyt 2+ T-cells effective in ACIT. Although the TISpC cultured with MOPC-315 cells and PEG contained approximately twice as many Lyt 2+ cells as did TISpC cultured without PEG, the increase in the activity of the former cells is not due simply to the increase in the percentage of Lyt 2+ cells, but is most likely due to an increase in the percentage and/or activity of Lyt 2+ cells with specificity for MOPC-315-associated antigens. The effectiveness of TISpC cultured with MOPC-315 stimulator cells and PEG in ACIT can be enhanced even further by pretreatment of these cells with the immunomodulating agent melphalan (0.5 nmol/ml) prior to culture initiation. Thus, the above methods of culture render ineffective lymphoid cells effective in ACIT and are suitable for evaluation in protocols for human cancer therapy. PMID- 2567208 TI - Biochemically directed therapy of leukemia with tiazofurin, a selective blocker of inosine 5'-phosphate dehydrogenase activity. AB - Tiazofurin (2-beta-D-ribofuranosylthiazole-4-carboxamide, NSC 286193), a selective inhibitor of the activity of IMP dehydrogenase (EC 1.1.1.205), the rate limiting enzyme of de novo GTP biosynthesis, provided in end stage leukemic patients a rapid decrease of IMP dehydrogenase activity and GTP concentration in the blast cells and a subsequent decline in blast cell count. Sixteen consecutive patients with end stage acute nonlymphocytic leukemia or myeloid blast crisis of chronic granulocytic leukemia were treated with tiazofurin. Allopurinol was also given to inhibit xanthine oxidase activity to decrease uric acid excretion and to elevate the serum concentration of hypoxanthine, which should competitively inhibit the activity of hypoxanthine-guanine phosphoribosyltransferase (EC 2.4.2.8), the salvage enzyme of guanylate synthesis. Assays of IMP dehydrogenase activity and GTP concentration in leukemic cells provided a method to monitor the impact of tiazofurin and allopurinol and to adjust the drug doses. In this group of patients with poor prognosis, five attained a complete hematological remission and one showed a hematological improvement. A marked antileukemic effect was seen in two other patients. All five evaluable patients with myeloid blast crisis of chronic granulocytic leukemia reentered the chronic phase of their disease. Five patients with acute nonlymphocytic leukemia were refractory to tiazofurin and three were unevaluable for hematological effect because of early severe complications. Responses with intermittent 5- to 15-day courses of tiazofurin lasted 3-10 months. Tiazofurin had a clear antiproliferative effect, but the pattern of hematological response indicated that it appeared to induce differentiation of leukemic cells. In spite of toxicity with severe or life threatening complications in 11 of 16 patients, tiazofurin was better tolerated in most patients than other antileukemic treatment modalities and provided a rational, biochemically targeted, and biochemically monitored chemotherapy which should be of interest in the treatment of leukemias and as a paradigm in enzyme pattern-targeted chemotherapy. PMID- 2567209 TI - In vivo antitumor effect of methotrexate conjugated to a monoclonal IgM antibody specific for stage-specific embryonic antigen-1, on MH-15 mouse teratocarcinoma. AB - Methotrexate (MTX) was coupled to an IgM monoclonal antibody specific for stage specific embryonic antigen-1 (SSEA-1), and the resulting immunoconjugate (MTX anti-SSEA-1) was used for in vivo drug targeting in mice bearing MH-15 teratocarcinoma. Immunoconjugates having an average of 65 mol MTX/mol antibody retained full antigen-binding capacity. Mice bearing well-established tumors (approx. 1 g) were treated i.v. using the immunoconjugate. MTX-anti-SSEA-1 at 15 mg/kg of drug had significant antitumor activity with no significant systemic toxicity. Neither an irrelevant isotype-matched conjugate, MTX-MOPC-104E, prepared from the MOPC 104E myeloma protein, nor free MTX injected alone or with either antibody had any significant antitumor effect. These results indicate that IgMs can be effective drug carriers for tumor targeting in spite of their high molecular mass, and that antigens that are selectively accessible in tumors, even though present in normal tissues, can be suitable targets for in vivo chemoimmunotherapy. PMID- 2567210 TI - Induction of nonspecific killer cells by delayed-type hypersensitivity against soluble protein antigens in murine peritoneal cavities. AB - We induced nonspecific killer cells in the local site of delayed-type hypersensitivity against keyhole limpet hemocyanin or ovalbumin. Delayed-type hypersensitivity was induced in the peritoneal cavities of mice, and peritoneal exudate cells (PEC) were collected. These PEC were found to have killer activity toward SP2 and YAC-1 cells (target cells susceptible to natural killer cells) by 4-h 51Cr-release assays. The induction of killer activity in PEC was observed in parallel with the eliciting of delayed-type hypersensitivity in the peritoneal cavity, in which the killer activity was maximum 24-48 h after the antigen challenge, but was not induced in nu/nu mice and was induced in an antigen specific way. These killer cells did not adhere to nylon wool and had Thy1 and asialo-GM1 antigens on their surfaces. Their precursor cells were also asialo-GM1 positive. These findings indicate that the killer cells probably belong to the NK cell lineage. Results of tumor challenge experiments showed that these killer cells had an antitumor effect in vivo as well as in vitro. PMID- 2567212 TI - Role of protein kinase C activators and inhibitors, calmodulin antagonists and membrane sialic acids in polyamine transport in murine leukemia cells. AB - The effects of activators and inhibitors of protein kinase C (phorbol esters and H-7) and antagonist to calmodulin (TFP) on polyamine transport in murine leukemia (L1210) cells are investigated. Phorbol esters and H-7 are found to enhance and curtail the uptake of 14C-Spermidine (Spd) respectively in L1210 cells. TFP also inhibits the uptake process. After desialation of cells with neuraminidase, phorbol esters are found to further increase the uptake of 14C-Spd by 35% compared to untreated cells. The sialic acid contents of the cells are regenerated by incubation with 14C-glucosamine for 18 hours. The regenerated cells mimic like untreated cells for the uptake of 14C-Spd i.e. after regeneration of sialic acids, the Spd uptake is curtailed significantly in comparison with desialated cells. Phorbol esters are found to enhance the activity of transglutaminase present in L1210 cells while H-7 and TFP exhibit reverse effects. The possible role of phorbol esters, H-7 and TFP and their effects on transglutaminase activity in relation with Spd transport process are discussed. PMID- 2567211 TI - Amelioration of experimental lung metastasis in mice by therapy with anti-CD3 monoclonal antibodies. AB - Binding of CD3-specific antibodies to the TcR-CD3 complex results in T cell activation without the need for occupation of the T cell receptor (TcR) by its ligand. Murine T cells activated in this manner will kill a broad range of tumor targets but not normal lymphoblasts. We report here that non-specific cytolytic activity can be induced in vivo by a single i.p. injection of nonlytic 145-2C11 anti-CD3 monoclonal antibody. At least three populations of effector cells are activated in these mice. These are non-MHC (major histocompatibility complex) restricted cytotoxic T lymphocytes, activated natural killer cells, and lymphokine-activated killer cells. Anti-CD3 treatment is effective in significantly reducing the number of lung tumor nodules which form in mice inoculated with oncogenic ras-transfected syngeneic 10T1/2 fibroblasts. Anti-CD3 activated killer cells may, therefore, find a future role in cancer immunotherapy. PMID- 2567214 TI - Activities of dipeptidyl peptidase II and dipeptidyl peptidase IV in synovial fluid from patients with rheumatoid arthritis and osteoarthritis. AB - We examined the activities of peptidases in synovial fluid from patients with rheumatoid arthritis (RA) and osteoarthritis (OA). Dipeptidyl peptidase IV (DPP IV) activity was lower in synovial fluid from patients with RA, in contrast to the increase of DPP II activity in synovial fluid, as compared with OA. The DPP II/DPP IV ratio for synovial fluid was significantly higher in patients with RA than in patients with OA. A significant correlation was observed between the DPP II/DPP IV ratio for synovial fluid from patients with RA and the amount of C reactive protein reaction. These results may be useful in the diagnosis of joint effusion of unknown origin. PMID- 2567213 TI - [Tardive dyskinesia after low doses of neuroleptics and attempts at treatment]. AB - On the example of a 63-year-old neurotic patient dependent on alcohol and anxiolytic drugs the author draws attention to the risk of the development of a tardive dyskinesia after the long-term administration of "neurotic doses" of neuroleptics to non-psychotic patients. The author emphasizes the greater danger in the group of patients dependent on alcohol. He describes an attempt of concurrent treatment of neurosis, tardive dyskinesia and hypertension by a non selective beta-blocker metopranolol (Trimepranol Spofa) administered for eight months. This drug was according to the author's information administered for the first time in tardive dyskinesia. It seems that it improves tardive dyskinesia in much smaller doses than anxiety and sleep disorders. PMID- 2567215 TI - Use of the polymerase chain reaction for simultaneous analysis of two Pst I polymorphisms linked to cystic fibrosis. PMID- 2567216 TI - Serum mitochondrial aspartate aminotransferase activity: not useful as a marker of excessive alcohol consumption in an unselected population. AB - Using an immunochemical method, we measured the activity of the mitochondrial isoenzyme (mAST) of aspartate amino-transferase (EC 2.6.1.1, AST) in the serum of 687 subjects attending the Centre for Preventive Medicine for a health examination. The distributions of the activities were asymmetrical, with mean values of 1.8 U/L (SD 2.0) for men and 1.4 U/L (SD 1.6) for women. The average ratio of mitochondrial to total AST activity was 0.051 (range 0-0.42). In this unselected population we found no change in the mitochondrial activity or in the mitochondrial-to-total ratio attributable to alcohol consumption, even in subjects who consumed more than 88 g per day. Of 35 men with an alcohol consumption greater than 88 g/d, 19 had a serum gamma-glutamyltransferase activity of greater than or equal to 60 U/L, 17 had glutamate dehydrogenase values greater than or equal to 5 U/L, and only nine had an mAST activity greater than or equal to 3 U/L (values corresponding to the 80th percentiles of the total population). We conclude that the test is not particularly useful as a screening procedure in an unselected population under present-day conditions of measurement. PMID- 2567217 TI - Laboratory markers of alcoholism. PMID- 2567219 TI - Partial deletions of factor VIII gene as molecular diagnostic markers in haemophilia A. AB - A panel of 27 families at risk for haemophilia A was studied by RFLP analysis using the anonymous probe St14.1 (DXS52), a cDNA probe spanning the exons 16 to 19, and a genomic fragment of intron 22. In two patients with severe haemophilia A, who did not form inhibitors, abnormal RFLP patterns were found, that can be interpreted as partial deletions in exons 17 to 19, and intron 22, respectively. In a case with moderate haemophilia A a further partial deletion in intron 22 was found. The significance of the deletions detected as markers for pedigree analysis and prevention of haemophilia A is demonstrated. PMID- 2567218 TI - Lipid effects of celiprolol, a new cardioselective beta-blocker, versus propranolol. AB - The metabolic effects of celiprolol, a new beta-adrenoceptor blocking agent with intrinsic sympathomimetic activity and alpha 2-blocking properties, were evaluated in a series of patients with hypertension, both with and without hyperlipidemia. Propranolol was tested as the reference drug in a randomized double-blind trial. Of the 35 patients of both sexes who completed the study, 17 were hyperlipidemic (low-density lipoprotein cholesterol greater than or equal to 170 mg/dl) and 18 were normolipidemic. Both drugs exerted a similar hypotensive effect after gradual dose adjustment; however, propranolol reduced heart rate to a higher extent (-20.5%) than celiprolol (-7.7%). Propranolol determined a significant rise of total and very low-density lipoprotein (VLDL) associated triglyceridemia, whereas high-density lipoprotein cholesterol (HDL cholesterol) levels and the total cholesterol/HDL cholesterol ratios were significantly depressed, particularly in hyperlipidemic patients. Celiprolol, in contrast, slightly decreased triglyceridemia (significantly in the hyperlipidemic group at week 12) and caused a 5% increase of the HDL cholesterol levels. The total cholesterol/HDL cholesterol ratio was reduced by celiprolol at week 16 in both hyperlipidemic and normolipidemic patients. The effects of the two beta adrenoceptor blockers on HDL cholesterol and triglyceride levels differed significantly after 12 and 16 weeks of treatment, which confirm the divergent metabolic effects of the two agents. PMID- 2567220 TI - Choline transport specificity in animal cells and tissues. AB - 1. Beta carbolines inhibit choline transport in rat brain. 2. The aziridinium ring on the nitrogen of mustard analogs of choline causes irreversible binding to the carrier in rat brain. 3. The uptake system in rat brain is stereoselective, requires a quaternary nitrogen, and prefers analogs with a nitrogen-oxygen distance of about 3.26 A. 4. In mouse brain troxonium derivatives inhibit choline transport. 5. In cuttlefish optic lobes and torpedo electric organ pyrene derivatives potently inhibit choline transport. 6. In guinea pig placenta, the affinity of the choline carrier remains high even when this molecule lacks one or two methyl groups. PMID- 2567221 TI - Characterization of pre- and postsynaptic dopamine receptors in Lymnaea. AB - 1. The effects of dopamine and several synthetic agonists and antagonists were studied using two identified neurons of the snail Lymnaea stagnalis. 2. In both the buccal-2 (B-2) neurons and the pedal giant (RPeD1) neuron dopamine elicited a hyperpolarizing response at least partly due to potassium efflux. RPeD1 is itself dopaminergic, implicating autoreceptors in its response to dopamine. 3. The following agents were tested: agonists--LY171555, pergolide, SKF38393, (-)-3-PPP, R(-)NPA and dopamine; antagonists--SCH23390, sulpiride, and metaclopramide. Dibutyryl cAMP was applied to determine whether the response is cAMP-mediated. 4. Results indicate that the pharmacological profiles of dopamine receptors on these neurons are inconsistent with those of either D-1, D-2 or autoreceptors in mammals. PMID- 2567222 TI - Acute toxicity of combinations of sodium dichromate, sodium arsenate and copper sulphate in the rat. AB - 1. The intraperitoneal treatment of adult male Wistar rats with various combinations of low doses of sodium dichromate (5 mg/kg), sodium arsenate (25 mg/kg) and copper sulphate (5.9 mg/kg) tended to counteract the inherent acute toxicity of each compound. 2. The co-administration of low doses of one or more of the test compounds with a high dose of sodium dichromate (35 mg/kg), sodium arsenate (90 mg/kg) or copper sulphate (23.5 mg/kg) resulted in a significant increase in acute toxicity in comparison with that produced by the administration of high doses of dichromate, arsenate or Cu2+ alone. PMID- 2567224 TI - Some selected peripheral blood and haemopoietic system indices in Wistar rats with chronic vanadium intoxication. AB - 1. Wistar rats of both sexes received vanadium in drinking water in the amount of 23-29 mg/kg body weight in the form of ammonium metavanadate (AMV) for a period of 2, 4 and 8 weeks. 2. Animals treated in this way ate less food and drank less AMV solution as compared with the amount of water consumed by the controls; they suffered from diarrhoea, and owing to this the increment in body weight was reduced. 3. Vanadium decreased erythropoiesis and maturation of red blood cells, which was expressed by a reduced erythrocyte count and haemoglobin level and increased reticulocyte and polychromatophilic erythrocyte count in the peripheral blood. 4. The composition percentage of the bone marrow cells and the peripheral blood leukocyte count did not undergo noticeable changes under the influence of vanadium. PMID- 2567223 TI - The actions of verapamil at the neuromuscular junction. AB - 1. The actions of the calcium channel blocker verapamil were studied at the neuromuscular junction of the frog Rana pipiens. 2. In the presence of 50 microM verapamil, subthreshold endplate potentials were produced, and the quantal content was reduced by a factor of 3. 3. Verapamil (10-50 microM) also reduced the postjunctional membrane sensitivity as measured by (a) carbachol iontophoresis and (b) miniature endplate potential amplitude. In addition, verapamil had a strong inhibitory effect on the postjunctional membrane response to repetitive iontophoretic application of carbachol. 4. Thus, verapamil has both pre- and postsynaptic actions at the neuromuscular junction. PMID- 2567225 TI - The effect of T-2 toxin on active sodium transport across frog skin in the presence of ADH and amphotericin B. AB - 1. The effect of T-2 toxin on active sodium transport across frog skin both in the presence and in the absence of stimulants of sodium transport, such as Amphotericin B and ADH, was studied using the short circuit current technique with the following results. 2. T-2 toxin produces inhibition of active sodium transport in a dose-response correlation. 3. This effect is irreversible since the washing out of the tissue does not restore its functionality. This indicates that the micotoxin may cross the cellular membrane and act on the internal site. 4. ADH partially removes the inhibitory effect of T-2 toxin. 5. The increase of the sodium pool in the cell as determined by Amphotericin B does not reverse the inhibitory effect of T-2 toxin. 6. The biological significance of these data is discussed in regard to the possible effect of T-2 toxin on Na+, K+-ATPase activity either directly or by a reduction in the metabolic supply of substrates, or by a modified stoichiometry of the pump reaction. PMID- 2567226 TI - Effect of bromobenzene and O-bromophenol on kidney and liver of English sole (Parophrys vetulus). AB - 1. English sole (Parophrys vetulus) were injected intraperitoneally with a single dose of 9.8 mmol bromobenzene/kg of fish or 1.9 mmol O-bromophenol/kg of fish, both known renal toxicants in mammals. 2. Kidney, liver, gill spleen, intestines, heart and blood samples were subsequently obtained up to 48 hr post-injection for determination of microscopic lesions, concentrations of selected tissue antioxidants (glutathione and ascorbic acid), and selected serum parameters. 3. Bromobenzene and O-bromophenol were both found to be hepatotoxic in English sole, as indicated by the presence of hepatocellular coagulation necrosis and fatty change in the liver, altered glutathione and ascorbic acid levels in liver tissue, elevated serum aspartate aminotransferase and alkaline phosphatase activity and increased serum glucose and triglyceride levels. 4. No evidence of nephrotoxicity was found in English sole exposed to either toxicant. 5. It is concluded that bromobenzene and O-bromophenol cannot be used as model nephrotoxicants but can be used as hepatotoxicants in English sole. PMID- 2567227 TI - Differential effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin on adipose tissue lipoprotein lipase activity in the guinea pig, rat, hamster, rabbit, and mink. AB - 1. Adipose tissue lipoprotein lipase (LPL) activity of several different animal species was determined after i.p. administration of 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD). 2. TCDD caused a significant reduction in LPL activity and an increase in serum triglyceride concentration in guinea pigs, rabbits, and hamsters but not rats. 3. TCDD increased adipose tissue LPL activity of mink and lowered their serum triglyceride concentration. 4. Results of this study indicate that profound differences occur in lipid metabolism between various species in response to TCDD and these changes do not appear to be related to generalized toxicity such as wasting. PMID- 2567228 TI - Comparative actions of salicylate on the amphibian lateral line and guinea pig cochlea. AB - 1. Salicylate actions on afferent nerve activity in the Xenopus lateral line and on cochlear potentials in guinea pig were investigated. 2. In the lateral line, salicylate (0.3-2.5 mM) suppressed spontaneous activity, water motion evoked excitation and responses to L-glutamate (1-2 mM) and kainate (10-20 microM). 3. In the guinea pig, salicylate (0.6-10 mM) suppressed the compound action potential (CAP) and increased N1 latency at low but not high sound intensities. 4. In the lateral line salicylate action may involve an antagonism of the hair cell transmitter on the afferent nerve. 5. In the cochlea salicylate may suppress the active process or cochlear amplifier. PMID- 2567229 TI - Gender differences in the brain: are they relevant to the pathogenesis of schizophrenia? AB - Gender differences are present in the clinical expression of schizophrenia, age of onset, course of illness, and response to pharmacologic treatment. These differences are not surprising in view of the normal gender differences in brain growth, differentiation, adult brain structure, and neurochemistry. The present review examines what is presently known about brain gender differences, and whether this information is consistent with the published reports of brain functional and morphological abnormalities in schizophrenia. Whether gender differences in the brain can explain the gender differences in clinical aspects of the disorder remains unknown. PMID- 2567230 TI - A case of multiple endocrine neoplasia (MEN) type 1; the immunohistochemical and ultrastructural studies of its tumors and the analysis of hormones in tumor extracts. AB - We reported a case of sporadic multiple endocrine neoplasia type 1, with multiple insulinoma, parathyroid adenoma, and pituitary tumor. Measurement of hormone contents and immunohistochemical studies of the pancreatic tumors showed that the tumors contained insulin, glucagon, somatostatin, and pancreatic polypeptide. Furthermore, the concentrations of these hormones were different in each tumor. Insulin extracted from the pancreatic tumors analyzed by reversed-phase high performance liquid chromatography revealed no structural abnormalities. On the other hand, in gel filtration evaluation of the extract of the parathyroid adenoma, it was found that the tumor extract contained a macromolecular parathyroid hormone (molecular weight 20,000 to 25,000). PMID- 2567232 TI - Subcellular localization and kinetic properties of phosphatidylinositol 4,5 bisphosphate phospholipase C and inositol phosphate enzymes from human peripheral blood mononuclear cells. AB - Peripheral blood mononuclear cells from normal donors exhibited phosphatidylinositol 4,5-bisphosphate phospholipase C (PIP2-PLC), inositol 1,4,5 trisphosphate (IP3) and inositol 1-phosphate (IP)-monophosphatase activities which were mostly recovered in the cytosol fraction. In both cytosol and particulate fractions PIP2-PLC displayed the highest activity at pH 6.2, whereas IP3 and IP-monophosphatases showed the same optimal pH at 7.0. While the PIP2-PLC displayed close apparent Km values in cytosol and particulate fractions, both inositol-monophosphatases were found to show substrate affinities for IP and IP3 characteristic of these two fractions, with an higher affinity in the soluble fraction. PMID- 2567231 TI - Activity of thyroid stimulating antibody and thyroid stimulation blocking antibody determined by radioiodine uptake into FRTL-5 cells. AB - To investigate the pathophysiology of patients with autoimmune thyroid diseases, we measured serum thyroid stimulating antibody (TSAb) activity and thyroid stimulation blocking antibody (TSBAb) activity by determining the radioiodine (125I) uptake into FRTL-5 cells. FRTL-5 cells were pre-incubated for seven days with 5H medium and then incubated for 48 hours with patients' crude IgG prepared by polyethylene glycol precipitation. In order to measure TSBAb, 10 microU/ml TSH was also added. 125I was added one hour before the end of the 48 hour incubation period. After the incubation, the medium was aspirated, and the radioactivity in the cells was counted. In patients with untreated hyperthyroid Graves' disease, TSAb was detectable in 18 of 20 patients, the detectability being 90%, and activity showed a statistically significant positive correlation with TSAb activity determined by c-AMP accumulation. Out of 41 patients with hypothyroidism, TSBAb determined by 125I uptake was positive in six cases, the detectability being 14.6%. The inhibition of 125I uptake by one of these six IgGs was suggested to be at the TSH receptor level because it inhibited TSH induced c AMP accumulation and showed positive thyrotropin binding inhibitor immunoglobulin (TBI I) activity, but did not inhibit the forskolin- and (Bu)2cAMP-induced 125I uptake. Inhibition of another IgG was suggested at the post-receptor level because it did not inhibit TSH induced cAMP accumulation and showed negative TBI I activity, but inhibited forskolin- and (Bu)2cAMP-induced 125I uptake.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567233 TI - The nephrotoxic potential of neomycin in the horse. AB - Neomycin was administered intramuscularly to four normal adult horses at a dose rate of 10 mg/kg bodyweight every 12 h for 10 days (21 doses). The pharmacokinetic behaviour of neomycin with multiple dosing was characterised and a range of blood chemical and urinary parameters examined for evidence of nephrotoxicity. There was evidence of physical renal tubular injury (enzymuria and cylindriuria) within four days of neomycin administration but this subsided following cessation of treatment. No significant functional nephrotoxicity was detected. More severe nephrotoxicity might be expected in ill horses and it is recommended that several clinicopathological results be monitored serially in those horses receiving parenteral neomycin. PMID- 2567234 TI - Monorchidism in the horse. AB - Six horses with monorchidism, identified at surgery for cryptorchidectomy, are reported. All six presented with a single scrotal testis. Following surgical removal of one testis, they were either hormonally, anatomically or behaviourally determined to be geldings. Three other horses reported in the literature are reviewed. Of these nine cases of monorchidism, eight were thought to be caused by testicular degeneration and one by testicular agenesis. The vaginal process was present in all of the former and absent in the latter. The left side was involved in five of these eight horses. In seven, the epididymis was absent and, in the remaining two, only the epididymal tail was present. The condition was thought to be congenital in the six horses in this series. A surgical approach to identify accurately monorchid horses is described. PMID- 2567235 TI - Cardiomyopathy in phaeochromocytoma: a case report. AB - This case report describes the anaesthetic management of a patient with bilateral phaeochromocytoma and cardiomyopathy. The hypertension and supraventricular tachycardia commonly seen during manipulation of this tumour was controlled by administration of sodium nitroprusside and verapamil. Verapamil allowed effective management of supraventricular rhythm disturbances and arterial blood pressure, and cardiac index remained unchanged during and after phaeochromocytoma removal. As the haemodynamic side-effects of the calcium blocking agent are readily reversed by intravenous calcium chloride, it may have a useful part to play in such cases. PMID- 2567236 TI - Functional hepatocyte heterogeneity. Vascular 2-oxoglutarate is almost exclusively taken up by perivenous, glutamine-synthetase-containing hepatocytes. AB - 1. In isolated perfused rat liver maximal rates of 2-[1-14C]oxoglutarate uptake were about 0.4 mumol.g-1 .min-1; half-maximal rates of 2-[14C]oxoglutarate uptake were observed with influent concentrations of about 100 microM. 2 [14C]Oxoglutarate uptake by the liver was not affected by the direction of perfusion, but was decreased by about 80-90% when Na+ in the perfusion fluid was substituted by choline+, suggesting a Na+-dependence of hepatic 2-oxoglutarate uptake. In the absence of added ammonia, [14C]oxoglutarate uptake by the liver was about twice the net oxoglutarate uptake, indicating a simultaneous release of unlabeled oxoglutarate from perfused rat liver. 2. 14C-Labeled metabolites derived from [1-14C]oxoglutarate and recovered in the effluent perfusate were 14CO2 and 14C-labeled glutamate and glutamine; they accounted for 85-100% of the radiolabel taken up by the liver. 14CO2 was the major product (more than 70%) from [1-14C]oxoglutarate taken up the liver, provided glutamine synthesis was either inhibited by methionine sulfoximine or the endogenous rate of glutamine production was below 40 nmol.g-1.min-1. 3. Stimulation of glutamine synthesis by ammonia did not affect [14C]oxoglutarate uptake by the liver, but considerably increased net hepatic oxoglutarate uptake, indicating a decreased release of unlabeled oxoglutarate from the liver. Stepwise stimulation of hepatic glutamine synthesis led to a gradual decrease of 14CO2 production and radiolabel was recovered increasingly as [14C]glutamine in the effluent. At high rates of glutamine formation (i.e. about 0.6 mumol.g-1.min-1), about 60% of the [1 14C]oxoglutarate taken up by the liver was recovered in the effluent as [14C]glutamine. 14CO2 and [14C]glutamine production from added [1 14C]oxoglutarate were dependent on the rate of hepatic glutamine synthesis but not on the direction of perfusion. Extrapolation of 14C incorporation into glutamine to maximal rates of hepatic glutamine synthesis yielded an about 100% utilization of the [14C]oxoglutarate taken up by the liver for glutamine synthesis. This was again true for both the antegrade and the retrograde perfusion directions. On the other hand, addition of ammonia did not affect 14CO2 production from labeled oxoglutarate, when glutamine synthetase was inhibited by methionine sulfoximine. 4. The data suggest that vascular oxoglutarate is almost exclusively taken up by the small perivenous hepatocyte population containing glutamine synthetase, i.e. a cell population comprising only 6-7% of all hepatocytes. Thus, the findings demonstrate the existence of a, to date, uniquely zonally distributed oxoglutarate transport system which is probably Na+-dependent in the plasma membrane.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2567237 TI - A combination of intracellular leucine with either glutamate or aspartate inhibits autophagic proteolysis in isolated rat hepatocytes. AB - It has been shown previously that the inhibition of autophagic proteolysis in liver by a physiological mixture of amino acids can be mimicked completely by addition of leucine in combination with alanine [Leverve, X. M., Caro, L. H. P., Plomp, P. J. A. M. and Meijer, A. J. (1987) FEBS Lett. 219, 455-458]. We have now further defined conditions which lead to this inhibition. Isolated rat hepatocytes were incubated in the perifusion system in which the cells can be maintained at a steady state in the presence of low amino acid concentrations. Combinations of leucine (0.5 mM) with either alanine, glutamine, asparagine or proline (2 mM) inhibited proteolysis by 40-50%. Under these conditions, both in the absence and presence of the transaminase inhibitor, aminooxyacetate, a correlation was found between the extent of inhibition of proteolysis and the sum of the total intracellular amounts of aspartate and glutamate. Inhibition of proteolysis by leucine and leucine analogues did not correlate with their ability to activate glutamate dehydrogenase. PMID- 2567238 TI - Molecular characterization of the genetic risk in breast cancer. PMID- 2567239 TI - Pharmacological control of the human gastric histamine H2 receptor by famotidine: comparison with H1, H2 and H3 receptor agonists and antagonists. AB - Histamine 0.1 microM-0.1 mM increased adenylate cyclase activity five- to ten fold in human fundic membranes, with a potency Ka = 3 microM. The histamine dose response curve was mimicked by the H3 receptor agonist (R) alpha-MeHA, but at 100 times lower potency, Ka = 0.3 mM. Histamine-induced adenylate cyclase activation was abolished by H2, H1 and H3 receptor antagonists, according to the following order of potency IC50: famotidine (0.3 microM) greater than triprolidine (0.1 mM) thioperamide (2 mM), respectively. Famotidine has no action on membrane components activating the adenylate cyclase system, including the Gs subunit of the enzyme stimulated by forskolin and cell surface receptors sensitive to isoproterenol (beta 2-type), PGE2 and VIP. The Schild plot was linear for famotidine (P less than 0.01) with a regression coefficient r = 0.678. The slope of the regression line was 0.64 and differs from unity. Accordingly, famotidine showed a slow onset of inhibition and dissociation from the H2 receptor in human cancerous HGT-1 cells. The results demonstrate that famotidine is a potent and selective H2 receptor antagonist with uncompetitive actions in human gastric mucosa. Consequently, famotidine might be a suitable drug with long-lasting actions in the treatment of Zollinger-Ellison syndrome. The results also confirm and extend the previous observations that (R) alpha-MeHA and thioperamide are two selective ligands at histamine H3 receptor sites. In the human gastric mucosa, these drugs are respectively 330 and 6700 times less potent than histamine and famotidine on the adenylate cyclase system. The possible involvement of histamine H3 receptors in the regulation of gastric secretion is proposed. PMID- 2567240 TI - Oral E2 prostaglandins affect endocrine cell populations in the gastric antrum of the rat. AB - The aim of the present study was to investigate antral endocrine cell populations and tissue and circulating hormone levels following a 4-week oral regimen with prostaglandin E2 (25, 250 and 5000 micrograms/kg-1 b.i.d.) or a stable methyl analogue (5 and 50 micrograms kg-1 b.i.d.). Epithelial hyperplasia of the gastric antrum was observed with the highest dose of prostaglandin E2 and both doses of the analogue, as evaluated by stereological methods and conventional cell count. The treatments significantly affected the endocrine cell population. Somatostatin immunoreactive cells were increased in proportion to the increased epithelial cellularity and plasma levels of somatostatin were increased in parallel. The tissue content of somatostatin-like immunoreactivity differed according to the extraction procedure used, and was significantly higher than controls in specimens extracted in neutral water. In the neutral extracts an immunoreactive somatostatin of unidentified molecular structure dominated quantitatively over somatostatin 14 and 28, which were the major components in acetic acid extracts. The serotonin-immunoreactive cell population was also significantly increased by natural prostaglandin E2 and the analogue but the gastrin cell population was not significantly affected by treatments. Accordingly, no significant changes were observed in tissue or plasma gastrin levels. It is concluded that the epithelial hyperplasia of the antral epithelia produced by E2 prostaglandins is associated with selective changes of endocrine cell populations. The changes were proportional to the increases of epithelial cellularity and required quantitative determination of the total antral volume to be detected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567241 TI - A monoclonal antibody to murine CD45R distinguishes CD4 T cell populations that produce different cytokines. AB - CD4 T cell clones have been shown to be functionally heterogeneous in the mouse. However, it is not known if normal CD4 T cells are also functionally heterogeneous, or whether functional specialization is a result of cloning and long-term culture. To approach this question, a monoclonal antibody reacting with a subset of CD4 T cells has been prepared by immunization of rats with different cloned T cell lines all sharing the same functional activity. This monoclonal antibody reacts with a subset of CD45 (T200) molecules by binding to a determinant requiring the expression of the second variable exon of the CD45 molecule. Some CD4 T cells bear high levels of this marker, while others react only weakly. This antibody was used to separate CD4 T cells into two subpopulations. The brightly staining population was found to produce interleukin (IL) 2 and not IL 4, while the weakly staining population produced IL 4 and not IL 2. These data demonstrate that CD4 T cells in normal mice are already functionally committed, and that they differentially express forms of CD45 that contain the second variable exon. PMID- 2567242 TI - Masking of veto function in vivo by activated CD4+ T lymphocytes. AB - Donor CD8+ T lymphocytes injected into recipient mice incompatible at major histocompatibility complex (MHC) class I genes induce donor-specific CTL nonresponsiveness, attributed to the veto function of donor cells. Here we show that conditions leading to strong activation of CD4+ T cells, namely the presence in the recipient of foreign MHC class II determinants, lead to the apparent loss of veto function of donor cells. This "masking" of veto function is dependent on the dose of foreign MHC class II present. Veto function can be partially restored by treatment of recipients in vivo with CD4-specific antibody, a measure which has been shown to eliminate the function of CD4+ T cells in vivo. We conclude that CD4+ T cells activated by contact with antigen can interfere with the veto function of CD8+ T cells. Consequences of this finding are: (a) veto function of a sample cell population can be overlooked when activation of CD4+ T cells occurs simultaneously. (b) The balance between veto function of recipient cells and its abrogation might be responsible for the kind of graft-vs.-host reaction generated (CD8+ T cell-mediated and frequently lethal or CD4+ T cell-mediated and not lethal) when parental T cells are injected into recipients incompatible at MHC class I and class II genes. (c) A possible contribution of veto cells should be considered in several protocols in which donor hemopoetic cells were used in conjunction with CD4-specific antibodies to induce transplantation tolerance. (d) Veto function in vivo does not require a contribution of CD4+ T cells. PMID- 2567243 TI - Characterization of the subset of immature thymocytes which can undergo rapid in vitro differentiation. AB - Recently, we reported that thymocytes expressing the CD8 molecule on their surface can give rise to CD4+CD8+ double-positive and CD4+ single-positive progeny following intrathymic transfer into an irradiated host mouse. Thymcoytes expressing a high density of CD8, referred to as CD8hi, and those expressing a low density of the molecule, CD8lo, were both able to differentiate in vivo. In this study we examined the ability of these CD8+ thymocytes populations and of CD4-CD8- double-negative thymocytes to change their phenotype during brief in vitro culture. CD8+ thymocytes were prepared by anti-CD4 plus complement lysis followed by positive selection of the survivors on anti-CD8-coated plates. After 16 h of culture, greater than 60% of CD8+ thymocytes became double-positive. Both CD8hi and CD8lo cells were able to show this in vitro change: about 30% of the former and about 80% of the latter became double-positive. In contrast to this, double-negative thymocytes which had been depleted of cells expressing low densities of CD8 did not show such a phenotypic conversion in vitro. Further panning experiments suggested that all of the CD8+ thymocytes actually express a low surface density of the CD4 molecule which is undetectable in our cytofluorometric assays. PMID- 2567244 TI - Longitudinal study of leukocyte functions in homosexual men seroconverted for HIV: rapid and persistent loss of B cell function after HIV infection. AB - The early effects of infection with human immunodeficiency virus (HIV) were investigated in homosexual men who had seroconverted for anti-HIV antibodies. Leukocyte functional activities were determined in longitudinally collected peripheral blood mononuclear cell samples. During the first 10 months following seroconversion, anti-CD3 monoclonal antibody-induced T cell proliferation, monocyte accessory function and T helper activity on B cell differentiation in a pokeweed mitogen-driven system were not affected. In contrast, from the moment of seroconversion on, B cells of seroconverted men failed to produce immunoglobulin in the pokeweed mitogen-driven system. This defect was not restored by addition of normal CD4+ T cells. Immunoglobulin synthesis induced by Staphylococcus aureus and interleukin 2 decreased gradually, until it was completely lost 10 months after seroconversion. In addition, proliferation in response to anti-IgM or Staphylococcus aureus by B cells from HIV seroconverted men was decreased. The lack of inducible in vitro B cell activity was not accompanied by elevated spontaneous Ig synthesis by B cells of the seroconverted men. In the second group of men studied during the 2nd year following seroconversion, T helper activity on normal B cell differentiation significantly decreased, whereas anti-CD3-induced T cell proliferation and monocyte accessory function were not significantly affected. Our results demonstrate that in almost all HIV-infected individuals B cell functional defects are the first leukocyte abnormalities observed preceding defects in T helper activity. PMID- 2567245 TI - T lymphocyte heterogeneity in old and young mice: functional defects in T cells selected for poor calcium signal generation. AB - An increase in cytoplasmic free calcium ion concentration is thought to play a critical role in the entry of resting T lymphocytes into the mitotic cycle. Not all murine T cells, however, generate such a calcium signal when exposed to mitogenic doses of concanavalin A (Con A), and the proportion of nonresponsive cells increases with age in adult mice. Since the frequency of T cells able to generate cytotoxic or lymphokine-secreting cells in culture also declines in old age, we have speculated that the defect in changes in Ca2+ concentration might underlie this functional deficit. To examine the relationship between functional competence and the calcium signaling pathway, we performed limiting dilution analyses of T cells enriched (by a fluorescence-based cell-sorting method) for high or low calcium signal generation. We found that rapid Ca2+ signal generation after Con A exposure did indeed correlate well with functional competence in Con A-stimulated limiting dilution cultures. Furthermore, selection for Ca2+ signals induced by anti-CD3 antibody, or even by ionomycin, similarly enriched for the functionally competent set of T cells. The ionomycin result, in particular, suggests that the difference between the reactive and nonreactive sets of T cells is unlikely to result simply from alterations in receptor-linked signal transduction pathways. We discuss a model in which immune senescence involves an age-dependent increase in a subset of T cells in which changes in cytoplasmic Ca2+ concentration are relatively difficult to induce by either receptor dependent mitogens or ionophores. PMID- 2567246 TI - Functional characterization of the CD45R (2H4) molecule on CD8 (T8) cells in the autologous mixed lymphocyte reaction system. AB - In the present study, we have investigated the molecular basis for the immunoregulatory function of CD8 cells after autologous mixed lymphocyte reaction (AMLR) activation. We demonstrated that the CD8+CD45R+, but not the CD8+CD45R- subset of cells effected suppression following AMLR activation. In contrast, cytotoxic activity against alloantigens resided in both the CD8+CD45R+ and CD8+CD45R- subsets of cells. Biochemical analysis showed that on CD8 cells, the 220-kDa isoform of the LCA/T200 antigen family was better represented than the 200-kDa isoform, when compared to CD4 cells. The density of the CD45R antigen increased on CD8 cells following activation in AMLR and treatment of AMLR activated CD8 cells with either anti-CD45R antibody or anti-CD3 antibody abolished the suppressor function of these cells. In contrast, treatment of AMLR activated CD4 cells with anti-CD45R, but not anti-CD3 antibody, abolished the suppressor/inducer function of these cells. The results suggest that the CD45R antigen as well as CD3 T cell receptor complex have an important role in the suppressor function of AMLR-activated CD8 cells. PMID- 2567247 TI - In vivo depletion of BoT4 (CD4) and of non-T4/T8 lymphocyte subsets in cattle with monoclonal antibodies. AB - The effect on certain immune responses of depleting two distinct lymphocyte subpopulations in vivo by inoculating calves with monoclonal antibodies (mAb) was examined. An mAb directed against the BoT4 antigen (the bovine homologue of CD4) effectively removed the BoT4+ lymphocytes from peripheral blood mononuclear cells (PBM). Compared to controls, treated calves showed a reduced antibody response to human O red blood cells and to ovalbumin. PBM prepared from BoT4-depleted animals also had a significantly reduced ability to respond in vitro to the mitogens phytohemagglutinin, concanavalin A and pokeweed mitogen. An mAb directed against a second numerically large bovine lymphocyte subpopulation i.e. BoT2-, BoT4-, BoT8- (CD2-, CD4-, CD8-), that may be homologous to the CD4-, CD8- cells in man and rodents that synthesize the gamma/delta+ T cell receptor, was also used for in vivo depletion. Compared to controls, calves depleted of this subpopulation showed an enhanced antibody response. The proliferative response of PBM to pokeweed mitogen was also significantly increased but responses to concanavalin A and phytohemagglutinin remained unchanged. The results suggest this lymphocyte subpopulation has a nonspecific suppressor activity acting on B cell responses either directly or through an effect on T helper cells. The non-T4/T8 cells are found extensively in the epithelium and lamina propria of the mucosa of the alimentary tract but not in T cell areas of the lymph nodes, tonsil and spleen. These non-T4/T8 cells may thus be, or contain, an intraepithelial lymphocyte population with a suppressor function. PMID- 2567248 TI - Peripheral blood stem cells collected before and after leukapheresis in the very early remission phase of hematopoietic malignancies. AB - Human peripheral blood obtained after chemotherapy-induced remission in hemopoietic malignancies has been suggested to be a potential substitute for autologous bone marrow as regards autologous hematopoietic reconstitution. The schedule and consequences of early leukapheresis are, however, still imprecise. We report a study performed in two series of, respectively, 10 and 14 patients where sequential leukapheresis (total number = 84) was evaluated with regard to colony-forming unit (CFU) potency. Our data demonstrate that adequate numbers of progenitor cells can be collected by leukapheresis and that, even when this is performed at an early stage after remission, subsequent hematopoietic reconstitution is not impaired. PMID- 2567249 TI - Marrow aplasia developing 3 years after treatment with busulphan for chronic myeloid leukaemia. AB - A 31-year-old woman with Philadelphia chromosome-positive chronic myeloid leukaemia (CML) was treated intermittently with high-dose busulphan over a 6-yr period (total dose 1320 mg). 3 yr later (after receiving no further cytotoxic drugs) she developed pancytopenia and marrow aplasia of relatively abrupt onset. Transfusion of reconstituted blood-derived stem cells (collected 7 yr previously) re-established chronic phase CML. These events are more consistent with 'stem cell exhaustion' than with an acquired marrow microenvironmental defect occurring in the course of CML. The contribution of busulphan is uncertain. PMID- 2567250 TI - The developmental expression pattern of a new murine homeo box gene: Hox-2.5. AB - To examine the possible role of homeo box genes in murine development we have studied the structure and expression pattern of Hox-2.5, a newly isolated homeo box gene that maps to the left end of the mouse Hox-2 locus on chromosome 11. The sequence of the Hox-2.5 homeo box has been determined. It is highly homologous to Hox-1.7 and Hox-3.2, demonstrating extended conservation among three homeo box complexes in the mouse. Northern and in situ hybridization analyses of Hox-2.5 demonstrate a novel, regionally restricted pattern of expression in developing mesoderm and neurectoderm. We detect localized Hox-2.5 transcripts as early as 8.5 days postcoitum. The expression pattern of Hox-2.5 was analyzed over the next 3 days of ontogeny, as well as in later embryonic, newborn, and adult stages. Three-dimensional reconstruction of Hox-2.5 transcript localization within the central nervous system of early embryos clearly illustrates the neural expression domain. Although the Hox-2.5 expression pattern is regionally restricted during all of these stages of development, the pattern changes along the anteroposterior and dorsoventral axes of the CNS as the embryo undergoes complex morphogenetic movements and cytodifferentiation. PMID- 2567251 TI - The Drosophila fsh locus, a maternal effect homeotic gene, encodes apparent membrane proteins. AB - The maternal effect gene fsh is involved in the establishment of segments and the specification of their identities; the progeny of mutant females are missing portions of thoracic and abdominal segments, and may have homeotic transformations of third thoracic segments to second thoracic segments. The fsh locus interacts synergistically with loci such as Ubx and trx in the production of homeotic transformations. We have characterized cDNA clones corresponding to the major fsh transcripts expressed in ovaries and early embryos, and to a pupal transcript. The expression of fsh transcripts in ovaries is restricted to the germline; in developing embryos, transcripts are found throughout the cytoplasm. The different ovarian/embryonic transcripts (7.6 and 5.9 kb) are generated by use of alternative polyadenylation and splice sites. These transcripts encode two large predicted proteins of 110 and 205 kDa that have unusual amino acid compositions: 40% of the residues are glycine, alanine, or serine, and there are several regions of homopolymers and simple sequence repeats. Hydropathy analysis indicates that these proteins span the membrane. We suggest that the expression of fsh proteins in the membrane of the embryo is required for proper functioning of genes such as Ubx in the specification of segmental identity. PMID- 2567252 TI - Association and sibpair analysis for the HLA, Gm, Km, and insulin polymorphisms in multiplex IDDM families. AB - A log-linear model was used to analyze three-way interactions between IDDM and pairs of genetic markers. To do this, a special sample dataset was selected by taking one affected and one unaffected child from each family. Some three-way interactions were found for associations between insulin-dependent diabetes mellitus (IDDM), HLA-DR, and DQ restriction enzyme fragment length polymorphism (RFLP) patterns. No three-way interaction was found for IDDM, HLA-DR, and Gm or Km. An extended sibpair analysis was applied to the HLA-B,DR loci and to the Gm, Km, and insulin gene polymorphisms. The well-known result for IDDM and HLA was reproduced. For Gm, Km, and the insulin gene no cosegregation with IDDM could be found. PMID- 2567253 TI - HLA DR4-DQw3.1 and 3.2 haplotypes among insulin-dependent diabetics and their unaffected sibs in the GAW5 data. AB - Almost all human leukocyte antigen (HLA) haplotypes positive for HLA-DR4 also carry the DQw3 specificity, which appears in one of two allelic forms, DQw3.1 or DQw3.2. Previous studies have shown that the frequency of the HLA DR4-DQw3.2 allele is approximately 95% among DR4-positive haplotypes of insulin-dependent diabetics (IDDM), but only 70% in DR4-positive haplotypes of unaffected individuals. Because this difference could be due to ethnic heterogeneity, it is important to establish whether the frequency of the DQw3.2 allele is also increased when haplotypes of diabetics are compared to those of "matched" unaffected individuals, as can be done within families. We have used the Genetic Analysis Workshop 5 (GAW5) data for this purpose. In every family, each parental DR4-bearing haplotype was categorized as "IDDM" if it appeared in any affected parent or offspring, or as "control" if not. When this was done, the frequencies of the DQw3.2 and 3.1 allele in 80 IDDM haplotypes were 94% and 6% respectively but 67% and 33% in 15 control haplotypes. This difference between the two kinds of haplotypes is highly significant (P less than 0.005). PMID- 2567254 TI - Clues to IDDM pathogenesis from genetic and serological traits in multiply affected families. AB - A scheme is outlined for analyzing the genotypic contributions of two unlinked loci in producing a disease, using DR and the 5' insulin locus (INS) in insulin dependent diabetes mellitus (IDDM) as examples. Although genotypes of both DR and INS play roles in IDDM susceptibility, both the relatively small size of the Genetic Analysis Workshop 5 (GAW5) data set and the apparently limited magnitudes of the contributory effects prevent the identification of the exact nature of the association of these two loci in disease causation. The Gm allotypes showed no association with IDDM, either alone or in combination with other variables. Association of reactivity among the six strains of Coxsackie B virus is described, with no evidence of associations with DR type and IDDM found. The unaffected offspring segregated DR alleles according to expectations, while the segregation of affected alleles revealed the various contributions of DR alleles to IDDM pathogenesis, with the suggestion that DR4 from fathers is more diabetogenic than that from mothers. Lastly, a method is described for revealing the accuracy of typing in family data, and applied to RFLP variants subdividing DR3. PMID- 2567255 TI - HLA DQ beta 3.2 identifies subtypes of DR4+ haplotypes permissive for IDDM. AB - The HLA class II-related susceptibility to type I insulin-dependent diabetes mellitus (IDDM) is examined in 94 multiplex families sorted by the presence or absence of a DR4+ haplotype in at least one diabetic family member. The families with DR4+ haplotypes are then sorted by the presence or absence of a DR4-linked DQ beta 3.2 allele. Further analysis assumes each multiplex family to represent a single diabetic genetic event and identifies the HLA class II haplotype(s) present in all affected members. The DQ beta 3.2 allele is present in over 95% of the multiplex families where DR4+ haplotypes segregate with IDDM, implying a major permissive role in determining susceptibility to IDDM. PMID- 2567256 TI - Genetic analysis of IDDM: the GAW5 multiplex family dataset. AB - In a collaborative effort by 12 centers from Europe and North America, data were assembled from 94 multiplex families with insulin-dependent diabetes mellitus (IDDM) for analysis of genetic and other factors of possible etiological importance. The dataset contains information on the following genetic markers: HLA-DR beta and -DQ beta restriction fragment length polymorphisms (RFLPs), three RFLPs detected with two probes that map 5' to the insulin gene, the serologically defined HLA loci, and the immunoglobulin allotypes. Data also were included for auto-antibodies to insulin and pancreatic islet cells as possible indicators of pathogenesis and for antibodies to certain viruses that have been implicated as "triggering" agents in IDDM. Medical history of family members was obtained by means of a uniform questionnaire. Identical copies of the dataset were distributed to anyone wishing to participate in the analysis for the IDDM component of GAW5. The multiplex IDDM family dataset is now available on request for further analysis. PMID- 2567257 TI - HLA and insulin gene associations with IDDM. AB - The HLA DR genotype frequencies in insulin-dependent diabetes mellitus (IDDM) patients and the frequencies of DR alleles transmitted from affected parent to affected child both indicate that the DR3-associated predisposition is more "recessive" and the DR4-associated predisposition more "dominant" in inheritance after allowing for the DR3/DR4 synergistic effect. B locus distributions on patient haplotypes indicate that only subsets of both DR3 and DR4 are predisposing. Heterogeneity is detected for both the DR3 and DR4 predisposing haplotypes based on DR genotypic class. With appropriate use of the family structure of the data a control population of "unaffected" alleles can be defined. Application of this method confirms the predisposing effect associated with the class 1 allele of the polymorphic region 5' to the insulin gene. PMID- 2567258 TI - Linkage data on affective disorders in an epidemiologic context. AB - Recent data on chromosomal linkage markers in affective disorders indicate that single locus inheritance is involved in these disorders. The presence of a single locus has not been detectable using segregation analysis on family study data from large populations, possibly because of both heterogeneity and birth cohort and sex differences in diagnostic frequencies. PMID- 2567259 TI - Restriction fragment polymorphisms of the HLA-DR, HLA-DQ, and insulin gene regions in IDDM: the GAW5 data. AB - The primary aim of the insulin-dependent diabetes mellitus (IDDM) component of Genetic Analysis Workshop 5 (GAW5) was to collect and analyze new data on DNA polymorphisms closely linked to the HLA-D region and the insulin gene. The probes and restriction enzymes described here were used by all ten participating labs, and the data from Southern blotting were interpreted and reported according to conventions developed for the Workshop. These DNA data on members of 94 families with two or more IDDM sibs constitute the largest such sample available. The data were used in most of the analyses presented at the Workshop meeting, and are available on request. PMID- 2567260 TI - The insulin gene and susceptibility to IDDM. AB - The association between insulin-dependent diabetes mellitus (IDDM) and an allele of a restriction fragment length polymorphism (RFLP) 5' to the coding region of the insulin gene has raised the possibility that variation in the vicinity of the insulin gene confers susceptibility to IDDM. To test this hypothesis, the distribution of insulin gene sharing in affected sib pairs (ASPs) from the Genetic Analysis Workshop 5 (GAW5) families has been compared with that expected on the basis of random assortment. There is no deviation from random expectation in insulin gene sharing among 95 ASPs from families fully informative for the insulin gene. This is also true when insulin gene sharing is conditioned on HLA sharing, on the particular HLA DR types in ASPs, or on the parents' insulin allele classes. These results thus provide no evidence that variation at or near the insulin gene confers susceptibility to IDDM. However, we also used computer simulation to investigate how the insulin gene region could contribute susceptibility to IDDM without yielding evidence for distortion in insulin gene sharing in a sample comparable to that of GAW5. We found that various levels of insulin gene involvement in IDDM could generate a population association between the insulin gene RFLP and IDDM comparable to that reported in the literature, without producing significant distortion in insulin gene sharing of ASPs. PMID- 2567261 TI - Segregation analysis of two genetic markers in IDDM families under two-locus models. AB - The purpose of our study was to examine evidence for the role of a second genetic factor in the susceptibility to IDDM, in addition to that located in the HLA region. To do this, we have studied the joint segregation of HLA and another marker conditional on disease status using the IDDM families of GAW5, under a two locus model, one locus in the HLA region, the other close to the other marker. This marker has been, successively, Gm and the DNA polymorphism of the insulin gene 5' region. The study has been carried out using a segregation analysis method developed to make use of information on the segregation of two markers and the disease in nuclear families. The GAW5 data do not provide evidence for the role of a genetic factor in the Gm or insulin region in the etiology of IDDM. PMID- 2567262 TI - Linkage studies of HLA and insulin gene restriction fragment length polymorphisms in families with IDDM. AB - Linkage analysis of HLA DR antigen as well as DR and DQ restriction fragment length polymorphism (RFLP) data using the LIPED computer program and various three-allele disease locus models showed very close linkage to an insulin dependent diabetes mellitus (IDDM)-susceptibility locus. RFLP data alone were equal or superior to conventional HLA antigen typing in the linkage analysis. Insulin gene restriction fragment data were analyzed for evidence of either a susceptibility locus linked to the insulin gene or an effect of alleles at the insulin locus on the HLA-linked susceptibility gene. No evidence was found of any effect of the insulin gene, and it is suggested that alternative explanations of the reported population associations between the insulin gene and IDDM should be considered. PMID- 2567263 TI - Genetic variation in an inbred plant: variation in tissue cultures of soybean [Glycine max (L.) Merrill]. AB - Although soybean [Glycine max (L.) Merrill] grows as an inbreeding, generally homozygous, plant, the germplasm of the species contains large amounts of genetic variation. Analysis of soybean DNA has indicated that variation of RFLP (restriction fragment length polymorphism) markers within the species usually entails only two alleles at any one locus and that mixtures of such dimorphic loci account for virtually all of the restriction fragment variation seen in soybean (G. max), and in its ancestors, G. soja and G. gracilis. We report here that tissue cultures prepared from root tissue of individual soybean plants develop RFLP allelic differences at various loci. However, these newly generated alleles are almost always the same as ones previously found and characterized in other varieties of cultivated soybean (cultivars). This repeated generation of particular alleles suggests that much of the genetic variation seen in soybean could be the consequence of specific, relatively frequently employed, recombinational events. Such a mechanism would allow inbred cultivars to generate genetic variation (in the form of alternative alleles) in a controlled manner, perhaps in response to stress. PMID- 2567265 TI - Do H2 receptor antagonists have to be given at night? A study of the antisecretory profile of SKF 94482, a new H2 receptor antagonist which has a profound effect on daytime acidity. AB - Evening dosing has become standard for H2 receptor antagonists, because available agents inhibit nocturnal basal acid secretion more effectively than daytime stimulated secretion. We studied the optimal time of administration of a new high affinity long acting H2 receptor antagonist, SKF 94482, for the suppression of intragastric acidity using intragastric telemetry. Sixteen healthy subjects received SKF 94482 200 mg or placebo at 07:30, 17:30, and 21:30 h during four separate studies. Time (h) above pH 4 was (mean (SD] 1.1 (1.2) on placebo, 7.8 (5.0) on SKF 94482 given at 07:30, 5.75 (3.6) on SKF 94482 given at 17:30, and 6.1 (2.9) when given at 21:30. All treatment regimens were effective in increasing time above pH 4 (p less than 0.01). The efficacy of the morning dose of SKF 94482 indicates that the best time to give H2 receptor antagonists depends on their pharmacological properties. PMID- 2567264 TI - A comprehensive genetic map of murine chromosome 11 reveals extensive linkage conservation between mouse and human. AB - Interspecific backcross animals from a cross between C57BL/6J and Mus spretus mice were used to generate a comprehensive linkage map of mouse chromosome 11. The relative map positions of genes previously assigned to mouse chromosome 11 by somatic cell hybrid or genetic backcross analysis were determined (Erbb, Rel, 11 3, Csfgm, Trp53-1, Evi-2, Erba, Erbb-2, Csfg, Myhs, Cola-1, Myla, Hox-2 and Pkca). We also analyzed genes that we suspected would map to chromosome 11 by virtue of their location in human chromosomes and the known linkage homologies that exist between murine chromosome 11 and human chromosomes (Mpo, Ngfr, Pdgfr and Fms). Two of the latter genes, Mpo and Ngfr, mapped to mouse chromosome 11. Both genes also mapped to human chromosome 17, extending the degree of linkage conservation observed between human chromosome 17 and mouse chromosome 11. Pdgfr and Fms, which are closely linked to II-3 and Csfgm in humans on chromosome 5, mapped to mouse chromosome 18 rather than mouse chromosome 11, thereby defining yet another conserved linkage group between human and mouse chromosomes. The mouse chromosome 11 linkage map generated in these studies substantially extends the framework for identifying homologous genes in the mouse that are involved in human disease, for elucidating the genes responsible for several mouse mutations, and for gaining insights into chromosome evolution and genome organization. PMID- 2567267 TI - [Effects of nizatidine, a new histamine H2-receptor antagonist, on gastric acid secretion and various gastric and duodenal lesions in rats: comparison with cimetidine]. AB - We examined the antisecretory and antilesion activities of nizatidine in rats. Male SD or Donryu rats (200-260 g) were used under fasted or fed conditions. Nizatidine, given orally or parenterally (intraperitoneally, subcutaneously or intraduodenally) at 0.3-150 mg/kg, inhibited both basal (pylorus-ligation preparations) and histamine-stimulated gastric acid secretion (acute fistula preparations) in a dose-dependent manner. The potency of nizatidine was 2 to 8 times greater than cimetidine when the ED50 values (mg/kg or mu mole/kg) of each agent were compared. The antisecretory activity of nizatidine, given orally, persisted for more than 3.5 hr, but disappeared 6 hr later. Nizatidine, given orally or subcutaneously at 0.3-150 mg/kg, prevented development of gastric lesions induced by water immersion, pylorus ligation (Shay), histamine, aspirin, or indomethacin in a dose-dependent manner. Duodenal ulcers induced by mepirizole were also markedly prevented with nizatidine. The potency of nizatidine on stress lesions or duodenal ulcers was about 20 or 14 times greater than that of cimetidine, respectively. Nizatidine, given orally 3 times a day for 4 weeks, significantly (P less than 0.05) accelerated the healing of acetic acid-induced gastric ulcers which were delayed by prolonged treatment with indomethacin. These results suggest that nizatidine is a useful drug for the treatment of peptic ulcers in man. PMID- 2567266 TI - Olsalazine or sulphasalazine in first attacks of ulcerative colitis? A double blind study. AB - Olsalazine (2 g/day) and sulphasalazine (3 g/day) were compared in a double blind three centre trial in 37 patients presenting with first attack of distal colitis. Sigmoidoscopic appearances, rectal biopsies, and symptom and stool diary records were used to assess benefit and adverse effects. Both groups showed a similar decrease in stool frequency (p less than 0.001). The proportion of unformed stools was also decreased, but to a lesser extent (p less than 0.05) in those taking olsalazine (78% v 55%; p less than 0.001) compared with those taking sulphasalazine (72% v 28%; p less than 0.001). There was a diminution in the proportion of stools containing blood in both groups (olsalazine: 61% v 22%; p less than 0.001/sulphasalazine: 67% v 37%; p less than 0.001). Sigmoidoscopic and histological appearances and clinical activity improved significantly and to a similar extent in both groups. Intolerance was encountered in two patients on olsalazine and four on sulphasalazine; intolerance to sulphasalazine being even higher (five of seven patients) in a preliminary study using a dose of sulphasalazine releasing the same amount of 5-aminosalicylic acid as 2 g olsalazine. Olsalazine was at least as effective as sulphasalazine in the treatment of new patients with distal colitis, and in a dose releasing an equivalent amount of 5-aminosalicylic acid was better tolerated. PMID- 2567268 TI - Surgical treatment of foot and ankle trauma: use of indirect reduction techniques. AB - A useful intraoperative technique to aid the surgical treatment of complex foot and ankle trauma is described. With mechanical distraction to help obtain bony reduction and improve exposure, it can be used in the acute or reconstructive situation. Indications and a step by step technique are outlined. PMID- 2567269 TI - Crush injuries of the foot with compartment syndrome: immediate one-stage management. AB - Severe crush injuries with compartment syndrome were treated in five patients by an immediate one-stage procedure. This procedure included the assessment of skin flap viability with accurate debridement of devascularized tissues. It was performed according to the split-thickness skin excision technique. Compartment pressures were measured and the fasciotomies were performed through open wounds or separate medial and lateral incisions. The medial incision was extended to release the tarsal tunnel. Fractures were reduced and internally fixed and exposed bones were covered with locally transposed muscles. Skin grafts, taken earlier for the skin viability assessment, were meshed and applied to replace skin loss. All wounds and fractures healed uneventfully with no major functional loss. In multiple trauma, the physician should maintain a high index of suspicion for early diagnosis and treatment of severe foot injuries. Early treatment leads to more desirable results, shorter hospitalization, and faster rehabilitation. PMID- 2567270 TI - [Augmentation of a frontoparietal bone defect with autogenous rib cartilage]. PMID- 2567271 TI - Mitochondrial DNA polymorphism in Finnish families with Leber's hereditary optic neuroretinopathy. AB - Leukocyte mitochondrial DNA (mtDNA) from 17 Finnish families with Leber's hereditary optic neuroretinopathy and 70 maternally unrelated controls as well as skeletal muscle mtDNA from four of the Leber families and three controls was analyzed with 30 restriction enzymes. By this means, over 10% of the nucleotides of mtDNA were screened. No major deletion or insertion was found in any of the mtDNAs studied. The restriction fragment patterns of mtDNA showed no evidence of mtDNA heteroplasmy (mixture of different mtDNA types) in either blood or muscle cells. In all, 24 mtDNA types were observed in the material. In the maternal lines of Leber families, 11 mtDNA types were found, indicating no recent common maternal ancestor for the Finnish Leber families. In spite of several previously unknown polymorphisms, no mutation of mtDNA could be found exclusively in families with Leber's disease. However, a couple of mutations leading to amino acid replacements of mitochondrially encoded proteins were observed in certain Leber families only. These mutations have occurred in genes coding for subunits of NADH dehydrogenase, suggesting that a defect of the respiratory chain complex I may cause Leber's disease. PMID- 2567272 TI - Isolation of a DNA probe of potential use for diagnosis of the fragile-X syndrome. AB - A new cloned DNA probe (U6.2), which recognizes polymorphisms near the locus for the fragile-X syndrome, was isolated. No recombinations were observed between the probe and the disease locus, although recombinations were observed with several other probes known to be located close to the fragile site. The locus defined by the probe, DXS304, cosegregated with the fragile-X phenotype in 29 informative meioses (zeta = 4.97, tau = 0.00). The degree of polymorphism at this locus and its proximity to the fragile-X locus makes it useful for carrier diagnosis and as a new starting point for attempts to clone the gene responsible for the disease. PMID- 2567273 TI - The human cystatin C gene (CST3), mutated in hereditary cystatin C amyloid angiopathy, is located on chromosome 20. AB - Hereditary cystatin C amyloid angiopathy has recently been shown to be caused by a point mutation in the cystatin C gene. To determine the chromosomal localization of the gene, 20 human-rodent somatic cell hybrids and a full-length cystatin C cDNA probe were used. Southern blot analysis of BamHI digested cell hybrid DNA revealed that the probe recognizes a 10.6 kb human specific fragment and that this fragment cosegregates with human chromosome 20. Therefore, the human cystatin C gene (CST3) was assigned to chromosome 20. PMID- 2567274 TI - Restriction analysis of chromosomal sequences homologous to single-copy fragments cloned from small polydisperse circular DNA (spcDNA). AB - Restriction fragments from the fraction of small polydisperse circular DNA (spcDNA) were cloned in pBR322. The spcDNA was prepared from cell cultures derived from an angiofibroma of a patient with tuberous sclerosis (TS). Such cultures have been shown previously to contain increased amounts of spcDNA. Four cloned spcDNA fragments containing single-copy sequences were chosen to characterize the homologous chromosomal DNA segments by restriction analysis. When used as hybridization probes, these four fragments generate well-defined nonvariable patterns in the chromosomal DNA from healthy donors. The restriction patterns obtained with one of the fragments (D-C4) can best be interpreted by assuming the presence of two copies of the homologous sequences in chromosomal DNA. A second sequence, A-B4, occurs at least 30-50 times in the haploid human genome. In both cases the duplicated regions span relatively large segments of DNA. PMID- 2567276 TI - A rapid method for detection of Y-chromosomal DNA from dried blood specimens by the polymerase chain reaction. AB - The alphoid satellite family is the only repetitive DNA family showing chromosome specificity. We have developed a simple, rapid, and reliable test for sex diagnosis based on detection of these sequences in undigested genomic DNA using the polymerase chain reaction. In our test, dried blood specimens were the source of DNA. When female DNA was used as a template for the reaction, only the expected 130-bp X-chromosome-specific fragment was detected, while with male DNA both the expected 170-bp Y-chromosome-specific and X-chromosome-specific fragments were detected. The Y-chromosome-specific fragment was further characterized by restriction enzyme analysis. The Y fragment was detectable when DNA obtained from an equivalent of 10 microliters of spotted blood was used in the reaction, whereas detection of the X fragment was possible with DNA from an equivalent of 5 microliters of blood. Our test may find various applications in newborn screening and in forensic science. PMID- 2567275 TI - Frequency of heterozygous complete hydatidiform moles, estimated by locus specific minisatellite and Y chromosome-specific probes. AB - Restriction fragment length polymorphisms identified with three locus-specific minisatellite probes and banding patterns with Y chromosome-specific probes have been examined in 39 cases of complete hydatidiform mole (CHM) and the parents. All 39 cases were shown to be androgenetic. Of the 39 cases, 8 were identified as heterozygous CHM using the minisatellite probes. Estimates for the total number of heterozygous CHM in the series ranged from 23%-29%, higher than previously reported. Of the eight identified heterozygous CHM, six had Y chromosome-specific sequences whereas two were female; this is not significantly different from the 2:1 ratio expected. The low frequency of 46,XX heterozygous CHM in the literature may reflect difficulties in distinguishing them from 46,XX homozygous CHM. Examination of RFLPs with a small panel of locus-specific minisatellite probes provides a powerful method of classifying hydatidiform mole, enabling the rare heterozygous 46,XX CHM to be accurately identified. PMID- 2567277 TI - The factor IX BamHI polymorphism: T-to-G transversion at the nucleotide sequence 561. The BamHI/MSPI haplotypes in blacks and Caucasians. AB - The polymerase chain reaction (PCR) method was used to amplify a 356-bp DNA segment containing the suspected BamHI polymorphic site of the factor IX gene. Following the enzyme digestions and gel electrophoresis, polymorphic genotypes ( ,+ and +/- types) were observed. The gene frequencies for the rare (+) allele are about 36% in blacks and 2% in Caucasians. The 356-bp DNA was further purified and sequenced. The sequencing gels revealed a single nucleotide substitution (T to G) at position -561 of the gene in blacks and Caucasians. The T-to-G transversion generated a new BamHI site (GGATCC,+type) from a nonenzymatic site, (TGATCC, type). PMID- 2567278 TI - Close linkage of MEN2A with RBP3 locus in Japanese kindreds. AB - The gene responsible for multiple endocrine neoplasia type 2A (MEN2A) has recently been assigned to the pericentromeric region of chromosome 10 in European Caucasian kindreds by linkage analysis using a DNA marker, interstitial retinol binding protein 3 (RBP3). We have found tight linkage between the MEN2A and RBP3 loci in Japanese MEN2A kindreds. The maximum lod score is 5.19 at a recombination fraction of 0.00. This result suggests that mutation of a certain gene close to RBP3 is responsible for MEN2A irrespective of ethnic backgrounds. PMID- 2567279 TI - Dicarboxylic aminoaciduria associated with mental retardation. AB - Five hundred mentally retarded children (of both sexes and under 15 years of age) referred to our institute were screened for aminoacid disorders. One case of dicarboxylic aminoaciduria was found in a girl. PMID- 2567280 TI - Renin locus restriction fragment length polymorphism. AB - The use of two genomic EcoRI fragments as probes is discussed. PMID- 2567281 TI - Frequencies and interactions of regulatory T cells. I. The balance between help and suppression regulates the primary immune response to keyhole limpet hemocyanin in vitro. AB - Frequencies and efficiencies of regulatory T cells from non-immunized mice were estimated in several assay systems differing from each other in cellular composition and antigen dose (NIP-KLH). The NIP-specific and total IgM responses were quantified. Using 10(4) syngeneic B cells and 50 micrograms/ml NIP-KLH, helper (Th) cells from 5 day immune donors were detected in frequencies of 1:3000 1:4000 in lymphnode and spleen T cells, with an efficiency of 70-90 ng IgM/Th cell in C57B1/6 mice. In non-immune spleen T cells, Th cells were observed in frequencies of 1:16,000-1:38,000, with comparable efficiency, but these Th cells appeared suppressed at increased T cell doses. Polyclonal activation led to the appearance of multiple independently regulated populations of Th cells with similar efficiencies. In the presence of 10(5) syngeneic spleen cells, treated once with anti-Thy-1 antibody and complement and 50 micrograms/ml NIP-KLH, suppressor activity was observed in the same T cell population. Similar to help, suppression fluctuated with increasing T cell numbers. Using 1 x 10(6) spleen cells and 50 micrograms/ml NIP-KLH as assay system, T cells enhanced the responses. Again, several independently regulated populations were observed, with efficiencies slightly higher than those of the above-described Th cells. By maintaining the cellular components of the assay system constant (10(4) B cells) and titrating the antigen, Th cell frequencies showed little variation up to 100 micrograms/ml NIP-KLH and were always suppressed at higher T cell numbers. At 200 micrograms/ml NIP-KLH, the frequency was increased to approximately 1:2000, and not suppressed, i.e., was identical to the frequency observed in mice immunized 5 days previously. Efficiencies increased with increasing doses of antigen. The results strongly indicate that regulatory T cell function shows "plasticity", in the sense that the appearance and the frequencies of helping and suppressing T cell populations highly depend on the micro-environment present in culture. PMID- 2567282 TI - Structural and evolutionary comparisons of four alleles of the mouse Igk-J locus which encodes immunoglobulin kappa light chain joining (J kappa) segments. AB - The Igk-J locus of the mouse encodes the immunoglobulin kappa light chain joining (J) segments. Four Igk-J alleles have been described on the basis of restriction enzyme length polymorphisms. The nucleotide sequences of the Igk-Ja allele (type strain, C.C58), Igk-Jc allele (type strain, SJL/J), and Igk-Jd allele (type strain, SK/CamRk) have been determined and are compared with the previously reported Igk-Jb allele sequence (type strain, BALB/c). The mouse sequences are also compared with published sequences for rat and human J kappa sequences. Far more differences were found between the Igk-Ja allele and the other mouse alleles than between any two of the latter. These result in two amino acid substitutions which distinguish the J2 and J3' segments of the Igk-Ja allele from the other three alleles. Use of the Phylogenetic Analysis Using Parsimony program to generate a phylogenetic tree strongly indicates that after divergence from the rat ancestor, there appears to have been an early split between the Igk-Ja allele and the evolutionary precursor of the other mouse alleles. There also appears to have been far less divergence from the ancestral condition in the Igk-Ja allele than in the other alleles. Also, the presence of only one convergent mutation among the four mouse alleles provides strong evidence against any crossing over within the Igk-J locus during the history of these alleles. Finally, the differences in rates of evolution of the Igk-J alleles are in marked contrast to the relatively uniform rates of divergence of four alleles of a mouse V kappa gene, Igk-VSer. PMID- 2567283 TI - Anti-idiotypic T cells in human schistosomiasis. AB - We have developed an in vitro model of granuloma formation for the purpose of studying the immunological components of granulomatous hypersensitivity in patients infected with S. mansoni. Our previous studies have shown that 1) granulomatous hypersensitivity can be studied by examining the cellular reactivity manifested as multiple cell layers surrounding antigen-conjugated polyacrylamide beads; and 2) this reactivity is a CD4 T cell dependent, macrophage dependent, B cell independent response which is antigenically specific for parasite egg antigens. We report here on idiotype - anti-idiotype receptor interactions involved in the regulation of granulomatous hypersensitivity in patients and former patients infected with S. mansoni. Five human monoclonal antibodies specific for parasite egg antigens were used to activate CD4 and CD8 T cell subsets. These activated anti-idiotypic T cells were then assayed for regulatory effector functions in an autologous in vitro granuloma model. PMID- 2567285 TI - The EcoRI RFLP of c-mos in patients with non-Hodgkin's lymphoma and acute lymphoblastic leukemia, compared to geriatric and non-geriatric controls. AB - We have used Southern blot analysis to type individuals for the presence or absence of a rare EcoRI RFLP at the c-mos proto-oncogene locus. This polymorphism has previously been reported to be associated with cancer. Ninety-eight patients with non-Hodgkin's lymphoma (NHL) or acute lymphoblastic leukemia (ALL) and 154 cancer-free individuals, including 108 geriatric patients with no family history of cancer, were studied. Because 4 geriatric patients (aged 67-94) were found to have the rate c-mos allele (A2), and the frequency of this A2 allele was no higher among the lymphoma/leukemia patients than among cancer-free individuals, it is unlikely that it constitutes a marker for NHL or ALL. PMID- 2567284 TI - Binding of Escherichia coli S fimbriae to cultured human endothelial cells. AB - The binding of Escherichia coli adhesins to human umbilical vein endothelial cells was studied by a cell monolayer enzyme-linked immunosorbent assay. S fimbriae displayed a concentration-dependent and saturable binding to the endothelial cells which was mediated by their sialylgalactoside-specific lectin activity. P fimbriae exhibited only low binding, and type 1 fimbriae exhibited no binding to these cells. PMID- 2567286 TI - Effects of dopamine receptor activation on the level of cyclic AMP in the trabecular meshwork. AB - We examined the effects of dopamine and of a selective DA1 agonist, fenoldopam, on the levels of cyclic AMP in the trabecular meshwork, freshly excised from porcine and canine eyes. As measured by radioimmunoassay, fenoldopam at a concentration of 10(-5) M caused a 4-fold increase in the cyclic AMP content, from a basal level of 24.4 +/- 1.9 to 101.8 +/- 6.3 pmol/mg protein, of the trabecular meshwork samples from porcine eyes. In tissue samples from canine eyes, fenoldopam at a concentration of 10(-4) M increased the endogenous cyclic AMP level from a basal value of 26.0 +/- 4.6 to 64.2 +/- 5.7 pmol/mg protein. Dopamine, although less potent, produced a similar response. Preincubation with a DA1 receptor antagonist, SCH 23390, inhibited the increase in cyclic AMP levels by 90%. Such inhibition did not occur with the alpha- and beta-receptor antagonists phenoxybenzamine and propranolol, respectively. This investigation demonstrates that adenylate cyclase-coupled DA1 receptors are present in porcine and canine trabecular meshwork tissue. PMID- 2567287 TI - Ornithine aminotransferase distribution in ocular tissues and retinas of cat and mouse. AB - Ornithine aminotransferase (OAT), a mitochondrial matrix enzyme, is genetically deficient in patients with gyrate atrophy of the choroid and retina. Histologically defined micro-samples (10 ng-6.8 micrograms dry weight) were dissected out from freeze-dried tissue sections of eyeballs of cat and mouse, and the OAT activities were determined by a newly developed microassay method in the ocular tissues and retinal layers. Very high specific activities of OAT, expressed on a dry weight basis, were found in the feline ocular tissues of ectodermal origin, that is, neuroretina, retinal pigment epithelium, ciliary processes and epithelium of iris. In cat and mouse retinas, high OAT activities were distributed in the inner retina with an activity peak in the inner plexiform and ganglion cell layers. Very low activity was present in the outer nuclear layer. The inner segments of photoreceptor cells, which are very rich in mitochondria, contained the highest OAT activity. In contrast, the outer segments of photoreceptor cells contained the low activity resulting from contamination by small pieces of inner segments. PMID- 2567288 TI - Haptoglobin DNA polymorphism in subterranean mole rats of the Spalax ehrenbergi superspecies in Israel. AB - We analyzed the genetic diversity and environmental correlates of the haptoglobin (Hp) gene by RFLP analysis of 121 subterranean mole rats, comprising 13 populations belonging to the 4 chromosomal species (2n = 52, 54, 58 and 60) of the actively speciating Spalax ehrenbergi superspecies in Israel. The following results were indicated: (i) Out of the six restriction enzymes we tested, we found polymorphism only in TaqI, with three allelic fragments, and described their geographic patterning. (ii) Genetic diversity of Hp was highest in the centre of the range. (iii) The two major alleles showed high significant and opposite correlation with humidity. (iv) Genetic differentiation analysis indicated that 40 per cent of the Hp variation was within, and 60 per cent between populations. We conclude that climatic selection plays an important role in haptoglobin genetic differentiation. PMID- 2567289 TI - Light- and electron-microscopical localization of calcitonin, calcitonin gene related peptide, somatostatin and C-terminal gastrin/cholecystokinin immunoreactivities in rat thyroid. AB - Parafollicular C cells of the rat thyroid contain several immunoreactive peptides including calcitonin (CT), calcitonin gene-related peptide (CGRP), somatostatin and a C-terminal gastrin/CCK immunoreactive epitope as shown at the light- and electron-microscopical levels. Adult thyroid C cells are strongly immunoreactive to CT and most of the cells also react strongly with CGRP antisera and weakly with a gastrin/CCK antiserum. The latter antiserum may cross-react with CGRP. This cross-reactivity probably only occurs at very high concentrations of CGRP observed in adult thyroid C cells, but not in intrathyroidal CGRP-containing nerves, nor in early neonatal C cells. In neonatal rats, somatostatin immunoreactive C cells are numerous and most of these cells are also CT and CGRP immunoreactive. In contrast, only few C cells display somatostatin immunoreactivity in adult rat thyroids. Sequential staining experiments revealed that some thyroidal C cells simultaneously express all four types of immunoreactivity. At the electron microscopical level, all of these immunoreactivities were observed in secretory granules of C cells. Double- and triple-staining experiments, moreover, documented that some peptides are co localized in the same granules. PMID- 2567291 TI - Serum alpha 1-antitrypsin deficiency associated with the common S-type (Glu264--- Val) mutation results from intracellular degradation of alpha 1-antitrypsin prior to secretion. AB - The S-type alpha 1-antitrypsin (alpha 1AT) deficiency allele differs from the normal M1(Val213) allele by a single amino acid substitution (Glu264----Val). To evaluate the molecular pathophysiology responsible for the reduced serum levels of alpha 1AT associated with the S-type allele, alpha 1AT gene expression was examined in blood monocytes, cells which normally produce alpha 1AT, as well as murine fibroblasts modified by retroviral gene transfer to express the S-type and normal M-type human alpha 1AT genes. Northern analysis and S1 protection analysis demonstrated that monocytes of M and S homozygotes both express 1.8-kilobase alpha 1AT mRNA transcripts in comparable levels and similar in structure. Pulse chase labeling studies demonstrated that both M and S monocytes synthesized and secreted a 52-kDa protein, but the S monocytes secreted significantly less. The cellular lysates of both M and S monocytes contained a newly synthesized 50-kDa precursor form of alpha 1AT, but the S monocytes contained reduced amounts. Pulse chase labeling in the presence of tunicamycin, an inhibitor of core oligosaccharide addition, demonstrated that S monocytes exhibited a selective inhibition of secretion of 45-kDa nonglycosylated alpha 1AT not observed in M monocytes. Consistent with these observations, murine fibroblasts modified by retroviral gene transfer to contain an integrated human S-type alpha 1AT cDNA demonstrated reduced secretion of alpha 1AT compared with fibroblasts containing an integrated human M-type alpha 1AT cDNA and also reproduced the abnormality of alpha 1AT biosynthesis observed with S-type monocytes. Furthermore, in the presence of leupeptin, an inhibitor of cellular proteinases, the S-type modified fibroblasts demonstrated a selective augmentation of human alpha 1AT secretion not observed for the M-type. Together, these observations are consistent with the concept that the single A----T mutation of the S-type alpha 1AT gene results in reduced cellular secretion of alpha 1AT because the newly synthesized S-type alpha 1AT protein is degraded intracellularly prior to secretion. PMID- 2567290 TI - Overexpression and purification of the recombinant Ca2+-binding protein, apoaequorin. AB - The small, monomeric Ca2+-binding photoprotein, aequorin, emits blue light by an intramolecular reaction when mixed with Ca2+. The photoprotein is made up of coelenterazine and molecular oxygen, bound noncovalently to apoaequorin (apoprotein). The chemical steps leading to light emission, involving the oxidative degradation of coelenterazine, have been studied extensively, but little is known about the active site and how the molecule catalyzes the oxidation of coelenterazine. The three-dimensional structure of the protein has not been determined and therefore answers to these questions have remained unavailable. The present paper describes a procedure for preparing fairly large amounts of apoaequorin and aequorin for X-ray crystallographic studies. It consists of fusing the apoaequorin cDNA to the signal peptide coding sequence of the outer membrane protein A of Escherichia coli, which is under the control of the lipoprotein promoter. When the cDNA was expressed in E. coli, a large excess of the recombinant protein was produced and released into the culture medium. Purification of the protein was accomplished by acid precipitation and DEAE cellulose chromatography. The procedure yielded 7.4 mg of recombinant apoaequorin with a purity greater than 95% from 200 ml of culture medium. On regeneration with coelenterazine, the recombinant aequorin was fully active with Ca2+. PMID- 2567292 TI - Prosomatostatin 1-64 is a major product of somatostatin gene expression in pancreas and gut. AB - Prosomatostatin (pro-SS) is a peptide of 92 amino acids which contains the extensively studied somatostatin (SS) 1-28 and SS 1-14 at the C terminus. Little is known about the N-terminal part of pro-SS. In previous studies, using a radioimmunoassay against pro-SS 20-36 (sequence deduced from human cDNA sequence) we have identified a peptide with a molecular mass of approximately 8000 daltons in extracts of pancreas and intestinal mucosa. Using a variety of chromatographic procedures we have now isolated this peptide from extracts of pancreas and intestinal mucosa from pigs. The isolated peptides were sequenced on an Applied Biosystems gas phase sequenator and cleaved with the Asp-N endopeptidase for sequencing of C-terminal fragments. The peptides had an amino acid sequence identical to human pro-SS 1-64. In effluent from isolated perfused preparations of porcine small intestine and pancreas we identified upon appropriate stimulation pro-SS 20-36 immunoreactive peptides that by isocratic high pressure liquid chromatography appeared identical to pro-SS 1-64. An identical peptide was identified in pig plasma. Thus, in pancreas and gut pro-SS processing gives rise to the same pro-SS 1-64 molecule in spite of differential processing of the C terminus (SS 1-14 in pancreas and SS 1-28 in gut). The eventual hormonal role of pro-SS 1-64 may now be evaluated. PMID- 2567293 TI - Muscle-specific regulation of a transfected rabbit myosin heavy chain beta gene promoter. AB - We have examined the transcriptional regulation of the rabbit myosin heavy chain (HC) beta gene by using DNA-mediated transfection experiments. To analyze the activity of the myosin HC beta promoter in a myogenic background, cultured myoblasts from 12-day-old chick embryonic breast muscle were transfected with a chimeric gene containing 781 base pairs of the promoter region fused to the gene for chloramphenicol acetyltransferase (CAT). As indicated by the transient expression of chloramphenicol acetyltransferase, the activity of the promoter in myoblast cultures increased at least 32-fold following differentiation and was selectively inhibited when myogenesis was blocked with 5-bromodeoxyuridine. Furthermore, RNase protection experiments showed that the in vivo myosin HC beta transcriptional initiation (or cap) site was utilized in the transfected skeletal muscle cells and also that the regulation of the exogenous promoter was similar to the induction of the endogenous skeletal alpha-actin gene. The results indicated that the exogenous promoter is regulated in a tissue- and stage specific manner. By creating progressive 5' deletions of the promoter, we showed that only the region extending -294 base pairs upstream from the cap site is necessary for the muscle-specific expression. Linker-scanner mutagenesis of this region indicated that the positive regulation in differentiated skeletal muscle is mediated by at least two distinct elements within the 5'-flanking region of the myosin HC beta gene. PMID- 2567294 TI - Unrecognized foreign body as a focus for delayed Serratia marcescens osteomyelitis and septic arthritis. Two case reports. PMID- 2567296 TI - False negative results with benzodiazepine screening test. PMID- 2567295 TI - Association of HLA antigen and restriction fragment length polymorphism of T cell receptor beta-chain gene with Graves' disease and Hashimoto's thyroiditis. AB - HLA antigen phenotypes and BglII restriction fragment length polymorphism of T cell receptor beta-chain (TCR beta) gene were analyzed in 61 patients with Graves' disease and 50 patients with Hashimoto's thyroiditis. The antigen frequency of HLA-Bw46 in both Graves' disease (23.0%) and Hashimoto's thyroiditis (24.0%) was significantly higher than that in normal population (8.0%), with relative risks (RR) of 3.45 [corrected P (Pc) less than 0.009] and 3.66 (Pc less than 0.02), respectively. Significantly increased frequency of HLA-B51 antigen was also found in Hashimoto's thyroiditis (40.0% vs. 16.3% in controls; RR, 3.42; Pc less than 0.002). Hybridization of BglII-digested DNA with TCR beta probe revealed two alleles of 9.3 and 8.6 kilobases. The allele frequency of 8.6 kilobases in Graves' disease (79%) and Hashimoto's thyroiditis (76%) was significantly higher (P less than 0.01 and P less than 0.05, respectively) than that in controls (64%). The frequency of homozygous state 8.6/8.6 was significantly increased in both Graves' disease (62%) and Hashimoto's thyroiditis (60%) over that in controls (39%); the RR of 8.6/8.6 in Graves' disease and Hashimoto's thyroiditis were 2.55 (P less than 0.01) and 2.31 (P less than 0.05), respectively. These results indicate that in Japanese subjects at least two loci are involved in the susceptibility to Graves' disease and Hashimoto's thyroiditis, one related to HLA and another to TCR beta. PMID- 2567297 TI - The gamma-glutamyltransferase/glutamine synthetase activity ratio. A powerful marker for the acinar origin of hepatocytes. AB - The activity of glutamine synthetase (GS) in hepatocytes isolated by digitonin collagenase perfusion from the perivenous region was more than 10-times higher than in cells isolated from the periportal region. This distribution was confirmed by immunohistochemical staining for GS of cells separated from either region. In contrast, in periportal hepatocytes, the activity of gamma glutamyltransferase (GGT) was 3-4 times as high as in perivenous hepatocytes. This acinar distribution was also confirmed histochemically. The striking reciprocal acinar distribution of these two enzymes, now observed by direct biochemical analysis of selectively isolated hepatocytes, confirms the earlier qualitative differences observed by histochemistry and immunohistochemistry. The GGT/GS ratio seems to serve as a powerful marker of the acinar origin of isolated hepatocyte populations. Preliminary data describing glutamine synthetase activity in plasma of some subjects with suspected liver dysfunction suggests this enzyme as a marker for pericentral damage. PMID- 2567298 TI - Alveolar echinococcosis of the liver. PMID- 2567299 TI - A monoclonal antibody against human kidney gamma-glutamyl transpeptidase: preparation, immunochemical, and immunohistochemical characterization. AB - To perform immunohistochemical study of the distribution of gamma-glutamyl transpeptidase in human organs, a highly specific antibody against the human enzyme is required. We prepared monoclonal antibody against gamma-glutamyl transpeptidase from human kidney, using the hybridoma technique. The antibody was of the IgG1 type and the light chain belonged to the kappa class. The antibody reacted specifically with the 63 KD heavy subunit of the enzyme. Examination of the specificity of the antibody performed by immunohistochemical staining of human kidney sections revealed that the antigen was localized on the brush border and along the basolateral membrane of the epithelial cells of both the convoluted and the straight portions of the proximal tubule. This antibody was also reactive in several human organs other than kidney, including epididymis, prostate, seminal vesicle, pancreas, and normal liver, and in human hepatoma. These findings indicate the existence of an antigenic determinant common to human kidney and other organs. The monoclonal antibody did not crossreact with mouse, rat, guinea pig, rabbit, or pig kidney. PMID- 2567300 TI - High-affinity GABA uptake in a subpopulation of somatostatin cells in rat pancreas. AB - The aim of this study was to localize the high-affinity uptake of [3H]-GABA in Langerhans islets of rats aged 2.5, 7.5, and 75 days. On high-resolution autoradiography, cells presenting characteristic somatostatin granules were labeled, whereas others containing similar granules appeared nearly devoid of silver grains. Immunogold detection with antisomatostatin antibodies and high resolution autoradiography suggested that uptake of GABA is indeed performed by somatostatin cells. To test the heterogeneity of uptake frequency in somatostatin cells, a second approach, coupling immunohistochemistry with anti-somatostatin, anti-PP, anti-glucagon, anti-glicentin, and anti-CCK antibodies, and low resolution autoradiography, was applied on paraffin sections. It demonstrated that the uptake ability is not characteristic of all the somatostatin cells but of only a subpopulation of them. A few cells not immunoreactive to the anti somatostatin antiserum also appeared to be able to take up GABA. Moreover, except for a rare few, the PP-glucagon-, glicentin-, and CCK-39-immunoreactive cells were not labeled by autoradiography. PMID- 2567301 TI - The multidrug transporter: rapid modulation of efflux activity monitored in single cells by the morphologic effects of vinblastine and daunomycin. AB - Double-label fluorescence microscopy was used to demonstrate the efflux activity of the multidrug transporter in single cultured cells. NIH3T3 cells expressing a transfected MDR1 gene (NIH3T3-MDR) were treated with vinblastine or daunomycin. The accumulation of vinblastine was monitored by examining the morphology of tubulin in cells, using immunofluorescence. Overnight treatment of drug-sensitive cells caused disassembly of microtubules and formation of paracrystals; the absence of vinblastine effects was evident by the presence of intact microtubules. Daunomycin accumulation was detected in nuclei using the inherent fluorescence of the drug with rhodamine epifluorescence microscopy. Drug efflux in multidrug-resistant cells was inhibited with verapamil. When multidrug resistant cells were treated overnight in vinblastine, an effect of 0.5 microM vinblastine on microtubules was seen only in the presence of verapamil. Similarly, when cells were treated with daunomycin, this drug accumulated in nuclei only when verapamil was present. When cells incubated with vinblastine and verapamil were washed free of drugs, they did not accumulate daunomycin in a subsequent incubation, indicating that the multidrug transporter was still active; this occurred even though the morphologic effects of vinblastine persisted. Cells incubated with vinblastine alone showed an immediate inhibition of efflux activity when verapamil was subsequently added with daunomycin. These results show that the efflux activity of the multidrug transporter can be rapidly manipulated by agents such as verapamil, despite a prior history of drug treatment, and that the effects of inhibition of the transporter are rapidly reversible. PMID- 2567302 TI - Bacterial contamination of dialysate in dialysis-associated endotoxaemia. AB - Bacteriological investigations and endotoxin (ET) determinations were performed during a routine haemodialysis session for six patients. The glucose free dialysate was prepared with untreated tap water. All patients were dialysed for 5 h. Pseudomonas aeruginosa was regularly isolated in numbers up to 10(7) cfu ml-1 from samples of the dialysate inflow, the dialysate site and the dialysate outflow. ET levels in the plasma of the patients increased continuously during haemodialysis and were always higher in the blood outflow line of the dialyzer than in the blood inflow. Despite the high bacterial counts in the dialysate and the increasing ET levels in the patients plasma neither bacteraemia nor fever was observed. The former is due to the impermeability of the dialyzer membrane for bacteria, the latter is explained by low pyrogenicity of P. aeruginosa endotoxin. Inspection of the dialyzer machines revealed that air-traps and heater-unit for the incoming (untreated) tap water before mixing with the dialysate concentrate were the only sites where high bacterial release was feasible, as this part of the machine escaped disinfection due to the construction of these devices. We recommend the regular disinfection of all parts of a dialyzer machine, including heating units, air traps and valves. PMID- 2567303 TI - Nasal carriage of Staphylococcus aureus treated with topical mupirocin (pseudomonic acid) in a children's hospital. AB - 2% mupirocin ointment applied intra-nasally for 5 days was assessed for elimination of nasal carriage of Staphylococcus aureus in 31 staff members in a children's hospital. Three volunteers failed to complete the trial because of side effects, i.e. buccal reddening and swelling, and unpleasant taste. During treatment staphylococcal nasal carriage was not found in any case; of the 24 post treatment nasal swabs taken 4 days after treatment 22 were still negative. Re colonization with S. aureus of different phage types occurred in the remaining two cases. PMID- 2567304 TI - Legionella spp. in a hospital hot water system: effect of control measures. AB - Potential sources of Legionella spp. in a university hospital were investigated over 3 years in order to gain better understanding of the ecology and transmission of this organism to hospitalized patients. The survey highlighted the contamination of the hot water system with high concentrations of legionellas (up to 10(6) cfu 1(-1]. Legionella pneumophila serogroup 6 was predominant followed by L. pneumophila serogroup 10. Serogroup 1 and other species (L. longbeachae, L. micdadei) were rarely isolated. Serogroup 6 was also the predominant cause of nosocomial legionellosis in 15 sporadic cases in immunocompromised patients from 1981 to 1987. In light of this problem, several control measures were tried consecutively. A disinfection cycle with 6 ppm free chlorine failed to eradicate legionellas because of difficulties with the plumbing system. Raising the temperature in hot water tanks to 80 degrees C was effective locally, but mixer tanks where cold and hot water (60-65 degrees C) are mingled in order to achieve 45 degrees C became the principal reservoirs. Disconnecting the mixer tanks, maintaining a temperature of 60 degrees C in the heating tanks and accelerating the flow rate in the hot water system proved to be the most useful measures. PMID- 2567305 TI - Plasmid analysis of 219 methicillin-resistant Staphylococcus aureus strains with uncommon profiles isolated in Lisbon. AB - During the years 1986 and 1987, 219 methicillin-resistant Staphylococcus aureus (MRSA) strains were isolated in a Lisbon hospital. Antimicrobial susceptibilities and genetic analysis showed that resistance to penicillin, methicillin, erythromycin (inducible phenotype), tetracycline, gentamicin, tobramycin, kanamycin, streptomycin, neomycin and trimethoprim were chromosomally encoded. Plasmid DNA was absent from 38.8% of the isolates. Constitutive erythromycin resistance was coded by three plasmids of c. 2.3 Md, c. 2.0 Md and c. 1.6 Md. Chloramphenicol resistance was mediated by two plasmids of c. 2.9 Md and c. 1.8 Md. Small cryptic plasmids of c. 1.65 Md, c. 1.2 Md and c. 1.0 Md were also detected. The majority of the strains revealed antigen 17, were lysed by phages 75, 89 and/or 85, and were either devoid of plasmid DNA, or possessed a c. 1.6 Md plasmid coding for constitutive erythromycin resistance or a c. 1.0 Md cryptic plasmid. These observations cannot rule out that the MRSA Lisbon isolates are a homogeneous group of strains that might have a common origin, and seem to be different from MRSA recently isolated in other countries. PMID- 2567306 TI - Changes in hygienic procedures reduce infection following caesarean section. AB - Infection following emergency Caesarean section was reduced significantly by changing certain hygienic procedures, i.e. cutting instead of shaving abdominal hair, reducing vaginal examinations prior to operation, swabbing the external genitalia with aqueous chlorhexidine gluconate, minimizing the traffic in the operating theatre and abandoning peroperative dilatation of the cervix. Wound infection was reduced from 5.6% to 5.1% and endometritis from 14.3% to 4.4%. PMID- 2567307 TI - A method to evaluate the cleaning and disinfectant action of surface disinfectants. AB - Surface disinfection tests, used to evaluate new disinfectants, do not take into account the effects of detergents or of the mechanical cleaning process. We describe methods which evaluate both the disinfection and cleaning effect of disinfectants on organic matter. When testing alcohols at high concentrations (greater than or equal to 70%) on blood spots contaminated with Staphylococcus aureus, we found that the organisms were trapped and fixed to the test surface, probably due to denaturation of the blood. This gave a low inactivating factor (IF), as well as a poor subjective cleaning effect (SC). If serum was used instead of blood, we observed less pronounced trapping, resulting in a high IF although the SC was still poor. When broth was used, both IF and SC were satisfactory. With alcohols at a concentration of 42%, trapping was markedly reduced which improved the SC in blood contamination, with serum or broth contamination trapping did not occur. However, 42% ethanol lost its killing effect (i.e. low IF), whereas 42% isopropanol still demonstrated a high IF. PMID- 2567308 TI - An outbreak of Trichuris trichiura in a mental handicap hospital. AB - An outbreak of Trichuris trichiura in a ward for the severely mentally handicapped affected 22 of 29 patients. There was concurrent carriage of non pathogenic Entamoeba histolytica in 59% of these patients. Heavy contamination of soil was found in the patients' recreation area. The management of the outbreak is discussed. PMID- 2567309 TI - A bacteriological survey of amputation wound sepsis. AB - A retrospective survey of 100 lower limb amputations performed for ischaemia were analysed to assess the influence of preoperative bacterial isolates and the use of prophylactic antibiotics on wound sepsis. Forty-eight per cent had previously undergone a vascular procedure to attempt limb salvage and 17% were diabetics. Benzylpenicillin was given preoperatively and continued for 5 days; diabetics received metronidazole in addition. A total of 51 isolates were obtained from 30 patients preoperatively; Staphylococcus aureus and Enterobacteriaceae each accounted for over 25%. Postoperatively, 74 isolates (20 multiple) were obtained with an overall sepsis rate of 40%. Those patients with a positive preoperative culture were significantly more likely to develop wound sepsis. There was no significant difference in wound sepsis rates for diabetics. In view of the range of organisms causing postoperative infection, we recommend prophylaxis with a broad spectrum antibiotic for amputations. PMID- 2567310 TI - Aeromonas species in aquaria: a reservoir of gastrointestinal infections? AB - To assess the role of aquaria in the epidemiology of aeromonas associated gastroenteritis, the prevalence and antibiotic susceptibility of aeromonads in ornamental aquaria were determined and compared to that of isolates obtained from patients with gastroenteritis. Thirty-eight (76%) of 50 aquaria, including those located in our hospital, yielded 61 isolates: 24 (39.3%) Aeromonas sorbria, 18 (29.0%) A. hydrophila, 15 (24.6%) A. caviae, and four (6.5%) which could not be speciated. In comparison, 41 (65%) of 63 faecal isolates were A. caviae. The aquarium isolates, in contrast to enteric isolates, were generally resistant to chloramphenicol, tetracycline, and trimethoprim/sulphamethoxazole. These findings indicate aquaria are unlikely sources of aeromonas-associated gastroenteritis, but their role as possible reservoirs for non-enteric infections with aeromonads needs to be defined. PMID- 2567311 TI - The milk kitchen, Sheffield Children's Hospital, before and after a review. AB - The milk kitchen at Sheffield Children's Hospital has recently been modernized to update equipment and practices. Microbiological assessments before, during and after this upgrading are described. Limits for viable counts in pasteurized feeds are suggested and the practices adopted to minimize the contamination of feeds are described. PMID- 2567312 TI - Atypical legionellosis: isolation of Legionella pneumophila serogroup 1 from a patient with aspiration pneumonia. AB - It has been suspected that the number of Legionella pneumophila infections is substantially higher than commonly reported, due to subclinical infections. We report a patient where the diagnosis of a legionella infection was made only because of a large-scale surveillance programme for legionellosis. The case demonstrates the possibility of legionellas being involved in lung disease though not necessarily causing overt legionellosis. PMID- 2567313 TI - Septic complications in relation to factors influencing the gastric microflora in patients undergoing gastric surgery. AB - Postoperative septic complications and micro-organisms found in primary infections were studied in 750 gastric operations performed between 1972 and 1986. The overall rate of primary infections was 23%. The infection rates were related to the diagnosis and to factors that could influence the colonization of the stomach. No significant differences in the rates of postoperative infections were found between patients who had received preoperative antibiotic prophylaxis and those who had not. In all groups of patients, aerobic and anaerobic gram positive and gram-negative bacteria and yeasts were isolated in primary infections. Enterobacteriaceae, enterococci and Bacteroides fragiis were more frequent in patients with gastric bleeding or carcinoma. PMID- 2567314 TI - Expenses incurred during a 5-week epidemic methicillin-resistant Staphylococcus aureus outbreak. PMID- 2567315 TI - Privatisation. PMID- 2567316 TI - A placebo-controlled trial of the effect of two preoperative baths or showers with chlorhexidine detergent on postoperative wound infection rates. PMID- 2567317 TI - The evolution of MHC restrictions in antigen recognition by T cells in a haploidentical bone marrow transplant recipient. AB - We have longitudinally followed the major histocompatibility complex (MHC) restrictions that govern the response of T lymphocytes to specific Ag in a child with severe combined immunodeficiency who was successfully transplanted by using T cell depleted haploidentical maternal bone marrow cells and immunized shortly afterwards with tetanus toxoid (TT) Ag. In the first year post-transplant, monocytes were of both donor and recipient origin whereas T and B cells were of donor origin. Three years after transplant, all monocytes and T and B cells were of donor origin. T lymphocytes taken from the child at that time and depleted in vitro of alloreactivity to paternal Ag proliferated in response to TT presented by maternal as well as paternal monocytes. A TT-specific T cell line established from these cells in the presence of maternal monocytes cooperated with maternal but not with paternal monocytes, whereas a TT-specific T cell line established in the presence of paternal monocytes cooperated with paternal but not with maternal monocytes and with monocytes derived from a paternal uncle who shared the haplotype inherited by the recipient from her father. These results show that long-term memory T cells restricted to recipient MHC Ag not shared with the bone marrow donor continue to circulate long after the disappearance of accessory cells of recipient origin. These T cells could potentially participate in a secondary immune response because they were shown to recognize TT presented by recipient fibroblasts induced to express class II MHC molecules following treatment with IFN-gamma. PMID- 2567318 TI - [Diagnostic and therapeutic applications of intracavernous injection of vasoactive drugs in impotence. Defense for the use of facilitating drugs. Part I- Pharmacology, classification and complications of active drugs]. AB - The article discusses drugs which promote erection when injected via the intracavernous (IC) route during consultation. The diagnostic and therapeutic applications in the treatment of impotence are discussed also. 25% of impotent patients noted an improvement after this treatment while 50% of patients suffering from impotence of psychological origin noted an improvement. Auto injection is also discussed. IC treatment now seems justified in most cases which have not responded to traditional therapeutic approaches and this includes cases of psychological origin. Vasoactive drugs can be described as being inducers (use of these drugs induces a rigid erection, even in the presence of the doctor), facilitating drugs (which produce a rigid erection only if sexual stimulation is present also) and inhibitors (which stop the erection). The former group (which has papaverine as leader) produces a significant number of side effects, not least of these being priapism; there is a risk of lasting iatrogenic impotence which is not negligible. These risks are reduced considerably when one uses facilitating drugs which, although less powerful, suffice in treating a large proportion of cases of impotence. Papaverine can not be replaced as a diagnostic drug but facilitating drugs should be used first in therapy and inducers should be used only if these facilitating drugs have failed. PMID- 2567319 TI - [Diagnostic and therapeutic applications of intracavernous injection of vasoactive drugs in impotence. Plea for the use of facilitator drugs. 2. Diagnostic and therapeutic applications]. PMID- 2567320 TI - Serotype-specific prevalence of Escherichia coli strains with EPEC adherence factor genes in infants with and without diarrhea in Sao Paulo, Brazil. AB - To examine interrelationships of classic enteropathogenic Escherichia coli (EPEC) serotypes, EPEC adherence factor (EAF) genes, and diarrheal disease, E. coli were studied from stools of 500 infants less than 1 y of age with acute diarrhea and 500 age-matched controls. EAF-containing (EAF+) E. coli of three common classic EPEC serotypes (O111:H-, odds ratio [OR] 36.0; O111:H2, OR 55.0; O119:H6, OR 3.7) were individually strongly associated with diarrhea, as were EAF+ strains of less common classic serotypes combined (OR 5.3). Among EPEC serogroups, neither EAF+ strains of nonclassic serotypes (OR 1.8) nor EAF-strains of classic (OR 2.2) or nonclassic (OR 1.4) serotypes were significantly associated with diarrhea. At least one EAF+ non-EPEC serogroup serotype (O88:H25) may represent an unrecognized EPEC serotype. Serotype-specific variation in the association of EAF+ E. coli with diarrhea suggests that other factors are also important in determining virulence; thus, both EAF detection and E. coli serotyping are desirable in studying the etiology of diarrheal disease. PMID- 2567321 TI - Production of diarrhea in the rabbit by a mutant of Escherichia coli (RDEC-1) that does not express adherence (AF/R1) pili. AB - Escherichia coli (RDEC-1) adheres to Peyer's patch and absorptive epithelium in the rabbit in the closely adhering manner characteristic of enteropathogenic and enterohemorrhagic E. coli. An adherence pilus (AF/R1) is important for adherence to Peyer's patch M cells in vivo and ileal brush borders in vitro. A nonpiliated mutant (42-2-37-8) of the RDEC-1 strain colonized the gut lumen less readily than the parent strain. The mutant adhered infrequently to Peyer's patch lymphoid follicle epithelium (22% vs. 84%, P less than .0001). Ileal close adherence was less frequent at 3 d, but by 9 d after inoculation had increased, approaching that of the parent RDEC-1 strain. However, adherence was focal, and fewer bacteria were present at each adherence site compared with the parent RDEC-1 strain. The result was a lower frequency of diarrhea (34% vs. 65%, P = .003) and mortality (9.4% vs. 27%, P = .035) with the 42-2-37-8 strain. Loss of AF/R1 pili compromised the ability of the RDEC-1 strain to adhere to Peyer's patch and absorptive epithelium and to produce diarrhea. PMID- 2567322 TI - [Preventive effect of urinastatin on cisplatin induced nephrotoxicity in rabbits]. AB - The effect of urinastatin (US) on the prevention of cis-diamminedichloroplatinum (CDDP) induced nephrotoxicity was examined in experiments on rabbits. They were divided into seven groups and treated intravenously as follows: Group 1; intravenous injection of CDDP 3mg/kg only, Group 2; 3% sodium chloride (15ml/kg/h, 1h) + CDDP, Group 3; 3% sodium chloride + CDDP + US (10,000U/kg), Group 4; Hydration A (350ml/body) + 3% sodium chloride + CDDP, Group 5; Hydration A + 3% sodium chloride + CDDP + US, Group 6; Hydration B (200ml/body) + 3% sodium chloride + CDDP, Group 7; Hydration B + 3% sodium chloride + CDDP + US. Creatinine clearance (Ccr) value, arylamidase (AA) activity in urine and gamma glutamyltranspeptidase (gamma-GTP) activity in urine were consecutively measured as indices of nephrotoxicity. Ccr values in groups 1-3 significantly decreased markedly whereas those in groups 4-7 did not. The change in AA activity and gamma GTP activity in urine suggests that no nephrotoxicity occurred in groups 5 and 7. Consequently, this experiment indicated that nephrotoxicity was prevented when CDDP was administered in 200mg/body hydration with both 3% sodium chloride and US. The recommended dose of US seems to be a 2,000U/kg intravenous injection before CDDP administration, followed by 8,000U/kg intravenous drip infusion for 2 hours after CDDP administration. PMID- 2567323 TI - Urinary excretion of renal brush-border enzymes in lepromatous leprosy--a preliminary investigation. AB - Activities of the brush-border enzymes, alkaline phosphatase, maltase, leucine aminopeptidase, and gamma-glutamyl transpeptidase, were measured in urine samples of 25 lepromatous leprosy patients and an equal number of age-matched healthy controls. None of the patients were shown to be suffering from any other systematic disease. The enzymatic activities were shown to be significantly elevated in leprosy patients when compared to controls. PMID- 2567324 TI - [A case of multiple endocrine neoplasia, type 1, with parathyroid adenoma, multinodular adrenal cortical hyperplasia, islet cell tumor and hyperplasia of thyroid with concomitant hypersecretion of gastrin and total glucagon-like immunoreactivity]. PMID- 2567326 TI - Inhibition of human lymphocyte stimulation by visible light and phthalocyanine photosensitization: mitogen and wavelength dependency. AB - Mitogenic stimulation of human peripheral blood lymphocytes is inhibited by phthalocyanine photosensitization using visible light. The mechanism of inhibition was studied using stimulating agents differing in their mode of action. Stimulation by the plant lectin phytohemagglutinin (PHA) was the least sensitive to inhibition by photosensitization, followed by the tumor promoter phorbol myristate acetate (PMA) and the calcium ionophore A23187. Mitogenic stimulation could also be inhibited by light only, but higher fluences were needed. The order of efficacy was blue greater than red greater than green light, and the response to A23187 was least affected, followed by an increased inhibition of the response to PHA and PMA stimulation. Possible targets and mechanisms for these effects are discussed. PMID- 2567325 TI - Photo-induced irreversible shortening and swelling of isolated cochlear outer hair cells. AB - Living outer hair cells were irradiated under conditions of fluorescent microscopy (epi-illumination through the objective) with UVR (waveband 340-380 nm), blue light (waveband 450-490 nm) or green light (waveband 515-560 nm) in the intracellular presence or absence of the fluorescent dyes fura-2 or 2',7'-bis (carboxyethyl) 5-(and 6-)carboxyfluorescein (BCECF). In response to UVR with or without intracellular fura-2 and to blue light in the presence of BCECF (irradiation intensities of 2-12 x 10(5) W/m2), the cells shortened and swelled within 15-30 s, accompanied by the formation of numerous cytoplasmic granulations. The cellular reactions were significantly delayed to 3 min by the addition of 1 mM of the radical scavengers p-phenylenediamine or n-propyl gallate. This protection suggests that free radicals, produced under UVR or under blue light irradiation in the presence of the sensitizer BCECF, are possible causative agents of this cell damage. The response of the photo-damaged cells, namely shortening and increase in volume, resembled the characteristics of hair cells exposed to an hypo-osmotic shock. This suggests that structural alterations of the cytoplasmic membrane and the sub-membrane cortex occurred under photo irradiation, and that these structures can be implicated in the maintenance of the elongated cylindrical shape of the outer hair cells, possibly by maintaining intracellular hyperosmolarity. PMID- 2567328 TI - Acid and alkaline phosphatase activity in bone following intense magnetic field irradiation of short duration. AB - Consideration of the biological effects of electromagnetic radiation is particularly important with regard to bone metabolism. Magnetic fields have been applied to the treatment of bone fractures, and similar fields are employed in the visualization of skeletal structures through magnetic resonance imaging (MRI). The present investigation addressed the effect of MRI-intensity (1 Tesla) magnetic fields on enzymatic indices of bone metabolism. Mice were exposed to intense static (direct current) magnetic fields for 30 min on each of 10 successive days. After this exposure regimen the primary spongiosum of the humerus was assayed for acid and alkaline phosphatase activity. No significant difference was detected in tissue levels of either enzyme in bones from control (n = 10), sham-exposed (n = 10), and magnetic field-exposed (n = 10) mice. These results suggested that the effects of magnetic field irradiation do not appear to involve modification of the activities of regulatory enzymes in bone. PMID- 2567327 TI - Studies of the UVC-sensitivity of non-tumorigenic and tumorigenic human cell hybrids (HeLa x skin fibroblasts). AB - The UVC-sensitivities of a non-tumorigenic and a tumorigenic human cell hybrid (HeLa x skin fibroblasts) are compared and contrasted. The tumorigenic cells differ from the non-tumorigenic cells in that they have lost one copy each of chromosomes 11 and 14. For exponentially growing cultures, the tumorigenic cells are considerably more resistant than the non-tumorigenic cells. For confluent cultures, the differential in photosensitivity is much less. Flow cytometric studies of cell cycle distributions of both exponentially growing and confluent cultures of these cells indicate that the differences in photosensitivity cannot be explained by differences in cell cycle distribution. Studies of the kinetics of potentially lethal damage repair (PLDR) in confluent cultures of both cell lines indicate little or no recovery over the first 6 h followed by an increase in survival over the next 12-24 h. These data are consistent with previously published observations in human skin fibroblasts where the kinetics of PLDR reflected the kinetics of thymine dimer loss. The data are not consistent with 6 4 photoproducts being a potentially lethal lesion since such damage is rapidly repaired in human cells. PMID- 2567329 TI - Role of the superoxide anion in the oxidative activation of the new antitumor drug BD40: a radiolysis study. AB - BD40, a new antitumor drug derived from 9-azaellipticine, is thought to have an oxygen-dependent metabolism in vivo. We have investigated the one-electron oxidation of this drug by gamma radiolysis using OH. free radicals as oxidants and the reaction of O2-. with the BD40 oxidized transient(s). The absorption spectrum of the one-electron oxidized free radical was determined by pulse radiolysis using OH. or N.3 as reactant. In the absence of O2 and O2-., the initial yield of disappearance of the drug is equal to 2.5 x 10(-7) mol J-1 independently of the initial concentration of the drug and of the dose rate. When BD40 is oxidized by OH. radicals in the presence of O2 and O2-., the yield is the same. This yield is halved if superoxide dismutase is present during irradiation. Superoxide anions do not react directly with the drug. Thus it is suggested that these radicals oxidize the BD40 free radical produced by oxidation with OH. Biological implications are discussed. PMID- 2567330 TI - Locus specificity for mutation induction in human cells exposed to accelerated heavy ions. AB - The relative efficiencies of two types of densely ionizing particles were compared for the induction of mutations at two distinct genetic loci in human cells. Mutations to 6-thioguanine resistance (hgprt locus) or to trifluorothymidine resistance (tk) locus were scored in TK6 human lymphoblastoid cells exposed to graded doses of 40Ar ions (470 MeV/amu, LET = 95-97 keV/microns) or 28Si ions (456 MeV/amu, 61 keV/microns). The autosomal tk locus was more efficiently mutated than the X-linked hgprt locus following heavy particle irradiations. This was predominantly due to the contribution of a class of slowly growing mutants scored at the tk locus. Silicon ions were more efficient per unit dose than argon ions for the induction of mutants at either locus. When the mutant yield for a particular ion was compared with particle fluence, similar numbers of hgprt mutants are induced by equal numbers of 40Ar or 28Si ions. Comparison of the number of tk mutants with particle fluence demonstrates an increased efficiency for 28Si ions over 40Ar. These data suggest that the LET-RBE relationship may be different for individual genetic loci in human cells. PMID- 2567331 TI - Interpretation of the increase in the frequency of neoplastic transformations observed for some ionising radiations at low dose rates. AB - The anomalous increase of transformation frequency with decreasing dose rate observed by Hill et al. (1982, 1984b) for mouse fibroblast cells irradiated with fission neutrons cannot be satisfactorily explained by current models of radiation action. Recently a new model has been proposed which predicts the enhancement of damage with prolongation of irradiation, for equal doses. This is applied to the transformation studies in an attempt to interpret the enhancement observed for some radiations but not for others. Evidence is presented which suggests that repaired double-strand breaks in the DNA of cells which survive are the precursors of transformation. A critical physical factor is the total irradiation time rather than the dose rate. Approximately 1 per cent of repaired surviving cells go on to transform. From the results an explanation emerges of why transformation enhancement at low dose rates is not observed for natural alpha radiation and for photons or electrons, but is observed for fission neutrons and fast iron ions. PMID- 2567332 TI - Early postirradiation chromatin degradation in thymocytes. AB - The decrease in the average DNA size in thymocytes starts soon after in vivo irradiation and at approximately 45 min reaches a plateau, thereafter showing only minor changes up to 3 h. This fall in extent of chromatin cleavage coincides with the accumulation of 1.0-1.5 kb DNA fragments. Double-strand breaks generated by endonucleases are not randomly distributed along DNA but clustered in such a way that they give rise to fragments of 1-5 nucleosomes in size. Cycloheximide treatment partially inhibits nuclease activity in nuclear extracts isolated from thymus of irradiated mice. This suggest that DNA fragmentation is an early event in programmed death of thymocytes mediated by irradiation. The data indicate that it requires protein synthesis and that it precedes release of polydeoxyribonucleotides. PMID- 2567333 TI - Radiation sensitivities are not related to the sizes of DNA supercoiled domains in L5178Y-R and L5178Y-S cells. AB - Survival of murine lymphoblasts L5178Y-R and L5178Y-S irradiated with 60Co gamma radiation was determined. The parameters of the survival curves were D0 = 1.18 Gy, n = 1.56 and D0 = 0.55 Gy, n = 1.00 for L5178Y-R and L5178Y-S cells respectively. The sizes of DNA supercoiled domains were estimated using sedimentation of nucleoids from cells irradiated with doses from 1 to 7 Gy. These sizes were 2.44 x 10(9) and 5.13 x 10(8) Da for L5178Y-R cells and 1.30 x 10(9) and 4.07 x 10(8) Da for L5178Y-S cells. Hence, higher radiosensitivity of L5178Y S cells was not compatible with the larger size of the DNA supercoiled domains, as suggested by Filippovich et al. (1982). We have not found any simple relation between the sizes of DNA supercoiled domains and the susceptibility of L5178Y sublines to ionizing radiation. PMID- 2567334 TI - Factors affecting and significance of G2 chromatin radiosensitivity in predisposition to cancer. AB - The frequencies of chromatid breaks and gaps in metaphase cells fixed 2 h after G2 phase X-irradiation (1 Gy) were in almost all cases at least two- to three fold higher in skin fibroblasts from individuals with genetic conditions predisposing to cancer than in comparable cells from clinically normal controls. Previously, we reported this response in all cancer-prone genetic disorders tested including ataxia telangiectasia, Bloom's syndrome, Fanconi's anemia, xeroderma pigmentosum (XP), familial polyposis, Gardner's syndrome, hereditary malignant melanoma, dysplastic nevus syndrome and cancer family members. One exception was XP-A. In this report we add information on skin fibroblasts from retinoblastoma, Wilms' tumor and XP-C patients, 13 clinically normal controls and six cell lines from fetal or infant cells. Factors affecting the response are identified and include pH, temperature, cell density, culture medium or serum, microbial contamination and visible light exposure (effective wavelength 405 nm). Because of experimental variability, known normal controls should be used in each group of assays. With adequate control of the above factors this response could provide the basis of a test for detecting individuals carrying genes that predispose to a high risk of cancer. PMID- 2567335 TI - Radiosensitization by quaternary salts of 5-nitroimidazole derivatives. AB - The radiosensitizing effects of five newly synthesized quaternary salts of 5 nitroimidazole derivatives on the survival of TC-SV40 mammalian cells have been measured. A toxicity study was carried out in order to determine the concentrations to be used in the radiosensitizing experiments. The oxygen enhancement ratio (OER) for TC-SV40 cells was 2.74. None of the five 5 nitroimidazole derivatives showed radiosensitizing activity in aerobic conditions, while in hypoxia their dose-modifying factors (DMF) at the concentration of 0.2 mmol dm-3 range from 1.52 to 1.03 in this order: unsubstituted pyridinium greater than carbamoyl pyridinium greater than trimethyl pyridinium greater than t-butyl pyridinium greater than imidazolium. This latter product at the concentration of 2 mmol dm-3 has a DMF of 1.64. As comparison, metronidazole was also tested on this cell line and its DMF at 0.2 mmol dm-3 was 1.35. The response-concentration dependences for the unsubstituted pyridinium 5 nitroimidazole derivative and for metronidazole (comparing charged and uncharged structures) showed the flattening response-concentration curve of quaternary compounds. The electron affinity was evaluated through the CNDO/S theoretical method, and an exponential relationship between these values and the DMFs of the pyridinium derivatives was demonstrated. PMID- 2567336 TI - Does the repair capacity of skin change with repeated exposure to X-rays? AB - A sensitive experimental design and data analysis were used to test rigorously whether the repair capacity in the skin of the mouse foot changes during a course of repeated 240 kVp X-ray doses. Any such changes might reflect saturation or induction of repair enzymes resulting from progressive radiation damage, but most importantly this assumption of equal effect per dose fraction is central to all analyses of multiple-fraction radiation treatments, and remained to be demonstrated conclusively in skin. An X-ray dose of 2.5 Gy was given two, eight, 14 or 20 times with an interfraction interval of 8 h. Individual skin reactions for each mouse were analysed separately, giving 139 estimates of the effectiveness of 2.5 Gy (approximately 35 in each of the four fractionation schedules). Regression analysis of effect per fraction versus number of fractions showed that there was no significant trend, i.e. the damage per fraction was constant regardless of the number of fractions used. The mean damage per fraction was 3.75 +/- 0.15 per cent (95 per cent CL) of the full underlying damage equivalent to transient moist desquamation, and the slope of this plot was 0.0075 per cent +/- 0.022 per cent (95 per cent CL) per fraction. It was concluded that the assumption of equal effect per fraction was not invalidated in mouse skin. Shorter interfraction intervals would not allow full repair between fractions, and this could be misinterpreted as a progressive loss of repair capacity in this type of experiment. This was tested in skin by giving 2.5 Gy doses two, eight and 14 times with a 1-h interfraction interval. Effect per fraction increased with number of fractions, by an extra 37 per cent from two to eight fractions and by a further 14 per cent from eight to 14 fractions, giving the illusion of loss of repair as predicted. This confirms the need to check that where loss of repair capacity is suspected, this is not due artifactually to incomplete repair between fractions in slowly repairing systems. PMID- 2567338 TI - Structural and biochemical evidence that scrapie-associated fibrils assemble in vivo. AB - Scrapie-associated fibrils (SAF) are a ubiquitous pathological feature of brains affected by scrapie and the other scrapie-like agents. They are composed of PrP, a heterogeneous glycoprotein which is also present in normal brain but not as SAF. The PrP protein associated with SAF is partially resistant to proteinase K, whereas the soluble form is not. It has been proposed that SAF do not exist as such in vivo, but rather self-assemble from subunit structures liberated from membranes by detergent extraction during purification. We have purified SAF by a method that does not employ proteinase K. We show that the PrP protein from infected but not uninfected brain is partially resistant to protease digestion before and after detergent extraction. Likewise, SAF can be sheared by sonication before or after detergent extraction. In addition, SAF from mice infected with different strains of scrapie have different sedimentation properties. Since SAF dependent properties exist before detergent extraction, then so must SAF. They are therefore not a detergent-induced artefact but most probably assemble in vivo. PMID- 2567337 TI - Dissection of the human CD2 intracellular domain. Identification of a segment required for signal transduction and interleukin 2 production. AB - To evaluate those residues in the 117 amino acids of the CD2 cytoplasmic domain required for transduction of T lymphocyte activation signals, a full-length human CD2 cDNA and a series of deletion and substitution mutants were inserted into the ovalbumin-specific, I-Ad-restricted murine T cell hybridoma 3DO54.8 using a retroviral system. The resulting cells express surface CD2 protein and unlike the parental murine line, are reactive with murine anti-human CD2 antibodies. Anti T11(2) plus anti-T11(3) antibody stimulation of cells expressing a full-length CD2 cDNA results in a characteristic rise in cytosolic-free calcium [( Ca2+]i), and subsequent IL-2 secretion that accompany CD2 stimulation in human T lymphocytes. Transfectants expressing CD2 delta C98 and CD2 delta C77, partially deleted CD2 molecules containing the entire extracellular and transmembrane CD2 segments but only 98 and 77 amino acids of the cytoplasmic domain, respectively, are also activated by anti-CD2 mAbs. In contrast, clones expressing more severely truncated CD2 structures, CD2 delta C43 and CD2 delta C18, are not stimulated. These data show that the cytoplasmic domain plays an essential role in transduction of activation signals via CD2, and that the segment between amino acid residues 253 and 278 is necessary for activation. This region contains two tandem repeats of the sequence PPPGHR, thought to form part of a putative cationic site. Disruption of the latter by site-directed mutagenesis does not affect IL-2 gene induction, suggesting that only one of the repeats is required for activating this function of the CD2 molecule. PMID- 2567339 TI - A tandem repeat sequence found in a heterogeneous fragment of UL of herpes simplex virus type 1. AB - We found a tandem repeat sequence in the region (designated BS7) in which restriction fragments BamHI D and SalI B overlap each other, near the centre of the unique long sequence (UL) of the herpes simplex virus type 1 (HSV-1) strain F genome. The SmaI physical map of BS7 was constructed, and the position of a heterogeneous SmaI subfragment from HSV-1 isolates and plaque-purified clones from a single strain was defined on the map. The maximum size difference in the SmaI subfragment was estimated to be 600 bp between these isolates and 100 bp between the clones. The 0.23 kb SmaI subfragment recloned from BS7 was sequenced, and was shown to contain a tandem repeat sequence consisting of 15 units of 12 bp, 5' TTGGGGCTGGGG 3'. These results suggest that the fragment length heterogeneity in the UL of HSV-1 isolates and clones is attributable to copy number variation of the tandem repeat sequence. PMID- 2567340 TI - Gene UL2 of herpes simplex virus type 1 encodes a uracil-DNA glycosylase. AB - An insertion mutant of herpes simplex virus type 1 has been constructed which carries the lacZ gene from Escherichia coli within the coding sequence of gene UL2, which is in the long unique region of the genome. In a one-step growth curve experiment this recombinant (called in 1601) grew as well as the wild-type (wt) parent virus, indicating that the UL2 gene is dispensable for growth in tissue culture. Analysis of in 1601 DNA with restriction endonucleases showed no detectable changes from the wt apart from the insertion. Extracts of cells infected with in 1601 possessed levels of viral DNA polymerase and alkaline exonuclease activities similar to those infected with the wt, but unlike the wt had negligible uracil-DNA glycosylase activity, suggesting strongly that the product of the UL2 gene is the uracil-DNA glycosylase. The sequence of the uracil DNA glycosylase gene of E. coli was recently published, and the encoded amino acid sequence of this shows clear similarity to that of UL2, confirming our results. PMID- 2567341 TI - Effect of analogues of S-adenosylmethionine on in vitro polyadenylation by vesicular stomatitis virus. AB - Other workers have reported that vesicular stomatitis virus makes aberrantly long polyadenylic acid [poly(A)] tracts in the presence of S-adenosylhomocysteine (S Ado-Hcy). In the work reported in this paper, the effects of various analogues of S-adenosylmethionine (S-Ado-Met) and ATP on polyadenylation in an in vitro transcription system were examined to determine whether S-Ado-Hcy exerted its effect on polyadenylation due to its relationship to S-Ado-Met or to ATP. It appeared that compounds which affected polyadenylation were those which were closely related to S-Ado-Met and that had the same L-aminoacyl side chain [(COOH) CH(NH)2-CH2-CH2-]; the nature of the substituent at the -S+(CH3)- position of S Ado-Met was less important. These analogues appeared to compete with S-Ado-Met for a binding site(s). These data support a model whereby compounds binding at an S-Ado-Met-binding site may have allosteric effects by causing or preventing conformational changes which are involved in polyadenylation reactions, perhaps by affecting the rate of polyadenylation or of termination. PMID- 2567342 TI - Synthesis of respiratory syncytial virus RNA in cell-free extracts. AB - A cell-free system has been used to synthesize respiratory syncytial virus (RSV) RNA in vitro. Polyadenylated species representing all size classes of RSV mRNAs were labelled. Some of the labelled RNA was seen in a CsCl gradient at the density characteristic of negative-strand viral nucleocapsids. Experiments using a thiotriphos-phorylated nucleotide indicate that RNA chains are initiated de novo in the cell-free system. PMID- 2567343 TI - Cross-reactive immunity among different serotypes of virus causing haemorrhagic fever with renal syndrome. AB - Spleen cells primed by Prospect Hill (PH) or Puumala (Pu) virus could cross-react with Hantaan virus (HV) 76-118 strain-infected target cells after in vitro stimulation with HV-infected cells, although anti-PH or anti-Pu immune serum showed no cross-reactive neutralizing (NT) activity to HV without complement. These results and our previous findings with cross-reactive cytotoxic T lymphocytes (CTLs) suggest that some epitopes recognized by CTLs might be common among the hantavirus genus, while the epitopes related to NT activity were mainly specific to each virus of this genus. Next, to evaluate the cross-reactive immunities demonstrated by in vitro study, we investigated the effect of transferring T lymphocytes and sera from BALB/c mice immunized with PH or Pu virus into nude mice before HV inoculation. Transferring T lymphocytes primed by PH or Pu virus reduced HV titres in lungs and spleens of nude mice, corresponding with the results of the in vitro CTL assays. Transferring anti-Pu immune serum also decreased HV titres in nude mice, which seemed to reflect complement dependent NT activity. Moreover ICR mice previously immunized with PH or Pu virus showed resistance to challenge with a lethal dose of the HV KHF strain, indicating that cross-reactive immunity induced by PH or Pu virus could protect ICR mice against pathogenic HV infection. PMID- 2567344 TI - Alpha 2-adrenoceptor antagonists potentiate the anticonflict and the rotarod impairing effects of benzodiazepines. AB - Putative interactions between the specific alpha 2-adrenoceptor antagonist idazoxan and benzodiazepines (BDZs) were examined in two different rat conflict models; Vogel's drinking conflict test (VT) and Montgomery's conflict test (MT) (the elevated +-maze). In the MT, idazoxan (0.031 mg/kg) produced anxiogenic-like effects, which were counteracted both by the triazolo-BDZ alprazolam (APZ; 0.2 mg/kg) and the conventional BDZ diazepam (DIZ; 0.2 mg/kg). In fact, the anxiolytic-like effects of APZ were significantly potentiated when co administering idazoxan. A tendency to such a phenomenon was seen also in rats treated with DIZ and idazoxan. In the VT, the anxiolytic-like effects both of APZ (1.0 mg/kg) and DIZ (4.0 mg/kg) were significantly enhanced when co-administering idazoxan (1.0 mg/kg) in a dose not affecting the behavior per se. Similar potentiating phenomena by behaviorally inert doses of alpha 2-adrenoceptor antagonists (idazoxan 1.0 mg/kg; yohimbine 2.0 mg/kg) were seen with regard to the ataxic/sedative effects of the BDZs (APZ 0.25 mg/kg; DIZ 1.5 mg/kg). The present results provide further support for the notion that the anxiolytic-like effects of BDZs are not related to attenuation of Locus Coeruleus activity. In addition, it is suggested that the potentiation caused by the alpha 2 adrenoceptor antagonist is mediated via a noradrenaline induced increase in signal-to-noise ratio in target neurons of the brain noradrenergic system. PMID- 2567345 TI - Selective D-1 dopamine receptor agonist effects in hyperkinetic extrapyramidal disorders. AB - The motor and cognitive effects of a selective D-1 dopamine receptor agonist, SKF 39393, were assessed in patients with Huntington's disease, Gilles de la Tourette's syndrome, tardive dyskinesia, and torsion dystonia, using a double blind placebo-controlled design. Over daily doses ranging from 3.2 to 32 mg/kg and treatment intervals extending from one to seven weeks, no consistent changes could be discerned. The contribution of D-1 receptor mediated mechanisms to the pathophysiology of hyperkinetic extrapyramidal disorders remains uncertain. PMID- 2567346 TI - The use of H2-receptor antagonist in the treatment of peptic ulcer disease in children. AB - We report our 20 years' experience on the management of 78 children with primary chronic peptic ulcer, giving special emphasis to the recent 32 patients (29 duodenal ulcer, 3 gastric ulcer) who received H2-antagonist therapy guided by endoscopic surveillance. The most striking observation was the marked decrease in the incidence of "failed medical treatment necessitating surgery" from 34.8% (16/46) in the pre-H2-antagonist era to 3.1% (1/32) in the H2-antagonist era. H2 antagonist was uniformly well tolerated and without side-effects. In 29 patients with duodenal ulcer, a 6-week therapeutic course resulted in complete ulcer healing in 22 (75.9%). Further therapeutic courses resulted in complete ulcer healing in four additional patients (13.8%). There was only partial ulcer healing in two patients (6.9%) and nonhealing in one patient (3.49%). There were 11 episodes of recurrence in nine patients (symptomatic 5, asymptomatic 6). Recurrence was more frequent when nocturnal-dose maintenance was not given (8/11 recurrences). All recurrences responded satisfactorily to further H2-antagonist therapy. Patients with gastric ulcer responded even more favorably, all three healing completely with a single therapeutic course and not requiring maintenance therapy. We conclude that H2-antagonist is safe and highly effective for healing peptic ulcer in children. For duodenal ulcer, maintenance therapy ensures a satisfactory remission rate. PMID- 2567347 TI - Multi-beat clonus in a patient without an upper motor neuron lesion. A case report. AB - The authors present a case of five-beat clonus in the absence of a central nervous system pathologic condition. As data obtained from the literature demonstrate, clinicians should be aware that multi-beat clonus is not always associated with upper motor neuron lesions. PMID- 2567348 TI - Effects of ICI 169,369, a selective serotonin2 antagonist, in electrophysiological tests predictive of antipsychotic activity. AB - Extracellular single unit recording techniques were used to compare the effects of ICI 169,369, a selective serotonin2 receptor antagonist, with the reference antipsychotic (AP) agents clozapine and haloperidol, in electrophysiological tests that may predict AP activity. ICI 169,369 was found to reverse the inhibitory actions of amphetamine on A9 and A10 dopamine (DA) neurons, a common property shared by other AP drugs, and was comparable in potency to clozapine. In cell population studies, acute treatment with ICI 169,369 (at a low dose only) and clozapine selectively increased the number of spontaneously active A10 DA cells, which was found to correlate with the ability of both these drugs to cause depolarization inactivation (DI) of A10 DA cells after chronic administration. Interestingly, chronic treatment with ICI 169,369 also caused a significant increase in the number of actively discharging A9 DA cells, an effect not predicted on the basis of the acute data. A similar effect was noted for clozapine, although the magnitude did not reach statistical significance. This profile of activity was unlike that of haloperidol, which acutely caused a nonselective increase in the number of active A9 and A10 DA cells, associated with the ability of this agent to cause DI of both A9 and A10 DA cells after chronic treatment. Inasmuch as DI of A10 DA cells may be correlated with AP efficacy whereas DI of A9 DA cells may predict the ability of an AP to cause extrapyramidal side effects, ICI 169,369, like clozapine, may be a potential AP with a reduced likelihood for producing extrapyramidal side effects. PMID- 2567349 TI - Synergistic alpha-1 and alpha-2 adrenergic stimulation of rat proximal nephron Na+/H+ exchange. AB - Both alpha-1 and alpha-2 adrenoceptors have been localized to the renal cortex, with the majority of binding sites on the proximal tubule. Because the major regulator of Na+ uptake into the proximal tubule is the Na+/H+ exchanger, and because alpha-1 and alpha-2 adrenoceptors stimulate it in other tissues, we tested the hypothesis that both alpha adrenoceptor subtypes can increase Na+ uptake into the proximal nephron by stimulating the Na+/H+ antiporter. Enhancement of Na+ transport by agonists was studied in isolated rat proximal tubules by determining the uptake of 22Na that was suppressible by the Na+/H+ inhibitor, 5-(N-ethyl-N-isopropyl)amiloride (EIPA). The phorbol ester, phorbol-12 myristate-13-acetate, (0.1 microM), directly stimulated the antiporter through protein kinase C and increased EIPA-suppressible 22Na uptake 250% above control. The alpha-1 adrenoceptor agonists, cirazoline and phenylephrine, in addition to the mixed agonist, norepinephrine, maximally stimulated uptake by 226 to 232% at 1 microM concentrations. alpha-2 agonists produced a range of maximal stimulations at 1 microM from 65% with guanabenz to 251% with B-HT 933. Increases in 22Na uptake by agonists were inhibited by selective adrenergic antagonists and by EIPA. The drugs did not change the EIPA-resistant component of 22Na uptake. Inasmuch as the adrenoceptor subtypes likely stimulated Na+/H+ exchange by differing intracellular pathways impinging upon common transport steps, we examined whether simultaneous stimulation of both pathways was additive. Submaximal concentrations (5 nM each) of alpha-1 and alpha-2 adrenoceptor agonists in combination synergistically enhanced 22Na uptake to a level similar to 1 microM concentrations of adrenoceptor agonists alone or in combination.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567350 TI - Multiplicative interaction between intrathecally and intracerebroventricularly administered mu opioid agonists but limited interactions between delta and kappa agonists for antinociception in mice. AB - Simultaneous action of morphine on supraspinal and spinal sites produces a multiplicative interaction for antinociception which may be important for the analgesia produced by systemically administered morphine. The purpose of this study was to see whether other agonists with more receptor selective opioid actions than morphine would also produce this multiplicative interaction. DAMPGO (Tyr-D-Ala2-Gly-NMePhe4-Gly-ol5), DPDPE (D-Pen2, D-Pen5, enkephalin) and U50 488H, opioid agonists highly selective for mu, delta and kappa receptors, respectively, were administered alone i.c.v. or intrathecally (i.t.) or in combination (i.c.v. plus i.t.) to determine ED50 values for the tail-flick response in mice. These ED50 values were examined isobolographically in relation to the theoretical additive ED50 values by the potency ratio method. First, DAMPGO given i.cv and i.t. was similar to morphine, indicating that simultaneous supraspinal and spinal mu agonist administration produce the multiplicative interaction. Second, concurrent administration of DPDPE or U50,488H, i.c.v. and i.t., as well as cross-over combinations of DPDPE at one and U50,488H at the other site, produced additive interactions only. The multiplicative interaction was a property characteristic of mu but not delta and kappa agonists. Based on the similarity between morphine and DAMPGO, it was postulated that both mu agonists act on redundant descending pain inhibitory pathways to produce multiplication. A second mechanism for multiplicative interaction was based on the difference between DAMPGO and morphine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567351 TI - Acute extrapyramidal syndrome in Cebus monkeys: development mediated by dopamine D2 but not D1 receptors. AB - The present study assessed the role of dopamine D1 and D2 receptors in the production of an extrapyramidal syndrome (EPS) in Cebus apella monkeys. Previous studies have shown the development of EPS in both old and new world monkeys with haloperidol administration. We now report that repeated weekly administration of a selective D1 antagonist, SCH 23390, does not produce this syndrome in cebus monkeys. Cebus monkeys were treated with either vehicle (n = 6), the specific D2 antagonist haloperidol (0.3 mg/kg p.o., n = 9) or the specific D1 antagonist SCH 23390 (10.0 mg/kg p.o., n = 9) once a week for approximately 1 year and behavioral effects were observed and scored. The drug doses used in this study produced similar sedative scores when given acutely and sedation increased over the first 12 weeks of the study for both treatment groups. However, by the 12th week of dosing with haloperidol all the monkeys showed a profound EPS characterized by limb extensions, head pushing, tongue protrusions and sometimes severe biting movements. In contrast, none of the SCH 23390-treated monkeys showed any abnormal movements, suggesting D1 antagonists have a low EPS side effect liability. The profile of the incidence of EPS seen with classical neuroleptic drugs in cebus monkeys and their blockade of EPS by anticholinergic drugs mimics the profile seen in humans. The models presented appear to be predictive of the production of the EPS in humans and could be used to screen neuroleptics for EPS liability. Furthermore, the EPS is probably due to the selective blockade of dopamine D2 receptors with its associated enhancement of cholinergic neurotransmission.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567352 TI - Electrophysiological characteristics and pharmacological responsiveness of midbrain dopaminergic neurons of the aged rat. AB - The extracellular electrophysiological activities of A9 and A10 dopaminergic (DA) neurons were examined in chloral hydrate-anesthetized young (3 months) and aged (24-28 months) Fischer 344 male rats. Average firing rate, firing rate distribution and the incidence of spontaneous activity (cells per electrode track) did not change in either region as a function of age. Next, the inhibitory effects of several DA agonists on the firing rates of antidromically identified nigrostriatal DA cells were compared in young and old rats. The inhibitory potencies of i.v. apomorphine (D1/D2 agonist), quinpirole (D2 agonist) and d amphetamine (indirect agonist) did not change with age. Similarly, the inhibitory effects of microiontophoretically applied dopamine were unaltered in aged rats. Stimulation of the sciatic nerve resulted in brief periods of inhibition of the activity of nigrostriatal DA neurons in both the young and the old age groups. The duration of this inhibitory response was, however, enhanced markedly in the old rats. The D1 agonist SKF 38393 enhanced the duration of the inhibitory effect of sciatic nerve stimulation in both young and old rats. These results suggest that, although no gross differences in the activity of DA cells in young and old rats were evident, subtle changes in the regulation of the activity of these neurons may occur with age. PMID- 2567353 TI - Species differences in the localization and number of CNS beta adrenergic receptors: rat versus guinea pig. AB - The localization and number of beta adrenergic receptors were directly compared in the brains of rats and guinea pigs. The time course of association and saturability of [125I]cyanopindolol (CYP) binding to slide-mounted tissue sections was similar in rats (Kd = 17 pM) and guinea pigs (Kd = 20 pM). The beta 1 and beta-2 receptor subtypes were examined through the use of highly selective unlabeled receptor antagonists, ICI 118,551 (50 nM) and ICI 89,406 (70 nM). Dramatic species differences between rats and guinea pigs were observed in the neuroanatomical regional localization of the beta adrenergic receptor subtypes. For example, in the thalamus prominent beta-1 and beta-2 receptor populations were identified in the rat; however, the entire thalamus of the guinea pig had few, if any, beta adrenergic receptors of either subtype. Hippocampal area CA1 had high levels of beta-2 adrenergic receptors in both rats and guinea pigs but was accompanied by a widespread distribution of beta-2 adrenergic receptors only in rats. Quantitative autoradiographic analyses of 25 selected neuroanatomical regions 1) confirmed the qualitative differences in CNS beta adrenergic receptor localization, 2) determined that guinea pigs had significantly lower levels of beta adrenergic receptors than rats and 3) indicated a differential pattern of receptor subtypes between the two species. Knowledge of species differences in receptor patterns may be useful in designing effective experiments as well as in exploring the relationships between receptor and innervation patterns. Collectively, these data suggest caution be used in extrapolation of the relationships of neurotransmitters and receptors from studies of a single species. PMID- 2567355 TI - Multidrug resistance: molecular biology and clinical relevance. PMID- 2567356 TI - Multidrug resistance: P-glycoprotein and its allies--the elusive foes. PMID- 2567354 TI - Fertility in a male with trisomy 21. AB - We review the published reports on reproduction in cases of non-mosaic trisomy 21 (Down's syndrome) and present the first fully documented case of a non-mosaic male with Down's syndrome fathering a pregnancy, a fact which has important implications in the light of caring for these people in the community. PMID- 2567357 TI - Laparoscopy: its selected use in patients with unilateral nonpalpable testis after human chorionic gonadotropin stimulation. AB - Laparoscopy has been used to help evaluate patients with a unilateral nonpalpable testis. This procedure can be performed quickly just before exploration with the patient under the same anesthetic. With laparoscopy it has been possible to localize either a testis or the course of the spermatic vessels in 100 per cent of the patients. Preoperative knowledge of testis location is helpful to plan the location of the incision as well as the type of repair. The incidence of vanishing testis in our series is much higher than that reported previously. This difference is attributed to a careful examination with the patient under anesthesia and preoperative treatment with human chorionic gonadotropin, which made many testes palpable. These results indicate that laparoscopy can be performed safely and quickly, and that it is helpful to manage patients with a unilateral nonpalpable testis. PMID- 2567358 TI - Symptomatic urogenital manifestations of polyarteritis nodosa and Churg-Strauss angiitis: analysis of 8 of 165 patients. AB - To establish the clinical features and evolution of ureteral and testicular manifestations of polyarteritis nodosa, 8 cases among a group of 165 patients were reviewed. Urogenital manifestations are rare and were related to hepatitis B virus markers in 5 of the 8 patients. Malignant hypertension was present in 6 of 8 patients and renal insufficiency related to renal infarctions was noted in 5. Prognosis is good and regression generally is obtained with steroid therapy or with plasma exchange and vidarabine when polyarteritis is related to hepatitis B virus. In some cases ureteral stenosis may require an operation in association with steroids and/or immunosuppressive treatments. PMID- 2567359 TI - Scientists ponder when, why of emerging viruses. PMID- 2567360 TI - [Remarks on some analgesics used in the pain clinic]. AB - The mechanism of action of aspirin as an analgesic is an inhibition of biosynthesis of prostaglandins. Thus the site of action has been believed to be peripheral. However, when aspirin is injected intra- thecally, it produces an analgesic effect. Aspirin has a membrane-stabilizing effect and it is used locally for the treatment of post- herpetic neuralgia. Epidural opioids are frequently used for the management of post-operative pain or cancer pain. Pharmacokinetic studies have shown that delayed respiratory depression results from migration of morphine in the cerebrospinal fluid to the brain. Peak concentrations of morphine near the brain stem occur about 3 hours after lumbar epidural injection, whereas lipophilic opioids such as meperidine, peak concentration occur within 30 to 60 minutes. The clearance from cerebrospinal fluid of lipophilic opioids is more rapid than that of morphine. Besides opioids, alpha 2 receptor agonists such as clonidine also have analgesic action when administered into the epidural space. Somatostatin is one of many neuropeptides found in the spinal cord. It has dual action: a mediation of thermal nociception and a general antinociceptive action. When somatostatin is administered intrathecally or epidurally, it produces analgesic effect and its efficacy appears to be equal to that of morphine. PMID- 2567361 TI - [Sialylated Lewis(x) and sialylated Lewis(a) as tumor-associated carbohydrate antigens in sera of patients with gastric cancer--usefulness of combination assay of these two antigens]. AB - Sialylated Lewis(x) and sialylated Lewis(a) antigens, which are structural isomer each other, were serologically tested to determine the clinicopathological differences and correlation between two antigens, and were also compared with those of serum CEA in 141 patients with gastric cancer. Percent positives in sialylated Lewis(a), CEA and sialylated Lewis(x) showed 26.9%, 21.8% and 17.0% respectively. Percent positive of sialylated Lewis(x) correlated with degree of liver metastasis and peritoneal dissemination. Therefore, it is emphasized that the role of sialylated Lewis(x) is characteristics of detecting a presence of liver metastasis and/or peritoneal dissemination. On the other hand, sialylated Lewis(a) showed high percent positive in cases which had lymph nodes metastasis and mucinous or papillary adenocarcinomas. These two carbohydrate antigens indicated no correlation (r = 0.062) in the spectrum of sera. It is suggested that combination assay of sialylated Lewis(x) and sialylated Lewis(a) is useful in detecting many gastric cancer patients including CEA-negative one. PMID- 2567363 TI - [Impression of the Second Asia and Pacific Nurses' Convention held at Singapore]. PMID- 2567362 TI - [Salicylazosulfapyridine-induced male infertility in a patient with ulcerative colitis]. PMID- 2567364 TI - [A study on acute renal damage from percutaneous nephrolithotomy assessed by urinary NAG and gamma-GTP activities]. AB - We performed percutaneous nephrolithotomy (PNL) on 49 patients between May, 1986 and March, 1987. To investigate acute renal damage from PNL, we measured urinary NAG (N-acetyl-beta-D-glucosaminidase) and gamma-GTP (gamma-glutamyl transpeptidase) activities before PNL and for 6 days after PNL in 24 patients. The NAG activities elevated beyond normal level in 23 patients and gamma-GTP activities in 15 patients. NAG activities showed a peak level in the third day after PNL and gamma-GTP activities in the next day of PNL. After the peak both enzyme activities got down gradually. There was no difference in NAG and gamma GTP activities between nephrostomy and lithotripsy in 2 staged patients. And the intrapelvic pressure during operation was at the normal level in 5 patients. Therefore, we think that the cause of NAG and gamma-GTP activity elevation is a mechanical damage, not an influence of the irrigation fluid. Large stones, long operation time and 2 stage procedure were the factors that produced high enzyme activities, because, we guess, the frequency of mechanical damages to the kidney increase in such cases. Postoperative pyrexia caused a slight increase in enzyme activities but preoperative hydronephrosis exerted no influence on both enzyme activities. We also measured creatinine clearance before and after PNL but no significant change was obtained. PNL causes only slight mechanical damage to the operated kidney which is reversible when assessed by NAG and gamma-GTP activities and the glomerular function is not affected. Therefore, we conclude that PNL is a safe treatment for upper urinary tract stones. PMID- 2567365 TI - [Experimental studies on urinary incontinence. I. Effects of beta-adrenoceptor agonists on the contractile activities and intracellular cAMP levels of rabbit, canine and human urinary bladder smooth muscle]. AB - Beta-adrenoceptor subtypes in detrusor and trigonal muscle of rabbit, canine and human urinary bladder were studied by in vitro techniques recording contractile force and by measuring intracellular cAMP content. The results of contractile studies were correlated with the intracellular cAMP levels. In the rabbit bladder, only beta 2-receptors were found in the detrusor though both beta 1- and beta 2-receptors were found in the trigonal muscle. In contrast, both beta 1 and beta 2-receptors were present in both detrusor and trigonal muscle of canine bladder. Human detrusor had only beta 2-receptors. Thus there are species differences in beta-adrenoceptor subtypes in urinary bladder smooth muscle. PMID- 2567366 TI - [Experimental studies on urinary incontinence. II. Function and distribution of alpha 1- and alpha 2-adrenoceptors in male and female rabbit urethral smooth muscle]. AB - The effects of various alpha-adrenergic agonists and antagonists on the contractile properties of the isolated urethra of male and female rabbits were examined and the population of alpha 1- and alpha 2-adrenoceptors, using radioligand receptor binding techniques, was also quantified to define the functional role of postsynaptic alpha-adrenoceptors in urethral smooth muscle. Norepinephrine, epinephrine (non-specific alpha-adrenergic agonists) and phenylephrine (alpha 1-adrenergic agonists) induced increases in contractile force in both the male and female urethra. Clonidine (alpha 2-adrenergic agonist) caused a relatively large contractile response in the female urethra but only a small contractile response in the male urethra. Receptor binding studies indicated that the male urethra contains almost equal amounts of alpha 1- and alpha 2-adrenoceptors (32 vs. 34 fmol/mg protein, respectively) whereas the female urethra contains a significantly greater density of alpha 2- than alpha 1 adrenoceptors (122 vs. 34 fmol/mg protein, respectively). Our studies have indicated that: (1) both alpha 1-and alpha 2-adrenoceptors cause contractile responses in male and female rabbit urethra; (2) the greater response to alpha 2 agonist in female than male urethra is correlated with a higher density of alpha 2-adrenoceptors in this tissue. PMID- 2567367 TI - [Combined treatment with selective beta1 adrenoblockaders and beta2 adrenostimulants in patients with myocardial infarction associated with chronic obstructive lung diseases]. AB - The effect of selective beta 1-adrenoblocker metoprolol, used alone or in combination with selective beta 2-adrenostimulant terbutaline, on hemodynamic and gas exchange parameters was examined in 30 patients with acute myocardial infarction (MI) and attendant chronic pulmonary obstructive diseases (CPOD). Severe gas exchange and intracardiac hemodynamic disorders, conducive to right ventricular decompensation, were demonstrated in this category of patients. Selective beta 1-adrenoblockers can be used as part of combined treatment for MI under the monitoring of external respiration parameters in patients with mild bronchial obstruction and moderate hypoxemia, whereas in MI patients with severe respiratory insufficiency and gas exchange disorders, the treatment with beta 1 adrenoblockers can aggravate those. Combined use of metoprolol and brikanil produces a number of favorable ventilatory and hemodynamic effects, conducive to the stabilization of clinical condition in MI patients with attendant CPOD and limits the risk of complications. PMID- 2567368 TI - Characterization of the antihistamines tripelennamine, methapyrilene, and thenyldiamine and their N-oxides by thermospray mass spectrometry. AB - This study describes an investigation of the thermospray (TS) mass spectrometric analysis of tripelennamine, methapyrilene, thenyldiamine, and their N-oxide derivatives. These compounds were analyzed by direct injection TS mass spectrometry in the column bypass mode and with 0.1M ammonium acetate/methanol (80:20, v/v) as the mobile phase. Typically, the parent antihistamines produced only [MH]s ions under these conditions. The N-oxides provided strong [MH]s ions and multiple fragment ions. A scheme to explain the fragmentation patterns is proposed. PMID- 2567369 TI - [The 5th World Congress on AIDS: multidisciplinary discussions on waiting for a break-through in laboratory research]. PMID- 2567370 TI - Rapid, sensitive diagnosis of malaria based on ribosomal RNA. AB - A system for the sensitive and accurate diagnosis of all four species of malaria parasite that are pathogenic in man has been developed. It involves hybridisation of oligonucleotides complementary to species-specific regions of the RNA of the parasite small ribosomal subunit followed by autoradiography. The method retains its specificity even under conditions of very low stringency similar to those that will occur in field diagnosis. Direct application of treated blood to nylon is possible and consistently results in the detection of fewer than 10 parasites by autoradiography in an overnight exposure. The target rRNA is stable even in dehydrated cells. PMID- 2567371 TI - Chronic cough: eosinophilic bronchitis without asthma. AB - Sputum cell-counts were studied in 7 non-smokers with corticosteroid-responsive chronic cough productive of sputum and 8 smokers with a clinical diagnosis of chronic bronchitis, all of whom had normal lung function tests and methacholine airway responsiveness, and in 10 non-smokers with asthma, examined during an exacerbation. Sputum from asthmatic patients and subjects with corticosteroid responsive cough contained eosinophils and metachromatic cells. Macrophages were by far the dominant cell type in sputum from subjects with chronic bronchitis. Airway inflammation with eosinophils and metachromatic cells is not always accompanied by increased airway responsiveness, and current definitions of obstructive airways disease may need to be revised. PMID- 2567372 TI - Prevention of Pneumocystis carinii pneumonia relapse by pentamidine aerosol in zidovudine-treated AIDS patients. AB - To examine the efficacy and tolerance of pentamidine aerosol in the prevention of Pneumocystis carinii pneumonia (PCP) relapse in patients with the acquired immunodeficiency syndrome (AIDS) being treated with zidovudine, 51 patients who had had an episode of PCP in the previous 5 months were enrolled in a randomised controlled study. 25 patients (group I) received pentamidine mesylate aerosol (4 mg/kg every 2 weeks for the first month then monthly) and zidovudine, and 26 patients (group II) zidovudine alone. 3 group I patients withdrew from pentamidine therapy prematurely and were excluded from the analysis of efficacy. Relapses of PCP occurred in 2 out of 22 (9%) group I patients and in 16 out of 26 (61%) group II patients after a mean follow-up of 10 and 8.7 months, respectively. The two groups differed significantly (p less than 0.0001) in proportions without relapse. They did not differ in proportions surviving. Bronchial intolerance was common (47%); no systemic side-effects of pentamidine were observed. Pentamidine aerosol thus seems to be effective in preventing PCP relapses in AIDS patients on zidovudine. The early termination of the trial prevented assessment of the long-term efficacy and safety of pentamidine given by aerosol. PMID- 2567373 TI - Protein-losing enteropathy associated with Clostridium difficile infection. AB - A commercially available radial immunodiffusion assay was used to measure serum alpha-1-antitrypsin levels in stool samples from subjects aged over 60 years as a marker of protein-losing enteropathy. alpha 1-antitrypsin was found in all of 12 patients with colonoscopy-confirmed pseudomembranous colitis, 6 of 14 (43%) patients with Clostridium difficile diarrhoea without pseudomembranes, 6 of 12 (50%) nursing-home patients culture-positive for Cl difficile but negative for its cytotoxin, and none of 15 healthy control subjects. It is concluded that serum protein loss into the gastrointestinal tract can occur as a result of Cl difficile infection, that its presence correlates with the severity of disease, and that it may occur even in the absence of diarrhoea. The diagnosis of protein losing enteropathy should be considered for all patients with Cl difficile infection, particularly elderly nursing-home patients, in whom the risk of Cl difficile disease and the frequency of severe malnutrition are high. PMID- 2567374 TI - Sequential changes in serum insulin concentration during development of non insulin-dependent diabetes. AB - Changes in serum insulin concentrations during deterioration of glucose tolerance were studied in 81 Pima Indians who worsened from normal to impaired glucose tolerance (IGT); 44 who changed from IGT to non-insulin-dependent diabetes mellitus (NIDDM); 27 who were seen at diagnosis of NIDDM and 1.4-8.5 years later; and 11 subjects who were seen at each of these stages. When their glucose tolerance was normal, subjects who later developed NIDDM had higher fasting and post-load insulin concentrations than controls of similar age and body mass index who did not become diabetic. Onset of IGT or NIDDM was associated with a further increase in fasting insulin concentrations, although a deterioration from IGT to NIDDM was associated with little change in insulin responses to oral glucose in spite of increased blood glucose. After the onset of NIDDM, both fasting and post load insulin concentrations diminished. These longitudinal data show that, as glucose tolerance worsens, insulin and glucose concentrations in individuals follow the inverted-U-shaped relation previously reported in cross-sectional population studies. PMID- 2567375 TI - TU or not TU. PMID- 2567376 TI - Sunlight, DNA repair, and skin cancer. PMID- 2567377 TI - Routine H2-receptor antagonists before elective surgery? PMID- 2567378 TI - Low-dose vitamin D analogues for renal osteodystrophy. PMID- 2567379 TI - Stress and panic. PMID- 2567380 TI - A youthful centenarian. PMID- 2567381 TI - Discrepancies between the effects of coronary reperfusion on survival and left ventricular function. PMID- 2567382 TI - Ultrasound-guided core biopsy. AB - Between February, 1986, and September, 1988, 404 biopsies have been done, under ultrasound guidance, with a cutting needle fired by a spring-loaded device. Sensitivity for a diagnosis of malignancy was 92.7%; overall accuracy was 94.1%; the predictive value of a positive result was 100% and the complication rate was low. An exact diagnosis could be made from histological examination of the specimen obtained from 98% of positive tumour biopsies. This technique is a simple and safe way to obtain specimens for accurate histological diagnosis. PMID- 2567383 TI - Stroke distance in acute myocardial infarction: a simple measurement of left ventricular function. AB - Stroke distance was measured by means of doppler ultrasound in 100 consecutive patients with acute myocardial infarction. Mean stroke distance on days 1-5 after infarction was 72% of the predicted normal value for age and rose to 77% of normal by days 6-10. Stroke distance was lower after anterior (65%) than inferior infarction (75%), and was negatively correlated with peak serum aspartate aminotransferase (r = 0.47, p less than 0.001). Of 16 patients whose stroke distance on days 1-5 was below 50% of normal for age, 4 died, all within the first week. Of 84 patients whose stroke distance was 50% of normal or higher, only 1 died (at 13 days). The simple bedside measurement of stroke distance offers a practical method of monitoring left ventricular function in acute myocardial infarction: the measurement may have prognostic value. PMID- 2567384 TI - Peptide regulatory factors in the nervous system. PMID- 2567386 TI - Hormone dependency of meningiomas. PMID- 2567385 TI - Medicolegal investigation of political killings in El Salvador. PMID- 2567387 TI - In-utero transplantation of stem cells in bare lymphocyte syndrome. PMID- 2567389 TI - New treatment for endometriosis. PMID- 2567388 TI - Indications for flecainide. PMID- 2567390 TI - Symmetrical peripheral gangrene. PMID- 2567391 TI - Alpha-interferon treatment of patient with hyper IgE syndrome. PMID- 2567392 TI - Fine-needle aspiration of tumours. PMID- 2567393 TI - Cervical cancer in young Americans. PMID- 2567394 TI - Marital status and cervical cancer in young women. PMID- 2567395 TI - Ovulation induction and neural tube defects. PMID- 2567396 TI - Intermittent flumazenil and benzodiazepine tolerance: discouraging findings in rats. PMID- 2567397 TI - UVA filters in sunscreen preparations. PMID- 2567398 TI - Alcohol swabs and venepuncture. PMID- 2567399 TI - Nebulised budesonide in severe childhood asthma. PMID- 2567400 TI - Subcutaneous erythropoietin and peritoneal dialysis. PMID- 2567401 TI - Intraperitoneal erythropoietin. PMID- 2567402 TI - Anti-CD3 antibody for autoimmune disease, a cautionary note. PMID- 2567403 TI - Measles-specific nucleotide sequences and autoimmune chronic active hepatitis. PMID- 2567404 TI - Detection of measles virus RNA in paraffin-embedded tissue. PMID- 2567405 TI - Fortification of foods for refugees. PMID- 2567406 TI - Rationing of medical care. PMID- 2567407 TI - Advertising drugs to the public. PMID- 2567408 TI - Clicking hip: a risk factor for congenital hip dislocation. PMID- 2567410 TI - Feminism in publishing. PMID- 2567409 TI - Cost-benefit of self-prescribing. PMID- 2567411 TI - Screening for asymptomatic bacteriuria in the elderly. PMID- 2567412 TI - Hypoglycaemic unawareness and human insulin. PMID- 2567413 TI - Clonidine for short stature. PMID- 2567414 TI - Gastrointestinal atresias and borreliosis. PMID- 2567415 TI - Clinical problems during fast of Ramadan. PMID- 2567416 TI - Familial osteoarthrosis and type II collagen gene. PMID- 2567417 TI - Sensitive thyrotropin measurements: utility and futility. PMID- 2567418 TI - Aluminium-to-creatinine ratios in infancy. PMID- 2567419 TI - Frailty selection and HIV. PMID- 2567420 TI - Diclazuril for Isospora belli infection in AIDS. PMID- 2567422 TI - Reduced platelet aggregation in long-distance runners. PMID- 2567421 TI - Debrisoquine hydroxylation phenotypes in healthy volunteers. PMID- 2567423 TI - Menkes' disease: a disorder of zinc metabolism? PMID- 2567424 TI - Iron, asbestos, and carcinogenicity. PMID- 2567425 TI - Nausea and vomiting after general anaesthesia. PMID- 2567426 TI - Fatal reaction to suxamethonium: case for screening by radioallergosorbent test? PMID- 2567427 TI - Ecological impact of ampicillin and cefuroxime in neonatal units. AB - The relation between local antibiotic consumption and faecal carriage of beta lactam-resistant strains of Escherichia coli and Klebsiella spp was studied in 953 children discharged from twenty-two neonatal units. An increased rate of carriers of Klebsiella spp strains resistant to ampicillin, cefuroxime, and cephalexin and of E coli strains resistant to ampicillin was associated with the use of ampicillin, with or without gentamicin. Ampicillin and cephalosporin (86% cefuroxime) consumptions were inversely related, which probably explained the paradox that cephalosporin use (and therefore less ampicillin use) was negatively correlated with the carriage of Klebsiella spp strains resistant to ampicillin, cefuroxime, and cephalexin. The ecological impact of antibiotics was much the same among treated and untreated babies in each unit. The findings show a greater risk of drug resistance due to a related agent rather than to the drug itself and a clear indirect impact of an antibotic on the microflora of untreated patients. Ampicillin-based regimens are more likely than cefuroxime to produce drug resistant strains in the newborn. PMID- 2567428 TI - DNA restriction fragment length polymorphisms as markers of familial coronary heart disease. AB - 246 of 713 men aged 30 to 59 years had first-degree relatives who had had coronary heart disease (CHD) by the age of 60; men with a family history of CHD were twice as likely to have CHD themselves, compared with those without such a family history. At least 75% of the difference was accounted for by CHD in men with minor alleles of 4 restriction fragment length polymorphisms (RFLPs) in the region of the apolipoprotein (apo) AI and apo CIII genes. The RFLPs were identified with the restriction enzymes XmnI, PstI, MspI, and SacI. Each polymorphism has two alleles (major and minor), designated X1 and X2, P1 and P2, M1 and M2, and S1 and S2, respectively. In men with any one of the minor alleles, a family history of CHD was associated with a 234% increase in CHD prevalence. In men with major alleles only, a family history of CHD was not associated with any significant increase in CHD. The effect of the minor alleles was not significantly altered when plasma lipids and other variables except for age were taken into account. The association between the minor RFLP alleles and polymorphic gene variants (probably the apo AI, apo CIII, or both genes) which enhance liability to CHD accounted for almost 20% of total CHD in this population. PMID- 2567429 TI - Serological diagnosis of salmonella infections by enzyme immunoassay. AB - An enzyme immunoassay (EIA) for the detection and measurement of serum IgM, IgG, and IgA antibodies to salmonella was developed with commercially available lipopolysaccharides (LPSs) of Salmonella typhimurium and S enteritidis combined as antigen. Of 130 sera from patients with culture-confirmed salmonella infections, 115 (88.5%) were positive in this assay. The classic Widal agglutination test was positive in only 50 (38.5%) cases. This EIA method offers a substantial advance in the serological diagnosis of acute salmonella infections; it detects antibodies to the salmonellae of groups B and D, which constitute about 70% of culture-positive cases of human salmonellosis. Antibodies to other salmonellae are also detected. This EIA is particularly valuable for the detection of salmonella antibodies during post-infectious complications when isolation of the organism is often no longer possible. PMID- 2567430 TI - Variable expression of P fimbriae in Escherichia coli urinary tract infection. AB - Fresh urinary isolates were examined by immunofluorescence with polyclonal rabbit antibodies against type 1 and P fimbriae. This procedure showed P-fimbriate Escherichia coli in 22 of 24 samples from patients with asymptomatic bacteriuria, 24 of 26 samples from patients with cystitis, and 6 of 6 samples from patients with pyelonephritis. Type 1 fimbriae were expressed by less than 40% of isolates in all three groups. There was no relation between the presence of symptoms or the site of infection and fimbrial expression, of P or type 1, by bacteria adherent to freshly isolated uroepithelial cells. PMID- 2567432 TI - Low cholesterol and increased risk. PMID- 2567431 TI - Nebulised salbutamol with and without ipratropium bromide in acute airflow obstruction. AB - 103 patients with acute airflow obstruction (56 asthma, 47 chronic obstructive pulmonary disease [COPD]) completed a double-blind trial of nebulised bronchodilator treatment in a hospital accident and emergency department. Each patient was randomised to receive either 10 mg of salbutamol nebuliser solution in 2 ml of saline or 10 mg of salbutamol in 2 ml (0.5 mg) of preservative-free ipratropium bromide. Peak flow rate (PFR) was recorded before treatment and 1 hour after beginning nebulised treatment. In 23 asthmatic patients given salbutamol alone PFR rose by a mean 31% 1 hour after treatment whereas in 33 such patients given combined treatment it rose by a mean 77% (95% confidence interval for the difference 8-84%). Patients whose PFR was below 140 l/min at entry gained maximum benefit from the combined treatment. For COPD patients the PFR rise was almost identical for both treatments. In acute asthma the immediate PFR response to a mixture of salbutamol and ipratropium bromide was better than the response to nebulised salbutamol alone. For COPD patients, the two treatments were of equal benefit. PMID- 2567433 TI - Aplasia leukaemia syndrome. PMID- 2567434 TI - Medical education goes to market. PMID- 2567435 TI - Achilles tendon rupture. PMID- 2567436 TI - Fax of life. PMID- 2567437 TI - Effects on weight and metabolic rate of obese women of a 3.4 MJ (800 kcal) diet. AB - 103 obese women (mean [SD] Quetelet's index [weight/height2] 38 [8] kg/m2) were admitted to a metabolic ward and were kept strictly to a diet providing 3.4 MJ (800 kcal) daily for 3 weeks. Body weight was measured daily and fasting resting metabolic rate (RMR) on days 1, 7, and 21. Both weight and RMR fell more rapidly in the first week than later. The thermic effect of feeding fell immediately on the lower energy intake, and there was an adaptive reduction of about 6% in RMR in week 1. After 3 weeks, the average weight loss was 4.9 (1.2) kg (about 5% of initial weight) and the average fall in RMR 8.8%. If after substantial weight loss a woman eats just enough to maintain energy balance the adaptive reduction in metabolic rate is restored to normal, and the thermic effect of feeding is restored in proportion to the new energy intake, but total energy requirements remain less than in the obese state to the extent that fat-free mass has been reduced. Thus, an obese woman who reduces weight by 30% over a year will thereafter have requirements for weight maintenance which are reduced by about 15%. PMID- 2567438 TI - Peptide regulatory factors in non-malignant disease. PMID- 2567439 TI - Controlled trial of acupuncture for severe recidivist alcoholism. AB - In a placebo-controlled study, 80 severe recidivist alcoholics received acupuncture either at points specific for the treatment of substance abuse (treatment group) or at nonspecific points (control group). 21 of 40 patients in the treatment group completed the programme compared with 1 of 40 controls. Significant treatment effects persisted at the end of the six-month follow-up: by comparison with treatment patients more control patients expressed a moderate to strong need for alcohol, and had more than twice the number of both drinking episodes and admissions to a detoxification centre. PMID- 2567440 TI - Side-effects of ivermectin in treatment of onchocerciasis. AB - In a prospective study to determine the tolerance for and safety of ivermectin therapy for onchocerciasis in a hyperendemic area in Sierra Leone, 28 (32%) of 87 patients had adverse reactions that required treatment with acetylsalicylic acid and antihistamines, but none of the observed adverse reactions were considered life-threatening. A significant relation was found between the frequency and severity of side-effects and the degree of parasite infestation, as quantified by the skin-snip counts. Free administration of ivermectin to severely infected onchocerciasis patients is not recommended without some form of medical supervision. PMID- 2567441 TI - The World Medical Association and South Africa. PMID- 2567442 TI - Long cross-clamp time with warm heart surgery. PMID- 2567443 TI - Childhood leukaemia mortality before 1970 among populations near two US nuclear installations. PMID- 2567444 TI - Apgar score and audit. PMID- 2567445 TI - Gender and survival. PMID- 2567446 TI - Hazards of routine endotracheal suction in the neonatal unit. PMID- 2567447 TI - Booking scan dates discrepancy and repeat scanning. PMID- 2567449 TI - Myopathy as possible side-effect of cyclosporin. PMID- 2567448 TI - Treatment of Parkinson's disease with novel dopamine D2 agonist SK&F 101468. PMID- 2567450 TI - Hyperkalaemia--silent and deadly. PMID- 2567451 TI - Serotonin receptors, buspirone, and premenstrual syndrome. PMID- 2567452 TI - Alpha-blockade for hypertension. PMID- 2567453 TI - Verocytotoxin-producing Escherichia coli O2:H5 isolated from patients with ulcerative colitis. PMID- 2567454 TI - Breast cancer and oral contraceptives. PMID- 2567455 TI - Proportion of newly obese and chronic obese at different ages. PMID- 2567456 TI - In-vitro study of immune responses in loin pain/haematuria syndrome. PMID- 2567457 TI - Nursery-acquired asymptomatic B hepatitis. PMID- 2567458 TI - Handwashing and horizontal spread of viruses. PMID- 2567459 TI - Induction of hyperthyroidism by interferon-alpha-2b. PMID- 2567460 TI - Pyroglutamicaciduria from vigabatrin. PMID- 2567461 TI - Alzheimer's disease in antacid users. PMID- 2567463 TI - Giving permission. PMID- 2567462 TI - Is susceptibility to Campylobacter pylori infection genetically determined? PMID- 2567464 TI - Good clinical practice in clinical research. PMID- 2567465 TI - Surgical examinations. PMID- 2567466 TI - Continuous infusion of ceftazidime in cystic fibrosis. PMID- 2567467 TI - Can Wilson's disease patients be decoppered? PMID- 2567468 TI - HLA-DP based resistance to Hodgkin's disease. PMID- 2567469 TI - Carbimazole-induced cramps. PMID- 2567470 TI - Carcinoid tumours and endocrine cell hyperplasia. PMID- 2567471 TI - Parvovirus infection, leukaemia, and immunodeficiency. PMID- 2567472 TI - High frequency of Epstein-Barr virus genome in HIV-positive patients with Hodgkin's disease. PMID- 2567473 TI - Inhaled steroids and recurrent wheeze after bronchiolitis. PMID- 2567474 TI - Failure of pyrimethamine-clindamycin combination for prophylaxis of Pneumocystis carinii pneumonia. PMID- 2567475 TI - Prevention of AIDS. PMID- 2567476 TI - Adjuvant chemotherapy for stomach cancer. PMID- 2567477 TI - Cardiac standstill induced by prostaglandin pessaries. PMID- 2567478 TI - Vth international conference on AIDS. PMID- 2567479 TI - Latest AIDS figures. PMID- 2567480 TI - Nicotine and cannabinoids as adjuncts to neuroleptics in the treatment of Tourette syndrome and other motor disorders. AB - Animal studies suggest nicotine and cannabinoids may significantly enhance the therapeutic value of neuroleptics in motor disorders. This was recently demonstrated in humans by the finding that chewing nicotine gum produced striking relief from tics and other symptoms of Tourette syndrome not controlled by neuroleptic treatment alone. It appears that the use of nicotine or cannabinoids may greatly improve the clinical response to neuroleptics in motor disorders. PMID- 2567481 TI - Nicotine induced excitation of locus coeruleus neurons is mediated via release of excitatory amino acids. AB - Previous electrophysiological studies have shown that systemically administered nicotine in low doses increases the firing rate of rat locus coeruleus (LC) neurons. In the present study, this action of nicotine was found to be prevented by pretreatment with kynurenic acid (1 mumol; i.c.v.). In addition, pretreatment with MK 801 effectively blocked the nicotine induced LC excitation in most neurons tested (60%) whereas the rest were left unaffected by this treatment. It is suggested that excitatory amino acids (EAA), e.g. released from nerve terminals from nucleus paragigantocellularis (PGi), indirectly mediated the effect of nicotine on LC neurons. PMID- 2567482 TI - In vivo demonstration of the enhancement of MK-801 by L-glutamate. AB - MK-801 infused bilaterally into the nucleus accumbens of rats produced a dose related increase in locomotor activity that was not blocked by intra-accumbens infusion of haloperidol (2.5 micrograms). Intra-accumbens infusion of L-glutamate (1.0 microgram) was without effect when administered alone, but significantly enhanced the increase in locomotor activity produced by MK-801 (5.0 micrograms). The inactive isomer of MK-801 did not produce hypermotility and L-glutamate did not enhance amphetamine(intra-accumbens)-induced hypermotility. These results extend to an in vivo model electrophysiological and radioligand binding studies demonstrating an enhancement of MK-801 activity by L-glutamate. The results also reinforce the concept that such an enhancement would occur during an escalation of excitatory amino acid levels accompanying pathological overload and, thus, would trigger a compensatory neuronal protection by MK-801. PMID- 2567483 TI - Alterations in plasma lipids consequent to endurance training and beta-blockade. AB - Alterations in plasma lipids consequent to endurance training and beta-blockade. Med. Sci. Sports Exerc., Vol. 21, No. 3, pp. 288-292, 1989. The chronic use of beta adrenergic blockers (BAB) has been associated with reductions in HDL cholesterol (HDL-C) and increases in triglycerides (TG). This study evaluated the impact of concurrent endurance exercise training and chronic medication with BAB on plasma lipid and lipoprotein profiles in healthy young adult males. Changes in plasma lipids and lipoproteins were investigated while exercise training under the influence of one of two nonselective BAB [sotalol (320 mg.d-1) and propranolol (160 mg.d-1)], one beta 1 selective BAB [atenolol (100 mg.d-1)], or a placebo control. Total cholesterol (TC), HDL-C, LDL-cholesterol (LDL-C), TG, and the ratios of TC/HDL and LDL/HDL were determined before and after endurance training programs of either 14 (N = 27, sotalol) or 15 (N = 47, propranolol/atenolol) wk duration. The subjects exhibited increases in maximal oxygen uptake of 12-20%. Despite increased endurance capacity, the subjects in both BAB and placebo control groups failed to demonstrate the expected increase in HDL-C and decrease in TG. In fact, HDL-C was significantly decreased post training in the propranolol group. The placebo groups did decrease TC, LDL-C and the TC/HDL and LDL/HDL ratios, improving their CHD risk profile. Similar changes were not observed in the groups on BAB. Thus, with respect to the present population, BAB does appear to interfere with the usual training-induced improvements in the lipid profile. Endurance training may, however, reduce the deterioration in the lipid profile known to occur with BAB. PMID- 2567484 TI - Maximal isokinetic cycle ergometry in patients with coronary artery disease. AB - We assessed the utility of short-term (30 s) maximal isokinetic cycle ergometry as an additional method of investigating the limitations to exercise in 33 carefully selected patients with documented coronary artery disease. The technique proved safe and reproducible in these patients. In relation to normal standards, performance was better in the maximal isokinetic cycle ergometer test (peak power = 819 +/- 116 W; average power = 532 +/- 72 W; total work = 13.1 +/- 2.1 kJ; 95-101% of predicted) than in the progressive incremental exercise test (VO2 = 1.80 +/- 0.37 l.min-1; power output = 919 +/- 165 kpm.min-1; 70-80% of predicted). Beta blockade did not affect maximal performance during either isokinetic or progressive incremental cycling, although maximal heart rate was significantly lower during both tests in patients on beta blockade. Power output in the progressive exercise was not as strongly related to the indices measured during the 30 s isokinetic test (r = 0.59-0.63) as it was in previous studies of healthy individuals (r = 0.89). The ability to detect individual variations in short-term exercise capacity measured with maximal isokinetic cycle ergometry may have significant potential value 1) as an additional method of determining the limitations to exercise and 2) when executing an exercise prescription in patients with coronary artery disease. PMID- 2567485 TI - Pili of Vibrio cholerae O1 biotype E1 Tor: a comparative study on adhesive and non-adhesive strains. AB - Pili were found on the cell surface of non-adhesive Vibrio cholerae O1 Biotype E1 Tor as well as the adhesive strain. Purified pili of the adhesive and non adhesive strains were morphologically, electrophoretically, and immunologically, indistinguishable from each other. The molecular weights of both pilin (subunit protein of the pilus) were about 16,000 daltons as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These 16 kDa pili are different from the pilus colonization factor, which is a 20.5 kDa protein, reported by Taylor et al. The 16 kDa pili of Vibrio cholerae O1 Biotype E1 Tor have hemagglutinating activity, but may have no role in colonization, because non-adhesive strains also have such pili. PMID- 2567486 TI - Influence of transglutaminase on the functions of mouse peritoneal macrophages. AB - Influence of transglutaminase on the production of interleukin-1 (IL-1) and on the release of active oxygen from mouse peritoneal macrophages was examined using cystamine and methylamine, an enzyme inhibitor and a substrate inhibitor, respectively. Casein-elicited or lipopolysaccharide (LPS)-elicited macrophages have higher levels of transglutaminase activity in comparison with resident macrophages, and there exists a definite correlation between endocytosis of erythrocytes and transglutaminase activity in either group of macrophages. The release of IL-1 by resident macrophages stimulated with LPS in vitro was significantly inhibited by the treatment with both transglutaminase inhibitors. However, these inhibitors were not able to inhibit the release of IL-1 from casein-elicited macrophages stimulated with LPS in vitro. The production of active oxygen from LPS-elicited macrophages was inhibited in a dose-dependent manner by the treatment of macrophages with cystamine, but was not by the treatment with methylamine. However, the treatment of LPS-elicited macrophages with cystamine did not inhibit the uptake of glucose into macrophages. These results suggest that transglutaminase activity in mouse peritoneal macrophages is an important factor for macrophage functions. PMID- 2567487 TI - Myocardial revascularization: a 20-year lesson. PMID- 2567488 TI - [Receptor diseases. 1: Disorders of adrenergic receptors, cholinergic receptors and neurotransmitters]. PMID- 2567489 TI - Drugs for rheumatoid arthritis. PMID- 2567490 TI - [Blood testosterone in cryptorchid newborn infants: does it have a prognostic significance?]. AB - Testosterone, LH, FSH, blood levels at birth were determined in 40 Italian true cryptorchid , 12 pseudo-cryptorchid and 71 healthy newborns. Testosterone concentrations were interpreted according to the localisation of the retained testicles. A greater incidence of spontaneous remission of the cryptorchidism was found in those subjects with: 1) testosterone plasma levels overlapping the neonatal range; 2) testicles placed in proximity to the external orifice of the inguinal tunnel. Follow-up of these infants up to 2 years of age will test the hypothesis. PMID- 2567491 TI - HLA antigens in drug-induced parkinsonism. AB - The results of two epidemiological studies suggest a hereditary predisposition to develop drug-induced parkinsonism. We investigated human leukocyte antigen (HLA) antigen prevalence rates in patients with neuroleptic-induced parkinsonism. Fifty two male, white, neuroleptic-treated, chronic in-patients with DSM-III-diagnosed schizophrenia were examined for the presence of parkinsonism. Subjects were tested for 23 type A, 43 type B, 4 type C, and 10 type DR HLA antigens. The group of schizophrenic patients with parkinsonism (n = 29) was compared with the group of schizophrenic patients without parkinsonism (n = 23). There were no significant differences between the two groups with respect to age, duration of neuroleptic exposure, or anticholinergic drug exposure. One HLA antigen, B44, was significantly more prevalent in the group with parkinsonism than in the group without parkinsonism. We derived a relative risk of 7.16 for drug-induced parkinsonism with HLA-B44 present in this group of schizophrenic patients. These data indicate that HLA-B44 may play a role in genetic or immunologic susceptibility to develop drug-induced parkinsonism in white schizophrenic individuals. PMID- 2567492 TI - Tardive akathisia: an analysis of clinical features and response to open therapeutic trials. AB - In recent years, there has been increasing recognition that akathisia occurs not only as an acute, self-limited complication of dopamine (DA) antagonist treatment, but also as a persistent form, called tardive akathisia. We represent a retrospective analysis of clinical features and therapeutic trials in 52 cases of this disorder. Although most patients developed this disorder after years of DA antagonist treatment (mean = 4.5 years), a significant proportion (34%) developed it within 1 year. The characteristic motor features included frequent, complex stereotyped movements. The legs were most frequently involved, showing marching in place and crossing/uncrossing. Trunk rocking, respiratory grunting and moaning, and complex hand movements such as face rubbing or scratching also occurred. In the 26 patients who were able to discontinue DA antagonists, akathisia persisted for years (mean = 2.7 years, range of 0.3-7 years) until abatement of symptoms or last follow-up. Younger patients were more likely to have remission or therapeutic suppression of akathisia at follow-up. In our experience, the catecholamine-depleting drugs reserpine and tetrabenazine were the most effective agents for suppressing symptoms, producing improvement in 87 and 58% of patients treated, respectively. However, improvement was limited in many patients, and at last follow-up only 33% of patients had complete abatement of their symptoms. In conclusion, tardive akathisia is a particularly disabling form of tardive dyskinesia, frequently persistent for years and often resistant to therapy. PMID- 2567493 TI - Cloning of a polymorphic DNA fragment from the genome of Leishmania donovani. AB - Recombinant DNA clones, containing highly repetitive DNA sequences, have been isolated from a Leishmania donovani genomic DNA library prepared in the replacement vector lambda gt.WES.lambda B. Two clones, probably telomeric in location, have been characterised and show a restriction fragment size polymorphism. Evidence is presented which suggests that L. donovani is diploid for this cloned genomic locus. PMID- 2567494 TI - Gene exchange in African trypanosomes: frequency and allelic segregation. AB - The existence of a system of genetic exchange in Trypanosoma brucei is now established, but the frequency with which mating occurs and the mechanisms by which genes are exchanged are still unknown. This paper presents the results of a study of one pair of trypanosome stocks, which show that mating is a non obligatory but frequent event in a life-cycle stage within the insect vector. Analysis of ten progeny clones using a total of eleven markers (iso-enzymes and DNA probes detecting restriction fragment length polymorphisms) has indicated that segregation of alleles occurs at five of these loci. The segregation patterns of a polymorphic EcoRI site in the maxi-circle of the kinetoplast DNA (kDNA) show that the progeny inherit one or other of the parental kDNA types. These results demonstrate that segregation of alleles occurs and that new combinations of alleles at different loci are generated in the progeny clones. The implications of these findings for defining the mechanism of gene exchange are discussed in relation to a simple mendelian genetic system involving meiosis and syngamy. PMID- 2567495 TI - AIDS conference. Just another meeting? PMID- 2567496 TI - DNA fingerprinting on trial. PMID- 2567497 TI - The CD2 antigen associates with the T-cell antigen receptor CD3 antigen complex on the surface of human T lymphocytes. AB - T lymphocytes can be activated by various stimuli directed either against the T cell antigen receptor-CD3 antigen complex (Ti-CD3) or the CD2 molecule; see ref. 1 for a review. Activation signals generated by antigen binding to the antigen specific alpha/beta heterodimer (Ti) are thought to be transduced via the invariant CD3 gamma, epsilon and delta chains, and the associated zeta and eta subunits. The physiological role of the interaction of CD2 with its homologous cell-surface associated ligand LFA-3 remains to be fully elucidated. It has been suggested that CD2 regulates an antigen-independent pathway of activation or that signals delivered via CD2 are an integral part of the antigen-specific pathway. Several recent studies have indicated a requirement for the Ti-CD3 complex in CD2 signalling. Thus, mutant T-cell lines expressing CD2, but not Ti-CD3, on the cell surface cannot be activated via the CD2 molecules. Functional interaction between the Ti-CD3 complex and the CD2 antigen suggests that these T-lymphocyte cell surface structures are physically associated. Here we use a digitonin-based solubilization procedure to explore this possibility and show that 40% of the cell-surface CD2 molecules can be specifically co-precipitated in association with the Ti-CD3 complex. PMID- 2567498 TI - Neu receptor dimerization. PMID- 2567499 TI - Case for diagnosis. Periarteritis nodosa. PMID- 2567500 TI - Two genes associated with liver cancer are regulated by different mechanisms in rasT24 transformed liver epithelial cells. AB - We compared the regulation of gamma-glutamyl transferase (gamma GT) and glutathione-S-transferase-P (GST-P) expression in rat liver epithelial cells (228 cells) and a line derived from them (C5 cells) by stable transfection with a metallothionein-activated ras fusion gene (MTrasT24). Earlier studies demonstrated that steady state RNA levels of these genes are increased after transformation of liver cells by MTrasT24 (Proc. Natl. Acad. Sci., 85, 344-348, 1988). In the present study, we found that the rate of gamma GT transcription increased approximately 20 fold after transformation by MTrasT24 while the rate of GST-P transcription increased no more than two fold. However, the stability of GST-P RNA was increased about 3 fold in these cells. Comparisons of gamma GT RNA stability were not possible since nontransformed liver cells (228) contain little or no gamma GT RNA. Thus, the accumulation of gamma GT RNA in C5 cells is heavily dependent on increased rates of transcription while the more modest increases in GST-P RNA levels result in large part from increased RNA stability. In ras transformed cells both transcriptional and post-transcriptional events contribute to the increased steady state RNA levels of cellular genes. PMID- 2567501 TI - The complete cDNA sequence for the premature form of growth hormone of the flounder Paralichthys olivaceus. PMID- 2567503 TI - A three codon insertion/deletion polymorphism in the signal peptide region of the human apolipoprotein B (APOB) gene directly typed by the polymerase chain reaction. PMID- 2567502 TI - Isolation and mapping of a polymorphic DNA sequence pH30 on chromosome 4[HGM provisional no. D4S139]. PMID- 2567504 TI - TaqI RFLP in human adenylate kinase-1 (AK1) gene region on chromosome 9. PMID- 2567505 TI - A common SacI polymorphism in the gene for the M1 subunit of ribonucleotide reductase (RRM1). PMID- 2567506 TI - Description of an HLA-DQA1 RFLP allele [DQ alpha 4] defining DQw4/DRw8-bearing haplotypes. PMID- 2567507 TI - An arbitrary single copy DNA sequence VC85 [D1S85] detects a 500 bp insertion/deletion polymorphism on chromosome 1. PMID- 2567508 TI - Detection of a 3 allele AvaII RFLP by a single copy anonymous DNA sequence VC75 [D7S404] localized to chromosome 7. PMID- 2567509 TI - An arbitrary single copy sequence VC64 [D1S86] detects a moderate frequency TaqI RFLP on chromosome 1]. PMID- 2567511 TI - A high frequency two allele TaqI RFLP detected by an anonymous sequence VC61 [D2S65] on chromosome 2. PMID- 2567510 TI - An arbitrary single copy human DNA sequence VC63 [D4S129] detects a TaqI RFLP on chromosome 4]. PMID- 2567513 TI - Au, Cu, Sn, Zn and Ag ion release from some Cd-free gold solders. AB - The aim of this study was to determine the release of Au, Cu, Sn, Zn, and Ag from five commercially available Cd-free gold solders. The electropotential system in modified Fusayma solution was used. The cycle process was repeated between +/- 250 mV for 24 hours. To study the ion release, samples of solution were analysed during the process. Rapid release of Zn from most alloys was found. Cu and Ag ions were also released, but only small amounts of the latter. Au showed very good passivation up to 24 hours. The use of soldered joints should be reduced in order to minimize corrosion. PMID- 2567512 TI - Morphological aspects of glucagon and somatostatin islet cells in diabetic bio breeding and low-dose streptozocin-treated Wistar rats. AB - We noted discrepancies in the results of differing investigators studying the fate of pancreatic A and D cells in two apparently comparable diabetic animal models, namely spontaneous diabetes occurring in the Bio Breeding (BB) rat and that induced by multiple low-dose treatment with streptozocin. Our aim was twofold: (a) to clear up these inconsistences in order to evaluate whether these two experimental models are truly comparable; and (b) to add further results regarding the ultrastructural aspects of A and D cells after onset of diabetes. We therefore observed the postdiabetic ultrastructural changes involving the cell population of the islet of Langerhans using the above mentioned models. Results showed that: (a) in BB rats, A cells do not undergo significant changes whereas D cells numbers decrease significantly; (b) in the low-dose streptozocin-treated Wistar rats A cells do not undergo significant changes whereas D cell numbers increase. Our observations provide evidence that changes affecting D cells in "spontaneous" diabetes differ from those observed in the "induced" type. PMID- 2567514 TI - The PapG adhesin of uropathogenic Escherichia coli contains separate regions for receptor binding and for the incorporation into the pilus. AB - Most uropathogenic strains of Escherichia coli produce heteropolymeric organelles, known as P pili, that bind to the globoseries of glycolipids present in the urinary tract. The formation of a P pilus is the result of a family of related proteins being coordinately assembled into the structure in a defined order with the adhesin located exclusively at the tip. The preassembled digalactoside alpha-D-galactopyranosyl-(1----4)-beta-D-galactopyranose-binding adhesin was purified to homogeneity from the periplasmic space in a complex with the periplasmic assembly protein PapD by affinity chromatography to alpha-D galactopyranosyl-(1----4)-beta-D-galactopyranose-Sepharose. A receptor-binding domain was mapped to the amino-terminal half of the adhesin. The interaction of PapD with PapG, which was required for the incorporation of the adhesin into the pilus, was found to protect PapG from proteolytic cleavages and enhanced the processing of the PapG signal peptide. A preassembly domain necessary for forming a complex with PapD was mapped to the carboxyl terminus of PapG. PMID- 2567515 TI - A region in the coding sequence is required for high-level expression of murine histone H3 gene. AB - Replication-dependent histone genes are expressed at high rates in S phase to provide the histone proteins required for chromosomal replication. Two genes, an H2a and H3 gene, located on chromosome 3 in the mouse and cloned together in a single 3-kilobase (plasmid MM614) restriction fragment are highly expressed. By transfecting mouse histone gene constructs into Chinese hamster ovary cells, we have identified a 110-nucleotide region within the coding sequence of the H3.2 614 gene that is required for high-level expression. Deletion of this region reduces expression of the gene by 20-fold. Additionally, the histone-coding region activates the human alpha-globin promoter, which is normally not expressed well in Chinese hamster ovary cells. Similar results with deletion constructions involving the H2a-614 gene suggest that an intragenic region plays an important role in transcription of these genes. PMID- 2567516 TI - Clonal contributions of small numbers of retrovirally marked hematopoietic stem cells engrafted in unirradiated neonatal W/Wv mice. AB - Mice were repopulated with small numbers of retrovirally marked hematopoietic cells operationally definable as totipotent hematopoietic stem cells, without engraftment of cells at later stages of hematopoiesis, in order to facilitate analysis of stem cell clonal histories. This result depended upon the use of unirradiated W/Wv newborn recipients. Before transplantation, viral integration markers were introduced during cocultivation of fetal liver or bone marrow cells with helper cell lines exporting defective recombinant murine retroviruses of the HHAM series. Omission of selection in culture [although the vector contained the bacterial neomycin-resistance (neo) gene] also limited the proportion of stem cells that were virally labeled. Under these conditions, engraftment was restricted to a small population of marked and unmarked normal donor stem cells, due to their competitive advantage over the corresponding defective cells of the mutant hosts. A relatively simple and coherent pattern emerged, of one or a few virally marked clones, in contrast to previous studies. In order to establish the totipotent hematopoietic stem cell identity of the engrafted cells, tissues were sampled for viral and inbred-strain markers for periods close to one year after transplantation. The virally labeled clones were characterized as stem cell clones by their extensive self-renewal and by formation of the wide range of myeloid and lymphoid lineages tested. Results clearly documented concurrent contributions of cohorts of stem cells to hematopoiesis. A given stem cell can increase or decrease its proliferative activity, become completely inactive or lost, or become active after a long latent period. The contribution of a single clone present in a particular lineage was usually between 5% and 20%. PMID- 2567517 TI - Mammalian mating systems. AB - Male mammals show a diverse array of mating bonds, including obligate monogamy, unimale and group polygyny and promiscuity. These are associated with a wide variety of different forms of mate guarding, including the defence of feeding and mating territories, the defence of female groups and the defence of individual receptive females. Female mating bonds include long-term monogamy, serial monogamy, polyandry and promiscuity. Both male and female mating behaviour varies widely within species. Variation in male mating behaviour is related to the effect of male assistance in rearing young and to the defensibility of females by males. The latter is, in turn, related to female ranging behaviour and to the size and stability of female groups. Much of the variation in mammalian mating bonds and systems of mate guarding can be attributed to differences in these three variables. PMID- 2567518 TI - Role of excitatory amino acid receptors in synaptic transmission in area CA1 of rat hippocampus. AB - The new antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), which blocks responses to kainate and quisqualate, has been used in conjunction with D-2-amino 5-phosphonovalerate (APV), which blocks selectively responses to N-methyl-D aspartate (NMDA), to determine the role of excitatory amino acid receptors in synaptic transmission. An excitatory postsynaptic potential (EPSP)-inhibitory postsynaptic potential (IPSP) sequence was evoked in CA1 neurons by stimulation of the Schaffer collateral-commissural pathway in rat hippocampal slices. CNQX (10 microM) substantially reduced the EPSP without having any effect on input resistance or membrane potential. The IPSP was also reduced provided that the stimulating electrode was place approximately 1 mm from the recording electrode. The EPSP that remained in the presence of CNQX had characteristics of an NMDA receptor-mediated potential; it had a slow timecourse, summated at high frequencies, was blocked reversibly by APV, increased greatly in size in Mg2+ free medium, and showed an anomalous voltage dependence in Mg2+-containing medium. In the presence of CNQX, an APV-sensitive polysynaptic GABAergic IPSP could be evoked, indicating that NMDA receptors can mediate suprathreshold EPSPS in inhibitory interneurons. It is suggested that either NMDA or non-NMDA receptors can, under different circumstances, mediate the synaptic excitation of pyramidal neurons and inhibitory interneurons in area CA1 of the hippocampus. PMID- 2567519 TI - Ocular vergence under natural conditions. I. Continuous changes of target distance along the median plane. AB - Horizontal binocular eye movements of four subjects were recorded with the scleral sensor coil--revolving magnetic field technique while they fixated a natural target, whose distance was varied in a normally illuminated room. The distance of the target relative to the head of the subject was changed in three ways: (a) the target was moved manually by the experimenter; (b) the target was moved manually by the subject; (c) the target remained stationary while the subject moved his upper torso towards and away from the target. The rate of change of target distance was varied systematically in four levels, ranging from 'slow' to 'very fast', corresponding to changes in target vergence from about 10 degrees s-1 to about 100 degrees s-1. The dynamics of ocular vergence with regard to delay and speed were, under all three conditions, considerably better than could be expected from the literature on ocular vergence induced by disparity and/or blur. When 'very fast' changes in the distance of the target were made, subjects achieved maximum vergence speeds of up to about 100 degrees s-1. Delays of these fast vergence responses were generally smaller than 125 ms. Negative delays, i.e. ocular vergence leading the change in target distance, were observed. The eyes led the target (i.e. predicted target motion) by about 90 ms on average, when the subject used his hand to move the target. Vergence tracking was almost perfect when changes in distance were produced by moving the upper torso. In this condition, the eye led the target by about 5 ms. In the 'slow' and 'medium' conditions (stimulus speeds about 10-40 degrees s-1) tracking was accurate to within 1-2 degrees, irrespective of the way in which the target was moved. In the 'fast' and 'very fast' conditions (stimulus speeds about 40-100 degrees s-1), the accuracy of vergence tracking was better for self-induced than for experimenter-induced target displacements, and accuracy was best during voluntary movements of the upper torso. In the last case, ocular vergence speed was within about 10% of the rate of change of the vergence angle formed by the eyes and the stationary target. The dynamics of convergent and divergent vergence responses varied considerably. These variations were idiosyncratic. They were consistent within, but not between, subjects. Ocular vergence associated with attempted fixation of an imagined target, changing distance in darkness, could only be made by two of the four subjects.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2567520 TI - Ocular vergence under natural conditions. II. Gaze shifts between real targets differing in distance and direction. AB - Horizontal binocular eye movements of three subjects were recorded with the scleral sensor coil--revolving magnetic field technique during voluntary shifts of gaze between pairs of stationary, real, continuously visible targets. The target pairs were located either along the median plane (requiring symmetrical vergence), or on either side of the median plane (requiring asymmetrical vergence). Symmetrical vergence was primarily smooth, but it was often assisted by small, disjunctive saccades. Peak vergence speeds were very high; they increased from about 50 degrees s-1 for vergence changes of 5 degrees to between 150 and 200 degrees s-1 for vergence changes of 34 degrees. Differences between convergence and divergence were idiosyncratic. Asymmetrical vergence, requiring a vergence of 11 degrees combined with a version of 45 degrees, was largely saccadic. Unequal saccades mediated virtually all (95%) of the vergence required in the divergent direction, whereas 75% of the vergence required in the convergent direction was mediated by unequal saccades, with the remaining convergence mediated by smooth vergence, following completion of the saccades. Peak divergence speeds during these saccades were very high (180 degrees s-1 for a change of vergence of 11 degrees); much faster than the smooth, symmetrical vergence change of comparable size (14 degrees). Peak convergent saccadic speeds were about 20% lower. This difference in peak speed was caused by an initial, transient divergence, observed at the beginning of all horizontal saccades. The waveform of disjunctive saccades did not have the same shape as the waveform of conjugate saccades of similar size. The smaller saccade of the disjunctive pair was stretched out in time so as to have the same duration as its larger, companion saccade. These results permitted the conclusion that the subsystems controlling saccades and vergence are not independent. Vergence responses were relatively slow and incomplete with monocular viewing, which excluded disparity as a cue. Monocularly stimulated vergence decreased as a function of the increasing presbyopia of our three subjects. Subjects were able to generate some vergence in darkness towards previously seen and remembered targets. Such responses, however, were slow, irregular and evanescent. In conclusion, vergence shifts between targets, which provided all natural cues to distance, were fast and accurate; they appeared adequate to provide effective binocular vision under natural conditions. This result could not have been expected on the basis of previous observations, all of which had been made with severely reduced cues to depth.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2567521 TI - Hypothermia in mice: D2 dopamine receptor mediation and absence of spare receptors. AB - The nonselective dopamine (DA) receptor agonists R(-)apomorphine (APO) and R(-)-N n-propylnorapomorphine (NPA) elicited dose- and time-dependent hypothermia in mice with ED50 values of 300 and 18 micrograms/kg, respectively. The selective D2 agonist quinpirole (LY 171555) also elicited dose-dependent hypothermia, whereas the selective D1 agonist SKF 38393 had no effect. The selective D1 and D2 antagonists SCH 23390 (1 mg/kg) and sulpiride (200 mg/kg), respectively, did not significantly alter body temperature. The hypothermic effect of a maximal dose of NPA (0.2 mg/kg) was not blocked by SCH 23390 (1 mg/kg) but was significantly attenuated (p less than 0.001) by pretreatment with sulpiride (200 mg/kg). Pretreatment with sulpiride (200 mg/kg) produced a parallel, 40-fold shift to the right of the dose-response curve for NPA. Partial, irreversible DA receptor inactivation by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) (2 mg/kg) reduced the maximal hypothermic effect of NPA (to 49% of control) without altering its ED50. Analysis of the data indicated a linear relationship between DA receptor occupancy and hypothermic response. The results demonstrate that DA agonist-induced hypothermia in mice is mediated by D2 receptors and that there is no receptor reserved for this response. PMID- 2567523 TI - Amnesia of a passive avoidance task due to the beta 2-adrenoceptor antagonist ICI 118,551. AB - The selective beta 2-adrenoceptor antagonist ICI 118,551 induces amnesia in the domestic chick when given systemically, 10 min after a one-trial PAL task. Young chicks will spontaneously peck at a small bright bead. If the bead has been coated with a distasteful substance, the chicks will learn in a single trial not to peck at a similar bead on subsequent presentation. Administration of ICI 118,551 prevented retention of this task. Vehicle-injected chicks which learnt the task, avoided a similar bead to the training bead in the retention test, but did not avoid a bead of a different colour. The effect of ICI 118,551 is unlikely to be a direct effect on performance since amnesic chicks pecked both beads freely and equally in the test. PMID- 2567522 TI - Yohimbine's anxiogenic action: evidence for noradrenergic and dopaminergic sites. AB - Yohimbine (2.5 or 4 mg/kg) reduced the percentage of open arm entries and the percentage of time spent on the open arms displayed by rats on an elevated plus maze indicating anxiogenic activity. These effects were reversed by the alpha 2 adrenoceptor agonist clonidine (0.01 mg/kg) and by the dopamine receptor agonist apomorphine (0.57 mg/kg). The following failed to reverse the effects of yohimbine: the selective alpha 2-adrenoceptor agonists, guanfacine (0.25 and 1 mg/kg), B-HT920 (0.025 and 0.1 mg/kg), B-HT933 (1 and 10 mg/kg); the beta-blocker propranolol (2.5 and 10 mg/kg); the alpha 1-adrenoceptor agonist phenylephrine; the D1 agonist SK&F 38393 (5 and 10 mg/kg) and the D2 agonist LY 171555 (0.5 and 1 mg/kg). Therefore, it is unlikely that activity at only the alpha 1, alpha 2, beta, D1 or D2 sites can entirely account for the anxiogenic actions of yohimbine in the elevated plus-maze. Evidence that clonidine affects the dopaminergic system and that apomorphine affects the noradrenergic system suggest that yohimbine may produce its anxiogenic response by activity on both the noradrenergic and dopaminergic systems. PMID- 2567525 TI - An hypothesis regarding the antipsychotic effect of neuroleptic drugs. AB - The antipsychotic effect of neuroleptic drugs appears gradually over the course of several weeks of chronic drug administration. Neuroleptic drugs are thought to act by blocking dopamine receptors; however, the dopamine-blocking effect of neuroleptics appears rapidly. One effect of dopamine antagonists which develops slowly is dopaminergic supersensitivity. It is suggested that this dopaminergic supersensitivity is related to the development of tolerance to some of the acute sedative properties of neuroleptics, but not to the antipsychotic effect. A population of glutamate receptors which are postsynaptic to the cortico-striatal afferents is located on the same neurons as striatal dopamine receptors. These glutamate synapses are located on the heads of dendritic spines of striato-nigral projection neurons, while the dopaminergic synapses are predominantly located on the necks of these same dendritic spines. Similar relationships could exist for mesolimbic and mesocortical dopamine systems. In peripheral systems, postjunctional denervation supersensitivity is known to be nonspecific; in other words, denervation of a single innervation of an excitable cell can alter the response to a range of stimuli. The antipsychotic effect of neuroleptics is therefore suggested to be due to nonspecific postjunctional subsensitivity at glutamate synapses, which develops concomitant with supersensitivity at dopaminergic synapses. PMID- 2567524 TI - Effects of 5-HT1A receptor agonists and L-5-HTP in Montgomery's conflict test. AB - The effects of the pyrimidinyl-piperazines buspirone, gepirone, ipsapirone and their common metabolite 1-(2-pyrimidinyl)-piperazine (PmP) as well as of 8 hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) and L-5-hydroxytryptophan (L-5 HTP) were investigated in Montgomery's conflict test--an animal anxiety model based on the animal's inborn urge to explore a new environment and its simultaneous fear of elevated, open spaces. Subcutaneous buspirone (32-128 nmol/kg), gepirone (32-128 nmol/kg), ipsapirone (32-512 nmol/kg) and 8-OH-DPAT (50-200 nmol/kg), as well as intraperitoneal L-5-HTP (56 mumol/kg) produced anxiolytic-like effects. However, at higher doses the magnitude of these effects decreased and overall the dose-response curves displayed inverted U-shapes. The highest doses (2048 nmol/kg) of buspirone and of gepirone even decreased responding below control levels, possibly in part due to concomitant sedation/motor impairment. After L-5-HTP (448 mumol/kg) and PmP (512 nmol/kg) anxiogenic-like effects were observed. The results indicate that anxiolytic- and anxiogenic-like effects of drugs affecting central serotonergic neurotransmission can be obtained in a sensitive rat anxiety model which neither involves consummatory behavior nor punishment. The anxiolytic-like effects of these compounds may be due to their 5-HT1A agonistic properties. Moreover, the present data may provide support for a possible reciprocal association of presynaptic 5 HT1A receptors vs. postsynaptic 5-HT1A as well as 5-HT2 receptors with regard to anxiety. PMID- 2567526 TI - Functional supersensitivity of the hippocampal dopaminergic system after prolonged treatment with haloperidol. AB - The effect of acute and prolonged (21 days) treatment with haloperidol (1 or 5 mg/kg SC) on the dopamine-, apomorphine- and LY 171555-induced changes in the firing rate of CA1 layer neurons was studied in a hippocampal slice preparation. Dopamine and apomorphine administration evoked either an excitatory or inhibitory reaction, while the selective D2 receptor agonist LY 171555 increased the firing rate of hippocampal neurons. The excitatory effects of dopamine and LY 171555 were blocked by sulpiride and haloperidol. Prolonged administration of haloperidol potentiated the excitatory reaction of dopamine and apomorphine; however, even a single dose of the neuroleptic enhanced the dopamine-induced effect. The reaction evoked by LY 171555 was not significantly affected by either acute or chronic treatment with haloperidol. The present findings indicate that long-term administration of haloperidol results in an increased sensitivity of hippocampal neurons to the mixed dopamine agonists, dopamine and apomorphine, but not to selective stimulation of the D2 receptor. PMID- 2567527 TI - Neurochemical and cognitive aspects of anxiety disorders. PMID- 2567528 TI - Drug effects on active immobility responses: what they tell us about neurotransmitter systems and motor functions. AB - The literature reviewed indicates that active immobility can be promoted by systemic injections of various neurotransmitter systems, as follows: (1) Dopaminergic blockade of both D1 and D2 receptor subtypes. (2) Cholinergic agonism of both muscarinic and nicotinic receptors. (3) Noradrenergic agonism of both alpha-1 and alpha-2 receptors (but these agonists may interfere with haloperidol- and reserpine-induced catalepsy). (4) GABA agonism. (5) Histamine agonism, particularly at the H1 receptor. (6) Opiate agonism, including action of many endogenous opiate peptides, particularly those affecting mu and delta receptors. (7) Agonism by certain other peptides (neurotensin, cholecystokinin). Among the major interactions of neurotransmitter systems that regulate immobility, are the following: (1) Cholinergic-dopaminergic (cholinolytics disrupt catalepsy of dopaminergic blockade and dopaminergic agonists tend to disrupt cholinomimetic catalepsy). (2) Opiate-induced catalepsy is antagonized by the dopamine agonist, apomorphine, but is enhanced by amphetamine. It is also antagonized by certain alpha-2 adrenergic agonists, while it does not seem to be antagonized by anticholinergics. (3) Numerous other interactions have been reported, involving opiates and MSH, serotonin and dopamine mimetics, serotonin and ketamine, GABA and neuroleptics, neurotensin and anticholinergics and histamine. The significance of the multiple neurotransmitter systems is unknown. One possible explanation is that the various neurotransmitter systems participate in mediating the sensory inputs that are involved in triggering immobility and regulate the higher-order limbic and basal ganglia processing reactions that engage a final motor output pathway from the brainstem. The brain is assumed to contain two sets of systems, each with its own, or possibly overlapping, set of neurotransmitter systems, that promote either active immobility or locomotion. The systems reciprocally inhibit each other. Another view, not mutually exclusive, is that output from the locomotor-promoting system provides a negative feedback, via the active immobility pathways, to act as a "brake" on movement, while at the same time maintaining the muscular tonus that is characteristic of active immobility. PMID- 2567529 TI - [Beta blockaders and portal hypertension]. PMID- 2567530 TI - [Proceedings of an international seminar "Xenograft 25". Renaissance of xenogene transplantation (New York, 11-13 November 1988)]. PMID- 2567532 TI - Hecklers and protesters liven up a dull meeting. PMID- 2567531 TI - Restriction fragment length polymorphism analysis of T-cell receptor genes in inflammatory bowel disease. AB - The etiology of inflammatory bowel disease is still unknown, but autoimmune phenomena are thought to play an important role. However, only a weak association between HLA or immunoglobulin allotypes and inflammatory bowel diseases has been noted. Recently, DNA markers (restriction fragment length polymorphism (RFLP) pattern) of T-cell receptor gene allotypes have been reported and shown to be linked to susceptibility to autoimmune disease. We investigated the T-cell receptor RFLP pattern in inflammatory bowel diseases. No linkage to the constant region alpha- and beta-chain markers was observed, and no differences in the frequencies of 'genoallotypes' was found between patients and normal blood donors. PMID- 2567533 TI - Transfer RNA genes: landmarks for integration of mobile genetic elements in Dictyostelium discoideum. AB - In prokaryotes and eukaryotes mobile genetic elements frequently disrupt the highly conservative structures of chromosomes, which are responsible for storage of genetic information. The factors determining the site for integration of such elements are still unknown. Transfer RNA (tRNA) genes are associated in a highly significant manner with different putative mobile genetic elements in the cellular slime mold Dictyostelium discoideum. These results suggest that tRNA genes in D. discoideum, and probably tRNA genes generally in lower eukaryotes, may function as genomic landmarks for the integration of different transposable elements in a strictly position-specific manner. PMID- 2567534 TI - Multiple stings by imported fire ants (Solenopsis invicta), without systemic effects. AB - This is the first case report of imported fire ants invading a building and aggressively attacking a human being, resulting in multiple stings. This case illustrates that, although the venom has a high LD50 in mice, it does not induce toxic systemic effects in humans when the venom is introduced intradermally. PMID- 2567535 TI - [Endocrine cells of the gastroduodenal area in duodenal ulcer]. AB - A study has been carried out to substantiate the assumption that excessive proliferation of major gastroduodenal endocrine cells may be predetermined genetically. Hereditary predisposition to the hyperplasia of cells belonging to the APUD system may be inherited both through the male and female lines. Duodenal ulcers (DU) undoubtedly involves multifactorial inheritance. Acquired capacity for endocrine cell hyperplasia cannot be excluded in some of DU patients. The development of DU is, apparently, related to the formation of a pathological system, based on pathologic structure-linked neurohormonal relationships between the duodenum and peripheral and central nervous systems that enhance nerve impulses in the duodenal bulbar portion. A sudden "total" catecholamine release occurs at a certain stage and is followed by a change of production/utilization ratios of various hormones that weakens pathologic neurohormonal relationships, destroys the pathologic system and activates the healing process around the ulcer. The pathologic system, active in peptic ulcers, is doubtless counteracted by a system, whose activity must be determined by the number of endocrine cells with beta-endorphine-like and, perhaps, serotonin-like immunoreactivity. The antagonistic regulation principle is of great universal significance for general biology. It is essential for healing and chronization processes. PMID- 2567536 TI - [Diagnostic difficulties in periarteritis nodosa]. PMID- 2567537 TI - [Changes of plasma and tissue gamma-glutamyltransferase under the influence of drugs]. AB - Measurement of gamma-glutamyltransferase (GGT) activity in plasma is widely used in clinical biology (in order to detect hepatic diseases or to monitor treatment for alcoholism), and also in pharmacology (since this test is the only plasmatic marker for hepatic induction in human). However, the correct interpretation of a high plasmatic activity should take into account the various analytical factors which can affect results, as well as the physiological parameters known to modify this activity. It also requires its comparison to defined reference values. Several mechanisms may be involved in the increase of plasmatic activity as an index of hepatic induction, such as an increase in the protein synthesis, a release of the enzyme from the membrane or a modification in the biliary flux. PMID- 2567538 TI - [Development of atherosclerosis is not influenced by beta blockaders]. PMID- 2567539 TI - Definition of DRw10 by restriction fragment length polymorphism. AB - To better define the presence of the DRw10 haplotype which has sometimes proved difficult to type by using serologic reagents, Southern blot analysis was performed on seven DRw10 heterozygous individuals with rheumatoid arthritis. Using the restriction enzymes Taq I or BamH I, the restriction fragment length polymorphism (RFLP) pattern for the DRw10 haplotype was clearly distinguishable from that of other DR alleles. Digestion with Taq I revealed a unique DR beta/Taq I 12.20 fragment. A characteristic DR beta/Taq I 4.60 fragment was also present only in DRw10 and DR1 haplotypes. Digestion with the restriction enzyme BamH I revealed a DR beta/BamH I 5.07 fragment also present in DRw10 and DR1 haplotypes, and a DR beta/BamH I 4.30 fragment shared with the DRw52 and DR2 haplotypes but not found in DR1 haplotypes. The pattern was readily distinguished from those given by the haplotypes DR4, 7 and w9. Family studies of five individuals demonstrated appropriate segregation of the restriction fragments. In particular, segregation of DRw10 haplotypes from DR1 haplotypes was clearly shown in a family in which the DRw10 haplotype was associated with rheumatoid arthritis in two individuals. Southern blot analysis proved to be a useful alternative method for identifying the DRw10 allele in certain combinations where this allele has been difficult to define serologically. PMID- 2567541 TI - Blocking of cyclosporine immunosuppression by neuroleptics. PMID- 2567542 TI - [Histamine-2 blockade in skin disease]. PMID- 2567540 TI - Evidence that class I-restricted response to Thy-1 antigen requires L3T4+ cells and macrophages but not Lyt-2+ cells. PMID- 2567543 TI - [Treatment of acute myocardial infarction--an elucidative report]. AB - The present-day optimal treatment of patients with acute myocardial infarction (AMI) is reviewed. The prehospital phase should be as brief as possible. Emergency observation and treatment in hospital should be initiated without delay. Schematic stages for mobilization have been discarded and free mobilization is recommended. Routine acute intervention with thrombolysis is recommended for patients in whom symptoms have been present for 6-12 hours and treatment with Aspirin is recommended. Beta-blocking agents are recommended for patients with increased risk after discharge. Treatment of ventricular and supraventricular arrhythmias, block and cardiac failure are reviewed in detail. Patients without complications should be monitored for three to five days and may be discharged after seven to ten days. Exercise ECG should be carried out at discharge to assess the working capacity, ischaemia and subjective reaction. The importance of good patient information is emphasized. Cessation of smoking, control of lipids and blood pressure are important as secondary interventions. As far as possible, outpatient control should be offered after discharge. The criteria for referral to specialized cardiological departments are established both for emergency and elective referral. Patients under the age of 70 years with high risk for repeated AMI or death after discharge (with residual ischaemia) should possibly be referred for coronary arteriography. PMID- 2567544 TI - [The normal values of gamma-glutamyltransferase are falsely defined up to now: on the diagnosis of hypertension, obesity and diabetes with reference to "normal" consumption of alcohol]. AB - In a large sample of 1379 adult patients and, in addition, in a smaller group of 223 other patients in whom a glucose tolerance test with measurement of serum insulin was carried out, an increase of blood pressure, pulse rate, relative body weight and serum insulin was found which correlated significantly with that range of gamma glutamyltransferase (GGT) values which erroneously so far is considered to be normal. The really normal range of the GGT is not up to 28 (measured at 25 degrees C), but only up to 10 U/l. Persons with GGT 9-12 U/l have a significantly higher blood pressure than persons with GGT up to 8 U/l. The relationship between blood pressure and GGT is the same in males and females although the females show a higher GGT for the same amount of alcohol consumed; in both, males and females, the steepest increase is just in the low GGT range between 9 and 25 U/l. The nature of this ethanol-effect is toxic, not caloric. Daily alcohol in "normal" ("social") amounts causes hyperinsulinemia (and thus increased sodium reabsorption in the Kidney) as well as increased catecholamine excretion. "Normal" alcohol consumption leading to hepatic steatosis as the "normal" condition of the population, has more health hazards than so for assumed. A GGT higher than 10 U/l (measured at 25 degrees C), is besides hyperinsulinemia the most sensitive test for pathologic changes of the metabolism and the cardiovascular parameters due to hepatic steatosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567545 TI - Lymphocyte mediated cell lysis. AB - Lymphocyte-mediated cell lysis represents an important immunologic effector mechanism involved in defense against viral infections, allograft rejection, and tumor surveillance. Moreover, regulatory T cell interactions within the immune system are based, at least in part, on molecular events related to this function. The multiplicity of effector cell populations that can mediate cytotoxicity, the cell/cell interaction determinants which they require for execution of their activities, and molecular events underlying the lytic process itself, as elucidated recently, are the subjects of the present review article. PMID- 2567546 TI - The use of electron microscopy and immunocytochemistry to characterise spontaneously-arising, transplantable rat tumors. AB - When examined by light microscopy, transplanted animal tumors frequently bear little resemblance to the original neoplasm. If such tumors are to be used as models of human cancer they should be characterised as regards extant rather than historical features. Consequently, we have examined, by electron microscopy and immunocytochemistry, five spontaneously arising tumors transplantable in the WAB/Not rat that are currently diagnosed on the basis of historical features only. A typical sarcoma was used for comparison. Of four spontaneously arising tumors previously classified as carcinoma, Sp4 possessed epithelial features on both ultrastructural and immunocytochemical analysis, Sp107 on ultrastructural analysis only and Sp15 and Sp22 by neither technique. Expression of vimentin was most marked with Sp15 and Sp107. The putative sarcoma, Sp24, showed clear evidence of epithelial differentiation but no evidence of vimentin expression. This study (a) records the phenotypic drift of experimental tumors on transplantation (most clearly with Sp107) and the co-expression of cytokeratins and vimentin in putative carcinomas, (b) confirms the inadequacy of routine histology for accurate characterisation of such tumors and (c) details techniques for a more thorough assessment of state of differentiation that should guide the choice of experimental model. PMID- 2567547 TI - Cell cycle of normal bladder urothelium in developing and adult mice. AB - The present research has employed a novel, nonradioactive technique to quantitatively study normal urothelial proliferation in foetal, neonatal, juvenile and adult mouse bladder. Using whole mount histological preparations, the total number of urothelial nuclei per mouse bladder, and per given urothelial cell layer, have been assessed to provide data of the (unstimulated) kinetic behaviour of basal urothelial cells (the proliferative population), to analyse characteristics of the normal urothelial cell cycle. The urothelial cell cycle time increases from 30.6 h (foetal) to 40 weeks (adult), the duration of mitosis from 0.23 h (foetal) to 2.71 h (adult) and the duration of DNA synthesis from 2.52 h (neonatal) to 10.83 h (adult). These are average values for the urothelial cell cycle, which do not preclude the possible existence of proliferative units. The ratio of superficial nuclei to basal and intermediate nuclei, possibly indicative of a urothelial proliferative unit, declines to reach a plateau (1:40) in adult mice. These findings indicate that rapid urothelial proliferation during early murine development was likely to be a) biologically useful, since intrauterine foetal metabolic activity may require a functional bladder urothelium at an early stage, b) kinetically similar to acutely regenerating adult urothelial cells after cytotoxic insult. During murine life, the range of durations of mitosis and DNA synthesis is much less than the range of cell cycle times. Normal unstimulated urothelium of adult mice was confirmed to proliferate slowly. PMID- 2567548 TI - Spontaneous immune complex orchitis in brown Norway rats. AB - Immune complexes occur spontaneously in the testis of Brown-Norway (BN) inbred rats between the basal lamina of the seminiferous tubules and the outer lamina of the myoid testicular cells. The deposits can be detected immunohistologically (IgG; C3) and by electron microscopy. The immune complexes appear between the 8th and 12th weeks of life, increase in amount up to the 30th week and decrease thereafter. After about the 20th week, of life, 15% of the animals show destruction of the germinal epithelium accompanied by an infiltration of lymphocytes and plasma cells. The final stage of this disease, which initially shows no signs of inflammation, is characterized by diffuse tubular atrophy. However, up to the 70th week of life, 85% of the animals with immune complexes show no pathological alterations. Antibodies eluated from the testes react with spermatocytes I and structures close to the lumen of the seminiferous tubules, but not with mature sperms. Serum antibodies to sperms occur in about 25% of the BN rats, but the presence of these antibodies shows no correlation with the immunohistological findings. This newly described spontaneous immune complex orchitis is regarded as a further example of an in-situ-induced immune complex disease. The observations made here can be compared with those in (peri-) membraneous glomerulonephritis, another example of a disorder resulting from in situ-formation of immune deposits. PMID- 2567549 TI - Bone marrow analysis of the myelodysplastic syndromes: histological and immunohistochemical features related to the evolution of overt leukemia. AB - Bone marrow trephines from 31 patients with an initial diagnosis of myelodysplastic syndromes (MDS) were examined and analyzed histologically and immunohistochemically. In those cases terminating in overt leukemia (6/31, 19%), the number of bone marrow mast cells was significantly reduced, compared with those which did not evolve to overt leukemia. The bone marrow lymphoid cells that may participate in immunosurveillance against the proliferation of blast cells were also significantly reduced in cases terminating in overt leukemia. However, S-100 protein-positive cells, which include histiocytes and suppressor T-cells, were increased in cases terminating in overt leukemia. The results indicated that examination of the bone marrow to determine the proportions of mast cells and lymphoid cells which may be involved in host defense systems may be useful in predicting the evolution to overt leukemia in MDS. In the present series, patients with a hypocellular marrow (5/31, 16%) did not progress to overt leukemia and had a significantly lower bone marrow reticulin content, a significantly lower megakaryocyte count, a relatively higher mast cell count and a significantly higher lymphoid cell count than those with a normocellular or hypercellular marrow. These findings may reflect the initial features of MDS or, possibly, that hypocellular MDS is an independent entity with a low potential for blastic proliferation. PMID- 2567551 TI - Immunohistochemical localization of the alpha and beta subunits of S-100 protein in pleomorphic adenoma of the salivary glands. AB - The immunohistochemical expression of the alpha and beta subunits of S-100 protein in reactive, modified and transformed of myoepithelial cells, salivary pleomorphic was investigated using monoclonal antibodies. With S-100 alpha, normal salivary glands showed strong staining in serous acinar cells and moderate to slight staining in ductal segments, and with S-100 beta staining was slight or negative in acinar cells, but strong in nerve fibres. In pleomorphic salivary adenomas, the immunohistochemical distribution of S-100 alpha and beta proteins indicated great variation in the tumour cells. Some neoplastic cells gave similar staining for both S-100 alpha and beta, others were strongly positive for S-100 alpha and stained only slightly for S-100 beta, or vice versa. Yet other cells were positive for S-100 alpha and negative for S-100 beta, or vice versa. Pleomorphic salivary adenomas were classified both by histopathological criteria and by their staining pattern for S-100 alpha and beta proteins. Great heterogeneity in S-100 alpha and beta protein expression was found in individual tumour cells of both ductal and myoepithelial origin, and no regular pattern was identified. The cellular origin of salivary pleomorphic adenomas is discussed in terms of S-100 alpha and beta protein immunohistochemistry. Pleomorphic adenoma cells may be transformed from reserve cells into tumour cells displaying biologic properties of myoepithelial cells, ductal cells, or a mixture of both. PMID- 2567550 TI - Alpha smooth muscle actin (alpha-SM actin) in normal human ovaries, in ovarian stromal hyperplasia and in ovarian neoplasms. AB - An immunohistochemical investigation of alpha-smooth muscle actin (alpha-SM actin) using the monoclonal anti-alpha-SM-1 antibody was carried out in 15 normal ovaries, in three ovaries with stromal hyperplasia and in 27 neoplastic ovaries. In selected cases the pattern of actin isoforms was examined by means of 2 D-gel electrophoresis. In addition, the tissues were stained for vimentin and desmin. In normal ovaries alpha-SM actin was found in the inner cortex and in the theca externa. In ovarian stromal hyperplasia expression of alpha-SM actin was minimal or absent. In primary and metastatic epithelial tumors there was positive stromal staining for alpha-SM actin, especially in the vicinity of epithelial elements. This tended to be more widespread in malignant neoplasms. Thecomas did not express alpha-SM-actin and could thus be differentiated from leiomyomas which stained intensely for alpha-SM actin. Only focal stromal staining of alpha-SM actin was observed in granulosa and germ cell tumors. In all the tissues studied blood vessels were strongly positive for alpha-SM actin. Desmin, although present in the stroma of most of the specimens, was less abundant than alpha-SM actin. We concluded that alpha-SM actin is a component of the normal human ovary where it may contribute to the contractility of its stroma. Its absence in the normal outer cortex and theca interna, and in stromal hyperplasia and thecoma implies that sex hormones do not constitute a stimulus for alpha-SM actin production in the ovary. Among neoplasms it is most widely represented in the stroma of epithelial tumors in which it may reflect stromal stimulation mediated by neoplastic epithelium. PMID- 2567552 TI - Cytogenetic relationship between uterine lipoleiomyomas and typical leiomyomas. AB - The chromosomes from two human uterine lipoleiomyomas, L25 and L26, from the same patient, were studied by a banding technique applied to preparations from short term cultures. Both tumors displayed the same pseudodiploid stemline characterized by the reciprocal translocation t (5; 12) (q12; q24). These observations coincide with the previous finding that the largest subgroup of typical leiomyomas with an abnormal stemline are characterized by a long-arm change of one chromosome No. 12. The combined results support the previously advanced hypothesis that different histologic subtypes of uterine leiomyomas are derived from a common totipotential stem cell. This interpretation also fits with a proposed theory about the derivation of malignant leiomyomatous uterine neoplasms. PMID- 2567553 TI - [Hormones of the central nervous system--phylogenetic and integrated approach]. PMID- 2567554 TI - [Central regulation of adenohypophyseal function]. AB - The secretion of adenohypophyseal hormones is controlled by hypothalamic hypophysotropic hormones with stimulating (hormone releasing factors) or inhibitory (hormone release inhibiting factors) actions. The release of hypothalamic hormones is regulated by hierarchically higher nerve centres via neurons which liberate neurotransmitters at their endings. The secretion of growth hormone is controlled by hypothalamic hormones, somatotropin releasing factor and somatotropin release-inhibiting factor; of the neurotransmitters, the strongest effects have noradrenaline and dopamine. The release of ACTH is controlled by two stimulating hormones, the ACTH releasing factor and vasopressin, the effects of neurotransmitters are less marked, with the involvement of noradrenaline, serotonin, acetylcholine, gamma aminobutyric acid and other agents. Prolactin release is under the main inhibitory control of hypothalamic dopamine, no release-stimulating hypothalamic factor could be unequivocally demonstrated as yet; likely, several peptides are involved in this mechanism. The release of thyrotropic hormone is stimulated by thyrotropin releasing factor, whereas somatotropin release-inhibiting factor has an inhibitory action. Of the neurotransmitters, the inhibitory effect of dopamine is important; this agent however acts also at the hypophyseal level. External hypothalamic hormones and regulatory neurotransmitters are used in the diagnosis and treatment of neuroendocrine disorders. PMID- 2567555 TI - [Metoprolol and metipranolol in the treatment of hypertension-- comparison of the effects of a single dose and administration for 1 month]. AB - In six subjects the hypotensive action of metoprolol (Betaloc, Egis) after a dose of 166.7 +/- 47.1 mg and metipranolol (Trimepranol, Spofa) 21.7 +/- 3.7 mg was investigated. Betaloc influences after one month's administration the systolic blood pressure more effectively in a recumbent position and upright position and the diastolic pressure in a sitting position and after walking. The authors investigated also indicators of the circulation, blood sugar values, values of immunoreactive insulin, aldosterone and plasma renin activity after a single dose of 10 mg Trimepranol or 100 mg Betaloc. Betaloc caused a significant drop of the systolic blood pressure already after 15 minutes, Trimepranol after 30 minutes, in both instances the maximum effect was recorded after 45 minutes and persisted throughout the six-hour investigation. The diastolic blood pressure and heart rate were influenced only little. The blood sugar levels after Betaloc rose significantly during the first hour, after both drugs during the third hour. No correlation was found between changes of the blood sugar level and immunoreactive insulin levels. No differences were revealed as regards the effect on plasma renin activity and aldosterone levels. PMID- 2567556 TI - Possible neuroleptic-induced resistance to the human immunodeficiency virus. PMID- 2567557 TI - Management of marine stings and scrapes. PMID- 2567558 TI - [Quantifying mineralization processes in post-traumatic algodystrophy using computerized tomography densitometry]. AB - Computed tomography examinations have been made in 12 patients with posttraumatic algodystrophy. Significant differences of sides were found to be a measure of demineralisation by CT densitometry. 7 patients could be repeatedly examined during up to 60 weeks after the trauma. CT-integrated densitometry enables to quantify the process of de- and remineralisation and to objectify the disease according to the spongiosa bone. It seems to be possible to value the process of mineralisation by this method. PMID- 2567559 TI - Fatal intoxications in the Nordic countries. A forensic toxicological study with special reference to young drug addicts. AB - Fatal intoxications in the 15-34 age group in the five Nordic countries during the years 1984 and 1985 (Sweden only in 1984) were investigated. The known drug addicts were studied separately. The highest incidence of intoxications, calculated per 10(5) population, was found in Finland (11.3), followed by Denmark (10.3), Sweden (8.5), Iceland (7.2) and Norway (6.6). The percentage of intoxications caused by drugs was 92 in Denmark, 71 in Norway, 66 in Sweden, 50 in Finland and 17 in Iceland. Ethanol intoxications were seen 5-7 and 2-3 times as frequently in Finland and in Iceland, respectively, than in the other three countries. Carbon monoxide intoxications accounted for two-thirds of all fatal intoxications in Iceland. Drug addicts accounted for 62% of all fatal intoxications in the Danish material. The corresponding figures were 33% in the Norwegian, 16% in the Swedish and 5% in the Finnish material. No deaths in drug addicts were found in Iceland. Most drug addicts in Denmark, Norway and Sweden died of hard drugs and most in Norway and Sweden, from heroin or morphine, whereas in Denmark other strong analgesics, such as methadone, dextropropoxyphene and ketobemidone, accounted for 40% of all hard-drug-related fatal intoxications. To a certain extent the results reflect differences in the legal autopsy routines in the various Nordic countries. However, the ascertainment of drug addicts is assumed to be near-complete in each country. PMID- 2567560 TI - Clinical effects of azaperone-metomidate, as compared to propionylpromazine xylazine-metomidate or xylazine-ketamine combinations in anaesthesia of dogs. AB - Eighteen dogs of Tanzanian breeds divided into three groups of 6 were anaesthetized using either azaperone-metomidate (2 mg/kg, i.m. and 10 mg/kg i.p., respectively), propionyl promazine-xylazine-metomidate (2 mg/kg i.m., 1 mg/kg i.m. and 10 mg/kg i.p., respectively), or xylazine-ketamine (1 mg/kg i.m. and 11 mg/kg i.m., respectively). The clinical effects on respiration rate, heart rate and body temperature were studied until recovery. Hypersensitivity to noise was associated with azaperone metomidate anaesthesia. The other combination produced a smooth and uneventful induction and recovery from anaesthesia. Muscle relaxation and analgesia were adequate in all groups. Duration of xylazine ketamine anaesthesia was shortest (30 +/- 5 minutes) followed by azaperone metomidate (50 +/- 15 minutes) and the longest duration was with propionyl promazine-xylazine-ketamine (120 minutes). Azaperone and metomidate was associated with marked increases in cardiac and respiration rates and marked hypothermia, which persisted throughout. Minimal changes were observed in the other combinations. Azaperone-metomidate seems to be preferable due to the moderate period of anaesthesia adequate for most operations. However, all the three combinations offer a practical application because of the convenient route of administration. PMID- 2567561 TI - The evocation of rumination in sheep by the close-arterial injection of catecholamines, cholinergic drugs, autacoids and gastrointestinal hormones into the forestomach. AB - The effect of alpha-2 adrenoreceptor agonists, cholinergic drugs, autacoids and some gastrointestinal hormones upon the evocation of rumination in sheep when injected by close-arterial injection into the forestomach was examined. Apart from adrenaline, noradrenaline, dopamine and xylazine only one alpha-2 agonist (BHT933) evoked rumination effectively. Acetylcholine, neostigmine, the gastrointestinal hormones and the autacoids examined did not evoke rumination consistently. Tyramine did not usually evoke rumination and guanethidine injected before adrenaline did not prevent the latter from evoking rumination. The evocation of rumination by catecholamines does not appear to involve sympathetic, cholinergic or non-sympathetic non-cholinergic neurones in the enteric nervous system to activate the nervous sensory receptors involved in the reflex evocation of rumination. PMID- 2567562 TI - Involvement of D1 dopamine receptors in the control of TSH secretion in the male rat. AB - The effects of SCH 23390 (D1 dopamine receptor antagonist), SK&F 38393 (D1 dopamine receptor agonist), raclopride and remoxipride (D2 dopamine receptor antagonists) and ketanserin (5-hydroxytryptamine 2 receptor antagonist) on TSH serum levels (radioimmunoassay) and on brain catecholamine levels (Falck-Hillarp methodology in combination with quantitative histofluorimetry) were studied. SCH 23390 produced a dose-dependent increase in serum TSH levels in the lower dose range (0.01-0.03 mg kg-1, i.p.) administered 30 min before decapitation and in the higher dose range (1.0-3.0 mg kg-1) when given 2 h before decapitation. Following 30 min of treatment with the high doses of SCH 23390, reductions in serum TSH levels were found. The changes observed following SCH 23390 treatment occurred without affecting catecholamine levels in the median eminence and the peri- and paraventricular hypothalamic regions. Raclopride (0.1-10 mg kg-1, i.p.), remoxipride (1.0 mg kg-1, i.p.) or ketanserin (0.3 mg kg-1, i.p.) changed neither serum TSH levels nor brain catecholamine levels, SK&F 38393 (1.0-10 mg kg 1, i.p.) produced an increase in serum TSH levels. The results suggest the existence of inhibitory and facilitatory mechanisms regulating TSH secretion mediated via D1 dopamine receptors. PMID- 2567563 TI - Evidence for sympathetic regulation of the hydraulic conductance of rat jejunal mucosa in vivo. AB - The aim of the study was to test the hypothesis that sympathetic nerve stimulation inhibits jejunal fluid secretion by decreasing the hydraulic conductance (Lp) of the mucosa. Jejunal segments of anaesthetized rats were mounted in a specially constructed chamber which makes it possible to measure net fluid transport together with transmural potential difference or short circuit current in vivo. Lp was estimated by measuring the effects on net fluid transport rate elicited by stepwise changes in luminal pressure. Lp was measured in four groups of animals: (1) under control conditions; (2) after hexamethonium administration, to block intramural secretory neurons; (3) during efferent stimulation of the mesenteric nerves, to activate sympathetic neurons, and (4) during mesenteric nerve stimulation (MNS) after pre-treatment with phentolamine, an alpha-adrenergic antagonist, to block the sympathetic effector mechanism. Lp measured at positive luminal pressures in control animals was approximately 4 microliters min-1 cmH2O-1 100 cm-2 serosal surface area. When measured at negative luminal pressures, this value increased approximately threefold. Hexamethonium did not significantly influence Lp. At positive luminal pressures, MNS decreased Lp to a value not significantly different from zero. This effect was abolished by phentolamine. At negative luminal pressures, MNS decreased Lp by approximately 50% and this effect was partly antagonized by phentolamine. The results suggest that a decrease in mucosal hydraulic conductance may contribute to the antisecretory effect of sympathetic nerve stimulation in vivo. PMID- 2567564 TI - Evidence for an intramural nervous control of epithelial cell migration in the small intestine of the rat. AB - The migration of epithelial cells along the crypt-villus axis in the small intestine of the rat was followed by labelling epithelial cells during mitosis with [3H]thymidine given i.v. Using two different techniques (autoradiography and determination of tissue radioactivity) it was demonstrated that 6-9 h after giving the tracer the labelled cells had migrated longer in intestinal segments exposed to cholera toxin than in control segments. This effect of cholera toxin was abolished by giving hexamethonium. We have earlier shown that cholera toxin induces fluid secretion to a large extent by activating the enteric nervous system and we conclude from the present observations that cholera toxin in a similar fashion exerts a trophic effect on the intestinal epithelium via intramural nervous reflexes. The importance of co-release of several neurotransmitters in explaining the trophic effect is tentatively discussed. PMID- 2567565 TI - In-vivo quantification of myocardial Na-K pump rate during beta-adrenergic stimulation of intact pig hearts. AB - Maintenance of adequate electrical activity of the heart depends critically on the ability of the Na-K pump to compensate for normal passive sodium and potassium fluxes. Using sudden injections of [3H]ouabain into the left coronary artery in anaesthetized open-chest pigs, we monitored transient changes in myocardial potassium balance by PVC-valinomycin mini-electrodes. When related to the number of pumps blocked and fractional inhibition, these data provided estimates of total Na-K pump capacity as well as actual pump rate and perturbations of the Na-K balance. Experiments were performed in hearts with and without intracoronary isoprenaline infusion (2.5 nmol min-1). After injection of 120 nmol [3H]ouabain into the left coronary artery, myocardial [3H]ouabain concentrations were 118 (74-178) and 103 (76-145) pmol g-1 and total concentrations of [3H]ouabain binding sites were 893 (752-1076) and 785 (691-877) pmol g-1 (median, 95% confidence interval) in isoprenaline-treated and control hearts respectively (differences not significant). The [3H]ouabain injection caused a net potassium release of 81 (56-132) and 43 (23-75) mumol 100 g-1 (median, 95% confidence interval) in isoprenaline-treated and control hearts respectively (n = 6-8; significance of difference, P = 0.03). Na-K pump rate estimated from mono-exponential release curves was 6363 (3942-10,858) K+ ions min 1 site-1 during beta-adrenoceptor stimulation and 2514 (1380-4322) in control (significance of difference, P = 0.03). This corresponds to 40 and 16%, respectively, of the maximum possible pump rate determined from ATP hydrolysis. Comparison of accumulated potassium release and relative Na-K pump rate indicates that catecholamines enhance the sensitivity of the Na-K pump for intracellular sodium. PMID- 2567566 TI - Differential effect of bombesin on intraluminal and intravascular release of gastric gastrin and somatostatin in anaesthetized rats. AB - The aim of the present investigation was to study how bombesin, gastrin-17, cholecystokinin-8 (CCK-8) and electrical vagal stimulation influence the release of gastrin and somatostatin into the gastric lumen. Bombesin (3 and 30 nmol kg-1 h-1), gastrin-17 and CCK-8 (10 nmol kg-1h-1) were infused i.v. and vagal stimulations at 5 V, 2 ms, 5 Hz were performed in anaesthetized rats, in which the stomach was perfused with a dextran solution (pH approximately 6 or approximately 1.5). pH, gastrin and somatostatin levels were measured in the perfusate after having passed the stomach. In addition, blood samples were drawn from the jugular vein in the experiments in which bombesin was infused. Gastrin and somatostatin levels were determined with radioimmunoassay, and gastrin- and somatostatin-like immunoreactivity will be referred to as gastrin and somatostatin below. Infusion of bombesin, 3 and 30 nmol kg-1 h-1, did not influence acid secretion as evidenced by an unchanged intraluminal pH. Nor was the intraluminal secretion of gastrin or somatostatin influenced by bombesin, whereas plasma gastrin and somatostatin levels were significantly increased at the higher dose. In contrast, infusion of CCK-8 and gastrin-17 (10 nmol kg-1 h 1), as well as electrical vagal stimulation, significantly decreased the pH and increased the somatostatin levels in the perfusate. Vagal activation in addition increased the gastrin levels. The present results, demonstrating that bombesin influences plasma and perfusate levels of gastrin and somatostatin differently, indicate that intraluminal and intravascular gastrin and somatostatin release may be separately regulated. PMID- 2567568 TI - Edema and cell infiltration in the phorbol ester-treated mouse ear are temporally separate and can be differentially modulated by pharmacologic agents. AB - The temporal patterns of edema and accumulation of the PMN marker enzyme, myeloperoxidase (MPO), were examined following application of tetradecanoylphorbol acetate (TPA) to mouse ears. After application of 2.5 micrograms TPA, edema peaked at 6 hr, while MPO activity peaked at 24 hr. Pharmacological agents with defined mechanisms of action, delivered orally or topically, were assessed for effects on these responses. For oral administration, compounds were delivered 1 hr before and 6 hr after TPA and for topical administration compounds were delivered at 15 min and 6 hr after TPA. Topical and oral corticosteroids inhibited both edema and MPO accumulation. Cyclooxygenase and lipoxygenase inhibitors were very effective against MPO accumulation but were either inactive or moderately active vs edema. Anti-histamine/anti-serotonin agents had little effect on edema, but could inhibit or exacerbate MPO accumulation depending on dose and route of administration. Topically applied histamine itself did not effect TPA-induced edema, but markedly suppressed MPO accumulation. Acetone, the vehicle, when topically applied between 0.5 and 2 hr after TPA inhibited MPO accumulation by 60-80%, but had little effect on edema. Acetone applied before 0.5 hr or after 2 hr had no effect on either parameter. These results indicate that in the TPA-induced ear inflammation model the MPO response at 24 hr may be a useful additional indicator of drug activity. PMID- 2567569 TI - [A case of ureteral and vesical malacoplakia]. AB - A case of ureteral and vesical malacoplakia is reported. A 58-year-old woman was admitted for non-papillary sessile bladder tumor. Ultrasound examination revealed bilateral hydronephrosis and renal function was deteriorated. Escherichia coli and beta-streptococcus were isolated from her urine. Antegrade pyelography demonstrated obstruction of the bilateral ureters. Percutaneous pyeloscopy revealed a bean-sized yellow nodule at the upper portion of the right ureter. Pathohistological the lesions of aggregates of macrophage in which typical Michaelis-Gutmann bodies were found at the punch biopsies of both vesical and ureteral lesions. She was treated successfully with the administration of bethanechol chloride and antibiotics. This case seems to be the second report of ureteral malacoplakia in the Japanese literature. PMID- 2567567 TI - H1-antihistamines and calmodulin antagonists inhibit the ionophore A23187-induced eicosanoid formation by human leukocytes. AB - The effects of the H1-antihistamines astemizole, oxatomide and pyrilamine, of the calmodulin antagonists trifluoperazine and N-(6-aminohexyl)-5-chloro-1 naphthalenesulfonamide (W-7), on the ionophore A23187 (5 mumol/l)-induced release of cysteinyl-leukotrienes (LT) and thromboxane (TX) B2 from mixed human leukocytes were investigated in comparison to those of the cyclooxygenase inhibitor indomethacin and the lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA). In contrast to pyrilamine both astemizole and oxatomide inhibited the release of cysteinyl-LT and TXB2 with IC50 values between 4 and 23 mumol/l. Both astemizole and oxatomide were about twice as effective in inhibiting cysteinyl-LT release as compared to TXB2 release. Similar to astemizole and oxatomide the calmodulin antagonists trifluoperazine and W-7 inhibited the eicosanoid release. W-7 was, however, clearly less effective and in contrast to trifluoperazine no difference was observed in its potency to inhibit cysteinyl-LT or TXB2 release. The H1-antihistamines, astemizole and oxatomide as well as the calmodulin antagonists did not cause intracellular retention of the eicosanoids tested. The reference compounds indomethacin and NDGA proved to be the most potent inhibitors. The results demonstrate that the therapeutic antihistamines astemizole and oxatomide as well as the classical calmodulin antagonists trifluoperazine and W-7 are able to inhibit eicosanoid formation. PMID- 2567570 TI - [Study on urinary splitting enzymes and proteins. III. Effect of non-ionic contrast medium on renal function]. AB - Renal toxicity of non-ionic contrast medium (iohexol) for drip infused pyelography (DIP) was studied in a randomized trial of nine patients with normal renal function. Urine samples were collected before and immediately after DIP, and analyzed for albumin, an index of glomerular permeability; gamma-glutamyl transpeptidase (gamma-GTP), a brush-border enzyme; N-acetyl-beta-glucosaminidase (NAG), a lysosomal enzyme; alpha 1 microglobulin (alpha 1MG) and beta 2 microglobulin (beta 2MG), an index to tubular proteinuria; and creatinine. The urinary excretion of enzymes and proteins was compared with urinary creatinine. Urinary excretion of gamma-GTP and NAG increased significantly (P less than 0.001, 0.02) after DIP. Urinary alpha 1 MG and beta 2-MG did not change significantly. The change of urinary albumin was mild. Our data suggest that non ionic, low osmolal radiocontrast medium ioheol shows a lower renal tubular toxicity, and the brush-border enzyme gamma-GTP and lysosomal enzyme NAG are considered as a good index for renal tubular damage. PMID- 2567571 TI - Sucralfate maintenance therapy in duodenal ulcer disease. A review. AB - There are a large number of patients with chronic duodenal ulcer disease who warrant long-term maintenance therapy to diminish the risk of recurrence and thereby the risk of further complications such as gastrointestinal bleeding. The efficacy of sucralfate has been compared with both placebo and histamine (H2) receptor antagonists and sucralfate in a dose of 1 g twice a day or 2 g taken at night. It is a safe and effective medication in preventing duodenal ulcer recurrence. However, duodenal ulcer relapse rates always exceed 20 percent and frequently approach 50 percent, whether the therapy be H2-receptor antagonists or sucralfate, and the use of dosages that are half the healing dose seems irrational. It would therefore seem reasonable to continue maintenance therapy at the healing dose, whatever medication is used. Any increased costs for drugs should be outweighed by savings in indirect costs. PMID- 2567572 TI - Side effects of stress bleeding prophylaxis. AB - Conventional stress bleeding prophylaxis with antacids or histamine (H2) antagonists, as well as the newer mucosa-protective drugs pirenzepine and sucralfate, are satisfying most of the clinicians with regard to efficacy of stress bleeding prevention. Therefore, potential side effects are attaining crucial importance with regard to the drugs to be used. Pharmacologic blockade of cardiac H2-receptors increases the risk of bradycardia and negative inotropic effects as well as coronary vasoconstriction at least in the presence of elevated plasma histamine levels. Intracardiac injection of pirenzepine can lead to temporary tachycardia. Elderly patients have been shown to be at an increased risk of side effects to the central nervous system when treated with H2 antagonists. These drugs can also induce toxic effects in the liver. Cimetidine leads to interactions with a number of drugs used in the intensive care unit. In patients with pre-existing pulmonary diseases, H2-antagonists have been demonstrated to increase pulmonary bronchoconstriction. Alkalinization of the gastric juice is associated with a significant increase in colonization of gram negative bacteria in the stomach. In intubated patients, aspiration of stomach contents occurs in 30 to 40 percent of the patients. A number of studies have shown a direct correlation between alkalinization of the gastric juice and pulmonary infections. Sucralfate and to a lesser degree pirenzepine can reduce the risk of pulmonary infections. Sucralfate also exerts a bactericidal effect. Recent investigations support the hypothesis that alkalinization of the stomach also increases the risk of systemic infections. This may be the main reason for the observation that at least in ventilated patients sucralfate, unlike H2 antagonists or antacids, leads to a significant reduction of the mortality rate compared with conventional stress bleeding prophylaxis. PMID- 2567574 TI - Preeclampsia: a hyperdynamic disease model. AB - Preeclampsia is a common disease and as such is a significant contributor to maternal and neonatal morbidity and mortality. Despite the ubiquity of the disease and its public health impact, no comprehensive mechanism has been established. Therapy has been limited to bed rest and premature delivery. The purpose of this article is to examine preeclampsia from a hemodynamic point of view and to advance a hypothesis of disease mechanism. PMID- 2567573 TI - A national survey on preterm labor. AB - A national survey was conducted to examine the American obstetricians' approach to management of preterm labor. Multiple-choice questionnaires were mailed to all current members of the Society of Perinatal Obstetricians and 750 randomly selected diplomates of the American College of Obstetricians and Gynecologists. Respondents included 249 members of the Society of Perinatal Obstetricians and 441 fellows of the American College of Obstetricians and Gynecologists. Analysis of the replies reflects wide variations among American obstetricians' criteria to diagnose preterm labor, choice of tocolytic agent, use of amniocentesis, and use of corticosteroids. Significant differences were found between the two groups in many aspects of diagnosis and management of preterm labor. However, the replies reflect a high degree of awareness of and many instances of first hand experience with severe beta-adrenergic therapy complications, including maternal death. These findings emphasize the need for caution in selection of patients for beta adrenergic tocolytic therapy and judicious use of these agents. PMID- 2567575 TI - Impairment of glucose disposal by infusion of triglycerides in humans: role of glycemia. AB - The present study was designed to assess the role of hyperglycemia (150 mg/dl) vs. euglycemia (90 mg/dl) on glucose metabolism in vivo during the infusion of a triglyceride emulsion (Intralipid). Seven young healthy volunteers were studied on four occasions using the hyperinsulinemic clamp technique, twice during euglycemia and twice during hyperglycemia, without or with Intralipid. Glucose oxidation (O) was calculated from continuous respiratory exchange measurements, and glucose storage (S) was obtained as the difference between total glucose disposal (M) and O. Two-way analysis of variance with interaction term demonstrated 1) a significant increase for M with hyperglycemia and a decrease with Intralipid; no interaction, and 2) in euglycemia, O/M and S/M occurred in one-to-one ratios; on the other hand, during 150-mg/dl hyperglycemia, the ratio dropped roughly to 1:2. Intralipid had no effect on the ratio, and no interaction could be observed. These results suggest the existence of physiological regulatory mechanisms by which 1) the rise in plasma free fatty acid inhibits both oxidative and nonoxidative glucose disposal, and 2) the rise in glycemia stimulates predominantly nonoxidative glucose disposal. PMID- 2567577 TI - Cardiovascular effects of dynorphin A-(1-13) and arginine vasopressin in fetal lambs. AB - The cardiovascular effects of the kappa-opioid receptor agonist, dynorphin A-(1 13) (D13), were compared with those of arginine vasopressin (AVP) in intact and bilaterally vagotomized (VGX) fetal lambs. Intravenous injection of AVP (114 ng/kg) produced a significant rise in mean arterial pressure (MAP) lasting 15-20 min in both intact and VGX lambs. AVP produced a bradycardia in intact fetuses concurrently with the MAP response and had no effect on heart rate (HR) in VGX fetuses. D13 (500 micrograms/kg) also produced significant increases in MAP in intact and VGX fetuses that lasted 45-60 min. D13 produced a bradycardia in intact fetal lambs but, unlike AVP, significantly increased HR in VGX fetuses. HR responses to D13 had time courses similar to MAP responses in both intact and VGX fetal lambs. Pretreatment with a vasopressin V1-receptor antagonist significantly attenuated pressor responses to AVP in both treatment groups and to D13 in intact fetuses and abolished the D13 pressor response in VGX fetuses. The antagonist also completely blocked HR responses to AVP in intact and to D13 in VGX fetuses and attenuated the D13 HR response in intact fetuses. Therefore the effects of D13 on ovine fetal cardiovascular function appear to be mediated in part through AVP pathways. D13 also appears to have a positive chronotropic effect on the heart that is normally masked by inhibitory vagal activity. PMID- 2567576 TI - Demonstration of a role for growth hormone in glucose counterregulation. AB - To test the hypothesis that growth hormone secretion plays a counterregulatory role in prolonged hypoglycemia in humans, four studies were performed in nine normal subjects. Insulin (15 mU.M-2.min-1) was infused subcutaneously (plasma insulin 27 +/- 2 microU/ml), and plasma glucose decreased from 88 +/- 2 to 53 +/- 1 mg/dl for 12 h. In study 1, plasma glucose, glucose fluxes (D-[3-3H]glucose), substrate, and counterregulatory hormone concentrations were simply monitored. In study 2 (pituitary-adrenal-pancreatic clamp), insulin and counterregulatory hormone secretions (except for catecholamines) were prevented by somatostatin (0.5 mg/h iv) and metyrapone (0.5 g/4 h po), and glucagon, cortisol, and growth hormone were reinfused to reproduce the concentrations of study 1. In study 3 (lack of growth hormone increase), the pituitary-adrenal-pancreatic clamp was performed with maintenance of plasma growth hormone at basal levels, and glucose was infused whenever needed to reproduce plasma glucose concentration of study 2. Study 4 was identical to study 3, but exogenous glucose was not infused. Isolated lack of a growth hormone response caused a decrease in hepatic glucose production and an increase in glucose utilization that resulted in an approximately 25% greater hypoglycemia despite compensatory increases in plasma catecholamines. Plasma free fatty acid, 3-beta-hydroxybutyrate, and glycerol concentrations were reduced approximately 50%. It is concluded that growth hormone normally plays an important counterregulatory role during hypoglycemia by augmenting glucose production, decreasing glucose utilization, and accelerating lipolysis. PMID- 2567578 TI - Increase in sympathetic outflow by paraventricular nucleus stimulation in awake rats. AB - Our previous studies demonstrated that stimulation of the hypothalamic paraventricular nucleus (PVN) in anesthetized rats evoked a depressor response accompanied with a decrease in sympathetic outflow (H. Kannan, A. Niijima, and H. Yamashita, J. Auton. Nerv. Syst. 19: 83-86, 1987; H. Yamashita, H. Kannan, M. Kasai, and T. Osaka, J. Auton. Nerv. Syst. 19: 229-234, 1987). Because anesthesia may alter cardiovascular responses, we examined in conscious rats the effects of PVN stimulation on arterial pressure, heart rate, and renal sympathetic nerve activity. Electrical stimulation through chronically implanted electrodes evoked increases in arterial pressure and renal sympathetic nerve activity with a slight decrease in heart rate. The magnitude of responses was dependent on the frequency and the intensity of the stimulus. Latency of the excitatory response of the renal sympathetic nerve activity was approximately 70 ms. Microinjection of L glutamate (0.5 M, 200 nl) into the PVN area also elicited increases in blood pressure and renal sympathetic nerve activity. These results suggest that activation of PVN neurons in conscious rats produces pressor responses due to an increase in the sympathetic outflow. These findings contrast with those obtained previously in anesthetized rats. PMID- 2567580 TI - [Interactions of H2 antagonists and non-depolarizing muscle relaxants]. AB - Many drugs potentiate the action of non depolarizing relaxants. These interactions are of clinical importance if such drugs are administered during the perioperative period. H2 Antagonists are increasingly often used for premedication. Cimetidine inhibits the elimination of a number of drugs used in the perioperative period. We therefore investigated whether H2 antagonists enhanced neuromuscular blockade by vecuronium, a medium short acting non depolarizing muscle relaxant. METHODS. The study was carried out in 24 female patients (ASA class I or II) scheduled for microsurgical procedures. Neuromuscular transmission was recorded electromyographically using four stimulations every 20 s to the ulnar nerve. After induction with thiopentone, anesthesia was maintained with fixed concentrations of volatile anesthetics. Fentanyl was administered for additional analgesia. Vecuronium was used as the sole muscle relaxant. Fixed repetitive doses of vecuronium (0.8-1.2 mg) were injected whenever the T1 returned to 25%. This time interval was defined as the T1-25 period. The study proper started when the T1-25 period had stabilized. After two control periods, six patients in each group received either 200 or 400 mg cimetidine or 100 mg ranitidine. The fourth group was the control group. The T1-25 periods and the maximal EMG depression were recorded automatically for at least two further periods. The first measured period was recorded as 100% and the length of each other periods was calculated as a percentage of the control period. This method enables an intraindividual comparison of the length of the T1 25 period and the maximal EMG depression before and after administration of the H2 antagonists. A two-tailed Student's t-test was used to test statistical significance, P less than 0.05 being accepted as significant. RESULTS. In the control group and in the group with 200 mg cimetidine or 100 mg ranitidine no statistical significant prolongation of the T1-25 period or of the maximal EMG depression could be observed, while after 400 mg cimetidine there was significant prolongation (mean 161 +/- 14.8%) of the T1-25 period and significantly greater EMG depression compared with the pre-cimetidine values. In the groups with 200 mg cimetidine or 100 mg ranitidine few patients showed prolongation of the T1-25 period up to 130%. DISCUSSION. Our results confirm experimental studies that have shown cimetidine to enhance aminoglycoside--relaxant interactions. Because we found an immediate response to the administration of the H2 antagonists, the interaction cannot be on the elimination side; it must be at the neuromuscular junction. Experimental investigation has shown that calcium reverses the cimetidine effects. It is therefore probable that the cimetidine--relaxant interaction occurs at the presynaptic level. Careful observation seems to be necessary if H2 antagonists, especially cimetidine, are administered intraoperatively at the same time as drugs that also enhance PMID- 2567579 TI - Effects of topical treatment with H1 and H2 antagonists on clinical symptoms and nasal vascular reactions in patients with allergic rhinitis. AB - Fifteen asymptomatic subjects with allergic rhinitis participated in a double blind, randomized, crossover, placebo-controlled study. The subjects were pretreated intranasally with a single dose of a selective H1 receptor antagonist, levocabastine, and/or selective H2 receptor antagonist, ranitidine, prior to a nasal allergen challenge. The nasal symptoms obtained at the challenge were assessed using a scoring technique 15 min after the allergen exposure. The nasal airway resistance was determined twice prior to and once after the allergen challenge using anterior rhinomanometry. The nasal mucosal blood flow was determined before and 15 min after allergen challenge using the 133Xe wash-out technique. After pretreatment with the H1 antagonist there was a statistically significant reduction in the number of sneezes and rhinorrhea compared to pretreatment with placebo. Pretreatment with the H2 receptor significantly decreased the rhinorrhea but not the sneeze. The nasal blockage was unaffected by both the H1 and the H2 antagonists. Pretreatment with the H1 and/or the H2 antagonists inhibited the reduction in the nasal mucosal blood flow induced by the allergen challenge to a significant degree. The present findings suggest that topical treatment with the highly selective histamine antagonist, levocabastine, inhibits allergen-induced reflex-mediated symptoms. H1 and H2 receptors do not appear to be involved in the regulation of the tone of the capacitance vessels. This indicates that a more complex mechanism participates in the induction of nasal blockage than the direct effect of histamine on H1 and H2 receptors on the capacitance vessels of the nasal mucosa alone. Both H1 and H2 receptors are of importance for the regulation of nasal mucosal blood flow during the allergic reaction. PMID- 2567581 TI - [Vecuronium in dystrophia myotonica (Curschmann-Steinert)]. AB - An emergency laparotomy was performed in a 31-year-old female (body wt 48 kg) with known myotonic dystrophy. Premedication with dantrolene (1 mg/kg i.v.) was used to prevent a myotonic response. Muscle relaxation was monitored electromyographically. Following induction with fentanyl (0.3 mg) and thiopental (200 mg), muscle relaxation was achieved with 2 mg vecuronium titrated for about 3 min until the T1-response was reduced to 10%. The recovery time was normal. A repetitive dose of 0.5 mg vecuronium was necessary after 20 min, when the T1 reached 60%. Extubation and the early postoperative period were uneventful. Because of the unknown predisposition of our patient for the development of malignant hyperthermia, anesthesia was performed with trigger-free anesthetics. PMID- 2567583 TI - Does nitrous oxide affect the hemodynamic effects of anesthesia induction with propofol? AB - Anesthesia was induced in 20 patients, ASA physical status I and II, with either propofol (2.5 mg.kg-1), vecuronium (100 micrograms.kg-1), and 100% oxygen (Group A), or with equal doses of propofol and vecuronium but with 70% nitrous oxide in oxygen (Group B). All patients were premedicated with lorazepam 2 mg orally. In both groups systolic arterial pressure decreased after 3 minutes (P less than 0.05) due to decreases in cardiac output and stroke volume (P less than 0.05). Systemic vascular resistance in both groups did not change immediately after administration of propofol but increased (P less than 0.05) following intubation. Addition of nitrous oxide did not alter hemodynamic parameters associated with propofol induction. PMID- 2567584 TI - Tetanic fade following administration of nondepolarizing neuromuscular blocking drugs. AB - Fade in response to tetanic stimulation was studied following administration of atracurium 120 or 225 micrograms/kg, vecuronium 23 or 40 micrograms/kg, pancuronium 30 or 60 micrograms/kg, or d-tubocurarine 185 or 450 micrograms/kg. Ten patients received each dose and tetanic fade was measured at maximum block in the patients, who received the lower doses of the relaxants or at 10% recovery in those who received the higher doses. Fade during tetanic stimulation was generally similar in all the groups with the exception of the higher dose of pancuronium which showed a significantly greater fade in comparison with the higher doses of atracurium and d-tubocurarine. If fade in response to tetanic stimulation represents a prejunctional effect, the results from the present study suggest that neuromuscular blocking drugs cannot be differentiated with respect to their relative prejunctional effects by measurement of tetanic fade during established block after administration of clinically useful doses as used in the present study. PMID- 2567582 TI - NCAM and Thy-1 in special sense organs of the developing mouse. AB - The distribution of the neural cell adhesion molecule (NCAM) and Thy-1 in the olfactory mucosa and olfactory bulb, the eye and the inner ear was examined with immunocytochemistry in mouse embryos from embryonic day 12 (E 12) to embryonic day 19 (E 19). In general, neurons are completely outlined with NCAM, whereas Thy 1 outlines only dendrites and axons. A variable cytoplasmic staining for Thy-1 is present in the perikarya. Neurons directly associated with special sense organs express NCAM and Thy-1 already from the earliest stage and throughout the period investigated, apart from the olfactory neurons in which Thy-1 disappears at E 19. The mitral cells in the olfactory bulb show Thy-1 but no NCAM reactivity. In the eye, lens fibers express Thy-1 and the pigmented layer expresses NCAM; neither of the two molecules can be detected at E 19. In the inner ear, hair cells express NCAM at E 19. Based on the distribution during the developmental period studied and on the cellular localisation of reaction products, it is suggested that the NCAM adhesion function could be of a more general nature by keeping appropriate cell membranes in close contact and thereby allowing more specific molecular interactions to take place. Thy-1, which is located on dendrites and axons, could be such a specific factor and function as recognition molecule in the developing nervous system. PMID- 2567585 TI - Drugs and the immune response: knowledge and perspectives in allergology. PMID- 2567586 TI - [Consensus development conference in resuscitation and emergency medicine: prevention of stress gastroduodenal hemorrhage]. PMID- 2567587 TI - [Consensus development conference in resuscitation and emergency medicine: management of severe acute asthma in adults]. PMID- 2567588 TI - 69 nations to attend ICN congress. PMID- 2567589 TI - Famotidine therapy for active duodenal ulcers. A multivariate analysis of factors affecting early healing. AB - OBJECTIVE: To identify factors that influence the rate of healing of duodenal ulcers. DESIGN: A stepwise multivariable statistical analysis of patients with duodenal ulcer in a multicenter, prospective, open-label study. Healing was assessed by endoscopy at 4 or 8 weeks. Antacid use and symptoms were recorded in a daily diary. SUBJECTS: Of 135 patients, ages 19 to 86, 50% had a previous duodenal ulcer, 46.7% smoked, and 34.8% had melena or hematemesis. INTERVENTIONS: Famotidine, 40 mg orally, at bedtime for 4 or 8 weeks depending on endoscopic evaluation of ulcer healing. Limited antacid use was permitted. SETTING: Office practices, hospital practices, and university-based medical centers. MEASUREMENTS AND MAIN RESULTS: Multivariable analysis identified five independent predictors present at the time of diagnosis that influenced ulcer healing. The odds of not healing for each risk factor after simultaneous adjustment of the other risk factors were as follows: alcohol use, 6.5 (CI, 2.0 to 20.7, P less than 0.002); ulcer size greater than 10 mm, 4.2 (CI, 1.5 to 11.6, P less than 0.005), bleeding symptoms. 3.5 (CI, 1.2 to 10.2, P less than 0.03); and a previous duodenal ulcer, 3.1 (CI, 1.05 to 9.0, P less than 0.04). The use of salicylates or nonsteroidal anti-inflammatory drugs before treatment was associated with an improved odds of healing (adjusted odds ratio, 0.2; CI, 0.1 to 0.9, P less than 0.04). The percentage of patients achieving complete ulcer healing after 4 weeks of famotidine decreased inversely with the number of risk factors present, ulcer size, and the quantity of daily alcohol use (P less than 0.001). Fewer than half of those patients who still had severe pain at day 7 achieved healing at 4 weeks (P less than 0.001). In contrast, smoking and 23 other factors had no statistically discernible effect on ulcer healing with famotidine. CONCLUSIONS: Five variables present at the time of diagnosis independently influenced the rate of ulcer healing at 4 weeks: alcohol use, ulcer size, bleeding symptoms, a previous duodenal ulcer, and previous use of salicylates or nonsteroidal anti inflammatory drugs. PMID- 2567590 TI - [Myocardial infarction: secondary prevention with betablockers]. AB - Invasive or pharmacological treatments at the onset of a myocardial infarction or a few days later, make it possible to decrease the mortality and morbidity through prevention of the ventricular fibrillation, decrease of the size of the infarction, inhibition of platelet aggregation and other mechanisms. Early administration of betablockers results in a significant decrease of the mortality at one year, in patients undergoing an active treatment. Later treatment also results in decreased coronary mortality and morbidity. This article intends to review the most recent information concerning the potential preventive value of beta-blockers after myocardial infarction. PMID- 2567592 TI - Porcine D-amino acid oxidase: production of the biologically active enzyme in Escherichia coli. AB - DNA molecules coding either for mature porcine D-amino acid oxidase or for truncated forms of the enzyme have been obtained by stepwise addition of synthetic oligonucleotides to a partial cDNA. Under the control of the lambda PL thermoregulatable promoter, these DNAs were respectively expressed in Escherichia coli as 36, 28 and 25 kilodalton polypeptides, specifically recognised by antibodies raised against the natural enzyme. None of the truncated proteins were biologically active whereas the mature recombinant species was able to hydrolyze D-alanine in vitro as efficiently as the natural product. PMID- 2567591 TI - Duration of a clozapine trial in neuroleptic-resistant schizophrenia. PMID- 2567593 TI - Gene expression of PCNA/cyclin in adult tissues and the R3230AC mammary tumor of rat. AB - We have investigated the gene expression of PCNA (Proliferating Cell Nuclear Antigen)/cyclin in rat tissues and the R3230AC mammary tumor. The steady-state mRNA level of PCNA/cyclin in a tissue is related to the proliferation of the tissue. The observation was confirmed with the results from the studies of the immunoblotting analyses and the DNA polymerase activity measurements. Furthermore, an overexpression of PCNA/cyclin was found in the R3230AC mammary tumor, which is accompanied by an altered PCNA/cyclin gene structure detected with the Southern blot analysis. PMID- 2567594 TI - Biosynthesis of nitric oxide from L-arginine. A pathway for the regulation of cell function and communication. PMID- 2567595 TI - Blockade by lipoxygenase inhibitors of Ca2+-dependent insulin secretion from permeabilized rat islets. A molecular mechanism distinct from that of alpha 2 adrenergic agonists. AB - To evaluate the regulation and effects of pancreatic islet lipoxygenase, adult rat islets were permeabilized, using digitonin or staphylococcal alpha-toxin, and then were studied in a medium simulating an intracellular milieu at fixed ambient concentrations of Ca2+. Permeabilized islets retained 12-lipoxygenase activity, as indicated by conversion of tritiated arachidonic acid to a predominant peak of [3H]12-hydroxyeicosatetraenoic acid (12-HETE); this activity was inhibited (89 98%) by the lipoxygenase blockers nordihydroguaiaretic acid (35 microM), BW755c (250 microM) or ETYA (35 microM). Lesser amounts of compounds coeluting with 15- and 11-HETE (but little or no 5-HETE) were formed; however, 11-HETE (and possibly some 15-HETE) was probably synthesized (at least in part) via cyclooxygenase, as suggested by the partial synthesis blockade induced by 50 microM ibuprofen. The production of 12-HETE did not require the presence of Ca2+, Mg2+ or ATP; it also was not stimulated by addition of cyclic AMP, a phorbol ester, or calmodulin. However, it was augmented modestly by provision of a basal cytosolic free Ca2+ concentration of 60-80 nM, with no further increase at physiologically elevated levels of 260-530 nM. Elevations in cytosolic free Ca2+ concentrations induced insulin release which was inhibited by cooling, epinephrine or protein kinase inhibitors and, therefore, was exocytotic in nature. Lipoxygenase inhibitors blocked this insulinotropic effect of calcium at submaximal or saturating Ca2+ concentrations (with or without its potentiation by 12-O-tetradecanoylphorbol-13 acetate, an activator of protein kinase C) by 53-82%. However, they did not reduce the Ca2+-independent secretory effects (at subnanomolar Ca2+ concentrations) of the phorbol ester alone. Similar results were seen using dibutyryl cyclic AMP to activate protein kinase A. The alpha 2-adrenergic agonists epinephrine or clonidine inhibited Ca2+-, TPA- or cyclic AMP-induced insulin release without reducing HETE formation. We conclude that (1) islet lipoxygenase is constitutively expressed and is not physiologically regulated by alpha 2-adrenergic agonism, Ca2+ or protein kinases; (2) lipoxygenase modulates insulin release; HETE production is not merely an epiphenomenon reflecting the activation (or inhibition) of exocytotic secretion; (3) islet lipoxygenase inhibitors reduce insulin secretion, at least in part, by blocking the direct effects of Ca2+ on exocytosis and/or its synergism with Ca2+-binding proteins such as protein kinase C; and (4) these same inhibitors do not directly poison protein kinase C or A, or the exocytotic apparatus.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2567596 TI - High affinity, saturable [3H]mepyramine binding sites on rat liver plasma membrane do not represent histamine H1-receptors. A warning. AB - Rat liver plasma membrane contains a saturable, high affinity binding site for the labelled histamine H1-antagonist [3H]mepyramine. Nonlinear regression analysis of the performed saturation experiments revealed an equilibrium dissociation constant (Kd) of 7.7 +/- 0.4 nM and a maximal binding capacity (Bmax) of 70.4 +/- 9.5 pmol/mg protein. Specific binding could be inhibited completely by several histaminergic ligands. However, the affinities of the tested H1-antagonists other than mepyramine for this binding site were quite low and the known stereospecificity displayed by the histamine H1-receptor for the enantiomers of chlorpheniramine and pheniramine was not found. Moreover, the H2 selective agonist 4-methylhistamine (Ki = 412 microM) was even more potent than its H1-selective 2-methylderivative (Ki = 772 microM). Since several ethylenediamines were also very potent in displacing [3H]mepyramine we suggest the presence of an ethylenediamine recognition site on rat liver plasma membrane which is unrelated to the histamine H1-receptor. It is stressed that a proper pharmacological characterization of a reported binding site is needed, since we show in this study that [3H]mepyramine, which is frequently used in studies concerning the H1-receptor, labels non-H1-receptor binding sites in rat liver plasma membrane. PMID- 2567597 TI - Effects of a beta-adrenergic agonist on protein turnover in muscle cells in culture. AB - beta-Adrenergic agents powerfully stimulate muscle growth in animals. Whether the mechanism of action involves a direct effect on muscle cell beta-receptors or is secondarily due to a beta-induced alteration in the hormonal environment is not known. To assess whether direct beta-receptor activation results in muscle protein accretion, we examined the effect of the beta-agonist zinterol on several anabolic processes in L8 muscle cells in culture. In vivo feeding of zinterol (26.5 ppm) to rats significantly increased muscle weight by 15%. In vitro, zinterol stimulated lactate release from L8 cells whereas propranolol inhibited this process, demonstrating that these cells have functional beta-receptors both before and after fusion. We measured several anabolic processes, in both serum stimulated and quiescent cells, over a wide range of zinterol concentrations. Zinterol had no effect on protein or DNA synthesis, protein degradation, or rates of amino acid uptake. These data suggest that the in vivo muscle growth stimulation is either indirect or some in vivo requirements (e.g. tension and nerve interactions) are necessary for expression of the effect. PMID- 2567599 TI - Lactic acidosis in theophylline overdose. AB - An 18-year-old man with theophylline overdose developed an increased anion gap metabolic acidosis. Serum lactate levels were markedly elevated. A direct correlation was found between the increasing theophylline level, clinical hyperadrenergic state, and the worsening acidosis. Early hemoperfusion reversed the acidosis, the elevated serum theophylline level, and the hyperadrenergic state. This case substantiates the role of lactate accumulation in the metabolic acidosis associated with isolated theophylline toxicity. PMID- 2567600 TI - Cyanide poisoning successfully treated without 'therapeutic methemoglobin levels'. AB - A 24-year-old woman ingested an unknown amount of potassium cyanide in a suicide attempt. Coma and metabolic acidosis developed. Administration of the Lilly Cyanide Antidote kit (Eli Lilly and Co, Indianapolis) resulted in prompt resolution of symptoms and full recovery. Whole blood cyanide level was 13 micrograms/mL approximately one hour after ingestion. The highest measured methemoglobin level after sodium nitrite administration was 9.2%, demonstrating that attaining a "therapeutic methemoglobin level" of 25% is unnecessary to insure a satisfactory clinical outcome. Because severe hypotension or excessive methemoglobinemia can be caused by the sodium nitrite component of the Lilly kit, only enough to produce an acceptable clinical response should be administered. PMID- 2567598 TI - Association of complement alleles C4AQ0 and C4B5 with rheumatoid arthritis in Japanese patients. AB - We investigated polymorphisms of complement components C2, C4, and factor B (BF) in Japanese patients with rheumatoid arthritis (RA). The frequencies of C4AQ0 (32.1%) and C4B5 (35.9%) among RA patients were significantly higher than among healthy control subjects. C4B5 was strongly associated with HLA-Bw54, Bw59, DR4.1, and DQw4. C4AQ0 showed no association with HLA-Bw54 or Bw59, but there was weak association with HLA-DR4.1 and DQw4. The number of persons with both C4AQ0 and C4B5 was significantly higher in the RA patient group (relative risk 13.5). C2C and BFS were the most common alleles in RA patients, as well as in healthy control subjects. These data support the existence of 2 different putative susceptibility haplotypes (HLA-Bw54 or Bw59;C2C; BFS;C4A3;C4B5;DR4.1;DQw4 and C2C;BFS; C4AQ0;C4B1 or C4B2) in Japanese patients with RA. PMID- 2567601 TI - [Treatment of solar urticaria: advantage of terfenadine]. AB - Solar urticaria is a rare photodermatosis, but it is extremely incapacitating and therapy is usually ineffective. Three patients who took a daily dose of 240 mg terfenadine were able to be normally exposed to sunlight. This efficacy was confirmed by photobiological tests. The minimal dose necessary to induce urticaria was increased at least three times. These results confirming recently published ones. Terfenadine at 240 mg per day seems to be the treatment of choice for solar urticaria because of its efficacy and excellent tolerance-especially as there is no sedative effect. PMID- 2567602 TI - Synthesis of 4% Glu-containing Val1 and Ile1-polypentapeptides: model protein systems for demonstrating mechanochemical coupling. AB - The synthesis of 4% Glu-polypentapeptide (PPP) (i.e., 4 Glu residues per 100 amino acid residues) and 4% Glu-Ile1-PPP, in which Val1 is substituted by a more hydrophobic Ile residue, is carried out by copolymerizing the p-nitrophenyl active esters of GE(OMe)GVP and GE(OMe)GIP with their parent pentamers GVGVP and GVGIP in 1:4 ratios, respectively. After removal of the methyl ester on the side chain of Glu, these polymers exhibited a remarkable pH dependence of the temperature for their inverse temperature transitions, which are followed as turbidity development at 300 nm. On gamma-irradiation crosslinking, the elastomeric bands obtained exhibited a pH-mediated contraction and relaxation. Thus, for the first time, mechanochemical coupling is demonstrated in a synthetic polypeptide system. That the basic mechanism involves the hydrophobic effect (chemical modulation of an inverse temperature transition) and not ion-ion electrostatic repulsion is also discussed. PMID- 2567603 TI - Effect of DSP4, a neurotoxic agent, on attentive behaviour and related electrocortical activity in cat. AB - Six behaving cats were administered N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP4) (i.p. 10 mg/kg), a neurotoxic agent known to destroy noradrenergic endings in the CNS. They were placed, both before (control) and after treatment in 3 different situations, each time for 90 min: (1) a 'neutral' one (N) with no significant stimuli; (2) another one eliciting focused attention (F); (3) a third one, creating a situation of 'expectancy of an event to occur' (E), with the animal usually displaying a posture of quiet waking. Simultaneously, the electrocorticogram (ECoG) was recorded from the sensorimotor and parietal cortex through implanted electrodes. We already knew and were able to confirm that the normal animals (i.e. before treatment) displayed distinct behavioural patterns depending on the situation and distinct accompanying parietofrontal ECoG activities, with a dominance of drowsiness and sleep during N, that of a 36-Hz 'beta' rhythms in condition F, and of 14-Hz 'mu' rhythms in condition E. It was shown that the prevailing attitude of the animals after treatment was now, in all 3 situations, that of 'quiet waking and/or expectancy-like watching', with a large if not exclusive dominance of only one ECoG pattern, namely mu. These changes were considered as due to a release of the mu system from a noradrenergic modulatory blockade, in accordance with some of our previous data. An immunohistochemical study with anti-tyrosine hydroxylase antibody was also performed; it confirmed that after DSP4 treatment there were substantial alterations in the immunoreactivity of locus coeruleus cells, the structure which is likely to be involved in this NAergic control of the mu rhythms and of its concomitant behaviour. PMID- 2567605 TI - [The occurrence of tyrosine hydroxylase-immunoreactive neurons in a parietal lobe ganglioglioma]. AB - A parietal lobe ganglioglioma in a 61-year-old male was investigated ultrastructurally and immunohistochemically, using antiserum against tyrosine hydroxylase (TH), a rate-limiting enzyme of the catecholamine (CA)-synthesizing pathway. On light microscopy, the tumor was composed mainly of neuronal and astrocytic cells. The neuronal cells had typical round vesicular nuclei with prominent nucleoli and showed abundant cytoplasm with variable amounts of Nissl bodies. Bi-nucleated neuronal cells were occasionally found. The astrocytic cells had eosinophilic cytoplasm of variable shapes and hyperchromatic small nuclei. Electron microscopy revealed numerous dense core vesicles (DCVs) in the neuronal cytoplasm and processes. Immunohistochemically, TH-immunoreactive neuronal cells were found sporadically in the tumor tissue. They were often binucleated or multinucleated. The presence of TH-immunoreactive neuronal cells in the present case is of interest in considering the origin of neuronal cells in ganglioglioma. Some investigators have considered that they may arise from the ectopic autonomic neuronal cell nests in view of the presence of DCVs. However, Gasper et al. have recently demonstrated the presence of TH-immunoreactive neurons in the normal human cerebral cortex. Therefore, it may also be possible to consider that TH immunoreactive neuronal cells in this tumor originated from a certain dysgenetic focus containing such TH-immunoreactive neurons in the developmental stage. PMID- 2567604 TI - Attenuation of the effects of ethanol on social behavior by alpha 2-adrenoceptor antagonists. AB - In a social behavior test in mice ethanol (2 g/kg) significantly reduces the time spent in social interaction and increases locomotor activity. The relatively selective alpha 2-adrenoceptor antagonists atipamezole and idazoxan alone have no significant effect on either social interaction or locomotor activity in this test. However, when coadministered with 2 g/kg ethanol, atipamezole (0.3-3.0 mg/kg) significantly attenuated the ethanol-induced reduction in the time spent in social interaction without any effect on ethanol's locomotor stimulant action. Idazoxan (0.3-1.0 mg/kg) also showed a trend towards reversing the reduction in the time spent in social interaction although this was accompanied by a significant attenuation of the locomotor stimulant effect of ethanol. The results indicate that alpha 2-adrenoceptors may modulate ethanol's effects on social interaction in mice. PMID- 2567606 TI - The immediate or delayed replacement of teeth by permucosal intra-osseous implants: the Tubingen implant system. Part 1. Implant design, rationale for use and pre-operative assessment. AB - No single implant system is applicable to all clinical problems which may be treated with implants. Clinicians who wish to develop expertise in the field of dental implantology should decide which clinical situations they wish to treat with implants and then receive an appropriate level of instruction in the use of systems which have been shown, in refereed literature, to be successful in the treatment of such problems. The Tubingen implant system has been in clinical use for more than 13 years, following preliminary basic research and animal studies, and is supported by an extensive literature. It is applicable primarily to the immediate or delayed replacement of individual teeth which are subsequently to be restored with a post crown. It may also be used in other situations, such as in combination with bridge restorations. This is the first of two papers which describe in detail the range of applications, patient selection and assessment, and the surgical and restorative techniques involved in its use. PMID- 2567607 TI - P-glycoprotein gene amplification and expression in multidrug-resistant murine P388 and B16 cell lines. AB - P-glycoprotein gene (mdrl) amplification and expression were examined in murine leukaemia P388/DX and melanoma B16VDXR cell lines, which exhibit a high level of resistance to a selecting agent, doxorubicin, and express a multidrug-resistant phenotype because they are cross-resistant to multiple cytotoxic drugs. The multidrug-resistant phenotype was obtained in different conditions of selection (in vivo and in vitro for P388/DX and B16VDXR, respectively). In both multidrug resistant cell lines, an increased expression of P-glycoprotein gene (5 kb transcript detected in Northern blots) was observed and the level of P glycoprotein mRNA correlated with the degree of resistance. In addition, high molecular weight mRNAs homologous to mdrl gene sequence were consistently detected only in P388/DX cells. Overexpression was associated with a high level of gene amplification only in resistant melanoma cells, whereas it occurred in P388/DX cells with a marginal increase in gene copy number. These results, suggesting that different genetic mechanisms could be responsible for P glycoprotein overexpression, emphasise the complexity of genetic regulation that may affect tumour cell sensitivity to cytotoxic agents. PMID- 2567608 TI - Synthesis of somatostatin by breast cancer cells and their inhibition by exogenous somatostatin and sandostatin. AB - Three human breast cancer cell lines ZR-75-1, MDA-MB-436 and MCF-7 were found to contain respectively, 3.06, 2.69 and 1.86 fmol of somatostatin-like immunoreactivity (SLI) per 10(6) cells. Since SLI is undetectable in the passaging media it must, therefore, be synthesised by the cells. In the presence of fetal calf serum the cells were growth inhibited by addition of somatostatin or its long-lasting analogue, Sandostatin, but only after 3 days of continuous exposure. A 1-day exposure to either peptide had little or no effect on subsequent cell growth in peptide-free medium. Inhibition of cell proliferation is not due to cytotoxic effects of the dose used (500 ng ml-1, each) since both peptides caused short-term stimulation of growth in the absence of serum. PMID- 2567609 TI - Muscarinic receptor subclassification and G-proteins: significance for lithium action in affective disorders and for the treatment of the extrapyramidal side effects of neuroleptics. AB - The classification of muscarinic receptors into M1 and M2 subtypes and the involvement of guanine nucleotide binding proteins (G-proteins) as major mediators of receptor information transduction in the cholinergic and other neurotransmitter systems have prompted us to undertake studies both at receptor and postreceptor levels that may shed light on the importance of these new findings to the pharmacotherapy of manic-depressive illness and of extrapyramidal syndromes. We searched for patterns of muscarinic selectivity among the commonly used anticholinergics (biperiden, procyclidine, trihexyphenidyl, benztropine, and methixen) through radioligand receptor studies in various rat tissues. The drugs showed a range of selectivity, from the totally nonselective methixen to the highly M1-selective biperiden. Sinus arrhythmia measurements were undertaken in psychiatric patients treated with different antiparkinsonian anticholinergics. The extent of sinus arrhythmia suppression was inversely correlated with the degree of M1 selectivity of the drugs used, advocating the use of M1-selective antiparkinsonian anticholinergics like biperiden in the treatment of extrapyramidal side effects. The implications of muscarinic receptor subclassification were further extended to include postreceptor phenomena. We have directly studied G-protein function by measuring cholinergic agonist-induced increases in guanosine triphosphate (GTP) binding to these proteins. This cholinergic agonistic effect was shown to be exerted by G-proteins other than Gs (the adenylate cyclase stimulatory G-protein), i.e., Gi (the adenylate cyclase inhibitory G-protein) or Gp [the G-protein activating phosphatidylinositol (PI) turnover], as ribosylation by pertussis toxin abolished this cholinergic effect, whereas it was unaffected by cholera toxin. Pertussis toxin-blockable, carbamylcholine-induced increases in GTP binding capacity were found to be mediated through M1 muscarinic receptors, as M1-selective antagonists were 100 fold more effective than M2 selective antagonists in blocking carbamylcholine effects. Moreover, carbamylcholine effect was exclusively detected in tissues predominantly populated by M1 receptors. Our results thus suggest that carbamylcholine-induced increases in GTP binding are exerted through M1 receptors interacting with Gp. At therapeutically efficacious concentrations, lithium completely blocked carbamylcholine-induced increases in GTP binding capacity in both in vitro and in vivo experiments.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2567612 TI - Melanocyte-stimulating hormone in tardive dyskinesia. PMID- 2567611 TI - Failure of clonidine in tardive dystonia: implications for nosology. PMID- 2567610 TI - Effect of neuroleptic withdrawal on plasma prolactin: a possible marker of receptor adaptation. AB - Stable schizophrenic and schizoaffective outpatients underwent abrupt withdrawal from a fixed dose of neuroleptic. Prolactin concentrations were determined preceding and following drug withdrawal. Basal prolactin concentrations were significantly lower 3, 4, or 5 days following drug withdrawal than on subsequent days. This rebound effect may be related to dopaminergic receptor changes in the tuberoinfundibular system that are induced by chronic neuroleptic administration. PMID- 2567613 TI - R-R interval during neuroleptic treatment. PMID- 2567614 TI - Antimycobacterial chemotherapy in inflammatory bowel disease. AB - We report on a case, ulcerative colitis and another of Crohn's disease. During a relapse which was unresponsive to conventional therapy, acid-fast bacilli were found in colonic biopsies. Conventional therapy was substituted with antimycobacterial chemotherapy (rifampicin, isoniazid and ethambutol) which was responsible for a marked improvement. However, a relapse occurred during chemotherapy and no acid-fast bacilli were found. The patients became responsive to sulphasalazine and corticosteroid therapy once again. It appears that Mycobacteria play a collateral role in inflammatory bowel disease and that once they have been eliminated the original disease re-emerges. PMID- 2567615 TI - [Elimination of disorders of the electrical stability of the heart in experimental myocardial infarct and postinfarct cardiosclerosis under the influence of a benzodiazepine receptor agonist]. AB - It was shown on Wistar male rats that agonist of benzodiazepine receptors phenazepam considerably suppressed the heart ectopic activity and eliminated disturbances of the heart electric stability in acute myocardial infarction and postinfarction cardiosclerosis. The drug prevented to a considerable extent the death rate of animals within the first day following the ligation of coronary artery and the fall of arterial pressure in animals with the acute myocardial infarction. Possible mechanism of the benzodiazepine agonist effect is under discussion. PMID- 2567616 TI - [Phenotypic and functional characteristics of the subpopulations of regulatory T lymphocytes participating in the stimulation of hemopoiesis during stress]. AB - The phenotypic and functional characteristics of T-lymphocytes detected in bone marrow at stress was studied in experiments on mice. The T-lymphocytes-regulators taking part in hemopoiesis stimulation on 5-7 days after 10-hours immobilization have phenotype by Lyt 1+, 2+, L3T4-. PMID- 2567617 TI - Gallstone lithotripsy with the pulsed dye laser: in vitro studies. AB - The use of a pulsed dye laser to fragment human gallstones is reported. Laser energy was delivered by a 320 microns quartz fibre at wavelengths of 440, 480, 504, 560, 590, and 635 nm, in 1 microsecond pulses at energies of 5-60 mJ per pulse, producing peak powers of the order of 10(5) W. Cholesterol, pigment and mixed stones were fragmented. The shorter wavelengths were most efficient, using significantly less energy at 440 and 480 nm than at the longer wavelengths. Pigment stones were fragmented with significantly less energy than those in which cholesterol predominated. Thermal imaging during laser treatment showed no significant rise in temperature in the bulk of the stone. Absorption spectroscopy of bile revealed a peak at 450 nm, coinciding with the known absorption characteristic of bilirubin. Bile strongly attenuated light at shorter wavelengths, even at dilutions as great as 1:20 and protected samples of biliary epithelium from laser damage except when the optical fibre was in direct contact. The pulsed dye laser may offer significant advantages over current methods of biliary stone fragmentation. PMID- 2567618 TI - Relationships between acid-base balance, serum composition and colostrum absorption in newborn calves. AB - Twenty-seven newborn Holstein bull calves were bottle-fed 2 litres of pooled colostrum which had been stored at -20 degrees C. Blood gas analysis before feeding showed a partially compensated respiratory acidosis in most of the calves, although they all appeared to be clinically normal. Mean venous blood pH was 7.346, carbon dioxide tension (PCO2) was 57.5 mmHg (7.6 kPa), bicarbonate was 30.6 mmol/l and base excess was 3.82 mmol/l. Mean serum IgG1 increased to 8.1 g/l after feeding colostrum. Several significant positive correlations were observed between post-absorptive serum protein, IgG1, IgM, gamma-glutamyltransferase (gamma GT) and D-xylose. Calves with either low serum albumin, high serum CK or low serum gamma GT before feeding tended to have less absorption of colostral protein. It was concluded that reduced absorption of IgG1 from colostrum is associated with hypercapnia in apparently healthy newborn calves. PMID- 2567619 TI - Immunoregulation of murine plasmacytoma. I. Generation of anomalous killer cells in vitro by cocultivation with MOPC 104E. AB - Murine plasmacytoma MOPC 104E-K181 is a tissue culture cell line of MOPC 104E derived from BALB/c mice. MOPC 104E-K181 implanted subcutaneously in syngeneic normal mice regresses spontaneously after an initial growth of about 10 mm. Mice that regressed tumors or mice immunized intraperitoneally with mitomycin C treated MOPC 104E-K181 myeloma could reject subsequent challenge of viable K181 myeloma cells. In contrast to euthymic mice, T-cell-deficient athymic nude mice developed subcutaneous tumors after challenge and died from progressive tumor growth, suggesting the critical role of T cells in tumor regression. In vitro induction of cytotoxic cells was used to define the immunologic mechanism by which the host can suppress tumor growth. Spleen cells from immune mice did not show cytolytic activity in 51Cr release cytotoxicity assay, but showed inhibitory action of tumor proliferation in vitro at an effector cell to target cell ratio of 500:1 in a [3H]thymidine incorporation assay. To determine if cytotoxicity could be induced against MOPC 104E-K181 cells, in vitro sensitizing cultures were studied. We have demonstrated that normal BALB/c spleen cells became cytotoxic against MOPC 104E-K181 cells after 5 days cultivation with mitomycin C-treated stimulator cells at an optimal responder to stimulator cell ratio of 5:1. Treatment of anti-Thy-1.2 serum plus complement abolished cytotoxic activity of effector cells. Cytotoxic cells lysed not only MOPC 104E-K181 cells used for stimulation but also H-2k osteosarcoma cells. It was concluded that Thy-1.2 positive cytotoxic cells with nonspecific anomalous reactivity could be induced in murine plasmacytoma-stimulating cultures. PMID- 2567620 TI - Metachromatic leukodystrophy (MLD) in hospitalized adult schizophrenic patients resistant to drug treatment. AB - Metachromatic leukodystrophy (MLD) is a rare inherited neurodegenerative disease associated with a defect in the catabolism of sulphatide (galactocerebroside sulphate) which accumulates in the nervous system. MLD can be diagnosed biochemically by demonstrating deficiency in the activity of the enzyme arylsulphatase A (ASA) and an excess of sulphatide in urine and tissues. Clinically adult MLD may present as a schizophrenic-like psychosis, which typically develops years before the onset of neurologial signs which are not inevitable. Urinary ASA was investigated in 99 chronic hospitalized psychiatric patients (including 77 schizophrenics). Thirteen showed reduced ASA activity. Of the nine who were available for further study, only one evinced reduced ASA activity in other tissues (for example, leukocytes and cultured fibroblasts). However, there was no evidence of sulphatidurea with impaired sulphatide hydrolysis in his intact cultured fibroblasts. Therefore, he tested negative for MLD. This biochemical profile is known as pseudosulphatase deficiency. The possible relationship of pseudosulphatase deficiency to schizophrenic-like conditions is discussed. PMID- 2567621 TI - Pharmacotherapy of the borderline patient: a critical review and clinical guidelines. AB - The literature on the pharmacotherapy of Borderline Personality Disorder (BPD) is critically reviewed, and suggestions for the appropriate clinical use of psychotropic agents and directions for future research are made. PMID- 2567622 TI - Rat liver gamma glutamyl transpeptidase mRNA differs in the 5' untranslated sequence from the corresponding kidney mRNA. AB - A clone containing the entire protein-coding region of gamma glutamyl transpeptidase (GGT) from ethoxyquin- (EQ) treated rat liver has been isolated from a cDNA library. DNA sequence analysis showed that the protein-coding and 3' untranslated regions are almost identical to rat kidney GGT, but the sequences differ in the 5' untranslated region. Southern blot data suggest a single gene for GGT in the rat, hence the different mRNAs may originate by differential transcriptional and/or posttranscriptional processing. PMID- 2567623 TI - Effects of bryostatins and retinoic acid on phorbol ester- and diacylglycerol induced squamous differentiation in human tracheobronchial epithelial cells. AB - Previous studies have shown that normal human tracheobronchial epithelial (HBE) cells undergo squamous differentiation upon treatment with phorbol 12-myristate 13-acetate (PMA). In this study, we report that induction of this differentiation program is accompanied by an increase in the accumulation of cholesterol sulfate and in transglutaminase type I activity, two markers of squamous differentiation. Several carcinoma cell lines did not exhibit an increase in these differentiation markers after PMA-treatment and appear to have acquired a defect in the mechanism that triggers differentiation. The diacylglycerol analogue, didecanoylglycerol (diC10), was also able to induce squamous differentiation. Bryostatin 1, another activator of protein kinase C, did not induce terminal cell division or increase cholesterol sulfate accumulation or transglutaminase type I activity. Bryostatin 1 not only failed to inhibit cell proliferation and to induce differentiation but antagonized the PMA- and diC10-induced commitment to terminal differentiation. The bryostatin blocked both the PMA-induced terminal cell division as well as the expression of the two differentiation markers. Retinoids were found not to affect the PMA-induced commitment to terminal cell division but did inhibit the expression of the differentiated phenotype. Our results indicate that the bryostatins and retinoids affect the multistep process of squamous differentiation in tracheobronchial epithelial cells at two different stages. PMID- 2567624 TI - [Secondary prevention in the post-infarction period]. PMID- 2567626 TI - [Preliminary study on epidemic hemorrhagic fever (EHF) in Guizhou Province]. AB - During 1962-1986, EHF cases have been reported in 50 out of 82 counties in Guizhou province. It was high in northern Guizhou province. However, serological survey of the population showed that endemic area extended to 51 counties. The inapparent infection rate ranged from 0 to 4.04%. In 76 out of 82 counties the rodents were found to carry hantavirus. Apodemus agrarius of Zunyi county of Guizhou and Rattus norvegicus of the eastern and south Guizhou are important reservoirs of EHF. Hantavirus has been isolated both from patients and from rodents. PMID- 2567627 TI - [Studies on inapparent infection of epidemic hemorrhagic fever in population of Shanxi Province]. AB - Detection for IgG antibody to virus of epidemic hemorrhagic fever (EHF) in sera from healthy people in Shanxi province were carried out by IFAT. Two hundred thirty eight of 4873 serum samples were antibody-positive. Total positive rate was 4.88%. The inapparent infection rate of EHF in residents of endemic areas was 5.48%. There was a statistically significant difference of epidemic strength among different counties. Based on the analysis of antibody titers of positive sera, value of upper limit (P95) of antibody titre for inapparent infection was 1:315. It was suggested that the antibody titre of 1:320 by IFAT in a single sample of serum from patient with clinical manifestations of EHF might be used as a criteria for specific diagnosis of EHF. PMID- 2567628 TI - [Preliminary study on the virulent difference between viral strains isolated from Rattus norvegicus and Apodemus agrarius in Fujian Province]. AB - Virulence between virus strains of hemorrhagic fever with renal syndrome (HFRS) from R. norvegicus and A. agrarius were different, besides the diversity in their epidemiological feature, clinical symptoms and antigenicity. Virulence of the virus strains from R. norvegicus, especially those from nonepidemic areas, were weak than those from A. agrarius. Sucking mice might be used for determining virulence of the virus strains from various source. Foci of HFRS had been in southern Fujian, but human cases of HFRS had rarely been found, or only a few mild cases were found in a few areas. This might be related to weaker virulence of the virus strains from R. norvegicus. The prevalent degree of the disease might be related to HFRSV of wild rattus type in the area. PMID- 2567625 TI - Differential expression of tissue transglutaminase in human cells. An immunohistochemical study. AB - Tissue transglutaminase is an intracellular enzyme without established physiological function. Biochemically it can be detected in all organs, but no systematic in situ localization has been carried out so far. Here we report the immunohistochemical localization of transglutaminase in human tissues using an affinity purified, monospecific anti-human transglutaminase antibody. It is shown that the widespread organ distribution of the enzyme is the consequence of its occurrence in ubiquitous cell types such as endothelium and smooth muscle cells. Some organ-specific cell types express the enzyme constitutively (mesangial cells, renomedullary interstitial cells, thymic subcapsular epithelium, colonic pericryptal fibroblasts), while in others it seems to be induced either by external stimuli (epithelium of the female breast) or as part of their differentiation/maturation program (developing nephrons, enterocytes of the small intestine). The presence of tissue transglutaminase can be demonstrated in derivatives of all germ layers and in the trophoblast. The functional implications of these findings are presently unknown; however, based on its distribution the role of this enzyme in compartmentation and preservation of tissue integrity against stress may be suggested. PMID- 2567629 TI - [Analysis on distribution of epidemic hemorrhagic fever in different area]. AB - The data showed that there was acrrelation between EHF and host animals as well as types of natural foci in plains, hills and mountain areas of Tian Tai county. It was clear that the main host animals were R. norvegicus and A. agrarius in plain areas, so there are mixture foci of R. norvegicus and A. agrarius in the above areas. The morbidity of EHF and the positive rate of mouse lung antigen as well as the inapparent infections rate of healthy population were high in plains. As the attitude elevated, it gradually formed a natural focus of R. norvegicus as the main host animal in hills and mountaim areas. The positive rate of the mouse lung antigen diminished sharply. Both the incidence rate of EHF and the inapparent infection rate of healthy population were lower. PMID- 2567630 TI - [Study on geographic epidemiology of epidemic hemorrhagic fever (EHF) in China]. AB - The distribution, epidemic intensity and the focus type of EHF at county's level in China was studied by the method of discriptive, serological and geographic epidemiology. The EHF foci distributed in 1,257 counties within 25 provinces (cities, autonomous regions) by 1986, 633 of them were found before 1980 and the other 624 after it. The epidemic area have extended nearly twice in the last years. The extension of rattus type focus was most significant. The number of focus of Rattus type increased from 20 counties in 1980 (account for 3.73% of total foci) to 288 in 1986 (26.77%); the mixed type also enlarged from 98 (18.28%) to 267 (24.81%); while the Apodemus type was relatively steady. More than 250,000 small animals were captured and tested in this research. 4 orders 8 families and 37 species were found carrying EHF virus antigen. The research pointed out that the main kinds of the reservoir were limited. In addition, the animals which carry EHF virus were found also in 111 non-case-reported counties, so these areas could be a potential EHF's foci. The relationship between the distribution of EHF focus (especially Apodemus type) and natural geographic factors, such as the physical features of a place, precipitation and water system etc, was analysed. The preventive strategies and suggestions were also discussed. PMID- 2567632 TI - Transcriptional activation and repression by Ultrabithorax proteins in cultured Drosophila cells. AB - Homeotic genes of Drosophila melanogaster such as Ultrabithorax (Ubx) and Antennapedia (Antp) have long been thought to select metameric identity during development by controlling the expression of various target genes. Here we describe a cotransfection assay in cultured D. melanogaster cells that is used to demonstrate that Ubx proteins (UBX) can repress an Antp promoter fusion and activate a Ubx promoter fusion, activities predicted from genetic studies. We show (a) that UBX proteins regulated the level of accurately initiated Antp P1 and Ubx transcripts, (b) that activation of the Ubx promoter required a downstream cluster of UBX binding sites, and (c) that binding site sequences were sufficient to confer regulation on a heterologous promoter, regardless of their orientation or precise position. We conclude that UBX proteins are transcriptional repressors and activators, and that their actions are mediated by binding to promoter region sequences. Each member of the UBX protein family has similar regulatory abilities, but the properties of synthetic mutant forms suggest that UBX proteins may have a modular design similar to other transcriptional regulators. PMID- 2567631 TI - Transcriptional activation by the Antennapedia and fushi tarazu proteins in cultured Drosophila cells. AB - Drosophila homeodomain proteins bind to specific DNA sequences in vitro and are hypothesized to regulate the transcription of other genes during development. Using a cotransfection assay, we have shown that homeodomain proteins encoded by the homeotic gene Antennapedia (Antp) and the segmentation gene fushi tarazu, as well as a hybrid homeodomain protein, are activators of transcription from specific promoters in cultured Drosophila cells. Sequences downstream of the Antp P1 and Ultrabithorax transcription start sites mediate the observed activation. A TAA-rich DNA sequence to which the Antp protein binds in vitro is sufficient to confer regulation on a heterologous promoter. The results demonstrate that homeodomain proteins are transcriptional regulators in vivo and that in cultured cells, different homeodomain-containing proteins can act upon a common sequence to modulate gene transcription. PMID- 2567633 TI - Gene activation and DNA binding by Drosophila Ubx and abd-A proteins. AB - The Ubx and abd-A gene products are required for proper development of thoracic and abdominal structures in Drosophila. We expressed LexA-Ubx and LexA-abdA fusion proteins in yeast. These proteins activated expression of target genes that carried either upstream LexA operators or upstream Ubx binding sites. Both proteins contain homeodomains. Experiments with mutant fusion proteins show that the homeodomain is not required for the proteins to form dimers or enter the nucleus, and that, when DNA binding is provided by the LexA moiety, the homeodomain is not required for gene activation. Our results suggest that the homeodomain is necessary for these proteins to bind Ubx sites, but that the homeodomain does not contact DNA exactly like bacterial helix-turn-helix proteins. Finally, our data suggest that gene activation by these proteins is a simple consequence of their binding to DNA, while negative gene regulation requires that these proteins act together with other Drosophila gene products. PMID- 2567634 TI - Stochastic appearance of mammary tumors in transgenic mice carrying the MMTV/c neu oncogene. AB - Transgenic mice carrying the activated c-neu oncogene under the control of the mouse mammary tumor virus (MMTV) long terminal repeat were produced. Epithelial hyperplasia of epididymis, seminal vesicles, and salivary glands, and dysplasia of harderian glands, were induced. Moreover, in females of our four lines, independent but multiple mammary tumors arose asynchronously, between 5 and 10 months of age, as stochastic events. Histologically, poorly differentiated adenocarcinomas, with intratumor necrosis and calcifications, arose adjacent to morphologically normal epithelium. High transgene expression was detected in all mammary tumors tested and in normal mammary glands before the appearance of the tumors. Together these results suggest that the expression of the activated c-neu oncogene was necessary but not sufficient to induce malignant transformation of the mammary epithelial cells. These tumors appear to be an adequate model for human breast cancers overexpressing c-neu. PMID- 2567635 TI - Mix.1, a homeobox mRNA inducible by mesoderm inducers, is expressed mostly in the presumptive endodermal cells of Xenopus embryos. AB - In frogs, mesoderm presumably derives from presumptive ectoderm by induction under the control of diffusible substances produced by the endoderm. To analyze the early phase of mesoderm induction, I have isolated cDNA copies of mRNAs induced in presumptive ectoderm by mesoderm inducing factor secreted by the XTC cell line. One of the inducible mRNAs encodes a homeodomain-containing protein that is likely to play a regulatory role in development. Mix.1 behaves as an immediate early response to induction, and its kinetics of expression suggest a major role for MBT in the control of inducible gene expression. Unexpectedly, Mix.1 is expressed mostly in the future endoderm, suggesting that endoderm may be formed by induction in a similar way as mesoderm. PMID- 2567636 TI - A susceptibility locus for multiple sclerosis is linked to the T cell receptor beta chain complex. AB - Inheritance of T cell receptor beta chain (TCR beta) genes was analyzed in families of 40 sibling pairs concordant for the relapsing-remitting form of multiple sclerosis (MS). TCR beta haplotypes were determined by segregation analysis of polymorphic markers within the TCR beta complex. The mean proportion of TCR beta haplotypes identical by descent (IBD) inherited by MS sibling pairs was significantly increased compared with expected values (means test, p less than 0.004), whereas the distribution of haplotype sharing was random when MS patients were compared with their unaffected siblings. Furthermore, one allelic form of a TCR beta variable region gene segment was overrepresented on MS chromosomes compared with those parental chromosomes not transmitted to MS offspring both in the MS sibling pair families and in a second group of families containing only one individual affected with MS. These results demonstrate that a gene within the TCR beta complex or a closely linked locus influences susceptibility to MS. PMID- 2567637 TI - The gradient morphogen bicoid is a concentration-dependent transcriptional activator. AB - The bicoid (bcd) protein is expressed in an anteroposterior gradient in early Drosophila embryos and controls the zygotic activation of the segmentation gene hunchback (hb) in a broad but precisely bounded anterior domain. Here we show that the hb gene contains multiple regulatory elements that mediate transcriptional activation in response to bcd protein. Further, we demonstrate that the resulting patterns of expression in vivo depend critically on both the bcd gradient profile and the number and quality of these hb elements. Finally, we show that these same elements mediate bcd-dependent transcriptional activation in yeast and that this interaction requires distinct DNA binding and activating regions in the bcd protein. Our results argue that bcd protein normally binds and activates the hb gene in a concentration-dependent fashion, thereby allowing the gradient of bcd protein to dictate where the hb gene is initially turned on in early embryos. They also suggest that the bcd gradient has the instructive capacity to activate other subordinate control genes by the same mechanism, each in a distinct spatial domain according to its affinity for bcd protein. PMID- 2567638 TI - [The changes in beta-adrenergic receptors and cyclic AMP levels of lymphocytes in patients with COPD]. AB - The maximal binding capacity (Bmax) of beta-adrenergic receptors and the production of intracellular cAMP in response to isoproterenol in peripheral blood lymphocytes were measured by radioligand binding assay and competitive protein binding method respectively. Two groups of COPD patients (19 cases was at the onset of disease, 17 cases was stable) were investigated and 20 normal subjects as control. The amount of Bmax at the onset of COPD (208.55 +/- 14.14fmol/10(8)cells) and during the stable stage (299.25 +/- 13.90fmol/10(6)cells) was significantly lower than that in normal subjects (338.35 +/- 12.22fmol/10(8)cells) (P less than 0.01). There were also obvious differences in amounts of Bmax between two groups of COPD (P less than 0.01). The changes of intracellular cAMP were similar to the results of Bmax. This study indicated that beta-adrenergic receptor function is attenuated in COPD patients especially at the onset of the disease. The possible mechanism of the reduction of beta-adrenergic receptor and the clinical significance in the use of beta adrenoceptor agonist were discussed. PMID- 2567640 TI - Time-series analysis of heart rate variability during submaximal exercise. Evidence for reduced cardiac vagal tone in animals susceptible to ventricular fibrillation. AB - Periodic fluctuations in the R-R interval have been used as noninvasive measures of cardiac autonomic tone. For example, a reduced heart rate variability has been shown to correlate with an increased mortality in patients recovering from myocardial infarction. The effects that physiologic perturbations such as exercise have on this heart rate variability have not been investigated. Therefore, heart rate variability was measured throughout a submaximal exercise test in 36 mongrel dogs with healed anterior myocardial infarctions. The amplitude of the respiratory component (0.24-1.04 Hz) was determined by time series analysis techniques and was used as an index of cardiac vagal tone. On a subsequent day, a 2-minute coronary occlusion was initiated during the last minute of exercise. Twenty-two animals developed ventricular fibrillation (susceptible), whereas 14 animals did not (resistant). Exercise elicited a significantly greater increase in heart rate (resistant, 205.4 +/- 7.1; susceptible, 227.0 +/- 5.4 beats/min) in susceptible animals, which was accompanied by a greater reduction in the cardiac vagal tone index (resistant, 2.7 +/- 0.3; susceptible, 1.1 +/- 0.2 ln msec2) as compared with resistant animals. Conversely, atropine sulfate (50 micrograms/kg) given during exercise elicited a greater heart rate increase in the resistant dogs (heart rate change: resistant, 54.2 +/- 7.0; susceptible, 18.7 +/- 4.4 beats/min). Taken together, these data suggest that exercise elicited a greater reduction in cardiac vagal tone in animals known to be susceptible to ventricular fibrillation. PMID- 2567639 TI - Enhancement of hepatocarcinogenesis in rats by dietary fructose. AB - The effects of oral fructose on hepatocarcinogenesis were investigated with cytomorphological, cytochemical and stereological methods. Carcinogenesis was induced in male Sprague-Dawley rats by application of N-nitrosomorpholine (NNM) for 7 weeks. Afterwards, the animals received fructose in the drinking water (120 g/l) and food ad libitum (group I) or tap water and food ad libitum (group II). The incidence of hepatocellular carcinoma in rats treated with NNM plus fructose was 46% as compared to 24% in animals receiving NNM alone (P less than 0.05). There was no difference in the incidences of other malignancies between the groups (group I: 32.1%, group II: 32.0%). Morphometric evaluation of preneoplastic liver lesions indicated the enhancing effect of the fructose treatment several months before malignant tumors appeared. As early as 6 weeks after treatment the hepatic parenchyma occupied by focal lesions was increased from 6.7% in the animals which had received NNM alone to 8.5% (P less than 0.05) in animals having received NNM plus fructose. This increase was predominantly caused by an increase in glycogen storing foci (P less than 0.0005). In addition, the fructose treatment caused a histochemically detectable increase in the activity of glucose-6-phosphatase and glucose-6-phosphate dehydrogenase in both the hepatocytes of the focal lesions and the surrounding parenchyma. In the NNM plus fructose group the activity of the glucose-6-phosphatase in the foci was frequently approximately equal to the activity in the parenchyma of untreated controls. The striking increase in the activity of this enzyme in the surrounding hepatocytes, however, still sharply demarcated the lesions. The potential mechanisms by which fructose enhances hepatocarcinogenesis are discussed. PMID- 2567641 TI - Postnatal changes in glutamate stimulated phosphoinositide turnover in rat neocortical synaptoneurosomes. AB - Glutamate was found to stimulate the accumulation of [3H]inositol phosphate in synaptoneurosomes prepared from rat neocortex during a narrowly defined period of postnatal development. No glutamate stimulation was observed on the day of birth, even though high levels of phosphoinositide (PIns) turnover were observed with the muscarinic agonist carbachol. Glutamate-stimulated PIns turnover reached a maximum at one week of age, and decreased to adult levels by five weeks of age. Of the glutamate analogs tested, only ibotenate produced significant stimulation. N-Methyl-D-aspartate (NMDA) showed negligible stimulation and kainate showed only minor stimulation at the highest concentration tested (1 mM). Glutamate stimulation was not blocked by either the NMDA receptor antagonist 2-amino-5 phosphonovaleric acid (APV) or the broad spectrum glutamate antagonist kynurenic acid. These results suggest that a specific subclass of excitatory amino acid receptor linked to PIns metabolism, or a phospholipase associated with the receptor, is transiently expressed in the neocortex during early postnatal development. PMID- 2567642 TI - Biochemical diagnosis of Hunter syndrome on Epstein-Barr virus-transformed lymphoblastoid cell lines. AB - Long-term lymphoblastoid cell lines have been established from a patient with Hunter syndrome, from his mother, an obligate heterozygote, and from several control individuals. Biochemical analyses show that lymphoblastoid cells represent a suitable biological material for the diagnosis of hemizygous, affected males and for heterozygous females: clonal analysis demonstrates the mosaicism predicted by the Lyon hypothesis. PMID- 2567643 TI - Linkage in a family with X-linked Charcot-Marie-Tooth disease. AB - The gene for the X-linked form of Charcot-Marie-Tooth disease (CMT Peroneal Muscular Atrophy, X-linked: McKusick No. 30280) has been shown in a single family to be linked to DXYS1 with a lod score of 4.55 at a recombination fraction of 0.03 and to PGK1 with a lod score of 3.34 at zero recombination. This is in agreement with previous work based on several families. Pooled data from this family and 7 previously reported families give a maximum lod score of 12.04 at theta max of 0.05 for linkage between CMTX and DXYS1 loci. PMID- 2567644 TI - "Variability gene" effect of cholesteryl ester transfer protein (CETP) genes. AB - Cholesteryl ester transfer protein (CETP) may have important roles in transfer of lipids from cells to serum lipoproteins or between circulating lipoprotein particles. Restriction fragment length polymorphisms (RFLPs) in DNA at the CETP locus have been detected. In the present study we have used RFLPs detectable with the restriction enzyme TaqI to examine if CETP influences serum lipid variability (as opposed to absolute lipid levels). We have compared within-pair difference in serum lipid and apolipoprotein levels in monozygotic twin pairs of various genotypes in the B polymorphism at the CETP locus and uncovered significant differences between genotypes. We conclude that the CETP locus has "variability genes" (as opposed to "level genes") with respect to total and LDL cholesterol variability. A person's total genetic risk for coronary heart disease may depend on his or her combination of "level genes" and "variability genes". The method of analysis applied may be the best available for the study of gene - environment interaction. PMID- 2567647 TI - [Drugs used in the therapy of peptic ulcer with special reference to H2 receptor blockaders]. AB - Drugs used in the management of peptic ulcer are reviewed with special attention to the possibilities offered by H2 receptor antagonists. In this context, histaminergic transmission and distribution of H2 receptors are briefly reviewed. Among other substances, special interest has been raised by H+ and K+-dependent ATPase inhibitors; the most recently introduced derivatives of these substances which are as yet in the experimental stage, are also listed. PMID- 2567645 TI - Bacteroides-specific IgG and IgA subclass antibody-secreting cells isolated from chronically inflamed gingival tissues. AB - The emergence of cells that produce IgG and IgA subclass antibodies to Bacteroides gingivalis (Porphyromonas gingivalis) fimbriae and lipopolysaccharide (LPS) antigens was examined in mononuclear cells isolated from inflamed gingiva of different stages (slight, moderate or advanced) of adult periodontitis (AP). Antigen-specific IgM, IgG (including IgG1, IgG2, IgG3 and IgG4) and IgA (including IgA1 and IgA2) producing cells were enumerated by the ELISPOT assay and were compared with total Ig-producing cells of each isotype or subclass. In advanced AP, the B. gingivalis fimbriae-specific IgG- and IgA-secreting cells represented 5% of total IgG- or IgA-secreting cells, while those from the moderate stage comprised approximately 1% of these two isotypes. Cells producing antibody specific for B. gingivalis LPS were observed at frequencies of 0.1% and 0.4% for IgG and IgA cells, respectively in the advanced stage. When IgG subclasses were analysed in moderate AP, the anti-fimbriae subclass responses were largely IgG1 (60%), followed by IgG2 (20%), IgG3 (10%) and IgG4 (10%). Fimbriae-specific IgG subclass responses were elevated in the advanced stage of AP, and IgG4 (40%) and IgG1 (30%) were dominant, followed by IgG3 (20%) and IgG2 (10%). IgA1 cells predominated in both the moderate and advanced stages, however a relative increase in IgA2 cells occurred in advanced AP. Mononuclear cells isolated from gingiva of AP patients did not contain cells producing antibody to antigens such as Escherichia coli K235 LPS, cholera toxin or the hapten dinitrophenyl coupled to bovine serum albumin. These results show that local IgG and IgA subclass responses occur to a protein antigen of a major periodontal disease (PD)-associated pathogen, B. gingivalis, and the increase in IgG4 and IgA2 responses may be associated with host protection. PMID- 2567648 TI - Response to bronchodilators. AB - Bronchodilator response is usually tested to establish reversibility of airflow obstruction, to aid in diagnosis, and to help plan long-term bronchodilator therapy. Among the various tests to establish bronchodilator response, FEV1, FEF25-75% or FEF50%, and FVC are the most widely used. However, it may be difficult to interpret changes in pulmonary function in COPD patients because of day-to-day variability. A 15 per cent increase in FEV1 or FVC and a 30 per cent increase in isovolume FEF25-75% or FEF50% above baseline are acceptable criteria for bronchodilator response. beta-Adrenergic agonists, theophylline, and anticholinergic agents are available for testing bronchodilator response. Inhalation of two puffs of a beta-adrenergic agonist aerosol from an MDI is the most convenient and practical approach for testing in the laboratory. Bronchodilator response is presumed to be beneficial in the treatment of asthma. However, in the treatment of COPD a positive response may not be a reliable guide for establishing long-term benefit. PMID- 2567649 TI - Conspectus. Psychopharmacologic treatment of insomnia in adults and the elderly. PMID- 2567646 TI - Pharmacokinetics of the newer benzodiazepines. AB - The assay methods used to determine the concentrations of the newer benzodiazepines include electron-capture gas-liquid chromatography, high performance liquid chromatography with ultraviolet detection, gas chromatography mass spectrometry, radioassay and radioreceptor assay. The method used frequently is the highly sensitive and specific electron-capture gas-liquid chromatography. Other methods are associated with limitations. The triazolo- and imidazolebenzodiazepines differ structurally from the 'classical' benzodiazepines such as diazepam, and offer distinct differences in pharmacological activity and in time-course of effect. Alprazolam and triazolam, both 1,4 triazolobenzodiazepines, have high affinities for the benzodiazepine receptor as do midazolam and loprazolam, which are 1,4-imidazolebenzodiazepines. Absorption is characteristically rapid, with peak alprazolam and triazolam concentrations occurring within 1 hour after oral administration. Sublingual administration results in peak alprazolam and triazolam concentrations that are higher and occur earlier than with the oral route. The volume of distribution of alprazolam and triazolam is approximately 1L. Alprazolam is 70% bound to plasma proteins and the extent of binding is independent of concentration. Similarly, triazolam is approximately 85% bound to plasma proteins, variability in binding being explained by variations in alpha 1-acid glycoprotein concentration. The 1,4 triazolo ring prevents the oxidative metabolism of the classical benzodiazepines which results in formation of active metabolites with long elimination half lives. Alprazolam is extensively metabolised: 29 metabolites have been identified in the urine, and its major metabolite, alpha-hydroxyalprazolam, has pharmacological activity. alpha-Hydroxyalprazolam and 4-hydroxyalprazolam are detectable in plasma in amounts which account for less than 10% of the administered dose. Mean alprazolam elimination half-life in healthy adult subjects ranges from 9.5 to 12 hours; liver disease prolongs alprazolam elimination, but renal insufficiency does not. Triazolam also undergoes oxidation and subsequent glucuronidation. alpha-Hydroxytriazolam is the major metabolite, in addition to which 4-hydroxyalprazolam and alpha-4-hydroxytriazolam have been identified in plasma and urine. The elimination half-life of triazolam ranges between 1.8 and 5.9 hours, while that of the conjugated metabolites is short, approximately 3.8 hours. Accumulation of triazolam or its metabolites after multiple doses does not occur. Liver disease prolongs triazolam elimination from the body, but renal disease does not.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2567650 TI - The ratio of extracellular Ca2+ to K+ ions affects the photoresponses in Stentor coeruleus. AB - 1. Stentor coeruleus exhibits negative phototaxis (due to phototactic orientation response) and step-up photophobic response (avoiding reaction) to visible light. 2. The effect of Ja-value ([K+]/[Ca2+]1/2) and calcium ion concentration of the surrounding medium on the photoresponses in Stentor were studied. 3. The both types of photoresponses in Stentor are greatly affected by the Ja-value. A higher Ja-value medium suppressed the step-up photophobic response of Stentor, whereas the organism showed a higher degree of phototactic orientation response in higher Ja-value solutions. 4. The effect of the Ja-value on the step-up photophobic response was opposite to that on the phototactic orientation response. 5. With increasing calcium concentration but at a constant Ja-value, the number of Stentor showing the step-up photophobic response increased, whereas the phototactic orientation response of Stentor was suppressed at higher Ca2+ concentrations. 6. The effect of the calcium concentration on the photophobic response was also opposite to that on the phototactic orientation response, as in the case of Ja-value effect. PMID- 2567651 TI - The electroretinogram of the cockroach Leucophaea maderae. AB - 1. We have previously found that two components, the off-transient and the sustained component, of the electroretinogram (ERG) recorded from the eye of the cockroach Leucophaea maderae exhibit circadian rhythms in amplitude (Wills et al., 1985). 2. Analysis of the intensity-response functions of the two rhythmic components reveals that the circadian modulation of this visual system involves at least two independent regulatory processes. 3. Surgical and pharmacological manipulations suggest anatomically distinct sites of origin for the off-transient and the sustained component. 4. The results suggest that in this system the circadian pacemaker regulates visual processing both peripherally, at the level of the photoreceptors, and centrally, at the source of the off-transient. PMID- 2567652 TI - Cytotoxic effect of phenazine methosulphate on the isolated frog heart. AB - 1. Perfusion of isolated frog hearts with phenazine methosulphate (PMS) at 0.3 1.0 mM caused a fall in amplitude and frequency of beat, and finally a cessation of contractile activity, together with widespread ultrastructural damage. 2. Sarcolemma blebs were a characteristic feature of the damage. 3. No protection was provided by mannitol (10-100 mM), superoxide dismutase, catalase or a pHo of 6.6. 4. Potassium ferricyanide (1-6 mM), an artificial electron acceptor, also caused ultrastructural damage. 5. Comparisons are made with the oxygen paradox of mammalian heart, and the possible role of Ca2+ fluxes and oxygen radicals in muscle damage are discussed. PMID- 2567653 TI - A study of sulphate transport by lactating rat mammary tissue slices: evidence for anion exchange. AB - 1. The efflux of radiolabelled sulphate from lactating rat mammary tissue slices has been studied. Sulphate efflux was found to be time- and temperature dependent. 2. 4,4'-Diisothiocyanostilbene-2,2'-disulphonate (DIDS) inhibited a portion of sulphate release, whereas bumetanide was without effect. 3. The anions chloride, iodide and sulphate trans-stimulated sulphate efflux when added to the incubation medium. The increase in the efflux rate of sulphate found with chloride could be markedly inhibited by DIDS. Thiocyanate, unlike the other anions tested, only had a small effect. 4. The results strongly suggest that there is an anion exchange mechanism in the mammary gland which can mediate the transport of sulphate. This transporter may be important for the metabolism of sulphate by the mammary gland and may also help determine milk anion concentrations. PMID- 2567654 TI - Trypsin affects basal and stimulated osmotic water permeability in isolated toad skin. AB - 1. We investigated the effect of trypsin (Tryp) on basal, stimulated and fluphenazine (FPZ)-inhibited net water flow (Jw) through isolated toad skin (Bufo arenarum). 2. Epidermal Tryp (20 min) promoted an increase in basal Jw which was dose-dependent (maximal with 0.5 mg/ml) and was prevented by a Tryp inhibitor (SBTI). 3. Tryp treatment inhibited the subsequent response to substances known to act before (oxytocin, Oxy) or after cyclic AMP (cAMP) generation (theophylline). 4. Tryp-induced Jw was not additive with the maximal response to Oxy or theophylline and did not modify FPZ's inhibitory effect on stimulated Jw. 5. Dermal Tryp (0.5 mg/ml, 20 min) did not modify basal, but inhibited Oxy and isoproterenol-stimulated Jw, without altering the response to theophylline or db cAMP. 6. Collectively, our results show a differential action for epidermal and dermal Tryp. Tryp's side-selective action enables its use as a pharmacological tool in the functional dissection of Jw across toad skin. PMID- 2567655 TI - Non-shivering thermogenesis and obesity in adult diabetic Wistar fatty rats. AB - 1. Characteristics of resting and of norepinephrine (NE)-stimulated thermogenesis, and the glycemic response to NE were determined in adult male Wistar Fatty rats. Rats were maintained on Purina chow No. 5001 until 22 weeks of age, and fed semisynthetic diets containing 54% carbohydrate, 20% protein, 16% mixed fats, plus essential vitamins, minerals, and non-nutritive fiber from 22 until 30 weeks of age. 2. Obese rats were 50% heavier than lean throughout the study. Phenotype effects (obese greater than lean) were present for retroperitoneal (RP) and dorsal (DOR) white fat depot weight, adipocyte number per depot, and adipocyte lipid content. Epididymal mass and cellularity were similar in both phenotypes. 3. Interscapular brown adipose tissue (IBAT) mass, adipocyte size, and adipocyte number were greater in obese than in lean. Resting metabolic rates (RMR) of obese rats were lower than in lean, and increased 79% in lean but only 33% in obese animals following NE (200 micrograms/kg BW, s.c.) stimulation. 4. The glycemic response to NE occurred normally in both phenotypes, and resulted in a 3-fold increment in plasma glucose in lean rats and a 5-6-fold increase in plasma glucose in obese rats. 5. The results of this study are consistent with hyperplasia and hypertrophy of IBAT, RP and DOR depots, and indicate that the capacity for non-shivering thermogenesis is impaired in the obese phenotype of this strain in spite of peripheral sensitivity to NE and greater mass and cellularity of brown adipose tissue. PMID- 2567656 TI - Rapid clearance of Plasmodium yoelii-infected erythrocytes after exposure to the ionophore A23187. AB - 1. The effects of Ca2+ and the calcium ionophore A23187 on the intraerythrocytic development of the asexual forms of Plasmodium yoelii were examined. 2. Erythrocyte-free parasites obtained by saponin lysis of infected cells remained viable after exposure to 1 mM Ca2+. 3. A23187 inhibited the growth of P. yoelii and the inhibition was augmented by Ca2+ in cells infected with parasites at young stage of development. 4. A23187-treated infected cells disappeared from the circulation shortly after intravenous injection and this disappearance was profound in infected cells treated with the ionophore in the presence of Ca2+. PMID- 2567657 TI - Human granulocytes and granulocytes from other species demonstrate differences in chemotactic responsiveness to oxidized N-formyl-methionyl-leucyl-phenylalanine. AB - 1. Oxidation of the methionine of N-formyl-methionyl-leucyl-phenylalanine to the sulfoxide or sulfone derivative results in the loss of the peptide's chemotactic activity for human granulocytes. 2. The oxidized peptides are chemotactic for human monocytes; however, 10- to 100-fold higher concentrations are required for optimal monocyte chemotaxis. 3. Mouse, guinea pig and rabbit granulocytes, and the WBC264-9 human-mouse hybrid cell line migrated to the oxidized peptides and required 10- to 1000-fold higher concentrations of the oxidized peptides to elicit optimal chemotactic responses. 4. Human granulocytes appear to be unique in their lack of responsiveness to oxidized derivatives. PMID- 2567658 TI - Semantics as substance: life support systems or medically invasive technology? PMID- 2567659 TI - [Processes of pathologic coagulation and fibrinolysis in epidemic hemorrhagic fever with renal lesion]. AB - The processes of coagulation, anticoagulation and fibrinolysis as well as renal function were studied in 114 cases with epidemic hemorrhagic fever (EHF). The results showed that there were pathological coagulation and fibrinolysis and abnormal renal function in EHF. Comparing with normal controls, there was statistical significance (P less than 0.05) with the exception of fibrinogen. The degree of abnormality corresponded with the severity of the disease. With the observation of serial curves of these processes and renal function, we found out that there was correlation between them. Correlation analysis suggested that pathological coagulation and fibrinolysis could cause renal lesion or exacerbate it (P less than 0.05). The lesion affects mainly the glomerular filtration; whether it affects the tubular function or not is uncertain. PMID- 2567660 TI - [Study on the mechanism of glucose intolerance and insulin resistance in epidemic hemorrhagic fever]. AB - Intravenous glucose tolerance test (ivGTT) and insulin releasing test (IRT) were performed in 32 cases with epidemic hemorrhagic fever (EHF). 23 patients had impaired glucose tolerance, 17 patients showed hyperinsulinemia in fasting state, 12 patients had flat insulin response curves and most patients had higher plasma levels of growth hormone (GH), cortisol (F), norepinephrine (NE) and epinephrine (E) in critical stage than in convalescent stage. These data suggest that in EHF there are disturbed glucose metabolism and insulin resistance due to decreased pancreatic B cell reservoir and increased insulin antagonists such as GH, F, NE, E etc. It is suggested that use of insulin in addition to intravenous glucose infusion may be beneficial to glucose utilization in EHF. PMID- 2567662 TI - [ICO-10 monoclonal antibodies to the Thy-1 antigen]. AB - Mouse monoclonal antibodies (MAB) ICO-10 to Thy-1 antigen were obtained. MAB ICO 10 reacted in indirect immunofluorescence test with 5.7 +/- 0.8% human thymocytes. Antibodies did not react with granulocytes, monocytes, T- and non-T cells from peripheral blood, and with marrow cells of healthy donors. MAB ICO-10 reacted with blast cells from 25 of 53 patients with T-cell acute lymphoblastic leukemia (ALL), from 2 of 5 patients with B-cell ALL. This antigen was absent on blood and marrow cells from some patients with ALL, 80 patients with chronic lymphoid leukemia, 54 patients with chronic granulocytic leukemia at the stage of blastic crisis, 128 patients with acute nonlymphoblastic leukemia. Antibodies are specifically bound to thymocytes and spleen cells of Thy 1.1 and Thy 1.2 mice. MAB ICO-10 detect Thy-1 antigen expressed on human hematopoietic cells. MAB ICO 10 may be applied for human leukemia and lymphoma immune diagnosis. PMID- 2567663 TI - Sexual dimorphism and testosterone-dependent regulation of somatostatin gene expression in the periventricular nucleus of the rat brain. AB - Gender differences in hypothalamic somatostatin (SS) secretion may account in part for the sexually dimorphic patterns of GH secretion in rats. Since males have lower baseline serum GH levels than females, and SS inhibits GH secretion, we hypothesized that the SS neurons in the periventricular nucleus (PeN) of the male rat would have greater biosynthetic activity than those of the female. We tested this hypothesis by measuring SS mRNA in cells in the PeN of intact male and proestrous female rats. Using in situ hybridization and a computerized image analysis system, we measured SS mRNA content in individual cells in the PeN and compared signal levels (autoradiographic grains per cell) between male and proestrous female animals. The signal level of SS mRNA in cells of the PeN was significantly greater in males than in proestrous females (males, 210 +/- 7 grains/cell; females, 158 +/- 5 grains/cell; P less than 0.0005), whereas no difference was observed in SS cells of the frontal cortex (males, 100 +/- 0.8 grains/cell; females, 99 +/- 5.9 grains/cell). This difference in SS mRNA levels is likely to be the result of different hormonal environments exerting an influence on neurons of the hypothalamus. To test the hypothesis that testosterone stimulates SS gene expression in neurons of the PeN, adult male rats were castrated and immediately implanted with either empty (sham; n = 3) or testosterone-containing (n = 3) Silastic implants of a size that would deliver physiological levels of testosterone (3.6 +/- 1.5 ng/ml). We observed that castrated animals had significantly lower levels of SS mRNA signal in neurons of the PeN compared with intact animals (intact, 195 +/- 3 grains/cell; castrated, 159 +/- 6 grains/cell; P less than 0.003) and that physiological levels of testosterone prevent this reduction in SS mRNA levels (castrated testosterone replaced, 182 +/- 4 grains/cell; castrated, 159 +/- 6 grains/cell; P less than 0.003). Furthermore, testosterone-treated castrates had SS mRNA signal levels indistinguishable from those of intact controls (intact, 195 +/- 3 grains/cell; castrated testosterone-replaced, 182 +/- 4 grains/cell). There was no significant difference in SS mRNA levels in neurons of the frontal cortex (intact, 98 +/- 2 grains/cell; castrated, 98 +/- 3 grains/cell; castrated testosterone-replaced, 102 +/- 2 grains/cell).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2567664 TI - Mechanisms by which low salt condition increases sensitivity of thyroid stimulating antibody assay. AB - Mechanisms for high sensitivity in the assay of thyroid stimulating antibody under low salt (NaCl-deprived) hypotonic condition were analyzed using FRTL-5 rat thyroid cells. First, intracellular and extracellular cAMP contents after stimulations with TSH and Graves' immunoglobulin (Ig) were measured under both low salt (hypotonic) and high salt (isotonic) conditions. Higher total and much higher extracellular cAMP production were observed under the low salt condition. Under the high salt condition, the intracellular cAMP content reached a plateau after 10 min, whereas it increased progressively up to 120 min under the low salt condition. Thus, other cellular mechanisms, as well as increase in membrane permeability, seem to be involved in the enhanced response under the low salt condition. On the other hand, pretreatment of the cells with low salt solution without stimulator resulted in a decrease in subsequent cAMP response to TSH or Graves' Ig. Furthermore, [125I]bovine TSH binding to the cells under low salt condition also decreased after the cells were pretreated with the low salt solution. Scatchard analysis revealed that this decrease was due to a decrease in the Cmax of the low affinity binding site of the TSH receptor. Presence of stimulatory ligands to the TSH receptor from the beginning of exposure of the cells to low salt conditions was considered necessary. When the cells had been pretreated at 4 C with stimulator preparations in low salt condition, subsequent incubation at 37 C without stimulator resulted in more cAMP production than the case of high salt pretreatment. Moreover, low salt ligand-free incubation after the high salt binding resulted in higher cAMP production than high salt ligand free incubation after the low salt binding. Thus, both initial binding and incubation under the low salt condition appear capable of augmenting cAMP production. The fact that the sensitivity of the low salt assay to Graves' Ig was higher than that to bovine TSH was found to relate closely to the increase in initial binding. In conclusion, augmentation of cAMP production under low salt conditions is considered to be related not only to increased ligand binding and increased membrane permeability, but also to some alterations of postreceptor mechanisms during incubation. The proposed postreceptor mechanisms seem to be induced by stimulator binding, which prevents the TSH receptor from damage and promotes intracellular cAMP production progressively. Details of this remain to be elucidated. PMID- 2567665 TI - Amino acid availability and brain development: effects of nutritional and metabolic inadequacies. PMID- 2567666 TI - Continuation vs discontinuation of diet in phenylketonuria. PMID- 2567661 TI - Histamine H2-receptor antagonists versus prostaglandins in the treatment of peptic ulcer disease. PMID- 2567668 TI - Analysis of the biotin-binding site on acetyl-CoA carboxylase from rat. AB - The biotin-binding site of acetyl-CoA carboxylase from rat was characterized as to its amino acid sequence and relative position in the enzyme molecule. Biotin binds to the lysyl residue in the tetrapeptide Val-Met-Lys-Met; this tetrapeptide is located in close proximity to the NH2 terminus. In all other biotin-containing enzymes, the conserved tetrapeptide Ala-Met-Lys-Met is the counterpart to that of rat acetyl-CoA carboxylase; and the lysyl residue is 35 residues from the COOH terminus. To examine the significance of these unusual features of the biotinylation site of animal acetyl-CoA carboxylase, cDNA fragments were expressed in a bacterial system and the effects of specific site-directed mutagenesis were examined. Replacement of Val by Ala in the conserved tetrapeptide abolished biotinylation of the expressed protein. However, introduction of a termination codon at residue 36, in such a way that the distance between the lysine on which biotin binds and the COOH-terminal amino acid was 35 residues and the penultimate amino acid was the hydrophobic residue leucine, increased the efficiency of biotinylation, provided a substantial portion of the NH2-terminal peptide was removed. PMID- 2567667 TI - Ramp work tests with three different beta-blockers in normal human subjects. AB - The effects of beta-blockade on the responses of oxygen uptake (VO2), heart rate (HR) and blood lactate (La-) were examined during ramp cycle ergometer tests (50 W.min-1 ramp slope) in 8 healthy male volunteers. Each subject took placebo, or one of four different doses of three different beta-blockers (propranolol, metoprolol or oxprenolol) 2 h prior to each test for a total of 15 exercise tests. VO2 was measured breath-by-breath, HR was sampled once per breath, and La- was obtained every minute. Linear regression analysis was applied to VO2 and HR data to obtain the kinetic parameter total lag time (TLT) and a slope value. La- was analyzed by a continuous exponential model with the lactate slope index (LSI) being derived from the individual response curves. Submaximal exercise HR was significantly depressed at the baseline as well as during the ramp tests by beta blockade. TLT for HR was significantly affected by beta-blockade, with a dose dependent shift from a placebo value of 16 to 26 s with placebo to a value of -40 to -60 s at the highest dose. Slope of HR was significantly depressed relative to placebo. VO2 kinetics assessed by TLT were not significantly affected by beta blockade. This slope of the VO2 vs work rate relationship was significantly less than placebo only at the highest dose of beta-blocker. The LSI was not significantly affected by beta-blockade. In contrast with the clear impairment of HR response to exercise during beta-blockade, both the VO2 and La- responses appear to be relatively unaffected by beta-blockade during ramp exercise tests.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567669 TI - Insulin antagonism of dexamethasone induction of tyrosine aminotransferase in cultured fetal hepatocytes. A correlation between enzyme activity, synthesis, level of messenger RNA and transcription. AB - Previous studies have shown that insulin depresses the induction of tyrosine aminotransferase by glucocorticoids in cultured fetal rat hepatocytes. However, the site at which this inhibitory effect is exerted was not elucidated, since only enzyme activity was determined in such studies. Therefore, the effect of insulin on tyrosine aminotransferase synthesis, the level of its mRNA as well as the rate of transcription of the gene in isolated nuclei have been determined. The results obtained indicate that in cultures exposed to dexamethasone, Bt2cAMP, insulin and combinations of these additives, there is an excellent correlation between the enzyme activity, enzyme synthesis and the level of mRNA. Run-on transcription experiments indicate that the reduction in the level of mRNA by insulin in dexamethasone-supplemented cultures is the result of a diminished rate of gene transcription. PMID- 2567670 TI - Responses of glomerular filtration, renal blood flow and salt-water handling to acute cardioselective and non-selective beta-adrenoceptor blockade in essential hypertension. AB - The effects on renal hemodynamics and salt-water handling of equipotent doses of the cardioselective beta-blocker metoprolol (M, 100 mg) and of the non-selective (intrinsic sympathetic activity) beta-antagonist pindolol (P, 10 mg) were compared in 30 WHO Grade 1-2 hypertensive men. M lowered pulse rate more than P. Systolic pressure was equally reduced by both agents, and diastolic and mean pressures were decreased only after P. Glomerular filtration rate was not significantly altered by either antagonist, and renal blood flow decreased by approximately 11% both after M and P. Renal vascular resistance was unchanged after P, and was increased by 10% after M. It is concluded that, like the effects on central haemodynamics, ISA is more important in the renal response to beta adrenoceptor blockade than is beta-receptor selectivity. PMID- 2567671 TI - Modulation of catecholamine effects during hypoglycaemia in man by urapidil and propranolol. AB - The hypothesis that in hypoglycaemia adrenoceptor-blocking drugs may enhance those metabolic effects of the catecholamines that remain unblocked has been investigated in 12 volunteers. alpha-Adrenoceptor blockade with urapidil increased the heart rate and the plasma noradrenaline level, and increased the beta-adrenoceptor mediated cellular uptake of potassium and phosphate, and the production of lactate. Posthypoglycaemic glucose intolerance and the counterregulatory responses of hGH and cortisol remained unchanged. Plasma adrenaline, the alpha-adrenoceptor mediated responses of cortisol and hGH, and the diastolic blood pressure were increased by propranolol. Adrenoceptor blocking drugs produce an indirect stimulatory effect by eliciting a reflex increase in sympathetic tone, which is manifested as stimulation of receptors of the type that has not been blocked. PMID- 2567672 TI - A comparison of the extent and duration of hypokalaemia following three nebulized beta 2-adrenoceptor agonists. AB - The hypokalaemic effects of equal doses (5 mg) of fenoterol, salbutamol, terbutaline and an equal volume of saline administered by nebulization were compared in eight healthy subjects. Plasma potassium was measured at 15-min intervals for 60 min and at 90 min, 2, 4 and 6 h following administration. Fenoterol, salbutamol and terbutaline all significantly decreased plasma potassium when compared to saline; however, the magnitude and duration of this effect differed between the active agents. Both fenoterol and terbutaline significantly reduced plasma potassium for 4 h whereas salbutamol was only different from 30 to 120 min. The maximum decrease occurred with fenoterol (-0.78 mmol/l), followed by terbutaline (-0.70 mmol/l) and salbutamol (-0.33 mmol/l). Both terbutaline and fenoterol had a significantly greater effect compared with salbutamol. When administered by nebulization fenoterol and terbutaline are likely to have a greater hypokalaemic effect than salbutamol and this effect is likely to be more long lasting. PMID- 2567673 TI - Enhanced responsiveness of human memory T cells to CD2 and CD3 receptor-mediated activation. AB - Previous investigations have defined phenotypic differences between unprimed (naive) and antigen-primed (memory) T cells from human peripheral blood. We now report that memory T cells proliferate much more than naive cells when stimulated with anti-CD3 monoclonal antibody or pairs of anti-CD2 monoclonal antibodies. Enhanced responsiveness to receptor-mediated triggering is a novel mechanism for T cells which could facilitate memory cell response to specific antigen. Furthermore, when triggered via either CD2 or CD3, memory T cells produce substantial amounts of interferon gamma while naive cells produce virtually none; this suggests that differentiation from naive to memory state is accompanied by a stable change in regulation of the gene for interferon-gamma. We conclude that naive and memory T cells are dramatically different in function as well as phenotype. PMID- 2567674 TI - T cell activation via the CD2 molecule is associated with protein kinase C translocation from the cytosol to the plasma membrane. AB - T cell activation via the CD2 molecule involves phospholipase C and phosphoinositide hydrolysis. Here we demonstrate that the triggering of subclones of the human T leukemia Jurkat cell line by anti-CD2 as well as anti-CD3 monoclonal antibodies is able to induce activation (i.e. translocation from cytosol to cell membrane) of protein kinase C (PKC), which is dependent on the formation of 1,2-diacylglycerol from inositol 4-5-bisphosphate. The kinetics of PKC translocation parallels the rise in intracellular calcium following both CD2 and CD3 stimulations. These results further demonstrate that CD2 and CD3 activation pathways use similar signal transduction mechanisms. PMID- 2567675 TI - The influence of cyclosporin A on the alternative pathways of human T cell activation in vitro. AB - To gain further insight into the mechanism of action of the immunosuppressant cyclosporin A (CyA), we investigated the influence of CyA on proliferative responses of human T lymphocytes, induced via different membrane molecules. As was previously shown, activation of T cells via the T cell receptor (Ti)/CD3 complex with an anti-CD3 monoclonal antibody was inhibited by CyA. Likewise, triggering of T lymphocytes via the alternative, CD2(T11)-mediated pathway of activation was strongly inhibited. In contrast, responses induced by phorbol myristate 13-acetate (PMA; 100 ng/ml) or the combination of an anti-CD28 monoclonal antibody and a suboptimal concentration of PMA (1 ng/ml) were found to be insensitive to CyA. CyA-induced inhibition of both anti-CD3- and anti-CD2 mediated proliferation could not be reversed by addition of either PMA (1 ng/ml) or anti-CD28. An increase in the intracellular free Ca2+ concentration [( Ca2+]i) is an early event observed after stimulation of T cells via CD3 or CD2, whereas stimulation with PMA and anti-CD28 does not lead to a rise in [Ca2+]i. This suggests that the inhibitory action of CyA is related to Ca2+-dependent signaling pathways. Since we observed that CyA does not interfere with anti-CD3- or anti CD2-induced increases of [Ca2+]i, our data suggest that CyA-mediated inhibition is related to a later event in these intracellular signaling pathways. PMID- 2567676 TI - The effects of anti-CD2 antibodies on the differentiation of mouse thymocytes. AB - Using a rat monoclonal antibody against mouse CD2, we determined the expression of this marker on thymocytes during ontogeny. CD2 expression becomes detectable at day 15 and reaches adult levels (approximately 95% positivity) by day 19. Furthermore, the effect of anti-CD2 antibodies on T cell differentiation was analyzed by addition of antibodies to thymus organ cultures or repeated injection into newborn mice. Anti-CD2 antibodies inhibit CD2 expression in organ cultures and drastically reduce its expression on thymocytes and peripheral lymphocytes in vivo. In either situation, suppression of CD2 expression does not significantly alter the generation of T cells expressing CD3, CD4, CD8 and T cell receptor V beta 8. These results do not support a role for CD2 in early steps of thymocyte differentiation. PMID- 2567677 TI - Further studies on the involvement of dopamine and somatostatin in the inhibitory control of the growth and steroidogenic capacity of rat adrenal zona glomerulosa. AB - Prolonged (12-day) sodium restriction increased basal plasma concentration of aldosterone and provoked a notable hypertrophy of the zona glomerulosa and its cells in rats. A 7-day infusion of dopamine or somatostatin, at a rate which was found to exert a maximum inhibition of aldosterone secretion in 12 h, only partially reversed the effects of sodium deprivation. However, the combined administration of these two molecules not only completely annulled the effects of sodium restriction, but also lowered plasma aldosterone concentration and the volumes of the zona glomerulosa and its cells below the values found in rats fed a normal diet. These findings confirm the contention that dopamine and somatostatin are both involved in the negative control of the growth and steroidogenic capacity of the rat zona glomerulosa, and suggest that different mechanisms underlie the antiadrenoglomerulotrophic action of these molecules. PMID- 2567679 TI - Treatment compliance and self-medication in asthma in France. AB - A study was carried out among asthmatic patients in order to determine their compliance to treatment and self-medication. Data collection used a self administered postal questionnaire as tested in a previous study. The population under study included 450 asthmatic patients; 370 answered; their mean age was 47 +/- 16 yrs and 44% were men. An aerosol was the preferred route of administration (41%). Eighty two percent of the patients tended to reduce the doses following improvement; 80% claimed to know treatment for attacks; 90% had already been using an inhaler, although only 62% had been shown how to use one. Forty eight percent had already bought drugs without prescription, especially patients within the 25-34 yrs age group. This study is consistent with similar surveys performed in other populations. Improved quality of information about treatments and how to use drugs is higher in patients with chronic forms of asthma involving unpredictable acute attacks. PMID- 2567678 TI - Cytochalasin D inhibits L-glutamate-induced disc shedding without altering L glutamate-induced increase in adhesiveness. AB - Excitatory amino acid-stimulated disc shedding is correlated with the appearance of microfilament-rich ensheathing processes of the retinal pigment epithelium (RPE) and increased apparent adhesiveness between photoreceptors and RPE in explanted eyecups of Xenopus laevis. We have compared the time course of disc shedding and increased retinal adherence during L-glutamate treatment. Increased adherence was measured on the basis of the tendency of the apical RPE domains to partition with isolated neural retinas. In medium supplemented with L-glutamate (12 mM) or kainate (100 microM), a glutamate analog, the time course of increased partitioning of melanin pigment-rich cell fragments which contain ensheathing processes differs, even though the kinetics of induced disc shedding is the same in either case. Co-treatment with cytochalasin D (5 microM) completely blocks L glutamate-induced disc shedding, as well as formation of microfilament-rich ensheathing processes, even though it has little effect upon apparent adhesiveness. The virtually complete dissociation of the effects of L-glutamate on disc shedding from that on increased adhesiveness of photoreceptors to RPE suggests that increased retinal adherence and pseudopod formation may be unrelated causally. PMID- 2567680 TI - Muscarinic receptor modulation of glucose-induced electrical activity in mouse pancreatic B-cells. AB - Acetylcholine (1-10 microM) depolarized the membrane and stimulated glucose induced bursts of electrical activity in mouse pancreatic B-cells. The acetylcholine effects were mimicked by muscarine while nicotine had no effect on membrane potential. Pirenzepine, an antagonist of the classical M1-type muscarinic receptors, but not gallamine (1-100 microM), an antagonist of the classical M2-type receptors, antagonized the acetylcholine action on glucose induced electrical activity (IC50 = 0.25 microM). Bethanechol, an agonist of the classical M2-type muscarinic receptors, was approximately 100 times less effective than acetylcholine in stimulating the electrical activity. In addition, acetylcholine (1 microM) induced a marked increase (25%) in input resistance to the B-cell membrane. The results indicate that acetylcholine exerted its effects on the B-cell membrane by inhibiting K+ conductance via activation of a muscarinic receptor subtype distinct from the classical M2-type receptor. PMID- 2567681 TI - Interaction of cytosol fractions containing activated glucocorticoid-receptor complexes from rat liver and thymus with heterologous nuclei: effects on transcription. AB - Two rat liver cytosol fractions containing activated glucocorticoid-receptor complexes are able to stimulate the transcriptional activity of rat liver nuclei; the respective fractions from the cytosol of thymocytes inhibit the capacity of thymus nuclei for RNA synthesis. A similar inhibitory effect on thymus nuclei is exerted by the presence of rat liver cytosol fractions. Spot hybridization using a tyrosine aminotransferase (TAT) probe demonstrates that TAT gene expression is stimulated by the liver cytosol fractions acting on homologous nuclei whereas it is inhibited, in thymus nuclei, by the addition of thymus cytosol fractions. No effect on transcription is observed if the liver or thymus cytosol is heat activated in the presence of the glucocorticoid antagonist, cortexolone. Treatment of liver nuclei, previously subjected to the action of thymus cytosol fractions with the respective liver ones, restores transcriptional activity to control or higher levels. We conclude that rat thymocyte nuclei and cytosol contain transcriptional factors, which in the presence of the glucocorticoid receptor complex, irrespective of its source, inhibit gene expression, whereas in the absence of such factors, the glucocorticoid-receptor complex positively regulates the respective genes. PMID- 2567682 TI - Peptide hormone influence on exocrine pancreatic cancer. AB - At the present time there is only scant knowledge about the action of peptide hormones on pancreatic cancer--less than that of the sex hormones. However, the knowledge is rapidly expanding, and this research field contains many potentially very interesting effects and interactions. It seems possible that at least adjuvant effects of the peptide hormones can be expected to have a part to play in treatment of exocrine pancreatic cancer in the future. It is also of great theoretical interest to note that this 'target organ' for peptide hormones and 'non-target organ' for sex hormones may be influenced by peptide hormones as well as sex hormones. The presence of hormonal receptors and hormonal-binding proteins and their role in tumor genesis are not fully understood, but the discovery of cancer specific hormone-binding proteins in pancreatic cancer might be used in the detection and treatment of this cancer in the future. PMID- 2567683 TI - Genetic studies in inbred BB/Wor rats. Analysis of progeny produced by crossing lymphopenic diabetes-prone rats with nonlymphopenic diabetic rats. AB - BB/Wor diabetes-prone (DP) rats are lymphopenic and frequently develop insulin dependent diabetes. Diabetes-resistant (DR) BB/Wor rats are not lymphopenic and become diabetic rarely and at a significantly younger age. To examine the genetic basis for diabetes, lymphopenia, and age at onset of diabetes among inbred BB/Wor rats, we crossed nonlymphopenic diabetic rats with lymphopenic DP animals and studied F1, F2, and backcross progeny. F1 rats were neither diabetic nor lymphopenic. Diabetes (both types) and lymphopenia reappeared among F2 rats, confirming the permissive association of diabetes and lymphopenia and the recessive nature of both. The absence of diabetes in F1 rats also suggested that the combination of genes responsible for diabetes among lymphopenic and nonlymphopenic rats may be distinct. Nonlymphopenic parental, F1, and F2 rats revealed normal lymphocyte subsets, including CD8+ and RT6+ T-lymphocytes. Lymphopenic parental and F2 rats revealed the absence of CD8+ and RT6+ cells, indicating that these T-lymphocyte abnormalities of lymphopenic DP rats segregate with the lymphopenia gene. The distribution of the ages at onset of diabetes among F2 lymphopenic and F2 intercross rats was significantly earlier than among lymphopenic parental and backcross animals, suggesting that the age of diabetes onset is a heritable trait and that the gene(s) or genetic modifier(s) responsible for the earlier onset of F2 diabetes was acquired from the nonlymphopenic parents. Our genetic studies also confirmed the observations that the 2- and 7-kilobase Bam HI fragments of the MHC class I region do not correlate with diabetes or lymphopenia. PMID- 2567685 TI - Incidence of hemagglutination activity among pathogenic and non-pathogenic Bacteroides fragilis strains and role of capsule and pili in HA and adherence. AB - We analyzed the ability of 120 encapsulated strains of B. fragilis to agglutinate guinea pig and human red blood cells. Sixteen strains showed a strong hemagglutination (HA) ability, 21 strains a moderate HA ability, 7 strains a weak HA ability and 74 strains did not agglutinate the tested red blood cells. Six strains tested from each HA group were able to adhere to cheek epithelial cells and to a cultured human intestinal cell line. Hemagglutinating strains were the most adhesive. By electron microscopy, pilus-like structures were found in three of the encapsulated adhesive strains. Treatment of the bacterial cells with pronase E reduced both HA ability and adherence of piliated encapsulated, and of piliated non-encapsulated strains. Glucosidase treatment of cells reduced HA activity and adherence of piliated encapsulated and of non-piliated encapsulated strains. Finally, it was found that hemagglutinating strains are more frequently isolated from clinical specimens (55%) than from feces of healthy donors (20%). PMID- 2567684 TI - Indirect effect of catecholamines on development of insulin resistance in skeletal muscle from diabetic rats. AB - The role of an increased sympathetic activation in the development of insulin resistance in diabetic skeletal muscle was investigated. Epitrochlearis muscles from rats with streptozocin-induced diabetes and from controls were incubated in vitro for 0.5-12.0 h. Diabetes decreased maximal insulin-stimulated (20 mU/ml) glucose transport capacity by 60% (P less than .001), but this decreased insulin responsiveness returned to normal on in vitro incubation (3.79 +/- 0.59 before vs. 8.92 +/- 0.64 mumol.ml-1.h-1 after 12 h of incubation). The reversal of decreased insulin responsiveness in diabetic muscles did not require the presence of insulin and was not affected by the presence of 5.0 x 10(-8) M of epinephrine. However, it was possible to partially prevent the development of insulin resistance with regard to glucose transport by treating the rats with the beta adrenergic antagonist propranolol (0.5 mg/kg) every 12 h during the entire 72-h period in which the animals were kept diabetic (insulin responsiveness was 3.16 +/- 0.40 mumol.ml-1.h-1 for saline-injected group vs. 5.55 +/- 0.46 mumol.ml-1.h 1 for propranolol-treated group). This effect was not present after a single injection of the drug 2 h before the experiment or when propranolol treatment was withdrawn 12 h before the experiment. The beta-adrenergic blockade markedly reduced the plasma concentration of free fatty acids (0.5 +/- 0.01 mumol/ml for propranolol-treated rats vs. 1.1 +/- 0.1 mumol/ml for saline-treated rats; P less than .001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567686 TI - Cloning and expression of Thiobacillus versutus aspartate-semialdehyde dehydrogenase gene in Escherichia coli. AB - The Thiobacillus versutus asd gene coding for aspartate-semialdehyde dehydrogenase was cloned in Escherichia coli cells using pBR322 as a vector. The gene was expressed independently of its orientation, suggesting that E. coli RNA polymerase recognized T. versutus promoter sequence. The T. versutus DNA coded protein, of the molecular weight 44,000, was identified by the analysis of the proteins produced by minicells. PMID- 2567687 TI - Some strains of Escherichia coli of putative enteroadherent-aggregative serotypes produce an unusual fibrillar haemagglutinin. AB - Two strains of Escherichia coli that formed on unusual kind of mannose-resistant and eluting haemagglutinin (MREHA) reacting with the red blood cells of rat and mouse, when cultured at 37 degrees C but not at 18 degrees C, were examined by electron microscopy. Production of this rare rodent-positive MREHA was correlated with the presence of fine fibrillae of estimated diameter 2.5 nm that were demonstrated by negative staining and immuno-gold labelling with MREHA-specific anti-serum. These two strains belonged to serotypes 078:H- and 078:H33; thus, it would be useful to know whether enteroadherent-aggregative strains of E. coli of these and other serotypes also possess this unusual MREHA. PMID- 2567688 TI - [DNA fingerprinting of Rattus norvegicus: a new approach in genetic analysis]. AB - Recent finding in highly effective DNA probes for RFLP testing (of hypervariable minisatellite DNA type) has led to the invention of DNA fingerprinting--the new technique of great value for identification of individuals, establishing biological kinship and studies in population genetics. We anticipate that DNA fingerprinting procedure with M13 phage DNA as a probe which we have developed earlier, makes it possible to apply new approach in genetic analysis- establishing, whether or not a particular locus is associated with the inheritance of genetic disease, by comparing the whole restriction fragment data from affected and unaffected animals. In this work, using the method described we characterized the Kroushynsky-Molodkina rat strain with hereditary disposition for epileptic attacks and performed comparative fingerprint analysis of these defective and normal rat genomes. The data obtained may hold some promises for further seeking the particular defective gene. PMID- 2567689 TI - [Effects of flutropium bromide, a new antiasthma drug, with repeated administration on bronchomotor response and hepatic drug metabolizing enzymes]. AB - Effects of flutropium bromide, a new antiasthma drug possessing the quarternary ammonium structure of atropine derivatives, were investigated on the bronchodilatory activity with repeated inhalations in guinea pigs and on the hepatic drug metabolizing enzyme activities with repeated i.v. administration in rats. Single inhalation of flutropium bromide (0.03%) inhibited the ACh-induced bronchoconstriction without changing the fall in blood pressure induced by ACh. Flutropium bromide (0.03%) was inhaled for 5 min a day by placing the animal in an inhalation box, successively during periods of 14 and 28 days. The bronchodilatory effect of flutropium bromide after 14- or 28-day repeated inhalations was almost the same potency as that after single inhalation. Twenty eight-day repeated inhalations did not change the body weight curve in guinea pigs; and in addition, 14-day repeated i.v. administration did not change the hepatic drug metabolizing enzyme activities in rats. From the above results, it is indicated that flutropium bromide causes neither reduction in response nor cumulative effect after repeated inhalations, and that the agent maintains the bronchodilatory effect without change in the hepatic drug metabolizing enzyme activities. PMID- 2567690 TI - [Important progress for the ophthalmologist in basic genetic research]. AB - Ophthalmologists and human geneticists share a long standing interest in hereditary diseases and anomalies of the eye. Many of the primary genetic eye diseases are known, as ophthalmic symptoms are frequently part of a pleiotropic gene effect or the eye is affected secondarily. Progress in human genetics has also improved the understanding of genetic eye diseases. This can be demonstrated in the analysis of the function of color-vision genes and their abnormalities as well as the retinoblastoma gene. A line can be drawn from early formal analysis of pedigrees to cytogenetic mapping and, finally DNA analysis and sequencing of the involved genes. These advances have not only led to theoretical insights but also have practical applications where the determination of risk is concerned or prenatal diagnosis, genetic counselling, preventive measures and guidance. The retinoblastoma gene has become an important model for a tumor suppressor gene and tumorigenesis in general. Its influence on other types of tumors, such as osteosarcoma and breast cancer must be clarified. Sequencing of the gene opens the possibility of reconstructing the primary gene product by "reverse genetics" and of analyzing its mode of action. DNA analysis has been extended to an increasing number of eye diseases. Precise clinical and genetic analysis and diagnosis are of primary importance, however, for progress in this field. PMID- 2567692 TI - Benzodiazepines in alcoholism. PMID- 2567691 TI - Prescription of neuroleptics for geriatric nursing home patients. PMID- 2567693 TI - Genetic polymorphism of the apolipoprotein B gene locus influences serum LDL cholesterol level in familial hypercholesterolemia. AB - An XbaI restriction fragment length polymorphism (RFLP) within the coding region of the apolipoprotein B (apoB) gene has been found to be associated with serum cholesterol and triglyceride levels in several populations. Mutations in another genetic locus, the low density lipoprotein (LDL) receptor gene, give rise to familial hypercholesterolemia (FH), a disease characterized by hypercholesterolemia, tendon xanthomas and atherosclerosis. We determined the XbaI genotypes and serum lipoprotein levels of 120 unrelated patients with the heterozygous form of FH. A non-parametric analysis of variance showed a significant association between elevated serum total cholesterol concentration (P less than 0.05), serum LDL-cholesterol concentration (P less than 0.025) and the presence of the XbaI restriction site (X2 allele). Thus, patients homozygous for the presence of the XbaI restriction site (genotype X2X2, n = 28) had on average a 14% higher serum total cholesterol level and a 21% higher serum LDL-cholesterol level than those homozygous for the absence of this site (genotype X1X1, n = 29); patients heterozygous for the XbaI restriction site (genotype X1X2, n = 63) had intermediate serum total and LDL-cholesterol levels. No significant differences were seen in serum triglyceride or high-density lipoprotein (HDL)-cholesterol values between these patient groups. These data demonstrate that genetic polymorphism of the principal ligand for the LDL receptor, apoB, may contribute to serum cholesterol regulation, even in patients with grossly distorted cholesterol homeostasis. PMID- 2567695 TI - Mapping the mutation causing the X-linked lymphoproliferative syndrome in relation to restriction fragment length polymorphisms on Xq. AB - The X-linked lymphoproliferative syndrome (XLP) results in fatal infectious mononucleosis, hypogamma-globulinemia, and malignant lymphoma. The mutation has been mapped relative to several restriction fragment length polymorphism (RFLP) markers in the Xq21-Xq27 vicinity. The DXS37 locus was found to be near both the DXS42 and XLP loci. PMID- 2567694 TI - Maternal origin of a de novo chromosome 8 deletion in a patient with Langer Giedion syndrome. AB - The anonymous DNA probe L32, which defines the D8S48 locus within the Langer Giedion syndrome chromosome region on the long arm of chromosome 8, was used to search for a common restriction fragment length polymorphism. A HindIII and an MspI polymorphism were detected (polymorphism information contents 0.25 and 0.19, respectively). Both polymorphisms were informative in the family of a Langer Giedion patient carrying a de novo interstitial deletion 8q23-24.1. Lack of transmission of a maternal haplotype indicates that this deletion occurred during maternal gametogenesis. This finding contrasts with the frequent paternal origin of mutations in other microdeletion syndromes. PMID- 2567696 TI - Linkage analysis of seven kindreds with the X-linked lymphoproliferative syndrome (XLP) confirms that the XLP locus is near DXS42 and DXS37. AB - Analysis of seven kindreds indicates that the XLP locus exhibits 1% recombination with DXS42 (lod = 17.5) and no recombination with DXS37 (lod = 13.3). PMID- 2567697 TI - Human pepsinogen C (progastricsin) polymorphism: evidence for a single locus located at 6p21.1-pter. AB - A series of six clones containing the entire human pepsinogen C gene (PGC) was identified in a cosmid vector library by using cDNA and oligonucleotide probes. The 10.7-kb PGC gene includes nine exons and exhibits a high degree of sequence identity (60%) with the functionally related pepsinogen A genes. The predicted amino acid sequence was identical with the partial amino-terminal and carboxyl terminal sequences of purified pepsinogen C. An informative restriction fragment length polymorphism was detected with several restriction enzymes and involved an insertion or deletion of 100 bp of intron sequence located between exons 7 and 8. Evidence that there is only a single PGC gene in humans is presented. The PGC gene and the prolactin gene were regionally localized to 6p21.1-pter by analysis of mouse X human somatic cell hybrids. PMID- 2567698 TI - A chromosome 19 clone from a translocation breakpoint shows close linkage and linkage disequilibrium with myotonic dystrophy. AB - The gene for myotonic dystrophy (DM), the most common form of adult muscular dystrophy, is situated on the proximal long arm of chromosome 19. Although there exist markers that are tightly linked to the DM locus, its precise location is unknown. The identification and characterization of additional DNA probes closely linked to the DM locus continue to be priorities. In this study, we report on the linkage between a new DNA marker, designated p alpha 1.4P, and the DM locus in 50 families. The probe p alpha 1.4P was derived from a cloned breakpoint junction fragment from the chromosomal translocation t(14;19)(q32;q13.1). This translocation has been previously described in some cases of chronic lymphocytic leukemia. We have identified a BanI restriction fragment length polymorphism that is detected by p alpha 1.4P. Segregation analysis between this RFLP and DM revealed close linkage between the two loci (lod = 10.95, theta = 0). Furthermore, statistical evidence for linkage disequilibrium between p alpha 1.4P and the DM locus in a French Canadian population was found. Finally, by means of a somatic cell hybrid mapping panel, p alpha 1.4P was sublocalized to 19q12--- 19q13.2. PMID- 2567700 TI - Chromosomal localization of the human homeo box-containing genes, EN1 and EN2. AB - The human homologs of the mouse homeo box-containing genes, En-1 and En-2, which show homology to the Drosophila engrailed gene, have been isolated. The human EN1 gene was mapped to chromosome 2 by analysis of mouse-human somatic cell hybrids. The human EN2 gene was localized to chromosome 7, 7q32-7qter, by analysis of rodent-human somatic cell hybrids and cell lines carrying portions of chromosome 7. PMID- 2567701 TI - Mapping of Abll within a conserved linkage group on distal mouse chromosome 1 syntenic with human chromosome 1 using an interspecific cross. AB - A human Abelson related gene (ABLL) cDNA clone was used to detect restriction fragment length polymorphisms (RFLPs) on mouse Southern blots. Abll was mapped to mouse chromosome 1 by analysis of segregation with other distal chromosome 1 genetic polymorphisms by using a panel of DNAs from [(C3H/HeJ-gld/gld x Mus spretus) F1 x C3H/HeJ-gld/gld] interspecific backcross mice. The data indicate the following gene order: (centromere)-CD45-6.5 cM-Lamb-2-1 cM-Abll-2 cM-At-3. The results extend the analysis of a large conserved linkage group spanning nearly 30 cM on distal mouse chromosome 1 syntenic with human chromosome 1q21-32. Within this linkage group similar relative positions have been characterized in both species for C4BP, REN, CD45, LAMB2, ABLL, AT3, APOA2, and SPTA. PMID- 2567699 TI - Cloning of full-length methylmalonyl-CoA mutase from a cDNA library using the polymerase chain reaction. AB - The polymerase chain reaction was used to clone a full-length human methylmalonyl CoA mutase cDNA from a human liver library by priming with sequences from the 5' end of a partial cDNA and sequences in the phage vector. The amino acid sequence predicted from the cDNA corresponds to the authentic amino acid sequences of peptide fragment from purified methylmalonyl-CoA mutase. The open reading frame of the cDNA encodes 742 amino acids (82,283 Da) comprising a 32 amino acid mitochondrial leader sequence and a mature protein of 710 amino acids (78,489 Da). The use of the polymerase chain reaction to "screen" the cDNA library represents a novel application of this technique. The full length will enable analysis of mutations underlying inherited methylmalonic acidemias caused by deficiency of the methylmalonyl-CoA mutase apoenzyme. PMID- 2567702 TI - Detection and mapping of polymorphic KpnI alleles in the human T-cell receptor constant beta-2 locus. AB - Southern blot analysis with human T-cell receptor (TcR) beta-chain specific cDNA probes revealed two novel allelic forms of the TcR beta-2 gene locus. Three different genotypes were noted based on the presence of polymorphic KpnI restriction fragments: I, 5.7 kb fragment only; II, 3.9 kb and 1.8 kb fragments only; III, all three polymorphic fragments. This hybridization pattern suggested that the presence or absence of a polymorphic KpnI site within the 5.7 kb fragment defines the two different allelic forms of the TcR beta chain locus. By Southern blot analysis of genomic DNA from T-cell lines with deleted C-beta-1 regions and computer-assisted restriction site mapping of germline and cDNA sequences of the C-beta-2 locus, the polymorphic KpnI site was localized at 24 bp 5' to the third exon of the C-beta-2 gene. It was determined that the polymorphic KpnI site and the earlier described polymorphic BglII site located 5' to the C beta-2 gene are not co-inherited. No difference was noted in distribution of the KpnI genotypes and allelic frequencies between 26 normal individuals and 22 patients with systemic lupus erythematosus. However, this newly characterized polymorphism of the TcR locus should provide a useful tool to analyse the role of inherited genetic variations in the function of T lymphocytes under normal and pathological conditions. PMID- 2567704 TI - Giant gourami (Osphronemus goramy: Anabantoidei) as a potential agent for control of weeds, the breeding source for the vectors of Brugia malayi. AB - The biocontrol efficacy of giant gourami against various aquatic weeds, which form the breeding source of Mansonioides, the vectors of B. malayi, was explored. This fish showed a feeding predilection towards Pistia plant which is the most favoured host of immatures of Mansonioides. A single fish consumed this weed on an average of 206.25 +/- 19.09 g and 316.85 +/- 26.55 g a day under laboratory and field conditions respectively. Techniques developed for the culturing of this fish are also described. PMID- 2567703 TI - CD2 and other surface molecules in the regulation of non-MHC-restricted cytolytic function. AB - The effect of anti-CD2 and Fc receptor binding molecules on the cytolytic function of a highly enriched population of CD3- large granular lymphocytes (LGL) was studied. These cells could mediate natural killer (NK) activity, antibody dependent cellular cytotoxicity (ADCC) and lectin-dependent cellular cytotoxicity (LDCC). Both ADCC and LDCC were enhanced by anti-CD2. The enhanced LDCC could also be observed with IL-2-activated LGL. However, NK cell activity was usually slightly diminished or unaffected by anti-CD2 binding. Immune complex and aggregated human IgG had no effect on ADCC but an anti-CD16 showed a dose dependent inhibition of ADCC, reversible by anti-HLA-ABC and anti-CD2. Cross linking of LGL surface-bound anti-CD2 caused an almost complete inhibition of LDCC and ADCC but had much less effect on NK activity. These experiments show that ADCC and LDCC mediated by CD3- LGL can be influenced by perturbing the CD2 molecule. NK activity was, however, affected differently, suggesting some basic differences in the pathway of ADCC and NK function. PMID- 2567705 TI - Atrial natriuretic factor effects on cyclic nucleotides in a human renal cell line. AB - The natriuretic effects of atrial peptide hormones have been attributed, at least in part, to their stimulation of guanylate cyclase activity in renal cell membranes. The effects of atrial natriuretic factor (ANF) on stimulation of cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP) accumulation were investigated in cloned human kidney tumor (hKT) cells and parent cells from a human renal tumor epithelial cell line (SK-NEP-1). Human ANF (99-126) (10(-6)M) stimulated (p less than 0.001) cellular cGMP accumulation in a dose-dependent manner from a basal level of 0.26 +/- 0.04 to 3.73 +/- 0.81 pmol/mg protein/5 mi (mean +/- SEM, n = 13). ANF stimulation of cGMP accumulation was specific, in that high concentrations (10(-6)M) of atriopeptin I [rat ANF (103-123)], angiotensin II, arginine vasopressin, and amiloride (10(-4)M) did not increase basal cGMP. Amiloride (10(-4)M) enhanced (p less than 0.01, n = 6) the ANF stimulation of cGMP accumulation (1.24 +/- 0.39 pmol/mg protein/5 min), particularly at low doses of ANF (10(-10)M) where stimulation by ANF without amiloride (0.34 +/- 0.08 pmol/mg protein/5 min) was barely distinguishable from a basal level (0.19 +/- 0.02 pmol/mg protein/5 min) of cGMP accumulation. The stimulatory effect of ANF (1.59 +/- 0.07 pmol/mg protein/5 min) was attenuated (0.75 +/- 0.06 pmol/mg protein/5 min, p less than 0.01, n = 6) by preincubation of the cells with pertussis toxin but not by cholera toxin. ANF (4.56 +/- 0.93 pmol/mg protein/5 min, n = 8) did not affect cAMP accumulation (4.32 +/- 0.98 pmol/mg protein/5 min) in hKT cells. This is the first report of an ANF responsive human renal cell line, and its use should facilitate investigation of ANF-receptor interactions. PMID- 2567706 TI - Contractions to oxygen-derived free radicals are augmented in aorta of the spontaneously hypertensive rat. AB - To determine if oxygen-derived free radicals are mediators of endothelium dependent contractions to acetylcholine in the aorta of spontaneously hypertensive rats (SHR), the mechanism of contraction to xanthine plus xanthine oxidase was studied. Rings, with and without endothelium, of thoracic aorta from normotensive Wistar-Kyoto (WKY) rats and SHR were suspended in organ chambers for isometric tension recording. Oxygen-derived free radicals caused concentration dependent contractions; these contractions were twice as large in the aortas of SHR than in WKY rats. Deferoxamine reversed the response to xanthine oxidase to a small relaxation. Either allopurinol, superoxide dismutase, or catalase, or the combination of superoxide dismutase plus catalase reduced the contractions. Diltiazem inhibited the response to xanthine oxidase; in contrast, phentolamine plus propranolol did not affect it. Indomethacin and meclofenamate, but not tranylcypromine or dazoxiben blocked the contractions. Endothelium-dependent contractions to acetylcholine in aortas from the SHR were not affected by deferoxamine or superoxide dismutase plus catalase. These data suggest that hydroxyl radicals cause contractions in the rat aorta, which are dependent on extracellular calcium and mediated by activation of the cyclooxygenase in the vascular smooth muscle. The augmented contractions in the hypertensive strain are due to an increased reactivity of the smooth muscle to oxygen-derived free radicals. However, the lack of effect of the scavengers on endothelium-dependent contractions to acetylcholine suggests that the endothelium-derived contracting factor is chemically different from oxygen-derived free radicals. PMID- 2567707 TI - Aggravation of salt-induced hypertension in Dahl rats by 2% supplemental dietary calcium. AB - There is considerable interest in the antihypertensive potential of supplemental dietary calcium in salt-sensitive hypertension. Previously we reported that very high dietary calcium (4.0% vs. 0.4%) lowers mean arterial pressure in Dahl salt sensitive (DS) hypertensive rats. However, we have recently observed that more moderate calcium supplementation (2.0% vs. 0.4%) increases mean arterial pressure in DS rats. To further evaluate the pressor action of 2.0% versus 0.4% calcium, we tested for effects of 2.0% calcium in female DS rats fed low (0.2%), moderate (1.0%), and high (2.7%) sodium and in Dahl salt-resistant (DR) rats fed high sodium from 6 to 12 weeks old (n = 10-13 rats per group). At 12 weeks, 2.0% calcium increased mean arterial pressure and the cardiac ventricular weight/body weight ratio in DS rats fed high sodium (p less than 0.05) but not in DS rats fed low or moderate sodium or in DR rats fed high sodium. Ganglionic blockade decreased mean arterial pressure in all groups but failed to abolish or attenuate the difference in mean arterial pressure between high sodium-fed DS rats on 2.0% and 0.4% calcium diets. In the same DS rats fed a high sodium diet, 2.0% calcium increased systemic pressor responsiveness to graded norepinephrine administration after ganglionic blockade. Thus, 2.0% supplemental calcium intake enhances salt induced hypertension in DS rats. This prohypertensive action of 2.0% calcium is dependent on a critically high level of between 1.0% and 2.7% sodium in the diet.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567708 TI - Bradykinin antagonism and prostaglandins in blood pressure regulation. AB - These experiments were designed to analyze the interaction of a bradykinin antagonist with prostaglandins in blood pressure regulation of normotensive rats. Male Wistar rats, divided into three groups, received a 5-minute intra-arterial infusion of the bradykinin antagonist [( DArgo-Hyp3-Thi5,8-DPhe7]BK-TFA) at 250 micrograms/min. Groups were either intact rats (group I, n = 5), pretreated with indomethacin (group II, n = 10), or pretreated with both indomethacin and prazosin (group III, n = 8). The bradykinin antagonist infusion, which was shown to inhibit exogenous bradykinin by greater than 76% in intact animals, did not alter mean arterial pressure in group I rats despite a twofold increase in norepinephrine and a threefold increase in epinephrine. Group II rats presented a progressive increase in mean arterial pressure during the bradykinin antagonist infusion (14 +/- 3 mm Hg), with no statistically significant change in plasma catecholamines. Group III, with lower baseline mean arterial pressure due to alpha 1-adrenergic blockade, had an increase in mean arterial pressure comparable with group II during bradykinin antagonist infusion (22 +/- 5 mm Hg), confirming that this response was not sympathetically mediated. We conclude that in normotensive rats bradykinin plays a role in blood pressure regulation that is closely linked to that of prostaglandins and that points to a balance between these systems. PMID- 2567709 TI - Hemodynamic and renal responses to physiological levels of atrial natriuretic factor in conscious dogs. AB - The effects of increases in plasma atrial natriuretic factor (ANF) similar to those encountered after rapid volume expansion were examined in conscious dogs. Hemodynamics and renal function were continuously monitored during 30 minutes of human ANF infusion (10 ng/kg.min) and throughout a 30-minute recovery period. Ten minutes into the infusion period, plasma levels of ANF were elevated (p less than 0.01) by 34 +/- 9 from 36 +/- 5 pg/ml and sodium excretion increased (p less than 0.05) by 34 +/- 7 from 67 +/- 9 mueq/min. At that time, urine flow did not differ from baseline (0.25 +/- 0.03 ml/min). Renal blood flow velocity fell (p less than 0.01) by 5.0 +/- 0.5 from 42.3 +/- 3.7 cm/sec. Thirty minutes into the infusion period, plasma ANF levels were increased (p less than 0.01) by 61 +/- 9 pg/ml, similar to levels found after rapid volume expansion in conscious dogs. Urine flow and sodium excretion were elevated (p less than 0.01) by 0.35 +/- 0.06 ml/min and by 65 +/- 12 mueq/min, respectively. Renal blood flow velocity was reduced (p less than 0.05) by 4.4 +/- 1.5 cm/sec. Neither right atrial pressure, left ventricular end-diastolic pressure, mean arterial pressure, the first derivative of left ventricular pressure over time (dP/dt), nor heart rate were influenced by the elevated ANF plasma levels. Circulating levels of vasopressin and aldosterone were unaltered by these increases in plasma ANF. Thirty minutes into the recovery period, all variables were similar to the preinfusion baseline. Thus, in conscious dogs, physiologically relevant increases in plasma levels of ANF reached diuretic and natriuretic thresholds.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567710 TI - The effects of dopexamine hydrochloride on cardiopulmonary haemodynamics following cardiopulmonary bypass surgery. AB - We studied the effects of dopexamine hydrochloride in 14 patients following cardiopulmonary bypass for coronary revascularisation (12 patients) or valve replacement (2 patients). The drug was administered by intravenous infusion at rates of 1, 2, 4 and 6 micrograms/kilogram/minute for a period of 10 minutes at each dose. Measurements of heart rate, blood pressure, right atrial pressure, pulmonary arterial pressure, pulmonary arterial wedge pressure and cardiac output (thermodilution method) were made at the end of each period. There were significant increases in heart rate (P less than 0.05), cardiac index (P less than 0.05) and systolic blood pressure (P less than 0.05). There were significant falls in systemic vascular resistance (P less than 0.05) and pulmonary vascular resistance (P less than 0.05). There were no significant changes in pulmonary arterial or wedge pressure. No serious adverse effects were observed. Dopexamine hydrochloride appears to be a useful supportive agent in patients following cardiopulmonary bypass, although further trials need to be carried out in order to establish precise therapeutic indications for its use. PMID- 2567712 TI - Premature ovarian failure in a 35-year-old woman with a Robertsonian translocation. PMID- 2567711 TI - The ovary--then and now. PMID- 2567713 TI - Progesterone administration in patients with absent ovaries. AB - Forty-four women with absent ovaries who were referred to our centre for oocyte donation were treated with oestradiol valerate and natural progesterone. Serum concentrations and endometrial changes were evaluated on day 21 of stimulated luteal phases after the daily administration of 100 mg of natural progesterone in oil injected intramuscularly or after the ingestion of 300 mg of micronized progesterone or after vaginal administration of 300 mg or 600 mg micronized progesterone, respectively. Endometrial tissue did not show evidence of an adequate secretory pattern after orally administered progesterone. However, after intramuscular injection of 100 mg of natural progesterone and after the vaginal administration of 300 to 600 mg of micronized progesterone, the histologic and electron microscopic pictures were similar in all treatment groups and were close to the expected day 21 of the cycle. Serum levels of progesterone after intramuscular injection were five times higher than after vaginal administration. PMID- 2567714 TI - Daily or alternate-day FSH therapy in patients with polycystic ovarian disease resistant to clomiphene citrate treatment. AB - The effectiveness of intramuscular follicle stimulating hormone (FSH) administered as daily or alternate-day injections to patients with polycystic ovarian disease (PCOD) who previously failed to ovulate on clomiphene citrate was compared. The study comprised 20 treatment cycles of daily FSH and 19 of alternate-day FSH in 12 patients. The overall ovulation rate per cycle was 78% and was similar for both groups. Nine out of 12 patients achieved a pregnancy, with twice as many occurring in the alternate-day group. It is concluded that alternate-day FSH therapy is as effective as daily FSH in achieving ovulation and pregnancies in patients with PCOD resistant to treatment with clomiphene citrate. PMID- 2567715 TI - A controlled trial of intrauterine insemination for cervical factor and male factor: a preliminary report. AB - In a prospective, randomized, controlled trial, we evaluated the efficacy of intrauterine insemination (IUI) in 54 couples with cervical factor (CF) infertility and in 19 couples with male factor (MF) infertility. The diagnoses were confirmed by strict criteria and all other etiologies of infertility were ruled out prior to entry. Patients served as their own controls with alternating cycles of washed sperm IUI and whole-ejaculate intracervical insemination (ICI). In 113 paired cycles (mean 2.1 pairs/patient; range 1-7 paired cycles), 13% (n = 7) of CF patients became pregnant by IUI, 7% (n = 4) by ICI, and 6% (n = 3) by intercourse after missing inseminations. Seventy-one percent of IUI pregnancies occurred in the first 2 months. In 45 paired cycles for MF patients (mean 2.4 pairs/patient; range 1-6 paired cycles), two patients conceived, both in the first IUI cycle. These data suggest that pregnancies resulting from IUI occur during early treatment cycles. PMID- 2567716 TI - Progesterone production from granulosa cells of individual human follicles derived from diabetic and nondiabetic subjects. AB - Insulin and insulin-like growth factors have been implicated in the stimulation of ovarian steroidogenesis. To assess the effect of diabetes mellitus on this process, a comparison was made of progesterone production by cultured granulosa cells (50,000 cells/well) from 11 individual follicles of nondiabetic and 6 individual follicles of diabetic women. Diabetic metabolic control was fair [HbA1C 6.8, 8.7 (nl 5.0-7.5)]. Cells were collected by laparoscopic follicular aspiration after ovulation induction and isolated by Percoll gradient centrifugation. Progesterone production was measured after culture with hCG (10 IU/mL) or insulin (100 microU/mL). In both nondiabetic and diabetic groups on day 4, hCG significantly stimulated progesterone production (1,686 +/- 1,268 ng/mL to 4,123 +/- 2,825 ng/mL and 1,059 +/- 249 ng/mL to 1,506 +/- 245 ng/mL, respectively). In nondiabetic follicles, insulin also stimulated progesterone production on days 4 (2366 +/- 1032 ng/mL to 3699 +/- 1582 ng/mL; P less than .05) and 7 (987 +/- 475 ng/mL to 1858 +/- 929 ng/mL; P less than .05); this response was not noted in diabetic granulosa cells. We suggest that insulin stimulated progesterone production by granulosa cells isolated in the presence of fair diabetic metabolic control is impaired. PMID- 2567717 TI - Bromocriptine versus progesterone therapy for infertility related to luteal phase defects in hyperprolactinemic patients. AB - Several anecdotal reports suggested an association of luteal phase defects (LPD) and hyperprolactinemia. Some physicians treat LPD with ovulation-inducing drugs, whereas others recommend progesterone support of the luteal phase. A study was thus initiated to evaluate in cases of LPD associated with hyperprolactinemia which therapy would be more efficacious--bromocriptine or progesterone (P). LPD was divided into two types based on follicle dynamic studies: (1) LPD associated with immature follicles and (2) pure LPD when the follicle was mature. The objective was to determine if P or bromocriptine would be more effective depending on the type of LPD. Randomized therapy with either bromocriptine (BCT) or progesterone vaginal suppositories (PVS) was given to 60 patients with pure LPD (established by endometrial biopsy in the late luteal phase) and similarly randomized therapy was given to 40 women with LPD and immature follicles. The incidence of pregnancies during an 8-month treatment period was as follows: pure LPD--23 of 50 women (77%) treated by PVS versus 5 of 30 women (17%) treated by BCT; LPD associated with immature follicles--3 of 20 women (15%) treated by PVS versus 14 of 20 women (70%) treated by BCT. Those women failing to conceive were now given the alternate therapy for the next 8 months.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567718 TI - Immunoreactive relaxin in seminal plasma of man. AB - Relaxin in seminal fluid was determined radioimmunologically in 238 andrological patients with various ejaculate qualities. In 52 cases, a second ejaculate was examined 4 to 6 weeks later. Relaxin was found present in each ejaculate sample. There was no significant difference between the relaxin values of the reexamined cases and the original samples, indicating little fluctuation. Statistical correlations between relaxin levels in seminal plasma and clinical data, ejaculate, and hormonal parameters could not be established. Only cases with low fructose values showed some correlation with low relaxin levels in seminal plasma. PMID- 2567719 TI - Pulse labeling of hydroxyproline and leucine in reproductive tissues: a model for assessment of degree of reactivity. AB - A prospective study evaluating tissue reactivity by light microscopic and pulse labeling-autoradiographic techniques is presented. One uterine horn of a Sprague Dawley rat model was subjected to transection and reanastomosis using microsurgical techniques (control). The opposite horn of each was also cross sectioned with the application of an absorbable topical hemostatic agent (Avitene) and then reanastomosed. After closure of the abdomen, the animals were randomly assigned to groups according to the planned day of sacrifice (days 3, 7, 14, 21, or 28). Autoradiographic techniques utilizing [3H]hydroxyproline or [3H]leucine as well as light microscopy were employed to determine tissue reactivity. The results demonstrated an increase in stromal (P less than or equal to .05) and smooth muscle (P less than or equal to .01) reactivity with [3H]leucine labeling, and an increase in smooth muscle (P less than or equal to 0.05) reactivity with [3H]hydroxyproline in the Avitene-treated groups. It is concluded that pulse labeling with tritiated leucine and hydroxyproline is an effective method of objective analysis of tissue reactivity in reproductive organs. PMID- 2567720 TI - The influence of seminal plasma and polyaminic substances on the motility of isolated human sperm. AB - The purpose of the study was to examine the influence on the motility patterns of isolated human sperm of seminal plasma originating from semen of high and low motility, as well as of seminal plasma of azoospermic origin. Also assessed was the effect of incubation of sperm in the presence of solutions of the polyamines spermine, spermidine, and putrescine. The mean values of the obtained motility percentage and grade were related to the quality of sperm motility in semen and the quality of seminal plasma in that respect. The best results were obtained with specimens containing greater than or equal to 50% motile sperm and having a motility grade greater than or equal to 2. No relationship was found between the concentrations of polyamines in the seminal plasma and sperm motility of respective semen, as well as between the levels of polyamines and their effect on isolated sperm motility. It appears that the participation of polyaminic substances in the process of motility could be contributive and dependent on additional factors present in the semen. PMID- 2567721 TI - Acrosin inhibitor in extract of the zonae pellucidae of porcine oocytes. AB - Zonae pellucidae isolated from porcine oocytes were solubilized in 0.15 M NaCl at pH 2.5. The zonal extract contained a protein with relatively low molecular weight, which inhibited the amidase activity of boar acrosin and bovine trypsin. This protein was immunologically related to the BUSI II proteinase inhibitor isolated from bull seminal plasma. PMID- 2567723 TI - [Prevention of myocardial infarct with nitrates, beta blockers and calcium antagonists]. PMID- 2567722 TI - Effect of TSAA-291 on male reproductive tract and fertility of the rat. AB - Adult male rats were injected with TSAA-291 (16 beta-ethyl-17 beta-hydroxyestr-4 en-3-one), a steroidal antiandrogen, daily for 30 days at two doses, viz., 10.0 and 25.0 mg/kg. The treatment caused a dose-dependent reduction in the weights of testis, epididymis, and other accessory sex glands. In the animals that received the high dose, 50% of the rats showed spermatogenic arrest in about 20% of the seminiferous tubules and Leydig cell morphology was normal. The levels of glycerylphosphorylcholine and sialic acid were significantly reduced in the cauda epididymis of all the treated rats. There was a reduction in the number of motile spermatozoa in the cauda epididymis of rats treated with high dose of the antiandrogen. This was accompanied by a reduced number of females exhibiting spermatozoa in their vaginal smears. A combination treatment of the antiandrogen and androgen appears to be a promising approach for fertility regulation in the male. PMID- 2567724 TI - Laron-type dwarfism is associated with normal growth hormone and insulin-like growth factor I gene restriction patterns. PMID- 2567725 TI - The DNA polymorphisms of the beta-globin gene cluster and the arrangements of the alpha- and the gamma-globin genes in Koreans. AB - The DNA polymorphisms at the seven restriction sites in the beta-globin gene cluster in healthy Koreans were examined using four restriction endonucleases, Hinc II, Hind III, Ava II, and Bam HI. Seven (f = 0.326) and four individuals (f = 0.246) were homozygous for [+-----+] and [+----+-], respectively, among 66 individuals examined. As to the subhaplotypes 5' to the delta-globin gene, 25 (f = 0.615) and three individuals (f = 0.213) were homozygous for [+----] and [-+ ++], respectively. The frequency of [-++-+], which carries the A gamma T-globin gene, may be low in this population. It was recognized again that [+/+] at the Hinc II site 5' to the epsilon-globin gene always accompanied [--+--] at the Hinc II sites in and 3' to the psi beta 1-globin gene. The Korean, Japanese, and Chinese populations were not significantly different from each other in their haplotypes (subhaplotypes). The frequencies of the abnormal alpha- and gamma globin gene arrangements in Koreans were low. alpha-Thalassemia may occur at low frequency in Koreans. PMID- 2567726 TI - An allelic cluster of DQ alpha restriction fragments is associated with multiple sclerosis: evidence that a second haplotype may influence disease susceptibility. AB - Extensive analysis of restriction fragment length polymorphism using HLA class II and T-cell receptor gene probes has been carried out in an attempt to identify genetic markers more strongly associated with multiple sclerosis than the classically defined antigens DR2, Dw2, and DQw1. The use of DNA pooled from groups of patients and controls from northeast Scotland enabled the screening of 14 restriction endonucleases with five HLA-D region probes (DP alpha, DP beta, DQ alpha, DQ beta, DR beta) and two T-cell antigen receptor probes. Restriction fragment length polymorphisms which discriminated between multiple sclerosis and control pools were identified with four restriction enzymes: Msp1 (DQ alpha), BamH1, Bgl11, and Taq1 (DQ beta). No discriminatory polymorphism was seen with any of the other enzyme/probe combinations. Subsequent Southern blot analysis of individual DNA samples was carried out using these enzymes and probes in two independently conducted studies, in Northern Ireland and northeast Scotland. Following Msp1-digestion and hybridization to DQ alpha, a 3.25-kb fragment was observed in 31% of Scottish patients but in only 4% of controls from the same population. Furthermore, when only DR2-positive individuals were analyzed, there was a significant excess of this fragment in patients from both Scotland (28, or 2.9%) and Northern Ireland (20, or 3.4%). Although the DQ alpha gene characterized by this fragment remains to be determined, this fragment exhibits apparent allelism to DQw1. Therefore, these data raise the possibility that two different DQ alleles, one on each haplotype, may jointly contribute to disease susceptibility. PMID- 2567727 TI - Pseudoarylsulfatase A deficiency in a psychiatrically disturbed adolescent. AB - A case of pseudoarylsulfatase A deficiency in an adolescent boy presenting with affective lability, impulsivity, aggression, inattention, and academic difficulties is described. Genetically related to metachromatic leukodystrophy, pseudoarylsulfatase A deficiency has generally been felt to be a benign disorder. Pseudodeficiency of arylsulfatase A has, however, been associated with serious psychiatric morbidity in recent studies. Possible explanations for this association are suggested. To the best of the authors' knowledge, this is the first case report of pseudoarylsulfatase A deficiency in a psychiatrically disturbed adolescent. PMID- 2567728 TI - Characterization of amino acid transport in membrane vesicles from the thermophilic fermentative bacterium Clostridium fervidus. AB - Amino acid transport was studied in membrane vesicles of the thermophilic anaerobic bacterium Clostridium fervidus. Neutral, acidic, and basic as well as aromatic amino acids were transported at 40 degrees C upon the imposition of an artificial membrane potential (delta psi) and a chemical gradient of sodium ions (delta microNa+). The presence of sodium ions was essential for the uptake of amino acids, and imposition of a chemical gradient of sodium ions alone was sufficient to drive amino acid uptake, indicating that amino acids are symported with sodium ions instead of with protons. Lithium ions, but no other cations tested, could replace sodium ions in serine transport. The transient character of artificial membrane potentials, especially at higher temperatures, severely limits their applicability for more detailed studies of a specific transport system. To obtain a constant proton motive force, the thermostable and thermoactive primary proton pump cytochrome c oxidase from Bacillus stearothermophilus was incorporated into membrane vesicles of C. fervidus. Serine transport could be driven by a membrane potential generated by the proton pump. Interconversion of the pH gradient into a sodium gradient by the ionophore monensin stimulated serine uptake. The serine carrier had a high affinity for serine (Kt = 10 microM) and a low affinity for sodium ions (apparent Kt = 2.5 mM). The mechanistic Na+-serine stoichiometry was determined to be 1:1 from the steady-state levels of the proton motive force, sodium gradient, and serine uptake. A 1:1 stoichiometry was also found for Na+-glutamate transport, and uptake of glutamate appeared to be an electroneutral process. PMID- 2567729 TI - Isolation of nit-4, the minor nitrogen regulatory gene which mediates nitrate induction in Neurospora crassa. AB - Expression of nitrate reductase in Neurospora crassa requires the positive action of nit-4, a pathway-specific regulatory gene, which mediates nitrate induction. We report the molecular cloning of the nit-4 gene and present results which suggest that the nit-4 gene is constitutively expressed to yield a low-abundance 2.2-kilobase transcript. These results indicate that the nit-2 major control gene and the nit-4 pathway-specific control gene independently regulate the expression of the nitrate assimilatory structural genes. PMID- 2567730 TI - Stimulants in the treatment of depression: a critical overview. AB - The authors reviewed the efficacy and safety of stimulant drugs in the treatment of depression. Although uncontrolled studies were generally positive, the 10 placebo-controlled studies of stimulant drugs in primary depression, with one exception, indicated little advantage of drug over placebo. Although several of these studies were methodologically unsophisticated, they were comparable with and performed during the same period as studies establishing the efficacy of imipramine. Controlled studies of stimulants in apathetic or depressed geriatric patients were more likely to be positive, but outcome frequently consisted of partial improvement. Studies in medically ill patients with depression were promising but uncontrolled. Side effects have not been severe, and these drugs may pose less of a risk than tricyclics in the medically ill or elderly. Habituation is suggested, but there are no placebo-controlled studies to confirm this. In short, the stimulant drugs do not appear to be as effective as the conventional antidepressants in primary depression, but may be of value in refractory cases or in special cases, such as those involving the medically ill patient. Placebo-controlled trials are needed to explore these questions. PMID- 2567731 TI - Sterol-independent regulation of 3-hydroxy-3-methylglutaryl-CoA reductase by mevalonate in Chinese hamster ovary cells. Magnitude and specificity. AB - In this paper, we assess the relative degree of regulation of the rate-limiting enzyme of isoprenoid biosynthesis, 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase, by sterol and nonsterol products of mevalonate by utilizing cultured Chinese hamster ovary cells blocked in sterol synthesis. We also examine the two other enzymes of mevalonate biosynthesis, acetoacetyl-CoA thiolase and HMG-CoA synthase, for regulation by mevalonate supplements. These studies indicate that in proliferating fibroblasts, treatment with mevalonic acid can produce a suppression of HMG-CoA reductase activity similar to magnitude to that caused by oxygenated sterols. In contrast, HMG-CoA synthase and acetoacetyl-CoA thiolase are only weakly regulated by mevalonate when compared with 25 hydroxycholesterol. Furthermore, neither HMG-CoA synthase nor acetoacetyl-CoA thiolase exhibits the multivalent control response by sterol and mevalonate supplements in the absence of endogenous mevalonate synthesis which is characteristic of nonsterol regulation of HMG-CoA reductase. These observations suggest that nonsterol regulation of HMG-CoA reductase is specific to that enzyme in contrast to the pleiotropic regulation of enzymes of sterol biosynthesis observed with oxygenated sterols. In Chinese hamster ovary cells supplemented with mevalonate at concentrations that are inhibitory to reductase activity, at least 80% of the inhibition appears to be mediated by nonsterol products of mevalonate. In addition, feed-back regulation of HMG-CoA reductase by endogenously synthesized nonsterol isoprenoids in the absence of exogenous sterol or mevalonate supplements also produces a 70% inhibition of the enzyme activity. PMID- 2567732 TI - A novel bovine spinal cord endoprotease with high specificity for dynorphin B. AB - An endopeptidase that converts the opioid peptide dynorphin B (Tyr-Gly-Gly-Phe Leu-Arg-aRg-Gln-Phe-Lys-Val-Val-Thr) to its bioactive fragment Leu-enkephalin Arg6 was isolated from bovine spinal cord. The enzyme was purified about 230-fold from a concentrated spinal cord extract. Upon sodium dodecyl sulfate polyacrylamide gel electrophoresis, it stained as a protein of Mr 55,000. The purified enzyme is optimally active at around pH7 and has essential thiol groups. It appears to be highly specific for dynorphin B (Km = 11 microM) but not for alpha-neoendorphin or dynorphin A, two other opioids included in the prodynorphin precursor. From its specificity, molecular size, and inhibitory spectrum, this enzyme is different from other known dynorphin-converting or -degrading enzymes and appears to be a unique and novel endoprotease. PMID- 2567733 TI - Functional consequences of glutamate, aspartate, glutamine, and asparagine mutations in the stalk sector of the Ca2+-ATPase of sarcoplasmic reticulum. AB - Nucleotides encoding glutamate, glutamine, aspartate, or asparagine residues within the stalk sector of the sarcoplasmic reticulum Ca2+-ATPase were altered by oligonucleotide-directed site-specific mutagenesis. The mutant cDNAs were expressed in COS-1 cells, and mutant Ca2+-ATPases were assayed for Ca2+ transport function and phosphoenzyme formation. Multiple mutations introduced into stalks, 1, 2, and 3 resulted in partial loss of Ca2+ transport function. In most cases, subsequent mutation of individual amino acids in the cluster had no effect on Ca2+ transport activity. In one cluster, however, it was possible to assign the reduction in Ca2+ transport activity to alterations of Asn111 and Asn114. The mutant Asn114 to alanine retained about 50% activity, whereas the change Asn111 to alanine retained only 10% activity. None of the mutations affected phosphorylation of the enzyme by ATP in the presence of Ca2+ or by inorganic phosphate in the absence of Ca2+. The combined experiments suggest that the reduced Ca2+ uptake observed in the Asn111 and Asn114 mutants was not due to a defect in enzyme activation by Ca2+ or in formation of the phosphorylated enzyme intermediate but rather to incompetent handling of the bound Ca2+ following ATP utilization. These results demonstrate that the acidic and amidated residues within the stalk region do not constitute the high affinity Ca2+-binding sites whose occupancy is required for enzyme activation. They may, however, act to sequester cytoplasmic Ca2+ and to channel it to domains that are involved in enzyme activation and cation translocation. Simultaneous mutation of 4 glutamate residues to alanine in the lumenal loop between transmembrane sequences M1 and M2 did not affect Ca2+ transport activity, indicating that acidic residues in this lumenal loop do not play an essential role in Ca2+ transport. Similarly, mutation of Glu192 and Asp196 in the beta-strand domain between stalk helices 2 and 3 did not affect Ca2+ transport activity, although mutation of Asp196 did diminish expression of the protein. PMID- 2567734 TI - Glutamate overcomes the salt inhibition of DNA polymerase III holoenzyme. AB - Even though Escherichia coli can grow in media containing up to 1 M NaCl, one fifth that amount of NaCl will completely inhibit the in vitro activity of DNA polymerase III holoenzyme. It has been established that the major intracellular ionic osmolytes are potassium and glutamate (Richey, B., Cayley, D. S., Mossing, M. C., Kolka, C., Anderson, C. F., Farrar, T. C., and Record, M. T., Jr. (1987) J. Biol. Chem. 262, 7157-7164). We have found that holoenzyme catalyzes replication efficiently in vitro in up to 1 M potassium glutamate. Two salt effects on the replication of single-stranded DNA were observed. At low salt replicative activity was enhanced and at high salt there was anion-specific inhibition. We have found that DNA polymerase III holoenzyme tolerated 10-fold higher concentrations of glutamate than chloride. The ability of various anions to extend the useful range of salt concentrations followed the order: phosphate less than chloride less than N-Ac-glutamate less than acetate less than glycine less than aspartate less than glutamate. With the exception of phosphate, this order followed the Hofmeister series indicating that the anion-specific effects were due to anions interacting at the protein-water interface at weak anion binding sites. Glutamate did not reverse the inhibition by chloride. The low salt enhancement and high salt inhibition effects were additive for the two anions indicating that they competed for common anion binding sites. The major salt sensitive step was holoenzyme binding to template rather than the subsequent elongation reaction. PMID- 2567735 TI - Activation of glutamate by gamma-glutamate kinase: formation of gamma-cis cycloglutamyl phosphate, an analog of gamma-glutamyl phosphate. AB - gamma-Glutamate kinase, the enzyme that catalyzes the first step in the pathway from glutamate to proline, has been postulated to convert glutamate to a gamma activated form (possibly gamma-glutamyl phosphate), which is reduced by a NADPH linked reductase to yield glutamate gamma-semialdehyde (in equilibrium with delta 1-pyrroline-5-carboxylate). In the present work we found that the kinase, in the absence or presence of the reductase (and in the absence of NADPH), catalyzes stoichiometric formation of 5-oxo-L-proline and Pi from L-glutamate and ATP, but catalyzes hydroxamate formation at only about 10% of the rate of ATP-cleavage. A new substrate of the kinase was found; thus, cis-cycloglutamate (cis-1-amino-1,3 dicarboxycyclohexane), a glutamate analog which cannot cyclize to form an analog of 5-oxoproline, interacts effectively with the kinase. The trans form of cycloglutamate does not interact with the kinase; only the cis form can assume a diequatorial conformation equivalent to the extended conformation of glutamate. cis-Cycloglutamyl phosphate formation was shown and evidence was obtained for formation of an enzyme-ADP-cycloglutamyl phosphate complex. Although cis cycloglutamyl phosphate is not a reducible substrate of the NADPH-dependent reductase, the findings indicate that it interacts with the reductase. These studies, which elucidate several aspects of the mechanism of the utilization of glutamate for formation of delta 1-pyrroline-5-carboxylate, support the hypothesis that the kinase and reductase function as an enzyme complex. A model is suggested in which gamma-glutamyl phosphate formed on the kinase interacts with the reductase to form a gamma-glutamyl-reductase complex, which is reduced by NADPH to yield glutamate gamma-semialdehyde. PMID- 2567736 TI - Hypobetalipoproteinemia due to an apolipoprotein B gene exon 21 deletion derived by Alu-Alu recombination. AB - We report the molecular defect in an individual with homozygous hypobetalipoproteinemia. A unique TaqI restriction fragment length polymorphism was found in the midportion of the apolipoprotein B (apoB) gene using the genomic probe, pB51. The probe, which identifies TaqI fragments of 8.4 and 2.8 kilobases (kb) in normal individuals, hybridized to a single 11-kb fragment in the proband. The parents of the proband showed all three TaqI fragments, implying that they are heterozygotes for the mutant apoB allele. In this family, the mutant allele cosegregated with low total cholesterol levels and formal linkage analysis gave a decimal logarithm of the ratio score of 3.3 at a recombination frequency of 0. The polymorphic TaqI site was localized to an EcoRI fragment of 4 kb in normal individuals. The corresponding fragment in the proband was 3.4 kb, suggesting a 0.6-kb deletion in the mutant allele. Both the normal 4-kb EcoRI fragment and the mutant 3.4-kb EcoRI fragment were cloned and sequenced. In the normal allele, the 4-kb EcoRI fragment extends from intron 20 to 23. Exon 21 is flanked by Alu sequences that are in the same orientation. The mutant allele had a 694-bp deletion in this region which included a small part of the Alu sequence in intron 20, the entire exon 21, and most of the Alu sequence in intron 21. The polymorphic TaqI site, which lies within the Alu sequence in intron 21, was absent in the proband as a result of the deletion. The deletion of exon 21 results in a frame shift mutation and the introduction of a stop codon. Translation of the encoded mRNA would yield a prematurely terminated protein. This mutant apoB protein would be 1085 amino acids long with the 73 carboxyl terminal residues out of frame. We postulate that the deletion of exon 21 is the consequence of a crossover event between the Alu sequences in introns 20 and 21 resulting in nonreciprocal exchange between two chromosomes. PMID- 2567738 TI - Growth, enzyme activity, sugar transport, and hormone supplement responses in cells cloned from a pig kidney cell line LLC-PK1. AB - Three clones of the pig kidney cell line LLC-PK1 were isolated and characterized with regard to morphology, growth, proximal tubule enzyme activity, sugar uptake capacity, and hormone and drug responsiveness in a defined medium. Clone N4 was similar in morphology to the wild type (WT), whereas clone F8 showed loose attachment to the substrate, formed large, sweeping domes, and had an elongated desmosome junction between cells. The third clone, F2, did not form domes and showed a marked reduction in growth rate. Cultures of WT, N4, and F8 had higher specific activities of the enzyme alkaline phosphatase and gamma-glutamyl transpeptidase at confluence relative to growing cells; however, there was no evidence of an increase in activity of either enzyme at confluence in F2. Phlorizin-sensitive alpha-methyl-D-glucoside uptake and cytochalasin B-sensitive 2-deoxy-D-glucose uptake were measured in confluent cultures grown on porous filter supports. None of the clones lacked either of the hexose transport systems, although quantitative differences were evident. N4 cells grown in a defined medium in 96-well culture plates were tested in situ for their enzyme responses to differentiation inducers, tumor promoters, and hormones. Alkaline phosphatase activity was significantly increased at confluence by serum, parathyroid hormone (PTH), and vasopressin (AVP), and was decreased by tetradecanoylphorbol acetate (TPA) and epinephrine (EPI). Glutamyl transpeptidase activity was decreased at confluence by serum, TPA, and EPI. Similar tests on alpha-methyl-D-glucoside uptake showed that serum, TPA, PTH, and AVP had no significant effect on phlorizin-sensitive uptake; however, calcitonin increased uptake by 84% (n = 18). It was concluded that LLC-PK1 clones maintained in a defined medium are useful models for studying renal cell function. PMID- 2567737 TI - Concentration of transferrin receptor in human placental coated vesicles. AB - Coated vesicles were purified from human placenta by sedimentation, isopycnic centrifugation, and gel filtration. Quantitative Western blotting of the endogenous transferrin receptor (tfR) demonstrated the presence, on average, of roughly one receptor per vesicle. TfR appeared undersaturated with transferrin. After solubilizing vesicles in nonionic detergent, we looked for evidence of tfR interactions with other proteins. Solubilized tfR had an unexpectedly high mobility by gel filtration, apparently resulting from its self-association. This property was not seen in purified tfR or in tfR from a different cell fraction. The tfR complexes, though noncovalent, were largely resistant to conditions that disassemble coat proteins, and they did not appear to contain any other protein species. PMID- 2567739 TI - Pharmacokinetics of acrivastine after oral and colonic administration. AB - Six healthy male volunteers participated in this randomized, crossover open-label pharmacokinetic study consisting of two dosing segments separated by a washout period of at least 5 days. During each dosing segment, each volunteer received 12 mg of acrivastine, an investigational histamine H1-receptor antagonist, in a syrup form either orally or by colonic administration in random order. After oral and colonic administration, respectively, the following mean +/- SD pharmacokinetic parameters were obtained: Cmax 179 +/- 11 and 13.8 +/- 5.2 ng/ml; tmax, 0.85 +/- 0.13 and 3.60 +/- 0.56 hr; AUC0-12 hr, 576 +/- 57 and 104 +/- 46 hr.ng/ml. Differences between the oral and colonic administration for all three parameters were statistically significant (P less than 0.001). The mean +/- SD relative bioavailability of acrivastine from colonic compared to oral dosing was 0.18 +/- 0.09. It may be concluded, therefore, that appreciable absorption of acrivastine from the colon does not take place. These results suggest that comparison of pharmacokinetic profiles of some drugs after oral and colonic administration may be a useful technique for predicting bioavailability from a sustained release oral formulation. PMID- 2567740 TI - Cardiovascular effects of H2-receptor antagonists. AB - The type II histamine receptor antagonists, cimetidine and ranitidine, widely used in treatment of peptic ulcer disease have been reported to cause bradycardia. To evaluate the cardiovascular effects of H2 antagonists nineteen healthy volunteers were entered into a double-blind crossover comparison of cimetidine 300 mg qid, ranitidine 150 mg bid, and placebo. Subjects ingested study medicine for 7 days prior to being tested by the Bruce Exercise Test. Heart rate, blood pressure, oxygen consumption, expiratory volume, and fractional expiration of CO2 and O2 were measured at rest, exercise and recovery. A plasma sample for determination of cimetidine and ranitidine levels were obtained prior to the exercise period. Multivariate analysis and paired t test revealed no significant differences for the cardiovascular or pulmonary variables. However, in 5 subjects, the heart rate at 25% maximum VO2 was depressed 8% (P less than or equal to 0.03). This effect in a small percentage of the population suggests that further studies are needed to determine if subpopulations are affected. PMID- 2567741 TI - Rebound insomnia: a critical review. AB - Rebound insomnia, a worsening of sleep compared with pretreatment levels, has been reported upon discontinuation of short half-life benzodiazepine hypnotics. This paper reviews the existing sleep laboratory studies for the presence or absence of rebound insomnia following treatment with triazolam, temazepam, and flurazepam in insomniac patients or poor sleepers and, when possible, in normals. The results indicate that rebound insomnia is a distinct possibility after discontinuation of triazolam in both insomniacs and normal controls. Compared with baseline, disturbed sleep was reported in insomniacs or poor sleepers for the first 1 or 2 nights of withdrawal in seven of nine polygraphically recorded sleep studies following triazolam 0.5 mg and in one of two studies following triazolam 0.25 mg. In one study conducted in normal volunteers, rebound insomnia was observed following triazolam 0.5 mg but not triazolam 0.25 mg. In another study, which used subjective reports of sleep rather than polygraphic recordings, rebound insomnia was significantly attenuated after triazolam 0.5 mg by tapering the dose over 4 nights. The risk of rebound insomnia after temazepam 15 or 30 mg was low. In keeping with its long elimination half-life, flurazepam (30 mg) continued to exert beneficial effects for the first 2-3 withdrawal nights, but the possibility of a mild rebound insomnia cannot be dismissed during the intermediate withdrawal period (nights 4-10) following prolonged, consecutive, nightly administration (more than 30 nights). The benzodiazepine hypnotics are generally preferred over other types (barbiturates or nonbenzodiazepine, nonbarbiturate), but there are advantages and disadvantages related to half-life of the benzodiazepines. The risk of rebound insomnia is greater with the short half-life as compared with the long half-life benzodiazepines. PMID- 2567742 TI - A preliminary dose-ranging trial of proglumide for the treatment of refractory schizophrenics. AB - A dose-finding study of proglumide added to neuroleptics for the treatment of schizophrenic patients who were relatively refractory to their ongoing neuroleptic regimen was performed. Initially, four patients were open-label treated using a regimen of progressively increasing doses (proglumide 0.5-1024 mg/day) for 4 weeks. Afterwards, seven patients were given low doses (0.5 mg/day) followed by higher doses (500 mg/day) for a total period of 8 weeks. Overall, no improvement was seen in these refractory patients as a group at any dose. In individual patients, modest improvement or worsening of psychotic symptoms was observed. The results suggest that more potent cholecystokinin (CCK) antagonists and a greater knowledge of CCK pharmacology are needed before novel treatments exploiting the interaction of CCK and dopamine in the brain can be developed. PMID- 2567743 TI - Ultrastructural analysis of somatostatin-immunoreactive neurons and synapses in the temporal and occipital cortex of the macaque monkey. AB - Somatostatin-containing neurons and terminals have been analyzed in monkey temporal and occipital cortex by using light and electron microscopic immunohistochemistry. An antibody against Somatostatin-28, that was shown previously preferentially to label fibers (Morrison et al.: Brain Research 262:344-351, 1983), was utilized. As expected, few cell bodies were labeled. At the electron microscopic level, labeled cells presented a characteristic asymmetric position of the nucleus and very few symmetric or asymmetric synapses on the somatic surface. In all areas examined, somatostatin fibers formed a dense plexus in the most superficial layers (I-upper III). The density of labeled fibers in intermediate (deep III-IV) and deep layers (V-VI) varied considerably among areas. The synaptic relationships of the immunoreactive fibers were analyzed and postsynaptic targets quantified in V1, V2, and the superior and inferior temporal gyrus (STG and ITG, respectively). The synapses formed by somatostatin-labeled boutons were of the symmetric type (type II) and the primary postsynaptic targets were dendritic shafts. No regional differences were found in the distribution of the postsynaptic targets in layers I-upper III. The pattern of synapses in the deep layers was examined in STG. The frequency and distribution of postsynaptic targets was similar to the superficial layers of STG and the other temporal and occipital regions. In intermediate layers of the temporal cortex areas there was an increase in the proportion of synapses on dendritic spines. In a correlated light and electron microscopic analysis we examined synapses made by radial fibers in these regions and found that although the main targets are distal dendritic shafts, almost 40% of synapses were on dendritic spines. We suggest that the radial fibers may originate from a specialized interneuron, previously described as the double bouquet cell, and that this particular subset of somatostatin-containing double bouquet cells is likely to exhibit a very high degree of regional heterogeneity with a preference for association cortices with extensive corticocortical convergence. PMID- 2567744 TI - Distribution of sympathetic preganglionic neurons and monoaminergic nerve terminals in the spinal cord of the rat. AB - A study was made of the distribution of sympathetic preganglionic neurons identified by retrograde labeling with horseradish peroxidase from various peripheral nerve trunks and of the distributions of monoaminergic terminals in the spinal cord of the rat. Nerve terminals were stained immunohistochemically by using antisera raised against tyrosine hydroxylase, phenylethanolamine-N-methyl transferase, neuropeptide Y, and 5-hydroxytryptamine and by using formaldehyde induced fluorescence. The three-dimensional distribution of sympathetic preganglionic neurons was described by using computer reconstruction and compared with the arrangement of each population of immunohistochemically stained terminals in the intermediate zone. Although monoaminergic terminals are associated with most sympathetic neurons, particularly in the intermediolateral column, the relationship of many terminals to sympathetic neuron somata in other parts of the intermediate zone is tenuous. Some of the descending innervation may terminate on interneurons. The data are consistent with the coexistence of phenylethanolamine-N-methyl-transferase and neuropeptide Y in terminals arising from cell bodies in the C1 region in the ventrolateral medulla and with the presence of at least two populations of catecholaminergic terminals as well as the adrenergic one. Serotoninergic terminals are denser and have a different arrangement from those of catecholaminergic terminals in the intermediate zone. PMID- 2567746 TI - The International Council of Nurses. PMID- 2567745 TI - Generalization of therapeutic changes in agoraphobia: the role of perceived self efficacy. AB - Investigated the extent and mechanisms of therapeutic generalization across distinct areas of agoraphobic dysfunction. Twenty-seven severe agoraphobics were each given performance-based treatment for some phobic areas while leaving their other phobias untreated. Behavioral tests revealed that (a) the treated phobias improved significantly more than the untreated (transfer) phobias, (b) the transfer phobias improved significantly more than control phobias, and (c) the transfer benefits were highly variable within and between subjects. Analyses of possible cognitive mechanisms revealed that perceived self-efficacy accurately predicted treatment and transfer effects even when alternative factors such as previous behavior, anticipated anxiety, anticipated panic, perceived danger, and subjective anxiety were held constant. In contrast, these alternative factors lost most or all predictive value when self-efficacy was held constant. Agoraphobia thus appears to be neither a unitary entity nor a mere collection of independent phobias, but a complexly patterned problem governed largely by self perceptions of coping efficacy. PMID- 2567747 TI - Drug treatment of functional dyspepsia. A meta-analysis of randomized controlled clinical trials. AB - The results of therapeutic trials in functional dyspepsia (FD), a frequently encountered condition, are contradictory. Our aim, then, was to produce a pooled estimate, or meta-analysis, of a series of short-term randomized placebo controlled clinical trials on the pharmacological treatment of FD with antisecretory and gastrokinetic drugs. We retrieved trials for analysis purposes by consulting computerized data bases and by scanning published reviews, Current Contents, and references cited in the individual studies. We also requested bibliographical updates from the medical departments of the manufacturers of the drugs used in the various trials. Of 74 trials retrieved by these means, 23 proved eligible for meta-analysis on the basis of six selection criteria defined a priori. Results were expressed in terms of "therapeutic success" (TS), which includes "symptom-free patients," patients with "significant improvement in symptoms," "excellent results," and so on. The differences in TS rates between the various drugs and placebo were calculated in each trial as the algebraic difference together with the respective 95% confidence interval (95% C.I.); the pooling of results of all eligible trials was done using Cochran's weighted method. With antisecretory drugs, the mean difference in TS rates versus placebo was +20% (95% C.I.: 14-24%). The therapeutic gain for the respective antisecretory agents was 25% (95% C.I.: 14-36%) in the case of pirenzepine and 18% (95% C.I.: 12-24%) in the case of H2 antagonists. Meta-analysis of trials with gastrokinetic drugs also showed superior efficacy of these agents compared with placebo, with a mean difference in TS rates of +46% (95% C.I.: 40 52%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2567748 TI - Domains of medical practice: physicians' assessment of the role of physician extenders. AB - The relationship between physicians and physician extender occupations is considered in light of the stratification of medical care workers. Professional dominance between various medical occupations and professional competition among physicians are considered. Data from a national sample of physicians taken in 1981 are used to determine 1) under what conditions physicians are likely to consider the employment of physician extenders helpful for improving quality of care and 2) which characteristics and available resources are associated with the belief that the employment of physician extenders will improve medical care. Of four different service populations considered, physicians are most likely to think that the use of physician extenders to care for the urban poor will aid care and are least likely to think that their employment will improve care for obstetrics and pediatrics cases. The results indicate that physicians' beliefs regarding the employment of physician extenders are dependent on the clientele as well as on the relative position of the two occupational groups in the hierarchy of medical care occupations. PMID- 2567749 TI - Puncture wounds to the foot. AB - Puncture wounds to the foot are common presenting problems in most busy emergency departments. Although seemingly benign, the sequelae after simple puncture wounds to the foot can include cellulitis, retained foreign bodies, or even osteomyelitis. Inadequate scientific research on this topic has left only anecdotal or retrospective reports for review in the medical literature. Antibiotics, radiographs, or surgical exploration in the management of puncture wounds to the foot all lack clinical studies to support their use. This review summarizes the literature and points to the inconsistencies in the management of puncture wounds to the foot. PMID- 2567750 TI - Standardization of disinfectant testing. PMID- 2567751 TI - Surveillance and use of computers in hospital infection control. PMID- 2567752 TI - A comparison of hypochlorite and phenolic disinfectants for disinfection of clean and soiled surfaces and blood spillages. AB - Suspension test methods were used to compare phenolic and hypochlorite disinfectants under conditions as recommended for use in hospitals. Using plasma to simulate soiled conditions, Clearsol (1% v/v) and Stericol (2% v/v) produced satisfactory disinfection, (i.e. 5 log reduction in 5 mins against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa in the presence of up to 50% v/v plasma (25 mg ml-1 plasma protein). Although sodium dichloroisocyanurate (NaDCC) at 2500 ppm AvCl2 was effective in the presence of up to 20% plasma, compared to 10% plasma for sodium hypochlorite 2500 ppm, sensitivity of NaDCC to inactivation was greater than for phenolics. In the presence of blood, both hypochlorites and phenolics were substantially inactivated, although here the effectiveness of NaDCC at 10,000 ppm was equivalent to the phenolics. Our results indicate that hypochlorites at 10,000 ppm, Clearsol 1% and Stericol 2% may be ineffective for treatment of blood spills unless applied at v:v ratios of 9 parts disinfectant to 1 part blood. In this situation, chlorine-releasing powder formulations, which produce higher AvCl2 concentrations and contain spilled material, offer an effective alternative. PMID- 2567753 TI - Use of sodium dichloroisocyanurate granules for spills of body fluids. AB - The use of chlorine-containing granules for disinfecting body fluid spills has been evaluated by hospital trials and laboratory tests. Hospital trials were carried out by nurses using 'Presept' disinfectant granules according to a protocol. In general they preferred using granules to bleach and, in 50 tests using granules on natural and artificial spills in wards, no organisms were recovered from the floor by contact plates after using the granules. Laboratory tests were carried out on 'Haz-Tab' granules, 'Biospot' disinfectant powder, 'Presept' disinfectant granules, 'Virusorb' absorbent powder and 'Titan' Sanitizer SU 357 using a standardized surface test. Available chlorine levels varied from 57.8% to 1.0% and the performance of products in the surface test varied with the chlorine level present. Granules containing a relatively high level of chlorine have the advantages that spilled material is contained and that a contact time of only 2-3 min is required before the spill can be safely removed. PMID- 2567754 TI - Contamination of subclavian vein catheters: an intraluminal culture method. AB - An intraluminal culture method was evaluated for central venous catheters and compared to conventional catheter tip cultures and cultures from the insertion site. The colonization-rate of the catheters was 43.7% using conventional tip culture, 34.4% with the intraluminal method and 40.1% at the skin puncture site. Only 37.5% of the catheters showed identical bacteria at the skin puncture site and catheter-tip, compared with 87.5% between intraluminal culture and catheter tip culture (P less than 0.05). PMID- 2567755 TI - Randomised study of prophylactic parenteral sulbactam/ampicillin and cephazolin in biliary surgery: significant benefit in jaundiced patients. AB - Two hundred consecutive patients undergoing biliary-tract surgery were entered into a randomized trial of prophylactic single dose cephazolin or sulbactam/ampicillin. There was no overall difference in the infection rates between the two antibiotic groups, but in the group of patients with jaundice there was an excess of wound infections in the cephazolin group compared to the sulbactam/ampicillin group (35% vs. 14%). We conclude that sulbactam/ampicillin is a satisfactory prophylactic agent for use in biliary-tract surgical sepsis, and that it may be superior to cephazolin in jaundiced patients. PMID- 2567756 TI - The effect of incisional plastic drapes and redisinfection of operation site on wound infection following caesarean section. AB - In a randomized prospective multicentre study of post-caesarean wound infection among 1,340 women in eight hospitals, the effect of redisinfection of the skin around the incision before skin closure and the effect of adhesive skin drapes were investigated. The overall rate of wound infection with pus was 5.0% (range 3.5-8.9%). The study showed a reduction in postoperative wound infection associated with redisinfection (P = 5.5%), while no benefit from adhesive plastic drapes could be demonstrated. PMID- 2567757 TI - The DANOP-DATA system: a low-cost personal computer based program for monitoring of wound infections in surgical ward. AB - A low-cost personal computer program to monitor surgical wound infections was developed in parallel to the Danish national guidelines for recording postoperative wound infections. Internationally accepted definitions were used. The program offers three fixed-data entry screens and produces user-specified variations of four standard tables, comprising most of the epidemiological data needed for surveillance and infection control. The program was tested in Danish hospitals and was found to serve well as a simple local tool for the operating staff, offering fast information on infection rates. Results from two hospitals consisting of 3904 operations are presented. Infections occurring after discharge were included. Overall infection rates for clean wounds were 2.3%, clean contaminated wounds 4.7%, contaminated wounds 4.3% and dirty operations 8.3%. None of the hospitals had used infection surveillance systems before. PMID- 2567758 TI - Endemicity of Legionella pneumophila serogroup 3 in a hospital water supply. AB - A microbiological and epidemiological investigation at the Infectious Diseases Hospital in Turin, Italy, demonstrated Legionella pneumophila serogroup 3 at 10(2) to greater than 4 X 10(3) cfu l-1 from 24 of 32 hot water samples collected from hand-basins in six separate buildings. A sample taken from the public water supply, and a hot water sample (80 degrees C) collected from hot water tanks, did not yield legionellas. Legionella pneumophila serogroup 3 was found in samples taken at the first point of mixed hot and cold water (50 degrees C) at 3 X 10(2) cfu l-1. 12 of 26 samples from the shower-heads yielded 10(3) to 2.5 X 10(5) cfu l-1 and one of 12 water samples from oxygen bubble humidifiers tested yielded 1.6 X 10(4) cfu l-1. No other legionellas species or serogroups of Legionella pneumophila were isolated during the study. No cases of nosocomial pneumonia were detected among 3653 patients' records, nor was there serological evidence of Legionella infection in the 180 patients tested. PMID- 2567759 TI - Nosocomial Legionella micdadei pneumonia: 10 years experience and a case-control study. AB - Sixteen patients with nosocomial Legionella micdadei pneumonia, diagnosed between 1977 and 1988, were studied retrospectively to define clinical and epidemiological characteristics of the disease. Also, a case-control study was performed comparing the five patients with L. micdadei pneumonia during a cluster of cases in 1982, with uninfected patients with the same underlying diagnoses. No significant differences were noted in the case-control study with regard to age, presence of leucopenia, intensity or duration of immunosuppressive therapy, bed location, duration of hospital stay, frequency of transplant rejection or overall mortality. Legionella micdadei isolates from a sink on the renal transport ward, from hot water storage tanks, and one clinical isolate had identical cellular fatty acid composition. Extensive sampling of other potential sources failed to yield the organism. This indirect evidence suggests potable water as the source of infection. PMID- 2567760 TI - Nosocomial Branhamella catarrhalis in a paediatric intensive care unit: risk factors for disease. AB - There have been few reports on Branhamella catarrhalis as a nosocomial pathogen, and no risk factors for nosocomial infection have been identified. We report 11 cases (mean age 22 months) of nosocomial Branhamella catarrhalis respiratory tract infection in a paediatric intensive care unit (PICU) over a two-year period. There were 2 cases of pneumonia and 9 cases of bronchitis. Branhamella catarrhalis was the sole isolate recovered in 6 cases and was associated with other respiratory pathogens in 5 cases. A case-control study with two age-matched controls per patient (mean age 24.1 months) was undertaken to identify potential risk factors for infection; risk factors identified were the presence of an endotracheal tube (p less than 0.02) and frequent endotracheal tube suction (p less than 0.05). Five of 6 tested strains from PICU patients produced beta lactamase. DNA preparations of 4 B. catarrhalis isolates from PICU patients revealed no plasmids. B. catarrhalis should be considered a potential nosocomial pathogen. PMID- 2567761 TI - Outbreak of Clostridium difficile diarrhoea in an orthopaedic unit: evidence by phage-typing for cross-infection. AB - In a three-week period five patients had diarrhoea in an orthopaedic unit. The first case was clinically diagnosed as pseudomembranous colitis but the causative agent was not sought. Of the remaining cases, two were Clostridium difficile positive. The outbreak then apparently ceased, but during the following several days two of seven stool samples taken at random from asymptomatic patients yielded C. difficile. Phage-typing of the isolates showed that all apparently belonged to the same strain. PMID- 2567762 TI - Legionella pneumophila serogroup 12 pneumonia in a renal transplant recipient: case report and environmental observations. AB - We describe the first reported case of pneumonia due to Legionella pneumophila serogroup 12 in the UK. This hospital-associated infection occurred in an immuno incompetent patient and coincided with a change in character of the local environmental strains of legionellas. The patient produced a serological response both to her own isolate and to L. pneumophila serogroups 1-6. Thus serodiagnosis was attainable using the usual screening antigens. PMID- 2567763 TI - Cost-effectiveness in hospital infection control--lessons for the 1990s. PMID- 2567764 TI - An outbreak of puerperal fever caused by group C streptococci. AB - Between 19 February and 18 April 1987, 33 confirmed cases of puerperal fever caused by Streptococcus equisimilis serotype T204 occurred at three hospitals in and around Chelmsford. Most of the cases (70%) occurred on one ward, in which toilet seats and a shower are believed to have aided transmission, although insufficient data were obtained to exclude a role for person-to-person spread. Possession of M-protein antigen was demonstrated in the outbreak strain. PMID- 2567765 TI - Infection following caesarean section. AB - In a prospective clinical study of post-operative infection in 124 patients undergoing Caesarean section, 39 (31.5%) patients developed a total of 45 infections. There was no significant difference in infection rates between elective and emergency procedures. Five patients (4%) developed endometritis, wound infection was found in 14 (11.3%) and 18 patients (14.5%) developed a post operative urinary tract infection. Pre-operative C-reactive protein levels and intraoperative swabs of the uterine cavity were not helpful in the early detection of endometritis. The risk factors predisposing to post-caesarean infection were obesity and low socioeconomic status. PMID- 2567766 TI - Methicillin-resistant Staphylococcus aureus in two burn units: clinical significance and epidemiological control. AB - Routine isolation adequately controlled MRSA strains in two burns units with a restrictive antibiotic policy. Ventilation control and more rigorous change of clothing offered no further advantage. No carriers among staff were found, but some suffered minor skin lesions that were the source of further MRSA spread. Spread of MRSA from the unit to other parts of the hospital was prevented by early identification of colonized patients and by restricting patient and staff movement. PMID- 2567767 TI - RNA-electrophoresis as a typing method for nosocomial rotavirus infection in a special-care baby unit. AB - During January and February 1988 an outbreak of nosocomial rotavirus infection occurred in a special-care baby unit (SCBU). Seven infants were affected, of whom five had symptoms of diarrhoea. Typing by RNA electrophoresis (electropherotyping) demonstrated that a single rotavirus strain was responsible for the outbreak. The epidemic electropherotype was introduced by an infant with diarrhoea admitted directly into the SCBU from the community. Eight other electropherotypes were identified during January and February in patients with community-acquired rotavirus diarrhoea on other paediatric wards, but the outbreak strain occurred only on the SCBU. Electropherotyping is a useful and rapid method for tracing the epidemiology of hospital-acquired rotavirus infection. PMID- 2567768 TI - Cleansing of hands with emulsion--a solution to skin problems of hospital staff? AB - In this study of hand cleansing, nursing staff with skin problems used emulsion followed by rinsing with water, while control groups, with or without skin problems, used liquid soap and water. Clinical evaluation of the without skin problems, used liquid soap and water. Clinical evaluation of the skin by a dermatologist, as well as by self-assessment, suggested that when the staff with skin problems used emulsion their skin was in better condition than that of controls with similar skin problems. This favourable effect on skin was later confirmed in extended use of emulsion for hand cleansing in other hospital wards. Objective evaluation of skin condition with biophysical measurements was unsuccessful. Emulsion cleansing of the hands seems to offer a favourable alternative to washing with soap and water, especially for persons with skin problems. PMID- 2567769 TI - Postoperative wound infection: relation to different types of operation and wound contamination categories in orthopaedic surgery. AB - During one year, 1,407 operations were performed on inpatients in an orthopaedic department. The presence or absence of postoperative wound infection was recorded during the hospital stay, at discharge and at follow up. Wound infections were linked to operations of different contamination categories and to the different types of operation performed. Better information about the risk for postoperative wound infection and the need for antibiotic prophylaxis was provided by considering the different operation types. PMID- 2567770 TI - Prophylactic parenteral cefuroxime: subcutaneous concentrations in laparotomy wounds. AB - Plasma and subcutaneous adipose tissue cefuroxime concentrations were measured in laparotomy wounds, by means of high-pressure liquid chromatography, in 12 patients undergoing elective abdominal operations. After intravenous administration of 1.5 g cefuroxime at induction of anaesthesia, the measured concentrations in serum and wound tissue during a 2 h period were above the MIC 90 of most micro-organisms derived from the alimentary tract. Tissue peak levels were reached within 15 min and the tissue half life was 1.5 h. PMID- 2567771 TI - Evaluation of the Draeger Anaesthetic Equipment Washing Machine (ANDA 9002). AB - An evaluation was made of an anaesthetic equipment washing machine. Our results largely substantiated the manufacturers claims. A clean, dry, disinfected load could be produced within 75 min. After minor modifications were made to the machine by the manufacturers, potential users were satisfied with its performance. We recommend consideration of this type of equipment for use in hospitals. PMID- 2567772 TI - Delayed thymocyte maturation in the trisomy 16 mouse fetus. AB - Mouse fetuses with trisomy 16, an animal model for human trisomy 21 (Down syndrome), have severe defects in several hematopoietic stem cell populations and a marked reduction in thymocyte number. To determine whether there are other defects in the development of the trisomic thymus, the ontogeny of the cell surface antigenic determinants, Thy-1, Ly-1, CD3, CD4, CD8, and TCR v beta, was investigated. The trisomy 16 thymocytes were able to express all of determinants either during fetal life (days 14 to 19 of gestation) or in cultures of intact thymus lobes. However, in all instances (except for Thy-1, which already had a high proportion of expressing thymocytes by day 14), there was a delay in the time at which the determinants were first expressed, as manifested by reduced numbers of positively staining cells. Furthermore, there was also a delay in the rate at which the positively staining cells attained maximal Ag densities. Overall, there was an approximate 2 day lag in development of the fetal trisomic thymocytes. This lag permitted the identification of a large population of CD4-8+ cells prior to the appearance of CD4+8+ thymocytes. These findings are consistent with the identification of CD4-8+ as an intermediate stage between CD4-8- and CD4+8+ in fetal thymocyte ontogeny. PMID- 2567773 TI - Characterization of allelic V kappa-1 region genes in inbred strains of mice. AB - Germ line genes encoding mouse Ig kappa-chains belonging to the V kappa-1 group have been isolated from BALB/c, NZB, and CE, three inbred strains of differing kappa haplotype. The V kappa-1A and V kappa-1C germ line genes isolated from BALB/c (Ig kappa c) were identical to those previously described. These are the two major V kappa-1 germ line genes in BALB/c and together account for 40 of the 53 expressed V kappa-1 sequences that have been reported to date. Allelic differences in a single germ line variable region gene (V kappa-1A) in different strains of mice explain the differences in L chain IEF patterns previously associated with the Ig kappa-Ef2 locus. The rearranged kappa-gene expressed in the BALB/c myeloma MOPC-460 has been isolated and found to represent a V kappa-1A somatic variant differing by three nucleotides from the germ line V kappa-1A gene. Germ line genes isolated from NZB (Ig kappa b) and CE (Ig kappa f) show greater than 95% identity with the BALB/c genes over the 1700 nucleotides compared. Comparison by region indicated the greatest conservation of sequence occurs in and around the leader exon followed by the V-region exon. The NZB gene encodes the amino acid sequence found in the myeloma PC-2205, previously designated V kappa-1B. The V kappa-1 gene isolated from CE is likely an allele of the BALB/c V kappa-1C gene as the two share greater than 96% identity over 1700 nucleotides. The CE gene has been designated V kappa-1Cf. Ancient remnants of LINE-1 repetitive elements were detected approximately 400 bp downstream of all of the V kappa-1 genes. These possess greater homology with repetitive elements found near other kappa genes than they do with the native L1Md sequence. PMID- 2567774 TI - Polymorphisms in human H chain V region genes from the VHIII gene family. AB - Polymorphisms of the Ig H chain V region (VH) genes were examined with probes from the coding and flanking regions of a gene from the largest VH gene family, VHIII. The 5'-flanking probe gave the simplest pattern and revealed the largest number of polymorphic fragments. Analysis of unrelated individuals and of families identified five polymorphic loci. Two alleles were detected for each of two of the loci, whereas a polymorphic band was scored as present or absent for the other three loci. The polymorphic fragments segregated in the expected Mendelian fashion and parental haplotypes could be assigned in all cases. Comparison of the patterns obtained with the flanking and coding region probes suggests that the human VHIII gene family is highly polymorphic and may contain several hundred V genes. This method, as well as the polymorphism detected, can be used to investigate the organization and germ-line variation of H chain V genes and their inheritance in normal individuals and in individuals with immunologic disorders. PMID- 2567775 TI - Crypticola clavulifera gen. et sp. nov. and Lagenidium giganteum: oomycetes pathogenic for dipterans infesting leaf axils in an Australian rain forest. AB - The isolation of two entomopathogenic fungi from Forcipomyia marksae larvae collected in leaf axils of Colocasia macrorrhiza in northeastern Queensland rain forests is reported. An oomycete, in which the zoospores complete their development wholly within the sporangium and are discharged through a short papillar extension, is described as a new genus and species, Crypticola clavulifera. A strain of the well-known oomycete, Lagenidium giganteum, was isolated from F. marksae--the first record of this fungus infecting a member of the Ceratopogonidae. The pathogenesis and zoosporogenesis of the two fungi in mosquito and ceratopogonid larvae are described. PMID- 2567776 TI - [A new diagnostic method of lung cancer by DNA-RFLP]. PMID- 2567777 TI - Crohn's disease, sulfasalazine, and mycobacteria. AB - The in vitro antimycobacterial activities of sulfasalazine and its split products, mesalazine (5-aminosalicylic acid) and sulfapyridine, against 170 mycobacterial strains belonging to 17 species were compared with those of diaminodiphenylsulfone, sulfadimethoxine, olsalazine, and para-aminosalicyclic acid. Only para-aminosalicylic acid showed consistent activity against Mycobacterium tuberculosis. These data suggest that the principle of activity of sulfasalazine in Crohn's disease is not an antimycobacterial effect. PMID- 2567778 TI - Intracerebral drug delivery in rats with lesion-induced memory deficits. AB - Pharmacological treatments directed at increasing cortical acetylcholine activity in patients with Alzheimer's disease have largely been disappointing, perhaps because denervated areas of brain may not be exposed to adequate amounts of drug. A new method has been developed to enable localized intracerebral delivery of neurotransmitter substances using a polymeric drug delivery system. Microspheres of a polyanhydride sebacic acid copolymer were impregnated with bethanechol, an acetylcholinesterase-resistant cholinomimetic. Twenty rats received bilateral fimbria-fornix lesions, producing cholinergic denervation of the hippocampus and marked impairment in spatial memory. The animals were trained for 2 weeks to run after which they received bilateral intrahippocampal implants of saline (five rats), blank polymer (five rats), or bethanechol-impregnated polymer (10 rats). Following implantation, spatial memory was assessed by radial-maze performance testing for 40 days. Untreated lesioned rats showed persistently poor spatial memory, entering maze arms with near random frequency. Similarly, animals treated with saline and blank polymer did not improve after implantation. Rats treated with bethanechol-impregnated microspheres, however displayed significant improvement within 10 days after implantation; this improvement persisted for the duration of the experiment (p less than 0.05, Student's t-test). Histological analysis of regional acetylcholinesterase staining showed widespread loss of activity throughout the hippocampus bilaterally in all animals. The microsphere implants were visible within the hippocampus, with minimal reactive changes in surrounding brain. It is concluded that intracerebral polymeric drug delivery successfully reversed lesion-induced memory deficits, and has potential as a neurosurgical treatment method for Alzheimer's disease and other neurodegenerative disorders. PMID- 2567780 TI - Epidural opioids in children. AB - Experience with spinal opioids in children is limited but is expanding. Anatomy, pharmacology, technique, and results are reviewed. Complications and side effects are described. PMID- 2567781 TI - Issues in the use of depot antipsychotics. PMID- 2567779 TI - D2-dopamine receptor specific brain uptake of carbon-11-labeled YM-09151-2. AB - The in vivo D2-receptor specific brain uptake of N-[(2RS,3RS)-1-benzyl-2- methyl 3-pyrrolidinyl]-5-chloro-2-methoxy-4-[11C]methylaminobenzamide ([11C]YM-09151-2), was investigated. In rat brain the high uptake of [11C]YM-09151-2 in striatum was displaced with sulpiride, spiroperidol, and YM-09151-2. SCH-23390 and ritanserin, D1-dopamine and S2-serotonin antagonists, showed no effect on the distribution of [11C]YM-09151-2. In the striatum at 60 min, 95% of the radioactivity was detected as [11C]YM-09151-2 by high performance liquid chromatography. On the other hand, 41% of 11C in the plasma at 60 min was observed as metabolites. In vivo autoradiography showed a high uptake of [11C]YM-09151-2 in the striatum and in the nucleus accumbens of rat brain. A high uptake of radioactivity was also found in the canine basal ganglia with positron emission tomography. The uptake was reduced by pretreatment with spiroperidol. The present results demonstrate that [11C]YM-09151-2 is a D2 receptor specific compound and is a potential in vivo tracer for measuring D2 receptors. PMID- 2567782 TI - Highly selective kappa-opioid analgesics. 2. Synthesis and structure-activity relationships of novel N-[(2-aminocyclohexyl)aryl]acetamide derivatives. AB - This paper describes the chemical synthesis and the development of structure activity relationships (SAR) for the kappa opioid receptor affinity and mu/kappa opioid receptor selectivity of novel N-[(2-aminocyclohexyl)aryl]acetamide derivatives. The SAR of this series are investigated by consideration of structural modifications made to the aromatic moiety, the amide linkage, and cyclohexane and the pyrrolidine ring substituents of the prototype kappa selective agonist, PD117302 (trans-N-methyl-N-[2-(1 pyrrolidinyl)cyclohexyl]benzo[b]thiophene-4- acetamide) (1). The kappa and mu opioid receptor binding affinities of 23 novel compounds are reported. It is observed that optimal mu/kappa receptor selectivity is obtained with a benzo[b]thiophene aromatic system attached via the C-4 position, which is discussed in terms of steric and electronic parameters. The amide linkage has been replaced with the reversed amide, an ester, an aminomethylene, a thioamide, and a secondary amide. The best of these isosteres is the N-methyl amide. Substitution of the pyrrolidine ring of PD117302 in the 3-position with a hydroxymethylene group increases the mu/kappa selectivity compared to the unsubstituted compound, e.g. compound 14, trans-(+/-)-N-methyl-N-[2-[3 (hydroxymethyl)-1-pyrrolidinyl] cyclohexyl]-4-benzo[b]furanacetamide monohydrochloride, mu/kappa receptor selectivity = 244. The cis fused, 4,5 dimethyl ether substituted cyclohexane analogue trans-(+/-)-N-methyl-N-[4,5 dimethoxy-2-(1-pyrrolidinyl) cyclohexyl]-benzo[b]thiophene-4-acetamide monohydrochloride (32) has high in vitro kappa opioid receptor affinity (Ki = 16 nM) and equipotent analgesic activity to morphine after iv administration in rats. PMID- 2567783 TI - Heteroaromatic analogues of the alpha 2-adrenoreceptor partial agonist clonidine. AB - A 1,4-dioxane analogue (1) of the alpha 2-adrenoreceptor partial agonist clonidine (2) has previously been shown to possess an interesting but complex pharmacological profile. In this study, from a series of other heterocyclic analogues of clonidine, the 1,4-oxazines 6 and 12 were found to resemble 1 in that they are partial alpha 2-agonists in the periphery and are excluded from the central nervous system. However, when given directly into the brain, they behave as pure alpha 2-antagonists. PMID- 2567785 TI - Population frequencies of three DNA alleles linked to the Duchenne muscular dystrophy gene. AB - To enquire whether the known X linked probes linked to the Duchenne muscular dystrophy gene vary in their RFLP frequencies, three probes, 754, XJ1.1, and pERT87.8, were tested in European, Indian Muslim, and West African samples. Though the average heterozygosity for the three together is fairly similar in the three populations, significant differences in allele frequencies were evident. PMID- 2567784 TI - Clinical variability of osteogenesis imperfecta linked to COL1A2 and associated with a structural defect in the type I collagen molecule. AB - We report a family in which dominant osteogenesis imperfecta segregates with a COL1A2 haplotype and is associated with a structural defect in the helical region of the type I procollagen molecule. All affected subjects had short stature, dentinogenesis imperfecta, and myopia; however, great differences were observed in the number of fractures and in the degree of bone deformity. Identical biochemical changes were found in the type I collagen molecules synthesised by fibroblasts of subjects with severe or minimal bone fragility. These results confirm that mutations in the triple helical region of alpha 2(I) chains produce a milder phenotype than analogous mutations in the alpha 1(I) chains, but indicate that, in addition to defects in the type I collagen molecule, other factors may modulate the degree of bone involvement in osteogenesis imperfecta. PMID- 2567786 TI - Multiple endocrine neoplasia type II with Zollinger-Ellison syndrome caused by a solitary pancreatic gastrinoma. PMID- 2567787 TI - [P-fimbriated Escherichia coli and serum antibody responses to P-fimbriae in pregnant women with urinary tract infections and their children]. AB - Escherichia coli (E. coli) is still a major pathogen in urinary tract infections (UTI). It was found that 15 out of 20 cases (75%) of E. coli related UTI were caused by P-fimbriated E. coli, compared to the mere 15% of E. coli isolated from urine and 22% from the stool of healthy controls that were P-fimbriated. All patients studied were pregnant women and their delivered children. Antibody responses to P-fimbriae in the sera of these patients were detected with enzyme linked immunosorbent assay (ELISA) using purified P-fimbriae. Positive antibody responses were observed at titers of 800-6400 in 8 out of 9 cases of UTI of pregnant women caused by P-fimbriated E. coli. The high level of antibody titers persisted for one month on average and then decreased. These antibodies to P fimbriae were essentially IgG and transmitted to delivered children from UTI mothers. Therefore, the protective role of these antibodies from P-fimbriated E. coli infections in new born children has been suggested. PMID- 2567788 TI - [Cryptorchism and its surgical treatment]. AB - 545 operations were conducted in the clinic in various forms of cryptorchidism. Downward transposition of the testis after Keetley-Torek-Gertsen was the operation of choice. The author was the first to perform one-stage transposition of both testes with the use of microsurgical techniques in bilateral cryptorchidism. The modern principles of nonoperative and operative treatment of cryptorchidism are discussed and the necessity for early operative treatment (no later than 2 years) in substantiated. To study the results of treatment according to the form of cryptorchidism, localization of the testes, and the patients' age, the author examined the hypophyseal-testicular system and spermatogenesis in 80 patients with various forms of cryptorchidism. The study demonstrated the possibility of medical and social rehabilitation of patients in proper surgical tactics and, above al, in timely treatment. PMID- 2567789 TI - Preparation of the internal mammary artery graft. Which is the best method? AB - Early reports questioned the adequacy of flow of the internal mammary artery when used routinely as a bypass graft. "Adequate" mammary artery flow is now contested only in certain situations, that is, left ventricular hypertrophy, acute myocardial infarction, and reoperations. To compare the methods of mammary pedicle graft preparations with free mammary artery flow, we studied 31 patients who had the left internal mammary artery harvested for elective coronary artery bypass grafting. Group I comprised 14 patients whose mean body surface area was 1.91 m2. Systolic, diastolic, and mean arterial blood pressures, left atrial pressure, and heart rate were recorded and stabilized during flow measurements. Free flow of the internal mammary artery was measured before any pharmacologic manipulation and ranged from 5 to 44 ml/min (mean 18 ml/min). The grafts were sprayed and wrapped in sponges soaked in diluted papaverine solution (60 mg in 40 ml normal saline) for an average of 21 minutes. Free flow ranged from 10 to 108 ml/min (mean 51 ml/min). Intraluminal papaverine of the same dilution was then injected with hydrostatic dilatation. Immediate internal mammary artery flows rose from 150 to 333 ml/min (mean 229 ml/min). Group II comprised 17 patients who had internal mammary artery takedown under the exact conditions used in group I. Mean body surface area was 1.89 m2. Mammary artery pedicles were injected with diluted papaverine throughout their lengths with size 25 needles. After an average of 19.5 minutes, free flow ranged from 28 to 132 ml/min (mean 69 ml/min). Intraluminal diluted papaverine was then administered as in group I, and flows increased from 144 to 280 ml/min (mean 198 ml/min). The distal internal mammary arteries in both groups were 1.75 to 2.5 mm in internal diameter at the site of arteriotomy for flow measurement. This study shows that all mammary arteries are in spasm immediately after harvest and that flow is inadequate before any pharmacologic intervention. Although extraluminal vasodilators will increase free mammary artery flow, intraluminal papaverine followed by hydrostatic dilatation raises free flow to maximal capacity. Subsequent graft spasm has not been observed.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2567791 TI - Effect of long-term caloric restriction on brain monoamines in aging male and female Fischer 344 rats. AB - The present study examines the changes in central monoamines and their metabolites in aged male and female rats after long-term caloric restriction. Fischer 344 rats of both sexes (n = 5-10/group) were maintained on one of two dietary regimens: ad libitum NIH 31 diet or 60% by weight of the ad lib. intake (restricted), supplemented with vitamins and minerals. Animals received these diets from the age of 14 weeks until killed at 22.25 months of age. Caudate nucleus (CN), hypothalamus (HYPO), olfactory bulb (OB) and nucleus accumbens (NA) were assayed for content of norepinephrine (NE), dopamine (DA) and its metabolites (dihydroxyphenylacetic acid, DOPAC, and homovanillic acid, HVA) and serotonin (5-HT) and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) using HPLC/EC. Relative to the ad lib. group, restricted rats of both sex showed significant decreases in NE content in CN, HYPO and OB. DA and 5-HT content were decreased significantly in the CN and HYPO. No significant changes were found in the levels of DA metabolites in all brain regions studied. While the 5-HIAA level was significantly reduced in the HYPO and NA of the female restricted rats, it was increased several-fold in the OB of the male restricted animals. These preliminary results suggest that long-term caloric restriction alters brain monoamine concentrations, an effect which may in turn modify the normal rate of aging. PMID- 2567790 TI - Antagonism of leukotriene receptors and administration of a 5-lipoxygenase inhibitor do not affect hypoxic vasoconstriction. AB - The role of leukotrienes in hypoxic vasoconstriction remains controversial. Our previous study using the lipoxygenase inhibitor BW 755C in dogs failed to show a substantive role for leukotrienes in hypoxic vasoconstriction. To clarify further the role of leukotrienes, we designed 3 protocols. In the first protocol, we examined the effects of LTD4 boluses on the pulmonary circulation in 6 anesthetized dogs. LTD4, 1 microgram/kg, (a large dose relative to other species) produced no detectable constriction of the pulmonary artery, while systemic vascular resistance increased 41 +/- 17% (SD), left atrial pressure rose 3.5 +/- 1.5 mmHg, and cardiac output fell 18 +/- 8%. Two leukotriene receptor antagonists, LY171883 and L-648051, decreased these effects by more than 50%. In the second protocol, we tested these antagonists in 7 anesthetized, paralyzed, closed-chest dogs with acute left lower lobe atelectasis. Two manifestations of hypoxic vasoconstriction were examined: shunt fraction (as an inverse indicator of regional constriction in response to local hypoxia) and the pulmonary pressor response to global alveolar hypoxia (as an index of general hypoxic vasoconstriction). During normoxia before administration of the inhibitor, shunt fraction, measured using an SF6 infusion, was 25 +/- 7%. The pulmonary pressor response to hypoxia, defined as the increase in pulmonary end-diastolic gradient (PDG) produced by 10% O2 inhalation, averaged +10.5 +/- 3.6 mmHg. The increase in pulmonary vascular resistance (PVR) with hypoxia was +2.4 +/- 1.7 mmHg/L/min. Then, during normoxia, 1 of the 2 antagonists was administered. Shunt fraction was unchanged (26 +/- 4%; p = 0.5). The pressor response to hypoxia was slightly less but remained substantial (the increase in PDG with hypoxia was +7.9 +/- 2.8 mmHg; p less than 0.05; the increase in PVR was +1.8 +/- 1.2 mmHg/L/min, p less than 0.10). In the third protocol we gave RG 5901, a relatively specific 5 lipoxygenase inhibitor, to 5 dogs with lobar atelectasis. The indices of hypoxic vasoconstriction were not affected by RG 5901. Shunt fraction was 29.5 +/- 8.1% before and 27.0 +/- 7.4% after RG 5901 (p greater than 0.05). The pressor response to hypoxia was +8.9 +/- 2.1 mmHg before and +8.7 +/- 3.7 mmHg after RG 5901 (p greater than 0.05). We conclude that in dogs, hypoxic vasoconstriction does not appear to be mediated by leukotrienes. PMID- 2567792 TI - Trial of low-dose aspirin plus dipyridamole versus anticoagulants for prevention of aortocoronary vein graft occlusion. AB - In a prospective randomised trial, 249 patients who had aortocoronary vein bypass surgery were assigned either to a platelet inhibitory drug regimen or to standard anticoagulant therapy. Treatment was replaced by placebo in half of the patients in each group after 3 months. The platelet inhibitory drug regimen--very low-dose aspirin combined with dipyridamole--was as effective as standard anticoagulant therapy to prevent early and late graft occlusion. Death, myocardial infarction, and severe bleeding occurred significantly more often in patients receiving anticoagulants, whereas mild drug-related gastrointestinal and cerebral side effects were more common in patients taking platelet inhibitory drugs. Antithrombotic treatment should be continued for at least 1 year after coronary artery bypass graft surgery. PMID- 2567793 TI - Role of cholecystokinin in regulation of gastrointestinal motor functions. AB - By means of loxiglumide, a potent and highly specific antagonist for cholecystokinin (CCK), the effects of blocking CCK receptors on gastrointestinal motility were investigated in a placebo-controlled study in healthy young men (aged 21-39, mean 24 years). Gallbladder contraction stimulated by ingestion of a liquid test meal was completely abolished by oral administration of loxiglumide 30 min before the test meal. Gastric emptying of radio-opaque markers ingested with the test meal was significantly accelerated by loxiglumide (area under the curve [markers x h] 33.3 [SEM 3.8] vs 17.9 [2.7] after placebo). No effect of loxiglumide was found on small-bowel transit time, but 7 days' treatment with oral loxiglumide (800 mg three times daily) significantly shortened colonic transit time (29.4 [4.1] h after placebo, 15.0 [3.4] h after loxiglumide). It is concluded that CCK is an important mediator of meal-induced gallbladder contraction and is involved in the regulation of gastrointestinal motility in man. PMID- 2567795 TI - Transvaginal ultrasound. PMID- 2567794 TI - Diagnosis of Gerstmann-Straussler syndrome in familial dementia with prion protein gene analysis. AB - The polymerase chain reaction was used to screen DNA samples from 12 unrelated individuals with various familial dementias and ataxias for mutation in part of the prion protein (PrP) gene, an abnormality that occurs in individuals with the spongiform encephalopathies, Gerstmann-Straussler syndrome (GSS) and Creutzfeldt Jakob disease. 2 members of a family in whom GSS was not previously suspected had a 0.15 kb insertion of similar size to that found in another kindred with pathologically proven spongiform encephalopathy. GSS may be more common than is currently realised; PrP gene analysis is potentially useful for diagnosis and genetic counselling in familial dementias and ataxias. PMID- 2567796 TI - Polycythaemia due to hypoxaemia: advantage or disadvantage? PMID- 2567797 TI - Ultrasound/PEME for perineal pain. PMID- 2567799 TI - Laser and the ophthalmologist. PMID- 2567798 TI - Diagnosis of deep-vein thrombosis. PMID- 2567800 TI - In-situ breast cancer: the EORTC consensus meeting. PMID- 2567801 TI - Survival and causes of death in thalassaemia major. AB - Survival and causes of death were studied in 1087 Italian patients with thalassaemia major who were born on or after Jan 1, 1960. At the age of 15 years, the Kaplan-Meier estimate of survival after the first decade of life was 80.6% for subjects born in 1960-64, 84.2% for those born in 1965-69, and 96.9% for those born in 1970-74. At the age of 20 years, survival from the age of 10 was 59.1% for patients born in 1960-64, and 70.2% for those born in 1965-69; at 25 years, survival from the age of 10 was 40.7% in the 1960-64 cohort. Overall survival from birth for patients born in 1970-74 was 97.4% at 10 years, and 94.4% at 15 years. The most common cause of death was heart disease, followed by infection, liver disease, and malignancy. PMID- 2567803 TI - Health inequalities in Britain and Sweden. AB - Health differences between social classes are greater in Britain than in Sweden, but persist in both countries despite central Government interventions aimed at the reduction of such differences. PMID- 2567802 TI - Peptide regulatory factors and malignancy. PMID- 2567804 TI - Dermatofibromas and arthropod bites: is there any evidence to link the two? AB - 30 patients with dermatofibromas were questioned closely about contacts with stinging or blood-sucking arthropods before the development of their lesions. Their responses were not significantly different from those of a matched control group. 100 dermatofibromas were examined by dissection, histology, and after acid digestion of the collagenous material; no traces of arthropod skeletal tissues were found. These data are not consistent with the theory that the mechanical presence of arthropod tissues is necessarily, or even usually, a cause of dermatofibromas. PMID- 2567805 TI - Who needs steroid receptor assays? PMID- 2567806 TI - Cholesterol and mortality in elderly women. PMID- 2567807 TI - Lewis negative genotype and breast cancer risk. PMID- 2567808 TI - Effect of salt restriction on hypertension. PMID- 2567809 TI - Diabetes mellitus in Africans. PMID- 2567811 TI - Drainage of malignant effusions. PMID- 2567812 TI - Potential confusion of neuroleptic malignant syndrome and Wilson's disease. PMID- 2567810 TI - Side-effects of pamidronate. PMID- 2567813 TI - Atropine as possible contaminant of comfrey tea. PMID- 2567814 TI - Mobility of injection drug users and transmission of HIV. PMID- 2567815 TI - Decline in skeletal muscle mitochondrial respiration chain function with ageing. PMID- 2567816 TI - Fenoterol and fatal asthma. PMID- 2567817 TI - Jejunal delivery of levodopa methyl ester. PMID- 2567818 TI - Endothelin in urine. PMID- 2567819 TI - Panic and hypertension associated with single dose of buspirone. PMID- 2567820 TI - Radioiodine and aggravation of Graves' ophthalmopathy. PMID- 2567821 TI - Venous mesenteric ischaemia: conservative or surgical treatment? PMID- 2567822 TI - Mitral valve prolapse. PMID- 2567823 TI - Mitochondrial function in Parkinson's disease. PMID- 2567824 TI - Speech dysfluency. PMID- 2567825 TI - Academic boycott of South Africa. PMID- 2567826 TI - Home heating and renal failure. PMID- 2567827 TI - Evaluating gallbladder lithotripsy. PMID- 2567828 TI - International versus domestic health: an antiquated distinction. PMID- 2567829 TI - Beriberi secondary to tooth decay. PMID- 2567830 TI - Second trimester screening for trisomy 21. PMID- 2567831 TI - Sudden infant death and long-chain 3-hydroxyacyl-CoA dehydrogenase. PMID- 2567832 TI - Sex ratio and assisted conception. PMID- 2567833 TI - Transvaginal oocyte aspiration and pelvic infection. PMID- 2567834 TI - Increased plasma concentrations of endothelin-1 and big endothelin-1 in acute myocardial infarction. PMID- 2567835 TI - Passive smoking and low birthweight. PMID- 2567836 TI - Teniposide-induced hypersensitivity reactions in children. PMID- 2567837 TI - Informed consent and 25% risk of paralysis. PMID- 2567839 TI - Immunological treatment for multiple sclerosis. PMID- 2567838 TI - Disturbance of cerebral function by aluminium in haemodialysis patients without overt aluminium toxicity. AB - The psychomotor function of 27 long-term haemodialysis patients with apparently normal cerebral function, who had only mildly raised serum aluminium (mean 59 [SEM 9] micrograms/l), was measured by means of a computerised version of the symbol digit coding test. Compared with those of control subjects matched for age and the patients' estimated premorbid IQ, the patients' response times were significantly longer (2.51 [0.10] vs 1.88 [0.05] s). Abnormalities were also detected in five other computerised tests of psychomotor function. The mean activity of erythrocyte dihydropteridine reductase (DHPR), which is inhibited by aluminium, rose during 3 months' desferrioxamine treatment in most of the 15 patients so treated. Although there was no relation between baseline psychomotor function and either indices of cumulative aluminium exposure or erythrocyte DHPR activity, changes in DHPR induced by desferrioxamine correlated with changes in psychomotor performance (r = 0.62). The flash-stimulated visual evoked potential (measured in 10 patients) was delayed (133.4 [2.4] ms), although the pattern stimulated visual evoked potential remained normal (101.8 [3.2] ms). The difference between the visual evoked potentials stimulated by flash and pattern was significantly greater in the patients than in the controls (31.6 [4.3] vs 19.4 [2.4] ms) and was significantly related to the symbol digit coding response times and to the oral aluminium intake. The results suggest that much more rigorous exclusion of aluminium from the dialysate and diet of dialysis patients is necessary. PMID- 2567840 TI - Hurricane Gilbert anaemia. PMID- 2567841 TI - Genetic differences between cystic fibrosis with and without meconium ileus. PMID- 2567842 TI - Percutaneous excimer laser coronary angioplasty. PMID- 2567843 TI - Long-term effects of methyl isocyanate. PMID- 2567844 TI - Acute ifosfamide-induced tubular toxicity. PMID- 2567845 TI - Limited value of chest radiography in uncomplicated hypertension. PMID- 2567846 TI - Interaction of fluconazole with cyclosporin. PMID- 2567847 TI - Cervical columnar epithelium and tylosis. PMID- 2567848 TI - Psychosis and anti-tuberculosis therapy. PMID- 2567849 TI - Cuba's health record. PMID- 2567851 TI - Tobacco and the Karolinska. PMID- 2567850 TI - The Oregon Health Initiative. PMID- 2567852 TI - Dipyrone trials in Thailand. PMID- 2567854 TI - Pentamidine salts. PMID- 2567853 TI - Gangrenous stomatitis in AIDS. PMID- 2567855 TI - Reactivation of hepatitis B in AIDS. PMID- 2567856 TI - Medical technology in Italy. PMID- 2567857 TI - Premarital sex in rural India. PMID- 2567858 TI - Vesicovaginal fistula. PMID- 2567859 TI - Use of ghee during delivery. PMID- 2567860 TI - Epstein-Barr virus/complement receptor and epithelial cells. PMID- 2567861 TI - Dietary cholesterol and heart disease. PMID- 2567862 TI - Potential toxic interactions of cocaine and mazindol. PMID- 2567863 TI - Classification of hypertensive disorders of pregnancy. PMID- 2567864 TI - Salmonella infection after ileoanal anastomosis. PMID- 2567865 TI - Indications for flecainide. PMID- 2567867 TI - Extracorporeal shockwave lithotripsy of salivary gland stones. PMID- 2567866 TI - Cigarettes and heart disease. PMID- 2567868 TI - Blood cholesterol measurement in the prevention of coronary heart disease. PMID- 2567869 TI - Duodenal bulb acidity and the natural history of duodenal ulceration. AB - Fasting and post-prandial duodenal bulb pH was recorded in 10 subjects with untreated duodenal ulcers (DU), 10 subjects with healed DU, and 15 healthy volunteers. During fasting, pH was less than 4 for a greater percentage of time in healed than in active DU. Ingestion of a solid test meal accentuated this difference: subjects with active DU and controls both showed less bulb acidification than subjects with healed ulcers. Bulb pH was similar in patients with active ulceration and controls. The finding of neutral bulb pH with active ulceration and low bulb pH with healing may explain the typical cycle of spontaneous healing and relapse in duodenal ulcer disease. PMID- 2567870 TI - Cytomegalovirus infection and progression towards AIDS in haemophiliacs with human immunodeficiency virus infection. AB - To examine whether cytomegalovirus (CMV) infection could accelerate progression of human immunodeficiency virus (HIV) infection to AIDS, serological studies were done on 108 HIV-infected haemophiliacs. In the 1.3-9 years from time of first recognised HIV seroconversion, the age-adjusted risk of CDC group IV disease in CMV-seropositive patients was 2.5 times that in CMV-seronegative patients. CMV seropositive patients were also more likely to have detectable p24 antigenaemia. Survival analysis showed that CMV-seropositive patients were at greater risk of HIV disease than CMV-seronegative patients from about 2 years after HIV seroconversion. Thus CMV infection is associated with a more rapid progression to HIV disease. PMID- 2567871 TI - Galactose consumption and metabolism in relation to the risk of ovarian cancer. AB - In a case-control study, consumption of dairy foods by 235 white women with epithelial ovarian cancer and by 239 control women, and activity of red blood cell galactose-1-phosphate uridyl transferase (transferase) in a subset of 145 cases and 127 controls were determined. Yogurt was consumed at least monthly by 49% of cases and 36% of controls. The mean transferase activity of cases was significantly lower than that of controls. When a ratio of lactose consumption to transferase (L/T) was calculated, cases had a mean L/T of 1.17 compared with 0.98 for controls; there was a highly significant trend for increasing ovarian cancer risk with increasing L/T ratio. Lactose consumption may be a dietary risk factor and transferase a genetic risk factor for ovarian cancer. PMID- 2567872 TI - Predominance of Borrelia burgdorferi specific B cells in cerebrospinal fluid in neuroborreliosis. AB - A nitrocellulose immunospot assay that allows the counting of cells secreting IgG, IgA, or IgM antibodies to Borrelia burgdorferi was used to compare B cell response to B burgdorferi at the cellular level in cerebrospinal fluid (CSF) and blood from patients with neuroborreliosis with that in patients with aseptic meningoencephalitis (AM) or non-inflammatory neurological diseases. 13 of the 14 patients with untreated neuroborreliosis had CSF cells secreting IgG antibodies to B burgdorferi (mean 17 cells per 10(4) CSF cells), whereas 8 of 12 patients examined had cells secreting IgA antibodies (mean 6 cells) and 10 of 12 had cells secreting IgM antibodies (mean 6 cells) per 10(4) CSF cells. IgG antibody producing cells predominated except in 2 patients with mainly or only IgM secreting cells. Cells secreting antibodies to B burgdorferi were rarely found in the blood and then at very low numbers, which reflects preferential compartmentalisation of the specific B cell response to the CSF. The cells were not detectable in CSF or blood from the two control groups. Evaluation of humoral immunity at the cellular level is a novel approach to the detection and localisation of immune events in neuroinflammatory disorders. PMID- 2567873 TI - Randomised, controlled trial of squatting in the second stage of labour. AB - A new obstetric aid, the 'Birth Cushion' allows the parturient to sink into a supported squatting posture for the second stage of labour and delivery; it fits onto conventional delivery beds. A prospective, controlled trial of 427 primiparae compared the outcome of labour in women randomly allocated to squatting (218) or conventional semirecumbent (209) management. The squatting group had significantly fewer forceps deliveries (9% vs 16%) and significantly shorter second stages (median length of pushing 31 vs 45 min) than the semirecumbent group. There were fewer perineal tears, but more labial tears, in the squatting group. Apgar scores, blood loss, and post-partum vulvar oedema were similar in both groups. 82% of the women in the squatting group maintained upright positions for most of the second stage, and reported great satisfaction with the supported squatting position. The traditional birth posture of squatting can be easily adapted for modern labour management and has advantages for women in their first labour. PMID- 2567875 TI - Irish electorate speaks on health. PMID- 2567874 TI - Digoxin: new answers; new questions. PMID- 2567876 TI - Photodynamic therapy. PMID- 2567878 TI - Anyone for IFDO? PMID- 2567877 TI - Lupus nephritis and pregnancy. PMID- 2567879 TI - Incidence and sonographic diagnosis of bacterial ileocaecitis masquerading as appendicitis. AB - 533 consecutive patients with suspected acute appendicitis or appendiceal mass were examined by ultrasonography to distinguish acute appendicitis from bacterial enteritis. In 61 (11.4%) ultrasonography revealed the characteristic picture associated with bacterial enteritis of the ileocaecal region--enlarged mesenteric lymph nodes and mural thickening of the terminal ileum and caecum--but no image of the appendix. In 41 of these a bacterial infection was confirmed: infection due to Yersinia enterocolitica in 21, Campylobacter jejuni in 15, Salmonella B in 3, Salmonella C in 1, and Yersinia pseudotuberculosis in 1. In the other 20 bacteriological tests were negative (10) or not done (10). Oral barium studies, done in 15 patients, showed thickening of the terminal ileum in all of them. Only 22 of the 61 patients had diarrhoea. Yersinia enteritis clinically simulated an appendiceal mass in 17 of 22 patients, 6 of the 61 patients underwent surgery, and in all of them the appendix removed was normal. The other 55 patients recovered with conservative treatment. In 26 a planned appendicectomy was cancelled because of the sonographic findings. Bacterial enteritis limited to the ileocaecal region (bacterial ileocaecitis) seems to be responsible for an appreciable number of unnecessary appendicetomies. It has characteristic sonographic features that distinguish it from appendicitis. PMID- 2567880 TI - Contamination of endoscopes used in AIDS patients. AB - Contamination of twenty endoscopes used in patients with AIDS was assessed. The suction-biopsy, air, and water channels and the insertion tube were sampled after use, after washing in detergent, and after disinfection for 2 min in 2% alkaline glutaraldehyde. The polymerase chain reaction with Southern blotting, cell cultures, and antigen immunoassay were used to detect human immunodeficiency virus (HIV). Samples were also examined for cytomegalovirus, adenoviruses, enteroviruses, herpes simplex virus, myxoviruses, hepatitis B surface antigen, fungi, and bacteria. Seven of twenty unwashed endoscopes were contaminated by HIV. Commensal bacteria were found in all endoscopes, Candida albicans in six, Staphylococcus aureus in five, and Pseudomonas aeruginosa in five. Washing alone removed all detectable organisms from 66 of 68 contaminated sites; Neisseria spp were recovered from two air channels after washing but not after disinfection. Washing achieved a mean reduction of 4.93 (95% confidence interval 3.69-6.17) colony forming units per ml. PMID- 2567881 TI - Indicators of effective cytological sampling of the uterine cervix. AB - A case-control comparison of adequate and inadequate cervical smears (adequacy being defined as the successful detection of cervical intraepithelial neoplasia and inadequacy as the failure to do so when disease was almost certainly present) was undertaken to determine whether indicators of effective cytological sampling could be identified. The association between inadequate smears and the absence of two types of normal epithelial cells (columnar cells of endocervical origin and immature metaplastic cells) was measured. A significant and substantial association was found between inadequate cervical smears and immature metaplastic cells and between inadequate smears and both types of cell. Endocervical cells alone were less likely to be found in inadequate than in adequate smears, but this association was not statistically significant. PMID- 2567882 TI - Zidovudine-induced neutropenia: are we too cautious? AB - 30 patients with recurrent zidovudine-induced neutropenia were followed up for a total of 493 months of treatment to evaluate their risk of bacterial infection. Zidovudine was temporarily discontinued only when the polymorphonuclear (PMN) cell count fell to less than 500/microliters. The incidence of bacterial infection during periods of severe neutropenia (PMN less than 500/microliters) was 230% higher than when the PMN count was 500-1000/microliters, and 600% higher than when the count was greater than 1000/microliters. The difference between periods when the PMU count was 500-1000/microliters and non-neutropenic periods was not significant. The findings suggest that zidovudine therapy can be continued despite neutropenia without a major increase in the incidence of bacterial infection provided the PMN count does not fall to less than 500/microliters. PMID- 2567883 TI - Drugs in donated blood. AB - Blood from donors who are or who have been on drug treatment may be hazardous to the recipient. Guidelines are proposed to increase the safety of donated blood without needless rejection of donors, based on the time that should elapse between the last dose and safe blood donation for a range of commonly used drugs. PMID- 2567884 TI - Medicine for sale. PMID- 2567885 TI - ACE inhibitors in pregnancy. PMID- 2567886 TI - Treatment of chronic proctosigmoiditis with cyclosporin enemas. PMID- 2567887 TI - Oral contraceptives and breast cancer. PMID- 2567888 TI - Mumps meningitis following measles, mumps, and rubella immunisation. PMID- 2567889 TI - Screening for bacteriuria in elderly people. PMID- 2567890 TI - Percutaneous angioscopy as adjunct to laser angioplasty in peripheral arteries. PMID- 2567891 TI - Radon exposure and leukaemia. PMID- 2567892 TI - Cerebral perfusion deficits in dysbaric illness. AB - Decompression sickness (DCS) is usually categorised as type I (mild; peripheral pain, non-neurological) or type II (serious; neurological). Type II is regarded as predominantly a spinal cord disease with infrequent cerebral involvement. Cerebral perfusion was studied by injection of 99Tcm-hexamethylpropyleneamine oxime and single photon emission tomography in 28 divers with confirmed incidents of DCS and cerebral arterial gas embolism (CAGE). Cerebral perfusion deficits were present in all 23 cases of type II DCS and in all 4 cases of CAGE. No deficits were present in the single case of type I DCS. Type II DCS should be recognised as a diffuse, multifocal, central nervous system disease. PMID- 2567893 TI - Tranexamic acid for control of haemorrhage in acute promyelocytic leukaemia. AB - In a double-blind study, 12 consecutive patients with acute promyelocytic leukaemia were randomised either to tranexamic acid (TA group) or to placebo (control group) for 6 days to see whether inhibition of fibrinolysis would reduce haemorrhage and transfusion requirements. The total study period was 14 days. In the TA group, there were fewer haemorrhagic episodes, as determined by a scoring system. Packed red cell transfusion requirements decreased; and fewer additional platelet concentrate transfusions were needed. These beneficial effects were more pronounced in the second week. There were no thromboembolic complications. PMID- 2567894 TI - Inositol 1,4,5-trisphosphate phosphatase deficiency and malignant hyperpyrexia in swine. AB - The sarcoplasmic reticulum from muscle of swine which are susceptible to malignant hyperpyrexia is deficient in inositol 1,4,5-trisphosphate phosphatase (InsP35-ase) activity, which leads to high intracellular concentrations of inositol 1,4,5-trisphosphate (InsP3) and of calcium ions. Halothane inhibits InsP35-ase and further increases myoplasmic InsP3 and calcium ion concentrations, and produces the clinical features of malignant hyperpyrexia. PMID- 2567895 TI - Idiopathic sclerosing encapsulating peritonitis. AB - Sclerosing encapsulating peritonitis (SEP) was diagnosed in two men, one middle aged woman, and a prepubertal girl, none of whom had been treated with beta blockers, peritoneal dialysis, or peritoneal shunting. Idiopathic SEP has hitherto been reported mainly in adolescent females. It is hypothesised that in these patients, who were from two adjacent districts of Rajasthan, an aetiological factor was consumption of cereals infected with fungus that contained biologically active amines. PMID- 2567896 TI - Hyperglycaemia and absorption of sulphonylurea drugs. AB - To examine whether hyperglycaemia impairs the absorption of sulphonylurea agents, glipizide, which is rapidly and completely absorbed, was measured in plasma from 12 healthy young subjects during various levels of experimentally-induced hyperglycaemia. An increase in the plasma glucose concentration above 7 mmol/l was associated with a dose-dependent delay in the absorption of glipizide; at a concentration above 11 mmol/l, the plasma glipizide concentration was reduced by 50%. The data indicate that hyperglycaemia may delay the absorption of sulphonylurea agents, probably because it impairs gastric motility and/or gastric emptying. This delay of absorption may be clinically relevant, since the efficacy of short-acting sulphonylureas is dependent upon the absorption rate of the drug. PMID- 2567898 TI - Neonatal seizures. PMID- 2567897 TI - Effect of perioperative blood transfusion on recurrence of Crohn's disease. AB - The effect of perioperative blood transfusion on recurrence of Crohn's disease was investigated retrospectively in 60 patients with Crohn's disease who underwent small-bowel resection. 28 patients received 1-8 units of blood; the others were not transfused. Both groups were similar in age, duration of Crohn's disease, and preoperative serum albumin, but preoperative haemoglobin, length of intestine resected, and location of small-bowel involvement were different. The patients who received perioperative blood transfusion had a significantly lower recurrence rate by life-table analysis, although the site and greater length of bowel involvement in the transfused patients would normally indicate an increased likelihood of recurrence. 5 years after bowel resection, the cumulative recurrence rate in transfused patients was 19%, compared with 59% in controls. The immunosuppressive effect of blood transfusion may modify the progression of Crohn's disease. PMID- 2567899 TI - Psychiatric intervention after disaster. PMID- 2567900 TI - Mitral valve and wedge pressure. PMID- 2567901 TI - Measuring therapeutic risk. PMID- 2567903 TI - Randomised study of myringotomy, amoxycillin/clavulanate, or both for acute otitis media in infants. AB - In a prospective study, 105 infants aged 3-12 months with acute otitis media were randomly assigned to one of three treatment groups: amoxycillin/clavulanate ('Augmentin') alone (36 patients), myringotomy plus placebo (35 patients), or augmentin plus myringotomy (34 patients). The last two groups were double blinded. Bacterial pathogens, mainly Haemophilus influenzae (of which 20% were beta-lactamase producers) and Streptococcus pneumoniae, were isolated from 60% of the ear exudates and all isolates were sensitive to augmentin. Most of the infants improved clinically within 3-6 days irrespective of the treatment protocol. As judged by otoscopy, 60% of the patients receiving augmentin, with or without myringotomy, recovered completely compared with 23% of patients treated with myringotomy plus placebo. Treatment with augmentin was also more effective than myringotomy with regard to persistence of ear infection. In the myringotomy plus augmentin group closure of the incision and resolution of the discharge from the incision site was faster than in the myringotomy plus placebo group. The addition of myringotomy to augmentin did not seem to affect either the persistence of the infection after treatment or the residual middle ear effusion. PMID- 2567902 TI - Lessons from history--maternal and infant mortality. PMID- 2567904 TI - Individual outcome prediction models for intensive care units. AB - Prognostic criteria based on static analysis of group statistics do not help much in decisions to withhold or withdraw therapy from intensive care unit (ICU) patients too ill to benefit, since they do not provide adequate information on the features that distinguish non-survivors from survivors. A predictive model which uses dynamic analysis of severity scores based on physiological variables is presented here along with the results of tests of the model in 831 ICU patients. 109 patients were correctly predicted to die by the model. Of 722 whose prediction was outcome unknown, 181 died. Thus, the odds for prediction of death among non-survivors were 0.376. Since there were no false predictions of death, the estimated chance of a false prediction was 0.0055. PMID- 2567905 TI - Post-exposure prophylaxis for hepatitis B: active or passive? AB - After exposure to hepatitis B (HB) virus, passive immunisation with HB immune globulin is widely used for protection while active immunity is induced by conventional vaccination regimens. Protective antibody titres can be achieved much more quickly with accelerated vaccination, and the role of passive immunisation may need to be reconsidered. PMID- 2567906 TI - Why not base clinical education in general practice? PMID- 2567907 TI - Thoughts and ethics. The place for thought in medical ethical decision-making. PMID- 2567908 TI - New non-operative treatment for variceal haemorrhage. PMID- 2567909 TI - Thrombolytic therapy. PMID- 2567910 TI - Screening for gestational glucose intolerance. PMID- 2567911 TI - Protease antigens from house dust mite. PMID- 2567912 TI - "Fingertip unit" in dermatology. PMID- 2567913 TI - Lactational hypomagnesaemia. PMID- 2567914 TI - Animal toxicology of iron chelator L1. PMID- 2567915 TI - 'Enterotest' and Strongyloides stercoralis. PMID- 2567916 TI - Echo-planar magnetic resonance imaging in abnormal pregnancies. PMID- 2567917 TI - Alpha-blockade for hypertension. PMID- 2567918 TI - Oral contraceptives and breast cancer. PMID- 2567920 TI - Comparison of latex agglutination with established bacteriological tests for diagnosis of cerebrospinal meningitis. PMID- 2567919 TI - Oral contraceptives, tobacco smoking, and breast cancer risk. PMID- 2567921 TI - Nephrotoxicity of ifosfamide in children. PMID- 2567922 TI - Naloxone hazard in infant of opioid abuser. PMID- 2567923 TI - Recombinant human erythropoietin treatment in patients on maintenance home haemodialysis. PMID- 2567924 TI - Preventing transfusion-acquired cytomegalovirus infection in infants. PMID- 2567925 TI - Carboplatin or cisplatin? PMID- 2567927 TI - Feminism in publishing. PMID- 2567926 TI - Chloramphenicol treatment of childhood bacterial meningitis. PMID- 2567928 TI - Obstetric protest. PMID- 2567929 TI - Traditional birth attendants. PMID- 2567930 TI - Cervical cytology policy. PMID- 2567931 TI - Heart disease in Tibet. PMID- 2567932 TI - Low cholesterol and increased risk. PMID- 2567934 TI - In-vivo anti-CD3 treatment of autoimmune patients. PMID- 2567933 TI - Tacrine in Alzheimer's disease. PMID- 2567935 TI - Echovirus 6 encephalitis in a preterm baby. PMID- 2567936 TI - Ceftriaxone-associated biliary pseudolithiasis in adults. PMID- 2567937 TI - Cocaine abuse and opioid withdrawal. PMID- 2567938 TI - HIV incubation times. PMID- 2567939 TI - Outbreak of HIV seropositivity among commercial plasma donors in Pune, India. PMID- 2567940 TI - Resurgence of heterosexually acquired early syphilis in London. PMID- 2567941 TI - Ovulation induction and neural tube defects. PMID- 2567942 TI - Treatment of biliary colic with loxiglumide. PMID- 2567943 TI - Complication of triptorelin treatment for uterine myomas. PMID- 2567944 TI - Donor kidneys of infants and very young children are unacceptable for transplantation. PMID- 2567945 TI - Conservation therapy of breast cancer. PMID- 2567946 TI - Congenital hypothyroidism and monoamniotic twins. PMID- 2567947 TI - Human papillomavirus subtype 16a. PMID- 2567948 TI - Scurvy and anaemia in refugees. PMID- 2567949 TI - UVA filters in sunscreen preparations. PMID- 2567950 TI - Plasma exchange and the anticardiolipin syndrome in pregnancy. PMID- 2567951 TI - Suxamethonium anaphylaxis. PMID- 2567952 TI - Hernia and hydrocele in cryptorchidism. PMID- 2567953 TI - Reactivation of hepatitis B. PMID- 2567954 TI - Total retroperfusion of the coronary arteries and veins. PMID- 2567955 TI - Right to know in Japan. PMID- 2567956 TI - Role of ovarian steroids on the catecholamine synthesis and release in female rat adrenal: in vivo and in vitro studies. AB - In a previous report, we describe the existence of an effect of ovarian steroids on the adrenal medulla activities of the enzymes involved in catecholamine (CA) catabolism. To complete that study, we have now examined the adrenal medulla activity of tyrosine hydroxylase (TH), the rate limiting enzyme of the CA synthesis, as well as the in vitro release of CAs from incubated adrenal medullas. The study has been performed with adrenal medullas from female rats with physiological (estrous cycle) or pharmacological (steroid treatment) alterations in their circulating levels of estrogens and progesterone. The in vitro release of CAs from incubated adrenal medullas of estradiol-treated rats was lower than that obtained in vehicle-treated animals. In consequence, the preovulatory increase of estradiol would be the responsible of the low in vitro release of CAs observed during the estrous phase of ovarian cycle. However, this steroid does not seem to affect the CA synthesis, since the adrenal medulla activity of TH was not altered after the estradiol treatment nor during the estrous cycle. On the contrary, progesterone treatment increased TH activity 24 h after the steroid injection. This effect was independent of estradiol. However, an estrogen-dependent increase in TH activity occurred short-time after the steroid administration. Although progesterone by itself failed to modify the in vitro release of both CAs, it was able to reverse the estradiol-induced decrease in epinephrine release. In summary, estradiol seems to decrease the ability of the adrenal medulla to release CAs to the peripheral blood, without affecting the CA synthesis, whereas progesterone mostly affects TH activity, being its effects temporary and partially depending on estrogens. PMID- 2567958 TI - Experimental diabetes increases the formation of sulfane by transsulfuration and inactivation of tyrosine aminotransferase in cytosols from rat liver. AB - The addition of L-cysteine to hepatic cytosols causes inactivation of tyrosine aminotransferase. We have studied the mechanism of inactivation and the effect of streptozotocin-induced diabetes in the rat on the inactivation of tyrosine aminotransferase in the presence of fractions prepared from livers and kidneys. Diabetes increased the rate at which tyrosine aminotransferase was inactivated after addition of cysteine to hepatic cytosols. The inactivation was due to the production of thiocysteine (which contains sulfane sulfur) from cystine as a result of desulfuration catalyzed by gamma-cystathionase. Diabetes increased the content of cystathionine beta-synthase and gamma-cystathionase in liver. As a result, cytosols from diabetic animals converted homocysteine, cystathionine, cysteine and cystine to sulfane at an elevated rate, with resulting inactivation of tyrosine aminotransferase. In contrast, inactivation in kidney fractions was not affected by diabetes. Incubation with an inhibitor of gamma-cystathionase (propargylglycine) prevented inactivation of tyrosine aminotransferase. These results show that the potential for the formation of sulfane sulfur by the enzymes of the transsulfuration pathway is enhanced by chronic diabetes. PMID- 2567957 TI - [Nizatidine]. AB - Nizatidine is a new H2-receptor antagonist, as potent as ranitidine. It does not interfere with hepatic metabolism and it lacks anti-androgenic effects. An evening dose of 300 mg suppresses nocturnal acid secretion without diurnal carryover. Published clinical trials are limited. Over 3800 patients who have been treated with nizatidine, 300 mg for up to eight weeks or 150 mg at night for a year, did not appear to have significantly unexpected or unwanted side effects. Rate of healing for both duodenal and gastric ulcers and reduction in the rate of relapse are comparable to those of ranitidine. PMID- 2567959 TI - Decline of hemopoietic stem cells during serial transplants in lethally irradiated recipients. AB - Stochastic and field models of hemopoietic stem cell renewal are tested for quantitative predictions regarding limited survival of mouse stem cells during serial transplant from donors into lethally irradiated recipients. Besides circumventing the limitations of stochastic theories, the field model is found to be congruent with experimental data. PMID- 2567960 TI - Octreotide--a synthetic somatostatin. PMID- 2567961 TI - The presence of amplified regions affects the stability of chromosomes in drug resistant Chinese hamster cells. AB - The stability of chromosomes carrying amplified CAD (carbamyl phosphate synthetase-aspartate transcarbamylase-dihydroorotase) or DHFR (dihydrofolate reductase) genes was studied in V79 Chinese hamster cell derivatives resistant to PALA (N-phosphonacetyl-L-aspartate) and MTX (methotrexate), respectively. Cells were maintained in the presence of the selective drugs during the study. In both metaphase chromosomes and interphase nuclei, amplified regions were localized by in situ hybridization. In MTX-resistant cells, the amplification-bearing chromosome moved sluggishly at anaphase and gave rise to bud-shaped formations in interphase nuclei. It is suggested that these buds could eventually separate as micronuclei. In both MTX- and PALA-resistant cells, amplified DNA was observed in micronuclei in interphase and in displaced chromosomes in metaphase. Finally, amplification-bearing dicentric chromosomes were found in both drug-resistant cell lines. Cumulatively, these observations indicate that the presence of the amplified region in a chromosome renders it unstable: chromosomes bearing an amplified region tended to be excluded from cells, and rearrangements were more frequent than in normal chromosomes. PMID- 2567962 TI - Expression of dopaminergic phenotypes in the mouse olfactory bulb induced by the calcitonin gene-related peptide. AB - In the olfactory bulb, tyrosine hydroxylase (TH), the rate-limiting enzyme in the biosynthesis of catecholamines, is expressed after birth when the axons of olfactory epithelial neurons have made synapses in the bulb. It has been suggested that expression of TH is regulated trans-synaptically because on deafferentation of the bulb there is a marked decrease in the contents of TH, dopamine and 3,4-dihydroxyphenylacetic acid, which, however, return to normal levels after regeneration of the primary afferents. To date the molecular signalling involved in this trans-synaptic induction has not yet been characterized; I have therefore studied the expression of dopaminergic properties (presence of TH and dopamine uptake) in dissociated cell cultures from embryonic mouse olfactory bulb. I report that the number of dopaminergic cells increases fivefold when olfactory bulb neurons are co-cultured with olfactory epithelial neurons and that soluble factors, rather than cell interactions, mediate this effect. The dopaminergic-inducing factor is the calcitonin gene-related peptide (CGRP) which is present in chemosensory neurons of the olfactory epithelium and when added at nanomolar concentrations to olfactory bulb cultures mimics the effect of olfactory epithelial neurons. Significantly the induction of dopaminergic phenotypes brought about by olfactory epithelial neurons is abolished by an antiserum to CGRP. These observations show that CGRP is involved in the differentiation of dopaminergic olfactory bulb neurons. PMID- 2567964 TI - Mitogenic neurotransmitters. PMID- 2567963 TI - Neurotransmitter inhibition of neuronal calcium currents by changes in channel voltage dependence. AB - The voltage-dependent calcium current of many neurons is depressed by transmitters such as noradrenaline, GABA, and kappa-opiate agonists. This modulation probably constitutes a major mechanism of presynaptic inhibition. Although recent work has implicated GTP-binding proteins in the mechanism of current inhibition, it is still unknown how the activation of those proteins alters the operation of the channels. In their initial description of the phenomenon, Dunlap and Fischbach proposed that noradrenaline acts by somehow reducing the number of functions calcium channels in the cell. By contrast with this hypothesis, I have found that inhibition of Ca2+ current is primarily due to a transmitter-induced change in the voltage-dependence with which channels are opened. Transmitters profoundly alter the voltage-dependence of channel activation, but there is little or no change in the number of functional channels activated by very large depolarizations. There is also little effect on the voltage-dependence of inactivation. PMID- 2567965 TI - Effect of the D1 receptor agonist SKF 38393 on some behavioural effects of apomorphine in rats. AB - The dopamine receptor agonist apomorphine in experiments on rats in low doses (0.025-0.2 mg/kg, s.c.) induced yawning which reflected a selective activation of presynaptic dopamine receptors. In high doses (0.25-1.0 mg/kg) apomorphine induced stereotyped sniffing and yawning in consequence of postsynaptic D2 receptor activation. Dopamine D1 receptor agonist SKF 38393 inhibited yawning induced by low doses of apomorphine. The inhibitory effect of SKF 38393 on apomorphine-induced yawning was attenuated by pretreatment with specific D1 receptor antagonist SCH 23390 [2-(+)-8-chloro-2,3,4,5-tetrahydro-3-methyl-5 phenyl-1H-3-benzazepine-7-ol). On the other hand however, SKF 38393 potentiated sniffing induced by the high doses of apomorphine without affecting gnawing. These data indicate that D1 receptor activation modulates both pre- and postsynaptic effects of apomorphine in opposite directions. PMID- 2567966 TI - Reflex epicardial coronary vasoconstriction elicited by nicotine in anaesthetized dogs. AB - The effects of arterial chemoreceptor activation by nicotine on coronary artery diameter was studied in anaesthetized, artificially ventilated dogs. Left circumflex coronary artery diameter, coronary blood flow, calculated mean coronary resistance, systemic arterial blood pressure and heart rate were measured. In control dogs (n = 10) the injection of nicotine (100 micrograms) into the carotid artery evoked an increase of arterial pressure (+22 +/- 9 mm Hg) and a decrease in heart rate (-36 +/- 13 beats/min), and tended to increase coronary blood flow (+7 +/- 4 ml/min). Intracarotid nicotine had no effect on large coronary artery diameter (+0.02 +/- 0.03 mm) or total coronary resistance (+0.04 +/- 0.09 mm Hg min/ml) under these conditions. When heart rate was controlled by (1) beta-adrenoceptor blockade (propranolol, 1 mg/kg i.v.) plus pacing of the right ventricle (n = 4) or (2) beta-adrenoceptor blockade plus bilateral vagotomy (n = 7), the chemoreflex-induced constriction of the large coronary artery (-0.07 +/- 0.02 mm and -0.12 +/- 0.03 mm, respectively; p less than 0.05). In contrast, there was no chemoreflex-induced change in total coronary resistance after beta-adrenoceptor blockade plus pacing (+0.01 +/- 0.09 mm Hg min/ml, but after beta-adrenoceptor blockade plus vagotomy coronary resistance was increased (+0.75 +/- 0.31 mm Hg min/ml; p less than 0.05). The constriction of both large and small coronary arteries was abolished by phentolamine (0.5 mg/kg i.v.). These results suggest that carotid body chemoreceptor stimulation by nicotine can produce reflex alpha-adrenoceptor mediated constriction of both large and small coronary arteries, and that the constriction of the small vessels is balanced by vagally-mediated dilatation. PMID- 2567967 TI - Subtype-selective up-regulation of human saphenous vein beta 2-adrenoceptors by chronic beta-adrenoceptor antagonist treatment. AB - To study beta-adrenoceptor antagonist-induced changes in human vascular beta adrenoceptors, we determined the effects of chronic treatment with different beta adrenoceptor antagonists without intrinsic sympathomimetic activity (ISA) on beta 2-adrenoceptor density (assessed by (-)-[125I]-iodopindolol binding) in human saphenous vein membranes obtained from patients undergoing coronary artery bypass grafting. In patients chronically treated with the non-selective beta adrenoceptor antagonists propranolol or sotalol, the density of saphenous vein beta 2-adrenoceptors was significantly higher than in control (i.e. patients not treated with beta-adrenoceptor antagonists), whereas in patients chronically treated with the selective beta 1-adrenoceptor antagonists metoprolol or bisoprolol it was not different from control. It is concluded that beta adrenoceptor antagonists without ISA increase human saphenous vein beta 2 adrenoceptors in a subtype-selective fashion. PMID- 2567969 TI - [Antihypertensive agents: adrenergic beta receptor blockaders]. PMID- 2567968 TI - Dopamine inhibits prostaglandin F2 alpha-induced depolarization of rabbit jejunal arteries via activation of DA1-receptors. AB - In rabbit jejunal arteries, the membrane potential of single smooth muscle cells decreased on the application of noradrenaline 3 mumol/l. LY 171555 1 mumol/l did not change, whereas SKF 38393 10 mumol/l reversed the effect of noradrenaline. When prostaglandin F2 alpha (PGF2 alpha) was used to evoke depolarization in the presence of prazosin 0.1 mumol/l, rauwolscine 1 mumol/l and propranolol 1 mumol/l, both SKF 38393 10 mumol/l and dopamine 10 mumol/l repolarized the membrane. SCH 23390 1 mumol/l antagonized the effects of SKF 38393 10 mumol/l and dopamine 10 mumol/l. Thus, the change in membrane potential is mediated by a DA1 receptor. PMID- 2567970 TI - Polyarteritis nodosa and necrotizing glomerulonephritis associated with long standing silicosis. AB - A 55-year-old white male, with silicosis diagnosed 10 years earlier, presented massive proteinuria with microscopic hematuria, moderate renal failure and distal polyneuropathy. Bilateral renal angiography showed multiple intraparenchymal saccular aneurysms. Renal biopsy disclosed a focal segmental necrotizing glomerulonephritis and arteriolitis. After combined corticosteroid and immunosuppressive treatment, renal function improved and remained stable 6 months later. PMID- 2567971 TI - Apomorphine affects cholecystokinin content via preferentially D1 or D2 dopamine receptor according to the regions of the rat brain. AB - Cholecystokinin octapeptide-like immunoreactivity (CCK-8IR) was measured in several regions of the rat brain after the intraperitoneal administration of apomorphine, SKF-38393 (D1 agonist), LY-171555 (D2 agonist). In the medial prefrontal cortex and striatum, apomorphine and SKF 3839 decreased CCK-8IR. In the anterior and posterior nucleus accumbens, on the other hand, the inhibitory effect of apomorphine was mimicked by LY-171555. These results suggest that apomorphine affects CCK-8IR via either the D1 dopamine (DA)-receptor or D2 DA receptor according to the brain region. PMID- 2567972 TI - Effects of angiotensin II on cultured, bovine adrenal medullary cells. AB - Primary cultures of bovine adrenal medullary cells have been used to study the effects of angiotensin II on catecholamine secretion and inositol phosphate accumulation. Angiotensin II induced a weak secretion of both adrenaline and noradrenaline, with a threshold of 10-100 pM and a shallow concentration dependence up to 10 microM. The response was fully dependent on extracellular Ca++, was partially inhibited by 100 nM nifedipine, was completely blocked by [Sar1, Ala8]-angiotensin II (IC50 5-10 nM) and was unaffected by 0.1 mM hexamethonium. Angiotensin II also increased inositol phosphate accumulation over the range 1 pM-10 microM. Inositol trisphosphate levels increased in a biphasic manner after 15 sec and 1 min exposure to 10 nM angiotensin II, but were not significantly increased at 30 sec or 5, 15 or 30 min stimulation. Inositol bisphosphate was significantly increased after 1 min. Inositol monophosphate levels only increased after 1 min stimulation, but continued to rise during 30 min stimulation. Removal of extracellular Ca++ or addition of EGTA reduced basal inositol phosphate accumulation but not the ability of angiotensin II to stimulate inositol phosphate accumulation relative to basal. Nifedipine (100 nM) had no effect on basal or angiotensin II-induced inositol phosphate accumulation. The inositol phosphate response to angiotensin II was abolished by 1 microM [Sar1, Ala8]-angiotensin II. The results suggest that secretion of adrenal medullary catecholamines can be evoked by angiotensin II, at concentrations that are compatible with a role for circulating angiotensin II or for angiotensin II generated locally within the adrenal medulla. They do not support the suggestion that the secretory actions of angiotensin II on chromaffin cells are mediated by mobilization of intracellular Ca++ stores. PMID- 2567973 TI - Enhancement of human lymphocyte natural killer activity by somatostatin. AB - The effect of somatostatin 1-14 (SS) on the natural killer (NK) activity of human peripheral blood lymphocytes was investigated. The NK activity was estimated by means of radioactive chromium (51Cr) assay with the use of human leukemia cells K 562 as targets. The previous exposure of lymphocytes obtained from healthy donors to somatostatin in the concentration of 10(-8) and 10(-6) M resulted in the enhancement of NK activity. The finding provides a further evidence of the immunomodulating somatostatin action. PMID- 2567974 TI - A new class opioid peptide, [D-Arg2, beta-Ala4]-dermorphin tetrapeptide; physical dependence liability in mice. AB - Development of morphine-like physical dependence of the [D-Arg2, beta-Ala4] dermorphin tetrapeptide (H-Tyr-D-Arg-Phe-beta-Ala-OH) has been evaluated and compared with the physical dependence liability of morphine or pentazocine. Degree of the physical dependence was assessed by the naloxone-precipitated jumping behaviour in mice after treatment of a single dose of each compound. The number of jumps and the time of latency to first jump were recorded in this experiment. Number of jumps in a group pretreated with the peptide showed less than that in morphine-treated group. In addition, latency to the appearance of the first jump in the peptide-treated mice was later than that in the morphine treated group. The present results indicate that the physical dependence induced by [D-Arg2, beta-Ala4]-dermorphin tetrapeptide may be less marked than that produced by morphine. It is also interesting to note that the antinociceptive effect of this opioid peptide is more powerful and of longer duration than that induced by morphine or pentazocine. PMID- 2567975 TI - Anatomical distribution of somatostatin immunoreactivity in the infant brainstem. AB - The distribution of somatostatin-immunoreactive structures in the infant brainstem was investigated using the peroxidase-antiperoxidase technique. A wide distribution of somatostatin-immunoreactive cell bodies and fibers was observed throughout the brainstem. Numerous somatostatin-immunoreactive cell bodies and fibers were present in several areas of the brainstem including the substantia grisea centralis and the reticular formation. Some immunoreactive cell bodies were seen in cranial nerve nuclei such as the nucleus praepositus, the nucleus nervi hypoglossi and the vestibular nuclei. Immunoreactive fibers were seen in the nucleus cuneatus, the locus coeruleus, the nucleus tractus solitarius, the nucleus ambiguus, the nucleus tractus spinalis nervi trigemini and the dorsal horn of the spinal cord. These data were in agreement with previous works on the human adult. However, a high density of somatostatin-immunoreactive cell bodies and fibers in the interpeduncular nucleus and in the nucleus centralis superior, and a dense network of somatostatin-immunoreactive fibers in the dorsal part of the nucleus inferior olivarius, were also observed. The role of somatostatin in some brainstem nuclei and its probable implication in some specific neuropathological diseases of the infant brainstem is discussed. PMID- 2567976 TI - Prenatal ethanol exposure impairs lesion-induced plasticity in a dopaminergic synapse after maturity. AB - This study examined the consequences of alcohol (ethanol) exposure during fetal life on lesion-induced dopaminergic synapse responsiveness (plasticity) in the olfactory tubercle of the adult rat. Normally, in the olfactory tubercle, olfactory bulbectomy elicits alterations in pre- and postsynaptic dopaminergic markers, including, respectively, (1) increased tyrosine hydroxylase activity and immunoreactivity, which is associated with dopaminergic axon sprouting, and (2) increased dopaminergic receptor density and potentiated dopamine activation of adenylate cyclase. We have utilized biochemical and quantitative immunocytochemical methodology to examine these synaptic markers in olfactory bulbectomized or sham-operated adult rats. These animals were offspring of dams which were administered one of the following diets during pregnancy: (1) liquid diet containing 35% ethanol-derived calories ad libitum; (2) liquid diet containing an isocaloric amount of maltose-dextrin instead of ethanol, pair-fed; or (3) unaltered liquid diet ad libitum. The results show that prenatal alcohol exposure leads to suppression of the lesion-elicited dopaminergic synapse responsiveness in the olfactory tubercle. There were no significant differences between offspring born to control and pair-fed animals, indicating that the observed abnormalities were not due to alterations in their nutritional status. In conclusion, the present data are a biochemical and quantitative immunocytochemical demonstration of impaired lesion-induced synaptic responsiveness. This renders a new dimension in support of previous evidence indicating that prenatal alcohol exposure leads to altered neuroanatomical, neuroendocrinological and behavioral responsiveness to various challenges. Such impaired synaptic responsiveness may underlie brain functional abnormalities characteristic of fetal alcohol syndrome. PMID- 2567977 TI - Variability in antihypertensive drug therapy in general practice: results from a random national survey. AB - A random national survey of 50 general practitioners was undertaken to ascertain current trends in the pharmacological management of hypertension. Forty general practitioners entered the study, and scripts written by them over a two month period in 1988 were collected and recorded by the pharmaceutical pricing offices of the health department. The general practitioners provided details on whether the script had been written for hypertension, as well as the age and sex of the patients. Information from 37 general practitioners was available for study, involving 2675 scripts written for hypertension for 1858 patients. Sixty point two percent of the treated hypertensives were female, and 58.9% were aged over 60 years. The most commonly prescribed antihypertensives were the diuretics (47.1% of patients) and beta blockers (47.9%). They were followed by angiotensin converting enzyme inhibitors (18.2%) and calcium antagonists (9.7%). There was substantial variability in the prescription of antihypertensives with respect to the age and sex of the patients treated; the mean costs and duration of supply of different generic drug types; and the prescribing habits of general practitioners. By generic type, the mean monthly costs of therapy ranged from $3.77 (diuretics) to $48.19 (calcium antagonists). The age and sex adjusted geometric mean script costs ranged from $17.78 to $49.11 per month (median: $29.30). It seems unlikely that the observed degree of variability is explained by differences in the severity of hypertension between general practice populations. PMID- 2567978 TI - Host response to infection of a subperiosteal hydroxylapatite implant. AB - Particulate, nonresorbable hydroxylapatite is currently a popular implant material for the augmentation of atrophic alveolar ridges. Most reports have demonstrated favorable biocompatibility with common usage, but how the host bone will respond to the implant in the presence of an infection has not yet been investigated. Hydroxylapatite was implanted subperiosteally on one side of the mandible in four New Zealand White rabbits. After 3 months were allowed to elapse for stabilization of the implant, an infection was induced in both sides of each mandible by inoculation with Bacteroides melaninogenicus. One month later the animals were put to death and both sides of each mandible were examined microscopically. On the nonaugmented side there was total resolution of the infection. The side of each mandible containing the hydroxylapatite implant showed very mild chronic inflammation throughout the medullary space and periosteum. Although the number of animals used was small, the results suggest that the presence of the hydroxylapatite implant may have interfered with the host's ability to resolve the infection in the underlying bone. The possible mechanism of this interference is discussed. PMID- 2567980 TI - Designed diagnostic restriction fragment length polymorphisms for the detection of point mutations in ras oncogenes. AB - The polymerase chain reaction (PCR) technique has greatly facilitated the identification of ras oncogenes by allele-specific hybridization of the PCR amplified DNA to radioactively labelled oligonucleotide probes. In this study, we describe a different method which employs designed mismatch primers that create diagnostic restriction fragment length polymorphisms (RFLPs). This procedure allows the identification of point mutations in the amplified DNA without the use of any radioactive probes. We apply this method to the detection of specific point mutations in the rat H- and K-ras oncogenes in carcinogen-induced tumors. We also suggest strategies for the diagnostic RFLP analysis of most point mutations in the 12th and 61st codons of human ras oncogenes. This simple method is especially suitable for analyzing minuscule amounts of tissue samples where only a fraction of cells may carry activated oncogenes. PMID- 2567979 TI - Increase in ribosomal protein S6 phosphorylation is due to v-erbB-transforming activity and not to v-erbA mitogenic activity in avian erythroblastosis virus infected chicken embryo fibroblasts. AB - Avian erythroblastosis virus (AEV-ES4), a transforming avian retrovirus, transforms chicken embryo fibroblasts (CEFs) in culture and induces the maintenance of ribosomal protein S6 phosphorylation in the absence of serum. This effect is less pronounced after AEV-ES4 transformation than after transformation by Rous sarcoma virus (PR-RSV A). However, our results indicate that the two viruses induce an activation of the same S6 phosphokinase, as evidenced by the identity of S6 phosphopeptides and phosphoaminoacids in the two cases. Moreover this activation is performed through a protein kinase C-independent pathway. Expression of the v-erbA oncogene alone, which enhances the growth potential of CEFs, is not able to maintain S6 phosphorylation either in the absence of serum or in the presence of low serum concentration (0.5%). Expression of the v-erbB oncogene alone is responsible for all these AEV-ES4-induced effects. Furthermore, the maintenance of S6 phosphorylation in the absence of serum might be correlated with the degree of transformation of AEV-ES4-infected CEFs. These results show that S6 phosphorylation is one of the biochemical mechanisms deregulated by v erbB expression and is involved in the transformation process. PMID- 2567981 TI - [Biosynthetic growth hormone: current therapeutic status and perspectives]. AB - This paper covers the practical aspect of GH therapy in both GH deficiency and Turner syndrome. Dose regimen, route and frequency of administration and follow up are detailed. Prospectives of therapeutic use of growth hormone are discussed. PMID- 2567982 TI - Somatostatin: a hypothalamic transmitter for thermoregulation in rats. AB - The changes in both the thermoregulatory responses and brain somatostatin (SS) levels produced by ambient temperature (Ta) changes were assessed in rats after they had been equilibrated to each of the Ta for a period of about 90 min. Cold exposure, in addition to elevating hypothalamic SS-levels, led to increased metabolism and cutaneous vasoconstriction at Ta = 8 degrees C. In contrast, heat exposure, in addition to lowering hypothalamic SS-levels, resulted in decreased metabolism and cutaneous vasodilation at Ta = 30 degrees C. Rats were chronically implanted with a hypothalamic cannula to allow intrahypothalamic injection of SS on the conscious rats. Direct administration of SS (0.1-0.3 micrograms) into the preoptic anterior hypothalamic area caused a dose-related rise in colon temperature at three Ta tested. The SS-induced hyperthermia was produced by increased metabolism at Ta = 8 degrees C, whereas at Ta = 30 degrees C, it was caused by cutaneous vasoconstriction. At Ta = 22 degrees C, the hyperthermia was caused by increased metabolism and cutaneous vasoconstriction. Systemic administration of cysteamine, in addition to lowering hypothalamic SS-levels, produced a dose-related fall in colon temperature at Ta of 8 degrees C and 22 degrees C. The hypothermia induced by cysteamine was produced by decreased metabolism at Ta = 8 degrees C, whereas at Ta = 22 degrees C, it was caused by both decreased metabolism and cutaneous vasodilation. The data indicate that the hypothalamic SS-levels mediate normal body temperature responses in rats. PMID- 2567983 TI - An RFLP in mice detected by a CD3-epsilon probe. PMID- 2567984 TI - A polymorphic locus [D21S144] is detected by probe pVC12 on chromosome 21. PMID- 2567985 TI - Isolation and mapping of a polymorphic DNA sequence, DXS312, to Xq27----Xq28. PMID- 2567986 TI - A TaqI RFLP in Xq26----qter detected by pX301b [DXS311]. PMID- 2567987 TI - Nhe I and Hinc II polymorphisms in the human laminin B1 chain gene on 7q22. PMID- 2567988 TI - The gelsolin (GSN) cDNA clone, from 9q32-34, identifies BclI and StuI RFLPs. PMID- 2567989 TI - An XmnI RFLP detected with a cDNA probe for the CYP2C gene locus on chromosome 10. PMID- 2567990 TI - Discovery of a novel multilocus DNA polymorphism [DNF24]. PMID- 2567991 TI - [Eosinophil pleuritis induced by salazosulfapyridine]. PMID- 2567992 TI - Studies on the DNA elongation inhibitor and its proliferating cell nuclear antigen-dependent control in simian virus 40 DNA replication in vitro. AB - A 120-kDa protein that blocks DNA termini has been purified from extracts of HeLa cells. This protein inhibits the action of a number of enzymes that catalyze reactions involving the 5' and 3' ends of DNA (DNA ligase, 3' and 5' exonucleases, and DNA polymerase alpha). The 120-kDa protein blocks the synthesis of long DNA chains that are normally formed during simian virus 40 DNA replication, causing the accumulation of small DNA fragments. The effects of this protein can be reversed by the addition of proliferating cell nuclear antigen and other protein fractions (activators). PMID- 2567993 TI - Differential DNA sequence deletions from chromosomes 3, 11, 13, and 17 in squamous-cell carcinoma, large-cell carcinoma, and adenocarcinoma of the human lung. AB - Activation of protooncogenes and inactivation of putative tumor suppressor genes are genetic lesions considered to be important in lung carcinogenesis. Fifty-four cases of non-small-cell lung cancer (23 adenocarcinomas, 23 squamous-cell carcinomas, and 8 large-cell carcinomas) were examined for loss of DNA sequences at 13 polymorphic genetic loci. Loss of heterozygosity was seen more frequently in squamous-cell carcinoma than in adenocarcinoma. The loss of DNA sequences from the short arm of chromosome 17 (D17S1 locus) was detected in 8 of 9 heterozygous cases of squamous-cell carcinoma and in only 2 of 11 heterozygous cases of adenocarcinomas. Furthermore, in 7 of these 8 squamous-cell carcinomas, loss of heterozygosity from chromosome 17 was accompanied by loss of DNA sequences from chromosome 11. The spectrum of allelic sequences lost from chromosome 11 was, however, similar in every type of carcinoma studied, and the data show two regions commonly deleted from chromosome 11 (11pter-p15.5 and 11p13-q13) that may have a role in the pathogenesis of all these types of non-small-cell bronchogenic carcinoma. Loss of DNA sequences from chromosome 3 was seen in 16 of 31 cases where the constitutive DNA was heterozygous-i.e., informative. These data included only 6 of 16 cases where loss of heterozygosity involved a chromosomal locus previously shown to be lost consistently in small-cell lung cancer (DNF15S2). Loss of heterozygosity at the chromosome 13q locus, D13S3, was seen in 9 of 21 informative cases, and in 2 cases, both adenocarcinomas, duplication of the intact DNA sequences suggested the possibility that mitotic recombination had occurred. Frequent DNA sequence deletions, including those from chromosome 17, in squamous-cell carcinomas may reflect the extensive mutagenic and clastogenic effects of tobacco smoke that may lead to inactivation of putative tumor suppressor genes. PMID- 2567994 TI - Essential features of the P-glycoprotein pharmacophore as defined by a series of reserpine analogs that modulate multidrug resistance. AB - We have shown previously that reserpine is an effective "modulator" of P glycoprotein-associated multidrug resistance (MDR). In addition to enhancing drug cytotoxicity in our multidrug-resistant human leukemia cell line, CEM/VLB100, reserpine strongly competes with a photoactivatible analog of vinblastine, N-(p azido-3-[125I]iodosalicyl)-N'-(beta-aminoethyl)vindesine, for binding to P glycoprotein. We also demonstrated previously that there are three substructural domains present in many compounds that modulate P-glycoprotein-associated MDR: a basic nitrogen atom and two planar aromatic rings. In the present study, we wished to test more rigorously the hypothesis that not only are these domains necessary for modulators of MDR but also they must exist in an appropriate conformation. Reserpine is a modulator of MDR in which these domains are present in a well-defined conformation. Accordingly, we prepared eight compounds that vary the spatial orientation of these domains, using either naturally occurring reserpine or yohimbine as chemical templates. When tested for their ability to enhance the cytotoxic activity of natural product antitumor drugs in CEM/VLB100 cells, five compounds that retained the pendant benzoyl function in an appropriate spatial orientation all modulated MDR. By contrast, compounds lacking this moiety failed to do so. These active modulators competed strongly with the 125I-labeled vinblastine analog for binding to P-glycoprotein in plasma membrane vesicles prepared from these cells. Conformational analysis using molecular mechanics revealed the structural similarities of the active modulators. Our results support the hypothesis that the relative disposition of aromatic rings and basic nitrogen atom is important for modulators of P-glycoprotein-associated MDR, and they suggest a ligand-receptor relationship for these agents. These results also provide direction for the definition of an MDR "pharmacophore." PMID- 2567995 TI - Formation of nitric oxide from L-arginine in the central nervous system: a transduction mechanism for stimulation of the soluble guanylate cyclase. AB - A soluble enzyme obtained from rat forebrain catalyzes the NADPH-dependent formation of nitric oxide (NO) and citrulline from L-arginine. The NO formed stimulates the soluble guanylate cyclase and this stimulation is abolished by low concentrations of hemoglobin. The synthesis of NO and citrulline is dependent on the presence of physiological concentrations of free Ca2+ and is inhibited by NG monomethyl-L-arginine, but not by its enantiomer NG-monomethyl-D-arginine or by L canavanine. L-Homoarginine, L-arginyl-L-aspartate, or L-arginine methyl ester can replace L-arginine as substrates for the enzyme. These results indicate that NO is formed from L-arginine in the brain through an enzymic reaction similar to that in vascular endothelial cells, neutrophils, and macrophages, adding support to our hypothesis that the formation of NO from L-arginine is a widespread transduction mechanism for the stimulation of the soluble guanylate cyclase. PMID- 2567997 TI - Ultrastructure of Sertoli cells in scrotal testis of unilaterally cryptorchid goat. AB - Electron microscopy revealed changes in Sertoli cells of the scrotal testes of unilaterally cryptorchid West African dwarf goats from neonate to maturity. The major changes included alteration of the cell shape due to proliferation of the spermatogenic cells, development of nucleolar vesicles and indentations of nuclear outlines, and profusion of smooth endoplasmic reticulum. These alterations are similar to those described for the differentiation of Sertoli cells in normal testis. PMID- 2567996 TI - Visual responses in adult cat visual cortex depend on N-methyl-D-aspartate receptors. AB - We have investigated the role of the N-methyl-D-aspartate (NMDA) receptor, a subtype of glutamate receptor, in the responses of cells in adult cat visual cortex. After intracortical infusion of the NMDA receptor antagonist DL-2-amino-5 phosphonovalerate (DL-APV) for one day, iontophoretic responses to NMDA, to kainate, and to quisqualate revealed a receptor blockade specific to NMDA receptors and extending several millimeters from the cannula. In this region, neuronal responses to visual stimulation were profoundly suppressed, in a manner strongly correlated with the degree of NMDA receptor blockade. Neither NMDA receptor blockade nor activity suppression was caused by the inactive stereoisomer L-APV. Hence, we conclude that NMDA receptors make a major contribution to normal excitatory transmission in adult visual cortex. PMID- 2567998 TI - Seminal proteins binding to spermatozoa. PMID- 2567999 TI - Gene mapping and the muscular dystrophies. PMID- 2568000 TI - Effects of cysteamine and pantethine on open-field behavior, hypothalamic catecholamine concentrations, and somatostatin-induced barrel rotation in rats. AB - Cysteamine administered in a dose of 1.95 mM/kg subcutaneously (SC) markedly reduced several open-field behaviors (locomotion, rearing, grooming and defecation), while pantethine, administered in an equimolar dose, reduced the locomotion only. However, administered in a dose of 3.90 mM/kg (SC), pantethine also markedly reduced all open-field parameters. Cysteamine, and to less extent pantethine, reduced noradrenaline, and increased dopamine and DOPAC concentrations in the hypothalamus. It is discussed whether the lower potency of pantethine on open-field behaviors and hypothalamic catecholaminergic neurotransmission is connected with the limited activity of pantetheinase, the cysteamine-generating enzyme. Intracerebroventricularly (ICV) administered somatostatin did not influence the pantethine-induced (1.95 mM/kg SC) behavioral changes in the open-field test. It is possible that the peptide did not reach at the receptor sites in a sufficient concentration because of the reduced endogenous somatostatin content, or that the pantethine-induced noradrenaline depletion is connected with the ineffectiveness of somatostatin. Furthermore, pretreatment with cysteamine (1.95 mM/kg SC) or pantethine (1.95 mM/kg or 3.90 mM/kg SC) attenuated the somatostatin-induced (10 micrograms ICV) barrel rotation, suggesting that the level of endogenous somatostatin may play a role in the pathogenesis of this motor disturbance. PMID- 2568001 TI - MIF-1 is active in a chronic stress animal model of depression. AB - MIF-1 was tested in an animal model of depression that used unpredictable chronic stress. In this paradigm, rats received either no stressors or a daily protocol of a variety of stressors for 20 days, during which time daily, intraperitoneal injections of various compounds were given. The tricyclic antidepressant imipramine (5 mg/kg) and low doses (0.1 and 1.0 mg/kg) of MIF-1 significantly increased activity and decreased defecation in an open field on day 21. No dose of naloxone (0.01-10.0 mg/kg) acted as an antidepressant. A high dose (10.0 mg/kg) of MIF-1 significantly increased the effects of chronic stress and produced hyperalgesia. Chronically-stressed rats were significantly more analgesic than controls. The results indicate that MIF-1 can act as an antidepressant in this model. PMID- 2568002 TI - Anxiolytic effect of caffeine and caffeine-clonazepam interaction: evaluation by NaCl solution intake. AB - The administration of drugs with anxiolytic action to rehydrating rats augments the intake of 1.5% NaCl solution. In order to clarify the status of caffeine as an anxiolytic agent and its possible interaction with a benzodiazepine having high potency and efficacy in this regard, caffeine (0.78-100 mg/kg) alone and caffeine (0.78-50 mg/kg) plus clonazepam (0.05 or 0.50 mg/kg) injections (IP) were administered to rehydrating rats prior to 1-hr sessions during which they drank 1.5% NaCl solution. When given alone, caffeine, within a particular dose range, and clonazepam at both doses, augmented NaCl solution intake, but when administered in combination, caffeine antagonized the effects of clonazepam. PMID- 2568003 TI - The effects of beta-antagonists and anxiolytics on conflict behavior in the rat. AB - The present studies were designed to evaluate the effects of beta-adrenoceptor antagonists and traditional anxiolytics (phenobarbital and diazepam), alone and in combination, on behavior in the Conditioned Suppressioned of Drinking (CSD) conflict paradigm, an "animal model" for the study of anxiety and antianxiety agents. In daily 10-minute sessions, water-deprived rats were trained to drink from a tube which was occasionally electrified (0.5 mA), electrification being signalled by the presence of a tone. Within 2-3 weeks, control responding had stabilized (10-15 shocks/session and 10-15 ml water/session); drug tests were then conducted at weekly intervals. As expected, diazepam (0.6-10 mg/kg) and phenobarbital (10-40 mg/kg) administration resulted in a marked and dose dependent increase in punished responding at doses which did not markedly alter background responding (water intake). Neither propranolol (0.5-8 mg/kg) nor the beta-1-selective antagonist atenolol (1-16 mg/kg) significantly affected punished responding in the CSD. Both propranolol and atenolol produced significant beta-1 adrenoceptor blockade, as evidenced by the production of significant bradycardic effects in conscious rats at the doses employed. Pretreatment with 2.0 mg/kg propranolol did not alter the anticonflict effects of diazepam (0.6-10 mg/kg) or phenobarbital (10-40 mg/kg). Further, reduction of the shock intensity to 0.125 mA (i.e., decreased suppression) failed to alter the behavioral response to propranolol (1.5-5 mg/kg) or the interaction of 2.0 mg/kg propranolol with diazepam. Finally, chronic administration of propranolol (2.0 mg/kg, twice daily) did not affect punished responding over the course of 5 weeks of treatment. These data suggest that the CSD paradigm, although an effective "animal model" for the study of benzodiazepine and barbiturate anticonflict effects, cannot serve as an "animal model" for the study of the situation-specific (i.e., phobic) anxiety for which propranolol and related agents are presently used. PMID- 2568004 TI - BAEP screening for the prediction of tardive dyskinesia in schizophrenia. PMID- 2568006 TI - [Abdominal testicular retention with seminoma and epididymo-testicular separation]. PMID- 2568005 TI - Changes in eye tracking during clinical stabilization in schizophrenia. AB - Eye tracking abnormalities have been proposed as a trait marker for schizophrenia on the basis of their familial prevalence and the consistency of tracking over time in clinically stable patients. However, few studies have examined stability through acute episodes of illness, and most studies have not analyzed changes in different forms of eye movements. Therefore, the authors examined eye tracking, clinical state, and neuroleptic dose during 4 consecutive weeks in nine recently hospitalized schizophrenic patients. For the patients and controls, qualitative ratings of pursuit accuracy remained relatively stable over time. In contrast, saccade frequency increased significantly, with a 57% increase in small saccades and a 77% reduction in larger saccades. In comparison with cross-sectional studies which have found no correlation between neuroleptic dose and tracking performance, a reduction in large saccades was strongly correlated with increase in neuroleptic dose. The findings suggest that pursuit accuracy may be a trait characteristic of schizophrenia, while the frequency and size of saccades are state dependent characteristics. PMID- 2568007 TI - [Management of the hypersecretion of acid in Zollinger-Ellison syndrome (ZES)]. PMID- 2568008 TI - Localization of alpha 1-adrenoceptor in the rat kidney: in vitro autoradiography using [3H]-bunazosin. AB - We studied the precise localization of alpha 1-adrenoceptor in the rat kidney using in vitro macro- and micro-autoradiography (ARG) of [3H]-bunazosin. The slide-mounted nonfixed frozen sections (10 micron) of kidney were incubated with [3H]-bunazosin (5 nM) at 24 degrees C for 45 min, and then applied to macro- and micro-ARG. The macro-ARG revealed that the bunazosin binding sites were present mainly in the rat renal cortex. These bindings were evaluated to be specific by means of computerized image analysis system (RAS 1,000) of ARG. Micro autoradiogram showed that the tubules in the renal cortex had higher specific grain density than the glomeruli. These findings suggested that alpha 1 adrenoceptor in the rat kidney located mainly on the cortical tubules. PMID- 2568009 TI - Observations on gamma-glutamyl transferase, 5'-nucleotidase and leucine aminopeptidase activities in the plasma of the horse. AB - In 18 horses there was no effect of age or sex on plasma activities of gamma glutamyl transferase (gamma-GT), 5'-nucleotidase (5'-NT) and leucine aminopeptidase (LAP). All the enzymes were equally stable after storage for one month at -20 degrees C and there was no significant difference between their activities in serum and plasma in clinically normal horses. The pattern of release of gamma-GT, 5'-NT and LAP into plasma was studied in 114 horses which had a variety of orthopaedic, gastrointestinal, cardiovascular and hepatic (necrosis, lipidosis, neoplasia and cirrhosis) conditions. A definitive diagnosis of hepatic disease was established by histological examination of the liver. gamma-GT and 5'-NT were leaked into plasma in hepatic disease and gamma-GT was the more sensitive indicator of liver damage. There was some evidence that gamma GT and 5'-NT plasma activities may increase in hepatic necrosis as well as in biliary obstruction. LAP was insensitive and not hepatic specific in the horse. PMID- 2568010 TI - Urinary gamma-glutamyl transferase and the degree of renal dysfunction in 75 bitches with pyometra. AB - In 75 bitches with pyometra single urine samples were examined for gamma-glutamyl transferase (gamma-GT), protein, glucose, specific gravity, bacteria, red blood cells and white blood cells. Serum samples were examined for urea, creatinine, inorganic phosphate and gamma-GT. Biochemical findings were compared with the degree of illness (clinical signs). Twenty one bitches had no signs of renal disease. Seventeen showed only glomerular damage indicated by proteinuria without signs of proximal tubular damage. Thirty seven bitches had increased urinary gamma-GT levels, indicating proximal tubular lesions, which were associated with proteinuria in 35 and renal failure in 16 of them, and worse clinical findings. In all bitches with pyometra serum levels of gamma-GT were comparable to values in control bitches. Glomerular dysfunction seemed to precede proximal tubular lesions, so that gamma-GT-uria in bitches with pyometra was not an early but rather a late indication of a more profound degree of renal dysfunction, that is, proximal tubular renal damage developed after glomerular dysfunction and preceding renal failure. PMID- 2568011 TI - [Protection of ischemic myocardium]. PMID- 2568012 TI - [Shock caused by glafenine. Apropos of 7 complications]. AB - Glafenine-induced shock. Seven cases. Glafenine is a widely prescribed analgesic drug, and shock is one of its severe side-effects. We report six documented and one highly probable cases of such accidents. From these seven cases and a review of the literature, we have extracted the clinical characteristics of glafenine induced shock. Shock usually occurs about 30 minutes on average after taking one single tablet. Previous use of the drug is found in more than 50 per cent of the patients, and it was often followed by a neglected side-effect. A series of cutaneous and respiratory manifestations precedes or accompanies the shock. Two physiopathological mechanisms of glafenine-induced shock have been postulated: either anaphylaxis or idiosyncratic reaction involving the prostaglandins. In vitro tests give highly variable results and therefore are of limited value. Glafenine-induced shock is rare; its incidence, probably underestimated, is about 0.7 in 10(5) treatments. Prevention is essential, cure relies on adrenaline. PMID- 2568013 TI - [Therapy of acute and chronic urticaria and of Quincke's edema]. AB - Urticaria is a rather common, often vexing skin disease, characterized by evanescent, pruritic, erythematous wheals, and sometimes by giant hives (angioedema). The cause of chronic urticaria remains unknown in 75 to 80 percent of the cases. Its pathogenesis is related to the activation of tissue mast cells by many immunologic or non-immunologic mechanisms, resulting in a release of biologically active products. The therapeutic possibilities in practice are discussed. Non-sedative antihistaminic drugs of the H1 type (Terfenadine, Astemizole, Loratidine and Cetirizine) are the main stay in the treatment of urticaria. In case of failure the sedative non-selective antihistaminics from the old generation are used. Particular emphasis is given the possibilities to combining various antiallergic drugs. The combination of a H1 and H1 antihistamine can be effective in individual patients. PMID- 2568014 TI - [Prolonged hepatitis due to ajmaline--description of a case and review of the literature]. AB - A 60 years old woman was admitted for jaundice and fever which appeared after a treatment with ajmaline-butabarbital for two-weeks. Abdominal ultrasound examination and endoscopic retrograde cholangiography were normal. Mitochondrial antibodies were absent. Jaundice persisted for three years, associated with diffuse cutaneous xanthomatosis. Five years later, alkaline phosphatases remained elevated. A liver biopsy showed vanishing interlobular bile ducts with centrolobular cholestasis and ductular proliferation. We suggest that ajmaline can induce long lasting cholestasis due to damage to the intrahepatic bile ducts. The responsibility of butabarbital and the relationship with primary biliary cirrhosis are discussed. PMID- 2568015 TI - Treatment of reflux esophagitis with H2-blockers, antacids and prokinetic drugs. An analysis of randomized clinical trials. AB - Randomized clinical trials comparing H2-receptor antagonists, antacids, antacid/alginate and prokinetic drugs with placebo and other substances were analyzed. Symptomatic improvement has been shown for H2-receptor antagonists, antacids in very high doses and prokinetic drugs. H2-receptor antagonists, the present mainstay of reflux therapy, improve esophagitis, but they are insufficient in the treatment of severe esophagitis where omeprazole may become the therapy of choice. The evidence is weak that antacids, antacid/alginate, metoclopramide and domperidone heal esophagitis. PMID- 2568016 TI - Healing, relapse rates and prophylaxis of reflux esophagitis. AB - Gastroesophageal reflux is a daily occurrence in the general population. Reflux esophagitis is less common but still a considerable clinical problem. The results of medical therapy are generally clearly inferior to those seen in peptic ulcer disease. After healing relapse is rapid and maintenance has not been proved superior to placebo. The promising results with omeprazole (inducing pronounced acid inhibition) and surgery (strengthening anti-reflux mechanisms) indicate that a more aggressive approach may be needed in future treatment. Additional studies also using combinations of drugs both in the healing stage and during maintenance is needed. These should be compared to the long-term results of surgery. PMID- 2568017 TI - [Subdural empyema]. AB - Subdural empyema is a rare, but severe complication after skull-brain trauma and/or sinusitis. On the basis of four of our own cases the symptomatology, diagnostic problems and therapeutical management are discussed. Therapy should be an enlarged burr-hole trepanation or--if thick membranes develop--craniotomy and drainage of the empyema cavity. The use of modern CNS-penetrating antibiotics has canaed previously high mortality to decrease impressively. PMID- 2568019 TI - [Role of the adrenoreceptor apparatus in the mechanisms of bronchospasm and methods of diagnosing the types of adrenergic imbalance in patients with bronchial asthma]. PMID- 2568018 TI - Frequency and amplitude gradients of spontaneous release along the length of the frog neuromuscular junction. AB - Experiments were done at the neuromuscular junctions (NMJs) of the cutaneous pectoris muscle of the frog to test the validity of the "spatial decay method" which uses simultaneous recording with two intracellular electrodes, each located in the muscle fiber near a distal end of the terminal. The miniature endplate potential (MEPP) peak amplitude recorded by each electrode is used to calculate the position of the release site producing it as well as its initial amplitude in front of its release site. The validity of the method was tested with a third focal electrode located between the two intracellular electrodes, at about 60-100 microns from one of the intracellular electrodes, the probability of spontaneous transmitter release was not uniform along the NMJ either in low or in normal Ca2++ concentration. The release sites located in the region close to the first point of contact between the axon and the muscle fibre, usually close to the center of the NMJ, were spontaneously more active than the distal release sites. The degree of the steepness of proximodistal release gradients varied among different junctions. The less active regions tended to produce smaller MEPPs than did the more active ones. In fact, a correlation was found between the logarithm of MEPP frequency in a given region and the mean corrected MEPP amplitude in that region. These results also show that MEPP frequency was modulated to a greater extent than MEPP amplitude along the frog nerve terminal. The proximodistal gradient in MEPP frequency may be related to corresponding gradients in density and length of release sites along the junction, whereas MEPP amplitude gradients may be related to gradients in the length of release sites and/or postjunctional folds. PMID- 2568020 TI - Effects of quinoline and 8-hydroxyquinoline on mouse bone marrow erythrocytes as measured by the micronucleus assay. AB - Both quinoline and 8-hydroxyquinoline (HOQ) were tested for their genotoxicity in CD1 male mice by using a bone marrow micronucleus assay. Mice were intraperitoneally treated in single injections with three dose levels (25, 50, and 100 mg/kg) of each chemical with corn oil as solvent vehicle. Bone marrow was sampled at 24, 48, and 72 h postinjection. Quinoline resulted in a significant dose-related increase in the number of micronucleated polychromatic erythrocytes (MPCE) at the 24 h sampling time for all doses tested. The high dose (100 mg/kg) and the medium dose (50 mg/kg) also induced statistically significant increases (P less than .05) in the number of MPCEs at 48 h interval. The ratios of polychromatic to normochromatic erythrocytes at the 24 h sampling time were lower for the treated than the control animals. Although HOQ resulted in some increases in the number of MPCEs over the control, this compound induced a statistically significant increase in the number of micronucleated normochromatic erythrocytes (MNCEs) at all three doses following 24 h treatment. Both low and medium doses also induced a higher incidence of MNCEs at the 48 and 72 h sampling times. No data were available for the high dose at these times. The cytotoxic effect of this compound was expressed as low PCE/NCE ratios with all doses at 24 h after injection and as a high mortality rate in animals treated with the high dose (100 mg/kg). PMID- 2568021 TI - Developmental toxicity screen: results of rat studies with diethylhexyl phthalate and ethylene glycol monomethyl ether. AB - The purpose of these investigations was to develop a protocol for an in vivo developmental toxicity screen (DETS) that would provide sufficient data to determine whether to 1) do a full developmental toxicity evaluation without additional range-finding studies, or, depending on the results, to 2) do no further testing of a chemical. In order to evaluate this screen, we compared results obtained by using the DETS protocol with results of previously conducted developmental toxicity evaluations of diethylhexyl phthalate (DEHP) and ethylene glycol monomethylether (EGME). Five groups (n greater than or equal to 17) of F344 rats were treated on days 6-15 of gestation by dosed feed (DEHP levels = 0, 0.5, 1.0, 1.5, 2.0%) or gavage (EGME doses = 0, 12.5, 25, 50, 100 mg/kg/day). One half of the rats in these studies were killed on day 16 of gestation, and the remaining animals were allowed to deliver litters which were killed on day 4. EGME caused only a small decrease in maternal weight gain during treatment (100 mg/kg group) that was accompanied by a decrease in gravid uterine weight. The percentage of resorptions was increased in the 50 and 100 mg/kg groups. The number of live pups was decreased in the 25, 50, and 100 mg/kg groups, and litter weight and postnatal survival were decreased in the 100 mg/kg group. These results are consistent with those reported in developmental toxicity studies on EGME conducted by the inhalation and dermal routes. With DEHP, there were treatment-related reductions in maternal body weight and weight gain. There was also a nonstatistical increase in percentage of resorptions per litter that was also observed, but at relatively high levels, in a definitive study in which F344 dams were treated on days 0-20 of gestation. The results of studies on these two chemicals compare well with published results and would have led to the selection of proper dose levels for subsequent FDA segment 2 studies. PMID- 2568022 TI - Ozone and ultraviolet light act as additive cocarcinogens to induce in vitro neoplastic transformation. AB - Ozone, a major chemical oxidant in our environment, is an environmental air pollutant with putative carcinogenic action. Using in vitro transformation, we report for the first time that ozone (6 ppm for 10 min) acts in additive fashion with ultraviolet light (4 J/m2) to produce enhanced levels of transformation in hamster embryo cells and mouse C3H/10T-1/2 cells as compared to rats induced by each of the agents alone. The results underscore the hazard of ozone as a toxic pollutant which may have putative carcinogenic effects and interact with other environmental carcinogens. PMID- 2568023 TI - Toxic and DNA-damaging activities of the fungicides mancozeb and thiram (TMTD) on human lymphocytes in vitro. AB - The cytotoxic and mutagenic effects of the fungicides mancozeb and thiram were studied using human peripheral blood lymphocytes cultured in vitro with or without an S-9 mix microsomal metabolizing system. The results obtained suggested that the chemicals caused dose-dependent inhibition of thymidine uptake and unscheduled DNA synthesis on both resting and proliferating lymphocytes in the absence of the S-9 mix. In the presence of the S-9 mix, only thiram showed mutagenic activity by eliciting unscheduled DNA synthesis and a significantly higher frequency of sister chromatid exchanges than did controls. PMID- 2568024 TI - Limb bud cell culture for in vitro teratogen screening: validation of an improved assessment method using 51 compounds. AB - Rat embryo limb bud cells multiply and undergo chondrogenesis in micromass culture. Teratogenic agents are identified from their inhibition of chondrogenesis, which is quantified by determination of cartilaginous foci number or proteoglycan production. In other in vitro systems, the detection is based on their ability to affect cell proliferation. So far, these methods have failed to distinguish among true inhibition of differentiation, inhibition of cell proliferation, and nonspecific cytotoxicity. The improved technique involves simultaneous measurement of cartilage synthesis and cell multiplication. Differentiation was evaluated by measurement, using an Artek Counter, of nodule areas after Alcian blue staining and proliferation by spectrophotometric quantification of Crystal-Violet bound to micromass cells. Using this method, retinoic acid was shown to inhibit chondrogenesis without affecting cell multiplication, whereas 6-aminonicotinamide preferentially inhibited cell multiplication without affecting nodule size. Doxylamine (succinate), a known nonteratogen, induced inhibition of chondrogenesis, but with a parallel inhibition of cell multiplication, reflecting a nonspecific toxic effect. This improvement increases the specificity of the micromass culture test. Validation was performed using 51 compounds. Compounds were classified according to their inhibitory activity and their active concentration. The sensitivity of the test was 61%; the specificity, 100%; and the final accuracy, 75%. The method is fully miniaturised, automated, and computerised, allowing numerous compounds to be rapidly tested at very low cost. PMID- 2568025 TI - Genotoxic effects of estrogens in epithelial cells from the neonatal mouse uterine cervix: modifications by metabolic modifiers. AB - Epithelium from the neonatal mouse uterine cervix and uppermost part of vagina was cultured in vitro. The culture medium was supplemented with 17 beta-estradiol (E2; 10(-6)-10(-5) M) or diethylstilbestrol (DES; 10(-8)-10(-5) M) alone or in combination with different metabolic modifiers (alpha-naphthoflavone, beta naphthoflavone, phenobarbital, metyrapone, indomethacin) with postulated activating or inhibitory effects on DES metabolizing enzymes (cytochrome P-448- and P-450-dependent microsomal monooxidases, prostaglandin cyclooxygenase). E2 at 10(-5) M and DES at 10(-6) and 10(-5) M concentrations increased the incidence of cells with a high number of sister chromatid exchanges (high-frequency chromatid exchange cells, HFCEC). Indomethacin partially depressed DES-induced HFCEC, whereas the incidence was increased by alpha-naphthoflavone, which may be a result of stimulation of the fetal type of P-448-dependent enzyme activity or of DES increasing the metabolic activation of alpha-naphthoflavone. Phenobarbital and beta-naphthoflavone did not affect the incidence of DES-induced HFCEC. Metyrapone alone induced the highest incidence of HFCEC observed in this study, and this effect was inactivated by phenobarbital and/or DES. The mechanisms behind these results are discussed. This study shows that E2 and DES have a genotoxic effect (sister chromatid exchanges) in vitro in epithelial cells from the same target organ as in which epithelial aberrations occur after in vivo estrogen treatment in the neonatal period. The difference in incidence of tetraploid cells between stroma and epithelium is stressed (less than 5% vs. 16 47% depending on experimental group). PMID- 2568026 TI - DNA-DR typing shows HLA-DRw11 RFLPs are increased in frequency in both progressive systemic sclerosis and CREST variants of scleroderma. AB - DNA-DR typing of patients with both Progressive Systemic Sclerosis and CREST variants of Scleroderma has shown an increase in frequency of DRw11 RFLPs. The frequency of DQw7 was also increased in the patient groups, but this was due to linkage disequilibrium with DRw11. PMID- 2568028 TI - [Administration of intracorporeal, laser-induced shock waves for the destruction of urinary calculi]. AB - In spite of rapid developments in modern endourologic methods of urinary tract stone management, a definitive solution regarding the energy source for intracorporeal lithotripsy has not yet been found. Laser-induced shock wave lithotripsy is one of the most promising approaches. The evolution of this treatment modality within the last 4 years gives rise to the hope that a commonly accepted standard method will soon become available. General acceptance depends on increasing reliability and effectivity of current systems, especially in the case of problem stones. The advantages of intracorporeal shock wave lithotripsy can be summarized in the following points. (1) The shock wave is applied directly to the intended object, i.e., the stone, so that adverse side effects are virtually excluded. (2) Laser-induced shock wave lithotripsy leads to a precise and extremely fine fragmentation of stones, avoiding urinary tract obstruction with consecutive tract infection. (3) Cost and treatment handling are further points in favour of intracorporeal lithotripsy. (4) Perspectives are widened by the possibility of treating, of biliary, pancreatic and salivary gland duct stones. PMID- 2568027 TI - Effects of autonomic drugs on intraluminal pressure and excretion of rat seminal vesicles in vivo. AB - The effects of intravenously administered autonomic drugs on the intraluminal pressure and the excretion volume of the seminal vesicles in rats were studied. Both alpha 1-adrenergic and cholinergic stimulants produced a rapid increase in pressure and fluid excretion. alpha 1-Adrenergic stimulants elicited a sharp and single increase in pressure, while cholinergic stimulants caused a sharp increase with multiple peaks. PMID- 2568029 TI - Fatal cardiomyopathy: suspected child abuse by chronic ipecac administration. PMID- 2568031 TI - [Diagnosis and treatment of hypertrophic cardiomyopathy]. PMID- 2568030 TI - Sequences in E1A proteins of human adenovirus 5 required for cell transformation, repression of a transcriptional enhancer, and induction of proliferating cell nuclear antigen. AB - A range of deletion and other mutants in the coding region of the E1A gene of Ad5 has been assayed for transformation of baby rat kidney (BRK) cells in cooperation with ras, repression of the SV40 enhancer, and induction of proliferating cell nuclear antigen (PCNA). Transformation efficiency was drastically reduced by deletion of residues 4-25, 36-60, or 111-138 in exon 1 of the 289 residue (289R) and 243R E1A proteins. Deletion of other residues in exon 1 had little effect. With mutants in the region unique to the 289R protein, and in exon 2, the only effect on transformation seemed to be an increased tendency of mutant transformants, compared to wt, to migrate to form secondary foci. Repression assays, performed with E1A plasmids producing only the 243R protein, showed that deletion of residues 4-25 or 36-60 inhibited repression completely. Deletion of residues 128-138 reduced repression, but deletions elsewhere in exon 1 had little effect. Deletion of residues 188-204 in exon 2 reduced repression slightly, and deletion of all of exon 2 reduced it to about one-half. It is concluded that for transformation, there are two functional domains in E1A proteins, both in exon 1, both involved in binding different cellular proteins, and both probably concerned with different transforming functions. One of these domains, involving residues 4 25 and 36-60, also functions in repression, but the role of the second in repression is much less critical. All of the deletion mutants in exon 1 induced PCNA synthesis in BRK cells. This result, together with previously published work, suggests that the active site for PCNA induction either involves residues 61-69 or 82-85 in exon 1, which have not been deleted, or it does not depend on any single limited region of the E1A proteins. PMID- 2568032 TI - [The cyclic nucleotide system in various sections of the dog myocardium in experimental infarction]. AB - Distinct increase and then decrease in content of cyclic nucleotides was observed in dog myocardium ventricles and auricles within early periods of heart infarction (10 min-4 hrs). Within a day after ligation of artery the ratio cAMP/cGMP was considerably decreased as a result of activation of guanylate cyclase and cAMP-phosphodiesterase as well as due to a decrease in activity of adenylate cyclase. Acute ischemia of small area of the heart left ventricle caused impairment of cyclic nucleotide metabolism in all the heart muscle. PMID- 2568033 TI - [Effect of beta-adrenergic blockaders and nitrates on left ventricular diastolic function in patients with ischemic heart disease]. AB - By means of M-type electrocardiography the anapriline (propranolol--40 mg) and nitrosorbide (isosorbide dinitrate--10 mg) action on the left ventricular diastolic function was studied in the conditions of acute experiment in 55 patients with stable stenocardia. It was established that anapriline lengthens the diastole (at the expense of diastasis), decreases the myocardial relaxation speed and the fast filling and causes an increase of the telediastolic volume. Nitrosorbide decreases the filling velocity in the three diastolic phases without changing its duration and leads to a decrease of the telediastolic volume. In the combination of the two drugs anaprilin acts mainly on the duration, volume and speed indices of the left ventricular diastolic function while nitrosorbide acts mainly on the venous flow towards the heart and reduces the unfavourable action of anapriline on myocardial relaxation. PMID- 2568034 TI - Wound site as a predictor of complications following deep nail punctures to the foot. AB - The site of injury, condition of the nail, and type of foot covering were compared in 36 inpatients and 34 outpatients with nail puncture wounds to the foot. Of the 36 inpatients, 34 (94%) had pyarthrosis, osteomyelitis, or both. The plantar surface of the foot was divided into 3 zones. Of the 36 inpatients, 35 (97%) had deep puncture wounds in zone 1. In contrast, only 6 of 34 (18%) outpatients had injury to this area. Tennis shoes were shown to predispose to infection with Pseudomonas aeruginosa. Based on our findings, an early hospital admission should be considered for all patients with deep puncture wounds located in zone 1 and for patients who give a history of bone penetration in zone 2 or 3 at the time of injury. All patients who meet the above criteria and who are not admitted to hospital should be observed closely. PMID- 2568036 TI - [Personality characteristics and event related potentials (ERP)]. AB - Our investigation deals with high interindividual variability of Event-Related Potentials (ERP). We tried to find out systematic causes of these variability. In a performance situation 36 Ss were asked to solve arithmetical tasks. They got a weak electric shock applied at one finger when the result was wrong (negative feedback), at another finger when it was correct (positive feedback). The electrical stimulus triggered the ERPs. Special personality characteristics related to "need-achievement" (processing of success and failure) "and anxiety" had influence upon ERPs. We found various differences between ERPs following positive as well as negative feedback (success and failure) that point to the suggestion that there exist differences in processing feedback information in persons with different personality traits mentioned above. PMID- 2568035 TI - Possible famotidine-induced thrombocytopenia. PMID- 2568037 TI - [Proof of complex theories regarding conflicts, neuroses and their therapy, exemplified by Mjasiscew's theory of relations]. AB - Most psycho-dynamic theories emphasize the role of intra-personal conflicts in the aetiology and therapy of neurotic disorders, but very few can be empirically tested. The social psychological and psychodynamic "pathogenetic" theory of the "Leningrad School" of psychotherapy according to Mjasiscew (Lauterbach, 1984) maintains that conflicts between a person's relations with his social, material and cultural environment may be the cause of his neurosis. This theory is used to demonstrate that psychodynamic hypotheses about the role of intra-personal conflicts can be empirically tested with the help of "tests of conflict" (Lauterbach, 1987). PMID- 2568038 TI - [A dynamic model of loudness adaptation]. AB - Previous research on loudness adaptation has provided heterogeneous and partly contradictory results. One major reason seems to be the inconsistent use of the term 'adaptation', which is due to differing operational definitions. Here, a measurement-theoretic definition of loudness adaptation is introduced, which also comprises a new method for the identification of certain linear dynamic systems. This is of particular interest, since there is a lack of dynamic models describing loudness adaptation. An experiment with sinusoidal sound stimuli (3000 Hz) was performed to test whether loudness adaptation can be modeled by such a linear system. The results of the system identification indicate that this is possible. Therefore, it can be concluded that the given definition of loudness adaptation might be appropriate and that the method of system identification is an applicable research tool. PMID- 2568039 TI - [Brain abscess and subdural empyema from the bacteriologic viewpoint]. AB - The bacteriology has changed since beginning of antibiotic therapy. Reports of literature and the dates of patients treated in Neurosurgical Clinic of Munster are analyzed. The improvement of microbiological technique leads to the evidence of a greater role of anaerobics and of a smaller part of sterile cultures. The success of therapy depends on the early beginning of specific therapy. The poor prognosis of the sterile inflammations is caused by inefficiency of unspecific therapy. PMID- 2568041 TI - Reduction of secretion from a small-bowel fistula by somatostatin. Case report. AB - A 69-year-old man with intra-abdominal abscesses and small-bowel fistula after total gastrectomy received two 12-hour continuous i.v. infusions of somatostatin (250 micrograms/h) with a 24-hour interval. Secretion through the fistula decreased markedly during the infusion, with rebound rise after its discontinuation. The concentration of bilirubin in the secretion, but not that of amylase, also fell during somatostatin infusion. PMID- 2568040 TI - [Use of an immunoblotting method for identifying the individual proteins in the antigenic complexes of Bordetella pertussis]. AB - The composition of antigenic complexes isolated from the supernatant fluid of B. pertussis culture has been studied by means of immunoblotting techniques. In preparations obtained from B. pertussis strains 305 and 475 fragments of the molecule of fimbrial hemagglutinin, three subunits of B. pertussis toxin and agglutinogens 2 and 3 have been detected with the use of antisera to B. pertussis protective substances. PMID- 2568043 TI - Epidermis is the origin of high creatine kinase levels in skin blister fluid. AB - Creatine kinase (CK) isoenzyme, CK-BB, known as the brain fraction, is not normally present in serum but predominates in several normal and malignant tissues and body fluids. We recently reported increased CK-BB levels in suction blister fluid. In the present study the cellular origin of the enzyme in skin was studied from homogenates of blister top epidermis and blister base dermis as well as from homogenates of split skin dermatome shavings and isolated keratinocytes. The CK-BB in human skin was derived almost exclusively from the epidermis. Enzyme determinations from various spontaneous bullae suggest that all types of skin blisters initially contain high CK-BB levels. PMID- 2568042 TI - Biochemical and immunohistochemical comparison of collagen in granuloma annulare and skin sarcoidosis. AB - Collagen was studied by biochemical and immunohistochemical means in 5 patients with granuloma annulare (GA) and 3 with cutaneous sarcoidosis (SA). The solubility of collagen from the lesional skin in acetic acid was higher than that of collagen from unaffected skin from both patients and control subjects. Collagen concentration in the skin lesions, measured in terms of hydroxyproline content, was reduced in 3 patients with granuloma annulare and one with sarcoidosis, but the ratio of type III/I collagen was unchanged vis-a-vis non affected skin. The collagen concentration in non-affected skin of both GA and SA patients was also lower than in controls. The most typical immunohistochemical finding was the association of type III procollagen and fibronectin with granulomas in the lesional skin of both GA and SA cases. The activity of prolyl hydroxylase, a key enzyme in collagen biosynthesis, was markedly increased in the lesional skin, indicating that collagen synthesis in vivo was also increased. Surprisingly, collagen synthesis was not increased in cell culture studies. This could be due to cell selection as observed previously in scleroderma. Another possibility could be that various mediators released in vivo from inflammatory cells activate fibroblasts. However, when cells are subcultivated, this effect is not maintained. In conclusion, marked changes in collagen could be observed in granuloma annulare and skin sarcoidosis, reflecting increased turnover of collagen in vivo. PMID- 2568044 TI - Adenylate cyclase activity in homogenates of human melanoma cells. Effect of alpha-MSH and isoprenaline. AB - The effects of the alpha-melanocyte-stimulating hormone (alpha-MSH) (10(-7)-10( 5) M) and the beta-adrenoceptor agonist isoprenaline (10(-9)-10(-4) M) on adenylate cyclase (AC) activity were investigated in homogenates of the human IGR 1 melanoma cells with or without additional GTP. Basal AC activity was increased by the administration of 10 microM GTP. Alpha-MSH had no effect on cyclic AMP (cAMP) accumulation, while isoprenaline stimulated AC activity in a dose dependent manner. PMID- 2568045 TI - Expression of lymphocyte activation markers in benign cutaneous T cell infiltrates. Discoid lupus erythematosus versus lichen ruber planus. AB - The expression of lymphocyte activation markers (IL2 receptors, transferrin receptors and HLA-DR) was examined in cutaneous lymphoid infiltrates of 12 patients with lichen ruber planus (LP) and 10 individuals with discoid lupus erythematosus (DLE). The cell infiltrates in both conditions were generally of considerable size. The vast majority of the infiltrating cells were T cells. The reactivity of the anti-IL2 receptor antibody used was confined to lymphocytes. In patients with LP 26 +/- 17% of the infiltrating cells were IL2 receptor positive, 20 +/- 8% carried transferrin receptors and greater than 90% HLA-DR. In patients with DLE less than 1% were IL2 receptor positive, less than 5% carried transferrin receptors and greater than 90% were HLA-DR positive. These data indicate that IL2 receptor expression distinguishes the infiltrating T lymphocytes in LP and DLE, although in both conditions the vast majority of the infiltrating cells were activated as revealed by their expression of HLA-DR. PMID- 2568046 TI - Systemic effect of ultraviolet irradiation on non-immunologic immediate contact reactions to benzoic acid and methyl nicotinate. AB - Systemic effects of ultraviolet irradiation B (UVB) and ultraviolet irradiation A (UVA) on non-immunologic immediate contact reactions (NIICRs) induced by benzoic acid (BA) and methyl nicotinate (MN) were studied in healthy volunteers. NIICR tests with four concentrations of BA and MN in white petrolatum were performed on the skin of the upper back on exposed and non-exposed areas, before and at intervals 1-14 days after exposure to 1) 0.20 J/cm2 of UVB, 2) after the first of daily doses on five consecutive days of 0.04 J/cm2 of UVB, 3) after the first of daily doses on five consecutive days of 20 J/cm2 of UVA, and 4) 1-8 weeks after the first of twelve consecutive doses of 20 J/cm2 of UVA given three times a week over a period of four weeks. After the last of twelve exposures of UVA, stratum corneum was stripped off from both the exposed and the non-exposed skin. Forty minutes after application of the test substances, erythema and edema reactions were observed visually, and changes in the blood flow were monitored using a laser-Doppler flow-meter. All dosages of UV-light inhibited the NIICRs on exposed areas. UVB as given repeatedly inhibited NIICRs to 125 mM BA on non-exposed areas. The twelve doses of UVA also had a systemic inhibitory effect on NIICRs both on stripped and non-stripped test areas. The results indicate a systemic inhibitory effect of UV light on NIICRs. PMID- 2568047 TI - Cross-sensitization patterns in guinea pigs between cinnamaldehyde, cinnamyl alcohol and cinnamic acid. AB - Guinea pig maximization tests (GPMT) were performed with cinnamon substances. There was a certain degree of cross-reactivity between cinnamaldehyde, cinnamyl alcohol and cinnamic acid as animals sensitized to cinnamaldehyde reacted to the challenge with the three substances. Animals sensitized to cinnamyl alcohol reacted to cinnamyl alcohol and cinnamaldehyde, but not to cinnamic acid. Cinnamic acid did not sensitize guinea pigs. Compared to the challenge concentration for cinnamaldehyde, approximately a 15 times higher concentration of cinnamyl alcohol and a 25 times higher concentration of cinnamic acid were required to give positive reactions in animals sensitized to cinnamaldehyde. This could not be explained by differences in permeability properties, as the penetration profiles of the three substances through guinea pig skin in vitro showed permeability coefficients of the same order of magnitude under the test conditions. The study suggests that cinnamaldehyde is the "true" allergen, while cinnamyl alcohol and cinnamic acid are transformed in the skin to cinnamaldehyde, before contact allergic reactions can occur. PMID- 2568048 TI - Two cases of autoimmune progesterone dermatitis. Immunohistochemical and serological studies. AB - Two cases of autoimmune progesterone dermatitis are reported. The patients developed recurrent pruritic erythematous and edematous eruptions on the extremities, trunk or face, with occasional vesicles on the palms and soles. The eruptions appeared 7 to 10 days prior to their menstruation and persisted for several days. They showed immediately positive skin tests with 0.1 mg/ml and 0.2 mg/ml of aqueous progesterone suspension, respectively. The patients had IgG serum factor which bound rat corpus luteum. Positive indirect basophil degranulation tests against progesterone were demonstrated in both patients. Circulating autoantibodies to patients' own progesterone may cause or modulate the intermittent eruptions of the disease. PMID- 2568050 TI - 'Dry' skin in atopic dermatitis. II. A surface profilometry study. AB - In patients with atopic dermatitis, the skin is often 'dry' on non-eczematous areas, and feels rough to the touch. In the scanning electron microscope (SEM) this roughness corresponds to a change in the skin surface, from regular major and minor furrows into a coarse and irregular pattern. In the present study the topography of normal skin and 'dry' atopic skin was quantitatively recorded using a profilometry method. A computer-based three-dimensional reconstruction of the skin surface was made from replicas, and common roughness parameters were calculated. It was found that in the 'dry' atopic skin roughness parameters Ra, Rq, and Rmax, were significantly increased, whereas the number of peaks, Rn, was decreased. Parameters describing the shape (skewness, kurtosis, and delta a) of the profile were not significantly different. These topographical data clearly support the visual impression of the surface pattern of 'dry' atopic skin found in SEM. PMID- 2568049 TI - 'Dry' skin in atopic dermatitis. I. A clinical study. AB - A common finding in patients with atopic dermatitis is the occurrence of 'dry' skin on non-eczematous regions. 'Dry' skin is here defined as a clinical condition meaning a rough, finely scaling non-inflamed skin surface. The frequency and extension of 'dry' skin were examined in 50 patients with atopic dermatitis and were compared with those in 50 non-atopics. A discrepancy was found in both groups between the subjective opinion of the presence of 'dry' skin and the objectively noted 'dry' skin. Among the atopics, 48% were found to have 'dry' skin compared with 14% among the controls (p less than 0.01). The most frequent location of 'dry' skin in both groups was the back. Intolerance to wool was found to be significantly high (p less than 0.01) in the atopic group, although it was also quite common in non-atopics. In order to correlate the clinical observation to skin morphology, a replica-technique was used to visualize the surface of 'dry' skin in the scanning electron microscope. PMID- 2568051 TI - A double-blind study of ultraviolet phototherapy in the prophylaxis of chilblains. AB - A randomized, double-blind study was carried out to assess the prophylactic value of ultraviolet irradiation in the autumn as a means of preventing the development of chilblains on the toes and fingers during the course of the winter. Placebo irradiation was achieved by means of an optical filter which absorbed all ultraviolet radiation from the lamps but allowed the visible light component to be transmitted, thus giving patients the impression that both limbs were being treated. Patients were reviewed at monthly intervals during the winter. The response between patients was variable; some patients developed chilblains whilst others remained symptom free. However, in no patient did the ultraviolet treated limb differ from the untreated limb. We conclude that the ultraviolet phototherapy is of no value in the prophylaxis of chilblains. PMID- 2568052 TI - Etiology of vitiligo. A new hypothesis. AB - Serum from actively repigmenting human vitiligo subjects had maximum mitogenic effect on the growth of melanocytes in culture, followed by the serum from normal donors, and from untreated vitiligo subjects in that order. Based on these findings, a new hypothesis is suggested for the etiology of vitiligo. PMID- 2568054 TI - Acitretin induces an increased adherence of S. aureus to epithelial cells. AB - Recently, synthetic retinoids have been implicated as causing a rise in the incidence of staphylococcal infections in patients orally treated with these compounds for various disorders of keratinization. Since the adherence of bacteria to epithelia is an important early event in the development of bacterial infections, in the present study we investigated the in vitro effects of acitretin on the adherence of Staphylococcus aureus to epithelial cells of the anterior nares of 15 healthy human subjects. It was found that pre-incubation of nasal epithelial cells with acitretin causes a statistically significant (p less than 0.001) increase in the adherence of S. aureus to these cells, as compared to the controls. The growth of S. aureus cultures in the presence of acitretin exerted no effect on the staphylococcal adherence. These results suggest that the oral acitretin-induced increase in S. aureus colonization and in the incidence of cutaneous staphylococcal infections may be related to the enhancement of staphylococcal adherence to epithelia caused by this compound. PMID- 2568053 TI - Effects of single application of a moisturizer: evaporation of emulsion water, skin surface temperature, electrical conductance, electrical capacitance, and skin surface (emulsion) lipids. AB - Effects of single application of an oil in water emulsion were studied on the forearm skin of 12 healthy volunteers. Five different non-invasive methods were used. Values were followed for 360 min after application of the emulsion, with the contralateral forearm as untreated control. The evaporation of emulsion water from the skin surface immediately rose to high values, but within 15 min returned to the original level. A parallel initial increase in conductance was observed; however, this was followed by a slightly increased level throughout the 360 min study. Electrical capacitance was also slightly increased throughout the study. Skin surface lipids, dominated by emulsion lipids, were increased, with high values for at least 120 min, followed by a gradual decline toward normal. Single application of emulsion is characterized by an initial evaporation phase, with evaporation of emulsion water, which lasts less than 15 min, followed by a lipidization phase, which lasts at least 360 min, dominated by the oil constituent of the emulsion undergoing epidermal absorption. During the lipidization phase, epidermal hydration parameters are slightly but consistently improved. PMID- 2568055 TI - Seborrhoeic dermatitis and Pityrosporum ovale: a cultural and immunological study. AB - Seborrhoeic dermatitis is associated with Pityrosporum ovale, but the exact role of the organism is not clarified. In order to study this connection we have investigated 30 patients with seborrhoeic dermatitis with quantitative culture for P. ovale, serum IgG antibodies against P. ovale and lipid measurements. We compared the patients with 60 healthy individuals and found no significant difference in the number of P. ovale or serum antibodies. The lipid content on the skin was significantly higher in the patient group (p = 0.0001). There was no difference in the number of P. ovale in lesions compared to healthy skin in the patient group. This study support our theory that an abnormal reaction in the skin to P. ovale causes the inflammation and the number of P. ovale is of minor importance. PMID- 2568056 TI - An evaluation of broad-spectrum sunscreens against topical PUVA-induced erythema. AB - Protection against topical PUVA with broad-spectrum sunscreens was investigated. A protection factor against topical PUVA was established for broad-spectrum sunscreens against topical PUVA-induced erythema. PMID- 2568057 TI - Longitudinal melanonychia after healing of lichen planus. AB - A patient with progressive longitudinal ridging, diffuse pigmentation and narrowing of the finger nails as the only signs of lichen planus was treated with intramuscular injections of triamcinolone for 2 years. Her nails healed, with the appearance of normal proximal nails after 6 months. After one year, longitudinal bands of melanonychia appeared on the thumbs and still persist. PMID- 2568058 TI - Nail-changes induced by penicillamine. AB - Peculiar nail-changes in a 70-year-old woman with rheumatoid arthritis occurring after approximately 1 year of penicillamine treatment are described. After cessation of treatment there was a gradual resolution with regain of normal nails after 7 months. Reinstitution of penicillamine treatment caused a recurrence thus proving a causal relationship between penicillamine and the described nail changes. The fingernails were more affected than the toenails and clinically the changes consisted of absence of lunulae, longitudinal ridging, transverse or longitudinal defects of the nailplate and a tendency of onychoschizia. PMID- 2568059 TI - Localized bullous pemphigoid, a T cell-mediated disease? Electron microscopic and immunologic studies. AB - Two new cases of the rare, nonmucosal and nonscarring localized variety of pemphigoid are described. With reference to the data in the recent literature, the disease was classified as Localized Bullous Pemphigoid (LBP). The ultrastructural and immunologic findings are described and are briefly discussed in the context of the possible mechanisms leading to local subepidermal tissue injury in LBP. Although classical features of humoral responsiveness (in vivo bound IgG and complement at the epidermal basal membrane-zone (BM-zone)) were observed, a possible additional role of T cell mediated immunity in generating this local disease is considered. PMID- 2568060 TI - Acitretin monotherapy in acrodermatitis continua Hallopeau. AB - In a patient affected with acrodermatitis continua Hallopeau, acitretin (Ro 10 1670) monotherapy resulted in a complete clearance of pustulation at a dosage of 45 mg per day. At this dosage the leukotriene B4-induced intraepidermal accumulation of polymorphonuclear leukocytes was markedly inhibited. PMID- 2568061 TI - Treatment of hyperhidrosis manuum by tap water iontophoresis. AB - In a randomised, double-blind, controlled clinical trial of the effect of treatment with tap water iontophoresis, 11 patients with palmar hyperhidrosis were treated actively on one hand and with placebo on the other. The patients' sweat production was 100% higher (median) than measured in control subjects of the same age and sex. Prior to iontophoresis, the patient's sweat production was the same on both hands but after treatment it was reduced significantly on the treated hand (p less than 0.01) compared with the sweat production prior to treatment as well as with that of the untreated side. An 81% reduction (median) in sweating was found in 6 patients receiving maintenance treatment every second week. PMID- 2568062 TI - Short-contact therapy for psoriasis with 3.9% butantrone (10-butyryl dithranol). AB - Previous studies have shown that when butantrone and dithranol were used in equimolar gradually increasing concentrations in short-contact therapy for psoriasis the efficacy of butantrone was somewhat lower compared to dithranol. To see whether the efficacy of butantrone in short-contact therapy could be increased by starting with a single high-concentration directly, 20 psoriatic patients were treated with dithranol (0.1,-0.5,-1.0,-2.0%) and butantrone (3.9%) short-contact therapy as a right-left comparison. With these treatment modalities the antipsoriatic effects of dithranol and butantrone were similar. Although the efficacy of 3.9% butantrone was better than the previously used butantrone therapy with gradually increasing doses, there was a parallel increase in side effects. In general, the side-effects (erythema and staining) remained weaker on the butantrone-treated side than on the dithranol-treated side. No systemic adverse-effects were observed in any of the treated patients. PMID- 2568063 TI - Treatment of psoriasis and other dermatoses with a single application of a corticosteroid left under a hydrocolloid occlusive dressing for one week. AB - Lotions of five different corticosteroids were applied on 3 X 4 cm areas of large infiltrated chronic psoriatic plaque in eight patients, and left occluded with a hydrocolloid dressing (Actiderm) for one week. Area treated with clobetasol, betamethasone and triamcinolone were clinically healed within one week in four patients, but slight residual erythema was observed in the other four. Hydrocortisone-treated areas showed better improvement than the non-steroid treated control area, where infiltration and erythema remained unchanged. In 44 patients with psoriasis, lichen planus, chronic lichenified eczema, discoid lupus erythematosus and necrobiosis lipoidica, treatment with betamethasone applied 1-3 times under Actiderm once a week healed the lesions. The skin-coloured Actiderm is easy to apply and wear and does not loosen when taking a shower or hot bath. PMID- 2568064 TI - Treatment of psoriasis with a new UVB-lamp. AB - Twenty-nine patients with psoriasis took part in a study which compared the therapeutic effect of Philips TL12 with a new narrow-band UVB lamp (Philips TL01). The patients were treated on an out-patient basis and treatment were given 3-5 times weekly for a maximum of 8 weeks. The study was conducted in a randomized left-right double-blind fashion. The total score on the TL01-side decreased rather more than on the TL12-side. Eleven patients preferred the TL01 lamp and one the TL12-side, whereas 17 patients had no preference. One of the drawbacks with the new lamp is that radiation times are almost doubled. PMID- 2568065 TI - The effect on atopic dermatitis of supplementation with selenium and vitamin E. AB - Reduced concentrations of selenium in whole blood, plasma and white cells and reduced activity of selenium-dependent glutathione peroxidase in red cells have been found in atopic dermatitis. To determine the effect of selenium supplementation on this disease, the normal daily diet of 60 adults with atopic dermatitis was supplemented with selenium-enriched yeast for 12 weeks in a randomised double-blind study. Group 1 took 600 micrograms of selenium alone, Group 2 600 micrograms of selenium plus 600 IU of vitamin E and Group 3 a placebo. After 12 weeks, there was a significant increase in the concentration of selenium in whole blood and the activity of selenium dependent glutathione peroxidase in platelets in Groups 1 and 2 and the concentration of vitamin E in plasma in Group 2. There was no significant difference between the three Groups in the severity of the eczema or the concentration of selenium either before or after the 12 weeks of supplementation. The results suggest that although selenium enriched yeast supplement was absorbed and bioavailable it does not enter the skin or produces a worthwhile improvement in atopic dermatitis. PMID- 2568066 TI - Local anaesthesia with a lidocaine/prilocaine cream (EMLA) for cautery of condylomata acuminata on the vulval mucosa. The effect of timing of application of the cream. AB - The analgesic efficacy of a lidocaine/prilocaine cream (EMLA) for the cautery of genital warts was evaluated in an open study. Fifty-two women aged 18 to 28 with at least two condylomata on the vulval mucosa took part. In a pilot study (n = 10) the time of onset of anaesthesia after the application of EMLA to the mucosa was established by pinching with a forceps. All ten patients were anaesthetized within 5-7 min and cautery was performed with no or only slight pain in 9/10 patients. In the main study EMLA was applied to the mucosa of 42 women for 10, 15 or 20 min. The anaesthesia was satisfactory for the cautery of condylomata in 92% of the patients after the application of EMLA for 10 minutes. The analgesic efficacy decreased gradually with application times of 15 min or longer (p less than 0.05). In the case of insufficient anaesthesia, an additional application of EMLA for 2-5 min enabled the operations to be completed in 7/8 patients. PMID- 2568067 TI - Cutaneous cryptococcosis resembling molluscum contagiosum: a first manifestation of AIDS. AB - A 30-year-old homosexual man developed multiple skin umbilicated lesions resembling molluscum contagiosum. Initially the lesions were on his face but they rapidly spread. Histopathology and mycologic cultures of a skin biopsy revealed cryptococcus neoformans which was also identified in cerebrospinal fluid and in bronchoalveolar washings. The patient had fever, weight loss, generalized lymph node enlargement, depletion of the T helper subpopulation and positive HIV-1 serology. During treatment with flucytosine and amphotericin B, the skin lesions regressed in 3 months (cryptococcus neoformans disappeared in the cerebrospinal fluid and skin within one and five weeks, respectively). Our case demonstrates that molluscum contagiosum-like skin manifestations may be caused by cryptococcal infections. So it is necessary to perform skin biopsy in HIV seropositive patients with skin lesions resembling molluscum contagiosum, to diagnose mycotic infections, and especially cryptococcosis. Cutaneous cryptococcosis was, in this case, the first symptom of AIDS. PMID- 2568068 TI - Immunohistochemical investigations of beta-endorphin in human pancreatic islets. AB - The PAP technique was used to examine adult human pancreata (corpus) immunohistochemically for the presence of beta-endorphin containing cells. These cells were found to account for 4.8% of the islet cells. They are irregularly distributed within the islets, where they occur singly or in groups of 3 to 5 cells between the acini (0.4% of the parenchyma). Investigations designed to detect the simultaneous presence of beta-endorphin and somatostatin or glucagon revealed that beta-endorphin occurs in somatostatin cells (1.0% of the islet parenchyma). This is the 1st proof that these 2 hormones appear together. The simultaneous presence of beta-endorphin and glucagon in the same cell was also observed in 0.9% of the islet parenchyma. Earlier studies undertaken by us have shown that beta-endorphin is synthetized in the islets of Langerhans. Possible functions of beta-endorphin in the islets are discussed. PMID- 2568069 TI - Neurotransmitter deficits in a non-multi-infarct category of vascular dementia. AB - Brain tissue from 9 severely demented patients cared for in psychiatric long-term wards and with records of stroke episodes, macroscopic signs of brain infarcts and with no clinical evidence of senile dementia of the Alzheimer type was investigated and compared with control material. The mean volume of the brain infarcts in this vascular dementia group was only 6.8 ml. Pronounced disturbances of the serotoninergic and cholingergic systems were found in subcortical and cortical grey matter. These widespread neurotransmitter changes can hardly be explained by the localized brain infarcts per se, but suggest the existence of another category of vascular dementia. Since the neurotransmitter disturbances were found to be similar to those of Alzheimer's disease and senile dementia of the Alzheimer type, it seems more likely that they indicate a common pathway for dementia disorders than that they serve as markers of different dementia categories. PMID- 2568070 TI - Relationship between acupuncture-induced immunity and the regulation of central neurotransmitter system in rabbits--II. Effect of the endogenous opioid peptides on the regulation of acupuncture-induced immune reaction. AB - By injecting Naloxone, the T-lymphocyte transformation was reduced markedly. When the Naloxone plus electro-acupuncture were used together, it was not reduced but was still lower than it was when only electro-acupuncture was used. This explained why Naloxone could reverse the function by which electro-acupuncture promoted the T-lymphocyte transformation, and the endogenous opioid in the brain promoted the immune reaction. Through promoting the release of the endogenous opioid in the brain, electro-acupuncture raised the T-lymphocyte transformation function. Using electro-acupuncture when naloxone antagonized morphine, the T lymphocyte transformation function was still promoted markedly. The mechanism could be that electro-acupuncture excited the function of the endogenous opioid system in the brain. In acupuncture immune reaction, the relationship between the endogenous opioid and catecholaminergic systems was explored. PMID- 2568071 TI - Copper vapor laser and optical fiber catheter system for liquefaction and removal of thrombus in occluded arteries. AB - An instrument consisting of a copper vapor laser coupled to an optical fiber/chemical injector catheter for the treatment of occluded arteries has been constructed and tested. The combined application of three steps: the pre irradiation injection of a light absorbing dye, HPD; brief copper laser irradiation (at 578 nm); and a urokinase infusion after the irradiation, produced the striking effect of liquefaction and resolution of thrombus. The histological examination of the arteries after the treatment showed no apparent damage to the arterial wall. PMID- 2568072 TI - The effects of cranial TENS on measures of autonomic somatic and cognitive activity. AB - Peripheral and central nervous system effects of the cranial application of sub threshold transcutaneous electrical nerve stimulation (TENS), of sinusoid waveform passed between earlobe electrodes at an AC frequency of 100 Hertz, were investigated. In a single-blind study, each of thirty healthy volunteer subjects was administered one 30 minute treatment of either active TENS, placebo TENS or no treatment. Pretreatment to post-treatment changes in measures of autonomic activity (blood pressure, pulse rate, peripheral vasomotor activity), somatic activity (skeletal muscle tension), and anxiety were evaluated. Significant reductions in systolic blood pressure (p less than .05), diastolic blood pressure (p less than .01), pulse rate (p less than .05), peripheral vasomotor activity (marginally significant: p less than .07) and anxiety (p less than .05) were observed subsequent to active. TENS as compared to both placebo TENS and no TENS. No significant placebo TENS effect was observed. Possible mechanisms of action of this form of cranial TENS on the peripheral and central nervous system are discussed. PMID- 2568073 TI - Accelerated healing of skin ulcers by electrical stimulation and the intracellular physiological mechanisms involved. AB - Evidence is reviewed (8 studies involving 215 clinical patients with ischemic skin ulcers and 7 animal tissue or tissue culture studies) that electrical stimulation of fibroblast cells accelerates the intracellular biosynthesis necessary to form new granulation tissue in a healing wound, and that both a direct local tissue effect and a circulatory improvement occur. A model is presented in which transmembrane currents open voltage-controlled calcium channels in fibroblast cells, causing ATP resynthesis, activation of protein kinase mechanisms to synthesize new cellular protein, and the DNA replication necessary for mitotic cell division. Stimulation efficacy appears to be determined by a number of basic electrical parameters, and judicious waveform control is desirable. PMID- 2568074 TI - Unique changes found on the Qi Gong (Chi Gong) Master's and patient's body during Qi Gong treatment; their relationships to certain meridians & acupuncture points and the re-creation of therapeutic Qi Gong states by children & adults. AB - Changes taking place in both Qi Gong Masters and their patients during Qi Gong treatment were evaluated using the Bi-Digital O-Ring Test. During the Qi Gong state, on the Qi-Gong Master's body, as well as the body of the patient being treated, acupuncture points CV5 (Shi Men) and CV6 (Qi Hai)-- located below the umbilicus-- show changes from +4 in the pre-Qi Gong state to between -3 and -4 during the Qi Gong state. Before and after the Qi Gong, there is a normal +4 response to the Bi-Digital O-Ring Test at these acupuncture points. Similar changes were also observed on acupuncture points CV17 (Shan Zhong), CV 22 (Tian Tu), Yin Tang (at an area just between the eyebrows: the pituitary gland representation area, colloquially known as the "third eye") and GV20(Bai Hui), the entire pericardium meridian & triple burner meridian, their acupuncture points, the adrenal glands, testes, ovaries and perineum, as well as along the entire spinal vertebrae, particularly on and above the 12th thoracic vertebra, medulla oblongata, pons, and the intestinal representation areas of the brain located just above and behind the upper ear. Using these findings as criteria for evaluating the effectiveness of reaching the Qi Gong state, we were able to reproduce during the experimental trials similar changes in ourselves and the patient being treated with therapeutic effects comparable to those of the Qi Gong Master. Beneficial effects of external Qi Gong treatment given by a Qi Gong practitioner 1 to 3 times for 10-20 seconds each (although most Qi Gong masters take 3-20 minutes per treatment) often resulted in improvement of circulation and lowering of high blood pressure, as well as relaxation of spastic muscles, relief of pain, and enhanced general well-being, all of which resemble acupuncture effects. In order to reproduce the same procedure with others, we selected 4 children ranging between 8 and 11 years of age who had no knowledge of Qi Gong or Oriental medicine. One of these four children, the 8 year old, was able to consistently reach the same Qi Gong state after less than a half day and another child, 11, after less than 2 days. Within a week, the other two were sometimes able to reproduce the Qi Gong state but not always. Using the Qi Gong state thus obtained, it was found that this type of Qi Gong energy is directed to specific directions from the hand and can even penetrate wooden or metal doors.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2568075 TI - Effects of the infrared laser therapy at treated and non-treated trigger points. AB - For reliability of the pain threshold measurement there were measured first 390 trigger points of 22 healthy students twice at each point. The reliability of two different measurements was found to be perfect. Infrared (904 nm) laser therapy was compared to placebo laser at the trigger points. Our study tested eighteen patients (11 men and 7 women), with 31 active trigger points in the muscles of the infraspinatus, extensor carpi radialis, levator scapulae, trapezius and tibialis anterior. Trigger points were randomly managed by infrared laser (dose 1.5J/point and place laser. The study was carried out by double-blind and cross over principle. The responses of the management were documented by the pain threshold meter measurements of these trigger points before and after the treatments, and then fifteen minutes later. The trigger points of the other side of the body were also measured from the same muscles. In the results there were observed highly significant changes between the laser and placebo groups immediately after the treatment, 0.97 (SE 0.16) kg/cm2 (p less than 0.001). The differences between these two treatments were greater after fifteen minutes of the therapy--1.87 (SE 0.30) kg/cm2 (p less than 0.001). At the non-treated trigger points, the significant increase of the values was seen after fifteen minutes (p less than 0.05). Our research study results suggest that infrared laser had an effect at the trigger points and that the treatment significantly increased the pain threshold. PMID- 2568076 TI - [A rare hand injury]. AB - Simultaneous dislocations of the carpometacarpal joints are relatively rare hand injuries. This is a case report of dislocations of the carpometacarpal joints II V, combined with Bennett's fracture. The case is discussed in this article with particular reference to the therapeutical aspects. PMID- 2568077 TI - [Isolated fracture of the os trapezium with concomitant dislocation of the carpometacarpal joint of the thumb. A case report]. AB - Fractures of the greater multangular occur mostly combined with additional bone injuries but rarely isolated. This is a report on an isolated fracture of the greater multangular with luxation of the carpometacarpal joint, treated via osteosynthesis. According to most authors and the good result of own case operative treatment is necessary in all dislocated fractures or fractures that cannot be repositioned, to preserve patients from painful arthrosis and reduced function of the thumb. PMID- 2568078 TI - [Fracture of the mandible in osteogenesis imperfecta]. AB - This article is a report on a fracture of the lower jaw in osteogenesis imperfecta. In this disease, fractures of the facial part of the skull are very rare, in contrast to fractures of the extremities; they may possibly occur in adults only if the bone is weakened by additional processes--in the present case, by an odontogenous cyst. In the case described here, successful treatment of the fracture could be effected via intraoral approach by means of a functionally stable osteosynthesis performed with plates. PMID- 2568079 TI - [Fatal penetrating injuries caused by incorrect use of angular abrasive wheels]. AB - Two workmen suffered avoidable fatal injuries from broken parts of abrasive wheels during inexpert handling of these machine tools. The high kinetic energy of these broken machine parts can exercise an effect like bullets. PMID- 2568080 TI - [Use of Biofix C for stabilizing medial ankle fractures]. AB - This report deals with the surgical technique and the results of stabilisation of 10 medial ankle fractures using Biofix C rods. They are cylindrical, biodegredable rods. Biorod is constructed of self-reinforced composite material. It consists of polyglycolide matrix with polyglcolide fibers. The bending strength of the material before operation is more than 200 MPa, exceeding 20-30 times the strength of cancellous bone. The material looses its mechanical strength in hydrolitic conditions during 5-6 weeks and dissolves totally during the following months. Our experience with a self developed guiding tool for the implantation of the Biorod is presented. All fractures healed without any notable complications. As of the 8th day full weight bearing was allowed in a below knee plaster cast for 6 weeks. PMID- 2568081 TI - [Traumatology of sailing injuries]. AB - In a period of 3 years 68 injuries during sailing have been registered. The hand (32.3 p.c.) and the head (17.6 p.c.) are most frequently damaged. Bleeding wounds (26.4 p.c.), contusions (22.0 p.c.) and fractures (17.6 p.c.) are very common lesions. The injuries are mainly caused by the dolorous contact with the boom, by stumbling on board a ship and by jumping on the landing-stage. An effective prevention is discussed. PMID- 2568082 TI - [Surgical complications following implantation of femur head prostheses in a traumatology patient sample. Prevention and therapy]. AB - At the 1st University-Clinic of Traumatology, Vienna, 420 patients with a medial fracture of the femoral neck received a femoral neck prosthesis (59 male, 361 female, phi age 81.3 years; range: 60-97 years). Severe early complications were fractures of the femoral shaft in 1.19%, that healed without problems after cerclage and plate osteosynthesis, and early infections in 3.1%. The rate of infections can be reduced to 1% prophylactic administration of antibiotics, surgery in an laminar airflow operating theater, and by the use of cement containing antibiotics. Dislocation of the prosthesis was observed in 2 patients (0.47%). the low rate of dislocation appears to be due to the antero-lateral operative approach according to Watson-Jones without osteotomy of the trochanter as well as to the precise technique of implantation of the prosthesis. More frequent late complications were periarticular calcification in 20 cases (4.7%) and protrusion in 18 cases (4.28%). However, only one out of the patients with calcification is restricted in his movements, and only 3 out of the patients with protrusion do complain of symptoms. The low rate of symptomatic protrusions is explained by a correct level of resection, by the implantation of a head prosthesis with appropriate dimensions and by the exclusion of patients with arthrosis of the hip or severe osteoporosis from the implantation of a femoral head prosthesis. PMID- 2568083 TI - [Surgical stabilization of femoral fracture following implantation of straight shaft prostheses]. AB - Fractures of the shaft of the femur at the level of a hip endoprosthesis may cause serious problems in regard to operative stabilization as well as to bony union. The therapeutic procedure and the clinical course are described in two patients. The technical procedure is demonstrated on a plastic femur model. The advantages of cementless implantation of the femoral shaft are discussed in view of this complication. PMID- 2568084 TI - [Course of compression-induced compartment syndrome in a drug-dependent patient. A case report]. PMID- 2568085 TI - [Determining indications and tactics in managing the severely injured hand]. AB - The repair of a severely damaged hand requires not only a surgeon's skillful knowledge of the functional anatomy of the hand and the importance of its various parts and their place in the total function, but also experience in microsurgery. One's goal is to extensively save functionally important structures. Besides stabilizing the bone structure, microvascular measures to maintain blood circulation have priority. On the basis of three case studies each with different accident mechanisms, the problems of repairing a severely damaged hand and the results are described. When diagnosing such a severely damaged hand, one must evaluate the functional results that can be expected after repairing the hand in order to decide whether to keep it or to amputate and provide a prosthesis. PMID- 2568087 TI - The epidemiology of testicular cancer in upstate New York. AB - In a study of 250 cases of cancer of the testis and of neighborhood- and age matched controls in upstate New York in 1977-1980, the authors found that risk was enhanced by possession of a number of traits associated with exposure of the testis to heat; occupational exposures to fertilizers, phenols, and fumes or smoke; and trauma to the testis. Risk was also increased for characteristics related to congenital and developmental aberrancies and testis-related abnormalities, e.g., low sperm count, fertility problems, atrophic testis, and cryptorchidism. Several of these risk factors were statistically significant in a multiple regression model that adjusted for all other significant traits, age, and education. PMID- 2568088 TI - Positional effect of cis/trans alpha globin gene deletions on the formation of "H" bodies. AB - Normal individuals have four alpha-globin genes, two on each member of the chromosome 16 pair (alpha alpha/alpha alpha). The alpha-thalassemia trait phenotype associated with deletions of two alpha-genes can be either on the same chromosome, the cis type (alpha alpha/--), or on opposite chromosomes, the trans type (alpha-/alpha-). Traditionally, the observation on vitally stained smears of occasional cells containing "H" bodies has been used as an important diagnostic criterion for alpha-thalassemia trait. These "H" bodies are thought to be precipitated beta tetramers because of the presence of excess beta-globin chains. Our study in patients with various alpha-genotypes indicates that normal subjects (alpha alpha/alpha alpha) and patients with silent alpha-thalassemia trait (alpha alpha/alpha-) generally have no "H" bodies. However, patients with the two-gene deletion of the cis type alpha-thalassemia (alpha alpha/--) show the occasional "H" body, and those with Hb "H" disease (alpha-/-- or alpha cs-/--) show many such bodies. On the other hand, patients with two-gene deletion of the trans type (alpha-/alpha-) do not show "H" bodies. The number of "H" bodies found does not appear to correlate directly with the degree of imbalance in alpha- and beta chain production among the various alpha-genotypes examined. The chemical nature of "H" bodies is discussed, and an alternative hypothesis that embryonic zeta chains expressed in the cis type but not in the trans type of alpha-thalassemia are involved in the formation of "H" bodies is proposed. PMID- 2568086 TI - Comparison of nizatidine and cimetidine as once-nightly treatment of acute duodenal ulcer. Nizatidine Multicenter Duodenal Ulcer Study Group. AB - Nizatidine, a new H2-receptor antagonist for the treatment of duodenal ulcer disease, was compared with cimetidine in an 8-wk, randomized, double-blind, multicenter clinical trial. Patients were randomly allocated to receive either nizatidine 300 mg h.s. or cimetidine 800 mg h.s. Patients were treated for 8 wk, regardless of the healing status of their ulcers. An endoscopy was performed at Wk 2, 4, and 8. Healing rates with nizatidine 300 mg h.s. were numerically, but not statistically significantly, superior to those with cimetidine 800 mg h.s. at each treatment period. Ulcer healing rates at Wk 2, 4, and 8 were 41% (78/191), 73% (130/179), and 81% (145/179) for nizatidine and 33% (60/184), 67% (116/174), and 75% (126/168) for cimetidine, respectively. Symptoms of peptic ulcer disease were similarly reduced at each treatment period by nizatidine and cimetidine. Patients with healed ulcers at either Wk 2 or Wk 4 were continued on therapy and an endoscopy was performed at Wk 8. Ulcer recurrence occurred in 10% of nizatidine-treated and 19% of cimetidine-treated patients at Wk 8 (p = 0.085). The observation of recurrence of duodenal ulcer while patients were receiving full-dose H2-receptor antagonist therapy has not been reported previously. PMID- 2568089 TI - Y chromosome DNA polymorphisms in two African populations. AB - Y chromosome-specific DNA polymorphisms were detected using probe p49f after restriction with TaqI enzyme on samples coming from two African populations: Bantus and Pygmies. All the main TaqI alleles at five Y loci already found in Caucasians are also found in these two populations; 12 of the 16 Caucasian haplotypes were found in these two African populations, and two new haplotypes are Pygmy specific. A proposed phylogeny of the various haplotypes that was derived by using the parsimony criterion established that haplotypes XIII and XVIII, respectively the most frequent one and only one present in Pygmies, are probably ancestral. PMID- 2568090 TI - Fine-structure genetic mapping of human chromosomes using the polymerase chain reaction on single sperm: experimental design considerations. AB - The polymerase chain reaction (PCR) makes it possible to rapidly generate a very large number of copies of a specific region of DNA. Application of PCR to individual human sperm cells permits the typing of a large number of independent male meiotic events. If the donor male is heterozygous at three loci, sperm typing using PCR will permit ordering of loci in a manner analogous to classical methods of experimental genetics. Sequential analysis of trios of loci by sperm typing will provide a very accurate means of ordering any number of tightly linked loci. Here, we describe experimental design and sample-size issues raised by the application of sperm typing by PCR for mapping human chromosomes, and we demonstrate that sperm typing will be an efficient method for generating fine structure human genetic maps. PMID- 2568091 TI - Brother/sister pairs affected with early-onset, progressive muscular dystrophy: molecular studies reveal etiologic heterogeneity. AB - An autosomal recessive (AR) form of muscular dystrophy that clinically resembles Duchenne/Becker types exists, but its frequency is unknown. We have studied three unrelated affected brother/sister pairs and their families for deletions and polymorphisms with the entire dystrophin cDNA and other DNA probes from the Xp21 region to test for involvement of the DMD locus. In family 1 a large intragenic deletion was found in the affected male. The affected sister was heterozygous for this deletion, but the mother was not, implying germinal mosaicism. In family 2, no deletion was detected in the affected male. RFLP analysis revealed that the affected male and an unaffected sister shared a complete Xp21 haplotype while the affected sister had inherited a recombinant Xp21 region resulting from a crossover between pERT 87-15 and J-Bir. Only the 5' region of the dystrophin gene was shared with the affected boy. X-inactivation studies using a polymorphism in the 5'-flanking region of the HPRT gene, in conjunction with methylation sensitive enzymes, revealed random X inactivation in the affected girl's leukocytes. In a muscle biopsy from the affected male, the dystrophin protein was present in normal amount and size. Family 3 was informative for four RFLPs detected with dystrophin cDNA probes which span the entire gene. The affected male was found to share the complete dystrophin RFLP haplotype with his unaffected brother, while his affected sister had inherited the other maternal haplotype. It is concluded that the clinical presentation of early-onset, progressive muscular dystrophy in a male and in his karyotypically normal sister can be caused by mutations at different loci. While in family 1 a deletion in the dystrophin gene is responsible, this gene does not appear to be involved in families 2 and 3. PMID- 2568092 TI - Alternatives to lithium for preventive treatment of bipolar disorder. AB - The authors review the research literature on drug treatment for the prevention of recurrences in bipolar disorder, emphasizing the available alternatives to lithium therapy. They discuss the need for alternative treatments and the current status of promising agents. Carbamazepine receives special attention because of its status as the most promising backup treatment for lithium. The authors conclude that despite the extensive literature on carbamazepine, there is strong need for carefully designed, prospective, double-blind studies to establish the efficacy of carbamazepine alone and in combination with lithium as a prophylactic treatment for bipolar disorder. Difficulties in developing and evaluating preventive maintenance are discussed. PMID- 2568093 TI - Informed consent and tardive dyskinesia. AB - To determine whether a formalized informing process transmitted knowledge concerning the risks and benefits of neuroleptic medication, particularly the risk of tardive dyskinesia, to stable schizophrenic outpatients, the authors administered a multiple-choice questionnaire to 21 patients who were read a standardized information form and 27 patients who were not. The mean scores for the informed patients were significantly higher, and the differences between the two groups remained significant at 6-month follow-up. The information process had no adverse effects on frequency of psychiatric admission, noncompliance with medication, or the need for increased antipsychotic medication. PMID- 2568095 TI - Adjunctive buspirone in benzodiazepine treatment of four patients with panic disorder. AB - Four patients with panic disorder whose panic attacks responded to benzodiazepine treatment but who suffered persistent anxiety improved after addition of buspirone. Despite its lack of antipanic effect, buspirone may offer an adjunctive benefit when added to benzodiazepines in panic disorder. PMID- 2568094 TI - Dopamine blockade and clinical response: evidence for two biological subgroups of schizophrenia. AB - Because CNS neuroleptic concentration cannot be directly measured in patients, the relation between clinical response and extent of dopamine receptor blockade is unknown. This relationship is critical in ascertaining whether nonresponse to neuroleptics is the result merely of inadequate CNS drug levels or of more basic biological differences in pathophysiology. Using [18F]N-methylspiroperidol and positron emission tomography, the authors assessed dopamine receptor occupancy in 10 schizophrenic patients before and after treatment with haloperidol. Responders and nonresponders had virtually identical indices of [18F]N-methylspiroperidol uptake after treatment, indicating that failure to respond clinically was not a function of neuroleptic uptake or binding in the CNS. PMID- 2568096 TI - Neuroleptic-induced dystonia. PMID- 2568097 TI - [Gamma-camera scintigraphy of the testes and the concentration of hormones of the hypophyseo-gonadal axis in patients with cryptorchism and infertility]. PMID- 2568098 TI - The effect of acutely administered phenytoin on vecuronium-induced neuromuscular blockade. AB - Phenytoin was administered intravenously in a dose of 10 mg kg to a group of patients in whom steady state neuromuscular blockade had been established with an infusion of vecuronium. A control group of patients were given 0.9% saline instead of phenytoin. Administration of phenytoin produced significant augmentation of neuromuscular blockade (p less than 0.001). The possible mechanism of this effect is discussed. PMID- 2568099 TI - Peri-operative management of severe thyrotoxicosis with esmolol. AB - A case of severe thyrotoxicosis with cardiac decompensation is reported. Use of propranolol was followed by cardiovascular collapse. Esmolol, the beta-1 adrenoceptor blocking agent was successfully used to control the thyrotoxic state. A discussion of possible advantages of esmolol is provided. PMID- 2568100 TI - Laparoscopy and vagal arrest. PMID- 2568101 TI - Isolation and identification of free amino acids as crystalline N-t butyloxycarbonyl, O-phenacyl derivatives. AB - A general and efficient method for the quantitative isolation of free amino acids from natural sources and their identification as crystalline N-t-butyloxycarbonyl amino acid phenacyl esters is described. The applicability of this method is illustrated in the isolation and characterization of major free amino acids from the Mediterranean fruit fly Ceratitis capitata. PMID- 2568102 TI - Biochemical characterization of an autoradiographic method for studying excitatory amino acid receptors using L-[3H]glutamate. AB - A method was developed for radiolabeling excitatory amino acid receptors of rat brain with L-[3H]glutamate. Effective labeling of glutamate receptors in slide mounted 10-microns sections was obtained using a low incubation volume (0.15 ml) and rapid washing: a procedure where high ligand concentrations were achieved with minimal waste. Saturation experiments using [3H]glutamate revealed a single binding site of micromolar affinity. The Bmax was trebled in the presence of Ca2+ (2.5 mM) and Cl- (20 mM) with no change in the Kd. Binding was rapid, saturable, stereospecific, and sensitive to glutamate receptor agonists. The proportions of [3H]glutamate binding sensitive to N-methyl-D-aspartate (NMDA), kainate, and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) were 34, 54, and 51%, respectively. NMDA inhibited binding at a distinct subset of L-[3H]glutamate sites, whereas AMPA and kainate competed for some common sites. Labeling of sections with L-[3H]glutamate in the presence of the selective agonists allowed autoradiographic visualization of glutamate receptor subtypes in brain tissue. PMID- 2568104 TI - Differing effect of agonist and antagonist muscle relaxants on cat jaw muscles. AB - Mouth closure and an increased resistance to mouth opening follow succinylcholine administration in humans. To elucidate the effects of succinylcholine on masticatory muscle function, mouth opening in the cat, produced by a constant test force, was measured during steady state halothane anesthesia. After baseline measurements, either succinylcholine (0.3 mg.kg-1 of body weight) or vecuronium (0.1 mg.kg-1 of body weight) was infused intravenously, and mouth opening measurements were repeated for up to 30 min. Concomitantly, muscle relaxant effect was quantified by measurement of the neurally-evoked tibialis anterior muscle response. All animals given succinylcholine displayed active jaw closure, which was followed by an increased resistance to mouth opening. This increased resistance was present after cessation of fasciculations and during complete twitch suppression. It lasted beyond the time at which the limb muscle twitch had fully recovered. Vecuronium administration was associated with a decreased resistance to mouth opening without a closing action. The initial jaw closure and the subsequently increased resistance to mouth opening after succinylcholine administration during halothane anesthesia in the cat are comparable with mouth opening changes after succinylcholine administration during inhalation anesthesia in humans. The cat may serve as an animal model for study of the mechanisms involved in responses of jaw muscles to succinylcholine with use of techniques inappropriate in humans. PMID- 2568103 TI - Negative chronotropic actions of sufentanil and vecuronium in chronically instrumented dogs pretreated with propranolol and/or diltiazem. AB - Patients with ischemic heart disease who present for surgery are frequently managed with a "high dose" narcotic technique utilizing fentanyl or sufentanil in combination with neuromuscular blockade. Narcotic anesthetic induction has been associated with perioperative cardiac conduction disturbances, particularly in patients previously treated with calcium channel and/or beta-adrenergic antagonists. The purpose of the present investigation was to examine in chronically instrumented dogs the effects on systemic and coronary hemodynamics and regional contractile function of sufentanil alone or in combination with the nondepolarizing neuromuscular blocking agent vecuronium, with and without pretreatment with the calcium channel blocking agent diltiazem, and/or the beta adrenergic antagonist propranolol. Thirty-six experiments were conducted in four groups of 19 chronically instrumented dogs. Administration of sufentanil (25 and 50 micrograms/kg of normal body weight IV) resulted in a significant sinus or junctional bradyarrhythmia without alteration in systemic and coronary hemodynamics. Pretreatment with propranolol (1 mg/kg IV) decreased left ventricular +dP/dt without other hemodynamic alterations. After propranolol pretreatment, sufentanil (25 and 50 micrograms/kg IV) produced statistically significant dose-dependent decreases in heart rate and +dP/dt, while increasing mean and diastolic coronary vascular resistances. After pretreatment with diltiazem (0.3 mg/kg IV) and propranolol (1.0 mg/kg IV), sufentanil (25 micrograms/kg IV) produced significantly greater bradycardia (38 +/- 4 beats/min) than it did in non-pretreated or propranolol-pretreatment dogs. Similarly, administration of sufentanil (25 micrograms/kg IV) in combination with vecuronium (0.1 mg/kg IV) after propranolol and diltiazem pretreatment resulted in the appearance of bradyarrhythmias (32 +/- 2 beats/min) and two episodes of asystole, responsive to atropine sulfate (0.4 mg). In spite of severe bradycardia, arterial pressure was well maintained in all groups. In all groups, administration of naloxone (0.2 mg/kg IV and 0.2 mg/kg IM) after sufentanil produced significant increases in arterial pressure, left ventricular +dP/dt, and diastolic coronary blood flow velocity. Therefore, the combination of sufentanil and vecuronium in chronically instrumented dogs may result in the appearance of cardiac conduction disturbances. This action is more likely to occur after pretreatment with diltiazem and propranolol. Although systemic hemodynamics were well maintained in dogs in the present study during the episodes of bradyarrhythmias, potentially significant cardiovascular deficits may occur in pa PMID- 2568106 TI - [Optimization of the determination in urine of alanine aminopeptidase, gamma glutamyltransferase and N acetyl-beta-D-glucosaminidase]. AB - The authors describe the optimization of determination of alanine aminopeptidase (AAP), gamma-glutamyltransferase (GGT) and N-acetyl-beta-D-glucosaminidase (NAG) in urine by multivariate analysis. The optimal conditions found are: for AAP at 30 degrees C TRIS HCl buffer 300 mmol/l pH 7.9, L-alanine-4-nitroanilide 5.8 mmol/l, for GGT at 30 degrees C buffer glycylglycine 150 mmol/l pH 8.0, gamma-L glutamyl-3-carboxy-4-nitroanilide 9.0 mmol/l, for NAG at 37 degrees C citrate buffer 50 mmol/l pH 5.8, m cresolsulfonphtaleinyl-N-acetyl-beta-D-glucosaminide 5.5 mmol/l. These methods are easy to perform, apply to urine without pretreatment through Sephadex: therefore complete automatization is possible. The stability of enzymatic activities in urine is of ten days at +4 degrees C in the presence of sodium azide at neutral pH. Freezing resulted in a considerable loss of activity for AAP and GGT. PMID- 2568105 TI - The importance of inhalation of beta 2 sympathomimetic drugs in the home care treatment of asthma in children. AB - In order to improve the care of asthma attacks in children and to reduce hospitalization, 50 patients and their parents were taught a home program using beta 2 sympathomimetic drugs administered by aerosol inhaler. The parents obtained recordings of respiratory rate and pulmonary flow meter. A control group of 50 patients received only slow-release theophylline. Inhalation therapy improved the respiration recorded by reduced respiratory rate and pulmonary flow meter while visits to the emergency department and hospital admissions were reduced by 60%. Absence from school was also reduced by 60%. Benefits to the patient and to cost containment have been achieved by home care treatment. PMID- 2568107 TI - [Use of the recombinant DNA technic in studying cystic fibrosis in 14 Spanish families: detection of carriers and healthy subjects]. AB - Fourteen Spanish families, containing at least one affected child with cystic fibrosis, were typed for restriction fragment length polymorphisms (RFLPs) by proper pJ3.11, pmet H and pmet D. Nine (64.3%) were fully informative for prenatal diagnosis and carrier detection; four (28.5%) were partially informative and prenatal exclusion of an affected fetus could be carried out in half of pregnancies. One (7.1%) was uninformative for these probes. Allelic frequencies obtained have also been analized, being pJ3.11 probe the most informative in our families. PMID- 2568108 TI - The N-type Ca channel in frog sympathetic neurons and its role in alpha adrenergic modulation of transmitter release. PMID- 2568109 TI - Multiple types of calcium channels in heart muscle and neurons. Modulation by drugs and neurotransmitters. PMID- 2568110 TI - Leucopenia during sulphasalazine treatment for rheumatoid arthritis. AB - Leucopenia appears to be a more frequent complication of sulphasalazine treatment in rheumatoid arthritis than in inflammatory bowel disease and poses a management problem. In this study leucopenia was found in 20 patients, 14 of whom were participating in prospective studies (252 patients), giving an incidence of 5.6%. Treatment had to be discontinued in half of these patients. Most (14) episodes of leucopenia occurred early in treatment (less than 24 weeks) but some occurred late and sustained monitoring seems necessary. No predictive factors for leucopenia were elucidated. All patients recovered fully with dose adjustment or, in more severe cases, after withdrawal of sulphasalazine and appropriate supportive therapy. The incidence of leucopenia may be higher in Glasgow than in other units in Britain. PMID- 2568111 TI - Central nervous system infections in the elderly. AB - Review of records of patients aged 65 years and older admitted to The Mount Sinai Hospital, New York, NY, during the period from 1970 through 1985 revealed 57 episodes of central nervous system infections, including 50 meningitides, 5 brain abscesses, 1 subdural empyema, and 1 epidural abscess. Predisposing conditions were present in 17 patients with meningitis, and concurrent infections occurred in 19 patients. Streptococcus pneumoniae accounted for 43% of all isolates; 25% were gram-negative organisms. Of the patients in this sample, fever was present in 100%, meningismus was present in 58%, and change in mental status was present in 86%. Sixty-five percent of patients with meningitis survived; increased mortality was associated with altered mental status, inappropriate initial antibiotic therapy, and hypoglycorrhachia. Delay in diagnosis, underlying disease, and bacteremia did not significantly alter outcome. All patients with focal infections presented with localizing signs and all survived. PMID- 2568112 TI - [Role of plasma histamine and neutrophil chemotactic factor in exercise-induced asthma]. AB - The mechanism of exercise-induced asthma (EIA) is still controversial, although the role of chemical mediator is strongly suspected. In the present study, 50 asthmatic patients were observed after exercise on bicycle ergometer during dry air breathing, and changes of plasma histamine and neutrophil chemotactic factor (NCF) were measured and effect of anti-allergic drugs was examined. 31 patients developed postexertional bronchoconstriction and their % reduction of FEV1 after exercise correlated significantly with the degree of airway hyperresponsiveness to inhaled methacholine determined by Astograph. Plasma histamine levels were examined in 20 EIA positive cases and 13 EIA negative cases, but no significant changes were observed between pre- and post-histamine levels in either group. On the other hand, NCF was elevated significantly after exercise in both EIA positive and negative cases, but postexertional NCF levels were significantly higher in EIA positive than in EIA negative cases. The relationship between % increase of NCF and the % reduction of FEV1 after exercise was significant (r = 0.472, p less than 0.05). DSCG and Azelastine protected the development of EIA in 14 out on 19 cases and 7 out of 12 cases, respectively. Pretreatment with DSCG significantly reduced the increase of NCF after exercise. These results indicates that one of the chemical mediator, NCF, may play an important role in producing postexertional bronchoconstriction in asthmatic patients. PMID- 2568113 TI - Neurotensin, bradykinin and somatostatin inhibit cAMP production in neuroblastoma N1E115 cells via both pertussis toxin sensitive and insensitive mechanisms. AB - Neurotensin, bradykinin and somatostatin inhibited in a time- and concentration dependent manner prostaglandin E1- or forskolin-stimulated cAMP production in neuroblastoma N1E115 cells. Cell treatment with 1 microgram/ml pertussis toxin for 6 hours reversed the inhibition elicited by peptides after short incubation periods (less than or equal to 1 min) but, in contrast, had no effect after longer incubation periods (greater than or equal to 3 min). Fluoroaluminate also inhibited prostaglandin E1-stimulated cAMP production in N1E115 cells, and this effect was not reversed by pertussis toxin. The 6 hour treatment with pertussis toxin was shown to be sufficient to ADP ribosylate virtually all of the 41 kD protein substrate corresponding to the alpha subunit of Gi. Protein kinase C activation with phorbol ester did not inhibit basal or stimulated cAMP production. Our data point to the existence of both pertussis toxin sensitive and insensitive mechanisms of neuropeptide-mediated inhibition of cAMP formation in N1E115 cells. The toxin insensitive response is not mediated by protein kinase C. The possibility is discussed that it results from the activation of a pertussis toxin insensitive G protein. PMID- 2568114 TI - Vitamin A and glutathione-mediated free radical damage: competing reactions with polyunsaturated fatty acids and vitamin C. AB - Vitamin A (retinol reacts extremely rapidly (k = 1.4 x 10(9) M-1 s-1) with thiyl free radicals derived from glutathione to form a free radical with a very strong visible absorption (lambda max. = 380 nm, E max. = 4.0 x 10(4) M-1 cm-1). Arachidonate, linolenate, linoleate and ascorbate also react readily but much more slowly (k = 2.2 x 10(7), 1.9 x 10(7), 1.3 x 10(7) and 3.6 x 10(8) M-1 s-1 respectively). These results support the possibility that vitamin A might play a role in protecting lipid membranes against thiyl free radical mediated damage. PMID- 2568115 TI - Modulation of rapid eye movement sleep in humans by drugs that modify monoaminergic and purinergic transmission. AB - Modulation of rapid eye movement (REM) sleep is a well-established effect of many centrally acting drugs. However, there is uncertainty concerning the nature of the changes and their significance, and it is in this context that we have analyzed the effects of several groups of drugs that alter monoaminergic or purinergic transmission on sleep in humans. The analysis shows that drugs that modulate noradrenergic and serotonergic transmission lead to marked suppression of REM sleep, irrespective of any increase or decrease in sleep duration. There is no evidence that the timing of the ultradian cycle of REM sleep relative to sleep onset is altered by these drugs. On the other hand, reduced REM sleep with dopamimetic drugs is due solely to increased wakefulness. However, there can be more subtle effects of some drugs on REM sleep. Benzodiazepine receptor agonists and drugs that modify purinergic transmission modulate the appearance of early REM activity. There may, therefore, be two discrete systems that control entry into REM sleep, and that are responsive to drugs. The exact appearance and timing of REM periods may be modulated by a feedback mechanism involving GABAergic, or possibly purinergic, transmission, while monoaminergic and cholinergic influences exert a reciprocal and overriding control of REM sleep. PMID- 2568116 TI - Dual aminergic regulation of central beta adrenoceptors. Effect of "atypical" antidepressants and 5-hydroxytryptophan. AB - Nonlinear regression analysis of agonist competition binding curves reveals that the [3H]-dihydroalprenolol-labeled receptor population with low affinity for isoproterenol is increased by p-chlorophenylalanine (PCPA) and this increase is abolished by 5-hydroxytryptophan (5-HTP) in vivo. Desipramine (DMI) decreased the beta adrenoceptor population with high agonist affinity to the same degree in PCPA-treated animals as in control animals, thus explaining the reported discrepancy between beta adrenoceptor number and responsiveness of the beta adrenoceptor-coupled adenylate cyclase system. Mianserin also selectively reduced the beta adrenoceptor population with high agonist affinity in membrane preparations of normal animals, whereas fluoxetine selectively abolished the upregulation of the low affinity sites in reserpinized animals and had no effect on either receptor population from brain of normal animals. The results emphasize the importance of nonlinear regression analysis of agonist competition binding for the interpretation of drug action and encourage the pursuit of the molecular neurobiology of the serotonin (5-HT)/norepinephrine (NE) link in brain. PMID- 2568117 TI - Reversible anorexia and rapid weight loss associated with neuroleptic administration in Alzheimer's disease. AB - Weight loss and malnutrition despite adequate dietary intake has been reported to be part of the clinical course in advanced dementia of the Alzheimer type. We present a case of reversible weight loss associated with neuroleptic use in a patient with Alzheimer's disease and discuss a possible pathophysiological basis for the weight loss. PMID- 2568118 TI - Creutzfeldt-Jakob disease and scrapie prions. AB - Creutzfeldt-Jakob disease, kuru, and Gerstmann-Straussler syndrome are transmissible degenerative diseases of the central nervous system caused by novel infectious pathogens designated prions. Scrapie is a neurodegenerative disease of sheep and goats and is also caused by prions. Experimental scrapie has been extensively studied in hamsters and mice. The scrapie prion protein (PrPSc) is the only component of the infectious scrapie prion identified, to date. Scrapie infectivity and PrPSc copartition into membranes, rods, and liposomes raising the possibility that only PrPSc might be required for infection; however, a second component such as a small nucleic acid cannot be eliminated. PrPSc is encoded by a single copy cellular gene and not by a hypothetical nucleic acid within purified prion preparations. Normal, uninfected cells express the cellular prion protein (PrPc). Both PrPSc and PrPc appear to be translated from the same 2.1-kb mRNA. The N-terminal amino acid sequences of hamster PrPC and PrPSc are identical; both correspond to that predicted by the translated prion protein (PrP) gene sequence. While the chemical difference between PrPc and PrPSc remains unknown, the organization of the PrP gene argues that it results from a posttranslational event. Six posttranslational modifications of both PrP isoforms have been identified: (1) cleavage of an N-terminal signal peptide, (2) an intramolecular disulfide bond, (3) an N-linked oligosaccharide attached to Asn 181, (4) a second oligosaccharide attached to Asn 197, (5) cleavage of a C terminal hydrophobic peptide, and (6) a phosphatidylinositol glycolipid attached to the C-terminus. The mouse PrP gene is on chromosome 2 and is linked to a gene controlling the scrapie incubation time (Prn-i). PrP genes from inbred mice with short and long incubation times differ by two amino acids, a finding consistent with but not proving that PrP modulates susceptibility to scrapie. PrPSc stimulation of a posttranslational process which converts PrPc or its precursor into PrPSc is one possible mechanism for prion replication. This is consistent with observations showing that human prion diseases are manifest as infectious, sporadic and genetic disorders. PMID- 2568119 TI - The nature of the unconventional slow infection agents remains a puzzle. AB - Unconventional slow infections are progressive transmissible degenerative disorders of the central nervous system. The human diseases belonging to this group are Creutzfeld-Jakob disease, kuru, and Gerstmann-Straussler syndrome. Scrapie, transmissible mink encephalopathy, chronic wasting disease of mule deer and elk, and the recently discovered bovine spongiform encephalopathy are similar diseases found in animals. Unusual characteristics of the unconventional slow infections clearly distinguish these disorders from conventional infections. These include: unusually long incubation periods (from months to years); progressive CNS degeneration with characteristic histopathological lesions; the lack of an immune or inflammatory response; unconventional biological and physical properties of the etiologic agents. There has been considerable controversy concerning the nature of the causative agent. The 3 main hypotheses, virus, virino, and modified host protein, are reviewed relative to their ability to explain the properties of the agent and the unusual characteristics of the disease process. The discovery of an abnormal structure, termed scrapie associated fibrils (SAF) and an abnormally modified 33-37 kDa host-encoded glycoprotein unique to unconventional slow infections opened new areas of intense interest and investigation. SAF are abnormal filamentous structures which copurify with infectivity and possess characteristics of "amyloids." The major component of SAF is the host-encoded scrapie-specific protease resistant glyco protein. Considerable data has accumulated on the biochemistry, immunology and molecular biology of this host coded scrapie protein. The relationship of SAF and the scrapie-specific protein to the infectious agent is discussed in the context of each of the "nature of the agent" hypotheses. PMID- 2568120 TI - Multiple endocrine neoplasia type 1 (MEN-1). Clinical, biochemical and genetical investigations. AB - The syndrome of multiple endocrine neoplasia type 1 is an autosomal dominantly inherited disease affecting several endocrine organs. The affected organs include the pituitary, the parathyroids and endocrine pancreas, where different types of lesions can be found, such as hyperplasia or frank carcinomas. The most life threatening lesions are the endocrine pancreatic tumors, which cause about 80% of all deaths among the MEN-1 members. In our own series of 108 members from 16 families with multiple endocrine neoplasia, 55 members had the MEN-1 trait. Among these members, pituitary lesions were found in 42%, parathyroid involvement in 89% and endocrine pancreatic tumors in 58%. Hyperparathyroidism was the presenting lesion of the MEN-1 trait. By using a specific meal stimulation test we have been able to unveil pancreatic lesions up to a median of five years previous to radiological detection. Very recently we have been able to detect a specific genetic lesion in MEN-1 members by studying DNA rearrangements with recombinant DNA technique, using the method of polymorphic restriction enzyme recognition in three large kindreds. The MEN-1 locus maps to chromosome 11q and the MEN-1 predisposition would be a constitutional mutation in heterozygous form, inherited as an autosomal dominant trait. Tumor development involves a second mutational event which involves the chromosome 11, carrying the remaining 'wild' type allele at the MEN-1 locus by means of chromosome loss event. Survival analysis demonstrates that patients with the MEN-1 syndrome had a significantly better survival from diagnosis than patients with sporadic endocrine pancreatic tumors (median 15.1 years and 5.8 years respectively, p = 0.0068). Earlier diagnosis and start of treatment might account for a longer survival in the MEN-1 group, but a possibility of differences in tumor biology between familial and sporadic endocrine pancreatic tumors cannot be ruled out. The surgical treatment of patients with MEN-1 include resection of the parathyroids with transplantation of a piece of the gland to the forearm, resection of endocrine pancreatic tumors in the tail and local enucleation of tumors in the pancreatic head and body. Total pancreatectomy should be avoided in most instances. The causative medical treatment of patients with malignant endocrine pancreatic tumors and the MEN-1 trait include chemotherapy (streptozotocin plus 5-fluorouracil), interferons and the somatostatin analogue SMS 201-995. PMID- 2568121 TI - Treatment of the pancreatic disease of multiple endocrine neoplasia type 1 (MEN 1). AB - This review comprises an analysis of the treatment alternatives for pancreatic tumours of the MEN 1 syndrome presently in use at the Surgical Endocrine Unit of Sahlgrenska Hospital, Goteborg, Sweden. Patients with pancreatic tumours localized by imaging studies are offered surgical excision. Otherwise the therapy is individualized. Some patients with Zollinger-Ellison syndromes are managed with medical antiulcer treatment, others are treated with total gastrectomy. MEN 1 patients with other syndromes, e.g. insulinomas, glucagonomas and VIPomas, are offered surgery with the aim of tumour excision. Even noncurative excision might benefit the management of these cases and possibly prolong life. Asymptomatic patients with raised hormonal tumour markers in whom imaging studies have failed to reveal any tumours, are continuously followed without treatment. The malignant behaviour of the pancreatic tumours varies between different MEN 1 families. MEN 1 patients are considered for total pancreatectomy if they have multiple malignant pancreatic tumours and if the pancreatic disease has caused high morbidity and mortality in the family. PMID- 2568122 TI - Glucocorticoid receptor regulation: the effects of adrenalectomy, exogenous glucocorticoid, and stress on hepatic receptor number in male and female mice. AB - Although glucocorticoids are known to regulate their own receptor number, the physiologic significance of this process is not known. In order to assess this process in intact animals the effects of adrenalectomy, stress, and exogenous glucocorticoid on the number of hepatic glucocorticoid receptors in Swiss-Webster mice were evaluated. In males 24 hr after adrenalectomy there was a clear 2- to 2.5-fold increase (upregulation) in glucocorticoid receptor number. Conversely, 24 hr after the ip administration of exogenous corticosterone there was a clear downregulation of receptor number. In each case (upregulation and downregulation) female mice were much less responsive than males. Three stressors were used to evaluate the effect of the endogenous secretion of glucocorticoids on downregulation. Male mice were exposed to ether, vibration, and confinement either once or daily for periods up to 3 days. Animals were sacrificed 24 hr after the last stress and hepatic receptor number was compared to an unstressed control. Cytosolic receptor number was not influenced by any of these stimuli. It is concluded that although glucocorticoids clearly regulate glucocorticoid receptor number, as demonstrated by adrenalectomy and the administration of steroids to adrenalectomized animals, the physiologic significance of this process is uncertain as receptor number does not appear to be changed by stimuli of adrenal glucocorticoids in the intact animal. PMID- 2568124 TI - Substrate variability as a factor in enzyme inhibitor design: inhibition of ovine brain glutamine synthetase by alpha- and gamma-substituted phosphinothricins. AB - Ovine brain glutamine synthetase (GS) utilizes various substituted glutamic acids as substrates. We have used this information to design alpha- and gamma substituted analogues of phosphinothricin [L-2-amino-4 (hydroxymethylphosphinyl)butanoic acid], a naturally occurring inhibitor of GS. These compounds display competitive inhibition of GS, and a correlation between the inhibitor Ki values and the Km/Vmax values of the analogously substituted glutamates supports the hypothesis that the phosphinothricins participate in transition-state analogue inhibition of GS. At concentrations greater than Ki these inhibitors caused biphasic time-dependent loss of enzyme activity, with initial pseudo-first-order behavior; k'inact parameters were determined for several compounds and were similar to the 2.1 X 10(-2)s-1 value measured for PPT. Dilution after GS inactivation caused a non-first-order recovery of activity. Reactivation kinetics were insensitive to inhibitor and ADP concentrations over wide ranges, although very high postdilution concentrations of inhibitor suppressed reactivation. The burst activity level, beta, as well as the concentration of inhibitor required to suppress reactivation to this level, mu, expressed as a multiple of the Ki value, was characteristic for each compound in the phosphinothricin series. Increasing substitution of the phosphinothricin parent structure caused an increase in Ki values as well as in the inactivation/reactivation parameters. The kinetic behavior of these inhibitors is consistent with a mechanistic scheme involving initial phosphorylation and rapid partial inhibitor dissociation, followed by slow release of remaining bound inhibitor. PMID- 2568123 TI - Requirement for proliferating cell nuclear antigen expression during stages of the Chinese hamster ovary cell cycle. AB - Proliferating cell nuclear antigen (PCNA/cyclin) is a nuclear protein that can stimulate purified DNA polymerase delta in vitro, and its synthesis correlates with the proliferation rate of cells. We have attempted to determine whether synthesis of PCNA/cyclin in Chinese hamster ovary cells is necessary to regulate entry into S phase. We have measured cellular PCNA/cyclin concentration of the mRNA or protein throughout the cell cycle. Cells were separated by centrifugal elutriation into populations enriched for G-1, S, and G-2/M phases. Quantitative Northern hybridization analysis was performed on RNA isolated from each cell population by using a cDNA clone of PCNA/cyclin as a probe. Results demonstrated that although intact PCNA/cyclin mRNA is present during all phases of the cell cycle, an induction of about 3-fold occurs during S phase. Two-parameter staining for PCNA/cyclin and DNA, and analysis by flow cytometry, confirmed that the quantity of PCNA/cyclin protein in the cells increases severalfold in G-1 or early S phase but generally is invariant in S and G-2/M phases. This cell cycle dependence of PCNA/cyclin expression suggests that the observed synthesis is a prerequisite for initiation of DNA replication. Introduction of an antisense oligonucleotide complementary to the PCNA/cyclin mRNA to inhibit PCNA/cyclin synthesis effectively prevented entry of G-1 phase cells into S phase. A complementary sense oligonucleotide used as a control did not have an inhibitory effect. This result suggests that a threshold concentration of PCNA/cyclin is necessary for entry into S phase. PMID- 2568125 TI - Interaction of anions and ATP with the coated vesicle proton pump. AB - ATP-driven proton transport in intact clathrin-coated vesicles requires the presence of a permeant anion, such as Cl-, to provide charge compensation during the electrogenic movement of protons. Using the purified (H+)-ATPase from clathrin-coated vesicles in both the detergent-solubilized and reconstituted states, we have studied the direct effects of anions on the activity of this enzyme. Both proton transport and ATP hydrolysis by the purified enzyme are independent of the presence of Cl-. In addition, proton transport does not occur even at high Cl- concentrations unless K+ and valinomycin are present to dissipate the membrane potential generated. These results indicate that the anion channel which provides for Cl- flux in intact coated vesicles is not a component of the purified (H+)-ATPase. Inhibition of ATPase activity is observed in the presence of I-, NO3-, or SO4(2-), with 50% inhibition occurring at 350 mM I-, 50 mM NO3-, or 40 mM SO4(2-). The presence of ATP lowers the concentration of I- required for 50% inhibition from 350 mM to 100 mM and increases the maximal inhibition observed in the presence of NO3- from 65% to 100%. Two separate mechanisms appear to be responsible for anion inhibition of the (H+)-ATPase. Thus, I- and high concentrations of NO3- (in the presence of ATP) cause inhibition by dissociation of the (H+)-ATPase complex, while SO4(2-) and NO3- (in the absence of ATP) cause inhibition without dissociation of the complex, suggesting the existence of an inhibitory anion binding site on the enzyme.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568126 TI - Isolation, characterization, and amino acid sequence of a polypeptide neurotoxin occurring in the sea anemone Stichodactyla helianthus. AB - An aqueous exudate collected from frozen and thawed bodies of a Caribbean sea anemone, Stichodactyla (formerly Stoichactis) helianthus, contained a polypeptide neurotoxin (Sh I) selectively toxic to crustaceans. The polypeptide was purified by G-50 Sephadex, phosphocellulose, and sulfopropyl-Sephadex chromatography and shown to have a molecular size of 5200 daltons and a pI of 8.3. The amino acid sequence determined by automatic Edman degradations of whole RCM Sh I and of its clostripain, staphylococcal protease, and cyanogen bromide digest peptides is A1ACKC5DDEGP10DIRTA15PLTGT20VDLGS25CNAGW30EKCAS35YYTII40ADCCR45KKK . Only 33% of this sequence is identical with the sequence of Anemonia sulcata toxin II, a sea anemone toxin isolated from the taxonomic family Actiniidae. The six half cystines are located in equivalent positions to those of the actiniid toxins and account for nearly half of the residues common to all of the toxins. However, 69% of the Sh I sequence is identical with that of toxin II from Heteractis paumotensis, another sea anemone belonging to the family Stichodactylidae. Stichodactylid toxins lack the initial N-terminal residue of actiniid toxins and possess three consecutive acidic residues at positions 6-8, a single tryptophan at position 30, and four consecutive basic residues at positions 45-48 (C terminus). A rabbit IgG prepared by Sh I immunization bound Sh I with a K0.5 of 4.7 nM but failed to bind homologous actiniid (Anemonia sulcata II, Condylactis gigantea III) or bolocerid (Bolocera tuedae II) polypeptide neurotoxins.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568128 TI - Inhibition of aminopeptidases by aminophosphonates. AB - More than 30 aminophosphonates were synthesized to probe how the structural changes introduced into the phosphonic acid analogue of leucine, a potent inhibitor of cytosolic leucine aminopeptidase (Giannousis & Bartlett, 1987), affect their ability to inhibit cytosolic (EC 3.4.11.1) and microsomal (EC 3.4.11.2) aminopeptidases. Although most of the compounds studied were found to exert only a modest competitive inhibitory effect, nearly every modification of the structure of the phosphonic acid analogue of leucine was reflected in a marked difference in the affinities of these compounds for the two enzymes. [1 Amino-2-(N-alkylamino)ethyl]phosphonic acids are effective inhibitors of the microsomal enzyme, acting in a time-dependent manner. Kinetic data obtained for these inhibitors correspond to the mechanism for a biphasic slow-binding inhibition process: E + I in equilibrium E* in equilibrium E*I, in which the slow initial isomerization of the enzyme is followed by the fast formation of enzyme inhibitor complex. The most effective inhibitor of this type was [1-amino-2-(N cyclohexylamino)ethyl]phosphonic acid, which has a Ki value of 0.87 microM toward the microsomal aminopeptidase--a value that can be considered as equipotent with bestatin and with leucinal and hydroxamic acids, the strongest known nonpeptide inhibitors of this enzyme. PMID- 2568127 TI - Chloroplast ribosomal protein L12 is encoded in the nucleus: construction and identification of its cDNA clones and nucleotide sequence including the transit peptide. AB - An architectural feature found in all classes of ribosomes is a thin, 10-nm-long protuberance in the large subunit, generated by multiple copies of r-protein L12. The primary structure of spinach chloroplast r-protein L12 is known [Bartsch, M., Kimura, M., & Subramanian, A. R. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 6871 6875], but the location of its gene, whether in the organelle or in the nucleus, has not been determined. Therefore, we synthesized four oligodeoxynucleotides based on the amino acid sequence data and used them to probe a spinach cDNA library we constructed in lambda gt11 vector. cDNA inserts from four of the hybridizing recombinant clones were characterized and sequenced. The data showed that they are reverse transcripts of varying length, all derived from a single poly(A+) RNA species. The longest cDNA molecule is 900 base pairs (bp) long and includes a 5' noncoding sequence followed by two neighboring AUG codons both in the consensus, eukaryotic initiator context, a 56-codon-long transit peptide sequence (starting from the first AUG codon), the amino acid sequence of mature L12 protein, and a 238 bp long 3' downstream noncoding sequence including a polyadenylation signal and the start of the poly(A) tail. The transit peptide sequence has an unusual amino acid composition similar to that of other known chloroplast transit peptides. Northern blot analysis of the poly(A+) RNA isolated from spinach seedlings and probed with the cDNA insert revealed the occurrence of a strong, broad, 950-nucleotide-long band of the corresponding poly(A+) containing mRNA species.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568129 TI - Low affinity of beta-adrenergic receptors for agonists on intact cells is not due to receptor sequestration. AB - The low affinity of beta-adrenergic receptors for agonists described on intact cells at 37 degrees C has usually been interpreted in terms of reduced accessibility of agonists (which are usually hydrophilic) for sequestered receptors. We challenged this hypothesis by eliminating the plasma membrane barrier with low doses of the detergent digitonin. In human mononuclear leukocytes (MNL) permeabilized with digitonin, sequestered receptors became accessible to hydrophilic ligands such as agonists, but the affinity was still low. Then we investigated the relationship between low affinity agonist binding and sequestration using concanavalin A, which blocks sequestration. Even when sequestration was blocked, the affinity of the beta-adrenergic receptors for agonists was low. We conclude that: (a) low affinity agonist binding is independent of receptor sequestration; (b) the receptors which undergo conformational change are those that are sequestered; (c) the low affinity appears before sequestration occurs. This receptor conformational change could be the first step in agonist-induced desensitization. PMID- 2568130 TI - [Dependence of kinetic constants of intact human lymphocyte beta2-adrenoreceptors on the nature of the radioligands used]. AB - An analysis of human peripheral blood intact mononuclear lymphocyte beta 2 adrenoreceptors showed that the hydrophilic radioligand 3H-CGP-12177 binds at various temperatures only to surface receptors. The density of receptors as determined by the binding of the lipophilic antagonist 125I-CYP at 37 degrees C is 2 times as high as that at 4 degrees C. The affinity of lymphocyte beta 2 adrenoceptors for 1-isoproterenol measured by the labeled ligand displacement at low (4 degrees C) temperatures is by two or three orders of magnitude higher than that at 37 degrees C. The thiol-alkylating agent, N-ethylmaleimide (NEM) causes oppositely directed changes in the density of beta 2-adrenoceptors on intact lymphocytes depending on their original density. NEM increases the affinity of beta 2-adrenoreceptors for the hormone (1.5-12-fold). The state and regulation of human peripheral blood lymphocyte beta 2-adrenoreceptors depend on the hormonal status of patients at the moment of blood sample collection. PMID- 2568131 TI - Cholesterol metabolism in infant rats, effect of 6-hydroxydopamine and guanfacine. AB - Infant rats (10-16 days) and weaned animals (older than 18 days) were treated with drugs inhibiting beta-adrenergic activity and were sacrificed 4 days later. 6-Hydroxydopamine acts on postsympathetic nerve fibres, and guanfacine stimulates alpha 2-receptors. Both drugs caused a rise in plasma cholesterol and a decrease in hepatic 3-hydroxy-3-methylglutaryl CoA reductase activity in infant but not in weaned animals. The activity of acylcholesterol acyl CoA transferase, on the other hand, was decreased after drug administration. Thus, both antihypertensive drugs may cause changes in cholesterol metabolism, especially in infant animals. PMID- 2568132 TI - The effect of neuroleptic discontinuation on psychopathology, involuntary movements, and biochemical measures in patients with persistent tardive dyskinesia. AB - As some of the pharmacological activities of neuroleptic medication may involve pathophysiological mechanisms underlying schizophrenia and tardive dyskinesia (TD), it is useful to study patients undergoing medication discontinuation. In this study, 19 stable, neuroleptic-maintained patients with persistent TD underwent taper and discontinuation of their neuroleptic medication over a 3-week period, and multiple behavioral and biochemical (plasma HVA, MHPG, and prolactin) measures were obtained. The major finding was that early relapsing patients had lower baseline and a significantly greater increase in plasma HVA levels after discontinuation than nonrelapsing patients. In addition, patients exhibiting withdrawal-exacerbated TD had significantly lower plasma MHPG levels than patients not exhibiting this phenomenon. The clinical and pharmacological implications of these findings are discussed. PMID- 2568133 TI - Electroconvulsive therapy in neuroleptic-induced parkinsonism. AB - The effect of electroconvulsive therapy (ECT) on the severity of neuroleptic induced parkinsonism was studied in nine schizophrenic inpatients in a longitudinal triphasic design: neuroleptics-neuroleptics plus ECT-neuroleptics. The results suggest that ECT has a true antiparkinsonian potential. The role of ECT in the treatment of Parkinson's disease, especially with therapy-resistant patients complicated with on-off symptoms, is highlighted. PMID- 2568134 TI - Effects of GTP on hormone-stimulated adenylate cyclase activity in cerebral cortex, striatum, and hippocampus from rats treated chronically with lithium. AB - The effects of lithium on guanosine triphosphate (GTP) stimulated adenylate cyclase activity and hormone-induced GTP activation of the enzyme have been studied in three regions of the rat brain. Chronic treatment with lithium, giving a serum lithium level of 0.71 +/- 24 mmol/L, reduced isoprenaline-induced GTP stimulation of adenylate cyclase activity in cortical membranes at concentrations of GTP up to 2 microM. No effect of lithium was observed at higher concentrations of GTP. The enzyme activity stimulated by GTP alone was unaltered by lithium ex vivo. In striatal membranes, lithium ex vivo decreased both dopamine-induced GTP activation of adenylate cyclase and GTP-stimulated adenylate cyclase activity at concentrations of GTP below 2 microM. No effects of lithium ex vivo were found in striatum at 2 microM GTP and above. In hippocampal membranes, lithium ex vivo did not influence either serotonin-induced GTP stimulation of the adenylate cyclase or GTP-stimulated enzyme activity at low levels of GTP. However, at 50 microM GTP, lithium ex vivo enhanced serotonin-stimulated enzyme activity. The present results suggest that lithium ex vivo decreases neurotransmitter activation of the cortical beta-adrenergic adenylate cyclase by influencing the mechanisms by which receptor agonists enhance the GTP stimulation of the adenylate cyclase. Furthermore, lithium ex vivo exerts a region-specific action on the brain adenylate cyclases, but in the brain regions studied, an effect of lithium on N protein level might be of significance for the action of lithium ex vivo on neurotransmitter activation. PMID- 2568135 TI - A mechanism for the involvement of colocalized neuropeptides in the actions of antipsychotic drugs. AB - Evidence has accumulated to implicate neuropeptides localized within midbrain dopamine neurons (cholecystokinin, neurotensin, acetylcholinesterase) in synaptic transmission, mental disease, and pharmacotherapy. We suggest a means by which antipsychotic drugs alter the dynamics between dopamine and colocalized peptides: the intrinsic ability of these agents to stimulate dopamine neuronal activity while blocking dopamine receptors modulates the ratio of catecholaminergic to peptidergic transmission within the mesotelencephalic system. Imbalances of peptide and dopamine cotransmission and their modulation by neuroleptics may be relevant to the pathogenesis and pharmacotherapy of schizophrenia. PMID- 2568136 TI - A self-contained life support system designed for use with a portable hyperbaric chamber. AB - Recently, the Gamow Bag, a portable hyperbaric chamber, has been demonstrated to relieve the symptoms of acute mountain sickness (AMS). The patient is completely enclosed in the bag which is inflated and pressurized to simulate descent in altitude. CO2 produced by the patient is vented from the airtight bag by means of a pressure relief valve, while fresh air is brought in from the outside via a high volume foot pump. In order to eliminate the vigorous pumping that is necessary to maintain a suitable atmosphere in the bag, we have designed a completely portable, self-contained life support system that supplies oxygen as it is consumed and removes the waste CO2 as it is produced. The entire rebreathing unit, which maintains a homeostatic atmosphere in the chamber for six to eight hours weighs less than six pounds. The chamber with the self-contained life support system weights less than 18 pounds. It would find its greatest use in medical mountain clinics, isolated ski areas and should become standard equipment for mountain search and rescue units. PMID- 2568137 TI - Benefit-risk of agonist-antagonist analgesics. PMID- 2568138 TI - Pharmacology of nicotine. AB - In recent years progress in basic neuropsychopharmacology and clinical addiction research have allowed the conclusion that tobacco smoking essentially represents an addiction to nicotine. Parallel to this work, experimental research in biochemistry, physiology and pharmacology has provided detailed descriptions of the structure and function of the nicotinic receptor, the biologic mediator of the many actions of nicotine. This article reviews current knowledge of nicotinic mechanisms in the peripheral and central nervous systems as well as some implications for the notion of smoking as an addiction to nicotine. In particular this review will focus on the effects of nicotine on brain dopamine and noradrenaline systems since these neuronal systems appear to be crucially involved in the rewarding and stimulant effects of addictive drugs. PMID- 2568140 TI - Noradrenaline (gamma) and ATP responses of innervated and non-innervated rat cerebral arteries. AB - 1. The distribution of sympathetic adrenergic nerves on the rat middle cerebral artery and on the arterioles which originated from it was determined by use of gloxylic histochemistry. 2. Whereas the middle cerebral artery and proximal arterioles arising from this artery received a sympathetic innervation, the distal regions of the same arterioles were devoid of innervation. 3. The arteries and arterioles which were innervated were depolarized by noradrenaline in the combined presence of alpha- and beta-adrenoceptor antagonists. Those which were not innervated were not depolarized by noradrenaline. 4. ATP depolarized all arteries and arterioles examined. 5. These observations are discussed with respect to the similarities and differences between gamma-adrenoceptors and P2 purinoceptors. PMID- 2568139 TI - Alpha 1-adrenoceptor antagonist activity of novel pyrimidine derivatives (SHI437 and IK29) in rabbit aorta and trigone of the bladder. AB - 1. In the rabbit isolated aorta and trigone of the bladder, noradrenaline, phenylephrine and clonidine elicited concentration-dependent contractions, which may be caused through activation of postsynaptic alpha 1-adrenoceptors. 2. SHI437, IK29, prazosin and yohimbine competitively antagonized the contractile responses induced by noradrenaline in the aorta and trigone. The pA2 values of SHI437, IK29, prazosin and yohimbine were 7.35 +/- 0.09, 7.47 +/- 0.10, 8.55 +/- 0.02 and 6.28 +/- 0.05 in the aorta, and 8.07 +/- 0.04, 8.30 +/- 0.03, 8.22 +/- 0.04 and 6.46 +/- 0.04 in the trigone, respectively. 3. SHI437, IK29, prazosin and yohimbine also possessed competitive alpha 2-adrenoceptor blocking properties, judging from their antagonism of the clonidine-induced inhibitory effect on the twitch responses in the electrically stimulated vas deferens of the rat. The pA2 values of SHI437, IK29, prazosin and yohimbine were determined to be 4.76 +/- 0.02, 4.74 +/- 0.02, 5.06 +/- 0.03 and 7.86 +/- 0.04, respectively. 4. SHI437, IK29 and prazosin inhibited the contractile responses elicited by transmural electrical stimulation without affecting the evoked 3H-overflow from the [3H]-noradrenaline-preloaded rabbit aorta. Yohimbine augmented the contractile responses and 3H-overflow. 5. SHI437 and IK29 at a concentration sufficient to inhibit noradrenaline-induced contraction failed to attenuate the contractile responses of aorta to KCl, 5-hydroxytryptamine and prostaglandin F2 alpha, and of the trigone to acetylcholine and histamine. 6. The present results suggest that SHI437 and IK29 are highly selective alpha 1-adrenoceptor antagonists, especially in the trigone of the bladder. PMID- 2568141 TI - Potentiation of atrial natriuretic peptide-stimulated cyclic guanosine monophosphate formation by glucocorticoids in cultured rat renal cells. AB - 1. The effect of steroid hormones on atrial natriuretic peptide (ANP)-stimulated cyclic guanosine monophosphate (cyclic GMP) formation was studied in cultured rat renal cells. 2. ANP increased cyclic GMP formation in a dose-dependent manner, while cyclic AMP was not changed by ANP. 3. Steroid hormones did not affect basal cyclic GMP levels in cultured rat renal cells. 4. Dexamethasone at 10(-8) M increased ANP (human and rat ANP)-stimulated cyclic GMP dose-dependently in cultured rat renal cells. Cortisol, corticosterone and aldosterone at a concentration of 10(-7) M also potentiated ANP-stimulated cyclic GMP formation, although triiodothyronine, oestradiol and testosterone were ineffective. Potentiation of ANP action by these steroids seems to parallel glucocorticoid activity. 5. Dexamethasone did not affect cyclic GMP formation stimulated by sodium nitroprusside which stimulates soluble guanylate cyclase in the cytosol. Therefore, the potentiating action of dexamethasone may be mediated through the action on particulate guanylate cyclase at the plasma membrane. 6. It is suggested that the diuretic action of glucocorticoids may, at least in part, be mediated through the potentiating effect of glucocorticoids on cyclic GMP response to ANP. PMID- 2568142 TI - Reversal of the cardiotonic and action-potential prolonging effects of DPI 201 106 by BDF 8784, a methyl-indol derivative. AB - 1. We studied the interaction of the cardiotonic compound DPI 201-106 (4-[3'-(4'' benzhydryl-1''-piperazinyl)-2'-hydroxypropoxy]-1H-indole-2- carbonitrile; DPI) and its derivative BDF 8784 (2-methyl-4-[3'-(4''-benzhydryl- 1''-piperazinyl)-2' hydroxypropoxy]-1H-indole; BDF) in isolated right ventricular papillary muscles of guinea-pig heart. 2. In contrast to the cardiotonic DPI, the methyl-indole derivative lacked a positive inotropic effect and even caused negative inotropic effects in concentrations above 1 microM. At 10 microM BDF significantly reduced the force of contraction and dV/dtmax, but did not affect action potential duration (APD). 3. Pretreatment of papillary muscles with BDF prevented the positive inotropic action of DPI in a concentration-dependent, but non competitive fashion. At 10 microM, BDF prevented the inotropic effect of racemic DPI and shortened the DPI-induced prolongation of action potential duration. BDF similarly affected the inotropic and APD-prolonging effects of the sea anemone polypeptide ATX II. 4. In cardiac myocytes, DPI induced a tetrodotoxin (TTX) sensitive, slowly inactivating inward current. The slow decay of this current was enhanced by BDF. In cells pretreated with BDF, DPI was not effective. BDF alone depressed the sodium and the calcium current. 5. In conclusion, the non-inotropic methyl-indole derivative BDF interacts with DPI noncompetitively at the sodium channels to abolish the inotropic and APD-prolonging effects of DPI, emphasizing the importance of the substituent in position 2 of the indole moiety for this effect. PMID- 2568143 TI - Evidence that central 5-hydroxytryptaminergic neurones are involved in the anxiolytic activity of buspirone. AB - 1. A two-compartment exploratory test was used to assess the role of central 5 hydroxytryptaminergic neurones in the anxiolytic activity of buspirone in rats. 2. Buspirone 0.1 mg kg-1, administered subcutaneously 15 min before testing, significantly increased black-white transitions (BWT) in control rats but had no effect in animals injected intracerebroventricularly one week before with 150 micrograms 5,7-dihydroxytryptamine (in 20 microliters). 3. Infusion of buspirone in the median raphe (but not in the dorsal raphe) significantly enhanced BWT, at doses from 1 micrograms to 10 micrograms (in 0.5 microliters). Buspirone 5 and 10 micrograms, but not 1 microgram, administered in the median raphe, significantly enhanced motor activity of rats during the first 10 min of testing in the activity cages. 4. The effect on BWT of 5 micrograms buspirone in the median raphe was completely antagonized in animals which had received either 5,7 dihydroxytryptamine intraventricularly, 150 micrograms (in 20 microliters), one week before or an infusion of 0.1 microgram (in 0.5 microliter) (-)-propranolol in the same area 5 min before. (-)-Propranolol infused in the median raphe did not modify the effect of buspirone on locomotion. 5. Infusion of 5 micrograms buspirone (in 0.5 microliter) in the median raphe significantly enhanced punished responses in a conflict test with no effect on unpunished responding. Buspirone infused in the dorsal raphe had no effect on punished or unpunished responding over a wide dose range. 6. The results indicate that at the relatively low dose used in the present study buspirone produces an anxiolytic effect by acting on central 5-hydroxytryptaminergic neurones. It is likely that activation of 5 hydroxytryptamine1A-receptors in the median raphe is involved. PMID- 2568144 TI - 8-OH-DPAT, flesinoxan and guanfacine: systemic and regional haemodynamic effects of centrally acting antihypertensive agents in anaesthetized rabbits. AB - 1. 8-Hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) and flesinoxan, agents which show high affinity and selectivity for 5-HT1A receptors, were administered intravenously in doses of 0.003 to 0.1 and 0.01 to 0.3 mg kg-1 respectively to 5 rabbits each. Their effects were compared with those of the centrally acting agent and alpha 2-adrenoceptor agonist, guanfacine, 0.01-0.3 mg kg-1, administered to a group of 5 rabbits. Five further rabbits were used as controls and treated with the vehicle of the active agents. 2. Both flesinoxan and 8-OH DPAT induced similar systemic and regional haemodynamic changes. Both lowered mean arterial pressure and heart rate. The principal blood pressure lowering mechanism was vasodilatation; cardiac output changed minimally despite the falls in heart rate and myocardial contractile force. 3. With guanfacine the maximal fall of blood pressure was comparable to that obtained with the 5-HT1A receptor ligands; however, in contrast to the latter, the dose-response curve was U shaped, the highest dose eliciting a pressor effect with reversal of the vasodilatation. 4. Widespread peripheral vasodilatation was found with all the agents in the splanchnic circulation and also in the brain and skeletal muscle. A weak tendency towards vasodilatation was found in the kidneys where the dose response curve was bell-shaped for guanfacine. 5. This spectrum of activity is different from that of peripheral vasodilators, such as calcium antagonists, potassium channel activating agents or hydralazine; it is, however, consistent with the putative mechanism of action of these compounds to reduce peripheral sympathetic tone by a central mechanism of action. PMID- 2568147 TI - Staphylococcus aureus septicaemia: the initial presentation of polyarteritis nodosa. AB - A 51-year-old man presented with Staphylococcus aureus septicaemia. He improved initially but died suddenly on day 8 of his hospital admission. Post-mortem examination revealed multiple lesions of polyarteritis nodosa which were the cause of death. We are unaware of any other case of polyarteritis nodosa that presented with Staphylococcus aureus septicaemia. These two conditions have a number of similar clinical characteristics and are difficult to distinguish unless the association is considered. PMID- 2568145 TI - Agonist-induced glycogenolysis in rabbit retinal slices and cultures. AB - 1. The effects of different putative retinal transmitters and/or modulators on glycogenolysis in rabbit retinal slices and in retinal Muller cell cultures were examined. 2. Incubation of rabbit retinal slices or primary retinal cultures (either 3-5 day-old or 25-30 day-old) in a buffer solution containing [3H] glucose resulted in the accumulation of newly synthesized [3H]-glycogen. 3. Noradrenaline (NA), isoprenaline, vasoactive intestinal peptide (VIP), 5 hydroxytryptamine (5-HT) and 8-hydroxy-dipropylaminetetralin (8-OH-DPAT) stimulated the hydrolysis of this newly formed 3H-polymer. The potency order of maximal stimulations was: VIP greater than NA greater than isoprenaline greater than 5-HT greater than 8-OH-DPAT. 4. The putative retinal transmitters, dopamine, gamma-aminobutyric acid (GABA), glycine and taurine and the muscarinic agonist carbachol (CCh) had no effect on [3H]-glycogen content. 5. The glycogenolytic effects of NA/isoprenaline and 5-HT/8-OH-DPAT appear to be mediated by beta adrenoceptors and 5-HT1 receptors (possibly 5-HT1A), respectively while the VIP induced response involved another receptor subtype. 6. Agonists which mediated [3H]-glycogen hydrolysis also stimulated an increase in adenosine 3':5'-cyclic monophosphate (cyclic AMP) formation. Both responses are blocked to a similar extent by the same antagonists and so are probably mediated via the same receptor subtypes. Moreover, dibutyryl cyclic AMP (db cyclic AMP) promoted tritiated glycogen breakdown in the three retinal preparations. 7. Not all receptors linked to cyclic AMP production however promote glycogenolysis. Dopamine and apomorphine stimulated cyclic AMP formation via D1-receptors without influencing glycogenolysis. These receptors are exclusively associated with neurones. PMID- 2568146 TI - ICI 204448: a kappa-opioid agonist with limited access to the CNS. AB - 1. A number of compounds were evaluated in an attempt to identify a kappa-opioid receptor agonist with limited access to the central nervous system. 2. Quaternary derivatives of the kappa-opioid agonists tifluadom, U-50488H and ethylketocyclazocine were essentially devoid of opioid activity in a range of isolated tissue preparations. 3. A novel compound - ICI 204448 - is described which produced a potent and naloxone-reversible inhibition of electrically-evoked contraction of the guinea-pig ileum, mouse vas deferens and rabbit vas deferens preparations. ICI 204448 was shown to displace the binding of the kappa-opioid ligand [3H]-bremazocine from guinea-pig cerebellum membranes. 4. Ex vivo binding studies in mice showed ICI 204448 to be well absorbed following subcutaneous administration. The brain levels achieved by ICI 20448 were substantially lower than those produced by kappa-agonists such as U-50488H and tifluadom. 5. A good correlation was found for a range of opioids between lipophilicity and degree of CNS penetration. PMID- 2568148 TI - Sulphasalazine in rheumatoid arthritis: haematological problems. PMID- 2568149 TI - Profound leucopenia with Salazopyrin EN. PMID- 2568151 TI - Difference in recovery patterns of striatal dopamine content, tyrosine hydroxylase activity and total biopterin content after 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) administration: a comparison of young and older mice. AB - Striatal dopamine (DA) content, tyrosine hydroxylase (TH) activity, and total biopterin content were measured as parameters of recovery, after administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in 7-week-old (young) and 28-week-old (older) C57 BL/6 mice. After 10 consecutive days of injection of MPTP (30 mg/kg/day) young mice were sacrificed at one day and 2, 4, 8, 12 and 20 weeks; older mice (20 mg/kg/day) at one day and 4, 8, and 12 weeks. All 3 parameters were markedly reduced one day after the last injection of MPTP. During the observation period, the parameters showed a gradual and partial recovery. The recovery rates of the 3 parameters differed significantly, especially during the early phases of 2-8 weeks. Total biopterin content showed a greater rate of recovery than TH activity and TH activity, a greater rate of recovery than DA content. In the older mice group, the recovery of all 3 parameters was retarded significantly, and dissociation of the recovery rates between the 3 parameters was more prominent. The results of our present study suggest that, following neurotoxic injury, the recovery of biopterin may play a significant role in dopaminergic terminal regeneration. PMID- 2568150 TI - Modulation of central nervous system actions of corticotropin-releasing factor by dynorphin-related peptides. AB - Corticotropin-releasing factor (CRF) and dynorphin-related peptides are co localized within a subset of hypothalamic neurons suggesting the possibility of their co-release. Therefore, studies were performed in conscious unrestrained rats to examine whether dynorphin-related peptides modify the central nervous system (CNS) actions of CRF on sympathetic nervous activity and cardiovascular function. Intracerebroventricular (i.c.v.) administration of dynorphin A1-8 (0.1 and 1.0 nmol) did not alter arterial pressure (AP) or heart rate (HR). I.c.v. injection of dynorphin A1-13 (0.1, 0.3 and 1.0 nmol) produced transient elevations of HR but did not significantly affect AP. I.c.v. administration of dynorphin A1-17 (0.1, 0.3 and 1.0 nmol) elicited delayed (10-15 min) and transient elevations of AP and HR. CRF (0.15 nmol, i.c.v.) produced immediate and sustained elevations of AP, HR and plasma catecholamine levels. Upon simultaneous administration, 0.1 nmol of dynorphin A1-17, but not 0.1 nmol of dynorphin A1-8 or dynorphin A1-13, markedly attenuated CRF-induced elevations of AP, HR, and plasma catecholamine levels. The results suggest that selected dynorphin-related peptides may modify the CNS actions of CRF. PMID- 2568152 TI - CNQX (6-cyano-7-nitroquinoxaline-2,3-dione) antagonizes NMDA-evoked [3H]GABA release from cultured cortical neurons via an inhibitory action at the strychnine insensitive glycine site. AB - The novel glutamate antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) inhibited glutamate stimulated [3H]GABA release from cortical neurons in vitro. Kainate-induced release was blocked in a competitive fashion but N-methyl-D aspartate (NMDA)-induced release was blocked non-competitively by CNQX. 7 Chlorokynurenate (7-CK) also inhibited NMDA evoked [3H]GABA release non competitively, but had no effect on kainate induced release. The effects of both CNQX and 7-CK on NMDA-induced release were reversed by addition of exogenous glycine but the effects of CNQX on kainate-induced release were not altered by glycine. This suggests that both CNQX and 7-CK may interact with the glycine regulatory site of the NMDA receptor. PMID- 2568153 TI - Stereotypies elicited by injection of N-propylnorapomorphine into striatal subregions and nucleus accumbens. AB - Injection of the dopamine (DA) agonist R-(-)-N-n-propylnorapomorphine (NPA; 5-40 micrograms) into anterior ventral striatal sites (either lateral (VL) or medial (VM) elicited dose-dependent oral and sniffing stereotypies of rapid onset, long duration and high intensity. In contrast, injection into anterior dorsolateral (DL) or posterior ventral (lateral (PL) or medial (PM] sites produced little oral and moderate sniffing behavior of slower onset, shorter duration and low intensity. Injection into the dorsomedial (DM) striatum produced intermediate effects. Intra-accumbens NPA elicited weak oral activity and moderate sniffing which was similar in onset, duration and intensity to the least sensitive striatal sites (DL, PM and PL). In other experiments, DA receptors were inactivated with the irreversible blocking agent N-ethoxycarbonyl-2-ethoxy-1,2 dihydroquinoline (EEDQ; 6 mg/kg) and behavioral recovery was monitored by challenge with 20 micrograms NPA into the VL or the nucleus accumbens (NA) at various times after EEDQ. Sniffing behavior recovered rapidly (normal by day 4 in both regions), whereas oral activity required 8 (NA) and 12 days (VL) to return to control levels. The results are discussed in terms of a possible topographic distribution of behavior in the striatum. Alternatively, heterogeneity of DA receptor density may account for these findings. PMID- 2568155 TI - Coexistence of enkephalin and somatostatin in the chicken retina. AB - A double immunostaining method was used in the present study to demonstrate that enkephalin (ENK) and somatostatin (SOM) coexist within single amacrine cells of the chicken retina. The ratio of double-labeled cells to either ENK- or SOM bearing cells was more than 90%, indicating that retinal ENK and SOM are most likely to be found together in the same amacrine cell. This rate of colocalization is the highest yet observed among any neurotransmitters and modulators that coexist. PMID- 2568154 TI - Ventral pallidal neuronal responses to dopamine receptor stimulation in the nucleus accumbens. AB - The possible role of ventral pallidum (VP) in expressing dopaminergic actions in the nucleus accumbens was studied electrophysiologically using extracellular single unit recording and iontophoretic techniques in urethane-anaesthetized rats. Microinjections of dopamine (130 mM, 5-10 micrograms/0.2-0.4 microliters) into the nucleus accumbens resulted in a gradual, but prolonged, increase in the firing rate of VP neurones. Injections of the D1 agonist SKF38393 (34 mM, 2 micrograms/0.2 microliters), followed by the D2 agonist quinpirole (40 mM, 2 micrograms/0.2 microliters) into the accumbens, but not in the reverse order, resulted in a similar increase in the activity of VP neurones, mimicking the dopaminergic effect. Injections of either the D1 or the D2 agonist alone into the accumbens, however, produced no significant changes. Furthermore, iontophoretic application of picrotoxin, a gamma-aminobutyric acid (GABA) antagonist, or naloxone, an opiate (including enkephalin) antagonist on the same VP neurone which responded to accumbens dopamine injection also increase its spontaneous firing rate. Thus, pre-activation of D1 receptors in the accumbens was essential for the subsequent physiological expression of D2 receptors in inducing an increase in the firing rate of VP neurones. Dopamine in the accumbens may suppress the tonic inhibitory GABAergic and enkephalinergic outputs to the VP, resulting in an increase in firing rate of VP neurones. Since previous behavioural studies have shown that dopaminergic stimulation in the accumbens increases locomotor activity, the increased firing rate of ventral pallidal neurones may be expressing the postsynaptic actions of dopamine receptor stimulations in the accumbens as initiation of locomotor activity. PMID- 2568156 TI - Regional, reversible ultrastructural changes in rat brain with chronic neuroleptic treatment. AB - Administration of the dopamine receptor antagonist (neuroleptic, antipsychotic), haloperidol, resulting in an increase in the number of dopamine binding sites in the striatum and nucleus accumbens, has been well established. These increases disappear following withdrawal of treatment. Ultrastructurally, we found an increase in the number of synapses containing perforated postsynaptic densities (PSDs) following haloperidol administration within the caudate nucleus but not within the nucleus accumbens. The effect in the caudate reversed following cessation of treatment. We speculate that the terminals undergoing the change are not dopaminergic but may originate from the cerebral cortex. This reversible morphological increase associated with dopamine antagonist drug therapy may be reflective of the tolerance developed to neuroleptic drug-induced extrapyramidal syndromes and/or may be associated with abnormal motor movements of tardive dyskinesia that occur following long-term treatment. PMID- 2568157 TI - Somatostatin increases intracellular Ca2+ concentration in cultured rat hippocampal neurons. AB - Changes of intracellular Ca2+ concentration [( Ca2+]i) in response to somatostatin were measured in cultured rat hippocampal neurons on a single cell basis by fura-2 fluorometry. Somatostatin increased [Ca2+]i dose-dependently and this effect was completely blocked in either Ca2+-depleted medium or LaCl3 containing medium. In addition, omega-conotoxin GVIA completely inhibited the effect of somatostatin. Our results indicate that somatostatin receptors couple with N-type voltage-sensitive Ca2+ channels in cultured rat hippocampal neurons. PMID- 2568159 TI - Long-term effects of cholinergic basal forebrain lesions on neuropeptide Y and somatostatin immunoreactivity in rat neocortex. AB - The effect of cholinergic basal forebrain lesions on immunoreactivity to somatostatin (SOM-i) and neuropeptide-Y (NPY-i) was investigated in the rat parietal cortex, 16-18 months after multiple bilateral ibotenic acid injections in the nucleus basalis complex. As a result of the lesion, the cholinergic fiber density in the cortex decreased by 66% with a concurrent increase in SOM-i fibers by more than 50% and a 124% increase in NPY-i fiber innervation. The neuropeptidergic sprouting response on cholinergic denervation does not match the concurrent cholinergic and peptidergic decline in Alzheimer's disease and as such does not support the cholinergic lesion alone as an animal model for this neurodegenerative disorder. PMID- 2568158 TI - Quinoxaline derivatives are high-affinity antagonists of the NMDA receptor associated glycine sites. AB - Membranes from rat telencephalon contain strychnine-insensitive glycine binding sites associated with NMDA receptors. Three quinoxaline derivatives, among them the high-affinity AMPA receptor antagonists CNQX and DNQX, were found to inhibit [3H]glycine binding to these sites with micromolar affinities. Binding of these compounds to the glycine site also inhibited glutamate-stimulated association and dissociation of [3H]TCP. This suggests that these AMPA antagonists, like the structurally related compound kynurenate, act as glycine site antagonists. PMID- 2568160 TI - Munchausen syndrome/bulimia by proxy: ipecac as a toxin in child abuse. PMID- 2568161 TI - Metabolic and endocrine responses to prolonged exercise in rats under beta 2 adrenergic blockade. AB - The respective roles of allosteric regulators and catecholamines in the control of muscle glycogen breakdown during exercise remain a matter of controversy. This study was designed to reassess the role of the sympathoadrenal system during prolonged exercise in rats. Animals were studied at rest or after treadmill exercise (28 m.min-1; 8% slope) to exhaustion in a control situation or following administration of a specific beta 2-adrenergic receptor antagonist (ICI 118,551, 1 mg.kg-1, i.v.). Running times to exhaustion were 54 and 36 min in control and treated rats, respectively. For the purpose of comparison, another group of control rats was studied after a 36-min exercise bout. The reduction in endurance in treated rats was associated with an impairment in glycogen utilization, as measured by muscle glycogen stores, in soleus muscle but not in superficial vastus lateralis or gastrocnemius lateralis muscles. Utilization of liver glycogen stores was similar in the two groups of animals, but plasma glucose (7 vs. 13 mM) and lactate (4 vs. 7 mM) levels were significantly lower in rats under beta-blockade than in control rats run for 36 min. Plasma free fatty acid and glycerol concentrations were not significantly different between groups. On the other hand, plasma epinephrine concentration was significantly higher in treated rats (13 vs. 5 mM), which might reflect a compensatory increase in adrenal activity. These results suggest that glycogen breakdown during prolonged exercise is under the control of the sympathoadrenal system in predominantly slow-twitch but not in predominantly fast-twitch muscles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568162 TI - Characterization of pyrimidine metabolism in the cellular slime mold, Dictyostelium discoideum. AB - The arginine-independent, de novo biosynthetic pathway of pyrimidines in Dictyostelium discoideum is initiated by a class II carbamoyl-phosphate synthetase (EC 6.3.5.5) specific for pyrimidine biosynthesis which utilized L glutamine as its N donor and was partially inhibited by both UTP and CTP. The second step in the de novo pathway was provided by an unregulated aspartate transcarbamoylase (EC 2.1.3.2) which primarily appeared as a multimeric enzyme of 105 kilodaltons. The next enzyme, dihydroorotase (EC 3.5.2.3), was approximately 90-100 kilodaltons. Although the early enzymatic activities of the pyrimidine pathway appeared to reside in independent protein complexes, various unstable molecular species were observed. These structural variants may represent proteolytic fragments of a multienzyme complex. In addition to de novo synthesis, the amoeba demonstrated the capacity for salvage utilization of uracil, uridine, and cytidine. Upon starvation on a solid substratum, axenically grown amoebas began a concerted developmental program accompanied by a restructuring of nucleotide metabolism. The absolute levels of the ribonucleotide pools droppedby 98% within 30 h; however, both the adenylate energy charge and the GTP/ATP ratios were maintained for 50 h after the initiation of development. The maintenance of these metabolic energy parameters required the tight cell-cell contact necessary for development, and the capacity for pyrimidine metabolism was maintained throughout developmental morphogenesis. PMID- 2568164 TI - Long-term endocrine toxicity of myeloablative treatment followed by autologous bone marrow/blood derived stem cell transplantation in patients with malignant lymphohematopoietic disorders. AB - The effect of myeloablative treatment with autologous bone marrow transplantation (ABMT)/autologous blood derived stem cell transplantation (ABSCT) in patients with acute leukemias or lymphomas was studied in 32 adult patients with a mean observation time of 15.8 months after transplantation. The conditioning regimen consisted of hyperfractionated total-body irradiation (TBI) and high-dose cyclophosphamide or the cyclophosphamide, carmustine, and etoposid (CBV) regimen. In all of the female patients, we observed primary ovarian failure requiring estradiol replacement therapy. In all of the male patients, testosterone levels were normal but follicle stimulating hormone (FSH) levels were increased, suggestive of germinal aplasia which was proved by semen analysis in several patients. In contrast to the reports of other groups, we did not find any abnormalities in thyroid function, most likely because TBI was hyperfractionated. Moderate toxicity to the adrenal cortex was noticed and was more pronounced in women than in men. Our results are similar to findings reported after allogeneic bone marrow transplantation, with the exception of normal thyroid function in our patients. These results should be taken into consideration when counseling patients about the long-term consequences of myeloablative treatment. Cryopreservation of semen should be offered to men before myeloablative treatment. Estrogen replacement should be initiated after transplantation in women to prevent adverse effects of long-term ovarian failure. PMID- 2568165 TI - Multiple endocrine syndrome type I. Clinical, laboratory findings, and management in five families. AB - The clinical features of 20 patients from five families with multiple endocrine neoplasia syndrome type I (MEN-I) were studied. Nineteen patients (95%) had hyperparathyroidism. Five patients who had a diagnosis during surgery of adenoma and who had fewer than 3.5 glands removed had recurrence of hypercalcemia after surgery. Fourteen patients (70%) had pancreatic islet cell tumors. All had one or more elevated serum polypeptide hormones, and six had symptoms related to the hormones produced. Multiple pancreatic tumors were identified in the nine patients who underwent surgery. Three patients who died had a mean survival of 6.3 +/- 2.9 years. Eight patients had pituitary tumors; seven had macroadenomas. Of the eight patients with pituitary tumors, seven had high serum prolactin and responded to bromocriptine therapy, whereas the eighth patient had acromegaly treated with radiotherapy. It was concluded that hypercalcemia due to hyperparathyroidism in MEN-I syndrome patients should be managed by a resection of four glands and transplantation of one half gland into the forearm because none of the patients has shown evidence of a recurrence, and serum calcium levels have been normal. Pancreatic tumors, which are usually multiple, may be asymptomatic. Patients with these tumors usually have long survival rates, even with distant metastasis. Total pancreatectomy may be the method of choice, especially in patients with gastrinoma caused by the diffuse nature of the disease. Long-term follow-up is needed, however, with more patients. Pituitary tumors are primarily prolactin-producing tumors, and medical treatment is the method of choice. PMID- 2568166 TI - Low incidence of loss of chromosome 10 in sporadic and hereditary human medullary thyroid carcinoma. AB - Genetic linkage has been recently documented between a centromeric region of chromosome 10 and familial multiple endocrine neoplasia type II (MEN II). This syndrome consists of initial thyroid C-cell and adrenal chromaffin cell hyperplasia which result in multifocal medullary thyroid carcinomas and bilateral adrenal pheochromocytomas. Other hereditary cancers, such as retinoblastoma, appear to result from a series of genetic events involving, first the inheritance of a germ line abnormality, and subsequent loss of chromosome loci opposite this initial defect. In these cancers, this loss of the normal alleles in both familial and sporadic cases, is frequently manifest as a reduction to homozygosity for polymorphic DNA markers near the involved locus. It might then be expected that chromosome 10 regions would be lost with high frequency in tumor DNA from patients with MEN II and sporadic medullary thyroid carcinoma (MTC). We now demonstrate that only two of 16 MTC tumors studied by analysis of restriction fragment length polymorphisms for multiple regions of the short and long arms of chromosome 10 showed loci reduced to homozygosity. One of these tumors was from a patient with MEN II and the other from a patient with nonfamilial MTC. Importantly, no such chromosome 10 changes were noted in pheochromocytomas from the patient with MEN II or his sister. These findings strongly suggest that the sequence of genetic events for familial MTC is either different from that for retinoblastoma or that loss of normal alleles opposite the germ line genetic defect occurs by mechanisms other than gross loss of chromosomal material in MTC. A model is proposed suggesting that the mechanism involving loss of alleles opposite one another is operative in hereditary tumors, such as retinoblastoma, which do not arise within a setting of initial polyclonal cellular hyperplasia. In contrast, in tumors such as familial MTC and polyposis coli which arise as individual clones of neoplastic cells from a setting of preexistent polyclonal hyperplasia, the first genetic event may underlie hyperplasia, and additional events, frequently at other chromosomal loci, may cause individual clonal neoplasms. PMID- 2568167 TI - Role of glutathione and dependent enzymes in anthracycline-resistant HL60/AR cells. AB - We studied the cellular enzymatic defenses against anthracycline-induced free radical damage in the HL60 human myelogenous leukemia cell line and in its anthracycline-resistant subline, HL60/AR. Intracellular glutathione (GSH) levels and gamma-glutamyl transpeptidase activity were lower in HL60/AR than in HL60 cells. Glutathione-S-transferase (GST) and glutathione peroxidase activities were similar in both cell lines. The intracellular distribution of GSH/GST was visualized by digitized video fluorescence microscopy, utilizing the fluorescent probe monochlorobimane fluorescence microscopy, utilizing the fluorescent probe monochlorobimane (MBCl), which is specifically conjugated to GSH by GST. In HL60 cells stained with the MBCl probe, a bright diffuse cytoplasmic and nuclear fluorescence pattern was observed, whereas in HL60/AR cells, the fluorescence was mostly localized to the Golgi apparatus with a lesser component of diffuse cytoplasmic and nuclear fluorescence. Pretreatment of HL60/AR cells with buthionine sulfoximine (BSO) partially reversed resistance to daunorubicin. This effect of BSO on resistance was associated not only with the abolition of localized MBCl fluorescence to the Golgi apparatus but also with increased intracellular accumulation and retention of daunorubicin. The results of our studies demonstrate that inhibition of GSH synthesis in HL60/AR cells results in significant sensitization to daunorubicin and suggest that changes in the intracellular distribution of GSH/GST and/or increased drug retention may be involved in mediating this effect. PMID- 2568168 TI - Heterogeneous expression of erbB-2 messenger RNA in human breast cancer. AB - Amplification and mRNA expression of the erbB-2 gene was analyzed in 61 samples of primary human breast carcinoma. In the 57 samples where RNA could be isolated four different expression level groups were identified. Comparison of hybridization signal with that for beta-actin revealed that erbB-2 mRNA could not be detected in 6 of 57 samples (11%), was detected at normal levels in 32 of 57 samples (56%), showed 4- to 8-fold overexpression in 8 of 57 samples (14%), and showed 16- to 128-fold overexpression in 11 of 57 samples (19%). Examination of the DNA of the same set of samples revealed 6 of 61 samples (10%) with distinct gene amplification and 6 of 61 samples (10%) with possible gene amplification. The highest levels of erbB-2 overexpression were associated with gene amplification. Samples with 4- to 16-fold overexpression of the erbB-2 mRNA occurred without evident gene abnormalities. There was no association of erbB-2 expression or gene amplification with clinical stage of breast carcinoma or axillary lymph node involvement. The clear amplification of the erbB-2 gene may be associated with a significantly shorter time to treatment failure. PMID- 2568169 TI - Loss of constitutional heterozygosity in colorectal tumors from patients with familial polyposis coli and those with nonpolyposis colorectal carcinoma. AB - Familial polyposis coli (FPC) is an autosomal dominant tumorigenic disorder, the major gene of which is mapped to chromosome 5q. We searched for a gene loss in colorectal tumors from FPC patients, as related to tumorigenesis by inactivation of tumor suppression genes, using restriction fragment length polymorphism analysis. The findings were compared with those in the case of nonpolyposis colorectal carcinomas (NPCC). We examined specimens from 39 FPC patients, including 21 adenocarcinomas and 49 adenomas, and 23 colorectal carcinomas from 22 NPCC patients. For this, we used 53 polymorphic DNA markers on all autosomes. Frequent loss of heterozygosity in colorectal carcinoma from FPC patients was observed on chromosomes 5 (24%), 14 (20%), 17 (31%), 18 (40%), and 22 (35%) and also on chromosomes 5 (32%), 14 (30%), 17 (27%), 18 (20%), and 22 (19%) in NPCC. Although loss of heterozygosity in adenoma from FPC patients was observed on nine chromosomes, the frequencies were less than 7%. As we fractionated tumors only macroscopically, actual frequencies of loss of heterozygosity are probably somewhat higher. However, these results do suggest that tumor suppression genes for colorectal carcinoma may locate on chromosomes 5, 14, 17, 18, and 22 and that they may play a critical role in carcinogenesis in both FPC and NPCC patients. PMID- 2568170 TI - Modification of myc gene amplification in human somatic cell hybrids. AB - In order to study whether cell fusion would modify the DNA copy number of an amplified oncogene, somatic cell hybrids were made between the human neuroepithelioma cell line MCIXC and HeLaCOT human adenocarcinoma cells. MCIXC contains approximately 21 copies of the c-myc oncogene and HeLaCOT contains approximately 5 copies relative to the control. All hybrid clones investigated displayed a marked decrease in the number of copies of c-myc DNA (an average of 5 copies), while the level of c-myc RNA in the hybrids was similar to that found in both parents. All eight hybrid clones were found to be completely nontumorigenic even though both parent cells formed tumors in 100% of the nude mice treated by injection. This loss of oncogene amplification in the hybrids was shown not to be due to either heterogeneity of c-myc amplification in the MCIXC parent or segregation of a copy of the chromosome 22 from the hybrids. This loss most likely resulted from the breakdown of a homogeneously staining region (containing the amplified gene copies) into double minutes, which were subsequently lost from the cells. The HeLaCOT cell line was also fused to the human neuroblastoma BE(2)C, which contains approximately 123 copies of the N-myc oncogene relative to control. Ten hybrid clones were found to contain an average of 47 copies of N-myc DNA, significantly less than the 91 copies predicted had no loss occurred. These BE(2)C x HeLaCOT hybrids expressed on average about 15% the N-myc RNA seen in the BE(2)C parent and, as with the MCIXC x HeLaCOT hybrids, were found to be completely nontumorigenic. However, upon passage in culture, one BE(2)C x HeLaCOT hybrid eventually became tumorigenic. This hybrid also displayed reduced copies of N-myc DNA in comparison to its parent hybrid but surprisingly showed a 2-fold increase in N-myc RNA. Thus, the expression of N-myc, but not the amplification state of either myc gene, appears to correlate with the tumorigenicity of the cells. PMID- 2568163 TI - Inflammatory bowel disease. AB - An increasing number of options are available for the treatment of inflammatory bowel disease; the selection depends on the extent and severity of the disease. Experience with sulfasalazine and corticosteroids has led to a proliferation of 5 aminosalicylic acid (5-ASA) compounds and experimentation with alternative corticosteroid preparations. Given rectally 5-ASA is particularly effective in the treatment of distal ulcerative colitis, and experience is accumulating with several oral formulations. Metronidazole is useful in some cases, and immunosuppressive agents have a role in some patients with chronic refractory disease. A variety of measures, such as nutritional therapy, surgery and psychosocial support, are important elements of therapy. Further therapeutic innovations are expected as the etiology and pathogenesis are clarified. PMID- 2568171 TI - Characterization of a K562 multidrug-resistant cell line. AB - A daunorubicin-resistant variant of the K562 human leukemia cell line (K562-R), which demonstrates cross-resistance to other anthracycline antibiotics and Vinca alkaloids, has been developed in vitro by continuous exposure to daunorubicin. Cross-resistance to anthracyclines and Vinca alkaloids is reversed when cells are exposed to drugs in the presence of verapamil, a calcium channel blocker. The K562-R cell line overexpresses a 4.5-kilobase mRNA, which is thought to code for the Mr 170,000 membrane glycoprotein associated with multidrug resistance. Transport studies indicate reduced intracellular accumulation and retention of daunorubicin in the K562-R cells as compared to the parent cell line. These studies further suggest the presence of distinct cellular pools composed of both rapidly and slowly exchanging drug, with the rapidly exchanging pool being more pronounced in the resistant line. The development of multidrug resistance in the K562-R cell line is also associated with the overexpression of five different cell surface membrane proteins ranging in molecular weight between 50,000 and 210,000, whose function remains to be defined. PMID- 2568172 TI - Multidrug resistance in mitoxantrone-selected HL-60 leukemia cells in the absence of P-glycoprotein overexpression. AB - A multidrug-resistant variant of the human HL-60 promyelocytic leukemia cell line (HL-60/MX2) has been isolated in vitro by subculturing these cells in progressively increasing concentrations of mitoxantrone. The MX2 cells are cross resistant to etoposide, teniposide, bisantrene, dactinomycin, 4'-(9 acridinylamino)methanesulfon-m-anisidide, and the anthracyclines daunorubicin and doxorubicin but retain sensitivity to the Vinca alkaloids melphalan and mitomycin C. In addition, the MX2 cells display slight collateral sensitivity to bleomycin. Despite being 30-35-fold less sensitive to mitoxantrone, net [14C]mitoxantrone accumulation at 60 min was reduced by only 10% in the mitoxantrone-resistant cells compared to the parental line. Furthermore, at later time points, e.g., 120 and 180 min, mitoxantrone accumulation in the MX2 cells exceeded that in HL-60 cells by 8.5 and 6.4%, respectively. No significant differences were observed between the sensitive and resistant cell lines in the initial (first 60 s) accumulation of mitoxantrone, and only minor (3-6%) enhancement of mitoxantrone efflux was detected in the resistant cell type. Monoclonal antibodies to P glycoprotein had no detectable reactivity with membrane vesicles from either the sensitive or resistant cell types as determined by standard immunoblotting techniques. The mitoxantrone-resistant cells displayed a reciprocal translocation [rcpt(1;3)-(q21;p23)] not found in the sensitive parent, but there were no demonstrable double minute chromosomes or homogeneous staining regions in cells from either line. Thus, these mitoxantrone-resistant human leukemia cells display many features which are atypical for the "classic" multidrug resistance phenotype and should provide a useful model for the study of multidrug resistance which is not mediated by P-glycoprotein. PMID- 2568173 TI - Alterations in Langerhans cells and Thy-1+ dendritic epidermal cells in murine epidermis during the evolution of ultraviolet radiation-induced skin cancers. AB - To understand the role of cutaneous immune cells in host resistance to the induction and growth of skin cancer, we investigated the number and morphology of murine dendritic epidermal cells (dEC) during the evolution of ultraviolet (UVA) UV-induced skin cancers. Female C3H/HeN mice were treated topically with 8 methoxypsoralen followed by ultraviolet A (UVA) radiation 3 times/week or irradiated with UVB radiation 3 times/week. In both psoralen plus UVA- and UVB treated mice, ATPase+ and Ia+ Langerhans cells almost completely disappeared from the treated skin during the early latency period of tumor development (4 weeks) but reappeared in the epidermis late in the latency period (between 15 and 22 weeks). The ATPase+ cells that reappeared in the epidermis had a rounder, less dendritic morphology than normal Langerhans cells. Thy-1+ dEC were totally depleted from the epidermis in both treatment groups at the end of first week of treatment and were nearly absent from the skin during the entire latency period. After tumors appeared (29 weeks), Thy-1+ dEC were still absent or detected only in small numbers in skin surrounding the tumors. ATPase+ and Ia+ cells present in skin around the tumors constituted 60 to 80% of the number in nonirradiated skin. Mice that received UVA radiation alone developed no tumors. ATPase+ and Ia+ Langerhans cells and Thy-1+ dEC were detected in UVA-treated epidermis after 22 weeks and 43 weeks, although the numbers were lower than those in unirradiated mice. Most psoralen plus UVA-induced tumors (81%) were squamous cell carcinomas, whereas only 24% of UVB-induced tumors were of this histological type. Our results demonstrate that UV-induced skin cancers developed in the presence of ATPase+ and Ia+ cells in the epidermis and in the absence of Thy-1+ dEC. PMID- 2568174 TI - Importance of Lyt 2+ T-cells in the curative effectiveness of a low dose of melphalan for mice bearing a large MOPC-315 tumor. AB - We have previously demonstrated that the curative effectiveness of a low dose (2.5 mg/kg) of melphalan (L-phenylalanine mustard; L-PAM) for mice bearing a large s.c. (approximately 20 mm in diameter) MOPC-315 tumor and extensive metastases requires the participation of T-cell-dependent antitumor immunity in tumor eradication (S. Ben-Efraim et al., Cancer Immunol. Immunother., 15: 101 107, 1983). Here we show that the Lyt 2+ T-cells, and not the L3T4+ T-cells, participate in the cure of such tumor-bearing mice by a low dose of L-PAM. Specifically, depletion of Lyt 2+ T-cells from mice bearing a large MOPC-315 tumor by treatment with monoclonal anti-Lyt 2.2 antibody abolished the curative effectiveness of the low dose of drug. In contrast, depletion of L3T4+ T-cells from mice bearing a large MOPC-315 tumor by treatment with monoclonal anti-L3T4 antibody did not reduce significantly the curative effectiveness of the low dose of drug. Histological examination of tumor nodules on various days following low dose L-PAM therapy revealed widespread lymphocytic infiltration by Day 5 following the chemotherapy, and this infiltration was drastically reduced when the L-PAM-treated tumor bearers were treated with either anti-Thy 1.2 or anti-Lyt 2.2 antibody but not with anti-L3T4 antibody. The antitumor immunity exhibited by Lyt 2+ T-cells derived from mice which were in the process of eradicating a large MOPC-315 tumor following low-dose L-PAM therapy was exploited successfully to confer systemic antitumor immunity to mice bearing a barely palpable tumor. Specifically, the adoptively transferred Lyt 2+ splenic T-cells, in conjunction with a subcurative dose of L-PAM, brought about the cure of most mice. The Lyt 2+ splenic T-cells from L-PAM-treated MOPC-315 tumor bearers were also found to be capable of exerting a direct potent lytic effect against MOPC-315 tumor cells in an antigen-specific manner. Thus, it is conceivable that the direct cytotoxic activity of Lyt 2+ T-cells for MOPC-315 tumor cells is responsible, at least in part, for the ability of the Lyt 2+ T-cells from L-PAM-treated MOPC-315 tumor bearers to bring about the eradication of the tumor burden not eradicated through the direct antitumor effects of the low dose of drug. PMID- 2568176 TI - Which atypical antipsychotics are identified by screening tests? AB - Only two tests were specific for antipsychotic potential. All effective antipsychotics blocked pharmacologically induced locomotion and affected firing in the mesolimbic DA neurons. The remaining single- (Table 1) and repeated- (Table 2) dose tests identified the atypical antipsychotics. Clozapine, thioridazine, sulpiride, tiospirone, and molindone were atypical in both types of study. Pimozide, pipamperone, aceperon, methylperon, and clotiapine were atypical in single-dose studies, clopenthixol in repeated-dose studies. Since the biochemical abnormality causing psychoses is unknown, it may be that current methods of screening for new antipsychotics are inadequate and possibly inappropriate. If a neurotransmitter other than DA is the primary cause, then totally new tests may be needed. Present tests, especially those involving behavioral paradigms, may continue to select out compounds that cause EPS side effects. New methods such as positron emission tomography scanning and other brain-imaging techniques hold the promise of studying specific types and subtypes of receptors in the living human brain. The recent discovery of chromosomal abnormalities in psychotic illness may provide new insights into the biochemical causes of such disorders and lead to completely new compounds that will be both safer and more effective. In the meantime we plan to review the literature on the clinical use of the compounds identified as atypical in this article and to develop research protocols to assess their efficacy in treatment-resistant psychoses and intractable conditions such as tardive dyskinesia. PMID- 2568177 TI - Trazodone: from the mental pain to the "dys-stress" hypothesis of depression. AB - Trazodone was developed according to the mental pain hypothesis, which was postulated from studying patients and which proposes that depression is associated with a decreased pain threshold. Trazodone is devoid of the typical aminergic properties of tricyclics and monoamine oxidase inhibitors. Its preeminent effects are increased pain threshold and alpha-adrenergic blockade. The "dys-stress" hypothesis maintains the concept of the decreased pain threshold in depression, but attributes it to a pathology of the stress response. Whereas physiologically this response produces various effects, including analgesia and alertness that improve the mental and physical performance, in some individuals it is impaired. Abnormalities of the stress response are proposed to be a predisposing or pathogenetic factor for depression and other conditions. According to the "dys-stress" hypothesis, the alpha-adrenergic blockade produced by trazodone and its congeners would also be implicated in its antidepressant activity, as well as its side effects and preferential uses in depressive states associated with adrenergic hyperactivity. PMID- 2568175 TI - Phase I and pharmacodynamic study of taxol in refractory acute leukemias. AB - Taxol, a novel antimicrotubule agent that enhances tubulin polymerization and microtubule stability, was administered to adults with refractory leukemias as a 24-h i.v. infusion in a Phase I study. The primary objectives were to determine the maximum tolerated dose of taxol administered on this schedule to patients with acute leukemias and describe the nonhematological toxicities which became dose limiting. The starting dose, 200 mg/m2, was based on the maximum tolerated dose in solid tumor trials, in which myelosuppression precluded dose escalation. Seventeen patients received 28 evaluable courses at 200, 250, 315, and 390 mg/m2. Severe mucositis limited further dose escalation. Other nonhematological effects included peripheral neuropathy, alopecia, myalgias, arthralgias, nausea, vomiting, diarrhea, and an acute pulmonary reaction that was presumptively due to taxol's Cremophor vehicle. Mean peak taxol plasma concentrations at all dose levels were in the range of concentrations that were previously demonstrated to induce microtubule bundles, a morphological effect associated with cytotoxicity, in leukemia cells in vitro. Pretreatment blasts from 12 patients were incubated with taxol ex vivo. Taxol-induced microtubule bundles were apparent in the blasts of eight patients who also had cytoreduction of tumor, and sensitivity to bundle formation was related to the magnitude of antitumor activity. In contrast, taxol did not induce microtubule bundles ex vivo in the blasts of the other four total nonresponders. Based on this study, the maximum tolerated doses and recommended Phase II doses for taxol, limited by nonhematological toxicity and administered as a 24-h i.v. infusion to patients with refractory leukemias, are 390 and 315 mg/m2. Phase II trials at these myelosuppressive doses are required to determine taxol's activity in the treatment of leukemias. In addition, further evaluation of microtubule bundle formation ex vivo in Phase II studies is necessary to determine the ultimate utility of this assay in assessing tumor sensitivity to taxol. PMID- 2568178 TI - A community-wide perspective of secular trends in the therapeutic management of patients with acute myocardial infarction. The Worcester Heart Attack Study. AB - As part of an ongoing community-wide study of time trends in the incidence and case-fatality rates of patients hospitalized with acute myocardial infarction (MI) in 16 Worcester, Mass., metropolitan hospitals during the calendar years 1975, 1978, 1981, and 1984, changes over time in the therapeutic management of 3,263 patients with validated acute MI were examined. Beta-blocker and nitrate therapy use increased consistently and dramatically. Use of antiplatelet agents was inconsistent, while use of digoxin remained stable. Use of antiarrhythmic medications other than lidocaine decreased consistently while lidocaine use increased between 1975 and 1978 and then leveled off to being used in approximately 45% of hospitalized patients with acute MI in 1981 and 1984. A variety of demographic (e.g. age, sex, teaching hospital) and clinical characteristics (e.g. MI order, MI type, MI location, peak CPK findings, occurrence of acute clinical complications) were also associated with the use of these therapies. The results of this community-wide study suggest changes over time in the therapeutic management of patients hospitalized with acute MI and of various patient demographic and clinical factors associated with the use of these agents. PMID- 2568179 TI - Early intravenous beta-blockade in acute myocardial infarction. AB - The clinical effects of early intravenous beta-blockade followed by short-term oral treatment in acute myocardial infarction (MI) have been studied in 30 randomized trials totaling almost 28,000 patients. This treatment reduces the incidence of infarction by 10-15% in patients with threatened MI, reduces infarct size by 20-30%; reduces the incidence of nonfatal reinfarction, nonfatal cardiac arrest and mortality each by about 15%. Treatment has to start within 12 h of the onset of symptoms to be able to reduce measures of infarct size and infarct development. If patients are carefully selected serious side effects are rare and reversible. Based on the different presumed mechanisms of benefit it would be reasonable to expect the combination of i.v. beta-blockade and other therapies of proven benefit to be more beneficial than either class of agent used alone. PMID- 2568180 TI - Postinfarction unstable angina. Pathophysiologic basis for current treatment modalities. AB - Unstable angina is an acute coronary syndrome characterized by the rapid progression of clinical symptoms which may culminate in acute myocardial infarction, infarct extension or sudden death. The pathologic substrate involves atherosclerotic plaque rupture with platelet deposition, thrombus formation and coronary arterial spasm. Patients with postinfarction angina represent a high risk subgroup with severe multivessel disease, compromised collateral vessels and/or partially occlusive thrombi; their risk of infarct extension and death is significantly increased. Initial therapy includes nitrates, beta-adrenergic blockers, calcium channel antagonists, aspirin and possibly i.v. heparin, as well as prompt identification and control of exacerbating factors. Thrombolytic therapy may assume a more central role based on its ability to achieve rapid clinical stabilization. Percutaneous transluminal coronary angioplasty and coronary artery bypass grafting may be used emergently in patients refractory to medical therapy, or electively when clinically indicated. PMID- 2568181 TI - Morning increase of onset of myocardial infarction. Implications concerning triggering events. AB - During the past 5 years it has been clearly established that there is a prominent morning increase in the frequency of onset of acute myocardial infarction. Similar increases have also been observed for the related conditions of sudden cardiac death, stroke and episodes of transient myocardial ischemia. The period from 6 a.m. to noon is also a time when a number of physiologic processes that could contribute to the onset of coronary thrombosis are intensified. Arterial pressure, which could lead to plaque rupture, rises; coronary tone increases; and platelet aggregability, which could contribute to a hypercoagulable state, increases. The immediate significance of these observations is the emphasis that should be placed on pharmacologic protection of patients during the morning hours. The primary longer-term significance of the recognition of the morning increase of onset of acute cardiovascular disease is the contribution it makes to the concept that onset of coronary thrombosis at any time of the day is frequently triggered by the activities of the patient. Investigation of this possibility may yield more information about the mechanism of disease onset and facilitate design of more effective preventive therapy. PMID- 2568182 TI - Immunohistochemical study on fetal raphe samples transplanted into the leptomeningeal tissue of 5,6-dihydroxytryptamine-treated adult rats. AB - Pieces of fetal midbrain raphe containing serotonergic and dopaminergic neurons were transplanted into the leptomeningeal tissue of adult host rats that had previously been denervated by treatment with 5,6-dihydroxytryptamine. One, 2 and 5 months after transplantation, the rate of neuronal survival in the grafted tissue and the extent of axonal outgrowth into the host brain were studied by use of serotonin and tyrosine hydroxylase (TH) immunohistochemistry. The survival rate of the grafts in the 1-month group was approximately 70%. Neurons containing either serotonin or catecholamine were demonstrated by means of immunocytochemical procedures in the grafts. Two and 5 months after transplantation, serotonin-immunoreactive nerve fibers were densely distributed throughout the graft tissue, while TH-immunoreactive fiber elements were restricted to an area near the somata of TH-positive neurons. Numerous serotonin immunoreactive fibers derived from the transplant were found in the leptomeningeal tissue surrounding the graft, on the wall of neighboring blood vessels, and also in the adjacent parenchyma of the host brain. Outgrowing TH immunoreactive nerve fibers were not observed in the host brain, although such elements occurred in the leptomeningeal tissue and the wall of the larger blood vessels. These results suggest that the serotonergic and catecholaminergic (dopaminergic) neurons located in transplants of the raphe nuclei show different patterns when reinnervating the host tissue. PMID- 2568184 TI - Limb-threatening injuries in sport. AB - It is the closed gross deformity of the knee with vascular compromise that most commonly leads to amputation. This algorithm is offered as an approach to this problem with great medical and legal ramifications. Compartment syndrome about the elbow and foot can be avoided if the signs and symptoms are recognized and the fascial compartments released. PMID- 2568185 TI - Cold exposure injuries: prevention and treatment. AB - A variety of cold exposure injuries were discussed, including frostnip, chilblains, trench foot, frostbite, and hypothermia. The usual precipitating factors for each were discussed along with the clinical course, the pathophysiology, and management from the perspective of one being in the field or backcountry. Prevention is the key to all cold-related problems as adequate treatment is often not possible out of the hospital setting. Detailed recommendations were made for all conditions, relative to treatment and prevention, with special emphasis on hypothermia. PMID- 2568183 TI - Morphology and immunocytochemistry of two endocrine cell types in the guinea-pig esophageal epithelium. AB - Two types of endocrine cells in the basal layer of the keratinized stratified squamous epithelium in the guinea-pig esophagus were studied with immunohistochemistry by means of a streptavidin-biotin-bridge technique and by the immunofluorescence double-labelling technique. Cell-type I exhibited immunoreactivity to chromogranin A and to nucleosidetriphosphate adenosinediphosphate-(ADP)-phosphotransfera se. Ultrastructurally, this cell type was characterized by small cytoplasmic dense-core vesicles in which immunoreactive product was localized. Cell-type II contained large membrane limited granules, which were moderately electron dense. These granules displayed somatostatin immunoreactivity. Both cell types were located in close vicinity to non-myelinated nerve fibers and small blood vessels. The results provide evidence that, independent from the type of lining epithelium, the gastro-enteropancreatic system in guinea-pig extends to the lower portion of the esophagus. PMID- 2568186 TI - Trauma to the oral cavity. AB - Sports events, particularly those in which mouth protectors are not commonly used, can be associated with injuries to the oral structures. While the incidence of sports-related orofacial injuries is not as high as for other traumatic events, the incidence could be reduced further by increased use of mouth and face protection. When injuries occur, quick action by primary health care providers can significantly improve the final outcome of the definitive treatment. Also important is appropriate follow-up evaluation and care since the effect of trauma on teeth is not always immediately apparent. Destructive resorption, unnecessary tooth loss, and possible alveolar infections may be prevented by referral to appropriate health care providers for evaluation following the initial urgent care of trauma patients. PMID- 2568187 TI - A deletion of the PDC1 gene for pyruvate decarboxylase of yeast causes a different phenotype than previously isolated point mutations. AB - We deleted most of the pyruvate decarboxylase structural gene PDC1 from the genome of Saccharomyces cerevisiae. Surprisingly, mutants carrying this deletion allele showed a completely different phenotype than previously described point mutations. They were able to ferment glucose and their specific pyruvate decarboxylase activity was only reduced to 45% of the wild type level. Northern blot analysis revealed that a sequence in the yeast genome homologous to PDC1 and formerly designated as a possible pseudogene is expressed and may code for a different but closely related pyruvate decarboxylase. The products of the two PDC genes seem to form hybrid oligomers, however both homooligomers have enzyme activity. Thus, the product of the PDC1 gene is not absolutely necessary for glucose fermentation in yeast. PMID- 2568188 TI - Fetal substantia nigra grafts. Effect on dopamine receptors in the rat corpus striatium. AB - Effects of fetal substantia nigra grafts on the dopamine receptors in the corpus striatum in rats were investigated after the destruction of the nigrostriatal dopaminergic pathways with intraventricular 6-hydroxydopamine injections. The expected dopamine receptor denervation supersensitivity was demonstrated by a 53.7% increase of [3H]spiroperidol binding in rats with sham grafts compared with normal control-group rats. In contrast, rats with grafts showed a significant reduction of supersensitivity, with a 25% decrease in binding to the graft bearing caudate when compared with the sham-graft group. A nonsignificant 15% decrease in binding on the nongrafted side was also observed. The fetal substantia nigra grafts thus reduced the denervation supersensitivity toward a normal level. PMID- 2568189 TI - Treatment of chronic idiopathic urticaria with astemizole. AB - Astemizole is a new, long-acting H1 receptor antagonist that has proven effective in controlling the signs and symptoms of chronic idiopathic urticaria, without the sedative and anticholinergic side effects that typify use of many antihistamines. In an eight-week open study of 20 patients, astemizole significantly decreased the signs and symptoms of urticaria, as well as the severity of urticaria by anatomic location. Five patients reported complete clearing. Modest weight gain while on chronic therapy appears to be a significant side effect. PMID- 2568190 TI - Omentum graft for intractable subdural empyema. AB - A subdural empyema developed in a young man after craniotomy for evacuation of a hematoma in a sylvian fissure arachnoid cyst and the subdural space. Despite prolonged systemic and subdural antibiotic administration and a debridement of the subdural space, infection persisted, as evidenced by persistent fever, an elevated white blood cell count, and an extremely low cerebrospinal fluid glucose level. The infection was cured after a second debridement operation where microsurgically revascularized free omentum was used to obliterate the cyst and to cover the cerebral hemisphere in the craniotomy defect. The use of vascularized free omentum may prove useful in cases of refractory cranial wound infection and cerebrospinal fluid fistulas. PMID- 2568192 TI - Mapping the locus of autosomal dominant polycystic kidney disease: diagnostic application. AB - Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common autosomal dominant disorders affecting humans. The recent identification of a number of restriction fragment length polymorphisms linked to ADPKD now offers a method for diagnosis in families that are large enough for linkage and phase of linkage to be established. Initial studies of greater than 50 families with the disease localized all disease-producing mutations to the short arm of chromosome 16. Recently, however, families have been identified in which linkage to 16p markers cannot be detected. Such genetic heterogeneity of linkage limits the value of diagnostic methods based on linkage analysis and focuses attention on the need for direct diagnosis of disease-producing mutations. This in turn requires the isolation, cloning, and characterization of ADPKD genes. A refined map of the region around the PKD1 gene, the ADPKD gene localized on chromosome 16p, is presented. PMID- 2568191 TI - Blockade of NMDA-receptors prevents ocularity changes in kitten visual cortex after reversed monocular deprivation. AB - We investigated in the striate cortex of kittens whether the recovery from the effects of monocular deprivation that occurs after reverse occlusion requires activation of N-methyl-D-aspartate (NMDA) receptors. The right eye of 3-4-week old kittens was closed by lid suture for one week. Subsequently this eye was reopened and the left eyelid sutured closed for another week. During this second week, the NMDA-receptor antagonist, 2-amino-5-phosphonovaleric acid (APV), was infused from an osmotic minipump into the left visual cortex (50 nmol/h), while the right visual cortex was infused only with vehicle solution (saline) as control. At the end of the second week, the ocular dominance of striate cortical neurons was assessed with single unit recording. In the control hemispheres, the large majority of neurons was dominated by the newly opened eye, while in the APV treated hemispheres most neurons were still dominated by the newly deprived eye. In addition, neurons in the APV-treated hemispheres were less responsive and showed a reduction of orientation tuning. These data confirm that chronic blockade of cortical NMDA-receptors disrupts the disconnection of deprived pathways after monocular deprivation and reduces both responsiveness and orientation selectivity of cortical neurons. In addition they indicate that blockade of NMDA-receptors prevents also vision-dependent recovery of deprived pathways after reverse occlusion. PMID- 2568193 TI - Cystic fibrosis: diagnostic testing and the search for the gene. AB - Cystic fibrosis (CF) is a lethal genetic disorder inherited as an autosomal recessive at a frequency of about 1/2000 in Caucasian populations. A DNA marker genetically linked to CF was identified through a collaborative effort by random screening with a collection of RFLP markers on a set of CF families. The marker (CRI-L917) was mapped to chromosome 7. Construction of a genetic linkage map spanning the entire chromosome has led to the identification of a subset of 11 markers close to and flanking the CF locus. Using techniques of pulsed-field gel electrophoresis, which allow very large DNA fragments to be separated, we used seven probes to generate a long-range restriction map covering 12 million base pairs surrounding the CF locus. Information from the map is being used to isolate new probes closer to the CF gene. Methods being developed will allow candidate genes to be tested for their ability to correct defects in ion transport in cultured CF cells. PMID- 2568194 TI - Fundamental mechanisms of tumorigenesis in the human nervous system: isolation and characterization of genes associated with hereditary forms of cancer. AB - Many human cancers are known to occur in two different forms: as sporadic tumors in the general population and as hereditary tumors within families. Hereditary cancer syndromes offer unique model systems for isolating genes whose mutations lead to cancer. Here I briefly outline "reverse genetics" strategies for the isolation of the defective genes associated with three hereditary tumor syndromes of the human nervous system: neurofibromatosis type 1 (von Recklinghausen's disease) and type 2, and von Hippel-Lindau disease. The cloning and characterization of these genes will have important implications for diagnosis and treatment, not only for the relatively rare hereditary tumors, but also for their much more common sporadic counterparts such as the majority of sporadic brain tumors and renal-cell carcinomas. PMID- 2568195 TI - The new genetics of bipolar affective disorder: clinical implications. AB - Underrecognition and undertreatment of affective disorders (i.e., major depressive disorder and bipolar affective disorder) constitute a serious public health problem in this country. The recent availability of sufficient mapped restriction fragment length polymorphism (RFLP) probes to cover the human genome and of improved methods of genetic linkage analysis make the definition of the genetic basis of bipolar affective disorders (and recurrent depressions) feasible within the foreseeable future. However, the degree of genetic heterogeneity involved might make the use of tightly linked RFLPs for diagnostic testing, as has been done for Huntington's disease or cystic fibrosis, impractical. Assuming multiple disease loci, then widely applicable and useful "molecular" diagnostic tests will await the cloning of the disease genes or identification of pathogenic gene products. In addition, the combination of genetic heterogeneity and a substantial degree of assortative mating (i.e., people with affective disorders marrying each other) in families with these disorders could make the search for linked loci a "bottleneck" in the path toward developing such diagnostic tests. Recently, three disease loci have been tentatively identified on chromosome 6, 11, and X. PMID- 2568196 TI - Construction of a linkage map of the human genome, and its application to mapping genetic diseases. AB - A powerful method for localizing the genes responsible for genetic diseases is to "mark" each chromosomal region so as to permit its inheritance to be tracked in families carrying the disease; the disease gene must lie in the same region as any marker with which it is co-inherited. Chromosomes can be efficiently marked in this way by using restriction fragment length polymorphisms (RFLPs). We describe the construction of a map of the human genome with over 550 RFLP markers, such that over 95% of the genome is detectably linked to one or more of these markers. This map will make possible efficient searches for genes underlying a variety of disorders, both simple and complex, and lead to the development of presymptomatic diagnostic tests for these diseases. PMID- 2568197 TI - Clinical significance of genetic rearrangements in human neuroblastomas. AB - Human neuroblastomas are characterized cytogenetically by manifestations of gene amplification and by partial monosomy for the short arm of chromosome 1. Analysis of 646 neuroblastomas and 16 ganglioneuromas showed N-myc gene amplification in only seven of 169 (4%) with low stages of disease but in 138 of 436 (32%) with advanced stages of disease. N-myc amplification was seen in four of 41 (10%) stage IV-S tumors, and none of 16 ganglioneuromas. Analysis of patient survival by stage, age, and N-myc copy number indicate that N-myc amplification was highly correlated with rapid tumor progression, even in patients with low stages of disease. Partial monosomy of chromosome 1p can be more effectively detected by analysis of restriction fragment length polymorphisms to detect somatic loss of heterozygosity (LOH) for chromosome 1p, which is the molecular equivalent of chromosome deletion. Analysis of pairs of normal and tumor DNAs from patients with neuroblastoma demonstrated that 13 of 47 tumors (28%) had LOH at one or more loci on distal chromosome 1p; the region that shows LOH most consistently is between 1p36.1 and 1p36.3. LOH for 1p shows a highly significant correlation with N-myc amplification, suggesting that these two genetic events are related and characterize a genetically distinct subset of aggressive neuroblastomas. PMID- 2568198 TI - Dopamine, the kidney and essential hypertension. AB - The evidence for dopamine as an intrarenal natriuretic hormone is reviewed. Some patients with essential hypertension may have an uncoupling of the sodium to dopamine relationship in the kidney and their blood pressures may respond particularly well to dopaminergic agonists. PMID- 2568199 TI - Hyperkinetic hemodynamic pattern in some borderline hypertension: differential responses to alpha and beta agonists. AB - Twenty eight male borderline hypertensive patients(BH), mean age of 19.72 +/- 2.80 years, were studied and matched with 28 normal controls (NC). Alpha adrenergic responsiveness was estimated with DPD15 (the dosage of neosynephrine required for an increase of diastolic pressure of 15mmHg); beta-adrenergic responsiveness with CD25 (the dosage of isoproterenol required for an increase of heart rate of 25 beats/minute). Using CD25 of 1.0ug (mean of the NC), the BH could be subdivided into two specific types: 13 patients (46%) whose CD25 less than 1.0 ug, 0.50 +/- 0.24 (m +/- SD) had a hyperkinetic pattern; the remaining 15 patients (54%) whose CD25 greater than 1.0ug, 1.76 +/- 0.32 (m +/- SD), had a normokinetic one. Values of DPD15 of the two types were comparable. Cardiac index (CI, L/min/M2) and total peripheral resistance (TPR, dynes/sec/cm-5) of the two types manifested significant differences, both p less than 0.001. When plotting CI and TPR versus CD25, good correlations were obtained. In the BH group, the coefficient r was -0.73 between CI and CD25, + 0.70 between TPR and CD25; correlations were poor when plotting versus DPD15. Similar correlations also happened in the NC. The present study suggests that coexistence of two different hemodynamic patterns may be due to the imbalance resulted from the interaction between alpha-adrenergic responsiveness and beta-adrenergic responsiveness. PMID- 2568200 TI - Pressor responsiveness in essential hypertension and the effects of treatment with an alpha blocker, calcium antagonist or ACE inhibitor. AB - We have studied the contribution of neurohumoral and structural factors to pressor responsiveness and peripheral resistance in mild/moderate hypertension. Pressor responses to intravenous infusions of phenylephrine (an alpha1 agonist) and angiotensin II were studied in groups of patients with essential hypertension before and after treatment, for 6 weeks with either nifedipine (20 mg bid), enalapril (20 mg daily) or doxazosin (2 mg daily). All drugs lowered blood pressure to a similar extent. Pressor responsiveness to both phenylephrine and angiotensin II showed wide intersubject variation when expressed as the dose of agonist required to raise mean arterial pressure by 20 mmHg (PD20). A group of age-matched normotensive controls showed a similar PD20 for phenylephrine to hypertensives. Angiotensin 11 sensitivity was greater in hypertensives. Drug treatment had different effects in hypertensive patients. Doxazosin, an alpha blocker, reduced the responsiveness to phenylephrine but had no effect on responses to angiotensin II. Nifedipine attenuated responses to both agonists while treatment with enalapril increased responsiveness to both phenylephrine and angiotensin II. We have not found evidence of systematic differences in alpha 1 receptor responses in hypertensives and different "vasodilator" drugs can lower blood pressure with widely different effects on adrenergic and non-adrenergic vascular responses. PMID- 2568201 TI - Functional significance of B1- versus B2-receptors for cardiac inotropic and chronotropic responses in man. AB - Assessment of the functional significance of B2-receptors in the human heart is possible by employing combinations of B-receptor agonists and antagonists or by endogenous cardiac B-receptor stimulation combined with different B-receptor antagonists. A number of studies have shown that positive chronotropic responses indeed can be mediated by B2-receptors both in vitro and in intact humans. Regarding inotropic responses, tissue bath data in isolated human myocardium indicates that B2-receptors are coupled to a positive inotropic response. Our studies in healthy humans using the P/V ratio as an index of myocardial contractility show that B2-receptor stimulation can cause an inotropic response but less than the chronotropic response. Results obtained with B-receptor stimulation by increased cardiac sympathetic nerve activity versus B-receptor stimulation by exogenously administered agonists support the concept that the cardiac B1-receptor subpopulation may represent the "innervated" B-receptor linked to the synaptic cleft whereas the cardiac B2-receptor subpopulation may represent the "humoral" B-receptor associated with part of the cell in close contact with the circulation. PMID- 2568202 TI - Comparative effects of labetalol and bromazepam on ambulatory blood pressure of Nigerians with labile and stress hypertension. AB - Twenty Nigerians with labile essential hypertension (LEH) were asked to record their blood pressure and pulse rate for 14-16 hours using the Remler Portable Ambulatory Blood Pressure Recorder while exposing themselves to the stress of Lagos traffic. The average blood pressure (systolic and diastolic) and pulse rate for the test day (ASBP, ADBP and APR) were determined in a cross-over randomised open design before (C), after one week on placebo bidaily (P1), after one week on bromazepam 1.5 mg bidaily (B), after one week on placebo bidaily (P2) and after one week on labetalol 100 mg bidaily (L). Allocation to the active drugs was randomised. All drugs were administered at 8.00a.m. and 8.00p.m. respectively. The maximum blood pressure (systolic and diastolic) and pulse rate MxSBP, MxDBP and MxPR were similarly determined. Both B and L significantly reduced ASBP, ADBP, APR, MxSBP, MxDBP and MxPR but L produced a much greater reduction in the above parameters than B. Side effects observed included drowsiness with B (two subjects) and postural dizziness with L (one subject). Both drugs were effective in controlling LEH but L was more effective than B in reducing stress hypertension. PMID- 2568203 TI - An in vitro study of interactions between doxazosin and enalaprilat at vascular alpha 1-adrenoceptors. AB - 1. Isolated perfused male Sprague-Dawley rat tail artery segments were used to investigate interactions between the alpha-1-adrenoceptor antagonist, doxazosin, and the angiotensin converting enzyme (ACE) inhibitor, enalaprilat, using phenylephrine (PE) as the alpha 1-adrenoceptor agonist. 2. In concentrations of up to 10(-5) mol/L, enalaprilat had no effect on arterial responses to PE. 3. Doxazosin produced a concentration-dependent competitive alpha 1-adrenoceptor antagonism, yielding a mean pA2 value of 8.72. 4. In the continuous presence of 10(-6) mol/L enalaprilat, doxazosin with a pA2 value of 9.10 was a 2.4-fold more potent alpha 1-adrenoceptor antagonist than in the absence of enalaprilat. 5. These results are interpreted to indicate that endogenously produced angiotensin II can modulate the activity of alpha 1-adrenoceptors in vascular smooth muscle. PMID- 2568204 TI - Inhibition of transmitter release from sympathetic nerve endings by omega conotoxin. AB - 1. The effects of the calcium channel blocker, omega-conotoxin, on sympathetic neuroeffector function in the guinea-pig vas deferens have been investigated using a combination of mechanical and electrophysiological recording techniques. 2. Biphasic contractions evoked by electrical field stimulation were irreversibly abolished by omega-conotoxin (10-100 nmol/L). 3. Electrically evoked excitatory junction potentials and currents were irreversibly blocked by omega-conotoxin (10 100 nmol/L). Spontaneous excitatory junction potentials and currents were unaffected by this treatment. 4. omega-Conotoxin did not block impulse propagation in the nerve terminals. However, in three of four experiments omega conotoxin caused a decrease in the size of the nerve terminal impulse. 5. These findings support the suggestion the omega-conotoxin acts prejunctionally to inhibit sympathetic neuroeffector by interfering with depolarization-secretion coupling. PMID- 2568205 TI - Pathophysiology and management of unstable angina. AB - Significant progress has been made in recent years in unraveling the dynamic mechanisms involved in the production of unstable angina. This knowledge, and advances in medical and interventional therapy allow the formulation of treatment strategies aimed at specific pathogenic mechanisms and promise to reduce mortality and morbidity. This review covers the diagnosis, pathogenesis, risk stratification, and therapy of unstable angina. PMID- 2568206 TI - Adherence of vero cytotoxin-producing Escherichia coli serotype O157:H7 to isolated epithelial cells and brush border membranes in vitro: role of type 1 fimbriae (pili) as a bacterial adhesin expressed by strain CL-49. AB - Vero cytotoxin-producing Escherichia coli of the serotype O157-:H7 have recently been associated with sporadic cases and outbreaks of hemorrhagic colitis and with the hemolytic-uremic syndrome. E. coli O157:H7 strains adhere to human epithelial cells in tissue culture and to the intestine of orally infected rabbits. To determine whether E. coli O157:H7 adhere to normal post-natal human epithelial cells, we isolated buccal epithelial cells from healthy adult volunteers and isolated ileal enterocytes and colonocytes from histologically normal margins of surgical resection specimens. Apical brush border membranes from the distal ileum and colonocytes of the proximal colon were also prepared from post-weaning rabbits. Binding of five E. coli O157:H7 strains to epithelial cells and brush border membranes was determined by two complementary methods: firstly, under phase contrast microscopy and secondly, by quantitating adherence of radiolabelled bacteria to substrates that were bound to polystyrene microtiter wells. Under incubation conditions used previously to document in vitro adherence of other diarrheagenic E. coli, only the one type-1 fimbriated E. coli O157:H7 strain, designated CL-49, adhered to isolated human and rabbit epithelial cells. In contrast, binding of 4 non-type-1 fimbriated O157:H7 strains could not be demonstrated. Adherence of the fimbriated E. coli O157:H7 strain was saturable and varied with pH and temperature of the incubation medium. Adherence of bacteria to rabbit ileal brush borders was mediated by binding of bacteria to alpha-linked mannosyl residues present on surface glycoproteins.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568207 TI - [Clinical features of the peripheral nerve involvement in necrotizing angitis- characteristics in polyarteritis nodosa and allergic granulomatous angitis]. AB - Thirteen patients with peripheral neuropathy caused by necrotizing vasculitis were clinico-pathologically analyzed. These patients consisted of nine classical periarteritis nodosa (PN), four allergic granulomatous angitis (Churg-Strauss syndrome, AGA). All of them were proven to have a necrotizing vasculitis by sural nerve biopsy. The characteristics of peripheral neuropathy of these patients were summarized as follows. 1) Mononeuritis multiplex was a principal features in all patients preferentially localized in common peroneal, sural, radial median and ulnar nerves, with all modality of sensory impairment. 2) Radiation or diffuse deep-pain was a major initial symptom. Since this pain occurs frequently in the manner of sudden onset, the patient can tell the day of onset. 3) Local edema on the skin of involved region was initially observed. 4) Muscular atrophy and weakness was distributed more widely than sensory impairment. 5) Morphometric and teased-fiber study of biopsied sural nerves revealed axonal degeneration as a major pathological process. As compared to myelinated fibers, unmyelinated fibers were likely to be well preserved in morphology and population, which suggests that unmyelinated fibers are relatively resistant to ischemia. 6) Motor and sensory conduction study showed greatly decreased sensory and motor action potentials frequently resulting in absent of recordings. Conduction velocity is almost within normal range or just below the normal. Routine EMG recordings showed active denervation potentials in the involved muscles. 7) Protein in CSF was rarely elevated which suggested involvement of the spinal roots is infrequent. 8) Hypereosinophilia, thrombocythemia, fever, increased erythrocyte sedimentation rate, positive CRP and RA, and polyclonal hypergammaglobulinemia (IgG, IgA) were observed in most cases. PMID- 2568208 TI - [A case of tardive dystonia]. AB - Tardive dystonia in a chronic dystonia caused by neuroleptics. A 53-year-old man suffering from a neuroleptic induced dyskinesia began to show an abnormal posture. His abnormal posture was caused by changes of muscle tonus and thought to be a dystonic posture. He had no family history of dystonia. This posture was similar to that of idiopathic dystonia in that the muscle tonus was hypertonic in sitting but was hypotonic in lying, and in that the activity of daily living was not disturbed in spite of hypertonia. But he also showed lingual dyskinesia and hyperreflexia and the electromyographic analysis disclosed the fact that his dystonia was similar to that of secondary dystonia. Brain CT showed the atrophy of the head of caudate nucleus but superconducting MRI disclosed no abnormality in basal ganglia. No effective therapeutics was as yet found in tardive dystonia. So it was proposed that the tardive dystonia was due to irreversible functional damage to the basal ganglia. PMID- 2568209 TI - Population pharmacokinetic analysis of bisoprolol. AB - The technique of population pharmacokinetic analysis was employed to study the variability in the dose concentration relationship of bisoprolol during its clinical development. The influence of demographic factors on the variability of clearance was investigated in 3 different populations: group I, patients (including an elderly group) with essential hypertension receiving multiple oral doses of bisoprolol 10 or 20mg for 3 months; group II, patients with different degrees of renal impairment and healthy controls; and group III, patients with different types of hepatic impairment and healthy controls. Patients and controls in groups II and III received only a single oral dose of bisoprolol 10mg. The 3 data sets were analysed separately, using a non-linear mixed effects model (the NONMEM program). A 2-compartment pharmacokinetic model with first-order absorption described the data adequately. The typical values of volume of central compartment, volume of distribution at steady-state and the absorption rate constant for the 3 populations were: for group I, 68L, 235L, and 0.7h-1; for group II, 28L, 179L, and 0.3h-1; and for group III, 55L, 256L, and 0.4h-1, respectively. Plasma clearance was related to age in group I, to serum creatinine in group II and to aspartate transaminase activity in group III. The 68% confidence limits for clearance and elimination half-life were 8.2 to 21.5 L/h and 7.6 to 19.7h, respectively, for 50-year-old patients in group I. The analysis predicted that progressive increases in serum creatinine or aspartate transaminase activity will result in only a 50% reduction of clearance. PMID- 2568210 TI - Assessment of MK-912, an alpha 2-adrenoceptor antagonist, with use of intravenous clonidine. AB - MK-912, a new alpha 2-adrenoceptor antagonist, was assessed in six volunteers by use of antagonism of the effects of intravenous clonidine as the main index of response. Subjects received single doses of either 0.2 or 2 mg of orally administered MK-912 or placebo in a randomized, double-blind, balanced, crossover design. Clonidine was infused intravenously over 10 minutes, 1 hour after dosing, and observations were made for 8 hours. The 2 mg dose of MK-912 significantly inhibited the clonidine-induced hypotension, bradycardia, xerostomia, and increase in plasma glucose concentrations that were observed during the placebo treatment period (p less than 0.05). The peak elevation in plasma growth hormone that was produced by clonidine on the day the placebo was given was inhibited an average of 87% by the 2 mg dose of MK-912 (p less than 0.01). Although there was a trend toward antagonism of clonidine by the 0.2 mg dose of MK-912, statistically significant differences from placebo were not consistently demonstrated for most parameters. However, a mean 59% inhibition of the clonidine induced peak elevation of plasma growth hormone was observed (p less than 0.05). Oral MK-912 almost completely inhibits the effect of 200 micrograms of intravenous clonidine in human subjects, which is consistent with its role as a potent alpha 2-antagonist over the dose range of 0.2 to 2.0 mg. PMID- 2568212 TI - Nonsedating histamine H1-receptor antagonists. AB - The chemistry, pharmacology, pharmacokinetics, clinical efficacy, adverse effects, and dosages of the nonsedating histamine H1-receptor antagonists terfenadine, astemizole, loratadine, and acrivastine are reviewed. Terfenadine and astemizole are chemically unrelated to histamine H1-receptor antagonists such as diphenhydramine and chlorpheniramine. Loratadine is structurally related to the antihistamine azatadine, and acrivastine is a side-chain-reduced metabolite of the antihistamine triprolidine. Like other histamine H1-receptor antagonists, they competitively block histamine receptor sites rather than inhibiting histamine release. All four drugs have relatively long half-lives and are rapidly absorbed after oral administration. Terfenadine, astemizole, and loratadine are metabolized extensively in the liver; terfenadine and astemizole are both 97% protein bound. Terfenadine 60 mg twice daily has been shown to be as effective as conventional antihistamines for the treatment of seasonal allergic rhinitis. In clinical trials, astemizole 10 mg daily was comparable to or better than chlorpheniramine for treatment of chronic rhinitis. Both terfenadine and astemizole were effective for treatment of chronic urticaria. For treatment of seasonal allergic rhinitis, loratadine combined with pseudoephedrine may be preferable to triprolidine-pseudoephedrine and acrivastine-pseudoephedrine combinations that require more frequent dosing. Acrivastine must be administered more frequently than the other nonsedating antihistamines. None of these four agents impairs psychomotor activity. Infrequently reported adverse effects include dry mouth, skin reactions, and weight gain. The absence of substantial sedative effects and the less-frequent dosing schedules make these agents good alternatives to the classic antihistamines for treatment of seasonal and chronic rhinitis and chronic urticaria. PMID- 2568211 TI - Effects of dexmedetomidine, a selective alpha 2-adrenoceptor agonist, on hemodynamic control mechanisms. AB - Dexmedetomidine, a selective alpha 2-adrenoceptor agonist, was administered to five healthy male volunteers in single intravenous doses of 12.5, 25, 50, and 75 micrograms as part of a placebo-controlled study. The drug caused dose-dependent decreases in systolic and diastolic blood pressure. A small initial hypertensive response was observed after injection of the two highest doses. Heart rate was decreased. The concentration of norepinephrine in plasma was decreased significantly (by up to 92%), and the decrease was dose-dependent. No significant drug-induced alterations were observed in plasma renin activity or in the concentrations of atrial natriuretic peptide and arginine vasopressin in plasma. Other drug effects included dose-dependent impairment of vigilance and stimulation of growth hormone secretion. Plasma cortisol levels were unaffected. Dexmedetomidine is a potentially useful tool for studies of the physiology and pharmacology of alpha 2-adrenoceptors in human beings and may have therapeutic applications in clinical conditions in which sedative and sympatholytic effects are considered beneficial, such as premedication for anesthesia and surgery. PMID- 2568214 TI - Motor unit organization in developing muscle. PMID- 2568213 TI - Functional identification of 5HT receptor subtypes. PMID- 2568215 TI - Molecular analysis of the Duchenne muscular dystrophy locus. PMID- 2568216 TI - Neural mechanisms in disorders of movement. AB - 1. Experimental models of ballism, chorea and Parkinson's disease have been developed in the primate, and the underlying neural mechanisms which mediate these disorders of movement have been investigated using the 2-deoxyglucose uptake technique. 2. In ballism, the subthalamic nucleus is either lesioned or underactive. Because of the excitatory nature of subthalamic efferent fibres, this leads to abnormal underactivity of neurons in the medical segment of the globus pallidus which project to the ventral anterior and ventral lateral nuclei of the thalamus, and to the pedunculopontine nucleus of the caudal midbrain. 3. In chorea, there is underactivity of GABAergic striatal (putaminal) neurons which project to the lateral segment of the globus pallidus. This leads to overacting of lateral pallidal neurons and, thus, physiological inhibition of the subthalamic nucleus. Common neural mechanisms, therefore, underlie the appearance of dyskinesia in ballism and chorea. 4. In parkinsonism, there is overactivity of putaminal neurons projecting to the lateral pallidal segment. This results in excessive inhibition of lateral pallidal neurons and, as a consequence, disinhibition of the subthalamic nucleus. Overactivity of the subthalamic nucleus provides excessive drive upon medial pallidal neurons projecting to thalamic and pedunculopontine nuclei. PMID- 2568217 TI - The dorsal root reflex in isolated mammalian spinal cord. AB - 1. The dorsal root reflex has been investigated in an isolated preparation of adult mammalian spinal cord. 2. Both evoked and spontaneous activity can be recorded from the cord in the dorsal spinal roots. 3. The spontaneous activity has a characteristic pattern of firing in bursts of action potentials. Spontaneous and evoked activity are optimum at temperatures between 25 and 27 degrees C; little activity can be detected above 35 degrees C. 4. The spontaneous dorsal root activity has been shown to be correlated with negative potentials in the dorsal horn of the cord, and intracellular recordings made from primary afferent fibres have shown spontaneous primary afferent depolarizations (PAD) which underlie the generation of the spontaneous dorsal root activity. 5. The evoked dorsal root reflex has been shown to spread up to 16 spinal segments both rostrally and caudally from the stimulated dorsal root, and to the contralateral side of the cord. 6. The spontaneous dorsal root activity in widely separated segments has been shown by cross-correlation analysis to be linked both ipsi- and contra-laterally. 7. The significance of such a widespread system for the generation of PAD is discussed. PMID- 2568218 TI - Analysis of extracellular potentials using an IBM PC. AB - 1. A system has been developed for using IBM PC-compatible computers in combination with a Grafitek Data Logging Interface to record spike trains on magentic discs for later analysis. 2. The times and amplitudes of spikes detected on two input channels are recorded, together with a third channel containing information on computer-generated stimuli and keyboard-activated event markers. In excess of 50,000 spikes can be recorded with a computer having 640 k of Random Access Memory. 3. The recorded spike trains can be reconstructed on the computer monitor and keyboard-controlled window discriminators can be used to select the spikes for analysis by amplitude. 4. The same recorded data can be analysed to produce displays of spike count against time, amplitude histograms, inter-spike interval histograms, peri-stimulus time histograms(PSTH), raster displays and auto- and cross-correlations between activity on the two channels. Each spike is identified by number, allowing easy location of the start and finish of the section of data to be analysed, and the PSTH, raster and correlation analyses allow pretriggering to investigate event occurring before stimulation. 5. The axes of the displays histograms can be adjusted to produce optimum displays, and hard copy can be produced on dot matrix printers or digital plotters. 6. Quantitative analysis enables comparison between different recordings and treatments. PMID- 2568219 TI - Characterization of deep dorsal horn neurones in the rat spinal cord in vitro: synaptic and excitatory amino acid induced excitations. AB - 1. Two in vitro spinal cord preparations obtained from young rats (10-16 days), the transverse slice and the hemisected cord, have been utilized to examine the properties of deep dorsal horn neurones. 2. Several features have emerged: neurones respond to direct current injection with repetitive firing which is characteristically tonic in nature with little adaptation. Over the current intensities tested, no secondary firing range was apparent. 3. Graded afferent fibre stimulation produces a variety of sub- and suprathreshold postsynaptic excitatory potentials. The latencies of these potentials range from tens of milliseconds to hundreds of milliseconds, with the former predominating. 4. The majority of neurones are strongly excited by all three agonists: glutamate, quisqualate and N-methyl-D-aspartate but in addition a subpopulation of neurones with low sensitivity to glutamate and N-methyl-D-aspartate exists. 5. The implications of such properties for sensory processing within the dorsal horn are discussed. PMID- 2568220 TI - Amino acid pharmacology in a mature rat spinal cord preparation in vitro. AB - 1. It has been shown that a relatively large intact piece of mature mammalian spinal cord can be maintained in vitro if suitable experimental conditions are employed. 2. The preparation as described gives robust and reproduceable reflex responses in both dorsal and ventral horns (ventral horn activity has been maintained for over 36 hr in vitro). 3. The action of antagonists at both excitatory and inhibitory amino acid receptors give qualitatively predictable results from known in vivo experiments with the added advantage of fine quantitative control. 4. A significant advantage of such an adult preparation over more common immature spinal cord preparation is the ability to clearly separate low threshold reflexes, such as those described in this article, from reflexes evoked by small diameter non-myelinated afferents. PMID- 2568221 TI - Cytochemistry of the trigeminal and dorsal root ganglia and spinal cord of the rat. AB - 1. The primary sensory neurones have been classified into large light (LLC), type A, small dark (SDC), type B and type C cells on the basis of size, ultrastuctural and immunocytochemical characteristics. 2. Subclassifications have been described according to the configuration and spatial organization of cytoplasmic organelles. 3. Furthermore, the LLC are immunoreactive with a monoclonal antibody, RT97, directed against a neurofilament protein and the SDC are positive with anti-arginine vasopressin (AVP). 4. The majority of the neurochemical substances including substance P (SP), somatostatin (SOM), fluoride resistant acid phosphatase (FRAP), 5-hydroxytryptamine (5-HT) and glutamate were localized to the small and intermediate diameter neurones measuring 9-40 microns. 5. The cytochemistry of the dorsal horn was similar to the dorsal root ganglia (DRG). 6. There is good evidence that substance P (SP) and somatostatin (SOM) are transmitters for a proportion of nociceptive neurones but the neurotransmitters utilized by the rest of the subtypes are unknown. 7. 5-hydroxytryptamine (5-HT) and glutamate may be putative transmitters of the primary sensory neurones as they are localized in 28-30% of the SDC. 8. The wider distribution and extensive coexistence of the neuropeptides is incompatible with neurotransmitter function, but some may be neuromodulators whereas others such as arginine vasopressin (AVP) are useful markers for identifying type B neurones. PMID- 2568222 TI - The role of excitatory amino acids in synaptic transmission in the hippocampus. AB - We have highlighted some aspects of the action of excitatory amino acid transmission in the hippocampus. Fast epsps can be blocked by CNQX to reveal a component of synaptic transmission which is mediated by NMDA receptors. Extracellular recordings of ionic activities show that NMDA and non-NMDA ionophores are permeable to the major monovalent cations, while NMDA ionophores also appear to be permeable to Ca2+. Interactions of agonists applied by iontophoresis may be correlates of phenomena such as LTP, which can be evoked by appropriate synaptic stimulation. PMID- 2568223 TI - Hippocampal synaptic transmission: regulation of the NMDA receptor-mediated component. PMID- 2568224 TI - Function of synapses in the CA1 region of the hippocampus: their contribution to the generation or control of epileptiform activity. AB - 1. In the kainic acid lesioned hippocampus there is a loss of functional inhibition that is associated with reduction of the IPSPs recorded intracellularly from the surviving CA1 pyramidal cells. The possible pre- or postsynaptic origin of this change has been investigated. 2. Iontophoretic application of GABA to the soma and dendrites of CA1 pyramidal cells indicated that there had been no change in the efficacy of the postsynaptic GABA receptors on these cells. 3. Although a pre-synaptic mechanism is implicated, at one week post lesion we were unable to find any difference in the Ca+ dependent K+ evoked release of endogenous GABA. However, at survival times greater than 1 week immunohistological studies showed a decrease in the number of somatostatin positive non-pyramidal cells in the stratum oriens of the CA1 area. 4. In addition to the reduction of functional inhibition, changes in excitatory neurotransmitter mechanisms were also found to contribute to the epileptiform burst discharge. A slow component of the epileptiform EPSP recorded from CA1 pyramidal cells has been recorded and was found to be antagonized by the NMDA receptor antagonist D-APV. 5. Methods of controlling epileptiform activity in the kainic acid lesioned hippocampus have been tested. Stimulation of the substantia nigra and ventral tegmental areas produced profound inhibition of pyramidal cell activity in control hippocampi; however, they, were found to be ineffective in controlling the epileptiform burst. 6. A second method involved the use of hippocampal suspension grafts. Whilst this approach has yielded some encouraging data, further studies are necessary before the mechanism of the improvement in inhibitory synaptic function can be explained. PMID- 2568225 TI - The GABA system in functional tolerance and dependence following barbiturates, benzodiazepines or ethanol--correlation or causality? PMID- 2568226 TI - Inhibitors of brain GABA aminotransferase. PMID- 2568227 TI - Immunological approaches to the study of synaptic glycoproteins. PMID- 2568228 TI - The opiomelanocortin peptide family: neuronal expression and modulation of neural cellular development and regeneration in the central nervous system. AB - 1. Pro-OMLC is amongst a small number of propeptide-encoding genes which are expressed at highest levels in the CNS early in development. 2. The reappearance of the peptide products in injured neurones suggests that they fulfill a function in neuronal growth, differentiation and regeneration. 3. Axonal cues may regulate gene expression in neurones with greater or less functional interaction with their target cells. 4. alpha-MSH and ACTH stimulate the differentiation of neurones by accelerating their energy uptake and axonal growth during its early phases. 5. Their neurotrophic action is mediated through a common N-terminal amino acid sequence. 6. The structure activity requirements of the molecular second messenger responses underlying this action have yet to be conclusively determined. 7. Endorphins may regulate the transition from the mitotic cycle to the onset of differentiation of neurones and glia in the CNS. 8. Little is yet known of the cellular mechanism underlying this response, but the control of peptide processing to favour opiate and non-opiate receptor-mediated responses may be a key factor in determining whether they accelerate or retard neuronal differentiation. PMID- 2568230 TI - The effects of increasing pulmonary stretch and posture on sinus arrhythmia. AB - 1. It is generally accepted that sinus arrhythmia is due to change in vagal tone and that it is only present when vagal tone to the sino-atrial node predominates over sympathetic tone. Arguments persist about the origin of the feed-back information which initiates the arrhythmia, mostly among supporters of stretch receptors in the lungs and thorax (Anrep et al., 1936; Clynes, 1960), and baroreflexes in the blood vessels (Bainbridge, 1920; Davies and Neilson, 1967; Adeoshun, 1976). 2. The literature is scanty about the genesis of sinus arrhythmia. It would be worthwhile to investigate sinus arrhythmia under varying tidal volumes and posture. With this procedure it might be possible to differentiate between the importance of extrathoracic blood vessel stretch receptors and lung stretch receptors in the genesis of the arrhythmia. 3. This study is designed to investigate some of the causal factors involved in the production of sinus arrhythmia. PMID- 2568229 TI - Central cholinergic pathways and learning and memory processes: presynaptic aspects. PMID- 2568231 TI - Effects of soyabean and ascorbic acid on experimental carcinogenesis. AB - 1. Ultrastructural changes in liver tissue of mice fed nitrosamine precursors, dibutylamine and nitrite, were observed. 2. The protective effect of soyabean in a diet containing nitrosamine precursors was demonstrated. 3. Liver tissue was examined to investigate the anticarcinogenicity of ascorbic acid. 4. The significance of soyabean and ascrobic acid in counteracting the potential hazards due to nitrosamine precursors is discussed. PMID- 2568232 TI - Two measures of mental illness: contingent negative variation and spiral after effect. PMID- 2568233 TI - Physiological limitations to underwater exploration and work. PMID- 2568235 TI - The biology of the FMRFamide-series of peptides in molluscs with special reference to Helix. PMID- 2568234 TI - The physiological basis of transplantation of fetal catecholaminergic neurons in the transected spinal cord of the rat. AB - 1. Fetal (E.17) rat locus coeruleus and mesencephalic dopaminergic neurons when implanted into the transected spinal cord of the young adult rat survive for periods of longer than four months. Axons of up to 15 mm in length are observed growing from the cell bodies of the implanted neurons. 2. Fluorescent catecholaminergic (presumably dopaminergic) cell bodies are found in the caudal region of the transected, non-implanted spinal cord. 3. After transection of the spinal cord at the middle thoracic region in rats, at different postnatal ages (PN. 0, 7, 14, 21 and 28), there is substantial recovery of motor coordination involving all four limbs in the PN. 0 and PN. 7 groups. Recovery is best in the PN. 7 group. There is almost no recovery in the PN. 28 group, and very little recovery in the PN. 14 and PN. 21 groups. 4. Spinal locomotor generators in rat can, therefore, display a substantial degree of functional autonomy, if the spinal cord is cut before a certain critical stage of development (before PN. 14). These results have interesting implications with regard to current efforts to understand the mechanisms that regulate the spinal locomotor generators in experimental animals, and perhaps in man as well. PMID- 2568236 TI - Nicotinic-type unitary currents in Helix neurons. PMID- 2568237 TI - Use of an identified neurone in pesticide research. PMID- 2568238 TI - Frescon: a possible new pharmacological tool. PMID- 2568239 TI - Ionic currents in the soma of an identified cockroach motoneurone recorded under voltage-clamp. AB - 1. Membrane currents have been recorded from the soma of a bifunctional basalar/coxal depressor motoneurone in the metathoracic ganglion of the cockroach (Periplaneta americana) using a two-electrode voltage-clamp technique. 2. This motoneurone cell body is normally inexcitable when studied under current-clamp. Appropriate depolarizing command steps evoke rapid transient outward currents and late outward currents. 3. Late outward currents are dominated by a Ca-dependent component that confers an N-shaped I-V relationship on the neurone. 4. The Ca dependent outward current is suppressed by Cd2+ (1 mM), Mn2+ (5 mM) or verapamil (50 microM). 5. Externally applied tetraethylammonium ions (TEA+) (25 mM) block the Ca-dependent current, but also appear to suppress a component of the late outward current that is independent of Ca2+. 6. Aminopyridines cause only minor suppression of late outward currents, but shift the peak in the N-shaped I-V relationship to more negative potentials. 7. The reversal potential of tail currents recorded following pre-pulses to +50 mV were dependent upon the pre pulse duration; increasing the duration from 10 to 50 msec caused a +17 mV shift in tail current reversal potential. 8. A five-fold increase in the K+ concentration of the solution bathing the preparation only produced small and inconsistent changes in the reversal potential of tail currents. 9. Five-fold reduction in external Cl- caused no change. 10. The dependence of tail current reversal potential upon pre-pulse duration and the limited effect of alterations in the composition of the bathing solution are discussed in the context of restricted ion movements near the external surface of the cell membrane. PMID- 2568240 TI - Inactivation of calcium channels. AB - Rapid progress in our understanding of the properties and functions of voltage gated calcium channels had produced the need for an update to our previous review of calcium inactivation. The major elements of change included in this review are: 1. The existence of multiple forms of voltage-sensitive Ca+ channels, with distinctive single channel properties, thus necessitating a reappraisal of properties deduced from macroscopic current recordings, particularly of the processes of activation and inactivation. 2. The differences in biochemical properties between channel types are reflected in their differences in divalent selectivity, their requirement for metabolic maintenance and their mechanism of inactivation. These properties appear to divide the channels into two categories which may relate to their molecular structures. Further subgroupings, based upon the voltage thresholds, have also been observed. 3. Molecular properties of one class of channels have been elucidated, which correlate with the observed biochemistry of channel modulation and inactivation. 4. An enzymatic process underlying the mechanism of Ca2+-dependent inactivation has been elucidated and may serve as a model for other modulatory systems. The interweaving of the properties of these Ca2+ channels, with their spatial distributions and their influence upon other channel types, acts to transduce and integrate information within cells. PMID- 2568241 TI - Clinical experience with the use of laser radiation energy in the treatment of atherosclerosis. AB - The authors present their three-year experience with the use of their own method of laser angioplasty, called transluminal laser atherolysis, in 59 patients with various atherosclerotic vascular lesions. This low-intensity laser energy treatment was used alone or as adjunct to other endovascular surgical interventions. Early postoperative results were favourable in 82% of patients; in 20 patients followed-up for 2 years, the long-term results were satisfactory in 78%. The probable mechanisms involved in the beneficial effect of transluminal laser atherolysis are discussed. PMID- 2568242 TI - Some immunologic questions and predictions associated with the development of acquired immune deficiency syndrome. PMID- 2568243 TI - Restriction fragment length polymorphisms of the angiotensinogen gene in inbred rat strains and mapping of the gene on chromosome 19q. AB - Using a cDNA probe of the rat angiotensinogen gene (ANG), restriction fragment length polymorphisms (RFLPs) were detected in inbred rat strains with the restriction enzymes HindIII, PstI, and PvuII. Three alleles of ANG were almost equally distributed in 11 inbred strains. In two sets of backcross progeny originating from parental strains with different alleles, no close linkage was found between the ANG locus and 17 other loci tested. In situ hybridization, however, allowed assignment of the gene to chromosome 19q. The RFLPs of the angiotensinogen gene, therefore, can be considered useful as markers of rat chromosome 19. PMID- 2568245 TI - Lymphoid neoplasia and the control of haemopoietic differentiation. AB - Our broad aims are to delineate oncogenic events in lymphoid neoplasia and to search for genes that control haemopoietic differentiation. To explore lymphoid neoplasia, we have constructed transgenic mice bearing different oncogenes coupled to the immunoglobulin heavy chain enhancer (E mu), to force expression within lymphocytes. The prototype E mu-myc mice are highly prone to lymphomagenesis, generating pre-B and B cell lymphomas. In their pre-neoplastic phase, E mu-myc expression perturbs B cell development, accelerating the accumulation of pre-B cells. Lymphomagenesis requires additional oncogenic events, such as ras activation, and can be reconstructed in vitro. Transgenic mice bearing the N-myc, N-ras, v-abl and bcr-v-abl oncogenes are also prone to tumours. A striking demonstration that oncogenes can perturb lineage commitment has emerged. Introduction of the v-raf gene into cloned E mu-myc transgenic B cells frequently led to a switch in haemopoietic lineage: the cells became macrophages. Two clues to this remarkable metamorphosis are that the macrophage lines produce a myeloid growth factor and most bear marked karyotypic alterations, perhaps indicating that the balance between a few critical lineage control genes has been disturbed. To explore the hypothesis that genes encoding the DNA-binding homeo box domain participate in haemopoiesis, cDNA libraries from haemopoietic sources were screened, and several distinct homeo box cDNAs were isolated. They revealed a complex pattern of expression among haemopoietic cell lines. These genes are attractive candidates for regulators of haemopoietic differentiation. PMID- 2568244 TI - Chromosome specificity of satellite DNAs: short- and long-range organization of a diverged dimeric subset of human alpha satellite from chromosome 3. AB - The human alpha satellite DNA family, like many highly repeated satellite DNAs in eukaryotic genomes, is organized in distinct chromosome-specific subsets. As part of investigations into the molecular and evolutionary basis for the chromosome specific nature of such subsets, we report the isolation and characterization of alpha satellite sequences specific for human chromosome 3. This subset is characterized by a predominant tandemly arranged approximately 2.9 kb higher order repeat unit which, in turn, consists of 17 tandem diverged monomer repeat units of approximately 171 bp. Nucleotide sequence analysis reveals that the chromosome 3 higher-order repeat units are comprised, at least in part, of diverged dimeric (approximately 340 bp) sub-repeats and that this divergence accounts for the chromosome-specific behavior of this subset. Pulsed-field gel electrophoresis demonstrates that the chromosome 3 higher-order repeat units are localized in large domains, at least 1000 kb in length. Familial restriction fragment length polymorphisms associated with the satellite subset can be detected by pulsed-field gel electrophoresis and may facilitate molecular analysis of interchromosomal variation. PMID- 2568246 TI - Low bedtime doses of H2-receptor antagonists for acute treatment of duodenal ulcer. AB - Twenty-four-hour intragastric acidity was measured continuously over five separate occasions in 15 patients with healed duodenal ulcers. They were randomized to receive either placebo, cimetidine 800 mg, ranitidine 150 mg, famotidine 20 mg, or nizatidine 150 mg, given at 2200 hr in double-blind fashion. All H2-receptor blockers were more effective than placebo in suppressing both circadian (P less than 0.05-P less than 0.01) and nocturnal (P less than 0.002) gastric acidity, while there was no significant differences between the effects of the four active drugs in the same time periods. The percentage of nocturnal acid inhibition (2300-0800 hr) over placebo in terms of H+ values was virtually 100% with all active treatments. The effect on daytime (0800-1700 hr) and evening (1700-2300 hr) acidity of both placebo and the four H2-receptor antagonists was similar. Therefore, in the above doses H2-receptor blockers guarantee overnight anacidity to a similar degree and cause the physiological buffering of daily meals on gastric acidity to be fully exploited. Furthermore, the reducing effect of daily meals on drug action can be prevented. Since strong acid suppression strictly confined to the nocturnal period has been shown to be closely correlated with the highest ulcer healing rates, it is suggested that single low bedtime doses of H2-receptor antagonist should be evaluated in the acute treatment of duodenal ulcer. PMID- 2568247 TI - Randomized, double-blind comparison of famotidine with ranitidine in treatment of acute, benign gastric ulcer disease. Community-based study coupled with a patient registry. AB - A multicenter, double-blind, randomized controlled trial comparing the efficacy and safety of famotidine with ranitidine in the treatment of acute, benign gastric ulcer disease was coupled with a community-based gastric ulcer disease registry. One hundred ninety-five patients with endoscopically documented gastric ulcer disease were enrolled in the trial and randomly allocated to treatment with either famotidine 40 mg at bedtime or ranitidine 150 mg twice a day. Healing rates were similar in both groups: at four weeks 49% vs 48%, at six weeks 71% vs 69%, and at eight weeks 83% vs 81% for famotidine and for ranitidine, respectively. Pain relief, antacid tablet use, and adverse experiences were also similar in the two groups. Only 25% of patients entered in the gastric ulcer registry were enrolled in the trial. Given that patients with more severe or complicated gastric ulcer disease should be excluded from controlled trials of new drugs, the screening criteria used in the present study be excluded from findings being representative of a quarter of the patients seen in these practices. Therefore, coupling a patient registry with a clinical trial helps determine the applicability of its results. Famotidine 40 mg at bedtime is an effective and well-tolerated treatment of acute, benign gastric ulcer disease and is comparable in efficacy and safety to ranitidine 150 mg twice a day. PMID- 2568250 TI - Management of hypertensive emergencies. PMID- 2568248 TI - Gastrointestinal myoelectric activity in an infant with congenital idiopathic motility disorder. AB - We investigated myoelectric activity in an 8-month-old male who presented with a perinatal bowel obstruction, duodenal band, congenital short small intestine, and persistent feeding intolerance. Serosal electrodes were surgically implanted on stomach, duodenum, and jejunum during Nissen fundoplication and ileostomy. A 5-cm ileal specimen was taken for in vitro studies. Spontaneous migrating myoelectric complexes (MMC) were present in stomach and small intestine. Bethanechol increased electrical response activity (ERA) in stomach and duodenum. Morphine induced intense ERA and distinct phase III activity. Pentagastrin infusion did not disrupt MMC cycling. Feeding disrupted MMC complex cycling 30-40 min after the meal. Metoclopramide before feeding delayed disruption of the MMC cycling after the feeding. Intermittent gastric arrhythmias were present after the fifth postoperative day. In vitro muscle strips showed spontaneous contractions and electrical control activity (ECA). Bethanechol, McNeil A-343, motilin, and cholecystokinin induced contractions, but pentagastrin had no effect. We conclude that in spite of a major clinical motility dysfunction, several of our findings were normal. The abnormalities include short MMC period, absence of disruption of MMC by pentagastrin, and gastric arrhythmias. PMID- 2568249 TI - Immunocytochemical staining of astrocytes, tanycytes, immature glial cells and ependymal cells in the central nervous system of the normal rat and the myelin deficient mutant. AB - Central nervous system (CNS) tissue from young myelin-deficient (md) mutant rats and male littermate controls was immunostained with antibodies against the following glial-cell antigens: glial fibrillary acidic protein (GFAP), glutamine synthetase, vimentin, the Yb isozyme of glutathione-S-transferase (Yb) and carbonic anhydrase. Gliosis was most severe in spinal-cord 'white-matter' tracts in the md rat. The most dramatic differences between astrocytes in the md and normal animals were revealed with anti-GFAP and anti-vimentin. Ependymal cells and tanycytes were quite similar in the md and normal CNS. Among the glial-cell antigens, Yb showed the broadest distribution, which included astrocytes, ependymal cells and tanycytes. PMID- 2568251 TI - [Experimental studies on the effect of Bulgarian anxiolytic lonetil on some endocrine functions]. PMID- 2568252 TI - [Involvement of glutamate released from cortico-striatal fibers in the presynaptic control of dopamine liberation in the caudal nucleus of the cat]. PMID- 2568253 TI - [Use of the methylxanthines, caffeine and theophylline, in the treatment of extrapyramidal disorders caused by treatment with neuroleptics. Development of a therapeutic hypothesis]. PMID- 2568254 TI - [Symposium. Neurotransmitter interactions: implications in neuro and psychopathology. Dopaminergic systems. Cannes, 4-6 March 1988. Proceedings]. PMID- 2568255 TI - [Ascending mesencephalic dopaminergic pathways in the rat. Immunohistochemical demonstration of the coexistence of the amine with CCK-8 or neurotensin]. PMID- 2568256 TI - Urinary output modulation of alanine aminopeptidase, gamma-glutamyl transpeptidase and N-acetyl-beta-D-glucosaminidase by castration and testosterone in male normal rat. AB - The urinary excretion of alanine aminopeptidase (AAP), gamma-glutamyl transpeptidase (gamma-GT) and of N-acetyl-beta-D-glucosaminidase (NAG) was studies in normal and castrated rats receiving either testosterone for 5 post operative weeks or no hormone. In castrated rats the urinary output of AAP and gamma-GT was significantly lower than in sham control or in castrated rats receiving testosterone. In addition, an excess of exogenously given testosterone had no effect on enzymuria of normal rats. The urinary excretion of NAG was influenced neither by castration nor by testosterone. These results suggest that endogenous testosterone is responsible for a permanent positive control on the urinary excretion of AAP and gamma-GT. PMID- 2568257 TI - Mitogen activation of human lymphocytes from normal and acute lymphocytic leukemia (ALL) subjects. AB - Lymphocytes from acute lymphocytic leukemia (ALL) subjects were converted by mitogens to blast-like cells whose microscopic appearance and rate of formation was indistinguishable from those in mitogen incubated control lymphocytes. In ALL lymphocytes, however, pokeweed mitogen (PWM) failed to stimulate GGT expression; the mean increase it caused in thymidine kinase (TK) activity and thymidine incorporation was normal, though there were appreciable individual variations. These variations were also apparent with concanavalin A (Con A) but, in most ALL cases, TK and thymidine incorporation rose to much higher levels than in Con-A treated control lymphocytes. The results indicate that evaluation of the response to mitogens by quantitative biochemical criteria provides a sensitive method for revealing functional impairments in microscopically normal ALL lymphocytes. PMID- 2568259 TI - Presence of a lysosomal enzyme, arylsulfatase-A, in the prelysosome-endosome compartments of human cultured fibroblasts. AB - Although endosomes and lysosomes are associated with different subcellular functions, we present evidence that a lysosomal enzyme, arylsulfatase-A, is present in prelysosomal vesicles which constitute part of the endosomal compartment. When human cultured fibroblasts were subfractionated with Percoll gradients, arylsulfatase-A activity was enriched in three subcellular fractions: dense lysosomes, light lysosomes, and light membranous vesicles. Pulsing the cells for 1 to 10 min with the fluid-phase endocytic marker, horseradish peroxidase, showed that endosomes enriched with the marker were distributed partly in the light lysosome fraction but mainly in the light membranous fraction. By pulsing the fibroblasts for 10 min with horseradish peroxidase conjugated to colloidal gold and then staining the light membranous and light lysosomal fractions for arylsulfatase-A activity with a specific cytochemical technique, the endocytic marker was detected under the electron microscope in the same vesicles as the lysosomal enzyme. The origin of the lysosomal enzyme in this endosomal compartment was shown not to be acquired through mannose 6-phosphate receptor-mediated endocytosis of enzymes previously secreted from the cell. Together with our recent finding that the light membranous fraction contains prelysosomes distinct from bona fide lysosomes and was highly enriched with newly synthesized arylsulfatase-A molecules, these results demonstrate that prelysosomes also constitute part of the endosomal compartment to which intracellular lysosomal enzymes are targeted. PMID- 2568258 TI - Autoregulation and multiple DNA interactions by a transcriptional regulatory protein in E. coli pili biogenesis. AB - An operon mediating biogenesis of digalactoside-binding pilus-adhesin of serotype F13 in uropathogenic Escherichia coli includes the regulatory gene papB. The papB gene product was found to act as transcriptional activator of an operon which includes the papB gene and several pap cistrons encoding the proteins of the pilus polymer. Studies of how pap gene expression was affected by increasing amounts of PapB protein in the cells showed that high levels did not stimulate transcription but caused repression. Results from in vitro studies demonstrated that the PapB protein was a sequence-specific DNA-binding protein. Binding studies using gel mobility shift assays and DNase I protection (footprinting) showed that PapB protein binds to three separate sites. A sequence greater than 200 bp upstream of the promoter, and directly adjacent to a binding site for the cAMP receptor protein-cAMP complex, appeared as a preferential PapB binding site. A second site was localized to sequences overlapping the -10 region of the promoter and a third binding site was found within the coding sequence of the papB gene itself. The data suggest that the PapB protein has a dual function as activator/repressor of pilus-adhesin transcription and that its autoregulatory mode of action involves differential binding to separate sites. PMID- 2568260 TI - Microtubule and centrosome distribution during sheep fertilization. AB - The distribution of microtubules and centrosomes was studied during sheep fertilization by electron and immunofluorescence microscopy. Tubulin and centrosomal material was identified with monoclonal anti-alpha-tubulin and MPM-2 antibodies, respectively. In ovulated eggs, microtubules were exclusively found in the meiotic spindle and centrosomal material at each of its poles. At fertilization, sperm centrosomes were incorporated into the egg and organized the sperm astral microtubules. During pronuclear development and migration, the sperm aster increased in size; microtubules of the sperm aster extended from the male pronucleus to the egg center and towards the female pronucleus. The position of the sperm aster during pronuclear migration suggests that it plays a role in this process. When the pronuclei were in apposition in the egg center, a dense array of microtubules and the centrosomal material were present between the two pronuclei. The proximal centriole of the sperm was identified by electron microscopy, between the apposed pronuclei. The centrosomal material extending around the centriole and the sperm neck and proximal mid-piece, apparently contained several foci from which microtubules radiated. These data suggest that in sheep unlike in mice, centrosomal material originating from the sperm is involved in the fertilization events. PMID- 2568262 TI - Biochemical evidence for somatostatin receptors in murine neuroblastoma clone N1E 115. AB - Radioligand binding and functional assays were employed to demonstrate the existence of somatostatin receptors in the murine neuroblastoma clone N1E-115. Saturation experiments with [125I][Tyr11]somatostatin-14 indicated the presence of a single class of binding sites in membranes prepared from N1E-115 cells (Kd = 83 pM; Bmax = 21,000 receptors/cell). Somatostatin-14, somatostatin-28 and L363586 (cyclo(N-Me-ALA-TYR-D-TRP-LYS-VAL-PHE] all displaced the 125I-ligand monophasically in N1E-115 cells (Ki values were 28, 82 and 34 pM, respectively), which contrasted with the binding heterogeneity apparent with L363586 in rat brain membranes. The binding of [125I][Tyr11]somatostatin-14 was reduced by GppNHp, indicating that N1E-115 somatostatin receptors interacted with guanine nucleotide binding protein(s). Somatostatin agonists decreased by 30-50% the levels of [3H]cyclic AMP induced in intact cells by forskolin, prostaglandin E1, or vasoactive intestinal polypeptide. The EC50 values for inhibition of the [3H]cyclic AMP response to PGE1 by L363586, somatostatin-14, and somatostatin-28 were 0.24, 0.63 and 1.0 nM, respectively. Pertussis toxin treatment of N1E-115 cells reduced both binding to the receptor and the functional response to somatostatin-14. These data suggest that a single class of somatostatin receptors in N1E-115 cells are linked to the inhibition of adenylate cyclase through a Gi protein. PMID- 2568261 TI - Effect of bisoprolol on cardiac performance in coronary heart disease. AB - The effects of 5 and 10 mg bisoprolol once daily for 7 days on exercise ECG, myocardial perfusion and left ventricular function in 25 patients with stable coronary heart disease have been assessed in a double-blind, randomized, parallel group trial design. ST-segment depression during exercise was reduced by 56% by 5 mg bisoprolol and by 64% after 10 mg; the difference between the dose levels was significant. Heart rate, systolic and diastolic blood pressure and the rate pressure product were reduced to similar extent both at rest and during exercise by both doses. Left ventricular thallium-201 scintigrams indicated a significant reduction in myocardial perfusion defects after 10 mg bisoprolol compared to baseline; however, the difference between the two active treatments was not significant. Left atrial and left ventricular diameters obtained by one dimensional echocardiography, and the calculated shortening fraction, remained unchanged after bisoprolol, and so gave no evidence of a negative inotropic action. It is concluded that 5 mg bisoprolol was effective in once-a-day treatment of angina pectoris due to coronary heart disease, and a further improvement can be expected on increasing the dose to 10 mg. PMID- 2568263 TI - [3H]SCH 23390 labels a novel 5-hydroxytryptamine binding site in human blood platelet membranes. AB - In human blood platelet membranes, 5-HT displaced the binding of the putative selective D-1 dopamine receptor antagonist [3H]SCH 23390 in a competitive manner with a Ki value of 5.7 +/- 0.8 nM, which was about 1000-fold lower than the Ki value for dopamine (Ki = 4400 +/- 150 nM). Thus the 'D-1 dopamine-like' site in human blood platelet membranes described previously corresponds to a 5-HT1-type site. [3H]SCH 23390 competition experiments with a number of serotonergic drugs disclosed a pharmacological profile that was distinct from the four 5-HT1 site subtypes reported previously. We therefore propose that this novel 5-HT site be designated the 5-HT1E site. Binding of [3H]SCH 23390 to 5-HT1-type sites could not be detected in several regions of the human brain. In some regions, however, 5-HT displaced part of the [3H]SCH 23390 binding with a K1 value of 320-380 nM. These sites correspond to 5-HT2 receptors. PMID- 2568264 TI - Glycine and kynurenate modulate the glutamate receptors in the myenteric plexus and in cortical membranes. AB - The responses evoked by stimulation of the N-methyl-D-aspartate receptors in the guinea-pig myenteric plexus were potentiated by micromolar concentrations of glycine and were non-competitively antagonized by kynurenate (IC50: 60 microM). The effects of kynurenate were competitively prevented by glycine. Furthermore, kynurenate displaced [3H]glycine from its binding sites on rat cortical membranes (IC50: 20 microM). Kynurenate and glycine, therefore, probably act at the same site, evoking opposite effects on the function of the ion channel complex of the N-methyl-D-aspartate receptor. PMID- 2568265 TI - Selective sigma receptor agonist and antagonist affect dopamine neuronal activity. AB - Extracellular recording techniques were used to study the effects of the selective sigma receptor agonist (+)-[3H]3-(3-hydroxyphenyl)-N-(1 propyl)piperidine ((+)-3-PPP) and selective sigma receptor antagonist BMY 14802 on dopamine (DA) neurons of the substantia nigra. Intravenous administration of (+)-3-PPP produced a dose-dependent inhibition of DA neuron firing rate. Complete inhibition of DA neurons produced by (+)-3-PPP could be completely reversed by administration of BMY 14802. Also, pretreatment with BMY 14802 shifted the (+)-3 PPP dose response curve to the right. These data demonstrate a relationship of the sigma receptor with the dopamine system and further suggest a model system to study agonist/antagonist interactions of sigma ligands. PMID- 2568266 TI - MIF-1 attenuates apomorphine stereotypies in adult rats after neonatal 6 hydroxydopamine. AB - Since prolyl-leucyl-glycinamide (MIF-1) modifies the behavior of adult rats after treatment with neuroleptics, we examined whether MIF-1 would also modify adult behavior after treatment of neonatal rats with 6-hydroxydopamine (6-OHDA). Rats received 6-OHDA (100 micrograms i.c.v.) or diluent at 3 days after birth and either MIF-1 (2.0 mg/kg per day s.c. x 10 days) or diluent beginning at 28 or 29 days after birth. At 5 weeks, a low dose (0.1 mg/kg s.c.) of apomorphine increased the distance traveled, time in ambulation, number of stereotypic movements, and number of movements per time in stereotypy, but decreased the time in stereotypy in the 6-OHDA group. MIF-1 (x 7 or 8 days) showed a tendency to attenuate the increased number of movements and significantly (P less than 0.05) reduced all of the other effects of neonatal 6-OHDA. Behavior induced by higher doses of apomorphine in the 6-OHDA group (reduced licking and head nodding; increased paw treading, taffy pulling and self-biting) were not attenuated by MIF 1. At 38 or 39 days, total in vitro binding of [3H]SCH-23390 and [3H]spiroperidol to striatal homogenates was not altered in any of the groups. The findings demonstrate that specific early developmental alterations in apomorphine-induced behaviors can be modified by treatment of adult rats with MIF-1, even in the absence of overt changes in the binding of striatal dopamine D-1 and D-2 receptors. PMID- 2568267 TI - Nigrostriatal reconstruction after 6-OHDA lesions in rats: combination of dopamine-rich nigral grafts and nigrostriatal "bridge" grafts. AB - In an attempt to reconstruct the nigrostriatal dopamine (DA) pathway following 6 hydroxydopamine-induced degeneration in adult rats, a novel double graft procedure has been developed. Embryonic DA-rich grafts were implanted in the vicinity of the host substantia nigra by standard procedures. An intracerebral bridge was then implanted by injection of alternative tissues along a single oblique needle penetration through the frontal pole and neostriatum to the substantia nigra. Embryonic striatal tissue bridges provided a continuous column of tissue from the nigral DA graft to the host striatum. In 3 cases, tyrosine hydroxylase immunoreactive fibres grew the full length of the striatal bridge grafts and were seen to penetrate the host neostriatum. In these three animals alone, methylamphetamine-induced rotation was significantly reduced to 44% of the baseline lesion-induced turning rate. Alternative tissues or substrates (embryonic olfactory bulb, cultured astrocytes or laminin-coated microspheres) were not effective in promoting THir fibre growth from the nigral DA grafts to the host neostriatum. PMID- 2568268 TI - Extracellular concentrations of aspartate and glutamate in rat neostriatum following chemical stimulation of frontal cortex. AB - Extracellular concentrations of excitatory amino acids in the neostriatum of anaesthetized rats were studied by in vivo microdialysis and the influence of chemical stimulation of the neocortex assessed. Administration of gamma aminobutyric acid (GABA) antagonists to the neocortex evoked an increase in the extracellular concentration of aspartate (to 315 and 210% of basal values for microinfusion and topical application, respectively), but not of extracellular glutamate. Such selectivity was also found in response to N-methyl-D-aspartate (NMDA, 348% increase in extracellular aspartate concentration). Yet co administration of NMDA with GABA antagonists increased the extracellular concentration of glutamate (to 278% of basal values) as well as aspartate (to 611% of basal values). The results suggest that either the activated neurons are hypoglycemic or hypoxic, or that aspartate is the major neurotransmitter of corticostriatal neurons. PMID- 2568270 TI - Neurochemical heterogeneity among corticofugal and callosal projections. AB - Biochemical, physiological, and anatomical studies over the past 30 years have firmly established glutamate (Glu) as the major neurotransmitter of those cortical neurons which give rise to corticofugal pathways. In the present study we utilized immunohistochemistry, with an antibody directed against Glu, in conjunction with wheat germ agglutinin-horseradish peroxidase (WGA-HRP) histochemistry to examine the Glu-containing neurons which give rise to corticofugal and callosal projections of the rat. Injections of WGA-HRP into the pons labeled cells in layer V of both visual and somatosensory cortices. WGA-HRP labeled cells which also stained for Glu were large pyramids and in the visual cortex constituted approximately 42% of the total number of neurons which had effectively transported WGA-HRP, while the percentage was 56% in the somatosensory cortex. Following caudate/putamen injections, WGA-HRP-labeled cells were confined to layer V of the somatosensory and motor cortices. Of these cells, 40% in the somatosensory cortex and 53% in the motor cortex were also stained for Glu. Finally, after WGA-HRP injections in the visual cortex numerous WGA-HRP positive neurons were found throughout layers II-VI around the boundaries between area 17 and areas 18 and 18a of the contralateral hemisphere. Here, 38% of these cells were also labeled for Glu, but this percentage was higher (49%) when layers II-III were considered alone. These findings show that Glu is not the neurotransmitter used overwhelmingly in corticofugal and callosal projections and that different proportions of neurons are Glu-immunoreactive in the systems examined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568269 TI - 3-Acetylpyridine-induced degeneration of the nigrostriatal dopamine system: an animal model of olivopontocerebellar atrophy-associated parkinsonism. AB - The effects of 3-acetylpyridine (3-AP) administration to rats on the mesotelencephalic dopamine system were assessed. A single 3-AP injection resulted in biochemical and immunohistochemical evidence of degeneration of the nigrostriatal dopamine system. Six weeks after 3-AP treatment decreases in both striatal dopamine content and the activity of the catecholamine biosynthetic enzyme tyrosine hydroxylase were observed. Immunohistochemical examination suggested a decreased density of striatal tyrosine hydroxylase-immunoreactive fibers and revealed the emergence of a distinctly patchy organization of the dopamine innervation to the dorsolateral striatum. While 3-AP administration resulted in biochemical and anatomical data consistent with the degeneration of nigrostriatal dopamine fibers, no significant changes in dopamine content or the density or pattern of tyrosine hydroxylase-immunoreactive fibers in the anteromedial prefrontal cortex or nucleus accumbens were seen. These data suggest that 3-AP administration may result in a relatively specific degeneration of the nigrostriatal dopamine system. Since 3-AP causes both a profound loss of the climbing fiber input to the cerebellum derived from the inferior olivary nucleus, and the degeneration of nigrostriatal dopamine neurons, 3-AP administration may provide a useful model of olivopontocerebellar atrophy-associated parkinsonism. Moreover, the differences in the neurotoxicity caused by 3-AP and that elicited by another pyridine which causes striatal dopamine depletion (1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine, MPTP) may offer important insights into the mechanisms of both species- and site-specific pyridine neurotoxins. PMID- 2568271 TI - [Effect of quinpirole on the retrieval of a memory trace in amnesia]. AB - In experimental studies on mice it was shown that quinpirole, a selective agonist of dopaminergic postsynaptic D-2 receptors, possessed a pronounced antiamnestic activity. Administration of quinpirole before testing on 2nd and 22nd days reactivated the retrieval of passive avoidance impaired by an amnestic agent. The drug effect depended on the duration of "psychogenic" amnesia. It is suggested that the postsynaptic dopaminergic D-2 receptors are actively involved in the neurochemical mechanisms of memory trace retrieval. PMID- 2568272 TI - [The pharmacology of 1-(2-pyrimidinyl)piperazine, a metabolite of buspirone]. AB - In experiments on rats the anxiolytic activity of 1-(2-pyrimidinyl)-piperazine (PP) was revealed only in some models of anxiety states, although diazepam was effective in all used models. In contrast to diazepam, PP exerts the anesthetic potentiating and myorelaxant effects only when administered in subtoxic doses and possesses no anticorazole activity. The facts testifying to the serotoninergic mechanisms of the anxiolytic action of 1-(2-pyrimidinyl)-piperazine are presented. PMID- 2568273 TI - [Effect of calcium-channel blockers on human thrombocyte aggregation induced by alpha-adrenoreceptor agonists]. AB - A comparative study of the effects of calcium channel blockers nifedipine and verapamil on platelet aggregation in healthy donors induced by adrenaline. A partial agonist of alpha 2-adrenoreceptors clofelin and also clofelin in combination with the threshold concentrations of ADP inducing aggregation was performed. It was found that verapamil is a more potent inhibitor of platelet aggregation induced by adrenaline as compared with nifedipine inhibiting this process only at high concentrations. The aggregation-inducing effect of a partial agonist of alpha 2-adrenoreceptors clofelin is eliminated at low concentrations in the aggregating mixture of nifedipine than verapamil. PMID- 2568274 TI - [A rapid radioenzyme method for determining dopamine and 3,4 dihydroxyphenylacetic acid in the basal ganglia of the rat brain]. AB - A rapid and high sensitive radioenzymatic method of determination in the rat brain basal ganglia of dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DHPA) whose methylated derivatives were divided by extraction with organic solvents is proposed. The method sensitivity for DA is 0.25 ng in a sample and for DHPA 0.1 ng in a sample. The yield of the internal standard is 80-85%. The method is high specific, the cross reaction during determination of DA and DHPA does not exceed 5%. The study of the effects of some dopaminergic agents (apomorphine, haloperidol. L-DOPA and pargyline on the contents of DA and DHPA in the rat brain basal ganglia in vivo confirms a high specificity of the method. The method may be used for screening of pharmacological compounds possessing the suggested dopaminergic and antimonoamine oxidase activity. PMID- 2568275 TI - [Hemorrhagic fever with renal syndrome]. PMID- 2568276 TI - [The role of the paramedical worker in preventing allergic diseases in children]. PMID- 2568277 TI - Salivary testosterone in cryptorchid pubertal boys: a parameter to assess the gonadal function. AB - The authors studied salivary and serum testosterone (T) levels in basal conditions and at 6 and 12 months after orchidopexy in 36 pubertal boys with unilateral or bilateral cryptorchidism. Boys were divided into two groups according to Tanner classification. Salivary and serum T levels were significantly lower in the group with scanty or absent sexual development than those obtained in the group with normal development. Both groups exhibited similar saliva and serum testosterone values at 12 months of the follow-up study. The data shown indicate that salivary T seems a good index for evaluating the androgenic status of these patients. PMID- 2568279 TI - [Effects of neonatal administration of monosodium glutamate on four neuropeptide concentrations in the rat brain]. AB - The present study was designed to measure concentrations of four neuropeptides in different brain regions in monosodium glutamate(MSG)-treated rats and to assess molecular forms of each peptide with gel and high performance liquid chromatography (HPLC). MSG(4mg/kg body weight) or 10% NaCl was injected subcutaneously on postnatal days 1, 3, 5, 7 and 9 to male littermates which were subsequently used on postnatal day 100. Rats were sacrificed by decapitation, and the brains were dissected into ten discrete regions. The brain extracts were subjected to measurement of four neuropeptides; somatostatin (SRIF), neuropeptide Y (NPY), atrial natriuretic polypeptide(ANP), and a novel pituitary polypeptide 7B2 by specific radioimmunoassays. Significant increase (p less than 0.01) in midbrain SRIF content was observed in MSG-treated rats, though there was no significant change in hypothalamic SRIF content. Significant reduction (p less than 0.05) in hypothalamic NPY content was also found in MSG-treated rats. Hypothalamic ANP content was similar in both MSG-treated and control rats. A significant increase of 7B2 content was found in substantia nigra/ventral tegmentum and hypothalamus (p less than 0.05 or p less than 0.01, respectively) in MSG-treated rats. These four immunoreactivities were further characterized by gel permeation or high pressure liquid chromatography (HPLC). Chromatographic analysis of SRIF immunoreactivity revealed that there were two distinctive peaks and smaller molecular weight component corresponding to SRIF. Fractionation of NPY or 7B2 immunoreactivity by gel permeation showed a single major peak which was identical to the synthetic NPY or 7B2 immunoreactivity from porcine pituitary extract. HPLC analysis for ANP immunoreactivity also showed that the major immunoreactive component corresponded to synthetic rat ANP. MSG treatment could not produce any major alterations in proportions of molecular forms studied. These results suggest that MSG treatment in neonates might produce the alterations in SRIF, NPY and 7B2 content in the discrete brain regions including the hypothalamus. PMID- 2568278 TI - [The effects of PGE2, 2-DG and L-arginine on hypothalamic GHRH and SRIF releases in conscious rats, with respect to GH secretion]. AB - The effects of prostaglandin E2 (PGE2), 2-deoxy-D-glucose (2-DG) and L-arginine on hypothalamic GHRH and SRIF release with respect to GH secretion were studied in conscious male rats. Intracerebroventricular (icv) injection of 5 micrograms PGE2 and intravenous (iv) infusion of 1 g/kgBW L-arginine caused an increase in plasma GH levels, but icv (36 micrograms) or iv (400 mg/kgBW) injection of 2-DG suppressed spontaneous GH surge in conscious rats. The concentration of hypothalamic GHRH was decreased in all three groups of the animals, but the concentration of hypothalamic SRIF was decreased only in 2-DG-treated animals. In the perifusion system using rat hypothalamus, PGE2 (0.28 microM, 2.8 microM), 2 DG (22 mM) and L-arginine (3 mM) stimulated GHRH release from rat hypothalamus. 2 DG also stimulated SRIF release more predominantly than GHRH release. Passive immunization with anti-GHRH serum inhibited the GH secretion induced by icv injection of 5 micrograms PGE2 and by iv infusion of 1 g/kgBW L-arginine in conscious rats. In contrast, GH secretion induced by iv injection of 50 micrograms/kgBW PGE2 was not affected by the pretreatment with the antiserum. These results suggest that the central effect of PGE2 and peripheral effect of L arginine to stimulate GH secretion are mediated by hypothalamic GHRH release, and that the inhibitory effect of 2-DG on GH secretion is predominantly mediated by hypothalamic SRIF release rather than GHRH release in rats. PMID- 2568280 TI - Some observations on conserved polar side chains in immunoglobulin V-domains. AB - 1. The roles of conserved polar residues have been studied in 12 V-domains for which atomic coordinates are available. 2. In most cases a particular residue had a similar side chain conformation in all V-domains examined and the polar group provided the same hydrogen bonds which helped to stabilize the conformations of the domains. 3. In the case of a conserved glutamine/glutamic acid residue the buried side chain could adopt a variety of conformations and the polar group could form different hydrogen bonds from one domain to another. However, they contributed similarly to domain stability. 4. In the case of a conserved threonine/serine residue its side chain showed relative rotations of up to 180 degrees from one domain to another. The hydroxyl group could be buried or exposed at the domain surface. In some domains it formed hydrogen bonds to two other protein atoms but in other domains there was a single hydrogen bond or none at all. The varied roles of this residue are discussed in the text. PMID- 2568281 TI - Calmodulin-stimulated particulate guanylate cyclase in crayfish hepatopancreas. AB - 1. A particulate guanylate cyclase from crayfish hepatopancreas membranes was investigated with respect to its dependence on Ca2+ and calmodulin. Addition of Ca2+ to EGTA-treated membranes increased cyclase activity by 100%. 2. Calmodulin stimulated the activity about 5-fold. 3. This effect could be abolished by the calmodulin antagonist compound 48/80. 4. These results present evidence that the particulate guanylate cyclase of crayfish hepatopancreas is a Ca2+/calmodulin dependent enzyme. 5. The implications of this observation upon glycogen metabolism of crustaceans are discussed. PMID- 2568282 TI - Safe motherhood: an international priority. PMID- 2568283 TI - Chlamydia trachomatis infections. AB - The diversity in chlamydial syndromes and their consequences especially the impact on human reproduction and fertility, the asymptomatic characteristics of infections and the ever-increasing incidence have been the focus of considerable attention in the 1980s. The current status of the clinical spectrum of ocular genital infections due to Chlamydia trachomatis and the laboratory tests for accurate, rapid diagnosis and effective treatment are reviewed. PMID- 2568285 TI - Ultrasonic prediction of growth discordancy by intertwin difference in abdominal circumference. AB - Intertwin abdominal circumference (AC) and femur length (FL) differences were studied in 24 growth-discordant (greater than or equal to 15% birth weight difference) and in 32-growth concordant twin pairs delivered within 2 weeks of the last ultrasonic examination. Both groups were of similar gestational age but differed significantly in the mean twin birth weight and intertwin AC difference (P = 0.01 and P = 0.00009, respectively). A cutoff value (greater than or equal to 18 mm) for AC difference was found to discriminate significantly (P less than 0.0009) between concordant and discordant pairs with sensitivity, specificity, positive predictive value and negative predictive value of 66.7%, 78%, 69.5% and 75.7%, respectively. These data may suggest that an intertwin difference of 18 mm or more in AC effectively screens for the diagnosis of 15% or more birth weight difference in twin pregnancies. PMID- 2568284 TI - Severe abruptio placentae--still unpreventable. AB - The management of abruptio placentae remains a problem, despite advances in medical science. This study analyses severity of abruption, resultant morbidity, and maternal and fetal mortality in 105 cases. Among the contributory factors, hypertension (44%) and grand multiparity (24%) were common. There was no maternal death among these cases, mainly because of liberal use of blood transfusion, prompt delivery, and careful fetal monitoring, but perinatal mortality remained high (73%). Factors responsible for this high perinatal mortality and its prevention are discussed. PMID- 2568286 TI - Effect of fetal sex and race on amniotic fluid lecithin concentration. AB - Amniotic fluid lecithin phosphorus concentration (AF-Lec) was measured in 209 healthy women at 31.7-42.7 weeks gestation. The patients were divided into four groups according to race and fetal sex. No differences in AF-Lec between black and white nor between male and female fetuses were found. The relationship of AF Lec to gestational age and the incidence of "mature" AF-Lec was not different among the four groups. We conclude that there is no effect of fetal sex, race, or the interaction between them on fetal lung development as measured by AF-Lec. PMID- 2568287 TI - Analysis of 20,291 deliveries in a Turkish institution. AB - Data of 20,291 deliveries are presented; 6.85% (1389) of the patients were 19 years old or under and 6.24% (1276) were 35 or over; 7352 women had one child (primipara) and 1475 had five or more. Cephalic presentation constituted 94.2% of deliveries. The total induction rate was 9%. The incidence of prematurity and post-term gestation was 13 and 7.2%, respectively. Prolonged labor was encountered in 4% of the patients. Anesthesia was employed in 65.8% of all deliveries. Total and primary cesarean section rates were 9.77 and 7%, respectively. The incidence of fetal mortality was 2.55% and congenital anomalies were seen in 1.53% of the infants. The birth weight was under 2499 g in 9.5% of the newborns. Pregnancy complications of different magnitude were encountered in 23.7% of the cases. Uterine rupture and inversion rates were 0.12% and 0.005% respectively. The maternal mortality rate was found to be 64.06 per 100,000 deliveries. PMID- 2568288 TI - Risk factors for fever, endometritis and wound infection after abdominal delivery. AB - Risk factors for postoperative fever, endometritis and wound infection were analyzed in 761 consecutive cesarean sections. Postoperative fever was observed in 12%, endometritis in 4.7% and wound infection in 3% of cases. The relative risk for postoperative fever was increased in cases with postoperative hematoma (relative risk = 16.0), in cases with blood loss over 500 g (relative risk = 1.8) and if the duration of labor exceeded 6 h (relative risk = 1.9). The only significant risk factors for endometritis were amnionitis (relative risk = 8.7), postoperative hematoma (relative risk = 5.0) and age under 24 years (relative risk = 3.0). Wound infections were less frequent in cases with previous cesarean sections (relative risk = 0.15) and after elective cesarean sections (relative risk = 0.22), but duration of operation over 1 h (relative risk = 2.8), induction of labor (relative risk = 3.2) and puerperal endometritis (relative risk = 7.9) increased the risk of wound infection. By elimination of amnionitis and postoperative hematomas the rate of endometritis would have diminished only from 4.7% to 3.8%, a percentage equally unacceptable; diagnostics and prevention should be directed to young patients undergoing caesarean section. Besides technical procedures prevention of endometritis is important for the prevention of wound infection. In hospitals with low postcesarean infectious morbidity antibiotic prophylaxis seems to be unwarranted. PMID- 2568289 TI - Pharmacokinetics of meperidine in pregnancy. AB - The disposition of meperidine was studied in 11 term pregnant humans after the administration of a single 50 mg intravenous dose of meperidine through 48 h post injection. Half-lives of the rapid and terminal elimination phases were calculated as 2.3 and 13.3 h, respectively. These values are much greater than previously reported half-lives which were based on data collected over less than 8 h after injection. An accurate value for t1/2 beta may be particularly important in sequential dosing of analgesic medication. These pharmacokinetic constants calculated on data collected through 48 h in this study may have important clinical correlates. PMID- 2568290 TI - A new method for measurement of surfactant specific apoprotein in amniotic fluid. AB - A simple and rapid method for the quantitative measurement of surfactant specific apoprotein concentration in amniotic fluid was developed using the measurement of immunological reactions by the nephelometric technique. Use of this method made it possible to measure 0.5-8.0 micrograms apoproteins per ml within approximately 70 min. Surfactant specific apoproteins in 54 samples of amniotic fluid were measured using the method. The surfactant specific apoprotein concentration in amniotic fluid increased from 1.03 +/- 0.51 micrograms/ml (mean +/- S.D.) at 26 30 weeks of gestation to 4.45 +/- 2.08 micrograms/ml at 36 weeks of gestation or more. Among premature infants, who were delivered within 24 h afer amniocentesis, surfactant specific apoprotein concentration was less than 1.5 micrograms/ml in three infants with respiratory distress syndrome (RDS), and more than 2.1 micrograms/ml in six without RDS. The results indicated that the quantitative measurement of surfactant specific apoprotein in amniotic fluid is effective in predicting the fetal lung maturity, and that simplicity and rapidity make our method useful for clinical application. PMID- 2568291 TI - Laparoscopic surgery for tubal pregnancies utilizing laser. AB - Nine cases of ectopic pregnancy were treated by laser supported tubotomy and subsequent endoscopic evacuation by a forceps. The location of the ectopic pregnancies were as follows: ampulla (one patient); isthmus (five patients); and fallopian tube (three patients, in two of whom the whole tube was a hematosalpinx). The incision in the tube was made on the antimesenteric side according to the position and size of the ectopic pregnancy using an Nd YAG laser. Tissue destruction of the Nd YAG laser was diminished by using a crystal. After the incision, the conceptional debris was removed by a forceps. In all cases the fallopian tube was left open. Three months after the operation an X-ray was performed. In all nine cases, patent tubes could be documented on the affected and on the contralateral side. In the two patients with hematosalpinx, adhesions of the treated tube were found. PMID- 2568292 TI - A primary amelanotic melanoma of the vagina diagnosed by immunocytochemistry. AB - A case of primary malignant melanoma of the vagina is discussed. The lesion consisted of a nodule in the middle third of the vagina that was histologically suspected of being an unpigmented malignant melanoma. The melanocytic origin of the lesion was confirmed by the pattern of reactivity to a battery of human melanoma associated antigens and to class 1 and 2 histocompatibility antigens. No secondary lesions or alternative primary sites were found. The patient underwent radical hysterectomy with bilateral salpingooophorectomy, total vaginectomy and vulvectomy, radical inguinal, pelvic and paraaortic lymphadenectomy. The pathology report showed the presence of multiple neoplastic foci in the vagina. Although the removed lymph nodes were histologically free of metastases, microscopic foci of neoplastic cells were detected by immunohistochemistry in three lymph nodes. There is no evidence of recurrence at the twelfth postoperative month. Our results show that immunohistochemical techniques may usefully complement diagnostic histopathology in the diagnosis of female genital tract melanoma. PMID- 2568293 TI - An immunohistochemical study of neuroendocrine cells in gynecologic tumors. AB - Various gynecologic tumors with argyrophilia were studied immunohistochemically for chromogranin using two antibodies, antichromogranin and antineuroendocrine. Of seven small cell carcinomas of the cervix, four were immunoreactive with antichromogranin and seven with antineuroendocrine. Argyrophil cells of six cervical adenocarcinomas were all immunoractive with both antibodies. Type I argyrophil cells of 20 endometrial carcinomas were likewise stained positively. However, of the 30 endometrial carcinomas with type II argyrophil cells, 19 showed positive immunoreactivity for chromogranin and 22 for neuroendocrine. Of the ovarian tumors tested, argyrophil cells of 11 mucinous tumors, three carcinoid tumors, and the pancreatic tissue of a malignant mixed germ cell tumor were all chromogranin- and neuroendocrine-immunoreactive. Type I argyrophil cells of five endometrioid carcinomas of the ovary were also immunoreactive with both antibodies. Of the 13 endometrioid carcinomas with type II argyrophil cells, only four showed positive immunoreactivity for chromogranin and only five for neuroendocrine. In conclusion, both antichromogranin and antineuroendocrine detect the specific neuroendocrine markers in close association with argyrophilia in gynecologic tumors, the latter being more sensitive for small cell carcinoma of the cervix, and for type II argyrophil cells in adenocarcinoma of the endometrium and endometrioid carcinoma of the ovary. PMID- 2568294 TI - Comparison of copper T-200 with Lippes loop as a contraceptive device. AB - A comparison has been made between Lippes loop and T Cu 200 intra-uterine contraceptive device with respect to their acceptability and effectiveness in a developing country. A total of 1054 patients were studied; 697 with Lippes loop and 357 with T Cu 200. The ages of the patients ranged between 14 and 44 years, with a peak age of 21-25 years for T Cu 200 and 26-30 for Lippes loop. The event rates were similar but T Cu 200 displayed lower expulsion rate, lower intermenstrual bleeding rate and a lower incidence of pelvic inflammatory disease. The Lippes loop had lower event rate for the other parameters considered, pregnancy and pelvic pain. The continuation rate was similar for both devices. Both devices are useful for women who wish to space out their children but for those who wish to achieve effective contraception for more than 5 years, Lippes loop should be the IUD of choice. PMID- 2568295 TI - Abruptio placentae as complication to acute appendicitis. PMID- 2568296 TI - Repeated contralateral interstitial pregnancy. AB - A case of repeated contralateral interstitial pregnancy is presented in a woman with previous sterility. Biopsies from both uterine corners have later shown salpingitis isthmica nodosa. The etiology, symptoms and treatment are discussed based on the literature. PMID- 2568297 TI - A comparative study of the estrogen receptor ratio in myometrium and uterine leiomyomas. AB - Estrogen receptor (ER) content in myoma and myometrium was determined by radio receptor assay (RRA), and ER staining was performed by immunocytochemical staining, i.e. the peroxidase-antiperoxidase (PAP) method, in uterine samples from eight patients with uterine leiomyoma. ER content in myoma tended to be hither than that in myometrium, but the difference was not significant. The distribution pattern of ER staining was thought to be an important criterion. When the data were compared on the basis of the number of cells in the site where the largest number of positive cells aggregated, the number for myoma was significantly higher than that for myometrium (P less than 0.05). These results suggest that myoma contains more ER positive cells to obtain estrogen effects than myometrium. PMID- 2568298 TI - SSEA-1 is a specific marker for the spinal sensory neuron lineage in the quail embryo and in neural crest cell cultures. AB - Investigation of the early phases of the development of primary sensory neurons has been limited to cells obtained from sensory ganglia. Due to the lack of an early, lineage-specific marker for sensory neuroblasts, it has not been possible to use the neural crest, which gives rise to all spinal and some cranial primary sensory neurons, as a source of precursor cells. In the present study, we show that in neural crest derivatives of the quail embryo, the stage-specific embryonic antigen-1 (SSEA-1) is expressed specifically by developing sensory neuroblasts. The monoclonal antibodies anti-SSEA-1 and AC4 were used to characterize sensory neuron development in vivo and in neural crest cell cultures. In the rat and mouse, both antibodies recognize the same carbohydrate sequence [galactose beta 1-4(fucose alpha 1-3)N-acetylglucosamine] which characterizes SSEA-1. In the quail embryo, this epitope is a marker with several attractive characteristics. Among neural crest derivatives, it is specific for the sensory lineage and is expressed by all detectable sensory neuroblasts at all spinal axial levels. In addition, the carbohydrate sequence appears early and persists throughout development. Expression of SSEA-1 was also studied in neural crest cell cultures, in which two populations of sensory neuroblasts were observed. One population differentiated before or shortly after explanation into culture; these cells did not emigrate from the neural tube. A second population appeared in older cultures. Forming the leading edge of the emigrating neural crest cells, they became SSEA-1+ 3 days after the nonmigrating SSEA-1+ cells. Double staining experiments revealed no obvious differences between the two populations with regard to morphology, neurofilament expression, and neurotransmitter content. PMID- 2568299 TI - Comparison of the effects of placebo, ranitidine, famotidine and nizatidine on intragastric acidity by means of continuous pH recording. AB - The effects of single daily doses of placebo, nizatidine 300 mg, ranitidine 300 mg and famotidine 40 mg, given at 22.00 h, have been compared in 16 patients with healed duodenal ulcers. Each of them underwent the above treatment on four separate occasions. The three H2 receptor antagonists showed a significantly higher acid inhibition than placebo (p less than 0.003) throughout the whole 24 hour period. Famotidine turned out to be more effective than nizatidine (p less than 0.02) in reducing circadian acidity, while there was no difference between ranitidine and nizatidine. The effects of the three H2 blockers on overnight acidity (from 23.00 to 07.00) were similar to each other, whereas both famotidine (p less than 0.003) and ranitidine (p less than 0.02) produced more anacidity than nizatidine during the morning hours (from 07.00 to 12.00). The time period elapsed in consecutive minutes above 5.0 pH units during drug-related events was significantly longer with both famotidine (p less than 0.01) and ranitidine (p less than 0.01) compared to nizatidine. Therefore in the recommended dosages for clinical use the acid suppression of nizatidine was significantly shorter-lasting than that of both ranitidine and famotidine. The major change is represented by the lack of carryover effect of nizatidine on morning acidity. PMID- 2568300 TI - Pharmacological effect of somatostatin on bile secretion in man. AB - Somatostatin (SST) has been shown to induce cholestasis in the dog and in the rat. In man, it is still unknown whether SST modifies bile formation. The present study was undertaken to examine the influence of SST on bile secretion in man. Two volunteers who had a total external biliary fistula received 1-hour SST infusions (3.5 micrograms/kg/h). Bile flow, bile acid, phospholipid and cholesterol biliary outputs were measured before, during and 1 h after the infusion. The SST infusion was associated with a pronounced decrease in bile flow and in bile acid secretion and with an increase in bile cholesterol saturation. These findings suggest that SST has cholestatic properties in man as in other species. This may provide a rational explanation for the formation of gallstones and for the steatorrhea observed in patients with somatostatinomas or during therapeutic SST administration. PMID- 2568301 TI - The guanylate cyclase/receptor family of proteins. AB - Guanylate cyclase, which catalyzes the formation of cGMP from GTP, exists in both the soluble and particulate fractions of cells. At least two different cellular compartments for the particulate enzyme exist: the plasma membrane and cytoskeleton. The enzyme form found in the soluble fraction is a heterodimer that can be regulated by free radicals and nitrovasodilators, whereas the membrane form exists as a single-chain polypeptide that can be regulated by various peptides. These peptides include resact and speract obtained from eggs and atrial natriuretic peptides (ANP). The species of guanylate cyclase present in cytoskeletal fractions resists solubilization with non-ionic detergents; its structural properties are not yet known. cDNAs encoding the membrane form of guanylate cyclase have been isolated from different tissues and species, and in all cases the DNA sequences predict a protein containing a single transmembrane domain. The carboxyl (intracellular) domain is highly conserved from sea urchins through mammals, whereas the extracellular domain (amino terminus) varies considerably. The predicted amino acid sequences demonstrate that the membrane form of guanylate cyclase is a member of a diverse and complex family of proteins that includes a low molecular weight ANP receptor, protein kinases, and the cytoplasmic form of guanylate cyclase. cDNA encoding a membrane form of the enzyme from mammalian tissues has been expressed in cultured cells, and the expressed guanylate cyclase specifically binds ANP and is activated by ANP. The membrane form of guanylate cyclase, then, serves as a cell surface receptor, representing the first recognized protein to directly catalyze formation of a low molecular weight second messenger in response to ligand binding. PMID- 2568302 TI - Location cloning strategy for characterizing genetic defects in Huntington's disease and Alzheimer's disease. AB - The recognition that DNA polymorphisms are widespread in the human genome and can be used as high quality genetic markers has introduced a new strategy for approaching inherited disorders for which no protein defect has been identified. Genetic linkage analysis can establish the chromosomal position of the genetic defect, providing a potential opportunity for isolating the disease gene and characterizing its product in the absence of any knowledge of its biochemical function. The first step in this location cloning approach has been successful in mapping the Huntington's disease gene to chromosome 4, and has implicated chromosome 21 as the site of a defect in familial Alzheimer's disease. An intensive effort is under way to narrow the region containing the disease gene and identify the defect in each of these disorders. This review will present the success that has been achieved and the problems that remain and will assess the current status of the location cloning strategy with regard to Huntington's disease and familial Alzheimer's disease. PMID- 2568303 TI - Effect of a new H2-receptor antagonist (roxatidine acetate) on the acid secretion by isolated canine parietal cells. PMID- 2568304 TI - Effect of folate supplementation on the incidence of dysplasia and cancer in chronic ulcerative colitis. A case-control study. AB - Folate deficiency has been associated with dysplasia in human cancer models. Patients with ulcerative colitis commonly have decreased folate levels, which are partially due to sulfasalazine, a competitive inhibitor of folate absorption. To study the effect of folate supplementation on the risk of dysplasia or cancer (neoplasia) in ulcerative colitis, records from 99 patients with pancolitis for greater than 7 yr and enrolled in a surveillance program were reviewed. Thirty five patients with neoplasia were compared with 64 patients in whom dysplasia was never found to determine the effect of folate supplementation on the rate of development of neoplasia using case-control methodology. At the time of the index colonoscopy, patients with neoplasia were older (43 +/- 11 vs. 39 +/- 12 yr) and had disease of longer duration (20 +/- 8 vs. 15 +/- 7 yr, p less than 0.05). Folate supplementation was associated with a 62% lower incidence of neoplasia compared with individuals not receiving supplementation (odds ratio, 0.38; 95% confidence interval, 0.12-1.20). There was no appreciable change in this effect when models were fit to adjust for sulfasalazine dose, duration of disease, age at symptom onset, prednisone dose, sulfa allergy, sex, race, or family history of colon cancer. The statistical power of the association between folate supplementation and neoplasia was 72%. Correction of risk factors before the development of neoplasia may prevent this serious complication. Pending a larger case-control study, folate supplementation during sulfasalazine administration is recommended to possibly prevent the complication of dysplasia or cancer in ulcerative colitis. PMID- 2568305 TI - Pharmacological activity profiles of dopamine D-1 and D-2 reception agonists and antagonists on striatal neuronal activity and the response to dexamphetamine in freely moving rats. AB - 1. The effects of dopamine D-1 and D-2 receptor agonists and antagonists were investigated by recording extracellular striatal action potentials in freely moving rats. Dopamine receptor antagonist effects were also evaluated on dexamphetamine-induced excitation of striatal neurons. 2. Striatal neurons responded to SKF 38393, a D-1 agonist, with dose-dependent reductions in activity. At a 2.0 mg/kg dose neuronal activity decreased to 50% of control values. 3. The D-1 antagonist, SCH 23390, at a dose of 4.0 mg/kg decreased striatal neuronal activity by more than 50% and also effectively blocked the effects of 2.5 mg/kg dexamphetamine. 4. LY 171555, a D-2 agonist, at 1.0 or 2.5 mg/kg, did not significantly increase striatal neuronal activity. Although behavioral activation was noted, the neuronal response at the high dose was biphasic with inhibition predominant. 5. The D-2 antagonists haloperidol and sulpiride decreased striatal neuronal activity in a dose-dependent manner and also effectively antagonized the effects of dexamphetamine. The D-2 antagonist, RO 22-1319, at a dose of 2.0 mg/kg completely antagonized increases in striatal neuronal activity after dexamphetamine. 6. These findings suggest that dexamphetamine-induced increases in striatal neuronal activity are due to either stimulation of both D-1 and D-2 receptors, or alternatively, a third dopamine receptor subtype sensitive to both D-1 and D-2 antagonists but not agonists. Furthermore, the concept of specific D-1 and D-2 receptor agonists may require revision as neither SKF 38393 or LY 171555 increased striatal neuronal activity. PMID- 2568307 TI - Excitatory action of the native neuropeptide antho-rfamide on muscles in the pennatulid Renilla kollikeri. AB - 1. Antho-RFamide (pGlu-Gly-Arg-Phe-amide), a neuropeptide recently isolated from the sea pansy Renilla kollikeri induced sustained (tonic) contractions in the rachis and peduncle of the colony, and in the individual autozooid polyps. 2. The threshold concentration for this effect was 5 nM in summer colonies and 1 microM in autumn or winter colonies. 3. The peptide-induced tonic contractions were unaffected in sodium-free sea water. There was a 30% reduction of the contraction amplitude in sea water lacking calcium. 4. Peptides related to Antho-RFamide and other peptides were also examined for activity on rachidial muscles. Only peptides containing the carboxyterminal sequence Arg-Phe-amide were active. 5. It is concluded that Antho-RFamide acts on Renilla muscles via a specific receptor and that it is a candidate for neurotransmitter or modulator in this pennatulid. PMID- 2568306 TI - Lipolytic response of bovine adipose tissue to alpha and beta adrenergic agents 30 days pre- and 120 days postpartum. AB - 1. The beta adrenergic agonists isoproterenol and epinephrine stimulated in vitro lipolysis in adipose tissue removed from heifers 30 days pre- and 120 days postpartum. 2. Propranolol, a beta antagonist, blocked isoproterenol stimulated lipolysis pre- and postpartum. 3. Epinephrine co-incubated with propranolol resulted in a suppression of lipolysis similar to that produced by clonidine, an alpha agonist, both pre- and postpartum. 4. Bovine adipose tissue lipolysis was more responsive to isoproterenol and isoproterenol + propranolol at 120 days of lactation than 30 days prepartum. 5. Adipose tissue sensitivity to clonidine, epinephrine and epinephrine + propranolol did not differ 30 days pre- and 120 days postpartum. PMID- 2568308 TI - Chromosomal mapping of genes encoding mannose-sensitive (type I) and mannose resistant F8 (P) fimbriae of Escherichia coli O18:K5:H5. AB - DNA hybridization experiments demonstrated that the gene clusters encoding the F8 fimbriae (fei) as well as the type I fimbriae (pil) exist in a single copy on the chromosome of E. coli O18:K5 strain 2980. In conjugation experiments with appropriate donors, the chromosomal site of these gene clusters was determined. The pil genes were mapped close to the gene clusters thr and leu controlling the biosynthesis of threonine and leucine, respectively. The fei genes were found to be located close to the galactose operon (gal) between the position 17 and 21 of the E. coli chromosomal linkage map. PMID- 2568309 TI - Extrapyramidal reactions. Neuropsychiatric mimics in patients with AIDS. AB - The use of neuroleptic medication as antiemetics, or in the treatment of neuropsychiatric disorders in patients with AIDS, may be associated with extrapyramidal side effects and lead to difficulty with diagnosis and management. Two cases are presented that describe severe extrapyramidal syndromes occurring in two patients with AIDS, one treated with prochlorperazine and the other with prochlorperazine and metoclopropramide. It is possible that the neuropathologic lesions found in patients with the AIDS dementia complex may pre-dispose to extrapyramidal side effects of neuroleptic medication. The differential diagnosis and treatment of delirium, dementia, depression, and extrapyramidal reactions in patients with AIDS is discussed. PMID- 2568310 TI - Variable evolutionary stability of Y chromosomal repeated sequences in the genus Mus. AB - The study reported here is an examination of the organization and evolution of three Y chromosomal repeated sequences, designated pBC10-0.6, pBC15-1.1, and pBA33-1.8, in five closely related species of the genus Mus. The species distributions of major restriction fragment length polymorphisms produced with a panel of restriction enzymes is used to develop the phylogenetic relationships between the five species studied. However, the apparent degree of relatedness among these species varied a great deal with each of the three probes and was also highly dependent on the particular restriction enzyme used. The usefulness for phylogenetic studies of closely associated sequences varying in evolutionary stability is discussed. PMID- 2568311 TI - Complementary homeo protein gradients in developing limb buds. AB - A new human homeo box-containing gene designated Hox-5.2 was cloned and mapped to human chromosome 2. This homeo box is related in sequence to Abdominal-B, a Drosophila homeotic gene that specifies identity of posterior segments. An antibody probe was made using a human Hox-5.2 fusion protein and was found to stain posterior regions of mouse, chicken, and Xenopus embryos. Unexpectedly, when the distribution of Hox-5.2 antigen was compared with that of X1Hbox 1 antigen, a non-overlapping and mutually exclusive pattern was detected (e.g., in developing limb buds, intestine, and somites). Regions expressing Hox-5.2 do not express X1Hbox 1 protein, and vice versa. Hox-5.2 antigen is detected strongly in developing fore- and hindlimb buds, where it forms a gradient of nuclear protein throughout most of the mesenchyme. This gradient is maximal in distal and posterior regions. Hox-5.2 expression is activated in Xenopus limb regeneration blastemas, as expected for any gene involved in pattern formation. As described previously, a gradient of X1Hbox 1 protein can be detected in the forelimb. The latter gradient has the opposite polarity to that of Hox-5.2. i.e., maximal in anterior and proximal mesoderm. These two opposing gradients (and possibly others) could be involved in determining positional values in developing limb buds. PMID- 2568312 TI - The DNA-binding homeo domain of the Oct-2 protein. PMID- 2568314 TI - Expression of v-src induces a myeloproliferative disease in bone-marrow reconstituted mice. AB - A recombinant retrovirus, N-TK-src, was used to introduce the v-src oncogene into mouse hematopoietic cells. This vector efficiently expresses both the neo and v src genes in different hematopoietic lineages in culture as well as in mice reconstituted with infected bone marrow cells. Expression of v-src had no dramatic effect on the proliferative and differentiative capacity of hematopoietic precursors when assayed in methyl cellulose cultures. However, in mice reconstituted with N-TK-src-infected bone marrow cells, expression of v-src leads to the rapid development of a severe myeloproliferative disease, characterized by splenomegaly, anemia, and a shift of hematopoiesis from the bone marrow to the spleen. PMID- 2568313 TI - Poly(A) elongation during Xenopus oocyte maturation is required for translational recruitment and is mediated by a short sequence element. AB - Xenopus oocytes contain several mRNAs that are mobilized into polysomes only at the completion of meiosis (maturation) or at specific times following fertilization. To investigate the mechanisms that control translation during early development, we have focused on an mRNA, termed G10, that is recruited for translation during oocyte maturation. Coincident with its translation, the poly(A) tail of this message is elongated from approximately 90 to 200 adenylate residues. To identify the cis sequence that is required for this cytoplasmic adenylation and recruitment, we have synthesized wild-type and deletion mutant G10 mRNAs with SP6 polymerase. When injected into oocytes that subsequently were induced to mature with progesterone, wild-type G10 mRNA, but not mutant transcripts lacking a 50-base sequence in the 3'-untranslated region, was polyadenylated and recruited for translation. The 50-base sequence was sufficient to confer polyadenylation and translation when fused to globin mRNA, which does not normally undergo these processes during oocyte maturation. Further mutational analysis of this region revealed that a U-rich sequence 5' to the AAUAAA hexanucleotide nuclear polyadenylation signal, as well as the hexanucleotide itself, were both required for polyadenylation and translation. The 50-base cis element directs polyadenylation, but not translation per se, as a transcript that terminates with 3'-deoxyadenosine (cordycepin) is not recruited for translation. The available data suggest that the dynamic process of polyadenylation, and not the length of the poly(A) tail, is required for translational recruitment during oocyte maturation. PMID- 2568315 TI - Human gamma-glutamyl transpeptidase cDNA: comparison of hepatoma and kidney mRNA in the human and rat. AB - gamma-Glutamyl transpeptidase (GGT) is a glutathione-metabolizing enzyme that has been extensively studied in relation to hepatocarcinogenesis. Using a cDNA for rat kidney GGT as a probe, we have isolated a full-length cDNA for human GGT from a hepatoma cell-line library. Nucleotide sequence analysis of the clone revealed a 2326-bp insert that includes a 5'-untranslated region of 487 nucleotides (nt), an open reading frame (ORF) of 1707 nt, and a 3'-untranslated region of 132 nt. The ORF encodes a protein with an amino acid sequence that is highly similar to that of the rat GGT precursor peptide, with an overall identity of 79%. The cDNA clone was used to probe Northern blots of hepatoma and kidney RNA from both human and rat. In both species, the GGT mRNA is longer in hepatoma than in kidney. In addition, the human mRNAs were longer than their counterparts in the rat. None of three human hepatocellular carcinomas examined showed a marked elevation in GGT mRNA levels relative to surrounding liver tissue. PMID- 2568316 TI - Expression of human erythropoietin cDNA in human lymphoblastoid Namalwa cells: the inconsistency of a stable expression level with transient expression efficiency. AB - Recombinant plasmids for the expression of human erythropoietin (EPO) cDNA in Namalwa cells were constructed. From the results of the EPO expression efficiency in transiently transfected cells, it was found that the simian virus 40 (SV40) early promoter directs EPO synthesis more efficiently in Namalwa cells than does the long terminal repeat promoter of Rous sarcoma virus and that the 3'-noncoding sequence including splice junction and polyadenylation site derived from the rabbit beta-globin gene are more effective than those of the SV40 early gene. However, in stable transformants, no simple relationship was found between the expression level of EPO cDNA and the structure of the introduced expression vectors. PMID- 2568318 TI - Interdisciplinary issues in mental health and aging. AB - An Interdisciplinary Conference on Mental Health and Aging held in Washington, D.C. in May 1986 restimulated the interdisciplinary cooperation generated by the 1980 White House Mini-Conference on the Mental Health of Older Adults and provided a continuing basis for interdisciplinary promotion of coordinated geriatric mental health services. The Conference reflected a wide-spread recognition of the need for interdisciplinary cooperation and the coordination of services for the elderly. The overall organization of the meeting is provided here and articles based on the presentations given by representatives of the four core mental health disciplines follow. PMID- 2568317 TI - The chicken homeo box genes CHox1 and CHox3: cloning, sequencing and expression during embryogenesis. AB - Several Drosophila genes involved in the control of segmentation and segment identity share a 183-bp conserved sequence termed homeo box. Homeo box sequences have been detected and cloned from the genomes of insects like Drosophila to vertebrates such as mouse and man. Two chicken homeo box genes CHox1 and CHox3, are described. Cloning of the CHox1 and CHox3 homeo boxes was performed using Drosophila and murine homeo box sequences as probes under low-stringency conditions. Analysis of both chicken homeo box sequences revealed them to be homeo boxes that have diverged from the Antennapedia class with homologies to homeo boxes of other organisms in the range of 75-42% at the nucleotide level and 69-41% at the protein level. Analysis of CHox3 expression during early embryo development showed that the gene codes for five transcripts 1.3, 1.9, 2.6, 5.6 and 7.9 kb in size. Three of the transcripts (1.3, 1.9 and 5.6 kb) are also recognized by a flanking non-homeo box containing probe. The levels of the different transcripts changed during the first five days of development. The most abundant transcripts (1.3 and 1.9 kb) are already present at the time the egg is laid. Their transcription peaks at day 1 of incubation and then decreases. The CHox1 transcripts are present at very low levels between days 2.5 and 4 of development. These two chicken genes represent bona fide Hox genes in a branch of vertebrates that evolved parallel to mammals. PMID- 2568319 TI - Conference on interdisciplinary issues in mental health and aging: workshop issues and recommendations. AB - The Conference of Interdisciplinary Issues in Mental Health and Aging included four workshops which focused on issues pertaining to the well elderly, the short term impaired elderly, the chronic mentally ill elderly, and the organically impaired elderly. The Conference was convened to review and assess the impact of policy and program changes in the field of mental health and aging since the 1981 White House Conference on Aging and to reaffirm and strengthen interdisciplinary cooperation in addressing the mental health needs of older persons. This article presents the issues discussed in the workshops and recommendations regarding services, research, financing, and training and education which resulted from these sessions. PMID- 2568320 TI - [Reorganization of the neuromediatory function of the brain and autonomic regulation at rest during adaptation to chronic hypokinesia]. PMID- 2568324 TI - International recommendations (as of March, 1988) on the application of methods involving DNA-polymorphisms in forensic haematology. PMID- 2568321 TI - [Urinary excretion of hydroxyproline after exposure to quartz dust and glutamic acid]. PMID- 2568322 TI - [Debatable problems of intravascular dilatation of the brachiocephalic artery]. AB - Roentgen--guided endovascular dilatation (RED) was conducted in 23 patients with atherosclerotic stenoses of branches of the arch of the aorta, in 20 with stenoses of the subclavian artery, and in 3 with stenoses of the brachiocephalic trunk. Before dilatation the patients had a systolic pressure gradient of 25 to 55 mm Hg in the region of the stenosis and constriction of the arterial lumen by 40 to 80%. Systolic pressure gradient disappeared completely in 17 cases and persisted at a level of 10 to 40 mm Hg in 7 cases. During RED of the left subclavian artery a complication, acute thrombosis of the distal part of the artery, occurred in one case. Indications for dilatation of stenosed subclavian arteries were elaborated on the basis of the accumulated experience. PMID- 2568323 TI - Spectrum of HIV infection and AIDS in a cohort of Italian hemophiliacs. AB - A group of 173 subjects affected by congenital clotting factor deficiencies was evaluated with regard to the impact of HIV infection. On the whole, 78 patients (45%) were found to be HIV Ab-positive. As of March, 1988, of the seropositive patients, 63 (80.8%) had an asymptomatic HIV infection (Group II/CDC), three (3.8%) had a persistent generalized lymphadenopathy (Group III/CDC). The 12 (15.4%) remaining patients could be classified in Group IV of the CDC classification due to their symptoms and signs; in particular, 10 came under surveillance case definition for AIDS. Assay for the HIV antigen was positive in 14 (17.9%) seropositive hemophiliacs. With regard to the immunological features, our data clearly show that a sharp decline in the number of CD4+ cells was associated with symptomatic forms of the disease. An evaluation of the time elapsed from seroconversion to the appearance of the symptomatic clinical condition showed an average incubation of 37 months. PMID- 2568325 TI - Acute effects with reference to thrombogenicity and liver toxicity after injection of different coagulation factor concentrates. AB - In order to assess thrombogenicity and liver toxicity of different coagulation factor concentrates, antithrombin III, soluble fibrin, alanine aminotransferase (ALAT) and gamma-glutamyl transpeptidase (GT) were measured in samples taken before, 30 min and 24 h after the infusion. Seventy-six studies were performed in 55 patients with haemophilia A (37), B (11) or von Willebrand's disease, type III (7). A sharp rise of soluble fibrin was observed in 2 patients with haemophilia B, indicating that modern factor IX concentrates may still be thrombogenic. A slight increase was also seen after infusion of factor VIII-von Willebrand factor concentrates of low purity. Antithrombin III, ALAT and GT did not change significantly after any of the factor concentrates. The alterations in those parameters did not correlate with the impairment of liver function. PMID- 2568326 TI - [Neuropeptide Y (NPY): functions and biosynthesis as a peptidergic neurotransmitter and the regulation of neuron-specific expression of NPY gene]. AB - Neuropeptide Y (NPY) is widely distributed in the central and sympathetic nervous systems and has a variety of central actions including regulation of blood pressure and peripheral actions; e.g., continuous vasoconstriction and inhibition of catecholamine release. The NPY receptor can be divided into 2 subclasses (Y1, Y2), and these subclasses are coupled to GTP binding proteins (Gi, Go, Gp ......). Recently, human and rat prepro-NPY mRNA and NPY gene structures have been determined by cDNA and genomic cloning and sequencing. The strong evolutionary conservation of these structures suggested that NPY is an essential peptidergic neurotransmitter. Little is known about the biosynthesis, processing, degradation of NPY and NPY gene expression. We showed that NPY gene expression and NPY biosynthesis are regulated by neural activity, hormone, and intracellular second messengers via neurotransmitter receptors. The change of NPY gene expression by these neural factors is considered to be a good model for a synaptic plasticity, because these changes cause the changes of synaptic transmission. Furthermore, because NPY is expressed in sympathetic neurons and its gene expression increased markedly on the differentiation of adrenergic cells, this study about NPY gene expression could provide good clues for elucidating the differentiation of sympathetic neurons. PMID- 2568327 TI - [Treatment of acute stomach ulcer with H2 receptor antagonists. A direct therapy comparison with 300 mg nizatidine nightly and 2 times 150 mg nizatidine with twice daily 150 mg ranitidine]. AB - In this endoscopically controlled, double-blind study involving 242 patients with acute benign gastric ulcer, it was shown that the selective inhibition of nocturnal gastric acid secretion with 300 mg nizatidine administered on retiring represents an effective and reliable form of therapy. After four weeks of treatment, 90% of the patients receiving 300 mg nizatidine, no longer experienced nocturnal pain, in comparison with 83% receiving 2 X 150 mg nizatidine daily (n.s.). The total healing rates after four weeks were 60% in the patients receiving 300 mg nizatidine, 60% in those on 2 X 150 mg nizatidine daily, and 58% in those receiving 2 X 150 mg ranitidine daily. After eight weeks, the respective figures rose to 85%, 84% and 84%. Clinically relevant side effects were observed in none of the three groups. With a dose on retiring of 300 mg nizatidine, it is possible to accelerate the healing of a benign gastric ulcer, simply by the nocturnal suppression of gastric acid production, and, during the day, preserving physiological gastric function, thus avoiding the possible risks of protracted hypochlorhydria. PMID- 2568328 TI - Norrie disease: linkage analysis using a 4.2-kb RFLP detected by a human ornithine aminotransferase cDNA probe. AB - Previous study has shown that the usual DNA marker for Norrie disease, the L1.28 probe which identifies the DXS7 locus, can recombine with the disease locus. In this study, we used a human ornithine aminotransferase (OAT) cDNA which detects OAT-related DNA sequences mapped to the same region on the X chromosome as that of the L1.28 probe to investigate the family with Norrie disease who exhibited the recombinational event. When genomic DNA from this family was digested with the PvuII restriction endonuclease, we found a restriction fragment length polymorphism (RFLP) of 4.2 kb in size. This fragment was absent in the affected males and cosegregated with the disease locus; we calculated a lod score of 0.602, at theta = 0.00. No deletion could be detected by chromosomal analysis or on Southern blots with other enzymes. These results suggest that one of the OAT related sequences on the X chromosome may be in close proximity to the Norrie disease locus and represent the first report which indicates that the OAT cDNA may be useful for the identification of carrier status and/or prenatal diagnosis. PMID- 2568330 TI - A genetic linkage map of 17 markers on human chromosome 21. AB - We have constructed a genetic linkage map of 17 markers on the long arm of human chromosome 21, including six genes and two anonymous loci with a variable number of tandem repeats. The estimated length of the map is 103 cM in males and 140 cM in females, assuming Kosambi interference. Recombination in females was approximately twice that in males between proximal markers. However, over half of the recombination events in either sex occur distally, in 21q22.3, although this region accounts for only about 15% of the physical length of chromosome 21. PMID- 2568329 TI - Linkage analysis of the murine mos proto-oncogene on chromosome 4. AB - A linkage analysis of the murine Mos gene, which codes for the c-mos proto oncogene, was performed in 88 backcross progeny of an interspecies cross of laboratory mice and Mus spretus. Linkage was tested for four different genes on mouse chromosome 4: Aco-1, Mup-1, b, and Ifb. The gene order (from centromere) with intervening percentage recombination is Mos-15.9 (+/- 3.9)-Aco-1-5.6 (+/- 2.4)-Mup-1-3.4 (+/- 1.9)-b-5.6 (+/- 2.4)-Ifb. These results confirm the previous assignment of Mos to chromosome 4 on the basis of segregation in somatic cell hybrids (D. Swan et al., 1982, J. Virol. 44: 752-754) and show furthermore that Mos and the Ifa/Ifb clusters are not tightly linked as a group of intronless genes, but are separated by a map distance of 30.6 +/- 4.9 recombination units. The linkage data obtained in the present study place Mos in a region compatible with the physical map (D. W. Threadgill and J. E. Womack, 1988, Genomics 3: 82 86). PMID- 2568331 TI - Identification of a 220-kb insertion into the Duchenne gene in a family with an atypical course of muscular dystrophy. AB - Most known mutations in the gene region responsible for Duchenne or Becker muscular dystrophy are deletions of varying extent. Here we describe a 220-kb insertion within the DMD/BMD gene that cosegregates with a somewhat atypical course of muscular dystrophy in a pedigree. The insertion is demonstrated by field-inversion gel electrophoresis as an enlarged SfiI fragment hybridizing to probe J-Bir, while neighboring SfiI fragments (detected by probes PERT 87 and J 66) are unchanged. Hybridization with DMD c-DNA probes did not reveal alterations in coding sequences. In this pedigree, the altered SfiI fragments provide convenient markers for carrier identification. PMID- 2568332 TI - Linkage analysis of a large Latin-American family with X-linked retinitis pigmentosa and metallic sheen in the heterozygote carrier. AB - An extended linkage analysis was performed on the large Latin-American kindred with X-linked retinitis pigmentosa (XLRP) and metallic sheen in the heterozygous carrier studied and reported previously by R.L. Nussbaum et al. (1985, Hum. Genet. 70:45-50) and on a smaller family with the same XLRP variant. In these kindreds the XLRP locus shows close linkage with Xp21 marker loci OTC and DXS206. The results of this linkage analysis agree with the observations made by Nussbaum et al. (1985) that an XLRP locus is distal to DXS7. PMID- 2568333 TI - The role of beta blocker in the post operative low output state following adequate valvotomy in cases of pure valvar pulmonary stenosis (clinical experience). AB - The role of infundibular resection in cases of pure valvar pulmonary stenosis is much debated. Some advocate aggressive approach especially in cases of supra systemic right ventricular pressures. Some consider it unnecessary, and feel that it is secondary hypertrophy which will regress in due course following a successful valvotomy. But all agree that the dynamic right ventricular outflow obstruction plays a very crucial role in the immediate post operative period which can cause significant morbidity and mortality. We advocate the use of beta blocker to relieve the infundibular spasm in the immediate post operative period, thereby relieving the dynamic outflow obstruction. We herewith present two cases to exemplify this. PMID- 2568336 TI - Localization of heat shock protein 70 genes inside the rat major histocompatibility complex close to class III genes. PMID- 2568334 TI - C4A gene deletion and HLA associations in black Americans with systemic lupus erythematosus. AB - In North America and European Caucasoids with systemic lupus erythematosus (SLE) there is an increased frequency of a C4A, CYP21A gene deletion, largely associated with the HLA-B8,DR3,C4A*QO extended haplotype. There have been no consistent HLA associations reported for SLE in blacks, although an increased frequency of serologically determined C4A null alleles has been reported in two studies. We studied 79 black American SLE patients and 68 black controls by restriction fragment length polymorphism analysis to determine if a C4A gene deletion was a genetic risk factor for SLE. Moreover, the nature of the deletion and any HLA phenotypic associations were sought. Nineteen of 79 (24%) patients compared to 5 of 68 (7.4%) controls had a phenotypic C4A,CYP21A gene deletion (P = .005; RR = 4). A homozygous deletion in four patients gave a genotypic frequency of 23/158 (14.5%) SLE patients vs 5/136 (3.7%) controls (P = .001; RR = 4.5). The deletion was associated with HLA-DR2 (P = .03) and HLA-DR3 (P = .03). Moreover, all subjects with the deletion had HLA-DR2 or DR3 (P = 7.7 x 10(-6). HLA-B44 was also associated with the deletion (P = .02), and eight of the nine HLA-B44 positives also carried HLA-DR2. HLA-B8 approached significance (P = .08) and was always accompanied by HLA-DR3. Finally, this black population demonstrated a unique C4B gene size polymorphism with 80% C4B "short" as compared to the 40% C4B "short" frequency reported in whites. We conclude that a large C4A,CYP21A gene deletion, particularly associated with the HLA-B44, -DR2, and DR3 alleles, is the strongest genetic risk factor thus far identified for SLE susceptibility in black Americans. Furthermore, the unique preponderance of the C4B "short" gene form may be a factor in the actual formation of the deletion. PMID- 2568335 TI - A mouse chromosome 17 gene encodes a testes-specific transcript with unusual properties. AB - We have characterized a novel mouse gene (D17Si11) on chromosome 17 that expresses a major transcript observed uniquely in the testes. The D17Si11 locus has been mapped to the central region of chromosome 17 between H-2 and C3. Sequence analysis demonstrates several unusual features of this locus and its transcript: first is the presence of complementary sets of alternating purine and pyrimidine residues within the 3' region of the transcript that could form double stranded, hairpin-like secondary structures with properties similar to that of Z DNA; second is the existence of a hypothetical, long open reading frame in the nucleotide strand that is complementary to the testes transcripts. This complementary strand open reading frame is three times the size of the longest potential open reading frame present in the transcript itself. Although a function for D17Si11 has yet to be determined, the gene is relatively nonpolymorphic in mice and appears conserved in mammals. PMID- 2568337 TI - Lack of close linkage between human coagulation factor F13A and HLA class I molecules. PMID- 2568338 TI - Definition of HLA class I-associated DNA polymorphisms using an HLA-B locus derived cDNA probe. PMID- 2568339 TI - Analysis of HLA-B35 variants and B35 haplotypes by isoelectric focusing and Southern blot analysis. PMID- 2568340 TI - Sulphasalazine desensitisation. PMID- 2568341 TI - Astroglial cells in vitro are heterogeneous with respect to expression of the alpha 1-adrenergic receptor. AB - A wide variety of approaches have been used to examine the expression of neuroligand receptors by cultured astroglial cells. The results of such studies make it clear that these cells exhibit most, if not all, of the different receptors known to be associated with central neurons. However, it has been more difficult to determine if there are multiple populations of astroglia that can be distinguished on the basis of their complement of neuroligand receptors. To address this question, we established a radioligand binding assay that enabled us to visualize alpha 1-adrenergic receptors (alpha 1-ARs) on immunocytochemically defined neural cells. Saturation, time course, and competition binding experiments determined that 125I-HEAT could be used to identify alpha 1-AR binding sites on immunocytochemically defined astroglial cells. Our results indicate that approximately 66% of cortical polygonal astroglia express detectable numbers of alpha 1-ARs. 3H-thymidine labeling experiments indicate that both dividing and nondividing astroglia exhibit alpha 1-ARs. These results greatly contrast with studies on beta-adrenergic receptor expression (beta-AR), in which 100% of polygonal astroglia express beta-ARs. Process-bearing astroglia also exhibit alpha 1-ARs, which is in marked contrast to the lack of beta-AR expression by these cells. The results presented here suggest that astroglia, like neurons, differ in the receptors they express and therefore may be able to selectively respond to extracellular stimulation. PMID- 2568342 TI - Activation of polyphosphoinositide metabolism as a signal-transducing system coupled to excitatory amino acid receptors in astroglial cells. AB - Excitatory amino acids (EAA) are known to induce an increase in the breakdown of polyphosphoinositides (PI) in brain slices and in dispersed cultures of neurons. We have now used astroglia cultured from newborn rat cerebra to demonstrate that glutamate provokes, in [3H]inositol-labeled cells, an accumulation of inositol phosphates in a time- and concentration-dependent manner. The ED50 value for glutamate was 40 microM. Quisqualate, ibotenate, and kainate were also active, with their relative potencies in the order of quisqualate greater than ibotenate much greater than kainate. No effect was detected with N-methyl-D-aspartate and quinolinic acid in the absence of Mg2+. The nonselective glutamate receptor antagonist gamma-D-glutamylglycine fully inhibited glutamate agonist-induced PI breakdown. A brief pretreatment of the astroglial cells with phorbol esters negated these effects of EAA receptor agonists, suggesting a feedback role for protein kinase C in phospholipase C action. Glutamate also elevated cytosolic free Ca2+ in Fura-2-loaded astroglial cells, as assessed by digital fluorescence imaging microscopy. Since a close metabolic partnership is known to exist between neurons and glia, these findings may have important functional consequences for neural cells in vivo. PMID- 2568344 TI - Adherence targets of Vibrio parahaemolyticus in human small intestines. AB - Formalin-fixed human small intestinal mucosa with mucus coating, villi, and lymphoid follicle epithelium at the mucosal surface was used to test the adherence sites of clinically isolated (Kanagawa phenomenon-positive) strains of Vibrio parahaemolyticus. V. parahaemolyticus strains grown on CFA agar (supplemented with 3% NaCl) for ca. 3 h at 37 degrees C possessed various levels of cell-associated hemagglutinins (HAs) which were detected with human or guinea pig erythrocytes. The observed adherence abilities of V. parahaemolyticus strains to human small intestinal mucosa correlated roughly with the HA levels of the strains. Under the test conditions, ileal lymphoid follicle epithelium (especially M cells) provided the best adherence target for V. parahaemolyticus. Adherence to villus absorptive cells or to mucus coating was observed at lower levels. In addition, all 3-h-grown V. parahaemolyticus strains tested produced high levels of HAs as detected with rabbit erythrocytes. The strains were all strikingly motile. In contrast, V. parahaemolyticus strains grown on CFA agar (supplemented with 3% NaCl) for ca. 20 h at 37 degrees C had much lower levels of HAs, adherence abilities, and motility. In contrast to the above observations, piliation of V. parahaemolyticus was more extensive at ca. 20 h of incubation at 37 degrees C than at ca. 3 h of incubation at 37 degrees C. The remarkable ability of V. parahaemolyticus to adhere to lymphoid follicle epithelium was also confirmed by using rabbit small intestinal mucosa. PMID- 2568343 TI - Contribution of autolysin to virulence of Streptococcus pneumoniae. AB - Insertion-duplication mutagenesis was used to construct an autolysin-negative derivative of Streptococcus pneumoniae. This derivative was obtained by first transforming the nonencapsulated strain Rx1 with a derivative of the vector pVA891 carrying a 375-base-pair TaqI DNA fragment from the middle of the autolysin structural gene. DNA was extracted from the resultant erythromycin resistant, autolysin-negative rough pneumococcus and used to transform S. pneumoniae D39, a virulent type 2 strain. Several erythromycin-resistant transformants were obtained from two independent experiments, and none of these transformants produced autolysin. Southern blot analysis confirmed that the autolysin gene in these transformants had been interrupted by the plasmid-derived sequences. The autolysin-negative mutants showed markedly reduced virulence for mice compared with that of strain D39; intranasal and intraperitoneal 50% lethal doses were increased 10(2)- and 10(5)-fold, respectively. Autolysin production was reinstated in one of the mutants by back-transformation with the cloned autolysin gene, with the concomitant loss of erythromycin resistance; the virulence of this isolate for mice was indistinguishable from that of D39. The importance of autolysin in pathogenesis was confirmed by immunization-challenge studies. Mice immunized with purified autolysin survived significantly longer than did control mice after intranasal challenge with strain D39. This study provides direct evidence that the pneumococcal autolysin contributes to virulence and identifies it as a potential vaccine antigen. PMID- 2568345 TI - Influence of a glycine or proline substitution on the functional properties of a 14-amino-acid analog of Escherichia coli heat-stable enterotoxin. AB - Analogs of Escherichia coli heat-stable enterotoxin (ST) differing in chain length or the presence of turn-forming residues were assessed for binding to receptors, activation of particulate guanylate cyclase, and stimulation of secretion in suckling mice. These analogs included the native 18-amino-acid peptide (ST), the 14-amino-acid carboxy terminus of this native peptide with a proline at position 12 (ST[5-18]proline), and the 14-amino-acid carboxy terminus in which the proline at position 12 was substituted with glycine (ST[5 18]glycine). Each analog bound to the receptor in a dose-dependent fashion, completely displacing [125I]ST in competitive binding assays. However, their potencies differed significantly: ST demonstrated the highest affinity (inhibition constant [Ki], 10(-9) M), followed by ST[5-18]proline (Ki, 10(-7) M) and ST[5-18]glycine (Ki, 10(-6) M). Similarly, these peptides maximally activated particulate guanylate cyclase and stimulated intestinal secretion in suckling mice. Their rank order of potency in these assays was similar to that described for receptor binding: ST greater than ST[5-18]proline greater than ST[5 18]glycine. These data demonstrate that the full peptide structure is not absolutely required for pharmacological, biochemical, or biological activity. However, the four amino-terminal residues contribute significantly to the potency of these peptides. In addition, the turn imposed by the proline residue at position 12 is not absolutely required for receptor occupancy or activation of the biochemical cascade that results in intestinal secretion. However, it significantly increases the potency of the toxin. These data illustrate the importance of primary and secondary structures to the biochemical, pharmacological, and physiological activities of the ST produced by E. coli. PMID- 2568346 TI - In vitro adherence of type 1-fimbriated uropathogenic Escherichia coli to human ureteral mucosa. AB - Type 1-fimbriated Escherichia coli isolated from patients with urinary tract infections adhered in vitro to the epithelial cell surface of an excised human ureter. The bacteria also adhered to a mucous coating and to Formalin-fixed human ureteral mucosa. D-Mannose strongly inhibited such adherence. The bacteria in their nonfimbriated phase lacked the ability to adhere. We concluded that type 1 fimbriae play a role, at least in part, in upper urinary tract infections in humans. PMID- 2568347 TI - Reversal of drug resistance by erythromycin: erythromycin increases the accumulation of actinomycin D and doxorubicin in multidrug-resistant cells. AB - Development of resistance to one type of lipophilic chemotherapeutic drug often leads to resistance to other, structurally unrelated, lipophilic drugs. This suggests that non-toxic lipophilic agents may interfere with and reverse drug resistance by saturating the pathway through which multidrug-resistant (MDR) cells protect themselves against cytotoxic drugs. The lipophilic antibiotic, erythromycin, can significantly reverse the resistance of MDR WEHI 164 murine fibrosarcoma cells to the chemotherapeutic drugs, doxorubicin and actinomycin-D. The MDR cells showed an approximately 10-fold higher expression of the P glycoprotein than the drug-sensitive parental cells from which the resistant cells were derived. The accumulation of actinomycin-D and doxorubicin was much lower in the drug-resistant cells than in the sensitive parental cells. The concentrations of erythromycin which reversed the drug resistance of the MDR cells increased the accumulation of actinomycin-D and doxorubicin in these cells to a level comparable to that observed in the sensitive parental cells. Our data suggest that erythromycin reverses drug resistance by saturating the drug-binding sites on the P-glycoprotein, thereby reducing the capacity of this protein to pump drugs out of resistant cells. Some of our MDR cells have also become more resistant to tumour necrosis factor (TNF). However, erythromycin did not reverse TNF resistance, suggesting that the mechanisms of multi-drug and TNF resistance are different. TNF did not influence drug accumulation in MDR cells. PMID- 2568348 TI - Differential effects of a new central adrenergic agonist--modafinil--and D amphetamine on sleep and early morning behaviour in young healthy volunteers. AB - Modafinil (CRL 40476) is a recently developed central alpha adrenergic agonist with vigilance-promoting properties. In a double-blind, placebo-controlled sleep laboratory study, its single-dose effects on objectively and subjectively evaluated sleep, morning awakening, and early morning behaviour were investigated and compared with amphetamine. Ten young healthy volunteers of both sexes spent 12 nights in the sleep laboratory: one adaptation night, one baseline night, five drug nights (100 mg and 200 mg modafinil; 10 mg and 20 mg d-amphetamine; placebo) and five subsequent washout nights. The drugs were administered in one week intervals according to a Latin square design. Somnopolygraphic investigations were performed between 22h30 and 06h00. Subjects received the drug orally half an hour before bedtime. A self-rating scale for sleep and awakening quality and early morning behavior was completed subsequent to the morning toilet. Thereafter, noopsychic and thymopsychic variables were evaluated utilizing a psychometric test-battery. Statistical analyses of objective sleep variables demonstrated that modafinil causes no significant changes as compared to a placebo. Sleep initiation remained unchanged after all of the drugs, while sleep maintenance was impaired dose-dependently after d-amphetamine. Thus, total sleep time and sleep efficiency decreased significantly after 20 mg d-amphetamine as compared to the placebo and modafinil. In regard to sleep architecture a reduction of sleep stage 2 and rapid eye movement-sleep occurred under d amphetamine while modafinil did not exhibit such an effect. Subjective sleep quality was significantly better after modafinil than after the reference compound. Subjective awakening quality and well-being in the morning did not show any significant findings. Furthermore, no differences were observed between the placebo and the other drugs concerning objective awakening quality (evaluated by psychometric tests). Critical flicker frequency increased significantly after 20 mg d-amphetamine as compared to the placebo. Pulse rate and evening and morning blood pressure remained unchanged. These data stress the necessity to differentiate between "vigility-increasing" properties of amphetamine and "vigilance-promoting" properties of modafinil. PMID- 2568349 TI - Ritanserin versus lorazepam: a double-blind, cross-over study of reaction times in healthy volunteers. AB - Ritanserin, a benzydrilen-piperidine derivative, with a potent, selective and long-lasting antagonist activity on serotonin type 2 receptors, has shown anxyolitic properties and a lesser sedative profile than benzodiazepines. Ritanserin and lorazepam at therapeutical doses, were compared in eight healthy volunteers in a double-blind, cross-over study in order to detect possible different impairments of performances. Before and during the treatment, all subjects were daily submitted to a test of rapidity and regularity of response to acoustic and visual stimuli by means of an electronic device. A visual analogue scale for the self-assessment of the drowsiness degree was also administered. Ritanserin did not modify reaction times, while lorazepam significantly prolonged them. The analysis of data within treatments significantly favoured ritanserin, while between treatments there was only a trend in favour of ritanserin. The analogical scale for concentration showed a significant reduction with lorazepam, whereas for drowsiness no alteration occurred with either drug. PMID- 2568350 TI - Double-blind placebo cross-over study of long-acting (chlordesmethyldiazepam) versus short-acting (lorazepam) benzodiazepines in generalized anxiety disorders. AB - Chlordesmethyldiazepam a long-acting benzodiazepine was compared with lorazepam a short-acting one in a double-blind placebo cross-over study against generalized anxiety disorders. Chlordesmethyldiazepam therapy was more effective than lorazepam. Clinical efficacy, drowsiness and insomnia seem well correlated with pharmacokinetic properties of these two benzodiazepines. These results further support the use of a long-acting benzodiazepine rather than a short-acting one as an anti-anxiety agent. PMID- 2568353 TI - Oral medication for pain relief. PMID- 2568352 TI - Influence of topical and systemic beta-blockers on tear production. AB - The influence of various beta-blockers on tear production in rabbits was studied by means of direct cannulation of the lacrimal gland excretory duct. Tear production was significantly decreased (p less than 0,001) after systemic (I.M.) administration of either selective beta 1-blockers (metoprolol, betaxolol), B2 selective (butoxamine) or non-selective ones (timolol, propranolol) or finally those with I.S.A. (oxprenolol), although differences in the size of tear production decreasing response of each beta-blocker were recorded. Bilateral topical application of timolol (50 microliters of 0,5% b.i.d. for three days), decreased also tear production significantly, while the response of betaxolol administered under the same regime was rather negligible. It is concluded that although beta 2 adrenergic receptors are mainly involved in tear production, the administration of selective beta 1-blockers in high doses disappears their selectivity, decreasing tear production too. PMID- 2568351 TI - Adrenergic responses in isolated bovine retinal resistance arteries. AB - The reactivity of bovine retinal resistance arteries (i.d. ca. 200 microns) to selective alpha 1-, beta- and combined alpha- and beta-adrenoceptor stimulation was studied in vitro using phenylephrine, isoproterenol and norepinephrine, respectively. The results indicate that only functional alpha 1-adrenoceptors, possibly in low numbers, are present in these vessels. PMID- 2568354 TI - Stimulation of vagal pulmonary C-fibers by a single breath of cigarette smoke in dogs. AB - Inhalation of cigarette smoke into the lower airway via a tracheostomy evokes immediate apnea, bradycardia, and systemic hypotension in dogs. These responses can still be evoked when conduction in myelinated vagal fibers is blocked preferentially by cooling but are abolished by vagotomy, suggesting that they are mediated by afferent vagal C-fibers. To examine this possibility, we recorded impulses in pulmonary C-fibers in anesthetized, open-chest dogs and delivered 120 ml cigarette smoke to the lungs in a single ventilatory cycle. Pulmonary C-fibers were stimulated within 1 or 2 s of the delivery of smoke generated by high nicotine cigarettes, activity increasing from 0.3 +/- 0.1 to a peak of 12.6 +/- 1.3 (SE) impulses/s, (n = 60); the evoked discharge usually lasted 3-5 s. Smoke generated by low-nicotine cigarettes evoked a milder stimulation in 33% of pulmonary C-fibers but did not significantly affect the overall firing frequency (peak activity = 2.2 +/- 1.1 impulses/s, n = 36). Hexamethonium (0.7-1.2 mg/kg iv) prevented C-fiber stimulation by high-nicotine cigarette smoke (n = 12) but not stimulation by right atrial injection of capsaicin. We conclude that pulmonary C-fibers are stimulated by a single breath of cigarette smoke and that nicotine is the constituent responsible. PMID- 2568355 TI - The function of Gp170, the multidrug resistance gene product, in rat liver canalicular membrane vesicles. AB - Gp170 (also known as P-glycoprotein) is a transmembrane glycoprotein which is overexpressed in multidrug-resistant tumor cells and is also found in the apical plasma membrane domain of several normal human and animal tissues. Gp170 has been postulated to function as an energy-dependent efflux pump for cytotoxic drugs. In rat liver, Gp170 is restricted to the bile canalicular domain of the plasma membrane. Canalicular membrane vesicles (CMV), but not sinusoidal membrane vesicles, contained a approximately 160-kDa protein which reacts with anti-Gp170 monoclonal antibody and manifest ATP-dependent [3H]daunomycin transport which is temperature dependent, osmotically sensitive, and saturable. Among several nucleotides, ATP was a potent stimulator of transport whereas non- or slowly hydrolyzable analogues (adenosin-5-O-(3-thiotriphosphate, adenyl-5-yl imidodiphosphate) were ineffective. ATP-dependent daunomycin transport was inhibited by cytotoxic drugs (vinblastine, vincristine, and adriamycin) and other drugs, such as verapamil and quinidine, which restore anti-cancer drug sensitivity in resistant cells. Inside-out CMV were separated from right side-out CMV by antibody-induced affinity density perturbation. Only inside-out CMV manifested ATP-dependent daunomycin transport. These results suggest that Gp170 is an ATP-dependent efflux pump which is responsible for the undirectional, energy-dependent transport of daunomycin and other drugs by rat liver into the bile. PMID- 2568356 TI - Cloning, sequence analysis, and permanent expression of a human alpha 2 adrenergic receptor in Chinese hamster ovary cells. Evidence for independent pathways of receptor coupling to adenylate cyclase attenuation and activation. AB - The gene encoding a human alpha 2-adrenergic receptor was isolated from a human genomic DNA library using a 367-base pair fragment of Drosophila genomic DNA that exhibited 54% identity with the human beta 2-adrenergic receptor and 57% identity with the human alpha 2-adrenergic receptor. The nucleotide sequence of a fragment containing the human alpha 2-receptor gene and 2.076 kilobases of untranslated 5' sequence was determined, and potential upstream regulatory regions were identified. This gene encodes a protein of 450 amino acids and was identified as an alpha 2-adrenergic receptor by homology with published sequences and by pharmacological characterization of the protein expressed in cultured cells. Permanent expression of the alpha 2-receptor was achieved by transfecting Chinese hamster ovary (CHO) cells which lack adrenergic receptors with a 1.5-kilobase NcoI-HindIII fragment of the genomic clone containing the coding region of the gene. The alpha 2-receptor expressed in CHO cells displayed pharmacology characteristic of an alpha 2 A-receptor subtype with a high affinity for yohimbine (Ki = 1 nM) and a low affinity for prazosin (Ki = 10,000 nM). Agonists displayed a rank order of potency in radioligand binding assays of para aminoclonidine greater than or equal to UK-14304 greater than (-)-epinephrine greater than (-)-norepinephrine greater than (-)-isoproterenol, consistent with the identification of this protein as an alpha 2-receptor. The role of the alpha 2-receptor in modulating intracellular cyclic AMP concentrations was investigated in three transfected cell lines expressing 50, 200, and 1200 fmol of receptor/mg membrane protein. At low concentrations (1-100 nM), (-)-epinephrine attenuated forskolin-stimulated cyclic AMP accumulation by up to 60% in a receptor density dependent manner. At epinephrine concentrations above 100 nM, cyclic AMP levels were increased up to 140% of the forskolin-stimulated level. Pertussis toxin pretreatment of cells eliminated alpha 2-receptor-mediated attenuation of forskolin-stimulated cyclic AMP levels and enhanced the receptor density dependent potentiation of forskolin-stimulated cyclic AMP concentrations from 3 to 8-fold. Potentiation of forskolin-stimulated cyclic AMP levels was also elicited by the alpha 2-adrenergic agonists, UK-14304 and para-aminoclonidine, and blocked by the alpha 2-adrenergic antagonist yohimbine, but not by the alpha 1-adrenergic antagonist prazosin or the beta-adrenergic antagonist propranolol.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2568357 TI - Molecular cloning of a Chinese hamster mitochondrial protein related to the "chaperonin" family of bacterial and plant proteins. AB - The complete cDNA sequence of a mitochondrial protein from Chinese hamster ovary cells, designated P1, which was originally identified as a microtubule-related protein (Gupta, R.S., Ho, T.K.W., Moffat, M.R.K., and Gupta, R. (1982) J. Biol. Chem. 257, 1071-1078), has been determined. The P1 cDNA encodes a protein of 60,983 Da including a 26-amino acid putative mitochondrial targeting sequence at its N-terminal end. The deduced amino acid sequence of Chinese hamster P1 shows 97% identity to the human P1 protein. Most interestingly, the amino acid sequences of mammalian P1 proteins show extensive sequence homology (42-60% identical residues and an additional 15-25% conservative replacements) to the "chaperonin" family of bacterial, yeast, and plant proteins (viz. groEL protein of Escherichia coli, hsp 60 protein of yeast, and ribulose-1,5-bisphosphate carboxylase subunit binding protein of plant chloroplasts) and to the 60-65-kDa major antigenic protein of mycobacteria and Coxiella burnetii. The homology between mammalian P1 and other proteins begins after the putative mitochondrial presequence and extends up to the C-terminal end. Furthermore, similar to the chaperonin family of proteins, P1 appears to exist in cells as a homooligomeric complex of seven subunits and shows ATPase activity. These observations strongly indicate that P1 protein is a member of the chaperonin family and that it may be involved in a similar function in mammalian cells. PMID- 2568358 TI - Coordinate regulation of 3-hydroxy-3-methylglutaryl-coenzyme A synthase, 3 hydroxy-3-methylglutaryl-coenzyme A reductase, and prenyltransferase synthesis but not degradation in HepG2 cells. AB - Human hepatoma HepG2 cells were used to demonstrate coordinate regulation of three enzymes of cholesterol synthesis under a variety of conditions. Addition of either delipidized serum and mevinolin or low density lipoprotein, 25 hydroxycholesterol, or mevalonic acid to HepG2 cells resulted in rapid changes both in the levels of the mRNAs and in the rates of synthesis of 3-hydroxy-3 methylglutaryl-coenzyme A (HMG-CoA) synthase, HMG-CoA reductase, and farnesyl pyrophosphate synthetase (prenyltranferase). In all cases, the changes in mRNA levels were paralleled by changes in the rates of specific protein synthesis. Pulse-chase techniques were used to determine the half-lives of all three proteins. Addition of low density lipoprotein to the media during the chase increased the rate of degradation of HMG-CoA reductase 4.6-fold but had no affect on the half-lives of HMG-CoA synthase or prenyltransferase. Therefore, we conclude that the coordinate regulation of these three enzymes under a variety of conditions occurs at the level of enzyme synthesis and not at the level of protein stability. PMID- 2568359 TI - An analog of lophotoxin reacts covalently with Tyr190 in the alpha-subunit of the nicotinic acetylcholine receptor. AB - Lophotoxin and lophotoxin analog-1 are natural diterpenes from coral that inhibit nicotinic acetylcholine receptors by covalent reaction with the acetylcholine recognition sites on the alpha-subunits. Although both toxins contain potentially reactive epoxides and alpha,beta-unsaturated aldehydes, the mechanism of their covalent reaction with the receptor is not known. The role of the alpha,beta unsaturated aldehyde in analog-1 was investigated by reduction of the aldehyde to an alcohol with [3H]NaBH4. The reduced [3H]analog-1 bound selectively and covalently to the alpha-subunit of the receptor. Covalent binding was inhibited by agonists and antagonists, but not by noncompetitive allosteric inhibitors. The apparent dissociation constant of the reduced [3H]analog-1 was approximately 1.5 x 10(-6) M. These results demonstrate that the alpha,beta-unsaturated aldehyde in analog-1 is not an absolute requirement for covalent reaction with the receptor. Receptors were treated with the reduced-[3H]analog-1, and the labeled alpha subunits were isolated by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis and digested with staphylococcal V8 protease. A labeled 20-kDa V8 protease fragment was purified by preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis and reverse-phase high performance liquid chromatography and subjected to sequence analysis. A peptide beginning at Ser173 was identified, and the label appeared in the 18th step corresponding to Tyr190. This assignment was confirmed by digestion of the labeled 20-kDa V8 protease fragment with cyanogen bromide, followed by purification of the labeled cyanogen bromide peptide on reverse-phase high performance liquid chromatography. A peptide beginning at Lys179 was identified, and the label appeared in the 12th step, again corresponding to Tyr190. Tyr190 may react with the coral toxin by nucleophilic addition at one of the carbons associated with an epoxide, and may form part of the alkylammonium-binding subsite of the acetylcholine recognition site. PMID- 2568360 TI - The action of a beta-adrenoreceptor agonist on gastric acid secretion in dogs. AB - 1. In conscious dogs with gastric fistulae and Heidenhain pouches, isoprenaline (0.75 microgram kg-1 h-1) brought about an increase in methacholine stimulated gastric acid and pepsin secretion from the innervated mucosa. 2. The increase in acid consisted for the most part of the replacement of the usual post-peak fade by a sustained plateau and of the replacement of the usual wait for the spontaneous peak by immediate stimulation of secretion. PMID- 2568361 TI - Organ-bath studies using the irreversible beta-adrenoreceptor antagonist bromoacetylalprenololmenthane (BAAM). AB - 1. The effects of the irreversible beta-adrenoreceptor antagonist bromoacetylalprenololmenthane (BAAM) were studied in isolated cardiac and uterine preparations from guinea-pigs and rats and in guinea-pig ileal preparations. 2. In the presence of BAAM (0.1-10 microM) concentration-effect curves to (-) isoprenaline were shifted to the right in a concentration-dependent manner in all cardiac and uterine tissues. Maximum responses to (-)-isoprenaline were unaffected by BAAM except in guinea-pig left atrial and in some guinea-pig uterine preparations; however, the reductions in the maximum responses were not concentration-dependent. The mean pKB values for BAAM in guinea-pig left atria, right atria, rat whole atria and rat uterus were 7.26, 7.24, 6.84 and 7.90 respectively. 3. In guinea-pig ileal preparations, BAAM (0.1-30 microM) relaxed contractions induced by K+, histamine and acetylcholine in a non-beta adrenoreceptor-related manner since relaxant responses were unaffected by propranolol (0.5 microM). In other tissues higher concentrations of BAAM (30-100 microM) elicited atrial standstill and depressed K+-induced contractions in uterine smooth muscle. 4. Treatment of tissues with BAAM (10-100 microM) followed by extensive wash-out increased the EC50 values for (-)-isoprenaline 21- to 83 fold. The maximum response to the catecholamine was unaffected by BAAM except in guinea-pig left atrial preparations following treatment with 100 microM BAAM. At these concentrations BAAM markedly increased the effective refractory period. 5. Concentration-effect curves for the partial agonist, oxymethylene-isoprenaline (OM-ISO) were shifted to the right 12- to 355-fold after pretreatment of tissues with BAAM (10-30 microM) followed by wash-out. The maximum response to OM-ISO was unaltered in guinea-pig and rat uteri and was reduced to a similar degree as observed with (-)-isoprenaline in guinea-pig left atria. 6. In general, the non selective beta-adrenoreceptor antagonist BAAM depressed maximum responses to beta adrenoreceptor agonists only in cardiac preparations and at concentrations which elicited depressant activity. On the basis of the present study, BAAM does not appear to be a suitable irreversible beta-adrenoreceptor antagonist for use in organ bath experiments. PMID- 2568362 TI - Interactions of the ulcerogen cysteamine with enteric inhibitory nerves and putative transmitters in the rat small intestine. AB - 1. The effects of the ulcerogen cysteamine (2-aminoethanethiol HCL) on spontaneous activity and evoked responses of rat isolated small intestine preparations were investigated in vitro. 2. Cysteamine induced concentration dependent relaxations of isolated segments of the rat duodenum, jejunum and ileum. These actions were manifest simultaneously in both the circular and longitudinal muscle layers, where the responses displayed a similar profile. 3. Treatment with tetrodotoxin (0.1 microM) or cold storage of individual preparations to prevent nerve-mediated responses abolished the effects of cysteamine. The presence of atropine (0.1 microM), propranolol (3.0 microM) and phentolamine (3.0 microM) in the bathing solution did not affect the cysteamine evoked relaxations, suggesting cysteamine was stimulating non-adrenergic, non cholinergic (NANC) intrinsic inhibitory nerves. 4. Applied cysteamine reversibly reduced GABA- and DMPP-evoked NANC nerve-mediated relaxations via actions unrelated to the receptors for these agents. 5. Methysergide-sensitive (direct) actions of 5-HT on the muscularis but not 5-HT neurally evoked responses were blocked by cysteamine. 6. It would appear that cysteamine has excitatory and inhibitory actions on enteric inhibitory nerves as well as specifically interfering with myogenic but not neural actions of 5-HT. PMID- 2568364 TI - Improved screening method for beta-blockers in urine using solid-phase extraction and capillary gas chromatography-mass spectrometry. AB - An improved screening method for beta-blockers in urine is proposed, involving enzymatic hydrolysis, solid-phase extraction and capillary gas chromatography mass spectrometry. Several extraction methods for beta-blockers, such as conventional liquid-liquid and solid-phase extraction procedures, have been evaluated for at least eight beta-blockers. Additionally, the gas chromatographic properties and mass fragmentation of the trimethylsilyl-trifluoroacetyl, trifluoroacetyl and cyclic n-butylboronate derivatives of beta-blockers have been compared and evaluated with respect to their efficiency for screening urine. The resulting screening method proved to be a specific and sensitive procedure, enabling these analytes to be detected and identified up to 48 h after the administration of a dosage, usually encountered in doping cases. PMID- 2568365 TI - Determination of picogram levels of a novel alpha 2-adrenergic receptor antagonist in plasma using solid-phase extraction and capillary gas chromatography with nitrogen-selective detection. PMID- 2568363 TI - Role of microtubules in polarized delivery of apical membrane proteins to the brush border of the intestinal epithelium. AB - Colchicine- and vinblastine-induced depolymerization of microtubules (MTs) in the intestinal epithelium of rats and mice resulted in significant delivery of three apical membrane proteins (alkaline phosphatase, sucrase-isomaltase, and aminopeptidase N) to the basolateral membrane domain. In addition, typical brush borders (BBs) occurred at the basolateral cell surface, consisting of numerous microvilli that contained the four major components of the cytoskeleton of apical microvilli (actin, villin, fimbrin, and the 110-kD protein). Formation of basolateral microvilli required polymerization of actin and proceeded at glycocalyx-studded plaques that resembled the dense plaques located at the tips of apical microvilli. BBs from the basolateral membrane became internalized into BB-containing vacuoles which served as recipient organelles for newly synthesized apical membrane proteins. The BB vacuoles fused with each other and finally were inserted into the apical BB. Polarized distribution of Na+,K+-ATPase, a basolateral membrane protein, was not affected by drug-induced depolymerization of MTs. These observations indicate that Golgi-derived carrier vesicles (CVs) containing apical membrane proteins are vectorially guided to the apical cell surface by a retrograde transport along MTs. MTs are uniformly oriented towards a narrow space underneath the apical terminal web (termed subterminal space) that contains MT-organizing properties and controls polarized alignment of MTs. In contrast to apical CVs, targeting of basolateral CVs appears to be independent of MTs but demands a barrier at the apical membrane domain that prevents basolateral CVs from apical fusion (transport barrier hypothesis). PMID- 2568366 TI - Determination of the beta-adrenoceptor blocking drug B24/76 in serum by high performance liquid chromatography with fluorimetric detection after pre-column dansylation. PMID- 2568367 TI - Stability of serum gamma-glutamyltranspeptidase fractionated by gel permeation using high-performance liquid chromatography and fast protein liquid chromatography. PMID- 2568368 TI - Detection of piluslike structures on clinical and environmental isolates of Vibrio vulnificus. AB - Twenty clinical isolates of Vibrio vulnificus were compared with 10 environmental strains by using electron microscopy and agglutination assays with human erythrocytes, guinea pig erythrocytes, and Saccharomyces cerevisiae. In addition, the isolates were tested for ability to adhere to the human epithelial cell lines HEp-2 and A549. When examined by electron microscopy, 16 (80%) of the 20 clinical isolates demonstrated the presence of piluslike structures; the composition of the bacterial populations ranged from 0 to 68% piliated cells. In contrast, only 3 (30%) of the 10 environmental isolates were piliated, with a range from 0 to 16% piliated cells. A significant association between the presence of piliated cells and the isolate source was found (P less than 0.05). None of the 30 strains agglutinated erythrocytes or yeast cells. V. vulnificus adherence results obtained with HEp-2 cells showed 10 (50%) of 20 clinical isolates and 0 (0%) of 10 environmental isolates with averages of greater than 10 adherent bacteria per cell, demonstrating a correlation between attachment and the isolate source (P less than 0.05). Selected strains were tested to determine whether methyl alpha-D mannopyranoside, fructose, or alpha-L-(-)-fucose would inhibit bacterial adherence to HEp-2 cells. Multiple patterns of adherence inhibition were observed. Adherence to A549 cells showed 8 (40%) of 20 clinical isolates and 0 (0%) of 10 environmental strains with averages of greater than 10 adherent bacteria per cell. A statistical association between attachment and the isolate source was demonstrated (P less than 0.05). These data suggest that the presence of piluslike structures and the ability to adhere to human epithelial cell lines may be more closely associated with V. vulnificus isolates from clinical specimens than with environmental strains. PMID- 2568369 TI - Sixth nerve palsies, temporal artery biopsy, and necrotizing vasculitis. AB - A 79-year-old man with diplopia and weakness was found to have necrotizing vasculitis consistent with polyarteritis nodosa on temporal artery biopsy. Although he had no evidence of visceral involvement from vasculitis, he responded well to corticosteroid therapy. This case illustrates a nosological problem that can occur when dealing with a disease that has diverse clinical manifestations and nonspecific laboratory findings. Moreover, this case demonstrates the value of including a branch artery segment when biopsying the temporal artery. PMID- 2568370 TI - Bed nucleus of the stria terminalis: cytoarchitecture, immunohistochemistry, and projection to the parabrachial nucleus in the rat. AB - The bed nucleus of the stria terminalis (BST) sends a dense projection to the parabrachial nucleus (PB) in the pons. The BST contains many different types of neuropeptidelike immunoreactive cells and fibers, each of which exhibits its own characteristic distribution within cytoarchitecturally distinct BST subnuclei. Corticotropin releasing factor (CRF)-, neurotensin (NT)-, somatostatin (SS)-, and enkephalin (ENK)-like immunoreactive (ir) neurons are particularly numerous within areas of the BST that project to the PB. In this study, we use the combined retrograde fluorescence-immunofluorescence method to determine whether neurons in the BST that project to the PB contain these immunoreactivities. After Fast Blue injections into PB, retrogradely labeled neurons were numerous throughout the lateral part of the BST, particularly in the dorsal lateral (DL) and posterior lateral subnuclei. Retrogradely labeled neurons were also present in the preoptic, ventral lateral, and supracapsular BST subnuclei and in the parastrial nucleus. Many of the CRF-ir, NT-ir, and SS-ir neurons in DL were retrogradely labeled. A few double-labeled cells of each type were also found in the posterior lateral, ventral lateral and supracapsular BST subnuclei ENK-ir neurons were never retrogradely labeled. Our results show that BST neurons that project to the PB stain for the same neuropeptides as those in the central nucleus of the amygdala (CeA) that project to the PB, demonstrating further the close anatomical relations between these two structures. PMID- 2568371 TI - Structure, afferent innervation, and transmitter content of ganglia of the guinea pig gallbladder: relationship to the enteric nervous system. AB - Although a well-developed plexus of nerves and ganglia is known to be present in the wall of the gallbladder, little has previously been learned about the function or organization of this innervation. The current study was undertaken in order to evaluate the hypothesis that the ganglionated plexus of the gallbladder is analogous to elements of the enteric nervous system (ENS). The ganglionated plexus of the gallbladder was found to resemble closely the submucosal plexus of the small intestine in its organization into two irregular anastomosing and interwoven networks of ganglia, in the numbers of neurons per ganglion, and in the manifestation of histochemically demonstrable acetylcholinesterase activity in virtually all ganglion cells. In common with enteric ganglia, laminin immunoreactivity was observed to be excluded from the interiors of gallbladder ganglia, which were surrounded by a periganglionic laminin-immunoreactive sheath. As in the submucosal plexus, intrinsic substance P-, vasoactive intestinal polypeptide (VIP)-, and neuropeptide Y (NPY)-immunoreactive neurons were seen in the ganglionated plexus of the gallbladder. Extrinsic nerves in the gallbladder that degenerated following chemical sympathectomy with 6-hydroxydopamine (6 OHDA), and which contained NPY, tyrosine hydroxylase (TH), and dopamine-beta hydroxylase (DBH) immunoreactivities, formed a perivascular plexus closely associated with blood vessels. Endogenous catecholamines could also be demonstrated in these perivascular nerves by aldehyde-induced histofluorescence. In addition to perivascular nerves, paravascular nerve bundles were observed that were loosely associated with vessels, did not degenerate following administration of 6-OHDA, and contained NPY immunoreactivity. Other paravascular nerves, probably visceral sensory axons, coexpressed substance P and calcitonin-gene related peptide (CGRP) immunoreactivities. The ganglionated plexus of the gallbladder resembled enteric ganglia in having intrinsic 5-hydroxytryptamine (5 HT)-immunoreactive cells and highly varicose nerve fibers. The 5-HT immunoreactive gallbladder axons were, like those of the gut, resistant to 6 OHDA, and separate from fibers that expressed TH immunoreactivity. Differences between the ganglionated plexus of the gallbladder and enteric ganglia of the small intestine included in the gallbladder are 1) the presence of TH immunoreactive cells that contain an endogenous catecholamine, but not DBH; 2) DBH-immunoreactive neurons, some of which coexpress substance P immunoreactivity, but which contain neither a catecholamine nor TH immunoreactivity; 3) an apparent absence of CGRP-immunoreactive cell bodies.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2568372 TI - Vasopressin- and proctolin-like immunoreactive efferent neurons in blowfly abdominal ganglia: development and ultrastructure. AB - In the neural sheath of the fused thoracicoabdominal ganglia of the blowfly Calliphora erythrocephala, extensive neurohaemal areas can be seen in the electron microscope. A separate set of neurohaemal areas located in the sheath of the lateral abdominal nerve roots contain neural terminals of at least three morphological types. To determine which bioactive substances are stored and possibly released from the neurons supplying these neurohaemal areas, we applied a large number of antisera raised against different neuropeptides of invertebrate and mammalian type. Antisera to two types of neuropeptides react with neurons innervating the sheath of the abdominal nerve roots: antisera to lysine vasopressin and proctolin. There are only 14-24 vasopressin-like immunoreactive (VPLI) neurons in the entire nervous system of Calliphora. These are all restricted to a bilateral cluster in the fused abdominal ganglia. From this cluster, the neurohaemal areas in abdominal nerve roots are supplied. Proctolin like immunoreactivity (PLI) can be seen in a large number of neurons in the nervous system of blowflies. The supply of PLI terminals to the abdominal nerve roots is from 12 to 14 neurons in a bilateral cluster of abdominal PLI neurons. It is clear from light- and electron-microscopic immunocytochemistry that the two antisera label two separate populations of neurons that form overlapping terminals in the neural sheath. The immunoreactive terminals are located just below the permeable acellular basal lamina of the neural sheath. Hence, it is likely that at least two different bioactive peptides can be released neurohormonally into the circulation. An additional set of four efferent PLI neurons send axons into the medial abdominal nerve. These do not form neurohaemal terminals in the nerve root, but may innervate the hindgut. Also in the larval nervous system, VPLI and PLI neurons can be recognized. In the larva, the peptide containing neurons are segmentally arranged. The 14 larval VPLI neurons supply segmental abdominal nerves with axons that run inside the nerves to their targets. During metamorphosis, the segmental nerves fuse and the VPLI axons invade the neural sheath where they arborize and form varicose terminals. About the same number of PLI neurons could be detected in the abdominal ganglia of larval and adult flies. Only for a set of four caudal PLI neurons could efferent axons be traced in the larva. These axons run inside the medial abdominal nerves. The same four PLI neurons, with the same axonal projections, can be recognized in the adults. PMID- 2568373 TI - The chicken major histocompatibility complex in disease resistance and poultry breeding. AB - Numerous studies confirm that genes in the chicken major histocompatibility complex exert major genetic control over host resistance to autoimmune, viral, bacterial, and parasitic diseases. Examples of major histocompatibility complex associations with traits of growth and reproduction in the chicken are also available. Thus, the major effects of the major histocompatibility complex on the economically important traits of disease resistance, growth, and reproduction make the major histocompatibility complex a valuable subject for intensive analysis in agricultural species. This paper examines, as a model for integration of genetics and immunology, the research on the chicken major histocompatibility complex, which confirmed its role in genetic control of disease resistance, focusing on Marek's disease, a virally induced cancer. Current knowledge of associations of the chicken major histocompatibility complex with specific disease resistance, immune response, and other economic traits are selectively reviewed. Use of major histocompatibility complex typing in the poultry industry, including speculation about future applications, is presented. PMID- 2568375 TI - Pentagastrin stimulation of calcitonin in pheochromocytoma does not always indicate multiple endocrine neoplasia type II. AB - The diagnosis of pheochromocytoma in a 48-year-old man was confirmed by elevated catecholamine secretion and a left adrenal mass on computerized tomography. Because of a plausible family history for Multiple Endocrine Neoplasia Type II, a calcitonin level was determined which was elevated, and pentagastrin stimulation caused a 235% increase. These findings normalized following surgical removal of the single adrenal tumor. It is concluded that pentagastrin stimulation of calcitonin is not necessarily diagnostic of medullary thyroid carcinoma, and such a response in a patient presenting with pheochromocytoma may not indicate underlying Multiple Endocrine Neoplasia Type II. PMID- 2568374 TI - Improvement of infiltrative ophthalmopathy in parallel with decrease of thyroid stimulating antibody (TSAb) activity in two patients with hypothyroid Graves' disease. AB - Two patients with primary hypothyroidism associated with infiltrative ophthalmopathy without previous history of hyperthyroidism are presented. Anti TSH receptor antibodies (TRAb) were detected by radioreceptor assay (TBII), and unexpectedly their biological activity was not of a blocking (TSBAb), but of a thyroid-stimulating type (TSAb). After the initiation of levothyroxine therapy, the TBII and TSAb activities both decreased gradually with normalization of the elevated TSH level. The inflammatory eye signs improved strikingly in parallel with decrease of these antibody activities. These data indicate that (1) TRAb in primary hypothyroidism do not always show TSAb activity, (2) the decrease in TRAb following levothyroxine therapy in these patients appeared to correlate with suppression of TSH, (3) changes in infiltrative ophthalmopathy were associated with that of TSAb even in primary hypothyroidism, and (4) the hypothyroidism in these patients is justifiably diagnosed as "hypothyroid Graves' disease". TSAb might be somewhat related to the pathogenesis of ophthalmopathy in autoimmune thyroid diseases. PMID- 2568377 TI - Guidelines for the dosage of neuroleptics. II: Changing from daily oral to long acting injectable neuroleptics. AB - When changing from daily oral to long acting injectable neuroleptic therapy, it is essential to choose a dose that will maintain clinical efficacy without increased adverse reactions. Information on pharmacokinetics is needed to achieve this. The authors present guidelines based on this information and observe that previous recommendations sometimes differ substantially from these guidelines. Whether these guidelines will improve the results of long term treatment of psychotic patients remains to be determined. PMID- 2568376 TI - Dystonia presenting as upper airway obstruction. AB - Dystonic extrapyramidal reactions due to neuroleptic medications are a common entity in emergency medicine. However, it is not widely reported that dystonia can be associated with airway compromise and death. We describe a case of upper airway obstruction due to a dystonic reaction to haloperidol, present radiographic evidence of airway compromise, and explain the physiologic mechanisms involved. PMID- 2568378 TI - Guidelines for the dosage of neuroleptics. I: Chlorpromazine equivalents of orally administered neuroleptics. AB - Chlorpromazine (CPZ) equivalents are often used as a relative measure of the antipsychotic potency of neuroleptics. We review the CPZ equivalents of 33 neuroleptics and illustrate why imprecisions or discrepancies will be difficult to eliminate from these values. We nevertheless underline that CPZ equivalents can be clinically useful, since they facilitate the choice of doses of different neuroleptics that should induce comparable antipsychotic effects. PMID- 2568379 TI - Somatostatin gene expression in the thymus gland. AB - A complex pattern of interactions appears to exist between the immune and neuroendocrine systems. Recently, vasopressin, oxytocin and vasoactive intestinal peptide have been isolated from the thymus. Using a rat somatostatin antisense RNA probe we have demonstrated expression of the somatostatin gene in the rat thymus. Furthermore, we have shown that the levels of thymic somatostatin mRNA exhibit a bell-shaped response to dexamethasone administration. Lipocortin I and II antisense RNA probes have been used as a positive control for the effects of the dexamethasone. We would suggest that somatostatin acts in the thymus in a paracrine mode to modulate T lymphocyte development. PMID- 2568380 TI - CD44 contributes to T cell activation. AB - We demonstrate here that the CD44 molecule, which mediates lymphocyte adhesion to high endothelial venules (HEV), is also involved in the delivery of an activation signal to the T cell. We have produced a CD44 mAb (H90) which is able to block the binding of lymphocytes to high endothelial venules. H90 had no effect on [3H]TdR incorporation of whole PBL stimulated by lectins, allogeneic cells, or CD3 mAb in the soluble phase; in contrast, it strongly increased [3H]TdR incorporation of PBL stimulated by CD2 pairs of mAb or by CD3 mAb linked to the plastic culture plates, when purified T cells were used, H90 mAb could efficiently induce them to proliferate after a primary signal of activation delivered via cross-linked CD3 or via CD2, an effect mediated by Il-2 synthesis and Il-2R expression. Thus, the effect of H90 mAb resembles the mitogenic effect of CD28 "9.3" mAb. However, several results show that CD28 and CD44 mediate different signals to the T cells: i) in contrast to CD28 mAb, CD44 mAb cannot complement the signal delivered by a soluble CD3 mAb, lectins, or PMA; ii) CD44 mAb, at the difference of CD28 mAb, cannot induce CD3+ thymocytes to proliferate in conjunction with a first signal provided via cross-linked CD3 or via CD2; iii) F(ab) fragments of H90 were efficient, whereas divalent fragments of CD29 9.3 mAb are required to produce activation signals; and iv) CD44 and CD28 mAb produce a very strong synergistic effect on T cell proliferation. These results fit with previous ones showing that endothelial cells can play the role of accessory cell in T cell activation and that a hierarchy of signaling can be delivered to T cells via CD3 and CD2. PMID- 2568381 TI - The effect of anti-intercellular adhesion molecule-1 on phorbol-ester-induced rabbit lung inflammation. AB - The role of the CD18 complex of leukocyte glycoproteins in adhesion-dependent functions of human leukocytes in vitro has been well documented. A ligand, intercellular adhesion molecule-1 (ICAM-1), for at least one member of the CD18 complex has been identified. This molecule is inducible on many cell types including vascular endothelium and keratinocytes by inflammatory mediators such as IL-1, TNF, and IFN-gamma. ICAM-1 has been shown to mediate, in part, the in vitro adhesion of lymphocytes and neutrophils to endothelial cells expressing ICAM-1. In the present study we have shown that mAb's to the human CD18 complex and to human ICAM-1 cross react with rabbit cells and that both anti-CD18 and anti-CD11b but neither anti-CD11a nor anti-ICAM-1 mAb's inhibit neutrophil migration, an adhesion-dependent function, in vitro. Pretreatment of rabbits with anti-CD18 and anti-ICAM-1 but not anti-CD11a mAb inhibited by greater than 60% neutrophil migration into PMA-induced inflamed rabbit lungs. This effect of anti ICAM-1 mAb on pulmonary neutrophil influx after PMA injection has important implications. Specifically, that ICAM-1 can function as a ligand for CD18 and can mediate, at least in part, the migration of neutrophils to inflammatory sites. PMID- 2568382 TI - The endocrine pancreas in patients with insulinomas. An immunocytochemical and ultrastructural study of the nontumoral tissue with morphometrical evaluations. AB - The nontumoral endocrine pancreas was studied immunocytochemically and ultrastructurally in 12 patients with isolated insulinomas. Changes affecting hormone content and secretion of B-, A-, and D-cells were found in the islets of insulinoma-bearing patients when compared with controls, in terms of a significant decrease of the insulin-immunoreactive tissue areas and an increase in the glucagon- and somatostatin-immunoreactive ones. Conversely, only in two of the patients examined were PP-immunoreactive tissue areas augmented. Diffuse ducto-endocrine proliferation (nesidioblastosis) was also a common feature in the tumor-associated pancreas. Both morphometry and qualitative features revealed that islet cell hyperplasia occurs in the presence of insulinoma. Ultrastructural examination revealed that the functional activity of B-cells is substantially depressed in the insulinoma-bearing patients, whereas it is maintained or even enhanced in the other cell types. The islet content in immunoreactive insulin decreases along with duration of hypoglycemic symptoms. The present findings indicate that, in the presence of an insulinoma, the endocrine pancreas undergoes changes that can be regarded as an adaptive response to the chronic excess of insulin and are possibly responsible for the patients' postoperative clinical course. PMID- 2568383 TI - [Neurotropic fungal metabolites]. PMID- 2568384 TI - [Studies on developmental changes in rat pancreatic endocrine system during perinatal period]. AB - During the perinatal development of rats, pancreatic endocrine cells (B, A and D cells) were quantitated morphometrically and plasma insulin, glucagon and somatostatin were measured. Moreover, intrauterine growth retardation (IUGR) rat fetuses were induced by uterine artery ligation and at the 21st day of gestation their volume density of pancreatic endocrine cells and plasma hormone levels were compared with that of normal fetuses. At the 16th day A cells were more numerous than B cells. But after then, the volume density of B cells increased rapidly. Plasma insulin also increased in the fetal period and was very high at the late fetal day. Just after birth plasma insulin decreased immediately and plasma glucagon increased and reached a very high peak. The volume density of D cells was much lower than that of the other cell types, and plasma somatostatin did not change remarkably throughout the perinatal period. In the IUGR rat fetuses the volume density of B cells was significantly lower than that of controls. In addition, plasma insulin was lower in the IUGR group, whereas plasma glucagon was higher. These results suggest that the pancreatic endocrine system, especially insulin and glucagon play some important roles in fetal development and postnatal metabolic changes. PMID- 2568385 TI - [Hereditary factors in Takayasu arteritis--III. Polymorphism of human complements]. PMID- 2568386 TI - [A case of systemic lupus erythematosus with lupus nephritis occurring in Crohn's disease]. PMID- 2568387 TI - On-site work evaluations: desensitisation for avoidance reactions following severe hand injuries. AB - This study examined on-site work evaluations as an environmental exposure strategy to promote return to work in 15 recalcitrant patients who had failed to benefit from established methods of reducing post-traumatic stress disorder symptoms. Following the on-site work evaluation, 87% of these patients were able to use visualisation of the work setting to further desensitize themselves and returned to work within the next eight weeks. All 87% have continued to be employed at six and twelve month follow-ups. This approach holds promise for assisting patients with hand injuries who develop post-traumatic stress disorder and fail to respond to traditional psychological strategies. PMID- 2568388 TI - [Detection of triple X syndrome during a familial inquiry for hemophilia A]. AB - In order to give a genetic counsel to the mother and the two twin-sisters of an Hemophilia A boy, a familial investigation has been carried out. The study with the restriction fragment length polymorphism probes together with the hemostasis screening enabled to specify the status of each member of the family with respect to hemophilia A gene. This investigation evidenced the presence of two maternal alleles and one paternal allele in one of the twin-sisters. This led us to assume that she had a chromosomal abnormality in spite of her normal phenotype. The 47, XX, +(X) result of her karyotype confirmed this assumption. Therefore an accurate genetic counsel was provided to the members of this family and more particularly to the triple X subject. PMID- 2568389 TI - The importance of cnidarian synapses for neurobiology. AB - Despite being the most primitive organisms to possess a nervous system, cnidarians afford rare opportunities for studying various, general aspects of chemical synaptic transmission. This is made possible by the unique organization of their nervous systems and by the fact that in certain species the neurons and synapses are readily accessible for intracellular recordings and voltage clamp. The results obtained from such studies are summarized here, with particular emphasis on work with two species, Cyanea capillata (Scyphozoa) and Polyorchis pennicilatus (Hydrozoa). The potential of these preparations for providing additional data is also discussed. PMID- 2568390 TI - Proctolin: a review with emphasis on insects. AB - The distribution, physiological role, mode of action, and pharmacology of the pentapeptide neuroregulator proctolin are reviewed, with special emphasis on insects. Whereas proctolin is distributed extensively throughout arthropods, its presence in molluscs, annelids, or chordates is not well established. In the arthropods, proctolin acts as a neuromodulator and possibly as a neurohormone. It does not appear to function as a conventional neurotransmitter. Two model proctolinergic systems are highlighted: motor control of the visceral muscles of the locust oviduct and of the skeletal muscles of the locust ovipositor. In these preparations proctolin is a cotransmitter acting to enhance neuromuscular transmission and muscular contraction. The mode of action of proctolin is not well understood, although the second messengers cAMP, phosphatidyl inositol, and calcium have been implicated in various systems. Pharmacologically, the proctolin receptor has been examined with structure/activity studies, and the effects of a variety of amino acid substitutions and deletions of the pentapeptide are described. It is unfortunate that no specific antagonists of the proctolin receptor appear to be available and that no receptor-binding studies have been reported. The prospects are good for advances in our understanding of modulatory mechanisms, since proctolin appears to be emerging as the model for studies of this type. PMID- 2568391 TI - Use of aerial color infrared photography as a survey technique for Psorophora columbiae oviposition habitats in Texas ricelands. AB - This study investigated the possibility of using aerial color infrared (CIR) photography as a survey technique for egg populations of Psorophora columbiae within a riceland agroecosystem. To accomplish this, eight photographic missions were flown over study fields during a variety of seasons and at various altitudes. Assessment of resulting photographic data indicates features reported in the literature as being attractive for oviposition by Ps. columbiae (i.e., rice field levees, tire tracks, ditches and low areas) can be readily detected on aerial CIR photographs. Features associated with Ps. columbiae oviposition sites were easily visible on photographs at scales as small as 1:42,000. PMID- 2568393 TI - Correlation between wing length and protein content of mosquitoes. AB - Wing length and protein content were compared in individual specimens of male and nonbloodfed, nongravid female mosquitoes. The following groups were tested: field collected and colonized Aedes aegypti, field-collected Culex nigripalpus and Aedes vexans, and colonized Culex quinquefasciatus and Psorophora columbiae. The correlation coefficient varied from 0.91 in Ae. aegypti males to 0.98 in Ae. vexans females, and in each group the P value was less than 0.001. This close correlation suggests that both wing length and protein measurement are reliable expressions of size in adult mosquitoes. PMID- 2568392 TI - Effects of weathering on fabrics treated with permethrin for protection against mosquitoes. AB - Permethrin-impregnated and untreated fabrics were evaluated for their toxic and repellent effects against Anopheles stephensi and Aedes aegypti after both types of fabrics were subjected to accelerated weathering for 9 weeks, under a simulated wet/tropical environment. The toxic (knockdown) effect of permethrin impregnated fabrics against both species of mosquitoes diminished rapidly after 1 week compared to the repellent effect. After 6 weeks of weathering, the remaining low amounts of permethrin provided fair protection from mosquito bites; however, no knockdown was observed at those levels. Permethrin-treated fabric was effective in providing protection from mosquito bites and appears to be a means of attenuating both the nuisance effects and, possibly, disease transmission by mosquitoes. PMID- 2568394 TI - Studies on the seasonality of Culiseta inornata in Kern County, California. AB - The seasonal abundance of adult Culiseta inornata was markedly bimodal in the San Joaquin Valley of Kern County, California, with minima observed during both summer and midwinter. Larvae were abundant in most surface water habitats during winter, but could not be found during summer. The occasional collection of females during summer indicated the persistence of an adult population. The midwinter decrease in adult abundance was attributed to the progressive mortality of the autumnal cohort and delayed emergence due to cold water temperature. Reproductive diapause was not induced experimentally when field or laboratory populations were exposed as larvae, pupae or adults to simulated summer or winter photoperiod and temperature regimens. In comparison, Culex tarsalis readily entered a winter diapause when concurrently exposed to simulated winter conditions. The aestivation, and perhaps hibernation, of reproductively quiescent females makes Cs. inornata theoretically attractive as a maintenance host of encephalitis viruses, while the bimodal seasonality of host-seeking activity defines periods when Jamestown Canyon virus may be transmitted horizontally. PMID- 2568395 TI - Biosystematics of larval movement of Central American mosquitoes and its use for field identification. AB - Means of locomotion of 48 species of larval mosquitoes was observed using cinematography in Panama, Honduras and neotropical Mexico. General observation led to a classification of movement into path, frequency, position and mechanism. Examination of high speed film sequences (64 frames per second) revealed that all species use the same basic mechanism of flexing, which consists of a power and a recovery phase. The entire flexing cycle is a modification of undulatory propulsion commonly observed in other animals (e.g., snakes, ceratopogonid larvae). Variations on the basic patterns of mosquito larval flexing mainly concern the speed of the power stroke and the degree of sinusoid curvature prior to the power stroke. Four basic patterns of larval flexing were discerned: irregular, sinuous, semisymmetric and anopheline. Some taxonomic groups appear to use one pattern of flexing exclusively or with few exceptions. Examples include Culex (Melanoconion) with sporadic irregular flexing, Cx. (Culex) with sustained irregular flexing, Haemagogus with slow sinuous flexing and Anopheles with anopheline flexing. Other groups (e.g., Aedes (Howardina), Cx. (Carrollia), Deinocerites, and Wyeomyia (Wyeomyia] use a number of patterns of flexing. Observation of flexing and other aspects of larval movement can be an important addition to geographical, habitat and morphological considerations in field identifications. PMID- 2568396 TI - The runnelling method of habitat modification: an environment-focused tool for salt marsh mosquito management. AB - Traditional methods of managing salt marsh mosquitoes focus primarily on maximizing the reduction of mosquito populations, with minimizing environmental impact as a secondary consideration. An environment-oriented approach to salt marsh management for mosquito control, runnelling, is described and compared with other forms of habitat modification such as ditching and Open Marsh Water Management (OMWM). Runnelling alters the salt marsh as little as possible while causing significant reductions in mosquito numbers. The effect of runnelling on the environment was monitored via the following variables: water table level, substrate characteristics (moisture, salinity and pH), vegetation (height and density of each Sporobolus virginicus) and the numbers of mosquito larvae. Runnelling had a statistically significant effect on only two of the seven variables. These were the height of Sporobolus, which increased near runnels, and the number of mosquito larvae, which decreased. The main difference between ditching, OMWM and runnelling lies in the magnitude of the habitat modification. Ditching involves the greatest alteration to the marsh, and runnelling the least. Consequently, runnelling has a smaller effect on the estuarine environment as a whole than does either ditching or OMWM. PMID- 2568397 TI - Suppression of larval Aedes aegypti populations in household water storage containers in Jakarta, Indonesia, through releases of first-instar Toxorhynchites splendens larvae. AB - Weekly releases of first-instar Toxorhynchites splendens larvae were made in household water storage containers in a neighborhood in Jakarta, Indonesia, between April 1987 and April 1988. A single larva was placed in each container surveyed. Forty-one percent of all containers in the treatment area were treated each week and the average container was treated once every 2.4 weeks. Aedes aegypti populations were suppressed but not controlled by treatment. It is hypothesized that first-instar Toxorhynchites larvae were poor control agents due to their inability to withstand periods of starvation and to their accidental removal from containers during the act of water consumption. PMID- 2568398 TI - Mosquito larval development in container habitats: the role of rotting Scirpus californicus. PMID- 2568399 TI - Efficacy of Duplex and Vectobac against Psorophora columbiae and Anopheles quadrimaculatus larvae in Arkansas ricefields. PMID- 2568400 TI - A mechanical aspirator for safe transfer of arbovirus-infected mosquitoes within containment chambers. PMID- 2568401 TI - A computer program to calculate pesticide concentrations for mosquito larval susceptibility tests. PMID- 2568402 TI - Purification and characterization of prosomatostatin from rat brain. AB - A high molecular weight somatostatin-immunoreactive polypeptide, presumably prosomatostatin, was purified from rat brain and characterized. Purification steps included extraction with 2 M acetic acid, precipitation of contaminating proteins at pH 6.5, Sephadex G-50 chromatography, immunoaffinity chromatography, and HPLC steps (size exclusion and reversed-phase HPLC). The protein was purified more than 30,000-fold. It is heat stable. Sodium dodecyl sulfate-gel electrophoresis and immunoblotting revealed one major immunoreactive band of approximately 13,000 molecular weight which roughly corresponds to the size of prosomatostatin as derived from its DNA sequence. Isoelectric focusing and two dimensional sodium dodecyl sulfate-gel electrophoresis gave a single immunoreactive spot at a pI of 5.4. The polypeptide did not bind to concanavalin A or to wheat germ lectin columns, suggesting lack of N-glycosylation in the molecule. Regional distribution of prosomatostatin varied between 6%, 10%, and 18% of total immunoreactivity in the brainstem, cortical areas, and striatum, respectively. PMID- 2568403 TI - Spontaneous glutamate release by a "fibrous"-like cerebellar astroglial cell clone. AB - To investigate the role of astrocytes in the metabolism of glutamate, the neurotransmitter of the granule cells of the cerebellar cortex, we have analyzed various parameters related to the synthesis of glutamate in astroglial cell clones that may be the in vitro counterparts of the cerebellar astrocytes. The "fibrous"-like clone spontaneously released large quantities of glutamate, even in the absence of glutamine in the culture medium, but did not release alanine. In contrast, the "Golgi-Bergmann"-like cells released alanine but not glutamate, whereas the "velate-protoplasmic"-like astrocytes released little glutamate and alanine. However, the glutamate oxaloacetate transaminase and glutamate pyruvate transaminase activities of the three astroglial cell lines, measured in the direction of glutamate synthesis, were comparable. In addition, the "velate protoplasmic" and "Golgi-Bergmann" clones did not consume glutamine present at 2 mM in the culture medium. These data suggest that the different types of in vivo cerebellar astrocytes may have distinct roles regarding glutamate-glutamine metabolism. PMID- 2568404 TI - In vitro reactivation of rat cortical tryptophan hydroxylase following in vivo inactivation by methylenedioxymethamphetamine. AB - The activity of tryptophan hydroxylase (EC 1.14.16.4) from rat brain was significantly decreased 1 h following a single systemic injection of 3,4 methylenedioxymethamphetamine (MDMA) when assessed ex vivo by radioenzymatic assay or in vivo by the quantitation of 5-hydroxytryptophan accumulation following central L-aromatic amino acid decarboxylase inhibition. Recovery of enzymatic activity in vivo, which occurred within 24 h of low-dose MDMA treatment, appeared not to involve synthesis of new enzyme protein, because the return of enzymatic activity was not prevented by prior cycloheximide. Acutely MDMA-depressed cortical tryptophan hydroxylase activity could be completely restored in vitro by a prolonged (20-24 h) anaerobic incubation in the presence of dithiothreitol and Fe2+ at 25 degrees C; partial reconstitution occurred when 2-mercapto-ethanol was substituted for dithiothreitol. Cortical tryptophan hydroxylase acutely inactivated by methamphetamine or p-chloroamphetamine could be similarly reactivated. MDMA-inactivated cortical tryptophan hydroxylase derived from rats killed later than 3 days after drug treatment could not be significantly reactivated under the conditions described above, indicating the development of irreversible enzymatic damage. Kinetic analysis of enzyme reactivation revealed an approximate doubling of enzyme Vmax with no change in enzyme affinity for either substrate, tryptophan, or pterin cofactor. These studies suggest that MDMA and its congeners inactivate central tryptophan hydroxylase by inducing oxidation of key enzyme sulfhydryl groups. The reactivation capacity of drug-inactivated enzyme at various times after MDMA treatment may provide a means of assessing the development of MDMA-induced neurotoxicity. PMID- 2568405 TI - Effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine in the dog: effect of pargyline pretreatment. AB - Adult beagle dogs of either sex were injected with 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP)-HCl (2.5 mg/kg, i.v.) alone or after pretreatment with pargyline (5.0 mg/kg, s.c., twice), with pargyline alone, or were uninjected. Groups were killed 2 h, 3 weeks, or 3 months after injection, and several brain areas were assayed for biogenic amines and their synthetic and degradative enzymes. MPTP caused a massive and permanent loss of striatal dopamine, tyrosine hydroxylase, and 3,4-dihydroxyphenylalanine decarboxylase activities and the loss of cells within the substantia nigra pars compacta. Dopamine and norepinephrine also were depleted to various degrees in cortex, olfactory bulb, and hypothalamus; however, dopamine beta-hydroxylase activity in cortex was normal. There was no cell loss in the ventral tegmental area or locus ceruleus. The activities of monoamine oxidase (MAO)-A and MAO-B in cortex and caudate were not affected by MPTP. Despite a permanent loss of the nigrostriatal system, the dogs exhibited only a transient hypokinesia lasting 1-2 weeks. Pargyline pretreatment prevented the loss of striatal dopamine and cells from the substantia nigra, but did not prevent a prolonged but reversible decrease in the concentration of dopamine metabolites. It is argued that this apparent inhibition of MAO is due not to suicide inactivation of the enzyme by MPTP, but to reversible inhibition by accumulation of the pyridinium metabolite, 1-methyl-4-phenylpyridinium, selectivity in aminergic terminals. PMID- 2568406 TI - Ionic composition of microdialysis perfusing solution alters the pharmacological responsiveness and basal outflow of striatal dopamine. AB - While using the technique of in vivo microdialysis, we have assessed the effect of the ionic composition of the perfusing solution on extracellular dopamine levels during resting conditions and following a pharmacological manipulation. Our results indicate that perfusion with solutions containing the ionic composition of commercially available Ringer's solution, which mimic the ionic composition of plasma as opposed to brain extracellular fluid, alters the turnover rate and basal release of dopamine. Moreover, perfusion with solutions containing higher calcium levels, i.e., 3.4 mM, than the amount we have determined to be present in the extracellular fluid of striatum (1.2 mM) alters the pharmacological responsiveness of the nigrostriatal dopamine system to synthesis inhibition. PMID- 2568407 TI - Stimulation-induced release of coexistent transmitters in the prefrontal cortex: an in vivo microdialysis study of dopamine and neurotensin release. AB - Extracellular fluid levels of dopamine and neurotensin in the rat prefrontal cortex were measured using in vivo microdialysis. Electrical stimulation of the median forebrain bundle resulted in increased release of both dopamine and neurotensin from the prefrontal cortex. Thus, stimulation of neurons in which dopamine and neurotensin are colocalized can evoke the in vivo release of both substances. PMID- 2568408 TI - Regulation of subtypes of beta-adrenergic receptors in rat brain following treatment with 6-hydroxydopamine. AB - The technique of quantitative autoradiography has been used to localize changes in the densities of subtypes of beta-adrenergic receptors in rat brain following treatment with 6-hydroxydopamine. Previously reported increases in the density of beta 1-adrenergic receptors in the cerebral cortex were confirmed. The anatomical resolution of autoradiography made it possible to detect changes in the density of beta 2-adrenergic receptors in the cortex and in a number of other brain regions. The density of beta 1-adrenergic receptors increased from 30 to 50% depending on the region of the cortex being examined. The increase in the somatomotor cortex was greater than that in the frontal or occipital cortex. The increase in the density of beta 2-adrenergic receptors in the cortex was not as widespread as that of beta 1-adrenergic receptors and occurred primarily in frontal cortex, where the density of receptors increased by 40%. The densities of both beta 1- and beta 2-adrenergic receptors increased in a number of forebrain, thalamic, and midbrain structures. Selective changes in the density of beta 1 adrenergic receptors were observed in the superficial gray layer of the superior colliculus and in the amygdala. The density of beta 2-adrenergic receptors increased in the caudate-putamen, the substantia nigra, and the lateral and central nuclei of the thalamus, whereas the density of beta 1-adrenergic receptors did not change in these regions. The densities of both subtypes of beta adrenergic receptors increased in the hippocampus, the cerebellum, the lateral posterior nucleus of the thalamus, and the dorsal lateral geniculate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568409 TI - The location and function of NMDA receptors in cat and kitten visual cortex. AB - The role of N-methyl-D-aspartate (NMDA) receptors in cat visual cortex was studied as a function of both layer and age by iontophoresis of the NMDA antagonist (D)-2-amino-5-phosphonovaleric acid (APV). Effects on both visual responses and spontaneous activity were observed. In superficial layers (II and III), D-APV reduced visual responses substantially at all ages. Iontophoresis of D-APV with 10 nA of ejecting current for 2-3 min was sufficient to reduce the response to approximately one third of control levels. The magnitude of the reduction did not vary with age. In granular and deep layers (IV, V, and VI), D APV affected the visual response in young animals but only spontaneous activity in older animals. On average, visual responses were reduced to about half at 20 23 days of age and to about 75% at 4 weeks of age but in most cases were not significantly affected in adults. The rapid change in the functional effect of NMDA receptors over the third and fourth week in granular and deep layers suggests a role in development. There was a reasonable age correlation between the change in effect and the period of geniculocortical afferent segregation. Further experiments will be necessary to determine whether NMDA receptors are necessary for segregation to occur. The presence of an NMDA component to the visual response in the adult in layers II and III argues either that these layers retain some form of plasticity in the adult or that NMDA receptors play a role in the transmission of normal visual input to these layers. PMID- 2568410 TI - Contact-mediated loss of the nonsynaptic response to transmitter during reinnervation of an identified leech neuron in culture. AB - We have examined the modification of responses to transmitter during the reformation of a specific synapse by identified leech neurons in culture. Single pressure-sensitive mechanosensory (P) neurons in culture and the soma of this cell in vivo have 2 conductances that are activated by application of 5-HT: a Cl conductance and a monovalent cation conductance (gCations); synaptic release of 5 HT by a serotoninergic Retzius cell in vivo and in culture activates only the Cl conductance and not gCations in the P cell. We have characterized the loss of gCations in P cells by manipulating the culture conditions. When 5-HT was applied from a pipette, innervated P cells and P cells paired with Retzius cells that had not formed synapses had a gCations that was markedly reduced compared with gCations in the single P cell. When 5-HT was included in the culture medium in which single P cells were grown, gCations was not reduced. When Retzius-P cell pairs were treated with reserpine, which was shown to deplete the presynaptic neuron of 5-HT, gCations in the P cell was as low as in untreated pairs. Pairing aldehyde-fixed Retzius cells with untreated P cells also resulted in the loss of gCations. The Retzius cell had both types of receptors, but synapse formation did not affect gCations in the presynaptic neuron. The results demonstrate that the loss of the nonsynaptic response of the P cell to transmitter is due to contact between the neurons rather than to the release of transmitter or trophic factors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568412 TI - Ultrastructural basis for interactions between central opioids and catecholamines. II. Nuclei of the solitary tracts. AB - Interactions between central opioids and catecholamines are thought to underlie the ability of adrenergic agonists both to lower blood pressure and alleviate certain symptoms of opiate withdrawal. We examined the cellular substrate for interactions between neurons containing enkephalin-like opioid peptides and catecholamines in cardiovascular portions of the medial nuclei of the solitary tracts (m-NTS) of adult rats. Single sections were dually labeled using a double bridged peroxidase method for the localization of a monoclonal leucine (Leu5) enkephalin-antibody and immunoautoradiography for the localization of polyclonal antibodies against the catecholamine-synthesizing enzyme tyrosine hydroxylase (TH). Light microscopy revealed a few perikarya and numerous varicosities containing Leu5-enkephalin-like immunoreactivity (LE-LI). These were distributed among TH-labeled perikarya and processes throughout the rostrocaudal NTS. Electron microscopy of the m-NTS at the level of the area postrema further established the single as well as dual localization of TH and LE-LI in individual perikarya, dendrites, and axon terminals. Silver grains indicative of TH-labeling were usually distributed throughout the cytoplasm, whereas the peroxidase reaction product for LE-LI was localized principally to large (80-150 nm), dense core vesicles. Immunoautoradiographic labeling for TH was detected in 118 terminals within a series of sections containing 183 terminals with LE-LI. Of these, 26% of the TH-labeled terminals and 32% of the enkephalin-containing terminals formed symmetric synapses with unlabeled dendrites, while only 7% of each type formed symmetric synapses with TH-labeled dendrites. In favorable planes of sections, the unlabeled as well as TH-labeled dendrites received convergent input from both types of terminals. A few of the remaining terminals that contained either TH or LE-LI formed asymmetric junctions with unlabeled distal dendrites; the others were without recognizable synaptic specializations within the plane of section. Approximately 20% of the TH-labeled terminals and 6% of the terminals containing LE-LI were dually labeled for both antibodies. These were invested with astrocytic processes characterized by bundles of intermediate filaments. We conclude that within cardiovascular portions of the m-NTS, opioid peptides and catecholamines contained within the same or separate terminals modulate the activity of target neurons through direct symmetric, probably inhibitory, synaptic junctions and may additionally modulate the activity of neighboring astrocytes through exocytotic release from large dense-core vesicles.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2568411 TI - Beta 1- and beta 2-adrenergic 125I-pindolol binding sites in the interpeduncular nucleus of the rat: normal distribution and the effects of deafferentation. AB - The plasticity of the beta 1- and beta 2-adrenergic receptor subtypes was examined in the interpeduncular nucleus (IPN) of the adult rat. The beta adrenergic receptor antagonist 125I-pindolol (125I-PIN) was used in conjunction with the selective subtype antagonists ICI 118,551 and ICI 89,406 to determine the subnuclear distribution of beta 1- and beta 2-adrenergic receptors in this nucleus and to correlate the receptor distribution with the distribution of both noradrenergic afferents from the locus coeruleus (LC) and non-noradrenergic afferents from the fasiculus retroflexus (FR). The density of these binding sites was examined following lesions that decreased (LC lesions) or increased (FR lesions) the density of the noradrenergic projection in the IPN. Quantitative radioautography indicated that beta 1-labeled binding sites account for the larger percentage of binding sites in the IPN. The beta 1-binding sites are densest in the those subnuclei that receive a noradrenergic projection from the LC: the central, rostral, and intermediate subnuclei. beta 1-binding sites are algo homogeneously distributed throughout the lateral subnuclei, where there is no detectable noradrenergic innervation. beta 2-binding sites have a more restricted distribution. They are concentrated in the ventral half of the lateral subnuclei, where they account for 70% of total 125I-PIN binding sites. beta 2 binding sites are also present along the ventral border of the IPN. Some of this labeling extends into the central and intermediate subnuclei. Bilateral lesions of the LC, which selectively remove noradrenergic innervation to the IPN, result in an increase in the beta 1-binding sites. Bilateral lesions of the FR, which remove the major cholinergic and peptidergic input from the IPN, elicit an increase in noradrenergic projections and a decrease in beta 1-binding sites. beta 1-binding sites thus exhibit both up-regulation and down-regulation which is correlated with the density of the noradrenergic projection. Our results suggest, therefore, that the density of beta 1-binding sites is regulated by noradrenergic input. beta 2-binding sites increase in density in response to both the LC and FR lesions, suggesting that they are postsynaptic to both of these afferents. The distribution suggests that some of these binding sites may reflect binding to glial cells. The beta 2-binding sites may therefore be regulated by both noradrenergic and non-noradrenergic mechanisms. PMID- 2568413 TI - The role of folate and vitamin B-12 in neurotransmitter metabolism and degenerative neurological changes associated with aging: proceedings of a workshop. AB - Under the sponsorship of the National Institute on Aging and the National Institute of Diabetes and Digestive and Kidney Diseases, the Life Sciences Research Office of the Federation of American Societies for Experimental Biology held a workshop entitled, "The Role of Folate and Vitamin B-12 in Neurotransmitter Metabolism and Degenerative Neurological Changes Associated with Aging." The purpose of the May 1988 workshop was to bring together scientists from various disciplines to identify opportunities for research on an important topic relating to neuroscience, nutrition and aging. PMID- 2568414 TI - Atrial natriuretic peptide and sodium azide dependent guanylate cyclase activities in placentas from normal and severely toxemic patients. AB - Particulate guanylate cyclase is stimulated by several hormones through receptor dependent and by nitrosovasodilators through receptor-independent mechanisms. A subtype of atrial natriuretic peptide (ANP) receptors is coupled to guanylate cyclase. It has been shown that there is a down-regulation of the affinity of ANP receptors to alpha-hANP in placental plasma membranes obtained from severely toxemic patients. We have asked the question whether these changes are associated with a down-regulation of ANP-dependent guanylate cyclase activity. Guanylate cyclase was determined by in vitro experiments using a placental plasma membrane fraction obtained from normal and from severely toxemic patients. The presence of ANP-dependent placental guanylate cyclase activity was demonstrated both in normal and toxemic placentas. Although basal guanylate cyclase activity was not influenced by toxemia of pregnancy, there was a significant decrease in the maximum stimulation of this enzyme by alpha-hANP (104.81 +/- 12.02% (n = 4) vs 49.41 +/- 8.73% (n = 7) for normal and toxemics, respectively). Finally, stimulation by a nitrosovasodilator, sodium azide (NaN3), was also lower in toxemic placentas than in normal controls. These observations extend our previously reported results on placental ANP receptor function but also suggest the presence of a possibly receptor-independent decrease in guanylate cyclase activity in toxemic placentas. PMID- 2568415 TI - Histologic evaluation of bone inductive proteins complexed with coralline hydroxylapatite in an extraskeletal site of the rat. AB - The purpose of this histologic and biochemical study was to assess the osteogenic potential of bone inductive proteins complexed with coralline hydroxylapatite as the carrier vehicle after implantation in an extraskeletal site of the rat. Inductive proteins were extracted from bovine demineralized bone. Implants were placed in 16 male, 3-month old Long-Evans rats (200-300 grams), using paired subcutaneous sites overlying the ventral thorax. There were four experimental groups, with eight implants per group. These included hydroxylapatite alone (HA), hydroxylapatite with inductive protein (HA + P), inactive demineralized bone matrix with (IBM + P), and without inductive protein (IBM). All implants were harvested at 21 days. Findings indicate a lack of osteogenic potential in groups HA, HA + P, and IBM. However, when HA and HA + P were compared, there was a 79% increase in standardized field mean nuclear point counts in the HA + P group. Also, compared to the other three implant groups, controls of IBM + P histomorphometrically showed chondroid bone formation and increased alkaline phosphatase activity. In this model system it may be concluded that with a composite system of coralline hydroxylapatite and bovine-derived inductive protein, bone formation was not seen; positive controls consisting of IBM + P demonstrated a statistically significant increase in AP activity with corresponding histologic evidence of bone formation. PMID- 2568416 TI - The antiemetic profile of zacopride. AB - The antiemetic activity of zacopride against a variety of emetogenic agents has been determined in dogs. Zacopride was highly effective in inhibiting emesis due to a wide range of cancer chemotherapeutic agents, particularly cisplatin. It was well absorbed orally since the dose of zacopride required to inhibit cisplatin induced emesis in dogs by 90% was 28 micrograms kg-1 both by i.v. and p.o. routes. Further, zacopride (1 mg kg-1 p.o.), administered after the onset of cisplatin-induced emesis, reduced the number of subsequent emetic episodes by 91%. Zacopride at 0.1, 1, or 3.16 mg kg-1 p.o. or i.v., reduced the number of emetic episodes due to dacarbazine, mechlorethamine, adriamycin, actinomycin D, or peptide YY by 100, 100, 86, 96 and 79%, respectively. However, zacopride was not effective in inhibiting emesis due to either apomorphine, copper sulphate, protoveratrine A, histamine, or pilocarpine. No adverse effects attributed to zacopride were observed. Zacopride is thus a unique and potent antiemetic agent as it selectively inhibits the emetic response to cancer chemotherapy agents and peptide YY. PMID- 2568418 TI - Effects of anti-inflammatory drugs on interleukin 1-induced cartilage proteoglycan resorption in-vitro: inhibition by aurothiophosphines but no influence from perturbed eicosanoid metabolism. AB - A range of anti-inflammatory drugs having varying effects on eicosanoid metabolism and other actions was studied for their potential to inhibit alpha interleukin 1 (IL-1)-induced cartilage proteoglycan resorption in-vitro. No significant effects on resorption were observed with inhibitors of cyclo oxygenase, lipoxygenase or mixed inhibitors of both these enzymes, and no influence on IL-1 effects was observed with added eicosanoids. Among the clinically used disease modifying anti-arthritic agents, only auranofin and the immunoregulatory agent, tilomisole, were found effective in inhibiting resorption. Some auranofin analogues having chloride or nitrate leaving groups that inhibit DNA polymerase-alpha were found to be potent inhibitors of IL-1 induced resorption. PMID- 2568417 TI - Definition of the in-vivo binding of [3H]spiperone in rat brain using substituted benzamide drugs. AB - The ability of some substituted benzamide drugs to define in-vivo the binding of [3H]spiperone to brain dopamine receptors in rats was assessed using behaviourally effective doses in comparison with haloperidol. As judged using haloperidol, [3H]spiperone identified dopamine receptors in the substantia nigra, striatum, tuberculum olfactorium and hypothalamus, but not in frontal cortex or nucleus accumbens. The substituted benzamide compounds alizapride, metoclopramide, clebopride and YM 09151-2 prevented the accumulation of [3H]spiperone in the substantia nigra, striatum, tuberculum olfactorium and hypothalamus. However, YM 09151-2 also caused displacement of [3H]spiperone accumulation in the nucleus accumbens and frontal cortex. (+/- )-Sulpiride, (+/- )-sultopride, amisulpiride and prosulpride all prevented the accumulation of [3H]spiperone in the hypothalamus but were ineffective in one or more of the other regions containing dopamine receptors defined by [3H]spiperone. The isomers of sulpiride and sultopride stereoselectively defined the accumulation of [3H]spiperone in dopamine containing brain regions. The (-)-isomers of both drugs prevented the accumulation of [3H]spiperone in the substantia nigra, striatum, tuberculum olfactorium and hypothalamus. In contrast, (+)-sulpiride and (+) sultopride were ineffective. Selected substituted benzamide drugs can be used to define the interaction of ligands with dopamine receptors in-vivo. These substances may be useful in PET studies in man. The isomers of some substituted benzamine drugs may be used to define dopamine receptors in-vivo by enantiomeric selectivity. PMID- 2568419 TI - Topical application of penetration enhancers to the skin of nude mice: a histopathological study. AB - Eighteen potential penetration enhancers, some at concentrations that might be used for that purpose, have been examined to evaluate their irritancy potential on nude mouse skin. A biopsy technique was employed followed by histological examination. Up to 50% glycerol, 10% hydroxyethyl lactamide (HELA), 10% oleyl alcohol, 10% Solketal, 10% glycofurol, 100% tetrahydrofurfuryl alcohol (THFA) and 10% urea induced no discernible change in the histological appearance of the skin whereas 100% dimethyl sulphoxide (DMSO), 100% dimethyl formamide (DMF), 100% N methyl-2-pyrrolidone, 10% Azone, 10% oleic acid, 10% methyl laurate, 10% benzyl alcohol and 10% glycerol formal caused severe skin irritation. PMID- 2568420 TI - Thermodynamics of the binding of salicylate to human serum albumin: evidence of non-competition with imidazole. AB - The thermodynamic characteristics of the binding of salicylate to human serum albumin have been studied using a technique based on the variation of the quantum yield of fluorescence of salicylate when it binds to the protein. The binding constants, number of sites and the values of delta G degrees, delta H degrees and delta S degrees were determined. The results are consistent with a model that proposes two equal and independent types of binding site with a predominantly ionic interaction and an important hydrophobic contribution in one of the sites. The technique was also used to demonstrate that imidazole and salicylate (that can be found simultaneously in plasma following administration of imidazole-2 hydroxybenzoate) do not compete for the same binding sites on the protein. PMID- 2568421 TI - Enhancement of phenytoin binding to tissues in rats by heat treatment. AB - Phenytoin binding to heat-treated tissue homogenates has been examined to characterize the phenytoin binding to tissues. The binding to the heat-treated tissue homogenates was enhanced in all tissues studied compared with controls. The heating might produce the changes in conformation of proteins in tissues and then enhance phenytoin binding to tissue homogenates. PMID- 2568422 TI - Effect of some penicillins on the sensitivity of limulus amoebocyte lysate test. AB - A group of penicillins have been tested for their effect on the sensitivity of limulus amebocyte lysate test (LAL). Cloxacillin, cephalothin and cefuroxime inhibited the gel formation at concentrations above 2 mg mL-1, while ampicillin, methicillin and mecillinam showed no inhibitory effects upto concentrations of 10 mg mL-1. For benzylepenicillin the maximum non-inhibitory concentration was 10,000 units mL-1. However, the dilutions required to overcome inhibition were within the limits of maximum valid dilutions computed for each product showing that LAL test is valid for these products at their non-inhibitory concentrations. PMID- 2568423 TI - The influence of structure on the accumulation of caffeine induced by methyl xanthine derivatives. AB - In rats given caffeine (25 mg kg-1 p.o.) and 1,3,8-trisubstituted xanthine (1,3,8 TSX) derivatives (10 mg kg-1 p.o.) the accumulation of the former in plasma was 300% higher than that in control animals given caffeine alone. The effect on caffeine accumulation appears to be independent of the nature of the N3 substituent and its absence in rats given 1,3-disubstituted xanthines (1,3-DSX) instead of 1,3,8-TSX suggests that the presence of the C8-methyl group in the latter compounds is responsible for the accumulation phenomenon. The results of our previous work imply that these observations in this rat model can be extrapolated to man. PMID- 2568424 TI - Interaction between the cardiovascular effects of clonidine and the kappa-opioid agonist U-50,488H in the anterior hypothalamic area of the rat brain. AB - In thiobutabarbitone-anaesthetized rats, microinjection of clonidine (1-40 nmol) into the anterior hypothalamic area (AHy) produced dose-dependent reductions in mean arterial blood pressure and heart rate. Microinjection of the kappa-opioid agonist U-50,488H (3 and 10 nmol) did not modify these parameters. Simultaneous co-administration of clonidine (4 nmol) and U-50,488H (10 nmol) into the AHy resulted in significant potentiation of the clonidine-induced hypotension and marked attenuation of the bradycardia. A lower dose of U-50,488H (3 nmol) co administered with clonidine (4 nmol) did not influence the cardiovascular responses to clonidine. These findings suggest that AHy neurons involved in the cardiovascular responses to clonidine may be modulated by kappa-opioid receptor stimulation. PMID- 2568425 TI - Distribution of dissociation constant values of muscarinic agonists. AB - The frequency distributions of dissociation constant values of some muscarinic agonists (carbachol, muscarone and cis-2-methyl-5-trimethylammoniummethyl-1,3 oxathiolane) obtained on guinea-pig ileum and atria and rat urinary bladder have been examined to see if the means of the dissociation constant values and the statistical tests for their significance, should be based on geometric rather than on arithmetic means. For the three compounds the distributions on a logarithmic scale did not significantly deviate from normality while the distributions on an arithmetic scale tended to deviate from normality. PMID- 2568426 TI - Determination of cephalosporin-C amidohydrolase activity with fluorescamine. AB - A spectrophotometric procedure for the assay of cephalosporin-C amidohydrolase activity, based on the determination of the 7-aminocephalosporanic acid (7-ACA) produced in the hydrolysis of cephalosporin-C by the enzyme, is described. This procedure can be used to detect 7-ACA over a range of 10 to 200 micrograms mL-1. The same method can be used as a fluorometric procedures with a 100-fold greater sensitivity. At pH 4.5 7-ACA produces a strong fluorophor with fluorescamine, detectable spectrophotometrically at 378 nm and fluorometrically at an excitation of 378 nm and emission of 495 nm. At this pH the fluorophors formed with cephalosporin-C, proteins and aminoadipic acid present minimal absorbance values. The conditions for maximal detection of 7-ACA in the presence of proteins, cephalosporin-C and aminoadipic acid have been determined. PMID- 2568427 TI - Effects of 3,3'-di-O-methylquercetin on guinea-pig isolated smooth muscle. AB - The effects of the flavone 3,3'-di-O-methylquercetin (DOMQ) have been examined and compared with those of quercetin, on guinea-pig isolated ileum, trachea, and main pulmonary artery (MPA). Except for transient contractions induced by low concentrations (10(-8)-3 x 10(-6) M), DOMQ and quercetin (up to 3 x 10(-4) M) caused reduction of the tone and the phasic contractions of the ileum. A23187 reversed the inhibitory effects of quercetin but not those of DOMQ. DOMQ and quercetin caused concentration-dependent relaxation of the trachea and the adrenaline-contracted MPA. DOMQ shifted to the right the concentration-effect curves induced by acetylcholine on the ileum and the trachea, and by adrenaline on MPA and those induced by CaCl2 on ileum, trachea and MPA. DOMQ also inhibited the contractions induced, in Ca2+-free EGTA-containing buffer, by histamine on ileum and by adrenaline on MPA. These observations suggest that DOMQ inhibits Ca2+ influx, Ca2+ release from intracellular stores and, more likely, Ca2+ binding to intracellular receptor proteins. PMID- 2568428 TI - Ordered powder mixtures: reality or fiction? PMID- 2568429 TI - Comments to 'order out of chaos'. PMID- 2568430 TI - Chronic but not acute antidepressant treatment increases pentetrazol-induced convulsions in mice. PMID- 2568431 TI - The in-vitro pH-dissolution dependence and in-vivo bioavailability of frusemide PVP solid dispersions. AB - The dependence of the dissolution rate on the pH of the buffered medium, using constant surface area discs, has been examined for crystalline frusemide, a semi crystalline frusemide-polyvinylpyrrolidone (PVP) solid dispersion and an X-ray amorphous frusemide-PVP dispersion. The marked changes observed in the pH dissolution profiles indicate that differing dissolution mechanisms operate in the amorphous regions. This conclusion was further supported by the comparison of pH-dissolution and pH-equilibrium solubility profiles that suggested a supersaturation effect to be the relevant term in describing the dissolution enhancing effects of amorphous regions. A marked dissolution enhancement, relative to crystalline frusemide, was shown by the X-ray amorphous solid dispersion in weakly acidic solutions. A similar effect was observed in the dissolution characteristics of gelatin capsule formulations in simulated gastric and intestinal media. In a human bioavailability study, the X-ray amorphous frusemide-PVP solid dispersion exhibited a significant reduction in the time for maximum effect in comparison to crystalline frusemide and a semi-crystalline solid dispersion. This effect, demonstrated by the primary end organ response in seven healthy subjects, concurred with the in-vitro prediction of dissolution enhancement in weakly acidic media. PMID- 2568432 TI - Methodology for a better evaluation of the relation between mechanical strength of solids and polymorphic form. AB - In order to evaluate the role played by polymorphism in the mechanical strength of solid dosage forms (e.g. compressed tablets) and minimize the influence of other factors (such as compaction force, porosity, particle size, and possibly crystal habit), a melted disc technology was developed. With this technique, tablet-shaped discs of zero porosity were prepared by melting powder and subsequent crystallization in the desired modifications. Taking phenobarbitone as a model drug, different methods were used to get discs of forms I, II and III and the amorphous form. Mechanical properties of the discs were assessed, primarily through their bending strength. The Vickers hardness number was also determined for some specimen discs and monocrystals. Results showed that the amorphous form and form III of phenobarbitone gave the toughest discs and would therefore the most suitable materials to manufacture coherent tablets. Moreover, the various preparation methods used resulted in discs of different internal structures. Both crystal size and crystal habit significantly affected the physical properties of the tested materials. PMID- 2568433 TI - Solubilization of indomethacin by polysorbate 80 in mixed water-sorbitol solvents. AB - The effect of indomethacin on the micellar properties of the non-ionic surfactant, polysorbate 80, in water-sorbitol mixtures containing up to 25% w/v sorbitol has been investigated by light scattering, photon correlation spectroscopy and viscometric techniques. The molecular weight of polysorbate 80 micelles containing solubilized indomethacin increased linearly with increase of sorbitol concentration. Solubilization of indomethacin resulted in an increase of micellar weight due not only to the incorporation of solubilizate but also to an increase in the number of polysorbate molecules per micelle. The micelles in all systems were most satisfactorily represented as oblate ellipsoids, the asymmetry and hydration of which increased with increase of sorbitol concentration. Indomethacin solubilization caused a restructuring of the micelle to produce a more symmetrical micelle of increased hydration. PMID- 2568434 TI - Visceral leishmaniasis: drug carrier system characteristics and the ability to clear parasites from the liver, spleen and bone marrow in Leishmania donovani infected BALB/c mice. AB - The efficacy of various sodium stibogluconate formulations against Leishmania donovani has been investigated using a BALB/c mouse model of visceral leishmaniasis. Only one therapy, multiple dosing with drug loaded sonicated vesicles, liposomes or niosomes, was found to be effective against parasites in the liver, spleen and bone marrow. Other treatments significantly reduced parasite liver burdens but either failed to effect spleen and bone marrow parasites, or were effective but toxic. Prophylactic treatment with sodium stibogluconate preparations, six days before infection, reduced parasite multiplication in the liver (free, niosomal and liposomal drug) and the spleen (sonicated, drug loaded niosomes only), but had no suppressive effect on bone marrow parasite burdens compared with controls. These results indicate that in vivo sodium stibogluconate persists in some compartments at parasiticidal concentrations and that failure to reach this concentration at some sites of infection such as bone marrow, is the cause of treatment failure and relapse. PMID- 2568436 TI - The enhanced daily excretion of urinary methylamine in rats treated with semicarbazide or hydralazine may be related to the inhibition of semicarbazide sensitive amine oxidase activities. AB - The effects of amine oxidase inhibitors upon the daily urinary excretion of monomethylamine (MMA), dimethylamine (DMA), trimethylamine (TMA) and ammonia in the rat have been examined. Administration of hydralazine (5 mg kg-1) or semicarbazide (100 mg kg-1), drugs which irreversibly inhibit semicarbazide sensitive amine oxidases (SSAO) but not monoamine oxidase (MAO), enhanced MMA excretion by around three- to six-fold above pretreatment levels, whereas no effect of pargyline (25 mg kg-1), a selective irreversible inhibitor of MAO was found. No apparent changes in DMA or TMA excretion in response to drug-treatment were observed. Ammonia excretion also was generally unchanged except for an apparent marked increase (approximately four-fold) over the 24 h following semicarbazide, a result which might be explained if ammonia is a degradation product of semicarbazide metabolism in the rat. With recent evidence that MMA is a substrate in-vitro for SSAO activities, results here may indicate that SSAO or related enzymes are involved in endogenous MMA turnover. PMID- 2568435 TI - Lack of mixed agonist-antagonist properties of [Gln8-Gly31]-beta h-EP-Gly-Gly-NH2 and [Arg9,19,24,28,29]-beta h-EP in the rat vas deferens neuroeffector junction: studies with naloxone, beta-funaltrexamine and ICI 174,864. AB - The 1-27 truncated fragment of beta h-endorphin (beta h-EP) as well as [Gln8,Gly31]-beta h-EP-Gly-Gly-NH2 or [Arg9,19,24,28,29]-beta h-EP exhibited opiate agonist activity in the rat vas deferens bioassay; the potency of these peptides was 3 to 6 times less than that of beta h-EP. None of these compounds exhibited any degree of antagonism towards the inhibitory action of beta h-EP. Naloxone antagonized and reversed the inhibitory action of beta h-EP and its analogues though with varying potencies. The apparent naloxone-pA2 value for beta h-EP was 8.94; that for [Gln8-Gly31]-beta h-EP-Gly-Gly-NH2 was 8.08 and that for [Arg9,19,24,28,29]-beta h-EP was 8.38. beta-Funaltrexamine (beta-FNA) potently antagonized the inhibitory action of beta h-EP following non-equilibrium kinetics. Tissue preincubation with 10 nM beta-FNA for 60 min followed by extensive washing caused a 10-fold increase in the beta h-EP IC50. However, 10 nM beta-FNA caused only a 1.2 increase in the IC50 of [Gln8,Gly31]-beta h-EP-Gly-Gly NH2 and a 4.1-fold increase in the IC50 of [Arg9,19,24,28,29]-beta h-EP. In contrast, preincubation of the tissue with 3 microM ICI 174,864 did not modify the potency of beta h-EP or its structural analogues. However, a 60 min pretreatment with 10 microM beta-FNA followed by the addition of 3 microM ICI 174,864 revealed a further decrease in the potency of the opiopeptins compared with tissues exposed to beta-FNA alone or ICI 174,864 alone. In conclusion, the inhibitory action of these peptides is remarkably sensitive to beta-FNA antagonism; in addition the peptides act as pure opiate agonists in marked contrast with the agonist-antagonist properties described in the CNS. PMID- 2568438 TI - The effect of machine speed on the consolidation of four directly compressible tablet diluents. AB - The reduction in porosity of a powder bed on compression was found to be a function of the velocity of the punch of the press. Substances which consolidated principally by fragmentation showed relatively little velocity dependence. However, the more important deformation was in a powder's consolidation mechanism, the greater the dependence on punch velocity and hence tablet press speed. PMID- 2568437 TI - Solid state adsorption of antibiotics onto sorbitol. AB - The ability of two types of sorbitol, instant and crystalline, to hold antibiotics permanently after mixing has been assessed by an air sieving technique. Sorbitol instant was found to have a greater adsorption capacity and binding strength than crystalline sorbitol. The six antibiotics studied were found to fall roughly into two groups of different adsorption capacities: (1) pivampicillin, cephalexin monohydrate and erythromycin ethylsuccinate, and (2) ampicillin trihydrate, amoxycillin trihydrate and cloxacillin sodium. The former have slightly higher levels of adsorption than the latter. A negative linear relationship was found between the amount of antibiotic adsorbed onto dry sorbitol and that originally added to sorbitol. When adsorption is expressed as the weight of drug adsorbed per unit weight of sorbitol, an 'apparent' Langmuir isotherm results. This suggests that there are a number of adsorption sites available for holding drug particles, these sites being different for the different antibiotics. PMID- 2568439 TI - Hydrogel patches for transdermal drug delivery; in-vivo water exchange and skin compatibility. AB - Hydrogel patches based on water swellable polyacrylates have been developed for long-term transdermal drug delivery. Two properties, relevant to the performance of hydrogel patches in-vivo have been investigated in humans over five days. These were: (i) the kinetics of water exchange between the skin and the patches; (ii) the skin compatibility of the patches. It was found that initially there was a gradually increasing uptake of water from the skin by the patches, but after about 20 h the water exchange followed a regular fluctuating pattern, peaking once a day and once a night. The skin compatibility of the patches was satisfactory, in that no redness or pustulation was noticed throughout the five days. This was most likely due to the capability of the patches to exchange water with the skin. PMID- 2568440 TI - The effect of some preservatives used in nasal preparations on mucociliary clearance. AB - The effect of methyl-p-hydroxybenzoate, propyl-p-hydroxybenzoate, chlorbutol, chlorocresol, EDTA, benzalkonium chloride, chlorhexidine, phenylmercuric nitrate and phenylmercuric borate on mucociliary transport rate of the frog palate has been examined. Following a variable number of applications all these preservatives halted transport, the first three reversibly. However, applications of thiomersal (0.01%) were well tolerated. The frog palate possesses a ciliated epithelium protected by mucus, since some of our findings are at variance with those previously reported results where the protective effect of mucus was negligible in the in-vitro model (usually trachea) employed, it would appear that the contribution of mucus to effective mucociliary clearance should not be underestimated. PMID- 2568441 TI - Pharmacokinetics of quinacrine after intrapleural instillation in rabbits and man. AB - Quinacrine was given by intrapleural instillation or intravenous infusion to 10 rabbits. The uptake of quinacrine from the pleural space was rapid and complete. The mean absorption half-life was approximately 7 min and the mean bioavailability was slightly in excess of 100%. Similar absorption characteristics generally applied in man, in a pilot study on four patients. In three of them, peak quinacrine plasma concentrations were reached that were far above the normal therapeutic range. Known systemic side-effects of quinacrine comprise CNS stimulation, toxic psychosis and convulsions. In view of the high bioavailability and the large doses used for pleural sclerosing (pleurodesis) in patients, neurological disease and psychiatric disturbances that predispose to CNS toxicity should be considered as contraindications to intrapleural quinacrine. PMID- 2568442 TI - Phorbol esters of different biological activities may preferentially act as mitogens of human suppressor T-cells and are equi-effective mitogens of IL-2 dependent cells. AB - The actions of tetradecanoylphorbolacetate (TPA) and 12-deoxyphorbolphenylacetate 20-acetate (DPPAA) together with a phytohaemagglutinin (PHA) have been examined on the proliferative responses of human mononuclear cells (MNC) depleted of specific cell subsets by the use of monoclonal antibodies. PHA-induced proliferation was found to be reduced when monocytes/macrophages and T-helper cells were depleted from MNC, but enhanced compared with MNC responses when T suppressor cells were depleted. In contrast, TPA- and DPPAA-induced proliferation was unchanged or slightly enhanced following macrophage/monocyte depletion, and whereas TPA-induced proliferation was largely independent of subtype constitution, the non-tumour promoting DPPAA appeared to selectively enhance proliferation of the T8+ suppressor subset. Indomethacin increased the proliferative MNC responses of phorbol esters whilst having little effect upon the PHA response, an effect antagonized by addition of PGE2. The addition of interleukin-2 (IL-2) increased the proliferative response, as well as resistance to inhibition induced by cyclosporin A and dexamethasone and partially abolished the selective actions of DPPAA and PHA. In IL-2 dependent cultures PHA induced stimulation was more sensitive to inhibition by cyclosporin than were the phorbol esters. The results suggest that, although induction of lymphocyte proliferation by phorbol esters is not a correlate for tumour promotion itself, non-promoting phorbol esters may have a more restricted ability to induce proliferation than TPA. PMID- 2568443 TI - Naloxone-insensitive inhibitory and excitatory effects of opioid agonists in the rat isolated uterus. AB - Morphine (3 x 10(-6)-10(-3) M) produced a concentration-dependent inhibition of both spontaneous rhythmic contractions and tonic contraction induced by potassium chloride (KCl) (50 mM) in rat isolated uterus. Uteri at the metoestrus phase were the most sensitive to the inhibitory effect of morphine. Morphine-induced relaxation was characterized by a post-wash tonic contraction. The mu-specific opioid agonist, D-alaglymepheglycol (DAGO) (4 x 10(-8)-1.2 x 10(-5) M) did not affect spontaneous rhythmic contraction but produced a partial inhibition of the KCl depolarized uterus. There was no post-wash contraction following DAGO. In contrast to the inhibition produced by both morphine and DAGO, methionine enkephalin (8 x 10(-7)-2.4 x 10(-5) M) produced a concentration-dependent contraction of the KCl depolarized uterus. Naloxone up to 100 microM, propranolol (10 microM), flurbiprofen (50 microM) and metiamide (10 microM) did not affect either the relaxation produced by morphine and DAGO or the contraction produced by methionine enkephalin. The results showed that the opioid agonists may have non-receptor mediated direct effects on the rat uterus. PMID- 2568444 TI - Effects of urapidil on catecholamine turnover and release in the central nervous system of the rat. AB - The actions of urapidil, prazosin, idazoxan, and haloperidol on the turnover of noradrenaline in the hypothalamus and dopamine in the nucleus accumbens of the rat were investigated using changes in the ratios of 3-methoxy-4-hydroxyphenyl glycol/noradrenaline (MHPG/NA) and 3,4-dihydroxyphenylacetic acid/dopamine (DOPAC/DA), respectively, as measures for drug-induced effects. Urapidil (2.5-30 mg kg-1 i.v.) increased the ratios of MHPG/NA and DOPAC/DA. Its effects on NA turnover were maximal at 60 min (160% of control at 30 mg kg-1), and on DA turnover at 30 min (138% of control at 30 mg kg-1). Prazosin (0.5-2.5 mg kg-1 i.v.) had no effect, but the high dose of 5 mg kg-1 i.v. significantly increased the ratio of MHPG/NA in the hypothalamus. Idazoxan (2-50 mg kg-1 i.v.) and haloperidol (0.02-0.5 mg kg-1 i.v.) selectively enhanced turnover of NA and DA, respectively. In experiments on field-stimulated overflow of tritium from slices of hypothalamus and nucleus accumbens labelled with [3H]NA or [3H]DA, respectively, urapidil (1 mumol L-1) facilitated the evoked responses in both regions. Prazosin (0.1 mumol L-1) had no effect in either of the two areas. Idazoxan (0.1 mumol L-1) increased stimulated overflow of [3H]NA from the hypothalamus but not of [3H]DA from the nucleus accumbens. Conversely, haloperidol (0.1 mumol L-1) greatly enhanced evoked overflow of [3H]DA but not of [3H]NA. From the present results it is concluded that urapidil has an antagonistic effect at central alpha 2-adrenoceptors and also a weak antagonistic action at central dopamine D2-receptors. PMID- 2568445 TI - Non-linear intestinal absorption kinetics of cefadroxil in the rat. AB - Absorption of cefadroxil in a selective intestinal absorption area (the proximal third of the small intestine) of the anaesthetized rat, at seven initial perfusion concentrations, ranging from 0.01 to 10.0 mg mL-1, is shown to be a non linear transport mechanism. With the aid of computer-fitting procedures based on differential and integrated forms of Michaelis-Menten equation, Vm and Km values of 36.7-37.3 mg h-1 and 12.0-13.0 mg, respectively, were found. The statistical parameters were better than those obtained both for first-order and for combined Michaelis-Menten and first-order kinetics. There is no evidence for substantial passive diffusion processes. The results reported here, together with allometric considerations and literature data analysis, may help to explain some particular non-linear features of plasma level curves associated with the administration of fairly high oral doses of cefadroxil to humans. PMID- 2568446 TI - Influence of streptozocin-induced diabetes on reductive metabolism of acetohexamide in rat liver. AB - Streptozocin-induced diabetes significantly decreased acetohexamide reductase activity of 10,000 g supernatant of liver homogenates from both male and female rats. However, the decrease in activity from female rats was smaller than that from male rats, thereby eliminating the sex difference in the activity of the 10,000 g supernatant. In male rats, the diabetes markedly decreased acetohexamide reductase activity only in the microsomal fraction of liver homogenate, whereas in female rats, it decreased the activity only in the cytosolic fraction. These results indicate that the mechanism for the decreasing effect of the diabetes on reductive metabolism of acetohexamide in 10,000 g supernatant differs between male and female rats. PMID- 2568447 TI - The influence of pH on the cardiac depressant action of tobramycin. AB - Tobramycin, like other aminoglycoside antibiotics, is a basic compound which causes a reduction of cardiac contractile force and inhibits the inotropic action of calcium. Its antagonistic action towards calcium had been analysed at different pH values of the solution bathing the rat left atrium. The isometric tension of the electrically driven atrium (4 Hz, 2 ms, supramaximal voltage) in response to cumulative increase in the bath calcium concentration (2.0 to 32.0 mM) was measured in the absence and in the presence of 2.0 x 10(-5) M tobramycin. The increase in pH from 6.6 to 8.2, which decreased the ionization of tobramycin, reduced the cardiac depression. Similar experiments were carried with other cardiac depressants. Thus, the effect of methoxyverapamil (3.5 x 10(-7) M), also a basic compound, was increased at higher values of pH; the acidic pentobarbitone (4 x 10(-5)M) was less effective at more alkaline pH; the cardiac depression of the non-electrolyte ethanol (2 x 10(-4)M) was practically the same over the pH range studied. The time for equilibrium of the antagonism was small for tobramycin, moreover, its antagonism could only be fully reversed by an increase in extracellular calcium. These results suggest a competitive type of antagonism, between the ionized fraction of tobramycin and calcium, for superficial sites of the sarcolemma. The effect of methoxyverapamil, pentobarbitone and ethanol seems to occur at a more internal site of the cardiac cell since the unionized form of the drug is the most active. PMID- 2568448 TI - In-vitro evaluation of filaricidal activity of GABA and 1,3-dipalmitoyl-2-(4 aminobutyryl)glycerol HCl: a diglyceride prodrug. AB - A diglyceride ester of gamma-aminobutyric acid (GABA) has been synthesized and its filaricidal activity compared with GABA, and progabide in-vitro, on infective larvae and microfilariae of Molinema dessetae, a rodent filaria. GABA induced paralysis in infective larvae but was inactive on microfilariae. There were interactions between the culture medium and GABA. The ester drug at 0.1 mmol L-1 (1,3-dipalmitoyl-2-(4-aminobutyryl)glycerol HCl) was as active as progabide on infective larvae and hundredfold more potent than GABA. Its microfilaricidal activity at 1 mmol L-1 was lower than that progabide at 0.1 mmol L-1 but a delayed effect was observed. The data confirm filariae sensitivity to GABA derivatives. PMID- 2568449 TI - The transport of microspheres from the gastro-intestinal tract to inflammatory air pouches in the rat. AB - The distribution of latex microspheres (1.1 microns diam) has been investigated in-vivo, as a potential passive targeted system for the treatment of inflammation. Microspheres administered orally were found in the circulation and in inflamed tissues and exudates of inflammatory air pouches in rats. Oral absorption was also found in a rabbit. Particles administered directly into the circulation also penetrated into the air pouch tissues and fluids. The possibility of using microspheres as a passive targeted system for the treatment of inflammation is discussed. PMID- 2568450 TI - Sustained release ophthalmic formulations of pilocarpine. AB - The bioavailability of drugs from conventional ophthalmic formulations is low. To optimize the therapy, sustained release ophthalmic dosage forms are warranted. Hydrogels such as sodium-carboxymethyl cellulose, hydroxypropylmethyl cellulose, Carbopol-940, Carbopol-941 and Lutrol-FC-127 increase the duration of action of various drugs. Gels containing pilocarpine were prepared and evaluated by measuring the intensity and duration of miotic response in albino rabbits. Carbopol-940 gels, being the best of those used, were studied further for the effect of its concentration and of additives (benzalkonium chloride, phenylmercuric nitrate, chlorbutol and disodium edetate), autoclaving at 121 degrees C for 30 min and irradiation with gamma rays (2.5 Mrad), on the end product. PMID- 2568451 TI - Transport characteristics of [3H]-chlorpromazine across rat small intestinal brush border membrane. AB - The transport mechanism of chlorpromazine, a tertiary amine, has been investigated using brush border membrane vesicles isolated from rat small intestine. Chlorpromazine was taken up rapidly by the vesicles the uptake being mainly due to binding to the membrane. The transport of chlorpromazine into the intravesicular space was facilitated by the transmembrane electrical potential difference (inside negative) induced by valinomycin or sodium thiocyanate. This facilitating effect was observed only when the transmembrane electrical potential difference was induced after chlorpromazine uptake had reached a steady state. In the initial phase of chlorpromazine uptake, there was no effect. Therefore, it is suggested that both rapid binding to brush border membrane and transmembrane electrical potential difference (inside negative) across the membrane plays a significant role in the transport processes of chlorpromazine through the intestinal epithelium. PMID- 2568452 TI - Effect of lithium chloride on the neurotransmitter release from adrenergic nerve terminals of guinea-pig atria. AB - Lithium chloride at 1 and 10 mM produced a dose-dependent inhibition of the cardiac response to field stimulation of the adrenergic nerve terminals, without affecting myocardial contractility in electrically stimulated guinea-pig atria. This effect was calcium-independent and was also present in preparations superfused with 10 mM myoinositol. Moreover, 10 mM lithium chloride reduced the positive inotropic effect of tyramine. These results indicate that lithium reduces the evoked release of noradrenaline from the adrenergic nerve endings, probably lowering the content of releasable neurotransmitter. PMID- 2568453 TI - Deamination of aliphatic amines of different chain lengths by rat liver monoamine oxidase A and B. AB - Monoamines with from 1 to 18 straight chain carbon atoms have been analysed as rat liver monoamine oxidase substrates. Methylamine and ethylamine are clearly not substrates of monoamine oxidase (MAO). n-Propylamine, n-butylamine, n dodecylamine and n-octadecylamine are relatively poor substrates, i.e. with high Km and low Vmax values for the enzyme. n-Pentylamine, n-hexylamine, n heptylamine, n-octylamine, n-nonylamine and n-decylamine are all very good MAO substrates. All these aliphatic amines are found to be typical type B substrates according to the sensitivities of the enzyme towards the selective MAO-B inhibitor selegiline and the MAO-A inhibitor, clorgyline. The sensitivity towards selegiline with respect to these amines is even higher, i.e. Ki = 1 x 10(-9) M for butylamine, than that of the typical type B substrate beta-phenylethylamine (Ki = 1 x 10(-8) M). The sensitivity towards selegiline decreases slightly with increasing chain length of these aliphatic amines. PMID- 2568454 TI - Phenolic analogues of diastereoisomeric 2-methyl reversed esters of pethidine. AB - The preparation and stereochemical characterization of alpha- and beta-isomers of 1,2-dimethyl-4-m-hydroxyphenyl-4-propionyloxypiperidine are described. Both the alpha (axial 4-aryl/chair) and beta (equatorial 4-aryl/chair) isomers were of low potency or inactive in mice antinociceptive tests. Shortcomings of the alpha isomer as a model for the 4-arylpiperidine moiety of morphine are discussed. PMID- 2568455 TI - Comparative effects of ouabain on isolated papillary muscle from tree shrews, guinea-pigs and rats. AB - Inotropic effects of ouabain were investigated in the isolated papillary muscle preparations of the tree shrew (Tupaia glis), guinea-pigs and rats. In the guinea pig and shrew papillary muscles, ouabain at concentrations of 10(-8) to 3 x 10( 7) M caused a concentration-dependent positive inotropic response in a similar magnitude, while in the rat, ouabain at concentrations of 10(-7) to 3 x 10(-6) M elicited negative inotropic one. Either phentolamine or pindolol in a concentration sufficient to block the alpha- and beta-adrenoceptors did not antagonize the positive inotropic effect of ouabain in the papillary muscle preparation of the shrews. PMID- 2568456 TI - Nasal absorption of 17 alpha-ethinyloestradiol in the rat. AB - The bioavailability of 17 alpha-ethinyloestradiol was determined after intravenous, nasal, and intraduodenal, administration in the rat. The results showed that the bioavailability of nasally-administered and intraduodenally administered drug was 80-85, and 15-25%, respectively, of that following intravenous administration. The study demonstrates that the nasal route for the administration of 17 alpha-ethinyloestradiol is superior to the oral route in this species, and may thus provide an alternative for the administration of this drug at lower doses. PMID- 2568457 TI - Adenosine, inosine, hypoxanthine, xanthine and uric acid concentrations in the cerebrospinal fluid of unanaesthetized rats. PMID- 2568458 TI - The effect of lactose particle size on the extrusion properties of microcrystalline cellulose-lactose mixtures. AB - Ram extrusion has been used to assess and compare the flow characteristics and quality of wet powder masses formed from mixtures of water with microcrystalline cellulose and two different particle size samples of lactose. The force displacement profiles can reveal poor flow properties. Steady state flow was achieved for the mixture containing fine lactose which produced an extrudate of uniform moisture content at all extrusion rates except the lowest (5 cm min-1). Increasing the lactose particle size significantly altered the extrusion properties of the formulation with forced flow predominating and high extrusion pressures observed. Formulations showing that type of extrusion are to be avoided as they produce poor quality extrudate which may be unsuitable for spheronization. Flow visualization studies showed that problems associated with such mixtures are caused by failure to maintain a constant angle of convergence during extrusion, which is essential for maintenance of steady state flow. With the mixture made with coarse lactose, the extrudate quality could be improved by extruding at high velocities, information that may be of significance in the development of formulations for large-scale production. Differences between formulations may be expressed quantitatively by plotting the apparent shear stress-shear rate relationship. The curves are modified by the particle size of lactose included in the mixtures. Such curves have implications in predicting the suitability of an extrudate, produced under particular experimental conditions, for spheronization. PMID- 2568459 TI - Physiochemical characterization of methotrexate-bovine serum albumin conjugates. AB - Physiochemical parameters have been determined for a series of methotrexate bovine serum albumin conjugates produced using a water-soluble carbodiimide. Drug coupling produces a heterogeneous product consisting of mixtures of albumin and albumin polymers with varying quantities of covalently bound drug. Due to this heterogeneity, the measurement of absolute physiochemical values was not possible but two factors producing change were evident. The water soluble carbodiimide is responsible for major changes by producing polymers that dominate the properties of the conjugates. The changes induced by the covalently attached drug are less dramatic but still appreciable. The results emphasize that both the attached drug and the method of coupling are significant factors in altering the physiochemical properties of proteinaceous drug carriers. PMID- 2568460 TI - An infrared study of tautomerism in acetohexamide polymorphs. AB - Infrared data determined for known polymorphic forms and some new derivatives of acetohexamide and related compounds support the view that acetohexamide polymorphs exhibit keto-enol tautomerism. They indicate that type A polymorphs exist in the enol form, probably stabilized by intramolecular bonding between an O-H and S = O group to form a six-membered ring. Type B polymorphs exist in the keto form with the urea carbonyl group intermolecularly bonded to a sulphonamide N-H. The new evidence disputes previous interpretations of the data. PMID- 2568462 TI - Relaxant actions of khellin on vascular smooth muscle. AB - To clarify the mechanism of the vasodilatory action of khellin on calcium, we have investigated its relaxant action on base line and on K+ and noradrenaline induced contractile tensions in rat aorta smooth muscle and on spontaneous contractile activity of rat portal vein. Khellin relaxed both of these preparations with a similar potency, which suggests a non-specific inhibition of calcium flux, without any difference related to the specific calcium channels. We have also studied the capacity of khellin to interfere with the loading and release mechanisms of caffeine and noradrenaline-sensitive calcium stores in Ca free medium. Khellin's Ca2+ loading reduction may be related with its capacity to inhibit calcium influx. Khellin applied during Ca2+ release also caused relaxation. We propose that this drug may enhance calcium extrusion or sequestration rather than the calcium release mechanism. These actions on calcium influx and intracellular mobilization can contribute to its vasorelaxant action. PMID- 2568461 TI - Evidence for a postsynaptic action of neostigmine on muscarinic receptors in the anococcygeus muscle of the rat. AB - In the rat anococcygeus muscle neostigmine induces atropine-sensitive, significant leftward displacements of the log concentration-response curves to both noradrenaline and 5-hydroxytryptamine. Responses to high K+ were also potentiated by neostigmine. However, high K+ elicited only small and irregular overflows of tritium from [3H]noradrenaline pre-incubated tissues, in contrast to the large overflow elicited by field stimulation. In addition guanethidine blocked responses to field stimulation but not those to high K+. This is consistent with the dose-related responses to high K+ being elicited postsynaptically. These results indicate that postsynaptic muscarinic receptors are involved in the potentiation by neostigmine of rat anococcygeus muscle responses to field stimulation or exogenous agonists, possibly by an action on receptor operated ion channels. Additional support for a postsynaptic site of action comes from the failure of neostigmine to potentiate tension responses to nerve or field stimulation in the chick expansor secundariorum, a muscle which is devoid of postsynaptic muscarinic receptors. PMID- 2568463 TI - On the mechanism involved in the ability of meptazinol to potentiate the effects of sympathetic nerve stimulation. AB - Mouse isolated vas deferens responded to field stimulation (0.1 Hz) with twitch responses which were abolished by alpha beta-methyleneadenosine 5'-triphosphate (0.5 microM) and were potentiated 2 to 3 fold by meptazinol (5-300 microM). Exogenous adenosine 5'-triphosphate (4-30 microM) also caused a twitch response but was unaffected by meptazinol (30 microM) as was the response to phenylephrine. The effect of meptazinol on the electrically-induced twitch was reproducible, fast in onset, easily reversed by washing and was still seen in the presence of prazosin (1 microM), yohimbine (1 microM), propranolol (0.1 microM), atropine (0.1 microM), physostigmine (1 microM), cocaine (1 microM) or desmethylimipramine (0.3 microM) indicating that the mechanism involved does not depend on adrenoceptors, cholinergic mechanisms or blockade of uptake. Mouse isolated atria responded to stimulation (1, 2 or 5 Hz) of their sympathetic nerves via transmural electrodes with chronotropic responses which were abolished by atenolol (5 and 50 microM) but were unaffected by alpha beta methyleneadenosine 5'-triphosphate (0.5 microM). Meptazinol (100 microM) failed to potentiate the responses. It is suggested that meptazinol potentiates the effects of the non-adrenergic non-cholinergic transmitter thought to be involved in the response of the mouse vas deferens to electrical stimulation. PMID- 2568465 TI - In-vivo (+)-[3H]fenfluramine binding to rat brain: biochemical and autoradiographic studies. AB - The in-vivo binding of (+)-[3H]fenfluramine to rat brain regions is saturable, as shown by the inhibition curves obtained by co-injecting increasing concentrations of unlabelled (+)-fenfluramine: at 2.5 mg kg-1 the inhibition of total bound radioactivity was maximal in all regions. The regional distribution of (+) [3H]fenfluramine specific binding sites (hypothalamus greater than striatum = cortex greater than brainstem greater than hippocampus greater than cerebellum) closely parallels the regional distribution of 5-hydroxytryptamine uptake. Computer-assisted quantitative autoradiography confirms these findings. The IC50 of (+)-fenfluramine for inhibition of its binding in-vivo is below 0.25 mg kg-1, compatible with the presence of high affinity sites. While the physiological role of (+)-[3H]fenfluramine binding sites in the brain, particularly in hypothalamic nuclei, is being investigated, it has been found that in-vivo labelling could also be obtained in the periphery, in lung and renal cortex. The possibility that this peripheral binding is due to the presence of blood platelets cannot be ruled out. PMID- 2568464 TI - Enhancement of phosphatidylinositol turnover and cyclic nucleotide accumulation by chronic anethole trithione treatment in rat submaxillary glands. AB - The effect of chronic treatment with anethole trithione (ANTT) on the phosphatidylinositol (PI) turnover and cyclic (c)AMP and cGMP accumulation in rat submaxillary glands (SMG) has been compared with the effect of chronic treatment with atropine and a cholinesterase inhibitor, diisopropylfluorophosphate (dyflos, DFP). Experiments were performed 24, 48 and 24 h after the last dose of ANTT, atropine and dyflos, respectively. ANTT and atropine enhanced carbachol stimulated [32P] incorporation into phosphatidic acid in the SMG slices, while dyflos showed no effect. Pilocarpine-stimulated in-vivo incorporation of [3H]myoinositol into inositol phosphates was significantly enhanced by ANTT, but not by atropine or by dyflos. Phospholipase C-dependent hydrolysis of phosphatidylinositol 4,5-bisphosphate was significantly enhanced by ANTT and atropine, but not by dyflos. Pilocarpine-stimulated in-vivo accumulation of cAMP and cGMP was enhanced by ANTT and atropine, but dyflos reduced cAMP accumulation without affecting cGMP accumulation. The enhancement of PI turnover and cyclic nucleotide accumulation seems to contribute to the development of supersensitivity of the salivary gland caused by chronic treatment with ANTT and atropine, while reduction of cAMP accumulation may be responsible for the subsensitivity caused by dyflos. PMID- 2568466 TI - The bioavailability of a mixed micellar preparation of vitamin K1, and its procoagulant effect in anticoagulated rabbits. AB - We have investigated the pharmacokinetics and procoagulant activity of a new, mixed-micellar preparation of vitamin K1 (MM-K) in male New Zealand White rabbits. Oral administration of MM-K alone caused a significant (P less than 0.01) increase in the plasma concentrations of vitamin K1 as measured by normal phase high-performance liquid chromatography (HPLC). Maximum plasma concentrations of vitamin K1 (450 ng mL-1, range 133-824 ng mL-1) were recorded at 3.3 h (range 3-5 h), and were significantly (P less than 0.05) greater than those seen after administration of an existing polyethoxylated castor oil preparation (PE-K; Konakion), which were 260 ng mL-1, range 198-390 ng mL-1 (tmax 0.8 h, range 0.4-1.2 h). AUC after MM-K (4.6 micrograms mL-1 h-1, range 2.1-6.3 micrograms ML-1 h-1) was also significantly (P less than 0.05) greater than after PE-K (1.6 micrograms mL-1 h-1, range 1.0-2.1 micrograms ML-1 h-1). However, the bioavailability of vitamin K1 after administration of MM-K was poor (9.4%), and there was considerable intra-individual variability between the concentrations of vitamin K1 recorded in the plasma samples. Both preparations of vitamin K1 stimulated clotting factor synthesis in rabbits anticoagulated with the potent and long-acting coumarin, brodifacoum. Maximum stimulation of clotting factor synthesis by vitamin K1 after MM-K was 87%, range 44-124% (%PCA). The maximum was seen later (tmax 12 h) than after PE-K (PCA 82%, range 47-125%; tmax 5 h). However, there was considerable intra-individual variability in response to both MM-K and PE-K.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568467 TI - Effect of histamine H2-receptor antagonists on acute inflammatory of the rat paw oedema. AB - The effect of three histamine H2-antagonists, cimetidine, ranitidine, and loxtidine, on acute rat paw oedema induced by histamine, carrageenan or complete Freund's adjuvant, have been examined. Administered intraperitoneally, all three antagonists inhibited histamine-induced paw oedema dose-dependently in the range 0.5-15 mumol kg-1. The highest dose of cimetidine produced an inhibition of 92% as against 38% with the same dose of mepyramine. Analysis of the concentration effect curves produced IC50 values of 1.66, 5.12 and 12.30 mumol kg-1 for cimetidine, loxtidine, and ranitidine, respectively, on histamine-induced oedema. In carrageenan-induced inflammation 12.3 mumol kg-1 of each of the three drugs produced significant inhibition, whereas in adjuvant-induced inflammation, (acute phase), cimetidine was very active, loxtidine less so and ranitidine inactive. Thus the relative effectiveness of the antagonists (cimetidine greater than loxtidine greater than ranitidine) appears to differ from their known potency relationship (loxtidine greater than ranitidine greater than cimetidine) on H2 mediated effects. We conclude that H2-receptors are involved in the induction of rat paw oedema, especially those induced by histamine and carrageenan, but that their relative effectiveness appears atypical. PMID- 2568468 TI - Effects of bovine serum albumin and recirculation rate on the uptake of propranolol by rat perfused lung. AB - Lungs isolated from 7-week-old rats were perfused with pH 7.4, oxygenated Krebs Ringer bicarbonate buffer solution containing 2.5 micrograms mL-1 of propranolol and 3 to 5% BSA at the recirculation rate of 4 to 16 mL min-1, at 37 degrees C for 60 min. The extent of propranolol metabolism after 60 min was less than 2.3% of the initial load under any in-vitro perfusion condition. Therefore, the amount disappearing from the perfusion medium was considered as being predominantly that taken up by tissue. Under all experimental conditions, perfusate drug level declined bi-exponentially with time. Apparent in-vitro pulmonary clearance of propranolol was not affected by the increase of BSA level from 3 to 5%. When the perfusate BSA level was fixed at 3%, the lowest recirculation rate (4 mL min-1) yielded the smallest clearance (about 0.15 mL min-1 g-1) but almost constant clearance value (about 0.40 mL min-1 g-1) was obtained at the rate ranging from 8 to 16 mL min-1. The tissue to medium concentration ratio of propranolol, after the perfusion with 3 to 5% BSA at the rate of 8 to 16 mL min-1, was approximately 35, whereas that with 3% BSA at 4 mL min-1 was reduced to about 20. The findings suggest evidence for flow-dependent in-vitro pulmonary clearance of propranolol. PMID- 2568469 TI - Analgesic effects of 3-methoxybenzamide in rats. AB - The i.p. injection of 3-methoxybenzamide (3-MBA) in rats produces a dose-related elevation of the threshold for response to a painful stimulus. Metoclopramide, also a substituted benzamide, has analgesic activity that is attenuated by bromocriptine, a dopamine receptor agonist, and by the narcotic antagonist, naloxone, suggesting involvement of dopamine and opiate receptors in the action of this drug. The involvement of these receptors in the analgesic action of 3-MBA has been examined using L-dopa and naloxone. Neither significantly altered the analgesic action. Although the results are preliminary, the analgesic action of 3 MBA would not seem to occur via opiate or dopamine receptors. PMID- 2568471 TI - The availability of diltiazem: a study on the sorption by intravenous delivery systems and on the stability of the drug. AB - The stability and the sorption by intravenous delivery systems of the calcium antagonist diltiazem dissolved into either 5% dextrose or 0.9% sodium chloride solutions have been investigated, under conditions simulating current clinical practice. Static experiments showed an excellent stability and no sorption after 48 h. Dynamic experiments, at a perfusion rate of 20 mg h-1, showed no sorption of the drug by infusion fluid containers, burettes or administration sets. For end-line filters a temporary decrease of the recovered amount of diltiazem was observed but only with the 0.9% NaCl solution. It is concluded that the stability and the sorption of diltiazem offers no problem with regard to clinical efficacy. PMID- 2568470 TI - Marked increases in plasma catecholamine concentrations precede hypotension and bradycardia caused by 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) in conscious rats. AB - Plasma noradrenaline and adrenaline responses to 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT), a selective putative 5-HT1A receptor agonist, have been studied in conscious, freely moving rats. Intravenously administered 8-OH-DPAT caused dose-related and sustained increases in plasma noradrenaline (2-fold) and adrenaline (11-fold) concentrations. Neither metergoline pretreatment (0.5 mg kg 1 i.v.) nor splanchnicectomy had any effect on the noradrenaline and adrenaline elevation caused by 8-OH-DPAT (250 micrograms kg-1 i.v.). The catecholamine responses peaked early but were still present during nadirs in blood pressure and heart rate. The discrepancy between plasma catecholamine and cardiovascular changes raises further questions about the mechanism of action of 8-OH-DPAT and supports other evidence suggesting a role for vagus stimulation in the cardiovascular effects caused by this drug. PMID- 2568472 TI - Diamorphine stability in aqueous solution for subcutaneous infusion. AB - The influence of temperature and concentration on diamorphine stability during storage over 8 weeks has been investigated. Ampoules containing diamorphine hydrochloride in concentrations from 0.98-250 mg mL-1 have been stored at -20, 4, 21 and 37 degrees C for 8 weeks. Their content of diamorphine, 6 monoacetylmorphine and morphine, on measurement by high performance liquid chromatography after 1, 2, 4, 6 and 8 weeks storage changed to slow degradation of diamorphine at all concentrations at temperatures of 4 degrees C and above. This was accompanied by a corresponding increase in 6-monoacetylmorphine and morphine. There was an associated fall in pH and development of a strong odour characteristic of acetic acid. Precipitation and a white turbidity seen in solutions of 15.6 mg mL-1 and above, appeared after 2 weeks incubation. PMID- 2568473 TI - Evidence for alpha 1-adrenoceptor subtype predominance in the rat seminal vesicle. AB - Noradrenaline (0.6-19 microM) and phenylephrine (2-130 microM) induced contractions in the rat seminal vesicle that were competitively antagonized by the alpha 1-adrenoceptor-selective antagonist corynanthine (120-920 nM). Yohimbine (60-450 nM), an alpha 2-adrenoceptor-selective antagonist, produced a non-competitive antagonism of noradrenaline responses, suggesting that the responses were not alpha 2-adrenoceptor mediated. It is concluded that the rat seminal vesicle has a predominance of alpha 1-adrenoceptors. PMID- 2568474 TI - Contribution of the tissue angiotensin converting enzyme to the antihypertensive effect of altiopril calcium (MC-838) in spontaneously hypertensive rats. AB - The effect of a new orally active angiotensin converting enzyme (ACE) inhibitor, calcium (-)-N-[(S)-3-[(N-cyclohexylcarbonyl-D-alanyl)thio]-2-methylpropionyl+ ++] L- prolinate (MC-838, altiopril calcium), on systemic blood pressure (SBP) and tissue ACE activity has been examined in conscious spontaneously hypertensive rats (SHRs). MC-838 (3 mg kg-1) given orally to SHRs elicited a long-lasting hypotension lasting over 24 h. With the development of the hypotension, MC-838 significantly reduced ACE activity in the lung, kidney and aorta, but not in the brain and heart. Suppression of plasma ACE and rise of plasma renin activity occurred only transiently at an earlier stage. PMID- 2568475 TI - Effects of opioid agonists on urine production in neonatal rats. AB - The modulatory effects of opioids on urine production in adult rats have been well-documented. We report here the first investigation of the effects of these agents on urination in neonatal rats. The kappa-agonists U50,488H (1,10 mg kg-1) and (+)-tifluadom (10 mg kg-1) produced an increase in urine output in 10-day old pups whereas the (-)-isomer of tifluadom was ineffective in this model. The diuretic effects of the highest dose of U50,488H were attenuated by a 10 but not a 1 mg kg-1 dose of the opioid antagonist naltrexone. These findings suggest that kappa-agonists, as in adult animals, produce diuresis in neonates by activity at kappa-opioid receptors and also confirm the stereoselective nature of the response. The increase in urination produced by U50,488H (10 mg kg-1) was also reduced by the alpha-adrenoceptor antagonist phentolamine (1 mg kg -1), an observation which supports the hypothesis that kappa-agonists--in addition to their well-established inhibitory effects on the release of antidiuretic hormone- may increase urination via an adrenergic mechanism at the level of the adrenal medulla. The mu-opioid agonist morphine (0.1-10 mg kg-1), in contrast to its observed effects in older animals, did not produce antidiuresis in either normally-hydrated or water-loaded 10-day old rat pups. The results of this study therefore show that the stimulatory effects of kappa-agonists on urine production appear to be fully-functional at 10-days but the inhibitory effects of opioids on urination lag behind in development. PMID- 2568476 TI - Effects of indeloxazine hydrochloride on cognitive disturbance in cycloheximide treated mice. AB - The effects of indeloxazine hydrochloride [(+/-)-2-[(inden-7 yloxy)methyl]morpholine hydrochloride] on cognitive disturbance in mice subjected to cycloheximide have been examined. Indeloxazine ameliorated cycloheximide induced amnesia in mice, indicating a facilitatory effect of this drug on cerebral functions. Piracetam, Cahopantenate, dihydroergotoxine and viloxazine did not show significant effect on the amnesia. These results suggest that indeloxazine possesses a wider pharmacological profile than piracetam, Cahopantenate, dihydroergotoxine and viloxazine in anti-amnesic activities. PMID- 2568477 TI - Employment of magnet-susceptible microparticles for the targeting of drugs. AB - It has been demonstrated in cats that magnet-susceptible microspheres and liposomes containing neuromuscular blocking agents (dipyronium, pyrocurinum and diadonium) caused a deeper inhibition of the neuromuscular transmission in the limb placed in the magnetic field than in the control limb located beyond the field. The microparticles containing a short-acting neuromuscular blocking agent diadonium appeared to have the highest selectivity of action. The present method allows a pronounced neuromuscular block in a target area to be achieved without noticeable effect on PCO2 of the exhaled air. PMID- 2568478 TI - [Synthesis of 2-oxooctahydroimidazo[1,2-alpha]pyridine-3-spiro-4'-piperidine derivatives possessing antiapomorphine activity]. AB - 4'-Carbamoyl-1,4'-bipiperidine 1 was dehydrogenated on Pd-C to give 2-oxo 2,3,5,6,7,8-hexahydroimidazo[1,2-alpha]pyridine-3-spiro-4'-p iperidine 2, which was reduced to 2-oxo-1,2,3,5,6,7,8,8a-octahydroimidazo[1,2-alpha]pyridine-3-spiro -4'- piperidine 3 with NaBH4. The iminodibenzyl and similar derivatives of 2 and 3 were synthesized and evaluated by using antiapomorphine test in mice. The derivatives of 3 had more potent antagonistic activity than those of 2 and the order of potency of 3 was: chloriminodibenzyl greater than chlordibenzocycloheptene greater than iminodibenzyl greater than iminostilbene greater than fluorene. Further chemical modification of the most active chloriminodibenzyl derivative 8, such as the substitution of Cl atom to Br atom, N-methylation on amine moiety, the replacement of imidazopyridine ring to imidazopyrrole ring, did not give any positive effect. Therefore, 8 may have potential usefulness as an antipsychotic drug. PMID- 2568479 TI - Chronic high velocity projectile injury to the foot. AB - Acute projectile injuries to the foot can present a challenge for the podiatric physician, especially in terms of their chronic effects. The case of a shrapnel wound to the right foot and ankle that resulted in recurrent episodes of soft tissue infection and disability is presented. Treatment consisted of excision of the shrapnel fragment, debridement, and primary closure of the sinus tract created by the projectile. The authors discuss the acute and chronic effects of projectile injuries, factors responsible for determining the severity of these wounds, and various methods of treatment. PMID- 2568480 TI - Tolerance to respiratory actions of sufentanil: functional tolerance and route dependent differential tolerance. AB - The work was aimed at 1) characterizing the development of chronic tolerance to the respiratory effects of sufentanil in anesthetized and awake rats and 2) analyzing the influence that routes of administration of opiates can exert on tolerance development and expression. Tolerance was developed by infusing sufentanil s.c. with osmotic minipumps for 7 days; it was evaluated by injecting acute sufentanil while the minipumps were still maintained. When maximal depression attained was considered, dose-response curves were displaced to the right, the tolerance indexes being 7.3 and 2.6 in awake and anesthetized rats, respectively. No displacement was observed when acute sufentanil was injected i.c.v. and i.v., indicating lack of tolerance. However, several signs of tolerance were detected when the areas under the time-course curve of the effects were considered: faster and more complete recovery of the depression and flattening of the slope of the dose-area under the curve responses, even after i.c.v. and i.v. administration. The results indicate the need for distinguishing between development and expression of tolerance. Differential tolerance can be explained by considering tolerance expression as a deferred phenomenon. In addition, functional compensation stands as a factor of tolerance and as a mechanism responsible for dose-response curve flattening in tolerant subjects. PMID- 2568481 TI - Catecholamine receptors regulating serotonin N-acetyltransferase activity and melatonin content of chicken retina and pineal gland: D2-dopamine receptors in retina and alpha-2 adrenergic receptors in pineal gland. AB - The type of catecholamine receptor involved in the regulation of serotonin N acetyltransferase (NAT) activity and melatonin (N-acetyl-5-methoxytryptamine) level in chicken retina was compared to that regulating these parameters in the pineal gland. Systemic administration of apomorphine, a dopamine receptor agonist, resulted in marked inhibition of the nocturnal increase of retinal NAT activity and melatonin content. Apomorphine did not affect NAT activity or melatonin content of the pineal gland. In contrast, clonidine, an alpha-2 adrenergic receptor agonist, inhibited the nocturnal rise in pineal NAT activity and melatonin content although being without effect on these parameters in retina. Apomorphine-induced inhibition of retinal NAT activity was blocked by spiperone, a D2-dopamine receptor antagonist, but not by antagonists of D1 dopamine, alpha-1, alpha-2 and beta adrenergic receptors. Systemic or intraocular injection of quinpirole, a D2-dopamine receptor agonist, in the middle of the dark phase of the light-dark cycle markedly reduced retinal NAT activity and melatonin level, whereas injections of SKF 38393-A, a D1-dopaminergic agonist, had no effect. The inhibitory effect of clonidine on pineal NAT activity was blocked by yohimbine, an alpha-2 adrenergic receptor antagonist. The results presented in this paper demonstrate that NAT activity and melatonin content in chicken retina and pineal gland are differentially modulated in vivo by D2 dopamine and alpha-2 adrenergic receptors, respectively. Despite the different types of receptors involved, both tissues may share a common pathway for catecholamine-mediated inhibition of melatonin biosynthesis, i.e., inhibition of adenylate cyclase activity. PMID- 2568482 TI - First Oral Medicine World Workshop - June 1988. PMID- 2568483 TI - Loss of Harvey ras heterozygosity in oral squamous carcinoma. AB - This investigation of oral squamous carcinoma in five individuals revealed that four of the patients were constitutionally heterozygous at the c-Ha-ras-1 locus and that the tumour from one patient had lost that heterozygosity. The loss of c Ha-ras-1 alleles provides a useful marker for detecting deletions of genetic material located on the short arm of chromosome 11 (11p) and has been found in association with a number of malignant tumours but has not been previously described in carcinoma of the head and neck. The repeated association of 11p deletions with malignancies has led to the postulation of a recessive cancer gene or tumour suppressor gene at this location involved in carcinogenesis and tumour progression. This study indicates that such a mechanism may contribute to the development of oral squamous carcinoma. PMID- 2568484 TI - Flexural strength of a high-temperature soldered cobalt-chromium alloy. AB - The flexural strength of a high-temperature soldered cobalt-chromium alloy was compared with that of the intact alloy. In one test group the specimens were soldered by Vitallium welding rods and in the other group Bego cobalt-chromium solder was used. The cylindrical portion of all specimens was mounted equally in an Instron testing machine. No statistically significant differences were found in flexural strength between the groups. PMID- 2568485 TI - Autoradiographic visualization of kappa opioid receptors with labelled dynorphins in guinea pig brain. AB - The distribution of kappa opioid receptors in guinea pig brain was measured by in vitro receptor autoradiography using [3H]dynorphin A1-9, [3H]dynorphin A1-8 and [3H]bremazocine as ligands. The sites labelled by the two dynorphins had identical, heterogeneous distributions in brain sections. High levels of kappa receptors were seen in striatum, claustrum, nucleus accumbens and laminae V and VI of the cerebral cortex. The substantia nigra and superior colliculus also had high dynorphin binding levels. The [3H]dynorphin autoradiographs were closely similar to those obtained using [3H]bremazocine in the presence of mu and delta receptor displacers. It is concluded that tritiated dynorphin A fragments can be used for autoradiographic studies of kappa opioid receptors in brain. PMID- 2568486 TI - Familial polyarteritis nodosa: a serologic and immunogenetic analysis. AB - Familial polyarteritis nodosa (PAN) is a rarely described entity. We describe a family with 2 members with PAN after a common hepatitis B infection. Many other autoimmune diseases and autoantibodies were found in other family members not corresponding to HLA phenotypes, suggesting other non-HLA-linked genetic influences may be operative in predisposition to PAN. PMID- 2568487 TI - Concentrations of sulfasalazine and its metabolites. PMID- 2568488 TI - Immunogenetics of rheumatoid arthritis. PMID- 2568489 TI - Polyarteritis nodosa and familial Mediterranean fever: a report of 2 cases and review of the literature. AB - Two cases of polyarteritis nodosa (PAN) in patients with familial Mediterranean fever (FMF) are reported. These and another 11 cases found in the literature suggest that PAN occurs more commonly in patients with FMF than would be expected in the general population. Perirenal hematoma, which is surprisingly high in patients with FMF, is a life threatening complication of PAN. The diagnosis of PAN in patients with FMF may be delayed due to the similarity of the clinical manifestations of both diseases. PMID- 2568491 TI - Regulatory T-cell function in primary humoral immunodeficiency states. AB - The function of B cells and highly purified T-cell subpopulations was studied in patients with various primary humoral immunodeficiencies. In the presence of normal T4+ cells B cells of the patients with X-linked agammaglobulinemia (XLA) were not capable either for IgM or IgG synthesis, whereas B cells of the patient with hyper-IgM syndrome and three of eight patients with common variable immunodeficiency (CVI) could produce normal amounts of IgM, but not IgG. All patients with XLA and the patient with hyper-IgM syndrome exhibited normal helper function by T4+ cells for both IgG and IgM synthesis of normal B cells. The function of CVI T4+ cells was variable. When the function of the patients' T8+ cells was tested against normal B cells, one CVI patient had overactive suppression capacity; in other patients the suppressor cell activity was normal. The findings indicate considerable heterogenicity in the function of lymphocyte subpopulations of the CVI patients. PMID- 2568490 TI - T cell receptor beta chain polymorphisms are associated with cystic fibrosis. AB - The BglII polymorphism near the constant region of the T cell receptor beta chain (TCR c beta) has been investigated in normal controls, patients with cystic fibrosis (CF), and CF carriers. A significant increase was found in the frequency of the 10.0:9.2 kb heterozygous genotype in the CF carrier group (71%) as compared with normal controls (44%) (p = 0.005). Patients with CF also showed an increased frequency of the heterozygous genotype but this was not significant. These results represent a previously unreported disease association and suggest that there may be some form of epistatic interaction between the CF gene and the TCR beta genes such that the double heterozygote is immunologically advantaged. PMID- 2568492 TI - Interaction between the release of adenosine and noradrenaline during sympathetic stimulation: a feed-back mechanism in rat heart. AB - Interactions between the release of adenosine and noradrenaline were studied during sympathetic stimulation in rat heart perfused in situ. Cardiac sympathetic nerves were activated by electrical stimulation of the left cervicothoracic ganglion, and endogenous noradrenaline and adenosine were measured in the effluent from the heart. Following the onset of a continuous stimulation (6 min) a rise of heart rate was observed which was accompanied by the release of noradrenaline and adenosine. Specific blockade of adenosine receptors by 8 phenyltheophylline enhanced the stimulation induced release of noradrenaline suggesting an effective suppression of the noradrenaline release by endogenous adenosine. Heart rate and the release of adenosine were reduced by the beta 1 adrenergic antagonist bisoprolol, while noradrenaline overflow increased. These results are compatible with the concept of a negative feed-back regulation of noradrenaline release by endogenous adenosine from the stimulated cardiomyocytes. In order to characterize the subtype of the presynaptic adenosine receptors involved, the inhibitory potency on stimulus induced noradrenaline release of metabolically stable adenosine agonists was tested. The order of potency (Cyclohexyladenosine greater than or equal to R-phenylisopropyl-adenosine greater than N-ethylcarboxamidoadenosine greater than S-phenylisopropyl-adenosine) suggests an adenosine A1-receptor mediated presynaptic inhibition of noradrenaline release. PMID- 2568493 TI - Neuropeptide Y-like immunoreactivity in relation to the distribution of sympathetic nerve fibers in the heart conduction system. AB - The distribution of nerve fibers showing neuropeptide Y-like immunoreactivity (NPY-LI) was compared with the distribution of the sympathetic nerve fibers in all parts of the conduction system and adjacent atrial and ventricular tissue of the bovine heart. Tyrosine hydroxylase (TH) and dopamine-beta-hydroxylase (DBH) served as markers for the sympathetic nerve fibers. NPY-LI was detected in most of the sympathetic nerve fibers that were present in nerve fascicles and that were associated with conduction cells and arterial walls in all regions examined. This phenomenon was more apparent when staining for NPY was compared with staining for DBH than with staining for TH. It was also found that some nerve varicosities exhibiting NPY-LI in association with arterial walls and local ganglia did not show DBH-LI. Furthermore, some of the ganglionic cells located in the regions of the conduction system showed NPY-LI but not DBH- or TH-LI. The observations are discussed in relation to what is known of the subcellular localization of NPY, TH and DBH. It is furthermore concluded that an NPY-like peptide is present in the sympathetic innervation of all parts of the conduction system and ordinary myocardial tissue, but that this peptide is also present in nerve fibers in the heart that do not represent sympathetic fibers. The observations raise important questions for further research aimed at determining the effects of NPY in relation to the function of the conduction system and in relation to the functions of not only sympathetic but also non-sympathetic nerve fibers in the heart. PMID- 2568494 TI - Investigators seek to increase taxol supply. PMID- 2568495 TI - Isolation and structural characterization of the bovine tyrosine hydroxylase gene. AB - A bovine tyrosine hydroxylase (TH) cDNA probe was used to screen a charon 30 genomic library. Screening of approximately 1 million recombinant phage resulted in the identification of one clone, lambda B1, containing the entire bovine TH gene. Results derived from restriction endonuclease mapping and sequence analysis reveal that the bovine gene contains 13 exons spanning approximately 7 kb of genomic DNA. Determination of the transcription initiation site indicates that the TH gene has a 5' untranslated region of 27 bp. A TATA-box sequence is located between positions-29 and -24 from the transcription initiation site and a cyclic AMP regulatory element (CRE) between-45 and -38. Although the TH gene appears to be glucocorticoid responsive in vitro, no regions bearing identity to the consensus sequence for the glucocorticoid regulatory element (GRE) were detected within approximately 1.5 kb of 5' flanking sequence. A cross-species comparison of the 5' flanking sequences of the bovine, rat, and human TH genes reveals strong sequence and positional conservation of seven sequence elements. An analysis of the nucleotide sequence within these elements reveals similarity to the consensus sequences reported for known cis-acting regulatory elements and transcription factor binding sites, suggesting that they may play a role in the regulation of TH gene expression. PMID- 2568496 TI - HLA-DR beta, -DQ alpha, and -DQ beta restriction fragment length polymorphisms in multiple sclerosis. AB - Restriction fragment length polymorphism (RFLP) studies were performed on DNA from unrelated Caucasian patients with multiple sclerosis (MS) using cDNA probes to the HLA class II genes DR beta, DQ alpha, and DQ beta. In a study of 34 patients and 34 controls who were not matched for DR type, we found that the DQ beta allele-specific RFLP or allogenotype, termed DQ beta lb, which corresponds at the molecular level to the DQwl serotype, is preferentially associated with MS. A significant disease association with DR2 was demonstrated by serology but this was not confirmed using DR2/Dw2-specific RFLPs. We suggest that DQ beta lb is largely responsible for HLA-associated susceptibility to MS and that the apparent MS-DR2 serological association is due to the strong linkage disequilibrium between DR2 and DQ beta lb. Homozygosity of one of the two allelic bands of the DX alpha gene, usually termed the DX alpha lower (DX alpha L) band (which cross-hybridizes with the DQ alpha probe), correlated with reduced susceptibility to MS. Similarly a 5.3 kb band identified by the DQ alpha probe in Mspl digests showed a negative correlation with MS. In an analysis of 27 DR2+ controls and 26 DR2+ patients it was found that these individuals all had DR2/Dw2 specific RFLPs and all had identical DR2/Dw2-associated DQ beta (DQ beta lb) and DQ alpha (DQ alpha lb) allogenotypes. We detected no polymorphisms of DR beta, DQ alpha, or DQ beta genes among the DR2+ MS patients which distinguished them from normals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568497 TI - Vasoactive intestinal peptide (VIP), a putative neurotransmitter of nonadrenergic, noncholinergic (NANC) inhibitory innervation and its relevance to therapy. AB - Nonadrenergic, noncholinergic (NANC) innervation, the third division of the autonomic nervous system, has both inhibitory and excitatory parts. The excitatory part received only limited attention. Substance P has been suggested to be the neurotransmitter of the excitatory part. The NANC-inhibitory innervation has recently been studied in detail. Although the neurotransmitter has not been conclusively identified, a substantial body of evidence exists to support vasoactive intestinal peptide (VIP) as the neurotransmitter. VIP is widely distributed in the body. Reports show that this innervation in animals and man plays a significant role in both health and disease. Pathological conditions could result from either an increase or decrease in VIP production. An absence of VIP-producing neurons has been identified to be responsible for Hirschsprung's disease in the alimentary system and hyperactive airways in the respiratory system. An increase in VIP production is associated with chronic water diarrhea syndrome in humans. Taking these factors into consideration, various therapeutic measures are suggested with the use of VIP or its antagonists. PMID- 2568498 TI - Sedative and analgesic effects of medetomidine in dogs. AB - The sedative and analgesic effects of medetomidine were studied in 18 laboratory beagles in a randomized cross-over study which was carried out in a double-blind fashion. Xylazine was included as a positive control and placebo as a negative control. Medetomidine was used at doses of 10, 30, 90 and 180 micrograms/kg i.m. compared to a dose of 2.2 mg/kg xylazine i.m. Parameters closely related to sedation were used to measure the degree of sedation. These were a posture variable (including evaluation of the dog's posture without external disturbance and resistance when laid recumbent) and a relaxation variable (including relaxation of the jaws, upper eyelids and anal sphincter). The first signs of sedation were recorded 1.5-3.5 min after administration of both drugs. The dogs sat down at 0.6-2.6 min post-injection and became prone at 1.9-5.9 min. Medetomidine dose-dependently affected the posture of the dogs and the relaxation variable--the higher the dose, the stronger and longer lasting the effect recorded. The sedative effect of xylazine was comparable to a medetomidine dose of 30 micrograms/kg. The analgesic effect was assessed as changes in the response to superficial pain induced by electrical stimuli. The response threshold increased significantly with both drugs and the effect of medetomidine was dose dependent. The effects of the doses of 30 micrograms/kg medetomidine and 2.2 mg/kg xylazine did not differ significantly. In summary, medetomidine possessed an excellent sedative effect associated with analgesia in dogs. PMID- 2568499 TI - Pharmacological responses of rabbit urinary bladder after subtotal cystectomy. AB - Partial cystectomy is used clinically in specific circumstances. There have been some reports indicating that the bladder following subtotal cystectomy can regenerate to a certain degree. The present study investigates the physiology and pharmacology of bladder regeneration at eight weeks after resection of a major part of the bladder body in rabbits. The following studies were performed on control and cystectomy bladders: 1) in vivo cystometry (prior to and four weeks after the operation, and before the sacrifice at eight weeks); 2) sequential muscle strip study (the strips were obtained by dividing bladder transversely into upper body, lower body, mid-bladder, upper base and lower base); and 3) in vitro whole bladder studies. The results can be summarized as follows: 1) grossly there was no regeneration of the bladder body; the cystometric capacity was increased slowly after the operation primarily due to increased size and capacity of the bladder base. 2) The contractile response of the subtotal cystectomized bladder body to field stimulation and bethanechol stimulation was less than the response of normal bladder body. 3) The contractile response to epinephrine showed that the ratio of alpha/beta adrenergic response was much greater in the cystectomy bladder than in the normal bladder. 4) In the whole bladder study, the intravesical pressure response to field stimulation was about the same in both groups, the response to bethanechol was less for cystectomized bladder, and the response to methoxamine was greater for the cystectomized bladder. 5) The ability of cystectomy bladder to empty in response to both field stimulation and bethanechol was impaired whereas the control bladder fully emptied to both field stimulation and bethanechol. In conclusion, over the eight week period following subtotal cystectomy the capacity increased via distention (and hypertrophy) of the bladder base area as opposed to regeneration of the bladder body. PMID- 2568500 TI - Histologic analysis of orchiopexy in a cryptorchid rabbit model. AB - Although there is a consensus that very early orchiopexy is advantageous, there is little experimental data to support this view. Therefore, we studied early pre pubertal versus later pre-pubertal orchiopexy in a cryptorchid rabbit model. Male offspring were divided into four groups with six to nine rabbits in each group: 1) controls (CON); 2) shams undergoing sham operations at both three weeks and two months of life (SHAM); 3) cryptorchid on day 3 with orchiopexy at three weeks and sham operation at two months (O3W); 4) cryptorchid on day 3 with sham operation at three weeks and orchiopexy at two months (02M). All animals were sacrificed post-puberty at six months of age; the tests were fixed in situ by perfusion fixation, embedded in glycol methacrylate, and underwent morphometric analysis in a blinded fashion. The average seminiferous tubular diameter (STD) of the CON, SHAM, O3W, and O2M groups was 234, 160, 116, and 123 microns, respectively. The average testicular biopsy score count (TBSC) of the CON, SHAM, O3W and O2M groups was 9.0, 5.2, 3.5 and 3.5, respectively. There is no significant difference in STD or TBSC between the O3W and O2M cryptorchid groups, and values for both of these groups were significantly different from those of the SHAM group. In addition, a significant effect of sham operation was demonstrated. These results demonstrate no advantage of very early orchiopexy in this model, and are consistent with the clinical literature which, in general, shows reduced reproductive potential of a cryptorchid testis after orchiopexy. PMID- 2568501 TI - Acoustic and plasma guided lasertripsy (APGL) of urinary calculi. AB - The feasibility of using pulsed laser generated acoustic and plasma optical emission signals to monitor laser fragmentation of urinary stones was studied in vitro. A flashlamp pumped tunable dye laser operating at a wavelength of 504 nm. (coumarin green) was used as the laser source. Acoustic signals were recorded with a hydrophone which has a useful frequency response of up to 350 KHz. Plasma optical emissions were transmitted retrograde along the laser fiber and reflected through a beam splitter to an optical detection system consisting of a series of spectral filters (to transmit plasma radiation from 380 nm.-440 nm. and block any 504 nm. laser light) and a photomultiplier tube. Measurements of acoustic and plasma signals were taken from different urinary calculi, guidewires, catheters, blood, blood clots, bruised soft tissue and normal ureter. Signals were also obtained from stones placed in ureter of an ex vivo bovine urinary tract specimen. Results of monitoring both plasma and acoustic signals show that it is possible to determine, without direct vision, whether the laser is hitting stone, ureteral wall or lumen. Strong plasma and strong acoustic signals are produced by calculi; strong acoustic but no plasma signals suggest that the laser is hitting blood clot or bruised ureteral wall. Absent plasma and acoustic response indicate that the laser is discharging on normal ureter or in the lumen. These distinctions may allow clinical stone fragmentation without direct ureteroscopic vision. PMID- 2568502 TI - Postnatal testicular maldevelopment in unilateral cryptorchidism. AB - Histomorphometric analysis of semi-thin sections was performed on testicular biopsies of 232 unilaterally cryptorchid testes and 195 of their contralateral descended partners. The results demonstrated a decreased number of germ cells detectable from the first year of life. There was delayed and defective transformation of gonocytes to Ad spermatogonia, which normally is complete at age 6 months, delayed or failed transformation of Ad spermatogonia to primary spermatocytes, which normally commences at age 3 years and decreased numbers of Leydig cells. These abnormalities were present in the unilaterally cryptorchid testes and their contralateral descended partners but they were more severe, of earlier onset and more progressive in the cryptorchid testes. These findings are compatible with the hypothesis that hypogonadotropic hypogonadism is the cause of the increased incidence of infertility seen in unilateral cryptorchidism. PMID- 2568503 TI - Recovery of testicular blood flow following ligation of testicular vessels. AB - To determine whether initial ligation of the testicular vessels of the high undescended testis followed by a delayed secondary orchiopexy is a viable alternative to the classical Fowler-Stephens procedure, a series of preliminary experiments were conducted in the rat in which testicular blood flow was measured by the 133xenon washout technique before, and 1 hour and 30 days after ligation of the vessels. In addition, testicular histology, and testis and sex-accessory tissue weights were measured in 6 control, 6 sham operated and 6 testicular vessel ligated rats 54 days after vessel ligation. The data demonstrate that ligation and division of the testicular blood vessels produce an 80 per cent decrease in testicular blood flow 1 hour after ligation of the vessels. However, 30 days later testis blood flow returns to the control and pre-treatment value. There were no significant changes in testis or sex-accessory tissue weights 54 days after vessel ligation. Histologically, 4 of the surgically operated testes demonstrated necrosis of less than 25 per cent of the seminiferous tubules while 1 testis demonstrated more than 75 per cent necrosis. The rest of the tubules in all 6 testes demonstrated normal spermatogenesis. From this study we conclude that initial testicular vessel ligation produces an immediate decrease in testicular blood flow but with time the collateral vessels are able to compensate and return the testis blood flow to its normal pre-treatment value. These preliminary observations lend support for the concept that initial ligation of the testicular vessels followed by a delayed secondary orchiopexy in patients with a high undescended testis may be a possible alternative to the classical Fowler-Stephens approach. PMID- 2568504 TI - Orchiopexy: an experimental study of the effect of surgical technique on testicular histology. AB - Orchiopexy was performed in 35 mature Sprague-Dawley rats using absorbable suture, nonabsorbable suture or a classical dartos pouch technique without suture fixation. Examination of the chromic-fixed testes revealed severe acute inflammation of the suture site and most of the parenchyma in 76 per cent, with abscess formation in 65 per cent, complete absence of spermatogenesis in 82 per cent and evidence of tubular necrosis in 88 per cent. Nylon-fixed testes were adherent only at the sutures, with absent spermatogenesis in 29 per cent, moderate to severe tubular necrosis in 29 per cent and tubular atrophy in 58 per cent. Dartos-fixed testes demonstrated complete circumferential adherence, normal spermatogenesis in 94 per cent and minimal focal tubular atrophy in 23 per cent. Contralateral testes appeared normal in all animals. PMID- 2568505 TI - Significance of epididymal and ductal anomalies associated with testicular maldescent. AB - Epididymal abnormalities have long been found in conjunction with cryptorchid testicles. The aim of this study was to document the different types of epididymal and ductal anomalies in relation to the position of the cryptorchid testis and to determine their clinical significance. Exploration was performed on 187 cryptorchid testes in 174 boys. The position of the testis and the epididymal and ductal anomalies were documented at operation. A biopsy was taken from the testis or atretic spermatic cord structures for light microscopy. Epididymal, ductal and/or testicular anomalies were detected in 43 per cent of the 187 testes, including anomalies of ductal fusion, anomalies of ductal suspension and anomalies associated with absent or vanishing testes. Biopsy of the testes with severe anomalies of ductal fusion showed preservation of germ cells in 69 per cent and diminished germ cells in 31 per cent. The higher the arrest of testicular descent, the more grossly abnormal was the associated ductal system. Early successful orchiopexy alone may not ensure subsequent fertility despite the presence of normal germ cells. PMID- 2568506 TI - Cryptorchidism, orchiopexy and infertility: a critical long-term retrospective analysis. AB - We assessed the fertility of 40 patients who underwent orchiopexy between 1950 and 1960. Testicular biopsies also had been performed at operation, a practice exceptional for the era. Of 23 patients with unilateral undescended testes who attempted to have children 20 (87 per cent) were successful. In contrast, only 3 (33 per cent) of 9 patients with bilateral undescended testes fathered children. Over-all sperm counts in 16 patients were low but they were not predictive of paternity. Testicular biopsy specimens were reviewed and the fertility index was determined. In most cases a good correlation was found between histological status and paternity status. PMID- 2568507 TI - Peace through mind/brain science. PMID- 2568508 TI - A single nocte dose of famotidine in the treatment of duodenal ulcer. AB - Fifty three patients with duodenal ulcer were treated with 40 mg of famotidine at bed time. Repeat endoscopy showed healing rates of 69.8% and 83% at 4 and 6 weeks respectively. Seven patients were kept on a maintenance dose of 20 mg nocte for one year, of these one had a relapse despite taking the drug regularly. No appreciable side effects were observed with the drug (JPMA 39:104, 1989). PMID- 2568509 TI - [Molecular genetics and DNA diagnosis for inherited coagulation disorders]. PMID- 2568510 TI - [Clinical investigation of urinary activity of NAG and AAP in patients with various types of primary glomerulonephritis]. AB - We investigated the activity of NAG and AAP in the urine of 36 patients with chronic glomerulonephritis (GN) with special reference to the relation between the levels of the enzymes and histopathological changes of the kidney. The subjects consist of 7 cases of minimal change nephrotic syndrome (MCNS), 3 cases of membranous GN, 5 cases of IgA. GN, 5 cases of membranoproliferative GN, 3 cases of focal glomerulosclerosis and 12 cases of chronic renal failure who had creatinine levels of more than 2 mg/dl resulting from chronic GN. Urinary levels of NAG and AAP were significantly higher in cases with GN than those in cases with chronic renal failure. In cases with GN, urinary levels of NAG and AAP correlated with the amounts of urinary protein, while those levels had no correlation with the classification of histopathological changes of glomeruli. The high value of NAG/mg.protein ratio, NAG by urinary protein, might be explained by the degree of tublointerstitial damage. We concluded that urinary levels of NAG and AAP reflects the proximal tubular damage caused by protein reabsorption. PMID- 2568511 TI - Pharmacologic characterization of a novel non-benzodiazepine selective anxiolytic, DN-2327. AB - DN-2327, 2-(7-chloro-1,8-naphthyridin-2-yl)-3-[(1,4-dioxa-8- azaspiro[4.5]dec-8 yl)carbonylmethyl]isoindolin-1-one, produced anxiolytic, taming and anti convulsive effects when administered orally to several species of animals. DN 2327 produced few of the sedative-hypnotic and muscle-relaxant effects observed with diazepam. The durations of the anxiolytic and anti-convulsive activities of DN-2327 were much longer than those of diazepam. Tolerance to DN-2327 did not develop when it was administered daily for 14 days in an anti-conflict test (Vogel conflict test). DN-2327 showed potent displacement activity against [3H]diazepam binding. The binding affinity of DN-2327 for benzodiazepine receptors was about twenty times that of diazepam. Furthermore, the affinity of DN-2327 for benzodiazepine receptors was not enhanced by the presence of GABA. There is a wide margin between the doses of DN-2327 that cause the anxiolytic effects and its sedative-hypnotic/muscle-relaxant effects. These results suggest that DN-2327 has more marked anxioselective properties compared with the benzodiazepines. PMID- 2568512 TI - Course of cerebral lesions in a patient with periarteritis nodosa studied by magnetic resonance imaging. AB - The course is reported of a patient with periarteritis nodosa who initially presented with neurological symptoms. Multiple cerebral lesions were documented by the first magnetic resonance imaging (MRI) investigation. The majority of these had disappeared completely in the follow-up MRI studies. In contrast to neurological improvement the patient eventually died due to multiorgan failure. Postmortem histological examination revealed no pathological findings in the brain except one single necrotic area already known from MRI. Remissions of histological and angiographic alterations in periarteritis nodosa have been described as "local healing" leading to fibrosis and scarring. Our findings suggest that restitutio ad integrum may occur, at least in cerebral lesions. PMID- 2568513 TI - [A case of periarteritis nodosa]. PMID- 2568514 TI - [Do not be afraid to administer analgesics! Patients with cancer feel pain]. PMID- 2568515 TI - Changes in neuronal size and neurotransmitter marker in hereditary canine spinal muscular atrophy. AB - Hereditary canine spinal muscular atrophy (HCSMA) is a dominantly inherited motor neuron disease that produces muscle weakness and atrophy. Immunocytochemical and computer-imaging morphometric methods were used to compare early changes that occurred in dogs with HCSMA (n = 4) versus controls (n = 2). The size and number of neurons in the ventral horn and the number of motor neurons expressing choline acetyltransferase were quantitated. The density of all ventral horn neurons per micrometer squared in dogs with HCSMA was greater than controls, and there were more small neurons than in controls. Immunocytochemical methods revealed more small cholinergic neurons and fewer large cholinergic neurons in HCSMA than in controls, suggesting growth arrest in HCSMA or a shift in size class from large cholinergic neurons to small ones. The density of cholinergic neurons per micrometer squared was not significantly different between the two groups. Analysis of predicted distributions of cholinergic and noncholinergic neurons revealed that HCSMA cholinergic neurons were smaller and that, in some size classes, fewer neurons expressed choline acetyltransferase. These observations indicate that in HCSMA the motor neuron fails to achieve normal size and/or undergoes atrophy. PMID- 2568517 TI - [Usefulness of biological markers in the detection of alcoholism in primary care situations]. PMID- 2568516 TI - Epirubicin cytotoxicity but not oxygen radical formation is enhanced by four different antiemetics. AB - The anthracycline epirubicin (0.1-1.0 mg l.-1) caused a dose-dependent inhibition of the clonogenic survival of Chinese hamster fibroblasts. Four antiemetics (dixyrazin, metoclopramide, chlorpromazine and droperidol) augmented the inhibition by epirubicin. Furthermore, by using oxygen consumption as an index, epirubicin-induced free oxygen radical formation was not potentiated by the tested antiemetics. The results justify further studies on the antineoplastic and adverse effects pertaining to interaction between anthracyclines and antiemetics. Finally, the results suggest that interference of antiemetics on fibroblast toxicity does not directly involve the generation of free oxygen radicals. PMID- 2568518 TI - Nipradilol, a beta-adrenoceptor antagonist having a vasodilatory action, attenuates myocardial acidosis induced by coronary artery occlusion in dogs. AB - In dogs anesthetized with pentobarbital, the anterior descending coronary artery (LAD) was partially occluded to reduce LAD flow to about half of the original flow (partial occlusion). Myocardial pH (MpH) was measured by the use of a micro glass pH electrode. MpH decreased from 7.5-7.63 to 6.82-6.86 30 min after partial occlusion of the LAD. Nipradilol (0.3 mg/kg) was injected intravenously 30 min after partial occlusion, which continued for a further 60 min after nipradilol injection. Nipradilol decreased blood pressure and heart rate, and significantly increased myocardial pH which had been decreased by partial occlusion, within 60 min after injection. Nipradilol-induced restoration of the myocardial [H+] (calculated from the pH data), that had been increased by partial occlusion, was 48.5%. Bradycardia induced by nipradilol was not a determinant factor in the pH effect of nipradilol, because even in the paced heart, nipradilol restored the myocardial [H+] that had been increased by partial occlusion. These results indicate that nipradilol attenuates ischemia-induced myocardial acidosis, suggesting the favorable effect of nipradilol on ischemic myocardium. The favorable effect of nipradilol may be due to the beta-adrenoceptor antagonistic effect rather than the vasodilating effect. PMID- 2568519 TI - Randomised trial of fish oil for prevention of restenosis after coronary angioplasty. AB - To examine the potential role of fish oil supplementation in the prevention of restenosis after coronary angioplasty (PTCA), a randomised double-blind trial was conducted in 204 patients. The treatment group received 6 g/day of n-3 fatty acids, beginning 5.4 (SD 3.2) days before PTCA, and continuing for 6 months; the control group received olive oil placebo. Compliance was assessed by pill count and plasma levels of eicosapentaenoic acid (EPA). Restenosis was identified by symptoms and exercise testing and confirmed by angiography. PTCA was successful in 186 patients (93%). The incidence of angiographic restenosis was 34% in the fish oil group and 23% in the control group (relative risk 1.7, 95% CI 0.9-3.4). The lack of benefit of fish oil was not influenced by length of pretreatment, compliance with study medication, or the concentrations of plasma EPA achieved. PMID- 2568520 TI - Randomised controlled trial of subcutaneous calcium-heparin in acute myocardial infarction. The SCATI (Studio sulla Calciparina nell'Angina e nella Trombosi Ventricolare nell'Infarto) Group. AB - In a multicentre study 711 patients were randomised to a group receiving calcium heparin, 12,500 U, subcutaneously (360), or to a group receiving no heparin (351), beginning within 24 h of the onset of symptoms. 433 of these patients, admitted within 6 h, were given intravenous streptokinase (SK). Results were analysed for the in-hospital period. Calcium-heparin had no significant effects on the frequency of electrocardiographically documented ischaemic episodes or non fatal reinfarction in the whole series; in the subgroup receiving SK, transient ischaemic episodes recurred in 14.2% of those treated with heparin vs 19.6% of the controls (p = 0.08). Mortality was significantly lower in the calcium-heparin treated patients, both in the overall groups (21/360 vs 35/351, p = 0.03) and in the SK subgroups (10/218 vs 19/215, p = 0.05). In 200 patients with first anterior myocardial infarction, it was possible to assess the effect of heparin on left ventricular mural thrombosis. On predischarge two-dimensional echocardiography, the prevalence of thrombus was significantly lower in the heparin group than in the control group (19/107 vs 34/93). Heparin also greatly reduced the incidence of thrombus formation in those who were thrombus-free on admission. Complications of heparin treatment were few. PMID- 2568521 TI - Effect of a Campylobacter pylori protein on acid secretion by parietal cells. AB - The uptake of [14C]-aminopyrine by rabbit gastric epithelial cells was used as an indirect assay for acid secretion from parietal cells. Campylobacter pylori strains, isolated from the stomachs of 3 patients with chronic gastritis, duodenal ulcer, and near-normal mucosa, respectively, inhibited acid secretion as effectively as 10(-4) mol/l cimetidine. The inhibition occurred with whole organisms and with sonicates. Preliminary characterisation suggested that the inhibition was due to a nondialysable protein. The inhibitor was not toxic to gastric epithelial cells. This bacterial product may explain the hypochlorhydria seen in man acutely infected with C pylori. PMID- 2568522 TI - Endotoxaemia, pulmonary complications, and thrombocytopenia in liver transplantation. AB - Plasma endotoxin was measured in 64 patients undergoing primary liver replacement. Endotoxin concentrations increased during the anhepatic phase of the operations, and remained high for several days. Although the severity of endotoxaemia did not correlate with duration of the anhepatic phase, there was a correlation between endotoxaemia and the need for perioperative platelet transfusions, ventilator dependency postoperatively, and one-month case-fatality. PMID- 2568523 TI - Cultured composite skin grafts: biological skin equivalents permitting massive expansion. AB - After assessment in mice, a biological skin equivalent consisting of cultured fibroblasts in a collagen gel overlain with cultured keratinocytes was applied to three patients who had had tattoos excised. In all patients the grafts took well with good cosmetic results and little or no contracture. A biopsy at 4 weeks showed that the central graft area had a fully differentiated epidermis and a mature dermis without adnexae. PMID- 2568524 TI - Databases for health care outcomes. PMID- 2568525 TI - Oxcarbazepine. PMID- 2568526 TI - Diagnosis of Lyme disease. PMID- 2568528 TI - Economics for medical students. PMID- 2568527 TI - Management of asthma in the community. PMID- 2568529 TI - APACHE-II score for assessment and monitoring of acute pancreatitis. AB - The value of the Acute Physiology and Chronic Health Enquiry (APACHE-II) score, the Simplified Acute Physiology score, and the Medical Research Council (MRC) sepsis score were compared with clinical assessment and Ranson and Imrie scores in the evaluation and monitoring of acute pancreatitis in 290 attacks. Attacks were graded mild (231) if uncomplicated, or severe (59) when major organ failure or a pancreatic collection occurred. Only APACHE-II scores were available at the time of admission; they correctly predicted outcome in 77% of attacks and identified 63% of severe attacks, compared with 44% achieved by clinical assessment. After 48 h, APACHE-II was most accurate, and correctly predicted outcome in 88% of attacks, compared with 69% for Ranson and 84% for Imrie scores. APACHE-II predicted 73% of pancreatic collections at 48 h, compared with 65% for Ranson and 58% for Imrie scores. In acute pancreatitis, APACHE-II may facilitate rapid selection of patients for intensive therapy or clinical trials, improve comparison between groups of patients, and indicate that a pancreatic collection is probable. PMID- 2568530 TI - Sputum induction for diagnosis of Pneumocystis carinii pneumonia. AB - 51 patients infected with the human immunodeficiency virus (HIV) and with suspected Pneumocystis carinii pneumonia underwent sputum induction. Sputum specimens from 18 patients were positive for P carinii and patients were treated appropriately 25 of the 33 patients with negative specimens underwent bronchoscopy and bronchoalveolar lavage within 4 days of sputum induction, only 1 of whom had evidence of Pneumocystis infection. For diagnosis of Pneumocystis carinii pneumonia, sputum induction according to a strict protocol had a sensitivity of 94.7% and a negative predictive value of 96%, compared with bronchoalveolar lavage. PMID- 2568532 TI - Cervical smears: a questionable practice? AB - Screening for carcinoma of the cervix by cervical smears satisfies none of the criteria which would provide its justification. It is an expensive contribution to ill health because the harms exceed the possible benefits by a substantial margin. PMID- 2568533 TI - Campylobacter pylori and acid secretion. PMID- 2568531 TI - Outbreak of beri-beri in The Gambia. AB - In 1988, an outbreak of beri-beri occurred in The Gambia during the rainy season. At least 140 people were affected, and 22 died. A clear response to thiamine was observed in 8 patients who were in hospital, and the outbreak resolved after distribution of thiamine in the community. Gradual replacement of traditional cereals with imported milled rice may have increased susceptibility to beri-beri. PMID- 2568534 TI - Endoscopic drainage for suppurative cholangitis. PMID- 2568535 TI - Pruritus and antimalarial drugs in Africans. PMID- 2568536 TI - Haem and porphyria attacks. PMID- 2568537 TI - Phenotype and function of Japanese adult T-cell leukaemia cells. PMID- 2568539 TI - Sex ratio in singleton and multiple births after infertility treatment. PMID- 2568538 TI - Salt and blood pressure. PMID- 2568540 TI - Two-embryo transfer. PMID- 2568541 TI - "Boil the water". PMID- 2568542 TI - Identification of unsuspected cases of giardiasis by wet-film microscopy. PMID- 2568543 TI - Red blood cell morphology in chronic fatigue syndrome. PMID- 2568544 TI - Treatment of meningioma with octreotide. PMID- 2568545 TI - Diagnosis of enteropathogenic Escherichia coli. PMID- 2568546 TI - Handwashing and horizontal spread of viruses. PMID- 2568547 TI - Detecting t(14;18) in paraffin-embedded lymphoma tissue. PMID- 2568548 TI - Degeneration of junctional tachycardia to pre-excited atrial fibrillation after intravenous verapamil. PMID- 2568550 TI - Boycotting journals. PMID- 2568551 TI - Future of French-language journals. PMID- 2568549 TI - Allegations of torture in Turkey. PMID- 2568552 TI - WHO and PLO. PMID- 2568553 TI - Adverse effect of tamoxifen with LHRH agonist on oestrogen-receptor-negative mammary carcinoma. PMID- 2568554 TI - Insulin autoantibodies as markers of potential diabetes mellitus. PMID- 2568555 TI - Mobilisation after Achilles tendon rupture. PMID- 2568556 TI - Papillomavirus infection and skin cancer in renal allograft recipients. PMID- 2568558 TI - HIV-1 infection in males and females in Central Africa. PMID- 2568557 TI - CSF placental alkaline phosphatase as marker in cranial dysgerminoma. PMID- 2568559 TI - HIV infection, hepatitis, and syphilis in Spanish prisons. PMID- 2568560 TI - Improvement in AIDS patients on peptide T. PMID- 2568562 TI - Lewis negative genotype and breast cancer risk. PMID- 2568561 TI - Rapamycin for immunosuppression in organ allografting. PMID- 2568563 TI - Wilson's disease patients can be decoppered. PMID- 2568564 TI - Biotinidase deficiency associated with severe combined immunodeficiency. PMID- 2568565 TI - Presenile dementia associated with mosaic trisomy 21 in a patient with a Down syndrome child. PMID- 2568567 TI - False-positive HIV western blots. PMID- 2568566 TI - Rapid high dose streptokinase in severe pulmonary embolism. PMID- 2568568 TI - [Clinical and epidemiologic characteristics of hemorrhagic fever with renal syndrome]. AB - Clinical and epidemiological data on 16 patients with HFRS admitted to the University Hospital of Infectious Diseases in Zagreb during the past 10 years (1977-1986) are reported. In 13 of them, the diagnosis was confirmed serologically by indirect fluorescent method. All but two were men between 20 and 45 years of age (80% of patients). The disease appeared sporadically only. According to the domicile, way of living and working all the patients but three mentioned the contact with rodents in the fields or in other places (mill, storehouse). Three patients live in Zagreb and there was no possibility to get infected out of the town. The disease had moderate course, more rarely severe course when shock and acute renal failure developed. General symptoms with fever and algias dominated (febrile stage). Some of the patients had ophthalmic disturbances. Patients with a pronounced gastroenteritis developed shock the most frequently. In some of the patients back and abdominal pains followed by obstipation appeared. HFRS was incriminated disease because of febrile stage followed by the acute renal failure, oliguria, azotemia, polyuria and low urine osmolity. One female patient with active rheumatoid arthritis died after two-week staying in hospital due to irreversible shock followed by cardiopulmonary and renal failure. PMID- 2568569 TI - Stress and immunity: an integrated view of relationships between the brain and the immune system. AB - The old notion that stress exacerbates the progression of physical illness via its corticosteroid-mediated immunosuppressive effects must be revised. Experimental and clinical studies demonstrate that both laboratory and natural stressors alter the activities of lymphocytes and macrophages in a complex way that depends on the type of immune response, the physical and psychological characteristics of the stressor and the timing of stress relative to the induction and expression of the immune event. The influences of stress on immunity are mediated not only by glucocorticoids but also by catecholamines, endogenous opioids and pituitary hormones such as growth hormone. Sensitivity of the immune system to stress is not simply fortuitous but is an indirect consequence of the regulatory reciprocal influences that exist between the immune system and the central nervous system. The immune system receives signals from the brain and the neuroendocrine system via the autonomic nervous system and hormones and sends information to the brain via cytokines. These connections appear to be part of a long-loop regulatory feedback system that plays an important role in the coordination of behavioral and physiological responses to infection and inflammation. PMID- 2568570 TI - The influence of carotid sinus pressure on plasma atrial natriuretic peptide in anaesthetized rabbits. AB - The effect of changes in carotid sinus perfusion pressure on plasma immunoreactive atrial natriuretic peptide (IR-ANP) was examined in anaesthetized rabbits, and the role of arterial pressure in mediating the changes in IR-ANP was assessed. Plasma IR-ANP was significantly greater (101.7 +/- 24.3 pg ml-1) when carotid sinus pressure was 60 mmHg than when it was 160 mmHg (27.1 +/- 8.6 pg ml 1). Mean arterial pressure (MAP) was significantly greater when carotid sinus pressure was controlled at 60 mmHg compared to when it was 160 mmHg, but right atrial pressure (RAP) was not significantly different at the two carotid sinus pressures. The administration of hexamethonium attenuated the changes in MAP and heart rate (HR) which occurred in response to alterations in carotid sinus pressure, and abolished the change in plasma IR-ANP. The results suggest that an inverse relationship exists between carotid sinus pressure and plasma IR-ANP, and that the release of ANP in response to a reduction of carotid sinus pressure is mediated by the associated haemodynamic changes. PMID- 2568571 TI - Dynorphin A (1-13), microinjected into the preoptic area, stimulates water intake in rats. AB - The effects of dynorphin A (1-13) (DYN), injected into the preoptic area, was investigated on water intake in rats. DYN at both doses of 2 and 10 nmoles significantly increased water intake for two and four hours after the injection in a dose related fashion. However, no significant change was observed in food intake. Naloxone pretreatment (0.3 mg/kg, s.c.) completely attenuated the DYN induced stimulation of water intake. The present studies suggest that DYN in the preoptic area may play an important role in the regulation of drinking behavior, but have no effect on food intake. PMID- 2568572 TI - Changes in expression of tyrosine hydroxylase immunoreactivity in human SMS-KCNR neuroblastoma following retinoic acid or phorbol ester-induced differentiation. AB - Human SMS-KCNR cells differentiated in response to either retinoic acid or phorbol esters; differentiated cells extended numerous, complex neurites and showed reduced proliferation. Tyrosine hydroxylase (TH) immunoreactivity was measured in this cell line following treatment with retinoic acid (1-10 microM), 12-O-tetradecanoyl-phorbol-13-acetate (TPA; 16-160 nM), or combinations of these agents. After 21 days of treatment with either TPA or retinoic acid (RA), TH immunoreactivity was measured in this using densitometric scans of Western blots, was doubled relative to untreated or serum-deprived SMS-KCNR cultures. Increases in TH immunoreactivity could be detected after 6 days of treatment. Treatment with RA for 3 days followed by phorbol esters for an additional 3 days resulted in a 3-fold increase in TH immunoreactivity at day 6; reversing the order of drug treatment did not have this effect. Treatment of cultures with the divalent cationophore A23187 caused treated cells to retract neurites; expression of TH immunoreactivity was decreased relative to drug-treated and control cultures. These results suggest that retinoic acid treatment may 'prime' SMS-KCNR cells for the subsequent effects of phorbol esters, and indicate that the patterns of biochemical differentiation induced by TPA or RA are different. PMID- 2568573 TI - Immigration, stress, and prescription drug use among Cuban women in south Florida. PMID- 2568574 TI - Characterization and molecular cloning of the PCF8775 CS5 antigen from an enterotoxigenic Escherichia coli 0115:H40 isolated in Central Australia. AB - The genes determining the biosynthesis of the colonization factor CS5 have been cloned from Escherichia coli 0115:H40:PCF8775 isolated during an outbreak of diarrhoea among aboriginal children in Central Australia. Electron microscopy has shown purified CS5 to be of semi-rigid fimbrial type. NH2-terminal analysis has shown the CS5 determinant to be distinct from other fimbriae, although there is some conservation of certain residues. Expression in minicells of the cloned fimbrial genes encoded on pPM1312 has shown that proteins of 70 and 46.5 kD which co-purity with the 23 kD major fimbrial subunit protein are also co-expressed along with proteins of 45, 31, 17 and 14 kD. The major CS5 subunit is synthesized in precursor form (approximately 26 kD). A synthetic oligonucleotide to the NH2 terminal amino acid coding sequence of the purified protein has been used in Southern hybridization analyses to define the region on pPM1312 encoding the structural gene for the major pilin subunit. PMID- 2568575 TI - The O75X adhesin of uropathogenic Escherichia coli is a type IV collagen-binding protein. AB - Interaction of the basement-membrane binding O75X adhesin of uropathogenic Escherichia coli with various extracellular matrix proteins was studied. The adhesin showed strong binding to type IV collagen immobilized on microtitre plates, whereas other collagens, laminin and fibronectin, were only weakly recognized. Similarly, specific binding of [125I]-labelled type IV collagen to O75X-positive bacteria was shown. Interaction of the two proteins was also demonstrated by affinity chromatography of the O75X adhesin on immobilized type IV collagen. The adhesin bound strongly to the immobilized N-terminal 7S domain of type IV collagen, and the binding of [125I]-labelled type IV collagen to O75X positive bacteria was inhibited by the soluble 7S domain. Binding of O75X to type IV collagen and to its 7S domain was specifically inhibited by chloramphenicol but was not affected by periodate or endoglycosidase-H treatment of the glycoproteins. Our results show that the 7S domain of type IV collagen is the basement membrane receptor for the O75X adhesin and suggest an interaction based on protein-protein recognition. Inhibition of the interaction by chloramphenicol favours the supposition that a modified tyrosine is involved in the binding site. PMID- 2568576 TI - The biochemical basis for growth inhibition by L-phenylalanine in Neisseria gonorrhoeae. AB - Clinical isolates of Neisseria gonorrhoeae are commonly subject to growth inhibition by phenylpyruvate or by L-phenylalanine. A blockade of tyrosine biosynthesis is indicated since inhibition is reversed by either L-tyrosine or 4 hydroxyphenylpyruvate. Phenylalanine-resistant (PheR) and phenylalanine-sensitive (PheS) isolates both have a single 3-deoxy-D-arabino-heptulosonate 7-phosphate (DAHP) synthase that is partially inhibited by L-phenylalanine (80%). However, PheS and PheR isolates differ in that the ratio of phenylpyruvate aminotransferase to 4-hydroxyphenylpyruvate aminotransferase is distinctly greater in PheS isolates than in PheR isolates. A mechanism for growth inhibition is proposed in which phenylalanine exerts two interactive effects. (i) Phenylalanine decreases precursor flow to 4-hydroxyphenylpyruvate through its controlling effect upon DAHP synthase; and (ii) phenylalanine is largely transaminated to phenylpyruvate, which saturates both aminotransferases, preventing transamination of an already limited supply of 4-hydroxyphenylpyruvate to L-tyrosine. PMID- 2568577 TI - Carteolol and penbutolol for hypertension. PMID- 2568578 TI - Linkage analysis of the myosin heavy chain gene in Dictyostelium discoideum using a mutation generated by homologous recombination. AB - A mutation (mhcA1 in strain HMM) created by insertional gene inactivation was used to map the Dictyostelium discoideum myosin heavy chain gene (mhcA) to linkage group IV. Three phenotypic traits associated with this mutation (slow colony growth, inability of the mutant to develop past aggregation, and the presence of five to ten integrated vector copies) cosegregated as expected for the consequences of a single insertional event. This linkage was confirmed using a restriction fragment length polymorphism. The mhcA1 mutation was recessive to wild type and was nonallelic with mutations at the following loci on linkage group IV: aggJ, aggL. couH, minA, phgB and tsgB. This work demonstrates the ability to apply standard techniques developed for D. discoideum parasexual genetic analyses to mutants generated by transformation, which is of particular relevance to analysis of genes for which no classical mutations or restriction fragment length polymorphisms are available. PMID- 2568579 TI - Delayed increase of Ca2+ influx elicited by glutamate: role in neuronal death. AB - The mechanism of delayed neurotoxicity, triggered by glutamate, was studied in 7 8-day-old primary cultures of rat cerebellar granule cells. Treatment of cultures for 15 min with 50 microM glutamate in Mg2+ -free medium, followed by removal of the excitoxin, resulted in neuronal death, which started to appear 2-3 hr after the termination of glutamate treatment. The number of dead neurons increased gradually in the next few hours and 80-85% of neurons were found dead 24 hr later. Antagonists of N-methyl-D-aspartate-sensitive glutamate receptors (phencyclidine) or 1.2 mM MgCl2, but not the antagonist of N-methyl-D-asparatate insensitive glutamate receptors (6-cyano-7-nitroquinoxaline-2,3-dione), abolished the neurotoxic effect of kainate. Development of glutamate-induced neuronal death depends strongly on Ca2+. Removal of extracellular Ca2+ (with 1mM ethyleneglycol bis-(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid) immediately after the termination of glutamate exposure and before the appearance of the early signs of neuronal death (post-glutamate period) dramatically reduced neuronal degeneration. Neurotoxic concentrations of glutamate induced sustained increase of 45Ca2+ uptake in the post-glutamate period. The delayed increase of 45Ca2+ uptake, as well as the delayed neurotoxicity, were not affected by post-glutamate treatment with phencyclidine, dibenzocyclohepteneimine; DL-2-amino-5 phosphonovalerate, or MgCl2 or with voltage-dependent Ca2+ channel blockers (nitrendipine, verapamil, diltiazem). Neurotoxic concentrations of glutamate also induced a delayed sustained increase of [3H]phorbol-12,13-dibutyrate binding, reflecting an increased translocation of protein kinase C (PKC) from cytosol to the cell membrane during the post-glutamate period. Pretreatment of neurons with the ganglioside GT1b (trisialosylgangliotetraglycosylceramide), followed by removal of free GT1b from the incubation medium, prevented PKC translocation, the sustained increase of 45Ca2+ uptake in the post-glutamate period, and the delayed neuronal death. We suggest that the sustained activation and translocation of PKC primed by glutamate receptor stimulation may be the triggering event causing the protracted increase of neuronal Ca2+ influx. This influx is insensitive to voltage-dependent Ca2+ channel blockers and glutamate receptor antagonists. It appears that this delayed increase of Ca2+ influx may be important in causing neuronal death. PMID- 2568580 TI - Tricyclic antidepressants and dextromethorphan bind with higher affinity to the phencyclidine receptor in the absence of magnesium and L-glutamate. AB - Recent studies from our laboratory have provided evidence that multiple states of the phencyclidine (PCP) receptor exist. In addition, several compounds such as PCP and the novel anticonvulsant MK-801 were found to inhibit binding more potently in the presence of Mg2+ and L-glutamate (L-GLU) than when these agents were excluded from the binding assay. In the present study, a number of pharmacological compounds that have been suggested to interact within the N methyl-D-aspartate (NMDA) receptor complex, including tricyclic antidepressants (TCAs), were examined for their ability to inhibit the binding of [3H]1-[1-(2 thienyl)cyclohexyl]piperidine [( 3H]TCP) in the absence or presence of Mg2+ and L GLU. The TCAs imipramine, amitriptyline, and opipramol produced shallow inhibition curves in the absence of Mg2+ and L-GLU. Computer analysis of the binding data indicated that a two-component binding model described the data significantly better than a one-component model. In the presence of Mg2+ and L GLU, the inhibition curves became steeper and were shifted to the right, and computer analysis of the binding data indicated that a one-component model adequately described the binding data. A series of other centrally active compounds, including several antipsychotics and antihistamines, the antiparkinsonian anticholinergic trihexyphenidyl and the antitussive dextromethorphan, were also found to be affected similarly by the inclusion of Mg2+ and L-GLU in the binding assay. Dextrorphan, in contrast to dextromethorphan, inhibited [3H]TCP binding more potently in the presence of Mg2+ and L-GLU. The present results suggest that the compounds that inhibit binding more potently in the absence of Mg2+ and L-GLU are interacting with the PCP receptor in a different manner from that of PCP and MK-801, because these open channel blockers inhibit [3H]TCP binding more potently in the presence of Mg2+ and L-GLU. The data support previous findings that TCAs interact with the NMDA receptor complex and suggest that the compounds trihexyphenidyl and dextromethorphan, which have been shown to block NMDA-mediated neurotoxicity, may produce their effects through an interaction with the PCP receptor, albeit by a different mechanism from that of open-channel blockers. PMID- 2568581 TI - Differential activity of a tissue-specific extinguisher locus in hepatic and nonhepatic cells. AB - Tissue-specific extinguisher 1 (Tse-1) is a genetic locus on mouse chromosome 11 that can repress expression of several liver genes in trans. This locus is clearly active in fibroblasts, as hepatoma cells retaining fibroblast chromosome 11 are extinguished for both tyrosine aminotransferase and phosphoenolpyruvate carboxykinase gene expression. To assess the activity of Tse-1 in other tissues, we transferred mouse chromosome 11 from several different cell types into rat hepatoma recipients. Tse-1 was active in nonhepatic cell lines derived from each primary germ layer, but Tse-1 activity was not apparent in hybrids between hepatoma cells and primary mouse hepatocytes. These differences in the genetic activity of murine Tse-1 were apparently heritable in cis. PMID- 2568582 TI - Expression of three stage-specific transcripts of AMP deaminase during myogenesis. AB - AMP deaminase, a ubiquitous enzyme in eucaryotes, plays a central role in energy metabolism. In the present study, RNase protection analyses and immunoprecipitation with tissue-specific antisera were used to examine the transcripts and peptides of AMP deaminase produced during myogenesis in vivo and during myocyte differentiation in vitro. In embryonic muscle and undifferentiated myoblasts, a 3.4-kilobase (kb) transcript encoded a 78-kilodalton (kDa) AMP deaminase peptide that cross-reacted with antisera raised to the AMP deaminase isoform purified from kidney of the adult animal. In perinatal muscle and myocytes at an intermediate stage of differentiation in vitro, a 2.5-kb transcript was produced, and it encoded a 77.5-kDa AMP deaminase peptide that cross-reacted with antisera to the isoform purified from adult heart muscle. At about the time of birth, another 2.5-kb AMP deaminase transcript that encoded an 80-kDa peptide became detectable. This peptide cross-reacted with antisera to the predominant isoform purified from adult skeletal muscle. PMID- 2568583 TI - Remarkable intron and exon sequence conservation in human and mouse homeobox Hox 1.3 genes. AB - A high degree of conservation exists between the Hox 1.3 homeobox genes of mice and humans. The two genes occupy the same relative positions in their respective Hox 1 gene clusters, they show extensive sequence similarities in their coding and noncoding portions, and both are transcribed into multiple transcripts of similar sizes. The predicted human Hox 1.3 protein differs from its murine counterpart in only 7 of 270 amino acids. The sequence similarity in the 250 base pairs upstream of the initiation codon is 98%, the similarity between the two introns, both 960 base pairs long, is 72%, and the similarity in the 3' noncoding region from termination codon to polyadenylation signal is 90%. Both mouse and human Hox 1.3 introns contain a sequence with homology to a mating-type controlled cis element of the yeast Ty1 transposon. DNA-binding studies with a recombinant mouse Hox 1.3 protein identified two binding sites in the intron, both of which were within the region of shared homology with this Ty1 cis element. PMID- 2568585 TI - 1,25-Dihydroxyvitamin D3 silences 3',5'-cyclic adenosine monophosphate enhancement of somatostatin gene transcription in human thyroid C cells. AB - Treatment of the somatostatin (SRIF)-producing TT cell line with 1,25 dihydroxyvitamin D3 (1,25 D3) lowered intracellular SRIF mRNA and peptide concentration. In separate experiments, the cAMP analog 8-(4-chlorophenylthio) cAMP stimulated a rapid increase in SRIF mRNA content of the TT cells and SRIF peptide secretion. To determine whether 1,25 D3 could inhibit either the transcriptional or secretory effects of cAMP, TT cells were pretreated with 1,25 D3 for 4 days followed by treatment with the cAMP analog. Pretreatment with 1,25 D3 inhibited the cAMP-mediated increase of SRIF mRNA, but had no effect on the secretory response. We conclude that the ability of 1,25 D3 to silence SRIF gene expression is more potent than cAMP enhancer activity but that 1,25 D3 has no effect on that portion of the cAMP-dependent pathway which regulates peptide secretion. PMID- 2568584 TI - Primary structure of a human mitochondrial protein homologous to the bacterial and plant chaperonins and to the 65-kilodalton mycobacterial antigen. AB - The complete cDNA for a human mitochondrial protein designated P1, which was previously identified as a microtubule-related protein, has been cloned and sequenced. The deduced amino acid sequence of P1 shows strong homology (40 to 50% identical residues and an additional 20% conservative replacements) to the 65 kilodalton major antigen of mycobacteria, to the GroEL protein of Escherichia coli, and to the ribulose 1,5-bisphosphate carboxylase-oxygenase (rubisco) subunit binding protein of plant chloroplasts. Similar to the case with the latter two proteins, which have been shown to act as chaperonins in the posttranslational assembly of oligomeric protein structures, it is suggested that P1 may play a similar role in mammalian cells. The observed high degree of homology between human P1 and mycobacterial antigen also suggests the possible involvement of this protein in certain autoimmune diseases. PMID- 2568586 TI - Clonality of parathyroid tumors in familial multiple endocrine neoplasia type 1. AB - Familial multiple endocrine neoplasia type 1 (MEN-1) is characterized by tumors of the parathyroids, endocrine pancreas, and anterior pituitary. Since the gene associated with MEN-1, located on chromosome 11 (11q13), may normally inhibit tumor proliferation, tumors could arise from inactivation of one or both of the alleles. However, parathyroid tumors in patients with MEN-1 have been considered to result from polyclonal hyperplasia. Using genetic probes, we tested parathyroid tumors for a monoclonal component, represented by a loss of alleles at any of eight loci along chromosome 11. Ten of 16 tumors from 14 patients with familial MEN-1 had losses of alleles from chromosome 11. Tumors with losses were larger than those without (1.6 vs. 0.2 g; P less than 0.002), suggesting that a monoclonal adenoma may develop after a phase of polyclonal hyperplasia. In 7 of 10 tumors, the subregion of loss was less than the full length of chromosome 11 but always included one copy of the MEN-1 locus. Of 34 sporadic adenomas from patients without MEN-1, 9 showed similar allelic losses in chromosome 11; in 7 the losses included the apparent MEN-1 locus. We conclude that many "hyperplastic" parathyroid tumors in familial MEN-1 are in fact monoclonal and may progress or even begin to develop by inactivation of the MEN-1 gene (at 11q13) in a precursor cell. Some sporadic adenomas have allelic losses on chromosome 11, which may also involve the MEN-1 gene. PMID- 2568588 TI - Preferential germline mutation of the paternal allele in retinoblastoma. AB - The event triggering malignant proliferation in 70% of retinoblastoma tumours is loss of heterozygosity for chromosome 13q14, whereby the normal retinoblastoma gene (RB1) allele is lost and an already mutated RB1 allele remains in the tumour. The first allele suffers a mutational event--deletion, duplication or point mutation (manuscript in preparation)--either in the germ line (all bilateral patients) or in a somatic retinal cell (most unilateral patients). Most bilateral patients have no family history of retinoblastoma and are presumed to have new germline mutations which arose in the egg, sperm or early embryo. We have determined the parental origin of the retained allele in nine retinoblastoma tumours from eight unrelated non-familial cases by using RB1-linked genetic markers. Six tumours retained the paternal allele and three retained the maternal allele. Of the three unilateral tumours, only one retained the paternal RB1 allele. Thus, there is no evidence that the paternal RB1 allele is preferentially retained in retinoblastoma, as has been suggested to be the case in osteosarcoma. By contrast, tumours from four of the five bilateral patients retained the paternal RB1 allele. This suggests either that new germline RB1 mutations arise more frequently during spermatogenesis than during oogenesis, or that imprinting in the early embryo affects chromosomal susceptibility to mutation. PMID- 2568587 TI - Association of parathyroid tumors in multiple endocrine neoplasia type 1 with loss of alleles on chromosome 11. AB - Familial multiple endocrine neoplasia type 1 (MEN-1) is an autosomal dominant disorder characterized by the combined occurrence of tumors of the parathyroid glands, the pancreas, and the pituitary gland. Pancreatic tumors have previously been shown to be associated with the loss of alleles on chromosome 11; we therefore looked for similar genetic alterations in specimens of parathyroid tumors, which are the most common feature of MEN-1. We obtained parathyroid tumors and peripheral-blood leukocytes from six patients with MEN-1; 18 cloned human DNA sequences from chromosome 11 were then used to identify restriction fragment-length polymorphisms. A loss of heterozygosity was detected in parathyroid tumors from three of the six patients with MEN-1; this finding demonstrated that allelic deletions on chromosome 11 are involved in the monoclonal development of parathyroid tumors in patients with MEN-1. In addition, studies of three affected families (with 17 affected members and 51 unaffected members) established linkage with the oncogene INT2 (peak lod score, 3.30, at 0 percent recombination); the MEN-1 gene was thus mapped to the pericentromeric region of the long arm of chromosome 11 (11q13). Our location of the MEN-1 gene at 11q13 is close to the location previously reported. We conclude that a single inherited locus on chromosome 11, band q13, causes MEN-1 and that the monoclonal development of parathyroid and pancreatic tumors in patients with MEN-1 involves similar allelic deletions on chromosome 11. PMID- 2568589 TI - [Pathophysiological bases and treatment of cardiac decompensation]. PMID- 2568591 TI - The effects of two anticonvulsants on amino acid levels in the developing rat cerebellum. AB - Two anticonvulsants were administered pre- and postnatally to determine their effects on putative amino acid neurotransmitter levels in the rat cerebellum. The amino acids were quantitated using precolumn fluorescence derivatization and reverse-phase high performance liquid chromatography at various postnatal intervals. Treatment with clonazepam produced an initial depression in levels of most of the amino acids analyised. By three weeks postnatal all the amino acids, with the exception of GABA, had returned to control levels. GABA levels were still depressed five weeks after the cessation of treatment. Phenobarbital treatment produced an initial elevation in the level of GABA. At three weeks postnatal, both GABA and glutamate levels were elevated and remained so at eight weeks postnatal. In conclusion, the data demonstrated that each anticonvulsant produced unique, acute and chronic alterations in the levels of the cerebellar amino acids. PMID- 2568590 TI - Effects of dorsal root section and occlusion of dorsal spinal artery on the neurotransmitter candidates in rat spinal cord. AB - In order to obtain further evidence of putative neurotransmitters in primary sensory neurons and interneurons in the dorsal spinal cord, we have studied the effects of unilateral section of dorsal roots and unilateral occlusion of the dorsal spinal artery on cholinergic enzyme activity and on selected amino acid levels in the spinal cord. One week after sectioning dorsal roots from caudal cervical (C7) to cranial thoracic (T2) levels, the specific activity of choline acetyltransferase (ChAT) was significantly decreased and acetylcholinesterase (AChE) showed a tendency to decrease in the dorsal quadrant on the operated side of the spinal cord. Dorsal root sectioning had little effect on the levels of free glutamic acid or other amino acids in the dorsal spinal cord. These results suggest that primary sensory neurons may include some cholinergic axons, and that levels of putative amino acid transmitters are not regulated by materials supplied by axonal transport from the dorsal root ganglia. By contrast, one week following unilateral occlusion of the dorsal spinal artery, the activities of ChAT and AChE were unchanged in the operated quadrant of the spinal cord, while decreases of Asp, Glu, and GABA, and an increase in Tau were detected. These findings are consistent with the proposals that such amino acids, but not ACh, may function as neurotransmitter candidates in interneurons of the dorsal spinal cord. PMID- 2568593 TI - Cytosolic-free calcium and neurotransmitter release with decreased availability of glucose or oxygen. AB - Exposing brain slices to reduced oxygen tensions or impairing their ability to utilize oxygen with KCN decreases acetylcholine (ACh) but increases dopamine (DA) and glutamate in the medium at the end of a release incubation. To determine if these changes are due to alterations in the presynaptic terminals, release from isolated nerve endings (i.e. synaptosomes) was determined during histotoxic hypoxia (KCN). KCN reduced potassium-stimulated synaptosomal ACh release and increased dopamine and glutamate release. Since several lines of evidence suggest that altered calcium homeostasis underlies these changes in release, the effects of reducing medium calcium concentrations from 2.3 to 0.1-mM were determined. In low calcium medium, KCN still increased dopamine and glutamate release, but had no effect on ACh release. Hypoxia increased cytosolic-free calcium in both the normal and low calcium medium, although the elevation was less in the low calcium medium. Thus, the effects of histotoxic hypoxia on cytosolic free calcium concentration paralleled those on glutamate and dopamine release. Reducing the glucose concentration of the medium also increased cytosolic-free calcium. The data are consistent with the hypothesis that hypoxia and hypoglycemia increase cytosolic-free calcium, which stimulates the release of dopamine and glutamate, whose excessive release may lead to subsequent cellular damage postsynaptically. PMID- 2568592 TI - A protein-free diet changes synaptosomal membrane fluidity and tyrosine and glutamate transport. AB - Synaptosomes were isolated from cerebrums of rats fed standard (20% protein) or protein-free diets for 30 days. Arrhenius plots of their (Na+/K+)ATPase activities revealed a transition temperature of 25.5 degrees C for control rats and 23.4 degrees C for rats on protein-free diet, indicating that the latter increases synaptosomal membrane fluidity. The only change observed in the composition of the synaptosomal membranes was a 26% decrease of sialic acid. In synaptosomes from rats on protein-free diet the uptake of tyrosine was slightly reduced while that of glutamate was not affected. However, the exit of glutamate was reduced. PMID- 2568595 TI - Responsiveness to dopaminergic agonists after devascularizing lesions of the cortex. AB - Unilateral devascularizing lesions of neocortex of the rat altered the responsiveness to the direct and indirect dopaminergic agonists, apomorphine and methamphetamine, respectively. These lesions produced subcortical damage manifesting itself as (1) decreased responsiveness to drug-induced locomotor activity, (2) the development of drug-induced rotational activity and (3) loss of body weight. Administration of the monosialoganglioside GM1 (i.c.v.; 5 mg/kg/day; 7 days) reversed these effects when the subjects were tested 10 and 30 days after surgery. The data are interpreted as resulting from damage to dopaminergic systems in the striatum. PMID- 2568594 TI - Changes in serotonin metabolism in the rat raphe magnus and cardiovascular modifications following systemic administration of clonidine and other central alpha 2-agonists: an in vivo voltammetry study. AB - By using the in vivo voltammetry, it was demonstrated that an injection of clonidine induced both cardiovascular modifications (hypotension and bradycardia) and a decrease in the level of 5-hydroxyindolacetic acid (5-HIAA) in the ventromedial B3 serotonergic (5-HT) cell bodies of the medulla oblongata of the rat. The cardiovascular effects of clonidine and of two other imidazolic compounds (detomidine and medetomidine) are likely to be related to their alpha 2 adrenoceptor agonist properties since hypotension and bradycardia were completely antagonized by idazoxan. The decrease in levels of 5-HIAA, induced by these three imidazolic compounds is likely to represent the combination of two additional mechanisms: (i) the stimulation of the alpha 2 adrenoceptors which could contribute to 55% of the decrease observed for the extracellular 5-HIAA and (ii) the interaction with a non-alpha 2 site (through a putative imidazole recognition site), corresponding to the part of the decrease (about 45%) which was not prevented by idazoxan. PMID- 2568596 TI - Agonist and antagonist properties of benzazepine and thienopyridine derivatives at the D1 dopamine receptor. AB - Nine structurally related 1-phenyl-1H-3-benzazepine derivatives and two thienopyridines were tested for agonist and antagonist properties at the adenylate cyclase-coupled D1 dopamine receptor in homogenates of the striatum of the rat. The benzazepines SK&F 77434 and SK&F 82958, both of which contain a catechol ring, were agonists; the intrinsic activity of SK&F 77434 was similar to that of SK&F 38393, whereas SK&F 82958 was a full agonist. The remaining benzazepines inhibited the stimulation of adenylate cyclase by dopamine. Antagonist potency depended on the nature of the substituent at position 7 of the benzazepine molecule, 7-halogen compounds being the most potent. The Ki values, obtained from analysis of the antagonism of dopamine-stimulated adenylate cyclase, were significantly correlated with the Ki values for displacement of D1 ligands in binding experiments. Furthermore, antagonist activity of the resolved racemic benzazepine SK&F 83566 resided almost exclusively in the R-enantiomer. The thienopyridine derivatives SK&F 89641 and SK&F 89145 were partial agonists with greater efficacies than SK&F 38393. PMID- 2568597 TI - Stimulation of central D1 dopamine receptors reverses reserpine-induced hypothermia in mice. AB - In mice rendered poikilothermic by a prior (18 h) subcutaneous administration of reserpine (3 mg/kg) the injection of the D1 dopamine agonist SKF 38393 in doses of 1 mg/kg or more increased dose-dependently, the body temperature. The D1 dopamine antagonist SCH 23390, administered subcutaneously, antagonized, with an ID50 of 16 micrograms/kg, the reversal by SKF 38393 of reserpine-induced hypothermia. The intracerebroventricular administration of 1 microgram per mouse of SKF 38393 was sufficient to elevate by about 7 degrees C the temperature of reserpinized mice. It is concluded that stimulation of central D1 dopamine receptors leads to a marked reversal of reserpine-induced hypothermia; this may constitute a new test to investigate interaction of drugs with these receptors. In reserpine-pretreated mice, the dopamine (DA) agonist apomorphine, which stimulates both the D1 and D2 subtypes of DA receptors, increases body temperature according to a mechanism insensitive to the specific D2 DA antagonist sulpiride (Horowski 1978) or the preferential D2 DA antagonist haloperidol (Danielson, Coutts, Keashly and Tang 1985). This observation led us to believe that D1 DA receptors could be involved in the reversal of the hypothermia induced by reserpine. To check more directly the involvement of D1 DA receptors in the reversal of the reserpine-induced hypothermia we have tested the specific D1 agonist SKF 38393 (Setler, Sarau, Zirckle and Saunders, 1978), administered peripherally or intracerebroventricularly and we have studied its interaction with the specific D1 antagonist SCH 23390 (Iorio, Barnett, Leitz, Houser and Korduba, 1983). PMID- 2568599 TI - Simultaneous use of digoxigenin- and radiolabeled oligodeoxyribonucleotide probes for hybridization histochemistry. AB - Oligodeoxyribonucleotide probes have proven very useful for studying gene expression in tissue sections, especially from the central nervous system. We report here a procedure for simultaneously identifying two different mRNA species in the same sections by hybridizing one with a digoxigenin-labeled probe, the other with a radiolabeled probe. The former probe is located by alkaline phosphatase-conjugated antibodies against digoxigenin, the latter by conventional nuclear emulsion autoradiography. Although the digoxigenin detection scheme currently lacks the sensitivity of autoradiography, it offers greater rapidity, resolution, and ease of detection of abundant transcripts. Furthermore, changes in gene expression within a subset of characterized cells is now feasible. PMID- 2568598 TI - Specificity of action of human brain alanyl aminopeptidase on Leu-enkephalin and dynorphin-related peptides. AB - The major cystosolic aminopeptidase (alanylaminopeptidase) was purified to homogeneity from human cerebral cortex and the specificity of its actions on a series of Leu-enkephalin-related peptides of increasing chain length was determined. In each case, only the N-terminal Tyr-Gly bond was hydrolysed. Kinetic analysis of the data revealed that the specificity constant (kcat/Km;s-1M 1) falls with increasing chain length from a maximum of 13.6 x 10(4) for Leu enkephalin (5 residues) to 5.8 x 10(2) for dynorphin (1-13). Dynorphin 1-17, while not being degraded itself acted as a competitive inhibitor (Ki = 2.7 microM) of the degradation of smaller peptides. Beta-endorphin was not hydrolysed by analylaminopeptidase, nor did it act as an inhibitor of the enzyme. PMID- 2568600 TI - Bestatin potentiates the antinociception but not the motor dysfunction induced by intracerebrally administered dynorphin-B in mice. AB - The intracerebrally administered dynorphin-B produced not only an antinociceptive response but a motor dysfunction such as "barrel-rolling", circling, jumping or ataxia in mice. These two effects were observed in a same dose range. Both responses were also produced by a nonopioid fragment, des-Tyr-dynorphin-B. Bestatin, an aminopeptidase inhibitor, markedly potentiated the antinociceptive response induced by dynorphin-B but not the motor dysfunction. Bestatin did not affect the responses produced by des-Tyr-dynorphin-B. In the presence of bestatin, low doses of dynorphin-B produced an antinociceptive response without the motor dysfunction. Naloxone antagonized the potentiated antinocicpetion but had no effect on the motor dysfunction. These results suggest that dynorphin-B produced an analgesia through opioid receptors and that this peptide also induced a motor dysfunction through a nonopioid receptor. PMID- 2568601 TI - 86Rubidium release from cultured primary astrocytes: effects of excitatory and inhibitory amino acids. AB - The effects of high K+, glutamate and its analogue, kainate, on K+ release were studied in primary astrocyte cultures prepared from newborn rat brains using 86Rb+ as a tracer for K+. An increase in 86Rb+ release was observed when the extracellular K+ concentration was elevated (10-40 mM). Glutamate and kainate stimulated the release in a dose-dependent manner, 100 microM concentrations being about as equally effective as high K+ (40 mM). Both compounds also caused an increase in the absorbance of the cyanine dye, 3,3'-diethylthiadicarbocyanine, indicating depolarization of the membrane. No significant Na+-dependent uptake of [3H]kainate occurred in the cells, thus excluding the possibility that depolarization was due to electrogenic uptake of amino acid into the cells. GABA and taurine significantly depressed the high K+- and glutamate-induced 86Rb+ release. Taurine itself caused a small increase in 86Rb+ release and the membrane was depolarized, judging from the increase in the absorbance of the cyanine dye, 3,3'-diethylthiadicarbocyanine. No effect of taurine was observed when the Cl- concentration was reduced in the experimental medium. The results suggest that cultured astrocytes respond by membrane depolarization to high external K+ and to glutamate and kainate. The degree of this depolarization can be modified by the inhibitory amino acids GABA, taurine and glycine, the effect of taurine probably being mediated by an increase in Cl- conductance across the cell membrane. The role of functional receptors for amino acid transmitters and the effects observed are discussed. PMID- 2568602 TI - Subchronic administration of MK-801 in the rat decreases cortical binding of [3H]D-AP5, suggesting down-regulation of the cortical N-methyl-D-aspartate receptors. AB - The effects of the subchronic administration of (+)-5-methyl-10,11-dihydro-5H dibenzo[a,d]-cyclohepten-5,10-imine (MK-801) (0.5 mg/kg twice daily, 7 days) on N methyl-D-aspartate, phencyclidine and sigma binding sites, behaviour and catecholamine turnover were investigated in the rat. Overt behaviours induced by MK-801 on day 7 were significantly altered relative to day 1 with subchronically treated rats not showing head weaving, goss ataxia or loss of hindlimb control: locomotion and sniffing were largely unaffected. The mean intensities of behaviour were 1.8 and 5.4 for days 7 and 1, respectively. Behavioural tolerance was accompanied by a significant reduction in the density of cortical N-methyl-D aspartate receptors as measured by [3H]D-2-amino-5-phosphonopentanoic acid binding, while affinity was unchanged: the density of binding sites was 3.52 and 1.88 pmol/mg protein for saline- and MK-801-treated rats, respectively. The N methyl-D-aspartate ion channel as measured by the binding of [3H]N-(1-[2 thienyl]cyclohexyl)piperidine was not affected by the schedule of MK-801. Additionally, changes were not observed to N-methyl-D-aspartate- or glycine stimulated [3H]N-(1-[2-thienyl]cyclohexyl)piperidine binding or to sigma binding. Catecholamine turnover was unaltered in the nucleus accumbens septi after the schedule of MK-801. Our results demonstrate that the subchronic administration of MK-801 produces behavioural tolerance and down-regulation of N-methyl-D-aspartate binding sites and suggest differential regulation of the domains of the N-methyl D-aspartate receptor-ionophore complex. PMID- 2568603 TI - Co-localization of glutamate and tubulin in putative excitatory neurons of the hippocampus and amygdala: an immunohistochemical study using monoclonal antibodies. AB - Tubulin and glutamate were immunohistochemically localized in the hippocampus and amygdala of rats using monoclonal antibodies to gamma-Glu-Glu (Glu-1) and glutaraldehyde-fixed glutamate (Glu-2), respectively. Glu-2 was shown to be selectively immunoreactive for glutaraldehyde-fixed Glu using enzyme-linked immunoassays and inhibition enzyme-linked immunoassays. Glu-1 was immunoreactive with tubulin on immunoblots of brain homogenates. However, only tubulin with a glutamate carboxy-terminal appeared to be immunoreactive with Glu-1 since tubulin from Chinese hamster ovary cells was not immunoreactive on immunoblots unless the tubulin was first treated with carboxypeptidase. Intense immunocytochemical staining by Glu-1 of hippocampus and amygdala was co-localized in the same neurons as the immunocytochemical staining for glutaraldehyde-fixed Glu produced by Glu-2. The distribution of immunostaining in the brain by Glu-1 was very similar to the distribution of immunostaining produced by Glu-2. The major difference was that glutamate-like immunoreactivity, visualized by Glu-2 staining, was intense in the nuclei of neurons, while nuclei were unstained by Glu-1. The distribution of immunostaining by these monoclonal antibodies was very similar to that reported in previous studies using other antibodies to Glu. All granule cells in the area dentata of the hippocampus exhibited intense immunoreactivity with both antibodies. Immunoreactivity was also observed in the stratum lucidum of CA3, the zone of termination of mossy fiber axons of granule cells. The majority of pyramidal cells in CA1, and many pyramidal cells in CA3 of the hippocampus were immunoreactive. In addition, it appeared that all of the pyramidal cells in the subiculum exhibited immunoreactivity. Light, diffuse immunoreactivity was observed in the neuropil of the hippocampus and subiculum. Most perikarya in the amygdala were characterized by light to moderate Glu-1 immunoreactivity and moderate to intense Glu-2 immunoreactivity. Fairly intense Glu-1 and Glu-2 immunoreactivity was seen in some neurons of the lateral nucleus, basolateral nucleus, lateral subdivision of the central nucleus, and the periamygdaloid cortex. The morphology of immunostained neurons in the lateral and basolateral nuclei indicates that the majority of these cells correspond to the pyramidal class 1 neurons described in previous Golgi studies.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2568604 TI - Immunocytochemical localization of thyroid hormone nuclear receptors in cultured hypothalamic dopaminergic neurons. AB - By means of a monoclonal antibody against the rat liver L-triiodothyronine nuclear receptor and a polyclonal anti-tyrosine hydroxylase serum, it has been possible to demonstrate thyroid hormone nuclear receptors in immunoreactive tyrosine hydroxylase cell nuclei in fetal rat hypothalamic cultures. After 8 days in vitro, the ratio of tyrosine hydroxylase cells that were immunoreactive for the thyroid hormone receptor to those not stained for this receptor (64% to 36% respectively) remains unchanged despite an increase in the number of tyrosine hydroxylase-positive cells with time (from day 8 to day 21) in culture. The presence of thyroid hormone nuclear receptor in dopaminergic neurons is correlated with a morphological effect of L-triiodothyronine in this neuronal population. Our results demonstrate, for the first time, the presence of triiodothyronine nuclear receptors in fetal rat dopaminergic neurons and the existence of a cellular heterogeneity in the distribution of the thyroid hormone receptor. The presence of these receptors in fetal hypothalamic dopaminergic neurons suggests that some effects of L-triiodothyronine on the maturation of DA neurons may result from a direct effect of this hormone through an interaction with its specific nuclear receptors. PMID- 2568605 TI - Quisqualate neurotoxicity: a delayed, CNQX-sensitive process triggered by a CNQX insensitive mechanism in young rat hippocampal slices. AB - Exposure of slices of young rat hippocampus for 30 min to the glutamate receptor agonist, quisqualate (QA, 30 microM), led, after a 90 min recovery period, to severe 'dark cell degeneration' of pyramidal neurones, most extensively those in CA3. When present during the exposure, 6-cyano-2,3-dihydroxy-7-nitroquinoxaline (CNQX, 10 microM), an antagonist with preferential action on non-N-methyl-D aspartate receptors, did not prevent this toxic effect of QA. However, it was effective when included either during the recovery period as well or, indeed, only during recovery. Comparable results were obtained with kynurenate (3 mM), but not with D,L-2-amino-5-phosphonovalerate (100 microM) or with tetrodotoxin (0.5 microM). Grease-gap recordings showed that CNQX markedly inhibited QA induced depolarization. It is concluded that QA toxicity is not triggered by QA induced depolarization but instead involves CNQX-resistant QA receptors, possibly those linked to phospholipid metabolism. The induction mechanism does not itself cause irreversible injury but subsequently, a delayed form of damage takes place which is mediated by activation of CNQX/kynurenate-sensitive receptors. PMID- 2568606 TI - Regulation of production by primary cultures of rat forebrain astrocytes of a trophic factor important for the development of cholinergic neurons. AB - The possibility was examined for the production of a trophic factor by astrocytes important for the biochemical development of cholinergic neurons. The activity of choline acetyltransferase (ChAT) was used as a marker to study the maturation of cholinergic neurons, while the metabolic state of astrocytes was monitored in terms of glutamine synthetase activity. When the dissociated cells, derived from the septal-diagonal band region of embryonic rat brain, were cultured either on a preformed layer of highly enriched astrocytes or in the presence of astrocyte conditioned medium, a marked increase was observed in the expression of ChAT activity. Similar effect was not noticed when cerebellar granule cells replaced the astrocytes. The production of cholinergic factor was decreased when the astroglial cells were induced to proliferate by epidermal growth factor, whereas its production was increased when quiescent astrocytes were treated with a low dose of cytosine arabinoside. The results would indicate that a trophic factor important for the development of cholinergic cells is produced by astrocytes, and that exogenously added agents influencing astroglial metabolism have a marked regulatory effect on the production of this trophic factor. PMID- 2568607 TI - Evidence for the existence of a population of arcuate neurons costoring choline acetyltransferase and tyrosine hydroxylase immunoreactivities in the male rat. AB - By combined immunoperoxidase and immunofluorescence histochemistry we have analyzed the distribution of choline acetyltransferase (ChAT) and tyrosine hydroxylase (TH) immunoreactive (IR) perikarya within the same sections of the mediobasal hypothalamus of the male rat. Evidence was obtained for the existence of perikarya costoring TH and ChAT immunoreactivities in both the dorsomedial and ventrolateral part of the arcuate nucleus and in the adjacent periarcuate nucleus at all rostrocaudal levels. The results strongly implicate interactions between dopamine and acetylcholine as well as acetylcholine and growth hormone releasing factor in dorsomedially and ventrolaterally located TH/ChAT costoring tuberoinfundibular neurons, respectively. PMID- 2568609 TI - Australian neuroscience proceedings. Abstracts of the ninth meeting of the Australian Neuroscience Society. Melbourne, Australia, February 6-8, 1989. PMID- 2568608 TI - Transient increase of NMDA-binding sites in human hippocampus during development. AB - In the human hippocampus, the density of glutamate and N-methyl-D-aspartate (NMDA) binding sites follows during development a bell-shaped curve with a peak at 23-27 fetal weeks. We suggest that there is a transient increased density of NMDA binding sites during a restricted period of hippocampal development; this may play an important role in developmental plasticity. PMID- 2568611 TI - Curriculum revolution: reconceptualizing nursing education. PMID- 2568610 TI - Inguinal and scrotal problems in infants and children. AB - Early and accurate diagnosis will not only prevent the catastrophe of gangrenous bowel in infants and children with inguinal and femoral hernia, but in the case of hydrocele will also avoid unnecessary parental anxiety. An accurate diagnosis and knowledge of the natural history of inguinal conditions can avoid unnecessary referral to surgical consultants. Finally, an understanding of the surgical expectations, ie, orchiopexy for the undescended testis, will provide parents and patients with realistic expectations. PMID- 2568612 TI - A new accI polymorphism for pMCT112 [D9S15]. PMID- 2568613 TI - XmnI, HincII and BclI RFLPs at D16S79. PMID- 2568614 TI - An AvaII polymorphism in the haptoglobin alpha gene (HPA). PMID- 2568616 TI - Ava-II RFLP at the human hepatic lipase (HL) gene locus. PMID- 2568615 TI - An HinfI polymorphism at the MET locus in inbred mice. PMID- 2568617 TI - L2, a DNA fragment from Xq24-q27, detects an EcoR1 RFLP (HGM9 no. DXS12). PMID- 2568618 TI - BamHI polymorphism at the human embryonic/atrial myosin alkali light chain gene locus. PMID- 2568619 TI - A new HLA-DQA1 RFLP allele [DQ alpha 3b] distinguishes between DQ alpha genes of DQw2-DR3 and DQw3-DR5 haplotypes. PMID- 2568620 TI - KpnI RFLP in the 3' flanking region of the C1 inhibitor gene. PMID- 2568621 TI - Taq I RFLP linked to the human ets-1 gene. PMID- 2568622 TI - Pushing for power. PMID- 2568623 TI - Opiate and alpha receptor antagonists block the pressor responses of conscious rats given intravenous dynorphin. AB - Conscious, unrestrained rats were used to determine the hemodynamic (blood pressure and heart rate) responses following intravenous (IV) injection of dynorphin A(1-13) and the possible receptor mechanisms mediating those changes. Male Sprague-Dawley rats (300 g) were given IV bolus injections (via femoral venous catheter) of 6.0 to 600 nmoles/kg of dynorphin A(1-13), 8.0 nmoles/kg of norepinephrine HCl (NE), 14.3 pmoles/kg of angiotensin II or a vehicle control solution. Blood pressure (BP) and heart rate (HR) were monitored via femoral arterial catheter (into abdominal aorta) over 90 sec postpeptide or -amine administration before and 10 min after IV injection of 4.2 mumoles/kg of naloxone HCl (opiate antagonist), yohimbine HCl (alpha 2 receptor antagonist) or prazosin HCl (alpha 1 receptor antagonist). Dynorphin A(1-13) caused a transient but dose related rise in mean arterial pressure (MAP) whereas mean pulse pressures (MPP) and mean heart rates (MHR) concomitantly fell, from preinjection control values in a dose-dependent fashion. Pretreatment with naloxone blocked the pressor response of only a subsequent injection with 20 nmoles/kg but not 60 nmoles/kg of dynorphin A or NE (8.0 nmoles/kg). Pretreatment with yohimbine suppressed the marked pressor responses of subsequent NE or Dyn A (60 nmoles/kg) administration whereas prazosin antagonized the rise in MAP of only the lower doses of dynorphin as well as NE. The suppression of the pressor responses of dynorphin by opiate or alpha receptor antagonists were not caused by tachyphylaxis for repeated injections of 6.0 or 60 nmoles/kg of dynorphin caused the same rise in MAP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568624 TI - MIF-1 attenuates spiroperidol alteration of striatal dopamine D2 receptor ontogeny. AB - Long-term postnatal treatment of rats with the dopamine D2 receptor antagonist, spiroperidol, results in the impaired development of striatal D2 receptors. Because the tripeptide prolyl-leucyl-glycinamide (MIF-1) attenuates haloperidol induced up-regulation of striatal dopamine D2 receptors in adult rats, we studied the effect of MIF-1 on the spiroperidol-induced alteration of striatal D2 ontogeny. Postnatal treatment of rats with spiroperidol (1.0 mg/kg/day, IP, x32 days from birth) resulted in a 74% decrease in the Bmax for [3H]spiroperidol binding with no change in the Kd at 5 weeks. When rats were studied at 8 weeks, in the absence of additional treatment, total specific [3H]spiroperidol binding was reduced by 59%. While MIF-1 alone (1.0 mg/kg/day, IP, x32 days from birth) had no effect on [3H]spiroperidol binding, MIF-1 completely attenuated the ontogenic impairment of striatal D2 receptors that was produced by spiroperidol treatment. At 5 weeks the Bmax for [3H]spiroperidol binding was at the saline control level in the group of rats cotreated with spiroperidol and MIF-1. At 8 weeks, with no additional treatments, the specific binding of [3H]spiroperidol to striatum was also at control levels in the group cotreated with spiroperidol and MIF-1. These findings demonstrate that MIF-1 attenuates spiroperidol-induced impairment of development of striatal dopamine D2 receptors in rats. PMID- 2568625 TI - Evidence for a single class of somatostatin receptors in ground squirrel cerebral cortex. AB - In the present study we characterized high-affinity somatostatin (SRIF) binding sites (Kd = 2.06 +/- 0.32 nM and Bmax = 295 +/- 28 fmol/mg protein) in cerebral cortex membrane preparations of European ground squirrel using 125I-[Tyr0-D-Trp8] SRIF14 as a radioligand. The inhibition of radioligand specific binding by SRIF14, as well as by its agonists (SRIF28, Tyr0-D-Trp8-SRIF14, SMS 201 995) was complete and monophasic, thus revealing a single population of somatostatinergic binding sites. Radioautographic analysis of 125I-[Tyr0-D-Trp8]-SRIF14 labeled brain sections confirmed the results of our biochemical study. The homogeneity of SRIF binding sites in the ground squirrel neocortex was not dependent on the animal's life-cycle phase. PMID- 2568626 TI - The involvement of opioid peptides in stress-induced analgesia in the slug Arion ater. AB - Application of tail-pinch stress to the terrestrial slug, Arion ater, produced a significant increase in the response time when tested on the hot-plate for foot lifting response. The analgesia was completely reversed by injections of the opiate antagonists, naltrexone and ICI 174864, in a dose-dependent manner. Analgesia could also be elicited by the injection into the foot of beta-endorphin and the enkephalin analogues, DAGO and DADLE. No effect was seen with dynorphin A (1-8) or dynorphin A (1-17). The stress-induced analgesia disappeared after 30 minutes but could be maintained for 100 min following the injection of a mixture of bestatin and the enkephalinase inhibitor, N-carboxymethyl-L-phenylalanyl-L leucine. This work suggests that in the slug, a physical stressor produces an analgesia which may be due to the release of endogenous opiates. PMID- 2568628 TI - Inhibitory effects of nasal drops components on granulocyte chemotaxis. AB - A possible toxic effect of components in nasal drops on chemotaxis by human granulocytes was studied. The vasoactive substances oxymetazoline chloride and xylometazoline chloride gave a successive reduction of chemotaxis down to zero for a concentration of 500 mg/l which is around that used in commercial preparations. The preservative benzalkonium chloride which is used in nasal drops in a concentration of 200 mg/l was deleterious for chemotaxis at a concentration of 0.8 mg/l. Thiomersal was deleterious for chemotaxis at a concentration of 1 mg/l which should be compared with a concentration of 24 mg/l used as preservative in nasal drops. Together with previous studies the present results indicate that the addition of preservatives in nasal drops should be questioned especially as they can be safely distributed without any risk of bacterial contaminations nowadays. PMID- 2568627 TI - Combined in situ hybridization and immunocytochemistry in the assay of pharmacological effects on tyrosine hydroxylase mRNA concentration. AB - An assay for tyrosine hydroxylase (TH) mRNA by in situ hybridization in combination with immunocytochemistry (ICC) for TH on the same section is described. The in situ hybridization protocol was optimized for [35S]cRNA (complementary RNA, i.e. anti-sense strand) probe concentration and time of hybridization. The specificity of hybridization was measured by several critera. The advantage of measuring grain density versus grains per cell is discussed for quantitation of in situ autoradiography. Finally, the reserpine-induced increase in adrenal TH mRNA was used to validate quantitative aspects of the in situ hybridization technique by comparison with blot hybridization. In contrast to the adrenal, reserpine did not increase TH mRNA in substantia nigra (s. nigra) neurons as measured by either technique. PMID- 2568629 TI - Attenuation of ischaemia-induced regional myocardial acidosis by bevantolol, a beta 1-adrenoceptor antagonist, in dogs. AB - In dogs anaesthetized with pentobarbital, the left anterior descending coronary artery (LAD) was occluded for 90 min. so that about 1/2 of the original flow was allowed to flow (partial occlusion). Bevantolol (a beta 1-adrenoceptor antagonist) or propranolol (a reference drug) was injected intravenously 30 min. after partial occlusion. Regional myocardial pH was measured by a micro glass pH electrode inserted in the LAD area. Partial occlusion decreased myocardial pH by 0.62 to 0.74. Bevantolol (1.0 mg/kg) or propranolol (1.0 mg/kg) significantly increased myocardial pH, that had been decreased by partial occlusion, within 60 min. after the injection. Restoration of myocardial [H+] (defined as return towards a lower [H+] to the preocclusion level) (calculated from the pH data) induced by bevantolol and that induced by propranolol were 64.0 and 66.4% (measured 60 min. after the injection), respectively. Bevantolol or propranolol decreased heart rate also. Even in the paced heart, bevantolol restored the myocardial [H+] that had been increased by partial occlusion. These results suggest that bevantolol has a favorable effect on the ischaemic myocardium as has propranolol, and that the pH effect of bevantolol is not primarily due to a decrease in heart rate. PMID- 2568630 TI - Dual effects of nicotine on neuroleptic-induced changes of striatal dopamine metabolism in mice. AB - The effect of nicotine on the neuroleptic-induced changes in striatal dopamine (DA) metabolism of mice was studied. To investigate the mechanism of action of nicotine, its interactions with apomorphine (APO) and gamma-hydroxybutyric acid (GHBA) were also investigated. Mice were given nicotine, (0.3-3 mg/kg subcutaneously) repeatedly (4 times) at 30 min. intervals. Haloperidol (HAL), (+/ )-sulpiride (SUL), APO or GHBA were administered after the second nicotine dose. Hexamethonium was given to prevent the effects of nicotine on autonomic ganglia. The striatal contents of DA and of its metabolites homovanillic acid (HVA), 3,4 dihydroxyphenylacetic acid (DOPAC) and 3-methoxytyramine (3-MT) were measured. The drug-induced hypothermia in mice was controlled by increasing ambient temperature. At ambient temperature of 32-34 degrees nicotine and HAL increased the striatal DOPAC and HVA contents additively, whereas APO counteracted the effects of nicotine at this ambient temperature. At 20-22 degrees nicotine decreased the 3-MT content which indicates reduced release of DA. In hypothermic mice nicotine inhibited better the HAL- and SUL-induced increases of HVA content than those of DOPAC content suggesting that the neuroleptic-induced increases in DOPAC and HVA contents are mediated partly by different mechanisms. In APO treated mice both the GHBA- and nicotine-induced decreases of 3-MT content fell further. GHBA did not alter the nicotine-induced decrease of 3-MT content and so this effect of nicotine may be mediated indirectly via GABAergic neurones. Unlike GHBA and APO nicotine decreased 3-MT content only in hypothermic mice.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568631 TI - Trimipramine and other antipsychotics inhibit Campylobacter pylori in vitro. AB - A series of conventional anti-ulcer drugs, tricyclic antidepressants and neuroleptics (and some CNS non-active isomers) were tested in vitro for possible inhibition of Campylobacter pylori. These bacteria are claimed to play an etiological role in peptic ulcer disease, at least in gastritis B. While cimetidine, famotidine, ranitidine and pirenzepine were inactive, all the antipsychotic agents and their isomeric derivatives were active to various degrees with IC50 of 26-59 microM. Of special interest is trimipramine (Surmontil) that has been demonstrated to be effective against duodenal ulcers in some trials. The activity of the non-neuroleptic stereo-isomers of clopenthixol and chlorprothixene may lead to investigation in patients with peptic ulcer disease of this kind of agents. However, a firm connection between the antimicrobial activity of these compounds, their possible anti-ulcer effect and the etiological role of Campylobacter pylori in peptic ulcer disease must first be established. PMID- 2568632 TI - Plasma concentrations of bromazepam following peroral and sublingual administration. PMID- 2568633 TI - Isolation of peroxisome-deficient mutants of Saccharomyces cerevisiae. AB - Two mutants of Saccharomyces cerevisiae affected in peroxisomal assembly (pas mutants) have been isolated and characterized. Each strain contains a single mutation that results in (i) the inability to grow on oleic acid, (ii) accumulation of peroxisomal matrix enzymes in the cytosol, and (iii) absence of detectable peroxisomes at the ultrastructural level. These lesions (pas1-1 and pas2) are shown to be nonallelic and recessive. Crossing of pas1-1 and pas2 strains resulted in diploid cells that had regained the ability to grow on oleic acid as sole carbon source and to form peroxisomes. These pas mutants may provide useful tools for future studies on the molecular mechanisms involved in peroxisomal assembly. PMID- 2568634 TI - Two steps in the evolution of Antennapedia-class vertebrate homeobox genes. AB - Antennapedia-class vertebrate homeobox genes have been classified with regard to their chromosomal locations and nucleotide sequence similarities within the 183 base-pair homeobox domain. The results of these comparisons support the view that in mammals and most likely the vertebrates, four clusters of homeobox genes exist that were created by duplications of an entire primordial gene cluster. We present evidence that this primordial cluster arose by local gene duplications of homeoboxes that were present before the divergence of arthropods and chordates. Sequence analyses indicate that the expansion of the primordial gene cluster complex was accompanied by diversification, whereas conservation predominated after the duplications of entire homeobox gene clusters. PMID- 2568636 TI - The homeotic gene spalt (sal) evolved during Drosophila speciation. AB - The region-specific homeotic gene spalt (sal) acts in two separate domains in the head and tail region of the Drosophila melanogaster embryo. Based on comparative morphology, sal is likely to be involved in the establishment of the head during the evolution of invertebrates and thus, it should be conserved. We have analyzed the conservation of the segmentation genes Kruppel (Kr) and even-skipped (eve) in parallel with sal coding sequences in several Drosophila species that are evolutionarily separated by up to 60 million years. To our surprise, sal sequences appear to be conserved in the Sophophora subgenus of the Drosophila genus but not in the Drosophila subgenus. On the other hand, the segmentation and other homeotic genes are conserved in the Drosophila subgroup as well. Our data suggest that sal encodes an accessory function that evolved relatively late during Drosophila speciation rather than playing a fundamental evolutionary role similar to that of other homeotic genes. PMID- 2568637 TI - Ultra-high resolution scanning electron microscopy of biological materials. PMID- 2568635 TI - Multiplex gene regulation: a two-tiered approach to transgene regulation in transgenic mice. AB - Transgenic mice have been used to study gene function and regulation by introducing inducible or tissue-specific transgenes. This approach is generally limited to studying gene function in adult mice since ectopic expression of many interesting genes is disease causing or may be lethal to the developing embryo. To extend the utility of the transgenic mouse system to the early stages of embryogenesis, we have developed a two-tiered method of gene regulation to control transgene expression. Our multiplex gene regulatory system (MGR) allows the establishment of transgenic lines that harbor inducible potentially lethal transgenes. These inducible transgenes are activated only when mated to a second transgenic animal. Induction in the MGR system provides a high degree of temporal and spatial control over transgene expression and should be suitable for engineering "gain of function mutations" for many developmental genes. PMID- 2568638 TI - Diversity in the chemical nature and mechanism of response to tumor promoters. PMID- 2568639 TI - Zuclopenthixol and haloperidol/levomepromazine in the treatment of elderly patients with symptoms of aggressiveness and agitation: a double-blind, multi centre study. AB - A double-blind study was carried out in 48 hospitalized, elderly demented patients with key symptoms of aggressiveness and agitation to evaluate the efficacy and tolerability of zuclopenthixol compared with that of haloperidol/levomepromazine. Patients were allocated at random to receive initial doses of either 4 mg zuclopenthixol daily or 1 mg haloperidol in the morning and 5 mg levomepromazine in the evening over a period of 4 weeks. In Week 4, the mean daily dose was 4.8 mg zuclopenthixol and 1.6/7.6 mg haloperidol/levomepromazine, respectively. After 1 week, the severity of illness was already significantly reduced, and was further reduced after 2 and 4 weeks of treatment in both groups: the reduction, however, was most pronounced in the zuclopenthixol group and after 2 weeks this difference was significant. Side-effects were few. The results of the study indicate that, whilst both zuclopenthixol and haloperidol/levomepromazine were effective and well tolerated in these elderly patients with aggressiveness and agitation, onset of therapeutic effect appeared more rapidly with zuclopenthixol, which furthermore provides the practical advantage that it may be administered once a day. PMID- 2568640 TI - Synthesis of some 2-(3,4-dimethoxy-phenyl)-1,3-propanediamines as potential dopaminergic agents. AB - The bilateral diamino analogs namely N,N'-dialkyl-N,N'-diaralkyl- and N,N,N',N' tetraalkyl-2-(3,4-dimethoxyphenyl)-1,3-propanediamine s were synthesized. The biochemical determination of the brain levels of dopamine and norepinephrine as well as of brain monoamine oxidase (MAO) activity was performed. PMID- 2568641 TI - [Biotransformation of B 24/76 in the rat, minipig, dog and man]. AB - After oral administration of B 24/76 (1) on experimental animals (rat, minipig, dog) and human beings 11 metabolites could be isolated from urine, faeces and bile. The structure of 7 compounds was elucidated by means of TLC and MS. The investigated species are showing an analogues metabolic pattern with differences in the intensity and main metabolites. In phase II-reaction we could observe qualitative differences. PMID- 2568642 TI - Monitoring of serum bioactivity levels of perazine and its metabolites by radioreceptor assay. AB - The bioactivity of perazine and its metabolites in human serum was analysed by radioreceptor assay. The IC50 value (the concentration eliciting a 50% blockade of 3H-spiroperidol binding to the dopamine receptor in a membrane suspension of porcine striatum) was for perazine 175 nmol/l, and for its metabolites perazine sulfoxide 1050 nmol/l, desmethylperazine 330 nmol/l, N-(3-phenothiazin-10-yl propyl)-ethylenediamine 2800 nmol/l, and N-(3-phenothiazin-10-yl-propyl)-N' methylethylenediamin 2850 nmol/l. Thus, perazine metabolites possess low affinity to the dopamine receptor. In a pilot study we measured the bioactivity of perazine and its metabolites in sera of perazine-treated psychiatric patients by radioreceptor assay and compared the results to levels obtained by high performance thin-layer chromatography; the correlation coefficient r was 0.85 and the slope 0.95 (n = 11). Thus serum perazine can be adequately monitored by radioreceptor assay, as evidenced by the results from high-performance thin-layer chromatography. PMID- 2568643 TI - A pilot study of the role of prophylactic antiparkinson treatment during neuroleptic therapy. AB - The authors administered haloperidol 4.5 mg t.i.d. to 33 drug-free schizophrenic patients. Ten patients did not receive anything else (group HPL), while ten patients received procyclidine 5 mg t.i.d., and 13 patients were given promethazine 25 mg t.i.d. (groups HPRC and HPRM respectively) in addition. Seven patients dropped out of the HPL group and three out of the HPRM group, but none out of the HPRC group. These drop outs were due to the development of early extrapyramidal side effects, which were absent in the HPRC group. The findings suggest that antiparkinson prophylaxis is useful during commencement of therapy with high-potency neuroleptic agents. PMID- 2568644 TI - Benzodiazepines: utilization and patterns of use in a university hospital. AB - Information on the number of benzodiazepine prescriptions in all the departments of the Innsbruck University Clinics in 1985 was obtained from the hospital pharmacy. Expenditure on benzodiazepines amounted to 0.5% of total medication costs. In a second step, the authors questioned 264 patients from the five largest departments about benzodiazepine use. On the day of the interview, 24.1% of all investigated male patients and 20.5% of the female patients were taking benzodiazepines. Of the patients found to take benzodiazepines, 34.5% had already been using these before hospital admission. One-fifth of these patients reported problems when trying to stop medication. PMID- 2568645 TI - Double-blind study of metaclazepam versus diazepam treatment of outpatients with anxiety syndrome. AB - The therapeutic efficacy and tolerance of metaclazepam and diazepam were compared in a double-blind study of outpatients suffering from a generalized anxiety syndrome. The investigators were general practitioners. A total of 168 male and female patients aged between 18 and 60 years were included in the study and received either 15 mg metaclazepam or 15 mg diazepam per day. The analysis of tolerance was made for all 168 included patients, the evaluation of efficacy is based on the results of 131 patients (42 males and 89 females) with valid data over four weeks. During the four-week therapy period four examinations were made on days 0, 7, 14, and 28. A significant improvement of the severity of illness after administering the drugs was found for both drugs in the Clinical Global Impressions (CGI), in the Hamilton Anxiety Scale (HAMA), in the List of Complaints (B-L), and in the Adjective Checklist (EWL-K). Metaclazepam showed a statistically significant superiority over diazepam as far as the CGI items "severity of illness" and "global improvement" were concerned. Metaclazepam was slightly superior to diazepam in the two HAMA subscales "psychic anxiety" and "somatic anxiety". In the items of the selfrating scales (B-L and EWL-K) the therapeutic results of the metaclazepam group were, almost without exception, better than those of the diazepam group. A comparison of tolerance showed that metaclazepam was better tolerated. This can be seen in the greater frequency of side effects like tiredness and drowsiness under diazepam. Especially at the beginning of treatment, tiredness and drowsiness were recorded 2 1/2 times more frequently for the patients on diazepam than for those on metaclazepam. PMID- 2568646 TI - Neuroleptic-induced persistent "open mouth". AB - After prolonged exposure to butyrophenones, a patient with a paranoid psychosis showed an extrapyramidal motor disturbance which, phenomenologically, was very unusual and outlasted the medication. He involuntarily kept his mouth wide open all the time. Reasons for that symptom other than an induction by neuroleptics were not found. PMID- 2568647 TI - The cell division cycle of Trypanosoma brucei brucei: timing of event markers and cytoskeletal modulations. PMID- 2568648 TI - Modulation of neurotransmitter release by presynaptic autoreceptors. PMID- 2568649 TI - Management of the behavioral symptoms associated with dementia. AB - Dementia-related behavioral symptoms present a difficult management problem for caregivers and health care providers. The first step in the treatment of behavioral symptoms in the elderly should involve nonpharmacologic measures. This should include the exclusion of medication-induced dementia and delirium, which is an important consideration in the elderly owing to multiple medication use and alterations in pharmacokinetics and pharmacodynamics. Other measures involve relatively simple environmental changes. If these are unsuccessful, pharmacologic management is often tried. Numerous concerns surround the use of antipsychotics, which are the primary agents used to treat behavioral symptoms in demented elderly. The public is concerned about their inappropriate use as chemical restraints. Another concern involves the efficacy of these agents. Most well designed studies demonstrate an overall modest effect from the antipsychotics that is often similar to that seen with placebo. There may be a small number of patients who respond markedly, whereas the majority do not receive great benefit. Unfortunately, all are exposed to the numerous side effects of these agents, including some irreversible movement disorders. If these agents are believed to be necessary, efforts should be made to use them in a manner that reduces the associated risks. A few other agents have been noted to successfully treat the problem behaviors associated with dementia, but most have been in the form of case reports. It appears that the optimal treatment of these problems relies on future research in the area of nonantipsychotic modalities that will be efficacious and have a rather benign side effect profile. PMID- 2568650 TI - Tardive dyskinesia in schizophrenic patients: correlation with negative symptoms. AB - The relationship between severity of tardive dyskinesia (TD) and the prominence of negative symptoms was assessed in 25 right-handed, medicated schizophrenic patients. TD was quantified using ultrasound detectors and frequency measurement techniques as well as with observer rating scales. Electromechanical studies revealed a systematic relationship between TD severity and negative symptoms; TD was more severe in patients with fewer negative symptoms. The correlation was small in magnitude. PMID- 2568651 TI - Clinical implications of increased antipsychotic plasma concentrations upon anticonvulsant cessation. AB - Plasma antipsychotic concentrations were measured in five patients when their anticonvulsant medications were discontinued. Plasma antipsychotic concentrations increased by two- to five-fold at 4 weeks after cessation. All patients demonstrated either moderate or marked improvement as antipsychotic plasma concentrations increased. Four patients experienced extrapyramidal side effects within 30 days of anticonvulsant discontinuation. Careful monitoring of clinical symptoms, adverse effects, and plasma antipsychotic concentrations is recommended. PMID- 2568652 TI - Hallucinogenic and stimulatory amphetamine derivatives: fingerprinting DOM, DOI, DOB, MDMA, and MBDB by spectral analysis of brain field potentials in the freely moving rat (Tele-Stereo-EEG). AB - Telemetric recordings of field potentials from frontal cortex, hippocampus, striatum and reticular formation of freely moving rats were analysed before and after injection of the enantiomeric hallucinogenic amphetamine derivatives R-DOB [(-)-1-(2,5-dimethoxy-4-bromophenyl)-2-aminopropane], R-DOM [(-)-1-(2,5-dimethoxy 4-methylphenyl)-2-amino-propane] and R-DOI [(-)-1-(2,5-dimethoxy-4-iodophenyl)-2 aminopropane] as well as the nonhallucinogenic amphetamine derivatives S-MBDB [(+)-N-methyl-1-(1,3-benzodioxol-5-yl)butanamine] and S-MDMA [(+)-3,4 methylenedioxymethamphetamine] and S-(+)-amphetamine. The frequency analysis of the field potentials revealed a clearcut difference between them. The spectral patterns emerging after injection of the non-hallucinogens were characterized by a general decrease of power, the changes in the alpha2 and delta band being the most prominent, whereas only after the application of the hallucinogenic compounds was a contrasting increase of power observed in the alpha 1 frequency band, especially in the striatum. As increases in alpha 1 power have been correlated in the same pharmacological model to serotonergic control mechanisms, the results are in line with the hypothesis that 5-HT2 receptors, predominantly occurring in the striatum, might be involved in the hallucinogenic action of drugs. PMID- 2568653 TI - Remoxipride--a new potential antipsychotic compound. Tolerability and pharmacokinetics after single oral and intravenous administration in healthy male volunteers. AB - The tolerability and pharmacokinetics of remoxipride were studied in 18 healthy normal male volunteers. Increasing oral doses of 0.5-100 mg were given to eight male volunteers in one study (study I). In addition, an intravenous (IV) infusion of 20 mg remoxipride and a 20 mg oral dose were given in an open crossover study to ten males (study II). Remoxipride was well tolerated with respect to cardiovascular effects, clinical chemistry, body temperature and adverse effects in all subjects. Following IV administration, remoxipride plasma concentrations declined exponentially in five subjects and biexponentially in the remaining five. The mean apparent volume of distribution was 0.5 l/kg (SD = 0.10) and the mean half-life 4.1 h (range 2.6-6.6). The recovery of unchanged remoxipride in urine was 10-36%, and the mean renal clearance was 32 ml/min (SD = 13). Remoxipride was a low clearance drug with a total plasma clearance of about 120 ml/min (SD = 41). The mean oral bioavailability was 96%. There was a linear relationship between the peak plasma concentration as well as the area under the concentration versus time curve and the administered dose. A transient increase in plasma prolactin concentrations occurred but there were no effects on plasma growth hormone levels. PMID- 2568654 TI - Influence of the D-2 dopamine receptor agonist quinpirole on the odor detection performance of rats before and after spiperone administration. AB - The influence of five doses of the D-2 receptor agonist quinpirole (0.025, 0.05, 0.075, 0.10, and 0.20 mg/kg IP) on the odor detection performance of 21 adult male Long Evans rats was assessed using high precision olfactometry and a go/no go operant task. Additionally, ten rats were pre-treated with the D-2 receptor antagonist spiperone (0.62 mg/kg IP) and their performance monitored following quinpirole administration. Treatments were administered every third day in a counterbalanced order, with the quinpirole injections occurring 15 min before, and the spiperone injections 35 min before, the 260-trial test sessions. Quinpirole injection resulted in a dose-dependent decrease in odor detection performance, as measured by the percentage of correct trials and by the non parametric signal detection sensitivity index SI. Prior treatment with spiperone eliminated these effects. Dose-related influences of quinpirole on (a) the average latency to initiate a detection response (i.e., the S + response latency), (b) the total session duration, and (c) the number of aborted trials were also eliminated or greatly attenuated by prior spiperone injection. These results suggest that D-2 receptors may be involved in the modulation of odor detection performance and related behaviors. PMID- 2568655 TI - Synergistic blockade of some dopamine-mediated behaviours by (-)-sulpiride and SCH 23390 in the rat. AB - Several studies have indicated that the D2 dopamine receptors mediate the antidopaminergic activity of the neuroleptics; nevertheless, the selective blocker (-)-sulpiride weakly inhibits dopamine-mediated behaviour. The present study investigated whether the concomitant injection of doses of the D1 antagonist SCH 23390, which by themselves are without effect, would enable (-) sulpiride to express fully neuroleptic activity in the rat. The benzamide YM 09151-2 that strongly inhibits dopamine-mediated behaviour was also studied. Rats receiving different doses of (-)-sulpiride, YM 09151-2 and SCH 23390 given alone or in combination were tested for exploratory activity, apomorphine-induced stereotyped behaviour and hyperactivity elicited by the D2 agonist LY 171555. When given alone, (-)-sulpiride (10, 20 and 40 mg/kg IP) had no effect on exploratory activity and stereotypy. When (-)-sulpiride was administered in combination with an ineffective dose of SCH 23390 (5 micrograms/kg) both responses were significantly inhibited. The combined administration of subthreshold doses of (-)-sulpiride (2.5 mg/kg) and SCH 23390 (2.5 micrograms/kg) significantly inhibited hypermotility induced by LY 171555. Moreover, the combined administration of ineffective doses of YM 09151-2 with subthreshold doses of SCH 23390 strongly inhibited all the behavioural responses. The results indicate that SCH 23390 allowed (-)-sulpiride to exhibit a wider spectrum of neuroleptic activity and potentiated the antidopaminergic activity of YM 09151-2. PMID- 2568656 TI - Differential effects of dopamine receptor antagonists on the sexual behavior of male rats. AB - In the present experiments, the dose-response effects of the dopamine (DA) receptor antagonists haloperidol, pimozide, clozapine, sulpiride, and metoclopramide, were assessed on patterns of copulatory behavior in intact, sexually active male rats with a high level of sexual experience and performance. The typical neuroleptics haloperidol (0.01-0.5 mg/kg) and pimozide (0.1-5.0 mg/kg) dose-dependently delayed the initiation of copulation and reduced the number of intromissions that preceded ejaculation. The atypical neuroleptics clozapine (0.1-5.0 mg/kg), and sulpiride (0.1-5.0 mg/kg) dose-dependently delayed the initiation of copulation but had no effect on copulatory behavior once it was initiated. In contrast, metoclopramide dose-dependently reduced ejaculation but had no effect on the ability of rats to initiate copulation. These experiments suggest that aspects of copulatory behavior in male rats are affected differently by DA antagonists depending upon their site of action in the brain. Blockade of mesolimbic DA receptors by typical and atypical neuroleptics may delay the initiation of copulation, whereas blockade of mesostriatal DA receptors by typical neuroleptics and metoclopramide may decrease the ejaculation threshold. PMID- 2568657 TI - Chronic neuroleptic-induced mouth movements in the rat: suppression by CCK and selective dopamine D1 and D2 receptor antagonists. AB - Fluphenazine decanoate (25 mg/kg IM every 3 weeks x 6) resulted in spontaneous vacuous chewing mouth movements and jaw tremor in male Sprague-Dawley rats. These movements could be suppressed by the selective D1 or D2 dopamine antagonists SCH 23390 (0.5 mg/kg) and raclopride (0.5 mg/kg), respectively, and by CCK-8S (50 micrograms/kg). Fluphenazine-induced mouth movements were unaffected by the selective CCK antagonist MK-329, and by a dose of physostigmine (50 micrograms/kg) sufficient to stimulate mouth movements in placebo treated rats. Scopolamine (0.1 mg/kg) suppressed spontaneous mouth movements in placebo-treated rats, but the effect on fluphenazine-induced mouth movements was not significant. A higher dose of scopolamine (0.5 mg/kg) did suppress the neuroleptic-induced mouth movements, but also induced hyperactivity, characterized by increased sniffing and grooming. These findings indicate that mouth movements resulting from the chronic administration of neuroleptics to the rat may serve as a useful pharmacological model of tardive dyskinesia in the human, and suggest that a relative increase of D1 activity as well as impaired CCK function may contribute to the pathogenesis of this disorder. PMID- 2568658 TI - Effect of neurotransmitters on the system that transports Tyr-MIF-1 and the enkephalins across the blood-brain barrier: a dominant role for serotonin. AB - Neurotransmitters and neuropeptides interact in several ways. We studied a new type of interaction: the effect of neurotransmitters on the saturable system that transports Tyr-MIF-1 and the enkephalins out of the central nervous system (CNS). The neurotransmitters were introduced into the lateral ventricle of the brain with radioiodinated peptide, using an established method previously shown to accurately quantify the amount of peptide being transported from the CNS to the blood. Serotonin inhibited transport, histamine stimulated transport, and dopamine, acetylcholine, epinephrine, GABA, kainic acid, cAMP and cGMP were without effect. Cyproheptadine, a serotonin antagonist, stimulated transport. Of several psychotropic agents tested, only tranylcypromine had a statistically significant effect and stimulated transport. Of the serotonin receptor specific agents tested, those with 5HT1 activity most consistently affected transport. We conclude that serotonin, and perhaps histamine, are important modulators of the system that transports Tyr-MIF-1 and the enkephalins out of the CNS. PMID- 2568659 TI - Regional differences in the induction of behavioral supersensitivity by prolonged treatment with atypical neuroleptics. AB - Some atypical neuroleptics have been shown to exert selective effects on the nigrostriatal or mesolimbic dopamine systems as assessed by behavioral, biochemical and electrophysiological measures. This specificity appears to occur using chronic or acute schedules of drug administration. This study examined the effect of chronic administration of haloperidol, clozapine, sulpiride and metoclopramide on stereotypy and locomotor activity elicited by direct injection of dopamine in to the striatum or nucleus accumbens, respectively. Each rat was pre-treated with a neuroleptic drug or vehicle control for 21 days. Five days after the termination of drug treatment, each rat was injected with 10 micrograms dopamine bilaterally, and stereotypy or locomotor activity was measured. Rats pre treated with metoclopramide exhibited an enhanced stereotypy response, and rats pre-treated with haloperidol, clozapine or sulpiride exhibited enhanced locomotor activity compared to controls. The extent to which these results demonstrate the selective action of these drugs in sensitizing dopamine systems is discussed. PMID- 2568661 TI - Comparison of therapeutic efficacy of neuroses between CM6912 (ethyl loflazepate) and diazepam in a double-blind trial. AB - 1. The therapeutic efficacy on several neuroses between CM6912 and diazepam (DZP) was comparatively investigated by means of intergroup comparison method in a double-blind trials. CM6912 was given to CM-1 group on the basis of twice daily dose of 1 mg each in the morning and evening and to CM-2 group on the basis of once daily dose of 2 mg only in the evening and DZP was given to another group on the basis of three times each dose of 2 mg. 2. As for the overall evaluation, no significant differences were found in the percentages of final global improvement rating among three groups. It resulted in "markedly improved" by 22% in CM-2 group, 18% in CM-1 group and 15% in DZP group in order, and "more than moderately improved" by 62% in CM-2 group, 56% in DZP group and 51% in CM-1 group in order. 3. As for the overall safety rating, no differences were found in the incidence of adverse reactions. 4. As for the global utility rating, it resulted in "extremely useful" by 22% in CM-2 group, 14% in CM-1 group and 8% in DZP group in order, showing significant superiority of CM-2 group to DZP group (p less than 0.05). It resulted in "more than useful" by 58% in CM-2 group, 52% in DZP group and 50% in CM-1 group in order. PMID- 2568660 TI - Effects of a sedative and of a non-sedative H1-antihistamine on the event-related potential (ERP) in normal volunteers. AB - Measurements of the amplitude and latency of the P3b component of the event related potential (ERP), simple reaction time (SRT) and four psychomotor tests (VAS, DSST, DSp and CFF) were made on 12 male subjects (aged 19-24 years) 1.0-1.5 and 4.0-4.5 h after single oral doses of triprolidine (7.5 mg), terfenadine (60 mg) and placebo. Neither triprolidine nor terfenadine changed P3b amplitude or latency although VAS, CFF and DSST scores were significantly altered by triprolidine at 1.0-1.5 h after dosage. These results suggest that the P3b is too robust to reflect the mild sedative properties of an H1-receptor antihistamine, or that H1-receptors are not involved in P3b generation. PMID- 2568662 TI - Pharmacokinetics of three different injectable zuclopenthixol preparations. AB - 1. The zuclopenthixol concentrations in serum has been investigated in man and dog after injection of three different zuclopenthixol preparations. These were zuclopenthixol dihydrochloride in aqueous solution, zuclopenthixol acetate in oil and zuclopenthixol decanoate in oil. 2. The pharmacokinetic profiles of the three injectable zuclopenthixol preparations are very different. Maximum serum levels are obtained after about 1 hour for zuclopenthixol dihydrochloride, after 36 hours for zuclopenthixol acetate and after one week for zuclopenthixol decanoate. 3. The different pharmacokinetics of the three injectable zuclopenthixol preparations are reflected in their clinical properties. PMID- 2568663 TI - Influence of CGP 361A, propranolol and diazepam on autonomous reactions to different stressors. AB - 1. To evaluate the influence of beta-blockers and minor tranquilizers on autonomous stress response, 40 healthy subjects selected with regard to personality traits were randomly assigned to 5 groups. 2. Mental arithmetic induced the most pronounced increase in heart rate (HR), blood pressure and epinephrine secretion. 3. Beta-blockers reduced HR increases due to mental stress, whereas the minor tranquilizer reduced skin conductance level throughout the whole trial. 4. Our data provide evidence that different structured situations induce specific response patterns that are differentially modified by beta-blockers and minor tranquilizer. PMID- 2568664 TI - Brain monoamine oxidase (MAO) B: a unique neurotoxin and neurotransmitter producing enzyme. AB - The notion that monoamine oxidase (MAO) functions solely to inactivate neurotransmitter and other biogenic amines needs to be re-evaluated. It is now apparent that MAO-B is capable of oxidizing inert non-polar amines such as MPTP (N-methyl-4-phenyl-1,2,3,6, tetrahydropyridine) and milacemide (2-n pentylaminoacetamide) into neuroactive substances giving rise to Parkinson inducing dopaminergic neurotoxin, MPP+ and inhibitory amino acid neurotransmitter, glycine respectively. These findings accord new prospectives for neuropsychotherapy with selective MAO-B inhibitors and substrates. PMID- 2568665 TI - [New type of homeotic gene with POU domain]. PMID- 2568666 TI - [Rare evolution in a case of Takayasu disease]. PMID- 2568667 TI - [Response of plasma and retinal somatostatin to insulin-induced hypoglycemia in diabetic and control rats]. AB - Changes in plasmatic levels and retinal content of somatostatin after insulin induced hypoglycemia were investigated in three different groups of animals: Control group (C), Diabetic untreated group (D); and, Insulin-treated diabetic group (DI). In addition, another group of animals, not submitted to hypoglycemia, was used as control reference of retinal prehypoglycemic content of somatostatin (group B). Plasmatic basal levels of somatostatin were slightly higher in group DI, and significantly higher in group C, whereas they did not show any differences in group D and DI after hypoglycemia, being significantly higher in group C. The somatostatin retinal content is similar in animals not subjected to hypoglycemia and in the C and DI groups after hypoglycemia, where the rats of the D groups showed significantly higher values than the remainder of the experimental groups, an effect that is also evident in nontreated diabetic animals, even if they are not subjected to hypoglycemia, Summing up, the plasmatic somatostatin response to insulin-induced hypoglycemia is impaired in diabetic rats. Retinal somatostatin content is unchanged after hypoglycemia. PMID- 2568668 TI - Glutathione and its related enzymes in the small intestinal mucosa of rats: effects of starvation and diet. AB - Starvation for 24 h causes a striking fall in glutathione content from 3.19 +/- 0.27 to 1.88 +/- 0.14 (X +/- SEM) mumol/g tissue and of GGT activity from 31.75 +/- 4.17 to 19.49 +/- 3.13 (X +/- SEM) nmol/min/mg protein in the homogenate from whole mucosa of the upper small intestinal segments. This was associated with a significant increase in GSH-Px activity and the content of lipid peroxides (measured by the thiobarbituric assay). On semi-synthetic iron-supplemented diet the activities of GSH-T and GGT were significantly decreased as compared with crude diet. On semisynthetic iron-depleted diet GSH-T and GGT activities were further depressed, but this was accompanied with an additional depression of GSH, glutathione reductase (GSSG-R), and glutathione peroxidase (GSH-Px) activities and lipid peroxide concentrations. Food deprivation significantly lowers the mucosal GSH-content and could lead to a destabilization of this system presumably by increased oxidative stress. As compared to normal "crude" diet, semisynthetic diets and oral iron depletion have been shown to cause a depression of the intestinal GSH system. As a consequence of these effects, the resistance of the small intestinal mucosa toward exogeneous dietary toxins might be reduced. PMID- 2568669 TI - [Acute and chronic effects of bunazosin in patients with congestive heart failure]. AB - The acute hemodynamic effect (right atrial pressure, mean pulmonary artery pressure, pulmonary capillary wedge pressure, cardiac index, heart rate, blood pressure) and neurohumoral response (alpha-ANP, plasma renin activity, aldosterone, angiotensin II) of Bunazosin, oral alpha 1 blocker, was investigated in 28 patients with congestive heart failure at rest and immediately after exercise. Bunazosin reduced alpha-ANP, but, other neurohumoral factors did not change. Bunazosin produced significant hemodynamic improvements both at rest and after exercise. Its chronic effect was also investigated in 11 patients in 28 days after taking oral Bunazosin. Improvement of hemodynamics at acute phase was also preserved at chronic phase without deterioration of neurohumoral factors. It is concluded that Bunazosin may be an effective Balanced vasodilator both at acute and chronic phases in patients with congestive heart failure. PMID- 2568670 TI - Human immunodeficiency virus--associated lung infection: an overview. PMID- 2568671 TI - Stress and the heart. PMID- 2568672 TI - [Various encephalopathies caused by drugs]. AB - Practically all drugs administered in large amounts can give rise to neurologic symptoms such as drowsiness, insomnia, confusion, seizures or coma and extrapyramidal disorders. In this study, five classes of agents are reviewed: antipsychotic drugs, drugs for Parkinson's disease, antiepileptic drugs, calcium antagonists and salts of bismuth. PMID- 2568673 TI - [Muscular diseases in relation to drug consumption]. AB - Toxic myopathies are an uncommon manifestation of chemotherapeutic agents. Most myopathies are characterized by a weakness affecting proximal limb-muscles more than distal ones. Steroids, chloroquine and ipecac syrup are the most common drugs inducing myopathies. Toxic myositis is rare and reported with D penicillamine abuse. Myotoxicity of local agents injection such as anesthetics, steroids or antibiotics is also known. Congenital myopathies such as malignant hyperthermia, hypokalemic periodic paralysis or paroxysmal myoglobinuria may also be induced by drugs but are very rare. PMID- 2568674 TI - [Gastroduodenal ulcers resistant to medical treatment]. PMID- 2568675 TI - The minor problem of hemostatic impairment in nephropathia epidemica, the mild Scandinavian form of hemorrhagic fever with renal syndrome. AB - Nephropathia epidemica (NE) is the mild Scandinavian form of hemorrhagic fever with renal syndrome (HFRS). Its hemorrhagic manifestations are so mild that initially--as its name implies--it was not considered a hemorrhagic fever. It is, however, clinically, histologically, and epidemiologically similar to Korean hemorrhagic fever (KHF) except that hemorrhagic manifestations are fewer and mortality is lower. The etiologic agents of the two disorders, Puumala virus (NE) and Hantaan virus (KHF), are closely related. NE is the most benign of the hemorrhagic fevers. Hemostatic impairment is not a significant clinical problem in this disease and therefore has not been thoroughly studied, although abnormalities in hemostasis have been noted. Further investigations of this disease might provide insights concerning the treatment of other, more severe, hemorrhagic fevers. PMID- 2568676 TI - Hemorrhagic fever with renal syndrome in Korea. AB - Several clinical variants of hemorrhagic fever with renal syndrome (HFRS) are caused by Hantaan and related viruses. Since 1951, 500-900 patients with HFRS have been hospitalized annually in Korea. Although HFRS is associated primarily with rural areas, it is now being recognized as an urban problem and a particular hazard to laboratory staff using rodents for research. Recently, epidemic outbreaks of leptospirosis and scrub typhus have occurred during the HFRS season, leading to confusion in diagnosis. Serologic diagnosis of HFRS is based on the demonstration of IgM antibodies to Hantaviruses by the indirect fluorescent antibody technique or enzyme-linked immunosorbent assay. The specific Hantavirus causing infection can be identified on the basis of titers of plaque-reduction neutralizing antibody. Results of studies with monoclonal antibodies suggest that viral subtypes exist for each Hantaviral serotype presently recognized. While infection with Hantaviruses is known to be a problem of worldwide dimensions, present evidence indicates that it occurs over a wider area than previously recognized. Vertical transmission of Hantaan virus in a pregnant woman has been documented. PMID- 2568678 TI - Hemorrhage in hemorrhagic fever with renal syndrome in China. AB - Hemorrhage is a prominent feature of hemorrhagic fever with renal syndrome (HFRS) in China. It occurs in all phases of the disease and is an important cause of death. Petechiae involving skin and oropharyngeal mucosa are the commonest manifestation of hemorrhage, occurring in more than 90% of patients. Gastrointestinal hemorrhage is the next commonest manifestation, occurring in approximately 50% of patients. Suggested mechanisms of hemorrhage include vascular injury, thrombocytopenia and platelet dysfunction, disseminated intravascular coagulation, circulating heparin-like activity, and uremia. Controlled trials of treatment regimens for hemostatic impairment in HFRS have not been performed. Support of blood pressure can lessen hemorrhage by limiting the adverse consequences of hypotension and shock. Dialysis is of benefit in patients with hemorrhage and significant renal failure. PMID- 2568677 TI - Coagulopathy in hemorrhagic fever with renal syndrome (Korean hemorrhagic fever). AB - The pathophysiology of bleeding manifestations in hemorrhagic fever with renal syndrome (HFRS) was elucidated by serially evaluating coagulation and fibrinolytic profiles and complement alterations in patients with HFRS. In the early stage of the disease, platelet counts, platelet survival time, and platelet aggregation in vitro decreased. Prolongation of bleeding time, prothrombin time, and activated partial thromboplastin time was noted, with decreases in coagulation factors II, V, VIII, IX, and X. Levels of fibrinogen were decreased, and those of fibrinogen-fibrin degradation products in serum and urine were increased. Concentrations of plasminogen, alpha 2-plasmin inhibitor, and antithrombin III in plasma were depressed. Procoagulant activity was present in plasma. Circulating immune complexes were found, whereas serum levels of C3 were decreased. In the early stage of HFRS, thrombocytopenia, defects in platelet function, and disseminated intravascular coagulation may play central roles in the pathogenesis of bleeding manifestations. Vasculopathy and immunologic aberrations also may play a role. PMID- 2568679 TI - Hemorrhagic fever with renal syndrome in Greece: clinical and laboratory characteristics. AB - The clinical and laboratory characteristics of a severe form of hemorrhagic fever with renal syndrome (HFRS) in Greece are presented. Twenty-seven patients with serologically confirmed HFRS were studied; 10 required renal dialysis, six had hemorrhagic manifestations, and four died. In patients with hemorrhagic manifestations, the platelet counts were generally less than 100,000 cells/microL. In three patients findings were compatible with disseminated intravascular coagulation. Laboratory investigation showed a consistent rise in levels of serum urea nitrogen and creatinine beginning on the fifth or sixth day of illness and reaching a maximum level between the ninth and 12th days of illness. The disease in Greece more closely resembles the Asian form of HFRS (Korean hemorrhagic fever) than the Scandinavian form of the disease (nephropathia epidemica) because of the high mortality rate, the occurrence of hemorrhagic manifestations, and the severity of the clinical disease. PMID- 2568680 TI - An idiopathic febrile necrotizing arteritis syndrome in the dog: beagle pain syndrome. AB - The clinical, laboratory, and pathologic features of a syndrome in dogs characterized by intermittent pain, fever, neutrophilia, and necrotizing arteritis are described to alert others involved in toxicity testing to the existence of this disorder. It is considered that this idiopathic syndrome is a latent condition, the expression of which can be precipitated in predisposed dogs by experimental treatment, and thus, its occurrence could complicate interpretation of toxicity studies. We have observed the disorder in 14 beagle dogs. The syndrome is rare and most cases for study were supplied by the breeder. Typical clinical signs observed included evidence of pain when the mouth was opened, grunting when lifted, and standing with an arched back and lowered head. Appetite was usually reduced. Body temperature was elevated (e.g., 104-106 degrees F). There was progressive, bilateral atrophy of temporal and cervical musculature. Such signs have been observed to persist unremittingly or, more commonly, with periods of expression and remission. Neutrophilic leukocytosis and thrombocytosis were present. Hemoglobin and hematocrit were usually slightly decreased. Serum total protein was usually normal but albumin was reduced and alpha-2 globulins were markedly increased. Rheumatoid factor was elevated in several dogs. Arteritis was observed histologically and was characterized by necrosis, intimal proliferation, neutrophil and mononuclear cell infiltration in the media and periarterial tissues, and hemorrhage. Amyloidosis was observed in several dogs. The cause of this disorder is unknown. Knowledge of the distinct features of this syndrome should obviate complication of interpretation of results in toxicity studies and hopefully will lead to studies of this syndrome to provide an understanding of its etiopathogenesis. PMID- 2568681 TI - Pathogenesis of cardiovascular alterations in dogs treated with minoxidil. AB - Minoxidil and other potent vasodilators cause coronary arterial injury, right atrial hemorrhagic lesions, and subendocardial necrosis in dogs. This paper discusses the pathogenesis of coronary arterial and right atrial lesions associated with minoxidil in the dog. Acute coronary vascular injury characterized by segmental medial hemorrhage and necrosis and perivascular inflammation occurred only during the first few days of treatment, after which tolerance to further acute injury developed. At 30 d or more of treatment, coronary vascular injury was characterized by perivascular fibrosis rarely attended by medial distortion or hyperplasia and subintimal thickening, changes consistent with responses to previous injury. Right atrial hemorrhagic lesions, unlike coronary vascular injury, often became progressively more extensive with continued treatment. At 3 d, atrial hemorrhage and inflammation were confined to the subepicardium of the right atrium, evidently around affected subepicardial branches of the right coronary artery. At 30 d, fibrovascular proliferative right atrial lesions (granulation tissue with evidence of continual hemorrhage) extended from the epicardium to the myocardium, with eventual replacement of the atrial wall by mature connective tissue at 1 yr of treatment. Minoxidil-induced cardiovascular lesions were not prevented by treatment with a beta-blocker (propranolol), or an alpha-blocker (dibenzylene), or by sympathetic neural activity suppression (surgical sympathectomy or constant carotid sinus nerve stimulation), suggesting that the sympathetic response to the pharmacologic activity of minoxidil was not responsible for the induction of the cardiovascular lesions. Minoxidil-related vascular lesions were confined to the most pharmacologically responsive segment of the arterial system, the coronary arteries, suggesting that medial injury may have been associated with tensile changes in the arterial wall. PMID- 2568683 TI - [Current status of rheumatology in the North-West. "ARNO 88". Current status of pathology of the cartilage. Caen, 28 October 1988]. PMID- 2568682 TI - Pathogenesis of arterial lesions induced by dopaminergic compounds in the rat. AB - Fenoldopam mesylate (FM), a selective post-junctional dopaminergic (DA1) vasodilator, causes lesions of large caliber splanchnic arteries (100-800 microns) in the rat characterized by necrosis of medial smooth muscle cells and hemorrhage. FM does not induce lesions in other vascular beds of the rat, or in dogs or monkeys. Dopamine, like FM, causes hemorrhagic lesions of large caliber splanchnic arteries in the rat, as well as fibrinoid necrosis of small caliber arteries (less than 100 microns) of the splanchnic, cerebral, coronary and renal vascular beds. Dopamine is an alpha- and beta-adrenoceptor and a dopaminergic receptor agonist. Because these arterial lesions are thought to result from the pharmacologic activity of these 2 compounds, we sought to ascertain the presence of DA1 receptors in mesenteric arteries of the rat and to determine the role of these or other vascular receptor subtypes in lesion induction. We also studied the process of repair after arterial injury caused by FM or dopamine. The presence of DA1 receptors was confirmed in isolated perfused mesenteric arteries by standard pharmacologic techniques; stimulation by FM resulted in vasodilation which was inhibited by the DA1 receptor antagonist SK&F 83566-C. Likewise, SK&F 83566-C prevented the induction of hemorrhagic lesions of large caliber arteries in rats upon infusion of FM or dopamine. In rats co-exposed to the alpha adrenoreceptor antagonist phenoxybenzamine (PBZ) and either FM or dopamine, the incidence and severity of hemorrhagic lesions of large caliber arteries were increased, but PBZ prevented the formation of dopamine-induced fibrinoid lesions in arteries of small caliber. Rats exposed concurrently to dopamine, phenoxybenzamine, and SK&F 83566-C were free of all arterial lesions. Thus, the induction of splanchnic arterial lesions in the rat by dopamine and FM is caused by stimulation of, and interaction between, alpha-adrenoceptors and dopaminergic DA1 receptors. Fibrinoid lesions of small arteries (alpha-adrenoceptor-mediated) were repaired, as observed morphologically by 14 d after exposure to dopamine. Hemorrhagic lesions of large caliber arteries (DA1 receptor-mediated) had undergone significant repair by 28 d after exposure to FM but these arteries possessed a thicker media surrounded by adventitial fibrosis. Thus, morphologically distinct receptor-mediated splanchnic arterial lesions induced by dopaminergic and alpha-adrenoceptor agonists follow a markedly different course of repair. Arterial lesions induced by FM or dopamine by activation of post junctional dopaminergic DA1 receptors may represent a model of polyarteritis nodosa. PMID- 2568684 TI - Colloidal bismuth in the treatment of duodenal ulceration: the benefit for the patient. AB - Colloidal bismuth subcitrate (CBS) heals gastric and duodenal ulcers better than placebo and at rates similar to those observed with the commonly used H2 antagonists. Indeed, healing of duodenal ulcers may be more rapid than with cimetidine during the first month of treatment. When treatment is withdrawn, however, relapse is slower after CBS than after the H2-antagonists. The mechanism is uncertain but may be related to the ability of CBS to suppress Campylobacter pylori infection. The clinical implications of this difference in relapse rates are discussed. Preliminary data suggest that duodenal ulcer patients who are rendered C. pylori-negative for a prolonged period may be relatively immune to relapse. If confirmed, and if a suitably effective regimen can be found, this will transform the management of this difficult clinical problem. PMID- 2568686 TI - The gastrointestinal tract in growth and reproduction. PMID- 2568685 TI - [A new beta 1-receptor blocker in the therapy of essential hypertension and angina pectoris]. AB - In 242 patients with hypertension and/or angina pectoris, a new cardioselective betablocker without ISA, bisoprolol (Concor), was tested. The average mean value of 168/102 mm Hg was lowered in the 174 hypertensive patients by a systolic value of 17 and a diastolic value of 11 mm Hg. A normal diastolic pressure of 95 mm Hg or below was attained within 4 weeks in 73% of patients. Angina pectoris improved from 7 attacks per week before treatment to 3 attacks after 2 weeks; patients with additional hypertension showed a further improvement after another two weeks to an average of 1.7 attacks per week. Side effects were most frequently dizziness, headache and fatigue and also a few patients with gastrointestinal symptoms, an unusual side effect with this treatment. The results show the effective antihypertensive and antianginal action of bisoprolol in a large group of outpatients. PMID- 2568687 TI - [A soiled woman...]. PMID- 2568688 TI - [Pharmacology and mechanism of action of anxiolytic agents]. PMID- 2568689 TI - [Treatment of anxiety]. PMID- 2568690 TI - [Anxiety and depression]. PMID- 2568691 TI - [An aversion for leather...]. PMID- 2568692 TI - [The asylum--creation, expansion and death?]. PMID- 2568694 TI - Activity of gamma-GT and content of glutathione in xenotransplanted squamous cell carcinomas under cisplatin chemotherapy. PMID- 2568693 TI - Craniotomy improves outcome in subdural empyema. AB - Renewed uncertainty regarding the best technique of surgical therapy (burr holes versus craniotomy) for subdural empyema prompted a review of the experience with this disease entity at UCLA. Based on data obtained from this review and from studies previously published in the literature, the keys to optimal outcome are rapid diagnosis, craniotomy with total removal of pus, and appropriate antibiotic therapy. The clinical syndrome of subdural empyema can overlap that of other intracranial infections; infants and young children present with a different syndrome than older children and adults. Computed tomography scanning is nearly always diagnostic but may be equivocal or normal; magnetic resonance imaging may become the diagnostic study of choice. Multiple organisms are often cultured, which requires multiple drug therapy. PMID- 2568695 TI - Early visual field changes with beta-blocking agents. PMID- 2568697 TI - Cardiovascular and pulmonary effects of beta-blocking agents: implications for their use in ophthalmology. PMID- 2568696 TI - Pharmacologic effects of beta-blocking agents used in the management of glaucoma. PMID- 2568698 TI - Review of clinical studies of beta-blocking agents. PMID- 2568700 TI - [Beta blockaders do not influence the development of arteriosclerosis]. PMID- 2568699 TI - Pressure compliance test of the optic nerve head: influence of different antiglaucoma drugs. PMID- 2568701 TI - Medetomidine and atipamezole in small animal practice. PMID- 2568702 TI - Effects of methylmercury on neurotransmitter release from rat brain synaptosomes. AB - Although the effects of methylmercury (MeHg) at the neuromuscular junction have been well characterized, similar studies employing CNS preparations and transmitters have been limited. We found that MeHg (0.5-5.0 microM) produced a concentration-dependent increase in the spontaneous release of [3H]dopamine. gamma-[3H]aminobutyric acid, and [3H]acetylcholine from synaptosomes isolated from rat brain striatum, cortex, and hippocampus, respectively. At these same concentrations MeHg did not attenuate calcium-dependent depolarization-evoked 3H transmitter release. MeHg did not appear to induce calcium influx into the nerve terminal since the increase in release persists in the absence of extrasynaptosomal calcium. The increase in spontaneous transmitter release induced by MeHg persisted in the presence of low extrasynaptosomal sodium, suggesting that MeHg's effects on release are not mediated by either Na+, K+ ATPase inhibition or selective increases in membrane sodium permeability. MeHg produced only a very small increase in 45Ca efflux from synaptosomes preloaded with 45Ca, whereas these same MeHg concentrations produced large increases in 45Ca efflux from preloaded isolated mitochondria. MeHg did increase the efflux of [3H]deoxyglucose phosphate from synaptosomes. An increase in the efflux of [3H]deoxyglucose phosphate is believed to reflect an increase in neuronal membrane permeability. The quantitative and temporal aspects of the MeHg-induced [3H]-deoxyglucose phosphate efflux were similar to those observed for MeHg induced neurotransmitter release. These data suggest that the increase in spontaneous transmitter release induced by MeHg is mainly the result of transmitter leakage that occurs subsequent to MeHg-induced increases in synaptosomal membrane permeability. However, these results cannot exclude possible effects of MeHg on intrasynaptosomal calcium homeostasis. PMID- 2568703 TI - Comparative study of amino acid composition in an extract from hornet venom sacs: high content of neuroactive amino acids in Vespa. AB - The amino acid compositions of extracts from the venom sacs of Vespa mandarinia, V. xanthoptera, V. tropica, V. analis and Vespula lewisi were analyzed. omega Amino acids, which are inhibitory neurotransmitters, such as gamma-amino-n butyric acid, taurine, beta-alanine and glycine, were predominant components in Vespa, but a minor component in Vespula. Glutamic acid, an aminergic excitatory neurotransmitter, was present in large quantities. Leucine, an insect autotoxin, was detected. Arginine was found in all the venoms and glutamine was also abundant. Tryptophan and histidine, precursors of serotonin and histamine, respectively, were also found. Thus, neuroactive amino acids that may exert inhibitory effects on insects neuronal axons and neuromuscular junctions were present in large amounts, and may facilitate paralysis of insect prey. PMID- 2568704 TI - Serum succinyltrialanine p-nitroanilide-hydrolytic activity in workers occupationally exposed to lead. AB - The relationship between blood lead concentration and serum succinyltrialanine p nitroanilide-hydrolytic (STN) activity was investigated in 74 workers occupationally exposed to lead and in 28 non-exposed workers. Exposure to lead was observed during transfer-printing processes in which paints or paint powders containing 20-50% lead were used. The mean lead concentration was 0.49 mg/m3 (0.12-1.43 mg/m3) in the working environment and 1.42 mg/m3 (0.21-4.34 mg/m3) in workers involved in printing processes. Serum STN activity became lower with increasing concentrations of blood lead (PbB) in non-exposed workers (controls). In lead-exposed workers, the activity decreased with increasing concentrations of PbB of 2.0 mumol/l and more, but increased when less than 1.9 mumol/l. Among lead exposed workers with high PbB concentrations and/or with hepatic dysfunction, the STN/PbB ratio was distributed above the regression line obtained from the controls. The present investigation suggests that serum STN activity decreases in those highly exposed to lead. Enzymes such as elastase, which hydrolyzes succinyltrialanine p-nitroanilide and additionally is not inhibited by lead, may possibly be induced among lead workers when liver function is impaired. PMID- 2568705 TI - Effect of mild hypothermia on ischemia-induced release of neurotransmitters and free fatty acids in rat brain. AB - We have demonstrated previously that mild intraischemic hypothermia confers a marked protective effect on the final histopathological outcome. The present study was carried out to evaluate whether this protective effect involves changes in the degree of local cerebral blood flow reductions, tissue accumulation of free fatty acids, or alterations in the extracellular release of glutamate and dopamine. Rats whose intraischemic brain temperature was maintained at 36 degrees C, 33 degrees C, or 30 degrees C were subjected to 20 minutes of ischemia by four vessel occlusion combined with systemic hypotension. Levels of local cerebral blood flow, as measured autoradiographically, were reduced uniformly in all experimental animals at the end of ischemia by gas chromatography after tissue extraction and separation by thin layer chromatography. A massive ischemia induced accumulation of individual free fatty acids was observed in animal groups whose intraischemic brain temperature was maintained at either 36 degrees C or 30 degrees C. Extracellular neurotransmitter levels were measured by microdialysis; the perfusate was collected before, during, and after ischemia. In rats whose intraischemic brain temperature was maintained at 36 degrees C, dopamine and glutamate increased significantly during ischemia and the early period of recirculation (by 500-fold and sevenfold, respectively). In animals whose brain temperature was maintained at 33 degrees C and 30 degrees C, the release of glutamate was completely inhibited, and the release of dopamine was significantly attenuated (by 60%). These results suggest that mild intraischemic hypothermia does not affect the ischemia-induced local cerebral blood flow reduction or free fatty acid accumulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568706 TI - Characteristics of a DNA probe (pa3'HVR) when used for paternity testing. AB - DNA probes that detect polymorphic loci in the human genome are finding widespread application in many areas of genetic testing. Paternity testing represents one area for the application of probe technology; this report presents data obtained in a paternity testing program with a probe (pa3'HVR) derived from a locus (D16S85) approximately 8 kilobases (kb) downstream from the alpha globin gene complex on chromosome 16. The pa3'HVR probe used under stringent conditions of hybridization detects a highly polymorphic locus in chromosomal DNA digested with Pvu 2 restriction endonuclease. Alleles at the D16S85 locus were grouped into 58 size bins differing from one another by 100 base pairs in the black and white populations. The most common alleles detected in whites fell into the 2.3 kb group with a collective frequency of 0.1849. In blacks, the most common allele group is 2.0 kb with a collective frequency of 0.1333. The probe was used for restriction fragment length polymorphism mapping in conjunction with standard paternity testing techniques in 100 paternity cases. Thirty direct exclusions were encountered in the 100 cases with standard testing methods, versus 27 exclusions with the pa3'HVR probe alone. Four exclusions detected with standard methods were not detected with the probe and one exclusion detected with the probe was missed by standard testing. The probability of excluding a falsely accused man by use of the pa3'HVR probe was approximately 90 percent. In cases where exclusions were not encountered, the data obtained with the pa3'HVR probe increased the paternity index calculated from standard testing by about 16 fold.2+ informative for paternity testing. PMID- 2568707 TI - Hematopoietic reconstitution using progenitors recovered from blood. PMID- 2568708 TI - Use of cDNA probes specific for the human MHC class II beta loci for tissue typing nonhuman primates at their class II beta loci. PMID- 2568709 TI - Presynaptic nicotinic autoreceptors. PMID- 2568710 TI - [Regional, cellular, subcellular distribution of neurospecific protein-hormonal complexes]. AB - A sensitive radioimmunological assay (RIA) has been developed to detect the tissue specificity and subcellular localization of three specific protein hormonal complexes (PHC) of the hypothalamus which have a regulatory function in the brain and visceral organs. Using the highly specific rabbit antisera to beef PHC an order of increasing immunoreactivity in different areas of CNS is as follows: hypothalamus, cerebellum, occipital cortex. The PHC-like immunoreactivity (IR) is found in the neurosecretory granules of the hypothalamo neurohypophyseal system (about 30%) and in the synaptosomal fraction (70%). In the myelin, mitochondrial and nuclear fractions the PHC-like IR is not revealed. IR of PHC is 1000-fold higher in the brain than in visceral organs (heart, skeletal muscles, adrenals, pancreas and blood serum). A possible role of the PHC as markers of the neuroendocrine cells is discussed. PMID- 2568711 TI - Does medical therapy cure female stress incontinence? AB - Forty-four consecutive female patients, who were scheduled for operation for genuine stress incontinence but instead treated pharmacologically with an alpha agonist or placebo for 3-4.5 months, were evaluated after a median observation period of 30 months. Ten patients (23%) underwent colposuspension. Fourteen patients (32%) claimed to be continent or so much improved that further treatment was considered unnecessary by the patients themselves. Of the remaining 20 patients who still claimed to have stress incontinence, 11 (25%) preferred to resume the pharmacological treatment whereas 9 patients (20%) had not decided whether they wanted further treatment. These results suggest that some operations for stress incontinence may be avoided by introduction of a standard trial of medical therapy prior to intended surgical intervention. PMID- 2568712 TI - Norfenefrine in the treatment of female urinary stress incontinence assessed by one-hour pad weighing test. AB - Thirty-three women with urinary stress incontinence were treated with the alpha adrenoceptor-stimulating agent, norfenefrine (Nevadral Retard). Treatment was administered during 3-24 weeks at a daily dosage of 90 mg. Judged by a 1-hour pad weighing test, a significant effect could be demonstrated. With the exception of 1 case of allergic reaction and 1 case of recurrent vomiting, a simultaneously performed tolerability test revealed only minor side effects. PMID- 2568714 TI - Beta-agonists as repartitioning agents: a review. AB - Several beta-adrenergic agonist compounds are undergoing commercial development as carcase repartitioning agents. Significant increases in feed conversion efficiency, carcase lean content and decreases in carcase fatness have been reported consistently in all the major meat animal species. The use of these compounds is consistent with consumer demand for leaner meat but could prove controversial in the current political climate in the EC. PMID- 2568713 TI - Preoperative and surgical management of pheochromocytoma. AB - The clinical awareness of the various presentations of pheochromocytoma, along with newer monitoring and anesthetic techniques, has enabled early diagnosis and less fluctuation of blood pressure during operation. The use of meticulous surgical technique through the various incisions outlined in this article has made the excision of pheochromocytoma an expeditious operation and the accompanying hypertension surgically curable. Thus, pheochromocytoma is less likely to be a fatal lesion. PMID- 2568715 TI - Multiple drug-resistant strains of Salmonella typhimurium in poultry. PMID- 2568716 TI - [Clinico-genetic aspects of medullary thyroid cancer]. AB - A complex (population, clinico-familial, cytogenetic and biochemical) investigation was performed in 50 cases of thyroid medullary cancer and their relatives. The study evaluated the diagnostic value of tests used to identify hereditary forms of the disease which accounted for one-third of the cases. Guidelines for identification of groups at high risk for thyroid medullary carcinoma are discussed. PMID- 2568717 TI - Do autoantibodies lead to immune damage in AIDS? PMID- 2568718 TI - [Neurochemical mechanisms of autostimulation]. PMID- 2568719 TI - [Mechanisms of occurrence of dissociated cerebral states]. PMID- 2568720 TI - [Multiple endocrine neoplasia of the II B type]. AB - Multiple endocrine neoplasia, type II B (MEN II B) includes thyroid carcinoma, pheochromocytoma, ganglioneuromatosis and marfanoid habitus. Its short-term prognosis is determined by the tumour of the adrenal gland, whereas long-term prognosis is determined by the thyroid carcinoma. Often the well-defined but rare syndrome is diagnosed late. The MEN II B's therapy consists of thyroidectomy and adrenalectomy uni- or bilateral. The 5 year-rate of survival is 75%. Own observations are reported. PMID- 2568721 TI - [Functional properties of isolated kidney mitochondria of rats in food deprivation]. AB - The respiratory properties of isolated rat renal mitochondria after fasting conditions (2-8 days) are estimated in comparison with the parameters of normal feeded control animals. After an extrem starvation time of 8 days the active and uncoupled respiration of glutamate/malate, but not of succinate were reduced, whereas the respiratory control index and the ADP/O-Quotient did remain unchanged. In conclusion, malnutrition do not influence the renal function due to mitochondrial changes. PMID- 2568722 TI - [AST, GLDH, gamma-GT, total bilirubin and CK values during the first week of life in healthy premature calves or calves with a late asphyxia syndrome]. AB - The physical condition of 44 calves delivered by caesarean section before term was monitored by clinical and repeated laboratory examinations (analysis of AST, GLDH, gamma-GT, CPK, total bilirubin) during the first seven days of life. The newborns were divided into two groups based on the clinical observations during the first hour of life, the blood pH and the base deficit: Group 1: 30 calves without respiratory distress syndrome (vital, non asphyxial); they did not develop any diseases in the course of the experiment. Group 2: 14 calves with respiratory distress syndrome (asphyxial; 9 of these animals died in the course of the experiment. No significant differences between the vital and asphyxial calves were found in respect to the enzymes AST, GLDH, gamma-GT, CK as well as total bilirubin values measured during the first week of life. These blood parameters were within the normal range for calves delivered at term. The results do not indicate any disorder in liver and muscle functions in prematurely born calves with or without respiratory distress syndrome. PMID- 2568723 TI - [Neuromediators of centrifugal innervation of the main olfactory bulb]. PMID- 2568724 TI - Cholinergic stimulation of insulin release from cloned B-cell lines HIT-T15 and RINm5F. AB - Acetylcholine stimulated the release of insulin from cultured HIT-T15 and RINm5F cells in the absence of glucose confirming that the insulin response to cholinergic stimulation is preserved in these two cloned B-cell lines. In addition acetylcholine potentiated glucose stimulated insulin release from HIT T15 but not from RINm5F cells. Perifused HIT-T15 cells responded to square wave acetylcholine challenge with a monophasic release of insulin while the release profile for RINm5F cells was irregular and reduced. Acetylcholine stimulated insulin release from both cell lines was reduced by atropine but unaffected by hexamethonium confirming a mechanism of action involving muscarinic receptor activation. PMID- 2568725 TI - Effects of cyclosporin A on the endocrine pancreas of the rat. Morphological and immunohistochemical studies. AB - Treatment with the immunosuppressive agent cyclosporin A frequently gives rise to functional alteration of the islets of Langerhans which may result in diabetes. Light microscopy, immunocytochemical, and electron microscopy investigations demonstrate degranulation, vacuolization, and destruction of endocrine pancreatic cells in treated animals. Similar changes, but of a milder degree, are observed in the A-cells. The morphological alterations described are likely to be the result of inhibition of synthesis and secretion in B-cells. PMID- 2568726 TI - GHRH treatment: studies in an animal model. AB - This study examined the effects of chronic deletion of circulating growth hormone releasing (GHRH) and/or somatostatin (SRIF) on normal growing male rats, as well as the effects of exogenous GHRH (1-29)NH2 and/or SMS 201-995 administration on the growth of rats with hypothalamic ablation. Passive immunization with anti-rat GHRH goat gamma-globulin (GHRH-Ab) for 3 weeks caused a marked decrease in the levels of pituitary GH mRNA and severe growth failure. Treatment with anti-SRIF goat gamma-globulin (SRIF-Ab) for 3 weeks produced a more modest decrease in GH mRNA levels in the pituitary and a slight but significant inhibition of normal somatic growth. Hypothalamic ablation produced a marked decrease in the level of mRNA in the pituitary. Chronic continuous administration of GHRH (1-29)NH2 stimulated pituitary GH synthesis, elevated serum levels of insulin-like growth factor I and increased body weight gain in rats with hypothalamic ablation treated with replacement doses of cortisone, testosterone and L-thyroxine. Combined treatment with GHRH (1-29)NH2 and SMS 201-995 appeared to promote the effect of GHRH on pituitary GH release and somatic growth in these animals. The results suggest that continuous administration of GHRH will be useful in the treatment of children with growth retardation resulting from hypothalamic disorders. In children with combined GHRH and somatostatin deficiencies, the addition of somatostatin to a GHRH treatment regimen may produce better results. PMID- 2568727 TI - Neuroendocrine control of growth hormone secretion. PMID- 2568728 TI - Adrenergic and non-adrenergic mechanisms in sympathetic vascular control of the nasal mucosa. AB - 1. Co-existence of NA and NPY-LI was observed in the dense network of sympathetic nerves around both resistance and capacitance vessels in the nasal mucosa from most species including man. NPY-LI seems also to be present together with VIP-LI and PHI-LI in periglandular and some perivascular parasympathetic fibres in some species. 2. A new in vivo model was developed in the pig which allowed parallel recordings of arterial blood flow (BF), capacitance function (V) and superficial movement of blood cells (LDF signal) of the nasal mucosa upon SNS and/or local intra-arterial injection of various pharmacological agents. SNS reduced both BF and V. The V response was maximal already at low frequency. The LDF signal was increased when the SNS-evoked BF reduction was under 40% but was reduced upon larger BF response. After pretreatment with the alpha-adrenoceptor antagonist PBZ, the vascular responses to SNS were significantly attenuated but clear-cut reductions of BF, V and LDF responses remained even to a single impulse. Exogenous NA induced dose-dependent reductions of the BF, V and LDF signal, which were abolished by PBZ treatment. ATP caused biphasic vascular effects with short lasting vasoconstriction followed by vasodilatation. NPY and mATP caused reduction of the vascular parameters via non-adrenergic mechanisms. 3. A frequency-dependent increase of NA overflow was observed in the nasal venous effluent upon SNS in control pigs, whereas detectable release of NPY-LI occurred only at high frequency. Pretreatment with PBZ significantly increased the SNS evoked NA and NPY-LI overflow, while DMI enhanced both the functional response and NA release but reduced the NPY-LI overflow. These data suggest that NA and NPY release in the pig nasal mucosa is regulated by prejunctional alpha adrenoceptor mechanisms. During tachyphylaxis to mATP, the NA and NPY-LI overflow and the vasoconstrictor responses to SNS were unmodified in controls. 4. Reserpine pretreatment induced a marked depletion of both NA and NPY-LI in the pig nasal mucosa. The reduction of NA was not influenced by preganglionic denervation while the depletion of NPY then was prevented. Slowly developing, frequency-dependent and long-lasting vascular responses remained (up to 80% of control) upon SNS in reserpinized and decentralized animals. In these animals, the NA overflow was abolished while release of NPY-LI was enhanced. Stimulation with irregular bursts, caused both larger vascular responses and more NPY-LI overflow in comparison with continuous stimulation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2568729 TI - Alpha 2-adrenoceptor agonist-mediated inhibition of [3H]noradrenaline release from rat hippocampus is reduced by 4-aminopyridine, but that caused by an adenosine analogue or omega-conotoxin is not. AB - The inhibitory effect of an adenosine analogue, R-PIA, and an alpha 2 adrenoceptor agonist, UK 14,304, on [3H]NA efflux from field-stimulated rat hippocampal slices was examined. The effect of 0.1 microM UK 14,304 was mimicked by 30 nM omega-conotoxin and by 10 microM cadmium chloride, inhibitors of N- and L-type Ca2+ channels. R-PIA (1 microM) had no effect per se, but caused a clear cut inhibition after blockade of the pre-synaptic alpha 2-receptor by yohimbine. 4-Aminopyridine (4-AP) caused a dose-dependent increase in evoked transmitter release. At 30 microM 4-AP did not affect the actions of omega-conotoxin or cadmium chloride. The pre-synaptic effect of R-PIA was similarly unaffected by 30 microM 4-AP. The pre-synaptic effect of UK 14,304 was virtually abolished by 4-AP (30 microM). The effect of UK 14,304 (0.1 microM) could be partly restored by reducing the Ca2+ concentration during treatment with 4-AP (22% inhibition compared to 42% with normal Ca2+). The magnitude of increase in evoked [3H]NA efflux by yohimbine (1 microM) was decreased by 4-AP in a concentration-dependent manner from 142% increase in controls to 21% at 100 microM 4-AP. The present results indicate that NA release is reduced by somewhat different mechanisms by pre-synaptic alpha 2- and adenosine A1-receptors. Furthermore, the results indicate that pre-synaptic A1-receptors on hippocampal NA neurons do not primarily regulate 4-AP-dependent potassium channels, but they might act directly on a Ca2+ conductance. PMID- 2568730 TI - ATP as a sympathetic co-transmitter in rat vasomotor nerves--further evidence that individual release sites respond to nerve impulses by intermittent release of single quanta. AB - A combination of intra- and extracellular recording was used to study neurotransmitter release in sympathetic vasomotor fibres in rat femoral and mesenteric arteries. The results show that neurotransmission in these preparations is similar to that in the 'short' sympathetic nerves in guinea-pig vas deferens and the 'long' sympathetic nerves in rat tail artery in the following respects: (1) The intracellularly recorded excitatory junction potentials (EJPs) and the extracellularly recorded junction currents (EJCs) presumably are caused by ATP secreted as a sympathetic co-transmitter. (2) The stimulus-evoked and spontaneous EJCs share the same brief time-course, which is similar to that of spontaneous EJPs, but much briefer than that of stimulus evoked EJPs. (3) 'Successful' nerve impulses appear to release single transmitter quanta. (4) The probability of activation of individual release sites is low (0.002-0.02). (5) The low release probability cannot be accounted for by failure of the nerve impulse to invade the terminals. Moreover, it is also shown that application of tetrodotoxin to the medium within the recording electrode effectively abolishes transmitter secretion in the area enclosed by the tip of the electrode, indicating that the effective length constant for a passively propagating nerve action potential is probably very small and that activation of the release mechanisms in 'long' sympathetic nerve fibres seems to require that the varicosities are actively invaded. PMID- 2568731 TI - Effects of adrenaline and prior exercise on the release of alanine, glutamine and glutamate from incubated rat skeletal muscle. AB - Catecholamines have been proposed as important regulators of the rate of amino acid release from skeletal muscle. In the present study, we have investigated the influence of adrenergic action and its possible interaction with exercise on muscle release and tissue content of alanine, glutamine and glutamate. For this purpose epitrochlearis muscles were dissected from resting and exercised (1 and 2 h) rats and incubated for 1 h in the presence or absence of adrenaline. In addition, muscles from water-immersed resting rats were included to separate the influence of the stress involved in the swimming exercise from that of muscle contractile activity per se. In muscles from untreated resting rats, the release, tissue content and total amount (released amount + tissue content) of the three amino acids were not influenced by 10(-7) M adrenaline; when the adrenaline concentration was raised to 10(-5) M only the tissue content of glutamate was significantly changed (-50%, P less than 0.001). However, in muscles of rats subjected to 2 h prior exercise or water immersion, 10(-7) M adrenaline significantly increased the release of glutamine (+ 48% and +34%, P less than 0.05) and glutamate (+38% and +27%, P less than 0.05). Moreover, 1 h of water immersion resulted in a significant increase in muscle glutamine and glutamate compared to values from the exercised and control rats. The data suggest that adrenergic action is involved in the regulation of muscle amino acid transport during exercise and that the stress involved in exercise may mask the influence of contractile activity per se on formation of amino acids in skeletal muscle. PMID- 2568732 TI - Comparative effects of the alpha-adrenoceptor agonists noradrenaline, phenylephrine and clonidine in the human saphenous vein in vivo and in vitro. AB - The alpha-adrenoceptor-mediated contractile effects of noradrenaline (alpha 1 + alpha 2), phenylephrine (alpha 1) and clonidine (alpha 2) on human saphenous veins were investigated in vivo and in vitro. By infusion (0.3 ml min-1) of the drugs (increasing concentrations in the infused solution) into distended (40 mmHg) saphenous veins in six healthy subjects, local vasoconstriction was induced, measured by a photo-electric device. The drugs induced dose-dependent contractions in all subjects, and dose-response curves were constructed. These were compared with concentration-response curves based on in-vitro results. Macroscopically normal saphenous veins, taken at saphenousectomies, were used, and the preparations were contracted isometrically in organ baths by the agonists. From the curves obtained in vivo and in vitro, the relative potencies of phenylephrine and clonidine (in relation to noradrenaline) were calculated and compared. The relative potencies of phenylephrine in vivo (76%) and in vitro (82%) did not differ significantly. However, the relative potency of clonidine was significantly (P less than 0.05) lower in vivo (90%) than in vitro (99%). Thus, it is concluded that there are differences between the results obtained in vivo and in vitro, stressing the importance of comparative in vivo-in vitro studies. PMID- 2568733 TI - The 5-HT2 antagonist ritanserin blocks the effect of pre-frontal cortex inactivation on rat A10 dopamine neurons in vivo. PMID- 2568734 TI - Potential mechanism of pulmonary arterial relaxation and guanylate cyclase activation by 15-hydroperoxyeicosatetraenoic acid. PMID- 2568735 TI - Synthesis and functions of cyclooxygenase and lipoxygenase products in brain: new findings and an appraisal. PMID- 2568736 TI - Excitatory effect of histamine on EEGs of the cortex and thalamus in rats. AB - When low frequency electrical stimulations were repeatedly applied to the rat midbrain reticular formation, the EEG spectral power recorded at the cortex and thalamus increased markedly, especially in the low frequency bands (0-6 Hz). The intraventricular administration of histamine (Hi) inhibited this increase. Hi induced inhibition was antagonized by simultaneous injection of an equimolar dose of either pyrilamine or diphenhydramine, but not by that of cimetidine. In accordance with this, 2-methylHi decreased the power in the slow wave region, while 4-methylHi was not effective. It was assumed that the EEG arousal effect of Hi is exerted via H1 receptors but not related to H2 receptors. Adverse effects of H1 blockers on the brain, such as drowsiness, may be produced as a consequence of this inhibitory action. PMID- 2568737 TI - Differential regulation of membrane receptors sensitive to histamine (H2-type), isoproterenol (beta 2-type) and glucagon-like peptides by the somatostatin analogue Sandostatin in rat gastric glands. AB - The potential use of somatostatin-14 and its long lasting analogue Sandostatin as antiulcer agents led us to study the functional properties of these peptides on the histamine H2-receptor H2R adenylate cyclase system in gastric glands isolated from the rat fundus. The action of the two peptides has also been compared on membrane receptors sensitive to isoproterenol and the truncated glucagon-like peptide TGLP-1. The data indicate that somatostatins inhibit selectively H2R and TGLP-1 receptor activity with similar potencies and kinetics, suggesting that the two peptides share the same receptor pool coupled with the Gi subunits of adenylate cyclase. Somatostatin-14 and Sandostatin have no evident action on the beta 2-type adrenergic receptor beta 2R. Therefore, the higher potency of Sandostatin compared to somatostatin-14 in inhibiting acid secretion is probably related to an increased stability of the analogue in vivo. PMID- 2568739 TI - Influence of urea-equivalent groups in position 5 of 2-amino, 2-(1 aminoethylidenamino) and 2-guanidino thiazole derivatives on H2-receptor antagonist activity in gastric fistula cat. AB - A series of thiazole derivatives, in which a side-chain with different urea equivalent groups was introduced in position 5 of the heterocycle, have been tested as inhibitors of dimaprit-induced gastric acid and pepsin secretion in the gastric fistula cat. By comparing the in vivo and the previously reported in vitro activity of these compounds, we can note a very close parallelism not only in the quality of their action but also in the estimates of pA2 values. These data support an interdependence between the molecular substructures and the affinity for the H2-receptor. PMID- 2568738 TI - The effects of cimetidine, ranitidine and famotidine on rat hepatic microsomal cytochrome P-450 activities. AB - It has been questioned whether the interaction of H2-antagonists with cytochrome P-450 that is observed in vitro is also relevant for the in vivo situation. Until now the possibility that cytochrome P-450 may function with different modes of action has been neglected in this respect. We studied the effect of cimetidine, ranitidine and famotidine on the monoxygenase, the oxidase and the peroxidase action of cytochrome P-450. Biotransformation catalyzed by the monoxygenase and oxidase action of cytochrome P-450 was affected by cimetidine (probably via its ligand interaction with cytochrome P-450), whereas metabolism by the peroxidase mode of action of cytochrome P-450 was hardly influenced. Ranitidine and famotidine (both pharmacodynamically more potent than cimetidine) only slightly affected cytochrome P-450 activities. PMID- 2568740 TI - H2-receptor antagonist activity of N-methylthiourea, N-cyano-N'-methylguanidine, N-cyanoamidine, N-carbamoylamidine and N-sulfamoylamidine 5-substituted thiazole derivatives on guinea-pig atria. AB - The H2-antagonist activity of thiazole derivatives, substituted on position 5 with urea-equivalent groups, has been tested on guinea-pig isolated atria stimulated by dimaprit. By comparing the activities of the 2,5-disubstituted thiazole derivatives with those of the corresponding 2,4-disubstituted derivatives it can be seen that the side-chain position is critical to activity and differently influences activity in the various series. The heteroaromatic ring atom sequence N-C-S-C-side chain is always associated with a low antagonist activity. PMID- 2568741 TI - Food-induced histaminosis under diamine oxidase (DAO) blockade in pigs: further evidence of the key role of elevated plasma histamine levels as demonstrated by successful prophylaxis with antihistamines. AB - Using a recently established porcine model, it was clearly shown that oral histamine administration is extremely dangerous in the presence of diamine oxidase (DAO) blockade. Due to the severity of the symptoms (20% death) and the clinical relevance, further interest has been focussed on strategies to prevent or alleviate food induced histaminosis. In a randomized controlled trial, 10 pigs under DAO blockade were challenged with oral histamine (60 mg). Half of these animals received a prophylactic premedication with a combination of H1- and H2 receptor antagonists. As expected, all animals developed a massive increase in plasma histamine levels, with significantly higher values in the control group (median: 123 ng/ml) compared to the antihistamine group (median: 32 ng/ml). In contrast, clinical symptoms were only observed in the control group. The maximum fall in mean arterial pressure (hypotension) was 60 mmHg (median for control group) but only 15 mmHg (median) under antihistamine pretreatment. These results firstly provide further evidence for the causal role of histamine in the new disease concept and secondly enable us to investigate appropriate therapeutic measures for patients at risk. PMID- 2568742 TI - Effect of histamine on the T-cell colony formation of PHA-stimulated cells. AB - The effect of histamine on T-cell colony formation was studied in human peripheral blood mononuclear cells. Histamine inhibited dose-dependently (10(-4) 10(-6) M) the colony formation of PHA-stimulated T-cells. The inhibition was similar in normal controls and rheumatoid arthritis (RA) patients in spite of the fact that in RA the colony formation was significantly lower than in the normal controls. No increase of colony formation was observed at low concentrations (less than 10(-7) M). Impromidine was less effective than histamine, and pyridylethylamine (PEA) was inactive. Cimetidine counteracted the effect of histamine while chlorpheniramine did not. The results show that colony formation may be inhibited through H2-receptors. This action may be of importance in cellular interactions in tissues with high local histamine concentrations. PMID- 2568743 TI - Effect of beta-adrenoceptor blocking drugs on 32P incorporation into and arachidonic acid liberation from phospholipids in stimulated rat mast cells. AB - The lipophilic beta-adrenoceptor blocking (BAB) drugs metipranolol, propranolol and exaprolol significantly decreased 48/80- and A23187-induced 32P incorporation into rat mast cell phospholipids. Exaprolol was the most active, followed by propranolol and metipranolol. Atenolol and metipranolol significantly decreased the 48/80-stimulated, and metipranolol and exaprolol the A23187-stimulated 3H arachidonic acid liberation from isolated mast cells. PMID- 2568745 TI - Antiarrhythmic efficacy of solitary beta-adrenergic blockade for patients with sustained ventricular tachyarrhythmias. AB - To assess the efficacy and predictability of solitary beta-adrenergic blocker (BB) therapy for ventricular tachyarrhythmia (VT), 30 patients (16 men and 14 women) with a mean age of 55 years, who initially had sustained ventricular tachycardia (70%) or ventricular fibrillation (30%), were studied. Results of baseline arrhythmia tests showed VT on ECG monitoring in 57% of the patients, during exercise in 50%, induced by programmed stimulation in 69%, increasing to 86% during isoproterenol. BB therapy prevented inducible VT during programmed stimulation in 37% of the patients, prevented VT on ECG monitoring in 54%, and prevented VT during exercise in 83%. Long-term BB therapy was given to 24 of 30 patients, whereas six other patients with hemodynamically unstable VT during BB therapy received other long-term treatment. During a mean follow-up of 824 days, 6 of 24 patients had recurrent VT. BB therapy was discontinued in two patients because of side effects. Long-term success was predicted by left ventricular ejection fraction greater than 45%, absence of coronary disease, and age less than 60 years (all p less than 0.02). Neither suppression of arrhythmia during exercise testing, nor results of programmed stimulation or ECG monitoring were predictive of outcome. Thus beta-adrenergic blockers can be effective as solitary antiarrhythmic therapy in selected patients with VT. PMID- 2568744 TI - Comparison of coronary angioplasty with medical treatment for single- and double vessel coronary disease with left anterior descending coronary involvement: long term outcome based on an Emory-CASS registry study. AB - The 3 to 5-year clinical outcomes of 627 consecutive patients selected for treatment with percutaneous transluminal coronary angioplasty (PTCA) at Emory University Hospital for one- or two-vessel coronary artery disease with involvement of the left anterior descending coronary artery, between July 1981 and June 1983, and 865 patients in the Coronary Artery Surgery Study with similar coronary artery disease involvement selected for medical therapy between 1975 and 1979 were compared. Cox analyses and stratified life table analyses were used to adjust for differences in 13 variables including concurrent medical therapy. Complete follow-up was obtained in 97% of PTCA patients and 99% of medically treated patients. After correction for baseline differences in significantly predictive variables, there was no difference between the relative risk of death after PTCA vs medical therapy (RR = 0.7; p = 0.36). However, results of analysis of prospectively defined subsets showed a potential survival benefit after PTCA for patients with an ejection fraction less than 50% (RR = 0.2; p = 0.02) and also for patients with two-vessel disease (RR = 0.2; p = 0.04). For the groups as a whole there was no difference in risk of infarction after PTCA and medical therapy (RR = 0.8; p = 0.58). However, for patients with 90% to 99% LAD stenosis there was a trend toward lessened risk of infarction after PTCA (RR = 0.6; p = 0.15). No patient subset had a higher risk of death or infarction with PTCA (p less than or equal to 0.15). The likelihood of later surgery was increased after PTCA (RR = 1.5; p = 0.002). Angina, employment, and activity levels were improved after PTCA. Thus PTCA compared to medical therapy was associated with improved functional status and may decrease the risk of death and infarction in certain patient subsets. However, bypass surgery was more often performed in patients initially treated with PTCA. These data should be interpreted in light of their nonrandomized study origin and generation from different clinical sites. PMID- 2568746 TI - Post-exercise asystolic arrest in a young man without organic heart disease: utility of head-up tilt testing in guiding therapy. PMID- 2568747 TI - Hexamethonium, a forgotten drug in relation to "new" concepts in the management of heart failure. PMID- 2568748 TI - Nonthrombolytic intervention in acute myocardial infarction. AB - Alternative interventions are available for patients in whom thrombolytic therapy is inappropriate after an acute myocardial infarction. Administration of a beta blocker within the first 24 hours of the patient's admission to the coronary care unit can reduce overall morbidity and mortality within the first 7 days by about 15%. Maintenance therapy with an oral beta blocker can reduce mortality within the succeeding 3 years by about 25%. Esmolol, a unique cardioselective beta 1 adrenergic receptor blocker with a half-life of 9 minutes, can enable some patients with relative contraindications to beta blockers to nevertheless benefit from early beta-blocking therapy. It also is useful in screening patients for subsequent therapy with beta blockers. Those who tolerate the esmolol infusion can be given a long-acting beta blocker. For patients who exhibit intolerance to esmolol, the infusion can be terminated with rapid return to baseline hemodynamics. PMID- 2568749 TI - Identification of noradrenergic nerve terminals immunoreactive for neuropeptide Y and vasoactive intestinal peptide in the rat kidney. AB - Cryostat- and vibratome-cut sections of rat kidneys were singly or doubly labeled to visualize immunoreactive tyrosine hydroxylase (THI), dopamine beta-hydroxylase (DBHI), vasoactive intestinal peptide (VIPI), and neuropeptide Y (NPYI). Rats were perfusion fixed with 2-4% paraformaldehyde with or without 0.15% picric acid and rinsed in buffer for 18-48 hr. Single antigens were labeled with horseradish peroxidase in vibratome sections, whereas cryostat sections were used to label one antigen with peroxidase and another with a fluorophore in the same tissue section. A dense plexus of DBHI noradrenergic nerves innervates the renal arterial tree, and such nerves innervate the interlobar veins and renal calyx as well. Immunoreactive NPY is colocalized in most of these nerves, but some intrarenal noradrenergic nerves do not contain NPY but do contain VIP immunoreactivity. The distribution of NPYI nerves resembles that of DBHI nerves, whereas most perivascular noradrenergic nerves immunoreactive for VIP innervate selected arcuate and interlobular arteries. A small population of nonadrenergic, VIPI nerves innervates the renal calyx. PMID- 2568750 TI - Pharmacy intern intervention to reduce costs associated with histamine H2 antagonist therapy. PMID- 2568751 TI - Marked clinical difference between two sibs affected with juvenile metachromatic leukodystrophy. AB - In a child with enzymatically and histopathologically proven metachromatic leukodystrophy (MLD), the disease pursued a course typical of juvenile MLD characterized by neurological degeneration beginning at age 9 years and ending in death at age 18. A younger brother of the patient was found to have profound deficiency of arylsulfatase A in leukocytes and to excrete five- to 20-fold greater-than-normal amounts of sulfatide in the urine. He was completely free of symptoms attributable to MLD until age 16 when he developed acute cholecystitis caused by sulfatide accumulation in the gallbladder. Results of detailed neurological examination at age 21 years were normal; formal psychometric assessment showed a full-scale IQ of 105 (Wechsler). Studies on cultured skin fibroblasts from the brother showed defects in arylsulfatase A activity, measured with the use of synthetic and natural substrates, and in radiolabeled sulfatide turnover. Cellulose acetate gel electrophoresis of fibroblast extracts from the patient showed no detectable arylsulfatase A isozyme under conditions that clearly distinguished pseudo-arylsulfatase A deficiency from classical MLD. Biochemically, the patient was indistinguishable from patients with classical MLD; on the other hand, his clinical course is dramatically more benign than that of his sister who was affected with severe MLD. PMID- 2568752 TI - Restriction fragment length polymorphisms within proximal 15q and their use in molecular cytogenetics and the Prader-Willi syndrome. AB - Restriction fragment length polymorphisms (RFLPs) are described in detail for 6 DNA probes (D15S9-13, D15S18) that localize to the proximal long arm of human chromosome 15 (15q11-15q13: this report and Tantravahi et al., Am. J. Med. Genet. 33:78-87. Multiple RFLPs are detected by the probe that identifies locus D15S13, and these RFLPs are shown by genomic mapping to result from a nearby insertion or deletion of 1.8 kilobases (kb) of DNA. This set of RFLPs detected by proximal 15q probes can be used for studies on the Prader-Willi syndrome (PWS) and on mentally retarded individuals with a supernumerary inv dup(15) chromosome. Five of the polymorphic loci (D15S9-13) map to the region implicated in the cause of the PWS (15q11.2-15q12). Each of 4 families tested with these probes, as well as an additional "PWS-like" patient, was informative by RFLP analysis. The two PWS deletions studied, which occurred de novo, were inherited from the chromosome 15 provided by the father. By contrast, the 2 inv dup(15) chromosomes analyzed were of maternal origin. The use of RFLPs can also simplify the molecular determination of copy number in chromosomal aneuploidy, as exemplified by analysis of individuals with the PWS and a deletion, patients with an inv dup(15), and one patient with a more complex rearrangement involving chromosome 15. Our studies demonstrate the application of DNA probes for both molecular cytogenetic studies on this chromosome region and the development of diagnostic molecular markers to aid early clinical diagnosis of the PWS. PMID- 2568753 TI - Molecular diagnosis of the fragile X [Fra (X)] syndrome: calculation of risks based on flanking DNA markers in small phase-unknown families. AB - We studied two small, two-generation families with the fragile X [Fra (X)] syndrome. The absolute phase of the DNA markers in relation to the disease in the mother was not known in either family. We present the derivation of risks for these families using flanking markers, taking into account the uncertainty regarding maternal phase. Since the use of flanking markers fails to yield useful counseling data in the 1/3 to 1/7 of all cases where a single recombination event occurs between the two flanking markers, we calculate the probability that this method is likely to be successful or unsuccessful when prenatal diagnosis is attempted using linked RFLPs. PMID- 2568754 TI - Neural mechanisms of pancreatic polypeptide release in conscious dogs. AB - L364,718, a potent and specific antagonist for peripheral cholecystokinin (CCK) receptors, was used to determine its effect on plasma levels of pancreatic polypeptide (PP) after administration of 2-deoxy-D-glucose (2-DG, a central vagal activator) and of bethanechol (a cholinergic receptor agonist). Six conscious dogs were used in this study. Intravenous injection of 2-DG (75 mg/kg) caused significant increases in plasma levels of PP and gastrin, but there was no significant rise in plasma levels of immunoreactive and bioactive CCK. Intravenous injection of L364,718 (20 nmol/kg) significantly inhibited the PP response stimulated by 2-DG injection by approximately 60% but did not affect gastrin. Plasma levels of PP were increased dose dependently by bethanechol infusion and were not altered significantly by injections of L364,718. The results indicate that L364,718 inhibits PP response stimulated by a central vagal activator (2-DG) but not by cholinergic receptor agonist (bethanechol). This study suggests that CCK might be involved in the neural control of PP release as a neurotransmitter but probably not as a final activator of PP cells in dogs. PMID- 2568755 TI - Arteriolar smooth muscle responses are modulated by an intramural diffusion barrier. AB - Arterioles (40-80 micron diameter) were isolated from the hamster cheek pouch, cannulated at both ends, and perfused with 3-(N-morpholino)propanesulfonic acid (MOPS)-buffered physiological salt solution (PSS). The vessels were observed with an inverted microscope and video system, and arteriolar diameter was measured. Arterioles were found to be 100 times more responsive to the alpha 1-adrenoceptor agonist phenylephrine when applied to the adventitial surface than when applied to the luminal surface. In contrast, SKF 89748-A, also an alpha 1-adrenoceptor selective agonist, but with a much greater lipid solubility than phenylephrine, was equipotent from either surface of the arteriole. We hypothesized that the difference between the two drugs was due to the ability of SKF 89748-A to permeate a diffusion barrier in the arteriolar wall because of its lipid solubility. To test this hypothesis, a spectrum of antagonists with different sites of action and lipid solubilities was tested. The alpha-adrenoceptor antagonists phentolamine and benextramine and the muscarinic receptor antagonists atropine, scopolamine, and methscopolamine were all found to be more potent at blocking the action of appropriate agonists when applied to the same surface of the arteriole as the agonist than when applied to the opposite surface. Octanol water partition coefficients were measured for each of the compounds, and these were found to be highly correlated with the ratio of luminal potency to adventitial potency for each of the drugs tested. These data support the hypothesis that the endothelial cell layer in these arterioles forms a barrier to the diffusion of small, water-soluble molecules from the lumen to the smooth muscle cell layer. Such a barrier may have a significant effect on arteriolar reactivity. PMID- 2568756 TI - Effect of circulating vasopressin on arterial pressure regulation in rats. AB - It has been hypothesized that moderately increased blood levels of arginine vasopressin (AVP) contribute to the development and/or maintenance of hypertension. In this study, male Sprague-Dawley rats on a fixed 1 meq daily sodium intake received 10-day intravenous infusions of 0.2 and 2.0 ng.kg-1.min-1 AVP. The higher infusion rate was above the acute vasoconstrictor threshold for AVP administration and also produced a maximal antidiuretic effect. During chronic AVP administration, however, daily mean arterial pressure, heart rate, and body fluid composition were not changed, despite a maintained antidiuresis. To test the hypothesis that circulating AVP failed to cause hypertension as a result of sensitization of the baroreflex or a direct sympathoinhibitory effect of the peptide, additional experiments were performed in rats subjected to sinoaortic denervation (SAD) or ablation of the area postrema (APX). Infusion of AVP for 10 days into SAD or APX rats caused a sustained antidiuresis but did not change arterial pressure, heart rate, or body fluid composition. In all groups of rats, the depressor response to ganglionic blockade (20 mg/kg hexamethonium) was used to estimate the autonomic component of resting arterial pressure; no change in autonomic cardiovascular control was found using this method in any of the groups during AVP infusion. Long-term elevation of plasma AVP in rats, therefore, does not cause hypertension or significantly affect autonomic regulation of arterial pressure. PMID- 2568757 TI - Baroreceptor denervation profoundly enhances cardiovascular responses to central angiotensin II. AB - The purpose of this study was to examine the influence of arterial baroreflexes on the pressor and regional hemodynamic responses to centrally administered angiotensin II (ANG II) in the conscious rat. Fourteen days after sinoaortic baroreceptor denervation (SAD) or the equivalent sham surgery, the pressor and hindquarters vascular resistance responses to intravenous administration of ANG II were augmented in the SAD group. The pressor and vasoconstrictor responses to intracerebroventricular ANG II were also augmented after SAD; however, the SAD animals were more than 1,000-fold more sensitive than the sham group to the pressor effects of intracerebroventricular ANG II. Further experiments demonstrated that 1) the enhanced pressor response to intracerebroventricular ANG II in the baroreceptor-denervated group was due to similar increases in sympathetic outflow and vasopressin mediated vasoconstriction, 2) the increased sensitivity to central ANG II occurs as soon as 1 h after SAD, and 3) the enhanced pressor effects to intracerebroventricular ANG II also occur with intracerebroventricular hypertonic saline. We conclude that arterial baroreflexes exert a potent central inhibitory effect on the central pressor actions of ANG II that are greater than can be accounted for by the peripheral reflex arc. Finally, because of the rapid onset of the increased responsiveness to central ANG II after SAD, we propose that baroreflex buffering of central pressor stimuli may be tonically involved in circulatory control. PMID- 2568758 TI - Venoconstriction and central mobilization of blood. PMID- 2568759 TI - Interaction of CCK-8 and somatostatin-14 in control of food intake in dogs. AB - interactions of cholecystokinin COOH-terminal octapeptide (CCK-8) and somatostatin-14 (SS-14) on food intake in dogs were examined by administration of graded doses of these peptides alone and in combination. In animals fasted 19 h, SS-14 (2,000 or 20,000 pmol.kg-1.h-1) had no effect on food intake. In animals fasted 4 h, food intake was not affected by 40 or 400 pmol.kg-1.h-1 SS-14 but was significantly (P less than 0.05) increased by 20% after 4,000 pmol.kg-1.h-1 SS 14. Feeding responses to simultaneous infusions of CCK-8 (50 or 400 pmol.kg-1.h 1) and SS-14 (40, 400, or 4,000 pmol.kg-1.h-1) were determined in animals fasted 4 h. Given alone, the high dose of CCK-8 (400 pmol.kg-1.h-1) significantly (P less than 0.01) depressed food intake by 55%. This effect was blocked by all doses of SS-14. In the absence of CCK-8, SS-14 had no effect except at the highest dose (4,000 pmol.kg-1.h-1), which significantly (P less than 0.01) stimulated food intake by 57%. This effect was blocked by both doses of CCK-8. Simultaneous infusion of lower doses of SS-14 (40 and 400 pmol.kg-1.h-1) and CCK 8 (50 pmol.kg-1.h-1) had no effect on food intake. These results suggest that plasma levels of CCK and SS-14 after a meal are not sufficient alone or in combination to produce satiety. PMID- 2568760 TI - Na-dependent L-glutamate transport by eel intestinal BBMV: role of K+ and Cl-. AB - L-[3H]glutamate uptake into eel (Anguilla anguilla) intestinal brush-border membrane vesicles (BBMV) was a sigmoidal function of extravesicular Na, suggesting that two or more cations accompanied the amino acid during transport. L-[3H]glutamate influx illustrated the following kinetic constants: apparent membrane binding affinity (Kapp) = 0.80 +/- 0.12 mM; influx velocity (Jmax) = 2.61 +/- 0.31 nmol.mg protein-1.min-1; and permeability coefficient (P) = 0.65 +/ 0.10 microliters.mg protein-1. min-1. Results from the imposition of diffusion potentials across vesicle membranes using K-valinomycin or H-carbonyl-cyanide p chloromethoxyphenylhydrazone suggested that Na-dependent L-glutamate transport was sensitive to transmembrane electrical potential difference. Extravesicular aspartate was a competitive inhibitor of L-[3H]glutamate influx [inhibitory constant (Ki) = 0.28 +/- 0.04 mM]. Intravesicular K and extravesicular Cl ions enhanced maximal amino acid influx and transient L-glutamate accumulation against a concentration gradient (overshoot). Intravesicular K reduced the Kapp of the membrane to L-glutamate, whereas extravesicular Cl increased L-glutamate Jmax. A model for L-[3H]glutamate transport is suggested involving the cotransport of at least two Na and one L-glutamate that is activated by one intravesicular K ion and at least two extravesicular Cl ions. PMID- 2568762 TI - Alprazolam augmentation of neuroleptics in schizophrenia. PMID- 2568761 TI - Axon destruction and adrenergic systems mediate pressor responses after AV3V lesions. AB - These studies investigated the neural tissue and peripheral mechanism mediating the transient pressor response following electrolytic ablation of the periventricular tissue surrounding the anteroventral third ventricle (AV3V) of the rat. Arterial blood pressure was monitored in conscious animals for 2 h following either microinjection of kainic acid (AV3V-KA) or electrolytic lesions (AV3V-X) in the AV3V region or control procedures (Cont). Blood pressure did not change in AV3V-KA (4 +/- 3 mmHg) or Cont rats but significantly increased in AV3V X animals (20 +/- 4 mmHg). The pressor response following AV3V-X was not altered by pretreatment with MK-422 (converting-enzyme inhibitor), TMe-AVP (vasopressin antagonist), or hexamethonium (ganglionic blocker). However, intravenous administration of prazosin (alpha-adrenergic blocker) or bilateral adrenalectomy abolished the increase in blood pressure. Furthermore, plasma concentrations of norepinephrine were significantly higher in AV3V-X rats (1,125 +/- 150 pg/ml) compared with Cont animals (322 +/- 83 pg/ml) following treatment. These data indicate that the acute hypertensive response following AV3V-X is caused by the destruction of fibers of passage and results from circulating catecholamines of adrenal origin. PMID- 2568763 TI - Neurochemical abnormalities of anorexia nervosa and bulimia nervosa. AB - The authors review the research on anorexia nervosa and bulimia nervosa, emphasizing the neurotransmitters and neuromodulators that regulate eating behavior. Anorexia nervosa is associated with changes in the noradrenergic, serotonergic, and opioid systems; bulimia nervosa is accompanied by marked alterations in serotonin and norepinephrine activity. These neurochemical changes may perpetuate pathological eating behavior and may be responsible for several associated psychiatric symptoms, including anxiety and depression. The authors also summarize studies of several drugs that are used in the treatment of eating disorders and are known to modify neurotransmitter activity. Understanding the neurochemistry of eating disorders seems crucial for the rational development of both psychopharmacological and behavioral treatments. PMID- 2568764 TI - Tuberous sclerosis: aberrant proline sensitivity of skin fibroblasts. AB - Cultured skin fibroblasts from patients with tuberous sclerosis were analyzed as to their sensitivities to proline, glutamic acid and arginine, and their analogues by means of a cell growth study. A difference was observed between tuberous sclerosis and normal cells when proline was added. Skin fibroblasts derived from normal-appearing skin of tuberous sclerosis patients exhibited sensitivity to proline. Skin fibroblasts derived from tumorous skin of tuberous sclerosis patients exhibited resistance to proline in the cell growth study and was found to be rather sensitive to proline in a colony formation study. This aberrant sensitivity to proline of tuberous sclerosis cells may indicate abnormal proline metabolism in tuberous sclerosis patients. PMID- 2568765 TI - Alcohol withdrawal syndrome: clinical and hormonal responses to alpha 2 adrenergic agonist treatment. AB - Lofexidine (L) was compared to placebo (P) in a random double blind placebo controlled study of 23 healthy alcoholics in withdrawal. In serial alcohol withdrawal syndrome (AWS) scales, P greater than L for first 18 hr. The major changes in AWS were in blood pressure and pulse, while restlessness or diaphoresis did not show faster normalization with L vs. P. Other ratings of withdrawal intensities and craving by patients were similar in both P and L. There were no differences in the rate of patient completion, or appearance of hallucinations by group. Plasma catecholamines were in the high ranges, similar in both groups at 0 and 3 hr, but after 5 hr P greater than L for norepinephrine (but not epinephrine or dopamine). The data suggest L has pharmacological effects on some objective, catecholamine related, components of alcohol withdrawal, with little effect on others. L type treatment of alcoholics in withdrawal is promising, modestly effective and warrants further study. PMID- 2568766 TI - Laboratory markers as compared to drinking measures before and after inpatient treatment for alcoholism. AB - A study group, consisting of 60 male and 13 female alcohol-dependent patients, participated in an inpatient treatment for alcoholism. They were interviewed about their alcohol use before the treatment period and bimonthly for 8 months after it. It was found that the correlations between laboratory markers and average daily alcohol intake are very low (range, -0.01-0.23) in alcoholics with high alcoholic intake level (before the treatment period). It was also found that the correlations between the laboratory tests and the drinking measures after the initial assessment undergo large changes. The correlation between GGT and average daily alcohol intake before the treatment period was 0.07 (p = NS) and 2 months after the treatment period 0.55 (p less than 0.001). The long period of abstinence, the change to a lower level of alcohol consumption, and improved accuracy of reporting alcohol intake were probably causative in this change. GGT proved to have the highest correlations with the drinking measures during the follow-up and MCV before the treatment period. The frequency measure, number of drinking days, gave consistently higher correlations with the laboratory markers than the measure of drinking amount, average daily alcohol intake. Laboratory markers, GGT and MCV, are well suited for outcome evaluation of drinking behavior of alcoholics after inpatient treatment for alcoholism or other comparative periods of abstinence. PMID- 2568767 TI - Dose-response relationships for edrophonium and neostigmine as antagonists of atracurium and vecuronium neuromuscular blockade. AB - To determine the potencies of edrophonium and neostigmine as antagonists of nondepolarizing neuromuscular blockade produced by atracurium and vecuronium, dose-response curves were constructed for both antagonists when given at 10% spontaneous recovery of first twitch height. Ninety ASA physical status 1 and 2 adults were given either 0.4 mg/kg atracurium or 0.08 mg/kg vecuronium during thiopental-nitrous oxide-enflurane anesthesia. Train-of-four stimulation was applied to the ulnar nerve every 12 s, and the force of contraction of the adductor pollicis muscle was recorded. When spontaneous recovery of first twitch height reached 10% of its initial control value, edrophonium (0.1, 0.2, 0.4, or 1 mg/kg) or neostigmine (0.005, 0.01, 0.02, or 0.05 mg/kg) was administered by random allocation. Neuromuscular function in another ten subjects was allowed to recover spontaneously. Assisted recovery was defined as actual recovery minus mean spontaneous recovery observed in patients who were not given antagonists. First twitch recovery was initially more rapid when vecuronium was antagonized compared with atracurium, but no difference was detected after 10 min. At 10 min the neostigmine ED80 was 0.022 +/- 0.003 (SEM) mg/kg after atracurium and 0.024 +/- 0.003 mg/kg after vecuronium. The edrophonium ED80 was 0.44 +/- 0.11 mg/kg with atracurium and 0.46 +/- 0.12 mg/kg with vecuronium, giving a neostigmine:edrophonium potency ratio of 20. Atracurium train-of-four fade could be antagonized more easily with edrophonium, whereas that of vecuronium was more easily antagonized by neostigmine. It is concluded that edrophonium and neostigmine are not equally effective against atracurium and vecuronium. PMID- 2568768 TI - Atracurium, vecuronium, and pancuronium do not alter the minimum alveolar concentration of halothane in humans. AB - The authors studied 64 unpremedicated, healthy surgical patients, aged 42 +/- 14 yr, to determine the effects of atracurium, vecuronium, and pancuronium on the minimum alveolar concentration (MAC) of halothane. Anesthesia was induced using halothane/nitrous oxide/oxygen via a mask without the administration of other drugs. Nitrous oxide was discontinued, the trachea was intubated without prior administration of neuromuscular blocking drugs, and anesthesia was maintained with halothane in oxygen. Participating patients were assigned to one of five groups: 1) no neuromuscular blocking drug (control group, n = 9); 2) atracurium 0.5 mg/kg (n = 10); 3) atracurium 1.0 mg/kg (n = 15); 4) vecuronium 0.1 mg/kg (n = 20); or, 5) pancuronium 0.1 mg/kg (n = 10). Tourniquets, inflated to 300 mmHg immediately before iv administration of neuromuscular blocking drug and 15-30 min prior to skin incision, were used to isolate extremities from circulating neuromuscular blocking drug in all patients. A positive response to stimulation was defined as movement of at least one extremity occurring distal to the tourniquet within 1 min following skin incision. The first patients in the control and atracurium groups were studied at an end-tidal halothane concentration of 0.95%. The first patient in the pancuronium group was studied at a halothane concentration of 0.75%, and the first patient in the vecuronium group at 0.70%. Subsequent patients were studied at end-tidal halothane concentrations 0.10% above or below that of the preceding patient, depending on the presence or absence of movement with skin incision. Control MAC for halothane was 0.74% +/- 0.09% (mean +/- SEM).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568769 TI - Dexmedetomidine produces a hypnotic-anesthetic action in rats via activation of central alpha-2 adrenoceptors. AB - Dexmedetomidine, a highly selective and potent alpha-2 adrenoceptor agonist, reduces halothane anesthetic requirements by over 90% in rats. The present study examined whether dexmedetomidine produces a hypnotic-anesthetic action in rats. Dexmedetomidine induced a hypnotic-anesthetic state in rats characterized by loss of righting reflex at doses greater than or equal to 0.1 mg/kg. This response was dose-dependent between 0.1 and 3 mg/kg. Alpha-2 adrenoceptor antagonists that cross the blood-brain barrier (antipamezole and idazoxan) decreased the hypnotic anesthetic action of dexmedetomidine in a dose-dependent fashion. In contrast, the alpha-2 antagonist, L-659,066, which does not penetrate into the CNS did not affect dexmedetomidine-induced hypnosis. Antagonists for the other adrenoceptors not only failed to reduce the hypnotic-anesthetic action of dexmedetomidine but in some cases even potentiated this effect. Thus, prazosin, an alpha-1 adrenoceptor antagonist, significantly enhanced the hypnotic-anesthetic property of dexmedetomidine. Antagonists with beta-2 receptor blocking properties also enhanced dexmedetomidine-induced hypnosis. Selective beta-1 receptor antagonists did not affect the hypnotic action of dexmedetomidine. These results suggest that dexmedetomidine produces a hypnotic-anesthetic action in rats via activation of central alpha-2 adrenoceptors. PMID- 2568770 TI - Autocytotoxic activity of lymphokine-activated killer cells: characterization of effector cells and susceptible targets. AB - The specificities and surface markers of murine autocytotoxic cells induced by in vitro culture with interleukin 2 (IL2) were studied. Culturing murine spleen cells with recombinant human IL2 resulted in the generation of cytotoxic cells which killed syngeneic lymphoblasts and syngeneic activated macrophages (M phi). Both lectins and protein antigens were capable of inducing lymphoblasts recognized by lymphokine-activated killer (LAK) cells. B-lymphoblasts as well as T-lymphoblasts were sensitive to lysis by these effector cells. In addition, peritoneal M phi activated in vivo with Bacille Calmette-Guerin (BCB), Corynebacterium parvum (C. parvum), thioglycollate (TG) or lipopolysaccharide (LPS) were shown to be susceptible to lysis by LAK cells. In contrast, neither unstimulated T cells nor resident peritoneal M phi were sensitive to lysis by LAK cells, suggesting that normal cells have to be activated in order to be sensitive to lysis by these effector cells. Surface marker analysis indicated that majority of effector cells which killed syngeneic lymphoblasts and activated M phi were Thy1+, asialo GM1+, L3T4-, Ly2-. PMID- 2568771 TI - Amplification of oncogenes in lung carcinoma grafted in nude mice. AB - Seven lung carcinomas were grafted on nude mice and continuously propagated as in vivo models on which the amplification of 9 oncogenes (N-myc, v-erb A, v-abl, v sis, c-myc, c-myb, v-Ha-ras, c-Kiras, and v-scr) was studied by Southern blot hybridization. Only c-myc was amplified (20 copies) in an adenocarcinoma. The presence of 2 bands at 9 kb and 6.6 kb in addition to the normal 12.7 kb in EcoR1 digested DNAs suggested a polymorphism of the c-myc gene in this tumor. The other 8 oncogenes were not amplified in this tumor. The 5 small cell lung carcinomas of this study did not show any amplification of any of the 9 oncogenes tested. PMID- 2568773 TI - Recent developments in nonionic contrast media. PMID- 2568772 TI - Influence of fluoroquinolones on expression and function of P fimbriae in uropathogenic Escherichia coli. AB - P fimbriae are the major adhesins mediating attachment of pyelonephritogenic Escherichia coli to urinary tract tissues, and they therefore constitute a recognized virulence factor. In this work, the effect of fluoroquinolones on P fimbria expression and function in E. coli SS142 and C1212 was assessed. Ciprofloxacin, fleroxacin, and norfloxacin were compared with their precursor nalidixic acid and with trimethoprim in sublethal concentrations ranging from 1/32 to 1/4 of the MIC. Fimbria function was assessed in a standard hemagglutination assay and in a parallel hemagglutination inhibition assay in which the tier of antifimbrial antiserum necessary to inhibit hemagglutination by SS142 was determined. Adhesion of antibiotic-exposed bacteria to human uroma T24 cells in suspension was also measured. Fimbria production was quantitated in an inhibition enzyme-linked immunosorbent assay. Trimethoprim produced a dose dependent decrease of three to four hemagglutination titers for both strains and a decline in the antiserum titer from 1:16 (control) to 1:128 (1/4 MIC) for E. coli SS142. Adherence exhibited similar decrements from 130 +/- 28 (control) to 16 +/- 3 (1/4 MIC) and from 83 +/- 19 (control) to 30 +/- 11 (1/4 MIC) E. coli cells per uroepithelial cell (mean +/- standard error) for SS142 and C1212, respectively (P less than 0.015). By enzyme-linked immunosorbent assay, the inhibition following exposure decreased in a dose-dependent manner from 31% (control) to 8% (1/4 MIC). By contrast, none of the quinolones produced significant changes in the parameters assessed above. At sublethal concentrations, trimethoprim decreased fimbria production. Following exposure to fluoroquinolones, however, E. coli expressed morphologically and functionally intact P fimbriae. PMID- 2568774 TI - Renal excretion of iopromide and iopamidol after intravenous administration in digital subtraction angiography. AB - After injection of iopromide or iopamidol into a central vein in 40 patients ages 57 +/- 14 years and 57 +/- 11 years, respectively, with normal renal function, iodine concentration and total iodine excretion were determined in pooled urine over 0 to 2 hours and 2 to 24 hours after injection. Iopromide excretion of 46.4 +/- 9.8% of the dose during the first 2 hours after injection was significantly (p less than 0.05) higher than that of iopamidol, which amounted to 41.4 +/- 10.5% of the dose. This is presumably due to the fact that iopromide has lower protein binding than iopamidol. Excretion of the two nonionic contrast media up to 2 hours after injection decreased constantly with increasing age. In the case of iopromide, a mean excretion of 55% of the dose in a patient 25 years old decreased to about 40% for a patient 90 years of age (p less than 0.05). The clinical relevance of the observed differences and correlations is discussed. PMID- 2568776 TI - Clinical experience with iohexol versus iopromide in excretory urography. AB - In a controlled double-blind study the quality of the contrast produced by iopromide and iohexol was compared in two groups of 100 patients by excretory urography. The aim was to investigate whether iopromide or iohexol has any diagnostic advantages for the opacification of the kidneys, the renal pelves, and the ureters. The results show that there is no difference in contrast quality between the two substances (chi-square, p greater than 0.1). Moreover there is no difference in the frequency of concomitant symptoms (chi-square, p greater than 0.1). Both contrast media show the well-known excellent tolerance with only 1% side effects and the good contrast quality of the nonionic contrast media. PMID- 2568775 TI - Influence of contrast media osmolality on intravenous urographic quality. AB - A clinical study was designed to determine whether the lower solute load after intravenous injection of nonionic contrast medium (CM) produced significant changes in nephrographic and pyelographic quality. For equal iodine doses (300 mg I/kg), the lower solute load of nonionic CM produced better nephrogram and pyelogram scores than ionic CM or nonionic CM with mannitol, both of which had higher solute loads. The higher scores associated with nonionic CM were statistically significant for the pyelogram phase but not for the nephrogram phase. These observations were matched by lesser changes in urinary osmolality and higher urinary iodine concentrations with the nonionic CM. PMID- 2568778 TI - Urography with monomeric and dimeric nonionic contrast media: comparative, randomized, double-blind study of iotrolan 280 and iopromide 300. AB - In a double-blind comparative study between the nonionic, dimeric iotrolan and the nonionic, monomeric iopromide the urographic image quality in the dose 300 mg I/kg body weight is better after iopromide up to 20 minutes after the injection. This result appears to be in contradiction to the results of the animal experimental studies. Possible reasons are discussed. PMID- 2568777 TI - Iopromide dosage and urographic image quality: is there an optimal dose? AB - The influence of increasing doses of contrast medium (CM; 0.75, 1.0, 1.5, and 2.0 ml iopromide/kg body weight; 300 mg I/ml) was examined in a randomized, double blind study. An increase of the dosage resulted in a statistically significant improvement of the quality of the radiographic visualization. This was most pronounced with an increase from 0.75 to 1.0 ml/kg body weight and least pronounced with the increase from 1.5 to 2.0 ml/kg body weight. This improvement was not only visible in the overall quality, but in each part of the urinary tract, the calyces, the pelves, the ureters, and most especially at the parenchyma. Due to economic considerations at the present, a dose of 1.0 ml/kg (300 mg I/ml) is viewed as adequate. Doses below this level should be avoided due to the poorer image quality that results. From a purely medical point of view, the injection of 1.5 ml of nonionic CM/kg body weight is considered optimal. PMID- 2568779 TI - Elimination of the nonionic contrast medium iopromide in end-stage renal failure by hemodialysis. AB - The increasing life-expectancy of patients with end-stage renal failure has led to a significant increase of diagnostic and interventional radiologic measures. The contrast media used in these procedures, however, are excreted only very slowly, due to the renal failure. We have therefore studied the elimination of the nonionic x-ray contrast medium iopromide (Ultravist) during dialysis in nine anuric patients with end-stage renal failure. All of the patients received 100 ml iopromide (30 g iodine) for the visualization of their Cimino-Brescia shunt. They were then dialyzed 30 minutes after the injection. The plasma iodine concentration declined from 1.7 to 0.7 mg I/ml plasma during the 3-hour dialysis treatment. The iopromide clearance was a constant 80 ml/min. This is the equivalent of a relative dialysance of 65% of the creatinine clearance. There were no incidences of side effects requiring therapy in this high-risk group of patients. The quality of the image was good in all cases. Iopromide proved itself to be an x-ray contrast medium with a low rate of side effects, which can be effectively eliminated from the body by means of conventional dialysis. PMID- 2568780 TI - Tolerance to iotrolan after subarachnoid injection in animals. AB - Neural tolerance after intracisternal administration of iotrolan was compared with that after iohexol, iopamidol, and metrizamide in mice, rats, and guinea pigs. Around the level of the ED50 (approximately two to four times the human dose) tolerance to iotrolan appeared to be much better than tolerance to the other agents. A study in rabbits comparing iotrolan with iohexol produced approximately the same result. High doses of iotrolan, iohexol, and iopamidol were almost equally well tolerated by rats, as were iotrolan and iohexol by rabbits. Tolerance to metrizamide by rats and to iopamidol by guinea pigs was vastly inferior. In support of this very good general tolerance histologic examinations of the spinal tract and of the brain did not reveal any substance related changes in beagles after lumbar administration of a high dose. An investigation in rats using mannitol and sorbitol formulations with differing osmotic pressures indicates that contrast tolerance is influenced primarily by the chemotoxicity and not by increased osmotic pressure. As shown by the results of the preclinical investigations, iotrolan should be ideal for use in myelography and also appears highly suitable for the examination of other body cavities. PMID- 2568781 TI - A possible mechanism for the neural adverse reactions caused by metrizamide. AB - Causality of the metrizamide-induced neural adverse effects was explored among the effects on behavior, electroencephalogram (EEG), and brain glucose utilization in rats. Iotrolan, a new myelographic contrast agent, was used as a reference substance throughout the study. Supracortical subarachnoidal administration of metrizamide caused, within a few minutes, symptoms of sedation and anxiety, which were accompanied by appearance of slow wave or flattening in EEG not only of the cortex, but also of the regions of the hippocampus and thalamus. The rates of local cerebral glucose utilization (LCGU) in a wide range of the brain, measured by using 3H-2-deoxyglucose, were also altered significantly. Despite a limited distribution of metrizamide in the lateral region of the cortex, LCGU was suppressed significantly in the administered side of the parietal cortex, thalamus, subthalamic nucleus, medial geniculate body, and mammillary body and increased in the regions of hippocampus, caudate-putamen, and globus pallidus. It is concluded that, rather than inhibiting the hexokinase reaction in the brain cell, metrizamide appears to cause reduction of a net glucose transport into the cell and that this direct effect on the cortex is amplified and propagated, via neurotransmission, to the regions of the diencephalon and midbrain, causing secondarily various types of disturbance in the mental and motor functions. Iotrolan was proved to lack any biologic activity that may relate to the neural adverse effect observed with metrizamide. PMID- 2568782 TI - Diskography with iotrolan before chemonucleolysis with chymopapain. AB - The neuroradiologic changes after chemonucleolysis were studied in animal experiments conducted to establish whether there is any interaction between chymopapain and the contrast medium iotrolan. Twenty canine disks were examined by diskography with iotrolan 300 mg I/ml before nucleolysis with chymopapain. Twenty disks were subjected only to nucleolysis. For control purposes, another 10 disks were examined by diskography without the administration of chymopapain. The neuroradiologic follow-up study with conventional radiography, computed tomography and magnetic resonance imaging of the lumbar vertebral column revealed typical signs of the effect of chymopapain in all nucleolysed disks, regardless of whether diskography had been performed before chemonucleolysis or not. On the basis of our results, there is no need to fear an interaction between the new dimeric, nonionic contrast medium iotrolan and the substance chymopapain used for chemonucleolysis. PMID- 2568783 TI - Tolerance of iotrolan 190 and cerebrospinal fluid-diluted iotrolan 300 at the same iodine concentration in lumbar myelography. AB - Iotrolan, the radiologically effective ingredient of the contrast medium Isovist, has so little osmotic activity that only solutions of about 300 mg I/ml are isotonic to cerebrospinal fluid (CSF). The osmolality of the contrast medium at lower iodine concentrations has to be increased through additives. Theoretically, iotrolan 300 diluted with CSF is the most physiologic. Therefore, the commercial preparation iotrolan 190 (adaptation of the osmolality through the addition of saline and sodium bicarbonate) was tested in a randomized, single-blind study against a diluted contrast medium solution, iotrolan 190 (produced from iotrolan 300). The dilution was performed with the patient's own CSF. The test was carried out in lumbar myelography in two groups of 50 patients each. There were neither statistical nor tendency differences between the two preparations in either the opacification or the tolerance. Therefore, there is no suggestion that a contrast medium solution prepared with the patient's own CSF is better tolerated. PMID- 2568784 TI - Iotrolan versus iopamidol: a controlled, multicenter, double-blind study of lumbar and direct cervical myelography. AB - Iotrolan is a new nonionic, dimeric contrast medium that is distinguished by its special physicochemical and low chemotoxic properties. It displays near isotonicity with blood and cerebrospinal fluid in all clinically used concentrations (190, 240, and 300 mg I/ml). The controlled, double-blind study of iotrolan versus iopamidol presented here shows that iotrolan is significantly superior to the monomeric contrast medium in lumbar and direct cervical myelography. Use of the dimeric contrast medium reduced not only the incidence of side effects, but also their severity and duration. The chemotoxicity of contrast media commonly used was significantly less with iotrolan in all neuroradiologic diagnostic studies of the spine. The diagnostic quality of lumbar and direct cervical myelography with iotrolan was high. In this randomized, double-blind study versus iopamidol, iotrolan proved to be the safer and better myelographic agent for contrast studies of the spinal canal. PMID- 2568785 TI - Iotrolan, a nonionic dimeric contrast medium in myelography. AB - As part of a clinical Phase III study of the nonionic, dimeric contrast medium iotrolan, differences in opacification, tolerance, and effects on the electroencephalogram after myelography were examined in a double-blind comparison with iohexol. Two groups of 25 patients each received either 10 ml iotrolan 240 mg I/ml or 10 ml iohexol 240 mg I/ml for lumbar myelography, and two groups of 50 patients each received either 10 ml iotrolan 300 mg I/ml or 10 ml iohexol 300 mg I/ml for lumbar, thoracic or cervical myelography. The overall assessment of the opacification in the subarachnoid space, which was moderate in only one case of each group and otherwise good, showed no differences between the preparations. The tolerance of both concentrations of iotrolan proved to be significantly better than those of iohexol: at a significance level in the chi-square test of p less than 0.001 in a simple size of 25 patients per preparation with 240 mg I/ml and at a significance level of p +/- 0.05 in a sample size of 50 patients per preparation with 300 mg I/ml. PMID- 2568786 TI - Osmolality of nonionic contrast media. AB - Solutions of different low osmolar contrast media (CM) obviously show clinically relevant differences in the osmolality despite equal iodine concentrations and similar molecular structure. To obtain precise and comparable data, the osmolality of five batches (usually) each of contrast media, iopamidol, iohexol, iopromide, and ioxaglate-all preparations commercially available-were measured by means of the vapor pressure method. The osmolality of the solutions of sodium meglumine ioxaglate with the same iodine concentration is lower than that of the nonionic CM examined. Iopromide showed the lowest osmolality and iohexol the highest value of the nonionic preparations. The differences are statistically significant as a rule. They are attributed to a varying association and hydration of the CM molecules in the solution. PMID- 2568787 TI - Comparative clinical study with iotrolan versus metrizamide in lumbar myelography. AB - In order to examine the radiopacity, safety, and usefulness of iotrolan, a new dimeric, nonionic, water-soluble contrast medium for myelography, a multicenter clinical study was performed in lumbar myelography using metrizamide as the reference drug. Both media were injected at a strength of 190 mg I/ml in 90 patients each. Both contrast media showed good opacification; there was no significant difference between the two media. The incidence and severity of side effects were significantly lower with iotrolan than those with metrizamide. No severe side effects, such as convulsion and shock, were observed with either medium. PMID- 2568788 TI - Clinical trials with iotrolan in myelographic and computed tomography applications in the United States. AB - Iotrolan, a new nonionic contrast agent, was studied in the United States using 190 mg I/ml, 240 mg I/ml, and 300 mg I/ml concentrations. Procedures performed were lumbar, thoracic, cervical, and total columnar myelography and computed tomography applications. Two multicenter double-blind studies (iotrolan versus metrizamide in 190 mg I/ml and 240 mg I/ml concentrations) and five open-label studies were conducted in a total of 863 patients (569 iotrolan, 294 metrizamide). Iotrolan and metrizamide both demonstrated diagnostic efficacy. Iotrolan demonstrated better patient tolerance in comparison with metrizamide. PMID- 2568789 TI - Neural tolerance of iotrolan 300 in ascending cervical myelography: results of a multicenter study. AB - This study examined the opacification, dose, and tolerance of iotrolan 300 on 231 patients in ascending cervical myelography. The contrast was rated good in 188 (81.4%) of the cases and satisfactory in 40 (17.3%) of the cases. The contrast was poor in only three (1.3%) cases. In 152 patients a dose of 10 ml or less of iotrolan 300 was administered. A good contrast quality was obtained in 84.2% of all examinations. From a total of 231 patients, 146 exhibited no concomitant effects. The intensity of the headache and neck ache was recorded by the patients themselves by means of an analog scale. The frequency and degree of the postmyelographic complaints did not increase with higher doses, i.e., they were not dose dependent. Neurologic irritation, in the form of radicular symptoms, appeared in only 2 of 231 examinations. These data demonstrate that iotrolan 300 is excellent for use in ascending cervical myelography. PMID- 2568790 TI - Indirect lymphography with iotrolan. AB - Iotrolan was used in concentrations of 240 and 300 mg I/ml for indirect lymphography in a total of 70 patients, and permitted a diagnostic evaluation of the peripheral lymphatics in all cases. The contrast quality was found to correlate with the concentration used. The concentration of 300 mg I/ml Iotrolan was judged to provide the better contrast density and definition. Side effects were limited to transient local erythema and pain during the infusion in one patient and a protracted reaction resembling serum sickness in another patient (Iotrolan 240 mg I/ml). Because of its excellent tolerance, Iotrolan is particularly suitable after subepidermal infusion for indirect lymphography, during which the local lymph drainage of cutaneous regions of interest can be evaluated. PMID- 2568791 TI - Arthrography with iotrolan: double-blind comparison between nonionic, monomeric (iohexol 300) and nonionic, dimeric (iotrolan 300) contrast media. AB - Arthrography of the shoulder and knee joints is optimized by the new nonionic, dimer contrast medium iotrolan 300 in comparison to the nonionic, monomer iohexol 300. Detail recognition increases and is longer lasting. The contrast density remains good for more than 40 minutes, thus making repeated examinations due to diagnostic uncertainty superfluous. PMID- 2568792 TI - Hysterosalpingography with a new contrast medium. AB - This article reports on the use of iotrolan--a new contrast medium and the first dimeric, nonionic, hexaiodinated, water-soluble preparation--in hysterosalpingography. This randomized, double-blind study compared an ionic and a nonionic monomeric preparation. The better results were achieved with iotrolan. Its unusual lack of local irritation and its high contrast quality, especially definition of contrast, merit particular mention. PMID- 2568793 TI - Gastrointestinal diagnosis with iotrolan: experience in pediatric radiology. AB - The use of low-osmolar, nonionic, monomeric contrast media has made gastrointestinal diagnosis with water-soluble contrast media less hazardous with a greater diagnostic yield. Since, however, even this group of compounds is hypertonic in comparison with blood, water is still drawn from the body into the bowel, resulting in water and electrolyte shifts and distal dilution of the contrast material with deterioration of the image quality. The use of iotrolan, which is isotonic in comparison with blood, therefore, appears to offer advantages for selected groups of patients. The superiority of the contrast medium iotrolan over iohexol was demonstrated in a double-blind study. A further 72 examinations in an open-ended study confirmed this result. PMID- 2568794 TI - Difficulties of risk determination from the use of new contrast media. AB - The difficulties with comparing data on the risks involved in the use of conventional contrast media and new low-osmolar contrast media are presented. These difficulties are a result of the necessary size of the groups to be compared and the problems of obtaining data. Prospective studies, carried out by a single researcher, probably have the greatest reliability. In multicentered studies with a larger number of participants, a number of factors must be reckoned with. The assessment of the same reaction can vary considerably, depending on the initial illness and interest of the patient or physician. This will, of course, influence the collection of the data. Due to the fact that complications and side effects are more frequent than deaths, the reduction of the first two through the use of new contrast media is statistically easier to ascertain. It can be assumed from the studies performed thus far that the risk of mortality is also reduced. Definite statistical guarantees do not as yet exist, and it is possible that they will never be produced. PMID- 2568795 TI - Nonionic contrast media in gastrointestinal studies in newborns and small infants. AB - Since January 1983, 431 patients were examined by oral or rectal application of nonionic contrast medium for gastrointestinal studies. The contrast medium was uniformly well tolerated, even by severely ill preterm babies and infants. We present a review of our experience and a list of indications for the use of nonionic contrast media in gastrointestinal studies. In addition, the urinary iodine concentration of eight patients was examined after oral application of nonionic contrast medium in order to evaluate the resorption of the substance. PMID- 2568796 TI - Tissue distribution and excretion of iotrolan after intravenous and suboccipital injections in animals. AB - The fate of iotrolan in the body is determined by its physicochemical characteristics and biologic inertness. Iotrolan is distributed in the extracellular space after intravenous or intracisternal injections. No absorption occurred after oral administration. Iotrolan is eliminated almost exclusively by renal excretion in an unmetabolized form. The results found are very similar to those published for other x-ray contrast agents used in uroangiography. PMID- 2568797 TI - Comparative assays of the chemotoxicity of iodinated radiographic contrast media based on impairment of fibrin antithrombin activity. AB - Radiographic iodized contrast media impair the antithrombin effect of the fibrin clot. The extent of the impairment correlates with dosage and lipophilicity. Normally, 90% of the thrombin activity of 2.3 IU/ml are inactivated by a fibrin clot generated by 2 mg/ml fibrinogen. The effect is due to contrast media-induced protein denaturation with subsequent impairment of thrombin binding to the fibrin clot. Two hepatogenic, two ionic, nephrotopic, and four nonionic contrast media were assayed by this test system at different concentrations. Clear-cut differences were found between the three main groups. Smaller, yet statistically significant, differences could be established between ioglicate and diatrizoate and between iosimide, iopromide or iohexol and iotrolan. No differences were found between iotroxate and iodoxamate. The results are in concordance with the partition coefficients (water/n-butanol) of these contrast media. The assay sensitivity is suitable as a screening system of the chemotoxicity of contrast media. It is easily performed, inexpensive, and assays the clinically important contrast media interaction with fibrinogen. PMID- 2568798 TI - Pharmacokinetics of iotrolan after intravenous injection into healthy volunteers. AB - In man Iotrolan is distributed in the extravascular space after intravenous injection with a half-life of about 11 minutes, the elimination half-life being about 108 minutes. After intravenous injection 99.2 +/- 1.8% of the dose was found in the urine and 0.5 +/- 0.1% in the feces within 5 days. No metabolization occurred in man and the pharmacokinetic behavior of iotrolan is comparable to that of known nonionic contrast media (e.g., iohexol, iopromide) (7, 9, 13). PMID- 2568799 TI - Whole body autoradiographic distribution studies on nonionic x-ray contrast agents in pregnant rats. AB - The time course of the distribution of radioactive label in organs and tissues was investigated after intravenous administration of iotrolan, iopamidol, iopromide, and iohexol in a concentration of 60 mg I/ml (2.2 MBq 125I/mg I) as aqueous solution to pregnant rats (18th day after conception) by a whole-body autoradiographic technique. All contrast agents were rapidly distributed within the organism, showing equal distribution patterns of radioactivity, and were rapidly excreted, mainly by the kidney. Passage of radioactivity across the blood brain and placental barriers could not clearly be shown. No specific and long lasting retention of radioactivity could be seen in any organ or tissue with the exception of the thyroid. PMID- 2568800 TI - Pharmacokinetics of iohexol, iopamidol, iopromide, and iosimide compared with meglumine diatrizoate. AB - Four nonionic contrast media (iohexol, iopamidol, iopromide, and iosimide) are compared in this clinical study in their pharmacokinetic behavior with an ionic reference preparation (meglumine diatrizoate). At a dose of 1 ml of contrast medium per kilogram of body weight with approximately the same iodine content, virtually no differences could be established in the pharmacokinetic behavior. The osmotic diuresis of the ionic substance, compared with that of the nonionic preparations--implies an increased osmotic diuresis, thus, a lower maximal iodine concentration in the urine. The elderly patients included in this study have a reduced glomerular filtration rate, which in turn implies a prolonged half-life in the blood and a retarded renal elimination. PMID- 2568801 TI - Plasma level and renal excretion of iotrolan after lumbar injection in patients. AB - Ten milliliters of iotrolan containing 240 mg I/ml were injected into the lower lumbar subarachnoid space of ten patients. Plasma level and urinary and fecal excretion were monitored for 3 days after injection. The mean iodine concentration in the plasma of about 50 micrograms/ml (6% of the dose given in the total plasma volume) peaked at 1 to 2 hours after injection. After reaching the maximum values, the iotrolan concentration in the plasma decreased with a half-life of about 4 hours (median value). About 90% of the injected dose was found in the urine and about 1% was detected in the feces. PMID- 2568802 TI - Left ventricular after load as a result of levocardiography with contrast media of various osmolality. AB - Contrast media have been successfully used as a diagnostic aid in radiology for decades. It has, however, been necessary to accept certain, in some cases unexplained, undesirable side effects because of their physicochemical properties. The conventional preparations available have for more than 40 years been iodinated salts or acids, whose major disadvantage has been high osmolality. For some years now, a new generation of contrast media has been available. These contrast media exhibit a far lower osmolality due to the lack of ionicity. As a result of the volume load of the left ventricle by levocardiography, there is usually an increase of the left ventricular end-diastolic pressure in the pathologic range over 12 mmHg. We were able to prove this in 120 consecutive patients whom we examined. The pressure increase with the use of nonionic contrast media was considerably lower. The differentiated observation of various risk groups showed no signs of valid predictive parameters. PMID- 2568804 TI - Physicochemical properties and general pharmacology of the nonionic dimer iotrolan. AB - Iotrolan, a nonionic, hexaiodinated dimer, is an extremely hydrophilic compound (P = 0.005). Due to its larger Stokes' radius compared with monomeric compounds such as metrizamide, the diffusion time through membranes is extended. Iotrolan deforms erythrocytes only minimally. There is practically no binding to plasma proteins. The new contrast agent has been shown to exert a very limited effect on the complement system (in vitro); it does not inhibit lysozyme (a standard enzyme) in concentrations less than 100 mg I/ml. To inhibit activity of the enzyme collagenase, much higher concentrations of iotrolan than of metrizamide or iopamidol are needed and this could offer an advantage when used for diskography preceding diskolysis with collagenase. After a single intravenous injection in rats, iotrolan has an LD50 of 28.3 g I/kg - the best general tolerance known for water-soluble contrast media thus far. The superior tolerance of iotrolan compared with iohexol and iopamidol (p less than or equal to 0.05) in rats is statistically significant. On the basis of preclinical experience, iotrolan is a very promising contrast medium for intrathecal and intravascular use. PMID- 2568803 TI - Influence of an ionic and a nonionic (sodium meglumine diatrizoate and iopromide) x-ray contrast medium of hemorrheology. AB - In two groups of ten patients each, an ionic and a nonionic contrast medium was injected intravenously, for digital subtraction angiography of the supra-aortic branches, to examine the influence of hemorrheologic properties. Afterward, blood samples were obtained through a central venous catheter and evaluated by a new instrument (oscillating capillary rheometer and densitometer). The viscous and elastic components of the complex blood viscosity and the hematocrit decreased after injection of ionic and nonionic contrast media. A statistically significant difference between the patient groups could not be found. The evaluated results describe an improvement of the hemorrheologic properties after contrast media injection. PMID- 2568805 TI - Organ-specific and general tolerance of iotrolan 280 after intravenous administration: phase I study in healthy volunteers. AB - Iotrolan 280, the first water-soluble, nonionic, blood-isotonic, dimeric contrast medium, was administered intravenously to 12 healthy male volunteers. In a Phase I study with an intraindividual design in comparison with placebo, four doses between 0.15 and a maximum of 0.9 g I/kg body weight were administered in accordance with the principle of dose titration. The highest volume administered was 270.6 ml. The injection rate was 10 ml/min. The observation period was 5 days, with the exception of thyroid parameters (14 days). Iotrolan displayed good general and local tolerance. The typical side effects known from x-ray contrast media either did not occur or were minor after administration of iotrolan. No allergy-like reactions and no effects on hemodynamic parameters were observed. Although one observation is under discussion no effects of iotrolan on impulse generation and propagation in the myocardium could be confirmed. The blood and laboratory and chemical parameters analyzed showed no differences in comparison with placebo. In the treatment group with the highest dose (= 0.9 g I/kg body weight) there was one subject who showed a transient increase of the urinary glucose concentration and one case of a slight transient increase of the serum chloride concentration. No such side effects were seen in the subjects of the treatment groups with lower doses. All renal functions tests were normal. In this study iotrolan showed excellent tolerance after intravenous injection up to 0.9 g I/kg body weight. PMID- 2568806 TI - Hemorrheologic effects of iotrolan after intra-arterial injection in rabbits: comparison with other types of contrast media. AB - Aqueous solutions of the x-ray contrast media (CM) iotrolan and meglumine diatrizoate with and without gelatin added were injected into the rabbit aorta just above the bifuraction in a dose of 0.3 g I/kg and the duration of opacification of the vessel was measured. The difference between the periods recorded for iotrolan and the viscous diatrizoate solution (4 to 6 seconds) and those for the pure diatrizoate solution (3 to 4 seconds) was statistically significant (p less than or equal to 0.05). The differences are directly correlated to the viscosity of the CM solutions. At first the intra-aortal injection of iotrolan causes a somewhat more marked decline in femoral artery blood flow than diatrizoate. The increase in blood flow, which begins 10 seconds after injection is, however, significantly (p less than or equal to 0.05) less after iotrolan than after diatrizoate. This is attributable to the differences in the osmolalities of the two CM. Due to the fact that it is isotonic to blood and because of its viscosity, iotrolan has certain advantages over conventional agents for arteriography that are relevant to clinical practice. PMID- 2568807 TI - Experimental evaluation of radiographic contrast media in perivascular pain receptors in the perfused isolated rabbit ear. AB - The algesic effects of intravascularly injected solutions of sorbitol and ionic and nonionic contrast media (CM) with high and low osmolality were tested in the perfused isolated rabbit ear connected to the body by the great auricular nerve. The threshold for the pain reflex effect was found to be at an osmolality of between 600 and 700 mosm/kg H2O, that is, at about twice the osmolality of blood. This result is consistent with results obtained to date in investigations in the non-anesthetized rat and in diagnostic examinations with CM in humans. The effects of both sorbitol and the ionic CM at a given osmolality were dose dependent only after low doses and not after a high dose. There were no pain reactions after injection of ionic and nonionic CM with a low osmolality. Release of prostacyclin was observed in preliminary tests after injection of ionic substances with high osmolality, but not after injection of CM with low osmolality. PMID- 2568808 TI - Initial experience with a nonionic, dimeric contrast medium (iotrolan) in direct and indirect arteriography: a randomized, intraindividual double-blind study in 60 patients. AB - The new isotonic contrast medium iotrolan has been compared with iopromide in aortofemoral arteriography, selective femoral arteriography, and intravenous digital substraction angiography (DSA). In each case a crossover study design has been chosen with special emphasis on patient comfort. Despite problems in the interpretation of results due to a "hangover" effect in selective peripheral arteriography, it may be concluded: (1) Iotrolan causes significantly less discomfort, such as the feeling of heat and most likely also pain, than iopromide. However, one patient reported slight pain after the injection of iotrolan even when no previous injection of iopromide had been performed; (2) in intravenous DSA contrast appeared 1 to 2 seconds later in the region of interest after the injection of iotrolan compared with iopromide, probably due to the slightly higher viscosity of the former agent; (3) otherwise, no differences between the two agents have been observed. No cardiovascular or other kind of side effects occurred with the exception of one slight allergy-like reaction in 60 patients. PMID- 2568809 TI - Iodine-induced hyperthyroidism after contrast media: animal experimental and clinical studies. AB - The healthy thyroid can adapt to an iodine excess in many different ways. The autoregulation mechanisms may, however, fail in a diseased thyroid and severe hyperthyroidism may result. There must be autonomous portions of the thyroid functionally present to cause an iodine-induced hyperthyroidism. Retrospective and prospective studies, as well as animal experiments, were performed to establish further these pathophysiologic mechanisms. It was demonstrated that 15% of all cases of hyperthyroidism can be attributed to the use of iodine-containing contrast media. Furthermore, all contrast media exhibit a deiodinization during their intracorporeal retention time. The most frequent thyroid disorders, usually of a temporary nature, occur after choledochal contrast media. To some extent, these pathophysiologic processes can be reproduced in the animal model. Beyond this, the animal experimental studies show that it is possible to determine functionally autonomous portions of the thyroid by means of a dynamic thyroid scintigraph. PMID- 2568811 TI - Arterial bolus curve after intravenous injection of ionic and nonionic contrast media. AB - Studies of the intra-arterial bolus curve after intravenous contrast injection with dynamic computed tomography show that a higher bolus maximum can be obtained with a central injection than with a peripheral injection, other injection parameters remaining unchanged. In a singular venous injection ionic and nonionic contrast media show no different effects on the bolus curve. Due to the unfavorable effects of ionic contrast media with high osmolality, for the purposes of intravenous digital substraction angiography nonionic contrast media must be recommended. The subsequent injection of sodium chloride results in a higher bolus maximum. PMID- 2568810 TI - Tolerance of iopromide in angiography: a multicenter study. AB - This prospective multicenter study analyzes the image quality and incidence of undesirable side effects in 2543 film and 1133 intravenous digital subtraction angiographic (DSA) examinations with iopromide. It can be shown that the frequency of undesired side effects in both methods is twice as high in patients at risk as in patients without special risk factors. The most frequent side effects are gastrointestinal and, in cerebral angiography, of a central nervous system nature. The image quality for 99% of the film angiographies was good or better. It could be demonstrated for the DSA that a higher iodine concentration of 370 mg I/ml tends to result in a reduction of the image quality with an increased rate of side effects. PMID- 2568812 TI - Controlled, double-blind study of iosimide, a new, nonionic, monomeric contrast medium, and ioxaglate in cerebral angiography. AB - Iosimide is a new synthetic, nonionic, monomeric contrast medium in which a new substance, triiodine-trimesine acid, is used. It has definite advantages compared with the basic substances used in the past, diaminobenzoic acid and monoaminoisophthalic acid. Due to the high stability, the possibility of the formation of toxicologically questionable, free nuclei amino compounds disappears. Only the slightest probability of a cross-reaction with present antibodies exists, because of the low frequency in the environment. On the basis of the hydrophilic structure of the side chains, there is only a slight chemotoxicity. In a controlled, double-blind study on 100 randomized patients iosimide was tested in cerebral angiography against ioxaglate. Ioxaglate is an ionic contrast medium, popular because of its low osmolality. In this study iosimide exhibited no differences in comparison with ioxaglate with respect to the contrast, the neurological status or the liver or renal tolerance. In examining cardiovascular tolerance there is only a slight tendency toward liver changes with iosimide. Examination of the general tolerance, however, shows a statistically significant lower incidence of sensation of heat and pain with iosimide than with ioxaglate. PMID- 2568813 TI - Coronary arteriography with iotrolan 280, a blood-isotonic, nonionic, dimeric contrast medium: preliminary clinical results. AB - The effects of coronary arteriography with ionic, monomeric diatrizoate (iodine content, 370 mg/ml; osmolality, 2.10 Osm/kg) and nonionic, dimeric iotrolan (iodine content, 280 mg/ml; osmolality, 0.27 Osm/kg) were intraindividually compared in five patients with coronary heart disease. According to an open protocol, both contrast media were injected into the left (LCA) and right coronary arteries (RCA), 8 ml and 5 ml, respectively. Before, during, and 60 seconds after each injection, electrocardiograms (ECG) were recorded and heart rate and aortic pressure were measured. Whereas diatrizoate markedly decreased heart rate (LCA, -36%; RCA, -20%) and aortic pressure (LCA, -26%; RCA, -16%), iotrolan administration kept heart rate virtually unchanged (+/- 1%) and only slightly increased aortic pressure (LCA, 5%, RCA, 8%). After iotrolan injection ECG changes (axis shift of QRS and T waves) were still demonstrable, yet the effects of diatrizoate proved to be more significant (prolongation of QRS and QT intervals as well as shifts of QRS and T axis). The opacification of the coronary arteries was always more pronounced after diatrizoate due to its higher iodine content, but the contrast produced by iotrolan usually was satisfactory. No side effects were observed during the study. Thus, this isosmotic contrast medium prevents bradycardia and hypotension during coronary arteriography. The lower iodine content, however, leads to poorer contrast compared with conventional contrast media. PMID- 2568814 TI - Experimental urography in dogs: diagnostic quality and pharmacokinetic behavior of iotrolan in comparison to nonionic and ionic, monomeric contrast media. AB - A dose of 0.3 g I/kg iotrolan yields much better opacification of the renal parenchyma in dogs (n = 6, intraindividual comparison) than sodium diatrizoate or iopromide, and imaging of the urinary tract is practically equally as good. The plasma iodine levels of all three agents 2 minutes after the bolus injections are 3 mg/ml and the half-lives in the blood are 8 to 10 minutes. The injection of iotrolan causes the hematocrit value to drop by maximum 3.8% 10 minutes after the injection, and corresponding figure after iopromide is 10.8% and that for sodium diatrizoate at 4 minutes is 18.9%. In a further study in dogs (n = 4, intraindividual comparison; dose 0.6 g I/kg) it was found that in the period 10 to 240 minutes after a bolus injection of iotrolan or iopamidol the iodine concentration in the urine was 1.5 to 2 times higher after iotrolan than after iopamidol. Iopamidol causes much higher urinary flow than iotrolan, especially in the first 20 minutes after injection. The properties of iotrolan described here appear to indicate that this novel nonionic, dimeric contrast medium has certain advantages over the monomeric, ionic and nonionic agents for urographic indications. PMID- 2568815 TI - Renal excretion and computed tomography enhancement of iotrolan and iopamidol in dogs. AB - A comparison of the renal excretion and computed tomography (CT) enhancement of nonionic, monomer and dimer contrast media (CM) (iopamidol and iotrolan) was planned to determine if the latter provides additional and prolonged enhancement in dogs. The excretion studies show almost identical performance of both CM with respect to plasma concentration and renal excretion. However, the lower osmolality of iotrolan permits greater renal concentration in the medulla, papilla, and urine. The potential for this further enhancement of the kidney in CT or urography is marginal. PMID- 2568816 TI - Nephrotoxicity of high and low osmolar contrast media: case control studies following digital subtraction angiography in potential risk patients. AB - The urinary excretion of kidney-specific marker proteins before and 120 hours after intravenous injection of either high- or low-osmolar contrast media (CM; diatrizoate, iopamidol 370) was monitored in patients after digital vascular imaging. Inclusion criteria for the randomized clinical study in a total of 40 patients (15 women, 25 men; mean age, 64.5 years) were at least 50 years of age or diabetes mellitus with normal creatinine concentration in serum. Compared with the control period, the elimination of tubular indicator enzymes alanine aminopeptidase, gamma-glutamyltranspeptidase, alkaline phosphatase, as well as of glomerular localized angiotensinase A was significantly higher in all patients after injection of the CM. The most significant differences were observed after 48 hours. In contrast, lysosomal N-acetyl-beta-D-glucosaminidase activity in urine specimens reacted less clearly and appears to be a less sensitive parameter in assessing CM nephrotoxicity. Elimination of brush border as well as of glomerular marker proteins was significantly lower after intravenous injection of low-osmolar CM iopamidol 370 (832 mOsm/kg) than after meglumine diatrizoate 76 (2100 mOsm/kg). In all 40 patients a significant decrease in creatinine clearance was observed; however, patients receiving diatrizoate had a significant decrease in creatinine clearance (period 0 versus 24 to 48 hours after CM), whereas patients after administration of iopamidol had not. No difference was found between creatinine clearance after 48 hours of CM injection within both groups of CM. Due to noninvasive parameters of kidney damage nonionic, low-osmolar CM are less nephrotoxic in potential risk patients, and should be preferred to conventional CM. PMID- 2568817 TI - Renal and hepatic tolerance of nonionic and ionic contrast media in intravenous digital subtraction angiography. AB - The liver and kidney tolerance of iopromide 370 in comparison to that of sodium meglumine diatrizoate 370 or iopamidol 370 in doses of 2 ml/kg body weight was examined in two controlled double-blind studies with intravenous digital subtraction angiography on the basis of enzyme assays in serum and urine. In patients with normal kidney function no changes were observed in the levels of the liver enzymes GPT, GOT, and gamma glutamyl transpeptidase (GGT) serum up to 72 hours after injection of iopromide or sodium meglumine diatrizoate. Among the kidney-specific enzymes, the excretion of GGT in urine increased after injection of iopromide and iopamidol. The maximum increase of GGT excretion was, however, statistically significantly lower in the group treated with iopromide than in the iopamidol group. Within 72 hours, the activities had been returned to the initial values in both groups. PMID- 2568818 TI - Induction of ketone body enzymes in glial cells. AB - Ketone bodies serve a dual function in developing brain. They are important sources of energy for metabolism and serve as precursors for lipid synthesis. Astrocytes have two to three times higher activity than oligodendroglia for one of the enzymes involved in ketone body metabolism, 3-ketoacid-CoA transferase. Both glial cell types have similar levels of activity for beta-hydroxybutyrate dehydrogenase. Glucocorticoids and dibutytyl cAMP produce a significant stimulation of activity of both enzymes in astrocytes and oligodendroglia. However, the most striking induction in activity of the two enzymes is in the presence of hydrocortisone and sodium butyrate. There is a three- to eightfold stimulation with these effectors in both astrocytes and oligodendroglia. Thus, in brain the expression of ketone body enzyme activities is finely regulated by hormones and by agents that increase cAMP levels. PMID- 2568819 TI - Examination of the role of thiolimidate formation in the cleavage of acetoacetyl CoA catalyzed by thiolase I from porcine heart. AB - The potential contribution of thiolimidate formation to the increased kinetic acidity of the alpha-proton of acetyl-CoA in the carbon-carbon bond forming reaction catalyzed by 3-ketoacyl-CoA thiolase (thiolase I) from porcine heart was assessed by chemical modification and isotope exchange experiments. Thiolase is only partially inactivated after the chemical modification of lysine residues by reductive methylation, pyridoxal phosphate, or o-phthaldehyde (specific for vicinal lysine and cysteine). The thiolase-catalyzed formation of acetyl-CoA from acetoacetyl-CoA and CoASH in 18OH2 is not accompanied by the appearance of 18O in the acetyl-CoA product. These experiments effectively rule out participation of thiolimidate formation in the thiolase reaction. Other mechanisms must be employed to facilitate the abstraction of the alpha-proton of acetyl-CoA by thiolase I. PMID- 2568820 TI - [A case of peritoneal malignant mesothelioma with disappearance of ascites result of intraperitoneal instillation of mitomycin C and oral administration of UFT]. AB - We reported a 65-year-old man with peritoneal malignant mesothelioma which was discovered by hydrocele of left undescended testis. Initially, intraperitoneal instillation of CDDP 100 mg/body accompanied with aspiration of ascites was not effective. Secondarily, instillation of MMC 10 mg/body every other week was performed five times, and ascites completely disappeared. After that oral administration of UFT 300 mg/day, which was added to instillation of MMC 10 mg/body every four week, has been performed. Cytology of ascites changed to negative, and the clinical remission has continued ever since. PMID- 2568821 TI - [The first Soviet-American Symposium on Antibiotics and Chemotherapy]. PMID- 2568822 TI - Impaired DNA synthesis by early pubertal cryptorchid testis in vitro. AB - To study the spermatogenic potential of the early pubertal cryptorchid testis, testicular DNA synthesis was examined in vitro. DNA synthesis by early pubertal cryptorchid testis was inhibited and lost its normal temperature sensitivity. The spermatogenic potential of the cryptorchid testis may be impaired as early as puberty. PMID- 2568823 TI - Diagnosis of peripheral androgen insensitivity in a male infant excretion analysis. AB - The hypothesis of peripheral androgen insensitivity (AIS) was examined in a boy with congenital growth hormone deficiency associated with micropenis and cryptorchidism by steroid excretion analyses compared with dihydrotestosterone (DHT) receptor analyses of foreskin biopsy homogenate. Urinary T metabolite 3 alpha,17 beta-dihydroxy-5 beta-androstane (3 beta-diol) was below the limit of detection (capillary gas chromatography) at age 1 year, but on several occasions (n = 7) normal basal values were found at age 3 years (patient: 78.9 +/- 25.4 micrograms/24 h, M +/- SD; controls, n = 15: 100.5 +/- 50.2). Normal basal- and hCG-induced excretion was noted for 3 alpha,17 beta-dihydroxy-5 alpha-androstane (3 alpha-diol) at age 1 and 3 years, respectively. Additionally, basal 3 alpha diol excretion (n = 7) at age 3 years was 66.7 +/- 21.1 micrograms/24 h, M +/- SD; controls (n = 15) 75.8 +/- 50.4. Analysis of urinary androgens might be an alternative, noninvasive procedure for the diagnosis of peripheral AIS. PMID- 2568824 TI - The positive-negative distinction in drug-free schizophrenic patients. Stability, response to neuroleptics, and prognostic significance. AB - Fundamental questions about the validity and significance of positive and negative syndromes in schizophrenia were addressed by a prospective, double-blind longitudinal study that involved a drug-free placebo baseline, three to four months of neuroleptic treatment, and a three-year poststudy follow-up. From pooled data on 62 schizophrenics, the following findings were observed: (1) a high stability of both syndromes during drug-free conditions; (2) significant correlations of syndrome ratings between the placebo baseline and final neuroleptic week; (3) significant neuroleptic-related improvement in both positive and negative syndromes, with a marginally greater reduction of positive features; (4) independence of the two syndromes during the drug-free baseline but not under neuroleptic conditions; (5) greater symptomatic improvement but more residual disorder portended by both positive and negative syndromes in the drug free baseline; and (6) poorer functional reconstitution and earlier relapse predicted by a positive syndrome alone. These data supported the validity of the positive-negative distinction in schizophrenia but challenged basic assumptions about its import. PMID- 2568825 TI - Tardive dyskinesia in Italy: preliminary findings. PMID- 2568826 TI - Saccadic distractibility in schizophrenic patients with tardive dyskinesia. PMID- 2568829 TI - Assistants at cataract surgery. PMID- 2568828 TI - When does the failure to find a difference mean that there is none? AB - When a published clinical report concludes that there is no difference between two or more groups of patients, the reader must discern between two alternative possibilities. Either there is truly no difference in outcome between the groups or the study failed to detect an actual difference. The application of sample size and power calculations to outcome variables in a clinical study can provide essential insight into resolving this ambiguity. While one can never prove that there is no difference between groups of patients, these techniques can estimate the likelihood that a particular study will detect or fail to detect a difference of specified size. PMID- 2568827 TI - Characteristics of passive immunity against hantavirus infection in rats. AB - The protective effects of passively administered antibodies against hantavirus infection were studied in newborn rats. Death as well as infection were completely prevented from intraperitoneal challenge of strain SR-11 (SR) (2 x 10(3) FFU, 10(2.1) LD50), in newborn rats which received 0.1 ml of anti-SR rat serum (neutralizing antibody titer, 1:640) 4 hr before the virus challenge. In these rats, no virus was detected in the peritoneal macrophages, lung, kidney, and brain. The immune serum infusion before the virus challenge also conferred protection to rats against an intramuscular or subcutaneous challenge of strain SR, but did not protect the rats against intracerebral challenge of the virus. In the rats which received the immune serum after the challenge, infection was not prevented, although some of the animals were protected from the death. Virus titers in the lung, kidney, and brain of the rats were reduced by the transfer of the immune serum even as late as 72 hr after the challenge. Cross-protection in the rats which received the immune serum was strong between strains SR and KI-262 within the same serotype, but very weak between strains SR and Hantaan 76-118 of different serotypes. PMID- 2568830 TI - Prohibition on charges for services of assistant cataract surgeons upheld by Federal Court of Appeals. PMID- 2568831 TI - Role of alpha-adrenergic stimuli in the control of rat renal ammoniagenesis. AB - The effects of phenylephrine on renal ammoniagenesis and the involvement of Ca2+ in phenylephrine action were investigated in isolated proximal fragments of rat kidney tubules. Phenylephrine stimulated renal ammoniagenesis from 1 and 2 mM glutamine whereas no significant changes took place at a higher concentration of glutamine (20 mM). Stimulation of ammonia synthesis by phenylephrine was found to be linear with time and dose-dependent between 10(-9) and 10(-4) M. Phenylephrine stimulated ammoniagenesis was blocked by phentolamine (10 microM) but not by propranolol (10 microM) confirming that the effect is mediated by alpha adrenergic stimuli. No stimulatory effect of phenylephrine was observed in Ca2+ depleted proximal tubule fragments, suggesting that Ca2+ is required in this adrenergic response. PMID- 2568832 TI - P-glycoprotein gene (MDR1) cDNA from human adrenal: normal P-glycoprotein carries Gly185 with an altered pattern of multidrug resistance. AB - We isolated a full-length MDR1 cDNA from human adrenal where P-glycoprotein is expressed at high level. The deduced amino acid sequence shows two amino acid differences from the sequence of P-glycoprotein obtained from colchicine-selected multidrug resistant cultured cells. The amino acid substitution Gly----Val at codon 185 in P-glycoprotein from colchicine resistant cells occurred during selection of cells in colchicine. As previously reported, cells transfected with the MDR1 cDNA carrying Val185 acquire increased resistance to colchicine compared to other drugs. The other amino acid substitution Ser----Ala at codon 893 probably reflects genetic polymorphism. The MDR1 gene, the major member of the P glycoprotein gene family expressed in human adrenal, is sufficient to confer multidrug-resistance on culture cells. PMID- 2568834 TI - [A modification of neurotoxin RTX-III from Radianthus macrodactylus actinin with acetic anhydride]. AB - Upon modification of neurotoxin RTX-III at amino groups with [3H]acetic anhydride, four monoacetyl and four diacetyl derivatives have been obtained. Acetylation of the N-terminal amino group led to 12-fold decrease of toxicity in mice. Monoderivatives of the toxin with either Lys or two out of three C-terminal Lys residues modified showed a 2-fold drop in toxicity as compared with the native RTX-III. Diacetylation caused a 30 to 35-fold decrease in toxicity, the N terminal amino group being modified in all the derivatives. As assessed by circular dichroism method, the modification of amino groups, except for N terminal one, affected secondary structure of the toxin. The data suggest the N terminal amino group to be essential for toxicity of RTX-III. PMID- 2568833 TI - Identification of P-glycoprotein in renal brush border membranes. AB - A monoclonal antibody (C219) that recognizes the P-glycoprotein (Mr = 170,000) in plasma membranes of multidrug-resistant Chinese hamster ovary (CHO) cell lines was used to assay renal brush border membrane (BBM) and basolateral membrane (BLM) fractions for the presence of a cross-reactive polypeptide. The C219 antibody bound to a 155,000 dalton protein in immunoblots of rat BBM but not BLM proteins resolved by sodium dodecyl sulfate gel electrophoresis. The corresponding human kidney BBM and dog kidney BBM proteins had molecular weights of 170,000 and 160,000 respectively. The glycoprotein nature of the renal protein was shown by its sensitivity to N-glycanase treatment which reduced the apparent molecular weight of the dog protein to 120,000. In addition, dog P-glycoprotein could be bound to and eluted from immobilized wheat germ agglutinin. The molecular weight, antibody crossreactivity, glycosidase sensitivity and lectin binding show that this protein is a normal kidney analogue of the P-glycoprotein induced in multidrug resistant cell lines. PMID- 2568835 TI - Characterization of the class I HLA 9.2-kb PVU II restriction fragment length polymorphism. Linkage to HLA-A and lack of disease association. AB - The strongest reported association between a class I HLA allele and disease is that of HLA-B27 with ankylosing spondylitis (AS). However, it has not been shown whether B27 is the gene that predisposes to the development of AS or if it is merely linked with the disease-susceptibility locus. Furthermore, if B27 itself is the disease-susceptibility gene, there may be epistatic loci that also contribute to the development of AS or modify its clinical manifestation. A class I HLA 9.2-kb Pvu II restriction fragment was recently identified, which, when present in a B27-positive individual, further increased the relative risk for developing AS (from 119 to 297). This study was therefore designed to confirm the association between AS and this restriction fragment length polymorphism (RFLP) and to map the location of this fragment in the genome. The data presented here suggest that the class I HLA 9.2-kb Pvu II RFLP represents a Pvu II polymorphism at the 5' end of the HLA-A locus that is tightly linked with both HLA-A3 and A9 alleles. However, there is no association between this RFLP and AS in a population of patients living in Baltimore. PMID- 2568836 TI - Cardiohemodynamic effects of bopindolol in normotensive conscious dogs. A comparative study with pindolol. AB - Cardiohemodynamic effects and the beta-blocking action of a new beta-adrenoceptor blocking agent, bopindolol (Sandonorm) were compared with those of pindolol in conscious unrestrained dogs. The intravenous injection of pindolol (3-100 micrograms/kg) increased the heart rate and decreased the total peripheral resistance dose-dependently and markedly, while the same doses of bopindolol had no effect on the heart rate and decreased the total peripheral resistance only at the maximum dose (100 micrograms/kg). These changes were antagonized by propranolol (3 mg/kg). Isoprenaline (isoproterenol, 0.1 microgram/kg)-induced tachycardia was inhibited dose-dependently by both beta-blockers to similar degrees. While the decrease in the total peripheral resistance by isoprenaline was inhibited by pindolol dose-dependently, the inhibition by lower doses of bopindolol was not so marked; the inhibition became greater as the doses of bopindolol were increased. These results indicate that bopindolol is a beta 1 selective beta-blocker with an affinity to this subtype of the beta-adrenoceptor comparable to that of pindolol. Unlike pindolol, it exerted no partial agonist activity at the beta-adrenoceptor. Instead it produced the partial agonist activity at the beta 2-adrenoceptor at the highest dose tested. PMID- 2568837 TI - Toxicological evaluations of the benzodiazepine doxefazepam. AB - 1-(2-Hydroxyethyl)-3-hydroxyl-7-chloro-1,3-dihydro-5-(O-fluorophenyl)-2H - 1,4 benzodiazepin-2-one (doxefazepam, SAS 643, Doxans) was investigated in a series of toxicological studies. Oral LD50 values were greater than 2000 mg/kg in mice, rats and dogs, while endoperitoneal LD50 values were 746 and 544 mg/kg in the mice and rats, respectively, and greater than 1000 mg/kg in the dogs. Subacute and chronic studies in rats and dogs evidenced a transient ataxia after administration of the test compound, which was dose-dependent in the subacute experiment, and occurred only at the highest dose in the chronic studies. No pathological findings were registered at necropsy or in microscopic observations, except an increase of liver weight at the highest dosage in the chronic study in the rat. Doxefazepam did not exert any teratogenic effects in rats and rabbits. Moreover in rats it did not alter the reproductive performance. The mutagenic studies did not reveal any mutagenic potential. In the cancerogenicity study in rats doxefazepam did not show positive carcinogenic potential. PMID- 2568838 TI - [Beta receptor block versus calcium antagonism. A comparative study of bopindolol and nifedipine with special regard to quality of life]. AB - 203 patients with a diastolic blood pressure higher than 100 mmHg were included in a randomized, double-blind trial to compare the antihypertensive efficacy of different daily dosages of bopindolol (Wandonorm) (0.05 mg, 0.5 mg, 1 mg, 2 mg) and nifedipine (2 x 20 mg). After 4 weeks of therapy blood pressure normalization could be achieved in 23.7% (0.05 mg), 32.5% (0.5 mg), 67.5% (1 mg), 64.1% (2 mg) and 59.0% (nifedipine) of the patients, respectively. In a subgroup of 159 patients the study was continued with an 8-week dose titration and a 16-week observation. At the end of the study blood pressure normalization was achieved in 91% and 94% of the patients treated with nifedipine and bopindolol, respectively. Most patients of the bopindolol-group needed 1 mg once daily as compared to those on the nifedipine who required 20 mg b.i.d. Because of intolerable side effects therapy was discontinued in 3 out of 162 patients on bopindolol and in 3 out of 41 patients on nifedipine. As compared to nifedipine the tolerance of bopindolol was judged significantly superior because tiredness and dizziness (32% vs 9%) and leg edema (20% vs. 6%) were recorded much more frequently in nifedipine treated patients. In 135 elderly patients "quality of life" was assessed upon by use of the "Nuremberg-Alters-Selbstbeurteilungs-Skala" (NAS), which refers to social contacts, mental and physical performance, sleep disturbances and general well being.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568839 TI - Restriction fragment length polymorphisms in the apo B gene in relation to coronary artery disease. AB - We have determined the frequencies of the alleles at the EcoRI (E), XbaI (X) and PvuII (P) polymorphic restriction sites in the apo B gene in 124 white men with coronary artery disease (CAD) and in 146 white men free from CAD. The frequencies of the E- (restriction site absent) and X- alleles were both significantly higher in normocholesterolaemic men with CAD than in those without CAD, but the frequency of the P+ allele (restriction site present) was similar in the 2 groups. The frequency of the E- allele was significantly higher in CAD men with hypertriglyceridaemia than in normal men without hypertriglyceridaemia. In the normocholesterolaemic men without CAD, the mean serum cholesterol concentration was higher in those with genotype X++ than in those with genotype X--. Mean serum LDL-apo B and LDL-cholesterol concentrations did not differ significantly between men with different XbaI or EcoRI genotypes. Serum apo A-I levels differed significantly between normal men with different XbaI genotypes. Serum HDL cholesterol levels differed significantly between CAD men with different XbaI genotypes. These results suggest that in white men the E- and X- alleles are in linkage disequilibrium with a nearby allele that is causally related to CAD. It is also possible that the amino acid substitution at position 4154 in apo B, brought about by the nucleotide change responsible for the EcoRI polymorphism, has a direct effect on the atherogenicity of LDL. PMID- 2568840 TI - [Prolonged action of vecuronium in neurofibromatosis (von Recklinghausen's disease)]. AB - It is reported on an abnormally prolonged action of vecuronium in a female patient with known neurofibromatosis (Morbus von Recklinghausen). In this female patient an increased sensitivity to vecuronium (0.06 mg/kg) was observed during anaesthesia for Caesarean section. A 100% block lasted 50 minutes and the neuromuscular recovery from 75% block to 25% block amounted to 25 minutes. This prolonged neuromuscular block can also be due to overdosing, relative hypovolaemia during arterial hypertension and the previous application of suxamethonium. PMID- 2568841 TI - Vaccination against experimental autoimmune encephalomyelitis using a subencephalitogenic dose of autoimmune effector T cells. (2). Induction of a protective anti-idiotypic response. AB - We previously reported that a subencephalitogenic dose (10(4) of activated anti BP Z1a T cells rendered Lewis rats significantly resistant to EAE induced either actively or adoptively. This resistance was specific to EAE and persisted for over 4 months. The experiments reported in this paper were done to investigate the mechanisms of this resistance. We found that the state of vaccination was marked by a decrease in the in vitro proliferation and in vivo DTH responses to BP. Resistance could be transferred to recipient rats with the thymus or spleen cells of donor vaccinated rats. Vaccination led to the appearance of proliferative and DTH responses that were specifically directed to the Z1a T cells. The kinetics and compartmentalization of this anti-idiotypic responsiveness was studied by vaccinating rats in the hind footpads and monitoring the proliferative reactivity of the draining popliteal lymph node (PLN) and distal cervical lymph node (CLN) cells at various times. We found that the anti-idiotypic reactivity was confined to the PLN on days 5-6 and thereafter became systemic. Excision of the PLN on day 6, but not on days 3 or 11, robbed the rats of their acquired resistance to EAE. In contrast, the PLN cells of the vaccinated rats transferred resistance to naive donors. Thus, the lymphoid population containing cell-mediated anti-idiotypic responsiveness served as a vehicle of resistance. These results suggest that anti-idiotypic T-cell immunity to autoimmune effector T cells is involved in the resistance to EAE induced by T cell vaccination. PMID- 2568842 TI - Functional heterogeneity of CD4+ T cells in leishmaniasis. AB - In this article F.Y. Liew summarizes the evidence for the functional heterogeneity of CD4+ T cells from studies of immune regulation in the parasitic disease, leishmaniasis. Recent findings suggest that the heterogeneity of CD4+ T cells may extend beyond the current Th1 and Th2 classification and that these T cell subsets manifest some of their functions via the lymphokines they secrete, and that the balance of these determines the outcome of the infection. PMID- 2568844 TI - Advanced immunodeficiency as a risk factor for heterosexual transmission of HIV. AB - To define the rate of HIV infection and associated risk factors among heterosexual partners of HIV-infected individuals, 80 partners and 77 index cases were examined for clinical, behavioural and virological variables. The study population included 61 male and 16 female index cases, of whom 40 were European and 37 African. Multiple sexual partners in central Africa was the risk factor for HIV infection in 83%. The overall infection rate among the contact cases was 45%, 53% among female partners and 13% among male partners (P less than 0.001). Variables significantly associated with HIV infection in the partners included advanced clinical stage (AIDS-related complex or AIDS), a low concentration of T4 lymphocytes and African nationality of the index case, young age of the partner, and more than 50 instances of sexual contact with the index cases. In a logistic regression analysis, a low concentration of T4 lymphocytes, clinical stage ARC/AIDS and African nationality of the index case but not number of sexual contacts, remained as independent risk factors for HIV transmission. Our data indicate that there is a significant biological heterogeneity in sexual transmission of HIV and that advanced clinical stage and severe T cell depletion of the index case are major determinants of infectivity. PMID- 2568843 TI - Pro-opiomelanocortin gene expression in human phaeochromocytomas. AB - Phaeochromocytoma is an occasional cause of the ectopic ACTH syndrome. The mechanisms of proopiomelanocortin (POMC) gene expression were analysed in 11 human tumours not associated with Cushing's syndrome, by detecting and characterizing the POMC mRNA. A DNA probe corresponding to most of the protein coding region of the third exon was used in Northern blot studies of total and poly(A)+ RNA. All tumours contained a short (800 bases) mRNA species different from the 1200 base mRNA species of the human pituitary. This short mRNA was also present in the normal adrenal, where S1 mapping showed that it resulted from transcription initiation within the third exon. However, in two tumours, equivalent amounts of the 1200 base mRNA were also present, and in one of them a third POMC mRNA of approximately 1450 bases was detected. These data show that POMC gene expression occurs in all phaeochromocytomas. It is suggested that excess production of the 1200 bases (or the larger, 1450 base) mRNA in some tumours may be responsible for the rare occurrence of the ectopic ACTH syndrome. PMID- 2568845 TI - Genetic analysis of CR1 (the C3b complement receptor, CD35) expression on erythrocytes of HIV-infected individuals. AB - The number of C3b receptor (CR1) molecules on erythrocytes is genetically determined by two codominant autosomal alleles. The genetic polymorphism of CR1 expression correlates with a Hind III restriction fragment length polymorphism (RFLP) of the CR1 gene. The relative frequency of individuals homozygous for the allele coding for low CR1 numbers is approximately 5% of the normal population. CR1 numbers/erythrocytes are significantly decreased in symptomatic HIV-infected individuals. Decreased CR1 expression correlates with the severity of disease. The present study investigated the CR1 genomic Hind III RFLP-related polymorphism in 79 HIV-infected individuals and 84 healthy subjects. Allele frequencies were found to be similar in both populations. Thus, there is no susceptibility nor resistance to HIV-infected linked to the CR1 gene. Defective expression of CR1 in HIV-infected patients is acquired through central and/or peripheral mechanisms. PMID- 2568846 TI - Left ventricular function in hypertension. Relevance to the selection of antihypertensive therapy. AB - With the increasingly widespread recognition of the adverse side effects of traditional antihypertensive medications, alternative approaches to the treatment of uncomplicated hypertensive patients have been proposed. The use of vasodilator agents, calcium-channel blockers, angiotensin-converting enzyme (ACE) inhibitors, and alpha-adrenergic receptor blockers results in blood pressure reduction without adverse metabolic consequences. Furthermore, vasodilatory agents reverse the principal physiological abnormality of hypertensive patients, that is, they reverse the increased systemic vascular resistance that characterizes these patients and, in so doing, may alleviate abnormalities of systolic and diastolic left ventricular performance. In particular, a comparison of the effects of nifedipine and propranolol on left ventricular (LV) function in patients with moderately severe hypertension revealed significant differences in their effects on cardiac function. Although both drugs reduced blood pressure to an identical degree, nifedipine therapy was associated with a decrease in systemic vascular resistance, an increase in cardiac output, and improved parameters of left ventricular contractile and diastolic function as measured by quantitative radionuclide ventriculography. Vasodilator therapy should be considered for the initial treatment of patients with uncomplicated essential hypertension. PMID- 2568848 TI - Use of atracurium or vecuronium to prolong the action of tubocurarine. AB - Forty patients received tubocurarine in a dose greater than ED90. When neuromuscular function had recovered to amplitude of the first contraction of the train-of-four equals 10% of control, a small increment of atracurium or vecuronium was administered, repeating the same increment at each subsequent recovery to 10%. The intensity and duration of the neuromuscular block following the first increment was always greater than that of subsequent increments. The duration and intensity of the block was progressively reduced with subsequent increments until the responses to further increments were unchanged. These final means at steady state were: group 1 (atracurium 1.1 mg) 6.4 (0.3) min; group 2 (atracurium 2.0 mg) 8.2 (0.9) min; group 3 (vecuronium 0.25 mg) 5.8 (0.4) min; group 4 (vecuronium 0.5 mg) 13.2 (0.4) min. PMID- 2568847 TI - Localization of acyl-coenzyme A: cholesterol acyltransferase in villus and crypt cells of chick intestine. AB - Endogenous cholesterol esterification by acyl-CoA:cholesterol acyltransferase (EC 2.3.1.26) was studied in isolated enterocytes obtained from chick duodenal, jejunal, and ileal villi and crypts, using [14C]oleoyl-CoA as substrate. The maximal specific activity in each cell fraction was found in chick jejunum, followed by duodenum and ileum. Jejunal upper and mid villi showed higher specific activities than lower villi and crypts. Epithelial cells isolated from chick intestine also incorporated oleoyl-CoA into different lipids using the endogenous substrates. Upper and mid villus cells showed the maximal incorporation of oleoyl-CoA into triglycerides in duodenum and jejunum. Levels of oleoyl-CoA incorporation into phospholipids were higher than those found in the synthesis of triglycerides or cholesterol esters, whatever may be the cell fraction considered. Upper villus cells also showed the highest specific activity in the incorporation of oleoyl-CoA into phospholipids. The acyl-CoA hydrolase specific activity was practically similar in all the cell fractions obtained from chick duodenum, jejunum, and ileum. PMID- 2568849 TI - Accelerated recovery from combined atracurium-vecuronium neuromuscular block. AB - Patients given combinations of non-depolarizing neuromuscular blocking drugs have been reported to recover from neuromuscular block more rapidly than patients given a single drug. This study was designed to assess if this phenomenon occurred with the combination of atracurium and vecuronium. During nitrous oxide fentanyl anaesthesia, 30 adult patients were allocated randomly to receive atracurium 0.5 mg kg-1, vecuronium 0.1 mg kg-1, or a combination of atracurium 0.125 mg kg-1 + vecuronium 0.025 mg kg-1. All patients had 100% neuromuscular block, and times to block onset did not differ significantly between the three groups. Recovery to 10, 25, 50 and 90% of control twitch height was significantly faster in the group receiving the combination of drugs. PMID- 2568850 TI - Phenothiazine-binding and attachment sites of CAPP1-calmodulin. AB - In the presence of Ca2+ norchlorpromazine isothiocyanate forms a monocovalent complex with calmodulin: CAPP1-calmodulin (Newton et al, 1983). Trypsin digestion of [3H]CAPP1-calmodulin yields as the major radioactive peptide N epsilon-CAPP Lys-Met-Lys, corresponding to residues 75-77 of calmodulin. Stoichiometric amounts of all other expected tryptic peptides are also found, indicating that norchlorpromazine isothiocyanate selectively acylates Lys 75. A second molecule of CAPP-NCS can react, albeit slowly, with calmodulin to form CAPP2-calmodulin. Fragments 38-74 and 127-148 are completely missing from the trypsin digests of CAPP2-calmodulin without deliberate exposure to UV irradiation. Possibly the lengthy preparation of CAPP2-calmodulin favors photolysis, caused by room lights, of the putative CAPP-binding domains located in these two peptides. Lys 148, the sole lysyl residue in fragment 127-148, is a probable site of attachment of the second molecule of CAPP. UV irradiation of CAPP1-calmodulin, followed by digestion with trypsin, results in the selective loss of 50% each of peptides containing residues 38-74 and 127-148, suggesting that these peptides contain the hydrophobic amino acids that form the phenothiazine-binding sites. The loss of peptides encompassing residues 38-74 and 127-148, located in the amino and carboxyl halves of calmodulin, respectively, suggests that the hydrophobic rings of CAPP can bind at either one of the two phenothiazine sites. Computer modeling of CAPP1-calmodulin with the X-ray coordinates of calmodulin (Babu et al., 1986) indicates that CAPP attached to Lys 75 cannot interact with the carboxyl-terminal phenothiazine-binding site.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568851 TI - Aspartate aminotransferase catalyzed oxygen exchange with solvent from oxygen-18 enriched alpha-ketoglutarate: evidence for slow exchange of enzyme-bound water. AB - Partitioning of the ketimine (or ketimine + quinonoid) intermediate(s) in the mitochondrial aspartate aminotransferase reactions was investigated by following the rates of loss of 18O from carbonyl-18O-enriched alpha-ketoglutarate together with the rate of L-glutamate formation. The ratio of these rate constants was found to equal 1 at 10 degrees C, implying that the above intermediate(s) face(s) equal barriers with respect to the forward and reverse reactions. This partition ratio of 1 together with that measured from the alpha-amino acid side of the reaction [Julin, D.A., Wiesinger, H., Toney, M. D., & Kirsch, J.F. (1989) Biochemistry (preceding paper in this issue)] suggests that the rate constant for exchange of alpha-ketoglutarate-derived H2(18)O from the ketimine (or ketimine + quinonoid) form(s) of the enzyme with solvent is comparable with that for kcat. PMID- 2568852 TI - The structure and function of the homeodomain. PMID- 2568853 TI - Transamination and oxidative decarboxylation of L-isoleucine, L-alloisoleucine and related 2-oxo acids in perfused rat hind limb muscle. AB - Metabolism of L-isoleucine, L-alloisoleucine and corresponding 2-oxo acids in rat hind limb muscle was comparatively studied under steady-state perfusion conditions. At 0.5 mM L-[1-14C]isoleucine, apparent transamination and 2-oxo acid decarboxylation rates amounted to about 17 and 4 nmol/min per g of muscle, respectively. With L-allo[1-14C]isoleucine, the corresponding rates were about 5- and 10-fold lower, respectively. After addition of dichloroacetate (1-5 mM), the portion of (S)- and (R)-methyl-2-oxopentanoate undergoing further oxidative decarboxylation within the tissue was similarly increased by over 40%. In perfusions with 0.5 mM (R,S)-3-methyl-2-oxopentanoate and tracer doses of 1-14C labeled (S)- or (R)-enantiomer, the 14CO2 production was comparable (about 0.5 nmol/min per g of muscle). Dichloroacetate caused a several-fold increase in 14CO2 release from either enantiomer, apparent 2-oxo acid transamination rates remaining unaffected. Indications for a racemization of 2-oxo acid were not obtained in the experiments. The results are discussed with respect to the appearance/disappearance of L-alloisoleucine in vivo and to the fact that (R)-3 methyl-2-oxopentanoate, but not L-alloisoleucine, can support growth of rats on a diet deficient in L-isoleucine. PMID- 2568854 TI - Sulfhydryl groups of an extramitochondrial acetyl-CoA hydrolase from rat liver. AB - An extramitochondrial acetyl-CoA hydrolase (EC 3.1.2.1) purified from rat liver was inactivated by heavy metal cations (Hg2+, Cu2+, Cd2+ and Zn2+), which are known to be highly reactive with sulfhydryl groups. Their order of potency for enzyme inactivation was Hg2+ greater than Cu2+ greater than Cd2+ greater than Zn2+. This enzyme was also inactivated by various sulfhydryl-blocking reagents such as p-hydroxymercuribenzoate (PHMB), N-ethylmaleimide (NEM), 5,5'-dithiobis(2 nitrobenzoic acid) (DTNB), and iodoacetate (IAA). DL-Dithiothreitol (DTT) reversed the inactivation of this enzyme by DTNB markedly, and that by PHMB slightly, but did not reverse the inactivations by NEM, DTNB and IAA. Benzoyl-CoA (a substrate-like competitive inhibitor) and ATP (an activator) greatly protected acetyl-CoA hydrolase from inactivation by PHMB, NEM, DTNB and IAA. These results suggest that the essential sulfhydryl groups are on or near the substrate binding site and nucleotide binding site. The enzyme contained about four sulfhydryl groups per mol of monomer, as estimated with DTNB. When the enzyme was denatured by 4 M guanidine-HCl, about seven sulfhydryl groups per mol of monomer reacted with DTNB. Two of the four sulfhydryl groups of the subunit of the native enzyme reacted with DTNB first without any significant inactivation of the enzyme, but its subsequent reaction with the other two sulfhydryl groups seemed to be involved in the inactivation process. PMID- 2568856 TI - A kinetic fluorometric assay of dipeptidyl peptidase IV in viable human blood mononuclear cells. AB - A continuous-rate fluorometric assay of dipeptidyl peptidase IV (DP-IV) in viable human blood mononuclear cells using 7-(L-glycyl-L-prolylamido)-4-methylcoumarin as the substrate is described. The assay method is accurate, rapid, and highly sensitive for measuring the level of cell-surface bound DP-IV activity in suspension of blood mononuclear cells, as well as of other viable cells bearing this enzyme. We believe that the kinetic assay is suitable for studying the regulation of expression and the role of plasma membrane-bound DP-IV on the cellular level. PMID- 2568855 TI - Spin-labeled analogues of ATP, ADP and AMP: substitutes for normal nucleotides in biochemical systems. AB - The different roles and effectiveness of adenosine monophosphate, diphosphate and triphosphate labeled at the 6 position of the purine ring with 2,2,6,6 tetramethylpiperidine-1-oxyl in reactions catalyzed by Escherichia coli glutamine synthetase (GS) have been investigated. Our results show that the spin-labeled ATP (Tempo-ATP) serves as a substrate in the glutamine synthesis reaction and in the adenylation of E. coli glutamine synthetase catalyzed by ATP: glutamine adenylyl transferase (ATase) with essentially the same effectiveness as normal ATP. In another reaction (gamma-glutamyltransferase), Tempo ADP serves as an effector with a Km of 9.4 . 10(-8) M compared to 1.2 . 10(-8) M for the normal ADP, while covalently bonded Tempo-AMP serves as a modifier on the catalytic properties of E. coli glutamine synthetase just as the covalently bonded normal AMP does. The dissociation constants between the labeled nucleotides, Mn2+, Mg2+ and Ca2+ are in the same order of magnitude as the binding constants for those cations and the corresponding normal nucleotides. Our findings indicate that the spin-labeled nucleotides are good substitutes for the normal nucleotides in the biochemical systems studied. PMID- 2568857 TI - In vitro-perifused rat testes secrete beta-endorphin and dynorphin: their effect on testosterone secretion. AB - Leydig cells of many species synthesize and secrete opioid peptides, but the Sertoli and possibly the peritubular cells are the only intratesticular cells having opiate receptors. It is known that Sertoli and peritubular cells can modify the secretion of testosterone from Leydig cells. To test the hypothesis that testicular opioid peptides participate in a Leydig-Sertoli-peritubular Leydig cell feedback loop that can regulate the intratesticular concentration of testosterone, we have developed a method for the in vitro perifusion of rat testicular fragments in which the intratesticular structure and thus the paracrine feedback loop remains intact. Our data show that both immunoreactive (IR)-beta-endorphin and IR-dynorphin were present in the testicular perifusion effluent; gel chromatography of pooled perifusion effluent show that the bulk of the secreted IR-beta-endorphin had the apparent mol. wt. of synthetic rat beta endorphin whereas most of the secreted IR-dynorphin was composed of smaller than 4000 mol. wt. forms. On the other hand, the bulk of IR-dynorphin present in rat testicular tissue homogenates eluted in two higher mol. wt. peaks. The effect of mu and kappa opioid agonists and naloxone (a universal opioid antagonist) on both basal and gonadotropin-stimulated testosterone secretion from perifused testicular fragments was then examined; no stimulatory or inhibitory effect of the opioid receptor agonists or naloxone was found on basal and gonadotropin stimulated testosterone secretion. Parallel experiments with Leydig cells in culture gave similar results.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568858 TI - Sialyl SSEA-1 antigen as a carbohydrate marker of human natural killer cells and immature lymphoid cells. AB - The distribution of a carbohydrate antigen, the sialyl SSEA-1 (sialyl Lex-i), in human lymphoid cells was investigated by flow cytometry with a specific monoclonal antibody, MoAb FH-6. We concluded that the lymphocytes positive for the sialyl SSEA-1 antigen present in normal peripheral blood (PB) are natural killer (NK) cells since the positive cells had an NK activity toward K562 cells, and most of the sialyl SSEA-1+ cells were simultaneously positive for Leu-11 (CD 16) and Leu-19. Essentially, no T and B cells, defined by Leu-4 (CD3) and Leu-16 (CD20), were positive for the sialyl SSEA-1 antigen in PB samples taken from healthy donors and patients with disorders unrelated to lymphoid malignancies. Among the malignant lymphoid cells, many sialylated SSEA-1+ cells were observed in large granular lymphocyte (LGL) leukemia cells and some acute lymphoblastic leukemia (ALL) blasts, but not in CLL cells or malignant lymphoma cells. Sialyl SSEA-1 was also positive in some cultured human lymphoid cell lines. We conclude that expression of the sialyl SSEA-1 antigen is strictly limited to a distinct population of NK cells among the mature lymphocytes in normal PB, but the antigen is present in a wide range of immature lymphoblasts of T- and B-cell lineages as well as the NK-cell lineage. The sialyl SSEA-1 antigen disappears from the surface of immature lymphocytes of T- and B-cell lineages during the course of maturation. PMID- 2568859 TI - Helper and suppressor T-cell function in HIV-infected hemophilia patients. AB - T-lymphocyte helper and suppressor functions were assessed in 61 hemophilia patients. Twenty one patients were HIV-negative (Group 1), 27 were HIV-positive without having AIDS-related complex (ARC)/AIDS (Group 2), and 13 had ARC/AIDS (Group 3). T, CD4-positive, or CD8-positive T lymphocytes were cocultured with B lymphocytes and pokeweed mitogen for 6 days and immunoglobulin producing cells were assessed in a reverse hemolytic plaque assay. In HIV-infected patients, T cells as well as the CD4-positive T cell subset exhibited reduced helper (P less than .01, Group 2; P less than .0005, Group 3) and elevated suppressor activity (P less than .02, Group 2; P less than .005, Group 3), whereas no significant difference was found between HIV-negative patients and controls. The number of CD4-positive cells was not correlated with CD4 cell function. CD4-positive cells showed no helper activity (less than 10% of control T cells) in 8/11 (73%), but an excessive suppressor activity (greater than 80% suppression of plaque formation) in 6/11 (55%) Group 3 patients. Our results show that defective helper and elevated suppressor functions of T cells in HIV-infected patients are caused not only by a change in the CD4/CD8 cell counts but also by functional abnormalities of the CD4-positive T-cell subset. These abnormal helper and suppressor functions may play a role in the development of the immunodeficiency state of AIDS patients. PMID- 2568860 TI - Characterization of murine bone marrow and spleen-derived stromal cells: analysis of leukocyte marker and growth factor mRNA transcript levels. AB - Stromal cells are believed to regulate lympho-hematopoiesis through direct cell cell interactions and the release of growth factors. Many questions remain, however, about their lineage derivation and functional heterogeneity. We previously prepared a panel of stromal cell lines from murine spleen and bone marrow and characterized them based on their ability to support lymphocyte growth in long-term cultures. These cells are now compared with respect to their expression of various immunoglobulin superfamily and cytokine genes by Northern blot analysis. These results indicate that although stromal cells appear to be mesodermal in origin, they are not closely related developmentally to the hematopoietic progenitor cells they support. The potential production of at least six cytokines was demonstrated. All clones constitutively expressed mRNA for macrophage colony stimulating factor, interleukin-6, transforming growth factor beta and neuroleukin. The most potent lymphocyte supporting clones also made interleukin 7 constitutively. Previous findings had suggested that these clones responded to exogenous stimuli and this has now been demonstrated in terms of induced expression of IL-6 and G/M-CSF mRNA. Interleukin 6 mRNA levels were markedly upregulated by exposure of cells to LPS, TNF, IL-1, IL-6, IL-7, and EGF. G/M-CSF mRNA levels were "superinduced" by the combination of LPS and cycloheximide, a protein synthesis inhibitor. These responses are similar to ones documented by investigators working with endothelial cells and fibroblasts. Together, these data suggest that stromal cells are a multifunctional component of the lymphopoietic microenvironment and may be active participants in a complex, cytokine-mediated regulatory network. PMID- 2568861 TI - Spectrin Tunis (Sp alpha I/78), an elliptocytogenic variant, is due to the CGG--- TGG codon change (Arg----Trp) at position 35 of the alpha I domain. AB - Spectrin Tunis (Sp alpha I/78) is an alpha l domain variant that causes asymptomatic elliptocytosis in the heterozygote state. It is manifested by a reduction of spectrin dimer self-association and by the development of a major 78 Kd fragment at the expense of the alpha l 80-Kd fragment upon spectrin-limited digestion. Amino acid sequence analysis, following peptide transfer onto Immobilon membranes, showed that the 78-Kd fragment results from a sensitized cleavage after lysyl residue 10. Using a 13.5-kb genomic alpha-spectrin probe and the Xbal, Pvull, and Mspl polymorphic sites detected with this probe, we concluded that spectrin Tunis is associated with the + - + haplotype (in the above order). Twenty mer oligonucleotides, complementary to genomic segments from introns 2 and 3, respectively, were synthesized. We then performed DNA amplification and sequencing. In the two investigated carriers of spectrin Tunis, we found the C----T base substitution of the codon corresponding to position 35 of the alpha l domain (CGG----TGG; Arg----Trp). The mutation lies in the last part of an alpha helix that extends from residues 9 to 44 of partial repeat alpha 1' and is comparable with helix 3 of full repeats 1 to 5. The modified proteolytic site, located 25 amino acid residues upstream, occurs at the beginning of the helix. PMID- 2568862 TI - Sp alpha I/78: a mutation of the alpha I spectrin domain in a white kindred with HE and HPP phenotypes. AB - Limited tryptic digestion of spectrin (Sp) from seven related individuals manifesting hereditary elliptocytosis (HE) or hereditary pyropoikilocytosis (HPP) phenotypes revealed the presence of a novel peptide with a molecular weight of 78 Kd and a concomitant decrease in the alpha I domain (80-Kd peptide), which is the domain involved in the dimer self-association process. Sp from the normal members of this white family exhibited a normal peptide pattern, as compared with controls. The abnormal peptide pattern was associated with a decreased ability of Sp dimer to self-associate. In this kindred in which three generations were available for study, the clinical manifestations were quite variable and ranged from the asymptomatic HE carrier state to hemolytic HE or to severe anemia requiring splenectomy. The severity of the disease appeared to be correlated both with the amount of mutant spectrin (31% to 69%) and with the excess of the Sp dimer found in the membrane (26% to 60%, compared with a normal value of 5.6% +/- 2.2%). Partial amino acid sequencing showed that the alpha I/78-Kd peptide resulted from cleavage at lysine residue 10 of the alpha I/80-Kd domain. Knowledge of the exon/intron structure of cloned genomic DNA encoding the alpha I domain allowed us to amplify in vitro a DNA fragment containing the third exon of the alpha-spectrin gene. The amplified fragment was subcloned and sequenced. A G to T transversion was found in the 39th codon (AGT for AGG), which changed the normal arginine to a serine. Hybridization of amplified DNAs with allele-specific oligonucleotides corresponding to the normal and mutant sequences confirmed the presence of the mutation in three other HE members of the family (the propositus mother, brother, and sister). PMID- 2568863 TI - Improved hematopoiesis in anemic Sl/Sld mice by splenectomy and therapeutic transplantation of a hematopoietic microenvironment. AB - The ability of a clonal hematopoiesis-supportive bone-marrow stromal cell line GBlneor to engraft and alter the microenvironment-induced anemia of Sl/Sld mice was studied. Prior to stromal cell transplantation, Sl/Sld mice received 1 Gy total body irradiation (TBI) and 13 Gy to the right hind limb. Two months after intravenous (IV) injection of 5 x 10(5) GBlneor cells, 54.4% +/- 17.0% donor origin (G418r) colony-forming cells were recovered from the right hind limb of Sl/Sld mice. Long-term bone marrow cultures (LTBMCs) established from GBlneor transplanted mice produced 189.5 CFU-GEMM-forming progenitors/flask over 10 weeks compared with 52.7 +/- 6.2 CFU-GEMM forming progenitors/flask from irradiated nontransplanted Sl/Sld mice. A partial correction of macrocytic anemia was detected 2 months after GBlneor transplantation in splenectomized, irradiated Sl/Sld mice (HgB 7.2 +/- 0.4 g/dL; MCV 68.3 +/- 7.0 fL) compared to splenectomized, irradiated, nontransplanted Sl/Sld mice (HgB 5.5 +/- 1.1 g/dL; MCV 76 +/- 8.5 fL) or control Sl/Sld mice (HgB 5.4 +/- 0.5 g/dL; MCV 82.4 +/- 1.3 fL). Mean RBC volume distribution analysis showed a 2.5-fold increase in percentage of peripheral blood RBCs with MCV less than or equal to 45 fL and confirmed reduction of the MCV in splenectomized-GBlneor-transplanted mice compared to control Sl/Sld mice. A hematopoiesis-suppressive clonal stromal cell line derived from LTBMCs of Sl/Sld mice (Sldneor) engrafted as effectively (43.5% +/- 1.2% G418r CFU-F/limb) as did GBlneor cells (38.3% +/- 0.16% G418r CFU F/limb) to the irradiated right hind limbs of C57Bl/6 mice. LTBMCs established after 2 or 6 months from Sldneor-transplanted mice showed decreased hematopoiesis (182 +/- 12 [2 months] and 3494.3 +/- 408.1 [6 months] CFU-GEMM forming progenitors/flask over 10 weeks) compared to those established from GBlneor transplanted mice (5980 +/- 530 [2 months] and 7728 +/- 607, [6 months] CFU-GEMM progenitors forming/flask). Thus, transplantation of clonal bone-marrow stromal cell lines in vivo can stably transfer their physiologic properties to normal or mutant mice. PMID- 2568864 TI - P-glycoprotein expression in plasma-cell myeloma is associated with resistance to VAD. AB - Tumor cell-associated expression of multidrug resistance (MDR) was quantitated in 22 patients with DNA-aneuploid myeloma using 2-parameter flow cytometry with monoclonal antibody (MoAb) C-219 for the detection of cytoplasmic p-170 and propidium iodide for nuclear DNA content. The proportion of cells expressing p 170 and the intensity of p-170-related fluorescence were determined for each patient. Among the 14 patients treated with vincristine-adriamycin-dexamethasone (VAD), the proportion of p-170-positive cells distinguished sensitive from resistant disease (P less than .01). Among a subgroup of seven patients with MDR analysis available prior to VAD therapy, two subsequent nonresponders had high proportions of C-219-reactive cells. The presence de novo of high proportions of p-170-expressing cells in another still untreated patient and in a further individual with resistance to dexamethasone and interferon (not associated with MDR) warrants systematic analysis of p-170 expression prior to therapy to determine its clinical implications for response to MDR-associated drugs as combined in the VAD regimen. Concurrent MDR expression by aneuploid tumor cells and cells in the diploid subcompartment may represent involvement of diploid cells in the myeloma disease process. PMID- 2568865 TI - Isolation in a single step of a highly enriched murine hematopoietic stem cell population with competitive long-term repopulating ability. AB - A simple procedure is described for the quantitation and enrichment of murine hematopoietic cells with the capacity for long-term repopulation of lymphoid and myeloid tissues in lethally irradiated mice. To ensure detection of the most primitive marrow cells with this potential, we used a competitive assay in which female recipients were injected with male "test" cells and 1 to 2 x 10(5) "compromised" female marrow cells with normal short-term repopulating ability, but whose long-term repopulating ability had been reduced by serial transplantation. Primitive hematopoietic cells were purified by flow cytometry and sorting based on their forward and orthogonal light-scattering properties, and Thy-1 and H-2K antigen expression. Enrichment profiles for normal marrow, and marrow of mice injected with 5-fluorouracil (5-FU) four days previously, were established for each of these parameters using an in vitro assay for high proliferative potential, pluripotent colony-forming cells. When all four parameters were gated simultaneously, these clonogenic cells were enriched 100 fold. Both day 9 and day 12 CFU-S were copurified; however, the purity (23%) and enrichment (75-fold) of day 12 CFU-S in the sorted population was greater with 5 FU-treated cells. Five hundred of the sorted 5-FU marrow cells consistently repopulated recipient lymphoid and myeloid tissues (greater than 50% male, 1 to 3 months post-transplant) when co-injected with 1 to 2 x 10(5) compromised female marrow cells, and approximately 100 were sufficient to achieve the same result in 50% of recipients under the same conditions. This relatively simple purification and assay strategy should facilitate further analysis of the heterogeneity and regulation of stem cells that maintain hematopoiesis in vivo. PMID- 2568866 TI - Biochemical effects of 17 beta-estradiol on UMR106 cells. AB - The effect of 17 beta-estradiol (E) on an osteoblast-like cell line, UMR106, was studied in vitro. The concentrations of transferrin and seven enzymes (gamma glutamyl transferase, alkaline phosphatase, acid phosphatase, lactate dehydrogenase, creatine kinase, alanine aminotransferase and aspartate aminotransferase) were measured in these cells after incubation in culture medium containing either E or the vehicle. E treatment increased five of the seven enzymes and increased the transferrin concentration in the UMR106 cells while simultaneously reducing the proliferation rates. 4-Hydroxytamoxifen, an estrogen antagonist, produced a mild estrogen agonist action on growth rates and enzyme concentrations in the UMR106 cells. When E was present simultaneously, the agonist properties of 4-hydroxytamoxifen were enhanced. These studies show that E enhanced activity of five enzymes and the transferrin content of UMR106 cells after a 2-day incubation. 4-Hydroxytamoxifen enhanced the E effect, illustrating that estrogen antagonists may manifest agonist or antagonist properties depending on the model. These results extend our previous observations showing a direct effect of E in vitro on osteoblast-like cells. PMID- 2568867 TI - Phaeochromocytoma with associated somatostatin production. PMID- 2568868 TI - Soluble interleukin 2 receptor in atopic eczema. AB - OBJECTIVE: To determine whether serum soluble interleukin 2 receptor concentrations are related to disease activity in atopic eczema. DESIGN: Single cohort longitudinal study with controls. SETTING: Outpatient and general medicine departments in secondary referral centre. PATIENTS: Of 15 patients aged 17-57 with severe atopic eczema, all with acute exacerbations of disease, 13 were admitted to hospital and two treated as outpatients until the skin lesions had resolved or greatly improved. Nineteen controls gave single blood samples. INTERVENTIONS: Daily skin dressing with betamethasone valerate (0.025%) and ichthammol paste and tubular dressings. END POINT: Resolution of or considerable improvement in skin lesions. MEASUREMENTS AND MAIN RESULTS: Enzyme linked immunosorbent assays (ELISA) were used to measure serum soluble interleukin 2 receptor concentrations in blood samples taken on admission, at intervals subsequently, and on discharge. Clinical scores of disease activity were also made. Median concentrations on admission were significantly higher (770 U/ml) in the patients than the controls (300 U/ml). Concentrations fell significantly during treatment. In 25 assessments made at different times in 13 patients serum soluble interleukin 2 receptor concentration correlated significantly (R = 0.73) with clinical disease activity. CONCLUSIONS: Cellular immunopathogenic mechanisms contribute to atopic eczema. Immune activation can be measured in atopic eczema by measurements of soluble interleukin 2 receptor, and this should facilitate assessment of response to treatment. PMID- 2568869 TI - L-[35S]cysteic acid selectively detects chloride-dependent L-glutamate transporters in synaptic membrane. AB - Na+-independent L-[35S]cysteic acid (CA) accumulation in rat cortical synaptic membrane was examined. In the absence of Cl-, the accumulation was not observed. Addition of Cl- revealed the accumulation in a dose-dependent manner. Br- and NO3 also did. Ca2+-enhanced the Cl- -dependent accumulation, whereas low concentrations of Na+ reduced it. L-[35S]CA accumulation was inhibited by quisqualate, L-glutamate (L-Glu), L-cysteine sulfinate, D,L-homocysteic acid and D,L-2-amino-4-phosphonobutyrate (D,L-APB) potently. L-CA inhibited L-[3H]Glu accumulation in synaptic membrane in the presence of Cl-. The maximal inhibition of L-CA was equal to that of D,L-APB, but L-CA did not inhibit L-[3H]Glu accumulation in the absence of Cl-. These results show that L-[35S]CA selectively detects the Cl- -dependent L-Glu transporters in synaptic membrane. PMID- 2568870 TI - Responses of isolated white perch horizontal cells to changes in the concentration of photoreceptor transmitter agonists. AB - Current and voltage responses elicited by increasing or decreasing the concentration of L-glutamate or its analog kainate around isolated cone horizontal cells were measured with patch pipettes using the whole cell recording configuration. Application of these photoreceptor transmitter agonists induced inward currents in voltage-clamp experiments (for negative holding potentials) and depolarizing responses in current-clamp experiments. Continuous exposure to either drug produced inward currents which were maintained for as long as superfusion with the drugs continued. Reducing the concentration of the agonists by pressure ejection of pulses of drug-free Ringer's solution onto the cells completely turned off the drug-induced currents. Under current-clamp conditions, pulses of control Ringer's elicited hyperpolarizing responses of large amplitude (40-80 mV). The data demonstrate the ability to simulate in vitro the horizontal cell's photoresponses and thus support the use of cultured cells as a model system for studying horizontal cell physiology and pharmacology. PMID- 2568871 TI - Somatostatin presynaptically inhibits transmitter release in feline parasympathetic ganglia. AB - Intracellular recordings were made from neurons in cat parasympathetic ciliary ganglia in vitro. Somatostatin (30 nM-3 microM) reduced the amplitude of excitatory postsynaptic potentials (EPSPs), whereas the peptide did not affect acetylcholine (ACh)-induced depolarizations. Thus somatostatin depressed the EPSPs without changing the postsynaptic sensitivity to ACh. The inhibitory action of somatostatin on the EPSPs was passed off even in the presence of the peptide at concentrations higher than 100 nM. When paired stimuli at an interval of 50 ms were applied to preganglionic nerves, the second EPSP was facilitated, being larger in amplitude than the first one; this facilitation was reversibly inhibited in the presence of the peptide. Somatostatin reversibly reduced the frequency of spontaneous EPSPs without appreciably changing their mean amplitude. All of these results indicate that somatostatin may presynaptically reduce the amount of ACh released. The mechanism underlying this action was discussed. PMID- 2568872 TI - Synaptic potentials in the rat neostriatum in dissociated embryonic cell culture. AB - Neostriatal cells of embryonic days 19-21 were grown in dissociated cell culture. To test whether the cultures contained predominantly neostriatal cells, a glyoxylic acid staining procedure was used which, after dopamine loading, stained neostriatal cells but not neurons of embryonic neocortical tissue. Whole cell current clamp recording was performed in the neurons after 1-2 weeks in cell culture. Although cells could be driven to discharge by direct depolarization, spontaneous activity was low. All cells responded to gamma-aminobutyric acid (GABA) (0.1-0.5 mM), and the majority of them responded to glutamate (Glu) (0.1 mM). Only about 50% were depolarized by acetylcholine (ACh) (0.1-0.5 mM). Atropine (1-10 microM) did not block this depolarization. Barrages of postsynaptic potentials (PSPs) were induced by applications of Glu or ACh, even if the neuron under observation was not depolarized. All PSPs were depressed by bicuculline (50 microM), indicating their mediation by GABAergic receptors. Exclusively GABAergic PSPs were also observed in cultures raised in the presence of nerve growth factor. The study indicates that neostriatal cells form GABAergic, but not excitatory cholinergic synapses when cultured at this embryonic age under our conditions, resembling the pattern of development observed in slices obtained from neonatal rats. PMID- 2568873 TI - Effects of the noradrenergic neurotoxin DSP-4 on luteinizing hormone levels, catecholamine concentrations, alpha 2-adrenergic receptor binding, and aromatase activity in the brain of the Japanese quail. AB - Previous investigations have established that DSP-4 reliably enhances the activating effects of testosterone on copulatory behavior in adult male quail. In the present study, we wanted to clarify the neurochemical changes that parallel these behavioral effects and to determine whether DSP-4 also affects non behavioral steroid-dependent sexually dimorphic reproductive processes. We first showed using the Palkovits microdissection technique combined with assay by high performance liquid chromatography (HPLC) that DSP-4 specifically depletes norepinephrine in several nuclei of the brain such as the medial preoptic nucleus, the ventromedial nucleus of the hypothalamus or the intercollicular nucleus but leaves intact the noradrenergic innervation in other areas such as the infundibulum or nucleus accumbens. Other amines such as dopamine and serotonin were not affected by the drug. Surprisingly DSP-4 did not decrease the binding of tritiated p-aminoclonidine in any of the brain areas which were studied by quantitative autoradiography. This suggests that most of the alpha 2 adrenergic receptors are located at the postsynaptic level but alternative interpretations are discussed. Testosterone treatment of castrated birds specifically reduced the density of alpha 2-adrenergic receptors in the dorsal infundibulum and in the medial mammillary nucleus. The possible relations of this receptor change to the control of luteinizing hormone (LH) secretion are discussed. Finally it was shown that DSP-4 treatment decreases plasma LH levels (which reveals the stimulatory effect of norepinephrine on LH secretion) but increases the testosterone-induced aromatase activity in the preoptic area. This latter effect could be one of the mechanisms by which DSP-4 potentiates copulatory behavior in testosterone-treated quail. PMID- 2568874 TI - Characterization of striatal neurons expressing high levels of glutamic acid decarboxylase messenger RNA. AB - Two types of labelled cells are detected in sections of rat and mouse striata processed for in situ hybridization histochemistry with 35S-radiolabelled RNA probes complementary to the messenger RNA (mRNA) encoding glutamic acid decarboxylase (GAD), the synthesis enzyme for gamma-aminobutyric acid (GABA): numerous lightly, and fewer very densely labelled neurons. In order to determine whether the densely labelled cells correspond to the striatal somatostatinergic neurons with which they share morphological characteristics, the presence of GAD mRNA was examined in brain sections processed successively for dihydronicotinamide adenine dinucleotide phosphate (NADPH) diaphorase histochemistry, a marker of striatal somatostatinergic neurons, and in situ hybridization histochemistry. In addition, the distribution of GABAergic interneurons was analyzed with regard to striatal compartments (striosomes) indicated by patches of dense opiate binding sites. The results show that NADPH diaphorase activity and GAD mRNA do not co-exist in striatal neurons. Furthermore, in contrast to the somatostatinergic neurons which are almost exclusively located in the extrastriosomal matrix, densely labelled GAD cells were present both in the striosomes and the matrix, further suggesting that GABAergic and somatostatinergic neurons form two distinct interneuronal systems in the striatum of rats and mice. PMID- 2568875 TI - Collateral sprouting of somatostatin-immunoreactive axons after partial deafferentation of the central nucleus of the rat amygdala. AB - These experiments utilize a paradigm developed to study plastic responses of peptidergic neurons in a discrete brain area following deafferentation. The central nucleus of the amygdala (CNA) is richly innervated by somatostatin immunoreactive (SS-I) terminal axons. In the course of preliminary light microscopic (LM) investigations by this laboratory, changes were observed in the density of presumed SS-I terminals in the rat CNA after lesioning the medial input. The LM finding of increased density of presumed SS-I terminals in the CNA at the 10-day post-lesion stage underscored the need for a quantitative electron microscopic (EM) study of the SS-I components, including an evaluation of synaptic events at different survival periods. At the 3-day post-lesion stage, EM examination showed degenerating axons in the lesioned CNA, many already engulfed by astrocytes. None of the degenerating profiles were SS-I, supporting the view that the lesion did not interrupt, to any significant extent, SS-I axons entering the nucleus. EM surveys of the 10-day post-lesion material demonstrated that degenerated profiles had almost completely disappeared. Numbers of SS-I axon terminals, particularly of smaller-sized profiles, were increased by 22% over control value. Synaptic frequency was decreased by 16% below control value. Numbers of SS-I terminals making synapses were increased 3.4% above control value. At the 30-day post-lesion stage, the total number of SS-I terminal axons had increased 86% over controls, whereas the synaptic frequency had decreased by about a third below controls. The absolute number of SS-I terminals engaging in synapses had increased by 24% over controls. The 90-day post-lesion CNA showed a further increase in the number of SS-I axon profiles: 136% over control value. The synapse-to-axon ratio (synaptic frequency) of 27% was similar to that observed for the CNA from the unlesioned side or from unoperated animals. At this stage the number of SS-I synapses had increased by 135% over controls. This model presents many possibilities for studying neuroplasticity, particularly involving peptidergic neurons of the central autonomic nervous system. PMID- 2568877 TI - Selective effects of serotonin upon excitatory amino acid-induced depolarizations of Purkinje cells in cerebellar slices from young rats. AB - The effects of serotonin on responses induced in Purkinje cells (PCs) by microiontophoretic administration of excitatory amino acids (EAAs) in their dendritic fields were tested in vitro by extracellular recording and by single electrode voltage clamp methods in cerebellar slices from rats aged 16-22 days. Serotonin diminished excitations produced by glutamate (Glu) and quisqualate (Quis) selectively, those caused by N-methyl-D-aspartate (NMDA) being affected much less. These suppressions of Glu- and Quis-induced responses generally occurred without there being any effect on intrinsic membrane properties of PCs, although on occasion serotonin increased membrane conductance slightly and/or induced an outward current in the recorded cells. All these effects of serotonin were maintained in the presence of tetrodotoxin and reversed upon removal of the amine. On the few occasions when serotonin enhanced Quis-induced responses, the effect was mimicked by ejection from a control solution of saline, made up at the same pH as the drug solution of serotonin. PMID- 2568876 TI - Thalamocortical synapses with identified neurons in monkey primary auditory cortex: a combined Golgi/EM and GABA/peptide immunocytochemistry study. AB - The objective of this study was to identify neurons in layer IV of the monkey primary auditory cortex (area KA) that are postsynaptic to thalamocortical axon terminals. Thalamocortical axon terminals were labeled by lesion-induced degeneration; neurons postsynaptic to these afferents were labeled by the Golgi/EM method followed by postembedding immunocytochemistry. Five of the six non-pyramidal neurons examined received synapses from thalamocortical axon terminals. All of these cells were immunoreactive for gamma-aminobutyric acid (GABA). One of the cells stained also with an antiserum to somatostatin, and another for cholecystokinin. None of the cells examined were immunoreactive to substance P, and in no instance were two different peptides co-localized within the same GABA-positive neuron. PMID- 2568878 TI - Effects of xylazine on cerebrospinal fluid catecholamines in the rhesus monkey. AB - The i.v. administration of xylazine, a potent, selective alpha 2-adrenergic receptor agonist, resulted in a 76% decrease in cerebrospinal fluid (CSF) norepinephrine in chair-adapted rhesus monkeys. A significant decrease was observed within 1.5 h of administration and continued through the 3 h course of sampling. Dopamine was maximally decreased by 24% at 1.5 h. Epinephrine was not significantly decreased following xylazine administration. These data suggest that norepinephrine release into monkey CSF, as an index of central or peripheral norepinephrine turnover, is more sensitive to alpha 2-adrenergic agonists than in CSF dopamine or epinephrine. PMID- 2568879 TI - L-beta-methylaminoalanine inhibits [3H]glutamate binding in the presence of bicarbonate ions. AB - We examined the ability of the neurotoxin, L-beta-methylaminoalanine (L-BMAA), to inhibit [3H]glutamate binding to rat brain synaptic junctions. In a tris(hydroxymethyl)aminomethane acetate buffer, L-BMAA did not affect [3H]glutamate binding (IC50 greater than 10 mM). However, in the presence of ammonium bicarbonate (20 mM) L-BMAA blocked [3H]glutamate binding with an IC50 of 1 mM. This inhibition was not caused by ammonium ion since other ammonium salts were inactive. Furthermore, identical inhibition was obtained in the presence of potassium bicarbonate. Bicarbonate ion did not alter the ability of N-methyl-D aspartic acid to block glutamate binding. These results indicate that bicarbonate ion is required for the interaction of L-BMAA with the glutamate receptor and may account for the observation that beta-methylaminoalanine is neurotoxic in vitro only in the presence of bicarbonate. PMID- 2568880 TI - Somatostatin inhibition of adenylate cyclase activity in different brain areas. AB - Somatostatin receptors have been identified in different brain areas but the characterization of their postreceptor effect is still lacking. In this study we analyze the somatostatin effect on adenylate cyclase activity in different brain regions, namely frontal cortex, striatum, hypothalamus and hippocampus. Somatostatin inhibited basal adenylate cyclase activity in all brain areas, being maximally effective in the frontal cortex (-42%). Moreover, somatostatin inhibited both dopamine- and norepinephrine-stimulated adenylate cyclase activity in the examined cerebral regions showing a higher effectiveness than in basal conditions. VIP stimulation of adenylate cyclase was also reduced by somatostatin. The peptide inhibited by 50% forskolin-stimulated (10 nM to 10 microM) enzyme activity in frontal cortex and hypothalamus (in hippocampus the inhibition reached only -25%) showing a non-competitive pattern of inhibition. Finally, pertussis toxin pretreatment abolished the somatostatin inhibition of forskolin-stimulated frontal cortex adenylate cyclase activity. These results show that brain somatostatin receptors are coupled in an inhibitory way with adenylate cyclase enzyme that may represent one of the postreceptor mechanisms mediating the somatostatin modulation of brain functions. PMID- 2568881 TI - The effect of an increase in inhaled allergen dose after terfenadine on the occurrence and magnitude of the late asthmatic response. AB - We have attempted to use a potent and selective histamine H1-receptor antagonist terfenadine to allow a larger dose of allergen to be administered to previous single early responders to investigate if an increased dose of allergen could induce a late asthmatic response. Pre-treatment with 180 mg of terfenadine enabled a geometric mean increase in allergen dose of 4.12-fold to be inhaled by eight atopic subjects with mild asthma, who initially were classified as single early responders, with maximal fall in FEV1 3-8 hr after allergen challenge (Lmax) of less than 15% from baseline value. The magnitude of early asthmatic response was similar to that obtained on the control day when allergen challenge was performed in the absence of terfenadine. Two subjects were converted to dual responders with Lmax of 23.1 and 24.3%, which occurred with a 32- and 65-fold increase in allergen dose respectively, and a 6- and 4.9-fold decrease in non specific airways responsiveness measured as the cumulative provocative concentration of methacholine that caused a 20% fall in FEV1 from baseline. The remaining six subjects failed to achieve an Lmax of greater than 10% even with a 1.29-2.66-fold increase in allergen dose. For the group as a whole an increase in allergen dose was associated with an increase in overall bronchoconstrictor response 3-8 hr after challenge. These results indicate that it is possible to induce a late asthmatic response in a subject who previously demonstrated only an early response by increasing the dose of allergen inhaled. PMID- 2568882 TI - The epidemiology of foot and ankle injuries in sports. AB - A 6 1/2-year review of 16,754 injuries seen in a multispecialty sports medicine clinic found that 25 per cent of the 12,681 injuries in the top 19 sports occurred at the ankle and foot. The percentages of foot and ankle injuries varied substantially from sport to sport, as did the proportion of sprains versus overuse injuries at each location. An appreciation of the patterns and numbers of injuries presenting can be helpful in patient management and can aid the planning of both clinical investigations and educational programs. PMID- 2568883 TI - The rehabilitation of overuse foot injuries in athletes and dancers. AB - Overuse injuries in athletes and dancers provide a challenge to the diagnostic acumen and rehabilitative and preventive skills of the medical community. The challenge is intensified when the primary lesion is located in the foot, whose levers, arches, and pulleys are responsible for absorbing shock, as well as converting a mobile adapter into a rigid lever for propulsion. A thorough knowledge of these active and passive mechanisms is essential for the appropriate and expeditious management of overuse syndromes. Sports and dance medicine health practitioners must also explore and comprehend the kinesiology of the sports and/or dance activity prevalent among their clientele. Furthermore, in order to determine the etiology and make judicious recommendations, clinicians must familiarize themselves with the density and inclines of popular running and dancing surfaces within their referral area. Rehabilitation of the foot requires a multifaceted approach. Physical therapists dealing with these problems need to develop expertise in manual techniques, along with clinical competency in laser, electrical, and thermal modalities. Reduction of predisposing factors is of utmost importance in the successful management of overuse injuries, and often requires greater skill and attention than does the treatment of the primary lesion. PMID- 2568885 TI - Elevated levels of prostaglandin E2 in the liver of rats fed a choline deficient diet: possible involvement in liver tumor promotion. AB - The effect of feeding a choline deficient (CD) diet, an efficient liver tumor promoting regimen, on the prostaglandin metabolism in the liver of male Sprague Dawley rats was investigated. The possible biological significance of the alteration was examined using hypolipidemic peroxisome proliferators and modifiers of prostaglandin metabolism such as indomethacin and menhaden oil in the short term assay of the induction of enzyme altered foci in the liver. A CD diet, when fed for 10-30 days, induced 2-2.5 times increases in the levels of prostaglandin E2 (PGE2) in the liver, while the hypolipidemic peroxisome proliferators, 4-chloro-6(2,3-xylidino) pyrimidinylthio(N-hydroxyethyl)-acetamide (BR931) and di(2-ethylhexyl)-phthalate (DEHP), markedly reduced the levels of this metabolite. The addition of BR931 or indomethacin to a CD diet suppressed the diet-induced elevations of PGE2 and a substitution of fats in a CD diet with menhaden oil had the same effect. Furthermore, both indomethacin and menhaden oil added to a CD diet suppressed the induction of gamma-glutamyltranspeptidase positive hepatocyte foci in the liver of rats initiated with a single dose of diethylnitrosamine after 8 weeks of the dietary promotion. The results suggest that altered prostaglandin metabolism may be involved in the liver tumor promoting effect of a CD diet. PMID- 2568884 TI - Loss of heterozygosity on 3p in a renal cell carcinoma in von Hippel-Lindau syndrome. AB - A renal cell carcinoma with an unbalanced t(X;3) in a patient with von Hippel Lindau (VHL) syndrome has previously been reported. This rearrangement suggested loss of genetic material from the short arm of chromosome 3, which we are now able to confirm by restriction fragment length polymorphism analysis of tumor DNA using polymorphic probes derived from 3p. The VHL gene has recently been mapped to 3p, therefore loss of this region in this VHL-related renal cell carcinoma may have cogent significance for tumor development in this interesting cancer predisposing syndrome. PMID- 2568886 TI - Augmentation of the generation of lymphokine-activated killer cells after a single dose of mitomycin C in cancer patients. AB - The effect of mitomycin C administration on the generation of cytotoxic cells, induced by in vitro activation of peripheral blood mononuclear cells (PBM) with interleukin-2, was studied in patients with various carcinomas. The ability of PBM to generate lymphokine-activated killer (LAK) cell activity against Raji cell targets was significantly augmented 5 and 7 days after a single intravenous dose of 12 mg/m2 mitomycin C, when compared to that of PBM obtained before mitomycin C injection. Further, LAK cell activity against autologous tumor cells was also significantly increased after the drug administration. The distribution of lymphocyte subsets exhibited a significant increase in the percentage of CD3+ cells after injection, with the elevation of the CD4/CD8 ratio. Furthermore, the proportion of the CD4+ Leu8+ subpopulation, which identifies inducers of suppression, was significantly reduced. Thus, the decrease in the proportion of suppressor-inducer subsets of PBM might be at least partially, responsible for the augmented generation of LAK cells after mitomycin C administration. PMID- 2568887 TI - Expression of the v-erbA product, an altered nuclear hormone receptor, is sufficient to transform erythrocytic cells in vitro. AB - We investigated the effect of the v-erbA oncogene product, an altered thyroid hormone receptor, in chicken erythrocyte progenitor cells. Bone marrow cells were infected with a retrovirus vector (XJ12) carrying the v-erbA gene in association with the neoR gene. XJ12-infected erythrocyte progenitor cells gave rise to G418 resistant clones. Some were composed of blast cells identified as transformed CFU Es blocked in their differentiation. These cells could be grown in culture for at least 25 generations and required anemic chicken serum as a source of erythropoietic growth factors. XJ12 can infect erythrocyte progenitor cells in vivo but is not sufficient to induce erythroleukemia. These data suggest that the activation of a nuclear hormone receptor might represent one step toward the development of neoplasms. PMID- 2568888 TI - Synergistic interaction of p185c-neu and the EGF receptor leads to transformation of rodent fibroblasts. AB - The protein product of the rodent neu oncogene, p185neu, is a tyrosine kinase with structural similarity to the epidermal growth factor receptor (EGFR). Transfection and subsequent overexpression of the human p185c-erbB-2 protein transforms NIH 3T3 cells in vitro. However, NIH 3T3 cells are not transformed by overexpressed rodent p185c-neu. NIH 3T3 transfectants overexpressing EGF receptors are not transformed unless incompletely transformed. Several groups have recently demonstrated EGF-induced, EGFR-mediated phosphorylation of p185c neu. During efforts to characterize the interaction of p185c-neu with EGFR further, we created cell lines that simultaneously overexpress both p185c-neu and EGFR and observed that these cells become transformed. These observations demonstrate that two distinct, overexpressed tyrosine kinases can act synergistically to transform NIH 3T3 cells, thus identifying a novel mechanism that can lead to transformation. PMID- 2568890 TI - Fc receptor isoforms exhibit distinct abilities for coated pit localization as a result of cytoplasmic domain heterogeneity. AB - Mouse macrophages and lymphocytes express two distinct isoforms of a single class of Fc receptor for IgG. The macrophage isoform (FcRII-B2) is identical to the lymphocyte isoform (FcRII-B1) except for an inframe insertion in the cytoplasmic tail of FcRII-B1 that increases its length from 47 to 94 amino acids. To determine the functional significance of this cytoplasmic domain variation, presumably the result of alternative mRNA splicing, we expressed both isoforms in receptor-negative fibroblasts. While FcRII-B2 mediated the efficient ligand internalization and delivery to lysosomes, endocytosis via FcRII-B1--and via a tailminus mutant--was relatively inefficient. This difference reflected the inability of FcRII-B1 (and the tailminus mutant) to accumulate in clathrin-coated pits. Thus, the FcRII-B2 cytoplasmic tail contains a domain needed for accumulation in coated pits, and this domain is disrupted by the 47 amino acid insertion in FcRII-B1. PMID- 2568889 TI - Structure of the gene of tum- transplantation antigen P91A: the mutated exon encodes a peptide recognized with Ld by cytolytic T cells. AB - Mutagen treatment of mouse P815 tumor cells produces immunogenic mutants that express new transplantation antigens (tum- antigens) recognized by cytolytic T cells. We found that the gene conferring expression of tum- antigen P91A contains 12 exons, encoding a 60 kd protein lacking a typical N-terminal signal sequence. The sequence shows no significant similarity with sequences in current data bases. A mutation that causes expression of the antigen is located in exon 4; it is the only apparent difference between the normal and the antigenic alleles. A short synthetic peptide corresponding to a region of exon 4 located around this mutation makes P815 cells sensitive to lysis by anti-P91A cytolytic T cells. The mutation creates a strong aggretope enabling the peptide to bind the H-2 Ld molecule. Several secondary tumor cell variants that no longer express tum- antigen P91A were found to carry deletions in the gene. PMID- 2568891 TI - Craniofacial abnormalities induced by ectopic expression of the homeobox gene Hox 1.1 in transgenic mice. AB - Hox-1.1 is a murine homeobox-containing gene expressed in a time- and cell specific manner during embryogenesis. We have generated transgenic mice that ectopically express Hox-1.1 from the chicken beta-actin promoter. In these mice Hox-1.1 expression was changed to an almost ubiquitous pattern. Ectopic expression of Hox-1.1 leads to death of the transgenic animals shortly after birth and is associated with multiple craniofacial anomalies, such as cleft palate, open eyes at birth, and nonfused pinnae. This phenotype is similar to the effects seen after systemic administration of retinoic acid during gestation. This suggests that retinoic acid embryopathy and the specific developmental defects caused by ectopic expression of a potential developmental control gene share a common pathogenic mechanism. PMID- 2568892 TI - Interspecies differences in pharmacokinetics of an antiallergic agent, 1-(2 ethoxyethyl)-2-(hexahydro-4-methyl-1,4-diazepin-1-yl) benzimidazole difumarate (KG-2413) after intravenous administration to rats, guinea pigs and dogs. AB - Interspecies differences in the pharmacokinetics of an antiallergic agent, 1-(2 ethoxyethyl)-2-(hexahydro-4-methyl-1,4-diazepin-1-yl)benzimid azole difumarate (KG-2413) after intravenous administration were investigated in rats, guinea pigs and dogs. The disappearance of unchanged KG-2413 base was described by biexponential curves in all three animal species. The areas under the plasma concentration-time curves (AUC) in rats, guinea pigs and dogs were 218, 421 and 369 ng.h/ml, respectively, at a dose of 2 mg/kg. The volume of distribution in rats was comparable to that in dogs, and was about three times greater than that in guinea pigs. This might be due to the difference in the unbound fractions of KG-2413 base in plasma (fu), that is, the values of fu in rats, dogs and guinea pigs were 0.607, 0.603 and 0.189, respectively. The first-order elimination rate constant from the central compartment (kcl) in dogs were smaller than those in rats and guinea pigs. Total body clearances (CLtot) of KG-2413 base in rats and guinea pigs were comparable to the hepatic blood flow rate (Q) of each animal. Assuming that KG-2413 base is only eliminated in the liver, relatively rapid metabolism of KG-2413 base occurred in the liver of rats and guinea pigs. In dogs, extrahepatic elimination was suggested because the value of CLtot seemed to be greater than that of Q. PMID- 2568893 TI - Differing circadian patterns of symptom onset in subgroups of patients with acute myocardial infarction. AB - Circadian variation of the onset of acute myocardial infarction has been noted in many studies and may carry important pathophysiologic implications. However, only a few previous studies have attempted subgroup analyses. In 4,796 patients with documented acute myocardial infarction, the time of symptom onset was recorded. As in other studies, the peak of onset occurred in the morning from 6:01 AM to 12:00 noon, and 28% of the population (1.16 times the average percentage for the other time periods) experienced symptom onset in that period (p less than 0.001). There was a second, lower peak (25%) in the evening between 6:01 PM and 12:00 midnight, which was also observed in some previous studies. We sought to determine whether or not the presence of subgroups with specific clinical characteristics would exhibit different patterns and thereby contribute to these peaks in the overall population. In patients with a history of congestive heart failure (n = 606) or with non-Q wave infarction (n = 832), a pronounced peak (29%) occurred only in the evening. Two nearly equal peaks were observed in patients older than 70 years of age (n = 1,422), smokers (n = 2,057), diabetics (n = 767), women (n = 1,213), and patients taking beta-blocking drugs (n = 847). Finally, in patients with a previous myocardial infarction (n = 1,104), no peaks were observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568894 TI - Distribution and morphology of tyrosine hydroxylase-immunoreactive neurons in the developing mouse retina. AB - An antibody to tyrosine hydroxylase (TH), the rate-limiting enzyme in the production of catecholamines, was used to examine the morphology and distribution of catecholaminergic neurons in whole-mounted retinae of the developing and adult mouse. At adulthood TH-labeled cell bodies were located in the inner nuclear layer, stratifying mainly at the border to the inner plexiform layer (IPL). Few processes were found in the middle of the IPL. The majority of all TH-labeled cells also extended processes towards the outer plexiform layer and thus are interplexiform cells; the rest were considered to be amacrine cells. The TH positive neurons were regularly distributed throughout the adult mouse retina. During development, the first TH-immunoreactive cells were observed by postnatal day 6 (P6) and most of them were present after the third postnatal week. The dendrites in the IPL only acquired varicosities after the eyes opened at P15. Biochemical measurements of the endogenous catecholamine content showed that at all developmental stages only dopamine was detectable, suggesting that the TH labeled cells represent dopaminergic neurons. The content of dopamine was low before P6 and continuously increased during the following days. A strong increase in dopamine was observed during the time when varicosities formed. PMID- 2568896 TI - [Catecholaminergic systems in the amygdaloid complex of SDAT and aged controls: tyrosine hydroxylase immunohistochemistry]. AB - Catecholaminergic systems in the amygdaloid complex in patients with senile dementia of Alzheimer type (SDAT) and aged controls have been studied with tyrosine hydroxylase (TH) immunohistochemistry. TH-immunoreactivity was found in fibers in all subnuclei of human amygdala, although varying in density in the subnuclei. The most dense catecholaminergic innervation was observed in the central, basal, and cortical nuclei of amygdala. The prominent finding in the amygdaloid complex of SDAT was that swollen and bulbous TH-immunoreactive neurites were found in association with neuritic plaques, which have not, rarely if any, been found in controls. The numerical density of neurites was mostly in parallel with that of TH-immunoreactive fibers except for the central nucleus. Thus, it is suggested that a class of populations of TH-immunoreactive neurons are selectively affected in the amygdaloid complex in SDAT. PMID- 2568895 TI - [Pathology of the peripheral nervous system in polyarteritis nodosa: a clinico pathological study of two autopsy cases]. AB - Peripheral nerves from the two autopsied cases with polyarteritis nodosa were pathologically examined. Case 1 was a seventy-year-old female, complaining of numbness and weakness of the extremities. Sensory deficits in all modality with induced dysesthesia were distributed as a mode of overlapping mononeuritis multiplex in the distal portion of extremities. Motor involvement was also noted in extremities in disarray-accentuated pattern. Extensive steroid hormone therapy was performed and remarkable improvement in clinical sign was temporarily obtained. Relapse with bowel involvement was a cause of the death. In the post mortem examination, central fascicular degeneration with the loss of large myelinated fibers were seen at the middle portion of upper limbs in median nerve and at the lower mid-thigh in sciatic nerve. In the distal portion of those nerves, diffuse extensive loss of large myelinated fibers in the fasciculus were observed. Some of small thin myelinated fibers in the central fasciculus or distal portion of nerves were thought to be regenerated in nature. Accumulation of cell organelles in the axonal swellings were frequently occurred in the proximal to the ischemic site. Case 2 was a seventy five-year-old female with a both motor and sensory involvement in the distal extremities. Right pulmonary effusion and extensive subcutaneous hemorrhage and necrotic gangrene in the leg was also noted. Post-mortem examination of peripheral nerves revealed a presence of central fascicular degeneration in the distal sciatic nerves and nearly complete loss of myelinated fibers in the distal nerves. Ventral and dorsal roots and dorsal root ganglia were well preserved.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568897 TI - Serotonin syndrome versus neuroleptic malignant syndrome as a cause of death. PMID- 2568898 TI - A critical review of continuous infusion H2 receptor therapy. AB - Recent studies suggest that continuous infusions of H2-receptor antagonists may be more effective than intermittent iv therapy to control gastric pH for the prevention of stress ulcers. Infusions of cimetidine, ranitidine, and famotidine have been shown either to have a beneficial clinical effect, to control gastric acid secretion, to maintain gastric pH greater than 4, or to have pharmacokinetic properties similar to intermittent doses. In addition, some evidence suggests that continuous infusions may permit the use of lower doses, leading to cost savings. However, as yet there have been no trials that directly compare continuous and intermittent regimens and show improved outcomes with a continuous regimen. Since candidates for parenteral administration of H2-receptor antagonists are most common in the ICU, several practical issues must be addressed. Patients who receive total parenteral nutrition (TPN) may be suitable candidates for continuous infusions of H2-blockers since these agents can be given in the same container as TPN solutions. These patients also usually have a parenteral access site and infusion pump dedicated to TPN administration. In other patients, drug incompatibilities, limited iv access lines, or a lack of infusion pumps may require frequent interruptions of the infusion in order to administer additional medications, which may lead to a loss of gastric pH control. In most patients, administration of an H2-blocker regimen which maintains consistent pH control after intermittent administration may be the most practical method by which to administer these agents. PMID- 2568899 TI - Calcium channel blockers as inhibitors of drug metabolism. PMID- 2568901 TI - [Polymorphic changes in cellular phenotype during hyperexpression of the NEU proto-oncogene]. PMID- 2568902 TI - Identification and kinetic characterization of acetylator genotype-dependent and independent arylamine carcinogen N-acetyltransferases in hamster bladder cytosol. AB - Recent studies from our laboratory have shown relatively high levels of polymorphic N-acetyltransferase (NAT)(EC 2.3.1.5) activity toward carcinogenic arylamines in urinary bladder cytosol of humans and in the inbred hamster model of the N-acetylation polymorphism. The expression of this polymorphism is of interest because of the higher incidence of bladder cancer among human slow acetylators with documented exposures to arylamine bladder carcinogens. In this study, arylamine NAT activity was partially purified and characterized in inbred hamster urinary bladder cytosols of defined acetylator genotype. Acetylator gene dose response relationships were observed for the N-acetylation of p-aminobenzoic acid, p-aminosalicyclic acid, and the arylamine carcinogens 2-aminofluorene, 4 aminobiphenyl, and beta-naphthylamine in hamster bladder cytosol. Partial purification of hamster bladder cytosol by anion-exchange fast protein liquid chromatography yielded two NAT isozymes that catalyzed the N-acetylation of each of the arylamine substrates. The catalytic activity of the first isozyme was acetylator genotype-dependent (polymorphic), whereas the second isozyme appeared to be acetylator genotype-independent (monomorphic). Catalytic activities between homozygous rapid, heterozygous, and homozygous slow acetylator genotypes were compared with respect to both initial rates and apparent maximum velocities. Comparison of homozygous rapid and slow acetylator bladder cytosol showed that the apparent Vmax for 2-aminofluorene NAT activity was significantly higher in rapid than slow acetylators (6-fold in cytosol, 50-fold in the polymorphic NAT isozyme). These results suggest a key role for a polymorphic NAT isozyme, regulated by the acetylator genotype and expressed in urinary bladder cytosol, in the initiation of bladder cancer via arylamine carcinogens. PMID- 2568900 TI - Regional differences in concentrations of regulatory peptides in human colon mucosal biopsy. AB - The study was undertaken to examine regional differences in the concentrations of five regulatory peptides in the human colonic mucosa. Biopsies were obtained during routine colonoscopy from 33 patients whose colonic mucosa was macroscopically and histologically normal. Regulatory peptides were extracted, and measured by specific radioimmunoassays. Concentrations of three peptides that are present predominantly in endocrine cells within colonic mucosa increased significantly towards the rectum: Mean concentrations of peptide YY, enteroglucagon, and somatostatin were about three times greater in the rectum than in the cecum. However, concentrations of two peptides that are present in mucosal nerve fibers diminished significantly towards the rectum: Mean rectal concentrations of vasoactive intestinal peptide and peptide histidine methionine were both about 0.6 of mean cecal concentrations. Concentrations of all five peptides were lower in biopsies taken from colonic polyps than in normal colonic mucosa. Regional differences in colonic mucosal concentrations of regulatory peptides probably reflect differences in the physiological functions of different parts of the colon. PMID- 2568903 TI - Kinetic characterization of acetylator genotype-dependent and -independent N acetyltransferase isozymes in homozygous rapid and slow acetylator inbred hamster liver cytosol. AB - Acetyl-coenzyme A (AcCoA)-dependent arylamine N-acetyltransferase (NAT) activity (EC 2.3.1.5) was examined in liver cytosol derived from homozygous rapid acetylator (Bio. 87.20) and homozygous slow acetylator (Bio. 82.73/H) Syrian inbred hamsters. Expression of NAT activity toward p-aminobenzoic acid (PABA), 2 aminofluorene (AF), and 4-aminobiphenyl (ABP) was acetylator genotype-dependent, whereas N-acetyltransferase activity toward isoniazid was acetylator genotype independent. Two isozymes of NAT activity were partially purified by anion exchange fast protein liquid chromatography from the hepatic cytosol of both homozygous rapid and homozygous slow acetylator hamsters. The first eluting NAT isozyme exhibited a polymorphic expression toward AF, ABP, and PABA although the second eluting NAT isozyme exhibited a monomorphic expression across acetylator genotypes toward the same substrates. Determination of Michaelis-Menten kinetic constants in hepatic cytosol of homozygous rapid and slow acetylator hamsters suggests that PABA, AF, and ABP NAT activities were acetylator genotype-dependent because of catalysis by polymorphic NAT isozyme that is both an apparent Km and Vmax variant, whereas, the acetylator genotype-independent expression of isoniazid NAT activity appeared to result from catalysis via a common monomorphic NAT isozyme in both acetylator genotypes. Additional kinetic studies on the partially purified NAT isozymes of homozygous rapid and slow acetylator hamster liver confirmed that the polymorphic NAT isozyme exhibited a substantially higher apparent maximum velocity in homozygous rapid acetylators than slow acetylators toward PABA, AF, and ABP as well as acetylator genotype-related differences in the apparent Km toward each of these substrates. In contrast, the monomorphic NAT isozyme of both acetylator genotypes showed apparent Vmax levels of NAT activity that did not vary with acetylator genotype. Furthermore, the monomorphic NAT isozyme did not show acetylator genotype-related variations in apparent Km toward the arylamine carcinogens AF and ABP, although differences were noted for PABA and AcCoA. These results suggest that the acetylator genotype-dependent expression of AcCoA-dependent NAT activity in hamster hepatic cytosol toward arylamines is primarily accountable by structural variants (allozymes) of polymorphic NAT under the genetic regulation of the acetylator gene locus. The acetylator genotype-independent expression of isoniazid NAT activity is attributable to a common monomorphic NAT isozyme in both acetylator genotypes. PMID- 2568904 TI - Nasal absorption and metabolism of progesterone and 17 beta-estradiol in the rat. AB - The utility of the nasal route for administration of progesterone and 17 beta estradiol has been studied in rats. The results indicate that both steroids are rapidly absorbed from the nasal cavity. The bioavailability of 14C-radiolabeled progesterone administered nasally was found to be 100% of that of an iv dose. Also, the areas under the curve for the iv and nasal routes of administration were in each case directly proportional to the dose within the range studied (50 150 micrograms/rat). The bioavailability of progesterone after intraduodenal (id) administration was only 1.2% of that following an iv dose. Nasal administration of 17 beta-estradiol resulted in significantly higher blood levels than those observed following id administration. The nasal bioavailability of 17 beta estradiol, calculated from the ratio of areas under the curve following nasal and iv administration, was 50%, 71%, and 84% for doses of 5, 10, and 20 micrograms per rat, respectively, compared with 2-5% via the id route for the same dose. The ratios of estrone to estradiol obtained after nasal administration of the different doses were lower than those obtained after id administration but higher than those obtained after iv administration, indicating that 17 beta-estradiol is oxidized to estrone in the nasal cavity and that the extent of oxidation is lower than that observed after id administration. The data also indicate that for both the iv and nasal routes of administration, estrone and estradiol constitute together 78% of the total unconjugated estrogens, whereas for the id route, these two steroids constitute 10-20% of the total unconjugated estrogens.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568906 TI - Characteristics of warfarin hydroxylation catalyzed by human liver microsomes. AB - The oxidative biotransformation of (R)- and (S)-warfarin was studied in human liver microsomes to determine whether an in vitro model could be established that would correspond to the in vivo profile that is generally observed. The quantitative pattern of oxidized products obtained from warfarin in vitro changed dramatically as a function of substrate concentration. Apparent Km values for the formation of 4', 6, 7, and 8-hydroxywarfarin indicated the presence of two easily distinguishable subsets of human liver cytochrome P-450; a high affinity subset (Km 3-15 microM) and a low affinity subset of isozymes (Km greater than 200 microM). The high affinity subset is primarily responsible for the metabolic profile of the biologically more potent (S)-enantiomer in vivo, whereas the low affinity subset is largely responsible for metabolism of the (R)-enantiomer. Apparent Vmax values alone did not reflect the relative in vivo formation clearances of the phenolic metabolites from either antipode, because the low affinity-high capacity component masked the metabolic profile of the (S) enantiomer. However, the rank order of intrinsic clearance, Vmax/Km, for each metabolite was in good agreement with regio- and stereoselective metabolism in vivo. This investigation highlights the need for rigorous kinetic characterization of an in vitro model before reasonable correlation can be expected with in vivo data. PMID- 2568905 TI - Human intestinal phenol sulfotransferase: assay conditions, activity levels and partial purification of the thermolabile form. AB - Phenol sulfotransferase (PST) catalyzes the sulfate conjugation of catecholamines and phenolic and catechol drugs. The small intestine is an important drug metabolizing organ. Other human tissues including brain, liver, and platelet contain at least two forms of PST. A thermolabile (TL) form catalyzes the sulfate conjugation of dopamine and other monoamines whereas a thermostable (TS) form of the enzyme catalyzes the sulfate conjugation of "simple" phenols such as p nitrophenol. In this study we found that human jejunal mucosa also contains at least two forms of PST with substrate specificities, sensitivities to inhibitors, and thermal stabilities similar to those of TL and TS PST in other human tissues. Optimal conditions were determined for the assay of TL and TS PST activities in jejunal mucosal preparations. Apparent Km values of TL PST for dopamine and of TS PST for p-nitrophenol were 10 microM and 0.92 microM, respectively. Jejunal mucosal TL PST, like TL PST in other human tissues, also catalyzed the sulfation of p-nitrophenol, but with a very high apparent Km of 1100 microM. PST activities were then assayed in 64 samples of jejunum obtained surgically, and average activities for TL and TS PST were 102 +/- 4.6 and 47.4 +/- 2.9 units per milligram of protein, (mean +/- SE), with 40 microM dopamine and 2 microM p nitrophenol as substrates, respectively. Since jejunal mucosal TL PST had a much higher specific activity than that present in other human tissues, TL PST was partially purified from jejunal mucosa by ion exchange and Affi-Gel Blue chromatography. The substrate specificity, inhibitor sensitivity, and physical properties of partially purified jejunal TL PST were similar to those of TL PST in supernatants of jejunal mucosal homogenates. True Km values of the partially purified enzyme for dopamine and p-nitrophenol were 6.7 microM and 1400 microM, respectively. Human jejunal TL PST had an apparent molecular mass of 69 kDa with 35.5-kDa monomers. In summary, human jejunal mucosa contains forms of PST similar to those found in other human tissues, and human jejunal mucosa is a good source for the purification of TL PST. PMID- 2568907 TI - Some factors affecting phencyclidine biotransformation by human liver and placenta. AB - Samples of human liver and placenta microsomes were analyzed for their in vitro hydroxylation capabilities using phencyclidine, [PCP, 1-(1 phenylcyclohexyl)piperidine] as substrate. Microsomes were prepared from full term placentas (cesarean deliveries under epidural anesthesia) and from histologically normal liver specimens (staging laparotomies for Hodgkin's disease). Three different hydroxylated PCP metabolites were assayed including 1 (1-phenyl-3-hydroxycyclohexyl)piperidine (3-OH-cyclo-PCP), 1-(1-phenyl-4 hydroxycyclohexyl)piperidine (3-OH-cyclo-PCP), 1-(1-phenyl-4 hydroxycyclohexyl)piperidine (4-OH-cyclo-PCP), and 1-(1-phenylcyclohexyl)-4 hydroxypiperidine (4-OH-pip-PCP). The mean amounts of in vitro microsomal hydroxylation of PCP at the three different positions of the PCP ring varied considerably between individual samples of both liver and placenta. The placenta hydroxylated PCP but not as effectively as liver. Evidence for independent hydroxylation of PCP to 3-OH-cyclo-PCP was comparable to 4-OH-cyclo-PCP and 4-OH pip-PCP. The formation of 3-OH-cyclo-PCP by the liver was enhanced in tobacco smokers. The formation of 4-OH-cyclo-PCP by the liver was negatively correlated with the stage of Hodgkin's disease even though the liver was free of disease in 11 of 12 subjects. PMID- 2568908 TI - Identification of etretinate metabolites in human bile. AB - The metabolites of etretinate (Tegison) were investigated in bile obtained from two patients with biliary T-tubes. Bile samples were collected for 5 days after administration of a single, oral 100-mg dose of 14C-labeled etretinate. Radioactivity measurements indicated that the two patients excreted 9.6% and 8.0% of the dose in the 5-day bile. The etretinate metabolites in the bile were present mainly as beta-glucuronidase-labile conjugates. HPLC analyses of the beta glucuronidase-treated bile samples showed no measurable concentrations (less than 10 ng/ml) of etretinate or the 13-cis acid, isoacitretin. Acitretin, the free acid of etretinate, was present and accounted for about 0.9% of the biliary radioactivity. The major portion of the radioactivity in the extracts of the beta glucuronidase-treated bile samples consisted of two metabolites with shortened side chains. One was identified as the 11, 12-dihydro-13, 14, 15, 20-tetranor phenolic derivative of acitretin, which was previously identified as a human urinary metabolite. The other metabolite was identified as the 11, 12, 13, 14 tetrahydro-15-nor phenolic derivative of acitretin, which has not been previously identified as a metabolite of etretinate. PMID- 2568910 TI - Stereochemical inversion of haloxyfop in the Fischer 344 rat. AB - The 2-aryloxypropionate haloxyfop is currently being evaluated for use as a herbicide. This compound is structurally similar to a group of 2-arylpropionates that have been shown previously to undergo stereochemical inversion in a variety of mammalian species. To support the data obtained from a number of toxicity/oncongenicity studies, in which racemic haloxyfop was employed, the stereochemical disposition of this compound was examined in the Fischer 344 rat. After administration of racemic haloxyfop (11 mg/kg, po) to male and female Fischer 344 rats, the day 1-10 urine samples were fortified with D4-haloxyfop (center ring label) and extracted, and the haloxyfop enantiomers were converted to diastereomeric derivatives [(S)-phenylethylamine] and analyzed by gas chromatography-mass spectrometry (GC/MS). Fecal samples for days 1-10 were fortified with D4-haloxyfop, extracted, and purified by reverse phase high pressure liquid chromatography prior to derivation and GC/MS analysis. In the female rat, 77.3% of the administered dose was recovered in the day 1-10 urine and feces as parent compound. The stereochemistry of the haloxyfop present in these samples was found to be nearly all (R)-enantiomer (greater than 98%). In the male rat, 52.2% of the dose was recovered in the day 1-10 urine and feces as haloxyfop. The stereochemistry of the parent compound present in these samples was similar to the results seen in the female rat [greater than 98% (R)]. These results show that (S)-haloxyfop undergoes rapid and nearly complete inversion to the (R)-enantiomer in the female Fischer 344 rat. The data also suggest a similar stereochemical inversion of haloxyfop in the male Fischer 344 rat. PMID- 2568909 TI - Identification of etretinate metabolites in human blood. AB - Five metabolites were isolated from the blood of psoriatic patients on chronic therapy with etretinate (Tegison). The two major metabolites were the all-trans acid (acitretin) and the 13-cis acid (isoacitretin), both of which had been previously identified as major metabolites of etretinate in human blood. The other three metabolites had not been previously reported in human blood. One metabolite corresponded to isoacitretin with a hydroxy group on an aromatic methyl group. The other two metabolites had acid side chains shortened by one carbon and a reduced 11, 12-double bond. One of the two metabolites retained the aromatic methoxy group, whereas the other was the phenolic analog. PMID- 2568911 TI - Isolation of 10 cyclosporine metabolites from human bile. AB - Ten metabolites of cyclosporine were isolate from the ethyl ether extract of bile from four liver transplant patients receiving cyclosporine. Two of the metabolites were unique and previously unidentified. Liquid-liquid partitioning into diethyl ether with subsequent defatting with n-hexane was used for the initial extraction from bile. Separation of the individual metabolites (A-J) was performed using a Sephadex LH-20 column and a gradient high performance liquid chromatographic method. The molecular weights of the isolated metabolites were determined by fast atom bombardment/mass spectrometry. Gas chromatography with mass spectrometric amino acid analysis was also used to identify the amino acid composition and the hydroxylation position of metabolites A, B, C, D, and G. Proton nuclear magnetic resonance spectra were utilized to distinguish the chemical shifts of N-CH3 singlets and NH doublets of metabolites A, B, C, and D. Metabolites A, E, F, H, I, and J were reported previously in human urine and animal bile. Metabolites C and D are dihydroxylated compounds which cannot be clearly described as previously isolated compounds. Metabolites B and G are novel metabolites with a mass fragment which corresponded to a loss of 131 Da from the protonated molecular ion (MH+) in the fast atom bombardment/mass spectrometry, suggesting that the double bond in amino acid 1 has been modified. Metabolites B and G were primarily isolated from the bile of one of the liver transplant patients which contained abnormally high concentrations of these two metabolites. The method described is an efficient procedure for isolating milligram quantities of the major metabolites with greater than 95% purity. PMID- 2568914 TI - Metabolism of bambuterol in rat liver microsomes: identification of hydroxylated and demethylated products by liquid chromatography mass spectrometry. AB - The oxidative metabolism of (R,S)-bambuterol in rat liver microsomes was studied. Metabolite fractions were analyzed by thermospray LC-MS. The use of an equimolar mixture of deuterium-labeled and unlabeled bambuterol facilitated the mass spectrometric identification of the metabolites. Six metabolites, formed via hydroxylation, demethylation, and hydrolytic reactions, were identified. The demethylated metabolites were found to be chemically unstable under physiological conditions. It is likely that the complex biotransformation of bambuterol into terbutaline is one factor contributing to the long duration of action of bambuterol. PMID- 2568913 TI - Disposition of the radioprotector ethiofos in the rhesus monkey. Influence of route of administration. AB - Plasma concentrations of ethiofos [S-2-(3-aminopropylamino)ethyl phosphorothioic acid, WR-2721] were compared following iv, ip, intraduodenal, and portal administration to the rhesus monkey. Plasma samples were analyzed for ethiofos, free WR-1065, [2-(3-aminopropylamino)ethanethiol], and total material convertible to WR-1065 (total WR-1065). In separate experiments, total radioactivity in plasma was compared following iv, ip, and intraduodenal administration of [14C]ethiofos; excretion of the radiolabel was measured in urine and in feces. Intraduodenal administration of unlabeled ethiofos rarely gave measurable levels of unchanged drug in plasma. In contrast, intraduodenal administration of [14C]ethiofos produced an average AUC for total radioactivity that was 62% of that for a 10-min iv infusion of [14C]ethiofos. Urinary excretion of radioactivity following iv and intraduodenal administration of [14C]ethiofos was 78.9 +/- 14.0% and 43.8 +/- 12.4%, respectively, whereas 1.9 +/- 0.5% and 9.7 +/- 6.3% was excreted in feces. After an ip dose of either labeled or unlabeled ethiofos, absorption of the dose was prolonged, but AUC values for total radioactivity or ethiofos and total WR-1065 were similar to those observed after the corresponding 10-min iv experiments. For either iv or portal routes, increases in ethiofos AUC values were observed for the same total dose when the infusion rate was increased from 1.25 to 15 mg/kg/min.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568915 TI - Metabolism of the antimycotic agent, croconazole, in rabbits. AB - The biotransformation of croconazole, a potent new antimycotic agent, was studied in the rabbit. Croconazole was excreted in the urine primarily as conjugates. Most of the radioactive metabolites in the urine could be extracted with organic solvent after hydrolysis with beta-glucuronidase. As many as 16 metabolites in the organic extracts were separated by TLC. Thirteen were identified by comparison of their mass and/or proton NMR spectra with those of synthetic samples. Aromatic ring hydroxylation of each benzene ring, O-dechlorobenzylation, and loss of the imidazole ring were found to occur. PMID- 2568912 TI - Quantitation of multiple pathways for the metabolism of nephrotoxic cysteine conjugates using selective inhibitors of L-alpha-hydroxy acid oxidase (L-amino acid oxidase) and cysteine conjugate beta-lyase. AB - In this study, we have established the selectivity of inhibitors for rat kidney cysteine conjugate beta-lyase and L-alpha-hydroxy acid oxidase (L-amino acid oxidase) and have used these inhibitors to explore the relative roles of these two enzymes in the metabolism of nephrotoxic cysteine conjugates by rat kidney homogenate. In addition, we have investigated the relationship between structure and the metabolism of toxic cysteine conjugates by purified rat kidney L-alpha hydroxy acid oxidase. With purified enzyme, S-(1,2,3,4,4-pentachlorobutadienyl)-L cysteine (PCBDC) was about four times more active than S(1,2-dichlorovinyl)-L cysteine (DCVC). Three alkyl conjugates were less active than DCVC. Purified L alpha-hydroxy acid oxidase was not inhibited by the beta-lyase inhibitor aminooxyacetic acid but was inactivated by 2-hydroxy-3-butynoate. PCBDC metabolism in rat kidney homogenate was inhibited 74% by aminooxyacetic acid and 42% by 2-hydroxy-3-butynoate, whereas DCVC metabolism was inhibited 77% by aminooxyacetic acid and 28% by 2-hydroxy-3-butynoate. However, only aminooxyacetic acid inhibited the binding of 35S label from [35S]DCVC. Based on these results we have reached three conclusions. First, L-alpha-hydroxy acid oxidase plays a significant role in the metabolism of some cysteine conjugates. Second, metabolism of DCVC by L-alpha-hydroxy acid oxidase does not contribute directly to covalent binding. Third, as much as 65% of DCVC may be metabolized to its corresponding alpha-keto acid. The results are discussed with regard to the nephrotoxicity of cysteine conjugates. PMID- 2568916 TI - Haloperidol reductase in human and guinea pig livers. AB - A GC assay method for haloperidol reductase was developed. Haloperidol reductase was present in guinea pig liver cytosol as well as in the microsomes, while in human liver only the cytosol exhibited the activity. The reductase activity was NADPH dependent for both species. Known substrates of ketone reductase such as menadione, daunorubicin, and ethacrynic acid strongly inhibited the human haloperidol reductase. The haloperidol reductase showed characteristics of ketone reductase. PMID- 2568917 TI - Evidence in humans for variant allozymes of the nondeficient sparteine/debrisoquine monooxygenase (P45OIID 1) in vitro. AB - Sparteine oxidation is part of a genetic polymorphism that affects the metabolism of many drugs and is under monogenic control. By examining the sparteine oxidation kinetics and the ratio of the dehydrogenated metabolites and through the use of the potent inhibitor, quinidine, two sites of metabolism were found for all 10 of the livers studied. The mean Km (N = 10) for the quinidine sensitive enzyme is 73 +/- 46 (SD) microM and the mean Vmax is 4.51 +/- 4.16 nmol/mg microsomal protein/30 min, indicating a large interindividual variation. Because the polymorphic defect is due to at least three variants of a mRNA splicing error with consequent lack of enzyme formation [Gonzalez et al.: Nature 331, 442 (1988)], the variation that we observed in Km is most likely due to variation of allozymes from extensive metabolizer alleles. The low affinity enzyme also demonstrates a large interindividual variation, is not competitively inhibited by quinidine, and produces a higher ratio of 5-dehydrosparteine to 2 dehydrosparteine than the high affinity enzyme. This low affinity enzyme must be part of a separate enzyme system from that controlling the sparteine/debrisoquine polymorphism because of its different characteristics and the 100% frequency with which it is found in the livers. The two dehydrosparteine metabolites are thought to be formed by the spontaneous breakdown of a primary metabolite. The different ratio of these two dehydrosparteines, which was found at low and high substrate concentrations, suggests that the reactions producing the primary metabolite are different between the quinidine-sensitive and -insensitive enzymes. PMID- 2568918 TI - Transport of acetaminophen conjugates in isolated rat hepatocytes. AB - The membrane permeabilities of acetaminophen glucuronide and sulfate produced through conjugative metabolism were examined in isolated rat hepatocytes. The glucuronide formed in hepatocytes was gradually released into the medium and its intracellular level decreased. Release of the sulfate formed in hepatocytes occurred more rapidly and its intracellular level remained almost constant. The permeability of acetaminophen was so rapid that it caused instantaneous equilibrium between hepatocytes and the medium. Its intracellular level thus decreased by conjugation reactions, but compensation for this decrease from the medium was soon made so that a constant intracellular level was resumed. The uptake of both preformed glucuronide and sulfate into hepatocytes indicated carrier-mediated transport. From these results, a pharmacokinetic model is proposed in which conjugative metabolism occurs in two consecutive steps: conjugative reactions of the parent compound taken up instantaneously into hepatocytes and membrane transport of conjugates into the medium. Changes in the amount of acetaminophen, its glucuronide, and its sulfate in the hepatocytes and medium as a function of time simulated according to the model closely agreed with those actually observed. Consequently, membrane permeability of the conjugates was concluded to be essential for conjugative metabolism. PMID- 2568920 TI - Stimulation of in vitro bioactivation of hydroquinone by phenol in bone marrow cells. PMID- 2568919 TI - Synthesis and hydrolytic behavior of the sulfate conjugate of 2'-hydroxy-3,4,4' trichlorocarbanilide. PMID- 2568921 TI - Carbonyl sulfide: a copper chelating metabolite of disulfiram. PMID- 2568922 TI - Khat chewing spread to the Somali community in Rome. AB - The habit of chewing Khat (Catha edulis) to experience its euphorizing and psychostimulant effects has prevailed for centuries among the inhabitants of the Horn of Africa and the Arabian peninsula. In recent years, air transport has removed the major obstacle to the diffusion of Khat, its perishableness. Khat is now air-freighted to Europe and we were able to buy some in Rome. We report the results of interviews with 20 members of the Somali community in Rome, who had continued their habit of chewing Khat whilst abroad. They asserted that they gathered together whenever possible, but preferably at weekends, to chew moderate quantities of Khat (one bundle, about 400 g). The customary habit of drinking tea or other soft drinks containing methylxantines during Khat sessions was maintained and very few subjects admitted drinking alcohol. In this small study sample, Khat chewing still seems to be a social event, as it is in Somalia. PMID- 2568923 TI - [Benzodiazepines at the general hospital. An example of their use at the Perigueux Hospital Center]. AB - The authors study prevalence and characteristics of BZD use in a general hospital (Perigueux, France). They try to point out the particularities of user's population (age, sex, psychiatric past, pathology at admission), the characteristics of prescription (nature, indication, dose, time of use), the place of BZD among the other anxiolytic and hypnosedative drugs employed. They come to the conclusion that strong prevalence of BZD use is not related to abusive prescription during hospitalization but to the lack of criticism concerning former prescriptions. PMID- 2568924 TI - Effects of sex steroids on the response of cultured rat pituitary cells to growth hormone-releasing hormone and somatostatin. AB - In the male rat, GH secretion is characterized by high amplitude pulses that appear at regular intervals of 3-4 h, with low basal levels between such pulses. In the female, the pulses are irregular and more frequent, with lower amplitudes, while basal secretion is higher. The present study was designed to exclude the indirect effects of sex steroids on the pituitary, enabling investigation of the direct effects of sex steroids on the pituitary. Rats were gonadectomized at 22 days of age, and 12 days later their anterior pituitaries were trypsinized for cell dispersion. Testosterone (T) or 17 beta-estradiol (E2), 5 nM, was added to the medium for 6 days, and subsequently, GRF or somatostatin was added for 4 h. In a perifusion system, the male-derived cell response to GRF was augmented after pretreatment with T, but not with E2. The female-derived cell response to GRF was augmented by E2, but not by T. T increased the sensitivity of the cells to GRF from 3.0-0.03 nM and increased the maximal potency of GH secretion 3-fold. E2 had no significant effect on the sensitivity, but lowered the potency. Somatostatin (1 nM) inhibited GH secretion by 44% in T-treated cells. In E2-treated cells, somatostatin was ineffective. GRF increased the total amount of GH (medium plus cells) in both T- and E2-treated cells, but not in control pituitary cells. It is suggested that T has direct effects on the male somatotroph. By increasing the pituitary cell responses to GRF and somatostatin, T contributes to the high amplitude peak/low baseline pattern of the male. By decreasing the pituitary cell responses to GRF and somatostatin, E2 contributes to the low amplitude peak/high baseline pattern of the female. PMID- 2568926 TI - Evidence for multiple protein constituents of the somatostatin receptor in pituitary tumor cells: affinity cross-linking and molecular characterization. AB - Previous studies have shown that somatostatin receptors on AtT-20 and GH3 pituitary tumor cells show relative preference for binding somatostatin-28 (S-28) and somatostatin-14 (S-14), respectively. Here we have attempted to determine whether this selectivity can be explained by molecular heterogeneity of the receptor. Cells were incubated with [125I-Tyr11]S-14, [125I-Leu8-D-Trp22,Tyr25]S 28, and [125I-Tyr3]SMS, and the bound ligand was chemically cross-linked with bis [2-succinimido-oxycarbonyloxy)ethyl]sulfone, disuccinimidyl suberate, or dithiobis (succinimidyl propionate). The solubilized cross-linked material was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by autoradiography. [125I-Tyr11]S-14 labeled three specific receptor proteins of 57K, 42K, and 27K mol wt in AtT-20 cells. The relative proportions of the protein bands were unaltered by the use of whole cells or cell membranes or by the inclusion of dithiothreitol or antiproteolytic agents. With both [125I-Tyr11]S-14 and [125I-LTT]S-28, the 57K protein constituted the major labeled component, representing 70-75% of the total cross-linked proteins. Labeling of the three protein species by [125I-Tyr11]S-14 and [125I-LTT]S-28 was inhibited by both S-14 and S-28 in a dose-dependent manner. S-28 was 10-20 times more potent than S-14 for inhibiting the labeling by both ligands of the principal receptor species of 57K. By contrast, when a radioiodinated derivative of the octapeptide analog octreotide ([125I-Tyr3]SMS) was used as ligand, the 27K protein was preferentially labeled, whereas the 57K and 42K bands were detected only as minor components. Labeling of GH3 cells with [125I-Tyr11]S-14 and [125I-LTT]S-28 revealed three cross-linked proteins of 57K, 42K, and 27K mol wt similar to those observed in AtT-20 cells. However, in this cell line the 27K protein, not the 57K species, was the dominant component identified with these two ligands, comprising 40-50% of the total cross-linked proteins. These results suggest that there are three somatostatin receptor proteins of 57K, 42K, and 27K in pituitary cells. In AtT-20 cells, the 57K protein constitutes the major receptor protein labeled by [125I-Tyr11]S-14 and [125I-LTT]S-28, whereas the 27K protein is the major species labeled by [125I-Tyr3]SMS. The 27K, not the 57K, moiety is the principal receptor form in GH3 cells. Such ligand- and tissue-selective binding by the somatostatin receptor provides strong evidence for receptor molecular heterogeneity. PMID- 2568925 TI - Tyrosine hydroxylase in the stalk-median eminence and posterior pituitary is inactivated only during the plateau phase of the preovulatory prolactin surge. AB - This study examined changes in the activity of tyrosine hydroxylase (TH) in the stalk-median eminence (SME) and posterior pituitary (PP) during the preovulatory PRL surge. Immature female rats were injected with PMSG on day 28. Blood PRL levels were low on the morning of day 30, rose to a peak from 1400-1600 h, remained at a lower plateau from 1800-2400 h, and declined to basal levels on the morning of day 31. SME, PP, and striatum were removed from PMSG-treated rats at selected times during the periovulatory period and from age-matched control rats. TH activity was determined in tissue homogenates by a coupled hydroxylation decarboxylation assay. Apparent Km and maximum velocity values with respect to 6 methyl tetrahydropterine were estimated from substrate saturation curves. The kinetic parameters for TH in either the SME or the PP of control rats were similar at 1100 and 1800 h on day 30. However, the apparent Km in both tissues was significantly lower than that in the striatum. The affinity of TH in the SME and PP was unchanged before and during the peak phase of the PRL surge, reduced significantly during the late plateau, and returned to presurge levels in the morning of day 31. TH activity in the striatum was similar at all times examined. To determine the state of activation of the enzyme, tissue homogenates were preincubated with cAMP, ATP, and magnesium. TH activity in the SME during the peak phase was unchanged by cAMP, and that in the PP was modestly increased. The relatively inactive enzyme in both tissues during the plateau phase was markedly activated by a cAMP-dependent mechanism. The low affinity of striatal TH was greatly increased by cAMP at both times. These data suggest that TH in the SME and PP exists in an activated state most of the time and is transiently inactivated during the plateau phase of the PRL surge. In contrast, TH in the striatum is relatively inactive in the basal state and is not affected by hormonal changes induced by PMSG. We conclude that the peak PRL surge occurs in spite of active dopamine (DA) neurons, suggesting that it is generated by a nondopaminergic mechanism. Decreased TH activity in DA neurons in the SME and PP may prolong the PRL surge during the plateau phase, whereas increased DA activity coincides with the termination of the surge. PMID- 2568927 TI - Anticonvulsant and antiarrhythmic actions of the beta blocking agent timolol. AB - Cats were anesthetized with alpha-chloralose and pentylenetetrazol, 10 and/or 20 mg were administered intracerebroventricularly (i.c.v.) to elicit epileptiform activity, including both interictal and ictal discharges. Timolol, 10, 100, 500 micrograms/kg i.c.v. and 1, 5, 10 and/or 20 mg/kg i.v., was administered at 5 min intervals to determine whether it suppressed the epileptiform activity. Mean arterial blood pressure and heart rate increased after the administration of pentylenetetrazol; these increases were associated with the development of epileptiform activity and cardiac arrhythmias. All doses of timolol caused a decrease in the blood pressure and heart rate elevated by pentylenetetrazol and suppressed the epileptiform activity. Similar findings were obtained in cats that received the same doses of timolol administered at different time intervals. The data indicate that the central administration of timolol reverses the epileptiform activity of pentylenetetrazol in the brain and suppresses the associated increases in blood pressure, heart rate and cardiac arrhythmias. PMID- 2568928 TI - Equilibrium, kinetic and photoaffinity labeling studies of daunomycin binding to P-glycoprotein-containing membranes of multidrug-resistant Chinese hamster ovary cells. AB - The binding of daunomycin and its Bolton-Hunter derivative iodomycin to plasma membranes isolated from multidrug-resistant Chinese hamster ovary cells (CHO B30) and their drug-sensitive parents (B1) was investigated. The thermodynamics and kinetics of equilibrium binding monitored by fluorescence titrations and temperature-jump relaxation spectrometry were compared with the specificity of covalent photolabeling with [3H]daunomycin and [125I]iodomycin. The facts that the uptake of anthracycline from aqueous solution into the CHO membranes was not accompanied by any substantial increase of fluorescence anisotropy nor by any spectral shift of the fluorescence emission spectrum and that the partition ratio into the membrane was 20-30-fold higher when compared to a lecithin bilayer, provided evidence that the non-covalent drug binding sites are constituted by polar protein domains without any substantial contribution from the surrounding lipids. Photoaffinity labeling with nanomolar concentrations of anthracycline and equilibrium binding curves independently showed that a 150-170-kDa plasma membrane glycoprotein (P-glycoprotein), whose overexpression is the major difference between B1 and B30 membranes, provides the binding sites of highest affinity for daunomycin and iodomycin (K approximately equal to 4 x 10(7) M-1). Comparison of photolabeling and equilibrium data suggested that the same binding sites on P-glycoprotein were most probably being monitored. The photolabeling of P-glycoprotein by iodomycin was inhibited in a dose-dependent manner by other compounds to which multi-drug-resistant cells are either resistant or collaterally sensitive with the following orders of effectiveness: vinblastine greater than verapamil greater than nitrendipine greater than daunomycin much greater than colchicine. Temperature-jump experiments covering the time range of 1 microseconds to 1 s revealed a single concentration-dependent relaxation time of 10-30 microseconds. The association of daunomycin with its binding sites in the membranes was found to be a diffusion-controlled process with kon rates of 2 4 X 10(9) M-1 s-1. Therefore, the selectivity of drug binding was entirely reflected in the dissociation rates. PMID- 2568929 TI - Pharmacokinetics of famotidine in elderly patients with and without renal insufficiency and in healthy young volunteers. AB - The pharmacokinetics of the H2-receptor antagonist famotidine, after oral administration of a 20 mg tablet, has been studied in 10 elderly patients with normal renal function (CLCR greater than or equal to 59 ml.min-1, Mean = 80 ml.min-1), 5 elderly patients with renal insufficiency (CLCR less than or equal to 38 ml.min-1, Mean = 15 ml.min-1), and 6 healthy young volunteers. Elimination half-life in the elderly patients with renal insufficiency was significantly prolonged compared to the elderly patients with normal renal function and the young volunteers. The correlation coefficient between creatinine clearance and the elimination rate constant of famotidine was 0.672. Mean urinary recovery of unchanged drug up to 24 h in the young volunteers was 44%. The mean renal clearance of famotidine in the young volunteers (270 ml.min-1) was substantially greater than the creatinine clearance, 128 ml.min-1, which suggests the possibility of tubular secretion of famotidine. PMID- 2568930 TI - Isolation and characterization of a cDNA encoding a putative cytokine which is induced by stimulation via the CD2 structure on human T lymphocytes. AB - To define activation-specific sequences in human T lymphocytes, a cDNA library was constructed by subtractive hybridization using resting and stimulated peripheral blood lymphocyte pairs. Stimulation of peripheral blood lymphocytes was achieved by triggering with mitogenic anti-CD2 (T11) monoclonal antibodies. Differential library screening with cDNA probes derived from stimulated vs. resting peripheral blood lymphocytes led to identification of a novel sequence, termed HC21. The predicted primary and secondary structure of HC21 deduced from the translated nucleotide sequence suggest that the gene encodes a secreted protein of 92 amino acids including a 23-residue leader sequence. Northern blot analysis demonstrates that HC21 mRNA is induced in peripheral blood T lymphocytes and interleukin 2-dependent T cell clones within 10 min following stimulation via CD2, while steady-state RNA expression is maximal by 2 h. Although the kinetics of HC21 expression are similar after stimulation via the antigen/major histocompatibility complex receptor (CD3-Ti), CD3-Ti triggering leads to less accumulation of HC21 RNA than CD2 triggering. In contrast, recombinant interleukin 2 does not induce detectable HC21 RNA expression in T cell clones. Transient expression of the HC21 cDNA in COS cells results in a readily detectable protein band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of radiolabeled cell supernatants consistent with HC21 encoding a secreted product. Protein sequence analysis reveals a striking homology (up to 76% identity at amino acid level) with other members of a recently described lymphokine family whose functions are yet to be defined. PMID- 2568932 TI - Effect of cyclosporin A on T cell immunity. II. Defective thymic education of CD4 T helper cell function in cyclosporin A-treated mice. AB - Cyclosporin A (CsA) is an immunosuppressive agent that is widely used in transplantation. Recent animal studies indicate that CsA can affect the development of immunity so that autoreactive T lymphocytes are generated. In this study, mice were treated with CsA prior to irradiation and transplantation of syngeneic bone marrow to determine whether CsA pretreatment would affect the ability of the bone marrow recipients to develop normal T cell function. Our results indicate that (a) thymuses of CsA-treated mice do not contain single positive thymocytes (i.e. L3T4+Ly-2- or L3T4-Ly-2+) during i.p. treatment with 15 mg/kg/day of CsA; (b) both populations of single-positive thymocytes reappear within 2 weeks of termination of CsA and (c) irradiation and bone marrow reconstitution of these CsA-treated mice results in reconstitution of normal numbers of L3T4+ and Ly-2+ cells, but the L3T4+ T cells to not provide T helper function, as determined by interleukin 2 production and cytotoxic T lymphocytes generation. These findings indicate that CsA can affect thymic microenvironment and may be important as a model for investigating intrathymic T cell maturation. Our results may also have clinical implications for T lymphocyte development in transplant patients receiving CsA. PMID- 2568931 TI - Interleukin 2 receptor expression and interleukin 2 production in exponentially growing T cells: major differences between in vivo and in vitro proliferating T lymphocytes. AB - In the present study we have assessed the growth requirements for in vivo proliferating mature T cells. For that purpose we have selected experimental approaches which allow the study of exponential growth in vivo of a major fraction of T cells, and make it possible to obtain large numbers of T cells in cycle. Two types of growing T cell populations were used: peripheral T lymphocytes, proliferating exponentially after transfer into syngeneic athymic nude mice, and activated T cells in lymph nodes of normal mice draining the site of oxazolone administration. The results obtained show that mature T cell growth in vivo is not accompanied by expression of high-affinity interleukin 2 (IL2) receptor in the majority of activated cells, is not abrogated by in vivo administration of anti-IL2 receptor antibodies or enhanced by the in vivo injection of recombinant IL2, and that in vivo growing T cells do not produce detectable amounts of IL2, as evaluated functionally by limiting dilution assays or the presence of IL2 mRNA, detected by Northern blots or in situ hybridization. The presented data thus indicate that the rules known to apply to T cell activation and proliferation in vitro differ from those used by in vivo growing T lymphocytes, at least in the two systems studied. PMID- 2568933 TI - Effects of histamine H3-receptor ligands on various biochemical indices of histaminergic neuron activity in rat brain. AB - The interaction of the potent histamine H3-receptor ligands i.e. (R)alpha methylhistamine, an agonist, and thioperamide, an antagonist, with the three classes of cerebral histamine receptors was studied in vitro and in vivo. The histamine-induced stimulation of 3',5'-cyclic AMP accumulation in slices of guinea-pig hippocampus was not modified by thioperamide (up to 0.1 mM) and (R)alpha-methylhistamine stimulated cyclic AMP accumulation only at millimolar concentrations. Hence, both (R)alpha-methylhistamine and thioperamide were at least 100,000-fold more potent at H3- than at H1- or H2-receptors in brain. In vivo, the turnover of histamine in rat cerebral cortex, as determined from its depletion elicited by alpha-fluoromethylhistidine in a synaptosomal fraction was not modified by mepyramine and zolantidine but was markedly enhanced by thioperamide at a low dose (ED50 = 2 mg/kg). Thioperamide also elicited a long lasting decrease in synaptosomal histamine and increase in radioimmunoassayable N tau-methylhistamine. In contrast, (R)alpha-methylhistamine markedly reduced cortical [3H]histamine synthesis (ED50 = 5 mg/kg). This long-lasting action was accompanied by an increase in synaptosomal histamine and a decrease in N tau methylhistamine levels. These changes were compared with those in plasma drug levels. Hence the two H3-receptor ligands appear to modify the activity of cerebral histamine neurons markedly and in a long-lasting and opposite manner. PMID- 2568934 TI - Bunazosin, an alpha 1-adrenoceptor blocker, differentially releases co transmitters in dog mesenteric vessels. AB - The effects of bunazosin on the electrical and mechanical responses of smooth muscle cells elicited by exogenously applied noradrenaline (NA) and by perivascular nerve stimulation were studied in the isolated mesenteric artery and vein of the dog. NA (above 10(-7) M in the artery and above 3 X 10(-8) M in the vein) depolarized the membrane. Perivascular nerve stimulation evoked an excitatory junction potential (e.j.p.) and slow depolarization in both vessels. Bunazosin and prazosin inhibited the NA-induced depolarization and slow depolarization in the artery but not in the vein. The NA actions in the vein were inhibited by yohimbine. Bunazosin (above 10(-6) M) increased the amplitude of the e.j.p. but decreased the outflow of NA during nerve stimulation. The amplitude and conduction velocity of the compound action potential of perivascular nerves were inhibited by higher concentrations of bunazosin (above 10(-5) M). The results provide evidence that bunazosin has selective inhibitory actions at alpha 1-adrenoceptors. This drug exerted differential effects on the release of co transmitters which generate the e.j.p. and the slow depolarization, as bunazosin increased the former and decreased the latter. This suggests that e.j.p. is generated by a substance other than NA. PMID- 2568935 TI - Characterization of the beta 1-adrenoceptor stimulatory effects of the partial beta 1-agonists acebutolol, xamoterol, H142/08 and H201/70. AB - The beta 1-adrenoceptor stimulatory effects of the partial beta 1-agonists acebutolol, xamoterol, H142/08 and H201/70 were investigated in the isolated right atrium (frequency response) of the rat. All the partial agonists studied induced a concentration-dependent increase in atrial rate. This effect was antagonized by the beta 1-selective blocker pafenolol. The concentrations of H142/08 and H201/70 needed to produce a half maximal response (-log EC50: pD2) were significantly greater than those required to occupy half the receptor population (-log equilibrium dissociation constant: pKB). These compounds required a fractional receptor occupancy of 80-90% to produce half the maximal stimulatory effect while the corresponding receptor occupancy for the other partial agonists studied was about 20%. The maximal stimulatory effect (intrinsic activity) generated by the compounds in the right atrium was (mean +/- S.D.): xamoterol 60 +/- 11%, H142/08 30 +/- 9%, H201/70 18 +/- 3% and acebutolol 17 +/- 8%. In addition, the stimulatory potency of the partial agonists was calculated as the efficacy (e) of the compounds relative to that of isoprenaline. The relative efficacy, expressed as -log[e(partial agonist/e(isoprenaline)] was: xamoterol 2.2 +/- 0.4, acebutolol 3.0 +/- 0.2, H142/08 3.4 +/- 0.2 and H201/70 3.6 +/- 0.2. It is concluded that partial beta 1-agonists have different relationships between their stimulatory effect and fractional receptor occupancy. There was a poor correlation between the intrinsic activity and relative efficacy of partial beta 1-agonists in the right atrium. PMID- 2568936 TI - [3H]SCH 23390 labels dopamine D-1 receptor sites in the rat kidney. AB - The binding of [3H]SCH 23390, a selective dopamine D-1 receptor radioligand, to rat kidney cortical membranes was studied. Scatchard analysis revealed a single class of binding sites. Of dopamine, noradrenaline and serotonin, dopamine was the most potent of these to displace [3H]SCH 23390 binding. The selective D-1 ligands SCH 23390 and SK & F 38393 were more potent to displace [3H]SCH 23390 than the selective D-2 ligands S-sulpiride and LY 171555, thus indicating that [3H]SCH 23390 binds predominantly to dopamine D-1 receptor sites. PMID- 2568937 TI - Amiloride interacts with [3H]idazoxan and [3H]rauwolscine binding sites in rabbit urethra. AB - Amiloride and its analogues (N-ethylisopropylamiloride and banzamil) interact more potently with [3H]idazoxan binding sites (nM range) than with [3H]rauwolscine binding sites (microM range) in the rabbit urethra. The binding of both radioligands was competitivelly inhibited by amiloride, with increased KD values and no change in their binding capacity. Interestingly, amiloride was a potent inhibitor at [3H]idazoxan binding sites in rabbit urethral smooth muscle at concentrations far below those required to inhibit the Na+/H+ exchanger or electrogenic pump. PMID- 2568938 TI - Adrenergic and purinergic neurotransmission in arterial resistance vessels of the cat intestinal circulation. AB - The contribution of adrenoceptors and purine receptors in mediating neurogenic vasoconstriction was investigated in the autoperfused intestinal circulation of anaesthetised cats treated with atropine and propranolol. Prazosin (0.5 mg/kg) and yohimbine (1.5 mg/kg) reduced but did not abolish the vasoconstrictor responses to stimulation of the efferent sympathetic nerves. The inhibitory actions of the two antagonists were additive but even after alpha 1- and alpha 2 adrenoceptor blockade nerve stimulation still elicited a residual, frequency related vasoconstriction. The initial, rapid, phase of this response was completely abolished after desensitisation of P2x-purinoceptors with a high dose (1.5 mg i.a.) of alpha,beta-methylene ATP. In the absence of alpha-adrenoceptor antagonists, alpha,beta-methylene ATP reduced neurogenic vasoconstriction particularly at low frequency (1 Hz) nerve stimulation, but also caused a short lasting decrease in noradrenaline and methoxamine responses which indicates that the drug may have some non-specific effects on arterial smooth muscle. The results suggest that neurotransmission in arterial resistance vessels of the cat intestinal circulation is predominantly under adrenergic control mediated by postsynaptic alpha 1- and alpha 2-adrenoceptors, with a possible purine involvement in the initial rapid response of the blood vessels, particularly to low frequency nerve stimulation. PMID- 2568939 TI - Effects of botulinum A toxin on presynaptic modulation of evoked transmitter release. AB - A possible influence of botulinum A toxin on the modulation of evoked neurotransmitter release was investigated in hippocampus tissue. Rabbit hippocampal slices prelabelled with [3H]noradrenaline ([3H]NA), [3H]5 hydroxytryptamine ([3H]5-HT) or [3H]choline were superfused with physiological medium and were stimulated electrically during superfusion. The evoked release of [3H]NA, [3H]5-HT and [3H]acetylcholine [( 3H]ACh) was inhibited by botulinum A toxin in a concentration- and time-dependent manner. Neither the inhibition of release of [3H]NA and [3H]5-HT by the alpha 2-adrenoceptor agonist clonidine nor facilitation of release in the presence of alpha 2-antagonists were influenced by pretreatment of the tissue with botulinum toxin. The toxin caused no [32P]ADP ribosylation of synaptosomal proteins of hippocampus. The facilitation of the stimulation-induced [3H]NA and [3H]5-HT release by the specific protein kinase C (PKC) activator 4 beta-phorbol-12,13-dibutyrate (PDB) was significantly diminished by botulinum A toxin. These results show that the evoked transmitter release is inhibited by botulinum A toxin by a mechanism which does not involve ADP ribosylation or an interaction with the alpha 2-adrenoceptor mechanism. PMID- 2568940 TI - Stimulation of peripheral cholinergic nerves by glutamate indicates a new peripheral glutamate receptor. AB - The bronchial smooth muscle of the rat was examined for contractile responses to excitatory amino acids. The nerve-mediated contraction induced by electrical field stimulation was enhanced by exogenous L-glutamate (L-Glu). The apparent affinity (ED50) of L-Glu was 3.5 +/- 0.1 mM. Both tetrodotoxin and hemicholinium 3 completely abolished the electrical field-induced contraction and therefore the potentiation by L-Glu, which indicates that L-Glu has a prejunctional effect. Concentrations of L-Glu higher than 22 mM inhibited the electrical field-induced contractions and enhanced the tonus of the smooth muscle by postjunctional stimulation. The ED50 of exogenous ACh was not altered by L-Glu. High concentrations (62 mM) of L-Glu increased the intrinsic activity (alpha) of ACh, indicating a postjunctional potentiation of ACh-induced contractions. L-Glu did not inhibit the activity of acetylcholinesterase, therefore the postjunctional potentiation was not due to ACh accumulation. Inhibition of the electrical field induced contraction was seen with high concentrations of D-Glu, L-aspartate (L Asp), L-alpha-amino adipate and ibotenate. Neither glutamate diethyl ester nor 2 amino-5-phosphonovalerate had any inhibitory effects on the L-Glu- and L-Asp induced alterations of the electrical field-stimulated contraction or on the L Glu-enhanced tonus of the bronchial smooth muscle. Kainate, N-methyl-D-aspartate, quisqualate and N-acetyl-aspartyl-glutamate had only minor transient potentiating effects on the electrical field-induced contraction. The results provide evidence for a L-Glu receptor in rat bronchi that has a different specificity for glutamate agonists and antagonists than the L-Glu receptor described in the CNS. The receptor seems to be located prejunctionally and enhances nerve-mediated responses and thereby stimulates the bronchial smooth muscle to contract. The possible involvement of this type of receptor in the 'Chinese restaurant syndrome' is discussed. PMID- 2568941 TI - Morphology of tyrosine hydroxylase-immunoreactive neurons in the human cerebral cortex. AB - In freshly fixed biopsies of human cerebral cortex obtained at surgery, immunocytochemical staining with antibodies against tyrosine hydroxylase (the rate limiting biosynthetic enzyme for catecholamines) revealed, in addition to a dense axonal plexus, a population of immunoreactive cell bodies. The neuronal nature of these cells was ascertained by: i) the presence of a rich rough endoplasmic reticulum in the cell body and of synapses on the cell body and dendrites, and ii) the demonstration of the lack of reactivity with the astroglial marker, glial fibrillary acidic protein, in the tyrosine hydroxylase immunoreactive cells. The tyrosine hydroxylase-immunoreactive neurons were found in all areas of cortex sampled, and were located almost exclusively in the infragranular layers. Most tyrosine hydroxylase-immunoreactive cells were bipolar and were vertically oriented, but a few had a multipolar or horizontal dendritic arbor. The dendrites of these cells were varicose and aspiny, and the axons were very thin. Tyrosine hydroxylase-immunoreactive neurons were reported to be present transiently in the developing mammalian cerebral cortex and only recently in cerebral cortex of mature mammalian brains. Internuncial neurons in the human cerebral cortex containing a catecholamine synthesizing enzyme would be significant, in particular considering that catecholamines are likely to be involved in some major mental disorders. PMID- 2568942 TI - Synchronized neuronal activities in neocortical explant cultures. AB - Intracellular recordings revealed that in neocortical explant cultures prepared on the day of birth and examined 3-6 weeks later, neurons mature and establish complex synaptic relationships that lead to spontaneous and triggered synchronous discharge. The spontaneous synchronous activity took several forms, including periodic generation of epileptiform depolarizing waves, prolonged periods of seizure-like discharge, and periodic, intense barrages of IPSPs. Synchronous depolarizations were associated with a marked increase in membrane conductance. Intracellular injection of currents of varying polarity and intensity affected their amplitudes and polarities without influencing the probability of their occurrence, indicating that the discharge reflected the synchronous activities of a neuronal population. This conclusion was confirmed with simultaneous recordings from pairs of neurons. Effects of the GABAa receptor antagonist, bicuculline, and the NMDA receptor antagonist, 2-aminophosphonvalerate (2APV), were used to assess the contributions of impairment of inhibition and enhancement of excitation to the initiation of synchronous discharge. The frequency with which spontaneous depolarizations were generated in normal medium was markedly reduced by 2APV. Moreover, seizure-like activity was induced by removing Mg++ from the medium, a condition that enhances conductance through NMDA receptor-coupled channels. This behavior was also attenuated by 2APV. Perfusion of bicuculline was potently epileptogenic. 2APV cut short the late, voltage-dependent phase of bicuculline induced paroxysmal depolarizations, indicating a role of NMDA receptors in generating this component of the wave. Epileptiform activities induced by withdrawal of Mg++ were greatly augmented by bicuculline, indicating that blockade of inhibition was not a prerequisite for seizure-like activity. This conclusion is supported by the finding that in many neurons in untreated cultures, paroxysmal generation of trains of IPSPs was the primary manifestation of spontaneous, synchronous population discharge. PMID- 2568943 TI - The role of inhibitory processes in the formation of functional properties of neurons in vibrissal projection zone of the cat somatosensory cortex. AB - The role of intracortical inhibitory processes in the formation of neuronal receptive fields in the vibrissal projection zone of the somatosensory cortex was studied. Iontophoretic application of picrotoxin and bicuculline blocks the inhibition and causes the loss of directional sensitivity in neurons. Activation of inhibition by distant glutamate application gives opposite results--neurons become direction sensitive. A dependence was found between spatial location of activated cells and the pattern of changes of their detector properties. Inhibitory processes caused by natural afferent stimulation lead to similar changes in the functional properties of neurons. PMID- 2568944 TI - Rodent and primate adrenal medullary cells in vitro: phenotypic plasticity in response to coculture with C6 glioma cells or NGF. AB - In order to maintain a chronic supply of growth factor for medulla cells in vitro, chromaffin cells from rat, African green monkeys and man were co-cultured with C6 glioma cells, which secrete growth factors that sustain sympathetic neurons in vitro. The response of chromaffin cells to coculture was compared to treatment of medullary cells with nerve growth factor (NGF) alone. Dispersed chromaffin cell preparations were obtained by a trypsin-collagenase procedure, and subjected to differential plating on collagen-coated surfaces. With both human and monkey tissue, non-chromaffin cells did attach to the culture plates and an enriched chromaffin cell population could be replated. Rat adrenal medulla cells survived very poorly in vitro and were not enriched in this procedure. Cultured human and monkey chromaffin cells survived as epithelial cells (50%) and showed neuritic outgrowth on 55 to 66% of the cells after eight days when treated with nerve growth factor (NGF). These cells showed strong catecholamine histofluorescence, tyrosine hydroxylase (TH) and dopamine beta hydroxylase (DBH) immunoreactivity. In contrast, only ten percent of adult rat chromaffin cells survived in culture, although NGF treatment rescued an additional 20% of the cells and induced neuritic outgrowth after one week in vitro. C6 glioma cells were treated with mitomycin C bromodeoxyuridine to inhibit mitosis and were plated with the various medulla cells in a one to one ratio. Both human and monkey chromaffin cells expressed extensive and enhanced neuritic arborization within eight days of co-culture, (64-82% respectively) and exhibited intimate contact with the glioma cells as seen at the ultrastructural level. Importantly, survival of adult rat adrenal medulla cells was enhanced to 50% or more with 40% of the cells extending neurites when co-cultured with glioma cells for seven days. Chromaffin cells from all three species reacted for TH, DBH and PNMT in co culture and were histo-fluorescent. The majority of these cells were also immunoreactive for serotonin and enkephalin, while only 37% of chromaffin cells indicated the presence of NPY. These data indicate that adrenal medulla can be maintained in vitro as the neuronal phenotype when co-cultured with growth factor producing cells and that this strategy may be useful for in vivo transplantation studies. PMID- 2568945 TI - GM1 ganglioside treatment promotes recovery of striatal dopamine concentrations in the mouse model of MPTP-induced parkinsonism. AB - GM1 ganglioside (GM1) has in the past been reported to promote regenerative sprouting and functional recovery in both central and peripheral nervous systems. The present experiments were performed in order to investigate whether GM1 might have any therapeutic effect on young mice who had been exposed to the Parkinson producing neurotoxin MPTP. GM1 caused moderate to dramatic increases in striatal dopamine levels, depending upon duration of exposure to GM1, in animals previously exposed to MPTP. Furthermore, the effects of GM1 on enhancing striatal dopamine levels were apparent when GM1 administration was delayed until 3 days after the last MPTP injection was given and these effects were not reversed when GM1 was withdrawn. Tyrosine hydroxylase (TH) immunohistochemistry of the striatum demonstrated increased numbers of TH-positive fibers and TH-positive terminal fields in GM1-treated animals as compared to animals that received only MPTP. TH immunohistochemistry of the substantia nigra revealed little or no loss of parts compacta neurons in the MPTP-treated mice. On the basis of these observations, GM1 appears to increase the dopamine content of the striatum by promoting or stimulating regenerative sprouting of dopaminergic terminals and perhaps collateral sprouting from remaining intact fibers in the MPTP model of Parkinsonism in the young mouse. We suggest that GM1 ganglioside may hold some promise as a potential adjunct in the treatment of Parkinson's Disease. PMID- 2568946 TI - Seizures evoked from area tempestas are subject to control by GABA and glutamate receptors in substantia nigra. AB - The functional relationship between the substantia nigra (SN) and the area tempestas (AT), an epileptogenic site in the deep prepiriform cortex, was investigated. Stimulation of GABA receptors in SN with muscimol, or blockade of nigral excitatory amino acid receptors with 2-aminophosphonoheptanoic acid (AP7), protected against convulsions evoked by the unilateral focal injection of bicuculline in the AT. The protective effects were obtained only with bilateral nigral injections; unilateral manipulations were without anticonvulsant effect. Bilateral intranigral morphine did not exert an anticonvulsant effect against convulsions evoked from the AT, although this treatment induced stereotyped sniffing and gnawing behavior similar to that evoked by intranigral muscimol and AP7. The data indicate that seizures evoked from the AT of one hemisphere are susceptible to suppression by inhibition in the SN, but only when the inhibition occurs in the SN of both hemispheres. This suggests that the seizure suppressant action evoked from the SN is exerted during the course of the propagation of the seizure, once it has spread bilaterally. PMID- 2568947 TI - Modulation by somatostatin of nerve-mediated activation of glycogenolysis in the perfused rat liver. AB - Perivascular nerve stimulation of rat livers perfused in situ with erythrocyte free Krebs-Henseleit buffer at constant pressure in a non-recirculating system resulted in an increase of glucose and lactate production and in a decrease of portal flow. Infusion of somatostatin in different concentrations (2 x 10(-7), 10(-8), 10(-9) mol.l-1) reduced the nerve-mediated activation of glucose release maximally to 66%. There was only a slight effect on the lactate output, the nerve mediated reduction of portal flow was unaltered. In controls, somatostatin alone had no effect on the metabolic and hemodynamic parameters. In order to differentiate between a presynaptic and postsynaptic mechanism, the noradrenaline overflow was calculated. The unaltered release of the neurotransmitter in the presence or absence of somatostatin excluded a presynaptic mechanism. To mimic the nerve effects on the carbohydrate metabolism and on the hemodynamics, noradrenaline (2 x 10(-7) mol.l-1) was infused instead of the nerve stimulation over a period of 5 min. Somatostatin did not change the endocrine effects of the catecholamine under these conditions. The nerve-dependent effect of somatostatin suggests that other neurotransmitters (e.g. VIP) or mediators (e.g. prostanoids) may be influenced by somatostatin. PMID- 2568948 TI - Binding of plasminogen to Escherichia coli adhesion proteins. AB - Immobilization of plasminogen via its lysine-binding sites is regarded as a prerequisite for its activation and function in fibrinolysis and pericellular proteolysis. In the present study, the interaction of plasminogen with fimbriae found on Escherichia coli strains causing invasive human infections was studied. Plasminogen displayed concentration-dependent and saturable binding to immobilized type 1 fimbriae and, several fold lower binding to P and S fimbriae. The binding to fimbriae was effectively inhibited by epsilon-aminocaproic acid indicating that it was mediated by the lysine-binding sites of plasminogen. Binding studies with mutated fimbriae and inhibition tests indicated that the interaction was not dependent on the lectin subunit of the fimbriae. These results indicate the existence of a novel type of host-microbe interaction which may be important in the invasion by bacteria of host tissues. PMID- 2568949 TI - Transformation of mouse mammary epithelial cells with the Ha-ras but not with the neu oncogene results in a gene dosage-dependent increase in transforming growth factor-alpha production. AB - An enhanced expression of transforming growth factor-alpha (TGF alpha) was demonstrated in two clones of NOG-8 mouse mammary epithelial cells, NOG-8 SR1 and NOG-8 SR2, that have been transformed by a v-Ha-ras oncogene. The amount of TGF alpha production in NOG-8 SR1 and NOG-8 SR2 cells was dependent on the level of p21ras expression in these clones, which directly correlated with their cloning efficiency in soft agar. There was also a decrease in the number of epidermal growth factor (EGF) receptors on the NOG-8 SR1 and NOG-8 SR2 cells that is proportional to the amount of TGF alpha secreted. These effects were specific for ras because neu-transformed NOG-8 cells grew in soft agar at a comparable level to NOG-8 SR2 cells yet did not show any increase in TGF alpha production or change in EGF receptor expression. PMID- 2568950 TI - Cloning of the pig aminopeptidase N gene. Identification of possible regulatory elements and the exon distribution in relation to the membrane-spanning region. AB - We have isolated four lambda-phages covering the complete pig aminopeptidase N/CD13 gene. The sequence of 2.85 kbp encompasses 1.18 kbp of the 5' upstream region and 1.67 kbp of the structural gene. In the promoter region we find a TATA box and potential binding sites for CTF-1/NF-1 and AP-2. By sequence comparisons we have found three domains showing similarity to promoter regions of the genes encoding human alpha 1-antitrypsin and human intestinal alkaline phosphatase. The gene sequence includes the first three exons and two introns. It shows that a single exon encodes the cytoplasmic tail, the membrane anchor and the junctional peptide. PMID- 2568951 TI - Identification of protein phosphatase 2A as the major tyrosine hydroxylase phosphatase in adrenal medulla and corpus striatum: evidence from the effects of okadaic acid. AB - (i) The major sites on bovine adrenal tyrosine hydroxylase (TH) phosphorylated by calmodulin-dependent multiprotein kinase (CaM-MPK) and cyclic AMP-dependent protein kinase were shown to be Ser-19 and Ser-40, respectively, while Ser-40 was also phosphorylated slowly by CaM-MPK. (ii) Type 2A and type 2C phosphatases accounted for approximately 90% and approximately 10% of TH phosphatase activity, respectively, in extracts of adrenal medulla and corpus striatum assayed at near physiological free Mg2+ (1 mM), while type 1 and type 2B phosphatases had negligible activity towards TH. (iii) Incubation of adrenal chromaffin cells with okadaic acid increased TH phosphorylation by 206% and activity by 77%, establishing that type 2A phosphatases play a major role in regulating TH in vivo. PMID- 2568953 TI - Consequences of bacterial attachment in the urinary tract. PMID- 2568954 TI - Interactions between enterocytes and enteropathogenic Escherichia coli. PMID- 2568952 TI - Synthesis and evaluation of optically pure [3H]-(+)-pentazocine, a highly potent and selective radioligand for sigma receptors. AB - Tritium-labeled (+)-pentazocine ([3H]-1b) of specific activity 26.6 Ci/mmol was synthesized in 3 steps starting with (+)-normetazocine (2) of defined optical purity. [3H]-1b has been characterized as a highly selective ligand for labeling of sigma receptors. Competition data revealed that [3H]-1b could be displaced from guinea pig brain membrane preparations with a number of commonly used sigma receptor ligands. [3H]-1b exhibited saturable, enantioselective binding with a Kd of 5.13 +/- 0.97 nM and a Bmax of 1146 +/- 122 fmol/mg protein. Phencyclidine (PCP) displaced [3H]-1b with low affinity while MK-801 was inactive, thus indicating insignificant activity at the PCP-binding site; apomorphine failed to displace [3H]-1b indicating lack of dopamine receptor cross-reactivity. Since the affinity of [3H]-1b is about 6 times that of the two commonly employed sigma ligands ((+)-3-[3H]PPP and [3H]DTG) and since it is more selective for sigma receptors than the benzomorphan [3H]SKF-10,047, it represents the first example of a highly selective benzomorphan based sigma receptor ligand. [3H]-1b should prove useful for further study of the structure and function of sigma receptors. PMID- 2568955 TI - Polymorphic human insulin-responsive glucose-transporter gene on chromosome 17p13. AB - Glucose uptake by heart, skeletal muscle, and adipose tissue is acutely regulated by insulin, which stimulates facilitative glucose transport, at least in part, by promoting the translocation of transporters from an intracellular pool to the plasma membrane. cDNAs encoding the major human insulin-responsive glucose transporter have been isolated and indicate that the insulin-responsive glucose transporter expressed by heart, skeletal muscle, and adipose tissue is a 509 amino acid protein having 65.3, 54.3, and 57.5% identity with the erythrocyte/HepG2, liver, and fetal muscle glucose transporters, respectively. The gene encoding the insulin-responsive glucose transporter (designated GLUT4) was mapped to the p11----p13 region of the short arm of human chromosome 17 by analyzing its segregation in a panel of reduced human-mouse somatic cell hybrids. In situ hybridization to prometaphase chromosomes indicated that GLUT4 was in band p13. A common two-allele restriction-fragment-length polymorphism (RFLP) was identified with Kpn I, and linkage of this RFLP to other polymorphic DNA markers in this region of chromosome 17 provides a set of probes that will be useful for examining the role of this gene in the pathogenesis of diabetes mellitus. PMID- 2568956 TI - Disease associations. Chance, artifact, or susceptibility genes? AB - Numerous genes that might contribute to the development of diabetes mellitus and/or its complications have been isolated and characterized. One approach to determining whether these "candidate" genes influence susceptibility to diabetes is to compare the frequency of a DNA marker(s) (restriction-fragment-length polymorphism) for each gene is appropriately matched groups of patients and control subjects. The identification of a DNA-marker association would suggest that genetic variation at this gene may increase or reduce the risk of developing diabetes. However, the absence of an association does not necessarily imply that this gene does not contribute to the development of diabetes. We discuss the genetic rationale of disease association studies and the importance of sample size and disease-marker allele frequencies in these studies. PMID- 2568957 TI - Tyrosine hydroxylase activity in sympathetic nervous system of rats with streptozocin-induced diabetes. AB - The activity of tyrosine hydroxylase (TOH), the rate-limiting enzyme in norepinephrine biosynthesis, was measured in selected sympathetic ganglia to develop a quantitative measure of sympathetic autonomic neuropathy in streptozocin-induced diabetic rats. Surprisingly, TOH activity was elevated twofold in diabetic prevertebral ganglia innervating the alimentary tract (i.e., superior mesenteric, celiac, and inferior mesenteric), which has terminal processes that develop neuroaxonal dystrophy in this model system. TOH activity of paravertebral ganglia (i.e., superior cervical and stellate) with nonalimentary targets was not increased in the same animals. Increased TOH activity in the prevertebral ganglia 1) developed within the 1st wk of diabetes and persisted for 10 mo, 2) did not represent a change in TOH affinity for d-1,6 methyl-5,6,7,8- tetrahydropterine cofactor, 3) was prevented by both nicotinamide pretreatment and early institution of insulin therapy, and 4) did not develop as a result of classical transsynaptic induction. Pair-feeding experiments confirmed that the most likely cause of increased TOH activity in this system was the marked hypertrophy and hyperplasia of the diabetic bowel resulting from compensatory hyperphagia. We conclude that TOH activity does not represent a suitable marker for sympathetic autonomic neuropathy in this experimental system. Rather, the increase appears to be an example of a selective increase in the synthesis of neurotransmitter enzymes, possibly in response to increased trophic support provided by the expanded target, i.e., the hypertrophic gut. The additional synthetic stress imposed on prevertebral neurons by the expansion of the innervation of the alimentary target coupled with the complex diabetic metabolic milieu may contribute to the development and selective distribution of dystrophic axonopathy to the innervation of the alimentary tract. PMID- 2568958 TI - Type II diabetes mellitus and polymorphism of insulin-receptor gene in Mexican Americans. AB - Resistance to insulin action is a well-established feature of type II (non insulin-dependent) diabetes and is believed by many to contribute to the etiology of this condition. We therefore characterized restriction-fragment-length polymorphisms of the insulin-receptor gene with the restriction enzyme Rsa 1 in 242 Mexican Americans and non-Hispanic Whites with type II diabetes and 202 age-, sex-, and ethnicity-matched control subjects who participated in a population based study in San Antonio. Alleles of 6.7 kilobases (kb) (A allele), 6.2 kb (B allele), and 3.4 kb (C allele) were identified. The C allele was observed in Mexican Americans only, where its frequency among nondiabetic control subjects was 17.7%. Diabetic Mexican Americans were twice as likely as control subjects to be homozygous for the C allele. The crude odds ratio for diabetes in CC homozygotes compared with the other two genotypes was 2.22, although this result was not statistically significant (chi 2 = 1.57, P = .21). The Mantel-Haenszel odds ratio, adjusting for age, however, indicated a 4.71-fold increased risk of diabetes among Mexican Americans with the CC genotype compared with Mexican Americans without this genotype (chi 2 = 5.38, P = .020). The age of onset of diabetes was also slightly younger in CC homozygote cases (45.4 +/- 9.2 yr) than in CX or XX cases (47.7 +/- 9.0 and 48.6 +/- 9.6 yr, respectively), although this difference was not statistically significant (P .467).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2568959 TI - Paradoxical effects of endogenous and exogenous insulin on amino acid transport activity in the isolated rat pancreas: somatostatin-14 inhibits insulin action. AB - Regulatory effects of insulin, somatostatin and cholecystokinin on amino acid transport in the isolated perfused rat pancreas have been studied using a rapid dual isotope dilution technique. Uni-directional L-serine transport (15 s) was quantified relative to an extracellular tracer D-mannitol over a wide range of substrate concentrations. In pancreata perfused with 2.5 mmol/l D-glucose, a weighted nonlinear regression analysis of overall transport indicated an apparent Km = 14.4 +/- 1.6 mmol/l and Vmax = 25.9 +/- 1.4 mumol.min-1.g-1 (n = 6). Although L-serine transport was stimulated during perfusion with 100 microU/ml bovine insulin, endogenous insulin (7-25 ng.min-1.g-1) released during continuous perfusion with either 8.8 mmol/l or 16.8 mmol/l D-glucose had no such effect. Exogenous somatostatin-14 (250 pg/ml) or cholecystokinin octapeptide (CCK-8, 3 x 10(-11) mol/l) appeared to increase only the Km for transport. Only CCK-8 evoked a notable protein output (2.9 +/- 0.3 mg.30 min-1.g-1) and juice flow (68 +/- 10 microliters.30 min-1.g-1, n = 3) from the exocrine pancreas. When pancreata were perfused with bovine insulin (100 microU/ml) and somatostatin-14 (250 pg/ml), the stimulatory action of exogenous insulin on L-serine transport was abolished. If endogenous insulin and somatostatin, released concurrently in response to 16.8 mmol/l D-glucose, were conveyed to the exocrine epithelium via an islet-acinar portal axis, it is conceivable that somatostatin modulates the stimulatory action of insulin on basolateral amino acid transport in the exocrine pancreas. PMID- 2568960 TI - The anterograde and retrograde infusion of glucagon antibodies suggests that A cells are vascularly perfused before D cells within the rat islet. AB - We have suggested that the order of cellular vascular perfusion within the islet is important in the regulation of islet hormone secretion. Anatomically, the A and D cells appear to be randomly dispersed throughout the mantle. Although islet capillary blood flow is known to be from the B-cell core to the A- and D-cell mantle, it has not yet been established whether the cells of the mantle may influence one another vascularly. Rat pancreata were perfused in vitro anterogradely and retrogradely with or without glucagon antibody in order to determine the order of cellular perfusion and interaction between the A and D cells in the islet mantle. Anterograde infusion of glucagon antibody did not affect insulin secretion, but rapidly decreased somatostatin secretion -46 +/- 8%, (p less than 0.005). Retrograde infusion of glucagon antibody decreased insulin secretion (-27 +/- 8%, p less than 0.005) but had no effect upon somatostatin secretion. This study not only confirms a core to mantle islet perfusion but also establishes that the A cell precedes the D cell in the terms of vascular perfusion. Thus within the islet, vascular borne insulin regulates the release of glucagon, which in turn, regulates the release of somatostatin. Somatostatin is vascularly neutral owing to its downstream position in the sequence (B to A to D) of cellular perfusion. PMID- 2568962 TI - Sources of intramitochondrial malate. AB - Liver mitochondria from rats treated with gluconeogenic hormones or subjected to vigorous exercise consume oxygen more rapidly than do mitochondria from control rats. These treatments result in elevated mitochondrial malate concentrations, which facilitate the entry of added substrate into the mitochondria. In this paper we describe experiments conducted to determine the source of the extra malate. Injections of glutamate plus alanine, two amino acids that are increased in blood after exercise and hormone treatment, caused liver mitochondrial malate to be increased. Injections of glucagon, cortisol, or both hormones elevated liver mitochondrial malate concentrations in both adrenalectomized and sham operated rats. PMID- 2568961 TI - Sensitivity of rat pancreatic A and B cells to somatostatin. AB - Islet A and B cells were purified from the rat pancreas and examined for their respective sensitivity to somatostatin. Both somatostatin-14 (S14) and -28 (S28) inhibited glucagon and insulin release through direct interactions with the corresponding cell types. A dose-dependent suppression of the secretory activities was paralleled by a reduction in cellular cyclic AMP formation with similar ED50 values for both actions. The somatostatin effects on pancreatic hormone release may thus be mediated via an inhibition of adenylate cyclase activity. In pancreatic A cells, S14 and S28 were equally potent inhibitors with ED50 values ranging from 2 x 10(-12) to 2 x 10(-11) mol/l. Pancreatic B cells exhibited a similar sensitivity to S28 as the A cells (ED50 of 2 to 5 x 10(-11) mol/l), but not to S14 (ED50 of 2 x 10(-9) mol/l). Extrapolation of these in vitro sensitivities of islet A and B cells to the in vivo situation suggests that both cell types can respond to circulating S28 levels and that A cells are sensitive to both locally and distally released S14. Islet B cells appear insensitive to the normal peripheral S14 levels but could respond to locally released somatostatin. The marked difference in the sensitivities of islet A and B cells to S14 suggest that these cell types are equipped with different somatostatin receptors. This notion was further supported by the cell-selective actions of the synthetic S14 analogues [D-Trp8, D-Cys14]S14 and desAsn5[D-Trp8, D Ser13]S14. PMID- 2568964 TI - Effect of somatostatin on plasma amino acid uptake by human pancreas. AB - We studied the effect of somatostatin on amino acid uptake by pancreatic acinar cells in 12 healthy male volunteers, aged 20-48 yr. Pancreatic amino acid uptake was assessed by measuring free plasma amino acid concentration before and during pancreatic stimulation with secretin (1 CU/kg body wt.h) and cerulein (50 ng/kg body wt.h). Pancreatic stimulation with these peptides caused a significant decrease in plasma amino acid concentrations. Somatostatin, given at the dosages of 0.15 and 1.35 micrograms/kg body wt.h, significantly diminished this decrease. The effect of the higher dose of somatostatin was more marked than that produced by the lower dose, compatible with dose dependence. The results suggest that somatostatin is able to inhibit the amino acid uptake by the pancreatic acinar cells. This inhibitory effect could be an important mechanism by which the peptide decreases pancreatic enzyme secretion. PMID- 2568963 TI - Inhibition of acid secretion by bombesin is partly mediated by release of fundic somatostatin. AB - The effect and mode of action of bombesin on gastric acid secretion was examined in the isolated, luminally perfused mouse stomach. Bombesin caused a concentration-dependent inhibition of basal and histamine-stimulated acid secretion and an increase in somatostatin secretion. Pertussis toxin was used to determine the contribution of somatostatin to the observed inhibition of acid secretion. Previous studies have shown that at a concentration of 125 ng/ml, pertussis toxin reverses completely the inhibition of acid secretion induced by exogenous and endogenous somatostatin. Preincubation of the stomach for 60 min with pertussis toxin (125 and 1250 ng/ml) reversed partially the inhibitory effect of bombesin on basal and histamine-stimulated acid secretion by 51%-59%, implying that bombesin-induced inhibition was partly mediated by release of somatostatin. The residual inhibition may represent a direct effect of bombesin on parietal cells. PMID- 2568965 TI - The inability of the rat aorta to relax to acetylcholine does not imply lack of a functional endothelium only. AB - 1. The inability of blood vessels to relax to acetylcholine is often used as the sole test of endothelial removal. 2. Following blotting of the intimal surface of the rat aorta with filter paper, the tissue did not relax to acetylcholine and the contractions to alpha 2-adrenoceptor agonists and to the lower concentrations of noradrenaline, adrenaline, phenylephrine and KCl were enhanced. 3. However the responses to the higher concentrations of noradrenaline, adrenaline, phenylephrine and KCl were reduced. 4. These results show that blotting with filter paper destroys the endothelium and some smooth muscle. PMID- 2568966 TI - Dose difference in beta-adrenergic blockers with intrinsic sympathomimetic action to block beta-adrenoceptors and induce sympathomimetic action. AB - 1. Isoprenaline, a beta-adrenergic full agonist, and carteolol, befunolol and BFE 55, beta-adrenergic blockers with an intrinsic sympathomimetic action, induced dose-related increases in heart rate when administered intravenously (i.v.). 2. The beta-partial agonists administered i.v. shifted a dose-heart rate increased response curve of isoprenaline, suggesting a competitive antagonism. 3. In carteolol, doses to block beta-adrenoceptors were found to be 400-1000 times lower than that to induce agonistic action. In BFE-55, doses to block beta adrenoceptors were equal to that to induce sympathomimetic action. The difference between doses of the befunolol to produce both actions was intermediate. 4. These results suggest that there may be a difference between doses blocking beta adrenoceptors and inducing sympathomimetic action in some beta-adrenergic partial agonists and, therefore, that there may exist beta-partial agonists which do not induce sympathomimetic action in doses blocking beta-adrenoceptors. PMID- 2568967 TI - A-23187 evoked transmitter release from rabbit pulmonary artery and its inhibition by reactivation of sodium-pump. AB - 1. The spontaneous [3H]-release has been measured from the isolated main pulmonary artery of the rabbit preloaded with [3H]noradrenaline in the presence of uptake blockers (cocaine, 3 x 10(-5) M; corticosterone, 5 x 10(-5) M). 2. The Ca-ionophore A-23187 (3 x 10(-7)-3 x 10(-5) M) increased the outflow of [3H] by a concentration dependent manner. 3. Inhibition of Na+-pump by removal of K+ from the external medium also increased the release of labelled noradrenaline. 4. In the absence of external K+, the applied A-23187 (3 x 10(-6) M; EC50) further increased the release of [3H]. 5. Reactivation of Na+-pump by readmission of K+ (5.9 mM) to the external medium abolished the [3H]-release which had previously been increased in "K+-free" solution. 6. The reactivated Na+-pump significantly inhibited the transmitter releasing action of A-23187. 7. This latter was antagonized by an increase of external Ca2+ (7.5 mM). 8. It is concluded that the reactivated Na+-pump caused re-establishment of Na+-gradient is capable to counteract the Ca-ionophore facilitated Ca2+-influx and release from internal stores, which can be antagonized by excess Ca2+. PMID- 2568968 TI - Xylazine-evoked depression of rat cerebral cortical neurons: a pharmacological study. AB - 1. Iontophoretic application of the veterinary tranquilizer xylazine suppressed the spontaneous firing of 134/137 neurons tested, an effect not antagonized by adenosine receptor and alpha-adrenergic receptor blocking agents. 2. Depressions in neuronal activity evoked by norepinephrine, gamma-aminobutyric acid or adenosine failed to be potentiated by xylazine. 3. However, the excitatory effects of acetylcholine and glutamate were both inhibited by xylazine. 4. Thus, the depressant effect of locally applied xylazine may involve a decrease in the excitability of the neuronal membrane. PMID- 2568969 TI - Pharmacological characterization of mare uterus motility with special reference to calcium antagonists and beta-2-adrenergic stimulants. AB - 1. Uterine motility was studied in vitro in the myometrial tissue obtained from pregnant and non-pregnant mares. 2. The spontaneous contractions of the preparations were not modified by tetrodotoxin, by anticholinergics, antiadrenergics, histamine H1 and H2 blockers, antiserotoninergic and opioid antagonists; but disappeared in Ca2+ and Na+ free medium. 3. beta 2-adrenergic stimulants like salbutamol and hexoprenaline and the calcium channel blockers nifedipine and verapamil were effective inhibitors of the amplitude of phasic contractions (ID50S for salbutamol and nifedipine were 7.7 nM and 14.6 nM, respectively in oestrus preparations). 4. The above data indicated that the mare myometrium contractility in vitro is very sensitive to the action of beta 2 mimetic compounds and calcium antagonists; nifedipine, in particular, seems to be a very promising alternative to beta 2 stimulants in the tocolytic therapy. PMID- 2568970 TI - Antiemetic treatment of chemotherapy-induced nausea in ovarian carcinoma patients. AB - In a prospective, randomized, double-blind trial the combination betamethasone dixyrazine was compared with high-dose metoclopramide as antiemetic treatment during combination chemotherapy (melphalan-doxorubicin and cisplatin) of ovarian carcinoma. Of 40 evaluable patients, 15 (38%) had previous experience with chemotherapy. Efficacy and side effects were recorded on patient and nurse questionnaires using the visual analog scale. Nausea and vomiting were prevented in 55% of the patients treated with melphalan-doxorubicin (Day 1) and in 36% of the patients treated with cisplatin (Day 2). Betamethasone-dixyrazine was superior to high-dose metoclopramide and prevented nausea in 76% compared to 32% during melphalan-doxorubicin therapy. Akathisia was noted in 21% and acute dystonic reactions in 2.6% during metoclopramide treatment but not in any case during betamethasone-dixyrazine therapy. PMID- 2568971 TI - [Inhibition of 24-hour acidity by nizatidine]. AB - In a randomized, double-blind, placebo-controlled, comparative cross-over study, we studied the effect of four H2-receptor antagonists on intragastric 24-hour acidity, nocturnal volume and acid output. Ten healthy male volunteers were administered 300 mg or 150 mg nizatidine, 800 mg cimetidine, 300 mg ranitidine, 40 mg famotidine, or placebo on several days, in each case at 9:000 PM. Nocturnal intragastric H+ concentration (mmol/l) (11:00 PM to 7:00 AM) was significantly reduced by all H2 blockers compared with placebo. We obtained the following inhibition rates: Cimetidine 67%; ranitidine 95%; famotidine 89%; nizatidine 80% (300 mg) and 69% (150 mg). Nocturnal acid (mmol/l) and volume output (ml/h) were also significantly (compared with placebo) inhibited by all four H2-receptor antagonists. Inhibition of nocturnal acid secretion was almost identical on nizatidine 300 mg nocte, ranitidine 300 mg nocte, famotidine 40 mg nocte, and cimetidine 800 mg nocte. Nizatidine 300 mg nocte and 150 mg nocte exclusively reduced acid secretion at night, without an aftereffect into the following day (8:00 AM to 6:00 PM). These results suggest that the clinical efficacy of these H2-receptor antagonists is identical with respect to healing peptic ulcer disease and providing freedom from pain. It is generally accepted today that gastric acid inhibitors used in the treatment of peptic ulcer disease should interfere with daytime gastric acid secretion as little as possible, particularly since the acid protects the stomach from bacteria ingested with the food during the day. PMID- 2568972 TI - [Systemic pharmacotherapy in backache. Indications and practical applications]. AB - The treatment of acute and chronic low back pain is a problem frequently encountered in the doctor's office. Selective drug therapy has a useful role to play within the framework of the overall therapeutic strategy. Indications and practical application of the major analgesic agents in various low back pain conditions are discussed. PMID- 2568973 TI - [Airway contraction and anti-asthmatic agents]. PMID- 2568976 TI - Hospitals challenged to become health center. PMID- 2568975 TI - Influence of lactate on isoproterenol-induced lipolysis and beta-adrenoceptors distribution in human fat cells. AB - The influence of lactate on human adipocytes lipolysis and the possible relationship between lactate-induced metabolic effects and beta-adrenoceptor binding sites were investigated. beta-sites were identified in membranes with (125I)-cyanopindolol and in intact cells with (125I)-cyanopindolol and (3H)-CGP 12177. Lactate reduced isoproterenol-induced lipolysis in a dose-response fashion and such inhibition became significant only at 16 mmol/l lactate. Exposure of human fat cells to 16 mmol/l lactate significantly reduced beta-adrenoceptors density on crude membranes. When the binding assay was performed on intact cells using (125I)-cyanopindolol at 37 degrees C, the radioligand identified the same number of receptors, regardless of the presence of lactate in the preincubation medium. When (3H)-CGP 12177 was used, it bound to about 35% less receptors in lactate pre-treated cells than in control. Seemingly, at 37 degrees C, because of its lipophilicity, (125I)-cyanopindolol can cross the plasma membrane and bind to intracellular sites whereas, (3H)-CGP 1277, due to its hydrophilicity, identifies surface receptors only. Thus, the present in vitro study provides evidence that high levels of lactate, similar to the concentrations usually achieved in overt lactic acidosis, are able per se to inhibit human lipolysis and to redistribute beta-adrenoceptors from cell surface to a domain not accessible to hydrophilic ligands. PMID- 2568974 TI - Central adrenoceptors and basal prolactin release in the rat. AB - The influence of the central adrenergic system on basal prolactin secretion was investigated in the rat. Several selective adrenoceptor blockers were centrally administered and their effects on prolactin secretion were observed. Blockade of beta-1 receptors by practolol, beta-2 receptors by IPS 339 and alpha-2 receptors by DG-5128 did not modify basal prolactin secretion, but alpha-1 adrenoceptor blockade by prazosin strongly enhanced prolactin plasma levels. These findings suggest that noradrenergic pathways in the central nervous system exert inhibitory tone on basal prolactin secretion, and that this effect seems to be mediated by alpha-1 adrenoceptors. PMID- 2568978 TI - Congenital systemic mastocytosis. PMID- 2568977 TI - IL-2 and IL-4 can co-modulate the generation of cytotoxic T cells through CD8- CD4- splenic lymphocytes. AB - Following activation with concanavalin A (Con A), murine T cells are able to suppress the generation of allospecific cytotoxic T lymphocytes (CTL). We have analysed the phenotype, tissue distribution, and mode of action of these cells in an effort to understand further the regulation of CTL-mediated immunity. The precursors of such cells are rare (1 cell per 70,000 spleen cells being able to suppress the generation of a particular allospecific response), but are much more abundant in the spleen than in the thymus. By the use of cytotoxic antibodies, we have been able to demonstrate that the splenic precursors of such cells are Thy 1.2+, CD4-, CD8- but, following activation with Con A, these cells acquire the CD8 marker. Cellular suppression by these lymphocytes is dramatically increased in the presence of the Th2-derived lymphokine, IL-4, whereas IL-2, the Th1 derived lymphokine, significantly augments the generation of CTL in a mixed lymphocyte culture even though relative suppression is still evident in the presence of Con A-activated lymphocytes. Suppression is not due to overcrowding of a cell culture since adding Con A-activated cells to an A anti-B + C culture often resulted in the suppression of the A anti-B response but not the A anti-C response, or vice versa. Suppression appears to require cellular interaction since supernatants from Con A-activated lymphocytes are unable to mediate suppression. Such cells may play an important intermediate role in homeostasis. PMID- 2568979 TI - Systemic adverse reactions to glaucoma medications. PMID- 2568980 TI - [Ectopic kidney tissue in the inguinal canal. Case report and review of the literature]. AB - We report a unique case of isolated ectopic tissue associated with an undescended testis. The pathological findings are outlined, and the embryology of renal and testicular development is reviewed. PMID- 2568981 TI - Epidural abscess and subdural empyema. AB - Epidural abscess and subdural empyema are serious intracranial infections that result in significant morbidity and mortality. Frequently, they are secondary to sinusitis or middle ear disease, and the bacteria involved are inhabitants of the upper respiratory tract. Symptoms may be mild and mimic the symptoms of the underlying infection. However, especially with subdural empyema, alteration in the level of consciousness and focal neurologic deficits are common. Morbidity and mortality are minimized by early diagnosis, which is best made with computed tomography scanning, and proper therapy, which consists of surgical drainage and administration of appropriate antimicrobials. It is important that primary care physicians be aware of the clinical features of these potentially fatal complications of common infections. PMID- 2568982 TI - L-asparaginase for treatment of lymphoid neoplasia in dogs. PMID- 2568983 TI - Immunocytochemical identification and localization of the Mr 22,000 calcium binding protein (sorcin) in an adriamycin-resistant myelogenous leukemia cell line. AB - Monoclonal antibody against the Mr 22,000 calcium-binding protein (sorcin) from an adriamycin-resistant myelogenous leukemia cell line K562 (K562/ADM) was prepared and used as a probe to study the localization of sorcin in K562/ADM cells and the parental cell line, K562. Analysis of extracts from K562/ADM cells by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorescence image analysis showed that K562/ADM cells possessed abundant sorcin in the cytoplasm which was almost entirely absent from the drug-sensitive parental cell line, K562. Furthermore, immuno-electron microscopic studies revealed that sorcin was closely associated with free ribosomes, rough endoplasmic reticulum, mitochondria, microfilament bundles and perinuclear membranes. These observations provide the first clue that the Ca-binding protein, sorcin, may play an important role in the development of the multidrug resistance phenomenon, although the relationship between sorcin and P-glycoprotein is still unknown. PMID- 2568984 TI - Potentiation of some anticancer agents by dipyridamole against drug-sensitive and drug-resistant cancer cell lines. AB - In this study, we have used two different vincristine (VCR)-resistant variants, VJ-300 and HC-7-5/VCR. VJ-300 was isolated from a human cancer KB cell line and HC-7-5/VCR from a human cancer HC-7-5 cell line. VJ-300 and HC-7-5/VCR are both multidrug-resistant (MDR) variants, showing resistance to multiple anticancer drugs such as VCR, adriamycin, actinomycin D and daunomycin. Dipyridamole, a specific inhibitor of nucleoside transport, potentiated these anticancer drugs about 2- to 10-fold against KB and VJ-300. Dipyridamole almost completely reversed drug resistance to actinomycin D in VJ-300 cells with about a 70-fold higher resistance to actinomycin D. Dipyridamole inhibited the efflux of actinomycin D and VCR from VJ-300 cells. Dipyridamole enhanced the uptake of VCR but not that of actinomycin D in VJ-300 and KB. Dipyridamole at 10-100 microM inhibited photoaffinity labeling with [3H]azidopine of the cell-surface protein P glycoprotein in VJ-300 cells. Dipyridamole potentiated 5-fluorouracil and hexylcarbamoyl-5-fluorouracil in cultured KB and VJ-300, but it annihilated the cytotoxic action of 5-fluorouridine. Potentiation of 5-fluorouracil by dipyridamole against HC-7-5 and HC-7-5/VCR was also observed, but appeared to be less than in VJ-300 and KB cells. Dipyridamole almost completely inhibited the cellular accumulation of 5-fluorouridine, but not that of 5-fluorouracil. Thus, dipyridamole appeared to potentiate anticancer agents through pleiotropic action sites, one of which is inhibition of enhanced efflux of MDR cell lines and the other of which is inhibition of nucleoside transport. Dipyridamole might be a useful and potent agent to potentiate anticancer agents and reverse drug resistance. PMID- 2568985 TI - Molecular characterization of a fimbrial adhesin, F1845, mediating diffuse adherence of diarrhea-associated Escherichia coli to HEp-2 cells. AB - A fimbrial adhesin, designated F1845, was found to be responsible for the diffuse HEp-2 cell adherence of a diarrheal Escherichia coli isolate. The genetic determinant of F1845 was cloned, and the order of the genes necessary for production of F1845 was determined by maxicell analysis. Five polypeptides with apparent sizes of 10, 95, 27, 15.5, and 14.3 kilodaltons (kDa) were found to be encoded in that order by the F1845 determinant. The nucleotide sequence of the 14.3-kDa subunit gene was determined and found to share extensive homology in its signal sequence with the gene encoding the structural subunit of the AFA-1 hemagglutinin of a uropathogenic E. coli strain (A. Labigne-Roussel, M.A. Schmidt, W. Walz, and S. Falkow, J. Bacteriol. 162:1285-1292, 1985) but not in the region encoding the mature protein. Southern blot hybridizations indicated that the F1845 determinants are of chromosomal origin. Hybridization studies using a probe from the region encoding the 95-kDa polypeptide indicated that related sequences may be plasmid associated in some strains and chromosomal in others. Additional hybridization studies of E. coli isolates possessing sequence homology to the F1845 determinant suggest that the sequences in the 5' region of the F1845 structural subunit gene are more highly conserved than sequences in the 3' region. PMID- 2568987 TI - Differential stimulation of rat lung particulate guanylate cyclase activity by atrial natriuretic peptide and sodium nitroprusside. AB - The effects of alpha-rat atrial natriuretic peptide (alpha-rANP) and sodium nitroprusside on the activity of rat lung particulate guanylate cyclase were examined. The particulate guanylate cyclase in partially purified rat lung membranes was stimulated by both alpha-rANP and nitroprusside. The effects of alpha-rANP and nitroprusside were, however, not additive. Diamide and N ethylmaleimide almost completely abolished the nitroprusside-mediated stimulation, while they had only moderate effects on the alpha-rANP-mediated stimulation of the enzyme activity. ATP potentiated the enzyme stimulation by alpha-rANP, whereas it had no effect on the nitroprusside-mediated stimulation. These findings suggest that the stimulation of lung particulate guanylate cyclase activity by alpha-rANP and nitroprusside is mediated by different mechanisms. PMID- 2568986 TI - Lysis of Escherichia coli by the bacteriophage phi X174 E protein: inhibition of lysis by heat shock proteins. AB - Lysis of Escherichia coli by the cloned E protein of bacteriophage phi X174 was more rapid than expected when bacteria were shifted from 30 to 42 degrees C at the time of E induction. Since such treatment also induces the heat shock response, we investigated the effect of heat shock proteins on lysis. An rpoH mutant was more sensitive to lysis by E, but a secondary suppressor mutation restored lysis resistance to parental levels, which suggests that the sigma 32 subunit itself did not directly increase lysis resistance. At 30 degrees C, mutants in five heat shock genes (dnaK, dnaJ, groEL, groES, and grpE) were more sensitive to lysis than were their wild-type parents. The magnitude of lysis sensitivity varied with mutation and strain background, with dnaK, dnaJ, and groES mutants consistently exhibiting the greatest sensitivities. Extended protection against lysis occurred when overproduction of heat shock proteins was induced artificially in cells that contained a plasmid with the rpoH+ gene under control of the tac promoter. This protective effect was completely abolished by mutations in dnaK, dnaJ, or groES but not by grpE or groEL mutations. Altered membrane behavior probably explains the contradiction whereby an actual temperature shift sensitized cells to lysis, but production of heat shock proteins exhibited protective effects. The results demonstrate that E-induced lysis can be divided into two distinct operations which may now be studied separately. They also emphasize a role for heat shock proteins under non-heat shock conditions and suggest cautious interpretation of lysis phenomena in systems where E protein production is under control of a temperature-sensitive repressor. PMID- 2568988 TI - Comparative study of the sugar chains of gamma-glutamyltranspeptidases purified from human hepatocellular carcinoma and from human liver. AB - The sugar chains of gamma-glutamyltranspeptidases, purified from human hepatoma and from normal human liver, were released quantitatively as oligosaccharides by hydrazinolysis. Comparative study of their structures revealed that the following structural alterations are induced by hepatocyte carcinogenesis: 1) high mannose type sugar chains are detected in the hepatoma enzyme but not in the normal liver enzyme; 2) abnormal biantennary sugar chains containing C-2,4 outer chain branches newly appeared; 3) the total amounts of tri- and tetraantennary sugar chains containing C-2,6 outer chain branches increased up to three times. PMID- 2568989 TI - Binding of D-phenylalanine and D-tyrosine to carboxypeptidase A. AB - The structures of the complexes of carboxypeptidase A with the amino acids D phenylalanine and D-tyrosine are reported as determined by x-ray crystallographic methods to a resolution of 2.0 A. In each individual study one molecule of amino acids binds to the enzyme in the COOH-terminal hydrophobic pocket: the carboxylate of the bound ligand salt links with Arg-145, and the alpha-amino group salt links with Glu-270. The carboxylate of Glu-270 must break its hydrogen bond with the native zinc-bound water molecule in order to exploit the latter interaction. This result is in accord with spectroscopic studies which indicate that the binding of D or L amino acids (or analogues thereof) allows for more facile displacement of the metal-bound water by anions (Bicknell, R., Schaffer, A., Bertini, I., Luchinat, C., Vallee, B. L., and Auld, D. S. (1988) Biochemistry 27, 1050-1057). Additionally, we observe a significant movement of the zinc-bound water molecule (approximately 1 A) upon the binding of D-ligands. We propose that this unanticipated movement also contributes to anion sensitivity. The structural results of the current x-ray study correct predictions made in an early model building study regarding the binding of D-phenylalanine (Lipscomb, W. N., Hartsuck, J. A., Reeke, G. N., Jr., Quiocho, F. A., Bethge, P. H., Ludwig, M. L., Steitz, T. A., Muirhead, H., and Coppola, J. C. (1968) Brookhaven Symp. Biol. 21, 24-90). PMID- 2568990 TI - Vicinal diamines as pyrroloquinoline quinone-directed irreversible inhibitors of lysyl oxidase. AB - The observation that aliphatic diamines become poor substrates as the carbon chain length decreases and that ethylenediamine, the shortest diamine, is an irreversible inhibitor of lysyl oxidase led to the investigation of the mechanism of inhibition by ethylenediamine. The cis but not the trans isomer of 1,2 diaminocyclohexane was also a potent irreversible inhibitor of lysyl oxidase, consistent with the interaction of both amino groups of vicinal diamines with an enzyme moiety. Both cis-1,2-diaminocyclohexane and ethylenediamine but not trans 1,2-diaminocyclohexane markedly perturbed the spectrum of free pyrroloquinoline quinone (PQQ), a covalently linked form of which is the carbonyl cofactor of lysyl oxidase. cis-1,2-Diaminocyclohexane also induced similar changes in the spectrum of lysyl oxidase. The perturbations of the spectra of PQQ or of lysyl oxidase by cis-1,2-diaminocyclohexane or ethylenediamine as well as the development of irreversible inhibition of the enzyme by cis-1,2 diaminocyclohexane or ethylenediamine were all markedly reduced under anaerobic conditions. Moreover, approximately 1 mol of H2O2 was released per mol of PQQ or lysyl oxidase upon aerobic incubation with cis-1,2-diaminocyclohexane, while approximately 2 mol of 3H+ were released from cis-[1,2-3H] 1,2-diaminocyclohexane per mol of PQQ or lysyl oxidase under corresponding conditions. A proposal for the mechanism of inhibition of lysyl oxidase by vicinal diamines is presented which involves limited oxidation of the diamine linked to PQQ at the active site so that the PQQ-diamine complex is finally stabilized by a conjugated 6-membered ring. PMID- 2568992 TI - Bacteriorhodopsin mutants of Halobacterium sp. GRB. II. Characterization of mutants. AB - The bacterioopsin genes of Halobacterium sp. GRB (Ebert, K., Goebel, W., and Pfeifer, F. (1984) Mol. & Gen. Genet. 194, 91-97) wild type and 10 independent mutants of different phenotypes have been cloned and sequenced. The wild type gene has two conservative changes compared to the gene of Halobacterium halobium, so that the proteins of the two species are identical. Six different mutations at five different codons have been found, leading to the following amino acid changes compared to the wild type: Trp10----Cys (three cases), Tyr57----Asn, Asp85----Glu, Asp06----Asn (three cases), Asp96----Gly, Trp138----Arg. A first characterization of the mutant proteins is given, and their implications for models of bacteriorhodopsin structure and function are discussed. PMID- 2568991 TI - Evidence of alpha 1-adrenergic----protein kinase C----Na+/H+ antiporter-dependent increase in pinealocyte intracellular pH. Role in the adrenergic stimulation of cGMP accumulation. AB - The regulation of intracellular pH (pHi) in isolated rat pinealocytes was studied using the fluorescent pH indicator 2',7'-bis(carboxyethyl)-5(6) carboxyfluorescein. Resting pHi was 7.09 when the extracellular pH (pHe) was 7.2. Treatment of pinealocytes with the physiological regulator of pineal function, norepinephrine, resulted in a concentration-dependent increase in pHi. Further analysis indicated that norepinephrine is probably acting via an alpha 1 adrenergic----[Ca2+]i----Ca2+/phospholipid- dependent protein kinase (protein kinase C) mechanism to activate the Na+/H+ antiporter, thereby causing cytoplasmic alkalization. A potential influence of cytosolic alkalization on the responsiveness of cyclic nucleotides to adrenergic agonists was also studied. Five analogs of the antiporter inhibitor amiloride reduced norepinephrine stimulation of cGMP accumulation with the same relative potency as they act on the antiporter. In contrast, although inhibitory effects of these compounds on cAMP accumulation were detectable, they occurred at 10-100-fold higher concentrations, and the relative potency of these inhibitors did not indicate they were acting via the antiporter. These findings provide evidence that 1) alpha 1-adrenergic receptor activation increases pinealocyte pHi through Ca2+--- protein kinase C-dependent activation of the Na+/H+ antiporter; and 2) norepinephrine stimulation of cGMP accumulation is pHi-dependent. It would appear that alpha 1-adrenergic regulation of pHi via the Na+/H+ antiporter may be of general importance in the control of cGMP accumulation. PMID- 2568993 TI - Size-exclusion chromatography of DNA restriction fragments. Fragment length determinations and a comparison with the behaviour of proteins in size-exclusion chromatography. AB - Size-exclusion chromatography of double-stranded DNA restriction fragments on Superose 6 is shown to be an accurate method for chain length determination of unknown DNA. Ionic interaction was observed between DNA and the gel matrix but was easily prevented by the addition of 0.15-0.2 M sodium chloride to the eluent. Compared to protein, the selectivity curve of DNA fragments was found to be steeper reflecting the different chromatographic behaviour of rod-like and globular molecules. The relationships between the selectivity curves of DNA and protein were similar on Superose 6 and on Sephacryl S-500. PMID- 2568994 TI - High-performance liquid chromatographic determination of the enantiomers of beta adrenoceptor blocking agents in biological fluids. II. Studies with atenolol. PMID- 2568995 TI - Evidence for cholinergic modulation of aldosterone secretion in man. AB - Acetylcholine stimulates aldosterone secretion in bovine glomerulosa cells in vitro via specific cholinergic receptors. In this study we examined the effect of peripheral muscarinic blockade with atropine on metoclopramide-, angiotensin-II-, and ACTH-stimulated aldosterone secretion in man. Atropine (0.6 mg, iv) administered 10 min before MCP delayed the onset of the plasma aldosterone response and attenuated the mean peak response from 502 +/- 103 (+/- SE) to 322 +/- 72 pmol/L (P less than 0.05) without affecting zero time mean plasma aldosterone levels (144 +/- 28 vs. 136 +/- 36 pmol/L for control and atropine, respectively). This inhibitory effect was not mediated by changes in PRA or plasma potassium or ACTH (as reflected by cortisol) concentrations. Atropine also attenuated the plasma aldosterone response to a low dose angiotensin II infusion (2 ng/kg.min; control, 449 +/- 99 pmol/L; atropine, 297 +/- 78 pmol/L; P less than 0.05). In contrast, atropine had no effect on the plasma aldosterone response to a bolus dose (250 micrograms) of ACTH. Neither atropine (0.6 mg, iv) nor the cholinergic muscarinic agonist bethanechol (5 mg, sc) alone elicited a change in plasma aldosterone. Collectively, these data provide evidence for cholinergic modulation of aldosterone secretion in man. We conclude that cholinergic mechanisms may facilitate the aldosterone responses to angiotensin-II and metoclopramide, but not to ACTH. PMID- 2568997 TI - Evidence from neuron/lymphoma heterokaryons for a common trans-acting factor suppressing Thy-1 expression. AB - We have investigated the tissue specificity of a trans-acting regulatory factor observed to suppress Thy-1 expression during normal neuronal differentiation. Heterokaryons were constructed between Thy-1.1+ expressing mouse T cell lymphoma cells and Thy-1.2- mouse sensory neurons and their surface phenotypes determined by immunofluorescence histochemistry 16 h after fusion. Thy-1.1 expression was observed to be specifically suppressed in such heterokaryons whilst the expression of the lymphoma cell surface marker, Ly-1, was not altered. As the nuclei did not fuse in the heterokaryons, it appears that the developmentally regulated diffusible suppressor factor active in sensory neurons is not tissue specific and can down-regulate Thy-1 expression in lymphoma cells. We also report the suppression of the H-2Kk surface antigen in heterokaryons by a similar but independent regulatory mechanism. PMID- 2568999 TI - Identification of vagal efferent fibers and putative target neurons in the enteric nervous system of the rat. AB - The stomach and small intestine receive an efferent innervation from the dorsal motor nucleus of the vagus (DMX). The current experiments were undertaken as a partial test of the hypothesis that the CNS innervates only a small number of command neurons in a restricted number of enteric ganglia. The anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L) was injected into the DMX by iontophoresis, and 10-21 days later PHA-L was visualized in the bowel by immunofluorescence. Varicose vagal efferent fibers, labeled by PHA-L, were found in the myenteric plexus as far distally as the ileo-colic junction. PHA-L-labeled varicose axons were rare in comparison to nonlabeled fibers, entered a minority of myenteric ganglia, and contacted a small proportion of the neurons. Ganglia thus innervated by vagal efferent fibers were more numerous in the stomach than in the small intestine. Within the stomach, these ganglia were common in the antrum than in the corpus and none were found in the wall of the rumen. Innervated ganglia in the small intestine became progressively more sparse distally. No PHA-L-labeled axons were observed in the submucosal plexus, thus raising the possibility that vagal modulation of secretomotor responses involves an intermediate synapse in the myenteric plexus. Nonvaricose bundles of PHA-L labeled fibers were also observed. These bundles appeared to utilize the connectives of the myenteric plexus as a pathway within which to descend within the bowel. Vagal efferent bundles were found to pass through the pyloric sphincter to enter the small intestine from the stomach; thus vagal fibers can reach the distal intestine by an intraenteric route that is not lesioned by crushing mesenteric nerves. The existence of this pathway affects the interpretation of experiments seeking to utilize such lesions to distinguish intrinsic from extrinsic neurites. Possible target neurons of the vagal efferent innervation were identified by simultaneously demonstrating the immunoreactivities of 5-hydroxytryptamine (5-HT), vasoactive intestinal polypeptide (VIP), enkephalin (ENK), galanin (GAL), and tyrosine hydroxylase (TH) along with that of PHA-L. Vagal terminals in the myenteric plexus appeared selectively to contact 5-HT- and, to a significantly lesser extent, VIP-, but not ENK- or GAL-immunoreactive neurons. Apparent vagal innervation of 5-HT immunoreactive neurons was significantly more common in the duodenum, where a majority of the 5-HT-immunoreactive cells were encircled by varicose PHA-L labeled axons, than in the stomach.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2568998 TI - Distribution of somatostatin-like immunoreactivity in the monkey amygdala. AB - The distribution of somatostatin-like immunoreactivity was studied in the macaque monkey (Macaca fascicularis) by using primary antisera that recognize somatostatin-28 (S309) or somatostatin-28(1-12) (S320). Somatostatin immunoreactive neuronal cell bodies were observed in all amygdaloid nuclei and cortical regions. The density of labeled cells varied substantially, however, both within and across the various amygdaloid subdivisions. The highest densities of labeled neurons were observed in layer III of the periamygdaloid cortex, in layers II and III of the medial nucleus, in the magnocellular division of the accessory basal nucleus, and in the medial portion of the lateral nucleus. Many labeled cells were also consistently observed in the caudoventral portion of the lateral division of the central nucleus. Labeled cells were heterogeneous in size and shape ranging from small and spherical to large and multipolar. The density of somatostatin-immunoreactive fibers also varied greatly from region to region and was often inversely related to the density of immunoreactive cells. Highest densities of immunoreactive fibers were observed in the periamygdaloid cortex, medial nucleus, parvicellular division of the accessory basal nucleus, paralaminar nucleus, ventrolateral portion of the lateral nucleus, parvicellular division of the basal nucleus, and the lateral division of the central nucleus. Fibers and terminals in the central nucleus had a coarsely varicose appearance and this pattern of staining was continuous along the trajectory of the central nucleus projection to the bed nucleus of the stria terminalis. The large, immunoreactive varicosities located in this area often appeared to outline dendritic or vascular profiles within the substantia innominata. The lowest levels of somatostatin-immunoreactive fibers were observed in the magnocellular division of the basal nucleus and in the ventromedial portion of the accessory basal nucleus. PMID- 2569000 TI - Chronic pruritic eruption in patients with acquired immunodeficiency syndrome associated with increased antibody titers to mosquito salivary gland antigens. AB - Five of seven patients with acquired immunodeficiency syndrome (AIDS) who had pruritus and a chronic, nonspecific-appearing skin eruption had increased antibody titers to antigens in the salivary glands of Aedes taeniorhynchus, a salt marsh mosquito common to South Florida. We hypothesize that the pruritus and skin lesions in patients with AIDS represent a form of chronic "recall" reaction. Increased antibody titers to mosquito salivary gland antigens may be a consequence of nonspecific B cell activation, a feature of AIDS. PMID- 2568996 TI - Enteropathogenic Escherichia coli serotype O111:HNT isolated from preterm neonates in Nairobi, Kenya. AB - This investigation was initiated as a consequence of several cases of diarrhea in a nursery ward for preterm babies in Nairobi, Kenya. Ten lactose-positive colonies were isolated from the stools of each of 30 neonates, regardless of whether they had diarrhea; 229 strains were identified as Escherichia coli and 65 strains were identified as Klebsiella pneumoniae. Six strains were lost during laboratory handling. No other bacterial, viral, or parasitic enteropathogens were identified. Using synthetic alkaline phosphatase-labeled probes, the bacterial isolates were found to be negative for the presence of genes coding for heat stable and heat-labile enterotoxins. Seventy-eight E. coli strains isolated from a total of 13 neonates possessed the E. coli enteropathogenic adhesion factor (EAF) gene, as demonstrated by the use of a cloned radiolabeled DNA fragment probe. These strains possessed similar plasmid profiles constituting a core plasmid profile, and while all adhered to HeLa cells, none produced Vero cell cytotoxins. The EAF gene was located on a 65-megadalton plasmid. Serotyping showed the strains to be of serogroup O111 and serotype H nontypable, a well known enteropathogenic type. Five neonates died during the outbreak, and the fatality rate was 30.7% (4 of 13) for neonates infected with EAF-positive E. coli strains compared with 7.7% (1 of 13) for neonates from whom only EAF-negative E. coli strains were isolated. K. pneumoniae only was isolated from five neonates. PMID- 2569001 TI - Myocardial beta-adrenoceptor changes in heart failure: concomitant reduction in beta 1- and beta 2-adrenoceptor function related to the degree of heart failure in patients with mitral valve disease. AB - In patients suffering from end-stage congestive cardiomyopathy, cardiac beta 1 adrenoceptor function is markedly reduced, whereas cardiac beta 2-adrenoceptor function is nearly normal. To determine whether beta 1-adrenoceptor function is impaired in heart failure selectively, beta 1- and beta 2-adrenoceptor density and functional responsiveness in the right and left atria and the left papillary muscles from patients with mitral valve disease (functional class III to IV) were studied. In all three tissues concomitantly beta 1- and beta 2-adrenoceptor density gradually declined when the degree of heart failure increased from functional class III to IV. This decrease in beta 1- and beta 2-adrenoceptor density was accompanied by similar decreases in the contractile response of isolated electrically driven right atrial and left ventricular papillary muscles to beta-adrenergic agonists. It is concluded that a decrease in cardiac beta adrenoceptor function is a general phenomenon in heart failure, and its extent is related to the degree of heart failure. However, in contrast to congestive cardiomyopathy, in mitral valve disease the decrease in cardiac beta-adrenoceptor function is due to a concomitant decrease in beta 1- and beta 2-adrenoceptors. PMID- 2569002 TI - Beta-adrenergic blockers, immunotherapy, and skin testing. American Academy of Allergy and Immunology. PMID- 2569003 TI - A monoclonal antibody directed against quail tyrosine hydroxylase: description and use in immunocytochemical studies on differentiating neural crest cells. AB - Catecholamine (CA) synthesis is one of the phenotypic traits expressed by some neural crest-derived cells in vivo and in vitro. In the present study, we have evidenced, in quail embryos, the expression of the first enzyme of CA metabolism, tyrosine hydroxylase (TOH), using a monoclonal antibody raised against the quail enzyme. This antibody also recognizes TOH from chick and pleurodele, but not from several mammalian species (rat, human). We have also investigated the extent to which TOH-positive cells, differentiated in neural crest cultures, express structural neuronal markers and display vasoactive intestinal polypeptide (VIP) and substance P (SP) immunoreactivity. Double-immunolabeling experiments show that, in vitro, half of the population of TOH-positive cells exhibits tetanus toxin binding sites but none of them are recognized by a neurofilament antibody. On the other hand, some TOH-positive cells contain VIP or SP. These observations suggest that under our culture conditions autonomic neural crest precursors differentiate only into immature sympathoblasts, but are able to synthesize peptides in addition to CA. PMID- 2569004 TI - The interaction between mast cells and endothelial cells. AB - Mast cells (MC) are widely distributed throughout different organs with a relative predilection for potential portals of entry into the host. In tissues, MC are generally concentrated around small blood vessels and lymphatics, as well as nerves and glandular tissue. This close association with vascular structures suggests an important interaction between MC and endothelial cells (EC). Tissue MC are known to generate and release a number of mediators including histamine, prostaglandin D2, and leukotrienes that induce vasodilatation and increased vascular permeability. These MC-mediated effects on vessels may enhance responses to tissue injury or infection by facilitating the deposition of plasma components and inflammatory cells into involved sites. Important interactions between MC, EC, and peripheral nerves also occur. MC-derived histamine and possibly other mediators induce axon reflexes in unmyelinated sensory nerves leading to the release of neurotransmitters such as substance P, calcitonin gene-related peptide, and adenine nucleotides. These neurotransmitters exert direct effects on blood vessels, and in some instances, may act as MC stimulators. In vitro studies indicate that MC also affect EC growth and new blood vessel formation. Mast cell derived heparin has been implicated as an important cofactor in tumor-induced angiogenesis, whereas histamine has been reported to augment human dermal EC growth in culture. The recent identification of a tumor necrosis factor-like peptide in MC and the reported cytostatic effects of TNF on EC suggest that MC may inhibit the proliferation of these cells under certain conditions. Taken together, these observations indicate that the interactions between MC and EC are important in both physiologic and pathologic events. PMID- 2569005 TI - Calculation of initial yields of single- and double-strand breaks in cell nuclei from electrons, protons and alpha particles. AB - A model of the DNA and electron and ion track structure computer codes are used to model damage in the DNA by direct action. This damage is converted into single strand breaks using the method described by Charlton and Humm (1988) in which a minimum energy deposited in a critical volume of the DNA is correlated with the production of single-strand breaks. It is then assumed that if these single strand breaks lie on opposite strands and are separated by less than a few base pairs they produce double-strand breaks. Absolute yields of both single- and double-strand breaks expressed in breaks/Gy-dalton are calculated and compared to measured yields. Good agreement is obtained for single-strand breaks while the calculated yields for double-strand breaks are greater than those measured. PMID- 2569006 TI - Induction of the adaptive response by X-rays is dependent on radiation intensity. AB - Human lymphocytes pretreated with low (0.01 Gy) but not high (0.5 Gy) doses of X rays become somewhat refractory to the induction of chromatid deletions by subsequent exposure to high (1.5 Gy) doses of X-rays (i.e. the yield of chromatid deletions is less than the sum of the yields induced by the pre-exposure and the subsequent challenge doses). This adaptive response can also be induced by pretreating the cells with very low, or even high, concentrations of tritiated thymidine. Because high concentrations of tritiated thymidine result in high doses of radiation that are delivered at very low dose-rates (i.e. less than 0.01 Gy/min), the lack of adaptation following high pre-treatment doses of X-rays could be attributed to their higher dose-rates. To test the effect of X-ray intensity on the induction of the adaptive response, lymphocytes were irradiated with 0.5 Gy of X-rays at 0.005-0.5 Gy/min at 28-30 h of culture, and then irradiated with 1.5 Gy at 48 h. Chromatid deletions were measured 6 h later. The results show that 0.5 Gy of X-rays given at low dose-rates (0.005 or 0.01 Gy/min), but not at high dose-rates (0.1, 0.2, or 0.5 Gy/min), are capable of inducing the adaptive response. Furthermore, experiments in which a male subject's cells exposed to 0.5 Gy given at 0.005 Gy/min were cocultivated with a female subject's cells irradiated with 0.5 Gy at 0.5 Gy/min showed that cells exposed to radiation at low and high intensity progress to metaphase equally and, therefore, that the lack of an adaptive response at high dose-rates cannot be attributed to selection of radioresistant cells. Although the induction of the adaptive response at higher X-ray doses occurs at low radiation intensity, there seems to be a minimum dose required for this effect; e.g., 0.01-Gy pretreatments induced the adaptive response when given at 0.2 Gy/min, but not at 0.005 Gy/min. Thus, the adaptive response is dependent both on the total dose of the pretreatment and on the rate at which the dose is given. PMID- 2569007 TI - 'Chemical repair' in irradiated DNA solutions containing thiols and/or disulphides. Further evidence for disulphide radical anions acting as electron donors. AB - The ring-closed disulphide radical anion D/SS.-, formed radiolytically either by hydrogen abstraction from dithiothreitol (D(SH)2), or by one-electron reduction of the corresponding cyclic dithiane (DS2), is proposed to engage efficiently in 'chemical repair' of .OH-induced DNA intermediates (DNA.) under gamma-irradiation of aqueous DNA solutions, DNA.+D/SS.- ----DNA+DS2 Evidence for this concept derives from the observation that radioprotection of DNA by DS2 or D(SH)2 can be enhanced in anoxic N2O-saturated solution by the addition of formate, at a constant total .OH scavenger capacity: carbon dioxide radical anions (CO2.-) actually promote the generation of D/SS.-, by interacting both with DS2 and D(SH)2. Oxygen enhancement of radiation-induced DNA damage in systems containing DS2 and/or D(SH)2 can be easily explained with the above concept, as previously proposed in the case of cysteine (Prutz and Monig 1987), by O2-induced annihilation of the protective disulphide radical anion. In further support of the mechanism proposed it is shown that the protection efficiency under anoxia, and concomitantly the oxygen enhancement, increase with dose rate and with salt concentration, when DNA is irradiated in presence of DS2. PMID- 2569008 TI - Rapid human chromosome aberration analysis using fluorescence in situ hybridization. AB - We have used in situ hybridization of repeat-sequence DNA probes, specific to the paracentromric locus 1q12 and the telomeric locus 1p36, to fluorescently stain regions that flank human chromosome 1p. This procedure was used for fast detection of structural aberrations involving human chromosome 1p in two separate experiments. In one, human lymphocytes were irradiated with 0, 0.8, 1.6, 2.4 and 3.2 Gy of 137Cs gamma-rays. In the other, human lymphocytes were irradiated with 0, 0.09, 0.18, 2.0, 3.1 and 4.1 Gy of 60Co gamma-rays. The frequencies (per cell) of translocations and dicentrics with one breakpoint in 1p and one elsewhere in the genome were determined for cells irradiated at each dose point. These frequencies both increased with dose, D, in a linear-quadratic manner. The delta, alpha, and beta coefficients resulting from a fit of the equation f(D)=delta + alphaD + betaD2 to the translocation frequency dose-response data were 0.0025, 0.0027 and 0.0037 for 137Cs gamma-rays, and 0.0010, 0.0041, and 0.0057 for 60Co gamma-rays. The delta, alpha, and beta coefficients resulting from a fit to the dicentric frequency dose-response data were 0.0005, 0.0010 and 0.0028 for 137Cs gamma-rays and 0.0001, 0.0002 and 0.0035, for 60Co gamma-rays. Approximately 32,000 metaphase spreads were scored in this study. The average analysis rate was over two metaphase spreads per minute. However, an experienced analyst was able to find and score one metaphase spread every 10s. The importance of this new cytogenetic analysis technique for biological dosimetry and in vivo risk assessment is discussed. PMID- 2569009 TI - Does thermotolerance occur in human red cells? AB - Osmotic fragility and autohemolysis were used as endpoints in the measurement of damage to the plasma membrane in human erythrocytes, after single or double heat treatments. Injury recorded above 46 degrees C, and the induction of thermotolerance in the enucleate cells after a primary heat treatment of 44 degrees C for 15 min, indicates that the plasma membrane plays an important role in heat injury and in thermotolerance. PMID- 2569010 TI - Polyamines in the small intestine of rats after whole-body irradiation. AB - The behaviour of polyamines (putrescine, spermidine and spermine) in the small intestine was studied in rats exposed to a single whole-body dose of 3 Gy administered at four different times of the day. Polyamine content decreased a few hours after irradiation but quickly returned to normal values. Putrescine levels reached the lowest values and returned to the control levels later than spermidine and spermine. A temporary increase in spermidine and spermine content was observed from 5 to 20 days. The postirradiation behaviour of the animals exposed at different times of the day showed no important differences. Polyamine modifications were studied in relation to [3H]thymidine uptake and other morphological and biochemical parameters. The results indicate that polyamine content can be used to monitor the damage and recovery phases of radiation injury in the small intestine. PMID- 2569011 TI - Treatment of wound sepsis in irradiated mice. AB - Infections in the immunocompromised host are difficult to treat. The local and systemic effect of penicillin therapy, supplemented by immunoglobulins, and pentoxifylline on wounds infected by Staphylococcus aureus was evaluated in mice irradiated with 6.5 Gy 60Co gamma-rays. Three days after irradiation a suspension of S. aureus was inoculated subcutaneously over the gluteus muscle of anesthetized mice. The skin and the muscle were incised at the site of the inoculation. Treatment with 62.5 mg/kg penicillin-G was administered for 10 days. Numbers of bacteria per mg muscle and presence of organisms in spleens and livers were determined. Numbers of bacteria were significantly reduced from 7.3 (+/- 0.3) to 5.3 (+/- 0.4) log10 CFU/mg (+/- SEM) muscle in treated animals. Administration of immunoglobulin G i.v. or pentoxifylline i.p. alone, or in addition to penicillin-G, did not further reduce the number of bacteria. Increase in the dose of penicillin to 250 mg/kg decreased the number of bacteria more than 62.5 mg/kg. Bacteria were recovered from spleens and/or livers of all 13 untreated mice, and only in six of the 13 penicillin-treated mice (P less than 0.05). Penicillin therapy reduced the systemic spread of S. aureus. The model provides a means to evaluate regimens for treatment of bacterial wound infections in irradiated animals. The data illustrated the ability of antimicrobial agents to contain but not cure the infection in the immunocompromised host, and the lack of efficacy of immunoglobulins in neutropenic mice. PMID- 2569012 TI - Radiation nephropathy in mature pigs following the irradiation of both kidneys. AB - Both kidneys in mature female Large White pigs, ca. 45 weeks old were irradiated with single doses of 7.8-14.0 Gy of 60Co gamma-rays. Radiation-induced changes in renal function were assessed on the basis of sequential individual measurements of kidney glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) for up to 24 weeks after irradiation. At the same time intervals blood samples were taken to determine plasma renin levels and the haematological status of each animal. Two weeks after irradiation the ERPF and in particular the GFR was increased. This was followed by a pronounced, dose-dependent, decline in renal haemodynamics. Minimal functional levels were observed 8-12 weeks after irradiation. Function then somewhat recovered. There was a significant (r greater than or equal to 0.98; p less than 0.001) inverse relationship between the mean values of GFR and ERPF, determined at 4-24 weeks after irradiation, and the radiation dose. After a dose of 7.8 Gy the reduction in ERPF was greater than that for GFR. However, at higher doses both parameters were reduced to an equal extent. The resulting slope of the dose-effect curve for impaired renal function versus dose was significantly steeper for GFR than that for ERPF (p less than 0.001). There was a significant reduction (p less than 0.002) in the erythrocyte count, the haematocrit and haemoglobin levels within 6-8 weeks of irradiation; this anaemia was characterized as a normochromic normocytic anaemia. There were no marked changes in plasma renin levels as a result of renal irradiation. The pathogenesis of late radiation-induced damage to the kidney is discussed in the light of these findings. PMID- 2569013 TI - The effect of age and the proportion of renal tissue irradiated on the apparent radiosensitivity of the pig kidney. AB - In 14-week-old (immature) and 45-week-old (mature) pigs either the right kidney (unilateral irradiation, UI) or both kidneys (bilateral irradiation, BI) were irradiated. The kidneys of immature pigs received single doses of 7.0-12.6 Gy of 250 kV X-rays; mature pigs received single doses of 7.8-14.0 Gy of 60Co gamma rays. These were assumed to be approximately equivalent to the X-ray doses using a RBE of 0.9 gamma-rays/X-rays. The glomerular filtration rates (GFR) and effective renal plasma flows (ERPF), were determined for up to 24 weeks after irradiation. From these data dose-effect curves were obtained by determining, at each dose level, the percentage of irradiated kidneys which exhibited a greater than or equal to 50 per cent reduction in GFR and ERPF. 60Co gamma-ray doses were normalised to approximately equivalent X-ray doses. The dose-effect curves were fitted by probit analysis, and ED50 values (+/- SE) calculated for both GFR and ERPF. Similar conclusions could be drawn from the results obtained following the determination of both GFR and ERPF. However, the ED50 values obtained for ERPF were significantly lower than those for GFR (p less than 0.05). In 14-week-old animals the kidney of UI animals was significantly more radiosensitive than that of BI animals, i.e. the ED50 values, in terms of GFR, were 8.74 +/- 0.31 Gy and 10.97 +/- 0.23 Gy, respectively (p less than 0.001). In 45-week-old pigs the reverse was true; the ED50 values were 12.67 +/- 0.34 Gy and 8.78 +/- 0.15 Gy (p less than 0.001) for unilateral- or bilateral-irradiated animals, respectively. The kidney of BI mature animals appeared to be as radiosensitive as the UI immature pigs. Thus the renal response to radiation was markedly influenced by the age of the animals and by the proportion of the renal tissue irradiated. PMID- 2569014 TI - Inhibition of DNA replication by ionizing radiation is mediated by a trans-acting factor. AB - Gamma irradiation of a human cell line containing an extrachromosomal plasmid results in inhibition of the replication of both genomic and plasmid DNA. This inhibition is observed at doses of radiation which produce an insignificant amount of damage in the plasmid DNA molecules. These results indicate that radiation-induced inhibition of DNA replication is mediated by a trans-acting factor. PMID- 2569015 TI - Inhibition and recovery of semiconservative DNA synthesis in normal and solar UV sensitive ICR 2A frog cell lines following the induction of non-dimer DNA damage by sunlamp UV greater than 315 nm. AB - Cultures of the solar UV-sensitive cell lines, DRP 36 and DRP 153, and of the parental ICR 2A cell line, were exposed to 150 kJ/m2 of sunlamp UV greater than 315 nm plus photoreactivating light. This treatment resulted in the induction primarily of non-dimer DNA damage. Following either a 0, 3, 6, 12 or 24 h incubation, the cultures were pulse-labelled with [3H]thymidine, and the synthesis of different size classes of replicon intermediates measured using the alkaline step elution assay. For all three cell lines tested, an immediate depression of low molecular weight DNA synthesis was observed. This was followed by an inhibition of all size classes of replicon intermediates. Within 12 h following irradiation, recovery of DNA synthesis was observed, which was generally most apparent for low molecular weight DNA. The ICR 2A cells exhibited a nearly full recovery in all size classes of DNA synthesized by 24 h. However, a much smaller recovery of DNA synthesis was detected for the DRP 36 and DRP 153 cultures. This continued inhibition was primarily in the synthesis of full replicon size DNA, and was most pronounced for the DRP 36 cells. Hence, it appears that replicon chain elongation continues to be inhibited in these solar UV-sensitive cell lines long after irradiation. PMID- 2569018 TI - The poor statistical properties of the Fe-plot. AB - The poor statistical properties of the Fe-plot as a method of estimating the alpha/beta ratio in the linear quadratic model for multifraction data are summarized. A Monte Carlo simulation of quantal response data shows that the Fe plot method of analysis is about half as efficient as logistic regression, indicating that twice the amount of data would be needed to obtain the same accuracy in the estimate of alpha/beta if the Fe-plot was used rather than logistic regression. In addition, the confidence intervals from the Fe-plot are inappropriately too narrow and do not have the correct coverage rate. Model-based statistical methods which directly link the treatment variables, dose and number of fractions to the response are advocated. PMID- 2569016 TI - The effects of tritium on embryo development: the embryotoxic effects of [3H]tryptophan. AB - The effects of tritiated amino acids on the development of the early mouse embryo in vitro are described. [3H]tryptophan was extremely toxic, compared with [3H]glutamic acid, and inhibited growth of the two-cell embryo to the blastocyst stage. The LD50 values for the two amino acids were, respectively, 1 and 296 kBq/ml of medium. The two-cell embryo was more sensitive to the irradiation from incorporated [3H]tryptophan is selectively incorporated into two chromosomal proteins in the two-cell embryo, and that those proteins were stage-specific markers of development. PMID- 2569017 TI - The effect of 90Sr given to pregnant mice on spermatogenesis in the male offspring: a comparison with the effect on the ovaries in the female offspring. AB - In previous studies in which the effect of intra-uterine exposure to 90Sr on the mouse was ascertained, the foetal ovary was found to be very sensitive. This paper reports the effects of the administration of 90Sr on spermatogenesis in the male littermates of these female mice. Doses of 370 and 740 kBq 90Sr administered to the dam at day 19 of gestation were found to cause a transient retardation in the appearance of more advanced types of germ cells in the testis. In comparison with the control testes, in the 90Sr treated mice less tubular cross-sections showed spermatocytes or round spermatids at days 14 and 2i p.p., respectively. After 56 days, spermatogenesis in the 90Sr treated mice was similar to that in control mice. Doses of 92.5 and 185 kBq 90Sr had no visible effect. It is concluded that the foetal testis is much less vulnerable to the damaging effects of 90Sr administration than the ovary, in which a dose of 92.5 kBq permanently decreased germ cell numbers to 40 per cent of the control numbers. Nevertheless, presumably also in the foetal testis, many germ cell will be killed by the radiation from the 90Sr. However, in the testis the surviving stem cells apparently quickly restore their numbers at the cost of a delay in the production of differentiating cells. PMID- 2569019 TI - Effect of inhaled 239PuO2 on alveolar type II cells. AB - Morphological changes of rat alveolar type II (AT-II) cells were studied at 8 and 10 months following inhalation of 239PuO2 to elucidate the biological role of AT II cells in the induction of lung tumours. TEM micrographs of random sections of lung were analysed qualitatively and quantitatively using an automatic image analyser. Eighteen morphometric parameters were obtained according to stereological principles. The results showed that, following the inhalation of 239PuO2, AT-II cells became less differentiated and the metabolism of the pulmonary surfactant in AT-II cells was disturbed. PMID- 2569020 TI - The relationship of the resonant recognition model to effects of low-intensity light on cell growth. AB - The resonant recognition model (RRM) is a model of protein-protein and protein DNA interaction based on a significant correlation between spectra of numerical presentation of the amino acid or nucleotide sequences and their biological activity. Having compared absorption characteristics of photosensitive proteins with their RRM spectral characteristics we obtained a linear correlation between the RRM frequency space and real frequency space, which determines a scaling factor between these two frequency spaces. On applying the RRM model to several groups of peptide growth factors, characteristic RRM frequencies were revealed and the corresponding real characteristic frequencies for these groups of growth factors were calculated using the scaling factor previously obtained. The real frequency characteristics of growth factors obtained in this way correspond with maxima in the action spectra of low-intensity light irradiation effects on cell proliferation. PMID- 2569021 TI - Depression of hyperthermic potentiation in cell killing of ultraviolet light irradiated Dictyostelium discoideum by pre-heat treatment. AB - Heat treatment of 30 degrees C for 15 min immediately after ultraviolet (UV) irradiation enhanced cell killing of Dictyostelium discoideum. However, when the cells were heated at 30 degrees C for 15 min and then cultured for more than 10 min at 23 degrees C prior to UV exposure, the hyperthermic potentiation in cell killing was depressed. When these preheated cells were cultured in the presence of cycloheximide, the depression of the hyperthermic potentiation disappeared. These results suggest that the depression in hyperthermic potentiation may be the result of the induction of some heat-shock-type proteins. PMID- 2569022 TI - Effects of adrenergic agents on alpha-chymotrypsin-induced ocular hypertension in albino and pigmented rabbits: a comparative study. AB - Alpha-chymotrypsin-induced ocular hypertension in albino rabbits is widely used as an experimental model to screen potential antiglaucoma drugs. The present study compares the intraocular pressure (IOP) response following the ocular application of single or repeated adrenergic agents in conscious albino and pigmented rabbits. A single instillation of clonidine was not as effective in lowering the IOP in pigmented hypertensive rabbit eyes as in albino hypertensive eyes. Similarly, betaxolol moderately lowered the IOP in albino rabbits but induced a slight response when pigmented rabbits were used as an experimental model. Twice-a-day applications of betaxolol in pigmented hypertensive eyes permitted an identical level of IOP decrease to be reached, as observed in a one day study in albino rabbits, after at least 6 days of treatment. It has been suggested that the pigmented layers of the iris-ciliary body may act as sites for topically applied antiglaucoma drugs. Non-specific binding could explain in part the frequent discrepancy observed between the preclinical results obtained in albino hypertensive rabbit eyes and clinical results obtained in glaucomatous human eyes. PMID- 2569024 TI - Effects of islet hormones on insulin secretion from cloned B cell lines, HIT-T15 and RINm5F. AB - The effects of the islet cell hormones glucagon, somatostatin-28 and pancreatic polypeptide on insulin secretion from cultured cloned pancreatic B cells (HIT-T15 and RINm5F) have been investigated. Glucagon stimulates the secretion of insulin from HIT-T15 cells in the absence and presence of glucose and from RINm5F cells in the absence and presence of glyceraldehyde. HIT-T15 cells were more sensitive to the stimulatory effect of glucagon than RINm5F cells. Somatostatin-28 and pancreatic polypeptide, both alone and in combination, reduced glucose- and glucagon-stimulated insulin release from HIT-T15 cells and glyceraldehyde- and glucagon-stimulated insulin release from RINm5F cells. HIT-T15 cells were more sensitive to the inhibitory actions of somatostatin-28 and pancreatic polypeptide than RINm5F cells. This study supports the hypothesis that insulin release from normal B cells may be modified by the paracrine activity of islet hormones, glucagon, somatostatin and pancreatic polypeptide and probably occurs before any fine tuning imposed by subsequently released insulin. PMID- 2569023 TI - Molecular defects in the insulin receptor in patients with leprechaunism and in their parents. AB - Leprechaunism is a genetic form of insulin resistance characterized by severe growth retardation and early death. To clarify the molecular basis of the insulin resistance, we investigated the insulin binding and kinase properties of the insulin receptor and the receptor gene in cultured skin fibroblasts of two patients (Ark-1 and Ark-2) with leprechaunism and in those of three of their parents. Specific insulin binding to fibroblasts was markedly reduced (less than 25% of control) in both patients with leprechaunism but was essentially normal in the parents. In contrast, insulin receptor autophosphorylation in 1% Triton X-100 cell lysates was reduced in both patients and parents. In Ark-1, the 70% reduction in autophosphorylation correlated with the decrease in binding, whereas in Ark-2 and in the three parents included in the study, autophosphorylation of the insulin receptor was reduced below the level accounted for by a change in receptor content. Analysis of the insulin receptor gene by hybridization with the receptor cDNA probes revealed no gross defect in either Ark-1 or Ark-2. Both parents of Ark-2 were heterozygous for a restriction fragment length polymorphism in the beta-subunit detected with Bam HI digestion (observed in 15% of controls). Ark-2 was homozygous for the more common allele of this polymorphism (observed in 84% of controls). Thus, we have biochemically characterized a new family of leprechaunism (Ark-2) and have found insulin receptor phosphorylation defects in their phenotypically normal parents. PMID- 2569025 TI - The importance of lipid solubility and receptor selectivity of beta-adrenoceptor blocking drugs for the occurrence of symptoms and side-effects in out-patients. AB - Short-term studies indicate that the side-effects of adrenergic beta-receptor blockade may be influenced by lipid solubility and receptor selectivity of the blockers. The aim of the present study was to investigate the extent to which lipid solubility and receptor selectivity are related to the occurrence of symptoms during ordinary beta-blocker treatment in out-patients. Two hundred and five patients (mean age 60 years) on beta-blockade were interviewed about their symptoms. Eighty-five per cent of the patients had been on beta-blocker treatment for more than 1 year. The blockers most commonly used were atenolol (51.0%), timolol (20.0%), propranolol (18.5%) and metoprolol (7.8%). beta-Blocker treatment was associated with a significantly increased frequency of central nervous system and peripheral symptoms. The increase in central nervous system symptoms was significantly smaller in those on beta-blockers with low rather than with intermediate (P = 0.015) or high lipid solubility (P = 0.007). The increase in peripheral symptoms was not significantly different with beta 1-selective and non-selective blockers. The results indicate that the liability of beta-blockers to cause side-effects on the central nervous system is determined partly by their lipid solubility, and that receptor selectivity has only a small impact on their tendency to cause peripheral side-effects. PMID- 2569027 TI - A novel subset of CD2-, CD3/T cell receptor alpha/beta+ human peripheral blood T cells. Phenotypic and functional characterization of interleukin 2-dependent CD2 CD3+ T cell clones. AB - It is generally believed that CD2 (T11, sheep erythrocyte receptor) is expressed on all human T cells. In the present study we have identified and characterized a minor subset of CD2- CD3/TCR alpha/beta+ T cells in the peripheral blood of healthy individuals. CD2-CD3+ T cells were enriched in PBMC depleted of plastic adherent macrophages, E-rosetting (i.e., CD2+) T cells and surface Ig+ B cells. CD2-CD3+ T cells accounted for 0.1-0.8% of PBMC in six individuals. IL-2 dependent long-term clones of CD2-CD3+ T cells neither reacted with a panel of anti-CD2 mAbs nor expressed detectable levels of CD2 mRNA by Northern blot analysis. These clones, however, expressed a full-length TCR C beta mRNA and reacted with mAbs against TCR-alpha/beta, CD3, and CD4, and thus were mature T cells. CD2-CD3/TCR+ T cell clones could be triggered into proliferation, IL-2 production, and cytotoxic effector activity by anti-CD3 and anti-TCR mAbs. We conclude that (a) a minor subset of CD2-, CD3/TCR-alpha/beta+ T cells is present in normal peripheral blood; and (b) expression of CD2 at the level of protein and/or mRNA is not required for T cell signaling via the CD3/TCR molecular complex. PMID- 2569026 TI - Leukocyte function-associated antigen 1 is an activation molecule for human T cells. AB - The leukocyte function-associated antigen 1 (LFA-1) molecule is well established as a surface protein involved in cellular adhesion and interaction, but there has been little information about whether engagement of this molecule can also directly modify cellular activation. These studies demonstrate that crosslinking the LFA-1 molecule on human T cell clones transmits a unique signal to the cell. Crosslinking LFA-1 alone did not increase intracellular calcium ([ CA2+]i), nor did crosslinking LFA-1 activate the cells as measured by IL-2 production or [3H]thymidine incorporation. However, when CD3 and LFA-1 were crosslinked, a more prolonged calcium signal was observed than when CD3 alone was crosslinked. Moreover, IL-2 production and DNA synthesis were greatly augmented. These responses could be demonstrated when LFA-1 was crosslinked via either the alpha or the beta chain, and required surface expression of the LFA-1 molecule as no enhancement was observed in T cell clones from a child with leukocyte adhesion deficiency. The enhancement of cellular activation by LFA-1 did not require that it be directly crosslinked to the CD3 complex. Thus, crosslinking LFA-1 alone with isotype-specific secondary antibodies on cells also pretreated with an anti CD3 mAb of a different Ig isotype stimulated the cells as effectively as crosslinking both surface antigens with GaMIg. Similarly, a delayed, but sustained increase in [Ca2+]i was elicited. This increase in [Ca2+]i and the enhanced functional responses required engagement of CD3 with an intact bivalent anti-CD3 mAb, as crosslinking LFA-1 on cells also reacted with Fab fragments of an anti-CD3 mAb did not increase [Ca2+]i, nor activate the cells. These data indicate that LFA-1 can convey activation signals to T cells. Synergism in signaling can be observed upon crosslinking of LFA-1 and independently crosslinking CD3. In the physiologic interaction between T cells and accessory cells, the interaction of LFA-1 with its ligand, intercellular adhesion molecule 1, may therefore not only facilitate cellular adhesion, but also may amplify T cell activation by delivering costimulatory signals. PMID- 2569028 TI - Antidepressants given repeatedly increase the behavioural effect of dopamine D-2 agonist. AB - The effects of single or repeated doses of antidepressant drugs (imipramine, amitriptyline, citalopram, mianserin) on rat locomotor hyperactivity induced by quinpirole, a dopamine D-2 receptor agonist, was investigated. Single doses of antidepressants do not change the effect of quinpirole, but enhance it when they are administered repeatedly. This enhancement is inhibited by (+/-)-sulpiride, a dopamine D-2 receptor antagonist. The results obtained indicate that the enhancement of dopaminergically-stimulated hyperactivity induced by repeated doses of antidepressants is mediated by dopamine D-2 receptors. PMID- 2569029 TI - Effects of intrastriatal blockade of glutamatergic transmission on the acquisition of T-maze and radial maze tasks. AB - Prefrontal cortex and neostriatum constituting the prefrontal system are connected by glutamatergic neurones. The involvement of this corticostriatal projection in control of maze performance of rats was investigated. Glutamatergic transmission mediated by N-methyl-D-aspartate (NMDA) receptors was blocked by intrastriatal injections of dl-2-amino-5-phosphonovaleric acid (AP-5) (50 nmole in 0.5 microliters). In experiment 1, intrastriatal AP-5 was found to increase the number of errors during acquisition of a delayed alternation task in a T maze. In experiment 2, the effect of intrastriatal AP-5 on acquisition of different 8 arm maze tasks was investigated. AP-5 did not affect the number of reentries on spontaneous and reinforced alternation; pre- and postdelay errors on delayed alternation were not altered. Therefore, intrastriatal NMDA receptor blockade impairs acquisition of a delayed alternation in a T-maze, while intrastriatal blockade of NMDA receptors does not affect acquisition of different 8 arm maze tasks. The impairment in the T-maze task appears not to be due to deficient acquisition of spatial information per se, since 8 arm maze performance is intact. Instead, repeated delays in the T-maze task seem to be the critical component that gives difficulties in acquisition. These difficulties in bridging successive temporal discontiguities were attributed to an increased susceptibility to external and internal interfering stimuli during delays. Thus, striatal NMDA receptors within the prefrontal system may be involved in correct response retention over the duration of delays. PMID- 2569030 TI - Primary afferent responses of a crustacean mechanoreceptor are modulated by proctolin, octopamine, and serotonin. AB - Modulation of sensory responses recorded intracellularly in primary sensory afferents of a crustacean proprioceptor is described. The neuropeptide proctolin enhances the sensory response, whereas the bioamines octopamine and serotonin depress it. The lobster oval organ of the second maxilla, a simple stretch receptor lacking centrifugal control, provides a useful model for studies on nonsynaptic modulation at peripheral sensory loci. Its three large afferents, X, Y, and Z, were prepared for intracellular recording and tested under five experimental conditions: (1) when fully rested, (2) when adapted to maintained stretch and firing tonically, (3) when showing reduced responses after habituation to repetitive stimulation, (4) not stretched but depolarized with current injections, (5) after TTX blockade. The results, taken together, indicate that conductances contributing to the overall amplitude of the receptor potential are major targets for modulators. Thus proctolin increased receptor potential amplitudes with consequent augmentation of spiking, whereas serotonin and octopamine depressed the receptor potentials, often to subthreshold levels with loss of spiking. Octopamine was a less potent agent than serotonin and failed to act upon fibers under TTX blockade. Fibers Y and Z consistently showed sensitivity to the modulators tested. The largest fiber, X, typically was resistant to proctolin, octopamine, and serotonin. Threshold concentrations of 10(-10)-10(-11) M determined in vitro are well below the circulating levels for serotonin and octopamine found in vivo. Proctolin, however, is usually not detectable in the hemolymph, and it is suggested that a significant site of proctolin release may be the oval organ itself. PMID- 2569031 TI - Genetic factors in sleep disorders. AB - Several sleep disorders have a genetic basis. These conditions include the narcoleptic syndrome, sleep walking, periodic movements in sleep, circadian delay syndromes and familial insomnia. These disorders illustrate different control mechanisms involved in sleep and wakefulness, including those determining the prevalence and timing of NREM and REM activity, somatomotor inhibition and excitation, autonomic discharge, and the circadian framework of sleep. The genetic defect in narcolepsy has been localised to the short arm of chromosome 6, but the chromosomal localisations of the genetic basis for the other disorders are not known. Also, with the possible exception of acetylcholine, no definite neurotransmitter involved in any aspect of sleep regulation has been positively identified and the biochemical defect in narcolepsy is not known. PMID- 2569032 TI - HER-2/neu oncogene protein and prognosis in breast cancer. AB - Amplification of the HER-2/neu oncogene was recently reported to predict poor clinical outcome in node-positive breast cancer patients. Since expression of the oncogene as its protein product might be even more closely related than gene amplification to disease progression, we have now examined levels of the HER 2/neu oncogene protein for its prognostic potential in both node-positive and node-negative breast cancer. Using Western blot analysis, levels of this protein were determined in 728 primary human breast tumor specimens. We examined relationships between this protein and other established markers of prognosis, as well as clinical outcome. In node-negative patients (n = 378), the HER-2/neu protein failed to predict disease outcome. However, in node-positive patients (n = 350), those patients with higher HER-2/neu protein had statistically shorter disease-free (P = .0014) and overall survival (P less than .0001) than patients with lower levels of the protein. Higher HER-2/neu protein was found in tumors without estrogen receptor (ER) (P = .02) or progesterone receptor (PgR) (P = .0003), and in patients with more than three positive lymph nodes (P = .04). A significant correlation between levels of the HER-2/neu gene protein and amplification of the gene itself was also found (n = 48, P less than .001). Multivariate analyses in these patients showed that the HER-2/neu protein is a significant independent predictor of both the disease-free and the overall survival in node-positive breast cancer, even when other prognostic factors are considered. PMID- 2569033 TI - Electrophoretic analysis of 248 clinical breast cancer specimens for P glycoprotein overexpression or gene amplification. AB - Multiple drug resistance (MDR), consisting of acquired cross resistance to anthracyclines, vinca alkyloids, and other antineoplastic antibiotics, has been described in a variety of cell lines. This MDR phenotype is associated with overexpression and sometimes amplification of a gene coding for a 170 kDa glycoprotein, termed P-glycoprotein. To understand the role of this mechanism in clinical breast cancer, 248 breast cancer specimens representing both untreated primary and refractory relapsing disease were probed for evidence of P glycoprotein gene amplification or overexpression using Southern, Northern, or Western blot techniques. In no case was an increase in P-glycoprotein gene copy number or expression detected. Though these findings do not necessarily rule out a role for P-glycoprotein in mediating drug resistance in breast cancer, electrophoretic analysis of clinical specimens is unlikely to provide useful predictive information. More sensitive assays must be developed to overcome the difficulties inherent in analyzing heterogenous tissue samples. PMID- 2569034 TI - Characteristics and distribution of receptors for [D-TRP6]-luteinizing hormone releasing hormone, somatostatin, epidermal growth factor, and sex steroids in 500 biopsy samples of human breast cancer. AB - Binding capacities and apparent dissociation constants of receptors for [D-Trp6] luteinizing hormone-releasing hormone [( D-Trp6]-LH-RH), somatostatin (SS-14), epidermal growth factor (EGF), and estrogen and progesterone were determined in 500 breast cancer specimens using multipoint assays. Specific binding sites greater than 10 fmol/mg cytosol protein for estrogen were found in 408 carcinomas (81.6%), and for progesterone in 340 specimens (68%). High affinity EGF receptors were present in membrane preparations from 335 samples (67%). In 260 of 500 samples (52%), two classes of [D-Trp6]-LH-RH membrane receptor sites were also detected, one class showing high affinity and low capacity, and the other class showing low affinity and high capacity; 178 biopsy samples (35.6%) exhibited binding sites for SS-14. Statistically significant inverse correlations were found between the binding capacities of estrogen and EGF receptors as well as between Bmax of progesterone and EGF receptors. Significant positive correlations were demonstrated between binding capacities of estrogen and progesterone and between Bmax of high affinity and low affinity binding sites of [D-Trp6]-LH-RH receptors. However, no correlation was found between the dissociation constants of different receptor sites in human breast cancer specimens. These results demonstrate that numerous human breast cancers, in addition to receptors for estrogen and progesterone, also show binding sites for EGF, [D-Trp6]-LH-RH and SS 14. The methods described herein permit a routine quantification of receptor sites for [D-Trp6]-LH-RH, SS-14, and EGF in membrane preparations of biopsy samples of breast cancer and can be used in conjunction with the determination of estrogen and progesterone receptors in nuclear-cytosolic extracts. The simultaneous measurements using a microanalytic approach allow the determination of peptide and steroid hormone receptors that might be involved in the response mechanisms of human breast cancer. It should be possible to correlate the levels of these receptors with clinical parameters to better identify endocrine responsive neoplasms. This approach might be useful to guide a rational hormonal therapy in women with breast cancer. PMID- 2569035 TI - An immunohistochemical and in situ hybridization study of c-myc and c-erbB-2 expression in primary human breast carcinomas. AB - In previous studies of the expression and organization of proto-oncogenes in human breast a significant correlation has been found between amplification of c myc and c-erbB-2 genes in carcinomas and poor short-term prognosis. Gene expression was estimated by analysis of total RNA from tissues, and similarly assessment of gene organization relied upon extraction of DNA from tissues. The present study has compared the expression of c-myc and c-erbB-2 mRNA as determined by in situ hybridization, and c-myc and c-erbB-2 protein expression detected by immunohistochemistry in a group of carcinomas for which there was knowledge of genomic organization and/or expression. Formalin-fixed, paraffin embedded tissues of 38 carcinomas were assessed for the presence of c-myc protein, and 13 of these were examined for c-myc mRNA by in situ hybridization. Similarly processed tissue from 14 tumours was tested for c-erbB-2 protein using the antiserum 21N and ten of these carcinomas studied for c-erbB-2 mRNA localization. There was a good correlation between gene amplification, the presence of c-erbB-2 protein and mRNA: both the latter were detected in six of the seven carcinomas with an amplification but in none without. For some carcinomas there was a good correlation between c-myc protein and mRNA levels. Three carcinomas with gene amplification had a lower percentage of cells with detectable protein than showed hybridization for mRNA. Other carcinomas had a lower level of mRNA expression than protein. Neither approach could predict which carcinomas had amplification of the c-myc gene.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569036 TI - Prediction of later development of thyrotoxicosis or central hypothyroidism from the cord serum thyroid-stimulating hormone level in neonates born to mothers with Graves disease. PMID- 2569038 TI - Changes of testicular cholesteryl ester hydrolase activity in experimentally cryptorchid rats. AB - A simple and reliable method was developed to determine the neutral cholesteryl ester hydrolase (CEH) activity in rat testes, using cholesteryl-[1-14C]-oleate as substrate. The activity was due to a soluble enzyme present in the cytoplasm of predominantly Sertoli cells, which could be shown after depleting the testes of Leydig cells with ethane dimethyl sulphonate. This treatment also revealed that the loss of CEH activity in abdominal testes of experimentally cryptorchid rats takes place in the Sertoli cells. In prepubertal rats made unilaterally cryptorchid at birth, the CEH activity was significantly higher in the abdominal than in the scrotal testes at 16 days of age. This is earlier than any previously described biochemical change and coincides with, or may even precede, the earliest morphological changes which are accumulation of lipid droplets in the Sertoli cells. The testicular CEH activity then decreased to 30 days of age in the abdominal testes, whereas the activity increased in the contralateral, scrotal testes. When adult rats were made unilaterally cryptorchid for 24 h, the CEH activity decreased rapidly in the abdominal testes. These results suggest that a derangement in cholesteryl ester metabolism is an early event in the pathogenesis of testicular degeneration in cryptorchidism. PMID- 2569037 TI - P300 abnormality in schizophrenic subtypes. AB - P300 and other long latency event-related potentials were recorded in 65 schizophrenic subjects and results were compared to findings in 119 healthy controls. Highly significant differences in P300 latency and amplitude were found between the two groups. Forty six per cent of schizophrenics had P300 latency more than two standard deviations longer than the mean for controls, 35% had a P300 amplitude smaller than the mean for controls by the same amount, and 24% were more than two standard deviations outside the mean for controls on both measures. These differences were independent of chronicity of illness, clinical subtype, family psychiatric history or the effects of neuroleptic medication. They confirm that P300 abnormality is present as a stable trait in a high proportion of schizophrenics. The status of abnormal P300 as a biological vulnerability factor for major mental illness is discussed. PMID- 2569039 TI - Distribution of gamma-glutamyl transpeptidase in the mouse epididymis and its response to acivicin. AB - gamma-Glutamyl transpeptidase (gamma-GT), its substrate (GSH) and hydrolytic product (L-glutamic acid) were measured biochemically in mouse reproductive tissues. The epididymal caput and seminal vesicles showed the highest specific activities of gamma-GT, while GSH and L-glutamic acid were widely distributed in all tissues. Histochemically, gamma-GT displayed a strong apical and supranuclear reaction and a moderate basal activity in the ductuli efferents, a weak luminal reaction in the first, a moderate apical reaction in the second and a strong apical and supranuclear reaction in the third segment of the epididymal caput. In the epididymal corpus and cauda, the gamma-GT reaction was confined to the tubular lumina but an apical reaction was also present in the cauda. The daily administration of acivicin (12 mg/kg body weight), an irreversible inhibitor of gamma-GT, for 14 days resulted in a 60% suppression of the enzyme activity in the epididymal caput, while the gamma-GT inhibition in the kidney was greater than 95%. The treatment caused no change in the activity of alanyl aminopeptidase. Histochemically, the basal and supranuclear gamma-GT activities in the ductuli efferents and the third epididymal segment were suppressed, but the apical reactions were maintained. The in-vivo suppression of epididymal gamma-GT activity may have implications in the control of post-testicular sperm maturation. PMID- 2569040 TI - Prescribing of benzodiazepines by casualty officers. AB - The prescribing of benzodiazepines by casualty officers in a busy district hospital over a three month period was examined by a retrospective review of case notes. Benzodiazepines, mainly diazepam, were given to 1.1% of attenders, the majority of whom had disorders involving minor muscle spasm. The efficacy of diazepam in these conditions, as well as its potential for dependence, is discussed. PMID- 2569041 TI - 5-HT1A-receptor antagonism: N-alkyl derivatives of (R)-(-)-8,11 dimethoxynoraporphine. AB - Prompted by previous findings that a p-dimethoxy substitution pattern on an aromatic ring permits retention of dopaminergic agonist effects in certain ring systems, catechol derivatives of which are potent dopaminergic agonists, an 8,11 dimethoxy substitution pattern was introduced into the aporphine ring in place of the 10,11-dihydroxy moiety in apomorphine. Acid-catalyzed rearrangement of an appropriate morphine derivative provided the aporphine ring system with retention of the stereochemical integrity of the 6a asymmetric center. The hydroxyl group at position 10 was removed by catalytic hydrogenolysis of its phenyltetrazoyl ether. The methyl ether of the resulting 11-hydroxyaporphine was iodinated in high yield at position 8 with trifluoroacetyl hypiodite. This is the first account of synthesis of an iodinated aporphine derivative. The 8-iodo substituent was replaced with methoxyl by reaction with sodium methoxide and cuprous iodide. Neither the N-methyl target compound 7 nor the N-n-propyl derivative 8 demonstrated dopaminergic nor serotonergic agonism. However, 7 exhibited receptor binding characteristics and other pharmacological properties suggesting that it may be a 5-HT1A receptor antagonist. PMID- 2569042 TI - Synthesis and in vitro pharmacology of arpromidine and related phenyl(pyridylalkyl)guanidines, a potential new class of positive inotropic drugs. AB - Replacement of the cimetidine moiety in impromidine (1,N1-[3-(1H-imidazol-4 yl)propyl]-N2-[2-[[(5-methyl-1H-imidazol-4- yl)methyl]thio]ethyl]guanidine) by more lipophilic H2-nonspecific pheniramine-like structures resulted in potent H2 agonists with up to 160 times the activity of histamine in the isolated, spontaneously beating guinea pig right atrium. Additionally, the compounds proved to be moderate H1 antagonists. Highest H2-agonistic potency was found in compounds characterized by a three-membered carbon chain connecting the aromatic rings and the guanidine group. The activity in the atrium was increased 2-4-fold by halogen substituents in the meta or para position of the phenyl ring. Highest H1-antagonistic potency resides in the group of para-halogenated compounds, p-F representing the optimal substituent in both receptor models. The corresponding guanidine 52 (arpromidine, N1-[3-(4-fluorophenyl)-3-pyridin-2-ylpropyl]-N2-[3-(1H imidazol-4- yl)propyl]guanidine) combines about 100 times the activity of histamine at the H2 receptor with H1-antagonistic potency in the range of pheniramine. Further increase in the activity on the atrium was achieved by disubstitution with halogen on the phenyl ring, such as 3,4-F2, 3,5-F2, and 3,4 Cl2 (63-65). The 2-pyridyl group in arpromidine was replaced by 3-pyridyl without significant change in H2 agonistic activity, whereas the 4-pyridyl and phenyl analogues were less active. The rank order of potency in the atrium was in good agreement with the positive inotropic effects found in isolated, perfused guinea pig hearts, where 63-65 were the most potent compounds as well. PMID- 2569043 TI - Retention of chromosome 3 in extrapulmonary small cell cancer shown by molecular and cytogenetic studies. AB - In small cell lung carcinoma, one of the short arms of chromosome 3 is typically lost. To investigate chromosome 3 in extrapulmonary small cell carcinoma, we used DNA probes that detect restriction-fragment-length polymorphisms at loci on 3p. These probes were used to study DNA extracted from tumors and normal tissues and/or tumor cell lines from five patients with extrapulmonary small cell cancer. Tumor DNA from four of the five patients with extrapulmonary small cell cancer retained heterozygosity at loci on 3p. Cytogenetic studies of the tumor cell lines established from these four patients showed retention of both short arms of chromosome 3. We conclude that the loss of genetic material from 3p observed in small cell lung cancer is not typical in extrapulmonary small cell cancer. PMID- 2569044 TI - Effect of dietary protein quality on development of aflatoxin B1-induced hepatic preneoplastic lesions. AB - The effect of the quality of dietary protein on the post-initiation development of aflatoxin B1-initiated putatively preneoplastic foci in Fischer 344 rat liver was compared with the effect of the quantity of dietary protein. Feeding wheat gluten, a low-quality protein, during the postinitiation period (between the end of aflatoxin B1 dosing and the death of the rats) inhibited the development of gamma-glutamyltransferase-positive foci when compared with that in animals fed high-quality protein (casein) diets during the same period. Lysine supplementation of wheat gluten during the postinitiation period enhanced the gamma-glutamyltransferase-positive response to a level comparable with that of the high-quality protein. These results suggest that one can inhibit the development of foci either by decreasing the quantity of protein intake and holding the quality of the protein constant or by decreasing the quality and holding the quantity constant. PMID- 2569045 TI - Isolation of a Thy-1-like glycoprotein from cat brain: distribution in retina and brain defined by monoclonal antibodies. AB - Thy-1 is a glycoprotein primarily expressed in the nervous system of a large number of vertebrate species. A concanavalin-A binding glycoprotein with a molecular weight of 25,000 daltons has been isolated from cat brain, which corresponds to the Thy-1 molecule isolated in other species. Monoclonal antibodies were generated against this cat Thy-1 molecule, and immunohistochemical staining on adult cat brain frozen sections demonstrated a localisation of the molecule to regions rich in synapses. In the cerebellum, the immunoperoxidase staining with the antibodies was intense in the molecular layer, the fibres of the granular layer, and the Purkinje cell bodies. This distribution of Thy-1 is consistent with that seen with the rodent and chicken Thy-1 molecule. In the retina, one antibody (WEHY-MS-1) detected the cat Thy-1 molecule in the ganglion cell layer and on fibres of the inner plexiform and inner nuclear layers. Another antibody (WEHY-MS-2) produced a similar retinal staining pattern except that the ganglion cell layer was devoid of Thy-1. Although the Thy-1 molecule in the rodent and chicken retina have been reported in the ganglion cell and inner plexiform layers, there have been no consistent reports of Thy-1 in the inner nuclear layer. This suggests that this putative cat Thy-1 molecule may have a retinal distribution pattern different from that seen in other species. PMID- 2569046 TI - [Restriction fragment length polymorphisms of the antithrombin III gene- examination of the normal Japanese, abnormal AT III (Toyama) and AT III deficient families]. PMID- 2569048 TI - [Treatment of chronic urticaria of unknown origin with a combination of H1 and H2 antihistaminics]. PMID- 2569047 TI - [Current problems of cardiology in the practice of an internist]. PMID- 2569049 TI - RFLPs of the phenylalanine hydroxylase gene in the Italian population. AB - Different mutations of the phenylalanine hydroxylase (PAH) gene leading to phenylketonuria (PKU) have been described associated with specific haplotypes in several European countries. In order to investigate the distribution of DNA haplotypes in Italy, restriction fragment length polymorphism (RFLP) analysis of the PAH gene was performed in nine Italian PKU patients from eight unrelated families, and in the available relatives. The analysis of eight polymorphic sites revealed haplotypes 1 and 6 in association with PKU. This pattern appears to differ from those reported for other European populations. The majority of the 14 PKU subjects studied showed compound heterozygosity for different haplotypes, as observed for other European series. RFLP analysis at the PAH locus allowed us to offer the possibility of prenatal diagnosis to six of the studied families. One prenatal diagnosis was performed and a normal fetus was diagnosed. PMID- 2569050 TI - Advances in the study of inherited metabolic disease. PMID- 2569051 TI - [Megaloblastic anemia caused by sulfasalazine in rheumatoid arthritis or ankylosing spondylitis?]. PMID- 2569052 TI - Prevalence of genetic haemochromatosis among diabetic patients. AB - Since diabetes mellitus is a frequent manifestation of haemochromatosis the prevalence of the disease was investigated in 418 patients attending a diabetic clinic. 21 (5%) patients had a persistently high serum ferritin (men, over 400 micrograms/l; women, over 300 micrograms/l) and 5 of these had transferrin saturations consistently over 55%. Idiopathic haemochromatosis was confirmed by liver biopsy in 4 patients, all of whom had a hepatic iron index greater than 2.0. The prevalence rate of previously unrecognised idiopathic haemochromatosis was thus 9.6 per 1000 (general population prevalence 1 in 250), suggesting that screening of diabetic patients for this genetic disease may be more cost effective than screening in the general population. PMID- 2569053 TI - Gastric glycerolipid as a receptor for Campylobacter pylori. AB - A species was detected in the lipid extract of human red blood cells and human and pig stomach tissue which was specifically recognised by isolates of Campylobacter pylori. The levels of this lipid were higher in human stomach antrum than in fundus and in adult compared with infant tissue samples. Chemical treatment of the purified substance suggests that it is a novel glycerolipid. PMID- 2569054 TI - Renal phosphate threshold and response to pamidronate in humoral hypercalcaemia of malignancy. AB - The effect of a single dose of pamidronate was studied in 15 patients with hypercalcaemia of malignancy. All patients were hydrated with saline 24 h before and 48 h after treatment. The dose of pamidronate was 30-90 mg intravenously, depending on corrected calcium level after hydration. Patients were retrospectively divided into two groups according to response to pamidronate. Corrected calcium fell to normal or below the upper limit of normal within a median of 3 days after treatment in 7 well controlled patients. In 8 poorly controlled patients corrected calcium failed to return to normal and in 7 of these, corrected calcium increased within 8 days of treatment. The best single indicator of a potent renal mechanism for the hypercalcaemia and also a poor response to pamidronate was a low renal phosphate threshold. Negative isotopic bone scans were also more common in poorly controlled patients. PMID- 2569057 TI - Troubled waters. PMID- 2569056 TI - Gallstones, bile acids, and the liver. PMID- 2569055 TI - Inhibitory effect of TNF alpha antibodies on synovial cell interleukin-1 production in rheumatoid arthritis. AB - The effect of tumour necrosis factor (TNF alpha) antibodies on synovial cell interleukin-1 (IL-1) production was investigated in 7 patients with rheumatoid arthritis and in 7 with osteoarthritis. Synovial cell IL-1 production was significantly reduced by anti-TNF alpha antibody in cultures from patients with rheumatoid arthritis, but antilymphotoxin antibody did not have this effect (except in 1 culture). In cultures from patients with osteoarthritis spontaneous IL-1 production was low, despite high concentrations of TNF alpha, and IL-1 production was not inhibited by anti-TNF alpha antibody. In rheumatoid arthritis, TNF alpha may be the main inducer of IL-1, and anti-TNF alpha agents may be useful in treatment. PMID- 2569058 TI - Gastrospirillum hominis. PMID- 2569059 TI - Pregnancy and glomerulonephritis. PMID- 2569060 TI - Cold at heart. PMID- 2569061 TI - The plight of modern Bushmen. AB - The nutritional status of 51 adult Bushmen in northeast Namibia, who have been forced to abandon their traditional hunter/gatherer lifestyle and merge with pastural/urbanized Hereros of Bantu origin, was assessed. Controls were 23 local medical staff and investigators. The diet was very unbalanced, consisting of little more than refined maize meal. Any money earned was usually spent on alcoholic beverages which, at such times, replaced normal food intake. Although short stature is thought to be genetic in origin (Bushmen were on average 15 cm shorter than Hereros), body mass corrected for height was severely diminished in 30% of adults, fat stores were depleted in 70%, and arm muscle was reduced in 75%. In general, women were more depleted than men. Blood tests showed low plasma vitamin C, folic acid, and vitamin A and E concentrations compared with controls. Plasma protein concentrations suggested a high frequency of chronic liver or enteric diseases. Examination of 44 hospital patients showed that 85% had pulmonary tuberculosis, and nutritional depletion was universal. The results raise serious doubts about the survival of Bushmen as an independent ethnic group. PMID- 2569063 TI - Intercontinental spread of an epidemic group A Neisseria meningitidis strain. AB - Electrophoretic enzyme typing revealed that a single group A Neisseria meningitidis clonal complex, designated III-1, was responsible for recent epidemics in Nepal, Saudi Arabia, and Chad. Epidemiological investigations and enzyme typing profiles indicated that this clone was brought from South Asia to the Middle East by Muslims making their pilgrimage (haj) to Mecca, Saudi Arabia, in 1987. Pilgrims who became group A carriers introduced this clonal group into sub-Saharan Africa on their return from the haj. The introduction of this clonal group into sub-Saharan Africa may be responsible for the current wave of epidemics affecting the region. Although the findings suggest that clonal virulence is an important factor in the development of epidemics of meningococcal disease, other factors also seem to be necessary for the development of an epidemic. PMID- 2569062 TI - Posture and central venous pressure measurement in circulatory volume depletion. AB - The effect of posture on central venous pressure (CVP) was studied in 16 patients with circulatory volume depletion before and after fluid replacement. At presentation, measurement of CVP when supine did not reflect circulatory volume depletion, with a mean (SEM) of 0.1 cm H2O (0.6), but when sat at 45 degrees the CVP showed a striking fall in all patients to -9.7 cm H2O (1.1). After fluid replacement, the CVP was 2.3 cm H2O (0.4) when supine, and -0.4 cm H2O (0.4) at 45 degrees. In the assessment of circulatory volume depletion, CVP should be measured with the patient sat at 45 degrees, if possible: measurement of CVP in a supine patient may not detect or severely underestimate circulatory volume depletion. PMID- 2569064 TI - There is no such thing as geriatric medicine, and it's here to stay. PMID- 2569065 TI - Adequacy of refugee relief rations. PMID- 2569066 TI - Protein-losing enteropathy associated with Clostridium perfringens infection. PMID- 2569067 TI - Stroke distance in acute myocardial infarction. PMID- 2569068 TI - HIV seroconversion rates in intravenous drug abusers from northern Italy. PMID- 2569069 TI - Fetoplacental passage of zidovudine. PMID- 2569070 TI - High predictivity of neurological manifestations for HIV infection in Africa. PMID- 2569072 TI - Complement receptors: another port of entry for HIV. PMID- 2569071 TI - Bronchoscopy and infection control. PMID- 2569073 TI - Response to hepatitis B vaccine in infants born to mothers with schistosomiasis. PMID- 2569074 TI - Fibrin monoclonal antibodies in diagnosis of deep venous thrombosis. PMID- 2569075 TI - Pump "run-away" causing severe hypoglycaemia. PMID- 2569076 TI - Phenytoin in pre-eclampsia. PMID- 2569077 TI - Carboxyhaemoglobin and fatal methylene chloride poisoning. PMID- 2569078 TI - Polycystic kidney disease in the fetus. PMID- 2569079 TI - Cigarette smoking and linoleic acid. PMID- 2569080 TI - Familial intractable hiccup relieved by baclofen. PMID- 2569081 TI - Suramin for treatment of adrenocortical carcinoma. PMID- 2569082 TI - Tetracyclines for dystrophic epidermolysis bullosa. PMID- 2569083 TI - Deaths after total lymphoid irradiation for multiple sclerosis. PMID- 2569084 TI - Aminophylline and essential tremor. PMID- 2569085 TI - Association of cutis verticis gyrata with fragile X syndrome and fragility of chromosome 12. PMID- 2569086 TI - Halothane and the atmosphere. PMID- 2569087 TI - Cervical screening management. PMID- 2569088 TI - Informing patients. PMID- 2569089 TI - Academic boycotts. PMID- 2569090 TI - Ask your pharmacist. PMID- 2569091 TI - Cost-benefit of self prescribing. PMID- 2569092 TI - High-dose recombinant interleukin-2 for chronic viral hepatitis. PMID- 2569093 TI - Digoxin in heart failure. PMID- 2569094 TI - Acute psychosis after mefloquine. PMID- 2569096 TI - Superoxide radical generation by inflamed human synovium after hypoxia. PMID- 2569095 TI - Who needs steroid receptor assays? PMID- 2569097 TI - Upper airways obstruction and cerebrovascular accident in children with sickle cell anaemia. PMID- 2569099 TI - Maternal pyrexia and the fetus. PMID- 2569098 TI - Erythropoietin and autologous blood donation. PMID- 2569100 TI - What must women be told about the pill? PMID- 2569101 TI - Hepatic microsomal glucose-6-phosphatase system and sudden infant death syndrome. AB - Microassay techniques and monospecific antibodies were used to study the hepatic glucose-6-phosphatase system in liver samples from 55 infants who had died suddenly and unexpectedly, including 38 victims of sudden infant death syndrome (SIDS). Raised hepatic glycogen was found in 10, all of whom had a diagnosis of SIDS, and in 1 other infant who was already known to have type 1b glycogen storage disease (deficiency of transport protein T1). Of the 10 infants with raised hepatic glycogen who had a diagnosis of SIDS, 8 had glucose-6-phosphatase deficiency (type 1a glycogen storage disease), and 2 had transport protein T2 deficiency (type 1c glycogen storage disease). PMID- 2569102 TI - Hepatitis C virus antibodies among risk groups in Spain. AB - The frequency of hepatitis C virus (HCV) infection in Spain was assessed by means of a recombinant-based immunoassay for serum anti-HCV antibodies. 836 serum samples were tested from 676 patients selected according to their risk of blood borne viral infections and presence of liver disease. Among patients at high risk of infection (with or without liver disease) anti-HCV antibodies were found in 85% of prospectively followed patients with post-transfusion non-A, non-B hepatitis, 62% of patients with chronic hepatitis or cirrhosis and a history of blood transfusion, 70% of haemophiliacs receiving replacement therapy, 70% of intravenous drug abusers, and 20% of haemodialysis patients. Only 8% of homosexual men infected with human immunodeficiency virus and 6% of female contacts of drug abusers were positive. Among patients with liver disease and no history of parenteral exposure to blood, anti-HCV antibodies were detected in 38% with cryptogenic, alcoholic, or primary biliary cirrhosis and in 44% with chronic active hepatitis. Among healthy subjects without risk factors for hepatitis the overall prevalence of anti-HCV was 1.2%. PMID- 2569103 TI - Anti-hepatitis C antibodies and non-A, non-B post-transfusion hepatitis in The Netherlands. AB - In a prospective study carried out in the Netherlands (1984-86) to establish the incidence of post-transfusion hepatitis non-A, non-B (PTH-NANB) in patients undergoing open heart surgery, 393 patients received 5315 blood product transfusions. PTH-NANB developed in 9 patients (index cases); stored serum samples from these patients and from 9 control patients, matched for age, sex, and number of blood product transfusions, as well as serum samples of all implicated blood products, were selected retrospectively. Sera were tested under code with a radioimmunoassay for the detection of antibodies to hepatitis C virus (anti-HCV). PTH-NANB patients received 151 blood product transfusions and control patients 140. 4 of 9 PTH-NANB patients (3/5 chronic, 1/4 acute resolved hepatitis) and 0/9 controls seroconverted. 7 of the transfusions given to PTH NANB patients but none of those given to control patients were anti-HCV positive. In 7 of 9 serum sets from PTH-NANB index cases plus implicated donors, either a donor or the recipient was anti-HCV positive. Among the donors implicated in transmission of PTH-NANB there was a strong correlation between raised alanine aminotransferase levels and the presence of anti-HCV antibodies. PMID- 2569104 TI - New susceptibility haplotype for type 1 diabetes. DIME Study Group. AB - In a prospective family study in Finland HLA genotyping was carried out for 1610 individuals from 422 consecutively registered families of children aged 14 years or younger with newly diagnosed insulin-dependent diabetes mellitus (IDDM). A haplotype (A2, Cw1, Bw56, w6, DR4) that has not been identified previously was the third most common (5.5%) among 746 haplotypes found in the probands. Only the haplotypes A1, Cw7, B8, w6, DR3 and A2, Cw3, Bw62, w6, DR4, which are well known to occur with high frequency in IDDM, were more frequent (10.7% and 9.7%, respectively). Among the 30 families in which a parent had IDDM, the newly identified haplotype was the most common haplotype transmitted from the diabetic parent to the proband (16.7%). Among 642 "non-diabetic" haplotypes (parental haplotypes not found in probands, or siblings or parents with IDDM), this haplotype was found only twice. This haplotype may partially explain the high incidence of IDDM in Finland. PMID- 2569106 TI - Will the real hepatitis C stand up? PMID- 2569105 TI - Adjuvant xamoterol or metoprolol in patients with malignant ventricular arrhythmia resistant to amiodarone. AB - In a randomised cross-over study, six patients with recurrent sustained ventricular tachycardia (VT) were treated with 3 regimens--amiodarone, amiodarone plus metoprolol, and amiodarone plus xamoterol. All patients had poor left ventricular function and were resistant to multiple drugs. Xamoterol (a partial beta-agonist) was more effective than metoprolol as adjuvant therapy to amiodarone in the control of recurrent sustained ventricular arrhythmias and was not associated with any clinical deterioration of ventricular function. Xamoterol was also more effective than metoprolol for suppression of VT at programmed stimulation and as effective as metoprolol for suppression of VT on exercise. Exercise tolerance was significantly greater during treatment with xamoterol/amiodarone than during treatment with metoprolol/amiodarone or with amiodarone alone. PMID- 2569107 TI - Bitter reinstatement. PMID- 2569108 TI - Clinical signs in heart failure. PMID- 2569109 TI - Diffuse Lewy body disease. PMID- 2569110 TI - Oral cancer. PMID- 2569112 TI - Autotransplants in leukaemia. AB - With bone marrow autotransplants, favourable results are reported in subjects with leukaemia in first and second remission but not in those with advanced disease. Whether autotransplants are equivalent or superior to other therapies such as chemotherapy and allogeneic transplants is uncertain, since prospective trials are not reported and data analysis is confounded by selection of subjects and time-censoring. Major problems of autotransplants include difficulty in eradicating leukaemia in the subject, lack of the graft-versus-leukaemia effect associated with allotransplants, and reinfusion of leukaemia cells. PMID- 2569111 TI - Vitamins, iron, and physical work. AB - Combined marginal deficiencies of iron, vitamin C, and B-group vitamins are widespread in developing countries. An understanding of the functional implications poses a continuing challenge, both for the development of new techniques of investigation and for the design of effective intervention programmes. Practical intervention in developing countries also needs to take account of the interactions of micronutrient status and disease susceptibility. It is becoming clear that combined deficiencies of certain vitamins, or of iron and vitamins, although less severe than those causing the lesions of classic clinical deficiency, can seriously reduce work performance. Riboflavin, particularly, has a special relation with iron economy. PMID- 2569113 TI - Autologous bone marrow transplants: different indications in Europe and North America. Advisory Committee of the International Autologous Bone Marrow Transplant Registry (ABMTR). AB - The International Autologous Bone Marrow Transplant Registry surveyed activity at 112 centres worldwide. Transplants increased from 265 in 1981 to over 1200 in 1987. Diseases most frequently treated by autotransplantation included non Hodgkin lymphoma (22%), acute myelogenous leukaemia (19%), acute lymphoblastic leukaemia (15%), Hodgkin disease (15%), and neuroblastoma (5%). There were striking differences in the use of autotransplants between North America and Europe. Autotransplants for lymphomas and solid tumours comprised 94% of North American Autotransplants but only 41% of those in Europe. In contrast, leukaemia was the indication in 59% of Europeans but only 6% of North Americans. 65% of leukaemia autotransplants in Europe were done in first remission compared with 8% in North America. There were also differences in age and types of lymphoma autotransplanted--Burkitt and lymphoblastic lymphoma in young persons in Europe, versus large-cell lymphomas in older persons in North America. PMID- 2569115 TI - Antibody to hepatitis C virus in German blood donors. PMID- 2569114 TI - Are diarrhoea control programmes likely to reduce childhood malnutrition? Observations from rural Bangladesh. AB - Growth of rural Bangladeshi children aged 6-35 months was examined in relation to the history of diarrhoea in 1772 3-month intervals. Weight gain and linear growth were lower in intervals with a history of diarrhoea than in intervals without diarrhoea. However, comparison of weight and height gains in intervals during which diarrhoea occurred at the beginning or at the end showed that after non bloody diarrhoeas children catch up and that deficits in weight gain and linear growth were no longer apparent a few weeks later. These findings suggest that the effect of diarrhoea on growth is transient and that efforts to control diarrhoea are unlikely to improve children's nutritional status in the long term. PMID- 2569116 TI - Antibodies to hepatitis C virus. PMID- 2569117 TI - Kala-azar in a Scottish child. PMID- 2569118 TI - Spinal cord compression after long-term etretinate. PMID- 2569120 TI - Serum cholesterol-lowering activity of human monocytic colony-stimulating factor. PMID- 2569119 TI - Use of flecainide in children. PMID- 2569121 TI - Long-term results of hyperbaric oxygen therapy in multiple sclerosis. PMID- 2569122 TI - HTLV-I (Philadelphia, 1969) PMID- 2569123 TI - HTLV-I in English patients. PMID- 2569124 TI - Gene amplification to detect fetal nucleated cells in pregnant women. PMID- 2569125 TI - Faecal occult blood screening for colorectal cancer. PMID- 2569126 TI - Tacrine in Alzheimer's disease. PMID- 2569127 TI - Economics for medical students. PMID- 2569128 TI - Outcome prediction in intensive care. PMID- 2569129 TI - Health inequalities in Britain and Sweden. PMID- 2569130 TI - Irish health services. PMID- 2569131 TI - Pulsed electromagnetic energy and childbirth. PMID- 2569132 TI - Aspergillus in pepper. PMID- 2569133 TI - Isolation of Weeksella zoohelcum in septicaemia. PMID- 2569134 TI - Borrelia antibody titres: course in neurological diseases. PMID- 2569135 TI - Is lead mobilised by cisplatin? PMID- 2569136 TI - Immunosuppressive properties of cyclosporin metabolites. PMID- 2569137 TI - Antihistaminic drugs that reverse chloroquine resistance in Plasmodium falciparum. PMID- 2569138 TI - Effective cervical cytological sampling. PMID- 2569139 TI - Cytomegalovirus infection and progression to AIDS. PMID- 2569140 TI - AIDS and Mycobacterium avium serovars in Sweden. PMID- 2569141 TI - Dietary restriction and methylmalonicaciduria. PMID- 2569142 TI - Familial association of Crohn's and coeliac diseases. PMID- 2569143 TI - Hantavirus nephropathy in Netherlands. PMID- 2569144 TI - Schizophrenia induced by restitution of thyroid function in a predisposed patient. PMID- 2569145 TI - Pseudoautosomal locus for the cerebral dominance gene. PMID- 2569146 TI - Abortion ruling in Quebec. PMID- 2569147 TI - Latest AIDS figures. PMID- 2569148 TI - Determination of ethane and pentane in free oxygen radical-induced lipid peroxidation. AB - It has been proposed that ethane and pentane reflect free oxygen radical-induced lipid peroxidation. However, methodological difficulties limit the use of these gases for assessment of free oxygen radical activity. In the present report we describe an improved method for the accurate analysis of picomole quantities (greater than or equal to 1 pmol) of ethane and pentane. They are first quantitatively trapped into an adsorbent and then heat-desorbed directly into a capillary column for gas chromatographic quantitation. During oxidation of linolenic (n-3) and linoleic (n-6) acid, ethane and pentane were formed, respectively. Nonstimulated granulocytes formed pentane. Upon addition of phorbol 13-myristate 12-acetate, the generation of pentane was increased by 540%. Addition of superoxide dismutase plus catalase inhibited lipid peroxidation in both a cell-free system and in isolated cells. The present method is useful in the evaluation of free oxygen radical induced damage. PMID- 2569149 TI - Modulation of eicosanoid production and cell-mediated cytotoxicity by dietary alpha-linolenic acid in BALB/c mice. AB - The effects of dietary alpha-linolenic acid (18:3n-3) on fatty acid composition, eicosanoid production, and cell-mediated cytotoxic activity of immune cells before and after challenge with virus or poly I-C from BALB/c mice were studied. Weanling BALB/c mice were fed purified diets containing either 10%-by-weight corn oil or linseed oil providing a ratio of 18:3n-3 to 18:2n-6 of 1/32 or 2/1, respectively, for 6-10 weeks. Fatty acid analysis of splenocyte phospholipids showed an appreciable increase in the percentage of n-3, and a decrease in n-6, fatty acids in splenocytes from mice fed the linseed oil diet. Splenocyte prostaglandin E and peritoneal exudate cell leukotriene C production was significantly lower in the linseed oil-fed mice. In general, cell-mediated cytotoxic activity was similar for immune cells from linseed oil and corn oil-fed mice. However, 6 days after the viral challenge, splenocyte cell-mediated cytotoxic activity was significantly higher in linseed oil mice. This higher activity was associated with nonspecific cytotoxicity rather than that of viral specific cytotoxic T-lymphocytes. Cell yields from the spleen and peritoneum were frequently significantly higher in linseed oil mice. Interactions between dietary 18:3n-3, eicosanoid production, and immune cell proliferation and/or migration are discussed. In summary, feeding mice a diet rich in 18:3n-3 elevates immune cell n-3 fatty acid content, reduces eicosanoid synthesis and, to a limited extent, enhances the cell-mediated cytotoxic response to a viral challenge. PMID- 2569151 TI - New phospholipid fatty acids from the Caribbean sponge Ectyoplasia ferox. AB - The phospholipid fatty acids from the Caribbean sponge Ectyoplasia ferox were studied. The novel fatty acids 25-methyl-5,9-heptacosadienoic (1) and 26-methyl 5,9-heptacosadienoic (2) were identified in 3.4 and 2.0% abundance, respectively, representing the longest set of delta 5,9 iso and anteiso acids yet isolated from a marine sponge. The new acid 10,13-dimethyltetradecanoic (3), the unusual acid 15-methyl-11-hexadecenoic (4) and the also novel acid 9-methyl-11-hexadecenoic (5) were also identified in E. ferox. The principal sterols isolated from E. ferox were 24-ethylcholest-5-en-3 beta-ol (46%) and 24(R)-methylcholesta-5,22 dien-3 beta-ol (14%). PMID- 2569150 TI - Dietary saturated fat level alters the competition between alpha-linolenic and linoleic acid. AB - Male weanling rats were fed semi-synthetic diets high in saturated fat (beef tallow) vs high in linoleic acid (safflower oil) with or without high levels of alpha-linolenic acid (linseed oil) for a period of 28 days. The effect of feeding these diets on cholesterol content and fatty acid composition of serum and liver lipids was examined. Feeding linseed oil with beef tallow or safflower oil had no significant effect on serum levels of cholesterol. Serum cholesterol concentration was higher in animals fed the safflower oil diet than in animals fed the beef tallow diet without linseed oil. Feeding linseed oil lowered the cholesterol content in liver tissue for all dietary treatments tested. Consumption of linseed oil reduced the arachidonic acid content with concomitant increase in linoleic acid in serum and liver lipid fractions only when fed in combination with beef tallow, but not when fed with safflower oil. Similarly, omega 3 fatty acids (18:3 omega 3, 20:5 omega 3, 22:5 omega 3, 22:6 omega 3) replaced omega 6 fatty acids (20:4 omega 6, 22:4 omega 6, 22:5 omega 6) in serum and liver lipid fractions to a greater extent when linseed oil was fed with beef tallow than with safflower oil. The results suggest that the dietary ratio of linoleic acid to saturated fatty acids or of 18:3 omega 3 to 18:2 omega 6 may be important to determine the cholesterol and arachidonic acid lowering effect of dietary alpha-linolenic acid. PMID- 2569152 TI - The nucleotide sequence of the first two genes of the CFA/I fimbrial operon of human enterotoxigenic Escherichia coli. AB - An oligonucleotide probe, derived from the N-terminal amino acid sequence of the CFA/I fimbrial subunit protein, was used to identify the gene encoding this protein within a cloned DNA fragment encoding CFA/I fimbriae. The gene (cfa b) was found and sequenced. Flanking it upstream was a gene (cfa a) encoding a protein of 206 amino acids and downstream a gene (cfa c) probably encoding an 85 kDa protein was found. This genetic organisation of the CFA/I operon differs from that of other fimbrial operons in Escherichia coli. All three proteins have signal peptides. The nucleotide sequence was analysed for homology with other sequences, secondary structure, ribosomal binding sites and possible promoter sequences. A region of dyad symmetry probably involved in the regulation of translation of the cfa c gene was found at the 5' end of this gene. A region of dyad symmetry was also observed within the cfa b gene. In front of the CFA/I operon part of insertion sequence IS2 was found. This IS2 sequence was found in a number of CFA/I plasmids, obtained from strains isolated from various geographic locations. The insertion of the IS2 element in the CFA/I operon therefore probably happened rather early during evolution of CFA/I producing Escherichia coli strains. PMID- 2569153 TI - [Evaluation of the usefulness of thin-layer chromatography and orientation tests detecting various addictive drugs in biological specimens]. AB - Abuse of medicines and alcohol pose particular problems with those carrying out professional work involving movement (drivers, crane operators etc.). Most accidents are caused by ethyl alcohol, however, other substances cannot be excluded, either. This paper has been aimed at the verification of the efficiency of thin-layer chromatography and tests applied in toxicological analyses of acute poisonings in the detection of lower concentrations of certain addictive medicines. The studies were carried out on biological material (urine) of subjects who had been given, for therapeutic purposes, increased doses of medicines. The group involved 156 patients from I Clinic of Psychiatry, Medical Academy of Lodz, and the Department of Disaccustoming and Psychiatry in Zgierz. PMID- 2569154 TI - Binding of F41 and K99 fimbriae of enterotoxigenic Escherichia coli to glycoproteins from bovine and porcine colostrum. AB - F41 and K99 fimbriae of enterotoxigenic Escherichia coli were found to bind to periodate-sensitive oligosaccharides of glycoproteins from bovine and porcine colostrum. Only a minor component of casein fractions (kappa-casein) possessed receptors for one type of fimbriae (K99). Both whey and fat globule membranes were rich in glycoproteins with receptor structures. Porcine colostrum seemed to contain a higher quantity of receptors than bovine colostrum. PMID- 2569155 TI - DNA diagnosis of cystic fibrosis--an Australian perspective. PMID- 2569156 TI - Cystic fibrosis: prenatal diagnosis and carrier detection by DNA analysis. AB - The analysis of restriction fragment length polymorphism (RFLP)s was used to detect 11 polymorphisms that are linked to cystic fibrosis in 42 Australian families with at least one child with cystic fibrosis. The data from all the families were fully informative in regard to the gene for cystic fibrosis (CF). Prenatal assessment was performed for 24 of these families: seven fetuses were assessed to be homozygous for cystic fibrosis, 13 fetuses were heterozygous and three fetuses were free of the CF gene. Of the seven pregnancies in which it was predicted that the infant would be affected by cystic fibrosis, two were continued electively; both have come to term and the infants each were shown to have cystic fibrosis at birth. Of the 17 pregnancies in which it was predicted that the infant would not be affected by cystic fibrosis, 13 have come to term and all the infants but one (who has not yet been followed-up) have been shown to be unaffected by cystic fibrosis at birth. The polymerase chain reaction has been used to amplify the CS.7 and KM.19 loci close to the CF gene. This procedure allows a polymorphic site in each locus to be analysed in a much shorter time (one or two days rather than 10 days) and allows the use of very small test samples, such as dried blood on filter paper ("Guthrie blood spots"). Our observations confirm the results of overseas studies and indicate that these techniques are eminently useful for prenatal diagnosis and the detection of carriers in the vast majority of Australian families with cystic fibrosis. PMID- 2569157 TI - Analysis of DNA probes for the prenatal diagnosis of cystic fibrosis. AB - Cystic fibrosis is a common autosomal recessive disease in white persons. Prenatal diagnosis by DNA analysis became possible in families with a child who is affected by cystic fibrosis when the probes pJ3.11, metH and metD, which are linked closely to the cystic fibrosis gene (CF) were described. The recent description of the XV-2c and KM.19 probes has improved the prenatal diagnosis of cystic fibrosis greatly. The KM.19 probe alone was informative in eight of 12 families that were studied while XV-2c was informative in eight of 12 families that were studied while XV-2c was informative in only two of the 12 families. In contrast, the use of the pJ3.11, metH and metD probes in combination allowed full diagnosis in six of the 12 families. The combined use of the CF-linked probes produced informative data for all 12 families. Therefore, in most families with at least one affected living child, the first-trimester diagnosis of cystic fibrosis is possible with fetal DNA that has been prepared from chorionic villous samples. Strong linkage disequilibrium was found with both the KM.19-PstI polymorphism and the XV-2c-TaqI polymorphism and the CF gene. PMID- 2569158 TI - Antihistamine agents: new options or just more drugs? PMID- 2569159 TI - [Circulation of California encephalitis complex viruses in the northwestern Russian plain]. PMID- 2569160 TI - [Experimental study of densovirus infection transmission in blood-sucking mosquito populations]. AB - The paper presents the results of laboratory study of the modes of entomopathogenic densonucleasis virus transmission in the population of blood sucking Culicidae mosquitoes. It was established that densovirus infection was horizontally transmitted through feeding of larvae on the bodies of desonucleasis infected insects as well as contamination of the environment by infected larvae. Thus, the death rate in populations of various mosquito species attained 25.9 70.8%. Vertical transmission of the virus alters the ratio of sexes at the stage of imago; the survived females and males transmit infection to larvae. Surface sterilization of the ova doesn't prevent larvae infection, death rate among larvae attaining 7.8%. PMID- 2569161 TI - [Therapy of obstructive respiratory tract diseases. I: Beta 2-sympathomimetic drugs and anticholinergic agents (m-cholinergic receptor blockers)]. PMID- 2569163 TI - Behavioral teratology investigation of 1-butanol in rats. AB - Two concentrations of 1-butanol (3000 and 6000 ppm) were administered by inhalation to separate groups of 15 pregnant Sprague-Dawley rats for 7 hr per day throughout gestation; 18 male rats were similarly exposed for 7 hr per day for 6 weeks, and mated to unexposed females. Litters were culled to 4 female and 4 male pups and fostered to untreated controls. From days 10-90, offspring were tested as follows: a) ascent on a wire mesh screen, b) rotorod, c) open field and photoelectrically-monitored activity, d) running wheel, e) avoidance conditioning, and f) operant conditioning. Additionally, brains from 10 offspring at 21 days of age were dissected into cerebrum, cerebellum, brainstem, and midbrain. Each sample was assayed for protein and the neurotransmitters acetylcholine, dopamine, norepinephrine, serotonin, met-enkephalin, beta endorphin, and substance P. Overall, there were few behavioral or neurochemical alterations detected in the offspring following maternal or paternal exposure to either 3000 or 6000 ppm 1-butanol. This scarcity of effects is important to risk assessment extrapolations drawn from ethanol. Based on the structural similarity of 1-butanol to ethanol and long-standing observations that toxicity to adult animals generally increases with chain length among the alcohols, significant behavioral and neurochemical deviations were predicted. The scarcity of effects from butanol needs to be accounted for in hypotheses relating toxicity to alcohol chain length and in risk assessment extrapolations from findings with ethanol. PMID- 2569164 TI - Repression of transcription mediated at a thyroid hormone response element by the v-erb-A oncogene product. AB - Several recent observations, such as the identification of the cellular homologue of the v-erb-A oncogene as a thyroid-hormone receptor, have strongly implicated nuclear oncogenes in transcriptional control mechanisms. The v-erb-A oncogene blocks the differentiation of erythroid cells, and changes the growth requirements of fibroblasts and erythroblasts. Mutations in v-erb-A protein have led to the loss of its affinity for thyroid hormones but do not affect its DNA binding ability, a property required for biological activity. We report here the identification of a novel thyroid-hormone response element (TRE) in the long terminal repeat of Moloney murine leukaemia virus that binds the c-erb-A-alpha protein. The v-erb-A protein abolishes the responsiveness of this TRE to thyroid hormone, although it has a lower affinity than the normal receptor for the TRE. The data indicate that overexpressed v-erb-A protein negatively interferes with normal transcriptional-control mechanisms, and that amino-acid substitutions have altered its DNA-binding properties. PMID- 2569165 TI - Transport proteins. Export-import family expands. PMID- 2569166 TI - The yeast STE6 gene encodes a homologue of the mammalian multidrug resistance P glycoprotein. AB - Mammalian tumours displaying multidrug resistance overexpress a plasma membrane protein (P-glycoprotein), which is encoded by the MDR1 gene and apparently functions as an energy-dependent drug efflux pump. Tissue-specific expression of MDR1 and other members of the MDR gene family has been observed in normal cells, suggesting a role for P-glycoproteins in secretion. We have isolated a gene from the yeast Saccharomyces cerevisiae that encodes a protein very similar to mammalian P-glycoproteins. Deletion of this gene resulted in sterility of MATa, but not of MAT alpha cells. Subsequent analysis revealed that the yeast P glycoprotein is the product of the STE6 gene, a locus previously shown to be required in MATa cells for production of a-factor pheromone. Our findings suggest that the STE6 protein functions to export the hydrophobic a-factor lipopeptide in a manner analogous to the efflux of hydrophobic cytotoxic drugs catalysed by the related mammalian P-glycoprotein. Thus, the evolutionarily conserved family of MDR-like genes, including the hlyB gene of Escherichia coli and the STE6 gene of S. cerevisiae, encodes components of secretory pathways distinct from the classical, signal sequence-dependent protein translocation system. PMID- 2569168 TI - gamma-Glutamyl transpeptidase in dementia. PMID- 2569169 TI - Age-related changes in multiple neurotransmitter systems in the monkey brain. AB - Age-associated changes in cholinergic, monoaminergic and amino acid neurotransmitter systems were analyzed in 14 brain regions of 23 rhesus monkeys that ranged in age from 2 to 37 years. In the frontal pole, the levels of choline acetyltransferase (ChAT) activity, the density of [3H]ketanserin (serotonin type 2) binding sites and endogenous levels of dopamine, homovanillic acid and serotonin, all expressed per milligram of protein, decreased significantly with aging. In precentral motor cortex, ChAT activity decreased; in parietal and occipital cortex, the number of [3H]ketanserin binding sites decreased while the number of Na+-independent [3H]glutamate binding sites increased with age. In the caudate nucleus, endogenous levels of norepinephrine decreased. This descriptive study indicates that the aging monkey may be a very useful model for future investigations of age-associated transmitter abnormalities similar to those that occur in humans. PMID- 2569167 TI - [Analysis of the effect of quisqualate, N-methyl-D- aspartate and several blockers of amino acid receptors on synaptic transmission in the ampullae of Lorenzini]. AB - Effects of bath-application of quisqualate (Q), N-methyl-D-aspartate (NMDA) and antagonists of NMDA-receptors: D-amino-adipate (AA), 2-amino-4-phosphonobutyrate (APB), 2-amino-5-phosphonovalerate (APV) and Mg2+ as well as acidic amino acid antagonist: D-glutamylglycine (DGG) on the synapse between the electroreceptor cells and afferent fibres were studied in the ampullae of Lorenzini. Q (threshold concentration 10(-8) M) and NMDA (threshold concentration 10(-5) M) strongly excited afferent fibres. Neither AA nor APB influenced the resting and evoked activities. APV blocked the synaptic transmission. Mg2+ (30-50 mM) blocked the responses to NMDA, while Q-induced responses were not affected, APV preferentially blocked NMDA-induced responses and in lesser degree--L-aspartate (L-ASP)-induced responses. DGG blocked the synaptic transmission. It is supposed that the synaptic receptor could represent a homogeneous receptor with different binding sites to the known agonists. PMID- 2569162 TI - Common themes in microbial pathogenicity. AB - A bacterial pathogen is a highly adapted microorganism which has the capacity to cause disease. The mechanisms used by pathogenic bacteria to cause infection and disease usually include an interactive group of virulence determinants, sometimes coregulated, which are suited for the interaction of a particular microorganism with a specific host. Because pathogens must overcome similar host barriers, common themes in microbial pathogenesis have evolved. However, these mechanisms are diverse between species and not necessarily conserved; instead, convergent evolution has developed several different mechanisms to overcome host barriers. The success of a bacterial pathogen can be measured by the degree with which it replicates after entering the host and reaching its specific niche. Successful microbial infection reflects persistence within a host and avoidance or neutralization of the specific and nonspecific defense mechanisms of the host. The degree of success of a pathogen is dependent upon the status of the host. As pathogens pass through a host, they are exposed to new environments. Highly adapted pathogenic organisms have developed biochemical sensors exquisitely designed to measure and respond to such environmental stimuli and accordingly to regulate a cascade of virulence determinants essential for life within the host. The pathogenic state is the product of dynamic selective pressures on microbial populations. PMID- 2569170 TI - Individual differences in aging: behavioral and neurobiological correlates. AB - The goal of this experiment was to determine the correlations among different behavioral and neurobiological measures in aged rats. Aged Sprague-Dawley rats were given a battery of cognitive and sensorimotor tests, followed by electrophysiological assessment of sleep and biochemical measurements of various neurotransmitter systems. The behavioral tests included the following: Activity level in an open field; short-term and long-term memory of a spatial environment as assessed by habituation: spatial navigation, discrimination reversal, and cue learning in the Morris water pool; spatial memory in a T-maze motivated by escape from water; spatial memory and reversal on the Barnes circular platform task; passive avoidance; motor skills. Sleep was assessed by electrographic cortical records. The following neurotransmitter markers were examined: Choline acetyltransferase; the density of nicotinic, benzodiazepine and glutamine receptors in the cortex and caudate nucleus; endogenous levels of norepinephrine, dopamine, and serotonin in the cortex and hippocampus. The duration of bouts of paradoxical sleep was strongly correlated with several cognitive measures and selected serotonergic markers. This finding suggests that changes in sleep patterns and brain biochemistry contribute directly to deficits in learning and memory, or that the same neurobiological defect contributes to age-related impairments in sleep and in learning and memory. PMID- 2569171 TI - A D-2 dopaminergic agonist stimulates secretion of anterior pituitary immunoreactive beta-endorphin in rats. AB - Hypothalamic dopamine neurons are known to control circulating levels of immunoreactive beta-endorphin (i beta-END) by inhibiting hormone secretion by the intermediate lobe (IL) of the pituitary gland. We examined the ability of the D-2 selective dopaminergic agonist, LY141865, to influence circulating levels of i beta-END in rats and found that in contrast to inhibiting IL secretion, LY141865 increased release of i beta-END from the anterior lobe (AL). Intraperitoneal injection of 1 mg/kg LY141865 transiently increased plasma levels of i beta-END by 7-30 min after drug treatment; plasma prolactin levels were maximally reduced within 15 min and throughout the remaining 2-hour time course of treatment. Doses of 0.3 and 1.0 mg/kg of LY141865 increased circulating i beta-END to 440 and 690%, respectively, of control levels (0.38 +/- 0.12 ng/ml, mean +/- SEM, n = 6). Lower doses of the D-2 agonist (0.01-0.1 mg/kg) failed to significantly affect plasma i beta-END. Sephadex G-50 chromatography of plasma pools revealed that virtually all of the increase due to LY141865 treatment was immunoreactivity resembling beta-lipotropin in molecular size, the principal component of AL secretion of i beta-END. Furthermore, LY141865-evoked release was blocked by pretreatment of rats with dexamethasone (50 micrograms/kg i.p., 4 h) which inhibits AL but not IL secretion of pro-opiomelanocortin-derived peptides. Stimulation of i beta-END release by LY141865 was also inhibited by the general dopamine antagonist, haloperidol, (0.1-3.0 mg/kg i.p., 2 h) and by the D-2 selective antagonist, sulpiride (100 micrograms/rat i.c.v., 4 h).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569172 TI - Studies on the conditions determining the inhibitory effect of somatostatin on adrenocorticotropin, prolactin and thyrotropin release by cultured rat pituitary cells. AB - Somatostatin (SRIH) is a physiological inhibitor of growth hormone (GH) secretion, but its role in the regulation of adrenocorticotropic hormone (ACTH), prolactin (PRL) and thyroid-stimulating hormone (TSH) release is unclear. SRIH (1 pM to 1 microM) did not affect basal and corticotropin-releasing hormone (CRH) stimulated ACTH release by normal rat pituitary cells cultured in medium with 10% fetal calf serum (FCS). In cells deprived of serum for 48 h, or preincubated with the glucocorticoid-receptor-blocking agent, RU 38486, CRH-stimulated ACTH release was significantly suppressed by 1 pM to 0.10 nM SRIH. Preincubation with 5 nM dexamethasone completely abolished this inhibitory effect of SRIH on ACTH release. PRL release by pituitary cells cultured in phenol red-free culture medium with 10% estrogen-stripped FCS showed a very low sensitivity to SRIH. Increasing concentrations of 10 and 50 pM and 1 nM estradiol made PRL release by these cells significantly less sensitive to 50 nM dopamine, whereas the sensitivity to SRIH increased to a similar extent. In all instances dopamine and SRIH together exerted additive inhibitory effects, the extent of which remained similar under all conditions. After a 2-hour incubation, thyrotropin-releasing hormone-stimulated TSH secretion was significantly suppressed by 100 nM and 1 microM SRIH only in cells cultured in medium with 10% hypothyroid serum, and not in cells cultured in medium with 10% FCS. Such a difference in the sensitivity of thyrotrophs to SRIH disappeared during longer incubation. CONCLUSIONS: (1) ACTH release by normal corticotrophs is only sensitive to SRIH in the absence of the physiological peripheral feedback regulation by glucocorticoids.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569173 TI - Behavioural and morphological changes following treatment with GM-1 ganglioside of rats with an electrolytic lesion of the substantia nigra. AB - The effects of daily treatment with GM-1 ganglioside (30 mg/kg i.p.) or saline (1 ml/kg i.p.) in rats with discrete unilateral electrolytic lesions of the dorsal substantia nigra were investigated. Treatment with GM-1 ganglioside, beginning 3 days prior to surgery and continuing for 33 days post-operatively, caused reductions in the total ipsilateral turns and peak turning rates induced by apomorphine (0.75 mg/kg s.c.) between 7 and 31 days post-operatively, without altering the time course of the effect of apomorphine. No effects of GM-1 on lesion-induced changes in synaptosomal uptake of [3H]dopamine, dopamine, serotonin in the striatum or corresponding levels of metabolites were found. Treatment with GM-1 ganglioside had no effect on the rostrocaudal extent of the electrolytic lesion, but caused morphological changes at the site of the lesion, including a reduction in the density of glial cells. Treatment with GM-1 ganglioside resulted in a relative preservation of cell bodies, staining immunocytochemically for tyrosine hydroxylase, in the substantia nigra pars compacta (expressed as a percentage of the intact side), when compared with saline-treated rats. The results showed that GM-1 ganglioside promoted a partial functional recovery from apomorphine-induced circling, which may be due in part to a reduction in the extent of damage at the lesion site. PMID- 2569174 TI - Stress-induced potentiation of morphine-induced analgesia in morphine-tolerant rats. AB - This study was designed to evaluate whether or not rats that were tolerant to the analgesic action of morphine were also tolerant to stress-induced potentiation of morphine-induced analgesia. Rats were trained to drink either solutions of morphine (0.5 mg/ml) or drug-free tap water on a limited access schedule (10 min every 6 hr). The daily intake of morphine averaged 46 mg/kg. Nontolerant and rats tolerant to morphine were tested for morphine-induced analgesia (tail-flick assay), while either unstressed or stressed (i.e. immobilized in Plexiglas cylinders). Morphine produced dose- and time-dependent increases in tail-flick latencies in all groups. Increased sensitivity to analgesia induced by morphine was evident for both nontolerant and tolerant, stressed rats, when compared to their unstressed counterparts. Stress-induced potentiation of morphine-induced analgesia was characterized by dose-related increases in the peak effect and duration of the effect. Stress potentiated the analgesic effect of morphine, comparably in nontolerant (1.7-fold) and tolerant (1.5-fold) rats. Differential tolerance to analgesia induced by morphine and to stress-induced potentiation of morphine-induced analgesia suggests that different mechanisms mediate these two effects. PMID- 2569175 TI - Idebenone and vinpocetine augment long-term potentiation in hippocampal slices in the guinea pig. AB - The effects of idebenone and vinpocetine which reportedly prevent impairment of learning and memory were studied in vitro, on the long-term potentiation of the population spike in the pyramidal layer of CA3 region of slices of hippocampus in the guinea pig. Idebenone (10(-9) M-10(-6) M) or vinpocetine (10(-7) M-10(-6) M) significantly augmented long-term potentiation in the mossy fibre-CA3 pyramidal cell system, without any significant changes in population spikes in the absence of tetanic stimulation. These results suggest that both drugs have direct actions on the hippocampal neurones to augment long-term potentiation at fairly small concentrations. Further, when the two drugs were applied together, the augmenting effects were additive. PMID- 2569176 TI - Effect of dopaminergic drugs on enhancement of the binding of quinuclidinyl benzilate by phencyclidine in vivo. AB - Phencyclidine has been shown to enhance the specific binding of [3H]quinuclidinyl benzilate (QNB) in the brain of the mouse when both compounds are given in vivo. Since a variety of studies indicate that dopaminergic (DA) systems are involved in the action of phencyclidine, it seemed that DA systems might mediate the enhancement of the binding of QNB by phencyclidine. The results of the present studies demonstrate that the neurotoxin 6-hydroxydopamine, the DA D1 antagonist (R)-8-[chloro]-2,3,4,5-tetrahydro-3-methyl-5-phenyl-1-H-3-benzazepine-7- ol) (SCH23390), the DA D1 agonist p-dibenzylamino benzonitrile (SKF38393) and the DA D2 antagonist spiperone, all failed to modify the ability of phencyclidine to enhance the accumulation of QNB in brain. Quinpirole, a DA D2 agonist diminished accumulation of QNB after phencyclidine, however, this effect was confounded by quinpirole decreasing the non-specific binding of QNB, as well as increasing concentrations of QNB in plasma, when given with phencyclidine. In contrast, haloperidol, a DA antagonist, decreased the enhanced specific binding of QNB after phencyclidine without any apparent confounding influences. Since haloperidol is also known to strongly inhibit the binding of ligands to the sigma receptors, it was hypothesized that the actions of phencyclidine and haloperidol on the binding of QNB in vivo may be through sigma receptors rather than DA systems. PMID- 2569177 TI - Paralyzed with pain: the need for education. AB - This report surveyed the pharmacologic knowledge of the physician housestaff and intensive care nurses regarding the analgesic and anxiolytic effects of narcotics, benzodiazepines and neuromuscular blockers. The results demonstrated a commonly held misconception that muscular paralysis is a calm and painless state. The authors instituted an educational program stressing the need for analgesic and anxiolytic medications in conjunction with paralytic agents. PMID- 2569178 TI - Relationship between cardiovascular neurones and descending antinociceptive pathways in the rostral ventrolateral medulla of the cat. AB - It has been previously reported that injection of neuroexcitatory compounds into the rostral ventrolateral medulla (RVLM) can produce an inhibition of nociceptive reflexes, often associated with a rise in arterial blood pressure. The aim of this study was to determine whether the subretrofacial (SRF) nucleus, which is a highly circumscribed group of cells within the RVLM known to play a major role in cardiovascular regulation also has an antinociceptive function. In barbiturate anaesthetised and paralysed cats, unilateral microinjections of the neuroexcitatory compound sodium glutamate (8-20 nl of 0.5 M solution) into the SRF nucleus produced large increases in mean arterial pressure but had only small and inconsistent effects on the simultaneously measured ventral root responses to stimulation of primary afferent C-fibres. On the other hand, glutamate microinjections into RVLM sites closely adjacent to the SRF nucleus, or into the nucleus raphe magnus, produced powerful inhibition of the C-fibre evoked response in the ventral root which was accompanied by no or only small changes in arterial pressure. It is concluded that the SRF pressor cells do not exert any control over nociceptive spinal reflexes, but that such a function may be served by cells in closely adjacent parts of the RVLM. Moreover, the method of recording C-fibre evoked responses in ventral roots as a measure of the magnitude of nociceptive spinal reflexes, combined with the glutamate microinjection procedure, was shown to have a sufficient resolution to allow an accurate mapping of the location of antinociceptive cell groups within the ventrolateral medulla. PMID- 2569179 TI - Management of burn pain in children. AB - In spite of the many possible methods of pain control in the burned child satisfactory pain management may still be a problem, at times formidable. The most fruitful approach would seem to be frequent assessment of pain in the individual patient with a readiness to try alternative or additional measures when relief seems inadequate. In this way the most effective analgesic agent(s), route(s), and frequency of administration, as well as nonpharmacologic methods, can be determined for each child. Among those able to speak, pain estimation is usually easily accomplished. In infants and those intubated for supported ventilation, however, the task is more difficult. Nevertheless, careful observation of physiologic signs such as heart rate and blood pressure, facial expressions, body movement and position, and the quality of an infant's cries may in sum be sufficient to evaluate the intensity of pain. Monitoring of analgesic plasma levels to ascertain that they are within the ranges established for good analgesia and even determination of beta-endorphin blood levels may also aid in judging the adequacy of analgesia. By tailoring pain management methods to the needs of each child it may be possible to keep pain at acceptable levels in victims of burn injury. PMID- 2569180 TI - The neuroanatomy, neurophysiology, and neurochemistry of pain, stress, and analgesia in newborns and children. AB - Beginning with a brief description of mature anatomic pathways and neurotransmitters in the "pain system," this article details their development in the human fetus, neonate, and child. Special emphasis is given to the basic mechanisms and physiologic effects of opioid analgesia. The clinical implications of these data are described, particularly with regard to the maintenance of cardiovascular stability and hormonal-metabolic homeostasis in newborns and children undergoing surgery or other forms of stress. PMID- 2569182 TI - Hypnotherapy: a cyberphysiologic strategy in pain management. AB - Children have the capacity to learn cyberphysiologic strategies in pain management. Training in such strategies can be accomplished in one to four training sessions, depending on the type of pain problems. Even in an acute emergency situation, children can benefit from informal hypnotic and biofeedback techniques. Both clinical and laboratory studies confirm the ability of children to self-regulate specific autonomic functions such as control of peripheral temperature and GSR. Such skills can be used to demonstrate to children the connections between changes in thinking and changes in body responses and pave the way for development of skills in self-regulation of pain. PMID- 2569181 TI - Pharmacologic management of pain in children and adolescents. AB - In the management of chronic pain conditions, the combination of pharmacologic measures with physical and psychologic modalities becomes even more important. A pain clinic and pain consultation service are one model that facilitates this combined approach. Initial management of mild to moderate pain begins with nonopioid analgesics such as acetaminophen and NSAIDs. Persistent severe pain of a neuropathic character merits careful trials of antidepressants or anticonvulsants. Traditionally, use of opioids for chronic pain not due to cancer has been discouraged for adults as well as children. Recently, this view was challenged by reports by Portenoy and Foley and by Taub, who followed a group of adults with chronic pain due to a variety of conditions. They found that the majority of these patients, if managed with opioids on a regular schedule as part of an overall treatment program, could be made comfortable and were able to increase their level of functioning for several years. In general, dosage escalation and compulsive drug-seeking behaviors were not seen. Since this report was retrospective and did not involve children, caution must be applied in extrapolating these findings to children. For example, remarkably little is known about the effects of chronic opioid administration in childhood on growth and development. Certainly, this issue deserves further study before general recommendations can be made. It seems prudent to emphasize the importance of maximizing nonpharmacologic and nonopioid approaches in the management of chronic pain in children prior to embarking on long-term use of opioids. PMID- 2569183 TI - Pediatric postoperative pain management. AB - This article reviews methods to relieve postoperative pain in most children. It also discusses the major barriers to treatment and considers the provision of opioids via a painless route as an alternative to the more usual intramuscular (and painful) route. PMID- 2569184 TI - The management of pain associated with pediatric procedures. AB - This article provides guidelines for the psychological and pharmacologic management of pain and anxiety for children undergoing medical procedures. The goals of intervention are presented, as well as issues warranting consideration in planning intervention to reduce procedure-related distress. PMID- 2569185 TI - The treatment of pain in the emergency department. AB - Our approach to the management of fear and pain in the pediatric emergency department is presented. Tricks to attempt the gaining of rapport with frightened children in pain are noted, with emphasis on a developmental approach. The use of analgesic medications, local anesthetics, ketamine, and nitrous oxide as appropriate to emergency situations is outlined. Lastly, the guidelines of the American Academy of Pediatrics for outpatient sedation are reviewed. PMID- 2569186 TI - The treatment of cancer pain in children. AB - In summary, expertise is currently available to successfully manage most pediatric cancer pain; it behooves the responsible physician to be sufficiently familiar with the available methodology and to appropriately deliver state-of-the art management to the child to ensure optimal care. PMID- 2569188 TI - 'A monumental challenge'. PMID- 2569187 TI - Effects of (-)baclofen on inhibitory neurons in the guinea pig hippocampal slice. AB - Intracellular recordings were made from electrophysiologically identified inhibitory neurons in the dentate hilus. (-)Baclofen (0.1-0.5 mumol/l), applied by the bath, strongly hyperpolarized inhibitory neurons, reduced their input resistance and induced outward currents under voltage clamp at holding potentials of -60 mV in cells recorded with KCl-filled electrodes. Increasing the ( )baclofen concentration (up to 1 mumol/l) did not increase the amplitude of the outward current, but increased its duration. (-)Baclofen depressed Cl-dependent IPSPs evoked by perforant path stimulation in inhibitory neurons, granule cells and CA3 neurons. In the case of inhibitory neurons and CA3 neurons, depression of IPSPs, membrane hyperpolarization and increase in membrane conductance concurred. All effects were blocked by BaCl2 (1 mmol/l) in the superfusate. In the case of granule cells, depression of IPSPs by (-)baclofen outlasted an only small membrane hyperpolarization, conductance increase or outward current. High concentrations (up to 10 mumol/l) of (-)baclofen depressed evoked IPSPs of granule cells for an extended period of time, but the other effects remained small and transient. IPSPs elicited in granule cells by microdrop application of glutamate to the dentate hilus were also blocked by (-)baclofen, but spontaneous IPSPs were only reduced in amplitude. We suggest that the blockade of GABAA receptor-mediated IPSPs of hippocampal neurons by the GABAB receptor agonist ( )baclofen can be explained by a K-dependent hyperpolarization of inhibitory neurons. PMID- 2569189 TI - On the topically effective ocular hypotensive properties of ICI 147,798, a natriuretic beta-adrenoceptor antagonist, in rabbits. AB - The ocular antihypertensive effects of ICI 147,798, a natriuretic, non-selective beta-blocker lacking local anaesthetic and intrinsic sympathomimetic activities, were evaluated in unanaesthetized rabbits with water-load-induced ocular hypertension. The racemate was approximately 20 times more potent as a beta blocker than its d-isomer. The magnitude of reductions in intraocular pressure (IOP) elicited by both compounds was equal following topical administration of an equal dose-concentration (100-300 microliters, 0.5% or 0.75%) to one or both eyes, and the activities were similar to those obtained with timolol. A five-hour duration of the IOP lowering effect was achieved by ICI 147,798 and timolol in this rabbit model. ICI d-147,798 exhibited a shorter duration, which could be prolonged by increasing the concentration of the ophthalmic solution. Timolol caused a pronounced reduction of isoproterenol-induced tachycardia after a single dose (200 microliters, 0.5%) or by repeating the dose twice a day for four consecutive days. Following the same dose regimen, neither ICI 147,798 nor its d isomer significantly altered the heart rate response produced by isoproterenol. These findings indicate that ICI 147,798, while effective as a potential antiglaucoma agent, does not display the characteristics of beta-blockers on heart rate effects noted with timolol. The results also cast some doubt on the contention that the IOP lowering effect of certain beta-blockers is predominantly determined by specific blockade of beta-adrenoceptors. PMID- 2569190 TI - High affinity binding of idazoxan to a non-catecholaminergic binding site in the central nervous system: description of a putative idazoxan-receptor. PMID- 2569191 TI - A methodological comparison of two formulations of temazepam in pharmacokinetic and pharmacodynamic aspects. AB - In a double-blind and cross-over study 12 healthy subjects took temazepam 20 mg in two different formulations (soft gelatine capsule or uncoated tablet) and matched placebo at one-week intervals. Plasma temazepam concentrations at 0.5, 1, 2, 3, 8, 12 and 24 hours after treatment were analyzed by gas chromatography. Psychomotor performance was measured objectively (digit symbol substitution, letter cancellation, Maddox wing test) and subjectively (visual analogue scales) before the drug intake and 1, 2 and 3 hours later and the plasma benzodiazepine concentrations were analyzed also by radioreceptor bioassay. The two different formulations were compared in pharmacokinetic and pharmacodynamic terms, and the gas chromatographic and radioreceptor assays were compared. The soft gelatine capsule produced higher peak plasma concentrations than the uncoated tablet. The computed AUCs and elimination half-lives proved to be similar after either formulation. A satisfactory correlation between the bioassayed benzodiazepine concentrations and chemically assayed temazepam was shown. In pharmacodynamic terms the results suggest a shorter and somewhat smaller subjective response for the capsule than for the tablet form. PMID- 2569192 TI - Pharmacodynamic and pharmacokinetic aspects on the transport of bronchodilator drugs through the tracheal epithelium of the guinea-pig. AB - An isolated vagus nerve-trachea tube preparation from guinea-pig was used to study the effect kinetics of bronchodilating beta-adrenoceptor agonists. The test compounds were added either into the fluid-filled lumen or into the external medium and they all inhibited, dose-dependently and completely, the vagally induced contractions. The hydrophilic compounds isoprenaline, salbutamol and terbutaline were much less potent when administered intratracheally as compared with extratracheal administration indicating a slow transport through the epithelial layer. For the lipophilic compound, D2489 (the resorcinol derivative of salmeterol), this difference was less pronounced. When terbutaline was administered as its lipophilic diisobutyrate ester prodrug, ibuterol, the difference between the routes of administration was largely eliminated. The inhibitory effect of terbutaline, but not D2489, was readily reversed by washing. Measurements of terbutaline and D2489 in the tracheal tissue and in the external medium after the intratracheal administration of the compounds support the view that a hydrophilic compound slowly passes the epithelium and is not retained in the tissue, whereas a lipophilic compound rapidly passes the epithelium and is retained by the tissue. The isolated vagus nerve-trachea tube preparation of the guinea-pig is well suited for the concommitant study of pharmacodynamic and pharmacokinetic properties of bronchodilator drugs. PMID- 2569193 TI - Blood gas tensions, acid-base status and cardiovascular function in miniature swine anaesthetized with halothane and methoxyflurane or intravenous metomidate hydrochloride. AB - Gottingen mini-pigs were given atropine and anaesthetized with one of the following drug combinations: 1) azaperone + metomidate, 2) acepromazine + thiopentone for endotracheal intubation, followed by halothane + methoxyflurane in oxygen. Respiration rate, arterial O2 and CO2 tensions, arterial pH, mean arterial blood pressure and heart rate were recorded at 15 min. intervals from 30 to 120 min. after the induction of anaesthesia. The azaperone-metomidate combination, which requires continuous intravenous access, caused a mild respiratory acidosis, but acid-base status was stable, whereas blood pressure and heart rate rose steadily throughout the observation period. The second combination depressed respiratory and cardiovascular function, but the condition of the animals did not deteriorate further during 2 hrs of surgical anaesthesia. Endotracheal intubation for inhalation anaesthesia should, however, be performed after a single injection of metomidate, as thiopentone tended to cause respiratory arrest. PMID- 2569194 TI - Cardiovascular function measured by ultrasound Doppler in healthy young men after ingestion of dextropropoxyphene napsylat. AB - In a double blind cross-over study 10 healthy male volunteers were given either 300 mg dextropropoxyphene napsylat (DP) or placebo daily for 16 days. The serum levels of DP and the metabolite nordextropropoxyphene were measured on day 3, 6 and 16. Haemodynamic measurements were made on day 1 and day 16, both at rest and during exercise. The measurements were made non-invasively, with a pulsed ultrasound Doppler. Blood pressure, heart rate, velocity, cardiac output, left cardiac work, increased during work, but showing no significant differences between the groups. The systolic time intervals were also measured by the ultrasound Doppler. The preejection period increased significantly in the DP group, whereas the ratio preejection period/left ventricular ejection time which reflects the contractility of the heart did not differ significantly. It is concluded that DP taken daily in a normal dose for 16 days did not affect the heart function in healthy young men. PMID- 2569195 TI - Haemogregarina sp. (Apicomplexa: Adeleorina) from the gecko Tarentola annularis in the Sudan: fine structure and life-cycle trials. AB - The ubiquitous gecko Tarentola annularis in the area around Khartoum, Sudan, was found to be infected with Haemogregarina sp., with merogonic stages in its pulmonary endothelial cells and gamonts in its erythrocytes. The fine structure of Haemogregarina meronts, merozoites and gamonts revealed great conformity with the micromorphology of similar stages of other apicomplexan parasites. The unsuccessful transmission of Haemogregarina gamonts to the mosquitoes Aedes aegypti, Culex quinquefasciatus and Anopheles stephensi is also reported. PMID- 2569196 TI - Assessment of pancreatic islet-cell population in the hyperglycemic athymic nude mouse: immunohistochemical, ultrastructural, and hormonal studies. AB - Previous studies in diabetic animal models have demonstrated altered pancreatic islet-cell populations. To further characterize the diabetic syndrome in our athymic nude mouse colony, we studied the population of endocrine cells in pancreatic islets of 4-week-old normoglycemic and 8-week-old hyperglycemic athymic nude (nu/nu) mice using immunohistochemistry, morphometry, and electron microscopy. In normoglycemic 4-week athymic nu/nu mice, the proportions of B (insulin-secreting) cells and A (glucagon-secreting) cells were similar to those in control Balb/c mice; however, the D (somatostatin-secreting) cells were significantly decreased in nu/nu mice. The populations of B and A cells appeared to be normal in hyperglycemic 8-week-old nu/nu mice while there was a significant increase in the proportion of D cells when compared with the proportion in Balb/c mice. Electron microscopic studies indicated that the appearance of B and A cells was similar in the 8-week-old hyperglycemic nu/nu and in controls; however, the D cells appeared to be enlarged and were finely packed with electron-dense secretory granules. Radioimmunoassays of the pancreatic content (micrograms/g fresh pancreas) of insulin, glucagon, and somatostatin in pancreata in 8-week-old normal Balb/c and hyperglycemic athymic nude mice were similar; however, the somatostatin content was significantly increased in the 8-week-old hyperglycemic nu/nu mice compared with age and sex-matched controls. These results demonstrate an altered D cell population and an increase in somatostatin levels in the pancreatic islets of the hyperglycemic athymic nude mouse animal model.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569197 TI - The Wellcome lecture, 1988. Muscular dystrophy: a time of hope. AB - Duchenne muscular dystrophy (DMD) and its less severe allele Becker muscular dystrophy (BMD) are progressive muscle-wasting disorders of children. DMD is characterized by rapid loss of muscle fibres and the ensuing weakness results in lost mobility and eventual premature death. Despite extensive research for many years, the basic underlying biochemical defect has remained elusive. Here I try to demonstrate how the powerful techniques of molecular genetics can be used to gain a further understanding of this particular disorder and how, in principle, the techniques can be applied to the other 3000 human genetic disorders that are so far uncharacterized. Once the chromosomal map position of DMD was established, the locus that was being disrupted by mutation could be identified and the encoded protein product predicted from the nucleotide sequence of the RNA transcript. This has led to the identification of a previously uncharacterized protein named dystrophin. As the normal function of dystrophin is determined, more accurate clinical diagnosis of DMD and BMD should result and potential approaches to therapy should be designed. PMID- 2569198 TI - The Florey lecture, 1988. From egg to embryo: the initiation of cell differentiation in Amphibia. AB - Some of the principles by which different cell types first arise at the beginning of animal development are illustrated by muscle cell formation in Amphibia. If the nucleus of a differentiated muscle cell is transplanted to an enucleated egg, some of the resulting embryos develop into tadpoles with a wide range of normally differentiated cells. These experiments show that genes undergo major changes in activity as a response to components of egg cytoplasm. Two fundamental mechanisms account for the regional activation of genes in early embryos. One involves the effect of localized 'determinants' in egg cytoplasm, and the other concerns cell interactions or embryonic induction. Both these mechanisms seem to be responsible for muscle cell formation in amphibian development. The old problem of embryonic induction has recently become accessible to analysis at the molecular level, especially in the case of the mesoderm or muscle-forming induction. This has been greatly facilitated by using a sensitive and quantitative assay to detect the first transcripts of muscle genes a few hours after the start of induction. The role of early events and of interactions among like cells during response to induction is discussed. In analysing specific gene activation following induction, DNA injection into fertilized eggs has shown that a very small part of the cardiac actin gene promoter is sufficient to enable it to respond to induction. Although the experimental work summarized here has been done on amphibian embryos, which are more suitable than other embryos for embryological manipulation, the conclusions reached are believed to be generally applicable to the development of other organisms. PMID- 2569199 TI - Epithelial action potentials in embryos of the Australian lungfish. AB - The epithelial cells of the skin of embryonic Australian lungfish (Neoceratodus forsteri) between stages 30 and 39, are mechanosensitive and excitable, showing overshooting action potentials 400-800 ms long with a rapid rise followed by a slower repolarization (usually with a shoulder on the repolarizing phase), which propagate at around 10 mm s-1. This skin impulse system is very similar to that found in embryonic and larval Amphibia; the significance of this similarity is discussed. PMID- 2569201 TI - The stability of ecosystems. AB - Stability criteria and phase boundaries for complex ecosystems are obtained and contrasted with previously studied scenarios. The stability of such systems is determined by the behaviour of the largest eigenvalue of matrices governing the response of the system to small perturbations. As a result we show that ecosystems with unstructured cooperative interactions between arbitary species can be less stable than had been previously determined. We also examine hierarchical ecologies, and demonstrate their increased stability under certain conditions. PMID- 2569200 TI - Responses of synchronous L5178Y S/S cells to heavy ions and their significance for radiobiological theory. AB - Synchronous suspensions of the radiosensitive S/S variant of the L5178Y murine leukaemic lymphoblast at different positions in the cell cycle were exposed aerobically to segments of heavy-ion beams (20Ne, 28Si, 40Ar, 56Fe and 93Nb) in the Bragg plateau regions of energy deposition. The incident energies of the ion beams were in the range of 460 +/- 95 MeV u-1, and the calculated values of linear energy transfer (LET infinity) for the primary nuclei in the irradiated samples were 33 +/- 3, 60 +/- 3, 95 +/- 5, 213 +/- 21 and 478 +/- 36 keV microns 1, respectively; 280 kVp X-rays were used as the baseline radiation. Generally, the maxima or inflections in relations between relative biological effectiveness (RBE) and LET infinity were dependent upon the cycle position at which the cells were irradiated. Certain of those relations were influenced by post-irradiation hypothermia. Irradiation in the cell cycle at mid-G1 to mid-G1 + 3 h, henceforth called G1 to G1 + 3 h, resulted in survival curves that were close approximations to simple exponential functions. As the LET infinity was increased, the RBE did not exceed 1.0, and by 478 keV microns-1 it had fallen to 0.39. Although similar behaviour has been reported for inactivation of proteins and certain viruses by ionizing radiations, so far the response of the S/S variant is unique for mammalian cells. The slope of the survival curve for X-photons (D0: 0.27 Gy) is reduced in G1 to G1 + 3 h by post-irradiation incubation at hypothermic temperatures and reaches a minimum (Do: 0.51 Gy) at 25 degrees C. As the LET infinity was increased, however, the extent of hypothermic recovery was reduced progressively and essentially was eliminated at 478 keV microns-1. At the cycle position where the peak of radioresistance to X-photons occurs for S/S cells, G1 + 8 h, increases in LET infinity elicited only small increases in RBE (at 10% survival), until a maximum was reached around 200 keV microns-1. At 478 keV microns-1, what little remained of the variation in response through the cell cycle could be attributed to secondary radiations (delta rays) and smaller nuclei produced by fragmentation of the primary ions. PMID- 2569202 TI - The determination of kinetic parameters for carrier-mediated transport of non labelled substrate analogues: a general method applied to the study of divalent anion transport in placental membrane vesicles. AB - A method is described that allows the determination of kinetic parameters for carrier-mediated transport of unlabelled compounds. The ability of these compounds to relieve the inhibition of labelled substrate efflux produced by addition of an external competitive inhibitor is studied. The method is of general applicability and does not depend upon any intrinsic asymmetry in the ratio of the rates of translocation of the loaded and unloaded carrier. In this paper it is used in a study of the human placental sulphate transporter. Experiments on brush-border membrane vesicles show the method to predict quantitatively kinetic parameters for unlabelled sulphate entry. Further analysis shows this carrier to have a broad specificity for other oxyanions (selenate, tung-state, molybdate and chromate) with the following selectivity sequence: for rate of translocation, SO4, SeO4 greater than WO4 greater than MoO4 much greater than CrO4; for binding affinity, CrO4 greater than MoO4, WO4 greater than SO4, SeO4. PMID- 2569203 TI - Changes in chlorpromazine induced CNS effects when associated with other neuroleptics and anxiolytics. AB - A set of test (General behavior, traction, chimney, rotarod, conditioned reflexes, curiosity and potentiation of barbiturate sleep) were given in addition to studying the hypothermia of associations of chlorpromazine with various neuroleptic and one anxiolytic (Perphenazine, clothiapine, haloperidol and diazepam), which demonstrated an evident enhancement of the depressant effects of chlorpromazine when combined with clothiapine and haloperidol. PMID- 2569204 TI - The antigenic structure of poliovirus. AB - We have solved the structure of the Mahoney strain of type 1 and the Sabin (attenuated vaccine) strain of type 3 poliovirus by X-ray crystallographic methods. By providing a three-dimensional framework for the interpretation of a wealth of experimental data, the structures have yielded insight into the architecture and assembly of the virus particle, have provided information regarding the entry of virus into susceptible cells, and defined the sites on the virus particle that are recognized by neutralizing monoclonal antibodies. Thus locating mutations in variants selected for resistance to neutralizing monoclonal antibodies has defined three antigenic sites of the surface of the virion, and provided clues as to the mechanisms by which viruses escape neutralization. Finally, comparison of the structures of the two strains, together with analysis of sequences of many poliovirus strains, have begun to define the structural changes associated with serotypic differences between polioviruses. PMID- 2569205 TI - Immune recognition of influenza virus haemagglutinin. AB - Haemagglutinin glycoproteins are the components of influenza virus membranes against which infectivity-neutralizing antibodies are directed. Sequence analysis of natural and laboratory-selected variant haemagglutinins indicates the regions of the molecule recognized by antibodies and by helper T cells; the identity of these regions and the relations between them are discussed. PMID- 2569206 TI - Studies of structure and specificity of some antigen-antibody complexes. AB - By using X-ray diffraction and immunochemical techniques, we have exploited the use of monoclonal antibodies raised against hen egg lysozyme (HEL) to study systematically those factors responsible for the high specificity of antigen antibody interactions. HEL was chosen for our investigations because its three dimensional structure and immunochemistry have been well characterized and because naturally occurring sequence variants from different avian species are readily available to test the fine specificity of the antibodies. The X-ray crystal structure of a complex formed between HEL and the Fab D1.3 shows a large complementary surface with close interatomic contacts between antigen and antibody. Thus single amino acid sequence changes in heterologous antigens give antigen-antibody association constants that are several orders of magnitude smaller than that of the homologous antigen. For example, a substitution of His for Glu at position 121 in the antigen is sufficient to diminish significantly the binding between D1.3 and the variant lysozyme. The conformation of HEL when complexed to D1.3 shows no significant difference from that seen in the free molecule, and immunobinding studies with other anti-HEL antibodies suggest that this observation may be generally true for the system of monoclonal antibodies that we have studied. PMID- 2569208 TI - Three-dimensional structures of influenza virus neuraminidase-antibody complexes. AB - X-ray diffraction analysis of crystals of a monoclonal Fab fragment NC41 bound to a viral antigen, influenza virus neuraminidase, shows an epitope involving five surface loops of the antigen. In addition it reveals an unusual pairing pattern between the domains of light and heavy chains in the variable module of the antibody. We interpret this result to imply that association with antigen can induce changes in the quaternary structure of the Fab, through a sliding of domains at the variable light/variable heavy chains (VL-VH) interface. In addition, Fab binding has altered the conformation of some of the surface loops of the antigen. The structure of the NC10 Fab-neuraminidase complex has now also been solved. It binds an epitope that overlaps the NC41 epitope. In this structure, there is no electron density for the C-module of the Fab fragment, implying it is disordered in the crystal lattice. The implications of these, and other antibody-antigen structures, for immune recognition are discussed. PMID- 2569207 TI - Synthetic site-directed ligands. AB - Complexes of nucleotides, peptides and aromatic hapten-like compounds with immunoglobulin fragments were studied by X-ray analysis. After tri- or hexanucleotides of deoxythymidylate were diffused into triclinic crystals of a Fab (BV04-01) with specificity for single-stranded DNA, extensive changes were detected throughout the structure of the protein. The Fab co-crystallized with a tri- or pentanucleotide in a different space group (monoclinic), an observation sometimes correlated with alterations in the structure of the 'native' protein. Structural analyses of the co-crystals are in progress for direct comparisons with the unliganded Fab. In crystals of a human (Mcg) Bence-Jones dimer, synthetic opioid peptides, chemotactic peptides or dinitrophenyl (DNP) derivatives could be diffused into a large conical binding cavity. The conformations of both the ligand and the protein were usually altered during the binding process. At the base of the cavity tyrosine residues could be displaced like trap-doors to permit entry of some opioid peptides and DNP compounds into a deep binding pocket. In co-crystals of the dimer and bis(DNP)lysine, two ligand molecules were bound in tandem, one in the main cavity and the second in the deep pocket. One ligand adopted an extended conformation, with the epsilon-DNP ring near the floor of the main cavity and the alpha-DNP group in solvent outside the binding site. There were no significant conformational changes in the protein. In contrast, the second ligand was very compact, with DNP rings immersed in the deep pocket, and the binding site was expanded to accommodate the oversized ligand. Peptides designed to be specific for the main cavity were incrementally constructed from minimal binding units by M. Geysen, G. Trippick, S. Rodda and their colleagues. A pentapeptide optimized for binding by this method was diffused into a crystal of the dimer and found by Fourier difference analysis to lodge exclusively in the main cavity as predicted. Binding regions in the BV04-01 Fab and the Mcg dimer were markedly different in size and shape. The Fab had a groove-type site, in which a layer of sidechains acted like a false floor over regions analogous to the cavity and deep pocket of the Bence-Jones dimer. PMID- 2569209 TI - A possible basis for major histocompatibility complex-restricted T-cell recognition. AB - Four distinct T-cell antigen-receptor gene loci have now been identified and partly characterized: alpha, beta, gamma and delta. All of these loci can rearrange in an immunoglobulin-like fashion and express polypeptides that contribute to either alpha:beta or gamma:delta T-cell receptor-CD3 complexes. Surprisingly, the T-cell receptor (TCR) delta coding regions are located entirely, or almost entirely, within the TCR alpha locus and share at least some of the V region gene segments, thus at least partly linking the two different types of receptor heterodimers. Analysis of potential T-cell receptor diversity, particularly that of the delta chain, indicates a striking concentration of somatic polymorphism in the V-J junctional region of the two heterodimers, four to six orders of magnitude higher than similar calculations for immunoglobulin light- and heavy-chain combinations. In contrast, the number of possible V region combinations in T-cell receptors is one hundredth to one thousandth that of immunoglobulins. TCR alpha: beta heterodimers are known to recognize many possible fragments of antigens embedded in the peptide-binding clefts of a relatively small number of major histocompatibility complex (MHC) molecules. Thus it is attractive to speculate that the V-J junctional portions of both types of T cell receptor contact peptide antigens, whereas the remaining diversity regions contact the MHC. This contention is supported by molecular modelling studies and has interesting implications for the evolution of antigen-receptor genes. PMID- 2569210 TI - Structural requirements and biological significance of interactions between peptides and the major histocompatibility complex. AB - Previous studies indicate that T cells recognize a complex between the major histocompatibility complex (MHC) restriction-element and peptide-antigen fragments. Two aspects of this complex formation are considered in this paper: (1) what is the nature of the specificity of the interactions that allows a few MHC molecules to serve as restriction elements for a large universe of antigens; and (2) what is the relative contribution of determinant selection (i.e. antigen MHC complex formation) and T-cell repertoire in determining the capacity of an individual to respond to an antigen? By analysing single amino acid substitution analogues of a peptide antigen (Ova 325-335) as well as by analysing the structural similarities between unrelated peptides capable of binding to the same MHC molecule, we have been able to document the very permissive nature of the antigen-MHC interaction. Despite this permissiveness of binding, it is possible to define certain structural features of peptides that are associated with the capacity to bind to a particular MHC specificity. With respect to the question of the relative role of 'determinant selection' and 'holes in the T-cell repertoire' in determining immune responsiveness, we present data that suggest both mechanisms operate in concert with one another. Thus only about 30% of a collection of peptides that in sum represent the sequence of a protein molecule were found to bind to Ia. Although immunogenicity was restricted to those peptides that were capable of binding to Ia (i.e. determinant selection was operative), we found that about 40% of Ia-binding peptides were not immunogenic (i.e. there were also 'holes in the T-cell repertoire'). PMID- 2569211 TI - The conformational restriction of synthetic peptides, including a malaria peptide, for use as immunogens. AB - A new strategy is advanced for the conformational restriction of peptidyl immunogens. Our approach is to replace putative amide-amide hydrogen bonds with covalent hydrogen-bond mimics. Because on average every other amino acid in a protein engages in this bond, the syntheses of diversely shaped peptides can be contemplated. Synthetic methods for introducing a potential hydrogen-bond mimic into a peptide with alpha-helical potential is reported and the structural consequences are discussed. The replacement of the hydrogen bond with a chemical link will modify as well as shape the peptide. To explore the consequences of these changes, a potential synthetic vaccine for malaria, the repeating tetrapeptide Asn-Pro-Asn-Ala, was conformationally restricted. Antibodies to the shaped malarial peptide showed a strong cross reaction with Plasmodium falciparum sporozoites. PMID- 2569212 TI - Pathways of cardiac sensitivity. PMID- 2569213 TI - Benzodiazepines and vigilance performance: a review. AB - The literature on the effects of anxiolytic and hypnotic drugs on performance in tasks requiring sustained attention is confusing. This review is an attempt to evaluate the usefulness of vigilance tasks in the assessment of adverse effects of benzodiazepines. The long, monotonous, character of these tasks may be more relevant to many tasks performed in everyday life than the short, and often stimulating, tasks commonly employed in test batteries. From 37 available studies, 26 were examined in detail. In young, normal volunteers, vigilance tasks were found to be sensitive, often dose dependently, to the impairing effects of drugs, even in low doses (2.5 mg diazepam). With these subjects the tasks may be successfully used to compare different benzodiazepines with respect to residual activity. Both accuracy and speed of performance appear to be affected. However, in people actually using the drugs ("patients"), adverse effects on performance are usually not found. There is no evidence that benzodiazepines aggravate the vigilance decrement occurring under normal conditions. They do affect overall level of perceptual sensitivity, but show less effects on response criterion. The drugs do not seem to interact with anxiety or sleep quality in their effect on performance, but there are few studies with patients, and the assessment of anxiety is not without problems. It is unlikely that impairments in vigilance are simply a byproduct of global, sedative effects, but there is uncertainty regarding measures of general sedation. Developing tolerance with repeated doses has been noted only occasionally, but the opposite of tolerance, aggravated impairment, has also been reported. PMID- 2569214 TI - Controlled comparison of two doses of milnacipran (F 2207) and amitriptyline in major depressive inpatients. AB - A multicenter study compared the antidepressant efficacy and the tolerance of two doses of milnacipran (50 mg and 100 mg/day) and amitriptyline (150 mg/day) in three parallel groups of 45 major depressive inpatients defined by Research Diagnostic Criteria. After a wash-out period of 4-7 days on placebo with lorazepam and/or nitrazepam if necessary, patients were randomly assigned to a daily dose of milnacipran 50 mg, milnacipran 100 mg or amitriptyline 150 mg reached on the 5th day and then stable over a 4-week period, with weekly assessments by means of the Montgomery and Asberg depression scale, the Hamilton depression scale, the Clinical Global Impressions (CGI) and the Target Emergent Signs and Symptoms. Results showed significant superiority of both milnacipran 100 mg/day and amitriptyline over milnacipran 50 mg/day at the end of the treatment period. However, amitriptyline induced a nonsignificant trend toward more rapid improvement after 2 weeks of treatment, mainly based on items related to insomnia, supporting more sedative properties of amitriptyline as compared to milnacipran. Anticholinergic side-effects were significantly lower with milnacipran than with amitriptyline, explaining why milnacipran 100 mg exhibited at the end of the treatment period, a nonsignificantly better efficacy index on the CGI. Moreover, in contrast to milnacipran, amitriptyline was responsible for a significant decrease in blood pressure and a significant weight gain. PMID- 2569215 TI - Behavioral effects of (+-) 3,4-methylenedioxyamphetamine (MDA) and (+-) 3,4 methylenedioxymethamphetamine (MDMA) in the pigeon: interactions with noradrenergic and serotonergic systems. AB - The effects of three amphetamine analogs were assessed in pigeons key pecking under a multiple 3-min fixed-interval (FI), 30 response fixed-ratio (FR) schedule of food presentation. At doses between 0.1 and 10.0 mg/kg, (+-) 3,4 methylenedioxyamphetamine (MDA), (+-) 3,4-methylenedioxymethamphetamine (MDMA), and (+-)-N-ethyl-3,4-methylenedioxyamphetamine (MDE) either had no effect or decreased response rates in both components of the multiple schedule in a dose dependent manner. MDA was at least 1 log unit more potent than the other two compounds. Metergoline (0.1-1.0 mg/kg), a serotonin (5-HT) antagonist with comparable affinity for the 5-HT1 and 5-HT2 receptor subtypes, blocked the rate decreasing effects of 3.0 mg/kg MDA in both components of the multiple schedule, but did not affect the MDMA dose-response curve. The 5-HT2 receptor antagonist ketanserin (0.1-3.0 mg/kg) also restored FI and FR responding that was decreased by 3.0 mg/kg MDA, but had no effect on responding suppressed by MDMA. The noradrenergic alpha-1 antagonist prazosin (0.3-3.0 mg/kg) blocked the behavioral effects of 3.0 mg/kg MDMA at doses that did not attenuate MDA's rate-decreasing effects. These results indicate that although MDA and MDMA are structurally similar and have similar behavioral effects, their actions appear to be mediated through different neurotransmitter systems. PMID- 2569216 TI - Haloperidol- and SCH23390-induced dopaminergic supersensitivities are not additive in the rat. AB - The effect of chronic D-1 and/or D-2 dopamine receptor blockade on apomorphine induced behaviors was studied in rats treated for 21 days with the selective D-1 antagonist SCH23390, the predominantly D-2 antagonist haloperidol, and the combination of the two drugs at the same daily doses (0.1 and 1 mg/kg respectively). Apomorphine (0.3 mg/kg) 4 days following the last injection of the drugs increased (49-70%) stereotypic behavior in all animals as compared to saline-treated controls. Although the SCH23390-induced increase was lower than haloperidol-induced supersensitivity, stereotypies after combined administration of both drugs did not differ significantly from either, suggesting that the effects of the two drugs are not additive. Underlying receptor changes and modified D-1/D-2 receptor interactions may account for the participation of both receptor subtypes to the development of neuroleptic-induced dopaminergic supersensitivity. PMID- 2569218 TI - Further evidence for two directions of D-1:D-2 dopamine receptor interaction revealed concurrently in distinct elements of typical and atypical behaviour: studies with the new enantioselective D-2 agonist LY 163502. AB - The new, extremely potent and enantioselective D-2 agonist LY 163502 failed to induce compulsive stereotyped behaviour. Very low doses (3-6 micrograms/kg) inhibited spontaneous sniffing and locomotion, while higher doses (12-50 micrograms/kg) induced episodes of non-stereotyped sniffing and chewing; these actions showed complete enantioselectivity. Up to 200-fold higher doses modestly induced only locomotion. Responsivity to LY 163502 was enantioselectively blocked by the selective D-2 antagonist R-piquindone. This responsivity was also enantioselectively blocked by the selective D-1 antagonist R-SK&F 83566 but, additionally, episodes of atypical limb/body jerking behaviour were released; thus, LY 163502 induced such jerking only when tonic D-1 activity was suppressed. These data extend our notion that there may be at least two forms of functional interaction between D-1 and D-2 receptor systems: one cooperative, as in the regulation of typical sniffing, and another oppositional, as in the regulation of atypical jerking. PMID- 2569219 TI - Increased number of cardiac adrenergic receptors following local heart irradiation. AB - The effect of local X irradiation on cardiac alpha and beta receptors was studied in Wistar rats. Animals were given local heart irradiation with single doses of 15 or 20 Gy and were examined after a range of latency times of 7 to 400 days. Using the radioactive ligands [3H]CGP-12177 and [3H]prazosin, the maximal binding capacity was determined from saturation experiments. At 7 days after 20 Gy the maximal binding capacity of both alpha and beta receptors was reduced to below the level of untreated control animals. Subsequently it rose continually to a maximum of 160% of the control level for beta receptors and 130% for alpha receptors at 400 days postirradiation. The antagonist affinity as judged from the dissociation constant for [3H]CGP 12177 and [3H]prazosin did not change significantly. A similar effect was observed after 15 Gy. An increase in adrenergic receptors may represent an important pathogenetic link between early morphological and late functional changes in the pathogenesis of radiation induced heart disease. PMID- 2569220 TI - Expression of the c-erbB-2 proto-oncogene protein in human breast cancer. AB - The discovery of a dominantly transforming oncogene in ENU-induced rat CNS tumours has revealed in rats and man a gene highly related to the EGF receptor. The gene may be activated experimentally by mutation or overexpression such that its expression can convert immortalised but non-tumourigenic rat fibroblasts to a tumourigenic state. The mechanism by which this activation occurs is not known, nor is it known whether the molecule is itself a growth factor receptor. It is hypothesised, however, that in the light of its similarity to the EGF receptor it may be involved in growth regulation and that activation may occur by producing an aberrant growth control signal. The gene has been found to be amplified frequently and the protein it encodes overexpressed in a variety of human adenocarcinomas. Investigations are underway to attempt to reveal its role in these tumours and to assess its possible value as an indicator of tumour behaviour. Overexpression of the protein provides a target for tumour imaging and therapy. PMID- 2569221 TI - Characteristics of carrier-specific regulatory T cells. PMID- 2569217 TI - Evidence that the aversive effects of opioid antagonists and kappa-agonists are centrally mediated. AB - The role of central versus peripheral opioid receptors in mediating the aversive effects of opioids was examined by use of an unbiased place preference conditioning procedure in rats. The non-selective opioid antagonist naloxone (NLX) produced conditioned aversions for the drug-associated place after subcutaneous (SC) as well as intracerebroventricular (ICV) administration. Place aversions were also observed in response to the ICV administration of the selective mu-antagonist CTOP. In contrast, the selective delta-antagonist ICI 174,864 and the selective kappa-antagonist norbinaltorphimine (nor-BNI) (ICV) were without effect. Place aversions were also produced by central applications of the selective kappa-agonist U50,488H and the dynorphin derivative E-2078. For those opioid ligands tested, the doses required to produce place aversions were substantially lower following ICV as compared to SC administration. These data confirm that kappa-agonists and opioid antagonists produce aversive states in the drug-naive animal and demonstrate that this effect is centrally mediated. Furthermore, the ability of NLX and CTOP, in contrast to both ICI 174,864 and nor BNI, to produce place aversions suggests that the aversive effects of opioid antagonists result from the blockade of mu-receptors. PMID- 2569222 TI - T-cell-induced chronic immunoglobulin allotypic suppression in mouse. PMID- 2569223 TI - Suppression in an adoptive hapten-carrier system. PMID- 2569224 TI - Suppressor cells for cell-mediated immunity in infectious diseases. PMID- 2569225 TI - Diagnosis and treatment of normal and pathologic memory impairment in later life. PMID- 2569226 TI - [Type 2b multiple endocrine adenomatosis]. AB - The case report presented here demonstrates the typical diagnostic and therapeutic problems in a young patient with a multiple endocrine adenomatosis type 2b. The patient exhibited the complete syndrome with its endocrine and non endocrine organ manifestations (medullary thyroid cancer, bilateral pheochromocytoma, mucosal neuromas, marfanoid habitus, pes equinus, diverticulosis). First presenting symptom was a diverticulitis that led to an emergency surgical intervention because of perforation of the sigmoid colon. Perioperatively, blood pressure increases were noted that were then followed by further diagnostic measures leading to adrenalectomy, thyroidectomy and surgery for local metastases of the thyroid cancer. Family screening demonstrates so far a possible sporadic form of the disease. PMID- 2569227 TI - [The diagnosis of alcoholism]. PMID- 2569228 TI - [Agranulocytosis in rheumatoid polyarthritis treated with sulfasalazine]. PMID- 2569229 TI - T-cell response to anti-CD2 monoclonal antibodies in Down's syndrome. AB - Peripheral blood mononuclear cells from 10 subjects with cytogenetically documented Down's syndrome (DS) and from 10 age- and sex-matched healthy controls were assayed for their ability to proliferate in response to phytohaemagglutinin, anti-CD3 (OKT3), or anti-CD2 (T11(2) plus T11(3] monoclonal antibodies. Interleukin 2 (IL-2) receptor expression and IL-2 production in mitogen-pulsed lymphocyte cultures was also investigated in parallel. DS cells responded poorly to all the blastogenic stimuli used in this study. Under certain experimental conditions (anti-CD3 or anti-CD2 antibody stimulation), the patients' lymphocytes expressed low levels of IL-2 surface receptors and failed to produce normal amounts of this lymphokine. Studies are currently in progress in our laboratories to determine whether these defects are due to an impairment of the early signalling events surrounding the complexing of CD3, CD2, or lectin receptors to their respective ligands. PMID- 2569231 TI - Brucellosis in an inuit child, probably related to caribou meat consumption. AB - A 9-year-old Inuit boy with brucellosis is presented. The most likely source of his infection was contaminated caribou meat. Brucella suis is enzootic in Canadian caribou herds and this case indicates that natives of the Arctic are a risk group for acquiring brucellosis through the ingestion of raw caribou meat. PMID- 2569230 TI - Increased expression of leucocyte adherence-related glycoproteins by polymorphonuclear leucocytes during phagocytosis of staphylococci on an endothelial surface. AB - Phagocytosis of Staphylococcus aureus by human polymorphonuclear leucocytes (PMN) on the surface of endothelial cells is accompanied by adherence of the PMN to the endothelial surface and detachment of the endothelial cells from the culture monolayer. We studied the role of the leucocyte adherence-related glycoproteins (Leu-CAM: Mo1/LFA-1/150,95 or CD11a-c-CD18 complex) in these processes. Phagocytosis of S. aureus induced increased expression of the common beta chain (CD18) of Leu-CAM as demonstrated by flow cytometric analysis of PMN treated with a monoclonal antibody (MoAb) (CLB-LFA-1/1) directed against CD18 and fluorescein isothiocyanate (FITC)-conjugated anti-MoAb. This same MoAb also inhibited the increased adherence of the PMN to the endothelial cells which occurs during phagocytosis. Blocking of adherence during phagocytosis with MoAb CLT-LFA-1/1 had no effect on the detaching activity of the PMN on the endothelial cells. We conclude that adherence of PMN to endothelial cells during phagocytosis of S. aureus is mediated by the Leu-CAM complex. Adherence through the Leu-CAM, however, is not necessary for endothelial damage by the phagocytosing PMN. PMID- 2569232 TI - [Gastroduodenal ulcer disease: what surgical indications are left?]. AB - Between 1976, date of the introduction of anti-H2, and 1987, 716 patients were hospitalized and operated on for ulcer disease in the surgical department of this hospital. The present study shows that 1. the annual number of operations has regressed by some 30%, 2. while the number of gastric ulcers remains constant on the whole, cases of acute duodenal ulcer have diminished by half, 3. cases of chronic, recurrent, or therapy-resistent duodenal ulcer have increased some tenfold (2 out of 78 in 1976 and 14 out of 45 in 1987), 4. the same is true of perforated ulcers (12 out of 78 in 1976 and 10 out of 45 in 1987) and hemorrhages (12 out of 78 and 10 out of 45), 5. surgery for stenosis has remained constant. Proximal selective vagotomy has been the treatment of choice since 1981 in over 80% of duodenal ulcers. Operative mortality affects only elderly patients undergoing emergency surgery for complicated ulcer (two thirds perforations, one third hemorrhages). It is 2.6%. We thus confirm the reduced role of surgery in the treatment of gastric ulcer, while redefining the present surgical indications. PMID- 2569233 TI - [Campylobacter pylori colonization of the antrum: effect of gastrin, somatostatin, pancreatic polypeptide and neurotensin]. AB - Campylobacter pylori (C.p.) infection is often found in patients with antral gastritis and peptic ulcer disease. Pathophysiological links are still unclear, and we therefore tested the hypothesis whether C.p. affects the gastrointestinal peptides and thus influences gastric acid secretion and protective factors. 94 patients were examined by upper GI endoscopy and blood analyzed for gastrin, somatostatin, pancreatic polypeptide and neurotensin. Biopsies of antral mucosa were investigated for C.p. in urease testing, culture and microscopy. C.p. was found in 42 patients (45%). In microscopy all of these patients had chronic gastritis (100%). A significant increase in gastrin uninfluenced by C.p. was found in patients with antral gastritis (normal: 6.4 +/- 0.7, [n = 27]; gastritis without C.p.: 18.4 +/- 5.9 [p less than 0.02], [n = 7]; gastritis with C.p.: 10.7 +/- 2.2, [n = 22]). Somatostatin, pancreatic polypeptide and neurotensin showed no difference. PMID- 2569234 TI - Viral alteration of cell function. PMID- 2569235 TI - Genetic and pharmacological suppression of oncogenic mutations in ras genes of yeast and humans. AB - The activity of an oncoprotein and the secretion of a pheromone can be affected by an unusual protein modification. Specifically, posttranslational modification of yeast a-factor and Ras protein requires an intermediate of the cholesterol biosynthetic pathway. This modification is apparently essential for biological activity. Studies of yeast mutants blocked in sterol biosynthesis demonstrated that the membrane association and biological activation of the yeast Ras2 protein require mevalonate, a precursor of sterols and other isoprenes such as farnesyl pyrophosphate. Furthermore, drugs that inhibit mevalonate biosynthesis blocked the in vivo action of oncogenic derivatives of human Ras protein in the Xenopus oocyte assay. The same drugs and mutations also prevented the posttranslational processing and secretion of yeast a-factor, a peptide that is farnesylated. Thus, the mevalonate requirement for Ras activation may indicate that attachment of a mevalonate-derived (isoprenoid) moiety to Ras proteins is necessary for membrane association and biological function. These observations establish a connection between the cholesterol biosynthetic pathway and transformation by the ras oncogene and offer a novel pharmacological approach to investigating, and possibly controlling, ras-mediated malignant transformations. PMID- 2569236 TI - Brain region and gene specificity of neuropeptide gene expression in cultured astrocytes. AB - Astrocytes have many neuronal characteristics, such as neurotransmitter receptors, ion channels, and neurotransmitter uptake systems. Cultured astrocytes were shown to express certain neuropeptide genes, with specificity for both the gene expressed and the brain region from which the cells were prepared. Somatostatin messenger RNA and peptides were detected only in cerebellar astrocytes, whereas proenkephalin messenger RNA and enkephalin peptides were present in astrocytes of cortex, cerebellum, and striatum. Cholecystokinin was not expressed in any of the cells. These results support the hypothesis that peptides synthesized in astrocytes may play a role in the development of the central nervous system. PMID- 2569237 TI - Drugs from emasculated hormones: the principle of syntopic antagonism. AB - This lecture illustrates the early stages in the planning and discovery of propranolol, an adrenaline beta-adrenergic receptor antagonist, and cimetidine, a histamine H2-receptor antagonist--the first examples of clinically useful drugs from each of these classes. The significance of selective agonists, partial agonists, and syntopic antagonists and the importance of the bioassay and the use of molar models in the drug discovery process are discussed. For the future, an outline of potential developments in hormone-receptor concepts is offered leading to the conclusion that progress may depend on improvements in bioassays and related molar modeling. PMID- 2569238 TI - [The aged and drugs. Neuroleptic treatment and nurses' responsibility]. PMID- 2569239 TI - [Triazolam 0.25 mg versus loprazolam 1 mg in the treatment of common insomnia treated in general practice. Double-blind cross-over randomized trial]. AB - In a double-blind cross over randomized study for the first two nights efficacy and safety of triazolam 0.25 mg and loprazolam 1 mg have been compared in 67 out patients complaining of common insomnia and treated by general practitioners. After the second night patients had to choose one of the two treatments and were continuing a 3-week treatment period with the preferred one. In case of no preference they received one of the 2 drugs according to a new randomization. Cross over administration of the two drugs for the first two nights shows that with triazolam global help to get in sleep is greater (p = 0.016) and sleep latency is shorter (p = 0.07) than with loprazolam, and number of night awakenings is decreased (p = 0.02) compared to loprazolam. Patients felt more rested under triazolam (p = 0.015) than loprazolam. Triazolam (N = 31) is more frequently preferred than loprazolam (N = 19) p = 0.09. Preferred treatment continued to be effective during the following three weeks and quality of sleep improved drastically for all items compared to baseline data (p = 0.01). Both treatments are well tolerated (4 drop-outs for side-effects: 2 under each treatment). The one-week tapering period allows progressive withdrawal with rare reappearance of minimum sleep disorders (5 cases: 2 under triazolam, 3 under loprazolam). PMID- 2569240 TI - The role of the first assistant in ophthalmic surgery. AB - 1. As the number of Ambulatory Surgery Centers increases and the Medicare reimbursements for surgeon first assistants decline, the need for non-physician first assistants grows. 2. Qualifications for a first assistant include knowledge, skill, and devotion to this specialty. 3. The first assistant is part of a medical team that consists of the surgeon, the first assistant, the scrub nurse or technician, and the circulating nurse. 4. The role of the first assistant helps to increase efficiency in the operating room and enhance the quality of patient care. PMID- 2569242 TI - An update on the vertebrate homeobox. PMID- 2569241 TI - Effects of Thy-1+ cell depletion on the capacity of donor lymphoid cells to induce tolerance across an entire MHC disparity in sublethally irradiated adult hosts. AB - Thy-1+ cell depletion with anti-Thy-1.2 mAb and complement markedly reduced the capacity of C57BL/6J, H-2b bone marrow to establish mixed lymphoid chimerism and induce tolerance to C57BL/6J skin grafts across an entire MHC disparity in BALB/c, H-2d hosts conditioned with sublethal, fractionated 7.5 Gy total-body irradiation. In this model tolerance can be transferred to secondary irradiated BALB/c hosts only by cells of C57BL/6J donor, not host, genotype isolated from the spleens of tolerant hosts. Thy-1+ cell depletion abolished the capacity of C57BL/6J donor cells from tolerant BALB/c host spleens to transfer tolerance. The capacity of semiallogeneic BALB/c x C57BL/6J F1, H-2d/b donor BM and spleen cells to induce chimerism and tolerance to C57BL/6J skin grafts in BALB/c parental hosts was also reduced by Thy-1+ cell depletion. Thus the requirement for donor Thy-1+ cells cannot be explained simply on the basis of alloaggression. It is unlikely that the requisite Thy-1+ cells are nonspecific suppressor cells: Thy-1+ cell depletion had no effect on the slight but significant prolongation of third party C3H/HeJ, H-2k skin grafts in irradiated BALB/c hosts injected with allogeneic C57BL/6J or semiallogeneic BALB/c x C57BL/6J F1 BM compared to irradiated controls injected with medium only. Furthermore, injections of semiallogeneic F1 spleen cells had no significant effect on the survival of the third-party grafts, although these cells were fully capable of inducing tolerance, and their capacity to induce tolerance was significantly reduced by Thy-1+ cell depletion. The requirement for a specific population of lymphoid cells, i.e. Thy-1+, remains unexplained but suggests that donor cells might play a role in the induction or maintenance of tolerance in this model other than merely providing a circulating source of donor antigens. PMID- 2569243 TI - Autoregulation--a common property of eukaryotic transcription factors? PMID- 2569244 TI - [Cetirizine. A new non-sedative antihistaminic agent]. PMID- 2569245 TI - [Functional results after replantation/revascularization of major parts of the extremities]. AB - Twenty-five patients who were submitted to replantation or revascularization of a total of 11 upper arms, 11 forearms, one wrist and three feet were followed-up with the object of assessing the functional status. 85% of the replantated/revascularized parts survived. The follow-up examination included assessment of the level of amputation, mechanism of the injury, duration of ischaemia and the age of the patient in relation to the joint mobility achieved, strength, sensibility, cosmetic inconveniences and the return to previous employment. Out of 19 surviving upper limbs and three feet, the results were classified as excellent and good after Tamai's and Chen's methods. The best results were found after sharp lesions but the investigation revealed that even in severe avulsion lesions, it proved possible to obtain good results. It is thus not possible to establish definite directives for the indications for replantation. The final decision concerning reconstruction cannot be made until after contact with one of the departments in which replantation can be undertaken. PMID- 2569247 TI - Radionuclide renography predicts functional changes in patients with renal artery involvement by Takayasu's arteritis. AB - Renovascular hypertension is a major complication of Takayasu's arteritis, which contributes to the high mortality associated with the disease. We studied 5 patients affected by different degrees of Takayasu's arteritis to assess the usefulness of radionuclide renography in evaluating renal perfusion and function, and to predict changes induced by the disease before and after therapeutic interventions. Computer-assisted dynamic renal imaging with Tc-99m diethylenetriaminepentaacetic acid (DPTA) and I-131 orthoiodohippurate (OIH), and renal arteriography were concurrently performed in all patients. Two patients with hemodynamically insignificant renal artery stenosis showed normal perfusion and function by renography. Three patients had significant renal artery stenosis and functional changes on renography. Subsequently, two of these patients had successful therapy (one had bilateral renal artery bypass grafts, and the other had renal artery angioplasty), and both showed functional improvement at renography. Our results demonstrate that radionuclide renography is valuable in the assessment of functional changes induced by Takayasu's arteritis as well as for determining the response to therapeutic interventions. PMID- 2569246 TI - CT and MR imaging of malignant germ cell tumor of the undescended testis. AB - Preoperative localization of the impalpable undescended testis is necessary to facilitate proper surgical planning. There is an increased incidence of malignant change in the undescended testis; demonstration of malignancy before surgery will significantly alter the treatment. We describe the computed tomographic (CT) and magnetic resonance (MR) findings in 2 patients with malignant change in an intraabdominal testis. The CT scan revealed lesions with areas of low density, 1 of which had focal calcifications; MR revealed lesions of predominantly low or intermediate signal intensity on both long and short TR/TE images, with some areas of very high signal on both sequences. After initial management with chemotherapy, the residual tumor was surgically resected. In neither instance was residual normal testis demonstrated. Both CT and MR are ideal methods of examining malignant transformation of the undescended testis, because of their ability to characterize the internal structure of the organ and, in the case of MR, its capacity for multiplanar imaging. They are almost of equal value except for the ability of CT to identify calcification and of MR to diagnose hemorrhage. PMID- 2569248 TI - Identification and percentage frequency of isolated non-parenchymal liver cells (NPLC) in the mouse. AB - The aim of the present study was to identify non-parenchymal liver cells (NPLC) in B10.D2 mice and to determine their percentage frequency. The isolation of NPLC was carried out using the collagenase/pronase technique. Using functional techniques (latex phagocytosis, immunocytochemical detection of surface-bound and intracytoplasmic antigens) and morphological methods (light and electron microscopy), the following cell types were identified, and their percentage frequency in the NPLC determined: endothelial cells (50%), macrophages (23%), desmin-positive cells (14%), immunocompetent cells (10%, including T-, B-cells, pit and large vacuolated cells-both immunopositive to the asialo-GM 1 antigen) and unidentified cells (3%). These results show that, apart from the more familiar varieties of NPLC, two groups of cells exist in the liver which have not yet been fully identified and in which the immunocompetent cells predominate numerically. PMID- 2569249 TI - Endocrine cells in human Bartholin's glands. An immunohistochemical and ultrastructural analysis. AB - Endocrine cells were investigated in human Bartholin's glands by use of histochemical, immunohistochemical and ultrastructural methods. Endocrine cells represent normal constituents of these glands, being mainly distributed throughout the transitional epithelium of the major excretory duct; however, single elements are dispersed among the acinar lobules. Serotonin-, calcitonin-, katacalcin-, bombesin- and alpha-hCG-immunoreactive cells were recognized, with serotonin-immunoreactive cells predominating. Co-expression of calcitonin, katacalcin or alpha-hCG with serotonin was observed in single endocrine cells. At the ultrastructural level, these cells are richly granulated and show typical neuroendocrine features. Bartholin's glands display an endocrine profile quite similar to that of other cloacal-derived tissues. PMID- 2569250 TI - Cytotoxic effect of oxygen on the skeletal muscle of mouse diaphragm. AB - Incubation of the isolated mouse diaphragm with a high rate of oxygenation (10 ml s-1, 95% O2 + 5% CO2) causes a characteristic cellular damage with widely separated myofibrils and swollen sarcotubular system within 10 min. This damage was ameliorated by inhibitors of the hydroxyl radical (.OH), desferrioxamine, dimethyl thiourea and 120 mM mannitol, and by incubation at 8 degrees C. It was not prevented either by inhibitors of the pathway leading to sarcolemma damage (nordihydroguaiaretic acid, alpha-tocopherol, butylated hydroxytoluene) nor by agents and treatments that inhibit the oxygen paradox of cardiac muscle (glucose, omission of extracellular calcium, incubation at 30 degrees C, superoxide dismutase and catalase). Nevertheless there are similarities between these two types of damage triggered by O2 and the possibility that in both an NAD(P)H oxidase is stimulated and cytotoxic oxygen radicals are generated is discussed. PMID- 2569251 TI - Ultrastructure of Purkinje cells in the subendocardium and false tendons in early experimental myocardial infarction complicated by fibrillation in the dog. AB - The effect of regional myocardial ischemia complicated by ventricular fibrillation (VF) on the ultrastructure of subendocardial (SE) and false tendon (FT) Purkinje cells (PC) was studied in anesthetized dogs. In all cases of early ischemia with spontaneous VF, many PC exhibited ultrastructural damage as early as 2 min after the onset of ischemia. The changes noted were: intercalated disk dissociation, sarcoplasmic reticulum vacuolization (SRV), supercontraction, mitochondrial swelling, and sarcolemmal defects (rigor cells). The appearance of at least some rigor PC seemed to precede spontaneous VF, since these cells were absent from the conduction systems in control hearts in which VF was induced by electric shock or reperfusion, from hearts from sham-operated dogs, or from hearts subjected to longer periods of uncomplicated myocardial infarction. These observations indicate that alterations in SE and FTPC may play a role in the pathogenesis of sudden death due to early myocardial ischemia. The mechanism of this rapid damage of PC remains obscure. PMID- 2569252 TI - Ultrastructure of cholinergic innervation in the cirrhotic liver in guinea pigs. Neurohistochemical and ultrastructural study. AB - Hepatic cirrhosis was induced in guinea pigs by ligation of the common bile duct and innervation of the liver was studied by fluorescence histochemistry (glyoxylic acid method), acetylcholinesterase (AChE) neurohistochemistry (modified Karnovsky and Roots method), and transmission electron microscopy. In control animals the adrenergic terminals showed connections with endothelial cells, hepatocytes and fat-storing cells, but no cholinergic terminals were evident. Cirrhosis was present 6 weeks after the bile duct ligation and marked fibrosis, accompanied by bile duct proliferation, was evident in the portal areas. Numerous AChE-positive nerve fibers traversed the collagenous bundles in the fibrotic areas, and cholinergic terminals formed close contacts with fibroblasts. Each axon terminal was found to contain numerous small coreless vesicles and AChE-reaction products were confirmed in the space between a nerve terminal and a fibroblast. In contrast, fluorescence adrenergic nerve fibers and their terminals remained unchanged. This study demonstrates that parasympathetic cholinergic innervation participates in some stages in the development of hepatic cirrhosis. PMID- 2569253 TI - Adrenal cortical foci and nodules in Sprague-Dawley rats treated with N nitrosomorpholine. Light and electron microscopic findings. AB - Treatment of male Sprague-Dawley rats with N-nitrosomorpholine (NNM) in the drinking water at a dose of 120 mg/l for 7 weeks resulted in a subsequent enhanced development of focal and nodular lesions in the adrenal cortex. Sequential observation revealed that focal lesions in the zona reticularis/fasciculata or the zona glomerulosa developed both earlier and at a significantly higher incidence in animals treated with carcinogen than in untreated controls. Foci observed within or adjacent to the zona glomerulosa were all of pale cell appearance and contained large numbers of electron-dense cytoplasmic granules similar to those observed in normal granulosa cells. The foci and nodules which arose in the zona reticularis/fasciculata were, in contrast, characterized by a reduction or loss of the dense osmiophilic droplets normally seen in the cells of this region of the adrenal cortex, a pronounced increase in pleomorphic mitochondria of atypical appearance and the development of vacuoles. PMID- 2569254 TI - Unusual histochemical pattern in preneoplastic hepatic foci characterized by hyperactivity of several enzymes. AB - In a stop-experiment using the hepatocarcinogen N-nitrosomorpholine, as well as glycogenotic and related lesions, hepatocellular foci with a different histochemical pattern were identified. The outstanding features of these hepatic foci, which may progress to hepatocellular adenoma, were increased activities of mitochondrial glycerol-3-phosphate dehydrogenase (mG3PD), glycogen synthase, pyruvate kinase and glucose-6-phosphatase detected by enzyme histochemistry. Since no decrease in activity of any of the enzymes examined were seen in these foci, compared with normal liver, the term enzymatically hyperactive focus (EHF) is proposed for this type of lesion. Only at the stage of overtly nodular growth did these lesions exhibit some of the characteristic changes seen in nodules developing from glycogenotic foci, namely elevated activities of glucose-6 phosphate dehydrogenase, gamma-glutamyl transferase and glutathione-S-transferase P as well as decreased activities of adenosine-triphosphatase, glucose-6 phosphatase and adenylate cyclase. Some of these enzymes have been used widely in morphometric studies as markers for preneoplastic and neoplastic lesions. The inability to detect early EHF may lead to an underestimation of preneoplastic liver lesions in quantitative studies. Although there are apparent differences in the histochemical patterns of glycogen storing foci and early EHF, these differences tend to disappear during progression to overtly neoplastic lesions. In studies comparing the phenotypic alterations in different types of preneoplastic hepatic lesions, the recognition of EHF may contribute to the distinction of obligatory from facultative phenomena during transformation. PMID- 2569256 TI - [Coronary bypass surgery using the internal mammary artery: results and 2-year clinical follow-up in Belgian patients]. AB - During the years 1984-1985, 867 coronary bypass operations were performed. The internal mammary artery was used in 471 patients (54.3%). Hospital mortality was 1.3%. Complications were mainly cardiac and respiratory. In the 160 Belgian patients a clinical follow-up study was performed. Mean follow-up was 28.9 +/- 7.3 months; 80% of patients had no symptoms; 6 patients (4%) underwent recatheterization because of new symptoms. In only one patient was the mammary graft occluded. During the follow-up period 5 patients died; this represents a late mortality of 3.4%. The advantages and disadvantages of venous grafts versus bypasses using the left internal mammary artery are discussed. In coronary artery bypass grafting the internal mammary artery should be used preferentially because of its resistance to atherosclerosis and excellent long-term patency. PMID- 2569255 TI - Action of lead on neurotransmission in rats. AB - 1. The neurotoxic effect of lead on the catecholaminergic and cholinergic nervous systems has been investigated using a rat model of lead exposure. 2. This model of lead exposure resulted in significant quantities of lead accumulating in the blood, brain and femur of the lead-exposed animals. 3. The biochemical effect of lead on brain neurochemistry was dependent on the degree and duration of lead exposure. However, the data points to a selective action of lead, with the midbrain and diencephalon being prime targets while very few lead-related alterations were observed in the cerebellum or the telencephalon. 4. Within the catecholaminergic nervous system, lead exposure resulted in alterations in the concentrations of the transmitters, noradrenaline and dopamine, in addition to changes in the activities of the enzymes tyrosine hydroxylase and phenylethanolamine-N-methyl transferase. The activity of the cholinergic biosynthetic enzyme, choline acetyltransferase was also noted to be altered by lead exposure. PMID- 2569257 TI - Immunohistochemical demonstration of neuroactive substances in the inner ear of rat and guinea pig. AB - The presence and localization of different neuropeptides and other putative neurotransmitters or -modulators were examined by immunohistochemistry in the cochleovestibular end organs and in neurons innervating them in rats and guinea pigs. In the organ of Corti neural elements beneath inner hair cells showed immunoreactivity for enkephalin (ENK), calcitonin gene-related peptide (CGRP), L glutamate decarboxylase (GAD), substance P (SP) and tyrosine hydroxylase (TH). Nerve chalices of type I vestibular hair cells contained SP and GAD, but not consistently. SP was only occasionally observed in neuronal cell bodies of the 8th cranial nerve but fine fibers with different neuroactive substances were seen in the nerve trunk in the following relative numbers: TH greater than SP greater than CGRP greater than ENK. The present data demonstrate the presence of several different neuroactive substances in the rat and guinea pig inner ear suggesting a multiplicity of neurotransmitters or -modulators in this system. PMID- 2569258 TI - Developmental changes in the chemosensitivity of rat brain synaptoneurosomes to excitatory amino acids, estimated by inositol phosphate formation. AB - The evolution of excitatory amino acids-(EAA) stimulated inositol phosphates (IPs) turnover during postnatal development was investigated in synaptoneurosomes prepared from rat forebrains. The two main EAA agonists which induce the IPs synthesis were quisqualate (QA) and N-methyl-D-aspartate (NMDA). The QA and NMDA stimulations of IPs formation present a particular developmental pattern, characterized by an active phase during rat synaptogenesis. The QA-evoked IPs accumulation peaked in synaptoneurosomes prepared from 8-day-old rat forebrains while that evoked by NMDA peaked in synaptoneurosomes from 12-day-old rats. These two developmental patterns are specific of the EAA agonists since the other various neuroactive substances tested (carbachol (Carb), noradrenaline, and high concentrations of potassium) induced an IPs accumulation, which increases during development and reaches a maximum in synaptoneurosomes of adult animals. Aging leads to a decrease in the capability of EAAs and muscarinic agonists to stimulate IPs formation in synaptoneurosomes, whereas the stimulation of IPs turnover by noradrenaline remains constant. Taken together, these results suggest that EAAs play a key role during brain development by sequentially activating two receptor subtypes, a new QA receptor, and a NMDA receptor, linked to the phosphoinositide metabolism. They may also indicate that these EAA-induced IPs responses are related to neuronal plastic events, the amplitude of which decreases with aging. PMID- 2569259 TI - Spatial distribution of glutamate, taurine and GABA in teleosts and mammals retina: in vivo and in vitro study. AB - Teleost and amphibian retina grows throughout life at the circumferential margin distal to the center. Retinas of two teleosts, goldfish and Eugerres plumieri; and two mammals, rat and rabbit were dissected into concentric regions; center and periphery; or center, intermediate and periphery. The concentration of glutamic acid, taurine, and gamma-aminobutyric acid was determined in these areas. A non-significant difference was found between the center and periphery of retinas dissected into two regions. By dissecting the teleost retina in three concentric regions a progressive increase of gamma-aminobutyric acid and taurine for center to periphery was observed, which was statistically significant for taurine. Rat and rabbit retina did not present this difference. The concentration of these amino acids was also determined in two concentric regions of goldfish retina 24 hr after crush of the optic nerve. Taurine content significantly increased in the center of the regenerating retina, but not in the periphery. Outgrowth was measured in explants prepared from central and peripheral retina in the presence of exogenous taurine. The amino acid produced an increase of the nerve growth index of central explants, but inhibited the growth from peripheral explants. This report presents a correlation between taurine concentration in a region of the retina that could be considered as embryonic tissue, suggesting that this amino acid plays a role in the formation of new cells. Moreover, the results from post-crush and cultured retina, with respect to taurine concentration or regenerating effect, suggest a saturation of the taurine enriched zone. PMID- 2569260 TI - Marine envenomations and aquatic dermatology. AB - Jellyfish stings are usually mild except those caused by species in the South Pacific. The box jellyfish can produce a severe cardiorespiratory insult. The sting of the Portuguese man-of-war is more potent than that of the common jellyfish. The Indo-Pacific area is the source of the most venomous bony fish. Many injuries can be avoided by wearing shoes when walking in shallow water or tide pools. Aquatic-related skin infections may involve unusual organisms. Swimmer's itch, a disease of freshwater bathing, is caused by cercariae. Seabather's eruption produces a rash in swimsuit-covered areas; the etiology is not clear. PMID- 2569261 TI - Effect of aspirin on myocardial ischemia. PMID- 2569262 TI - Pharmacologic profile of urapidil. AB - Urapidil, a phenylpiperazine-substituted derivative of uracil, is a selective alpha 1-adrenoceptor antagonist and therefore decreases blood pressure and peripheral resistance. Apart from its alpha 1-adrenoceptor-blocking potency, which is somewhat less selective than that of prazosin, urapidil appeared to display substantial central hypotensive activity. Initially, this central mechanism was assumed to be the same as that of clonidine, i.e., agonistic activity toward central alpha 2 adrenoceptors. This view, however, appeared to be incorrect for the following reasons: (1) In radioligand binding studies urapidil did not show any affinity for alpha 2 adrenoceptors; and (2) the central hypotensive activity of urapidil, injected into feline vertebral arteries remained unaffected by the alpha 2-adrenoceptor-blocking agents yohimbine and rauwolscine, which are known to suppress the central hypotensive effect of clonidine. The central hypotensive effect of urapidil so far remains unknown in detail, although a few (but not all) experimental arguments would plead for the 5 hydroxytryptamine (5-HT 1A)-agonistic potency of urapidil as an explanation of its central activity. The central hypotensive mechanism of urapidil will be discussed in more detail later in this issue. A presynaptic alpha 2-agonistic activity can only be demonstrated in certain models but not in the classic pithed rat preparation. It is unlikely to play a relevant role in urapidil's hypotensive activity, and also because of the poor affinity of the drug for alpha 2 adrenoceptors. Finally, a weak beta 1-adrenoceptor activity of urapidil can be demonstrated, but this mechanism is unlikely to contribute significantly to the drug's hypotensive activity. PMID- 2569263 TI - Urapidil, a multiple-action alpha-blocking drug. AB - Investigations in animals indicate that urapidil has a number of actions that may be relevant to its antihypertensive effect. It has an alpha 1-blocking action, a weak beta 1-blocking effect, an interaction with a serotonin receptor and a central depression of sympathetic tone. Urapidil is well absorbed orally with a bioavailability of about 70% and a time to peak concentration of about 4 hours after a sustained release capsule. It is metabolized in the liver at a half-life of 4.7 hours. Peripheral alpha 1-blocking activity has been demonstrated in humans. A shift to the right in the dose-response curve to phenylephrine has been found after urapidil, whereas responses to angiotensin are not affected. Evidence for beta 1-blocking activity is marginal. Urapidil does not inhibit the exercise increase in heart rate. Some investigators have suggested a possible inhibition of isoprenaline tachycardia; others have found no evidence. There is some evidence suggestive of a central action of urapidil in humans as lower single doses result in a decrease in blood pressure and an increase in heart rate. With higher doses the hypotensive effect continues but the tachycardia no longer occurs. However, urapidil has been reported to increase noradrenaline levels, although there has been a report with a high dose reducing vanillylmandelic acid excretion. Evidence for changes in renin is inconsistent. Hemodynamic studies have revealed findings that are compatible with peripheral alpha 1 blockade. After intravenous administration, peripheral resistance is reduced along with arterial pressure, and cardiac output is increased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569264 TI - Conclusions on the position of urapidil. PMID- 2569265 TI - Involvement of brain 5-HT1A receptors in the hypotensive response to urapidil. AB - Stimulation of serotonin-1A (5-hydroxytryptamine) (5-HT1A) receptors in the brain stem has been suggested to contribute to the antihypertensive action of the alpha 1-adrenoceptor antagonist urapidil. This hypothesis was tested by analyzing the influence of the 5-HT1A receptor antagonist spiroxatrine on the hypotensive responses to urapidil and the 5-HT1A receptor agonist 8-hydroxy-2-(di-n propylamino) tetralin (8-OH-DPAT). Chloralose/urethane-anesthetized cats underwent thoracotomy and were artificially ventilated. Blood pressure was monitored in the femoral artery. Urapidil (0.01 to 10 mumol/kg) or 8-OH-DPAT (3 to 30 nmol/kg) was injected into a femoral vein and the maximal hypotensive response recorded. A dose-response test with both drugs was performed before and after administration of spiroxatrine (3 and 10 nmol/kg); the latter was given through the vertebral artery, thus delivering the antagonist to the brain stem. Blood pressure was dose-dependently reduced by urapidil and 8-OH-DPAT after intravenous injection. Central administration of spiroxatrine through the vertebral artery shifted the dose-response curves of both drugs markedly and in a dose-dependent manner to the right, while the hypotensive response to the peripheral vasodilator nitroglycerin remained unchanged. The results suggest that the hypotensive response after peripheral administration of urapidil is mediated in part by stimulation of brain 5-HT1A receptors and this effect on central cardiovascular regulation is additive to the blood pressure reduction resulting from peripheral alpha-adrenoceptor blockade. PMID- 2569266 TI - Simultaneous determination of absolute total lymphocyte and CD4+ lymphocyte levels in peripheral blood by flow cytometry. AB - A distinguishing feature of the Spectrum III flow cytometer is its capacity to analyze a constant volume of cell suspension. The authors have capitalized on this feature to directly and simultaneously measure the absolute numbers of all lymphocytes and CD4+ lymphocytes per microliter of peripheral blood. The study group consisted of 42 hospital patients, 12 former blood donors seropositive for human immunodeficiency virus (HIV), and 12 HIV-seronegative donors. Regression analysis revealed a highly significant correlation (r = 0.97) between Spectrum lymphocyte counts and lymphocyte counts determined by a Coulter Counter S-Plus IV. Similarly, Spectrum CD4 cell counts were significantly correlated (r = 0.98) with CD4 cell counts calculated from the Coulter lymphocyte counts and % CD4+ lymphocytes determined by flow cytometry. These findings indicate that the absolute numbers of lymphocytes and subsets of lymphocytes in peripheral blood can be rapidly and simultaneously measured by flow cytometry. Such an assay should prove useful in studies of HIV infection, where total lymphocyte and CD4 cell levels are important parameters for clinical staging and assessing responses to treatment. PMID- 2569267 TI - Hormonal therapy for cryptorchidism with a combination of human chorionic gonadotropin and follicle-stimulating hormone. Success and relapse rate. AB - We treated 163 patients (Tanner stage I), aged 1 to 11 years, with cryptorchidism with a combination of 500 to 2000 IU of human chorionic gonadotropin divided into two intramuscular injections and given weekly and 75 IU of follicle-stimulating hormone once a week for 6 weeks. One hundred twelve patients had unilateral cryptorchidism. Response to therapy, which is descent of testes into scrotum, by age group was as follows: 2 (13.3%) of 15 patients aged 1 to 2 years; 8 (29.6%) of 27 patients aged 3 to 4 years; 13 (38.2%) of 34 patients aged 5 to 6 years; and 18 (50%) of 36 patients aged 7 to 11 years. Fifty-one patients had bilateral cryptorchidism. Response by age group was as follows: 1 (16.6%) of 6 patients aged 1 to 2 years; 3 (27.2%) of 11 patients aged 3 to 4 years; 6 (37.5%) of 16 (plus unilateral descent in 1 patient) aged 5 to 6 years; and 10 (55.5%) of 18 patients aged 7 to 11 years. The results are comparable with those obtained with human chorionic gonadotropin treatment alone. A relapse rate of 9.7% after 18 months of follow-up seemed to be lower compared with those reported with treatment with gonadotropin-releasing hormone or treatment with human chorionic gonadotropin alone. PMID- 2569268 TI - DNA rearrangement and restriction fragment length polymorphism within the first BCR intron in Philadelphia-positive acute leukemia. AB - In some patients with Philadelphia (Ph)-chromosome positive acute lymphoblastic leukemias, breakpoints on chromosome 22 are reported to occur within the first BCR intron. To analyze the breakpoints in chromosome 22 of Ph-positive acute leukemia patients without rearrangement of the 5.8 kb bcr, we cloned the 3' part of the first BCR intron using a synthetic DNA probe. During the course of study, we mapped the region of the deletion/insertion of 1 kb that causes a restriction fragment length polymorphism (RFLP) and found a racial difference in the frequencies of the alleles giving rise to this RFLP. Analyses of the patients' samples indicated that breakpoints were located within the 8.5 kb EcoRl fragment of the first BCR intron in two of five Ph-positive acute leukemia patients. The data, together with the previous reports, indicate that breakpoints within this approximately 50 kb intron are widely scattered, in contrast to those confined within the 5.8 kb bcr in chronic myelogenous leukemias. PMID- 2569269 TI - Linkage analysis of families with hereditary retinoblastoma: nonpenetrance of mutation, revealed by combined use of markers within and flanking the RB1 gene. AB - Nonpenetrance of the inherited mutation responsible for retinoblastoma has been reported. By DNA analysis in families with hereditary retinoblastoma, it is possible to identify healthy individuals in whom the mutation is nonpenetrant. This requires the use of DNA markers both within and flanking the retinoblastoma gene. We have analyzed the segregation of several markers in 19 families (69 meioses) with hereditary retinoblastoma. In two families a carrier was identified who showed nonpenetrance of the mutation predisposing to retinoblastoma. The intragenic markers were informative in 15 pedigrees. The use of flanking markers from the same chromosomal region caused an increase of the number of informative families to 18. No crossing-over within the gene was observed. In one family an inherited deletion involving one of the RB1 alleles was detected. Our findings emphasize the use of a combination of both intragenic and flanking markers to obtain both the highest reliability of carrier detection in families with hereditary retinoblastoma and an accurate estimate of the frequency of nonpenetrance. PMID- 2569270 TI - Linkage analysis of families with fragile-X mental retardation, using a novel RFLP marker (DXS 304). AB - A new polymorphic DNA marker U6.2, defining the locus DXS304, was recently isolated and mapped to the Xq27 region of the X chromosome. In the previous communication we describe a linkage study encompassing 16 fragile-X families and using U6.2 and five previously described polymorphic markers at Xq26-q28. One recombination event was observed between DXS304 and the fragile-X locus in 36 informative meioses. Combined with information from other reports, our results suggest the following order of the examined loci on Xq: cen-F9-DXS105-DXS98-FRAXA DXS304-(DXS52-F8 -DXS15). The locus DXS304 is closely linked to FRAXA, giving a peak lod score of 5.86 at a corresponding recombination fraction of .00. On the basis of the present results, it is apparent that U6.2 is a useful probe for carrier and prenatal diagnosis in fragile-X families. PMID- 2569271 TI - Polymorphic DNA haplotypes at the phenylalanine hydroxylase (PAH) locus in European families with phenylketonuria (PKU). AB - DNA haplotype data from the phenylalanine hydroxylase (PAH) locus are available from a number of European populations as a result of RFLP testing for genetic counseling in families with phenylketonuria (PKU). We have analyzed data from Hungary and Czechoslovakia together with published data from five additional countries--Denmark, Switzerland, Scotland, Germany, and France--representing a broad geographic and ethnographic range. The data include 686 complete chromosomal haplotypes for eight RFLP sites assayed in 202 unrelated Caucasian families with PKU. Forty-six distinct RFLP haplotypes have been observed to date, 10 unique to PKU-bearing chromosomes, 12 unique to non-PKU chromosomes, and the remainder found in association with both types. Despite the large number of haplotypes observed (still much less than the theoretical maximum of 384), five haplotypes alone account for more than 76% of normal European chromosomes and four haplotypes alone account for more than 80% of PKU-bearing chromosomes. We evaluated the distribution of haplotypes and alleles within these populations and calculated pairwise disequilibrium values between RFLP sites and between these sites and a hypothetical PKU "locus." These are statistically significant differences between European populations in the frequencies of non-PKU chromosomal haplotypes (P = .025) and PKU chromosomal haplotypes (P much less than .001). Haplotype frequencies of the PKU and non-PKU chromosomes also differ significantly (P much less than .001. Disequilibrium values are consistent with the PAH physical map and support the molecular evidence for multiple, independent PKU mutations in Caucasians. However, the data do not support a single geographic origin for these mutations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569272 TI - Polymorphic DNA haplotypes at the phenylalanine hydroxylase (PAH) locus in Asian families with phenylketonuria (PKU). AB - DNA polymorphisms at the phenylalanine hydroxylase (PAH) locus have proved highly effective in linkage diagnosis of phenylketonuria (PKU) in Caucasian families. More than 10 RFLP sites have been reported within the PAH structural locus in Caucasians. With information from affected and unaffected offspring in PKU families it is often possible to reconstruct complete RFLP haplotypes in parents and to use these haplotypes to follow the segregation of PKU within families and to determine the distribution of PKU chromosomes within populations. To establish the utility of these RFLPs in characterizing Asian families with PKU, we typed eight DNA sites in 21 Chinese families and 12 Japanese families with classical PKU. The eight RFLPs were chosen for their informativeness in Caucasians. From these families we reconstructed a total of 91 complete PAH haplotypes, 44 from non-PKU chromosomes and 47 from PKU-bearing chromosomes. Although all eight marker sites are polymorphic in both Chinese and Japanese, there is much less haplotypic variation in Asians than in Caucasians. In particular, one haplotype alone, haplotype 4, accounts for more than 77% of non-PKU chromosomes and for more than 80% of PKU-bearing chromosomes. Haplotype 4 is also relatively common in Caucasians. The next most common Asian haplotype is 10 times less frequent than haplotype 4. By contrast, in many Caucasian populations the sum of the frequencies of the five most common haplotypes is still less than 80%, and several of the most common haplotypes are equally frequent. Even though the extent of haplotypic variation in Asians is severely limited, the few haplotypes that are found often differ at a number of RFLP sites.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569273 TI - Tonic pupillary reaction after epidemic nephropathy and transient myopia. PMID- 2569275 TI - Phenytoin-induced resistance to vecuronium. PMID- 2569274 TI - Beta blockade to decrease silent myocardial ischemia during peripheral vascular surgery. AB - The incidence and duration of intraoperative silent myocardial ischemia have been shown to be significantly correlated with the incidence of perioperative myocardial infarction in patients undergoing peripheral vascular surgery. To assess the effectiveness of intraoperative beta blockade in limiting such silent myocardial ischemia, a group of 48 patients was treated with oral metoprolol immediately prior to peripheral vascular surgery. The total duration of intraoperative silent myocardial ischemia, the percentage of intraoperative time silent myocardial ischemia was present, the number of intraoperative episodes of silent myocardial ischemia, and the intraoperative heart rate in the treated patients were compared with those in 152 similar but untreated peripheral vascular surgery patients. The patients treated with oral metoprolol had significantly less intraoperative silent ischemia with respect to relative duration and frequency of episodes, a significantly lower intraoperative heart rate, and less intraoperative silent myocardial ischemia in terms of total absolute duration. These results suggest that beta-adrenergic activation may play a major role in the pathogenesis of silent myocardial ischemia during peripheral vascular surgery. PMID- 2569276 TI - High-performance liquid chromatographic determination of bromazepam in human plasma. AB - An assay using high-performance liquid chromatography has been developed for the determination of bromazepam in plasma. After a single-step extraction from basified samples with dichloromethane, using decarboxyloflazepate as an internal standard, samples were analysed using a reversed-phase Nova Pak 5-microns column with a mobile phase of methanol - phosphate buffer (60 + 40) adjusted to pH 7.6. The drugs were detected at 239 nm and the limit of detection was found to be 3 micrograms l-1 for bromazepam. The method is simple, rapid and sensitive and permits bromazepam levels in clinical and pharmacokinetic studies to be monitored. PMID- 2569277 TI - Chromogranin- and somatostatin-containing neuroendocrine cells in the porcine uterus. An immunocytochemical study. AB - Endocrine cells scattered in organic mucosae were defined "Neuroendocrine" (NE) cells because they constitute a section of the Diffuse Neuroendocrine System (DNES). Such cells have never been evidentiated in the normal endometrium. By means of histochemical and immunohistochemical techniques, NE cells, some of which contain the hormone somatostatin, are described in the glandular epithelium of the uterine horn in non-pregnant, non-castrated, young and adult sows. As is known, the uterine horn is the organ of pregnancy in the pig. The localization, distribution and morphology of the uterine NE and somatostatin-containing cells are reported and the importance of their function, in the pregnant and non pregnant porcine uterus, is discussed. PMID- 2569278 TI - The effect of H2-receptor antagonist premedication on the duration of vecuronium induced neuromuscular blockade in postpartum patients. AB - The clinical duration of vecuronium was measured in two groups of postpartum patients undergoing elective tubal ligation. Ten patients received no premedication and ten others ranitidine 150 mg orally the morning of surgery. The mean duration of action of vecuronium was 57.2 +/- 9.9 min in the unmedicated patients and 54.0 +/- 12.9 min in the ranitidine treated patients. These values were significantly greater than the mean value for nonpregnant control patients (35.3 +/- 8.4 min) but indistinguishable from the mean value for cimetidine pretreated patients (63.0 +/- 17.6 min) reported previously. The combined results of the previous and present studies provide convincing evidence that the clinical duration of vecuronium-induced neuromuscular blockade is significantly longer in the postpartum patient and independent of cimetidine or ranitidine pretreatment. PMID- 2569280 TI - The effect of neuromuscular blockade with vecuronium on hemodynamic responses to noxious stimuli in the rat. AB - The effect of neuromuscular blockade with vecuronium on the hemodynamic responses to a noxious stimulus was investigated in male Sprague-Dawley rats. The rats were anesthetized with either halothane (group 1, n = 10), or isoflurane (group 2, n = 10). The maximum values for heart rate and mean arterial pressure during the noxious stimulus (base-tail clamp) were measured, and the maximum changes in these values (maximum minus prestimulation) were calculated. The responses were measured at two different anesthetic concentrations (0.6 X MAC, 0.75 X MAC), before and after vecuronium 1.0 mg.kg-1 iv. It was found that neuromuscular blockade with vecuronium did not reduce any of the hemodynamic responses measured, at either anesthetic concentration, in either the halothane or the isoflurane group. However, increasing the anesthetic concentration from 0.6 X MAC to 0.75 X MAC produced statistically significant (P less than 0.01) reductions in several of the responses measured. The inability of vecuronium to reduce hemodynamic responses to noxious stimuli in this study suggests that neuromuscular blockade does not alter anesthetic depth in the rat. A knowledge of this "absence of effect" may be important for investigators who need to induce muscle relaxation in laboratory animals prior to examining the effect of anesthetic agents on hemodynamic responses to noxious stimuli. The results also question the ability of neuromuscular blockade to reduce anesthetic requirement, and support the view that neuromuscular blockade does not contribute to the anesthetic state. PMID- 2569279 TI - Onset and duration of neuromuscular blockade following high-dose vecuronium administration. AB - To determine the onset time and duration of high doses of vecuronium, 40 ASA Physical Status 1 and 2 patients were randomly assigned to receive either 100, 200, 300, or 400 micrograms/kg of vecuronium bromide for muscle relaxation during elective general surgery. Neuromuscular blockade was continuously quantitated by recording the electromyographic response to stimulation of the ulnar nerve train of-four. The rate of onset of neuromuscular blockade, endotracheal intubating conditions, duration of neuromuscular blockade, and hemodynamic effects of vecuronium at each dose were evaluated and compared. The time from vecuronium administration to complete abolition of twitch tension (T1 = 0%) decreased from 208 +/- 41 to 106 +/- 35 s as the vecuronium dose was increased from 100 to 400 micrograms/kg (P less than 0.01). Corresponding times to endotracheal intubation (T1 less than 20%) also decreased from 183 +/- 24 to 96 +/- 31 s with increasing doses (P less than 0.01). Recovery time (T1 = 25%) increased from 37 +/- 13 to 138 +/- 24 min with increasing doses (P less than 0.01). No significant hemodynamic differences between the four groups were observed. Endotracheal intubating conditions were good or excellent in all patients. High doses of vecuronium may, therefore, be a useful alternative to succinylcholine when a rapid onset of neuromuscular blockade is required. PMID- 2569281 TI - Alterations of alpha 1 and alpha 2 adrenoceptor-mediated pressor responses by halothane and isoflurane anesthesia. AB - The mechanism by which halothane and isoflurane interfere with catecholamine mediated vasoconstriction was investigated, utilizing selective agonists of postjunctional alpha1 and alpha2 adrenoceptors in chronically instrumented dogs. After ganglionic, cholinergic, and beta adrenergic blockade, dose responses to phenylephrine (0.3-1.2 micrograms/kg, iv), a selective alpha1 adrenoceptor agonist, and azepexole [B-HT 933] (5-20 micrograms/kg, iv), a selective alpha2 adrenoceptor agonist, were obtained in conscious dogs. Each dog was subsequently anesthetized with either halothane (1.7%) or isoflurane (2%) in oxygen in equihypotensive concentrations. After a 1 h equilibration period, the dose response curves were repeated. Twenty experiments in ten chronically instrumented dogs were completed. Halothane and isoflurane produced significant (P less than 0.05) attenuation of both the increase in systolic and diastolic arterial pressure after bolus administration of all doses of phenylephrine and azepexole. No specific selectivity of either volatile anesthetic for alpha1 or alpha2 mediated pressor responses was found. Therefore, in chronically instrumented dogs, alpha1- and alpha2-mediated pressor responses were similarly influenced by halothane and isoflurane. The present results suggest that both halothane and isoflurane act as functional antagonists to alpha adrenergic mediated vasoconstriction. PMID- 2569282 TI - Dexmedetomidine, acting through central alpha-2 adrenoceptors, prevents opiate induced muscle rigidity in the rat. AB - The highly-selective alpha-2 adrenergic agonist dexmedetomidine (D-MED) is capable of inducing muscle flaccidity and anesthesia in rats and dogs. Intense generalized muscle rigidity is an undesirable side effect of potent opiate agonists. Although the neurochemistry of opiate-induced rigidity has yet to be fully elucidated, recent work suggests a role for a central adrenergic mechanism. In the present study, the authors determined if treatment with D-MED prevents the muscle rigidity caused by high-dose alfentanil anesthesia in the rat. Animals (n = 42) were treated intraperitoneally with one of the following six regimens: 1) L MED (the inactive L-isomer of medetomidine), 30 micrograms/kg; 2) D-MED, 10 micrograms/kg; 3) D-MED, 30 micrograms/kg; 4) D-MED [30 micrograms/kg] and the central-acting alpha-2 antagonist, idazoxan [10 mg/kg]; 5) D-MED [30 micrograms/kg] and the peripheral-acting alpha-2 antagonist DG-5128 [10 mg/kg], or; 6) saline. Baseline electromyographic activity was recorded from the gastrocnemius muscle before and after drug treatment. Each rat was then injected with alfentanil (ALF, 0.5 mg/kg sc). ALF injection resulted in a marked increase in hindlimb EMG activity in the L-MED treatment group which was indistinguishable from that seen in animals treated with saline. In contrast, D-MED prevented alfentanil-induced muscle rigidity in a dose-dependent fashion. The small EMG values obtained in the high-dose D-MED group were comparable with those recorded in earlier studies from control animals not given any opiate. The high-dose D-MED animals were flaccid, akinetic, and lacked a startle response during the entire experimental period.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569283 TI - Esmolol for perioperative management of thyrotoxic goiter. PMID- 2569284 TI - Restriction fragment length polymorphisms associated with growth hormone and prolactin genes in Holstein bulls: evidence for a novel growth hormone allele. AB - Sperm DNA isolated from sons of three extensively used US Holstein bulls was screened for differences associated with the primary gene structure of the bovine growth hormone (bGH) and prolactin (bPrl) genes. Southern blot analysis of DNA digested with 10 restriction enzymes revealed that offspring from two of the three bull families exhibited polymorphisms around the bGH and bPrl genes. Restriction fragment length polymorphisms (RFLPs) around the bGH gene were detected with five enzymes, whereas three enzymes revealed RFLPs around the bPrl gene. At least three structural differences were predicted around the bGH gene. The most common variant hybridization pattern appeared to involve an insertion/deletion located downstream of the conserved 3' EcoRI site. The presence of RFLPs in the genes coding for these pituitary hormones within a familial line may provide the basis for genetic markers associated with lactation and mammary development. PMID- 2569285 TI - Restriction fragment length polymorphism analysis of major histocompatibility complex class II genes from inbred chicken lines. AB - High molecular weight DNA was extracted from sperm from chickens of 14 inbred lines. The DNA was digested with each of four restriction enzymes (Pvu II, Hind III, Bgl II, and Bam HI), electrophoresed for 18 or 45 h, blotted onto nitrocellulose, and hybridized to a chicken major histocompatibility complex (MHC, B complex) class II beta-chain probe (beta 2-exon specific). Restriction fragment length polymorphisms (RFLPs) were found with each of the restriction enzymes used. Birds with the same B haplotype always showed the same RFLP pattern; however, some birds of different B haplotypes also shared the same RFLP pattern. To test for the Mendelian inheritance of the RFLP patterns, the F2 progeny of an informative cross were analysed. The RFLP patterns corresponded with the serologically determined B haplotypes of the F2 birds, thereby showing the Mendelian inheritance of the polymorphic bands. PMID- 2569286 TI - Genentech, Inc, and symposium sponsorship. PMID- 2569287 TI - Taxol: a unique antineoplastic agent with significant activity in advanced ovarian epithelial neoplasms. AB - STUDY OBJECTIVE: To assess the activity of taxol in patients with advanced, progressive, and drug-refractory ovarian cancer and to delineate more clearly the toxicity of taxol in this patient population. DESIGN: Nonrandomized, prospective phase II trial. PATIENTS: Forty-seven patients with drug-refractory epithelial ovarian cancer who had one or more lesions measurable in perpendicular diameters. Of these patients, 45 were evaluable for toxicity and 40 were evaluable for response. INTERVENTIONS: PATIENTS were treated every 22 days with varying doses of taxol (110 to 250 mg/m2 body surface) given as a 24-hour infusion with subsequent doses based on adverse effects. A premedication regimen was used to avoid acute hypersensitivity reactions. MEASUREMENTS AND MAIN RESULTS: Twelve patients (30%; CI, 16% to 44%) responded to taxol for periods lasting from 3 to 15 months. The dose-limiting toxicity was myelosuppression with leukocytes affected more severely and commonly than thrombocytes or reticulocytes. Leukopenia was usually brief in duration but was associated with sepsis in 3 cases (2 fatal). Other adverse effects included myalgias, arthralgias, alopecia, diarrhea, nausea, vomiting, mucositis, and peripheral neuropathy. Rare cases of cardiac and central neurotoxicity were also noted. CONCLUSIONS: Taxol is an active agent in drug-refractory ovarian cancer and deserves further study in combination with other active drugs in previously untreated patients with advanced ovarian cancer. PMID- 2569288 TI - [Angina without involvement of large coronary trunks and precapillary pulmonary hypertension in a patient with periarteritis nodosa]. AB - A patient affected with periarteritis nodosa and angina, presented a normal coronary angiography of the coronary trunks and an alteration of the left ventricular systolic function, improved by nicardipine, as well as a pre capillary pulmonary hypertension with isolated, right cardiac repercussions, without pulmonary fibrosis nor thromboembolic disease. In addition, he presented three transient cerebral ischaemic accidents, with different localizations, which could not be related to a right aneurysm of the sylvian artery. Isolated precapillary pulmonary hypertension, without pulmonary fibrosis, which is not described in the course of periarteritis nodosa, seems related to an arteriolar involvement which is the known substratum of other visceral involvement in periarteritis nodosa. Improvement of the left ventricular function with nicardipine, administered acutely, suggest a partially reversible disorder of the coronary microcirculation. PMID- 2569289 TI - The anonymous PAS45 probe detects RFLPs in 13q31. AB - An anonymous DNA probe PAS45 was isolated. This probe detects an RFLP with two alleles 1 and 2 at the same locus, with the different restriction enzymes (Bg1II, EcoRI, HindIII, PstI, MspI, XbaI). The observed polymorphism is explained by a chromosome rearrangement involving these enzyme cleavage sites. The frequency of alleles 1 and 2 was 0.875 and 0.125, respectively, in a sample of 48 unrelated individuals in France. Codominant inheritance of alleles 1 and 2 was demonstrated in 13 families with 30 offspring. The PAS45 probe was localized on chromosome 13 by somatic cell hybrid analysis and on 13q31 by in situ hybridization. The rearrangement on 13q31 is present in one out of four healthy individuals in France. PMID- 2569290 TI - RFLPs tightly linked with cystic fibrosis: value of probes at the D7S23 locus in prenatal diagnosis. AB - In a 1:4 risk family, the usefulness of probes at the D7S23 locus for prenatal diagnosis of cystic fibrosis is discussed by comparison with probes at the MET, D7S8, and D7S18 loci that did not allow accuracy in this family. PMID- 2569292 TI - Measurement of constitutive L-pyrrolidonyl peptidase activity from Streptococcus and Enterococcus using tetrazotized 0-dianisidine. AB - The detection of L-pyrrolidonyl peptidase activity is extremely useful for the differentiation of Enterococcus species and Streptococcus pyogenes from other members of the family Streptococaceae. This analysis has generally been performed utilizing the hydrolyzable substrate L-pyrrolidonyl beta-naphthylamine. After the substrate was hydrolyzed, free beta-naphthylamine has been detected utilizing the reagent para-dimethylaminocinnamaldehyde. The cinnamaldehyde and naphthylamine reagents combined to form an orange color, much like the indole reaction. The use of the cinnamaldehyde reagent had several drawbacks however: color development was not sharp, and the reagent was difficult to produce, and it was not stable. A new indicator system employing tetrazotized 0-dianisidine was developed. An extremely deep burgundy colored complex resulted from the reaction between the new indicator and B-Naphthylamine. This diazo reagent showed excellent correlation with results obtained with para-dimethylaminocinnamaldehyde and yielded more objective, distinct endpoints. This inexpensive reagent can be utilized either in a liquid form or dried on paper discs. PMID- 2569291 TI - Organic nitrogen metabolism of phototrophic bacteria. AB - Recent reviews dealing with phototrophic bacteria are concerned with bioenergetics, nitrogen fixation and hydrogen metabolism, synthesis of the photosynthetic apparatus and phylogeny/taxonomy. The organic N-metabolism of these phylogenetically diverse bacteria has last been reviewed in 1978. However, amino acid utilization and biosynthesis, ammonia assimilation, purine and pyrimidine metabolism and biosynthesis of delta-aminolevulinic acid as precursor of bacteriochlorophylls and hemes are topics of vital importance. This review focuses on utilization of amino acids as N- and C/N-sources, the pathways of purine and pyrimidine degradation, novel aspects of amino acid biosynthesis (with emphasis on branched-chain amino acids and delta-aminolevulinic acid) and some aspects of ammonia assimilation and glutamate synthesis by purple bacteria, green sulfur bacteria and Chloroflexus aurantiacus. PMID- 2569293 TI - Evidence for the presence of a very high concentration of arylsulfatase A in the pig thyroid: identification of arylsulfatase A subunits as the two major glycoproteins in purified thyroid lysosomes. AB - In addition to their general function in cellular homeostasis, thyroid lysosomes play an essential role in the biosynthesis of thyroid hormones by cleaving the macromolecular prohormone, thyroglobulin. In the present work, we have attempted to determine whether the enzyme composition of thyroid lysosomes differs from that of lysosomes from other tissues. Lysosomal enzymes, cathepsin D, beta-D galactosidase, beta-D-glucosidase, alpha-D-mannosidase, alpha-L-fucosidase, hexosaminidase, and arylsulfatase A and B, were assayed in crude fractions from various pig tissues, heart, brain, liver, kidney, thyroid, adrenals, ovary, and spleen. It appeared that the specific activity of arylsulfatase A was at least 20 times higher in the thyroid than in most other tissues. Thyroid lysosomes purified by isopycnic centrifugation on Percoll gradients contained two major polypeptides with apparent molecular weights of 58,000 and 54,000 representing about 30% of the total protein. These polypeptides were glycosylated and were exclusively found in the intralysosomal soluble fraction obtained by osmotic pressure-dependent lysis. By fractionating intralysosomal soluble proteins by velocity sedimentation on sucrose gradients or gel permeation chromatography we identified a thyroid arylsulfatase A holoenzyme which corresponds to a 120,000 Mr species. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analyses of the gradient or column fractions showed that the 120-kDa protein peak with arylsulfatase A activity essentially contained the 58- and 54-kDa polypeptides in equivalent amounts. In conclusion, arylsulfatase A, a heterodimer of 120 kDa composed of two nonidentical subunits, is the major protein component of thyroid lysosomes. The superabundance of this protein in purified thyroid lysosomes is related to the very high specific activity of the enzyme in the thyroid as compared to other tissues. PMID- 2569294 TI - Stereospecificity of beta-adrenoceptor blockade and renal vasodilatation by tertatolol. AB - To investigate the stereospecificity of beta-adrenoceptor blockade and renal vasodilatation by tertatolol, the regional hemodynamic effects of the 2 stereo isomers of tertatolol (d-tertatolol and l-tertatolol) were compared in conscious spontaneously hypertensive rats. Furthermore, the effects of a combination of d tertatolol and dl-propranolol were studied. l-Tertatolol and the combination of beta-blockers lowered heart rate and caused an early increase and a late decrease of blood pressure. l-Tertatolol, but not the combination of dl-propranolol and d tertatolol, decreased renal resistance at 3-4 hr after injection. Injections of d tertatolol alone did not cause any effect. Since effects of l-tertatolol are fully comparable to effects previously observed with dl-tertatolol, and d tertatolol was devoid of any effects, both alone and in combination with dl propranolol, we conclude that both beta-adrenoceptor blockade and renal vasodilatation are stereospecific for the l-isomer of tertatolol. PMID- 2569295 TI - Sodium regulation of alpha 2-adrenoceptor-agonist interactions in renal membranes of young and old Dahl rats. AB - Systolic blood pressure, the affinity and density of the alpha 2-adrenoceptors, Na+ effect on alpha 2-adrenoceptors as measured by specific [3H]yohimbine binding, and Na+ regulation of alpha 2-adrenoceptor-agonist interactions were studied in 6 week and 9 month old inbred salt sensitive (S) and inbred salt resistant (R) rats. The systolic blood pressure of 6 week old S and R rats was not significantly different, while the systolic blood pressure of 9 month old S rats was higher than the age-matched R rats. The affinity of the alpha 2 adrenoceptors for yohimbine was the same in the 6 week old S and R rats but it was 2-fold less in the 9 month old S rats compared to the age-matched R rats. In the 6 week old rats, the number of alpha 2-adrenoceptors (Bmax) was significantly higher in the S rats than in the R rats; whereas, in the 9 month old rats, the alpha 2-adrenoceptors were 2.6 times higher in the S rats than the age-matched R rats. Na+, up to 80 mM, did not modify the number of alpha 2-adrenoceptors in the 6 week and 9 month old S and R rats. However, Na+ reduced the affinity of the alpha 2-adrenoceptors for epinephrine and this effect of Na+ was found to be the same in the 6 week and 9 month old S and R rats. These findings suggest that Na+ regulation of alpha 2-adrenoceptor-agonist interactions was not altered with age and it was found to be similar in prehypertensive rats and rats with severe hypertension; further, the resistance of R rats to salt-induced hypertension was not due to the absence of Na+ binding component in the alpha 2-adrenoceptor adenylate cyclase complex. PMID- 2569297 TI - Is in vivo dissociation between the antipropulsive and antidiarrheal properties of opioids in rats related to gut selectivity? AB - The antipropulsive activity of a series of opioids in the charcoal test was compared with their antidiarrheal activity in the castor oil test and their analgesic activity in the tail withdrawal test. The obtained antipropulsive/antidiarrheal potency ratios varied from 0.71 to greater than 552 [pethidine (oral ED50's in mg/kg: 21.5/30.2), fentanyl (0.77/0.49), dextromoramide (5.39/2.90), methadone (14.2/6.38), codeine (98.4/10.8), morphine (56.6/5.21), diphenoxylate (8.15/0.54), nufenoxole (74.7/1.72), difenoxin (7.10/0.16), loperamide oxide (greater than 160/0.34) and loperamide (greater than 160/0.29)]. The above ratios correlated with the gut selectivity of the compounds as defined by their analgesic/antidiarrheal potency ratios (r = 0.92, P less than 0.001). Furthermore, inhibition of propulsion was found to correlate with central analgesic activity (r = 0.93, P less than 0.001) but not with protection from diarrhea (r = 0.023, P greater than 0.05). Indeed, gut-selective opioids such as loperamide and loperamide oxide failed to affect propulsion up to doses more than 450 times their antidiarrheal doses. In contrast, alpha 2 adrenoceptor agonists delayed propulsion at doses comparable to their antidiarrheal doses [clonidine (0.085 vs 0.021), lidamidine (2.35 vs 1.66)] and anticholinergics inhibited propulsion even at doses many times below their antidiarrheal doses [atropine (0.26 vs 9.30), dexetimide (0.13 vs 5.03) and isopropamide (0.78 vs 74.6)]. The present results indicate that the in vivo inhibition of gastrointestinal propulsion by opioids in rats is mediated by a central action. Effects on intestinal fluid transport or, alternatively, on motility events distal to the ileocecal junction rather than effects on propulsion through the small intestine, seem to be the primary mechanism of antidiarrheal action of gut-selective opioids such as loperamide and loperamide oxide. PMID- 2569296 TI - Prejunctional beta-adrenoceptors in the isolated perfused rat kidney: a comparative study of the beta-blockers tertatolol and propranolol. AB - The interaction of the beta-blocker tertatolol with the prejunctional beta adrenoceptors of the rat renal vasculature was examined. In isolated Tyrode perfused rat kidneys isoproterenol (5 x 10(-7) M to 2 x 10(-6) M) slightly decreased the pressor responses to noradrenaline (0.5 nmol), but markedly increased those to electrical stimulation (6 Hz). Tertatolol at 3 x 10(-7) M inhibited both effects of isoproterenol. Electrical stimulation (6 Hz) caused an overflow of intact 3H-noradrenaline from kidneys previously labelled with the 3H transmitter; this stimulation-induced release was augmented by isoproterenol. Also this effect of isoproterenol was prevented by 3 x 10(-7) M of tertatolol. In different groups of rat kidneys labelled with 3H-noradrenaline, isoproterenol (5 x 10(-7) M) augmented the stimulation (1 Hz), induced 3H overflow and this increased transmitter output was concentration-dependently inhibited by tertatolol and propranolol; from the IC50 values it was calculated that tertatolol was 16 times more potent than propranolol. Even at concentrations up to 10(-5) M, neither tertatolol nor propranolol significantly altered the basal and stimulation-induced transmitter release from the rat kidneys. From the data we conclude that tertatolol is a powerful inhibitor of the prejunctional beta adrenoceptors in the renal circulation of the rat. Stimulation of these receptors by beta-agonists not only enhances transmitter release but also increases the pressor responses induced by electrical stimulation. PMID- 2569299 TI - Bromocriptine-induced hypothermia: D-2 receptor involvement. AB - Bromocriptine (2-16 mg/kg, i.p.) caused a dose-dependent fall in core body temperature of mice. The hypothermic effect of bromocriptine was decreased by pretreatment of the animals with sulpiride, but SCH 23390, methergoline, phenoxybenzamine, propranolol and atropine did not decrease the response. Administration of reserpine plus alpha-methyl-p-tyrosine prior to bromocriptine injection abolished hypothermia in mice. It is concluded that D-2 dopamine receptors may be involved in bromocriptine-induced hypothermia. PMID- 2569298 TI - Differential effects of nebivolol, atenolol and propranolol on heart rate and on bronchoconstrictor responses to histamine in the guinea-pig. AB - Changes in heart rate and in bronchomotor reactions to histamine as parameters for beta 1- and beta 2-adrenergic receptor blockade, respectively, were analyzed in anaesthetized, ventilated guinea-pigs after administration of propranolol, atenolol and nebivolol. Propranolol (0.04 to 0.63 mg/kg i.v., -15 min) produces a significant reduction of the resting heart rate (80.4 +/- 4.25 to 74.9 +/- 4.17% of premedication values; n = 8; p less than 0.05) and, at the same dose levels, a significant enhancement of the bronchoconstrictor responses to histamine (3 micrograms/kg i.v.) (129.1 +/- 6.3 to 167.7 +/- 11.2% of premedication values; n = 8; p less than 0.05). Atenolol (0.02 to 1.25 mg/kg i.v., -15 min) significantly reduces heart rates from 0.04 mg/kg on (79.4 +/- 2.6 to 69.3 +/- 3.2% of premedication value at 1.25 mg/kg; n = 8; p less than 0.05) and significantly enhances the pulmonary reaction to histamine from 0.63 mg/kg on (139.1 +/- 6.3% to 137 +/- 11.6% of premedication values at 1.25 mg/kg; n = 8; p less than 0.05), yielding a dissociation factor of 16 between the lowest cardiac active dose and the pulmonary active one. Nebivolol (0.08 to 2.5 mg/kg i.v., -15 min) significantly reduces heart rates from 0.125 mg/kg on (78 +/- 3.2% to 65.4 +/- 3.9% of premedication value at 2.5 mg/kg; n = 8; p less than 0.05) without significantly increasing pulmonary reactivity (132.2 +/- 5.4% of premedication values at 2.5 mg/kg; n = 8; p greater than 0.05), thus yielding a dissociation factor greater than 20 between cardiac-pulmonary active doses. The study demonstrates a substantial dissociation between the cardiac (beta 1 adrenoceptors) and pulmonary (beta 2-adrenoceptors) active doses of nebivolol and atenolol in comparison with propranolol in vivo. PMID- 2569300 TI - B-HT 920 antagonizes rat neophobia in the X-maze test: a comparative study with other drugs active on adrenergic and dopaminergic receptors. AB - Since there is compelling evidence to suggest a significant participation of central noradrenergic (NA) and dopaminergic (DA) pathways in the modulation of anxiety disorders, the possible tranquillizing effect of B-HT 920, considered to be a selective agonist of central DA autoreceptors at low doses, but also a stimulant of central alpha 2-receptors at high doses, was investigated by means of the X-maze test. For comparison, other drugs active on alpha-adrenoceptors and DA receptor subtypes were similarly tested. B-HT 920 at high doses (1 and 2 mg/kg) significantly antagonized the animals' spontaneous avoidance of open spaces (a sign indicative of anxiety), despite the reported sedative effects of the drug; it should be pointed out, however, that the latter were observable in our tests only at 1 mg/kg, a more pronounced sedation was the sole effect of a low dose (100 micrograms/kg) of B-HT 920. In our experimental conditions, clonidine (50 micrograms/kg) and yohimbine (5, but not 1 and 2.5 mg/kg) exerted anxiolytic and anxiogenic effects, respectively, in line with theoretical expectations and with results already published. DPI (100 and 200 micrograms/kg) (reported to be a selective agonist at DA receptors mediating neuronal inhibition and also active on alpha-adrenoceptors) and B-HT 958 (1 and 10 mg/kg) (a stimulant of DA autoreceptors but a blocker of alpha 2-adrenoceptors) were unable to modify the natural preference of rats for closed arms. Hypomotility was obtained with both doses of B-HT 958 and with the higher dose of DPI. PMID- 2569301 TI - Flow cytometric measurements of proliferation-associated nuclear antigen p105 and DNA content in non-Hodgkin's lymphomas. AB - Non-Hodgkin's lymphomas are characterized by heterogeneity in cell kinetics, which may be related to differential expression of proliferation-associated nuclear antigens. We have performed two-color flow cytometric quantitation of nuclear antigen p105 and DNA content in nuclear suspensions from 80 paraffin embedded lymphomas. High-grade lymphomas with diploid DNA histograms tended to express high levels of p105 in all phases of the cell cycle. Aneuploid populations, observed in 14 of 80 cases, showed high p105 expression compared with diploid cells of the same case, with the highest aneuploid:diploid p105 ratios in the high-grade group. We conclude that high-grade or DNA aneuploid lymphomas tend to have increased expression of p105; but considerable variations within each histologic subtype were observed, and the differences between histologic grades were not statistically significant. Further studies that correlate p105 expression and clinical outcome will help to determine the prognostic value of this marker. PMID- 2569302 TI - Synthesis and quantitative structure-activity relationships of analeptic agents related to dimefline. AB - Some new dimefline-type derivatives have been synthesized and their pharmacological activity, as well as their distribution coefficients have been determined. The distribution coefficients of a number of previously published analogue compounds have also been measured and the QSAR analysis of the whole set has been carried out. The results of such analysis allow to point out which factors are influencing the biological activity of this group of compounds. PMID- 2569303 TI - Ultrahistochemical demonstration of guanylate cyclase activity in the rat hippocampus. AB - Guanylate cyclase activity was demonstrated in the rat hippocampus at the electron microscopy level by a cerium precipitation technique. Enzymatic activity is localized in the granular cells of the dentate gyrus and in other, medium sized neurons. The reaction product was found on the cytoplasmic membrane, in the cytoplasm, on the surface of endoplasmic reticulum and in postsynaptic densities. Glial cell processes and small dendritic processes were also stained. Besides the neurons, glial cells contained strong enzymatic activity, especially the perivascular glia and protoplasmatic glial cells neighbouring to neurons. Only little activity was detected in pyramidal cells and no activity was found in the endothelium cells of capillaries. PMID- 2569305 TI - M&B 39890A, a novel sulphonamido-benzamide compound, inhibits insulin and glucagon secretion in vitro. PMID- 2569304 TI - Comparison between the activities of cationic amphiphilic drugs to affect phospholipid-membranes and to depress cardiac function. AB - The activity of cationic amphiphilic compounds to affect artificial phospholipid membranes was compared with the membrane-stabilizing cardiodepressant potency of the drugs. The twenty-one investigated catamphiphilic compounds belonged to various pharmacological groups including antiarrhythmic, local anaesthetic, beta blocking, antimalarial, and psychoactive drugs. The perturbing action of the drugs on phospholipid-membranes was evaluated by determining the drug-effect on the temperature of the phase-transition from the gel to the liquid crystalline state in liposomes of dipalmitoylphosphatidic acid by means of differential scanning calorimetry. The ability to interact with the polar headgroups of phospholipid-membranes was measured by recording the effects of the cationic compounds on the binding of 45Ca2+ to monomolecular layers of phosphatidylserine. The cardiodepressant action was observed in Langendorff-preparations of guinea pig hearts. The drug-effect on excitability was indicated by the elevation of the threshold-intensity of 50 Hz alternating current to induce ventricular arrhythmia. For the sake of comparison, the negative chronotropic and inotropic effects were evaluated. There was only a moderate correlation found between the activities of the drugs to reduce the transition temperature and to inhibit 45Ca2+ -binding (r = 0.69). This result probably reflects that both methods look upon different consequences of the drug-phospholipid interaction. The membrane stabilizing, anti-excitatory potency corresponded favourably with the ability of the drugs to affect the phospholipids. Almost 80% of the variance between the anti-excitatory potencies could be accounted for by the drug-phospholipid interactions. The negative chronotropic and inotropic effects accompanying the anti-excitatory actions were similar for most of the drugs. The results of the study are compatible with the hypothesis that the interaction with phospholipid membranes is a major determinant of the membrane-stabilizing cardiodepressant potency of cationic amphiphilic drugs. PMID- 2569306 TI - Detection of renal impairment in cases of chronic abuse of analgesics by administration of desamino-D-arginine vasopressin. AB - An early diagnosis of renal dysfunction is particularly important in patients with chronic abuse of analgesics. First morphological changes in cases of analgesic nephropathy are found in the renal medulla. Therefore, urinary osmolarity as well as the urinary excretion of cyclic adenosine monophosphate (AMP) and kallikrein were measured after application of desamino-D-arginine vasopressin (DDAVP) as parameters of the renal tubular function in 21 patients with normal creatinine clearance and a regular, excessive use of analgesics. The results were compared with those of 17 healthy volunteers and 8 patients with chronic renal insufficiency after analgesic nephropathy. In patients with analgesic nephropathy the urine osmolarity after 12 h of thirst was 367 +/- 28 mOsm/kg and did not increase after DDAVP. The results obtained from healthy volunteers were 781 +/- 39 mOsm/kg. Half of the patients with chronic misuse of analgesics had lower values of urine osmolarity after thirst and DDAVP compared to the reference area of healthy subjects. Similar results were found when the effect of DDAVP under conditions of water-loading was tested. The excretion of cyclic AMP as a second messenger of DDAVP and of kallikrein did not differ between patients with chronic abuse of analgesics and healthy volunteers. The excretion of kallikrein in patients with manifest analgesic nephropathy, however, was decreased. Thus the renal concentration test with DDAVP proved to be useful in early diagnosis of renal dysfunction caused by analgesics. PMID- 2569307 TI - Identification of the molecular structure of the phenolic primary metabolite of dimetindene in animals and man. AB - The metabolic pathway of dimetindene (dimetindene maleate, Fenistil), an antiallergic drug commercially available for more than 20 years, has remained unknown. By means of HPLC, GC-MS and MS-MS, dimetindene was found to be hydroxylated on the indene moiety of the molecule both in vitro with hepatic microsomes of several species and in vivo in man. Cochromatographic analysis (HPLC and GC-MS) of the primary metabolites produced in vitro and in vivo demonstrated their similarities but revealed differences from the chemically synthesized 5-hydroxy-dimetindene. Using large volumes of in vitro incubations of rat microsomes with dimetindene and cofactors, 1.15 mg of pure primary metabolite were first produced and then extensively purified. The study of the NMR proton in one and two dimensions (COSY) clearly demonstrated that this metabolite is hydroxylated on the C6 of the indene moiety as compared to the C5 for the synthetic product. PMID- 2569308 TI - [Screening and detection of quazepam and its metabolites]. AB - The paper describes a sensitive screening procedure for 7-chloro-5-(2 fluorophenyl)-1,3-dihydro-1-(2,2,2-trifluoroethyl)-2H-1,4- benzodiazepine-2 thione (quazepam, Oniria, Quazium) and its major metabolites. PMID- 2569309 TI - [The effect of the beta 1 selective beta blocker, betaxolol, on metabolism in type II diabetics]. AB - The effects of the highly beta 1-selective beta-blocker betaxolol (Kerlone) on carbohydrate and lipid metabolism in type II diabetic subjects were compared in a cross over study with those of a diuretic (xipamide-triamterene combination, Neotri). Betaxolol significantly lowered blood pressure with a tendency to better efficacy than the diuretic. The betaxolol-induced decrease in heart rate could be a contributing factor in attenuating the progression of atherosclerosis. No significant alterations were observed in postprandial blood glucose, glycosylated haemoglobin and 24-h urinary glucose excretion. The lack of noticeable influence on the carbohydrate metabolism during 4-week therapy is presumably related to the high beta 1-selectivity of betaxolol. Considering that diabetic hypertensive patients are at high risk for atherosclerosis it appears also favourable that betaxolol did not increase triglyceride levels and even decreased total cholesterol. The protective effect of beta-Blockers has been well established in the secondary and primary prevention of cardiovascular diseases. Especially diabetic patients might be expected to benefit from this therapy, if given preferably as beta 1-selective blocker that impairs carbohydrate and lipid metabolism in lesser extent. PMID- 2569311 TI - Genetic differences in the effects of voluntary ethanol consumption on brain monoamine levels in inbred strains of mice, C57BL/6J, C3H/He and DBA/2Cr. AB - This study was carried out to determine whether concentrations of the brain monoamines noradrenaline (NA), dopamine (DA), and 5-hydroxytryptamine or serotonin (5-HT) are different in three inbred strains of mice (C57BL/6J, C3H/He and DBA/2Cr) known to differ in their preference to alcohol, when they were given a free choice of 10% (v/v) ethanol and tap water to drink for 4 weeks. Mice of these three strains showed mean ethanol intakes of 4.41, 1.76 and 0.77 g/kg/day respectively. Levels of the above brain monoamines did not change in the alcohol preferring C57BL/6J mice, but in those with less preference for alcohol, C3H/He and DBA/2Cr, there were significant increases in DA and 5-HT levels respectively during the 4-week experiment. These findings suggest that inbred strains of mice show genetic differences of susceptibility to ethanol and that the strains with a low preference for alcohol undergo neurochemical changes after exposure to 10% ethanol and water even by free choice. PMID- 2569310 TI - Apolipoprotein A-I gene polymorphisms: frequency in patients with coronary artery disease and healthy controls and association with serum apo A-I and HDL cholesterol concentration. AB - We have investigated the association between serum high density lipoprotein cholesterol (HDL-C) and apo A-I concentration and the PstI and XmnI restriction fragment length polymorphisms of the apolipoprotein AI-CIII-AIV multigene complex. Two groups of subjects were examined. The first comprised 174 unrelated male patients under 60 years of age with angiographic evidence of coronary artery disease (CAD). Of this group 34 were non-North European. The second group consisted of 104 unrelated healthy male North European subjects aged under 60 and free from demonstrable CAD, who attended a health screening clinic in London. For the PstI polymorphism, the frequency of the rarer P2 allele was 0.12 in both the North European and non-North European patients and this was higher than in the control group (P2 frequency 0.06, P less than 0.05). Healthy individuals with the genotype P1P2 had higher levels of apo A-I but similar levels of HDL-C compared to those with the genotype P1P1. However, CAD patients with the genotype P1P2 had lower serum levels of apo A-I and significantly lower serum levels of HDL-C compared to those with the genotype P1P1 (0.85 mmol/l vs. 1.0 mmol/l, P less than 0.05). The allele frequencies of the XmnI polymorphisms were not significantly different in the control group and the group of North European patients, although within the sample of non-North European patients, the frequency of the X2 allele was significantly higher than that found in the North European controls (0.26 vs. 0.09). Patients with the genotype X1X2 had a higher mean serum concentration of HDL-C and apo A-I compared with patients with the genotype X1X1 (1.14 and 0.93 mmol/l for HDL-C, P less than 0.05; 147 and 123 mg/dl for apo A-I, P less than 0.05). Associations between HDL-C and apo A-I levels and PstI and XmnI genotype were similar in patients taking and not taking beta-blockers. The data show that genetic variation in the apo AI-CIII-AIV gene cluster is associated with coronary artery disease although only weakly, and suggest that the mechanism of this association may operate through an effect in determining the serum concentration of apo A-I and HDL-cholesterol. PMID- 2569313 TI - [Prostatic type papillary tumor of the urethra with endocrine cells. Immunohistochemical study]. AB - A peculiar variety of prostatic neoplasm harbouring a constellation of endocrine cells is reported. The tumor had a papillary growth projecting into the lumen of the prostatic urethra. The histological appearances were somewhat unusual. One fragment showed a typical feature of a prostatic-type polyp. However, much of the tissue were covered by a dysplastic epithelium. Adenomatous transformation of prostate-type polyp or "endometrial" adenocarcinoma could be considered. PMID- 2569314 TI - Pharmacology of the memory process. AB - Rather than reviewing once again the published literature on drugs which are capable of modifying the memory process, the authors stress the methodological difficulties which impede the development of such products. At the moment, substances which act on neurotransmission systems, and in particular cholinergic and gabaergic agents, seem to be good candidates. The choice of the molecule to be studied in man depends on the neurochemical physiopathology of the memory disorder which one seeks to correct. For this reason, one tries to select an animal model of memory which is closest to the clinical situation pertaining to man on which the tested molecule might act favorably. PMID- 2569312 TI - An experimental study of fetal alcohol syndrome in the rat: biochemical modifications in brain and liver. AB - Maternal alcohol consumption produces various abnormalities in the offspring, termed fetal alcohol syndrome. We investigated various biochemical modifications occurring in the brain and the liver of pups born to alcohol-consuming rats. The parameters analysed were: superoxide dismutase, a protector against free radicals injury, enolase isoenzymes as markers of nerve cell maturation, glutamine synthetase involved in ammonia detoxification, alcohol and aldehyde deshydrogenases in order to evaluate the contribution of acetaldehyde teratogenicity and ATPase activities involved in ion and neurotransmitter transport. Activities of all these enzymes were decreased in the brain even when alcohol was withdrawn from the mother diet either during pregnancy or lactation. Activities were also decreased in the liver, except enolase and alcohol deshydrogenase activities, which were increased, suggesting possible adaptative events in the presence of alcohol. It seems likely that the multiple alterations observed in experimental fetal alcohol syndrome may be caused by free radicals following decreased superoxide dismutase activity in addition to the toxicity of alcohol and its metabolites. PMID- 2569315 TI - The search for a biochemistry of memory. AB - The search for a biochemistry of memory may be divided into two consecutive periods: the search for molecular memory coding in the brain (1960-1975), and the ongoing search for biochemical correlates of learning or memory. During the first period, three main methods were used: chemical analysis of macromolecules after learning, studying the effects on learning of compounds (antibiotics) impairing macromolecule synthesis, and attempting chemical transfer of information items acquired by the brain ("memory transfers"). None of the three methods succeeded in clearly demonstrating that memory is chemically encoded. Subsequent modern work has focused on the search for correlates between brain chemical mechanisms and memory processes: the role of protein synthesis in memorization, peptide modulation of learning, and the role of the main neurotransmitter systems (particularly acetylcholine, catecholamines, GABA and glutamic acid). The correlates, which are to be interpreted with caution, are based on both pharmacological results in animals and clinical data in man. PMID- 2569316 TI - Animal studies of brain acetylcholine and memory. AB - Memory loss is a common feature of aging in the human but not all memories are equally lost. Usually the loss is more severe for the memory of recent events and information rather than the memory of remote events and information. Associated with memory losses of this type is a reduction in levels of cortical acetylcholine and a loss of cholinergic cells of the nucleus basalis magnocellularis. In the rat it is possible to assess two types of memory analogous to recent and remote memories in humans. Reference memory would refer to information concerning the task that is invariant from trial to trial. Working memory would refer to information that changes from trial to trial. In support of the hypothesis that decreases in cholinergic function lead to a differential impairment of working memory, rats trained in this task and given scopolamine showed a specific increase in working memory errors. In more recent studies rats have been trained in a task and then given unilateral neurotoxic lesions of the NBM. These results raise the intriguing possibility that degeneration of the NBM and associated memory impairment seen in aging and in Alzheimer's disease may be related to a change in the ratio of these or related endogenous tryptophan metabolites. PMID- 2569317 TI - Noradrenergic-cholinergic interaction: its possible role in memory dysfunction associated with senile dementia. AB - Over the past 10 years, cholinergic deficits have been strongly correlated with memory dysfunction. On the other hand the lack of efficacy of direct or indirect cholinergic drugs has led to research in other neurotransmitter systems. Several authors have found miscellaneous results in NE and metabolites levels and in monoamine enzyme activities. These discrepancies have lead to discussions about the homogeneity of the disease. Animal lesion studies point out the importance of the septo-hippocampal cholinergic pathway and the interrelations between ACh and NE in the hippocampal formation. The author performed a series of experiments in rats in which hippocampal ACh activity was reduced (fornix section or destruction of cells in the medial septum) and the NE system modified by clonidine or neurotoxic lesions. The hypothesis was that a lesion of the cholinergic pathway leads to an enhancement of NE activity which inhibits spared ACh neurons. The alpha-2 agonist clonidine would interrupt this loop by decreasing of NE release. Results provide arguments for the author's theory. PMID- 2569318 TI - Different affinities of alpha 2-agonists for imidazoline and alpha 2-adrenergic receptors. AB - It has recently been shown that imidazoline alpha 2-adrenergic agonists, such as clonidine and UK 14,304, selectively bind to both alpha 2- and imidazoline receptors in basolateral membranes from rabbit renal proximal tubule. In order to define the relative affinity of three antihypertensive alpha 2-agonists for the two classes of receptors, we performed competition studies of imidazoline alpha 2 antagonist 3H-RX 781094 and nonimidazoline antagonist 3H-rauwolscine binding to basolateral membranes from rabbit proximal tubule. The order of potency for inhibition of radioligand binding to basolateral membranes was rilmenidine greater than clonidine greater than guanfacine and clonidine greater than guanfacine greater than rilmenidine for 3H-RX 781094 and 3H-rauwolscine binding, respectively. These data show that not only clonidine, but also rilmenidine and guanfacine, drugs usually used as specific alpha 2-agonists, bind to both alpha 2 and imidazoline receptors. The higher affinity of these molecules for one or the other class of receptors could explain their different capacity to induce hypotension and side effects. PMID- 2569319 TI - Atrial natriuretic factor: neuromodulator of the central nervous system regulation of blood pressure. AB - Extensive examination in the mammalian brain for the presence of atrial natriuretic factors (ANF) has revealed both peptide and receptors specifically distributed throughout the central nervous system. High concentrations of ANF have been found in several hypothalamic nuclei, septal areas, the anteroventral third ventricular area (AV3V), and the median eminence, whereas moderate concentrations have been detected in the circumventricular organs and several brainstem nuclei. The receptors for ANF have been found in moderate to high concentrations in the olfactory lobe, AV3V region, and several circumventricular organs (subfornical organ, organum vasculosum of the lamina terminalis) as well as the nucleus tractus solitarius, median eminence, and choroid plexus. These findings suggest a role for ANF in modulating fluid and electrolyte balance, and blood pressure in this compartment, analogous to the proposed actions of this peptide hormone in the periphery. To determine whether ANF might function as a neuromodulator of blood pressure, we administered ANF via fourth ventricular injection into the brain of hypertensive (SHR) and normotensive rats (WKY). Atrial natriuretic factor caused a moderate and significant decrease in mean arterial blood pressure in both strains. The action of ANF appeared to be mediated by activating the central alpha 2-adrenergic nervous system, probably through the release of catecholamines. Further, a dependence on the secretion and action of an endogenous opioid was probably involved; heart rate was unaffected in these studies. Experiments from other laboratories indicate that central ANF may modulate the pressor effects of centrally acting angiotensin II.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569321 TI - A comparison of the efficacy of betaxolol and timolol in ocular hypertension with or without adrenaline. AB - Betaxolol hydrochloride, a selective beta 1-adrenergic blocker, is claimed to be less likely to aggravate chronic obstructive airways disease than timolol maleate, a nonselective beta-blocker. It is not clear which drug is more effective in lowering intraocular pressure (IOP) and how each acts in combination with topical adrenaline. To assess this we conducted a randomised clinical trial on ocular hypertensive patients. Both betaxolol and timolol produced a significant fall in IOP though not in all patients. No significant difference was found between betaxolol and timolol. The addition of dipivefrin gave an additional fall in some eyes only. No significant difference was found between the addition of dipivefrin and adrenaline. Some fellow eyes had a fall in IOP when dipivefrin was added to timolol. It is important to assess the response carefully in each individual eye when using betaxolol or timolol or when adding additional medication. PMID- 2569320 TI - Experimental compounds to lower intraocular pressure. AB - A need exists for new ocular hypotensive agents that are more efficacious and that have fewer side effects than those now clinically used. Laser-induced glaucoma in monkeys is an excellent model to test potential new drugs that lower intraocular pressure (IOP). A variety of agents that act through secondary messenger systems or via enzymes to lower IOP are being investigated in primates. Several alpha-adrenergic agonists and antagonists are effective ocular hypotensive agents in monkeys and humans. Para-aminoclonidine, an alpha2 agonist, inhibits the transient post-laser rise of IOP in humans. Prostaglandin F2 alpha-1 isopropyl ester significantly reduces IOP when tested in multiple dose fashion in glaucomatous monkey eyes and glaucoma patients. Modified carbonic anhydrase inhibitors, designed to enhance intraocular penetration, reduce IOP in the monkey model and in ocular hypertensive patients in single-dose studies when given topically. Studies show that forskolin and vanadate are less promising agents for glaucoma therapy. PMID- 2569323 TI - The benefits of prescription information leaflets (1). AB - 1. Prescription information leaflets (PILs) giving information about non steroidal anti-inflammatory drugs (NSAIDs), beta-adrenoceptor antagonists and inhaled bronchodilators were evaluated in three small Hampshire towns, while a fourth, in which no leaflets were distributed, acted as a control. 2. Seven hundred and nineteen (82%) patients prescribed one of these medicines agreed to be interviewed in their homes, 1 to 2 weeks after the medicine had been prescribed. Four hundred and nineteen of them had received leaflets, while 300 received no written information. Two hundred and sixty patients received their leaflets from a pharmacist while 159 were given them by their general practitioner. 3. Patients who received leaflets were better informed about every item of knowledge tested, except for the name of the medicine. Awareness of the side effects showed the greatest improvement, but there was no evidence that these leaflets produced spurious side effects. 4. Much improved levels of satisfaction were recorded amongst patients who received leaflets, especially those for NSAIDs (P less than 0.001) and for beta-adrenoceptor antagonists (P less than 0.01). 5. Subsequently, three hundred and fifty-eight (77%) of the patients prescribed either a NSAID or a beta-adrenoceptor antagonist 1 year earlier responded to a postal questionnaire. The benefits in terms of knowledge and satisfaction were still apparent, although less marked than previously. Of the patients still taking beta-adrenoceptor antagonists 70% had retained their leaflets over the intervening 12 months. 6. Ninety-seven per cent of patients read their leaflet regardless of whether it was distributed by a general practitioner or pharmacist. However, those who obtained it from a pharmacist tended to be more knowledgeable and satisfied. 7. We conclude that patients welcome the idea of receiving PILs. They improve patients' knowledge of how to take their medicines correctly and their awareness of potential side effects. Importantly, patients who receive leaflets are more satisfied than those who do not. These overall benefits justify the use of leaflets on a routine basis. PMID- 2569322 TI - Effect of CGP 17/582, a selective beta-adrenoceptor antagonist, on the haemodynamic and hypokalaemic response to adrenaline. AB - 1. CGP 17/582B is a new beta-adrenoceptor antagonist which on experimental studies appears to combine selective beta 1-adrenoceptor blockade with partial agonist activity (ISA). Assessing beta-adrenoceptor selectivity and the degree of partial agonist activity in vivo can be difficult. 2. In a double-blind placebo controlled crossover study we have compared the effect of oral pretreatment for 7 days with CGP (100 mg twice daily), with propranolol (non-selective beta adrenoceptor blocker with no ISA) and metoprolol (selective beta-adrenoceptor blocker with no ISA) on resting heart rate and heart rate response to submaximal exercise on a bicycle ergometer to assess the degree of beta-adrenoceptor blockade and also the changes in blood pressure, heart rate and potassium during the intravenous infusion of (-)-adrenaline to determine the degree of beta 2 adrenoceptor blockade. 3. Subjects underwent submaximal exercise testing on the second and fifth day of each treatment period and on the seventh day received a 2 h infusion of (-)-adrenaline (0.06 microgram kg-1 min-1). Heart rate, blood pressure, plasma potassium and catecholamines were measured throughout the study period. 4. All three active treatments significantly reduced exercise induced tachycardia. The (-)-adrenaline infusion significantly reduced plasma noradrenaline levels following propranolol and metoprolol and to a lesser extent with placebo but were unaltered on CGP. Baseline heart rate was unaltered by CGP but was significantly reduced by metoprolol and propranolol. Adrenaline significantly reduced plasma potassium levels following placebo and CGP pretreatment but plasma potassium was unaltered by adrenaline with metoprolol and propranolol pretreatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569324 TI - Pharmacodynamic and pharmacokinetic study of propranolol in patients with cirrhosis and portal hypertension. AB - 1. The aim of this study was to investigate the pharmacokinetics and the beta adrenoceptor blocking activity according to time of conventional (C) and long acting (LA) propranolol in cirrhotic patients. Twenty-four patients with alcoholic cirrhosis and oesophageal varices were randomly assigned to receive either 160 mg C propranolol, 160 mg LA propranolol or placebo acutely and then following repeated administration (acute and chronic phases). Thereafter propranolol concentrations and beta-adrenoceptor blockade (resting and exercise heart rates) were measured at different intervals. 2. The Cmax was significantly higher with C propranolol in both phases. The time of Cmax was significantly later with LA propranolol in both phases. The AUCs were significantly higher after chronic administration with both formulations of propranolol. 3. The exercise peaks of beta-adrenoceptor blockade were similar between the two formulations and between the two phases of administration of propranolol. The duration of effective beta-adrenoceptor blockade was significantly longer in the chronic phase and seemed to be longer with LA than with C propranolol although this was not significant (72 +/- 31 vs 48 +/- 18 h, respectively). 4. There was a significant correlation between the log propranolol concentration and exercise heart rate but not with resting heart rate. No correlation could be demonstrated between pharmacological data and the Child Pugh score. 5. We conclude that in cirrhotic patients exercise testing was a reliable method in the assessment of beta-adrenoceptor blockade. Pharmacology of propranolol was found to be different according to the formulation or to the phase of administration. PMID- 2569326 TI - Autologous blood stem cell transplantation in high-risk multiple myeloma. PMID- 2569325 TI - Effect of calcium antagonists on the chemosensitivity of two multidrug-resistant human tumour cell lines which do not overexpress P-glycoprotein. AB - We have examined the ability of eight compounds to enhance adriamycin (ADM) sensitivity of two human tumour cell lines (a small cell lung cancer cell line, NCI-H69, and a fibrosarcoma cell line, HT1080) and their multidrug-resistant variants. The resistant cell lines (H69AR and HT1080/DR4) do not overexpress P glycoprotein. Verapamil, nicardipine, perhexiline maleate, chloroquine, tamoxifen, clomiphene, prenylamine and trifluoperazine were tested alone and in combination with ADM for their cytotoxic effects. No major differences in sensitivity between the parent and resistant cell lines were noted when these agents were tested alone, except for HT1080/DR4 cells which exhibited a slight collateral sensitivity to nicardipine and H69AR cells which showed cross resistance to chloroquine and clomiphene. When the chemosensitisers were combined with ADM no enhanced cytotoxicity of either parent cell line was observed. In HT1080/DR4 cells, verapamil showed only a modest dose-dependent chemosensitising effect while the other compounds had no effect. Verapamil and nicardipine enhanced ADM cytotoxicity in H69AR cells slightly but these effects were not dose dependent. These results demonstrate that the reversal of drug resistance by verapamil and other calcium antagonists in a dose-dependent fashion is not an invariable property of multidrug-resistant tumour cells. PMID- 2569327 TI - Reconstitution and partial purification of the sodium and chloride-coupled glycine transporter from rat spinal cord. AB - The (Na+ + Cl-)-coupled glycine transporter has been solubilized from rat spinal cord with 2% cholate and purified 6-7-fold using Wheat Germ Agglutinin-Sepharose 4B. Transport activity - as determined upon reconstitution of the fraction into liposomes - was retained on the column and eluted by N-acetylglucosamine. When the glycoprotein fraction was depleted of the N-acetylglucosamine and applied to a second round of lectin-chromatography, the glycine transport activity was retained and again could be eluted by the sugar. The transporter activity reconstituted from the glycoprotein fraction retains the same features displayed in the synaptic plasma membrane vesicles, namely an absolute dependence on sodium and chloride, electrogenicity and efflux and exchange properties. These observations indicate that the (Na+ + Cl-)-coupled glycine transporter is a glycoprotein. PMID- 2569328 TI - [Inhibition of proliferation of normal and neoplastic liver cells under conditions of prolonged hormonal stimulation]. AB - Single injections of rats with hydrocortisone led to the inhibition of regenerating liver cell proliferation and protooncogene++ Ha-ras mRNA synthesis within 48 hours of hormonal induction. Administration of hydrocortisone to rats daily for 10 days resulted in a persistent decrease of the liver cell capacity to proliferate in response to partial hepatectomy. This inhibiting effect was observed for at least 7 days after cessation of hormonal stimulation; the level of Ha-ras mRNA was thereby decreased. A marked inhibition of ascite hepatoma cell growth was demonstrated after injections of those cells to mice induced with hydrocortisone for 10 days. Such a persistent effect of hydrocortisone is thought to be due to the depletion of the hormone-dependent hepatotrophic factors. The effect of the glucocorticoid hormone in vivo can be supposed to involve both the direct and indirect regulation of target cell proliferation. The latter is mediated via the changes in the activity of exogenous factors which control cell growth and proliferation. PMID- 2569329 TI - Activity of gamma-glutamyl transpeptidase in human fetal membranes. AB - The presence of gamma-glutamyl transpeptidase in human fetal membranes and alterations in enzyme activity during the gestational period were studied. Fetal membranes from term deliveries exhibited a high enzymatic activity, whereas membranes from preterm deliveries showed alterations with regard to the gestational weeks. These alterations were found to be similar to those which had previously been obtained from rat placentae during fetal growth and development. PMID- 2569330 TI - Changes of norepinephrine levels, tyrosine hydroxylase and dopamine-beta hydroxylase activities after castration and testosterone treatment in vas deferens of adult rats. AB - Norepinephrine levels and tyrosine hydroxylase and dopamine-beta-hydroxylase activities have been used to evaluate the effect of castration and testosterone treatment on the sympathetic innervation of the adult vas deferens. Castration was followed by a decrease in both norepinephrine content and tyrosine hydroxylase activity, even though the changes were not concomitant. Treatment of castrated animals with testosterone reversed the effect of castration on organ weight and norepinephrine content, but only a short-lasting increase in tyrosine hydroxylase activity occurred at the beginning of testosterone treatment. In contrast, the testosterone-induced recovery of norepinephrine content observed at this time was accompanied by a marked increase in dopamine-beta-hydroxylase activity. The results suggest that in rat vas deferens, norepinephrine levels are under androgenic control and that this regulation mainly involves changes in dopamine-beta-hydroxylase activity rather than a modulation of tyrosine hydroxylase. PMID- 2569331 TI - Formation of ovarian follicular cysts and corpora lutea after treatment with antihistamine or indomethacin in prepubertal gilts. AB - Roles of histamine and prostaglandins in the induction of ovarian cysts after unilateral ovarian manipulation (MAN) and in the process of ovulation were evaluated in prepubertal gilts treated with pregnant mare's serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). Administration of pyrilamine maleate, an H1 receptor antagonist, before MAN or hCG injection reduced the number of cysts formed but did not alter ovulation rate. Administration of cimetidine, an H2 receptor antagonist, failed to alter the incidence, number, or diameters of cysts formed in response to MAN or the ovulation rate on non-MAN ovaries. Administration of indomethacin, an inhibitor of prostaglandin synthesis, before MAN or hCG injection did not alter the incidence, number, or diameters of cysts formed on MAN ovaries but reduced the number of corpora lutea on non-MAN ovaries. Excessive accumulation of fluid in ovarian cysts apparently was mediated by histamine interacting with the H1-type receptor. Enhanced secretion of prostaglandins produced by the cyclooxygenase pathway did not contribute to development of ovarian cysts but, unlike histamine, was required for formation of corpora lutea in prepubertal gilts treated with PMSG and hCG. PMID- 2569333 TI - OKT4/OKT8 ratio in asymptomatic heroin addicts. AB - The OKT4 (helper) and OKT8 (suppressor) lymphocytic subpopulations were enumerated in a sample of 60 asymptomatic drug addicts and in 17 controls. No significant differences in the ratio could be found that could not be explained by the action of HIV. It can be concluded that heroin itself was not responsible for any alteration in the T4/T8 ratio in the population considered. PMID- 2569332 TI - Beta-adrenergic receptors in DMBA-induced rat mammary tumors: correlation with progesterone receptor and tumor growth. AB - In order to gain further knowledge about the potential role of catecholamines in mammary carcinoma, we have used the potent beta-adrenergic antagonist cyanopindolol (CYP) as iodinated ligand to characterize beta-adrenergic receptors in membranes prepared from mammary tumors induced by dimethylbenz(a)anthracene (DMBA) administration in the rat. The binding of [125I]CYP to membrane preparations of DMBA-induced rat mammary tumors is rapid at room temperature, reaching half maximal specific binding at 30 min of incubation. Scatchard analysis of the data indicates that [125I]CYP binds to a single class of high affinity sites (114 +/- 2.1 fmoles/mg protein) at an apparent KD value of 38.0 +/ 0.3 pM. The order of potency of a series of agonists to compete for [125I]CYP binding is consistent with interaction with a beta 2-subtype receptor: zinterol greater than (-)isoproterenol greater than (-)epinephrine much greater than ( )norepinephrine. In addition, the potency of a series of specific beta 1 and beta 2 synthetic compounds to displace [125I]CYP in mammary tumors is similar to their potency in typical beta 2-adrenergic tissues. The binding of [125I]CYP to DMBA induced rat mammary tumors shows a marked stereoselectivity, the (-)isomers of isoproterenol and propranolol being 150 and 80 times more potent, respectively, than their respective enantiomers. The autoradiographic localization of [125I]CYP performed on frozen sections revealed the presence of specific beta-adrenergic receptors in all the malignant cells. Spontaneous mammary tumors of aging (18-22 months) female rats have high levels of beta-adrenergic receptors. Castration decreased the concentration of [125I]CYP binding sites in DMBA-induced mammary tumors. A close correlation was observed between progressing, static, and regressing tumors after ovariectomy and beta-adrenergic receptor concentration. The presence of beta-adrenergic receptors in mammary tumors as well as the modulation of their level by ovarian hormones provides a mechanism for catecholaminergic influence in mammary cancer tissue. PMID- 2569334 TI - Value of radiographs in managing common foot injuries. PMID- 2569335 TI - Acute acquired methaemoglobinaemia after amyl nitrite poisoning. PMID- 2569336 TI - Beta blockers and carbohydrate and lipoprotein metabolism in hypertensive patients. PMID- 2569337 TI - Are beta blockers and nifedipine safe? PMID- 2569338 TI - Phaeochromocytomas and somatostatin production. PMID- 2569339 TI - A regional register for inherited cancers. PMID- 2569340 TI - Death after flumazepil. PMID- 2569341 TI - ABC of clinical genetics. Techniques of DNA analysis. PMID- 2569342 TI - Pressor effects of the alpha 2-adrenoceptor agonist B-HT 933 in anaesthetized and haemorrhagic rats: comparison with the haemodynamic effects of amidephrine. AB - 1. Blood pressure responses to single and multiple bolus doses of the alpha 2 adrenoceptor agonist B-HT 933 were analysed in intact anaesthetized rats which were either normotensive or hypotensive as a result of haemorrhage. Single bolus doses of B-HT 933 in normotensive rats induced a fall in blood pressure, whilst further doses induced dose-dependent pressor responses which were inhibited by the alpha 2-adrenoceptor antagonist yohimbine and unaffected by the alpha 1 adrenoceptor agonist prazosin. In the haemorrhagic, hypotensive animals, single bolus doses of B-HT 933 induced immediate dose-dependent pressor responses; the maximum pressor responses to the bolus of B-HT 933 and its ED50 values were the same in both the normotensive and hypotensive, haemorrhagic animals. 2. Cardiac output, its distribution and tissue blood flows were determined with tracer microspheres in intact anaesthetized normotensive and haemorrhagic, hypotensive rats during depressor (normotensive) and pressor (normotensive and hypotensive) responses to B-HT 933. Haemodynamics were also determined during pressor responses to the alpha 1-adrenoceptor agonist amidephrine. 3. In control normotensive rats, a single dose of B-HT 933 (1 mg kg-1) reduced blood pressure by reducing cardiac output (through a decrease in heart rate). It increased the fractional distribution of cardiac output to the spleen and stomach, reduced it to the heart and liver and reduced cardiac and hepatic blood flow. A further dose of B-HT 933 (1 mg kg-1 bolus followed by 100 micrograms min-1 infusion) increased blood pressure by increasing total peripheral resistance, which was accompanied by decreased proportions of cardiac output passing to the heart, liver and testes. There was also increased fractional distribution of cardiac output to the lungs, spleen, kidneys and stomach but blood flows through the liver and testes were reduced. Amidephrine (6 micrograms kg-1 bolus followed by 0.5 micrograms min 1 infusion) increased blood pressure by increasing cardiac output through an increased stroke volume. It increased cardiac output distribution to the kidneys and brain, increasing blood flow through the heart, lungs, brain, testes, epididimides, skin and large intestine. 4. Haemorrhage caused a fall in blood pressure which resulted from decreased total peripheral resistance and cardiac output (the latter due to decreases in both heart rate and stroke volume). It reduced the proportion of cardiac output distributed to the lungs, spleen, kidneys, testes and pancreas/mesentery and decreased blood flow through these organs as well as through the heart, liver, brain, epididimides, skin and the gastrointestinal tract.4 PMID- 2569343 TI - Prevention of physostigmine-, DFP-, and diazinon-induced acute toxicity by monoethylcholine and N-aminodeanol. AB - 1. Choline, and the choline analogues monoethylcholine (MEC) and N-aminodeanol (NAD) were examined for prophylactic activity in acute acetylcholinesterase inhibitor toxicity in mice. The rank order of potency of the compounds was MEC greater than NAD greater than choline. 2. Simultaneous administration of MEC (60 mg kg-1) or NAD (200 mg kg-1) with physostigmine reduced lethality to 17 and 13% respectively. MEC (60 mg kg-1) completely protected against disopropylfluorophosphate (DFP) and diazinon toxicity, and NAD reduced lethality to 17% for both agents. Choline (200 mg kg-1) exhibited only negligible antidotal activity against the inhibitors. 3. In vitro concentrations of choline, MEC, and NAD, similar to the estimated concentration obtained in vivo in the acute toxicity study, produced mixed inhibition of mouse brain acetylcholinesterase. The inhibition was dose-related and was additive to the inhibition produced by the cholinesterase inhibitors. 4. All three analogues reduced ligand binding at the nicotinic, M1, and M2 receptors. The rank order of potencies for the analogues at each receptor was nicotinic: (choline greater than MEC greater than NAD), M1: (MEC greater than choline greater than NAD), and M2: (MEC greater than choline greater than NAD). 5. It is proposed that the analogues prevent acetylcholinesterase inhibitor toxicity peripherally by interacting with acetylcholinesterase, and/or by competing with acetylcholine for binding to cholinoceptors. PMID- 2569345 TI - Subclassification of alpha 1-adrenoceptor recognition sites by urapidil derivatives and other selective antagonists. AB - 1. The affinities of urapidil derivatives and other antagonists for alpha 1 adrenoceptors labelled by [3H]-prazosin were determined on membranes of six different rat tissues. 2. Urapidil and its 5-acetyl-, 5-formyl- and 5-methyl derivative displaced [3H]-prazosin from alpha 1-adrenoceptor binding sites in a concentration-dependent manner which varied with tissue. IC50 values were lower in vas deferens, hippocampus and cerebral cortex than in heart, liver and spleen. For 5-methyl-urapidil, binding to two distinct sites could be demonstrated with mean K1 values of about 0.6 and 45 nM. Saturation binding studies with [3H] prazosin in the presence of 5-methyl-urapidil indicated a competitive type of interaction between 5-methyl-urapidil and [3H]-prazosin. 3. The proportion of [3H]-prazosin binding sites with high affinity for 5-methyl-urapidil was 58% in vas deferens, 69% in hippocampus, 41% in cerebral cortex and 23% in myocardium. In liver and spleen virtually no high affinity sites were found. These values were in good agreement with the percentages of binding sites with high affinities for WB-4101 and phentolamine, indicating that all these antagonists bind to the same subtype of alpha 1-recognition sites, whereas other alpha-antagonists like BE 2254, yohimbine and unlabelled prazosin did not discriminate between two binding sites. 4. Preincubating membranes of the cerebral cortex with chloroethylclonidine preferentially inactivated [3H]-prazosin binding sites with low affinity for 5-methyl-urapidil. 5. The antagonist potencies of 5-methyl urapidil and WB-4101 against alpha 1- adrenoceptor-mediated contractile responses were higher in vas deferens than in myocardium. The alpha 1-mediated effects in vas deferens but not in the heart were highly susceptible to nitrendipine. 6. Using 5-methyl-urapidil, the existence of two distinct alpha 1-adrenoceptor recognition sites could be demonstrated which correspond to the proposed alpha 1A and alpha 1B-subtypes. Since 5-methyl-urapidil is one of the ligands with most selectivity between these subtypes in binding studies it may serve as a valuable tool for such investigations. PMID- 2569344 TI - Beneficial effect of amosulalol and phentolamine on post-hypoxic recovery of contractile force and energy metabolism in rabbit hearts. AB - 1. The effects of phentolamine, an alpha-adrenoceptor blocking agent and amousulalol, an alpha 1 and beta-adrenoceptor antagonist on hypoxia-induced impairment in cardiac function and metabolism were examined using the isolated heart Langendorff preparation of the rabbit. 2. Hypoxia induced cessation of cardiac contractile force, a rise in resting tension, a decrease in myocardial high-energy phosphates, an increase in tissue calcium content and the release of ATP metabolites from the heart. Subsequent reoxygenation resulted in little recovery of cardiac contractile force, and there were further increases in tissue calcium content and in the release of creatine kinase from the heart. 3. Treatment of hypoxic hearts with either 83 microM phentolamine or 45 microM amosulalol resulted in a suppression of the rise in resting tension, the tissue calcium accumulation and the release of creatine kinase and ATP metabolites during hypoxia. This treatment also elicited significant recovery of cardiac contractile force, restoration of myocardial high-energy phosphates, suppression of the release of creatine kinase and the accumulation of tissue calcium during reoxygenation. Both 83 microM phentolamine and 45 microM amosulalol a significant prolongation of the effective refractory period of rabbit isolated atria. 4. Lower concentrations of phentolamine (16 microM) and amosulalol) (9 microM), which are sufficient to exert an alpha-adrenoceptor blocking action, did not elicit an appreciable effect on the post-hypoxic recovery of cardiac contractile force. 5. These results suggest that phentolamine and amosulalol are capable of protecting the myocardium from hypoxia-induced derangements in cardiac function and metabolism. This effect is probably attributable to their membrane stabilizing effect, rather than to their alpha-adrenoceptor blocking action. PMID- 2569346 TI - Mode of antinociceptive action of flupirtine in the rat. AB - 1. Flupirtine is a novel, centrally acting, non-opioid analgesic agent. The present investigation was undertaken to ascertain which neuronal systems might be responsible for its antinociceptive effect in rodents. The antinociceptive responses to the test compounds were examined in the tail-flick test. 2. The selective destruction of noradrenergic pathways by 6-hydroxydopamine considerably reduced the flupirtine-induced inhibition of nociceptive responses but not the clonidine-induced antinociception which was significantly enhanced. Depletion of spinal 5-hydroxytryptaminergic pathways by pretreatment with 5,7 dihydroxytryptamine failed to affect the action of flupirtine and clonidine. 3. The depletion of neurotransmitters by reserpine totally abolished the antinociceptive action of flupirtine. By contrast, clonidine-induced inhibition of nociceptive responses remained unchanged. 4. Inhibition of the synthesis of noradrenaline by alpha-methyl-L-p-tyrosine attenuated the antinociception induced by flupirtine. In contrast, inhibition of the synthesis of 5-hydroxytryptamine by (+/-)-6-fluorotryptophan did not influence the antinociceptive activity of flupirtine. 5. Inhibition of noradrenaline uptake by imipramine led to a significant augmentation of flupirtine-induced antinociception. 6. Selective antagonists at alpha-adrenoceptors significantly decreased the antinociceptive action of flupirtine. Antinociception induced by clonidine was significantly diminished by idazoxan but not by prazosin. 7. The 5-hydroxytryptamine (5-HT) antagonist, ketanserin diminished the antinociceptive activity of flupirtine, probably due to its additional alpha 1-adrenoceptor antagonist activity. The antinociceptive effect of clonidine was not influenced by ketanserin. 8. Cholinoceptor antagonists such as mecamylamine and pirenzepine did not alter the antinociceptive action of flupirtine. Flupirtine-induced antinociception also remained unchanged after pretreatment with haloperidol. 9. Flupirtine has no pharmacologically relevant affinity for alpha 1-, alpha 2-adrenoceptors, 5-HT1- and 5-HT2-receptors as shown in direct binding studies. 10. The present results indicate that the antinociceptive action induced by flupirtine depends on the descending noradrenergic pain-modulating system. PMID- 2569347 TI - Physical dependence on diazepam in the dog: precipitation of different abstinence syndromes by the benzodiazepine receptor antagonists Ro 15-1788 and ZK 93426. AB - 1. The effects of the benzodiazepine receptor antagonists Ro 15-1788 (flumazenil) and the beta-carboline ZK 93426 were compared in dogs before and after chronic treatment with diazepam. 2. In diazepam-naive dogs, the most prominent behavioural alterations occurring during or after i.v. infusion of Ro 15-1788 up to a dose of 20 mg kg-1 were transient sedation, ataxia, and 'hot foot' behaviour, whereas behavioural alterations observed after ZK 93426 were not different from those observed after i.v. infusion of vehicle alone. This indicates that, in contrast to Ro 15-1788, ZK 93426 did not exert partial agonistic activity at benzodiazepine receptors. 3. In dogs treated 3 times daily with diazepam, 1 mg kg -1 orally, for 1 week, both benzodiazepine antagonists precipitated abstinence symptoms but the number and severity of withdrawal signs induced by Ro 15-1788 were greater than with ZK 93426. 4. In dogs treated 3 times daily with diazepam, 2 mg kg-1 orally, for 2 weeks, severe abstinence symptoms were precipitated in all animals by infusion of either antagonist but differences were found in the type of the symptoms: Ro 15-1788 induced rigid postures or rigid walking with increased muscle tone, tremor, twitches and jerks, whereas ZK 93426 did not alter motility but induced generalized myoclonic jerks and tonic clonic seizures. A generalized tonic-clonic seizure was also observed in one dog of the trial with infusion of Ro 15-1788. 5. Plasma level determinations during chronic treatment diazepam showed marked accumulation of the major active metabolite desmethyldiazepam, whereas diazepam levels were at least 15 times lower, which might suggest that desmethyldiazepam was responsible for the development of physical dependence on diazepam. PMID- 2569348 TI - Investigation of the central sites at which morphine acts to cause hypertension in conscious rabbits. AB - 1. In conscious rabbits intracerebroventricular (i.c.v.) morphine (10 and 50 micrograms kg-1) caused a dose-related increase in plasma noradrenaline and adrenaline, respiratory depression and sedation. The increase in sympatho-adrenal outflow resulted in hypertension accompanied by bradycardia and the increase in adrenaline secretion caused hyperglycaemia. Morphine (1 microgram kg-1 i.c.v.) and i.c.v. saline had no effect. 2. The same doses of morphine given intracisternally (i.c.) caused bradycardia and a similar degree of respiratory depression to i.c.v. morphine, but no significant increase in blood pressure and only a small, gradual rise in plasma adrenaline. 3. Intravenous naloxone (1 mg kg 1) did not block the hypertension, hyperglycaemia or increase in plasma catecholamines that followed i.c.v. morphine, but prevented the respiratory depression and sedation. 4. Ganglionic blockade with pentolinium prevented the rise in plasma catecholamines, blood pressure and plasma glucose induced by i.c.v. morphine. 5. These findings demonstrate that the increased sympathoadrenal outflow following i.c.v. morphine results from an action on periventricular structures. The resultant increase in plasma catecholamines, which is largely naloxone resistant, accounts for the hypertension and hyperglycaemia. The bradycardia is probably partly baroflex mediated and partly due to an increase in vagal tone as a result of stimulation of brainstem opioid receptors. The respiratory depression is probably due to an action of morphine on brainstem opioid receptors. PMID- 2569350 TI - Development and selective neurodegeneration in cell cultures from different hippocampal regions. AB - Previous studies have shown that pyramidal neurons in hippocampal regions CA1 and CA3 are selectively vulnerable in several neurodegenerative disorders and that a subpopulation of pyramidal neurons in cell cultures of embryonic hippocampus are sensitive to glutamate neurotoxicity. In order to determine whether the patterns of cell loss seen in situ correlate with intrinsic differences in neuronal sensitivities to glutamate-induced degeneration acquired during development, we characterized cultures established from different regions of postnatal rat hippocampus and then examined neuronal sensitivity to glutamate. Tissue corresponding to the dentate gyrus (DG) and regions CA1, CA2 and CA3 of Ammon's horn was removed by microdissection from transverse hippocampal slices and was used to establish cultures of dissociated cells. Cultures from all 4 regions contained 3 major morphological classes of neurons; pyramidal-like, bipolar and stellate. Pyramidal-like neurons comprised the majority of neurons in all cultures; these neurons extended one long and branching axon, and one or more short dendrites. Immunocytochemistry showed that all neurons possessed high levels of glutamate-like and gamma-aminobutyric acid (GABA)-like immunoreactivity when grown in isolation. In contrast, when bipolar and pyramidal neurons were cultured in contact with glial cells, glutamate and GABA immunoreactivity were selectively reduced in the bipolar and pyramidal cells, respectively, suggesting that cell interactions influence neurotransmitter phenotype. Subpopulations of hippocampal neurons from each hippocampal region were vulnerable to glutamate induced neurotoxicity. Bipolar and stellate cells were resistant to glutamate, while pyramidal-like neurons showed varying degrees of sensitivity to glutamate depending upon which region they were taken from. Experiments with specific glutamate receptor agonists and antagonists demonstrated that both non N-methyl-D aspartic acid (NMDA) receptors and NMDA receptors mediated glutamate-induced degeneration. There were clear differences in the vulnerability of the pyramidal like neuron populations in cultures from the different hippocampal regions. The rank order of the vulnerability of pyramidal-like neurons to glutamate-induced neurodegeneration between regions in culture was: DG less than CA2 less than CA3 less than CA1. This pattern of selective vulnerability in cell culture corresponds directly to the pattern of selective cell loss seen in situ in Alzheimer's disease, epilepsy, and stroke suggesting that intrinsic neuronal differences in glutamate sensitivity may be involved in these disorders. PMID- 2569349 TI - Subpopulations of somatostatin 28-immunoreactive neurons display different vulnerability in senile dementia of the Alzheimer type. AB - We tested whether the vulnerability of somatostatin (SST) neurons in senile dementia of the Alzheimer type (SDAT) depended upon their co-localization with neuropeptide Y (NPY). Density estimates of SST28- and NPY-immunoreactive neurons and percentage of double-labeled SST-NPY neurons were obtained in the cortex (areas 9 and 25) and the bed nucleus of stria terminalis (BST), in 6 SDAT and 5 control cases. Counts of senile plaques (SP) and neurofibrillary tangles (NFT) were done on thioflavin S stains. In both cortical areas, a decrease in the density of SST28-IR neurons was found in SDAT cases (-60% in area 25 and -80% in area 9), whereas density of NPY-IR neurons was unchanged. Accordingly, the proportion of single-labeled SST neurons decreased; this decrease was significantly correlated with SP (r = -0.89, P less than 0.001). We conclude that single SST-IR neurons, in cortical layers II-III, and V, are preferentially lost relative to co-localized SST-NPY neurons. In the BST, no significant reduction of SST-IR, NPY-IR neurons nor of the percentage of single labeled SST neurons was found, despite the presence of SP. Thus one subpopulation of SST neurons, defined by associated neurochemical characters (not co-localized with NPY nor with NADPH diaphorase) and by topography (cortical layers III and V) appears to be particularly vulnerable in SDAT. The potential importance of their position in neural circuitry is emphasized. PMID- 2569351 TI - Twenty-four hour variation of alpha 1-adrenergic receptors in the pineal gland of the male Syrian hamster. AB - Specific binding of [125I]iodo-[beta-(4-hydroxyphenyl)-ethylaminomethyl]tetralone ([ 125I]HEAT) was used to assess alpha 1-adrenergic receptors on pineal gland membranes of male Syrian hamsters (Mesocricetus auratus) housed under a 14:10 h light-dark cycle (lights on at 06.00 h). Saturation experiments with pooled pineal membrane preparations showed the presence of alpha 1-adrenergic receptor sites (dissociation constant Kd approx. 0.1 nM). Analysis of 4 time points indicated no significant change in Kd, but significant (P less than 0.01) changes of receptor density (Bmax) with a minimum recorded at night. Binding of a constant amount of [125I]HEAT (200 pM) to pineal membranes at 8 time points exhibited a circadian variation (P less than 0.001) of receptor density with lowest values around midnight and highest levels during daytime. PMID- 2569352 TI - Intrathecal dynorphin A administration causes pressor responses in rats associated with an increased resistance to spinal cord blood flow. AB - Hemodynamic responses (blood pressure, as well as cardiac output (CO), peripheral and CNS blood flow changes measured via radioactive microspheres) were analyzed in anesthetized rats 2 min following intrathecal (IT) administration (10 microliters) of either 5-ion control solution or 20 nmol of dynorphin A(1-13) into the lower thoracic space (T10-T12). Mean arterial pressure (MAP) significantly increased within 2 min following IT dynorphin A(1-13) due to rise in total peripheral resistance, whereas CO significantly declined. Two minutes post-IT-dynorphin A(1-13) administration spinal cord blood flow also significantly decreased for 2 cm anterior and 1 cm posterior from the tip of the spinal catheter, which reflected a significant elevation in tissue flow resistance of spinal cord vessels in spite of the reduction of CO. As well, tissue blood flow resistance was also increased at this time in the kidneys and adrenal glands. The results indicate that within 2 min after intrathecal dynorphin A(1-13) administration an acute increase in blood flow resistance of spinal cord vessels around the tip of the spinal catheter occurs, at a time when the animal is also hypertensive. It is suggested that the associated pressor response may, in part, be caused by dynorphin A evoking localized ischemia. PMID- 2569353 TI - Central alpha 1 adrenoceptor subtypes in narcolepsy-cataplexy: a disorder of REM sleep. AB - The present study suggests the specific involvement within the central nervous system of an alpha 1 adrenoceptor subtype in a behavior, the control of cataplexy, a pathological analogue of rapid eye movement (REM) sleep atonia. Experiments have shown that prazosin, an alpha 1 antagonist, dramatically aggravates canine narcolepsy-cataplexy through a central mechanism, and that [3H]prazosin binding sites are increased in the amygdala of narcoleptic dogs. However, the corresponding Scatchard plots were curvilinear and best fit was obtained with a two-site model, suggesting the existence of two [3H]prazosin binding sites. These two sites (high and low affinity [3H]prazosin binding sites) met the criteria for authentic receptors and were respectively very similar to the alpha 1a and alpha 1b (high and low affinity for WB4101, respectively) subtypes recently described in the rat and rabbit. Our results of in vivo pharmacology and in vitro [3H]prazosin binding in canine narcolepsy now clearly implicate the low affinity [3H]prazosin binding site (alpha 1b) in canine narcolepsy: (1) Prazosin, an alpha 1 antagonist with similar affinity for both subtypes, was much more potent in increasing cataplexy than WB4101, a compound with more affinity for the alpha 1a receptor. (2) Chlorethylclonidine and phenoxybenzamine, two irreversible blockers of the alpha 1 receptors with more affinity for the alpha 1b receptors, aggravate cataplexy for up to two weeks. (3) The alpha 1 receptor upregulation previously reported by our group in the amygdala of narcoleptic dogs was due to a selective increase in the low affinity [3H]prazosin binding sites. A role for noradrenaline in REM sleep regulation has been suspected for many years, but has never been clearly elucidated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569354 TI - Participation of cyclic adenosine monophosphate and beta-adrenergic receptors in the facilitatory effect of noradrenaline on the response of cultured cerebellar neurons to glutamate. AB - For the purpose of examining possible involvement of the cyclic adenosine monophosphate (cAMP) system and adrenergic receptors in the modulatory effect of noradrenaline (NA) on the glutamate-induced depolarizing response, the effects of dibutyryl cAMP (DBcAMP), forskolin, theophylline, clonidine, isoproterenol and propranolol were intracellularly investigated in the cerebellar neurons cultured from chick embryos. Not only NA-induced hyperpolarization and increase in input resistance but also the facilitatory effect of NA on the glutamate response were mimicked by DBcAMP and isoproterenol. This facilitatory effect of DBcAMP was enhanced by theophylline or forskolin, while that of isoproterenol was antagonized by propranolol. Clonidine suppressed glutamate-induced depolarization. These results that the enhancing action of NA on the responsiveness of cultured cerebellar neurons to excitatory amino acids is mediated by beta-adrenergic receptors and the intracellular cAMP system. PMID- 2569355 TI - [Neuroendocrinology in psychiatry]. AB - The ideology of neuroendocrinological research in psychiatry as well as the most important findings in psychoneuroendocrinology are briefly reviewed. The paper is intended for postgraduate education in psychiatry and endocrinology. PMID- 2569356 TI - The effect of histamine-H1 receptor antagonism with terfenadine on concentration related AMP-induced bronchoconstriction in asthma. AB - Selective histamine-H1 receptor antagonists inhibit adenosine 5'-monophosphate (AMP)-induced bronchoconstriction by greater than 80% when expressed as a percentage inhibition of the FEV1 time-response curve following inhalation of the provocation concentration of AMP required to produce a 20% decrease in FEV1 from baseline (PC20). To investigate this further we have determined that, in eight mild atopic asthmatic subjects, terfenadine (180 mg), administered 3 hr pre challenge, increases the geometric mean PC20 for histamine from 0.4 (range 0.03 3) mg/ml after placebo, to 20.2 (range 0.6-64) mg/ml following active treatment (P less than 0.0001). For AMP, the PC20 increased from 9.3 (range 1.0-113.3) mg/ml after placebo, to 150.2 (range 32.1-1177.7) mg/ml with terfenadine (P less than 0.0001). This 16.2-fold (range, 5.5-47.9) displacement to the right of the AMP concentration-response curve by a selective histamine-H1 receptor antagonist emphasizes the central role of histamine in the airways response to this nucleotide. PMID- 2569357 TI - Intrathoracic ganglionic beta-adrenergic receptors involved in efferent sympathetic regulation of the canine heart. AB - The enhancement of cardiac inotropism induced by electrical stimulation of acutely decentralized efferent preganglionic sympathetic axons was reduced, but still present, following intravenous hexamethonium administration (10 mg/kg). This residual enhancement was eliminated following subsequent administration of timolol (0.1 mg in 0.1 mL normal saline) into the ipsilateral stellate and middle cervical ganglia. Similar results were obtained when the order of administration of these agents was reversed. When the beta-adrenergic agonist isoproterenol (1, 2 or 5 micrograms in 1 microL of normal saline) was administered into specific loci of an acutely decentralized stellate or middle cervical ganglion, cardiac chronotropism and/or inotropism were increased; when similar injections were made into adjacent ganglionic sites cardiac responses usually were not elicited. Following hexamethonium administration similar results were obtained indicating that these effects were not primarily dependent upon nicotinic cholinergic synapses. In contrast, repeat injections of isoproterenol into the site that initiated cardiac responses when injected with isoproterenol failed to elicit any cardiac responses following local administration of timolol into the same site indicating that the effects of isoproterenol could be blocked by a beta adrenergic receptor blocking agent. Injections of 1 to 5 micrograms of isoproterenol into a ganglion which was surgically disconnected from the heart failed to elicit cardiac responses, demonstrating that at these doses detectable activation of cardiac myocyte beta-adrenergic receptors via circulating isoproterenol which had leaked out of the ganglion did not occur. It is concluded that beta-adrenergic receptors in intrathoracic ganglia are involved in the efferent sympathetic regulation of the heart and that such receptors are associated with neurons in specific ganglionic loci. PMID- 2569358 TI - Chromosome 12q heterozygosity is retained in i(12p)-positive testicular germ cell tumor cells. AB - Restriction fragment length polymorphism analysis is used to demonstrate that formation of the i(12p) chromosome, characteristic of testicular germ cell tumors, does not lead to loss of heterozygosity of various loci on the q arm of chromosome 12. This result suggests that during the etiology of these tumors, aneuploidization precedes the formation of the i(12p) marker chromosome. PMID- 2569359 TI - Elimination of chemoresistant multiple myeloma clonogenic colony-forming cells by combined treatment with a plasma cell-reactive monoclonal antibody and a P glycoprotein-reactive monoclonal antibody. AB - Patients with multiple myeloma (MM) commonly become refractory to chemotherapy despite a favorable response to induction treatment. We examined the effectiveness of a previously characterized plasma cell-reactive monoclonal antibody, MM4, in eliminating MM clonogenic colony-forming cells (CCC) with a multidrug-resistant (MDR) phenotype. Experiments were performed using MM cell lines that exhibit 6 (RPMI 8226/DOX6)- and 40 (RPMI 8226/DOX40)-fold resistance to doxorubicin (DOX). Both lines were selected from the chemosensitive MM line RPMI 8226/S and were cross-resistant to mitoxantrone, acronycine, etoposide, and vincristine. Surface marker analysis conducted in this study showed that DOX6 and DOX40 overexpressed the MDR1 gene product p170. Both MDR lines remained reactive to the plasma cell-reactive monoclonal antibodies MM4 and PCA-1 and expressed the relevant cytoplasmic immunoglobulin light chain. Treatment with MM4 and rabbit complement (C') was equally cytotoxic to RPMI 8226/S [80 +/- 5.6% (SD)], DOX6 [74 +/- 8.5], and DOX40 cells [75 +/- 11.3%], based on short-term chromium release studies. Furthermore, MM4 + C' deleted up to 3 logs of CCC colonies from chemosensitive and MDR lines (RPMI 8226/S, 99.87 +/- 0.11%; DOX6, 99.91 +/- 0.08%; DOX40, 99.55 +/- 0.44%). By comparison, the P-glycoprotein-reactive monoclonal antibody MRK-16 and C' inhibited tumor colony formation of MDR cells (8226/DOX6, 95.71 +/- 2.51%; 8226/DOX40, 99.61 +/- 0.43%) but affected that of chemosensitive cells only slightly (8.9 +/- 17.8%). In an attempt to optimize the depletion of myeloma CCC, MM4 was used together with MRK-16. This approach resulted in uniform depletion of myeloma clonogenic colony-forming cells from the chemosensitive (98.32 +/- 1.53%, n = 4) and MDR lines (8226/DOX6, 98.83 +/- 0.08%, n = 4; 8226/DOX40 99.29 +/- 0.62, n = 7) but did not result in enhanced CCC depletion. When DOX40 cells were mixed with normal bone marrow (BM) in the ratio of 90:10 (BM:MM), either MM4 or MRK-16 and C' depleted MM colonies (98.8 +/ 0.71% and 98.10 +/- 1.0%, respectively) without affecting the majority of BM progenitor cells. These observations suggest that either MM4 or MRK-16 is useful for depleting MDR myeloma clonogenic colony-forming cells. PMID- 2569360 TI - [Pharmacologic characterization of the internal mammary artery used in myocardial revascularization]. AB - The aim of this study concerned the pharmacological investigation of the isolated internal mammary artery. Spirally-cut vascular segments were obtained from patients undergoing myocardial revascularization and set up in isolated baths under isometrical tension. Reactivity of internal mammary artery preparations to stimulatory and inhibitory agents was evaluated. KCl (90 mM), noradrenaline (10 8)-10(-5) M) and serotonine (10(-9)-10(-5) M) induced a tonic contraction lasting for more than 60 min. Angiotensin II (10(-6)-10(-5) M) and dopamine (10)-8)-10(4) M) resulted virtually uneffective. The serotonine-induced contractions were strongly inhibited by ketanserin (10-9)-10(-6) M), a selective S2-blocker, and also by verapamil (10(-3)-10(-6) M) and nitroglycerin (10(-7)-10(-5) M). These data suggest that internal mammary artery is sensitive to different contractile agents, in particular serotonine activates muscular contractions through S2 receptors. The knowledge of such a mechanism may be of clinical relevance. PMID- 2569361 TI - Regulatory pathways governing HIV-1 replication. PMID- 2569362 TI - A purified Drosophila homeodomain protein represses transcription in vitro. AB - even-skipped (eve) is a homeodomain-encoding gene that is a genetically defined repressor of Ultrabithorax (Ubx), fushi-tarazu (ftz), and wingless (wg). Here we report that purified eve protein represses transcription in vitro at the Ubx promoter, in a DNA binding site-dependent manner. eve protein represses transcription when bound either upstream or downstream of the RNA start site or when DNA binding sites are in either orientation. We also show that eve represses expression from the Ubx promoter in Drosophila tissue culture cells, again in a binding site-dependent manner. Deletion of eve DNA binding sites does not alter transcription in the absence of eve, and so repression is not likely to be the result of eve competitively inhibiting an activator protein from binding to the same DNA element. Instead, we propose that eve protein is probably interfering with the function of proteins bound at other locations in the promoter. The biochemical demonstration that a Drosophila homeodomain protein can directly regulate RNA synthesis strengthens the view that this class of regulators act as transcription factors to control development. PMID- 2569363 TI - The fission yeast cdc2/cdc13/suc1 protein kinase: regulation of catalytic activity and nuclear localization. AB - The products of the cdc13+ and cdc2+ genes form a stable complex that displays protein kinase activity in vitro. p63cdc13 is a substrate of p34cdc2, the catalytic subunit of the kinase. The histone H1 kinase activity of cdc2 oscillates during the cell cycle. Activation of the preformed cdc2/cdc13 complex at the G2/M transition requires cdc25+ gene function. Post-metaphase inactivation of the kinase is associated with loss of cdc13, which shares sequence homology with mitotic cyclins and, in common with these proteins, is degraded at each cell division. cdc13 and cdc2 co-localize in the cell nucleus. cdc2 is not degraded during mitosis, but in the absence of cdc13 it is not localized in the nucleus. These observations suggest that the cdc13+-encoded cyclin acts to regulate both the catalytic properties and the localization of the protein kinase of which it is a subunit. PMID- 2569365 TI - Atracurium and vecuronium interaction with gentamicin and tobramycin. AB - Drug interactions between the aminoglycosides (tobramycin and gentamicin) and atracurium and vecuronium were studied prospectively in 44 patients. Twenty-two patients had therapeutic serum levels of tobramycin or gentamicin and 22 served as controls. Onset time, clinical duration, and time to spontaneous recovery of T4/T1 ratio of 0.70 after atracurium or vecuronium injection were measured. No statistically significant differences were found in onset time, but clinical duration and time to recovery were significantly longer in patients receiving tobramycin or gentamicin and paralyzed with vecuronium than for controls (P less than 0.01 for clinical duration and P less than 0.0005 for recovery). The neuromuscular block produced by atracurium was not significantly influenced by the presence of therapeutic serum levels of tobramycin or gentamicin. We conclude that for patients treated with these antibiotics, atracurium may present some advantages over vecuronium when a prolonged block is not desired. PMID- 2569364 TI - A comparison of the incidence of pruritus following epidural opioid administration in the parturient. AB - Epidural morphine is associated with a high incidence of pruritus when used for pain control in the post-Caesarean section population. The purpose of this study was to compare the incidence of pruritus associated with epidural morphine, fentanyl, buprenorphine and butorphanol. Sixty healthy Caesarean section patients were studied in a double-blind randomized fashion. Patients were questioned at 1, 3, 12 and 24 hours postpartum for the incidence of pruritus. This study demonstrated that the incidence of pruritus was significantly higher following the use of epidural morphine and fentanyl. Even though epidural butorphanol and buprenorphine exhibited a low incidence of pruritus, their duration of analgesia was not long enough to make either attractive for single-dose administration. PMID- 2569366 TI - Histamine 2 receptor blocker in the treatment of protamine related anaphylactoid reactions: two case reports. AB - Two case reports are described of acute anaphylactoid reactions following the administration of protamine to reverse the anticoagulation effect of heparin in patients undergoing coronary artery bypass graft surgery. The administration of cimetidine seemed to reverse the anaphylactoid reaction after conventional treatment with epinephrine, H1 receptor blocker, and steroids had failed. We recommend that H2 receptor blockade be included with other drugs in the treatment of anaphylactoid reactions following protamine, and possibly after anaphylactoid reactions associated with other substances. PMID- 2569367 TI - Characterization of sulfasalazine's interference in the measurement of conjugated bilirubin by the Ektachem slide method. AB - We describe the cases of four patients who were taking sulfasalazine for inflammatory bowel disease, whose conjugated bilirubin concentrations in serum exceeded their corresponding total bilirubin concentrations as measured with a multilayer film analyzer, the Ektachem 400. Sulfasalazine added to pooled human serum at therapeutic concentrations increased the apparent conjugated bilirubin, as measured with the Ektachem, in a linear and dose-related fashion. Measured unconjugated bilirubin was simultaneously decreased to values less than -3 mg/L. The same interference occurred on the Ektachem 700, but an algorithm prevented the instrument from reporting the results. The major metabolites of sulfasalazine in blood did not interfere with analysis for those fractions of bilirubin. Sulfasalazine's strong absorbance at 400 nm explains its interference with determination of conjugated bilirubin in this instrument. PMID- 2569368 TI - Interaction between low density lipoprotein receptor (LDLR) and apolipoprotein E (apoE) alleles contributes to normal variation in lipid level. AB - Subjects drawn from a population-based register were studied with respect to lipid level association. The association of isoforms of apolipoprotein E (apoE) with lipid level in the general population was found to be limited to people with one particular genotype at the low density lipoprotein receptor (LDLR) locus. The results presented in this paper suggest that functional LDLR variants enhance or limit the effect of isoforms of apoE. The association between apoE4 and serum total and LDL cholesterol level may be mediated through the LDL (apoB100, apoE) receptor to a greater extent than previously thought. PMID- 2569369 TI - Non-allelic mutations in X-linked retinitis pigmentosa. AB - Using RFLP studies, the disease locus in two X-linked retinitis pigmentosa families was found to be centromeric to DXS7 in one family and telomeric to DXS7 in another, suggesting non-allelic heterogeneity. PMID- 2569370 TI - The goitre of the BB/O rat: an animal-model for studying the role of immunoglobulins stimulating growth of thyroid cells. AB - The BB rat is a well-known animal model for the study of autoimmune thyroid disease. Antithyroglobulin antibodies can be detected in the circulation from the age of 6 weeks onwards, and accumulations of lymphoid cells occurs in the thyroid of up to 60% of animals at the age of 20 weeks and over. The rat, however, stays euthyroid, the thyroid is not destroyed, and hence the disease is not yet well characterized. The study reported here shows that the thyroid weights of 41% (40/97) BB rats were raised from week 6 onwards in comparison to those of Wistar controls. Morphologically, BB thyroids showed a strong similarity to the human disease entity 'colloid goitre', namely an active growth, high columnar epithelium, branching and budding of thyrocytes, no signs of thyroid destruction and in 42% of rats at the age of 22-26 weeks, a development of intrathyroidal lymphoid tissue. Plasma TSH was not significantly raised in the animals. For this reason the presence of immunoglobulins which stimulate the growth of thyroid cells (so-called TGI's) was determined in 12-16-week-old BB rats (n = 10) and in control Wistar rats (n = 10). At this time a significant difference could be recorded in thyroid weights between BB/O rats and Wistar controls (20.1 +/- 6.0 mg vs. 15.8 +/- 2.9 mg, respectively), even in the absence of any intrathyroidal lymphoid cell infiltration. Protein-A-Sepharose purified serum IgG of these animals was used to detect TGI-activity via the 'Feulgen Cytochemical Bioassay'. Of the BB/O rats, eight were clearly positive for TGI, all of the Wistar rats were negative. The data show that the autoimmune prone BB rat may thus serve as an animal model for euthyroid goitre associated with thyroid stimulating antibodies. PMID- 2569371 TI - Immunoregulatory effects of alpha-endorphin, beta-endorphin, methionine enkephalin, and adrenocorticotropic hormone on anti-tetanus toxoid antibody synthesis by human lymphocytes. AB - This study assesses the effects of adrenocorticotropic hormone (ACTH), alpha endorphin (alpha-endo), beta-endorphin (beta-endo), and methionine-enkephalin (met-enk) on tetanus toxoid-stimulated in vitro specific antibody (anti-TT) synthesis. ACTH and TT-costimulated cultures enhanced antibody (Ab) synthesis over control TT-stimulated cultures at 10(-9) and 10(-11) M and suppressed Ab synthesis at 10(-13) and 10(-17) M. The alpha-endo and TT-costimulated cultures enhanced Ab synthesis over control cultures at all concentrations tested (10(-7) to 10(-15) M). Cultures costimulated with beta-endo and TT enhanced Ab synthesis over control cultures at 10(-11) M and suppressed Ab synthesis at doses above and below 10(-11) M. The met-enk and TT-costimulated cultures produced more Ab than controls at 10(-9) M; and the met-enk and TT-costimulated cultures produced less Ab than controls at other concentrations as low as 10(-17) M and as high as 10( 7) M. The results of this study provide one reason as to why the interactions between the neuroendocrine and immune systems remain unclear. PMID- 2569372 TI - Effects of the baroreceptor reflex system on atrial natriuretic factor secretion during volume expansion in dogs. AB - 1. The purpose of this study was to determine whether the baroreceptor reflex system affects the secretion of atrial natriuretic factor directly or indirectly during acute volume expansion. 2. Lactated Ringer's solution containing low mol. wt. dextran was infused at 20 ml/kg for 1 h into dogs in which baroreceptors had been denervated surgically (Vx), dogs in which the autonomic system had been blocked with hexamethonium (Hx) and control dogs. 3. The plasma noradrenaline level was significantly higher in the Vx group and lower in the Hx group than in the control group throughout the experiment. The plasma levels of arginine vasopressin in the Vx and Hx groups were significantly higher than in the control group. 4. The plasma atrial natriuretic factor levels in the three groups showed similar increases during and after volume expansion. 5. The plasma atrial natriuretic factor level was significantly correlated with the right atrial pressure during volume expansion. 6. From these results, it seems unlikely that changes in the plasma level of atrial natriuretic factor during volume expansion are regulated by the baroreceptor reflex directly or indirectly by systemic changes in the sympathetic nervous system or arginine vasopressin secretion. PMID- 2569373 TI - Permeability of the urinary bladder of the turtle, Pseudemys scripta elegans, to ammonia. AB - 1. The permeability of the isolated turtle urinary bladder to ammonia was investigated by varying the ratio of NH3 to NH4+ concentration in the serosal bath and measuring the flux of ammonia into the mucosal bath. 2. The permeability of the turtle bladder to NH3 was in the range of 5.4-6.1 x 10(-3) cm/sec and the permeability of the unstirred layer to NH4+ was in the range of 1.1-2.1 x 10(-5) cm/sec. 3. The flux of ammonia calculated from the permeability of NH3 can account for the decrease in mucosal H+ secretion observed with addition of NH4Cl to the serosal bath. PMID- 2569374 TI - Levels of T4, T3 and cortisol in the blood serum of the European bison (Bison bonasus) in the winter period. AB - 1. In the blood of 56 European bison, the levels of T4, T3, and the ratio of T3:T4 and cortisol were studied. 2. Between December and March, the T4 level changed from about 53 to 83 ng/ml of blood serum but in April it increased to 90 ng/ml. 3. The level of T3 in this period was 0.817-1.475 ng/ml and in April it was 2.40-3.40 ng/ml. 4. The ratio of T3:T4 was 0.015-0.024 and in April it increased to 0.027-0.052. 5. The level of cortisol changed from 1.00 to 6.70 ng/ml and in April it was from 1.00 to 14.00 ng/ml. PMID- 2569375 TI - Autoregulation of apical sodium entry in the colon of the frog (Rana esculenta). AB - 1. Na transport (INa) in the K-depolarized colon of the frog was investigated by electro-physiological current-voltage analysis. 2. INa and the intracellular Na activity [(Na)c] increased with increasing mucosal Na concentration ([Na]m), whereas the apical Na-permeability (PNam) and the transepithelial resistance (RT) decreased. 3. The results are consistent with a Na self-inhibition mechanism; however, a feedback inhibition of INa by intracellular Na must also be considered. PMID- 2569376 TI - Ammonia metabolism during the suckling period in the rat. AB - 1. The time-courses of blood glutamine, glutamate, alanine, ammonia, urea and allantoin concentrations during the first 15 days of extrauterine life were studied. 2. Glutamine and glutamate concentrations followed the same pattern and correlated positively, suggesting that both amino acids are utilized or released synchronously. 3. Alanine concentrations decreased during the first 3 days, reaching levels close to those of adults which persisted up to the end of the observation period. 4. A highly significant correlation was found between ammonia and urea concentrations, suggesting that during the suckling period urea synthesis may be limited by blood ammonia availability. 5. The time-course of allantoin concentrations suggests that the synthesis of purines was enhanced during the first day, decreasing sharply during the 2nd to steeply increase to the end of the suckling period. PMID- 2569377 TI - Fatty acid synthetase in control, starved and refed Richardson's ground squirrels. AB - 1. Starvation for 6 days reduced whole body mass and total body lipids to 76 and 71%, respectively, of pre-starvation levels in weight-gain phase ground squirrels. After 4 days of refeeding, body mass increased to 86% of pre starvation level but total body lipids had not changed from starvation levels. 2. Compared to the fed state, fatty acid synthetase (FAS) activity in white adipose tissue (WAT) was 15 and 31% in starved and refed 4-day animals, respectively, and in liver was 26 and 21% in starved and refed 4-day animals, respectively. Lipids depleted by starvation during prehibernatory fattening were not rapidly restored in Richardson's ground squirrels. 3. Changes in these parameters with starvation and refeeding were similar in weight-loss phase animals. 4. In control animals of both phases, WAT accounted for at least 90% of total FAS activity and liver nearly all of the remainder. PMID- 2569378 TI - Comparative aspects of microvillus development in avian and mammalian enterocytes. AB - 1. Pieces of chicken mid-jejunum have been used to determine microvillus length in enterocytes located at different points along the crypt-villus axis to determine the positional dependence of microvillus elongation. 2. The rate at which enterocytes migrate along the crypt-villus axis has also been measured, by injecting tritiated thymidine into the same animals, to determine the time course of microvillus elongation. 3. In mammals it had been shown previously that the maximal rate of microvillus elongation correlated closely with the physical size of the intestinal crypt (Smith et al., 1984). Comparison of crypt size with maximal rate of microvillus growth in chickens showed this correlation to apply to both animal classes. 4. The maximal microvillus length of a chicken enterocyte was much longer (3.3 microns) and the rate of enterocyte migration much less (4.0 microns/hr) than values reported previously for mammalian intestines. 5. The possibility that some other factor in addition to crypt size is affecting microvillus growth is discussed. PMID- 2569379 TI - Dual control over microvillus elongation during enterocyte development. AB - 1. Previously determined logistic growth constants describing enterocyte microvillus development for a variety of species were analysed for possible interactions taking place between enterocyte migration rate (R) and the size of individual crypts (CD). 2. Microvillus elongation, the c-value of a logistic growth curve, was found to increase linearly with crypt depth and reciprocally with decreasing migration rate. The starting microvillus length of a basal crypt enterocyte, the a-value, also increased linearly with CD without being affected by R. 3. The mathematical equation describing the effects of CD and R on M, the maximal microvillus length, was M = 0.0016 CD + 0.073 CD/R, where M and CD are measured in micron and R in micron/hr. 4. The relationship found between R, CD and M is explained by suggesting that the crypt environment enables enterocytes to respond to an initiating signal imposed on cells as they begin to migrate onto villi. The possible nature of this putative signal is also discussed. PMID- 2569380 TI - Carnosine content of the middle gluteal muscle in thoroughbred horses with relation to age, sex and training. AB - 1. Muscle biopsies were collected from 85 thoroughbred horses and analysed for carnosine content by an automated HPLC method. 2. No significant sex difference was found between colts, geldings and fillies. 3. There was a trend towards lower muscle carnosine contents with age, which was only significant between 1-year-old untrained horses and 4+ year-old horses (P less than 0.002). PMID- 2569381 TI - Ceftriaxone for the treatment of primary and secondary syphilis. AB - In a multicenter, open, randomized, prospective, comparative study, 28 patients with primary (n = 9) or secondary (n = 19) syphilis were treated with either ceftriaxone or penicillin G. 14 patients received ceftriaxone 4 x 1 g i.m. every 2 days. 14 other patients were treated with penicillin G 1 million IU i.m. daily for 15 days (standard therapy of primary and secondary syphilis). To avoid Herxheimer reaction we applied prednisolone 50-100 mg i.m. before specific treatment. Diagnosis was confirmed by clinical symptoms, dark-field microscopy and serological tests (VDRL titer, TPHA test and 19s-IgM-FTA-abs test or SPHA test). Follow-up examinations during therapy were repeated on days 2, 4, 6 (dark field microscopy, physical examination) and day 14 (VDRL titer). To evaluate therapeutic efficacy, serological controls were repeated 1, 2, 3, 6 and 12 months after therapy (VDRL, SPHA). In all patients, a marked decline (minimum 2-dilution decrease) in VDRL titer and resolution of clinical symptoms were noted. An adverse reaction was seen in 1 patient of the clemizol-penicillin G group (allergic penicillin exanthema). There were no adverse reactions to ceftriaxone. Summing up, there was no detectable difference in clinical and serological response to syphilis treatment either with ceftriaxone or penicillin G. Ceftriaxone, thus can be regarded as an equivalent alternative to penicillin G in the treatment of primary and secondary syphilis. PMID- 2569382 TI - Cloning of apoprotein AI gene and observation on DNA polymorphism of apoprotein AI/CIII gene-loci in genomic library from a liver of Chinese. AB - For the research on the structure and function of the HDL apoproteins, we have successfully constructed a complete genomic library, in self-prepared EMBL3 lambda vector and the packaging extracts of lysogenic strains BHB2688, BHB2690, from a Chinese fetal liver, which will be widely used in the research on the structure and function of apoproteins. In this study, a DNA sequence polymorphism, revealed by digestion of human DNA with the restriction endonuclease Sst1 and hybridization with an apoprotein AI cDNA probe, has been shown to be located at apoAI/CIII gene-loci. Three gene related fragments (5.7 Kb, 4.0 Kb and 3.0 Kb) were detected. The 5.7 Kb fragment is common and the polymorphism is demonstrated by the presence of either a 4.0 Kb fragment (S1 allele) or the 3.0 Kb fragment (S2 allele). Individuals were genotyped S1S1 or S1S2. Our result showed that the frequency of genotype S1S2 was higher in hypertriglyceridemic subjects than that in normolipidemic subjects. PMID- 2569383 TI - [Clinical significance of serum AKP, HSAP, LAP and gamma-GT determination in high risk pregnancy]. AB - Serum levels of alkaline phosphatase (AKP), heat stable alkaline phosphatase (HSAP), leucine aminopeptidase (LAP) and gamma-glutamyl transferase(gamma-GT) were determined in 249 blood samples from 202 women with normal or high-risk pregnancy at 30-42 weeks. The results showed that AKP and HSAP increased with the advance of gestational weeks (r = 0.9 259, P less than 0.01), reaching their plateau at 37-38th week. In severe hypertensive pregnant woman, AKP and gamma-GT were significantly elevated, while HSAP remained in consistent low level. A "dissociation phenomenon" between AKP and HSAP was discovered. In cases of prolonged pregnancy, oligohydramnios and IUGR with fetal distress and neonatal asphyxia, both AKP and HSAP were lower than those in normal pregnancies of same gestitional age. Serum level of HSAP might reflect the placental function, while combined with AKP and gamma-GT could also be helpful in assessing the severity of hypertension in pregnancy and in predictive feto-maternal prognosis. PMID- 2569384 TI - Analgesia in myocardial infarction. AB - The treatment of pain in the acute phase of a suspected acute myocardial infarction is often insufficient and has remained unchanged during recent years. The introduction of substances with a potential to limit the infarct size, such as thrombolysis and beta-blockade, have, however, decreased the requirement for narcotic analgesics (which are still the drugs of choice in many hospitals). Knowledge is still lacking regarding the duration of pain relief, the time between drug administration and pain relief, and optimal doses for various analgesics. Future research should aim at the development of drugs with a more rapid onset of action, less side effects and more complete analgesia. PMID- 2569386 TI - [Nonspecific effects of beta-adrenoblockaders]. PMID- 2569385 TI - [Immunomorphologic study of various endocrine cells in chronic gastritis and in intestinal metaplasia of the stomach]. AB - The amount of some endocrine cells in various forms of chronic gastritis with and without the presence of intestinal metaplasia of the stomach mucosa was studied immunohistochemically (PAP-reaction). The study was carried out on 18 samples of biopsied material for D-cells and on 31--for G cells. It was established that the amount of these cells as well as their ratio was changed in accordance with the form of chronic gastritis. Thus, for example++ , the number of D-cells was increased in mild forms of gastritis, but they were reduced in its grave form, while the number of G-cells gradually diminished from mild to grave forms. These changes give foundation to assume that hr hormonal disturbance of the stomach mucosa plays important role in pathogenesis of this disease. This suggestion is supported by the observed in dynamics quantitative changes of these cells, beginning even at the initial stages of this disease. PMID- 2569387 TI - [Interrelation of degradation processes of purine nucleotides and their biosynthetic transformation in the blood cells of patients with acute leukemia]. AB - It has been shown that the leukemic process shifts the balance of individual enzymatic steps of the purine nucleotide degradation and biosynthesis. In acute lymphoblastic leukosis (ALL) and acute myeloblastic leukosis (AML) the rate of purine degradation, as estimated by the ADA enzyme activity (ES 3.5.4.4.) and PNP (EC 2.4.2.1), exceeds considerably that of their enzymes AMP-DA (EC 3.5.4.6.) and IMP-DH (EC 1.2.1.14.) biosynthesis. The data obtained suggest the existence of differences in the enzymatic program of purine metabolism in the malignant transformation of hematopoietic tissue and in other tumours. PMID- 2569388 TI - [Endocrine changes in bulimia]. AB - There is a real organization of feed choice, with good equilibrium between neuropeptides and neurotransmitters actions, leading to choice and stimulation feeding. Morley described two main systems who control carbohydrate and lipids intake. These data contribute to future therapeutic research. PMID- 2569389 TI - [Chemotherapy in the treatment of bulimic behaviors]. AB - Bulimia has been treated with various drugs. MAOI and heterocyclic non MAOI antidepressants have proved efficacious in controlled studies. Whereas Benzodiazepines, Lithium and antiepileptic medications appear less promising or have not yet been widely assessed in methodologically valid studies. Serotoninergic compounds are particularly specific of bulimic disorders. The links of bulimia with depressive states are briefly discussed. PMID- 2569390 TI - Maternal modulation of growth hormone secretion in the neonatal rat. I. Involvement of milk factors. AB - The present experiments investigated the time course of maternal modulation of GH secretion and examined the possible role of milk in the regulation of GH secretion in neonatal rats. Serum GH concentrations in neonatal rats were high at birth and declined over time postpartum. Separation of rat pups from their mothers decreased, while a subsequent period of suckling increased, serum GH levels in rat pups on postpartum days 1-14, but not on day 20. The water-soluble fraction (infranatant) of rat milk contained immunoreactive (ir) rat GH-releasing hormone (rGHRH)-like material (725.06 +/- 81.29 pg/ml), ir-somatostatin-like activity (1.64 +/- 0.2 ng/ml), and irGH (4.79 +/- 0.73 ng/ml). The concentrations of these hormones tended to decrease with time postpartum and were positively correlated with each other (r = 0.70; P less than or equal to 0.0001). IrPRL was also present in the infranatant (148.44 +/- 14.55 ng/ml), but levels were not correlated with the other hormones detected. Milk infranatant stimulated GH secretion from perifused neonatal rat pituitary glands in vitro. Milk infranatant also stimulated GH secretion in vivo when administered sc or intragastrically to 2- or 8-day-old rat pups. The GH-releasing effect was not due to gastric distension or nonspecific nutritive components, as neither 0.9% saline nor nutrients (5% glucose and 10% BSA) increased serum GH levels. The presence of high concentrations of irGHRH in rat milk infranatant and the strong correlation between the irGHRH concentrations of milk samples and the in vitro GH response (r = 0.71; P less than or equal to 0.005) suggested that this peptide is a major candidate for producing the in vitro and in vivo GH-stimulating activity in rat milk. However, the minimally effective concentration of synthetic rGHRH required to stimulate GH release in the superfusion system was between 1-10 nM, which exceeds milk irGHRH levels by 100- to 1000-fold. Moreover, in vivo administration of synthetic rGHRH (sc or intragastrically) was unable to increase serum GH concentrations in 2-day-old pups, although a large dose (100 ng/g) of human GHRH sc was effective. These findings indicate that rat milk may be an important maternal factor that modulates GH secretion and, consequently, growth during the neonatal period. Rat milk has GH-releasing activity both in vivo and in vitro in neonatal rats, but the GH-releasing activities of milk are probably only minimally due to its rGHRH content. PMID- 2569391 TI - Effects of active immunization against somatostatin on serum growth hormone concentration in growing pigs: influence of fasting and repetitive somatocrinin injections. AB - Three experiments were conducted with growing pigs actively immunized against a protein-conjugated somatostatin (SRIF) in Freund's adjuvant. In the first experiment, blood from 24-week-old pigs (seven immunized and eight control) was sampled at 20-min intervals for 6 h to evaluate basal GH concentrations. The animals were then injected iv with porcine GH-releasing factor (GRF)-(1-29)NH2 (10 micrograms/kg). Before GRF stimulation, immunized animals had higher (P less than 0.05) baseline mean GH levels (2.6 vs. 1.4 ng/ml) and area under the GH curve (AUC; 1632 vs. 779 ng/min.ml); they also had higher AUC after GRF administration (4268 vs. 1972 ng/min.ml). In a second experiment eight immunized and eight control pigs were injected iv four times at 90-min intervals with porcine GRF (10 micrograms/kg). Control pigs responding to the first injection did not respond to the second and third, and those responding to the second did not respond to the first, third, and fourth, indicating a decreased responsiveness that was longer than 3 h post-GRF response in control pigs. SRIF immunized pigs had a more consistent GH response to the GRF injections. Overall, a reduced response was observed after the second and the fourth injections in immunized pigs, although five and six of eight animals had a GH peak response higher than 10 ng/ml during these periods. In a third experiment, effects of fasting, GRF, and SRIF immunization were studied. Immunization and fasting had their own positive effects on serum GH levels. Immunization increased baseline mean GH levels (5.0 vs. 2.2 ng/ml) and total AUC before (2318 vs. 1073 ng/min.ml) and after (1886 vs. 910 ng/min.ml) iv GRF stimulation (10 micrograms/kg) compared to controls. Fasting increased the mean baseline GH level (4.5 vs. 2.6 ng/ml), and it increased AUC before exogenous GRF stimulation (2009 vs. 1392 ng/min.ml). In conclusion, SRIF in pigs seems to be a potent GH-governing factor, since, when inhibited, baseline mean GH levels increase, and a consistent response to GRF is observed. Fasting could increase GH concentrations by different ways: decreasing SRIF release and increasing GRF release or modifying the sensitivity of the somatotrophs to both factors. PMID- 2569392 TI - A dominant phenocopy of hypopituitarism in transgenic mice resulting from central nervous system synthesis of human growth hormone. AB - We have produced a line of transgenic mice in which expression of human GH has been detected only in the cerebral cortex. Both male and female transgenic mice are growth inhibited with respect to their nontransgenic littermates. Mouse GH mRNA and insulin-like growth factor-I mRNA levels in the pituitary and liver, respectively, are reduced, and circulating insulin-like growth factor-I levels are lower in these mice. Within the hypothalamus somatostatin mRNA levels are increased and GH-releasing factor mRNA levels are reduced compared to those in nontransgenic littermates. We suggest that the growth retardation in these mice is a consequence of the ectopic human GH disturbing the normal controls that regulate mouse GH synthesis and release from the pituitary. These mice provide a resource for analysis of the regulation of GH production and demonstrate that a dominant phenocopy can be made by producing transgenic mice that have local production of an extra-cellular hormone. PMID- 2569393 TI - Central nervous system-mediated glucagon secretion is enhanced by alpha 2 adrenoreceptor activation. AB - We assessed the response of the adrenergic receptor in pancreatic glucagon secretion to central nervous system stimulation. Injection of neostigmine (5 x 10(-8) mol) into the third cerebral ventricle in intact rats resulted in increased epinephrine and norepinephrine secretion associated with glucagon secretion. This glucagon secretion was still observed in bilateral adrenalectomized (ADX) rats, although its concentration was significantly lower than that in the intact rats. This glucagon rise was significantly inhibited by ip treatment of ganglionic blocker with hexamethonium. Intraperitoneal injection of alpha-adrenergic receptor antagonist phentolamine (5 x 10(-7) mol), but not of beta-adrenergic receptor antagonist propranolol (1 x 10(-6) mol), reduced the hyperglucagonemic effect of a subsequent neostigmine injection in intact and ADX rats, although these antagonists did not influence epinephrine or norepinephrine secretion in intact rats. In addition, ip injection of the selective alpha 2 receptor antagonist yohimbine (5 x 10(-7) mol), but not of the selective alpha 1 receptor antagonist prazosin (1 x 10(-6) mol), inhibited the neostigmine-induced glucagon secretion in intact and ADX rats. From this evidence it is suggested that central nervous system-mediated glucagon release is enhanced by alpha 2 adrenoreceptor stimulation by either catecholamines or the autonomic nervous system. PMID- 2569394 TI - Spread of analgesia and ventilatory response to carbon dioxide following epidural somatostatin. AB - The effects of somatostatin, injected into the epidural space, on analgesia and control of ventilation were studied in 25 patients aged 41 +/- 9 yrs (mean +/- SD). The patients were allocated to three groups to receive: Group I--1 mg of somatostatin in 2 ml saline (n = 13); Group II--1 mg of somatostatin in 10 ml saline (n = 6); and Group III--somatostatin in a loading dose of 250 micrograms followed by an infusion of 125 micrograms h-1 (n = 6). Segmental cutaneous analgesia, assessed by pinprick, without loss of thermal sensibility or motor blockade was found in all patients. Onset times and durations of analgesia were 15 +/- 2 min and 69 +/- 19 min (mean +/- SD) in Group I and 14 +/- 2 min and 68 +/- 11 min in Group II. The extent of dermatome analgesia at 30 min and 60 min after somatostatin injection, respectively, was: T6 +/- 2 to T12 +/- 1, T4 +/- 2 to L1 +/- 2 in Group I, and T7 +/- 3 to L1 +/- 3, T3 +/- 1 to T12 +/- 2 in Group II. Continuous analgesia with onset of 16 +/- 2 min and extending from T7 +/- 1 to T12 +/- 1 was observed in Group III. No side-effects were observed. The control of ventilation studies in eight patients in Group I by the Read's rebreathing method did not show any significant change. PMID- 2569395 TI - Evaluation of the use of S1 nuclease to detect small length variations in genomic DNA. AB - A method which utilises S1 nuclease to detect small length variations in cloned and genomic DNA has been evaluated. The methodology of this technique is simple and robust, permitting the rapid analysis of 10(4) base pairs. By employing defined sequence variants, this method is shown to have a sensitivity which should enable the detection of length variations of only a few base pairs in heterozygous individuals. PMID- 2569396 TI - Protein kinase C stimulates adenylate cyclase activity in prolactin-secreting rat adenoma (GH4C1) pituicytes by inactivating the inhibitory GTP-binding protein Gi. AB - The phorbol ester 12-O-tetradecanoyl-phorbol 13-acetate (TPA) and thyroliberin exerted additive stimulatory effects on prolactin release and synthesis in rat adenoma GH4C1 pituicytes in culture. Both TPA and thyroliberin activated the adenylate cyclase in broken cell membranes. When combined, the secretagogues displayed additive effects. TPA did not alter the time course (time lag) of adenylate cyclase activation by hormones, guanosine 5'-[beta,gamma imino]triphosphate or forskolin, nor did it affect the enzyme's apparent affinity (basal, 7.2 mM; thyroliberin-enhanced, 2.2 mM) for free Mg2+. The TPA-mediated adenylate cyclase activation was entirely dependent on exogenously added guanosine triphosphate. ED50 (dose yielding half-maximal activation) was 60 microM. Access to free Ca2+ was necessary to express TPA activation of the enzyme, however, the presence of calmodulin was not mandatory. TPA-stimulated adenylate cyclase activity was abolished by the biologically inactive phorbol ester, 4 alpha-phorbol didecanoate, by the protein kinase C inhibitor polymyxin B and by pertussis toxin, while thyroliberin-sensitive adenylate cyclase remained unaffected. Experimental conditions known to translocate protein kinase C to the plasma membrane and without inducing adenylate cyclase desensitization, increased both basal and thyroliberin-stimulated enzyme activities, while absolute TPA enhanced adenylate cyclase was maintained. Association of extracted GTP-binding inhibitory protein, Gi, from S49 cyc- murine lymphoma cells with GH4C1 cell membranes yielded a reduction of basal and hormone-stimulated adenylate cyclase activities, while net inhibition of the cyclase of somatostatin was dramatically enhanced. However, TPA restored completely basal and hormone-elicited adenylate cyclase activities in the Gi-enriched membranes. Finally, TPA completely abolished the somatostatin-induced inhibition of adenylate cyclase in both hybrid and non-hybrid membranes. These data suggest that, in GH4C1 cells, protein kinase C stimulation by phorbol esters completely inactivates the n alpha i subunit of the inhibitory GTP-binding protein, leaving the n beta subunit functionally intact. It can also be inferred that thyroliberin conveys its main effect on the adenylate cyclase through activation of the stimulatory GTP-binding protein, Gs. PMID- 2569397 TI - Organelles involved in the intracellular transport of newly synthesized aminopeptidase N and their acidity. AB - The intracellular routes taken by aminopeptidase N, an apically expressed enzyme in the enterocyte, was followed in small intestinal cultures of pig using either immunoelectron microscopy (immunogold labeling) or continuous labeling with [35S]methionine. Aminopeptidase N was found in the microvillar membrane, the Golgi complex, apical small smooth vesicles, and various acidic lysosomal/endosomal-like organelles. By culturing mucosal explants in the presence of either cycloheximide or (3-(2,4-dinitroanilino)-3-amino-N methylpropylamine) (DAMP) it was demonstrated that the apical small smooth vesicles are exocytotic and that the low pH in the acid compartments is of no importance for intracellular transport and correct sorting of aminopeptidase N. Furthermore, our results show that the majority of the aminopeptidase N in the lysosomal/endosomal-like compartments is newly synthesized. PMID- 2569398 TI - Comparison of the effects of intravenous and oral betaxolol on antegrade and retrograde conduction in patients with atrioventricular nodal reentrant and atrioventricular reentrant tachycardia. AB - The electrophysiologic effects of intravenous (0.15 mg kg-1) and oral (20 mg day 1) betaxolol have been investigated in 11 patients with atrioventricular (A-V) nodal reentrant tachycardia and eight patients with orthodromic A-V reentrant tachycardia. Betaxolol significantly (P greater than 0.01) prolonged sinus cycle length, sinus node recovery time, intranodal conduction time, and the antegrade functional refractory period of the A-V node. When the effective refractory period of the A-V node could be determined it was increased by betaxolol, whereas no significant electrophysiologic effects were observed in the atrium, the ventricle or the accessory pathway. Intravenous betaxolol prevented tachycardia in 8 out of 11 patients with A-V nodal reentrant tachycardia, whereas oral betaxolol was effective in 10 patients, primarily by acting on the antegrade limb in two patients and on the retrograde limb in eight patients. In those with A-V reentrant tachycardia, intravenous betaxolol did not prevent tachycardia in any patient, while it was effective after oral treatment in two patients. When the tachycardia remained inducible, cycle length of the tachycardia increased in all patients, due to prolongation of the antegrade and retrograde conduction time in patients with A-V nodal reentrant tachycardia, and due to an increase in the antegrade conduction time, i.e. the A-V node, in the patients with A-V reentrant tachycardia. In conclusion, betaxolol proved to be effective in the treatment of supraventricular tachycardia; for chronic treatment, a single oral dose (20 mg) seems to suffice. PMID- 2569399 TI - Placental transfer and distribution of pinazepam and its metabolite N desmethyldiazepam in the maternal and fetal rabbit: effect of the stage of gestation. AB - The distribution of pinazepam and its metabolite N-desmethyldiazepam was studied in fetuses of New Zealand rabbits on the 20th and 27th day of pregnancy. The concentrations of both compounds were also measured in the maternal brain, liver and uterus. Pregnant rabbits were sacrificed at 0.5, 2, 4 and 12 h after intravenous administration of pinazepam (5 mg/kg). The concentrations of pinazepam and N-desmethyldiazepam in various biological specimens were measured by a specific gas-chromatographic procedure. Pinazepam and N-desmethyldiazepam rapidly crossed the placenta. In 20 day old fetuses, comparable concentrations of pinazepam were found in the liver, brain, heart, lungs and kidneys. In contrast, the liver of 27 day old fetuses accumulated pinazepam at concentrations higher than the other tissues. The hepatic extraction of pinazepam, already described in adult rabbits (1), develops prenatally. A preferential accumulation of pinazepam rather than N-desmethyldiazepam was also observed in the maternal uterus. In this tissue the concentrations of pinazepam were 5-10 times higher on the 27th rather than the 20th day of pregnancy. The stage of pregnancy influences the distribution pattern of pinazepam in rabbit fetuses and their mothers. PMID- 2569400 TI - Acute rejection of allogeneic hemopoietic progenitors by genetically resistant mice. AB - Natural resistance to hemopoietic allograft results in functional elimination of the graft from the host within 24 to 48 h. The resistance is specific and is directed to the cell surface target structures of an unidentified nature. These determinants are controlled by the major histocompatibility complex-linked hemopoietic histocompatibility (Hh) loci and expressed by cells of the hemopoietic system. In this study, the susceptibility of various hemopoietic progenitors, as well as the nature and the kinetics of the early rejection process, were examined by directly following the functional survival of the grafted progenitors by periodic sampling. In both F1 hybrid and inbred allogeneic hosts, multipotential progenitors for granulocyte-erythrocyte-monocyte megakaryocyte lineages, bipotential progenitors for granulocyte-monocyte lineages and unipotential progenitors of erythrocyte, granulocyte and monocyte were susceptible to elimination. Therefore, within the limited range of comparisons, neither the lineage, nor the degree of commitment or differentiation determines the susceptibility of hemopoietic progenitors to resistance. Functional elimination of the grafted progenitors from the recipient spleen was irreversible upon cultivation in semisolid medium. The frequency of seeding in the spleen at 3 h was comparable in resistant and susceptible hosts; elimination commenced shortly afterwards and by 24 h after transplantation less than 10% of clonogenic progenitors remained functional. Activation of natural killer cells strengthened pre-existing resistance, but did not convert genetically susceptible mice to resistant. Therefore, the resistance is dependent on an effector mechanism with predetermined specificity. PMID- 2569401 TI - A novel CD3-J11d+ subset of CD4+CD8- cells repopulating thymus in radiation bone marrow chimeras. AB - Sequential appearance of T cell subpopulations occurs in the thymus of irradiated AKR (H-2k, Thy-1.1) mice at an early stage after transplantation with bone marrow cells of C3H/HeN (H-2k, Thy-1.2) mice. The donor-derived thymocytes were first detected on day 8 after bone marrow reconstitution. Although most of the thymocytes were CD4-CD8- cells, an appreciable level of CD4+CD8- cells was detected in the thymus at this stage. The early appearing CD4+CD8- cells were a novel subset of thymocytes that were J11d+CD3-. From day 10 to day 21 the proportion of CD4+CD8-CD3-J11d+ cells decreased while the proportion of CD4+CD8+ cells and CD4+CD8-CD3+J11d- cells increased. The CD4+CD8-CD3- cells seem to diversify to form CD4+CD8+ thymocytes after short-term culture in vitro. These results suggested the existence of a differential pathway from CD4-CD8- cells to CD4+CD8+ cells via CD4+CD8- cells in thymus. PMID- 2569402 TI - Lymph node macrophage heterogeneity: the phenotypic and functional characterization of two distinct populations of macrophages from rat lymph node. AB - The isolation and characterization of lymph node macrophages (M phi) has shown a hitherto unknown heterogeneity. Two types of M phi were distinguished by morphology, monoclonal antibody staining and functional assays. The type I M phi failed to express surface Ia even when activated, a characteristic which has only previously been reported for splenic marginal zone M phi; despite studies suggesting an antigen presentation role for the M phi, the failure to express surface Ia would seem to eliminate an interaction with T helper cells for the type I M phi in the lymph node. In contrast, the type I M phi, other characteristics of clustering with activated B cells in vitro, the colocalization of the type I M phi and activated B cells in situ, the specific uptake of thymus independent type 2 antigens and the failure to undergo respiratory burst activity all suggest a M phi-B cell interaction, possibly of a trophic nature. The defective microbicidal activity of the type I M phi may have been compensated for by the type II M phi, which expresses both strong respiratory burst activity and surface Ia expression when freshly isolated. However, unlike the inflammatory M phi the activated phenotype of the type II M phi did not appear to be interferon gamma dependent because type II M phi could also be isolated from nude rat lymph nodes. PMID- 2569403 TI - Expression and ontogeny of murine CD2. AB - CD2 was first defined as the erythrocyte rosetting protein on the surface of human T cells. Recently, the rat and murine homologues have been identified by cDNA cloning. In this report we demonstrate that CD2 is expressed on the surface of most adult murine peripheral lymphocytes and thymocytes by indirect immunofluorescence using an anti-murine CD2 antiserum. The expression of CD2 on murine B cells was unexpected since in rat and human species it has been defined as a T cell-specific marker. Furthermore, CD2 appears very early on fetal thymocytes during development. The level of surface expression increases from day 13 of gestation to day 17, after which the surface density appears to reach a steady state. Thus, CD2 is expressed on day-13 thymocytes at the same stage that Thy-1, Pgp1 and the TcR gamma/delta/CD3 complex have been shown to be expressed. PMID- 2569404 TI - CD15 antibodies increase neutrophil adhesion to endothelium by an LFA-1-dependent mechanism. AB - Anti-neutrophil antibodies (CD15) bind to a simple sugar (lactose-N-fucopentaose III, LNF III) known to be present on the chains of the adhesion molecules on neutrophils. We have demonstrated that pre-incubation of neutrophils with a CD15 antibody increases neutrophil adherence to endothelium by a neutrophil-dependent mechanism. This augmented adhesion can be inhibited by antibodies directed against the alpha and beta chain of the leukocyte function-associated antigen 1 (LFA-1) molecule. There is also some increase in surface LFA-1 expression on neutrophils after CD15 incubation, suggesting that CD15 antibodies increase neutrophil adhesion by an LFA-1-dependent mechanism. The increase in LFA-1 expression after CD15 incubation occurs in the presence of a protein synthesis inhibitor. As LFA-1 is not stored intracellularly, the increased adherence of the neutrophils, and increased LFA-1 expression on their surface, suggests the possibility that the CD15 antibodies are binding to the LNF III antigen present on the alpha and beta chains of LFA-1, producing their effect by activating the molecule, perhaps by exposing new antigen sites by a stearic effect. PMID- 2569405 TI - Spinal beta-endorphin analgesia involves an interaction with local monoaminergic systems. AB - beta-Endorphin administered intrathecally (i.t.) in rats produced a dose dependent elevation in tail-flick latency. Naltrexone administered i.t. as a pretreatment reversed the spinal antinociceptive action of beta-endorphin, suggesting that the opioid interacts directly with spinal opiate receptors. Spinal administration of the alpha 1-adrenoceptor antagonist WB-4101 failed to alter the analgesic effects of the opioid, whereas the alpha 2-adrenoceptor antagonist yohimbine completely blocked beta-endorphin-induced elevations in tail flick latency. Thus, there is an apparent specificity for the alpha 2 adrenoceptor to mediate the spinal action of beta-endorphin. The 5-HT1 and 5-HT3 receptor antagonists (spiroxatrine and ICS 205-930, respectively) also reversed the analgesic effects of the opioid, while the 5-HT2 receptor antagonist ritanserin only partially blocked beta-endorphin-induced elevations in tail-flick latency. The present results suggest that beta-endorphin produces analgesia at the spinal level via an opiate receptor-mediated interaction with spinal monoaminergic nerve terminals. PMID- 2569406 TI - Amphetamine discrimination: effects of dopamine receptor agonists. AB - Dopamine (DA) D-1 and D-2 receptor agonists and antagonists were characterized in receptor binding and adenylate cyclase assays with respect to affinity, selectivity and efficacy. The ability of the ligands to interact with the discriminative stimulus effects of d-amphetamine (AMPH) was then assessed. The D 2 agonists, quinpirole, pergolide and CH 29-717, substituted completely for AMPH while neither partial (SKF 38393 and SKF 75670) nor full D-1 receptor agonists (SKF 89626 and SKF 81297) substituted. On the other hand, the selective D-1 and D 2 antagonists all blocked AMPH. The substitution for AMPH by pergolide was blocked by raclopride but not by SCH 23390, indicating D-2 mediation. In contrast, the motor effects of pergolide were blocked by both raclopride and SCH 23390, indicating mixed D-1/D-2 receptor involvement. These results suggest that D-1 and D-2 are equally involved in the expression of functional effects in the DAergic motor systems. Conversely, D-2 receptors may play a primary role in the DA systems involved in the AMPH cue; furthermore, the D-1 and D-2 receptors in the systems are relatively uncoupled. PMID- 2569407 TI - Differential effects of pimozide and SCH 23390 on acquisition of learning in mice. AB - Our main purpose was to clarify the differences between the effects of dopamine D 1 (SCH 23390) and D-2 (pimozide) antagonists on memory acquisition in a water finding and a one-trial passive avoidance task with ddY mice. In the water finding task, pimozide (0.1 and 0.2 mg/kg i.p.) enhanced the acquisition of latent learning of mice although it suppressed exploratory behavior. In contrast, SCH 23390 (0.1 mg/kg i.p.) attenuated the acquisition of latent learning and suppressed exploratory behavior. In the passive avoidance task, pimozide (0.1 and 0.2 mg/kg i.p.) enhanced the acquisition of the passive avoidance response of mice. SCH 23390 (0.05 and 0.1 mg/kg i.p.) failed to enhance the acquisition of the passive avoidance response. These results could suggest that a moderate block of D-2 receptors enhances memory acquisition but blocking D-1 receptors affects it in an opposite way. PMID- 2569408 TI - Effects of SK&F 85174, a DA-1/DA-2 receptor agonist, on pre- and postganglionic sympathetic neurotransmission to the heart. AB - We have performed experiments to determine the effects of SK&F 85174, a mixed DA 1/DA-2 receptor agonist, on the tachycardia elicited during pre- and postganglionic stellate stimulation in anesthetized dogs in order to identify a possible action of this compound on the stellate ganglia. SK&F 85174 produced hypotension and caused significant impairment of positive chronotropic responses elicited during pre- and postganglionic stellate stimulation. Pharmacological analysis of SK&F 85174-induced inhibition of cardiac sympathetic function with selective DA-1 and DA-2 receptor antagonists revealed that prior treatment with either S-sulpiride or domperidone (DA-2 receptor antagonists) significantly attenuated the inhibitory effects of SK&F 85174 on responses to pre- and postganglionic stellate stimulation. R-sulpiride (DA-1 receptor antagonist) failed to antagonize SK&F 85174-induced inhibition of tachycardia elicited during preganglionic stellate stimulation. Pretreatment with SCH 23390 (DA-1 receptor antagonist) did not modify the inhibitory effect of SK&F 85174 on responses to postganglionic nerve stimulation. However, SCH 23390 was most effective in antagonizing the hypotensive effect of SK&F 85174. These results show that SK&F 85174 inhibits sympathetic neurotransmission to the heart by activating presynaptic and possibly ganglionic DA-2 receptors, whereas the hypotension produced by SK&F 85174 results predominantly from the activation of the vascular DA-1 receptors. SK&F 85174 does not seem to exert any effect on the ganglionic DA receptors which are reported to be activated by the selective DA-1 receptor agonist, fenoldopam. PMID- 2569409 TI - Dopamine receptors in the stellate ganglion of the dog. AB - Effects of fenoldopam, a selective DA1 dopamine receptor agonist, and dipropyl dopamine and propylphenethyl dopamine, preferential DA2 dopamine receptor agonists, on ganglion transmission were studied in pentobarbital-anesthetized, open-chest dogs. Tachycardia induced by electrical stimulation (supramaximal voltage, 0.5 ms duration, 1-2 Hz) of the preganglionic cardio-accelerator nerves was monitored as a measure of ganglionic transmission. Drugs were injected into the costocervical artery (i.a.) close to the arterial supply of the ganglion. Doses required to produce 30-40% inhibition of ganglionic transmission by the i.a. route were 2-8 micrograms for dipropyl dopamine, 4-16 micrograms for propylphenethyl dopamine, and 100 micrograms for fenoldopam. At these doses none of the agonists affected tachycardia induced by electrical stimulation of the postganglionic nerve. Domperidone (5 micrograms/kg i.v.), a selective DA2 dopamine receptor antagonist, markedly antagonized the effects of dipropyl dopamine and propylphenethyl dopamine, but had only minor (and statistically insignificant) effects on the inhibitory effect of fenoldopam. SCH 23390 (5 micrograms/kg i.v.), a selective and potent DA1 antagonist, failed to modify the effects of any of the agonists. In a separate series, infusion of fenoldopam, 20 micrograms/kg per min i.v. for 5-7 min, facilitated postganglionic nerve stimulation and blocked the inhibitory effect of UK 14,304, an alpha 2 adrenoceptor agonist, on the postganglionic nerve. These results confirm and support the presence of DA2 but do not support the presence of the prototypal DA1 dopamine receptor in the mammalian ganglia. Furthermore, the alpha 2-adrenoceptor blocking property of fenoldopam points to the complication of using i.v. administration for studying its ganglionic actions while monitoring the target tissue effects in response to preganglionic nerve stimulation. PMID- 2569410 TI - The quinoxalinediones antagonise the visual firing of sustained retinal ganglion cells. AB - The non N-methyl-D-aspartate (NMDA) receptor antagonists dinitroquinoxaline-2,3 dione (DNQX) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), both inhibit the visually driven response of sustained ganglion cells in the cat retina in vivo. In contrast to these findings, the potent NMDA receptor antagonist 3-[+/- )-2 carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP) has no effect. Thus, the endogenous excitatory amino acid released onto these cells on visual stimulation acts at non-NMDA receptors. PMID- 2569411 TI - Intermittent neuroleptic treatment induces long-lasting abnormal mouthing in the rat. AB - Rats were treated either discontinuously or continuously with haloperidol. Only discontinuous treatment caused a long-lasting abnormal rise in the number of mouth movements. After termination of medication the animals were kindled electrically in the nucleus amygdala. The results suggest cross-sensitivity between discontinuous neuroleptic treatment and amygdala kindling. Our results are consistent with the hypothesis of a kindling-like sensitization to the dyskinetic-inducing side-effects of neuroleptic drugs following intermittent antidopaminergic treatment. PMID- 2569412 TI - Muscarinic agents modify kinetics properties of membrane-bound guanylyl cyclase activity. AB - Plasma membranes from bovine tracheal smooth muscle show guanylyl cyclase activity, which can be stimulated by muscarinic agonists such carbamylcholine and oxotremorine and blocked by atropine. This stimulation was observed in the presence of 150 mM NaCl. In the absence of this salt, guanylyl cyclase activity was considerably higher but was not affected by muscarinic agonists. Carbamylcholine decreased the apparent Km but did not change the Vmax of this enzyme. When plasma membrane fractions were extracted with 1% octylglucoside, guanylyl cyclase activity was preserved, however the muscarinic activation was abolished, despite a muscarinic receptor capable of [3H]quinuclidinylbenzilate binding being present in the extract. The detergent extraction changed the affinity of guanylyl cyclase for GTP but the Mn2+ kinetics was unaltered. Based on these findings and on current information in the literature, we propose that another component is required to restore the link between the muscarinic receptor and guanylyl cyclase, however the nature of this component remains to be established. PMID- 2569413 TI - [Comparative sensitivity of the sympathetic and parasympathetic ganglia to ganglionic blockaders]. AB - Pharmacological tests in vitro on the cat isolated superior cervical and ciliary ganglia identified the IEM-1194 as a selective parasympathetic ganglion-blocking agent. The ganglion-blocking activity of this drug was three times as high in parasympathetic ganglion (EC50 = 1.15 +/- 0.34 M/1 x 10(-5)) as in sympathetic one (EC50 = 3.2 +/- 0.2 M/1 x 10(-5)). I. v. administration of the IEM-1194 in a dose effectively (50%) blocking responses of the heart (bradycardia) and duodenum (peristaltics) to vagal stimulation, did not lower arterial blood pressure. PMID- 2569414 TI - Metformin potentiates B-cell response to high glucose: an in vitro study on isolated perfused pancreas from normal rats. AB - This study investigated the effects of metformin on pancreatic A-B- and D-cell functions using the isolated perfused rat pancreas model. The lactate output rate following metformin infusion was also monitored. Metformin was infused at the low "therapeutic" concentration of 1.5 micrograms/ml and its effects were evaluated in three different glycaemic conditions: during a basal infusion of 4.44 mM glucose, during a moderate increase to 8.88 mM of glucose concentration, and finally during a higher 16.66 mM glycaemic stimulus. Basal insulin secretion and B-cell release during the lower hyperglycaemic stimulus were unaffected by metformin infusion. On the contrary, the drug significantly enhanced insulin response to 16.66 mM glucose, particularly by increasing the second phase of hormone release. Glucagon and somatostatin releases during metformin infusion were similar to the secretory pattern observed in the control experiments both in the basal condition and in the presence of the two different hyperglycaemic stimuli. Finally metformin did not modify the lactate output rate from perfused pancreas, irrespective of the different glycaemic conditions employed. Therefore our data suggest--at least in rats, in in vitro experiments but above all in the presence of markedly elevated hyperglycaemic conditions--that metformin may influence the glucose stimulatory effect on B-cell activity. PMID- 2569416 TI - Study of serum copper and zinc in cases of hyperemesis gravidarum. AB - The dehydration, electrolyte and metabolic changes which occur in hyperemesis gravidarum are well recognized. The aim of this paper was to study the changes in serum copper and zinc and their correlation with the changes in serum electrolytes that occur in patients with hyperemesis gravidarum. Serum copper, zinc, sodium and potassium and urinary chloride were measured in 30 patients suffering from hyperemesis gravidarum and compared to the levels in 10 normal pregnant women in their first trimester. There was no significant change in serum copper or zinc in patients with hyperemesis compared to normal pregnant women. Also, there was no significant correlation between the changes in the level of these trace elements and the decrease which occurred in serum sodium and potassium and urinary chloride. PMID- 2569417 TI - Electronic fetal heart rate monitoring during cesarean section in cases of fetal distress. AB - Twelve fetuses in whom the diagnosis of fetal distress was made during first stage of labor were included in the study. All 12 patients underwent urgent cesarean section during which fetal heart rate (FHR) was recorded continuously. In 50% of the cases, marked improvement of FHR was registered early in the course of the operation. Improvement in fetal status is attributed to adequate oxygenation and cessation of uterine activity. PMID- 2569415 TI - Temporal changes in plasma and liver lipids and in the hepatic activities of acetyl-coenzyme a carboxylase and fatty acid synthetase after oestrogen treatment of the male chicken (Gallus domesticus). AB - 1. Male chickens (Gallus domesticus) were treated with a single intramuscular injection of oestradiol-17 beta, then changes in the liver and plasma levels of triacylglycerol, phospholipid, nonesterified fatty acids and in the hepatic activities of acetyl-CoA carboxylase and fatty acid synthetase were measured at various times after injection. 2. The results suggest that the initial phase (less than 20 hr) of oestrogen-induced hyperlipidaemia occurs in the absence of changes in the hepatic activities of the major enzymes of fatty acid biosynthesis, but a subsequent increase in these enzyme activities may contribute to the later phase (greater than 20 hr) of oestrogen-induced lipogenesis in avian liver. PMID- 2569418 TI - Observations on molar pregnancy in Enugu, Nigeria. AB - Forty-one molar pregnancies seen at the University Teaching Hospital Enugu, Nigeria during the 10-year period 1976-1985 are analyzed. The incidence of hydatidiform mole was 0.82 per 1000 pregnancies. The incidence of molar pregnancy was lowest among teenage women and increased markedly with advancing age over 35 years. Hyperemesis gravidarum, a relatively uncommon finding in Nigerian women during normal pregnancy was a prominent feature of molar pregnancy in the women. The incidence of postmolar gestational trophoblastic disease was 17%; choriocarcinoma occurred in 7% of the subjects. Molar pregnancy and postmolar malignant trophoblastic disease occurred much less frequently among the Igbo women of Enugu than the Yorubas of western Nigeria. Earlier and better recognition and treatment of hydatidiform mole will probably result in a decrease in the high rate of complications found during this study. PMID- 2569420 TI - Hydatidiform mole in the elderly: hysterectomy or evacuation? AB - Ten patients with hydatidiform mole treated by primary hysterectomy were retrospectively compared with 24 patients whose moles were evacuated by suction curettage. All patients were 35 years old or more and were followed up for 1 year following the evacuation. During follow-up, 10% of the hysterectomized patients needed chemotherapy in comparison with 33.4% of the non-hysterectomized patients. This difference was not statistically significant. There was also no statistically significant difference when the serial serum beta-subunit of human chorionic gonadotrophin (beta-hCG) regression rates of the two groups were compared. The study shows that primary hysterectomy does not worsen the prognosis of gestational trophoblastic disease, however, it does not negate the need for careful follow-up. PMID- 2569419 TI - Bari district birthweight study: comparison with the Bristol perinatal growth chart. AB - We studied the neonatal weight distribution of 15,298 newborns from the Bari District at various gestational ages by the 10th, 50th, and 90th percentiles. After analysis, we constructed a growth chart following the method of Dunn. By comparing our results with those of Dunn's Bristol Perinatal Growth Chart, we found a weight difference of +4.5% in our population. This difference may be related to ethnic as well as socioeconomic factors. PMID- 2569421 TI - Enhanced pre-ovulatory progesterone levels in fertile cycles during clomiphene citrate treatment. AB - We compared ovulatory changes in fertile and preceding infertile cycles in 21 patients with unexplained infertility conceiving after clomiphene citrate treatment. No significant differences were observed in follicular growth, cervical score and follicle stimulating hormone (FSH) levels. Progesterone was higher (P less than 0.05) in the 2 days preceding ovulation in fertile cycles, luteinizing hormone (LH) higher (P less than 0.05) the day before, and 17-beta estradiol lower (P less than 0.05) 4 days before. Stimulating progesterone secretion by systematic LH administration before ovulation could improve secretory endometrial transformation and thus reproductive prognosis. PMID- 2569422 TI - Interval insertion of an intrauterine contraceptive device following cesarean section. AB - Two previous studies of interval insertion of intrauterine contraceptive devices (IUCD) in women with previous cesarean delivery produced conflicting results. We studied the first and subsequent segment IUCD performance in 215 women with one or more cesarean deliveries. All the insertions were performed by doctors in the outpatient department of the hospital. The follow-up rate at 6 months was over 80%. There was a higher than expected expulsion rate of 19 per 100 insertions in the first 12 months, in both the first and subsequent segments. Other IUCD related problems were in keeping with expectations. The IUCD nonetheless remains a useful contraceptive option for these women. PMID- 2569424 TI - Primary hyperparathyroidism and pregnancy. PMID- 2569423 TI - Transient blindness in pregnancy induced hypertension. AB - Transient blindness associated with pregnancy induced hypertension without neurological symptoms is a rare phenomenon. The blindness in these cases is postulated to be of "cortical blindness". Two such cases occurring immediately after childbirth are presented. The ophthalmic manifestations of this complication of pregnancy induced hypertension and the importance of an urgent evaluation are discussed. PMID- 2569425 TI - Pneumoperitoneum due to peritoneovaginal fistula following hysterectomy. AB - A patient with pneumoperitoneum that developed due to peritoneovaginal fistula following hysterectomy is presented. In addition, we have reviewed the literature and formulated guidelines for the management of this problem. Most patients present following sexual intercourse and in many cases the fistula can be diagnosed radiographically. Laparotomy may be avoided if the cause of the fistula is suspected. Surgical closure of the fistula may be unnecessary since spontaneous closure occurs frequently. PMID- 2569426 TI - Carcinoma of the uterine cervix and schistosomiasis. AB - The relation between cervical cancer and schistosomal infection is controversial. A case of well differentiated adenocarcinoma associated with schistosomiasis of the cervix is presented. A 45-year-old female complaining of intermenstrual bleeding had a polypoid ulcerated cervical mass. Biopsy revealed well differentiated adenocarcinoma and bilharzial ova with terminal spine embedded in the tumor tissue. The relation between cancer cervix and schistosomiasis is discussed. PMID- 2569427 TI - Use of prostaglandin E2 vaginal suppositories in third-trimester fetal demise. AB - A retrospective analysis of eight cases of third-trimester fetal demise managed with prostaglandin E2 vaginal suppositories (PGE2) is presented. Management included laminaria insertion prior to induction and an initial lower dosage of PGE2. No cases of uterine rupture or cervicovaginal lacerations were encountered. A summary of the literature as it relates to PGE2 vaginal suppository use in third-trimester fetal demise is included. PMID- 2569428 TI - Ionophore bromo-A23187 reveals cellular calcium stores in single pituitary somatotropes. AB - Free cytosolic calcium concentration, [Ca2+]i, in single rat pituitary cells can be measured with the fluorescent, calcium-sensitive probe fura-2 and digital image analysis. A reverse hemolytic plaque assay (RHPA) identifies somatotropes in the mixed population of pituitary cells. Previous studies showed that growth hormone releasing factor (GRF) stimulates growth hormone (GH) release from pituitary somatotropes by increasing the influx of calcium into the cell. Somatostatin reduced [Ca2+]i and inhibits hormone release presumably by closing calcium channels in the membrane. The calcium-ionophore bromo-A23187 rapidly increased [Ca2+]i from a baseline of 226 +/- 38 nM to a peak of 842 +/- 169 nM (mean +/- SEM) which was reached 30 s after exposure to the drug. This spike was followed by a sustained phase of elevated [Ca2+]i approximately 370 nM. When somatostatin (SRIF) (10 nM) was combined with ionophore treatment, the initial rise was preserved. However, the second phase was abolished and SRIF lowered [Ca2+]i to 57 +/- 7 nM. Depolarizing the cellular membrane with high extracellular potassium (60 mM) increased cytosolic calcium as well (797 +/- 178 nM); however, this was not affected by the addition of SRIF (988 +/- 71 nM). KCl depolarization in calcium-free medium (+1.5 mM EGTA) provoked no rise in cytosolic calcium. In contrast, after ionophore, the initial spike was preserved while the sustained phase of elevated [Ca2+]i was abolished. We conclude from these data that (1) membrane depolarization and ionophore treatment lead to an influx of calcium into the cytosol of normal pituitary somatotropes. (2) SRIF inhibits calcium influx induced by ionophore but not influx after depolarization with high potassium concentrations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569429 TI - DNA transfection into mammalian cells, selective markers and gene cointegration. AB - The dominant selective markers that we and others have constructed for the selection of cells which stably acquired a foreign DNA, are briefly described. The possibility of obtaining the cointegration of several genes, and of inducing their amplification in the recipient cell, is discussed. PMID- 2569431 TI - Characterization of a pilus produced by Aeromonas hydrophila. AB - A pilus produced by Aeromonas hydrophila was purified and partially characterized. The pilin monomers had an apparent molecular weight of 17,000. Agglutination studies indicated serological cross-reactivity in the pili of A. hydrophila strains. Presence of pili did not correlate with hydrophobicity or haemagglutinating ability of the bacteria. PMID- 2569430 TI - Analysis of the HepG2/erythrocyte glucose transporter locus in a family with type 2 (non-insulin-dependent) diabetes and obesity. AB - A recent report has shown an association between a specific Xba1 restriction fragment of the human HepG2-Erythrocyte glucose transporter gene and Type 2 (non insulin dependent) diabetes. To further examine the significance of this finding we have studied Type 2 diabetic pedigrees for linkage between the Xba1 alleles of this glucose transporter gene and diabetes. One large pedigree, in which the diabetic phenotype was associated with obesity and insulin resistance, was informative. In this family the disease did not co-segregate with the glucose transporter locus. Formal linkage analysis was performed assuming autosomal dominant inheritance with age-dependent penetrance. At putative gene frequencies of 0.01 and 0.001 the logarithm of the odds for linkage versus non-linkage at a recombination fraction of 0.001 was -1.84 and -3.32 respectively (a value of less than -2 indicates definite non-linkage). Genetic variations in the HepG2 Erythrocyte glucose transporter gene are unlikely to be responsible for the development of diabetes in this pedigree. PMID- 2569432 TI - Extragenic suppression and synthetic lethality among Chlamydomonas reinhardtii mutants resistant to anti-microtubule drugs. AB - The antimicrotubule agents oryzalin (ORY), colchicine (COL) and taxol (TAX) were used to select three recessive, conditional lethal (ts-) mutants which defined two new essential loci, ory1 and cor1. The two ory1 mutants conferred resistance to ORY, TAX, and COL; the cor1 mutant conferred resistance only to COL. Each of the mutants displayed wild-type sensitivity to a number of unrelated inhibitors. Assembly and disassembly of flagellar microtubules in the ory1 mutants displayed wild-type sensitivity to ORY and COL, suggesting that the ory1 gene product either does not participate in these processes or the ory1 gene product alone is not sufficient to confer resistance. The ory1 locus mapped to linkage group X; cor1 was mapped to the left arm of linkage group XII. A synthetic lethal interaction was observed between ory1 and cor1 mutations, i.e., inferred ory1 cor1 double mutants were inviable at the permissive temperature. The conditional lethal phenotype of ory1-1 was used to select many spontaneous TS+ revertants, which arose at high frequencies. Genetic and phenotypic characterization of the revertants demonstrated that (1) the revertants fell into four phenotypic classes, including some which conferred supersensitivity to ORY and others which conferred cold-sensitive lethality, (2) reversion was caused in most or all cases by extragenic suppressors, (3) suppressor mutations displayed complex behavior in heterozygous (sup/+) diploids, (4) many different loci may be capable of suppressing ory1 mutants, and (5) suppressors of ory1-1 efficiently suppressed an independently isolated allele, ory1-2. Taken together the ory1, cor1, and suppressor mutations identify a number of interacting loci involved in essential cellular processes which are specifically susceptible to antimicrotubule agents. PMID- 2569433 TI - Reciprocal recombination and the evolution of the ribosomal gene family of Drosophila melanogaster. AB - The role of reciprocal recombination in the coevolution of the ribosomal RNA gene family on the X and Y chromosomes of Drosophila melanogaster was assessed by determining the frequency and nature of such exchange. In order to detect exchange events within the ribosomal RNA gene family, both flanking markers and restriction fragment length polymorphisms within the tandemly repeated gene family were used. The vast majority of crossovers between flanking markers were within the ribosomal RNA gene region, indicating that this region is a hotspot for heterochromatic recombination. The frequency of crossovers within the ribosomal RNA gene region was approximately 10(-4) in both X/X and X/Y individuals. In conjunction with published X chromosome-specific and Y chromosome specific sequences and restriction patterns, the data indicate that reciprocal recombination alone cannot be responsible for the observed variation in natural populations. PMID- 2569434 TI - Human serum transglutaminase and coeliac disease: correlation between serum and mucosal activity in an experimental model of rat small bowel enteropathy. AB - Transglutaminase (TG) activity is increased in the mucosa of patients with coeliac disease. Among 18 patients with untreated coeliac disease we have found a significant decrease (p less than 0.001) in serum levels of TG activity (0.72 (0.23) mU/ml). There was no significant differences between 16 treated coeliacs (1.24 (0.28) mU/ml) and 30 normal controls (1.63 (0.42) mU/ml). To evaluate the connection between serum and mucosal TG activity we used the experimental model of methotrexate induced acute hypoplastic enteropathy in the rat. Transglutaminase activity was unchanged in serum and mucosa 24 and 48 hours after MTX administration, but increased in mucosa (2.606 (0.95) v basal 0.207 (0.026) mU/mg protein, p less than 0.001) and significantly decreased in serum at 72 hours (2.08 (0.38) v basal 5.56 (1.50) mU/ml, p less than 0.001) during intestinal cell proliferation. Activity of the enzyme in the mucosa and serum returned to baseline levels within 120 hours. This experimental animal model helps to explain the data of TG activity in human intestinal mucosa and serum reported in this study. Results are mean (SD). PMID- 2569435 TI - Comparison of orocaecal transit times assessed by the lactulose/breath hydrogen and the sulphasalazine/sulphapyridine methods. AB - The lactulose/breath hydrogen and the sulphasalazine/sulphapyridine methods of assessing orocaecal transit time have been compared. In a two part crossover study in healthy normal subjects the median orocaecal transit time by the SLZ/SP method was 4.84 hours but only 2.92 hours by the lactulose/breath hydrogen method. Coadministration of lactulose and sulphazalazine to nine subjects with assessment of orocaecal transit time by hydrogen breath determination and plasma sulphapyridine assay gave orocaecal transit times of 2.33 and 2.25 hours respectively suggesting that the lactulose reduces transit time and that the lactulose/breath hydrogen method, which is so convenient to use, gives artificially low transit times. A third experiment was undertaken to compare the orocaecal transit times after 1.5 and 3.0 g sulphazalazine. The orocaecal transit times after the two doses were not statistically different. PMID- 2569436 TI - Sudden death from asthma. AB - Deaths from asthma investigated by the Department of Forensic Medicine, The Gade Institute, University of Bergen during the period 1977-1986 were recorded. There were 11 cases with information in the police report of the use of pressurised bronchodilating aerosols prior to death. Seven of the victims were found with the aerosol in the hand or close by the body. Overdosage due to excessive inhalation were suspected in several of the victims. The possibility of side effects due to overuse is discussed. Other mechanisms of death are also discussed. PMID- 2569437 TI - Production of discrete high specific activity DNA probes using the polymerase chain reaction. AB - Conditions are described for the synthesis of discrete DNA probes with high specific activity using the polymerase chain reaction. This method enables the production of DNA probes between any two oligonucleotide sequences from cloned or uncloned templates. These probes are uniform in length and their specific activity (1 x 10(9) cpm/microgram) is comparable to probes produced by other methods. PMID- 2569438 TI - Understanding depression in schizophrenia. PMID- 2569439 TI - Characterisation of Escherichia coli adhesins in patients with symptomatic urinary tract infections. AB - The ability of bacteria to adhere to the epithelial cells of hosts has been shown to be mediated by adhesins. Many of these show readily demonstrable haemagglutinating activity. Of 109 Escherichia coli strains isolated from patients with symptomatic urinary tract infection, 11 (10.1%) were identified by their haemagglutinating properties as being P fimbriated, which was confirmed by the latex bead test. Other classes of adhesins, termed X and "other", were found in mannose resistant haemagglutinating E coli strains, which represented 4.6% (5) and 0.9% (1), respectively, of all the strains. Type 1 fimbriae were found in 40.4% (44/109) of E coli strains grown on colonising factor agar (CFA) medium. This incidence was 12.8% higher (53.2%, 58/109) when the strains were grown on CFA supplemented with urea, which suggested that urea may modulate the expression of type 1 fimbriae. Conversely, this phenomenon was not seen in P fimbriated E coli. Assays using trypsinised and non-trypsinised human erythrocytes showed no difference in the percentage of strains that haemagglutinated. Regarding the clinical correlation of fimbriated E coli strains, the X mannose resistant haemagglutinating adhesins were also found to be of clinical relevance. P fimbriated E coli strains were isolated from five out of the eight patients with pyelonephritis. PMID- 2569440 TI - Correlation between RsaI restriction fragment length polymorphism and electrophoretic types of human placental alkaline phosphatase. AB - Restriction fragment length polymorphism (RFLP) of human alkaline phosphatases was studied in a population sample from northern Sweden using a placental alkaline phosphatase (PLAP) cDNA probe. After digestion of human genomic DNA with RsaI the Southern blots showed DNA fragments most probably derived from three genes: PLAP, germ cell alkaline phosphatase (PLAP-like) and intestinal alkaline phosphatase. In agreement with a previous study, a two-allele polymorphism was found in PLAP with bands at 1.6 kilobases (A1) and 1.8 kilobases (A2). The gene frequencies of A1 and A2 were 0.46 and 0.54, respectively. There was a significant correlation between the RsaI RFLPs and electrophoretic types of PLAP; RSAI A2 showed an association with the ALP2p allele of PLAP. PMID- 2569441 TI - Role of cloned virulence factors (mannose-resistant haemagglutination, mannose resistant adhesions) from uropathogenic Escherichia coli strains in the release of inflammatory mediators from neutrophils and mast cells. AB - Genetically cloned E. coli strains expressing cloned virulence factors were studied with regard to their capability to induce inflammatory mediator release from various target cells. Among the strains were E. coli strains with mannose resistant haemagglutination (MRH+) and mannose-resistant adhesins, e.g. E. coli 536/21 pANN 801/4, E. coli 536/21 pANN 921 and E. coli 536/21 pANN 801-1. In comparison, E. coli 536/21, E. coli 536/21 pGB 30 int and E. coli K12, without and with mannose-sensitive haemagglutination (MSH +/-), and adhesins were studied. The properties of the various strains for human PMN with regard to adherence and phagocytosis, chemiluminescence, 5-lipoxygenase activation of arachidonic acid, leukotriene formation, granular enzyme release and release of histamine from rat mast cells were analysed. It is evident that the various biochemical processes of cell activation are dissociated events. The highest chemiluminescence response is obtained with strains expressing MSH+, P-MRH+ or S MRH+; the presence of S-adhesins suppressed the response. Highest leukotriene formation is obtained with E. coli 536/21 pANN 801-4, while E. coli with MSH was inactive. The concomitant presence of haemolysin secretion enhanced mediator release significantly. Our data suggest a potent role for mannose-resistant haemagglutination (MRH), adhesins and haemolysin as virulence factors in inducing the release of inflammatory mediators. PMID- 2569442 TI - The D17Tu5 locus in the t complex: implications for the origin of t haplotypes and inbred strains. AB - The mouse x Chinese hamster cell line R4 4-1 contains only one mouse chromosome, the bulk of which corresponds to Mus musculus chromosomes 17 and 18 (MMU17 and MMU18, respectively). A genomic library was prepared from the R4 4-1 DNA, and a mouse clone was isolated from the library, which-with the help of somatic cell hybrids-could be mapped to the MMU17. A locus defined by a 2.7-kb long Bam HI probe from this clone was designated D17Tu5 (Tu for Tubingen). The locus proved to be polymorphic among inbred strains and wild mice. By testing of recombinant inbred strains and partial t haplotypes, the D17Tu5 locus could be mapped to a position between the D17Leh66E and D17Rp17 loci within the t complex. Two alleles were found at this locus, D17Tu5a and D17Tu5b, defined by Taq I restriction fragment length polymorphism. Both alleles are present among inbred strains and wild mice of the species M. domesticus. All complete t haplotypes tested carry the D17Tu5a allele and all tested wild mice of the species M. musculus, with the exception of those bearing t haplotypes, carry the D17Tu5b allele. Additional alleles are found in some populations of wild mice and in other species of the genus Mus. The distribution of the two alleles among the inbred strains correlates well with their known or postulated genealogy. Their distribution between the two species of Mus and among the mice with T haplotypes suggests a relatively recent origin of the t haplotypes. PMID- 2569443 TI - HLA-DQw7 is a disease severity marker in patients with rheumatoid arthritis. PMID- 2569444 TI - Unusual HLA-DR/DQ haplotypes: two different breakpoints in two different DR2-DQw3 haplotypes. PMID- 2569445 TI - New evidence for trans-species evolution of the H-2 class I polymorphism. AB - A serological survey using alloantisera specific for the H-2 class I antigens in Japanese wild mice. Mus musculus molossinus, revealed a high frequency of the H 2Kf antigen. This antigen has also been found in European wild mice, M. m. domesticus and M. m. musculus. In this survey, the H-2Kf antigen was characterized through the use of ten newly isolated monoclonal antibodies raised against cells of a Japanese wild mouse, and by Southern blot analysis using an H 2K locus-specific probe which hybridizes with the 3' end of the gene. The serologically identified H-2Kf antigens revealed several minor variations in reactivities to the monoclonal antibodies. However, all the antigens examined could be clearly separated into two types with respect to the restriction fragment length polymorphism (RFLP) pattern. The first type, found together with a single, characteristic RFLP pattern, was always associated with the presence of reactivity to one particular monoclonal antibody MS54. The second type, found to represent different RFLP patterns, is associated with the absence of reactivity to MS54. This concordance between the presence of an antigenic determinant and a particular RFLP was observed not only within Mus musculus subspecies but also in a different species: M. spretus, carrying the same antigenic determinant, gave an identical RFLP to that of the other MS54-positive Mus musculus subspecies. The data suggest that the antigenic determinant specific for MS54 is an ancient polymorphic structure which has survived the long period of diversification of Mus species (approximately 2-3 million years) without alteration, and is associated with a stable DNA structure at the 3' end of the H-2K gene. PMID- 2569446 TI - Somatostatin inhibition of fructose-induced hypertension. AB - The role of insulin resistance and hyperinsulinemia in the etiology of fructose induced hypertension was studied in male Sprague-Dawley rats. Rats consumed a fructose-enriched diet (containing 66% of total calories as fructose) for 11 days and were infused continuously during the last 7 days with either a somatostatin analogue or vehicle. At the end of this period, rats receiving the somatostatin analogue had a lower plasma insulin concentration (52 +/- 4 vs. 70 +/- 6 microunits/ml, p less than 0.01) and a lower blood pressure (133 +/- 2 vs. 150 +/ 2 mm Hg) than did the rats infused with the control solution. In addition, the increase in plasma triglyceride concentration in response to the fructose enriched diet was significantly attenuated (p less than 0.001) in the rats infused with somatostatin. These data provide further support that the increase in blood pressure that occurs when normal rats are fed a high fructose diet is dependent on the ability of this intervention to cause insulin resistance and hyperinsulinemia. PMID- 2569447 TI - Use of the promoter fusion transposon Tn5 lac to identify mutations in Bordetella pertussis vir-regulated genes. AB - Mutants of Bordetella pertussis deficient in virulence-associated factors were identified by using the transposon Tn5 lac. Tn5 lac is a derivative of Tn5 which generates promoter fusions for beta-galactosidase. Tn5 lac insertions in the vir regulated genes of B. pertussis were identified by selecting for kanamycin resistant mutants that expressed beta-galactosidase when the vir-regulated genes were expressed but not when the vir-regulated genes were turned off. Fourteen different mutations in vir-regulated genes were identified. Two mutants were deficient in the production of the filamentous hemagglutinin, two mutants were deficient in the production of adenylate cyclase toxin and hemolysin, and one mutant was deficient in the production of dermonecrotic toxin. One insertion mapped adjacent to the pertussis toxin gene, but the mutant produced pertussis toxin. The phenotypes of the remaining eight mutants were not determined, but the mutants did not appear to be deficient in the production of the 69,000-dalton outer membrane protein (agglutinogen 3) or the capsule. Screening for mutations in either of the fimbrial genes proved to be problematic since the parental strain was found to switch from a fimbriated to a nonfimbriated state at a high frequency, which was suggestive of the metastable expression of pili in other bacteria. We used Southern blot analysis with a 30-mer specific for the fimbrial sequences. No bands with the predicted increase in size due to the 12 kilobases from Tn5 lac were observed, which suggests that none of these genes were mutated. Southern blot analysis also revealed that seven of the eight unidentified mutations mapped to different restriction fragments, which suggests that they could be deficient in as many as seven different genes. PMID- 2569448 TI - Cross-reactivity of Pseudomonas aeruginosa antipilin monoclonal antibodies with heterogeneous strains of P. aeruginosa and Pseudomonas cepacia. AB - Much of the morbidity and mortality in patients with cystic fibrosis (CF) is secondary to pulmonary infections with Pseudomonas aeruginosa and, more recently, with Pseudomonas cepacia. Prevention of colonization and subsequent infection would be a useful therapeutic strategy. The pili (fimbriae) of P. aeruginosa are a potential vaccine antigen, as they have been implicated in binding to respiratory epithelium and appear to have limited antigenic diversity. Monoclonal antibodies (MAbs) raised to P. aeruginosa pilin demonstrated significant cross reactivity, as four of five P. aeruginosa strains with known pilin sequences and 10 of 15 P. aeruginosa clinical isolates hybridized by immunoblot with at least one of the three MAbs tested. The P. cepacia strains demonstrated minimal cross reactivity with these MAbs, as only 2 of 16 strains hybridized immunologically. The three MAbs decreased the adherence of 35S-labeled P. aeruginosa PA1244 to bovine tracheal cells by 56, 45, and 31%. One of these MAbs decreased the adherence of strains P. aeruginosa PAO1 and P. cepacia 249 to CF epithelial cells by 46 and 25%, respectively. While antibodies to Pseudomonas pili must be shown to be protective in patients with CF, these studies give support for a multivalent vaccine strategy using P. aeruginosa pilin as the immunogen. PMID- 2569449 TI - Fimbria-associated adhesin of Bacteroides loeschei that recognizes receptors on procaryotic and eucaryotic cells. AB - Inhibition studies with a set of adhesin-specific monoclonal antibodies and various sugars revealed that a fimbria-associated adhesin of Bacteroides loeschei recognizes receptors on both procaryotic and eucaryotic cells. These interactions permit this bacterium to attach to both types of cells, producing coaggregates in the presence of strains of Streptococcus sanguis and hemagglutination in the presence of neuraminidase-treated human erythrocytes. PMID- 2569450 TI - [Experiences with a new fat emulsion in surgical intensive care medicine]. AB - As far as energy availability is concerned, it is advantageous to give lipid emulsions to severely catabolic patients, in comparison to lipid-free TPN. It is important to administer essential fatty acids, especially linoleic and linolenic acid, which play a major part in synthesis of membrane phospholipids and prostaglandin metabolism. Lipid emulsions with linoleic acid content and high linoleic-linolenic ratio might be of great value in posttraumatic situations, where the need for linoleic acid might be increased up to 50 g/day. We have examined the safety of a new lipid emulsion with a linoleic-linolenic acid ratio of 16:1 in 14 severely catabolic surgical ICU patients. As a result, neither liver enzyme elevation nor allergic reactions occurred. The limited elevation of triglycerides under lipid infusion and the rapid fall of these values to normal levels account for the satisfactory utilization of the examined lipid emulsion in critically ill patients. PMID- 2569451 TI - [Studies of solutions with various branched-chain amino acid contents on protein metabolism in the postoperative period. 1. A 15N tracer kinetic animal model for evaluating the effect of parenteral amino acid administration on nitrogen metabolism]. AB - We were able to estimate nitrogen or protein metabolism on the basis of a 15N metabolic model that delivered a high amount of data during a parenteral nutrition period over 72 h. We describe the basic principles of the necessary N or 15N-analyses in blood, as well as in sera, in urine, in tissues of selected organs and in hepatocytes. This 15N-metabolic model allows the estimation of the utilization of amino acid infusion solutions. Because of the great similarity of parameters of the N or protein turnover between man and pig it is possible to provide relevant information for the clinical practice regarding amino acid infusion solutions. PMID- 2569452 TI - Neurochemicals and respiratory control during development. AB - During ontogeny, the central nervous system undergoes neuronal growth, regression, and remodeling. The development of neurotransmitter and modulator systems is a plastic process with individual temporal characteristics for each system. These characteristics include the synthesis, degradation, or uptake of neurochemicals and, largely independently, the appearance of their receptors. Message transmission during ontogeny is compounded by the variable development of these systems and by the coexistence and cofunction among these chemicals. Nine neurochemical systems are discussed: adenosine, gamma-aminobutyric acid, opioids, prostaglandins, serotonin, progesterone, substance P, thyrotropin-releasing hormone, and the catecholamines. The possible role of each of these in natural perinatal respiratory control is evaluated according to predetermined criteria. These include the presence of a substance system in respiratory-related regions, physiologically appropriate changes in its concentration in these regions, elicitation of respiratory effects by agonists and antagonists, and abolition with an antagonist of the effect of a substance during its presumed activation by a physiological process. It is suggested that excessive levels of suppressant neuromodulators or an imbalance among neurochemicals can partly explain the special features of respiratory control in the perinatal period. PMID- 2569453 TI - Dopamine and ventilatory effects of hypoxia and almitrine in chronically hypoxic rats. AB - We hypothesized that the temporary blunted ventilatory response to hypoxia seen in chronically hypoxic rats could be related to the increased amount of dopamine found in their carotid bodies. Rats, kept 2-3 wk in 10% O2, showed reduced nonisocapnic ventilatory responses to 21-12% inspiratory O2 fraction compared with control rats. Stimulus-response curves to almitrine, which simulates the action of hypoxia on the carotid body, were also depressed in chronically hypoxic rats. Responses to hypoxia and almitrine were significantly correlated in the two groups of rats. Dopamine depressed ventilation during normoxia, hypoxia, and almitrine stimulation in both groups, an action abolished by the dopamine-2 antagonist domperidone. Domperidone slightly increased responses to hypoxia and almitrine in control rats but had a greater enhancing effect in chronically hypoxic rats, such that there was no longer a difference between the responses of the two groups. PMID- 2569455 TI - Analysis of responses to sympathetic nerve stimulation in the feline pulmonary vascular bed. AB - The adrenergic receptor subtypes mediating the response to sympathetic nerve stimulation in the pulmonary vascular bed of the cat were investigated under conditions of controlled blood flow and constant left atrial pressure. The increase in lobar vascular resistance in response to sympathetic nerve stimulation was reduced by prazosin and to a lesser extent by yohimbine, the respective alpha 1- and alpha 2-adrenoceptor antagonists. Moreover, in animals pretreated with a beta-adrenoceptor antagonist to prevent an interaction between alpha- and beta 2-adrenoceptors, responses to nerve stimulation were reduced by prazosin, but yohimbine had no significant effect. On the other hand, in animals pretreated with a beta-adrenoceptor antagonist, yohimbine had an inhibitory effect on responses to tyramine and to norepinephrine. Propranolol had no significant effect on the response to nerve stimulation, whereas ICI 118551, a selective beta 2-adrenoceptor antagonist, enhanced responses to nerve stimulation and injected norepinephrine. The present data suggest that neuronally released norepinephrine increases pulmonary vascular resistance in the cat by acting mainly on alpha 1-adrenoceptors and to a lesser extent on postjunctional alpha 2 adrenoceptors but that this effect is counteracted by an action on presynaptic alpha 2-receptors. The present studies also suggest that neuronally released norepinephrine acts on beta 2-adrenoceptors and that the response to sympathetic nerve stimulation represents the net effect of the adrenergic transmitter on alpha 1-, alpha 2-, and beta 2-adrenoceptors in the pulmonary vascular bed. PMID- 2569454 TI - Stimulus interaction between CO2 and almitrine in the cat carotid chemoreceptors. AB - The hypothesis that augmentation of the carotid chemoreceptor response to hypoxia by almitrine is due in part to an increased response to CO2 was tested by using single or few fiber preparation of carotid body chemosensory fibers in 12 cats anesthetized with alpha-chloralose. To differentiate between the plausible mechanisms of effects, we also tested the responsiveness of the afferents to cyanide and nicotine before and after almitrine. After a saturation dose of almitrine (1 mg.kg-1 followed by 0.5 mg.kg-1.h-1) the chemosensory responses to CO2 strikingly increased even during hyperoxia: the afferents showing an increased transient peak activity at the onset of hypercapnia, an augmented steady-state response to CO2 stimulus, and a decreased arterial PCO2 stimulus threshold. Thus, the effect of almitrine on carotid chemoreceptor response to hypoxia could be explained, at least in part, by its multiplicative stimulus interaction with CO2. After almitrine, the chemoreceptor response to cyanide, which is dependent on arterial PO2, was not particularly augmented relative to those of nicotine. Accordingly, the O2-sensing mechanism does not appear to be the primary site of almitrine effect. The results also indicate that the site of CO2 chemoreception resides downstream from those of hypoxia. PMID- 2569456 TI - Alcohol withdrawal: a review of clinical management. AB - The authors review recent developments in the management of alcohol withdrawal, including its clinical components, available treatment strategies, and some recent neurochemical and endocrine research in the area. PMID- 2569457 TI - Twenty neuroleptic rechallenges after neuroleptic malignant syndrome in 15 patients. AB - Neuroleptic medication was reintroduced to 15 patients who had suffered an episode of neuroleptic malignant syndrome (NMS). Thirteen (87%) were eventually able to take neuroleptics again, indicating that they did not have an inherent intolerance. The authors analyzed the ability of patients to tolerate rechallenge according to the time of reintroduction after resolution of a previous episode and the relative neuroleptic potency and dosage compared with the initial episode. Successful rechallenge was highly significantly related to the time that had elapsed following resolution of the previous episode. The choice of a neuroleptic lower in potency and dosage than that which precipitated the original NMS episode was not significantly related to successful outcome. PMID- 2569458 TI - Buspirone revisited. PMID- 2569459 TI - High-dose diazepam in neuroleptic-resistant schizophrenia with tardive dyskinesia. PMID- 2569460 TI - Induction of mitochondrial serine:pyruvate aminotransferase of rat liver by glucagon and insulin through different mechanisms. AB - Studies were performed in the rat liver to examine whether or not insulin as well as glucagon causes the induction of mitochondrial serine:pyruvate aminotransferase (SPTm) [EC 2.6.1.51] and if so, whether the mechanisms of induction are similar or different for the two hormones. Not only glucagon but also insulin induced SPTm. Cell-free translation assaying and RNA blot analysis showed that both hormones cause an increase in the hepatic level of mRNA for the precursor of SPTm. Their effects were virtually additive, and the time course of the increase in the mRNA level differed between the hormones. The maximal increase induced by glucagon was observed 3.5 h after the hormone injection while that by insulin was found after 6 h. The increase in the mRNA due to insulin was completely inhibited by the co-administration of cycloheximide, while that due to glucagon was not. The finding suggests that a newly synthesized, insulin dependent protein(s) is involved in the regulation of the mRNA level by insulin. On the other hand, hydrocortisone treatment selectively suppressed the increase in the mRNA due to glucagon. These data indicate that the synthesis of the mRNA for SPTm is regulated by glucagon and insulin through different mechanisms. The size of the hormone-induced mRNA for SPTm gradually decreased with time, but the cell-free translation products did not exhibit size alteration. RNase H digestion to remove the poly(A) tail of the mRNA indicated that shortening of the poly(A) sequence might be responsible for the time-dependent size alteration of the mRNA. PMID- 2569461 TI - Physical and chemical characterization of glutamine synthetase purified from Mycobacterium phlei. AB - Glutamine synthetase (L-glutamate: ammonia ligase [ADP forming]) [EC 6.3.1.2] has been purified from a Gram-positive, acid-fast bacterium, Mycobacterium phlei, by simple procedures with 57% recovery. The enzyme resembled that from Mycobacterium smegmatis in the subunit size (56,000), molecular weight (670,000), amino acid composition, the amino acid sequence of the NH2-terminal, and the secondary structure. The enzyme activity was regulated by adenylylation of each subunit in the dodecameric molecule. M. phlei glutamine synthetase possesses two useful characteristics: high thermostability and resistance to protease digestion. The enzyme was not inactivated on exposure to 60 degrees C for 2 h or 37 degrees C for 72 h, or after incubation with 1% trypsin or chymotrypsin at 37 degrees C for 12 h, pH 7.8. With saturating substrate levels, the Arrhenius plot was nonlinear and concave downward with an intersection point at 45 degrees C, and the activation energies were calculated to be 3.2 and 9.6 cal/mol from the slopes. The specific activity of the highly adenylylated enzyme (E10.7) was remarkably lower than that of the slightly adenylylated enzyme (E2.5); however, both enzymes show similar profiles of the Arrhenius plot. These results indicate that the adenylylation of the enzyme does not affect its activation energies. PMID- 2569462 TI - Genomic organization and chromosomal localization of the human cathepsin G gene. AB - Cathepsin G is a 26,000-Da serine protease that is found in the azurophil granules of neutrophils and monocytes. The cathepsin G gene is expressed at high levels in U937 promonocytic cells, but is down-regulated with phorbol-induced differentiation. To characterize the genomic sequences responsible for the regulated expression of this gene, we screened a human genomic fibroblast library using cathepsin G cDNA, and obtained two lambda clones that contained the cathepsin G locus. The cathepsin G gene spans 2.7 kilobase pairs of genomic DNA and consists of 5 exons and 4 introns. The genomic organization of cathepsin G is similar to that of human neutrophil elastase, rat mast cell protease II, murine adipsin, and murine cytotoxic T-cell serine proteases, with protease catalytic residues located near the borders of exons 2, 3, and 5. Using in situ hybridization techniques, we localized cathepsin G to chromosome 14q11.2, a site that is near the alpha/delta T-cell receptor complex. Cathepsin G transcription is abolished in U937 nuclei with 2 micrograms/ml alpha-amanitin, indicating that this gene is probably transcribed by RNA polymerase II. The 5' end of the cathepsin G gene was defined by primer extension and S1 nuclease protection assays. A TATA box is found at position -29, and a CAAT box is found at -69 with respect to the transcription initiation site. Having defined the genomic structure and chromosomal location of cathepsin G, we are now attempting to identify the DNA elements in or near this gene that mediate its tissue and development-specific pattern of expression. PMID- 2569463 TI - Post-translational suppression of expression of intestinal brush border enzymes by fructose. AB - The two major dietary sugars, fructose and sucrose, were found to suppress effectively the biosynthetic renewal of brush border enzymes in the gut. When studied in cultured explants of pig small intestine mucosa, 10-50 mM concentrations of fructose completely prevented the expression of mature aminopeptidase N and severely reduced that of sucrase-isomaltase. The instantly occurring and reversible suppressive effect manifested itself as a leupeptin sensitive degradation of newly synthesized brush border enzymes. The likely mechanism of action of the dietary sugar is by causing an abnormal cotranslational glycosylation that in turn triggers a rapid proteolytic breakdown. Our findings suggest that renewal of digestive brush border enzymes is transiently suppressed during intake of fructose- or sucrose-rich meals. PMID- 2569464 TI - Purification of a specific repressor of ferritin mRNA translation from rabbit liver. AB - The synthesis of ferritin is regulated at the translation level in coordination with iron availability. Under conditions of low iron, translation of ferritin mRNA is repressed and the majority of ferritin mRNA is non-polysomal. Upon an increase in iron, translation of ferritin mRNA is derepressed resulting in as much as a 50-100-fold increase in the rate of ferritin synthesis. This regulation is mediated at least in part by a specific translational repressor which binds to a conserved sequence, the iron responsive element, located in the 5'-untranslated region of ferritin mRNA. In this communication we report the purification of such a repressor from rabbit liver. This repressor, which we call the "ferritin repressor protein," has an apparent molecular mass of 90 kDa when analyzed by gel filtration chromatography. It inhibits translation of ferritin mRNA in a highly specific fashion when added to a wheat germ lysate programmed with liver poly(A+) mRNA. In addition, it binds specifically to sequences contained within the first 92 nucleotides of ferritin mRNA, most likely the iron responsive element. Analysis of highly purified repressor by sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows that it is composed primarily of a single polypeptide of approximately 90 kDa. Elution of this 90-kDa polypeptide from a sodium dodecyl sulfate gel followed by renaturation and analysis for repressor activity shows that it both binds to the 5'-untranslated region of ferritin mRNA and represses its translation in vitro. PMID- 2569465 TI - Regulation of proliferating cell nuclear antigen during the cell cycle. AB - The proliferating cell nuclear antigen (PCNA), also known as cyclin and DNA polymerase delta auxiliary factor, is present in reduced amounts in nongrowing cells and is synthesized at a greater rate in the S phase of growing cells. The recently discovered involvement of PCNA in DNA replication suggested that this pattern of expression functions to regulate DNA synthesis. We have investigated this possibility further by examining the synthesis, stability, and accumulation of PCNA in HeLa cells fractionated by centrifugal elutriation into nearly synchronous populations of cells at various positions in the cell cycle. In these fractionated cells we found that there is an increase in the rate of PCNA synthesis with a peak in early S phase of the cell cycle, but the magnitude of the increase is only 2-3-fold. This change reflects similar changes in the amount of PCNA mRNA. The fluctuating synthesis of PCNA maintains this protein at a roughly constant proportion of the total cell protein, although the amount doubles/cell in the cell cycle. Consistent with this observation, the stability of PCNA does not differ significantly from that of total cellular protein in synchronized HeLa cells. We also observed that a maximum of one-third of the total PCNA is tightly associated with the nucleus, presumably in replication complexes, at the peak of S phase. We conclude that the cyclic synthesis of PCNA in cycling HeLa cells maintains PCNA in excess of the amount involved directly in DNA replication and the amount of the protein neither fluctuates significantly with the cell cycle nor is limiting for DNA synthesis. PMID- 2569467 TI - Murine elongation factor 1 alpha (EF-1 alpha) is posttranslationally modified by novel amide-linked ethanolamine-phosphoglycerol moieties. Addition of ethanolamine-phosphoglycerol to specific glutamic acid residues on EF-1 alpha. AB - Elongation Factor 1 alpha (EF-1 alpha), an important eukaryotic translation factor, transports charged aminoacyl-tRNA from the cytosol to the ribosomes during poly-peptide synthesis. Metabolic radiolabeling with [3H] ethanolamine shows that, in all cells examined, EF-1 alpha is the major radiolabeled protein. Radiolabeled EF-1 alpha has an apparent Mr = 53,000 and a basic isoelectric point. It is cytosolic and does not contain N-linked oligosaccharides. Trypsin digestion of murine EF-1 alpha generated two major [3H]ethanolamine-labeled peptides. Three peptides were sequenced and were identical to two distinct regions of the human EF-1 alpha protein. Blank sequencing cycles coinciding with glutamic acid in the human cDNA-derived sequence were also found to release [3H]ethanolamine, and compositional analysis of these peptides confirmed the presence of glutamic acid. Dansylation analysis demonstrates that the amine group of the ethanolamine is blocked. These results indicate that EF-1 alpha is posttranslationally modified by the covalent attachment of ethanolamine via an amide bond to at least two specific glutamic acid residues (Glu-301 and Glu-374). The hydroxyl group of the attached ethanolamine was shown by mass spectrometry and compositional analysis, to be further modified by the addition of a phosphoglycerol unit. This novel posttranslational modification may represent an important alteration of EF-1 alpha, comparable to the regulatory effects of posttranslational methylation of EF-1 alpha lysine residues. PMID- 2569466 TI - Vitamin K-dependent carboxylation. In vitro modification of synthetic peptides containing the gamma-carboxylation recognition site. AB - Synthetic peptides including the gamma-carboxylation recognition site and acidic amino acids were compared as substrates for vitamin K-dependent gamma carboxylation by bovine liver carboxylase. The 28-residue proPT28 (proprothrombin -18 to +10) and proFIX28 (pro-Factor IX -18 to +10) were carboxylated with a Km of 3 microM. The Vmax of proPT28 was 2-3 times greater than that of proFIX28. An analog of proFIX28 that contained the prothrombin propeptide had a Vmax 2-3-fold greater than an analog of proPT28 that contained the Factor IX propeptide. proFIX28/RS-1, based upon Factor IX Cambridge, proFIX28/RQ-4, based upon Factor IX Oxford 3, and proFIX28 had equivalent Km and Vmax values. Analogs of proPT28 containing Ala6-Glu7 or Glu6-Ala7 were carboxylated at equivalent rates. A peptide containing Asp6-Asp7 was carboxylated at a rate of about 1% of that of Glu carboxylation. Carboxylation of peptides containing Asp6-Glu7 and Glu6-Asp7 yielded results identical with peptides containing Ala6-Glu7 and Glu6-Ala7. Carboxymethylcysteine was not carboxylated when substituted for Glu6 in a peptide containing Asp7. These results indicate that the prothrombin propeptide is more efficient in the carboxylation process than is the Factor IX propeptide, but that both propeptides direct carboxylation; the gamma-carboxylation recognition site does not include residues -4 and -1; aspartic acid and carboxymethylcysteine are poor substrates for the carboxylase, but aspartic acid does not inhibit the carboxylation of adjacent glutamic acids. PMID- 2569468 TI - Deletion and insertion mutants of the multidrug transporter. AB - The multidrug transporter is a 170,000-dalton membrane glycoprotein which confers multidrug resistance through its activity as an ATP-dependent efflux pump for hydrophobic, cytotoxic drugs. To determine the essential structural components of this complex membrane transporter we have altered an MDR1 cDNA in an expression vector by deletion and insertion mutations. The structure of the transporter deduced from its amino acid sequence suggests that it consists of two homologous, perhaps functionally autonomous, halves each with six transmembrane segments and a cytoplasmic ATP-binding domain. However, several carboxyl-terminal deletions, one involving 53 amino acids, the second removing 253 amino acids, and an internal deletion within the carboxyl-terminal half of the molecule, totally eliminate the ability of the mutant transporter to confer drug resistance. An internal deletion of the amino-terminal half, which removed residues 140-229, is also nonfunctional. Small carboxylterminal deletions of up to 23 amino acids leave a functional transporter, although the removal of 23 COOH-terminal amino acids reduces its ability to confer colchicine resistance. Insertions of 4 amino acids in a transmembrane domain, and in one of the two ATP-binding regions, have no effect on activity. These studies define some of the limits of allowable deletions and insertions in the MDR1 gene, and demonstrate the requirement for two intact halves of the molecule for a functional multidrug transporter. PMID- 2569470 TI - Gas-liquid chromatographic resolution of some racemic synthons for lamtidine analogous histamine H2-receptor antagonists via diastereomeric amides of (1S)-(-) camphanic acid. PMID- 2569469 TI - Acanthamoeba actin and profilin can be cross-linked between glutamic acid 364 of actin and lysine 115 of profilin. AB - Acanthamoeba profilin was cross-linked to actin via a zero-length isopeptide bond using carbodiimide. The covalently linked 1:1 complex was purified and treated with cyanogen bromide. This cleaves actin into small cyanogen bromide (CNBr) peptides and leaves the profilin intact owing to its lack of methionine. Profilin with one covalently attached actin CNBr peptide was purified by gel filtration followed by gel electrophoresis and electroblotting on polybase-coated glass fiber membranes. Since the NH2 terminus of profilin is blocked, Edman degradation gave only the sequence of the conjugated actin CNBr fragment beginning with Trp 356. The profilin-actin CNBr peptide conjugate was digested further with trypsin and the cross-linked peptide identified by comparison with the tryptic peptide pattern obtained from carbodiimide-treated profilin. Amino-acid sequence analysis of the cross-linked tryptic peptides produced two residues at each cycle. Their order corresponds to actin starting at Trp-356 and profilin starting at Ala-94. From the absence of the phenylthiohydantoin-amino acid residues in specific cycles, we conclude that actin Glu-364 is linked to Lys-115 in profilin. Experiments with the isoforms of profilin I and profilin II gave identical results. The cross-linked region in profilin is homologous with sequences in the larger actin filament capping proteins fragmin and gelsolin. PMID- 2569472 TI - Clonal loss of one chromosome 11 in a parathyroid adenoma. AB - Traditional cytogenetic approaches have been unsuccessful in the study of parathyroid adenomas. We now describe one parathyroid adenoma in which a molecular cytogenetic approach revealed clonal loss of one chromosome 11. Restriction fragment length polymorphism analysis of the patient's normal leukocyte DNA demonstrated heterozygosity at four loci (PTH, INT2, APOA1, and ETS1) that span the length of chromosome 11. However, the adenoma DNA showed clonal deletion of one allele, i.e. loss of heterozygosity, at each locus. Use of five nonpolymorphic probes from chromosome 11 was consistent with 50% loss of total chromosome 11 DNA in the adenoma. No tumor-specific loss of heterozygosity was observed when restriction fragment length polymorphisms from five other autosomes (no. 1, 5, 6, 7, and 12) were analyzed, and an X-chromosome probe also showed no tumor DNA loss. We have demonstrated tumor-specific chromosome loss in a parathyroid adenoma; such a lesion has been described only rarely in benign tumors. Our finding adds to the evidence for monoclonality in parathyroid adenomatosis, indicates that only one PTH gene copy is sufficient for hyperparathyroid tumor function, and raises the possibility that a tumor suppressor gene important in the development of nonfamilial parathyroid neoplasia is located on chromosome 11. PMID- 2569471 TI - Post-mortem immunodiagnosis of scrapie and bovine spongiform encephalopathy. AB - Two polyclonal antisera were raised in rabbits against the scrapie-associated fibril protein (PrP) prepared from sheep and mice which were terminally infected with experimental scrapie. The anti-mouse PrP serum identifies the proteins of scrapie-associated fibrils (SAF) from all the host species studied (mouse, hamster, sheep and goat) and bovine spongiform encephalopathy (BSE) fibrils from cow. The anti-sheep PrP serum displays species restricted immunoreactivity. While it identifies several PrP polypeptides from terminally affected sheep, goat and cow material, only the highest molecular weight band is recognised from hamster and there is no detection of mouse PrP. The use of these antisera in routine laboratory testing at post mortem provides a highly sensitive test for scrapie and BSE and may allow the identification of infected animals prior to the onset of clinical signs. PMID- 2569473 TI - Elevation of plasma epinephrine concentrations inhibits proteolysis and leucine oxidation in man via beta-adrenergic mechanisms. AB - The role of elevated plasma epinephrine concentrations in the regulation of plasma leucine kinetics and the contribution of beta-receptors were assessed in man. Epinephrine (50 ng/kg per min) was infused either alone or combined with propranolol (beta-blockade) into groups of six subjects fasted overnight; leucine flux, oxidation, and net plasma leucine forearm balance were determined during 180 min. Constant plasma insulin and glucagon concentrations were maintained in all studies by infusing somatostatin combined with insulin and glucagon replacements. Plasma leucine concentrations decreased from baseline during epinephrine infusion by 27 +/- 5 mumol/liter (P less than 0.02) due to a 22 +/- 6% decrease in leucine flux (P less than 0.05 vs. controls receiving saline) and to an increase in the metabolic clearance rate of leucine (P less than 0.02). Leucine oxidation decreased by 36 +/- 8% (P less than 0.01 vs. controls). beta Blockade abolished the effect of epinephrine on leucine flux and oxidation. Net forearm release of leucine increased during epinephrine (P less than 0.01), suggesting increased muscle proteolysis; the fall of total body leucine flux was therefore due to diminished proteolysis in nonmuscle tissues, such as splanchnic organs. Nonoxidative leucine disappearance as a parameter of protein synthesis was not significantly influenced by epinephrine. Plasma glucose and FFA concentrations increased via beta-adrenergic mechanisms (P less than 0.001). The results suggest that elevation of plasma epinephrine concentrations similar to those observed in severe stress results in redistribution of body proteins and exerts a whole body protein-sparing effect; this may counteract catabolic effects of other hormones during severe stress. PMID- 2569474 TI - Binding and internalization of somatostatin, insulin, and glucagon by cultured rat islet cells. AB - The pathways by which islet B, A, and D cells bind and internalize homologous (self) and heterologous (other) islet hormones were compared. [125I Tyr]Somatostatin-14 (S-14), 125I-insulin, and 125I-glucagon were incubated with monolayer cultures of neonatal rat islet cells. Tissues were processed for quantitative electron microscopic autoradiography by the probability circle method coupled to morphometry. For all three radioligands and all three cell types surface labeling was rapidly followed by internalization of the radioligands into endocytotic vesicles. The further intracellular movement of the ligand occurred in a time- and temperature-related manner and depended on whether it was homologous or heterologous for the cell in question. Thus [125I-Tyr]S-14 in B and A cells, 125I-insulin in A and D cells, and 125I-glucagon in B and D cells were rapidly transferred from endocytotic vesicles to lysosomal structures. By contrast, [125I-Tyr]S-14 in D cells, 125I-insulin in B cells, and 125I glucagon in A cells showed poor progression from endocytotic vesicles to downstream vesicular structures. We conclude that (a) each of the three radioligands is internalized by islet cells in a time- and temperature-dependent manner; (b) after initial internalization the further intracellular progression of the endocytosed radioligand occurs freely in cells heterologous for the radioligand but poorly in cells homologous for the radioligand; and (c) binding and endocytosis can be uncoupled from lysosomal degradation of ligand. PMID- 2569476 TI - A highly informative probe for two polymorphic Vh gene regions that contain one or more autoantibody-associated Vh genes. AB - Efforts to determine the role of specific Ig variable region (V) genes in human autoimmune responses have been hampered by the lack of suitably polymorphic probes. Recently we isolated a heavy chain V (Vh) gene, designated Humhv3005, that is 99% homologous to the 1.9III Vh gene and can encode an anti-DNA antibody. To study the relation between these two genes, different DNA fragments from the isolated Humhv3005 clone were used to probe Southern blots of human genomic DNA. A 1.6-kb Eco RI fragment (designated hv3005/E1.6) was found to hybridize with only one band in Eco RI-digested DNA, and with two major bands in Bam HI-digested DNA. Importantly, the sizes of the latter two bands were indistinguishable from the corresponding Bam HI fragment sizes of the isolated hv3005 clone and the isolated 1.9III clone, respectively. Population and family studies with the hv3005/E1.6 probe revealed five different hybridization patterns of these two characteristic bands, which defined nine possible genotypes for two human Ig Vh gene loci. Together the data demonstrate that hv3005/E1.6 is a highly informative probe for an autoantibody-associated Vh gene(s), and should prove useful in elucidating the role of Ig Vh genes in autoimmune diseases. PMID- 2569475 TI - Relaxation of rat vascular muscle by peripheral benzodiazepine modulators. AB - Effects of peripheral benzodiazepine receptor modulating drugs, Ro 5-4864 and PK 11195, on tension induced by K+ and the calcium agonist SDZ 202 791 (S isomer), were studied in rat caudal arteries. A significant reduction of tonic phase tension occurred with 30 nM PK 11195 or 3 microM Ro 5-4864, but decreases of the initial (first 3 min), phasic contraction were detected only at the highest concentrations of Ro 5-4864 and PK 11195. Protoporphyrin IX, the putative endogenous ligand of the peripheral benzodiazepine receptor, (at 10-100 nM) markedly increased the effectiveness of Ro 5-4864 and PK 11195 in reducing phasic contraction. Intracellular calcium localization and distribution in fura-2 loaded single vascular cells were quantitated using a high sensitivity, two-stage microchannel plate, photon-counting (PMI-VIM) camera. Peripheral benzodiazepines reduced intracellular calcium release from centrally located calcium pools, and this decrease of calcium release was potentiated by protoporphyrin IX. The decrease in intracellular calcium activity, which was more pronounced in the central regions where sarcoplasmic reticular elements are numerous, was probably the major mechanism of these vasodilator properties. Measurements of soluble guanylate cyclase activity also supported the intracellular Ca2+ release mechanism. Under conditions where protoporphyrin IX did not significantly stimulate guanylate cyclase, Ro 5-4864 alone or more effectively in combination with protoporphyrin IX stimulated cGMP production and caused relaxation. Guanylate cyclase forms a possible target for these benzodiazepine modulators, a hypothesis that merits further investigation. PMID- 2569477 TI - Clustering of multiallele DNA markers near the Huntington's disease gene. AB - Five highly informative multiallele restriction fragment length polymorphisms (RFLPs) of value for preclinical diagnosis of Huntington's disease (HD) have been genetically characterized. One RFLP was uncovered by expansion of the D4S43 locus while three others are at D4S111 and D4S115, loci defined by NotI-linking clones. The final marker, D4S125, represents a recently discovered VNTR locus. All four loci map closer to the HD gene and to the telomere than D4S10, the original linked marker for HD. In combination with two multiallele RFLPs previously identified for D4S43 and another linked locus, D4S95, these five new multiallele markers will dramatically improve the speed and accuracy of predictive testing in HD, and increase its applicability by maximizing the chances of an informative test for anyone with appropriate family structure. PMID- 2569478 TI - Hemopoietic origin of factor XIII A subunits in platelets, monocytes, and plasma. Evidence from bone marrow transplantation studies. AB - Factor XIII A subunit (FXIIIA) is found in plasma, platelets, and monocytes. The hemopoietic contributions to FXIIIA in these components were studied in patients transplanted with marrows from donors with different FXIIIA phenotypes. In three patients with successful engraftment (by DNA genotyping, red cell phenotyping, and cytogenetic studies) platelet and monocyte FXIIIA changed to donor phenotypes with hematologic recovery. Thus, FXIIIA in platelets and monocytes is synthesized de novo and/or from their progenitor cells. Plasma FXIIIA phenotype change after transplantation was more complex. Patient I changed from phenotype 1-1 (one electrophoretically fast band) to 1-2 (three bands) in 115 d; patients 2 and 3 did not change completely from phenotype 1-2 to 1-1 in up to 458 d, but did show enrichment of the fastest band. Thus, while there is a definite contribution of donor hemopoiesis to plasma FXIIIA, another source of recipient FXIIIA appears to be present to delay or prevent the phenotype change. PMID- 2569479 TI - Role of catalytic and lysine-binding sites in plasmin-induced neutrophil adherence to endothelium. AB - Plasmin resulted in increased neutrophil adherence to cultured ovine pulmonary artery endothelial cell monolayers in a concentration-dependent manner (10(-12) 10(-7) M). The adherence response increased fivefold above baseline within 60 min after addition of plasmin (10(-8) M) and the response persisted up to 30 min after removal of plasmin. The neutrophil adherence was mediated by the action of plasmin on neutrophils rather than endothelial cells. The response was the result of an increase in functional activity of CD18 neutrophil cell surface adhesive glycoprotein. Neutrophil adherence was inhibited by pretreatment of neutrophils with MAbs IB4 and 60.3 targeted against the beta chain of the CD18, whereas control isotypic MAb 60.5 against HLA class I antigen had no effect. The plasmin catalytic site was not involved in the response. Lys-plasminogen had reduced adherence-promoting activity relative to plasmin, whereas glu-plasminogen had no effect. Elastase-derived plasminogen fragments corresponding to kringle 1+2+3 and kringle 4 (both of which contained the lysine-binding sites) possessed neutrophil adherence-promoting activities similar to plasmin, whereas miniplasminogen (which contains the catalytic site but no lysine-binding sites) had minimal effect, indicating the involvement of lysine-binding sites in the response. Blocking lysine-binding sites of plasmin and elastase-derived plasminogen fragments with tranexamic acid (IC50 of 5 mM) inhibited neutrophil adherence. A monospecific polyclonal antibody against the lysine-binding sites also reduced the neutrophil adherence-promoting activity of plasmin. The results indicate that plasmin induces neutrophil adherence to the endothelium and that the effect is mediated by lysine-binding sites on plasmin. PMID- 2569480 TI - Coregulation of calcium channels and beta-adrenergic receptors in cultured chick embryo ventricular cells. AB - To examine mechanisms whereby the abundance of functional Ca channels may be regulated in excitable tissue, Ca channel number was estimated by binding of the dihydropyridine (DHP) antagonist 3H (+)PN200-110 to monolayers of intact myocytes from chick embryo ventricle. Beta adrenergic receptor properties were studied in cultured myocytes using [3H]CGP12177, an antagonist ligand. Physiological correlates for alterations in DHP binding site number included 45Ca uptake and contractile response to (+)BAYk 8644, a specific L-type Ca channel activator. All binding and physiological determinations were performed in similar intact cell preparations under identical conditions. 4-h exposure to 1 microM isoproterenol reduced cell surface beta-adrenergic receptor number from 44 +/- 3 to 17 +/- 2 fmol/mg (P less than 0.05); DHP binding sites declined in number from 113 +/- 25 to 73 +/- 30 fmol/mg (P less than 0.03). When protein kinase A was activated by a non-receptor-dependent mechanism, DHP binding declined similarly to 68% of control. Exposure to diltiazem, a Ca channel antagonist, for 18-24 h had no effect on number of DHP binding sites. After 4-h isoproterenol exposure, 45Ca uptake stimulated by BAYk 8644 declined from 3.3 +/- 0.2 nmol/mg to 2.9 +/- 0.3 nmol/mg (P less than 0.01) and BAYk 8644-stimulated increase in amplitude of contraction declined from 168 +/- 7 to 134 +/- 11% (P = 0.02). Thus, elevation of [cAMP] in myocytes is associated with a time-dependent decline in Ca channel abundance as estimated by DHP binding and a decline in physiological responses that are in part dependent on abundance of Ca channels. Binding of a directly acting Ca channel antagonist for 18-24 h does not modulate the number of DHP binding sites. PMID- 2569481 TI - A physiologic role for somatostatin 28 as a regulator of insulin secretion. AB - Somatostatin 28 (S-28) is a peptide produced in the intestinal tract which rises in the circulation during nutrient absorption. We tested the hypothesis that S-28 regulates B-cell function by (a) studying the effects on insulin secretion of "physiologic" infusions of S-28 and (b) measuring insulin responses during elevated nutrient-stimulated endogenous S-28 levels. (a) Synthetic S-28 was infused on separate days into six healthy men at rates of 25 and 50 ng/kg per h which mimicked postprandial levels. Subjects were given a bolus of glucose (0.1 g/kg) after 120 min. Insulin responses during S-28 infusions were compared to a control study using a saline infusion in the same individuals. Glucose-stimulated insulin secretion was inhibited during the infusion of 50 ng/kg per h S-28 when compared to control (P less than 0.05). (b) Insulin secretion during elevations of endogenous S-28 was studied in healthy men who received a bolus of 2.5 g arginine (n = 14) or 25 U of secretin (n = 8) 120 min after swallowing 50 g fat, or, on a separate day, an equivalent volume of water. S-28 levels rose significantly after fat ingestion but did not change after water. Arginine and secretin-stimulated insulin secretion was inhibited following ingestion of fat compared with intake of water (P less than 0.05). Arginine-enhanced glucagon secretion was not changed by fat ingestion. We conclude that elevations in plasma S-28 levels, occurring during the postprandial state, attenuate B-cell secretion and this peptide may be a physiologic modulator of nutrient-stimulated insulin release. PMID- 2569483 TI - Familial bone marrow monosomy 7. Evidence that the predisposing locus is not on the long arm of chromosome 7. AB - Loss of expression of a tumor-suppressing gene is an attractive model to explain the cytogenetic and epidemiologic features of cases of myelodysplasia and acute myelogenous leukemia (AML) associated with bone marrow monosomy 7 or partial deletion of the long arm (7q-). We used probes from within the breakpoint region on 7q-chromosomes (7q22-34) that detect restriction fragment length polymorphisms (RFLPs) to investigate three families in which two siblings developed myelodysplasia with monosomy 7. In the first family, probes from the proximal part of this region identified DNA derived from the same maternal chromosome in both leukemias. The RFLPs in these siblings diverged at the more distal J3.11 marker due to a mitotic recombination in one patient, a result that suggested a critical region on 7q proximal to probe J3.11. Detailed RFLP mapping of the implicated region was then performed in two additional unrelated pairs of affected siblings. In these families, DNA derived from different parental chromosome 7s was retained in the leukemic bone marrows of the siblings. We conclude that the familial predisposition to myelodysplasia is not located within a consistently deleted segment on the long arm of chromosome 7. These data provide evidence implicating multiple genetic events in the pathogenesis of myelodysplasia seen in association with bone marrow monosomy 7 or 7q-. PMID- 2569482 TI - Two common low density lipoprotein receptor gene mutations cause familial hypercholesterolemia in Afrikaners. AB - Familial hypercholesterolemia (FH), an autosomal dominant disease caused by mutations in the LDL receptor gene, is five times more frequent in the Afrikaner population of South Africa than it is in the population of the United States and Europe. It has been proposed that the high frequency is due to a founder effect. In this paper, we characterized 24 mutant LDL receptor alleles from 12 Afrikaner individuals homozygous for FH. We identified two mutations that together makeup greater than 95% of the mutant LDL receptor genes represented in our sample. Both mutations were basepair substitutions that result in single-amino acid changes. Each mutation can be detected readily with the polymerase chain reaction and restriction analysis. The finding of two common LDL receptor mutations in the Afrikaner FH homozygotes predicts that these mutations will predominate in the Afrikaner population and that the high frequency of FH is due to a founder effect. The increased incidence of ischemic heart disease in the Afrikaner population may in part be due to the high frequency of these two mutations in the LDL receptor gene. PMID- 2569484 TI - Comparison of western blot analysis and immunocytochemical detection of P glycoprotein in multidrug resistant cells. AB - A sensitive immunocytochemical technique was developed to detect a 170,000 dalton cell membrane glycoprotein (P-gp) in cell lines resistant to vincristine and vinblastine with varying degrees of resistance. P-gp was shown very clearly using the C219 monoclonal antibody and immunocytochemical detection with either antialkaline phosphate or peroxidase-antiperoxidase with silver gold intensification. There was good correlation between the results obtained with immunocytochemical detection of P-gp in single cells and Western blot analysis. The technique is easily performed and can detect P-gp in relatively small numbers of cells that Western blot analysis could miss and is suitable for clinical application. PMID- 2569485 TI - Augmentation of acetaminophen analgesia by the antihistamine phenyltoloxamine. AB - A double-blind, placebo-controlled, parallel-group study was performed to compare the analgesic activity of the combination of 650 mg acetaminophen plus 60 mg phenyltoloxamine citrate with that of 650 mg acetaminophen alone. Two hundred female inpatients who had severe pain associated with a recent episiotomy procedure were randomly assigned to receive a single dose of one of the two active treatments or a placebo. Analgesia was assessed over a 6-hour period. Treatments were compared on the basis of standard subjective scales for pain intensity and relief, a number of derived variables based on these data and two global measures. For essentially all measures, the two active treatments were significantly superior to the placebo control. The combination was significantly superior to acetaminophen alone for all analgesic measures including SPID, TOTAL, and global ratings. The results of this study demonstrate that 60 mg phenyltoloxamine produces significant augmentation of the analgesic activity of 650 mg acetaminophen in postepisiotomy pain. PMID- 2569486 TI - Impaired bioavailability of famotidine given concurrently with a potent antacid. AB - The effect of a high potency antacid on the oral bioavailability of a single dose of famotidine (40 mg) was evaluated in normal volunteers according to a randomized cross-over design. Ingestion of the antacid concurrently with famotidine resulted in a significant reduction of peak plasma famotidine concentration (from 156 +/- 22 to 104 +/- 7, P less than 0.05) and area under the famotidine plasma concentration curve (from 956 +/- 125 to 607 +/- 56, P less than 0.02). No significant interaction was observed when the antacid was ingested 2 hours after famotidine administration. PMID- 2569487 TI - Differential steroid hormone and neural influences on peptide mRNA levels in CRH cells of the paraventricular nucleus: a hybridization histochemical study in the rat. AB - The three major classes of neurons in the paraventricular nucleus (PVH) provide a rich model for studying hormonal and neural influences on multiple neuropeptides expressed in individual cells. A great deal of previous work has examined this problem at the immunohistochemical level, where hormonal and neural influences on peptide levels have been established. In situ hybridization methods were used here to determine whether these effects are accompanied by measurable changes in neuropeptide mRNA levels. In the first series of experiments, the time-course of corticosterone replacement effects on corticotropin-releasing hormone (CRH) mRNA levels in parvicellular neuroendocrine cells of adrenalectomized animals were determined, and a dose-response curve was established. CRH mRNA hybridization remains maximal with plasma levels of steroid up to about 50 ng/ml, then declines sharply between about 60-130 ng/ml, and is just detectable at higher levels. We confirmed that corticosterone decreases vasopressin mRNA levels in this cell group and showed that levels of preproenkephalin mRNA are also decreased, whereas no significant changes in cholecystokinin, beta-preprotachykinin, and angiotensinogen mRNA levels could be detected. Thus, corticosterone decreases some neuropeptide mRNA levels and has no influence on others in this cell group. Tyrosine hydroxylase mRNA hybridization is also unaffected in this part of the nucleus. In a second group of experiments, the cell-type specificity of corticosterone influences was examined. It was found that while the hormone depresses CRH mRNA levels in parvicellular neurons, it increases such levels in PVH neurons with descending projections, in certain magnocellular neurosecretory neurons, and in a part of the central nucleus of the amygdala, whereas no influence was detected in the rostral lateral hypothalamic area. Furthermore, the stimulatory effects of corticosterone have different threshold levels in different cell groups. Thus, in different types of neurons, corticosterone may increase, decrease, or have no influence on CRH mRNA levels. In contrast, while corticosterone depresses vasopressin mRNA levels in parvicellular CRH neurons, it has no obvious effects on vasopressin mRNA levels in magnocellular or descending neurons; as with CRH, the effects of corticosterone on vasopressin mRNA levels are cell-type specific. In a third series of experiments it was shown that glucocorticoid receptor and mineralocorticoid receptor mRNAs are found in all three cell types in the PVH and that corticosterone tends to produce modest increases in mRNA levels for both receptors. Finally, it was shown that unilateral catecholamine-depleting knife cuts do not change mRNA levels for any of the neuropeptides (or steroid hormone receptors) examined here, although dramatic changes in neuropeptide levels themselves have been shown.4+ PMID- 2569488 TI - A comparison of metronidazole and sulfasalazine in the maintenance of remission in patients with ulcerative colitis. AB - In a double-blind, randomized trial, we tested the effectiveness of metronidazole (0.6 g/day) against sulfasalazine (2 g/day) in the maintenance of remission in patients with ulcerative colitis. The patients were in remission for 1-11 months at entry to trial, which lasted for 12 months. Forty patients entered the trial and 33 completed it. Metronidazole was found to be slightly more effective than sulfasalazine, a difference statistically significant only at 12 months. Six patients also completed a crossover trial. Remission was maintained for 12 months in 3 patients by metronidazole and in none of the 6 by sulfasalazine. No significant side effects were noted, and in particular, no paresthesias were reported. This trial, as well as our previous one, suggests that metronidazole may be useful in the maintenance of remission in patients with ulcerative colitis, but that it is ineffective in the therapy of the acute attack. PMID- 2569489 TI - Management of bladder fistulas in Crohn's disease. AB - We reviewed the course of 500 patients with Crohn's disease to document the incidence, the nature, and the results of management of fistulas to the bladder. Seventeen patients (14 men and three women) had developed enterovesical fistulas: 16 had pneumaturia. The barium radiographs demonstrated the fistula in only 37%. All had received sulfasalazine, and most were treated with corticosteroids and antibiotics intermittently; two had successful control of their urinary symptoms on this regimen. Eight patients who received 6-mercaptopurine (6-MP) in addition tolerated the urinary fistula well, so that we encourage a trial of 6-MP for this complication of Crohn's disease. Six patients continue on medical therapy alone after a mean of 5.3 years. There were no instances of pyelonephritis during 60 patient years. Eleven patients eventually underwent bowel resection, but in only two was persistence of the enterovesical fistula the primary indication for elective surgery, and in both, it was the patient's choice. Visualization of the fistula on barium enema radiograph or presence of a connection between the sigmoid and the bladder were not associated with adverse outcome. An enterovesical fistula in Crohn's disease rarely leads to serious complications and can often be treated successfully with medical therapy alone: by itself, it need not serve as an indication for surgery. PMID- 2569490 TI - Mortality in patients who have their antihypertensive therapy changed. AB - Medical records were examined for 1935 patients who presented sequentially to a hypertension clinic between 1971 and 1981. Patients were classified according to whether they were on a beta-blocker, methyldopa, a potassium-losing diuretic, or whether they had discontinued any of these treatments. Age-standardized mortality rates were calculated and the relative risks of stopping compared with non stopping were computed. Those stopping a beta-blocker had a significantly higher mortality in the following year than those who continued, both in men [relative risk (RR) = 5.91, 95% confidence interval (Cl) 2.78-12.56] and women (RR = 5.67, 95% Cl 1.75-18.41). Moreover, women also had a significantly higher mortality when stopping methyldopa, compared with those who continued on the drug (RR = 4.91, 95% Cl 1.82-13.20). However, analysis of data from the years following withdrawal indicated that a high RR was not limited to the first year after the withdrawal of beta-blockers, but was still apparent in the fourth year after stopping. This indicates that the high mortality was not an early function of withdrawal. The high initial RR of mortality in women stopping methyldopa was followed by a substantial decrease in risk over the later years of follow-up. The high mortality in patients stopping particular antihypertensive drugs was not explained by known cardiovascular risk factors. PMID- 2569491 TI - A role for isotype-specific binding factors in the regulation of IgA- and IgG specific responses by the anti/contrasuppressor T cell circuit. AB - Previous studies have shown that the isotype of an antibody response is selected, in part, by the inhibition of isotype-specific suppression. The antisuppressor model predicts that isotype selection is initiated through an interaction between Ag, Ig, and a T cell-derived factor within 6 h of immunization. This report characterizes some of these molecules and their contribution to isotype regulation. Cultures of murine spleen cells stimulated with the T cell-dependent Ag SRBC led to Ag-specific IgG and IgA responses that could be suppressed and then antisuppressed by a molecular complex produced by mixing purified serum Ig with the supernatant of Ag-pulsed macrophages co-cultured with T cells. The supernatants from separate cultures of Ag-pulsed macrophages and rIL-1 alpha stimulated CD4+ T cells, could be pooled and mixed with Ig to produce functional antisuppressive complexes thereby allowing the factors from the different cell types to be studied separately. Adsorption of the co-culture or the rIL-1 alpha stimulated T cell supernatants against monoclonal IgG or IgA, removed IgG and IgA binding factors, respectively, and abrogated the ability to enhance the corresponding isotype. The adherent material could be recovered and used to reconstitute enhancement by the supernatants depleted of the binding factors. When affinity purified IgG or IgA was used as the source of Ig within the antisuppressive complexes, the enhancement of the antibody response was limited to the isotype of the regulatory Ig used to form the complex. Thus, manipulation of the antisuppressive molecules has a predictable effect on isotype selection. Release of isotype-specific binding factors by CD4+ cells by rIL-1 alpha supports the hypothesis that T cell circuits play a role in initiating isotype regulation. PMID- 2569492 TI - Determination of the components of immune complexes made in vitro with antigens derived from Pseudomonas aeruginosa. AB - A method for identification of the components of immune complexes would increase our understanding of the pathogenesis of chronic diseases such as Pseudomonas aeruginosa lung infection in cystic fibrosis. Capillary tube, gel diffusion and turbidimetric methods of determining immune complex formation were investigated with antigens from P. aeruginosa and the homologous rabbit antisera. Visible complexes were formed in the first two methods with flagella antigens. Purified lipopolysaccharide from P. aeruginosa would not form visible precipitates and a rapid and economical turbidimetric method was developed with 96-well microtiter plates. Larger quantities of immune complexes were formed in vitro with antigen/antibody ratios determined by the above methods. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting (Western blotting) were evaluated and found to be useful in determining the antigen and antibody components of these immune complexes. PMID- 2569493 TI - The distribution and duration of hantaan virus in the body fluids of patients with hemorrhagic fever with renal syndrome. AB - The distribution and duration of Hantaan virus (HTNV) in the body fluids of patients were studied by immunofluorescence, reverse passive hemagglutination, and cell culture assays. Virus antigen of hemorrhagic fever with renal syndrome in peripheral blood mononuclear cells (PBMCs) was usually present before day 11 of the disease, especially from days 4-7. Virus isolates were more readily recovered from plasma early in the course of the illness and less frequently after day 7. The use of PBMCs rather than plasma enabled isolates to be recovered at a rate nearly twice that permitted by plasma and allowed the isolation peak of HTNV (days 4-7 after onset of disease) to extend an additional 2 or 3 d, thus prolonging the period of detectable viremia until days 8-11. PBMCs were especially useful in isolating viruses from patients with hemorrhagic fever with renal syndrome in whom antibody titers were generally high during the acute phase of the disease. HTNV was isolated from the cerebrospinal fluid of patients, but was difficult to recover from other body fluids. PMID- 2569494 TI - Lectin-mediated, nonopsonic phagocytosis of type 1 Escherichia coli by human peritoneal macrophages of uremic patients treated by peritoneal dialysis. AB - Human peritoneal macrophages isolated from uremic patients undergoing peritoneal dialysis bind type 1 fimbriated Escherichia coli in the absence of opsonins. The number of bacteria bound per macrophage was 6.9, as determined by microscopic examination. Methyl alpha-mannoside (0.1 mM) and p-nitrophenyl alpha-mannoside (0.01 mM) inhibited this binding by about 66%. The ability of peritoneal macrophages to bind E. coli in a mannose-specific manner was confirmed in further experiments using an enzyme-linked immunosorbent assay (ELISA) with an antibacterial antibody, radiolabelled E. coli, and counts of colony-forming units (CFU). The number of bacteria bound per macrophage was 7 to 12 in the ELISA and 5.5-8.5 in the CFU assay. Methyl alpha-mannoside caused 70% inhibition of binding in the ELISA and 84% in the CFU assay, whereas p-nitrophenyl alpha-mannoside showed inhibition of 79% and 90%, respectively. Most bound bacteria (76-80%) were subsequently killed. Nonfimbriated E. coli 827 bound poorly to the macrophages (approximately 22%) as compared to that of the fimbriated bacteria. Although this binding was not inhibited by methyl alpha-D-mannoside or p-nitrophenyl alpha mannoside, the percentage of bacteria killed was similar to that of the fimbriated phenotype. The peritoneal macrophage is thus able to phagocytose E. coli in the absence of opsonins. This may explain the relative rarity of E. coli as an etiologic agent of peritoneal infections in the dialysed patient. PMID- 2569495 TI - Levels of tyrosinase and its mRNA in coat-color mutants of C57BL/10J congenic mice: effects of genic substitution at the agouti, brown, albino, dilute, and pink-eyed dilution loci. AB - The levels of tyrosinase activities, tyrosinase cross-reacting material (TY-CRM), and tyrosinase mRNA were determined for skins from five congenic strains of mice (Mus musculus) with different coat colors. The dopa oxidase activity and melanin formation activity were directly proportional to the abundance of TY-CRM in all of the mutants. The levels of the enzyme activities and TY-CRM were increased in brown (b/b) and dilute (d/d) mice, while they were reduced in agouti (A/A) and pink-eyed dilution (p/p) animals. In albino (c/c) mice, the tyrosinase activities and melanin formation activity were scarcely detectable, and no TY-CRM was observed. Contrary to these, the levels of tyrosinase mRNA in agouti and albino mutants were almost the same as that in black mice, and those in brown and dilution mutants were 20-30% higher than that in black mice. These results indicate that the mutations at the brown and dilute loci exert their influence on the tyrosinase mRNA level, presumably by affecting the transcription of the tyrosinase gene, and that the mutations at the agouti and albino loci exert their influence on the TY-CRM level by regulating translation of tyrosinase mRNA and/or post-translation modification of the enzyme. PMID- 2569496 TI - Beta-adrenoceptors in the extraorbital lacrimal gland of the Syrian hamster. Characterization with [125I]-iodopindolol and evidence of sexual dimorphism. AB - Saturation and competition experiments with the radiolabeled beta-adrenergic antagonist (--)-[125I]-iodopindolol were used to characterized beta-adrenoceptor density (Bmax) and receptor affinity in the extraorbital lacrimal gland of male and female Syrian hamsters. Specific binding to the receptor was saturable. Scatchard analysis of saturation isotherms revealed a single population of receptor sites. Male glands had a significantly higher Bmax (38.9 +/- 5.0 vs. 23.3 +/- 2.1 fmol/mg protein, means +/- SEM, p less than 0.02) and receptor affinity (expressed in a lower dissociation constant Kd: 0.065 +/- 0.013 vs. 0.120 +/- 0.015 nM, p less than 0.02) than female glands. Binding of the radiolabeled ligand in competition with various adrenergic antagonists showed the receptor to be stereospecific and of the beta 2-subtype. PMID- 2569497 TI - Antagonism by pindolol, but not betaxolol, of 8-OH-DPAT-induced facilitation of male rat sexual behavior. AB - 8-OH-DPAT (0.25 mg/kg s.c.) produced a facilitation of the male rat sexual behavior, characterized by a decrease in the number of intromissions preceding ejaculation and in the time to ejaculation. This facilitation of the sexual behavior was antagonized by administration of the 5-HT and beta-adrenoceptor antagonist pindolol (4 mg/kg i.p.), but not by the selective beta-adrenoceptor antagonist betaxolol (4 mg/kg i.p.). Neither pindolol (2-8 mg/kg), nor betaxolol (2-8 mg/kg), produced any statistically significant effects per se on the male rat sexual behavior, as observed here (mounts, intromissions, ejaculation latency or the post-ejaculatory interval). A higher dose (16 mg/kg) of betaxolol produced a statistically significant reduction in the number of intromissions preceding ejaculation and in the ejaculation latency. The antagonism by pindolol of 8-OH DPAT-induced effects on male rat sexual behavior suggests an involvement of 5 HT1A receptors in the facilitation of this behavior produced by 8-OH-DPAT. PMID- 2569498 TI - The partial dopamine receptor agonists (--)-3-PPP and transdihydrolisuride enhance prolactin secretion in female but not in male rats. AB - The effects exerted by the partial dopamine (DA) receptor agonists (--)-3-(3 hydroxyphenyl)-N-n-propylpiperidine [(--)-3-PPP] and transdihydrolisuride (TDHL) on prolactin secretion were compared in male and female rats. Both agents were found to increase prolactin release more than tenfold in female rats. In male rats, in contrast, (--)-3-PPP and TDHL were either without effect or decreased prolactin secretion. It is suggested that this sexually differentiated prolactin response to partial DA agonists reflects a lower responsiveness of lactotroph DA receptors in female than in male rats. PMID- 2569499 TI - Effects of cadmium on quantal transmitter release and ultrastructure of frog motor nerve endings. AB - Exposure of frog cutaneous pectoris nerve-muscle preparations to cadmium (0.1-1 mM) results in an increase in miniature end-plate potential (m.e.p.p.) frequency. The increase is dependent on the concentration, the time of exposure and the co presence of other divalent cations in the extracellular fluid. The stimulatory effect of cadmium is most marked in a calcium-free medium. Increased levels of calcium (4-10 mM) or of magnesium (10 mM) reduce the stimulatory effect suggesting that those cations interfere with the entry of cadmium into nerve endings. Once the effect of cadmium on m.e.p.p. frequency is attained, washing with a cadmium-free solution fails to abolish its effect. The action of cadmium on m.e.p.p. frequency slowly declines towards zero after about 3 hrs. An ultrastructural study of nerve terminals exposed for one hr to 1 mM cadmium reveals that neither in calcium-containing nor in a nominally calcium-free medium are there any significant changes in the number of synaptic vesicles as compared to controls. However, after 3 hrs of cadmium action in a calcium-free medium there is about 65% depletion of synaptic vesicles, while in calcium-containing media there is only about 25% depletion. The results suggest that cadmium by itself can support transmitter release but not synaptic vesicle recycling which instead might depend upon calcium. PMID- 2569500 TI - Reduced glycine stimulation of [3H]MK-801 binding in Alzheimer's disease. AB - The novel N-methyl-D-aspartate receptor channel ligand (+)-[3H]5-methyl-10,11 dihydro-5H-dibenzo[a,d]-cyclohepten-5, 10-imine maleate ([3H]MK-801) has been utilized to label this receptor in human brain tissue. Characteristics of [3H]MK 801 binding to well-washed membranes from 17 control subjects and 16 patients with Alzheimer's disease were determined in frontal, parietal, and temporal cerebral cortex and cerebellar cortex. In control tissue the pharmacological specificity of the binding of this substance is entirely consistent with the profile previously reported for rat brain. Binding could be stimulated by the addition of glutamic acid to the incubation medium; addition of glycine produced further enhancement which was not prevented by strychnine. The specificity of the effects of these and other amino acids on the binding was the same as in the rat. In Alzheimer's disease significantly less binding was observed in the frontal cortex under glutamate- and glycine-stimulated conditions. This appears to be associated with a reduced affinity of the site whereas the pharmacological specificity of the site remained unchanged. The effect did not appear to be due to differences in mode of death between Alzheimer's disease and control subjects and is unlikely to be related to factors for which the groups were matched. In contrast, binding was not altered in the absence of added amino acids and presence of glutamate alone. These results imply that in the cerebral cortex the agonist site and a site in the cation channel of the receptor are not selectively altered, but that their coupling to a strychnine-insensitive glycine recognition site is impaired. PMID- 2569501 TI - Glutamate transport and not glutamate receptor binding is stimulated by gangliosides in a Ca2+-dependent manner in rat brain synaptic plasma membranes. AB - Crude as well as purified synaptic plasma membrane (SPM) preparations were analyzed for the influence of the ganglioside galactosyl-N-acetylgalactosaminyl (N-acetylneuraminyl)-galactosylgluc osyl ceramide (GM1) on high-affinity binding of L-[3H]glutamate. Assayed in two different buffer systems, SPM consistently exhibited increased (40-50%) binding upon incubation with GM1 plus Ca2+, as compared to controls without GM1. Incorporation experiments with 3H-labeled GM1 proved trypsin-stable insertion of GM1 into SPM, with a maximum incorporation of four times the endogenous amount (35 nmol/mg of protein). The observed increase in glutamate binding was not due to a change in the affinity of the binding sites, but to a change in the number of binding sites, and it was absolutely dependent on the presence of Ca2+. A pharmacological profile of the GM1/Ca2+ stimulated glutamate binding is presented. The original classification of the stimulatory effect as an effect on glutamate receptor binding had to be revised to take into account the observed temperature sensitivity of the ganglioside effect, its sensitivity to high osmolarity and to ultrasonication, and the lack of binding stimulation after detergent treatment of membranes or after receptor solubilization. Vesicular space measured in both SPM preparations was found to be around 7 microliters/mg of protein, in ganglioside-treated as well as in control membranes. From the data, it is concluded that a special, Na+- and Cl- independent form of glutamate transport into resealed membrane vesicles is stimulated by gangliosides in the presence of Ca2+. PMID- 2569502 TI - Glycine potentiates the stimulation of inositol phospholipid hydrolysis by excitatory amino acids in primary cultures of cerebellar neurons. AB - Glycine potentiates stimulation of inositol phospholipid hydrolysis by glutamate and N-methyl-D-aspartate, but not by quisqualate or carbamylcholine, in primary cultures of cerebellar granule cells. This potentiation occurs in the absence of extracellular Mg2+, but is more evident when stimulation of inositol phospholipid hydrolysis by N-methyl-D-aspartate is measured in the presence of 1 mM Mg2+. The action of glycine is not antagonized by strychnine. These results suggest that glycine acts as a positive modulator of signal transduction at a specific class of N-methyl-D-aspartate-sensitive glutamate receptors coupled to inositol phospholipid hydrolysis in cerebellar granule cells. PMID- 2569503 TI - Synthesis of L-3,4-dihydroxyphenylalanine by tyrosine hydroxylase cDNA transfected C6 cells: application for intracerebral grafting. AB - In the present study, we obtained genetically manipulated nonneuronal cells which synthesize a catecholamine precursor for future use in intracerebral grafting. Human type 1 tyrosine hydroxylase (TH; EC 1.14.16.2) cDNA was inserted into eukaryotic expression vector pKCRH2 and was co-transfected into C6 cells with plasmid pSV2neo. Expression of the TH minigene was screened by immunohistochemical staining with TH antibody and immunoblotting analysis. Several clones of the C6 transfectants that produce TH molecules were obtained. These cells showed TH activity, and the product, L-3,4-dihydroxyphenylalanine (L DOPA), was detected intracellularly due to the absence of L-amino acid decarboxylase (EC 4.1.1.28) activity. It was found that a large amount of L-DOPA was released from the cells into the culture medium. These transfectants were transplanted into rat brain, and the expression of TH was examined immunohistochemically. On the 10th day following transplantation, a mass of C6 cells which was heavily stained with TH antibody was observed in the brain. These findings may provide us with an opportunity to investigate the effects of intracerebral transplantation of nonneuronal cells that produce catecholamine or its precursor. PMID- 2569504 TI - Regulation of sigma-receptors: high- and low-affinity agonist states, GTP shifts, and up-regulation by rimcazole and 1,3-Di(2-tolyl)guanidine. AB - The regulation of the central sigma-binding site was investigated using both in vitro and in vivo manipulations in conjunction with radioligand binding. The displacement of the binding of R(+)-[3H]3-[3-hydroxyphenyl]-N-(1 propyl)piperidine [R(+)-[3H]3-PPP] to cortical homogenates by a range of drugs was consistent with the site labelled being a sigma-receptor. (+)-SKF 10,047, (-) SKF 10,047, (+/-)-cyclazocine, phencyclidine, and dexoxadrol displaced R(+)-[3H]3 PPP with pseudo-Hill coefficients of less than 1. Further analysis employing nonlinear curve fitting techniques demonstrated that displacement data for these compounds were described better by a model whereby R(+)-[3H]3-PPP was displaced from two discrete sites; approximately 65% of the total sites were in the high affinity state. In the presence of 10 mM Mg2+ and 0.3 mM GTP, displacement curves for (+)-SKF 10,047 and (+/-)-cyclazocine were shifted to the right. These findings were due to the shift of some 15% of the high-affinity binding sites to a low-affinity state. Saturation experiments revealed that 0.3 mM GTP acted competitively to decrease the affinity of R(+)-[3H]3-PPP for the sigma sites. The sigma-binding site was thus likely to be linked to a guanine nucleotide regulatory (G) protein. Thus sigma drugs could be subdivided on the basis of their GTP sensitivity and pseudo-Hill coefficients, and by analogy with other receptors R(+)-3-PPP, (+)-SKF 10,047, and (+/-)-cyclazocine, may be putative sigma-agonists. 1,3-Di(2-tolyl)guanidine (DTG), rimcazole, and haloperidol displaced R(+)-[3H]3-PPP with pseudo-Hill coefficients of approximately unity and thus may be sigma-antagonists. Subchronic treatment with rimcazole was characterized by slight sedation and a concomitant up-regulation, with a decrease in the affinity, of sigma-binding sites. The schedule of rimcazole also increased dopamine turnover in the nucleus accumbens; both the concentration of 3,4 dihydroxyphenylacetic acid (DOPAC) and the DOPAC/dopamine ratio were elevated. DTG produced similar alterations to the binding parameters of the sigma-binding site; however, changes were not observed in general behavior or accumbal dopamine turnover. sigma-Receptors are likely to be linked to a G protein and are functionally involved in the CNS. PMID- 2569505 TI - Direct transfer into nitrocellulose and quantitative radioautographic anatomical determination of brain tyrosine hydroxylase protein concentration. AB - An improved quantitative immunochemical determination of brain tyrosine hydroxylase (TH) concentrations was designed using direct transfer into nitrocellulose from 20-microns thick brain sections, followed by immunodetection and quantitative radioautography in three reference brain structures (locus ceruleus, substantia nigra, and ventral tegmental area). Results obtained by this methodology were similar to those obtained after extraction and Western blotting of the TH protein in control and reserpine-treated animals. Moreover, this methodology allows the combination of high sensitivity and high anatomical resolution in the study of the distribution of pharmacological effects. The locus ceruleus exhibited a significant posteroanterior distribution of TH protein concentration in control and reserpine-treated animals. PMID- 2569506 TI - Regulation of cyclic AMP formation in cultures of human foetal astrocytes by beta 2-adrenergic and adenosine receptors. AB - Two cell cultures, NEP2 and NEM2, isolated from human foetal brain have been maintained through several passages and found to express some properties of astrocytes. Both cell cultures contain adenylate cyclase stimulated by catecholamines with a potency order of isoprenaline greater than adrenaline greater than salbutamol much greater than noradrenaline, which is consistent with the presence of beta 2-adrenergic receptors. This study reports that the beta 2 adrenergic-selective antagonist ICI 118,551 is approximately 1,000 times more potent at inhibiting isoprenaline stimulation of cyclic AMP (cAMP) formation in both NEP2 and NEM2 than the beta 1-adrenergic-selective antagonist practolol. This observation confirms the presence of beta 2-adrenergic receptors in these cell cultures. The formation of cAMP in NEP2 is also stimulated by 5'-(N ethylcarboxamido)adenosine (NECA) more potently than by either adenosine or N6-(L phenylisopropyl)adenosine (L-PIA), which suggests that this foetal astrocyte expresses adenosine A2 receptors. Furthermore, L-PIA and NECA inhibit isoprenaline stimulation of cAMP formation, a result suggesting the presence of adenosine A1 receptors on NEP2. The presence of A1 receptors is confirmed by the observation that the A1-selective antagonist 8-cyclopentyl-1,3-dipropylxanthine reverses the inhibition of isoprenaline stimulation of cAMP formation by L-PIA and NECA. Additional evidence that NEP2 expresses adenosine receptors linked to the adenylate cyclase-inhibitory GTP-binding protein is provided by the finding that pretreatment of these cells with pertussis toxin reverses the adenosine inhibition of cAMP formation stimulated by either isoprenaline or forskolin. PMID- 2569507 TI - Tyrosine hydroxylase content of residual striatal dopamine nerve terminals following 6-hydroxydopamine administration: a flow cytometric study. AB - Fluorescence-activated cell sorting based on immunolabeling with a monoclonal antibody to tyrosine hydroxylase and a fluorescein-conjugated secondary antibody was used to identify striatal synaptosomes derived from nigrostriatal dopamine nerve terminals. The amount of tyrosine hydroxylase immunoreactivity in dopaminergic striatal synaptosomes prepared from control rats was compared to the amount in dopaminergic synaptosomes prepared from rats that had received intraventricular injections of 6-hydroxydopamine. Although the absolute number of dopaminergic synaptosomes was decreased in lesioned animals, those residual dopamine terminals present contained more tyrosine hydroxylase than did dopamine terminals from control rats. Both the decrease in the absolute number of dopamine terminals and the increase in tyrosine hydroxylase immunoreactivity in residual terminals were proportional to the extent of the lesion, as determined by measurement of striatal dopamine levels. These results suggest that an increase in the amount of tyrosine hydroxylase protein in residual terminals may represent one compensatory mechanism by which residual dopamine neurons maintain normal striatal function after partial destruction of the nigrostriatal dopamine projection. PMID- 2569508 TI - Linkage analysis approach to hereditary movement disorders. PMID- 2569509 TI - Legal vulnerabilities in the treatment of chemically dependent dual diagnosis patients. AB - Substance abusing patients with significant psychiatric disorders can present unique legal vulnerabilities for treatment professionals. The tendency within the chemical dependency community to treat patients uniformly can militate against the individualized treatment planning that satisfies legal obligations to act in the best interest of the patient. The failure to adequately consider both the psychiatric disorder and substance abuse problem may create legal vulnerabilities that issue from the failure to refer a patient in need of medication or the failure to prevent the suicide of a patient in a situation where suicide is preventable. Patients suffering harm from acts of commission or omission of a chemical dependency treatment specialist have recourse through laws regarding negligence. Variations in the backgrounds and training of care providers create problems for the harmed patient in establishing a standard of care for the treatment of chemical dependence. However, the psychological symptoms of the dual diagnosis patient make it easier to establish a standard of care that may be legally cognizable. The chemical dependency specialist--of whatever discipline- has a duty to patients that requires sound assessment and disposition. PMID- 2569510 TI - A new approach to encapsulating nonsteroidal anti-inflammatory drugs. III. Coating acidic as well as basic nonsteroidal anti-inflammatory drugs with cellulose derivatives having different functional groups. AB - Acidic and basic nonsteroidal anti-inflammatory drugs were encapsulated with cellulose derivatives having different functional groups, acidic (carboxymethyl cellulose), basic (chitosan), and neutral (hydroxypropylmethyl cellulose). The properties of the microcapsules were studied and the interaction of the drugs with chitosan was assessed. PMID- 2569511 TI - A new approach to encapsulating nonsteroidal anti-inflammatory drugs. IV. Effect of cellulose derivatives with different functional groups on the bioavailability and gastric ulcerogenic activity of acidic as well as basic nonsteroidal anti inflammatory drugs. AB - The bioavailability and gastric ulcerogenic activity of oxyphenbutazone and glafenine (acidic and basic nonsteroidal anti-inflammatory drugs), coated with different cellulose derivatives were assessed in albino rats. The cellulose derivatives chosen have different functional groups, acidic (carboxymethyl cellulose), basic (chitosan) and neutral (hydroxypropylmethyl cellulose). The bioavailability was dependent on the drug and polymers. Generally, all the cellulose derivatives chosen decreased the gastric ulcerogenic activity of the drugs studied. PMID- 2569513 TI - The material tensile strength of convex-faced aspirin tablets. AB - The material tensile strength of a range of convex-faced tablets, compacted under controlled conditions at pressures of 40 and 320 MPa from a size fraction of acetylsalicylic acid, has been assessed. The calculation of the tensile strength sigma 1, from observed fracture loads obtained in diametral compression testing, is based on the equation derived by Pitt et al (1988), namely: (formula; see text) where P is the fracture load, D is the tablet diameter, t is the overall tablet thickness and W is the central cylinder thickness. The strength of a tablet of a given shape compacted at 320 MPa was between two and four times greater than that of a similar tablet compacted at 40 MPa. For the thicker tablets (W/D greater than or equal to 0.2) the material tensile strength was practically independent of shape. For the thinner tablets (W/D = 0.1) the material tensile strength varied considerably with face-curvature, showing a maximum for each of the two compaction pressures at a D/R value of 0.67. PMID- 2569512 TI - Alpha-blocking potencies of antihypertensive agents (prazosin, terazosin, bunazosin, SGB-1534 and ketanserin) having with quinazoline or quinazolinedione as assessed by radioligand binding assay methods in rat brain. AB - The comparative effects of antihypertensive agents, quinazoline or quinazolinedione residues (prazosin, bunazosin, terazosin, SGB-1534, and ketanserin), on the binding of [3H]prazosin, [3H]p-aminoclonidine and [3H]dihydroalprenolol([3H]DHA) to alpha 1-, alpha 2-, and beta-adrenoceptors in the rat brain were examined using radioligand binding assay methods. pA2 values were also obtained in the isolated rat aorta (alpha 1-adrenoceptor) using phenylephrine as an agonist. A strong inhibition by these drugs of [3H]prazosin binding to alpha 1-adrenoceptors was observed, while the inhibition of [3H]DHA binding to beta-adrenoceptors and [3H]p-aminoclonidine binding to alpha 2 adrenoceptor was found to be very weak. The rank order of antagonistic potencies of these drugs against the alpha 1-adrenergic receptors was determined by inhibition constants (Ki) with SGB-1534 = prazosin = bunazosin greater than terazosin greater than ketanserin. The pA2 value of these drugs, in contrast, had prazosin with higher pA2 value than that of SGB-1534. From these two different types of experiments, it was clear that these drugs antagonized alpha 1 adrenoceptors even in the central nervous system, and the side chains bound to quinazoline and quinazolinedione residues may play an important role in the antagonistic potencies for alpha 1-adrenoceptors in the central nervous system as in the peripheral tissues. PMID- 2569514 TI - Quantitative design for photostabilization of nifedipine by using titanium dioxide and/or tartrazine as colourants in model film coating systems. AB - The photostabilization of nifedipine by using film coating has been investigated in model systems in which a drug sample, dispersed on a glass plate, was covered with a free film and exposed to intensive light of mercury vapour lamp. The light transmission properties of films containing titanium dioxide or tartrazine alone as colourant were not always satisfactory. The tartrazine system exhibited superior light transmission properties to the titanium dioxide system at all additive concentrations. However, in combination both colourants gave much better light protection than did the colourants separately. The film-coated drug degraded following apparent first-order kinetics. The degradation rate constant decreased as both colourant concentration and film thickness increased; thus photostabilization was almost completely achieved by applying a film (thickness: 60 microns) of the binary mixture system containing only 0.7% of each of these colourants. The protective effectiveness of a film could be quantitatively evaluated by plotting the degradation rate constant against CL value of film formulation (C: concentration of colourant; L: film thickness). The degradation rate constant showed a good linear correlation with the average percent transmittance of a film in the wavelength range relating to the photolytic degradation of the drug. The average percent transmittance was thus proved to be an important and useful parameter for estimating the photostability of film coated drugs. PMID- 2569515 TI - The effect of hydrotropic agents on the heat coagulation of bovine serum albumin. AB - The effect of two hydrotrophic solubilizers on the heat coagulation of bovine serum albumin (BSA) has been investigated. Photon correlation spectroscopy indicated possible unfolding of BSA molecules in solutions of sodium benzoate and sodium salicylate at 25 degrees C. The effect of these hydrotropes on the heat coagulation of BSA was concentration-dependent. Relatively low concentrations stabilized the protein structure as indicated by the increase in the transition temperature(Tm) and induced gelation at temperatures and BSA concentrations lower than those required in the absence of hydrotropes. Higher concentrations of the hydrotropes considerably reduced Tm and inhibited gelation of BSA, the effect of sodium salicylate being more pronounced, as was the lower aggregation rate of BSA. The behaviour of these hydrotropes as protein denaturants differs from that of neutral electrolytes but is similar to that of concentrated solutions of urea. PMID- 2569516 TI - Effect of beta-casomorphins on intestinal propulsion in the guinea-pig colon. AB - beta-Casomorphins are a family of opioid peptides originally isolated from beta casein. In view of a possible physiological significance of these milk-derived compounds, the effects of bovine beta-casomorphin-5 (beta-CM-5), beta-casomorphin 4 (beta-CM-4) and D-Ala2-beta-casomorphin-4-NH2 (D-Ala2-beta-CM-4-NH2) have been investigated on the peristaltic reflex in the guinea-pig isolated colon and compared with morphine. beta-CM-5 and D-Ala2-beta-CM-4-NH2 each dose-dependently inhibited the velocity of propulsion of an intraluminal bolus; beta-CM-4 was ineffective. IC50 values were 0.30, 5.21 and 0.29 microM for morphine, beta-CM-5 and D-Ala2-beta-CM-4-NH2, respectively. The potency ratios vs morphine were 0.06 and 0.96 for beta-CM-5 and D-Ala2-beta-CM-4-NH2, respectively. Blockade of the peristaltic reflex by beta-CM-5 or D-Ala2-beta-CM-4-NH2 was reversed by the opioid antagonist naloxone. D-Ala2-beta-CM-4-NH2 also dose-dependently inhibited resting acetylcholine output (IC50 = 5.69 microM; potency ratio vs morphine: 0.63). In conclusion, certain beta-casomorphins inhibit intestinal propulsion and cholinergic neurotransmission in the guinea-pig colon, probably by acting at opioid receptors. PMID- 2569517 TI - Effects of trimebutine maleate on electrical activities of isolated mammalian cardiac preparations. AB - The effects of trimebutine maleate on electrical activity in guinea-pig isolated papillary muscles and rabbit sino-atrial nodes have been studied by means of a standard microelectrode method. In papillary muscles, trimebutine (above 10 microM) decreased the maximum rate of rise (Vmax) and the action potential duration at 90% repolarization (APD90), whereas the resting potential was not significantly altered. As to a decrease in Vmax, trimebutine produced a negative shift of the curve relating Vmax to the resting potential along the voltage axis. Trimebutine also depressed the slow action potentials of papillary muscles produced by 27 mM K and 0.2 mM Ba. In spontaneously beating sino-atrial node preparations, trimebutine (above 10 microM) decreased the heart rate, Vmax and the rate of diastolic depolarization. These results indicate that trimebutine maleate possesses a depressant action on the electrical activities of the fast- and slow-response fibres of the heart mainly due to inhibitions of both fast Na+ and slow Ca2+ channels. PMID- 2569518 TI - Non-specific hyperreactivity to pharmacological stimuli in tracheal and lung parenchymal strips of actively sensitized guinea-pigs. AB - The responsiveness of tracheal and lung parenchymal strips isolated from actively sensitized guinea-pigs to CaCl2 (in K+-depolarized tissue), KCl, acetylcholine and histamine was compared with that of strips from unsensitized animals. The concentration-response curves to the mentioned agonists exhibited, in the sensitized group, a left upward displacement (greater maximal effect, lesser effective concentration 50% and a steeper slope) compared with those obtained in the unsensitized group. These results indicate the existence of a non-specific increase in responsiveness in the airway smooth muscle from sensitized animals. PMID- 2569519 TI - Comparative effects of azapropazone on cellular events at inflamed sites. Influence on joint pathology in arthritic rats, leucocyte superoxide and eicosanoid production, platelet aggregation, synthesis of cartilage proteoglycans, synovial production and actions of interleukin-1 in cartilage resorption correlated with drug uptake into cartilage in-vitro. AB - Azapropazone (APZ) has been compared with standard NSAIDs in title systems to establish aspects of its mode of action on cellular events at inflamed sites. APZ (150 mg kg-1 day-1) given for 10-13 days exhibited a reduction in joint pathology in established adjuvant arthritis in rats comparable with that of indomethacin (2 mg kg-1 day-1) and clobuzarit (20 mg kg-1 day-1). APZ was shown to be a potent inhibitor of the production of leucocyte superoxide and synovial interleukin-1 (IL-1)-like activity and stimulated articular cartilage proteoglycan synthesis, but was ineffective as an inhibitor of platelet aggregation or IL-1 induced cartilage degradation in-vitro. These in-vitro effects may have relevance to the mode of action of this weak inhibitor of prostaglandin synthesis. PMID- 2569520 TI - Isolation and characterization of Pelamis platurus (yellow-bellied sea snake) postsynaptic isoneurotoxin. AB - Pelamis platurus (yellow-bellied sea snake) venom contains several neurotoxins, the major toxin, which is most toxic, and two other isotoxins. The second most toxic neurotoxin (Pelamis toxin b) was isolated and characterized. It contains 60 amino acid residues with only one residue difference from the major toxin, Pelamis toxin a. The difference is at the tenth amino acid residue from the acid terminal. The isoelectric point of toxin b is 8.7. Raman spectroscopic examination of toxin b indicates that the toxin contains a considerable amount of antiparallel beta-structure, beta-turn, and random coil without alpha-helix as the amide I band appears at 1673 cm-1 and the amide III band at 1246 cm-1. Circular dichroic studies also indicate a typical beta-sheet structure. The Pelamis toxin b is a typical postsynaptic neurotoxin as it binds to the acetylcholine receptor competitively with a well known toxin, alpha-bungarotoxin. The LD50 of toxin b is 0.185 microgram g-1 in mice by intravenous injection, indicating high toxicity of a postsynaptic neurotoxin. PMID- 2569522 TI - 3-Amino-1-hydroxypropylidene-1,1-diphosphonate (APD): a novel enhancer of rectal cefoxitin absorption in rats. AB - The promoting action of the calcium chelating compound EDTA on intestinal drug absorption is supposed to be based on Ca2+ depletion, inducing widening of tight junctions. The aim of the present study was to evaluate the effects of the calcium-binding agent 3-amino-1-hydroxypropylidene-1,1-diphosphonate disodium salt (APD) on rectal cefoxitin absorption in rats. The extent of rectal cefoxitin absorption was enhanced by 0.5 to 6% w/v of APD, on rectal infusion as well as on bolus delivery, the latter regimen tending to result in lower bioavailabilities. A maximal cefoxitin bioavailability of 85 +/- 10% was achieved by infusion with 4% w/v of APD, compared with 14 +/- 12% without APD. PMID- 2569521 TI - Effects of K+-canrenoate on the development of DOCA-salt hypertension. AB - The effects of K+-canrenoate, a digoxin antagonist, on the role of digoxin-like factor in the development of DOCA-salt hypertension has been examined. DOCA-salt rats treated with 66 mg kg-1 day-1 of K+-canrenoate (s.c.) presented a lower increase in blood pressure (P less than 0.01), less cardiac hypertrophy (P less than 0.05) and hypokalaemia (P less than 0.05) than non-treated DOCA-salt rats. K+-canrenoate treatment did not lead to significant changes in urinary volume, Na+ and K+ urinary excretion or suppression of plasma renin activity in DOCA-salt rats. None of the parameters were significantly different between uninephrectomized-salt rats treated or non-treated with K+-canrenoate. These data suggest a role for digoxin-like factor in DOCA-salt hypertension. However, the non-normalization of blood pressure observed in K+-canrenoate DOCA-salt treated rats indicates that other factors contribute to the initiating mechanisms in this type of hypertension. Moreover, these data suggest that digoxin-like factor plays no role in the suppression of plasma renin activity induced by DOCA and salt treatment. PMID- 2569523 TI - Continuous shear rheometry of o/w emulsions; control of evaporation in cone/plate geometry. AB - Volatile solvents may evaporate during cone/plate viscometry so that false rheograms develop. This surface evaporation was prevented in a cod-liver oil-in water emulsion stabilized with zanthoxylum gum by layering a film of cod-liver oil on the exposed surface of the emulsion test sample. The oil layer effectively prevented evaporation and did not alter significantly the rheological behaviour of the test material. PMID- 2569524 TI - Reduction by acetylsalicylic acid of paracetamol-induced hepatic glutathione depletion in rats treated with 4,4'-dichlorobiphenyl, phenobarbitone and pregnenolone-16-alpha-carbonitrile. AB - The role of enzyme induction in the reduction by acetylsalicylic acid (ASA) of paracetamol-induced hepatic glutathione (GSH) depletion has been studied in rats. Administration of an overdose of paracetamol to control rats resulted in an appreciable decrease of GSH concentration. Pretreatment with the enzyme inducers phenobarbitone, 3-methylcholanthrene (3-MC), pregnenolone-16-alpha-carbonitrile (PCN) and 4,4'-dichlorobiphenyl (4,4'-DCB) significantly potentiated the paracetamol-induced depletion of GSH. Simultaneous administration of an equimolar dose of ASA resulted in a reduction of the paracetamol-induced depletion of GSH in all instances except for those rats that were not pretreated and those given 3 MC. Benorylate, the ASA ester of paracetamol, depressed rat liver GSH to levels comparable to those produced by the combination of paracetamol and ASA. ASA itself caused only minor changes in liver GSH concentrations. The results demonstrate that ASA causes a diminution of paracetamol-induced GSH depletion in rats with phenobarbitone type of enzyme induction. Inhibition of the formation of the reactive metabolite of paracetamol or reduction of the absorption rate of paracetamol seem to be unlikely as mechanisms underlying the ASA-induced effect. An ASA-mediated effect via changes of the hepatic thiol status is proposed. PMID- 2569525 TI - L-ascorbic acid produces hypoglycaemia and hyperinsulinaemia in anaesthetized rats. AB - L-Ascorbic acid (Vitamin C) produced a marked reduction in the blood glucose concentration following intravenous injection (20-100 mg kg-1) to anaesthetized rats. This hypoglycaemic effect was accompanied by an increase in plasma insulin concentration. D-ascorbic acid produced a similar hypoglycaemic effect. PMID- 2569526 TI - RS-61756-007: a potent and selective thromboxane receptor (TP) agonist. AB - The activity of RS-61756-007 (methyl-9-oxo, 15 alpha-hydroxy, 16-phenoxy, 17,18,19,20-tetranor prosta 4,5, 13(E) trienoate, 4,5,6(R), 8(R)) has been assessed at prostanoid receptors (DP, EP1, EP2, FP, IP and TP) in-vitro. The activity profile of RS-61756-007 resembles that of U46619, in that agonism was observed at TP and to a lesser extent at FP receptors, but there was no activity at the remaining subtypes. The actions of both RS-61756-007 and U46619 were antagonized in a similar manner by the TP antagonist SQ 29,548. We conclude that RS-61756-007 is a highly potent TP agonist. PMID- 2569527 TI - The effects of dipyridamole and indomethacin on methotrexate cytotoxicity in LoVo human colon cancer cells. AB - Dipyridamole and indomethacin were studied for their effects in the in-vitro response of LoVo colon cancer cells to methotrexate (MTX) using a dye elution method. Dipyridamole 0.5-5 micrograms mL-1 or indomethacin 1 microgram mL-1 alone had little or no effect on cell growth. The tumour cells were refractory to even high concentrations of MTX (2.5-10 micrograms mL-1) alone or with indomethacin 1 microgram mL-1. In contrast, dipyridamole 0.5-5 micrograms mL-1 sensitized the cells to MTX 5 micrograms mL-1 (their growth was reduced by 25 to 69%), possibly by inhibiting thymidine salvage. PMID- 2569528 TI - Serum glucose and insulin levels in normotensive (WKY) and spontaneously hypertensive (SH) rats during and after the cessation of continuous (10 day) clonidine infusion. AB - The concentrations of glucose were elevated whereas those of insulin were decreased in the sera of normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive (SH) rats on day 10 of a continuous subcutaneous infusion of clonidine (10 micrograms kg-1 h-1 for 10 days). Fifteen to 18 h after cessation of infusion, the glucose levels of both strains had fallen to below those of the respective controls whereas the insulin levels remained suppressed. As such the hypoglycaemia may be related to the general increase in metabolic requirements associated with the post-infusion 'withdrawal syndrome'. PMID- 2569529 TI - 2-[125I]iodomelatonin binding sites in hamster and chick exhibit differential sensitivity to prazosin. AB - Binding of 2-[125I]iodomelatonin to putative receptors for melatonin has been well documented in chicken and hamster tissues. Differences in the potency of N acetyl-5-hydroxytryptamine in these tissues suggested distinct central and peripheral binding sites. Prazosin was found to be much more potent (Ki = 6.5 nM) in hamster hypothalamus than in chick retina (Ki = 740 nM). N-acetyl-5-HT also was more potent in hamster hypothalamus (Ki = 14 nM) than in chick retina (Ki = 1050 nM). A comparison of hamster hypothalamus to chick forebrain revealed a similar difference in potency for prazosin. In light of these findings, the dissimilarity between the potencies of N-acetyl-5-HT and prazosin in hamster and chick would appear to be due to a species difference as the central and peripheral sites seem alike within either species. PMID- 2569530 TI - A study of beta-adrenoceptors in rat lung parenchymal strip. AB - The aim of the present study was to characterize the beta-adrenoceptor population in rat lung strip. For this purpose, Schild plots were obtained for the beta adrenoceptor antagonists atenolol (beta 1-selective), butoxamine (beta 2 selective) and propranolol (non-selective), using three different agonists:isoprenaline (non-selective), salbutamol (beta 2-selective) and noradrenaline (beta 1-selective). The slopes of these Schild plots were close to the theoretical value of unity, and pA2 values determined with isoprenaline, salbutamol and noradrenaline as agonists were: for propranolol, 7.86 +/- 0.22, 7.72 +/- 0.15 and 7.89 +/- 0.23; for atenolol, 5.19 +/- 0.05, 5.33 +/- 0.07 and 5.47 +/- 0.22 and for butoxamine, 6.31 +/- 0.11, 6.34 +/- 0.03 and 5.99 +/- 0.23, respectively. These data suggest that pharmacological responses of rat isolated lung strip to beta-adrenoceptor agents are mediated by a homogeneous population of beta 2-adrenoceptors, although the presence of a minor population of beta 1 adrenoceptors undetected by the agonists used cannot be excluded. PMID- 2569531 TI - Cefazaflur: kinetics of hydrolysis in aqueous solution, acid dissociation constant and alkaline decomposition to fluorescent products. AB - The values of the pseudo-first order hydrolysis rate constants in the pH range 1 to 10, and the pKa, were determined for cefazaflur in aqueous solution at 37 degrees C and ionic strength 0.2 M. A fluorimetric assay, based on alkaline hydrolysis at 100 degrees C, was also developed for this compound. The results are consistent with previously reported related properties of other monoprotic cephalosporins. PMID- 2569532 TI - Studies on the cardiovascular actions of central histamine H1 and H2 receptors. AB - Histamine administered i.c.v. to conscious freely moving rats results in dose related pressor responses and bradycardia. The selective H1 agonist pyridylethylamine (PEA) and the H2 agonist impromidine (IMP) were utilized to characterize the central receptor subtypes involved in the central cardiovascular actions of histamine. Blood pressure and heart rate were monitored directly via indwelling carotid catheters, and drugs were administered i.c.v. through permanently implanted cannulas. Central administration of PEA or IMP produced dose-dependent pressor responses and bradycardia. The H1 antagonist chlorpheniramine blocked the pressor response, but not the bradycardia produced by PEA. In contrast, the H2 antagonist BMY-25405 blocked both the increase in blood pressure and the bradycardia induced by IMP. Crossover experiments were carried out to examine the specificity of the antagonists' actions. Chlorpheniramine failed to block the actions of IMP and BMY-25405 likewise was ineffective in blocking the actions of PEA. These results suggest that both central H1 and H2 receptors mediate the central pressor effects of histamine, and that PEA and IMP are selectively acting at their respective receptors in the brain. The heart rate effects appear to be mediated directly by central H2 actions, as BMY-25405 was capable of blocking the heart rate changes. The finding that IMP is less potent in the central nervous system, compared to its known greater potency in the periphery, suggests that peripheral and central H2 receptors may be pharmacologically distinct. PMID- 2569533 TI - Differential effects of somatostatin on atrial and ventricular contractile responses in guinea pig heart: influence of pretreatment with islet-activating protein. AB - The effects of somatostatin on the contractile response of guinea pig cardiac preparations were investigated and compared with those of carbachol and adenosine. Somatostatin produced a concentration-dependent negative inotropic effect in the left atria, which was accompanied by a decrease in action potential duration. The maximum decrease in contractility which was obtained at 3 x 10(-6) M was around 40% of the predrug control values and far less than those produced by carbachol and adenosine. Somatostatin failed to produce inotropic effect on the papillary muscle and did not influence the spontaneously beating rate of the right atria. In the papillary muscles, however, somatostatin inhibited the positive inotropic effect of isoproterenol in a concentration-dependent manner as did carbachol and adenosine. In addition, somatostatin caused a significant inhibition of the isoproterenol-induced increase in cyclic AMP levels without affecting the basal level of cyclic AMP. In the papillary muscle, the inhibitory effect of somatostatin on the positive inotropic response to isoproterenol was significantly attenuated by pretreatment with islet-activating protein, and was significantly antagonized by the somatostatin antagonist cyclo[7-aminoheptanoyl Phe-D-Trp-Lys-Thr(Bzl)]. These results suggest that somatostatin receptors in guinea pig ventricular muscles are coupled with adenylate cyclase via islet activating protein-sensitive GTP-binding protein, whereas the negative inotropic effect of somatostatin in the left atria might be mediated by a subtype of somatostatin receptors which is different from that in the ventricle. PMID- 2569534 TI - Delayed rescue of N-methyl-D-aspartate receptor-mediated neuronal injury in cortical culture. AB - The present study explored the neuroprotective efficacy of delayed manipulations performed after completion of an excitotoxic insult. Many cultured murine cortical neurons that would otherwise die after exposure to 500 microM N-methyl-D aspartate (NMDA) or glutamate could be rescued by the late addition of NMDA antagonists to the bathing medium. D-2-Amino-5-phosphonovalerate (D-APV), MK-801 (5-methyl-10, 11-dihydro-5H-dibenzo[a,d]cyclohepten-5, 10-imine maleate) and dextrorphan all produced concentration-dependent neuroprotection, with EC50 about 10, 0.3 and 3 microM, respectively, and similar efficacy. The fraction of the doomed neuronal population that could be rescued depended on the time interval between washout of NMDA and antagonist addition, starting at a maximum of 40 to 80% with immediate addition, and decaying to zero after 30 min. D-APV only needed to be present for 30 min for near maximal protection. The ability of NMDA antagonists to rescue doomed neurons was not mimicked by several other drugs: 1 mM gamma-D-glutamyl-aminomethyl sulfonate, 1 mM L-glutamate diethyl ester, 10 microM 6-nitro-7-cyano-quinoxaline-2,3-dione, 1 mM secobarbital, 100 microM diphenylhydantoin, 10 microM nifedipine or 3 microM tetrodotoxin. The broad spectrum glutamate antagonist kynurenate had a protective action similar to that of D-APV, but when added to D-APV did not improve neuroprotection. Neurons could also be rescued by the delayed removal of extracellular calcium for 30 min after exposure to NMDA. In contrast, replacement of sodium with choline actually enhanced resultant neuronal damage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569535 TI - Atrial natriuretic peptide-induced stimulation of cyclic GMP formation by isolated bovine luteal cells. AB - Incubation of bovine luteal cells with an atrial natriuretic peptide (rat atriopeptin II, rAP-II) did not affect hCG-stimulated or basal cyclic AMP accumulation and progesterone production, but cyclic GMP formation was stimulated by rAP-II in a dose-dependent manner, being maximally stimulatory in the nanomolar range. This stimulatory influence of rAP-II on cyclic GMP formation results from a specific stimulation of particulate guanylate cyclase. We suggest that, although rAP-II mediated cyclic GMP formation can be demonstrated in bovine luteal cells, there appear to be no acute effects of the atrial peptide on the regulation of progesterone production by these cells. PMID- 2569537 TI - Genetic diversity of simian immunodeficiency virus. AB - We have demonstrated that the genetic diversity of simian immunodeficiency virus from African green monkeys (SIVagm) is much greater than that observed previously for individual HIV-1, HIV-2, or SIVmac isolates. Extensive genetic variation among SIVagm isolates and the high prevalence of green monkey infection without disease suggest that the virus has been in the green monkey population for a long time. We have also demonstrated that SIV from a sooty mangabey monkey (isolate SMM-7) is closer to SIVmac and HIV-2 than to HIV-1 and SIVagm. The extensive genetic diversity of SIVagm and the relatedness of SIVsmm to HIV-2 warrant continued examination of SIVagm and SIVsmm isolates from dispersed geographic regions. SIV strains much more closely related to HIV-1, HIV-2, or SIVmac may be found which would be reasonable candidates for recent cross-species transmission. PMID- 2569536 TI - Leucocyte membrane antigens on mouse granulated metrial gland cells. AB - An indirect immunofluorescence technique has been used to study the expression of leucocyte membrane antigens on mouse granulated metrial gland (GMG) cells. GMG cells isolated from cultured metrial gland explants and GMG cells in cryostat sections of uterine implantation sites were examined. GMG cells were found to express both the asialo-GM1 antigen and the Thy-1 antigen. GMG cells did not express the Lyt-1, Lyt-2, Mac-1, L3T4 or IgM antigens. These results provide new evidence that GMG cells are a type of NK cell. PMID- 2569538 TI - Metabolism and hepatorenal toxicity due to repeated exposure to styrene in spontaneously hypertensive rats (SHR). AB - Groups of adult male rats (5 rats per group), either normotensive (WKY) or spontaneously hypertensive (SHR), were exposed by inhalation to 0, 821, and 3018 ppm styrene, 5 h per day for 3 consecutive days. After the exposure, the urines were collected for 24 h and the animals were then sacrificed. The various biochemical parameters of hepatorenal toxicity due to styrene as well as its urinary metabolites were measured. Hepatotoxicity due to styrene was not further increased at any exposure level due to hypertension. However, repeated exposure of SHR rats to 3018 ppm styrene showed significant increases in the urinary excretion of gamma-glutamyl transpeptidase, proteins, and volume of urine, compared to WKY treated rats, whereas no such changes were observed due to repeated exposure to 821 ppm styrene. Studies of in vivo metabolism of styrene at higher exposure level showed significant decrease in the urinary excretion of mandelic, phenylglyoxylic, and hippuric acids in SHR rats compared to WKY-treated rats, suggesting an inhibition of deactivation of styrene reactive intermediate involving the epoxide hydrase pathway due to hypertension. At the same time, a significant increase in the urinary excretion of a potential nephrotoxic metabolite of styrene (e.g., mercapturates or thioethers) was observed in SHR treated rats when compared to WKY-treated rats. These results demonstrate that spontaneous hypertension has the potential to further increase the nephrotoxicity due to repeated exposure to styrene, and the metabolism of styrene plays an important role in modifying such toxicity in the hypertensive state. PMID- 2569539 TI - Effect of nerve growth factor on the synthesis of amino acids in PC12 cells. AB - Radioactive short-chain fatty acids preferentially label glutamine relative to glutamate in brain due to compartmentation of glutamine and glutamate. To determine whether this phenomenon occurs in a single cell culture model, we examined the effect of fatty acid chain length on the synthesis as well as pool size of selected amino acids in rat pheochromocytoma PC12 cells, a cell culture model of the large glutamate compartment in neurons. Intracellular 14C-amino acids were quantitated by HPLC, and the incorporation of [U-14C]-glucose, [1-14C] butyrate, [1-14C]-octanoate, and [1-14C]-palmitate into five amino acids was measured in native and NGF-treated PC12 cells. NGF pretreatment decreased the intracellular concentration of amino acids as did addition of fatty acids but these effects were not additive. Specific activities of amino acids in native cells labelled by 14C-octanoate were 1,300 DPM/nmol, 490 DPM/nmol, 200 DPM/nmol, and 110 DPM/nmol for glutamate, aspartate, glutamine, and serine, respectively. No radioactivity was detected in alanine. Similar specific activities were noted when 14C-butyrate was the precursor; however, there was at least 5-fold less if 14C-palmitate was the precursor. Pretreatment of cells with NGF decreased the specific activity of amino acids by 25-65%. Specific activities of amino acids synthesized from 14C-glucose decreased in the following order: glutamate, 1,640 DPM/nmol; aspartate, 1,210 DPM/nmol; alanine, 580 DPM/nmol; glutamine, 275 DPM/nmol; and serine, 80 DPM/nmol for native cells. NGF pretreatment decreased the specific activities of glutamate and glutamine, but not of the other 3 amino acids. The preferred precursor for glutamate synthesis in native PC12 cells was glucose followed by octanoate, butyrate and palmitate (16:6:3:1). PMID- 2569540 TI - Neurotransmitter phenotype plasticity in cultured dissociated adult rat dorsal root ganglia: an immunocytochemical study. AB - Culturing sympathetic ganglion neurons in vitro may modify phenotypic expression of some neurotransmitters. For dorsal root ganglia (DRG), contradictory results have been reported; most studies have used immature material. We have therefore performed a detailed immunocytochemical analysis of the transmitter content of cultured adult rat DRG neurons. To demonstrate possible modifications of neurotransmitter phenotypes, we have compared the results obtained with the same techniques on neurons cultured for 3 days and on freshly dissociated DRG cells. Also, the transmitter profile of cultured neurons was compared with that known from in situ studies. Out of 22 antigens studied, 20 were detected in cultured DRG neurons. All of them were expressed in small and/or intermediate-sized cells. Large neurons only contained CGRP, VIP, NPY, beta-END, ENK, and GABA. The percentage of immunostained neurons varied for the various antisera: less than 10% of cultured neurons were positive for ENK, beta-LPH, beta-END, DYN, VASO, and OXY; 10-30% for SOM, CCK, CAT, and SP; and greater than 30% for NPY, CRF, GLU, NT, VIP, GABA, GRP, CGRP, 5-HT, and TRH. In the latter two groups of transmitters (except CGRP), the proportion of immunoreactive neurons was by far larger in cultured than in freshly dissociated DRG. The most pronounced (greater than 25%) increase in the proportion of positively stained neurons after culturing was observed for the GRP, CRF, TRH, and 5-HT antisera. Serotonin was the only transmitter identified in cultured but not in freshly dissociated cells. These data indicate, on one hand, that various antigens, for example, CAT, GABA, NT, TRH, NPY, beta-LPH, and beta-END, which up to now have not been described in DRG in situ, can be detected immunocytochemically a few hours after dissociation of adult rat DRG. On the other hand, several transmitters, for example, VIP, NPY, SP, GABA, GLU, NT, GRP, CRF, TRH, and 5-HT, are expressed in a significantly higher proportion of cells in cultured than in freshly dissociated preparations. This might reflect a change in the phenotypic expression of transmitters due to the new environment generated by the culture conditions, a hypothesis that can be tested by measuring specific mRNA levels. Moreover, considering the plasticity and multipotentiality of their transmitter phenotype, cultured adult DRG neurons might represent an interesting material for autografts into the injured central nervous system. PMID- 2569541 TI - Studies on lymphocyte subpopulations and NK cell activities in epileptic patients. AB - The subpopulations of T lymphocytes, B lymphocytes and macrophages of the peripheral blood in 58 cases of epilepsia were studied with McAb by IFA method. The activity of natural killer (NK) cells of peripheric blood monocyte was investigated by 51Cr-releasing method. 22 healthy donors of about the same age served as controls. The results showed that the percentage of T3 and T4 lymphocytes in epileptic patients was decreased, and the ratio of T4/T8 was markedly reduced as compared to control group. The percentage of T8 lymphocytes was increased. There were no significant changes in B lymphocytes and macrophages. The NK cell activity also showed decrease. The results suggested abnormality in cellular immunity in epilepsia, which may be involved in the pathogenetic mechanism of the disease. PMID- 2569542 TI - Continuing medical education. PMID- 2569543 TI - Hemodynamic benefits of long-term bunazosin (alpha-1 blocker) therapy in rats with myocardial infarction and heart failure. AB - To determine whether bunazosin (alpha 1-adrenoceptor blocking agent) can alter the hemodynamic profile of chronic heart failure secondary to myocardial infarction, the drug was administered to Wistar rats 4 weeks after coronary ligation, and continued for 4 weeks. In rats without bunazosin treatment, the left ventricular end-diastolic pressure (LVEDP) and the total vascular resistance index (TVRI) increased as a function of infarct size, while the cardiac index (CI) decreased. But in infarcted rats with bunazosin treatment, the mean aortic pressure and TVRI were reduced, the LVEDP was modestly lessened, and the CI was maintained. The greatest increase in CI after the treatment occurred in rats with infarcts of small and moderate size. Thus, long-term therapy with bunazosin improved LV dysfunction, relative to the size of infarction. This study suggests the beneficial effects of bunazosin therapy in patients with chronic heart failure. PMID- 2569544 TI - Effects of mafoprazine, a phenylpiperazine derivative, on the central dopaminergic system. AB - The effects of mafoprazine, a new phenylpiperazine derivative, on the central dopaminergic system were studied. Mafoprazine, like chlorpromazine and haloperidol, reduced the apomorphine-induced cage-climbing behavior in mice, emesis in dogs and stereotyped behavior in monkeys; methamphetamine-induced hyperlocomotion and group toxicity in mice; and agitation in rats. Mafoprazine inhibited the unilateral circling behavior induced by methamphetamine and apomorphine in rats with 6-hydroxydopamine-induced lesions in the unilateral nigrostriatal neuronal tract. The potency of mafoprazine in these experiments was almost equal to that of chlorpromazine and about one-tenth that of haloperidol. The cataleptogenic activity of mafoprazine was lower than those of chlorpromazine and haloperidol. Mafoprazine potentiated clonidine-induced hypothermia. These results suggest that mafoprazine has a relatively selective postsynaptic dopamine D2-receptor blocking action in the nucleus accumbens compared with chlorpromazine and haloperidol and suggest that mafoprazine also has alpha 2-adrenoceptor stimulating actions. PMID- 2569545 TI - Selective blockade of dopamine D-1 receptor by SCH 23390 affects dopamine agonist binding to 3H-spiperone labeled D-2 receptors in rat striatum. AB - This study investigated the effects of selective blockade of dopamine D-1 receptors by SCH 23390 and selective stimulation of the receptors by SKF 38393 on the binding characteristics of 3H-spiperone labeled D-2 receptors in rat striatum. Selective blockade of D-1 receptors by 50 nM SCH 23390 significantly decreased the affinity of dopamine agonist for 3H-spiperone labeled D-2 receptors, but did not influence dopamine antagonist binding to D-2 receptors. Selective stimulation of D-1 receptors by SKF 38393 (100 nM) did not affect either dopamine agonist or antagonist binding to D-2 receptors. The characteristics of the effect of SCH 23390 on dopamine agonist binding to D-2 receptors was similar to those of GTP, but different from those of sodium ion. This effect could not be due to a direct modification of D-2 receptors by SCH 23390. Pertussis toxin (IAP) treatment significantly decreased the affinity of dopamine agonist for D-2 receptors and reduced the abilities of both SCH 23390 and GTP to decrease the affinity of dopamine agonist for D-2 receptors. These results suggest, therefore, putative interregulatory mechanism between dopamine D 1 and D-2 receptors and the possible involvement of a pertussis toxin sensitive protein in this mechanism. PMID- 2569546 TI - [Sensitivity of opportunistic-pathogenic microflora isolated from humans to antibiotics before and after a stay in a hermetically closed chamber]. AB - Antibiotic sensitivity of opportunistic microflora of men before and after exposure to an enclosed environment for as long as 2-8 to 96-175 days was investigated. It was found that after exposure the spectrum of opportunistic enterobacteria resistant to antibacterial drugs was enlarged. The results of serotyping of the isolated bacteria showed that one of the pathways that led to an increase in the number of antibiotic-resistant microorganisms was individual variations in the intestinal automicroflora which were characterized by a decrease in the amount of some species of enterobacteria and an increase in the amount of other species. PMID- 2569548 TI - Sudden collapse of a young, healthy man. PMID- 2569547 TI - [Combined effects of increased levels of carbon dioxide and environmental temperature on the thermal status of humans in a hermetically closed compartment]. PMID- 2569549 TI - Acute lymphoblastic leukemia: current controversies, future directions. PMID- 2569550 TI - Routine formal fetal movement counting and risk of antepartum late death in normally formed singletons. AB - The routine recommendation to women to count fetal movements daily during late pregnancy for the prevention of antepartum late fetal death in normally formed singletons has been evaluated. 68,000 women were randomly allocated within thirty three pairs of clusters either to a policy of routine counting or to standard care, which might involve selective use of formal counting or informal noting of movements. Antepartum death rates for normally formed singletons were similar in the two groups, regardless of cause of prior risk status. Despite the counting policy, most of these fetuses were dead by the time the mothers received medical attention. The study does not rule out a beneficial effect, but at best, the policy would have to be used by about 1250 women to prevent 1 unexplained antepartum late fetal death, and an adverse effect is just as likely. In addition, formal routine counting would use considerable extra resources. PMID- 2569551 TI - Prediction of severity of Gaucher's disease by identification of mutations at DNA level. AB - The polymerase chain reaction was used to detect four mutations in the DNA of 47 unrelated patients with type I Gaucher's disease (94 Gaucher's disease alleles). Two of the mutations, 1226 and 1448, and a new mutation (XOVR) representing cross over between the glucocerebrosidase gene and its closely linked pseudogene, were found. There were five genotypes--namely, 1226/1226, 1226/1448, 1226/XOVR, 1226/?, and ?/? (where "?" indicates that none of the four known mutations was present). Severity of the disease was assessed with a scoring index according to age at diagnosis and extent of organ involvement. Mutation 1226 was associated with a mild clinical phenotype, and mutation 1448 with a more severe phenotype. Mutation 1226 is the most common cause of Gaucher's disease in Jewish patients. PMID- 2569552 TI - Multiple genetic alterations in distal and proximal colorectal cancer. AB - Multiple genetic alterations were investigated in colorectal cancer, including changes in DNA content, mutations in ras oncogenes, and deletions involving chromosomes 5, 17, and 18. A non-random association of deletions and mitotic abnormalities by site was seen, with both types of alterations occurring significantly more frequently in distal tumours. In contrast, the frequency of c Ki-ras mutations did not differ between proximal and distal cancers. In addition, deletions were significantly associated with each other and with change in DNA content. The data provide strong support for the hypothesis that proximal and distal colon carcinoma might differ in the genetic mechanisms in their initiation and/or progression. PMID- 2569553 TI - S-methylation in motorneuron disease and Parkinson's disease. AB - Thiolmethyltransferase activity has been measured in newly diagnosed, untreated patients with idiopathic Parkinson's disease and motorneuron disease, and in normal volunteers. In Parkinsonian patients, mean thiolmethyltransferase activity was low (300 U/mg protein [SD 96]) compared with that in controls (947 [409]), whereas activity was high in patients with motorneuron disease (2077 [825]). PMID- 2569554 TI - Anti-factor-VIII inhibitors in haemophilia. PMID- 2569555 TI - Brain damage and open-heart surgery. PMID- 2569556 TI - Lessons in the surgical treatment of epilepsy. PMID- 2569557 TI - Informing patients about clinical disagreement. PMID- 2569558 TI - Hormone replacement therapy and cancer: is there cause for concern? PMID- 2569559 TI - Aggressive management of severe closed head trauma: time for reappraisal. AB - The records of 187 patients (age 6 months to 82 years) who sustained severe closed head injuries and were admitted to a neurosurgical unit between 1983 and 1987 were retrospectively reviewed. The aim was to develop a scale for selecting patients whose death could be predicted with certainty at an early stage and who should therefore not receive prolonged intensive care. The scale was prospectively tested on 52 comparable patients admitted during 1988 and was found to be accurate with no falsely pessimistic predictions. The score is based mainly on clinical observations within 12 h of presentation. PMID- 2569560 TI - Outcome in newborn babies given anti-varicella-zoster immunoglobulin after perinatal maternal infection with varicella-zoster virus. AB - 281 newborn babies whose mothers had chickenpox and 25 whose mothers had herpes zoster during the perinatal period were investigated. IgG antibody was present at birth in all babies born more than 7 days after the onset of maternal chickenpox. When the mother's rash appeared 7-3 days before delivery progressively fewer babies were born with antibody, and no infant born less than 3 days after the onset had antibody at birth. Infants were given 100 or 250 mg of anti-varicella zoster immunoglobulin (VZIG) shortly after birth or the onset of post-natal maternal chickenpox. 1 infant died, without neonatal varicella. Of the 280 surviving infants 169 (60%) were infected--134 (48%) with chickenpox and 35 (13%) without clinical features. The clinical attack rate was highest (60%) in infants whose mothers had chickenpox between 7 days before and 7 days after delivery. Chickenpox was severe in 19 infants, 16 of whom were exposed to maternal chickenpox between 4 days before and 2 days after delivery. There was no evidence that a severe outcome was associated with transplacentally acquired infection. Perinatal maternal herpes zoster did not cause neonatal infection. VZIG should be given to infants at risk, including those whose mothers have chickenpox during the last 7 days of pregnancy. PMID- 2569561 TI - Medication errors in neonatal and paediatric intensive-care units. AB - In a 4-year prospective quality assurance study, 315 iatrogenic medication errors were reported among the 2147 neonatal and paediatric intensive-care admissions, an error rate of 1 per 6.8 admissions (14.7%). The frequency of iatrogenic injury of any sort due to a medication error was 66/2147 (3.1%)--1 injury for each 33 intensive-care admissions. 33 (10.5%) errors were potentially serious, 32 (10.2%) caused mild patient injuries, and 1 patient had acute aminophylline poisoning after receiving five intravenous doses of the drug at a dosage ten times higher than prescribed, owing to a calculation error during dilution. A longitudinal monitoring system helps to identify iatrogenic complications due to medication errors and may help in implementing preventive measures. PMID- 2569563 TI - RAST screening for antibodies to anaesthetics. PMID- 2569562 TI - Setting the price of essential drugs: necessity and affordability. AB - A method has been developed for fine tuning the selection of drugs to improve cost recovery, to promote appropriate drug use, and to make more drugs more affordable. This scheme is based on a classification of drug necessity (vital, essential, non-essential) and on the relative cost of complete courses of treatment so that expensive drugs can be subsidised by marking up inexpensive ones. PMID- 2569564 TI - Cocaine and morphine withdrawal. PMID- 2569565 TI - Susceptibility of human herpesvirus 6 to acyclovir and ganciclovir. PMID- 2569566 TI - Photodynamic therapy. PMID- 2569567 TI - Single treatment of invasive fascioliasis with triclabendazole. PMID- 2569568 TI - Fine-needle biopsy in mammographically detected non-palpable lesions. PMID- 2569569 TI - Genetics of coronary heart disease. PMID- 2569570 TI - Evaluation of screening tests for urinary infection in hospital patients. PMID- 2569571 TI - Continuous infusion of ceftazidime in cystic fibrosis. PMID- 2569572 TI - High prevalence of thyroid nodule in area of radioactive fallout. PMID- 2569573 TI - AIDS and academic boycotts. PMID- 2569574 TI - HIV testing in 25% of Swedish population aged 16-44. PMID- 2569575 TI - Percutaneous sclerosis of gallbladder. PMID- 2569576 TI - Air bubbles in insulin pens. PMID- 2569577 TI - Treatment of malignant melanoma by single thermal neutron capture therapy with melanoma-seeking 10B-compound. PMID- 2569578 TI - Campylobacter pylori, gastrin, and duodenal ulcer. PMID- 2569579 TI - Omeprazole, campylobacter pylori, and duodenal ulcer. PMID- 2569580 TI - Acute neuroborreliosis in a patient previously infected with Borrelia burgdorferi. PMID- 2569581 TI - Use of recombinant tissue plasminogen activator before balloon angioplasty. PMID- 2569583 TI - Cervical smears: a questionable practice? PMID- 2569582 TI - Tolerance of two brands of praziquantel. PMID- 2569585 TI - Non-surgical treatment for pyloric stenosis. PMID- 2569584 TI - Effective cytological sampling. PMID- 2569586 TI - Mumps meningitis following measles, mumps, and rubella immunisation. PMID- 2569587 TI - Complement activation and high-dose of interleukin-2. PMID- 2569588 TI - Something new in liver cirrhosis epidemiology. PMID- 2569589 TI - Simultaneous outbreaks of contrasting drug resistant classic and El Tor Vibrio cholerae O1 in Bangladesh. PMID- 2569590 TI - Dysgonic fermenter-2 meningitis simulating viral meningitis. PMID- 2569591 TI - Rising age limit for kidney donors? PMID- 2569593 TI - Heat illness and servicemen. PMID- 2569592 TI - Halofantrine and pruritus. PMID- 2569594 TI - Spinal toxocaral abscess. PMID- 2569595 TI - Ultimate in compulsive water drinking? PMID- 2569596 TI - Avoidable heatstroke in the armed forces. PMID- 2569597 TI - Female to male transmission of human immunodeficiency virus type 1: risk factors for seroconversion in men. AB - To determine the frequency and risk factors for female to male sexual transmission of human immunodeficiency virus type 1 (HIV-1), a prospective study was carried out in 422 men who had acquired a sexually transmitted disease (STD) from a group of prostitutes with a prevalence of HIV-1 infection of 85%. The initial seroprevalence of HIV among the men was 12%. 24 of 293 (8.2%) initially seronegative men seroconverted to HIV-1. Newly acquired infection was independently associated with frequent prostitute contact (risk ratio 3.2, 95% confidence interval 1.2-8.1), with the acquisition of genital ulcer disease (risk ratio 4.7, 95% confidence interval 1.3-17.0), and with being uncircumcised (risk ratio 8.2, 95% confidence interval 3.0-23.0). 96% of documented seroconversions occurred in men with one or both of the latter two risk factors. In a subgroup of 73 seronegative men who reported a single prostitute sexual contact, the frequency of HIV-1 infection was 8.2% during 12 weeks of observation. No man without a genital ulcer seroconverted. A cumulative 43% of uncircumcised men who acquired an ulcer seroconverted to HIV-1 after a single sexual exposure. These data indicate an extremely high rate of female to male transmission of HIV-1 in the presence of STD and confirm a causal relation between lack of male circumcision, genital ulcer disease, and susceptibility to HIV-1 infection. PMID- 2569599 TI - Neutrophil-mediated endothelial injury in haemolytic uraemic syndrome. AB - Neutrophil leucocytosis is associated with a poor outcome in the haemolytic uraemic syndrome (HUS). This study tested the hypothesis that neutrophils from HUS patients are activated and through release of their intracellular contents damage endothelium. The proportion of neutrophils adhering to endothelium in culture was twice as high for HUS patients' neutrophils as for control neutrophils (n = 12). In addition, these neutrophils induced endothelial injury, assessed morphologically by degradation of endothelial cell fibronectin. In an attempt to inhibit neutrophil adhesion and subsequent endothelial damage the hyperadhesive neutrophils from HUS patients were incubated with a CD18 antibody directed against the common beta chain of the leucocyte integrin molecules. The CD18 antibody was able to abrogate endothelial damage in four of the ten subjects studied. These observations suggest that the neutrophil is of prime pathophysiological importance in HUS, and that methods aimed at reducing neutrophil adhesion and neutrophil-mediated endothelial damage are likely to be beneficial. PMID- 2569598 TI - Rapid emergence of AIDS in Abidjan, Ivory Coast. AB - Between July and November, 1988, 1501 consecutive adult medical admissions to the two largest hospitals in Abidjan, Ivory Coast, were studied. The overall prevalence of infection with human immunodeficiency virus (HIV) types 1 and 2 was 43% in hospital A and 28% in hospital B. AIDS accounted for 19% and 9%, respectively, of medical admissions to the two hospitals, and for 33% of medical deaths in hospital A. The risk of death was significantly higher in HIV seropositive patients with acquired immunodeficiency syndrome (AIDS) (risk ratio 2.5, 95% confidence interval [CI] 2.0-3.2) and in HIV-positive patients who did not meet the AIDS case-definition (risk ratio 1.7, 95% CI 1.3-2.3) than HIV negative patients. The male/female ratio of the 265 cases of AIDS identified was 4.8. 50% of patients with AIDS were seropositive for HIV-1 only, 4% for HIV-2 only, and 46% reacted serologically to both viruses. The minimum incidence of AIDS in adult male and female residents of Abidjan (per million per year) is estimated at 1447 and 340 cases, respectively. AIDS in this West African city has emerged as a major public health problem. PMID- 2569600 TI - Trial of normobaric and hyperbaric oxygen for acute carbon monoxide intoxication. AB - The value of hyperbaric oxygen in the treatment of acute carbon monoxide intoxication was assessed in 629 adults who had been poisoned at home in the 12 h before admission to hospital. In patients without initial impairment of consciousness (group A) the effect of 6 h of normobaric oxygen (NBO) (group A0, n = 170) was compared with that of 2 h of hyperbaric oxygen (HBO) at 2 atmospheres absolute (ATA) plus 4 h NBO (group A1, n = 173). At the 1 month follow-up 66% of A0 and 68% of A1 patients had recovered. In patients with initial impairment of consciousness the effect of one session of HBO (group B1, n = 145) was compared with that of two sessions (group B2, n = 141); all group B patients also received 4 h of NBO. At 1 month of follow-up 54% group B1 and 52% group B2 patients had recovered. The 7 patients left with neuropsychiatric sequelae (3 B1, 4 B2) and the 4 who died (2 B1, 2 B2) had all presented with coma. HBO was not useful in patients who did not lose consciousness during carbon monoxide intoxication, irrespective of their carboxyhaemoglobin level, nor were two sessions of HBO in patients who sustained only a brief loss of consciousness. The prognosis is poorest for those presenting with coma; the trial needs to be pursued in this group of patients until the power of the study is sufficient to demonstrate the value or otherwise of HBO. PMID- 2569601 TI - Prenatal diagnosis of a red-cell enzymopathy: triose phosphate isomerase deficiency. AB - A child with triose phosphate isomerase deficiency was born to nonconsanguineous parents, and died at 13 months of age. The parents were both found to be heterozygous for this enzyme deficiency. At a subsequent pregnancy, analysis of fetal red blood cells obtained by cordocentesis at 19 weeks' gestation enabled prenatal diagnosis of the heterozygous state. This technique may allow diagnosis of other red-cell enzymopathies during the second trimester. PMID- 2569602 TI - Pathology of coronary angioplasty. PMID- 2569603 TI - Agrobureaucracy vs health. PMID- 2569604 TI - Sampling grossly benign breast biopsy specimens. PMID- 2569605 TI - Superstring theory. PMID- 2569606 TI - Fish oil revisited. PMID- 2569607 TI - Managing the sick doctor. PMID- 2569608 TI - Food-based oral rehydration salt solution for acute childhood diarrhoea. AB - The efficacy in acute childhood diarrhoea of oral rehydration therapy (ORT) based on staple foods (maize, millet, wheat, sorghum, rice, or potato) was compared with that of standard ORT based on glucose. 266 children aged 1-5 years, with a history of acute diarrhoea for 48 h or less, moderate to severe dehydration, and no complications, were assigned to treatment with one of the food-based oral rehydration salt solutions (ORS) or standard ORS. The mean stool output over the first 24 h of treatment in the group receiving standard ORS was significantly higher than that of any other treatment group, and the groups receiving food based ORT showed substantial reductions in stool output compared with the standard ORT group. Abnormalities in electrolyte concentrations were corrected in all treatment groups with similar efficiency. The digestibility of the food-based ORS was assessed by the stool pH, glucose content before and after acid hydrolysis, and osmolality; there were no significant differences between the standard ORS and food-based ORS groups. Food-based ORT should be more acceptable to users in developing countries since the mixtures are similar to traditional weaning foods and since, unlike standard ORT, it reduces stool output substantially. PMID- 2569609 TI - Endoscopic sphincterotomy in 1000 consecutive patients. AB - Between 1983 and 1988, endoscopic sphincterotomy was attempted on 1000 consecutive patients with a clinical diagnosis of bileduct stones in a centre with a policy to establish immediate bileduct drainage for retained stones. Endoscopic cholangiography was successful in 985 patients, of whom 782 had visible stones and 203 had a dilated bileduct but no visible stones. Endoscopic sphincterotomy was successful in 975 of these patients, with eventual bileduct clearance in 674 of 772 patients (87.3%) with visible stones; immediate bileduct drainage was achieved in 160 of the 161 patients (99%) in whom bileduct clearance failed at the first attempt. Overall, 771 of 797 patients (96.7%) with visible bileduct stones had successful bileduct clearance or drainage. Complications occurred in 6.9%, with a 30-day mortality rate of 1.2%, but procedure-related mortality was only 0.6%. PMID- 2569610 TI - Venous thrombosis and rifampicin. AB - In a retrospective analysis of clinically diagnosed and lower limb deep vein thrombosis (DVT) proven by contrast venography, DVT complicated admissions in 46 (3.4%) of 1366 adult patients treated in a tuberculosis hospital during 1986. Analysis of 7542 admissions during 1978-86 showed a relative risk of 4.74 in patients treated with regimens including rifampicin compared with other regimens. DVT was significantly more common in winter months and usually occurred within 2 weeks of treatment being started. This probable association between rifampicin and DVT does not contraindicate use of this drug, but measures to prevent DVT should be taken in inpatients receiving rifampicin. PMID- 2569611 TI - Salmonella enteritidis phage type 4 infection of broiler chickens: a hazard to public health. AB - The pericardial fluids and contents of caeca and spleens from 81 broiler chickens that had been condemmed at processing factories because of macroscopic pericarditis were examined for Salmonella species. 47 (58%) of these chickens yielded S enteritidis phage type (PT) 4. Viable counts of the organism in fluids from 6 of the most severely affected hearts ranged from 10(4) to 10(7) colony forming units/ml. S enteridis PT4 was also isolated from 8 of 20 fresh chilled chickens on retail sale. No other serotype of Salmonella or phage type of S enteritidis was cultured either from the chickens with pericarditis or from the fresh chilled chicken. PMID- 2569612 TI - The AIDS challenge. PMID- 2569613 TI - Cluster of childhood Guillain-Barre cases after an oral poliovaccine campaign. PMID- 2569614 TI - Mental test scores and adult education. PMID- 2569615 TI - Lutein, cholesterol, and risk of cancer. PMID- 2569616 TI - Copper deficiency in Wilson's disease. PMID- 2569617 TI - Aspirin, ticlopidine, and stroke. PMID- 2569618 TI - Dipyridamole. PMID- 2569619 TI - Fish oil and coronary angioplasty. PMID- 2569620 TI - Worldwide increase in pneumococcal antibiotic resistance. PMID- 2569621 TI - Erythromycin resistance in streptococci. PMID- 2569622 TI - Regression of cutaneous metastatic malignant melanoma with topical diphencyprone and oral cimetidine. PMID- 2569623 TI - Pancreatitis induced by 5-aminosalicylic acid. PMID- 2569625 TI - Sex ratio of infants following assisted reproduction. PMID- 2569624 TI - Naloxone hazard in drug abuser. PMID- 2569626 TI - Brain damage by neonatal hypoglycaemia. PMID- 2569627 TI - Trial abandoned, in disgust. PMID- 2569628 TI - Auditory risks in percutaneous lithotripsy. PMID- 2569629 TI - Cyclosporin metabolites and neurotoxicity. PMID- 2569630 TI - Epstein-Barr virus/complement receptor and epithelial cells. PMID- 2569632 TI - Transient cardiomegaly in tiny babies. PMID- 2569631 TI - Maternal serum specific beta 1-glycoprotein in pregnancies associated with Down's syndrome. PMID- 2569633 TI - Triazolam syndrome 10 years on. PMID- 2569634 TI - Galactose and ovarian cancer. PMID- 2569635 TI - Nutritional lessons from war-time Sierra Leone. PMID- 2569636 TI - Diabetes mellitus in Africans. PMID- 2569637 TI - Maternal and infant mortality. PMID- 2569638 TI - Occult cervical carcinoma revealed by large loop diathermy. PMID- 2569639 TI - Hypervariable microsatellite for genetic diagnosis. PMID- 2569640 TI - Maternal pyrexia and epidural anaesthesia. PMID- 2569641 TI - Seasonal clustering of sarcoidosis. PMID- 2569642 TI - Dermatofibromas and arthropod bites. PMID- 2569643 TI - Serological diagnosis of Salmonella infections. PMID- 2569644 TI - Agranulocytosis and thrombocytopenia in patient with Blackfan-Diamond anaemia during oral chelator trial. PMID- 2569645 TI - Safety of oral iron chelator L1. PMID- 2569647 TI - Behringwerke anti-HIV test kits. PMID- 2569646 TI - Pentamidine and hypoglycaemia. PMID- 2569648 TI - Anaphylactic reaction to injected hazelnut. PMID- 2569649 TI - HIV-associated immune thrombocytopenia. PMID- 2569650 TI - Women receiving anti Rh(D) immunoglobulin containing HIV antibodies. PMID- 2569651 TI - Duty to disclose medical records. PMID- 2569652 TI - AIDS surveillance. PMID- 2569653 TI - Blood pressure reduction in hypertensive-diabetic rats by the somatostatin analog MK-678. AB - A hypotensive effect of an orally-administered cyclopeptide somatostatin analog, MK-678, has been demonstrated in a hypertensive diabetic rat model. Sustained blood pressure reduction failed to occur when the drug was administered to the spontaneously hypertensive rat. The mechanism of hypotension appears independent of effects on a variety of hormones including insulin, glucagon, growth hormone, and components of the renin-angiotensin system including renin activity, plasma angiotensin converting enzyme, and aldosterone. PMID- 2569654 TI - Stereochemistry of the metabolism of MDMA to MDA. AB - The chiral derivatizing reagent N-trifluoroacetyl-L-prolyl chloride (LTPC) was used to form diastereomers of 3,4-methylenedioxymethamphetamine (MDMA) and 3,4 methylenedioxyamphetamine (MDA) which were resolved on an achiral gas chromatographic column using a mass spectrometer as a detector. Rats were subcutaneously dosed with 40 mg/kg of (+/-) MDMA.HCl and blood was obtained by decapitation four hours after dosing. Plasma was separated and extracted. The extract was derivatized on-column with LTPC. In addition to the two MDMA isomers, the demethylated metabolites, S(+) and R(-)-MDA were identified. In all experimental groups (male rats, food deprived male rats, female rats, post partum female rats, and mice) dosed with racemic MDMA, higher levels of the S(+) isomer of MDA relative to the R(-) MDA isomer were observed. This may be significant since it has been shown that the S(+) isomer of MDMA is the more neurotoxic isomer of the racemic drug of abuse MDMA. PMID- 2569655 TI - Relative efficacies of 1,4-diazepines on GABA-stimulated chloride influx in rat brain vesicles. AB - The effects of 1,4-diazepines with two annelated heterocycles [brotizolam (WE 941), ciclotizolam (WE 973) and WE 1008] on gamma-aminobutyric acid (GABA) stimulated chloride influx into rat brain membrane vesicles were examined. Brotizolam enhanced GABA (30 microM)-stimulated 36Cl- influx (146.1% of control), while ciclotizolam and WE 1008 showed only a small enhancement (119.3% and 119.1%, respectively) of GABA-stimulated 36Cl- uptake. Brotizolam resulted in a left shift of the GABA dose response curve at lower concentrations of GABA (10 microM), while at higher concentrations of GABA (1 mM), brotizolam caused a reduction of the maximal response. The enhancement of GABA-stimulated 36Cl- uptake by brotizolam (0.1 microM) was antagonized by Ro 15-1788. At higher concentration of GABA (300 microM), brotizolam inhibited GABA-stimulated 36Cl- uptake in a dose dependent manner and Ro15-1788 failed to antagonize this effect. These results suggest that 1) brotizolam produces an enhancement of GABA (30 microM)-stimulated chloride influx through the benzodiazepine receptor. 2) brotizolam inhibition of GABA (300 microM)-stimulated chloride influx involves an additional mechanism, and 3) the sedative-hypnotic action of brotizolam may be related to its high efficacy at the benzodiazepine/GABA-gated chloride channel. PMID- 2569657 TI - Isolation and identification of acetyl-CoA carboxylase from rainbow trout (Salmo gairdneri) liver. AB - Acetyl-CoA carboxylase is the pivotal enzyme in the de novo synthesis of fatty acids and is the only carboxylase with a biotin-containing subunit greater than 200,000 daltons. The biotin moiety is covalently linked to the active site and has a high affinity (Kd = 10(-15) M) for the protein avidin. This relationship has been used in previous studies to identify acetyl-CoA carboxylase isolated from mammalian species. However, acetyl-CoA carboxylase has not been isolated and characterized in a poikilothermic species such as the rainbow trout. The present study describes the isolation and identification of acetyl-CoA carboxylase in the cytosol of rainbow trout (Salmo gairdneri) liver. The enzyme was isolated using two distinct procedures--polyethylene glycol precipitation and avidin-Sepharose affinity chromatography. Identification of the isolated protein as acetyl-CoA carboxylase was made by the following: (1) sodium dodecyl sulfate-polyacrylamide gel electrophoresis; (2) avidin binding; (3) in vivo labeling with [14C]biotin; and (4) acetyl-CoA carboxylase-specific activity. The subunit molecular weight of the major protein was 230,000 daltons +/- 3.3%. This protein was shown to bind avidin (Mr = 16,600) prior to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, indicating the presence of biotin. In addition, protein isolated from fish that had previously received intraperitoneal injections of [14C]biotin, showed the majority of radioactivity associated with the 230,000 dalton protein.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569658 TI - Naloxone abolishes the inhibiting effect of somatostatin on the release of oxytocin evoked by insulin-induced hypoglycemia in humans. AB - The effect of somatostatin (SRIH) on the release of oxytocin (OT) in response to hypoglycemia during insulin tolerance test (ITT) was studied in seven normal men. Subjects were injected intravenously with 0.15 U/kg insulin alone (control test) or together with SRIH (4.1 micrograms/min x 90 min), naloxone (10 mg in an IV bolus), or the combination of the two substances. Plasma OT concentrations rose significantly during ITT; the OT response was significantly reduced by the treatment with SRIH and increased in the presence of naloxone. When both SRIH and naloxone were given, the OT response to hypoglycemia did not differ from that observed in the control test. These findings provide evidence that the effect of hypoglycemia on plasma OT levels is sensitive to the inhibition by SRIH and by naloxone-sensitive endogenous opioids. Because naloxone reversed the inhibiting effects of SRIH, an involvement of opioid peptides in SRIH action might be supposed. Alternatively, SRIH and naloxone-sensitive opioids might produce their inhibiting effects on OT rise in response to hypoglycemia through independent pathways. PMID- 2569656 TI - Incorporation of arachidonic, dihomogamma linolenic and eicosapentaenoic acids into cultured V79 cells. AB - The uptake and distribution of three common dietary polyunsaturated fatty acids was studied using Chinese hamster lung fibroblasts (V79 cells). Treatment of V79 cells with arachidonic (20:4), eicosapentaenoic (20:5) and dihomogammalinolenic (20.3) acids for 24 hr produced a marked uptake of 20:3 and 20:4, both of which were assimilated to a considerably greater degree than 20:5. All polyunsaturated fatty acids were incorporated primarily into phospholipids; however, there were considerable differences in their distribution into individual phospholipid species. Although 20:4 was incorporated primarily into phosphatidylcholine, 20:3 entered largely into phosphatidylethanolamine and phosphatidylglycerol, and 20:5 was distributed about equally between phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol. A marked conversion of 20:3 to 20:4 was found after 24 hr and, in several phospholipids, there was as much derived-radiolabeled 20:4 as there was radiolabeled 20:3. There was little evidence of 20:4 and 20:5 metabolism. V79 cells undergo substantial changes in phospholipid fatty acid composition following supplementation with these polyunsaturated fatty acids; however, these fatty acids are assimilated to different degrees and their distribution among cellular phospholipids is distinct, suggesting incorporation via independent mechanisms. PMID- 2569659 TI - Metabolism of 3H- and 14C-labeled glutamate, proline, and alanine in normal and adrenalectomized rats using different sites of tracer administration and sampling. AB - Alanine, glutamate and proline labeled with 14C and 3H were infused into fasted normal and adrenalectomized rats. Alanine was administered by the A-V mode (arterial administration-venous sampling), and glutamate and proline by both the A-V and V-A (venous administration-arterial sampling) modes. The kinetics of 14C alanine and 14C glutamate differed markedly from those of the tritium-labeled compounds, but there was little difference in the kinetics of 3H and 14C proline. The replacement rate calculated from the A-V mode for glutamate was about half that obtained in the V-A mode, but there was little difference with proline. The masses of the amino acids (total content of amino acids in the body) were calculated from the washout curves of the tritium-labeled compounds after the infusion of tracer was terminated. The masses for the normal rats were 407 mumol/kg for alanine, 578 mumol/kg for glutamate and 296 mumol/kg for proline. The so-called distribution spaces calculated conventionally from total masses and the amino acid concentrations in plasma are much greater than the volume of the body, reflecting the fact that amino acid concentrations in tissues greatly exceed those in plasma. Adrenalectomy markedly affected the kinetics of the three amino acids, and their replacement rates were greatly reduced. The proline and glutamate masses were reduced by at least one half, while that of alanine was unchanged. Adrenalectomy markedly reduced the conversion of proline to glutamate. The hydrocortisone regimen used in this study restored the metabolism of alanine and glutamate to normal, but had no effect on that of proline. PMID- 2569660 TI - The determination of mass of metabolites with tracers. AB - Application of tracers in vivo for the determination of replacement and mass of bloodborne compounds at steady state is discussed. Theory and methods to determine mass with tracers (total amount of compound-tracee-within the body) for compartmental and noncompartmental systems are presented, and their limitations examined. Methods to derive mass from the specific activity curves after bolus injection or infusion of tracer are described using graphic procedures or by equations using the parameters of exponential curves. The relationship between assumed models and the interpretation of tracer data is examined. The determination of both replacement (appearance, which equals utilization at steady state) and mass of most compounds present in both extracellular and intracellular fluids (such as lactate and amino acids) requires the application of the A-V mode for tracer administration and sampling of blood. Recycling of carbon affects the determination of mass with 14C. Estimates of true mass are provided with tritium labeled compounds, even when tritium loss is by exchange with protons or through futile cycling. Estimates of the amount (body mass) of lactate, alanine, glutamate, and proline obtained with tritium-labeled compounds are presented. Most of these masses are intracellular. The concentration of lactate in tissues equals or is greater, and that of amino acids much greater than that in plasma. Hence, the so-called "distribution space" for these compounds, calculated conventionally by dividing mass by plasma concentration, would appear to be equal to or greater than the body water of lactate, and several liters per kilogram for amino acids. PMID- 2569662 TI - Glutamine metabolism in the liver: overview and current concepts. PMID- 2569661 TI - Impairment of glutathione metabolism in erythrocytes from patients with diabetes mellitus. AB - The metabolism of glutathione and activities of its related enzymes were studied in erythrocytes from patients with non-insulin-dependent diabetes mellitus (NIDDM). A decrease in the levels of the reduced form of glutathione and an increase in the levels of glutathione disulfide were found in erythrocytes of diabetics. To elucidate these changes in the levels of glutathione, synthetic and degradative processes were studied. The activity of gamma-glutamylcysteine synthetase was significantly lower in diabetics than in normal controls. The activity of glutathione synthetase of each group was the same. The rate of outward transport of glutathione disulfide in diabetics decreased to approximately 70% of that of normal controls. The activity of glutathione reductase decreased in diabetics. These data suggest that the decrease in the levels of reduced form of glutathione in erythrocytes of diabetics is brought about by impaired glutathione synthesis and that the increase in the levels of glutathione disulfide is brought about by the decreased transport activity of glutathione disulfide through the erythrocyte membrane together with a decrease in the activity of glutathione reductase. These data also suggest that the impairment of glutathione metabolism weakens the defense mechanism against oxidative stress in erythrocytes of diabetics. PMID- 2569663 TI - Metabolism of glutamine in lymphocytes. AB - Pathways of glutamine metabolism in resting and proliferating rat thymocytes and established human T- and B-lymphoblastoid cell lines were evaluated by in vitro incubations of freshly prepared or cultured cells for one to two hours with [U14C]glutamine. Complete recovery of glutamine carbons utilized in products allowed quantification of the pathways of glutamine metabolism under the experimental conditions. Partial oxidation of glutamine via 2-oxoglutarate in a truncated citric acid cycle to CO2 and oxaloacetate, which then was converted to aspartate, accounted for 76% and 69%, respectively, of the glutamine metabolized beyond the stage of glutamate by resting and proliferating thymocytes. Similar results were obtained with the lymphoblastoid T- and B-cell lines. Complete oxidation to CO2 in the citric acid cycle via 2-oxoglutarate dehydrogenase and isocitrate dehydrogenase accounted for only 25% and 7%, respectively. In proliferating cells a substantial amount of glutamine carbons was also recovered in pyruvate, alanine, and especially lactate. The main route of glutamine and glutamate entrance into the citric acid cycle via 2-oxoglutarate in lymphocytes appears to be transamination by aspartate aminotransferase rather than oxidative deamination by glutamate dehydrogenase. In the presence of glucose as a second substrate, glutamine utilization and aspartate formation markedly decreased, but complete oxidation of glutamine carbons to CO2 increased to 37% and 23%, respectively, in resting and proliferating cells. The dipeptide, glycyl-L glutamine, which is more stable than free glutamine, can substitute for glutamine in thymocyte cultures at higher concentrations. PMID- 2569664 TI - Utilization of intravenously administered N-acetyl-L-glutamine in humans. AB - L-glutamine is too unstable for inclusion in solutions for parenteral nutrition, but its acetylated analogue, N-acetyl-L-glutamine is not. The purpose of this three-part study was to investigate the utilization of intravenously (IV) administered acetylglutamine in humans. In study 1, nine healthy postabsorptive subjects were given 9.4 g acetylglutamine IV during four hours. In study 2, five healthy subjects were studied on two occasions following an overnight fast. They were given 9.4 g of acetylglutamine or an equivalent amount of glutamine as part of a total parenteral nutrition (TPN) regimen during 7.2 hours. A control group of five subjects was given the same TPN regimen, but without acetylglutamine or glutamine. The nutrient solution included glucose, amino acids, and a fat emulsion, supplying 9.4 g nitrogen and 6,300 kJ in a total volume of 1.8 L. In study 3, four patients were studied the day after major surgery. They were given the same TPN regimen as in study 2, containing 9.4 g acetylglutamine, during 7.2 hours. Plasma concentrations and urinary excretion of acetylglutamine and glutamine were measured in all three studies, and so were splanchnic and renal exchange of acetylglutamine and glutamine in study 1. In study 1, the plasma concentration of glutamine rose from 594 +/- 28 mumol/L to 728 +/- 26 mumol/L (P less than .001), whereas plasma levels of acetylglutamine exceeded 1,000 mumol/L in all subjects at the end of infusion. The eight-hour urinary excretion of acetylglutamine and glutamine corresponded to 18% of the infused amount of acetylglutamine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569665 TI - [Double-blind study of clobazam (Odipam) and diazepam in non-psychotic anxiety states]. AB - Clobazam (Odipam) and Diazepam were investigated by the double blind study method in patients with nonpsychotic anxiety. It was found out that both drugs had equal anxiolytic and tranquilizing effect with the same adverse effects. Greater frequency of excellent marks concerning the final therapeutic effect of Odipam in respect to other marks pointed to its better clinical tolerance, so that this drug was recommended as daily tranquilizer. PMID- 2569666 TI - Glaxo presents its views ... PMID- 2569667 TI - Update: HIV-2 infection--United States. PMID- 2569668 TI - Organization and expression of the cell cycle gene, ts11, that encodes asparagine synthetase. AB - The human ts11 gene was isolated on the basis of its ability to complement the mutation of the BHK cell cycle ts11 mutant, which is blocked in G1 at the nonpermissive temperature. This gene has now been identified as the structural gene for asparagine synthetase (AS) on the bases of sequence homology and the ability of exogenous asparagine to bypass the ts11 block. The ts11 (AS) mRNA has a size of about 2 kilobases and is induced in mid-G1 phase in human, mouse, and hamster cell lines. We have studied the organization and regulation of expression of the ts11 gene. The human ts11 gene consists of 13 exons (the first two noncoding) interspersed in a region of about 21 kilobases of DNA. Transient expression assays using the bacterial chloramphenicol acetyltransferase reporter gene identified two separate promoters: one (ts11 P1) contained in a 280-base pair region upstream of the first exon and the other (ts11 P2) contained in the first intron. ts11 P1 produced about sixfold more chloramphenicol acetyltransferase activity than did ts11 P2 and had features of the promoters of housekeeping genes: high G + C content, multiple transcription start sites, absence of a TATA box, and presence of putative Sp1 binding sites. ts11 P2 contained a TATA sequence and other elements characteristic of a promoter, but so far we have no evidence of its physiological utilization. The ts11 gene was overexpressed in ts11 cells exposed to the nonpermissive temperature. Addition of asparagine to the culture medium led to a drastic decrease in mRNA levels and prevented G1 induction in serum-stimulated cells, which indicated that expression of the AS gene is regulated by a mechanism of end product inhibition. PMID- 2569670 TI - Suppression of experimental autoimmune encephalomyelitis by sulfasalazine. PMID- 2569669 TI - Evolution and stability of chromosomal DNA coamplified with the CAD gene. AB - We have compared clones of Syrian hamster cells selected for the first amplification of the CAD gene with clones selected for further amplification. The large domain amplified initially was not reamplified as an intact unit. Instead, subregions were reamplified preferentially, and parts of the initial array were often lost. These events reduced the average amount of coamplified DNA accompanying each copy of the selected gene. The degree of amplification was small in the first step (about three extra copies of CAD per cell), but second step amplifications to a high copy number (up to 60 extra copies per cell) occurred frequently. After several separate steps of amplification, highly condensed arrays that brought many CAD genes close together were formed. In striking contrast to the stability of these highly amplified arrays, the low-copy chromosomal arrays formed early were quite unstable and were often lost completely within 1 or 2 months of growth without selection. The results suggest that different mechanisms may be involved in the first step of amplification and in the later evolution of an already amplified array. PMID- 2569671 TI - Changes in the amounts of amino acids associated with nerve endings (synaptosomes) during synaptogenesis. AB - Changes in the amounts of proteins and amino acids in synaptosomes and whole tissue from the olfactory bulb and cerebral cortex of rats were measured during the period 5-25 days postnatal. The amount of neurotransmitter type amino acids (such as GABA, glutamate and aspartate) associated with synaptosomes obtained from 1g of brain tissue increased dramatically with the age of the animals, whereas non-transmitter type amino acids (such as serine and glutamine) showed relatively little change. The results were in harmony with an earlier cessation of synaptogenesis in the olfactory bulb than in the cerebral cortex. PMID- 2569672 TI - The immunolysis, isolation, and properties of subpopulations of mammalian brain synaptosomes. AB - Five subpopulations of mammalian brain synaptosomes can be selectively damaged by complement-mediated immunolysis employing antibodies to specific surface markers for each subpopulation. This allows the size of these subpopulations to be estimated. Employing antibodies alone, it has proved possible to isolate three of these subpopulations in very pure preparations which are metabolically viable. The immunoaffinity technique involved (immunomagnetophoresis) uses magnetic microspheres and produces mg (protein) quantities of synaptosomes. PMID- 2569673 TI - Inhibition by excitatory sulphur amino acids of the high-affinity L-glutamate transporter in synaptosomes and in primary cultures of cortical astrocytes and cerebellar neurons. AB - A detailed kinetic study of the inhibitory effects of L- and D-enantiomers of cysteate, cysteine sulphinate, homocysteine sulphinate, homocysteate, and S sulpho-cysteine on the neuronal, astroglial and synaptosomal high-affinity glutamate transport system was undertaken. D-[3H] Aspartate was used as the transport substrate. Kinetic characterisation of uptake in the absence of sulphur compounds confirmed the high-affinity nature of the transport systems, the Michaelis constant (Km) for D-aspartate uptake being 6 microM, 21 microM and 84 microM, respectively, in rat brain cortical synaptosomes and primary cultures of mouse cerebellar granule cells and cortical astrocytes. In those cases where significant effects could be demonstrated, the nature of the inhibition was competitive irrespective of the neuronal versus glial systems. The rank order of inhibition was essentially similar in synaptosomes, neurons and astrocytes. Potent inhibition (Ki approximately Km) of transport in each system was exhibited by L-cysteate, and L- and D-cysteine sulphinate whereas substantially weaker inhibitory effects (Ki greater than 10-1000 times the appropriate Km value) were exhibited by the remaining sulphur amino acids. In general, inhibition: (i) was markedly stereospecific in favor of the L-enantiomers (except for cysteine sulphinate) and (ii) was found to decrease with increasing chain length. Computer assisted molecular modelling studies, in which volume contour maps of the sulphur compounds were superimposed on those of D-aspartate and L-glutamate, demonstrated an order of inhibitory potency which was, qualitatively, in agreement with that obtained quantitatively by in vitro kinetic studies. PMID- 2569675 TI - Glutamine and 2-oxoglutarate as metabolic precursors of the transmitter pools of glutamate and GABA: correlation of regional uptake by rat brain synaptosomes. AB - To more clearly define the roles of glutamine and 2-oxoglutarate as metabolic precursors of the transmitter pools of glutamate and GABA we have determined the relative rates at which these four substances, and adenosine and serotonin are accumulated by synaptosomes derived from twelve regions of the rat brain. Initial transport conditions and low substrate concentrations were used to maximize uptake by high-affinity systems, except the uptake of glutamine was determined at both low and high concentrations. Because the uptake of 2-oxoglutarate is markedly enhanced by glutamine, 2-oxoglutarate uptake was determined with and without glutamine (0.2 mM) added to the incubation medium. For each substrate, regional differences in uptake ranged from approximately two- to fourteen-fold. An anaylsis of uptake kinetics revealed that the regional differences were due primarily to differences in transport capacity rather than substrate affinities, at least for glutamate, GABA, and 2-oxoglutarate. Thirty-four correlation analyses of relative uptake values were performed. Strong correlations were found between 2-oxoglutarate and glutamate, and between glutamine and glutamate, whereas no strong correlations occurred between these substrates and GABA. Our results support the view that both glutamine and 2-oxoglutarate are major precursors of the transmitter pool of glutamate throughout the rat brain, but their relative contributions toward replenishing the transmitter pool of GABA are less certain. PMID- 2569674 TI - Role of aspartate aminotransferase and mitochondrial dicarboxylate transport for release of endogenously and exogenously supplied neurotransmitter in glutamatergic neurons. AB - Evoked release of glutamate and aspartate from cultured cerebellar granule cells was studied after preincubation of the cells in tissue culture medium with glucose (6.5 mM), glutamine (1.0 mM), D[3H] aspartate and in some cases aminooxyacetate (5.0 mM) or phenylsuccinate (5.0 mM). The release of endogenous amino acids and of D-[3H] aspartate was measured under physiological and depolarizing (56 mM KCl) conditions both in the presence and absence of calcium (1.0 mM), glutamine (1.0 mM), aminooxyacetate (5.0 mM) and phenylsuccinate (5.0 mM). The cellular content of glutamate and aspartate was also determined. Of the endogenous amino acids only glutamate was released in a transmitter fashion and newly synthesized glutamate was released preferentially to exogenously supplied D [3H] aspartate, a marker for exogenous glutamate. Evoked release of endogenous glutamate was reduced or completely abolished by respectively, aminooxyacetate and phenylsuccinate. In contrast, the release of D-[3H] aspartate was increased reflecting an unaffected release of exogenous glutamate and an increased "psuedospecific radioactivity" of the glutamate transmitter pool. Since aminooxyacetate and phenylsuccinate inhibit respectively aspartate aminotransferase and mitochondrial keto-dicarboxylic acid transport it is concluded that replenishment of the glutamate transmitter pool from glutamine, formed in the mitochondrial compartment by the action of glutaminase requires the simultaneous operation of mitochondrial keto-dicarboxylic acid transport and aspartate aminotransferase which is localized both intra- and extra mitochondrially. The purpose of the latter enzyme apparently is to catalyze both intra- and extra-mitochondrial transamination of alpha-ketoglutarate which is formed intramitochondrially from the glutamate carbon skeleton and transferred across the mitochondrial membrane to the cytosol where transmitter glutamate is formed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569677 TI - Effects of subchronic nicotine administration on central dopaminergic mechanisms in the rat. AB - Nicotine was administered acutely and subchronically (14 days) to determine whether various synaptic mechanisms are selectively altered in the nigrostriatal and mesolimbic dopaminergic systems in the rat. When added to tissue preparations in vitro, nicotine had no effects on tyrosine hydroxylase, synaptosomal uptake of [3H]dopamine or binding of [3H]spiperone to D2 receptors in either system. However, acute treatment in vivo stimulated tyrosine hydroxylase activity in the nucleus accumbens. This effect was prevented by pretreatment with a nicotinic antagonist, suggesting that it was mediated by nicotinic receptors. Since subchronic exposure to nicotine had no effect on tyrosine hydroxylase, it appears that tolerance develops to this action. In vivo treatment with nicotine did not alter dopamine uptake or receptor binding. The results suggest that, in doses which result in moderate plasma levels, nicotine has selective stimulant actions on nerve terminals of the mesolimbic system. PMID- 2569676 TI - Differential effects of ammonia and beta-methylene-DL-aspartate on metabolism of glutamate and related amino acids by astrocytes and neurons in primary culture. AB - The effects of ammonium chloride (3 mM) and beta-methylene-DL-aspartate (BMA; 5 mM) (an inhibitor of aspartate aminotransferase, a key enzyme of the malate aspartate shuttle (MAS] on the metabolism of glutamate and related amino acids were studied in primary cultures of astrocytes and neurons. Both ammonia and BMA inhibited 14CO2 production from [U-14C]- and [1-14C]glutamate by astrocytes and neurons and their effects were partially additive. Acute treatment of astrocytes with ammonia (but not BMA) increased astrocytic glutamine. Acute treatment of astrocytes with ammonia or BMA decreased astrocytic glutamate and aspartate (both are key components of the MAS). Acute treatment of neurons with ammonia decreased neuronal aspartate and glutamine and did not apparently affect the efflux of aspartate from neurons. However, acute BMA treatment of neurons led to decreased neuronal glutamate and glutamine and apparently reduced the efflux of aspartate and glutamine from neurons. The data are consistent with the notion that both ammonia and BMA may inhibit the MAS although BMA may also directly inhibit cellular glutamate uptake. Additionally, these results also suggest that ammonia and BMA exert differential effects on astroglial and neuronal glutamate metabolism. PMID- 2569678 TI - Regulation of glutamine synthetase synthesis and activity by glucocorticoids and adrenoceptor activation in astroglial cells. AB - Glucocorticoids are known to induce the synthesis and activity of glutamine synthetase (GS; EC 6.3.1.2.) in astroglial cells. In the present paper, noradrenaline (NA), in itself ineffective upon GS regulation, potentiated GS activity in astroglial primary cultures in the presence of the glucocorticoid dexamethasone, the GS activity being further stimulated in the presence of glutamate (glu). Thus, adrenoceptor activation might interact with the glucocorticoid induced GS activity in astroglial primary cultures. PMID- 2569680 TI - Effect of midbrain raphe nucleus stimulation on somatosensory evoked potential in cat. AB - Experiments were carried out to clarify the effect of serotonin [5 hydroxytryptamine (5-HT)] upon the somatosensory evoked potential (SEP) elicited by superficial radial nerve stimulation in cat, and the following results were obtained: (i) in untreated (control) animals, a significant reduction occurred in amplitude of the SEP in response to conditioning stimulation of the dorsal raphe nucleus in the midbrain, within 100 ms of the interval between conditioning and test stimuli; (ii) in 5-HT depleted animals prepared by intraperitoneal injection of p-chlorophenylalanine (PCPA), the amplitude of SEP significantly increased in comparison with that of nontreated animals. Conditioning stimulation of the raphe nucleus in 5-HT depleted animals failed to cause any remarkable change in the SEP; (iii) the diminuation of the SEP by raphe nucleus conditioning reappeared following intraventricular administration of a large dose of 5-HT (200 micrograms) in the PCPA-treated animals. These results have demonstrated that the serotonergic system with its cell bodies located in the raphe nucleus of the midbrain has the function of reducing the amplitude of the SEP, thus suggesting that it acts as a suppressive control on the excitability of the cortex to sensory input. PMID- 2569679 TI - The role of endothelin in the pathogenesis of vasospasm following subarachnoid haemorrhage. AB - The purpose of the present study was to evaluate the vasocontractile activity of endothelin, a newly isolated endothelium-derived constrictor peptide, in canine basilar arteries in vitro and in vivo. Endothelin at concentrations of 10(-12) M approximately 3 X 10(-8) M elicited dose-dependent contractions of canine basilar arteries in vitro. The maximum tension was larger than that induced by 40 mM KCl. The EC50 value was 1.9 +/- 0.6 X 10(-9) M (mean +/- SEM). The endothelin-induced contraction was reversed by 10(-8) M nicardipine or 10(-5) M approximately 10(-4) M papaverine. An intracisternal injection of 0.6 approximately 1.2 X 10(-12) mol/kg of endothelin caused biphasic contraction of the basilar artery lasting for more than 24 h. The initial phase of the contraction accompanied remarkable changes in vital signs such as an acute rise of blood pressure, bradycardia and respiratory arrest. An intracisternal injection of 2.0 X 10(-12) mol/kg of endothelin also induced acute contraction of the basilar artery. However, all of the dogs which received an intracisternal injection of 2.0 X 10(-12) mol/kg of endothelin died from sustained respiratory insufficiency. The present results demonstrate that endothelin induces strong and long-lasting contractions of cerebral arteries. Therefore, endothelin may play an important role in the pathogenesis of vasospasm. PMID- 2569681 TI - Intra-aortic balloon counterpulsation: a treatment for ischaemic stroke? AB - Intra-aortic balloon counterpulsation (IABC) augments cardiac output (CO) and pulse pressure (PP) allowing patients with low output heart failure to be supported for a period of time. Augmentation of CO and PP may also be beneficial to the patient with acute cerebral ischaemia. In this paper we investigated the possibility of using IABC to increase local cerebral blood flow (CBF) in ischaemic brain. In 12 anaesthetized mongrel dogs, a canine stroke model was produced by occluding the left internal carotid and middle cerebral arteries with aneurysm clips. Six dogs were then treated with IABC for 2 h, and 6 other dogs acted as controls (no IABC). Haemodynamic data were measured continuously and CBF (microsphere technique) and CO measurements were performed pre- and post occlusion, and then twice during the treatment period. In the IABC-treated animals, PP increased from 32 +/- 5.9 to 39 +/- 7.8 mmHg (p less than 0.01) but CO and local CBF in the ischaemic brain did not change significantly during IABC. However, in 4 dogs with significant increases in CO due to IABC [1.7 +/- 0.3 to 2.8 +/- 0.7 l/min (p less than 0.05)], local CBF in ischaemic brain also increased significantly from 22 +/- 12 to 26 +/- 11 cc/100 g/min (p less than 0.05). In the control animals, CO and local CBF did not change significantly during the observation period. These data suggest that augmentation of CO and PP by IABC results in an increase in local CBF in ischaemic brain. IABC may be an effective treatment for ischaemic stroke in those patients with compromised cardiac performance whose cardiac output and pulse pressure can be augmented by IABC. PMID- 2569682 TI - Dynamics of ischaemic brain oedema fluid: 'dense tracer distribution around the focus', evaluated by peroxidase injection in the rat cortex. AB - Experimental cerebral ischaemia was produced by microembolization in rats. HRP (horseradish peroxidase) was then injected into the cerebral cortex and the extracellular fluid dynamics were observed. In the control group, there was a basic pattern in the migration of the tracer. In the ischaemic group, a 'dense tracer distribution around the focus' was found in addition to the basic pattern. This phenomenon could be observed from the second day after the ischaemic insults. The perivascular DAB reaction was marked, particularly around the focus. There was a vesicular uptake of the tracer in the vascular endothelium. This phenomenon appeared almost simultaneously with the active resolution of the oedema fluid. In addition, frequent mitotic figures were seen around the focus. It was thus possible to hypothesize that the 'dense tracer distribution around the focus' appeared at the site of active tissue reaction and had a close relationship with oedema resolution. PMID- 2569683 TI - So-called intracranial germ cell tumours: personal experiences and a theory of their pathogenesis. AB - We investigated 110 cases of intracranial germ cell tumours (up to the end of 1986), 56% of which showed monotypic histological patterns and 44% were shown to be mixed tumours. All these cases underwent surgery followed by radiation and/or chemotherapy. All cases of choriocarcinoma and embryonal carcinoma died within 2 yr: cases of endodermal sinus tumour also showed poor results (4 yr survival rate was 12.5%). Mature teratoma had a 5 yr survival rate (5YSR) and a 10 yr survival rate (10YSR) of 78.3% each: immature teratoma showed a 5YSR of 44.9%. Two-cell pattern tumours (PTC) showed a 5YSR of 85.8% and a 10YSR of 82.4%. They (PTC) can be divided into two groups (i) germinoma and (ii) pinealoma of pineal parenchyma origin (pineocytoma with lymphocytic infiltration) on the basis of the difference in the tumour cell-stroma relationship and also placental alkaline phosphatase stain. In the pineal region, 70% of PTC belonged to the category of pinealoma and in the suprasellar region, 92% of PTC were germinoma. Germinoma showed a 5YSR of 91.4% and a 10YSR of 91.4%, whereas those of the pinealoma were 78.2% and 68.4% respectively. This suggests that the biological characteristics of germinoma and pinealoma may be different. All these results may bring into question the validity of the germ cell theory, since germinoma, which should be the most undifferentiated according to the theory, was the most benign and choriocarcinoma and endodermal sinus tumour, which should be the most differentiated, were the most malignant in the follow-up study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569684 TI - Role of cerebral endothelium in brain oedema. AB - The view that cerebral endothelial cells represent the cellular basis of the blood-brain barrier has been generally accepted. The regulation of transport processes operating in the cerebral endothelial cells is currently of great interest. Our knowledge of these regulatory mechanisms is briefly reviewed here with special regard to the molecular processes involved in the formation of brain oedema and emphasis on new therapeutic means for its prevention. PMID- 2569685 TI - Variations in red blood cell polyamines in brain tumour patients treated with stereotactic radiosurgery. AB - Red blood cell polyamine levels were followed in 13 patients treated with stereotactic radiosurgery for various types of brain tumours. In the most malignant tumours radiosurgery resulted in a decrease in spermine and an increase in spermidine levels. This discrepancy between variations in polyamine levels was not found after conventional radiotherapy. The results may indicate a change in tumour polyamine activity induced by high energy radiotherapy. PMID- 2569687 TI - Technical aids for the flexible use of the Leksell stereotactic system. AB - As part of a multimodality therapy program for intracranial tumours, 105 stereotactic and implant procedures have been carried out utilizing the CT compatible Leksell stereotactic system. In the iridium implant series, 86 catheters have been implanted for an average of 3.6 targets per patient. There have been no deaths or missed targets and only two incidential haemorrhages detected. In order to facilitate the reliability, safety and speed of multiple catheter insertion, several techniques have been developed including: (a) a standardized single-length catheter and flange system; (b) a ceramic catheter for microwave hyperthermia; (c) a mnemonic card for ease of calculation; (d) a radiation shield for nursing; (e) a stereotactic drill and surgical approaches to far lateral and posterior fossa targets. Principles for the use of these technical aids are discussed. PMID- 2569686 TI - Clinical study of OKY-046, a thromboxane synthetase inhibitor, in prevention of cerebral vasospasms and delayed cerebral ischaemic symptoms after subarachnoid haemorrhage due to aneurysmal rupture: a randomized double-blind study. AB - A double-blind study was conducted at 48 neurosurgical services in Japan to investigate the usefulness of OKY-046, an imidazole derivative and a thromboxane synthetase inhibitor, on cerebral vasospasm and cerebral ischaemic symptoms in patients with ruptured cerebral aneurysms. OKY-046 was administered in two daily doses of 80 mg (L group) and 400 mg (H group), and compared with a group given a placebo (P group). The following results were obtained: the occurrence of cerebral vasospasm was significantly lower in the L group than in the P group; the development of low density area (LD) in CTs was significantly lower in both the L and H groups than in the P group; motor paralysis in the L group improved significantly sooner, and that in the H group tended to improve sooner than that in the P group; in subjects with severe vasospasm, the incidence of LD was significantly lower, with better functional prognosis, in the L group than in the P group; in subjects with severe grades on the Glasgow Coma Scale (GCS), Japan Coma Scale (JCS) or High Density (HD) Score the functional prognosis at 1 month after the aneurysmal rupture was significantly better in the L group than in the P group, though no significant differences were seen in the overall investigation; there were no significant differences among the three groups in the development of either laboratory-determined abnormality or of adverse reactions. It is thus concluded that OKY-046 is clinically useful at a dose of 80 mg/d for cerebral vasospasm and cerebral ischaemic symptoms after subarachnoid haemorrhage (SAH) caused by aneurysmal rupture. PMID- 2569688 TI - The effect of locus ceruleus lesion on water, sodium and potassium content of the rat cerebral cortex. AB - Locus ceruleus lesion decreases the density of ouabain binding sites, and presumably Na+, K+-ATPase, in brain microvessels. To determine if this decrease affects the transport of Na+, K+ or water across the blood-brain barrier, we studied the influence of unilateral locus ceruleus lesion on Na+, K+ and water content of the ipsilateral cerebral cortex. Unilateral locus ceruleus lesion depleted norepinephrine in the ipsilateral cerebral cortex but had no effect on tissue Na+, K+ or water under steady-state conditions. When the Na+/K+ exchange pump of the blood-brain barrier was stressed by hyperkalaemia, K+ content in the ipsilateral cerebral cortex rose to higher levels than in the contralateral cortex, but the difference did not reach statistical significance. Locus ceruleus lesion also did not cause significant differences in the cerebral cortical content of water, Na+ or K+ in hyponatraemia. The results suggest that brain water and ion homeostasis are tightly controlled, probably by multiple mechanisms with biological redundancies, so that even a 50% decrease in the density of ouabain binding sites in brain endothelium does not result in significant alterations in brain water, Na+ or K+ content. PMID- 2569689 TI - Blockade of alpha 2-adrenergic receptors, but not blockade of gamma-aminobutyric acidA, serotonin, or opiate receptors, augments responsiveness of locus coeruleus neurons to excitatory stimulation. AB - Previous studies in this laboratory indicated that alpha 2-adrenergic receptors in the locus coeruleus play a major role in regulating the responsiveness of neurons in the locus coeruleus to excitatory influences. The present study points to the possibility that alpha 2-receptors are unique among inhibitory receptors in the locus coeruleus in regulating responsiveness of these neurons independently of the spontaneous firing rate. In the first part of the study, blockade of alpha 2-receptors was shown to markedly augment responsiveness of neurons in the locus coeruleus to the excitatory stimulus of compression of the contralateral hind paw at doses of an alpha 2-blocker both above and well below those necessary to increase spontaneous activity of neurons in the locus coeruleus. In contrast, blockade of gamma-aminobutyric acid and serotonin receptors augmented spontaneous firing rates of neurons in the locus coeruleus but failed to augment responsiveness of these neurons to compression of the hindpaw. Blockade of opiate receptors failed to increase either spontaneous firing rates or the responsiveness of neurons of the locus coeruleus to paw compression; moreover, in animals given an opiate agonist over a number of days to produce tonic stimulation of opiate receptors, blockade of opiate receptors augmented spontaneous firing rates of neurons in the locus coeruleus but had no effect on responsiveness to paw compression. In that blockade of each type of inhibitory receptor tested increased the spontaneous firing rates of neurons in the locus coeruleus but only blockade of alpha 2-receptors increased the responsiveness of neurons in the locus coeruleus to stimulation, without affecting the spontaneous firing rate, alpha 2-receptors may be unique among inhibitory receptors in independently regulating the responsiveness of neurons in the locus coeruleus. One possibility discussed for why alpha 2-receptors regulate the responsiveness, independently of the spontaneous firing rate, is that there are two types of alpha 2-receptors in the locus coeruleus, one of which regulates responsiveness and another which regulates the spontaneous firing rate. PMID- 2569690 TI - Comparison of opioid and GABA receptor control of excitability and membrane conductance in hippocampal CA1 pyramidal cells in rat. AB - Opioids are thought to increase the excitability of hippocampal pyramidal cells by decreasing release of neurotransmitter from inhibitory interneurons. This study compared the actions of the opioid agonist normorphine, and the GABA receptor antagonist bicuculline, on the responses of CA1 pyramidal cells to afferent stimulation. Both normorphine and bicuculline increased the sensitivity of pyramidal cells to presynaptic stimulation, increased the number of population spikes and action potentials elicited, increased the duration of the excitatory postsynaptic potential (EPSP) and reduced the change in input conductance during the early inhibitory postsynaptic potential (IPSP). Unlike bicuculline, normorphine also decreased the change in conductance during the late inhibitory postsynaptic potential. The decreased change in the conductance of pyramidal cells caused by normorphine during both early and late inhibitory postsynaptic potentials supports the hypothesis that opioids decrease the release of GABA from inhibitory interneurons. In addition to reducing GABA-mediated changes in conductance, both normorphine and bicuculline unmasked a D-APV-sensitive conductance, measured during the early inhibitory postsynaptic potential. These results demonstrate that activation of opioid receptors enhances the excitability of CA1 pyramidal cells by decreasing GABA-mediated early and late inhibitory postsynaptic potentials and by unmasking NMDA receptors. PMID- 2569691 TI - Binding sites for [3H]2-oxo-quazepam in the brain of the cat: evidence for heterogeneity of benzodiazepine recognition sites. AB - In the present study, the distribution of benzodiazepine recognition site subtypes in the brain of the cat was investigated. To this aim, the binding properties of [3H]2-oxo-quazepam ([3H]2OXOQ) and [3H]beta-CCE, two ligands with preferential affinity for Type I benzodiazepine recognition sites, were compared to binding parameters for [3H]flunitrazepam ([3H]FNT) in different areas of the cat brain. The ratio of [3H]2OXOQ to [3H]FNT binding sites indicated that, in the cerebellum, Type I sites accounted for 90% of the total number of benzodiazepine recognition sites. The cerebral cortex, thalamus and mesencephalic reticular formation had also a high proportion of Type I sites (73-78%), whilst the two subtypes were almost equally distributed in the hippocampus, amygdala and bulbar reticular formation. A similar distribution of subtypes of benzodiazepine recognition sites was indicated by the ratio of [3H]beta CCE to [3H]FNT binding sites for different areas of the brain. These results demonstrate the existence of heterogeneity of recognition sites for benzodiazepines in the brain of the cat and support the view that [3H]2OXOQ preferentially labels Type I sites. PMID- 2569692 TI - Blockade of histamine-stimulated alterations in cerebrovascular permeability by the H2-receptor antagonist cimetidine. AB - Histamine has been shown previously to cause dose-dependent systemic hypotension and concurrent alterations in the permeability of the blood-brain barrier of rats. The purpose of the present study was to determine whether histamine-induced changes in cerebrovascular permeability were mediated by the histamine H2 receptor. Wistar-Kyoto (control) and spontaneously hypertensive rats were pretreated with the histamine H2-receptor antagonist cimetidine (10 mg/kg), followed by saline or histamine (1.25, 2.5 or 5.0 micrograms/kg). Premedication with cimetidine did not block histamine-induced systemic hypotension. The permeability of the blood-brain barrier was measured with 131I-labelled serum albumin (RISA) or with 99mTc-sodium pertechnetate (TcO4-). In both control and spontaneously hypertensive animals, cimetidine prevented histamine-induced changes in the permeability of the blood-brain barrier to either tracer. These findings suggest that the H2-receptor is the prime mediator of histamine stimulated alterations in cerebrovascular permeability. PMID- 2569693 TI - Electroencephalographic changes and other indices of neurotoxicity with haloperidol-lithium therapy. AB - This prospective study was undertaken in order to establish indices of possible neurotoxicity due to the combination of lithium with haloperidol and a combination of lithium with other neuroleptics. Twenty-one subjects who were receiving neuroleptic-lithium combination were studied. Of them, 14 had a primary affective disorder-manic phase, and 7 had schizophrenia as judged by Feighner criteria. The subjects were evaluated on clinical, neuropsychological measures and EEG at baseline and 10-14 days after addition of lithium to the neuroleptic regimen. Of 11 subjects receiving the haloperidol-lithium combination, 5 (45.4%) showed abnormal responses on EEG. Of these 5 subjects, 3 showed abnormal photic responses, and 2 showed slowing in posterior alpha activity. These photoparoxysmal and photomyoclonic responses are indicative of cerebral pathophysiology resulting from the interactive effect of the haloperidol-lithium combination. The photic and other associated EEG abnormalities reported may be the earliest indication of neurotoxicity. No significant changes were observed in the EEG in 10 subjects treated with other neuroleptic-lithium combinations. In a statistical comparison the haloperidol-lithium combination had significantly more frequent EEG changes than the combination of lithium with other neuroleptics. This study presents sufficient evidence from reported photic and other associated EEG changes to pursue further investigation of neurotoxicity due to haloperidol lithium therapy. Such a study should employ a larger number of subjects, random assignment of subjects to treatment and control groups, and blind evaluation of data. PMID- 2569694 TI - Differential effects of pallidal lesions on the behavioural responses to SKF 38393, LY 171555 and apomorphine in the rat. AB - The purpose of this study was to determine the role of the globus pallidus in the expression of dopamine D1- and D2-receptor mediated motor events. Rats were first injected stereotaxically with 6-hydroxydopamine in one medial forebrain bundle to denervate the ascending dopamine pathways in that hemisphere. Apomorphine and selective D1 and D2 agonists were then administered, at two dose levels, to establish characteristic response patterns. Subsequently the animals were given a secondary lesion by injecting kainic acid (0.2-1 microgram) into the ipsilateral globus pallidus and retested with the dopamine agonists over a period of two months. The kainate treatment itself caused spontaneous motor asymmetries, followed by aphagia, adipsia and hypersensitivity to touch. Contraversive circling, contralateral posture and grooming induced by systemic apomorphine were all abolished by the kainate treatment, whilst sniffing and head movements were facilitated. All activities induced by D1 stimulation were abolished or severely reduced under these conditions. By contrast, the contralateral posture and grooming elicited by D2 stimulation were spared, and only D2-dependent contraversive rotation, sniffing and head movements were reduced. All behavioural deficits were temporary and recovered partially or completely during the course of the experiment, but could not be overcome by increasing the dose of dopamine agonist. Post mortem histology revealed a consistent loss of pallidal neurons, together with more variable damage to extrapyramidal structures and the thalamus. The results show that all the D1-mediated, and certain of the D2-mediated motor responses depend on the integrity of the pallidum for their expression in the unilaterally 6-hydroxydopamine-treated rat. PMID- 2569695 TI - Histochemical localization of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine oxidation in the mouse brain. AB - The sites in the mouse brain where 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine can be oxidized to the toxic metabolite 1-methyl-4-phenylpyridine were determined using a histochemical technique. The method involved the demonstration of monoamine oxidase activity using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine as the substrate by means of a sensitive coupled peroxidase technique. The distribution of neurons displaying the ability to oxidize 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine via a monoamine oxidase catalysed reaction was compared to that of various amine systems identified with immunohistochemistry. Dopamine neurons, and in particular the nigrostriatal dopamine cells, did not display the capacity to oxidize 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Noradrenergic neurons showed intense monoamine oxidase activity when 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine was used as substrate, and this activity was blocked by the monoamine oxidase-A inhibitor clorgyline. Serotonin neurons and histamine neurons were also able to oxidize 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine. The reaction in these neurons was blocked by deprenyl, an inhibitor of monoamine oxidase-B. Pretreatment with inhibitors of monoamine oxidase-B has been previously shown to prevent the neurotoxic action of 1-methyl 4-phenyl-1,2,3,6-tetrahydropyridine on dopaminergic neurons. Therefore, since serotonin and histamine neurons are able to oxidize 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine by means of monoamine oxidase-B, these neurons may be involved in the production of the toxic metabolites of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine in vivo. PMID- 2569696 TI - Effects of somatostatin-14 on the in vitro release of [3H]GABA from slices of rat caudatoputamen. AB - Slices (300 microns) of rat caudatoputamen were incubated in Krebs-Henseleit medium and loaded with [3H]glutamine, part of which was converted to [3H]GABA. This conversion takes place only in GABA-neurons most of which probably contribute to the striatonigral pathway. After a 24 min equilibration period, release of radioactivity was stimulated with veratridine (3.1-4 mumol/l) or K+ (15-25 mmol/l) in the absence or presence of somatostatin-14. From the radioactivity released [3H]GABA was separated by cationic exchange chromatography and measured. Somatostatin-14 affected the release of [3H]GABA in a manner which depended on its concentration as well as on the extent of stimulus-evoked release. Somatostatin-14 (1 nmol/l) enhanced the moderate release (2-4% of tissue content) elicited by veratridine (3.1 mumol/l) or K+ (20 mmol/l), but had no effect on the more pronounced release (5-8% of tissue content) elicited by veratridine (4 mumol/l) or K+ (25 mmol/l). Somatostatin-14 (10 nmol/l) had no effect on the moderate release of [3H]GABA, but diminished the pronounced one. Further experiments provided evidence that the somatostatin-14-induced enhancement was not brought about by a direct action on GABA-neurons but was probably indirect, i.e. mediated by other striatal neurons. In contrast, the diminution of the release of [3H]GABA caused by somatostatin-14 may be due to its direct action on releasing neurons. Two antisera against somatostatin lowered the pronounced release indicating that endogenous somatostatin may also enhance the release of [3H]GABA. In addition, endogenous somatostatin seems also to be able to diminish the release under certain experimental conditions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569697 TI - Physiology and pharmacology of dopamine D2-receptors: their implications in dopamine-substitute therapy for Parkinson's disease. PMID- 2569698 TI - [Effect of somatostatin and salmon calcitonin on carcinoid syndrome. A clinical case]. AB - In carcinoid syndrome, calcitonin administration proved to have an effective action in reducing symptomatology. During the perioperative period, treatment with somatostatin, thanks to the ease of administration of the drug and the reductive action of gastroenteric secretion, demonstrated its usefulness in countering the action of serotonin. In the present case, the data reported in the most recent literature on the use of the two drugs in the carcinoid syndrome are confirmed. PMID- 2569699 TI - [Use of somatostatin in the treatment of gastro-duodenal hemorrhage]. AB - The results obtained in the last 4 years in the treatment of superior digestive haemorrhage are reported. Of 71 patients examined, haemorrhage control with i.v. somatostatin was obtained in 95% of cases (68 patients). Arrest of haemorrhage was temporary in 2 patients, permitting surgery of choice. Three patients failed to respond to medical therapy and were operated in emergency. One of these patients died. No unwanted side-effects were observed. PMID- 2569700 TI - Molecular target size of an L-glutamate receptor from the cockroach central nervous system. AB - L-Glutamate binding to cockroach central nervous system (CNS) membranes is unaffected by freeze-drying and subsequent rehydration. The dissociation constant is similarly unaltered by irradiation at a dose of 3 Mrad which affects only the number of biologically active binding sites. A molecular weight estimate for this insect CNS L-glutamate receptor was performed using irradiation inactivation analysis. By this method a molecular target size of approximately 77.8 kDa was determined. This figure is in agreement with the molecular weight found by other authors for L-glutamate binding sites in the rat brain. PMID- 2569701 TI - Primary afferent depolarization in the rat spinal cord is mediated by pathways utilising NMDA and non-NMDA receptors. AB - In the present experiments the dorsal root-evoked dorsal root potential (DR-DRP) has been measured in vitro from a mature rat sacrococcygeal preparation. The DR DRP is an index of presynaptic inhibition since it represents the depolarization of primary afferent terminals by gamma-aminobutyric acid (GABA) released synaptically from interneurones. The present study shows that the synaptic excitation of the GABAergic interneurons contains a large component resistant to the selective N-methyl-D-aspartate (NMDA) receptor antagonists 2-amino-5 phosphonopentanoate (AP5) (100 microM) and 3((+)-2-carboxypiperazin-4-yl)propyl-1 phosphonate (CPP) 20-100 microM. This non-NMDA receptor mediated component reflected in the DR-DRP was depressed markedly by the non-selective excitatory amino acid receptor antagonists kynurenate (1-2 mM) and 6-cyano-2,3-dihydroxy-7 nitro-quinoxaline (CNQX) (10-20 microM). Because previous reports show non cholinergic activation of Renshaw cells to be blocked by NMDA receptor antagonists the present observations suggest that pre- and postsynaptic inhibition in the spinal cord are mediated by different types of excitatory amino acid receptor. PMID- 2569702 TI - Glycine potentiates NMDA responses in rat hippocampal CA1 neurons. AB - When superfused onto rat hippocampal slices, glycine (0.1-0.5 mM) potentiated the depolarization induced by pressure application of NMDA in normal Krebs solution and the synaptic discharge evoked by stimulation of the Schaffer collateral commissural inputs to the CA1 pyramidal neurons bathed in Mg2+-free media; the effects were not prevented by strychnine. In addition, glycine partially reversed the blocking effect of D-2-amino-5-phosphonovalerate (AP5) on N-methyl-D aspartate (NMDA)-induced depolarization. These results show that glycine at relatively high concentrations potentiates the NMDA-mediated response in hippocampal slices. PMID- 2569703 TI - Coexistence of GABA and peptide immunoreactivity in non-pyramidal neurons of the basolateral amygdala. AB - Colocalization of gamma-aminobutyric acid (GABA) immunoreactivity with somatostatin (SOM), neuropeptide Y (NPY), cholecystokinin (CCK), and vasoactive intestinal polypeptide (VIP) immunoreactivity was demonstrated in non-pyramidal neurons of the basolateral amygdala using a two-color immunoperoxidase procedure. Approximately 80-90% of SOM- and NPY-positive neurons in the basolateral amygdala were also immunoreactive for GABA. Virtually all large CCK-positive neurons also exhibited GABA-like immunoreactivity. About one-half of VIP-positive neurons and small CCK-positive cells were also immunoreactive for GABA. PMID- 2569704 TI - Ecstasy abuse. PMID- 2569705 TI - Accurate prenatal diagnosis with novel polymerase chain reaction primers in a family with sporadic hemophilia A. AB - Novel oligonucleotide primers are described for the convenient and internally controlled detection of the BclI factor VIII polymorphism on agarose gels. The primers were used for prenatal diagnosis in a family in which only one individual was affected with hemophilia A. In such families, it is important for the mother to understand that uncertainty about the point of origin of the mutation precludes the accurate diagnosis of affected fetuses but does allow the accurate diagnosis of unaffected fetuses in many cases. PMID- 2569706 TI - Calcium hydroxide in the management of traumatized permanent teeth. PMID- 2569707 TI - The Ad5 12S growth factor induces F9 cell proliferation and differentiation. AB - The 12S gene of the E1A region of adenovirus induces the production of a growth factor that is necessary for the immortalization of primary epithelial cells. This growth factor is able to replace the strict serum requirement of F9 cells, a teratocarcinoma-derived embryonal carcinoma cell line. Furthermore, the growth factor induces a morphological alteration in the F9 cells such that they resemble visceral endoderm cells and express the surface antigens, SSEA-1 and SSEA-3, characteristic of these differentiated cells. PMID- 2569708 TI - Amplification and rearrangement of c-erb B proto-oncogenes in cancer of human female genital tract. AB - There are two genes related to the v-erb B oncogene in the human genome. The c erb B-1 gene encodes the epidermal growth factor receptor (EGF-r), and the c-erb B-2/neu gene encodes a receptor-like protein very similar to, but distinct from the EGF-r. Southern blot analysis of DNAs from 15 fresh human ovarian carcinomas showed that the c-erb B-2/neu gene was amplified in 3 tumors. The c-erb B-1/EGF-r gene was not amplified. However, in one case of adenocarcinoma of uterine endometrium, the c-erb B-1/EGF-r was found to be rearranged in the 5' region of this gene. The genomic structure of the rearranged c-erb B-1/EGF-r gene is similar to those found in the chicken v-erb B oncogene, which produces a truncated form of the EGF-r. The results suggest that amplification of the c-erb B-2/neu gene may play a role in the pathogenesis of ovarian carcinoma. Further, the human c-erb B-1/EGF-r gene in adenocarcinoma of uterine endometrium may be activated by a similar mechanism as that in the chicken v-erb B oncogene. PMID- 2569709 TI - Characterization of buffalo-derived theilerial parasites with monoclonal antibodies and DNA probes. AB - The characteristics of intra-lymphocytic Theileria isolated from African buffalo and from cattle that were infected with buffalo-derived parasites were evaluated using anti-schizont monoclonal antibodies (mAbs) and DNA probes. Antigenic differences were revealed by the reactivities of 27 mAbs with the buffalo-derived parasites isolated from different animals. Antigenic diversity was also seen with Theileria-infected lymphoblastoid cell isolates taken from the lymph nodes and lambda gt11, showed specific hybridization to parasite DNA in Southern blots of restriction enzyme-digested, lymphoblastoid cells infected with buffalo-derived theilerial parasites. Genotypic differences between the buffalo-derived parasites were revealed by the restriction fragment length polymorphisms seen with hybridization of those probes to DNA from cloned and uncloned Theileria-infected cell lines. The evaluation of theilerial parasites derived from buffalo and from cattle which underwent typical T. p. lawrencei reactions, after being infected with buffalo-derived theilerial parasites, did not show any specific phenotypic or genotypic characteristics of these parasites that would distinguish them from T. p. parva and T. p. bovis parasites. The validity of these subspecies distinctions is discussed. PMID- 2569711 TI - Nucleotide sequence of the cDNA from the mouse leukocyte adhesion protein CD18. PMID- 2569710 TI - Sequence of the protease inhibitor domain of the A4 amyloid protein precursor of Mus domesticus. PMID- 2569712 TI - Ava I RFLP at the gas-1 locus on mouse chromosome 12. PMID- 2569713 TI - An EcoRI polymorphism in the human von Willebrand factor (vWF) gene. PMID- 2569714 TI - SacI and XbaI polymorphisms detected by lipocortin 2A (LPC2A). PMID- 2569715 TI - An arbitrary single copy DNA sequence VC85 [D1S85] detects a 500 bp insertion/deletion polymorphism on chromosome 1. PMID- 2569716 TI - Detection of a 3 allele AvaII RFLP by a single copy anonymous DNA sequence VC75 [D7S404] localized to chromosome 7. PMID- 2569717 TI - An arbitrary single copy sequence VC64 [D1S86] detects a moderate frequency TaqI RFLP on chromosome 1. PMID- 2569718 TI - An arbitrary single copy human DNA sequence VC63 [D4S129] detects a TaqI RFLP on chromosome 4. PMID- 2569719 TI - A high frequency two allele TaqI RFLP detected by an anonymous sequence VC61 [D2S65] on chromosome 2. PMID- 2569720 TI - High resolution deletion breakpoint mapping in the DMD gene by whole cosmid hybridization. AB - The locus DXS269 (P20) defines a deletion hotspot in the distal part of the Duchenne Muscular Dystrophy gene. We have cloned over 90 kilobase-pairs of genomic DNA from this region in overlapping cosmids. The use of whole cosmids as probes in a competitive DNA hybridization analysis proves a fast and convenient method for identifying rearrangements in this region. A rapid survey of P20 deletion patients is carried out to elucidate the nature of the propensity to deletions in this region. Using this technique, deletion breakpoints are pinpointed to individual restriction fragments in patient DNAs without the need for tedious isolation of single copy sequences. Simultaneously, the deletion data yield a consistent restriction map of the region and permit detection of several RFLPs. A 176 bp exon was identified within the cloned DNA, located 3' of an intron exceeding 150 Kb in length. Its deletion causes a frameshift in the dystrophin reading frame and produces the DMD phenotype. This exon is one of the most frequently deleted exons in BMD/DMD patients and its sequence is applied in a pilot study for diagnostic deletion screening using Polymerase Chain Reaction amplification. PMID- 2569722 TI - A new HincII RFLP for epidermal growth factor (EGF) on chromosome 4. PMID- 2569723 TI - Two TaqI dimorphic sites at the human beta-hydroxylase locus. PMID- 2569724 TI - RFLP for an anonymous single copy clone at 16pter-16p13.1 [D16S127]. PMID- 2569721 TI - Unique sequence organization and erythroid cell-specific nuclear factor-binding of mammalian theta 1 globin promoters. AB - The theta 1 globin gene is an alpha globin-like gene, and started to diverge from the other members of the alpha globin family 260 million years ago. DNA sequencing and transcriptional analysis indicated that it is functional in erythroid cells of the higher primates, but not in prosimians and rabbit. The theta 1 promoter region of higher primates including man consists of GC-rich sequences characteristic of housekeeping gene promoters, and CCAAT and TATA boxes located further upstream. It is shown here that the housekeeping gene promoter like region of human theta 1 contains two tandemly arranged, GC-rich motifs (GC-I and GC-II). Of these, GC-II interacts with nuclear factor(s) present in the globin-expressing, erythroleukemia cell line K562, before and after hemin induction. GC-I, however, interacts with nuclear factor(s) only present in hemin induced K562 cells. These factors are different from previously reported erythroid cell-specific factors, and are not detectable in non-erythroid Hela cells. Furthermore, the sequence of the motif GC-I and its location relative to ATG codon have been conserved among all known mammalian theta 1 globin genes. Finally, and most interestingly, the CCAAT box of theta 1 is contained within a 38 bp internal segment of Alu repeat sequence. Immediately upstream from this CCAAT box-containing Alu repeat segment is a 241 bp Alu repeat pointing in the opposite direction. The conservation of this novel arrangement among the higher primates suggests that an inserted Alu family repeat and its flanking genomic sequence have co-evolved, for at least 30 million years, to provide the canonical CCAAT and TATA promoter elements of the theta 1 globin genes in higher primates. PMID- 2569725 TI - DraI and PstI RFLPs in the tyrosine hydroxylase (TH) and insulin gene (INS) region of chromosome 11. PMID- 2569726 TI - A BanI polymorphism identified by the probe p alpha 1.4P at 19q13.1 [D19S37]. PMID- 2569727 TI - A Bsu36I RFLP detected by the chromosome 19q probe p alpha.5B [D19S36]. PMID- 2569728 TI - Isolation and mapping of a polymorphic DNA sequence (cA476) on chromosome 3 [D3S94]. PMID- 2569729 TI - TaqI and RsaI RFLP's at the prolactin inducible protein (PIP) locus on chromosome 7. PMID- 2569730 TI - [Efficacy of broxaterol (vs salbutamol) in asthma induced by physical exercise in children]. AB - B2-agonists play a first-rate role in the exercise induced asthma (EIA) prevention for their capacity to modulate the bronchial tone. We studied the efficacy of a new B-sympathomimetic (Broxaterol) in 10 children (7 males and 3 females) with EIA in double blind vs salbutamol. During two consecutive days a basal treadmill incremental exercise test was made for determining bronchoconstriction rate (IB). Subsequently the test was repeated 30' and 150' after the drug's administration. The Broxaterol premedication determined, by comparison with the basal IB value, a mean difference of 11.9 after 30' (p less than 0.05) and 10.0 after 150' (p less than 0.01). No significant correlation was found comparing the salbutamol premedication. In conclusion our data confirms the Broxaterol efficacy in exercise-induced asthma prevention. PMID- 2569731 TI - Solubility behavior of narcotic analgesics in aqueous media: solubilities and dissociation constants of morphine, fentanyl, and sufentanil. AB - The pH dependence of the aqueous solubility of morphine, fentanyl, and sufentanil was investigated at 35 degrees C. Dissociation constants and corresponding pKa' values of the drugs were obtained from measured free-base solubilities (determined at high pH's) and the concentrations of saturated solutions at intermediate pH's. Morphine, fentanyl, and sufentanil exhibited pKa' values of 8.08, 8.99, and 8.51, respectively. Over the pH range of 5 to 12.5 the apparent solubilities are determined by the intrinsic solubility of the free base plus the concentration of ionized drug necessary to satisfy the dissociation equilibrium at a given pH. Consequently, the drug concentrations of saturated aqueous solutions fall off precipitously as the pH is raised and ionization is suppressed. Further, at low pH's the aqueous solubility of morphine increased in a linear fashion with increases in the molar strength of citric acid which was added to acidify the medium, suggesting the formation of a soluble morphine citrate complex. PMID- 2569732 TI - Prenatal diagnosis of phenylketonuria by haplotype analysis. AB - Prenatal diagnosis of classic phenylketonuria (PKU) was performed after chorionic villus sampling by means of linked restriction fragment length polymorphisms (RFLPs) using the cDNA probe ph PAH 247 (Kwok et al. (1985) Biochemistry, 24, 556 561). We report in this paper a PKU family who were only informative for RFLP analysis by a combination of two RFLPs on the basis of haplotype determination of the normal and mutant phenylalanine hydroxylase (PAH) alleles. The DNA analysis detected a PKU fetus homozygous for mutant PAH alleles and the mother opted for termination in the 12th week of gestation. PMID- 2569733 TI - [Role of adrenoreceptors in the activation of the mouse hypothalamo-pituitary testicular complex induced by the presence of a female]. AB - A study was made of the role of the adrenergic mechanisms in the activation of endocrine testicular function of CBA/Lac and A/He male mice induced by the presence of a female in estrus without any tactile contact with a male. The alpha 1-adrenoreceptor blocker prazosin inhibited completely an increase in the peripheral blood plasma testosterone level caused by the receptive female challenge. At the same time, optimol blockade of beta 2-adrenoreceptors markedly increased the stimulating effect of the receptive female presence on the blood testosterone level. The expression of optimol action on the blood testosterone level depended on a male genotype. A conclusion was made that during sexual motivation in male mice caused by the presence of a receptive female, the stimulatory effects on testicular endocrine function were produced through the alpha 1-adrenoreceptors and the inhibitory effects were produced through the beta 2-adrenoreceptors. PMID- 2569734 TI - Characterization of variant gamma-glutamyl transpeptidase produced by pancreatocholangiocarcinoma cell lines in a protein-free, chemically defined medium. AB - Four pancreatocholangiocarcinoma cell lines (HPC-Y1, HPC-YT, MIA PaCa-2, and HChol-Y1) were established to propagate in a protein-free, chemically defined medium. High gamma-glutamyl transpeptidase (GGTP) activities were showed in their spent media (designated as the secreted (GGTP). Their GGTP activities in the spent media were 125, 85, 110, and 153 IU/L/mg of lyophilized spent media, whereas GGTP activities extracted from their cancer cell lines with bromelain were 105, 37, 86, and 112 IU/L/1 x 10(6) cells, respectively. The chemical characteristics of the GGTPs in the spent media from these cell lines resembled one of the GGTPs, sialic acid-rich GGTP, extracted from normal human pancreas with bromelain treatment as follows: the GGTPs secreted from the cancer cell lines bound to an anion exchange column moved fast on electrophoresis and then showed decreased electrophoretic mobility with neuraminidase treatment, showed a high affinity for concanavalin A and lentil lectin columns, and had an acidic isoelectric point. However, the elution patterns of erythroagglutinating phytohemagglutinin (E-PHA) column chromatography and thermostability tests demonstrated clear differences between the carcinoma GGTPs both in the spent media and cell lines and the sialic acid-rich GGTP of normal pancreas, namely the carcinoma GGTPs treated with neuraminidase showed affinity to E-PHA columns, and, in addition, the GGTPs in the spent media showed an apparent heat resistance at 56 degrees C. These findings indicate that the carcinoma GGTPs have a different oligosaccharide structure from that in normal pancreatic GGTPs. PMID- 2569735 TI - Pancreas transplantation: an immunohistochemical analysis of pancreatic hormones and HLA-DR expression. AB - The immunohistochemical features of pancreatic grafts in eight patients with pancreatic transplants were analyzed and compared with pancreases from five patients with chronic pancreatitis and with three pancreatic tissues without histological abnormalities. There was a significant increase in glucagon producing cells in patients with transplanted pancreases compared with those with chronic pancreatitis (P less than 0.05). A significant decline in insulin producing cells was seen in the transplanted pancreases with rejection in comparison with normal pancreatic tissue. Immunohistochemical staining for HLA-DR (Ia) antigens revealed expression of HLA-DR by endothelial cells, mononuclear cells, and by some ductal epithelial cells, but not by the endocrine islet cells. These results suggest that significant changes in insulin and glucagon production occur in the transplanted pancreas with rejection and that HLA-DR is not expressed by islet cells during graft rejection or with chronic inflammation. PMID- 2569736 TI - Morphologic differences between polyarteritis and Wegener's granulomatosis using light, electron and immunohistochemical techniques. AB - The differential diagnosis of renal biopsies of patients with polyarteritis nodosa (PAN) versus those with Wegener's granulomatosis (WG) is difficult because in both the morphologic expression is a focal proliferating and necrotizing glomerulonephritis (GN) with crescents. Twenty-nine biopsies of PAN and 29 of WG were studied by light, electron microscopy and immunohistochemistry. Whereas in PAN the dominent feature is severe focal fibrinoid necrosis of glomerular tufts, WG is characterized by destructive extracapillary GN with capsular breaks and periglomerulitis leading to complete glomerular destruction and replacement by a localized inflammatory exudate in the form of a granuloma. In PAN necrotizing arteritis is common (19 of 29), whereas only 1 of 29 cases of WG showed arteritis. Depending on the stage of the disease, various types of hematogenous cells can be seen in both. In general, the dominent cell type in PAN is polymorphonuclear cells (PMNs) with or without eosinophils, whereas in WG mononuclear cells dominate the picture. Monocytes and macrophages, including giant cells in the crescents, periglomerular infiltrates, and in the granulomas are seen, whereas in the interstitium, plasma cells predominate. The histologic ultrastructural and immunohistochemical findings in both groups will be discussed. PMID- 2569737 TI - Association of charge clusters with functional domains of cellular transcription factors. AB - Using rigorous statistical methods, we have identified and evaluated unusual properties of the distribution of charged residues within the sequences of eukaryotic cellular transcription factors. Virtually all transcription factors, including GAL4, c-Jun, C/EBP, CREB, Oct-1, Oct-2, Sp1, Egr-1, CTF-1, steroid and thyroid hormone receptors, and others, carry one or more highly significant charge clusters. For the most part these clusters (conserved within families of homologous proteins) are of positive net charge but contain also substantial numbers of acidic residues. Predominantly basic charge clusters are often, but not exclusively, associated with DNA-binding domains, and vice versa. Negative charge clusters of note occur only in the yeast protein PHO4 and in the proteins encoded at the Drosophila loci zeste (zeta) and knrl. This dearth of statistically significant negative charge clusters raises questions with respect to the generality of acidic activation domains. A number of sequences (Oct-1, Oct 2, zeste, Dhr23, E75, and knrl) contain multiple charge clusters together with one or more significantly long uncharged regions. The occurrence of multiple charge clusters is a rare phenomenon (found in less than 3% of all proteins, mainly in Drosophila developmental control proteins and in transactivators of eukaryotic DNA viruses). Most of the proteins with zinc-binding "fingers" carry a mixed charge cluster centered at the zinc-finger motif preceded by a long uncharged stretch, suggestive of a modular structure for these proteins. PMID- 2569738 TI - Yeast TATA-binding protein TFIID binds to TATA elements with both consensus and nonconsensus DNA sequences. AB - The DNA binding properties of the yeast TATA element-binding protein TFIID were investigated. The affinity (apparent equilibrium dissociation constant) of TFIID for the adenovirus major late promoter consensus TATA element is 2 x 10(-9) M, a value similar to the affinity of gene-specific regulatory proteins for their binding sites. TFIID binding is highly specific and recognizes nonspecific sites with approximately 10(5)-fold lower affinity. Despite this specificity, TFIID also binds with high affinity to several TATA elements that do not match the consensus TATA sequences (TATAAA and TATATA): the yeast LEU2 TATA (TATTATTTA), the simian virus 40 TATA (CTTATTTAT), and the yeast CYC1 -10 TATA (TTATACATT) all bound TFIID. Furthermore, TFIID was active in promoting transcription in vitro from the nonconsensus TATA elements. Thus, contrary to previous suggestions, the existence of nonconsensus TATA elements does not itself indicate the existence of multiple TATA-binding factors. PMID- 2569739 TI - Fatal syndrome in mice engrafted with cells producing high levels of the leukemia inhibitory factor. AB - Cells of the murine hemopoietic cell line FDC-P1 were multiply infected with a retroviral construct containing cDNA encoding the leukemia inhibitory factor (LIF) to produce cells secreting high levels of LIF. Injection of these cells to unirradiated or irradiated syngeneic DBA/2 mice resulted in animals engrafted with LIF-producing cells in the marrow, spleen, and lymph nodes and with elevated serum LIF levels. These mice developed within 12-70 days a fatal syndrome characterized by cachexia, excess new bone formation, calcification in heart and skeletal muscle, pancreatitis, thymus atrophy, and abnormalities in the adrenal cortex and ovarian corpora lutea. Injection of mice with control FDC-P1 cells led to comparable organ engraftment, but the mice developed none of these lesions. The observations suggest that LIF may be a potent cachexia-inducing agent and may have marked effects on osteoblasts and calcium metabolism. PMID- 2569740 TI - Absence of unsaturated fatty acid synthesis in murine T lymphocytes. AB - Stearic acid is toxic for T lymphocytes in vitro but has little effect on B lymphocytes. To investigate the molecular basis for this difference, purified murine T and B lymphocytes were compared for their abilities to incorporate and metabolize stearic acid. Unstimulated T and B cells incorporated identical amounts of stearic acid into six different phospholipids and four neutral lipids. After mitogen stimulation, fatty acid uptake was increased in both lymphocyte types, but cell-specific differences were seen in the distribution of stearic acid among the various cellular lipids. Doses of stearic acid that selectively inhibited T-cell proliferation resulted in a 5-fold greater accumulation of distearoylphosphatidylcholine in T cells than in B cells. Whereas T cells did not desaturate the exogenously derived stearic acid, up to 25% of the saturated fatty acid was converted to oleic acid in B cells. These findings suggested a deficiency of stearoyl-CoA desaturase (acyl-CoA, hydrogen-donor:oxygen oxidoreductase, EC 1.14.99.5) activity in T cells, which was confirmed by subsequent studies. Cell-free extracts from B cells displayed nearly 20-fold more stearoyl-CoA desaturase activity than T-cell extracts, and the level of stearoyl CoA desaturase mRNA was 30-fold higher in B cells. Collectively, our data indicate that murine T cells are deficient in unsaturated fatty acid synthesis. The deficiency of stearoyl-CoA desaturase in T cells may represent the basis for the differing sensitivities of T and B lymphocytes to inhibition by saturated fatty acids. PMID- 2569742 TI - Linkage analyses in families with nephrogenic diabetes insipidus. PMID- 2569743 TI - Characterization of new probes for diagnosis of polycystic kidney disease (PKD1). PMID- 2569741 TI - A family of cyclin homologs that control the G1 phase in yeast. AB - Two Saccharomyces cerevisiae genes were isolated based upon their dosage dependent rescue of a temperature-sensitive mutation of the gene CDC28, which encodes a protein kinase involved in control of cell division. CLN1 and CLN2 encode closely related proteins that also share homology with cyclins. Cyclins, characterized by a dramatic periodicity of abundance through the cell cycle, are thought to be involved in mitotic induction in animal cells. A dominant mutation in the CLN2 gene, CLN2-1, advances the G1- to S-phase transition in cycling cells and impairs the ability of cells to arrest in G1 phase in response to external signals, suggesting that the encoded protein is involved in G1 control of the cell cycle in Saccharomyces. PMID- 2569745 TI - [Workgroup of European Nurse Researchers]. PMID- 2569744 TI - DNA marker analysis of adult polycystic kidney disease in Italian families. Italian Cooperative Group on ADPKD. PMID- 2569746 TI - Neurochemical and receptor theories of depression. AB - The neurochemical and receptor theories relate depression to deficient neurotransmission at critical sites in the brain. While this concept has generated a number of theories of depression over the years, the research findings do not fully support any single theory in its entirety. Several issues thus remain controversial or inconclusive. For instance, the monoamine deficit theory is supported by low urinary MHPG in some forms of bipolar, but not unipolar depression. Cerebrospinal fluid MHPG and 5-HIAA studies are inconclusive. Amine metabolite research is also limited in scope because the information derived pertains to pre-synaptic and synaptic events and ignores post synaptic events. Receptor research, which includes study of both pre-and post synaptic sites, suggests supersensitivity of Beta-adrenergic receptors in depression. But this research is criticized because it is mostly animal based. Also, the findings of low melatonin in depression contradict the supersensitivity hypothesis. Abnormally low post-synaptic alpha-2 adrenoceptors is indicated by findings of an attenuated GH response to clonidine. But abnormality of pre synaptic alpha-2 adrenoceptor functions has not been demonstrated conclusively. Recent findings in depression suggest a dysregulation in the dynamic and interactive relationship between neurotransmitters and receptors. Accordingly, a comprehensive view of the abnormalities of the various neurotransmitter systems in depression requires studies which investigate pre- and post-synaptic events simultaneously, preferably during illness and remission. PMID- 2569747 TI - The neurochemistry of mood. PMID- 2569748 TI - The acute treatment of anxiety and depression. AB - Anxiety and depression are commonly occurring symptoms. Anxiety disorders and mood disorders usually share common symptoms and they frequently co-exist. There is a considerable body of research that has demonstrated that anxiety and depression can be distinguished from each other at the syndrome level. There is also evidence that such a distinction is arbitrary and not well substantiated. Clinically, the practitioner is often faced with the problem of treating a patient who presents with anxiety and depressive symptoms at the same time. It is well-established that the first line of treatment in major mood disorder is the used of tricyclic antidepressant in adequate dosage. The first line of treatment for the anxiety disorders is usually the administration of benzodiazepine anxiolytics. The anti-depressants have to be given for some months to the majority of patients whereas the anxiolytics are given for short periods. The tricyclics have a relatively slow onset of action compared to the benzodiazepines. Recent evidence is available about the effectiveness of the triazolo-benzodiazepines in panic disorder with or without secondary major mood disorder. There are also reports of the effects of the triazolo-benzodiazepines in primary mood disorder. In these mood disorders, the benzodiazepines caused rapid relief of both anxious and depressive symptomatology. The effects of the benzodiazepines occur even in the presence of melancholic depression. Where anxiety and depression coexist, the clinician may wish to consider beginning anti depressant therapy with combined tricyclic antidepressant and benzodiazepine to produce rapid symptom relief. After four weeks the benzodiazepine should be faded out and therapy continued with the tricyclic medication alone. PMID- 2569749 TI - On the neurobiology of depression: research based on heterogeneous diagnostic groups produces heterogeneous biological data. AB - An improvement in the accuracy and specificity of the criteria for identifying people with major depressive disorder would be of great benefit not only in the diagnosis and treatment of patients with depression, but also in research concerning the biological substrates of the emotions. Unfortunately, attempts at developing a biological diagnostic test for depression based on the analysis of major depressive disorder patients identified on the basis of existing diagnostic criteria have not been successful. Undoubtedly, this is due in part to the complexity of the neurochemistry and neuroanatomy of the emotions. But more importantly, it is due to the broad global nature of the criteria used to identify depressed patients. This results in the biological study of patients who are similar in general terms but who differ in specific behavioral symptoms and underlying neurobiology. A detailed analysis of the behavior of depressed people might reveal subtle differences that could be used to separate patients with depression into more homogeneous subgroups for biological study. This would increase the probability of developing biological tests that would lead to further refinement in the diagnosis of depression and in the selection of the most appropriate therapeutic intervention for a particular patient. PMID- 2569750 TI - Limited eye movement patterns in chronic schizophrenic patients. AB - Eye movements of 29 chronic schizophrenic patients and 23 normal control subjects were measured by an eye mark recorder during viewing of stationary geometric figures. The results suggested that the eye movements of schizophrenic patients were more limited than those of normal control subjects and that the abnormal eye movement inspection patterns in schizophrenic patients may reflect a lack of visual investigation of novel stimuli in the environment. Our findings, together with Moriya's (1979) observation of similar eye movement pattern abnormalities in the relatives of schizophrenic patients, support the hypothesis that the limited eye movement patterns may be a biological marker for schizophrenia. PMID- 2569751 TI - Neuroleptic nonresponse and affective symptoms: a 2-year prospective study of schizophrenic outpatients. AB - For 2 years we assessed clinical state, and plasma neuroleptic and prolactin levels every 6 months in 105 male schizophrenic outpatients. The patients took a variety of neuroleptics at clinically determined doses. Those who had psychotic symptoms at 50% or more of their visits had higher neuroleptic doses, more tardive dyskinesia, and more affective symptoms than the other patients. These groups were not, however, significantly different in their age of onset, years of education, duration of illness, duration of neuroleptic exposure, or negative symptoms. PMID- 2569752 TI - Onset and course of tardive dyskinesia. PMID- 2569753 TI - [Renal artery aneurysm in Wegener's granulomatosis]. AB - Microaneurysms of renal and visceral arteries are characteristic signs of periarteritis nodosa. Normally they are not found in Wegener's disease, where glomerulonephritis is commonly observed. We report on a patient with vasculitis of the upper and lower respiratory tract, focal glomerulonephritis, prostatic and pulmonary granulomas and anti-cytoplasm antibodies corresponding to Wegener's disease. The most striking findings in angiography were multiple small aneurysms of the peripheral branches of the renal artery. At necroscopy these angiographic findings were histologically proven as necrosis of the arterial wall with destruction of the elastic lamina, causing local vascular ectasia. These renal vascular changes are a characteristic sign of periarteritis nodosa. The combination of clinical, laboratory, radiologic and histologic findings in our patient can be explained as an overlap-syndrome of Wegener's disease and periarteritis nodosa. We assume that the combination of pathologic findings in our patient correspond to a rare atypical renal manifestation of Wegener's disease. PMID- 2569754 TI - [A case of prolonged effect of dipotassium chlorazepate and its reversal by flumacenil]. PMID- 2569755 TI - The effect of UK14,304-18 (an alpha-2 adrenergic agonist) on myocardial blood flow during cardiopulmonary resuscitation. AB - Several recent studies have suggested that adrenergic drugs with peripheral postsynaptic alpha-2 agonist properties increase aortic diastolic pressure (ADP), and thus in the setting of CPR, may improve myocardial blood flow (MBF). This preliminary study investigated the effect of UK14,304-18, a postsynaptic alpha-2 adrenergic agonist on ADP, MBF, myocardial oxygen delivery/utilization (MDO2/MVO2), endocardial/epicardial blood flow ratio (EN/EP), coronary sinus oxygen content (CcsO2) and extraction ratio (ER) during CPR. Five swine were instrumented for MBF measurements using tracer microspheres. Catheters were also placed to measure arterial oxygen content (CaO2) and CcsO2. ADP, MBF, MDO2/MVO2, EN/EP, ER, CaO2 and CcsO2 were measured during normal sinus rhythm (NSR), and during CPR following a 10-min cardiorespiratory arrest. Following this, each animal received 2.0 mg/kg of UK14,304-18 through a right atrial line. ADP, MBF, MDO2/MVO2, EN/EP, ER, CaO2 and CcsO2 were again determined. Defibrillation was then attempted. To determine whether UK14,304-18 improved ADP, MBF and MDO2 over MVO2, compared to CPR alone, results were compared using a paired Student t-test. Statistical significance was considered at the P less than or equal to 0.05 level. No significant improvement in ADP, MBF, MDO2 or ER was noted following the administration of UK14,304-18. The lack of improvement in ADP and MBF may be secondary to a centrally acting postsynaptic alpha-2 agonist effect because of disruption of the blood brain barrier following a prolonged cardiac arrest or because of pharmacologically or structurally distinct populations of peripheral postsynaptic alpha-2 adrenoreceptors that develop in this setting.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569756 TI - [Mixed cryoglobulinemia with necrotizing angiitis. Apropos of 2 cases]. AB - Out of a series of 26 personal cases, 2 cases of mixed IgM-IgG cryoglobulinemia, one type II the other type III, are reported because they were associated with histologically proven necrotizing vasculitis. In both cases the numerous symptoms were due to renal damage (the vasculitis was discovered in the kidney) and to peripheral neuropathy. One of the patients died; the other had severely deteriorated general condition and required substitution hemodialysis. Cases of vasculitis associated with mixed cryoglobulinemia have often been published, but there are few reports mentioning necrotizing vasculitis; a search in the literature yielded only 9 cases. This small number does not mean that mixed cryoglobulinemia should not be listed among the causes of necrotizing vasculitis, but it makes it difficult to extract those specific features that would enable to predict which case of mixed cryoglobulinemia is associated or not with necrotizing vasculitis. PMID- 2569757 TI - [Renal involvement in necrotizing angiitis]. PMID- 2569758 TI - [Gamma GT assay: a necessary and sufficient test for the screening of hepatopathies?]. PMID- 2569759 TI - Diagnosis of hemophilia A in a female subject by using restriction fragment length polymorphisms linked to the factor VIII gene. AB - A 6-year-old girl, daughter of a male patient with moderate hemophilia A (factor VIII 3%), was referred to our Center because she also had very low levels of factor VIII (4%). The proband's brother has mild hemophilia A (7%); the mother (29%) is a possible carrier, no other case of hemophilia A being reported in her family. Hence, the factor VIII deficiency found in the girl is consistent either with a carrier state with extreme lyonization in favour of the hemophilic gene or with homozygosity for the hemophilia gene. To distinguish these possibilities, we studied the segregation of three restriction fragment length polymorphisms (RFLPs) linked to the factor VIII gene in the 4 members of the family. Employing one intragenic (FVIII-BclI) and two extragenic (St14-TaqI and Dx13-BglII) RFLPs, we showed that the proband has inherited from both parents the defective gene, being therefore homozygous for the hemophilia gene. PMID- 2569760 TI - NcoI restriction fragment length polymorphism (RFLP) of the tumour necrosis factor (TNF alpha) region in primary biliary cirrhosis and in healthy Danes. AB - The restriction fragment length polymorphism of the human tumour necrosis factor (TNF alpha) region was investigated by means of 20 different restriction enzymes and a human TNF alpha cDNA probe. Only one of the enzymes, NcoI, revealed a polymorphic pattern consisting of fragments of 10.5 and 5.5 kb, which behaved as alleles. In a panel of 108 random, healthy, unrelated Danes, the phenotype frequencies of the 10.5 and 5.5 kb bands were 0.94 and 0.53 and the gene frequencies were 0.71 and 0.29, respectively. The test for Hardy-Weinberg equilibrium showed no significant deviation from the expected values. The 5.5 kb band was strongly positively associated with HLA-DR3, HLA-B8, and HLA-A1. In 22 patients with primary biliary cirrhosis a significantly (corrected P = 0.024) decreased frequency of the 10.5 kb fragment was found. Additional studies are in progress to substantiate this association. PMID- 2569761 TI - Variation in cadmium content of caribou tissues from northern Quebec. AB - Samples of kidney, liver, skeletal muscle, heart, mesentery and rumen wall were collected during 1985-1986 in northern Quebec from 121 caribou belonging to the Riviere George herd. Significant seasonal variations were found for cadmium concentration in kidneys, liver and skeletal muscles, the level being higher in winter than in autumn. The concentration in liver and kidneys increased with age, while the effect of this factor was complex in skeletal muscles. Sex-related differences were found in muscle and liver levels of cadmium. According to season, sex and age, mean dry-weight concentrations varied between 5.6 and 51.3 micrograms g-1 in kidneys, 1.7 and 4.0 micrograms g-1 in liver and 0 and 0.27 microgram g-1 in skeletal muscles. Season and sex-related differences could be explained by seasonal variation in cadmium intake and by the annual weight cycle exhibited by caribou kidneys and liver. Because of the relatively high cadmium content, it was recommended not to consume liver and kidneys of free-ranging caribou in Quebec; this restriction did not apply to meat, heart, mesentery or rumen wall that contained limited concentrations of this heavy metal. PMID- 2569762 TI - NMDA antagonists differentiate epileptogenesis from seizure expression in an in vitro model. AB - In an electrographic model of seizures in the hippocampal slice, both of the N methyl-D-aspartate (NMDA) antagonists 2-amino-5-phosphonovaleric acid and 5 methyl-10,11-dihydro-5H-dibenzo(a,d)cyclohepten-5,10-imine maleate (MK-801) prevented the progressive development of seizures but did not block previously induced seizures. Thus, a process dependent on the NMDA receptor-ionophore complex establishes a long-lasting, seizure-prone state; thereafter the seizures depend on non-NMDA receptor-ionophore mechanisms. This suggests that there is an important distinction between epileptogenesis and seizure expression and between antiepileptogenic and anticonvulsant pharmacological agents. PMID- 2569763 TI - A novel mRNA of the A4 amyloid precursor gene coding for a possibly secreted protein. AB - The gene, encoding the A4 peptide found in the amyloid core of senile plaques isolated from the cerebral cortex of patients with Alzheimer's disease, produces at least three precursors that resemble cell surface receptors. A clone isolated from a human brain complementary DNA library contained the structural sequence for an A4 amyloid peptide precursor with a serine protease inhibitor domain in which 208 amino acids at the carboxyl terminal are replaced by 20 amino acids derived from nucleotide sequences with homology to the Alu repeat family. This protein devoid of the transmembrane domain most likely represents a secreted form of the A4 amyloid peptide precursor. PMID- 2569764 TI - [Effects of noradrenaline,glycine, gamma-aminobutyric acid and L-glutamic acid on spinal field potentials in the rat]. AB - Noradrenaline (NE), glucine (GLY), gamma-aminobutyric acid (GABA) and L-glutamic acid (L-GLU) were locally applied to the dorsal surface of lumbar spinal cord on anesthetized and immobilized rats and their effects on segmental (SP) and descending (DP) spinal field potentials were examined. Both waves N and P of SP and DP were reduced markedly in amplitudes following local application of NE. A significant decrease in amplitude of wave N and a remarkable increase in wave P in both SP and DP were shown during the application of GLY, GABA and L-GLU. The results suggest that these neurotransmitters may be involved in the generation of waves N and P, particularly affecting the activities of interneurons in the spinal dorsal horn. PMID- 2569765 TI - Human gene for proliferating cell nuclear antigen has pseudogenes and localizes to chromosome 20. AB - We have isolated from a human genomic library a pseudogene of the proliferating cell nuclear antigen (PCNA) gene. Its sequence shows a 78% similarity with the human PCNA/cDNA. The PCNA gene is located on human chromosome 20, while the pseudogene maps to chromosome region Xpter in equilibrium Xq13. An additional locus detected by the full-length PCNA cDNA, but not by intron probes, segregates concordantly with chromosome region 6p12 in equilibrium 6pter and probably represents a second pseudogene. PMID- 2569766 TI - Structure of four amplified DNA novel joints. AB - The structures of four novel joints present in the amplified DNA of a Syrian hamster cell line highly resistant to N-(phosphonacetyl)-L-aspartate were analyzed. Novel joints J1, J2, and J4 were formed by recombination between two regions of wild-type DNA, whereas joint J3 is the end point of an inverted duplication. A fraction of the J3 copies displays a cruciform structure in the purified genomic DNA. The formation of J1 and J2 apparently involved a simple breakage and joining of the two wild-type sequences, whereas extra nucleotides are present at the junction point of J3 and J4. The two regions of the wild-type DNA which have recombined to form J1, J2, and J4 show few sequence similarities, indicating that these joints probably resulted from nonhomologous recombination. AT-rich regions are present in the vicinity of the breakpoint for the four joints and eight of 10 crossover points could be associated with putative topoisomerase I cleavage sites. Our results indicate that different types of novel joints are present in the amplified DNA of this cell line, which was isolated after several steps of selection. PMID- 2569767 TI - Genes for beta 2-adrenergic receptor and platelet-derived growth factor receptor map to mouse chromosome 18. AB - The beta 2-adrenergic receptor (ADRB2R) mediates the response of various cel types to neurotransmitters, hormones, and drugs. The platelet-derived growth factor (PDGF) interacts with its receptor (PDGFR) to stimulate mesenchymal cell proliferation. In the human, ADRB2R and PDGFR have been mapped to the q31--q32 region of chromosome 5 (HSA5). Here we report the mapping of Pdgfr and Adrb2r to mouse chromosome 18 (MMU18) using somatic cell hybrid mapping techniques. Together with previous mapping of genes for the glucocorticoid receptor (human locus GRL; mouse locus Gr1-1), the class II HLA invariant chain (human locus PHLAG; mouse locus Ii) and the FMS protooncogene to HSA5 and MMU18, the assignment of both Pdgfr and Adrb2r to MMU18 expands the conserved autosomal syntenic group. PMID- 2569768 TI - [Non-hormonal treatment in the menopause]. PMID- 2569769 TI - Inhibition of growth of human breast carcinomas in vivo by somatostatin analog SMS 201-995: treatment of nude mouse xenografts. AB - Minced tumor fragments were xenografted into subcutaneous tissue of the lateral thoracic regions of young adult, virgin female nude mice to study the effects of somatostatin analog SMS 201-995 on growth of estrogen-dependent (MCF-7) and estrogen-independent (BT-20) human breast carcinomas. When tumors became palpable (6 to 10 days), mice were assigned randomly to receive either SMS (4 to 50 micrograms) or acetate buffer (0.2 ml) subcutaneously twice a day. For MCF-7, mean tumor volume was significantly lower on day 20 and days 30 through 50 in SMS treated mice than in controls (p less than 0.05), and tumor doubling time was increased from 13.2 to 19.0 days. Calculated growth increment was significantly lower with SMS than with buffer treatment (1.1 +/- 0.1 vs 1.9 +/- 0.2) (p less than 0.001). For BT-20, mean tumor volume of SMS-treated mice was slightly, but not significantly, lower than that of controls; however, calculated growth increment was significantly lower for SMS treatment (3.2 +/- 0.3 vs 3.9 +/- 0.4) (p +/- 0.001), and tumor doubling time was increased from 4.0 to 5.8 days. For MCF-7, flow cytometric DNA analysis of tumor biopsy samples demonstrated a reduced G2 + M phase with SMS treatment. We conclude that SMS slows the growth of both MCF-7 and BT-20 human breast cancer xenografts in nude mice and that SMS may be clinically useful in the management of patients with breast carcinoma. PMID- 2569770 TI - [International cooperation. ICN Congress: nursing's "United Nations"]. PMID- 2569771 TI - [International cooperation. Resolutions from the ICN Congress]. PMID- 2569772 TI - [Psychiatry. Other and more than neuroleptics, thanks!]. PMID- 2569773 TI - [Biochemical tests in the diagnosis of nephrotoxicity: assessment of the significance]. AB - In order to identify early signs of nephrotoxicity in 38 patients with malignant tumors of the testicle, the data of 3 biochemical tests (measurements of gamma glutamyltranspeptidase (gamma-GT) activity in the urine, of the content of creatinine and urea in blood serum) were compared. Activation of gamma-GT in the urine may be viewed as the first and early sign of nephrotoxicity, since the rise of creatinine and urea in the blood was observed later. The magnitude of gamma-GT activity (means + 2 sigma) - 5.9 Units/mmo le Cr may provide basis for the judgement about nephrotoxicity according to the scheme applied. The fall of gamma GT activity without any further increase after multiple courses of chemotherapy is of prognostic significance for its evidences profound injury to the renal parenchyma and may serve as basis for correcting the doses of chemotherapeutic drugs. The reduction of gamma-GT activity to less than 3 Units/mmole creatinine (means - 2 sigma) may be regarded as indication for instituting disintoxication therapy. PMID- 2569774 TI - [Circadian rhythms of mineral-excreting kidney function in healthy persons and their changes in ischemic heart disease]. AB - Seventy normal persons and 100 patients with coronary disease were examined for circadian rhythms of urine and minerals excretion. The data obtained demonstrate that normal persons were characterized by circadian rhythms of urine and minerals excretion with a definite confidence interval of mesor and amplitude fluctuations. Angina pectoris of effort was marked by infradian rhythms of mineral excreting renal function. Monotherapy with beta-blockers or calcium antagonists given in courses entailed the recovery of the circadian rhythms of mineral-excreting renal function. Parameters of the rhythms of urine and minerals excretion in patients with angina pectoris of effort and rest noticeably differed from those in patients with angina pectoris of effort. In 30.7 percent of the persons, the use of the mathematic methods did not reveal any significant rhythms. The circadian range was found to predominate among the significant rhythms. Antianginal treatment did not produce any appreciable changes. PMID- 2569775 TI - Transglutaminase-catalysed cross-linking: a potential mechanism for the interaction of fibrinogen, low density lipoprotein and arterial type III procollagen. AB - Bovine type III [3H]procollagen or its [125I]aminopropeptide were shown by chromatography under dissociating conditions to form very high molecular weight compounds with excess bovine fibrinogen after incubation with purified tissue transglutaminase, though none is formed with other major plasma proteins. Larger compounds of this type formed from fibrinogen or fibrin monomer can be separated by centrifugation and they are insoluble on washing with 1% SDS. Ultracentrifugation showed that a significant fraction of [3H]procollagen III forms a low density complex on incubation with transglutaminase plus excess IDL or LDL, but not HDL. SDS polyacrylamide gel electrophoresis showed that type III collagen [125I]aminopropeptide forms high molecular weight compounds after incubation with transglutaminase plus excess IDL or LDL but not with HDL. It is hypothesized that, in the presence of excessive concentrations of LDL and/or fibrinogen and of tissue transglutaminase, crosslinking reactions of the type demonstrated may interfere with normal injury-repair processes and stimulate the formation of atherosclerotic lesions in arteries. PMID- 2569776 TI - Peptide inhibition of accelerated transplant atherosclerosis. PMID- 2569777 TI - Peptide inhibition of myointimal proliferation following angioplasty in rabbits. PMID- 2569778 TI - Inhibition of 3H-thymidine incorporation by angiopeptin in the aorta of rabbits after balloon angioplasty. PMID- 2569779 TI - Angiopeptin inhibits thymidine incorporation in rat carotid artery in vitro. PMID- 2569780 TI - Hpa-I polymorphism and the sickle gene in Nigerians. AB - The Hpa-I restriction fragment length polymorphism linked to the beta globin genes was studied in 181 Nigerian subjects. The beta S gene was found to be linked to the 13 kilobase (kb) Hpa-I fragment in 98.4 percent while 1.6 percent was linked to the Hpa-I 7.6 kb fragment. The majority of beta A genes (94.2%) was found to be linked to either the 7.6 or 7.0 kb fragment whilst the remainder (5.8%) was linked to the 13.0 kb. The beta A 13 kb linkage and the small proportion of the beta S genes linked to the 7.6 kb fragment would lessen the accuracy of the Hpa-I polymorphism in the prenatal diagnosis of sickle cell anaemia in this population. PMID- 2569781 TI - Subcellular distribution of titanium in the liver after treatment with the antitumor agent titanocene dichloride. A study using electron spectroscopic imaging. AB - In the present study, the subcellular distribution of titanium in the liver of mice was determined 24 and 48 h after application of a therapeutic (ED100; ED = effective dose) and a toxic (LD25; LD = lethal dose) dose (60 and 80 mg/kg, respectively) of the antitumor agent titanocene dichloride by electron spectroscopic imaging at the ultrastructural level. At 24 h, titanium was mainly accumulated in the cytoplasm of endothelial and Kupffer cells, lining the hepatic sinusoids. Titanium was detected in the nucleoli and the euchromatin of liver cells, packaged as granules together with phosphorus and oxygen. One day later titanium was still present in cytoplasmic inclusions within endothelial and Kupffer cells, whereas in hepatocyte nucleoli only a few deposits of titanium were observed at 48 h. At this time titanium was mainly accumulated in the form of highly condensed granules in the euchromatin and the perinucleolar heterochromatin. It was found in the cytoplasm of liver cells, incorporated into cytoplasmic inclusion bodies which probably represent lysosomes. Sometimes these inclusions were situated near bile canaliculi and occasionally extruded their content into the lumen of bile capillaries. This observation suggests a mainly biliary elimination of titanium-containing metabolites. These results confirm electron spectroscopic imaging to be an appropriate method for determining the subcellular distribution of light and medium-weight elements within biological tissues. Insights into the cellular mode of action of titanocene complexes or titanocene metabolites can be deduced from the findings of the present study. PMID- 2569782 TI - Correlations between blood glucose levels and bromodeoxyuridine labelling indices of pancreatic islet cells following streptozotocin administration to pregnant Syrian golden hamsters. AB - Eighteen timed-pregnant Syrian golden hamsters were injected subcutaneously with streptozotocin (STZ, 60 mg/kg bw) early on gestational day 10. The response varied widely, and based on changes in blood glucose levels during gestational days 11 to 15, the hamsters were categorized into four groups: 1) no change; 2) mild diabetes (200-250 mg/dl), which reverted; 3) moderate diabetes (greater than 300 mg/dl), which reverted; and 4) moderate to severe diabetes (300-500 mg/dl), which was sustained. Two hours before sacrifice, a 25 mg tablet of bromodeoxyuridine (BrdU) was implanted subcutaneously into each experimental hamster and into 17 control pregnant hamsters that had not received STZ. BrdU labelling was demonstrated immunochemically in the pancreatic islet cells. In control hamsters, the mean labelling index (LI) of the islet cells was 0.07% and did not exceed 0.2% in any hamster. Following injection of STZ, islet cell LI's remained low (0.13%) if the blood glucose levels were not altered by the diabetogenic drug. However, LI's were increased in islet cells of hamsters which showed a mild to moderate diabetes which rapidly reverted; the highest LI's (5% +/- 2.1) occurred in four hamsters that were killed 2 days after receiving STZ. The LI's were moderately increased (1.4% +/- 0.42) in two hamsters with moderate diabetes killed 2 days after STZ, but LI's were low (0.12% +/- 0.04) in six hamsters with moderate to severe diabetes killed 3, 4, and 5 days after STZ. Reversion of hyperglycemia to normoglycemia correlated closely with increased DNA synthesis in the islet cells of the pregnant hamsters. These observations strongly suggest that following mild cytotoxic injury induced by STZ, the B cells regenerated and insulin production was restored sufficiently to maintain normoglycemia. PMID- 2569783 TI - The effect of bilateral amygdaloid nucleus lesions on rat small bowel crypt cell kinetics. AB - Previous investigators have found that central nervous system lesions, in particular lesions of the hypothalamus, may increase the crypt cell mitotic rate in the rat small bowel. Since the amygdaloid nuclei form part of the limbic system (the "visceral brain") and have functional neural connections with the hypothalamus the effect of bilateral electrocoagulation lesions of the amygdaloid nuclei on crypt cell mitotic rate in the rat small bowel was investigated, using a stathmokinetic technique. Bilateral amygdaloid lesions were found to be associated with a marked increase in crypt cell mitotic rate in the proximal jejunum and distal ileum. Consideration of the neural connections of the amygdaloid nuclei suggests that these effects may possibly be mediated via the hypothalamus and the autonomic nervous system. The effects of lesions of other parts of the limbic system on crypt cell mitotic rate will be published subsequently. PMID- 2569784 TI - Macrophages of the rat thymus after cyclosporin treatment. Histochemical, enzymehistochemical and immunohistochemical study. AB - Young adult Wistar rats received 40 mg/kg of cyclosporin perorally for 21 days. Cyclosporin induced almost total disappearance of thymic medulla, whereas the cortex remained preserved. Although the density of cortical macrophages did not change significantly, their characteristics altered markedly and they became enlarged and rounded. In addition to an increase in acid phosphatase and nonspecific esterase activities, cortical macrophages developed very strong succinic dehydrogenase and chloroacetate esterase activities and a fine, granular, aldehyde fuchsin-positive cytoplasmic content. However, these cytoplasmic granules were PAS-negative and were not sudanophilic. Cortical macrophages retained their normal antigenic properties (which were studied by the use of ED1, ED2 and R-MC 41 monoclonal antibodies). Phagocytic cells in the remaining medullary islands retained their usual characteristics. The changes in cortical macrophages after cyclosporin treatment are discussed, especially in relation to the characteristics of macrophages of the cortico-medullary zone in the normal rat thymus. PMID- 2569785 TI - Biochemical microanalysis of alpha-glucosidase activity in preneoplastic and neoplastic hepatic lesions induced in rats by N-nitrosomorpholine. AB - Preneoplastic and neoplastic hepatic lesions were induced in male Sprague-Dawley rats by oral administration of N-nitrosomorpholine (NNM) for 7 weeks at a concentration of 200 mg/l of drinking-water (stop model). Using a laser dissection technique and biochemical microanalysis, the activity of the lysosomal enzyme alpha-glucosidase was measured in glycogen storage foci emerging early, and in mixed or basophilic cell populations (foci and carcinomas) appearing later during hepatocarcinogenesis. In the liver tissue of normal appearance in both untreated controls and NNM-treated animals a slight gradient of alpha-glucosidase activity was observed leading from relatively high activities in zone 1 to lower activities in zone 3 of the liver lobule. In preneoplastic glycogen storage foci a considerable relative reduction in alpha-glucosidase activity was detected, suggesting that a decrease in the hydrolytic glycogen degradation contributes to the disturbance in phosphorylytic glycogen breakdown observed earlier in the majority of the glycogenotic foci. In contrast with glycogen storage foci, mixed and basophilic cell foci and particularly hepatocellular carcinomas showed a marked increase in alpha-glucosidase activity compared with that of normal liver tissue. The gradual enhancement in enzyme activity appeared to be closely related to the reduction in glycogen initially stored in excess during the later stages of hepatocarcinogenesis. The results support the concept that a fundamental shift in carbohydrate metabolism is characteristic of neoplastic transformation of hepatocytes. PMID- 2569787 TI - Ultrastructure of neurofibrillary tangles in Alzheimer's disease. AB - The ultrastructure of neurofibrillary tangles (NFT) was examined by electron microscopy. The fibrils of NFT seemed to consists of about eight protofilaments consisting of globular subunits; these protofilaments were helically wound in a longitudinal direction. The fibrils of NFT had hollow structures at their centers surrounded by the eight globular subunits. The subunits were tightly connected in the narrow parts of the fibril, but more loosely connected in the wider parts. From these findings, it seemed that the fibrils of NFT consist of a twisted tubule having periodical constrictions and is made up of eight helically wound protofilaments, forming globular subunits. PMID- 2569786 TI - Immunohistochemical demonstration of lysozyme and lactoferrin in salivary pleomorphic adenomas. AB - Immunohistochemical identification of lysozyme and lactoferrin was made in salivary pleomorphic adenomas (147 cases) and the staining patterns were evaluated with respect to the histological features and histogenesis. In normal salivary glands, the intercalated duct cells gave positive staining for lysozyme in major glands, and serous acinar cells, demilune cells, and interlobular duct cells were positive in minor glands. Lactoferrin staining was irregularly positive in serous cells and ductal epithelium. In pleomorphic adenomas, the reaction for lysozyme was positive in 14% (21/147) of the cases, and was confined to luminal cells of tubulo-ductal structures. Lactoferrin in pleomorphic adenomas was distributed in luminal tumor cells (51%; 75/147), in outer tumor cells (3%; 4/147), and in both luminal and outer tumor cells (5%; 7/147) in tubulo-ductal structures; it was also detected in plasmacytoid myoepithelial cells (5%, 8/147). However, modified myoepithelial cells and other types of neoplastic myoepithelial participants were negative for lactoferrin staining. The occurrence of both lysozyme and lactoferrin in salivary pleomorphic adenomas suggests their participation in the local defense mechanism in the tumor. PMID- 2569788 TI - [The possible effect of a ketotifen preparation on beta-receptor blockade]. AB - Parallel clinico-pharmacological studies were carried out of ketotifen action in patients with atopic bronchial asthma and in an experimental model of beta adrenergic blockade in guinea pigs. The results revealed that ketotifen acts on the beta-adrenergic blockade in the patients with atopic bronchial asthma and in the experimental model in guinea pigs. The ketotifen action on beta-adrenergic blockade resembles that of glucocorticosteroids in efficiency. PMID- 2569789 TI - [Chronic hyposomnia]. AB - The problem area of chronic insomnia will be discussed in view of diagnostic, causal and therapeutic aspects. A duration of an insomnia more than 3 weeks is regarded as chronic, it demands a detailed anamnestic interview as well as physical examinations. The chronic intake of hypnotics rather often causes insomnia. The reversible influence of chronic intake of hypnotics on polygraphic sleep parameters are shown during withdrawal, which was carried out stepwise. Alternative therapeutic strategies are discussed. PMID- 2569790 TI - [Sleep disorders in the aged and their treatment]. AB - A shortening of sleep duration is going along with aging. Several somatic diseases and different psychiatric disorders, which increasingly occur in old people cause sleep disorders. Thus patients suffering from disturbed sleep must be examined carefully before treatment. Benzodiazepines and Clomethiazol are potent and save drugs for sleep disorders in the aged. Antidepressants and neuroleptics should be applicated only for sleep disorders caused by underlying psychiatric diseases. PMID- 2569791 TI - [Clinical aspects of hypnotics]. AB - Knowledge of benefits and risks of these substances are important for a low-risk treatment with hypnotics. Different questions occur, such as short- oder long acting hypnotics? Disturbing interactions? Duration of therapy? Risks of amnesia? Insomnia of old people? Alternatives to Benzodiazepin-Hypnotics? In each case the application of these hypnotics should follow along the "3 S Line": straight indication, small dose, short application. There does not exist an universal principle of therapy for people who suffer from insomnia. This serious disease is often very agonizing for them. As a pharmacologic instrument there is yet no alternative to Benzodiazepines. PMID- 2569792 TI - Changing patterns of psychotropic drug use in the elderly: a five-year update. AB - Psychotropic drug use was evaluated in 2022 ambulatory elderly subjects in 1978 80 and again in 1984-86. Use of hypnotic drugs declined from 8.5 percent (n = 3234) in 1978-80 to 6.3 percent (n = 2681) in 1984-86 (p less than 0.01). Use of the long-acting hypnotic flurazepam decreased (p less than 0.01) and use of two short-acting drugs, triazolam and temazepam, increased. Prescribing of long half life benzodiazepines, such as diazepam (p less than 0.01) and chlordiazepoxide, clorazepate, halazepam, and prazepam as a group (p less than 0.01) decreased as well as the use of nearly all products containing barbiturates (p less than 0.01). PMID- 2569794 TI - [Welcome to the first meeting. Health and nursing care meeting]. PMID- 2569795 TI - [Sedative and tranquilizing drugs--dosage determination with great care for older patients]. PMID- 2569796 TI - [RFHL (National Organization for Help to Drug Abusers): individuals in the health care professions seek us out with their dependencies]. PMID- 2569793 TI - Intra- and extracellular amino acid concentrations in portacaval-shunted rabbits. Role of hyperammonemia and effects of branched-chain amino acid-enriched parenteral nutrition. AB - Intra- and extracellular amino acid concentrations were measured in rabbits in order to elucidate the possible role of hyperammonemia in lowering the postabsorptive plasma levels of branched-chain amino acids (BCAA) and to assess the effects of BCAA-enriched total parenteral nutrition (TPN) on the amino acid pattern of muscle. The pathophysiological part of this paper deals with portacaval anastomosis (PCA) and is aimed at substantiating or rejecting our hypothesis that excessive ammonia-by stimulating glutamine synthesis-reduces the intracellular glutamate pool which is then restored, at least in part, by an intensified BCAA degradation. Regarding infusion therapy, we were mainly interested in whether an amino acid solution adapted to the metabolism in liver cirrhosis causes an accumulation of BCAA in muscle or modifies the intracellular content of glutamate and glutamine. Eighteen rabbits did not undergo surgery and served as controls (group A), while 30 were given a portacaval end-to-side anastomosis (group B). Two weeks after creating the PCA, venous blood samples were taken and muscle biopsies (Bergstrom's technique) were performed postabsorptively. An 18-h TPN was then started, the regimen administered included dextrose, fat and, in addition, either a conventional (group B1, n = 15) or an adapted amino acid solution (group B2, n = 15). We obtained second blood specimens and muscle biopsies at the end of the infusion period. With the control animals, the same time schedule for blood sampling and muscle biopsies was followed. Fourteen days after the operation, the PCA rabbits displayed a mean plasma ammonia level 5.1 times higher than that measured in the controls (p less than or equal to 0.001). Conventional blood chemistry did not reveal any impairment of liver cell integrity or over-all hepatic function, whereas the nutritional state of the shunted animals worsened, as indicated by body weight and biochemical variables. Since in the PCA rabbits, the total amino acid pools of muscle and plasma were seen to be increased and decreased, respectively, the results concerning the individual amino acids are given in terms of both the absolute and percentage values, the latter more often revealing high levels of statistical significance. PCA induced a marked rise in the intra- and extracellular concentrations of glutamine, while the values of glutamate and alanine showed a decline in muscle and plasma. The extracellular levels of methionine, phenylalanine, and tyrosine were raised, while those of the BCAA were diminished.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2569797 TI - ["I was stupefied and lived at the fringe of life"]. PMID- 2569798 TI - [200,000 are dependent on tranquilizing agents--6 beds for detoxification are available in St Goran]. PMID- 2569800 TI - Alteration of T cell subset pattern among in vitro M. leprae-stimulated blood lymphocytes of leprosy patients. PMID- 2569799 TI - [The behavior of serum "liver enzyme" activity in children with chronic renal failure, hemodialysis and kidney transplantation]. AB - Liver enzymes were measured in 24 children with different degrees of renal insufficiency, in 6 children treated by chronic hemodialysis and in 13 children after kidney transplantation. The hemodialyzed and transplanted patients have the highest ALAT activity, independently of the presence of a liver infection. The AP activity in these patient groups were not different from the reference value. The activity of ALAT, GGT and CHE, respectively were most pathologic in the hemodialysis group. The enzyme activities have not been influenced by a single dialysis and a 6-month dialysis course. In uremic and chronic hemodialyzed patients the enzyme combination of ALAT, GGT and CHE has been recommended in the diagnosis and follow-up control of a liver cell damage. In the evaluation of enzyme activities in comparison with reference values of healthy volunteers an enzyme inhibition by the uremic serum should considered. PMID- 2569801 TI - A study of the relatedness of Mycobacterium leprae isolates using restriction fragment length polymorphism analysis. AB - The inability to cultivate Mycobacterium leprae in vitro has severely hampered comprehensive phenotypic analysis of individual isolates, leaving unanswered the question of the relatedness of these isolates. Since the nucleotide sequence of a bacterial chromosome is its "genetic fingerprint", we have employed the use of restriction fragment length polymorphism (RFLP) analysis of chromosomal DNA of M. leprae isolates to assess the relatedness among these isolates. DNA of M. leprae was harvested from infected armadillo tissue originally inoculated with bacilli from lepromatous lesions of human patients from geographically distinct regions of the world. Restriction endonuclease (EcoRI, PstI, and PvuI) digests of chromosomal DNA were analysed using DNA probes encoding all or part of the 28 kDa, 65-kDa and 70-kDa proteins of M. leprae. Comparison of the resultant autoradiographs showed that the RFLP patterns were all identical indicating that these isolates contained no polymorphism with respect to the restriction endonuclease sites analysed. These results indicated that the M. leprae isolates tested in this study were indistinguishable at the genotypic level, suggesting the possibility of homogeneity among members of the species, M. leprae. PMID- 2569802 TI - The genetic relationship between Mycobacterium paratuberculosis and the M. avium complex. PMID- 2569803 TI - Clinical characteristics, health beliefs and compliance with maintenance treatment: a comparison between regular and irregular attenders at a depot clinic. AB - This study investigated the relationship between clinical characteristics, health beliefs and compliance with depot neuroleptic maintenance treatment in schizophrenic patients by comparing a group of 40 irregular clinic attenders with another group of regular attenders matched by age and sex. While the 2 groups showed no difference in drug-induced extrapyramidal effects, the former were significantly more often depressed than the latter. Test-retest reliability coefficients of a 4-section health belief questionnaire showed marked variation in individual items. The 2 groups showed no difference in their perception of past effect of illness, likelihood of a relapse if maintenance treatment was stopped and effects of maintenance treatment. PMID- 2569804 TI - Successful treatment with clonazepam for neuroleptic-induced akathisia. AB - Twenty-one patients with severe neuroleptic-induced akathisia resistant to antiparkinsonian medications were treated with oral clonazepam in an open trial. All the patients reported subjective improvement. We suggest that clonazepam is effective in managing neuroleptic-induced akathisia. PMID- 2569805 TI - Effect of viral infection of Fc receptors and immunologically nonspecific receptors of blood polymorphonuclear leukocytes: an in vitro model. AB - I describe the effect on phagocytic activity of the migration of bovine blood polymorphonuclear leukocytes (PMNL) through collagen and a monolayer of PK-15 cells in vitro infected with rotaviruses (strain OSU, RFC-17, SA-11), a picornavirus (strain SVDV), bovine herpesvirus (BHV-1) and equine herpesvirus (EHV-1). The PMNL were examined before and after their migration as follows: (1) for EA rosette formation with sensitized sheep erythrocytes, (2) for phagocytic activity mediated through Fc receptors (FcRs), (3) for phagocytic activity mediated through immunologically nonspecific receptors (INsRs). Random migration of PMNL through the infected monolayer to RPMI-1640 medium containing or lacking chemotactic agents (zymosan activated serum--Act. serum) and/or containing control agents (lipopolysaccharide--LPS or formyl-L-methionyl-L-leucyl-L phenylalanine--fMLP) decreased the percentage of rosettes forming PMNL and the phagocytosis mediated by FcRs and INsRs. The results obtained suggest that the viruses used in this study independently of their biological properties exerted a suppressive action on the phagocytic activity mediated by FcRs and INsRs. PMID- 2569807 TI - Hybridoma cell lines secreting monoclonal antibodies to foot-and-mouth disease virus type Asia-1. AB - Various immunizing regimens, cell culture requirements and cell fusion conditions were examined for efficient production of hybridomas secreting anti-foot-and mouth disease virus (FMDV) antibodies. A highly sensitive streptavidin-biotin based enzyme-linked immunosorbent assay (ELISA) was used for screening of hybridomas for specific antibody production as well as for determining the serotype specificity of the antibodies. Six hybridoma cell lines generating antibodies to FMDV type Asia-1 (vaccine strain 63/72) were produced by fusion of SP2/0 mouse myeloma cells and spleen cells from mice immunized with the inactivated viral antigen. The monoclonal antibodies (MoAb) from four producer clones reacted in ELISA specifically with the intact virus antigen of type Asia-1 without any cross-reactivity with strains of other virus types 0, A and C. Two other clones positive for anti-Asia-1 virus antibodies in ELISA cross-reacted with type C virus strain. The MoAb from three of the four Asia-1 virus type specific clones neutralized the infectivity of the immunizing viral strain. One neutralizing MoAb reacted with the separated VP1 from the immunizing viral strain in immunoblotting. PMID- 2569806 TI - A radioimmunoassay detecting the bovine leukaemia virus transmembrane protein gp30 and anti-gp30 antibodies in the serum of cattle. AB - By means of SDS PAGE we isolated from virus-infected foetal lamb kidney (FLK) cells a relatively homogenous envelope transmembrane protein gp30 of bovine leukaemia virus (BLV). As shown by a partial sequence analysis of the N-terminus of this protein, our gp30 preparation contained only traces (less than 5%) of p24 gag protein: Rabbit anti-gp30 serum did not cross react with the BLV proteins gp51, p12, p15(1), p15(2), and p10 but reacted weakly with the p24 polypeptide. 125I-labelled gp30 (chloramine-T) was precipitated with the serum of BLV-infected cattle. Nonlabelled preparation of gp30 competitively inhibited the reaction of 125I-labelled gp30 with the natural antibodies. We investigated 193 cattle sera by liquid phase radioimmunoassays (RIA) using 125I-gp30, gp51 and p24 antigens. Sixteen noninfected cattle sera were negative in all tests. The 177 serum samples of BLV-infected animals were examined to the diagnostic value of the three tests. Of these, 175 were positive in gp51 RIA, 172 in p24 RIA and 164 in gp30 RIA. In all three tests, 159 sera were positive while 18 sera, mostly coming from animals with normal leukocyte counts, were positive only either with gp51 or p24, or were double positive with either gp51/p24 or gp51/gp30. We conclude that the gp51 RIA is superior to both the gp30 and the p24 RIA and that the gp30 RIA will be useful for investigating the role of gp30 in virus pathogenicity. PMID- 2569808 TI - Action of rimantadine on the structure of influenza A virus haemagglutinin. AB - Rimantadine prevents the conformational changes of influenza virus haemagglutinin (HA) caused by acid pH and the acquisition of sensitivity to trypsin, protects the haemolytic activity from inactivation and prevents the morphological changes of HA spikes on the virus surface. PMID- 2569809 TI - Expression of proteins immunologically related to murine mammary tumour virus (MMTV) core proteins in the cells of breast cancer continuous lines MCF-7, T47D, MDA-231 and cells from human milk. AB - Expression of antigens immunologically related to the gag gene products of murine mammary tumour virus (MMTV) in continuous cell lines MCF-7, T47D and MDA-231 was followed by indirect enzyme immunoassay (EIA). Some cells of the 2 clonal lines derived from MCF-7 and T47D cells contained MMTV antigens, but these were not detected in MDA-231 cells and in epithelial cells from the milk of healthy women. The expression of antigens related to MMTV gag proteins correlated with the expression of proteins immunologically related to MMTV gp52. Cells positive for antigens related to p27 were also found when healthy women sera containing antibodies against MMTV p27 were examined. Before testing, the donor sera containing antibodies against MMTV protein p27 were absorbed to the negative MCF 7 cell clone and their antibody activity to MMTV p27 was also tested by immunoblotting. PMID- 2569810 TI - Sensitivity and specificity of a modified agar gel precipitation test and its application to the diagnosis of enzootic bovine leukosis. AB - Sensitivity and specificity of a modified agar gel immuno-diffusion (AGID) test for the diagnosis of enzootic bovine leukosis was compared in 609 sera using the standard method, the modified test, and ELISA. The modifications concerned the composition of agar gel (1.8% Bacto-agar Difco), addition of polyethylene glycol (PEG 6000, 4%) and enlarging the diameters of the wells from 6 to 10 mm for sera. The change in well diameters and the addition of PEG 6000 increased the sensitivity of the test by about 60-100%, favourably influenced the proper interpretation of the results, especially with weak positive sera. ELISA proved to be more sensitive either than the modified (by 4.1%) or standard AGID (by 5.25%) tests. The modified AGID test may find wide application in laboratory practice, especially when a more sensitive technique is not available. PMID- 2569811 TI - Detection and analysis of antibodies against retrovirus determinants in the sera of haematologic patients. AB - A panel of 11 retroviruses including animal retroviruses types B, C, D and human retroviruses HTLV-I and HTLV-III was used as antigen for detection of antibodies in 265 sera of patients suffering from haematologic and autoimmune diseases and in 98 control sera. In contrast to the control sera, the 39 patients sera contained antibodies against internal proteins of animal retroviruses types B and C. No antibodies against human retroviruses proteins or type D retrovirus core proteins were detected. The antibodies from positive sera immunoprecipitated the RNA from polyribosomal fraction of R-MuLV infected spleen cells which also showed positive hybridization with a specific cDNA probe. The antibodies in patients' sera were directed mainly against RNA-binding proteins p15 of type C or p14 type B virus. The retrovirus antigenic determinants were expressed also in human embryo haemopoietic cells. The possible interpretation of antibody formation against the determinants of animal retroviruses in humans is discussed. PMID- 2569812 TI - Effects of norakin on respiratory syncytial virus in tissue culture and in mice. AB - Norakin at 1 microgram/ml inhibits the reproduction of respiratory syncytial virus (RSV) in Vero cells to 50% and at 5 micrograms/ml to 90%. The development of lung lesions in RSV-infected BALB/c mice was suppressed by 70% when the animals were treated with two doses (25 mg/kg each) of norakin, 30 min before and 4 hr post infection. (p.i.), respectively. PMID- 2569813 TI - Development of local morphological changes after intradermal inoculation of Q fever chemovaccine. AB - Development of morphological changes was studied in the site of intradermal inoculation of Q-fever chemovaccine. The chemovaccine was obtained by trichloroacetic acid extraction from Coxiella burnetii, strain Nine mile, phase I cells. The effect of two vaccine doses (0.2 mg and 1.0 mg) was compared in two groups of guinea pigs (each consisting of 21 animals). After inoculation of 1.0 mg vaccine abscesses formed in the dermis and in subcutaneous tissue along with dystrophic changes of skeletal muscles. Reparation of these lesions progressed sufficiently slowly and was not completed before 60 days after vaccine inoculation. The regeneration of skeletal muscles was of budding type. After inoculation of 0.2 mg of chemovaccine minimal lesions occurred in the subcutaneous tissue only. However, already within 48 hr this vaccine dose evoked a strong reparation response. Interesting was the finding of Kurloff cells in the haemorrhagic foci. The results suggest that the low-reactogenic chemovaccine dose of 0.2 mg might be sufficiently immunogenic after subcutaneous administration. PMID- 2569814 TI - Comparison of protein and lipopolysaccharide patterns of several Coxiella burnetii strains in phase I. AB - Electrophoresis in polyacrylamide gel gradients in the presence of sodium dodecyl sulphate revealed at least 18 identical protein bands in eight strains of Coxiella burnetii. By staining with Coomassie Brilliant Blue R-250 it was possible to visualize clearly at least 40 proteins. The protein pattern showed the greatest variability in the region from 27 to 18.5 kD. Strains S and Priscilla differed from the other strains also in proteins smaller than 18.5 kD. The lipopolysaccharide pattern obtained by the same technique consisted of from 6 to 12 various bands in the region from 23.5 to 11.8 kD. The protein and lipopolysaccharide patterns of four strains isolated from ticks did not significantly differ from those of strains Henzerling and L-35 isolated from men. PMID- 2569815 TI - Effect of storage and 60Co irradiation on the antiviral activity and subtype pattern of native human interferon-alpha preparations. AB - The stability of native human leukocyte IFN-alpha was investigated after one year storage at different temperatures, or after sterilization with 60Co-irradiation by determining the total antiviral activity of samples. Inactivation of VSV and Aujeszky virions by 60Co-irradiation was directly related to the radiolysis of samples, indicating the uselessness of this procedure for sterilization of IFN alpha preparations. The presence and proportion of subtypes in the stored or irradiated preparations (with 50 and 25-70% inactivation, respectively) was analysed by chromatofocusing, comparing their patterns with that of the untreated controls. A logarithmic correlation was found between the pI values and temperature/irradiation sensitivity of subtypes. PMID- 2569816 TI - A laboratory study of age-related varicella incidence and prevalence in the Czech Socialist Republic. AB - During 1972-1981, ten representative population samples totalling 2,916 individuals were tested for antibodies to varicella-zoster virus (VZV) by an indirect-haemagglutination assay (IHA). Statistical analysis of the results provided estimates of age-related varicella prevalence and incidence rates. It transpired that 45% of the child population had encountered varicella at preschool age and another 45% during the attendance of school. Adult seropositivity rates amounted to 97.5-100%. The highest varicella incidence was observed in the 4-year age interval of 2-6 years. For control, additional 324 and 297 individuals were tested by RIA and ELISA IgG, respectively; there was good correlation of results. For the period 1970-1985 varicella morbidity was also studied from notification data. As far as the authors are aware, the present investigation performed on a total of 3,537 subjects furnishes the most comprehensive information on nation-wide varicella incidence and prevalence obtained by laboratory methods. PMID- 2569817 TI - Stabilization of the attenuated poliovirus type 3 vaccine strain by sucrose. AB - Highly temperature-sensitive and attenuated poliovirus type 3, strain Leon 12a1b, was stabilized at 37 degrees C by 15% sucrose without 1 mol/1 MgCl2. The finding will contribute to poliovaccination in tropical countries. PMID- 2569818 TI - HIV-2 infection in Czechoslovakia. PMID- 2569819 TI - Changes of brain gangliosides in rat infected with Japanese encephalitis virus. PMID- 2569820 TI - Preparation and biochemical characterization of Lassa virus subvirion fractions. AB - 14C-labelled Lassa virus was purified by isopycnic centrifugation and split to subvirion fractions. The purified virus was treated with nonionic detergents Nonidet P-40 (NP-40) and with octylglycoside. After ultracentrifugation in urographin density gradient, two subvirion fractions with buoyant density of 1.24 1.26 and 1.08-1.10 g/cm3 were obtained. The first fraction corresponded to the nucleocapsid of Lassa virus: it contained a protein with molecular mass of 60 kDa, the L and S segments of the genomic RNA. The second one contained a protein with molecular mass of 48 kDa and represented, apparently, envelope fraction of virus particles. PMID- 2569821 TI - Clinical settings for selective alpha-adrenergic receptor inhibition: rationale and management strategies. Proceedings of a symposium. Naples, Florida, October 13 to 16, 1988. PMID- 2569822 TI - Clinical pharmacotherapeutics of doxazosin. AB - Doxazosin is the latest in a series of highly selective postsynaptic alpha 1 adrenoceptor inhibitors. It is readily absorbed, with high bioavailability and a relatively long plasma half-life, neither of which property is influenced by age. This accounts for the prolonged pharmacologic activity of doxazosin following a single oral dose. Its prime pharmacodynamic activity resides in its ability to counter sympathetic vasoconstriction of the systemic arteriolar resistance vessels and venous capacitance system, which enables the drug to target the major pathophysiologic abnormality in hypertension, i.e., the generalized systemic arteriolar constriction. The widespread vasodilation induced by doxazosin relieves both cardiac preload and afterload and, consequently, reduces left ventricular wall stress and myocardial oxygen consumption. In hypertension, doxazosin reduces blood pressure both at rest and during exercise by reduction of systemic vascular resistance without precipitating substantial reflex cardiac stimulation. The effects are maximal on the standing blood pressure between two and four hours after ingestion; due to doxazosin's relatively slow absorption, postural hypotension is infrequent. Its antihypertensive activity is maintained over 24 hours following a single oral dose, and the optimal dose range is 2 to 8 mg once daily. The antihypertensive efficacy of doxazosin has been shown to be comparable with that of other alpha-adrenoceptor inhibitors, beta-blocking drugs, diuretics, calcium antagonists, and angiotensin-converting enzyme inhibitors. In contrast to other conventional antihypertensive drugs, a unique feature of alpha adrenoceptor-inhibiting drugs, including doxazosin, is their ability to reduce the plasma concentrations of triglycerides, total cholesterol, and low-density lipoprotein cholesterol and to increase high-density lipoprotein cholesterol concentration. This contrasts with the opposite effect on lipid levels induced by hydrochlorothiazide and atenolol seen in comparative studies. Side effects show no predilection for any organ system, and the overall incidence of such effects compares well with those of other commonly used antihypertensive drugs. This unique combination of antihypertensive efficacy and favorable effect on blood lipid levels indicates that once-daily treatment with doxazosin holds considerable promise in the treatment of hypertension, both from the point of view of its antihypertensive efficacy and also from its primary preventative potential. PMID- 2569823 TI - Pharmacologic basis for the use of doxazosin in the treatment of essential hypertension. AB - The drug treatment of mild hypertension has been shown to afford protection against fatal and nonfatal strokes, congestive heart failure, progression to more severe levels of hypertension, and all-cause mortality, but not against the complications of coronary artery disease. The lack of benefit against coronary artery disease may result from failure to reduce other risk factors or because the drugs employed increased coronary risk. It can be taken as axiomatic that effective preventive antihypertensive therapy is more likely with drugs with mechanisms and sites of action that are focused on the underlying pathophysiology than with drugs that lower blood pressure by means unrelated to the hypertensive process. Adrenergic predominance plays a major role in the initiation and maintenance of essential hypertension and, consequently, the alpha-adrenergic receptor inhibitors were among the first substances to receive serious consideration as antihypertensive agents. However, since these drugs are nonselective, feedback control of transmitter norepinephrine was lost and, consequently, the clinical expectations of the early alpha-adrenergic receptor inhibitors in the treatment of high blood pressure were not fulfilled. The discovery of selective postjunctional alpha 1-adrenergic-receptor inhibitors, such as prazosin and doxazosin, which preserve feedback control of transmitter norepinephrine release, was the crucially important step in the development of specific drugs to combat the hyperactivity of adrenergic vasoconstrictor nerves in hypertension. These drugs have been shown to normalize hemodynamics in hypertensive patients. They lower blood pressure through a reduction in peripheral resistance at rest and during exercise, independent of changes in heart rate and blood pressure, with minimal reflex activation or tolerance development. Alpha 1-adrenergic-receptor inhibitors, such as prazosin and doxazosin, represent an attractive choice for initial therapy in all grades of hypertension and are especially appropriate in hypertensive patients with congestive heart failure, asthma and chronic obstructive airways disease, renal impairment, diabetes mellitus, hyperlipidemia, benign prostatic hyperplasia, or gout, and in those involved in vigorous work, sports, or exercise. There are no known contraindications to these drugs, except in patients who are sensitive to quinazolines. PMID- 2569824 TI - Selective alpha 1-adrenergic blockade, lipids, and coronary heart disease risk. Considerations in the treatment of mild hypertension. AB - The importance of hypertension and hyperlipidemia as independent and interactive risk factors for the development of premature cardiovascular disease, and particularly for the development of atherosclerotic coronary heart disease, is becoming increasingly apparent both from epidemiologic data and from therapeutic trials. Nevertheless, therapeutic trials of patients with mild hypertension have not demonstrated benefits from lowering arterial pressure, in terms of reduced mortality rates from coronary events. This may, in part, be due to the fact that many of the antihypertensive agents used in these trials adversely influence lipid and lipoprotein levels. Thus, agents that are lipid-neutral or that favorably influence the lipid profile, such as selective alpha 1-inhibitors, are receiving increasing attention for the treatment of mild hypertension. Recent insights into these issues are considered. PMID- 2569825 TI - Review of the effects of doxazosin, a new selective alpha 1-adrenergic inhibitor, on lipoproteins in patients with essential hypertension. AB - Control of high blood pressure has failed to reduce the risk of atherosclerotic coronary heart disease (CHD). Hypercholesterolemia, which is common among hypertensive patients, cigarette smoking, and hypertension are the major risk factors for CHD. To minimize CHD risk, elevated blood pressure and atherogenic lipid levels should be lowered, but various antihypertensive agents appear to adversely affect lipid levels, actually precluding the CHD risk reduction expected from blood pressure control. Doxazosin, a once-daily, long-acting, alpha 1-adrenergic inhibitor, not only is effective therapy for essential hypertension but also has a favorable impact on lipids. During controlled studies of doxazosin's antihypertensive efficacy, the following blood lipid levels were measured: total cholesterol, total triglycerides, high-density lipoprotein (HDL) cholesterol (including HDL2 and HDL3), low-density lipoprotein (LDL) cholesterol, very low-density lipoprotein cholesterol, and apoproteins (apos) AI and B. Results showed total cholesterol (-0.8 to -8.9 percent), total triglycerides ( 5.0 to -17.4 percent), and LDL (-9.0 to -16.9 percent) were reduced. The positive prognostic indicators, HDL (+0.7 to +13.0 percent) and HDL:total cholesterol ratio (+3.1 to +26.3 percent), were increased. Apo B decreased, but apo AI remained unchanged. In these hypertension studies, doxazosin has favorably reduced two major CHD risk factors. As part of a new, long-term, controlled, multicenter trial, the prospective benefits of these risk factor reductions on CHD morbidity and mortality will be quantified for doxazosin and other antihypertensive agents. PMID- 2569826 TI - Ptosis of eyelids, strabismus, diastasis recti, hip defect, cryptorchidism, and developmental delay in two sibs. AB - We report a distinct syndrome of eyelid ptosis, convergent strabismus, abdominal muscle defect, hip dislocation, cryptorchidism and developmental delay in two brothers. Consanguinity in their parents suggests autosomal recessive inheritance. PMID- 2569827 TI - Prenatal diagnosis of cystic fibrosis by using linked DNA markers in 138 pregnancies at 1-in-4 risk. AB - Prenatal diagnosis was carried out in 138 pregnancies at 1-in-4 risk for cystic fibrosis (CF) by using closely linked DNA markers, including XV-2c and KM-19. In fully informative families, 25 of 123 (20%) fetuses were predicted to be affected; 16 of these 25 pregnancies were terminated and 9 were continued. Postnatal sweat tests are completed in 42 cases; the diagnoses were confirmed in 4 of 4 infants predicted to be affected and in 37 of 38 infants predicted to be unaffected. One infant predicted to be a carrier had an abnormal sweat test after birth, but the mother also had an abnormal sweat test, and there was no evidence of an error in linkage analysis. The data indicate that prenatal diagnosis using linkage analysis is fully informative in most families and is highly reliable with either chorionic villus sampling or amniocentesis. Although outcome data are available on only 42 pregnancies, based on our experience, on general principles of linkage analysis, and on the tight linkage of the known DNA markers with CF, we recommend that DNA analysis replace microvillar intestinal enzyme analysis for 1-in-4 risk pregnancies when DNA is available from the propositus. PMID- 2569828 TI - Plasma membrane-specific deficiency in cardiac beta-adrenergic receptor in streptozocin-diabetic rats. AB - Cell surface [3H]CGP 12177 binding sites in 10-wk streptozocin-diabetic rats decreased by 41% (P less than 0.01) compared with that in the control rats. In contrast, there was no difference in the total cell receptor concentration between the control and the diabetic rats, which was measured by hydrophobic antagonist [125I]-iodocyanopindolol binding. Forty-eight-hour in vivo insulin treatment significantly (P less than 0.05) increased cell surface beta-adrenergic receptor concentration by 37% above that in diabetic rats without any change in total receptor concentration in the cells. However in vitro treatment of 8 nM insulin, 33 mM glucose, or 10 mM 3-hydroxybutyrate for 2 h showed no effect on [3H]CGP 12177 binding. In contrast, 10 microM isoproterenol-dependent decrease and the recovery of cell surface receptors after the removal of the agonist were significantly (P less than 0.01) impaired in diabetic rats compared with those of control rats. These results indicate that only cell surface beta-adrenergic receptors decrease in diabetic rats, which may be associated with abnormalities in the receptor distribution. The decrease in cell surface receptor number closely associates with the diabetic state and is reversed by the short-term insulin treatment. PMID- 2569829 TI - Epinephrine's effect on metabolic rate is independent of changes in plasma insulin or glucagon. AB - Epinephrine's effect to increase metabolic rate is accompanied by changes in the plasma concentrations of insulin, glucagon, and metabolic substrates. Because both glucagon and insulin have been reported to affect thermogenesis, these hormones might contribute to or modify the thermogenic response to epinephrine. To determine if the epinephrine-induced increase in metabolic rate is secondary to changes in glucagon or insulin or to changes in the fuels modulated by these hormones, metabolic rate was measured by indirect calorimetry in five normal weight post-absorptive young men on three occasions: study A, an intravenous epinephrine infusion alone; study B, a 4-h "islet clamp" consisting of somatostatin infusion with basal insulin and glucagon replacement; and study C, an intravenous epinephrine infusion combined with the islet clamp. A 1-h base line period preceded 2 h of epinephrine infusion. During the 4-h islet clamp (study B), metabolic rate and plasma concentrations of epinephrine, insulin, glucagon, and glucose remained unchanged. During the infusion of epinephrine alone (study A), metabolic rate and concentrations of glucagon, free fatty acids, and C-peptide increased as expected. Also as expected, the glycemic response to epinephrine infusion was much larger when insulin and glucagon levels were fixed with the islet clamp (study C). In contrast, the metabolic rate and the free fatty acid concentration responded similarly to epinephrine infusion when insulin and glucagon were fixed (study C) and when they were changing (study A). We conclude that epinephrine increases metabolic rate independently of physiological changes in plasma glucagon or insulin or the circulating fuels they modulate. PMID- 2569830 TI - Induction of neurally mediated NaHCO3 secretion by luminal distension in rat ileum. AB - Distension induces secretion in the intact intestine, but the mechanism of this secretory process remains unresolved. We sought to characterize the effect of intraluminal pressures below 20 cmH2O on water and electrolyte movements in the rat ileum and the subsequent effects of two cholinergic antagonists, hexamethonium and atropine. An increase in intraluminal pressure from 3.0 to 12.5 cmH2O led to inhibition of net H2O and Na absorption and stimulation of HCO3 secretion. However, there was no significant change in Cl absorption. Secretion occurred in the absence of changes in tissue wet weight, intercellular fluid accumulation, villus tip erosions, or mannitol flux. Alterations in fluid and electrolyte absorption were prevented by the intra-arterial administration of hexamethonium (10 mg/kg). Atropine (0.5 mg/kg) had no effect. Our studies demonstrate that distension induces the secretion of Na and HCO3 by the intact ileum. This secretory process may represent a neurally rather than passively mediated mechanism. PMID- 2569831 TI - Characterization of cholinergic receptors mediating pepsinogen secretion from chief cells. AB - By use of a highly enriched preparation of chief cells (greater than 90% pure) prepared from guinea pig stomach, the cholinergic receptors regulating pepsinogen secretion were studied. Each of five different muscarinic cholinergic agonists, but not the nicotinic cholinergic agonist nicotine, stimulated pepsinogen release. Carbamylcholine-stimulated pepsinogen release was inhibited by each of seven different cholinergic receptor antagonists with relative potencies of N methylscopolamine greater than scopolamine = 4-diphenylacetoxy-N-methylpiperidine methiodide = atropine much greater than pirenzepine greater than AF-DX-116 much greater than tetraethylammonium. Binding of [N-methyl-3H]scopolamine ([3H]NMS) was time and temperature dependent, reversible, saturable, and specific. Analysis of [3H]NMS binding demonstrated a Kd of 1.3 nM for NMS with a binding capacity of 61 fmol/mg protein or 5,920 sites/chief cell. With the agonists carbamylcholine, acetylcholine, or muscarine, the receptor population could be divided into two classes of receptor sites: a class with high affinity (Kd, 12-53 microM) representing 73% of the binding sites and a class with low affinity (Kd, 2-5 mM) representing 27% of the binding sites. Each of the antagonists had equal affinity for both receptor populations. There was a close correlation between the ability of antagonists to inhibit [3H]NMS binding and carbamylcholine-stimulated pepsinogen release. Each agonist was 29- to 63-fold more potent at stimulating pepsinogen release than interacting with high-affinity receptors and 2,000- to 11,000-fold more potent than interacting with low-affinity receptors, suggesting spare receptors. Studies with the alkylating agent, propylbenzilylcholine mustard, demonstrated that there was a receptor reserve of 50-80%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569832 TI - Somatostatin receptors on canine fundic D-cells: evidence for autocrine regulation of gastric somatostatin. AB - Somatostatin is known to have inhibitory effects on the release and action of a wide variety of gut peptides. Previous studies in vivo have suggested a potential inhibitory role for somatostatin even on its own secretion. To determine whether this autoregulatory effect is the result of a direct action of the peptide on the cell that is responsible for its secretion, we examined the effect of a non immunoreactive but biologically active analogue of somatostatin ([Leu8-D-Trp22 Tyr25]S28) on release of somatostatin-like immunoreactivity (SLI) from isolated canine fundic D-cells. We identified somatostatin binding sites with dissociation constants of 1.2 X 10(-9) and 3.8 X 10(-8) M that coenriched with D-cells. Somatostatin 14, somatostatin 28, and [Leu8-D-Trp22-Tyr25]S28 were equivalent in displacing 125I-[Leu8-D-Trp22-Tyr25]S28 from the binding sites. [Leu8-D-Trp22 Tyr25]S28 inhibited SLI release from D-cells stimulated with (DBcAMP), and pentagastrin. Pertussis toxin pretreatment prevented the inhibitory effects of [Leu8-D-Trp22-Tyr25]S28 on both SLI secretion and cAMP accumulation by on both SLI secretion and cAMP accumulation by D-cells stimulated with epinephrine and forskolin. In contrast, [Leu8-D-Trp22-Tyr25]S28 inhibition of SLI release induced by DBcAMP and pentagastrin was not altered by pertussis toxin. Our data suggest that somatostatin autoregulates its own secretion via specific receptors on D cells. This inhibitory effect is mediated by mechanisms that are both dependent on and independent of pertussis toxin-sensitive inhibitory guanine nucleotide binding proteins. PMID- 2569833 TI - Somatostatin release from isolated ganglia of the myenteric plexus. AB - Somatostatin neurons of the myenteric plexus project caudad exclusively within the plexus synapsing with neurons in the same or other ganglia. Isolated ganglia offer a unique opportunity to examine peptide transmitter release from these interneurons. Ganglia were isolated from the myenteric plexus by sequential enzymatic digestion, centrifugation, and filtration. Single ganglia were harvested by suction and dispersed in polyacrylamide gel. The ganglia were placed in chambers (200 ganglia/chamber) and perfused with Krebs medium at the rate of 1 ml/min. Addition of the nicotinic agonist, dimethylphenylpiperazinium (DMPP), caused a concomitant increase in somatostatin (152%; P less than 0.001) and vasoactive intestinal peptide (VIP) (79%; P less than 0.05) release above basal level that was abolished by hexamethonium. Addition of the VIP antagonist, VIP (10-28), (5 microM), to the perfusate augmented significantly DMPP-induced somatostatin release (440%; P less than 0.05), implying that the concomitant release of VIP attenuates somatostatin release. Addition of VIP (1 microM) to the perfusate abolished somatostatin release, confirming the ability of VIP to inhibit somatostatin release. The study shows that VIP interneurons exert a regulatory feedback control over somatostatin interneurons in the myenteric plexus. PMID- 2569834 TI - Endotoxin increases lung Cu,Zn superoxide dismutase mRNA: O2 raises enzyme synthesis. AB - Administration of endotoxin to adult rats increases lung Cu,Zn superoxide activity after 72 h of exposure to greater than 95% O2. The increased activity is brought about mainly by a faster rate of Cu,Zn superoxide dismutase synthesis; rats treated with endotoxin but not exposed to hyperoxia do not exhibit these findings (Hass, Frank, and Massaro, J. Biol. Chem. 257: 9379-9383, 1982). We now report that 48 h after treatment of adult rats with endotoxin there was a decreased rate of Cu,Zn superoxide dismutase synthesis by lung slices from air- and O2- exposed rats, although, in both groups, the lung concentration of Cu,Zn superoxide dismutase mRNA was increased approximately 45%. Exposure of endotoxin treated rats to greater than 95% O2 or air for an additional 24 h (72 h all told) resulted in continued elevation of Cu,Zn superoxide dismutase mRNA only in lungs of O2- exposed rats. In vitro exposure of lung slices from air-breathing saline- or endotoxin-treated rats to 95% O2 for 6 h led to an increased rate of Cu,Zn superoxide dismutase synthesis only in slices from endotoxin-treated rats. We conclude that endotoxin treatment leads to an increased concentration of Cu,Zn superoxide dismutase mRNA in rat lungs, but a sustained elevation of the mRNA, and its translation into an increased rate of Cu,Zn superoxide dismutase synthesis requires exposure of the lung to hyperoxia. PMID- 2569835 TI - Carrier-mediated transport of tetraethylammonium across rabbit renal basolateral membrane. AB - Mechanisms of tetraethylammonium (TEA) transport were investigated in basolateral membrane (BLM) vesicles from rabbit renal cortex. Preloading vesicles with 10 mM TEA or 1 mM mepiperphenidol stimulated the 15-s uptake of [14C]-TEA compared with control vesicles (258 and 200%, respectively) and produced an overshoot of the equilibrium value (3 and 1.6 times, respectively). In the presence of a K+ gradient, net TEA uptake was also increased (and showed an overshoot of 2-fold) when the membrane potential of vesicles was made interior negative by adding valinomycin. Both TEA-TEA exchange and the voltage-driven net TEA transport were cis-inhibited by other organic cations, and a similar affinity order was found for both transport mechanisms (quinine greater than amiloride greater than morphine greater than mepiperphenidol greater than choline = N1 methylnicotinamide). This data suggests that the same transport protein might be responsible for both phenomena. Other experiments determined that the BLM vesicles had no TEA-H+ exchange, and that contamination of the vesicle population by brush-border membranes was negligible in terms of their contribution to TEA transport. These results demonstrate the presence of an exchange reaction for TEA in the rabbit renal BLM and thus imply carrier-mediated transport of TEA by these membranes. PMID- 2569836 TI - Electrical properties of rabbit early distal convoluted tubule in primary culture. AB - Distal bright convoluted tubules (DCTb) were microdissected from rabbit kidney cortex and cultured in a hormonally defined medium. The quality and the degree of polarization of the growing epithelia were assessed by indirect immunofluorescence studies using a monoclonal antibody raised against the apical membrane of the DCTb in situ. The cultured monolayers had a high hexokinase activity and a low gamma-glutamyl transferase activity compared with cultured proximal convoluted tubules. Adenosine 3',5'-cyclic monophosphate production was stimulated by calcitonin and insensitive to parathyroid hormone, vasopressin, and isoproterenol. Both 20- and 30-day-old cultures developed an apical-negative transepithelial potential of -3.1 and -22.3 mV, respectively. Amiloride reversibly reduced the voltage by 90% only when applied on the apical side of the monolayers. Phenamil (10(-8), 10(-6) M) had the same effect as amiloride. Calcitonin reversibly decreased the transepithelial voltage. These data support the hypothesis that, in the DCTb in primary culture, the transepithelial voltage is due to the presence of Na channels and that calcitonin modulates this transport pathway. PMID- 2569837 TI - Somatostatin stimulates acetylcholine release in the canine heart. AB - Previous reports of somatostatin's atropine-sensitive negative inotropic effect on cardiac function prompted the present studies to characterize the molecular forms and actions of somatostatin in the canine heart. Radioimmunoassay of cardiac extracts revealed concentrations of somatostatin-like immunoreactivity ranging from 0.50 +/- 0.13 pmol/g tissue (means +/- SE, n = 6) in the pulmonary artery to 0.78 +/- 0.23 pmol/g tissue in the right ventricle. On gel filtration of the extracts, two major molecular forms of somatostatin-like immunoreactivity were elicited, the predominant one coeluting with somatostatin-14 and a minor peak corresponding to somatostatin-28. Experiments performed with slices of atrial septum indicated that somatostatin-14 (10(-10) to 10(-7) M) stimulated the release of acetylcholine in a dose-dependent manner. This action of somatostatin 14 was additive with K+-evoked acetylcholine release, unaffected by hexamethonium, and blocked by tetrodotoxin, suggesting mediation via a nonnicotinic postganglionic neural pathway. Our studies lead us to conclude that somatostatin may function as a neurotransmitter in the heart, which exerts its negative inotropic action by promoting the release of acetylcholine. PMID- 2569838 TI - Cardiovascular effects produced by L-glutamate stimulation of the lateral hypothalamic area. AB - L-Glutamate microinjections into the tuberal region of the lateral hypothalamic area (LHAt) caused a fall in blood pressure and heart rate in pentobarbital anesthetized rats. The bradycardia was mediated by both beta-adrenergic and muscarinic mechanisms as demonstrated with pharmacological blockade. The hypotension was due to a decrease in cardiac output, not a decrease in total peripheral resistance. In addition, there was a reduction in coronary blood flow. If heart rate was held constant by pharmacological blockade or by electrical cardiac pacing, L-glutamate stimulation of the LHAt still caused a fall in blood pressure. When the electrically paced model was used, this hypotension was due to a fall in cardiac output. In contrast, with the pharmacological blockade of the heart, the hypotension was due to a decrease in the total peripheral resistance. The cardiac output reduction in the paced condition was not mediated solely by either beta-sympathetic or parasympathetic mechanisms as determined by pharmacological blockade. With heart rate held constant by either drugs or pacing, LHAt stimulation did not alter regional blood flow or resistance in any vascular bed, including the coronary circulation. We conclude that L-glutamate stimulation of the LHAt lowers the cardiac output and heart rate by both parasympathetic and beta-adrenergic mechanisms and elicits hypotension by lowering cardiac output in the naive and electrically paced model. PMID- 2569839 TI - Excitatory and inhibitory effects of gastrin peptides on gastric motility in sheep. AB - In conscious sheep, tetragastrin, pentagastrin, and synthetic human gastrin I, injected either subcutaneously or intravenously in doses of 156-5,200 pmol/kg body wt, inhibited the vagally dependent cyclical motility of the reticulum and rumen, whereas in vitro pentagastrin (10(-12) to 10(-6) M) had no demonstrable inhibitory or excitatory effects on intrinsically active or quiescent muscle of the reticulum, rumen, and omasal leaves. In vitro pentagastrin (10(-18) to 10(-4) M) stimulated quiescent and intrinsically active longitudinal and circular muscles of the body of the omasum and the body and antrum of the abomasum and potentiated contractile responses of antral muscle to electrical stimulation of intramural cholinergic nerves. Responses in the presence of hexamethonium, atropine, and tetrodotoxin indicated that the excitatory effects on mixed nerve muscle preparations of omasal and abomasal tissue were mediated both through stimulation of cholinergic neurones and by direct actions on the muscle. In vitro the ovine stomach shows marked regional differences in both its response and sensitivity to gastrin peptides, and their inhibitory effects on reticuloruminal motility in vivo appear to be other than direct. PMID- 2569840 TI - Baroreceptor activation or glutamate coinjection facilitates depressor responses to ANF microinjection into NTS. AB - As microinjection of atrial natriuretic factor (ANF) into the nucleus of the solitary tract (NTS) has been shown to elicit depressor responses [D. J. McKitrick and F. R. Calaresu. Am. J. Physiol. 255 (Regulatory Integrative Comp. Physiol. 24): R182-R187, 1988], we investigated the possibility that these responses might be facilitated either by electrical stimulation of arterial baroreceptor fibers in the aortic depressor nerve (ADN) or by simultaneous microinjection of L-glutamate (Glu) into the same sites in the NTS. Male Wistar rats (n = 51) were anesthetized with urethan (1.4 g/kg ip), artificially ventilated, and the dorsal medulla was exposed. The ADN was isolated, cut distally, and the central end was placed on bipolar stimulating electrodes. Threshold doses of 10(-7) M ANF microinjected into the NTS were combined with threshold electrical stimulation of the ADN (n = 37) or threshold doses of 0.13 0.5 M Glu (n = 14) microinjected into the NTS. There was a significant interaction between ANF microinjection and ADN stimulation in producing changes in mean arterial pressure (MAP) and heart rate [HR; P less than 0.05; -20.2 +/- 2.3 (SE) mmHg and -30.8 +/- 6.9 (SE) beats/min, respectively; n = 18]. There was also a significant interaction between ANF and Glu in producing changes in MAP and HR [P less than 0.05; -16.3 +/- 1.8 (SE) mmHg and -15.0 +/- 3.0 (SE) beats/min, respectively; n = 8]. These results indicate that ANF influences neurons in the NTS, which are also influenced by activation of arterial baroreceptors, and ANF and Glu interact in the NTS to produce facilitated cardiovascular responses. PMID- 2569841 TI - Use of "addictive" to describe neuroleptic drugs. PMID- 2569842 TI - Clinical differentiation between lethal catatonia and neuroleptic malignant syndrome. PMID- 2569843 TI - Benzodiazepine therapy and withdrawal in elderly patients. PMID- 2569844 TI - Hodgkins and large-cell lymphoma: a response to Dr. Whittaker. PMID- 2569845 TI - Epidemic hemorrhagic fever with renal syndrome in Yugoslavia, 1986. AB - An epidemic of hemorrhagic fever with renal syndrome (HFRS) occurred in Yugoslavia May-November 1986; all Republics and Provinces were involved. Serum samples were received from 260 of 276 persons with symptoms clinically compatible with a diagnosis of HFRS. Presumptive infection with a hantavirus was determined serologically for 161 of these. Many patients with serious clinical pictures, including severe renal insufficiency and shock, were hospitalized; 11 died. Indirect fluorescent antibody tests with antigens of 4 hantaviruses (Hantaan, Fojnica, Puumala, and the Vranica strain of Puumala virus) showed that greater than 1 serotype was circulating during this epidemic. Hantavirus antigens were detected in the lungs of 86 of 302 (28.5%) wild-caught small mammals. PMID- 2569846 TI - Hemorrhagic fever with renal syndrome in Yugoslavia: antigenic characterization of hantaviruses isolated from Apodemus flavicollis and Clethrionomys glareolus. AB - Hantavirus antigens were detected in lung tissues of 8/113 Apodemus flavicollis and 2/17 Clethrionomys glareolus captured in 1984 in Fojnica, a region of Yugoslavia endemic for hemorrhagic fever with renal syndrome; hantavirus antigens were not detected in lung tissues from 126 other mammals collected in Fojnica. Three hantaviruses, 2 from A. flavicollis and 1 from C. glareolus, were isolated directly in Vero E6 cells and were partially characterized. The isolates from A. flavicollis, designated Fojnica virus, were antigenically similar but not identical to Hantaan virus strain 76-118, whereas the isolate from C. glareolus was antigenically indistinguishable from Puumala virus, strain Hallnas B1. These data are consistent with previous studies that indicate the existence of at least 2 hantavirus serotypes in Yugoslavia. PMID- 2569847 TI - [The regulation of prolactin secretion]. PMID- 2569848 TI - [Vecuronium bromide and succinylcholine procedures in medial relaxation. A comparison of electromyography and clinical findings]. AB - Clinical and electromyographic effects of either succinylcholine (Suc) or vecuronium bromide (VEC) were compared during induction and maintenance of neuromuscular blockade for pelvic laparoscopy. METHODS: Forty ASA class I and II patients (pat.) were studied under general anesthesia with thiopental, enflurane, and nitrous oxide. Group VEC-pat. (n = 20) received 0.015 mg/kg body wt. VEC as priming and 5 min later 0.085 mg/kg as intubation doses. Repetitive doses of 0.01 mg/kg were injected to maintain twitch depression (T1%) less than or equal to 15%. Neuromuscular block was reversed with atropine and pyridostigmine (0.01 resp. 0.1 mg/kg). In group Suc-pat. relaxation was induced with 1.5 mg/kg Suc 5 min after pretreatment with 2 mg alcuronium. Relaxation (10% less than or equal to T1% less than or equal to 15%) was prolonged using a Suc infusion. Neuromuscular blockade was assessed electromyographically (Relaxograph, Datex) using train-of-four (TOF) stimulation (2 Hz for 2 s). Intubation conditions were scored according to Fahey et al. RESULTS: Pretreatment with 0.015 mg/kg VEC compared to 2 mg alcuronium led to a more pronounced decline in T1% and TOF-ratio (P less than 0.001). The time interval between injection of the intubation dose and complete relaxation (T1% less than or equal to 5%) was shorter in group Suc- than in VEC-pat. (P less than 0.001). Suc provides better intubation conditions than VEC (P less than 0.05). Recovery from muscle relaxation was faster in Suc- than in VEC-pat. (P less than 0.001). During Suc infusion in 8 patients a phase II block (TOF ratio less than or equal to 30%) was observed. DISCUSSION: Postoperative problems are often related to an unrecognized after effects of relaxants. Suc infusion leads to a remarkable number of phase-II blocks, whereas VEC can be antagonized promptly. PMID- 2569849 TI - Esmolol for potentiation of nitroprusside-induced hypotension: impact on the cardiovascular, adrenergic, and renin-angiotensin systems in man. AB - Esmolol infusion at rates of 200, 300, and 400 micrograms.kg-1.min-1 was used to potentiate hypotension (mean arterial pressure = 60 mm Hg) induced with sodium nitroprusside (SNP) in 10 male patients undergoing radical cancer surgery during nitrous oxide-oxygen and fentanyl anesthesia. Heart rate (HR), blood pressure (radial arterial catheter), and plasma levels of renin activity (PRA), norepinephrine (N), epinephrine (E), and dopamine (D) were measured: 1) while patients were awake; 2) after induction of anesthesia (nitrous oxide, 60% in oxygen, fentanyl = 5 micrograms/kg followed by an infusion at 10 micrograms.kg 1.hr-1); 3) after surgery had begun; 4) after 20 minutes of SNP-induced hypotension; 5) after 20 minutes of esmolol at each of the above infusion rates; and 6) after the completion of surgery. Compared to awake values, SNP-induced hypotension (mean infusion rate = 3.1 micrograms.kg-1.min-1 +/- 0.6 SE) during surgery resulted in significant (P less than 0.05) increases in heart rate, PRA, N, and D. Infusion of esmolol resulted in significant (P less than 0.05) dose dependent reductions in SNP requirement to maintain MAP = 60 mm Hg. At 200 micrograms.kg-1.min-1, SNP requirement was 2.1 micrograms.kg-1.min-1 +/- 0.4, at 300 micrograms.kg-1.min-1, it was 1.0 micrograms.kg-1.min-1 +/- 0.2, and at 400 micrograms.kg-1.min-1, was 0.5 micrograms.kg-1.min-1 +/- 0.3. Concomitant with the decrease in SNP requirement, there were significant reductions in HR and PRA at all infusion rates of esmolol.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569850 TI - Human IgE and IgG antibodies to mosquito proteins detected by the immunoblot technique. AB - Immunoblot technique was used in this study to detect IgE and IgG antibodies in human sera against mosquito antigens. Mosquito proteins were separated on SDS polyacrylamide gels and transferred electrophoretically to nitrocellulose papers. After incubation with sera from different individuals, the precipitated bands were analyzed with enzyme-labeled goat anti-human immunoglobulins. Distinct patterns of antigen were recognized by IgE and IgG antibodies. PMID- 2569851 TI - Efficacy of charcoal cathartic versus ipecac in reducing serum acetaminophen in a simulated overdose. AB - The traditional role of gastric emptying as the initial step in the management of the poisoned patient has recently been questioned; immediate activated charcoal administration has been recommended by some. In the setting of acetaminophen overdose, ipecac-induced emesis may interfere with subsequent oral antidotal therapy. Therefore, we conducted a study to compare the efficacy of initial therapy with ipecac with therapy with activated charcoal-cathartic in a simulated acetaminophen overdosage. Ten healthy volunteers participated in a randomized, crossover trial. Subjects ingested 3.0 g acetaminophen, followed by either no intervention, 30 mL syrup of ipecac, or 50 g activated charcoal-sorbitol solution at one hour. Serial acetaminophen levels were determined at intervals over eight hours. Both interventions significantly reduced the area under the curve compared with control (P less than .05). When comparing ipecac with activated charcoal cathartic, no significant difference was noted among these groups. PMID- 2569852 TI - Priapism: current concepts. AB - The treatment options for the management of the patient with priapism have changed markedly within the past several years. When possible, the underlying cause of the priapism should be identified. Therapy should be guided by the results of aspiration of the blood-filled corpora cavernosa of the erect penis. Early intervention through pharmacologic manipulation or surgical shunting should not be delayed while trying conservative measures. PMID- 2569853 TI - Influence of bovine intestinal fluid on the expression of K99 pili by Escherichia coli. AB - A study was conducted to determine whether intestinal fluid collected from various portions of bovine intestine differed in its effect on production of K99 pili by Escherichia coli. The small and large intestines of 7 calves, euthanatized 4 hours after a final feeding of milk, were divided into 6 to 9 segments from which intraluminal fluids were collected. Depending on the amount of fluid collected, up to 20 E coli strains that express K99 pili were grown on media prepared from the content of each specimen and then were tested for K99 pilus expression. In general, intestinal fluid from the most proximal small intestinal segments were more suppressive to K99 pilus expression than was fluid from more distal segments of small intestine. Only about 20% of the E coli test strains expressed K99 pili when grown on medium prepared from proximal small intestinal segmental fluid, whereas greater than 90% did when grown on medium prepared from distal small intestinal segmental fluid. Fluid from the large intestine varied considerably from calf to calf in its effect on K99 pilus expression. A correlation was found between K99 pilus expression and pH of the intestinal fluid, with the lower pH values (characteristic of proximal intestinal segmental fluid) being suppressive. The correlation between K99 pilus production and the pH of the medium was verified, using defined laboratory media adjusted to various pH values. Strains of E coli grown in medium at or below pH 5.5 failed to express K99 pili, whereas the same strains when grown in medium at or above pH 6.5 expressed K99 pili in abundance. PMID- 2569854 TI - DNA polymorphism analysis of hereditary multiple exostoses in horses. AB - Genomic DNA polymorphisms obtained by restriction fragment-length polymorphism from healthy horses and horses with hereditary multiple exostoses were analyzed. These DNA were digested by 12 restriction enzymes and were hybridized against 6 isotopically labeled oncogene probes. Hybridization was not detected with the viral oncogene, v-ras, which indicated this oncogene was absent in the equine genome. Oncogenes (c-raf-1, c-fes, c-myb, c-myc, and c-sis) were present and had similar hybridization patterns and signal intensities in DNA from healthy horses and horses with hereditary multiple exostoses. Unique and distinct restriction fragment-length polymorphisms were detected with the c-raf-1 probe only in BamHI- and PstI-digested equine DNA. PMID- 2569855 TI - Expression of the P-glycoprotein gene in multidrug-resistant Chinese hamster ovary cells. AB - A series of multidrug-resistant (MDR) Chinese hamster ovary (CHO) cells was established to survive in stepwise increasing concentrations of vincristine. These MDR cells contained amplified P-glycoprotein (mdr) genes. Using monoclonal antibody against the P-glycoprotein, we present here results showing that the levels of P-glycoprotein and its phosphorylation and glycosylation did not correlate with those of drug resistance. Our results suggest that overproduction of the P-glycoprotein cannot account for the differences in drug resistance in these MDR cells, and that multiple modalities are involved in multiple drug resistance in mammalian cells. PMID- 2569856 TI - PGE2 and PGA2 affect the allosteric properties and the activities of calmodulin dependent guanylate cyclase and Ca2+-stimulated ATPase of Walker-256 tumour microsomal membranes. AB - PGE2 and PGA2 incubated for 30 min at 25 degrees C with microsomal membranes isolated from Walker-256 tumour, in the presence of 50 microM indomethacin increase the lipid fluidity estimated by steady-state fluorescence anisotropy [(r0/r)-1]-1, using 1,6-diphenyl-1,3,5-hexatriene (DPH) as probe. The microsomal preparations of Walker-256 tumour contained calcium-stimulated and magnesium dependent ATPase as well as calmoduling-dependent guanylate cyclese activities. A considerable decrease (approx. 65%) in the activity of the Ca2+-stimulated ATPase was observed when preparations were treated with 10 microM PGE2 and PGA2. A dramatic gradual decrease of the calmodulin-dependent guanylate cyclase activity was also observed at different concentrations of PGE2 and PGA2 (0.25-10 microM). The ATP-dependent uptake of calcium was reduced by approximately 60% in microsomal membranes treated with PGE2 and PGA2. The allosteric properties of Ca2+-stimulated ATPase by Na+, and of guanylate cyclase by Mn.GTP (as reflected by changes in the Hill coefficients, h) were modulated by PGE2 and PGA2. The apparent cooperativity of the Ca2+-ATPase (h + 1.73 +/- 0.21) in control membranes was abolished (h + 1.1 +/- 0.11 and h = 0.9 +/- 0.09) in membranes treated by PGE2 and PGA2 (10 microM), while the allosteric stimulation of guanylate cyclase by Mn.GTP was reduced from h = 2.78 +/- 0.24 in control membranes to h = 1.92 +/- 0.16 and h = 1.73 +/- 0.15 in membranes treated by PGE2 and PGA2 (10 microM), respectively, suggesting that the physical state of Ca2+ stimulated ATPase and guanylate cyclase lipid microenvironments changed from a gel phase to a liquid-crystalline phase. In conclusion, it is suggested that PGE2 and PGA2 promote a phase separation in Walker-256 tumour microsomal membranes. This may be relevant to the Ca2+-calmodulin system and tumour growth inhibition. PMID- 2569857 TI - [Evaluation of the efficacy and safety of dilevalol in the treatment of essential arterial hypertension]. AB - Dilevalol, an optical isomer of labetalol, combines beta-blocking and vasodilator actions which prevent reflex tachycardia and it also has intrinsic beta-2-agonist action. Its low alpha-blocking action prevents orthostatic hypotension and improves first dose tolerance. The present study evaluates the efficacy and safety of dilevalol comparing it with captopril and nifedipine. Fifty-eight mild to moderate essential hypertensive patients were studied. After a 2 to 3 week placebo lead-in phase, patients with diastolic blood pressure between 95 and 105 mmHg were included., Patients were randomly assigned to one of the 3 study drugs for a 4-week period. Initially, 15 patients received dilevalol 200 mg qd and 15 captopril 100 mg bid; afterwards, 14 received dilevalol 200 mg qd and 14 nifedipine 40 mg bid. In the first group dilevalol decreased mean blood pressure (MBP) from 118.0 +/- 1.3 to 107.3 +/- 2.7 mmHg at week 4 and captopril from 117.8 +/- 1.2 to 108.6 +/- 2.7 (p less than 0.05). When dilevalol and nifedipine were compared, the former decreased MBP from 116.0 +/- 2.0 to 104.3 +/- 3.2 mmHg and the latter from 115.0 +/- 1.4 to 106.0 +/- 3.0 (p less than 0.05). The drug was well tolerated without evidence of orthostatic hypotension. Our results demonstrate that dilevalol is as safe and effective as captopril and nifedipine for the treatment of essential hypertension. PMID- 2569858 TI - Comparison of DNA restriction fragment length polymorphisms of Nostoc strains in and from cycads. AB - DNA was prepared from cyanobacteria freshly isolated from coralloid roots of natural populations of five cycad species: Ceratozamia mexicana mexicana (Mexico), C. mexicana robusta (Mexico), Dioon spinulosum (Mexico), Zamia furfuraceae (Mexico) and Z. skinneri (Costa Rica). Using the Southern blot technique and cloned Anabaena PCC 7120 nifK and glnA genes as probes, restriction fragment length polymorphisms of these cyanobacterial symbionts were compared. The five cyanobacterial preparations showed differences in the sizes of their DNA fragments hybridizing with both probes, indicating that different cyanobacterial species and/or strains were in the symbiotic associations. On the other hand, a similar comparison of cyanobacteria freshly collected from a single Encephalartos altensteinii coralloid root and from three independently subcultured isolates from the same coralloid root revealed that these were likely to be one and the same organism. Moreover, the complexity of restriction patterns shows that a mixture of Nostoc strains can associate with a single cycad species although a single cyanobacterial strain can predominate in the root of a single cycad plant. Thus, a wide range of Nostoc strains appear to associate with the coralloid roots of cycads. PMID- 2569859 TI - [The antagonism of ketamine/xylazine anesthesia ("Hellabrunn mixture") in wild zoo ruminants]. AB - The ability of the antagonists tolazoline, yohimbine and the combination of yohimbine with 4-aminopyridine to reverse the effects of the xylazine-component of the "Hellabrunn mixture" (125 mg/ml xylazine and 100 mg/ml ketamine) on nondomestic zoo ruminants is discussed. Arousal time, recovery time and changes in the parameter of circulatory and respiratory functions after antagonization are shown. Tolazoline is able to antagonize the xylazine effect completely within a short time. Using a dosage of 3-5 mg/kg there is a marked negative effect on the cardio-vascular system. Yohimbine in the used dosage of 0.25-0.3 mg/kg in non domestic ruminants did not approve in its effects. Combining yohimbine (0.25-0.3 mg) with 4-aminopyridine (0.5 mg/kg) recovery time is about 30 minutes. The negative effect on the cardio-vascular system is less pronounced compared with tolazoline. PMID- 2569860 TI - Methylmalonyl-CoA mutase from Propionibacterium shermanii. Evidence for the presence of two masked cysteine residues. AB - Adenosylcobalamin-dependent methylmalonyl-CoA mutase from Propionibacterium shermanii contains no intramolecular disulphide bridges, but two of the six thiol groups in the heterodimer are only revealed after reduction of the denatured enzyme with dithiothreitol. The available evidence suggests that they are present in disulphide linkages to unknown thiols of low Mr. The two specifically masked cysteine residues are Cys-535 in the alpha-subunit and Cys-517 in the beta subunit, which occupy exactly homologous positions in each chain. PMID- 2569861 TI - Cloning and structural characterization of the genes coding for adenosylcobalamin dependent methylmalonyl-CoA mutase from Propionibacterium shermanii. AB - The structural genes coding for both subunits of adenosylcobalamin-dependent methylmalonyl-CoA mutase from the Gram-positive bacterium Propionibacterium shermanii have been cloned, with the use of synthetic oligonucleotides as primary hybridization probes. The genes are closely linked and are transcribed in the same direction. Nucleotide sequence analysis of 4.5 kb of DNA encompassing both genes allowed us to infer the complete amino acid sequence of the two subunits: the beta-subunit is the product of the upstream gene, and consists of 638 amino acid residues (Mr 69465) and the alpha-subunit consists of 728 amino acid residues (Mr 80,147). There is a very close structural homology between the two subunits, reflecting the probable duplication of a common ancestral gene. A sequence present only in the alpha-subunit is significantly homologous to a portion of the sequence of the methylmalonyl-CoA-binding subunit of transcarboxylase from P. shermanii [Samols, Thornton, Murtif, Kumar, Haase & Wood (1988) J. Biol. Chem. 263, 6461-6464], and this homologous region may form part of the CoA ester-binding site in both enzymes. PMID- 2569862 TI - Subunit interactions in Propionibacterium shermanii methylmalonyl-CoA mutase studied by analytical ultracentrifugation. AB - The effect of increasing ionic strength on adenosylcobalamin-dependent methylmalonyl-CoA mutase from Propionibacterium shermanii was studied by using analytical ultracentrifugation. Both sedimentation-velocity and low-speed sedimentation-equilibration measurements show that the enzyme dissociates progressively into its two dissimilar subunits with increasing ionic strength. Equilibrium between the alpha beta-dimer and the separated subunits is rapidly established under these conditions. Dissociation is accompanied by loss of enzymic activity, but the position of the equilibrium is unaffected by the presence of either substrate or adenosylcobalamin cofactor. PMID- 2569863 TI - Co-ordinate regulation of low-density-lipoprotein receptor and 3-hydroxy-3 methylglutaryl-CoA reductase and synthase gene expression in HepG2 cells. AB - Cellular processes responsible for maintaining cholesterol homoeostasis are highly regulated. To determine whether two of these processes, cholesterol biosynthesis and receptor-mediated uptake of low-density lipoprotein (LDL), are co-ordinately regulated in human liver, we employed a human hepatoma cell line (HepG2) and measured the accumulation of mRNA for LDL receptor, 3-hydroxy-3 methylglutaryl-CoA (HMG-CoA) reductase and HMG-CoA synthase under a variety of conditions. Genomic Southern-blot analysis demonstrated that the integrity of these genes is maintained in the transformed cell. Treatment of HepG2 cells with mevalonate, 25-hydroxycholesterol, LDL, lovastatin or miconazole resulted in a similar effect on the accumulation of all three mRNAs at the concentrations tested. The onset of the response to drug, whether repression or induction of mRNA accumulation, occurred after approximately the same period of exposure for each mRNA. We conclude that the expression of the LDL receptor, HMG-CoA reductase and HMG-CoA synthase is co-ordinately regulated in HepG2 cells. PMID- 2569864 TI - Mitochondrial acetyl-CoA carboxylase. Time course of mobilization/activation in liver of refed rats. AB - Fasted (48 h) rats were killed at 0, 2, 4, 6, 8, 12, 16, 20 and 24 h after they were refed on a high-carbohydrate diet. An increase in the maximal activity and quantity of cystolic acetyl-CoA carboxylase was found in liver of refed rats after a lag time of about 8 h. The increased quantity of cytosolic enzyme was attributable primarily to mobilization of mitochondrial storage forms and not to substantial increase in the rate of synthesis of acetyl-CoA carboxylase. PMID- 2569865 TI - Atrial natriuretic peptide receptors and activation of guanylate cyclase in rat cardiac sarcolemma. AB - Two classes of atrial natriuretic peptide (ANP) receptors are present in purified sarcolemmal membrane fractions isolated from rat ventricle. Scatchard analysis using [125I]-ANP reveals high affinity (Kd approximately 10(-11) M) and low affinity (Kd approximately 10(-9) M) binding sites. Basal guanylate cyclase activities associated with these membrane fractions range from 3.2 +/- 1.3 pmol/min/mg protein in the presence of Mg2+ to 129 +/- 17 pmol/min/mg protein in the presence of Mn2+. Millimolar concentrations of adenosine triphosphate (ATP) potentiates Mg2+- but not Mn2+-supported activity. Binding of ANP to the low affinity site but not the high affinity site results in a maximum 2-fold activation of Mn2+- and up to 6-fold activation of Mg2+/ATP supported guanylate cyclase activities. PMID- 2569866 TI - The role of endosomal traffic in the transendothelial transport of ceruloplasmin in the liver. AB - Through a process resembling receptor-mediated endocytosis, liver endothelium binds and internalizes the plasma glycoprotein ceruloplasmin (CP) on the luminal side. The protein is then transported via a vesicular system to the albuminal side where it is externalized to the space of Disse. In its path, the glycoprotein is fully desialylated. To determine if the endosomal compartment is involved in this transport, we used endosomal inhibitors NH4Cl, ethylamine as well as monensin to quantitatively measure the magnitude of radiolabeled CP transport across purified liver endothelial cells. All three reagents inhibited the transport of CP and its discharge by endothelium. The magnitude of inhibition was dose-related for all three reagents. We conclude that the endosomal compartment is involved in the transendothelial transport of CP across the liver endothelium. PMID- 2569867 TI - The possible role of 9(S)-hydroperoxyoctadecatrienoic acid as a suicide substrate of soybean lipoxygenase. AB - While incubation of soybean lipoxygenase with alpha-linolenic acid resulted in the gradual decrease of lipoxygenase activity, the incubation with linoleic acid had no change. The inactivation of soybean lipoxygenase during incubation with alpha-linolenic acid was markedly observed at pH 6.5, but not at pH 9.0. Among the lipoxygenation products of alpha-linolenic acid, only 9(S) hydroperoxyoctadecatrienoic acid caused the inactivation of lipoxygenase. 9(S) Hydroxyoctadecatrienoic acid, 13(S)-hydroperoxyoctadecatrienoic acid or 9,16 dihydroperoxy conjugated trienoic acid was without effect. Accordingly, it is suggested that the epoxide intermediate, one conversion product of 9(S) hydroperoxyoctadecatrienoic acid, might be involved in the direct inactivation of lipoxygenase. PMID- 2569868 TI - GTP hydrolysis by guinea pig liver transglutaminase. AB - Homogeneous guinea pig liver transglutaminase was purified from a commercially available enzyme preparation by affinity chromatography on GTP-agarose. The purified transglutaminase exhibited a single band of apparent Mr = 80,000 on sodium dodecyl sulfate polyacrylamide gel and Western blotting and had enzyme activity of both transglutaminase and GTPase. The guinea pig liver transglutaminase has an apparent Km value of 4.4 microM for GTPase activity. GTPase activity was inhibited by guanine nucleotides in order GTP-gamma-S greater than GDP, but not by GMP. These results demonstrate that purified guinea pig liver transglutaminase catalyzes GTP hydrolysis. PMID- 2569869 TI - Photoaffinity labeling of P-glycoprotein in multidrug resistant cells with photoactive analogs of colchicine. AB - Two photoactive radiolabeled analogs of colchicine, N-(p-azido[3,5 [3H]benzoyl)aminohexanoyldeacetylcolchicine ([3H]NABC]) and N-(p-azido-[3 125I]salicyl)aminohexanoyldeacetylcolchicine ([125I]NASC) were synthesized and used to identify colchicine-specific acceptor(s) in membrane vesicles from multidrug resistant (MDR) variant DC-3F/VCRd-5L Chinese hamster lung cells. Both [3H]NABC and [125I]NASC specifically photolabeled a prominent 150-180 kDa polypeptide in membrane vesicles from DC-3F/VCRd-5L cells. The photolabeled polypeptide was immunoprecipitated by monoclonal antibody C219 specific for the MDR-related P-glycoprotein (P-gp) indicating the identity of this protein with P gp. Colchicine at 1000 microM reduced [3H]NABC photolabeling of P-gp by 72%. Furthermore, 100 microM of colchicine, vincristine, vinblastine, doxorubicin and actinomycin D inhibited [125I]NASC photolabeling by 45, 88.8, 91.1, 61.5, and 51% respectively. However, methotrexate did not affect the [125I]NASC photolabeling of P-gp, indicating the multidrug specificity of the P-gp colchicine acceptor for drugs to which these cells are resistant. PMID- 2569870 TI - Ochratoxin A decreases the activity of phosphoenolpyruvate carboxykinase and its mRNA content in primary cultures of rat kidney proximal convoluted tubule cells. AB - The effect of ochratoxin A on the specific activities of phosphoenolpyruvate carboxykinase, gamma-glutamyl transpeptidase and phosphate-dependent glutaminase were investigated in primary cultures of rat kidney proximal convoluted tubule cells grown in a serum-free medium supplemented with growth factors. Addition of ochratoxin A (25-100 microM) to the medium caused reduction in the specific activities of phosphoenolpyruvate carboxykinase and gamma-glutamyl transpeptidase only. Addition of cAMP caused a four-fold increase in the concentration of phosphoenolpyruvate carboxykinase mRNA in tubule cells within 3 h. This cAMP mediated increase in phosphoenolpyruvate carboxykinase mRNA content was abolished when ochratoxin A was added simultaneously. PMID- 2569872 TI - Effect of histamine H1-receptor antagonist on the regulation of carbohydrate and lipid metabolism in rat liver. AB - In order to elucidate the role of histamine in the liver, we studied the effect of a histamine H1-receptor antagonist on the carbohydrate and lipid metabolism in the rat liver. The administration of the H1-receptor antagonist decreased significantly the contents of glycogen and malonyl-CoA in the liver. However, it did not affect the levels of serum glucose and free fatty acid. These results suggest that histamine may play a part in the regulation of metabolism of carbohydrates and lipids in the liver. PMID- 2569873 TI - Induction of asparagine synthetase during lymphocyte activation by phytohemagglutinin. AB - Asparagine synthetase was increased in cultured mouse spleen lymphocytes after stimulation by phytohemagglutinin. After a lag period of about 24h, the enzyme activity level rose sharply by 48h, reached its maximum at 72h, and decreased thereafter. The time course of the change in the enzyme activity was similar to that of the change in the rate of DNA synthesis. From the results that there was no increase of the activity of asparagine synthetase at the time induction of ornithine decarboxylase would occur (6h), it seems unlikely that asparagine synthesized in the cells contributes to the enhancement of ornithine decarboxylase during the activation of lymphocytes. The increase of asparagine synthetase activity was inhibited by cycloheximide and somewhat by actinomycin D, suggesting de novo enzyme synthesis during the stimulation. PMID- 2569871 TI - A stimulatory effect of somatostatin: enhancement of activin A-mediated FSH secretion in rat pituitary cells. AB - The interaction between somatostatin and activin A was studied in terms of FSH secretion in rat pituitary cells in primary culture. Incubation of pituitary cells with 1 nM activin A for 48 hrs resulted in an increase in FSH release into incubation medium. The effect of activin A was dependent on cell-density and the higher the density, the smaller the stimulatory action of activin A. Somatostatin, by itself, did not affect the FSH secretion. When 100 nM somatostatin was included together with activin A or the cells were pretreated with somatostatin for 2 hrs, the activin A-induced FSH secretion was enhanced. This potentiation effect of somatostatin was inversely dependent on the cell density. These results indicate that somatostatin enhances, rather than inhibits, the activin A action in pituitary cells. PMID- 2569874 TI - Increased expression of p150,95 and CR3 leukocyte adhesion molecules by mononuclear phagocytes in rheumatoid synovial membranes. Comparison with osteoarthritic and normal synovial membranes. AB - The expression of leukocyte adhesion molecules CR3 (CD11b) and p150,95 (CD11c) in synovial tissue was evaluated immunohistochemically. Although a significant proportion of synoviocytes in normal and osteoarthritic synovial membranes expressed the leukocyte common antigen and CR3, very few expressed the p150,95 molecule. In contrast, p150,95 was more evident in rheumatoid synovial membranes. Expression of this molecule was strongest in synovial membranes with a prominent macrophage infiltrate and accumulation of mononuclear phagocytes at the joint surface; p150,95 was also present on interdigitating cells in lymphoid collections. The patterns of expression suggest that these leukocyte adhesion molecules may be important in the diapedesis of mononuclear phagocytes into and through inflamed synovial membranes, as well as in cellular interactions within rheumatoid synovial membranes. PMID- 2569875 TI - Short-term memory constraints upon the generation of diagnosis hypotheses by novice clinicians. AB - The structure and function of memory affect medical problem solving. The purpose of this 1982 study was to test the assertion that the number of diagnoses actively entertained by novice clinicians during a simulated case would not be greater than seven, the average maximum storage capacity of short-term memory. A total of 24 junior physician's assistant students took mock medical histories and gave mock physical examinations using simulated patients. Each of the eight cases investigated by each group included a primary problem with various numbers of secondary problems. Three successive time periods, called interludes, were provided in which each student had three minutes to list all the hypotheses he or she was actively considering concerning the diagnosis of the patient's problem. Each interlude was scheduled at a progressively more complete point in the student's knowledge of the case. The results supported the assertion that was tested. PMID- 2569876 TI - New possibilities for anti-migraine drugs: prostanoid antagonists and progesterone-mimicking stabilizers of excitable cells. AB - Medications currently available for the prophylaxis and treatment of migraine provide only limited relief. The pathophysiology of migraine is still poorly understood but it is widely accepted that various 'triggers', including fluctuations in the concentrations of circulating ovarian steroid levels, may initiate alterations in the activity of the intracranial vasculature and its efferent and afferent innervation. Recent pharmacological studies utilizing human intracranial artery preparations have addressed two distinct therapeutic stratagems. First, aspirin-like analgesics, which inhibit prostanoid synthesis, are widely used to treat migraine headache. Recent findings suggest that the ability of mefenamic acid to antagonise certain prostanoid actions, in addition to inhibiting synthesis, enhances its effectiveness. Thus, development of selective antagonists of the intracranial vasoconstrictor and the hyperalgesic actions of prostanoids could provide effective and selective migraine remedies. Second, with regard to prophylaxis, particularly of menstrually-related migraine, drugs which mimic the vascular smooth muscle 'stabilizing' action of ovarian steroids, possibly by enhancing potassium channel activation, are likely to be effective if used at concentrations that have minimal hypotensive effects. PMID- 2569877 TI - [Evaluation of astemizole in allergic conjunctivitis]. AB - A 4 week open study was carried out to evaluate the efficacy and safety of astemizole in the treatment of allergic conjunctivitis 31 patients were included in the study. Global efficacy, with 10 mg astemizole OD, was rated good or excellent in 100% of treated patients. A marked improvement of the main symptoms was noted at day 15. No side effects whether of the anticholinergic type or of the drowsiness type, were observed during the whole study. Astemizole appears to be a particularly interesting symptomatic treatment of allergic conjunctivitis. PMID- 2569878 TI - Interleukin-1 alpha gene intron containing variable repeat region coding for the SP1 transcription factor recognition sequence is polymorphic. AB - Interleukin-1 alpha (IL-1 alpha) is a cytokine produced by a number of cell types including macrophages, fibroblasts, keratinocytes, and mesangial cells. We were interested in identifying a DNA restriction fragment length polymorphism (RFLP) for the IL-1 alpha gene for use in studies of genetic alteration in various human cancers. Human genomic DNA from 32 unrelated individuals was digested with various restriction enzymes, alone and in combination, and subjected to Southern blot analysis. Hybridization to 32P-labeled IL-1 alpha cDNA revealed an insertion deletion-type polymorphic pattern. After digestion with RsaI, insertion-deletion type polymorphic bands with sizes of 3.4 kb, 3.1 kb, and 2.8 kb and one invariant band of 0.8 kb were observed. These three alleles, designated A1, A2, and A3, had relative frequencies of 0.18, 0.06, and 0.78 with heterozygosity observed in 38% of the unrelated individuals studied. Evaluation of nine related individuals for this RsaI polymorphism was consistent with a Mendelian inheritance. Comparison of restriction patterns following Southern analysis of DNA digested with several different enzymes showed that the polymorphic region resides within the sixth intron. Furthermore, this RFLP results from a variable length region containing multiple copies of a recognition sequence for SP1, an imperfect copy of viral enhancer elements, and an inverse and complementary sequence of the glucocorticoid receptor binding site. The identified polymorphism may be of value in analyses of chromosome 2 and may help to elucidate mechanisms by which IL-1 alpha transcription is regulated. PMID- 2569879 TI - [Studies on the relation of alkaloids in semen Strychni and its processing]. AB - This paper reports the determination of strychnine and brucine in different parts of Semen Strychni (the seeds of Strychnos nux-vomica and S. pierriana), and makes a comparison of the contents between the crude forms and processed products of the two seeds. Based on the results, the authors have made some discussion about the relationship between the alkaloid distribution in Semen Strychni and its processing. PMID- 2569880 TI - Immunohistology of the skin rash associated with acute HIV infection. AB - An immunohistological study of the rash associated with acute HIV infection in a homosexual man was undertaken. The salient feature was an infiltrate of mononuclear cells predominantly of the CD4+ phenotype around the superficial dermal vessels. HIV p24 antigen was detected in occasional cells, possibly Langerhans' cells, of the infiltrate. Their presentation of HIV antigen to local CD4+ cells may lead to a delayed type hypersensitivity reaction. PMID- 2569881 TI - [Modulation of the expression of histocompatibility antigens class I (HLA-A,B,C) in the human pancreas]. AB - In this study the expression of HLA class I molecules was analysed on caudal portions of ten pancreata from cadaver donors by means of indirect immunoperoxidase and immunophosphatase techniques. In 7 out of ten pancreata the results showed that islets tissue was almost negative for the expression of HLA Class I antigens as opposed to exocrine tissue that appeared positive. Within exocrine tissue and large sized islets strongly positive interstitial cells were also detected. Double stainings showed that the strongly positive interstitial cells expressed also Leu M1 antigens. Preliminary studies on the remaining three pancreata demonstrate an increase of hematic interstitial cells together with a parallel increase of HLA class I antigens by endocrine parenchyma. The above data suggest that an increase of interstitial cells within pancreas may influence islets antigenicity. PMID- 2569882 TI - Relationships among the cerebral amyloid peptides and their precursors. AB - Alzheimer's disease is characterized by deposits of amyloid in cerebral blood vessels and neuropil. Qualitative analyses of partially purified preparations of these amyloid deposits revealed the presence of a unique polypeptide now often called "beta peptide". This peptide is 40 residues long and it exhibits some amino terminal heterogeneity, which may result from the isolation procedure. The major amyloid peptide comprises at least 30% of the dry mass and 70% of the protein of washed neuritic plaque cores. These results indicate that the major peptide is the predominant proteinaceous component of cores; furthermore, they demonstrate that although cores may contain other substances such as aluminum silicate, polysaccharides, and lipids, amyloid peptide is a major component. More careful analysis reveals that the core amyloid peptide differs significantly from cerebrovascular amyloid peptide. Although the core amyloid peptide is constructed of the same backbone as the cerebrovascular amyloid peptide, it contains modifications that render the amino terminal region uncleavable by Edman degradation or by trypsin. It is unknown whether the lower solubility of core amyloid is related to these modifications. The original impetus for characterizing the differences between the core and cerebrovascular amyloid peptides arose from the question of whether both amyloid peptides were formed by a sequential pathway. Our results showing that core amyloid peptide is more extensively modified than vascular amyloid leads us to conclude that if a sequential pathway exists, vascular amyloid peptide must precede core amyloid peptide. Nevertheless, the discovery that amyloid precursor mRNA is widely and abundantly distributed throughout most tissues tends to discourage such a simple account of the relationship between these forms of amyloid.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569883 TI - The pathology of the amyloid A4 precursor of Alzheimer's disease. AB - The A4 amyloid protein is the major subunit present in the amyloid of Alzheimer's disease. It is derived by proteolytic cleavage from a larger precursor (PreA4) which is a neuronal membrane glycoprotein. Whereas in Down's syndrome, over expression of the gene coding for PreA4 is likely to be responsible for the premature development of cerebral amyloidosis, a similar mechanism is yet to be demonstrated in Alzheimer's disease. PMID- 2569884 TI - Betaxolol. AB - Drugs classified as beta-blockers have proved to be valuable in the treatment of patients with glaucoma. Timolol has become the most widely used ocular hypotensive agent. Actual and potential side effects associated with its non selective beta-blockade have prevented its use in patients with reactive airways disease, and to a lesser extent, with various cardiovascular conditions. Betaxolol is a relatively selective beta-1 blocker which in most patients is almost as effective as timolol in lowering intraocular pressure, and may be partly additive with dipivefrin. It is probably safer in patients unable to tolerate non-selective beta-blockers. However, it needs to be used with caution in these patients, who are unpredictably susceptible to systemic side effects. PMID- 2569885 TI - [Spinal subdural abscess--report of a case and a review of the literature of 43 cases]. AB - Spinal subdural abscess is rare, compared with spinal epidural abscess. The first case was reported by Sittig in 1927. Only 43 cases have been reported throughout the literature. We experienced an atypical fulminant case of spinal subdural abscess, following the operation for intracranial aneurysms. The patient was a 48 year-old male, admitted to our hospital on April 9, 1987 with complaining of sudden headache and vomiting. A lumbar puncture proved subarachnoid hemorrhage (S.A.H.), CT scan revealed saccular aneurysms in left middle cerebral artery and left internal carotid-ophthalmic artery. Radical surgical treatment for those aneurysms was undergone successfully. Neither cerebral ventricular nor cisternal drainage was established. His postoperative course was uneventful until the 9th postoperative day. He suddenly complained of pain in his legs and back, followed by paraplegia, respiratory, and cardiac arrests with clinical course of several hours. Antibiotics had been used for eight days after surgery, and there was no clinical evidence of inflammation sign. For his abrupt aggravation, a spinal infarction was suspected. However, the autopsy demonstrated that the dorsal side of the entire spinal cord was covered with masses of pus, and central necrosis and hemorrhage of the spinal cord mainly occupied the central gray matter; so called Pencilsharped softening. In this case, the cause of spinal subdural abscess was unclear. But the lumbar puncture performed on the day of admission, was most responsible. Also, the reasons why the clinical course of our case was rapid and fulminant were discussed with preference of spinal abscess to primary spinal circular disorder. 44 cases in the literature, including the presented report, was reviewed. Those symptomatology, predisposing factors, pathogenesis, the extent of spinal subdural abscess, clinical diagnosis, treatment, and prognosis were discussed. Among previous cases, this case was one of the most extensive subdural abscess and fulminant clinical course. PMID- 2569886 TI - Sufentanil disposition in patients undergoing renal transplantation: influence of choice of kinetic model. AB - We studied the disposition of sufentanil in 10 patients undergoing renal transplantation, and compared the data with those from eight healthy anaesthetized patients undergoing lower abdominal surgery. Patients received sufentanil 2.5 micrograms kg-1 as part of a balanced anaesthetic technique. Central venous blood samples were collected at intervals up to 600 min, and plasma sufentanil concentrations assayed by radioimmunoassay. Pharmacokinetic parameters were calculated from drug concentration-time profiles by extended least squares non-linear regression analysis (ELSFIT) and by a model independent (MI) approach using AUC and its first moment, AUMC. There were no differences (based on MI results) between the two groups for elimination half-life (T1/2 beta) (renal failure: 188 min; anaesthetized controls: 195 min), clearance (CI) (1030 and 1093 ml min-1) and apparent volume of distribution at steady state (Vss) (223.0 and 215.3 litre). Sufentanil binding to plasma proteins was 91.4% in the renal patients and 92.2% in the healthy group (ns). Comparison of kinetic methods showed significant correlation of the individual estimates for T1/2 beta, CI and Vss (P less than 0.01). The mean absolute differences between methods were: T1/2 beta 2.7 min (95% limits: -26.2 to 31.5), CI 36.5 ml min-1 (-5.5 to 78.4), Vss -18.4 litre (-47.7 to 10.9). When the mean estimate for the two methods [ELSFIT + MI)/2) was compared with the difference, there was no correlation for the estimates of CI and Vss+. MI tended to overestimate clearance and underestimate volume of distribution. There was a significant relationship between estimates for elimination half-life, with a slope greater than zero. PMID- 2569887 TI - Effects of temazepam premedication on cognitive recovery following alfentanil propofol anaesthesia. AB - The effects of temazepam 20 mg and placebo were compared for premedication in patients anaesthetized with propofol and alfentanil and undergoing day surgery. Temazepam 20 mg significantly reduced preoperative anxiety and increased recovery time. A series of computerized cognitive tasks revealed significant deficits in attention and memory following anaesthesia, which were increased in range and magnitude by temazepam, which were apparent 30 min after surgery and had largely, but not completely, recovered at 4 h. This study has demonstrated that computerized cognitive testing can identify a wider profile of impairments produced by temazepam than has been found in previous work using non-computerized techniques. PMID- 2569888 TI - Characterization of human neutrophil glycoproteins expressing the CD15 differentiation antigen (3-fucosyl-N-acetyllactosamine). AB - The expression of the CD15 antigen, 3-fucosyl N-acetyllactosamine, on neutrophil glycoproteins has been studied by SDS gel electrophoresis and immunoblotting. The antigen is expressed on several glycoproteins, both intracellularly and on the cell surface. Each subcellular compartment appears to contain a specific antigen. A soluble, granule glycoprotein (Mr 80-90K) probably accounts for most of the intracellular staining detected immunohistochemically. Membrane glycoproteins of Mr, 85-90K and 25K are associated with granule membranes, the latter being an integral membrane protein. The CD15 antigen is expressed on several cell surface glycoproteins with Mr in the range of 165K and 105K. These antigens are also contained in an intracellular pool which is brought to the surface on activation of the cells with chemotactic peptides. The 165K and 105K antigens show identical electrophoretic mobility to two of the major glycoproteins detectable by PAS or protein staining of gels of detergent extracts of cell membranes. These glycoproteins include the complement receptor, CR3. The beta chain of CR3 (105K) and to a lesser extent the alpha chain (165K) express CD15; however, most of the CD15 antigen is associated with other glycoproteins of these molecular masses. PMID- 2569889 TI - Picosecond-resolved fluorescence spectra of D-amino-acid oxidase. A new fluorescent species of the coenzyme. AB - Protein dynamics of D-amino-acid oxidase in the picosecond region was investigated by measuring time-resolved fluorescence of the bound coenzyme, FAD. The observed nonexponential fluorescence decay curves were analyzed with four exponential decay functions. The fluorescence lifetimes at the best fit were 26.6 +/- 0.7 ps, 44.0 +/- 4.2 ps, 177 +/- 11 ps, and 2.28 +/- 0.21 ns at 20 degrees C and 25.2 +/- 3.0 ps, 50.3 +/- 8.7 ps, 228 +/- 27 ps, and 2.75 +/- 0.33 ns at 5 degrees C. Component fractions with the shortest lifetime, ca. 26 ps, were always negative and close to -1. The other fluorescent components of the lifetimes, ca. 47 ps, 200 ps, and 2.6 ns, with positive fractions were assigned to different forms of the enzyme including the dimer, the monomer, and free FAD dissociated from the enzyme. Measurements of the time-resolved fluorescence spectra revealed that the maximum wavelengths of the spectra shifted toward shorter wavelength by 65 nm at 20 degrees C and 36 nm at 5 degrees C within 100 ps after pulsed excitation. The remarkable blue shift was not observed in free FAD. The first spectra immediately after the excitation of the enzyme exhibited maximum wavelengths of 584 nm at 20 degrees C and 557 nm at 5 degrees C. The fluorescence spectra obtained at times later than 100 ps are in good agreement with the one obtained under steady-state excitation of D-amino-acid oxidase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569890 TI - 31P NMR visibility and characterization of rat liver mitochondrial matrix adenine nucleotides. AB - Compartmentation and NMR visibility of mitochondrial adenine nucleotides were quantitated in isolated rat liver mitochondria respiring on succinate and glutamate in vitro at 8 and 25 degrees C. Intra- and extramitochondrial nucleotides were discriminated by adding the chelator trans-1,2 diaminocyclohexane-N,N,N',N'-tetraacetic acid (CDTA). T1 values of about 0.2-0.3 s for magnesium-bound matrix nucleotides were determined. Adenine nucleotide T1 values were influenced by the ionic environment; only magnesium-free ATP T1's were affected by temperature. Intra- and extramitochondrial adenine nucleotide ratios were varied in ATP-loaded mitochondria with added ATP and phosphate using the mitochondrial inhibitors oligomycin and carboxyatractyloside, and adenine nucleotides were quantitated by using NMR and enzymatic analysis. There was good agreement between matrix ATP concentrations (magnesium-bound ATP) calculated by using NMR and standard biochemical techniques. Although matrix ADP could be detected by NMR, it was difficult to quantitate accurately by NMR. The data indicate that mitochondrial ATP is NMR-visible in isolated mitochondria in vitro. PMID- 2569891 TI - Kinetic characterization of the [3'-32P]coenzyme A/acetyl coenzyme A exchange catalyzed by a three-subunit form of the carbon monoxide dehydrogenase/acetyl-CoA synthase from Clostridium thermoaceticum. AB - The ability of acetyl coenzyme A synthesizing carbon monoxide dehydrogenase isolated from Clostridium thermoaceticum to catalyze the exchange of [3' 32P]coenzyme A with acetyl coenzyme A is studied. This exchange is found to have a rate exceeding that of the acetyl coenzyme A carbonyl exchange also catalyzed by CO dehydrogenase ([1-14C]acetyl coenzyme A + CO in equilibrium acetyl coenzyme A + 14CO). These two exchanges are diagnostic of the ability of CO dehydrogenase to synthesize acetyl coenzyme A from a methyl group, coenzyme A, and carbon monoxide. The kinetic parameters for the coenzyme A exchange have been determined: Km(acetyl coenzyme A) = 1500 microM, Km(coenzyme A) = 50 microM, and Vmax = 2.5 mumol min-1 mg-1. Propionyl coenzyme A is shown to be a substrate (Km approximately 5 mM) for the coenzyme A exchange, with a rate 1/15 that of acetyl coenzyme A, but is not a substrate for the carbonyl exchange. CO dehydrogenase capable of catalyzing both these two exchanges, and the oxidation of CO to CO2, is isolated as a complex of molecular weight 410,000 consisting of three proteins in an alpha 2 beta 2 gamma 2 stoichiometry. The proposed gamma subunit, not previously reported as part of CO dehydrogenase, copurifies with the enzyme and has the same molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as the disulfide reductase previously separated from CO dehydrogenase in a final chromatographic step. PMID- 2569893 TI - Preparation of misacylated aminoacyl-tRNA(Phe)'s useful as probes of the ribosomal acceptor site. AB - Several pyroglutamylaminoacyl-tRNA's were prepared by T4 RNA ligase mediated condensation of synthetic pyroglutamylaminoacyl-pCpA's with tRNA's from which the last two nucleotides at the 3'-end had been removed. The derived pyroglutamylaminoacyl-tRNA's were incubated in the presence of calf liver pyroglutamate aminopeptidase, which effected their conversion to free aminoacyl tRNA's. The lack of contaminating esterase activities in the pyroglutamate aminopeptidase was verified by direct assay for the presence of the aminoacyl moieties in the formed aminoacyl-tRNA's and by the use of the deblocked aminoacyl tRNA's as acceptors in the peptidyltransferase reaction using an Escherichia coli ribosomal system. These findings provide the wherewithal for a detailed investigation of the substrate specificity of the peptidyltransferase center and for the elaboration of polypeptides containing modified amino acids at predetermined sites. PMID- 2569892 TI - Glutamic acid-88 is close to SH-1 in the tertiary structure of myosin subfragment 1. AB - The thiol-specific photoactivatable reagent benzophenone iodoacetamide (BPIA) can be selectively incorporated into the most reactive thiol, SH-1, of myosin S1, and upon photolysis, an intramolecular cross-link is formed between SH-1 and the N terminal 25-kDa region of S1. If a Mg2+-nucleotide is present during photolysis, cross-links can be formed either with the 25-kDa region or with the central 50 kDa region [Lu et al. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 6392]. Comparison of the peptide maps of cross-linked and un-cross-linked S1 heavy chains indicates that the segment located about 12-16 kDa from the N-terminus of the heavy chain can be cross-linked to SH-1 via BPIA independently of the presence of a nucleotide whereas the segment located 57-60 kDa from the N-terminus can be cross linked to SH-1 only in the presence of a Mg2+-nucleotide [Sutoh & Lu (1987) Biochemistry 26, 4511]. In this report, S1 was labeled with radioactive BPIA, photolyzed in the absence of nucleotide, and then degraded with proteolytic enzymes. Peptides containing cross-links were isolated by liquid chromatography and subjected to amino acid sequence analyses. The results show that Glu-88 is the major site and Asp-89 and Met-92 are the minor sites involved in cross linking with SH-1 (Cys-707) via BPIA. These residues are very near the reactive lysine residue (Lys-83) but relatively remote in the primary structure from the putative nucleotide binding region. PMID- 2569894 TI - Signal transduction by membrane receptors in viable electropermeabilized cells: isoproterenol-stimulated cyclic AMP synthesis in C6 glioma cells. AB - The activity of beta-adrenergic receptors at the plasma membrane level was investigated in viable, electropermeabilized C6 glioma cells. Electric field pulses were applied directly to the plated cells without any previous proteinase treatment. The affinity for isoproterenol and the density of the beta-adrenergic receptors, as judged from the number of [3H]CGP-12177 binding sites, were not affected by the electropermeabilization whereas the isoproterenol-stimulated cAMP accumulation was transiently impaired. This decrease in activity is due to an electropermeabilization-induced GTP leak. Normal activity could be obtained either by treating the cells by the electric field in a GTP-containing buffer, or by spontaneous recovery of the cells after the resealing of the plasma membrane, with a delay depending on the temperature. The activity of the receptors was not affected by the structural organization of the membrane associated to its electropermeabilization. PMID- 2569895 TI - [A mathematical model of the system controlling mediator release from presynaptic endings]. AB - A closed system of the first order enzymic reactions is considered as the model, one component controlling transmitter release. Examples are presented to demonstrate usefulness of the model. PMID- 2569896 TI - Carboxymethylation of an active site glutamic acid residue of ribonuclease F1 iodoacetate. AB - Ribonuclease (RNase) F1 was inactivated by incubation with an excess amount of iodoacetate at pH 5.5, 37 degrees C according to pseudo first-order kinetics. It was protected to various degrees, from inactivation by nucleotides, among which guanosine 2'-phosphate was most effective. The pseudo first-order rate constant was proportional to the reagent concentration, indicating that the reaction in reality follows second-order kinetics. The second-order rate constant was determined to be 25 x 10(-4) M-1 s-1. The inactivation rate was maximal at pH 5.5 6.0. When iodo[2-14C]acetate was used as the reagent, the stoichiometry of incorporation was determined to be 1.1 mol carboxymethyl group per mol of RNase F1 and glutamic acid residue 58 was assigned as the site of modification. PMID- 2569897 TI - Effect of idazoxan on evening melatonin plasma concentrations in healthy volunteers. PMID- 2569898 TI - Somatostatin concentration and binding in the rat hypothalamus and striatum during severe insulin-induced hypoglycemia. AB - Hypoglycemia was induced by administration of insulin (40 I.U./kg) to 24 h fasted rats. Somatostatin-like immunoreactivity (SLI) and 125I-Tyr11-somatostatin binding were measured in the striatum and hypothalamus at the onset of hypoglycemic coma (5-10 min). No significant changes in SLI concentration were detected in either site although the total number of specific somatostatin receptors in the striatum membranes, but not in the hypothalamus, decreased in insulin-injected rats when compared with the control group. PMID- 2569899 TI - Isolation and storage of peripheral blood hematopoietic stem cells for autotransplantation into children with cancer. AB - Peripheral blood stem cells (PBSC) were collected for autotransplantation by a total of 46 continuous-flow leukaphereses in 17 children with various types of cancer in whom the stem-cell pool had been expanded by chemotherapy. As the cells collected by leukapheresis were contaminated with many visible cell clumps, platelets, and erythrocytes, they were separated from the platelet-rich plasma (PRP) by slow-speed centrifugation and fractionated on a discontinuous gradient of Percoll. All the hematopoietic progenitors (CFU-GM, CFU-GEMM) in the starting samples were recovered at the interface of 40% and 60% Percoll solutions largely free of other cellular components and with a substantial reduction in volume. The separation and freezing procedures could be completed within three hours after obtaining cells by leukapheresis. After their fractionation and storage, these PBSC were shown to be able to reconstitute normal hematopoiesis in ten children with poor prognosis leukemia or neuroblastoma for whom no HLA-compatible marrow donors were available and who had been subjected to marrow-ablative therapy. This separation procedure is simple, efficient, and readily available and can be used for children as a routine procedure for PBSC autotransplantation. PMID- 2569900 TI - High-dose therapy and autologous peripheral blood stem cell transplantation for patients with lymphoma. AB - Forty patients with refractory Hodgkin's disease (24 patients) or non-Hodgkin's lymphoma (16 patients) who were considered for high-dose therapy but not for autologous bone marrow transplantation (ABMT) due to BM metastases, previous pelvic irradiation, a history of marrow involvement by tumor or hypocellular marrow in conventional harvest sites received high-dose therapy and autologous peripheral blood (PB) hematopoietic stem cell transplantation. Disappearance of circulating neutrophils and development of RBC and platelet transfusion dependence was followed, in the evaluable patients, by reappearance of 0.5 x 10(9)/L circulating granulocytes and sufficient platelets to obviate the need for platelet transfusions at a median of 25 days after transplantation. Twenty-three patients experienced a clinical complete remission (CR). The projected 2-year event-free survival was 24% for all 40 patients and 49% for the non-Hodgkin's lymphoma patients. The projected 18-month event-free survival for the Hodgkin's disease patients was 15%. PB stem cell transplantation provided an opportunity to administer high-dose salvage therapy to patients with refractory lymphoma who otherwise were not candidates for such therapy. For some of those patients, the high-dose therapy produced prolonged survival, free of tumor progression. PMID- 2569902 TI - Morphology of testicular germ cell tumours in treated and untreated cryptorchidism. AB - The histology of 75 testicular germ cell tumours in 73 patients with treated or untreated cryptorchidism was investigated in a group of 503 patients with testicular germ cell tumour and evaluated according to the WHO classification. The proportion of pure seminoma was associated with the height of the testis, being 87% in abdominal. 78% in inguinal and 50% in normally positioned testes. In patients operated upon for cryptorchidism, the current site of the testis seemed to be a more important determinant of this proportion than the original site. The proportion of pure seminoma which developed in testes after successful orchiopexy was equal to that in normally-descended testes (50%) and lower (39%) if orchiopexy had been performed before the age of 16 years. Similarly, among non seminomas, a higher proportion of tumours containing teratoma tissue was found in cryptorchidism was successfully treated in childhood. It was concluded that a successful orchiopexy in childhood decreases especially the risk of seminoma. PMID- 2569901 TI - Clinical course of primary HIV infection: consequences for subsequent course of infection. AB - OBJECTIVE: To investigate the impact of the clinical course of the primary HIV infection on the subsequent course of the infection. DESIGN: Prospective documenting of seroconversion, follow up at six month intervals, and analysis of disease progression by life tables. PATIENTS: 86 Men in whom seroconversion occurred within 12 months. PRIMARY OUTCOME MEASURE: Progression of HIV infection, defined as CD4 lymphocyte count less than 0.5 X 10(9)/l, recurrence of HIV antigenaemia, or progression to Centers for Disease Control group IV. MAIN RESULTS: Median follow up was 670 (range 45-1506) days. An acute illness like glandular fever occurred in 46 (53%) subjects. Three year progression rates to Centers for Disease Control group IV was 78% at three years for those who had longlasting illnesses (duration greater than or equal to 14 days) during seroconversion as compared with 10% for those who were free of symptoms or had mild illness. All six patients who developed AIDS had had longlasting primary illnesses. Three year progression rates to a CD4 lymphocyte count less than 0.5 X 10(9)/l and to recurrence of HIV antigenaemia were significantly higher for those who had longlasting primary illnesses than those who had no symptoms or mild illness (75% v 42% and 55% v 14%, respectively). CONCLUSION: The course of primary infection may determine the subsequent course of the infection. PMID- 2569903 TI - Corticotropin-releasing factor stimulates the release of methionine-enkephalin and dynorphin from the neostriatum and globus pallidus of the rat: in vitro and in vivo studies. AB - This study examined the changes in the in vitro and in vivo release of methionine enkephalin (Met-enkephalin), and dynorphin from the rat neostriatum in response to corticotropin-releasing factor (CRF). The levels of each opioid peptide were measured in the same sample collected at each time interval by specific radioimmunoassay methods. The in vitro release experiments were conducted using neostriatal slices (250 microns) obtained from adult male Wistar rats whereas in the in vivo studies, the release of both Met-enkephalin and dynorphin were assessed in push-pull perfusates of the caudate nucleus (containing both Met enkephalin and dynorphin cell bodies/fibres) and the globus pallidus (containing Met-enkephalin and dynorphin terminals) of chloral hydrate-anaesthetised adult male Wistar rats. In the in vitro studies, CRF (10(-12), 10(-10) and 10(-8) M) (applied in pulses of 75 min) stimulated both Met-enkephalin and dynorphin release from the neostriatal slices in a dose-related manner; in the presence of the CRF receptor antagonist, alpha-helical CRF9-41 (10(-6) M) the release of both Met-enkephalin and dynorphin in response to CRF (10(-8) M) were completely blocked. Push-pull perfusion experiments conducted in both the caudate nucleus and the globus pallidus, also demonstrated a dose-related increase in the release of both Met-enkephalin and dynorphin in response to CRF (10(-12), 10(-10) and 10( 8) M) applied in 60-min pulses. In addition, in each of the two brain sites, the release of both Met-enkephalin and dynorphin in response to CRF (10(-8) M) was completely blocked by alpha-helical CRF9-41 (10(-6) M). Both the in vitro and in vivo studies thus demonstrate that CRF can exert potent effects on Met-enkephalin and dynorphin release within the neostriatum-pallidum of the rat and that such effects are mediated via receptors specific for CRF, probably located on both the cell bodies and terminals of these opioid-containing neurones. The data obtained in this study thus substantiate the view that CRF, in addition to its regulation of pituitary opioid peptides, can communicate to opioid-containing neurones within the central nervous system and that many of the effects elicited by CRF may be ascribed to the opioid peptide released by CRF.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2569905 TI - An immunochemical and biochemical study of catecholaminergic activity in dissociated hypothalamic cultures. AB - Dopamine (DA)-containing cells of the medial basal hypothalamus (MBH) were dissociated and maintained in culture for up to 9 days. Cultures were evaluated both biochemically and immunochemically for DA activity. DA biosynthesis was determined using incorporation of [3H]tyrosine and was analyzed by HPLC with electrochemical detection. Immunochemical studies were performed to identify tyrosine hydroxylase (TH)-positive and neuron-specific enolase (NSE)-positive cells. Morphometric analyses determined the cell size, density, process length and the percent of neurons which were catecholaminergic. TH-positive neurons ranged from 6 to 8% of the total neuronal population when examined over days 3-9 of culture and the length of TH-positive neurites was significantly greater than that of NSE-positive cells. There was incorporation of [3H]tyrosine into DA as evidenced by the presence of [3H]DA in both the media and tissue and the inhibition of synthesis with alpha-methyl-p-tyrosine. There was a greater amount of labeled DA in the media than in the tissue at every time point examined. On the other hand, biosynthesis of DA in fresh brain tissue revealed approximately equal levels of DA in the media and tissue. These studies indicate that DA continues to be synthesized in dissociated cultures of MBH as evidenced by both the biochemical and immunochemical analyses and that there appears to be some alteration in the ability of these neurons to store the newly synthesized amine. PMID- 2569904 TI - Uptake of gamma-aminobutyric acid and L-glutamic acid by synaptosomes from postmortem human cerebral cortex: multiple sites, sodium dependence and effect of tissue preparation. AB - The uptake of gamma-aminobutyric acid (GABA) and L-glutamic acid by synaptosomes prepared from frozen postmortem human brain was shown to be effected via distinct high and low affinity sites. At approximately 17 h postmortem delay, the kinetic parameters for GABA uptake were: high affinity site, Km 7.1 +/- 2.5 microM, Vmax 18.7 +/- 4.8 nmol.min-1 per 100 mg protein; low affinity site, Km 2 +/- 1 mM, Vmax 425 +/- 250 nmol.min-1 per 100 mg protein (means +/- S.E.M., n = 13). Kinetic parameters for L-glutamate uptake were: high affinity site, Km 7.5 +/- 1.0 microM, Vmax 85 +/- 8 nmol.min-1 per 100 mg protein; low affinity site, Km 1.8 +/- 1.2 mM. Vmax 780 +/- 175 nmol.min-1 per 100 mg protein (n = 11). A detailed kinetic analysis of high affinity GABA uptake was performed over a range of sodium ion concentrations. The results were consistent with a coupling ratio of one Na+ ion to one GABA molecule; a similar result was found with rat brain synaptosomes. However, rat and human synaptosomes differed in the degree to which the substrate affinity of the high affinity GABA uptake site varied with decreasing Na+ ion concentration. High affinity GABA uptake was markedly affected by the method used to freeze and divide the tissue, but did not vary greatly in different cortical regions. There was some decline of high affinity GABA uptake activity with postmortem delay, apparently due to a loss of sites rather than a change in site affinity. PMID- 2569906 TI - Spinally projecting noradrenergic neurons of the dorsolateral pontine tegmentum: a combined immunocytochemical and retrograde labeling study. AB - Two to three days following injection of horseradish peroxidase (HRP) into the spinal cords of 5 cats, the animals were sacrificed and perfused, and the brainstems removed and sectioned. The sections were then processed for HRP and, immunocytochemically, for tyrosine hydroxylase (TH). The dorsolateral pontine tegmentum was divided into the locus coeruleus, subcoeruleus and Kolliker-Fuse nucleus; the mean percentage of pontospinal neurons containing TH were found to be 85.5 +/- 2.5 (S.E.M.), 79.6 +/- 5.6 and 87.1 +/- 3.1, respectively. The cell diameters of locus coeruleus cells were also measured. PMID- 2569907 TI - A general pattern of CNS innervation of the sympathetic outflow demonstrated by transneuronal pseudorabies viral infections. AB - Pseudorabies virus (PRV) injections of various sympathetic ganglia and the adrenal gland were made in rats. These produced immunohistochemically detectable retrograde viral infections of ipsilateral sympathetic preganglionic neurons (SPNs) and transneuronal infections of the specific sets of second order neurons in the spinal cord and brain that innervate the infected SPNs. Five cell groups in the brain appear to regulate the entire sympathetic outflow: the paraventricular hypothalamic nucleus (PVH), A5 noradrenergic cell group, caudal raphe region, rostral ventrolateral medulla, and ventromedial medulla. In addition, local interneurons in laminae VII and X of the spinal cord are also involved. Other CNS areas also became transneuronally labeled after infections of certain sympathetic ganglia, most notably the superior cervical and stellate ganglia. These areas include the central gray matter and lateral hypothalamic area. The zona incerta was uniquely labeled after stellate ganglion infections. The cell body labeling was specific. This specificity was demonstrated in the PVH where the neurons of the parvocellular PVH that form the descending sympathetic pathway were labeled in a topographic fashion. Finally, we demonstrate that the retrograde transneuronal viral cell body labeling method can be used simultaneously with either neuropeptide transmitter or transmitter synthetic enzyme immunohistochemistry. PMID- 2569908 TI - Feline subthalamic nucleus neurons contain glutamate-like but not GABA-like or glycine-like immunoreactivity. AB - The identity of the neurotransmitter of subthalamic nucleus neurons has not been definitively established. GABA, glycine, and glutamate have all been hypothesized to be the neurotransmitter of these neurons. Immunohistochemistry with 3 well characterized antisera against glutamate, GABA, and glycine were used to study feline subthalamic nucleus neurons. These neurons were found to contain intense glutamate-like but not GABA- or glycine-like immunoreactivity. The surrounding neuropil contained glutamate-like and GABA-like but not glycine-like immunoreactivity. These results support the hypothesis that subthalamic nucleus neurons are glutamatergic. PMID- 2569909 TI - Action of clonidine on micturitional reflexes in decerebrate cats. AB - We explored the urodynamic action of clonidine (preferential alpha 2-agonist) and yohimbine (preferential alpha 2-blocker) in decerebrate cats. These animals retain catecholaminergic fibers from the complex of the locus coeruleus to the spinal cord and have synergic and faster micturition cycles than normal cats. Twenty-nine male cats were made decerebrate at intercollicular level under ether anesthesia. Voiding, vesical, urethral and blood pressures, and the EMG of the external sphincter-pelvic floor and leg muscles were studied. Preparations with the urethrovesical junction opened and closed were used. Control activity was characterized by two types of vesical pressure waves: (1) low frequency, high intensity, and (2) high frequency, low intensity. Type-2 preceded Type-1 waves. Clonidine caused: (A) reduction of Type-1 and enhancement of Type-2 Pves waves; (B) diminution of vesical capacity; (C) facilitation of external sphincter relaxation; (D) inhibition of skeletal muscle activity; (E) systemic hypotension and bradycardia. Yohimbine inhibited clonidine's actions. When injected alone, it inhibited vesical and increased sphincteric activities. It also produced systemic hypertension and enhanced crisis of autonomic dysreflexia. The overall effect of clonidine on urodynamics in acute decerebrate cats was to bring about an active, small (normotonic) bladder with urinary frequencies. We suggest that clonidine: (1) inhibited Type-1 waves by inhibiting cellular activity in the locus coeruleus. As a result, the vesico-vesical contraction long loop reflex was abolished; (2) facilitated Type-2 waves by inhibiting inhibitory interneurons in the sacral parasympathetic center. As a result, the vesicovesical contraction short loop reflex was facilitated. PMID- 2569910 TI - Distribution of alpha 2-adrenergic receptors in the brain of the Japanese quail as determined by quantitative autoradiography: implications for the control of sexually dimorphic reproductive processes. AB - With the use of [3H]p-aminoclonidine (PAC), alpha 2-adrenergic binding sites were mapped in the brain of the Japanese quail (Coturnix coturnix japonica). The sites were labeled with the use of in vitro quantitative autoradiography. Special attention was given to areas implicated in the control of sexually dimorphic reproductive processes including sexual behavior. Preliminary competition experiments found that [3H]PAC binding on tissue sections exhibited a pharmacology appropriate to the alpha 2 receptor. Binding sites were found to be heterogeneously distributed throughout the brain. Some of the highest levels of specific binding were found in several areas regulating reproductive function such as the preoptic area, the supraoptic nucleus, the infundibulum, and the medial mammillary nucleus of the infundibulum. [3H]PAC labeled precisely the morphologically dimorphic preoptic medial nucleus but no sexual dimorphism in density of receptor binding was identified. However, dimorphism in density of receptor binding was identified in two areas: the medial mammillary nucleus and the mesencephalic intercollicular nucleus. The former area appears to be involved in the regulation of gonadotrophin secretion and the latter area has been implicated in the control of vocal behavior. These neurochemical dimorphisms may contribute to the regulation of two sexually dimorphic reproductive processes, gonadotropin secretion and courtship vocalizations. PMID- 2569911 TI - Spatial relationship of the striatonigral and mesostriatal pathways: double-label immunocytochemistry for DARPP-32 and tyrosine hydroxylase. AB - Antibodies to tyrosine hydroxylase and DARPP-32 were used to examine the spatial arrangement between mesostriatal dopamine projections and the reciprocal pathway from DARPP-32-containing neurons in the basal forebrain. Use of a double-labeling immunocytochemical procedure demonstrated that the mesostriatal and striatonigral pathways run in close proximity throughout the rostral mesencephalon and basal forebrain. The majority of descending axons immunoreactive for DARPP-32 appear to originate in the striatum, including the nucleus accumbens, and run through the internal capsule to innervate the globus pallidus, entopeduncular nucleus, and all subdivisions of the substantia nigra. The ventral tegmental area is sparsely invested with DARPP-32-immunoreactive axons. At all levels, there are also fascicles of DARPP-32-containing fibers which run ventromedial to the internal capsule in the medial forebrain bundle, and which are coextensive with ascending axons of the mesencephalic dopamine the internal capsule in the medial forebrain bundle, and which are coextensive with ascending axons of the mesencephalic dopamine cell groups. Tyrosine hydroxylase-immunoreactive axons are coextensive with DARPP-32-immunoreactive axons in the internal capsule entopeduncular nucleus, and globus pallidus, as well as much of the remainder of the basal forebrain. Although the main source of descending DARPP-32 immunoreactive axons would appear to be the striatum, other possible sources are also discussed. PMID- 2569912 TI - Noradrenergic mechanisms in hippocampal kindling with rapidly recurring seizures. AB - Rats were subjected to hippocampal kindling with short interstimulus intervals (5 min), so-called rapid kindling. Severe depletion of forebrain noradrenaline (NA) with intraventricular 6-hydroxydopamine or blockade of alpha 2 adrenergic receptors with idazoxan markedly reduced the number of stimulations needed to induce the first severe limbic seizure and increased the total number of such seizures during 40 stimulations. Intracerebral microdialysis demonstrated a 3 fold increase of extracellular NA levels in response to the first kindling stimulations in both the stimulated and the non-stimulated hippocampus. The NA levels then gradually tapered off reaching baseline levels after 14 stimulations. We conclude that central noradrenergic neurons exert a powerful suppressant action, most probably mediated via an alpha 2 adrenergic receptor, on seizure development in rapid hippocampal kindling. PMID- 2569913 TI - Selective depression of GABA-mediated IPSPs by somatostatin in area CA1 of rabbit hippocampal slices. AB - In area CA1 of hippocampus, a subpopulation of gamma-aminobutyric acid (GABA) containing interneurons that make synaptic contacts on pyramidal cells also contains the neuropeptide, somatostatin. The effects of GABA and somatostatin on hippocampal pyramidal cells have been investigated separately, but it is not known whether an interaction occurs between these co-localized substances. We demonstrate that somatostatin has a potent inhibitory effect on GABA-mediated synaptic potentials which hyperpolarize pyramidal cells. This effect may be relevant to the well-documented epileptogenicity of the hippocampus, as well as the phenomenon of long-term potentiation, which is a well-studied example of synaptic plasticity. PMID- 2569914 TI - Cytoplasmic loop of beta-adrenergic receptors: synaptic and intracellular localization and relation to catecholaminergic neurons in the nuclei of the solitary tracts. AB - Pharmacological studies suggest that beta-adrenergic receptors (beta AR) in the medial nuclei of the solitary tracts (m-NTS) facilitate presynaptic release of catecholamines and also function at postsynaptic sites. We have localized the antigenic sites for a monoclonal antibody against a peptide corresponding to amino acids 226-239 of beta AR in the m-NTS of rat brain. By light microscopy, immunoperoxidase labeling for this antibody was detected in somata and proximal processes of many small cells that were distributed throughout the rostrocaudal extent of the m-NTS. Electron microscopy confirmed the cytoplasmic localization of beta AR in perikarya and proximal dendrites of neurons. Immunoreactivity occurred as discrete patches associated with cytoplasmic surfaces of plasma membrane and with irregularly-shaped saccules with clear lumen in the immediate vicinity. Select regions of nuclear envelopes, mitochondrial membranes, and rough endoplasmic reticulum were also immunoreactive along their cytoplasmic surfaces. In contrast, the Golgi apparatus was labeled, but infrequently. Immunoreactivity was also detected at numerous post- and occasional presynaptic membrane specializations of select axodendritic junctions. Dual labeling for the beta AR antibody by the immunoperoxidase method and for a rabbit antiserum against the catecholamine-synthesizing enzyme, tyrosine hydroxylase (TH), by the immunoautoradiographic method within the same sections, further established the precise cellular relations between beta AR and catecholaminergic neurons. Immunoreactivity for beta AR was detected in numerous perikarya and proximal dendrites that did not show detectable levels of TH. However, a few cells were dually labeled for both antigens, as seen by both light and electron microscopy. The TH-labeled terminals formed synapses at junctions both with and without beta AR-like immunoreactivity. These results from the single and dual labeling studies: (1) confirm biochemical predictions that amino acids 226-239 of beta AR protein reside intracellularly; (2) provide the first ultrastructural evidence for beta AR localization within both pre- and postsynaptic membrane specializations of a subset of catecholaminergic synapses; and (3) suggest select intracellular sites that may be involved with synthesis and/or internalization and degradation of the receptor protein. PMID- 2569915 TI - Immunohistochemical studies on the somatostatin- and growth hormone-releasing factor (GRF)-neurons in the hypothalamus of the novel dwarf rat; the spontaneous dwarf rat (dr). AB - Immunoreactivities of somatostatin and growth hormone-releasing factor (GRF) in the hypothalamus of spontaneous dwarf rats (SDRs, gene symbol; dr), which show isolated growth hormone (GH) deficiency, and normal rats were studied with an avidin-biotin complex (ABC) immunohistochemical method. Somatostatin and GRF immunoreactivities were observed in the median eminence and each afferent nucleus with no difference between these animals. The observation suggests that the etiology of the GH deficiency lies in GH cells themselves and is not hypothalamic in origin. This observation indicates that the SDR is a new model animal for type I dwarfism. PMID- 2569916 TI - Direct synaptic contacts of catecholamine fibers on neuropeptide Y-containing neurons of the rat cerebral cortex. AB - A direct synapse between catecholamine (CA) fibers and neuropeptide Y (NPY) containing neurons was demonstrated in rat cerebral cortex by using an immunohistochemical double-staining method under electron microscopy. PMID- 2569917 TI - Complex responses activated by ibotenate in postnatal rat hippocampal neurons. AB - The actions of the excitatory amino acid ibotenate were investigated in postnatal rat hippocampal neurons. In neurons voltage clamped at negative membrane potentials using low-chloride internal solutions, ibotenate responses consist of an inward cationic current and two outward currents. The inward current is inhibited by 2-amino-5-phosphonovalerate (APV), a selective N-methyl-D-aspartate (NMDA) antagonist. The two outward currents consist of a picrotoxin and bicuculline-sensitive chloride current and a slowly developing calcium activated potassium current. The bicuculline sensitive current appears to be the product of contamination of ibotenate samples with the gamma-aminobutyric acid (GABA) agonist muscimol and not the result of a direct action of ibotenate. PMID- 2569918 TI - Effects of insulin-induced hypoglycemia on somatostatin level and binding in rat cerebral cortex and hippocampus. AB - The effects of severe insulin-induced hypoglycemia on somatostatin level and specific binding in the cerebral cortex and hippocampus were examined using 125I Tyr11-somatostatin as a ligand. Severe insulin-induced hypoglycemia did not affect the level of somatostatin-like immunoreactivity in the brain areas studied. However, the number (but not the affinity) of specific somatostatin receptors was significantly decreased in membrane preparation from the hippocampus but not in the cerebral cortex at the onset of hypoglycemic coma (5 10 min). Administration of glucose at the onset of hypoglycemic coma brought about extensive recovery of hippocampal somatostatin receptor number. These results suggest that glucose modulates the somatostatin receptor in the rat hippocampus. The physiological significance of these findings remains to be clarified. PMID- 2569919 TI - TRH stimulation and L-glutamic acid inhibition of proximal gastric motor activity in the rat dorsal vagal complex. AB - The importance of the dorsal vagal complex (DVC) in the control of gastric motor activity has been previously established by electrical and chemical stimulation of this region. We have further evaluated excitatory and inhibitory influences on motor activity of the gastric corpus by microinjection of L-glutamic acid (GLU) and thyrotropin-releasing hormone (TRH) into the DVC. GLU and TRH were ejected by pressure (20-30 psi) in 1-10 nl vol. from multibarreled micropipettes and intraluminal pressure in the gastric corpus was measured using a manometric catheter placed into the stomach through the pylorus of urethane-chloralose anesthetized rats. Gastric motor activity was monitored while micropipettes were advanced from the surface of the dorsomedial medulla to a depth of 1 mm in 100 micron increments. Microinjections of GLU (1-10 pmol) at depths of 200-600 microns below the surface of the brainstem caused a decrease in tonic intraluminal pressure and amplitude of phasic contractions of the gastric corpus. Injection of TRH (1-10 pmol) at depths of 200-800 microns increased both tonic intraluminal pressure and amplitude of phasic contractions. The responses to GLU (10 pmol) and TRH (10 pmol) were abolished by hexamethonium and vagotomy; atropine abolished the effect of TRH and attenuated that of GLU. It is concluded that GLU evokes only vagally mediated inhibitory effects on tonic and phasic gastric motor activity when microinjected into the DVC. In contrast, injection of TRH at the same loci causes only vagal cholinergic increases in motor activity. Subpopulations of neurons in the DVC may, therefore, be activated by specific neurotransmitters having opposite effects on gastric motor activity. PMID- 2569920 TI - Hippocampal interneuron loss and plasticity in human temporal lobe epilepsy. AB - It has been hypothesized on the basis of animal models of epilepsy that abnormal neural activity in epilepsy may be related to reorganized neural circuits that facilitate epileptogenesis. Little evidence of this was available for human epilepsy. This paper provides the first evidence of such reorganization of a hippocampal seizure focus in human temporal lobe epilepsy (TLE). This reorganization involves the selective loss of somatostatin and neuropeptide Y immunoreactive interneurons, and axonal sprouting of other neuropeptide Y neurons and dynorphin-A immunoreactive granule cells. This set of changes is not exactly like those that are reported in animal models. PMID- 2569921 TI - [Comparison of the effect of BL 343 Ac with beta-adrenolytics on heart muscle preparations]. AB - The paper evaluates the effect of the newly synthesized substance BL 343 Ac- propyl-4[(2-hydroxy-3-isopropylamino)-propoxy]carbanilate hydrochloride--on mechanical parameters of the myocardium of isolated guinea pig atria. At high concentrations of the substance a negative inotropic and chronotropic effect and prolongation of the functional refractory period of the preparations were recorded. These effects were comparable to those of propranolol and metipranolol. Furthermore, a mild increase in the rate of the reserpinized atria after application of 10(-7) mol.l-1 BL 343 Ac suggests a possible intrinsic sympathetic activity of the substance. BL 343 Ac has an antiisoprenaline effect with a pA2 value comparable to that of propranolol and metipranolol. However the slope of the Schild plot was different from 1, indicating that the substance possesses also other than competitively beta-adrenolytic properties. PMID- 2569923 TI - Characterization of the muscarinic and serotoninergic receptors of the intestine of the rainbow trout (Salmo gairdneri). AB - The ability of carbachol and 5-hydroxytryptamine (5-HT) to contract isolated segments of rainbow trout intestine in a concentration-dependent manner indicates the presence of muscarinic and serotoninergic receptors in this tissue. The activity of these agonists appears to be directly on the smooth muscle, since ganglionic blockers and inhibitors of neurotransmission did not inhibit contractions. The carbachol-induced contractions were selectively inhibited by atropine and (+-)-3-quinuclidinyl xanthene-9-carboxylate hemioxalate hydrate, an M-2 muscarinic receptor antagonist. However, the inhibition was not competitive. McN-A-343, an M-1 muscarinic agonist had no effect on intrinsic tone. The 5-HT induced contractions were selectively inhibited by methysergide and the 5-HT2 receptor blockers, ketanserin and 1-(1-naphthyl)piperazine. Again, the inhibition by these agents was not competitive. 5-HT1 and 5-HT3 receptor antagonists did not inhibit contractions. The results thus suggest that the smooth muscle of the rainbow trout intestine contains M-2 muscarinic and 5-HT2 receptors. PMID- 2569922 TI - A set of Macintosh computer programs for the design and analysis of synthetic genes. AB - Computer programs that can be used for the design of synthetic genes and that are run on an Apple Macintosh computer are described. These programs determine nucleic acid sequences encoding amino acid sequences. They select DNA sequences based on codon usage as specified by the user, and determine the placement of base changes that can be used to create restriction enzyme sites without altering the amino acid sequence. A new algorithm for finding restriction sites by translating the restriction endonuclease target sequence in all three reading frames and then searching the given peptide or protein amino acid sequence with these short restriction enzyme peptide sequences is described. Examples are given for the creation of synthetic DNA sequences for the bovine prethrombin-2 and ribonuclease A genes. PMID- 2569924 TI - An immunologic profile of young adults with head and neck cancer. AB - Numerous reports have suggested, although never demonstrated, a suppressed immune defense mechanism as a contributing factor in the development of head and neck cancer in the young adult. Twenty-four previously untreated adults less than or equal to 40 years of age with squamous cell carcinoma were examined for lymphocyte function (natural killer cell activity and in vitro lymphocyte blastogenesis response to mitogens), total lymphocyte number and percentage of lymphocyte subsets, and humoral immune status (circulating IgA, IgG, and IgM). As compared with 33 healthy young adults, no significant immunologic deficit could be identified. On the contrary, the young adult cancer population had significantly increased lymphocyte numbers (P less than 0.05) and serum IgA, IgG, and IgM levels (P less than 0.001, respectively). These young cancer patients cannot be considered to be immunosuppressed. Alternative biologic mechanisms must be defined to account for the increasing incidence of head and neck cancer over the last decade among young adults in the United States. PMID- 2569925 TI - The relationship between somatostatin, epidermal growth factor, and steroid hormone receptors in breast cancer. AB - The somatostatin (SS) and the epidermal growth factor (EGF) receptor content have been established in 36 primary breast cancers by receptor autoradiography on adjacent tissue sections. Iodine 125 (125I)-EGF was used as radioligand for EGF receptor visualization whereas an iodinated SS-28 analogue or an octapeptide SS analogue were used to measure SS receptors. Six of 36 tumors contained SS receptors, whereas ten of the 36 tumors were shown to contain EGF receptors. None of the tumor samples containing SS receptors were simultaneously EGF receptor positive. In contrast, all SS receptor-positive tumors simultaneously contained steroid receptors. The positive correlation between SS receptors and steroid receptors as well as the negative correlation between SS receptors and EGF receptors therefore suggest that the small percentage of SS receptor-positive breast tumors are a group of differentiated breast tumors with a good prognosis. In these cases, combined hormonetherapy including SS analogs may be of potential interest. PMID- 2569926 TI - Effect of the antihistamine, methapyrilene, as an initiator of hepatocarcinogenesis in female rats. AB - The antihistamine methapyrilene was examined for its ability to initiate hepatocarcinogenesis in rats. Rats were first subjected to partial hepatectomy and then were intubated with one of four doses (30, 100, 200 or 300 mg/kg) of methapyrilene hydrochloride (or an equivalent amount of water for controls, or 10 mg diethylnitrosamine/kg for positive controls). Rats were then fed 0.05% phenobarbital in the diet for 3, 6 or 9 months. The number and volume of altered hepatic foci were quantified with the histochemical markers gamma-glutamyl transpeptidase, glucose-6-phosphatase, and ATPase. The number of foci induced was increased 2- to 4-fold by the highest dose of methapyrilene at all 3 time points, but the only statistically significant increase was produced by the 200 mg/kg dose after 3 months of promotion. This study shows that methapyrilene may act as a weak initiator. PMID- 2569927 TI - Induction and promotion of gamma-glutamyltranspeptidase-positive foci in the liver of female rats treated with ethinyl estradiol, clomiphene, tamoxifen and their associations. AB - The objective of the present study was to determine whether a short exposure (6 weeks) to high doses of ethinyl estradiol (EE) could not only promote but also initiate hepatocarcinogenesis, and whether two antiestrogens, clomiphene (C) and tamoxifen (T), could influence EE activity. 2-Acetylaminofluorene (AAF), which has been shown to produce rat liver hyperplastic lesions characterized by the presence of estrogen receptors, was used either as a promoter to test for initiating activity, or as an initiator to test for promoting activity. Putative preneoplastic lesions were identified by means of a positive gamma glutamyltranspeptidase (GGT) reaction. The results revealed that when administered alone in female Sprague-Dawley rats, not only E, but also C and T were clearly active in both initiating and promoting the development of GGT positive foci. Moreover, in rats of the same strain treated with EE + C or EE + T a significant increase in the incidence of GGT foci demonstrated the occurrence of an additive effect in terms of both initiating and promoting activity. Fischer 344 rats were more susceptible than Sprague-Dawley rats to promotion by EE, C and T, but any substantial evidence of an additive effect was absent when the two anti-estrogens were administered in association with the estrogen. PMID- 2569928 TI - Differential expression of the c-erbB-2 gene in human small cell and non-small cell lung cancer. AB - We studied non-small cell lung cancers (NSCLC) from 60 patients for abnormalities in the c-erbB-2 gene. Eleven human lung cancer cell lines, including four derived from small cell lung cancer (SCLC) and seven derived from NSCLC were also examined for altered c-erbB-2 gene expression. Southern blot analysis of paired tumor and normal lung samples demonstrated that amplification of the c-erbB-2 gene is rare in NSCLC (2/60) and not restricted to adenocarcinomas. One patient showed an EcoRI restriction fragment length polymorphism for the c-erbB-2 locus. Four of four SCLC cell lines demonstrated minimal or nondetectable expression of c-erbB-2 mRNA compared to high levels of expression by seven of seven NSCLC lines. The highest expression levels were seen in four of four adenocarcinomas. We conclude that c-erbB-2 expression is different in SCLC compared to NSCLC and high expression of c-erbB-2 is consistently present in lung adenocarcinomas. PMID- 2569930 TI - Reversal mechanism of multidrug resistance by verapamil: direct binding of verapamil to P-glycoprotein on specific sites and transport of verapamil outward across the plasma membrane of K562/ADM cells. AB - The calcium channel blocker verapamil has been shown to reverse multidrug resistance (T. Tsuruo et al., Cancer Res. 41: 1967-1972, 1981), but the mechanism of action of this agent has not been fully elucidated. A radioactive photoactive analogue of verapamil, N-[benzoyl-3,5-3H-(+/-)-5-[(3,4 dimethoxyphenetyl)methylamino]-2- (3,4-dimethoxyphenyl)-2-isopropyl-N-p azidobenzoylpentylamine, was used to label the plasma membranes of a human myelogenous leukemia cell line (K562), a multidrug-resistant subline selected for resistance to Adriamycin (K562/ADM) and its revertant cell (R1-3). Sodium dodecyl sulfate-polyacrylamide gel electrophoretic fluorograms revealed the presence of an intensely radiolabeled Mr 170,000-180,000 protein in the membranes from K562/ADM but not from the drug-sensitive parental K562 and revertant R1-3 cells. The Mr 170,000-180,000 verapamil acceptor was immunoprecipitated by monoclonal antibody MRK16 specific for P-glycoprotein associated with multidrug resistance, indicating that P-glycoprotein in the plasma membrane is a major target of verapamil in K562/ADM cells. The photolabeling of P-glycoprotein with N-[benzoyl 3,5-3H]-(+/-)-5-[(3,4-dimethoxyphenetyl)methylamino]-2- (3,4-dimethoxyphenyl)-2 isopropyl-N-p-azidobenzoylphentylamine was significantly blocked by other calcium channel blockers, nicardipine and diltiazem, that have been shown to overcome multidrug resistance. In addition, the photolabeling was partially blocked by Adriamycin, vincristine, and colchicine, suggesting that the specific binding sites for verapamil on P-glycoprotein are closely related to the binding sites for these calcium channel blockers and antitumor agents. To determine whether verapamil could be a substrate for P-glycoprotein, the cellular accumulation of [3H]verapamil into K562 and K562/ADM was evaluated. The accumulation of [3H]verapamil in the multidrug-resistant cells was 30% of K562 cells and increased when K562/ADM cells were treated with vincristine and nicardipine at 5 microM, indicating that the P-glycoprotein transports verapamil as well as other antitumor agents in the multidrug-resistant cells. These results suggest that verapamil enhances antitumor agent retention through competition for closely related binding sites on P-glycoprotein. PMID- 2569929 TI - Biochemical pharmacology of 2-chlorodeoxyadenosine in malignant human hematopoietic cell lines and therapeutic effects of 2-bromodeoxyadenosine in drug combinations in mice. AB - Growth of human hematopoietic cell lines showed a 100-fold range of sensitivity to inhibition by 2-chloro-2'-deoxyadenosine (CldAdo), with highly sensitive lines in all three groups: T-lymphoblastic, B-lymphoblastic, and non-T, non-B. Formation of nucleotides from [8-3H]CldAdo was investigated in ten lines. In cells exposed to 0.15 microM CldAdo, CldAdo 5'-phosphate (CldAMP) reached 0.7-14 microM and CldAdo 5'-triphosphate (CldATP) reached 0.05-6 microM in 1 h. In most cases these nucleotide concentrations at 1 h were close to the steady-state concentrations, and the latter concentrations were approximately proportional to extracellular CldAdo concentration. On removal of extracellular CldAdo, intracellular CldAMP and CldATP declined rapidly with half times of 0.56-0.9 and 0.64-1.46 h, respectively. There was no correlation between these rates of catabolism and steady-state levels. The different sensitivities of the lines to CldAdo is explained only in part by the different steady-state concentrations of CldATP, and must be more directly related to differential effects on target enzymes. Mice inoculated with L1210 leukemia were treated with 2-bromo-2' deoxyadenosine (BrdAdo) paired with one of 18 other therapeutic agents. Eight of the drugs paired with BrdAdo gave therapeutic responses from the combination greater than the sum of the responses of members of the pair. They included alkylating agents, antimetabolites blocking deoxyribonucleotide synthesis, and DNA polymerase inhibitors. Toxic dosages of CldAdo caused damage chiefly to the hemic-lymphatic systems and the kidneys. PMID- 2569931 TI - Concordant deletions of chromosome 3p and loss of heterozygosity for chromosomes 13 and 17 in small cell lung carcinoma. AB - Common regions of loss of heterozygosity on chromosomes 3, 13, and 17 were determined by restriction fragment length polymorphism analysis in 34 tumors and nine cell lines from 27 patients with small cell lung carcinoma. The common regions of loss of heterozygosity on chromosomes 3, 13, and 17 reside between D3S2 (3p14-p21) and ERBA beta (3p22-p24.1), between D13S1 (13q12) and D13S2 (13q22), and distal to MYH2 (17p13.1), respectively. Allele loss in each of these regions has been previously shown in several human tumors. Thus, the present findings indicate the pleiotropy of recessive genetic lesions in these genomic areas. Cytogenetic analysis was performed on three small cell lung carcinoma cell lines which had allele loss on all three chromosomes, and although chromosome 3p deletions were observed in two of three cell lines, no obvious structural abnormalities involving chromosomes 13 and 17 were detected. Mitotic recombination or mitotic nondisjunction rather than deletion may thus be the frequent chromosomal mechanism for attaining homozygosity of chromosomes 13 and 17 in small cell lung carcinoma. PMID- 2569933 TI - Somatostatin and somatostatin analogues in oncology. PMID- 2569932 TI - Atypical multidrug resistance in a therapy-induced drug-resistant human leukemia cell line (LALW-2): resistance to Vinca alkaloids independent of P-glycoprotein. AB - Near diploid leukemic T-cells (LALW-2), exposed to cytotoxic drugs only as a consequence of therapy administered to the donor patient, have been maintained by serial xenograft in nude mice. In comparison with the leukemic line CCRF-CEM, using a growth inhibition assay, LALW-2 cells were resistant to Vinca alkaloids and actinomycin D (relative resistance, 200-fold or more), were slightly resistant to Adriamycin (relative resistance, 4-fold), and showed no resistance to daunorubicin or teniposide. By comparison, a vincristine-resistant CEM subline developed in our laboratory (CEM/VCR R) was resistant to all these agents by at least 30-fold. The VCR R subline served as a positive control, confirming the previously reported correlation between multidrug resistance and amplification of the P-glycoprotein gene. Comparison of CEM, CEM/VCR R, and LALW-2 cells establish that the P-glycoprotein gene was not amplified or overexpressed in the LALW-2 cells; neither could the gene product be detected by immunoblotting in extracts from these cells. The LALW-2 cells were further distinguished from CEM/VCR R cells due to the lack of increased vincristine efflux by the xenografted cells, an effect readily demonstrable in the CEM/VCR R cells. However, although LALW-2 cells efflux vincristine at the same rate as CCRF-CEM cells, the xenografted cells exhibited a reduced rate of vincristine accumulation. Uptake of daunorubicin by LALW-2 cells was not distinguished from that by CEM cells, consistent with similar 50% inhibitory dose levels for this drug in both cell populations, and differentiating both from CEM/VCR R cells. Thus, clinical resistance in this case appears to be an "atypical" form of multidrug resistance specifically distinguished by resistance to Vinca alkaloids and actinomycin D occurring in the absence of increased amounts of P-glycoprotein and manifesting decreased drug uptake. PMID- 2569934 TI - CD3Ti+ human thymocyte-derived clones displaying a differential response to activation via CD3Ti and CD2. AB - Previous studies indicated that, unlike peripheral T-cells, freshly isolated thymocytes show little or no proliferation to activation signals via either the antigen/MHC receptor complex (CD3Ti) or the CD2 structure, unless exogenous IL-2 or phorbol esters are added. To investigate these differences in more detail, we have studied the response of clonal populations of mature thymocyte subsets as well as peripheral T-cell clones to activation via either CD3Ti or CD2. Here we report the characterization of three clones belonging to different subsets of mature thymocytes: CD3+ CD4+ (Ti alpha/beta), CD3+ CD8+ (Ti alpha/beta), and CD3+ CD4- CD8- (Ti gamma/delta). All three clones could be induced to proliferate to insolubilized anti-CD3 mAb. In contrast, activating anti-CD2 mAbs, which induced proliferation in all peripheral T-cell clones tested, did not induce an appreciable proliferation of the thymocyte clones. The latter required additional signals provided by the phorbol ester PMA. However, anti-CD2 mAbs were able to induce early activation events such as phosphoinositide turnover and [Ca2+]i increase to an extent similar to the ones elicited by anti-CD3 mAb. These results further support previous findings suggesting that mature thymocytes are not functionally identical to peripheral T-cells. PMID- 2569935 TI - Depletion of L3T4+ and Lyt-2+ cells by rat monoclonal antibodies alters the development of adoptively transferred experimental autoimmune thyroiditis. AB - To delineate the contribution of L3T4+ and Lyt-2+ cells in the pathogenesis of experimental autoimmune thyroiditis (EAT), synergistic pairs of monoclonal antibodies (mAb) to the T cell subsets were used in conjunction with the adoptive transfer of mouse thyroglobulin (MTg)-activated cells from immunized mice. Initial experiments verified the important role of L3T4+ cells in the transfer of EAT. Subsequent experiments pointed to the relative contribution of both L3T4+ and Lyt-2+ cells, depending on the stage and extent of disease development. Treatment during disease with L3T4, but not Lyt-2, mAb alone significantly reduced thyroiditis. However, in situ analysis of the cellular infiltrate in thyroid sections revealed that, after treatment with mAb, the appropriate subset was eliminated without altering the amount of the other subset in the remaining lesion. In addition, treatment during severe thyroiditis following the transfer of MTg-activated lymph node cells showed that Lyt-2 mAb alone also reduced thyroid infiltration. When the recipients were pretreated with either pair of mAb before transfer, disease development was only moderately affected. We conclude that (i) donor L3T4+ cells are the primary cells responsible for the initial transfer and development of thyroiditis; and (ii) previous in vitro cytotoxicity data, plus current monoclonal antibody treatment of disease and in situ analysis, further implicate a role for Lyt-2+ cells in EAT pathogenesis. PMID- 2569936 TI - T cell subsets mediating lethal graft versus host disease: demonstration that synergy between CD4+ and CD8+ T cells is the predominant mechanism in low responder rat strains. AB - T cell subsets from rat strains that have been characterized as high and low responders to alloantigen were examined for their capacity to mediate lethal graft versus host disease (GVHD) across strain combinations incompatible for class I, class II, and non-MHC antigens. Inocula of 5 X 10(7) lymph node and spleen cells (LC) from low responder DA (RT1a) and high responder W/F (RT1u) strains caused lethal GVHD in (W/F X DA)F1 hybrids given 6 Gy whole body irradiation. W/F CD4+ (W3/25+) cells (2 X 10(7], equal to the number in 5 X 10(7) LC mediated lethal GVHD but 10(8) DA CD4+ cells were required to cause lethal GVHD. CD8+ (MRC OX8+) cells (5 X 10(7] from W/F rats alone caused lethal GVHD but those from DA rats could not. Mixtures of CD4+ and CD8+ DA T cells, equivalent to the number in 5 X 10(7) LC, did mediate lethal GVHD, demonstrating that synergy between the subsets was the predominant mechanism with DA cells. These results suggest that differences in alloreactivity between the strains tested may be due to alternate requirements for the alloactivation of T cell subsets; the high responder subsets being self-sufficient and the low responder subsets being dependent upon each other. PMID- 2569937 TI - Phenotypic characterization of cynomolgus monkey natural killer cells. AB - Natural killer (NK) activity of cynomolgus monkey peripheral blood lymphocytes (PBL) was determined using B95-8 cells as target cells. Examination for the reactivity of human NK-related monoclonal antibodies (mAbs), anti-Leu-7, anti-Leu 11b, anti-NKH1A, and NC-1, with cynomolgus PBL revealed that Leu-11b (CD16) was the only antigen expressed on cynomolgus PBL. The percentage of Leu-11b-positive (Leu-11b+) cells correlated well with the level of NK activity when PBL taken from 21 monkeys were tested. After depletion of Fc receptor-positive (FcR+) cells, NK activity was lost concomitantly with the disappearance of Leu-11b+ cells. These results show that cynomolgus NK cells are mainly FcR+ which can be detected by mAb directed to Leu-11b. Cynomolgus PBL were separated by Ficoll Hypaque centrifugation after E rosette formation with 2-aminoethylisothiouronium bromide-treated sheep red blood cells, and NK activities of both E rosette forming (E+) and nonforming (E-) fractions were determined. The high level of killing was observed in the E- fraction, suggesting that the majority of cynomolgus NK cells was contained in the E- fraction. The separation of PBL by Percoll discontinuous density gradient showed cynomolgus NK cells were enriched in the low density fractions. PMID- 2569938 TI - CD8 suppressor cell activity and its effect on CD4 helper cell-dependent growth of SJL/J B-cell lymphomas. AB - CD8 cells, flow cytometrically sorted from the lymph nodes of tumor-bearing and normal SJL/J mice, suppressed in vitro proliferation of syngeneic CD4 cells in response to concanavalin A, two independent SJL/J lymphomas, and LPS-activated syngeneic B-cell blasts. The data confirm earlier reports that nonspecific suppressor cells are generated as a consequence of SJL/J lymphoma-stimulated T cell proliferation. Earlier reports are extended, in that the suppressor cell is identified as expressing CD8, and the suppressor activity is shown to decrease the tumor-stimulated CD4 cell proliferation which is essential to growth of these CD4-dependent murine B-cell lymphomas. In three separate experiments, anti-CD8 treatment of mice, in which CD4 cells were made limiting by injection with anti CD4, increased growth of transplantable SJL/J lymphomas with corresponding increases in numbers of CD4 cells. The data imply that, under certain conditions, CD8 suppressor cells measurably influence growth of SJL/J lymphomas by regulating the tumor-stimulated CD4 cell proliferation essential to maximum growth of SJL/J lymphomas. PMID- 2569939 TI - Long-term growth of human WGA-activated T-lymphocytes without feeder cells. AB - Human wheat germ agglutinin (WGA)-activated T-lymphocytes from either peripheral blood or spleen could be propagated for several weeks in the presence of culture supernatants from human mononuclear cells obtained after a 1-day stimulation with WGA and after a subsequent 3-day culture without mitogen. The continuous T-cell growth in this culture system required alternate exposition of the cells to the above supernatants, both with and without WGA, but proceeded in the absence of feeder cells. After 3 weeks of propagation most cells displayed the CD4+ phenotype, expressed IL2 receptors, and responded to PHA, Con A, and WGA. It has been shown that WGA-activated T-cells could be cloned by limiting dilution and propagated using the above culture system. PMID- 2569941 TI - Synthesis and analgesic effect of normorphine-3- and -6-glucuronides. AB - Normorphine-3- and -6-glucuronides were synthesized, and their analgesic effects were examined. Normorphine-3-glucuronide was obtained by condensation of normorphine with acetobromoglucuronate in the presence of sodium hydroxide in acetone. On the other hand, normorphine-6-glucuronide was synthesized by condensing N,O3-biscarbobenzoxynormorphine with acetobromoglucuronate in the presence of silver carbonate, and removing the protecting groups from the resultant reaction product by catalytic hydrogenation and solvolysis with sodium methoxide and barium hydroxide. The analgesic effect of normorphine-6-glucuronide (ED50 0.036 nmol/mice) was 125-fold more potent than that of normorphine in mice injected i.c.v. Normorphine-3-glucuronide was shown to be 37% effective at a dose of 2 nmol/mice, but induced convulsion at higher doses when given by i.c.v. injection. PMID- 2569940 TI - The control of aromatic amino acid catabolism and its relationship to neurotransmitter amine synthesis. AB - The aromatic amino acids are, inter alia, substrates for the synthesis of important neurotransmitters. Although the factors controlling the synthesis of these transmitters are not fully understood, there is evidence that the concentrations, both relative and absolute, of the precursor amino acids in the blood are of some significance. The article reviews the biochemical pathways involved in tryptophan, phenylalanine, and tyrosine metabolism in liver, brain, and other tissues and discusses (1) the major regulatory events in the maintenance of blood concentrations and (2) the effects of diet, load dosing, hormones, and other circulating substances on the fate of the amino acids and on events in the central nervous system. PMID- 2569943 TI - Quantitative structure-activity relationships of H1-antihistaminic benzimidazole derivatives. AB - The quantitative structure-activity relationships (QSAR) of 2-(4-substituted-1 piperazinyl)benzimidazole derivatives for antihistaminic activity were examined. Taking into consideration the specific conformations of some derivatives, a significant correlation was obtained using Verloop's STERIMOL parameters B3 and L of the substituent at the 1-position of the benzimidazole nucleus. The results indicated that the derivatives having a substituent with a small breadth and an appropriate length at the 1-position showed potent activity. From the results, a model of the binding site is proposed. The QSAR of side effects (anticholinergic activity and central nervous system depressive effect) were also examined and the results showed that a sterically small substituent at the 1-position was required to decrease side effects. PMID- 2569942 TI - Syntheses of the metabolites of 1-(2-ethoxyethyl)-2-(hexahydro-4-methyl-1H-1,4 diazepin-1-yl)-1H benzimidazole difumarate (KG-2413) and related compounds. AB - The metabolites of 1-(2-ethoxyethyl)-2-(hexahydro-4-methyl-1H-1,4-diazepin-1-yl) 1H-b enzimidazole difumarate (KG-2413), which has a potent H1-antihistaminic activity, were predicted on the basis of metabolic studies of related compounds and were synthesized to aid in identification of the actual metabolites and for examination of their antihistaminic activity. Among the twelve compounds prepared, nine compounds were actually found as the metabolites of KG-2413 in rat urine. The antihistaminic activities of these metabolites were found to be lower than that of KG-2413. PMID- 2569944 TI - The food-mutagens 2-amino-1-methyl-6-phenylimidazo-[4,5-b]-pyridine (PhIP) and 2 amino-3,8-dimethylimidazo[4,5-f]-quinoxaline (MeIQx) initiate enzyme-altered hepatic foci in the resistant hepatocyte model. AB - The initiating activity of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and a mutagenic meat extract obtained from cooked meat was examined, using the resistant hepatocyte model (RH-model). Male Wistar rats were given a single i.p. injection of PhIP (50 or 75 mg/kg body weight) or MeIQx (50 mg/kg body weight) after a 2/3 partial hepatectomy (PH). The meat extract (corresponding to 1050 g meat/animal) was given by gastric feeding at three time points after PH. Two weeks after initiation the rats received a diet containing 0.02% 2-acetylaminofluorene for a period of 2 weeks. In the middle of this period a single dose of carbon tetrachloride was given. Rats were killed 6 weeks after the experimental start. The number of enzyme-altered (gamma-glutamyl-transferase positive) hepatic foci was significantly increased in the animals given MeIQx (P less than 0.05) and the highest dose of PhIP (P less than 0.01) whereas no effect of the meat extract was observed. The mutagenic meat extract was also studied with regard to promotive capacity in vivo, the meat extract was given in the diet of diethylnitrosamine initiated rats for a period of 6 weeks. No significant changes were detected after administration of the meat extract in the diet. It is concluded that both MeIQx and PhIP are weak initiators in the RH-model, in the same order of magnitude as the previously investigated, structurally related pyrolysis products. The meat extract was not active in the RH-model under the conditions used in this study. PMID- 2569945 TI - Regulation of cell proliferation by beta-adrenergic receptors in a human lung adenocarcinoma cell line. AB - We have recently reported that the tobacco-related nitrosamines N nitrosodiethylamine (DEN) and 4-(methyl-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK) stimulate cell proliferation in cell lines derived from human neuroendocrine carcinoma and adenocarcinoma (comprised of Clara cells) of the lung. In the neuroendocrine cell line, this effect was inhibited by antagonists of nicotinic cholinergic receptors which regulate the secretion of peptide hormones and cell proliferation of pulmonary neuroendocrine cells. No such inhibition was observed in the adenocarcinoma line. Clara cells reportedly do not have acetylcholine receptors and secretion of this cell type is regulated by beta adrenergic receptors instead. In this experiment, we test the hypothesis that the latter types of receptors are involved in the regulation of cell growth of normal and nitrosamine-stimulated adenocarcinoma cells with features of Clara cells. Our data demonstrate a pronounced stimulation of cell growth by the beta-adrenergic agonist isoproterenol, DEN and NNK, as well as a dose-dependent inhibition of such growth-stimulating effects by the beta-adrenergic antagonist propranolol. These findings suggest an important role of beta-adrenergic receptors in the regulation of cell proliferation in lung tumors comprised of this cell type. PMID- 2569946 TI - Natural history of aortoarteritis (Takayasu's disease). AB - The natural history of aortoarteritis was studied in 88 patients (54 women and 34 men). The average age was 24.0 +/- 8.8 years at onset of symptoms and 28.3 +/- 9.9 years at diagnosis. The follow-up period was 83.6 +/- 74.4 months from onset and 33.2 +/- 37.0 months from diagnosis. Ten patients (11.4%) died during follow up (0.016 deaths/patient year), and 22 patients (25%) suffered major nonfatal events (0.042 events/patient year). The cumulative survival at 5 and 10 years after the onset was 91.0 +/- 3.3% and 84.0 +/- 5.6% (mean +/- SEM), respectively. The event-free survival rates at the same intervals after onset were 74.9 +/- 5.0% and 64.0 +/- 7.4%, respectively. The overall survival and event-free survival at 10 years after diagnosis was 80.3 +/- 6.5% and 61.6 +/- 7.5%, respectively. Patients with no complications or a mild single complication at diagnosis had a higher event-free survival rate than those with severe single complication or multiple complications at 5 years--97.0 +/- 2.9% and 59.7 +/- 7.3%, respectively (p less than 0.001). Severe hypertension (p less than 0.01), severe functional disability (p less than 0.01), and evidence of cardiac involvement (p less than 0.05) were good predictors of either death or major event on follow-up. These data are useful in making an objective assessment of the prognosis and in planning elective interventions. PMID- 2569947 TI - Persistence of sympathetic-mediated forearm vasoconstriction after alpha-blockade in hypertensive patients. AB - Sympathetic vasoconstriction not mediated by alpha-adrenoceptors has been identified in vitro and in animals but not in humans. We evaluated the effect of alpha-adrenoceptor blockade on either endogenous vascular sympathetic activation (obtained through the application of a nonhypotensive lower-body negative pressure, -10 mm Hg for 5 minutes) or selective postsynaptic alpha-adrenoceptor stimulation by exogenous norepinephrine (0.005 micrograms/100 ml forearm tissue/min for 3 minutes) in the presence of beta-blockade by propranolol (10 micrograms/100 ml forearm tissue/min for 15 minutes). Drugs were infused into the brachial artery at systemically ineffective rates while continuously monitoring forearm blood flow (by venous plethysmography), intra-arterial mean arterial pressure, and heart rate in patients with essential hypertension. The irreversible antagonist phenoxybenzamine was used at a rate of 20 micrograms/100 ml forearm tissue/min for 1 hour, which antagonized the local responses to norepinephrine in a range of 0.005-0.05 micrograms/100 ml forearm tissue/min. During saline administration, either lower-body negative pressure or exogenous norepinephrine decreased forearm blood flow comparably. However, after phenoxybenzamine administration, forearm vasoconstriction to norepinephrine was abolished while a residual response to lower-body negative pressure remained in each patient. To exclude insufficient alpha-adrenoceptor blockade, the same experimental protocol was repeated by doubling phenoxybenzamine concentrations. No difference from the data obtained with the lower level of antagonist was found. Further studies were performed to confirm the sympathetic origin of the residual vasoconstriction. Bretylium tosylate, a neurotransmitter blocker, infused into the brachial artery (50 micrograms/100 ml forearm tissue/min for 90 minutes) abolished the effect of endogenous sympathetic activation but did not alter the effect of exogenous norepinephrine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569949 TI - Haplotype analysis of the phenylalanine hydroxylase gene in Turkish phenylketonuria families. AB - We have estimated the haplotype distribution of mutant and normal phenylalanine hydroxylase (PAH) alleles for 17 Turkish phenylketonuria (PKU) families: 20 normal and 27 mutated PAH alleles could be identified. Of the latter, the most prevalent were associated with haplotype 6 (29.6%), 1 (18.5%) and 36 (11.1%), while the normal alleles were preferentially associated with haplotype 1 (20%). Of the 19 different haplotypes observed, 5 have not been described previously. The haplotype distribution differed significantly from that of the Northern European population. Two of the eight polymorphic sites were in association with PKU. No deletions of exon sequences were found in the families analysed. PMID- 2569948 TI - Surgical consideration of aortitis involving the aortic root. AB - Most serious complications after aortic valve replacement (AVR) in Takayasu's and Behcet's diseases are repeated detachment of the prosthetic valve and formation of a false aneurysm. Therefore, special consideration of the surgical treatment of aortitis involving the aortic root is required. Fifteen patients (12 with Takayasu's disease and three with Behcet's disease) underwent various surgical procedures, depending on the pathological changes in the aortic valve and the aortic root and on the degree of involvement of the coronary artery. Group 1 patients had coronary artery involvement alone and underwent coronary artery bypass grafting (CABG) (n = 3); group 2 patients had aortic regurgitation with an intact coronary artery and underwent AVR or a modified Bentall procedure (n = 7); and group 3 patients had aortic regurgitation with coronary artery involvement and underwent AVR, a modified Bentall procedure, or translocation with CABG (n = 5). A prosthetic valve or composite graft with a Teflon felt flange at the sewing ring was fabricated during surgery by exact measurement of the sizes of the aortic annulus and distal aorta and treatment with fibrin glue before insertion. The double-fixation method with reinforcement by a Teflon felt strip was employed for anastomosis of the flanged prosthesis. The button-shaped coronary ostium was directly anastomosed to the composite graft. One patient required translocation with CABG because of a deteriorated annulus. There were no operative or hospital deaths. One patient died of a brain abscess at 6 months after surgery, and another patient who underwent CABG required CABG reoperation due to graft occlusion. We recommend postoperative steroid therapy in patients who are diagnosed as being in the active stage of the disease until inflammatory signs disappear. PMID- 2569951 TI - Does suppression of responsiveness to beta-adrenoceptor activation explain the enhancement of vasoconstrictor responses by alpha 2-adrenoceptor agonists? AB - 1. The alpha 2-adrenoceptor agonist TL99, in concentrations that had no other observable effect, enhanced constrictor responses to phenylephrine in perfused segments of the rat tail artery. 2. Vasoconstrictor responses to phenylephrine were also enhanced by propranolol (0.3 mumol/L). 3. Vasoconstrictor responses to phenylephrine in the presence of propranolol were further enhanced by TL99 (10 nmol/L). 4. The enhancement of vasoconstrictor responses by alpha 2-adrenoceptor agonists is not due to the removal of a counteracting vasodilator component produced by activation of beta-adrenoceptors. PMID- 2569952 TI - Dopamine receptors: classification, properties and drug development. AB - 1. The pharmacology, biochemistry and structure-activity relationships pertinent to the D1 and D2 subtypes of dopamine receptors are reviewed. 2. Recent advances in receptor purification and our understanding of the secondary messenger systems involved are presented. 3. D1 and D2 receptors of central nervous, endocrine and peripheral systems are identical. PMID- 2569950 TI - Thyroglobulin antibodies in Graves' disease are associated with T-cell receptor beta chain and major histocompatibility complex loci. AB - We have investigated the T-cell antigen receptor constant beta and alpha chain genes (TCR-C beta, -C alpha) and the immunoglobulin (Ig) heavy chain switch regions of patients with Graves' disease (GD) using restriction fragment length polymorphism (RFLP) analysis. No significant associations were found with RFLPs of either the TCR-C beta, -C alpha or Ig heavy chain switch region loci and GD. However, a significant association was found between the presence of anti thyroglobulin (anti-Tg) antibodies in the serum of patients and the 10.0; 9.2 kb TCR-C beta genotype (P less than 0.02). Also, those patients with anti-Tg antibodies had an increased frequency of HLA-DR3 (P less than 0.025). These results suggest that genes residing in the TCR chain and major histocompatibility complex loci may be important in determining the immune response to thyroglobulin but not to the disease itself. PMID- 2569953 TI - Function, characterization and autoradiographic localization and quantitation of beta-adrenoceptors in cardiac tissues. AB - 1. This paper demonstrates the use of organ bath, radioligand binding and autoradiography to detect beta 1- and beta 2-adrenoceptors in human and guinea pig cardiac tissues. 2. In organ bath experiments, non-selective and beta 1- and beta 2-adrenoceptor selective agonists produced concentration-dependent inotropic responses in human right atrial appendage. Both subtypes mediate inotropic responses. In guinea-pig right atria chronotropic responses were mediated predominantly through beta 1-adrenoceptors. 3. Receptor binding studies using ( )[125I]-cyanopindolol (CYP) and beta 1- and beta 2-adrenoceptor selective antagonists showed that beta 2-adrenoceptors comprised 25% of the total population of beta-adrenoceptors in guinea-pig right atria. In human right atria the proportion is higher (40%). 4. Quantitative autoradiography was used to determine the location and densities of beta 1- and beta 2-adrenoceptors in guinea-pig heart. Both beta 1- and beta 2-adrenoceptors were distributed on myocardium. The atrioventricular conducting system had a higher density of beta 2 adrenoceptors compared with myocardium. PMID- 2569954 TI - Interleukin 2 responsive lymphocytes in patients with adenosine deaminase deficiency. AB - We evaluated the effects of recombinant interleukin 2 (IL-2) on the proliferative responses to mitogens of peripheral blood mononuclear cells (PBMC) from three adenosine deaminase (ADA)-deficient patients. There was significant enhancement by IL-2 of the proliferative responses to phytohemagglutinin (PHA) and pokeweed mitogen (PWM) of PBMC from all three patients. We found that normal PBMC respond with increased numbers of CD3-positive cells when exposed to PHA or PWM and that the response by normal CD8-positive cells was greater than that by CD4-positive cells. In contrast, we found that in ADA-deficient cells the response is almost entirely due to the CD3/CD4-positive population of lymphocytes. These results could not be explained by either the culture conditions or the possibility of a mixed chimeric state. When we evaluated an in vitro cell model of ADA deficiency using an ADA inhibitor, erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA), we found that the inhibitory effect of EHNA plus deoxyadenosine on mitogen-stimulated PBMC could not be prevented by IL-2. These results suggest that the immunodeficiency in ADA deficiency includes the absence or failure of a subset of T cells to make IL-2 and the failure of the CD8-positive subset to respond to IL-2. Also, the in vitro cell model of ADA deficiency using EHNA as the ADA inhibitor is limited in its use in understanding the pathogenesis of this disease. PMID- 2569956 TI - [Treatment of progressive supranuclear palsy]. AB - The neuropharmacological and neurochemical features of Progressive Supranuclear Palsy (PSP) are reviewed, together with the results obtained by various therapeutic trials. PSP is a neurodegenerative disease which often causes Parkinsonian symptoms but dopaminomimetic drugs have given rise to poor improvement, in spite of dopamine decrease observed in PSP patients' nigrostriatal region. The uselessness of L-DOPA therapy in PSP patients may explain the numerous failures encountered in PSP patients misdiagnosed as Parkinsonian. On the basis of the recent discovery of striatal dopaminergic receptor abnormalities and of interaction between various neurotransmitters, the authors suggest some possible therapeutic substances that might improve the outcome of the disease. PMID- 2569955 TI - Monoclonal antibodies to CD18 and CD11A (LFA-1) distinguish Down's syndrome (trisomy 21) from normal lymphoblastoid cells. AB - To evaluate the potential of using fluorescence flow cytometry (FFC) in the diagnosis of Down's syndrome we analysed a series of Trisomy 21 lymphoid cell lines (LCL) with a panel of monoclonal antibodies (MoAb) to the CD11a and CD18 subunits of lymphocyte function associated antigen 1 (LFA-1). In pairwise analysis of Trisomy 21 (T21) and normal LCL with 6 CD18 and 10 CD11a MoAb we found that T21 LCL could be distinguished from normal LCL with both types of MoAb, even though only the CD18 gene is duplicated in Down's syndrome. Experiments in which CD18 or CD11a were capped showed that the two subunits co migrate on T21 and normal LCL, probably as heterodimers. This appears to explain the increased expression of CD11a on Trisomy 21 LCL. There was no evidence that T21 LCL express the other two leucocyte integrin subunits CD11b and CD11c, indicating that Trisomy 21 has little affect on tissue specific control of these molecules. In contrast, there appears to be reduced expression of an unrelated adhesion molecule LFA-3 on Trisomy 21 LCL. We discuss the relevance of these results in the diagnosis of Down's syndrome. PMID- 2569958 TI - Urinary angiotensin-I-converting enzyme activity as a marker of tubulo interstitial involvement in kidney diseases. PMID- 2569957 TI - Concentrations of oestradiol and testosterone in peripheral and spermatic venous blood of dogs with unilateral cryptorchidism. AB - Plasma testosterone and oestradiol concentrations were measured in peripheral and spermatic venous blood of 13 dogs with unilateral inguinal cryptorchidism, 9 dogs with unilateral abdominal cryptorchidism, and in a control group of 36 mature normal dogs. The hormone concentrations were similar in the three groups, both in the peripheral and in the spermatic venous blood. The weight of the testes in the control group was correlated with the body weight and there was no significant difference between the weight of the right and the left testes. The weight of the abdominal testes was lower than that of the inguinal testes, and there was no compensatory enlargement of the contralateral scrotal testis. The cryptorchid testes showed little or no histological evidence of spermatogenesis, and spermatogenesis was usually normal in the scrotal testes. PMID- 2569960 TI - Almitrine decreases the distensibility of the large pulmonary arteries in man. AB - Almitrine improves blood gas values in patients with COPD, primarily through better ventilation-perfusion matching. The improvement comes largely from a vasomotor effect of the drug. The vasomotor mechanism is unknown. We suspected that decreased distensibility of the large pulmonary arteries was one probable effect of the drug. If present, reduced distensibility would explain many of the hemodynamic alterations produced by almitrine. To test this idea, we measured the diameter of the right pulmonary artery during systole and diastole both before and after almitrine administration in nine patients who were undergoing pulmonary cineangiography. Our calculations of the distensibility of the right pulmonary artery showed that almitrine caused a 60 percent decrease. This significant alteration in the stiffness of the large pulmonary arteries can account for the rise in systolic pressure, a change that would be expected to redistribute pulmonary blood flow upwards and thereby improve ventilation-perfusion balance. PMID- 2569961 TI - Azelastine inhibits agonist-induced electromechanical activity in canine tracheal muscle. AB - We examined the effects of a new antiasthmatic drug, azelastine, on the electromechanical responses of airway smooth muscle to histamine, acetylcholine (ACh), and tetraethylammonium (TEA). Membrane potential and isometric force were simultaneously measured in isolated canine tracheal muscle using intracellular microelectrodes and a microforce transducer. Azelastine, at 1 microM, depressed the histamine-induced contractile force by more than 60 percent. The histamine induced membrane depolarization was also inhibited, but to a much less extent, compared to the inhibition of contractile force. Similarly, contraction induced by ACh was inhibited by azelastine. In contrast to histamine, ACh elicited electrical oscillations concomitant with the membrane depolarization. Azelastine abolished these oscillations without affecting the depolarization. Azelastine inhibited the Ca2+-dependent slow action potentials (induced by 20 mM TEA) in a concentration-dependent manner; complete inhibition occurred at 30 microM. Such direct inhibitory effects of azelastine on agonist-induced airway muscle contraction may explain its ability to exert bronchodilatation in asthmatic patients. One of its mechanisms of action may involve inhibition of voltage sensitive Ca2+ influx across the muscle cell membrane; however, additional actions intracellularly are possible. PMID- 2569959 TI - Treatment of severe hypertension with atenolol and betaxolol with once-daily regimens. Hemodynamic aspects. AB - The effectiveness and safety of once-daily administration of drugs in the treatment of moderate to severe hypertension was studied. Forty men taking diuretics were randomized to atenolol (A, n = 18), 50 mg/day, or betaxolol (B, n = 22), a new B1-blocker, 20 mg/day, if their SDAP was 105 to 125 mm Hg at baseline (weeks 2 to 4). At week 6, if SDAP was greater than 95 mm Hg, minoxidil (M), 5.5 mg/day, was added. The patients were seen every two weeks to week 16 (end of drug titration) and then every four weeks to week 32. The dosages were increased to 200 mg/day for A, 80 mg/day for B, and 20 mg/day for M as needed. Physical examinations, chest x-ray films, ECGs, echocardiograms, spirometric studies, 24-h ambulatory arterial pressures (AAP), and blood chemistry analyses were done at baseline and during treatment. A and B combined with a diuretic (furosemide, F) and M decreased the arterial pressures and heart rates equally well by both clinical and AAP measurements (p less than .001). The IVS was decreased (p less than .05), whereas LVIDd, RVIDd, and cardiothoracic ratios were increased by both A and B (p less than .05, p less than .01). No changes were noted in LVPW, LVM, EF, FS, spirometric values, or blood chemistry analyses. Common side effects were weight gain, edema, and hypertrichosis. Once-daily administration of A or B in combination with F and M were effective in the treatment of moderate to severe hypertension. Although effective, prolonged use of M may lead to volume overload and cardiomegaly. The significance of these latter findings is not yet known. PMID- 2569962 TI - Role of beta-receptor in the radix Angelicae sinensis attenuated hypoxic pulmonary hypertension in rats. AB - Acute pulmonary hypertension was caused by inhalation of 5% O2 in rats. Pulmonary vascular resistance (PVR) increased, but heart rate (HR), cardiac output (CO) and carotid arterial pressure (CAP) were not obviously changed. After an intravenous administration of Radix Angelicae sinensis, the acute pulmonary hypertension induced by inhalation of 5% O2 could be attenuated, but this effect disappeared if propranolol was given before Radix Angelicae sinensis. In chronic experiments, the same results were obtained, but the protective effect of Radix Angelicae sinensis on heart function was not influenced by propranolol. It is suggested that Radix Angelicae sinensis might play a role by stimulating the beta 2 receptor in the prevention of acute and chronic hypoxic pulmonary hypertension. However, prevention of hypertrophy of the right ventricle and enhancement of heart function in chronic hypoxic rats might not be attributed to the beta 1 receptor in the heart. PMID- 2569964 TI - [A registry of medullary thyroid carcinoma]. PMID- 2569963 TI - Results of second trimester prenatal diagnosis of cystic fibrosis in risk families. AB - Investigation of 17 children delivered after prenatal examination of amniotic fluid GGT was performed. GGT testing was carried out in the 17th-18th week of gestation. The development of children unaffected by cystic fibrosis was predicted. In all pregnancies, clinically healthy children with normal sweat chloride concentrations were delivered. Our results confirm the advantage of the examination of microvillar enzymes in amniotic fluid in the second trimester as a rather reliable method of fetal diagnosis of cystic fibrosis if it is impossible to use the molecular genetic methods in the first trimester. PMID- 2569965 TI - [Treatment of an undescended testis with human chorionic gonadotropin (HCG)]. AB - Seventy one patients with ectopic testis of age between 2 and 11.5 years were treated with human chorionic gonadotropin (HCG) at doses recommended by the International Health Foundation. The descent of testis to the scrotum was achieved in almost half of the treated boys (49.3%). The descent was successful mainly in cases of lower inguinal position of the undescended testicle, and only rarely when the testicle was situated higher. The descent was never successful in cases when the scrotum was small and underdeveloped. PMID- 2569966 TI - The use of denaturing gradient gel electrophoresis to screen for DNA sequence polymorphisms in the human factor VIII gene. AB - We describe the use of denaturing gradient gel electrophoresis to screen for DNA sequence polymorphisms in the human factor VIII gene. DNA fragments that differ in sequence by only a single base pair can be separated on denaturing gradient gels due to changes in their melting behavior. Previous studies have demonstrated the use of denaturing gradient gels to detect sequence changes in human genomic DNA, including mutations in the beta globin gene and polymorphisms on chromosome 20. We have begun to use denaturing gradient gels to look for polymorphisms within the human factor VIII gene. The DNA sequences of seven cloned fragments from introns in the human factor VIII gene were determined and used to predict a melting map for each fragment. The melting behavior of each cloned fragment was evaluated by electrophoresis into denaturing gradient gels. Appropriate fragments were then used as radioactive probes for hybridization to human DNA samples that had been digested with restriction enzymes. Heteroduplexes formed between the probe and genomic DNA samples were electrophoresed into denaturing gradient gels. The final positions of heteroduplex bands were determined by autoradiography. We describe a general approach for using denaturing gradient gel electrophoresis to find DNA polymorphisms, with particular emphasis on the predictive value of DNA sequence data. We compare the efficiency of polymorphism detection by denaturing gradient gel electrophoresis with detection by restriction fragment length polymorphism (RFLP) analysis. The factor VIII gene appears to have a low level of DNA sequence polymorphism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569967 TI - Human atrial natriuretic peptide receptor defines a new paradigm for second messenger signal transduction. AB - We isolated cDNAs encoding a 115 kd human atrial natriuretic peptide (alpha ANP) receptor (ANP-A receptor) that possesses guanylate cyclase activity, by low stringency hybridization with sea urchin Arbacia punctulata membrane guanylate cyclase probes. The human ANP-A receptor has a 32 residue signal sequence followed by a 441 residue extracellular domain homologous to the 60 kd ANP-C receptor. A 21 residue transmembrane domain precedes a 568 residue cytoplasmic domain with homology to the protein kinase family and to a subunit of the soluble guanylate cyclase. COS-7 cells transfected with an ANP-A receptor expression vector displayed specific [125I]alpha ANP binding, and exhibited alpha ANP stimulated cGMP production. These data demonstrate a new paradigm of cellular signal transduction where extracellular ligand binding allosterically regulates cyclic nucleotide second-messenger production by a receptor cytoplasmic catalytic domain. PMID- 2569968 TI - Identification and electron microscopic analysis of a chaperonin oligomer from Neurospora crassa mitochondria. AB - A 7-fold symmetric particle has been identified in Neurospora crassa which is most probably the mitochondrial chaperonin. The particle, about 12 nm in diameter, appears in preparations of cytochrome reductase, and is shown to contain a 60 kd protein which cross-reacts with anti-GroEL antibodies. Results of STEM mass measurement suggest that the particle is composed of 14 subunits. A preliminary interpretation of the structure of the particle based on electron microscopy is given. Its quaternary structure and molecular weight are similar to those of the recently discovered family of particles called chaperonins, found in bacteria, chloroplasts and mitochondria. PMID- 2569969 TI - The structural and functional organization of the murine HOX gene family resembles that of Drosophila homeotic genes. AB - This paper reports the cloning of the fourth major murine homeogene complex, HOX 5. The partial characterization of this gene cluster revealed the presence of two novel genes (Hox-5.2, Hox-5.3) located at the 5' extremity of this complex. In situ hybridization experiments showed that these two genes are transcribed in very posterior domains during embryonic and foetal development. We also show that Hox-1.6, the gene located at the 3' most position in the HOX-1 complex, has a very anterior expression boundary during early development. These results clearly support the recently proposed hypothesis that the expression of murine Antp-like homeobox-containing genes along the antero-posterior developing body axis follows a positional hierarchy which reflects their respective physical positions within the HOX clusters, similar to that which is found for the Drosophila homeotic genes. Such a structural and functional organization is likely conserved in most vertebrates. Moreover, on the basis of sequence comparisons, we propose that the ordering of homeobox-containing genes within clusters has been conserved between Drosophila and the house mouse. Thus, very different body plans might be achieved, both in insects and vertebrates, by evolutionarily conserved gene networks possibly displaying similar regulatory interactions. PMID- 2569970 TI - Two gene members of the murine HOX-5 complex show regional and cell-type specific expression in developing limbs and gonads. AB - This study reports the expression domains of two murine HOX gene members of the HOX-5 complex (Hox-5.2, Hox-5.3). These two genes have very similar homeodomain sequences, as well as temporal and spatial specificities of expression. They are both expressed at very posterior levels in the central nervous system, in sclerotome derivatives and in a few internal organs. In addition to these expression domains which are shared with other HOX genes, transcripts from both Hox-5.2 and Hox-5.3 are present at high levels in developing limbs. After an early homogeneous expression in mesodermal limb bud cells, transcription becomes restricted to cartilage-differentiating cells. In addition, Hox-5.2 is a marker for gonadal development. The possible involvement of such genes during inductive processes or organogenesis is discussed. PMID- 2569972 TI - Mutagenicity of some alkyl nitrites used as recreational drugs. AB - When the AIDS epidemic was in its earliest stages, and prior to identification of HIV as the etiological factor, the use of volatile nitrites by the male homosexual community to enhance sexual activities appeared to have a significant role in this disease. Preliminary observations indicated that the portion of the male homosexual community which developed Kaposi's sarcoma were also heavy nitrite users. These nitrites had been demonstrated to be mutagenic in bacteria and thus it was postulated that they could be responsible for the appearance of the sarcoma. To evaluate further the genotoxic activity of these chemicals, six nitrites, including those most commonly used by homosexuals for sexual gratification, were selected for testing in the mouse lymphoma TK+/- and Salmonella typhimurium mutagenicity assays. One chemical, n-amyl nitrite, was negative in the mouse lymphoma assay, while the other five chemicals, n-butyl, isobutyl, iso-amyl, sec-butyl, and n-propyl nitrite, were positive. All six compounds were positive in the Salmonella assay. The mutagenic and known toxic effects of these chemicals remain a concern because a large population of teenagers and young adults continue to abuse these substances. PMID- 2569973 TI - Human immunodeficiency virus type 2 in Spain. PMID- 2569971 TI - Spatial regulation of the Antennapedia and Ultrabithorax homeotic genes during Drosophila early development. AB - Both maternally supplied products and zygotically acting segmentation genes are required to establish the segment pattern of the Drosophila embryo. These genes are thought to act in part by regulating the expression of the homeotic genes. Products of the maternal and zygotic gap genes are present in the egg prior to blastoderm formation, when the homeotic genes are initially expressed within precisely bounded domains. In order to assess the first regulatory interactions between some of these gap gene products and the homeotic genes, we have examined the spatial distribution of transcripts arising from the homeotic Antp and Ubx genes during early embryogenesis in various mutant backgrounds. Here we show that mutations in both maternally and zygotically acting gap genes differentially affect the initial spatial domains of transcripts arising from each of these homeotic gene promoters. Later in embryogenesis, the patterns of homeotic gene expression change in both the wild-type and mutant cases, suggesting that other regulatory activities come into play. We propose a model in which the initial activation of each homeotic gene promoter depends on a unique combination of gap and pair-rule gene activities. PMID- 2569974 TI - Pharmacology of the benzodiazepine receptor. AB - The benzodiazepine receptor (BZR) is an intrinsic allosteric modulatory site of the GABAA-receptor-chloride channel complex of neuronal membranes mediating the main action of the major inhibitory neurotransmitter GABA. The BZR is unique in recognizing three classes of ligands, two of them producing opposite modulatory effects on the GABAA receptor function in an allosteric fashion (agonists and inverse agonists) and the third acting as antagonists of the two others. Agonists and partial agonists of the BZR (belonging to various chemical classes) have therapeutic applications as broad-spectrum tranquilizers and specific anxiolytics anticonvulsants, respectively. The BZR antagonist flumazenil, recently introduced in therapy, increases the versatility of agonists in therapy and greatly simplifies the treatment of agonist overdosing. Inverse agonists are interesting probes to analyse the biological basis of anxiety-related emotional disorders. PMID- 2569976 TI - Indirect evidence for separate vesicular neuronal origins of norepinephrine and ATP in the rabbit vas deferens. AB - Various modulators of neurotransmission were examined for selective effects on the non-adrenergic or adrenergic components of neurotransmission in the vas deferens to test the hypothesis that ATP and norepinephrine are secreted from the same vesicles. The ATP receptor antagonist, arylazido aminopropionyl ATP (ANAPP), selectively depressed the non-adrenergic contraction and prazosin selectively depressed the adrenergic contraction in response to electrical stimulation. These results are consistent with the presence of two neurotransmitters, ATP and norepinephrine, which mediate neurogenic contractions. Prostaglandin E2 inhibited non-adrenergic, but enhanced adrenergic, electrically induced (10 Hz) contractions, presumably via a prejunctional mechanism. The adrenergic component of the neurogenic response was significantly more sensitive to treatment with guanethidine, guanabenz, and 6-hydroxy-dopamine. These results with a variety of agents are inconsistent with the hypothesis that the neurotransmitters. ATP and norepinephrine, are released in tandem from the same neuronal granules. PMID- 2569977 TI - Acute and long-term regulation of brain alpha 2-adrenoceptors after manipulation of noradrenergic transmission in the rat. AB - The specific binding of [3H]clonidine (KD and Bmax) to rat brain membranes was used as a biochemical index to directly evaluate alpha 2-adrenoceptor changes after manipulation of synaptic noradrenaline (NA) pools or stimulation or blockade of the receptor. Acute (2 h) and prolonged (7 days) inhibition of NA synthesis with alpha-methyl-p-tyrosine (150 mg/kg) or acute (2 h) and chronic (14 days) treatment with reserpine (0.1-0.5 mg/kg) reduced the NA content by 15-90%, which also resulted in marked reductions (35-55%) of the KD values for [3H]clonidine in all brain regions studied. In contrast to alpha-methyl-p tyrosine, chronic reserpine treatment did not alter the Bmax values for [3H]clonidine or [3H]UK 14304 in any brain region. In the hypothalamus and cerebral cortex, acute (2 h) and chronic (7-14 days) treatment with the monoamine oxidase (MAO) inhibitors clorgyline (1 mg/kg) or tranylcypromine (5 mg/kg) increased the content of NA by 6-100%, which led to marked reductions (20-50%) of Bmax without altering the KD values for [3H]clonidine. Similarly, prolonged (21 days) inhibition of NA neuronal uptake with cocaine or protriptyline (10 mg/kg) also resulted in decreases in Bmax (20-25%) with no alterations in KD in the hypothalamus. In various brain regions, chronic (14 days) but not short-term (1 day) treatment with clonidine (0.1 mg/kg) or yohimbine (10 mg/kg) resulted in decreases (30-40%) and increases (15-20%), respectively, in Bmax without altering the KD values for [3H]clonidine. The results indicate that drugs which deplete endogenous NA up-regulate alpha 2-adrenoceptors (increased affinity of [3H]clonidine binding sites) while drugs which increase the intraneuronal and/or synaptic NA pools down-regulate the receptors (decreased number of [3H]clonidine binding sites). These adaptive receptor changes appear to be dependent on NA availability. PMID- 2569975 TI - Clozapine: new research on efficacy and mechanism of action. AB - Clozapine can produce greater clinical improvement in both positive and negative symptoms than typical antipsychotic drugs in neuroleptic-resistant schizophrenic patients. The clinical response may occur rapidly in some patients but is delayed in others. Clozapine has also been reported to produce fewer parkinsonian symptoms, to involve a lower risk of producing tardive dyskinesia, and to produce no serum prolactin elevations in man. It seems likely that these effects are the result of a common biological mechanism or related mechanisms, rather than unrelated effects. Other atypical antipsychotic drugs, such as melperone and fluperlapine, share at least some of these properties. A relatively low affinity for the D-2 dopamine (DA) receptor and high affinity for the 5-HT2 receptor, producing a high 5-HT2/D-2 ratio, best distinguishes atypical antipsychotics like clozapine from typical antipsychotic drugs. Through its weak antagonist action on D-2DA receptors and a potent inhibitory effect on 5-HT2 receptors, as well as its ability to increase DA and 5-HT2 release, clozapine may be able to permit more normal dopaminergic function in the anterior pituitary, the mesostriatal, mesolimbic and mesocortical regions. The numerous advantages of clozapine over typical neuroleptics are consistent with the primary importance of DA to the pathophysiology of schizophrenia. The secondary but still significant role of 5 HT in the action of clozapine may either be direct or via the effect of 5-HT on dopaminergic mechanisms. Some aspects of schizophrenia could be due to a dysregulation of the interaction between serotonergic and dopaminergic neurotransmission. PMID- 2569979 TI - Expression of (+)-3-PPP binding sites in the PC12 pheochromocytoma cell line. AB - Phencyclidine binds with high affinity to both PCP and sigma receptors. We investigated whether the clonal cell line PC12 expressed either of these receptors, and found that these cells contain a haloperidol-sensitive (+)-[3H]3 PPP binding site with a KD of 56 nM, but no PCP binding sites. The (+)3-PPP binding sites in PC12 cells displayed a reversed stereoselectivity for the benzomorphan opiates compared to CNS sigma receptors. Neither nerve growth factor nor sodium butyrate treatment affected the expression of either (+)-3-PPP or TCP binding sites in PC12 cells. PMID- 2569978 TI - Azelastine effects on electrical and mechanical activities of guinea pig papillary muscles. AB - The effects of azelastine, a new anti-asthmatic drug under clinical investigation, were studied on both normal fast action potentials (APs) and slow APs using conventional microelectrode techniques in guinea pig papillary muscles (superfused with oxygenated Tyrode solution at 37 degrees C). Slow APs were induced by either 10(-7) M isoproterenol, 10(-5) M histamine, db-cAMP (3 mM) or 10 mM TEA, in the presence of 25 mM [K]o to voltage inactive the fast Na+ channels. At 10(-5) M, azelastine depressed the maximum rate of rise (+Vmax) of the slow APs and the force of contraction. At 3 X 10(-5) M, azelastine further reduced +Vmax and the amplitude of the slow APs; complete abolishing of slow APs and contractions occurred at 10(-4) M. Upon washout of the drug, automaticity appeared. In the presence of 10(-4) M azelastine, increasing the [Ca]o concentration from 1.8 to 3.6 and 5.4 mM caused partial recovery of the slow APs and contractions. The fast APs were also depressed by azelastine. At 10(-5) and 3 X 10(-5) M, azelastine reduced +Vmax and the AP duration at 50% repolarization (APD50) of the fast APs. Complete block of the fast APs and suppression of contractions were observed after 30 min at 10(-4) M azelastine. After 3-5 h of washout, excitability recovered; however, +Vmax was depressed and APD90 was prolonged. It is concluded that azelastine inhibits the slow Ca2+ channels and the fast Na+ channels. The slow recovery suggests that the drug may accumulate inside the cells, and exert a prolonged inhibitory effect on contraction. PMID- 2569980 TI - Ifenprodil and SL 82.0715 are antagonists at the polyamine site of the N-methyl-D aspartate (NMDA) receptor. PMID- 2569981 TI - Clonidine modulates dopamine cell firing in rat ventral tegmental area. AB - The effect of clonidine (5-20 micrograms/kg i.v.) on the activity of single, identified dopamine neurons in the ventral tegmental area of the mesencephalon was studied in chloral hydrate-anesthetized male rats. Clonidine regularized cell firing without affecting the firing rate of the neurons. This effect was blocked by idazoxan (0.5 mg/kg i.v.) or yohimbine (1.0 mg/kg i.v.), but not by phentolamine (1.0 mg/kg i.v.), indicating that clonidine acts at central alpha 2 adrenoceptors. Idazoxan or yohimbine alone produced deregularization and excitation of cell firing. Pretreatment with reserpine (5 mg/kg s.c.) 4 h before the experiment abolished the neuromodulatory effect of clonidine. Thus, the regularization of ventral tegmental area dopamine cell firing by clonidine is indirect and dependent on endogenous monoamines in brain, and, in principle, a tonic adrenergic control of DA cell firing pattern is indicated. The regularization of DA cell activity produced by clonidine may underlie certain therapeutic neuropsychiatric actions of the drug. PMID- 2569982 TI - Protective action of YM-12617, an alpha 1-adrenoceptor antagonist, on the hypoxic and reoxygenated myocardium. AB - The present study was designed to determine whether the 1-form of YM-12617, which was developed recently as an alpha 1-adrenoceptor blocker, is capable of protecting the myocardium from hypoxia-induced disturbances of cardiac function and metabolism. Isolated rabbit hearts were perfused for 25 min under hypoxic conditions in the absence or the presence of 28 microM YM-12617, followed by 45 min with oxygenated perfusion medium, and functional and metabolic changes of the heart were examined. Hypoxia induced several pathophysiological changes. Upon subsequent reoxygenation, there was less than 10% recovery of the contractile force and an approximately 40% recovery of the myocardial high-energy phosphates. Treatment with YM-12617 during the hypoxic periods resulted in approximately 90% recovery of the cardiac contractile function upon subsequent reoxygenation. Treatment with YM-12617 restored the myocardial high-energy phosphates, such as ATP and creatine phosphate, to approximately 90 and 80% of the initial value, respectively, during the subsequent reoxygenation. These results suggest that YM 12617 is capable of protecting the myocardium from hypoxia-induced disturbances of cardiac function and metabolism. PMID- 2569983 TI - Neonatal W-mutant mice are favorable hosts for tracking development of marked hematopoietic stem cells. AB - Neonatal unirradiated mice of W-mutant genotypes, with a hematopoietic stem cell defect and anemia, were injected i.v. with normal fetal liver hematopoietic cells. Efficient, long-term engraftment occurred as a result of the competitive advantage to the donor stem cells. The frequency of engraftment and rate of repopulation characteristically diminish in the series W/Wv, Wf/Wf, and Wv/+, in which the severity of the endogenous defect is progressively less. H-2 compatibility is required in the inbred strain combinations examined; other histocompatibility loci play a minor role in some strain combinations. Engraftment is due to self-renewing hematopoietic stem cells ancestral to myeloid and lymphoid lineages. The more mildly defective mutants display much greater variability in the kinetics of repopulation--a result consistent with seeding by single, or very few, stem cells that form developing clones. Engraftment efficiency is reduced by prolonged culture of fetal liver cells during experimental infection by recombinant retroviruses; nevertheless, after 24 h in vitro to achieve retroviral marking, stem cells retain their ability to repopulate and develop in W/Wv neonates. PMID- 2569984 TI - Glutamate receptor activation in cultured cerebellar granule cells increases cytosolic free Ca2+ by mobilization of cellular Ca2+ and activation of Ca2+ influx. AB - The Ca2+ sensitive fluorescent probe, fura-2 has been used to monitor cytosolic free calcium levels in mature primary cultures of cerebellar granule cells during exposure to L-glutamate and other excitatory amino acids: quisqualate (QA) kainate (KA) and N-methyl-D-aspartate (NMDA). Glutamate at micromolar concentrations produced a prompt and dose-related increase in the intracellular concentration of free Ca2+, ([Ca2+]i), whereas QA, KA and NMDA had no effect. This increase was also seen in the absence of extracellular Ca2+, suggesting that L-glutamate promotes mobilization of Ca2+ from intracellular stores. In the presence of extracellular calcium, the elevation of [Ca2+]i was, in part, mediated by an increase in the plasma membrane permeability to Ca2+. This Ca2+ influx was not affected by the Ca2+-channel antagonist l-Verapamil. However, L Verapamil did block the increase in [Ca2+]i seen after depolarization of the cells with potassium. The Ca2+ response elicited by glutamate was partially blocked by the excitatory amino acid antagonist glutamate diethyl ester (GDEE). Furthermore, glutamate stimulated the formation of inositol mono-, bis-, tris- and tetrakisphosphates (IP1, IP2, IP3, and IP4) suggesting a role for these compounds for the increase in [Ca2+]i. PMID- 2569985 TI - Survival and function of aggregate cultures of rat fetal dopamine neurons grafted in a rat model of Parkinson's disease. AB - The ability to maintain tissue in culture prior to grafting would greatly facilitate the widespread application of graft therapy to neurological diseases such as Parkinson's disease. However, neurons cultured on planar substrata can be easily damaged when they are removed from the substrata and redissociated for use in grafting procedures. To overcome this limitation we utilized aggregate tissue culture methods, which allowed dopamine (DA)-rich neuronal tissue to be grafted directly following culture, without an additional redissociation. Fetal rat dopamine-neuron-containing ventral mesencephalon was cultured for 9 days in rotating flasks. The cells formed many small spheres (280 microns mean diameter), each estimated to contain about 10,000 cells. Forty such aggregate spheres were injected via a 22G needle into the DA-denervated striata of host Parkinsonian rats. A significant reduction of amphetamine-induced rotation was seen onward from 6 weeks post-transplantation, with a complete reversal of rotational asymmetry by 15 weeks post-transplantation. Well placed, surviving grafts were found in all behaviorally compensated rats (N = 6). Grafts contained an average of 517 tyrosine hydroxylase (TH)-positive neurons, as well as TH-positive fibers seen extending into the host striatum. These results suggest that aggregate culture methods are a promising means to maintain and deliver tissue for transplant therapy. PMID- 2569987 TI - Dopamine released from mesencephalic transplants restores modulation of striatal acetylcholine release after neonatal 6-hydroxydopamine: an in vitro analysis. AB - After chemical lesions which destroy the nigrostriatal dopamine pathway, transplants rich in dopamine neurons innervate the striatum and, with appropriate stimulation, drive host motor behaviors normally mediated by dopamine. We wished to determine whether dopamine released from the transplant also reinstated dopaminergic inhibition of striatal acetylcholine release. Three-day-old rat pups received bilateral intraventricular injections of 6-hydroxydopamine. Three days later cell suspensions prepared form embryonic ventral mesencephalon were injected unilaterally into the striatum. Tail pinch and amphetamine were able to elicit contralateral turning in many of these animals. Only those animals which rotated greater than or equal to 5 turns/min were included for further analysis. Subsequent assays indicated that 6-hydroxydopamine had depleted striatal dopamine to 4% of control and that the transplant had increased dopamine levels to 11% of control. Superfused striatal slices were stimulated (8 Hz, 1 min) and then exposed to amphetamine (10 microM, 3 min). The slice released dopamine, as measured by HPLC, and acetylcholine, as measured by tritium efflux after preincubation with [3H]choline. Moreover, the release of acetylcholine was inhibited by endogenous dopamine as indicated by the ability of sulpiride (1 microM) to increase tritium efflux. Striatal slices prepared from lesioned animals showed a reduction in dopamine overflow in response to both electrical stimulation (0.6% of control) and amphetamine (1% of control), and a decrease in the ability of sulpiride to increase electrically evoked acetylcholine overflow (12% of control). Transplantation partially restored the dopaminergic response to electrical stimulation (21% of control), and amphetamine (15% of control) and fully restored the sulpiride-induced increase in acetylcholine overflow (98% of control).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2569988 TI - [The regulatory effect of VIP on exploratory behavior of the rat and on the neuromediator level in the normal brain and in frontal lobectomy]. AB - In chronic experiments on 35 rats, effects on the animals behaviour of intracerebroventricular administration of vasoactive intestinal peptide (VIP) and VIP antiserum were studied under the "open field" conditions before, and on, the 4th and 9th day after local (50%) extirpation of frontal cortex. Sham-operated animals have been used as a control. A considerable regulatory action of VIP on rats' research behaviour was shown (quantity of crossed quadrants, vertical rearing) on the 9th day after the lobectomy. After the operation, the content of noradrenaline (NA), dopamine (DA) and serotonin (5HT) was determined spectrofluorometrically in cortex and subcortical structures. The VIP seemed to exert a regulatory normalizing effect on the neurotransmitters balance in cerebral structures under frontal lobectomy conditions. The most considerable effects were observed in subcortical brain structures and were due to VIP antiserum administration. PMID- 2569989 TI - [Thymus hormones--precursors of new psychoneuromodulatory peptides]. AB - The influence of thymic hormone's short fragments: thymopentin or T-5 (RKDVY), RKD and SKD, on behaviour and neurochemical processes (contents of GABA and monoamines in the brain and adrenals, plasma corticosterone), was investigated. The 100 and 500 micrograms/kg doses i.p. prevented alterations in rat's brain GABA content and plasma corticosterone level, as well as elicited anxiolytic activity in a conflict test. T-5 exerted antidepressant activity, too. SKD potentiated the haloperidol-induced catalepsy. The data obtained suggest a multicomponent (CNS-activating and inhibiting) mechanism of action of the thymic hormone fragments. These fragments seem to be formed endogenously during thymic hormone bioprocessing and may act as regulatory neuropeptides. PMID- 2569986 TI - Neurotransmitters, neuropeptides and binding sites in the rat mediobasal hypothalamus: effects of monosodium glutamate (MSG) lesions. AB - Indirect immunofluorescence histochemistry and receptor autoradiography were used to study the localization of transmitter-/peptide-containing neurons and peptide binding sites in the mediobasal hypothalamus in normal rats and in rats treated neonatally with repeated doses of the neurotoxin monosodium-glutamate (MSG). In the arcuate nucleus, the results showed a virtually complete loss of cell bodies containing immunoreactivity for growth hormone-releasing factor (GRF), galanin (GAL), dynorphin (DYN), enkephalin (ENK), corticotropin-like intermediate peptide (CLIP), neuropeptide Y (NPY), and neuropeptide K (NPK). Tyrosine hydroxylase(TH) glutamic acid decarboxylase(GAD)-, neurotensin(NT)- and somatostatin(SOM) immunoreactive (IR) cells were, however, always detected in the ventrally dislocated, dorsomedial division of the arcuate nucleus. In the median eminence, marked decreases in numbers of GAD-, NT-, GAL-, GRF-, DYN-, and ENK-IR fibers were observed. The numbers of TH-, SOM- and NPY-IR fibers were in contrast not or only affected to a very small extent, as revealed with the immunofluorescence technique. Biochemical analysis showed a tendency for MSG to reduce dopamine levels in the median eminence of female rats, whereas no effect was observed in male rats. Autoradiographic studies showed high to moderate NT binding sites, including strong binding over presumably dorsomedial dopamine cells. In MSG treated rats, there was a marked reduction in GAL binding in the ventromedial nucleus. The findings implicate that most neurons in the ventrolateral and ventromedial arcuate nucleus are sensitive to the toxic effects of MSG, whereas a subpopulation of cells in the dorsomedial division of the arcuate nucleus, including dopamine neurons, are not susceptible to MSG-neurotoxicity. The results indicate, moreover that the very dense TH-IR fiber network in the median eminence predominantly arises from the dorsomedial TH-IR arcuate cells, whereas the GAD-, NT-, GAL-, GRF- and DYN-IR fibers in the median eminence to a large extent arise from the ventrolateral arcuate nucleus. Some ENK- and NPK-positive cells in the arcuate nucleus seem to project to the lateral palisade zone of the median eminence, but most of the ENK-IR fibers in the median eminence, located in the medial palisade zone, seem to primarily originate from an area(s) located outside the arcuate nucleus, presumably the paraventricular nucleus.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2569991 TI - Somatomedin, somatostatin and psoriasis. PMID- 2569992 TI - The recombinant DNA revolution: implications for diagnosis and prevention of inherited disease. AB - Since the introduction of DNA probes as diagnostic tags, closely linked markers have been detected for 18 of the 22 most common Mendelian defects which account for 75% of the morbidity and mortality in this group. Still, as tools for diagnosis and prevention, linked probes are not completely reliable because of occasional crossovers; moreover, this diagnostic approach is not possible for sporadic cases. Fortunately, some of these limitations can be overcome by using a new strategy, termed 'reverse genetics', which aims at the detection of the primary gene defect at the DNA level and should provide the clue to the elucidation of the corresponding biochemical defect which is mostly not understood. These studies should also improve the prospects for therapy, either conventional or through replacement of the defective gene. Furthermore, new techniques have been developed in the mouse that allow the detection of specific gene defects in preimplantation embryos. If combined with in vitro fertilization, similar techniques may provide an alternative to prenatal diagnosis in situations where therapeutic abortion is not considered acceptable. PMID- 2569990 TI - Proliferation antigens in cutaneous melanocytic tumors--an immunohistochemical study comparing the transferrin receptor and the Ki 67 antigen. AB - The cellular reactivities with the monoclonal antibodies OKT9 and Ki 67 have been demonstrated to be closely related to proliferation in various malignant neoplasms. In this study a total of 25 melanocytic skin tumors was examined immunohistochemically with both antibodies and the results were evaluated semiquantitatively for OKT9 and quantitatively for Ki 67 by stereological methods. All cases of primary and metastatic malignant melanoma expressed a strong stainability for OKT9, whereas benign melanocytic nevi were almost completely negative. Our results with the monoclonal antibody Ki 67 revealed highly significant differences in the numerical density of Ki-67-positive cells between metastatic malignant melanoma (number of positive cells: 47.0 +/- 9.2 X 10(3)/mm3), primary malignant melanoma (6.3 +/- 1.9 X 10(3)/mm3) and benign melanocytic nevi (2.2 +/- 0.7 X 10(3)/mm3). Correlation analysis between mean percentage of OKT9-positive cells and numerical density of Ki-67-positive cells revealed a significant correlation of both parameters (r = 0.58; p less than or equal to 0.05), indicating a positive relationship of OKT9 and Ki 67 expression. Especially in primary malignant melanoma, however, the amount of OKT9-positive cells considerably exceeds that of Ki-67-positive cells. The monoclonal antibodies OKT9 and Ki 67 reflect 'proliferative activity' in melanocytic skin tumors, as both are expressed in significantly higher amounts in primary and metastatic malignant melanomas. The combined application of these antibodies in cutaneous melanocytic lesions might be of diagnostic and prognostic value. PMID- 2569993 TI - Azelastine in the prophylactic treatment of bronchial asthma: an Italian multicentre comparison with ketotifen. AB - The prophylactic effectiveness of the phthalazinone derivative, azelastine was compared with ketotifen. A total of 111 patients, aged 18-65 years, from 10 centres was entered into this randomized, double-blind study. All patients had reversible asthma. After 1 week on placebo, patients were allocated to either 8 mg/day azelastine once or twice daily, or to 1 mg ketotifen, twice daily, for a further 12 weeks. Azelastine was more effective in improving respiratory function (forced expiratory flow in 1 s and peak expiratory flow rate) when given in the once daily regimen, whereas clinical measures (number of weekly asthma attacks) were most improved by twice daily dosing. There was no significant difference between the effectiveness of azelastine and that of ketotifen. Treatments were equally well tolerated and a low incidence of side-effects was reported. In conclusion, 8 mg/day azelastine, in either a single or twice daily dosage regimen may be regarded as providing effective prophylaxis against bronchial asthma. PMID- 2569994 TI - Clinical experience with acrivastine--a new antihistamine. PMID- 2569995 TI - A comparison of acrivastine versus terfenadine and placebo in the treatment of chronic idiopathic urticaria. AB - Patients (n = 56) with a diagnosis of chronic idiopathic urticaria were assessed in a fully randomized, double-blind, crossover study to investigate the efficacy of acrivastine at two doses (8 and 4 mg) versus 60 mg terfenadine and placebo administered three times daily. All three active preparations were found to be effective, and significantly better than placebo, in controlling the signs and symptoms of urticaria. No significant differences were found between the active preparations, although in some cases efficacy trends favoured 8 mg acrivastine and terfenadine over 4 mg acrivastine. No significant differences were noted between the active treatments and placebo with regard to reports of drowsiness. PMID- 2569996 TI - A comparison of acrivastine versus hydroxyzine and placebo in the treatment of chronic idiopathic urticaria. AB - A total of 21 patients with a diagnosis of chronic idiopathic urticaria were assessed in a fully randomized, double-blind, crossover study to investigate the efficacy of 8 mg acrivastine versus 20 mg hydroxyzine and placebo administered three times daily. Both acrivastine and hydroxyzine were found to be effective, and significantly better than placebo, in controlling signs and symptoms of urticaria. No significant differences were found between the active preparations. Hydroxyzine was associated with significantly more reports of drowsiness than was placebo. PMID- 2569997 TI - A comparison of acrivastine versus clemastine and placebo in the treatment of patients with chronic idiopathic urticaria. AB - Twenty patients of mean age 41.3 years, with a diagnosis of chronic idiopathic urticaria were assessed in a fully randomized, double-blind, crossover study to investigate the efficacy of acrivastine at two doses (8 mg and 4 mg) versus 1 mg clemastine and placebo, given three times per day. All active preparations were found to be effective, and significantly better than placebo, in controlling the signs and symptoms of urticaria. There was a higher incidence of sedation with clemastine than with either acrivastine or placebo, although this difference did not achieve statistical significance in this small study. PMID- 2569998 TI - A comparison of acrivastine versus chlorpheniramine in the treatment of chronic idiopathic urticaria. AB - A total of 20 patients with a diagnosis of chronic idiopathic urticaria were entered into a fully randomized, double-blind, crossover study to investigate the efficacy of 8 mg acrivastine versus 4 mg chlorpheniramine three times daily. Data from 16 patients were available for analysis. Both acrivastine and chlorpheniramine were found to be effective in relieving the signs and symptoms of urticaria. There were no significant differences between the two treatments, although efficacy trends were generally in favour of acrivastine over chlorpheniramine throughout the study. PMID- 2569999 TI - Acrivastine--an evaluation of initial and peak activity in human skin. AB - Twenty-four healthy volunteers were entered into a double-blind, crossover study conducted to establish the time of onset of action and the time to peak activity of acrivastine in suppressing the weal and flare responses to intradermally injected histamine. Volunteers received single doses of 8 mg acrivastine and placebo according to a fully randomized, balanced treatment plan. Acrivastine significantly (P less than 0.001) reduced both the weal and flare responses induced by histamine challenge 30 min after oral dosing, as compared with placebo. Peak inhibition of the flare response was seen at 90 min, and maximal suppression of the weal response occurred at 120 min after administration of acrivastine. PMID- 2570000 TI - Comparison of the onset of H1-antagonism with acrivastine and terfenadine by histamine bronchial challenge in volunteers. AB - The speed of onset of H1-antagonism by 8 mg acrivastine and 60 mg terfenadine was compared by bronchial challenge of volunteers with histamine. Active treatments or placebo were administered 1 or 2 h before challenge in a double-blind, randomized, balanced crossover manner to 10 subjects. Acrivastine and terfenadine significantly attenuated the response to inhaled histamine compared to placebo. The effects of acrivastine at 1 and 2 h before challenge, and terfenadine at 2 h were indistinguishable, but that of terfenadine at 1 h was significantly less, indicating that acrivastine has a faster onset of action than terfenadine. PMID- 2570001 TI - Prolonged treatment with acrivastine for seasonal allergic rhinitis. AB - In a double-blind, placebo-controlled multicentre study, the antihistamine acrivastine, was used over prolonged periods for the treatment of seasonal allergic rhinitis. After the initial treatment period of 10 days, 8 mg acrivastine three times daily was significantly superior to placebo in controlling the symptoms of sneezing, itchy nose, running nose, watery eyes, itchy eyes and itchy throat. The benefit from acrivastine was also apparent in the second (14 days) and third (28 days) treatment periods, although the difference no longer reached statistical significance. This was probably due to the large proportion of non-responders in the placebo group who withdrew from the study owing to lack of efficacy. The investigators rated symptom control with acrivastine to be 'good' in comparison to 'poor' control with placebo treatment (P = 0.01) for all three periods. There were no significant differences between acrivastine and placebo in the incidence of adverse experiences at the end of each treatment period. Acrivastine is effective and well tolerated over prolonged periods (up to 52 days) for the treatment of seasonal allergic rhinitis. PMID- 2570003 TI - Acrivastine in allergic rhinitis: a review of clinical experience. AB - Acrivastine is an antihistamine with reduced sedating potential. This comprehensive review of clinical experience with acrivastine in allergic rhinitis considers all currently available data both published and, as yet, unpublished. Unequivocal evidence of the efficacy of 8 mg acrivastine three times daily for the control of symptoms of seasonal allergic rhinitis has been provided by 11 placebo-controlled studies involving almost 1000 patients. Additional trials have generated further supportive data as well as evidence for the use of acrivastine in the treatment of perennial allergic rhinitis. In common with most antihistamines, acrivastine alone has limited effect on the symptom of blocked nose. In a further series of 11 studies, mainly conducted in the USA, the combination of 8 mg acrivastine plus 60 mg pseudoephedrine was found to control not only the histamine-mediated symptoms of allergic rhinitis but also blocked nose. There were few adverse events associated with the use of acrivastine and the small increase in incidence of drowsiness over that found with placebo was similar to that observed for terfenadine. The marked absence of other signs of significant depression of the central nervous system (or anticholinergic activity) suggests that acrivastine will be an important addition for the antihistaminic control of symptoms of allergic rhinitis. PMID- 2570002 TI - French multicentre double-blind study to evaluate the efficacy and safety of acrivastine as compared with terfenadine in seasonal allergic rhinitis. AB - In this double-blind, multicentre study the antihistamine acrivastine was compared with terfenadine for the treatment of seasonal allergic rhinitis. The study was divided into three periods which together lasted 56 days. Patients (n = 83) were randomly assigned treatment with either 8 mg acrivastine three times daily or 60 mg terfenadine twice daily. Both agents were equally efficacious in reducing the severity of sneezing, itchy nose, blocked nose, running nose, itchy eyes, watery eyes and itchy throat as recorded daily by patients, and as rated by both the patients and their physicians at the end of each treatment period. Acrivastine and terfenadine were equally well tolerated with no serious side effects. Both effectively controlled the symptoms of seasonal allergic rhinitis in otherwise healthy individuals. PMID- 2570004 TI - Acrivastine versus terfenadine in the treatment of symptomatic dermographism--a double-blind, placebo-controlled study. AB - Twelve patients with symptomatic dermographism were entered into a double-blind, crossover study. Patients received 8 mg acrivastine three times daily, 60 mg terfenadine three times daily or placebo, according to a fully randomized balanced treatment plan. Subjective clinical assessments were performed and the response to experimentally induced dermographism was assessed. Both active treatments were well tolerated and were shown to be significantly more effective than placebo in the treatment of symptomatic dermographism and in reducing the signs and symptoms of wealing induced by a dermographometer. PMID- 2570005 TI - A multicenter, randomized, double-blind study comparing famotidine with cimetidine in the treatment of active duodenal ulcer disease. AB - The efficacy and safety of famotidine (40 mg at night), a new potent H2-receptor antagonist, has been studied in 119 patients by four investigators in four Spanish hospitals in a randomized double-blind comparative study with cimetidine (800 mg at night). Antacid tablets were allowed as additional treatment, if needed for pain relief. There were no significant differences between the groups in baseline characteristics, including duodenal ulcer size. Efficacy parameters included daytime and nocturnal symptom relief and duodenal ulcer healing, documented by endoscopy, and defined as complete reepithelization of the ulcer crater. Endoscopy was performed at baseline and after 4 and 6 weeks of treatment. One hundred and five patients fulfilled the evaluation criteria (51 patients in the famotidine group and 54 in the cimetidine group). After 4 weeks, in 91.6% of the patients receiving famotidine and 82.3% of the patients receiving cimetidine ulcers were healed. After 6 weeks, healing rates were 96% (famotidine) and 85.1% (cimetidine) (p = 0.056). Pain relief was rapid in both treatment groups, with a tendency to better response during the day in the famotidine group. The intake of antacids, as well as the clinical and laboratory safety profile were similar for both groups. PMID- 2570006 TI - Distribution of somatostatin and glucagon immunoreactive cells in the gastric mucosa of some cartilaginous fishes. AB - The comparative distribution of somatostatin- and glucagon-like-containing cells in the histomorphologically different gastric mucosae of the cartilaginous fishes Heptranchias perlo, Raja asterias, Scyliorhinus canicula, Squatina aculeata, and Torpedo marmorata was immunocytochemically studied to demonstrate a possible interrelationship between these endocrine cells and/or other endocrine or nonendocrine cells. In the gastric mucosa, these open-type glucagon and somatostatin immunoreactive cells show a double localization with different morphology and interrelationships. At the bottom of gastric pits, which corresponds to a proliferative zone, spindle or pear-shaped immunopositive cells appear rather numerously and are often in close proximity to each other. In gastric glands, triangular or oval immunopositive cells never in contact with each other were detected; their numeric ratio seems to be rather constant even if their numeric frequency and distribution vary according to the histomorphological aspect of selachian gastric glands. Glucagon immunoreactive cells seem to be more related to pepsinogenic cells, while somatostatin immunoreactive cells seem to be more ubiquitous. Both cell types can present basal cytoplasmic processes. From our results we can suggest a possible regulative role exerted by these two peptides on gastric secretion and cell proliferation. PMID- 2570008 TI - Frequent non-response to histamine H2-receptor antagonists in cirrhotics. AB - The effect of ranitidine 300 mg po given at 1800 h (famotidine 40 mg/cimetidine 800 mg) on the night time gastric pH was tested using longterm intragastric pH monitoring in 27 patients with and 32 patients without liver cirrhosis. A rise in the gastric pH above 4.0 for more than six hours between 1800 h and 0600 h was considered as sufficient effect (response) of the H2-receptor antagonists on gastric acidity. Among the patients with cirrhosis, there were significantly (p less than 0.005) more non-responders to ranitidine (16 of 27 patients) than in the control group (six of 32). When 13 of the 22 non-responders to ranitidine were subsequently treated with famotidine, only two showed a sufficient rise in their gastric pH. Of the 11 patients not responding to both H2-receptor antagonists, 10 were finally treated with cimetidine and eight did not respond. Plasma levels of all three drugs measured two and four hours after oral administration were not significantly different between cirrhotic and noncirrhotic patients as well as between responders and non-responders. In addition, in all patients plasma levels were far above the corresponding IC50 values. Therefore, differences in the absorption and plasma levels of these drugs cannot account for the frequent non-response in cirrhotics. PMID- 2570007 TI - Characterization of a Chlamydomonas transposon, Gulliver, resembling those in higher plants. AB - While pursuing a chromosomal walk through the mt+ locus of linkage group VI of Chlamydomonas reinhardtii, I encountered a 12-kb sequence that was found to be present in approximately 12 copies in the nuclear genome. Comparison of various C. reinhardtii laboratory strains provided evidence that the sequence was mobile and therefore a transposon. One of two separate natural isolates interfertile with C. reinhardtii, C. smithii (CC-1373), contained the transposon, but at completely different locations in its nuclear genome than C. reinhardtii; and a second, CC-1952 (S1-C5), lacked the transposon altogether. Genetic analysis indicated that the transposon was found at dispersed sites throughout the genome, but had a conserved structure at each location. Sequence homology between the termini was limited to an imperfect 15-bp inverted repeat. An 8-bp target site duplication was created by insertion; transposon sequences were completely removed upon excision leaving behind both copies of the target site duplication, with minor base changes. The transposon contained an internal region of unique repetitive sequence responsible for restriction fragment length heterogeneity among the various copies of the transposon. In several cases it was possible to identify which of the dozen transposons in a given strain served as the donor when a transposition event occurred. The transposon often moved into a site genetically linked to the donor, and transposition appeared to be nonreplicative. Thus the mechanism of transposition and excision of the transposon, which I have named Gulliver, resembles that of certain higher plant transposons, like the Ac transposon of maize. PMID- 2570009 TI - Magnetic resonance imaging of the hindfoot. AB - This article demonstrates normal anatomy of the foot and ankle as visualized with magnetic resonance imaging (MRI) in the sagittal, axial, and coronal planes. Additionally, selected cases chosen from our experience with more than 100 clinical scans are shown to highlight the primary areas in which we have found MRI to be clinically useful: bone marrow abnormalities, especially osteomyelitis and osteonecrosis, soft tissue injuries and masses, and cases in which metallic fixators make CT evaluation problematic. PMID- 2570010 TI - Treatment of high-pressure water gun injection injury of the foot with adjunctive hyperbaric oxygen: a case report. AB - High-pressure injection injuries are reported often in the hand and occasionally in the foot. Injection with water and air causes minimal tissue damage but nevertheless requires irrigation, minimal debridement, administration of antibiotics, and concern for development of compartment syndrome. The outcome for patients injected with water and air should be excellent. Adjunctive hyperbaric oxygen causes immediate resolution of subcutaneous emphysema, edema, and pain for more rapid rehabilitation. PMID- 2570011 TI - [Treatment of obstructive respiratory tract diseases. Principles of rational pharmacotherapy]. PMID- 2570012 TI - [Nizatidine versus ranitidine in the treatment of acute duodenal ulcer. Comparison of 300 mg nizatidine and 300 mg ranitidine in a single evening dose]. AB - In a double-blind, endoscopically controlled study on 367 duodenal ulcer patients, we compared the clinical efficacy of 300 mg ranitidine nocte with that of 300 mg nizatidine nocte, which is known to reliably provide selective inhibition of nocturnal acid secretion. Nizatidine was administered to 183, ranitidine to 184 patients. Endoscopy was performed at the start of the study, as well as at 2, 4 and 8 weeks. The presence of ulcer was defined as a benign lesion of the gastric mucosa measuring at least 5 mm in diameter; healing was characterized as complete reepithelialization. In the nizatidine group, as many as 76% of our patients were free from night pain at 2 weeks, and 88% at 4 weeks. Identical values were obtained in the group treated with 300 mg ranitidine nocte. The healing rates at 2, 4 and 8 weeks were comparable in the nizatidine and ranitidine groups (nizatidine: 57%, 87%, 92%, respectively; ranitidine: 63%, 90%, 96%, respectively). Clinically significant adverse effects were seen in neither of the two treatment groups. These results demonstrate that selective inhibition of nocturnal acid secretion achieves healing of duodenal ulcer and freedom from pain as rapidly and effectively as protracted inhibition of acid secretion provided by the administration of 300 mg ranitidine nocte. PMID- 2570013 TI - Plasma membrane lipids modulate the response to water deprivation in rat kidney. AB - The lipid composition of biological membranes determines many of the cellular responses to stimuli such as hormones and drugs. It is known that the neonate has differences in renal function compared to the adult individual, which determine the characteristics of pharmacokinetics and pharmacodynamics at this age. We studied the lipid composition of renal plasma membranes, the renal response to water deprivation, and the activity of plasma membrane gamma-glutamyl transpeptidase (GGTP) in newborn, 21-d-old, and adult rat. It was found that the cholesterol/phospholipid (CH/PL) ratio of newborn membranes was significantly higher than that of membranes from 21-d-old and adult rats, which were similar. These findings paralleled low ability to concentrate urine in the newborn and higher sensitivity of the neonatal kidney to water deprivation. Kidney membranes from 21-d-old and adult rats showed similar lipid composition, and the response to water deprivation was also similar. The changes observed in GGTP activity at different ages varied with the cholesterol content of kidney plasma membranes. These findings suggest participation of the lipid composition of kidney membranes in modulation of the renal response to water deprivation. PMID- 2570014 TI - [An experimental study of spontaneously diabetic Bio Breeding/Worcester (BB/W) rats--with special references to the rat MHC (RT1) in the development of the disease]. AB - The role of rat MHC (RT1) in the development of diabetes mellitus was studied on the Bio Breeding/Worcester (BB/W) rat, an experimental model animal for human juvenile onset (Insulin-dependent, Type 1) diabetes mellitus. The rate of the development of diabetes mellitus was as follows; 35/45 (77.8%) in inbred BB/W rats (u/u haplotype), and 5/397 (1.7%) in F2 rats. Histopathological examinations demonstrated that there were lymphocytic thyroiditis, T cell depletion in the spleen and in the lymph nodes, and extramedullary hematopoiesis, in addition to typical lymphocytic insulitis in the pancreas. Immunohistochemically, the expression of class II antigens in the pancreas was shown to be highly concerned with the histopathological changes and the clinical onset of the diabetes mellitus. Analysis of subsets of infiltrating cells demonstrated that macrophages appeared in the pancreas prior to the onset of the disease. And it was suggested that the infiltrated OX 6+ macrophages functioned as antigen presenting cells, leading to expression of the class II antigens on the surface of islet cells, and finally caused severe lymphocytic insulitis. However, BB/W rats showed complete negative immunohistochemical stainings with a monoclonal antibody HOK 2B, which stained other rat strains carrying RT1u class II antigen (TO, SDJ). Moreover, differences in blocking effect of HOK 2B were found on mixed lymphocyte reactions (MLR) between BB/W and other strains with RT1u haplotype. These findings raise the possibility that minor heterogeneities exist in the structure of class II molecules in RT1u rats including BB/W, which might be closely related to the onset of the disease. PMID- 2570015 TI - Cystic fibrosis in Finland: a molecular and genealogical study. AB - The incidence of cystic fibrosis (CF) in Finland is one tenth that in other Caucasian populations. To study the genetics of CF in Finland, we used a combined molecular and genealogical approach. Out of the 20 Finnish families with a living CF patient, 19 were typed for eight closely linked restriction fragment length polymorphisms (RFLP) at the MET, D7S8, and D7S23 loci. The birthplaces of the parents and grandparents were traced using population registries. Allele and haplotype frequencies in Finland are similar to those of other European and North American populations, but are modified by sampling: two regional CF gene clusters, evidently the results of a founder effect, were identified. Generally, the gene was evenly distributed over the population, carrier frequency being estimated at approximately 1.3%. We conclude that CF in Finland is caused by the common Caucasian mutation(s), and that the low frequency of the gene can be explained by a negative sampling effect and genetic drift. PMID- 2570016 TI - BamHI and SacI RFLPs of the human immunoglobulin IGHG genes with reference to the Gm polymorphism in African people. Evidence for a major polymorphism. AB - In this paper, we extend the study of the IGHG gene RFLPs in black African persons and in some other individuals characterized by a Negroid admixture. We demonstrate a polymorphism that is much more important in black Africans, than in Caucasoids, mainly for the IGHG3 and G1 genes, the most 5' members of the IGHG multigene family. These genes encode for the IgG3 and IgG1 subclasses, which are of crucial biological importance. PMID- 2570017 TI - Risks of fetal cystic fibrosis based on linkage disequilibrium data. AB - First-trimester prenatal diagnosis of cystic fibrosis depends on tissues being available from a previous affected child for determination of the phase relationship of DNA markers. If no such tissues are available, it is possible to estimate the risks of a couple producing an affected child from the distribution of haplotypes showing linkage disequilibrium with the cystic fibrosis gene. We have calculated all the fetal risk subsets from the various parental haplotype combinations for the restriction fragment length polymorphisms identified by the KM.19/PstI and XV-2c/TaqI systems. We conclude that only in a limited number of parental combinations are the fetal risks sufficiently high or sufficiently low to be used in prenatal diagnosis. PMID- 2570018 TI - Selection of human chromosome 21-specific DNA probes for genetic analysis in Alzheimer's dementia and Down syndrome. AB - We used a mouse-human somatic cell hybrid to construct a chromosome 21-enriched library in phage vector EMBL4. In all, 35 phage clones containing human inserts were identified by differential screening with total human and mouse DNA. Whole recombinant phages were regionally mapped on chromosome 21 by Southern blot analysis using competitive hybridisation conditions to block repetitive sequences. Ten phage clones mapped proximal to a translocation breakpoint in band 21q21.2, while 25 mapped distal to this point. Three of the phage clones identify restriction fragment length polymorphisms. Polymorphic chromosome 21 markers may be useful in the genetic analysis of Alzheimer's dementia and Down syndrome. PMID- 2570019 TI - Assignment of anonymous DNA probes to specific intervals of human chromosomes 16 and X. AB - Anonymous DNA probes mapping to human chromosome 16 and the distal region of the human X chromosome were isolated from a genomic library constructed using lambda EMBL3 and DNA from a mouse/human hybrid. The hybrid cell contained a der(16)t(X;16)(q26;q24) as the only human chromosome. Fifty clones were isolated using total human DNA as a hybridisation probe. Forty six clones contained single copy DNA in addition to the repetitive DNA. Pre-reassociation with sonicated human DNA was used to map these clones by a combination of Southern blot analysis of a hybrid cell panel containing fragments of chromosomes 16 and X and in situ hybridisation. One clone mapped to 16pter----16p13.11, one clone to 16p13.3--- 16p13.11, four clones to 16p13.3----16p13.13, two clones to 16p13.13----16p13.11, one clone to 16p13.11, seven clones to 16p13.11----16q12 or 16q13, four clones to 16q12 or 16q13, three clones to 16q13----16q22.1, four clones to 16q22.105--- 16q24, and nineteen clones to Xq26----Xqter. Two clones mapping to 16p13 detected RFLPs. VK5 (D16S94) detected an MspI RFLP, PIC 0.37. VK20 (D16S96) detected a TaqI RFLP, PIC 0.37 and two MspI RFLPs, PIC 0.30 and 0.50. The adult polycystic kidney disease locus (PKD1) has also been assigned to 16p13. The RFLPs described will be of use for genetic counselling and in the isolation of the PKD1 gene. Similarly, the X clones may be used to isolate RFLPs for genetic counselling and the isolation of genes for the many diseases that map to Xq26----qter. PMID- 2570020 TI - Multiple mutations underlying familial hypercholesterolemia in the South African population. AB - Ten restriction fragment length polymorphisms of the LDL receptor gene were used for haplotype analysis in 12 unrelated patients with homozygous familial hypercholesterolemia. These patients were drawn from the Black, Coloured, and White population groups and collectively represent 24 mutant alleles underlying the FH phenotype. Five distinct haplotypes were detected. Hybridization analysis using DNA codigested with EcoRI and PstI revealed that haplotype IV was associated with two distinct mutations. When coupled to the recent demonstration by other workers of two receptor defects in South African Afrikaners homozygous for FH and haplotype I, these data are suggestive of at least seven distinct LDL receptor mutations in the FH patients examined and thus in the general South African population. PMID- 2570021 TI - Long-range restriction map of a region of human chromosome 19 containing the apolipoprotein genes, a CLL-associated translocation breakpoint, and two polymorphic MluI sites. AB - The apolipoprotein gene cluster on human chromosome 19 (APOC1, APOC2, APOE) has been localised by pulsed-field gel electrophoresis to within 200 kb of a chronic lymphocytic leukemia-associated translocation breakpoint. A restriction map covering 1300 kb around these loci has been constructed and contains two polymorphic MluI sites, which appear to show Mendelian inheritance. The orientation of the map on the chromosome has been established as 19cen - CLL breakpoint - APOC2 - 19qter. Pedigree analysis using APOC2, a probe derived from the CLL breakpoint, and other localised markers on 19q suggests that the myotonic dystrophy locus is distal to APOC2 on 19q. PMID- 2570022 TI - Detection of an unbalanced translocation (4;14) in a mildly retarded father and son by flow cytometry. AB - A child with impaired intelligence, minor dysmorphisms, obesity and genital hypoplasia was found to have an apparently balanced translocation, 46,XY,t(4;14)(q12;q13), following cytogenetic analysis. The same rearrangement was also detected in the child's father, who had similar phenotypic abnormalities to his son. Detailed study of flow karyotypes produced from lymphoblastoid cell lines established that in both patients the translocation was in fact unbalanced with approximately 11 million base pairs of DNA (corresponding to about 6.0% of chromosome 4 or 11.0% of chromosome 14) being lost. PMID- 2570023 TI - Molecular analysis of 46,XY females and regional assignment of a new Y-chromosome specific probe. AB - The relationship between Y-chromosome abnormalities and gonadal differentiation was investigated in six phenotypic females with a 46,XY karyotype and one patient with ambiguous genitalia secondary to apparently nonmosaic 46,XY mixed gonadal dysgenesis. No alterations were found in the Y chromosomes of six of these individuals by the use of either cytogenetic or molecular techniques. Cytogenetic analysis with high-resolution G-banding and Q-banding revealed a small deletion in the short arm of the Y chromosome in one female patient with some features of Turner syndrome. Southern hybridization with Y-specific probes showed a loss of DNA within deletion intervals 1, 2, and 3 of the Y chromosome. A new Y-chromosome specific DNA probe that hybridizes to deletion interval 3 is described. PMID- 2570025 TI - Isolation and characterization of a human variable copy number tandem repeat at Xcen-p11.22. AB - A subclone (M27B) has been isolated from a cosmid randomly selected from a library enriched for human X-chromosomal material. The subclone is extensively single-copy sequence, but also contains three complete copies and one partial copy of a 26-bp repeat, within which exists an inverted repeat having the potential to form a cruciform loop structure. Genomic sequences in this repeat region are apparently refractory to cloning, and rearrangements occurring during this process result in deletions. M27B detects multiple X-linked restriction fragments for a wide range of enzymes including MspI and HpaII. We have assigned the locus recognized by the probe (DXS255) to Xcen-Xp11.4 by mapping with a somatic cell hybrid panel and further refined its localization to Xp11.22 by in situ hybridization. The repeat sequence that is presumed to be responsible for the hypervariability observed does not show close similarity to other variable copy number tandem repeats described. PMID- 2570024 TI - The Mos proto-oncogene maps near the centromere on mouse chromosome 4. AB - The Mos proto-oncogene, the cellular homolog of the transforming gene of Moloney murine sarcoma virus, was originally assigned to mouse chromosome 4 using independent panels of mouse/hamster somatic cell hybrids. By in situ hybridization to metaphase chromosomes and standard genetic backcrosses, we have confirmed this assignment and determined that Mos maps near the centromere in a region devoid of other markers. We have also identified a restriction fragment length polymorphism (RFLP) that defines two alleles of the Mos locus in selected inbred strains of laboratory mice. Using the RFLP, we determined the strain distribution pattern for the Mos gene in three sets of recombinant inbred strains and in five strains congenic for histocompatibility antigen genes localized on chromosome 4. These results establish Mos as a useful marker in a poorly characterized region of the mouse genome. In addition, these results will facilitate the genetic analysis of the Mos locus. PMID- 2570027 TI - The cardiac actin locus (Actc-1) is not on mouse chromosome 17 but is linked to beta 2-microglobulin on chromosome 2. AB - A restriction fragment variant and recombinant inbred strains were used to show that the cardiac actin locus (Actc-1) is closely linked to beta 2-microglobulin (B2m) and several other loci on chromosome 2 of the mouse. Close linkage of Actc 1 and B2m in both man and mouse provides another example of a chromosomal segment that has been conserved since the divergence of the lineages leading to these two species. PMID- 2570026 TI - Protein kinase C: a new linkage marker for growth hormone and for COL1A1. AB - An expanded linkage group on the long arm of human chromosome 17 is reported. Using the CEPH panel of DNAs and restriction fragment length polymorphism (RFLP) markers for the centromere locus (D17Z1), growth hormone (GH1), collagen type I alpha 1 (COL1A1), and protein kinase C-alpha polypeptide (PKCA) loci, theta values of 0.03, 0.11, and 0.23 were found between PKCA and GH1, PKCA and COL1A1, and PKCA and D17Z1, respectively. The theta values calculated for GH1 versus COL1A1 or D17Z1 were 0.11 and 0.23, respectively. Sex-specific recombination rates were calculated for the best likelihood order and demonstrate female recombination greater than male recombination. Therefore, the loci studied span a map region of approximately 30 cm between 17cen and 17q24, with the most likely gene order being D17Z1-COL1A1-PKCA-GH1. PMID- 2570028 TI - Comparison of linkage maps of mouse chromosome 12 derived from laboratory strain intraspecific and Mus spretus interspecific backcrosses. AB - Progeny from an interspecific backcross between laboratory mice and Mus spretus were typed for inheritance of eight genetic markers on chromosome 12. Marker order determined by segregation analyses of 115 meiotic events was in good agreement with that determined previously using intraspecific laboratory strain backcrosses. Two additional markers, D12Nyu5 and Lamb-1, previously not ordered, were located in the middle of the interval between D12Nyu12 and D12Nyu1. Marker spacing was reduced in the interspecific cross relative to that observed in intraspecific crosses. Furthermore, the interspecific cross was characterized by marked deviation from 1:1 segregation in the recombinant chromosomes and very strong positive interference. These data suggest that comparisons of different mouse crosses may facilitate the understanding of underlying mechanisms that govern recombination events in complex genomes. PMID- 2570029 TI - A physical map around the WAGR complex on the short arm of chromosome 11. AB - A long-range restriction map of part of the short arm of chromosome 11 including the WAGR region has been constructed using pulsed-field gel electrophoresis and a number of infrequently cutting restriction enzymes. A total of 15.4 Mbp has been mapped in detail, extending from proximal 11p14 to the distal part of 11p12. The map localizes 35 different DNA probes and reveals at least nine areas with features characteristic of HTF islands, some of which may be candidates for the different loci underlying the phenotype of the WAGR syndrome. This map will furthermore allow screening of DNA from individuals with WAGR-related phenotypes and from Wilms tumors for associated chromosomal rearrangements. PMID- 2570030 TI - Localization of the properdin structural locus to Xp11.23-Xp21.1. AB - Properdin is a serum protein belonging to the alternative pathway of complement activation whose absence is often associated with fatal bacterial infections. Properdin deficiency segregates with an X-linked recessive pattern and its position has been recently refined by genetic linkage analysis to the proximal part of the X-chromosome short arm near the OTC and DXS7 loci. We have hybridized an 0.8-kb genomic clone encoding part of the human properdin gene to a panel of somatic cell hybrids retaining different portions of the human X chromosome and thereby localized the probe to Xcen-Xp21.1. Furthermore, in situ hybridization of the same probe to replication banded metaphase chromosomes refined this localization to the region Xp11.23-Xp21.1 (with a peak grain distribution in the region equivalent to Xp11.4). As OTC and DXS7 map to Xp21.1 and Xp11.3, respectively, the data presented here strongly suggest that the X-linked deficiency syndrome is due to a defect in the locus encoding the structural properdin gene or in a physically close regulatory locus. PMID- 2570032 TI - Isolation and regional localization of the murine homeobox-containing gene Hox 3.3 to mouse chromosome region 15E. AB - A murine homeobox-containing cDNA clone has been isolated from an adult spinal cord library. Using in situ hybridization and somatic cell genetics techniques, the newly isolated homeobox gene has been mapped to mouse chromosome region 15E. Because of its chromosomal location, we called this gene locus Hox-3.3. Nucleotide sequence analysis revealed that the Hox-3.3 gene represents the murine cognate of the human homeobox gene c8. The presumptive organization of the murine Hox-3 homeobox gene cluster is discussed. PMID- 2570031 TI - The murine retinoblastoma homolog maps to chromosome 14 near Es-10. AB - Restriction fragment length variants have been exploited to map genetically Rb-1, the murine homolog of the human retinoblastoma gene. Rb-1 localized to mouse chromosome 14 on the basis of results from analysis of somatic cell hybrids. In an interspecific backcross involving Mus spretus, Rb-1 and the murine homolog of the human esterase D gene (ESD), which we refer to here as Esd, were inseparable. Furthermore, the strain distribution patterns of Rb-1 and Es-10 are the same in 31 of 32 recombinant inbred strains. Close linkage of the chromosome 14 morphological marker hairless (hr) to Rb-1 is also implied. These results localize Rb-1 on the mouse linkage map and provide close genetic markers to follow Rb-1 in somatic as well as in germline genetic experiments. Additionally, the results suggest that Es-10 is the murine homolog of ESD and provide further evidence for linkage conservation during mammalian evolution. PMID- 2570034 TI - CD4 expression and function in HLA class II-specific T cells. AB - The CD4 molecule is currently the object of intense interest and investigation both because of its role in normal T-cell function, and because of its role in HIV infection. Our studies of CD4 expression and function on class II-specific T cells indicate that: 1) CD4 expression is generally a marker of T cells that are committed to class II antigen recognition, even when they have cytotoxic function; 2) CD4 is typically involved in the proliferative and cytotoxic function of such cells, which we hypothesized to be due to interaction with a nonpolymorphic determinant on class II; 3) the importance of CD4 to the recognition is most prominent in T-cell interactions with low "avidity"; and 4) although CD4 may play a role in adhesion per se, the majority of its function seems to be in a post-adhesion phase of cytotoxic T-cell recognition. PMID- 2570033 TI - The long-range restriction map surrounding the mouse agouti locus reveals a disparity between physical and genetic distances. AB - Isolation of a gene based on its location, which depends on aligning physical landmarks with the genetic map, can yield basic information about genome structure and organization. As a first step toward isolating the mouse agouti (A) locus, we have begun to define the physical position of this gene relative to genetically linked DNA probes from the Psp, Emv-15, and Src loci. Using a combination of pulsed-field gel techniques that include partial digestion with rare-cutting restriction enzymes and analysis of polymorphic sites present in certain inbred strains, we have constructed long-range restriction maps for each of the probes that span a total of more than 3000 kb. The Src and Emv-15 probes are less than 600 kb apart, but are separated from the Psp probe by at least 1500 kb. By determining the position of a 75-kb deletion that inactivates agouti function, we have localized the A locus to within 500 kb of the Psp probe, but more than 600 kb away from the Emv-15 probe. These physical distances contrast with the known recombination frequencies, 3 +/- 3 cM for A-Psp and less than 0.3 cM for A-Emv-15, and suggest that recombination between A and Emv-15 may be suppressed. PMID- 2570035 TI - A role for CD4+ but not CD8+ T cells in immunity to Schistosoma mansoni induced by 20 krad-irradiated and Ro 11-3128-terminated infections. AB - The role of CD4+ (L3/T4+) and CD8+ (Lyt-2+) T cells in immunity to Schistosoma mansoni induced by 20 krad-irradiated and Ro 11-terminated infections in mice was investigated directly by in vivo depletion of these subsets with cytotoxic rat monoclonal antibodies (mAb). Effective physical depletion was demonstrated by flow cytometric analysis and immunohistochemical staining. Functional depletion of helper activity following anti-CD4 treatment was indicated by an abrogation of concanavalin A(Con A)-induced colony-stimulating factor (CSF) release, while anti CD8 treatment had no effect in these assays. Pre-existing S. mansoni-specific antibody levels were unaffected by anti-CD4 and anti-CD8 treatment. In vivo depletion of CD4+ T cells resulted in a dramatic reduction in immunity induced by one (up to 100%) and two (up to 70%) vaccinations with 20 krad-irradiated cercariae and also of resistance induced by Ro 11-attenuated infections (up to 100%). Depletion of CD8+ T cells had no effect on resistance induced by any of the vaccination protocols investigated. A correlation was observed between resistance and T cell-induced, macrophage-mediated killing of schistosomula in vitro, both of which were abrogated following anti-CD4 treatment but were unaffected by CD8+ T-cell depletion. The possible role of CD4+ T cells in vivo and the implications for vaccine development are discussed. PMID- 2570036 TI - CD11/CD18-independent neutrophil adherence to inducible endothelial-leucocyte adhesion molecules (E-LAM) in vitro. AB - We examined the mechanisms involved in neutrophil adherence to cultured human umbilical vein endothelial cells (HEC) induced by direct stimulation of the neutrophils by phorbol myristate acetate (PMA), formylmethionyl-leucyl phenylalanine (FMLP), or the calcium ionophore A23187 (neutrophil-dependent adherence), or by pretreatment of HEC with interleukin-1 (IL-1), tumour necrosis factor (TNF) or lipopolysaccharide (LPS) (endothelial-dependent adherence). Two distinct mechanisms for neutrophil adherence to HEC were demonstrated by performing adherence assays: (i) at 37 degrees versus 4 degrees; (ii) in the presence of Ca2+ only versus Mg2+ only; and (iii) in the presence or absence of monoclonal antibodies (mAb) to the CD11/CD18 adhesion complex of neutrophils. A CD11/CD18-dependent mechanism (i.e. inhibited by anti-CD18 mAb) was identified that was active in the presence of Mg2+ only but not of Ca2+ only, and at 37 degrees but not at 4 degrees. A CD11/CD18-independent mechanism (i.e. not inhibited by anti-CD18 mAb) was active at 4 degrees and at 37 degrees, and in the presence of Ca2+ only and of Mg2+ only. Neutrophil-dependent adherence induced by FMLP or PMA occurred solely via the CD11/CD18-dependent mechanism, whereas endothelial-dependent adherence induced by a 4-hr pretreatment with IL-1, TNF, or LPS involved both CD11/CD18-dependent and/independent mechanisms. CD11/CD18 deficient neutrophils isolated from a patient with leucocyte adherence deficiency (LAD) maintained the ability to adhere to LPS-pretreated HEC in the presence of Ca2+ only, indicating that this mechanism of adherence involves a receptor on the neutrophil distinct from CD11/CD18. Furthermore, the disappearance of the CD11/CD18-independent, but not of the CD11/CD18-dependent mechanism of adherence, in HEC treated with TNF for 24 hr suggests that the two mechanisms of neutrophil adherence also involve distinct inducible endothelial-leucocyte adhesion molecules (E-LAM). PMID- 2570038 TI - Circulating human T and B lymphocytes express the p55 interleukin-2 receptor molecule (TAC, CD25). AB - Using a highly sensitive immunofluorescence procedure 15-45% of blood lymphocytes from normal human donors can be shown to react with anti-TAC (CD25) monoclonal antibodies. The standard procedure for indirect immunofluorescence does not allow the detection of these CD25-positive cells. Ten healthy adults were tested, and all showed a population of positive cells. Three donors have been tested more than once, and have given consistent results. Three different CD25 antibodies were used, again with consistent results. Double-marker studies showed that the positive cells included B cells, as well as CD4-bearing and CD8-bearing T cells. PMID- 2570037 TI - Rat mesangial cells actively produce phosphatidylinositol-anchored Thy-1. AB - We provide evidence that the mesangial cells of rat kidney glomeruli express Thy 1 as a phosphatidylinositol-anchored protein. Both the mesangial area of kidney, examined in tissue sections, and mesangial cells maintained in culture for more than 3 months, showed prominent immunofluorescence staining with an anti-Thy-1 monoclonal antibody (OX7); this staining was almost completely abolished by pretreating kidney sections or mesangial cells with the phosphatidylinositol specific enzyme, phospholipase C. By Northern blotting, mesangial cells were shown to express mRNA of an appropriate size, hybridizing to a mouse Thy-1.1 specific probe. PMID- 2570039 TI - Gm allogenotype distribution and lack of HLA-DR, Gm interaction in SLE and CREST/PSS. AB - The distribution of Gm allogenotypes (Gm allotypes identified by IgCH restriction fragment length polymorphism (RFLP) analysis) was compared in 66 systemic lupus erythematosus (SLE) patients, 38 CREST/PSS group (P less than 0.05) but not in the SLE group. HLA-DR5 is significantly increased in frequency in this series of CREST/PSS patients compared to controls (P less than 0.001), and log linear regression analysis showed that although both DR5 and Gm homozygosity were significant factors determining disease susceptibility, there was no evidence of an interactive effect between these two groups of genes increasing the predisposition to CREST/PSS. PMID- 2570040 TI - Neuroimmunomodulation: facts and dilemmas. PMID- 2570041 TI - Production of IL-2 and IFN by TH2 clones. AB - We have studied the production of IL-2, IL-4 and IFN-gamma by a panel of CD4+ clones produced in our laboratory. The clones were classified as TH1 and TH2 because of their ability to secrete IL-2 or IL-4, respectively, following stimulation with APC + Ag and by their characteristic proliferative responses to exogenous IL-2 or IL-4. Some of the TH2 clones, all of which happened to be autoreactive, produced IL-2 and one of these, as well as one antigen-reactive TH2 clone, also secreted IFN-gamma following stimulation with immobilized anti-CD3 mAb. IL-2 production by TH2 cells required higher concentrations of anti-CD3 mAb than IL-4 production. Thus, the TH2 clones seem to be heterogeneous. We designate the IL-2/IL-4 secretors as TH2B and those making IL-4 as TH2A clones. PMID- 2570042 TI - Alloreactive T cell recognition of the HLA-DR beta N-terminal polymorphic region. AB - T cell alloreactivity, originally discovered as a tissue transplantation effect, is believed to be a manifestation of the normal major histocompatibility complex (MHC) restriction of antigen presentation by accessory cells to T cells. The molecular features of the human class II-MHC proteins (HLA) which are recognized by alloreactive T cells are not at present understood, although they are clearly related to the polymorphic nature of the MHC proteins. Human CD4+ T cell clones were selected by response to the HLA-DR2 peptides beta 1-15 or beta 51-65, in an MHC-restricted manner. In addition, these clones respond to cell lines expressing the DR2 haplotype, without the requirement for accessory antigen presenting cells. DR2 beta peptide 1-15 blocks the T cell alloresponse and polymorphic residues are shown to stimulate the peptide-specific response of these clones. Thus, the polymorphic residues contained within the DR beta sequence 1-15 are demonstrated to be directly recognized by alloreactive T cells. PMID- 2570043 TI - The value of the left ventricular ejection fraction in the treatment of angina pectoris following acute myocardial infarction: a randomized double blind study. AB - Radionuclide determination of left ventricular ejection fraction was performed at hospital discharge and one month later in 60 patients who had suffered acute myocardial infarction. At the first determination, the patients were randomized into two groups. In the first group, the cardiologist who cared for the patients was provided with the result of the determination of the ejection fraction whereas, in the second group, the result was withheld. At a 'blinded' evaluation two months after hospital discharge, 7 of those patients (24%) from the group where the cardiologist knew the ejection fraction and 11 of the patients (38%) in whom this result was withheld complained of angina pectoris on exertion (ns). The medication of the patients did not differ in the two groups. No significant difference was found in the values of the ejection fraction in patients with and without angina pectoris. In this controlled study, we were not able to document a clinical effect by routinely determinating left ventricular ejection fraction in patients with acute myocardial infarction in the treatment of angina pectoris. PMID- 2570044 TI - Mosquitoes: how to be the perfect host. PMID- 2570046 TI - Dynorphin deficiency in Tourette's syndrome. PMID- 2570045 TI - Demonstration of neuroendocrine cells in ovarian mucinous tumors. AB - The frequency of argyrophil cells in mucinous cystadenocarcinomas, borderline tumors (MBT) and cystadenomas was 29.8% (14 of 47), 46.7% (7 of 15) and 11.1% (2 of 18), respectively. These were statistically higher than the frequencies in 17 clear cell carcinomas, 43 serous cystadenocarcinomas, and 24 metastatic carcinomas. Immunoreactive cells for serotonin, somatostatin, gastrin, pancreatic polypeptide, growth hormone-releasing hormone, metenkephalin, neuron-specific enolase, and chromogranin-A were detected in almost all these cases with argyrophil cells. However, immunoreactivities for glucagon, vasoactive intestinal polypeptide, and adrenocorticotropic hormone were negative in ovarian mucinous tumors. Immunohistochemical multiplicity of neurohormones was remarkable in 15 MBT (including 5 mullerian and 10 intestinal MBT) and it was not related to the number of argyrophil cells per unit tumor cells. Individual hormones demonstrated here seemed to be present in different cells, but certain cells were immunoreactive for both gastrin and somatostatin by double immunostaining. Based on the high frequency of endocrine cells, borderline tumors seemed to be unique in the spectrum of mucinous ovarian tumors. PMID- 2570047 TI - c-erB-2 oncoprotein expression in malignant and nonmalignant breast tissue. AB - Using a specific polyclonal antibody (21N) to the c-erbB-2 oncoprotein, a total of 200 primary breast carcinomas of various histological types and 46 benign breast lesions, were immunohistochemically stained, to identify those tissues over-expressing this putative transmembrane receptor. Positivity for this oncoprotein was evident as an intense brown granular staining predominantly located at the cell membrane and was present in 17% (n = 34) of the primary breast carcinomas investigated. With the exception of one case of apocrine metaplasia, none of the remaining benign breast samples (n = 45) were positive for this oncoprotein. This study shows that overexpression of the c-erb B-2 oncoprotein is easily identifiable and can be detected on paraffin embedded sections using an immunohistochemical technique. The results obtained correlate well with other c-erbB2 detection systems and in addition, positivity for this oncoprotein is mostly confined to the malignant phenotype. PMID- 2570048 TI - Serotonin-like immunoreactive cells in the pulmonary epithelium of ancient fish species. AB - The pulmonary mucosa of three species of ancient fish was studied immunohistochemically to show the distribution of serotonin, regarded as the main monoamine of mammalian bronchopulmonary paraneurons. Serotonin-like immunoreactive cells, dispersed through the airway epithelium as single cells, were found in all the fish species studied. They are presumably equivalent to the neuroendocrine cells reported in the lungs of mammalian and submammalian vertebrates. However, the precise role and the function of these cells remain unknown. Since the species studied belong to the most primitive extant groups of ancient fish, the present investigation suggests that serotonin is widely distributed in the lungs of the vertebrates. Several peptides, known to be specific cytochemical markers for the identification of the pulmonary neuroendocrine cells of mammals, are being investigated in the lungs of the fish species studied. They may help to trace the phylogeny of the pulmonary neuroendocrine cell system and to elucidate its function in lower vertebrates. PMID- 2570049 TI - Activation of human CD8+ suppressor T cells by an antigen-specific CD4+ T-cell line in vitro. AB - We generated an antigen (streptococcal cell wall antigen)-specific T-cell line from peripheral blood mononuclear cells of a healthy donor with an intermediate response to streptococcal cell wall antigen. Proliferation of the T-cell line was completely blocked by a monoclonal antibody to HLA-DR. This line activated autologous CD8+ T cells in an antigen-specific manner in the presence of autologous monocytes. This activation was mediated by a factor derived from this line and was blocked by a monoclonal antibody against HLA class I molecules. The resultant CD8+ T blasts showed antigen-nonspecific suppression but no cytolytic activity. This antigen-specific generation of the CD8+ T-cell line in vitro by the antigen-specific CD4+ T-cell line is expected to contribute to analyses of functions of CD8+ T-cell subsets, particularly in the down-regulating system, at both cellular and molecular levels. PMID- 2570050 TI - RFLP-defined HLA-DP polymorphism in four ethnic groups. AB - Patterns of BglII, MspI, and TaqI DP alpha and DP beta hybridizing restriction fragment length polymorphisms are compared with primed lymphocyte typing- determined specificities in 28 10th International Histocompatibility Workshop core cell lines. Correlation of specific RFLP patterns with most recognized DP types is confirmed, although the RFLPs do not distinguish DPw3 from DPw6. Analysis of DP-region nucleotide sequence data shows that the observed RFLPs are those expected to hybridize to the DP alpha and beta probes and are thus not due to cross-hybridization to other genes. The distribution of RFLPs in Chinese, Micronesian, South Indian, and white Australian populations is described. The most frequent DP specificity in Chinese is DPw5. In the other populations DPw4 is the most common specificity, although DPw5 is also relatively common in Micronesians. Four new DP alpha RFLP patterns and three new DP beta patterns are described. There are also numerous unusual combinations of DP alpha and DP beta alleles particularly in the South Indian population. PMID- 2570051 TI - HLA-D typing of heterozygotes using restriction fragment length polymorphism in the HLA-DQ gene region on the basis of standard band patterns derived from HLA homozygotes. AB - Genomic DNAs from 37 Epstein-Barr virus-transformed HLA-D homozygous cell lines with different Dw specificities, Dw1-Dw23, and DKT2 were digested with four different restriction endonucleases (EcoRI, PstI, TaqI, and MspI) and hybridized to DQ alpha and DQ beta cDNA probes. Polymorphic patterns of multiple fragments correlating with Dw, DQ, and DR haplotypes were detected, and the restriction fragment length polymorphism standard pattern specific for each Dw, DQ, and DR specificity could be defined. The polymorphic fragment patterns of HLA-D heterozygotes were predicted simply by the summation of two standard patterns of HLA-D homozygotes and utilized to identify HLA-D specificities of 25 normal individuals, who are HLA heterozygotes in most cases. The HLA-D, -DR, and -DQ specificities defined by this DNA typing were compared with those assigned by serologic and cellular typing. There was good correlation, allowing the application of accurate genotyping by DQ alpha and DQ beta cDNA probes to HLA class II typing of HLA heterozygotes. PMID- 2570052 TI - Use of alpha 2-adrenergic receptor antagonists. PMID- 2570053 TI - Marketing the allied health professions. PMID- 2570054 TI - Effects of excitant amino acids on acoustic responses of inferior colliculus neurons. AB - Iontophoretic application of the excitant amino acids (EAAs), glutamate, aspartate and N-methyl-D-aspartate (NMDA) resulted in increased acoustically evoked and spontaneous firing of most neurons in the central nucleus of inferior colliculus (ICC). The excitatory effects of these EAAs were blocked by simultaneous application of EAA antagonists which selectively block the NMDA receptor subtype, 2-amino-5-phosphonovalerate or D-alpha-aminoadipate and to a lesser extent with non-selective EAA antagonists, such as glutamic acid diethylester. Application of NMDA receptor-selective EAA antagonists alone greatly reduced the firing of most ICC neurons examined, but non-selective EAA antagonists either increased or produced little change in firing of most ICC neurons examined. In this and previous studies cholinergic agonists were found to increase the firing of ICC neurons, but the cholinergic agonists were less effective in exciting ICC neurons than EAA agonists. Cholinergic antagonists in a previous study were considerably less effective in inhibiting the discharge of ICC neurons than were the EAA antagonists in the present study. These results, in conjunction with previous neurochemical and anatomical localization studies, support a possible role of an EAA as a candidate for afferent excitatory transmitter in neurons of the inferior colliculus. PMID- 2570055 TI - The quinoxalinediones DNOX, CNOX and two related congeners suppress hair cell-to auditory nerve transmission. AB - We tested 6,7-dinitroquinoxaline-2,3-dione (DNQX); 6-cyano-7-nitroquinoxaline-2,3 dione (CNQX); 6,7-dichloro-3-hydroxy-2-quinoxalinecarboxylic acid (DHQC); and 3 hydroxy-2-quinoxalinecarboxylic acid (3HQC), new kainate and quisqualate receptor antagonists, upon cochlear potentials in guinea pig. Perilymph spaces of guinea pig cochleae were perfused with artificial perilymph solutions containing up to 1000 microM concentrations of DHQC and 3HQC, and 500 microM concentrations of DNQX and CNQX, at a rate of 2.5 microliters/min for 10 min. Cochlear potentials evoked by 10 kHz tone bursts of varying intensity were recorded from the basal turn scala vestibuli. Cochlear perfusion of the four drugs resulted in a dose related suppression of the compound action potential of the auditory nerve (CAP; N1-P1), a prolongation of N1 latency at suprathreshold levels, an elevated CAP threshold, and a decreased N1 latency at CAP threshold. None of the drugs had significant effects on cochlear microphonics (CM) or the summating potential (SP). EC50 values (concentrations causing a 50% reduction in CAP amplitude at 68 dB SPL) were 8 microM for DNQX, 30 microM for DHQC, 35 microM for CNQX, and 1 mM for 3HQC. Results support the hypothesis that kainate and quisqualate receptors are involved in neurotransmission between the hair cell and afferent nerve. PMID- 2570056 TI - Fimbriation genes of Salmonella enteritidis. AB - From a cosmid clone, a 5.3-kilobase (kb) HindIII fragment of Salmonella enteritidis DNA containing the fimA gene was subcloned into bacteriophage T7 promoter vectors in both orientations. Predominantly mature fimbrin (14,000 Mr) was produced by clones containing the 5.3-kb insert, whereas the original cosmid clone predominantly accumulated a prefimbrin precursor (16,500 Mr). T7 RNA polymerase-dependent expression of the 5.3-kb insert and of a series of nested deletions revealed several membrane-localized polypeptides (80,000, 40,000, 29,000, 25,000, and 16,500 Mrs) transcribed in the same direction as fimA as well as a single polypeptide (9,000 Mr) transcribed in the opposite direction. Mini-Mu and TnphoA (Tn5 IS50L::phoA) transposon mutagenesis was used to identify a 2- to 3.5-kb region downstream of fimA that affected fimbrin production and processing. A more distant region (greater than 7 kb), revealed by Tn10 and Mu dI mutagenesis, was also required for fimbriation but did not hybridize with the 5.3 kb fragment. Yet another distant region did hybridize to the 5.3-kb fragment, suggesting the existence of other fimbriation-related genes. PMID- 2570057 TI - Conservation of genomic sequences among isolates of Mycobacterium leprae. AB - Restriction fragment length polymorphism analysis has been used to assess relatedness among the genomes of four isolates of Mycobacterium leprae, the causative agent of leprosy. The M. leprae isolates were from human patients from India, a Mangabey monkey from West Africa, and an armadillo from Louisiana. A total of 16 probes were used; these were insert fragments of M. leprae DNA from plasmid recombinant libraries, 5 of which had genes with identifiable functions and 11 of which were randomly chosen recombinant molecules. In spite of the widely diverse origins of the isolates, restriction fragment length polymorphism analysis demonstrated that less than 0.3% of the nucleotides differ among the genomes. PMID- 2570058 TI - Isolation, characterization, and complementation of Rhizobium meliloti 104A14 mutants that lack glutamine synthetase II activity. AB - The glutamine synthetase (GS)-glutamate synthase pathway is the primary route used by members of the family Rhizobiaceae to assimilate ammonia. Two forms of glutamine synthetase, GSI and GSII, are found in Rhizobium and Bradyrhizobium species. These are encoded by the glnA and glnII genes, respectively. Starting with a Rhizobium meliloti glnA mutant as the parent strain, we isolated mutants unable to grow on minimal medium with ammonia as the sole nitrogen source. For two auxotrophs that lacked any detectable GS activity, R. meliloti DNA of the mutated region was cloned and partially characterized. Lack of cross hybridization indicated that the cloned regions were not closely linked to each other or to glnA; they therefore contain two independent genes needed for GSII synthesis or activity. One of the cloned regions was identified as glnII. An R. meliloti glnII mutant and an R. meliloti glnA glnII double mutant were constructed. Both formed effective nodules on alfalfa. This is unlike the B. japonicum-soybean symbiosis, in which at least one of these GS enzymes must be present for nitrogen-fixing nodules to develop. However, the R. meliloti double mutant was not a strict glutamine auxotroph, since it could grow on media that contained glutamate and ammonia, an observation that suggests that a third GS may be active in this species. PMID- 2570059 TI - Regulation of glutamine synthetase II activity in Rhizobium meliloti 104A14. AB - Most rhizobia contain two glutamine synthetase (GS) enzymes: GSI, encoded by glnA, and GSII, encoded by glnII. We have found that WSU414, a Rhizobium meliloti 104A14 glutamine auxotroph derived from a glnA parental strain, is an ntrA mutant. The R. meliloti glnII promoter region contains DNA sequences similar to those found in front of other genes that require ntrA for their transcription. No GSII was found in the glnA ntrA mutant, and when a translational fusion of glnII to the Escherichia coli lacZ gene was introduced into WSU414, no beta galactosidase was expressed. These results indicate that ntrA is required for glnII expression. The ntrA mutation did not prevent the expression of GSI. In free-living culture, the level of GSII and of the glnII-lacZ fusion protein was regulated by altering transcription in response to available nitrogen. No GSII protein was detected in alfalfa, pea, or soybean nodules when anti-GSII-specific antiserum was used. PMID- 2570061 TI - DNA sequence of the Escherichia coli K-12 gamma-glutamyltranspeptidase gene, ggt. AB - The DNA sequence of ggt, the gene that codes for gamma-glutamyltranspeptidase (EC 2.3.2.2) of Escherichia coli K-12, has been determined. The sequence contains a single open reading frame encoding the signal peptide and large and small subunits, in that order. This result suggests that E. coli gamma glutamyltranspeptidase is processed posttranslationally. PMID- 2570060 TI - Evidence for two different types of insecticidal P2 toxins with dual specificity in Bacillus thuringiensis subspecies. AB - Analysis of polypeptides in the crystalline delta-endotoxins from different Bacillus thuringiensis strains revealed two antigenically similar forms of the P2 protein which differed in molecular mass, peptide profile, and amino acid sequence. Purified preparations of the two forms displayed the characteristic dual toxicity of the P2 protein towards members of the orders Lepidoptera and Diptera in vivo but differed markedly in potency for the insects tested. Both species of the P2 protoxin, solubilized and activated by sequential proteolysis with insect gut extract and alpha-chymotrypsin, retained activity in vivo and in vitro, despite the removal of 144 residues from the N terminus. For the low molecular-mass form, the dual insecticidal activity was reproducible in the in vitro assays. PMID- 2570062 TI - Examination of enterotoxigenic Escherichia coli H10407 (colonization factor antigen I+) by scanning electron microscopy with conductive staining. AB - We have used the scanning electron microscope to examine enterotoxigenic Escherichia coli H10407, which expresses colonization factor antigen I pili. The use of low accelerating voltages and conductive staining procedures allowed us to obtain images of colonization factor antigen I pili and other structural details which were obscured by conventional gold-coating techniques. PMID- 2570063 TI - The current status of neuroleptic therapy. AB - Neuroleptic drugs remain a critical component of the treatment of schizophrenia. Although their use in other conditions such as bipolar disorders and organic mental disorders is also widespread, further research is necessary to validate specific indications and to assess benefit-to-risk ratios. In recent years substantial attention has been focused on establishing minimum effective dosage requirements for both acute and long-term treatment of schizophrenia. Considerable progress has been made in this area. In addition, alternative maintenance-treatment strategies such as targeted or intermittent neuroleptic therapy have been studied. The management of patients who fail to respond to an adequate trial (or trials) of neuroleptic drugs continues to be an enormous clinical challenge. Few data are available on which to base subsequent treatment decisions. Recent research with clozapine suggests that this compound may be helpful in some such individuals, but further efforts to establish other potentially effective treatment strategies for treatment-refractory patients should receive a high priority. PMID- 2570064 TI - Frequency and presentation of neuroleptic malignant syndrome in a state psychiatric hospital. AB - Neuroleptic malignant syndrome, a dangerous complication of antipsychotic drugs, has recently been recognized to occur more commonly than previously thought. However, the frequency of this syndrome has been assessed primarily in academic centers, and these reports may not adequately reflect its frequency in other settings. In a 2-year retrospective assessment of 551 neuroleptic-treated patients in a state psychiatric hospital, the authors diagnosed neuroleptic malignant syndrome in 5 cases (0.9%) using previously defined operational criteria. This observation suggests that the syndrome is probably as common in state facilities as in other institutions. PMID- 2570065 TI - Site-specific mutagenesis of lysine-204, tyrosine-224, tyrosine-228, and histidine-307 of porcine kidney D-amino acid oxidase and the implications as to its catalytic function. AB - In order to evaluate the possible contributions of Lys-204, Tyr-224, Tyr-228, and His-307 in porcine kidney D-amino acid oxidase [EC 1.4.3.3] (DAO) to its catalytic function, we constructed four point mutant cDNAs encoding enzymes possessing Glu-204, Phe-224, Phe-228, and Leu-307 by oligonucleotide-directed in vitro mutagenesis. The four mutant cDNAs and the wild type cDNA could be expressed in vitro with similar efficiencies and about 200 ng of each enzyme protein was produced from 5 micrograms of the respective capped RNA. The electrophoretic mobilities of the in vitro synthesized mutant enzymes on SDS polyacrylamide gel were almost identical with that of the wild type DAO, and the molecular weight was calculated to be 38,000. The Glu-204 and Phe-224 mutant DAOs showed comparable enzyme activities to that of the wild type enzyme, and were inhibited strongly by sodium benzoate, a potent competitive inhibitor of DAO. The kinetic parameters of the two mutant DAOs were also comparable to those of the wild type DAO. On the other hand, the Phe-228 and Leu-307 mutant DAOs showed no detectable activity. The results indicate that Tyr-228 and His-307 play important roles as to the constitution of the active site or participate in the reaction directly, while Lys-204 and Tyr-224 are not essential in the enzyme reaction. PMID- 2570066 TI - Sodium as a mediator of non-phorbol tumor promoter action. Down-modulation of the epidermal growth factor receptor by palytoxin. AB - Previous studies have shown that palytoxin, a non-(12-O-tetradecanoylphorbol-13 acetate)-type tumor promoter, is able to down-modulate the epidermal growth factor (EGF) receptor through a sodium-dependent pathway in Swiss 3T3 cells. A role for sodium is supported by the observation that the sodium proton exchanger monensin and the sodium-conducting ionophore gramicidin mimic palytoxin action by causing a decrease in both high and low affinity EGF binding. However, in addition to causing sodium influx, these agents can induce other cellular effects including changes in membrane polarization, intracellular pH, and macromolecular synthesis. To determine whether any of these factors might be responsible for palytoxin action in our system, we examined the role of each of them in palytoxin induced inhibition of EGF binding. Although palytoxin depolarizes the membrane, the observation that potassium-induced depolarization of the membrane does not cause a decrease in EGF binding, in conjunction with the fact that monensin hyperpolarizes the membrane, indicates that depolarization of the membrane is not responsible for palytoxin-induced changes in the EGF receptor. An investigation of intra-cellular pH suggests that the palytoxin effects are not mediated by proton flux. In addition, nigericin-mediated changes in intracellular pH do not cause an inhibition of EGF binding. Finally, studies conducted in the presence of cycloheximide indicate that protein synthesis is not required for palytoxin action and that inhibition of EGF receptor biosynthesis does not account for palytoxin-induced loss of EGF-binding sites. These results suggest that sodium may act as a second messenger in the signal transduction mechanism by which palytoxin modulates the EGF receptor. PMID- 2570067 TI - Accumulation of cAMP and calcium in S49 mouse lymphoma cells following hyposmotic swelling. AB - Swelling of S49 "wild type" mouse lymphoma cells in hyposmolar medium was used to examine the effects of cellular deformation on cAMP metabolism. In S49 wild type mouse lymphoma cells incubated in a defined medium, progressive reductions in medium osmolarity of 5-50% resulted in proportionate expansion of cell volume. Increases in cell volume were accompanied by incremental increases in intracellular cAMP and calcium. These responses in S49 cells occurred rapidly, with increases in calcium concentration and cAMP content occurring within 1-2 min. Swelling of S49 cells in the absence of ions (hyposmolar versus normosmolar sorbitol) resulted in a significant accumulation of cAMP. Inclusion of papaverine or isobutyl methylxanthine amplified cAMP accumulation, and omission of calcium, sodium, or magnesium from the medium attenuated, but did not prevent accumulation of cAMP in S49 cells in response to swelling. Exposure to propranolol or nadolol attenuated the ability of swelling to increase cAMP concentration, while treatment with 2',5'-dideoxyadenosine or phentolamine had no effect on swelling induced cAMP accumulation. It is concluded that cellular deformation of S49 wild type mouse lymphoma cells stimulates rapid accumulation of intracellular calcium and cAMP. PMID- 2570068 TI - Differentiation-induced gene expression in 3T3-L1 preadipocytes. A second differentially expressed gene encoding stearoyl-CoA desaturase. AB - Previously we isolated and characterized a differentially expressed gene from mouse 3T3-L1 preadipocytes that encodes stearoyl-CoA desaturase (SCD1; Ntambi, J. M., Buhrow, S. A., Kaestner, K. H., Christy, R. J., Sibley, E., Kelly, T. J., Jr., and Lane, M. D. (1988) J. Biol. Chem. 263, 17291-17300). Genomic Southern blot analysis indicated the existence of another closely related gene. Here we report the isolation and characterization of this gene and the corresponding cDNA which encode a second stearoyl-CoA desaturase, SCD2, 3T3-L1 adipocytes. SCD2 cDNA is 5 kilobase pairs in length and encodes a protein of 358 amino acids with greater than 87% amino acid sequence identity to SCD1. RNase protection analysis reveals a 10-fold increase in the expression of SCD2 mRNA during 3T3-L1 preadipocyte differentiation. SCD2 mRNA is expressed constitutively at a high level in brain, is not expressed in liver, and its expression in kidney, adipose, and lung tissue is increased greatly by shifting mice from a diet containing unsaturated fatty acids to a diet devoid of fat. The tissue distribution and the dietary alteration of SCD1 mRNA expression differs markedly from that of SCD2 mRNA being absent from brain, constitutive in adipose tissue, and subject to negative control in liver by feeding a diet containing unsaturated fatty acids. The SCD2 gene spans approximately 15 kilobase pairs and consists of six exons and five introns, with intron/exon junctions similar to those of SCD1. As determined by primer extension analysis the start site of transcription maps 300 nucleotides upstream of the initiator methionine codon. Unlike the SCD1 gene, SCD2 lacks a typical "TATA" box in the 5'-flanking region, but has two "CCAAT" boxes at positions -90 and -135 relative to the transcription initiation site. The SCD2 promoter contains a 140-base pair sequence (located between nucleotides -54 and 201) which possesses 77% sequence identity to a region (located between nucleotides -472 and -325) in the SCD1 promoter. There is a GC-rich sequence in the SCD2 promoter (at nucleotide -175) similar to the binding site for the nuclear transcription factor Sp1 as well as an element with homology to the core consensus sequence for the glucocorticoid regulatory element position -500 and a potential CCAAT box/enhance binding protein sequence at position -540. The SCD gene family provides a new model system for the study of differentiation-induced as well as tissue-specific metabolite controlled gene expression. PMID- 2570069 TI - The importance of the link between Glu204 of the catalytic chain and Arg130 of the regulatory chain for the homotropic and heterotropic properties of Escherichia coli aspartate transcarbamoylase. AB - Recent x-ray crystallographic studies of aspartate transcarbamoylase bound with CTP have detected molecular asymmetry in the interface between the catalytic and regulatory subunits (Kim, K. H., Pan, Z., Honzatko, R. B., Ke, H.-M., and Lipscomb, W. N. (1987) J. Mol. Biol. 196, 863-875). In three of the six interfaces, a salt link occurs between Arg130 of the regulatory chain and Glu204 of the catalytic chain; however, these same residues are 15 A apart in the other three interfaces. In order to determine if this is important for the function of the enzyme, two mutant versions of aspartate transcarbamoylase were created by site-specific mutagenesis. Glu204 of the catalytic chain was converted to a glutamine (Glu204c----Gln) and Arg130 of the regulatory chain was converted to a glycine (Arg130r----Gly). The thermal stability of the Arg130r----Gly enzyme is dramatically reduced, whereas the thermal stability of the Glu204c----Gln enzyme is unaltered compared to the wild-type enzyme. The maximal velocity of both mutant enzymes is identical with that of the wild-type enzyme, however both mutant enzymes have altered substrate affinity and regulatory properties. Based on these studies, the link between Glu204 of the catalytic chain and Arg130 of the regulatory chain is important for the heterotropic properties of the enzyme. Furthermore, the interface between the domain of the regulatory chain which binds zinc and the domain of the catalytic chain which binds aspartate may be more important for CTP inhibition than ATP activation. These data also suggest that heterotropic cooperativity is very sensitive to alterations in the catalytic regulatory interface. However, no clear relationship has been observed between the structural asymmetry and the function of the enzyme. PMID- 2570070 TI - Transepithelial transport of drugs by the multidrug transporter in cultured Madin Darby canine kidney cell epithelia. AB - We studied transepithelial transport of 3H-labeled hydrophobic cationic drugs in epithelia formed by wild-type and by drug-resistant Madin-Darby canine kidney (MDCk) cells that had been infected with a retrovirus carrying the multidrug resistance (MDR1) cDNA which encodes the P-glycoprotein. P-glycoprotein is an ATP consuming plasma membrane multidrug transporter responsible for the efflux of cytotoxic chemotherapeutic drugs from resistant cancer cells. Wild-type MDCK cells have small amounts of P-glycoprotein detected by immunoprecipitation. Net transepithelial transport across wild-type MDCK epithelia was demonstrated. Basal to apical flux of 100 nM vinblastine was about six times higher than apical to basal flux. Addition of unlabeled vinblastine reduced basal to apical flux of tracer and increased apical to basal flux of tracer, a pattern expected if there is a saturable pump that extrudes vinblastine at the apical plasma membrane. Daunomycin, vincristine, and actinomycin D were also actively transported and at 20 microM these agents inhibited transport of vinblastine, suggesting that wild type MDCK cells have a common transporter for all these drugs. Vinblastine transport was also inhibited by 20 microM verapamil, which inhibits the multidrug transporter and reverses multidrug-resistance in non-polarized cells. Net transepithelial transport of all these cytotoxic drugs and of verapamil was much higher in epithelia formed by MDCK cells infected with a human MDR1 virus (MDR MDCK) which is expressed on the apical surface of MDR-MDCK monolayers. Because the transport of these cytotoxic drugs and verapamil is increased in MDR-MDCK epithelia compared to wild-type MDCK epithelia, transport in both these cell populations can be attributed to P-glycoprotein. These results are consistent with a role for P-glycoprotein in multidrug secretory transport across the epithelium of the proximal tubule since P-glycoprotein is normally expressed on the apical membrane of proximal tubule cells. PMID- 2570071 TI - A dipeptide insertion in domain I of exotoxin A that impairs receptor binding. AB - Deletions within the structural exotoxin A gene of 27 or 119 amino acids in domain I of the mature polypeptide, or of 88 or 105 amino acids in domains I and II, resulted in the synthesis of exotoxin A (ETA) polypeptides that were not secreted from Pseudomonas aeruginosa hosts but were localized in the cell membrane. Insertions of a hexanucleotide sequence, either pCGAGCT or pCGAATT, at TaqI sites within the gene resulted in variant exotoxin A polypeptides which were secreted normally. pCGAGCT causes insertion of either Glu-Leu or Ser-Ser in the amino acid sequence of the toxin, while pCGAATT causes insertion of either Glu Phe or Asn-Ser dipeptides. Although the cytotoxicity of eight variants was unimpaired, that of four others was reduced, and one variant which had a Glu-Phe insert between residues 60 and 61 (ETA-60EF61) was 500-fold less cytotoxic than wild-type exotoxin A. Purified ETA-60EF61 dissociated much faster from mouse LMTK cells than wild-type ETA, suggesting that the insertion impaired the ability of ETA-60EF61 to interact with exotoxin A receptors. The location of the insert is within a major concavity on the surface of domain I of the exotoxin A molecule, suggesting that this concavity is important for toxin-receptor interaction. PMID- 2570072 TI - Effects of modifying individual amino or carboxyl groups on the affinity of calmodulin for calcineurin. AB - The effects of modifying individual lysyl, aspartyl, or glutamyl residues in calmodulin on its ability to bind to the neural phosphatase calcineurin have been investigated using a competitive binding method termed "label selection." Samples of calmodulin were radiochemically labeled at a low level (0.03-0.6 group/molecule) by acetylation of amino groups or coupling carboxyl groups with ethanolamine to produce preparations containing predominantly single-site modified and unmodified molecules. These preparations were incubated in a 5-10 fold molar excess with bovine calcineurin under conditions appropriate for complex formation. The bound population was isolated, and the level of modification of each reactive residue was compared with the level in the corresponding group in the intial unselected preparation to determine if molecules modified at specific sites had been selected for or against during the competition for complex formation. Significant selection was observed against molecules modified at Lys21, Asp64, Glu67, Lys75, Glu84, Glu114, Asp118, or Lys148, whereas modification of Glu83 increased binding. The modification of other groups, including components of the four Ca2+-binding sites, had no effect on the interaction. Glu67, a Ca2+-liganding residue in Ca2+-binding site II that may regulate the orientation of this site in relation to the central helix, had the strongest influence on complex formation. Most of the residues identified form a nearly linear array in the three-dimensional structure of calmodulin and indicate the location of an extended surface for interaction with calcineurin and other enzymes. PMID- 2570073 TI - Loss of transcriptional repression of three sterol-regulated genes in mutant hamster cells. AB - Two genes that encode enzymes in cholesterol biosynthesis, 3-hydroxy-3 methylglutaryl-coenzyme A (HMG-CoA) reductase and HMG-CoA synthase, and the gene encoding the low density lipoprotein (LDL) receptor are repressed when sterols accumulate in animal cells. Their 5'-flanking regions contain a common element, designated sterol regulatory element-1 (SRE-1). In the HMG-CoA synthase and LDL receptor promoters, the SRE-1 enhances transcription in the absence of sterols and is inactivated in the presence of sterols. In the HMG-CoA reductase promoter, the region containing the SRE-1 represses transcription when sterols are present. In the current studies, we show that the SRE-1 retains enhancer function but loses sterol sensitivity in mutant Chinese hamster ovary cells that are resistant to the repressor, 25-hydroxycholesterol. In the absence of sterols, the mutant cells produced high levels of all three sterol-regulated mRNAs, and there was no repression by 25-hydroxycholesterol. When transfected with plasmids containing each of the regulated promoters fused to a bacterial reporter gene, the mutant cells showed high levels of transcription in the absence of sterols and no significant repression by sterols. When the SRE-1 in the LDL receptor and HMG-CoA synthase promoters was mutated prior to transfection into the mutant cells, transcription was markedly reduced. Thus, the 25-hydroxycholesterol-resistant cells retain a protein that enhances transcription by binding to the SRE-1 in the absence of sterols, but they have lost the function of a protein that abolishes this enhancement in the presence of sterols. Mutation of a 30-base pair segment of the HMG-CoA reductase promoter that contains the SRE-1 did not reduce transcription in the mutant cells, indicating that this promoter is driven by elements other than the SRE-1. Nevertheless, this promoter failed to be repressed by sterols in the mutant cells. These data suggest that a common factor mediates the effects of sterols on the SRE-1 in all three promoters and that this factor has been functionally lost in the 25-hydroxycholesterol-resistant cells. PMID- 2570074 TI - Characterization of the effects of omega-conotoxin GVIA on the responses of voltage-sensitive calcium channels. AB - 1. omega-conotoxin GVIA (omega-CT) caused a potent (IC50 approximately 2nM) but less than maximal (55%) inhibition of [3H]-noradrenaline release from cortical brain slices induced by K+. At 0.1 microM, omega-CT inhibited [3H] gamma aminobutyric acid (GABA) and [3H]-acetylcholine release by approximately 40%. 2. K+-evoked [3H]-noradrenaline release from cortical brain slices was also characterized with respect to the effects of PN 200-110 (dihydropyridine L channel antagonist), BAY K8644 (L-channel VSCC agonist), and Cd++ (an inorganic L and N-channel antagonist). 10 microM Cd++ and 1 microM PN 200-110 inhibited K+ evoked [3H]-noradrenaline release by 52% and 17%, respectively. 10 microM Bay K 8644 enhanced K+-evoked [3H]-noradrenaline release by 22%, and this enhancement was blocked by 1 microM PN 200-110. 3. omega-CT caused a near-maximal inhibition of the electrically evoked twitch responses of the rat vas deferens (IC50 approximately 10 nM) and guinea-pig ileum (IC50 approximately 60 nM), but had no effect on the postjunctional contractile responses of noradrenaline (vas deferens) or carbachol (ileum). At concentrations as high as 1 microM, omega-CT had no effect on the K+-induced contraction of the rat aorta. 4. Neither the equilibrium binding of [3H]-(+)-PN 200-110 nor the allosteric regulation of [3H] (+)-PN 200-110 binding by tiapamil or diltiazem were altered by omega-CT (0.1 microM). 5. These observations support the notion that the N-type voltage sensitive calcium channel plays a major role in coupling neuronal excitation with neurotransmitter release. PMID- 2570075 TI - The effects of beta-adrenoreceptor antagonists on the force responses of the electrically driven rat right ventricle strip to isoprenaline. AB - 1. The force of the electrically driven rat right ventricle strip is increased by isoprenaline but not by procaterol, a beta 2-adrenoceptor agonist. The effects of some beta-adrenoreceptor antagonists on the responses of the strip to isoprenaline have been determined. 2. Metoprolol (a beta 1-adrenoreceptor selective antagonist) at 3 x 10(-8)-3 x 10(-7) moll-1 and ICI 118,551 (a potent beta 2-adrenoreceptor-selective antagonist) at 10(-7)-10(-6) moll-1 had no effect on maximum responses to isoprenaline and caused parallel rightward shifts of the isoprenaline response curves. The pA2 values for metoprolol and ICI 118,551 were 7.86 and 7.40, respectively. Thus the isoprenaline responses of the rat right ventricle strip are due to stimulation of beta 1-adrenoreceptors. 3. Metoprolol at 10(-6) moll-1 acted as a dual antagonist of the responses of the ventricle strip to isoprenaline causing parallel rightward shifts of the concentration response curves which was due to beta 1-adrenoreceptor antagonism and a depression of the maximum response to isoprenaline which was probably due to membrane stabilizing activity. 4. Low concentrations of (+/-)-pindolol (10(-8) moll-1), (+)-pindolol (10(-7) moll-1), (-)-pindolol (10(-8) moll-1) and mepindolol (10(-10) moll-1) had no effect on isoprenaline maximum responses and caused parallel rightward shifts of isoprenaline response curves. Both (+/-)- and (-)-pindolol at 10(-7) moll-1, mepindolol at 10(-9)-10(-7) moll-1 and bopindolol at 10(-9)-10(-7) moll-1 reduced the isoprenaline maximum responses and caused non parallel rightward shifts of the isoprenaline response curves. 5. BAAM (an irreversible beta-adrenoreceptor antagonist) treatment (3 x 10(-6) moll-1) for 30 min followed by 60 min washout reduced the isoprenaline maximum responses and caused a non-parallel rightward shift of the isoprenaline response curve. The kA (dissociation constant) for isoprenaline was 1.10 x 10(-6) moll-1. Calculation of receptor occupancy demonstrated that in order to produce a maximal response of the rat right ventricle strip, isoprenaline had to occupy 73% of the beta 1 adrenoreceptors. 6. The use of the rat right ventricle strip, a tissue with a small beta 1-adrenoreceptor reserve for isoprenaline, allows a definitive characterization of reversible (e.g. metoprolol) and irreversible (e.g. BAAM) beta 1-adrenoreceptor antagonists. It is suggested that (+/-)- and (-)-pindolol, mepindolol and bopindolol may be slowly dissociating beta 1-adrenoreceptor antagonists. PMID- 2570077 TI - A review of postlysis strategy. AB - The postthrombolysis potential for rethrombosis, unstable angina, and reinfarction is considered, as is adjunctive pharmacologic treatment that may reduce the incidence of such complications. PMID- 2570076 TI - The cortical microfilament system of lymphoblasts displays a periodic oscillatory activity in the absence of microtubules: implications for cell polarity. AB - For an understanding of the role of microtubules in the definition of cell polarity, we have studied the cell surface motility of human lymphoblasts (KE37 cell line) using video microscopy, time-lapse photography, and immunofluorescent localization of F-actin and myosin. Polarized cell surface motility occurs in association with a constriction ring which forms on the centrosome side of the cell: the cytoplasm flows from the ring zone towards membrane veils which keep protruding in the same general direction. This association is ensured by microtubules: in their absence the ring is conspicuous and moves periodically back and forth across the cell, while a protrusion of membrane occurs alternately at each end of the cell when the ring is at the other. This oscillatory activity is correlated with a striking redistribution of myosin towards a cortical localization and appears to be due to the alternate flow of cortical myosin associated with the ring and to the periodic assembly of actin coupled with membrane protrusion. The ring cycle involves the progressive recruitment of myosin from a polar accumulation, or cap, its transportation across the cell and its accumulation in a new cap at the other end of the cell, suggesting an assembly-disassembly process. Inhibition of actin assembly induces, on the other hand, a dramatic microtubule-dependent cell elongation with definite polarity, likely to involve the interaction of microtubules with the cell cortex. We conclude that the polarized cell surface motility in KE37 cells is based on the periodic oscillatory activity of the actin system: a myosin-powered equatorial contraction and an actin-based membrane protrusion are concerted at the cell level and occur at opposite ends of the cell in absence of microtubules. This defines a polarity which reverses periodically as the ring moves across the cell. Microtubules impose a stable cell polarity by suppressing the ring movement. A permanent association of the myosin-powered contraction and the membrane protrusion is established which results in the unidirectional activity of the actin system. Microtubules exert their effect by controlling the recruitment of cytoplasmic myosin into the cortex, probably through their direct interaction with the cortical microfilament system. PMID- 2570078 TI - Determination by coupled high-performance liquid chromatography-gas chromatography of the beta-blocker levomoprolol in plasma following ophthalmic administration. AB - Levomoprolol is a beta-blocking agent used in the topical treatment of glaucoma. The necessity for comparing the plasma levels of a drug administered by the ophthalmic route with those obtained following systemic treatment requires increasingly sensitive methods in order to determine the low plasma concentrations produced by the administration of eye drops. On-line high performance liquid chromatography-gas chromatography and concurrent solvent evaporation proved to be advantageous in the determination of levomoprolol in human plasma. Levomoprolol was determined by capillary gas chromatography (GC) with electron-capture detection (ECD) after solid-phase extraction from plasma and derivatization. Quantitation was based on the internal standard method. The detection limit of 0.2 ng/ml is 50 times lower than that obtained with previous GC methods involving on-column injection and ECD. PMID- 2570079 TI - Application of thermospray liquid chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry for the identification of cynomolgus monkey and human metabolites of SK & F 101468, a dopamine D2 receptor agonist. AB - A combination of thermospray liquid chromatography-mass spectrometry (LC-MS) and LC MS MS has allowed the structural elucidation of a number of metabolites of 4 [2-(dipropylamino)ethyl]-1,3-dihydro-2H-indol-2-one (SK & F 101468) in monkey urine. By using LC-MS-MS with the third quadrupole (Q3) set up in multiple ion detection (MID) mode, a number of metabolites were subsequently detected in the human urine and plasma samples despite very low dosing regimes. This was achieved with minimal sample preparation, e.g. for the urine sample centrifugation was the only preparative step, in order to remove particulate matter, prior to analysis. The good signal-to-noise ratio obtained for the human samples, using LC MS MS with Q3 set up for MID, raised the possibility of a LC-MS-MS quantitative assay. As a result, the detection limit of this method for SK&F 101468 when dissolved in methanol was determined to be in the region of 20 pg on column. PMID- 2570080 TI - Campylobacter species identification based on polymorphism of DNA encoding rRNA. AB - Total DNA from five Campylobacter species was digested with a mixture of XhoI and BglII restriction endonucleases and analyzed by Southern hybridization by using a probe complementary to the DNA coding for the 16S rRNA. Each of the Campylobacter species, including C. jejuni, C. coli, C. laridis, C. fetus, and C. upsaliensis, displayed a characteristic pattern. Although some bands may be common to different species, the simplicity of the hybridization pattern enabled us to discriminate among the different species of Campylobacter. PMID- 2570081 TI - Rapid identification of Streptococcus bovis by using combination constitutive enzyme substrate hydrolyses. AB - Several studies have documented the association of blood and rectal-culture positivity for Streptococcus bovis with gastrointestinal neoplasia, especially colonic carcinoma. Conventional methods using bile-esculin hydrolysis, salt tolerance, and sugar fermentations to differentiate S. bovis from other streptococci are laborious, slow, and relatively expensive. Commercially available systems are costly and require at least 24 to 48 h of incubation. A rapid identification procedure for S. bovis and related bacteria was developed. The method uses a reagent containing two hydrolyzable substrates, p-nitrophenyl alpha-D-galactopyranoside and 4-methylumbilliferyl-beta-D-glucoside, in the presence of 2.5% sodium deoxycholate. This combination test, performed with a rapid assay for L-pyrrolidonyl-aminopeptidase, could distinguish S. bovis, Streptococcus equinus, Enterococcus spp., Streptococcus pneumoniae, and the viridans group streptococci in culture within 30 min. Twelve species of the genera Streptococcus and Enterococcus were tested. The rapid method correlated well with conventional techniques. The reagents are readily available, inexpensive, and easy to make and can be stored in the refrigerator for at least 6 months. PMID- 2570082 TI - Molecular epidemiology of Vibrio cholerae in Hong Kong. AB - We studied restriction fragment length polymorphism of the enterotoxin genes of isolates of Vibrio cholerae El Tor, indistinguishable by bacteriophage typing, which were collected in Hong Kong since 1978. Using this approach, we could distinguish indigenous and exogenous strains obtained from different sources and epidemiological settings. PMID- 2570083 TI - Synchrony and desynchrony in high and low arousal subjects undergoing therapeutic exposure. AB - A comparison of synchrony and desynchrony was made among phobic patients exposed to imaginal flooding under conditions of high and low arousal, respectively. Subjective anxiety (SUDs) and skin conductance (SC) were recorded at five minute intervals during three separate sessions. Results showed some degree of synchrony (small positive relationship) between the two measures for the high arousal group and desynchrony (no relationship) between these measures for the low arousal group. However, with the exception of two cases, one from each group, the findings were by no means clear-cut, suggesting that other factors besides level of arousal may be responsible for individual differences in synchrony. PMID- 2570084 TI - The relationship between neuroleptic drug dose and the performance of psychiatric patients in a maximum security token economy program. AB - This study examined the relationship between neuroleptic medication dose and performance in a token economy program on two maximum security psychiatric wards. Patients receiving higher than average doses exhibited poorer than average program performance but there was a small but statistically significant positive relationship between neuroleptic drug dose (measured in CPZ units/kg) and program performance. However, this positive relationship existed only for the first few weeks of patients' hospital stays, and there was a delay (approximately 2 weeks) between the administration of the drug and the maximal positive effect on program performance. Only a very small minority of medication changes were ever followed by improvements in program performance. The results are discussed in terms of what is a rational strategy for the provision of psychiatric medication and other forms of treatment in institutional settings. PMID- 2570085 TI - Benzodiazepines and pregnancy--more to say and more to learn. PMID- 2570087 TI - Neuroleptic drug levels and antipsychotic effects: a difficult correlation; potential advantage of free (or derivative) versus total plasma levels. AB - Attempts to demonstrate relationships between neuroleptic plasma levels and antipsychotic response have met with mixed results. Eight studies have attempted to clarify such discrepancies by contrasting drug level-response relationships in which the fit of plasma neuroleptic concentrations with response could be contrasted to the fit with antipsychotic response found with free (unbound) neuroleptic (or close derivations of free drug, such as cerebrospinal fluid or erythrocyte neuroleptic concentrations). In describing such drug level-response relationships, seven of eight studies show superiority of free, cerebrospinal fluid or erythrocyte concentrations of drug, as compared with plasma neuroleptic levels (p less than 0.05). Variance between neuroleptic blood levels and therapeutic response was reduced 12.0% +/- 17.7% (SD) by the use of free, cerebrospinal fluid or erythrocyte neuroleptic concentrations rather than plasma drug concentrations. Neuroleptic assays that monitor metabolites as well as parent neuroleptic may also reduce variance between drug level and response. The state of present information is such that routine monitoring of neuroleptic drug concentrations to guide dosage adjustment is premature. PMID- 2570086 TI - A randomized clinical trial of haloperidol decanoate and fluphenazine decanoate in the outpatient treatment of schizophrenia. AB - We carried out an 8-month double-blind clinical trial comparing haloperidol decanoate with fluphenazine decanoate in the maintenance treatment of 72 schizophrenic outpatients. A parallel-groups design was used with stratification by sex and injection interval (2, 3, or 4 weeks). The initial injection interval was based on pretrial maintenance treatment with fluphenazine esters. The dosage equivalency of haloperidol decanoate (1.5 cc or 75 mg) to fluphenazine decanoate (1 cc or 25 mg) used was 3:1. This remained approximately the same throughout a 2 month titration period with a flexible dose regimen, and a further 6-month period with a fixed dose regimen. No statistically significant differences in therapeutic effect were found between the drugs. Both drugs had a similar profile for drug-induced parkinsonism, but there was a trend for differences in masking tardive dyskinesia. Haloperidol and prolactin plasma concentrations were well correlated with dosage, with the exception of haloperidol concentrations in patients receiving injections at 2-week intervals. PMID- 2570088 TI - Efficacy of low-dose metoprolol in neuroleptic-induced akathisia. AB - Recent studies have shown that the beta-blockers can be effective treatments for neuroleptic-induced akathisia. However, the relative contributions of beta-1 versus beta-2 blockade to the therapeutic effect of beta-blockers remains unclear. We treated nine patients who had neuroleptic-induced akathisia with low doses (25-100 mg/day) of the beta-blocker metoprolol. At these doses metoprolol causes selective blockade of beta-1 receptors. Seven patients improved after metoprolol; no further substantial changes were seen after subsequent treatment with propranolol. This finding suggests that neuroleptic-induced akathisia can be improved by selective beta-1 blockade. PMID- 2570089 TI - Prediction of neuroleptic-induced dystonia. PMID- 2570090 TI - Possible enhancing effects of diazepam with reserpine in a chronic schizophrenic patient with tardive dyskinesia. PMID- 2570091 TI - Acute hemodynamic effects of norepinephrine inhibition in patients with severe chronic congestive heart failure. AB - The pathophysiologic role of high levels of circulating catecholamines in patients with congestive heart failure remains unclear. To assess the hemodynamic contribution of circulating catecholamines, metyrosine (alpha-methyl-p-tyrosine), an inhibitor of catecholamine synthesis, was administered to nine patients with acutely decompensated chronic congestive heart failure. Baseline left ventricular ejection fraction averaged 23.3 +/- 9.9%, whereas cardiac output averaged 3.69 +/ 1.03 liters/min, with a pulmonary wedge pressure of 27.4 +/- 8.5 mm Hg. After 48 h of metyrosine administration, plasma norepinephrine concentration decreased from 919.4 +/- 810.6 to 335.4 +/- 143.1 pg/ml (p less than 0.05). Plasma epinephrine concentration averaged 176.4 +/- 166.0 pg/ml at baseline, and was unchanged during metyrosine administration. Despite the significant decrease in circulating norepinephrine, no significant hemodynamic changes were observed during metyrosine administration. These results suggest that high levels of circulating norepinephrine may be more a marker of severe congestive heart failure than an important contributor to the underlying pathophysiology at this advanced stage of the disease process. PMID- 2570092 TI - The infusion of somatostatin reduces the arginine-vasopressin response to insulin induced hypoglycemia in man. AB - The effect of an iv infusion of somatostatin (SRIH) (4.1 micrograms/min x 90 min) on the basal secretion of arginine-vasopressin (AVP) and on the AVP response to insulin (0.15 IU/Kg) - induced hypoglycemia was studied in 6 normal men. Basal AVP secretion was not modified by SRIH administration. The blood glucose decrements induced by insulin were similar in the control insulin-tolerance test (ITT) and in the ITT + SRIH test, whereas the AVP response to hypoglycemia was significantly lower in the presence of SRIH. The mean peak AVP increase was three times higher than the basal value in the control ITT, but only two times during SRIH administration. Infusion of higher doses of SRIH (7 micrograms/min x 90 min) produced similar results. These data suggest the involvement of a somatostatinergic mechanism in regulation of AVP response to hypoglycemia. PMID- 2570094 TI - Effects of physiological increases in plasma noradrenaline on the human endocrine pancreas. AB - The effect of plasma noradrenaline concentrations within the physiological range (less than 5-6 nM) on the endocrine pancreas was investigated in 9 nondiabetic volunteers. Noradrenaline significantly inhibited plasma insulin levels but did not change plasma glucagon and somatostatin concentrations. PMID- 2570095 TI - Glutamate, a neurotoxic transmitter. PMID- 2570093 TI - Endocrine therapy of metastatic breast cancer. PMID- 2570097 TI - An outbreak of viral gastroenteritis in a psychiatric hospital. AB - In September and October 1987, an outbreak of gastroenteritis due to small round structured viruses occurred in a large psychiatric hospital. A total of 149 people, 83 patients and 66 members of staff, were affected over a 26-day period. Most cases were associated with three psychogeriatric wards. The outbreak highlighted the special problems of infection control in long-stay psychiatric units and methods of overcoming these difficulties are discussed. PMID- 2570098 TI - Acinetobacter calcoaceticus biovar anitratus septicaemia in a neonatal intensive care unit: epidemiology and control. AB - Nineteen neonates with septicaemia caused by Acinetobacter calcoaceticus biovar anitratus were treated in a neonatal intensive care unit between October, 1983 and March, 1986. The ages of the patients at the onset of septicaemia ranged from 4 to 22 days (mean 9.7 days). Their birth weights ranged from 1000 g to 3350 g (mean 1790 g) and were less than 2000 g in 14 patients. Antibiotics had been administered to 17 of the 19 neonates before the onset of septicaemia, and all mature infants had received prior antibiotic therapy, intubation or had suffered from a convulsion. Acinetobacter anitratus strains were isolated from pharyngeal swabs and/or faeces from 35 (79.5%) out of 44 infants of less than 2000 g. These strains were also isolated from the hands of staff members, and from equipment such as sinks and baths in the unit. It was likely that nosocomial infection via the hands of the staff occurred. Encouraging frequent hand-washing, strict antibiotic use, and cohorting of colonized infants resulted in a reduction of colonization and no further cases of septicaemia were reported. PMID- 2570096 TI - Peripheral blood and portal tract lymphocyte populations in primary sclerosing cholangitis. AB - The relative distribution of lymphocyte subpopulations in the blood and liver of patients with primary sclerosing cholangitis (PSC) and related diseases has been studied using immunoenzyme techniques. The peripheral blood CD4/CD8 T lymphocyte ratio was significantly higher in active ulcerative colitis (UC) and in PSC with inactive UC than in inactive UC alone. In contrast, no relationship with disease activity was seen in Crohn's disease. The portal tract t lymphocyte count per high power field (mean +/- S.D.) was higher in pre-cirrhotic PSC (173 +/- 105) and primary biliary cirrhosis (PBC: 210 +/- 110) than in histologically normal liver (42 +/- 27). However, the overall portal tract CD4/CD8 ratio was similar in PSC (1.49), PBC (1.89) and normal controls (1.63). The results are consistent with immunological involvement in the pathogenesis of PSC, but argue against the hypothesis that changes in the peripheral blood T cell subsets are due to sequestration at the site of tissue inflammation. PMID- 2570099 TI - Frequency of plasmids in strains of Acinetobacter calcoaceticus. AB - Plasmids were found in 75 out of 93 clinical isolates of Acinetobacter calcoaceticus. The strains harboured up to 20 plasmids each, with two-thirds containing two or more. Most plasmids were smaller than 15 Md. Strains of A. Iwoffi contained more plasmids than A. anitratus. Plasmid analysis may be an easy and valuable tool in the epidemiology of A. calcoaceticus, especially in conjunction with other typing methods. PMID- 2570100 TI - The role of communication in the alteration of patient-care practices in hospital -a prospective study. AB - In infection control, ward practices often need alteration, as in Queen Mary Hospital, Hong Kong, when a policy was introduced to discontinue the recapping of needles among nurses. Nine randomly selected wards were divided into three groups (A, B and C) of 3 wards each. From an initial survey, nurses were divided into those who agreed with the discontinuation (the 'agreeables') and those who did not (the 'non-agreeables'). Methods used to introduce the policy included a simple announcement through the nursing hierarchy in group A (control), a passive method (posters and pamphlets) was added in group B and both passive and active methods (in-service lectures) were used in group C. Five weeks later, behavioural change was assessed by another survey and by an unannounced direct needle-count. Only 21% of the nurses changed their practice by simple announcement. For the 'agreeables', 85% changed by the passive method and no further improvement was observed when the active method was added. However, 83% of the 'non-agreeables' changed their practice when the active method was used while the passive method alone changed only 21%. Six months later, a third survey indicated that when the active method was included the change persisted at over 85%, while a 36% change was noted for the 'agreeables' who were only exposed to the passive method. PMID- 2570101 TI - Hospital use of chlorine disinfectants in a hepatitis B endemic area--a prevalence survey in twenty hospitals. AB - A survey was conducted to assess the uses of chlorine disinfectants in twenty hospitals in Hong Kong. In the 149 areas visited, the charge nurses were interviewed on the use of chlorine disinfectants. A high proportion of uses (44%) were not at the recommended dilution and only 88 (57%) of the 154 samples were within +/- 10% of the manufacturers quoted chlorine content. Samples with inadequate chlorine were found among all six types of chlorine disinfectants, although sodium dichloroisocyanurate tablets conformed to the quoted strength on 88% occasions. Higher usage frequency and better dilution practices were noted for hospitals with a disinfectant policy. PMID- 2570102 TI - Contamination of an endoscope due to Trichosporon beigelli. AB - Trichosporon beigelli was isolated from the gastric aspirates of ten patients who had undergone endoscopy. Suspecting fungal contamination of the endoscopic equipment, samples from the biopsy channel, the water bottle, aspiration catheter and the tip of the endoscope were examined. Only the sample from the biopsy channel showed fungal elements on direct examination and grew Trichosporon beigelli. The original source for the fungus was an immunocompromised patient who had undergone endoscopy earlier. This episode occurred despite adherence to apparently adequate disinfection procedures for endoscopes. Because of the risk of cross-infection, especially in immunocompromised patients, it is imperative to take special steps to prevent fungal contamination of endoscopes. PMID- 2570103 TI - The emergence of methicillin-resistant Staphylococcus aureus infections in an Israeli hospital. AB - Between July 1984 and August 1985, 200 cases of methicillin-resistant Staphylococcus aureus (MRSA) occurred in a large, tertiary care, medical school affiliated hospital. During this period, a limited outbreak, causing serious infections, was identified in the Intensive Care Unit and was contained by appropriate infection control measures. Bacteriophage typing and surveillance cultures failed to identify a common or single source of dissemination of these strains. It appears that MRSA strains have emerged in Israel as endemic pathogens in hospitals, capable of causing serious nosocomial outbreaks. PMID- 2570104 TI - Five days versus one day of penicillin as prophylaxis in elective neurosurgical operations. AB - Of 423 patients undergoing elective cranial and spinal operations, infections due to Staphylococcus aureus occurred in 3 of 217 (1.4%) receiving penicillin for 1 day, in none of 206 receiving penicillin for 5 days. There was no significant difference in rates of infection between the two groups receiving penicillin. It is concluded that penicillin for 1 day is as effective as penicillin for 5 days, in the prevention of wound infections due to S. aureus. PMID- 2570105 TI - Laboratory-acquired brucellosis. AB - Brucellosis is endemic in Saudi Arabia and hospital laboratories are handling increasing numbers of specimens for diagnosis. We report four cases of laboratory acquired brucellosis. PMID- 2570106 TI - Emission of viable bacteria in the exhaust flue gases from a hospital incinerator. AB - The exhaust gases from an oil-fired hospital waste incinerator were examined during normal incinerator operation. The design-specified operating temperature was 800 degrees C in the primary combustion chamber and 1000 degrees C in the secondary chamber. Flue gas temperatures, measured from the sampling point at the base of the exhaust stack, varied over the range 186-305 degrees C, and bacteria were recovered from this position in numbers up to 400 cfu m-3 (mean 56 cfu m-3). No sampling was performed at the top of the stack where flue gases were discharged to the atmosphere. Isolates were predominantly gram positive, i.e. Bacillus spp., coagulase negative staphylococci and Staphylococcus aureus, although low numbers of gram negative species (Pseudomonas fluorescens and other pseudomonads) were also recovered. Our results suggest that incineration may not constitute an absolute method of sterilization for clinical waste. PMID- 2570107 TI - Antibiotic prophylaxis in transurethral resection of the prostate. PMID- 2570108 TI - Use of new broad spectrum antibiotics and the occurrence of fungal endocarditis in a centre for cardiovascular surgery in India. PMID- 2570109 TI - Listeria survives microwave heating. PMID- 2570110 TI - Outbreaks of astrovirus type 1 and rotavirus gastroenteritis in a geriatric in patient population. AB - Two outbreaks of viral gastroenteritis occurred in seven psychogeriatric wards of a 469-bed psychiatric hospital. The outbreaks occurred over an eight-week period; the first affected 30 people and rotavirus was detected in 12 of 14 persons from whom faecal specimens were available. The second affected 32 people (15 of whom were affected in the previous outbreak) and astrovirus was detected in 5 of the 24 people from whom specimens were available. Specific IgM to astrovirus was demonstrated in 3 patients. The rotavirus outbreak was characterised by a longer duration of illness (mean 4 days), and more severe symptoms; two elderly female patients had severe symptoms over a 14-15 day period but recovered. In the astrovirus outbreak the duration of illness was shorter and symptoms were milder although the attack rate was higher than that observed for rotavirus. PMID- 2570111 TI - Romanomermis culicivorax: penetration of larval mosquitoes. AB - In the presence of second larval instars of three mosquito species the preparasites of Romanomermis culicivorax swam near the water surface in an orthokinetic manner. When the preparasites were ca. 1 mm from the host, they stopped and swam klinotactically toward the host. During this phase, the preparasites secreted a small amount of a putative adhesive material from the anterior region and host contact was completed. The adhesive appeared to aid in attachment of the preparasites to the host and initiation of the search-boring phase. The preparasites glided over the host until a suitable penetration site was found. The penetration phase was initiated by probing with the odontostyle. This was followed by partial paralysis, decreased intestinal peristaltic movement, and temporary cardiac arrest in all host mosquitoes which was probably related to injection of esophageal secretions. SEM observations showed that the abdominal walls were the most frequent site for penetration. As the preparasites entered through the penetration hole, microorganisms adhering to the cuticle of the preparasites were retained by the adhesive which accumulated around the penetration site. Thus, microbial contamination of the host was avoided by a mechanical cleansing mechanism. Penetration was usually completed in less than 10 min. PMID- 2570113 TI - Thy-1+ dendritic epidermal cells undergo mitosis in vivo. AB - Recently, morphologic evidence that epidermal Langerhans cells (ELC) undergo a mitotic cycle in normal mouse ear skin has been presented. In the present study, using immunohistochemical staining, we examined the mitotic activity of Thy-1 positive dendritic epidermal cells (Thy-1+DEC) in the normal murine epidermis. A small number of Thy-1+DEC showed round, cleaved, paired, and paired dendritic morphologies, which are identical to those occurring during ELC mitosis. We conclude that normal Thy-1+DEC undergo mitosis within the epidermis to maintain their population in the murine epidermis. PMID- 2570112 TI - Topical cyclosporine A inhibits the phorbol ester induced hyperplastic inflammatory response but not protein kinase C activation in mouse epidermis. AB - Cyclosporine A (CsA) is efficacious in the treatment of psoriasis. Although CsA is known to inhibit T-lymphocyte proliferation in vitro, whether this is its mode of action in psoriasis is uncertain. 12-0-tetradecanoylphorbol-13-acetate (TPA) induces an inflammatory, hyperplastic response in mouse skin, with many of the biochemical and histologic aberrations that occur in psoriatic epidermis. Protein kinase C is the major cellular phorbol ester receptor, and most responses of cells to TPA are mediated by PK-C, which is directly activated by TPA. We therefore have investigated the effects of CsA on pleiotypic responses induced by TPA and whether CsA acts in vivo as a direct inhibitor of PK-C. Simultaneous application of CsA (1.7 mumol) and TPA (10 nmol) to mouse skin significantly reduced inflammatory cell infiltration and increased epidermal thickness induced by TPA treatment alone. CsA had to be applied within 30 min of TPA application in order to have a significant inhibitory effect. Optimal doses of CsA inhibited TPA induced ODC activity, TGase activity, arachidonic acid release, and interleukin-1 beta (IL-1 beta) mRNA to the same degree (approximately 80%), despite measurement at widely different times (30 min-12 h) required to obtain maximal induction by TPA. CsA did not, however, directly inhibit activation of PK-C by TPA. These data demonstrate that CsA blocks the pleiotypic responses of mouse skin to TPA treatment involving biochemical events, inflammatory cell infiltration, and epidermal hyperplasia. The molecular site(s) of action of CsA appears to be distal to the initial activation of PK-C by TPA and clearly inhibits PK-C inducible events. Furthermore, the above data suggest that CsA may directly affect keratinocytes in vivo. PMID- 2570114 TI - [Psychosomatic approach for vulvodynia]. PMID- 2570116 TI - Guidelines for the prevention and control of nosocomial H.I.V. infections. The Hospital Infection Control Group of Thailand. PMID- 2570115 TI - Pathways and regulation of ammoniagenesis by the LLC-PK1 cells in culture. AB - LLC-PK1 kidney epithelial cells grown under the condition of continuous rocking exhibit a variety of differentiated functions of proximal tubular epithelium, including pH-modulated ammoniagenesis. To further determine their value as a model system, we investigated the pathways of ammoniagenesis under both normal conditions and acid-base manipulations. Pulse-chase studies with carbon 14 labeled glutamine demonstrated a marked delay in glutamine conversion to glutamate, indicating that glutamine deamidation is a critical rate-limiting step, and also provided evidence for metabolism of the glutamine carbon skeleton by the tricarboxylic acid cycle. Ammonia and alanine were the predominant nitrogen metabolites of glutamine at all pH conditions, and the stoichiometry suggested that glutamate is metabolized through both glutamate dehydrogenase and glutamate transaminase at pH 7.4. Increased ammonia production in response to a low pH was associated with increased flux through phosphate-dependent glutaminase and the glutamate transamination pathway and was accompanied by a fall in intracellular glutamate and alpha-ketoglutarate concentrations, which was similar to events in the intact kidney. Studies with the inhibitors acivicin and amino oxyacetate suggested that the gamma-glutamyltranspeptidase and glutamine transamination pathways are inconsequential in LLC-PK1 cells. The phosphate dependent glutaminase pathway appears to play a predominant role in the regulation of ammoniagenesis. The similarity in ammonia metabolism with other in vitro and in vivo models suggests that LLC-PK1 cells will be a useful system for investigating renal ammoniagenesis and the intracellular signals that modulate this process. PMID- 2570117 TI - The anterior pituitary content of neuromedin U-like immunoreactivity is altered by thyrotrophin-releasing hormone and thyroid hormone status in the rat. AB - In this study we have examined the effects of TRH, thyroid hormones and dopamine on the rat anterior pituitary content of neuromedin U-like immunoreactivity. Oral administration of TRH (20 mg/100 g per day) to euthyroid animals evoked a fivefold increase in peptide content after 12 days of treatment. This effect was found to be dependent upon circulating levels of thyroid hormone, since administration of TRH to thyroidectomized animals failed to show a similar effect without simultaneous treatment with tri-iodothyronine. The possibility that the TRH-induced increase in anterior lobe neuromedin U content reflected alterations in prolactin secretion or synthetic rate was also examined. Treatment of euthyroid animals with a dopamine agonist and antagonist was, however, without effect. These results demonstrate a unique relationship between TRH and thyroid hormone levels in increasing the anterior lobe content of neuromedin U immunoreactivity. PMID- 2570118 TI - Post-somatostatin rebound secretion of growth hormone is dependent on growth hormone-releasing factor in unrestrained female rats. AB - To examine the characteristics of GH secretion following the termination of the infusion of somatostatin, unrestrained adult female Wistar rats were subjected to repeated infusions of somatostatin separated by 30-min control periods. When somatostatin was infused for 150 min at a dose of 3, 30 or 300 micrograms/kg body wt per h, the magnitude of the rebound GH secretion increased in a dose-dependent manner. The infusion of somatostatin at a dose of 300 micrograms/kg body wt per h for 60, 150 or 240 min progressively augmented the size of the rebound GH secretion. When an antiserum to rat GH-releasing factor (GRF) was injected i.v. 10 min before the end of the infusion, the peak amplitude of the rebound GH secretion (300 micrograms/kg body wt, 150 min) was reduced to less than 20% of that of control rats. The rebound GH secretion (300 micrograms/kg body wt per h, 150 min) was augmented by a bolus injection of human GRF (1 microgram/kg body wt). The combined effect of the end of infusion of somatostatin and a bolus injection of GRF on the amount of GH secreted was additive. The plasma GH response to GRF was completely inhibited when human GRF (3 micrograms/kg body wt per h) and somatostatin (300 micrograms/kg body wt per h) were infused simultaneously for 150 min. The magnitude of the rebound GH secretion following the termination of the co-administration was larger than that following the somatostatin infusion alone, but this rebound was not enhanced by a bolus injection of human GRF.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570119 TI - Cigarette smoke impairment of human lymphocyte function by inhibition of transglutaminase. AB - The in-vitro and in-vivo effects of cigarette smoke were studied in human peripheral blood lymphocytes by applying a method for the capping of beta 2 microglobulin- or phytohaemagglutinin (PHA)-stimulated lymphocyte transformation (measured as (3H)thymidine incorporation) involving the transglutaminase pseudosubstrate monodansylthiacadaverine (MDTC), whose presence resulted in significantly reduced capping and (3H)thymidine incorporation in a concentration dependent manner. The addition of dimethyl sulphoxide-soluble particles from cigarette smoke to lymphocytes in vitro significantly reduced the capping ability and the PHA-induced (3H)thymidine incorporation. Whereas no significant change in MDTC-dependent capping inhibition was seen in lymphocytes from smokers after 10 d abstinence from smoking. there was a marked decrease in (3H)thymidine incorporation in lymphocytes from smokers after smoking three cigarettes following 10 h abstinence. The tentative conclusion is that exposure to cigarette smoke, or smoke extract, impairs MDTC-dependent capping inhibition and PHA stimulated lymphocyte transformation by transglutaminase inhibition. PMID- 2570121 TI - Antihypertensive drug-prescribing patterns of internists and family physicians. AB - Surveys that evaluated antihypertensive drug-prescribing patterns were mailed to 150 family physicians and 150 primary care internists. The initial mailing was followed by a telephone follow-up and a second mailing. Forty-seven percent of family physicians and 41.9% of the internists who were still in practice returned the questionnaire. When asked about their choice of antihypertensive drug therapy for specific patients (based upon age, race, sex, and coexisting disease), the responses of the two specialties were similar. The only statistically significant difference was observed in the responses for a 58-year-old obese white woman with diabetes and renal impairment. In this example, the family physicians were more likely than internists to recommend an angiotensin converting enzyme inhibitor or a beta-blocker (P = .036). This study demonstrates that the majority of physicians individualized initial therapy for hypertension to the specific patient rather than strictly following a stepped-care approach with diuretics or beta-blockers as initial therapy. PMID- 2570120 TI - Reduction of inflammation, tissue damage, and mortality in bacterial meningitis in rabbits treated with monoclonal antibodies against adhesion-promoting receptors of leukocytes. AB - We tested if specific inhibition of recruitment of leukocytes across the blood brain barrier from the vascular compartment to the cerebrospinal fluid (CSF) space reduced tissue damage and improved the outcome of infection in a rabbit model of experimental meningitis. The CD11/CD18 complex of receptors on leukocytes promotes adhesion of these cells to endothelia, a process required for egress of cells into the extravascular space. Intravenous injection of the anti CD18 mAb IB4 effectively blocked the development of leukocytosis in the CSF of animals challenged intracisternally with living bacteria, bacterial endotoxin, or bacterial cell wall. This effect was associated with protection from blood brain barrier injury as measured by exclusion of serum proteins from CSF in mAb-treated animals. The densities of bacteria in CSF and the degrees of bacterial killing due to ampicillin were not affected by the antibody. Animals receiving the antibody experienced a delay in the development of bacteremia and a significantly reduced inflammatory response during ampicillin-induced bacterial killing. Therapy with mAb IB4 prevented development of brain edema and death in animals challenged with lethal doses of Streptococcus pneumoniae. These studies indicate that the major mechanism of leukocyte migration across the blood brain barrier involves the CD11/CD18 receptors and that inflammatory leukocytes recruited by this mechanism are a major cause of blood brain barrier injury and cerebral edema during meningitis. PMID- 2570122 TI - DNA polymorphism haplotype analysis of beta-thalassemia genes in Taiwan. AB - Twenty unrelated families of homozygous beta-thalassemia were collected for restriction polymorphism haplotype study of the beta-thalassemia (beta T) genes. DNA was extracted from WBC of the family members. With Southern blotting and DNA hybridization, restriction fragment length polymorphisms (RFLPs) were studied at 7 sites within the beta-globin gene cluster, including 5' epsilon-HincII, G gamma HindIII, A gamma-HindIII, psi beta-HincII,3' psi beta-HincII, beta-AvaII,3' beta BamHI. From RFLPs of the family members, restriction polymorphism haplotypes of the 40 beta T chromosomes and the 40 normal beta-globin gene (beta A)-bearing chromosomes were constructed. In beta T chromosomes. 26 (65%) belonged to haplotype I (+----++), 12 (30%) to haplotype II (+-----+), 1 (2.5%) to haplotype III (-++-+-+) and 1 (2.5%) to haplotype IV (++---++) which has not been reported in Chinese before. In beta A chromosomes, 12 different haplotypes were found. Seven (17.5%) belonged to haplotype I, 13 (32.5%) to haplotype II, and 6 (15%) to haplotype V (+----+-). The implications of RFLP and haplotyping of beta T genes in terms of molecular defects and prenatal diagnosis are discussed. PMID- 2570123 TI - Normalization of abnormal diurnal weight gain among chronically psychotic geriatric patients. Is abnormal diurnal weight gain a risk factor in chronic psychosis? AB - We studied 20 geriatric and 87 nongeriatric chronically psychotic male inpatients, 16 acutely psychotic male control subjects, and 14 male normal subjects. The subjects were weighed at 7 a.m. and 4 p.m. weekly for 3 weeks. We normalized the diurnal weight gain (NDWG) as a percentage by subtracting the 7 a.m. weight from 4 p.m. weight, multiplying the difference by 100, and then dividing the result by the 7 a.m. weight. NDWG was .509% +/- .337%, 2.209% +/- 1.529%, .631% +/- .405%, and .533% +/- .410%, among the geriatric men, nongeriatric men, control subjects, and normal subjects respectively. Differences in diagnoses and drugs did not explain these findings. We hypothesize that abnormal diurnal weight gain may be a risk factor for premature death among chronically psychotic inpatients. PMID- 2570124 TI - Alpha 1-adrenoreceptor-mediated increase in acetylcholine release in brain slices during morphine tolerance. AB - Norepinephrine, clonidine, and phenylephrine increased the electrically evoked release of endogenous acetylcholine in cortical slices taken from morphine tolerant guinea pigs. This effect was alpha 1-adrenoreceptor mediated and was opposite to the alpha 2-adrenoreceptor-mediated inhibition of acetylcholine release, normally elicited by norepinephrine and clonidine. In the presence of prazosin, clonidine recovered its normal inhibitory properties, suggesting that morphine tolerance induced the appearance of an alpha 1-adrenoreceptor-mediated response that overshadowed, but did not cancel, the still present alpha 2 adrenoreceptor inhibitory control. The attempt to prove the presence of alpha adrenoreceptors on the nerve endings by testing the effect of norepinephrine in synaptosomal preparations (preloaded with [3H]choline and depolarized with KCl and veratridine) was unsuccessful. Therefore the problem of the exact location of this excitatory input remains to be solved. These results confirm previous findings reporting the increase in cortical acetylcholine release induced by the alpha-adrenoreceptor agonists in morphine-tolerant, freely moving guinea pigs and demonstrate that opiate tolerance inverts the direction of the noradrenergic modulation even in the isolated intracortical cholinergic structures. PMID- 2570126 TI - Actions of excitatory amino acids on somatostatin release from cortical neurons in primary cultures. AB - L-Glutamate, N-methyl-D-aspartic acid (NMDA), quisqualate, and kainate were found to increase endogenous somatostatin release from primary cultures of rat cortical neurons in a dose-dependent manner. The rank order of potency calculated from the dose-response curves was quisqualate greater than glutamate = NMDA greater than kainate, with EC50 values of 0.4, 20, and 40 microM, respectively. Alanine, glutamine, and glycine did not modify the release of somatostatin. The stimulation of somatostatin release elicited by L-glutamate was Ca2+ dependent, was decreased by Mg2+, and was blocked by DL-amino-5-phosphonovaleric acid (APV) and thienylphencyclidine (TCP), two specific antagonists of NMDA receptors. The NMDA stimulatory effect was strongly inhibited by APV in a competitive manner (IC50 = 50 microM) and by TCP in a noncompetitive manner (IC50 = 90 nM). The release of somatostatin induced by the excitatory amino acid agonists was not blocked by tetrodotoxin (1 microM), a result suggesting that tetrodotoxin sensitive, sodium-dependent action potentials are not involved in the effect. Somatostatin release in response to NMDA was potentiated by glycine, but the inhibitory strychnine-sensitive glycine receptor did not appear to be involved. Our data suggest that glutamate exerts its stimulatory action on somatostatin release essentially through an NMDA receptor subtype. PMID- 2570125 TI - Alpha 2-adrenergic, kappa-opiate, and P1-purinergic autoreceptors have mutually antagonistic effects: a new regulatory mechanism? AB - Rat cortical synaptosomes prepared on four-step discontinuous Percoll density gradients were loaded with the fluorescent Ca2+-indicator fura-2 to allow measurement of the intrasynaptosomal free calcium concentration ([Ca2+]i). When P1-purinergic, alpha 2-adrenergic, or kappa-opiate agonists were incubated with these synaptosomes for 1 min, there was a highly significant, dose-dependent reduction in [Ca2+]i. The effects of these agonists were blocked by inclusion of appropriate specific antagonists. When alpha 2-adrenergic and P1-purinergic agonists were coincubated, a mutual antagonism of their effects was observed, and, in fact, an increase rather than a decrease in [Ca2+]i was apparent. This mutual antagonism was reversed by addition of either a P1-purinergic or a alpha 2 adrenergic antagonist. Parallel studies in which kappa-opiate and P1-purinergic agonists were coincubated also demonstrated a mutual antagonism between the individual effects that was reversed by prior inclusion of either a kappa-opiate or P1-purinergic antagonist. As these mutually antagonistic effects have been observed between alpha 2-adrenergic, kappa-opiate, and P1-purinergic receptor mediated events, we suggest that this may be a general phenomenon and may be a regulatory mechanism at nerve endings. PMID- 2570127 TI - Ca2+-dependent regulation of presynaptic stimulus-secretion coupling. AB - In the present study, we have investigated the role of Ca2+ in the coupling of membrane depolarization to neurotransmitter secretion. We have measured (a) intracellular free Ca2+ concentration ([Ca2+]i) changes, (b) rapid 45Ca2+ uptake, and (c) Ca2+-dependent and -independent release of endogenous glutamate (Glu) and gamma-aminobutyric acid (GABA) as a function of stimulus intensity by elevating the extracellular [K+] to different levels in purified nerve terminals (synaptosomes) from rat hippocampus. During stimulation, Percoll-purified synaptosomes show an increased 45Ca2+ uptake, an elevated [Ca2+]i, and a Ca2+ dependent as well as a Ca2+-independent release of both Glu and GABA. With respect to both amino acids, synaptosomes respond on stimulation essentially in the same way, with maximally a fourfold increase in Ca2+-dependent (exocytotic) release. Ca2+-dependent transmitter release as well as [Ca2+]i elevations show maximal stimulation at moderate depolarizations (30 mM K+). A correlation exists between Ca2+-dependent release of both Glu and GABA and elevation of [Ca2+]i. Ca2+-dependent release is maximally stimulated with an elevation of [Ca2+]i of 60% above steady-state levels, corresponding with an intracellular concentration of approximately 400 nM, whereas elevations to 350 nM are ineffective in stimulating Ca2+-dependent release of both Glu and GABA. In contrast, Ca2+ independent release of both Glu and GABA shows roughly a linear rise with stimulus intensity up to 50 mM K+. 45Ca2+ uptake on stimulation also shows a continuous increase with stimulus intensity, although the relationship appears to be biphasic, with a plateau between 20 and 40 mM K+.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570128 TI - Antibodies to a synthetic peptide corresponding to a Ser-40-containing segment of tyrosine hydroxylase: activation and immunohistochemical localization of tyrosine hydroxylase. AB - A peptide corresponding to position 32-47 in tyrosine hydroxylase was synthesized (TH-16) and polyclonal antibodies against this peptide were raised in rabbits (anti-TH-16). The effects of anti-TH-16 on modulation of tyrosine hydroxylase activity were investigated. Anti-TH-16 enhanced the enzymatic activity in a concentration-dependent manner, and the antigen TH-16 inhibited the stimulatory activity of the antiserum in a concentration-dependent manner. The activated enzyme had a lower Km app for the cofactor 2-amino-4-hydroxy-6-methyl-5,6,7,8 tetrahydropterin and a higher Vmax app than the nonactivated enzyme. Anti-TH-16 was characterized further by its ability to immunoprecipitate the enzyme activity by labeling tyrosine hydroxylase after Western blotting and by immunohistochemical labeling of catecholaminergic neurons. Anti-TH-16 did not block activation of tyrosine hydroxylase by phosphorylation catalyzed by cyclic AMP-dependent protein kinase. Exposure of the enzyme to anti-TH-16 and subsequent phosphorylation of the enzyme resulted in a greater activation of the enzyme than the sum of activation produced by these two treatments separately. However, the activation was less than additive when the enzyme was first phosphorylated and subsequently exposed to anti-TH-16. The present study demonstrates the utility of anti-TH-16 in investigating the molecular aspects of the enzyme activation. PMID- 2570130 TI - Neurotransmitter amino acid release from synaptosomes. PMID- 2570129 TI - Glutamate-induced increase in intracellular Ca2+ in cerebral cortex neurons is transient in immature cells but permanent in mature cells. AB - The free cytosolic Ca2+ concentration ([Ca2+]i) of cultured cerebral cortex neurons was determined using a fluorescent Ca2+ chelator (Fluo-3) after exposure of the neurons to glutamate. Mature neurons (8 days in culture) responded within 45 s to 100 microM glutamate by an increase in [Ca2+]i from 75 to 340 nM, an increase that during the following 6 min of exposure reached 400 nM. This increase in [Ca2+]i could not be reversed by removal of glutamate. In the absence of extracellular CaCl2, only part of the initial, rapid, glutamate-induced increase in [Ca2+]i was observed in these neurons. In contrast to these findings, neurons cultured for only 2 days (immature neurons) exhibited only a small (from 75 to 173 nM) increase in [Ca2+]i after exposure to 100 microM glutamate, and this rapid increase in [Ca2+]i tended to decline on prolonged exposure to glutamate. Moreover, after removal of glutamate, the increase in [Ca2+]i was fully reversible. Pharmacological characterization of the response to glutamate in mature neurons showed that the N-methyl-D-aspartate (NMDA) receptor antagonists phencyclidine and D-2-amino-5-phosphonovalerate phosphonovalerate blocked 75 and 90%, respectively, of the response, whereas the non-NMDA antagonist 6-cyano-7-nitroquinoxaline-2,3-dione had little effect. PMID- 2570131 TI - Methylmercury-induced movement and postural disorders in developing rat: high affinity uptake of choline, glutamate, and gamma-aminobutyric acid in the cerebral cortex and caudate-putamen. AB - Subcutaneous administration of methylmercuric chloride to neonatal rats resulted in movement and postural disorders during the fourth postnatal week. Sodium dependent high-affinity uptake of radiolabeled choline, glutamate, and gamma aminobutyric acid (GABA) was measured in homogenates of cerebral cortex and caudate-putamen. There was a significant decrease in the uptake of [3H]choline in the cerebral cortex, but not in the caudate-putamen, at the onset of neurological impairment (73-75%) and at one subclinical stage of toxicity (58-64%). No significant differences in [3H]glutamate uptake were detected in either region. The uptake of [3H]GABA in the presence of 1 mM beta-alanine, which was employed to inhibit the glial uptake process, was reduced significantly in both the cerebral cortex and caudate-putamen at the onset of neurological impairment (50 62%) and at one subclinical stage (40-51%). This decrease in [3H]GABA uptake is consistent with the results of previous studies using this animal model, which demonstrated a preferential degeneration of GABAergic neurons in the cerebral cortex and caudate-putamen of methylmercury-treated animals. Because the high affinity uptake of choline is the rate-limiting step for acetylcholine synthesis by cholinergic neurons, the decrease in [3H]choline uptake may reflect an abnormal development of cholinergic innervation of the cerebral cortex. PMID- 2570132 TI - Monoaminergic neurotransmitters in Alzheimer's disease. An HPLC study comparing presenile familial and sporadic senile cases. AB - Norepinephrine, epinephrine, dopamine, serotonin and their major metabolites were measured in 20 regions of the left hemisphere in 4 presenile familial cases of Alzheimer-type dementia and 4 sporadic senile cases. Both groups were compared to values in normal brains obtained in our laboratory. Quantitative determination of the monoamines was performed by HPLC with electrochemical detection. The clinical diagnosis of Alzheimer-type dementia was confirmed by histological examination of the right hemisphere and brain stem. The serotonergic system was dramatically affected in the familial cases with very low or undetectable serotonin concentrations in most cortical and subcortical areas studied and an important cell loss in the nucleus raphe dorsalis, origin of the main ascending serotonergic system. In the senile demented patients the serotonergic deficit is less important but still clearly present. The noradrenergic, adrenergic and dopaminergic systems were less affected by the disease process in senile sporadic as well as in the presenile familial type of Alzheimer's disease. PMID- 2570134 TI - Monitoring in the intensive care unit is not always necessary. AB - Most patients receiving spinal narcotics can be monitored adequately by well trained nurses on postoperative or postdelivery wards. Patients at high risk (e.g., those with preexisting lung disease or many elderly patients) do need monitoring in the intensive care unit. Also requiring special monitoring are patients for whom epidural narcotics alone will not cover their pain, such as young patients with multiple trauma. Patients without these restrictions, however, can be monitored successfully outside the intensive care unit, although the dose of epidural narcotic should be kept as low as possible. PMID- 2570133 TI - Intraspinal opioids: implications for monitoring. Monitoring in the intensive care unit is essential. AB - Patients receiving intraspinal opiates should be monitored in the intensive care unit for at least 24 hours to prevent potentially lethal outcomes. These include respiratory depression caused by sequestration of the morphine in the cerebrospinal fluid and migration of epidural catheters in the subarachnoid or intravascular space. At this time, most hospitals are not equipped or staffed adequately to guarantee the safety of these patients outside the intensive care unit. PMID- 2570135 TI - Pharmacologic management of myocardial ischemia. AB - Pharmacologic intervention is critical to the management of myocardial ischemia. Four classes of drugs are now used in therapeutic regimens for ischemia. beta Adrenergic blockers decrease ischemia by blocking beta 1-mediated inotropic and chronotropic effects, thereby decreasing myocardial oxygen consumption. Calcium channel blockers alter fast- and slow-response action potentials by decreasing transmembrane calcium flux, resulting in decreased conduction velocity and heart rate and prolonged effective refractory period. These effects positively influence ischemia and atrial dysrhythmias. Additionally, these agents decrease excitation-contraction coupling in vascular smooth muscle, resulting in coronary and peripheral vasodilation. Organic nitrates and nitrites decrease vascular smooth muscle contraction and preload, and afterload, precipitating a decrease in myocardial oxygen consumption and ischemia. They also improve blood flow to areas of compromised flow. Thrombolytic agents are administered in acute situations to dissolve the occluding thrombus, but they must be used within a few hours of the injury. These drugs activate the conversion of plasminogen to plasmin, which breaks down the clot into fibrin degradation products. The choice of agents is determined by the etiology of the patient's ischemia, his or her response to the drug, and the presence of concomitant disease processes. PMID- 2570136 TI - Evidence for glycine as an inhibitory neurotransmitter of vestibular, reticular, and prepositus hypoglossi neurons that project to the cat abducens nucleus. AB - The localization and distribution of brain-stem afferent neurons to the cat abducens nucleus has been examined by high-affinity uptake and retrograde transport of 3H-glycine. Injections of 3H-glycine selectively labeled (by autoradiography) only neurons located predominantly in the ipsilateral medial vestibular and contralateral prepositus hypoglossi nuclei, and in the contralateral dorsomedial reticular formation, the latter corresponding to the location of inhibitory burst neurons. The specificity of uptake and retrograde transport of 3H-glycine was indicated by the absence of labeling of the dorsomedial medullary reticular neurons ipsilateral and in close proximity to the injection site, where local uptake by diffusion could have occurred. The selectivity of uptake and transport was demonstrated by the absence of retrograde labeling following injections of 3H-GABA or 3H-leucine into the abducens nucleus. The immunohistochemical localization of glycine and GABA revealed a differential distribution of the 2 inhibitory neurotransmitter candidates in the extraocular motor nuclei. Glycine-immunoreactive staining of synaptic endings in the abducens nucleus was dense with a widespread soma-dendritic distribution but was sparse in the trochlear and oculomotor nuclei. By contrast, GABA-immunoreactive staining within the oculomotor and trochlear nuclei was associated with synaptic endings that were particularly prominent on the somata of motoneurons. GABA immunoreactive staining in the abducens nucleus, however, was sparse. These differences between glycine- and GABA-immunoreactive staining in the extraocular motor nuclei were correlated with differences in the immunoreactivity of axons in the descending (glycine) and ascending (GABA) limbs of the medial longitudinal fasciculus. Glycine-immunoreactive neurons, furthermore, were observed in the same locations as neurons that were labeled autoradiographically by retrograde transport of 3H-glycine from the abducens nucleus. Electrophysiological recordings from abducens motoneurons and internuclear neurons revealed a marked reduction in the slow positivity of the orthodromic extracellular potential elicited by ipsilateral vestibular nerve stimulation following systemic administration of strychnine, an antagonist of glycine. Intracellular recordings demonstrated that the vestibular-evoked disynaptic inhibitory postsynaptic potentials in abducens neurons were effectively blocked by strychnine but were unaffected by picrotoxin, an antagonist of GABA.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2570138 TI - Therapeutic narcotics in the workplace. PMID- 2570137 TI - Altered branching of serotonin-containing neurons in Drosophila mutants unable to synthesize serotonin and dopamine. AB - The anatomy of peripheral serotonin-containing fibers (5-HT fibers) in the gut of wild-type Drosophila larvae was compared to mutants deficient in the gene that encodes the enzyme dopa decarboxylase (DfDdc mutants). The 5-HT fibers, located in the proventriculus and midgut, were visualized immunocytochemically by using a monoclonal antibody against 5-HT. Since DfDdc larvae are devoid of 5-HT and dopamine in the nervous system, the highly selective uptake capability of 5-HT neurons was used to visualize the 5-HT fibers. We found that the absence of 5-HT and dopamine in the nervous system of DfDdc animals does not prevent 5-HT fibers from reaching their appropriate targets. However, these fibers in the mutant show a 2-fold increase in the extent of branching. This effect is specific to 5-HT fibers, since glutamate-like and FMRFamide-like immunoreactive fibers of the proventriculus and midgut remain unaffected in the mutant. Low but detectable levels of dopamine and 5-HT in the CNS are sufficient to prevent the increase in arborization, as indicated by analyses of a temperature-sensitive Ddc allele (Ddcts2), which has very low dopa decarboxylase activity. The abnormally extensive branching of 5-HT fibers also can be partially rescued by feeding DfDdc larvae with dopamine. In contrast, feeding with a 5-HT-containing diet had no effect on the mutant phenotype. Hypotheses that could explain the mutant phenotype are proposed. PMID- 2570139 TI - Functional activity of intestinal epithelium demonstrated by mRNA in situ hybridization. AB - To elucidate the synthetic activity in relation to the morphology of the epithelial cells of the small and large intestine, in situ hybridization with a poly-deoxyribothymidine (poly d(T)) probe was applied to paraffin sections of formalin-fixed blocks. This effectively displays poly-adenylated RNA and, by implication, messenger RNA (mRNA). By minimizing proteinase K pretreatment, the relative concentrations of cellular mRNA were visualized. This revealed minimal mRNA in crypt columnar cells, and maximal mRNA in proliferating cells and in cells showing terminal differentiation. The latter include surface epithelial cells, endocrine cells, Paneth cells, and maturing, but not mature, goblet cells. The goblet cells showing positive hybridization can be regarded as active cells and show characteristic morphology. Such cells are particularly evident in untreated coeliac disease, remitting ulcerative colitis, and transitional mucosa. The proliferating cells showing increased hybridization include normal mitotically active crypt epithelium, regenerating epithelium in ulcerative colitis, adenomatous epithelium, and adenocarcinomatous epithelium. The similarity of hybridization between metaplastic polyp epithelium and surface colonocytes is consistent with the concept that metaplastic polyps are formed of cells showing premature terminal differentiation. PMID- 2570141 TI - Evidence for dual adrenergic receptor regulation of ovine pineal function. AB - Both alpha 1- and beta-adrenergic receptors are present on ovine pineals. In the rat these two receptors interact so that activation of the alpha 1-receptor potentiates the beta-receptor-mediated changes in cyclic AMP and the correlated changes in pineal N-acetyltransferase (EC 2.3.1.87). Here we investigate possible interactions between alpha 1- and beta-receptors through changes in cyclic AMP and assess the importance of each receptor to the melatonin response in ovine pineal punches/slices in vitro. The adrenergic agonists isoproterenol (ISO), noradrenaline (NA), and phenylephrine (PHE) stimulated dose-dependent changes in cyclic AMP with the order of potency of ISO greater than NA greater than PHE, consistent with their relative binding affinities for the beta-receptor. The beta receptor antagonist, propranolol, showed dose-dependent inhibition of the ISO effect, whereas the alpha 1-selective antagonist, prazosin, had no effect. The S shape of the stimulation and inhibition curves for ISO reflects cyclic AMP changes mediated by the beta-receptor only without interaction through the alpha 1-receptor. Each of the adrenergic agonists stimulated indistinguishable dose dependent increases in melatonin release. The ability of PHE to stimulate changes in melatonin release in the absence of concomitant changes in cyclic AMP indicates an important role for the alpha 1-receptor. Prazosin inhibits this response, substantiating this conclusion. However, as propranolol is also inhibitory, it seems that the alpha 1-receptor response is absolutely dependent upon a small level of beta-receptor stimulation, thus providing evidence for adrenergic receptor interaction at a step other than cyclic AMP. PMID- 2570140 TI - Thermal activation of photoactivatable urocanase from Pseudomonas putida. AB - The dark inactivation of urocanase from Pseudomonas putida is caused by the formation of a sulfite adduct of the tightly bound coenzyme, nicotinamide adenine dinucleotide. Photodissociation of this adduct by UV radiation restores the enzyme activity. Based on cold exhaustive dialysis the modification reaction appeared to be irreversible. However, we now report that sulfite modification of urocanase is reversible at higher temperatures. An Arrhenius plot of the thermal activation is linear (20-38 degrees C). The activation energy for the enzyme activation is 114 kJ mol-1. The substance that is photodissociated from inactive urocanase reacts with urocanase to reform the modified enzyme indicating that sulfite is not oxidized, or otherwise changed through these processes. Nucleophiles (sulfite, hydroxylamine, hydride, cyanide) are known to inhibit urocanase by forming adducts with nicotinamide adenine dinucleotide. Urocanase inactivated by hydride or cyanide is not reactivated thermally or photochemically. Urocanase inactivated by hydroxylamine and by glycylglycine can be reactivated by a thermal reaction. In conclusion, sulfite-modified urocanase, which is formed in cells, can be reactivated not only by sunlight but also at physiological temperatures. PMID- 2570142 TI - Quantification and distribution of lymphocyte subsets and Langerhans cells in normal human oral mucosa and skin. AB - Normal human oral (check) mucosa was studied to discover whether the oral cavity resembles the Mucosal Immune System (MIS) or the Skin Immune System (SIS). Immunophenotypes of lymphocyte subsets and Langerhans cells (LC) with their exact locations in the epithelium and papillary layer of the normal buccal mucosa were determined and compared with data of normal human skin. In a double staining procedure, the distribution of T-lymphocytes in relation to blood and lymph vessels was determined. Immunophenotyping of LC was done with a CD1a monoclonal antibody. In contrast to the skin, T-lymphocytes in buccal mucosa are not primarily perivascular in location. They are more or less randomly distributed on both sides of the basement membrane. The epithelium of the buccal mucosa contains about 37 times as many T-lymphocytes as the epidermis of normal skin. T-cell numbers in the papillary layer are more or less comparable. The CD4/CD8 ratios of about 1/2 in the epithelium of buccal mucosa and 1/4 in the skin indicates preferential presence of the CD8 subset in both sites, but the helper/inducer T lymphocytes play a much greater role in the epithelium of the buccal mucosa when compared with skin. B-lymphocytes were not found in the epithelium and papillary layer of the buccal mucosa. Thus, immune response associated cells in buccal mucosa do not show the MIS pattern since B cells are absent. It has more in common with SIS but differences are also apparent. In the epithelium of the buccal mucosa the density of LC does not differ significantly from that of the skin, but the papillary layer of the buccal mucosa contains significantly fewer LC than the skin. As in the skin most of the LC of the buccal mucosa are found in the epithelium. PMID- 2570143 TI - Coated vesicles from the protozoan parasite Trypanosoma brucei: purification and characterization. AB - A procedure was developed to purify a coated vesicle fraction from the protozoan parasite Trypanosoma brucei. Electron microscopy revealed a difference between T. brucei coated vesicles and clathrin-coated vesicles from other eukaryotes: trypanosome vesicles were larger (100 to 150 nm in diameter) and contained an inner coat of electron-dense material in addition to the external coat. Evidence suggests that the internal coat is the parasite's variant surface glycoprotein (VSG) coat. The SDS-PAGE analysis shows the major protein of T. brucei coated vesicles has a molecular mass of 61 kD, similar to VSG; this protein was recognized in an immunoblot by anti-VSG serum. Trypanosome coated vesicles also contain a protein which comigrates with the major protein (clathrin) of coated vesicles purified from rat brains. However, this protein is a minor component and it is not serologically cross-reactive with mammalian clathrin. Immunoblot analysis demonstrated that the parasite vesicles contained host IgG, IgM, and serum albumin. PMID- 2570144 TI - Effects of stimulus timing on transmitter release and postsynaptic membrane potential at crayfish neuromuscular junctions. AB - Different synaptic terminals of the single excitor axon to the opener muscle of crayfish (Procambarus clarkii) often release transmitter in a very different manner when stimulated with the same equal-interval, doublet, or triplet patterns. Compared to synapses that show little facilitation (low Fe synapses), highly facilitating (high Fe) synapses show greater percentage increases in several measures of synaptic efficacy when stimulated with any of these patterns. Low Fe synapses usually show the greater absolute changes in these measures of synaptic efficacy. Changes in the span and pattern of doublets and triplets can independently affect both pre- and postsynaptic measures of synaptic efficacy at either low Fe or high Fe synapses. PMID- 2570145 TI - Microvascular thrombolysis to salvage a free flap using human recombinant tissue plasminogen activator. AB - Nonsurgical thrombolysis using tissue plasminogen activator (t-PA) in conjunction with intravenous heparin infusion was successfully used to salvage a free flap with acutely decreased arterial inflow thought to be due to anastomotic thrombosis. Intravenous infusion (12 mg/hr) of t-PA was stopped after 15 min of therapy when spontaneous bleeding was noted at the groin flap donor site. Concomitant treatment with intravenous heparin resulted in rapid improvement of flap perfusion and ultimately complete survival without surgical intervention. This report suggests that in selected cases, target-specific anticoagulant therapy may be beneficial in salvaging microvascular reconstructions complicated by thrombosis. Further experience is warranted to define more clearly the role, the risks, and the extended benefits of this new mode of nonsurgical therapy. PMID- 2570146 TI - Demonstration of alpha 1-adrenoceptors in rat nasal mucosa. AB - 3H-Prazosin was used to demonstrate alpha 1-adrenoceptors in rat nasal mucosa. Specific binding is saturable and occurs to a homogeneous class of binding sites with high affinity (Kd = 0.07 +/- 0.01 nmol/l and with a receptor density of 0.36 +/- 0.02 pmol/g tissue or 14 +/- 1 fmol/mg protein. Kinetic experiments resulted in a Kd-value of 0.03 nmol/l. The binding is stereoselectively inhibited by epinephrine enantiomers. The antagonist prazosin inhibits 3H-Prazosin binding with high affinity, in contrast to yohimbine, classifying the binding sites as alpha 1-adrenoceptors. Inhibition experiments with WB4101 indicated the presence of alpha 1a- (31 +/- 9%) and alpha 1b-adrenoceptor subtypes in the rat nasal mucosa. The order of potencies of agonists determined in competition experiments was (-)epinephrine greater than (+)epinephrine greater than (-)phenylephrine. PMID- 2570147 TI - Decreased levels of circulating CD4+ T lymphocytes during normal human pregnancy. AB - Peripheral blood mononuclear cell (PBMC) populations during human pregnancy have been investigated by many authors, although the different results obtained, principally in relation to T cells, are very discrepant. In this study we aimed to exclude all the possible causes of these discrepancies: small sample size; diurnal rhythm of CD4+ T cells; smoking habits; haemodilution which occurs during pregnancy and inappropriate statistical analysis; in order to determine whether gestation has a definite effect on PBMC populations. We found that the percentage of CD4+ T lymphocytes decreases in the first and second trimesters, returns to the non-pregnant level in the third trimester and remains there in the postpartum period. The percentages of CD3+ T lymphocytes run parallel to those of CD4+ while CD8+ T lymphocytes do not vary. The proportion of CD16+ cells, which include mature NK cells, diminishes in the second trimester and this reduction is maintained in the third trimester and the puerperium. No variation was found in the other PBMC studied (CD20+ lymphocytes, CD14+ monocytes and D/DR+ cells). When parity was considered no difference was seen between primiparous and multiparous women in any of the cell populations tested. PMID- 2570148 TI - Maternal T cells and human pregnancy outcome. AB - Mothers who had just delivered babies of normal birthweight showed significant reductions in the numbers of circulating T and B lymphocytes and of the CD4 T lymphocyte subset. This is consistent with the immunosuppression that occurs in a normal pregnancy. In contrast, the mothers of low birthweight infants did not show these reductions and had circulating lymphocyte numbers not significantly different from normal controls. This suggests that a component of intra-uterine growth retardation may be immunopathological, owing to a failure of the immunosuppressive mechanisms of pregnancy that might normally prevent the fetus from immunological attack. PMID- 2570149 TI - Takayasu arteritis presenting as retinal and vertebrobasilar ischemia. AB - A young woman presented with a 4-month history of retinal and vertebrobasilar ischemia. Angiography demonstrated narrowing of major branches of the aortic arch. Intractable, severe retroorbital pain of the right eye developed after a middle cerebral artery stroke. During 4 weeks of aggressive immunosuppressive therapy including IV high dose bolus corticosteroids and pulse cyclophosphamide, her neurologic deficit improved transiently, but her retroorbital pain persisted. She died of staphylococcal sepsis and pneumonia. An autopsy demonstrated thrombotic or fibrous occlusion, with minimal inflammation, of extracranial arteries. PMID- 2570150 TI - (8a alpha,12a alpha,13a alpha)-5,8,8a,9,10,11,12,12a,13,13a-decahydro- 3-methoxy 12-(methylsulfonyl)-6H-isoquino[2,1-g][1,6]naphthyridi ne, a potent and highly selective alpha 2-adrenoceptor antagonist. PMID- 2570151 TI - Synthesis and evaluation of N,N-di-n-propyltetrahydrobenz[f]indol-7-amine and related congeners as dopaminergic agonists. AB - An evaluation of 6-[2-(di-n-propylamino)ethyl]indole (4), its rigid analogue N,N di-n-propyl-5,6,7,8-tetrahydrobenz[f]indol-7-amine (5), and some related congeners, for ability to suppress serum prolactin in reserpinized rats, revealed modest biological activity in this in vivo model of dopaminergic activity. Although the indole N-H in these compounds can be considered to be oriented "meta" with respect to the ethylamine side chain, compounds with the indole N-H located in the other "meta" position (i.e. 4-[2-(di-n-propylamino)ethyl]indole (2) or its rigid benz[e]indole analogue 3) were much more potent dopamine agonists. The results argue for a particular orientation of the indole N-H vector. In addition, relatively potent dopamine agonists also resulted when the pyrrole portion of the indole ring was replaced by a methanesulfonamido function, supporting the idea that the indole N-H serves as a hydrogen-bond donor. PMID- 2570152 TI - Benzo- and pyrido-1,4-oxazepin-5-ones and -thiones: synthesis and structure activity relationships of a new series of H1 antihistamines. AB - A series of novel benzo- and pyrido-1,4-oxazepinones and -thiones which represents a new structural class of compounds possessing H1 antihistaminic activity was synthesized, and the SARs were evaluated. The antihistaminic activity was determined by blockade of histamine-induced lethality in guinea pigs. The sedative potential was determined by comparison of the EEG profiles of the compounds with those of known sedating and nonsedating antihistamines. Several of the compounds were shown to possess potent H1 antihistaminic activity and to be free of the cortical slowing with synchronized waves and spindling activity found in the EEG of sedative antihistamines. One compound, 2-[2 (dimethylamino)ethyl]-3,4-dihydro-4-methylpyrido[3,2-f]-1,4- oxazepine-5(2H) thione (rocastine) is currently undergoing clinical evaluation as a nonsedating H1 antihistamine. PMID- 2570153 TI - Introduction of a putative dopaminergic prodrug moiety into a 6,7-substitution pattern characteristic of certain 2-aminotetralin dopaminergic agonists. AB - On the basis of the premise that the dopaminergic agonist profile of 2-(di-n propylamino)-5-hydroxy-6-methyltetralin (1a) is due to in vivo oxidation of the 6 methyl moiety and that 1a may represent a novel prodrug strategy, the vicinal methyl-hydroxyl substitution pattern was incorporated into the 6- and 7-positions of 2-(di-n-propylamino)tetralin to give the 6-methyl-7-hydroxy and 6-hydroxy-7 methyl isomers 8 and 9, respectively. A multistep synthetic approach was devised which permitted preparation of target molecules 8 and 9. Pharmacological data revealed that both target compounds exhibit modest dopamine-like effects in the cardioaccelerator nerve assay in the cat, but neither appeared to be metabolically activated as was the case with 1a. The effects of 9 (but not of 8) were antagonized by pretreatment with haloperidol. Thus, the 5-hydroxy-6-methyl substitution pattern in the 2-aminotetralins remains unique as a dopaminergic agonist prodrug structure. PMID- 2570154 TI - Effects of triturated Culiseta melanura (Diptera: Culicidae) on recovery of eastern equine encephalomyelitis virus. AB - Experiments were done to determine the effect, if any, of larval and adult Culiseta melanura (Coquillett) suspensions on the recovery of eastern equine encephalomyelitis (EEE) virus in baby hamster kidney (BHK) and Vero cell cultures. Although triturated pools of this mosquito reduced the titer of EEE virus added to suspensions, suspensions prepared from as many as 100 Cs. melanura larvae or adults did not reduce titers of virus to undetectable levels. Similarly, the titer of EEE virus in orally infected female Cs. melanura remained detectable when a single infected mosquito was triturated in a pool with 99 uninfected mosquitoes. These results show that in BHK or Vero cell culture assay systems, triturated Cs. melanura do not completely prevent the detection of EEE virus. The results therefore indicate that mosquito suspensions would not interfere with the isolation of virus from field-collected mosquitoes. This conclusion suggests that mosquito suspensions were not a factor in previous studies that were unsuccessful at detecting transovarial transmission of EEE virus by Cs. melanura. PMID- 2570156 TI - A genetic linkage study of facioscapulohumeral (Landouzy-Dejerine) disease with 24 polymorphic DNA probes. AB - From analysis of DNA polymorphisms in a panel of 455 subjects from 25 families with facioscapulohumeral (Landouzy-Dejerine) disease, we have found no evidence for close linkage of the disease at 24 different genetic loci, including one from a candidate chromosomal region. Added to previous data, our results provide direction for future collaborative linkage studies. PMID- 2570155 TI - Linkage analysis in the spinal muscular atrophy type of facioscapulohumeral disease. AB - Facioscapulohumeral disease is probably a heterogeneous disorder. We have ascertained and sampled two multigeneration families with the neurogenic form of this disorder, considered to be a type of spinal muscular atrophy (FSHSMA). The two families have 36 affected members. Linkage studies with 10 expressed and seven DNA restriction fragment length polymorphism (RFLP) markers failed to show significant linkage (Zmax greater than or equal to 3.00). However, two areas of probable linkage were defined on chromosomes 1p and 4q with the markers MNS (Zmax = 1.47 at theta max = 0.10) and PGM1 (Zmax = 0.94 at theta max = 0.001) respectively. We are using additional RFLPs from these and other areas of the human genome to screen these families for linkage to FSHSMA. PMID- 2570157 TI - A study of familial hypercholesterolaemia in Iceland using RFLPs. AB - We have studied 17 unrelated families from Iceland who have familial hypercholesterolaemia (FH), using three different restriction fragment length polymorphisms (RFLPs) of the LDL receptor gene. In one family FH was caused by a 2 kb deletion in the LDL receptor gene in the region of exons 9 to 10. The PvuII (intron 15), NcoI (exon 18), and ApaLI (intron 15) RFLPs were used to determine the haplotypes associated with the defective LDL receptor gene in Iceland. Genotypes were determined in 77 subjects from these 17 families, both FH and non FH. A rare new NcoI RFLP was detected in three subjects. Among the patients, at least four different haplotypes were observed indicating that FH in Iceland is caused by at least four different mutations and is a heterogeneous disease, even in the small, geographically isolated population of Iceland. PMID- 2570159 TI - Akathisia in neuroleptic medicated mentally handicapped subjects. AB - Sixty-six neuroleptic medicated mentally handicapped subjects in a hospital were studied to determine the prevalence of drug-induced akathisia. Tardive dyskinesia was also rated on the AIMS scale. Only motor manifestations of akathisia could be assessed as the subjective component of akathisia was difficult to elicit in this population with difficulties in verbal communication. As all the subjects had been on neuroleptics for at least 3 years, only chronic akathisia could be studied. Five subjects had akathisia. Correlational analysis did not reveal any specific associations with any of the demographic, clinical and pharmacological variables studied. A step-wise multiple regression analysis indicated that younger age could be a predictor. Tardive dyskinesia was associated in two of the subjects. The overall conclusion was that 7% of the subjects had chronic akathisia and no specific risk factors could be identified. PMID- 2570158 TI - Identification of the haplotype pattern associated with the mutant PKU allele in the Gypsy population of Wales. AB - Using the full length cDNA probe, the RFLP haplotype patterns at the phenylalanine hydroxylase locus have been studied in the extensive and highly consanguineous Welsh Gypsy population. The pattern associated with the mutant PKU allele is identical to haplotype 4 in the northern European population. Two children with classical PKU are homozygous for this haplotype. We have tracked the mutant allele through four generations to a great grandfather who died 22 years ago. Both affected children almost certainly have inherited a double dose of the same mutant PKU allele from one common ancestor. It should be possible to identify the specific mutation that is associated with haplotype 4 which results in the more serious form of PKU. PMID- 2570160 TI - Tardive dyskinesia with schizophrenic relapse. AB - A case of a borderline subnormal schizophrenic is described in a patient who developed tardive dyskinesia after 21 years of neuroleptic therapy. However, features of tardive dyskinesia manifested only during the psychotic relapses and disappeared on remission of psychotic symptoms. PMID- 2570161 TI - Complete sequence of a cDNA encoding an active rat choline acetyltransferase: a tool to investigate the plasticity of cholinergic phenotype expression. AB - A cDNA clone encoding the complete sequence of an active rat choline acetyltransferase (ChoAcTase; acetyl-CoA:choline O-acetyltransferase, EC 2.3.1.6) has been isolated. Analysis of the deduced amino acid sequence reveals 85% and 31% identity with the porcine and Drosophila melanogaster enzymes, respectively. To further elucidate the molecular basis of neurotransmitter-related phenotypic plasticity, the expression of ChoAcTase mRNA was compared with that of tyrosine hydroxylase [TH; tyrosine 3-monooxygenase, L-tyrosine, tetrahydropteridine:oxygen oxidoreductase (3-hydroxylating), EC 1.14.16.2], in neurons from superior cervical ganglia grown in the following conditions: 1) normal medium, 2) high K+ medium, and 3) normal medium supplemented with 50% muscle-conditioned medium (CM). TH mRNA was expressed in all three media; its level rose in high K+ and decreased strikingly in the presence of CM. ChoAcTase mRNA could be visualized in CM, but fell to undetectable levels in normal and high K+ media. These results suggest that translational or post-translational mechanisms do not play a major role for the modulation of neurotransmitter-associated phenotype. PMID- 2570162 TI - Isolation, characterization, and application of monoclonal antibodies to rat tyrosine hydroxylase. AB - Tyrosine hydroxylase (TH, tyrosine 3-monooxygenase; EC 1.14.16.2) activity in crude extracts of rat pheochromocytoma, rat brain, and bovine adrenal medulla can be immunoprecipitated in an indirect assay by monoclonal antibodies prepared against partially purified rat pheochromocytoma TH. One of these monoclonal antibodies, TH-2D8-2, can be used for immunocytochemical localization of TH in cell bodies, dendrites, and axons in catecholaminergic neurons (e.g., cells in the substantia nigra) of rat brain and in the cell body, neurites, and growth cones of rat pheochromocytoma cells after treatment with nerve growth factor. When linked to Affi-gel 10, this monoclonal antibody can also be used for immunoaffinity purification of rat and bovine TH. These results suggest that TH 2D8-2 is a valuable reagent with which to investigate the localization, physiological regulation, and function of this important enzyme. PMID- 2570163 TI - Sprouting by intact Helisoma neurons: role for glutamate. AB - We examined the possible relationship between sprouting by intact neurons and the elevation of blood acidic amino acids that is reported to occur in Helisoma in response to osmotic stress. In the present study, buccal ganglion neurons 5 were examined following exposure of animals to conditions that induce estivation, a behavioral state exhibited by these freshwater snails in nature. Surprisingly, sprouting activity occurred in some neurons (31%) in normal animals, this being transiently elevated (to 71%) after 4 days of estivation. Blood acidic amino acids (i.e., glutamate and aspartate) were also elevated under this condition, their levels reaching a plateau on day 4. These observations give rise to two hypotheses: 1) the acidic amino acids can induce sprouting, or 2) these agents can release trophic factors, which, in turn, induce sprouting. The first hypothesis was tested by exposing semi-intact preparations in vitro to a range of concentrations of the acidic amino acids; no promotion of sprouting was observed by either glutamate or aspartate. The second hypothesis was supported, however, by the following observations. First, blood from estivated animals was tested for trophic activity. Such blood (but not that from control animals) could promote sprouting by intact neurons in vitro, a response that depended on factor(s) whose filtration behavior on Amicon membranes suggests an approximate molecular weight of 30-100 kD. Second, since medium conditioned by Helisoma ganglia is known to contain a protein(s) that is necessary for sprouting by isolated neurons, we tested the ability of such medium to promote sprouting by intact neurons. Conditioned medium promoted sprouting by intact neurons in vitro in a manner quantitatively and qualitatively similar to that observed in vivo during estivation. Additionally, sprouting by intact neurons in conditioned medium was also attributable to factor(s) that acted as 30-100 kD molecules on Amicon membranes. To test the ability of the acidic amino acids to release or activate these factors, medium was conditioned for short periods in the presence of glutamate or aspartate; glutamate, but not aspartate, was found to enhance conditioned medium. We conclude that glutamate can promote sprouting by intact neurons, but that its action is indirect and is mediated by macromolecular trophic factors. PMID- 2570164 TI - Evidence from neuronal heterokaryons for a trans-acting factor suppressing Thy-1 expression during neuronal development. AB - The expression of Thy-1 on developing sensory neurons cultured in vitro is strictly regulated and follows a temporal course identical to that observed in situ. We have investigated the role of trans-acting factors in this innate regulation by constructing heterokaryons between mature Thy-1.1+ expressing neurons and immature Thy-1.2- neurons. It has been observed by immunofluorescence that within 16 hr of fusion, Thy-1.1 expression is suppressed in such heterokaryons. However, this suppression is reversible, and after 3-4 days in vitro, at the time at which Thy-1.2 is normally expression, there is re expression of the Thy-1.1 molecule producing Thy-1.1+/Thy-1.2+ co-expressing heterokaryons. As nuclear fusion does not occur, it appears that a developmentally regulated diffusible suppressor molecule is responsible for these observations. PMID- 2570165 TI - Selective elevation of urinary enzyme levels after extracorporeal shock wave lithotripsy. AB - Urinary enzyme testing has been used by many investigators to diagnose and monitor various types of renal injury. Three urinary enzymes, N-acetyl-beta glucosaminidase, beta-galactosidase and gamma-glutamyl transferase were monitored in 17 patients before and after a single, unilateral extracorporeal shock wave lithotripsy treatment. Stones were in the renal pelvis or calices except for 1 treated in situ in the proximal ureter. Urine specimens were collected before extracorporeal shock wave lithotripsy and at 1, 3, 5, 7, 10, 14, 21 and 28 days after treatment. N-acetyl-beta-glucosaminidase and beta-galactosidase levels increased significantly after treatment (p less than 0.05). Gamma-glutamyl transferase levels increased after treatment but this was not statistically significant. All enzyme levels were highest on days 1 and 3 after lithotripsy and returned to baseline by day 28. Factors associated with post-treatment enzyme elevation included female sex, a lower pre-treatment creatinine clearance and stone size greater than 1 cm. These findings indicate that there is a transient selective increase in urinary enzyme excretion after extracorporeal shock wave lithotripsy. PMID- 2570166 TI - Surgical management of undescended testis: retrospective study of potential fertility in 274 cases. AB - To determine the fertility potential of men with an undescended testis a retrospective study was performed on 274 patients by evaluating sperm density and sperm motility, as well as histopathological findings of the seminiferous tubules. The patients, who had been treated surgically when they were 2 to 5 or 9 to 12 years old and who were examined when they were 18 to 39 years old, were divided into 4 groups: group 1 (61 patients) underwent bilateral orchiopexy, group 2 (149) underwent unilateral orchiopexy, group 3 (26) underwent unilateral orchiectomy and group 4 (38) received no surgical treatment for a unilateral undescended testis. Significant differences in the sperm density and motility were detected between group 1 (normal range 0 to 7 per cent) and group 2 or 3 (normal range 72 to 79 per cent), and between group 2 or 3 and group 4 (normal range 42 to 58 per cent). Histopathological differences were significant between group 1 or 2 (Johnsen's score count 6.06 to 6.11) and group 4 (4.72) for the affected side, and between group 2 or 3 (9.09 to 9.20) and group 4 (8.60) for the unaffected side. The results suggest that surgical treatment may not significantly ameliorate the fertility potential of patients with bilateral undescended testes. On the other hand, in patients with unilateral undescended testis an operation not later than at early puberty is advisable to maintain the spermatogenic function in the unaffected testis. It is suggested that some unknown factors relating to the highly impaired unilateral undescended testis may in some way inhibit function of the contralateral unaffected testis. PMID- 2570167 TI - Urinary enzymes and calcium oxalate urolithiasis. AB - Male Sprague-Dawley rats were challenged with various hyperoxaluric agents including ammonium oxalate, hydroxy-L-proline, and ethylene glycol. All treatments resulted in increased urinary oxalate. Associated with hyperoxaluria was an increase in urinary levels of renal enzymes, gamma-glutamyl transpeptidase, N-acetyl-beta-glucosaminidase, and alkaline phosphatase. Most of the rats did not demonstrate any significant change in urinary levels of beta galactosidase. There was a highly significant positive correlation between urinary oxalate and N-acetyl-beta-glucosaminidase. PMID- 2570168 TI - Functional and biochemical alterations in the rabbit urinary bladder following ileocystoplasty. AB - Although the use of ileocystoplasty has increased significantly in recent years, very little is known concerning the smooth muscle properties of the implanted bowel segment. In a previous study, preliminary evidence was presented which indicated that the pharmacological response of the cytoplastic ileal segment to autonomic agonists changed toward that of the bladder. The present study extends and expands these preliminary observations on the physiology and pharmacology of augmentation cystoplasty. Augmentation cystoplasty with detubularized ileum was carried out in 16 rabbits. In vivo and in vitro physiological and pharmacological studies were carried out one and three months after surgery. The results can be summarized as follows: 1) in-vivo CMG at one month was similar to that of the preoperative bladder, but at three months there was a 24% increase in capacity, with the presence of multiple phasic contractions beginning at a volume of approximately 65% of capacity. 2) The frequency and magnitude of spontaneous activity in the cystoplastic ileum did not significantly alter from that of the normal ileum. 3) Cystoplastic ileum responded to muscarinic stimulation differently from the normal ileum. The bladder responded with an increase in the tension whereas the ileum responded with an increase in the frequency and amplitude of phasic contractions. The cystoplastic ileum responded with a pronounced sustained contraction with phasic contractions superimposed. The tonic contraction at three months was of a significantly greater magnitude than that at one month. 4) The qualitative and quantitative response to field stimulation of the cystoplastic ileum was altered from that of the ileum towards that of the bladder. 5) The normal bladder contains greater amount of creatine phosphate and lesser amounts of creatine than the normal ileum. Cystoplasty, after three months induced a change in the ileal segment towards the bladder (increased creatine phosphate and decreased creatine). 6) The normal ileum was found to have greater number of muscarinic receptors than the normal bladder whereas the cystoplastic ileum at three months was intermediate. PMID- 2570170 TI - [Multidrug resistance in acute leukemia]. AB - Despite substantial recent advances in leukemia therapy, most leukemia patients eventually relapse and die of recurrent or refractory leukemia. Important issues are how to treat the patients in relapse of leukemia and how to overcome drug resistant leukemic cells. Multidrug resistance of tumor cells has been vigorously studied in terms of P-glycoprotein, which may play important roles in the transport of antitumor drugs through cell membrane. In the present article, the author has reviewed the recent advances in understanding the pathophysiology of drug-resistance in leukemic cells and discussed the clinical approaches to drug resistant leukemia. PMID- 2570169 TI - From the Centers for Disease Control. Update: HIV-2 infection--United States. PMID- 2570171 TI - Restriction fragment length polymorphisms on the q24-q28 region of X chromosome among Japanese population. AB - Restriction fragment length polymorphisms were studied among the Japanese population using 12 polymorphic DNA probes on the q24-q28 region of X chromosome. The frequency distribution for probes p22-33, p482.6a, p43-15, 52A, pPM101, cX33.2 and cpx234, was the same as that for Caucasians, and that for probes 4D-8 and St14-1 (MspI) was slightly different (p less than 0.05). However, it was quite different (p less than 0.01) for probes p114.12, St14-1 (TaqI), 36B-2 and MN12. Probe p114.12 showed no HindIII polymorphism for the Japanese people. On the contrary, probe MN12, which has a low PIC value (0.15) for Caucasians, was found to be useful for Japanese (PIC value = 0.50). These results suggest that 7 DNA probes (p482.6a, p43-15, 52A, St14-1, p114.12 (BclI), 36B-2 and MN12) are useful (PIC greater than 0.42) for linkage analysis of X-linked disease in Japan. PMID- 2570172 TI - First trimester prenatal diagnosis of haemophilia A using factor VIII gene probe. AB - Accurate first-trimester prenatal diagnosis was achieved in a Japanese haemophilia A family by the use of a restriction fragment length polymorphism (RFLP) located within the F.VIII gene. Since the pregnant woman's heterozygosity for BclI polymorphism in F.VIII/intron 18 (F8A) probe was informative, chorionic villus sampling (CVS) was performed at 9 weeks of gestation. Restriction analysis showed that the fetus was heterozygous for the BclI site and had received a normal paternal X chromosome (0.9 kb) and a normal maternal X (1.2 kb). Therefore, we concluded that the fetus was a non-carrier female. Pregnancy went to term and woman gave birth to an apparently healthy female. At one week after birth a coagulation study confirmed that the newborn infant is not a carrier. The first-trimester prenatal diagnosis of haemophilia A is possible by CVS due to a RFLP in the F.VIII gene. PMID- 2570173 TI - Characterization of a novel alpha 1-adrenoceptor antagonist, SGB-1534, in contractile response of isolated canine arterial and venous smooth muscle to exogenous noradrenaline: comparison with prazosin, phentolamine and yohimbine. AB - The pharmacological properties of SGB-1534, 3-[2-[4-[(o-methoxyphenyl)-1 piperazinyl]ethyl]-2,4(1H,3H)- quinazolinedione monohydrochloride, a selective alpha 1-adrenoceptor antagonist, compared with prazosin, phentolamine and yohimbine, were examined in contractile responses of isolated canine mesenteric arteries and veins and femoral arteries and veins to exogenous noradrenaline. The arteries and veins concentration-dependently contracted when exposed to noradrenaline. The sensitivity to noradrenaline, when compared in terms of pD2 values, was significantly higher in the veins than in the arteries. Phentolamine and yohimbine were competitive antagonists against noradrenaline in the arteries and the veins. SGB-1534 and prazosin caused a parallel shift to the right of the concentration-response curves for noradrenaline only in the arteries: the two antagonists were less effective in the veins than in the arteries when low concentrations of noradrenaline were applied. The pharmacological characteristics of SGB-1534 resemble those of prazosin. The pA2 values for SGB-1534 against noradrenaline in the arteries were much higher than those for prazosin, phentolamine and yohimbine. The result indicates that SGB-1534 may predominantly act upon arterial resistance vessels rather than the venous side, resulting in potent hypotension. PMID- 2570174 TI - Use of cultured renal epithelial cells for the study of cisplatin toxicity. AB - The nephrotoxicity of cisplatin was evaluated in an in vitro system with an established cell line of normal rat kidney, NRK-52E. Leakage of enzymes from the cells into the culture medium increased when they were exposed to 1 microM of cisplatin for 72 hr. The level of lipid peroxides increased in the cells after 48 hr of exposure to cisplatin; the increase was more rapid than the enzyme leakage. This culture system can be used to evaluate drug-induced nephrotoxicity and its mechanism. PMID- 2570175 TI - Selectivity of bunitrolol for beta 1- and beta 2-adrenergic receptors and 5HT1B receptors: assessment by biphasic Scatchard plots and biphasic displacement curve analysis with 125I-iodocyanopindolol and 3H-CGP12177. AB - The preference by bunitrolol for beta 1- and beta 2-adrenoceptors of the rat brain, heart and/or lung was assessed by the radioligand binding assay method with 125I-iodocyanopindolol (125I-ICYP) or 3H-CGP12177. Scatchard plots of 125I ICYP binding in the presence of bunitrolol were found to be non-linear. The inhibition constants (Ki) of bunitrolol for high (beta 2-) and low affinity sites (beta 1-) were: 0.42 +/- 0.16 nM for beta 1 and 3.55 +/- 1.61 nM for beta 2 (beta 1 greater than beta 2), respectively. Displacement experiments conducted with the preparations of the rat brain using 125I-ICYP or with the preparations of the rat heart using 3H-CGP12177 yielded Ki values for bunitrolol of 0.53 +/- 0.20 (beta 1) and 2.37 +/- 0.78 (beta 2) nM for 125I-ICYP binding and 2.01 +/- 0.38 (beta 1) and 12.67 +/- 6.54 (beta 2) nM for 3H-CGP12177 binding. In addition, the Ki value for 5HT1B-receptors assessed in displacement experiments conducted with 125I-ICYP in the presence of 30 microM I-metoprolol in the rat brain was 10.54 +/- 5.92 nM. Thus, bunitrolol is a beta 1-selective antagonist. PMID- 2570176 TI - [Regression of arteriosclerotic lesion of the trunk of the left coronary artery]. PMID- 2570177 TI - Effect of black-pigmented Bacteroides gingivalis on cytotoxic activity of linolenic acid against mouse macrophages. AB - The effects of sonicates of black-pigmented Bacteroides gingivalis (Bg-sonicates) on the cytotoxicity of linolenic acid (LA) against mouse macrophages were examined. Treatment of macrophages with LA alone or in combination with Bg sonicates resulted in the release of lactate dehydrogenase (LDH) from the cells. This observation suggests the cytotoxicity of LA. Also, an increase in thiobarbituric acid-reactive materials was observed in experimental systems of the addition of Bg-sonicates to LA. The cytoplasm of macrophages treated with LA was not stained with May-Grunwald-Giemsa solution, while neither the nucleus nor the cytoplasm was stained when the cells were treated with LA and Bg-sonicates. The above cytotoxicity was nearly abolished when LA and Bg-sonicates were preincubated before the addition to macrophages. B. gingivalis is considered to increase the extent of the cytotoxic effects of LA to the cell nucleus by promoting peroxidation of LA. PMID- 2570179 TI - [Congress of the International Council of Nurses in Seoul. Exchanges across the borders]. PMID- 2570180 TI - In vivo and in vitro ultrastructural alterations induced by human immunodeficiency virus in human lymphoid cells. AB - The ultrastructural alterations induced by human immunodeficiency virus (HIV) in human lymphoid cells have been evaluated. Electron microscopic examination of peripheral blood mononuclear cells (PBMC) from 14 male homosexuals with confirmed acquired immunodeficiency syndrome (AIDS) or AIDS-related complex revealed that tubuloreticular inclusions were present in 5-15% of the cell sections from each case. In 5 of 14 cases, cylindrical confronting lamellae were found in 1-2% of the cell sections. No retrovirus-like particles or surface membrane alterations were detected. Neither of these structural alterations was observed in control PBMC obtained from six HIV-seronegative, hepatitis B virus surface antigen (HBsAg)-positive carriers or in 11 healthy subjects. When primary cultures of CD4+-enriched lymphocytes were infected in vitro with HIV, tubuloreticular inclusions could be detected in 3-10% of the cell sections, but no cylindrical confronting lamellae-like structures were found. In contrast, neither of these alterations were seen in uninfected or HIV-infected H9-HT continuous cell lines. These in vivo and in vitro studies indicate that there is an association between the appearance of the tubuloreticular inclusions and cytopathic HIV infection, although no correlation between cytopathic changes and active viral replication was observed at the single cell level. Further studies will be required to establish the mechanism(s) of formation of the tubuloreticular inclusions and to determine their prognostic potential. PMID- 2570178 TI - [Histamine effects on the heart with special reference to cardiac side effects of H2 receptor antagonists]. AB - The existence of cardiac h1- and h2-receptors is evidenced by pharmacologic studies. Despite of the relatively high content of cardiac histamine it is not clarified whether histamine actually plays a physiologic role - apart from pharmacologic effects - in the regulation of myocardial function and coronary blood flow. Under pathophysiologic conditions (during anaphylaxis, surgical procedures, accidents, stress etc.), however, when a local or systemic histamine release occurs both hemodynamic and arrhythmogenic effects are evident. Numerous studies in animal models conclusively demonstrated a role of cardiac histamine as a major mediator of serious arrhythmias. Consequently, a combination of h1- and h2-receptor antagonists (f.e. Dimetinden/Cimetidin) was recommended as a prophylactic treatment against severe anaphylaxis including life-threatening arrhythmias for cardiac patients at risk. There is pharmacologic evidence of both a positive inotropic and chronotropic effect in the human heart via h2-receptor and stimulation of adenylate cyclase. Histamine-induced coronary effects such as vasoconstriction via h1-receptor and coronary dilatation via h2-receptor are not yet sufficiently validated. Studies on the human heart in vitro using coronary strips from explanted hearts and in vivo investigations on the intact coronary system yielded conflicting results. H2-receptor blocking agents cimetidine, ranitidine and famotidine have qualitatively a different pharmocodynamic spectrum of side effects due to differences in chemical structure. Data on cardiac arrhythmias are mostly associated to cimetidine. Symptomatic bradycardia were reported for both ranitidine and cimetidine. A possible negative inotropic effect of famotidine, although presently not validated, requires further studies. Causative and adverse side effects of cimetidine on the cardiovascular system, however, are to be expected extremely seldom due to easily reversible competetive h2-receptor binding. For prophylaxis rapid intravenous injections of h2-blockers, particularly in elder patients with cardiac diseases, should be avoided. Compared to cimetidine, a tendency of explainable difference seems to become apparent for ranitidine and famotidine due to higher receptor affinity. PMID- 2570181 TI - Myocardial slice: a physiological approach to beta-adrenergic ([3H] CGP-12177) receptor binding in hamster and guinea pig heart. AB - A new technique is described for the characterization and quantification of beta adrenergic receptors in biologically viable slices of myocardium from the hamster right ventricle using the hydrophilic radioligand, [3H]CGP-12177 (CGP). Binding was stereospecific, saturable, of high affinity, reversible, displaceable by appropriate drugs, and highly positively correlated with increasing tissue concentrations. Bmax for CGP binding to myocardial slices from 50-day-old male Golden Syrian hamsters was 3.28 +/- 0.15 fmol/mg wet weight, while Kd was 0.21 +/ 0.02 nM. Freezing resulted in a close to 50% loss of receptor number with no apparent change in affinity. The slice preparation may be utilized to detect in vivo changes in myocardial cell surface receptors, as evidenced by the fact that the number of receptors in slices from ischemic guinea pigs was increased (Bmax = 15.5 +/- 1.25 fmol/mg wet wt) compared with sham-operated controls (Bmax = 10.4 +/- 0.38 fmol/mg wet wt). The minimal tissue disruption associated with this procedure, as well as its speed, simplicity, and relatively low cost, suggest that the myocardial slice preparation provides an important methodology for the study of beta-adrenergic receptor binding in the semiintact myocardium. PMID- 2570182 TI - A grease-gap method for studying the excitatory amino acid pharmacology of CA1 hippocampal pyramidal cells. AB - A grease-gap method for studying the pharmacology of CA1 hippocampal pyramidal cells was developed with use of rat hippocampal slices that included only area CA1 and the retrohippocampal area. These slices were transferred to a two compartment superfusion chamber and the pyramidal cell bodies in area CA1 were separated from their axons in the subiculum with a grease barrier. The CA1 pyramidal cells were depolarized relative to their axons by superfusion with N methyl-D-aspartate (NMDA), (RS)-alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA), kainate and L-glutamate. NMDA was unusually potent in the CA1-subiculum slice compared to other preparations. The NMDA receptor antagonists D(-)-2-amino-5-phosphonovalerate (D-AP5), phencyclidine and Mg2+ shifted the NMDA dose-response curve to the right in a parallel manner. Similarly, the quisqualate receptor antagonist pentobarbitone shifted the AMPA dose-response curve to the right. Schild plots for these antagonists had slopes insignificantly different from 1. These results are consistent with the presence of a substantial NMDA receptor reserve on CA1 pyramidal cells. They are also in line with the high density of excitatory amino acid receptors on CA1 hippocampal pyramidal cells and with the known pharmacological properties of these receptors. Grease-gap studies on the CA1-subiculum slice fill the need for a means of obtaining quantitative pharmacological data on CA1 pyramidal cells. PMID- 2570183 TI - Predictive testing for Huntington's disease with linked DNA markers. AB - Availability of new DNA markers, more tightly linked to the Huntington's disease (HD) locus than the original G8 (D4S10) probes, has improved predictive accuracy for both presymptomatic and prenatal exclusion testing. 50 predictive tests were carried out on high-risk individuals. 6 of these were on first-trimester chorionic villus biopsy specimens; in 2 cases the HD gene was not transmitted to the fetus while in 4 cases no exclusion could be made. The remaining 44 tests were on adults with either 25 or 50% risk of manifesting the disease; 19 had a greatly increased risk and 25 a substantially decreased risk of HD. Family structures in Scotland are suitable for testing about 75% of potentially affected individuals, and the new generation of DNA markers makes virtually all families fully informative. PMID- 2570184 TI - A sicca syndrome in HIV infection: association with HLA-DR5 and CD8 lymphocytosis. AB - A sicca syndrome with parotid enlargement, pulmonary insufficiency, and lymphadenopathy was seen in 12 patients infected with human immunodeficiency virus (HIV), only 1 of whom has had an opportunistic infection during 304 patient months of study. There was a striking increase in numbers of circulating CD8 lymphocytes and the prevalence of HLA-DR5 was greatly increased. In patients with this diffuse infiltrative lymphocytosis syndrome (DILS) the CD8 lymphocytosis, which probably depends on histocompatibility antigen status, may influence disease progression in HIV infection. PMID- 2570185 TI - Carnitine deficiency induced by pivampicillin and pivmecillinam therapy. AB - Short-term administration of pivampicillin and pivmecillinam resulted in a reduction of serum carnitine concentration and an increase in excretion of acylcarnitine in urine. These changes persisted for more than ten days after cessation of therapy. In seven girls on long-term treatment with a mixture of pivampicillin and pivmecillinam the mean total serum carnitine concentration fell to 15% (7-27%) of pretreatment values. The acylcarnitine fraction was 11-57% of total carnitine, compared with less than 2% before treatment. Muscle carnitine concentrations in two girls treated with the antibiotics for 22 and 30 months were only 10% of the mean reference value. These concentrations in serum and muscle are in the range encountered in patients with carnitine deficiencies of other aetiologies in which life-threatening metabolic crises may arise. The risk of adverse effects from prodrugs that give rise to pivalic acid should be seriously considered, particularly in patients under metabolic stress. PMID- 2570186 TI - Probenecid and zidovudine metabolism. AB - The effects of probenecid, a known inhibitor of glucuronidation, on the pharmacokinetics of zidovudine were assessed in eight subjects receiving zidovudine as treatment for human immunodeficiency virus infection. Zidovudine plasma concentrations were measured while subjects were receiving zidovudine alone, after 3 days of zidovudine plus 500 mg probenecid every 8 h, and after 3 days of zidovudine plus 500 mg probenecid plus 260 mg quinine sulphate every 8 h. A median increase of 80% in the area under the zidovudine plasma concentration/time curve occurred with the addition of probenecid. Quinine sulphate prevented the probenecid effect but had no effect on zidovudine kinetics when taken without probenecid by four other subjects. All of the effects were secondary to changes in zidovudine metabolism, since neither probenecid nor quinine changed the renal elimination of zidovudine. Probenecid could be used in combination with zidovudine to extend the interval between doses and reduce the daily requirement for zidovudine, thus enhancing convenience and reducing costs. PMID- 2570187 TI - Cold-induced reversible myocardial ischaemia in systemic sclerosis. AB - The effect of cold provocation on myocardial perfusion was studied in 21 patients with systemic sclerosis and 8 healthy controls. The cold provocation was designed not to cause a pain reaction, and no rise in heart rate/blood pressure product occurred during provocation. Myocardial perfusion was assessed by measurement of thallium uptake by imaged single photon emission computed tomography. No patient had clinical evidence of cardiac involvement, but abnormal electrocardiographic (ECG) findings were found in 5. In 12 patients cold-induced reversible perfusion defects were found; 9 of these also had permanent defects. A further 3 patients had permanent perfusion defects but no reversible defects. The permanent and/or reversible perfusion defects were not related to age among the patients and were not seen in any of the healthy controls, whose age distribution was similar. The reversible and permanent defects were not related to other features of systemic sclerosis, nor to the ECG findings. It is concluded that abnormalities in myocardial perfusion are common in systemic sclerosis and may be present without apparent clinical myocardial involvement. A cold-induced vasopastic process in the myocardial circulation might contribute to the development of the patchy myocardial fibrosis seen in patients with systemic sclerosis. PMID- 2570188 TI - Flecainide and CAST. PMID- 2570189 TI - Are ACE inhibitors safe in pregnancy? PMID- 2570190 TI - Clinical trials of zidovudine in HIV infection. PMID- 2570191 TI - Imaging procedures in the perinatal necropsy. PMID- 2570192 TI - Pseudostatus epilepticus. PMID- 2570193 TI - Compartment syndrome of the thigh. PMID- 2570194 TI - Vitamins and the neural tube. PMID- 2570195 TI - Personal risk-factor chart for cutaneous melanoma. AB - Information from a case-control study of all patients with cutaneous malignant melanoma first diagnosed in Scotland in 1987 has been used to derive a personal risk-factor chart that can be used by both the medical profession and the general public. The relative risk of cutaneous melanoma is estimated from the four strongest risk factors identified by conditional logistic regression. These factors are total number of benign pigmented naevi above 2 mm diameter; freckling tendency; number of clinically atypical naevi (over 5 mm diameter and having an irregular edge, irregular pigmentation, or inflammation); and a history of severe sunburn at any time in life. Use of this risk-factor chart should enable preventive advice for and surveillance of those at greatest risk. PMID- 2570196 TI - 40-year follow-up of overweight children. AB - 504 overweight children admitted to hospital between 1921 and 1947 were followed up for 40 years by questionnaires at 10 year intervals. The mean weight for height (W/H) standard deviation score (SDS) reached a maximum in puberty (+3.5). The SDS fell to about +1 in adulthood. 47% patients were still obese (SDS greater than +1) in adulthood; 84.6% of these had SDS more than +2 in childhood. The degree of obesity in the family (parents and grandparents) and the degree of overweight in puberty were the most important factors for weight level in adulthood. Even when their food intake was in accordance with recommended levels, obese children had higher than normal weight as adults. Excessive overweight in puberty (SDS greater than +3) was associated with higher than expected morbidity and mortality in adult life. Weight-reducing measures should be started early in life to improve the unfavourable long-term prognosis for very obese children. PMID- 2570198 TI - Liver transplantation from live donors. PMID- 2570197 TI - Vasculitis and retinoids. PMID- 2570199 TI - HLA-DQ beta non-ASP-57 allele and incidence of diabetes in China and the USA. PMID- 2570200 TI - Prevention of neural tube defect recurrences in Yorkshire: final report. PMID- 2570201 TI - Diagnostic value of the apex beat. PMID- 2570202 TI - Lowered gastrin and gastric acidity after eradication of Campylobacter pylori in duodenal ulcer. PMID- 2570203 TI - Dispensing hearing aids in the community. PMID- 2570204 TI - Epidemiology of childhood cancer in Zaire. PMID- 2570205 TI - Flunitrazepam in terminal care. PMID- 2570206 TI - Disturbance of cerebral function by aluminium in haemodialysis patients. PMID- 2570207 TI - Prevention of transfusion-acquired cytomegalovirus infection in infants. PMID- 2570208 TI - CpG polymorphisms and haemophilia B. PMID- 2570209 TI - Fish oil in osteoarthritis. PMID- 2570210 TI - Is tannin really bactericidal? PMID- 2570211 TI - Intrathecal baclofen for intractable spasticity due to severe brain injury. PMID- 2570213 TI - Screening for hepatitis C virus antibody. PMID- 2570212 TI - Baroreceptor reset with nitroprusside and drug-resistant hypertension. PMID- 2570214 TI - Time-resolved fluoroimmunoassay for Campylobacter pylori antibodies. PMID- 2570217 TI - French language journals. PMID- 2570215 TI - Geriatric medicine. PMID- 2570216 TI - Two interpretations of drug trial data. PMID- 2570218 TI - Sudden infant death syndrome. PMID- 2570219 TI - Stroke prevention and oestrogen replacement therapy. PMID- 2570220 TI - Ecological impact of antibiotics in neonatal units. PMID- 2570221 TI - Ceftriaxone in bacterial meningitis. PMID- 2570223 TI - Auditory localisation at altitude. PMID- 2570222 TI - Difficulty with Mycobacterium malmoense. PMID- 2570224 TI - Regression of cauda equina syndrome in AIDS patient being treated with ganciclovir. PMID- 2570225 TI - AIDS in Cuba. PMID- 2570226 TI - Benefits of HIV antibody testing in symptom-free risk groups. PMID- 2570228 TI - Cryptogenic meningitis and unexplained episodes of coma in a young child. PMID- 2570227 TI - Prevalence of and mortality from HIV type 2 in Guinea Bissau, West Africa. PMID- 2570229 TI - Captopril and the liver. PMID- 2570231 TI - Corticosteroid administration via artificial caecal fistula in ulcerative colitis. PMID- 2570230 TI - Stage and urinary catecholamine metabolite excretion in neuroblastoma. European Neuroblastoma Study Group. PMID- 2570232 TI - "Indeterminate" western blots and HIV. PMID- 2570233 TI - Effect of non-steroidal anti-inflammatory drugs on the course of osteoarthritis. AB - To test the hypothesis that non-steroidal anti-inflammatory drugs (NSAIDs) accelerate the progression of osteoarthritis by reducing synthesis of vasodilator prostaglandins, thereby diminishing joint perfusion, 105 osteoarthritis patients awaiting hip arthroplasty were treated prospectively with a strong or weak prostaglandin synthesis inhibitor, indomethacin or azapropazone, respectively. Pain and radiological joint space were monitored during the period up to arthroplasty and the condition of the excised femoral head was determined. As judged by radiological and histopathological data, the two treatment groups were at a similar pathophysiological end-point when they came to arthroplasty. In the indomethacin group the affected hips lost joint space more rapidly than did the contralateral hips, a difference not seen in the azapropazone group. The patients receiving azapropazone, who had higher concentrations of synovial vasodilator prostaglandins, took longer than the indomethacin group to reach the arthroplasty end-point. Potent inhibitors of prostaglandin synthesis may be inappropriate in the management of osteoarthritis of the hip. PMID- 2570234 TI - Early suckling and postpartum haemorrhage: controlled trial in deliveries by traditional birth attendants. AB - A randomised, controlled trial was carried out to determine whether suckling immediately after birth reduces the frequency of post-partum haemorrhage (PPH), the mean blood loss, and the frequency of retained placenta. The trial subjects were attended by traditional birth attendants (TBAs), and randomisation was by TBA and not by mother. 68 TBAs attended a course on third stage management and data collection; 19 had to be excluded from the trial. 23 TBAs in the early suckling group and 26 in the control group recorded blood loss in 2104 and 2123 deliveries of liveborn singletons, respectively. The frequency of PPH (loss greater than 500 ml) was 7.9% in the suckling group and 8.4% in the control group and the mean blood loss 258 ml and 256 ml, respectively. Neither of these results differed significantly between the groups. Analysis of the results by individual TBA showed no significant difference between the groups. The frequency of PPH in women of higher parity and in those with multiple pregnancies and stillbirths was high, as expected, which seems to validate the results. The frequency of retained placenta was too low to be analysed. PMID- 2570235 TI - Direct carrier testing in 14 families with haemophilia B. AB - Direct carrier testing was done in 54 at-risk female relatives of haemophilic patients by initially analysing 2.46 kb of the factor IX gene in 1 haemophiliac per family by genomic amplification with transcript sequencing. A presumptive mutation was found in all 14 haemophiliacs examined. Analyses were then done either by sequencing the appropriate region in at-risk female relatives or by detection of an altered restriction site. A simulation indicated that the mutation will be associated with an altered restriction site in about half the families. The technique has clinical application. PMID- 2570236 TI - Endemic cretinism: possible role for thyroid autoimmunity. AB - Thyroid atrophy, rather than goitre, is a characteristic feature of myxoedematous cretinism but its cause and nature are unknown. In this study, purified IgG fractions of serum from patients with myxoedematous endemic cretinism inhibited thyrotropin-induced DNA synthesis in guineapig thyroid segments in a sensitive cytochemical bioassay. IgG from patients with euthyroid neurological endemic cretinism or from normal subjects did not inhibit thyroid growth. Furthermore, in myxoedematous subjects, the presence of the thyroid-growth-blocking immunoglobulins showed a positive relation with thyroid atrophy found on ultrasound. These findings provide a pathogenic basis for the variable clinical expression of endemic cretinism. PMID- 2570237 TI - Diagnosis of beta-thalassaemia by DNA amplification in single blastomeres from mouse preimplantation embryos. AB - Mouse preimplantation embryos were accurately diagnosed as normal or mutant at the beta-major haemoglobin locus by amplification of specific DNA sequences in a single cell. A DNA sequence containing the whole of exon 3 and some 3' untranslated sequences within the beta-major haemoglobin gene was amplified in single blastomeres by means of the polymerase chain reaction (PCR). Blastomeres were removed from embryos of four to eight cells from normal BALB/c mice and from mutant (thalassaemic) BALB/c mice homozygous for a deletion of the whole beta major haemoglobin gene. The sensitivity of the amplification procedure was enhanced by the sequential use of two sets of oligonucleotide primers for 30 cycles of amplification each, the second pair being located within the segment amplified by the first pair. The product (204 base-pairs) could be easily visualised in ethidium bromide-stained agarose gels. Stringent precautions to prevent contamination were taken, and with these precautions the PCR amplification procedure could be carried out under normal laboratory conditions. These procedures for diagnosis of genetic disease before implantation should be applicable to preimplantation diagnosis of any monogenic disorder in man for which the affected DNA sequence is known. PMID- 2570238 TI - Halofantrine in the treatment of malaria. PMID- 2570239 TI - Asthma and the bean. PMID- 2570240 TI - Octreotide. PMID- 2570241 TI - Chromogranin A--always a bridesmaid, never a bride? PMID- 2570242 TI - Shigella dysenteriae type 1 infections in US travellers to Mexico, 1988. AB - In 1988, the number of Shigella dysenteriae type 1 (Sd1) infections reported in the USA increased five-fold over the annual mean from the previous decade. 44 (94%) of 47 interviewed patients reported recent travel to Mexico; 33 (75%) of these had been tourists to the Yucatan peninsula. 27 patients who had travelled to Mexico were admitted to hospital, of whom 2 had a haemolytic uraemic syndrome; none died. The antimicrobial resistance pattern and plasmid profile of the Yucatan strain were similar to those of the 1969-72 pandemic strain. Antimicrobial resistances and plasmid profiles were different in sporadic Western hemisphere strains. This is the first outbreak of Sd1 among US tourists and it is the largest known outbreak in the Western hemisphere since the early 1970s. The dominant Sd1 strain is similar to that which caused the catastrophic 1969-72 pandemic. Surveillance and control measures have been instituted in the Yucatan peninsula. PMID- 2570243 TI - Clinical outcome of coronary angioplasty for single-vessel disease. AB - The acute and long-term results of coronary angioplasty in 412 patients with single-vessel coronary artery disease are reported. Angiographic success was observed in 85.5% of all lesions and 84.7% of all arteries in which dilatation was attempted. Success rates improved during the study; since 1985, the angiographic success rate for non-occluded arteries has been 91.0%. Early complications occurred in 6.8%; angioplasty achieved clinical success in 80.3% of all patients at discharge from hospital. The median duration of follow-up was 772 days, with clinical status established for 98.8% of patients. 5 years after clinically successful angioplasty, cumulative cardiac survival was 96.6% and freedom from cardiac death and myocardial infarction was 90.4% (95.8% and 85.4%, respectively, on intention-to-treat analysis). Within 5 years of successful angioplasty, 12.4% of survivors underwent coronary bypass surgery and 16.9% had repeat angioplasty. Freedom from all cardiac events at 5 years was 70.2% after clinically successful angioplasty and 60.2% on intention-to-treat analysis. Angina symptoms were improved in over 80% of patients and abolished in 75%. PMID- 2570244 TI - Pouting sublinguals: enlarged salivary glands in myxoedema. AB - Enlarged salivary glands are common in patients with hypothyroidism but this finding is not widely appreciated. Although chronic painless enlargement of the salivary glands was reported in papers published 50-60 years ago, mainly in German, it is not mentioned in current textbooks on the thyroid. Along with the parotids, the submandibular and especially the sublingual glands may be quite enlarged, and the enlargement can be a useful clue to the diagnosis of hypothyroidism. The gland enlargement regresses at least partly after thyroid replacement therapy. PMID- 2570245 TI - Western economics and Third World health. PMID- 2570246 TI - Posture and central venous pressure measurement in circulatory volume depletion. PMID- 2570247 TI - Instability of reduced glutathione in commercial Belzer cold storage solution. PMID- 2570248 TI - Q fever outbreak in Birmingham, UK. PMID- 2570249 TI - Trimethoprim resistance and single-agent use. PMID- 2570250 TI - Bacteriotherapy for Clostridium difficile diarrhoea. PMID- 2570251 TI - Staphylococcal carriage and HIV infection. PMID- 2570252 TI - Genital ulcers in women. PMID- 2570253 TI - Inhaled pentamidine in Pneumocystis carinii pneumonia. PMID- 2570254 TI - Reduced CD4+ count, infections, and immune thrombocytopenia without HIV infection. PMID- 2570255 TI - Antibodies to hepatitis C virus in haemophilia. PMID- 2570256 TI - Management of asthma in the community. PMID- 2570257 TI - Screening for antibodies to anaesthetics. PMID- 2570258 TI - Squatting in second stage of labour. PMID- 2570259 TI - Campylobacter pylori and acid secretion. PMID- 2570260 TI - Radon exposure and leukaemia. PMID- 2570261 TI - Baclofen for intractable hiccups. PMID- 2570262 TI - Drugs in donated blood. PMID- 2570264 TI - Informing patients. PMID- 2570263 TI - Late abortions. PMID- 2570265 TI - Learning economics. PMID- 2570266 TI - Medical students' fears about breaking bad news. PMID- 2570267 TI - Cold at heart. PMID- 2570268 TI - Galactose consumption and risk of ovarian cancer. PMID- 2570269 TI - Treatment of haemangioendotheliomas with alpha interferon. PMID- 2570271 TI - Gut perforation after infection with herpes simplex virus and Epstein-Barr virus. PMID- 2570270 TI - Hydrogen cyanide and Bhopal. PMID- 2570272 TI - Propylthiouracil-induced systemic lupus erythematosus. PMID- 2570273 TI - Recombinant human erythropoietin treatment in patients on maintenance home haemodialysis. PMID- 2570274 TI - Detection of Campylobacter pylori infection. PMID- 2570276 TI - Continued function of pancreatic islets after transplantation in type I diabetes. PMID- 2570275 TI - Reduced plasma dehydroepiandrosterone concentrations in Alzheimer's disease. PMID- 2570277 TI - Sudden death and the Cambridge diet. PMID- 2570278 TI - Antibodies to lymphoblastoid interferon. PMID- 2570279 TI - Tryparsamide. PMID- 2570280 TI - Grey mood: grey colours. PMID- 2570281 TI - Toxic epidermal necrolysis and co-trimoxazole. PMID- 2570282 TI - Weight in infancy and death from ischaemic heart disease. AB - Environmental influences that impair growth and development in early life may be risk factors for ischaemic heart disease. To test this hypothesis, 5654 men born during 1911-30 were traced. They were born in six districts of Hertfordshire, England, and their weights in infancy were recorded. 92.4% were breast fed. Men with the lowest weights at birth and at one year had the highest death rates from ischaemic heart disease. The standardised mortality ratios fell from 111 in men who weighed 18 pounds (8.2 kg) or less at one year to 42 in those who weighed 27 pounds (12.3 kg) or more. Measures that promote prenatal and postnatal growth may reduce deaths from ischaemic heart disease. Promotion of postnatal growth may be especially important in boys who weigh below 7.5 pounds (3.4 kg) at birth. PMID- 2570283 TI - Granulocyte-macrophage colony-stimulating factor to harvest circulating haemopoietic stem cells for autotransplantation. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF), given to accelerate recovery from cytopenia induced by high-dose (7 g/m2) cyclophosphamide, reproducibly brought about a dramatic increase (up to 1000-fold) in the number of peripheral blood granulocyte-macrophage colony-forming units (CFU-GM). These circulating progenitors were harvested by leucapheresis and reinfused, together with autologous bone marrow cells, in seven patients with cancer after total body irradiation and melphalan. Complete haemopoietic recovery occurred in all seven transplanted patients in a very short time: mean (SD) 9.1 (0.9) days (range 8-11) to achieve more than 0.5 x 10(9)/l neutrophils, 9.9 (1.7) days (range 8-13) to over 1 x 10(9)/l neutrophils, 10.7 (2.6) days (range 9-16) to over 0.5 x 10(11)/l platelets, and 13.6 (4.2) days (range 13-21) to over 1.0 x 10(11)/l platelets. A reduction in the severity of mucositis was also observed. The rapid haematological recovery made possible by this approach promises to increase the therapeutic index of high-dose chemoradiotherapy regimens and to widen their role as treatment for chemoradiosensitive tumours. PMID- 2570284 TI - Complement deficiencies in patients over ten years old with meningococcal disease due to uncommon serogroups. AB - 46 patients in whom meningococcal disease due to serogroups X, Y, Z, W135, or 29E had developed after the age of 10 years were investigated retrospectively for complement deficiency. Complement deficiency was found in half of the patients: properdin deficiency in 9 patients, C3 deficiency syndromes in 5, and homozygous deficiency of a terminal component (C5, C6, C7, or C8) in 9. Meningococcal infections recurred in 5 of the 9 patients with terminal complement component deficiencies but not in the other complement-deficient patients. The findings show that meningococcal disease due to uncommon serogroups is often associated with complement deficiency. PMID- 2570285 TI - Mutation preventing formation of hepatitis B e antigen in patients with chronic hepatitis B infection. AB - Some patients with chronic hepatitis B virus (HBV) infection are HB e antigen (HBeAg) negative, have circulating HBV particles, and often have especially severe chronic hepatitis. To test the hypothesis that the absence of HBeAg production may be due to a change in the nucleotide sequence of the pre-core region of the genome, 18 Greek and 3 non-Greek patients positive for HB surface antigen underwent direct sequencing of HBV-DNA amplified from sera. In 7 out of 8 HBeAg negative patients, two mutations (guanosine to adenosine) were found in the terminal two codons of the pre-core region, giving the sequence TAGGACATG. The remaining patient had the first mutation only. The sequence TGGGGCATG was found in 4 of 5 of the HBeAg positive patients. The first mutation results in a translational stop codon that is predicted to result in failure to produce HBeAg. The rest of the pre-core region in the HBeAg negative patients was otherwise homologous to that of the HBeAg positive patients and to known sequences. PMID- 2570286 TI - Effects of UK 69 578: a novel atriopeptidase inhibitor. AB - UK 69 578 is a competitive inhibitor of endopeptidase 24.11 (the enzyme that degrades atrial natriuretic factor) in vitro. In vivo, UK 69 578 has renal and cardiovascular effects similar to low-dose atrial natriuretic factor infusion, and may be a useful agent in hypertension and heart failure. PMID- 2570287 TI - Carbamazepine update. PMID- 2570288 TI - Long-term survival in biliary atresia. PMID- 2570289 TI - Epidural morphine, hypertension, and aortic surgery. PMID- 2570290 TI - Screening for congenital CMV. PMID- 2570291 TI - NSAIDs and gut damage. PMID- 2570292 TI - Problems in genetic prediction for Huntington's disease. AB - Experience with nearly 300 applicants for predictive testing for Huntington's disease has shown that apart from the expected problems such as those related to third-party interests and the effects of an adverse test result, there were several less foreseen difficulties. These included the finding that some applicants were already clinically affected, requests for testing of minors, unintentional risk alteration for relatives, and the use of research samples for service purposes. More of the problems involved clinical and counselling aspects rather than laboratory procedures. PMID- 2570293 TI - Uptake of presymptomatic predictive testing for Huntington's disease. AB - Predictive testing by means of gene probes was offered to 110 adults at risk of Huntington's disease (HD). A further 91 individuals spontaneously sought predictive testing. Acceptance rates were highest (85.1%) amongst 47 individuals who spontaneously sought testing and were referred from outside the region, and lowest (15.5%) among the 110 invited to consider predictive testing. Many expressed an interest in predictive testing, then withdrew. Fetal exclusion testing was rarely requested, and then only by individuals who lacked the necessary pedigree structure for predictive testing for themselves. PMID- 2570294 TI - Clinical signs of dehydration in children. AB - 102 children with acute gastroenteritis were thought by the admitting junior doctors to be 5% or more dehydrated. As judged by subsequent weight recovery in hospital, the main indicators of mild to moderate dehydration were decreased peripheral perfusion, deep breathing, decreased skin turgor, high urea, low pH, and a large base deficit; a history of increased thirst was just short of statistical significance. Dehydration was not indicated by a history of oliguria, by the presence of restlessness or lethargy, sunken eyes, dry mouth, or a sunken fontanelle or by the absence of tears. Clinical signs of dehydration became apparent at 3-4% rather than 5% dehydration. The degree of dehydration was overestimated by a mean of 3.2%; this caused unnecessary hospital admissions and overtreatment with intravenous fluid. PMID- 2570295 TI - Hypophosphataemia and phosphaturia in paracetamol poisoning. AB - To find out whether the hypophosphataemia in paracetamol poisoning is due to renal loss of phosphate, serum phosphate concentrations were correlated with indices of hepatotoxicity in 273 patients who had taken an overdose of paracetamol, and the renal handling of phosphate was examined in another 40 patients. Hypophosphataemia was a feature of paracetamol poisoning, whether hepatotoxicity was present or not. It correlated with the degree of hepatic damage and was not influenced by glucose infusions. Serum phosphate correlated with renal threshold phosphate concentration, so renal loss rather than intracellular redistribution of phosphate seems to be the reason for the hypophosphataemia in paracetamol overdose, and it correlates well with other indices of severity of poisoning. PMID- 2570296 TI - Drug trials: who takes the risk? PMID- 2570297 TI - Campylobacter pylori, gastrin, acid secretion, and duodenal ulcers. PMID- 2570298 TI - Syphilis in a Renaissance Italian mummy. PMID- 2570300 TI - Serotonin receptors, buspirone, and premenstrual syndrome. PMID- 2570299 TI - Panic attacks, buspirone, and serotonin function. PMID- 2570301 TI - Nervous reactions after first dose of terfenadine in adults. PMID- 2570302 TI - Percutaneous excimer laser coronary angioplasty. PMID- 2570304 TI - Pamidronate and hypercalcaemia of malignancy. PMID- 2570303 TI - Evaluation of test kits for gonadotropins. PMID- 2570305 TI - Rapid and accurate diagnosis of acute hyperparathyroid crisis. PMID- 2570306 TI - Hymenoptera stings and beta-blockers. PMID- 2570307 TI - Interpretation of blood films in diagnosis of malaria. PMID- 2570308 TI - Health care in transition in Kerala. PMID- 2570309 TI - Interpreting scars. PMID- 2570310 TI - Switzerland's drug surveillance. PMID- 2570311 TI - Turks in Bulgaria. PMID- 2570312 TI - World Medical Association. PMID- 2570313 TI - Computer software in general practice. PMID- 2570314 TI - Time limit for anti-snake venom administration. PMID- 2570316 TI - Psychosis and antituberculosis therapy. PMID- 2570315 TI - Insulin autoantibodies--which method? PMID- 2570317 TI - AIDS after Montreal. PMID- 2570319 TI - Plasma lead and cisplatin. PMID- 2570318 TI - Classical complement pathway in HIV infection. PMID- 2570320 TI - Intestinal "lucid interval" and phytobezoar ileus. PMID- 2570321 TI - Nutritional lessons from peace-time Sierra Leone. PMID- 2570322 TI - Soy formula feeding and immunological response in babies of atopic families. PMID- 2570323 TI - Susceptibility of human herpesvirus 6 to acyclovir and ganciclovir. PMID- 2570324 TI - Clindamycin/primaquine for Pneumocystis carinii pneumonia. PMID- 2570325 TI - Dysgonic fermenter type 2 infection. PMID- 2570326 TI - Cervical screening. PMID- 2570327 TI - Cervical cancer in young Americans. PMID- 2570328 TI - Hyper-IgE syndrome and H2-receptor blockade. PMID- 2570329 TI - Smallpox vaccination for investigators using vaccinia recombinants. PMID- 2570330 TI - Cytomegalovirus, serum beta 2 microglobulin, and progression to AIDS in HIV seropositive haemophiliacs. PMID- 2570332 TI - "Pseudo" status epilepticus. PMID- 2570331 TI - Effect of perioperative blood transfusion on recurrence of Crohn's disease. PMID- 2570333 TI - CSF in neonatal unit. PMID- 2570334 TI - Decision aids and the law. PMID- 2570335 TI - DDS-induced photosensitivity with reference to six case reports. AB - Photosensitivity as an adverse reaction to DDS was recognized in 6 patients of our hospital during the summer of 1988. The clinical manifestations and also the management of those patients are given in detail. All doctors and health workers involved with leprosy need to be aware of such a problem and to take correct decisions after weighing the risk of photosensitivity against the potential benefit of DDS. PMID- 2570336 TI - Dopamine uptake by platelets is selective, temperature dependent and not influenced by the dopamine-D1 or dopamine-D2 receptor. AB - The human platelet, which takes up and releases dopamine, has been proposed as a peripheral model for the study of dopaminergic neurons in the central nervous system (CNS). In addition, the platelet has been shown to possess membrane components with pharmacological properties similar to the dopamine-D1 (DA-D1) and D2 (DA-D2) receptor on dopaminergic neurons. We have therefore studied the specificity of the platelet uptake system for dopamine and, as dopamine uptake comprises both internalised and membrane bound dopamine, the contribution of the DA-D1 and DA-D2 receptor to the uptake of dopamine has been assessed. Significant uptake of 3H-dopamine by platelet rich plasma (PRP) occurred after 10 min incubation at 37 degrees C, uptake being maximal after 90 min. In contrast, at 4 degrees C no uptake of 3H-dopamine occurred up to 60 mins incubation but at 20 degrees C was approximately 8% of the 60 min uptake at 37 degrees C. The neurotransmitters serotonin and dopamine inhibited dopamine uptake by platelets in a dose dependent manner. Uptake of dopamine appeared to be via two systems, one of high affinity with low capacity and the other of lower affinity but high capacity. In contrast, noradrenaline, adrenaline, acetylcholine, gamma aminobutyric acid and histamine (10 microM) had no effect on dopamine uptake by platelets. The DA-D1 receptor antagonist SCH 23390 (10 microns) and the DA-D2 receptor antagonists (10 microM) spiperone, domperidone and (+)-butaclamol did not significantly affect dopamine uptake by platelets. In addition, ouabain and desipramine (100 microM) inhibited dopamine uptake by 21% and 24% respectively whilst reserpine and imipramine (100 microM) increased uptake by 14% and 15%. We therefore conclude that platelets take up dopamine via a selective, temperature dependent mechanism. Our data also suggest that dopamine uptake by platelets does not involve the DA-D1 or DA-D2 receptor. PMID- 2570337 TI - Effect of ATP and amiloride on ANF binding and stimulation of cyclic GMP accumulation in rat glomerular membranes. AB - We investigated ANF binding and stimulation of cGMP accumulation in isolated rat glomerular membranes in the presence and absence of amiloride and ATP. Amiloride enhanced high affinity binding of ANF without affecting its stimulation of cGMP. In contrast ATP decreased binding and decreased basal cGMP accumulation without affecting the ability of ANF to stimulate cGMP. These data indicate that ANF binding and stimulation of cGMP accumulation can be regulated independently supporting further the concept of receptor heterogeneity in renal glomerular membranes. PMID- 2570338 TI - Involvement of platelet-activating factor (PAF) in endotoxin-induced intestinal motor disturbances in rats. AB - Intestinal myoelectrical activity was investigated in conscious fasted rats chronically implanted with Nichrome electrodes in the duodeno-jejunum. Motility of the small intestine was characterized by the presence of migrating myoelectric complex (MMC) occurring regularly at 16.2 +/- 5.8 minute intervals. Intravenous administration of endotoxin (E. coli S.0111:B4) at a dose of 50 micrograms/kg increased the interval between MMC to 112.6 +/- 26.8 min, the duration of these effects being dose-related between 10 to 100 micrograms/kg. Such a typical myoelectrical alteration, corresponding to rapidly propagated groups of spike bursts, was mimicked by the IP administration of PAF at doses of 10 to 50 micrograms/kg. Previous administration of BN 52021, a specific PAF antagonist at a dose of 50 mg/kg abolished the motor alterations induced by IP injection of PAF (25 micrograms/kg) and significantly (p less than 0.01) reduced by 61.2% those induced by IV endotoxin (50 micrograms/kg). Indomethacin (10 mg/kg IP) as well as SC 19220 (5 mg/kg IV), a PGE2 antagonist, injected prior to endotoxin (50 micrograms/kg IV) or PAF (25 micrograms/kg IP) also reduced significantly (p less than 0.01) the duration of MMC inhibition. It is concluded that endogenous release of PAF is partly responsible for the intestinal motor alterations induced by endotoxin; these effects, strongly reduced after treatment with BN 52021, are also mediated through the release of prostaglandins. PMID- 2570339 TI - Temporal analysis of increases in c-fos, preprodynorphin and preproenkephalin mRNAs in rat spinal cord. AB - Peripheral inflammation produces a rapid elevation (within 4 h) in preprodynorphin mRNA in neurons of the dorsal spinal cord and an even more rapid elevation in c-fos proto-oncogene mRNA (within 30 min). During this period a relatively modest increase is also observed in spinal cord preproenkephalin mRNA. Previous anatomical studies have shown that the neurons in which these transcripts increase have overlapping distributions. Assuming that these events occur in the same cells, it suggests the possibility that newly synthesized c-fos protein may participate in transcriptional regulation of opioid genes in spinal cord. PMID- 2570340 TI - Adrenergic receptors mediate changes in c-fos mRNA levels in brain. AB - Recent evidence supports a role for transient c-fos expression as one step in signalling pathways by which membrane receptor-ligand interactions are transduced into appropriate intracellular responses. The activity of adrenergic receptors is mediated by second messenger systems which include ion fluxes, changes in cAMP concentration and enhanced phosphoinositide turnover. In order to determine if C fos induction was also a step in adrenergic signal transduction in the brain, we performed in vivo studies with drugs specific for different adrenergic receptor types. Unexpectedly, we found that the stress associated with a single intraperitoneal (i.p.) injection of drug vehicle produced a transient increase (averaging 4.0-fold) in c-fos mRNA levels in rat brain. Injection of the alpha 2 adrenoreceptor antagonist, yohimbine, produced a transient increase which was larger in magnitude (averaging 9.6-fold) and longer in duration than that produced by injection of the drug vehicle alone. In experiments designed to ask whether either of these inductions was mediated by specific types of adrenergic receptors, we found that the alpha 2- and beta-adrenoreceptors were involved in both responses, while the alpha 1-receptor played a role in mediating the yohimbine induction, but no detectable role in the solvent induction. One hypothesis consistent with our results is that the norepinephrine (NE) released due to the stress associated with an i.p. injection interacts with postsynaptic beta-adrenergic receptors, resulting in the observed c-fos mRNA induction. When the negative feedback effect of NE, mediated by presynaptic alpha 2-receptors, is blocked by yohimbine, the postsynaptic response is enhanced and prolonged. PMID- 2570341 TI - Developmental induction of glutaminase in primary cultures of cerebellar granule cells. AB - Glutaminase mRNA levels increased over 3-fold relative to total RNA, poly(A)+ RNA, and beta-actin mRNA in neonatal rat cerebellar granule cells as the cells differentiated between days 3 and 8 in culture. In contrast, mRNA levels of another glutamate cycle enzyme, glutamine synthetase, remained constant. Glutaminase protein levels increased per cell more than 2-fold between days 3 and 8, and at least 3-fold by day 10 in these cells. The total amount of glutamate per cell increased about 40% during this period. Glutaminase induction paralleled the development of Ca2+-dependent glutamate release, and the formation of neurites, synaptic vesicles, and synapses. The induction of glutaminase in developing granule cells is consistent with a special role for glutaminase in the synthesis of neurotransmitter glutamate. PMID- 2570342 TI - Aging related changes of neurotransmitters in the visual system. AB - Different neurotransmitter systems in the retina and optic nerve can be modified during aging. An increase in GABA and dopamine receptor density in the retina of senescent rats has been demonstrated. Lack of data exists on the aging-related changes in optic nerve neurotransmitters. However, it is possible that neurotransmitter changes in the optic nerve at the lateral geniculate nucleus are similar to those in other areas of central nervous system. PMID- 2570343 TI - Glucagon and hepatic glucose production: modulation by low-dose bradykinin infusion. AB - The effect of a low-dose bradykinin (BK) infusion (30 ng/kg min) on glucagon induced hepatic glucose production and glucose cycling was studied in five normal volunteers. Studies were performed during constant insulin concentration as achieved by simultaneous somatostatin infusion and insulin replacement. In the basal period glucagon was infused at a rate of 0.5 ng/kg min. Then, glucagon infusion rate was increased to 3 ng/kg min to test the response to hyperglucagonemia. In a second set of experiments BK was infused concomitantly with the high dose glucagon. Each subject served as his own control. BK infusion did not prevent the glucagon-induced rise in hepatic glucose production and glucose cycling. However, at a later stage BK accelerated the negative feedback mechanisms activated by glucagon (decrease in hepatic glucose production) significantly. These findings suggest that intravenous BK may interact with mechanisms involved in the down-regulation of hepatic glucagon effects. PMID- 2570345 TI - A fresh start. Report of a one-day workshop on occupational asthma in the aluminium industry. AB - The workshop was successful from a number of viewpoints. It achieved the general goal of a greater understanding of respiratory problems in the aluminium industry. A free exchange of information and attitudes occurred. While the prevalence of asthma was not high compared with other occupational asthmas, it was noteworthy. The industry had embarked on a programme for the development of standard criteria of diagnosis and of the collection of statistics that related to the condition. Some concerns remain. It is to be hoped that further developments will be derived from the workshop, including opportunities for research and for a better understanding of the occupational-health issues and practices in the primary aluminium industry in Australasia. PMID- 2570344 TI - Purification and characterization of the CS2 pili of colonization factor antigen II produced by human enterotoxigenic Escherichia coli. AB - A subtype (CS2) of the colonization factor antigen II (CFA/II) of human enterotoxigenic Escherichia coli (ETEC) was studied. Analysis revealed that CS2 possessing ETEC was predominant among isolates from traveller's diarrhea at Osaka, Japan. TH61 pili produced by a clinical strain (TH61) were purified as a native form to homogeneity by zone electrophoresis and successive column chromatographies on Sepharose 4B and Phenyl-Sepharose CL-4B. It was demonstrated by immunogold staining technique and bacterial agglutination test that antisera against the purified pili of strain TH61 recognized pili of both strain TH61 and strain #C91f, a control strain possessing only CS2 pili. This suggests that TH61 pili purified in this study are CS2 pili. Subunit (pilin) of the purified pili has a molecular weight of about 16,000. Strains bearing CS2 could attach to human jejunal epithelial cells, and this attachment was inhibited by pretreating the enterocytes with purified pili. These indicate that CS2 pili are a factor responsible for attachment of ETEC bearing CS2 to human intestinal cells. PMID- 2570346 TI - Structure of the HOM2 gene of Saccharomyces cerevisiae and regulation of its expression. AB - In Saccharomyces cerevisiae the HOM2 gene encodes aspartic semi-aldehyde dehydrogenase (ASA DH). The synthesis of this enzyme had been shown to be derepressed by growth in the presence of high concentrations of methionine. In the present work we have cloned and sequenced the HOM2 gene and found that the promoter region of this gene bears one copy of the consensus sequence for general control of amino acid synthesis. This prompted us to study the regulation of the expression of the HOM2 gene. We have found that ASA DH is the first reported enzyme of the related threonine and methionine pathway to be regulated by the general control of amino acid synthesis. PMID- 2570347 TI - Characterization of the gene rimK responsible for the addition of glutamic acid residues to the C-terminus of ribosomal protein S6 in Escherichia coli K12. AB - Ribosomal protein S6 of wild-type strains of Escherichia coli contains up to six glutamic acid residues at its C-terminus. The first two residues are encoded by the structural gene for this protein (rpsF) and the rest are added post translationally. Mutants deficient in this modification were isolated and characterized genetically and biochemically. The S6 protein in these mutants appeared to contain only two glutamic acid residues at the C-terminus as expected. The mutated gene was termed rimK and was mapped at 18.7 min between cmlA and aroA. The rimK gene was cloned into a cosmid vector and its nucleotide sequence determined. Analysis of the transcriptional and translational products of this gene indicates that it encodes a protein with an Mr of 31.5 kDa and that it forms an operon with a gene encoding a 24 kDa protein. An rpsF mutant containing a Glu to Lys replacement in the second residue from the C-terminus of protein S6 was isolated. The S6 protein of this mutant was apparently inaccessible to the RimK modification system. This indicates that the RimK modification system requires the wild-type amino acid sequence at least in the C terminal region of ribosomal protein S6. PMID- 2570348 TI - Three regulatory systems control expression of glutamine synthetase in Saccharomyces cerevisiae at the level of transcription. AB - The GLN1 gene of Saccharomyces cerevisiae was cloned by complementation of a gln1 auxotroph. A GLN1-lacZ fusion was constructed to assay GLN1 promoter activity. beta-Galactosidase and glutamine synthetase expression in chromosomally integrated GLN1-lacZ fusion strains were co-regulated in response to a shift from glutamine to glutamate as the nitrogen source, purine limitation, and 3 aminotriazole-induced histidine starvation. Regulation of GLN1 expression by each of the three pathways occurred at the transcriptional level. Increased accumulation of GLN1 mRNA was observed within 5 min after a shift from glutamine to glutamate as the nitrogen source. After 5 min, GLN1 mRNA levels were constant. The level of GLN1 transcript was reduced by approximately 75% within 5 min following glutamine addition to the cells growing with glutamate as nitrogen source. This indicates that the GLN1 message is unstable and has a half-life of approximately 3 min. Deletion analysis indicated that the sequences required for GLN1 expression are located within approximately 350 bp upstream from the transcriptional initiation site. PMID- 2570349 TI - The Klebsiella pneumoniae PII protein (glnB gene product) is not absolutely required for nitrogen regulation and is not involved in NifL-mediated nif gene regulation. AB - The role of the Klebsiella pneumoniae PII protein (encoded by glnB) in nitrogen regulation has been studied using two classes of glnB mutants. In Class I mutants PII appears not to be uridylylated in nitrogen-limiting conditions and in Class II mutants PII is not synthesised. The effects of these mutations on expression from nitrogen-regulated promoters indicate that PII is not absolutely required for nitrogen control. Furthermore the uridylylated form of PII (PII-UMP) plays a significant role in the response to changes in nitrogen status by counteracting the effect of PII on NtrB-mediated dephosphorylation of NtrC. PII is not involved in the nif-specific response to changes in nitrogen status mediated by NifL. PMID- 2570350 TI - Proteins induced by DNA-damaging agents in cultured Drosophila cells. AB - In Drosophila cultured cells, the effects of several DNA-damaging agents on the expression of proteins were investigated. Poly(A+) RNA prepared from both untreated cells and cells treated with DNA-damaging agents was translated in vitro. The translation products were analyzed by two-dimensional electrophoresis. Methyl methanesulfonate, the most potent agent used, induced about 25 proteins, some new and some enhanced pre-existing proteins. Angelicin plus near UV irradiation, 4-nitroquinoline N-oxide and ethyl methanesulfonate were efficient inducers. Mitomycin C, UV irradiation and hydrogen peroxide were poor inducers, inducing only a few proteins at low levels. A tumor promoter, 12-O tetradecanoylphorbol-13-acetate, and a DNA gyrase inhibitor, nalidixic acid, also were used. In this system they were weak inducers of new proteins. Several of the new or enhanced proteins were common to several agents, but others were agent specific. The distribution of mutagen-induced proteins was compared with that of proteins induced in cells heated at 37 degrees C. Some of the proteins induced by DNA-damaging agents were found to overlap heat-shock proteins. These results suggest that there are sets of induced genes that are regulated differently. PMID- 2570352 TI - Necrotizing vasculitis and HIV replication in peripheral nerves. PMID- 2570351 TI - Improved characterization of Theileria parva isolates using the polymerase chain reaction and oligonucleotide probes. AB - Theileria parva DNA was purified from piroplasms isolated from cattle infected with 5 different East African isolates of the parasite, including the East Coast fever reference stock T. p. parva Muguga. Total cellular DNA was prepared from T. parva schizont-infected bovine lymphoblastoid cell cultures (11 isolates). Two probes, previously isolated from T. p. parva Muguga repetitive genomic DNA, were hybridized to restriction digests; closely similar restriction fragment length polymorphism (RFLP) patterns were produced, and no two isolates had the same RFLP pattern. The DNA sequences of probe PMB3, two further copies of the repeated sequence from T. p. parva Muguga, and homologous regions from T. p. parva Kiambu 4 and T. p. lawrencei 3081, were determined. Oligonucleotides were synthesized corresponding to two conserved sections flanking a region which varied between isolates. These oligonucleotides were used as primers in the polymerase chain reaction to amplify the variable region. Further oligonucleotides corresponding to sequences in this variable region were able to distinguish between isolates and no sample hybridized to both oligonucleotides. This is the first unequivocal plus/minus discrimination between groups of isolates to be achieved for T. parva. PMID- 2570353 TI - Synaptic transmission. Broad minded on narrow spikes. PMID- 2570355 TI - Marrow transplantation. Can cord blood be used? PMID- 2570354 TI - Alpha-adrenergic inhibition of sympathetic neurotransmitter release mediated by modulation of N-type calcium-channel gating. AB - In sympathetic neurons, catecholamines interact with prejunctional alpha adrenergic receptors to reduce delivery of transmitter to postjunctional target organs. This autoinhibitory feedback is a general phenomenon seen in diverse neurons containing a variety of transmitters. The underlying mechanisms of alpha adrenergic inhibition are not clear, although decreases in cyclic AMP and cAMP mediated phosphorylation have been implicated. We have studied depolarization induced catecholamine release and calcium-channel currents in frog sympathetic neurons. Here we show that alpha-adrenergic inhibition of transmitter release can be explained by inhibition of Ca2+-channel currents and not by modulation of intracellular proteins. Noradrenaline strongly reduces the activity of N-type Ca2+ channels, the dominant calcium entry pathway triggering sympathetic transmitter release, whereas L-type Ca2+ channels are not significantly inhibited. The down-modulation of N-type channels involves changes in rapid gating kinetics but not in unitary flux. This is the first detailed description of inhibition of a high-voltage activated neuronal Ca2+ channel at the single channel level. The coupling between alpha-adrenergic receptors and N-type channels involves a G protein, but not a readily diffusible cytoplasmic messenger or protein kinase C, and may be well suited for rapid and spatially localized feedback-control of transmitter release. PMID- 2570356 TI - Cyclase enzymes. Deciphering the mosaic. PMID- 2570357 TI - Interaction between peptide growth factors and homoeobox genes in the establishment of antero-posterior polarity in frog embryos. AB - The expression of the Xenopus homoeobox gene xhox3 is an early response to mesoderm induction by peptide growth factors and the level of xhox3 expression marks the antero-posterior character of the induced mesoderm. Different peptide growth factors specify different antero-posterior mesodermal cell fates as seen by the level of xhox3 expression and the capacity to induce specific secondary neural/epidermal structures. These factors and homoeobox genes thus form part of the mechanism necessary for establishing antero-posterior polarity in the frog embryo. PMID- 2570358 TI - Differential activation by atrial and brain natriuretic peptides of two different receptor guanylate cyclases. AB - Alpha atrial natriuretic peptide (alpha-ANP) and brain natriuretic peptide are homologous polypeptide hormones involved in the regulation of fluid and electrolyte homeostasis. These two natriuretic peptides apparently share common receptors and stimulate the intracellular production of cyclic GMP as a second messenger. Molecular cloning has defined two types of natriuretic peptide receptors: the ANP-C receptor of relative molecular mass (Mr) 60-70,000 (60-70 K), which is not coupled to cGMP production and may function in the clearance of ANP and the ANP-A receptor of Mr 120-140 K, which is a membrane form of guanylate cyclase in which ligand binding to the extracellular domain activates the cytoplasmic domain of the enzyme. Here we report the cloning and expression of a second human natriuretic peptide-receptor guanylate cyclase, the ANP-B receptor. The ANP-B receptor is preferentially activated by porcine brain natriuretic peptide rather than human alpha-ANP, whereas the ANP-A receptor responds similarly to both natriuretic peptides. These observations may have important implications for our understanding of the central and peripheral control of cardiovascular homeostasis. PMID- 2570359 TI - The inhibition of alpha 1-adrenoceptor-mediated contractions of rabbit pulmonary artery by Ca2+-withdrawal, pertussis toxin and N-ethylmaleimide is dependent on agonist intrinsic efficacy. AB - Contractions were induced in rings of rabbit pulmonary artery with the preferential alpha 1-adrenoceptor agonists, phenylephrine, methoxamine and St 587 [2-(2-chloro-trifluoromethyl-phenylimino)imidazolidine and the preferential alpha 2-adrenoceptor agonists, clonidine and B-HT 920 [6-allyl-2-amino-5,6,7,8 tetrahydro-4H-thiazolo-(4,5-d) azepine] [corrected]. Phenylephrine and methoxamine acted as full agonists whereas St 587, clonidine and B-HT 920 were partial agonists (intrinsic activities 0.62, 0.38 and 0.42, respectively). Experiments with alpha 1- and alpha 2-adrenoceptor antagonists indicated that the receptors involved are of the alpha 1 type only. Removal of extracellular Ca2+ inhibited maximal contractions to phenylephrine and methoxamine by 30% and 49%, respectively. The remaining contraction components of the full agonists were abolished by the "intracellular Ca2+ antagonist" TMB-8 [8-(N,N-diethylamino)octyl 3,4,5-trimethoxybenzoate]. Contractions to St 587, clonidine and B-HT 920 were virtually abolished in Ca2+-free medium. Pretreatment of the donor rabbits with pertussis toxin (2.5 micrograms/kg i.v., 5-6 days before sacrifice) attenuated the efficacies of the full agonists, phenylephrine and methoxamine by only 24% and 17%, respectively, whereas maximal contractions to the partial agonists, St 587, clonidine and B-HT 920, were inhibited by 46%, 61% and 75%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570360 TI - N-methyl-D-aspartate (NMDA) receptor-mediated stimulation of noradrenaline release, but not release of other neurotransmitters, in the rat brain cortex: receptor location, characterization and desensitization. AB - Rat brain cortex slices and synaptosomes (in a few experiments also hippocampal synaptosomes) preincubated with 3H-noradrenaline, 3H-5-hydroxytryptamine, 3H choline, 3H-glutamate or 3H-gamma-aminobutyric acid were used to investigate the 3H-transmitter release in response to exposure to N-methyl-D-aspartate (NMDA) and other excitatory amino acids. The slices and synaptosomes were superfused with Mg2+-free, otherwise physiologically composed salt solution. In cortical slices preincubated with 3H-noradrenaline, NMDA concentration-dependently stimulated 3H overflow, whereas no such effect occurred in slices preincubated with 3H-5 hydroxytryptamine, 3H-choline, 3H-glutamate or 3H-gamma-aminobutyric acid. In cortical slices preincubated with 3H-noradrenaline, the NMDA-evoked 3H overflow was abolished by tetrodotoxin, presence of Mg2+ 1.2 mmol/l or absence of Ca2+. 2 Amino-5-phosphonovaleric acid produced a parallel shift to the right of the NMDA concentration-response curve, whereas (+)-5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohept- 5,10-imine hydrogen maleate (MK-801) not only shifted the concentration-response curve to the right but also reduced the maximum effect of NMDA. Other excitatory amino acid receptor agonists also stimulated 3H overflow, yielding the following rank order of potency: NMDA greater than L-glutamate greater than L-aspartate. Kainate and, in particular, quisqualate exhibited only low potencies and/or intrinsic activities. Prolonged (25 min) exposure of 3H-NA preincubated cortical slices to a high NMDA concentration produced a short lasting peak of 3H overflow, followed by a second phase lasting as long as the compound was present; in this phase, 3H overflow was clearly less pronounced and gradually decreased with time.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570361 TI - Evidence for selective inhibition of limbic forebrain dopamine synthesis by 8-OH DPAT in the rat. AB - Regional dopamine synthesis in the rat striatum was estimated by measuring DOPA accumulation following inhibition of cerebral aromatic L-amino acid decarboxylase by means of NSD-1015, 100 mg kg-1 intraperitoneally. In animals treated with reserpine, 5 mg kg-1 subcutaneously -18 h, there was a statistically significant increase in DOPA accumulation in the nucleus accumbens, the ventro-medial neostriatum, the dorso-lateral neostriatum and in the posterior limb of the neostriatum. This increase in DOPA accumulation was antagonized dose-dependently in the nucleus accumbens and ventro-medial neostriatum, but not in the other two regions, by treatment with the 5-HT1A receptor agonist 8-OH-DPAT, 0.15-2.4 mumol kg-1, whereas the partial dopamine D2 receptor agonist (-)3-PPP, 2.5-10.0 mumol kg-1, or the full dopamine D2 receptor agonist quinpirole, 0.05-0.8 mumol kg-1, antagonized the reserpine-induced increase in DOPA accumulation uniformly in all four regions of the striatum. The suppression of DOPA accumulation by 8-OH-DPAT in reserpine-treated animals, was completely antagonized by raclopride, 1 mumol kg-1, but not by (-)pindolol, 8 mumol kg-1. The accumulation of 5-HTP in all regions of the striatum as well as in the neocortex following decarboxylase inhibition and reserpine pretreatment, was also inhibited by 8-OH-DPAT, and this inhibition was unaffected by treatment with raclopride or (-)pindolol. It is concluded that 8-OH-DPAT, in addition to general effects on forebrain 5 hydroxytryptamine synthesis, selectively affects limbic forebrain dopamine synthesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570362 TI - Endothelium- and sydnonimine-induced responses of native and cultured aortic smooth muscle cells are not impaired by nitroglycerin tolerance. AB - Tolerance to the cyclic GMP-mediated vasodilator action of nitroglycerin develops with prolonged exposure and may be mediated either by formation of less nitric oxide from nitroglycerin or by desensitization of soluble guanylate cyclase to activation with nitric oxide. In the latter case, smooth muscle cells tolerant to nitroglycerin should show cross-tolerance to nitric oxide released from sydnonimines and endothelial cells (endothelium-derived relaxing factor). Therefore cultured smooth muscle cells from rabbit aorta were pretreated for 1 h with vehicle or high concentrations (0.55 mM) of nitroglycerin or the sydnonimine SIN-1. The formation of cyclic GMP induced by subsequent small doses of nitroglycerin, sydnonimine SIN-1 and endothelium-derived relaxing factor (released from cultured endothelial cells) was compared with the changes in activation of soluble guanylate cyclase, cyclic GMP formation and vasodilation in response to the same stimuli in similarly pretreated segments from rabbit thoracic aortae. Both cultured and native smooth muscle cells remained responsive to stimulation with sydnonimine SIN-1 and endothelium-derived relaxing factor after pretreatment with nitroglycerin, vehicle, or sydnonimine SIN-1, even though they were tolerant to nitroglycerin after pretreatment with nitroglycerin. In contrast, activation of soluble guanylate cyclase by nitroglycerin and sydnonimine SIN-1 was attenuated in homogenates of nitrate-tolerant aortae. The findings suggest that nitroglycerin tolerance in intact cells does not involve desensitization of soluble guanylate cyclase, because in intact cells nitrate tolerance can be overcome by direct activators of soluble guanylate cyclase. PMID- 2570364 TI - Hypothalamic somatostatin may mediate endotoxin-induced fever in the rat. AB - (1) The changes in rectal temperature produced by an injection of a bacterial endotoxin piromen (10-40 ng in 1.0 microliter) on somatostatin-14 (SS-14; 0.1-0.3 microgram in 1.0 microliter) into the preoptic anterior hypothalamic area were assessed and compared in control rats, in rats with hypothalamic SS depletion, and in rats with hypothalamic SS receptor blockade. (2) Intrahypothalamic injection of either piromen or SS-14 produced a dose-related rise in rectal temperature in intact, control rats. The fever induced by intrahypothalamic injection of piromen or SS-14, as well as that induced by intraperitoneal injection of piromen, was antagonized by pretreatment of the hypothalamus with a SS-14 receptor antagonist (0.1 ng in 1.0 microliter) in rats. (3) On the other hand, intraperitoneal administration of cysteamine (30-100 mg/kg), in addition to producing a dose-related fall in rectal temperature, also caused a dose-related fall in hypothalamic SS-levels in rats. Furthermore, the fever induced by intrahypothalamic injection of piromen, but not SS-14, was antagonized by depletion of hypothalamic SS levels with an intraperitoneal dose of cysteamine (30 mg/kg). (4) The results indicate that a somatostatinergic pathway in the hypothalamus may mediate endotoxin-induced fever in the rat. PMID- 2570363 TI - Adenosine inhibits renin release induced by suprarenal-aortic constriction and prostacyclin. AB - The purpose of this study was to determine whether adenosine can attenuate the renin release response to a reduction in renal perfusion pressure. To this end, the secretion rate of renin was measured in six beta-blocked dogs at ambient arterial blood pressure and after a reduction of renal perfusion pressure to 80 mm Hg. These measurements were made during a control period, an intrarenal infusion of adenosine at 10 and 30 micrograms/min, and a recovery period. During the control and recovery periods renal artery hypotension significantly increased the secretion rate of renin. However, during the intrarenal infusions of adenosine, renin secretion rate did not increase significantly. Analysis of variance indicated that both doses of adenosine reduced the renin response to renal artery hypotension. In another six dogs with a single nonfiltering kidney, we again measured renin secretion during a control period, the intrarenal infusion of adenosine at 10 and 30 micrograms/min, and a recovery period; however, in this study PGI2 was used to stimulate renin release. Adenosine also significantly attenuated the renin release response to PGI2. We conclude that adenosine can inhibit the renin release response to both renal artery hypotension and PGI2 and that this effect is most likely mediated by a direct action of adenosine on juxtaglomerular cells. Also, since PGI2 may be a mediator of the renin response to renal artery hypotension, the data are consistent with the hypothesis that adenosine inhibits the renin response to renal artery hypotension by attenuating the response of juxtaglomerular cells to PGI2. PMID- 2570365 TI - Mechanisms of chronotropic cardiac effects of alinidine and plasma concentration response relationships in the conscious dog with chronic atrioventricular block. AB - The chronotropic cardiac effects of alinidine were studied in the conscious dog with chronic atrioventricular block. Alinidine at 0.5 - 4 mg/kg, i.e., at plasma concentrations between 42 +/- 2 and 1625 +/- 371 ng/ml, initially increased atrial rate dose-dependently. This effect fell off rapidly, but atrial bradycardia was never observed. After atropine and pindolol, which raised basal atrial rate, alinidine (2 mg/kg) decreased atrial rate, whereas after phenoxy benzamine, yohimbine or phentolamine, it produced atrial effects identical to those observed under basal conditions, i.e., initial tachycardia and no bradycardia. Alinidine dose-relatedly decreased ventricular rate. None of the pretreatments modified the maximal ventricular bradycardia, but interestingly after pindolol or yohimbine this effect developed more rapidly (maximal bradycardia between 3 and 5 against 30 min) and then declined progressively. Alinidine did not modify mean blood pressure at any dose. After atropine, phenoxybenzamine or phentolamine, alinidine remained without effect on mean blood pressure, but after pindolol or yohimbine, a hypotensive effect appeared concomitantly with the reduction of the ventricular bradycardia. These results show that the initial atrial cardioacceleration due to alinidine results from a direct vagolytic action of this drug and that the absence of atrial bradycardia results from buffering by the vagolytic effect and/or a relatively low basal atrial rate. They also suggest that the ventricular bradycardia does not involve either the muscarinic cholinoceptors or the alpha- or beta-adrenoceptors, though the results obtained after pindolol or yohimbine suggest possible involvement of a fall in sympathetic tone by stimulation of presynaptic or central alpha2 adrenoceptors, particularly in the persistence of the bradycardic effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570368 TI - Methylxanthines augment the renin response to suprarenal-aortic constriction. AB - In a previous study we discovered that the adenosine receptor antagonist, caffeine, increases plasma renin activity and blood pressure in renin-dependent renovascular hypertension. The purpose of the present investigation was to determine whether methylxanthines augment the increase in renin secretion induced by a reduction in renal perfusion pressure and, if so, whether this effect is mediated by a direct action on juxtaglomerular cells. Accordingly, we examined the effects of infusions of caffeine and theophylline directly into the renal artery on the increase in renin secretion induced by suprarenal aortic constriction. All studies were conducted in dogs receiving an intravenous infusion of propranolol to prevent changes in renin secretion mediated indirectly via the sympathetic nervous system. Caffeine (5 mg/min) increased the renin response to suprarenal aortic constriction about 10-fold without significantly affecting renal hemodynamics or excretory function. Theophylline (5 mg/kg), on the other hand, did not significantly increase the renin response to a reduction in renal perfusion pressure, but did increase urine flow and sodium excretion about 10-fold. However, in the non-filtering, beta-adrenoceptor blocked, canine kidney, theophylline markedly increased the renin response to suprarenal aortic constriction. These results indicate that methylxanthines can potentiate the renin response to a reduction in renal perfusion pressure most likely by directly affecting the juxtaglomerular cells; however, since increased sodium delivery to the macula densa inhibits renin release, the extent to which methylxanthines affect the renin response to renal artery hypotension depends on how vigorous the diuretic response is to a given methylxanthine. PMID- 2570367 TI - Influence of cooling on the responsiveness of rat isolated anococcygeus muscle to noradrenaline. AB - The isolated anococcygeus muscle of the rat was used to study the effect of temperature on noradrenaline-induced contraction. The preparation was suspended in an organ bath containing Krebs bicarbonate solution for isometric tension recording. A decrease of the bath temperature from 37 degrees C to 20 degrees C (cooling) produced an increase in tissue sensitivity to noradrenaline, as reflected in a 5.37-fold leftward shift in the concentration-response curve, and increased the maximum contractile response to this agonist (14.3%). Cooling had no effect on tissue sensitivity to a selective alpha 1-adrenoceptor agonist, methoxamine, but increased (12.4%) the maximum contraction to a similar extent to that to noradrenaline. 6-Hydroxydopamine pretreatment or nortriptyline (1 mumol/l) induced a leftward shift of the noradrenaline concentration-response curve at 37 degrees C, and profoundly inhibited the potentiating effect of cooling on tissue sensitivity to the catecholamine; the effect of cooling on the maximum response was unaffected. The affinity of noradrenaline or methoxamine for alpha 1-adrenoceptors at 37 degrees C, determined from its dissociation constant (KA), was not significantly different from that at 20 degrees C. KA values were determined by use of irreversible antagonism with phenoxybenzamine. On the other hand, diltiazem at concentration of 3 mumol/l, which almost completely abolished the calcium ion-induced contraction of the potassium ion-depolarized muscle, caused only slight inhibition in the concentration-response curve for noradrenaline. The contractile responses to Ca2+ of the K+-depolarized muscle and of the tissue incubated in Ca2+ -free (EGTA 0.1 mmol/l) Krebs solution containing diltiazem and noradrenaline were both depressed by cooling.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570369 TI - Mass and in situ molar activity of tyrosine hydroxylase in the median eminence. Effect of thyroidectomy and thyroid hormone replacement. AB - The effects of thyroidectomy and thyroid hormone replacement on the mass and in situ molar activity of tyrosine hydroxylase (TH) in the median eminence (ME) and superior cervical ganglia (SCG) of male rats were investigated. The tissue specificity of these effects were evaluated by comparing the ME with the superior cervical ganglion (SCG). All animals were thyroparathyroidectomized (Tx) or sham Tx. Tx rats were treated daily for 3 weeks with 0.15 M NaCl (solvent vehicle) or L-thyroxine (T4). Two doses of T4, 10 and 100 micrograms/day/kg BW, were used. Sham Tx rats were treated with 0.15 M NaCl. All animals were studied on the day following the last treatment. The mass of TH was determined using an immunoblot assay, and the in situ activity of TH was calculated from the rate of intracellular accumulation of L-dihydroxyphenylalanine (DOPA) after administration of an inhibitor of DOPA decarboxylase activity. In the ME, thyro parathyroidectomy resulted in a 40% increase in the mass and a 100% increase in the in situ molar activity of TH over that of sham Tx rats. Compared to Tx animals given 0.15 M NaCl, Tx rats treated with a low dose of T4 (10 micrograms/day/kg BW) had a reduced quantity of TH in the ME, but the molar activity of the enzyme was increased. Treatment of Tx rats with a high dose of T4 (100 micrograms/day/kg BW) restored TH mass but not the in situ activity of TH in the ME to the level seen in sham Tx rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570370 TI - Hydrocephalus-induced changes in the composition of cerebrospinal fluid. AB - Studies reporting the composition of cerebrospinal fluid obtained from hydrocephalic humans have been critically reviewed. Hydrocephalus-induced alterations in the cerebrospinal fluid concentrations of neurotransmitters and peptide neuromodulators, products of glycolysis and nucleotide metabolism, neural cell-derived proteins and enzymes, and serum-derived proteins have been documented. The data are interpreted with reference to experimental studies. The reported changes suggest that in the hydrocephalic brain there are disturbances of oxidative metabolism and neurotransmission, and perhaps damage to periventricular cells particularly when intracranial pressure is elevated. Although no assays have provided and entirely useful guide to aid decisions regarding shunt therapy, they have provided in vivo information regarding the pathophysiology of hydrocephalus. PMID- 2570366 TI - Veratridine-induced phosphorylation and activation of tyrosine hydroxylase, and synthesis of catecholamines in cultured bovine adrenal medullary cells. AB - The mechanism of the synthesis of catecholamines by veratridine was studied in cultured bovine adrenal medullary cells. (1) Veratridine increased the phosphorylation and activity of tyrosine hydroxylase as well as the synthesis of [14C]catecholamines from [14C]tyrosine, all of which were inhibited by tetrodotoxin. Veratridine-induced activation of tyrosine hydroxylase and synthesis of [14C]catecholamines were reduced in 20 mmol/l extracellular Na+ or in Ca2+-free medium. (2) 12-O-Tetradecanoylphorbol-13-acetate (TPA), an activator of protein kinase C, increased the synthesis of [14C]catecholamines. In the presence of TPA, veratridine did not produce any additional increase in [14C]catecholamine synthesis. In protein kinase C-deficient cells which were prepared by pretreatment with 1 mumol/l TPA for 24 h, TPA failed to increase [14C]catecholamine synthesis and veratridine-induced [14C]catecholamine synthesis was suppressed by 50%. (3) Polymyxin B, an inhibitor of protein kinase C and N-(6 aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7), an inhibitor of calmodulin, inhibited veratridine-stimulated synthesis of [14C]catecholamines as well as veratridine-induced influx of 22Na+ and 45Ca2+ with similar potencies. (4) In digitonin-permeabilized cells, polymyxin B attenuated the activation of tyrosine hydroxylase caused by Ca2+. These results suggest that veratridine-induced synthesis of catecholamines and activation of tyrosine hydroxylase were mediated by Ca2+-dependent phosphorylation of this enzyme, and protein kinase C may be responsible, at least in part, for this process. PMID- 2570371 TI - Fast hyperpolarization following an excitatory postsynaptic potential in cat bladder parasympathetic neurons. AB - Intracellular recording techniques were used to study a fast hyperpolarizing potential following the fast excitatory postsynaptic potential evoked by an orthodromic nerve stimulation in cat bladder parasympathetic ganglion cells. In the 61 ganglion cells examined, two types of responses were recorded on stimulating the preganglionic nerve; one had only a fast excitatory postsynaptic potential (type SI, n = 20) and the other had a fast excitatory postsynaptic potential followed by a fast hyperpolarizing potential (type SII, n = 41). In type SII neurons, the half-maximum duration of the afterhyperpolarizing potential following an orthodromic spike was longer than that of a direct spike produced by injecting a depolarizing current pulse through the recording electrode; the half maximum durations for afterhyperpolarizing potentials following orthodromic and direct action potentials were comparable in type SI cells. Blocking the initiation of an orthodromic spike by hyperpolarizing the membrane in type SII cells revealed a fast excitatory postsynaptic potential followed by a fast hyperpolarizing potential which was similar to that observed at the resting potential. The fast hyperpolarizing potential had a duration comparable to that of an afterhyperpolarizing potential following an orthodromic action potential. The fast excitatory postsynaptic potential-fast hyperpolarizing potential sequence was blocked completely and reversibly by nicotinic receptor antagonists (hexamethonium and D-tubocurarine). Atropine, alpha-2 noradrenergic (yohimbine and phentolamine), and purinergic (caffeine) antagonists had no effect on the fast hyperpolarizing potential. In cells which show type SII responses, spontaneous excitatory postsynaptic potentials were not followed by a hyperpolarization. Depolarizing the membrane (by passing a cathodal current through the recording electrode) to an amplitude comparable to that of a fast excitatory postsynaptic potential also did not elicit a membrane hyperpolarization in type SII cells. In some cells, stimulating one preganglionic nerve trunk elicited a fast hyperpolarizing potential, but activating another nerve trunk innervating the same ganglion cell did not. There was no correlation between the variations in the amplitudes of the fast excitatory postsynaptic potential and the fast hyperpolarizing potential in type SII cells, but increasing the stimulus intensity applied to the presynaptic nerve fiber potentiated the amplitude of the fast excitatory postsynaptic potential and the fast hyperpolarizing potential. The fast hyperpolarizing potential was not associated with appreciable changes in input resistance.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2570372 TI - Nordihydroguaiaretic acid blocks the synaptic component of long-term potentiation and the associated increases in release of glutamate and arachidonate: an in vivo study in the dentate gyrus of the rat. AB - The dentate gyrus of anaesthetized rats was perfused with artificial cerebrospinal fluid while field responses evoked by stimulation of the perforant path were monitored. Perfusates were collected for analysis of endogenous glutamate, aspartate and arachidonate. In animals in which long-term potentiation was induced by tetanic stimulation, there was a sustained increase in the concentration of glutamate in the perfusate, and, less reliably, in aspartate, as previously reported by Bliss et al. (J. Physiol., Lond. 377, 391-408, 1986) and Errington et al. (Neuroscience 20, 279-284, 1987). The lipoxygenase and phospholipase A2 inhibitor nordihydroguaiaretic acid, when added to the perfusate 30 min before the tetanus, abolished both long-term potentiation of the population excitatory postsynaptic potential and the tetanus-induced increase in glutamate release. Long-term potentiation of the population spike was reduced but not abolished. There was also a sustained increase in the release of arachidonic acid following the induction of long-term potentiation which did not occur when induction was blocked by nordihydroguaiaretic acid. These results are discussed in the light of the possibility that arachidonic acid or one of its lipoxygenase metabolites may be the retrograde messenger which we have postulated is released from postsynaptic sites following tetanic stimulation to trigger increased transmitter release from presynaptic terminals. PMID- 2570373 TI - Phosphorylation of tyrosine hydroxylase in protein kinase C-deficient PC12 cells. AB - In order to study the role of protein kinase C in the regulation of tyrosine hydroxylase phosphorylation in PC12 cells, the effects of various agonists on diacylglycerol accumulation in PC12 cells were measured and the ability of these agonists to increase the phosphorylation tyrosine hydroxylase in protein kinase C deficient cells was evaluated. Bradykinin (10 microM) and elevated extracellular K+ (55 mM) increased the accumulation of [3H]diacylglycerol in PC12 cells that had been prelabeled with [3H]arachidonic acid, and so might be expected to activate protein kinase C in these cells; in contrast, nerve growth factor did not increase diacylglycerol accumulation in PC12 cells. Protein kinase C deficient PC12 cells were prepared by incubating the cells for 24 h with 1 microM phorbol dibutyrate. This treatment resulted in the loss of approximately 90% of the protein kinase C activity in the cells. Control and protein kinase C deficient cells were incubated with 32Pi for 90 min and then stimulated with various agonists. 32P-labeled tyrosine hydroxylase was isolated from the cells by polyacrylamide gel electrophoresis and subjected to tryptic hydrolysis. 32P containing phosphopeptides were separated by two-dimensional thin-layer electrophoresis and chromatography, visualized by autoradiography, and quantitated by scintillation counting Treatment of control cells with phorbol dibutyrate increased the incorporation of 32P into one tryptic phosphopeptide (referred to as T3) in tyrosine hydroxylase. Phorbol dibutyrate did not increase the phosphorylation of this peptide in protein kinase C-deficient cells. Bradykinin or 55 mM K+ increased the incorporation of 32P into four tyrosine hydroxylase phosphopeptides, including peptide T3.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570374 TI - Sennosides do not kill myenteric neurons in the colon of the rat or mouse. AB - Effects of senna on the myenteric plexus of the colon were investigated in view of earlier reports that this anthraquinone cathartic depletes the plexus of its intrinsic neurons. Rats and mice were given purgative doses of sennosides in their drinking water for 4 and 5 months, respectively. Body growth was reduced, and the weight of the colon with its contents was increased relative to the weight of the whole body in the treated animals. The latter change was attributed to depressed propulsive motility of the large intestine. Total numbers of myenteric neurons were determined from whole-mount preparations stained with Cuprolinic Blue-magnesium chloride, which selectively coloured the neuronal somata. The number of neurons in the rat's colon was unaffected by treatment with senna, but the colons of the treated mice contained significantly more neurons than those of their controls. Staining with antisera to 10 putative neurotransmitters or their associated enzymes revealed immunoreactive somata and axons in the myenteric plexus. Treatment with senna was not associated with absence of neuronal somata or fibres stainable with any of the antisera in either species. Thus, there was no evidence of toxic destruction of any identifiable population of neurons that might have been too small to affect the total counts. We conclude that senna does not kill myenteric neurons in the colon of the rat or mouse. PMID- 2570375 TI - Biochemical properties of brain somatostatin receptors. AB - The physical properties of brain and pituitary somatostatin receptors were characterized using photocrosslinking techniques. Somatostatin receptors in rat corpus striatum and anterior pituitary membranes were covalently bound to the non reducible somatostatin analog, [125I]CGP 23996, using the crosslinking agent n hydroxysuccinimidyl-4-azidobenzoate and ultraviolet light. In striatal membranes, a protein of 60,000 mol. wt was labeled by [125I]CGP 23996. The binding was potently inhibited by somatostatin analogs but not by other biologically active peptides. The labeling of the 60,000 mol. wt protein by [125I]CGP 23996 was diminished by guanine triphosphate gamma thiol, which is consistent with the labeling of a somatostatin receptor coupled to guanine triphosphate binding proteins. The migration of the [125I]CGP 23996 labeled 60,000 mol. wt protein in native sodium dodecyl sulfate-gels was not affected by the reducing agent dithiothreitol, indicating that there is a general lack of disulfide bridges in the striatal somatostatin receptor. The striatal somatostatin receptor was solubilized with the detergent 3-[(3-cholamidopropyl)-dimethylaminoio]-1 propanesulfonate and specifically bound to the lectin wheat germ agglutinin, suggesting that the striatal somatostatin receptor is a glycoprotein. [125I]CGP 23996 also labeled a 60,000 mol. wt protein in anterior pituitary membranes. The characteristics of [125I]CGP 23996 binding to anterior pituitary membranes were consistent with the labeling of a somatostatin receptor. Interestingly, a comparison of the [125I]CGP 23996 labeled material from striatal and anterior pituitary membranes by two-dimensional polyacrylamide gel electrophoresis revealed the presence of several striatal somatostatin receptors of varying charge (pI values between 6 and 6.5) but only a single pituitary receptor. These findings indicate that physical differences may exist between subtypes of somatostatin receptors. PMID- 2570376 TI - Activation of ganglionic tyrosine hydroxylase by peptides of the secretin glucagon family: structure-function studies. AB - The hydroxylation of tyrosine to dopa is the rate-limiting reaction in catecholamine biosynthesis. It has been previously reported that secretin, vasoactive intestinal peptide and peptide histidine isoleucine amide, all members of the secretin-glucagon family of peptides, increase dopa synthesis in superior cervical ganglia in vitro. We report here that two other members of this peptide family, rat growth hormone-releasing factor and helodermin H38, a component of Gila monster venom, also increase the rate of dopa synthesis, while glucagon-like peptides I and II and a number of other peptides tested produce no effect. Since analogs of cAMP also increase dopa synthesis, it is of particular interest that all of the peptides that increase catechol synthesis also raise the levels of this cyclic nucleotide in the superior cervical ganglion. Helodermin H38 stimulated the rate of dopa synthesis and the level of cAMP with similar potencies (EC50S of approximately 10 nM) and with maximal effects of two- and two fold, respectively. By either measure, rat growth hormone-releasing factor produced a two-fold increase at 10 microM and a three- to four-fold increase at 30 microM. Analogs of peptides of the secretin-glucagon family with a deletion or modification of the N-terminal histidine were much less effective in these assays at the concentrations tested than were their parent compounds, demonstrating an important role for this amino acid in conferring activity on these peptides. In addition to increasing dopa synthesis in intact tissue, incubation of ganglia with rat growth hormone-releasing factor, secretin, vasoactive intestinal peptide or peptide histidine isoleucine amide also increased the activity of tyrosine hydroxylase measured subsequently in ganglion homogenates. Thus, the peptidergic stimulation of dopa synthesis observed in the intact superior cervical ganglion appears to be due, at least in part, to the activation of tyrosine hydroxylase. Together with previous studies, these findings support the hypothesis that certain members of the secretin-glucagon family increase catecholamine synthesis in sympathetic neurons by a cAMP-dependent activation of tyrosine hydroxylase. PMID- 2570378 TI - Presynaptic auto- and allelo-receptor regulation of hypothalamic opioid peptide release. AB - Recent studies have shown that inhibitory feedback mechanisms regulate the release of the endogenous opioid peptides beta-endorphin (acting predominantly at mu opioid receptors in the brain), dynorphin (a kappa opioid receptor ligand) and [Met]enkephalin (a delta opioid receptor ligand) from the rat hypothalamus. By using specific antagonists of the various opioid receptor types, it is shown that the release of these peptides from hypothalamic slices in vitro is not only controlled by homologous (auto)-receptors, but that cross-regulation between the three neuronal opioid receptor types also occurs; thus, the delta receptor antagonist N,N-diallyl-Tyr-Aib-Aib-Phe-Leu increases the release of all three peptides, the mu receptor antagonist D-tetrahydroisoquinoline-Cys-Tyr-D-Trp-Arg Thr-Pen-Thr-NH2 increases that of beta-endorphin and dynorphin, and the kappa receptor antagonist nor-binaltorphimine increases that of dynorphin; all these effects occur in the presence of tetrodotoxin, indicating a presynaptic site of action. We propose the term "allelo-receptors" to describe this particular form of neuronal regulation in which an endogenous ligand, acting via its own specific receptor, also regulates the release of related peptides which activate different classes of opioid receptors. PMID- 2570377 TI - Muscarinic excitation and inhibition of neurons in the submucous plexus of the guinea-pig caecum. AB - Intracellular recordings were made from neurons in the submucous plexus of the guinea-pig caecum. Muscarinic agonists (acetylcholine, bethanechol and muscarine) depolarized about 70%, and hyperpolarized about 30% of the submucous plexus neurons. Low concentrations of pirenzepine reversibly antagonized both responses. The measured dissociation constants (KD) of 10-30 nM for the depolarizations and 1-3 nM for the hyperpolarizations suggest that each response was mediated by muscarinic M1 cholinoceptors. The muscarinic depolarization and hyperpolarization were associated with a decreased and an increased conductance, respectively, and the reversal potential for the muscarinic responses varied as the potassium concentration varied, always being around the potassium equilibrium potential. In cells depolarized by muscarinic agonists these agents appeared to decrease a potassium conductance that could also be inactivated by substance P. In approximately 30% of the submucous neurons, the slow inhibitory postsynaptic potential, elicited in response to single or repetitive focal stimuli (1-10 pulses at 20-40 Hz), appeared to consist of a large component which was sensitive to the blocking action of idazoxan (100-300 nM) and a small component which was idazoxan-insensitive. The latter (muscarinic slow inhibitory postsynaptic potential) was completely abolished by pirenzepine. The concentrations of pirenzepine which caused a 50% depression ranged from 5 to 20 nM. The muscarinic slow inhibitory postsynaptic potential was increased in amplitude and duration by physostigmine (100-300 nM). The muscarinic slow inhibitory postsynaptic potential was accompanied by a decrease in membrane input resistance, and was reversed in polarity near the potassium equilibrium potential. When muscarine induced a hyperpolarization and/or focal stimulation elicited a muscarinic slow inhibitory postsynaptic potential in the presence of idazoxan (100-300 nM), the intracellular injection of guanosine 5'-O-(3-thiotriphosphate) produced a progressive membrane hyperpolarization during which the muscarinic hyperpolarizing responses were attenuated. It is concluded that the muscarine induced reduction in potassium conductance is mediated through a muscarinic M1 receptor which has a relatively low affinity for pirenzepine. The muscarine induced increase in potassium conductance is probably produced by the association of a guanine nucleotide-binding regulatory protein with another muscarinic M1 receptor that has a relatively high affinity for pirenzepine. PMID- 2570379 TI - Induction of the gene encoding pro-dynorphin by experimentally induced arthritis enhances staining for dynorphin in the spinal cord of rats. AB - The response of dynorphinergic neurons in the lumbosacral spinal cord of the rat to chronic arthritic inflammation was studied by the combined use of biochemical and immunohistochemical procedures. In polyarthritic rats, in which all four limbs showed a swelling, inflammation and hyperalgesia, a pronounced elevation was seen in the level of messenger ribonucleic acid encoding prodynorphin (pro enkephalin B) in the lumbosacral spinal cord. In addition, the levels of immunoreactive dynorphin A1-17, a primary gene product of this precursor, were greatly increased. This activation was reflected in a striking intensification of the immunohistochemical staining of both dynorphin and alpha/beta-neo-endorphin, a further major product of pro-dynorphin. In control animals perikarya were stained exceedingly rarely and encountered only in laminae I and II. Stained fibres and varicosities were seen throughout the dorsal and ventral gray matter, being most concentrated in laminae I, II, IV and V of the dorsal horn and dorsolateral to the central canal. In polyarthritic rats, fibres and varicosities were much more intensely stained throughout the cord, particularly in laminae I/II, IV and V and dorsolateral to the central canal. Many strongly-stained perikarya could be seen: these comprised many small diameter cells in laminae I and II, and some large diameter marginal neurons and large diameter cells, heterogenous in appearance, in the deeper laminae IV and V. Monolaterally inflamed rats injected in the right hind-paw showed pathological changes only in this limb. Correspondingly, in unilateral inflammation, an elevation in immunoreactive dynorphin was seen exclusively in the right dorsal horn and the above-described intensification of staining for dynorphin and neo-endorphin was seen only in this quadrant. This reveals the neuroanatomical specificity of the response. Thus, in the lumbosacral cord of the rat, pro-dynorphin neurons are most preponderant in laminae I, II, IV and V. A pronounced intensification of the immunohistochemical staining of these neurons is seen in chronic arthritis. Furthermore, there is a parallel elevation in the levels of messenger ribonucleic acid encoding pro-dynorphin and of its primary products dynorphin and neo endorphin. These findings demonstrate an enhancement in the functional activity of spinal cord localized dynorphin neurons in the response to chronic arthritic inflammation. PMID- 2570380 TI - Induction of parkinsonism by intraventricular bethanechol in a patient with Alzheimer's disease. PMID- 2570381 TI - [Anomalies in gastric stump emptying of patients who have undergone resections for duodenal ulcer previously treated with histamine H2 receptor antagonists]. AB - The problem of obstructed emptying of the gastric stump after Billroth II operations in patients previously treated with H2 receptors is discussed. Statistical comparison using the Student's "t" test revealed a significant difference (t = 2.173) between those given and not given H2-antagonists and confirmed the existence of greater postoperative hypotonia in the gastric stumps of the former group. This clinical syndrome demands careful monitoring of the hydroelectrolytic balance and possibly the use of a double naso-gastric and nasojejunal tube as a precaution to ensure the delivery of the jejunal contents to the stomach. The gastrokinetic drugs metoclopramide and domperidone are also beneficial. PMID- 2570383 TI - [Long-term evaluation of the treatment of perforated gastroduodenal ulcer using a median suture]. AB - Simple suture associated with treatment using H2-antagonists has replaced gastroduodenal resection in the treatment of peptic ulcer. Forty-four patients were submitted to rafia suture for perforated gastric or duodenal ulcer (1972 1986). Fourteen were treated in the pre-H2-antagonist period and 30 in the post H2 antagonist period. Patients were followed up: Vissick classes I and II represent 58.3% of cases in the pre-H2-antagonist period and 95.8% in the post-H2 antagonist period. Two patients from the first period were subjected to gastroduodenal resection. In no patient of the second period was further ulcer therapy necessary. In patients of the first period, oesophagogastroduodenoscopy evidenced duodenal ulcer in one case and erosive duodenitis in two cases; erosive duodenitis was present in one patient of the second period. The choice of rafia would appear to be a valid one. Follow-up after a longer period should provide confirmation. PMID- 2570382 TI - [Current approach to the treatment of acute pancreatitis with reference to the use of somatostatin]. AB - A series of 172 cases of acute pancreatitis encountered between 1-1-79 and 30-4 88, including 57 treated with somatostatin is presented. A comparison between the latter and the other cases treated with a variety of drugs (aprotinin, cimetidine, ranitidine) led to the following conclusions: 1) somatostatin significantly improves the clinical course of acute oedematous pancreatitis with circumscribed necrosis; 2) it makes no difference to the development of cases with diffuse necrosis and haemorrhage. PMID- 2570384 TI - Loss of glycine-dependent radioligand binding to the N-methyl-D-aspartate phencyclidine receptor complex in patients with Alzheimer's disease. AB - Well washed membranes have been prepared from samples of cerebral cortex of control subjects and patients with Alzheimer's disease, obtained both at post mortem and by neurosurgical procedures earlier in the course of the disease. Binding to these membranes of two radioligands for the N-methyl-D-aspartate phencyclidine receptor complex has been determined in the presence and absence of glycine. Glycine increased the binding in both control and Alzheimer tissue samples. At one concentration of radioligand, in the presence of glycine there was less binding to post-mortem samples, which Scatchard analysis showed was associated with a 36% loss of sites. In rare neurosurgical samples, there was also a loss of binding of radioligand which suggests that the effect is not due to post-mortem artefacts or epiphenomena. These new results may have implications for the symptomatic and preventative treatment of Alzheimer's disease. PMID- 2570385 TI - MPTP-induced ventral mesencephalic cell loss in the cat. AB - Immunohistochemical analysis of tyrosine hydroxylase (TH)-containing neurons in the cat ventral mesencephalon revealed a marked loss of substantia nigra pars compacta neurons and moderate losses of TH-positive neurons in the ventral tegmental area, retrorubral or A8 region, and in the substantia nigra pars lateralis of cats treated with the neurotoxin N-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). The pattern and magnitude of cell loss in these ventral mesencephalic regions parallels that previously reported for the MPTP treated monkey. These results further support the use of the cat as a model for studying the pathology to catecholaminergic neurons induced by MPTP. PMID- 2570386 TI - Cellular localisation of somatostatin mRNA and neuropeptide Y mRNA in foetal striatal tissue grafts. AB - Using in situ nucleic acid hybridisation histochemistry, we have studied the expression of somatostatin mRNA and neuropeptide Y mRNA in grafts of embryonic striatal neurones implanted into the ibotenic acid-lesioned rat neostriatum. Tissue sections of the grafted striatum were incubated with either 32P- or 35S labelled complementary oligodeoxyribonucleotide probes specific for somatostatin mRNA and neuropeptide Y mRNA, exposed with X-ray film and dipped in Ilford K-5 emulsion. Neither somatostatin mRNA nor neuropeptide Y mRNA was detectable in the ibotenic acid-lesioned striatum indicating a pronounced degeneration of somatostatin- and neuropeptide Y-containing neurones. However, in the striatal grafts the levels of somatostatin mRNA and neuropeptide Y mRNA were substantially increased over those in the control intact striata. The results suggest that in the grafts, somatostatin mRNA and neuropeptide Y mRNA were expressed in a higher proportion of primordial striatal neurones and there was also an increased level of expression of each neuropeptide gene per individual neurone (reflecting a higher synthetic activity of such neurones) compared to the intact mature striatum. These data demonstrate the sensitivity of the in situ hybridisation technique to study patterns of gene expression in developing neuronal tissues after transplantation. PMID- 2570388 TI - Cystine neurotoxicity is increased by taurine deficiency. AB - In an attempt to increase taurine biosynthesis in cats fed a taurine-free diet we supplied an excess of the precursor, cystine, in the diet. All nine cats exhibited extreme signs of neurotoxicity including lethargy, inability to stand, rigidity of the neck and lower limbs, absence and epileptic seizures, severe retinal damage and death. In a similar group of cats fed 0.05% taurine in addition to an excess of cystine, four cats died after showing minimal symptoms of lethargy and unsteadiness and the remainder showed no adverse effects. Biochemical measurements, tissue concentrations of cystine, cysteine, bound cysteine, glutamate and taurine and activities of enzymes involved in taurine biosynthesis, revealed significant differences only in taurine concentrations. PMID- 2570387 TI - Hundred-fold increase in neuronal vulnerability to glutamate toxicity in astrocyte-poor cultures of rat cerebral cortex. AB - In cultures of rat cerebral cortex in which astrocyte proliferation was stringently suppressed, glutamate neurotoxicity occurred at glutamate concentrations similar to those which are normally found in the extracellular space in the hippocampus. Concentrations of glutamate one hundred-fold higher were required to produce neurotoxicity in the presence of abundant astrocytes. This suggests that the sensitivity of central neurons to glutamate toxicity may be dependent upon astrocyte function. PMID- 2570389 TI - Cell specific enzyme markers as indicators of neurotoxicity: effects of acute exposure to methylmercury. AB - In order to assess the sensitivity of several cell specific enzyme markers (tyrosine hydroxylase (TH), glutamic acid decarboxylase, choline acetyltransferase, glutamine synthetase (GS), neuron specific and non-neuronal enolase and 2',3'-cyclic nucleotide phosphohydrolase (CNP] as indices of neurotoxicity, changes in their activities were monitored after rats were treated with two doses of the neurotoxic agent, methylmercury chloride (MMC). Comparisons were also made of any histopathological changes occurring in the tissues examined. At the low dose rate (3.36 mg Hg/kg, po, for 14 days), the rats exhibited less body weight gain compared to untreated animals. No change in either the neuronal or noneuronal enzyme markers was observed in brain but a significant increase in the myelin marker, CNP, and total enolase activity was seen in the optic nerve. Morphological evaluation by light microscopy indicated no discernible neuronal lesions in MMC-exposed animals. At the higher MMC dose (7.05 mg Hg/kg, po, for 7 days), there was about a 20% loss in the body weight of treated animals and partial hind limb paralysis was observed. Of all the neuronal marker enzymes examined, only TH was found to be decreased in the striatum. The proliferating astroglial marker, GS, was elevated only in the cerebellum. CNP was found to be decreased in both the optic and sciatic nerve. As in the lower dose group no pathological changes were observed at the light microscopic level in the brain of MMC-treated rats. These data suggest that of the cell specific marker enzymes studied, GS in the cerebellum and TH in the striatum may be useful biochemical markers for the neurotoxic action of MMC. PMID- 2570390 TI - [Changes in the expression of the surface antigens of thymocytes during their cultivation with macrophages]. AB - The data on changes in expression of H-2 complex and Thy-1 antigens on cell surface of thymocytes resulting from their incubation with peritoneal macrophages has been presented. The process of joint cultivation of thymocytes with macrophages leads to significant decrease in number of cells with Thy-2-antigen and increase in that with H-2 complex antigens. An increase in H-2K+ cells in experimental thymocytes as compared to control ones was observed. No changes in H 2D expression was observed. A significant increase in Ia+ macrophages was observed after interaction with thymocytes as compared with intact mononuclear phagocytes. PMID- 2570392 TI - Factor XIIIa-containing cells and fibrosis in oral and maxillofacial lesions: an immunohistochemical study. AB - The distribution of subunit A of blood coagulation factor XIII (FXIIIa) was investigated by the avidin-biotin-peroxidase complex (ABC) method in various oral and maxillofacial tissues. These tissues were from normal tongue, gingiva, lip, and submandibular gland, and from Dilantin gingival hyperplasia (one case), pyogenic granuloma (three cases), peripheral fibroma (four cases), squamous cell carcinoma (seven cases), chronic sclerosing submandibular adenitis (two cases), and fibrous dysplasia of the mandibular bone (one case). The distribution of collagenous components was examined in the same tissues by means of the Sirius red F3BA method. By means of the ABC method, FXIIIa was detected in the cytoplasm of certain connective tissue cells in each of the tissues examined. These FXIIIa containing cells were sparse in the normal tissues but evidently abundant in the fibrous connective tissue of inflammatory and neoplastic lesions. In the present study, the close relationship between the distribution of FXIIIa-containing cells and that of collagenous components is demonstrated. The role that FXIIIa containing cells play in the process of fibrosis is discussed. PMID- 2570391 TI - T-lymphocyte subpopulations in acute unilateral optic neuritis. AB - The authors prospectively analyzed T-lymphocyte subpopulations in the peripheral blood of nine patients with acute unilateral optic neuritis and compared them with 25 controls without neurologic disease. The presence or absence of alterations in circulating T-cell subsets has not been examined previously in patients with isolated optic neuritis. The authors found the mean ratio of inducer (CD4) to suppressor (CD8) T-lymphocytes was 2.07 +/- 0.51 for the group with optic neuritis, statistically indistinguishable from a value of 1.78 +/- 1.04 for the control group. Multiple sclerosis (MS) subsequently developed in one patient. Her inducer/suppressor T-cell ratio was initially 2.66, but progressively increased to 3.68 concomitant with the clinical manifestation of focal neurologic signs. Although optic neuritis may be the initial clinical sign of MS, the periodic alteration of circulating T-lymphocytes increasing the inducer/suppressor T-cell ratio in MS was not observed in those with isolated optic neuritis. PMID- 2570393 TI - [The effect of mineral fertilizers on mosquito larvae (Culicidae)]. AB - The effect of superphosphate, nitrophoska and ammophos on larvae of Culicidae was studied under laboratory and field conditions of Novgorod Province. Superphosphate better than other fertilizers acidifies water, while nitrophoska and ammophos increase the ammonia contents of 27 to 240 fold that affects lethally the larvae. 0.4-0.5% and 1% concentrations of mineral fertilizers are a lethal dose for I-II instar and III-IV instar larvae, respectively. PMID- 2570394 TI - Major sperm protein and actin genes in free-living and parasitic nematodes. AB - The DNA from a number of free-living and parasitic nematode species was examined to determine the genomic number and distribution of DNA sequences encoding two evolutionarily conserved proteins; the major sperm protein (MSP) and nematode actin. Ascaris and Caenorhabditis MSP cDNA sequences and Ascaris genomic actin sequences were used to probe Southern blots of Eco RI and Hin d III digested nematode DNA. The number of MSP genes varied widely between the 1 MSP gene in Ascaris and the 60 MSP genes in Caenorhabditis. Filarial nematodes appeared to have 1-4 MSP genes while the plant and insect parasitic species showed from 5-12 MSP-hybridizing restriction fragments. Mammalian intestinal parasites showed between 1 and 13 bands hybridizing with the MSP probes. Blots probed to estimate the number of actin genes showed that, with the exception of Ascaris which contains more than 20 germ line sequences that encode actin, all of the nematodes tested had between 3 and 9 bands that hybridized to the Ascaris genomic actin probe. The possible use of highly conserved sequences such as MSP and actin to differentiate between nematode species in diagnostic and taxonomic studies is discussed. PMID- 2570395 TI - Kupalov's concept of shortened conditional reflexes: psychophysiological and psychopharmacological implications. AB - In the regulation of cortical excitability tonus, Kupalov described a particular acquired, learned mechanism, the shortened conditional Reflex (SCR). SCR is an essential mechanism by which an adequate cortical tonus is established by environmental cues as an anticipatory set-up preparing the individual qualitatively and quantitatively for expected forthcoming events. An original physiological example is given, the conditional "transfer" of motor behavior at cortical electrical stimulation. Three psychopharmacological implications are presented, namely environmental-dependent, stress-related events, environmental dependent drug conditioning, and drug-tolerance experiments. Future eventual psychophysiological and psychopharmacological lines of research related to the shortened conditional reflex concept are discussed. PMID- 2570396 TI - [Preoperative cytologic diagnosis of medullary carcinoma of the thyroid]. AB - The Authors emphasize the importance of preoperative aspiration cytology in the diagnosis of 5 medullary types out of 58 thyroid carcinomas observed in the Institute of Pathology of the Universita Cattolica del S. Cuore of Rome in the period 1983-88. All the cases were diagnosed preoperatively by mean of a fine needle aspiration cytological specimen; the case where morphological diagnosis was doubtful were studied with immunohistochemical techniques for calcitonin and CEA. Out of all cases, they report one case of multiple endocrine adenomatosis type II with medullary thyroid carcinoma and pheochromocytoma and another one with cervical node metastases without a clinically detectable thyroid swelling. Therefore, the authors propose the fine-needle aspiration cytology as a simple and important mean in the preoperative diagnostic procedure in case of medullary thyroid carcinoma. PMID- 2570397 TI - Developmental differences in neurohumoral modulation of the cat lower esophageal sphincter. AB - It is possible that age-related differences in gastroesophageal function during infancy may predispose the infant to abnormalities. We have therefore evaluated developmental differences in neurohumoral modulation of lower esophageal sphincter function in the cat. These studies were performed in an in vitro setting. In the adult cat, lower esophageal sphincter smooth muscle and postganglionic inhibitory neurons are more sensitive to muscarinic agonists than in the kitten. The circular smooth muscle of the adult lower esophageal sphincter is also more responsive to neurally mediated relaxation, brought about by electrical stimulation. However, there is no difference in response to exogenous vasoactive intestinal peptide between the two age groups. These data suggest that developmental differences in neurohumoral modulation of the lower esophageal sphincter occur in the cat. PMID- 2570398 TI - Heterogeneity of defects in mitochondrial acetoacetyl-CoA thiolase biosynthesis in fibroblasts from four patients with 3-ketothiolase deficiency. AB - Deficient mitochondrial acetoacetyl-CoA thiolase in fibroblasts from four patients with 3-ketothiolase deficiency was studied using immunochemical methods. We also examined fibroblasts from two heterozygotes, the mother and the brother of the case 1 patient, identified on the basis of the results of the enzyme activity measurements, using 2-methylacetoacetyl-CoA as substrate. The results were as follows: 1) in fibroblasts from all four patients, the thiolase activity using acetoacetyl-CoA was not activated by K+, although that of the controls and the heterozygotes was activated about 2-fold. 2) by immunoblot analyses, mitochondrial acetoacetyl-CoA thiolase was not detectable in fibroblasts from cases 2 and 3, although a very faint band was seen in tissues from cases 1 and 4. However, the band of mitochondrial 3-ketoacyl-CoA thiolase was clearly detected in all patients to the same extent as in the controls. 3) mitochondrial acetoacetyl-CoA thiolase was observed after pulse labeling for 1-h and a 72-h chase period in three cell lines (cases 1, 2, and 4), but was fainter compared to the controls. In another cell line (case 3), a fluorographic band at the same position was detected following a 1-h pulse, but disappeared following a 6-h chase. These results demonstrate heterogeneity in the enzyme defect resulting in a deficiency of mitochondrial acetoacetyl-CoA thiolase in fibroblasts from patients with 3-ketothiolase deficiency. PMID- 2570399 TI - Detectability of thyroid anti-microsomal antibodies, changes in thyroid stimulating immunoglobulins (TSI) and thyrotropin-binding-inhibiting immunoglobulins (TBII) during methimazole treatment of Graves' disease patients. AB - The prognostic value of determination of different antibodies in Graves' disease patients is questionable. The authors simultaneously assessed the generation of cAMP, the TSH-receptor binding inhibitory assay and the detectability of anti microsomal antibodies by indirect immuno-fluorescence. The tests were performed before, during and after methimazole treatment. During a 12 months' medication all 22 patients became euthyroid. Six months after withdrawal of the drug, 15 patients were still euthyroid (Group A); 7 relapsed (Group B). Patients showing enhanced activities by all three methods, relapsed (5 out of 7 cases of Group B). The results indicate that simultaneous determination of TSI, TBII and anti microsomal antibodies are of high prognostic value for relapses. These data should be taken into consideration for the further therapy. PMID- 2570400 TI - Treatment of multiple myeloma by high dose chemotherapy, total body irradiation and autologous blood stem cell autograft. AB - Fourteen patients with stage III aggressive multiple myeloma were treated by high dose chemotherapy (carmustine, etoposide and melphalan) and total body irradiation. This procedure was followed by autografting using blood stem cells previously collected by leukapheresis performed during recovery of cytotoxic drug induced bone marrow aplasia. One patient died from cerebral bleeding at day 40. All other patients, including 9 whose disease was refractory to conventional treatments, achieved an impressive tumor mass reduction (over 90% in 12 cases). Two patients relapsed and died 12 and 15 months after the graft; another one relapsed but is still alive at 24 months. Two to 23 months (median 12 months) after the autograft, 10 patients are well either in apparent complete remission (2 cases) or with a state of stable minimal residual disease. Autologous blood derived hematopoietic stem cells induced successful and sustained engraftment in the 13 evaluable patients. The cells collected by leukapheresis were studied for clonal Ig genes rearrangements indicative of circulating tumoral cells (or clonal B precursors) and none were found. Although preliminary, these results appear promising and high dose chemotherapy followed by autologous blood stem cell graft deserves consideration in young patients with multiple myeloma. PMID- 2570401 TI - Detection of novel genetic markers by mismatch analysis. AB - Chemical mismatch detection has been used to identify previously unknown genomic sequence variations that represent a new source of markers for genetic analysis. The approach detects all types of sequence changes, and therefore overcomes the limitation of restriction analysis, which identifies only a small fraction of the available sequence variations. Three new markers identified at the 3' end of the human dystrophin gene result from variable numbers of exact tandem repeats of 4bp (two examples) or 5bp (one example). None of these would have been detected as restriction fragment length polymorphisms by established procedures. PMID- 2570403 TI - The 5S rRNA-histone repeat in the crustacean Artemia: structure, polymorphism and variation of the 5S rRNA segment in different populations. AB - 5S rRNA genes are linked to the histone genes in the 13 populations of the crustacean Artemia that we have studied. In all cases, two types of repeat units are found. Southern blot analysis of all populations shows that they can be grouped into three classes: a) American bisexuals; b) Eurasian bisexuals, and c) parthenogenetic organisms (all from Eurasia). Restriction analysis of a bisexual population from San Francisco Bay shows that the two repeat units are of 9.0 and 8.5 kb (with minor heterogeneities of restriction sites). In parthenogenetic organisms, the two repeat units are of approximately 12 kb. Sequencing data from the region of the 5S rRNA from the San Francisco Bay population, shows that in both types of units, the single 5S rRNA gene (315 bp in length), is located 430 bp downstream the 3' regulatory sequences of the H2A gene, the last gene in the histone cluster. We have isolated three clones that contain 5S rRNA sequences. Two of them (one from an American bisexual and the other from a parthenogenetic population) contain histone and 5S rRNA genes, both with the same transcriptional polarity. The third clone, lacking histone genes, is likely to be an orphon derived from the parthenogenetic population. PMID- 2570404 TI - KpnI RFLP for the human CSF-1 gene. PMID- 2570405 TI - A BstXI polymorphism detected by the factor VIII genomic probe p.482.6 (F8C). PMID- 2570406 TI - Two linked RFLPs at chromosomal band 1p32. PMID- 2570402 TI - Variation in G + C-content and codon choice: differences among synonymous codon groups in vertebrate genes. AB - The relationship between G + C-content and codon usage in genes of human, mus, rat, bovine and chicken nuclear genomes was investigated. Correlation and lineal regression analyses were carried out on plots that related the frequency of each codon within each synonymous codon group to the G + C-content of the coding sequence as a whole. Under GC pressure, in most of the quartet codon groups there is a preferential choice of the C-ending codon, except in leucine and valine codon groups where the choice of the G-ending codon is preferred. Among ducts, the choice of codons specifying phenylalanine and glutamate shows the strongest dependence on G + C-content. The relationship found between G + C-content and codon usage in these genomes correlate with taxonomic distance. PMID- 2570407 TI - Three different insertion polymorphisms 3' to the human apolipoprotein A-IV gene. PMID- 2570408 TI - A PvuII polymorphism near the 5' end of the type II procollagen gene [COL2A1]. PMID- 2570409 TI - A TaqI RFLP detected by the probe VK45C6 [D16S131] at 16p13.11. PMID- 2570410 TI - An SstI polymorphism for the human muscarinic acetylcholine receptor gene, m4 (CHRM 4). PMID- 2570411 TI - A frequent DHFR polymorphism detected by an intron fragment. PMID- 2570412 TI - HincII RFLP in the human gene for the heavy neurofilament subunit (NF-H). PMID- 2570413 TI - An EcoRI polymorphism of the human von Willebrand factor cDNA (VWF). PMID- 2570414 TI - Common injuries of the foot. Often more than 'just a sprain'. AB - While most foot injuries heal without treatment, failure to recognize and treat some can have disastrous consequences. The exact mechanism of injury must be determined for accurate diagnosis. Many injuries, such as plantar fasciitis, "pump bump," sesamoiditis, and stress fractures, are the result of cumulative, repetitive stress rather than of an acute event. Others, such as injuries to tendons, may be chronic or acute. The foot is susceptible to numerous types of acute trauma, including sprains, fractures, dislocations, crushing, freezing, thermal injury, puncture wounds, and penetration by foreign bodies. Special care is required to minimize the danger of serious complications when treating foot injuries in diabetic patients. PMID- 2570415 TI - Adipose tissue deposition and cellularity in cimaterol-treated female broilers. AB - The effect of feeding a beta agonist, cimaterol, on carcass yield and composition and adipose tissue development was studied in female broilers. Cimaterol was provided at .25 ppm in the feed starting at 4 wk of age. Birds were sacrificed at 4, 6, 8, 10, 12, and 16 wk for determination of eviscerated carcass yield, carcass composition, and growth and cellularity of the abdominal, neck, back, and sartorial adipose depots. Carcass yield was significantly (P less than .01) greater in treated birds than in controls (70.0 vs. 72.8%) at 10 wk of age. Moisture percentage and protein decreased (P less than .05) less rapidly in treated carcasses than in control carcasses with increasing age. Lipid percentage increased (P less than .05) in treated carcasses less rapidly than in control carcasses. The weight of the abdominal fat pad was significantly (P less than .05) lower in treated birds than controls at 16 wk. There were no significant treatment effects on live weights, skeletal growth, or liver weights at any age. The composition of the pectoralis minor was not affected by the treatment. Frequency distributions of adipocyte diameters from the neck and thigh differed from those of the abdominal and back depots at 16 wk. Adipocyte diameter appeared to be more sensitive to the dietary treatment than adipocyte numbers per depot. Both adipocyte size and numbers in each depot continued to increase with increasing age of the broilers. The abdominal fat pad showed the largest percentage increase in cell numbers from 4 to 16 wk of age. PMID- 2570416 TI - New positive inotropic agents acting by phosphodiesterase inhibition or alpha 1 adrenergic stimulation. AB - New knowledge and insight into cardiac muscle physiology and pharmacology together with the development of novel drugs may change the treatment of congestive heart failure in the future. This article reviews two newer inotropic mechanisms currently under investigation. It will place special emphasis on the mechanisms of action of selective cardiac phosphodiesterase inhibition and of alpha 1-adrenoceptor mediated stimulation of the phosphoinositide pathway. PMID- 2570417 TI - Control of gastric acid secretion by histamine H2 receptor antagonists and anticholinergics. AB - The control of gastric secretion may be obtained by means of several pharmacological compounds: histamine H2 receptor antagonists and anticholinergics are so far the most widely employed drugs in pharmacological experiments and in clinical practice. The H2 blockers are able to inhibit the secretory response to histamine, acetylcholine and gastrin; they are effective and safe and at present they have been employed in several million patients suffering from diseases characterized by acid hyperproduction. The compounds available on the market are cimetidine, ranitidine, famotidine and nizatidine; however, approximately 11,000 compounds of the family have been synthesized and about 20 of these are under clinical evaluation. The blockade of H2 receptors is the primary action of these drugs; however, they possess also secondary actions which may represent untoward effects but in some cases may be actually useful (increase in prostaglandin synthesis, inhibition of LTB4 synthesis, etc.) The 'classic' anticholinergics appear to be decidedly less important mainly from a therapeutic point of view. However, the new compounds like pirenzepine and telenzepine, which block specifically the so-called M1 receptors located in the ganglia of the myenteric plexus, may represent an alternative to the H2 blockers since they are virtually devoid of the untoward reactions typical for the old atropine-like compounds (dry mouth, mydriasis, tachycardia, etc.) Both H2 blockers and the new antimuscarinic compounds may have effects not only on the parietal cells but also on other sites (G-cells, histaminocytes, D-cells) which control the function of the parietal cells themselves. PMID- 2570418 TI - Effects of amperozide in two animal models of anxiety. AB - The novel psychotropic agent amperozide (amp) was investigated in two different rat anxiety models, Vogel's conflict test (VT) and Montgomery's conflict test (MT). In the VT, amp in lower doses (0.2-0.6 mg/kg subcutaneously) increased the number of shocks accepted, as compared to controls. Pretreatment with the specific benzodiazepine receptor antagonist Ro 15-1788 (10.0 mg/kg orally) or the GABA-A receptor antagonist bicuculline (2.0 mg/kg intraperitoneally) antagonized the anticonflict effect of amp (0.4 mg/kg subcutaneously). At the highest dose tried (2.0 mg/kg subcutaneously) amp instead decreased the number of shocks accepted. Both 0.4 mg/kg and 2.0 mg/kg of amp raised the shock threshold, as compared to controls. The latter dose also decreased the motivation to drink. Pretreatment with Ro 15-1788 (10.0 mg/kg orally) did not significantly change the shock threshold or the drinking motivation, as compared to the group receiving amp alone. In the MT, amp (0.05-0.1 mg/kg subcutaneously) increased the percentage time spent in the open arms, while no changes were seen in the number of entries made into these arms. After higher doses (0.4-0.8 mg/kg subcutaneously) no differences, as compared to controls, were observed. Amp (1 nM 10microM) exhibited no affinity for 3H-flunitrazepam binding sites in mouse forebrain membranes in vitro. Taken together, the present data suggest that amp in low doses produces anticonflict (anxiolytic-like) effects. These effects appear to be mediated through an indirect (via 5-HT and/or DA systems?) activation of the GABA/benzodiazepine chloride ionophore receptor complex. PMID- 2570419 TI - [Continuous subcutaneous infusion of beta 2 receptor agonist in asthma in infants]. PMID- 2570420 TI - Mammalian multidrug-resistance gene: correlation of exon organization with structural domains and duplication of an ancestral gene. AB - Analysis of the nucleotide and deduced amino acid sequences of the biologically active mouse mdr1 cDNA clone indicates that the protein is formed by two highly homologous halves, each containing six putative transmembrane domains and a nucleotide-binding site. The duplicated unit shows high sequence homology to the proposed energy-coupling subunit of bacterial periplasmic transport proteins. We have cloned and characterized the mouse mdr1 gene and have analyzed the genomic organization of the two homologous halves forming the mdr1 protein. The gene spans 68 kilobases, is split into 28 exons, and the two homologous halves are encoded by 14 and 13 exons. The transcriptional initiation site of the gene has been mapped and putative TATA and consensus CAAT sequences have been found at positions -27 and -83, respectively. Discrete structural domains of the mdr1 protein are encoded by separate exons: Ten of the 12 putative transmembrane domains are encoded by individual exons and the two nucleotide-binding sites are each encoded by three exons. The exon/intron organization of the gene is conserved in the two highly homologous regions encoding the nucleotide-binding sites. The conservation of certain pairs of introns, together with the high degree of sequence homology, indicate that the mouse mdr1 gene originated from the duplication of an intron-containing ancestral gene. PMID- 2570421 TI - Pathway of proton transfer in bacterial reaction centers: replacement of glutamic acid 212 in the L subunit by glutamine inhibits quinone (secondary acceptor) turnover. AB - The mechanism of proton transfer in the reaction centers (RCs) from Rhodobacter sphaeroides was investigated by site-directed mutagenesis. Replacement of Glu-212 of the L subunit, a protonatable residue located near the secondary acceptor (QB) binding site, by glutamine reduced the in vitro electron turnover from cytochrome c to 2,3-dimethoxy-5-methylbenzoquinone (UQ0) by a factor of 25. The electron transfer rate to QB remained essentially unimpaired. Consequently, it is postulated that the reduced turnover in the mutant is due to a reduced rate of proton transfer to QB2-. The lack of pH dependence of the forward electron transfer rate DQA-QB----DQAQB- and the back reaction rate D+QAQB- ----DQAQB (where D = primary donor and QA = primary acceptor) in the mutant RC indicate that the observed pH dependence in the native RC is due to Glu-212, which has an anomalously high pKa value of 9.5 +/- 0.3. These results support the involvement of Glu-212 as a proton donor to reduced QB. PMID- 2570423 TI - Propulsion of a fin whale (Balaenoptera physalus): why the fin whale is a fast swimmer. AB - Measurements of an immature fin whale (Balaenoptera physalus), which died as a result of entrapment in fishing gear near Frenchmans Cove, Newfoundland (47 degrees 9' N, 55 degrees 25' W), were made to obtain estimates of volume and surface area of the animal. Detailed measurements of the flukes, both planform and sections, were also obtained. A strip theory was developed to calculate the hydrodynamic performance of the whale's flukes as an oscillating propeller. This method is based on linear, two-dimensional, small-amplitude, unsteady hydrofoil theory with correction factors used to account for the effects of finite span and finite amplitude motion. These correction factors were developed from theoretical results of large-amplitude heaving motion and unsteady lifting-surface theory. A model that makes an estimate of the effects of viscous flow on propeller performance was superimposed on the potential-flow results. This model estimates the drag of the hydrofoil sections by assuming that the drag is similar to that of a hydrofoil section in steady flow. The performance characteristics of the flukes of the fin whale were estimated by using this method. The effects of the different correction factors, and of the frictional drag of the fluke sections, are emphasized. Frictional effects in particular were found to reduce the hydrodynamic efficiency of the flukes significantly. The results are discussed and compared with the known characteristics of fin-whale swimming. PMID- 2570422 TI - Immunogenicity and efficacy testing in chimpanzees of an oral hepatitis B vaccine based on live recombinant adenovirus. AB - As a major cause of acute and chronic liver disease as well as hepatocellular carcinoma, hepatitis B virus (HBV) continues to pose significant health problems world-wide. Recombinant hepatitis B vaccines based on adenovirus vectors have been developed to address global needs for effective control of hepatitis B infection. Although considerable progress has been made in the construction of recombinant adenoviruses that express large amounts of HBV gene products, preclinical immunogenicity and efficacy testing of candidate vaccines has remained difficult due to the lack of a suitable animal model. We demonstrate here that chimpanzees are susceptible to enteric infection by human adenoviruses type 7 (Ad7) and type 4 (Ad4) following oral administration of live virus. Moreover, after sequential oral immunization with Ad7- and Ad4-vectored vaccines containing the hepatitis B surface antigen (HBsAg) gene, significant antibody responses to HBsAg (anti-HBs) were induced in two chimpanzees. After challenge with heterologous HBV, one chimpanzee was protected from acute hepatitis and the other chimpanzee experienced modified HBV-induced disease. These data demonstrate the feasibility of using orally administered recombinant adenoviruses as a general approach to vaccination. PMID- 2570425 TI - Analysis of non-disjunction in human trisomic spontaneous abortions. PMID- 2570424 TI - Studying nondisjunction of chromosome 21 with cytogenetic markers on the short arm and DNA markers encompassing the long arm. PMID- 2570426 TI - The beta-receptor, atheroma and cardiovascular damage. AB - In man a close interrelationship exists between hyperadrenergic states, myocardial ischemia, necrosis, infarction and sudden cardiac death. Persistent high catecholamine levels may also be associated with increased vascular endothelial turnover and permeability to calcium and lipoproteins, increased blood velocity, abnormal blood flow patterns and atheroma formation. There are thus good reasons to predict a cardiovascular protective effect of beta-blockers. Animal data indicate that in spite of apparently adverse plasma lipoprotein changes beta-blockers retard atheromatous plaque formation under conditions of high cholesterol diet with or without stress. A slow heart rate, as well as a reduction in calcium influx and inhibition of both esterification of arterial wall cholesterol (by ACAT) and endothelial permeability to lipoproteins, may be central to this process. Beta-blockers benefit a spectrum of conditions related to the atheromatous process and myocardial necrosis. These are silent ischemia; stable (including mixed), unstable and preinfarction angina; periinfarction events (including myocardial rupture and dissection of the ascending aorta); and myocardial necrosis associated with stress conditions such as head injuries and subarachnoid hemorrhage. In one study coronary deaths in hypertensive men, particularly in smokers, were significantly reduced by metoprolol (a beta 1 selective blocker) compared to a diuretic. In contrast in the MRC study of mild hypertension only nonsmoking men with mild to moderate hypertension who received a nonselective beta-blocker appeared to experience fewer myocardial infarctions. Recent clinical data showed that moderate-severe hypertensives who were optimally controlled by atenolol-based treatment over a 10-year period were less likely to die from myocardial infarction than those suboptimally controlled, irrespective of a rise in serum triglyceride levels. Thus the net effect of acute beta blockade in hyperadrenergic states, including myocardial infarction, is to limit cardiovascular damage. Chronic beta-blockade inhibits atheroma formation (in animals) and beneficially modifies the incidence of stroke and myocardial infarction, which in man are the long-term consequences of hypertension. PMID- 2570427 TI - Sulphasalazine and related drugs in rheumatoid arthritis. PMID- 2570428 TI - Spermatozoa of the deep-sea cephalopod Vampyroteuthis infernalis Chun: ultrastructure and possible phylogenetic significance. AB - Sperm ultrastructure in the rare deep-sea cephalopod Vampyroteuthis infernalis is described, based on formalin-fixed material held in the Australian Museum (Sydney). The species is the sole member of the coleoidean order Vampyromorpha, which represents a level of organization intermediate between that of the Sepioidea-Teuthoidea and the Octopoda. Spermatozoa of Vampyroteuthis, the simplest observed in any cephalopod, exhibit the following features: (1) a spheroidal acrosome lacking any complex substructure; (2) a short (8.5 microns) fusiform nucleus with a deep (2.2-2.5 microns) basal invagination (containing an extensive plug of dense material); (3) two triplet centrioles arranged parallel to the sperm longitudinal axis; (4) a short (1 micron) midpiece composed of a triangular cluster of mitochondria surrounding the centrioles; and (5) a tail (length 130-135 microns) that is continuous with one of the centrioles (here considered as a 'distal' centriole). An annulus and membranous skirt are absent, though the coarse fibres do fuse into a ring at the tail-midpiece junction). These cells show some resemblance to sperm or spermatids of sepioids and teuthoids (spheroidal acrosome, short nucleus) but are also remarkably similar to mid-spermatids of Octopus (with the exception of the uncondensed nucleus in Octopus spermatids). Sperm morphology supports the current assignment of Vampyroteuthis to a separate coleoidean order--Vampyromorpha--and also suggests that a close link exists between the Vampyromorpha and Octopoda. PMID- 2570429 TI - RBC and plasma choline in neuroleptic-treated schizophrenic patients. AB - We measured red blood cell (RBC) choline and plasma choline concentrations in 27 chronic schizophrenic inpatients and 23 normal controls. Both blood choline measures had a significant test-retest reliability in patients whose neuroleptic status remained unchanged over 1 month. RBC choline concentration was significantly lower in patients medicated with neuroleptics and cogentin. Patients with a low RBC choline and a low RBC/plasma choline ratio were on significantly higher doses of medication and had higher scores on the hostility/suspiciousness subscale of the Brief Psychiatric Rating Scale. RBC choline increased when neuroleptics were discontinued. Blood choline measures were also compared among medication-free schizophrenic patients, inpatients with other diagnoses, and normal controls. No significant differences were seen among these groups for any choline measure, although the schizophrenic patients showed greater variability. Medication-free schizophrenic patients with such clinical factors as tardive dyskinesia and abnormalities on computed tomography contributed to this variability. Age was positively correlated with plasma choline. PMID- 2570431 TI - Blockade of acquisition of one-way conditioned avoidance responding by haloperidol and metoclopramide but not by thioridazine or clozapine: implications for screening new antipsychotic drugs. AB - The classical neuroleptic drugs haloperidol and pimozide have a strong disruptive effect on the acquisition of conditioned avoidance responding (CAR), yet have relatively little impact on the performance of previously acquired responses. Separate experiments compared the effects of haloperidol, two atypical neuroleptics, thioridazine and clozapine, and a substituted benzamide, metoclopramide, on one-way avoidance by rats. Thioridazine (10-50 mg/kg) and clozapine (1.25-10.0 mg/kg) disrupted both acquisition and performance of CAR. In contrast, haloperidol (0.075-0.150 mg/kg) and metoclopramide (5.0-7.5 mg/kg) completely blocked the acquisition of CAR, yet initially produced only a slight disruption in the performance of a previously acquired response. The ineffectiveness of the atypical neuroleptics in producing a complete disruption of acquisition of CAR may be due to the anticholinergic properties of these drugs. Alternatively, the differences between metoclopramide and the atypical neuroleptics may be due to a preferential effect of metoclopramide on striatal or amygdaloid dopamine neurotransmission. These results suggest that caution should be exercised in using CAR as an animal model for assessing the antipsychotic potential of new pharmacological agents. PMID- 2570430 TI - Pharmacokinetics of long-acting injectable neuroleptic drugs: clinical implications. AB - The authors review the literature regarding the pharmacokinetics of long-acting injectable neuroleptic drugs (LINS). There are important differences between LINS and oral neuroleptics that affect their pharmacokinetics. By avoiding first pass metabolism in gut and liver, LINS result in lower circulating concentrations of metabolites than are found after oral administration. In addition, LINS take more time to reach a stable steady state than their oral counterparts. The clinical significance of these pharmacokinetic properties is discussed. The authors recommend that when patients are being changed from oral neuroleptics to LINS, that this conversion be done gradually over several months. PMID- 2570432 TI - Differential tolerance to cataleptic effects of SCH 23390 and haloperidol after repeated administration. AB - The development of tolerance to the cataleptic effect of the selective D-1 antagonist SCH 23390 (0.5 mg/kg/day SC or 0.1 mg/kg/day SC) and haloperidol (1 mg/kg/day SC) during repeated administration was investigated. Catalepsy in rats was measured using the horizontal bar method. SCH 23390 induced a dose-related cataleptic effect of short duration, whereas the cataleptic effect of haloperidol appeared more slowly and lasted longer. Marked tolerance to the cataleptic effect of haloperidol developed already 6 days from the beginning of the treatment. The cataleptic effect of the higher dose regimen of SCH 23390 was also significantly reduced after 6 days' treatment. However, unlike haloperidol, this subacute tolerance was gradually reversed and was no longer significant after 12 and 18 days. The cataleptic response to the lower dose of SCH 23390 (0.1 mg/kg/day) was not significantly altered during the treatment and no initial catalepsy tolerance was observed with this dose regimen. These results suggest that different mechanisms are involved in the expression of cataleptic behaviour during chronic treatment with SCH 23390 and classical antipsychotics, such as haloperidol. PMID- 2570433 TI - Scopolamine and benzodiazepine models of dementia: cross-reversals by Ro 15-1788 and physostigmine. AB - The muscarinic antagonist scopolamine and the benzodiazepine lorazepam both produce transient impairments in memory and attention in normal volunteers. These impairments can be reversed by appropriate agents such as the cholinesterase inhibitor physostigmine in the case of scopolamine or the benzodiazepine antagonist Ro 15-1788 in the case of lorazepam. In this paper we investigated the pharmacological specificity of these reversals by examining the interactions of scopolamine and Ro 15-1788 and of lorazepam and physostigmine. There was no evidence that the effects of scopolamine and lorazepam on cognitive function could be attenuated by Ro 15-1788 and physostigmine, respectively. The results are discussed in terms of pharmacological models of Alzheimer's disease. PMID- 2570435 TI - Rats administered chronic neuroleptics develop oral movements which are similar in form to those in humans with tardive dyskinesia. AB - Oral movements (OMs) in rats chronically administered haloperidol (HAL), fluphenazine (FLU), or no drug were recorded using a computerized video analysis system which measured the distance between two fluorescent dots painted above and below the rat's mouth. The resulting data was analyzed using fast-fourier analysis. Following an initial period of sedation (decreased energy at all frequencies), the drugged animals (and especially the FLU animals) began to show increased oral movements of 1-2 Hz, an effect which increased substantially upon drug withdrawal. This is precisely the altered energy spectrum observed in humans with tardive dyskinesia. PMID- 2570436 TI - SKF 38393 potentiates yawning induced by LY 171555: further evidence against the autoreceptor hypothesis of yawning. AB - The effect of concurrent D-1 receptor stimulation by SKF 38393 on the expression of yawning elicited by D-2 receptor stimulation with LY 171555 was studied in the rat. A low dose of SKF 38393 (2.5 mg/kg SC), while failed to elicit yawning, potentiated the effectiveness of LY 171555 in eliciting yawning at all the doses tested (12.5, 25 and 50 micrograms/kg SC) and this effect was abolished by SCH 23390 (0.012 mg/kg SC). The results indicate that in analogy with typical post synaptic dopaminergic effects (hypermotility-stereotypy), yawning elicited by a D 2 agonist is facilitated by concurrent stimulation of D-1 receptors and therefore is consistent with previous evidence that yawning in response to a D-2 agonist is not mediated by autoreceptors. PMID- 2570434 TI - Centrally administered neuropeptide Y (NPY) produces anxiolytic-like effects in animal anxiety models. AB - Effects of intracerebroventricular (ICV), neuropeptide Y (NPY) (0.2-5.0 nmol) and its C-terminal 13-36 amino acid (AA) fragment (0.4-2.0 nmol) have been examined with respect to anxiolytic properties in two rat anxiety models, Montgomery's conflict test (MT), and Vogel's drinking conflict test (VT). In the MT, 1.0 and 5.0 nmol NPY abolished the normal preference for the closed arms of the maze. At 5.0 nmol, the total number of entries made into both closed and open arms was decreased by 50%. In the VT, both 0.2 and 1.0 nmol NPY markedly increased the number of shocks accepted. The effect of 5.0 nmol NPY was less pronounced. In control experiments, NPY (0.2 nmol) did not affect pain sensitivity or thirst. Pretreatment with the selective alpha 2-adrenergic receptor antagonist idazoxan, at a dose which by itself did not affect behaviour (2.0 mg/kg), antagonized the effect of 1.0 nmol NPY in the VT. NPY 13-36 was without significant effect in both models. The results suggest that NPY exerts anxiolytic-like effects, and that these effects are mediated through an interaction with noradrenergic systems. Higher doses of NPY produce sedation and ataxia, which decrease overall activity in the MT, and interfere with the ability fully to express behaviourally the anxiolytic-like effect in the VT. The findings are discussed in relation to the noradrenaline hypothesis of anxiety, and to observations indicating involvement of NPY in the pathophysiology of major depression. PMID- 2570437 TI - Treatment of panic disorder: nonbenzodiazepine anxiolytics, including buspirone. PMID- 2570439 TI - Diagnostic imaging of lower extremity trauma. AB - Traumatic injuries to the lower extremity are a common occurrence in today's society and may result in significant morbidity and mortality if not appropriately treated. Adequate radiographic evaluation is crucial to the assessment of these injuries. This article reviews the radiographic features and imaging approaches to the commonly encountered injuries of the lower extremity. PMID- 2570438 TI - Zotepine vs. haloperidol in paranoid schizophrenia: a double-blind trial. PMID- 2570440 TI - Imaging of musculoskeletal trauma in three dimensions. An integrated two dimensional/three-dimensional approach with computed tomography. AB - Transaxial computed tomographic (CT) data can be used to generate images rapidly in the coronal and sagittal planes (two-dimensional CT) and create simulated three-dimensional images. The techniques and advantages of an integrated two dimensional/three-dimensional CT approach to musculoskeletal trauma are illustrated and discussed. PMID- 2570441 TI - An enzyme histochemical analysis of the different steps in thioacetamide induced liver carcinogenesis in rats. PMID- 2570442 TI - Antibodies against neuroactive amino acids and neuropeptides. A new two-step procedure for the preparation of carrierprotein linked amino compounds. PMID- 2570443 TI - Antibodies against neuroactive amino acids and neuropeptides. Combination of acetylcholinesterase cytochemistry with immunocytochemical double staining techniques for the demonstration of GABA and glutamate like immunoreactivities in thalamus, hippocampus, cerebellar and cerebral cortex of the rat. PMID- 2570444 TI - An LTC4 binding site in gastric mucosa is shared with glutathione. AB - Recent results from our laboratory and others have suggested a possible physiological functional role(s) for leukotrienes in gastric mucosa. In the present study 3H-LTC4 binds to washed rabbit gastric mucosal membranes at 4 degrees C with a Kd of 5 nM and a Bmax of 31.3 pmol/mg protein. Leukotrienes D4, E4, B4, oxidized glutathione (GSSG), cysteine, and mercaptoethanol were unable to displace 3H-LTC4 at 1 microM concentrations, while GSH inhibited binding with a Ki of 47 nM. Differential centrifugation of the membrane preparation to remove mitochondria resulted in Ki values for LTC4 and GSH of 14 and 23 nM, respectively. The similar binding affinities and competitive receptor binding kinetics for GSH and LTC4, the low affinity for other leukotrienes, and a Ki of 7 microM for hematin, a substrate for glutathione S-transferase, suggest that 3H LTC4 binds to a GSH site which does not discriminate between LTC4 and GSH. Membranes fractionated to remove mitochondria were assayed for glutathione peroxidase, gamma-glutamyltranspeptidase, and glutathione S-transferase as possible binding sites for LTC4. We were unable to detect enzyme activity for any of the three enzymes. The binding of LTC4 in gastric mucosa differs from other tissues with respect to the high affinity for GSH, and thus becomes an appropriate tissue in which to investigate the relationships between LTC4 and GSH. PMID- 2570445 TI - [Type 2 multiple endocrine neoplasm]. PMID- 2570446 TI - [Prenatal diagnosis in a family carrying the fragile X syndrome using recombinant DNA technics]. AB - The prenatal diagnosis in a family carrying the Fragile Chromosome X Syndrome (sex linked mental retardation) has been performed using Recombinant DNA techniques. The use of these techniques together with cytogenetics and prenatal diagnosis using chorial biopsy, have enabled the diagnosis of the disease on a male foetus on the twelfth week of pregnancy. Similarly, we have been able to confirm the carrier state of the mother (proven) and the grand mother, and furthermore, the non affected condition of an older son to this mother. The results obtained reveal the importance of the new molecular techniques applied to the diagnosis of certain diseases with a genetic origin. PMID- 2570447 TI - [Changes of liver enzymes during severe infections]. AB - To establish the prevalence of liver enzyme alterations in the course of serious infections of diverse origin, 112 patients were studied retrospectively for the levels of GOT, GPT, LDH, GGT, alkaline phosphatase (FA) and bilirubin (BIL). Cases in which the values might be altered due to causes other than the infectious process were previously excluded. The prevalence of affectation of each of the parameters was the following: GGT, 39.1%; LDH, 33.3%; FAL, 30.1%; GOT, 29.5%; GPT, 24.8%; BIL, 18.3%. Seventy percent had at least one of these values changed. No significant differences were found in the incidence or intensity of the analytic alterations in function of the origin of the of infection, which suggests that these modification are nonspecific. Bilirubin levels were found to be significantly higher in the patients who died. On realizing the linear correlation analysis between the diverse parameters studied, a good correlation was found between them, suggesting a common pathogenetic mechanism. PMID- 2570448 TI - Acute peripheral catecholaminergic changes in rat after MPTP and MPP+ treatment. AB - The effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and its main metabolite 1-methyl-4-phenylpyridinium ion (MPP+) on the peripheral catecholaminergic system of the rat were investigated. MPTP and MPP+ injections (20 mg/kg i.p.) caused a marked acute depletion of heart noradrenaline, up to 75% twelve hours after the administration, and a decrease of adrenal gland adrenaline. The time-course of the effect of MPTP and MPP+ is reported, together with a decrease in the tyrosine hydroxylase activity after MPTP treatment, more evident in the adrenal glands. Pargyline (50 mg/kg i.p.) is not able to prevent such a neurotoxic peripheral effect. PMID- 2570449 TI - Effects of selective and non-selective beta 1-blockade on renin secretion in the isolated rat kidney. AB - The participation of beta 1-adrenoceptor on renin secretion was studied in perfused rat kidney. Administration of propranolol, inhibited the renin release mediated by isoproterenol. Likewise, metoprolol and practolol, showed a similar potency to propranolol in inhibiting isoproterenol-induced renin secretion. These results suggest that the for isoproterenol-induced renin release in rats is a beta 1-type adrenoceptor. PMID- 2570450 TI - Is a simultaneous beta-blocker therapy a risk factor for enalapril-induced cough? PMID- 2570451 TI - [Chemical and pharmacologic aspects of benzodiazepines]. AB - Benzodiazepines (BZ) discovered by chance by the American chemist Sternbach have been indispensable drugs not only in treatment of psychic disorders but also as antiepileptics and myorelaxants. Their heterocyclic structure is essential for the spectrum of activities. Pharmacokinetic studies have unveiled that many BZ are prodrugs of active metabolites. Nearly all BZ are lipophilic amides which are rapidly and efficiently resorbed after oral administration. Some BZ are subjected to a first-pass effect. Protein binding varies from 80-90%. Distribution is rapid but plasmatic levels are often not well correlated with biologic action. Hepatic metabolism is important whereby oxidative pathways prevail. BZ act through amplification of the effects of the neurotransmitter y-aminobutyric acid (GABA). The extent of action is thus limited within narrow margins. Subtypes of BZ receptors found experimentally might indicate that certain actions i.e. anxiolysis arise via subtype specific binding. The development of more selective drugs, free of side effects, could therefore become possible. Newly developed BZ antagonists counteract most BZ-effects efficiently. A practical classification of BZ uses plasmatic elimination-halflife (t 1/2): 1. Long-acting substances (t 1/2: up to 100 hrs): Diazepam, Nitrazepam, Flurazepam, Bromazepam (most with active metabolites) 2. Substances of intermediate action (t 1/2: up to 30 hrs): Oxazepam, Lorazepam, Flunitrazepam, Temazepam (few active metabolites) 3. Short acting substances (t 1/2 up to 8 hrs): Triazolam, Nidazolam (few active metabolites) PMID- 2570452 TI - [Benefits and risks in using benzodiazepines from the viewpoint of pharmaco epidemiology with reference to substance dependence]. AB - Anxiety and sleep disturbances are frequent symptoms in medical practice. They still constitute the main indications for prescription patterns. The evaluation of risks and benefits deals with many methodological problems concerning terminology of dependence and pharmaco-epidemiology finally findings in the literature are reviewed under the aspects of diminishing the risk of dependence. PMID- 2570453 TI - [Benzodiazepine abuse--findings from a German state mental hospital]. AB - During the past few years the problem of over-prescribing of benzodiazepines has been increasingly discussed. However, up to now exact data on the extent of an abuse of these substances are relatively scarce and controversial. An analysis done at a German Psychiatric State Hospital from 1974 to 1983 regarding the frequency of the diagnosis "Benzodiazepine-dependence" showed that in only 150 cases (0.5%) out of almost 33,000 admitted patients was the diagnosis benzodiazepine-abuse justified, with an increase since the year 1980. During a prospective study of the quarterly incidence in 1984, 18.5% of the patients admitted met the criterion of long-term use of benzodiazepines (continuous use for more than three months). Sociodemographic and clinical data of these benzodiazepine-long term users are presented. PMID- 2570454 TI - [Problems in long-term benzodiazepine treatment]. AB - The risk of drug dependency following long-term use of benzodiazepines is reviewed and complemented with own experiences. Withdrawal from high- and low dose dependency was followed by systematically recording symptoms. Insomnia, a cardial symptom, was recorded in the sleep-laboratory during withdrawal. Benzodiazepines suppress the deep phases of sleep almost completely. In the course of withdrawal induced rapidly by a 50% reduction of the administered dose at five day intervals leading to substantial withdrawal symptoms sleep-EEG's improved considerably. The patients felt substantially better after withdrawal. PMID- 2570455 TI - [Perspectives of the 3d Basel Psychiatry Rounds]. AB - This work discusses the papers of the third symposium on psychiatry held in Basle. Herzka interprets substance abuse from a historical and cultural point of view as an escape from external tensions into an internal fight with the drug. The increase of tolerance of the many controversies and discrepancies in our culture could help to prevent drug abuse. The doctor is also in a clash between different cultures and social systems, where most values have become relative. In this situation the doctor will be tempted to only take some of the aspects of the bio-psychosociological model of psychiatric illnesses into account and to become highly specialized as to evade the conflict himself. The development of new benzodiazepines is progressing and the isolation of partial effect components, such as pure antianxiety and antiaggressive effects, is an important goal which has been partly reached. Benzodiazepines today belong to the most important medication for insomnia, restlessness, tension, anxiety present in different functional disorders, which not only concern psychiatry but medicine in general. A lot of patients need longterm treatment with benzodiazepines. This fact is contrary to the common recommendation to avoid benzodiazepine dependence. The one year prevalence of dependence is estimated to be, according to Ladewig, 0.1% of the population, which is, with regard to frequency and toxicity, compared to other substances, socially and medically relatively low. The therapeutic benefit is worth the risk of dependence. The prophylaxis for the reduction of drug dependence in the population is effective.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570456 TI - Drug interactions with quinolones. AB - Numerous drug interactions with the new 4-quinolone antimicrobial agents have now been established. Many, but not all, quinolones are extensively metabolized and can have inhibitory effects on the liver cytochrome P450 enzyme system, leading to reduced metabolism and clearance of certain other drugs that are normally thus eliminated. Examples include the highly significant interaction between enoxacin and theophylline and the interaction between ciprofloxacin and theophylline, which may also be important clinically. The quinolone-caffeine interaction does not usually cause problems. Absorption of all quinolones from the stomach and small intestine is greatly reduced by antacids containing magnesium or aluminium salts, including sucralfate, probably as a result of the formation of nonabsorbable chelates. Cimetidine can reduce the clearance of pefloxacin (but not of ciprofloxacin) through its effects on liver metabolism, although newer H2 inhibitors appear not to have these effects. Probenecid reduces the renal elimination of some quinolones by inhibiting tubular secretion. New evidence is now coming to light of interactions between certain nonsteroid antiinflammatory drugs (e.g., fenbufen), quinolones, and gamma-aminobutyric acid (GABA) receptors, producing increased cerebral excitation and, sometimes, epileptiform convulsions. PMID- 2570457 TI - ["We broke new ground". Interview by K. Venner]. PMID- 2570458 TI - [Research on neurons of the somatotropin release inhibiting hormone in th hypothalamus]. PMID- 2570459 TI - [Molecular biology research on ApoA-I]. PMID- 2570460 TI - Identification of the cystic fibrosis gene: genetic analysis. AB - Approximately 70 percent of the mutations in cystic fibrosis patients correspond to a specific deletion of three base pairs, which results in the loss of a phenylalanine residue at amino acid position 508 of the putative product of the cystic fibrosis gene. Extended haplotype data based on DNA markers closely linked to the putative disease gene locus suggest that the remainder of the cystic fibrosis mutant gene pool consists of multiple, different mutations. A small set of these latter mutant alleles (about 8 percent) may confer residual pancreatic exocrine function in a subgroup of patients who are pancreatic sufficient. The ability to detect mutations in the cystic fibrosis gene at the DNA level has important implications for genetic diagnosis. PMID- 2570461 TI - Molecular characterization of the human beta 3-adrenergic receptor. AB - Since the classification of beta-adrenergic receptors (beta-ARs) into beta 1 and beta 2 subtypes, additional beta-ARs have been implicated in the control of various metabolic processes by catecholamines. A human gene has been isolated that encodes a third beta-AR, here referred to as the "beta 3-adrenergic receptor." Exposure of eukaryotic cells transfected with this gene to adrenaline or noradrenaline promotes the accumulation of adenosine 3',5'-monophosphate; only 2 of 11 classical beta-AR blockers efficiently inhibited this effect, whereas two others behaved as beta 3-AR agonists. The potency order of beta-AR agonists for the beta 3-AR correlates with their rank order for stimulating various metabolic processes in tissues where atypical adrenergic sites are thought to exist. In particular, novel beta-AR agonists having high thermogenic, antiobesity, and antidiabetic activities in animal models are among the most potent stimulators of the beta 3-AR. PMID- 2570462 TI - Free rectus abdominis muscle flap: advantages in lower extremity reconstruction. AB - Major soft tissue losses of the distal portion of the leg usually require free tissue transfer for reconstruction. We have recently treated seven patients with free rectus abdominis muscle flaps. The rectus abdominis has several advantages over other muscles; these advantages simplify such reconstructions. Elevation of this muscle can be done simultaneously with recipient vessel dissection without the need for patient repositioning. The deep inferior epigastric vessels provide a reliable pedicle of one large artery and two veins of sufficient length to accomplish microvascular anastomosis outside the zone of injury without the use of vein grafts. The size and shape of the muscle conforms well to the defects of many wounds in the leg. There is no functional deficiency from the use of one muscle, and the donor site scar is relatively inconspicuous since the rectus abdominis is harvested through a midline abdominal incision. There have been two complications in this series, one intraoperative flap failure, and one case of recurrent osteomyelitis. There has been no donor site morbidity. We recommend the free rectus abdominis muscle flap for the reconstruction of many complex wounds of the distal portion of the leg. PMID- 2570463 TI - [A case with polyarteritis nodosa associated with acute renal failure and pericardial cyst]. AB - Polyarteritis nodosa (PN) is a disease characterized by vasculitis of multiple organs, and it is complications of kidney and heart are fatalistic factor. Myocardial infarction and pericarditis are well known as cardiac involvement of PN. The pericardial cysts are uncommon disease and it originated in the mesenchymal tissue during fetal life. There has been no report that pericardial cyst undergoing with PN. The present paper was the first report about case with enlargement of pericardial cyst caused by vasculitis. A 64-years old female was referred for acute renal failure and mediastinal tumor. The diagnosis of PN was confirmed with the renal biopsy showing the typically histological features of necrotizing angitis, renal angiography showing vascular irregularity in the renal arterioles and clinical features such as fever, hypertension and skin lesion. The mediastinal tumor was identified with enlargement of pericardial cyst diagnosed by computer tomography (CT) and magnetic resonance imaging (MRI), and ultrasonic cardiography (UCG) revealed pericardial effusion. Enlargement of pericardial cyst, pericardial effusion and acute renal failure were due to vasculitis as a partial manifestation of PN. Under the steroid administration, sequentially recovery of renal function and disappearance of pericardial cyst and pericardial effusion were observed. PMID- 2570464 TI - Gonadotrophin stimulation in children with abnormal sexual development. AB - Plasma levels of testosterone were measured by radioimmunoassay before and after 3 days' administration of human chorionic gonadotrophin (HCG) 2,000 IU/d in 34 prepubertal boys (6 normal, 3 with anorchia, 5 with micropenis, 8 with cryptorchidism, 6 with hypogonadotrophic hypogonadism and 6 with hypospadias). The increase in plasma testosterone value ranged from 2.4 nmol/l to 10.5 nmol/l in normal boys. There was negligible (P greater than 0.05) response in boys with anorchia compared with normal children. Subjects with micropenis showed subnormal response (P greater than 0.05). Children with cryptorchidism showed a normal increase in testosterone levels (P greater than 0.05), but in those with hypogonadotrophic hypogonadism and hypospadias the increase was variable (1.2 - 8.8 nmol/l) (P greater than 0.05). Although the HCG stimulation test is not diagnostic in the majority of the sexual disorders, it appears to be sensitive for diagnosing anorchia. PMID- 2570465 TI - Subdural empyema--an unusual presentation. A report on 2 cases. AB - Two cases of subdural empyema with unusual clinical features are described. Both had an undetermined primary source of infection, the onset was insidious and the course was rather indolent, and both cases occurred after steroid therapy. Subdural empyema is a potentially lethal condition if inappropriately managed. A high index of suspicion and prompt definitive treatment remain the cornerstones of successful management. The aetiology and pathogenesis of subdural empyema and the merits and demerits of various investigative procedures are discussed; the factors that influence prognosis are highlighted. PMID- 2570466 TI - Fade of the response to prolonged glutamate application in the rat hippocampal slice. AB - The effect of prolonged glutamate (GLU) application was examined on 60 CA1 pyramidal neurons in the in vitro rat hippocampal slice preparation. Continuous application of L-GLU, either by bath perfusion (0.5-2 mM) of the slices or iontophoresis (200 mM) into the dendritic region of the neurons, elicited a transient depolarization which faded to a mean of 53% of the initial peak amplitude despite continued exposure to the agonist. Membrane depolarization to aspartate (ASP) and the d-isomer of GLU also faded with time. In contrast, the depolarizing response to the excitatory amino acid agonists N-methyl-D,L aspartate (NMA), quisqualate (QUIS), and kainate (KA) did not fade significantly during continuous application. The fade of the GLU depolarization was not affected by the NMDA antagonist D-2-amino-5-phosphonovalerate (APV) or by blocking synaptic transmission with tetrodotoxin. At the time of maximum fade of the GLU depolarization, there was no change in input resistance or GLU reversal potential. In addition, fade of the response was not a consequence of changes in extracellular potassium concentration, GLU uptake mechanisms, or the electrogenic pump. The most likely explanation for fade is postsynaptic receptor desensitization. PMID- 2570467 TI - Human ventral mesencephalic xenografts to the catecholamine-depleted striata of athymic rats: ultrastructure and immunocytochemistry. AB - On the basis of animal studies, grafts of fetal human dopaminergic cells have been suggested as a therapy for Parkinson's disease. The purpose of this study was to characterize the ultrastructure and immunocytochemistry of human ventral mesencephalic xenografts placed into the catecholamine-depleted striata of athymic "nude" rats. Human fetal tissue was obtained from tissue fragments derived from elective abortions during the first trimester of pregnancy. Small pieces of the basal mesencephalon were grafted into the catecholamine-depleted striata of four athymic nude rats. The rats were allowed to survive from 3 to 6 months after grafting; following fixation, the striatal tissue containing the grafts was labeled with antibodies against tyrosine hydroxylase and serotonin. Immunocytochemistry revealed tyrosine-hydroxylase-like-immunoreactive (THLI) and serotoninlike-immunoreactive (5HTLI) cell bodies within the human grafts. Both 5HTLI and THLI fibers crossed the graft-host interface and innervated the previously lesioned striatum. Both types of fibers also entered the host cortex from the adjacent human graft. At the ultrastructural level, THLI and 5HTLI fibers and synaptic terminals were observed in the host neuropil. THLI and 5HTLI dendrites and axon terminals were also observed in the neuropil of the grafts themselves. THLI axon terminals are not normally present in the substantia nigra. The results of our study indicate that human xenografts can survive in the neuropil of the host striatum and form morphologically appropriate synapses within the host brain. PMID- 2570468 TI - Mutagenic lipid peroxides from edible oils. AB - Weak mutagenic activity was detected in several commercially available edible palm and corn oils using liquid incubation bioassays with Salmonella typhimurium TA1537. Chromatographic fractionation of unrefined palm oil established that mutagenic activity was present in three fractions that also contained fatty acyl hydroperoxides. Similar weak mutagenic activity was also demonstrated for linoleic and linolenic acid hydroperoxides. In all cases, the mutagenicity was abolished by exogenous catalase, implying that the observed activity was moderated by hydrogen peroxide. PMID- 2570469 TI - Caffeine toxicity is inversely related to DNA repair in simian virus 40 transformed xeroderma pigmentosum cells irradiated with ultraviolet light. AB - Human cells transformed by simian virus 40 (SV40) are more sensitive to killing by ultraviolet light when grown in caffeine after irradiation. The degree of sensitization at 2 mM caffeine (expressed as the ratio of the 37% survival dose for control cells divided by the 37% survival dose for cells grown in caffeine, i.e., the dose modification factor) was approximately 1.9 in transformed normal cells and 3.8-5.8 in excision-defective xeroderma pigmentosum (XP) groups A, C, and D cells. A large dose modification factor of 12 was observed in a transformed XP variant cell line. Chinese hamster ovary cells were not significantly different from transformed normal human cells, with a maximum dose modification factor of 1.5. Two radioresistant XP revertants that do not excise cyclobutane dimers gave different responses; one resembled its group A parent in being sensitized by caffeine, and one did not. These results can be interpreted on the basis of a single hypothesis that cells are killed as a result of attempts to replicate damaged DNA. Increased replication rates caused by transformation, increased numbers of replication forks in DNA caused by caffeine, and increased numbers of damaged sites ahead of replication forks in excision-defective cells are all processes that will consequently increase killing according to this hypothesis. A corollary is that the XP variant may be highly sensitized to caffeine because of excision defects at the DNA replication forks, an idea that may be important in designing cloning strategies for the XP variant gene. PMID- 2570470 TI - Comparison of DNA alkylation, fragmentation, and repair in maternal and fetal tissues of pregnant rats treated with a single dose of ethyl methanesulfonate, ethyl-N-nitrosourea, N-nitrosodiethylamine, and methyl-N-nitrosourea. AB - The occurrence and persistence of DNA damage, as detected by the alkaline elution technique, have been studied in some tissues of both fetal and adult Sprague Dawley rats (18th day of gestation) after administration of a single equimolar dose (0.5 mmol/kg) of ethyl methanesulfonate (EMS), N-ethyl-N-nitrosourea (ENU), N-nitrosodiethylamine (NDEA), and N-methyl-N-nitrosourea (MNU). EMS, ENU, and MNU, injected intravenously, produced a statistically significant increase of DNA elution rate, which is considered indicative of DNA fragmentation, in both maternal and fetal liver, kidney, and brain. NDEA, introduced by gastric gavage, induced DNA breaks in both liver and kidney of dams, but only in the liver of fetuses. The frequency of DNA lesions was found to vary with the four alkylating agents and in the three organs tested, to exhibit a different time course, and usually to be higher in maternal than in fetal tissues. Results provided by the concomitant determination of DNA binding levels demonstrated a satisfactory correlation with the amounts of DNA fragmentation. In contrast, the values of both these parameters did not show any positive correlation with the different susceptibility of the three organs to tumor induction. In conclusion, these findings suggest that when a compound is not available in radiolabeled form, measurement of DNA fragmentation may represent a useful alternative to the determination of DNA binding level in order to obtain information on the distribution of its reactive species in maternal and fetal tissues. PMID- 2570471 TI - Benzamide prevention of ultraviolet radiation-induced transformation as measured by anchorage-independent growth and the absence of correlation with thymidine dimer formation and DNA repair. AB - Synchronized human fibroblasts were exposed in early S phase to increasing doses of ultraviolet (UV) irradiation in the presence and absence of an antitransforming drug, benzamide. Cellular survival, initial thymidine dimer formation and its repair, and cellular phenotypic transformation were simultaneously monitored in the presence and absence of 1 mM externally added benzamide that reaches 8 to 15 microM intracellular levels. Cellular transformation as measured by an expression of anchorage-independent growth was inhibited by nontoxic doses of benzamide. Antitransforming action of benzamide is confined to low intracellular drug concentrations, which in the case of benzamide is in the 4-9 microM range. Because of the lack of effect of benzamide on the formation of UV-induced thymidine dimers and the specific repair of these dimers, these results suggest that the processes of thymidine dimer formation and its repair are not involved in the mode of action of benzamide that influences the expression of a transformed phenotype with low malignant vigor. PMID- 2570472 TI - Deletion screening at the hypoxanthine-guanine phosphoribosyltransferase locus in Chinese hamster cells using the polymerase chain reaction. AB - We have developed a rapid screening method using the polymerase chain reaction (PCR) for detecting deletion mutations at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus in Chinese hamster cells. DNA was extracted from spontaneous and ultraviolet (UV) light- and X-ray-induced hprt deficient mutants. Two primer sets were used to amplify 276 bp and 344 bp fragments containing the entire exon 3 and exon 9 coding sequence, respectively. The PCR was performed using Taq DNA polymerase for 40 cycles, and the PCR product was directly analyzed for the presence of the respective amplified DNA using electrophoresis on agarose gels stained with ethidium bromide. With this assay, we have analyzed 39 independently derived hprt-deficient mutants. Four of ten spontaneous mutants were found to have deletions in exon 9. UV light produced mutants with predominantly wild-type amplification patterns (10/14). X-ray induced mostly deletion patterns (11/15); six of these occurred only in exon 9, and five occurred in both exons 3 and 9. These observations are consistent with the classical notion that UV light induces predominantly missense mutations and X ray produces a high proportion of deletion mutations. Deletion mutations occurred most frequently at the 3' end of the hprt gene, suggesting the possible existence of hot spots for deletions in this region. The PCR assay for deletion detection has the advantage that it can be completed in less than 4 hr without using radioisotopes. This assay should be useful for routine deletion screening. PMID- 2570473 TI - Localization of insulin, glucagon and somatostatin in the pancreas of the adult newt, Notophthalmus viridescens. AB - The current study is designed to demonstrate the presence of immunoreactive insulin (IRI), glucagon and somatostatin in the adult pancreas. Methods include aldehyde fuchsin (AF) staining and peroxidase anti-peroxidase (PAP) immunochemical localization for light microscopy as well as protein A gold (PAG) staining for scanning electron microscopy (SEM) in conjunction with backscattered electron imaging (BEI). Our results show the presence of large clusters of AF positive cells within networks of highly vascularized pancreatic acinar tissue. PAP immunochemistry of pancreas serial sections exhibit positive immunoreactivity to the same AF-positive structure, thus demonstrating the presence of IRI. This immunoreactivity is found in a high percentage of cells in the islet-like structures. These cells tend to be centrally located within the cluster. Antibody specificity controls, including homologous antigen immunoabsorbance, as well as incubation of sections in normal guinea pig serum give negative immunoreactivity. Immunoreactive glucagon-containing cells and somatostatin-containing cells are distributed around the periphery of the central core of IRI-containing cells. SEM in conjunction with BEI confirm the presence of PAG within these cell clusters. We conclude that: (a) newt pancreatic IRI reacts in a specific manner with bovine antibody, suggesting a partial structural similarity to mammalian antigen; (b) IRI is localized within within pancreatic islet-like cell clusters and these IRI containing cells form a central mass which is surrounded by glucagon and somatostatin-containing cells; this cellular distribution is similar to that found in many mammals. PAG conjugated insulin antibody is detectable by SEM in conjunction with BEI in islet cells of the newt pancreas. PMID- 2570474 TI - Tracheal gland mucous cells stimulated in vitro with adrenergic and cholinergic drugs. AB - To determine the responsiveness of tracheal mucous cells to adrenergic and cholinergic stimulation, we analyzed changes in their structure induced by neurotransmitter-like agonists. Ferret tracheal rings were exposed for 30 min in vitro to one of the following: phenylephrine, isoproterenol, or bethanechol (all at 10(-5) M), in the presence of absence of appropriate antagonists. Electron microscopy and morphometric analysis revealed that the volume density of mucous cells (Vvmc, i.e. the space occupied by mucous cells in the submucosa) significantly decreased, and the surface density of mucous cell apical membrane (Svam) increased in response to isoproterenol and bethanechol but not to phenylephrine. In metabolic labeling experiments, the morphological changes were accompanied by secretagogue-evoked release of 35S-labeled macromolecules. Taken together, these data suggest that tracheal mucous cells secrete 35S-labeled macromolecules in response to beta-adrenergic and muscarinic agonists by an exocytotic process that involves a reduction in cell size. PMID- 2570475 TI - Mitosis comes apart. PMID- 2570477 TI - [Foot injuries caused by a chain saws]. AB - Two cases of chain saw injuries of the foot are reported. The protective footwear did not provide sufficient protection. Suggestions are made for improving the footwear and for compulsory approval of the safety equipment. PMID- 2570476 TI - [The effect of beta blockers combined with thiazides on serum electrolytes]. AB - The case reports of a total of 9,670 patients admitted to a medical department were reviewed manually. From this material, all patients receiving treatment with beta-blockers alone, bendroflumethiazide alone or combinations of these were selected. In addition, a control group was selected. The patients should have received treatment for at least seven days and they should not be "ill". Patients receiving treatment with beta-blockers had higher S-K+ (4.2 mmol/l) and lower standard bicarbonate levels (23 mmol/l) than matched controls. Patients receiving treatment with bendroflumethiazide had low S-K+ (3.6 mmol/l) and high standard bicarbonate (26 mmol/l). Patients receiving treatment with both bendroflumethiazide and beta-blocker had low S-K+ (3.4 mmol/l) and the same standard bicarbonate as patients who received bendroflumethiazide alone. A more limited group chosen according to stricter criteria confirmed the above mentioned results. The investigation demonstrated that treatment with both beta-blocker and bendroflumethiazide caused deterioration of the hypokalaemia induced by bendroflumethiazide despite the higher S-K+ during beta-blockade. This may be due to renal excretion of potassium during beta-blockade. PMID- 2570478 TI - Detection of fimbrial adhesins of ETEC using monoclonal antibody-based latex reagents. PMID- 2570479 TI - Spectrum of disease in macaque monkeys chronically infected with SIV/SMM. AB - Twelve rhesus and one pig-tailed macaque have been monitored for 28-41 months following experimental infection with 10(4) TCID of SIV/SMM. Twelve of the 13 animals became virus positive and seroconverted within 3 to 6 weeks of exposure; the remaining animal seroconverted at 6 months, but has remained virus negative. Six of the 13 animals (46%) died between 14 and 28 months post-infection, following prolonged clinical disease characterized by chronic diarrhea and weight loss, peripheral lymphadenopathy and hemogram abnormalities. Histologic findings ranged from prominent follicular hyperplasia to severe lymphoid depletion, with lymphoid tissues often showing an infiltrate of syncytial giant cells. One animal had intestinal cryptosporidiosis and two had brain lesions comparable to those seen in AIDS encephalopathy in humans. Three of the remaining seven animals have an ARC-like disease and are showing gradual deterioration of their clinical condition. These animals, as well as animals that died, had progressive decreases in CD4+ cells and CD4+/CD8+ cell ratios. These observations further document the marked clinical, pathologic and immunologic similarities between human AIDS and the SIV-infected macaque model. PMID- 2570480 TI - The role of bile acids in the reduction in lipopolysaccharide uptake by cultured rat Kupffer cells. AB - The influence of bile salts on the binding and uptake of Salmonella abortus equi lipopolysaccharide by cultured Kupffer cells was studied. In control preparations, the percentage of cell-associated lipopolysaccharide increased with time and reached a plateau after about 2 h incubation at 37 degrees C. About 1.2 micrograms lipopolysaccharide was associated with 10(6) Kupffer cells at this time interval. In the presence of 0.3, 0.6 and 1 mumol bile salts/ml the cell associated lipopolysaccharide was respectively, about 5%, 13% and 29% lower than in control cultures. In the presence of 1 mumol bile salts/ml, the association of lipopolysaccharide to cells at 0 degrees C was about 25% lower than in controls. Preincubation of Kupffer cells with 1 mumol bile salts/ml, with or without lipopolysaccharide, did not affect cell-associated lipopolysaccharide after removal of the bile salts. The rate of secretion of radioactivity by Kupffer cells was not influenced by the presence of bile salts during the uptake or the secretion periods. Bile acids proved to inactivate lipopolysaccharide. From these observations it was concluded that low concentrations of bile salts influence the binding and uptake of lipopolysaccharide by Kupffer cells. It was, therefore, considered likely that, in patients with obstructive jaundice, the high serum bile acid level accounts for spill-over of portal lipopolysaccharide into the systemic blood. PMID- 2570481 TI - Monocrotaline pneumotoxicity in mice. AB - Lung injury induced in rats by the pyrrolizidine alkaloid monocrotaline is a well documented model of pulmonary hypertension. To our knowledge, however, monocrotaline-induced cardiopulmonary injury has rarely been described and has never been quantitated in mice. In the present study, adult male mice received 2.4, 4.8, or 24.0 mg monocrotaline/kg body weight/day in the drinking water continuously for 6 weeks. These doses represent 1, 2, and 10 times the severely pneumotoxic regimen in rats. Pulmonary endothelial function was monitored by right lung angiotensin converting enzyme (ACE) activity, plasminogen activator (PLA) activity, and prostacyclin (PGI2) and thromboxane (TXA2) production. Light and electron microscopy were performed on the left lungs. Cardiac right ventricular hypertrophy was evaluated by the right ventricle to left ventricle plus septum weight ratio (RV/LV + S). Monocrotaline-treated mice exhibited a dose dependent decrease in lung ACE and PLA activities and an increase in PGI2 and TXA2 production, indicative of endothelial dysfunction. However, these responses were significant only after the highest monocrotaline dose. Light and electron microscopy revealed dose-dependent pulmonary inflammatory and exudative reactions. Unlike previous studies in rats, however, monocrotaline-treated mice developed relatively little lung fibrosis, cardiomegaly, or right ventricular hypertrophy, and no occlusive medial thickening of the pulmonary arteries, even at the highest dose level. These and previous data indicate that there are quantitative biochemical and qualitative morphological differences between mice and rats with respect to monocrotaline pneumotoxicity. Furthermore, in monocrotaline-treated mice (but not in rats) there appears to be a dissociation between lung endothelial dysfunction and inflammation on the one hand, and pulmonary hypertension and fibrosis on the other. PMID- 2570482 TI - New human colorectal carcinoma cell lines that secrete proteinase inhibitors in vitro. AB - Two new human cell lines, RCM-1 and CoCM-1, have been established from primary colorectal adenocarcinomas. Both cell lines were unique in that the cultures secreted trypsin inhibitors in vitro. The activities of these inhibitors were accumulated in serum-free media of both cell lines over a period of several days. Two inhibitors (PI-1 and PI-2) were isolated from serum-free conditioned medium in which RCM-1 was grown by anion-exchange and gel filtration high-performance liquid chromatography. PI-1 inhibited trypsin and chymotrypsin strongly, and pancreatic elastase weakly. Its molecular weight was about 57 kilodaltons (Kd) as determined by gel filtration chromatography. It cross-reacted with the antiserum elicited against human alpha 1-antitrypsin in double immunodiffusion. PI-1 corresponding to alpha 1-antitrypsin was also demonstrated immunohistochemically in both cell lines. PI-2 inhibited trypsin strongly, and chymotrypsin, kallikrein and plasmin weakly. It had higher molecular weight (200-300 Kd) than that of PI 1, and did not cross-react with antisera against human alpha 1-antitrypsin, alpha 2-macroglobulin, alpha 1-antichymotrypsin, alpha 2-plasmin inhibitor, inter-alpha trypsin inhibitor and urinary trypsin inhibitor. RCM-1 and CoCM-1 are the first colorectal adenocarcinoma cell lines that secrete functionally active trypsin inhibitors, including alpha 1-antitrypsin in vitro, and are useful for the study of tumor-cell derived proteinase inhibitors. PMID- 2570483 TI - Characterization of a human follicular thyroid carcinoma cell line (UCLA RO 82 W 1). AB - A thyroid tumor cell line has been established from the metastases of a follicular carcinoma in a female patient. Although the primary tumor released thyroglobulin (Tg) into the circulation (greater than 10,000 ng/ml), the uptake of I131 was less than 2%. After 37 replications the doubling time was 4 days and confluency was reached after 7 days from inoculation of 3 x 10(7) cells. This human thyroid tumor cell line has now been growing in culture for several years. An aneuploid chromosomal pattern was observed (62-82 chromosomes). A pair of X chromosomes was present but no Y chromosome was found which is compatible with the female origin of the cell line. EM studies revealed the presence of microvilli. Immunoperoxidase staining using specific anti-human Tg antisera indicated the presence of Tg within the cells. Nude mice developed solid-cystic tumors within 6 months after injection of the cells. The basal release of immunodetectable Tg, as measured in a perifusion system, increased in response to thyroid stimulating hormone (TSH) (P less than 0.025) or TSH combined with theophylline (P less than 0.001). Unusual isoenzyme patterns for galactose-1 phosphate-uridyltransferase (GALT) and phosphoglucomutase1 (PGM1) were detected in the tumor, compared with normal human fibroblasts and blood cells and isoenzyme patterns from the patient's lymphocytes. Because this malignant human thyroid follicular cell line has retained the ability to synthesize Tg it represents a valuable model for the study of human follicular carcinomas. PMID- 2570484 TI - Different contents of glycosaminoglycans in a human neoplastic salivary duct cell line and its subclone with a myoepithelial phenotype. AB - The glycosaminoglycans (GAG) biosynthesized by a neoplastic human salivary duct cell line, HSGc, and by its nontumorigenic subclone, HSGc-E1, having a myoepithelial-like phenotype, were examined by incorporation of [3H]-acetate into GAG. The rate of GAG radiolabeling in HSGc-E1 was significantly greater than that in HSGc. The radiolabeled GAG recovered from HSGc-E1 showed a distribution of 22 32% in the cells and 68-78% secreted into the medium, while the amounts of GAG in the cells and medium of HSGc were equal. Two-dimensional electrophoresis of GAG extracted from the cells demonstrated that HSGc-E1 contained a much greater amount of heparan sulfate (HS, 53.5% of total), while HSGc synthesized hyaluronic acid (HA, 17.5%), HS 38.8%, chondroitin sulfate (Ch-S, 27.6%) and dermatan sulfate (DS, 16.1%). Moreover, treatment of HSGc with sodium butyrate or dibutyryl cyclic AMP (each is a potent inducer of differentiation to myoepithelial-like cells) strongly enhanced GAG synthesis, while dexamethasone (an inducer of differentiation to a more functional duct epithelium) did not stimulate GAG synthesis. These findings suggest that biosynthetic changes in the GAG content of neoplastic salivary cells are associated with their myoepithelial differentiation. PMID- 2570485 TI - The morphology of the denuded epidermal basal cell layer of the hairless mouse after different preparation methods. A scanning and transmission electron microscopical study. AB - The denuded basal cell layer of the hairless mouse epidermis is described in the present scanning (SEM) and transmission electron microscopical (TEM) study. The suprabasal layers were removed mechanically after trypsinization or by extracellular calcium depletion. Trypsinization before removal of the suprabasal cells caused the basal cells to shrink. Characteristic surface plication and hemi desmosomal attachment to the basement membrane were generally preserved. SEM revealed partly maintained intercellular bridging, whereas by TEM such contacts were absent because half desmosomes were internalized. Total calcium depletion induced more serious damage to the basal cell surface, which was smooth with apparent perforations. However, cell bridges, and occasional desmosomes were present. The cell interior demonstrated important cellular injury. If the calcium deprived explants were allowed to recover in calcium-containing medium, the cells acquired an activated "regenerative" morphology, without junctions, similar to that observed in wound healing. Epidermal non-keratinocytes were seen only after trypsinization. Control experiments revealed that they adapted poorly to organ culture conditions. By TEM, we observed several interesting aspects of the differences, between dark and clear basal keratinocytes. This was unexpected because fixation studies had shown, that with the present fixation method, typical dark and clear cells do not occur in untreated epidermis. We believe that membrane injury through mechanical stripping of partly adhering epidermal layers induced "clear cells", whereby the neighboring cells appeared darker. This provides additional evidence as to the origin of the two sub-populations, dark and clear basal cells. The clear cells may be injured cells, caused by cell damage, and not by processes of cellular differentiation. The results of the present investigation supports the view that basal keratinocytes have a polygonal shape with numerous free surface extensions and they are anchored to the basement membrane with "foot pads". Our study also shows that SEM of the epidermal basal layer might be feasible. Various artifacts, however, must be considered, depending on the denudation method used. We prefer trypsinization to calcium depletion because it is less time-consuming and results in a cell morphology which in TEM is comparable to that of basal cells in untreated whole epidermis. Extra-cellular calcium depletion, however, might be useful as a method to prepare single cell suspensions for flow cytometry. Restoration of a normal calcium concentration after stripping, provides an opportunity to mimic wound healing in situ, as an alternative t PMID- 2570486 TI - Evaluation of human bladder tumors by ultrastructural morphometric techniques. AB - To estimate the malignancy of the human urinary bladder tumors, various cellular components of normal and neoplastic bladder epithelial cells were examined using morphometric techniques at the electron microscopic level. Nuclear to cytoplasmic ratios were found to be 1:5 in normal epithelial cells, 1:4 in superficial grade 1 (G1) tumor cells, 1:3 in superficial grade 2 (G2) tumor cells and 1:2 in invasive grade 3 (G3) tumor cells. Nuclear volumes were largest in G3 tumor cells, but no differences were seen in cell volumes between normal epithelial and tumor cells. The volumes and surface areas of the rough-surfaced endoplasmic reticulum (rER) and lysosomes per cell decreased progressively from G1 to G3 tumor cells. The volume density of tonofilaments was increased in G1, but decreased in G2 and G3 tumor cells. The cisternal thickness of the rER and Golgi complex was also increased in G1 tumor cells. From these results, it was concluded that morphometric analysis may be useful in evaluating the degree of differentiation and invasive potential of human bladder tumor cells in the low and intermediate risk groups. PMID- 2570487 TI - Establishment of a cell line producing basement membrane components from an adenoid cystic carcinoma of the human salivary gland. AB - A new cell line has been established from an adenoid cystic carcinoma arising in the submandibular gland of a 63-year-old woman. The cultured epithelial-like cells grew vigorously and adhered together to form a sheet. Immunohistochemical stainings for type IV collagen, laminin and fibronectin were clearly positive in the intercellular matrix and on the surface of the culture cells. Chondroitin 6 sulfate proteoglycan and heparan sulfate were also detected. Ultrastructural studies showed that the cells had abundant rough endoplasmic reticulum and a well developed Golgi apparatus. Rough endoplasmic reticulum near the cell surface was markedly dilated, and contained material of low electron density. This cell line would be useful for biological and biochemical studies on the mechanisms by which the stromal component is formed. PMID- 2570488 TI - Collecting duct origin of rat renal clear cell tumors. AB - The renal tubular segment from which clear cell tumors originate was investigated in the kidneys of rats treated with N-nitrosomorpholine. This tumor type, which in the rat closely resembles that in man, is made up of clear and granular acidophilic cells and arises from tubules lined by clear cells. The tubular origin of the tumors was established in serial sections by demonstrating connections between both clear cell tumors and tubules lined by clear cells, and renal tubules of normal appearance. In 45 clear cell lesions (17 tumors and 28 tubules) one or more such connections were identified which belonged to the collecting system. In accordance with their localisation in the kidney, the clear cell lesions were connected predominantly to tubules of the cortical collecting system and occasionally to outer medullary collecting ducts. As previously reported, oncocytic tubules and microoncocytomas were observed to originate from the same portions of the collecting system. Rarely, microadenomas and tubules consisting of both oncocytes and clear or granular acidophilic cells were also observed in the kidneys studied. PMID- 2570489 TI - Amplification of epidermal growth factor receptor (EGFR) gene and oncogenes in human gastric carcinomas. AB - DNAs from 37 human gastric carcinomas and seven lymph node metastases were analyzed for alterations of the epidermal growth factor receptor (EGFR) gene and oncogenes by the Southern blot hybridization method. The probes used were EGFR gene, c-Ha-ras, v-Ki-ras, N-ras, c-myc, v-myb, v-fos, c-erbB-2, v-erbA, v-abl and v-fes. Amplification of the EGFR gene was detected in only one poorly differentiated adenocarcinoma. Amplifications of c-myc gene and c-erbB-2 gene were each observed in two well differentiated adenocarcinomas. One of these tumors had coamplification of c-erbB-2 and c-erbA genes but there were no amplifications nor rearrangements of other oncogenes. The poorly differentiated adenocarcinom with amplified EGFR gene also showed enhanced expression of EGFR gene by Northern blot analysis and additionally had strong synchronous immunoreactivity for EGFR and EGF. PMID- 2570490 TI - Alpha-smooth muscle actin is expressed in a subset of bone marrow stromal cells in normal and pathological conditions. AB - A series of 217 trephine bone marrow biopsies from adult patients and specimens from 16 fetuses and 5 infants were examined for the presence of stromal myoid cells (MCs) using a monoclonal antibody recognizing alpha-smooth muscle actin. In the normal adult bone marrow, stromal cells did not contain alpha-smooth muscle actin, whereas during fetal life, many alpha-smooth muscle actin-containing MCs were connected with vascular sinusoids in the primitive bone marrow. This cell type reappeared in various characteristic distribution patterns in adult bone marrow during different neoplastic and non-neoplastic conditions including metastatic carcinoma, Hodgkin's disease, multiple myeloma, hairy cell leukemia, acute myeloid leukemia (FAB M4, 5, 7) and chronic myelo-proliferative diseases. In general, the appearance of MCs was associated with a slight to pronounced increase in the deposition of reticulin and collagen fibers. We propose that bone marrow MCs represent a distinct subpopulation of fiber-associated or adventitial reticular cells undergoing cytoskeletal remodeling in response to various stimuli. PMID- 2570491 TI - Cytotoxic action of the lysosomotropic agent L-leucine methyl ester on mammalian skeletal muscle. The role of the sarcoplasmic reticulum in producing myofilament damage. AB - Incubation in vitro of mouse or rat diaphragms or mouse soleus muscle with the lysosomotropic agent leucine methyl ester (10 mM) produced a slowly-developing swelling of the sarcoplasmic reticulum (SR), showing that this organelle is able to take up amino acid methyl esters and conforming with previous suggestions that the SR may serve some of the functions of the lysosomal system in muscle cells. Cellular damage followed, sometimes associated with the shrinkage of the SR, but experiments with inhibitors of lysosomal cathepsins suggest that acid hydrolases were not implicated in this damage. It is suggested that the system producing myofilament damage is located on the SR and that it may be directly activated by membrane perturbation. PMID- 2570492 TI - The effect of pinealectomy and jejunal loop diversion on rat small bowel crypt cell kinetics. AB - Previously, it was found that diversion of an isolated loop of jejunum into the colon was associated with a significantly diminished crypt cell proliferation rate in the isolated loop, probably principally because of the diminished amount of nutrient available to the diverted mucosa. It has also been shown previously that removal of the pineal gland is associated with a considerable increase in the jejunal crypt cell mitotic rate. In the present investigation it was found that following pinealectomy, whilst the rise in crypt cell proliferation elsewhere in the rat small intestine was maintained at the expected level, the mitotic rate in the crypts of an isolated jejunal loop attached to the colon was also increased to a similar level, despite the fact that this isolated loop was in contact with a considerably diminished level of luminal nutrients. Thus, the expected hypoproliferative effects of jejunal isolation were overridden by the hyperproliferative effects of pinealectomy and the effects of pinealectomy appeared to be independent of the particular changes in luminal environment produced in this experiment. The significance of these findings is discussed in the light of the available literature. PMID- 2570493 TI - [Fat embolism in severe trauma]. PMID- 2570494 TI - Pregnancy and inflammatory bowel disease. AB - Conclusions about the relationship between the pathophysiology and treatment of inflammatory bowel disease and the physiology and management of pregnancy are based on the results of several large physician surveys and retrospective chart reviews. Patients with active disease fare worse than those with inactive disease. There is little evidence that pregnancy affects the course of inflammatory bowel disease or that inactive inflammatory bowel disease affects the course of pregnancy. Judicious medical therapy is effective in controlling inflammatory bowel disease during pregnancy. Sulfasalazine or steroid therapy should not be withdrawn in a patient who needs it to achieve or maintain a quiescent state of inflammatory bowel disease during the course of pregnancy. Immunosuppressive therapy should be avoided. Aggressive medical therapy with total parenteral nutrition in a team approach with a gastroenterologist, surgeon, and perinatologist usually avoids the need for surgical intervention during pregnancy with a good fetal outcome in a patient whose disease is active. Contraception against pregnancy need only be considered in those patients whose disease is so severe that operative therapy is imminent. PMID- 2570495 TI - [Drug therapy of delusional parasitosis. The importance of differential diagnosis for psychopharmacologic treatment of patients with delusional parasitosis]. AB - In 34 patients suffering from delusional parasitosis the relevance of an accurate differential diagnosis with respect to pharmacological treatment was investigated. Under a psychopharmacological therapy of the delusion's additional psychiatric symptomatology in 17 patients (50%) a full remission and in other 5 patients an improvement of the delusional symptoms was observed. The unexpected high recovery-rate is explained by the fact that all of the patients with additional depression showed a reduction also of the delusional symptomatology after a treatment with antidepressants. In contrast to that, patients with organic brain syndromes underlying the delusions had a significant worse outcome, which seems to be caused by the considerable lack of effective psychopharmacological treatment of such states. The results indicate that an accurate psychiatric differential diagnosis is of great importance for the psychopharmacological therapy of patients suffering from delusional parasitosis. In this context the Viennese decision-tree for delusional syndromes is discussed. PMID- 2570496 TI - [Therapy and compliance in angina pectoris--analysis of a physician and patient survey]. AB - Positive compliance is essential for the successful treatment of coronary heart disease. To study the reasons for non-compliance of patients with angina pectoris 50 patients out of a sample of 2000 persons were recruited. Randomized telephone calls were used. The 50 angina pectoris patients were interviewed using a semi structured questionnaire. Another questionnaire was sent to all general practitioners and specialists for internal diseases in Austria. 675 doctors replied. The responding physicians reported a 68.9% positive compliance of the patients to pharmaceutical treatment of angina pectoris. Nevertheless most of the physicians believe that about half of the patients sometimes forget to take their medicine or change their therapeutic scheme or don't take the prescribed drugs if they have no symptoms. Patients suffering from angina pectoris confirm compliance problems, but significantly less often than doctors. PMID- 2570497 TI - Mechanisms of increased sensitivity for detection of thyroid stimulating antibodies by use of hypotonic medium. AB - We have attempted to clarify the mechanism whereby the sensitivity to detect thyroid stimulating antibodies by use of cultured thyroid cells is enhanced when sodium chloride is removed from isotonic medium. Preexposure of cultured porcine thyroid cells to 10(-4) mol/l cycloheximide increased the subsequent cAMP response to TSab stimulation in isotonic NaCl medium, but not in hypotonic NaCl free medium. Crude membrane fractions from porcine thyroid tissues were incubated with 6 Graves' IgGs in NaCl or NaCl-free medium at 37 degrees C for 1 h, centrifuged, washed, added [125I]TSH, and incubated for another hour. The result of the 'residual TBII assay' indicated that all 6 IgGs exhibited greater inhibition of [125I]TSH binding to the membranes when pre-incubated under hypotonic than isotonic conditions. When cultured porcine thyroid cells were incubated with 7 TSab samples in NaCl or NaCl-free medium depleted with 3 isobutyl-1-methylxanthine at 37 degrees C for 1 h, washed, added the NaCl-free medium containing 3-isobutyl-1-methylxanthine, and incubated for another 2 h, the use of NaCl-free medium during the pre-incubation resulted in greater increase in cAMP elicited by all 7 samples. In summary, 1) an inhibitory mechanism upon adenylate cyclase seems to be involved under isotonic conditions and may require new protein synthesis; 2) the binding of TSH receptor antibodies to the TSH receptor increases when NaCl is removed from isotonic medium, and 3) these factors are considered responsible for the enhanced sensitivity under hypotonic conditions. PMID- 2570498 TI - Lesions and repair of cells of maternal mice after valproic acid (VPA) treatment on day 8 of pregnancy: an enzyme histochemical analysis. AB - Mice received a single teratogenic dose of the antiepileptic drug valproic acid (VPA; 500 mg/kg i.p.) on 8th d of pregnancy. The effects of VPA were studied primarily by enzyme histochemical means over a period of 48 h. Plasma membrane associated hydrolases were more affected than lysosome-associated ones. Lesions were not found in the spleen, but a slight response, without morphological damages, was observed in the lung and thymus. Comparatively more severe injuries, which were not accompanied by clear-cut structural changes either, occurred in the liver. The most severe damages including morphological lesions were noticed especially in the proximal tubules of the kidney. The first damages were already seen after 3 h and concerned primarily the liver. After 12 h, a maximal response to VPA was detected in all organs. Repair of most of the enzymatic lesions set in after 24 h. After 48 h, lesions were not longer seen in the lung and thymus. In contrast, enzymatically and morphologically injured proximal renal tubules could still be found after this period as a response to a single dose of VPA; in the liver only enzymatic lesions were present at this stage. PMID- 2570499 TI - Gamma-glutamyltransferase induction by glucocorticoids in rat liver: age dependence, time-dependence, dose-dependence, and intralobular distribution. AB - Postnatal responsiveness of rat-liver gamma-glutamyltransferase (GGT) to glucocorticoids (GC) has been defined by investigating: age-dependence, time dependence, hormonal dose-dependence, and lag-time of the enzyme re-expression; half-life of the induced enzyme activity; dynamics of the enzyme reappearance in the liver tissue. Hydrocortisone-acetate (HC) or dexamethasone (DEX) were administered to the animals starting 1, 2, 3, 4, or 5 d before killing, at the doses of 25 micrograms or 1 microgram/(g b. w. x d), respectively. In 14 d old rats, after a lag-time of about 20 h (DEX) or 30 h (HC), GGT activity progressively increased up to 38 and 31 times the control value, respectively, at 5th d; the enzyme re-expression was linearly hormone dose-dependent; half-life of the induced enzyme activity was about 36 h. In 21 d old rats, GGT re-induction behaved as in 14 d old animals, except that the induced activity was about half that of each correspondent treatment. In 28 d old rats, a very low but significant GGT activity was re-expressed only after hormonal treatments longer than 48 h. In 35 and 77 d old rats, significant GGT activity was never re induced. GGT was re-expressed in liver parenchyma, with a defined space-course. In 14 d old rats, GGT reappeared first in periportal areas, then in acinar zone 1, finally in acinar zone 2. While the animals were ageing, GGT re-expression occurred to lesser and lesser extents in liver tissues, because of a progressive space-restriction from acinar zones 1 and 2 to zone 1 and finally, in 35 d old rats, to periportal areas. In adults, GGT was re-expressed only by rare hepatocytes in periportal spaces. Acinar zone-3 hepatocytes did never re-express GGT, irrespectively of the animal age. Thus, 2 rat hepatocyte populations could be distinguished (1 responsive, the other unresponsive to GC for GGT re expression), the relative proportion of which changes in favour of the unresponsive one while the animal ages. Hepatic GGT re-induction by GC, occurring after a long lag-time, does not follow the typical model of hormonal induction. Previous permissive cell changes seem to be required. Hepatocyte-GGT re expression by GC appears to be inversely correlated with the differentiation level and the cytochrome P-450 amount (activity) of the cell as limiting factors for the triggering of the enzyme induction. PMID- 2570500 TI - A comparative study of the orbicularis oculi muscle to the adductor pollicis muscle during vecuronium neuromuscular blockade. AB - This study compared the orbicularis oculi muscle to the adductor pollicis muscle during vecuronium blockade. The orbicularis oculi muscle was found to be more resistant to and recovered from vecuronium neuromuscular blockade faster than the adductor pollicis muscle. Thus, caution should be utilized when assessing neuromuscular block with the orbicularis oculi muscle. PMID- 2570502 TI - Amphetamine stereotypy in cats and neurotransmitter interactions in the caudate nucleus. I. Effects of intracaudate injections of haloperidol, GABA and bicuculline. AB - Intracaudate (into the rostrodorsal part of the caudate nucleus) injection of haloperidol in free moving cats did not modify spontaneous or d-amphetamine induced stereotyped behavior, while administered intraperitoneally, haloperidol blocked the stereotypy. These results, in accordance with some views, suggest that in this region of the caudate nucleus, the dopamine inhibitory receptors prevail over the excitatory ones. The effects of intracaudate bicuculline (decrease in the number of all stereotyped movements, increase in the number of ipsilateral amphetamine-induced or spontaneous head movements over the contralateral ones) were more pronounced than those of intracaudate GABA (increase only in the number of nonfocused lickings). The present results support the view that interactions between dopamine and GABA occur in the caudate nucleus in cats during amphetamine stereotypy. PMID- 2570501 TI - [Effect of Chinoin-103 on Na+, K+-activated adenosine triphosphatase of the rat heart]. AB - Effect of a new beta-blocking agent, a cardioselective aryl oxybutanolamine derivative, the Chinoin-103 on basal and total ATP-aze activities has been studied in total homogenizatum of rat heart. The effect has been compared to previous results of propranolol and to effect of practolol, respectively. It has been established that DL-Chinoin-103-similarly to DL-propranolol but in a little higher concentration--has significantly impeted both basal and total ATP-aze activities. Since in the nase of practolol similar effect was not obsreved and furthermore the effect could not be suspended by isoproterenol, the authors suppose that impediment of sarcolemmal ATP-aze activities cannot primarily be attributed to beta-blocking effect of compounds. Study of effect of racemic Chinoin-103 on enzymatic kinetic parameters of basal and total ATP-aze acticities has shown that primarily the reaction rate had decreased, affinity to substratum had not changed in the case of total ATP-aze and it has moderately decreased in the case of basal ATP-aze. The results of this publication has drawn attention to the fact that some beta-receptor blocking compounds may have other specific membrane effects besides antagonism on beta-receptors. PMID- 2570503 TI - Effects of a ganglionic blocking agent on the accumulation of DOPA in peripheral organs. PMID- 2570504 TI - Neurogenic contractions in the tail artery of normotensive rats are mediated by noradrenaline and ATP via post-junctional alpha 1- and alpha 2-adrenoceptors and P2x-purinoceptors. PMID- 2570505 TI - Cerebrovascular serotonergic receptors mediating vasoconstriction: further evidence for the existence of 5-HT2 receptors in rat and 5-HT1-like receptors in guinea-pig basilar arteries. AB - Pharmacological experiments were carried out on isolated basilar arteries (BA) from the brain vasculature of guinea-pig and rat in order to characterize post junctional serotonergic receptors mediating contraction by the use of selective agonists and antagonists. The sensitivity to 5-HT was higher, but the intrinsic activity lower, in guinea-pig compared to rat vessels. The contractile potency of the 5-HT1 agonists, 5-carboxamidotryptamine (5-CT), was three times higher than 5 HT in guinea-pig but 16 times lower in rat BA. In arteries from both species the 5-HT1A agonist, 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT), only caused weak contraction. In rat BA, where the serotonergic contractile receptors are ketanserin-sensitive, mesulergine inhibited the contraction in doses high enough to block 5-HT2 receptors, and also propranolol slightly inhibited the contraction, probably due to its binding to these receptors. Methiothepin, a potent antagonist of the 5-HT1-like receptors, affected the contraction in a non competitive manner. The antagonist profile was different in guinea-pig BA: propranolol was ineffective, mesulergine caused a slight, non-surmountable inhibition, whereas methiothepin acted as a true, competitive antagonist. The data support previous suggestions that the serotonergic contraction in rat BA is mediated by 5-HT2 receptors, whereas the present data show that 5-HT1-like receptors predominate in guinea-pig BA. PMID- 2570506 TI - Transfer of DOPA from the sympatho-adrenal system to the pancreas, liver and kidney via the blood circulation. AB - The concentration of 3,4-dihydroxyphenylalanine (DOPA) was low in the pancreas, liver, kidney, spleen, salivary glands, heart and adrenal glands of untreated mice, but increased following inhibition of DOPA decarboxylase by 3 hydroxybenzylhydrazine. The ratio of the accumulation of DOPA to the concentration of noradrenaline (i.e. the density of sympathetic nerves) was greater in the kidney, liver and, particularly, pancreas than in the other organs studied, suggesting that DOPA occurred outside sympathetic nerves in these organs. The tyrosine hydroxylase inhibitor alpha-methyltyrosine almost completely inhibited the accumulation of DOPA in all organs. The DOPA accumulation was enhanced in all organs by the alpha-adrenoceptor antagonists phentolamine and yohimbine. The results indicate that the DOPA was formed in the sympatho-adrenal system. In the pancreas, liver and kidney, most of the DOPA accumulated might have been formed outside these organs and transferred there via the bloodstream. PMID- 2570508 TI - Genetic relatedness of the kemerovo serogroup viruses: II. RNA-RNA blot hybridization and gene reassortment in vitro of the Great Island serocomplex. AB - The majority of the thirty-two Great Island serocomplex isolates examined exhibit distinct dsRNA polyacrylamide gel profiles. Yet, these viruses are closely related by blot hybridization with only two genes showing significant sequence divergence. Gene reassortment was demonstrated between selected pairs of the Great Island serocomplex viruses with two different geographic regions represented. The majority of the reassortant progeny from the cross of selected pairs resulted in progeny with multiple gene-replacements. The ability of these selected isolates to reassort confirms the close taxonomic relationship of the isolates in spite of their geographic distribution. PMID- 2570507 TI - Mechanism of altered cytoskeleton organization in influenza virus infection. AB - We followed the autophosphorylation of cytoskeleton (CS) isolated from control chick embryo cell membranes (CS-C) and from these membranes after influenza virus adsorption (CS-V) under conditions allowing to determine the activity of a single type proteinkinase. The Ca2+ dependent calmodulin (CaM) kinase used different substrates from CS-V than did the c'AMP dependent proteinkinase. The catalytic subunit (c-subunit) of the c'AMP dependent proteinkinase added from outside phosphorylated the same polypeptides than the endogenous c'AMP dependent proteinkinase, the further being more active than the latter. The purified influenza virus incorporated 32P in the presence of the c-subunit only. Incubation of influenza virus with the c-subunit caused morphological changes visible by electron microscopy. The pleomorphy of the particles as well as their electron transmissibility were enhanced in result of structural alterations and rarefaction of surface spikes of the haemagglutinin and neuraminidase. The contractibility of CS isolated from normal CEC and of the CS from CEC by 15 min postinfection (p.i.) was determined according to the actomyosin ATPase activity. The ATPase activity of the cytoskeleton in the presence of the Ca2+/CaM and that in the presence of c'AMP were used as controls. The virus as well as the Ca2+/CaM increased the ATPase activity. EGTA had no effect but did not interfere with virus stimulation, while c'AMP blocked the virus-induced enhancement of the ATPase activity. PMID- 2570509 TI - Genetic relatedness of the Kemerovo serogroup viruses: III. RNA-RNA blot hybridization and gene reassortment in vitro of the Chenuda serocomplex. AB - The dsRNA polyacrylamide gel profiles of five Chenuda serocomplex viruses were distinct. Blot hybridization and gene reassortment in vitro studies demonstrated that the Chenuda serocomplex may be divided into three sets: Chenuda, Huacho, and Mono Lake. Genes were highly conserved among members of each set, whereas genes were not highly conserved between members of different sets. Gene reassortment was demonstrated in intra-set crosses, but inter-set crosses did not yield reassortant progeny. The taxonomic significance of these data to the Chenuda serocomplex is discussed. PMID- 2570510 TI - Modulation in response to temperature of Mayaro virus proteosynthesis in Aedes Albopictus cells. AB - Incubation of Aedes albopictus cells infected with Mayaro virus at 37 degrees C causes inhibition of virus replication. During the first hour post infection (p.i.) incubation at 37 degrees C inhibited cellular and virus proteosynthesis. A preferential translation of heat shock proteins 82 kD and 70 kD was observed. After incubations longer than 1 hr at 37 degrees C, a switch to normal pattern of cell protein synthesis occurred without recovery of virus proteosynthesis. In addition, preferential synthesis of three major virus proteins of 62 kD, 50 kD and 34 kD was observed, when infected cells incubated at 37 degrees C were shifted down to 28 degrees C. PMID- 2570511 TI - Monkey pox in humans: current results. AB - The presented studies were performed at the WHO Collaboration Center for Smallpox in Moscow in the framework of the WHO monkey pox project. The authors recommend improved methods for rapid detection of orthopoxvirus antigen, namely passive haemagglutination (PHA) using dried stable red blood cells and ELISA in order to provide more rapid and efficient laboratory diagnosis under field conditions. Independent serologic diagnosis of monkey pox by ELISA-adsorption (ELISA-A) was proved of value for epidemiological studies and for detection of inapparent infections. The application range of the latter technique and its limitations were also determined. PMID- 2570513 TI - The role of suppressive macrophages in influenza virus-induced immunosuppression. AB - To confirm the role of macrophages in influenza virus (IV)-induced immunosuppression (IS) of mice, silica, a selectively destructive agent for macrophages, was tested for its ability to influence the IV-induced IS. Silica completely abolished that effect. In contrast, Indomethacin--a prostaglandin inhibitor--did not influence the IV-induced IS. Peritoneal macrophages of mice infected with IV were suppressive for plaque forming cell (PFC) response of normal spleen cells in vitro. The assumption was made that suppressive macrophages were induced by IV in mice. However, no suppressive factors were found in the culture fluid of these cells. PMID- 2570512 TI - Monoclonal antibodies neutralizing human leukocyte acid- and thermolabile interferon alpha. AB - Using a high titred human leukocyte IFN alpha preparation which contained both acid- and thermolabile (AL-IFN alpha) and acid- and thermostable IFN alpha species in 9:1 proportion for immunization of BALB/c mice, five hybridomas secreting monoclonal antibodies that reacted with AL-IFN alpha were obtained. In antiviral and antiproliferative tests on HL-60 cells, their products showed high degree of specificity for AL-IFN alpha. The results suggest that both the "normal" leukocyte AL-IFN alpha and the IFN alpha found in sera of autoimmune and other chronic patients might belong to the same subtype of IFN alpha. PMID- 2570514 TI - Erve virus, a probable member of Bunyaviridae family isolated from shrews (Crocidura russula) in France. AB - An apparently new agent, provisionally named Erve virus, was isolated in 1982 from tissues of three white toothed shrews, Crocidura russula, trapped near Saulges village in Western France. Results of virological and ultrastructural studies suggest that this virus belongs to the Bunyaviridae family and is a Bunyavirus-like agent. Serosurveys indicate that Erve virus had apparently a large geographical distribution in France and infects rodents, insectivores, wild boars (Sus scrofa), red deer (Cervus elaphus), sheep, herring gulls (Larus argentatus) and humans. Blood donors living in the vicinity of the Saulges area exhibit the highest incidence of antibody against Erve virus. PMID- 2570515 TI - Red plaque formation of Coxiella burnetii and reduction assay by monoclonal antibodies. AB - A red plaque technique for C. burnetii which utilizes primary chicken embryo cells, is described. Red plaques could be consistently detected as early as 6 days, usually 8 days post inoculation (p.i.), reflecting that C. burnetii proliferated within the phagolysosomes of host cells. Incubation with phase II monoclonal antibodies or inactivated immune sera containing phase I and phase II antibodies or phase II antibodies only, markedly reduced phase II C. burnetii red plaques. On the other hand, red plaques from phase I organisms increased several times when phase I cells were mixed with phase I monoclonal antibodies or inactivated immune sera containing phase I and phase II antibodies. By indirect red plaque reduction assay red plaque production by phase II cells could be reduced as well. PMID- 2570516 TI - Detection of swine pox and buffalo pox viruses in cell culture using a protein A horseradish peroxidase conjugate. AB - Buffalo pox virus antigen was detected in Vero cells and swine pox virus antigen in the cytoplasm and nucleus of PK-15 and IB-RS-2 cells as early as 6 hr post infection (p.i.) by indirect immunoperoxidase technique using a Protein A horseradish peroxidase (HRP) conjugate. The viral antigens localized in the cytoplasm of infected cells were the most prominent after 24 hr p.i. PMID- 2570517 TI - Enhanced full-length transcription of Sindbis virus RNA by effective denaturation with methylmercury hydroxide. AB - 49-S Sindbis virus RNA was reverse transcribed into a complementary DNA. The RNA templates were denatured by three different methods prior to DNA synthesis. Efficient full-length transcription was only achieved after treatment with methylmercury hydroxide. PMID- 2570518 TI - Society of Uroradiology Award paper. The pars infravaginalis gubernaculi: importance in the identification of the undescended testis. AB - The gubernaculum, a cordlike structure that extends from the testis to the scrotum and guides the testis in its descent, has a bulbous termination, the pars infravaginalis gubernaculi. We reviewed seven surgically proved cases in which the pars infravaginalis gubernaculi was mistaken for an undescended testis on imaging studies. The studies included sonography (five cases), CT (two cases), and MR imaging (two cases). Identification of the mediastinum testis on sonograms or MR images confirms that a structure is testis and not the pars infravaginalis gubernaculi. In two of these patients, testicular venography was used to further identify the undescended testis. In these two patients, the pampiniform plexus, which is used to identify the presence and position of testis, was located adjacent to the pars infravaginalis gubernaculi. Our experience indicates that the pars infravaginalis gubernaculi can be similar in appearance to the undescended testis on any imaging study. The finding of an apparent cord leading to a testislike structure on caudad sectional imaging does not obviate searching for the testis as far as the renal hila. PMID- 2570519 TI - Is "stunned myocardium" a protective mechanism? Effect of acute recruitment and acute beta-blockade on recovery of contractile function and high-energy phosphate stores at 1 day post-reperfusion. AB - There is little doubt that the "stunned myocardium" is amenable to therapeutic intervention, as a host of diverse pharmacologic agents have all been shown to improve short-term contractile function of viable, previously ischemic myocardium. However, few studies have addressed the question: Should the stunned myocardium be forced to contract? If the stunned myocardium is a protective mechanism, then acute recruitment could have later deleterious consequences on recovery of contractile function and high-energy phosphate stores. Conversely, acutely "resting" the heart (i.e. by beta-adrenergic blockade) could conceivably enhance or accelerate recovery of the stunned, postischemic tissue. We therefore sought to assess the immediate and longer-term effects of acute recruitment and acute beta-blockade on regional wall thickening (WT: using two-dimensional echocardiography) and adenosine triphosphate (ATP) content in the canine model of the stunned myocardium. Anesthetized open-chest dogs underwent 15 minutes of transient coronary artery occlusion. At 30 minutes following reperfusion, the dogs acutely received either: the ultrashort-acting beta-blocker esmolol, the afterload reducing and cardiostimulatory agent hydralazine, or saline. As anticipated, hydralazine enhanced contractile function of the stunned tissue in the short term: WT at 2 hours after treatment was 53.7 +/- 6.9% versus 7.1 +/- 6.5% in treated versus saline controls (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570520 TI - Multifocal atrial tachycardia: mechanisms, clinical correlates, and treatment. AB - MAT is an uncommon arrhythmia most often seen in elderly patients with chronic pulmonary disease who are critically ill due to acute respiratory or cardiac decompensation. Its importance lies in the fact that it is commonly mistaken for AF, since both disorders are characterized by narrow ventricular complexes, irregular rates, and (depending on the ECG lead observed in MAT) by an apparent lack of P wave activity. This may lead to treatment with digoxin, a drug known to be ineffective in the therapy of MAT, with the potential for producing toxicity in patients who are predisposed. The incidence of MAT in hospitalized patients in various studies ranges from 0.13% to 0.40%. The mechanism of the arrhythmia is thought to be triggered activity arising from increased intracellular calcium stores that may be produced by hypokalemia, hypoxia, acidemia, and increased catecholamines, characteristics commonly found in patients with MAT. COPD, coronary artery disease, CHF, and infection (both pulmonary and nonpulmonary) are the most common clinical settings of MAT. Mortality is very high in all patients studied, ranging from 38% to 62%, and is due to their underlying disease processes and not to the arrhythmia. The need for intubation and mechanically assisted ventilation portends a particularly poor prognosis for survival. Treatment should initially consist of correction of the precipitating causes, as it is common for patients to convert to sinus rhythm both spontaneously and after these measures are taken.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570521 TI - The relationship of the sympathetic nervous system and the renin-angiotensin system in congestive heart failure. AB - Congestive heart failure is a complex clinical syndrome characterized by circulatory and metabolic abnormalities. It has been apparent for more than 25 years that the sympathetic nervous system and the renin-angiotensin-aldosterone system are markedly activated in the late stages of heart failure. These two systems interact to facilitate sympathetic drive and promote salt and water retention. Circumstantial evidence is now accumulating to indicate that excessive sympathetic drive and angiotensin II activity may contribute to the pathophysiology of heart failure. These observations suggest that a dual strategy of modulating sympathetic nervous system activity to the heart while blocking angiotensin II activity may provide a rational therapeutic approach to the treatment of heart failure. Xamoterol, a beta 1 partial agonist, may enhance myocardial contractile force in the steady state, while acting to inhibit excessive sympathetic drive during exercise or severe heart failure. The concomitant use of a converting-enzyme inhibitor would be expected to blunt the detrimental effects of excessive angiotensin II activity. Modulation of adrenergic drive coupled with inhibition of marked angiotensin II activity may be potentially more effective in the treatment of congestive heart failure than either strategy used alone. PMID- 2570522 TI - Physiologic relation between cardiac cycle and QT duration in healthy volunteers. AB - To determine the relation between QT duration and RR interval on the electrocardiogram, 2 studies were undertaken in 23 young healthy volunteers (mean age 24 years). In study 1, the electrocardiogram (paper speed at 50 mm/s, 800 measurements/subject) was recorded in 11 subjects (5 men, 6 women) at rest, during dynamic exercise and after rapid intravenous injections of isoproterenol (0.2 to 12.8 micrograms), before and after intravenous atropine (2 mg). In study 2, the QT-RR relation was studied at rest and during exercise in 12 subjects (6 men, 6 women) before and after oral propranolol (80 mg). The results confirmed a monoexponential individual relation of the QT and RR intervals during rest and exercise: QT = A-B.Exp (-k.RR). By pooling the RR-QT plots from the 11 subjects in study 1, we found that the measured QT interval = 425-676.Exp (-3.7.10(-3) RR. During isoproterenol-induced tachycardia, QT either did not change or increased and this may indicate an increase in temporal dispersion of ventricular repolarization. Atropine-induced tachycardia produced changes similar to those resulting from exercise testing. Propranolol did not change the QT-RR relation despite a lengthening in RR intervals. These results suggest that physiologic QT RR adaptation is mainly under parasympathetic control. PMID- 2570523 TI - Use of colloidal gold cytochemistry in the study of the basic cell biology of cancer. AB - We are currently investigating the morphologic aspects of two areas of the basic cell biology of cancer: tumor-specific surface antigens as targets for immunotoxins, and the phenomenon of multidrug resistance in chemotherapy of human tumors. Colloidal gold cytochemistry has provided a useful method for the electron-microscopic cytochemical detection of materials endocytosed by cells in culture. This technique has been used to study the internalization pathway of ligands bound to the surface of cancer cells, particularly antibodies for use as immunologic targeting reagents for the construction of immunotoxins. These colloidal gold conjugates with monoclonal antibodies have demonstrated the internalization of these immunologic reagents through coated pits and receptosomes, which is a necessary step in the delivery of immunotoxins into the cell where they can mediate their cell-killing functions. Morphologic methods have been employed for the screening and selection of monoclonal antibodies reactive with the surface of human ovarian cancer cells for use as immunotoxins and have demonstrated the in vivo activity of immunotoxins made with these antibodies and Pseudomonas exotoxin in a nude mouse model system. In other studies, we have employed such reagents for the immunocytochemical detection of the surface expression of P170, the cell-surface efflux pump protein responsible for the phenotype of multidrug resistance in tumor cells, and to investigate the distribution of this protein by using immunocytochemistry in normal human tissues. These results have suggested a role for P170 in normal cell membrane transport of metabolites in various organ systems. PMID- 2570524 TI - Brain iron: a lesson from animal models. AB - Brain and blood iron deficiency (ID) can be nutritionally induced. Significant behavioral and brain-biochemical changes are observed in rats rendered iron deficient, including complete reversal of the circadian cycles of motor activity, changes in thermoregulation and stereotyped behavior, and an increased pain threshold. The increase in pain threshold is affected by diurnal factors and peripheral treatment with beta-endorphin has a significant analgesic effect, implicating selective changes in the blood-brain barrier. These effects along with modifications in responses to dopaminergic drugs, interactions of ID with neuroleptic drugs, and modifications in behavior as a result of selective brain lesions, lead to two conclusions: this animal model is appropriate for human anemia and the best explanation for the variety of behavioral and brain biochemical changes in ID rats is that the principal effect of brain ID is a selective decrease in the functional level of the dopaminergic D2 system. PMID- 2570525 TI - Fungal colonization of the esophagus. AB - A prospective study was conducted in 224 patients to determine the clinical significance of esophageal colonization with yeasts under different conditions. In accordance with the results of direct smear microscopic examination and culture of esophageal brushings, patients were divided into three groups: positive, negative, and the patients, in whom saprophytic forms were detected. A higher prevalence of positive findings was noted in patients with predisposing factors for yeast invasion than in patients free of underlying disease. Eleven percent of patients with an endoscopically normal appearing esophagus were positive. We have the impression that this situation may represent a preclinical condition of fungal esophagitis. Patients treated with H2-blocking agents showed a significantly higher incidence of positive findings, than did those without such treatment. Whether patients suffering from a refluxesophagitis resistant to long-term treatment with H2 blockers, but with a significant colonization by yeasts, could benefit by an additional treatment with antimycotics remains a controversial issue and should be studied in a controlled way. PMID- 2570526 TI - Association of genetic variation of the transforming growth factor-alpha gene with cleft lip and palate. AB - Complex segregation analysis of pedigrees having nonsyndromic cleft lip with or without cleft palate (CL/P) (Chung et al. 1986; Marazita et al. 1986) has shown that a major-locus model best explains the observed recurrence of CL/P in Caucasian families. To identify this major gene, we compared the frequencies of 12 RFLPs at five loci-epidermal growth factor, transforming growth factor-alpha, epidermal growth factor receptor, glucocorticoid receptor, and estrogen receptor in both a group of 80 subjects with nonsyndromic CL/P and 102 controls. These candidate genes were selected because studies in rodents had suggested their possible involvement in palatogenesis. A significant association was observed between two RFLPs at the transforming-growth-factor-alpha (TGFA) locus and the occurrence of clefting (P = .0047 and P = .0052). This suggests that either the TGFA gene itself or DNA sequences in an adjacent region contribute to the development of a portion of cases of CL/P in humans and provides an opportunity to begin to examine the molecular events underlying lip and palate formation. PMID- 2570527 TI - Hot spot of recombination within DXS164 in the Duchenne muscular dystrophy gene. AB - The DMD gene, which spans more than 2,000 kbp, has been assigned to band Xp21 of the X chromosome. Two subclones (PERT 87-1 and PERT 87-15) of the intragenic locus DXS164 physically are separated by approximately 60 kbp. Linkage studies were done in 49 informative DMD families by using the LINKAGE program. Crossing over between the loci studied occurred in four families. A recombination rate of 4% (support interval [Zmax-1] 1%-10%), which was 54 (support interval 14-135 fold) times higher than expected, was found with a maximum lod score of 13.50. These data suggest a hot spot for recombination within DXS164. PMID- 2570528 TI - Isodisomy of chromosome 7 in a patient with cystic fibrosis: could uniparental disomy be common in humans? AB - Maternal isodisomy for chromosome 7 was observed in a 4-year-old cystic fibrosis patient with very short stature. In an examination of 11 DNA polymorphisms spanning the entire length of chromosome 7, no paternal contribution could be shown in seven informative loci. Paternity was examined with probes for five polymorphic loci on the Y chromosome, for the pseudo beta-globin locus on chromosome 11 and by Jeffreys's hypervariable probes. The results with the latter gave a probability of 3.7 x 10(-9) for nonpaternity. Chromosomal examination revealed a centromeric heteromorphism of chromosome 7 in the mother, for which the proband was homozygous. Isodisomy of the patient was thus shown for the entire length of a maternal chromosome 7. The mechanisms leading to this isodisomy involve at least two events of abnormal cell division, events that may be meiotic, postzygotic, or both. This proband is the second reported maternal isodisomy; both were detected through homozygosity for CF. Both patients had short stature, which could have been caused by parental imprinting, since similar results have been observed in isodisomic mice. Homozygosity due to uniparental descent in man should be kept in mind as a mechanism for recessive disorders, especially for chromosome 7. PMID- 2570529 TI - Linkage heterogeneity between X-linked retinitis pigmentosa and a map of 10 RFLP loci. AB - In nine families in which X-linked retinitis pigmentosa (XLRP) is segregating, the lod scores of XLRP in a map of 10 RFLP loci were obtained by multipoint linkage analysis. The XLRP locus was located telomeric to DXS7 in seven of the families and centromeric to DXS7 in two of the families. Under the hypothesis of two XLRP loci, a heterogeneity (admixture) test was performed, providing significant evidence of heterogeneity in XLRP (P less than .01). No correlation was detected between the clinical manifestations of XLRP and the two different disease loci. PMID- 2570530 TI - Identification of two cosmids derived from within chromosomal band 3p21.1 that contain clusters of rare restriction sites and evolutionarily conserved sequences. AB - We have isolated large numbers of human recombinants from a cosmid library constructed from an interspecific (hamster/human) somatic cell hybrid whose only human component is an intact chromosome 3. Unique sequence probes were isolated from these recombinants and were used to localize them along the length of chromosome 3 by hybridization to a somatic cell hybrid deletion panel. We identified two cosmids, cA84 (D3S92) and cA199 (D3S93), derived from within chromosomal band 3p21.1. Both cosmids contained multiple rare restriction sites that were tightly clustered within the cosmids. We have therefore identified, in a region consistently deleted in a variety of lung cancers, two cosmids that may contain genes that are candidates for involvement in lung cancer. PMID- 2570532 TI - Pilus colonization factors of enterotoxigenic Escherichia coli isolated from infantile diarrheal patients in Papua New Guinea. AB - We examined the possibility of pilus colonization factor (or adhesin) production by 9 enterotoxigenic Escherichia coli isolated from infantile diarrheal patients in Papua New Guinea. By hydrophobicity, hemagglutination and bacterial agglutination with antisera against various known adhesins, none of the 9 strains produced known pilus adhesins. Three strains which produce heat-labile and heat stable enterotoxins may produce a new type of pili which is not hydrophobic and differs from common (type 1) pili. PMID- 2570531 TI - Roles of histamine, complement and xanthine oxidase in thermal injury of skin. AB - The pathogenesis of burn edema in the skin of rats appears to be related to a role for histamine, xanthine oxidase and oxygen radicals. Histamine and its metabolic derivatives increase the catalytic activity of xanthine oxidase (but not xanthine dehydrogenase) in rat plasma and in rat pulmonary artery endothelial cells. In thermally injured rats levels of plasma histamine and xanthine oxidase rise in parallel, in association with increases in uric acid. Burn edema is greatly attenuated by treatment of rats with the mast cell stabilizer, cromolyn, by complement depletion and by treatment with the H2 receptor antagonist, cimetidine, but is unaffected by neutrophil depletion. These studies suggest the following pathogenesis of burn edema: thermal trauma causes complement activation with anaphylatoxin release and mast cell secretion of histamine, leading to enhancement of xanthine oxidase activity and increased production of oxygen radicals which damage endothelial cells. PMID- 2570533 TI - Serologic evidence of hantaviral infections within small mammal communities of Baltimore, Maryland: spatial and temporal patterns and host range. AB - Serologic evidence was used to investigate the spatial and temporal distribution and host range of hantaviruses in small mammal communities in Baltimore, MD. Immunofluorescent antibody (IFA) reactive to a Baltimore rat isolate of Seoul virus was detected in 44% of 404 Norway rats captured at 4 park or residential locations; 21% of 121 meadow voles captured at 4 park locations possessed significant IFA titers to Prospect Hill virus. Evidence from plaque neutralization assay of rodent sera suggested that Seoul virus and Prospect Hill virus circulated concurrently in voles and rats, respectively, at 1 park. No cross infection of virus between these primary reservoir species was observed, as determined by higher specific neutralizing antibody titers to the characteristic virus for each host species. Only 4% of 449 house mice and 1% of 94 white-footed mice captured at the same sites as the primary host species were seropositive to hantaviruses, despite extensive demonstrated overlap in time and space with the primary host species. PMID- 2570534 TI - The effect of H2 antagonists on duration of action of suxamethonium in the parturient. AB - Fifteen patients who underwent elective Caesarean section were given sodium citrate, either alone or combined with oral cimetidine or ranitidine, as antacid premedication. No significant differences in time to 25 or 50% recovery of the first twitch of a train-of-four after administration of suxamethonium were found between any of the groups. It is concluded that H2-antagonists do not significantly affect the duration of action of suxamethonium in parturients, in contrast to the findings of a study in nonpregnant patients. PMID- 2570535 TI - [Fentanyl versus sufentanil basic anesthesia. Hypnotic effect, muscle rigidity and efficacy of competitive muscle relaxants]. AB - As induction agents for cardioanesthesia, sufentanil (S) and fentanyl (F) are usually employed in combination with nondepolarizing muscle relaxants. We investigated potential interactions of these opioids with the relaxant component, paying special regard to the role of muscular rigidity and opioid-induced alterations of hemodynamics. Narcotic anesthesia was induced randomly in 45 coronary artery bypass patients with either F (20 micrograms/kg) or S (4 micrograms/kg). After 6 min, neuromuscular blockade was initiated within each group randomly with either vecuronium (V) or pancuronium (P) (0.01 mg/kg each). During opiate administration, the times for cessation of spontaneous respiration and loss of responsiveness to verbal and tactile stimuli were measured. The degree of opiate-induced muscular rigidity, simultaneous changes in arterial paCO2, cardiac indices (CI) prior to opioid and relaxant administration, onset and recovery from neuromuscular blockade (by electromyographic train-of-four registration), and motor response to laryngoscopy and intubation were recorded. The onset of spontaneous apnea (TK = time to breathing upon command only) and unresponsiveness (TM = time to controlled mask ventilation) was significantly faster with S than with F. Muscular rigidity was moderate in 25% of patients and severe in 35%-40%, during the administration of both narcotics. No significant differences between S and F were observed. During ventilation by face mask, patients with clinically apparent rigidity showed a statistically significant mean increase in paCO2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570536 TI - Haemodynamic responses to acute blood loss: new roles for the heart, brain and endogenous opioids. AB - Information has come forward recently from several sources which provides new insights into the mechanisms that underlie the haemodynamic responses to acute blood loss. In unanaesthetised animals and human volunteers there are two distinct phases to these responses. At first, the engagement of baroreflexes results in a progressive rise in sympathetic vasoconstrictor drive and peripheral resistance, and the maintenance of arterial blood pressure at a near-normal level. When about one-third of blood volume has been lost, reflex sympathetic drive is switched off, and peripheral resistance and blood pressure fall abruptly to low levels despite a burst of vasopressin release. Research in conscious animals has now shown that the onset of this decompensatory phase is triggered by a signal from the heart, which activates an endogenous opioid mechanism in the brain. Activation of this mechanism can be prevented by administering a selective delta-receptor antagonist, or selective mu-receptor agonists (including alfentanil). It has not yet been established that this endogenous opioid mechanism is responsible for the decompensatory phase of acute blood loss in man, nor that it can be prevented or reversed by selective opioid agonists or antagonists. PMID- 2570537 TI - Prolonged neuromuscular blockade following the use of vecuronium for mediastinoscopy. PMID- 2570538 TI - Endocrine effects of early non-union of testis and epididymis and of cryptorchidism in the rat. AB - The aim of this study was to explore possible endocrine effects of early non union of testis and epididymis. In 16 days old Sprague-Dawley rats the testis and epididymis were separated to the level of the inferior epididymis artery (non union operation). Animals were killed at intervals from 30-58 days, and the plasma concentrations of total testosterone, total estrogens, LH, FSH and PRL were measured. The testes were studied by light microscopy. Groups of rats made cryptorchid and sham-operated rats were used as controls. Although their testes were scrotal, the non-union operated animals had testosterone and estrogen concentrations similar of cryptorchid animals, and significantly (p less than 0.05) lower than sham-operated animals. The LH- and FSH-concentrations were significantly (p less than 0.05) elevated suggesting a primary lesion in both Leydig- and Sertoli-cells. Towards puberty FSH increased in the non-union operated animals, while FSH values declined in the cryptorchid and sham-operated animals. These FSH-patterns probably reflect the existence of different pathogenic mechanisms in the non-union operated rats and the rats with cryptorchid testes. PMID- 2570539 TI - Narcotic analgesics and debrisoquine polymorphism. PMID- 2570541 TI - Role of brain microvessels and choroid plexus in cerebral metabolism of leukotrienes. PMID- 2570540 TI - Incidence of anaphylactic self-treatment in an outpatient population. AB - Insect-allergic and food-allergic patients with a history of previous anaphylactic reactions were retrospectively and prospectively evaluated for incidence of self-administration of epinephrine or antihistamines. Participants were prescribed either Ana-KitR or Epi-PenR dosage forms. In 33 patients, 82 incidents were noted during 1,252 patient-months, an average of 0.78 incidents per patient-year (PPY). A mean of 0.38 epinephrine uses PPY was recorded, ranging from 0 to 2.82 doses PPY. Antihistamine uses averaged 0.69 PPY, ranging from 0 to 4.47 doses PPY. Antihistamines alone quelled reactions in 13 of 17 incidents involving Ana-Kits. Food-allergic patients were more likely to self-medicate, with 1.93 anti-allergic drug uses PPY, compared with 0.49 uses PPY for insect allergic patients. Six of 22 insect-related epinephrine uses occurred during winter months. No statistically significant difference was noted between drug administration rates determined retrospectively and prospectively. PMID- 2570542 TI - The role of arachidonic acid and its metabolites in the release of neuropeptides. PMID- 2570543 TI - Modulation of arachidonate turnover in cerebral phospholipids. AB - The effects of the neurotransmitters NE and 5HT on the turnover of AA in cerebral PL were investigated in slices of rat brain cortex. Incorporation of 3[H]-AA into individual PL was first analyzed in accordance with a closed two-compartmental model. Apparent rates of deacylation and reacylation as well as sizes of the metabolically active PL-bound AA pools were calculated. It was found that rates of reacylation of individual PL varied markedly, while deacylation rates remained within a relatively narrow range. The rate of PI acylation was found to be the most rapid, while the rate of PS acylation was the slowest observed. The pool of PL-bound AA that is readily accessible to deacylation-reacylation processes was distributed differentially among the various PL, with more than 50% of this pool in PI; but only 0.75% of the PI content was associated with this pool. Both NE and 5HT enhanced the incorporation of 3[H]-AA into PI in a dose-related manner, while they attenuated its incorporation into other PL. Pharmacological studies indicated that the neurotransmitter effects were not mediated by known NE or 5HT receptors and that a functional presynaptic reuptake system was not required for these effects. The observed effects did not appear to be related to the formation of hydrogen peroxide by the action of MAO on the neurotransmitter. Examination of the structure-activity relationships indicated that the presence of two hydroxyl groups in the aromatic ring was needed for attenuating 3[H]-AA incorporation into PC, whereas an active catechol nucleus with an additional hydroxyl group in the beta position of the side chain appeared to enhance 3[H]-AA incorporation into PI. Results obtained with the phospholipase A-2 inhibitor mepacrine and the acyltransferase inhibitor THC suggest that NE attenuates PL acylation by activating phospholipase A-2, but it concomitantly enhances PI acylation by selectively stimulating a PI-specific arachidonyl transferase via mechanisms that have not yet been elucidated. PMID- 2570544 TI - Arachidonic acid metabolites in the rat and human brain. New findings on the metabolism of prostaglandin D2 and lipoxygenase products. PMID- 2570546 TI - Pain management in sickle cell disease. Rationale and techniques. PMID- 2570545 TI - Neural systems involved in fear-potentiated startle. AB - The acoustic startle reflex offers a number of advantages for analyzing the neural bases of behavior. Startle apparently is mediated by excitatory amino acids at several of the synapses that comprise the short-latency startle pathway. The reflex is modulated by a variety of neurotransmitters at both the spinal and the supraspinal levels. In addition to showing habituation and sensitization, startle is increased in the presence of a fear stimulus. This may result from activation of the central nucleus of the amygdala which projects directly tot he acoustic startle pathway. A major challenge for future studies will be to determine what neurotransmitters, which are known to modulate startle, are involved in habituation, sensitization, and fear conditioning and to begin to analyze these processes at a cellular level. PMID- 2570547 TI - Catecholamine depletion produces irrepressible saccadic eye movements in normal humans. AB - To determine the effect of catecholamine depletion on ocular motor pathways in humans, we studied the eye movements of 3 normal subjects who received the drug metyrosine (alpha-methylparatyrosine). This drug temporarily depleted dopamine and norepinephrine, as measured by a reduction in the metabolite, 3-methoxy-4 hydroxy-phenylethyleneglycol (MHPG). Saccadic, pursuit, and vestibulo-ocular eye movements were recorded using infrared oculography with subjects both on placebo and on metyrosine. The most consistent effect observed with metyrosine was an increase in the amplitude and frequency of saccadic intrusions during fixation and pursuit. Two of the 3 subjects also had shortened time constants for the vestibulo-ocular reflex, attributable in part to the sedative effect of catecholamine depletion. The increase in saccadic intrusions implies that catecholamines modulate the activity of a subpopulation of suppressor motor neurons in the human brainstem. PMID- 2570548 TI - The biochemistry of P-glycoprotein-mediated multidrug resistance. PMID- 2570549 TI - Genetic contributions to the variation among rabbits of liver microsomal deoxycorticosterone synthesis. AB - Outbred New Zealand white rabbits exhibit two phenotypes, 21H and 21L, corresponding to rates greater than or less than 1 nmol/min/mg, respectively, for liver microsomal progesterone 21-hydroxylase activity. In contrast, the inbred strain III/J exhibits only the 21L phenotype. Two 21H male New Zealand white rabbits were mated with several female III/J rabbits to produce a total 46 progeny. Both the 21H and 21L phenotypes were evident among male and female offspring in roughly equal numbers. Backcrosses between 21L progeny and III/J rabbits exhibit only the 21L phenotype, whereas 21H offspring yield both 21H and 21L progeny when backcrossed to the 21L inbred strain III/J. These results are consistent with autosomal dominant inheritance of the 21H phenotype. Analysis of Southern blots of genomic DNA digested with the restriction endonuclease KpnI reveals 20-, 13-, and 9-kb fragments that hybridize with a probe derived from the 3'-untranslated region of the 21-hydroxylase cDNA. The 13-kb band is not observed for strain III/J or 21L progeny of strain III/J crossed with 21H rabbits, but it is detected for both 21H fathers and 21H progeny indicating that the genetically determined difference of 21-hydroxylase expression is inherited cis to the gene for P450IIC5, the hepatic progesterone 21-hydroxylase. Electrophoretic analysis of P450IIC5 synthesized in vitro from mRNA isolated from 21L and 21H rabbits reveals that little or no P450IIC5 is synthesized from 21L mRNAs. A second immunoreactive, electrophoretically distinct protein is synthesized from both 21L and 21H mRNAs to a similar extent but in lesser amounts than P450IIC5. The second protein could represent either an allozymic form of the enzyme or the product of a distinct locus. Thus, it is likely that distinct structural genes for P450IIC5 contribute to the differences in P450-mediated metabolism in 21L as compared to 21H rabbits. PMID- 2570550 TI - HLA types in patients with rheumatoid arthritis developing leucopenia after both gold and sulphasalazine treatment. AB - HLA types, especially HLA-DR3, are associated with the development of toxic reactions in patients with rheumatoid arthritis after treatment with gold or D penicillamine. In this study, after treatment with sulphasalazine, leucopenia was observed in three patients, who all had a history of leucopenia after previous gold treatment. The HLA types of these patients did not include HLA-DR3; the two patients developing mild leucopenia had HLA-DR2 and the one developing agranulocytosis had HLA-DR4. PMID- 2570551 TI - Combined revascularization of coronary and femoral arteries: a proposed alternative. AB - Two patients with both coronary artery disease and leg ischemia were successfully treated with a combined revascularization procedure. Coronary arteries were bypassed with in situ internal mammary artery grafts, and bilateral femoral arteries were bypassed with expanded polytetrafluoroethylene grafts descended from the ascending aorta through the preperitoneal abdominal wall tunnel. Both patients recovered well and experienced no angina or claudication. PMID- 2570552 TI - Interaction between R 56865 and alpha-adrenoceptors in the pithed rat. AB - In pithed normotensive rats, the benzothiazolamine derivative R 56865 in high doses exhibits a competitive antagonism of alpha 1-adrenoceptor-mediated vasoconstrictions. The absence of a depression of the maximum of the dose response curve of ST 587 and the very moderate attenuation of the maximal B-HT 920-induced increase in diastolic blood pressure (BP) confirms the lack of major calcium entry blocking properties of R 56865 for alpha-adrenoceptor-activated calcium channels in vitro. In doses up to 10(-5) mol/kg, the interaction of R 56865 with the sympathetic neurotransmission can solely be explained by alpha 1 adrenoceptor blockade. This was confirmed by the comparable antagonism of the selective alpha 1-adrenoceptor antagonist prazosin in a concentration of 6 x 10( 8) mol/kg. In contrast to the isolated rat aorta, where R 56865 showed an allosteric interaction with the NA binding site on the alpha 1-adrenoceptor, R 56865 acts like an alpha 1-adrenoceptor antagonist of the competitive type in vivo. PMID- 2570553 TI - Cysteamine and pantethine effects on passive avoidance behavior, shuttle box learning, open-field activity, striatal catecholamines and somatostatin. AB - The effects of cysteamine and pantethine were compared on different behavioral tests and neurochemical parameters in rats. Cysteamine, administered in high dose (3.90 mM/kg s.c.), decreased the locomotor and rearing activities of rats, while it slightly but not significantly increased the avoidance latency in a passive avoidance test. Pantethine, 24 hr after its administration, significantly increased the dihydroxyphenyl acetic acid (DOPAC) levels in the striatum. Cysteamine slightly reduced the DOPAC level without influencing the catecholamine levels in this brain area. The striatal somatostatin concentration was reduced 24 hr after the administration of cysteamine, while pantethine did not influence it. After repeated daily injections of pantethine, the drug facilitated the shuttle box learning process and increased the intertrial and open-field activities of the animals. Cysteamine only slightly increased the locomotion and rearing and did not influence the shuttle box learning. Both pantethine and cysteamine slowed the rate of the "body weight increase" of the animals when compared to a saline treated group. These findings suggest that the locomotor activation induced by pantethine 24 hr after its administration plays an important role in its behavioral effects. It might be that the striatal dopaminergic transmission, modified by administration of pantethine, plays some role in the higher locomotor activity induced by the substance. PMID- 2570554 TI - Alpha-adrenoceptor antagonistic effects and antihypertensive activity of SGB-1534 in conscious normotensive and deoxycorticosterone acetate (DOCA)-salt hypertensive dogs: comparison with prazosin. AB - In conscious normotensive and deoxycorticosterone acetate (DOCA)-salt hypertensive dogs, alpha-adrenoceptor antagonistic and antihypertensive actions of SGB-1534, given orally, were compared with those of prazosin. SGB-1534 and prazosin did not markedly influence systemic blood pressure (SBP) and heart rate (HR) in normotensive dogs. These alpha 1-adrenoceptor antagonists inhibited SBP and HR increases induced by i.v. adrenaline in a dose-dependent manner. In DOCA salt hypertensive dogs, persistent hypotension, accompanied by a reflex tachycardia, occurred after oral administration of SGB-1534 and prazosin. The hypotensive activity of SGB-1534 was much more potent in magnitude and duration than that of prazosin. In isolated canine mesenteric and femoral arteries, SGB 1534, compared to prazosin, elicited a more potent inhibition on the contractile response to a relatively selective alpha 1-adrenoceptor agonist, phenylephrine. It appears that the antihypertensive activity of SGB-1534, like prazosin, is nearly in parallel with its alpha 1-adrenoceptor antagonistic effect. PMID- 2570555 TI - Electrophysiological evaluation of the beta-blocking properties and direct membrane effects of l-moprolol and its enantiomer d-moprolol. AB - The electrophysiological and beta-blocking properties of l- and d-moprolol have been studied in sheep cardiac Purkinje fibers. l-Moprolol (0.3-100 nM) dose dependently antagonized the inotropic (ED50 = 1.2 +/- 0.4 nM) and chronotropic (ED50 = 5.8 +/- 0.6 nM) effects of isoprenaline. d-Moprolol (0.3-100 nM) was fully devoid of beta-blocking properties under the same experimental conditions. Both isomers had the same electrophysiological profile: they did not influence action potential characteristics, contractility and normal automaticity up to 1 microM. On the whole, l-moprolol appears to be a potent beta-blocker, exerting membrane depressant effects only at a concentration 2000 times higher than those exerting effective beta-blockade. PMID- 2570556 TI - Effects of the anti-dementia drug hopantenate calcium upon striatal dopaminergic neurons in young and aged rats. AB - In the present study we investigated the effects of the anti-dementia drug calcium D-(+)-4-(2,4-dihydroxy-3,3-dimethyl-butyramido) butyrate hemihydrate (hopantenate) on the dopaminergic neurons of rats, and also compared the effects of the drug on dopaminergic neurons in young adult rats (4 months old) and aged rats (21 months old). Hopantenate 1000 mg/kg, p.o. significantly increased striatal dopamine (DA) levels, but displayed almost no effect upon the DOPAC and HVA levels. Furthermore, we investigated the effects of hopantenate upon tyrosine hydroxylase activity by examining NSD-1015-induced L-DOPA accumulation and found that hopantenate 1000 mg/kg, p.o. significantly increased the L-DOPA accumulation. In addition, comparing the effect of hopantenate on dopaminergic neurons in young adult rats and aged rats, we found that the striatal DA, DOPAC and HVA levels were decreased as a concomitant of aging, and hopantenate 1000 mg/kg, p.o. significantly increased DA and DOPAC levels in both ages. The above results clearly indicate that hopantenate enhanced DA biosynthesis by stimulating the activity of tyrosine hydroxylase. Furthermore, the results of hopantenate upon dopaminergic neurons in young adult rats and aged rats suggest that sensitivity to the drug may not be different with age, though the striatal DA, DOPAC and HVA levels of rats were decreased as a concomitant of aging. PMID- 2570557 TI - Ammonium assimilation in Proteus vulgaris, Bacillus pasteurii, and Sporosarcina ureae. AB - No active uptake of ammonium was detected in Proteus vulgaris, Bacillus pasteurii, and Sporosarcina ureae, which indicates that these bacteria depend on the passive diffusion of ammonia across the cell membrane. In P. vulgaris the glutamine synthetase-glutamate synthase (GS-GOGAT) pathway and glutamate dehydrogenase (GDH) were present, and these enzymes exhibited high affinities for ammonium. In B. pasteurii and S. ureae, however, no GS activity was detected, and GOGAT activity was only present in S. ureae. GDH enzymes were present in these two organisms, but showed only low affinity for ammonium, with apparent Km-values of 55.2 mM in B. pasteurii and 36.7 mM in S. ureae, respectively. These observations explain why P. vulgaris is able to grow at neutral pH and low ammonium concentration (2 mM), while B. pasteurii and S. ureae require high ammonium concentration (40 mM) and alkaline pH for growth. PMID- 2570558 TI - [The fermentation diagnosis and histologic studies in blood and the liver of surviving rats after 1 and 2 administrations of a median toxic dose of parathion methyl. 1. Results of studies on the activities of the plasma enzymes AlAT, AsAT, AP, and gamma-GT]. AB - Clearly pronounced changes in plasma enzyme activities were observed 24 hours after one and two applications of parathion methyl (PM). The most strongly pronounced rise was in activity of aspartate-amino transferase and was recorded from both sexes. PM application caused sizeable changes in male rats. Comparison between reactions after one and two applications revealed more intensive changes in males after the 1st application. Somewhat larger differences were recordable in some cases between the 1st and 2nd applications. Closer agreement between reaction intensities after the 1st and 2nd PM applications was recordable from female rats. In either sex in response to one single and repeated application parameters returned to normal within nearly one and the same period of time. No secured relationships between activities prior to and after application of PM could be derived from the assays of gamma-glutamyl transpeptidase (gamma-GT). Both increase and decrease of determined enzyme activities were actually recorded. Activities were highly variable in the control groups. Extreme rises were recordable in some cases from both control and experimental groups. No conclusions so far can be drawn as to "normal values" in rat plasma, possible sex related differences, and behaviour of enzyme activity in response to one single or repetitive applications. Hence, within common investigations of acute toxicity of alkylphosphates, gamma-GT was found to be not suitable for detection of damage to the liver or other organs. PMID- 2570559 TI - [The fermentation diagnosis and histologic studies in blood and the liver of surviving rats after 1 and 2 administrations of a median toxic dose of parathion methyl. 2. Tyrosine aminotransferase and pathologico-morphological findings in the liver]. AB - Tyrosine-amino transferase, an enzyme induced to the liver via glucocorticoids, caused clearly measurable activity rises in either sex, 24 hours after 1 and 2 applications. Increased levels continued to recordable from males on the 3rd day from application, whereas enzyme activities in females had already returned to normal by that time. Histological examination of liver sections, following 1 and 2 applications of parathion methyl (PM), revealed the tissue pattern of clearly manifest toxic hepatosis with detectable mesenchymal reactions. Histologically detectable damage reached its maximum on the 2nd and 3rd days from application. Sex-related differences were not recordable. Damage in response to the 2nd application was characterised by faster mesenchymal activation and higher frequency of cell necroses. The morphological picture of the liver was rated as having returned to normal, 14 days after 1 and 2 applications of PM. These results should not be interpreted as evidence to the involvement of specific, non cholinergic mechanisms in the processes of damage observed. The alterations might rather be attributed to unspecific disorders in the energy balance or to the effect of "stress" during intoxication. PMID- 2570560 TI - Papillary adenoma of the stomach. Pathologic and immunohistochemical study. AB - A total of 13 gastric papillary adenomas composed of 8 papillary and 5 papillotubular adenomas were examined pathologically and immunohistochemically. They showed a dome-like or pedunculated appearance and were located at the antrum, except for one adenoma. Histologically, the adenoma cells showed atypia in varying degree and focal adenocarcinoma was noted in seven lesions. The number of goblet cells was apparently smaller in the papillary than in the tubular portion. Lysozyme was present at the supranuclear region in most papillary adenoma cells, whereas it was concentrated in Paneth's granules in tubular adenoma cells. No difference was found in the distribution and frequency of carcinoembryonic antigen, secretory component, and carbohydrate antigen CA 19-9 between papillary and tubular adenomas. Paucity of endocrine cells also characterized gastric papillary adenoma. Different phenotypic expressions might reflect the difference in histogenesis between papillary adenoma and tubular adenoma. PMID- 2570561 TI - Incidence of preneoplastic foci in rat liver dependent on different promoting schemes. PMID- 2570562 TI - Detection of HIV-1 DNA in different subsets of human peripheral blood mononuclear cells using the polymerase chain reaction. Rapid communication. AB - The presence of HIV-1 proviral DNA was determined in CD4 cells, CD8 cells and macrophages/monocytes obtained from peripheral blood of 8 HIV-1 infected persons. Using the polymerase chain reaction (PCR) we were able to detect proviral DNA in the extracts of only 10(2)-10(3) CD4 (T4) cells. In contrast, 10(5) CD8 cells did not contain detectable amounts of proviral DNA. Surprisingly, in four of our eight patients studied no HIV-1 DNA was found in macrophages. In peripheral blood, every hundred T4 cell may be infected with HIV-1. PMID- 2570563 TI - [Stomach cancer with tumor endocrine cells in the presence of chronic ulcer]. AB - A case of stomach carcinoma was examined in a 45-year-old male by using histochemical and electron microscopic techniques. The carcinoma had arisen from chronic gastric ulcer accompanied with fundic gland hyperplasia and located in the body of the stomach. Histologically, it was a nondifferentiated scirrhous carcinoma involving signet ring and minor polymorphic cells with eosinophilic cytoplasm. Electron microscopically, the tumors had cells containing a large body of mucoidal granules, great quantities of cells having various endocrine granules, and cells showing mixed endocrine-exocrine secretion. It was suggested that there might be a pathogenetic relationship of fundal cell hyperplasia and signet ring cell carcinoma development, on the one hand, and the existence of tumor endocrine cells, on the other. PMID- 2570564 TI - The use of seclusion: a comparison of two psychiatric intensive care units. AB - The use of seclusion within a psychiatric intensive care unit in a South Australian metropolitan mental hospital was documented over a ten week period. The seclusion rate within the unit was 32% of all admissions and 34% of new admissions. The overall seclusion rate for the hospital was 5.4% of all admissions and 6.3% of new admissions, somewhat higher than in the United Kingdom but considerably lower than in the Eastern United States. A comparison was then made between consecutive new admissions (30 secluded and 30 non-secluded) to this unit and to a similar unit without a seclusion room in the other mental hospital in the State. Although seclusion offered no clear advantages in terms of duration of admission, levels of medication or relapse rates, it appeared to reduce the level of dangerousness in the unit, thereby enhancing staff morale. The overall mean daily total of neuroleptic medication was about 1,200 mg chlorpromazine equivalent, somewhat less than in comparable units in the United States and Europe. PMID- 2570565 TI - The inflight use of three pharmacological preparations: Humibid, Nicorette, and Seldane. PMID- 2570566 TI - Chromosomal location of the gene encoding the murine acute-phase protein serum amyloid P-component (SAP). AB - A full-length cDNA clone, pmSAP3, encoding, the serum P component (SAP), has been used to search for DNA fragment length variation among mouse strains previously analyzed for differences in endogenous SAP levels. Three alleles were found using EcoRI-digested DNA. The finding of a single 5.4-kb fragment, allele d, in DNA from DBA/2J mice suggests the presence of a single Sap locus. Segregation of DNA fragment associated with Sapb and Sapd alleles was analyzed in three sets of recombinant inbred (RI) strains. The strain distribution pattern found for the Sap alleles was identical to that of alleles of Ly-9 in 43 individual RI strains, suggesting tight linkage with Ly-9 on mouse chromosome 1. In the BXD RI strains, the SDP of the Sap locus, defined by the difference in the endogenous SAP level, is also identical to the SDP of the DNA fragments. We propose to redesignate the Sap locus to include both the structural element defined by the DNA polymorphism and the regulatory element involved in the regulation of SAP synthesis. The Sap locus is the major genetic element contributing to the regulation of SAP production. Other genetic factors are also involved, as shown by the presence of nonparental phenotypes in the individual BXH RI strains. PMID- 2570568 TI - Spontaneous excretion of D-alanine in urine in mutant mice lacking D-amino-acid oxidase. AB - Urine from mutant mice lacking D-amino-acid oxidase contained a large amount of alanine compared with that from normal mice. Urinary alanine of the mutant mice was sensitive to D-amino-acid oxidase. H.p.l.c. showed that about 94% of the urinary alanine had the D-configuration. These results suggest that D-amino-acid oxidase functions to decompose D-amino acid(s) in normal mice. PMID- 2570567 TI - Multiple actions of beta-adrenergic agonists on skeletal muscle and adipose tissue. PMID- 2570569 TI - The beta-adrenergic radioligand [3H]CGP-12177, generally classified as an antagonist, is a thermogenic agonist in brown adipose tissue. AB - The effect of CGP-12177, originally developed as a radioligand with antagonist properties for binding studies of beta-adrenergic receptors, was investigated in brown adipose tissue. Contrary to expectations, CGP-12177 showed clear agonist properties in experiments with hamster brown-fat cells, with a maximal effect in stimulating oxygen consumption similar to that of the physiological stimulator noradrenaline, and also with a potency similar to that of noradrenaline [EC50 (50% effective concn.) approx. 70 nM]. This value could be contrasted with the very high affinity of CGP-12177 (KD about 1 nM) for ligand-binding sites on the cells. It is therefore suggested that the high-affinity binding site may not be the one that mediates the CGP-12177-stimulated thermogenesis in isolated cells. Also, when injected into cold-adapted rats, CGP-12177 stimulated non-shivering thermogenesis similarly to noradrenaline. This observation, in conjunction with the reported low general sympathomimetic effect of CGP-12177, may indicate that CGP-12177 could be of interest for the development of anti-obesity drugs. PMID- 2570570 TI - Glutamine-dependent carbamoyl-phosphate synthetase and other enzyme activities related to the pyrimidine pathway in spleen of Squalus acanthias (spiny dogfish). AB - The first two steps of urea synthesis in liver of marine elasmobranchs involve formation of glutamine from ammonia and of carbamoyl phosphate from glutamine, catalysed by glutamine synthetase and carbamoyl-phosphate synthetase, respectively [Anderson & Casey (1984) J. Biol. Chem. 259, 456-462]; both of these enzymes are localized exclusively in the mitochondrial matrix. The objective of this study was to establish the enzymology of carbamoyl phosphate formation and utilization for pyrimidine nucleotide biosynthesis in Squalus acanthias (spiny dogfish), a representative elasmobranch. Aspartate carbamoyltransferase could not be detected in liver of dogfish. Spleen extracts, however, had glutamine dependent carbamoyl-phosphate synthetase, aspartate carbamoyltransferase, dihydro orotase, and glutamine synthetase activities, all localized in the cytosol; dihydro-orotate dehydrogenase, orotate phosphoribosyltransferase, and orotidine 5'-decarboxylase activities were also present. Except for glutamine synthetase, the levels of all activities were very low. The carbamoyl-phosphate synthetase activity is inhibited by UTP and is activated by 5-phosphoribosyl 1 pyrophosphate. The first three enzyme activities of the pyrimidine pathway were eluted in distinctly different positions during gel filtration chromatography under a number of different conditions; although complete proteolysis of inter domain regions of a multifunctional complex during extraction cannot be excluded, the evidence suggests that in dogfish, in contrast to mammalian species, these three enzymes of the pyrimidine pathway exist as individual polypeptide chains. These results: (1) establish that dogfish express two different glutamine dependent carbamoyl-phosphate synthetase activities, (2) confirm the report [Smith, Ritter & Campbell (1987) J. Biol. Chem. 262, 198-202] that dogfish express two different glutamine synthetases, and (3) provide indirect evidence that glutamine may not be available in liver for biosynthetic reactions other than urea formation. PMID- 2570571 TI - Sex-related difference in the inductions by perfluoro-octanoic acid of peroxisomal beta-oxidation, microsomal 1-acylglycerophosphocholine acyltransferase and cytosolic long-chain acyl-CoA hydrolase in rat liver. AB - Inductions by perfluoro-octanoic acid (PFOA) of hepatomegaly, peroxisomal beta oxidation, microsomal 1-acylglycerophosphocholine acyltransferase and cytosolic long-chain acyl-CoA hydrolase were compared in liver between male and female rats. Marked inductions of these four parameters were seen concurrently in liver of male rats, whereas the inductions in liver of female rats were far less pronounced. The sex-related difference in the response of rat liver to PFOA was much more marked than that seen with p-chlorophenoxyisobutyric acid (clofibric acid) or 2,2'-(decamethylenedithio)diethanol (tiadenol). Hormonal manipulations revealed that this sex-related difference in the inductions is strongly dependent on sex hormones, namely that testosterone is necessary for the inductions, whereas oestradiol prevented the inductions by PFOA. PMID- 2570574 TI - Urodilatin and beta-ANF: binding properties and activation of particulate guanylate cyclase. AB - Urodilatin (ANF-(95-126] and beta-ANF, the antiparallel dimer of ANF-(99-126), are naturally occurring members of the ANF family. We studied their receptor binding properties in human platelets and Triton-solubilized membranes from bovine adrenal cortex and their ability to activate particulate guanylate cyclase in bovine adrenal cortex. In human platelets containing R2-receptors not coupled to particulate guanylate cyclase urodilatin binds with similar affinity as ANF (99-126) (KD: 55 pM), whereas beta-ANF has an affinity lower than the truncated ANF-(103-123) (KD: 295 pM and 154 pM). Scatchard analysis indicates one binding site for urodilatin as well as for beta-ANF. In adrenal cortex containing predominantly R1-receptors coupled to particulate guanylate cyclase, urodilatin binds with a higher affinity (KD: 30 pM) than ANF-(99-126) (KD: 52 pM) and stimulates to a similar extent to ANF-(99-126) (about two fold at 1 muM), whereas beta-ANF has a smaller affinity (KD: 120 pM) and stimulates particulate guanylate cyclase to a lower extent than ANF-(99-126). The data from platelets and adrenal cortex show that beta-ANF has low binding affinities but stimulates particulate guanylate cyclase, whereas urodilatin appears to be a physiological R1-agonist. PMID- 2570573 TI - Age-related changes in linoleate and alpha-linolenate desaturation by rat liver microsomes. AB - The first and rate limiting step in the conversion of linoleic and alpha linolenic acid is catalyzed by the delta - 6 - desaturase (D6D) enzyme. Rat liver microsomal D6D activity decreases on linolenic acid at a rate proportional to the animal age; on alpha-linolenic acid the decrease in D6D activity begins only later than on linoleic acid. The fatty acid composition of liver microsomes determined by gas chromatographic analysis confirms the impairment of the enzymatic activity directly measured. Our data indicate a correlation between aging and D6D activity impairment. The loss of D6D activity may be a key factor in aging through altering the eicosanoid balance. PMID- 2570572 TI - Alpha 1- and beta-adrenergic regulation of intracellular Ca2+ levels in brown adipocytes. AB - In order to monitor changes in cytosolic Ca2+ levels, brown-fat cells were incubated with the fluorescent Ca2+-indicator fura-2 and the fluorescence intensity ratio followed. The addition of norepinephrine led to a rapid and persistent increase in the cytosolic Ca2+ level, which was dose-dependent with a maximal effect at about 1 microM. The response was diminished in the absence of extracellular Ca2+ and was inhibited more efficiently by phentolamine and prazosin than by propranolol or yohimbine, indicating alpha 1-adrenergic mediation. Accordingly, selective alpha 1-adrenergic stimulation also increased the cytosolic Ca2+ level. However, selective beta-adrenergic stimulation, as well as the adenylate cyclase activator forskolin, were also able to increase the cytosolic Ca2+ level in these cells to a certain extent. It was concluded that the major part of the increase in cytosolic Ca2+ was mediated, as in other cell types, via alpha 1-adrenergic receptors, but that Ca2+ levels were also positively modulated by a cAMP-mediated process. These observations are discussed in relation to known alpha 1/beta synergisms in brown adipose tissue. PMID- 2570575 TI - Transient shut off of Escherichia coli heat shock protein synthesis upon temperature shift down. AB - A moderate downward shift in growth temperature (37 to 30 degrees C in strain B/r and 37 to 24 degrees C in strain K-12) was found to depress markedly the synthesis of major heat shock proteins GroEL and DnaK in E. coli. The depression was transient and cancelled gradually to a new steady state level, taking 60-80 min. The synthesis of beta-galactosidase directed by transcription initiated at the groE promoter behaved similarly, suggesting that this regulation, termed "reverse heat shock response", occurs at the transcriptional level. PMID- 2570576 TI - The inhibition of cytoplasmic acetoacetyl-CoA thiolase by a triyne carbonate (L 660, 631). AB - The compound L-660, 631 (2-oxo-5-(1-hydroxy-2,4,6-heptatriynyl)-1,3-dioxolane-4 heptanoic acid), a natural product isolated from an Actinomycete culture, was found to inhibit rat liver cytosolic acetoacetyl-CoA thiolase, the first step in the cholesterol biosynthesis pathway, with an IC50 of 1.0 x 10(-8) M. The inhibitor had no effect on other sulfhydryl containing enzymes of lipid synthesis such as HMG-CoA synthase, HMG-CoA reductase, and fatty acid synthase. When tested in cultured human liver Hep G2 cells the compound inhibited the incorporation of 14C-acetate and 14C-octanoate into sterols 56% and 48% respectively at 3 x 10(-6) M with no effect on fatty acid synthesis. No noticeable effect was seen on fatty acid biosynthesis. This strongly suggests that the locus of inhibition of acetate incorporation into sterols found with this compound is the acetoacetyl-CoA thiolase step in the cholesterol biosynthesis pathway. PMID- 2570577 TI - Effect of cellular Ca2+ loading on alpha 1-agonist or protein kinase C activators mediated stimulation of phosphorylase "a" in liver cells. AB - Long chain unsaturated fatty acids stimulate phosphorylase "a" activity in liver cells. Similar degree of activation was achieved by increasing cellular Ca2+ content or by treatment with agents other than oleate, like 1,2-diolein or phorbol esters, sharing in common their ability to activate protein kinase C. In Ca2+-loaded liver cells only phenylephrine was capable of inducing a further stimulation of phosphorylase "a" activity. It is concluded that: 1) The state of activation of protein kinase C may play a role in the hormonal control of liver glycogen metabolism; 2) alpha 1-agonist-mediated activation of phosphorylase "a" can occur by a mechanism which is not related to a Ca2+-dependent activation of protein kinase C. PMID- 2570580 TI - Combination methotrexate and sulfasalazine in the management of rheumatoid arthritis: case observations. AB - Four patients with rheumatoid arthritis received a combination of methotrexate and sulfasalazine for a mean of 24 months (range 20-28 months). All 4 patients experienced clinical improvement, with a reduction in the number of involved joints and in morning stiffness. In all 3 patients who had previously taken methotrexate, we were able to reduce the dosage, and the prednisone dosage was reduced in 2 of 3 patients who had previously taken that drug. No serious toxicity was observed in any patient. PMID- 2570578 TI - Effect of heat-stable and heat-labile enterotoxins of Escherichia coli on intestinal brush border membrane enzymes of mice. AB - The activities of intestinal brush border membrane (BBM) enzymes alkaline phosphatase, maltase, lactase, sucrase, gamma-glutamyl transpeptidase and leucine aminopeptidase were determined in intestinal homogenates and purified BBMs from control, heat-stable and heat-labile enterotoxin treated mice. The activities of all the enzymes except lactase were decreased significantly (p less than 0.01) in homogenates while increased significantly (p less than 0.001) in BBMs of experimental groups as compared to controls. Calmodulin activities were increased significantly (p less than 0.01) as compared to control in heat-stable enterotoxin treated mice but remained unaltered in heat-labile enterotoxin treated mice. DNA contents of intestinal homogenates were decreased in experimental groups demonstrating the decrease in cell number in these groups. The altered BBM enzyme activities could not be attributed to changes in calmodulin activities. The increase in enzyme activities in BBMs may reflect a compensatory phenomenon in the remaining cells. PMID- 2570579 TI - The changes in activities of some ammonia metabolizing enzymes in liver and brain of rats intoxicated by chronic administration of acetaldehyde. PMID- 2570581 TI - Selective blockade of alpha 1-adrenoceptors of porcine vascular smooth muscle cells by bunazosin. AB - The characteristics of bunazosin binding to alpha-adrenoceptors in the porcine aortic membranes were investigated using [3H]prazosin and [3H]yohimbine binding assays to identify alpha 1- and alpha 2-adrenoceptors, respectively. The extent of the inhibition (Ki values) of [3H]prazosin binding to alpha 1-adrenoceptors induced by bunazosin was 0.29 nmol/l and about the same as that induced by prazosin (Ki = 0.10 nmol/l). The Ki value of bunazosin inhibition of [3H]yohimbine binding to alpha 2-adrenoceptors was 350 nmol/l. There was over a 1000-fold difference in Ki value for bunazosin between alpha 1-and alpha 2 adrenoceptors. Thus, bunazosin is a highly alpha 1 selective agent in vascular smooth muscle. PMID- 2570582 TI - [The effect of a combination of ridazolol and molsidomine on hemodynamics and left ventricular function in dilated cardiomyopathy]. AB - Haemodynamics and Left Ventricular Function in Patients with Dilated Cardiomyopathy after a Combination of Ridazolol and Molsidomine. To evaluate the haemodynamic properties of the new beta-blocker ridazolol alone and in combination with the vasodilator molsidomine a randomized study was carried out in 12 patients with dilated cardiomyopathy (NYHA II/III). beta-Blockade leads to a decreasing tendency in blood pressure and heart rate. Pulmonary capillary wedge pressure (PC) was not significantly altered and ejection fraction remained unchanged. Molsidomine showed a decrease of PC. The combination of both drugs revealed both the positive haemodynamic effects of molsidomine and the reduced heart rate of beta-blockade. Negative side effects could not be observed. PMID- 2570583 TI - Planning for the future of occupational health nursing. Part II: Comprehensive membership survey. PMID- 2570584 TI - Astrocytic swelling in traumatic-hypoxic brain injury. Beneficial effects of an inhibitor of anion exchange transport and glutamate uptake in glial cells. AB - Swelling of brain slices is shown to occur in response to elevated potassium levels or glutamate, which is accompanied by astrocytic swelling. Cl-/HCO3- anion exchange inhibitors, such as SITS (4-acetamido-4'-isothiocyanostilbene-2,2' disulfonic acid) or furosemide, but not the specific cotransport inhibitor bumetanide, inhibit swelling or increased ion uptake in rat brain slices caused by elevated potassium although there were marked species differences in sensitivity. A novel anion exchange inhibitor, L-644,711, inhibits swelling and increased ion uptake caused by glutamate in rat and cat brain slices, as well as inhibiting [3H]glutamate uptake in primary rat astrocyte cultures. Possible mechanisms of action of the inhibitors are discussed. L-644,711 was also found to be effective in promoting recovery from a trauma plus hypoxia head injury model in cats. Marked perivascular astrocytic swelling is associated with this head injury model, and L-644,711 also inhibited such astroglial swelling as determined ultrastructurally. The significance of these findings in relation to possible connections between astrocytic swelling and brain pathology is discussed. PMID- 2570585 TI - The genetically epilepsy-prone rat. A valuable model for the study of the epilepsies. AB - In order to develop a rational clinical treatment for any pathological state, the molecular bases for that state must be understood. As simple and logical as that statement appears, it remains the major obstacle to effective treatment of the family of neurological disorders collectively called the epilepsies. Under the term, the epilepsies are grouped as several types of seizure processes that undoubtedly have multiple pathophysiological causes. Thus, the search to elucidate the molecular bases for the epilepsies has as one of its fundamental components the careful selection of an appropriate model system. The search for an "ideal" seizure model has essentially followed two paths. In the first, animals are rendered "epileptic" by artificial methods and then the pathophysiological, electrophysiological, and pharmacological changes are evaluated. In the second, animals are developed with a genetic predisposition to seizures and used to evaluate the molecular bases for the seizure-prone state. Work using both types of models have provided valuable information about the epileptic state. This review describes an epilepsy model developed using the second approach, namely, the Genetically Epilepsy-Prone Rat (GEPR). These animals represent a valuable model for the study of the inborn neurological defect that predisposes these animals to seizures. A brief description of the work done in several laboratories characterizing the model is presented. Finally, the value of the GEPR as a model for studying the pathophysiology of the epilepsies will be described. PMID- 2570586 TI - Hormonal modulation of the rat mammary gland gamma-glutamyltranspeptidase. AB - 1. The in vivo effect of estradiol and domperidone (a hypophysis stimulator of prolactin secretion) in immature ovariectomized-adrenalectomized and hypophysectomized rat mammary gland was studied. 2. gamma-Glutamyltranspeptidase activity was used to evaluate the role of estradiol in the specific response of the gland to prolactin. 3. Our results suggest that the activity of gamma glutamyltranspeptidase, like casein and lactose synthetase, is part of the specific response of the gland to prolactin. PMID- 2570587 TI - Effects of H2-blockers on rat mesenteric arterioles under resting conditions. AB - H2-receptor but not H1-receptor antagonists, administered by intravenous infusion, produce a dose-dependent constriction of superior mesenteric arterioles of the anaesthetized rat under resting conditions. However the possibility that this effect could be related to a blockade of H2 receptors is unlikely, since their potency on blood flow changes does not parallel known H2-receptor antagonist activity. Furthermore, the estimated potency ratio is not in the expected order if the vasoconstriction is due to H2-antagonism, the potency ratios on guinea-pig atrial muscle and on mouse gastric acid secretion being famotidine greater than oxmetidine greater than ranitidine greater than cimetidine. PMID- 2570588 TI - Interaction of non-piliated Neisseria gonorrhoeae strain 7122 and protein IA with an epithelial cell monolayer. AB - Studies with [14C] uracil-labelled bacteria revealed that the interaction of Neisseria gonorrhoeae with epithelial cells occurred in a time-dependent reaction which is slightly pH-dependent and optimal at pH 6.5. Immunofluorescence tests and immunoelectron microscopy of ultrathin sections confirmed the attachment of these bacteria to the epithelial cell membrane. The interaction of purified protein I with epithelial cells was time-dependent and reached equilibrium after four hours as shown by tracer experiments with 125I-labeled protein I. Cleavage experiments with trypsin followed by SDS-PAGE and autoradiography indicated that protein I (labeled with 125I) was associated with the membrane of the epithelial cells and only partly accessible by trypsin after its interaction with these mammalian cells. Immunofluorescence tests as well as immunoelectron microscopy with the monoclonal antibody G7A2C and gold-labeled protein A confirmed a dense association pattern of protein I with the cell monolayer. PMID- 2570589 TI - Subsets of rat CD4+ T cells express different variants of the leukocyte-common antigen: functions and developmental relationships of the subsets. PMID- 2570591 TI - Antigen processing: current issues, exceptional cases (Thy 1 alloantigen, MHC class-II-restricted cytolytic T cells), and implications for vaccine development. AB - A dogmatic view of antigen processing is presented in outline, followed by a survey of unresolved issues in the subject. The activity of Thy 1 as an alloantigen, and allospecific MHC Class-II-restricted cytolytic T cells offer examples of exceptional cases of antigen presentation. Implications for the design of vaccines are drawn. PMID- 2570590 TI - T-cell clone anti-clone interactions. Effects on suppressor and helper activities. AB - An experimental model of two interacting clones of T cells is described, which may be used for defining and exploring the T-cell immunoregulatory network. Mx9/9 is a CD4 clone bearing an antigen receptor recognized by the Mx9 anti-V beta 8 monoclonal antibody (MoAb). Anti-V beta 8 MoAbs activate and induce cell proliferation of this clone. Autologous clones were raised against Mx9/9 cells using the peripheral blood mononuclear (PBM) cells of the Mx9/9 clone donor (PBMjm). Some of these cloned anti-clone cells proliferated after stimulation with irradiated Mx9/9 cells, but not after stimulation with other autologous cloned T cells or heterologous PBM, suggesting that these clones recognize the T cell receptor (TCR) of the Mx9/9 cells. The proliferation of the Mx9/9 stimulated cloned anticlone cells was blocked by anti-class II MoAbs, indicating that the autoreactive clones recognize their target antigen in conjunction with HLA Class II products. The ability of clone Mx9/9 to proliferate after stimulation with anti-V beta 8 MoAb was inhibited when clone 121 cells were added to the cultures. However, clone 121 lost its suppressor function after 4 months in culture and instead gained the ability to enhance the proliferation of Mx9/9 cells in the presence of anti-V beta 8 MoAb. In contrast, clone 18 lacked suppressor activity at the early stage of the study but later acquired this function. We conclude that some autoreactive clones are not fully committed and may express more than a single function. Such cells cannot therefore be designated as 'suppressor cells', although they expressed suppressor potential at certain stages. PMID- 2570592 TI - Synthesis and dopamine receptor affinities of 6-amino-5,6,7,8-tetrahydroquinoline derivatives. AB - Some N,N-dialkylderivatives of 6-amino-5,6,7,8-tetrahydroquinoline were synthesized. The affinity of new compounds for dopamine binding sites was measured in a test involving displacement of [3H]SCH 23390 (D-1 selective) and [3H]spiperone (D-2 selective) from homogenized rat striatal tissue. While no compound was effective in displacing [3H]SCH 23390, in the binding assays on the D-2 receptor all tetrahydroquinolines displaced [3H]spiperone from specific binding sites, the compounds with a N-n-propyl-N-phenylethylamino group (18) or N,N-di n-propylamino group (16) being the most potent. PMID- 2570593 TI - Effects of benextramine on the adrenergic inhibition of insulin secretion in isolated rat pancreatic islets. AB - Insulin secretion from isolated rat islets of Langerhans in the presence of 4 mM glucose averaged 2.26 +/- 0.20 (S.E.M.) ng/islet per 90 min and was significantly (P less than 0.001; n = 30) increased to 3.28 +/- 0.21 ng/islet per 90 min by the covalent alpha-adrenoceptor antagonist benextramine (10 microM). Glucose (20 mM) also increased the secretion rate (to 6.24 +/- 6.0 ng/islet per 90 min) but, under these conditions, the response was not further enhanced by benextramine. Clonidine and noradrenaline (1 nM-10 microM) each caused dose-dependent inhibition of glucose-induced insulin secretion which was maximal at 1 microM. Benextramine, when added simultaneously with the agonist, relieved, in a dose dependent manner, the inhibition of secretion induced by either clonidine or noradrenaline with similar sensitivity. Even after a 30-min preincubation with benextramine the antagonist failed to differentiate between noradrenaline, adrenaline and clonidine with respect to inhibition of insulin secretion. In contrast to its effects on adrenergic responses, short-term treatment with benextramine did not significantly affect muscarinic-cholinergic receptor mediated 45Ca2+ efflux from rat islets of Langerhans perifused in Ca2+-depleted medium. These data suggest that benextramine does not differentiate between clonidine and noradrenaline in rat islets of Langerhans but that it does show preference for alpha-adrenoceptors in this tissue. PMID- 2570595 TI - The galactopharmacopedia. Narcotic analgesics: use in the breastfeeding woman. PMID- 2570594 TI - C4A gene deletion: association with Graves' disease. AB - The association of HLA class I and class II antigens, particularly HLA-B8,DR3, with a variety of autoimmune diseases has been well documented. The C4A*Q0 (non expressed C4A) allele which is in linkage disequilibrium with HLA-B8,DR3 has also been reported to be associated with systemic lupus erythematosus, insulin dependent diabetes mellitus and Graves' disease. However, the number of studies has been limited by the requirement of family data for the assignment of the C4A*Q0 allele based on C4 protein typing. Recently, with the availability of a C4 cDNA probe, a C4A gene deletion associated with HLA-B8,DR3 has been reported in normal individuals. We have tried to resolve the problem of assigning the C4A*Q0 allele by using both phenotypic and genotypic approaches and have determined the significance of the C4A*Q0 allele in 80 unrelated patients with Graves' disease and in 50 normal control subjects. Our results demonstrate a strong association of the C4A*Q0 allele with Graves' disease (56 versus 26%; P less than 0.002, relative risk = 3.7) and in particular in association with HLA-B8 and/or DR3 (92 versus 70.6%; P less than 0.04) when compared with normal controls. All the C4A*Q0 alleles that were associated with HLA-B8 and/or DR3 were due to a C4A gene deletion. Of the C4A*Q0 alleles, in Graves' disease, 94% (compared with 82% in the control group) could be detected by C4 DNA analysis using either TaqI or EcoRI restriction endonucleases. It is suggested that a combination of C4 protein typing with C4 DNA analysis is the best approach for the determination of the C4A*Q0 allele in unrelated individuals without access to family data. PMID- 2570596 TI - Immunogenetics of insulin-dependent diabetes mellitus in humans. AB - Diabetes mellitus is the second most prevalent chronic disease in children in the U.S. It is associated with severe manifestations which include blindness and circulation deficiencies as well as markedly increased risk of death. The etiology of diabetes mellitus remains a mystery although both genetic and environmental factors have been implicated. The geneticist is confronted with a number of obstacles in his attempts to unravel this problem, including differences in the definition of affected individuals. This matter was certainly clarified by the separation of noninsulin-dependent diabetes (NIDDM) and insulin dependent diabetes mellitus (IDDM) into two separate disease entities. Twin studies, however, show that IDDM cannot be entirely due to genetic causes as concordance is no more than about 50%. Although the disease is then clearly not inherited per se, the "susceptibility" to diabetes seems almost surely inherited and, provided this susceptibility, the disease can be brought on by environmental factors. Until the underlying mechanism causing IDDM is completely ascertained, we have to rely on genetic markers to approach the study of the inheritance thereof. Since, in the early 1970s, research by Nerup's and Cudworth's groups revealed associations between the HLA-B locus and IDDM, the HLA markers are considered the classical genetic markers for IDDM susceptibility. In this paper, we review the nature of the genetic susceptibility to IDDM and the possible environmental factors which can bring on the disease. PMID- 2570597 TI - Paraventricular nucleus lesions attenuate the development of hypertension in DOCA/salt-treated rats. AB - To determine whether paraventricular nucleus (PVN) can play a role in the hypertension in DOCA/salt-treated rats, DOCA/salt hypertension was produced in PVN lesions and sham-operated rats. In lesioned rats, the development of hypertension was significantly attenuated (day 7: 132 +/- 3 v 157 +/- 5 mm Hg, P less than 0.01; day 14: 132 +/- 3 v 157 +/- 5 mm Hg, P less than 0.01; day 21: 189 +/- 2 v 224 +2- 6 mm Hg, P less than 0.01). Lesions lowered systolic blood pressure in even control rats. Mean blood pressure (mBP) from awake free moving rats was also significantly lower in lesioned DOCA/salt-treated rats than those of sham-operated DOCA/salt-treated rats (155 +/- 14 mm Hg v 193 +/- 13, P less than 0.01), while mBP was not different between lesioned and sham-operated control rats. The reduction of mBP by hexamethonium injections was significantly larger in sham-operated DOCA/salt-treated rats than those of lesioned DOCA/salt rats. (-53 +/- 3% v -45 +/- 2, P less than 0.05). Plasma norepinephrine and epinephrine were significantly elevated in DOCA/salt-treated rats, however, PVN lesions inhibited significantly those elevations. 1-Deaminopenicillamine, 4 valine, 8-D-arginine Vasopressin (dPVDAVP) injections did not affect BP and heart rate in all rats. Body weight, water intake, urine volume, urine Na, K, and vasopressin excretion, and urine osmorality were not altered by lesions. These findings suggest that PVN contributes to development of hypertension in DOCA/salt treated rats with sympathetic nervous activations. PMID- 2570598 TI - D-1 dopamine agonist administration reduces the threshold for convulsions produced by pilocarpine. AB - Focal, limbic seizures were produced by systemically administered pilocarpine (200 mg/kg, i.p.); as previously described this dose produces limbic stereotypies but neither convulsions nor seizure-related brain damage. The pretreatment, 5 minutes prior pilocarpine, with the D-1 agonist SKF 38393 (-ED50 = 1 mg/kg; i.p.) induced convulsions similar to those produced by a higher, convulsant dose of pilocarpine. On the other hand, the pretreatment with the D-2 agonist LY 171555 failed to induce convulsions. The D-1 receptor antagonist SCH 23390 prevented the convulsions induced by SKF 38393 plus pilocarpine (200 mg/kg). This study indicates that D-1, but not D-2, receptor stimulation converts subconvulsant doses of pilocarpine into convulsant ones. PMID- 2570599 TI - Further observation on the intraepithelial nerve fibers of rabbit ocular ciliary epithelium. AB - The intraepithelial nerve fibers of rabbit ciliary epithelium were observed under the electron microscope after administration of 5-hydroxydopamine. All the fibers so far observed contained small and large cored vesicles, thus indicating their noradrenergic nature. PMID- 2570600 TI - Somatostatin-containing neurons in the mouse brain: an immunohistochemical study and comparison with the rat brain. AB - The distribution of somatostatin-containing neurons in mice of both sexes was immunohistochemically examined and compared with that in rats. In radioimmunoassay the relative somatostatin content in the mouse brain was 2-3 times higher than that in the rat. The overall immunohistochemical staining for somatostatin was much stronger and more prominent in the mouse than in the rat. Although the distribution pattern of somatostatin immunoreactivity was basically the same between the two animals, several regions, especially the nucleus anterior hypothalami and the nucleus interpeduncularis, were found to contain large aggregates of somatostatin-immunoreactive neurons in the mouse brain but not in the rat. The electrolytic lesions to the nucleus anterior hypothalami caused a marked decrease in somatostatin immunoreactivity of the outer layer of the median eminence in the mouse. This suggests that the nucleus anterior hypothalami is an additional source of somatostatin for the median eminence in the mouse. The differences recognized between the species are interesting from functional and evolutionary points of view. PMID- 2570601 TI - Assessment of airways, tremor and chronotropic responses to inhaled salbutamol in the quantification of beta 2-adrenoceptor blockade. AB - 1. The purpose of the study was to assess and compare the effects of inhaled salbutamol on heart rate (HR), finger tremor (Tr) and specific airways conductance (sGaw) in the measurement of beta 2-adrenoceptor blockade in normal subjects. 2. Five healthy volunteers were given oral doses of atenolol 50 mg, 100 mg, 200 mg (A50, A100, A200), propranolol 40 mg (P40) or identical placebo (P1) in a single-blind crossover design. 3. Three hours after drug ingestion, dose response curves were constructed using cumulative doses of inhaled salbutamol: 200 micrograms, 700 micrograms, 1700 micrograms, 3200 micrograms, 6200 micrograms. HR, Tr and sGaw were measured at each dose increment, made every 20 min. 4. Increasing doses of atenolol were associated with progressive reduction in salbutamol induced beta-adrenoceptor responses. The greatest attenuation occurred with propranolol. These effects on beta-adrenoceptor responses were similar for HR, Tr and sGaw. Geometric mean dose ratios (compared with placebo) for A50, A100, A200 and P40 were as follows HR: 1.98, 2.75, 4.29; Tr: 1.60, 3.78, 6.34, 80.50; sGaw: 1.08, 4.35, 12.30, 66.0 (no dose ratio was obtained for HR with P40). 5. These results showed that atenolol and propranolol attenuated the effects of salbutamol on HR to a similar degree as Tr and sGaw. Furthermore, the variability was least in the measurement of chronotropic responses, suggesting that this may be used to quantify beta 2-adrenoceptor antagonism. The beta 1 adrenoceptor selectivity of atenolol was a dose-dependent phenomenon, although the beta 2-adrenoceptor blockade of A200 was much less than with P40. PMID- 2570602 TI - Neuromuscular blockade with vecuronium in paediatric patients with burn injury. AB - 1. The effect of body surface area (BSA) burn injury on neuromuscular pharmacodynamics of vecuronium was evaluated. 2. The neuromuscular responses of 15 burned children were compared with those in five controls. 3. The effective dose for 50% suppression of twitch tension (ED50) was 34 micrograms kg-1 for patients with less than 40% BSA burn, 55 micrograms kg-1 for 40-60% BSA burn, and 65 micrograms kg-1 for patients with greater than 60% BSA burn. These values were significantly higher than the value for control patients, which was 18 micrograms kg-1. 4. Burn injury induces a resistance to the neuromuscular effects of vecuronium, the magnitude of which is related to burn size. PMID- 2570604 TI - Neuroleptanesthesia versus thoracic epidural anesthesia for abdominal aortic surgery. AB - The hemodynamic consequences of abdominal aortic surgery with infrarenal cross clamping were studied in 21 patients randomized in two groups. In Group I (11 patients), neuroleptanesthesia was utilized, while Group II (10 patients) received thoracic epidural anesthesia at the T8-9 level. Hemodynamic measurements were performed using Swan-Ganz catheters during the surgical procedures in all patients, with special attention to the periods of clamping and unclamping of the abdominal aorta. The thoracic epidural anesthesia group was characterized by greater hemodynamic stability during surgery, while patients in the neuroleptanesthesia group had significant lability of blood pressure, heart rate, and cardiac index. Nevertheless, in the two groups of patients, it is suggested that cardiac function was unfitted to the tissue oxygen demand after unclamping of the aortic prosthesis because the saturation in oxygen of the mixed venous blood and an increase in arteriovenous difference in oxygen were documented. These results point out that, whatever the anesthesia technique, the critical period in abdominal surgery could be aortic unclamping. PMID- 2570603 TI - The effect of roxatidine acetate and cimetidine on hepatic drug clearance assessed by simultaneous administration of three model substrates. AB - The effect of pretreatment for 7 days with either roxatidine acetate 75 mg twice daily or cimetidine 200 mg four times daily on the kinetics of antipyrine (AP), trimethadione (TMO) and indocyanine green (ICG) was studied in seven healthy, male, nonsmoking subjects. After pretreatment with cimetidine, the plasma clearances (CL) of AP and TMO were significantly lower and the elimination half life (t1/2) of AP was significantly increased. The volumes of distribution (V) of AP and TMO were not affected. After roxatidine acetate, the pharmacokinetics of AP and TMO were unchanged. The cumulative renal excretion (% dose) and formation clearance of 3-hydroxymethyl-3-nor-antipyrine (NORA) were lowered by cimetidine treatment, but not following the administration of roxatidine acetate. ICG clearance was not changed significantly by either pretreatment. The results of this study show that roxatidine acetate does not impair the metabolism of three model substrates used to assess hepatic drug clearance. PMID- 2570606 TI - Commentary on 'Crystallization of Michaelis Complex of D-Amino Acid Oxidase'. PMID- 2570605 TI - Chemical events in chloropropionyl coenzyme A inactivation of acyl coenzyme A utilizing enzymes. AB - Incubation of 3-chloropropionyl-CoA with 3-hydroxy-3-methylglutaryl-CoA synthase results in exchange of the C2 proton with solvent as inactivation of enzyme proceeds. This enzyme is also inhibited by S-acrylyl-N-acetylcysteamine; the limiting rate constant for inactivation by the acrylyl derivative (0.36 min-1) slightly exceeds the value measured for chloropropionyl-CoA (0.31 min-1). These observations support the intermediacy of acrylyl-CoA in the chloropropionyl-CoA dependent inactivation of hydroxymethylglutaryl-CoA synthase. Inhibition of fatty acid synthase by chloropropionyl-CoA is primarily due to alkylation of a reactive cysteine, although secondary reaction with the enzyme's pantetheinyl sulfhydryl occurs. Modification of fatty acid synthase by S-acrylyl-N-acetylcysteamine occurs at a limiting rate (1.8 min-1) that is comparable to that estimated for chloropropionyl-CoA-dependent inactivation. However, this enzyme lacks the ability to deprotonate C2 of an acyl group such as the chloropropionyl moiety. Since such a step would be required to generate an acrylyl group from chloropropionyl-S-enzyme, it is likely that a typical affinity labeling process accounts for inactivation of fatty acid synthase by chloropropionyl-CoA. HMG-CoA lyase is also inhibited by S-acrylyl-N-acetylcysteamine. In contrast to the ability of this reagent to serve as a mechanism-based inhibitor of hydroxymethylglutaryl-CoA synthase and an affinity label of fatty acid synthase, it acts as a group-specific reagent in modifying HMG-CoA lyase (kappa 2 = 86.7 M 1 min-1). PMID- 2570607 TI - Crystallization of Michaelis complex of D-amino acid oxidase. 1962. PMID- 2570608 TI - NADH-sensitive propionyl-CoA hydrolase in brown-adipose-tissue mitochondria of the rat. AB - Acyl-CoA hydrolase activity was studied in brown adipose tissue (BAT) mitochondria of rats. The substrate specificity was investigated: total hydrolase activity showed two activity peaks, one sharp peak for propionyl-CoA and a broad peak at medium- to long-chain acyl-CoAs. The propionyl-CoA activity fully comigrated with a mitochondrial matrix marker enzyme in fractionation studies of tissue and mitochondria. The hydrolytic activity against short-chain acyl-CoAs was inhibited by NADH, and analyses of the substrate specificity of the hydrolases in the presence and absence of NADH allowed for the delineation of two distinct acyl-CoA hydrolases. These hydrolases could also be separated by gel filtration. It was concluded that rat BAT mitochondria possess at least two matrix acyl-CoA hydrolases: one broad-spectrum acyl-CoA hydrolase with an apparent native molecular weight of less than 100,000, and a specific propionyl CoA hydrolase with an apparent native molecular weight at least 240,000; this hydrolase is regulated by NADH. It is suggested that the function of the propionyl-CoA hydrolase is to ensure that the level of propionyl-CoA in the mitochondria is not detrimentally increased. PMID- 2570609 TI - Effects of bradykinin on prostaglandin synthesis and cytosolic calcium in rabbit subcultured renal cortical smooth muscle cells. AB - Smooth muscle cells were cultured from an arteriole-rich fraction of the rabbit renal cortex and characterized by their ultrastructural and immunohistochemical features, their high content in creatine kinase (60-times that of the initial preparation) and their ability to synthesize renin. Cells, studied between passages 2 and 5, produced mainly PGE2 and, to a lesser extent, PGF2 alpha. Bradykinin (BK) (0.1 nM-1 microM) induced a concentration-dependent increase in PGE2 (28-40-times basal value at 1 microM after a 5 min incubation period) and stimulated also the free cytosolic calcium concentration [( Ca2+]i) with a 2-fold maximal rise to its basal value. Both effects, inhibited by the anti-B2 receptor [Thi5.8D-Phe7] BK, were not reproduced by DesArg9 BK. A decrease in the extracellular calcium concentration and incubation in the presence of a calcium channel blocker (lanthanum chloride) inhibited the BK-dependent rise of [Ca2+]i but not that of PGE2. Preincubation with phorbol myristate acetate increased basal and BK-induced PGE2 synthesis but prevented the effect of BK on [Ca2+]i. These results demonstrate the ability of BK to increase [Ca2+]i and PGE2 production in cultured vascular cells from the rabbit renal cortex and suggest that kinins might act on the cortical microcirculation via their direct effects on arteriolar smooth muscle cells. PMID- 2570612 TI - Drugs from emasculated hormones: the principle of syntopic antagonism. AB - This lecture outlines the early stages in the discovery of adrenaline beta receptor antagonists and of the histamine H2-receptor antagonists. It ends with a brief personal view about future research. PMID- 2570610 TI - Co-induction by peroxisome proliferators of microsomal 1 acylglycerophosphocholine acyltransferase with peroxisomal beta-oxidation in rat liver. AB - Administration of clofibric acid, 2,2'-(decamethylenedithio)diethanol, di(2 ethylhexyl)phthalate or perfluorooctanoic acid to male rates increased markedly microsomal 1-acylglycerophosphocholine (a-acyl-GPC) acyltransferase in a dose dependent manner in liver. Simultaneous administration of actinomycin D or cycloheximide completely abolished the increase in the enzyme activity. The treatment of rats with clofibric acid did not affect the rate of decay of 1-acyl GPC acyltransferase. Regardless of a great difference in the chemical structures of the peroxisome proliferators, high correlation was observed between the induced activities of microsomal 1-acyl-GPC acyltransferase and peroxisomal beta oxidation. Stearoyl-CoA desaturase was induced by peroxisome proliferators in a dose-dependent manner; nevertheless, high correlation was not seen between the induced activities of desaturase and peroxisomal beta-oxidation. Hormonal (adrenalectomy, diabetes, hyperthyroidism and hypothyroidism) and nutritional (starvation, starvation-refeeding, fat-free diet feeding and high-fat diet feeding) alterations hardly affected the activity of 1-acyl-GPC acyltransferase. The present results indicate that microsomal 1-acyl-GPC acyltransferase is a useful parameter responsive to the challenges by peroxisome proliferators and suggest that a similar regulatory mechanism operates for the inductions of microsomal 1-acyl-GPC acyltransferase and peroxisomal beta-oxidation. PMID- 2570611 TI - Biosynthesis, processing and half-life of P-glycoprotein in a human multidrug resistant KB cell. AB - The biosynthesis, processing, and half-life of the drug efflux pump, P glycoprotein, were studied in human multidrug-resistant KB (KB-C2) cells selected for resistance to colchicine. An antibody directed against a synthetic oligopeptide corresponding to the amino-acid sequence (Glu-393-Lys-408) of P glycoprotein from human mdr1 cDNA was prepared in rabbits. With immunoblotting and immunoprecipitation, we detected a 140-170 kDa protein in KB-C2 cells but not in parental sensitive KB cells. KB-C2 cells made a 125 kDa precursor that was slowly processed (t1/2 = 45 min) to the mature form of 140-150 kDa. The processing rate of P-glycoprotein was slower than that of low-density lipoprotein receptor. We detected another 160-180 kDa smear band, which might be a completely denatured form of P-glycoprotein. With immunoblotting, a minor band of high molecular mass (greater than 500 kDa) was also detected and this form increased after the cells were treated with chemical cross-linker, 1,5-difluoro-2,4 dinitrobenzene. The half-life of P-glycoprotein was long; no significant loss of P-glycoprotein was observed within 24 h after synthesis. Cells treated with tunicamycin produced a 120 kDa form of P-glycoprotein which was no longer processed but showed stability similar to that of the mature 140-150 kDa form. Agents that reverse multidrug resistance, phorbol ester and transport substrate did not affect the stability of P-glycoprotein. PMID- 2570613 TI - [Why is it sometimes necessary to include a large number of patients in clinical trials?]. PMID- 2570614 TI - Spastic dysphonia and denervation signs in a young man with tardive dyskinesia. AB - Pathophysiological theories of tardive dyskinesia (TD) suggest the possibility of structural changes in the central nervous system of patients with TD. This report describes a case of choreoathetoid dyskinesia and spastic dysphonia associated with clinical and electromyographic signs of muscle denervation. The findings of this case suggest that the neurological syndrome originates within basal ganglia nuclei but may also extend to the peripheral neuromuscular system. PMID- 2570615 TI - Potassium-induced release of endogenous glutamate and two as yet unidentified substances from the lateral line of Xenopus laevis. AB - The release of endogenous glutamate and other primary amines from the lateral line of Xenopus laevis was studied using an in vitro superfusion technique and high performance liquid chromatography. Potassium stimulation (50 mM KCl) applied to 60 or 120 lateral-line organs dissected from the skin and pooled in a perfusion chamber induced the release of glutamate and aspartate. The release of aspartate was smaller than that of glutamate and more variable. A variable release of two, as yet, unidentified substances was also detected. In low calcium (0.1 mM CaCl2), high magnesium (10 mM MgCl2) solution, 50 mM potassium failed to induce an increase in glutamate, aspartate and the two unknowns, suggesting they are released in a transmitter-like manner. The technique presents a new and simple method for studying transmitters in hair-cell systems. Although other interpretations are possible, the results are consistent with the hypothesis that glutamate is a hair-cell transmitter and suggest a potential role for other substances in the transduction process, perhaps as neuromodulators. PMID- 2570616 TI - NMDA antagonists and potentiation of NMDA-induced motoneuron depolarizations in the isolated frog spinal cord. AB - The action of N-methyl-D-aspartate (NMDA) antagonists on motoneurons was studied in the isolated, hemisected frog spinal cord using sucrose gap techniques. NMDA evoked motoneuron depolarizations were depressed by application of APV, APH, kynurenate, Mg2+ ions, ketamine, and MK-801. Upon returning to normal Ringer's solution after exposure to all antagonists (except MK-801). NMDA responses were significantly potentiated. Kainate- and quisqualate-induced depolarizations were unchanged. The facilitation appeared to result, at least in part, from a direct action on motoneuron membranes since it persisted in the presence of tetrodotoxin which eliminated interneuronal firing. However, indirect actions involving interneurons also contributed to the potentiation because NMDA-evoked changes in K+ release were increased following exposure to NMDA antagonists and return to normal medium. Reduction of temperature (7 degrees C) which should reduce amino acid uptake did not affect results with APV. In addition, desensitization of NMDA responses was not altered by application of APV. The results indicate that NMDA antagonists have complex and long-lasting effects on the function of the NMDA receptor complex. PMID- 2570617 TI - Antinociceptive effect of agonists microinjected into the anterior pretectal nucleus of the rat. AB - Electrical stimulation at many sites within the pretectal complex and adjacent structures of the rat dorsomedial thalamus yields antinociception. It is documented that no other site in this region evokes antinociception longer lasting than that obtained by stimulation of the anterior pretectal nucleus (APtN). The effects of agonists injected into different nuclei of the dorsomedial thalamus on the tail-flick reflex of rats in response to noxious heat was examined. All animals were submitted to intracerebral electrical stimulation and microinjection of agonists. It was confirmed that strong and long lasting antinociception followed brief (15 s), low intensity (35 microA rms) stimulation of the APtN. In addition, L-glutamate (3.5 and 7.0 micrograms), morphine (1.0 and 5.0 micrograms), and 5-hydroxytryptamine (5-HT; 2.5 and 5.0 micrograms), but not acetylcholine (5.0 micrograms), carbachol (2.5 micrograms), norepinephrine (5.0 micrograms), or dopamine (5.0 micrograms), induced dose-dependent antinociception when microinjected into the APtN. The effect of 5-HT was fully depressed by pretreating animals with methysergide (5 mg/kg, i.p.). A survey of the sites from which morphine and 5-HT induce antinociception revealed that in no site of the dorsomedial thalamus did the effect last longer than after microinjection into the APtN. It is concluded that antinociception evoked by stimulation of the APtN depends on the activation of neuronal cell bodies in the nucleus, and that 5-HT and endogenous opioids may play a physiological role as neurotransmitters mediating antinociception in the APtN. PMID- 2570618 TI - One year treatment with haloperidol or clozapine fails to alter neostriatal D1- and D2-dopamine receptor sensitivity in the rat. AB - Rats were treated continuously with either haloperidol (HAL), clozapine (CLOZ) or tap water for one year. There were no differences between age-matched control and antipsychotic drug (APD) treated groups regarding the effects of the D1-agonist (+)-SKF 38393 or the D2-agonist quinpirole on striatal cAMP content. However, the combination of SKF (10 microM) and quinpirole (1 microM) produced a marked synergistic response in HAL-treated animals as compared to controls. Our data fail to support the hypothesis that APD produce their neurological side effects by inducing D2-receptor hypersensitivity in the basal ganglia. However, the results do suggest that chronic APD treatment alters the interaction between D1- and D2-neostriatal receptors. PMID- 2570619 TI - An in vivo procedure for the selective inactivation of rat brain cerebral cortical alpha-adrenoceptors using N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ). AB - Intraperitoneal administration of 0.8 mg/kg N-ethoxycarbonyl-2-ethoxy-1,2 dihydroquinoline (EEDQ) to rats significantly reduced alpha-adrenoceptor binding in cerebral cortical membranes without affecting beta-adrenoceptor sites. A selective reduction in alpha 1-adrenoceptors was achieved by injecting yohimbine prior to EEDQ, whereas prazosin pretreatment yielded a selective reduction in alpha 2-adrenoceptor binding. Administration of EEDQ decreased norepinephrine stimulated inositol phosphate and cyclic adenosine monophosphate (cAMP) accumulation in cerebral cortical tissue as well as the cAMP response to isoproterenol in combination with 6-fluoronorepinephrine without modifying the second messenger response to isoproterenol alone. The results suggest that, under the proper conditions, EEDQ administration can selectively diminish rat brain alpha-adrenoceptor number and function, yielding a procedure that may be useful for defining the behavioral and physiological properties of these sites. PMID- 2570620 TI - Vagally mediated insulin secretion by stimulation of brain cholinergic neurons with neostigmine in bilateral adrenalectomized rats. AB - This study investigated the relationship between central cholinergic neurons and insulin secretion in bilateral adrenalectomized fed rats. Neostigmine (a cholinesterase inhibitor, 5 x 10(-8) mol) administered into the third cerebral ventricle produced significant increases in hepatic venous plasma insulin and glucose concentrations, whereas i.v. injection of the same dose of neostigmine did not. Prior acute subdiaphragmatic vagotomy or i.p. pre-injection with methylatropine (10(-8) mol) completely prevented the neostigmine-induced rise in plasma insulin concentration. Intraperitoneal pretreatment with hexamethonium (5 x 10(-8) mol) also significantly reduced the plasma insulin response. These peripheral pretreatments did not change the plasma glucose response to neostigmine. Intraventricular co-administration of 10(-9) mol methylatropine, a dose that was ineffective when pre-injected i.p., eliminated the plasma insulin and glucose responses to neostigmine, whereas hexamethonium (5 x 10(-8) mol) had no influence on either response to neostigmine. These observations suggest that stimulation of central cholinergic-muscarinic neurons with third cerebral ventricular injection of neostigmine results in vagally mediated insulin secretion in bilateral adrenalectomized fed rats. PMID- 2570621 TI - Endogenous opioid system down-regulation during hibernation in amphibians. AB - In late summer, Northern grass frogs, Rana pipiens, intraspinally administered dynorphin produces a potent, dose-dependent antinociceptive action as measured by the acetic acid test used to evoke a hindlimb wiping response. Surprisingly, in fall, frogs which have entered hibernation, intraspinal dynorphin produces no antinociception action. Intraspinal morphine shows a decreased effect in fall frogs while systemic morphine is equi-effective in summer and fall frogs. Immobilization stress, previously shown to be mediated by endogenous opioid systems in this amphibian, produces a robust increase in nociceptive thresholds in summer frogs while the nociceptive thresholds of fall frogs are unaffected by this procedure. Summer frogs adapted to cold room (4 degrees C) show a significant decrease in nociceptive thresholds compared to cohorts kept at room temperature, and cold-adapted frogs returned to room temperature show a naloxone attenuated increase in nociceptive threshold. Collectively, these data suggest that endogenous opioid systems in these northern frogs are down-regulated during fall hibernation. PMID- 2570622 TI - Somatostatin-, vasoactive intestinal polypeptide- and neuropeptide Y-like immunoreactivity in eye- and submandibular gland-projecting sympathetic neurons. AB - Studies combine the use of the retrograde tracer, fluorogold, and immunocytochemical staining to determine whether superior cervical ganglion (SCG) neurons projecting to the iris or submandibular gland (SMG) in adult male and female rats show distinctive immunoreactivity to somatostatin (SS), vasoactive intestinal polypeptide (VIP), or neuropeptide Y. Overall, more SMG-projecting neurons than eye-projecting neurons contain VIP-like immunoreactivity (VIP-LI), and more eye-projecting neurons than SMG-projecting neurons contain SS-LI and VIP LI. Thus, postganglionic neurons of the SCG that project to specific target tissues are heterogeneous in their peptide content, and there are differences in the pattern of peptide-immunoreactivity between neurons projecting to these two target tissues. In addition, the results indicate that there may be gender differences in the expression of these neuropeptides. PMID- 2570623 TI - The cholinergic influence upon rat dorsal lateral geniculate nucleus is dependent on state of arousal. AB - Several lines of evidence suggest a role for acetylcholine (ACh) in mediating the effects of state of arousal on transfer of visual information through the lateral geniculate nucleus (LGN). Local application of cholinergic agonists to geniculate relay cells in anesthetized cats and rats produces predominantly facilitatory effects. This indicates that presynaptic release of ACh may be responsible for the increased excitability of LGN relay cells that is observed during waking and REM sleep. In this study in rats we have examined the influence of cholinergic agonists applied during the 3 natural states of arousal: waking, slow-wave (SW) sleep and rapid eye movement (REM) sleep. Pharmacological agents were iontophoretically administered to identified, single cells in head-restrained, unanesthetized rats free to sleep and wake. Application of cholinergic agonist produced state-dependent differences in response in all geniculate relay-cells studied. During both waking and REM sleep, a facilitatory response was always observed, whereas in SW sleep responses were of three types: no effect (62%), inhibition (24%), and biphasic inhibition followed by facilitation (14%). All response types were antagonized by scopolamine. In contrast to the qualitatively different state-dependent effects of cholinergic agonists, response to application of glutamate, with quantitative variations, was uniformly facilitatory in all states, though responses in SW sleep tended to be lower in magnitude. The effects of gamma-aminobutyric acid (GABA), glycine, and serotonin were inhibitory in all states. These data are consistent with the suggested role of ACh in mediation of increased relay-cell excitability during REM sleep and waking. Our findings, however, also indicate that in the transition from SW sleep to REM or waking, local release of ACh is not solely responsible for alterations in cell excitability. PMID- 2570624 TI - Brain amino acid concentrations and Ca2+-dependent release in intractable depression assessed antemortem. AB - The concentrations of 3 putative neurotransmitters (glutamate, aspartate and gamma-aminobutyrate), 4 related amino acids and 5 non-transmitter-related amino acids have been measured in neurosurgical samples (frontal cortex) from patients with intractable depression and controls. In addition, the glutamate receptor agonist 2-amino-4-sulpho-butanoic acid (homocysteic acid) has been identified in human brain and measured in these samples. There were no changes in the concentrations of amino acids in depressed patients compared to control with the exception of aspartic and homocysteic acids which were elevated in a sub-group of patients with depression compared to control. The Ca2+-dependent release (K+ stimulated) of putative neurotransmitters has been demonstrated for the first time from brain tissue of depressed patients. Glutamate release was unaltered from the control value. Aspartate values showed unexplained variability but it's release and that of gamma-aminobutyrate were elevated in some depressed subjects. These results do not support the hypothesis of reduced amino acid function in depressive illness. PMID- 2570625 TI - L(+)-2-amino-4-phosphonobutyrate inhibits the release of both glutamate and dynorphin from guinea pig but not rat hippocampal mossy fiber synaptosomes. AB - The K+-evoked release of dynorphin A(1-8)-like immunoreactivity from guinea pig hippocampal mossy fiber synaptosomes was inhibited 53% by L(+)-2-amino-4 phosphonobutyrate (L(+)APB, 300 microM), a glutamate analogue. Equimolar L(+)APB also inhibited the Ca2+-dependent component of endogenous L-glutamate release from these mossy fiber synaptosomes by 40%. The K+-evoked release of both glutamate and dynorphin A(1-8) from rat hippocampal mossy fiber synaptosomes were unaffected by L(+)APB. It is proposed that L(+)APB selectively suppresses the excitatory mossy fiber synaptic inhibiting the Ca2+-dependent release of glutamate and dynorphin A(1-8) from guinea pig but not rat hippocampal mossy fiber terminals. PMID- 2570626 TI - NMDA receptor activation by residual glutamate in glutamine preparations: a cautionary note regarding weak NMDA receptor agonists. AB - During an investigation of excitatory amino acids on cultured embryonic Xenopus neurons, we observed that commercial preparations of glutamine had weak agonist activity on NMDA-type glutamate receptors. Threshold responses were observed at 100 microM glutamine, and the dose-response relation did not show inflection or saturation at concentrations of up to 10 mM. However, NMDA receptor activation induced by glutamine probably represented activity of residual glutamate because: (a) recrystallization of glutamine reduced residual glutamate levels (measured by HPLC analysis) and NMDA receptor activation by comparable amounts; and (b) glutamate at concentrations close to those predicted to be present in glutamine preparations elicited currents of similar amplitudes. Our data indicate that residual glutamate at levels of less than 0.05% are sufficient to confound studies of weak NMDA receptor agonists. PMID- 2570628 TI - Spontaneous activity induced in isolated mouse spinal cord by high extracellular calcium and by low extracellular magnesium. AB - Spontaneous discharges were observed in recordings from dorsal and ventral roots (DRs and VRs) of hemisected mouse spinal cords in vitro when bath [Ca2+] was raised from the control level of 1.2 to 1.8 or 2.4 mmol/l, and when bath [Mg2+] was lowered resembling drug-induced 'interictal' discharges described earlier. Maximum discharge frequency was reached at 2.4 or 3.6 mmol/l [Ca2+] while at higher concentrations mean frequency diminished but mean amplitude still increased somewhat. The frequency distribution of wave amplitudes suggests recruitment of neurons by groups or assemblies. The pacemaker is located in the dorsal spinal quadrant. The GABAA receptor antagonists picrotoxin and bicuculline in low concentration blocked the spontaneous activity, indicating an obligatory GABAergic link in the pacemaker circuit. The NMDA antagonists D.L-APV and D-APV in high concentration variably depressed but did not abolish spontaneous activity. The propensity of spinal neuron assemblies for self-paced discharge may contribute to the pathologic irritability of injured spinal cords. PMID- 2570627 TI - Effects of ageing on the behavioural responses to dopamine agonists: decreased yawning and locomotion, but increased stereotypy. AB - Sensorimotor function and the behavioural responses to a range of doses of subcutaneous apomorphine were assessed in mature (6-8 months) and old (23-26 months) Sprague-Dawley rats of comparable weight. In addition, the locomotor activity response of 12-month-old and 24-month-old rats to continuous infusions (14 days by osmotic minipump) of a selective dopamine D2 agonist. (+)-4-propyl-9 hydroxynaphthoxazine (PHNO, 10 micrograms/h) was investigated. Measures of spontaneous locomotor activity and motor coordination revealed impairments in the aged animals. Low doses of apomorphine (10-50 micrograms/kg), which preferentially activate dopamine autoreceptors, induced yawning, chewing mouth movements and penile grooming. The frequency of yawning and duration of penile grooming were significantly decreased in the old animals. In contrast, 200 micrograms/kg of apomorphine induced stereotyped sniffing and licking or gnawing, and these responses were significantly increased in the aged animals. There was a 25% decrease in striatal dopamine levels in the aged animals in this experiment. PHNO increased the amplitude of the circadian rhythms in locomotor activity exhibited by mature rats, and daytime tolerance to the stimulant effects of PHNO was reversed by stress in these animals. Both of these effects were attenuated in the aged rats. These findings suggest that (1) the dopamine receptors mediating yawning and stereotypy have different anatomical locations (2) ageing is associated with decreased responsiveness to stimulation of dopamine autoreceptors, consequent upon the loss of dopaminergic nerve terminals, and (3) while the functional response to selective stimulation of postsynaptic D2 receptors decreases with age, the postsynaptic response to a mixed D1/D2 agonist increases. PMID- 2570629 TI - Modulatory effects of catecholamines on neurons of the rat visual cortex: single cell iontophoretic studies. AB - The catecholamines noradrenaline and dopamine have been proposed as neuromodulators of cortical neuron excitability, and such a regulation could be mediated by specific adrenergic and dopaminergic receptors. We characterized electrophysiologically some of the types of responses to the iontophoretic application of adrenergic and dopaminergic agonists and antagonists on single cells in the rat visual cortex (areas occipital 1 monocular or Oc1M and occipital 1 binocular or Oc1B). For the majority of spontaneously active and visual cortical cells, noradrenaline and dopamine decreased the firing frequency. In the case of visually driven (synaptically activated) neurons, background firing was the main component of the response to be inhibited by the administration of noradrenaline, clonidine, and oxymetazoline, leading to an enhancement of the signal-to-noise ratio. Since these effects could be reduced or blocked by a previous ejection of the specific alpha 2-antagonist idazoxan, the findings support a role for alpha 2-adrenergic receptors in the transmission of sensory inputs to the visual cortex. These effects were not found with the mixed alpha adrenergic agonist phenylephrine nor with the beta-agonist isoproterenol. Finally, the use of the inhibitory amino acid GABA rules out a simple hyperpolarizing response as the mechanism underlying noradrenaline modulatory effects in the cerebral cortex. PMID- 2570630 TI - Platelet-activating factor: lack of direct action on guinea pig myocardium and possible transmitter release from cardiac sympathetic nerve endings at high concentrations. AB - Microelectrode and mechanical studies were performed with isolated guinea pig myocardium (right ventricular free walls and papillary muscles) to examine the effects of platelet-activating factor (PAF) and lysophosphatidylcholine (LPC). Low concentrations of PAF (10(-8) to 10(-6) M, a range equivalent to the blood concentrations that produce marked hypotension in vivo) had no effects on action potential configuration and contractile force. High concentrations (10(-5) to 10( 4)M) of PAF and LPC per se elicited slow response action potentials with concomitant contraction (restored contraction) in the myocardium depolarized with elevated K+ (25 mM); they also augmented slow responses and restored contractions produced by a low concentration of isoproterenol (10(-8) M). Although these results suggested there was an increase in slow Ca current, the slow responses and restored contractions thus produced were greatly suppressed or abolished by the addition of a beta-adrenoceptor blocking agent, sotalol (10(-5) M), and by pretreatment with reserpine (5 mg/kg i.p., 24 h prior). In accordance with our previous conclusions, the present results suggest that direct cardiac action is not involved in the mechanisms of hypotension produced by PAF. It was also shown that high concentrations of PAF and LPC may act nonspecifically as amphiphilic compounds to induce transmitter release from sympathetic nerve endings, which may in turn augment the Ca current channels in the myocardial cell membrane. PMID- 2570631 TI - Cardioprotective effect of metipranolol on ischemic heart muscle. AB - The effect of the beta-adrenoceptor antagonist metipranolol given twice daily in a dose of 0.5 mg/kg body weight for 3 days on respiration, respiratory control index of mitochondria and oxidative phosphorylation was measured in the heart muscle of anesthetized dogs after 60 min of induced ischemia. In control animals pretreated with saline, all measured variables with the exception of the coefficient of oxidative phosphorylation were significantly decreased. In contrast, pretreatment with metipranolol significantly improved all measured myocardial metabolic variables. The effect of metipranolol on the oxidative phosphorylating processes was more pronounced when the metabolic substrate was glutamate than when it was pyruvate. PMID- 2570633 TI - Reactivity of monoclonal islet cell antibodies on normal hyperplastic and neoplastic thyroid C-cells. AB - Thyroid C-cell reactivity to 15 monoclonal antibodies raised against a series of pancreatic islet cells (H[human]ISL,B[bovine]ISL and R[rat]ISL) was evaluated using an indirect immunoperoxidase technique on frozen thyroid sections. Of the monoclonal anti-islet cell antibodies, five reacted specifically with bovine C cells or human hyperplastic and neoplastic C-cells but not with follicular cells. Two monoclonal antibodies of the bovine series showed strong immunoreactivity with C-cells and only a weakly positive immunostaining of follicular cells. Five monoclonal antibodies reacted with both thyroid C-cells and follicular cells, whereas 3 monoclonal anti-islet cell antibodies did not strain any cell type of the thyroid. In human medullary carcinomas, calcitonin- and somatostatin producing neoplastic cells were immunoreactive with the same monoclonal antibodies as were normal human C-cells. The protein bands identified by the monoclonal antibodies in human medullary carcinomas had the same molecular weight as those from pancreatic islet extracts. Our study demonstrates the presence of similar differentiation antigens on thyroid C-cells and pancreatic islet cells; this further illustrates common modes of differentiation and specialisation of these embryologically different members of the dispersed neuroendocrine system. The crossreactivity of seven of the monoclonal antibodies investigated with follicular epithelium of the thyroid suggests the existence of common antigenic determinants in different endocrine organs and may partly explain the multiple organ autoimmune response found in patients with polyendocrine diseases. PMID- 2570632 TI - Comparative marker analysis of the ependymocytes of the subcommissural organ in four different mammalian species. AB - The subcommissural organ (SCO), classified as one of the circumventricular organs, is composed mainly of modified ependymal cells, attributable to a glial lineage. Nevertheless, in the rat, these cells do not possess glial markers such as glial fibrillary acidic protein (GFAP), protein S100, or the enzyme glutamine synthetase (GS). They receive a synaptic 5-HT input and show pharmacological properties for uptake of GABA resembling the uptake mechanism of neurons. In this study, we examine the phenotype of several mammalian SCO (cat, mouse, rabbit) and compare them with the corresponding features of the rat SCO. In all these species, the SCO ependymocytes possess vimentin as an intermediate filament, but never express GFAP or neurofilament proteins. They do not contain GS as do glial cells involved in GABA metabolism, and when they contain protein S100 (rabbit, mouse), its rate is low in comparison to classical glial or ependymal cells. Thus, these ependymocytes display characteristics that differentiate them from other types of glial cells (astrocytes, epithelial ependymocytes and tanycytes). Striking interspecies differences in the capacity of SCO-ependymocytes for uptake of GABA might be related to their innervation and suggest a species-dependent plasticity in their function. PMID- 2570634 TI - Covalent regulation of the cardiac sarcoplasmic reticulum calcium pump. Review article. PMID- 2570635 TI - The educational work of the World Federation of Societies of Anaesthesiologists (WFSA). PMID- 2570637 TI - Expression of engrailed proteins in arthropods, annelids, and chordates. AB - engrailed is a homeobox gene that has an important role in Drosophila segmentation. Genes homologous to engrailed have been identified in several other organisms. Here we describe a monoclonal antibody that recognizes a conserved epitope in the homeodomain of engrailed proteins of a number of different arthropods, annelids, and chordates; we use this antibody to isolate the grasshopper engrailed gene. In Drosophila embryos, the antibody reveals engrailed protein in the posterior portion of each segment during segmentation, and in a segmentally reiterated subset of neuronal cells during neurogenesis. Other arthropods, including grasshopper and two crustaceans, have similar patterns of engrailed expression. However, these patterns of expression are not shared by the annelids or chordates we examined. Our results provide the most comprehensive view that has been obtained of how expression patterns of a regulatory gene vary during evolution. On the basis of these patterns, we suggest that engrailed is a gene whose ancestral function was in neurogenesis and whose function was co-opted during the evolution of segmentation in the arthropods, but not in the annelids and chordates. PMID- 2570638 TI - tctex-1: a candidate gene family for a mouse t complex sterility locus. AB - The t complex of the mouse has an important role in male germ cell development and function. Multiple mutations in the t complex interact to alter profoundly the transmission ratio of t complex-bearing sperm or to cause complete sterility or semisterility. We have isolated a multigene family, tctex-1, by screening a testicular cell cDNA library with two reciprocally subtracted testicular cDNA probes. The tctex-1 gene family produces an abundant, virtually germ cell specific transcript that is 8-fold overexpressed in t homozygotes. The aberrant expression of tctex-1 is solely dependent on the t haplotype genes and occurs only in germ cells. The chromosomal location and pattern of expression of tctex-1 make it a candidate for involvement in male sterility. PMID- 2570636 TI - Isolation of a human cyclin cDNA: evidence for cyclin mRNA and protein regulation in the cell cycle and for interaction with p34cdc2. AB - This paper reports the nucleotide and predicted amino acid sequence of a human B type cyclin. The predicted protein sequence shows strong homology to the other known cyclins in the central third of the protein. We show that the level of cyclin mRNA is regulated during the cell cycle, increasing during G2 phase to four time that present in G1. The protein accumulates steadily during G2 to at least 20 times its level in G1 and is abruptly destroyed at mitosis. In G2/M phase, cyclin is associated with p34cdc2, the human homolog of the fission yeast gene cdc2+, and this complex has histone H1 kinase activity. PMID- 2570639 TI - A 60 kd cdc2-associated polypeptide complexes with the E1A proteins in adenovirus infected cells. AB - p60 is a cellular protein that binds to the adenovirus E1A protein complex in virally infected or transformed human cells. In both infected and uninfected cells, p60 was found in a complex with the cdc2 protein kinase. Immune complexes containing p60 and cdc2 display a cell cycle-dependent histone H1 kinase activity that is most active in interphase. The previously described cdc2-p62/cyclin complex also acts as a histone H1 kinase but is maximally active in mitotic metaphase. The shift in the timing of activation of different cdc2-containing complexes suggests that each might play a distinct role in regulation of the cell cycle. PMID- 2570640 TI - The human mRNA that provides the N-terminus of chimeric G6PD encodes GMP reductase. PMID- 2570641 TI - The primary structure of a plasma membrane guanylate cyclase demonstrates diversity within this new receptor family. AB - Atrial natriuretic peptide (ANP) binds directly to a plasma membrane form of guanylate cyclase (GC-A), stimulating the production of the second messenger cyclic GMP. We show that a second guanylate cyclase/receptor (GC-B) exists, with distinctly different specificities for various natriuretic peptides. A cDNA clone encoding GC-B was isolated by low-stringency screening of a rat brain cDNA library using GC-A cDNA as a probe. The deduced amino acid sequence of GC-B is 78% identical with GC-A within the intracellular region, but 43% identical within the extracellular domain. Cyclic GMP concentrations in cells transfected with GC A were half-maximally elevated at 3 nM ANP, 25 nM brain natriuretic peptide (BNP), and 65 nM atriopeptin 1, while 25 microM ANP, 6 microM BNP, and greater than 100 microM atriopeptin 1 were required for half-maximal stimulation of GC-B. The potencies of natriuretic peptides on GC-A and GC-B activity are therefore markedly different; furthermore, despite the specificity of GC-B for BNP, the relatively high BNP concentration required to elicit a response suggests the possible presence of a more potent, unidentified natural ligand. PMID- 2570642 TI - A single glutamic acid residue plays a key role in the transcriptional activation function of lambda repressor. AB - Previous experiments have suggested that negative charge is an important aspect of the activating region of lambda repressor as it is for at least one class of eukaryotic transcriptional activators. Here we randomize amino acids in the activating region of repressor and assay the function of over 100 variants. We find that acidic residues at the four solvent-exposed positions on the surface of an alpha helix (helix 2 in the structure) together comprise a strong activating region. Only one of these acidic residues, however, is critical for activation, and at this position glutamate is strongly preferred to aspartate. At the three remaining positions, certain uncharged residues (different ones at each position) function as well as or better than the acidic residues. Basic residues, however, are highly detrimental to function at all four positions. Our mutagenesis studies also suggest limitations on amino acid substitutions that allow formation of the helix-turn-helix DNA binding motif found in repressor and in many other DNA binding regulatory proteins. PMID- 2570643 TI - Sheep erythrocyte rosetting induces multiple alterations in T lymphocyte function: inhibition of T cell receptor activity and stimulation of T11/CD2. AB - When T lymphocytes from human blood or lymphoid organs are prepared by the sheep red blood cell (SRBC) rosetting procedure, glycoproteins of the SRBC membrane interact intimately with the CD2 (T11) molecule on the T cell surface. We now show that rosette formation has measurable short- and long-term effects upon the T cells. First, for a period of 24-48 hr after rosetting, the signal transducing and activation functions of the T3/Ti T cell antigen receptor complex is paralyzed for anti-T3-induced calcium mobilization, with a concomitant decrease in proliferative response to mitogens or stimulatory anti-T3 antibodies. Calcium mobilization through the alternate pathway of T cell activation, the T11/CD2 SRBC receptor, was also inhibited by rosetting. Second, rosetting appears to confer a partial stimulatory signal through the T11/CD2 pathway. Thus, 72 hr or more after rosetting, there was increased expression of the T11(3) activation epitope, and rosetted T cells were stimulated to proliferate in the presence of anti-T11(3) antibodies alone. These results provide further details on the effects of SRBC-T cell interactions, with important methodological implications. Moreover, they suggest a hitherto unrecognized down-regulatory effect of engaging the CD2 molecule, and provide further evidence that the T cell receptor is functionally interconnected to the CD2 activation pathway. PMID- 2570644 TI - Rheumatoid factor blocks regulatory Fc signals. AB - Immunosuppressed cultures of murine spleen cells, partly deprived of T cells and antigen-stimulated, can be reconstituted to near full activity in their antibody forming cell response with murine rheumatoid factors (RF). The dose of RF required for recovery of 50% of the reconstitutable immune response was 10-100 ng and reconstitution was blocked by intact murine IgG added to the cultures. IgG subclass specificity of RF was demonstrated; RF specific for IgG2a was more potent than RF specific for IgG1 in reconstituting the response. Synergy was observed between RF added at culture initiation and late-acting B-cell differentiation factors. The greater the degree of T-cell deprivation, the more stringent the conditions needed for reconstitution. Suitable conditions for reconstitution with profound T-cell depletion included the limited reconstitution by specific RF, the synergistic action of RF with late-acting T-cell-replacing supernatants, and multiple additions of a number of RFs to the cultures on Days 0, 1, and 2. RF was also shown to block Fc-dependent immunosuppression by added antigen-antibody complexes. These results are interpreted as favoring the hypothesis put forward previously that the normal production of RF acts to reduce T-cell dependency by preventing negative Fc signal transmission by immune complexes on the B-cell surface. Abnormal production of RF may be a primary destabilizer of the immune responses leading to autoimmunity. PMID- 2570645 TI - OKT4 monoclonal antibody-induced activation of an autoreactive T-cell clone. AB - We report the presence of a CD4-mediated T-cell activation pathway on an autoreactive CD3+WT31+CD4+CD8- T-cell clone, designated 2F9, isolated from the peripheral blood of a patient with ovarian adenocarcinoma. The OKT4 mab modulated the CD4 antigen independently of the CD3 antigen or the alpha beta T-cell receptor. OKT4 mab immobilized on plastic or soluble OKT4 mab in the presence of feeder PBMC induced proliferation and IL-2 production by cells of the 2F9 clone. Mixtures of the OKT4 mab and the OKT3 or anti-WT31 mabs induced additive proliferative responses and IL-2 production. The OKT4 mab synergized with recombinant IL-2 in inducing proliferative responses. These results suggest the presence of activation pathway on 2F9 cells. PMID- 2570646 TI - CD4+ cytolytic T cell clones restricted to HLA class II, DR beta I, and DR beta III chains. AB - We have investigated the functional polymorphism of HLA class II antigens using CD4+ CTL clones. Seven CD4+ CTL clones were isolated from a healthy donor (HLA A2 A24; B8 B27; DRw17 DRw52a) by repeated stimulation with irradiated autologous EBV transformed B cell lines (EBV-B). According to the HLA restriction specificity we divided CD4+ CTL clones into three subgroups: (i) DRw17-restricted CD4+ CTL clones; (ii) DRw52a-restricted CD4+ CTL clones; and (iii) the CD4+ CTL clones, of which the restriction specificity could not be assigned to products of a single HLA locus. Interestingly, DRw17-restricted CD4+ CTL clones distinguished between DRw17 and DRw18. Similarly, DRw52a-restricted CD4+ CTL clones distinguished between DRw52a, w52b, and w52c. There are four amino acids which differ between DRw17 and DRw18, whereas five differ between DRw52a and the other two alleles (DRw52b and DRw52c). The recent elucidation of the crystal structure of a human class I MHC molecule has identified the probable peptide binding site to be a cleft on the outer surface of the molecule, between two alpha-helices. On the basis of the theoretical model for HLA class II molecules, amino acid positions 26 and 28 (DRw17 vs DRw18) and amino acid positions 26, 28, and 74 (DRw52a vs the other two alleles) lie within the "cleft." We propose that amino acid positions 26 and 28 are very important sites with regard to the recognition of antigen-MHC complex by the TCR. PMID- 2570647 TI - Formation of a direct mutagen, diazo-N-nitrosoetilefrin, by interaction of etilefrin with nitrite. AB - Reaction of an antihypotensive drug, etilefrin [alpha-[(ethylamino)methyl]-m hydroxybenzyl alcohol], with nitrite under mildly acidic conditions produced N nitrosoetilefrin [alpha-[(N-nitrosoethylamino)methyl]-m-hydroxybenzyl alcohol] (a mixture of syn and anti forms) (Iab) and diazo-N-nitrosoetilefrin [1-(4-diazo-3 oxo-1,5-cyclohexadienyl-2-(N-nitrosoethylamino )ethanol] (a mixture of syn and anti forms) (IIab). Treatment of etilefrin with an equivalent amount of nitrite at pH 3 and 37 degrees C for 4 h gave Iab (yield, 30%) and IIab (yield, 5%). Treatment of etilefrin with 4 eq of nitrite under the same conditions gave Iab (23%) and IIab (53%). Compounds Iab and IIab were each composed of two isomers due to the configuration of the N-nitroso group. While compound Iab was not mutagenic, compound IIab showed mutagenicity to Salmonella typhimurium TA98 and TA100 strains without metabolic activation. Specific mutagenic activity of IIab was 300 his+ revertant colonies for both TA98 and TA100 strains with a dose of 0.1 mumol. Addition of a microsomal activation system little affected the activity. It is noteworthy that this orally administered drug can produce a direct-acting mutagen by reaction with nitrite, which is present in the digestive tract. PMID- 2570649 TI - Developmental regulation of tyrosine hydroxylase in the mediobasal hypothalamus. AB - In the adult rodent the mediobasal hypothalamus (MBH) interacts extensively with the pituitary gland to regulate a variety of endocrine functions. The dopaminergic (DA) neurons of the mature MBH are influenced by numerous transmitters and hormones, however, little is known about developmental regulation of this system. Ontogeny of DA neurons was examined in vivo and in explant culture by monitoring tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis. The influence of the depolarizing agent, veratridine, was examined to determine whether development of TH in the MBH is regulated by depolarizing signals as in other CA neurons. Veratridine elicited a significant increase in TH activity in cultures of MBH. Adult MBH neurons are influenced by hormones such as estradiol. We investigated the possible role of estradiol in regulating the ontogeny of MBH DA neurons in culture. We developed a steroid-depleted culture medium to rigorously define the effects of steroids on the developing system. This enabled us to determine that estradiol does not appear to influence TH during embryonic development, though estrogen receptors are present at this stage. These results were confirmed in vivo by injecting neonates with moxestrol, a synthetic estrogen which is not sequestered by alpha fetoprotein. This treatment did not elicit any change in TH. Our observations suggest that although estrogen regulates TH in the adult MBH, this hormone does not play a role in developmental regulation of TH in this brain region. In contrast, however, depolarizing signals appear to be a widespread mechanism for regulation of TH in numerous neuronal populations. PMID- 2570648 TI - [A histochemical study of diethylnitrosamine-induced altered liver cells in rats]. AB - The histochemical changes of gamma-glutamyltransferase (gamma-GT), adenosine triphosphatase (ATPase), glucose-6-phosphatase (G-6-Pase) and ornithine carbamyltransferase (OCT) were studied in diethylnitrosamine (DEN) -induced and enzyme-altered liver cell lesions (Solt-Farber model) in rats. The number of altered liver cell foci tended to decrease after ceasation of 2 acetylaminofluorene (2-AAF); nevertheless, the number and size of the nodules increased rapidly within 20 weeks. The histochemical changes of most of the altered liver cell foci were focused on one or two kinds of enzyme activity (mostly gamma-GT and ATPase); while most of the nodules presented 3 or 4 kinds of histochemical changes, including OCT and G-6-Pase. It is concluded that some of those altered nodules of multi-enzyme changes might develop continuously to become tumors. PMID- 2570650 TI - Enzymatic imbalance in peripheral blood mononuclear cells isolated from individuals with anti-HIV antibodies. AB - Plasma membrane-bound 5'-nucleotidase (5'-NT), gamma-glutamyltransferase (gamma GT) and soluble deoxynucleotidyltransferase (TdT) were studied in peripheral blood cells (PBMN) of 35 individuals, 26 male and 9 female, with circulating anti HIV antibodies. Twenty-six were drug abusers, 2 were drug abusers and homosexuals and 4 were homosexuals. Three did not fall into any risk group. The surface immunologic phenotype of cells stained with the fluorescent monoclonal antibodies Leu 5, Leu 3, Leu 2, Leu 12, Leu M3, Leu M1, anti-CALLA and anti-HLA-DR was delineated by flow cytometry. While the gamma-GT activity did not change, the lymphocyte 5'-NT activity was significantly less than normal in anti-HIV positive individuals and in anti-HIV negative drug abusers. TdT activity was detectable in 14 anti-HIV positive patients (40%), who did not have clinical AIDS. Of 8 patients with AIDS, 3 had a low level of TdT activity but 5 had cells completely devoid of TdT and 5'-NT activity. 5'-nucleotidase activity and the frequency of Leu 2 suppressor antigen bearing cells were the only independent variables that correlated with AIDS incidence. PMID- 2570651 TI - DNA fingerprinting. AB - Hypervariable tandem-repetitive minisatellite regions of human DNA can be used to generate individual-specific DNA fingerprints. Validation studies have demonstrated the reliability of the analysis, the mode of inheritance of the minisatellites, and the unparalleled degree of individual specificity. The uses of hypervariable probes in forensic biology, paternity testing, and the resolution of a wide range of problems in genetics, molecular biology, population biology, and medicine are illustrated. PMID- 2570652 TI - Use of polymerase chain reaction for diagnosis of inherited disorders. AB - The polymerase chain reaction (PCR) is a rapid method for generating a 10(6)- to 10(7)-fold increase in the number of copies of a discrete DNA or RNA sequence. The technique is being used for rapid prenatal diagnosis and carrier testing of several inherited disorders. After PCR, mutations producing single-gene disorders can be detected by several different methods, including endonuclease digestion and gel electrophoresis (applicable when a mutation affects an endonuclease recognition site), gel electrophoresis (used for detection of deletions), and hybridization to an oligonucleotide probe specific for a mutation. Less often, gene sequencing of a PCR product is used to rapidly identify a mutation. In addition, the PCR technique can be applied to polymorphism analysis to provide diagnosis by linkage analysis. In other areas, PCR is being used to detect and characterize microbial pathogens and to characterize mutations associated with carcinogenesis. The PCR method is useful in situations in which the amount of DNA sample is limited, such as in forensics and prenatal testing, or in which the quality of the DNA sample is poor. PMID- 2570654 TI - Hereditary localized pruritus in affected members of a kindred with multiple endocrine neoplasia type 2A (Sipple's syndrome). AB - We describe a kindred with medullary thyroid carcinoma and phaeochromocytoma (MEN 2A or Sipple's syndrome) in which a cutaneous manifestation is only present in affected members. These members felt an intense itching in an area 5-10 cm in length and passing through the scapular region where, after long-term scratching, the skin appears hyperkeratotic and pigmented. Cutaneous biopsies proved negative for dermis nerve abnormality. This restricted itching strongly suggests dominant transmitted hereditary localized pruritus which was described many years ago in a family which was apparently free from inherited diseases. In the examined kindred this skin peculiarity was said to have appeared before the patients reached 10 years of age and, therefore, prior to the biochemical manifestation of the thyroid tumour. Three children of the last generation, aged 4 to 11 years, all of whom presented this cutaneous manifestation, were considered unaffected because of normal basal and stimulated calcitonin, but thyroid C-cell hyperplasia was recently proved in the eldest. In this MEN 2A kindred the presence of such a characteristic hereditary itch in affected members may be considered as a phenotypic marker giving advance warning of medullary thyroid carcinoma. PMID- 2570653 TI - TSH binding-inhibiting antibodies in hyperthyroidism: relationship to clinical signs and hormone levels. AB - TSH binding-inhibiting antibody (TBIAb) activity was measured in 809 European patients with different forms of hyperthyroidism. Distribution of these TBIAb was skewed, with a peak in the range of normal controls, and an ill defined, not clearly separated peak at higher levels of TSH displacement. There was no unequivocal separation of two possible subgroups of hyperthyroidism (immunogenic and non-immunogenic). TBIAb distributions of patients with and without endocrine ophthalmopathy (EO) overlapped considerably. Although patients with Graves' disease, arbitrarily defined by the presence of endocrine ophthalmopathy or diffuse nuclide uptake by thyroid scanning, had mostly elevated TBIAb activity, 24.3% had values within the range of normal controls (mean + 2SD). Patients with diffuse thyroid uptake had significantly higher TBIAb levels than patients with nodular scan findings. In Graves' disease, TBIAb activity was positively correlated with the severity of endocrine ophthalmopathy, the size of the thyroid, and the serum levels of total and free triiodothyronine. There was no influence of age, sex, pretreatment, or regional iodine supply. These results suggest (1) that the clinical manifestations of Graves' disease are statistically related to TBIAb activity and (2) that separation of immunogenic and non immunogenic forms of hyperthyroidism by means of TBIAb determination is unsatisfactory. The almost continuous distribution of TBIAb points to insufficient sensitivity of the present technique and raises doubts as to whether TBIAb values can be reliably classified as 'positive' or 'negative'. PMID- 2570655 TI - Enterohepatic circulation of opioid drugs. Is it clinically relevant in the treatment of cancer patients? PMID- 2570656 TI - Healing and recurrence of active duodenal ulcer with nizatidine. AB - Nizatidine, a new H2-receptor antagonist for treatment of duodenal ulcer disease, was evaluated in a unique two-phase, placebo-controlled, randomized, double blind, multicenter clinical trial. Patients received either 150 mg nizatidine twice daily or placebo for 4 weeks (phase I). If ulcer healing did not occur during phase I, patients were randomly reallocated to receive either 150 mg nizatidine twice daily or placebo for an additional 4 weeks (phase II). Patients with a healed ulcer continued on the same therapy. All patients were endoscoped at week 8. Healing rates at week 2 were 93 of 265 (35%) nizatidine-treated patients and 55 of 260 (21%) placebo-treated patients (p less than 0.001); at week 4, healing rates were 198 of 259 (76%) nizatidine-treated patients and 95 of 243 (39%) placebo-treated patients (p less than 0.001). In phase II, ulcer healing occurred in 46 of 86 (53%) nizatidine-treated patients and in 23 of 90 (26%) placebo-treated patients (p = 0.002). In patients who had a healed ulcer at previous endoscopies, 18 of 178 (10%) nizatidine-treated patients and 10 of 81 (12%) placebo-treated patients had a recurrence of duodenal ulcer. Smokers who had histories of previous ulcers were more likely to have an early recurrence. PMID- 2570658 TI - The Harderian gland. PMID- 2570657 TI - Plasma calcium difference between man and vertebrates. AB - 1. Literature data about the plasma content of total calcium, ionized calcium and inorganic phosphate in healthy animals and man of different age and sex were collected. 2. It was established that under normal conditions ionized calcium is about 45% of total calcium. 3. The degree of saturation of these blood samples with respect to octocalcium phosphate OCP was calculated. 4. In young animals and man the blood plasma has a higher degree of saturation than in adult animals and man. 5. The blood plasma of healthy animals is supersaturated with respect to OCP during their whole life. 6. However, the blood plasma of healthy human adults is slightly undersaturated with respect to OCP. PMID- 2570659 TI - Ultrastructural and biochemical evidence of glycogen in the developing lung of the chick embryo: possible contribution to surfactant. AB - 1. Glycogen was identified ultrastructurally in undifferentiated type-II cells of the lung of the day 16 chick embryo. 2. By 4 days after hatching, glycogen in type-II cells could not be observed, although lungs were actively secreting surfactant. 3. Biochemical measurements of pulmonary glycogen revealed a depletion during days 14-20 of incubation, corroborating ultrastructural data. 4. Using lung slices, 14C-glucose was incorporated in vitro into pulmonary surfactant phospholipids at a high rate in day 14 embryos, and a significantly lower rate on day 19. 5. Hypophysectomy resulted in sub-normal initial accumulation of pulmonary glycogen on day 14 of development, but did not alter the depletion pattern after day 16. 6. Thus, glycogen stores may contribute to avian embryonic pulmonary surfactant, and accumulation of early stores may be under hormonal control. PMID- 2570660 TI - Oxygen consumption of brown noddy (Anous stolidus) embryos in a quasiequilibrium state at lowered ambient temperatures. AB - 1. The oxygen consumption (MO2) of the semi-precocial Brown Noddy embryos at different stages of development was measured at 36 degrees C and again after 5-hr exposure to lowered ambient temperatures (30 and 32 degrees C). 2. The MO2 measured in a quasiequilibrium state was equal to the value predicted by a temperature coefficient of 2. 3. In contrast to precocial chickens, the semi precocial Noddy had no apparent metabolic response to cooling before hatching. PMID- 2570661 TI - A biphasic excitability change in hindlimb motoneurons evoked by stimulation of the nucleus raphes magnus in the cat. AB - 1. The influence of electrical stimulation of the nucleus raphes magnus (RM) on spinal segmental systems were examined. 2. RM stimulation produced an initial increase and a subsequent suppression of the amplitude of both fiextor and extensor lumbar monosynaptic reflex potentials (MSRs). 3. Intracellular recordings were made from alpha-motoneurons of the common peroneal nerve (flexor) and the tibial nerve (extensor). RM stimulation evoked postsynaptic potentials with a time course similar to that of MSR facilitation. 4. RM stimulation inhibited the aggregate excitatory synaptic potential (EPSP) evoked by stimulation of group I afferent fibers without apparent changes in the motoneuronal membrane potential. 5. These data suggest that the RM-evoked biphasic effect on MSR consists of early facilitation due to EPSP, and late inhibition possibly due to presynaptic inhibition of group I afferent fibers. PMID- 2570662 TI - Stimulation of HCO3- excretion in frog skin by urinary and adrenal cortical extracts. AB - 1. There is a substance extractable from human urine which stimulates frog skin to excrete bicarbonate. The response to this extract is delayed, taking 4-9 hr, depending upon the dosage. 2. The amount of this substance varies with the bicarbonate intake of the urine donor. 3. This is a reasonable dose-response curve to the extract. 4. A substance with the same solubility characteristics, and which stimulates bicarbonate excretion with a similar time response is present in adrenal cortical extracts. PMID- 2570663 TI - Effect of heat stress on plasma levels of arginine vasotocin and mesotocin in domestic fowl (Gallus domesticus). AB - 1. The effect of an acute heat stress on plasma arginine vasotocin (AVT) and mesotocin (MT) levels were determined in non-heat-acclimated domestic fowl. 2. Heat increased (P less than 0.05) AVT and decreased MT (P less than 0.05) levels but had no affect on plasma osmolality. Changes in MT were correlated with body temperature (-0.40) and blood pCO2 (0.32). However, changes in AVT were not correlated with any physiological variable, including MT. 3. During thermoneutral recovery, with the exception of hematocrit which remained lower (P less than 0.05), all monitored variables returned to values which were not different (P greater than 0.05) from those observed prior to heat stress. 4. These results indicate that in non-heat-acclimated birds, acute heat stress elevated plasma AVT and suppressed the release of MT and that changes in these hormones occurred independent of any change in plasma osmolality. PMID- 2570664 TI - Effect of hemorrhage on hepatic tissue blood flow in the domestic fowl (Gallus domesticus). AB - 1. Thermal Pulse Decay (TPD) methodology was used to monitor hepatic tissue blood flow (hepatic perfusion) in anesthetized birds prior to and during hemorrhagic hypotension. 2. Hemorrhage was accomplished by periodic removal of blood through a carotid cannula. Reducing the estimated blood volume (BV) from 100 to less than 50% decreased hepatic perfusion from 4.36 +/- 0.7 to 1.88 +/- 0.7 ml/min/g. 3. Changes in hepatic perfusion during the experiment were related to mean arterial blood pressure (MABP) by the following linear regression equation: hepatic perfusion = -1.79 +/- 0.0807x (r2 = 0.57). PMID- 2570665 TI - Skinned muscle fibers of Aplysia: potentiation of contraction by "background" calcium and lack of effect of cyclic AMP. AB - 1. Aplysia buccal muscle E1 can be skinned with saponin in a low ionic strength medium. Pulses of calcium, which were ineffective at causing contraction in intact fibers, elicited contraction in skinned fibers. 2. Tension in skinned fibers increased at [Ca2+] greater than 10(-7) M and was maximal at 6 x 10(-7) M. 10(-5) M [Ca2+] caused irreversible damage to the fibers. 3. Fibers did not exhibit "catch", i.e. they relaxed quickly upon removal of calcium. 4. Optimal pH for tension was 7.0. 5. Contractile responses to calcium pulses were increased by raising "background" [Ca2+] to 10(-7) M. 6. Cyclic AMP (10(-4) and 10(-3) M) had no effect on tension. PMID- 2570666 TI - Effects of photoperiod, melatonin implants and castration on molting and on plasma thyroxine, testosterone and prolactin levels in the European badger (Meles meles). AB - 1. The seasonal molt, which lasts six months in the badger, begins in mid-July and ends at the beginning of winter. It occurs under natural long-day conditions, following the seasonal drop in plasma testosterone levels, concomitant with high levels of thyroxine and prolactin. 2. To examine the role of the different factors involved (day length, prolactin, thyroxine, testosterone), different groups of badgers, divided into subgroups of castrated or intact animals, were subjected to the influence of long days (20L: 4D), short days (4L:20D) or the effect of subcutaneous melatonin implants. 3. In all cases, castration resulted in a significantly earlier onset of molting 1-3 months, depending on the group, regardless of the experimental conditions (20L:4D, 4L:20D, melatonin). 4. However, molting started earliest in animals subjected to long days, irrespective of whether they were castrated or intact. 5. In the melatonin-implanted badgers, molting started either early (castrated animals), or late or not at all (intact animals). 6. Lastly, in castrated badgers subjected to experimental photoperiods (short days or long days) or melatonin implants, the period of molting was shortened from 6 months (intact outdoor animals) to 4 months. 7. The advance in shedding was always related to an early drop in testosterone (or an absence of testosterone in the castrated animals) and to a higher or earlier increase in thyroxine levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570667 TI - Influence of egg production on zinc, copper and iron metabolism in the turkey hen (Meleagris gallopavo). AB - 1. Serum concentrations of zinc, copper, iron, calcium and protein increased significantly with the onset of egg production in turkey hens. 2. Hepatic concentrations of zinc, copper and iron declined significantly with the onset of egg production, whereas the total hepatic content of zinc and copper increased significantly. 3. When plasma from laying and nonlaying hens was subjected to gel permeation column chromatography, a high molecular weight protein fraction was detected in laying but not in nonlaying plasma which bound significant amounts of zinc, copper and iron and which coeluted with purified vitellogenin. 4. Vitellogenin purified from the plasma of laying hens is a metalloprotein which upon analysis was found to contain 0.54, 0.09 and 0.72 micrograms of zinc, copper and iron, respectively, per mg of protein. PMID- 2570668 TI - Proton efflux from rat intestinal basal-lateral membrane vesicles is stimulated by ATP and Na+. AB - 1. ATP-stimulated 22Na uptake and 14C-methylamine efflux were studied in inside out rat intestinal basal-lateral membrane vesicles (BLMV). 2. Uptake of 22Na by basal-lateral membrane vesicles was stimulated by addition of ATP and by an acidic vesicle interior. 3. Efflux of 14C-methylamine was stimulated by ATP and Na+. 4. 14C-methylamine efflux was not influenced by vanadate or amiloride by themselves but was inhibited by the presence of both agents. 5. These data are consistent with a basal-lateral proton translocation mechanism which may be responsible for alkalinization of the lateral intercellular space and implicates the Na+-pump in this mechanism. PMID- 2570669 TI - Chemical specificity of short-chain fatty acid-induced electrogenic secretory response in the rat colonic mucosa. AB - 1. Luminal additions of normal short-chain fatty acids (SCFA: 3-6 carbons) increased transmural potential difference (PD: lumen negative) across rat everted colon in a dose-dependent manner. 2. The PD response to SCFA adapted rapidly and was not evoked by further cumulative addition of the same acid (self-adaptation) or different acids (cross-adaptation). 3. A broad range of SCFA derivatives were tested as potential stimulators of the PD response and cross-adaptation with propionate: a single carboxyl group and 3-6 carbon-chain were an absolute requirement for recognition. 4. These results suggest that SCFA-reception mechanism of the rat colonic mucosa has some sort of chemical specificity for stimulating electrogenic secretory response. PMID- 2570671 TI - The morphology and electrophysiology of the neurones of the paired pedal ganglia of Lymnaea stagnalis (L.). AB - 1. A morphological and electrophysiological map of the identifiable neurones and neuronal clusters of the paired pedal ganglia has been prepared. 2. Neuronal morphology was investigated using the fluorescent dye, Lucifer Yellow CH, whilst electrophysiological properties were studied using conventional intracellular recording techniques and the phase plane technique. 3. The paired pedal ganglia are largely symmetrical and giant neurones usually have contralateral homologues. 4. Neuronal clusters are also paired, but minor asymmetries, both of identifiable neurones and neuronal clusters have been found to exist. 5. These asymmetries are thought to be related to asymmetries of body form. 6. Most of the individually identifiable neurones possess obligatory axon branches which are invariant from one preparation to the next, but variant branches also occur. 7. Within the neuronal clusters, morphology appears to be more variable. 8. Individually identifiable neurones and neuronal clusters were characterized electrophysiologically according to the criteria of action potential shape, spontaneous activity pattern, electrical coupling and common synaptic inputs. 9. Homologous pairs of neurones usually have similar electrophysiological properties, as do those within clusters. 10. A number of wide-acting synaptic inputs have been identified on neurones of the pedal, buccal, visceral and parietal ganglia. PMID- 2570670 TI - Changes of basal and steroidal precursor-stimulated testosterone secretion in isolated Mongolian gerbil and guinea pig testes after a single episode of heating. AB - 1. In the absence of steroidal precursors, testosterone secretion by Mongolian gerbil testes incubated at 37 degrees C was 340 ng/g tissue/4 hr. Addition of 1 microgram progesterone or DHEA drastically stimulated testosterone secretion by testes incubated at 37 degrees C (progesterone: 3281 ng/g tissue/4 hr, DHEA: 4654 ng/g tissue/4 hr). 2. While neither basal nor DHEA-stimulated production of testosterone was significantly affected by a single episode of heating (43-44 C for 30 min), progesterone-stimulated testosterone secretion markedly decreased during the 4-hr incubation period. 3. In contrast, in isolated testes of adult guinea pigs, a single episode of heating (44 degrees C for 30 min) resulted in a drastic reduction of basal and precursor-stimulated testosterone production during the 4-hr incubation period. 4. From these data it appears that enzymatic activities in the testes of the two species do not have their maxima at the same temperature, but rather in each case at, or close to, the temperature prevailing in the scrotal testis. PMID- 2570673 TI - The biochemical ecology of Biomphalaria glabrata, a snail host of Schistosoma mansoni: short chain carboxylic and amino acids as phagostimulants. AB - 1. A buccal mass olfactometer was used to investigate the responses of the fresh water pulmonate snail Biomphalaria glabrata to carboxylic and amino acids. 2. The snails proved very discriminating as only 6 (14.6%) of the 41 chemical species tested were effective as phagostimulants. These are ranked as follows in order of potency:- butanoate greater than propanoate greater than D-malate greater than 2 hydroxybutanoate = L-tartrate = L-aspartate. 3. The structure-activity relationships of the active compounds, and their significance to the ecology and control of the snails are discussed. PMID- 2570672 TI - Relationships between a neuronal buccal population and the peribuccal regions in Aplysia: an electrophysiological study. AB - 1. The relationships between Aplysia buccal neurons projecting the cerebral ganglion (L cells) and peribuccal regions were studied by electrophysiological techniques. 2. Stimulation of the cerebral upper labial (UL) and anterior tentacular (AT) nerves produced excitatory postsynaptic potentials in L cells. 3. Sixteen cells out of 24 were found possess an axonal branch in the labial branch of the AT nerve, 1 out of 8 in the UL nerve. 4. These axonal branches did not show any direct motor or sensory function in "reduced" preparations. 5. A modulatory function for the axonal projections and a sensory role for the synaptic relationships are hypothesized. PMID- 2570674 TI - Effects of moderate intensity treadmill exercise on triglyceride production in the laying fowl. AB - 1. Sustained aerobic exercise in domestic fowl has previously been shown to enhance lipid utilization by the skeletal muscles. The present study examined the possibility that such increased lipid oxidation in the laying female might lower the production of plasma triglycerides destined for transfer to the developing oocytes. 2. Following an intravenous injection of 25 muCi of 14C-labelled glycerol trioleate, the experimental birds performed 90 min of treadmill exercise at a work intensity approximately equivalent to 2.5 times the resting metabolic rate. 3. There was no evidence of either glycogen or triglyceride depletion in either the leg muscles or the viscera of the exercised birds. The specific activity of triglyceride TG-SRA found in the tissues was also the same in control and experimental birds. The time-course of the changes in plasma TG-SRA throughout the experimental period gave no indication that TG production had been affected by exercise. 4. It is concluded that the increased energy substrate demand arising from moderate-intensity, aerobic exercise is almost fully met by the liberation of fatty acids from adipose tissue TG stores, and has minimal impact on the hepatic manufacture of egg lipids. PMID- 2570675 TI - The mouse Cebp gene encoding a DNA-binding protein is polymorphic and is located on chromosome 7. AB - The mouse gene Cebp, encoding the DNA-binding protein C/EBP, has been localized to the proximal region of chromosome 7 by determining the strain distribution patterns of a restriction fragment length polymorphism among the BXD and AKXL recombinant inbred mouse lines. PMID- 2570677 TI - Molecular definition of de novo and genetically transmitted WAGR-associated rearrangements of 11p13. AB - We describe a family in whom the phenotypically normal father carries a balanced insertional translocation, ins(14;11)(q23;p12p14). This individual fathered three mentally retarded children, two with a del(11)(p13) and one with a dup(11)(p13). Two other cases of a de novo del(11)(p13) are also described. All four del(11)(p13) cases presented with WAGR, a complex syndrome associated with a predisposition to Wilms' tumor (WT), aniridia (A), genitourinary abnormalities (G), and mental retardation (R). Using an approach combining karyotype analysis, determination of the gene copy number, and RFLP studies employing five 11p13 DNA markers, we were able to define the chromosomal rearrangement involved in each case. Analysis of these WAGR deletions provides further subdivision of band p13 on chromosome 11. PMID- 2570676 TI - Exclusion analysis of Charcot-Marie-Tooth neuropathy (CMT1) with chromosome 1p markers. AB - We previously described a large five-generation family with autosomal dominant inheritance of hereditary motor and sensory neuropathy type I, or Charcot-Marie Tooth disease (CMT1). The genetic defect in this family was not linked to the Duffy blood group. We investigated the possibility of a disease locus on the short arm of chromosome 1 using 12 anonymous DNA markers. Two markers, D1S2 and D1S22, showed positive linkage, suggesting the existence of a CMT1 locus on 1p. D1S2 and D1S22 are clustered in the 1p31----p22 region. However, multipoint linkage analysis, including additional DNA markers from this chromosome region, excluded a possible CMT1 locus in this part of chromosome 1. PMID- 2570679 TI - The duration of viremia patients with epidemic hemorrhagic fever. AB - The duration of viremia in patients with epidemic hemorrhagic fever (EHF) was studied using immunofluorescence technique and cell culture assays. The duration of virus in plasma of EHF patients was about 1 week, which was concomitant with the febrile phase of the disease. Comparatively, the separation rate of viruses in peripheral blood mononuclear cells (PBMC) was greater (1.8 times) than in plasma, while the detected peak of EHF (in 4-7 days after onset of the disease) was found to be 2 or 3 days less than that of plasma, thus the detectable viremia was prolonged until the 8th to 11th day of the disease. High titre EHF antibody of IgG, TgM was unable to neutralize the virus in human blood. The results revealed that a blood-cell-associated prolonged viremia is one of the characteristics of EHF infection. PMID- 2570678 TI - Chromosomal assignment of 14 genomic probes for highly polymorphic loci. AB - Over 500 probes revealing restriction fragment length polymorphisms (RFLPs) have been isolated by Schumm et al. (1988). We describe here the chromosomal assignment of 14 of the most highly polymorphic markers in that set of probes, with polymorphism information content values of up to 0.98. The probes were mapped using a panel of human x rodent somatic cell hybrids and were found to be distributed among nine different autosomes. Chromosome localization of such highly polymorphic markers has been an important step in the construction of the human genetic map, as a large number of RFLP probes has now been localized by genetic linkage studies to these loci. PMID- 2570680 TI - Anti-shock effects of synthetic effective compositions of fructus aurantii immaturus. Experimental study and clinical observation. AB - Satisfactory results have been obtained in treating infective shock with injection of natural Fructus Aurantii immaturus (nat-FAI). The results of animal experiments and clinical observations on the anti-shock effects of the synthetic effective compositions of Fructus Aurantii immaturus (syn-FAI) are reported. The cardiac output increased from 0.53 to 0.87 L/min (P less than 0.01), and the cardiac index increased from 0.99 to 1.63 L/m2/min (P less than 0.01) in the endotoxic shock dogs after the treatment with syn-FAI. At the same time the blood stream in bulbar conjunctiva became accelerated and the dilated microvessels began to get smaller in most dogs. Of fifty children with infective shock treated with syn-FAI, forty-eight showed curative effects, with a total effective rate of 96%. The anti-shock effective compositions in FAI have been proved to be synephrine and N-methyltyrosamine. Moreover, syn-FAI has shown a more stable property, less side-effects and better clinical results than nat-FAI. PMID- 2570681 TI - Heterochromatic regions in different Drosophila melanogaster stocks contain similar arrangements of moderate repeats with inserted copia-like elements (MDG1). AB - Seven out of twenty 30-50 kb genome fragments with an MDG1 copia-like element cloned in cosmids were found to carry homologous sequences which belong to a new family of non-mobile heterochromatic moderate repeats (the HMR family). These repeats along with the MDG1 copies inserted in them are under-replicated in polytene chromosomes. Such repeats may also be located in the intercalary heterochromatin site 12E of the X chromosome. Chromosomal heterochromatic regions are enriched with one of the two main genomic variants of MDG1, MDG1het, identifiable by EcoRI restriction. From Southern DNA blot analysis the number of MDG1het copies and their sites within the heterochromatin are invariant in all the stocks examined, while there is not a single MDG1 site along the polytene chromosomes shared by all the stocks in question. PMID- 2570682 TI - Photobiology of natural populations of zooxanthellae from the sea anemone Aiptasia pallida: assessment of the host's role in protection against ultraviolet radiation. AB - Natural populations of the sea anemone Aiptasia pallida containing endosymbiotic dinoflagellates were acclimated to different irradiance regimes, with and without ultraviolet radiation (UV). They showed a compensatory response in the amount of chlorophyll and the activities of enzymes responsible for detoxifying active species of oxygen that are produced by the interaction between visible or ultraviolet radiation and photosynthetically produced oxygen. Protection from active species of oxygen is essential to prevent the photooxidation of chlorophyll and the concomitant loss of productivity. Bulk analyses of chlorophyll showed differences between the populations exposed to varying irradiance regimes, but revealed no significant independent effect of UV. However, analysis by flow cytometry of the individual cells from treated populations did detect statistically significant differences in cell size and the amount of chlorophyll fluorescence per cell, which could be attributed to treatment with ultraviolet radiation. With flow cytometry we are able to detect the population variability that is undetectable by bulk measurements which is important in assessing the effects of environmental parameters in both symbiotic and free-living microalgae. Research using simultaneous measurements by flow cytometry could add considerable insight into the population dynamics of both zooxanthellae and host cells. PMID- 2570684 TI - [Pre-neoplastic sign in human esophagus]. AB - Gamma-glutamyl Transpeptidase (gamma-GT) activity of abrasive balloon cytocollections from esophagus of a population aged above 35 in high risk area was studied. Gamma-GT positive rate of the 931 subjects studied was increasingly elevated with the severity of histopathologic changes as follows: normal group 10.3% (43/419), hyperplasia group 21.9% (46/210), dysplasia group 43.9% (125/285), pre-carcinoma group 100% (4/4), squamous carcinoma group 77% (10/13). All or most of the gamma-GT positive cells of each group were normal esophageal squamous epithelium (NESE). They may be initial transformed cells similar to discrete gamma-GT rich cell population without morphological changes observed during very early stages of chemical carcinogenesis. In addition, gamma-GT positive hyperplastic cells and dysplasia grade I cells were also observed in some cases, but dysplasia grade II cells with gamma-GT activity were only shown in a few cases. 1-1.5 years after the mass screening, all the gamma-GT positive cases and gamma-GT negative dysplasia cases were planned to be followed. However, only 277 cases were followed and observed. Histopathologic changes developed rapidly in fifteen cases and fourteen of them were gamma-GT positive cases 1-1.5 years before. Thus, the development of gamma-GT activity may show the trend of histopathologic progression. Gamma-GT on NESE, hyperplastic cells and dysplasia grade I cells showed two fates--progression or fade, whereas the fade rate was reversed with the severity of histopathologic changes. Thus, these changes may be the sign of doubtful population to different extents.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570685 TI - [In vitro formation of multimers of specific DNA-protein complexes with nuclear factor I (NF I)]. PMID- 2570683 TI - Characterization of autonomic receptors in neonatal urinary tract smooth muscle. AB - In the 1-day-old rabbit there are a large number of alpha-2- (labeled with [3H]yohimbine) than alpha-1- (labeled with [3H]prazosin) adrenergic receptors in the various parts of urinary tract smooth muscle, but the alpha-1 receptors are functionally more responsive to agonists than are the alpha-2 receptors. Unlike the findings in the adult rabbit, the neonatal bladder dome is functionally more sensitive to alpha-adrenergic agonists than the neonatal bladder base. The rank order of beta-adrenergic receptor densities (labeled with [3H]dihydroalprenolol) in various urinary tract smooth muscles correlates with the magnitude of the relaxant response to isoproterenol. There is a high number of functional muscarinic receptors (labeled with [3H]quinuclidinyl benzylate) in the neonatal urinary tract. The data demonstrate the existence of functional autonomic receptors in neonatal urinary tract smooth muscle. PMID- 2570686 TI - Evidence of drug interaction between cyclosporin A and H2-receptor antagonists (cimetidine, ranitidine and famotidine) in Sprague-Dawley rats. AB - H2-receptor antagonists, such as cimetidine (C), ranitidine (R) and famotidine (F) seem to be effective in the prevention and treatment of stress ulcer in transplant recipients receiving cyclosporin A (CyA). The aim of this study was to detect the possible synergistic nephro- and hepato-toxicity of these drugs, assaying the serum creatinine (SC), ALT, AST levels, and the histological features of 45 young male Sprague-Dawley rats, divided into nine groups of five rats each. After 10 days of treatment the results showed: (i) serum CyA levels were increased in the group receiving daily CyA (5 mg/kg) + R(5 mg/kg) (2430 +/- 403 ng/ml; p less than 0.05 vs. controls) and in the group receiving daily CyA (5 mg/kg) +/- C (10 mg/kg) (2440 +/- 265 ng/ml; p less than 0.01 vs. controls); (ii) ALT and AST levels were increased in this latter group (ALT 223 +/- 133 UL, AST 114.67 +/- 39 UL; p less than 0.01 vs. controls); (iii) SC levels were normal; and (iv) steatosis of the liver was observed in these two groups. These findings suggest that C and R, but not F, may inhibit the hepatic cytochromes P-450 which are involved in the oxidative metabolism of the drugs. Furthermore, the high serum CyA levels seem to play a major role in the appearance of biochemical and histological damage to the liver. PMID- 2570687 TI - Effect of maprotiline combined with conventional neuroleptics against negative symptoms of chronic schizophrenia. AB - The therapeutic efficacy and target symptoms of maprotiline were tested by administering it in addition to conventional neuroleptic treatment for 10 weeks to a total of 32 chronic schizophrenic patients who showed no, or only partial, response to the neuroleptic medication. The final global improvement rating was 68.8% for all patients. Average therapeutic doses administered were 150 mg per day. Changes in psychotic symptoms were assessed by the Brief Psychiatric Rating Scale (BPRS), Psychiatric Evaluating scale (PES), and the Scale for the Assessment of Negative Symptoms (SANS). All mean improvement rates of these rating scales were observed at the 2nd week after the start of treatment, and maprotiline produced a marked amelioration in negative symptoms such as decreased spontaneity, blunted affect, emotional withdrawal, impaired work or recreation, etc. The incidence of side-effects was 37.5%. Constipation was the most frequently occurring side-effect. Neither side-effects nor laboratory test results were serious enough to discontinue the trial, except in the case of one chronic patient who showed acute exacerbation of symptoms due to maprotiline induced insomnia, elation and hallucination. These results suggest that maprotiline improves the negative symptoms of schizophrenia by a noradrenaline potentiating action not demonstrated by dopaminergic or serotonergic reward systems. PMID- 2570688 TI - Mechanisms of action of new antiepileptic drugs. AB - Our understanding of how new antiepileptic drugs work mirrors what we know about how currently marketed antiepileptic compounds exert their action--that information is scarce and elusive. The mechanism of action of antiepileptic drugs is nevertheless inextricably linked to epileptogenesis itself, and investigations of several promising new compounds are underway to establish the levels at which these drugs act. Compounds act on synapses and membranes as well as affecting receptors, neurotransmitters, and peptides. The most extensive data are available on drugs that inhibit the action of GABA or its receptors, including new benzodiazepine-like agents and barbituric-acid derivatives. The few drugs that act by inhibiting the effects of excitatory amino acids are reviewed. Finally, the maximal electroshock test is an empirical method to determine the antiepileptic properties of a drug; several agents under development have been effective in this screening technique. PMID- 2570689 TI - Designing molecules: specific peptides for specific receptors. AB - Peptides are the largest class of mediators of intercellular communication in the central nervous system. These molecules pose special problems in design for potential medical applications because of the high degree of flexibility, lack of high receptor selectivity, and ready biodegradation or clearance. The global and local use of conformational constraints has overcome these difficulties. Structure-biological activity relationships, molecular modeling, conformational analysis, conformational calculations, and molecular dynamics simulations can all be used to derive suitable lead structures and conformational models. Often, a single, constrained peptide analogue can be designed, which will have many of the desired biological and biophysical properties, and will serve as a template. Peptide analogues with high potency, exquisite receptor selectivity, and biological stability can be obtained. The approach is illustrated by the design of cyclic enkephalin-agonist analogues with exceptional delta-opioid-receptor selectivity, and of constrained somatostatin analogues that have become opioid peptides and possess potent opioid antagonist activities and exceptional selectivity for mu-opioid receptors. PMID- 2570690 TI - Mapping the gene for juvenile myoclonic epilepsy. AB - The practice of epileptology at a molecular level, where gene products are identified by gene mapping, will soon be possible for a growing number of epilepsies. Juvenile myoclonic epilepsy (JME) is the first of such epilepsies to be mapped to a chromosome, namely chromosome 6p21.3. Family studies of 68 JME probands from California revealed 50% of all families reported seizures in first- or second-degree relatives. Twelve percent of all family members other than the proband had epileptic seizures. Eighty percent of symptomatic siblings and 6% of asymptomatic siblings had diffuse 4- to 6-Hz multispike-wave complexes. Twelve percent of asymptomatic parents had diffuse, nonspecific slow waves mixed with spikes or sharp waves. JME is tightly linked to the Bf-HLA loci in chromosome 6. No matter what mode of inheritance is assumed, linkage to the clinical manifestations of JME and its associated EEG traits is indicated by lod scores over 3.0, as long as "EEG affected" but clinically asymptomatic family members are counted as affected during LIPED analysis. Studies are now being done to further localize the JME site. At the same time, further linkage studies should decide if JME is heterogeneous within itself and whether the same JME site in 6p21.3 underlies absence and grand mal epilepsies. PMID- 2570693 TI - Purification and characterization of an acetyl-CoA hydrolase from Saccharomyces cerevisiae. AB - Acetyl-CoA hydrolase, which hydrolyzes acetyl-CoA to acetate and CoASH, was isolated from Saccharomyces cerevisiae and demonstrated by protein sequence analysis to be NH2-terminally blocked. The enzyme was purified 1080-fold to apparent homogeneity by successive purification steps using DEAE-Sepharose, gel filtration and hydroxylapatite. The molecular mass of the native yeast acetyl-CoA hydrolase was estimated to be 64 +/- 5 kDa by gel-filtration chromatography. SDS/PAGE analysis revealed that the denatured molecular mass was 65 +/- 2 kDa and together with that for the native enzyme indicates that yeast acetyl-CoA hydrolase was monomeric. The enzyme had a pH optimum near 8.0 and its pI was approximately 5.8. Several acyl-CoA derivatives of varying chain length were tested as substrates for yeast acetyl-CoA hydrolase. Although acetyl-CoA hydrolase was relatively specific for acetyl-CoA, longer acyl-chain CoAs were also hydrolyzed and were capable of functioning as inhibitors during the hydrolysis of acetyl-CoA. Among a series of divalent cations, Zn2+ was demonstrated to be the most potent inhibitor. The enzyme was inactivated by chemical modification with diethyl pyrocarbonate, a histidine-modifying reagent. PMID- 2570691 TI - Liver enzyme changes in a guinea-pig model of facial eczema (sporidesmiotoxicosis). AB - Sporidesmin, a hepatotoxin from Pithomyces chartarum, is responsible for facial eczema in ruminants. In an attempt to clarify the biochemical processes supporting sporidesmin toxicity and response of the liver, haematology, plasma biochemistry and liver enzyme changes were monitored for 21 days in a model for facial eczema resulting from a single intraperitoneal injection of 2.8 mg/kg BW sporidesmin to guinea pigs. Most plasma disturbances were observed 8 days after administration and accounted for starvation, liver cytolysis, and cholestasis or liver enzyme induction. Alterations of hepatic enzyme activities were intense with a maximum increase on days 2 for alkaline phosphatases (ALP) and 8 for gamma glutamyltransferase (GGT), and a maximum decrease on day 21 for aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT). Comparison of liver and plasma enzyme changes indicates that GGT was the most reliable and significant plasma indicator of sporidesmin-associated liver alterations. Moreover, this study points out the validity of the one-dose intoxicated guinea pig model for research on sporidesmin biochemical toxicity and pathobiology of facial eczema. PMID- 2570692 TI - Hypercalcemia in the multiple endocrine neoplasia syndromes. AB - Multiple endocrine neoplasia includes disorders with hyperfunction of two or more endocrine tissues. In MEN type 1, hyperfunction of the parathyroid glands causing hypercalcemia is the most common clinical presentation. In vitro, suppression of parathyroid tissue by calcium is similar, but the set-point of hyperplastic tissue is shifted as compared with normal. The gene for MEN-1 has been localized to chromosome 11 and is linked to the basic fibroblast growth factor gene. Parathyroidectomy results in a high failure rate with recurrent hyperparathyroidism or autonomous graft function in autotransplanted tissue. Family screening is recommended once every 5 years in first-degree relatives. The approach to hyperparathyroidism in MEN-2 (2A) must be individualized during surgery for medullary thyroid carcinoma. Hyperparathyroidism in MEN-3 (2B) is often associated with normal serum calcium and may not require intervention. PMID- 2570694 TI - gamma-Glutamylcyclotransferase: inhibition by D-beta-aminoglutaryl-L-alanine and analysis of the solvent kinetic isotope effect. AB - Spin-echo NMR spectroscopy was used to record the cleavage of a gamma-glutamyl- amino-acid by (5-L-glutamyl)-L-amino-acid 5-glutamyltransferase (cyclizing) (gamma-glutamylcyclotransferase) in human erythrocyte hemolysates. The Michaelis Menten steady-state kinetic parameters were obtained by fitting the integrated Michaelis-Menten equation to the reaction time curves. The product, L-5 oxoproline, was shown to be an inhibitor of the reaction. The active site of the enzyme was probed by studies of the inhibition by D- and L-beta-aminoglutaryl-L alanine which are the beta-amino-acid isomers of D- and L-gamma-glutamyl-L alanine (the latter being a natural substrate of the enzyme); the D-isomer was the more potent inhibitor (Ki = 0.30 +/- 0.02 mmol/l water). When the alanyl alpha-carboxyl of the inhibitor was reduced to a hydroxyl (i.e. to give D-beta aminoglutaryl-L-alaninol) the potency of inhibition was reduced. The previously reported kinetic isotope effect of solvent 2H2O on the enzyme-catalyzed reaction has been further studied using a proton inventory. We propose that the solvent kinetic isotope effect is due to an intramolecular proton transfer between the glutamyl amino group and the peptide bond nitrogen. PMID- 2570696 TI - Preparative free flow electrophoresis for the isolation of two populations of proximal cells from the rabbit kidney. AB - High voltage free flow electrophoresis is a carrier-free method used for analytical and preparative cell separation, based on charge surface properties of cells. Two cell populations from the proximal tubule of the rabbit kidney were isolated by free flow electrophoresis from a suspension of pure proximal cells. This single-cell suspension was obtained through an original method by the combination of a Ca-binder action and gentle mechanical treatment associated with several shifting steps, on a pure suspension of isolated proximal tubules. Before the electrophoretic separation, the proximal cell origin was confirmed by enzymatic marker measurements, and the metabolic capacity was assessed by the cell respiratory activity. The isolated cells were very poor in distal tubule marker enzymes and were enriched in proximal tubule marker enzymes. Respiratory measurement showed a high cell metabolic capacity. After the electrophoretic separation, the origin of the cell populations was assessed by measuring specific marker enzymes. The cells in the slow-moving electrophoresis fractions had a high gamma-glutamyl transpeptidase activity and a low glucose-6-phosphatase activity. The fast moving cells showed a high glucose-6-phosphatase content and a poor gamma-glutamyl transpeptidase activity. Cells isolated by free flow electrophoresis were shown to possess long microvilli. This new methodology, allowed for the first time, the separation of a fast-moving cell population originating from the convoluted portion of the proximal tubule and a slow-moving cell population originating from the straight part of the proximal tubule of the rabbit kidney. PMID- 2570695 TI - Growth, morphology, function, and morphogenetic properties of rat renal glomerular epithelial cells in vitro: effects of retinyl acetate. AB - We report here that retinyl acetate (RA) regulates growth, morphology, function and cell organization in rat renal glomerular epithelial cells (SGE1). SGE1 cells are able to grow in a serum-free medium (DHFs medium) which is supplemented with insulin, transferrin, selenium, bovine serum albumin (BSA), linoleic acid and epidermal growth factor (EGF). When 0.1 and 1 micrograms/ml RA were added to the medium, the growth rates in the sparse culture were noticeably increased, compared to those in DHFs alone, whereas more than 10 micrograms/ml RA was cytotoxic to the cells. In the confluent culture, addition of 0.1, 1.0 and 10 micrograms/ml RA prolonged the cell survival. Since 10 micrograms/ml RA is not cytotoxic to the confluent culture, the cytotoxic action of RA seems to be dependent on cell density as well as RA dose. Ultrastructural observation revealed that RA treatment caused an increase of microvilli and alteration of cell shape, from flattened to columnar. Biochemical and immunological studies revealed that RA treatment increased the activity of r-glutamyl transpeptidase (GGT) and an amount of the membrane component with molecular mass (Mr) of 108,000 which is identical to one of nephritogenic antigens, Fx1A. By using fluorescence phalloidin stain, it was found that RA treatment increased content and organization of F-actin fibers. Furthermore, in collagen-embedding culture, RA induced 3-dimensional (3D) growth of SGE1 cells leading to the formation of organoids, cystic spheres with central lumen, in a serum-free condition; the addition of DHFs to collagen gel alone was ineffective for the 3D growth.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570697 TI - Technical considerations using a pulsed neodym-YAG laser for endoscopic shock wave lithotripsy. AB - First clinical experience with the use of a Q-switched pulsed Neodym-YAG Laser showed promising results. This paper focuses on two problems with respect to the optimal use of this laser: (1) is there any need for a special iron (Fe3+) enriched irrigant, and (2) what is the best frequency for laser lithotripsy? To answer these questions, we used an in vitro model, measuring the laser-induced breakdown (LIB) photographically utilizing sodium chloride as an irrigant enriched with different amounts of Fe3+ ions. The disintegrative efficacy of the laser was tested utilizing a standard stone model (plaster cube) and working at different frequencies (1, 10, 40 Hz). The addition of Fe3+ ions resulted in significant improvement of LIB. However, in the presence of a test stone no difference between sodium chloride and Fe3+-enriched irrigants was noted. The use of lower frequencies (1, 10 Hz) lead to a remarkable improvement in the disintegrative efficacy of the laser compared to the standard frequencies (40, 50 Hz). For clinical use, addition of Fe3+ ions seems only necessary if optical breakdown (LIB) is insufficient despite the increase in generator voltage. In such a situation, we recommend the addition of an 1-ml Fe3+ solution to 10 liters of sodium chloride irrigant (= 0.5 mg Fe3+/dl). Moreover, the standard frequency for laser-induced intracorporeal lithotripsy should be 1-10 Hz. PMID- 2570699 TI - Renal and hormonal effects of alpha 1-adrenoceptor blockade by bunazosin in essential hypertension. AB - The renal and hormonal effects of the alpha 1-adrenoceptor blocker bunazosin were examined in 6 patients with essential hypertension. Oral bunazosin for 4 to 12 weeks significantly decreased mean blood pressure by 10%, increased effective renal blood flow and creatinine clearance by 34% and 37%, respectively, the plasma norepinephrine concentration was elevated by 60%, and the plasma atrial natriuretic peptide level was lowered by 22%. The plasma renin activity and aldosterone concentration were unchanged. Thus, a moderate reduction in blood pressure was produced by bunazosin treatment while maintaining renal perfusion. PMID- 2570698 TI - Clinical significance of the sparteine/debrisoquine oxidation polymorphism. AB - The sparteine/debrisoquine oxidation polymorphism results from differences in the activity of one isozyme of cytochrome P450, the P450db1 (P450 IID1). The oxidation of more than 20 clinically useful drugs has now been shown to be under similar genetic control to that of sparteine/debrisoquine. The clinical significance of this polymorphism may be defined by the value of phenotyping patients before treatment. The clinical significance of such polymorphic elimination of a particular drug can be analyzed in three steps: first, does the kinetics of active principle of a drug depend significantly on P450db1?; second, is the resulting pharmacokinetic variability of any clinical importance?; and third, can the variation in response be assessed by direct clinical or paraclinical measurements? It is concluded from such an analysis that, in general, the sparteine/debrisoquine oxidation polymorphism is of significance in patient management only for those drugs for which plasma concentration measurements are considered useful and for which the elimination of the drug and/or its active metabolite is mainly determined by P450db1. At present, this applies to tricyclic antidepressants and to certain neuroleptics (e.g. perphenazine and thioridazine) and antiarrhythmics (e.g. propafenone and flecainide). Phenotyping should be introduced in to clinical routine under strictly controlled conditions to afford a better understanding of its potentials and limitations. The increasing knowledge of specific substrates and inhibitors of P450db1 allows precise predictions of drug-drug interactions. At present, the strong inhibitory effect of neuroleptics on the metabolism of tricyclic antidepressants represents the best clinically documented and most relevant example of such an interaction. PMID- 2570700 TI - Evaluation of the beta-adrenoceptor blocking activity of the Class Ic antiarrhythmic drug diprafenone in man. AB - The beta-adrenoceptor blocking activity of the new Class Ic-antiarrhythmic drug diprafenone has been determined in man in comparison with that of propranolol, using the standardized isoproterenol test. Following placebo, the dose of isoproterenol which increased the heart rate by 25 beats.min-1 (CD25) was 0.84 (0.1-2.72) micrograms (median and range). Two hours following single oral doses of 200 mg diprafenone (3.61 micrograms; 1.53-44.15) or 40 mg propranolol (16.06 micrograms; 9.15-27.04) significantly higher CD25-values were found. The affinity constants for the beta-adrenergic receptors calculated on the basis of free drug in the plasma were similar for propranolol and diprafenone (2.79 vs. 2.60 nM-1, respectively). Thus, diprafenone showed a definite degree of beta-blocking activity in a clinically relevant range of plasma concentrations. PMID- 2570701 TI - Suppression of histamine-induced skin reactions by loratadine and cetirizine diHCl. AB - The skin reactivity to histamine was measured in 10 healthy and 10 atopic subjects after the ingestion of single doses of loratadine 10 mg, cetirizine diHCl 10 mg and placebo, in a double-blind cross-over randomized study. The anti H1 effect of cetirizine diHCl proved to be significantly more rapid, more pronounced and longer lasting than that of loratadine. PMID- 2570703 TI - Murine enterocytes can present soluble antigen to specific class II-restricted CD4+ T cells. AB - Highly purified mature epithelial cells (EC) from murine small intestinal villi (excluding Peyer's patch epithelium) were examined for their capacity to present foreign antigen to T cells. Cell suspensions composed of 99% Ia+T200- EC were able to present keyhole limpet hemocyanin to an antigen-specific class II restricted L3T4+ T cell hybridoma, and stimulate interleukin 2 production. Antigen presentation by EC was inhibited by major histocompatibility complex class II-specific monoclonal antibodies. It is concluded that EC express functional class II molecules and may activate adjacent CD4+ T cells to induce lymphokine synthesis. PMID- 2570702 TI - Complex Ca2+ flux inhibition as primary mechanism of staurosporine-induced impairment of T cell activation. AB - The inhibitory effect of the highly effective drug staurosporine on the early activation signal Ca2+ flux was investigated via multiparameter flow cytometry in human peripheral blood T lymphocytes. Staurosporine has been reported to be a specific inhibitor of protein kinase C. However, we show that it inhibits the Ca2+ influx in anti-CD3 and phytohemagglutinin-stimulated human CD4+ and CD8+ lymphocytes at concentrations between 1.0 and 10.0 ng/ml. Staurosporine decreases the number of Ca2+-positive CD4+ and CD8+ lymphocytes as well as the Ca2+ influx per cell; the drug also delays the time of the maximum response to polyclonal stimulation. In addition, we demonstrate that staurosporine affects the primary Ca2+ response via inhibition of the release of the membrane-bound Ca2+ from the endoplasmic reticulum in CD4+ and CD8+ lymphocytes. Binding studies of the anti CD3 antibody to T lymphocytes indicate normal binding capacities in the presence of staurosporine. With respect to the classical scheme of T cell activation via phospholipase C, our data suggest that staurosporine may inhibit T cell activation primarily by its effect on the early Ca2+ flux signal. PMID- 2570704 TI - Effect of MK-801 on the decrease in tryptophan hydroxylase induced by methamphetamine and its methylenedioxy analog. AB - The role of N-methyl-D-aspartate (NMDA) receptors in the decrease in neostriatal tryptophan hydroxylase (TPH) activity induced by repeated high doses of methamphetamine or 3,4-methylenedioxymethamphetamine (MDMA) was evaluated. Rats received 4 injections of methamphetamine (15 mg/kg) or MDMA (10 mg/kg) at 6 h intervals, and were killed 18-20 h after the last administration. These treatments with methamphetamine or MDMA reduced neostriatal TPH activity to 26 and 34% of control, respectively. Coadministration of MK-801 (2.5 mg/kg) significantly attenuated the methamphetamine-induced decrease in TPH activity (66% of control), but did not alter the effect of MDMA. This study suggests that excitatory amino acids may participate in the methamphetamine-induced decline in central TPH activity, and that the mechanism by which MDMA and methamphetamine decreases TPH activity may differ. PMID- 2570706 TI - [3H]RX821002: a new tool for the identification of alpha 2A-adrenoceptors. AB - The human adenocarcinoma cell-line HT29 was used as a model to investigate the binding properties of a new antagonist radioligand of the imidazoline series, [3H]RX821002. All aspects of [3H]RX821002 binding conclusively prove that this radioligand is a valuable tool for labelling alpha 2A-adrenoceptors. [3H]RX821002 binding was very rapid and reversible. Computer-assisted analysis of kinetic data revealed association and dissociation time courses consistent with a simple bimolecular reaction. Saturation isotherms indicated that [3H]RX821002 labeled with high affinity a single population of non-interacting sites displaying a KD of 1.7 +/- 0.1 nM. Adrenoceptor agonists and antagonists inhibited [3H]RX821002 and [3H]yohimbine binding with a strictly similar rank order of potency which is characteristic of alpha 2A-adrenoceptors. The binding parameters of [3H]RX821002 were compared with those of other commercially available [3H]antagonists, [3H]yohimbine and [3H]idazoxan. Analysis of the saturation isotherms for the three radioligands showed that (1) [3H]RX821002 was the radioligand exhibiting the lower percentage of non-specific binding and the better affinity, (2) the Bmax of [3H]RX821002 was significantly higher than that of [3H]yohimbine. The difference in Bmax was not due to better labelling of one of the two affinity states of the receptor but was greatly reduced in glycylglycine buffer, suggesting that, in Tris-Mg2+ buffer, [3H]yohimbine does not label the entire alpha 2-adrenoceptor population. PMID- 2570707 TI - Influence of dopamine receptor agonists and antagonists on calmodulin translocation in different brain regions. AB - Acute and chronic experiments were performed to study the effects of intraperitoneally administered dopamine receptor agonists and antagonists on the translocation of cytosolic and membrane-bound calmodulin in the striatum and hippocampus of the rat brain. Single doses of apomorphine and a low-dose amphetamine (1.25 mg/kg) resulted in a translocation of calmodulin, as measured by a decrease in membrane-bound and increase in cytosolic calmodulin in the striatum, whereas bromocryptine was ineffective. Amphetamine exerted a similar effect in the hippocampus and striatum. However, a high-dose amphetamine (5 mg/kg) had an opposite effect on translocation, in that there was an increase in membrane-bound calmodulin. Chronically applied amphetamine (5 mg/kg) and haloperidol (1 mg/kg), i.e. under conditions of dopamine receptor supersensitivity, tended to decrease cytosolic calmodulin in the striatum and hippocampus, and to increase membrane-bound calmodulin. The findings and, especially, the chronic effects of haloperidol and the high dose of amphetamine are interpreted in the light of the current concept which suggests that transmitters and intraneuronal signalling systems converge, thereby influencing the more complex processes of neuronal plasticity, rather than receptor sensitivity only. PMID- 2570705 TI - In vivo experiments for the evaluation of alpha 1-adrenoceptor antagonistic effects of SGB-1534 on canine urethra. AB - The alpha 1-adrenoceptor blocking effects of SGB-1534 on the urethral smooth muscle were compared in in vivo lower urinary tract preparations of anesthetized dogs, with the effects of other alpha 1-adrenoceptor antagonists, prazosin and bunazosin. Hypogastric nerve stimulation and selective administration of phenylephrine to the urethra and bladder through the cannulated right external iliac artery (i.a.) elicited reproducible frequency- and dose-dependent increases in intra-urethral pressure. Intra-bladder pressure was increased by the nerve stimulation but not by i.a. phenylephrine. SGB-1534, prazosin or bunazosin (0.1 10 micrograms/kg i.v.) dose dependently suppressed the urethral contraction evoked by the nerve stimulation and i.a. phenylephrine but did not influence the bladder contraction elicited by nerve stimulation. The alpha 1-adrenoceptor blocking potency of SGB-1534 was approximately 2.3 and 8.1 times greater than that of prazosin and bunazosin, respectively. The results indicate that alpha 1 adrenoceptors may mediate mainly the urethral contraction induced by hypogastric nerve stimulation and i.a. phenylephrine, and that SGB-1534 was more potent alpha 1-adrenoceptor blocking activity than prazosin and bunazosin in the canine urethra. PMID- 2570709 TI - Molecular characterization of size and copy number of the human insulin gene in transfected rat islet tumor cell clones. AB - Two stable transfectants, NHI-6F and NHI-5B, obtained by cotransfection of the human insulin gene (a 15 kb EcoRI genomic fragment) with pSV2-neo into MSL-cells (pluripotent rat islet tumor cells), were characterized with regard to copy number and size of the inserted human sequences by Southern blot analyses using appropriate restriction enzymes. Digestion of total genomic DNA from the NHI-6F clone with EcoRI, SacI and BglII gave in all cases single human insulin gene fragments of 25 kb, 28 kb and 11 kb, respectively. From these data we were able to conclude that NHI-6F carries a single copy of the human insulin gene and that this copy has at least retained the 5' BglI and the 3' BglII flanking sites of human origin. Digestion of NHI-5B DNA gave a different banding pattern compared to NHI-6F (EcoRI: 20 kb; SacI: 7.8 kb; BglII: 11 kb). These data indicate the presence of a single human insulin gene copy in clone NHI-5B with the preservation of the 5' SacI and 3' BglII human flanking sites. In conclusion, the two transfected rat islet tumor clones, NHI-6F and -5B, carry a single copy of the human insulin gene. Each copy is presumably functionally active since known promoter and enhancer elements reside within 400 bp 5' to the gene. NHI-6F and 5B carry 2,500 bp and 4,500 bp 5' to the gene, respectively. PMID- 2570708 TI - Pancreatic islets obtained from pancreases with short-term exocrine atrophy disturbed morphology but normal in vitro response to various stimuli. AB - The morphology and function islets isolated from rat pancreases with short-term (16 days) exocrine atrophy was investigated. It was found that this type of atrophy induced changes in the shape and morphological structure of pancreatic islets. However, the function of isolate islets did not change, as investigated by the basal or glucose-stimulated secretion of insulin, glucagon and somatostatin, and by the CCK-stimulated secretion of insulin. We conclude that the rat pancreas brought to atrophy by a short-term ligation of the pancreatic duct can be considered as a rich source of functionally viable islets. PMID- 2570710 TI - Glucoregulatory function of thyroid hormones: evidence for an insulin-independent effect. AB - The effect of hyperglycemia on glucose kinetics was investigated in normal and T4 treated miniature pigs. 1. T4-treatment increased basal plasma glucose (G) (+ 20%) and glucose turnover (+36%). 2. Glucose infusion (2 mg/kg x min) in controls increased insulin and glucose utilization (Rd), but decreased glucagon and hepatic glucose production (Ra). After T4-treatment glucose increased insulin, decreased glucagon and Ra but only a slight effect on G and Rd were observed. 3. Infusing glucose + somatostatin resulted in hyperglycemia in both groups due to an initial fall in Ra and Rd followed by an increase in Rd, where total Ra (endogenous + exogenous exceeded Rd. Glucose intolerance was more pronounced in controls, due to a T4-induced increase in Rd. During this non-steady state the increment in Rd per increment in G was calculated and showed 1.5 in controls and 2.5 after T4-treatment. These data give evidence that thyroid hormones increase glucose utilization during hypoinsulinemia. PMID- 2570711 TI - Antilymphocytic antibodies and marrow transplantation. IX. T-cell depletion in marrow donors with C1q high and low affinity antibodies for suppression of GVHD in fully mismatched mice. AB - Rat monoclonal antibodies (mAbs) of the same specificity (anti-Thy-1) but different immunoglobulin (Ig) subclass were investigated for their effect on suppression of graft-versus-host disease (GVHD) by depleting the marrow donors of T cells in vivo. Transplantation to homozygous, fully mismatched mice of spleen and bone marrow cells from unthymectomized mice injected with the mAbs revealed that two rat anti-Thy-1 mAbs with high affinity for C1q (IgG2b) suppressed and prevented acute and chronic mortality of GVHD. In contrast, rat mAbs with low affinity for C1q (IgM, IgG2c) barely delayed acute mortality. This correlated with findings on the degree of splenic T-cell depletion in donor mice with the IgG2b mAb, able to deplete 97%, and the IgG2c and IgM mAbs, only 83% and 75% of T cells, respectively. An effect akin to the one achieved with IgG2b was seen, however, when donor mice were thymectomized and then treated with three injections of IgG2c isotype. The rat IgM mAb was not immunosuppressive even under such conditions. Immunocytochemical and immunohistochemical examination of the donor lymph nodes after single injection of either mAb showed that only 84% of T cells were eliminated, and in contrast to the spleen, none of the tested antibodies could deplete T cells further. The thymus did not appear depleted at all, although the cortical thymocytes were coated with either of the injected mAbs. PMID- 2570712 TI - Long-term culture of peripheral blood stem cells: the effect of the addition of an irradiated stromal layer. AB - We have studied peripheral blood stem cells (PBSC) collected by cytapheresis following intensive chemotherapy, from 13 patients with acute leukemia, in long term culture (LTC). Peripheral blood was cultured with (n = 10) and without (n = 21) the addition of a preformed, irradiated stromal layer. In this latter LTC our results confirm that peripheral blood is capable of producing CFU-GM and nucleated cells in the absence of the formation of an adherent stromal layer. However, peripheral blood cultured in the presence of an irradiated stromal layer is capable of a significantly higher proliferative response (total production of CFU GM per flask - mean = 57529) than in the absence of an irradiated stromal layer (total production of CFU GM per flask - mean = 26739, p less than 0.03). Our results suggest that PBSC contain a primitive nonplastic adherent cell that requires the presence of a stromal layer for its expression. These findings provide further support for the use of peripheral blood stem cells for autologous transplantation. PMID- 2570716 TI - High doses of sympathomimetics in severe bronchial asthma. PMID- 2570715 TI - Possible existence of a presynaptic positive feedback mechanism enhancing dopamine transmission in the anterior cingulate cortex of the rat. AB - A series of microiontophoretic and VTA stimulation experiments, conducted in intact, GBR-12909-treated, alpha-methylparatyrosine-depleted or 6-hydroxydopamine denervated rats, provide suggestive evidence for the existence of a presynaptic, positive feedback mechanism triggered by dopamine reuptake and favoring the release of this transmitter in the anterior cingulate cortex. PMID- 2570717 TI - The relation between snoring and smoking, body mass index, age, alcohol consumption and respiratory symptoms. AB - Potential risk factors for snoring were studied in a population of 457 middle aged men. Eversnoring was reported by 60% of the men and snoring with an age of onset before or equal to 20 years by 13%. Eversnoring was significantly related to older age, higher body mass index and smoking habits. Alcohol consumption, estimated by questionnaire and gamma-glutamyl transpeptidase was unrelated to a history of snoring. Logistic regression showed that snoring was independently associated with age, body mass index and smoking habits. An exposure-effect relationship clearly appeared between tobacco consumption and snoring. After adjustment for smoking habits, none of the upper or lower respiratory symptoms was significantly related to snoring. PMID- 2570714 TI - Genetic control of hippocampal cholinergic and dynorphinergic mechanisms regulating novelty-induced exploratory behavior in house mice. AB - Neurobehavioral genetics endeavors to trace the pathways from genetic and environmental determinants to neuroanatomical and neurophysiological systems and, thence, to behavior. Exploiting genetic variation as a tool, the behavioral sequelae of manipulating these neuronal systems by drugs and antisera are analyzed. Apart from research in rats, this paper deals mainly with the genetically-influenced regulation in mice of exploratory behaviors that are adaptive in novel surroundings and are hippocampally-mediated. Special attention is paid to neuropeptidergic, GABAergic, and cholinergic synaptic functions in the mouse hippocampus. The behaviorally different inbred mouse strains C57BL/6 and DBA/2 show opposite reactions (reductions and increases, respectively, in exploration rates) to peripheral and intrahippocampal injections with agents that interfere with peptidergic, cholinergic, and GABAergic neurotransmission. These findings can be explained by an interdependent over-release of opioids, arrested GABA release, and excess acetylcholine in the hippocampal neuronal network of DBA/2 mice, as compared to C57BL/6 mice where these systems are functionally well balanced. Very similar results have been obtained with the lines SRH and SRL, derived from C57BL/6 and DBA/2, and genetically selected for rearing behavior. Most probably, the opioids act to disinhibit exploratory responses. An additional genetic approach is mentioned, in which four inbred mouse strains and one derived heterogeneous stock are used for estimating genetic correlations between structural properties of the hippocampal mossy fibers and levels of hippocampal dynorphin B, on the one hand, and frequencies of exploratory responses to environmental novelty, on the other. PMID- 2570718 TI - AVP receptors of mouse Leydig cells are regulated by LH and E2 and influenced by experimental cryptorchidism. AB - Exposure of pubertal mouse Leydig cells for 24 h to increasing concentrations (1 100 ng/ml) of LH elicited a dose-dependent decrease in AVP receptor content. Maximal reduction (50%) was obtained at a dose of 10 ng/ml LH. A similar treatment applied to adult Leydig cells did not influence AVP receptor density. Treatment of adult Leydig cells for 24 h by E2 (5-500 ng/ml) resulted in a dose dependent increase in AVP receptor content. About 50% increase was achieved with 500 ng/ml E2. AVP receptor content in pubertal Leydig cells was not modified irrespective of the concentration of E2 tested. These changes in AVP receptor number were well correlated with the response of Leydig cells to AVP (10(-6) M) in terms of testosterone production. 2 weeks bilateral cryptorchidism resulted in reduction of testicular weight, circulating testosterone levels associated with a marked rise in Leydig cell AVP receptor density with no change of affinity. Testosterone production by Leydig cells from cryptorchid testes in response to AVP (10(-6) M) or hCG (100 ng/ml) stimulation was reduced compared to that of control Leydig cells. This study provides new arguments supporting the concept that AVP could be involved in local regulation of testicular steroidogenesis. PMID- 2570713 TI - Selected mouse lines, alcohol and behavior. AB - The technique of selective breeding has been employed to develop a number of mouse lines differing in genetic sensitivity to specific effects of ethanol. Genetic animal models for sensitivity to the hypnotic, thermoregulatory, excitatory, and dependence-producing effects of alcohol have been developed. These genetic animal models have been utilized in numerous studies to assess the bases for those genetic differences, and to determine the specific neurochemical and neurophysiological bases for ethanol's actions. Work with these lines has challenged some long-held beliefs about ethanol's mechanisms of action. For example, lines genetically sensitive to one effect of ethanol are not necessarily sensitive to others, which demonstrates that no single set of genes modulates all ethanol effects. LS mice, selected for sensitivity to ethanol anesthesia, are not similarly sensitive to all anesthetic drugs, which demonstrates that all such drugs cannot have a common mechanism of action. On the other hand, WSP mice, genetically susceptible to the development of severe ethanol withdrawal, show a similar predisposition to diazepam and phenobarbital withdrawal, which suggests that there may be a common set of genes underlying drug dependencies. Studies with these models have also revealed important new directions for future mechanism-oriented research. Several studies implicate brain gamma-aminobutyric acid and dopamine systems as potentially important mediators of susceptibility to alcohol intoxication. The stability of the genetic animal models across laboratories and generations will continue to increase their power as analytic tools. PMID- 2570719 TI - Reduction in the size of prolactin-producing pituitary tumor after Cabergoline administration. AB - Different weekly doses (400 to 3,000 micrograms) of the new, long-acting dopamine agonist Cabergoline (Farmitalia Carlo Erba, Milan, Italy) were given to 11 hyperprolactinemic women with pituitary tumor. Pituitary computerized tomography (CT) scans were performed before the start of treatment and after 3 (n = 5), 6 (n = 3), and 9 (n = 3) months of Cabergoline administration. Plasma prolactin (PRL) was determined in blood samples collected before and at weekly intervals during Cabergoline administration. Cabergoline induced marked inhibition of PRL secretion in conjunction with a CT demonstration of reduction in the pituitary tumor size in all patients. The potent, long-lasting PRL inhibitory effect of Cabergoline and the absence of side effects typical of dopaminergic compounds suggest that the use of this drug is advantageous over others in the medical treatment of hyperprolactinemia. PMID- 2570720 TI - Oxytocin mRNA: increase of polyadenylate tail size during pregnancy and lactation. AB - Earlier studies indicate that rat hypothalamic oxytocin (OT) mRNA accumulation rises gradually during pregnancy and remains elevated throughout the lactation period. Here we show that, in addition, hypothalamic OT mRNA undergoes a structural change during this period. On gel electrophoresis, the size of rat OT mRNA increased from 750 bases (control) to a maximum of 820 bases (7th day of lactation). Removal of the poly(A) tail by the RNase H methods revealed that this size increase can be fully accounted for by a prolongation of the polyadenylate tail. There is considerable evidence for a role of the poly(A) tail segment in augmenting mRNA stability and, possibly, translational efficiency. It is thus conceivable that the demonstrated regulation of OT mRNA poly(A) tail length represents an additional level of OT gene control during pregnancy and lactation. PMID- 2570722 TI - Multiple functions of a Drosophila homeotic gene, zeste-white 3, during segmentation and neurogenesis. AB - Lack of both maternal and zygotic gene activity at the zeste-white 3 (zw3) locus causes severe developmental transformations. Embryos derived from germ cells that lack zw3+ gene activity die during embryogenesis and have a phenotype that is similar to that of embryos mutant for the segment polarity gene naked (nkd). In both nkd and germ line clone-derived zw3 embryos the pattern elements derived from the anterior-most part of each segment, the denticle belts, are deleted. Similar abnormal patterns of the zygotically expressed genes engrailed and Ultrabithorax are detected in both mutants, suggesting that the two genes are involved in the same developmental process. Additionally, the induction of clones of zw3 mutant cells in imaginal discs causes homeotic transformations of noninnervated hair cells into innervated sensory bristles. The multiple roles of zw3 during development and its possible interactions with the zygotic gene nkd are discussed. PMID- 2570721 TI - Ultrastructural distribution of growth hormone (GH) mRNA and GH intron I sequences in rat pituitary gland: effects of GH releasing factor and somatostatin. AB - We have measured the distribution of growth hormone (GH) mRNA or intron I sequences by in situ hybridization on ultrathin frozen sections of pituitaries removed from rats injected with saline, GH-releasing factor (GRF) or somatostatin. A 4-fold increase in labeling of the anterior lobe was observed after GRF, no changes with somatostatin. After ultrastructural in situ hybridization, labeling with the GH cDNA probe was specific to somatotrophs. Two populations of cells containing few or many secretory granules were labeled mainly in the cytoplasm or in both cytoplasm and nucleus. Some cells showed labeling at the perinuclear membrane. Hybridization with the GH intron I probe showed the same cell specificity with silver grains mainly located in the nucleus. After GRF, sequences hybridizing to growth hormone cDNA were increased mainly in the nucleus of somatotrophs when compared to mock-injected rats, indirectly suggesting an increase in the transcriptional activity of the growth hormone gene. After somatostatin, the density of labeling in the nucleus was increased suggesting that somatostatin may prevent the export of growth hormone mRNA molecules from the nucleus to the cytoplasm. PMID- 2570723 TI - A genetic and developmental analysis of mutations in labial, a gene necessary for proper head formation in Drosophila melanogaster. AB - We have undertaken a developmental genetic analysis of labial (lab), the most proximal gene in the Antennapedia complex (ANT-C) of Drosophila melanogaster. The terminal phenotype of mutant embryos was examined in cuticle preparations, in thin sections, and by scanning electron microscopy. These preparations revealed a failure of head involution and the loss or disruption of several head structures, including the salivary glands and the H-piece and ventral arm of the cephalopharyngeal apparatus. Although these structures are presumed to derive from the gnathocephalic segments, we argue that the observed defects are likely to be a secondary consequence of a failure of head involution. A function for lab in the development of the adult head was inferred from the phenotype of animals bearing hypomorphic alleles and from clones of lab- tissue generated by mitotic recombination. Two aspects of the mutant phenotype were manifested. Ventrally, a deletion and/or disruption of tissue occurred in the maxillary palp and vibrissae regions. Dorsally, the posterior head appeared to be transformed to a thoracic like identity. Mutations in lab, like those in the Deformed and proboscipedia loci of the ANT-C, reveal a homoeotic phenotype only in the adult stage of the life cycle. PMID- 2570724 TI - Differential expression of human HOX-2 genes along the anterior-posterior axis in embryonic central nervous system. AB - We have investigated the structure of the human HOX-2 locus, which encompasses a 90-kb region on chromosome 17q21. Five new human HOX-2 homeoboxes, termed HOX 2.5, 2.4, 2.6, 2.7 and 2.8, have been identified, and their nucleotide sequences are reported. They have the same 5'-3' transcriptional orientation and are clustered with three previously described HOX-2 homeoboxes (5'-2.5-2.4-2.3-2.2 2.1-2.6-2.7-2.8-3'). We have also investigated the region-specific expression of HOX-2 genes in human embryonic-fetal life by Northern-blot analysis. All genes are expressed in whole embryos and fetuses at 5-9 weeks from conception. Their major site of expression lies within the central nervous system (CNS), although they are transcribed at a lower level in body structures other than the CNS. Their relatively abundant expression in CNS has been analyzed along the anterior posterior axis by dissecting the brain, the medulla oblongata and the spinal cord proper. HOX-2.5, 2.4 and 2.3 transcripts are markedly more abundant in spinal cord than in medulla, whereas 2.2, 2.1, 2.6 and 2.7 mRNAs are progressively more abundant in the medulla. Additionally, expression in brain was detected, although at lower level, for HOX-2.1, 2.6, 2.7, 2.8. Thus, the relative position of HOX-2 homeobox genes along the chromosome in the 5'-3' direction appears to correlate with the relative position of their expression domains along the CNS longitudinal axis in the caudal-cephalic direction. PMID- 2570726 TI - This is FASEB. The Research Conference Office, Education, and the Minority Program. PMID- 2570725 TI - Role of reversible phosphorylation of acetyl-CoA carboxylase in long-chain fatty acid synthesis. AB - Acetyl-CoA carboxylase, the rate-limiting enzyme in the biogenesis of long-chain fatty acids, is regulated by phosphorylation and dephosphorylation. The major phosphorylation sites that affect carboxylase activity and the specific protein kinases responsible for phosphorylation of different sites have been identified. A form of acetyl-CoA carboxylase that is independent of citrate for activity occurs in vivo. This active form of carboxylase becomes citrate-dependent upon phosphorylation under conditions of reduced lipogenesis. Therefore, phosphorylation-dephosphorylation of acetyl-CoA carboxylase is the enzyme's primary short-term regulatory mechanism; this control mechanism together with cellular metabolites such as CoA, citrate, and palmitoyl-CoA serves to fine-tune the synthesis of long-chain fatty acids under different physiological conditions. PMID- 2570727 TI - Measurement of immunoreactive gamma-glutamyl transpeptidase in human sera and liver tissues of patients with various liver diseases. AB - Immunoreactive gamma-glutamyl transpeptidase in human serum and liver tissue was measured by a solid phase enzyme-linked immunosorbent assay. The immunoreactive gamma-glutamyl transpeptidase was significantly elevated in the sera of patient with hepatocellular carcinoma. On the other hand, in sera of patients with non neoplastic diseases, including chronic hepatitis, acute hepatitis, fatty liver and hemangioma, the immunoreactive gamma-glutamyl transpeptidase was not elevated. In hepatocellular carcinoma and metastatic liver tumor tissues, the immunoreactive gamma-glutamyl transpeptidase content was also elevated, showing good correlation with the enzyme protein content in sera. However, no correlation was found between the activity of gamma-glutamyl transpeptidase determined by an enzymatic assay and the content determined by an enzyme-linked immunosorbent assay. On immunohistochemical examination, the immunoreactive enzyme protein without enzymatic activity was detected only in the cytoplasm of cancer cells. This suggested that there is an increased level of the immunologically active but enzymatically inactive form of gamma-glutamyl transpeptidase in hepatoma tissues. PMID- 2570728 TI - Effect of acute suppression of acid secretion by omeprazole on postprandial gastrin release in conscious dogs. AB - The effect of acute suppression of acid secretion induced by administration of a single dose of omeprazole (2 mg/kg body wt) on postprandial gastrin release was studied in 10 conscious dogs. In omeprazole-treated dogs, a sustained gastrin release was observed during a 10-h period after feeding, although greater than 95% of the meal had left the stomach after 4 h. This sustained gastrin release could be inhibited by acidification of the gastric lumen, by somatostatin, and by atropine. Insulin and bombesin induced considerable gastrin release in omeprazole treated dogs, but plasma gastrin concentrations returned almost to basal values after 3 h. Omeprazole administered alone had no significant effect on basal gastrin levels. These data indicate that, in dogs, when acid secretion is suppressed by omeprazole a meal induces a sustained gastrin release lasting for up to 10 h. This gastrin release is probably related to the fact that food has been in contact with the gastric lumen, as neither vagal nor bombesin stimulation induced such a sustained activity of the G cells. PMID- 2570729 TI - Enprostil reduces the increase of gastric corpus mucosal mass induced by the hydrogen-potassium-stimulated adenosine triphosphatase inhibitor BY 831-78 in the rat. AB - We studied whether enprostil, a synthetic prostaglandin E2 derivative, might inhibit gastrin release and the trophic effects on gastric oxyntic mucosa induced by prolonged treatment with an inhibitor of hydrogen-potassium-stimulated adenosine triphosphatase, the substituted benzimidazole BY 831-78. Rats were treated intragastrically with enprostil (1 or 15 micrograms/kg b.i.d.), BY 831-78 (15 mumol/kg once daily), the combination of enprostil and BY 831-78, ranitidine (300 mumol/kg b.i.d.), and placebo. Plasma gastrin and somatostatin levels and gastric acid secretion were measured during a 1-day treatment in animals fitted with chronic gastric fistulas and repeatedly during 9 wk of treatment in intact rats. Despite inhibiting acid secretion, enprostil did not increase plasma gastrin. When combined with BY 831-78, enprostil transiently reduced the BY 831 78-induced increase of integrated plasma gastrin (1375 +/- 206 vs. 2137 +/- 256 pmol/L.12 h, p less than 0.05) in fasted rats with fistulas, but failed to prevent the marked hypergastrinemia following 9 wk of treatment with BY 831-78 (717 +/- 80 vs. 731 +/- 56 pmol/L) in intact rats. However, enprostil reduced the BY 831-78-induced increase of oxyntic mucosal volume (458 +/- 31 vs. 567 +/- 33 mm3, p less than 0.01), whereas BY 831-78 prevented the enprostil-induced increase of antral mucosal volume (42 +/- 3 vs. 56 +/- 3 mm3, p less than 0.01). These results demonstrate that some of the trophic effects induced by a hydrogen potassium-stimulated adenosine triphosphatase inhibitor are not exclusively governed by gastrin. PMID- 2570731 TI - Relationship between gastric acid and elevated plasma somatostatinlike immunoreactivity after a mixed meal. AB - The aim of this study was to examine whether hydrochloric acid plays a role mediating the post-prandial increase in plasma somatostatinlike immunoreactivity in normal subjects. Intravenous infusion of cimetidine was found to reduce by 45% the postprandial increment in plasma somatostatin-like immunoreactivity. This effect was reversed by concomitant intragastric administration of 0.1 N hydrochloric acid, which in previous studies in fasted subjects had not affected plasma somatostatinlike immunoreactivity. The effects of cimetidine on postprandial plasma gastrin were the inverse of those observed on postprandial somatostatin. There was a greatly enhanced increment in postprandial plasma gastrin during cimetidine infusion, which was reduced significantly toward control levels by concomitant intragastric infusion of hydrochloric acid. To exclude direct inhibition by cimetidine of nutrient-stimulated plasma somatostatinlike immunoreactivity we studied the effect of cimetidine on plasma somatostatinlike immunoreactivity stimulated by an intraduodenal infusion of fat. Cimetidine did not alter the incremental response of somatostatinlike immunoreactivity to intraduodenal fat infusion. These data show that cimetidine does not invariably reduce nutrient-stimulated plasma somatostatinlike immunoreactivity and are consistent with the hypothesis that the action of cimetidine in reducing the plasma somatostatin response to ingestion of a meal is a consequence of reduction of postprandial acid secretion. These data suggest that the postprandial elevation in plasma somatostatin observed in humans is mediated in part through postprandial secretion of gastric acid, which in turn acts to elevate plasma somatostatin. PMID- 2570730 TI - Suppression of nocturnal acid secretion with famotidine accelerates gastric ulcer healing. AB - Although nocturnal acid secretion has been emphasized in the pathophysiology and treatment of duodenal ulcer, its importance in gastric ulcer disease has been questioned. To explore this area, this multicenter U.S. trial compared the effect of a once-daily nighttime dose of H2-receptor antagonist with placebo on the healing of gastric ulcer and relief of associated symptoms. One hundred fifty seven patients with endoscopically verified benign gastric ulcers were randomized in a double-blind fashion to either famotidine (40 mg at bedtime) or placebo. Antacid tablets were allowed as needed. The healing rates for famotidine were 45%, 66%, and 78% at weeks 4, 6, and 8, respectively. In comparison, placebo healing rates were 39%, 44%, and 64%. These differences were statistically significant in favor of famotidine at weeks 6 (p less than or equal to 0.01) and 8 (p less than or equal to 0.05), as well as in a life-table analysis (p less than or equal to 0.05). Nocturnal famotidine was also significantly better than placebo with respect to time to complete relief of pain and to the percentage of patients with complete relief of pain. No concomitant factor (including ulcer size, ulcer location, smoking history, or regular alcohol use) affected healing rates in this study. Famotidine was well-tolerated and no serious clinical or laboratory adverse effects were judged to be related to this dosing regimen of famotidine. In conclusion, suppression of nocturnal acid secretion with famotidine (40 mg at bedtime) was more effective than placebo in promoting the healing of acute benign gastric ulcer and its associated symptoms. The results of this study suggest that suppression of nocturnal acid secretion alone is as effective as "around the clock" acid suppression in the healing of benign gastric ulcer. PMID- 2570732 TI - Use of microbleeding and an ultrathin endoscope to assess gastric mucosal protection by famotidine. AB - We have developed an ultrathin endoscope for repeated endoscopy in unsedated subjects and used it with assessment of bleeding rates to investigate aspirin induced gastric mucosal injury and its prevention by famotidine. Compared with placebo, 900 mg of aspirin b.i.d. taken for 48 h caused significant endoscopic injury (median grade 3.5, interquartile range 2-4, modified Lanza scale, p less than 0.01), with an increase in mucosal bleeding from 2.0 (geometric mean; 95% confidence limits, 1.1-3.9) microliters/12 min, to 8.3 (2.4-28.8) microliters/12 min (p less than 0.05). Famotidine (20 mg b.i.d.) raised intragastric pH and reduced endoscopic antral injury (median 1.5, interquartile range 0.5-2, p less than 0.05) and bleeding [3.1 (1.2-8.3) microliters/12 min, p less than 0.01] to levels not significantly different from placebo [1 (0-1) and 2.0 (1.1-3.9) microliters/12 min, respectively]. By contrast, 2 mg of famotidine b.i.d. had no significant effect on intragastric pH endoscopic injury or bleeding rates. The two assessments of gastric mucosal injury correlated strongly (r = 0.71, p less than 0.01). The reduction in bleeding with famotidine tended to be higher, the greater the intragastric pH (r = 0.66, p = 0.057). Ultrathin endoscopy is a simple technique that validates gastric mucosal bleeding as a measure of acute gastric mucosal injury in humans. Acid suppression is an effective method of ameliorating this injury. PMID- 2570733 TI - Effects of dineopentyl and dipinacoline ethers of glutamic acid on neuromuscular transmission in locust (Locusta migratoria). PMID- 2570734 TI - Definition of an efficient synthetic poly(A) site. AB - We constructed and analyzed a synthetic poly(A) (SPA) site that was based on the highly efficient poly(A) signal of the rabbit beta-globin gene. By use of the SPA, we demonstrate that the minimum sequences required for efficient polyadenylation are the AATAAA sequence and a GT/T-rich sequence with the correct spacing of 22-23 nucleotides between them. When placed downstream of the poly(A) site of the human alpha 2-globin gene, the SPA is used exclusively. We predict that the SPA, with its more extensive GT/T-rich sequence, is a more efficient poly(A) site than alpha-globin. Also, we compared the use of the SPA when it is placed either in the exon 3 or intron 2 of the rabbit beta-globin gene. When in the exonic position, SPA is used 10-fold more than the regular poly(A) site of rabbit beta-globin. In contrast, when it is in the intronic location, no detectable use of SPA is observed; however, the deletion of the donor site of intron 2 reactivates the intronic positioned SPA. These results indicate that the splicing of intron 2 in the rabbit beta-globin gene occurs ahead of polyadenylation and have important implications for termination of transcription. Polyadenylation, although required for termination of transcription, is not sufficient; therefore, additional termination signals for RNA polymerase II must exist. PMID- 2570735 TI - Organization of the yeast URA2 gene: identification of a defective dihydroorotase like domain in the multifunctional carbamoylphosphate synthetase-aspartate transcarbamylase complex. AB - The 6636 bp of the yeast URA2 gene encoding the carbamoylphosphate synthetase aspartate transcarbamylase complex have been sequenced. The protein is organized into four regions, three of which are functional domains as indicated previously by genetic analysis. The fourth domain corresponds to a defective dihydroorotase called DHOase-like. The URA2 gene complex with the same organization as the equivalent genes in higher eukaryotes suggests an evolution from a common ancestral gene. PMID- 2570737 TI - DNA typing and forensic science. AB - With new typing techniques forensic scientists can characterise individuals at the fundamental level of their DNA. Variation between individuals at this level can be used to discriminate between them. Why this is so and how it is achieved is described in relatively simple terms for legal and medical practitioners who have no formal training in genetics. PMID- 2570736 TI - Selective sensitization of chemiluminescence resulted from lipid and oxygen radical reactions. AB - Eu3+-tetracycline complex (EuT) increased the chemiluminescence (CL) intensity of linolenic acid micells (UFA-somes) oxidized with lipoxygenase and CL of the lecithin liposomes peroxidized with Fe2+ ions by 3 orders of magnitude. In the systems producing oxygen radicals (xanthine + xanthine oxidase and Fenton's reagent) EuT was ineffective. Luminol increased CL intensity up to 4 orders of magnitude in Fenton's reagent and by 2 orders of magnitude in xanthine oxidase reaction. The sensitization of CL in Fe2+-induced lipid peroxidation (LPO) of liposomes was by a factor 40, while in lipoxygenase reaction very low sensitization was observed. By means of cut-off light filter OS-12 (Soviet) having short wave-length transmittance limit at 560 nm it was possible to measure separately in the same sample the luminol-sensitized CL (maximal emission near 480 nm) and EuT-sensitized CL (maximum at 620 nm); these two CL components reflect, correspondingly, the production rate of oxygen- and lipid-free radicals. Mannitol, the OH radical scavenger, inhibited luminol-dependent component of CL in peroxidized liposomes and did not inhibited EuT sensitized CL in the same system. Apparently, hydroxyl radicals are produced in LPO reactions and responsible for the effect of CL sensitization by luminol, but are not involved in the chain LPO process. PMID- 2570738 TI - Effects of experimental hypo- and hyperthyroidism on hepatic long-chain fatty acyl-CoA synthetase and hydrolase. AB - The effects of T3 treatment and thyroidectomy on rat liver microsomal long-chain fatty acyl-CoA (LCFA-CoA) synthetase and LCFA-CoA hydrolase activities were determined. Hyperthyroid rats had a 36-42% decrease in LCFA-CoA synthetase with no change in hydrolase activity. This may contribute to the redirection of fatty acids from esterification to oxidation reactions in hyperthyroidism. Thyroidectomized rats had a 40-44% decrease in synthetase and a 27-42% decrease in LCFA-CoA hydrolase activity. The decrease in both LCFA-CoA synthetase and hydrolase activities in hypothyroidism may indicate that the LCFA-CoA turnover in this futile cycle is decreased in the liver. PMID- 2570739 TI - Effects of bestatin on intrauterine growth of rat fetuses. AB - Pregnant Wistar rats were injected with bestatin, a specific inhibitor of aminopeptidase M. Placental aminopeptidase M activity was inhibited by injection of bestatin, and fetal body weight was statistically lower than that in the saline-injected or control group. Our present data suggest that placental aminopeptidase M plays an important role in fetal growth. PMID- 2570740 TI - Defective immunoregulation in primary biliary cirrhosis: CD4+, Leu-8+ T cells have abnormal activation and suppressor function in vitro. AB - To determine whether abnormalities of lymphocyte function in primary biliary cirrhosis are due to altered function of immunoregulatory T cell subpopulations, phenotypic and functional characteristics of CD4+ T cells were examined. The proportion of CD4+ T cells expressing the Leu-8 and CD45R antigens was normal in patients with primary biliary cirrhosis. The capacity of CD4+, Leu-8- T cells to provide helper function for pokeweed mitogen-stimulated immunoglobulin synthesis by B cells in vitro was similar in patients and controls. However, in contrast to normal individuals and patients with other liver diseases, CD4+, Leu-8+ T cells from six of 10 patients with primary biliary cirrhosis did not suppress, but enhanced immunoglobulin synthesis. Whereas treatment of CD4+ T cells from normal individuals with anti-Leu-8 monoclonal antibody enhanced their suppressor function, similar treatment of CD4+ T cells from patients with primary biliary cirrhosis did not increase their suppressor function. To determine whether the abnormal regulatory function of CD4+, Leu-8+ T cells was due to a defect of cell activation, the proliferative response of CD4+ T cell subpopulations to mitogenic stimulation was examined. The proliferative responses of CD4+, Leu-8- T cells from patients with primary biliary cirrhosis and controls were similar, but the proliferative responses of CD4+, Leu-8+ T cells from patients with primary biliary cirrhosis were lower than those of control cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570741 TI - Justifying neuroleptic drug treatment. PMID- 2570742 TI - RFLP-discordance within the human phenylalanine hydroxylase locus. AB - A search for crossover events was performed in seven families with two phenylketonuria-affected children applying seven enzymes for the detection of eight restriction fragment length polymorphisms (RFLPs) by hybridisation with a human phenylalanine hydroxylase (PAH) cDNA probe. A discordance was detected within the PvuIIb-RFLP pattern of two affected sibs. This observation is most likely the result of a cross-over event in the PAH locus. Alternative explanations are discussed. Non-paternity could be excluded by DNA fingerprinting using the oligonucleotide probe (GTG)5. PMID- 2570743 TI - Restriction fragment length polymorphism of human aldehyde dehydrogenase 1 and aldehyde dehydrogenase 2 loci. AB - Two probes, ALDH1 and ALDH2, for restriction fragment length polymorphisms (RFLPs) are described, with notes on their locations and the RFLPs found with them. PMID- 2570744 TI - Organization and size class homogeneity of ribosomal RNA genes of catfish Heteropneustes fossilis. AB - The ribosomal RNA genes of catfish Heteropneustes fossilis Bloch were examined by Southern blot analysis of genomic DNA digested with restriction enzymes and probed with labelled catfish ribosomal RNA. The major repeat length is 12 kb and about 300 copies per haploid genome are tandemly arranged. The repeat lengths are homogeneous in size in different tissues and individuals. A restriction site polymorphism exists in some of the repeats. PMID- 2570745 TI - Multiple forms of glutamine synthetase in Bacillus brevis AG 4. AB - Glutamine synthetase in Bacillus brevis AG 4, a Gram-positive spore forming bacteria, has been found to exist in multiple molecular forms. It was purified to electrophoretic homogeneity by single-step Blue Sepharose affinity chromatography. The native enzyme has a molecular weight of 600,000 with subunits of 50,000. The enzyme samples purified from different stages of growth differed in Mg2+ sensitivity and other kinetic properties. Four different enzyme samples selected on the basis of Mg2+ sensitivity showed distinct mobilities at pH 6.3 on PAGE using discontinuous buffer system. A correlation amongst Mg2+ sensitivity, electrophoretic mobility, and kinetic properties was highly suggestive of multiple forms of glutamine synthetase in Bacillus brevis arising due to modification. PMID- 2570746 TI - Modification of glutamine synthetase in Bacillus brevis AG 4. AB - The various forms of glutamine synthetase obtained from Bacillus brevis have been found to be antigenically identical. Alkaline phosphatase treatment of the fast moving form (GS4) reduced the electrophoretic mobility of the enzyme. Radiolabelling and autoradiographic studies have also indicated that 32P incorporation is high in the form depicting high Rm value. Thus, it appears that these forms arise due to covalent modification of the enzyme involving a phosphate group. PMID- 2570747 TI - [Pathogenic neisseriae--model of bacterial virulence and genetic flexibility]. AB - The early stages of an infection with pathogenic Neisseriae are determined by receptor-mediated events that finally lead to the attachment and invasion of human mucous tissues. The factors participating in this process, the pili, OPA proteins, and perhaps lipopolysaccharide (LPS), are subject to complex genetic changes that enable the pathogens to produce multiple variant forms of these factors. Antigenic variation of the pathogenic Neisseriae permits both, the escape from the human immune response and the interaction with different cells and tissues of the human host. One of the intrinsic mechanisms of antigenic variation, i.e. genetic transformation, allows exchange and recombination of virulence genes between independent Neisseria strains in multiply infected individuals. Factors, such as IgA protease, alpha-factor, and the meningococcal capsule are attributed with further striking properties, and thus render the pathogenic Neisseriae as an excellent model for the investigation of bacterial virulence. PMID- 2570748 TI - The RT1.G locus in the rat encodes a Qa/TL-like antigen. AB - A new antigenic system in the rat homologous to the Qa/TL antigen system in the mouse has been characterized. It was detected by antibodies raised in donor recipient combinations that were matched for the RT1.A, B, D, E loci in the major histocompatibility complex (MHC): (R11 X BN)F1 anti-BN.1L(LEW), (R18 X BN)F1 anti BN.1L, and BN.1LV1(F344) anti-BN.1L. Absorption analyses using these antisera and a variety of inbred, congenic, and recombinant strains identified three alleles, RT1.Ga, Gb, Gc, of which Gc is a null allele. The strain distribution of these alleles was determined using 37 strains of rats representative of all of the prototypic haplotypes and a number of congenic and recombinant strains. The use of the congenic and recombinant strains showed that the RT1.G locus was linked to the MHC and that the most probable gene order was A-E-G. Testcross analysis showed that the map distance between A and G was 1.4 cM (4/285 recombinants). The RT1.G antigen has a heavy chain of Mr 46,000 and is present on both T and B cells. PMID- 2570749 TI - Polymorphism in the major histocompatibility complex class II genes of Peromyscus leucopus. PMID- 2570750 TI - Attachment behaviour of immatures of Mansonia annulifera (Theobald), 1901. AB - The detachment and reattachment of the immatures of M. annulifera with their host plants (hydrophytes) is a regular phenomenon. This occurs frequently with the same plant, or with a different one, throughout the larval span. The duration between detachment and reattachment varied from 30 sec to one hour and majority of them get reattached within a minute. This behaviour is instinctive and not found to be influenced by food availability. In the absence of hydrophyte, early immature stages survive relatively longer (192 h), when compared to the late ones. PMID- 2570751 TI - Pellicle receptors for Actinomyces viscosus type 1 fimbriae in vitro. AB - Actinomyces viscosus T14V-J1 and its fimbria-deficient mutant strain possessing type 1 fimbriae strongly aggregated with latex beads treated with acidic proline rich protein 1, basic proline-rich proteins, and proline-rich glycoprotein and its deglycosylated derivative. These type 1+ strains did not aggregate with latex beads treated with other proteins, such as salivary amylase, salivary histidine rich polypeptides, laminin, type 1 collagen, fibronectin, or C1q. The type 1+ strains also adsorbed well to experimental pellicles formed with acidic proline rich protein 1, basic proline-rich proteins, and proline-rich glycoprotein and its deglycosylated derivative on hydroxyapatite (HA) surfaces. These interactions were inhibited with immunoglobulins and Fabs specific for type 1 fimbriae. Type 1 actinomyces exhibited feeble adsorption to latex beads or HA treated with any of the aforementioned proteins. Collectively, these data indicate that actinomyces type 1 fimbriae may specifically interact with several proline-rich salivary molecules, forming experimental pellicles on HA or polystyrene surfaces. PMID- 2570752 TI - Type 1 pili are not necessary for colonization of the streptomycin-treated mouse large intestine by type 1-piliated Escherichia coli F-18 and E. coli K-12. AB - Escherichia coli F-18, an excellent colonizer of the streptomycin-treated mouse large intestine, produces type 1 pili. E. coli F-18 FimA-, type 1 pilus negative, and E. coli F-18 FimH-, type 1 pilus positive but adhesin negative, were constructed by bacteriophage P1 transduction of defective fimA and fimH genes from the E. coli K-12 strains ORN151 and ORN133, respectively, into E. coli F-18. Adhesion of E. coli F-18 to an immobilized mannose-bovine serum albumin glycoconjugate was about sixfold greater than that of either E. coli F-18 FimA- or E. coli F-18 FimH-, and adhesion of E. coli F-18 to immobilized cecal epithelial cell brush border membranes was between two- and threefold greater than that of E. coli F-18 FimA- or E. coli F-18 FimH-. When either E. coli F-18 FimA- or E. coli FimH- was fed to streptomycin-treated mice together with E. coli F-18, the pilus-negative and adhesin-negative strains colonized as well as their type 1-piliated parent. Essentially the same result was observed when the type 1 piliated E. coli K-12 strain ORN152 was fed to streptomycin-treated mice together with a nearly isogenic K-12 FimA- strain, ORN151. Furthermore, when streptomycin treated mice were fed E. coli F-18 FimA- or E. coli F-18 FimH- together with E. coli F-18 Col-, which also makes type 1 pili but is a poor colonizer relative to E. coli F-18 because it grows poorly in mucus in the presence of E. coli F-18, the F-18 FimA- and F-18 FimH- strains colonized well (10(6) to 10(7) CFU/g of feces), whereas the number of E. coli F-18 Col- in feces decreased rapidly to 10(2) CFU/g of feces. These data show that in streptomycin-treated mice, the inability to produce functional type 1 pili has no effect on the ability of E. coli F-18 and E. coli K-12 to colonize the large intestine. PMID- 2570753 TI - Immunological responsiveness of chinchillas to outer membrane and isolated fimbrial proteins of nontypeable Haemophilus influenzae. AB - Thin, nonhemagglutinating fimbriae have been demonstrated on 100% of the clinical isolates of nontypeable Haemophilus influenzae recovered from children with chronic otitis media tested in this laboratory (L. O. Bakaletz, B.M. Tallan, T.M. Hoepf, T.F. DeMaria, H.G. Birck, and D.J. Lim, Infect. Immun. 56:331-335, 1988). Chinchillas with induced otitis media responded to this surface-located antigen of both the infecting and a heterologous strain. Antibodies were found in both serum and middle ear fluids. PMID- 2570755 TI - Relationship of Haemophilus influenzae type b pilus structure and adherence to human erythrocytes. AB - Six strains of Haemophilus influenzae type b, some expressing immunologically different pili, showed identical patterns of binding to erythrocytes that were characterized for 38 blood group antigens. All six strains appeared to bind to the Anton antigen, as they agglutinated all erythrocytes tested except cord erythrocytes and those characterized as Lu(a-b-), dominant type, including Anton negative cells. PMID- 2570754 TI - Efficacy of interleukin-1 beta against systemic Candida albicans infections in normal and immunosuppressed mice. AB - Prophylactic treatments with either recombinant human interleukin-1 beta (rHuIL-1 beta) or a muramyl dipeptide analog ([Abu1]MDP) enhanced the resistance of mice to systemic infection with Candida albicans. The optimum treatment regimen in both normal and cyclophosphamide-treated mice was intraperitoneal administration of 100 ng of rHuIL-1 beta or 1.6 mg of [Abu1]MDP per mouse once daily for 3 consecutive days before infection. Neither rHuIL-1 beta nor [Abu1]MDP was efficacious when started after the infection or when given before cyclophosphamide to mice infected subsequently. Continuing to treat after the infection with either drug neither enhanced nor antagonized the efficacy of prophylactic treatments. PMID- 2570756 TI - Early hematopoietic reconstitution after autologous transplantation with blood derived stem cells in a patient with advanced lymphoma. AB - A 30-year-old man with advanced non-Hodgkin lymphoma underwent repeated leukaphereses for harvesting blood-derived hemopoietic stem cells. Collection was started 8-10 days after the end of L-VAMP therapy (3 cycles). Nine procedures were performed and a total of 65.4 x 10(9) mononuclear cells (0.87 x 10(9)/Kg) were collected, processed, cryopreserved and stored in liquid nitrogen. The yields of CFU-GM, BFU-E and CFU-GEMM were respectively 964 x 10(4) (12.4 x 10(4)/Kg), 249 x 10(4) (3.2 x 10(4)/Kg) and 798 x 10(4) (10.4 x 10(4]. The patient received a myeloblative regimen consisting of fractionated total body irradiation (1200 cGy) and cyclophosphamide (120 mg/kg) followed by infusion of his own thawed cells. Early trilineage hematopoietic recovery was first observed on day +8; 1 x 10(9)/l WBC were reached on day +11, 0.5 x 10(9)/l PMN on day +13 and 50 x 10(9)/l platelets on day +11. Course was uneventful and the patient was discharged from hospital on day +21. Eight months after transplant the patient is in continuous unmaintained complete remission with normal blood cell counts. This reports suggests that complete and sustained engraftment can be achieved with peripheral stem cells recruited after "soft" chemotherapy. PMID- 2570757 TI - Inhibition by cysteamine of hepatocarcinogenesis induced by N-nitrosomorpholine in Sprague-Dawley rats. AB - The effect of cysteamine (2-aminoethanethiol hydrochloride) on hepatocarcinogenesis induced by N-nitrosomorpholine (NNM) was investigated in male Sprague-Dawley rats. Rats received alternate-day s.c. injections of cysteamine, and beginning in experimental week 3 were given drinking water containing NNM for 8 weeks. Pre-neoplastic and neoplastic lesions staining positive for gamma-glutamyl transpeptidase (GGT) or glucose-6-phosphate dehydrogenase (G6PD) were examined by histochemical techniques. In week 18, quantitative histological analysis showed that prolonged administration of cysteamine resulted in a significant reduction in the number of GGT-positive and G6PD-positive hepatic lesions. Histologically, hepatocellular carcinomas were significantly fewer and smaller in GGT-positive and G6PD-positive lesions in rats treated with cysteamine than in untreated rats. Administration of cysteamine also caused a significant decrease in the liver norepinephrine concentration and in the labelling indices of pre-neoplastic lesions and the surrounding liver. Our findings indicate that cysteamine inhibits hepatocarcinogenesis; this may be related to its reducing effect on norepinephrine concentration in the liver and its subsequent inhibition of cell proliferation in neoplastic lesions and surrounding hepatocytes. PMID- 2570758 TI - Terazosin: a new antihypertensive agent with favorable effects on lipids. AB - Terazosin is a new, selective, alpha 1-adrenoceptor antagonist which is structurally similar to prazosin and of similar therapeutic efficacy in the treatment of mild-to-moderate essential hypertension. Unlike prazosin, the terminal-phase plasma half-life is long, at approximately 12 h, and effective blood pressure control is usually achieved with a once-daily dosing regimen. Once daily dosing improves patient compliance, and good compliance to the therapeutic regimen is essential to effective treatment and the reduction of cardiovascular morbidity and mortality. Terazosin has generally been well tolerated in short term and long-term studies. Unlike diuretics and beta-blockers, terazosin does not adversely affect the serum lipid profile. Indeed, beneficial changes in the serum lipid profile have been observed in patients with hypertension. In placebo controlled trials, total serum cholesterol and the combined low-density plus very low-density lipoprotein cholesterol fraction were significantly reduced from baseline in the terazosin-treated group when compared with the placebo-treated group. In addition, terazosin tended to produce increases from baseline in the high-density lipoprotein cholesterol fraction and decreases in serum triglyceride levels. Once-daily dosing and a favorable impact on the serum lipid profile support the use of terazosin as first-choice treatment for mild-to-moderate essential hypertension. PMID- 2570759 TI - Beta-endorphin and dynorphin mimic the circadian immunoenhancing and anti-stress effects of melatonin. AB - We have recently demonstrated that the pineal neurohormone melatonin can enhance immune reactivity in normal mice and counteract the effects of acute stress or corticosterone treatment on antibody production, thymus weight and anti-viral resistance. These remarkable immunopharmacologic effects of melatonin were abolished by naltrexone, suggesting an involvement of the endogenous opioid system. Here we compared the immunopharmacologic action of beta-endorphin, dynorphin 1-13, leu-enkephalin and metenkephalin with that of melatonin in restraint-stressed or prednisolone-treated mice and in normal nonstressed animals. We found that beta-endorphin and dynorphin 1-13 can mimic the immunoenhancing and antistress effect of melatonin. However, at variance with the pineal neurohormone, these opioids were effective in umprimed mcie, too. We found also that restraint stress or prednisolone treatment decreases the immunopharmacologic potency of beta-endorphin and augments that of dynorphin 1 13. In fact, at the doses used, beta-endorphin enhanced the antibody response in normal but not in stressed or prednisolone-treated mice, while dynorphin 1-13 was effective only in counteracting the effect of stress or prednisolone treatment. Most interestingly, all these effects proved to be dependent on the time of administration, i.e. showed a circadian rhythm in analogy with the effects of melatonin. Again, naltrexone abolished all the opioid effects, indicating that their action was exerted via opioid receptors. These findings have important scientific and practical implications. PMID- 2570760 TI - Human T cell lymphotropic virus infection and adult T cell leukemia in the United States. PMID- 2570762 TI - Dissemination of endometrial cells at laparoscopy and chromotubation--a preliminary report. AB - Twenty-one patients undergoing laparoscopy and chromotubation were investigated for the dissemination of endometrial cells into the pelvic cavity. Prechromotubation fluid and postchromotubation fluid from the posterior cul-de sac were aspirated and subjected to cytological assessment. Four patients (15.4%) showed evidence of endometrial cell dissemination into the pelvic cavity. PMID- 2570761 TI - Diethylstilbestrol exposure and the risk of clear cell cervical and vaginal adenocarcinoma. AB - Data from two case-control studies that showed an association between in utero diethylstilbestrol (DES) exposure and clear cell vaginal cancer are evaluated. Neither study considered the cancer risk according to the reasons for prescribing DES and therefore cannot be used to infer any cause-and-effect relationship between DES exposure and vaginal cancer. Neither these studies nor any other studies have specifically evaluated the risk of clear cell cervical cancer to DES exposed women. For DES-exposed women the risk of vaginal and cervical cancer is 0.5-2.0 cases per 1000 DES-exposed women; comparable data are not available for non-DES-exposed women. PMID- 2570763 TI - Immune factor and infertility. AB - Infertility is a common problem seen in gynaecological practice. In a significant number of cases of infertility an immune factor may be operative. In our study of 200 cases, the presence of sperm antibodies was studied in sera of both males and females. The tests used were the gelatin and tube slide agglutination tests. In the infertile group with unexplained infertility (50 couples), 16% of males and 30% of females were positive by the gelatin agglutination test, and 18% of males and 30% of females were positive by the tube slide agglutination test. There was complete absence of positive test (by both methods) in male controls, but females had positive tests for antibodies by both methods in significant numbers. PMID- 2570764 TI - Monthly administration of the LH-RH analogue decapeptyl for long-term treatment of ovarian dysfunctions and estrogen-dependent disorders. AB - The ovarian function of 11 patients suffering from estrogen-dependent disorders and ovarian dysfunction was suppressed by monthly administrations of an analogue of luteinizing hormone-releasing hormone (LH-RH). During the initial 11 to 18 days of treatment, the LH-RH analogue Decapeptyl was subcutaneously injected daily, followed by intramuscular administration of the peptide encapsulated in biodegradable microspheres at 30-day intervals over a 13- to 35-week period. After an initial increase on day 1 of treatment, serum levels of both LH and FSH declined, but did not reach hypogonadotropic values. The pituitary response in the release of both LH and FSH by a LH-RH bolus injection was almost completely suppressed during the treatment course. 17 beta-Estradiol serum levels decreased into the castration range within 9.1 +/- 4 days and remained low in 92% of the values estimated. In one patient, pretreatment mean testosterone levels on the order of 1.9 ng/mL were suppressed to normal within 14 days and remained low under therapy. Seven out of the 11 patients benefited from this therapy. In conclusion, the treatment with Decapeptyl in a slow-release formulation is an effective and suitable approach for the long-term suppression of ovarian function in estrogen-dependent disorders and ovarian dysfunctions. PMID- 2570765 TI - Correlation between plasma estradiol and estrone-3-glucuronide in urine during the monitoring of ovarian induction therapy. AB - An improved radioimmunoassay was developed to determine estrone-3-glucuronide in daily urine. Resulting levels were compared with those of estradiol in plasma of 10 healthy women and 14 undergoing ovulation induction therapy with human menopausal gonadotropin and human chorionic gonadotropin. A highly significant correlation between plasma estradiol and urinary estrone-3-glucuronide in normal (r = .9209; P less than .01) and stimulated (r = .9229; P less than .01) women was demonstrated. These results proved that the pattern of excretion of estrone-3 glucuronide perfectly reflected the changes in plasmatic estradiol levels when monitoring ovarian induction and that estrone-3-glucuronide determinations can provide clinically useful information in human induction therapy. PMID- 2570766 TI - Simplified LH-rise method to time oocyte recovery for in vitro fertilization. AB - The aim of this study was to assess the usefulness of a simplified method to detect the LH rise. The mean plus two standard deviations of the LH values in the prevulatory period was 6.70 +/- 3.45 mIU/mL in a group of women in whom at least one mature oocyte fertilized and cleaved. For practical purposes, an absolute LH value of 15 mIU/mL was considered the start of the rise using the LH graph, and was designated the LH rise value. In 76 patients who experienced a spontaneous LH surge (detected by daily sampling at 08:00, 15:00, 23:00), oocyte recovery was timed for 32 to 35 hours after the LH rise using this simplified method. The outcome of IVF-ET in these patients was comparable to that of the control patients in whom hCG was administered. Graphs were drawn retrospectively using only two (08:00 and 23:00) LH values obtained for each day for each patient and the start of the LH rise was plotted. Most patients (66%) would have had oocyte recovery at the same time whether they had two or three daily LH values. The remaining patients would still have undergone oocyte recovery at an acceptable time (within +/- 6 hours). It was concluded that this simple method accurately identifies the onset of the spontaneous LH rise and may be effected by twice daily LH sampling. PMID- 2570767 TI - The sexual revolution, AIDS and the gynecologist. PMID- 2570768 TI - Leiomyoma of the bladder: case report and review of the literature. AB - Primary mesothelial tumors of the bladder are rare. Although less than 5% of all bladder tumors are malignant in origin, the most common bladder tumor is the benign leiomyoma. Genitourinary leiomyomata may arise in any anatomic structure containing smooth muscle. They have been reported to involve single or multiple organs. Since they may also mimic malignant lesions, they should always be considered in the differential diagnosis of any pelvic mass. Sometimes, depending upon location, such lesions can only be diagnosed after surgical removal. PMID- 2570769 TI - The biological control of homosexual behavior: an essay for modern times. PMID- 2570770 TI - Panel discussion on AIDS. PMID- 2570771 TI - Synthesis and biological studies of dermorphin and its analogs substituted at positions 5 and 7. AB - Dermorphin and seven of its analogs substituted at positions 5 and/or 7, have been synthesized by the solid phase method employing mainly 9 fluorenylmethyloxycarbonylamino acid trichlorophenyl esters in presence of l hydroxybenzotriazole, the solid support being the Merrifield resin. Among the analogs synthesized, the most interesting is [Tyr7]dermorphin. It is one of the most potent dermorphin analogs reported so far. Compared to the natural peptide, it is about two times more potent in the GPI (in vitro) and nearly 1.4 times more potent in its analgesic activity in mice by the hot plate test (in vivo). Further, its antidiarrhoeal activity in mice (in vivo) is comparable to that of dermorphin. On the other hand, [Thr7]dermorphin is almost as potent as dermorphin. PMID- 2570772 TI - Extrinsic denervation elevates neuronal aromatic L-amino acid decarboxylase immunoreactivity in rat small intestine. AB - In order to clarify further the neural control of digestive tract function, we have compared the neuronal localization of tyrosine hydroxylase (TH) and aromatic amino acid decarboxylase (AADC) in rat small intestine. Immunoreactivity for TH was found in numerous varicose axons associated with neurons of the enteric plexuses and in axons within the circular muscular coat and the mucosal villi. Axons with AADC immunoreactivity had a similar distribution, but were sparser in the enteric plexuses and musculature than those containing TH. Chronic extrinsic denervation of a segment of intestine removed all TH-positive nerves from that region. By contrast, the intensity of AADC immunoreactivity was enhanced and more AADC-positive axons were visible than in adjacent intact areas of intestine. The AADC-positive axons appear to represent the intrinsic 'amine-handling' neurons rather than intrinsic tryptaminergic neurons or extrinsic dopaminergic neurons, and the effect on AADC activity of removing the extrinsic nerve supply suggests that this normally exerts some restraining influence on the metabolism of the 'amine-handling' population. PMID- 2570773 TI - Localization of bombesin-, neuropeptide Y-, enkephalin- and tyrosine hydroxylase like immunoreactivities in rat coeliac-superior mesenteric ganglion. AB - The localization of bombesin- (BOMB) and enkephalin- (ENK) immunoreactive (IR) nerves was studied in rat coeliac-superior mesenteric ganglion complex in relation to neuropeptide Y (NPY)- and tyrosine hydroxylase (TH)-immunoreactive neurons with an immunofluorescence double-staining method. Very dense networks of BOMB-IR nerve terminals surrounded the majority of the principal ganglion cells, whether or not they were TH-IR. BOMB-IR nerves were specifically related to the non-NPY-IR neurons. Moderately dense networks of ENK-IR fibers were unevenly distributed among the ganglion cells. Majority of these neurons exhibited TH-IR and some of them also contained NPY-IR. In sections double stained with antibodies to ENK and BOMB some nerve fibers contained both peptides. The findings suggest that BOMB-IR nerves, which have been previously demonstrated to originate from gut, control the function of non-NPY-IR ganglion cells. ENK-IR nerves apparently control the adrenergic neurons which project to gut and also some NPY-IR vasomotoric neurons. The finding that ENK- and BOMB-IR coexist in some nerves suggests that some ENK-IR nerves may originate from gut, although the major part probably represents preganglionic fibers originating from spinal cord. PMID- 2570775 TI - Antihistaminic drugs: H1-receptor antagonists in dogs and cats. PMID- 2570774 TI - Immunohistochemical demonstration of neurotensin and tyrosine hydroxylase in iris nerves of the rabbit eye. AB - In previous studies we have provided evidence that intracameral administration of neurotensin (NT), an endogenous tridecapeptide, produces strong miosis in the rabbit. The presence of NT immunoreactivity was investigated in rabbit iris whole mounts by light microscopic immunohistochemistry, and its distribution in the iris compared to that, of tyrosine hydroxylase (TH). A few scattered NT-positive cell bodies were localized in the dilator muscle. Both, the NT cell bodies and processes appeared parallel to the muscle cells. Extensive branching of NT containing cell processes was observed in connection with the sphincter muscle. These NT-positive fibers formed a dense, randomly oriented network throughout the sphincter muscle cells. The distribution of TH immunoreactivity was similar to that of NT-positive cell processes, except that no TH-positive cell bodies were detected in any of the iris structures examined. Moderate branching of TH positive fibers was observed in the dilator and sphincter iris muscles. These findings provide neuroanatomical support for an important role of NT in pupillary physiology. Its similar topographical distribution with TH suggests that NT and dopamine may be co-localized, as it has already been described in brain. PMID- 2570776 TI - Detection of cells with multidrug-resistant phenotype in myeloproliferative disorders before therapy. AB - The expression of the multidrug resistance (mdr) phenotype is connected with the overexpression of the P-glycoprotein. By applying the immunocytochemical assay, we have demonstrated that in myeloproliferative diseases (AML, ALL, MDS, CGL) in single cases in smear preparations from the peripheral blood as well as from the bone marrow P-glycoprotein-positive cells, respectively, cells with mdr-positive phenotype can be detected in the material obtained from patients before therapy and without clinically and anamnestically known exposure to cytotoxic or immunosuppressive drugs. In the control group of probands without hematologic disorders and also without clinically or anamnestically confirmed contact with cytotoxic or immunosuppressive drugs, we have found P-glycoprotein-positive subpopulations of cells with positive mdr phenotype in a few cases as well. The uniqueness of our results lies in the fact that this finding demonstrates the presence of subpopulations of mdr-positive cells in leukemias and myelodysplastic syndromes before therapy, and furthermore makes evident that a positive mdr phenotype is not necessarily associated with a malignant phenotype or a malignant cell transformation. PMID- 2570777 TI - Alpha 2-adrenergic receptors and the Na+/H+ exchanger in the intestinal epithelial cell line, HT-29. AB - alpha 2-Adrenergic receptors (alpha 2-AR) are negatively coupled to adenylyl cyclase via the GTP-binding protein Gi. However, inhibition of adenylylcyclase does not account for many effector cell responses to alpha 2-AR agonists, suggesting that the receptor can couple to other signal transduction pathways. One potential pathway may be the stimulation of Na+/H+ exchange elicited by alpha 2-AR activation in renal proximal tubule cells, platelets, and the NG-10815 cell line. To determine whether the various receptor-effector coupling mechanisms operate in a tissue-specific manner, we studied the effect of alpha 2-AR activation on basal and stimulated Na+/H+ exchange in epithelial cells isolated from human colon (HT-29 adenocarcinoma cells). Na+/H+ exchange was measured by quantitation of intracellular hydrogen ion concentration (acetoxymethyl ester 2,7 biscarboxyethyl-5(6)carboxyfluorescein) and 22Na+ uptake. HT-29 cells expressed an amiloride-sensitive Na+/H+ exchanger that was activated by reduction of intracellular pH (pHi) to 6.0 but was quiescent at a physiological pHi. The rapid alkalinization observed after acid loading (0.57 +/- 0.07 pH units/min/10(4) cells) was dependent on external sodium and was blocked by amiloride (Ki approximately 2.1 microM). Although epinephrine and the selective alpha 2-AR agonists clonidine and UK-14304 inhibited forskolin-activated adenylylcyclase, these compounds did not alter basal Na+/H+ exchange. Stimulated Na+/H+ exchange was similarly unaffected by epinephrine. In contrast, stimulated Na+/H+ exchanger activity was completely inhibited by the selective alpha 2-agonists clonidine, UK 14304, and guanabenz. This inhibitory effect was not blocked by the alpha 2-AR antagonist rauwolscine, and it is likely due to a direct interaction with the exchanger molecule itself. Structure/activity studies indicated that the compounds inhibiting exchanger activity possess either an imidazoline or guanidinium moiety. Although these molecules bear structural similarity to amiloride, they did not inhibit the amiloride-sensitive epithelial sodium channel in toad urinary bladder, suggesting that these compounds may be useful as "amiloride-like" ligands selective for the Na+/H+ exchanger. These data indicate that in the HT-29 intestinal cell line, in contrast to observations in other tissues, alpha 2-adrenergic receptors are not coupled to the Na+/H+ exchanger, suggesting that the cell-signaling mechanisms utilized by the alpha 2-AR are tissue specific. PMID- 2570779 TI - Identification of a novel proline-directed serine/threonine protein kinase in rat pheochromocytoma. AB - During investigations of the regulation of tyrosine hydroxylase (TH) by protein phosphorylation, a novel protein kinase activity has been discovered in rat pheochromocytoma. Originally detected as a trace contaminant in preparations of highly purified TH, this novel kinase activity phosphorylated TH at serine 8 in the proline-rich amino-terminal region of the enzyme. This particular site is not phosphorylated by, nor is the amino acid sequence surrounding this site selective for, any of the classical (i.e. well characterized) protein kinases. In this report, we describe the identification, characterization, and partial purification of this novel protein kinase. By utilizing a synthetic peptide corresponding to the amino-terminal region of TH, a selective assay for this protein kinase was developed. The kinase activity utilized ATP and magnesium, although GTP could also be utilized as a phosphate donor. The kinase activity was found to co-purify with TH activity through ammonium sulfate precipitation and DEAE-cellulose chromatography and could be only partially resolved from TH by heparin-agarose affinity chromatography. Substantial kinase activity could be resolved from TH by phosphocellulose chromatography. The novel kinase migrates as a protein with a molecular mass of approximately 45 kDa on gel permeation chromatography as well as sucrose density gradient centrifugation. Studies of site specificity indicate that this Ser/Thr kinase activity appears to be directed by an adjacent (carboxyl-terminal) proline residue, exhibiting a minimal recognition sequence of -X-Ser/Thr-Pro-X-. In addition to TH, this proline directed protein kinase will also phosphorylate synapsin I, histone H1, and glycogen synthase, suggesting that this kinase may have multiple substrates in vivo. Additional findings indicate that the activity of proline-directed protein kinase is increased transiently in PC12 pheochromocytoma cells following treatment with nerve growth factor. Distinctions between this novel kinase and other well characterized protein kinases can be made on the basis of phosphorylation site specificity, chromatographic behavior, and physical characteristics. PMID- 2570778 TI - Alternate overexpression of two P-glycoprotein [corrected] genes is associated with changes in multidrug resistance in a J774.2 cell line. AB - In multidrug-resistant murine J774.2 cells, the mdr1a and mdr1b genes encode the 120- and 125-kDa P-glycoprotein precursors, respectively (Hsu, S. I., Lothstein, L., and Horwitz, S.B. (1989) J. Biol. Chem. 264, 12053-12062). It is shown here that a J774.2 cell line selected for vinblastine resistance (J7.V3) switched from the 125- to 120-kDa precursor when cells that were maintained in 20 nM vinblastine were grown in 40 nM vinblastine for 20 months. The rate of switching was accelerated by growing cells in higher levels of vinblastine. These findings suggest that cells which express mdr1a have a selective growth advantage compared to cells which express mdr1b. Consistent with this hypothesis, the switching event that occurs in cells maintained at 40 nM vinblastine was correlated with 3.5-5-fold higher levels of resistance to vinblastine, taxol, and doxorubicin in the absence of any detectable increase in the amount of immunoreactive P glycoprotein. These findings suggest that P-glycoproteins derived from mdr1a and mdr1b are functionally distinct. PMID- 2570780 TI - Complete amino acid sequence of an immunomodulatory protein, ling zhi-8 (LZ-8). An immunomodulator from a fungus, Ganoderma lucidium, having similarity to immunoglobulin variable regions. AB - The complete amino acid sequence of a novel immunomodulatory protein, ling zhi-8 (LZ-8), isolated from a fungus, Ganoderma lucidium (Kino, K., Yamashita, A., Yamaoka, K., Watanabe, J., Tanaka, S., Ko, K., Shimizu, K., and Tsunoo, H. (1989) J. Biol. Chem. 264, 472-478), was determined by protein sequencing. The polypeptide consists of 110 amino acid residues with an acetylated amino end and has a molecular mass of 12,420 Da including an amino-end blocking group. There is no attachment site for an Asn-linked oligosaccharide chain, consistent with the very low carbohydrate content of LZ-8. These results indicate that the native form of LZ-8 with a molecular mass of 24 kDa is a homodimer of the LZ-8 polypeptide whose sequence is described here. Furthermore, the LZ-8 chain shows considerable similarity to the variable region of immunoglobulin heavy chain both in its sequence and in its predicted secondary structure. The interesting possibility that LZ-8 is related to an ancestral protein of the immunoglobulin superfamily is also discussed. PMID- 2570781 TI - A single amino acid substitution in the beta-adrenergic receptor promotes partial agonist activity from antagonists. AB - The family of G-protein-linked receptors includes many important pharmacological targets, of which the beta-adrenergic receptor is one of the best characterized. A better understanding of those factors that determine whether a ligand functions as an antagonist or as an agonist would facilitate the development of pharmaceutical agents that act at these receptors. Site-directed mutagenesis of the hamster beta 2-adrenergic receptor has implicated the conserved Asp113 residue in the third hydrophobic domain of the receptor in the interaction with cationic amine agonists and antagonists (Strader, C. D., Sigal, I. S., Candelore, M. R., Rands, E., Hill, W. S., and Dixon, R. A. F. (1988) J. Biol. Chem, 263, 10267-10271). We now report that substitution of Asp113 with a glutamic acid residue results in a mutant beta-adrenergic receptor which recognizes several known beta-adrenergic antagonists as partial agonists. This partial agonist activity requires the presence of a carboxylate side chain on the amino acid residue at position 113 and is not observed when an asparagine residue is substituted at this position. These observations support the existence of overlapping binding sites for agonists and antagonists on the beta-adrenergic receptor and demonstrate that genetic engineering of receptors can complement structure-activity studies of ligands in defining the molecular interactions involved in receptor activation. PMID- 2570782 TI - Surgical treatment for Takayasu's arteritis. A long-term follow-up study. AB - Thirty patients with Takayasu's arteritis treated surgically were followed for more than ten years postoperatively, 28 women and 2 men. The average age at the initial operation was 29 years. The clinicopathological finding was classified in four types; the distribution of cases was 6 in Type I, 17 in Type II, 4 in Type III, and 3 in Type IV. Surgical procedures performed were carotid artery reconstruction in 9 patients, thoraco-abdominal aortic bypass in 5, renal artery reconstruction in 10, combined procedures in 2, aneurysmectomy in 2, and other procedures in 2 patients. Twenty-six patients survived; four died during the study period. Twenty-three patients (77%) are well with good function of the reconstructed arteries. Six women had safe deliveries after successful operations. However, anastomotic false aneurysms occurred in 5 patients. The surgical management of Takayasu's arteritis must be distinguished from that of atherosclerotic vascular disease. PMID- 2570783 TI - In vitro effects of taxol on ciliogenesis in quail oviduct. AB - When induced by in vivo oestrogen stimulation, ciliogenesis continues in culture in vitro of quail oviduct implants. Ultrastructure of ciliogenic cells was compared after culture for 24 or 48 h in the presence or absence of 10(-5) M taxol. Taxol, which promotes polymerization and stabilization of microtubules, disturbed ciliogenesis, but formation of basal bodies was unaffected by the drug. Conversely, their migration towards the apical surface seemed to be slowed down or blocked and axonemal doublets polymerized onto the distal end of cytoplasmic basal bodies. They elongated and often constituted a more or less complete axoneme, extending between organelles in various orientations. These axonemes, often abnormal, were not surrounded by a membrane, with the exception of the transitional or neck region between the basal body and axoneme. The formation of membrane in this area resulted from the binding of some vesicles to the anchoring fibres of the basal body. They fused in various numbers, occasionally forming a ring, at the site of the transitional region, and exhibited the characteristics of the ciliary necklace. The association of basal bodies with vesicles or with the plasma membrane appeared to be a necessary signal for in situ polymerization of axonemal doublets. In addition, taxol induced polymerization of numerous microtubules in the cytoplasm, especially in the apical part of the cell and in the Golgi area. This network of microtubules may prevent basal body migration. PMID- 2570784 TI - Ischemic flow threshold for extracellular glutamate increase in cat cortex. AB - Extracellular glutamate (Glu), cerebral blood flow (CBF), and auditory-evoked potentials (AEPs) were measured concurrently using microdialysis and hydrogen clearance in the auditory cortex of anesthetized cats during global ischemia of various severities. A threshold-type relationship was observed between extracellular Glu and CBF: Glu increased at CBF levels below about 20 ml/100 g/min. The Glu increase was related to the impairment of AEPs. The results suggest that Glu neurotoxicity is an important factor for ischemic neuronal injury even in penumbra. PMID- 2570785 TI - Ischemic damage in hippocampal CA1 is dependent on glutamate release and intact innervation from CA3. AB - The removal of glutamatergic afferents to CA1 by destruction of the CA3 region is known to protect CA1 pyramidal cells against 10 min of transient global ischemia. To investigate further the pathogenetic significance of glutamate, we measured the release of glutamate in intact and CA3-lesioned CA1 hippocampal tissue. In intact CA1 hippocampal tissue, glutamate increased sixfold during ischemia; in the CA3-lesioned CA1 region, however, glutamate only increased 1.4-fold during ischemia. To assess the neurotoxic potential of the ischemia-induced release of glutamate, we injected the same concentration of glutamate into the CA1 region as is released during ischemia in normal, CA3-lesioned, and ischemic CA1 tissue. We found that this particular concentration of glutamate was sufficient to destroy CA1 pyramids in the vicinity of the injection site in intact and CA3-lesioned CA1 tissue when administered during control (non-ischemic) conditions. In contrast, the same amount injected during ischemia in the CA3-lesioned CA1 region destroyed pyramidal cells in a widely distributed zone around the injection site in the CA1 region. It is concluded that the ischemia-induced damage of pyramidal cells in CA1 is dependent on glutamate release and intact innervation from CA3. PMID- 2570786 TI - Effects of bilirubin on cerebral arterial tone in vitro. AB - Hemoglobin and its metabolite, bilirubin, have been shown to be present in high concentrations in CSF following subarachnoid hemorrhage (SAH). Several reports have indicated that hemoglobin is a potent cerebral vasoconstrictor and therefore is considered to be an active principle in the genesis of cerebral vasospasm. The possible role of bilirubin on the genesis of cerebral vasospasm, however, has not been clarified. The effect of bilirubin on cerebral vessel tone was therefore examined using in vitro tissue bath techniques. Bilirubin (10(-5)-3 X 10(-5) M) induced strong constriction of cerebral arteries from the cat, dog, and pig. The vasoconstriction returned to baseline after bilirubin was washed by prewarmed Krebs solution. Vasomotor responses to various vasoactive substances were then examined after bilirubin was washed away (the bilirubin postwash effect). Norepinephrine (NE)-induced but not serotonin- or acetylcholine (ACh)-induced constrictions were significantly potentiated by bilirubin postwash effect. The potentiated NE-induced constriction was attenuated by yohimbine but not by prazosin. This enhanced vasoconstriction was mimicked by clonidine but not by phenylephrine, suggesting that the potentiation of NE-induced constriction by the bilirubin postwash effect was mediated by the alpha 2-adrenoceptor subtype. The bilirubin postwash effect also resulted in blockade of endothelium-dependent vasodilations induced by A-23147, ACh, and ATP without affecting relaxations induced by direct muscle relaxants such as beta-adrenoceptors, papaverine, and sodium nitroprusside. These results indicate that bilirubin induces direct vasoconstriction, potentiates alpha 2-adrenoceptor-mediated vasodilation. These actions of bilirubin may promote an enhanced overall vasoconstriction in vivo. Bilirubin, therefore, may be involved in the genesis of cerebral vasospasm following SAH. PMID- 2570787 TI - Alterations in the N-methyl-D-aspartate receptor complex following focal cerebral ischemia. AB - The functional integrity of the N-methyl-D-aspartate receptor complex following focal cerebral ischemia in the rat has been examined at a time when brain tissue is irreversibly damaged. Twelve hours after unilateral permanent middle cerebral artery occlusion, [3H]-MK-801 binding was not significantly altered in the ischemic cerebral cortex compared to sham-operated animals. Moreover, the enhancement of [3H]MK-801 binding by exogenous glutamate was preserved in an area of the brain that was permanently damaged by the ischemic insult. PMID- 2570788 TI - [Malignant adrenocortical tumor as a part of Wermer's syndrome. Apropos of a case]. AB - The authors report a case of malignant adrenal carcinoma associated with nodular adrenal hyperplasia as part of Wermer (N.E.M.I.) type pathological polyendocrinopathy syndrome. They stress the prognostic importance of the glandular degeneration and the type of adrenal involvement in these "multiple endocrine neoplasia". PMID- 2570789 TI - High-performance liquid chromatographic method for the simultaneous determination of the enantiomers of carvedilol and its O-desmethyl metabolite in human plasma after chiral derivatization. AB - Quantitative methodology for the simultaneous high-performance liquid chromatographic (HPLC) resolution and determination of the enantiomers of carvedilol, a new multiple-action antihypertensive agent exhibiting both vasodilator and beta-blocking activity, and its active metabolite, O desmethylcarvedilol, in human plasma is described. The method involves reversed phase solid-phase extraction of the analytes, followed by derivatization of the extract with the chiral reagent, 2,3,4,6,-tetra-O-acetyl-beta-D-glucopyranosyl isothiocyanate and injection of the resultant diastereoisomers onto a reversed phase HPLC column coupled to a fluorescence detector. Both pairs of diastereoisomers formed are completely resolved within 12 min (resolution for the respective pairs is 2.26 and 3.32) and the baseline is clean and free from extraneous peaks. The assay is linear over the range 0.6-80 ng/ml of human plasma with a lower limit of detection of approximately 100 pg on-column for each of the enantiomers. The method can be adapted for a number of structural analogues of carvedilol and is currently applied in support of preclinical and clinical studies of the drug. PMID- 2570790 TI - Effectiveness and tolerability of long term treatment with cabergoline, a new long-lasting ergoline derivative, in hyperprolactinemic patients. AB - Cabergoline (CAB) is a new oral dopaminergic compound showing a very long-lasting PRL-lowering activity and reported to be well tolerated. The efficacy and tolerability of chronic treatment with CAB in 30 female hyperprolactinemic patients, aged 18-52 yr (6 microadenomas, 3 macroadenomas, and 21 functional hyperprolactinemias), were studied. In a group of 10 patients who received CAB (0.8 mg once weekly or 0.4 mg twice weekly) for 8 weeks PRL levels normalized while on treatment and remained normal (8 patients) or greatly reduced (1 patient) for 1-2 months after discontinuation of the drug. Twenty-six patients underwent chronic treatment (6-12 months) with an initial dose of 0.5 mg once weekly, subsequently increased to 1-2 mg in 10 patients and decreased in the other 2. Due to severe side-effects CAB was discontinued in 3 patients, in 1, 8, and 12 weeks. A significant reduction of PRL levels was already observed after the first week of treatment (mean +/- SEM basal values, 90.1 +/- 13.3 vs. 29.5 +/ 6.3 micrograms/L; P less than 0.001). Twenty-two patients had normal PRL levels in 1-36 weeks (mean, 6 weeks) with 0.5-2 mg CAB. Twenty-two patients resumed regular menses; 2 patients became pregnant after 3-11 months of treatment. Thirteen patients complained of side-effects (nausea, hypotension, headache, gastric pain, dizziness, and weakness) that disappeared with time in 10 of them. The comparison with a previous bromocriptine treatment regimen in 20 patients had shown that the number of patients requiring discontinuation of the latter drug was significantly higher (7 vs. 3 patients; P less than 0.001). However, 2 patients who needed to discontinue CAB were able to tolerate bromocriptine therapy. A computed tomographic scan performed after 12 months of therapy in 7 patients showed a significant reduction (50%) of the adenoma in 5. In conclusion, our results show that CAB is a well tolerated new dopamine agonist with long lasting activity that represents an advance in chronic medical treatment of hyperprolactinemic conditions. PMID- 2570792 TI - Mannose-resistant hemagglutination, enzyme-linked immunosorbent assay, and immune electron microscopy for detection of K99 fimbrial antigen in Escherichia coli from calves. AB - Mannose-resistant hemagglutination (MRHA) was evaluated for identification of Escherichia coli with K99 fimbriae. The sensitivity and specificity of MRHA, relative to the enzyme-linked immunosorbent assay, were 21 and 79%, respectively. Disagreement between the tests may have been due in part to separation of pili from the cells, with resultant enzyme-linked immunosorbent assay-positive, MRHA negative tests. MRHA was not useful as the sole means for the identification of E. coli with K99 fimbriae. PMID- 2570791 TI - Usefulness of trehalose fermentation and L-glutamic acid decarboxylation for identification of biochemically aberrant Providencia stuartii strains. AB - A total of 849 Providencia isolates were collected during a 4-year period when an increased incidence of nosocomial Providencia stuartii infection was noted in urologic wards. Of these isolates, 630 were identified as P. stuartii, 206 were identified as Providencia rettgeri, and 1 was identified as Providencia alcalifaciens. Twelve inositol-positive isolates from 10 patients (10 strains) resembled P. stuartii in fermenting trehalose but resembled P. rettgeri in fermenting D-arabitol or meso-erythritol or both. The latter traits, however, were not stable in all cases. These aberrant strains were identified as P. stuartii on the basis of their O antigens and DNA hybridization experiments. All isolates were tested for L-glutamic acid decarboxylase activity by a qualitative thin-layer chromatography method. All P. stuartii isolates, including the aberrant ones, were trehalose positive and L-glutamic acid decarboxylase negative. None of the P. rettgeri isolates fermented trehalose, while 99.0% of them and the single P. alcalifaciens strain were L-glutamic acid decarboxylase positive. Thus, trehalose fermentation and L-glutamic acid decarboxylation are more useful for separating P. stuartii from P. rettgeri than are D-arabitol and meso-erythritol fermentation. PMID- 2570793 TI - Quantitation of catecholamine neurons in the locus coeruleus in human brains of normal young and older adults and in depression. AB - A quantitative study of the morphology and distribution of norepinephrinergic neurons in the human locus coeruleus (LC) is given for normal young and older adult brain. Norepinephrine (NE)-producing neurons are identified by immunocytochemistry of two NE biosynthetic enzymes, tyrosine hydroxylase (TH) and dopamine-beta-hydroxylase (DBH), visualized by the peroxidase-antiperoxidase and immunogold-silver-staining methods. TH and DBH immunoreactions yield equivalent results. Both immunocytochemical visualization methods allow detailed analysis of neuronal morphology. The neurons of the human LC fall into four classes: large multipolar neurons with round or multiangular somata, large elliptical "bipolar" neurons, small multipolar neurons, and small ovoid "bipolar" neurons. Though most of the neurons contain neuromelanin pigment, some larger neurons lack pigmentation. Dendritic arborization of all neurons is extensive. Computer assisted quantitative measurements of the parameters somatic size, dendritic arbor length, surface area, and volume are given. Somatic areas of LC neurons of all four classes are decreased in older adult brain, but dendritic arborization is equally extensive as in the younger. The rostrocaudal length of the LC is approximately 15 mm, and no age-dependent decrease is observed. Computer-assisted mapping of immunoreactive neurons and three-dimensional reconstruction allow division of the LC into rostral, middle, and caudal parts with characteristic distribution of neurons. Small neurons predominate in all parts, but the relative contribution of larger cells decreases in a rostrocaudal direction. A cell loss of 27-37% occurs in older adult brains and to 55% in the brain of a chronically depressed patient without dementia. Cell loss is highest in the rostral part, lower in the middle, and absent in the caudal part, and more small cells are lost than larger ones. PMID- 2570794 TI - Alterations in catecholamine neurons of the locus coeruleus in senile dementia of the Alzheimer type and in Parkinson's disease with and without dementia and depression. AB - The present study provides qualitative and quantitative investigations of the norepinephrine (NE) neurons in the locus coeruleus (LC) in two neurodegenerative disorders, the senile dementia of the Alzheimer type (SDAT) and Parkinson's disease (PD). The group of PD subjects was subdivided into cases without dementia (P - D), cases with dementia, L-dopa responsive (P + D), and cases with fulminant dementia whose motor disorder symptoms were L-dopa nonresponsive (P + D/L-dopa non-responsive). NE neurons were demonstrated by immunocytochemistry against tyrosine hydroxylase (TH). Quantitations of neuronal parameters and cell numbers and three-dimensional reconstructions of the LC were carried out with a computer assisted system. In SDAT cases, the rostrocaudal LC length (13 +/- 2.2 mm) is shorter than in controls (14.9 +/- 1.4 mm). The four basic LC neuron classes found in the normal human brain (large multipolar, large "bipolar," small multipolar, and small "bipolar" neurons; Chan-Palay and Asan: J. Comp. Neurol. this issue) are recognizable, but many cell somata are swollen and misshapen with fore-shortened, thick, and less branched dendrites. LC neuron numbers are reduced (between -3.5% and -87.5%). Neuron loss is greatest in the rostral part, less in the middle, and least in the caudal part. In PD cases, the rostrocaudal length (12.4 +/- 1.5 mm) is shorter than in SDAT and controls. The neuronal morphology is more severely altered than in SDAT. The basic neuron classes are hardly distinguishable. Most cell bodies are swollen; they frequently contain Lewy bodies; and the dendrites are short and thin with absent or reduced arborizations. Neuron numbers are more reduced than in SDAT (between -26.4% and 94.4%). Alterations are as severe caudally as rostrally in P - D, and P + D/L dopa nonresponsive cases. P + D cases are more severely affected rostrally. The presence of depression in SDAT and Parkinson's patients is accompanied by the greatest loss of LC neurons. On the basis of morphological alterations of the TH immunoreactive neurons, and the degree and topographical distribution of neuron loss, a differentiation is possible between the LC in normal brain and that in SDAT and PD for diagnostic purposes. PMID- 2570796 TI - A case of mosquito hypersensitivity showing abnormality of peripheral monocyte function. AB - We report here an 11-year-old, Japanese girl with an ulcer-forming type of mosquito hypersensitivity. Since the lesions might have arisen from an Arthus type of hypersensitivity, we used a phorbol myristate acetate (PMA)-triggered chemiluminescence method to investigate the function of her peripheral monocytes, which should play an important role in any Arthus reaction in association with polymorphonuclear leucocytes (PMNs). The chemiluminescence of the patient's monocytes was significantly reduced from that of the normal control. To our best knowledge, no such functional abnormality of the monocytes has been seen previously in patients with an ulcer-forming type of mosquito hypersensitivity. PMID- 2570795 TI - Terfenadine inhibits itch and wheal, but not abnormal erythema, in physical urticarias. PMID- 2570797 TI - The effects of antihistamines on electrophysiologic and behavioral measures of cognition. PMID- 2570798 TI - Prolongation of simple and choice reaction times in a double-blind comparison of twice-daily hydroxyzine versus terfenadine. AB - Newer, nonsedating antihistamines provide a therapeutic alternative for the patient with allergy whose work is impaired by the side effects of traditional H1 antihistamines. To assess the differential effect of these antihistamines on reaction times and subjective symptoms, we compared terfenadine, 60 mg twice daily, to hydroxyzine, 25 mg twice daily, in a double-blind, placebo-controlled, crossover study of 16 healthy, asymptomatic adults. Simple reaction time and choice reaction time were measured with a computer-based, eye-hand, reaction-time testing apparatus. Reaction times and symptom scores were assessed 90 minutes after the fourth and tenth doses of each drug. Hydroxyzine, but not terfenadine, significantly prolonged both simple and choice reaction time (p less than or equal to 0.0001). However, decision time, the time to process one bit of spatial information, was not prolonged by either antihistamine. Therefore, hydroxyzine prolonged the interpretation and response to stimuli of the central nervous system without increasing single-bit processing time. Although terfenadine was not different from placebo for any symptom assessed, hydroxyzine produced significant drowsiness (p = 0.001), dry mouth (p = 0.022), and irritability (p = 0.021). During the 5 days of hydroxyzine administration, neither objective nor subjective symptoms demonstrated the development of tolerance. No correlation was found between subjective symptoms and prolongation of reaction times by hydroxyzine, suggesting that side effect symptoms of traditional antihistamines are unreliable predictors of objective performance. Terfenadine provides a promising therapeutic alternative to traditional antihistamines for individuals performing critical tasks. PMID- 2570799 TI - Differential cognitive effects of terfenadine and chlorpheniramine. AB - The H1-histamine antagonist, terfenadine, has been proposed to have reduced cognitive side effects. In the present study, the relative cognitive effects of terfenadine, 60 mg, chlorpheniramine maleate, 8 mg, and placebo were tested with a double-blind, randomized, three-period crossover design in 24 healthy adult subjects. The dependent variable was latency of the P3-evoked potential. The P3 is a cognitively evoked electroencephalographic response that is an objective and sensitive measure of sustained attention and cerebral processing speed. Disease and drug states that adversely affect the central nervous system can slow P3 latency. For example, the centrally active anticholinergic scopolamine slows cognitive processing speed and prolongs P3 latency. P3 latency (millisecond) means (+/- mean standard error) were pretreatment, 310 (+/- 1.7; placebo, 313 (+/ 3); terfenadine, 320 (+/- 3); and chlorpheniramine, 333 (+/- 3). The findings suggest that terfenadine may be particularly advantageous in patients who require alertness and intact cognitive abilities. PMID- 2570800 TI - L-glutamate decarboxylase immunoreactivity in developing sympathetic tissues of the rat. AB - An indirect immunofluorescence method was used to study the appearance and distribution of L-glutamate decarboxylase (GAD), the enzyme synthesizing gamma aminobutyric acid, in developing rat retroperitoneal sympathetic tissues. GAD immunoreactivity was analyzed in correlation with immunoreactivity to the catecholamine-synthesizing enzyme, tyrosine hydroxylase (TH), in the main retroperitoneal paraganglion, adrenal medullae and abdominal sympathetic ganglia. In different abdominal sympathetic tissues TH-immunoreactive cells first appeared on embryonic days 12.5-14.5, while GAD immunoreactivity was first observed in all these tissues in 14.5-day-old embryos (E 14.5). This suggests that the first expression of GAD is not coupled to the onset of catecholamine synthesis. In developing chain ganglia, GAD immunoreactivity was localized prenatally only in cell clusters with bright TH immunoreactivity, suggesting that GAD is expressed only in the cell lineage leading to ganglionic small intensely fluorescent (SIF) cells. The coeliac-superior mesenteric ganglion complex developed from the preaortic sympathetic tissue, starting from E 16.5 embryos, when the cranial, moderately TH-immunoreactive cells of this tissue were seen to form compact cell islets around the branches of the abdominal aorta. The caudal, intensely TH immunoreactive cells of preaortic sympathetic tissue were seen to form the main retroperitoneal paraganglion from day E 15.5. During the prenatal period GAD immunoreactivity in preaortic sympathetic tissue was present caudally only in these paraganglionic cells and cranially in some brightly TH-immunoreactive cells, representing SIF and/or paraganglionic cells. In the adrenal medulla, only some of the TH-immunoreactive cells showed GAD immunoreactivity during early developmental stages. The moderately TH-immunoreactive, noradrenaline synthesizing, cell clusters were seen for the first time in E 16.5 embryos, and they exhibited no GAD immunoreactivity. Thereafter, GAD was expressed only in the intensely TH-immunoreactive, adrenaline-synthesizing, cell clusters. The results of this study indicate that in the developing rat sympathetic tissues GAD is present only in the cell lineages which differentiate into SIF cells of abdominal sympathetic ganglia, preaortic paraganglionic cells and adrenaline cells of the adrenal medulla. PMID- 2570801 TI - Stimulation via the CD3 and CD28 molecules induces responsiveness to IL-4 in CD4+CD29+CD45R- memory T lymphocytes. AB - Although both IL-2 and IL-4 can promote the growth of activated T cells, IL-4 appears to selectively promote the growth of those helper/inducer and cytolytic T cells which have been activated via their CD3/TCR complex. The present study examines the participation of CD28 and certain other T cell-surface molecules in inducing T cell responsiveness to IL-4. Purified small high density T cells were cultured in the absence of accessory cells with various soluble anti-human T cell mAb with or without soluble anti-CD3 mAb and their responsiveness to IL-4 was studied. None of the soluble anti-T cell mAb alone was able to induce T cell proliferation in response to IL-4. A combination of soluble anti-CD3 with anti CD28 mAb but not with mAb directed at the CD2, CD5, CD7, CD11a/CD18, or class I MHC molecules induced T cell proliferation in response to IL-4. Anti-CD2 and anti CD5 mAb enhanced and anti-CD18 mAb inhibited this anti-CD3 + anti-CD28 mAb induced T cell response to IL-4. In addition, anti-CD2 in combination with anti CD3 and anti-CD28 mAb induced modest levels of T cell proliferation even in the absence of exogenous cytokines. IL-1, IL-6, and TNF were each unable to replace either anti-CD3 or anti-CD28 mAb in the induction of T cell responsiveness to IL 4, but both IL-1 and TNF enhanced this response. The anti-CD3 + anti-CD28 mAb induced response to IL-4 was exhibited only by cells within the CD4+CD29+CD45R- memory T subpopulation, and not by CD8+ or CD4+CD45R+ naive T cells. When individually cross-linked with goat anti-mouse IgG antibody immobilized on plastic surface, only anti-CD3 and anti-CD28 mAb were able to induce T cell proliferation. These results indicate that the CD3 and CD28 molecules play a crucial role in inducing T cell responsiveness to IL-4 and that the CD2, CD5, and CD11a/CD18 molecules influence this process. PMID- 2570802 TI - Interdependence of CD4+ T cells and malarial spleen in immunity to Plasmodium vinckei vinckei. Relevance to vaccine development. AB - We studied immunity to the blood stage of the rodent malaria, Plasmodium vinckei vinckei, which is uniformly lethal to mice. BALB/c mice develop solid immunity after two infections and drug cure. The following experiments define the basis of this immunity. Transfer of pooled serum from such immune mice renders very limited protection to BALB/c mice and no protection to athymic nu/nu mice. Moreover, B cell-deficient C3H/HeN mice develop immunity to P. vinckei reinfection in the same manner as immunologically intact mice, an observation made earlier. In vivo depletion of CD4+ T cells in immune mice abrogates their immunity. This loss of immunity could be reversed through reconstitution of in vivo CD4-depleted mice with fractionated B-, CD8-, CD4+ immune spleen cells; however, adoptive transfer of fractionated CD4+ T cells from immune spleen into naive BALB/c or histocompatible BALB/c nude mice does not render recipients immune. In vivo depletion of CD8+ T cells did not influence the parasitemia in nonimmune or immune mice. Splenectomy of immune mice completely reverses their immunity. Repletion of splenectomized mice with their own spleen cells does not reconstitute their immunity. We conclude that some feature of the malaria modified spleen acts in concert with the effector/inducer function of CD4+ T cells to provide protection from P. vinckei. To be consistent with this finding, a malaria vaccine may require a combination of malaria Ag to induce immune CD4+ T cells and an adjuvant or other vaccine vehicle to alter the spleen. PMID- 2570803 TI - Autoimmune syndrome after induction of neonatal tolerance to alloantigens. CD4+ T cells from the tolerant host activate autoreactive F1 B cells. AB - The induction of transplantation tolerance to H-2b alloantigens in BALB/c (H-2d) mice by neonatal injection of (C57BL/6 x BALB/c)F1 spleen cells, produces an autoimmune lupus-like syndrome due to an activation of persisting F1 donor B cells. This syndrome is characterized by hypergammaglobulinaemia, high levels of anti-DNA antibodies, as well as by circulating immune complexes and glomerular deposits of Ig. The role of host T cells in this model was investigated by using athymic BALB/c nu/nu mice as recipients of normal (C57BL/6 x BALB.Igb)F1 spleen cells. In these "tolerized" BALB/c nu/nu mice, there was a persistence of F1 donor B cells but none of the autoimmune features were expressed, conversely to tolerized BALB/c nu/+ littermates. The injection of CD4+CD8- T lymphocytes from adult normal BALB/c mice in 3-wk-old tolerized BALB/c nu/nu mice triggered the appearance of all the autoimmune findings observed in euthymic tolerant mice. The autoantibodies were produced by persisting F1 donor B cells as shown by allotype analysis. More strikingly, a similar triggering of the autoimmune syndrome, including high titers of anti-DNA IgG antibodies and circulating immune complexes, was observed after injection of CD4+CD8- T cells from 2-wk-old tolerant BALB/c mice into "tolerized" BALB/c nu/nu mice. The anti-ssDNA antibodies were shown to bear only the Ighb allotype, indicating their exclusive origin from F1 donor B cells. These results imply that CD4+ T cells from the tolerant mice are necessary for the activation of autoreactive F1 B cells and for the development of the autoimmune syndrome occurring in this model. They also suggest that, although there is a marked depletion of H-2b-specific alloreactive CTL precursors in those neonatally tolerized mice, this state of tolerance can be associated with the persistence of H-2b-specific alloreactive CD4+ cells. PMID- 2570804 TI - Rabbit major histocompatibility complex. III. Multiple class II DR beta genes and restriction fragment length polymorphism of the class II alpha and beta genes. AB - Several class II alpha and beta chain genes of the rabbit MHC have been cloned and classified into three distinct subregions, R-DP, R-DQ and R-DR, based on their homology to the corresponding HLA-DP, -DQ and -DR genes. The organization of the rabbit MHC class II genes has now been studied in greater detail by analysing genomic DNA of an inbred III/J strain and several other RLA-D homozygous rabbits, with DNA probes derived from cloned R-DR beta genes. Eight previously cloned R-DR beta genes were shown to be allelic forms of five R-DR beta loci. Genomic blot analyses of DNA from seven rabbits homozygous for different RLA haplotypes revealed that the germline contains a total of approximately seven class II beta genes, one DQ beta, one DP beta and five DR beta. Extensive allelic polymorphism was identified by RFLP analysis using DQ and DR probes; limited RFLP was observed with DP probes. RFLP analyses allowed us to distinguish two haplotypes which had not been previously distinguished by MLR. Such RFLP analyses will be useful for identifying MHC 'compatible' rabbits for various immunobiological studies, including transplantation. PMID- 2570805 TI - Hydrophobic properties of Escherichia coli causing acute pyelonephritis. AB - Cell surface hydrophobic properties and expression of P-fimbriae were examined in 130 strains of Escherichia coli derived from women (n = 66) and children (n = 64) with acute non-obstructive pyelonephritis and in 170 faecal strains of E. coli from healthy adults (n = 103) and children (n = 67) by use of the salt aggregation test and the P-fimbriae-specific particle agglutination test. The strains of E. coli isolated were aggregated in salt solutions of varying molarity (0.001-1.6 M final concentration). Patients with predisposing medical or urological conditions in the urinary tract were excluded. Pyelonephritic strains of E. coli from the women and children had a higher degree of cell surface hydrophobicity (80 and 98% respectively) than faecal strains from healthy adults and children (57 and 82% respectively, P less than 0.01 and P less than 0.01). Both pyelonephritic and faecal strains of E. coli from the children were more often salt aggregation positive (hydrophobic) than faecal strains of E. coli from healthy adults (P less than 0.01 and P less than 0.001, respectively). Pyelonephritic strains of E. coli from women and children were more often P fimbriated (79 and 84% respectively) than faecal control strains from women and children (15 and 33%, P less than 0.001 and P less than 0.001, respectively) but there was no significant correlation between expression of P-fimbriae and cell surface hydrophobicity. PMID- 2570806 TI - Somatic mosaicism for DNA repair capacity in fibroblasts derived from a group A xeroderma pigmentosum patient. AB - A female Japanese xeroderma pigmentosum (XP) patient with severe skin lesions and various neurologic abnormalities was assigned to complementation group A by conventional cell fusion studies. Ultraviolet (UV)-irradiated skin fibroblasts showed a biphasic survival curve, as measured by colony-forming ability. The surviving fraction decreased rapidly up to 2 J/m2 of UV, with a steep slope of D(O) (mean lethal dose) = 0.95 J/m2. At much higher doses it decreased more slowly, with D(O) = 3.5 J/m2. To elucidate the cause of this unique survival response, we isolated a large number of independent clones from single colonies and measured their responses to UV. Of 81 clones analyzed, ten showed a marked resistance to killing by UV, which was only slightly more sensitive than normal cells, and these clones had a rate of unscheduled DNA synthesis (UDS) that was about 45% of normal cells. By contrast, the remaining 71 clones were extremely sensitive to UV, typical of XP group A strains, and had a UDS level 1%-3% of normals. Analysis of restriction fragment length polymorphism using seven polymorphic DNA probes indicated that the UV-resistant clones were derived from the same individual as the UV-sensitive clones. These results clearly demonstrate that this patient's fibroblast cells consist of two types with differing responses to UV, and provide direct evidence of somatic mosaicism for DNA repair capacity in an XP patient. PMID- 2570807 TI - Alterations in HLA-DP and HLA-DQ antigen frequency in patients with dermatitis herpetiformis. AB - Dermatitis herpetiformis (DH) is associated with a markedly increased frequency of the HLA class II antigens DR3 and DQw2. To investigate a possible role of HLA DP (or closely associated genes) in the pathogenesis of DH as well as to confirm the previously described alterations of HLA-DR3 and HLA-DQw2 antigen frequency, we have typed 43 patients with DH for HLA-DP, HLA-DQ, and HLA-DR antigens. All patients with DH had typical clinical and histologic features, as well as granular deposits of IgA at the dermal-epidermal junction by direct immunofluorescence. HLA-DR3 was expressed in 41 of 43 (95%) DH patients, whereas HLA-DQw2 was expressed in all 43 (100%). The overall distribution of HLA-DP antigens in patients with DH was significantly different from that seen in all controls and in HLA-DR3 and HLA-DQw2 controls (p less than 0.02). Examination of the frequency of individual DP antigens revealed that HLA-DPw1 was increased (42% of patients with DH vs 11% of all controls and 26% of DR3 positive controls), but this increase was not statistically greater than that expected due to the disequilibrium linkage of DPw1 with DR3/DQw2. Patients with DH, however, did have a statistically significant decreased frequency of DPw2 (14% of patients vs 31% of all controls and 41% of DR3 positive controls) (pc less than 0.05). Studies of three informative families demonstrated that the DPw2 genes of the DH patients were not present on the haplotype thought to carry a DH susceptibility gene (HLA A1, HLA-B8, HLA-DR3, HLA-DQw2). A role of HLA-DP region genes in the pathogenesis of DH is further suggested by the observation that HLA-DPw1 was expressed in 82% (9 of 11) of DH patients with IgA antibodies against dietary antigens as compared with only 33% (4 of 12) of patients without IgA antibodies. HLA-DP genes or genes closely linked to them may be important in DH either as markers of the disease haplotype or by direct involvement in its pathogenesis. PMID- 2570808 TI - [Evaluation of sialyl SSEA-1 antigen in patients with lung cancer]. AB - Stage-specific embryonic antigen-1 (SLX) is a glycolipid recognized by a monoclonal antibody (FH 6) which is produced by cells immunized with mice teratocarcinoma cell line. Using an SLX (Otsuka) kit from Ostuka Assay Laboratory, we measured the serum level of SLX in 96 patients with lung cancer, 305 patients with non-cancerous lung disease and 86 healthy adults, prospectively. When we adopted a cutoff level of 42U/ml (mean + 2S.D. in the healthy adults), the overall positive rates were 29.2% in the lung cancer patients, 15.1% in the non-cancerous patients and 0% in the healthy adults. The positive rate of the lung cancer patients was significantly higher than in the patients with non-cancerous lung disease (p less than 0.05). According to the histological type of lung cancer, the positive rates were 40.9% in 44 patients with adenocarcinoma, 18.4% in 37 patients with squamous cell carcinoma, 0% in 7 patients with small cell carcinoma, and 0% in 3 patients with large cell carcinoma. Staging of the lung cancer patients revealed positive rates for serum SLX of 5.6% in 18 stage I patients, 22.2% in 9 stage II patients, 33.3% in 21 stage IIIA patients, 33.3% in 27 stage IIIB patients, 42.9% in 21 stage IV patients. After we performed the immunostaining method for SLX using FH6, positive immunoactivity was demonstrated mainly in the cell membrane of adenocarcinoma cells. SLX seems to be a tumor-associated marker in patients with lung adenocarcinoma. PMID- 2570809 TI - The irradiation of V79 mammalian cells by protons with energies below 2 MeV. Part I: Experimental arrangement and measurements of cell survival. AB - The relative biological effectiveness (RBE) has been determined for protons with mean energies of 1.9, 1.15 and 0.76 MeV, from measurements of the survival of V79 Chinese hamster cells. The cells are supported as a monolayer and are swept through a beam of scattered protons produced using a 4 MeV Van de Graaff accelerator. An estimation of the dose and unrestricted linear energy transfer (LET) variation within the sensitive volume of the cells is given for the three proton energies. The RBEs for cell survival (relative to 250 kVp X-rays) at the 10 per cent survival level are 1.6, 1.9 and 3.36 for protons with track-average LETs of 17, 24 and 32 keV microns-1 respectively, and the data suggest that protons are most effective at about 40-50 keV microns-1. It is shown that the proton RBEs can be reconciled with those of other light ions if plotted against z*2/beta 2 (where z* is the effective charge and beta is the relative velocity) rather than against LET. PMID- 2570810 TI - Similarities in the repair kinetics of sublethal and potentially lethal X-ray damage in log phase Chinese hamster V79 cells. AB - We have compared the repair kinetics of X-ray-induced sublethal damage (SLD) and potentially lethal damage (PLD) in V79 cells during postirradiation incubation in growth medium at 37 degrees C. Kinetics of SLD repair were studied by dose fractionation (D1 + D2 = 2 + 2, 4 + 2, 4 + 4, 10 + 4 and 10 + 7 Gy). Kinetics of PLD repair were studied by incubating cells with 0.5 M hypertonic saline (HS) for 20 min at various times after irradiation. The repair of SLD, after 2, 4 and 10 Gy, was complete in about 11, 23 and 62 min, respectively. This repair time was independent of the magnitude of dose D2 (test dose). The repair of PLD, after 2, 4 and 10 Gy, was also complete in about 9, 23 and 61 min, respectively. These data indicate that: (1) the kinetics of repair of SLD for a given dose are independent of the magnitude of the test dose D2; (2) the time taken to complete the repair (repair time) is dependent on dose D1, i.e. the larger the dose D1 the longer the repair time; (3) for a given dose, the repair times assayed either by dose fractionation or by HS treatment are similar; and (4) similarities in the repair times may imply that SLD and PLD are the same. The relationship between the repair kinetics and the shape of the shoulder on fractionated survival curves is discussed. PMID- 2570811 TI - Repair of DNA single- and double-strand breaks in proliferating and quiescent murine tumor cells. AB - We evaluated the relationship between the repair of DNA single- and double-strand breaks and cellular radiosensitivity in proliferating vs. quiescent cells of the mouse mammary tumor lines 66 and 67 in vitro, using the technique of filter elution at pH 12.2, pH 7.2 and pH 9.6. In these lines, quiescent (Q; unfed plateau-phase) cells are more radiosensitive than are proliferating (P) cells. At doses of 4-6 Gy, both 66 and 67 Q cells repair single-strand breaks (ssb) with kinetics similar to those of P cells. However, repair of ssb was slightly retarded in Q cells at a higher dose (10 Gy) than at the lower doses. In contrast, repair of ssb in P cells was dose-independent, at least for doses up to 10 Gy. The rate of repair of DNA double-strand breaks (dsb), measured at pH 7.2, was dose-independent in P and Q cells of both lines. The repair kinetics were biphasic, with an initial half-time less than 15 min, and the early phase was similar in all cell groups. The half-time for repair in the slow phase ranged from about 2 to greater than 20 h. The fraction of damage repaired by the slow phase was relatively high in all cell groups (40-70 per cent). In line 66, P cells repaired a higher percentage of dsb by 2 h postirradiation than did Q cells. The opposite was observed in line 67: Q cells repaired more dsb in 2 h than did P cells. The survival of 66 St4 cells (Q cultures which have been refed with complete medium and incubated 4 h) was significantly greater than that of 66 Q; nevertheless St4 cells repaired both ssb and dsb at rates similar to those of Q cells. Therefore, survival does not necessarily correlate with the rates of either ssb or dsb repair among these cell lines in different growth states. PMID- 2570812 TI - Effect of WR-2721 on radiation-induced lipid peroxidation and enzyme release in erythrocytes and microsomes. AB - The effect of WR-2721 on radiation-induced lipid peroxidation and enzyme release was studied after gamma-irradiation of microsomes and erythrocytes. WR-2721 rendered protection to microsomes by reducing lipid peroxidation whereas it did not exert significant protection against radiation-induced lipid peroxidation in erythrocytes. The non-protective effect of WR-2721 was also observed in radiation induced enzyme (AChE) release of erythrocytes. It is proposed that erythrocytes do not dephosphorylate WR-2721 into the free thiol derivative (WR-1065). This was confirmed by the microsomal treatment of erythrocytes that had shown the protective effect of WR-2721. Further, a significant increase in sulphydryl content in microsomes, and no such increase in erythrocytes, after the WR-2721 treatment indicated that dephosphorylation of WR-2721 is necessary for its protective effect. PMID- 2570813 TI - Is the NAD-poly (ADP-ribose) polymerase system the trigger in radiation-induced death of mouse thymocytes? AB - Agents that induce DNA strand breaks evoke a drop in the NAD content in mouse thymocytes. A decrease in the endogenous NAD content that occurs immediately after gamma-irradiation of thymocytes is entirely attributed to the activation of poly(ADP-ribosylation). The addition of 5 mM benzamide before irradiation prevents the postirradiation drop of the NAD level but has no effect on chromatin degradation and cell death. In contrast to liver nuclei, pre-incubation of mouse thymic nuclei with NAD had no effect on the subsequent chromatin endonucleolysis by Ca2+/Mg2+-dependent endonuclease. It is suggested that the NAD-poly(ADP ribose) polymerase system is probably not the trigger in the radiation-induced programmed death of mouse thymocytes, but may merely be indicative of the radiation response of these cells. PMID- 2570814 TI - The induction of micronuclei in human lymphocytes by in vitro irradiation with alpha particles from plutonium-239. AB - Human peripheral blood lymphocytes were irradiated for 24 h in vitro with plutonium-239 citrate. The dose response for micronucleus induction using the cytochalasin B blocking technique fitted well to a linear relationship. Comparison with published data from the same laboratory using X-irradiation, indicated an RBE of 3.6 for alpha particles at low doses. PMID- 2570815 TI - Effect of tritiated water on female germ cells: mouse oocyte killing and RBE. AB - The relative biological effectiveness (RBE) of tritiated water (HTO) was investigated using mouse immature oocytes. Juvenile mice, having immature oocytes that are highly sensitive to radiation killing, were exposed to HTO, 137Cs gamma rays, 60Co gamma rays, or 252Cf fission neutrons. The 137Cs gamma rays were used to simulate the changing dose rate (but not the radiation quality) received from HTO (see section 2). On the 14th day after birth, female ICR strain mice were injected once in the abdominal cavity with HTO at levels of 1.70, 3.40, 6.81, or 10.21 MBq/10g body weight, and their ovaries were removed 2 weeks after the injection. (The cumulative doses are equivalent to 0.039, 0.077, 0.159 and 0.246 Gy, respectively.) The survival rate of oocytes was determined by light microscopy inspection of serial sections, and the results of the different exposures were compared. The number of surviving oocytes decreased exponentially with increasing dose of HTO or 137Cs, and RBEs of HTO compared to 137Cs gamma rays ranged from 1.1 to 3.5. The relative effectiveness of the different types of radiations was compared, and the results of the comparison, in order of increasing effectiveness, was 137Cs, 60Co, HTO, and 252Cf. The RBE of 252Cf compared to 60Co gamma rays ranged from 1.6 to 3.5. PMID- 2570816 TI - Relationship between cell cycle stage in the fertilized egg of mice and repair capacity for X-ray-induced damage in the sperm. AB - The potentiation effects of 3-aminobenzamide, caffeine, hydroxyurea and arabinofuranosyl cytosine on the yield of X-ray-induced chromosome aberrations of mouse sperm were examined at the first-cleavage metaphase, to clarify a correlation between chromosome aberrations and cell cycle dependency of repair capacity of the fertilized egg. The result provided evidence that there are two major types of DNA damage in X-irradiated sperm: (1) short-lived DNA lesions; the lesions are subject to repair inhibition by agents added in G1 and are converted into chromosome-type aberrations during G1, and (2) long-lived DNA lesions; the lesions persist until S phase and repair of the lesions is inhibited by caffeine, hydroxyurea and arabinofuranosyl cytosine in G2. PMID- 2570817 TI - Toxicity of various combinations of X-rays, caffeine, and mercury in mouse embryos. AB - Preimplantation mouse embryos in vitro were exposed to various doses of X-rays (0.25-2 Gy) and to different concentrations of two chemicals: caffeine (0.5-2 mM) and mercury (0.5-5 microM). X-irradiation was given first, followed immediately by exposure to the chemicals. The effects of the agents, applied either singly or in combination of two or of three, were studied using morphological, proliferative and cytogenetic endpoints (formation of blastocysts, hatching, trophoblast outgrowth, formation of inner cell mass, cell numbers, micro-nucleus frequency). The term 'enhancement in risk' was used whenever the effects observed after combined exposure (two or three agents) significantly exceeded the sum of the effects due to the component individual agents. The enhancement in risk observed after exposure to the three agents could be explained by the interactions already detected at the level of a combined exposure to only two agents. There was no increase in risk specific for the presence of all three agents. PMID- 2570819 TI - Repair kinetics in mouse lung: a fast component at 1.1 Gy per fraction. AB - Four experiments to measure the half-time of repair of radiation damage in mouse lung are described. Three of these experiments used two or four doses per fraction of 3.6-9 Gy. Half-times of approximately 0.6 h were found. The fourth experiment investigated repair between the two doses in pairs of small doses given daily (26F x 1.1 Gy in 13 days). Evidence was obtained of more rapid repair between these small fractions, with a half-time of 0.30 h. Using the best methods of analysis appropriate to each experiment, the 95 per cent confidence limits for the T1/2 values at 1.1 Gy per fraction did not overlap with those for the large doses per fraction. PMID- 2570818 TI - Scheduled and unscheduled DNA synthesis in chick embryo liver following X irradiation and treatment with DNA repair inhibitors in vivo. AB - Three hours following X-irradiation of chick embryos with doses of 4 and 8 Gy the in vitro incorporation of tritiated thymidine [( 3H]dT) into DNA (scheduled DNA synthesis, ss) of hepatocytes was reduced to about one-third. Within 24 h after the exposure, ss returned to control values. The return of ss to a normal rate could be strongly inhibited by 2',3'-dideoxythymidine (ddT), and to a lesser extent by 1-beta-D-arabinofuranosylcytosine (araC). In strong contrast to ss, the hydroxyurea (hu)-resistant [3H]dT incorporation (unscheduled DNA synthesis, us) showed a highly significant increase 24 h after treatment of the embryos with araC and/or X-irradiation. Autoradiographic studies revealed no change of total [3H]dT labelling frequency in the whole chick embryo liver 24 h after treatment with araC and/or X-irradiation, but a persistent depression of ss and a simultaneous increase of us. The histological discrimination between affected and non-affected areas argue for a stimulation of DNA synthesis as an antecedent of subsequent mitosis and reparative proliferation adjacent to cell necrosis. It is suggested that the fast recovery of ss in the 12-15-day-old chick embryo is due to an efficient DNA repair system for which DNA polymerase beta is important. The increase of hu-resistance may be an expression of an aberrant DNA synthesis. PMID- 2570820 TI - Modification of tumour response by calcium antagonists in the SCVII/St tumour implanted at two different sites. AB - Dose responses for the effects of four calcium antagonists on SCCVII/St back and leg tumour sensitivity to 20 Gy X-rays, and on tumour perfusion indicated by relative uptake of 86rubidium, were determined. The effects of these agents on 86rubidium uptake in some normal tissues were also investigated. Flunarizine sensitized tumours to X-rays over the dose range 0.005-500 mg/kg given i.p., but only small increases in tumour perfusion were seen at doses of 0.05-5 mg/kg. Verapamil increased tumour radioresistance at doses of 20 mg/kg and above, and reduced tumour perfusion at 50 mg/kg. Below 10 mg/kg, verapamil sensitized the tumours to X-rays, but produced little or no increase in tumour perfusion. Nifedipine at 10 mg/kg and above produced very radioresistant tumours, with correspondingly large reductions in tumour perfusion. At doses below 0.5 mg/kg sensitization was seen, but no increased tumour perfusion. Diltiazem at 50 mg/kg also increased tumour radioresistance with a reduction in tumour perfusion, and at lower doses sensitized tumours to X-rays, with small increases in tumour perfusion. The effects of these agents on normal tissues were varied and difficult to interpret. The similarity between the tumour radiation responses to the four calcium antagonists suggests that they may be acting upon the tumour vasculature in a manner independent of the systemic circulation. PMID- 2570821 TI - Changes in the structure of nucleoids isolated from heat-shocked HeLa cells. AB - Using a technique to detect changes in DNA supercoiling which allows one to visualize both DNA unwinding and rewinding in presence of the intercalating dye, propidium iodide (PI), we showed that hyperthermic treatment (30 min at 45 degrees C) of HeLa S3 cells alters the response to the intercalating dye. Depending on the treatment conditions, we observed a reduction in the maximum size of the DNA loop that can be measured at the relaxation point (PI concentration 5-7.5 micrograms/ml). Cellular heating also affected all degrees of DNA rewinding (measured as a function of PI concentrations between 10 and 50 micrograms/ml). By 6 h after cellular heating these heat effects had disappeared. This time interval correlated with the time necessary for recovery from a heat induced increase to normal nuclear and nucleoid protein content. Using gel electrophoresis we showed that the nucleoids (DNA plus nuclear matrix proteins) after heat exposure are enriched in several polypeptides and that there is a specific increase in HSP 72/73. We hypothesize that the altered response to the intercalating dye after cellular heat shock is due to an increase in polypeptides associated with the nuclear matrix thereby altering the DNA-nuclear protein matrix anchor points. PMID- 2570822 TI - Response to: 'Are Solitons Responsible for Energy Transfer in Oriented DNA?'. PMID- 2570823 TI - New CD4(+) cell line susceptible to infection by HIV-1. AB - Infection of a newly described human T lymphoid cell line, CEM-CL10, with three different variants of HIV-1 resulted in cytopathic effects followed by cell lysis. Following primary lytic infection, proviral DNA could not be detected by Southern blot analysis in the outgrowth of the surviving CEM-CL 10 cells 60 days after initial exposure to HIV-1. These surviving cells could be further grown as a separate line, derived from CEM-CL10, and were found to be resistant to subsequent infection by HIV-1. A marked decrease in CD4 antigen expression was observed in these latter cells but not of the CD3 and transferrin receptor antigens. This decline in cell surface CD4 expression was correlated with both an absence of specific CD4 mRNA and with changes in structure of the CD4 gene. Both the HIV-1-sensitive CEM-CL10 cell line and its CD4(-), HIV-1-resistant derivative line, will be made available to interested investigators. PMID- 2570824 TI - A molecular modeling study on binding of drugs to calmodulin. AB - Computer-graphical methods have been used to study the interaction between a series of drugs and calmodulin. Based on the X-ray crystallographic coordinates of the alpha-C atoms of calmodulin, a molecular model of the helical sequences was built. The model has been used to derive two possible binding sites for phenothiazines and one binding site for penfluridol. The principal binding forces occur through contacts between acidic amino acids of calmodulin and the protonated side-chain nitrogen of the drugs as well as between a basic amino acid and the electronegative substituents of the aromatic rings. Calculated interaction energies show a good correlation with experimental inhibition data. PMID- 2570825 TI - Molecular modeling of a putative antagonist binding site on helix III of the beta adrenoceptor. AB - In recent biochemical studies it was demonstrated that residue Asp113 of the beta adrenoceptor (beta-AR) is an indispensable amino acid for the binding of beta-AR antagonists. Earlier fluorescence studies showed that a tryptophan-rich region of the beta-AR is involved in the binding of propranolol, the prototype beta-AR antagonist. Bearing these two biochemical findings in mind, we explored the beta AR part containing Asp113, for an energetically favorable antagonist binding site. This was done by performing molecular docking studies with the antagonist propranolol and a specific beta-AR peptide which included, besides Asp113, two possibly relevant tryptophan residues. In the docking calculations, the propranolol molecule was allowed to vary all its internal torsional angles. The receptor peptide was kept in an alpha-helix conformation, while side chains relevant to ligand binding were flexible to enable optimal adaptations to the ligand's binding conformation. By means of force-field calculations the total energy was minimized, consisting of the intramolecular energies of both ligand and receptor peptide, and the intermolecular energy. We found an antagonist binding site, consisting of amino acids Asp113 and Trp109, which enabled energetically favorable interactions with the receptor-binding groups of propranolol. According to these results, binding involves three main interaction points: (i) a reinforced ionic bond; (ii) a hydrogen bond; and (iii) a hydrophobic/charge transfer interaction. The deduced binding site shows a difference in affinity between the levo- and dextrorotatory isomers of propranolol caused by a difference in ability to form a hydrogen bond, which is in conformity with the experimentally observed stereoselectivity. Moreover, it also provides an explanation for the beta 1-selectivity of p-phenyl substituted phenoxypropanolamines like betaxolol. The p-phenyl substituent of betaxolol was shown to be sterically hindered upon binding to the beta 2-AR peptide, whereas this hindrance is very likely to be much less with the beta 1-AR peptide. Finally, the proposed antagonist binding site is discussed in the light of some recent biochemical findings and theories. PMID- 2570826 TI - Effects of SCH23390 injection into the dorsal striatum and nucleus accumbens on methamphetamine-induced gnawing and hyperlocomotion in rats. AB - The effects of SCH23390, a selective D1 receptor antagonist, injected into either the dorsal striatum or nucleus accumbens on methamphetamine-induced stereotyped gnawing and hyperlocomotion in rats were investigated. SCH23390 injected into the dorsal striatum did not alter the gnawing induced by both methamphetamine and apomorphine. However, SCH23390 injected into the nucleus accumbens significantly reduced methamphetamine-induced gnawing without altering the effects of apomorphine. Injection of SCH23390 into the nucleus accumbens reduced the hyperlocomotion produced by methamphetamine more markedly than injection of SCH23390 into the dorsal striatum. PMID- 2570827 TI - Chronic pancreatitis and pancreas divisum in an infant: diagnosed by endoscopic retrograde cholangiopancreatography and treated with somatostatin analog. AB - We describe an infant with pancreas divisum diagnosed by endoscopic retrograde cholangiopancreatography. This infant had chronic and unremitting pancreatitis that only became amenable to surgery following the use of somatostatin analog after the usual and customary treatment for pancreatitis was ineffective. PMID- 2570828 TI - Immunohistochemical study of neurochemical markers in gingiva obtained from periodontitis-affected sites. AB - Immunohistochemical methods have been used to study the occurrence of neuronal markers in human gingiva from periodontitis-affected sites. In periodontitis affected buccal gingiva densely distributed neurofilament (NF)-immunoreactive (IR) fiber bundles were observed in the deeper parts of the propria, while NF-IR single fibers occurred in the superficial propria and occasionally in the buccal epithelium. Periodontitis-affected gingiva obtained from interproximal sites showed only sparsely distributed NF-IR fibers. Single nerve fibers immuno reactive to the peptides substance P and calcitonin gene-related peptide occurred close to or within the epithelium in both buccal and interproximal gingiva. Around blood vessels neuropeptide Y-, peptide histidine-isoleucine amide- and vasoactive intestinal polypeptide-IR fibers were occasionally observed, while clusters of gamma-melanocyte-stimulating hormone-IR cells were found in the propria, in addition to gamma-melanocyte-stimulating hormone IR nerve fibers. Somatostatin-IR dendritic cells were seen in epithelium and propria of buccal and interproximal gingiva, although a high variability in the number of SOM-IR cells was observed. All neuronal markers studied showed a similar distribution in material obtained from young patients with clinically healthy gingivae, although the number of NF-IR fibers in the propria in these subjects was lower. The results demonstrate that in gingiva obtained from periodontitis-affected sites several different biologically active peptides occur in both nerve fibers and cells. At least some of these substances could possible play a role in the inflammatory process. However, since clinically normal gingiva was shown to contain nerve fibers and cells expressing immunoreactivity to the substances studied, no unique periodontitis-induced expression of the neuronal markers studied was found. Thus, any alteration of these substances during the periodontitis process remains to be elucidated. PMID- 2570829 TI - Preparation of drug-diluent hybrid powders by dry processing. AB - New hybrid powders have been produced by the dry processing of six drugs (oxyphenbutazone, prednisolone, theophylline, indomethacin, phenacetin and aspirin), with potato starch used as a core material, by means of an electric mortar and a powder surface reforming system designed to produce hybrid powders. The hybrid powders obtained immediately after production differed in their structure from interactive mixtures. With the hybrid powders the drug was spread on the surface of the core particle by friction and collision that occurred in the dry process, but with interactive mixtures the drug simply adhered as intact particles to the surface of diluent particles. Scanning electron microscopy and powder X-ray diffractometry indicated that the mechanochemical phenomenon was essential for the production of the hybrid powders. With time, a shape change in the adhering drug was observed as a relaxation process took place, with recrystallization resulting from the release of accumulated energy. The change with time might depend upon the method of producing powders and the physical properties of the drug used, e.g. the smooth layer of indomethacin produced by the powder surface reforming system reverted to fine particles tightly adhering to the starch surface, though no change was observed with prednisolone. PMID- 2570830 TI - Dissolution of theophylline from film-coated slow release mini-tablets in various dissolution media. AB - The dissolution of an experimental formulation of film-coated slow release theophylline mini-tablets has been investigated using the USP paddle apparatus in test media at pH 1.2 (hydrochloric acid), pH 5.4 and 7.4 (phosphate buffers) at 37 degrees C. Monitoring of in-vitro theophylline release over 12 h, under identical hydrodynamic conditions, showed that the dissolution rate at pH 1.2 is substantially greater (95% of total drug content released in less than 10 h) than that in phosphate buffers. The maximum release after 12 h was approximately 20 and 30% of total drug content of the tablet at pH 5.4 and 7.4, respectively. However, in vivo bioavailability after oral administration of tablets to rabbits corresponded to over 95% of total drug, compared with the same dose administered intravenously. The retarded drug release during in-vitro dissolution in phosphate buffer was attributed to a possible interaction of phosphate ions with theophylline molecules at the tablet core-coat interface. These findings indicate that both rate and extent of theophylline release from the slow release coated mini-tablets are highly sensitive to phosphate buffers. The data also emphasize the usefulness of an animal model for assessment of in-vivo drug release and subsequent absorption, during the development of modified release dosage forms. PMID- 2570832 TI - Prediction of the in-vitro human skin permeability of nicorandil from animal data. AB - A method for estimating the in-vitro permeability of human skin to drugs, based on in-vitro permeation studies using animal skins, has been developed. The skins from hairless rats, guinea-pigs, dogs and pigs were used, with nicorandil and deionized water as model drug and solvent in a drug-donor compartment. Diffusion coefficients through the skin barrier, D, and partition coefficients from the drug-donor compartment to skin, K, of the drug, in each species, were calculated by curve-fitting the in-vitro permeation data to a diffusion equation describing the drug permeation through a homogeneous membrane, using a non-linear least squares method. Each barrier thickness, L, was measured microscopically from microtomed skin sections. A positive relationship was found between the skin permeability, Kp, and K value among the four species, but species differences in the D and L values were small in spite of the Kp values being different among the four species. A positive correlation was also observed between the calculated and experimental K values among the four species, and hence it was suggested that the main factor for the species difference in the skin permeability of nicorandil would be the difference in partitioning of the drug from vehicle to stratum corneum. As a result, it has become feasible to predict and estimate skin permeability of nicorandil in humans by substituting each parameter, extrapolated from the animal skin permeation data and partition experiments, in the diffusion equation. PMID- 2570831 TI - Conformational analysis of 9-substituted adenines in relation to their microsomal N1-oxidation. AB - Metabolic N-oxidation of adenine, 9-methyladenine, 9-benzyladenine, 9 benzhydryladenine and 9-trityladenine has been investigated using hepatic microsomes from hamster, guinea-pig, rabbit, mouse, rat, and dog. N1-Oxide formation occurs with 9-benzyladenine and 9-benzhydryladenine using liver preparations of all species examined, although to different extents. The N oxidase activity was found, amongst rodents, in the order hamster greater than mouse greater than rabbit greater than rat greater than guinea-pig. Microsomal preparations from dog liver contained a small quantity of P-450 and yet produced a relatively large amount of the N-oxides, possibly indicating that other cytochromes in addition to P-450 may be involved in the N-oxidation of these compounds. The most favourable conformations of these 9-substituted analogues have been established using computer graphics modelling and 1H NMR techniques. Results obtained confirmed the importance of the stereochemical properties of these compounds in relation to N1-oxidation. These observations substantiate and extend our previous findings on the electronic, lipophilic, and stereochemical factors affecting the N-oxidation of adenine derivatives. PMID- 2570833 TI - Pharmacokinetic profile of idazoxan in the beagle dog. AB - The alpha 2-antagonist idazoxan (2- (2- (1,4-benzodioxanyl))-2-imidazoline) has been given intravenously and orally to five beagle dogs at 1, 3 and 10 mg kg-1 doses. Idazoxan plasma levels were determined by a HPLC method. After intravenous administration, a linear kinetic behaviour was obtained. Half-life and mean residence time values ranged 105.2-117.1 and 138.1-154.0 min, respectively. Total plasma clearance values and volume of distribution at steady state values ranged from 25.6-32.1 (mL kg-1) min-1 and 3.60-4.36 L kg-1, respectively. After oral administration, time to peak values averaged around 1 h. Dose normalized peak concentration values ranged 161-182 ng mL-1. Bioavailability values ranged 60 88%. Low idazoxan bioavailability has been described in other animal species and attributed to a first-pass effect. PMID- 2570834 TI - Interaction of nefopam and orphenadrine with the cytochrome P-450 and the glutathione system in rat liver. AB - Nefopam, a cyclic analogue of orphenadrine, exhibits a type I (substrate) and a type II (ligand) interaction with ferri-cytochrome P-450 in control and phenobarbitone induced rat hepatic microsomes respectively. In-vitro metabolism of nefopam in phenobarbitone-induced microsomes leads to the production of a reactive metabolite which complexes with cytochrome P-450. In contrast to the known complexation of orphenadrine, complexation by nefopam can be inhibited by glutathione (GSH, 0.1-1.0 mM). However, in-vivo administration of nefopam to rats does not diminish the GSH content of liver cytosol nor increase oxidized glutathione levels nor alter the activities of GSH transferase and GSH peroxidase. In-vivo administration does not lead to cytochrome P-450 induction nor cytochrome P-450 complexation as has been shown for orphenadrine. Finally, nefopam inhibits the NADPH dependent endogenous H2O2 production in both control and phenobarbitone-induced microsomes. PMID- 2570836 TI - Specific age-dependence in capacity-limited uptake of propranolol by isolated rat lung. AB - To investigate the effect of age on the pulmonary uptake of propranolol, minced tissue (0.4 g) of lungs isolated from 3- to 104-week-old rats was incubated with the drug (1 to 500 micrograms mL-1) prepared in oxygenated, pH 7.4 Krebs-Ringer bicarbonate buffer solution (20 mL) containing 3% BSA for 60 min at 37 degrees C. In any age-group the metabolism of propranolol was not significant (i.e. less than 0.6% of any initial load) under the present in-vitro conditions. The extent of uptake after the incubation with 2.5 micrograms mL-1 of the drug was largest in the 7 weeks (i.e. 82% of the initial load) and relatively small in the 3(64%), 24(61%), 52(51%) and 104(48%) week old rats. Similar, specific age-dependence was observed in the tissue-to-medium concentration ratio of the drug. In any age group, the initial uptake rate obtained in the first 5 min of the incubation was found to be a combination of apparently linear transport and saturable (capacity limited) processes. There was a marked, specific age-dependence in the capacity limited uptake rate. Although Km' value was almost equivalent in any age-group (i.e. 24.4 to 25.4 micrograms mL-1), Vmax exhibited a specific age-dependence by yielding the highest value in 7 weeks (0.726 +/- 0.101 mg g-1 min-1) and relatively low values in 3 (0.501 +/- 0.082 mg g-1 min-1), 52 (0.410 +/- 0.088 mg 1 g-1 min-1) and 104 (0.397 +/- 0.074 mg g-1 min-1) weeks. PMID- 2570835 TI - Disposition of the hypolipidaemic agent, 2,3-dihydrophthalazine-1,4-dione, in Sprague Dawley rats. AB - The disposition of [14C]2,3-dihydrophthalazine-1,4-dione, a potent hypolipidaemic agent, has been determined after both intravenous and oral administration. Both the routes of administration afforded multi-exponential disposition with an estimated t1/2 of approximately 75 h. After oral administration, the drug was observed to be absorbed rapidly from the intestine and distributed quickly to all tissues of the body. A large quantity of the 14C-radioactivity was found in the skin and carcass. Approximately 35% of the administered radioactivity was excreted in urine after oral administration and 11% in the faeces. Approximately 66% of the radioactivity excreted in urine was the parent drug. There was evidence of an additional metabolite which accounted for 28% of the urinary radioactive excretion. The parent drug has little serum protein binding, is highly water soluble, and is probably taken up by cells by passive diffusion. PMID- 2570837 TI - Human liver and plasma aspirin esterase. AB - The plasma, in addition to the liver, is a major site of hydrolysis of aspirin. Human plasma and liver aspirin esterase activities in samples from a group of patients varied over a two fold range and there was a significant correlation between individual plasma and liver activities. Human liver aspirin esterase was present in the cytosolic and microsomal fractions. Cytosolic and microsomal enzymes had different activities and apparent affinities for aspirin. PMID- 2570839 TI - The effect of Ca deprivation and of Ca-blocking drugs on oxytocin-induced contractions of the male mouse anococcygeus. AB - Oxytocin (4 nM)-induced contractions of the male mouse anococcygeus were rapidly and completely lost in EGTA (2 mM)-containing, Ca-free Krebs solution. Contractions were also lost, although more slowly, in Ca-free Krebs solution without EGTA; under such conditions, readdition of Ca did not by itself cause contraction, but readdition of Ca (0.1-2.5 mM) in the presence of 4 nM oxytocin resulted in a rapid contractile response. These Ca-induced responses, in the presence of oxytocin, and those to oxytocin in normal Ca-containing Krebs solution, were unaffected by nitrendipine (0.01-1 microM). Contractions to oxytocin were completely blocked by the calmodulin antagonists trifluoperazine (50 microM) and W-7 (75 microM). It is concluded that oxytocin-induced contraction of the mouse anococcygeus does not require opening of nitrendipine sensitive Ca channels, and there is no Ca-independent component of the contractile response; the cellular mechanisms linked to the oxytocin receptor in the anococcygeus are therefore different from those in the uterus. PMID- 2570838 TI - Alphaxalone reveals GABAA receptor-mediated inhibition in guinea-pig ileum. AB - Contractions of the transmurally stimulated guinea-pig ileum were not affected by 10 microM bicuculline, a competitive GABAA receptor antagonist. The GABAA potentiator alphaxalone depressed the responses to 2 and 20 Hz stimulation by 20% and 40%, respectively, but had no effect on 0.2 Hz responses. The depressions were prevented by 10 microM bicuculline. This suggests that endogenous activation of GABAA receptors during transmural stimulation was normally subthreshold but, when pharmacologically potentiated, inhibitory effects were seen. PMID- 2570841 TI - Pharmacokinetics of 8-phenyltheophylline in the rat. AB - The pharmacokinetics of the adenosine antagonist 8-phenyltheophylline (8-PT) have been investigated in the rat. After intravenous administration to male rats with normal renal function, 8-PT was rapidly cleared from plasma with a t1/2 of about 14 min. Its apparent volume of distribution was some 76 mL/100 g and plasma clearance was 3.5 mL min-1/100 g. Injection via the carotid artery did not alter the kinetics of 8-PT, but when given through the portal vein a first-pass effect was observed. Moreover, 8-PT could not be detected in plasma following intraperitoneal injection. The kinetics of 8-PT were not altered in male rats with acute renal failure and no difference was noted between male and female animals with respect to the kinetics of 8-PT following intravenous injection. It is concluded that the pharmacokinetic properties of 8-PT are likely to complicate its use in-vivo. PMID- 2570840 TI - Bioavailability of intramuscular vitamin E acetate in rabbits. AB - The bioavailability of alpha-tocopherol acetate and alpha-tocopherol (vitamin E) was assessed in male rabbits given 50 mg kg-1 doses according to a randomized design. After intramuscular injection of alpha-tocopherol acetate in colloidal aqueous solution, a mean absolute bioavailability of 65% was calculated for the acetate and 35% for the physiologically active compound, alpha-tocopherol. Comparison of the kinetic profiles after intravenous and intramuscular administration of the acetate and intravenous injection of alpha-tocopherol, revealed absorption of alpha-tocopherol acetate from the site of injection and hydrolysis of the acetate to be potential limiting steps in the bioavailability of alpha-tocopherol. Intramuscularly injected alpha-tocopherol acetate in olive oil (the only formulation available in a few European countries) proved completely bio-unavailable. It thus appears necessary to re-assess the utility of current vitamin E supplementation, since the only formulations available offer poor bioavailability. PMID- 2570842 TI - Deamination of hordenine by monoamine oxidase and its action on vasa deferentia of the rat. AB - The selectivity of the naturally occurring amine, N,N-dimethyltyramine (hordenine) for monoamine oxidase (MAO) and its action upon isolated vasa deferentia of the rat was investigated. Hordenine was deaminated by rat liver MAO with a Michaelis constant of 479 microM and maximum velocity of 128 nmol (mg protein)-1 h-1 compared with 144 microM and 482 nmol (mg protein)-1 h-1 for tyramine. Studies, with selective irreversible inhibitors of MAO, showed that hordenine was a highly selective substrate for MAO-B of liver and that it was not deaminated by the MAO-A of intestinal epithelium. In contrast to tyramine, hordenine did not produce contractions of isolated vasa deferentia. However, 25 microM hordenine potentiated contractile responses of vasa, from control animals, to submaximal doses of noradrenaline and inhibited responses to tyramine. It did not alter responses, to noradrenaline, of vasa denervated by chronic pretreatment of rats with guanethidine. Therefore, it appears that hordenine acted as an inhibitor of noradrenaline uptake, in isolated vasa deferentia. These results indicate that dietary-hordenine is unlikely to be deaminated by intestinal MAO as this is predominantly MAO-A. Consequently, it is likely to be absorbed and could affect the sympathetic nervous system, by virtue of its action as an inhibitor of noradrenaline uptake. PMID- 2570843 TI - Rapid production of ulcerative disease of the colon in newly-weaned guinea-pigs by degraded carrageenan. AB - In a dose-response study, degraded carrageenan (Eucheuma spinosum) was supplied in the drinking fluid at 1.2 and 3% concentrations over two weeks to young adult guinea-pigs. Ulceration of the large bowel was produced in 100% of animals, the severity and extent of damage probably being dose-related. In a time-course study, 3% degraded carrageenan solution supplied to newly-weaned guinea-pigs produced in 100% of animals ulceration in the caecum by four days and in the ascending colon by seven days. The onset of ulceration occurred as early as the second day. This model is convenient and economic for the screening of drugs of potential therapeutic value in human ulcerative colitis. PMID- 2570844 TI - Hydroxylamine dilates resistance blood vessels of the perfused rat kidney and mesentery. AB - Hydroxylamine (ED50 values, 47 +/- 8.9 nmol and 320 +/- 39 nmol) dilates resistance arterioles of the perfused noradrenaline-preconstricted rat kidney and mesentery. In this respect hydroxylamine was approximately 63x and 320x less potent than acetylcholine (ACh) and 15x and 128x less potent than nitroprusside in the two perfused organs studied. The vasodilator effect of hydroxylamine (unlike that of ACh) was unaffected by CHAPS de-endothelialization suggesting that its effect is independent of endothelium-derived relaxing factor (EDRF). PMID- 2570845 TI - Iontophoretic transport of weak electrolytes through the excised human stratum corneum. AB - The effect of iontophoresis on the rate of permeation of a number of therapeutically active weak acids and bases through excised human stratum corneum has been examined, over a range of pH values. It has been shown that the amount of ionized drug species present in the drug solution is an important factor in the delivery of acids and bases by this route and that the molecular weight of the drug does not influence the rate of delivery by iontophoresis. PMID- 2570846 TI - Biodegradation rate of embolized protein microspheres in lung, liver and kidney of rats. AB - The targeting and sustained release characteristics of cytotoxic drug-loaded protein microspheres may prove useful in the therapeutic chemoembolization of solid tumours. Because biodegradation rate of embolized particles will influence rate of incorporated drug release and duration of exposure, this parameter was studied for microspheres (10-30 microns mean diam.) prepared from the proteins albumin and casein, that we have previously used as carriers for doxorubicin. As a measure of microsphere loss in-vivo the radionuclide 125I was chosen because it can be covalently bound to proteins and also homogeneously distributed throughout the matrix. Radiolabelled microspheres were administered to rats both intravenously (lung as target organ, 1.4-2.2 mg/100 g) and via the hepatic artery (liver as target organ, 0.4-0.8 mg/100 g). In both cases it was observed that the casein system biodegraded more slowly than the albumin in-vivo. Thus, time taken for loss of 50% of embolized microspheres from lung was: albumin 2.0 days; casein 3.5 days and from liver:albumin 3.6 days; casein 6.8 days. Microsphere "debris" did not markedly accumulate in other organs. In-vitro experiments showed that microspheres were stable in serum and that albumin microspheres were not innately more sensitive to enzymic digestion than casein. The results may be useful in estimating duration of exposure of target organs to drug-loaded microsphere systems prepared from these proteins. PMID- 2570847 TI - Controlled release of LHRH agonist, leuprolide acetate, from microcapsules: serum drug level profiles and pharmacological effects in animals. AB - The pharmacokinetic behaviour of leuprolide acetate from a controlled release parenteral dosage form has been studied in rats and dogs. The release of the drug in rats after a single subcutaneous injection exhibited pseudo-zero-order kinetics for one month in doses ranging from 0.0135 to 1.35 mg/rat; the release rate at a dose of 1.35 mg/rat was 2.8% of dose/day; after intramuscular injection the response was similar. In rats, the serum leuprolide acetate levels increased sharply immediately after injection by either route as a consequence of the initial release of the drug; subsequently, the levels attained a plateau for two weeks. The serum level profiles in dogs showed essentially the same pattern as those in rats. When the dosage form was injected into rats, the serum testosterone level (a pharmacological index) sharply peaked, abruptly decreased to below the normal level, and then was sustained at a suppressed level for over six weeks at a dose of 1.35 mg/rat (equivalent to 3 mg kg-1) and higher, while the serum testosterone level after an injection of 0.0135 and 0.135 mg/rat was not sufficiently suppressed. The profiles in dogs showed essentially the same pattern as those in rats. With multiple administrations (once every 4 weeks), serum testosterone levels in dogs did not show any sharp rise after the second and third injections. Changes in rat reproductive organ weights agreed well with the serum testosterone profile in the suppression. The results demonstrate that this dosage form releases the drug at a constant rate for one month and has a long-acting potency. PMID- 2570848 TI - Study on transport of disopyramide into the intestinal lumen aimed at gastrointestinal dialysis by activated charcoal in rats. AB - The characteristics of exsorption and/or excretion of disopyramide into the gastrointestinal lumen have been investigated after intravenous administration of the drug at doses of 10 and 30 mg kg-1 to rats by the in-situ single pass perfusion technique. Disopyramide was appreciably excreted into the bile where its levels were approximately ten-fold higher than those in the serum. The exsorption rate of disopyramide and mono-N-dealkyldisopyramide (MND) into the perfusate was increased with an increase in the serum level following an increase from 10 to 30 mg kg-1 in the dose of disopyramide. The average amounts of disopyramide exsorbed into the perfusate were 17.0 and 18.4% at the dose of 10 and 30 mg kg-1, respectively, whereas those of MND were less than 1% at both doses of disopyramide. Oral administration of activated charcoal reduced the serum disopyramide levels after intravenous administration of the drug (20 mg kg 1) compared with the control treatment. By oral administration of activated charcoal, t 1/2 and AUC were decreased to 89 and 82%, respectively, and Cltot was increased to 122% compared with the corresponding control treatment. Vd was not different between the treated rats and control rats. These results suggest that the oral administration of activated charcoal can enhance the clearance of disopyramide and MND from the blood. PMID- 2570849 TI - Inhibition of Na+,K+-ATPase activity by phospholipase A2 and several lysophospholipids: possible role of phospholipase A2 in noradrenaline release from cerebral cortical synaptosomes. AB - p-Bromophenacyl bromide (PBPB), quinacrine and indomethacin, which inhibit phospholipase A2 (PLA2; EC 3.1.1.4) activity in several tissues, caused a dose dependent inhibition of prelabelled [3H]noradrenaline ([3H]NA) release evoked by high concentrations of K+ from rat cerebral cortical synaptosomes. Release of prelabelled [3H]NA was caused by natural lysophosphatidic acid (LPA; 10(-6)-10( 5) g mL-1) and lysophosphatidylcholine (LPC; 10(-6)-10(-5) g mL-1) and synthetic LPA (6 x 10(-6), 2 x 10(-5) M) and LPC (6 x 10(-6), 2 x 10(-5) M), but not by natural lysophosphatidylserine (LPS; 10(-5) g mL-1), lysophosphatidylethanolamine (LPE; 10(-5) g mL-1) and lysophosphatidylinositol (LPI; 10(-5) g mL-1). The release evoked by natural LPA and LPC could be inhibited only marginally by PBPB and quinacrine. Phosphatidic acid (PA)-specific and phosphatidylcholine (PC) specific PLA2 activities from rat cerebral cortical synaptosomes were stimulated in incubation medium containing high concentrations of K+ or calcium ionophore A23187. Low concentrations of PLA2 (10(-6)-10(-8) g mL-1, from bee venom) inhibited the synaptic membrane Na+,K+-ATPase activity in incubation media with intracellular levels of free Ca2+. Several lysophospholipids (LPLs), metabolites of the PLA2 type, also inhibited the synaptic membrane Na+,K+-ATPase activity in a dose-dependent manner. The minimum effective concentrations of natural LPA, LPC, LPS, LPI and LPE were 10(-6), 4.7 x 10(-6), 10(-5), 4.7 x 10(-5) and 4.7 x 10(-5) g mL-1, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570850 TI - Involvement of sialic acid in high-affinity binding of quaternary ammonium compounds by brush border membrane from rat intestine. AB - As one approach to clarify the absorption mechanisms of quaternary ammonium compounds (QACs), their binding characteristics have been studied using brush border membrane vesicles isolated from rat small intestine and liposomes composed of phospholipid and GM3 ganglioside. The binding of propantheline was significantly decreased when the vesicles were pretreated with neuraminidase. Propantheline and methochlorpromazine bound to the liposomes, the binding for the latter drug being significantly greater than that for propantheline. When GM3, isolated from rat small intestine, was incorporated into the liposomes their binding capacity for both drugs increased significantly. It is suggested that the binding of QACs to the lipid layer and sialic acid play a role in the high binding of drugs to the intestinal brush border membrane. Furthermore, a sensitive and reproducible high-performance liquid chromatographic method for sialic acid has been developed. PMID- 2570851 TI - Rilmenidine differs from clonidine in that it lacks histamine-like activity. AB - Studies were carried out to determine whether rilmenidine, a recently introduced antihypertensive agent with a similar mechanism of action of clonidine, possesses histamine-like activity on tissues responding to histamine H2-receptor agonists. In guinea-pig isolated atria, both histamine and clonidine caused concentration dependent positive chronotropic effects which were blocked by the histamine H2 receptor antagonist cimetidine (5 microM); in contrast, rilmenidine produced a concentration-dependent negative chronotropic effect which was not altered by cimetidine. In stilboestrol-treated rat isolated uterus contracted by KCl, histamine and clonidine produces concentration-dependent relaxations which were blocked by cimetidine (5 microM); rilmenidine also produced relaxation, but this was not affected by cimetidine (5 microM). These findings suggest that rilmenidine, unlike clonidine, does not activate histamine H2-receptors in either guinea-pig atria or rat uterus. PMID- 2570853 TI - Protein binding of salicylic and salicyluric acid in serum from malnourished children: the influence of albumin, competitive binding and non-esterified fatty acids. AB - The serum protein binding of salicylic and salicyluric acid has been determined by ultrafiltration in 60 children after administration of oral salicylate. The children were classified according to nutritional status: well-nourished (n = 12), underweight (n = 12), marasmic (n = 17) marasmic-kwashiorkor (n = 7) and kwashiorkor (n = 12). Salicylic acid free fractions were 0.106 +/- 0.026, 0.114 +/- 0.069, 0.141 +/- 0.037, 0.285 +/- 0.279 and 0.438 +/- 0.190 in the five groups, respectively. Salicyluric acid free fractions were 0.184 +/- 0.057, 0.280 +/- 0.282, 0.236 +/- 0.114, 0.484 +/- 0.497 and 0.646 +/- 0.261, respectively. The degree of binding was dependent on serum albumin levels, ligand concentrations and non-esterified fatty acids (NEFA). The NEFA/albumin ratio ranged from 0.05 to 6.6. The fitting of a one-site Scatchard binding model to the collective data was improved when a decrease was allowed for in the number of binding sites in proportion to NEFA concentrations. Salicyluric acid binding could be fitted only when inhibition of the parent compound was included. Binding was not affected by age or sex. The major determinants of salicylate binding in sera from malnourished children have thus been identified. PMID- 2570852 TI - Effect of pyridoxamine on semicarbazide-sensitive amine oxidase activity of rabbit lung and heart. AB - Rabbit lung and heart show clorgyline-resistant benzylamine oxidase activity which is sensitive to semicarbazide (SSAO) and alpha-amino-guanidine. This SSAO activity is inhibited by pyridoxamine with an IC50 of 6.3 x -6 M for lung and of 1.1 x 10(-5) M for heart, the inhibition being non-competitive and only partially reversed by dialysis at 4 degrees C. Semicarbazide, alpha-aminoguanidine and pyridoxamine show a similar time-dependent type of inhibition of rabbit lung and heart SSAO. PMID- 2570854 TI - Assignment of the C-5 configuration of penicilloic acids. AB - Penicilloates prepared by the method of Munro et al (1978) were examined by nuclear magnetic resonance spectroscopy and optical rotation to assign the configuration at C-5. The configuration was 5S for carbenicillin and ticarcillin penicilloates and 5R for the other seven penicilloates examined. The chemical shift and coupling constants of 5-H and 6-H showed consistent differences between the isomers and the optical rotation of the 5R-isomers is consistently more positive than that of the corresponding 5S-isomer. PMID- 2570855 TI - Solute adsorption and concentration-dependent permeability in certain polymer films. AB - Certain parameters (solute adsorption, permeability and swellability) of films of a cationic acrylatemethacrylate copolymer have been determined as functions of the solute (sulphacetamide sodium) concentrations. Both permeability coefficients and the free (unadsorbed fraction of solute increased almost proportionately during increase in solute concentration 1-5%, above 5%, further increase in the fraction of free solute was slight, while the permeability coefficients decreased slightly. The solute also reduced film swellability but the observed concentration-dependent permeability related more to the adsorption phenomenon. PMID- 2570856 TI - Determination of microscopic dissociation constants of 3-hydroxy-alpha (methylamino)methyl-benzenemethanol by a spectral deconvolution method. AB - Microscopic dissociation constants of 3-hydroxy-alpha-(methylamino)methyl benzenemethanol have been calculated from the titration spectrophotomeric data (c = 3.8 x 10(-4) M. Ionic strength = 0.16; buffer system: H3BO3/KOH) by application of a spectral deconvolution method. The results found (pKa = 9.48; pKb = 9.71; pKc = 10.12 and pKd = 9.88) are in good concordance with those obtained from the conventional regression linear method (pKa = 9.45; pKb = 9.77; pKc = 10.14 and pKd = 9.81). PMID- 2570857 TI - A new approach to prostate cancer. AB - Growth of androgen-dependent human prostatic adenocarcinoma implanted in the nude mouse (Honda tumour), is inhibited by 6-methyleneprogesterone. This steroid is a potent inhibitor of both rat and human prostatic 5 alpha-reductase in-vitro. In vivo, at the studied dose level, it reduces metabolic conversion of testosterone to dihydrotestosterone with minimal effects upon circulating LH and testosterone. These data support the hypothesis that dihydrotestosterone and not testosterone is the main trophic androgen of the human prostatic neoplasm. PMID- 2570858 TI - Stimulation of postsynaptic D2- dopamine receptors by B-HT 958 is revealed by co treatment with the D1- receptor agonist SKF 38393. AB - The motor activity of reserpine-treated mice was used to study effects of B-HT 958 (2-amino-6-(p-chlorobenzyl)-4H-5,6,7,8-tetrahydrothiazolo-[5,4-d]- azepine) on postsynaptic dopamine and noradrenaline receptors. The motor activity was only slightly stimulated by B-HT 958 or by the D1- receptor agonist SKF 38393 but it was markedly increased by the two drugs given in combination. The effect of B-HT 958 peaked earlier following low rather than high doses. The enhanced motor activity was inhibited by the D2- receptor antagonist sulpiride or the D1- receptor antagonist SCH 23390, indicating that it was caused by stimulation of both receptor types. The results suggest that B-HT 958 stimulates postsynaptic D2 receptors in addition to D2- autoreceptors and that its blockade of postsynaptic alpha 2-adrenoceptors is of no importance for the motor activity. PMID- 2570859 TI - A high dose of MPTP overcomes the protective effect of selegiline against dopaminergic neurotoxicity. AB - 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) at 90 mg kg-1 s.c., a dose lethal in non-pretreated mice, was well tolerated in selegiline [-)-deprenyl) pretreated mice and produced persistent depletion of striatal dopamine and its metabolites one week after the last of four daily injections. The protective effect of selegiline against dopaminergic neurotoxicity of MPTP can thus be overridden by a high dose of MPTP that would be lethal without selegiline pretreatment. PMID- 2570860 TI - Effect of [D-Ala2,MePhe4,Gly-ol5]enkephalin and [D-Ala2,D-Leu5]enkephalin on ion and amino acid transport in rabbit ileum. AB - The selective mu-opioid agonist [D-Ala2,MePhe4,Gly-ol5]enkephalin (DAGO) (10 microM) reduced the short circuit (Isc) and the L-valine induced increase of the transepithelial potential difference and Isc(delta Vms and delta Isc) measured in vitro in rabbit ileum, with a mechanism antagonized by naloxone (1 microM). [D Ala2,D-Leu5]enkephalin (DADLE) (10 microM) had no significant effect on the transepithelial potential difference (Vms), Isc, delta Vms and delta Isc. In the ileum deprived of the serosa and muscolaris, DAGO reduced the delta Vms and delta Isc, but not the Vms and Isc, suggesting localization of the receptors responsible for this latter effect in the myenteric plexus and/or the muscularis mucosae. These preliminary results suggest that in the rabbit ileum opioids influence electrolyte and amino acid transport and these effects may be at least partly mediated by mu-receptors. PMID- 2570861 TI - Hyperglycaemia as a factor affecting kappa-opiate agonist-induced inhibition of the gastrointestinal transit in mice. AB - The effects of highly selective kappa-opiate agonists were assessed on the gastrointestinal motility in normoglycaemic and hyperglycaemic conditions in mice. Chronic hyperglycaemia was induced by streptozocin injection (200 mg kg-1 i.p.), 7-8 days before the experiment. Acute hyperglycaemia was induced by glucose injection (5 g kg-1 i.p.) at the time of opiate administration. The kappa opiate agonists, U-50488H and U-69593 (1, 3 and 10 mg kg-1) were injected (i.p.) just before the charcoal meal. The animals were killed 45 min later and the distance travelled by the test meal was measured. In the normoglycaemic mice, both kappa-agonists significantly (P less than 0.05) inhibited the meal transit and this effect was significantly (P less than 0.05) augmented in acute hyperglycaemic animals. However, in chronic hyperglycaemic animals U-50488H failed to inhibit the charcoal meal transit, while U-69593 produced anti-transit effect comparable to that observed in normoglycaemic mice. These results demonstrate that kappa-opiate agonists produce anti-transit effects in mice that these effects are enhanced during acute hyperglycaemia. The disparity of anti transit effects of kappa-opiate agonists in acute vs chronic hyperglycaemia supports the hypothesis that elevated glucose levels are not the primary mechanism for the altered response to opiates observed in the experimental models of diabetes. PMID- 2570862 TI - Effect of reserpine pretreatment on brown fat iodothyronine 5'-deiodinase of mouse. AB - The effect of pretreatment with reserpine (1 mg kg-1 i.p. daily for 7 days) on the regulation of iodothyronine 5'-deiodinase (5'D) in mouse brown adipose tissue (BAT) has been examined. 5'D activity of BAT homogenate was assessed by the in vitro formation of 3,5,3'-triiodothyronine from thyroxine and 3,3' diiodothyronine from 3,3',5'-triiodothyronine in the presence of 20 mM dithiothreitol. Reserpine treatment decreased the stimulation of BAT 5'D induced by acute cold exposure (4 degrees C, 2 h) without a significant decrease in the basal 5'D activity, whereas stimulation of BAT 5'D elicited by noradrenaline (0.4 and 0.8 mg kg-1 s.c. 2 h previously) was not augmented after reserpine treatment. Although both noradrenaline and acute cold exposure increase BAT 5'D through alpha 1-adrenoceptors, our results show that chronic reserpine treatment prevents the effect of cold, but does not induce alpha 1-adrenoceptor supersensitivity in BAT. PMID- 2570863 TI - The application of circular dichroism (CD) to a binding study of latamoxef and beta-lactamase. PMID- 2570864 TI - Studies on interleukin-1 beta induced glycosaminoglycan release from rat femoral head cartilage in-vitro. PMID- 2570865 TI - [Is this mitral valve prolapse? A case of mitral regurgitation with early systolic murmur due to early systolic prolapse of the posterior leaflet]. AB - This paper reports the findings of phonocardiograms, echocardiogram and Doppler echocardiograms in a case of a 50-year-old man with early mitral valve prolapse with an early systolic murmur. A characteristic early systolic crescendo murmur was recorded at the apex. By amyl nitrite inhalation, the early systolic murmur was attenuated and a late systolic murmur was evoked. On the contrary, methoxamine injection increased the intensity of the early systolic murmur. Early systolic prolapse and early systolic buckling were recorded by two-dimensional and M-mode echocardiography. The phase of mitral regurgitation detected by M-mode color Dopper echocardiography coincided well in timing with the early systolic murmur and the early systolic buckling recorded on the M-mode echocardiogram. A discussion was made on the mechanism of the early systolic mitral regurgitation due to early mitral valve prolapse. PMID- 2570866 TI - Renal periarterial nerve stimulation-induced vasoconstriction at low frequencies is primarily due to release of a purinergic transmitter in the rat. AB - In the isolated rat kidney, the vasoconstrictor response elicited by periarterial nerve stimulation at low frequencies (2 Hz) is resistant to alpha adrenergic receptor blockade. It has been proposed that in some blood vessels ATP is coreleased with norepinephrine during nerve stimulation to activate P2-purinergic receptors and is responsible for the component of the vasoconstrictor response that is resistant to alpha adrenergic receptor blockade. To assess the contribution of a purinergic transmitter in the vasoconstriction elicited by periarterial nerve stimulation in the isolated Tyrodes-perfused rat kidney, fractional overflow of [3H]norepinephrine and vasoconstrictor responses to renal nerve stimulation were examined after alpha adrenergic receptor blockade and/or P2-purinergic receptor desensitization. The alpha-1 adrenergic receptor antagonists prazosin (0.1-1.0 microM) and corynanthine (0.1-1.0 microM) and the nonselective alpha adrenergic receptor antagonist phentolamine (0.1-1.0 microM) did not significantly reduce vasoconstrictor responses elicited by low frequency (0.5-4 Hz) but attenuated the responses to high-frequency (6-10 Hz) periarterial nerve stimulation. At low-frequency renal nerve stimulation, selective P2 purinergic receptor desensitization abolished the vasoconstriction at 0.5 Hz and dramatically attenuated the responses up to 4 Hz. In the presence of prazosin, the component of the vasoconstrictor response that was resistant to alpha adrenergic receptor blockade at all frequencies of renal nerve stimulation was abolished after treatment with alpha, beta-methylene ATP. On the other hand, in the isolated perfused rabbit kidney, prazosin (1.0 microM) alone reduced dramatically the vasoconstrictor responses to periarterial nerve stimulation over the same frequencies used in the rat.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570867 TI - Discriminative stimulus properties of histamine H1-antagonists in animals trained to discriminate d-amphetamine or pentobarbital. AB - Pigeons were trained to discriminate i.m. administered d-amphetamine (AMPH) or pentobarbital (PB) from saline with responding maintained under a fixed-ratio 30 schedule of food delivery. Rhesus monkeys were trained to discriminate intragastrically administered AMPH or PB from saline using a signaled shock avoidance trial procedure. In AMPH-trained pigeons the histamine H1-antagonists tripelennamine, diphenhydramine and chlorpheniramine consistently produced greater than 80% AMPH-appropriate responding. Pyrilamine substituted for AMPH in two of three pigeons. In contrast, chlorcyclizine, hydroxyzine, promethazine and the histamine H2-antagonist cimetidine all failed to produce AMPH-appropriate responding. None of the histamine H1-antagonists tested substituted for PB in PB trained pigeons. In AMPH-trained monkeys, only tripelennamine completely substituted for AMPH. Whereas chlorpheniramine, diphenhydramine and pyrilamine did not produce AMPH-appropriate responding in monkeys, these compounds did produce observable excitation and convulsions. As with the PB-trained pigeons, none of the histamine H1-antagonists tested substituted for PB in monkeys. The results of the present study demonstrate that histamine H1-antagonists have differential discriminative stimulus properties in both pigeons and monkeys. Specifically, histamine H1-antagonists known to produce more central nervous system stimulation in humans share discriminative stimulus properties with AMPH and/or produce observable signs of stimulation in monkeys.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2570868 TI - Role of mu and kappa opioid receptors in conditional fear-induced analgesia: the antagonistic actions of nor-binaltorphimine and the cyclic somatostatin octapeptide, Cys2Tyr3Orn5Pen7-amide. AB - When rats are returned to an environment in which they previously received electric shock they show a reduction in sensitivity to painful stimuli. This conditional fear-induced analgesia was measured using the formalin test. The octapeptide Cys2Tyr3Orn5Pen7-amide (CTOP; 0, 10, 40 and 160 ng/rat) dose dependently reversed conditional analgesia when administered i.c.v. The 40-ng dose partially attenuated fear-induced analgesia, whereas the 160-ng dose reversed it completely. Using similar procedures, CTOP was tested for its ability to reverse the analgesia produced by i.c.v. administered [D-Ala2,-NMPhe4, Glyol5] enkephalin, [D-Pen2,D-Pen5]-enkephalin and U50488H, which are highly selective opioid agonists at mu, delta and kappa receptors, respectively. At 40 ng/rat, CTOP reversed the analgesia produced by the mu selective ligand but not that produced by the delta ligand or the kappa ligand. At 80 ng CTOP antagonized the analgesia produced both by both enkephalin analogs but not U50488H. Nor binaltorphimine (0, 1, 3, 10 and 30 micrograms/rat) had no effect on conditional analgesia. However, the 10- and 30-micrograms doses could reverse completely the analgesia produced by U50488H. Therefore, it appears that mu and delta, but not kappa receptors, are involved in this opioid form of stress-induced analgesia. PMID- 2570869 TI - Is L-glutamate a neurotransmitter of baroreceptor information in the nucleus of the tractus solitarius? AB - The nucleus of the tractus solitarius (NTS) is the central termination site of peripheral baroreceptor afferents. It has been suggested that the excitatory amino acid (EAA) L-glutamate may be a neurotransmitter which mediates transmission of baroreceptor information in the rat NTS. The present studies examined this hypothesis by pharmacological blockade of EAA receptors in the NTS. Bilateral microinjections into the NTS of the EAA receptor antagonist kynurenic acid (KYN) virtually abolished baroreflexes elicited by electrical stimulation of the aortic nerve. In addition, hypotension and bradycardia evoked by injections into the NTS of the selective EAA receptor agonists N-methyl-D-aspartate, kainate and the quisqualate-selective agonist, alpha-amino-3-hydroxy-5-methylisoxazole-4 proprionic acid were reduced by 75% or more after KYN. Paradoxically, KYN did not reduce cardiovascular responses produced by injections into the NTS of L glutamate. These results demonstrate that 1) blockade of EAA receptors in the NTS is sufficient to abolish synaptically mediated aortic baroreflexes; 2) cardiovascular responses evoked by L-glutamate are not reduced after documented blockade of N-methyl-D-aspartate, kainate and quisqualate receptors in the NTS by KYN; and 3) under the conditions of these experiments, the pharmacological actions of exogenously administered L-glutamate are not identical to those of the neurotransmitter which is released at NTS synapses by aortic nerve stimulation. These observations suggest that an EAA or EAA analog other than L-glutamate may be the neurotransmitter of baroreceptor information in the NTS. PMID- 2570870 TI - Are changes in neuronal calcium channels involved in ethanol tolerance? AB - Effects on the development of tolerance to ethanol of concurrent administration of the dihydropyridine calcium channel antagonist, nitrendipine, were investigated. Ethanol tolerance was induced in rats by repeated i.p. injections and measured by performance on the rotarod apparatus. Concurrent injections of nitrendipine prevented the development of tolerance to the ataxic action of ethanol. A similar effect was seen with the calcium channel antagonist PN 200-110 on tolerance to the general anesthetic effects of ethanol in mice. As previously reported, chronic ethanol administration increased the number of dihydropyridine binding sites in rat cerebral cortex. This was also prevented by the nitrendipine treatment; repeated administration of both ethanol and nitrendipine caused a decrease in the number of binding sites. The effect of nitrendipine on the tolerance was not due to alterations in body temperature, changes in blood pressure, differences in brain ethanol concentrations or residual nitrendipine in the brain at the time of testing. The results suggest that alterations in dihydropyridine-sensitive calcium channels may be involved in the adaptations that occur on chronic treatment with ethanol. PMID- 2570872 TI - Rheumatoid arthritis and AIDS. PMID- 2570873 TI - Beta-blockers with ACE inhibitors--a logical combination? AB - Although there is no obvious pharmacological rationale for the combined use of an ACE inhibitor and a beta-blocker in the treatment of hypertension, this combination has proved to be more effective than monotherapy in a number of studies, some of which are reviewed. Moreover, there is evidence that this combination may be beneficial after myocardial infarction. Combined use of ACE inhibitor and beta-blocker therapy therefore warrants further investigation. PMID- 2570871 TI - Electrophysiological responses to somatostatin of rat hypophysial cells in somatotroph-enriched primary cultures. AB - 1. Somatotroph cells were obtained from pituitaries of adult male rats by dissociation, separation and enrichment on a continuous gradient of bovine serum albumin at unit gravity. They were kept in culture for 7-15 days before electrophysiological experiments. 2. Immunofluorescent staining of the resulting gradient fractions (numbered F2 to F9) indicated that the majority of somatotrophs (75-85%) were located in the heavy fractions (F8 and F9). However, a small percentage (15-20%) of cells in these fractions were identified as lactotrophs. 3. Perifusion experiments indicated that on the one hand release of growth hormone from somatotroph-enriched fractions was stable at the level of 6 ng (2 min)-1 (10(6) cells)-1 and was markedly inhibited by somatostatin (1.9 ng (2 min)-1 (10(6) cells)-1) but not by dopamine. On the other hand, in the same cell preparations, basal prolactin release (1.6 ng (2 min)-1 (10(6) cells)-1) was significantly reduced by dopamine (0.08 ng (2 min)-1 (10(6) cells)-1) but remained unchanged by somatostatin treatment. 4. The inhibitory effect of somatostatin on growth hormone release was dose dependent. This effect was not abolished by tetraethylammonium (40 mM) or 4-aminopyridine (5 mM), but somatostatin decreased high-potassium-induced release. 5. In all the cells recorded (n = 187), 14% (n = 26) displayed a low resting potential (less than -30 mV) and poor membrane resistance (less than 50 M omega). The recording was unstable and resting potentials decreased regularly to 0 mV in less than 5 min. The other 86% of the cells displayed resting potentials varying from -45 to -65 mV and had a membrane resistance of more than 150 M omega. Only cells which displayed these membrane characteristics showed clear responses to somatostatin or dopamine, and were therefore chosen for experiments. 6. In all the cells selected for the experiments (n = 161), 78% (n = 126) showed either triggered or spontaneous action potentials. The action potentials remained insensitive to sodium-free bath solution, but were reversibly blocked by the calcium channel blockers cobalt (5 mM) or nickel (5 mM). 7. When the cells were at resting potential, somatostatin induced a hyperpolarizing response associated with a decrease of membrane resistance. During this response, spontaneous or triggered action potentials were inhibited. The hyperpolarizing response induced by somatostatin was dose-dependent.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2570874 TI - Characterisation of a fimbrial, mannose-resistant and eluting haemagglutinin (MREHA) produced by strains of Salmonella of serotype Sendai. AB - Strains of Salmonella of serotype Sendai, producing a mannose-resistant and eluting haemagglutinin (MREHA) when cultured at 37 degrees C but not at 18 degrees C, were examined by electronmicroscopy after negative staining. Production of this MREHA, previously thought to be nonfimbrial, was correlated with the presence of thick fimbriae with an external diameter of 13.6 nm. These fimbriae were readily fragmented and, when purified, had an estimated Mr of 28 Kda. Production of fimbrial MREHA by Sendai strains was associated with the ability to adhere to a wide range of substrates and to form a fimbrial pellicle at the surface of liquid media incubated statically in air. The origin of this unusual Sendai fimbrial MREHA is unknown. Thin filamentous structures produced independently of fimbrial MREHA by Sendai strains were also described. Fimbrial MREHA was not produced by strains of the antigenically similar serotype Miami which, however, and unlike Sendai strains, formed mannose-sensitive haemagglutinin and type-1 fimbriae. The ability to differentiate strains of Miami and Sendai (serotype 1,9,12:a: 1,5) by means of their fimbriae is noted. PMID- 2570875 TI - New views of human cardiac beta-adrenoceptors. PMID- 2570876 TI - Positive inotropic effect of somatostatin in guinea-pig ventricular muscles. AB - We investigated the effect of somatostatin (SS) in guinea-pig ventricular muscles using a force transducer and a conventional microelectrode method. Instead of a negative inotropic effect in atrial muscles, SS (10(-11) to 10(-7) M) elicited a positive inotropic effect in ventricular muscles in a concentration-dependent fashion, without changing the time course of contraction. The positive inotropic effect of SS was accompanied by a significant enhancement of the slow action potentials and was suppressed by diltiazem and phentolamine. An increase of extracellular Ca2+ concentration or stimulation frequency enhanced the positive inotropic effect of SS. The positive inotropic effect of SS was not suppressed in the presence of propranolol, metoclopramide, cimetidine or indomethacin, and it appeared even under cold conditions. These results suggest that SS has a positive inotropic effect in guinea-pig ventricular muscle, which is at least partly due to an increase in the slow inward Ca2+ current. PMID- 2570877 TI - Involutional paranoid disorder: a forgotten syndrome. AB - Involutional paranoid disorder is seen frequently in medical and psychiatric inpatient units, geriatric units, and other medical settings in which persons in the climacteric age group are treated. Little information is provided in the recent literature to guide the practitioner in diagnosing and treating this disorder. The authors review the clinical features of involutional paranoia and present three case reports to illustrate the characteristic symptoms. Diagnostic and treatment considerations are also reviewed. PMID- 2570878 TI - Lack of modifying effects of 2,2'-[(4-aminophenyl) imino] bisethanol sulfate on induction of preneoplastic gamma-glutamyl transpeptidase-positive foci in a medium-term bioassay system using F344 rats. AB - The modifying effects of 2,2'-[(4-aminophenyl) imino] bisethanol sulfate (4APE) on liver carcinogenesis were investigated in male F344/DuCrj rats initially treated with N-nitrosodiethylamine (DEN). Two weeks after a single dose of DEN (200 mg/kg, intraperitoneally), rats were given 4APE at dietary levels of 1,000, 330 and 110 parts per million (ppm), or 3'-methyl-4-dimethylaminoazobenzene (3' Me-DAB) at 600 ppm as a positive control for 6 weeks. At week 3 following DEN administration, all animals were subjected to partial hepatectomy. No adverse effects on survival and body weight were seen in rats treated with 4APE, even at the highest dietary levels. Marked growth retardation and significant increase of relative liver weight were found in animals treated with the known hepatocarcinogen 3'-Me-DAB, which demonstrated marked promoting activity as evidenced by significantly increased values for gamma-glutamyl transpeptidase (gamma-GT) positive foci as compared with the controls given DEN alone. In contrast, 4APE did not significantly increase the level of gamma-GT positive foci over that induced by DEN initiation alone. From these results, it is concluded that 4APE does not possess promoting activity for liver carcinogenesis. PMID- 2570879 TI - Opacification test by using the pig isolated cornea and its application to a test of corneal opacity induced by befunolol hydrochloride. AB - Corneas isolated from the pig eyeball have been treated in vitro with varying concentrations of befunolol hydrochloride, in order to test the topical effect of befunolol on the cornea in terms of opacity. Experiments were carried out with the corneas treated as follows: intact, epithelium-removed, endothelium-removed, and both epithelium- and endothelium-removed. Solutions of the drug were applied to both epithelial and endothelial surfaces, to the epithelial surface only, or to the endothelial surface only. When the drug was applied either to both surfaces or to the endothelial surface only, there was a significant increase in opacity. However, when applied to the epithelial surface only, befunolol caused an insignificant increase in opacity as compared with that of control. It is suggested, therefore, that corneal opacity due to befunolol hydrochloride topically administered to the eye, is concerned mainly with the epithelium in cornea, since the drug is uncapable of passing barrier probably existing in surface layer of epithelium but it easily penetrates in corneal layers from endothelial side to attain reversely to epithelial cells. Thus, it is probable that the risk to induce opacity is minor in case that befunolol is topically given in drops to the eye. PMID- 2570880 TI - Effects of afloqualone on vestibular nystagmus and the lateral vestibular nucleus. AB - To clarify the antivertiginous effect of afloqualone, an antispastic drug, we examined its action on the vestibular nervous system in cats. The results suggest that afloqualone inhibits vestibular nystagmus probably due to both inhibition of selective polysynaptic transmission and enhancement of the effects of GABA and glycine in the lateral vestibular nucleus (LVN), and its GABA-enhancing effect is thought to be attributable to the increased sensitivity of GABA receptors of the LVN neuron site. PMID- 2570881 TI - [Atherogenic properties of beta adrenergic blockaders evident on vascular wall cells]. AB - In cultured human cells, beta-adrenoblockers such as propranolol, alprenolol, metoprolol, atenolol, pindolol, and thymolol, as well as sera from patients with coronary heart disease were examined for atherogenic activity following a single administration of propranolol (80 mg) and pindolol (10 mg). Addition of the beta adrenoblockers to the cell culture was demonstrated to enhance cellular total cholesterol levels and to stimulation their proliferation. Propranolol and pindolol given in a single dose was found to result in the appearance of atherogenic properties of the patients' sera. PMID- 2570882 TI - [Autologous peripheral stem cell transplantation in children]. AB - 19 children between 3 and 23 years underwent 79 leukapheres for collection of blood stem cells. In children suffering from acute lymphoblastic leukemia (ALL), Non Hodgkin's Lymphoma (NHL) and Ewing's Sarcoma (ES) we collected 6.87 x 10(4) CFU-GM/kg (range 2,65-21.7), if collections were started with the first platelet rise. In children with peripheral primitive neuroectodermal tumors (PNET) and neuroblastoma (NBL) we gained only 1.20 x 10(4) CFU-GM/kg (range 0.09-2.24). 17 children received high dose chemoradiotherapy and peripheral stem cell +/- bone marrow rescue. 9 suffered from solid tumors, 8 from hematopoietic malignancies. 9 were transfused with peripheral stem cells only, 8 received bone marrow in addition. Time to reach 0.5 x 10(9)/l granulocytes was very short-median 31 days (12-65), in 4 children receiving more than 5 x 10(4) CFU-GM/kg 12 to 13 days, only. On January 31st, 1989 6/17 children are alive in complete remission after a median observation time of 14.5 months (3-26) after autologous stem cell transfusion, one child is alive in "no remission", 7 died with relapse, 3 died because of infections (2 x aspergillosis, 1 x pseudomonas septicemia). The collection of blood derived stem cells by leukaphereses was well tolerated even in very small children and easily repeatable. With optimal timing high stem cell numbers were obtainable, resulting in a very short duration of posttransplant granulocytopenia. PMID- 2570883 TI - [Use of strong opioids in the treatment of cancer pain in adults and children]. AB - A high percentage of adults and children with advanced cancer suffer from pain. Strong opioids for pain control, e.g. morphine, slow-released morphine or buprenorphine, should be administered early according to the intensity of pain. The analgesics should be given orally whenever possible. They must be given at fixed intervals based on the duration of their action. The dose must be titrated to the needs of the patient. Sometimes more than 1000 mg morphine orally per day is necessary. Correctly used strong opioids produce only a few side-effects, especially constipation and vomiting. Many studies in adult cancer patients all over the world demonstrate the effectiveness of strong opioids for pain control. Children should be treated in the same way and comparable data in children with cancer pain must be collected. PMID- 2570884 TI - Neurotransmitter release: development of a theory for total release based on kinetics. AB - According to the calcium-voltage hypothesis for the control of neurotransmitter release, a molecule (or molecular complex) must be activated by membrane depolarization, after which the activated molecule can bind calcium and initiate release. In this study, we have examined properties of the kinetics of phasic release resulting from a set of differential equations that characterize the calcium-voltage hypothesis. It was found that, in accord with experiments, an important feature is the approximate constancy of the shape of the graph for the kinetics of phasic release at various depolarizations and extracellular calcium concentrations. The shape constancy allowed us to obtain an explicit and relatively simple analytical formula for the total transmitter release (quantal content) by approximating the differential equations of the model. This formula shows a saturating sigmoidal dependence on both intracellular and extracellular calcium concentrations. The formula thus agrees with various experiments. Moreover, it agrees with, and provides meaning to, earlier phenomenological expressions for the dependence of release on calcium concentration. In particular, the formula provides an expression for the maximal release in terms of kinetic parameters from the calcium-voltage model, and thereby allows one to supplement earlier kinetic tests of the calcium-voltage hypothesis with further tests focused upon the dependence of total release on depolarization. PMID- 2570885 TI - [A new treatment possibility for osteoporosis with osteochin (ipriflavone) tablets]. AB - It was found as a result of preclinical experimental examinations, performed in Hungary and Japan that Ipriflavone stimulates calcitonin secretion through increasing the oestrogene effect. Its effect of hindering bone resorption works presumably also this way. Authors report on experiences and results gained in 408 treatment years of 114 primary and secondary osteoporotic patients. The results of clinical experiments in Hungary, Japan and Italy are similar. The treatment with Osteochin and calcium is an effective therapy of osteoporosis. The progression of the loss of bone mass can be hindered, the number of bone fracture is diminished in proportion of time and the method proved to be effective in 79 per cent of the patients. Out of the gastric complaints, found in 5.5 per cent of the cases no side effect was observed. PMID- 2570886 TI - [Role of orthopedic surgery in the management of rheumatoid arthritis]. AB - After describing a few pathological and clinical characteristics of the rheumatoid arthritis author outlines some of the cares connected with the operative treatment of these, in the majority severe cases. The more frequently used operations and their results to be expected are described. PMID- 2570887 TI - [Experience with total hip prosthesis]. AB - Authors report on their material of total hip endoprosthesis, performed 30 May 1969--31 August 1988 on the Department of Orthopaedics of the Semmelweis University of Medical Science. In this period 1480 operations were made. The formerly and recently used operative technique and the types of the prostheses are described. The intraoperative, the early and the late complications and their treatment are also described. The formerly and recently used tactics of reoperation is reported and attention is called to the fact that the success rate of the replantation can be increased by an adequate technical skill. PMID- 2570888 TI - [Experience with the cementless hip prosthesis, with special reference to the implantation of prosthesis stems]. AB - Authors report on their experiences gained during 7 years with 102 PM (Aesculap) and 76 Etropal cementless hip endoprosthesis and with the implantation without sticking of 264 standard prosthesis stems. They explain their results with bone rebuilding that can be certified also by Roentgenograms. The lack of success in a few per cent of the cases can be explained mainly by technical insufficience. PMID- 2570889 TI - [Surgical management of lumbar spondylolisthesis]. AB - The basic principles of the operative treatment of lumbar spondylolisthesis became uniform in the literature in the last years. Authors deal shortly with some questions of the indication and of the preoperative examination and describe shortly the basic principles of the operative treatment (decompression, reduction, stabilization). The operative methods used by them are reported and illustrated (Morscher's "hookscrew" method, fusion according to Harrington-Vidal, Cloward's posterior corpodesis--PLIF--and Steffee's transpedicular system). PMID- 2570890 TI - [Experience with giant cell tumors]. AB - Authors describe, 64 cases of giant-cell tumour. 5 (9 per cent) was found in the latent, 32 (50 per cent) in the active and 27 (41 per cent) in the aggressive stage when first seen. The operative methods are described and these are determined first of all by the surgical stage of the giant-cell tumour, its size and localization, by the age of the patient and whether primary tumour or a recurrence is found. Based on their experiences, excochleation and filing is suggested in the latent and active stage of this tumour. In tumours in the agressive stage the so called wide resection in the healthy tissue is reasonable. In the rare malignant giant-cell tumour radical resection is suggested. Recurrence was found in 29 per cent after the first operation. Follow-up examinations show that 64 per cent of the patients is symptom free, in 33 per cent the main source of complaints is the consecutive arthrosis, 2 patients were lost in consequence of lung metastasis. PMID- 2570891 TI - [Surgical reposition in the management of epiphysiolysis of the femoral head]. AB - In major slipped femoral capital epiphysis the hip function can be restored with a possibly adequate correction of the deformity. Previously authors have performed various inter- and subtrochanteric femoral osteotomies with this aim, in 4 recent cases however the slipped femoral head was reduced operatively according to Dunn and Angel. 2-4 years after the operation good results were found. The blood supply of the epiphysis was not damaged in either case, and the hip motion was restricted in one case only, in which previously 2 other operations were already performed. The operative reduction with the technique described is suggested where the conditions of this considerable operation are given. PMID- 2570892 TI - [Rare manifestations of Perthes disease]. AB - Authors briefly summarize the rare manifestations of Perthes' disease. The bilateral appearance, the possibility of its connection with constitutional disorders, the manifestation on hips already damaged, the forms appearing earlier or later than usual are described. The prognostical and the therapeutic significance of recognizing these forms of Perthes disease is stressed. PMID- 2570893 TI - [Perthes disease and relief of weight bearing]. AB - Authors compare in two different groups of treatment the results of varus derotation osteotomies in Perthes' patients treated on the Department of Orthopaedics of the Pecs University of Medicine in 1974-1984. In 30 cases running was allowed after the osteotomy whereas in 52 cases an apparatus to relieve of weight bearing was used. It is stated after the comparison of results, considering also the role of the "risk" factors, influencing the results that the relieve of weight bearing does not influence the end result. PMID- 2570894 TI - [Treatment and rehabilitation of dysmelic children]. AB - It is reasonable to make the plan of treatment and rehabilitation of dysmelic children in infancy and to realize it after an agreement with the parents, if possible in a care system. A careful operative indication considering the possibilities of orthetic-prothetic supply from every respect is necessary. The supply of the orthopaedic apparatus is generally individually planned and it succeeds only rarely to use standard prosthesis and orthesis. It is worthwhile to strive that the rehabilitation procedure should enable the child to go to the general school in the usual time. Based on the presentation of a few cases the author describes the ground principles of treatment-rehabilitation summarized in the paper. PMID- 2570895 TI - bcr rearrangement: potential false positive secondary to an Eco RI restriction fragment length polymorphism. AB - Molecular studies have demonstrated that the Philadelphia chromosome (Ph) translocation characteristic of chronic granulocytic leukemia (CGL) and 50% of the cases of Ph positive acute lymphocytic leukemia (ALL) involves a limited 5.8 Kb region on chromosome 22 termed the breakpoint cluster region (bcr). Detection of bcr rearrangement by Southern blot analysis has proven to be a sensitive diagnostic method and can identify this translocation in some cases which appear cytogenetically negative. Restriction fragment length polymorphisms (RFLP) which involve bcr have the potential to be misinterpreted as gene rearrangements since they result in alteration of the DNA fragment size detected by Southern blot hybridization. We have identified a RFLP involving bcr that is detectable with Eco RI digestion but not with Bam HI, BgI II, or Xba I. The polymorphic fragments generated indicate that this RFLP is the result of an Eco RI restriction site sequence polymorphism. PMID- 2570896 TI - [Recommendations from a meeting of experts: Neuroleptics--rational use in the treatment of psychoses and geriatrics]. PMID- 2570897 TI - [The 5th International Conference on AIDS. The HIV cell receptor is more and more interesting in the research on drugs against AIDS]. PMID- 2570898 TI - Duration of human immunodeficiency virus infection before detection of antibody. AB - To estimate the duration and frequency of the period of HIV infection without detectable antibody, modelling techniques were applied to results of detection of HIV DNA by means of the polymerase chain reaction (PCR) and to data from cases in published reports. PCR was carried out with gag and env region primers on samples from 27 homosexual and 12 haemophilic men for whom stored samples were available from before and after seroconversion; serum was also tested for p24 antigen by antigen-capture enzyme immunoassay. HIV DNA was detectable before seroconversion in 4 men; in all 4 PCR was positive only in the seronegative sample taken closest to the time of seroconversion. In 3 men antigen was detected before seroconversion; in each case HIV DNA was also detected. By a Markov model, the time from infection with HIV (as assessed by detection of HIV DNA) to first detection of HIV antibody was estimated to be 2.4 (SE 2.1) months for the median individual. Modelling of cases of HIV infection with known exposure in published reports gave a median estimate of 2.1 (0.1) months from exposure to antibody detection, and 95% of cases would be expected to seroconvert within 5.8 (0.6) months. HIV infection for longer than 6 months without detectable antibody seems uncommon. PMID- 2570899 TI - Specificity of calcium channel autoantibodies in Lambert-Eaton myasthenic syndrome. AB - Autoantibodies that interfere with neurotransmitter release by affecting the function of voltage-operated calcium channels (VOCCs) have been found in patients with Lambert-Eaton myasthenic syndrome (LES). To find out the nature of the antigen to which these autoantibodies bind, tests were done with omega-conotoxin, which blocks some types of VOCCs. LES antibodies were able to immunoprecipitate VOCCs prepared from the human neuronal cell line IMR32 which were pre-labelled with the specific VOCC ligand omega-conotoxin. LES autoantibodies are also able to specifically down-regulate the expression of VOCCs in IMR32 cells. A new radioimmunoassay for the quantitative detection of LES antibodies was developed and found to be of value in distinguishing LES patients from patients with myasthenia gravis and some other neurological disorders. PMID- 2570900 TI - Smoking as a risk factor for cerebral ischaemia. AB - To assess whether a rigorous clinical classification, based on computerised tomography, of patients with cerebral ischaemia would identify subgroups at higher or lower risk with respect to cigarette smoking habits, a case-control study was carried out on 422 cases of first-episode cerebral ischaemia matched for age and sex with 422 community-based neighbourhood controls. Patients with ischaemic stroke due to extracranial or intracranial vascular disease were at higher risk from smoking than has previously been reported for stroke (relative risk 5.7, 95% confidence interval 2.8, 12.0) whereas those with stroke due to cardiac emboli had no excess risk associated with smoking (relative risk 0.4 [0.1, 1.8]). After cessation of smoking, the relative risk declined gradually over 10 years, at the end of which time a significant risk was still evident. This finding may imply that the risk incurred by smoking is due mainly to atheroma formation, rather than transient haematological effects. Exposure to smoking by a spouse was an independent risk factor for the whole group of cerebral ischaemia patients (relative risk 1.7 [1.1, 2.6]), but this was not so for smoking by either parent (relative risk 1.2 [0.8, 1.8]). These findings suggest that smoking is a more potent risk factor for the most common form of ischaemic stroke than has previously been appreciated. The persistent nature of the risk even after cessation of smoking and the possible risk associated with passive exposure strengthens public health arguments against smoking. PMID- 2570901 TI - Percutaneous coronary excimer laser angioplasty: initial clinical results. AB - A novel 1.3 mm diameter laser catheter, consisting of 20 concentric 100 microns quartz fibres around a central lumen for a 0.35 mm flexible guide wire, was used to ablate atherosclerotic tissue in thirty patients with coronary artery disease. The laser catheter was coupled to an excimer laser delivering energy at a wavelength of 308 nm and a pulsewidth of 60 ns. The primary success rate was 90% (27 of 30 lesions). The mean (SD) percentage stenosis fell from 85 (15)% to 41 (19)% after laser ablation. In ten patients the lumen diameter after laser angioplasty was considered sufficient, but subsequent balloon angioplasty was carried out for the other twenty patients. Failure to pass the lesion was caused by vessel kinking in two patients and a total occlusion in one patient. No complications directly attributable to laser ablation, such as vessel wall perforation, occurred; one dissection occurred but had no clinical sequelae. There was one early reocclusion and death in a patient with triple vessel disease and unstable angina, probably as a result of plaque rupture after balloon angioplasty. These results are encouraging and justify further clinical investigations. PMID- 2570902 TI - In-vivo radiotoxicity of DNA-incorporated 125I compared with that of densely ionising alpha-particles. AB - When the atomic nucleus of 125I decays by orbital electron capture followed by internal conversion, numerous very-low-energy electrons (Auger electrons) are emitted, so that the energy density in the immediate vicinity of the decay site is extremely high. 125I incorporated into DNA was as effective as densely ionising 5.3 MeV alpha-particles from 210Po in reducing the sperm-head population in mice. Hence the biological risks of Auger-electron emitting radionuclides widely used in biology and medicine ought to be reassessed. PMID- 2570903 TI - Reperfusion injury after thrombolytic therapy for acute myocardial infarction. PMID- 2570904 TI - Obstetrics and social change. PMID- 2570905 TI - Xanthochromia. PMID- 2570907 TI - Changes in ventilation during V/Q scanning. PMID- 2570906 TI - Swallowed coins. PMID- 2570908 TI - Topical iodine-containing antiseptics and neonatal hypothyroidism in very-low birthweight infants. AB - The thyroid function of very-low-birthweight (VLBW; below 1500 g) infants admitted to neonatal intensive-care units was studied at two hospitals; one routinely used topical iodinated antiseptic agents and the other used chlorhexidine-containing antiseptics. Serial monitoring of urinary iodine excretion and serum thyrotropin and thyroxine levels was undertaken from birth for the first 4 weeks of life. Urinary iodine excretion rose dramatically in the 54 iodine-exposed infants and was up to fifty times greater than in the 29 non exposed infants. Within 14 days, 25% (9 of 36) of the infants exposed to iodine had serum thyrotropin levels above 20 mIU/l, compared with none of the control group. The mean serum thyroxine level in these 9 infants (44.1 nmol/l) was significantly lower than that in exposed infants with normal thyrotropin levels (83.1 nmol/l) and in the non-exposed control group (83.0 nmol/l); thyroxine levels fell before serum thyrotropin rose. These disturbances in thyroid function correlated positively with urinary iodine excretion and hence iodine absorption. Thyroid function had returned to normal by the time of discharge from hospital. It is concluded that iodine absorption, from topical iodine-containing antiseptics, may cause hypothyroidism during a critical period of neurological development in the newborn infant. The routine use of iodine antisepsis in VLBW infants should be avoided because of this effect. PMID- 2570909 TI - Radiotherapy for Paget's disease of the nipple: a conservative alternative. AB - The case records of 28 patients with Paget's disease of the nipple treated by radio-therapy alone were reviewed retrospectively. 16 of 19 patients who had no palpable underlying tumour and who were mammographically normal at the time of original treatment remain free of disease with a median follow-up of 5 years 3 months. In this selected group, radical radiotherapy with small fields localised to the involved skin is an effective alternative to mastectomy. PMID- 2570910 TI - Haematological indices in Nigerians exposed to radioactive waste. AB - The haematological indices in 26 workers who came in contact with radioactive and toxic wastes dumped in Koko village, Nigeria, were compared with those in 26 workers at a port 1 km away from the dump site and with those in 26 controls derived from Port Harcourt, which is about 200 km from the dump site. The site workers showed a macrocytic anaemia and a leucopenia that affected the minority components rather than the neutrophils or lymphocytes. Radiation injury was the likely cause of the findings in the site workers. PMID- 2570911 TI - The continuing hazard of lead in drinking water. PMID- 2570913 TI - Duct carcinoma-in-situ: can we predict recurrence after surgery? PMID- 2570912 TI - Localisation of focal lesion permitting partial pancreatectomy in infants. PMID- 2570914 TI - Gastrospirillum hominis infection of the stomach. PMID- 2570915 TI - Occult blood tests. PMID- 2570916 TI - Early marker for Alzheimer's disease: the atrophic hippocampus. PMID- 2570917 TI - Radon and leukaemia. PMID- 2570918 TI - Encephalopathy in patient taking aluminium-containing agents, including sucralfate. PMID- 2570919 TI - Cerebral perfusion deficits in dysbaric illness. PMID- 2570920 TI - Desmopressin and thrombosis. PMID- 2570921 TI - Left-ventricular function as an end-point. PMID- 2570922 TI - Myocardial infarction risk, earlobe crease, and sleep apnoea syndrome. PMID- 2570923 TI - Comparison of gram stain and latex agglutination for diagnosis of meningococcal meningitis. PMID- 2570924 TI - Mumps meningitis after measles, mumps, and rubella vaccination. PMID- 2570925 TI - Drug economics in developing countries. PMID- 2570926 TI - Regulating drug advertisements. PMID- 2570927 TI - Switzerland's drug surveillance. PMID- 2570928 TI - World psychiatry and the Soviet Union. PMID- 2570929 TI - Mature entrants to medicine. PMID- 2570930 TI - Treatment of suxamethonium anaphylaxis. PMID- 2570931 TI - Hazards of nasogastric tube insertion in the newborn infant. PMID- 2570932 TI - Triazolam. PMID- 2570933 TI - Cytomegalovirus infection and progression to AIDS. PMID- 2570934 TI - Iron deposition in liver in zidovudine-related transfusion-dependent anaemia. PMID- 2570935 TI - Reduced sweating in adults with growth hormone deficiency. PMID- 2570936 TI - Does buspirone have anxiogenic properties? PMID- 2570937 TI - Predominance of vancomycin-sensitive strains of Neisseria gonorrhoeae in Djibouti. PMID- 2570938 TI - Naloxone and opiate addiction. PMID- 2570939 TI - Q-fever endocarditis twenty-five years on. PMID- 2570940 TI - ELISA for anti-giardia IgM. PMID- 2570941 TI - Antibiotic treatment in bacterial meningitis. PMID- 2570942 TI - Criteria for immune trials in type I diabetes. PMID- 2570943 TI - Sex difference in D7S8 marker allele distribution in adult cystic fibrosis patients. PMID- 2570944 TI - Microencapsulated xanthine oxidase as experimental therapy in Lesch-Nyhan disease. PMID- 2570945 TI - Measles and autoimmune chronic active hepatitis. PMID- 2570946 TI - In-vitro fertilisation today. PMID- 2570947 TI - Altered sex ratios. PMID- 2570948 TI - Microsurgery for cold intolerance after finger replantation. PMID- 2570949 TI - Retinopathy after irradiation for Graves' ophthalmopathy. PMID- 2570950 TI - Ophthalmopathy and thyroid stimulation. PMID- 2570951 TI - Diarrhoea and malnutrition. PMID- 2570952 TI - Continuous intrapartum monitoring of fetal oxygen saturation. PMID- 2570953 TI - Fish oil for preventing coronary restenosis. PMID- 2570955 TI - 100,000 AIDS cases. PMID- 2570954 TI - Steroid-induced psychosis. PMID- 2570956 TI - Halving of mortality of severe melioidosis by ceftazidime. AB - An open randomised trial was conducted to compare ceftazidime (120 mg/kg/day) with "conventional therapy" (chloramphenicol 100 mg/kg/day, doxycycline 4 mg/kg/day, trimethoprim 10 mg/kg/day, and sulphamethoxazole 50 mg/kg/day) in the treatment of severe melioidosis. A paired restricted sequential trial designed to detect a reduction in mortality from 80 to 40% in culture-positive patients surviving greater than 48 hours was stopped after 22 months. Of the 161 patients entered into the study, 65 had bacteriologically confirmed melioidosis and 54 of these were septicaemic. Ceftazidime treatment was associated with a 50% (95% CI 19-81%) lower overall mortality than conventional treatment (74% vs 37%; p = 0.009) and should now become the treatment of choice for severe melioidosis. PMID- 2570957 TI - Treatment of enteric hyperoxaluria with calcium-containing organic marine hydrocolloid. AB - An organic marine hydrocolloid (OMH) charged with calcium ('Ox-Absorb') was studied in vitro for oxalate binding and in patients with enteric hyperoxaluria to investigate oxalate excretion and the inhibitory activity on crystal formation of the urine. In-vitro experiments showed complete binding of oxalate to OMH. In clinical studies in nineteen patients with intestinal disorders and stone formation, urinary oxalate excretion was significantly lower during OMH treatment than off treatment. The activity product index of calcium oxalate was reduced on treatment. A pronounced rise in the inhibitory activity of urine was seen in two patients with very low pretreatment values. Most patients experienced virtual normalisation of bowel function, and in those with severe stone formation there was substantial clinical improvement. It is concluded that OMH has the capacity to bind oxalate in vitro and to reduce urinary oxalate excretion. These observations suggest a new promising treatment for enteric hyperoxaluria. PMID- 2570958 TI - Comparison of propofol and midazolam for sedation in critically ill patients. AB - 101 critically ill patients admitted to five intensive-care units were allocated randomly to receive a continuous intravenous infusion of either propofol or midazolam for sedation for up to 24 h. In addition, morphine was given to provide analgesia. The mean duration of infusion was 20.2 h (range 3.0-24.5) in the propofol group and 21.3 h (4.0-47.0) in the midazolam group and infusion rates were 1.77 mg/kg/h (range 0.40-5.00) and 0.10 mg/kg/h (0.01-0.26), respectively. The infusion rates were adjusted as necessary, and the desired level of sedation was achieved easily in most patients in both groups. There were slight falls in arterial pressure, but there were no significant differences between the groups. Heart rate was lower in patients who received propofol. Some small changes occurred in biochemical and haematological variables in both groups, but they were not clinically significant. There was no indication that either drug substantially impaired adrenal steroidogenesis. When the infusion was discontinued, there was less variability in recovery of consciousness in patients who had received propofol. In a subgroup of patients, weaning from mechanical ventilation was achieved significantly faster after discontinuation of propofol than of midazolam. Propofol proved to be a satisfactory agent for sedation of these critically ill patients and compared favourably with midazolam. PMID- 2570959 TI - Passive immunisation of children with bovine colostrum containing antibodies to human rotavirus. AB - The efficacy of a 10-day course of bovine colostrum with high antibody titre against the four known human rotavirus serotypes in protecting children against rotavirus infection was examined in patients admitted to hospital. Children aged 3 to 15 months were blocked in pairs according to ward accommodation (ie, isolation or open area). Each block contained 1 treated and 1 control child. The allocation to treatment or control (an artificial infant formula) was randomised. 9 of 65 control children but none of 55 treated children acquired rotavirus infection during the treatment period (p less than 0.001). The importance of protecting against rotavirus infection was highlighted by the fact that parents of symptomatic rotavirus-positive children sought medical attention seven times more often than did parents of symptomatic rotavirus-negative children (p less than 0.05). PMID- 2570960 TI - Tumour necrosis factor in bronchopulmonary secretions of patients with adult respiratory distress syndrome. AB - Tumour necrosis factor (TNF) concentrations were measured in the bronchopulmonary secretions of 5 patients with the adult respiratory distress syndrome. Each patient underwent fibreoptic bronchoscopy and bronchopulmonary aspiration, and control samples were obtained in an identical manner from 24 patients who underwent bronchoscopy for other reasons (8 had tuberculosis, 6 had sarcoidosis, and 10 had haemoptysis but no abnormal findings). Aspirated fluid was assayed for the presence of TNF by use of an enzyme-linked immunosorbent assay. In the 5 patients with adult respiratory distress syndrome, TNF concentrations exceeded 500 U/ml (12.5 ng/ml), whereas in the control samples no TNF was detected. PMID- 2570961 TI - Use of digital arterial pressure to detect aortic compression during labour. PMID- 2570963 TI - Dyslexia. PMID- 2570962 TI - Drugs acting on 5-hydroxytryptamine receptors. PMID- 2570964 TI - Vascular malformations in the brainstem. PMID- 2570965 TI - Which orthosis? PMID- 2570966 TI - Autonomic neuropathy in liver disease. PMID- 2570967 TI - UK medical research handbagged again. PMID- 2570968 TI - Effect of smoking on meningococcal carriage. AB - A case-control study was done to examine whether certain environmental or medical factors were associated with meningococcal carriage. Questionnaires were posted to 138 meningococcal carriers and their controls, and to 52 carriers of Neisseria lactamica and their controls. Carriers were matched to controls by age, sex, and area of residence. The overall response rate was 89%. There were no differences in environmental or medical factors between N lactamica carriers and their controls, nor in household crowding, housing conditions, frequency of physical exercise, or upper respiratory disorders between meningococcal carriers and their controls. Active smoking and the presence of other smokers in the household were independently associated with meningococcal carriage; the risk of carriage increased significantly with heavier smoking. PMID- 2570969 TI - Association between alcohol consumption and cancer of the sigmoid colon: observations from a Japanese cohort study. AB - A 17-year follow up of 265,118 Japanese adults aged 40 years and above revealed a close association between cancer of the sigmoid colon (n = 91) and alcohol consumption: relative risk (RR) for drinkers versus non drinkers was 4.38 (90% CI 1.75-10.97) in men and 1.92 (1.13-3.26) in women. In men, attributable risk was 74% and RRs in non, infrequent, occasional, and daily drinkers were 1.00, 2.03, 3.83, and 5.42, respectively. RRs of daily consumption versus non-consumption in men for cancers of mouth, pharynx, oesophagus, stomach, proximal colon, sigmoid colon, and rectum were 2.27, 2.44, 2.29, 0.92, 0.98, 5.42, and 1.39, respectively. A combined effect of alcohol and smoking was evident for cancers of the upper digestive tract but almost absent for those of the lower digestive tract. PMID- 2570970 TI - Oedema protein concentrations for differentiation of cellulitis and deep vein thrombosis. AB - The differentiation between cellulitis and deep vein thrombosis was investigated in a series of patients with acutely swollen, painful, red legs. Oedema-fluid protein concentrations (as mean [SD]) were significantly higher in patients with cellulitis than in those with deep vein thrombosis (19.8 g/l [5.0] vs 5.85 g/l [1.6]), and the oncotic pressure of oedema-fluid was also significantly higher in cellulitis (6.3 mm Hg [1.3] vs 2.1 mm Hg [0.6]). In hospitals where phlebography, plethysmography, or doppler ultrasound are not available, measurement of oedema fluid protein concentrations may be a rapid and inexpensive guide to the cause of a red, swollen leg. PMID- 2570971 TI - Treatment of severe premenstrual syndrome with oestradiol patches and cyclical oral norethisterone. AB - 40 patients with premenstrual symptoms were randomly allocated to receive placebo patches or active treatment with transdermal oestradiol patches (2 x 100 micrograms) to suppress ovulation. Norethisterone 5 mg was given in each group from day 19-26 of the cycle to ensure a regular withdrawal bleed. Treatment was for 6 months with crossover at 3 months. Patients completed the Moos menstrual distress questionnaire (MDQ) and the premenstrual distress questionnaire (PDQ) daily throughout the study. 5 patients withdrew, 4 because of skin reactions and 1 because of considerable improvement with initial (active) treatment. After 3 months, both groups showed improvement in MDQ and PDQ scores. In general, between 3 and 6 months, patients who switched from active treatment to placebo had deteriorating scores while patients who switched from placebo to active treatment maintained or improved upon their initial gains. Significant improvements occurred after changing to active treatment in five of six negative MDQ symptom clusters and in six of ten PDQ symptoms. PMID- 2570972 TI - Clinical trials of zidovudine in HIV infection. PMID- 2570973 TI - Intermittent high-dose zidovudine. PMID- 2570974 TI - Inhibition of soluble CD4 therapy by antibodies to HIV. PMID- 2570975 TI - Psychosis and anti-tuberculosis therapy. PMID- 2570976 TI - Overt dialysis encephalopathy and mildly raised plasma aluminium. PMID- 2570977 TI - Plasma cholesterol, coronary heart disease, and cancer. PMID- 2570978 TI - School outbreak of gastroenteritis due to atypical rotavirus. PMID- 2570979 TI - APACHE-II score for assessment and monitoring of acute pancreatitis. PMID- 2570980 TI - Management of asthma in the community. PMID- 2570981 TI - Antibodies to Borrelia burgdorferi in Guillain-Barre syndrome. PMID- 2570982 TI - Back pain delaying diagnosis of metastatic testicular tumours. PMID- 2570984 TI - Rapid detection of beta s DNA from Guthrie cards by chromogenic probes. PMID- 2570983 TI - Preservatives in bronchodilator nebuliser solutions. PMID- 2570985 TI - Genetics of coronary heart disease. PMID- 2570986 TI - Mitral valve and wedge pressure. PMID- 2570987 TI - Gallstones, bile acids, and the liver. PMID- 2570988 TI - Overdiagnosis of infective endocarditis. PMID- 2570989 TI - Presenile dementia associated with mosaic trisomy 21 in a patient with a Down syndrome child. PMID- 2570990 TI - Informing patients about clinical disagreement. PMID- 2570991 TI - Rising age limit for kidney donors. PMID- 2570992 TI - Infants and very young children as kidney donors. PMID- 2570993 TI - University of Wisconsin preservation solute and bradyarrhythmia. PMID- 2570994 TI - Enhanced transcription of TNF in systemic vasculitis. PMID- 2570995 TI - Retinoic acid therapy for promyelocytic leukaemia. PMID- 2570996 TI - Modulation of coronary tone in acute myocardial infarction by endothelin. PMID- 2570997 TI - Endothelin and bronchial asthma. PMID- 2570998 TI - Sudden death and the Cambridge diet. PMID- 2570999 TI - Carboxyhaemoglobin and fatal methylene chloride poisoning. PMID- 2571000 TI - Severe chronic autoimmune haemolytic anaemia presenting haemolytic disease of the newborn. PMID- 2571001 TI - Vasculitis and retinoids. PMID- 2571002 TI - Hepatitis B insurance for the newborn. PMID- 2571003 TI - Squatting in second stage of labour. PMID- 2571004 TI - Are ACE inhibitors safe in pregnancy? PMID- 2571005 TI - Mumps vaccine and meningitis. PMID- 2571006 TI - Hispanic registry of graft procedures for Parkinson's disease. PMID- 2571007 TI - Automatism and the law. PMID- 2571008 TI - Latest AIDS figures. PMID- 2571009 TI - Effects of changes in fat, fish, and fibre intakes on death and myocardial reinfarction: diet and reinfarction trial (DART). AB - A randomised controlled trial with a factorial design was done to examine the effects of dietary intervention in the secondary prevention of myocardial infarction (MI). 2033 men who had recovered from MI were allocated to receive or not to receive advice on each of three dietary factors: a reduction in fat intake and an increase in the ratio of polyunsaturated to saturated fat, an increase in fatty fish intake, and an increase in cereal fibre intake. The advice on fat was not associated with any difference in mortality, perhaps because it produced only a small reduction (3-4%) in serum cholesterol. The subjects advised to eat fatty fish had a 29% reduction in 2 year all-cause mortality compared with those not so advised. This effect, which was significant, was not altered by adjusting for ten potential confounding factors. Subjects given fibre advice had a slightly higher mortality than other subjects (not significant). The 2 year incidence of reinfarction plus death from ischaemic heart disease was not significantly affected by any of the dietary regimens. A modest intake of fatty fish (two or three portions per week) may reduce mortality in men who have recovered from MI. PMID- 2571010 TI - Detection of a second widespread strain of Epstein-Barr virus. AB - The prevalence of Epstein-Barr virus (EBV) type B, which was previously found mainly in equatorial Africa, was investigated with the polymerase chain reaction in a population of healthy adults in Memphis, Tennessee. EBV was detected in the throat washings of 34 (22%) of 157 randomly selected donors, 14 (41%) of whom had type B virus and 17 (50%) type A; 3 donors (9%) had both strains. 18 additional adults with human immunodeficiency virus (HIV-1) infection and 6 severely immunocompromised children were also investigated. Results indicated that type B EBV is widespread in nature and may be reactivated by immunodeficiency. PMID- 2571012 TI - Percutaneous ultrasonic angioplasty:initial clinical experience. AB - Percutaneous catheter-delivered ultrasound energy for arterial recanalisation was applied in eight patients with peripheral vascular disease. Four patients had severe claudication with total occlusion and four high-grade stenosis in a superficial femoral or popliteal artery. A prototype ultrasound probe, with a frequency of 20 kHz and a power output of 20-35 W/cm2, was ensheathed in a 7F catheter and advanced to the occlusions under angiographic guidance. Three of four complete occlusions were recanalised in 120s or less, leaving a residual stenosis of mean (SD) diameter 54 (5)%. Ultrasound energy reduced the diameter of the isolated stenoses from 77 (14)% to 37 (21)%. All lesions were further treated with balloon angioplasty, resulting in a mean residual stenosis of 20 (9)%. There was no evidence of arterial emboli, dissection, spasm, or perforation. The major limitations of this ultrasound system are ease of steering and flexibility. Percutaneous catheter-delivered ultrasound energy appears promising for safe and effective use in atherosclerotic peripheral vessels to reduce arterial stenoses and recanalise complete arterial obstructions. PMID- 2571013 TI - HIV-associated enteropathy. PMID- 2571011 TI - Anal and cervical intraepithelial neoplasia: possible parallel. AB - This study evaluated prospectively the use of an endoscope in examination of the anal canal for the detection of premalignant lesions. All patients underwent endoscopy and anal epithelial biopsy; the biopsy samples were examined histologically and human papillomavirus (HPV) DNA hybridisation was done. No evidence of anal intraepithelial neoplasia (AIN) was found in 20 control patients with anal fissure or fistula. Of 82 patients with anal HPV infection, 23 had evidence of AIN. The prevalence of AIN was significantly higher among homosexual than among heterosexual men (17 of 28 vs 1 of 26) with anal HPV infection. Of 28 women with anal HPV infection, 10 had cervical intraepithelial neoplasia (CIN); 5 of the 10 also had AIN, whereas no woman had AIN in the absence of CIN. The study shows that AIN occurs and can be diagnosed endoscopically in a manner similar to CIN. Further detailed prospective studies on the natural history of AIN and of groups at risk are required. PMID- 2571014 TI - Homelessness. PMID- 2571015 TI - Topical NSAIDs: a gimmick or a godsend? PMID- 2571016 TI - Immunology of measles. PMID- 2571017 TI - Antibiotics for cholangitis. PMID- 2571018 TI - Heat injury and the armed forces. PMID- 2571019 TI - Upper gastrointestinal cancer in patients with familial adenomatous polyposis. AB - 102 patients with familial adenomatous polyposis underwent upper gastrointestinal endoscopy as a screening test for gastroduodenal adenomas. 100 had duodenal abnormalities (dysplasia in 94, and hyperplasia in 6), usually in the second and third parts of the duodenum (91%). The periampullary area was abnormal in 87 of 97 patients who had a biopsy specimen taken from this site (dysplasia 72, hyperplasia 13, and inflammation 2). By contrast, gastric dysplasia was found in only 6 patients. Classification of duodenal polyposis on a 5-grade scale (stages 0-IV), based on polyp number, size, histology, and severity of dysplasia, showed that 11 had stage IV disease: these patients are at greatest risk of malignant change and require close surveillance. The pattern of dysplasia observed in the upper gastrointestinal tract resembled the pattern of mucosal exposure to bile. PMID- 2571020 TI - Cell-surface peptidases as modulators of growth and differentiation. AB - Some of the many cell-surface antigens defined by the CD (cluster differentiation) nomenclature have lately emerged as proteins with well characterised enzymic activities. One important example is CD10 or CALLA (common acute lymphoblastic leukaemia antigen), which is identical to endopeptidase 24.11, an enzyme with an important role in the hydrolysis of biologically active peptides. CD13 and CD26 are also surface peptidases. These enzymes, which have a wide distribution on the surfaces of various cell types, may have specific roles in the control of growth and differentiation in both haemopoietic and epithelial cell systems. PMID- 2571021 TI - Gamma-hydroxybutyric acid for treatment of alcohol withdrawal syndrome. AB - The effect of gamma-hydroxybutyric acid (GHB) on ethanol withdrawal syndrome in alcoholics was investigated in a randomised double-blind study. Patients with withdrawal symptoms were treated either with GHB (orally in a syrup preparation) (11 patients) or with the syrup alone (12). GHB treatment (50 mg/kg) led to a prompt reduction in withdrawal symptoms, such as tremors, sweating, nausea, depression, anxiety, and restlessness. The only side-effect was dizziness. GHB may be useful in the management of alcohol withdrawal syndrome in man. PMID- 2571022 TI - Age of onset and type of leukaemia. PMID- 2571023 TI - Polychlorinated biphenyls: where do we stand now? PMID- 2571024 TI - The cost of treating AIDS. PMID- 2571025 TI - Bulimia while sleepwalking, a rebuttal for sane automatism? PMID- 2571026 TI - Antibodies to hepatitis C virus in French blood donors. PMID- 2571027 TI - Antibodies to hepatitis C virus in Italian blood donors. PMID- 2571028 TI - Liver disease in anti-HBe positive chronic HBsAg carriers and hepatitis C virus. PMID- 2571029 TI - Hepatitis C virus antibody. PMID- 2571030 TI - Antibodies to hepatitis C virus in non-A, non-B post-transfusion and cryptogenetic chronic liver disease. PMID- 2571031 TI - Melanoma risk assessment. PMID- 2571032 TI - Rapidly progressive Alzheimer's disease. PMID- 2571033 TI - Hyperbaric and normobaric oxygen in acute carbon monoxide poisoning. PMID- 2571034 TI - Aspirin and primary prophylaxis in myocardial infarction. PMID- 2571036 TI - Effect of salt restriction on hypertension. PMID- 2571035 TI - Prostaglandins and Campylobacter pylori. PMID- 2571037 TI - Use of apomorphine to test for dopamine responsiveness in Wilson's disease. PMID- 2571038 TI - Severe oedema after subcutaneous apomorphine in Parkinson's disease. PMID- 2571039 TI - OKT3-induced cytokine release attenuation by high-dose methylprednisolone. PMID- 2571040 TI - Heat illness in servicemen. PMID- 2571041 TI - Screening for congenital cytomegalovirus and toxoplasmosis. PMID- 2571042 TI - Stevens-Johnson syndrome due to pyrimethamine/sulfadoxine during presumptive self therapy of malaria. PMID- 2571043 TI - Leishmaniasis in France. PMID- 2571044 TI - Blood cell morphology in chronic fatigue syndrome. PMID- 2571045 TI - Development of chronic granulocytic leukaemia in the absence of a spleen. PMID- 2571046 TI - Cape crusaders. PMID- 2571047 TI - Declaration of vested interest. PMID- 2571048 TI - Forensic sciences in Israel. PMID- 2571049 TI - Occult cervical carcinoma revealed by large loop diathermy. PMID- 2571050 TI - Thoughts and ethics. PMID- 2571051 TI - Transient symptomatic hypotension in patients on simvastatin. PMID- 2571052 TI - Safety of amoxapine. PMID- 2571053 TI - Fatal toxic epidermal necrolysis associated with co-trimoxazole. PMID- 2571054 TI - Acute otitis media in very young infants. PMID- 2571055 TI - Nutritional supplements, stunting, and child development. PMID- 2571056 TI - Fish oil revisited. PMID- 2571057 TI - Unchanged serum lipoprotein (a) concentrations with lovastatin. PMID- 2571058 TI - Immune mechanisms in HIV-related neuropathies. PMID- 2571059 TI - Risk of HIV infection in polytransfused thalassaemia patients. PMID- 2571060 TI - Legal and moral responsibility for heatstroke in the armed forces. PMID- 2571061 TI - Effect of two formulations of a beta blocker on fibrinolytic response to maximum exercise. AB - It has been reported that catecholamine release is associated with stressful events and that adrenaline administration induces hyperfibrinolysis. To examine the possible involvement of the adrenoreceptor mechanism in the regulation of fibrinolytic activity at rest and to maximum exercise, 12 healthy volunteers (six males and six females, age range 19-21 yr) were orally medicated with two formulations of a beta blocker. The drug treatments consisted of 1) a single dose of OM756 alpha 10 mg propranolol "Inderal" tablet, 2) a single dose of a generic 10 mg propranolol tablet, and 3) a single dose of generic placebo. The drug administrations were assigned to the subjects using a complete Latin square design and double-blind procedure and were separated by 7 d. Two hours after the administration of the respective drug treatment, subjects exercised to maximal capacity using a graded exercise protocol on a bicycle ergometer. Prior to and 2 h after the drug administration and immediately after maximum exercise, heart rate, blood pressure, and oxygen consumption were measured and venous blood was removed and analyzed for fibrinolytic activity using a standard fibrin plate method. ANOVA showed that the resting and maximum heart rates were significantly reduced (P less than 0.05) following oral premedication with either of the two formulations of beta blocker, while blood pressure and oxygen consumption were unaffected. ANOVA also showed no difference in the resting fibrinolytic activity 2 h post-administration of the drug treatments but a significant decrease (P less than 0.05) in the normal fibrinolytic response to maximum exercise. It was concluded, therefore, that the resting level of fibrinolytic activity is not mediated via an adrenergic pathway but that the enhanced fibrinolytic activity to maximum exercise is partially regulated via an adrenoreceptor mechanism. PMID- 2571063 TI - A unified theory of mental illness. AB - I propose that in certain mental illnesses the mechanics for making, releasing, and responding to neurotransmitters are all normal; what is faulty is a portion of the mechanism whereby the brain recognizes abnormal behavior and feeds back on to the neuronal pathways producing the behavior to alter function of that neuronal system and re-establish normal behavior. All the above uses the physical structure of the brain. The existence of the feedback mechanism, and its perturbation, is deduced from data about the mechanism of antidepressants. PMID- 2571062 TI - Characterization of coated-vesicle adaptors: their reassembly with clathrin and with recycling receptors. AB - Adaptors sort out those receptors that participate in assembly of coated pits from those that are excluded. Two distinct adaptor units have so far been identified: (1) adaptors restricted to plasma membrane coated pits (HA-II type, named according to their elution position during hydroxylapatite chromatography) and (2) adaptors restricted to Golgi region coated pits (HA-I type). Adaptors contain a heterodimer of two 100-kDa polypeptides, a beta-adaptin (possibly carrying an essentially common clathrin-binding domain) and a distinct alpha- or gamma-adaptin characteristic of the type of adaptor and its specific location. Each adaptor in constructed from four different polypeptides. Thus HA-II adaptors contain a beta-adaptin and an alpha-adaptin in combination with a 50-kDa protein and a 16-kDa polypeptide. The HA-I adaptors contain a beta-adaptin and a gamma adaptin in combination with a 47-kDa protein and a 19-kDa polypeptide. Both types of adaptors and also a 180-kDa polypeptide will promote the assembly of clathrin to form coats, the size range of which appears to be relatively restricted compared to cages made from clathrin alone. The HA-II adaptors, characteristic of plasma membrane coated pits, bind to the cytoplasmic tail of the LDL receptor. They also assemble with the mannose 6-phosphate receptor in vitro in the absence of membrane. When clathrin is included, the adaptors promote the assembly of coats containing bound receptor. PMID- 2571064 TI - Pentamidine aerosol to prevent Pneumocystis carinii pneumonia. PMID- 2571066 TI - [Evaluation of the action of ivermectin on blood-sucking mosquitoes]. AB - The effect of Ivomec, a formulation of ivermectin administered by various routes: through biological membrane and feeder animal was assessed. Higher insecticidal effect was obtained by feeding mosquitoes with blood containing ivermectin through biological membrane. Anopheles stephensi females were the most sensitive of 3 species under study. Their total death after feeding was observed at 1 ppm, while for Aedes aegypti the lethal dose was 2.5 ppm and for An. sacharovi, 50 ppm. Subcutaneous injection of the drug to rabbits at the dose 0.34 mg/kg wt with further feeding on it of 3 Anopheles species demonstrated the highest drug sensitivity of An. stephensi mosquitoes. Death rates among the females fed on the rabbit 4-6 days after injection of the drug were 93, 70 and 79%, respectively. Death rates among An. sacharovi and An. atroparvus differed only slightly from that of the control group. In some experiments ivermectin effect on female fertility and larvae hatching from ova can be traced. Probably, increasing the preparation dose, administered to an animal-feeder, may change significantly characteristics of the malaria carrier natural population. Moreover, further use of ivermectin versus helminths and mosquitoes in zootechnical practice may produce a side-effect on mosquitoes, feeding recurrently on domestic animals, thus increasing the effect of malaria zoological control. PMID- 2571065 TI - [Morphofunctional changes of the ovarioles of blood-sucking mosquitoes (Diptera, Culicidae) during oogenesis. 2. Abortive oogenesis]. AB - The types of follicle's degeneration of 15 blood-sucking mosquito species were studied. Pregonotrophic degeneration takes place in 1-5 ovarioles immediately after imago hatching before feeding. Agonotrophic degeneration takes place in different number of ovarioles during long periods of sugar feeding. Gonotrophic degeneration takes place in some ovarioles at the same time with normal maturation of eggs in most of ovarioles after blood sucking. In ovarioles the relics are formed as a result of follicles' degeneration only and are named pregonotrophic, agonotrophic, gonotrophic relics, respectively. The possible causes of oosorption at different variants of follicles' degeneration are discussed. PMID- 2571067 TI - [The level and nature of insecticide resistance (DDT and malathion) in laboratory strain mosquitoes]. AB - Authors characterize 7 mosquito strains and species from the collection of the E. I. Martsinovskii Inst. with regard to the levels and mechanisms of their resistance to malathion and DDT. Resistance mechanisms of the strains An. stephensi, An. sacharovi, An. atroparvus, Ae. aegypti and C. pipiens were determined with the help of synergists. Malathion-resistance in one of the An. stephensi strains is due to high carboxyl activity. Other strains featured various levels of DDT-resistance due to various mechanisms: in Ae. aegypti it was due to mixed-function oxidases activity; in An. atroparvus, An. sacharovi and An. stephensi second strain to glutathione-dependent transferase, in C. pipiens, An stephensi third strain and, partially, An. sacharovi,--to the probable presence of kdr-factor. PMID- 2571068 TI - [Blood-sucking mosquitoes (Diptera, Culicidae) in the northern part of Udmurtia]. AB - Breeding sites and number of man-attacking mosquitos were studied in 1986-1987 in recreation areas in the north of Udmurt ASSR. Some sanitary-prophylaxis measures are recommended basing on the knowledge of breeding and activity periods of man biting mosquitos. Studies of the probable breeding sites of blood-sucking mosquitos were performed in 1986-1987 in the vicinity of pioneer camps "Zvezdochka", "Lastochka", the tourist base "Cheptsa" and Institute's sports camp in the north of the Udmurt ASSR. It was established that water basins in the floodlands of the Cheptsa River and its confluents, featuring rising water level during the floods, were the largest breeding sites of mosquitos. The most numerous species of man-biting mosquitos were Ae. cantans, Ae. punctor, the most common ones were Ae. excrucians, Ae. cinereus and Ae. vexans. Most of the mosquitos attacked in the second half of June--first half of August. PMID- 2571070 TI - Isolation of the facA (acetyl-coenzyme A synthetase) and acuE (malate synthase) genes of Aspergillus nidulans. AB - Acetate inducible genes of Aspergillus nidulans were cloned via differential hybridization to cDNA probes. Using transformation of mutant strains the genes were identified as facA (acetyl-Coenzyme A synthetase) and acuE (malate synthase). The levels of RNA encoded by these genes were shown to be acetate inducible and subject to carbon catabolite repression. Induction is abolished in a facB mutant and carbon catabolite repression is relieved in a creA mutant. PMID- 2571069 TI - Molecular and genetic analysis of an embryonic gene, DC 8, from Daucus carota L. AB - To understand the morphogenetic and physiological processes occurring during plant embryogenesis, we isolated cDNA clones homologous to genes preferentially expressed during somatic embryogenesis. One of these cDNA clones detected an embryo-specific mRNA species with a corresponding protein of 66 kDa. The expression pattern of the mRNA is similar between somatic and zygotic embryos of carrots. To characterize the gene encoding this mRNA, we isolated the corresponding genomic clones. Molecular analysis of the DNA from several haploid and diploid carrots showed that the mRNA was encoded by a single copy gene, named DC 8. DNA sequence analysis showed that the gene consisted of three exons and coded for a hydrophilic protein with a central region composed of 17 repeats. At the NH2-terminus no typical signal sequence was found. Immunocytochemical analysis localized the protein primarily in the vacuoles and protein bodies of zygotic embryos; the cytoplasm showed some antibody staining. The protein was also found in cell walls of endosperm tissue. The amount of DC 8 protein was too low for it to be categorized as a seed storage protein; its role in embryo genesis remains to be determined. PMID- 2571072 TI - Binding of vinblastine to stabilized microtubules. AB - Addition of 2-200 microM vinblastine to microtubules at steady state in vitro causes the microtubules to depolymerize, with the formation of protofilament spirals and other aggregated forms of microtubule protein. The presence of such spirals and protein aggregates, which are difficult to separate from microtubules, has complicated attempts to measure the binding of vinblastine to microtubules. We have found that stabilizing bovine brain microtubules in vitro with dimethyl sulfoxide, taxol, or a combination of dimethyl sulfoxide and taxol prevents or greatly retards the formation of protofilamentous spirals, thus permitting us to measure the binding of vinblastine to intact microtubules. Reciprocal plots of binding data indicate the presence of 1.4-1.7 vinblastine binding sites/mol of tubulin in the microtubule, with a Ka of approximately 3-4 x 10(3) M-1. The Ka value obtained is within 1 order of magnitude of the apparent intrinsic binding constant for the binding of vinblastine to tubulin dimers. The results support the idea that depolymerization of microtubules by intermediate and high concentrations of vinblastine occurs by stoichiometric binding of vinblastine to tubulin along the microtubule surface. PMID- 2571071 TI - 1-beta-D-arabinofuranosylcytosine enhancement of resistance to several antineoplastic drugs in mammalian tissue culture cells. AB - This report describes the enhancement of drug resistance by 1-beta-D arabinofuranosylcytosine at four individual loci and gene amplification at one locus in a hamster cell line. This drug has been used chemotherapeutically in the treatment of neoplasia and has documented effects on DNA synthesis. Our results in this paper demonstrate that the incidence of resistance to methotrexate, N (phosphonoacetyl)-L-aspartate, and 5-fluoro-2'-deoxyuridine was appreciably increased after pretreatment with 1-beta-D-arabinofuranosylcytosine and they show that an increase in the incidence of gene amplification is one of the mechanisms by which drug resistance is increased. In contrast, the incidence of vincristine resistance was minimally enhanced by this drug. Possible reasons for this differential enhancement are discussed. PMID- 2571073 TI - Functional characterization of a rat dopamine D-2 receptor cDNA expressed in a mammalian cell line. AB - We recently cloned a complementary DNA for the rat dopamine D-2 receptor, making it possible to create cell lines expressing this receptor. A cell line (LZR1) was created by transfecting the D-2 cDNA (RGB-2) into mouse fibroblast Ltk- cells. LZR1 cells, previously described as L-RGB2Zem-1 cells, express a high density of D-2 receptors, whereas the wild-type cells do not. A number of agonists competitively and stereoselectively inhibited the binding of [3H]spiroperidol to the expressed D-2 receptors in a GTP-sensitive manner. The potency of dopamine was decreased by the addition of GTP. NaCl and GTP together caused a further decrease in potency and increased the Hill slope for inhibition of radioligand binding by dopamine almost to 1.0. Pretreatment of cells with pertussis toxin inhibited high affinity binding of dopamine and prevented further inhibition of binding by GTP. The NaCl-induced decrease in affinity was not prevented by pertussis toxin treatment. Dopamine reduced forskolin-stimulated adenylate cyclase activity by 27% in membranes prepared from LZR1 cells. Inhibition by dopamine was blocked by (+)-butaclamol or prior treatment of intact cells with pertussis toxin. Other dopamine receptor agonists stereoselectively inhibited adenylate cyclase activity. These data indicate that the RGB-2 cDNA directs the expression of a dopamine D-2 receptor capable of interacting with guanine nucleotide-binding proteins and inhibiting adenylate cyclase activity. Furthermore, the RGB-2 cDNA provides a means of creating many cell lines that will be useful tools for the biochemical and pharmacological characterization of dopamine D-2 receptors. PMID- 2571074 TI - Effect of taxol on the heparin-induced secretion of lipoprotein lipase from cardiac myocytes. AB - The heparin-induced secretion of LPL into the incubation medium of cardiac myocytes occurred in two phases: a rapid release (5-10 min), followed by a slower rate of release (10-60 min). Reducing the incubation temperature from 37 degrees C to 23 degrees C inhibited the slow phase of secretion, but had no effect on the rapid phase. Similarly, taxol, a microtubule-stabilizing drug, selectively reduced the slow phase of LPL release, without influencing the rapid release of LPL into the medium or cellular LPL activity. The rapid heparin-induced release of LPL probably occurs from sites that are at or near the cell surface, and so microtubules must participate in the intracellular transport of LPL from sites of synthesis and glycosylation to the surface binding sites. Heparin-releasable LPL could be resolved into two fractions by chromatography on con A-Sepharose; this pattern of elution was not affected by the prior treatment of cardiac myocytes with taxol. PMID- 2571075 TI - Biochemical characterization of chromatin fractions isolated from induced and uninduced Friend erythroleukemia cells. AB - Chromatin fractions from Friend erythroleukemia cells after induction of differentiation by dimethylsulfoxide (DMSO) were compared in their biochemical characteristics to fractions from uninduced cells. Fractions were prepared by extracting chromatin from nuclei after mild micrococcal nuclease treatment with increasing concentrations of NaCl according to Sanders. This procedure has been found to release chromatin containing hyperacetylated histones preferentially. The fractions obtained by this procedure were analysed in respect to the amount of chromatin released, the amount of histone H1, the degree of acetylation of histone H4, the presence of non-histone proteins and the concentration of transcribed and non-transcribed sequences. It was found that the fractions differ in the amount of histone H1 present, in several non-histone proteins and in the acetylation of histone H4, regardless whether induced or uninduced cells were analysed. The distribution of transcribed sequences versus non-transcribed sequences among the fractions was the same, demonstrating that this fractionation procedure, although leading to fractions with biochemical differences, is not able to discriminate functional states of chromatin and that the biochemical characteristics of the fractions may be common to both, active as well as inactive states of chromatin. PMID- 2571076 TI - Hormonal regulation of TSE1-repressed genes: evidence for multiple genetic controls in extinction. AB - Somatic cell hybrids formed by fusing hepatoma cells with fibroblasts generally fail to express liver functions, a phenomenon termed extinction. Previous studies demonstrated that extinction of the genes encoding tyrosine aminotransferase, phosphoenolpyruvate carboxykinase, and argininosuccinate synthetase is mediated by a specific genetic locus (TSE1) that maps to mouse chromosome 11 and human chromosome 17. In this report, we show that full repression of these genes requires a genetic factor in addition to TSE1. This conclusion is based on the observation that residual gene activity was apparent in monochromosomal hybrids retaining human TSE1 but not in complex hybrids retaining many fibroblast chromosomes. Furthermore, TSE1-repressed genes were hormone inducible, whereas fully extinguished genes were not. Analysis of hybrid segregants indicated that genetic loci required for the complete repression phenotype were distinct from TSE1. PMID- 2571077 TI - DNA methylation patterns associated with asparagine synthetase expression in asparagine-overproducing and -auxotrophic cells. AB - In Chinese hamster ovary cells, the gene for asparagine synthetase, which spans 20 kilobase pairs, was found to contain a cluster of potential sites for CpG methylation in a 1-kilobase-pair region surrounding the first exon. Fourteen of the sites that could be assayed for methylation by MspI-HpaII digestions were found in this region, with an additional nine MspI sites spread throughout the remainder of the gene. The methylation status of the gene was analyzed in a series of cell lines that differed in the amount of asparagine synthetase activity. The level of expression showed a direct correlation with the extent of methylation of a subset of the MspI sites found in the 5' region of the gene. The rest of the gene was completely methylated in most cell lines. Wild-type cells, which expressed a basal level of asparagine synthetase activity, were partially demethylated in the 5' region. In contrast, asparagine-requiring N3 cells, which lacked detectable mRNA for asparagine synthetase, were methylated throughout the entire gene. Spontaneous revertants of strain N3, selected for growth in asparagine-free medium, exhibited extensive hypomethylation of the asparagine synthetase gene. The methylation pattern of the gene in cell lines that overproduced the enzyme was also examined. Albizziin-resistant cell lines, which had amplified copies of the gene, were extensively demethylated in the 5' region. Overexpression of asparagine synthetase in beta-aspartyl hydroxamate-resistant lines without amplified copies of the gene was also correlated with DNA hypomethylation. PMID- 2571079 TI - Translational status of proenkephalin mRNA in the rat reproductive system. AB - The mRNA for the opioid peptide precursor proenkephalin is widely distributed throughout the male and female reproductive systems of rodents. In the present studies, the concentrations of proenkephalin-derived peptides in selected reproductive tissues of the rat have been determined. When compared with previously characterized tissues such as brain, the peptide contents in reproductive tissues were unexpectedly low relative to the abundance of proenkephalin mRNA. This suggested that either translation of proenkephalin mRNA is relatively inefficient in reproductive tissues or that the turnover of proenkephalin products occurs at a higher rate, or both. To distinguish between these possible mechanisms, the polysomal distributions of proenkephalin mRNA in different rat reproductive tissues and in rat brain were determined. In adult rat testis, in which the predominant proenkephalin RNA is the 1700-nucleotide form present in spermatogenic cells, the transcript was found to be mainly associated with translationally inactive ribonucleoprotein fractions. In contrast, the 1450 nucleotide form of proenkephalin mRNA appeared to be translated to a similar extent in rat brain, epididymis, ovary, and somatic cells of the immature rat testis. It therefore appears that inefficient translation of proenkephalin mRNA in spermatogenic cells is a major determinant of the low ratio of proenkephalin peptides to RNA in the adult rat testis, while posttranslational mechanisms (most likely peptide turnover) are involved in the rat epididymis, ovary, and presumably other reproductive tissues. These findings also indicate that mRNA and/or translation product concentrations within a given tissue are not always accurate indicators of the level of peptide or protein production. PMID- 2571081 TI - Evidence of pre-prosomatostatin mRNA in human normal and tumoral anterior pituitary gland. AB - Expression of the SRIH gene was investigated in six human normal anterior pituitaries, six GH-, three PRL-, three mixed GH/PRL-secreting and four nonsecreting adenomas. Total cellular RNA and poly(A+) mRNAs were analyzed by dot and Northern blot hybridization to a 3'-end labeled oligonucleotide probe specific for the human pre-proSRIH mRNA. A weak but detectable pre-proSRIH hybridization signal was present in human normal anterior pituitaries and in the four groups of adenomas. The size of this pre-proSRIH mRNA was indistinguishable from that found in our hypothalamic samples and close to that described in the literature. The wide variation of the signal intensity from one case to the other in each group of the different types of normal and tumoral antehypophyseal samples prevented establishment of any correlation between the level of pre proSRIH mRNA and the nature of the pituitary tissue. The presence of SRIH mRNA in human normal and tumoral anterior pituitary tissues provides a sound basis to substantiate the hypothesis of a SRIH biosynthesis in the human anterior pituitary gland. PMID- 2571078 TI - Structure and expression of the human MDR (P-glycoprotein) gene family. AB - The human MDR (P-glycoprotein) gene family is known to include two members, MDR1 and MDR2. The product of the MDR1 gene, which is responsible for resistance to different cytotoxic drugs (multidrug resistance), appears to serve as an energy dependent efflux pump for various lipophilic compounds. The function of the MDR2 gene remains unknown. We have examined the structure of the human MDR gene family by Southern hybridization of DNA from different multidrug-resistant cell lines with subfragments of MDR1 cDNA and by cloning and sequencing of genomic fragments. We have found no evidence for any other cross-hybridizing MDR genes. The sequence of two exons of the MDR2 gene was determined from genomic clones. Hybridization with single-exon probes showed that the human MDR1 gene is closely related to two genes in mouse and hamster DNA, whereas MDR2 corresponds to one rodent gene. The human MDR locus was mapped by field-inversion gel electrophoresis, and both MDR genes were found to be linked within 330 kilobases. The expression patterns of the human MDR genes were examined by enzymatic amplification of cDNA. In multidrug-resistant cell lines, increased expression of MDR1 mRNA was paralleled by a smaller increase in the levels of MDR2 mRNA. In normal human tissues, MDR2 was coexpressed with MDR1 in the liver, kidney, adrenal gland, and spleen. MDR1 expression was also detected in colon, lung, stomach, esophagus, muscle, breast, and bladder. PMID- 2571080 TI - Expression of low density lipoprotein receptor gene in human placenta during pregnancy. AB - Mammalian cells require cholesterol as a structural component of plasma membranes. It is also required for placental steroid synthesis. De novo synthesis of cholesterol is limited in human placenta and cholesterol is obtained mainly from plasma low density lipoprotein (LDL). Cholesterol delivery from LDL is mediated by receptor-mediated uptake and the receptor amount is the most important factor for cellular delivery. Thus, the regulation of receptor synthesis is important for placental development and function. Since the regulation of LDL receptor gene expression has not been studied in human placenta, LDL receptor mRNA was measured in placentae of 5-40 weeks of gestation by hybridization of RNA with 32P-labeled cDNA for human LDL receptor. Two mRNA species for LDL receptor were demonstrated by Northern blot analysis. The longer mRNA [5.3 kilobases (kb)] was much more abundant than the shorter mRNA (3.7 kb). The amount of 5.3 kb mRNA was highest early in gestation and decreased during pregnancy. However, the amount of 3.7 kb mRNA did not change appreciably during gestation. Dot blot analysis of 26 placental mRNAs obtained from various stages of gestation revealed a negative correlation between LDL receptor mRNA and gestation (r = -0.76, P less than 0.001). Considering the rapid growth of the trophoblast during gestation, especially in the first and the second trimester, increased expression of the LDL receptor gene and subsequent translation are expected for efficient cholesterol uptake to provide a sufficient substrate for cell growth. Possible mechanisms for the appearance of two mRNA species for LDL receptor are also discussed. PMID- 2571082 TI - Antiparkinsonian activity of CY 208-243, a partial D-1 dopamine receptor agonist, in MPTP-treated marmosets and patients with Parkinson's disease. AB - The effect of stimulation of cerebral dopamine D-1 receptors by CY 208-243 on motor disability was tested in MPTP-treated parkinsonian marmosets and patients with Parkinson's disease. CY 208-243 (0.5-1.25 mg/kg s.c.) produced a dose related reversal of akinesia and rigidity in the marmosets, lasting some 2 h. Single morning doses of CY 208-243 (5-40 mg) were compared with the usual morning dose of levodopa in eight patients with Parkinson's disease on long-term levodopa therapy who had developed motor fluctuations from immobility with akinesia and rigidity (off) to mobility often with dyskinesias (on). CY 208-243 alone was capable of switching such patients from off to on; five of the eight patients responded to the highest dose (40 mg), sometimes with dyskinesias. The response to CY 208-243 was comparable to that produced by levodopa in these cases. Drugs designed to stimulate both dopamine D1 and D2 receptors in the brain may improve the therapy of Parkinson's disease. PMID- 2571083 TI - CQP 201-403 in Parkinson's disease: an open-label pilot study. AB - The dopamine agonist, CQP 201-403, was administered to 10 patients in an open label fashion with rapid dosage escalation during hospitalization. Assessed over an average of 20 days, significant improvement occurred in bradykinesia, rigidity, and postural instability. Tremor did not occur in sufficient frequency in this group of patients to be accurately assessed. The most serious adverse effect encountered was prolonged confusion with psychosis. This study suggests that CQP 201-403 may be of value in the treatment of Parkinson's disease. PMID- 2571084 TI - Clinical implications of positive tests for antibodies to human immunodeficiency virus type 1 in asymptomatic blood donors. AB - Of 693,000 volunteer blood donors in Washington, D.C., who were screened for infection with human immunodeficiency virus type 1 (HIV-1) from July 1985 through December 1988, 284 tested positive on both enzyme immunoassay and Western blot assay. To determine the clinical importance of confirmed positive test results in asymptomatic blood donors, we followed 156 donors with positive Western blot assays and 80 donors with positive enzyme immunoassays but negative or indeterminate Western blots at 6-month intervals for a mean of 28 months. As compared with Western blot-negative persons, those with positive Western blots were significantly more likely to be black, male, and first-time donors and to have a history of venereal disease, generalized lymphadenopathy on examination, CD4-cell counts lower than 0.4 x 10(9) per liter, IgG levels higher than 18 g per liter, and antibody to hepatitis B core antigen on initial evaluation. In 17 (11 percent) of the Western blot-positive donors, the disease progressed to Class IV (symptomatic disease), according to the Centers for Disease Control system. CD4 counts below 0.2 x 10(9) per liter, IgA levels above 4 g per liter, abnormal proliferative responses to tetanus toxoid, and positive viral cultures were the strongest predictors of disease progression. Among the 80 donors with repeatedly reactive assay results but either negative or indeterminate Western blot assays, there was no evidence of HIV exposure in their histories, physical examinations, or laboratory evaluations, and manifestations of HIV infection developed in none of them. We conclude that a small number of persons with HIV infection continue to donate blood, despite attempts to exclude them, but that donors who test positive on enzyme immunoassay but persistently negative or indeterminate on Western blot assay probably do not represent a risk for the transmission of HIV. PMID- 2571085 TI - The molecular biology of Laron dwarfism and medullary thyroid cancer. PMID- 2571087 TI - Segment-specific expression of a homoeobox-containing gene in the mouse hindbrain. AB - The process of segmentation, in which the developing embryo is divided into repetitive structures along its antero-posterior (A-P) axis, as a means of organizing and coordinating the body plan is found in a wide range of organisms. In Drosophila, homoeotic genes are involved in all levels of segmental organization and in determining segment identity. The roles of these genes in segmentation have been found mainly by mutational studies, but also by in situ hybridization, which has shown their domains of expression. In contrast to Drosophila, however, embryonic expression of homoeobox-containing genes in vertebrate organisms has not been found to follow a segmental pattern. Vertebrate segmentation can be clearly seen in the mesodermal somites, but repetitive morphological structures in the central nervous system (neuromeres) have only recently been shown to have developmental significance. Neuromeres in the hindbrain (rhombomeres) have been defined as segmental units by their pattern of nerve formation in the developing chick and by the alternating expression of Krox 20, a gene encoding a zinc-finger DNA-binding protein, in the 9.5-day-old mouse. Here we report that a mouse homoeobox-containing gene, Hox-2.9, is expressed in a segment-specific manner in the developing mouse hindbrain. This expression is in a region which is flanked by the regions of expression of Krox-20, and is precisely contained within a single neuromere, rhombomere 4. PMID- 2571086 TI - Screening for multiple endocrine neoplasia type 2a with DNA-polymorphism analysis. AB - Multiple endocrine neoplasia type 2a has been shown to be genetically linked to a locus near the centromere of chromosome 10. The availability of polymorphic DNA probes for the region permits the use of restriction-fragment-length polymorphisms (RFLP) to identify carriers of the gene for this cancer syndrome. As part of a French national program, DNA probes were used in a genetic-linkage study of 130 members of 11 families of European and North African origin. In these families there was no recombination between the mutation causing multiple endocrine neoplasia type 2a and two of the three probes used. All 11 families were informative for at least one of the three markers, and linkage information was adequate to provide genetic counseling to 8 families. We found that RFLP analysis is much more useful in predicting the carrier state than conventional endocrine challenge, especially in younger people, but accuracy is maximal when both methods are employed. We conclude that genetic screening allows the identification of those who are at risk for multiple endocrine neoplasia type 2a at any age with a high level of certainty. After initial screening with DNA, tests for early neoplastic change may be directed toward those determined to be at high risk. PMID- 2571088 TI - AIDS treatment. New yardstick for success. PMID- 2571089 TI - DNA forensics and the FBI. PMID- 2571090 TI - NMDA and non-NMDA receptors are co-localized at individual excitatory synapses in cultured rat hippocampus. AB - A CENTRAL assumption about long-term potentiation in the hippocampus is that the two classes of glutamate-receptor ion channel, the N-methyl-D-aspartate (NMDA) and the kainate/quisqualate (non-NMDA) subtypes, are co-localized at individual excitatory synapses. This assumption is important because of the perceived interplay between NMDA and non-NMDA receptors in the induction and expression of long-term potentiation: the NMDA class, by virtue of its voltage-dependent channel block by magnesium and calcium permeability, provides the trigger for the induction of long-term potentiation, whereas the actual enhancement of synaptic efficacy is thought to be provided by the non-NMDA class. If both receptor subtypes are present at the one synapse, such cross-modulation could occur rapidly and locally through diffusible factors. By measuring miniature synaptic currents in cultured hippocampal neurons we show that the majority (approximately 70%) of the excitatory synapses on a postsynaptic cell possess both kinds of receptor, although to different extents. Of the remaining excitatory synapses, approximately 20% contain only the non-NMDA subtype and the rest possess only NMDA receptors. This finding provides direct evidence for co-localization of glutamate-receptor subtypes at individual synapses, and also points to the possibility that long-term potentiation might be differentially expressed at each synapse according to the mix of receptor subtypes at that synapse. PMID- 2571091 TI - Caenorhabditis elegans has scores of homoeobox-containing genes. AB - Homoeobox-containing genes control cell identities in particular spatial domains, cell lineages, or cell types during the development of Drosophila and Caenorhabditis elegans, and they probably control similar processes in vertebrates. More than 80 genes with homoeoboxes that have sequence similarities ranging from 25 to 100% have been isolated by genetic means or by DNA hybridization to previously isolated genes. We synthesized 500-2,000-fold degenerate oligonucleotides corresponding to a set of well-conserved eight amino acid sequences from the helix-3 region of the homoeodomain. We screened C. elegans genomic libraries with these probes and identified 49 putative homoeobox containing loci. DNA sequencing confirmed that eight out of ten selected loci had sequences corresponding to the conserved helix-3 region plus additional flanking sequence similarity. One of these genes contained a sequence corresponding to a complete pou-domain and another was closely related to the homoeobox-containing genes caudal/cdx-1. The putative homoeobox loci were mapped to the physical contig map of C. elegans, allowing the identification of potentially corresponding genes from the correlated genetic map. We estimate that the number of homoeobox-containing genes in C. elegans is at least 60, constituting approximately 1% of the estimated total number of genes. PMID- 2571092 TI - A sporadic case of acute Hantavirus nephropathy in The Netherlands. AB - A 32-yr-old man presented with acute renal failure preceded by a viral-like disease with high fever, bilateral loin pain, nausea, headache and slight thrombocytopenia. Renal biopsy revealed only minor tubulointerstitial abnormalities. Renal function completely normalised within 10 days after hospitalisation. The diagnosis of acute Hantavirus nephropathy was proved by serological examinations. PMID- 2571093 TI - Changes in regional levels of putative neurotransmitter amino acids in brain under unilateral forebrain ischemia. AB - The levels of the neurotransmitter amino acids glutamate, aspartate, and GABA were determined in different brain regions during ischemia and post-ischemic recirculation periods using the unilateral carotid artery occlusion model of stroke in gerbils. The levels of glutamate, aspartate and GABA in ischemic hemisphere were increased significantly by 10 min of ischemia and later declined with time. Reperfusion for 30 min following 10 min. of ischemia further enhanced the levels of glutamate and aspartate. Increase in GABA levels were found during early periods of reperfusion. Regional variations in the changes of amino acids' levels were noticed following ischemia. Hippocampus showed the highest increase in glutamate levels followed by striatum and cerebral cortex. Aspartate levels in striatum and hippocampus increased during 10 min ischemia (46% and 30%) and recirculation (70% and 79%), whereas in cerebral cortex the levels were doubled only during recirculation. Ischemia induced elevations of GABA levels were observed in cerebral cortex (68%) and in hippocampus (30%), and the levels were normalized during recirculation. No changes in GABA levels were found in striatum. It is suggested that the large increase in the levels of excitatory neurotransmitter amino acids in brain regions specially in hippocampus during ischemia and recirculation may be one of the causal factors for ischemic brain damage. PMID- 2571094 TI - Measurement of amino acid release from cultured cerebellar granule cells by an improved high performance liquid chromatography procedure. AB - Endogenous amino acid release was examined in rat cerebellar primary cultures comprising more than 95% of glutamatergic granule cells. Eighteen amino acids were determined in the cell extracts and in the release fractions by high performance liquid chromatography, using precolumn derivatization with o phthaldialdehyde and separation on a reverse-phase column using a multi-step gradient system of two solvents (0.1 M Na+ acetate, pH 7.2/methanol: tetrahydrofuran, 97:3). The fluorimetric response was linear, at least in the range of 2-162 pmol, for all the amino acids analysed, with a detection limit of 1 pmole. We observed a good reproducibility in within-assay and between-assay coefficients of variation of the retention times and fluorescence yield. When cultured granule cells were exposed to the excitatory amino acid receptor agonist quisqualic acid (50 microM), we observed a net increase in the release of glutamate (3 fold over the baseline) and a smaller increase in that of aspartate (2 fold) and taurine (1.6 fold). Other amino acids were not significantly affected. GABA levels were below detection limits, due to the minimal number of GABAergic neurons present in the cultures. PMID- 2571095 TI - Beta-DL-methylene-aspartate, an inhibitor of aspartate aminotransferase, potently inhibits L-glutamate uptake into astrocytes. AB - [3H]Glutamate uptake into astrocytes in primary culture was potently inhibited by the aspartate analogues L- and D-aspartic acid, DL-threo-beta-hydroxy-aspartic acid-beta-hydroxymate (IC50's: 136, 259, 168, and 560 microM, respectively) and by beta-DL-methylene-aspartate, a suicide inhibitor of aspartate aminotransferase (IC50: 524 microM), and by the endogenous sulphur-containing amino acid L cysteinesulfinic acid (IC50: 114 microM), [3H]Glutamate uptake was not significantly affected by either N-methyl-D-aspartate or DL-homocysteine thiolactone. These results demonstrate that other excitatory amino acids including aspartate and L-cysteinesulfinic acid (but excluding L-homocysteic acid) interact with the glutamate transport system of astrocytes. Inhibition of glutamate uptake may significantly increase the level of neuronal excitability. PMID- 2571096 TI - Lack of effect of chronic desipramine treatment on dopaminergic activity in the nucleus accumbens of the rat. AB - The binding of [3H]SCH 23390 to dopamine (DA) D1-receptors was measured in the nucleus accumbens of rats treated chronically with desipramine for 14 days. DA D1 and D2-receptor binding using [3H]SCH 23390 and [3H]spiperone, respectively as ligands, was determined in rats treated for 28 days. Neither Bmax nor Kd values were influenced by chronic desipramine treatment. In addition, chronic desipramine treatment (28 days) did not influence the dose dependent, quinpirole (10-1000 nM)-mediated inhibition of the electrically stimulated release of [3H]DA release and [14C]ACh from nucleus accumbens slices or the dose dependent increase in [3H]DA release and decrease in [14C]ACh release in the presence of 1 and 10 microM nomifensine. Therefore, our results suggest that the effect of chronic antidepressant treatment cannot be attributed to changes in either DA D1- or D2 receptor binding or DA D2-receptor function in the nucleus accumbens. PMID- 2571097 TI - Lack of excitatory amino acid-induced effects on calcium fluxes measured with 45Ca2+ in rat cerebral cortex synaptosomes. AB - Ca2+ uptake was measured in purified rat cerebral cortex synaptosomes (P3 pellets) using 45Ca2+ as a tracer. Ca2+ influx increased in time, and with an increase in external K+ concentration and temperature. The net (external K+ induced, depolarization-dependent) uptake follows a two-component course. The exponential term, due to the opening of voltage-operated calcium channels (VOC), has a rate constant which increases with an increase in the depolarization level (1.04 versus 0.54 nmol/s/mg protein for 50 mM - versus 15 mM [K+]-dependent net influx). The linear term, due to the Na+/Ca2+ exchange system, has a similar rate constant at all depolarization levels (0.16 +/- 0.05 and 0.11 +/- 0.02 nmol/s/mg protein). Excitatory amino acids (glutamate, kainate and n-methyl-d-aspartate NMDA-) were tested on this preparation at doses ranging between 5 x 10(-5) M and 5 x 10(-3) M and at multiple incubation times, under resting conditions and under two depolarizing conditions (partial depolarization: 15 mM external K+ and maximal depolarization: 50 mM external K+). NMDA was also tested in the absence of Mg2+. No effect was detectable under any of these experimental conditions. Hypotheses to interpret these data are discussed. Further studies on other preparations are needed in order to directly investigate the presynaptic effects of excitatory amino acids. PMID- 2571099 TI - Physiological role of somatostatin-mediated autofeedback regulation for growth hormone: importance of growth hormone in triggering somatostatin release during a trough period of pulsatile growth hormone release in conscious male rats. AB - In mammals including human, it is generally accepted that growth hormone (GH) can regulate its own secretion through an autofeedback mechanism in which somatostatin (SRIF) may be involved. To explore a physiological role of SRIF mediated GH autoregulation, the effect of exogenous human GH administration on plasma rat GH response to [D-Ala2, Nle27]-human GH-releasing hormone-(1-28) agmatine (hGHRH-analog), which does not crossreact with anti-rat GH-releasing hormone gamma-globulin (GHRH-Ab), was examined in conscious male rats treated with GHRH-Ab in the absence and presence of anti-SRIF gamma-globulin (SRIF-Ab). Enhanced SRIF release during a trough period of natural pulsatile GH secretion, suggested by the blunted GH response to exogenous hGHRH-analog, no longer occurred when major GH secretory bursts were abolished by GHRH-Ab treatment. On the other hand, when hGH was administered in GHRH-Ab-treated rats so as to simulate the quantity and dynamic change of GH in hypophysial portal circulation in rats exhibiting pulsatile GH secretion, hGHRH-analog-induced GH rises were significantly suppressed during the period corresponding to a GH trough. This suppression was completely prevented by simultaneous treatment with SRIF-Ab. Furthermore, administration of bovine GH, but not ovine prolactin, resulted in significant suppression of hGHRH-analog-provoked GH rises. These findings suggest that enhanced SRIF release during a trough period of spontaneous GH secretory rhythm is induced by the preceding GH secretory burst, and also suggest a possible role for SRIF-mediated GH autoregulation in a physiological state. PMID- 2571098 TI - Effect of histamine on the development of astroglial cells in culture. AB - The effect of histamine on different aspects of the growth of astrocytes was studied using primary cultures derived either from forebrain or from cerebellum of the rat. The influence on general growth and differentiation was monitored in terms of the activities of ornithine decarboxylase and glutamine synthetase enzymes, whereas [3H]thymidine incorporation into DNA was used as a specific index of cell proliferation. Treatment with 500 nM histamine of cells grown for 6 days in vitro, caused a time-dependent significant increase in ornithine decarboxylase activity of astrocytes from both sources. The maximum increase was observed at 4 h after histamine treatment, at that time the elevation in ornithine decarboxylase activity being about 80% and 300% over control values in the forebrain and the cerebellar astrocytes, respectively. Under similar experimental conditions, addition of histamine (500 nM) to medium resulted in a significant increase in [3H]thymidine incorporation into DNA in both types of cultures: in comparison with control, the elevation was about 45% at 48 h in forebrain astrocytes and at 24 h in cerebellar astrocytes. On the other hand, the specific activity of glutamine synthetase in cerebellar astrocytes was markedly enhanced (about 100%) by treatment with histamine (500 nM) for 4 days, but forebrain astrocytes were little affected. Addition of histamine to the culture medium produced no significant alteration in the activity of lactate dehydrogenase and protein content of either type of astroglial cells. The present findings, which support our earlier proposal that the biochemical properties of astrocytes differ between various brain regions, provide direct evidence for the involvement of histamine in the regulation of growth and development of astrocytes. PMID- 2571100 TI - Effects of the sulphydryl inhibitor N-ethyl-maleimide on the phrenic nerve and diaphragm muscle of the rat. AB - N-Ethyl-maleimide (NEM, 2.5 x 10(-5) M) inhibited the compound action potential of the phrenic nerve and increased the spontaneous release of transmitter from the nerve terminals, recorded as miniature endplate potentials. The first effect was the cause of a blockade of the phrenic nerve diaphragm preparation, during indirect stimulation. The left phrenic nerve was more susceptible to inhibition than the right. An increase of the threshold was observed during the progression of the inhibition. The inhibition was not use-dependent and there was no synergistic interaction with the local anaesthetic drug, tetracaine. The inhibition was partly antagonized by di-thio-threitol (3.0 x 10(-3) M). The increase of spontaneous release of transmitter was not accompanied by an increase of the stimulus-evoked release since the amplitude of the endplate potential was not increased and partial inhibition caused by d-tubocurarine or magnesium chloride was not antagonized. When the concentration of NEM was increased to 2.75 x 10(-4) M, the directly-elicited twitches were inhibited, and the baseline tension was increased. This increase of tension was slightly reduced in a preparation depolarized with potassium chloride; a small depolarization could partly explain this effect. It was not reduced by dantrolene or in a calcium-free solution. The inhibition of the twitch and the increased baseline tension (probably a rigor) might be caused by a reduced sensitivity of the contractile proteins for calcium ions and an inhibition of the myosin ATPase activity, respectively. PMID- 2571101 TI - Pentylenetetrazol kindling in mice. AB - Kindling with pentylenetetrazol to produce minimal and maximal convulsions was investigated in CF-1 mice. Like electrical kindling, the kindling effect was directly proportional to the dose or the intensity of the kindling stimulus. Similarly, the kindling effect was persistent, as was emphasized by the ability to kindle with an interdose interval of 3 days and by the convulsions produced by a challenge with pentylenetetrazol 30 days after withdrawal from the kindling treatment. The changes in excitability, associated with the kindling state, appeared to be relatively selective for pentylenetetrazol, because no changes in thresholds to either electroshock or administration of picrotoxin or N-methyl-DL aspartate correlated temporally with the persistence of kindling. The influence of two anticonvulsant drugs, ethosuximide and cannabidiol, on kindling was also investigated. Both drugs blocked the development of kindling to pentylenetetrazol induced minimal convulsions. Of these drugs, only ethosuximide raised the minimal convulsive threshold to pentylenetetrazol. Against pentylenetetrazol-induced kindling to maximal convulsions, only cannabidiol blocked kindling and only cannabidiol raised the maximal seizure threshold for pentylenetetrazol. Although the drugs modified the kindling effect, the mechanism of the interaction is not clear. PMID- 2571102 TI - Comparison of changes in the EEG of freely moving rats induced by enciprazine, buspirone and diazepam. AB - The effect of enciprazine, buspirone and diazepam was investigated on the cortical electrical activity in freely-moving rats. Enciprazine (5 mg/kg, i.p.) and buspirone (5 mg/kg, i.p.) induced comparable changes, consisting in decreases of mean power values in delta and theta and increases in alpha and beta EEG frequency bands. Regarding only a particular area of the brain or particular frequency bands, these two compounds could not be clearly separated from each other. Changes in frequency bands induced by O-methoxy-phenyl-piperazine (5 mg/kg i.p.) (D 15157), the presumed main metabolite of enciprazine, were dose-related to that caused by the parent compound. The second metabolite (R,S)-1-4-(1-methoxy 4-hydroxy-phenyl)piperazin-1-yl-3-(3,4,5- trimethoxyphenoxy)propan-2-ol dihydrochloride (D 20092) (5 mg/kg i.p.) evoked only minimal changes in the different frequency bands of the rats. The power spectra did not significantly differ from those seen in animals treated with saline. The action of diazepam (2 mg/kg i.p.) was characterized by decreases in alpha and delta frequency bands, accompanied by marked increases in fast beta waves. The marked frequency shifts caused by buspirone and enciprazine could clearly be differentiated from the EEG changes evoked by the minor tranquilizer, diazepam. PMID- 2571103 TI - Attenuation of SCH 23390-induced alteration of striatal dopamine D1 receptor ontogeny by prolyl-leucyl-glycinamide in the rat. AB - Long-term postnatal treatment of rats with SCH 23390 is associated with a reduction in the development of dopamine D1 receptors in the striatum. Because the tripeptide, L-prolyl-L-leucyl-glycinamide (PLG) attenuates the neuroleptic induced increase in D2 receptors in the striatum in adult rats, this study was undertaken with the objective of determining whether PLG could modulate a developmental alteration in the D1 subtype of receptor. Rats were treated with the dopamine D1 receptor antagonist, SCH 23390 (R[+]-7-chloro-8-hydroxy-3-methyl 1-phenyl-2,3,4,5-tetrahydro-1-H-3- benzazepine) (0.30 mg/kg/d i.p.) for 32 successive days from birth, while D1 receptors in the striatum were assessed at 5 and 8 weeks from birth. Postnatal treatment with SCH 23390 reduced in vitro binding of [3H]SCH 23390 to homogenates in the striatum by 70% at 8 weeks. Scatchard analysis at 5 weeks determined that the Bmax for the binding of [3H]SCH 23390 was reduced by 78%, while the Kd was unaltered. When PLG (1.0 mg/kg/d.i.p.) was administered together with SCH 23390 for the initial 32 days from birth, the binding of [3H]SCH 23390 to homogenates of the striatum was unchanged from that of the control group at 8 weeks. Also, at 5 weeks the Bmax and Kd were unaltered from control in the group that was treated with both SCH 23390 and PLG. The binding of [3H]SCH 23390 was not altered from control in the group treated with PLG alone. Also, PLG given in vitro did not alter the binding of [3H]SCH 23390 to control homogenates of the striatum. These findings indicate that PLG is able to attenuate neuroleptic-induced alterations in dopamine D1 receptors in the striatum. PMID- 2571104 TI - Investigation into the age-dependence of release of serotonin and noradrenaline in the rat brain cortex and of autoreceptor-mediated modulation of release. AB - Slices of cerebral cortex from young adult and aged rats, preincubated with [3H]serotonin or [3H]noradrenaline were superfused and the electrically (3 Hz) evoked overflow of tritium and its modulation by the respective agonists and antagonists were studied. The electrically evoked overflow in slices preincubated with [3H]serotonin from 2.5-year-old rats was smaller (by about 35%) than in slices from 3-month-old animals. No difference was found for the inhibition by serotonin and the facilitation by metitepine of the evoked overflow and for the metitepine-induced shift of the concentration-response curve for serotonin to the right. There was no difference between slices preincubated with [3H]noradrenaline from 3 months, 1, 2 and 2.5-year-old rats, with respect to the electrically evoked overflow, its inhibition and facilitation by noradrenaline and phentolamine, respectively, and the phentolamine-induced shift of the concentration-response curve for noradrenaline to the right. The present results suggest that the release of serotonin is reduced in the cerebral cortex of aged rats whereas the release of noradrenaline and the autoreceptor-mediated modulation of the release of either monoamine is not altered. PMID- 2571105 TI - Effect of cysteamine on levels of somatostatin-like immunoreactivity and catecholamines and on electroencephalogram in the rat brain. AB - Cysteamine (CYS) is known to be a quite specific depletor of somatostatin in the rat brain. In the present study we investigated the effect of CYS (100 mg/kg, 300 mg/kg, subcutaneously) on levels of somatostatin-like immunoreactivity (SLI) in the brain and cerebrospinal fluid, on catecholamines in the cortex, and on spectral cortical electroencephalogram (EEG) of rat. SLI was decreased in both the cortex and the striatum (p less than 0.05) of CYS-treated rats, but no change was seen in SLI of CSF. Cortical levels of dopamine, noradrenaline and homovanillic acid were decreased (p less than 0.05) following administration of either dose of CYS. In EEG, during mobility both the frontal and occipital peak (Fp) and mean (Fm) frequencies were slowed (p less than 0.05). Frontally, the amplitude of the frequency bands 1.5-3Hz and 3-5Hz was increased (p less than 0.05). During immobility the Fp and Fm were also slowed. In frequency bands of 3 5Hz, 5-10Hz and 10-20Hz the amplitude was decreased (p less than 0.05), indicating that, in addition to theta frequency, the low voltage fast activity is also affected by CYS. According to our results, both the cortical intrinsic neurons containing somatostatin and also the ascending catecholaminergic systems are affected after the single administration of CYS concomittantly with, but not necessarily related to, changes in different frequency bands in EEG. PMID- 2571106 TI - Effect of cysteamine on somatostatin-like immunoreactivity in the amygdala kindled rat brain. AB - Previous studies have shown that a somatostatin-depleting drug, cysteamine (CYS), suppresses kindled seizures. However, no data is available concerning the levels of somatostatin-like immunoreactivity (SLI) in the kindled rat brain after CYS administration. In the present study, we used radioimmunoassay to measure SLI in the frontal cortex, amygdala + piriform cortex, hippocampus, striatum and hypothalamus: 1) in control rats, 2) in amygdala-kindled rats decapitated 14 days after the last stimulus, and 3) in amygdala-kindled rats decapitated 14 days after the last stimulus but treated either 11 days or 4) 4 hours before decapitation with CYS (100 mg/kg, subcutaneously). The results showed that, compared to controls, in kindled rats SLI was elevated both in the ipsi lateral (28%, p = 0.0372) and contralateral (17%, p = 0.0078) frontal cortex. Compared to kindled rats, CYS given 4 hours before decapitation decreased SLI in the frontal cortex (to 71%, p = 0.0066) and hippocampus (to 72%, p = 0.0027), but compared to the controls, only in the hippocampus. In rats given CYS 11 days before decapitation, SLI did not differ from either the controls or from the kindled rats. In conclusion, the somatostatinergic system is affected in amygdala kindling; but the relationship of anatomical localization and the magnitude of CYS-induced decrease of SLI to elevated seizure threshold needs to be studied further. PMID- 2571107 TI - Hypothermia elicited by some prodynorphin-derived peptides: opioid and non-opioid actions. AB - Prodynorphin-derived peptides were tested for their effects on body temperature after intracerebroventricular administration to unrestrained male rats. Dynorphin A (Dyn A) (5 and 10 nmol) and Dynorphin A-(1-32) (Dyn A-(1-32) (2.5 and 5 nmol) lowered body temperature with a maximum approximately 30 min after administration. Dyn B (up to 50 nmol) did not induce hypothermia. Lower doses of all peptides did not alter body temperature. The hypothermic effect was significantly, but not completely prevented by MR1452 (30 nmol), a preferential antagonist of the kappa receptor, administered intracerebroventricularly. Naloxone, a mu receptor antagonist, naltrexone, its long acting analog up to doses of 100 nmol, as well as MR1453, the (+)-enantiomer of kappa antagonist MR1452 with no opioid binding properties, did not prevent the hypothermic effect. Moreover, episodic barrel rolling and bizarre postures elicited by Dyn A and Dyn A-(1-32) were reduced in rats pretreated i.c.v. with MR1452 (30 nmol), but not with naloxone (up to 100 nmol). Interestingly, des-Tyr-Dynorphin A (Dyn A-(2 17)), a fragment with virtually no opioid binding potential, was 4 times less potent that Dyn A in inducing hypothermia. These findings are consistent with the hypothesis that prodynorphin-derived peptides effects are not exclusively opioids in nature. PMID- 2571108 TI - [Endocrine neoplasms of the pancreas. Diagnostic aspects and treatment results]. PMID- 2571109 TI - L-homocysteic acid--a possible bipolar cell transmitter in the rabbit retina. AB - The identity of the retinal bipolar cell transmitter(s) is unknown although there is much indirect evidence that suggests it may be glutamate or a related compound. Some bipolar cells synapse onto cholinergic amacrine cells and in the rabbit retina acetylcholine (ACh) release is increased by light flashes and by the excitatory amino acids glutamate, aspartate and homocysteic acid (HCA). In the retina, the amino acid agonist N-methyl-D-aspartate (NMDA) is unusual in that it sometimes acts as an antagonist, and in the present experiments it blocked the light-evoked release of ACh by acting as an antagonist of the bipolar cell transmitter. However, NMDA did not block the actions of glutamate or aspartate on amacrine cell ACh release, a result that argues against either of these amino acids being the bipolar cell transmitter. On the other hand, the HCA evoked release of ACh was clearly antagonised by NMDA suggesting that HCA may be the bipolar cell transmitter released onto cholinergic amacrine cells. This suggestion is supported by the finding that the rabbit retina possesses HCA at a concentration of 0.8 nmol/g wet wt. PMID- 2571110 TI - Extracellular taurine increase in rat hippocampus evoked by specific glutamate receptor activation is related to the excitatory potency of glutamate agonists. AB - Taurine increases in brain extracellular space due to glutamate agonists were studied in vivo in the rat hippocampus using a dialysis technique, both in the absence and in the presence of glutamate receptor antagonists. Extracellular taurine levels increased during perfusions of agonists, listed in descending order of potency: kainate (KA), N-methyl-D-aspartate (NMDA), and quisqualate (QA). While taurine increases due to KA or QA perfusions were inhibited by 6,7 dinitro-quinoxaline-2,3-dione (DNQX), those induced by NMDA were abolished in the presence of 3-(carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP). These results indicate that increases in extracellular taurine levels evoked by NMDA, KA or QA in the rat hippocampus are caused by activation of their specific receptors. Field potentials, concomitantly recorded, were quickly abolished during NMDA or KA perfusions (0.1 mM), while QA (0.25 mM) induced the appearance of bicuculline-like evoked responses. Since taurine has been proposed as an osmoregulatory substance in the rat brain, and cell swelling is known to be an early component of glutamate agonists neurotoxicity, the increases in extracellular taurine reported here could be due to taurine released through an osmoregulatory process, counteracting the neurotoxic cellular oedema induced by glutamate agonists. PMID- 2571111 TI - Involvement of adenylate cyclase inhibition in dopamine autoreceptor regulation of tyrosine hydroxylase in rat nucleus accumbens. AB - In homogenates of rat nucleus accumbens, quinpirole, a dopamine (DA) D2 receptor agonist, inhibited the activation of tyrosine hydroxylase (TH) elicited by either forskolin, an activator of adenylate cyclase, or rolipram, a cyclic nucleotide phosphodiesterase inhibitor. The inhibition produced by 1 microM quinpirole was completely antagonized by the D2 blocker L-sulpiride (2 microM). Quinpirole failed to inhibit the stimulation of TH elicited by dibutyryl cyclic AMP (2 mM), which acts independently of adenylate cyclase. Quinpirole (10 microM) significantly inhibited the stimulation of adenylate cyclase activity elicited by 1 microM forskolin. These results indicate that mesolimbic DA autoreceptors can regulate TH activity by inhibiting a presynaptic adenylate cyclase system. PMID- 2571112 TI - Simultaneous recording of presynaptic spikes and excitatory postsynaptic potentials from monosynaptically connected hippocampal neurons. AB - A technique has been devised to activate single granule cells in the hippocampus, and to record simultaneously spikes from the particular granule cell and excitatory postsynaptic potentials from a monosynaptically connected CA3 neuron. The unitary excitatory postsynaptic potentials (EPSPs) sustained for long observation periods, and increased in size with increases in stimulus frequency and in external Ca2+ concentration. This technique may be useful for quantal analysis of transmission through the synapse between mossy fibers and CA3 neurons. PMID- 2571113 TI - One hundred years of retinoblastoma research. From the clinic to the gene and back again. AB - Retinoblastoma (RB) has provided the prototype for the study of hereditary predisposition to cancer. An intraocular tumour of young children, it has both hereditary and sporadic forms. The familial form of RB is inherited as an autosomal dominant with high penetrance. In a small group of patients the identification of a constitutional deletion on the long arm of chromosome 13 indicated the location of the critical gene in region 13q14 Close linkage between the hereditary, non-deletion form of RB and the esterase-D gene, which is also located in 13q14, demonstrated that all hereditary forms of RB are due to defects in a gene at this locus. Analysis of the development of homozygosity for region 13q14 in sporadic tumours subsequently confirmed that probably all RB tumors are due to mutations in a single gene, RB1. Using molecular biological techniques a candidate gene has been isolated which maps to region 13q14 and which shows structural re-arrangements within tumour cells. Predisposing, hereditary mutations have also been shown to involve this gene thereby providing strong evidence for its authenticity. The isolation of DNA sequences from within this gene, which identify restriction fragment length polymorphisms, means that it is now possible to use them to carry out pre-natal diagnosis and identify individuals at high risk to tumour development. PMID- 2571114 TI - Is it worthwhile to add dipivefrin HCl 0.1% to topical beta 1-, beta 2-blocker therapy? AB - In a prospective study, the addition of dipivefrin hydrochloride 0.1% twice daily to one eye of 32 patients with early primary open-angle glaucoma or ocular hypertension, maintained on a bilateral beta 1-, beta 2-blocker twice daily, resulted in a significant decrease of mean intraocular pressure (IOP) from 22.7 +/- 3.9 to 20.2 +/- 3.4 mmHg at 1 week (P = 0.0001) and to 21.0 +/- 3.8 mmHg at 12 weeks (P less than 0.02) in the dipivefrin-treated eyes. On the other hand, no significant change was noted in the fellow eyes (from 21.7 +/- 4.1 to 21.6 +/- 4.0 mmHg at 1 week and to 21.3 +/- 4.2 mmHg at 12 weeks). The addition of dipivefrin resulted in an IOP reduction of 2 mmHg or more in 50% and 3 mmHg or more in 19% of the eyes throughout the 12-week therapy. The result of the current study provides a realistic guideline as to what to expect from the common practice of adding dipivefrin hydrochloride to a beta 1-, beta 2-blocker regimen. PMID- 2571115 TI - Co-amplification of multiple regions of the HIV-1 genome by the polymerase chain reaction: potential use in multiple diagnosis. AB - We have used the polymerase chain reaction (PCR) to detect by co-amplification, multiple regions of the HIV-1 genome in infected cells. Genomic RNA and DNA from productively infected H9 cells were independently extracted and amplified in reactions with and without reverse transcriptase respectively using primer pairs to the gag, env, tat and nef regions of the viral genome in the same reaction mixture. PCR-products were analysed by liquid hybridization with end labelled oligonucleotide probes followed by gel-electrophoresis (oligomer hybridization). The primer pairs were capable of detecting as few as 10 copies of RNA and 10-20 copies of integrated proviral DNA. The ability to co-amplify multiple target regions in the same incubation mixture provides a method for detecting and confirming the presence of HIV-1 in samples for which limited nucleic acid is available. In addition, in reconstitution experiments, the same method was used to detect HIV-1 and HTLV-I simultaneously with comparable sensitivity (20-40 gene copies each). This offers the possibility of simultaneous diagnosis of multiple viral infections, such as those that occur in AIDS, on the same sample preparation. PMID- 2571116 TI - Reflex changes induced by clonidine in spinal cord injured patients. AB - In a single blind study of 6 spinal cord injured (SCI) men, the effects of clonidine, a selective alpha-2 adrenergic agonist, on spasticity were compared to diazepam and placebo. Since a potential side-effect of clonidine is postural hypotension, a combination of clonidine and desipramine was also tested. Vibration of the leg will inhibit the H reflex in a normal subject; whereas, this inhibition is markedly reduced in SCI patients with spasticity. A vibratory inhibition index (VII) was derived for each treatment. The pre-treatment VII was 92.08 +/- 3.15%; for SCI subjects, compared to 46.5 +/- 7.7% for 6 normal subjects. Clonidine significantly reduced the mean index of SCI patients to 59.42 +/- 3.91% (p less than 0.001). The VII for placebo, diazepam and the clonidine desipramine combination were not statistically different than the pre-treatment values in SCI patients. In conclusion, clonidine has an anti-spasticity effect in SCI patients, both subjectively, and objectively, in terms of vibratory inhibition of the H reflex. PMID- 2571118 TI - Release of endogenous excitatory amino acids and proteins by the mast cell degranulating peptide in the hippocampus. PMID- 2571119 TI - Palytoxin increases an outward K+ current in frog heart fibres. PMID- 2571117 TI - Cellular effects of beta-adrenergic and of cAMP stimulation on potassium transport in rat alveolar epithelium. AB - Alveolar fluid absorption is greatly enhanced by cAMP and by beta-adrenergic agonists via an increase in Na+ transport. Little is known about K+ homeostasis under these circumstances. We studied K+ transport across alveolar epithelium in isolated perfused rat lungs stimulated either by dibutyryl-cAMP or isoproterenol. K+ fluxes and the apparent permeability of 86Rb across the epithelium (alveoli to plasma) were interpreted according to a model involving two types of cells, B and L, distinguished by the location of Na+-K+-ATPases (basal and luminal). Water is considered to be absorbed by B cells in a solute-coupled process energized by a basolateral Na+-K+-ATPase that is stimulated by isoproterenol and cAMP. K+ transport out of the alveoli is due to the activity of a Na+-K+-ATPase located in the apical membrane of L cells. In the present study net transport rate of K+ was -0.5 +/- 0.15 nmol/s, n = 20 (out of alveoli) in control conditions. When the epithelium was stimulated by dibutyryl-cAMP (10(-4) mol/l) net absorption of K+ reversed to net 'secretion' into alveoli (3.2 +/- 0.31 nmol/s), fluid absorption was not stimulated. K+ 'secretion' was abolished by apical Ba2+, indicating it was due to opening of apical K+ channels. Basolateral ouabain reversed net K+ 'secretion' to net absorption indicating that K+ entry into alveoli was dependent on activity of B cell basolateral Na+-K+-ATPase (masking simultaneous K+ removal by apical L cell Na+-K+-pump). When larger concentrations of dibutyryl-cAMP (10( 3) mol/l) or when isoproterenol were used to stimulate the epithelium there was a tripling of fluid absorption.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571120 TI - Nucleotide sequence of PvuII polymorphic site at the human lipoprotein lipase gene locus. PMID- 2571121 TI - An X-linked DraI RFLP recognized by cpX23 [DXS132]. PMID- 2571122 TI - An EcoRV RFLP detected by probe pX59 [DXS77]. PMID- 2571123 TI - EcoRI polymorphism of the human immunoglobulin C epsilon 1 gene (IGHE1). PMID- 2571124 TI - An RFLP associated with insulinoma amyloid polypeptide locus (IAPP). PMID- 2571125 TI - A BamH1 RFLP of the C8B gene. PMID- 2571126 TI - Xmn-1 and Bg1-II RFLPs at the human hepatic lipase (HL) gene locus. PMID- 2571127 TI - Alterations in the pre-mRNA topology of the bovine growth hormone polyadenylation region decrease poly(A) site efficiency. AB - RNase mapping experiments show that the bovine growth hormone (bGH) poly(A) region forms an extensive hairpin loop. Mutants were prepared to change poly(A) region pre-mRNA structure and cleavage site efficiency without altering necessary sequences. An inverted repeat which includes the poly(A) cleavage site was created by insertion of a linker upstream of the poly(A) region to compete with any wild-type secondary structure. RNA mapping analyses show alterations in the nuclease accessibility of this mutant at the natural site of cleavage. This mutant shows a 75% drop in relative reporter gene expression at the steady-state protein and RNA levels. When the linker is inserted as a direct repeat, expression is equivalent to wildtype levels. To show that transcription was not terminated by the inverted repeat, the SV40 late poly(A) region was inserted downstream. These mutants show restored expression and processing at the downstream site. Our experiments reveal that the conformation of the poly(A) site pre-mRNA is important in mediating efficient cleavage-polyadenylation. PMID- 2571129 TI - DNA amplification for detection of the XV-2c polymorphism linked to cystic fibrosis. PMID- 2571130 TI - A new anonymous marker on chromosome 7, D7S420, identifies a PvuII RFLP. PMID- 2571131 TI - A new anonymous marker on chromosome 11, D11S347, detects two TaqI RFLPs. PMID- 2571128 TI - Structure and methylation of the human calcitonin/alpha-CGRP gene. AB - We report a detailed analysis of the human calcitonin/alpha-CGRP gene locus. About 39kb of DNA containing the gene has been mapped and a common Pvu II RFLP identified downstream of the gene. DNA sequence analysis revealed an extensive CpG island containing several rare restriction enzyme sites at the 5' end of the gene. The structure of this island is unusual in that it contains two distinct CpG-rich regions, one located around exon 1 and the other about 1.5kb further upstream. Msp I sites within both CpG-rich regions were found to be unmethylated, regardless of whether the calcitonin/alpha-CGRP gene was being expressed. However, a correlation was found between demethylation of Msp I sites in intron 2, downstream of the CpG island, and calcitonin/alpha-CGRP gene expression. DNA sequence analysis also revealed the presence of several binding sites for constitutive and regulatory transcription factors in the promoter of the gene. These results suggest that both unmethylated CpG islands and specific demethylation of internal sequences may play a role in the activation of calcitonin/alpha-CGRP gene transcription. PMID- 2571132 TI - An anonymous single copy probe, D3S153, detects an SstI RFLP. PMID- 2571133 TI - A new marker on chromosome 4q, D4S138, detects two RFLPs. PMID- 2571134 TI - A new polymorphic probe on chromosome 3p: lambda LIB34-60 [D3S154]. PMID- 2571135 TI - A new polymorphic probe on chromosome 3p: lambda LIB49-60 [D3S155]. PMID- 2571136 TI - A polymorphic DNA probe from chromosome 19 (19cen-q11). PMID- 2571137 TI - Drug withdrawal. PMID- 2571139 TI - Neuropeptide tyrosine (NPY)-induced potentiation of the pressor activity of catecholamines in conscious rats. AB - IV bolus administration of 2.5-50 micrograms NPY (0.6-12.5 nmol) to conscious rats produced a dose- and time-dependent increase in systolic and diastolic blood pressure. Following priming with 2.5 micrograms NPY, or larger doses, the subsequent administrations of noradrenaline produced pressor responses that were potentiated both in magnitude and duration. The NPY-induced potentiation of the pressor response to noradrenaline was dose-dependent and extended to the pressor action of adrenaline and angiotensin II but not to the hypotensions produced by bradykinin or isoproterenol. The potentiation was not related to the fact that multiple doses of catecholamines were repeated. Reserpine did not substantially modify the NPY-induced potentiation of the pressor activity of the catecholamines. Chemical sympathectomy following 6-hydroxydopamine caused a marked supersensitivity to the catecholamines and NPY but obliterated the NPY induced potentiation of the pressor effect of adrenaline. Nifedipine reduced the pressor action of the catecholamines and NPY but did not attenuate the NPY induced potentiation of the pressor action of catecholamines. It is concluded that the acute pressor effect of NPY and of the potentiation of the catecholamine pressor effects involve different mechanisms. PMID- 2571140 TI - Amiloride enhances atrial natriuretic factor stimulation of cGMP accumulation in rat glomeruli. AB - The effects of amiloride on ANF binding and ANF stimulation of cGMP were evaluated in rat glomeruli. Amiloride increased ANF binding to whole glomeruli and to glomerular membrane preparations. In contrast, amiloride enhanced ANF stimulated cGMP accumulation only at 37 degrees C in whole glomeruli, but not at 4 degrees C in whole glomeruli or at 37 degrees C in membrane preparations. These data suggest that under physiological conditions amiloride augments ANF stimulated intracellular cGMP accumulation. The discrepancies between amiloride augmentation of ANF binding and failure to increase ANF-stimulated cGMP accumulation may result from ANF receptor heterogeneity. This is the first report of an amiloride augmentation of ANF-stimulated cGMP accumulation in renal tissue. PMID- 2571138 TI - Differential inhibitory effects of MIF-1, Tyr-MIF-1, naloxone and beta funaltrexamine on body rotation-induced analgesia in the meadow vole, Microtus pennsylvanicus. AB - The effects of body rotation in a horizontal plane and various opiate antagonists on the nociceptive responses of a day-active microtine rodent, the meadow vole, Microtus pennsylvanicus, were examined. Intermittent rotation (70 rpm, schedule of 30 sec on, 30 sec off) for 30 min induced significant analgesic responses in the voles for 15 min after rotation. These increases in thermal response latency were blocked by intraperitoneal pretreatment with either naloxone or the irreversible mu opiate receptor antagonist beta-funaltrexamine (beta-FNA; 10 mg/kg; 24 hr pretreatment). This antagonistic effect of beta-FNA indicates mu opioid involvement in the mediation of rotation-induced analgesia. The antiopiate peptides MIF-1 (Pro-Leu-Gly-NH2) and Tyr-MIF-1 also significantly reduced, though did not completely block, body rotation-induced opiate analgesia. This suggests that Tyr-MIF-1 and MIF-1 have significant antagonistic effects on mu opioid systems that are involved in the mediation of stress (rotation)-induced analgesia. PMID- 2571141 TI - [Evaluation of the usefulness of forced diuresis in patients with calcium cyclobarbital or reladorm poisoning]. AB - An attempt was made at evaluating the usefulness of forced diuresis for the treatment of Phanodorm poisoning. Fifty two patients were examined, 40 of them with Reladorm (Polish trade name. Reladorm contains 100 mg of cyclobarbital and 10 mg of diazepam) poisoning and 12 with Cyclobarbitalum Calcium (Phanodorm) poisoning. The test group was selected out of those poisoned with barbiturates so that a homogeneous toxic agent would be involved and the impact that other toxins may have on the rate of metabolism and excretion eliminated. The elimination of drug from blood was compared with the elements determining ED intensity, such as i.v. fluid administration, primary infiltrate (clearance of endogenous creatinine), reabsorption in renal tubules and diuresis rate. No correlation was found which is an evidence that ED proves to be useless in the treatment of Phanodorm poisoning. The author suggests that in this type of poisoning supportive treatment should be undertaken according to daily fluid demand in relation to such factors as sex, age, body temperature, health conditions and contraindications due to the patient's general condition. PMID- 2571142 TI - Allergic blepharoconjunctivitis. Avoiding misdiagnosis and mismanagement. PMID- 2571143 TI - Therapeutic uses of beta-adrenoceptor blocking drugs in the central nervous system in man. PMID- 2571144 TI - Influence of beta-agonist on plasma concentrations of growth hormone in broiler chickens on a low plane of nutrition. AB - The effect of the chronic administration of a beta-agonist (L-640,033; donated by Merck, Sharp and Dohme, Research Laboratories, Rahway, NJ) on both the plasma concentration of growth hormone (GH) and the episodic pattern of GH secretion was examined. Administration of .25, 1.0, or 4.0 ppm beta-agonist in the diet for only 3 to 5 days did not affect the overall mean plasma concentration of GH. These treatments also did not influence the frequency of GH secretory pulses. However, the amplitude of the GH secretory pulses was reduced. In contrast, administration of 1.0 ppm beta-agonist for the 10 to 12-day period increased the mean plasma concentration of GH, the amplitude of the GH secretory pulses, the basal (between pulses) plasma concentration of GH, and the interpeak interval. No effect of in vivo GH secretion was found with .25 ppm beta-agonist treatment for 10 to 12 days. Chicks receiving 4.0 ppm beta-agonist for 10 to 12 days had GH secretory pulses with increased amplitude, but no other differences in GH secretory characteristics were observed. PMID- 2571145 TI - Peripheral and central control of food intake. AB - To either increase or decrease body weight of poultry, an understanding of food intake regulation is essential. Although it is advantageous to increase food intake in market birds, it is desirable to decrease intake in breeders. Recent studies have shown that the digestive tract, liver, and brain are all involved in food intake regulation. In this review, the role of various neurotransmitters and metabolic substrates in food intake regulation, both within the central nervous system as well as in the periphery, is discussed. In addition, how the strain of the bird or the physiological state of the animal influences the response to various compounds is discussed. PMID- 2571146 TI - Maternal tobacco smoking and changes in amino acid uptake by human placental villi: induction of uptake systems, gammaglutamyltranspeptidase and membrane fluidity. AB - Maternal smoking depressed the active uptake of amino acids by human placentae and lowered their levels in the placenta and umbilical vein. During starvation of cells for amino acids, more amino acid carriers are induced and incorporated into the plasma membrane. A question arises whether there could be similar changes due to maternal smoking in the placental amino acid uptake carrier systems. Therefore, the characteristics of (a) the uptake of 2-amino[I-14C]-isobutyric acid (AIB) by isolated placental villi, (b) gammaglutamyltranspeptidase (GGTP), a critical enzyme of the gammaglutamyl cycle (GGC) for the uptake of amino acids in human placenta, and (c) lipid structural parameters (reciprocal of fluidity), by steady state fluorescence polarization of plasma membrane vesicles of microvilli (MV) and microsomal membranes (MM) of umbilical and chorionic plate arteries of placentae of smoking and non-smoking mothers were investigated. The above investigations gave the following results: (a) Washed placental villi of smokers exhibited higher capacity for AIB uptake than those of non-smokers. The higher uptake capacity was mainly due to increase in Vmax for AIB uptake in smokers. Km increased for placental AIB uptake in smokers. (b) Maternal smoking lowered GGTP activity of MV by decreasing its Vmax. Therefore, maternal smoking decreases the formation of gammaglutamyl-amino acid (GGAA) on the surface of trophoblast which are absorbed by the trophoblast. The degree of absorption of GGAA is considered as an inverse environmental signal for the cell to regulate amino acid transport systems. Maternal smoking seems to decrease the formation and absorption of GGAA and thereby induce the formation of new carriers for AIB uptake. (c) Maternal smoking increased the values for lipid structural order parameters and microviscosity of MV and induced tolerance against fluidization by ethyl alcohol in MM of umbilical and chorionic arteries. The alterations could increase Km for AIB uptake system and decrease the sensitivity of umbilical and chorionic arteries to vasoconstrictive substances like 5-hydroxytryptamine and catecholamine which are released by nicotine. All these changes tend to overcome the deficits produced in placental amino acid transport and satisfy the demands of the growing fetus for amino acids. PMID- 2571147 TI - A new approach using DNA fingerprinting for the determination of androgenesis as a cause of hydatidiform mole. AB - A new method of DNA analysis has been used for the determination of androgenesis as a cause of complete hydatidiform mole. This method, using a minisatellite core probe, requires only a small amount of DNA and detects the restriction fragment length polymorphisms (RFLPs) due to allelic differences in the number of tandem repeats containing the core sequence. Southern blot hybridization showed an individual-specific DNA fingerprint, and each polymorphic band in molar tissues could be identified as being of paternal, but not maternal, origin. Some polymorphic bands of paternal DNA were not observed in molar tissues, indicating that endoreduplication of a normal haploid sperm or fertilization by dispermy to an anuclear oocyte with no effective genome could be the cause of complete hydatidiform mole. This method is sufficiently reliable and rapid that differential diagnosis could be made between complete hydatidiform mole, partial mole and hydropic change. PMID- 2571148 TI - Expression of major histocompatibility complex class I antigens in rat muscle cultures: the possible developmental role in myogenesis. AB - The expression of major histocompatibility complex class I antigens was demonstrated on aneurally cultured rat muscle cells. Myoblasts showed constitutive expression of class I antigens on their cell surfaces. The presence of the antigens was transitory, disappearing as myoblasts fused and differentiated into multinucleate myotubes. Furthermore, antibody against rat class I antigens showed an inhibitory effect on the generation of myotubes during muscle development. Although mature myotubes did not show any detectable levels of class I antigens on their cell surface, soluble factors from concanavalin A activated spleen cells or interferon gamma could induce the expression of class I antigens on muscle fibers. These results suggest that the expression of class I antigens on muscle cells is not only immunologically modulated but also developmentally regulated and that the antigens may play a role in cell recognition and interactions during the fusion process of myogenesis. PMID- 2571149 TI - Duplication of large genomic regions during the evolution of vertebrate homeobox genes. AB - The phylogenetic relationships of 21 murine Antp-class (Drosophila mutation Antennapedia-type class) homeobox genes have been analyzed, and several groups of related genes have been identified. The murine Antp-class homeobox genes are localized within four gene clusters. The similar structural organization of the four gene clusters strongly suggests that genes within a group of related Antp class homeobox genes are derived from duplications of large genomic regions. After the duplication, the gross structures of the homeobox gene clusters have been maintained over a long period of evolutionary time, indicating that the specific organization of genes within a cluster may be of functional importance. PMID- 2571150 TI - Primarily chronic progressive and relapsing/remitting multiple sclerosis: two immunogenetically distinct disease entities. AB - HLA class II gene polymorphism was investigated in 100 patients with clinically definite multiple sclerosis (MS) by restriction fragment length polymorphism analysis of Taq I-digested DNA using DRB, DQA, and DQB cDNA probes. Twenty-six patients had primarily chronic progressive MS and 74 had relapsing/remitting MS. The latter group included patients with a secondary progressive evolution of symptoms. Both clinical forms of MS were found to be associated with the DRw15,DQw6 haplotype. In addition, primarily chronic progressive MS was positively associated with the DQB1 restriction fragment pattern seen in DR4,DQw8, DR7,DQw9, and DRw8, DQw4 haplotypes, as well as negatively associated with the Taq I DQB1 allelic pattern corresponding to the serological specificity DQw7. Relapsing/remitting MS was positively associated with the DQB1 allelic pattern observed in the DRw17,DQw2 haplotype. These three DQB1 alleles are in strong negative linkage disequilibria with DRw15. The two susceptibility markers of each clinical form of MS act additively in determining the genetic susceptibility, as the relative risks for individuals carrying both markers roughly equal the sum of respective risks. Different alleles of the DQB1 locus defined by restriction fragment length polymorphisms contribute to susceptibility and resistance to primarily chronic progressive MS as well as to susceptibility to relapsing/remitting MS. The observed immunogenetic heterogeneity between the different clinical forms of MS favors the hypothesis that primarily chronic progressive MS and relapsing/remitting MS are two distinct disease entities. PMID- 2571152 TI - Retroviral transfer of a human tyrosine hydroxylase cDNA in various cell lines: regulated release of dopamine in mouse anterior pituitary AtT-20 cells. AB - Little is known about the molecular events mediating neurotransmitter release, a crucial step in synaptic transmission. In this paper, the biosynthesis and release of L-beta-3,4-dihydroxyphenylalanine (L-DOPA) and dopamine were analyzed in three heterologous cell lines after retroviral-mediated gene transfer of tyrosine hydroxylase (EC 1.14.16.2), the rate-limiting enzyme in catecholamine synthesis. A recombinant retrovirus encoding human tyrosine hydroxylase type I as well as neomycin-resistance gene was used to infect a fibroblast (NIH 3T3), a neuroblastoma (NS20 Y), and a neuroendocrine (AtT-20) cell line. After selection in the presence of neomycin and in tyrosine-free medium, high levels of exogenous tyrosine hydroxylase activity were detected in extracts of the three cell lines. High-performance liquid chromatography of cell extracts and culture supernatants confirmed that the three cell lines hydroxylated tyrosine to form L-DOPA and released this metabolite into the culture medium. Interestingly, the neuroendocrine cell line AtT-20 synthesized not only L-DOPA but also dopamine. Evoked secretion studies established that AtT-20 cells released the transmitter upon depolarization in a regulated, calcium-dependent way. We discuss the implication of this approach for the analyses of neurotransmitter release as well as in the context of degenerative disorders such as Parkinson disease. PMID- 2571151 TI - Loss of distinct regions on the short arm of chromosome 17 associated with tumorigenesis of human astrocytomas. AB - Astrocytomas, including glioblastoma multiforme, represent the most frequent and deadly primary neoplasms of the human nervous system. Despite a number of previous cytogenetic and oncogene studies primarily focusing on malignant astrocytomas, the primary mechanism of tumor initiation has remained obscure. The loss or inactivation of "tumor suppressor" genes are thought to play a fundamental role in the development of many human cancers. Thus, we have analyzed astrocytomas of various histological malignancy grades with polymorphic DNA markers to search for specific chromosomal deletions potentially pointing to loci containing tumor suppressor genes. Loss of constitutional heterozygosity indicating chromosomal loss or deletions was most frequently seen for markers on the short arm of chromosome 17 in 50% of the informative tumors (5 of 10 informative cases) and, to a lesser extent, for markers on chromosomes 1 and 10. Deletions on chromosome 17p were seen in both low-grade and high-grade malignant astrocytomas, suggesting that this chromosome may contain a tumor suppressor gene associated with the early events in tumorigenesis. The common region of deletions on the short arm of chromosome 17 is, therefore, clearly distinct from the gene causing von Recklinghausen neurofibromatosis (NF1), a tumor syndrome associated with glial tumors that maps to the long arm of chromosome 17. The search for progressively smaller deletions on chromosome 17p in astrocytomas may be the way to clone and characterize this locus, thus leading to insights into normal and abnormal growth and differentiation of glial cells. PMID- 2571153 TI - Fatty acid modifies Ca2+-dependent potassium channel activity in smooth muscle cells from the human aorta. AB - By using the patch-clamp technique the effect of 2-decenoic acid (DA) on Ca2+ activated potassium (K+) channels in the membrane of smooth muscle cells from the human aorta was studied. In the presence of 0.5 microM Ca2+ and 2 mM Mg2+ on the cytoplasmic side of the membrane, a more than tenfold elevation in the probability of the channels being open (po) was observed under the effect of DA. With divalent cation concentrations of less than 1 nM DA caused a more than twofold elevation in po. In the DA-treated membranes Mg2+ ions, which normally fail to activate the channels, brought about a nearly threefold increase in the channel activity when applied to the inner membrane surface. Channel sensitivity to the activating effect of cytoplasmic Ca2+ ions did not increase with the application of DA. Single-channel conductance was unchanged by DA exposure. We suggest that DA alters the Ca2+-binding mechanism of the channel, increasing its sensitivity to Mg2+ ions, presumably owing to membrane fluidization. PMID- 2571155 TI - The assembly of ionic currents in a thalamic neuron. II. The stability and state diagrams. AB - In the previous model of a thalamic neuron (R.M. Rose & J.L. Hindmarsh, Proc. R. Soc. Lond. B237, 267-288 (1989], which we referred to as the z-model, the burst response was terminated by the slow activation of a subthreshold outward current. In this paper we show that similar results can be obtained if the burst response is terminated by slow inactivation of the subthreshold inward current, Isa. We illustrate the use of this new model, which we refer to as the ha-model, by using it to explain the response of a thalamic neuron to a double ramp current. The main aim of the paper is to show how the stability and state diagrams introduced previously can be used to explain various types of firing pattern of thalamic and other neurons. We show that increasing the threshold for the fast action potentials leads to low threshold spikes of increased amplitude. Also, addition of a second subthreshold inward current adds a new stability region, which enables us to explain the origin of plateau potentials. In addition, various types of subthreshold oscillation are produced by relocating a previously stable equilibrium point in an unstable region. Finally, we predict a sequence of responses to current steps from different levels of background current that extends the burst, rest, tonic sequence of thalamic neurons. The stability and state diagrams therefore provide us with a useful way of explaining further properties of thalamic neurons and appear to have further applications to other mammalian neurons. PMID- 2571154 TI - The assembly of ionic currents in a thalamic neuron. I. The three-dimensional model. AB - We have previously discussed qualitative models for bursting and thalamic neurons that were obtained by modifying a simple two-dimensional model for repetitive firing. In this paper we report the results of making a similar sequence of modifications to a more elaborate six-dimensional model of repetitive firing which is based on the Hodgkin-Huxley equations. To do this we first reduce the six-dimensional model to a two-dimensional model that resembles our original two dimensional qualitative model. This is achieved by defining a new variable, which we call q. We then add a subthreshold inward current and a subthreshold outward current having a variable, z, that changes slowly. This gives a three-dimensional (v,q,z) model of the Hodgkin-Huxley type, which we refer to as the z-model. Depending on the choice of parameter values this model resembles our previous models of bursting and thalamic neurons. At each stage in the development of these models we return to the corresponding seven-dimensional model to confirm that we can obtain similar solutions by using the complete system of equations. The analysis of the three-dimensional model involves a state diagram and a stability diagram. The state diagram shows the projection of the phase path from v,q,z space into the v,z plane, together with the projections of the curves z = 0 and v = q = 0. The stability of the points on the curve v = q = 0, which we call the v, q nullcurve, is determined by the stability diagram. Taken together the state and stability diagrams show how to assemble the ionic currents to produce a given firing pattern. PMID- 2571156 TI - The assembly of ionic currents in a thalamic neuron. III. The seven-dimensional model. AB - We replace our earlier three-dimensional ha-model of a thalamic neuron (Rose & Hindmarsh, Proc. R. Soc. Lond. B 237, 289-312 (1989b)) by a seven-dimensional ha model. The stability and state diagrams for this seven-dimensional model are shown to be similar to those of the three-dimensional system with which it is compared. Two examples that illustrate how the seven-dimensional model can be related to experimental recordings are then discussed in detail. In each case we show the state and stability diagrams and the time courses of the different ionic currents during the burst response. We discuss how the various components of the state diagram could be determined experimentally. These experiments are illustrated by using our model to simulate the results of actual experiments. Finally the state and stability diagrams are transferred to a current-voltage diagram. This shows the advantage of the state diagram compared with the current voltage diagram for representing the dynamics of the firing of a thalamic neuron. PMID- 2571157 TI - Osmosis in leaky pores: the role of pressure. AB - Osmosis in leaky pores is often considered to involve gradients of pressure that originate from concentration gradients at the pore wall. These postulated surface pressures are incompatible with the Boltzmann equation and result from an incorrect application of the Gibbs-Duhem equation to binary solutions near surfaces. Such pressures would give rise to tensions in the surface phase that are not observed experimentally. Application of the argument to the swelling of porous systems and gels shows that this phenomenon cannot involve internal pressures within the fluid phase. PMID- 2571158 TI - Osmotic flow equations for leaky porous membranes. AB - A basic set of equations describing the flows of volume (Jv) and solute (Js) across a leaky porous membrane, coupled to the differences of osmotic and hydrostatic pressures d pi and dP has been derived by using general frictional theory. Denoting the mean pore concentration of solute by c*s and the hydraulic and diffusive conductances by Lp and Ps/RT the equations take the form Jv = LpdP + sigma sLp d pi Js = c*s(1 - sigma f)Jv + Ps d pi/RT sigma s = theta (1 - DsVs/DwVw - Ds/Dos) sigma f = 1 - theta DsVs/DwVw - Ds/Dos in which Dw and Ds are the diffusion coefficients for water and solute in the pore and Dos that for free solution. The relation between the reflection coefficients sigma s and sigma f for osmosis and ultrafiltration is then given by sigma s = sigma f - (1- theta)(1 - Ds/Dos), where theta is the diffusive-driven:pressure-driven flow ratio. These equations follow from the fact that in leaky pores osmosis occurs by diffusion alone and that there cannot be any Onsager symmetry leading to sigma s = sigma f. Symmetry holds in the limits where either the pore is small, when sigma s = sigma f = 1, or where the pore is large when sigma s = sigma f = 0. PMID- 2571159 TI - Ionic and charge-displacement currents evoked by temperature jumps in Xenopus oocytes. AB - Membrane currents were recorded in voltage-clamped oocytes of Xenopus laevis. Currents were produced in response to temperature jumps imposed by a heating lamp. Responses were larger when the animal (pigmented) hemisphere of the oocyte was illuminated as compared to the vegetal hemisphere; they arose because of a thermal effect as they were attenuated by removal of infrared wavelengths. The temperature jump responses comprised two distinct components: (i) a slow maintained current, which inverted direction at a membrane potential of about -25 mV and, (ii) a fast transient current, which at all potentials examined (-160 to +30 mV), was inward at the onset of a light flash and outward at the offset. The slow component probably arises through temperature-dependent changes in the 'leakage' current of the oocyte, and measurements of reversal potentials in solutions of different ionic composition indicated that currents carried by Na+ and H+ ions contribute to the response. In contrast, the fast component was not altered by changes in composition of the bathing solution. This observation, together with the finding that the charge movements associated with the on and off transients were of similar magnitude, suggest that the fast current may arise because of the displacement of charges across the plasma membrane. PMID- 2571160 TI - Effects of dopaminergic and GABAergic antagonists on methamphetamine-induced locomotor activity in mice selectively bred for their differences in locomotor activity. PMID- 2571161 TI - Interaction between several S1A and alpha 1 agonists on rabbit aortic rings. PMID- 2571162 TI - The effects of the alpha 1-adrenergic antagonists terazosin and phentolamine on prostatic adenoma and urethra in vitro. PMID- 2571163 TI - Alteration in the pharmacologic activity of alpha 1 adrenergic antagonists by alpha-1-acid glycoprotein. PMID- 2571164 TI - Beta-adrenergic binding sites on alpha 1-acid glycoprotein and mononuclear leukocytes. PMID- 2571165 TI - Tertatolol high affinity binding to alpha 1-acid glycoprotein: stereospecificity and consequences on plasma binding in disease states. PMID- 2571166 TI - Prospects for correction of thalassemia by genetic engineering. AB - The thalassemias are diverse genetic disorders characterized by abnormal synthesis rates of one or more proteins constituting hemoglobin (globin-chains). In the beta-thalassemias, genes encoding the beta-globin chain are intact but are abnormally transcribed or, less often, translated. In the alpha-thalassemias, genes encoding the alpha-globin chain are often deleted; abnormal transcription can also occur. The human beta- and alpha-globin genes were molecularly cloned. This review considers attempts to introduce these genes in mammalian cells by physical techniques such as chromosome transfer, transfection, fusion, micro injection, electroporation and homologous recombination or by using DNA or RNA viruses, such as retroviruses. Currently, inefficient gene expression in host cells and the need for precise cognate regulation of globin gene expression are the major limitations to applying genetic engineering to thalassemia. PMID- 2571167 TI - Stimulation and inhibition of food intake by the selective dopamine D2 agonist, N 0437: a meal pattern analysis. AB - Feeding and drinking responses were recorded in home-caged rats over 24-hour periods, and the data were analysed in terms of meal patterns. The highly selective dopamine D2 receptor agonist, N-0437, was injected IP at the start of the night phase. At the smallest dose (0.3 mg/kg), N-0437 increased food intake as a result of increases in meal size and duration. The rate of feeding during meals was not increased. This effect lasted 1-2 hours in the night phase. At higher doses (1.0 and 3.0 mg/kg), N-0437 inhibited food intake for 3-6 hours. The inhibition was due to a substantial reduction in meal size, with no change in meal frequency. Rate of feeding during meals tended to be reduced. A biphasic dose-effect relationship was not apparent in the drinking data, and there was not a significant effect of N-0437 on drinking responses. The meal pattern analysis results indicate the stimulation of D2 receptors produces increases and decreases in food intake and meal size. These results are discussed in terms of dose dependent stimulation of presynaptic (autoreceptor) and postsynaptic D2 receptors, and the relationship to satiety within meals. PMID- 2571168 TI - Spinal vs. supraspinal sites of action of the alpha 2-adrenergic agonists clonidine and ST-91 on the acoustic startle reflex. AB - Previous work has shown that stimulation of alpha 2-adrenergic receptors depresses the startle responses in rats. The present study suggests that this depressant effect involves supraspinal rather than spinal alpha 2-adrenergic receptors because intraventricular but not intrathecal infusion of the hydrophilic alpha 2-adrenergic agonist ST-91 depressed the acoustic startle reflex. To determine the point in the acoustic startle pathway where alpha 2 adrenergic receptor activation might ultimately alter neural transmission, startle responses were elicited electrically from different points along the acoustic startle pathway after systemic administration of clonidine. Clonidine depressed acoustically-elicited startle and startle elicited by electrical stimulation of the ventral cochlear nucleus to a comparable magnitude and over a similar time course. It also partially depressed startle elicited by electrical stimulation of the nucleus reticularis pontis caudalis (RPC). Taken together, these data suggest that alpha 2-adrenergic stimulation depresses startle by acting on supraspinal receptors, but that this effect is ultimately expressed, at least in part, by actions at both spinal and brainstem levels of the acoustic startle response pathway. The results are compared to other drugs known to affect the startle reflex. PMID- 2571169 TI - MIF-1 and Tyr-MIF-1 fail to alter benzodiazepine-induced hypothermia. AB - Tyr-MIF-1 (Tyr-Pro-Leu-Gly-NH2) and MIF-1 (Pro-Leu-Gly-NH2) can increase GABA stimulated benzodiazepine binding in brain tissue can block the hypothermia induced by several other compounds. Since benzodiazepines can also cause hypothermia, colonic temperatures were measured in mice after administration of chlordiazepoxide (CDP) and these two brain peptides. In several experiments involving different doses and times of administration of CDP, MIF-1, and Tyr-MIF 1, there were no significant effects of the peptides in altering the reliable decrease in colonic temperature induced by the benzodiazepine. The results indicate that the interaction of Tyr-MIF-1 and MIF-1 with benzodiazepines does not involve thermoregulation. PMID- 2571170 TI - Anticonvulsant and other effects of diazepam grow with time after a single treatment. AB - The hypothesis was tested that some of the effects in rats of the prototypical benzodiazepine, diazepam, would grow (i.e., sensitize) with the passage of time after acute administration as we had previously observed following stimulants, antidepressants, neuroleptics and other compounds. Our principal findings indicate that: 1) A single pretreatment with 0.5 mg/kg of diazepam significantly enhances the anticonvulsant effect of this same dose administered again two weeks later. 2) One injection of 2.5 mg/kg of diazepam significantly sensitizes the catalepsy and ptosis observed following the administration of haloperidol two weeks but not two hours later. These data provide the first evidence for time dependent sensitization after benzodiazepines and perhaps by implication, of GABA neurons. They may also suggest that acute stimulation of GABA neurons triggers the progressive development of a long-term, antidopaminergic influence. Finally, they raise the question of whether the progressive anxiolytic influence seen during the first week or so of benzodiazepine therapy depends on the passage of time rather than repeated drug treatment. PMID- 2571171 TI - Behavioural synergism between the dopamine agonists SKF 38393 and LY 171555 in dopamine-depleted mice: antagonism by sulpiride reveals only stimulant postsynaptic D-2 receptors. AB - The behavioural effects of the D-1 agonist SKF 38393 (0.56-45 mg/kg) and the D-2 agonist LY 171555 (0.05-6.25 mg/kg) were studied in mice rendered akinetic with a combination of reserpine and alpha-methyl-p-tyrosine (alpha-MPT). Under these conditions both agonists were behaviourally ineffective by themselves, but interacted synergistically to restore locomotor and orofacial movements. High levels of D-1 stimulation did not promote stereotypies at the expense of locomotion in dual reserpine- and alpha-MPT-treated mice, suggesting that the concomitant stimulation of D-1 receptors is essential both for the genesis and subsequent development of all components of D-2 behaviours. The motor stimulant effects of LY 171555 (0.05-1.25 mg/kg), administered in conjunction with a near maximal dose of SKF 38393 (15 mg/kg), were dose-dependently inhibited by sulpiride at all doses tested (1, 10 and 50 mg/kg). The augmentation of D-2 behaviours by low doses of sulpiride, noted in an earlier study, was not observed here. Results with this particular combination of drugs therefore do not support the concept of mixed excitatory and inhibitory postsynaptic D-2 receptors. The results are discussed in terms of different D-2 agonists interacting with discrete subpopulations of D-2 receptors at postsynaptic sites. PMID- 2571172 TI - Regional concentrations of transmitter amino acids after spinal cord ischaemia in the rabbit. AB - Concentrations of Asp, Glu, Gly, GABA and Gln were studied in the ventral and dorsal horns of the rabbit spinal cord after ligation of the abdominal aorta. The most significant changes observed after 10, 20 and 40 min ischaemia were an increase in the Asp and GABA concentration in the ventral horns and an increase in the Asp, Gly and GABA concentration in the dorsal horns. These changes correspond to shifts in the relevant reactions under conditions of the altered redox equilibrium in the tissue during ischaemia. Four days after 10 min ischaemia, amino acid concentrations in the spinal cord were at the control levels. Four days after 20 and 40 min ischaemia Asp, Gly and GABA concentrations were decreased in the ventral horns and Asp, Gly, GABA and Glu concentrations in the dorsal horns. The percentually greater decrease in the concentration in the ventral horns may be associated with the greater morphological damage to these structures. PMID- 2571173 TI - Discriminative stimulus effects of the alpha 2-adrenoceptor antagonist idazoxan. AB - Rats were trained to discriminate a dose of the alpha 2-adrenoceptor antagonist idazoxan (10 mg/kg IP) from saline. The discriminative stimulus produced by idazoxan was dose related and generalised to yohimbine. However, generalisation did not occur with a variety of compounds from other pharmacological categories including the alpha 1-adrenoceptor agonist cirazoline, the alpha 1-adrenoceptor antagonist prazosin, and the alpha 2-adrenoceptor agonist clonidine. The idazoxan stimulus was not antagonised by either prazosin or clonidine, although it was clear that idazoxan antagonised the reductions in response rate produced by clonidine. Dose-related responding on the idazoxan-associated lever was produced by the anxiolytics buspirone and ipsapirone and by their metabolite MJ 13653 (1 PP), which has previously been shown to be an alpha 2-adrenoceptor antagonist. In general, however, high levels of generalisation occurred with these three compounds only at doses which substantially reduced response rates. These results demonstrate that idazoxan can give rise to a discriminative stimulus which is probably mediated through antagonism at alpha 2-adrenoceptors although the failure of clonidine to block the idazoxan stimulus is difficult to explain. PMID- 2571174 TI - Neuroleptic-induced akathisia: a role for MSH peptides. PMID- 2571176 TI - Serological survey for viral diseases in the Cayo Santiago rhesus macaque population. AB - The free-ranging population of rhesus monkeys (Macaca mulatta) on Cayo Santiago was sero-surveyed for human measles, simian virus 40, B virus (Herpes simiae), rhesus cytomegalovirus, human and simian retroviruses and encephalomyocarditis virus to determine the prevalence of these viruses in the colony. The results of this study indicate that the colony is free of SV40, HTLVIII (HIV-1), STLVIII (SIV) and SRV1; has a low prevalence of measles and EMCV; and high prevalence rates for B virus, CMV and HTLVI. PMID- 2571177 TI - Medetomidine--a novel alpha 2-adrenoceptor agonist: a review of its pharmacodynamic effects. AB - 1. The pharmacodynamic effects of medetomidine, a novel alpha 2-adrenoceptor agonist, are reviewed. 2. In receptor binding experiments, and in isolated organ preparations medetomidine shows high specificity and selectivity to alpha 2 adrenoceptors. Its alpha 2/alpha 1 selectivity ratio is 1620 compared to 220 of clonidine. It is a highly potent full agonist at alpha 2-adrenoceptors, a fact that also distinguishes it from clonidine. 3. Medetomidine induces a dose dependent decrease in the central release and turnover of norepinephrine (NE) measured as changes in metabolite concentrations or using pharmacological intervention techniques. 4. The selectivity, specificity and potency of medetomidine is further supported by various in vivo experiments showing dose dependent hypotensive, bradycardic, sedative, anxiolytic mydriatic, hypothermic and analgesic effects. 5. The pharmacological, neurochemical and behavioral effects of medetomidine can be inhibited by prior, simultaneous or subsequent administration of selective and specific alpha 2-antagonists. 6. In humans medetomidine is well-tolerated and pharmacodynamic effects including e.g. dose dependent decrease of vigilance, blood pressure, heart rate, salivary secretion and plasma NE are compatible with an agonistic action at alpha 2-adrenoceptors. PMID- 2571175 TI - A role for the mesolimbic dopamine system in the psychostimulant actions of MDMA. AB - Methylenedioxymethamphetamine (MDMA) is a phenylethylamine with a chemical structure that resembles both the amphetamines and mescaline and has both stimulant and perception altering properties. The stimulant properties of MDMA were assessed in photocell cages designed to measure locomotor activity in rats. MDMA, over a range of doses (2.5-10.0 mg/kg, SC) produced locomotor hyperactivity which lasted up to 4 h. Further studies examined the role of the mesolimbic dopamine system in the hyperactivity induced by MDMA. 6-Hydroxydopamine lesions of the Nucleus accumbens attenuated the locomotor response produced by MDMA. The well characterized attenuation of the locomotor response produced by amphetamine was also demonstrated in the same rats. The present study demonstrates similarities in the stimulant properties of MDMA and amphetamine, and also suggests that as with amphetamine, the locomotor activation associated with MDMA may involve the presynaptic release of dopamine in the region of the Nucleus accumbens. However, MDMA may have a more unusual pharmacological profile because of its longer duration of action, neurotoxic potential, and differences in the qualitative aspects of its psychoactive effects. PMID- 2571178 TI - Failure of antipsychotic drug dose to explain abnormal diurnal weight gain among 129 chronically psychotic inpatients. AB - 1. The diurnal weight gain was found to be abnormal among 129 chronically psychotic inpatients. 2. The patients were weighed at 7 a.m. and 4 p.m. weekly for three weeks. We normalized the diurnal weight gain (NDWG) as a percentage by subtracting the 7 a.m. weight from the 4 p.m. weight, multiplying the difference by 100, and then dividing the result by the 7 a.m. weight. 3. NDWG was 2.2 +/- 1.5 percent for 87 male patients compared (p less than .0001) with .53 +/- .41 for 14 male controls. 4. NDWG was 1.8 +/- 1.0 percent for 42 female patients compared (p less than .0001) with .49 +/- .30 for 15 female controls. 5. Seventy percent of male and female patients had NDWG values greater than two standard deviations above the mean values of controls. 6. Differences in age, sex, morning weight, antipsychotic drugs, lithium, carbamazepine, phenytoin, blood pressure, and pulse did not explain these findings. PMID- 2571179 TI - Role of dopamine receptors in gastrointestinal motility. AB - Apomorphine (3.5 and 10 mg/kg, s.c.) and morphine (1.3 and 10 mg/kg, s.c.) produced a dose-dependent decrease in gastrointestinal transit of charcoal dust in rats. The involvement of dopamine in such a constipatory effect was found to be mediated to a greater extent by DA2 than by DA1 receptors, using a specific DA1 antagonist (SCH 23390) and DA2 antagonists (alizapride, domperidone). The decrease in apomorphine (50 micrograms) and dopamine (100 and 200 micrograms) induced gastrointestinal transit after intracisternal administration was antagonized by haloperidol. These results provide evidence for the involvement of dopamine receptors in the constipatory effects of apomorphine and morphine in rats. PMID- 2571180 TI - Effects of calcium channel blockers and ketotifen on beta 2 adrenergic receptor regulation in intact human lymphocytes. AB - Effects of three calcium channel blockers (nifedipine, NIF; verapamil, VER and diltiazem, DIL) and one antiallergic drug, ketotifen (KET) on isoproterenol (ISO) induced beta 2 adrenergic receptor desensitization and resensitization have been studied with intact human lymphocytes in vitro. While incubation of lymphocytes with 1.0 microM NIF, VER, or DIL alone neither changed [3H]-CGP-12177 binding (Bmax) nor increased cAMP responsiveness to ISO, they all partially prevented 50 nM ISO-induced beta 2 adrenoceptor-adenylate cyclase uncoupling. VER and DIL also partially prevented beta 2 adrenoceptor down-regulation, but NIF did not. In contrast, 10 microM KET not only diminished ISO-induced desensitization but also accelerated subsequent resensitization. KET alone also induced an up-regulation of cell surface beta adrenergic receptors. The effects of these compounds on beta 2 adrenoceptor regulation may partially explain their potentiation of ISO-induced cAMP accumulation in lymphocytes. PMID- 2571181 TI - [Hyperventilation syndrome; pathophysiology and treatment]. PMID- 2571182 TI - [The concept of neuroimmunology and human neuropathology]. PMID- 2571183 TI - Pituitary gland: neuropeptides, neurotransmitters and growth factors. AB - The hypothalamus receives neuronal afferents from numerous sources including inputs from limbic structures, such as the amygdala and hippocampus, and from brainstem regions involved in the regulation of the cardiovascular system and other autonomic functions. These afferents using a vast array of neurotransmitters and neuropeptides influence the activity of the hypothalamic neurons which synthesize and secrete the hypothalamic releasing and release inhibiting factors into the hypophyseal portal circulatory system. The afferents can modulate the activity of the hypothalamic neurons by forming synapses on the neuronal cell body, on the nerve terminals in the median eminence or both. The chemicals most frequently used as neurotransmitters are the biogenic amines, including the catecholamines (norepinephrine, dopamine and epinephrine), serotonin, acetylcholine and gamma-aminobutyric acid (GABA). The stimulatory influence of norepinephrine, serotonin, and acetylcholine on the secretion of corticotropin (ACTH) in rodents and man will be discussed, whereas GABA exerts an inhibitory effect on the secretion of ACTH in both man and rodents. These effects appear to be mediated by changes in the secretion of the corticotropin-releasing hormone (CRH) and vasopressin into the hypophyseal portal circulation. Numerous neuropeptides appear to alter the secretion of ACTH in the rat. We will discuss the stimulatory actions of neuropeptide Y (NPY), angiotensin II, and peptides of immune cell origin on the secretion of ACTH and CRH. The opioid peptides inhibit the secretion of CRH into the portal blood, however, they exert a potent stimulatory effect on prolactin secretion in the rat and man.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571184 TI - Effect of low-dose exogenous secretin and somatostatin on pentagastrin-stimulated gastric acid secretion in man. AB - The inhibitory effect of intravenous infusion of secretin (0.05 CU kg-1h-1) and somatostatin (60 pmol kg-1h-1), given alone or in combination, on pentagastrin stimulated (100 ng kg-1 h-1) acid secretion was studied in 10 healthy subjects. Secretin inhibited acid secretion by 54% (p less than 0.05), and somatostatin inhibited by 70% (p less than 0.05). The combined infusion of secretin and somatostatin decreased acid output by 64% (p less than 0.05). The differences between the three groups were not significant (p greater than 0.05). Median plasma concentrations of secretin and somatostatin were of the same magnitude as seen after duodenal acidification. The present study did not demonstrate any interaction between secretin and somatostatin in the inhibition of gastric acid secretion. PMID- 2571185 TI - Antibodies to beta 2-microglobulin, the light-chain component of class I histocompatibility molecules, provide an activation signal to human lymphocytes. AB - Antibodies to beta 2-microglobulin (beta 2m), the light-chain component of class I histocompatibility antigens, provide a strong co-stimulatory signal to human lymphocytes in the presence of phorbol ester. This activation signal requires a high concentration of antibody, whereas the effect on responses to a mitogenic lectin is exclusively inhibitory over a broad dose range. A rabbit polyclonal and a mouse monoclonal antibody to beta 2m were similarly co-stimulatory. Antibody to beta 2m was co-stimulatory with phorbol ester, but not calcium ionophore, suggesting that class I antigens play a role in the initiation of the 'first' signal of a recently described two-signal model of lymphocyte activation. PMID- 2571186 TI - Selective killing of CD4+ and CD8+ cells with immunotoxins containing saporin. AB - Saporin, a ribosome-inactivating protein from the seeds of Saponaria officinalis, was covalently linked to an anti-CD4 monoclonal antibody. The resulting immunotoxin at 10(-9)M concentration was toxic to CD4+ lymphocytes without affecting other cells. Selective elimination of CD4+ and CD8+ cells was also obtained with murine monoclonal anti-CD4 and anti-CD8 antibodies and an immunotoxin consisting of saporin linked to an anti-mouse IgG antibody. PMID- 2571187 TI - Synergism between HIV gp120 and gp120-specific antibody in blocking human T cell activation. AB - The human immunodeficiency virus (HIV) binds to CD4-positive cells through interaction of its envelope glycoprotein (gp120) with the CD4 molecule. CD4 is a prominent immunoregulatory molecule, and chronic exposure to antibody against CD4 (anti-CD4) has been shown to cause immunodeficiency in mice. T cell-dependent in vitro immune responses can also be inhibited by anti-CD4. Experimental findings reported here indicate that CD4-bound gp120 attracts gp120-specific antibodies derived from the blood of HIV-seropositive individuals to form a trimolecular complex with itself and CD4. Thus targeted to CD4, the gp120-specific antibody functions as an antibody to CD4; it cross-links and modulates the CD4 molecules and suppresses the activation of T cells as measured by mobilization of intracellular calcium (Ca2i+). The synergism between gp120 and anti-gp120 in blocking T cell activation occurs at low concentrations of both components. Neither gp120 nor anti-gp120 inhibits T cell activation by itself in the concentrations tested. PMID- 2571189 TI - Mechanism of membrane anchoring affects polarized expression of two proteins in MDCK cells. AB - The signals that direct membrane proteins to the apical or basolateral plasma membrane domains of polarized epithelial cells are not known. Several of the class of proteins anchored in the membrane by glycosyl-phosphatidylinositol (GPI) are expressed on the apical surface of such cells. However, it is not known whether the mechanism of membrane anchorage or the polypeptide sequence provides the sorting information. The conversion of the normally basolateral vesicular stomatitis virus glycoprotein (VSV G) to a GPI-anchored protein led to its apical expression. Conversely, replacement of the GPI anchor of placental alkaline phosphatase with the transmembrane and cytoplasmic domains of VSV G shifted its expression from the apical to the basolateral surface. Thus, the mechanism of membrane anchorage can determine the sorting of proteins to the apical or basolateral surface, and the GPI anchor itself may provide an apical transport signal. PMID- 2571188 TI - The protein kinase domain of the ANP receptor is required for signaling. AB - A plasma membrane form of guanylate cyclase is a cell surface receptor for atrial natriuretic peptide (ANP). In response to ANP binding, the receptor-enzyme produces increased amounts of the second messenger, guanosine 3',5' monophosphate. Maximal activation of the cyclase requires the presence of adenosine 5'-triphosphate (ATP) or nonhydrolyzable ATP analogs. The intracellular region of the receptor contains at least two domains with homology to other proteins, one possessing sequence similarity to protein kinase catalytic domains, the other to regions of unknown function in a cytoplasmic form of guanylate cyclase and in adenylate cyclase. It is now shown that the protein kinase-like domain functions as a regulatory element and that the second domain possesses catalytic activity. When the kinase-like domain was removed by deletion mutagenesis, the resulting ANP receptor retained guanylate cyclase activity, but this activity was independent of ANP and its stimulation by ATP was markedly reduced. A model for signal transduction is suggested in which binding of ANP to the extracellular domain of its receptor initiates a conformational change in the protein kinase-like domain, resulting in derepression of guanylate cyclase activity. PMID- 2571190 TI - [Dynorphin and atriopeptin may be involved in the depressor mechanism of clonidine]. AB - Intravenous injection (iv) of clonidine significantly induced depressor and bradycardiac responses in rats. These responses were found to be blocked by iv alpha-adrenoreceptor blocker phentolamine, intraperitoneous injection (ip) of naloxone (10mg/kg) or iv antidynorphin-IgG(80 microliters). Furthermore, clonidine (iv) induced a significant increase of urine volume, which was also partly blocked by naloxone (10mg/kg). A dramatic increase of rat plasma immunoreactive atriopeptin induced by clonidine (iv) was partly antagonised by pretreatment with naloxone (4mg/kg) or phentolamine (10mg/kg). These results show that dynorphin and atriopeptin may be involved in the depressor mechanism of clonidine. PMID- 2571191 TI - Fingering the trigger for neurotransmitter secretion: studies on the calcium channels of squid giant presynaptic terminals. PMID- 2571192 TI - [Therapeutic aspects of hiatal hernia]. PMID- 2571193 TI - [Ergonomic analysis of the work sites and living conditions of the crew on board the B-418/111 t/h "Bonito" PPD and UR "Gryf" fishing trawlers]. AB - The paper is an ergonomic study of the sanitary and living conditions at the working place of fishers on long-distance fishing vessels, such as the trawler B 418/111 t/h "Bonito". The research was performed during the trip in the region of the Falkland Islands (Malvinas). Prophylactic examinations were carried out during 162 days on 77 members of the crew and the hygienic-sanitary conditions on the vessel were analysed. The ergonomic research included all the working places of the members of the crew (fishers on the deck, staff of the industrial processing rooms staff of the machine-rooms and attendant staff) and the following parameters were analysed according to the principles of ergonomics: the characteristics of working places and working conditions, the environmental factors, the anthropometric data of the crew and the physical and psychic stress, for the purpose of optimizing the working conditions according to the physical psychic peculiarities of the members of the crew. PMID- 2571194 TI - [Sulfasalazine in rheumatoid arthritis]. PMID- 2571195 TI - [Drug interactions with calcium inhibitors in man]. AB - The most common interactions concern cardiovascular drugs. The combination of calcium antagonists (CA) and beta-blockers is more effective than single-agent therapy in stable effort angina and hypertension. But there is an increased risk of hemodynamic or electrophysiological side effects in patients with left ventricular or sinus dysfunction, or disturbances of conduction. Pharmacokinetic interactions have been observed in particular with verapamil (VE) which increases propranolol bioavailability. VE increases the T1/2 of elimination and plasma digoxin concentration following single or prolonged administration. The primary mechanism appears to be renal. These modifications increase the risk of digitalis intoxication. Diltiazem (DTZ) inconsistently increases steady state plasma digoxin levels. In healthy subjects, nifedipine (NF) increases plasma digoxin concentrations and decreases digoxin renal clearance. These findings have not been observed in patients with heart failure. NF therefore leads to less marked modifications in digitalis pharmacokinetics than do VE and DTZ. Nitrendipine and nicardipine interact only slightly with digoxin, and consequently there are no pharmacodynamic effects. In healthy subjects, VE increases quinidine t1/2 and markedly decreases its metabolic clearance. Conversely, quinidine increases plasma NF levels. The primary CA are extensively metabolized by liver microsome oxidases. These result in interactions with the drugs that are also metabolized by these enzymes, or able to modify their activity. VE and DTZ decrease antipyrine and carbamazepine clearance. VE, DTZ and nicardipine lead to a marked increase in plasma ciclosporin levels. Cimetidine, but not ranitidine, increases plasma NF levels. The effects on VE are controversial. Prolonged rifampicin treatment decreases plasma VE levels. PMID- 2571196 TI - [Determination of the optimal dose of bromazepam in the elderly]. AB - The aim of our study was to determine in patients over 70 years, the optimal dose of Bromazepam. Seven patients were administered a unique dose of Bromazepam (3 mg), then repeated doses (1.5 mg) every 12 hours for 9 days. Cmax was 42.5 +/- 12 ng/ml in Tmax ranged from 2 to 12 hours after the first dose-concentrations of Bromazepam at steady-state were between 79 and 157 ng/ml. Half life was 30.5 +/- 14.7 hrs for the unique and the repeated doses. Sleeping was noted for concentrations over 40 ng/ml. The dose of 1.5 mg bid is recommended in old patients. PMID- 2571197 TI - A deficiency in A alpha chain's N-terminal alanine residue as a major cause of the slow coagulation of fetal fibrinogen. AB - To clarify the mechanism which causes the coagulation time of fetal fibrinogen to be longer than that of adult fibrinogen, the N-terminal amino acid residues of fetal fibrinogen were analyzed. The results showed that the amount of A alpha chain's N-terminal alanine residues in fetal fibrinogen was only 54% of that in adult fibrinogen. When the amount of A alpha chain's N-terminal residual alanine in adult fibrinogen was decreased to 57% of the normal level by digestion with aminopeptidase M, the adult fibrinogen yielded a prolonged thrombin time and a retarded release rate for fibrino-peptides A and B, both values approximating to those of fetal fibrinogen. The results suggest that the deficiency in alanine residue at the N-terminus of the A alpha chain is a major cause of the slow coagulation of fetal fibrinogen. PMID- 2571198 TI - Differences among beta-adrenoceptor blocking drugs in modifying platelet aggregation and arachidonic acid liberation under thrombin stimulation. AB - The dose-dependent inhibition of thrombin stimulated platelet aggregation due to beta-adrenoceptor blocking drugs followed the rank order of potency: propranolol greater than alprenolol greater than metipranolol and correlated with arachidonic acid (3H-AA) liberation. Atenolol which slightly potentiated stimulated aggregation increased also the liberation of 3H-AA from membrane phospholipids of isolated platelets. Stimulation of platelets resulted in decreased 3H-AA incorporation into phosphatidylcholine, phosphatidylinositol and phosphatidic acid and increased incorporation into phosphatidylethanolamine and phosphatidylserine. Alprenolol, metipranolol and propranolol enhanced the incorporation of 3H-AA into phosphatidylcholine of stimulated platelets. PMID- 2571200 TI - Release of lipoprotein lipase and hepatic triglyceride lipase in rats by heparin and other sulphated polysaccharides. AB - The ability of parenteral heparin to release the capillary-bound enzymes lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) into the circulation is shared by several other sulphated polysaccharides. The lipase releasing activity in rats of two unfractionated heparins has been compared with that of characterised preparations of other glycosaminoglycans and of two partially synthetic polysaccharides. The results suggest that whilst both molecular weight and degree of sulphation are important in determining the potency, there is also some specific structural element associated with the heparin class. The clearance of lipases was also investigated. The half-lives of LPL (29 mins) and of HTGL (36 mins) did not appear to be affected by the nature of the releasing agent. PMID- 2571199 TI - Human mononuclear phagocyte transglutaminase activity cross-links fibrin. AB - The physiologic function of the monocyte transglutaminases is not known. In this study, we detected Factor XIII A-subunit antigen and "tissue" transglutaminase antigen in human monocytes by polyacrylamide gel electrophoresis and immunoblotting techniques. Flow cytometric analysis demonstrated that 27% and 49% of the total Factor XIII antigen in monocytes and human peritoneal macrophages, respectively, are expressed on the surface of the cells. Monocytes maintained in culture for 8 days had a 4-fold increase in Factor XIIIa activity and a 3.2-fold increase in the amount of Factor XIII antigen/mg cell protein. However, there was no increase in the "tissue" transglutaminase activity or antigen levels in cultured monocytes. In addition, we identified a Factor XIII deficient individual who does not express Factor XIII activity or antigen in plasma, platelets, monocytes, lymphocytes or erythrocytes. Intact monocytes from normal donors were able to cross-link fibrin formed in the plasma from the Factor XIII deficient individual. This suggests that transglutaminase activity expressed by peripheral blood monocytes may play a physiologic role in cross-linking fibrin during blood clotting or inflammation. PMID- 2571201 TI - A fatal jellyfish envenomation by the Portuguese man-o'war. AB - The documented case of a human fatality resulting from envenomation by Physalia physalis is presented. Although this case is only the second scientifically recorded fatality, several similar cases have been suspected. PMID- 2571202 TI - Interleukin 2 receptor expression on peripheral blood lymphocytes in association with renal allograft rejection. AB - Periodic assay of IL-2 receptor expression on the surfaces of peripheral blood lymphocytes might provide information predictive of in vivo immunologic events. This study compares two methods of determining IL-2 receptor expression after renal transplantation in cynomolgus monkeys. The first utilized single color staining of peripheral blood mononuclear cells with mouse anti-human IL-2 receptor monoclonal antibody followed by a fluorescein-labeled goat anti-mouse IgG antibody. Epics C cell sorter windows were set to count cells of the size and granularity of normal lymphocytes. The second utilized two-color staining with fluorescein-labeled anti-IL-2 receptor antibody, combined with phycoerythrin labeled anti-CD4 antibody or with phycoerythrin-labeled anti-CD8 antibody. Two color staining allowed the sorter windows to be enlarged to count all mononuclear cells, regardless of size or granularity, without introducing the contaminating effects of monocytes. Data obtained from single-color staining showed no consistent or significant expression of the IL-2 receptor on peripheral lymphocytes in association with the rejection process. Data obtained from two color staining revealed an increase of IL-2 receptor expression on peripheral T cells of at least 10% from the postoperative baseline, which preceded the creatinine rise from allograft rejection in 13 of 13 animals. Increases in IL-2 receptor expression on T cells were not specific to rejection, however. Some animals in which treatment produced a delay of rejection showed a transient rise in IL-2 receptor expression around post-transplant day 5, which was not followed by a rise in creatinine. The two-color staining technique described provides a sensitive means of detecting IL-2 receptor expression in vivo and documents the association of increases in IL-2 receptor expression on T cells with rejection. PMID- 2571203 TI - Differential IL-2 receptor expression in renal allograft recipients treated with an anti-IL-2-receptor antibody. AB - Patients were entered into a randomized trial of prophylaxis for renal allograft rejection by the administration of an anti-human IL-2 receptor antibody, anti Tac, during the first ten days posttransplant. Interleukin-2 receptor (IL-2 R) expression was measured using two anti-IL-2 R monoclonal antibodies (moAbs), anti Tac and 1HT4-4H3. These two antibodies recognize closely spaced epitopes on the 55 kD chain of the IL-2 R. IL-2 R expression was examined on peripheral blood small lymphocytes in three groups of patients who received: (A) cyclosporine CsA and prednisone for baseline immunosuppression (n = 9); (B) anti-Tac with CsA and prednisone as baseline immunosuppression (n = 12); and (C) anti-Tac with azathioprine and prednisone as baseline immunosuppression (n = 5). We found that large numbers of T cells express IL-2 receptors despite the presence of anti-Tac (average of IL-2 R-positive cells at day of peak IL-2 R expression 56.0 +/- 20.8% in group A, 65.2 +/- 26.6% in group B, 21.0 +/- 7.4% in group C). IL-2 R expression did not correlate with clinical activity, and the presence or accessibility of epitopes on the same 55 kD chain varied dramatically from patient to patient. PMID- 2571204 TI - Differential regulation of activation-associated receptor expression on CD4- and CD8-positive T lymphocytes by allosensitized suppressor T cells. AB - We have demonstrated by two-color flow cytometry that allosensitized human suppressor T cells thwart progression of alloactivated T cells into the G1B stage of the cell cycle, and consequently prevent the expression of transferrin receptors on the majority of alloreactive MLR responder cells (P less than 0.005). Suppressor cells inhibit transferrin as well as interleukin-2 receptor expression on both CD4- and CD8-positive responder cells. The addition of 20 U/ml of recombinant interleukin-2 at the initiation of suppressor coculture bioassays completely neutralizes the suppressive effect of suppressor T cells exerted on CD8-positive responder T cells, but does not restore expression of transferrin and interleukin-2 receptors on CD4-positive T cells. Hence, CD4-positive T cells are the direct target of the suppressor cells, while inhibition of transferrin and interleukin-2 receptor expression on CD8-positive T cells results from reduced levels of interleukin-2 in suppressor cell-regulated cultures. In suppressor cell modified mixed lymphocyte reactions supplemented with exogenous interleukin-2 CD8+/CD28- responder cells proliferate preferentially. It is apparent that mixed lymphocyte reactions supplemented with allosensitized suppressor cells cultured in an interleukin-2-rich environment create a milieu that permits the preferential expansion of T cells with suppressor effector phenotype. This in vitro system provides a convenient model in which to cultivate activated CD8+/CD28- T cells. PMID- 2571205 TI - Modulation of the major histocompatibility complex antigen and the immunogenicity of islet allografts. AB - Modulation of major histocompatibility complex (MHC) antigen by parenchymal cells and "passenger leukocytes" is a common feature of allograft rejection. To assess its significance we have examined the fate of antigen-presenting cell (APC) depleted pancreatic islet allografts subsequent to increasing their expression of MHC antigens by in vitro exposure to the lymphokine interferon-gamma (gIFN). While most untreated grafts survived indefinitely, gIFN-exposed grafts were acutely rejected. Using in vitro islet cell-lymphocyte coculture assays, we attempted to dissect the underlying mechanism of enhanced islet cell immunogenicity resulting from gIFN treatment. We determined that gIFN exposure did not affect the capacity of islet cells to serve as APC for T lymphocytes, however islet cell exposure to gIFN was associated with enhanced vulnerability to allogeneic cytotoxic T lymphocyte (CTL) lysis in vitro by an CD5+ (OX-19+), CD8+ (OX-8+), CD4- (W3/25-), class I-restricted CTL. On the basis of these findings, we conclude that antigenic modulation can be a decisive factor in the survival of engrafted tissues by augmenting the interaction of the graft antigens with cytolytic effector T lymphocytes. PMID- 2571206 TI - Specific recognition of human leukemic cells by allogeneic T cell lines. AB - Clinical and experimental data suggest a role for the immune response in preventing leukemic relapses following allogeneic bone marrow transplantation the graft-versus-leukemia (GVL) effect. In the context of an allogeneic BMT, a number of different immune mechanisms mediated by donor cells may be responsible for the GVL effect. We have approached this question by using limiting dilution cultures of alloactivated human lymphocytes to analyze the in vitro allogeneic cytolytic response against fresh allogeneic leukemia. Initial results in the limiting dilution assays with split culture analyses demonstrated frequent alloreactive cytolytic T lymphocyte precursors that destroyed remission peripheral blood lymphocytes and leukemic cells from the allogeneic leukemic patient. These assays also demonstrated frequent lymphokine-activated killer (LAK) cell precursors that lysed both the LAK sensitive Daudi line and the allogeneic leukemia. In these experiments, isolated cultures also showed cytolytic activity directed against the allogeneic leukemic blasts without activity against remission PBL, or the LAK sensitive Daudi cell line. Two T cell lines (ABL1 and ABL2) isolated from an LDA, demonstrated this form of specificity, mediating destruction specifically against the allogeneic acute lymphoblastic leukemic cells. Both cell lines ABL1 and ABL2 were CD3+, TCR alpha beta +, and CD4+. These 2 cell lines mediated little or no cytotoxicity against a large panel of other targets tested (natural killer sensitive and resistant cell lines, allogeneic PBL, and allogeneic fresh leukemic blasts). Antibody-blocking experiments revealed a role for the CD3-TCR receptor of both cell lines in lysis of leukemic cells; the CD4 and MHC class II molecules were clearly involved in the lysis by the ABL1 cell line. Specificity of recognition for the allogeneic leukemic blasts was further confirmed by unlabeled target competitive inhibition studies. The mechanism of the preferential lysis of leukemia by the alloactivated T cell lines described in this paper remains uncertain. Nevertheless, these leukemic-specific populations provide a means by which the human GVL effect may be further studied in vitro. PMID- 2571207 TI - The immunosuppressive mechanism of total lymphoid irradiation. I. The effect on IL-2 production and IL-2 receptor expression. AB - Total lymphoid irradiation is an effective immunosuppressive therapy used to prepare patients for organ transplantation and to treat several autoimmune diseases. We have used the mouse model to investigate the mechanism of TLI induced immunosuppression. Mice were irradiated with 250-rad daily fractions to a total dose of 3500 rads. Splenocytes from control and TLI-treated mice were analyzed for IL-2 production, IL-2 receptor expression after mitogen stimulation, and percent L3T4 positive cells. At two weeks following the completion of TLI, IL 2 production from whole spleen populations had decreased 90% compared with controls (TLI mean IL-2 units = 25 +/- 9.0, control mean = 277 +/- 104). IL-2 receptor expression on splenocytes following Con A stimulation was decreased 80% compared with controls (TLI mean percentage of IL-2 receptor expressing cells = 16.2 +/- 4.1, control mean = 82.0 +/- 7.5). L3T4+ cells that expressed IL-2 receptor were also decreased following TLI (TLI mean = 4.4 +/- 1.4, control mean = 22.0 +/- 3.8), as were proliferative responses to Con A and PHA. Addition of recombinant mouse IL-2 did not restore IL-2 receptor expression or proliferative responses. Whole TLI-treated splenocytes did not suppress IL-2 production or proliferative responses of normal splenocytes. These immunologic abnormalities recovered over time, and by 8 weeks post TLI IL-2 production, IL-2 receptor expression, L3T4+ cell numbers and proliferative responses had returned toward normal. These results suggest that TLI therapy transiently depletes IL-2 producing, IL-2 receptor expressing, and mitogen responsive lymphocytes. The immunosuppression is not mediated through a suppressor cell and is independent of IL-2 production. PMID- 2571208 TI - Evidence that initial T cell depletion suppresses antiantibody formation and preserves immunosuppression after polyclonal ATG. PMID- 2571209 TI - Determination of beta-adrenoceptors and cAMP in muscle from normal and splayleg Belgian Landrace pigs. AB - beta-Adrenoceptors were identified and characterized by [3H]dihydroalprenolol ([ 3H]DHA) binding experiments in muscle membrane preparations from piglets. The [3H]DHA binding was rapid, reversible and stereoselective. Catecholamines competed for specific binding with a rank order of potency (-) isopropylnoradrenaline greater than (-)-epinephrine much greater than (-) norepinephrine, indicating a beta 2-subtype of adrenoceptor. Saturation binding experiments with [3H]DHA showed no significant difference for either the number of binding sites or the equilibrium dissociation constants in normal and splayleg pigs. Adenylate cyclase assays indicated that the basal adenylate cyclase activity and the prostaglandin E1 (PGE1)-, (-)-isopropylnoradrenaline (ISO)-, 5' guanylyl-imidodiphosphate (GppNHp)- and sodium fluoride (NaF)-stimulated values were not significantly different in normal and splayleg pigs. In both groups, PGE1 did not affect basal activity, whereas ISO and GppNHp stimulated adenylate cyclase activity significantly (p less than 0.001) to about 40% above basal level. NaF induced a significant (p less than 0.001) increase of cAMP in normal and splayleg pigs amounting to 48% and 61% respectively. Significant correlations between absolute adenylate cyclase responses to ISO (r = 0.83***), NaF (r = 0.72**) and GppNHp (r = 0.61*) and basal activity in the splayleg pigs were the most striking findings. In contrast, these correlations could not be detected in the normal pigs. Whether or not this observation reflects an alteration in the signal transduction system needs to be further investigated. To the best of our knowledge this is the first biochemical study which relates an altered beta adrenoceptor function and porcine splayleg. PMID- 2571210 TI - Coexistence of NMDA and non-NMDA receptors on turtle horizontal cells revealed using isolated retina preparations. AB - Effects of glutamate and its agonists on horizontal cells appeared diversely when studied in turtle eyecup preparations. Consistent results were obtained when the isolated retina preparations were used. Not only kainate and quisqualate but also N-methyl-D-aspartate (NMDA) caused a sustained depolarization and light-evoked responses were suppressed for as long as these agonists were superfused. A selective antagonist of NMDA, 2-amino-5-phosphonovalerate (APV), hyperpolarized horizontal cells and reduced their light-evoked responses. These results indicate the coexistence of NMDA and non-NMDA receptors on turtle horizontal cells. PMID- 2571211 TI - Low AIDS attack rate among Dutch haemophiliacs compared to homosexual men: a correlate of HIV antigenaemia frequencies. AB - A cohort of 180 haemophiliacs followed between 1983 and 1986 and a cohort of 961 homosexual men followed between 1984 and 1986 were compared for the prevalence and incidence of HIV-1 antibody (HIV-1-Ab) seropositivity, the incidence of AIDS related complex (ARC) and AIDS and the prevalence and incidence of serological and immunological markers for HIV-related disease progression. Among the haemophiliacs 23 (12.8%) patients were HIV-1-Ab seropositive at entry and 20 (12.7%) of the remaining 157 seroconverted for HIV-1-Ab during follow-up. Of the homosexual men 238 (24.8%) were HIV-1-Ab seropositive at entry and 68 (9.4%) of the 723 at entry seronegatives seroconverted during follow-up. Clinical follow-up of the HIV-1-Ab seropositive and seroconverted men was 59 months in the haemophiliac cohort and 60 months in the homosexual cohort. Among the HIV-1-Ab seropositive and seroconverted haemophiliacs and homosexual men the cumulative ARC/AIDS incidence was 2 and 18%, respectively. Occurrence of HIV-1-antigenaemia was more frequent among seropositive and seroconverted homosexual men (28%) than among haemophiliacs (7%) (p = 0.001). The groups did not differ significantly for the absence or loss of anti-HIV core antibodies or the occurrence of low CD4+ cell numbers. These data indicate a slower progression of HIV-related disease in seropositive haemophiliacs compared to seropositive homosexual men. PMID- 2571212 TI - Application and evaluation of dissemination model. PMID- 2571213 TI - Permanent brain damage and pertussis vaccination: is the end of the saga in sight? AB - The development and widespread uptake of a safe and efficacious aluminium adsorbed diphtheria-tetanus-pertussis vaccine (DTP) in the United States between 1933 and 1944 led to a gradual decline in whooping cough morbidity and instilled confidence in a vaccination programme which has been effectively maintained for over 40 years. T his contrasts with the turbulent history of pertussis vaccination in the United Kingdom where doubts as to the efficacy of available vaccines delayed their active national promotion until 1957, after which various reports resulted in further doubts over efficacy and safety. In 1974, the mass media became involved in the safety issue when the National Hospital for Sick Children case series of neurological events, which had occurred after DTP vaccination was made core material for a television documentary. The Department of Health and Social Security (DHSS) responded by establishing two retrospective studies of case records of post vaccination adverse events and two prospective studies. One of the latter, the National Childhood Encephalopathy Study (NCES) was regarded as the definitive case control study. A claim for damages, Loveday v Renton and The Wellcome Foundation, heard in the High Court of Justice in London, from early October 1987 until late February 1988 dealt with the general issue of whether, on the balance of probabilities, pertussis vaccine could cause permanent brain damage. The cornerstone of the claim that pertussis vaccine can cause permanent brain damage has always been the apparent clustering of onset of neurological disorders within the first 24-48 h after vaccination. One of the main finds of the NCES, however, which was not divulged in any published report but emerged in the course of the hearing, was that permanent brain damage did not occur within 48 h of DTP vaccination in any child in England, Scotland and Wales from mid-1976 to mid-1979 when 2 million doses of vaccine were used. The NCES, in this respect, completely undermined the evidence provided by various published case series. After a 61 day hearing, High Court judgment was given to the effect that the judge was not satisfied on balance of probability that pertussis vaccine can cause permanent brain damage in young children. Were it not for the fact that The Wellcome Foundation intervened in the action and obtained a court order giving access to the NCES cases records, some crucial information collected by the NCES might not have been brought to light.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2571214 TI - [Clinical and pharmacokinetic aspects of the problem of benzodiazepine derivatives (review of foreign literature)]. PMID- 2571215 TI - [Use of preparations of the benzodiazepine group in the treatment of epilepsy (review of the literature)]. PMID- 2571217 TI - Clorazepat in the treatment of anxiety-depression syndroms in neuroses. PMID- 2571216 TI - Absence of cardio-depressant effects of CGP 17582 B, a new beta-blocking agent, in patients after coronary surgery. AB - The hemodynamic effects of CGP 17582 B, a new cardio-selective beta-blocking agent with moderate intrinsic sympathomimetic activity and minimal effects on myocardial contractility, were studied in patients after cardiac surgery for coronary artery bypass graft. Each patient had been treated preoperatively with beta-blocking agents and had a cineangiographic left ventricular ejection fraction between 40 and 60%. Fourteen patients were randomized to receive either 10 mg of propranolol or 50 mg of CGP 17582 B orally. Both drugs resulted in a significant and a similar decrease in heart rate. However, this was associated with a significant decrease in stroke volume after propranolol but not after CGP 17582 B, so that cardiac output significantly decreased only after propranolol. Thermodilution right ventricular ejection fraction significantly decreased after propranolol but not after CGP 17582 B. Each drug was well tolerated during the 10 following days and the recovery was uneventful in each patient. These results indicate that CGP 17582 B is a promising beta-blocking agent susceptible to reduce heart rate without altering cardiovascular function after cardiac surgery. PMID- 2571218 TI - Controlled cross-over comparison of isofloxythepin and perphenazine in the treatment of schizophrenic psychoses. PMID- 2571219 TI - Visual evoked potentials during the treatment with isofloxythepine. PMID- 2571220 TI - Cross-over comparison of the therapeutic effects of sulpiride and perphenazine in schizophrenic and schizoaffective psychoses. PMID- 2571221 TI - Results of the long term treatment with clozapine. Clinical and electroencephalographic study. PMID- 2571222 TI - Intraindividual comparison of the effect of perphenazine and of the combination of perphenazine and lithium in schizophrenia. PMID- 2571223 TI - Pharmacokinetic properties of haloperidol decanoate. PMID- 2571225 TI - Haloperidol decanoate in the treatment of sexual deviations. PMID- 2571224 TI - Haloperidol decanoate in the treatment of schizophrenia. PMID- 2571226 TI - Disposition and pharmacokinetics of the new neuroleptic drug VUFB 15496 (cloflumide) in rats. PMID- 2571227 TI - Oxyprothepin and clorotepin compared in the hyperkinetic syndrome. PMID- 2571228 TI - Therapeutic results in acute phases of manic depressive and schizophrenic psychoses as possible predictors of the prophylactic effect. PMID- 2571229 TI - Sedative and anxiolytic potency of benzodiazepines: some structure-activity relationships. PMID- 2571230 TI - Effect of RG-tannin on yawning behaviour induced by apomorphine or physostigmine in rats. PMID- 2571231 TI - Animal model of anxiety: interaction of nicergoline and scopolamine in the genetically hypertensive rats. PMID- 2571232 TI - Development of gastrin-and somatostatin-containing cells in the pyloric gland region of the bovine abomasum. AB - The ontogeny of gastrin- and somatostatin-containing cells in the pyloric gland region of the abomasum was studied in bovine fetuses, newborns and adults, using immunohistochemical techniques. A clear-cut developmental increase in the two kinds of endocrine cells could not be observed. The region of the lesser curvature displays more D and G cells than the greater curvature. Cells of both open and closed types are found from the earliest stages examined and continue to be present throughout the complete differentiation of the pyloric mucosa. PMID- 2571233 TI - Sebum secretion in idiopathic Parkinson's disease: effect of anticholinergic and dopaminergic drugs. AB - Sebum secretion was measured on the foreheads of 47 unmedicated patients with idiopathic Parkinson's disease, aged 50-81 years and 80 healthy, matched volunteers. Sebum secretion was significantly higher in men patients aged 50-59 years than in controls; no such difference was observed in women of the same age. There was also no difference between patients and volunteers of either gender older than 60 years of age. The effects of either biperiden, L-Dopa + benserazide (a decarboxylase inhibitor) or bromocriptine treatments on sebum secretion and on 5 parkinsonian signs were evaluated over 127 days. There was a significant reduction of all measures during the treatment with the dopaminergic drugs. The only observable effect with biperiden was reduction of tremor. It is discussed whether the sebum suppressive effect of the dopaminergic drugs may be explained through their central effects. PMID- 2571234 TI - Effects of beta-adrenergic agonists in the parotid gland of the rat--an electrophysiological study. AB - The effects of some beta-adrenergic agonists were studied in the parotid gland of the rat by electrophysiological techniques. In the unoperated gland, isoprenaline caused depolarizations which were slowly developing, long-lasting and of low amplitude. The same response was seen when noradrenaline was combined with alpha adrenoceptor blocking drugs. A greater number of cells responded to this combination than to isoprenaline. After either parasympathetic or sympathetic denervation 1-3 weeks in advance, to induce supersensitivity, the number of cells responding to beta-adrenoceptor stimulating drugs was significantly increased. In the latter case the threshold dose required to evoke a response was also significantly lowered. Atropine did not have any effect on the isoprenaline evoked response. The combined parasympathetic and sympathetic denervation did not further increase the responsiveness. It is concluded that beta-adrenoceptor stimulation in the parotid gland of the rat may cause membrane depolarizations. The response is mediated by beta 1-adrenoceptors. The responsiveness is increased in the denervated gland. Secretory studies have demonstrated a supersensitivity to beta-adrenergic agonists as a result of denervation. On the other hand, beta adrenoceptor stimulation is believed mainly to activate the adenylate cyclase/cyclic AMP system independent of membrane potential changes. It is thus not known if the present 'supersensitivity' is correlated to the increased secretory response earlier demonstrated in this gland. PMID- 2571236 TI - Neuropeptides in the human internal mammary artery. PMID- 2571235 TI - Release and vasoconstrictor effects of neuropeptide Y in relation to non adrenergic sympathetic control of renal blood flow in the pig. AB - The possible involvement of neuropeptide Y (NPY) in sympathetic control of renal blood flow was investigated in the pig in vivo. Exogenous NPY caused renal vasoconstriction with a threshold effect at an arterial plasma concentration of 164 pmol 6(-1). Stimulation of the renal nerves (0.59, 2 and 10 Hz) in control animals evoked rapid and frequency-dependent reduction in renal blood flow and overflow of NPY-like immunoreactivity (NPY-LI) and noradrenaline (NA) from the kidney, suggesting co-release from sympathetic nerves. Following the administration of the alpha- and beta-adrenoceptor antagonists phenoxybenzamine and propranolol, the vasoconstrictor response to exogenous NA was reduced by 98%, whereas that of NPY was unaltered. The response to nerve stimulation with 0.59 Hz was abolished, whereas relatively slowly developing reductions in renal blood flow by 7 and 28% were obtained upon stimulation with 2 and 10 Hz respectively. The nerve stimulation-evoked overflow of NA at 0.59 and 2 Hz, but not at 10 Hz and not that of NPY-LI, was enhanced after adrenoceptor blockade. Twenty-four hours after reserpine treatment (1 mg kg-1 i.v.) the contents of NPY-LI and NA in the renal cortex were reduced by 80 and 98% respectively. Sectioning of the renal nerves largely prevented the reserpine-induced depletion of NPY-LI, but not that of NA. Nerve stimulation of the denervated kidney with 2 and 10 Hz 24 h after reserpine treatment evoked slowly developing and long-lasting reductions in renal blood flow by 6 and 52% respectively. These responses were associated with overflow of NPY-LI, which was similar to and threefold higher than that observed in controls at 2 and 10 Hz respectively, while no detectable overflow of NA occurred. Repeated stimulation with 10 Hz resulted in a progressive fatigue of the vasoconstrictor response and the associated overflow of NPY-LI, giving a high correlation (r = 0.86, P less than 0.001) between the two parameters. It is concluded that NPY is a potent constrictor of the renal vascular bed. Furthermore, although NA is the likely transmitter mediating most of the responses to low to moderate nerve activation under control conditions, the data suggest that NPY may mediate the non-adrenergic reductions in renal blood flow evoked by high-frequency sympathetic nerve stimulation after reserpine treatment. PMID- 2571237 TI - Is cyclic AMP the intra-axonal messenger 'X' mediating 'upstream' control of sympathetic transmitter secretion? PMID- 2571238 TI - Vagally induced hexamethonium-resistant jejunal contractions in the cat. PMID- 2571239 TI - Spatial distribution of coronary capillaries: A-V segment staggering. PMID- 2571240 TI - Influences of different routinely used muscle relaxants on oxygen delivery to and oxygen consumption by the heart during xenon-anesthesia. PMID- 2571241 TI - Cytoprotective effect of isotonic mannitol at low oxygen tension. AB - The beneficial effect of mannitol infusion in postischemic kidneys remains unresolved. Contradictory reports may have originated from at least 3 different mechanisms: a) arterial vasodilation, b) osmotic support of hypoxic cellular volume regulation, and c) scavenging of hydroxyl radicals in the reoxygenation period. To exclude vascular effects we tested at 37 degrees C in hypoxic (PO2 less than 1 mmHg) and reoxygenated isolated tubular cells of rat kidney cortex cellular function, i.e. intracellular K+ accumulation (K+), posthypoxic lactate gluconeogenesis (GNG), loss of membrane-bound tau-Glutamyltransferase (tau GT), and formation of a lipid peroxidation product, malondialdehyde (MDA). Mannitol (M) was added to a Ringer incubation medium in variable concentration either without (hypertonic M) or with omission of equiosmolar amounts of NaCl (isotonic M). K+ and GNG were significantly supported, and tau GT-loss markedly suppressed in a range of 10-50 mmol/l hypertonic and isotonic M. At higher concentrations no improvement (isotonic) or even deleterious effects (hypertonic) of M occurred. Beneficial effects of lower concentrations of M (10 mmol/l) were not correlated to anaerobic glycolysis, and 1 as well as 10 mmol/l M induced a comparable and significant reduction in posthypoxic MDA-formation. This effect was most pronounced when M was only added in hypoxia, indicating leakiness of cellular membranes in hypoxia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571242 TI - Renal involvement in Hantavirus disease. PMID- 2571243 TI - Responses to histamine and selective H2-receptor agonists in lung parenchymal strips from normal and sensitized guinea-pigs. AB - Histamine produces concentration-dependent contractions of lung parenchyma strips obtained from normal and sensitized guinea-pigs. The responsiveness of the sensitized lung strips to histamine was significantly increased compared to normal tissues. Clemizole (0.1 microM) was equally effective as an H1-antagonist in normal (dose-ratio 9.12) and sensitized (dose-ratio 9.77) tissues. The concentration-response curves to histamine were displaced to the left by cimetidine (0.1 microM to 0.1 mM) with similar dose-ratios in normal and sensitized tissues. Cimetidine enhanced maximal responses to histamine only in normal lung strips. The effects of submaximal equieffective concentrations of histamine were augmented to the same extent by cimetidine (0.1 mM) in normal and sensitized tissues. The responses to histamine were not modified by indomethacin (5 microM). The responsiveness and sensitivity of sensitized lung strips to isoprenaline, impromidine, 4-methyl-histamine and dimaprit were not different from those of normal tissues. Cimetidine yielded, as antagonist of dimaprit, similar pA2 values in normal and sensitized tissues. In conclusion, there is no experimental evidence in favour of the existence of an impairment of H2-receptor activity in sensitized airways. Hyperreactivity to histamine is probably due to differences between normal and sensitized tissues with respect to Ca2+ entry and/or intracellular Ca2+ release in response to H1-receptor activation. PMID- 2571244 TI - Rocastine (AHR-11325), a rapid acting, nonsedating antihistamine. AB - Rocastine [AHR-11325, 2-[2-(dimethylamino)ethyl]-2,3-dihydro-4-methylpyrido-[3,2 f]-1,4- oxazepine-5(4H)-thione (E)-2-butenedioate)] is a rapid-acting, potent, nonsedating antihistamine. In guinea pigs challenged with a lethal dose of histamine, rocastine is as effective [based on 1 hr. oral, protective dose (PD50S)] as brompheniramine, chlorpheniramine, pyrilamine, and promethazine and superior to astemizole, diphenhydramine, terfenadine, and oxatomide. Rocastine has a faster onset of action than does terfenadine; rocastine being as effective with a 15 min pretreatment time (PD50 = 0.13 mg/kg) as it is with a 1 hr pretreatment time (PD50 = 0.12 mg/kg), while the 15 min PD50 of terfenadine (PD50 = 44.0 mg/kg) is 22 times greater than the 1 hr PD50 (PD50 = 1.93 mg/kg). Against aerosolized histamine, rocastine was 7.12 x, 2.63 x, and equipotent to pyrilamine in preventing histamine-induced prostration at pretreatment times of 1,3, and 6 hr, respectively. Rocastine protected guinea pigs from collapse induced by aerosolized antigen; rocastine was approximately 36 x more potent (based on 1 hr PD50) than diphenhydramine and as potent as oxatomide and terfenadine. Rocastine did not alter the EEG of cats at doses in vast excess (150x) of its antihistaminic dose nor did it potentiate yohimbine toxicity in mice. Further, rocastine possesses no anticholinergic, antiadrenergic, or antiserotonergic properties in vitro. Rocastine is a selective, nonsedating, H1-antagonist with a rapid onset of action. PMID- 2571245 TI - Histamine receptors in unstimulated pancreatic exocrine secretion of the rabbit. AB - Histamine H2-receptor antagonists cimetidine and oxmetidine, H2-receptor agonist dimaprit, H1-receptor antagonist chlorpheniramine and H1-receptor agonist 2 thiazolylethylamine were tested for their effects on unstimulated pancreatic exocrine secretion in anaesthetized rabbits fitted with an acute pancreatic cannula. Intravenous administration of H1 agonist induces a dose-dependent increase in pancreatic secretion but H1 antagonist have the opposite effects. Intravenous administration of H2 antagonists induces effects similar the ones produced after H1 agonist infusion. The implications of H1 and H2 receptors on exocrine pancreatic secretion are discussed. PMID- 2571247 TI - Delay in emergence of mefloquine resistance in Plasmodium berghei by use of drug combinations. AB - Blood induced Plasmodium berghei infection in Swiss mice was exposed during successive passages to melfloquine alone or melfoquine in combination with dapsone or primaquine or erythromycin, and the level of resistance to melfoquine in four sub-lines was compared at ED90 level. Treatment with mefloquine alone resulted in a 201.14 fold increase in resistance after 21 passages. Use of drug combination (melfoquine + dapsone) delayed the acquisition of resistance as shown by a marginal increase of ED90 by 5.76 fold after 34 passages. Similarly, there was a 17.84 fold increase of resistance after 32 passages involving exposure to mefloquine-primaquine and a 112.28 fold increase after exposure to melfoquine erythromycin combination for 31 passages. The study emphasizes that rational drug combinations should be developed to protect the melfoquine against the emergence of resistance in P. falciparum cases. PMID- 2571246 TI - The effect of azelastine and some other antiasthmatic and antiallergic drugs on calmodulin and protein kinase C. AB - The antiallergic and antiasthmatic drug, azelastine, interacts strongly with calmodulin (but not bovine serum albumin) as determined by an indirect assay; it also moderately inhibited the Ca2+-calmodulin-dependent enzyme bovine brain phosphodiesterase. Ketotifen was less active than azelastine in both assays of calmodulin reactivity and both drugs were less active than the recognized calmodulin inhibitor, W-7. Neither azelastine nor ketotifen had any inhibitory effect on the Ca2+- and phospholipid-dependent protein kinase C. A number of other commonly employed antiallergic and antiasthmatic drugs were essentially inactive in the calmodulin assays and had no or marginal inhibitory effect on protein kinase C. PMID- 2571248 TI - Isoenzyme characterization of Trypanosoma evansi isolated from capybaras and dogs in Brazil. AB - Trypanosoma evansi was seen in blood samples taken randomly from both wild and semi-captive capybaras caught in Mato Grosso do Sul State, Brazil and in sick dogs brought into local veterinary clinics. Trypanosome stocks from capybaras and dogs were significantly different in their patterns of growth in mice, while the trypanosomes from dogs were mostly dyskinetoplastic. By isoenzyme electrophoresis all the trypanosomes were of the most common type of T. evansi found elsewhere. PMID- 2571249 TI - Rickettsial infections of midgut cells are not associated with susceptibility of Glossina morsitans centralis to Trypanosoma congolense infection. AB - Teneral and 30-day old non-teneral Glossini morsitans centralis, from a laboratory-bred colony, were fed on a goat infected with Trypanosoma congolense clone IL 1180. They were then maintained on an uninfected rabbit, and dissected on day 30 after the infected feed. The midgut infection rates were 38.1% and 8.1%, with the mature infection rates of 28.7% and 4.3%, respectively. Electron microscopical examination revealed the presence of rickettsia-like organisms (RLOs) within the mycetomes and the midgut epithelial cells of all the teneral and non-teneral tsetse examined; the RLOs being more numerous in the older tsetse. Also, when the infected feed was given to teneral tsetse, maintained as above and dissected 30 days later, RLOs were observed in the tsetse with mature and immature T. congolense infections, as well as in those tsetse which completely lacked the trypanosomes. It appears that susceptibility of the laboratory reared G.m. centralis to T. congolense infection was not associated with RLOs within the midgut epithelial cells. PMID- 2571250 TI - Assessment of the prevalence and titer of antibodies to a candidate schistosomiasis vaccine molecule, Sj26, in several human serum banks. AB - Using immunoassay and immunoblotting approaches, antibodies to Sj26, a glutathione S-transferase molecule (Mr = 26,000) of Schistosoma japonicum worms that is a vaccine candidate, have been sought in three large human serum banks. For these studies, a near-native recombinant Sj26 molecule produced in Escherichia coli was used, generally in ELISAs. Anti-Sj26 activity was detected readily in a high proportion of the sera at titres below 1:400 and appeared to be largely protein A-binding IgG antibodies. No differences in the prevalence of anti-Sj26 antibodies were noted in sera from entirely normal individuals or those with a variety of parasitic infections, but never exposed to S. japonicum. The stimulus responsible for induction of these low titre, probably low affinity antibodies in humans remains unknown. PMID- 2571251 TI - Immune-complexes-mediated evasion of Plasmodium knowlesi from destruction by macrophages. AB - The role of immune-complexes in the evasion of Plasmodium knowlesi from destruction by macrophages was studied in vitro. Incubation of macrophages with immune-complexes, prepared either by mixing total parasite antigens soluble in culture medium with normal or immune monkey serum, or by polyethylene glycol precipitation of serum from monkeys acutely infected with P. knowlesi, significantly reduced both the pool size of the macrophages that bound parasitized erythrocytes, and the number of parasitized erythrocytes bound per macrophage. Parasitized erythrocytes with mature schizonts were invariably preferred over those containing rings. These observations appear to indicate that during P. knowlesi infection in rhesus monkeys, immune-complexes may inhibit the binding of parasitized erythrocytes with mononuclear phagocytes and thus may enable them to evade the destructive mechanisms mounted by the host. PMID- 2571252 TI - Hypogonadism and ecdysteroid production in Loa loa and Mansonella perstans filariasis. AB - Possible endocrinological repercussions of infection with Loa loa and Mansonella perstans filariae were studied in Gabonese subjects. Microfilaremic males were compared with amicrofilaremic controls. In the infected group 13/105 subjects (12%) presented only abnormally low serum levels of testosterone (less than 4 ng/ml), 25/105 (24%) only abnormally high serum levels of gonadotrophins, FSH (greater than 15 mIU/ml) and LH (greater than 20 mIU/ml), and 22/105 (21%) presented anomalies in both testosterone and gonadotrophin levels. One out of 68 control subjects had 3.6 ng/ml seric testosterone and all had normal levels of gonadotrophins. Ecdysteroids were detected (greater than 0.025 ng/ml) in the serum of 87/97 (90%) microfilaremic subjects (GM 0.123 ng/ml) compared to 12/64 (19%) controls (GM 0.030 ng/ml). Ecdysteroids were detected in the urine of all subjects, infected (GM 8.468 ng/ml) as well as control (GM 1.245 ng/ml). The hormonal perturbations were correlated with the levels of Loa loa microfilaremia but not with those of serum and urinary ecdysteroids. These results demonstrate that microfilaremic subjects often show endocrinal signs of hypogonadism and present appreciable levels of ecdysteroids in serum and urine. A direct role for parasitic ecdysteroids in hypogonadism remains to be demonstrated. PMID- 2571254 TI - Pyrethroid-impregnated bed nets in the malaria control strategy at community level. PMID- 2571253 TI - Immunodiagnosis of sleeping sickness due to Trypanosoma brucei gambiense by detection of antiprocyclic antibodies and trypanosome antigens in patients' sera. AB - Documented sera from 39 Trypanosoma brucei gambiense sleeping sickness patients from Ivory Coast (Cote d'Ivoire) were tested using the Procyclic Agglutination Trypanosomiasis Test (PATT) for the presence of anti-trypanosome antibodies and using an antigen-capture double antibody enzyme-linked immunosorbent assay (ELISA) for the presence of trypanosomal antigens. All 39 sera contained antiprocyclic antibodies and trypanosome antigens whereas 5 control sera did not. The results show that the PATT (for antibody detection) and the double antibody ELISA (for antigen detection) are useful for immunodiagnosis of African sleeping sickness due to T.b. gambiense and that these assays should be simplified for further testing and evaluation in the field. PMID- 2571255 TI - Reduced fertility in mice double-infected with Schistosoma mansoni and Echinostoma revolutum. PMID- 2571256 TI - Application of anti-TNF to Plasmodium vinckei-infected mice is followed by an increase of parasitaemia. PMID- 2571257 TI - Medetomidine as a preanesthetic prior to ketamine-HCL and halothane anesthesia in laboratory beagles. AB - The potent sedative and analgesic effects of medetomidine confirm its potential use as a premedication to general anesthesia in dogs. In this study two different doses of medetomidine were tested as premedication to both ketamine HCl and halothane anesthesia in groups consisting of 4 laboratory beagles each. Cardiopulmonary parameters, anesthetic quality and dose requirements were recorded. Medetomidine was found to have favorable qualities in conjunction with these anesthetics. Atropine prevented the profound bradycardia and sinus arrhythmia seen with medetomidine alone. Cardiopulmonary parameters remained satisfactory in both groups, but no additional benefits could be seen from doses of 40 micrograms/kg medetomidine compared to 20 micrograms/kg, except a significant 30% reduction in halothane requirement. The positive chronotropic and inotropic properties of ketamine restored the medetomidine induced bradycardia and produced a short anesthetic period of 15-30 min depending on the dose of medetomidine. The quality of anesthesia was good in both groups, but recovery was quicker and smoother in the group with 20 micrograms/kg medetomidine. PMID- 2571258 TI - An introduction to the pharmacology of alpha 2-adrenoceptors in the central nervous system. AB - alpha 2-Adrenoceptors mediate a variety of physiological functions in the central nervous system and in peripheral tissues; thus, the use of pharmacological agents either activating or blocking these receptors results in a mixture of responses from several organs and tissues. This may be utilized therapeutically in some situations. In anaesthesiology, alpha 2-adrenergic agonists are already widely used as veterinary sedative-analgesics, and it is now clear that they are also capable of potentiating the effects and reducing the required dosage of other drugs used in anaesthesia (barbiturates, volatile anaesthetics, narcotic analgesics). In addition, they attenuate sympatho-adrenal responses to noxious stimuli encountered during anaesthesia and surgery, providing improved hemodynamic, metabolic and hormonal stability. Recent results from molecular biology, biochemistry and pharmacology point to the existence of distinct alpha 2 adrenoceptor subtypes. The identification of such subpopulations of alpha 2 adrenoceptors may provide new opportunities for the development of subtype selective pharmacological agents with more specific therapeutic actions. PMID- 2571259 TI - Comparison of three different dose regimes of medetomidine in laboratory beagles. AB - Different dose regimes of medetomidine (a potent alpha 2-adrenergic agonist), adding up to a combined dose of 80 micrograms/kg, were administered to laboratory beagles to determine the possible superiority of any particular regime. The study was intended to mimic a clinical situation where sufficient sedative and analgesic effect was not reached with the initial dose and an additional dose would have to be administered. Cardiopulmonary responses were very similar in each dose regime, showing the characteristic properties of single doses of 80 micrograms/kg of medetomidine as reported in the literature. Effective sedative and analgesic duration seemed to be a function of when the largest dose was administered. Adequate additional sedative and analgesic effect could be gained from the second injection at doses of half of the initial one. PMID- 2571260 TI - Medetomidine/ketamine anaesthesia in cats. AB - The medetomidine/ketamine combination was evaluated as an anaesthetic in cats undergoing ovariectomy. It was compared with acepromazine/ketamine, xylazine/ketamine and zolazepam/tiletamine combinations. 60 animals, divided into 6 groups of 10 animals were involved. Medetomidine highly potentiated the anaesthetic effects of ketamine and balanced the two main disadvantages of this drug: weak muscle relaxation and poor analgesia in deep organs. 80 micrograms medetomidine/kg--5 mg ketamine/kg provided a duration of anaesthesia longer than 1 mg acepromazine/kg--10 mg ketamine/kg, and not significantly different from those induced by 1 mg xylazine/kg--10 mg ketamine/kg or 7.5 mg zolazepam/kg--7.5 mg tiletamine/kg. Furthermore, it produced a better muscle relaxation than acepromazine/ketamine and zolazepam/tiletamine combinations. The analgesia in deep organs observed after medetomidine/ketamine administration appeared to be more substantial than that observed after acepromazine/ketamine or xylazine/ketamine combinations. The stimulating effect on heart rate of ketamine compensated the bradycardiac effect of medetomidine. Nevertheless, the administration of 80 micrograms medetomidine/kg--5 mg ketamine/kg resulted as with xylazine/ketamine, in decreasing heart rate whereas acepromazine/ketamine and zolazepam/tiletamine combinations did not exhibit bradycardiac effect. It was concluded that the medetomidine/ketamine combination provided a suitable anaesthesia for cats characterized by rapid induction, good muscle relaxation, good analgesia and bradycardia. PMID- 2571261 TI - Preliminary results on the use of medetomidine-ketamine combinations in the dog. AB - The effects of medetomidine and ketamine by subsequent or simultaneous in injections were studied. ECG, arterial blood gas analyses and mean arterial pressure measurements were done periodically during the 60 min monitoring period. Medetomidine and ketamine induced deep sedation that proved useful for minor operations in clinical work. PMID- 2571263 TI - Medetomidine--ketamine--diazepam anesthesia in the rabbit. AB - Orthopaedic operations were performed in 340 rabbits when developing biodegradable rods for the fixation of fractures. The rabbits were anaesthesized by injecting medetomidine combined with ketamine and diazepam (MKD) subcutaneously. The mortality due to the anaesthesia was zero. Arterial blood gas analyses showed a moderate decrease in the haemoglobin oxygen saturation and compensated respiratory acidosis, which was well tolerated by the animals. It is concluded that MKD anaesthesia is well suited for orthopaedic operations in the rabbit. PMID- 2571262 TI - Uterine motor responses to an alpha 2-adrenergic agonist medetomidine hydrochloride in the bitches during the end of gestation and the post-partum period. AB - Uterine motor responses to an alpha-2 adrenergic agonist, medetomidine hydrochloride in bitches during the end of gestation and the post-parturient period. The electrical activity of the uterine muscular coat in bitches was recorded during the end of the pregnancy and on 2nd, 4th, 6th and 8th day after the parturition, before and after intravenous injections of medetomidine in the doses of 20, 40 and 60 micrograms/kg b.w. The effect of the drug was observed 5 30 s after the injection and it lasted from 20 to 55 min. A small dose of 20 micrograms/kg b.w. caused decrease of electrical activity of the pregnant uterus. The doses of 40 and 60 micrograms/kg b.w. resulted in an increased activity of the pregnant uterus. These changes lasted 7-8 min and after that time a clear decrease till 40-55 min after the injection was observed. No abortions were observed. In all cases during the post-parturian period, the increase of the electrical activity of the uterine muscular coat was observed. PMID- 2571264 TI - Evaluation of anaesthetic potency of medetomidine-ketamine combination in rats, guinea-pigs and rabbits. AB - In many animal species ketamine hydrochloride is combined with xylazine, an alpha 2-adrenoceptor agonist, to achieve anaesthesia. The purpose of this study was to evaluate the anaesthetic efficacy of various dose combinations of another alpha-2 adrenoceptor agonist medetomidine and ketamine in rats, guinea pigs and rabbits. Both sexes of all three species in groups of five were used. In rats, dose combinations medetomidine/ketamine, both in mg/kg, used were 0.25/60, 1.0/60 and 0.5/70; in guinea pigs 0.5/40 and in rabbits 0.5/25 and 0.5/60. In rats and rabbits the righting reflex was lost within 2-3 min. In rats with doses 0.25/60 and 0.5/75 other reflexes disappeared, reappearing after 20-25 min and 60-70 min in males, the respective values being 120-140 min and 150-180 in females. The righting reflex reappeared after 130-135 and 160-190 min in males and 240 and greater than 300 min in females in the order above. In guinea pigs irrespective of route of ketamine administration all reflexes remained. In rabbits, a dose 0.5/25 caused disappearance of ear pinch reflex. We conclude that the medetomidine-ketamine combination can be used in rats and rabbits as an anaesthetic, and in guinea pigs only for immobilization. PMID- 2571265 TI - Pharmacological effects of medetomidine in humans. AB - Single i.v. doses of medetomidine have been administered to healthy male volunteers in three clinical phase I studies. After an initial open dose-finding study, double-blind, placebocontrolled designs were used. The highest dose tested was 120 micrograms. Medetomidine was well tolerated. It caused dose-dependent decreases of blood pressure (max 22/14 mmHg), heart rate (max 14/min) and cardiac output (max 21%) without subjective sensations of hemodynamic changes or other unexpected side-effects. However, a clear dose-dependent sedative effect, both subjective and objective, and a decrease in salivation was seen after single i.v. doses. Medetomidine had a powerful and dose-dependent effect in reducing noradrenaline (by a maximum of 75%) and increasing human growth hormone levels in plasma. All the observed pharmacological effects are well compatible with an alpha 2-agonistic action of the drug and resemble those seen after i.v. administration of clonidine. However, medetomidine appeared to be more potent and short-acting than clonidine. PMID- 2571266 TI - The clinical efficacy of medetomidine. AB - Studies on the clinical efficacy of medetomidine, a novel alpha-2 adrenoceptor agonist, are reviewed. Medetomidine has been shown to produce a reliable state of sedation, relaxation and recumbency suitable for small animal practice. In dogs, the optimal clinical dose for examinations, clinical procedures and minor surgical interventions seems to be 30-40 micrograms/kg intramusculary and in cats 80-110 micrograms/kg. Other effects of medetomidine reported include bradycardia, nausea and vomiting. Occasional muscle jerkings have been also reported after medetomidine injection. In special investigations, medetomidine has successfully been used in wound suturation and ovariohysterectomy in dogs and for sedation in dogs with heart diseases. Medetomidine-ketamine combination has been shown to be useful for anesthesia and immobilization in cats and zoo animals. The medetomidine-fentanyl combination was tested in dog: The administration of fentanyl increased the sedation and analgesia obtained with medetomidine. Medetomidine appears to be a potent sedative and analgesic agent for clinical use. PMID- 2571267 TI - Sedative and analgesic effects of medetomidine in dogs--an open clinical study. AB - Medetomidine was given parenterally as a sedative and analgesic agent to 283 dogs. Mean doses varied from 17 micrograms/kg for x-ray examinations to 31 micrograms/kg for clinical examinations. An adequate analgesic effect was demonstrated at higher doses than produced sufficient sedation. Potent bradycardia was produced within 15 minutes post administration. No influence of breed, sex or age was found. Vomiting was observed in 8% of the cases. One dog died, showing chronic glomerulonephritis, myocardial necrosis and liver congestion at autopsy. PMID- 2571268 TI - Clinical investigations of medetomidine in dogs. AB - Experiments with 3 doses of medetomidine (20, 40 and 80 micrograms/kg, iv. and im., respectively) were carried out on 90 dogs of 16 breeds in the Small Animal Surgery of the University of Veterinary Science, Budapest. Changes in hematology as well as in AST, AP, BUN, creatinine were studied. Medetomidine administered iv deepened the sedation and lengthened tranquillization dose-dependently. After im administration the sedative effect was still dose-dependent, but the duration of its clinical effectiveness could not be lengthened significantly. The development of the sedation could however, be quickened. The iv administration increased the level of analgesia in proportion to dosage; the im application could change the level of pain-killing effect of the drug, but could not lengthen it. According to the laboratorical determinations, regardless of the dosage and the route of application, medetomidine did not affect the AST and AP enzyme activities, or the BUN and creatinine values. PMID- 2571269 TI - Effects of medetomidine in dogs with mitral regurgitation. AB - Thirteen small breed dogs with heart murmurs due to mitral regurgitation of varying severity received medetomidine as a sedative drug for teeth cleaning procedures. The dose was 30 micrograms/kg as an intramuscular injection. The drug caused a considerable reduction of the body temperatures and the heart rates over the 6 hrs of monitoring. The respiratory rates increased over the first 30 min and later decreased below control values. Systolic arterial blood pressures did not change significantly. Arterial oxygen tensions were significantly lower at 30 min after drug administration, but the arteriovenous oxygen differences and the carbon dioxide tensions changed very little. On electrocardiograms, sinus bradycardia, marked sinus arrhythmia with sinus pauses, AV-blocks 1st. and rarely 2nd. degree, supraventricular premature contractions (4 dogs) and paroxysmal tachycardia (1 dog) were observed. Echocardiograms showed an insignificant reduction of the shortening fraction and an increase of the left ventricular enlargement ratio as well as the left atrial to aortic root ratio at 30 min after drug administration. All measured parameters had returned to normal values (pre drug levels) at 24 hrs after sedation. One dog had delayed cardiovascular complications at 3 hrs after drug administration, (period of prolonged asystole), necessitating the use of the antidote MPV 1248, which successfully reversed the abnormality. Two dogs collapsed briefly, at 4 and 5 hrs respectively, after excitement and struggling against physical restraint. In conclusion, medetomidine will have a safe clinical use for sedation of dogs with murmurs from compensated mitral regurgitation as soon as the antidote MPV 1248 (atipamezole) is licensed for clinical use. This will allow the reversion of the cardiopulmonary effects before the time of expected complications. PMID- 2571270 TI - Suturation of wounds of extremities in dogs using medetomidine as sedative and analgesic drug. AB - The suitability of medetomidine in suturation of wounds of extremities was evaluated. Two doses, 80 micrograms/kg and 40 micrograms/kg, were used. Lidocaine was used only in those cases when the procedure could not be performed without a local anaesthetic. In the group 80 micrograms/kg (22 dogs) the wounds could be sutured in 77% of cases without lidocaine. When the dosage was 40 micrograms/kg (22 dogs), treatment was successful in 55% of cases respectively. It is concluded that medetomidine should be administered at a dosage level of 80 micrograms/kg for analgesia when suturing wounds of the legs of dogs. If the analgesia is not sufficient it can be completed with local infiltration of lidocaine. Medetomidine in combination with lidocaine seems to be very useful for analgesia in minor surgical procedures. PMID- 2571271 TI - The use of medetomidine/fentanyl combinations in dogs. AB - The sedative effects of medetomidine at doses of 20 and 40 micrograms/kg im given alone or followed 16-18 min later by fentanyl (2 micrograms/kg iv) was investigated in 6 bitches of mixed breeds. The higher dose of medetomidine alone caused the greater degree of sedation, but two bitches were only lightly sedated with either dose. Side effects noted in some cases included apparent pain on injection, vomiting on induction of sedation, bradycardia, slowing of respiratory rate, cyanosis and muscular twitching. The intravenous injection of fentanyl caused a marked increase in depth of sedation in all animals, inducing a condition similar to neuroleptanaesthesia in which the eyes were rotated downward and the pedal reflex abolished. Slight twitching and sensitivity to sound occurred immediately after fentanyl injection, but this was transient. The cardiopulmonary effects of medetomidine (40 micrograms/kg im) followed 20 min later by either fentanyl (2 micrograms/kg iv) or a saline placebo were investigated in 4 beagle dogs. Medetomidine caused bradycardia, hypotension and reduced respiratory rate, inducing an intermittent respiratory pattern. The iv injection of fentanyl did not further alter the heart or respiratory rate or blood pressure. However there was a small but significant decrease in arterial oxygen tension and rise in arterial carbon dioxide tension. indicating some respiratory depression. We conclude that the use of intravenous fentanyl to dogs already sedated with medetomidine could prove useful in clinical cases where the initial sedation with medetomidine has proved inadequate. PMID- 2571272 TI - Effects of medetomidine on the experimental auricular pain in dogs. AB - The analgesic effect of medetomidine was studied in ten laboratory beagles. The doses of medetomidine used were 10, 30 and 80 micrograms/kg BW. Xylazine (3 mg/kg BW) was included as a positive control and placebo as a negative control. The test utilized two independently randomized Latin square designs (5 x 5) and was carried out in a double blind fashion. Noxious stimuli were induced with a round pointed thermal probe which was set to produce a temperature of +44.9 +/- 0.2 degrees C. The wall of the external auditory canal was touched with the heated point of the probe. The time period from the start of the attachment to the avoidance reflex was measured and recorded as response time. To avoid tissue damage the heat treatment was stopped after 5 s even though the dog did not respond. Medetomidine clearly prolonged the response time. At the highest dose of medetomidine (80 micrograms/kg) all dogs reached the analgesic stage where they did not respond to the heat treatment. At 30 micrograms/kg of medetomidine half of the dogs had a prolonged response time. The lower dose of 10 micrograms/kg had no appreciable effect. Xylazine-induced analgesia was comparable to that of 30 micrograms/kg of medetomidine: half of the dogs lost their avoidance reflex. In conclusion, medetomidine proved to have a dose dependent analgesic effect in experimental auricular pain in dogs. PMID- 2571273 TI - The use of medetomidine, medetomidine-ketamine combinations and atipamezole at Helsinki Zoo--a review of 240 cases. AB - Medetomidine, medetomidine-ketamine combinations and/or atipamezole were used during 240 immobilizations of 32 non-domestic animal species. Medetomidine ketamine induced reliable immobilization with good physiological quality in a variety of species. Atipamezole was effective in reversing the medetomidine, medetomidine-ketamine or xylazine induced immobilization. PMID- 2571274 TI - Physiological role of alpha 2-adrenoceptors in the regulation of vigilance and pain: effect of medetomidine. AB - The sedative action of alpha 2-adrenergic agonists is generally ascribed to inhibition of the locus coeruleus (LC) and its ascending activating projection to the forebrain, although postsynaptic effects in the neocortex may also play a part. The sedative effect of the drugs varies considerably between different species. Systemic application of alpha 2-adrenergic agents have analgesic effects in addition to their sedative actions. Descending noradrenergic projections from the LC area modulate pain transmission at the spinal level. The purpose of this trial was to investigate the sedative properties of medetomidine in the cat. Medetomidine was injected intramuscularly at dose rates of 0.02, 0.06 and 0.18 mg/kg. Xylazine 3.0 mg/kg and saline were used for comparison. Medetomidine increased drowsy waking at the expense of both arousal and sleep. Nociceptive reflexes were suppressed in the case of the smallest dose of medetomidine, and abolished by the others. The 0.18 mg/kg dose of medetomidine had approximately equal effect on vigilance as 3.0 mg/kg of xylazine. The duration of the effect of medetomidine was dose-dependent. The alpha-receptor blocker phentolamine i.m. reversed the heavy sedation produced by medetomidine. Medetomidine also produced significant bradycardia, but the duration of this effect was not dose-dependent. PMID- 2571275 TI - Pharmacological profiles of medetomidine and its antagonist, atipamezole. AB - Medetomidine, (+/-)-4-[1-(2,3-dimethylphenyl)ethyl]-1H-imidazole, is a very potent, selective and specific full agonist at both pre- and postsynaptic alpha 2 adrenoceptors as demonstrated in several models both in vitro and in vivo. In receptor binding experiments the alpha 2/alpha 1 selectivity ratio of medetomidine is 1620 compared to 260, 220 and 160 for detomidine, clonidine and xylazine, respectively. The alpha 2-adrenoceptor activity of medetomidine resides predominantly in its d-enantiomer (dexmedetomidine). Medetomidine induces a dose dependent decrease in the release and turnover of noradrenaline, dopamine and serotonin in the CNS as measured by changes in metabolite concentrations or using pharmacological intervention techniques. Inhibition of sympathetic tone in the CNS by medetomidine leads for a characteristic pattern of pharmacodynamic responses including e.g. hypotension, bradycardia, sedation, relief of anxiety, analgesia and hypothermia. The potent, dose-dependent sedative effects of medetomidine have been demonstrated in several classical animal models (e.g. decrease in spontaneous motility in rats and mice, potentiation of barbiturate induced anaesthesia in rats and mice, induction of sleep in young chicks). At high doses medetomidine has hypnotic of anaesthetic effects, a property which distinguishes it clearly from detomidine, clonidine and other alpha 2-agonists. The pharmacological, neurochemical and behavioral effects of medetomidine can be inhibited by prior, simultaneous of subsequent administration of a selective and specific alpha 2-antagonist, atipamezole. Besides verifying that the main pharmacodynamic effects of medetomidine are alpha 2-mediated, this finding forms a strong basis for the use of atipamezole as a reversing agent against medetomidine-induced effects in veterinary practice. PMID- 2571277 TI - Pharmacokinetics of medetomidine. AB - The pharmacokinetics of medetomidine administered as a single dose (80 micrograms/kg) were studied in rat, dog and cat with the tritium labelled drug. The results showed a rapid distribution, after an s.c. dose, of medetomidine radioactivity into rat tissues including the brains. In plasma/serum a distribution phase with a half-life of only a few minutes was observed. Peak concentration after i.m. administration (dog & cat) was seen within 0.5 h in complete accordance with the rapid onset of clinical effects. The apparent volumes of distribution ranged from 2.8 (dog, i.v.) to 3.5 l/kg (cat, i.m.) and clearances from 27.5 (dog, i.m.) to 33.4 ml/min kg (dog, i.v.). Elimination of medetomidine from plasma/serum occurred with half-lives ranging from 0.97 to 1.60 h. Differences between dosing routes were small. Elimination of radioactivity from rat brain tissue followed approximately the same time course as elimination from plasma suggesting that termination of clinical effects is controlled by removal of drug from CNS. Excretion of radioactivity was from 28.6 to 74.7% of the dose in three days. In each species most of the activity was excreted in urine. Fecal excretion was significant only in the rat. No measurable levels of the parent drug were found in excreta. Instead a hydroxylated product(s) and (their) conjugates (except in the cat) were present in urine. Other metabolites were not identified. It was concluded that elimination occurs mainly by biotransformation in the liver. PMID- 2571276 TI - Cardiovascular actions of medetomidine and their reversal by atipamezole. AB - The cardiovascular effects of medetomidine were evaluated in conscious spontaneously hypertensive (SH) rats, anaesthetized normotensive rats and cats, and in the cardiovascular system of the pithed rat. The drug proved to be a highly selective alpha 2-adrenoceptor agonist in the cardiovascular system of pithed rats so that both its pre and postsynaptic actions could be antagonized by atipamezole but not by prazosin. Medetomidine showed high stereoselectivity, the pharmacologically active enantiomer being dexmedetomidine. Medetomidine was bradycardic and, after an initial increase in blood pressure, hypotensive in anaesthetized and conscious rats and in anaesthetized cats. The mechanism is activation of alpha 2-adrenoceptors since atipamezole efficiently antagonized the hypotensive and bradycardic action of medetomidine in anaesthetized cats. Finally, it was observed that atropine was not efficient in antagonizing the medetomidine-induced bradycardic response in anaesthetized rats. PMID- 2571278 TI - Dexmedetomidine decreases halothane anesthetic requirements in rats. AB - alpha 2-Adrenergic agonists, such as clonidine, reduce the dose requirements for halothane. Medetomidine is more selective as a full agonist for central alpha 2 adrenoceptors than clonidine and is available as both an active (d) and an inactive (l) isomer. We have used the d-isomer to probe the mediating mechanism for the MAC-sparing effect of this alpha 2-agonist in rats. The dose of halothane which rendered 50% of animals insensitive to a painful stimulus (halothane MAC) was determined in rats (150-200 g) before and after d- or l-medetomidine, 10, 30 and 100 micrograms/kg or saline i.p. To determine whether alpha 2-adrenoreceptors mediated the MAC-sparing effect of dexmedetomidine (d-medetomidine), cohorts of rats (n = 6 for each dose) were pretreated with idazoxan, 10 mg/kg i.p., the highly selective alpha 2-antagonist. To determine whether dexmedetomidine's MAC reducing action was mediated in part through either opiate or adenosine receptors, groups of rats were pretreated with either naltrexone, 5 mg/kg i.p., an opiate antagonist; or 8-phenyltheophylline (8-PT), 2.5 mg/kg i.p., an A1 adenosine antagonist. To determine whether postsynaptic mechanisms mediate the anesthetic-sparing effect of dexmedetomidine, rats were depleted of central norepinephrine stores with 6-OHDA and 4 days later MAC was determined before and after each dose of dexmedetomidine. Dexmedetomidine dose-dependently decreased MAC for halothane such that at the highest dose, halothane could be discontinued for up to 30 min without eliciting a response to tail-clamping. Idazoxan completely prevented the MAC-reducing action of dexmedetomidine, while naltrexone and 8-PT were without effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571279 TI - Pharmacologic characterization of the receptor mediating the hypnotic action of dexmedetomidine. AB - The anesthetic-reducing property of medetomidine far exceeds that seen with other alpha 2-adrenergic agonists (e.g., clonidine). This study examined whether medetomidine possesses hypnotic-anesthetic actions. Dexmedetomidine (the d enantiomer of medetomidine) induced loss of righting reflex in rats (i.e., hypnosis) at doses greater than 100 micrograms/kg i.p.; sleep-time was dose dependent up to 1000 micrograms/kg i.p. The l-enantiomer of medetomidine (MPV 1441) did not induce hypnosis even when administered up to 30,000 micrograms/kg i.p. The centrally-active alpha 2-receptor antagonists, atipamezole (MPV-1248) and idazoxan, dose-dependently decreased the hypnotic action of dexmedetomidine. The peripherally-active alpha 2-receptor antagonist, DG-5128, did not reduce dexmedetomidine-induced hypnosis. The stereospecificity of dexmedetomidine's hypnotic action and its dose-dependent attenuation by alpha 2-antagonists confirmed the involvement of alpha 2-receptors in this effect. The finding that only alpha 2-antagonists with central activity attenuate the hypnotic action of dexmedetomidine, suggests that the mediating alpha 2-receptor is located in the central nervous system. In summary, this study suggests that dexmedetomidine induces hypnosis in rats by activating central alpha 2-adrenergic receptors. This study also suggests that alpha 2-antagonists can be used to reverse the hypnotic effects of dexmedetomidine. PMID- 2571280 TI - The effects of medetomidine, an alpha 2-adrenergic agonist, on ventilatory drive in the dog. AB - alpha 2-Adrenergic agonists have been used as potent adjuncts to anesthesia and have anesthetic properties themselves. For this reason, we studied the effects of medetomidine, and isoflurane (1 MAC) on ventilatory drive in dogs. Six chronically tracheotomized mongrel dogs were studied during spontaneous ventilation. Arterial blood samples were analyzed for pH, PaCO2, and PaO2. Airway O2, CO2, N2, and isoflurane were continuously monitored using a mass spectrometer; respiratory rate was determined. The hypercapnic ventilatory response was assessed using the Read rebreathing technique. Control measurements were made under isoflurane anesthesia. Fifteen minutes after the medetomidine (20 ug/kg) was given and the isoflurane discontinued, all measurements were repeated. Isoflurane levels were 1.38 volume % during the isoflurane test period and had declined to 0.3 volume % by the time the medetomidine measurements were obtained. The slope of the CO2 response curve was significantly steeper after medetomidine (0.582 vs 0.269 1.min-1.mmHg), suggesting less respiratory depression when compared to the measurements under isoflurane. PaCO2 and endtidal CO2 were significantly lower in the medetomidine group. No other significant differences were found. Under these conditions, medetomidine (20 ug/kg) resulted in normal blood gas values with less depression of the hypercapnic response curve than under isoflurane anesthesia. PMID- 2571281 TI - Action of the stereoisomers of medetomidine, in halothane-anesthetized dogs. AB - This study was designed to investigate the anesthetic-reducing and cardiovascular effects of the stereoisomers of medetomidine in halothane-anesthetized male beagles. Anesthesia was induced by mask inhalation of halothane in oxygen, followed by 2h equilibration period. The minimum alveolar anesthetic concentration of halothane which prevented the animals from moving in response to a painful stimulus (MAC of halothane) was determined and baseline hemodynamic function was assessed. Dl-Medetomidine, at 3 doses (1, 3 and 10 micrograms.kg-1) was administered via the right atrial port over 15 minutes while maintaining the dog at its individual MAC for halothane. Ten minutes later, hemodynamic parameters were reassessed. MAC determination was then repeated. In two separate sets of experiments, dogs (n = 5 each) were administered either the d- or the 1 enantiomer of medetomidine and the same procedure followed as described above. MAC for halothane significantly decreased following dl-medetomidine administration in a dose-dependent fashion. The d-isomer displayed similar MAC reducing effects while the 1-isomer was without effect. Neither isomer changed the mean arterial pressure while only and d-isomer significantly decreased heart rate and cardiac output. PMID- 2571282 TI - Effects of acute administration of medetomidine on the behaviour, temperature and turnover rates of brain biogenic amines in rodents and reversal of these effects by atipamezole. AB - The effects of acute administration of medetomidine in rodents were examined. Low doses (2.5 micrograms/kg) were anxiolytic, higher doses (10-100 micrograms/kg) sedating, and above 100 micrograms/kg medetomidine treated rats lost their righting reflex and were hypothermic. These higher doses of medetomidine inhibited the release of noradrenaline (NA), dopamine and serotonin in the central nervous system, inhibition of NA release being the most sensitive to medetomidine. All the above effects could be antagonized by administration of suitable doses of the alpha 2-antagonist, atipamezole, indicating that the actions of medetomidine were mediated via activation of alpha 2-adrenoceptors. PMID- 2571283 TI - Introduction to the clinical pharmacology of medetomidine. AB - Medetomidine is a sedative and analgesic drug intended for use in dogs and cats but it can also be successfully used in many other species. The effect of medetomidine is dose dependent at the recommended dose range (10-80 micrograms/kg for dogs and 50-150 micrograms/kg for cats). At doses higher than the recommended ones the strength of sedation does not increase, only the duration of the effect. From the cardiovascular changes induced with medetomidine, the profound bradycardia is most prominent. It can be transiently prevented with atropine or glycopyrrolate medication. An initial increase in arterial blood pressure followed by a longer lasting slightly hypotensive or normotensive period can be observed. Respiratory frequency tends to decrease but the changes stay within normal limits for resting animals. Vomiting may occur during the induction period of sedation. Occasional muscle jerks can be observed. Hypothermia has been reported in every animal sedated with medetomidine. Medetomidine can be used as preanaesthetic prior to ketamine, barbiturate and halothane anaesthesia. PMID- 2571284 TI - Studies to determine the optimal dose of medetomidine for the dog. AB - This study was designed to determine optimal doses of medetomidine which, when given intravenously or intramuscularly, produce analgesia/anesthesia or tranquility in dogs. Forty-eight mature, mongrel, healthy dogs weighing between 5 and 40 kg were randomized to one of four categories according to the dose of medetomidine given: zero, low, medium, high. Low dose was 280 micrograms/m2* given i.v. or 375 micrograms/m2 given im Medium dose was 750 micrograms/m2 given iv or 1000 micrograms/m2 given im High dose was 1220 micrograms/m2 given iv or 1625 micrograms/m2 given im Dogs were given the compound iv; then at least 2 weeks (100 half-lives) later, it was given i.m. Parameters measuring analgesic/anesthetic effects or ataractic effects were monitored during a control period, and at 1/4, 1/2, 1, 2, 3, 4, 5, and 6 h after dosing. A positive (ideal) response was determined by comparing a score based upon the parameters measured to observations of an ideal response made by the investigators. The numbers of dogs, receiving each dose that manifested ideal responses at 1/2 h and returned to normal enough to be left unattended at 3 h, were determined; and the optimal dose was based upon the lowest dose that produced the desired response and the highest dose that permitted the dogs to be left unattended by 3 h. Doses of medetomidine of 750 micrograms/m2 given i.v. and 1000 micrograms/m2 given i.m. were considered to be optimal of the 3 non-zero doses used.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571285 TI - A comparison of xylazine, acepromazine, meperidine and medetomidine as preanesthetics to halothane anesthesia in dogs. AB - The preanesthetic properties of medetomidine, a novel alpha 2-adrenergic agonist, were compared to the preanesthetic properties of acepromazine, zylazine and meperidine prior to halothane anesthesia. The premedications were given to 23 randomly selected laboratory beagles in doses with sedative effects equal to the 10 micrograms/kg used in the medetomidine group. These dogs also received 0.04 mg/kg of atropine prior to the preanesthetic. A group of 7 dogs with a high medetomidine dose (40 micrograms/kg) was included in the study. Cardiopulmonary and respiratory parameters were recorded at 10 min intervals during surgical stimulus. Dogs receiving 40 micrograms/kg of medetomidine showed bradycardia, but higher blood pressures than dogs in the other groups. Adequate oxygen saturations and perfusion were recorded in all groups. The low 10 micrograms/kg dose of medetomididine had a halothane sparing effect comparable to the other premedications, while the 40 micrograms/kg of medetomidine group showed a clear decrease in halothane consumption. The sedative effect of 10 micrograms/kg of medetomidine was in some instances inadequate for proper manipulation of the dogs, but 40 micrograms/kg produced excessive sedation. Atropine was found to counteract the medetomidine induced bradycardia. PMID- 2571286 TI - Aortic arch syndrome--Takayasu's arteritis in Nigeria. AB - Four cases of aortic arch syndrome (Takayasu's arteritis), all in Nigerian females, are presented. Our clinical and roentgenological findings are described and the disease briefly discussed. PMID- 2571287 TI - Nonsustained ventricular tachycardia arising from the right ventricular outflow tract. AB - Characteristics of left bundle branch block morphology, inferiorly directed frontal plane QRS axis and repetitive nonsustained salvos were used to define a discrete subgroup of patients with ventricular tachycardia (VT). The origin of this tachycardia was thought to be the right ventricular outflow tract. Twenty six patients with this definition (group 1) were compared with 29 consecutive patients with all other forms of VT (group 2). When compared with patients in group 2, group 1 patients were younger (average age 37 vs 51 years, p less than 0.005), had less structural heart disease (2 of 26 vs 25 of 29 patients, p less than 0.005) and had a better prognosis (no deaths) after an average follow-up time of 28 months in comparison with 5 deaths after an average follow-up of 35 months (p less than 0.05). Induction of VT was possible using isoproterenol infusion in 14 of 20 group 1 patients, but no VT could be induced in 9 group 2 patients (p less than 0.05). Exercise stress testing induced VT in 11 of 21 group 1 patients and 2 of 9 group 2 patients (p greater than 0.05). Programmed electrical stimulation failed to induce VT in 9 group 1 patients, but did induce it in 15 of 20 group 2 patients (p less than 0.005). Successful therapy in group 1 patients was achieved by beta blockers alone (7 patients), beta blockers plus type 1A antiarrhythmic drugs (9 patients), procainamide alone (2 patients), sotalol (3 patients) and amiodarone (2 patients). Three patients were not treated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571288 TI - Influence of beta blockade on exercise capacity and heart rate response after human orthotopic and heterotopic cardiac transplantation. AB - It has been reported that use of beta blockers may not be safe after cardiac transplantation because the denervated hearts may be largely dependent on circulating catecholamines to increase cardiac output. Therefore, the effects of intravenous propranolol were studied during maximal treadmill exercise in 7 patients with heterotopic and 6 with orthotopic cardiac transplantations. An average decrease of about 15% in exercise duration (p less than 0.001), a 34% reduction in systolic blood pressure increase (p less than 0.05) and a 40% attenuation in heart rate increase (p less than 0.001) were observed after beta blockade. In patients with heterotopic transplantation, beta blockade produced similar effects on heart rate in the denervated donor hearts and the innervated recipient hearts during and after mild exercise. During peak exercise, beta blockade attenuated the rate to a greater extent in the donor hearts. Although the denervated donor heart is more sensitive to beta blockade than the innervated recipient heart during exercise, no adverse effects were observed. Beta-blocker therapy should be considered for cardiac transplant patients if longer-term studies confirm their safe use in these patients. PMID- 2571289 TI - Nifedipine and systemic hypertension. AB - Nifedipine reduces blood pressure predominantly by reducing systemic vascular resistance due to a direct vasodilating action on the arterioles. This peripheral vasodilation appears greater the more severe the hypertension. Studies have demonstrated an additive effect of beta blockers and converting-enzyme inhibitors in patients not controlled with nifedipine alone. Although there is a controversy about whether diuretics have an additive effect on blood pressure in patients already taking nifedipine, it would appear that the blood pressure-lowering effect of thiazide is blunted. Studies have shown that a high sodium intake may enhance the acute blood pressure-lowering effect of nifedipine. Nifedipine does cause a long-term reduction in sodium balance. PMID- 2571290 TI - New concepts on the central regulation of blood pressure. Alpha 2-adrenoceptors and "imidazoline receptors". AB - The most usual hypothesis to explain the central hypotensive effect of clonidine like substances was to admit that these drugs stimulated alpha 2-adrenoceptors within the brainstem. Now it has been demonstrated that neither the endogenous ligand to the alpha-adrenoceptors, noradrenaline, nor any other catecholamine or phenylethylamine was hypotensive in the medullary nucleus reticularis lateralis, where all imidazolines proved to be such. Recently, a membrane receptor population sensitive to clonidine and insensitive to catecholamines was described within the nucleus reticularis lateralis; this subgroup of receptors represented 20 to 30 percent of the [3H]clonidine binding sites in the bovine nucleus reticularis lateralis and 100 percent within the human nucleus reticularis lateralis region. Thus, the existence of such imidazoline specific receptors was clearly established and the endogenous ligand for those receptors, which is neither a catecholamine nor likely a peptide, is under processing for purification. Therefore, it appeared that the hypotensive effect of substances with an imidazoline or imidazoline-like structure might be due to their action within medullary receptors specific for this endogenous ligand temporarily named "clonidine displacing substance." Rilmenidine, structurally close to imidazolines, also interfered with these receptors. The central component of its hypotensive effect was recently confirmed in rabbits, where its central cardiovascular effects were antagonized by "the clonidine displacing substance." Although exhibiting a lower affinity than the reference substance for these receptors, rilmenidine might have a higher selectivity, thus explaining its restricted side effects. A structure-activity study with this molecule would bring a confirmation to these first observations. PMID- 2571291 TI - Recent advances in the pharmacology of rilmenidine. AB - The antihypertensive properties of rilmenidine, an oxazoline derivative, have been demonstrated in several experimental models of hypertension after short- or long-term administration. In pentobarbitone-anesthetized spontaneously hypertensive rats, intravenous rilmenidine (0.1 to 1 mg/kg) dose-dependently reduced blood pressure and heart rate. Upon long-term subcutaneous infusion (5 to 15 mg/kg per day) in conscious spontaneously hypertensive rats, rilmenidine induced a dose-dependent decrease in both cardiovascular parameters. In conscious sino-aortic denervated dogs, rilmenidine (1 mg/kg orally for two weeks) significantly reduced blood pressure and heart rate. The hypotensive action of rilmenidine is mediated through a reduction in peripheral sympathetic tone, resulting from a central action and possibly a peripheral action. Rilmenidine also decreases catecholamine release from the adrenal medulla which might contribute to the antihypertensive effect. Therefore, rilmenidine acts similarly to clonidine and related compounds in order to lower blood pressure, i.e., reduction of sympathetic tone. Nevertheless, although it binds to alpha 2 adrenoceptors, rilmenidine did not cause sedation in animal models: at doses up to 10 mg/kg in mice and rats, it did not prolong the barbiturate-induced sleeping time and did not modify the spontaneous locomotor activity in rats at doses up to 2.5 mg/kg. These results demonstrate a dissociation between sedative and antihypertensive effects of rilmenidine. Three hypotheses have been proposed to explain why this drug is almost devoid of sedative activity in animal experimental models: (1) unknown properties counteracting the alpha 2 adrenoceptor-mediated sedation; (2) a preferential action at the peripheral level; (3) central receptors involved in sedation and hypotension may be different. The intimate mechanism underlying the hypotensive effects of rilmenidine is currently under investigation. The evidence for rilmenidine binding on central sites named "imidazoline sites" involved in blood pressure regulation could possibly provide further insight into its mechanism of action and explain the duality of its effects. PMID- 2571292 TI - Rilmenidine: a novel antihypertensive agent. AB - Rilmenidine is a novel antihypertensive agent related to alpha 2-agonists. Pharmacologic and clinical data concerning efficacy and acceptability of rilmenidine, its effects on physiologic functions, and hypothesis of mechanism of action are described in this report. The dominant aspect of rilmenidine is the observed dissociation at therapeutic doses between antihypertensive and central effects. In this respect, several findings resulting from fundamental pharmacology, clinical pharmacology, and therapeutics differentiate rilmenidine from clonidine and related drugs. Hypotheses to explain the results observed with rilmenidine involve unknown properties that could counteract with the alpha 2 adrenoceptor-mediated sedation, respective contribution of central and peripheral components to the activity of rilmenidine, and possible contribution of imidazoline receptors. PMID- 2571294 TI - Investigation of alpha 2-adrenoceptors in humans. AB - Partial alpha 2-agonists are thought to lower blood pressure principally by stimulation of postsynaptic central nervous system alpha 2-adrenoceptors. It is possible for some to have also a peripheral action, either by acting on the inhibitory presynaptic or as an antagonist at the postsynaptic alpha 2 adrenoceptor. In order to assess these other actions, indices are required for these receptors. Plasma growth hormone is used for the central nervous system and skin blood flow for peripheral postsynaptic alpha 2-adrenoceptors. Plasma insulin is also an index for the latter, but a double-site assay is required to detect decreases within the fasting range. Plasma noradrenaline reflects both central nervous system and peripheral modulation of sympathetic nerve activity, so that "dynamic" tests (described next) are required to dissect these two control mechanisms. Selective peripheral alpha 2-adrenoceptor stimulation or blockade may be attractive for various therapeutic applications such as hypertension, diabetes, and Raynaud's phenomenon. The balance of opposing pre- and postsynaptic alpha 2-adrenoceptor effects will be important and this may require more dynamic tests than already mentioned, such as comparing the effect of alpha methylnoradrenaline on plasma noradrenaline (presynaptic effect) and skin blood flow (postsynaptic effect) in the presence and absence of the partial alpha 2 agonist or antagonist under investigation. Since alpha-methylnoradrenaline does not penetrate the central nervous system, any blockade of its action by a drug must be peripherally mediated. Examples of all these indices, investigations, and drugs are presented. PMID- 2571293 TI - Bronchial effects of alpha 2-adrenoceptor agonists and of other antihypertensive agents in asthma. AB - The respective prevalence of hypertension and asthma is sufficient for their combined existence to be far from rare. The effects of certain antihypertensive drugs, e.g., alpha 2-adrenoceptor agonists, on the bronchi may be either harmful or beneficial. When inhaled, alpha 2-agonists reduce the immediate bronchial response to allergens, whereas when ingested they aggravate the bronchial response to histamine and all the more so when their effect on the central nervous system is greater. Therefore, there has been much interest in agents such as the new oxazoline derivative, rilmenidine, which has less central effects than clonidine, an imidazoline compound of reference. Calcium antagonists inhibit smooth muscle contraction and release of mast cell inflammatory mediators. In asthmatic subjects, their short-term administration leads to a modest improvement in spontaneous bronchial obstruction, has only a partial protective action against various nonspecific or allergenic stimuli, and slightly reinforces the beneficial effect of beta 2-agonists. Beta-adrenoceptor antagonists aggravate bronchial obstruction and nonspecific bronchial hyperreactivity in asthmatic subjects. These harmful effects are dose-dependent, have even been reported after the administration of eyedrops, and are common to all beta-blockers. Angiotensin converting enzyme inhibitors increase bronchial hyperreactivity in patients who develop cough during treatment and may, in certain cases, worsen or even induce asthma, probably by opposing inactivation by hydrolysis of tachykinins and of bradykinins. PMID- 2571295 TI - Renal excretory actions of antihypertensive agents. Effects of rilmenidine. AB - The acute renal excretory actions of several antihypertensive agents were assessed in healthy volunteers. Rilmenidine (1 mg) did not affect water and electrolyte balance. Beta-adrenergic blockers had little effect upon urinary composition, whereas diuretics caused significant urinary losses of fluid, sodium, potassium, and magnesium as expected. The vasodilators ketanserin and flosequinan did not affect water and electrolyte excretion. Ketanserin increased urate output in common with angiotensin-converting enzyme inhibitors, which may act as facultative diuretics. Combining the angiotensin-converting enzyme inhibitor captopril (100 mg) or rilmenidine (1 mg) with hydrochlorothiazide (25 mg) had little influence upon the known actions of the diuretic. When the beta blocker pindolol (10 mg) was given with the diuretic clopamide (5 mg), beneficial effects were noted; potassium and magnesium losses were markedly reduced, whereas the natriuretic and diuretic effects of clopamide were unchanged. These short term studies in healthy volunteers provide useful pointers to some possible pharmacodynamic effects of antihypertensive drugs in patients. PMID- 2571296 TI - Rilmenidine and vigilance. Review of clinical studies. AB - In this study, the possible effects of rilmenidine on vigilance are evaluated. Sedation is the most disturbing side effect of alpha 2-agonists, especially during the first weeks of treatment. The level of vigilance was first determined by assessing drowsiness using visual analogue scales and/or by several psychometric tests in four pharmacoclinical studies in healthy subjects or in hypertensive patients: three studies with single administration of rilmenidine (0.5 to 3.0 mg) and one study with repeated administration for three days. These studies were double-blind, Latin-square designed, and controlled versus placebo (in all studies) and versus clonidine (in three studies). Analysis of these results illustrated that after short-term and repeated administration: (1) the effects on vigilance observed with rilmenidine 1 mg did not differ statistically from data observed with placebo; and (2) sedative effects observed with clonidine were significantly greater than with rilmenidine, at equihypotensive doses. Daytime drowsiness was systematically assessed and graded by inciting questioning at each visit in five clinical studies. Ambulatory hypertensive patients were treated with rilmenidine (1 mg per day or 1 mg twice a day). These studies were controlled versus placebo (one study for two weeks, 120 patients; and one study for one month, 126 patients), hydrochlorothiazide (six weeks, 56 patients), clonidine (six weeks, 333 patients), and methyldopa (three months, 157 patients). The results showed that: (1) drowsiness observed with rilmenidine did not differ statistically from that observed with placebo or diuretic; and (2) drowsiness occurred less frequently with rilmenidine than with reference alpha 2-agonists at equihypotensive doses. In conclusion, these results confirm in current clinical use the dissociation already observed in laboratory animals between the antihypertensive effects and the sedative effects and may distinguish rilmenidine among alpha 2-agonists. PMID- 2571297 TI - Role of dopaminergic and adrenergic receptors in the pathogenesis of arterial lesions induced by fenoldopam mesylate and dopamine in the rat. AB - Fenoldopam mesylate (FM), a selective, postjunctional, dopaminergic (DA1) vasodilator, causes a novel lesion of large caliber splanchnic arteries (100 to 800 microns) in the rat characterized by necrosis of medial smooth muscle cells and hemorrhage. FM does not induce lesions in other vascular beds of the rat or in dogs or monkeys. Dopamine, like FM, causes hemorrhagic lesions of large caliber splanchnic arteries in the rat, as well as fibrinoid necrosis of small caliber arteries (less than 100 microns) of the splanchnic, cerebral, coronary, and renal vascular beds. Dopamine, an alpha- and beta-adrenoceptor and dopaminergic agonist, is used clinically, principally as a pressor agent. Because these arterial lesions were believed to result from the pharmacologic activity of these two compounds, the role of vascular receptor subtypes in their pathogenesis was investigated. Rats were coexposed to either FM or dopamine and a variety of receptor antagonists (alpha, beta, DA1, DA2, and 5HT2). In rats coexposed to the alpha-adrenoreceptor antagonist phenoxybenzamine (PBZ) and either FM or dopamine, the incidence and severity of hemorrhagic lesions of large caliber arteries were increased; PBZ, however, prevented the formation of dopamine-induced fibrinoid lesions in arteries of small caliber. SK&F 83566-C, a selective DA1 dopaminergic receptor antagonist, prevented the induction of FM and dopamine-induced hemorrhagic lesions of large caliber arteries. Rats exposed concurrently to dopamine, phenoxybenzamine, and SK&F 83566-C were free of all arterial lesions. The other receptor antagonists tested did not prevent arterial injury. Thus, the induction of splanchnic arterial lesions in the rat by dopamine and FM is caused by stimulation of, and interaction between, alpha-adrenoceptors and dopaminergic DA1 receptors. Activation of the postjunctional, dopaminergic (DA1) receptor is causally associated with the induction of novel hemorrhagic lesions of large caliber splanchnic arteries in the rat. PMID- 2571298 TI - Release of immunoreactive angiotensin II from neuronal cultures: adrenergic influences. AB - The effects of adrenergic drugs on the release of immunoreactive angiotensin II (ANG II-ir) from brain cells in culture were examined. In neuronal cultures, basal release of Ang II-ir was 43.65 +/- 7.44 pg/5-min incubation period (n = 14 experiments; 52 individual determinations), and in astrocytic glial cultures, it was 21.76 +/- 5.7 pg (n = 8 experiments; 24 individual determinations) when cells were exposed to buffer alone. Incubation of neuronal cultures with the alpha 2 adrenergic antagonist yohimbine (0.1-50 microM, 5 min) caused concentration dependent increases in ANG II-ir release above basal levels. Analysis of the released material by high-pressure liquid chromatography revealed that authentic ANG II was present. No increase in the release of ANG II-ir was seen from glial cells. Experiments using neuronal cultures revealed that the yohimbine-induced release of ANG II-ir may be secondary to increased norepinephrine (NE) release. Incubation of neuronal cultures with NE (10 nM-50 microM) caused concentration dependent increases in the release of ANG II-ir. This effect of NE was not inhibited by the alpha 1-adrenergic blocker prazosin. However, a weaker release of ANG II-ir from neuronal cultures was stimulated by the beta-adrenergic agonist isoproterenol at 100 microM. These data show that ANG II-ir can be released from neuronal but not glial cell cultures by adrenergic receptor-mediated mechanisms. PMID- 2571299 TI - Atrial natriuretic factor suppresses neural stimulation of renin release in dogs. AB - The effects of atrial natriuretic factor (ANF) on neural control of renin release and sodium excretion by the kidney were examined in pentobarbital-anesthetized dogs. Electrical stimulation of the renal nerves (RNS, 1 Hz) increased the renal secretion rate of renin (RSR) by 627 +/- 141 ng angiotensin I (ANG I)/min and that of norepinephrine (NESR) by 22.2 +/- 5.9 ng/min. Furthermore, urinary sodium excretion (UNaV) was decreased by 59 +/- 7%, with little change in either renal blood flow (RBF) or glomerular filtration rate (GFR). Intrarenal arterial infusion of ANF (alpha-human atrial natriuretic peptide; 10 ng.kg-1.min-1) increased basal UNaV about twofold but had no effect on basal RBF or GFR. The RNS induced increase in RSR during ANF infusion (198 +/- 117 ng ANG I/min) was significantly lower than that before the infusion (P less than 0.05), whereas the RNS-induced changes in NESR (27.1 +/- 8.5 ng/min) and UNaV (51 +/- 11%) were unaffected. These results suggest that neural stimulation of renin release, but not of tubular sodium reabsorption, can be suppressed by exogenously administered ANF at a dose that does not affect glomerular filtration or renal neurotransmitter release. PMID- 2571300 TI - Alpha-adrenoceptors and insulin release from pancreatic islets of normal and diabetic rats. AB - The role of alpha-adrenoceptors in the regulation of glucose-induced insulin release (GIR) was investigated in islets of normal and neonatally streptozotocin injected non-insulin-dependent diabetic rats (STZ). In normal islets GIR was suppressed to approximately 50% by 10(-8) M of the alpha 2-adrenergic agonist UK 14304, whereas 10(-9) M of the agonist induced a similar inhibition in STZ islets. In normal islets, suppression of GIR by UK 14304 (10(-8) M) was totally antagonized by 10(6) M idazoxan (alpha 2-antagonist) or 10(6) M phentolamine (alpha 1 + alpha 2-antagonist). In STZ islets, the inhibitory effect of UK 14304 (10(-9) M) was entirely reversed by 10(-5) M idazoxan or 10(-6) M phentolamine. The alpha 1-antagonist prazosin (10(-7)-10(-5) M) was without effect on insulin release suppressed by UK 14304 in normal and STZ islets. Insulin release at 3.3, 8.3, or 16.7 mM glucose was augmented by phentolamine but not by idazoxan. It is concluded that the inhibitory effect of catecholamines on insulin release is mediated by alpha 2-receptors in normal and STZ islets. Phentolamine augments basal and glucose-induced insulin release by a mechanism that does not involve alpha 2-adrenoceptors. PMID- 2571301 TI - VIP and acetylcholine: neurotransmitters in esophageal circular smooth muscle. AB - Vasoactive intestinal polypeptide (VIP) and acetylcholine were evaluated as possible inhibitory and excitatory neurotransmitters in the cat esophageal circular smooth muscle. Circular muscle strips 2 mm in thickness were obtained from 1 to 3.4 cm above the lower esophageal sphincter and tested in vitro. Muscle strips contracted with bethanechol (10(-5) M) were relaxed by electrical stimulation (0.5-5 Hz) and by VIP (10(-8)-10(-6) M). Relaxation induced by electrical stimulation was blocked by tetrodotoxin, whereas VIP-induced relaxation was not affected. Highly specific VIP antiserum (5%) antagonized both VIP and electrically induced relaxation, and the antagonism was eliminated when the antiserum was neutralized with VIP (10(-6.5) M). Dopamine (10(-4) M) reduced the relaxation induced both by exogenous VIP and by electrical stimulation but did not affect the relaxation caused by sodium nitroprusside (10(-8)-10(-5) M). In untreated strips, physostigmine (10(-10)-10(-8) M) enhanced the off contraction in response to electrical stimulation, whereas atropine caused a dose dependent reduction with complete abolition at 10(-4) M. These data suggest that in the esophagus inhibition and excitation are mediated by distinct mechanisms: VIP mediates inhibition and acetylcholine is responsible for the off contraction in response to electrical stimulation. PMID- 2571302 TI - The dopamine receptor in adult and maturing kidney. AB - Dopamine, like other neurotransmitters, exerts its biological effects by occupation of specific receptor subtypes. The dopamine receptors in the central nervous system and certain endocrine organs are classified into the D1/D2 subtypes. Outside the central nervous system, the dopamine receptors are classified into the DA1/DA2 subtypes. The D1/D2 and DA1/DA2 receptor have marked similarities and some differences, the most notable of which is the lower affinity of the DA dopamine compared with the D dopamine receptor. DA1 receptor activation increases renal blood flow (RBF); stimulation of DA1 and DA2 receptors may also increase glomerular filtration rate (GFR). DA1 agonists inhibit fluid and electrolyte transport indirectly via hemodynamic mechanisms and directly by occupation of DA1 receptors in specific nephron segments. In the proximal tubule, DA1 agonists simulate adenylate cyclase and inhibit Na+-H+ antiport activity. They also increase phospholipase C and inhibit Na+-K+-ATPase activity (presumably as a consequence of protein kinase C activation). The latter effects may be facilitated by DA2 agonists. In cortical collecting ducts, dopamine antagonizes the effects of mineralocorticoids and the hydrosomotic effect of antidiuretic hormone. It has also been suggested that DA1 may also decrease sodium transport by influencing other hormones, such as atrial natriuretic peptide. Studies of dopamine in the young are complicated because of the propensity for dopamine to stimulate alpha-adrenoceptors. Dopamine alone may actually decrease RBF in the perinatal period. In some animals, the renal vasodilatory and natriuretic effects of dopamine increase with age. Renal tubular DA1-stimulated adenylate cyclase activity increases, whereas renal tubular DA1 receptors decrease with age. Renal DA2 receptor density is greater in the fetus; after birth renal DA2 receptors do not change. Endogenous dopamine may regulate sodium excretion in the young differently than in the adult. In the adult, sodium surfeit is associated with an increase in urinary dopamine; the opposite occurs in the young. A decrease in dopamine production or blockade of dopamine receptors results in an antinatriuresis in the adult; dopamine blockade in the young results in a natriuresis. It remains to be determined whether these age-related differences in dopamine effects are due to changes in receptor DA subtype density, second messengers, and/or interaction with other receptors. PMID- 2571303 TI - Central effects of somatostatin: pressor response, AVP release, and sympathoinhibition. AB - The effects of intracerebroventricular (icv) administration of somatostatin(1-14) (SS1-14) on mean arterial blood pressure (MAP), heart rate (HR), plasma arginine vasopressin (AVP) concentration, and splanchnic nerve activity (SpNA) were studied in conscious rats. In addition, the effects of peripheral alpha adrenergic receptor blockade with prazosin, vasopressinergic V1-receptor blockade with [d(CH2)5Tyr(Me)]AVP, and chronic bilateral sinoaortic denervation (SAD) on central SS1-14-induced MAP, HR, and SpNA responses were investigated. SS1-14 icv elicited dose-dependent increases in MAP and plasma AVP concentration as well as decreases in HR and SpNA. Prazosin iv did not significantly affect SS1-14-induced pressor and bradycardic responses but augmented the decrease in SpNA. The V1-AVP receptor antagonist iv significantly attenuated the effects of SS1-14 icv on MAP, HR, and SpNA. Following SAD the pressor responses to SS1-14 icv were significantly enhanced and were associated with significantly smaller decreases in HR and SpNA. We conclude that central administration of SS1-14 causes a pressor response via release of AVP while at the same time inhibiting peripheral sympathetic outflow. Our results support the hypothesis that SS1-14 in the brain by its effects on AVP release and sympathetic outflow may participate in central cardiovascular regulation. PMID- 2571305 TI - Time course of antipsychotic effects of neuroleptic drugs. AB - The authors reviewed relevant studies in an attempt to define the onset and time course of antipsychotic effects of neuroleptic drugs. After excluding open trials, studies of chronically psychotic patients, and studies not using a placebo or nonneuroleptic sedative as a control, they found only five reports suitable for analysis. Among these, the degree of patients' improvement during neuroleptic treatment was similar regardless of the duration of the study. Also, in studies comparing neuroleptics with sedatives, similar improvement was observed with both treatments. Although neuroleptic drugs have been used clinically for 35 years, the timing of their specific therapeutic effects remains unclear. PMID- 2571306 TI - Benzodiazepine use and abuse by patients at outpatient clinics. AB - The charts of 2,719 patients from several outpatient clinics were reviewed for evidence of use and abuse of benzodiazepines. According to the chart data and interviews with physicians, no patient met the criteria for benzodiazepine abuse or dependence. PMID- 2571304 TI - Second messenger systems and psychoactive drug action: focus on the phosphoinositide system and lithium. AB - Most models of psychotropic drug action have emphasized effects on the release or uptake of neurotransmitters as well as interactions with receptors. Stimulation of neurotransmitter receptors triggers a complex array of electrical and biochemical actions. Recent experimental advances have greatly clarified the second messenger systems underlying neurotransmitter actions, suggesting novel sites of action for psychotropic agents. The authors emphasize the phosphoinositide system and the implications of its involvement for the therapeutic actions of lithium. PMID- 2571307 TI - A clinical study into the possible intrinsic bradycardic activity of vecuronium. AB - Forty female patients received a standardised anaesthetic technique with thiopentone sodium and enflurane. Half of the patients received, under double blind conditions, either physiological saline or glycopyrronium before induction of anaesthesia; 10 minutes after induction of anaesthesia, all patients received vecuronium 0.1 mg/kg. A further 10 patients received neither glycopyrronium nor vecuronium. The results show that vecuronium per se does not produce a decrease in heart rate. PMID- 2571308 TI - Temazepam or midazolam for night sedation. A double-blind study. AB - The effects of oral temazepam (20 mg), oral midazolam (15 mg) and a placebo were compared for night sedation on the evening prior to surgery in a double-blind study. Patients in the placebo group had significantly worse sleep than those in the temazepam (p = 0.004) or midazolam groups (p = 0.04). There was no significant difference between the two drug groups, nor between the residual effects of the three treatments. Temazepam appears to be somewhat more effective than the ultrashort-acting midazolam in pre-operative transient insomnia. PMID- 2571309 TI - Weight determined dosage of vecuronium bromide. PMID- 2571310 TI - Quantitative electron microscopic immunocytochemistry of neuroactive amino acids. AB - Amino acids are of crucial importance in brain function, not only as metabolic intermediates and building blocks of proteins, but also as mediators of interneuronal communication. This dual role of the amino acids distinguishes them from other neurotransmitter candidates, and implies that they are unlikely to be restricted to neurons using them as transmitters. This calls for a quantitative approach when attempts are made to analyse the distribution of transmitter amino acids by means of immunocytochemistry. The present review deals with recent methodological developments that have made it possible to utilize specific antisera to explore the cellular and subcellular distribution of neuroactive amino acids in a quantitative manner. PMID- 2571311 TI - Ultrastructural study of the endocrine cells of the gut of Testudo graeca (Chelonia). AB - The digestive tract of Testudo graeca (Chelonia) was investigated by means of electron microscopy using both conventional and immunocytochemical techniques. EC , L-, D-, G-, B-, N- and EC-L-cells were detected. These cells share several common ultrastructural characteristics with the endocrine cells of mammals (i.e. clear cytoplasm, prominent Golgi apparatus, secretory granules etc.). EC and D1 cells have so far not been described in the esophagus of any animal species; in the present study these cells have been observed in the esophagus of T. graeca. Of special interest was the presence of B-cells in the intestine, suggesting that the migration of B-cells from the gut to the pancreas to constitute pancreatic islets is not concluded in T. graeca. The present study demonstrates that the gut endocrine system of T. graeca is a complex structure containing a large variety of endocrine cell types similar in morphology to those found in higher vertebrates. PMID- 2571312 TI - The distribution of zinc in the forebrain and midbrain of the lizard Gekko gecko. A histochemical study. AB - The distribution of zinc in the forebrain and midbrain of the lizard Gekko gecko was studied with the recently modified Timm method. Areas with a high intensity of staining are almost exclusively found in the telencephalon, although also some structures in the diencephalon display notable staining. Cortical areas that stain heavily are the deep zone of the subcortical layer of the small-celled medial cortex, the longitudinal association bundle that encompasses the large celled medial cortex, and the dorsal cortex. Of the subcortical areas, particularly the anterior septal nucleus shows a high intensity of staining. Moderate to dense Timm staining is further observed in the ventral part of the anterior lateral cortex, the lateral septal nucleus, the striatum, the amygdaloid complex, and the dorsal ventricular ridge. Staining in the diencephalon is primarily confined to the stria terminalis and the ventromedial hypothalamic nucleus, whereas in the midbrain weak staining is observed in the ventral tegmental area and the periventricular layers of the tectum and the tegmentum. The presence of zinc in the gekkonid brain is discussed in relation to connections and neurotransmitters as studied in same species. Moreover, similarities in pattern of staining for zinc in mammals and reptiles and possible evolutionary implications are mentioned. PMID- 2571313 TI - Sperm nuclear chromatin heterogeneity in infertile subjects. AB - Sperm chromatin heterogeneity has been evaluated in infertile males affected by different testicular diseases: 37 subjects had undergone orchidopexy in childhood (ex-cryptorchid), 50 were affected by idiopathic varicocele, 18 had a history of bilateral post-parotitis orchitis and 23 were "idiopathic infertiles". All subjects, except post-parotitis orchitic patients, exhibited significantly higher sperm chromatin heterogeneity than controls, with the highest incidence in ex cryptorchid and in idiopathic infertiles. Ex-cryptorchid subjects also presented a significant positive linear correlation (p less than 0.001) between degree of sperm chromatin abnormality and percentage of morphological sperm alterations. Four monolateral ex-cryptorchid subjects showed a higher percentage of chromatin heterogeneity even when the cryptorchid testis had been removed during orchidopexy. In patients affected by varicocele, we also observed a significant correlation between chronological age and percentage of chromatin alterations. The results are discussed in relation to the pathogenesis of the disease concerned. Since sperm chromatin heterogeneity appears to be strongly involved in the development of infertility, we would suggest that it should be evaluated in routine diagnostic procedures of male infertility. PMID- 2571314 TI - Clinical effect of cryptocur by undescended testicles. AB - As study of the clinical effectiveness of Cryptocur nasal spray (Hoechst) containing LH-RH in 46 cases of maldescended testicles of 40 children is presented by the authors. They concluded that its application has several advantages. Few side effects occur and the treatment is well tolerated by the patients. So the clinical use of Cryptocur is suggested. PMID- 2571315 TI - Increased requirements for continuously infused vecuronium in critically ill patients. PMID- 2571316 TI - Atrioventricular Mobitz I block during propofol anesthesia for laparoscopic tubal ligation. PMID- 2571317 TI - [Evaluation of enzymuria as a tracer in nephrotoxicity: results of a multicenter study]. AB - Urinary excretion of three enzymes of different subcellular location in kidney tissue, alanine aminopeptidase (AAP), gammaglutamyltransferase (GGT), N acetyl beta-D glucosaminidase (NAG), was carried out in 79 healthy adults and 108 healthy children and in 69 adults with various therapies: antibiotics (32 cases), non steroidal anti-inflammatory drugs (NSAIDs) (22 cases), cisplatinum (12 cases) and cyclosporine (3 cases). A circadian rhythm has been shown in children. In patients treated with antibiotics, the importance and duration of the increased enzymes urinary excretion were variable but the excretion of AAP was always higher than that of GGT and NAG. Short term therapies by NSAIDs were without influence on enzymuria but long term therapies produced a moderate increase of NAG excretion. Enzymuria increased immediately after cisplatinum administration and decreased after each daily dose, except in patients with previously high creatininemia. Cyclosporine induced a slight increase in AAP and NAG excretion. Enzymuria, thus, increased early reflecting a toxic effect of the drug at the cellular level whereas creatininemia increase, marker of renal fonctionnal insufficiency, occurs only occasionally and lately. PMID- 2571318 TI - Long-term complete chimerism and stable hematopoiesis in beagles after fetal liver hematopoietic stem cell transplantation. AB - Karyotype analysis of bone marrow cells was performed from 6 Beagles that had received sex-mismatched fetal liver hematopoietic stem cell grafts 2.4 to 6.5 years earlier. Two dogs received dog leukocyte antigen-matched fetal liver cells and 4 received dog leukocyte antigen-mismatched cells. In each dog, all 25 evaluated metaphase spreads were of donor genotype. All dogs had normal hemograms and repopulation of bone marrow and no dog developed graft-vs-host disease. PMID- 2571319 TI - Evidence for the involvement of alpha 2-adrenoceptors in the emetic action of xylazine in cats. AB - Intramuscular injection of xylazine induced dose-dependent vomiting in cats (ED50 = 0.277 mg/kg); administration of standard dose of xylazine (2 mg/kg, 2 times the 100% emetic dose) induced vomiting in 100% of the cats studied. The xylazine induced vomiting was antagonized by adrenoceptor antagonists possessing alpha 2 blocking activity, which were yohimbine, tolazoline, and phentolamine. Of these antagonists, yohimbine was the most effective; the maximal antagonistic effect was seen at 1 mg of yohimbine/kg, a dose at which the other drugs had little or no effect. At the doses studied, prazosin and phenoxybenzamine, adrenoceptor antagonists with alpha 1-blocking activity, did not prevent vomiting induced by xylazine. Beta-Adrenoceptor (propranolol), dopamine receptor (domperidone and chlorpromazine), a cholinoceptor (atropine), an opiate receptor (naloxone), and a histamine-receptor (diphenhydramine) antagonists, at the doses studied, did not prevent xylazine-induced vomiting. Pretreatment with 6-hydroxydopamine failed to prevent xylazine-induced vomiting. These results indicated that xylazine-induced vomiting in cats is mediated by alpha 2-adrenoceptors and suggested that the alpha 2-adrenoceptors mediating the vomiting attributable to xylazine may not be presynaptic alpha 2-receptors located on noradrenergic nerve terminals. PMID- 2571320 TI - Inhibition of hypertonic saline-induced bronchoconstriction by terfenadine and flurbiprofen. Evidence for the predominant role of histamine. AB - We investigated the possible inhibitory effects of terfenadine, a histamine H1 receptor antagonist, and flurbiprofen, a cyclooxygenase inhibitor, on the bronchoconstrictor effect of inhaled 3.6% hypertonic saline in a randomized, double-blind study. Nine mildly asthmatic subjects with a history of exercise induced asthma took part. This was conducted, first as a dose-response study and, second, as a time-course study. In the dose-response study, the provocative dose of saline-laden air causing a 25% fall in FEV1 was calculated (PD25). Terfenadine (180 mg) and the combination of terfenadine (180 mg) plus flurbiprofen (100 mg) both protected significantly against hypertonic saline challenge, achieving increases in PD25 values by factors of 7.24 and 6.30, respectively. Flurbiprofen (100 mg) also displaced the dose-response to the right, increasing the PD25 by a factor of 1.92, but this protection was significantly less than that afforded by terfenadine. In the time-course studies, a single inhalation of hypertonic saline previously shown to cause at least a 25% fall in FEV1 was administered, and FEV1, was followed for 30 min. Preadministration of terfenadine reduced the mean area under the curve of percentage fall in FEV1-time response by 68.5%, with similar results obtained with the combination of terfenadine and flurbiprofen. We conclude that the bronchoconstriction in asthma provoked by inhaled hypertonic saline is mediated predominantly through the hyperosmolar release of histamine from airway mast cells, with a minor contribution being made by prostanoids. PMID- 2571321 TI - Reversal of leukotriene D4- and leukotriene E4-induced airway constriction in the guinea pig. AB - Conscious Hartley guinea pigs were challenged with LTD4 or LTE4 aerosols. When dynamic compliance (Cdyn) decreased to 50% of its baseline value, the challenge aerosols were stopped and treatment aerosols of salbutamol, the LTD4/E4 antagonist LY171883 Na, atropine, or sodium chloride (control) were begun. After 5 min of continuous exposure to the treatment aerosol, each animal was killed and excised lung gas volume (ELGV) was measured. Salbutamol was equally effective in reversing LTD4- and LTE4-induced Cdyn changes. Aerosolized LY171883 Na was effective against both agonists, but was about threefold more potent against LTE4. In contrast, atropine was very effective in reversing LTD4-induced Cdyn changes, but produced minimal reversal when LTE4 was the challenge agent. Pulmonary gas trapping results supported these observations; ELGV values were closely correlated with 5-min Cdyn measurements for both LTD4 (r = -0.817) and LTE4 (r = -0.831). Thus, although LTD4 and LTE4 are though to act on the same or similar receptors, the pattern of pharmacologic reversal at comparable levels of airway obstruction differs for these two agonists. PMID- 2571322 TI - The extended digitorum brevis muscle flap. AB - The anatomy of the anterior tibial artery was studied in eight fresh cadavers. Consistently no major branches were found to the long toe extensor or anterior tibial muscle in the segment 15 cm preceding the lateral malleolus. This allowed the use of the extensor digitorum brevis muscle on a long anterior tibial vascular pedicle following release of the extensor retinaculum. The longer pedicle made this reliable muscle flap suitable for coverage of mid and lower tibial defects, in addition to ankle defects, as an alternative to free flaps. Its use is described in three complex cases. PMID- 2571323 TI - [Basic and clinical evaluation of an immunoradiometric competitive inhibition assay for sialyl TN antigen: (2). Evaluation of clinical significance. STN Study Group]. AB - The clinical significance of serum sialyl Tn antigen as a tumor marker was evaluated using "STN Otsuka" kits. Results indicated that the antigen was frequently elevated in sera from patients with ovarian cancers (43.1%, 140/325), including serous cystadenocarcinoma (51.6%, 63/122), mucinous cystadenocarcinoma (55.6%, 30/54), endometrioid carcinoma (56.5%, 13/23), and mesonephroid carcinoma (40.0%, 6/15). The positive frequency of sialyl Tn antigen in patients with ovarian carcinoma was less than the frequency of CA 125 (75.5%, 194/257) or sialyl SSEA-1 (47.2% 83/176). However, the false positive rate of sialyl Tn antigen in patients with benign gynecological disorders (3.7%, 15/401) was much lower than the false positive rates of other antigens; such as CA 125 (48.4%, 155/320) and sialyl SSEA-1 (19.2%, 51/266). The serum level of sialyl Tn antigen reflected the clinical activity of the disease quite well in patients with ovarian cancers. The sialyl Tn antigen was concluded to be a useful serum tumor marker for ovarian cancers. PMID- 2571324 TI - [Antiemetic efficacy of intravenous timiperone in emesis induced by a 5-day course of cisplatin]. PMID- 2571325 TI - Ultrastructure of Sertoli cells in cryptorchid goats. AB - Sertoli cells of intra-abdominal testes from 25 naturally occurring, unilaterally cryptorchid, West African pygmy goats between the ages of 1-30 months were morphologically examined by light and transmission electron microscopy. Ultrastructurally, Sertoli cells in the intra-abdominal testes from 1-month-old goats were columnar and contained ovoid nuclei. The cytoplasm depicted clusters of rough endoplasmic reticulum cisternae (RER). In 3- to 4-month-old animals nucleolonemas of the Sertoli cells were associated with vesicles; the cytoplasm contained RER and areas of whorls of smooth endoplasmic reticulum. An abundance of lipid droplets was accompanied by lipofuscin granules in the cytoplasm. In 6- to 8-month-old goats RER cisternae presented circular or irregular profiles in the cells with an augmentation of cytoplasmic lipid droplets and lipofuscin granules. In 12- to 15-month-old animals intercellular spaces in the seminiferous epithelium exhibited focal expansions. Nuclear profiles of the Sertoli cells were irregular. In 24- to 30-month-old animals the Sertoli cells were in an advanced stage of degeneration. PMID- 2571326 TI - Changing therapy from timolol to betaxolol. Effect on intraocular pressure in selected patients with glaucoma. Timolol-Betaxolol Study Group. AB - Three hundred fifty-three patients whose intraocular pressure was controlled with a timolol maleate ophthalmic solution were studied. Following a baseline period, half were switched (masked and randomized) to treatment with a betaxolol hydrochloride ophthalmic solution and were followed up for 12 weeks. Intraocular pressure, signs, and symptoms were recorded at weeks 1, 4, 8, and 12. Those patients switched to the betaxolol ophthalmic solution had a significant increase in both ocular side effects (burning/stinging and tearing) and intraocular pressure at weeks 4, 8, and 12 when compared with those patients who continued to receive timolol. PMID- 2571327 TI - Comparison of a noncardioselective beta-adrenoceptor blocker and a cardioselective blocker in reducing aqueous flow in humans. AB - In a double-masked crossover study, the dose-response relationship for aqueous flow was determined for four concentrations of betaxolol hydrochloride and levobunolol hydrochloride in 19 normal subjects. One eye of each subject received every concentration of both drugs. Each concentration was administered once daily for 1 week. The fellow eye received a placebo. Aqueous flow was measured for several hours on the last day of administration of each concentration beginning immediately after drug administration. For levobunolol, the mean decrease in aqueous flow compared with baseline was 8% after 1 week's treatment with 0.017% levobunolol, 15% after treatment with 0.05%, 20% after treatment with 0.167%, and 32% after treatment with 0.5%. For betaxolol, the mean decrease in aqueous flow compared with baseline was 3% after 1 week's treatment with 0.017% betaxolol, 12% after treatment with 0.05%, 18% after treatment with 0.167%, and 17% after treatment with 0.5%. Levobunolol-treated eyes but not betaxolol-treated eyes showed a significant drug effect 1 week after discontinuing the drug therapy. The relative potency of the noncardioselective drug was greater than that of the beta 1-cardioselective drug, but the difference in potency was much less than would be expected solely based on their relative affinity for beta 2-receptors in other species and tissues. PMID- 2571328 TI - Long-term follow-up of a kindred with multiple endocrine neoplasia type IIA. AB - A kindred with multiple endocrine neoplasia type IIA was studied. In four of five fully documented cases, patients presented with adrenal medullary disease during the second or third decade; in three cases, medullary thyroid carcinoma was not diagnosed until the fourth decade. This kindred seems to have an aggressive variant of adrenal medullary disease in which clinical presentation is early, metastasis occurs, and oncological cure is uncommon. Early detection and operation were important, and symptoms were controlled with operations and alpha- and beta-adrenergic blockade in uncured members. PMID- 2571329 TI - Lawn mower injuries in children. AB - Lawn mowers cause severe injuries, particularly to the lower limbs in children. The study of 52 inpatient cases treated over 12 years shows that ride-on lawn mowers cause the most severe trauma, resulting in longer hospitalization. These children more often require further admissions for reconstructive surgery including free tissue transfer. These accidents can be avoided if young children are prevented from playing near or using power lawn mowers. PMID- 2571330 TI - New approaches to the control of obesity in animals and their clinical potential. PMID- 2571331 TI - A novel ubiquitous protein 'chaperonin' supports the endosymbiotic origin of mitochondrion and plant chloroplast. AB - The deduced amino acid sequences for a major mitochondrial protein (designated P1, related to the 'chaperonin' family of proteins) from human and Chinese hamster cells show extensive similarity (greater than 60% identity observed over the entire length) with a related protein present in evolutionarily as divergent organisms as Escherichia coli, Coxiella burnetii, Mycobacterium species, cyanobacteria as well as in yeast mitochondria and higher plant chloroplasts. Of the different groups of bacteria for which sequence data is available, maximum similarity of the mammalian/yeast P1 protein is observed with the corresponding protein from purple bacteria (especially C. burnetii) while the protein from plant chloroplasts exhibited highest similarity with the corresponding protein from cyanobacteria. The sequence data for this protein thus support the contention that the endosymbiont that gave rise to mitochondrion was a member of purple bacteria, while plant chloroplast originated from a member of the cyanobacterial lineage. PMID- 2571332 TI - Cross-linking of lipocortin I and enhancement of its Ca2+ sensitivity by tissue transglutaminase. AB - The stimulation of human epidermoid carcinoma A431 cells with the calcium ionophore A23187 resulted in the formation of high-molecular-weight lipocortins I, having apparent molecular weights of 75 kDa and 160 kDa as detected with specific anti-lipocortin I antibody. These immunoreactive proteins were identified to be covalently cross-linked multimers of lipocortin I, since essentially the same cross-linked multimers were observed when purified lipocortin I was incubated with tissue transglutaminase (TGase) in vitro. Classical amine substrates for TGase, such as dansylcadaverine and putrescine, were also incorporated stoichiometrically into lipocortin I. Cross-linking or amine incorporation was not observed with lipocortin II. Des 1-26 lipocortin I did not serve as a substrate for TGase, indicating that the N-terminal region of lipocortin I plays an important role in the formation of lipocortin I multimers. The cross-linking of lipocortin I by TGase resulted in a remarkable enhancement of calcium sensitivity for phospholipid binding; i.e., the free calcium concentration required for the cross-linked lipocortin I to attain 50% maximal binding to phosphatidylserine vesicles was as little as 3 microM, while that required for intact monomeric lipocortin I was 20 microM. PMID- 2571333 TI - Daunorubicin and vincristine binding to plasma membrane vesicles from daunorubicin-resistant and wild type Ehrlich ascites tumor cells. AB - Tumor cell resistance to anthracyclines, epipodophyllotoxins and vinca alkaloids, called multi-drug resistance (MDR) is intimately linked to changes in the plasma membrane which facilitate an increased energy dependent drug extrusion in the resistant cell compared to the wild type cell. Isolated plasma membrane vesicles from wild type Ehrlich ascites tumor cells (EHR2) and the daunorubicin (DNR) resistant subline EHR2/DNR+ were utilised to study binding and possible transport of DNR and vincristine (VCR). A significant ATP enhanced increase in VCR binding to vesicles from EHR2/DNR+ compared to EHR2 was demonstrated. Furthermore, an increase in ATP enhanced VCR binding in proportion to content of the MDR associated P-glycoprotein was seen in plasma membrane vesicles prepared from various benign human endocrine tumors. VCR binding to EHR2/DNR+ vesicles was inhibited by other vinca alkaloids greater than actinomycin D greater than colchicine greater than anthracyclines, with 35-75 microM concentrations of anthracyclines needed for 50% inhibition. VCR binding to EHR2/DNR+ vesicles was pH and temperature dependent with an activation energy of -30 kJ/mol and was decreased by replacement of Na+ with K+ and by addition of Ca2+. Preincubation of vesicles with monoclonal antibody against the C terminal of P-glycoprotein had no effect on VCR binding and osmolality tests failed to show genuine transmembranal transport of VCR. DNR binding was similar in plasma membrane vesicles from both cell lines, and showed none of the characteristics mentioned for VCR. Furthermore, a radiolabeled N-hydroxysuccinimide ester derivative of doxorubicin, which inhibited VCR binding to EHR2/DNR+ membranes to an even greater extent than doxorubicin, labeled plasma membrane proteins from EHR2 and EHR2/DNR+ identically and did not demonstrate any binding to P-glycoprotein. Therefore, even though the study confirms the close link between vinca alkaloid binding and P-glycoprotein, it could not detect a similar association between anthracyclines and P glycoprotein thus attesting to the complexity of the MDR phenotype. PMID- 2571334 TI - Action of beta-adrenoceptor blockers on liver function of rats. AB - Three nonselective beta-adrenoceptor blockers, propranolol, cloranolol and talinolol, were examined in male rats. The amount and function of polysubstrate monooxygenase, serum bilirubin and carbohydrate metabolism were investigated. Doses producing a 25% reduction in heart beat/min were given orally. The effect of a single dose was compared to that of a 12-day treatment period. Propranolol in a single dose did not influence hepatic parameters. The 12-day treatment reduced the amount of cytochrome P-450 (cP-450) and prolonged hexobarbital biotransformation time. Cloranolol decreased cP-450, prolonged hexobarbital anaesthesia and inhibited aminopyrine-N-demethylation (AND) even in a single dose. No further impairment occurred with continued treatment. A single dose of talinolol led to cP-450 loss and inhibited cP-450 dependent liver functions. This inhibition progressed with continued treatment. High bilirubin levels were measured. Both cloranolol and talinolol elicited an initial rapid hyperglycaemia with normal liver glycogen content. At the end of the treatment blood glucose and liver glycogen values were normal. The reversible hyperglycemic reaction shows the necessity of controlling glucose tolerance. When talinolol is administered liver function parameters should be checked in appropriate time intervals. PMID- 2571335 TI - Inhibition of coronary artery transplant atherosclerosis in rabbits with angiopeptin, an octapeptide. AB - Accelerated coronary atherosclerosis of cardiac allograft occurs in 30-40% of cardiac transplant patients and remains an unsolved clinical problem. The etiology is unknown and anti-platelet drugs are used without conspicuous success. The inhibitory effect of the octapeptide, angiopeptin on coronary atherosclerosis was studied in a previously described rabbit heterotopic cardiac transplant model where allograft rejection is prevented by daily administration of cyclosporin A (CsA, 10 mg/kg per day s.c.). Twenty male New Zealand white rabbits (2.6-2.8 kg) received a heterotopic cardiac transplant from rabbits of the same strain. Donors and recipients were fed a 0.5% cholesterol diet 1 week prior to transplantation which was continued for the recipient until death 6 weeks later. The control group (n = 16) received CsA and saline injections twice daily and the treatment group (n = 4) received CsA and angiopeptin (60 micrograms/rabbit daily s.c.) in 2 divided doses. The treatment began after completion of the transplantation. Coronary artery transplant atherosclerosis was uniformly distributed (tubular) in the entire length of the coronary arteries. Angiopeptin inhibited the intimal hyperplasia in the transplanted heart from 47.5 +/- 2.4% (mean +/- SE) to 25.0 +/ 6.9% and in the native heart from 24.2 +/- 1.4% to 15.7 +/- 1.5%. The intimal hyperplasia is expressed as area of intimal hyperplasia/total vessel area x 100%. A similar inhibition by angiopeptin was seen in lipid deposition in the donor ascending aorta which is transplanted with the heart. Angiopeptin attenuated significantly the hyperplasia and the lipid deposition of the native coronary arteries and aorta but to a lesser extent.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571336 TI - Studies of anti-F(ab')2 antibodies and VH region distribution among SLE kindreds. AB - Genomic DNA samples from kindreds of patients with systemic lupus erythematosus were examined for VH region genotypes using probes representing JH and representative members of the human VH gene families (VHI-VHVI) in parallel with distribution of high levels of anti-F(ab')2 (generic anti-idiotype) in some non affected SLE family members. Identical VH region genotypes were present in an SLE female proband and one unaffected normal male sibling. Three normal unaffected family members with elevated anti-F(ab')2 showed AB, AD, and AC (non-identical)VH genotypes. VH region genotype pattern distributions do not appear to directly correlate with production of high levels of anti-F(ab')2 in SLE families. PMID- 2571337 TI - Genetic studies in Sjogren's syndrome and systemic lupus erythematosus. AB - HLA associations with Sjogren's syndrome (SS), previously defined by serologic HLA typing, are reviewed. Because the SS-A/Ro and SS-B/La autoantibody responses in SS and systemic lupus erythematosus (SLE) show even stronger correlations with HLA alleles, HLA-DR and DQ alleles were examined using restriction fragment length polymorphisms (RFLP) in white and black patients with SS and/or SS having anti-Ro and anti-La antibodies. The strongest associations across ethnic lines were with HLA-DR3, DQw2 and DQw1.2 (DQw6), especially the DQw1.2 (DQw6)/Dqw2 heterozygous state. HLA-DQw3 was relatively decreased. These data suggest that HLA-DQ is most likely responsible for these abnormal autoantibody responses. In addition, multiplex family studies of Ro antibodies demonstrates the requirement for both HLA and non-MHC effects. PMID- 2571338 TI - Viral genomes in lymphomas of patients with Sjogren's syndrome. AB - The recent isolation of a new member of the herpes virus family (Human Herpes Virus-6, HHV-6) from patients with lymphoproliferative diseases prompted us to examine biopsies from six patients with primary Sjogren's Syndrome (SS) who developed non-Hodgkin's lymphoma. Five SS patients developed B-cell lymphoma and one developed a T-cell lymphoma based on immunoglobulin and T-cell antigen receptor (TCAR) gene rearrangements. In two SS patients with B-cell lymphomas, viral DNAs were detected, including: (a) Epstein-Barr Virus (EBV) DNA that exhibited an unusual pattern of restriction fragment length polymorphisms (RFLP) of the Bam M viral DNA segment; and (b) HHV-6 DNA in a second SS patient's lymphoma, with an RFLP similar to recent viral isolates from patients with other lymphoproliferative diseases. Viral DNA was not detected in the other four SS lymphoma biopsies. Also, all six biopsies were examined for presence of other viral DNAs (including CMV, HTLV, HIV and adenovirus) and were negative. Antibody titers to EBV-associated early-diffuse-antigen (EA-D), as assessed by ELISA method, and antibody titers against HHV-6, as detected by immunofluorescence and radio-immunoprecipitation assays, were markedly elevated in several SS patients with lymphoma and pseudolymphoma. These results suggest a potential role of EBV or HHV-6 in the neoplastic transformation that occurs with increased frequency in SS patients. PMID- 2571339 TI - Are antagonists of dopamine D1 receptors drugs that attenuate both positive and negative symptoms of schizophrenia? A pilot study in Java monkeys. AB - Amphetamine is known to elicit stereotyped behavior in various species. For a long time, this effect was considered to be an animal model for the positive symptoms of schizophrenia. In addition, amphetamine is known to induce a strong social isolation in socially living monkeys. Both on symptomatologic and pharmacologic grounds, this amphetamine-induced social isolation has been suggested to represent an animal model for the negative symptoms of schizophrenia. To date no effective treatment has been found for these negative symptoms. We now report that the selective D1 dopamine antagonist SCH 23390 is very effective in antagonizing both the stereotyped behavior and the social isolation in Java monkeys induced by amphetamine. Moreover, SCH 23390 is able to reinstate normal behavior in these animals. These results may have important consequences for our understanding of the functional significance of the D1 receptor as well as for the clinical treatment of the positive and negative symptoms of schizophrenia. PMID- 2571340 TI - Electrophysiological effects of selective sigma-receptor agonists, antagonists, and the selective phencyclidine receptor agonist MK-801 on midbrain dopamine neurons. AB - Extracellular single unit recording techniques were used to study the effects of selective sigma-receptor agonist [(+)-3-PPP, (+)-pentazocine, and DTG] and selective sigma-receptor antagonists (BMY 14802 and Rimcazole) on dopamine neurons of the substantia nigra. Intravenous (IV) administration of sigma agonists decreased, whereas IV administration of the sigma antagonist BMY-14802 increased the firing rate of dopamine neurons. The other sigma antagonist Rimcazole produced inconsistent changes in dopamine unit activity. These data, in conjunction with anatomic data suggesting sigma receptor localization on dopamine neurons in the substantia nigra (Gundlach et al: J Neurosci 6:1757-1770, 1986; Graybiel et al: Soc Neurosci Abstr 13:28, 1987) demonstrate a relationship of the sigma receptor with the dopamine system and further suggest a model system to study agonist-antagonist interactions of sigma ligands. The selective phencyclidine (PCP) agonist MK-801 was equipotent to PCP in regard to stimulatory properties on dopamine neurons. However, the relative potencies do not correspond to their relative binding affinities, suggesting that non-PCP-receptor properties may mediate this effect. PMID- 2571341 TI - New antagonists of glutamate receptors. PMID- 2571343 TI - Transglutaminase in membranes rich in nicotinic acetylcholine receptors. PMID- 2571344 TI - Purification and properties of brain neuroleptic-binding proteins. PMID- 2571342 TI - Isolation of argiopinin-binding protein from bovine brain. PMID- 2571346 TI - Identification of partial mutational sites in human mitochondrial DNA in the Chinese and its significance. AB - A simple method for the identification of mutational sites in human mitochondrial DNA (mtDNA) was described. It was based on the human Cambridge sequence as a relative standard sequence and a single base pair substitution in mtDNA as a unique mutational form. The partial mutational sites can be determined using this method which was characterized by combining the restriction mapping with the analysis for the table of human mtDNA potential mutational sites with rapidity and simplicity. In the meanwhile, six mtDNA mutational sites found in Chinese population were identified by means of this method. PMID- 2571345 TI - Regulation of synapsin I phosphorylation by cellular modulators. PMID- 2571347 TI - Postembedding immunogold labelling of fixed glutamate: an electron microscopic analysis of the relationship between gold particle density and antigen concentration. AB - A series of glutamate-glutaraldehyde-brain protein conjugates, prepared with different glutamate concentrations, was embedded in resin, cut, and incorporated in a multilayered sandwich which was embedded anew. Ultrathin cross-sections through the sandwich were collected on mesh grids and incubated together with tissue sections in an antiserum recognizing fixed glutamate, followed by a second layer antibody coupled to colloidal gold particles. This model system revealed a roughly linear relationship between the concentration of fixed glutamate (up to 154 mmol/l) and gold particle density. Extrapolated to the accompanying tissue sections this finding implies that a tissue compartment displaying 2n gold particles/microns2 contains twice as many fixed glutamate residues per volume as a compartment with n gold particles/microns2. By use of the model system the concentration of fixed glutamate in presumed glutamatergic nerve terminals in the cerebellum was estimated to be two to three times the average tissue level and about five times higher than the concentration of fixed glutamate in terminals thought to use gamma-aminobutyric acid as transmitter. This study represents a step towards the use of electron microscopic immunocytochemistry as a tool to assess the absolute concentrations of neuroactive amino acids in nerve terminals and other tissue compartments. PMID- 2571348 TI - Appearance of predictors of disease progression in relation to the development of AIDS. AB - To study the natural history of HIV-1 infection in relation to serological and immunological profiles, 199 asymptomatic HIV-1-antibody (HIV-1-core-antibody) seropositive and 76 seroconverted homosexual men were followed prospectively for 39 months. AIDS was diagnosed in 38 men. The AIDS attack rate was 20.8% after 39 months. The AIDS attack rate in the HIV-I-core-antibody positives was 12.1, versus 30.1% in the HIV-1-core-antibody negatives (P less than 0.001), and it was 13.3% in the HIV-1-antigen (HIV-1-Ag) negatives versus 53.9% in the HIV-1-Ag positives (P less than 0.001). The AIDS attack rate after 39 months was 10.9% in men with counts greater than or equal to 0.5 x 10(9)/l and 49.9% in those with CD4+ lymphocyte counts less than 0.5 x 10(9)/l. AIDS attack rates after 30 months in the same cohort have been previously reported [1], and were as follows: 6.8% in the core-antibody positives versus 35.7% in the core-antibody negatives. 6.9% in the HIV-1-Ag negatives versus 43.9% in the HIV-1-Ag positives, and 6.1% in those with CD4+ lymphocyte counts greater than or equal to 0.5 versus 51.9% in those with CD4+ lymphocyte counts less than 0.5 x 10(9)/l. The disappearance of core antibody, the appearance of antigen and the occurrence of low CD4+ lymphocyte counts preceded AIDS by a mean (s.d.) of 21.3 (8.9), 17.7 (8.8) and 15.7 (8.9) months, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571350 TI - Antibodies in sera from Norwegian multiple sclerosis patients and the general population reacting with HTLV-1. A pilot study. AB - By using enzyme-linked immunosorbent assay, Western blot and passive agglutination we have performed a pilot study involving the testing of 349 sera from the general population of 3 coastal regions in southern Norway, 230 sera from multiple sclerosis patients from western Norway, 109 age- and sex-matched controls and 34 sera from patients with other neurological diseases, for antibodies to HTLV-I. About 2% and 4% of sera from the general population had antibodies reacting with HTLV-I in ELISA and Adult T-cell Leukemia Antigen by agglutination, respectively, whereas about 8% and 5% of the multiple sclerosis sera had such antibodies. In the general population antibodies were more frequent among young individuals and there appeared to be some prevalent regional differences. Western blot confirmatory tests showed that in some instances antibodies reacted with the gag proteins, in particular p19. The results obtained in this study do not indicate an association between HTLV-I and multiple sclerosis, although at this stage we cannot rule out that another retrovirus is involved. PMID- 2571349 TI - Progression to AIDS in the majority of asymptomatic HIV-infected people. AB - Sixty-eight asymptomatic HIV-seropositive people with a CD4 lymphocyte count above 400/mm3 at the first examination were followed up every year over a 3-year period, by monitoring the biological markers of AIDS (CD4 lymphocyte decrease, loss of anti-p24 or anti-p17 antibodies, positive p24 antigenemia, increase of erythrocyte sedimentation rate, and of serum levels of immunoglobulin G. immunoglobulin A, neopterin and beta 2-microglobulin). The percentages of subjects positive for at least one marker at the first, second, third and fourth examinations were 66, 88, 94 and 97%, respectively. The increase in the number of markers with time was significant (chi-square test; P less than 0.001). This increase suggests a progression to AIDS in the majority of asymptomatic seropositive subjects, even those without a decreased CD4 lymphocyte count. PMID- 2571351 TI - Increased adherence to vaginal epithelial cells and phagocytic killing of gonococci and urogenital meningococci associated with heat modifiable proteins. AB - Urogenital Neisseria meningitidis were characterized with regard to serogroup, colony morphology, the presence of heat modifiable proteins (HMP), attachment to human vaginal and buccal epithelial cells, and phagocytic killing by polymorphonuclear leukocytes. The findings were compared with those on gonococci, and with those on meningococci isolated from blood or cerebrospinal fluid, with regard to colony morphology, HMP and piliation. The opacity colony morphology characteristic could be used to predict the presence of HMP in gonococci but not in meningococci, and sodium dodecyl sulphate polyacrylamide gel electrophoresis had to be used to demonstrate this surface protein. The urogenital meningococci, serogroup Y, attached significantly more efficinetly to vaginal epithelial cells in the presence of HMP and behaved in this respect like those of gonococci. Gonococci and meningococci containing HMP were more sensitive to phagocytic killing than those without HMP. Meningococci from opaque and transparent colonies and isolated from patients with meningococcal disease had no demonstrable HMP. They showed low adherence to vaginal and buccal epithelial cells, with no difference between organisms from opaque or transparent colonies. PMID- 2571352 TI - Association of myocardial glutamate and aspartate pool and functional recovery of postischemic heart. AB - The effect of low flow ischemia and subsequent reperfusion with 5.5 mM glucose or 5 mM acetate on energy metabolism and catabolism of myocardial glutamate and aspartate was studied in isolated perfused guinea pig hearts. Reperfusion with acetate was followed by low recovery of the cardiac contractile function associated with a great rise in isovolumic end-diastolic pressure. It was combined with more profound losses of tissue adenine nucleotides and the total Cr compared to reperfusion with glucose. The total glutamate and aspartate pool decreased more than two-fold compared to the initial one regardless of substrate. However, glutamate content was reduced by 58 and 38% with acetate and glucose, respectively. The expenditure of both amino acids was caused by alanine formation stimulated by glycolysis/glycogenolysis. The remaining glutamate and aspartate pool in the reperfused hearts positively correlated with adenine nucleotides (r = 0.62), the total creatine (r = 0.65), and the recovery of contractile function (r = 0.64). The results suggest that the glutamate and aspartate pool may be of critical importance for postischemic functional and metabolic recovery of the heart. PMID- 2571353 TI - End products of glutamine oxidation in MC-29 virus-induced chicken hepatoma mitochondria. AB - Products of glutamine metabolism were examined in the MC-29 virus-induced chicken hepatoma mitochondria incubated in vitro. Glutamine oxidation proceeded in the tumor mitochondria exclusively via a pathway involving glutamic-oxalacetic transaminase. Malate stimulated aspartate production from glutamine, while pyruvate exerted suppressive effect on aspartate production with little alanine formation. The mitochondria of this hepatoma are unique in that the metabolic pattern and response to malate and pyruvate are essentially inconsistent with those reported in normal cells as well as those proposed by Moreadith and Lehninger in various tumor cells. PMID- 2571354 TI - The benefits of prescription information leaflets (2). AB - Prescription information leaflets (PILs) for penicillins, diuretics and benzodiazepines were evaluated in a small Hampshire town. A second town, in which no leaflets were distributed, acted as a control. Knowledge about these medicines was greater amongst the 252 patients who received leaflets compared with 247 controls. Significantly more patients who received leaflets knew the potential side-effects of their treatment and, with the exception of those taking penicillins, were more satisfied with the information they received. These findings add further support for the routine use of PILs in general practice. PMID- 2571355 TI - Amplification and expression of mdr1 gene in a multidrug resistant variant of small cell lung cancer cell line NCI-H69. AB - Amplification and expression of the mdr1 gene encoding P-glycoprotein have been studied in H69/LX4 a multidrug resistant variant (MDR) of small cell lung cancer (SCLC) cell line NCI-H69. Recently a second independently derived MDR variant of this cell line designated H69/AR was found by others not to show amplification, rearrangement or over-expression of the mdr1 gene. The present study reports that in marked contrast to H69/AR, H69/LX4 shows amplification and expression of the P glycoprotein gene and raises the possibility that P-glycoprotein hyperexpression may be a clinically relevant component of MDR in some SCLC tumours. PMID- 2571356 TI - An evaluation of immunoreactivity for c-erbB-2 protein as a marker of poor short term prognosis in breast cancer. AB - Eighty-five breast carcinomas from the same number of patients have been assessed immunohistochemically using the antiserum 21N for the presence of the c-erbB-2 protein. Twenty-two of the patients had evidence of advanced disease (tumour fixation or distant metastases) at presentation. Follow-up was for a median of 24 months. c-erbB-2 protein was detected in the majority of cells in 14 (16.5%) carcinomas, and to a lesser extent in a further six (7%) tumours. There was no relationship between staining and stage, node status or size but more poorly differentiated carcinomas had evidence of staining (36%) than well (17%) or moderately (14%) differentiated carcinomas (P = 0.02). There was a significant association between staining and mortality (P = 0.009) and recurrence (P = 0.0002). The relative risk of death for staining compared to no staining (after adjusting for node status, stage and grade) was 2.97 (95% confidence interval 1.29, 6.84) and the relative risk of recurrence for staining compared to no staining after similar adjustment was 3.85 (95% confidence interval 1.86-7.97). In this particular group of patients immunoreactivity for c-erbB-2 protein is an independent indicator of poor short-term prognosis. PMID- 2571357 TI - Partial unfolding of dodecameric glutamine synthetase from Escherichia coli: temperature-induced, reversible transitions of two domains. AB - Glutamine synthetase (GS), Mr 622,000, from Escherichia coli contains 12 active sites formed at heterologous interfaces between subunits [Almassy, R. J., Janson, C. A., Hamlin, R., Xuong, N.-H., & Eisenberg, D. (1986) Nature (London) 323, 304 309]. Temperature-induced changes in UV spectra from 3 to 68 degrees C were reversible with the Mn2+- or Mg2+-enzyme at pH 7.0 (50 degrees C) in 100 mM KCl. No dissociation or aggregation of dodecamer occurred at high temperatures. The thermal transition involves the exposure of approximately 0.7 of the 2 Trp residues/subunit (by UV difference spectroscopy) and 2 of the 17 Tyr residues/subunit (change in exposure from 4.7 to 6.7 Tyr/subunit by second derivative spectral analysis). Monitoring changes in Trp and Tyr exposure independently gives data that conform to a two-state model for partial unfolding with Tm values (where delta G unfolding = 0) differing by 2-3 degrees C at each level of [Mn2+] studied and with average delta HvH values of 80 and 94 kcal/mol, respectively. These observations suggest that two regions of the oligomeric structure unfold separately as independent transitions (random model). However, the data can be fit equally with a sequential model in which the Trp transition occurs first upon heating. By fitting with either model, Tm values increase from approximately 47 to approximately 54 degrees C with increasing free [Mn2+] from 3.6 to 49 microM but decrease from approximately 54 to approximately 43 degrees C by further increasing free [Mn2+] from 0.05 to 10 mM; such behavior indicates that the high-temperature form of the enzyme binds Mn2+ more weakly but has more binding sites than the native enzyme. The high-temperature Mn-enzyme form is somewhat less unfolded than is the catalytically inactive apoenzyme, which undergoes no further Trp or Tyr exposure on heating and therefore is assumed to be the high-temperature form of divalent cation-free GS. Adding substrates [ADP, L-Met-(SR)-sulfoximine, Gln, Gln + NH2OH, or Gln + ADP] to Mn.GS increased Tm to varying extents by preferential binding to the folded form. Indeed, the transition-state analogue complex GS.(Mn2.ADP.L-Met-(S)-sulfoximine phosphate)12 was stable in the folded form to at least 72 degrees C. Moreover, an Arrhenius plot for gamma-glutamyl transfer activity was linear from 4 to 72 degrees C with Ea = 18.3 kcal/mol.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2571358 TI - The assessment of relative surface hydrophobicity as a factor involved in the activation of human polymorphonuclear leukocytes by uropathogenic strains of Escherichia coli. AB - The initiation of the respiratory burst and the degranulation of human polymorphonuclear leukocytes (PMN) in response to stimulation by uropathogenic strains of Escherichia coli is dependent on the expression of Type 1 fimbriae by those strains. These PMN responses correlate with an increasing tendency of the interacting E. coli strain to be retained on hydrophobic columns. The present work assessed the measurement of relative surface hydrophobicity in relation to PMN activation. Type 1 fimbriate organisms bound most readily to Octyl-Sepharose columns and were strongly agglutinated in the salt aggregation test. In contrast, the same organisms partitioned to the dextran-rich (hydrophilic) phase of aqueous two-polymer phase systems. Electron microscopic observation of the organisms eluted from the Octyl-Sepharose columns and of the organisms recovered from both phases of the aqueous two-phase systems demonstrated, however, that both Type 1 and P-fimbriate organisms were retained on the columns and partitioned into the dextran-rich phase as a consequence of their being fimbriate and failed to identify this as a major factor in the activation of PMN. In addition electron microscopy demonstrated that each P-fimbriate population had fewer organisms expressing fimbriae than did Type 1 fimbriate populations, confirming the importance of phase variation as a factor affecting the physicochemical characteristics of a bacterial population. PMID- 2571360 TI - Constructive integration of learning theory and phenomenological approaches to biofeedback training. AB - Two major sources of theoretical development for biofeedback as an intervention paradigm are considered. An integration of cognitive learning theory approaches to the potential regulation of autonomic processes in an information-processing framework and the phenomenological information-processing approach of Kelly's personal construct theory suggest a new methodological paradigm for biofeedback as a tool of psychotherapeutic intervention, especially for the discipline of behavioral medicine. Biofeedback is reconstructed as a sequence of allocating attention to automatic cognitive processes until cognitive control has been mastered. This sequence is also seen as a circumspection-preemption-control cycle that Kelly (1955) suggested was essential to all problem solving. In light of Kelly's fundamental assumptions regarding the nature of constructs, it is suggested that controlled processing approaches to biofeedback require the biofeedback trainee to investigate both ends of psychophysiological dichotomies instead of demonstrative constructs of traditional biofeedback methodology. Other psychotherapeutic techniques are reviewed to validate this new theoretical approach. Finally, treatment within this paradigm is discussed as a recircumspection of relevant constructs that were routinized during the alarm reaction phase of Selye's general adaptation syndrome. PMID- 2571359 TI - Stress management and Gilles de la Tourette's syndrome. AB - Tourette's syndrome is a lifelong disorder characterized by multiple motor and verbal tics. The present study examined relaxation training and desensitization training as a method of reducing the frequency and intensity of tics and the distress they caused in a young adult diagnosed with Tourette's syndrome. After a period of symptom monitoring the subject underwent 3 weeks of intensive training in relaxation skills and 5 weeks of desensitization training with situational cues previously identified as eliciting Tourette's symptoms. According to self report monitoring, the experience of symptoms was decreased across 3 global dimensions: distress (48%), frequency (48%), and intensity (50%), and an hourly symptom count (50%). Collateral parental symptom report agreed with an observed decreased across distress (40%), frequency (41%), and intensity (40%). Inspection of data suggests that both components of stress management added to total treatment efficacy. PMID- 2571361 TI - [Regulation of Na,K-ATPase by synaptosomal factors and neurotransmitters]. AB - The identical increase of Na, K-ATPase activity is caused by oxidated and reduced forms of noradrenaline, serotonin and dopamine through the synaptosomal activating factors. The synaptosomal inhibiting factor, orthovanadate and calcium ions independently inhibit Na, K-ATPase activity. The inhibition constant (Ki) for vanadate does not change in the presence of EDTA, whereas in the presence of synaptosomal factors regulating the Na, K-ATPase factors, noradrenaline causes drastic increase of Ki for vanadate. It has been concluded, that the data point to the existence of special regulating system of brain synaptosomal Na, K-ATPase. PMID- 2571362 TI - Excitatory amino acids and neuropsychiatric disorders. PMID- 2571363 TI - Characteristics of the in vivo RBC: plasma lithium ratio in a clinical setting. PMID- 2571364 TI - ADH secretion in schizophrenic patients on antipsychotic drugs. AB - The authors studied antidiuretic hormone (ADH) secretion in schizophrenics on antipsychotic drugs. The mean plasma level of ADH is lower among schizophrenics than among controls at comparable values of plasma osmolality. In regression analysis of the relationship between plasma ADH and plasma osmolality, the slope of the regression line is gentler and the threshold of ADH secretion is lower in schizophrenics than that in control subjects. The authors suggest that the relationship between plasma ADH and plasma osmolality may be different in schizophrenics receiving antipsychotic drugs than in controls. PMID- 2571365 TI - [Catecholamine-containing sympathetic spinal neurons innervating the feline heart]. AB - The present study determined that a population "nonclassical" sympathetic neurons in cats spinal cord contains catecholamines. They are localized in the central, dorsomedial, and lateral regions of the ventral horn of T1-T5 segments of the spinal cord. Electrophysiological study indicated that axonal conduction velocity is 7.3 +/- 0.5 m/s (ranging from 3.6 to 17.2 m/s). Possible functional roles of catecholamine-containing neurons of spinal cord include involvement in sympathetic control of cardiac cycle duration. PMID- 2571366 TI - [Urinary enzymes induced in rats by intensive physical loading and the effect of the antioxidant potassium fenozan]. AB - The rats subjected to repeated heavy exercises (swimming with the load equal to 10% of body weight) had an increased activity of gamma-glutamyl transpeptidase in the urine. Both the percentage of the rats with hyperenzymuria and the level of enzyme were the highest after the fifth swimming. Water-soluble antioxidant (potassium fenozan) administered intraperitoneally 30 minutes before the swimming abolished the increase of enzymuria and maintained the enzyme activity to the normal level. PMID- 2571367 TI - [Pharmacological analysis of memory disorders of different origins]. AB - The influence of nootropic drugs of different chemical structure and mechanism of action (piracetam 200-800 mg/kg, meclofenoxate 50-100 mg/kg, nicergoline 1-4 mg/kg) on amnestic disturbances induced with electroconvulsive shock (ECS), scopolamine (2.5 mg/kg) and phenazepam (2.0 mg/kg) was studied in mice using passive avoidance test. All the investigated drugs possess distinct protective properties concerning different models of amnesia. In ECS-induced amnesia meclofenoxate was most effective, in scopolamine-induced amnesia--meclofenoxate and nicergoline, in phenazepam-induced amnesia all drugs were equally potent in profundity. It is supposed that neurophysiological mechanism of action of nootropic drugs is connected with directed activation of weakened memory trace of different etiology and its transmission into form receivable for transcription. PMID- 2571368 TI - [Isolation and molecular cytogenetic characteristics of the alpha-satellite DNA of human chromosome 6]. AB - In this work a molecular-cytogenetic characteristic has been given to the alpha satellite DNA of chromosome 6. Using different restrictases made it possible to evaluate structural heterogeneity of the obtained subgroup of the alphoid DNA. Polymorphic restriction fragments of DNA have been found which can be used in determining linkage groups. PMID- 2571369 TI - [The dynamic morphofunctional state of the adrenals in irradiated recipients following bone marrow transplantation]. AB - The influence of syngenic bone marrow transplantation in the dose of 1.10(7) cell/ml upon the adrenal gland state of lethally irradiated recipients at various stages of posttransplantation period during 3 months has been studied at 150 linear F1 (CBA x C57B) male mice. Histological and histochemical studies conducted have shown the transplantation of native and cryopreserved bone marrow of the lethally irradiated recipients to result, depending upon observation time, in undulating change in adrenal gland weight, cortex thickness and lipid content in spongiocytes, which testifies to the certain dynamics of processes, occurring in recipients' adrenal gland. PMID- 2571370 TI - Use of scid mice to identify and quantitate lymphoid-restricted stem cells in long-term bone marrow cultures. AB - Mice homozygous for an autosomal recessive scid (severe combined immune deficiency) mutation on chromosome 16 exhibit a defect that specifically impairs lymphoid differentiation but not myelopoiesis. Consequently such mice are deficient in both humoral and cell-mediated immune functions. Despite their defect, scid mice survive under pathogen-free conditions and are fertile. The mutation does not impair the hematopoietic microenvironment necessary for lymphoid differentiation, since these mice can be cured with grafts of normal bone marrow (BM) or cells from long-term BM cultures (LTBMC); however, reconstitution requires sublethal (400 cGy) irradiation of recipients. Engraftment with cells from LTBMC gave near-normal levels of colony-forming B cells (CFU-B) in spleen and BM of the recipients by 6 weeks postgrafting. Since LTBMC are devoid of all mature B and pre-B cells but contain stem cells that restore lymphoid function in scid mice, we used a limiting-dilution assay to characterize and enumerate the number of stem cells in LTBMC capable of restoring lymphoid function. Curing was determined by the CFU-B-cell assay, since CFU-B are not detectable in normal scid mice. The results indicate that fewer cells from LTBMC than from fresh BM are required to obtain lymphoid reconstitution. As few as 10(3) LTBMC cells can repopulate significant B- and T-cell function in scid recipients. From these results we conclude that scid mice can be used as recipients to quantify lymphoid-restricted stem cells and that there is a functional separation of lymphoid- and myeloid-restricted stem cells in LTBMC with an enrichment for lymphoid-restricted stem cells in these cultures. PMID- 2571371 TI - Involuntary administration of medication in the community: the judicial opportunity. AB - In a state in which patient refusal of antipsychotic medication in all nonemergency situations must be respected, lawyers and psychiatrists in western Massachusetts have employed probate court decisions as involuntary outpatient treatment orders. The legal, administrative, and clinical issues in sustaining court-ordered outpatient treatment are discussed by focusing on case examples demonstrating some successes, some challenges, and some failures. Judicially sanctioned involuntary outpatient treatment presents an alternative model to statutorily based outpatient commitment. PMID- 2571372 TI - Status of the gamma-glutamyl transpeptidase enzyme activity in mouse skin exposed to polyaromatic hydrocarbons and tumor promotor TPA. PMID- 2571373 TI - Effects of PCBs on plasma enzymes, testosterone level, and hepatic xenobiotic metabolism in the grey partridge, Perdix perdix. PMID- 2571374 TI - Carbohydrate deficient transferrin: a marker for alcohol abuse. AB - OBJECTIVE: To assess the value of serum carbohydrate deficient transferrin as detected by isoelectric focusing on agarose as an indicator of alcohol abuse. DESIGN: Coded analysis of serum samples taken from patients with carefully defined alcohol intake both with and without liver disease. Comparison of carbohydrate deficient transferrin with standard laboratory tests for alcohol abuse. SETTING: A teaching hospital unit with an interest in general medicine and liver disease. PATIENTS: 22 "Self confessed" alcoholics admitting to a daily alcohol intake of at least 80 g for a minimum of three weeks; 15 of the 22 self confessed alcoholics admitted to hospital for alcohol withdrawal; 68 patients with alcoholic liver disease confirmed by biopsy attending outpatient clinics and claiming to be drinking less than 50 g alcohol daily; 47 patients with non alcoholic liver disorders confirmed by biopsy; and 38 patients with disorders other than of the liver and no evidence of excessive alcohol consumption. INTERVENTION: Serial studies performed on the 15 patients undergoing alcohol withdrawal in hospital. MAIN OUTCOME measure--Determination of relative value of techniques for detecting alcohol abuse. RESULTS: Carbohydrate deficient transferrin was detected in 19 of the 22 (86%) self confessed alcohol abusers, none of the 47 patients with non-alcoholic liver disease, and one of the 38 (3%) controls. Withdrawal of alcohol led to the disappearance of carbohydrate deficient transferrin at a variable rate, though in some subjects it remained detectable for up to 15 days. Carbohydrate deficient transferrin was considerably superior to the currently available conventional markers for alcohol abuse. CONCLUSION: As the technique is fairly simple, sensitive, and inexpensive we suggest that it may be valuable in detecting alcohol abuse. PMID- 2571375 TI - Electrophysiology of the swimming system of hydromedusae and the effects of atropine-induced crumpling. AB - 1. In order to evaluate how crumpling behavior influences the swimming system in Liriope tetraphylla, extracellular recordings of muscle action potentials related to both behaviors were performed. 2. The swimming pattern of Liriope consists of irregular bursts of muscle potentials. A correlation was obtained between number of muscle potentials and burst length and between burst and interburst periods. The swimming pattern could be predicted from the burst length since a long burst was followed by a short period of quiescence and by a burst as long as the preceding one. 3. The addition of atropine to induce radial muscle contraction first caused an increase in swimming activity and then an irreversible reduction. The enhancement of swimming activity was due to the increase of burst length and of the frequency of swimming bursts (due to the decrease of the interburst period), and the reduction of swimming activity, to the enhancement of the interburst period and to the reduction of the burst length. 4. It is proposed that, when facing a noxious stimulus, Liriope will first "escape" from it by enhancing its swimming activity and, if the stimulus persists, blockade of the pacemaker system becomes accentuated as the radial muscle contractions become more sustained. PMID- 2571376 TI - Restriction fragment length polymorphism analysis with a cross-reactive HLA class II DR-beta gene probe for the detection of engraftment of MHC-mismatched marrow in the rhesus monkey. AB - Our interest in MHC-mismatched allogeneic bone marrow transplantation (BMT) in the rhesus monkey prompted us to investigate restriction fragment length polymorphism analysis as a means for detecting lymphohematopoietic chimerism in the primate. A human MHC (HLA) class II DR beta gene cDNA probe was tested on rhesus peripheral blood mononuclear cell DNA digested with any of three restriction enzymes. We found that (1) the human DR beta probe hybridized to as many as 15 restriction fragments per rhesus DNA sample, suggesting cross hybridization at more than one locus of rhesus MHC class II beta genes; (2) restriction fragment length polymorphisms were common among outbred monkeys as a result of class II beta gene polymorphisms and would be sufficient for chimerism detection in the majority of random pairs of outbred monkeys utilizing only a single restriction enzyme (Bgl II); and (3) sensitivity (5-10% chimerism) was comparable to that reported for restriction fragment length polymorphism assays utilizing non-MHC probes in clinical HLA-identical BMT. Utility of the assay was demonstrated in a preliminary series of experiments in rhesus monkeys conditioned with mixed T cell-depleted MHC-mismatched allogeneic plus T cell-depleted autologous BMT with or without cardiac allograft implantation. PMID- 2571377 TI - A comparison of DNA-RFLP typing with serology and mixed lymphocyte reaction in the selection of matched unrelated bone marrow donors. AB - Eleven leukaemic patients and 43 potential unrelated marrow donors were typed serologically, tested in mixed lymphocyte reaction (MLR) and typed by restriction fragment length polymorphism (RFLP) analysis. RFLP typing data were compared with DR serology and MLR results. Eleven of the 54 individuals showed discrepancies between DR serology and RFLP DR assignment. RFLP DR/DQ mismatch always correlated with positive MLR, but RFLP identity was present in both MLR negative and MLR positive pairs. RFLP typing cannot reliably predict a negative MLR response, but we suggest that it should be used in the selection of unrelated marrow donors to exclude mismatched donors from testing in the MLR. This will facilitate donor searches by reducing the number of MLR performed. PMID- 2571378 TI - Autotransplantation using peripheral blood stem cells mobilized by cyclophosphamide. PMID- 2571379 TI - Sorbitol-gelatin and glutamic acid-lactose solutions for stabilization of reference preparations of measles virus. AB - The work reported here sought to assess the protection afforded by two stabilizing solutions (sorbitol-gelatin and glutamic acid-lactose) in preserving the potency of freeze-dried Schwarz strain measles virus during storage with a view to the production of reference preparations and working lots of virus suspensions. Stabilized virus suspensions and control suspensions were stored at 70 degrees C or were freeze-dried and stored at -20 degrees C, and their potency was determined over a storage period of 21 months. It was found that the sorbitol gelatin imparted more satisfactory stability (r = +0.18) to the freeze-dried virus suspensions than did the glutamic acid-lactose. The results also indicate that sorbitol-gelatin, used under the conditions of this study, is an effective stabilizer in the preparation of freeze-dried suspensions of Schwarz strain measles virus employed as reference preparation working lots. PMID- 2571380 TI - Assessment and physical treatment of affective disorder. AB - Over the past decade there has been a tendency to diagnose affective disorder more readily, aided by the development of operational schedules that require few symptoms for diagnosis. This review will cover recent developments in the classification of affective disorder, how depression and mania can be adequately assessed and the treatment of these conditions. PMID- 2571381 TI - Inhibition of neurally-evoked transmitter release by calcium channel antagonists in rat parasympathetic ganglia. AB - 1. Excitatory postsynaptic potentials (e.p.s.ps) were recorded from the submandibular parasympathetic ganglia of newborn rats (10-20 days old), by intracellular microelectrode recording and a suction electrode to deliver stimulus trains to the lingual nerve (15 stimuli at 0.1, 0.3, 1, 3, and 10 Hz, 22 degrees C). Only evoked responses without voltage-dependent action potentials were analyzed (observed at membrane potentials negative to -70 mV), and e.p.s.p. amplitudes were determined for the plateau responses during each train (5-15th response). 2. Cadmium, an inorganic calcium channel antagonist, reduced e.p.s.p. amplitudes in a dose-dependent manner (Kd 74 microM, P less than 0.01). Nickel (1 300 microM) did not attenuate the amplitude of evoked responses. 3. Verapamil (0.1-30 microM), a phenylamine, had no significant effects upon e.p.s.p. amplitudes at any frequency examined. Higher concentrations of verapamil (100 microM) blocked neurally evoked responses in a manner consistent with the antagonism of voltage-sensitive sodium currents. 4. Diltiazem, a benzothiazepine, reduced e.p.s.p. amplitudes in a dose-dependent manner, the depression being accentuated at high stimulation frequencies (80% block at 30 microM and 10 Hz). The pure (-)-cis enantiomer of diltiazem (10-30 microM) was without effect. 5. Amlodipine, a 1,4-dihydropyridine, did not antagonize synaptic transmission at any stimulus frequency examined (10-30 microM, 0.1-10 Hz, n = 3). 6. Amiloride, a potassium-sparing diuretic, depressed the amplitudes of evoked responses in a dose-dependent manner (one-site Kd 31 microM, P less than 0.005), although the extent of the block was alleviated with high stimulus frequencies. The effects of 30 microM amiloride were unlikely to be of post-synaptic origin as both the amplitudes of miniature e.p.s.ps, and the iontophoretic potentials induced by exogenous acetylcholine, were not attenuated by treatment with this compound. The amiloride derivative, 3',4'-dichlorobenzamil was ineffective in reducing the amplitude of e.p.s.ps (30-100 microM). 7. omega-Conotoxin GVIA, a marine neurotoxin, which depressed whole cell calcium currents recorded from cultured rat parasympathetic cardiac neurones (up to 90% block at 10 nM), was ineffective at blocking synaptic transmission in submandibular ganglia (0.1-1 microM). 8. The differential effects of these calcium channel antagonists upon synaptic transmission in rat parasympathetic ganglia, suggest that either more than one type of calcium channel may be involved in transmitter release, or that the presynaptic calcium channels possess pharmacological sensitivities different from those of channel types described in ne PMID- 2571383 TI - Sodium salicylate facilitates calcium-dependent release of transmitter at mouse neuromuscular junctions. AB - 1. The effects of sodium salicylate on the frequency of miniature endplate potentials (m.e.p.ps) and on the quantal content of endplate potentials (e.p.ps) in mouse diaphragm muscles at 36 degrees C and 24 degrees C were studied by conventional microelectrode techniques. 2. At 36 degrees C, salicylate (10 mM) elevated the frequency of m.e.p.ps in a manner which was insensitive to [Mg2+]0 and independent of the presence of [Ca2+]0. Lowering the temperature to 24 degrees C abolished the stimulating effect. 2,4-Dinitrophenol (10 microM) had similar effects at 36 degrees C and these disappeared at 24 degrees C. All subsequent experiments were performed at 24 degrees C. 3. Salicylate (10 mM) further increased the frequency (F) of m.e.p.ps stimulated by [Ca2+]0 in a depolarizing solution. When [Ca2+]0 was varied in the absence of salicylate, a linear relationship between ln(F) and ln([ CA2+]0) was obtained. Salicylate shifted this relationship to the left, with respect to the control, without altering the slope. 4. Salicylate (5 mM) also increased the quantal content (m) of e.p.ps in a solution that contained 5 mM Mg2+, in a concentration-dependent fashion. As [Ca2+]0 was varied in the absence of salicylate, a linear relationship between ln(m) and ln[(Ca2+]0) was observed. Salicylate shifted this linear relationship to the left, with respect to the control, without altering the slope. Doubling the concentration of [Mg2+]0 antagonized this effect of salicylate on the quantal content of e.p.ps. 5. These results indicate that salicylate enhances the effect of changing [Ca2+]0. Salicylate probably facilitates the entry of Ca2+ into the nerve terminal or sensitizes the process that is regulated by Ca2 , thereby stimulating the release of transmitter. Surface negative charges may have an important role in the effect of [Ca22]0. PMID- 2571382 TI - Alterations in atrial reactivity in a strain of spontaneously diabetic rats. AB - 1. The present study examined the reactivity of atria from control and spontaneously diabetic rats to various adrenoceptor agonists and to adenosine. 2. Isoprenaline (1.5 nM-1500 nM) produced concentration-dependent increases in inotropy which were unchanged in diabetic atria. However, the sensitivity to isoprenaline-induced changes in chronotropy was reduced in diabetic preparations. 3. In the presence of propranolol (2 microM), phenylephrine (0.2 microM-100 microM) produced concentration-dependent increases in both inotropy and chronotropy; however, atria from diabetic rats exhibited a much greater maximal response. The diabetic state did not alter the sensitivity to phenylephrine. 4. Adenosine (0.15 microM-300 microM) produced concentration-dependent decreases in both inotropy and chronotropy which were unchanged in diabetic atria. 5. Radioligand binding studies revealed that both alpha 1- and beta-adrenoceptor populations were substantially reduced in atria from diabetic rats. However, there was no change in receptor affinity for either adrenoceptor. 6. These results show that diabetes leads to an alteration in atrial reactivity to adrenoceptor stimulation. Future studies examining steps following hormone receptor coupling are required in order to characterize this defect. PMID- 2571384 TI - Comparison of the prejunctional beta-adrenoceptor stimulating actions of adrenaline and isoprenaline in the dog mesenteric vein. AB - 1. The prejunctional beta-adrenoceptor stimulating actions of adrenaline and isoprenaline were compared by recording junction potentials from smooth muscle cells of the dog mesenteric vein. 2. The potency of adrenaline and isoprenaline on beta-adrenoceptors in the postjunctional membrane was estimated from hyperpolarization of the membrane in smooth muscle cells of the guinea-pig facial vein. In the presence of yohimbine, both agents hyperpolarized the membrane to a similar extent. 3. In the dog mesenteric vein, amplitude of the excitatory junction potential (e.j.p.) and slow depolarization was inhibited by adrenaline and potentiated by isoprenaline; the former but not the latter was accompanied by depolarization of the smooth muscle membrane. 4. In the presence of yohimbine, adrenaline inhibited the e.j.p. without depolarization of the smooth muscle membrane; the action was weaker than in the absence of yohimbine. The isoprenaline-induced potentiation of the e.j.p. was further enhanced by yohimbine. 5. It is concluded that adrenaline and isoprenaline have similar stimulating actions on postjunctional beta-adrenoceptors in the guinea-pig facial vein, but have different actions on prejunctional beta-adrenoceptors in the dog mesenteric vein; isoprenaline but not adrenaline stimulates this beta adrenoceptor to facilitate the release of transmitter substances from perivascular noradrenergic nerves. PMID- 2571385 TI - HLA-DO-related restriction fragment length polymorphisms in rheumatoid arthritis: evidence for a link with disease expression. AB - Eighty-three patients with rheumatoid arthritis (RA) were investigated for HLA-DQ and DR locus restriction fragment length polymorphisms (RFLP). Of the 83 patients, 61 (73%) possessed the DR4 allele and within this group we have investigated the relative frequencies of two DQ beta gene variants of DQw3, DQw7 and DQw8, one of which we had previously found to be raised in Felty's syndrome. This analysis revealed a significantly higher frequency of DQw7 containing haplotypes in DR4 positive rheumatoid patients (64%) than in DR-matched healthy controls (42%). Furthermore, the distribution of DQw7 was biased towards those patients with greater disability indicated by the HAQ score, more systemic disease and higher titres of rheumatoid factor, suggesting that DQw7 may contribute to disease expression. PMID- 2571386 TI - Sulphasalazine therapy in ankylosing spondylitis: its effect on disease activity, immunoglobulin A and the complex immunoglobulin A-alpha-1-antitrypsin. AB - Serum levels of immunoglobulin A (IgA) and the complex immunoglobulin A-alpha 1 antitrypsin (IgA-alpha 1AT) were measured at the commencement and after 3 months of a double-blind, placebo-controlled trial of sulphasalazine (SAS) in patients with active ankylosing spondylitis (AS). Twenty-eight patients were evaluated, 15 on sulphasalazine, 13 on placebo. Significant falls were seen in both IgA (p less than 0.01) and IgA-alpha 1AT (p less than 0.001) in the actively treated patients. In addition, significant improvement in clinical and laboratory measures of disease were observed. It is concluded that SAS is effective in AS and modulates the immune response. PMID- 2571387 TI - Haematological side-effects of sulphasalazine in inflammatory arthritis. AB - The nature and incidence of haematological side-effects of sulphasalazine was sought in a retrospective study of 130 sulphasalazine-treated patients with chronic inflammatory arthritis. Macrocytosis was seen to occur in 27 patients (20.8%) and four patients (3%) developed a macrocytic anaemia. Only eight of 23 macrocytic patients had low red cell folate levels, three of whom were anaemic. An increased risk of developing macrocytosis was seen with doses of sulphasalazine greater than 2 g per day. In most patients the macrocytosis was noted during the first 6 months but did occur in the second and third 6-month period of treatment. Only one patient (0.8%) developed neutropenia and no cases of thrombocytopenia were observed. Regular blood counts should be performed while patients remain on treatment but haematological side-effects are seldom the reason for withdrawal of sulphasalazine. PMID- 2571388 TI - The use of sulphasalazine and azathioprine in combination to treat rheumatoid arthritis. PMID- 2571389 TI - Genes and diabetes mellitus. AB - Susceptibility to both insulin-dependent and non-insulin dependent diabetes mellitus is determined to a substantial extent by genetic factors. These probably interact with an environmental trigger to induce expression of the disease state. Significant progress has been made in defining these genetic factors in IDDM. The major contribution comes from a gene or genes within the HLA region, on the short arm of chromosome 6, at least one of which lies close to or within the DQ subregion. Other loci also contribute, most notably the insulin gene, or another closely linked gene on the short arm of chromosome 11, and possibly the T-cell receptor and immunoglobulin genes. In contrast very little is known for NIDDM and such associations as have been described remain controversial. PMID- 2571390 TI - Induction of tyrosine hydroxylase expression in rat forebrain neurons. AB - Olfactory nerve input is required for the normal expression of tyrosine hydroxylase (TH) by dopaminergic neurons in the glomerular region of the rodent main olfactory bulb. To determine whether the olfactory nerve exerts a similar influence on neurons in other brain regions, we performed unilateral bulbectomies in rat pups on postnatal day 5-7 and examined the brains 2-6 months later, after the regenerated olfactory nerve had penetrated the forebrain. Tissue was stained for TH, dopamine beta-hydroxylase (DBH) and olfactory marker protein immunoreactivity. We observed novel TH-immunoreactivity in neurons located in those areas of the adult forebrain which received olfactory nerve fibers, particularly the rostral extension of the subependymal layer. Many of these neurons resembled the periglomerular cells of the olfactory bulb. No cell staining for DBH was observed in these areas, suggesting the possible dopaminergic phenotype of these neurons. Our data indicate that afferent regulation of neurotransmitter expression by the olfactory nerve is not limited to the cells of the olfactory bulb. PMID- 2571391 TI - Dynorphin A-induced rat hindlimb paralysis and spinal cord injury are not altered by the kappa opioid antagonist nor-binaltorphimine. AB - The selective kappa opioid receptor antagonist nor-binaltorphimine (nor-BNI) was used to distinguish a kappa opioid component in the mechanisms underlying the hindlimb paralysis, ischemia, and neuronal injury induced in the rat by the kappa opioid agonist dynorphin A. Spinal intrathecal (i.t.) injection of nor-BNI (20 nmol) either 15 min or immediately before i.t. injections of 5 or 20 nmol of dynorphin A failed to alter the dynorphin A-induced disruption of hindlimb motor function and nociceptive responsiveness. Nor-BNI also did not change the 3-fold increases in cerebrospinal fluid lactate concentrations produced by 20 nmol of dynorphin A. Neuroanatomical evaluations revealed that the cell loss, fiber degeneration, and central gray necrosis in lumbosacral spinal cords of rats treated with 20 nmol of dynorphin A were not altered by nor-BNI (20 nmol, i.t.). Thus, the spinal cord injury and associated neurological deficits resulting from i.t. injection of dynorphin A appear to be primarily, if not totally, attributable to its non-kappa opioid action(s). PMID- 2571392 TI - Glutamate-immunoreactive sensory neurons in quail neural crest cell culture. AB - This study addresses the question of whether embryonic sensory neuroblasts of the quail are glutamate (Glu)-immunoreactive as a neurotransmitter in vivo and when grown in neural crest cell culture. Using a monoclonal antibody against carbodiimide-treated glutamate, we found a subset of neural crest-derived cells in culture. They were large, round and contained long pseudouni- or bipolar processes. There was Glu-immunoreactivity in the cytoplasm of the soma and more intense fluorescence within processes. Double-labeling experiments using a monoclonal antibody to Glu in conjunction with polyclonal antibodies to either neurofilament (NF), substance P (SP), or tyrosine hydroxylase (TH) revealed that: (1) the Glu-immunoreactive cells are a subpopulation of NF-positive sensory neuroblasts; (2) intensely Glu-immunoreactive cells were usually SP-negative. However, cells less intensely staining with both antibodies were observed as well; (3) the Glu-immunoreactive cells were TH-negative, whereas TH-positive adrenergic neuroblasts were Glu-negative; and (4) frozen sections of the spinal ganglia of 9- and 15-day-old quail embryos contained Glu-immunoreactive cells. These data combined indicate that the Glu-immunoreactive cells are a subpopulation of the sensory neuroblasts that were previously identified in neural crest cell cultures with antibodies against NF proteins and the stage specific embryonic antigen 1 (SSEA-1). The results should prove valuable in future studies on the mechanisms that govern neural crest cell differentiation into autonomic and sensory neurons. PMID- 2571393 TI - Glutamate, without GABA antagonists, induces synchronized discharges in intact hippocampus via NMDA receptors. AB - In rats under urethane anesthesia, iontophoresis of high amounts of glutamate (50 150 nA) in hippocampus caused repetitive field potentials. These synchronized discharges were best recorded in the proximal part of stratum radiatum as positive waves of 10-15 ms duration and of 0.5-5 mV amplitude. A tetrodotoxin sensitive faster component of 2-5 ms duration was frequently superimposed on the peaks of the positive waves and was followed by a negative wave of 1-6 mV and 20 30 ms. Glutamate-evoked discharges were suppressed by iontophoresis of N-methyl-D aspartate (NMDA) antagonists, MK-801, Mg2+ and ketamine and also by ketamine injection (i.v. 5-10 mg/kg). The population spikes evoked by fimbrial stimulation were not facilitated by glutamate and the synchronized discharges were suppressed for up to 300 ms following the stimulation, suggesting the presence of an efficient inhibition during glutamate-induced synchronized activity. Glutamate also had no effect on paired-pulse inhibition. No synchronized discharges were recorded with a second electrode separated more than 150 microns from the iontophoretic electrode, suggesting that the activity was local. These data demonstrate that high amounts of glutamate evoke synchronized discharges in hippocampus, possibly through activation of NMDA receptors. The model presented may be utilized to study the mechanisms of synchronization without disinhibition. PMID- 2571394 TI - Glutamate injection into the suprachiasmatic nucleus stimulates brown fat thermogenesis in the rat. AB - Injection of glutamate (100 mM to 1 M, in 0.25 micrograms saline) into the hypothalamic suprachiasmatic nucleus (SCN) dose-dependently increased interscapular brown adipose tissue (IBAT) and core temperatures in the urethane anaesthetized rat. This effect was more pronounced in rats tested during the light-off period than in animals tested during the light-on period. Prior injection of the local anaesthetic, procaine (5% in 0.5 microliter saline), into the ipsilateral ventromedial hypothalamic nucleus (VMH) attenuated the increases in IBAT and core temperatures induced by intra-SCN glutamate. The VMH has previously been implicated in the central regulation of BAT thermogenesis; the present results suggest the pathway arising in the SCN exerts an excitatory influence on VMH neurons involved in the control of BAT function. PMID- 2571395 TI - Bidirectional transfer between kindling induced either by L-glutamate or L aspartate and electrical stimulation in rats. AB - Repeated spaced injections into the amygdala (AM) of rats of a subconvulsive dose of either L-glutamate (GLU) or L-aspartate (ASP) produced progressive seizure development culminating in generalized convulsion strikingly similar to electrically kindled seizure. Both GLU- and ASP-kindled AM sites responded to electrical stimulation with very rapid development of kindled seizure. When electrically kindled AM sites received identical injections of GLU or ASP, a similar positive transfer effect was obtained. These results suggest that (1) either GLU or ASP alone can kindle the AM and (2) GLU, ASP or GLU/ASP kindling share common neurobiological mechanisms with electrical kindling. PMID- 2571396 TI - Radioautographic analysis of somatostatin receptor sub-type in rat hypothalamus. AB - Hypothalamic somatostatin (SRIF) receptors were examined in a qualitative and quantitative radioautographic study using [125I-Tyr0,D-Trp8]SRIF14 and the stable octapeptide analog [125I-Tyr3]SMS 201-995 as radioligands. The latter has been shown to bind selectively to the high-affinity SS1 receptor subtype. Both radioligands labeled specifically and with high resolution various hypothalamic nuclei. In addition, the labeling patterns obtained with the two probes were identical; in both cases specific binding density was highest in the preoptic area and lowest in the ventromedial hypothalamic nucleus. Inhibition of the specific binding of each radioligand by either SRIF14 or the SS1-selective (SMS 201-995) unlabeled competitor was assessed on serial sections throughout the hypothalamus. The proportions of both non-selective and SS1-selective binding, remaining in the presence of either SRIF14 or SMS 201-995 (micromolar concentrations) were identical. These results indicate the existence of a homogeneous class of SRIF binding sites of the SS1 type in the hypothalamus. PMID- 2571397 TI - Glutamine currents in hippocampal neurons are attributable to contaminating glutamate. AB - Physiological responses of cultured rat hippocampal neurons to exogenously applied glutamine were studied using tight-seal, whole-cell voltage clamp techniques in nominal extracellular magnesium. Four mM glutamine induced peak inward currents which were 4.6 +/- 1.7% (+/- 1 S.D.) of current responses to 100 microM glutamate. Ten microM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), 1 mM magnesium, 20 microM MK-801, and 200 microM D-aminophosphonovalerate (APV) blocked these responses in a pattern similar to N-methyl-D-aspartate antagonism. Current-voltage plots produced by 4 mM glutamine and 10 microM glutamate were identical. Even in the absence of extracellular magnesium, 4 mM glutamine was not toxic to our cultured neurons. The data suggest that physiological responses to glutamine are attributable to low concentrations of glutamate in commercial preparations and that glutamine alone has insignificant physiological and toxic effects. PMID- 2571398 TI - Organotypic slice cultures of dopaminergic neurons of substantia nigra. AB - Morphological methods were used to study the plasticity of target-deprived mammalian dopaminergic (DA) neurons. Slices of substantia nigra (SN) were taken from the midbrain of rats aged one to twelve days, and cultured for one to two weeks. Localization of tyrosine hydroxylase (TH) was used to examine the distribution and shapes of DA neurons. Histochemical staining for acetylcholinesterase (AChE) was carried out to estimate both survival and biosynthesis of SN neurons. We found that some DA neurons can survive in vitro without their usual target neurons. This was demonstrated by injecting rhodamine conjugated microspheres (RD) into the caudate putamen, a SN target area, at 6 to 8 days prior to culturing. RD-labeled cells survived in SN cultures and some of them were doubly labeled with AChE. TH neurons had different shapes and their axon terminals formed close contacts with adjacent nondopaminergic neurons. These findings suggest that a subset of DA neurons may switch targets, but the majority of them require target interactions with the caudate putamen for survival in vitro. PMID- 2571400 TI - [Use of recombinant DNA technics in the diagnosis of hereditary diseases]. AB - Recombinant DNA techniques, referred to as genetic engineering or gene manipulation, are used to advantage in medicine for the preparation of hormones, vaccines, immunomodulators, and diagnostic preparations. They enable to introduce new, highly effective diagnostic tools particularly for the identification of defects in the human genome at molecular level. They present methods of direct analysis of genetic material, even without preceding knowledge on the biochemical mechanism of the disease. The aim of the presented paper was to elucidate the substance of direct gene analysis and of indirect analysis by means of genetic linkage, the use of DNA probes and the phenomenon of restriction of fragment length polymorphism (RFLP). The paper gives a concise view on the present state of the problem worldwide as well as on the situation in the Slovak Socialist Republic. (Tab. 1, Fig. 7, Ref. 24.) PMID- 2571399 TI - Direct synaptic contacts of medial septal efferents with somatostatin immunoreactive neurons in the rat hippocampus. AB - Anterogradely labeled projections from the medial septum to hippocampal somatostatin immunoreactive (SOM-i) neurons were studied with double-label immunocytochemistry under light (LM) and electron microscopic (EM) conditions. Medial septal projections were identified after injecting the anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L) followed by immunohistochemical visualization of PHA-L presynaptic terminal labeling and concurrent immunocytochemical staining of SOM in hippocampal target cell bodies. This double label procedure yielded blue-black nickel enhanced DAB stained, PHA-L immunoreactive terminals on light brown SOM-i neurons that were investigated by correlative LM and EM observations. PHA-L-labeled terminal contacts with often basket-like appearance were localized with highest densities on soma and proximal dendrites of SOM-i neurons in stratum oriens of Ammon's horn and hilus of dentate gyrus, and some minor projections to stratum pyramidale and radiatum. Most double labeled contacts could be identified as symmetric type synapses equally divided over soma and proximal dendrites of several forms of SOM-i neurons. These data indicate monosynaptic regulation of the hippocampal intrinsic SOM system by septal input, which probably represents a peptidergic subpopulation of the hippocampal GABAergic system. PMID- 2571401 TI - Cellular resistance to drugs. PMID- 2571402 TI - New antihistamines. PMID- 2571403 TI - Double-blind cross-over trial comparing beclomethasone dipropionate and terfenadine in perennial rhinitis. AB - Eighteen patients with perennial rhinitis were evaluated in this double-blind cross-over trial comparing beclomethasone dipropionate (BDP) aqueous nasal spray with terfenadine tablets. Both treatments were effective in reducing symptom scores but BDP was significantly better than terfenadine in relieving running nose and sneezing (P less than 0.05). BDP also had a greater effect on reducing nasal inflammation than terfenadine. Although the clinicians and patients assessed both therapies to be equi-effective, significantly more patients preferred the BDP treatment (P less than 0.003). Overall, BDP therapy proved more beneficial than terfenadine therapy in this small group of perennial rhinitis sufferers. PMID- 2571404 TI - [Clinical application of a free scapular fascial flap]. AB - The free scapular fascial flap based on the circumflex scapular vessels with skin graft on top has been used to cover the soft-tissue defects in 6 cases--5 hands and 1 foot, over the past one year. The size of the fascial flap ranged from 10 x 6 to 13 x 7 cm. Five flaps were survival completely and one with loss of a small portion due to infection. Compared with the cutaneous flap, myocutaneous flap or muscular flap, the fascial flap is thinner, showing no bulkiness on the recipient site, and also no impairment was noted in the donor site. The shortcoming is darker colour of the skin graft overlying it. The authors consider that the free fascial flap is mainly indicated for the soft-tissue defects of the extremities and those sites where no augmentation is required. PMID- 2571405 TI - Myelinoid bodies in endocrine-paracrine (neuroendocrine, APUD) cells of the prostatourethral region. AB - Distinctive myelinoid bodies (myelinosomes) are described in apparently normal endocrine-paracrine (APUD, neuroendocrine) cells of the prostate and urethra. The ultrastructural findings suggest that this phenomenon represents a form of crinophagy, that is to say autophagocytosis of neuroendocrine granules. PMID- 2571406 TI - Long-range restriction enzyme maps of DNF15S2, D3S2, and c-raf1 loci on the short arm of human chromosome 3. AB - Physical maps have been constructed around loci DNF15S2, D3S2, and c-raf1 on the short arm of human chromosome 3 using pulsed field gradient gel electrophoresis. The normal restriction pattern has not been altered by a t(3;8)(p14.2;q24.1) characteristic for a hereditary form of renal cell carcinoma, indicating that the breakpoint itself is not included in any of the mapped areas. We have found a CpG island within the DNF15S2 locus, suggesting the presence of a functional gene in the region. PMID- 2571407 TI - Enhancement by somatostatin of experimental gastric carcinogenesis induced by N methyl-N'-nitro-N-nitrosoguanidine in Wistar rats. AB - The effects of somatostatin on the incidence, number, and histological type of gastric cancers induced by N-methyl-N'-nitro-N-nitrosoguanidine were investigated in Wistar rats. Rats received alternate-day s.c. injections of 100 or 200 micrograms/kg body weight of somatostatin in depot form after 25 weeks of p.o. treatment with the carcinogen. Prolonged administration of somatostatin at both dosages significantly increased the incidence and number of gastric cancers of the glandular stomach in Week 52. Furthermore, somatostatin at 200 micrograms/kg caused a significant increase in the incidence of gastric cancers penetrating the muscle layer or deeper layers. However, somatostatin at both dosages did not influence their histological appearance. Histologically, somatostatin at both dosages significantly elevated the labeling index of gastric cancers but not of the antral mucosa and significantly reduced the gastrin levels. These findings indicate that somatostatin enhances gastric carcinogenesis after N-methyl-N' nitro-N-nitrosoguanidine treatment and that this effect may be related to its effect in increasing proliferation of gastric cancers and decreasing serum gastrin level. PMID- 2571408 TI - Induction of erythroid differentiation in K562 cells by inhibitors of inosine monophosphate dehydrogenase. AB - The effects of three inhibitors of inosine monophosphate (IMP) dehydrogenase on a human erythroleukemic cell line, K562, were studied. Following incubation with these inhibitors, K562 cells underwent differentiation and accumulated hemoglobins. The induction of hemoglobin accumulation was dose dependent; maximum induction was observed at 100, 25, and 3 microM, respectively, for ribavirin, tiazofurin, and mycophenolic acid. The induction was associated with reduction of intracellular GTP content and was blocked by adding guanosine within 24 h after adding inducer. The effective dose for half-maximum induction by ribavirin was 3 times less than that for 50% inhibition of K562 proliferation; however, for tiazofurin and mycophenolic acid, it closely approximated the concentrations which suppressed cellular proliferation. Ribavirin was sequestered preferentially inside the K562 cells, and the induction by ribavirin had a greater than 30-fold increase in hemoglobin. Studies with isoelectric focusing, globin chain analyses, and immunochemical assays indicated that both A gamma and G gamma were detected and that the hemoglobin produced in the ribavirin-treated cells consisted of approximately 60% fetal hemoglobin and its acetylated equivalents. The adult-type alpha globin was found, while no beta globin chains were demonstrated. Thus, accumulation of fetal hemoglobin and production of alpha globin chain in ribavirin-treated cells are different from the pattern of hemoglobins induced by hemin. PMID- 2571409 TI - Antigens associated with multidrug resistance in H69AR, a small cell lung cancer cell line. AB - In a previous study (S.E.L. Mirski et al., Cancer Res., 47: 2594-2598, 1987), we described the derivation of a multidrug-resistant small cell lung cancer cell line, H69AR. The H69AR cell line does not over-express P-glycoprotein and is therefore a useful model for the investigation of alternate mechanisms of drug resistance. In this paper we report the production and preliminary characterization of six murine monoclonal antibodies (MAbs) which react selectively with the H69AR cell line compared to its drug-sensitive parent cell line, NCI-H69. One of these antibodies, MAb 2.54, detects a cell surface epitope and reacts with multiple proteins of molecular weight 24,500-34,500 on immunoblots. Non-cell surface membrane-associated epitopes are detected by the other five antibodies, MAbs 3.50, 3.80, 3.177, 3.187, and 3.186. MAbs 3.50 and 3.186 immunoprecipitate antigens of molecular weight 55,000 and 36,000, respectively, while MAbs 3.80, 3.177, and 3.187 all precipitate a molecular weight 47,000 protein, suggesting that they may detect epitopes on the same antigen. The epitopes detected by all six antibodies are present on greater than 80% of H69AR cells, as determined by flow cytometry. With the exception of MAb 2.54, the MAbs cross-react in an enzyme-linked immunosorbent assay with the multidrug-resistant human fibrosarcoma cell line HT1080/DR4. Thus, these MAbs react with two drug-resistant cell lines derived from different tumor types in which overexpression of P-glycoprotein is undetectable. These MAbs may detect novel markers for drug resistance and thus may have potential diagnostic or therapeutic value. PMID- 2571410 TI - Screening prescription drugs for possible carcinogenicity: eleven to fifteen years of follow-up. AB - Using computerized pharmacy records from 1969 to 1973 for a cohort of 143,574 members of the Kaiser Permanente Medical Care Program, we have been testing associations of 215 drugs or drug groups with subsequent incidence of cancer at 56 sites. This paper presents findings with follow-up through 1984. There were 227 statistically significant (P less than 0.05, two-tailed) associations: 170 positive, 57 negative. Some were undoubtedly chance findings; others were likely due to confounding by unmeasured covariables. However, several associations suggested hypotheses for further studies and/or the need for continued observation. Most notable among findings not previously reported were associations of several antibiotics, both oral and topical, with lung cancer. These associations could not be explained by indications for drug use or by differences in smoking habits between users and nonusers, and suggest a possible link between the occurrence of bacterial infections and risk for cancer. In general, our results continue to suggest that most medications used during that period did not affect cancer incidence substantially. However, for less frequently prescribed medications, our power to detect moderate increases in cancer risk was quite low. PMID- 2571411 TI - Non-p-glycoprotein-mediated multidrug resistance in detransformed rat cells selected for resistance to methylglyoxal bis(guanylhydrazone). AB - Three independent variants (G2, G4, G5), resistant to methylglyoxal bis(guanylhydrazone), an anticancer drug, have been isolated by single step selection from an adenovirus-transformed rat brain cell line (1). These variants display selective cross-resistance to several natural product drugs of dissimilar structure and action. Multidrug resistance has recently been shown to be caused by overexpression of the membrane-associated p-glycoprotein, most often caused by amplification of the mdr gene. Several types of experiments were conducted to determine whether the observed drug resistance in our cell lines could be due to changes at the mdr locus. The following results were obtained: (a) the mdr locus was not amplified; (b) transcription of the mdr gene and p-glycoprotein synthesis were not increased; (c) multidrug resistance cell lines, which carry an amplified mdr locus, were not cross-resistant to methylglyoxal bis(guanylhydrazone); (d) verapamil did not reverse the resistance of G cells or mdr cells to methylglyoxal bis(guanylhydrazone), nor that of G cells to vincristine; and (e) methylglyoxal bis(guanylhydrazone) resistance was recessive and depended on a block to drug uptake, as opposed to mdr cells which are dominant and express increased drug efflux. The results obtained suggest that the drug resistance in the G2, G4, and G5 cells was atypical and may be due to a mechanism distinct from that mediated by the mdr locus. PMID- 2571413 TI - Gene activity during the early phase of androgen-stimulated rat prostate regrowth. AB - Androgenic steroids regulate the proliferation rate of normal and malignant prostate cells. In order to investigate the molecular basis of this control, we utilized Northern and Western blot techniques to measure changes in the expression of individual genes during the early phase of prostate regrowth. Vestigial ventral prostate glands of 7-day castrated rats showed increased numbers of replicating cells within 12 h of continuous pharmacological testosterone replacement as demonstrated by flow cytometric procedures. The period prior to the onset of proliferative enhancement was accompanied by the sequential induction of a variety of transcripts encoding gene products often associated with cell growth. Within 1 h of treatment, mature mRNA transcripts for c-fos were induced 6-fold, returning to control levels by 2 h. Other genes showed transiently elevated transcript levels after 2 h (c-Ha-ras, c-Ki-ras) or after 8 h (c-myc,c-myb, beta-actin, and Mr 70,000 heat shock protein) of testosterone replacement. Expression of the polypeptide encoded by c-Ha-ras was coordinately enhanced (2-fold) during this period, but not to the levels of the transcript (20 fold induction). Transcripts encoding basic fibroblast growth factor were increased 16 h and later, subsequent to the earlier enhancement in the proliferation rate. By placing these genes in a defined temporal order with regard to their expression in response to testosterone, we have constructed a map of gene activity during early prostate regrowth. This map shows a number of genes, the products of which might participate in the mechanism by which androgens induce mitogenesis of prostate cells. PMID- 2571412 TI - Structure-activity studies of the nonphorbol tumor promoter palytoxin in Swiss 3T3 cells. AB - Derivatives of palytoxin have been prepared which are modified on either the hydroxyl terminus or the amino terminus of the molecule. Previously we have shown that palytoxin, a non-12-O-tetradecanoylphorbol-13-acetate-type tumor promoter, can inhibit epidermal growth factor binding in Swiss 3T3 cells through a pathway which is sodium dependent but not calcium or protein kinase C dependent. We used the epidermal growth factor receptor system to determine whether the specific chemical modifications of palytoxin present in these derivatives alter the cellular mechanism of action of the toxin. The dose response and ion dependence of palytoxin, the hydroxyl terminus derivative palytoxin-COOH, and the amino terminus derivatives N-acetylpalytoxin and N-(p-bromobenzoyl)palytoxin were compared with respect to inhibition of epidermal growth factor binding. The potency of palytoxin-COOH was similar to that of palytoxin. By contrast, N acetylpalytoxin and N-(p-bromobenzoyl)palytoxin were approximately 1/100 as potent as palytoxin in this assay. All three derivatives were at least 100-fold less toxic than palytoxin. Like palytoxin, the activities of palytoxin-COOH, N acetylpalytoxin and N-(p-bromobenzoyl)palytoxin were dependent upon the presence of extracellular sodium. However, there was a significant difference in the dependence of the derivatives on extracellular calcium. Our results suggest that the hydroxyl terminus is important for determining the calcium dependence of the molecule and the amino terminus is important for determining the biological potency of palytoxin. We conclude that modification of the hydroxyl terminus region is an effective means of reducing the toxicity of palytoxin while retaining the biological effects. PMID- 2571414 TI - [Effects of infusions of different doses of dopamine in dilated cardiomyopathy. Observations during and after treatment with beta-blockers]. AB - It is well known that there are abnormalities of the sympathetic nervous system in chronic congestive heart failure. The aim of our study was to verify the effects on heart rate, blood pressure and some echocardiographic parameters of performance and inotropic state of iv infusion of dopamine at different dosages in 14 patients affected by dilated cardiomyopathy. The patients were divided into 3 groups: the first one of patients on standard treatment, the second of subjects on standard treatment and beta blockers, the third of patients who stopped beta blocker therapy and remained on standard therapy. In the first group dopamine at low dosage did not significantly modify heart rate, blood pressure, performance and contractile state, while it did decrease end systolic wall stress. In the second group the same dosage significantly decreased blood pressure and stress, maintaining unchanged the other indexes; the same behaviour was presented by the third group. At high dosage dopamine in the first group significantly increased blood pressure, ejection fraction, contractility and stress. No modifications were observed in the second group, probably because of a pharmacological inhibition of beta receptors. The third group showed a significant increase in blood pressure, stress and inotropic state. From our data it appears that dopamine treatment in chronic congestive heart failure may reveal the presence of the so-called "down regulation" phenomenon. PMID- 2571416 TI - [Perspectives in AIDS chemotherapy]. AB - The author gives a survey of groups of substances which have a marked and selective effect on HI thus serving as potential chemotherapeutical means against AIDS. They can be divided into three groups: 1. anionic substances, 2. 2, 3 dideoxynucleoside analogues and 3. sulphate polysaccharides. The mechanism of the effect of the substances of the 2nd group probably inhibits reverse HIV transcriptase. The substances of the 3rd group are to prevent adsorption of virus particles by the cells. The substances of the 1st group inhibit HIV transcriptase but also interfere with the adsorption process of the virus particles, or with other stages and phases of the HIV replication cycle. The crystallized substances of these groups should be further studied as potential therapeutic agents for the treatment of AIDS and other retrovirus infections (PMEA, D4T, dextran-sulphate, pentosan, polysulphate and others). The survey also includes information on the so-called anti-sense RNA. PMID- 2571415 TI - New therapeutic approaches in rheumatoid arthritis. PMID- 2571417 TI - [Salazosulfapyridine (sulfasalazine)in inflammatory rheumatic diseases]. AB - Salazosulphapyridine is a salicyl derivative of the sulphonamide sulphapyridine. Although its introduction into clinical practice in rheumatology is not recent, only clinical investigations during the last decade proved convincingly its favourable therapeutic effect in rheumatoid arthritis and in some diseases from the group of seronegative spondarthritis. The onset of the therapeutic response after a latency of several weeks, the influence on the laboratory and clinical indicators of the activity of the rheumatic inflammatory response and the concurrent retardation of the development of articular destructive changes on the X-ray picture in some of the patients justify the inclusion of salazosulphapyridine into the group of drugs which modify the course of the disease. The results of hitherto made investigations confirmed also a relatively good tolerance of salazosulphapyridine. PMID- 2571418 TI - Basal body/centriolar DNA: molecular genetic studies in Chlamydomonas. AB - In Chlamydomonas reinhardtii, mutations on an unusual linkage group, the uni linkage group (ULG), affect structure and function of basal bodies. The ULG shows Mendelian segregation, but its genetic map is circular. Molecular cloning of fragments of the ULG was accomplished by taking advantage of restriction fragment length polymorphisms generated by crosses to Chlamydomonas smithii. These clones were used as probes to determine the size and form of the ULG chromosome; it is a 6-9 megabase linear molecule. Use of the probes for in situ DNA hybridization in cells localized the ULG chromosome to basal bodies. PMID- 2571419 TI - The liver-specific transcription factor LF-B1 contains a highly diverged homeobox DNA binding domain. AB - The nuclear protein LF-B1 (also referred to as HNF-1) is a transcription activator required for the expression of several liver-specific genes. LF-B1 has been purified to homogeneity from rat liver nuclear extracts. The sequence of the protein has been partially determined and, subsequently, overlapping cDNA clones containing the entire open reading frame of LF-B1 were isolated. The full-length cDNA encodes a 628 amino acid protein and directs the synthesis in vitro of a protein capable of binding DNA with the same specificity as LF-B1. The cDNA was recombined into a vaccinia virus vector and active LF-B1 was obtained from infected HeLa cells. Addition of the vaccinia recombinant protein to rat spleen extracts results in activation of transcription of an LF-B1-dependent promoter. The DNA binding domain of LF-B1 is located in the amino-terminal part of the protein and displays distant structural similarity to the homeobox domain. The distribution of LF-B1 mRNA is restricted to liver, which correlates with the tissue-specific expression of its target genes. PMID- 2571420 TI - Effects of cyclosporine A on thymocyte differentiation in fetal thymus organ culture. AB - During the course of differentiation in the thymus, precursor T cells are negatively selected by a self-tolerance mechanism or positively selected to acquire restriction specificity to self major histocompatibility complexes. We investigated the process of T cell differentiation and those selections using a fetal thymus organ culture with or without cyclosporine A. The agent blocked the maturation step from CD4+8+ double positive cells to mature CD4-8+ or CD4+8- single positive cells. On the other hand, the agent did not inhibit the development of CD3+4-8- T cell receptor (TCR)alpha beta- cells, which were supposed to be T cells bearing gamma delta-TCR chains. These results suggest that the development of thymocytes bearing alpha beta- or gamma delta-TCR chains differ in requirement for thymocyte-stromal cell interaction. PMID- 2571421 TI - Monoclonal antibodies against LFA-1 or its ligand ICAM-1 accelerate CD2 (T11.1 + T11.2)-mediated T cell proliferation. AB - Activation of human-purified T cells can be mediated by pairwise combinations of monoclonal antibodies directed against T11.1 and T11.2 epitopes on the CD2 molecule. Monoclonal antibodies (mAbs) reactive with either the alpha and beta chains of the lymphocyte-function-associated antigen-1 (LFA-1) molecule or one of its ligands, intercellular adhesion molecule-1 (ICAM-1), were found to accelerate anti-CD2-induced proliferation. This effect was seen on thymocytes and resting or preactivated T cells (phytohemagglutinin blasts and alloproliferative T cell clones) and could be observed, following the introduction of anti-LFA-1 or -ICAM 1 mAbs, up to 50 hr after the CD2 stimulatory signal. This effect was equally abrogated by 55 kDa anti-interleukin-2 (IL-2) receptor mAb, but neither the expression of IL-2 receptor nor the production of IL-2 was modified. The effects of anti-LFA-1 or anti-ICAM-1 on T cell activation through the CD2 pathway were therefore opposite to those observed in the CD3 pathway, where both mAbs strongly delayed T cell proliferation. PMID- 2571422 TI - Genetic interactions in the control of mitosis in fission yeast. PMID- 2571423 TI - Bolus administration of esmolol for the treatment of intraoperative myocardial ischaemia. AB - We report the successful treatment with esmolol of intraoperative myocardial ischaemia associated with concurrent hypertension and tachycardia, in a patient with risk factors for coronary artery disease undergoing peripheral vascular surgery. The pathophysiology of myocardial ischaemia, and the therapeutic role of beta blocking drugs are briefly reviewed. Esmolol, a short-acting cardioselective beta blocking drug, was administered in a bolus of 1.5 mg.kg-1, and resulted in prompt resolution of the haemodynamic abnormalities, with concomitant restitution of the ST segments to isoelectric baseline. We conclude that bolus administration of esmolol is practical and can be effective for the treatment of intraoperative myocardial ischaemia. PMID- 2571424 TI - Should vecuronium be used in renal failure? PMID- 2571425 TI - Comparison of three rat liver foci bioassays--incidence of preneoplastic foci initiated by diethylnitrosamine. AB - Three rat liver foci bioassays have been compared with respect to their sensitivity by the histochemical demonstration of preneoplastic foci, and by the biochemical determination of alterations in enzyme activities of serum indicating hepatotoxicity. We studied the initiation/promotion schedules according to Oesterle and Deml (A), and according to Pereira (B, Broad Spectrum Protocol), and the initiation/selection protocol according to Tatematsu et al. (C), with diethylnitrosamine (DEN), given as a single initiating dose of 10 and 30 mg/kg body wt respectively. With all schedules Sprague-Dawley rats, either females, 3 weeks old (A), or males, 6 weeks old (B, C) were used. For promotion polychlorinated biphenyls (A) or phenobarbital (B) were administered. Selection was performed with 2-acetylaminofluorene (C). The rats in schemes (B) and (C) underwent partial hepatectomy one day prior to initiation. The number and total area of foci deficient in adenosine-5'-triphosphatase (ATPase) and positive in gamma-glutamyltranspeptidase (GGTase) was evaluated. In the complete schedule with 30 mg of DEN in system (A) foci incidence exceeded that of the other systems by about 7-fold (ATPase) and 2-fold (GGTase) respectively. The lower dose of DEN and all control experiments resulted in a respective lower foci yield. With scheme (C), but not with schemes (A) and (B), e.g. serum fructose-1.6 bisphosphatase and alkaline phosphatase were increased, suggesting liver cell damage. Thus tested with DEN, scheme (A) is most sensitive and causes a low impairment of animals' welfare. PMID- 2571426 TI - Glutamine synthetase heterogeneous expression as a marker for the cellular lineage of preneoplastic and neoplastic liver populations. AB - The distribution of glutamine synthetase (GS) in rat liver and in putative preneoplastic and neoplastic lesions was studied at stages of hepatocarcinogenesis induced by diethylnitrosamine (DEN), 2-acetylaminofluorene (AAF) and aflatoxin B1 (AFB1) using immunohistochemistry. In control rats, GS was localized entirely to rings 1-3 cells deep surrounding the central veins. Exposure to DEN or AAF resulted in a reduction of these GS-positive (GS+) zones by 96 and 61% respectively, due to necrosis of the cells in this compartment, while AFB1 had little effect (less than 14%). At later times GS+ foci, adenomas and carcinomas developed in animals exposed to DEN or AAF, but not AFB1, corresponding to the acute effects. Small GS+ foci also identified by other phenotypic abnormalities (e.g. gamma-glutamyltransferase) were exclusively associated with the regenerating GS+ zone as were a few collections of GS+ hepatocytes, the nature of which was uncertain. Although accounting for a very small fraction (less than 3%) of total foci, these GS+ foci or GS+ hepatocytes apparently gave rise to a substantial fraction of adenomas that were GS+ (DEN: 43% and AAF: 33%) and an even higher percentage of GS+ carcinomas (DEN: 59% and AAF: 39%). No GS+ neoplasms were induced by AFB1. Statistical evaluation of these data strongly suggests that GS+ neoplasms originate through initiation of hepatocytes which possess this particular phenotype. Thus, GS might serve as a specific marker for tracing cell lineage relationships during hepatocarcinogensis. The GS+ phenotype which confers glutamine independence may also provide a growth advantage. PMID- 2571427 TI - Possible involvement of arachidonic acid metabolism in phenobarbital promotion of hepatocarcinogenesis. AB - The effects of inhibitors of arachidonic acid metabolism and antioxidants on the rat liver tumor promotion activity of phenobarbital (PB) were assessed using the enzyme-altered focus as the end-point lesion. Fischer 344 male rats were initiated with N-nitrosodiethylamine (200 mg/kg) and then divided into five groups placed on basal diet, diet containing 0.05% PB, diet containing 0.05% PB plus 0.75%, 1% or 1.5% levels of various inhibitors of arachidonic acid metabolism or antioxidants, or diet containing 1% or 1.5% inhibitors or antioxidants alone for 10 weeks, and then killed. p-Bromophenacyl bromide, an inhibitor of phospholipase A2, significantly inhibited the promotion activity of PB at dose levels of 0.75% and 1.5%, reaching plateau at 0.75%. Both quercetin, an inhibitor of lipoxygenase, and morin, a dual inhibitor of lipoxygenase cyclooxygenase, significantly reduced the promotion activity of PB at the 1.5% but not 0.75% dose levels. Moreover, acetylsalicylic acid, an inhibitor of cyclooxygenase dose-dependently inhibited the promotion activity of PB. Among the antioxidants investigated, vitamin E did not affect, but n-propyl gallate and ethoxyquin exerted a dose-dependent inhibition of PB promotion. These results are strongly suggestive of an involvement of phospholipase A2, lipoxygenase and cyclooxygenase arachidonic acid metabolic pathways in the mechanisms underlying PB enhancement of hepatocarcinogenesis. PMID- 2571428 TI - Pharmacologic modulation of increased release of gluconeogenic precursors from extra-splanchnic organs in sepsis. AB - The effect of sterile inflammation and sepsis on the release of lactate and amino acids by peripheral tissues was investigated by removing the splanchnic organs (liver and small intestines) from the circulation and monitoring changes in plasma substrates for 30 min. Functional hepatectomy was performed in rats 5-7 days following the intraperitoneal introduction of a fecal-agar pellet (1.5 ml) [sterile vs. Bacteriodes fragilis (10(8) CFU) + E. coli (10(3) CFU)]. Following functional hepatectomy, dichloroacetate, an activator of the pyruvate dehydrogenase complex, significantly inhibited both lactate and alanine release. L-cycloserine, an inhibitor of alanine aminotransferase, significantly (P less than .05) reduced alanine following hepatectomy. Methionine sulfoximine, an inhibitor of glutamine synthetase, significantly (P less than .005) decreased glutamine accumulation following functional hepatectomy in each of the conditions examined. Treatment with each of these drugs abolished the differences between control and sepsis following hepatectomy. These results demonstrate that alterations in the amino acid profiles during sepsis may be modulated in peripheral organs pharmacologically by utilizing known inhibitors of critical regulatory enzymes. PMID- 2571429 TI - Reperfusion injury and its pharmacologic modification. AB - Reperfusion injury includes a spectrum of events, such as reperfusion arrhythmias, vascular damage and no-reflow, and myocardial functional stunning. The concept of reperfusion injury remains controversial with many proposed mechanisms when applied to humans, whereas in animal models, there are two main proposed mechanisms: calcium over-load and formation of oxygen free radicals. To prove that reperfusion injury is specifically caused by reperfusion would require evidence that an intervention given at the time of reperfusion can diminish or abolish the injury as in the case of arrhythmias, which are thought to be mediated by excess recycling of cytosolic calcium with delayed afterdepolarizations and ventricular automaticity. In the case of myocardial stunning, the phenomenon may be mediated, at least in part, by a burst of free radicals formed within the first minute of reperfusion and improved by free radical scavengers given at the time of reperfusion. The alternate hypothesis is that cytosolic calcium overload damages mechanisms for normal intracellular calcium regulation so that the stunned myocardium responds to agents that are thought to increase intracellular cytosolic calcium, such as beta-receptor agonists. A further component of reperfusion injury, under active investigation, is microvascular damage with alterations at the level of platelets, leukocytes, and endothelial integrity. From the therapeutic point of view, the divergent results of experimental interventions and the possibility that the abrupt onset of reperfusion in animals differs from the situation in humans with thrombolysis means that the best way currently available to limit reperfusion injury is by minimizing the ischemic period by early reperfusion and by optimizing the metabolic status of the ischemic myocardium at the end of the ischemic period. PMID- 2571430 TI - Increased morning incidence of myocardial infarction in the ISAM Study: absence with prior beta-adrenergic blockade. ISAM Study Group. AB - The time of acute myocardial infarction was determined in all 1,741 patients of the ISAM (Intravenous Streptokinase in Acute Myocardial Infarction) Study, based on onset of clinical symptoms and evaluation of plasma CK-MB enzyme time-activity curves. The incidence of myocardial infarction was markedly increased between 6:00 AM and 12:00 noon compared with other times of day (p less than 0.001). Myocardial infarction occurred 3.8 times more frequently between 8:00 and 9:00 AM (hour of maximum incidence) than between 12:00 midnight and 1:00 AM (hour of minimum incidence). Time of myocardial infarction based on clinical and enzymatic methods correlated well (r = 0.95). Patients with higher or lower left ventricular ejection fraction, higher or lower degree of wall motion abnormalities and residual stenosis of the coronary arteries, and one-, two-, or three-vessel disease exhibited a similar circadian pattern, suggesting that the morning is a risk period for patients with mild as well as severe coronary artery disease. Only the group of patients receiving beta-adrenergic blocking therapy before the event did not show an increased morning incidence of myocardial infarction. This observation may contribute to an understanding of the mechanisms by which beta-blockers reduce the incidence of myocardial infarction. Further investigation of physiologic changes occurring during the morning period of increased risk of myocardial infarction may lead to better understanding of the disorder. Design and timing of cardioprotective medication may play a crucial role in improving prevention of acute myocardial infarction. PMID- 2571431 TI - Hemodynamic and beta-adrenergic receptor adaptations during long-term beta adrenoceptor blockade. Studies with acebutolol, atenolol, pindolol, and propranolol in hypertensive patients. AB - In an attempt to further clarify the mechanism of the maintenance of the antihypertensive effect of beta-adrenoceptor antagonists, the effects of four antagonists with different ancillary properties (acebutolol, atenolol, pindolol, and propranolol) on systemic and renal hemodynamics, body fluid volumes, hormones, and lymphocyte beta-adrenoceptor density were studied in four groups of 10 hypertensive patients. The patients were observed for 3 weeks during active treatment and for 2 weeks after withdrawal of treatment. At the end of the 3-week treatment period, the four drugs had an equal antihypertensive effect (fall in mean arterial pressure, 10-13%). Although renin activity was suppressed (60-70%) by all four drugs, changes in renin or pretreatment values of renin levels were not correlated with the fall in blood pressure. The drugs had no effect on plasma catecholamine concentrations or body fluid volumes. Despite similar antihypertensive effects among the four drugs, the changes in flow and resistance underlying the fall in blood pressure differed considerably. With pindolol, the fall in blood pressure was associated with a fall in vascular resistance (26 +/- 6%), whereas with propranolol, it was predominantly associated with a fall in cardiac output (11 +/- 7%). No significant changes in vascular resistance or cardiac output occurred with atenolol or acebutolol. The changes in renal blood flow and renal vascular resistance occurred in parallel with the changes in cardiac output and systemic vascular resistance. Plasma epinephrine concentration and pretreatment cardiac chronotropic responsiveness to isoproterenol appeared to be inversely correlated with lymphocyte beta-adrenoceptor density (Bmax) (r = 0.41 and -0.43, respectively). With pindolol, Bmax decreased maximally by 39 +/- 6%, and with propranolol, it increased by 51 +/- 17%. With both drugs, significant changes in Bmax were already present 24 hours after treatment. Furthermore, 1 week after withdrawal of treatment with pindolol, Bmax was still down-regulated, and cardiac chronotropic responsiveness was still decreased, whereas 1 week after withdrawal of propranolol, Bmax was still up-regulated, and cardiac chronotropic responsiveness was still increased. No changes in Bmax occurred with the beta 1-selective antagonists acebutolol and atenolol. Thus, despite an equal antihypertensive effect, the four beta-adrenoceptor antagonists appear to have dissimilar effects on cardiac output, renal blood flow, and lymphocyte beta-adrenoceptors. Changes in cardiac output, the circulating blood volume, or angiotensin-mediated vasoconstriction are factors unlikely to be crucial for the antihypertensive effect of beta-adrenoceptor antagonists. Therefore, interference with vasoconstrictor nerve activity through blockade of either central or peripheral prejunctional beta-adrenoceptors could be an alternative explanation of their blood pressure-lowering potential. PMID- 2571432 TI - Platelet cytosolic free calcium before and after antihypertensive treatment in perinephritis hypertension of the rabbit. AB - Cytosolic free calcium concentration ([Ca2+]i) in platelets has been reported to be elevated in human essential hypertension, to be positively correlated with blood pressure and to decrease with blood pressure reduction. However, some groups have been unable to confirm these findings in either humans or hypertensive rats. We have examined the relationship between platelet [Ca2+]i and blood pressure in the perinephritis model of hypertension in the rabbit. In addition, the effects of both acute and chronic treatment with verapamil or prazosin were studied. Mean arterial pressure, heart rate and platelet [Ca2+]i were measured before and after treatment. Platelet [Ca2+]i was measured by the Quin 2 fluorescence technique. Platelet [Ca2+]i was similar for the normotensive and hypertensive rabbits, and no correlation between platelet [Ca2+]i and blood pressure was observed. None of the antihypertensive treatments produced a lowering of platelet [Ca2+]i. Therefore we conclude that platelet [Ca2+]i is unlikely to be a universally useful index of ([Ca2+]i in vascular smooth muscle of resistance vessels. PMID- 2571433 TI - Alpha blockers and vasodilating beta blockers--influence on factors involved in the pathogenesis of vascular disease in patients with hypertension. AB - Antihypertensive drugs that interact with adrenoceptors have certain advantages and disadvantages (on the treatment of hypertension). Alpha-1 antagonists such as prazosin have favorable effects on plasma lipids but may produce excessive postural falls in blood pressure, particularly following the initial dose. Recently developed alpha-1 antagonists (doxazosin, terazosin) have longer durations of action than prazosin, allowing less frequent administration. Beta blockers may be cardioprotective but in contrast to alpha-1 antagonists tend to have adverse effects on plasma lipids. Drugs with combined beta and alpha-1 blocking activity such as labetalol have favorable metabolic effects but postural hypotension remains a problem. Recently developed drugs with different alpha 1/beta blocking ratios that differ from labetalol may prove to be more popular clinically. Several beta blockers with vasodilator activity which is not due to alpha-1 blockade have also been developed. These drugs appear to have favorable metabolic effects similar to drugs with alpha-1 blocking activity, but do not cause postural hypotension. PMID- 2571435 TI - 1989 guidelines for the management of mild hypertension: memorandum from a WHO/ISH meeting. PMID- 2571434 TI - Beta-blockers and diuretics. The HAPPHY and MAPHY studies. AB - The question whether initial antihypertensive treatment with a beta-blocker prevents hypertensive complications better than initial treatment with a non betablocker, mainly diuretic based treatment, has been studied in recent large scale studies like the Medical Research Council (MRC) trial, the International Prospective Primary Prevention Study in Hypertension (IPPPSH), the Heart Attack Primary Prevention in Hypertension (HAPPHY) study and the Metoprolol Atherosclerosis Prevention in Hypertensives (MAPHY) study. The first three studies were unable to find a more beneficial effect of beta-blockers than of thiazide diuretics. The MAPHY study, however, found a lower total mortality, in the metoprolol treated group than in the diuretic treated. As the HAPPHY and MAPHY studies share a substantial part of patient-years and number of deaths, these diverging results have evoked confusion and debate. This paper presents the main results of both studies and discusses the possible reasons for the different outcome. PMID- 2571436 TI - Isotype, distribution and target analysis of lymphocyte reactive antibodies in patients with human immunodeficiency virus infection. AB - Anti-lymphocyte (ALA) antibodies were investigated by using both microcytotoxicity and immunofluorescence analyses in 87 subjects with different clinical features of human immunodeficiency virus (HIV) infection. A similar mean percentage of killing in microcytotoxicity assays using heterologous lymphocytes as cellular target was recorded in four groups of patients, including 36 HIV seropositive asymptomatic subjects, 34 patients with HIV-induced lymphadenopathy syndrome (LAS), 13 with acquired immunodeficiency syndrome (AIDS)-related complex (ARC), and 4 patients with the full-blown AIDS. Conversely, an increasing percentage of ALA-positive subjects paralleled the evolution of the HIV infection. The majority of ALA were IgM isotype with a significant reactivity against T cells. This specificity was indifferently directed to CD3+, CD4+, and CD8+ lymphocytes. In additional experiments employing enzymatic digestion of lymphocyte membrane antigens, we demonstrated that CD4 and CD8 receptors were digested by the pronase, whereas CD3 molecules were highly resistant. Subsequent flow cytometry analyses using these pronase-digested T cells showed that reactivity of ALA for their target was unchanged. Our data suggest that antigenic specificities of ALA in HIV infection are resistant to pronase treatment and are not related to CD4 and CD8 molecules. PMID- 2571437 TI - In situ kinetic analysis of thyroid lymphocyte infiltrate in mice developing experimental autoimmune thyroiditis. AB - L3T4+ T cells from genetically susceptible mice developing experimental autoimmune thyroiditis (EAT) were shown earlier to proliferate in response to restimulation with mouse thyroglobulin (MTg) in vitro and to mediate the adoptive transfer of EAT, whereas Lyt-2+ cells differentiated in vitro into cells cytotoxic for thyroid monolayers. Leukocyte suspensions from disrupted thyroid glands examined on Days 13-21 after immunization revealed the accumulation of both T cell subsets in the infiltrate at varying ratios. To characterize the in situ kinetics of cellular infiltration in chronic EAT, we extended the observation intervals after immunization to include Days 21 to 42. The leukocytes in thyroid sections were labeled immunohistochemically first with rat monoclonal antibodies to L3T4, Lyt-2, Thy-1, k light chain, or F4/80 macrophage antigen, then with biotinylated anti-rat IgG, utilizing the avidin-biotin-peroxidase technique. Throughout the 21- to 42-day interval, no significant variations were detected in the percentages of L3T4+ subset, but those of Lyt-2+ cells increased and then declined. The shift in the L3T4+:Lyt-2+ ratio, down from 2.4 to 1.6 and then up to 3.0, was directly related to changes in the Lyt-2+ subpopulation. The F4/80+ and B cell populations changed little during this period. These findings illustrate the changing kinetics of T cell subsets in situ in the development and perpetuation of EAT and MTg-immunized mice. PMID- 2571438 TI - Self-reactive T cells in murine lupus: analysis of genetic contributions and development of self-tolerance. AB - Our understanding of the immune mechanisms that lead to systemic lupus erythematosus has been greatly advanced by the availability of murine models which display both serological and clinical features of the human disease. Studies have demonstrated that CD4+ T cells are required for the full expression of disease in these mice. (NZB X NZW)F1 mice exhibit a lupus-like disease (elevated levels of IgG antinuclear antibodies and a fatal glomerulonephritis) that is not characteristic of either parent. At least three gene loci have been identified in NZW mice that could potentially contribute to a T cell-dependent autoimmune disease, including the T cell receptor alpha- and beta-chain gene complexes and the major histocompatibility complex (MHC). The NZW T cell receptor beta-chain complex appeared to be particularly unusual in that the C beta 1, D beta 2, and J beta 2 gene segments have been deleted. However, an analysis of (NZB X NZW)F1 X NZB back-cross mice revealed no association of disease expression with the presence of this allele. There was also no correlation of disease incidence with the presence of the NZW T cell receptor alpha-chain allele. In contrast, nearly 90% of the backcross mice with the NZW MHC expressed severe autoimmune disease compared with 12% of the mice that did not carry this haplotype. Additional studies strongly suggested that the gene(s) within the NZW MHC is the only dominant NZW genetic contribution to F1 disease. We also determined if self-reactive T cells are able to escape thymic tolerance in autoimmune New Zealand and MRLlpr/lpr mice. In nonautoimmune mice expressing I-E, T cells utilizing V beta 17a and V beta 11 encoded domains have been shown to be clonally eliminated in the thymus. Similarly, V beta 8.1+ and V beta 6+ T cells are tolerized in nonautoimmune mice expressing Mls-1a. These T cell subsets were quantified in the lymph nodes and spleens of (NZB X NZW)F1, (NZB X SWR)F1, and MRL-lpr/lpr mice before and after the development of lupus-like disease. The results indicate that peripheral T cells in these mice, including the massive CD4 , CD8- T cell population in lpr mice, have been modified by normal mechanisms of tolerance such that potential self-reactive V beta specificities have been eliminated in the thymus. PMID- 2571439 TI - Neurohumoral activation in congestive heart failure: a double-edged sword? AB - The search for the basic mechanism(s) responsible for the progressive and frequently irreversible deterioration of left ventricular pump function in congestive heart failure has been quite extensive; nonetheless, no single explanation has been forthcoming. Indeed, given the complexity of this disease process, it is becoming increasingly unlikely that a single pathogenetic mechanism will ever be uncovered for congestive heart failure. This review will examine recent experimental and clinical evidence which suggests that excessive adrenergic stimulation of the heart is double-edged. That is, while increased adrenergic input to the heart may initially enable the failing myocardium to function adequately for a period of months to years, continued excessive adrenergic stimulation of the heart through both local neural and circulating catecholamines may lead to frank myopathic effects on the heart, with resultant worsening of left ventricular function and the development of intractable congestive heart failure. While we do not mean to suggest that excessive sympathetic stimulation of the heart is the only, or even the major mechanism responsible for the development of irreversible congestive heart failure, the data reviewed herein do suggest that adrenergic stimulation may play a primary role in the pathogenesis of congestive heart failure. PMID- 2571440 TI - Ventricular arrhythmias in congestive heart failure. AB - Despite advances in the treatment of congestive heart failure (CHF), the mortality rate continues to be high. A large number of the deaths are sudden, presumably due to ventricular arrhythmias. Complex ventricular arrhythmias are recorded in as many as 80% of patients with CHF, with nonsustained ventricular tachycardia occurring in 40%. The latter appears to be an independent predictor of mortality. Chronic structural abnormalities responsible for CHF may be the basis for the capability of a ventricle to support life-threatening arrhythmias, which are triggered by premature ventricular contractions. The pathogenesis of arrhythmias is multifactorial. Electrolyte abnormalities, ischemia, catecholamines, inotropic and antiarrhythmic drugs may worsen arrhythmias and increase susceptibility of a ventricle to sustained arrhythmias. Beta-adrenergic blockers and angiotensin-converting enzyme inhibitors have a beneficial effect. The role of various drugs in the pathogenesis and treatment of ventricular arrhythmias is discussed. The efficacy of antiarrhythmic therapy targeted to asymptomatic nonsustained ventricular tachycardia, in order to prevent sudden death, is controversial. Pharmacotherapy guided by electrophysiologic testing is the treatment of choice for patients who have manifest sustained ventricular tachycardia, but patients resuscitated from ventricular fibrillation may require automatic implantable cardioverter defibrillator. PMID- 2571441 TI - [Strategy for molecular diagnosis of Duchenne muscular dystrophy: alleles frequency in RFLPs detected with X-linked probes in normal Japanese]. AB - Duchenne muscular dystrophy (DMD) is an X-linked recessive disorder. Recently it has been suggested that the protein named "dystrophin" is not produced in the DMD muscle because of their mutant of defective DMD gene. Recent molecular studies using subclones from the DMD-cDNA as probes revealed a DNA deletion in about a half of the DMD patients. Since not all patients showed deletion, segregation analysis with restriction fragment length polymorphisms (RFLPs) is still necessary and important for diagnosis not only patients but female carriers as well. Therefore, we tried to detect RFLPs with the use of many probes localized within and/or flanked with the DMD gene and made a protocol for the diagnosis with our RFLP data. Among 50 normal Japanese females, the frequencies of rare alleles which identified with enzyme/probe combinations in each were as follows (RFLPs practically available for diagnosis are underlined): 0.38 for p782/EcoRI; 0.14 for p99-6/Pst I: 0.45 for pJ66-HI/Pst I: 0.16 for pERT87-30/Bgl II; 0.44 for 87-15/Xmn I: 0.07 for 87-15/Taq I; 0.33 for 87-8/Taq I; 0.44 for 87-1/Xmn I; 0.45 for 87-1/Mva I; 0.43 for Xj-1.1/Taq I; 0.34 for 754-11/EcoR I; 0.41 for 11.28/Taq I. Some of our results were different from the Caucasian both on the allele frequencies and the restriction fragment length. We proposed a diagnostic protocol of DMD and it's carriers in Japan. Firstly, segregation analysis should be performed with five different combinations of double probe/enzyme, such as Yj 1,.1/L1.28/Taq I, ERT87-1/87-15/Xmn I, 87-1/Mva I, J66-HI/99-6/Pst I, and 782/754 11/EcoR I.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571442 TI - Dosage of intramuscular antipsychotics. PMID- 2571443 TI - Modern treatment of ulcerative colitis. PMID- 2571444 TI - The significance of water absorption and fibre digestion in the omasum of sheep, goats and cattle. AB - 1. The omasal dry matter contents constituted 0.6, 1.2 and 3.3% of the body weight in the sheep, goats and cows, respectively. 2. A large part of the fluid leaving the reticulo-rumen passed quickly through or along the omasum to the abomasum in sheep (43%). The proportion was much less in goats (17%). 3. The absorption of water from the fluid delayed in the omasum was low: 15, 18 and 19% in sheep, goats and cows, respectively. 4. Little or no acid detergent fibre was degraded in the omasum. PMID- 2571445 TI - Iron status, immune capacity and resistance to infections. AB - 1. The importance of iron on immune functions is reviewed. 2. The consequences of iron deficiency upon resistance to infection in men (adults and children) and animals are controversial. 3. Cellular immunity is often altered in iron deficient humans and in murine species. 4. Humoral immune responses seem far less affected in iron-deficient humans than is cellular immunity, but is impaired in iron-deficient animals. Results on complement are scarce and controversial. 5. There is almost no perturbation of phagocytosis but bactericidal activity is decreased in most studies on iron-deficient subjects. 6. Natural Killer activity is decreased in iron-deficient mice. Iron deficiency also affects lymphokine production in mice and rats. PMID- 2571446 TI - Kinetics of D-glucose transport across the intestinal brush-border membrane of the cat. AB - 1. D-glucose transport across the intestinal brush-border membrane of the cat, a carnivorous animal, was investigated using isolated brush-border membrane vesicles (BBMV). Kinetic experiments were performed under zero-trans conditions (initial [Na+]in and [Gluc]in = O) with the transmembrane electrical potential difference clamped to zero. 2. D-glucose uptake by the BBMV was strongly stimulated by an inwardly directed Na+-gradient. Uptake under Na+-free conditions seemed to occur by simple diffusion. 3. The apparent kinetic constants (Vmax, Km) of Na+-dependent D-glucose transport were computed by forcing initial uptake rates at 0.002-10.0 mmol/l D-glucose to either a Michaelis-Menten type equation with a single or with two carrier-mediated components. 4. Best fit of the experimental data was obtained with the two-component model indicating the existence of two Na+-dependent carrier-mediated mechanisms. System 1 and system 2 differ with respect to the transport velocity as well as the substrate affinity constants with Vmax being 2.5-fold and Km being 5-fold higher for system 1 compared with system 2. PMID- 2571447 TI - Chick embryo survival under acute carbon monoxide challenges. AB - 1. Early chick embryos (up to 7 days) survive longer in 1% CO than older ones. As shown by the partition constants of the oxygen-carbon monoxide equilibrium, such a behavior cannot be related to the different hemoglobin patrimonies. 2. In vivo, early embryo hemoglobin is completely turned into its CO derivative within about 15 min according to a step-wise progression which reflects the peculiar embryonic pathway of the blood circulation. 3. Diffusing oxygen is sufficient to keep early embryos alive for a long time. 4. Before the chorioallantois is formed, the vascular area acts as the embryonic respiratory organ. PMID- 2571448 TI - Protein secretion from the cat parotid gland on nerve stimulation. AB - 1. The output and composition of proteins in nerve stimulated saliva samples were compared. 2. Protein output upon parasympathetic stimulation was higher than following sympathetic stimulation and was accompanied by an obvious degranulation of acini with the former but not the latter. These events are the converse of those in the rat parotid gland. 3. Superimposition of sympathetic upon parasympathetic stimulation caused an augmented output of salivary protein. 4. Electrophoresis of salivas revealed differences between individual cats in protein composition but not between differently stimulated salivas. PMID- 2571449 TI - Thermoregulatory responses of diabetic rats. AB - 1. Thermal responses and skin microcirculation were measured in streptozotocin induced diabetic (SD) rats during acute and chronic exposure to ambient (Ta) temperatures ranging from about 5 to 35 degrees C. 2. At 28 degrees C, SD rats had higher rate of oxygen consumptions (VO2), tail skin blood flow (SKBF), but lower rectal temperatures (Tre) than saline-injected controls. 3. Chronic exposure of the SD rats to 35 and 5 degrees C caused a sharp rise and decline in Tre, respectively. 4. At 35 degrees C, hyperthermia in the SD rats was associated with greater increase in VO2 than controls, but changes in SKBF were similar in both groups. 5. At 5 degrees C, VO2 changed similarly in both the SD and control rats, but vasoconstriction was greater in the controls. 6. The data suggest that hypothermia in SD rats may be associated with impairment of vasoconstriction and hyperthermia may be related to an increase VO2 not accompanied by greater vasodilation. PMID- 2571450 TI - The effects of posture changes on blood pressure and heart rate of anesthetized and reserpinized sloths. AB - 1. Tilting sloths anesthetized with chloralose from erect to supine or supine to erect produced little or no effect on heart rate. 2. Tilting anesthetized sloths from erect to supine increased both systolic and diastolic pressures significantly and by about the same amounts. The maximum effect was produced in 20 sec. 3. Pressures stabilized at a higher level than in the erect posture but below the maximum reached in tilting. 4. Tilting these sloths from the supine to the erect posture resulted in a rapid (20 sec) and dramatic fall in pressures to below the initial erect pressure levels. Return to initial erect levels took place slowly. 5. Tilting reserpinized sloths from erect to supine or supine to erect produced little or no effect on heart rate. 6. Tilting reserpinized sloths from erect to supine increased both systolic and diastolic pressures materially and by similar amounts. The maximum effect took 50 sec. 7. Pressures stabilized at higher levels than in the erect posture but less than maximum reached with tilting. 8. Tilting these sloths from supine to erect caused significant falls in pressure to slightly below the initial erect pressure, with maximum effect reached in 30 sec and eventual return to control level. 9. Pressure changes were almost entirely the result of altered venous return. 10. Neither chloralose nor reserpine completely blocked vascular control but reduced it materially. PMID- 2571451 TI - Diphenylamine-2-carboxylate stimulates sodium ion transport in frog skin epithelium. AB - 1. Diphenylamine-2-carboxylate (DPC), added to the mucosal side of the frog skin, increased reversibly the short-circuit current (I0), even in SO2-(4) Ringer. Amiloride blocked this effect. 2. The maximal stimulation was 140% of the control value and the EC50 was 0.26 mM DPC. 3. The stimulatory effect of DPC was additive to that of oxytocin. 4. The dose-response curves for amiloride determined in the absence and in the presence of 1 mM DPC showed an IC50 of 1.0 microM and 0.8 microM amiloride, respectively. 5. Thus DPC, a blocker of Cl- channels in various Cl-transporting epithelia, exerts a stimulatory effect on the amiloride-sensitive Na+ transport in frog skin. PMID- 2571452 TI - Schistosomiasis: role of endogenous opioids in suppression of gonadal steroid secretion. AB - 1. Radioimmunoassay of the opiate, beta-endorphin, in mouse sera, indirect measurement of estrogen by examination of vaginal smears and indirect measurement of androgens by electrophoresis of major urinary proteins (MUP) revealed that beta-endorphin increases while estrogen and androgen levels decrease in mice with chronic Schistosoma mansoni infection. 2. Injections of the opiate antagonist, naltrexone, reversed the effects of schistosomiasis on estrogen and androgen levels. 3. Because opiates are known to inhibit the secretion of releasing hormones by the hypothalamus, the data suggest that the inhibition of hypothalamic-pituitary-gonadal function that occurs in chronically infected male and female mice results from excessive beta-endorphin. 4. It is also suggested that the excessive beta-endorphin may be secreted by T-lymphocytes and possibly macrophages involved in the cell-mediated immune response (CMIR) to the ova. PMID- 2571453 TI - Platelet aggregation in the Asian elephant is not dependent on thromboxane B2 production. AB - 1. The platelet aggregation response to several known platelet agonists was evaluated in four Asian elephants. The platelets were highly responsive to stimulation with platelet-activating factor (PAF) and collagen, less responsive to adenosine diphosphate (ADP) and non-responsive to arachidonic acid, serotonin and epinephrine. 2. Arachidonic acid (1 x 10(-4) M), while inducing no aggregation, caused the release of 1248 +/- 1147 pg/ul (mean +/- SD) of thromboxane B2 (TXB2), the stable metabolite of thromboxane A2 from stimulated platelet. The addition of 1 x 10(-4) M ADP to platelets caused suboptimal aggregation and the release of only 25 +/- 10 pg TXB2/microliters. 3. The calcium channel blocker, verapamil, produced a dose-dependent inhibition of PAF-induced but not collagen-induced aggregation. The cyclooxygenase inhibitor, acetylsalicylic acid, produced no inhibition of either collagen- or PAF-induced aggregation. PMID- 2571454 TI - Calorimetric measurements made on rats during repeated periods of weight gain and weight loss. AB - 1. Rats in respiration chambers were grown for 32 days and their weight reduced by 27-40% on three occasions and realimentated. After the first cycle rats lost body weight more rapidly and regained that weight more quickly than previously. 2. The initial growth phase took 32 days compared with only 8 days in the final period. Each gram of weight gain was associated with 13-14 kJ of dietary net energy during the last two periods of realimentation compared with 19-20 kJ during the first period. 3. Calorimetric measurements showed that although maintenance energy requirement increased during the periods of growth, mean net availability of metabolizable energy was 0.91 compared to 0.60 during weight loss. 4. During the final period much of the weight gain was in the form of lean (67%) but the majority of energy retained was as fat (67-70%). PMID- 2571455 TI - Adaptive behaviour of laboratory rats feeding in hot conditions. AB - 1. A group of six rats, living at thermoneutrality, but with access to food only under a high heat load, was compared with a similar group living and eating at thermoneutrality. 2. Animals eating in the heat reduced total food intake, meal duration, total feeding time, and rate of weight gain; they increased frequency of feeding and had higher water intakes. 3. Identifiable behavioral and nutritional responses explained some of the difference in adaptation between the groups through other factors may also have played a role. PMID- 2571456 TI - Alteration of salt taste sensitivity by the neonatal removal of sublingual glands in the rat. AB - 1. Adult rats with the surgical removal of sublingual glands at their 10 days of age did not prefer NaCl solution of any concentration to water, whereas those with sham-operation or the removal of submandibular glands preferred 0.03 or 0.1 M NaCl to water. 2. Magnitudes of inhibition of chorda tympani responses to NaCl by the lingual treatment of 0.1 mM amiloride were greater in neonatally sublingual removed rats than in sham-operated or submandibular removed ones. 3. These results suggest that the removal of sublingual glands in neonatal period of the rat could increase the amount of the amiloride-sensitive Na receptor mechanism on the taste cell membrane in its adulthood. PMID- 2571457 TI - Relative oxidation of glutamate and glucose by vertebrate erythrocytes. AB - 1. Lungfish erythrocytes (RBC), unlike those in other species of fishes, oxidize endogenous glutamate at a higher rate than they oxidize glucose. This pattern closely resembles that found in invertebrates. 2. The exogenous glutamate oxidation rate of RBC from most species of fish as well as other groups of vertebrates is approximately equal to or less than that observed for glucose. 3. These findings suggest that the nucleated RBC of most vertebrates appear to rely less on the TCA cycle for energy production and more on the glycolytic metabolism of carbohydrates. 4. The RBC of lungfish are also distinguished from RBC of other vertebrates by their relatively higher permeability to exogenous substrates. For example, chicken RBC have a glucose accumulation rate which is approximately one third that observed for lungfish RBC at twice the medium glucose concentration. 5. The unique characteristics of lungfish RBC may be related to their adaptation to the high concentrations of urea produced during estivation. PMID- 2571458 TI - Corticotrigeminal motor pathway in the rat--I. Antidromic activation. AB - 1. The rat corticotrigeminal motor pathway was electrophysiologically investigated. 2. Fifty-one cortical neurons were antidromically activated by stimulation of the contralateral motor trigeminal nucleus (MTN). 3. Twenty-eight of the neurons were examined to see whether they were pyramidal tract (PT) neurons and seven were the PT neurons. 4. Forty peduncular axons were antidromically activated by stimulation of the contralateral MTN and eight of them were the PT axons. 5. Most MTN projecting axons showed slower conduction velocities than their stem anons. PMID- 2571459 TI - Comparative sensitivities of purified preparations of lysyl oxidase and other amine oxidases to active site-directed enzyme inhibitors. AB - Recent evidence has revealed that lysyl oxidase, plasma amine oxidase and diamine oxidase each contain copper and pyrroloquinoline quinone at their active sites as cofactors essential to their catalytic functions. It thus seems likely that these enzymes will share similar mechanisms of action. Since mechanism-based inhibitors of lysyl oxidase have important chemotherapeutic potential for the control of fibrotic disease, the relative inhibitory potential of such agents toward catalytically similar amine oxidases was assessed in the present study using purified preparations of lysyl oxidase, diamine oxidase, plasma amine oxidase and the flavin-dependent mitochondrial monoamine oxidase A and B. The results indicate that there is sufficient difference between the sensitivities of lysyl oxidase and the other amine oxidases to beta-aminopropionitrile to warrant its consideration as an antifibrotic agent in vivo, while also revealing that aminoguanidine, clorgyline and deprenyl are sufficiently selective for diamine oxidase, monoamine oxidase A and monoamide oxidase B, respectively, to differentiate between lysyl oxidase and these enzymes at appropriate concentrations. PMID- 2571461 TI - Current management of acute myocardial infarction. PMID- 2571460 TI - Vecuronium by continuous infusion for neuromuscular blockade in infants and children. AB - Vecuronium's short half-life and minimal cardiopulmonary side-effects make it a suitable drug for continuous infusion. Vecuronium is used frequently in critically ill patients to increase their total compliance and to minimize the adverse effects of mechanical ventilation. This crossover, prospective study evaluates the use of vecuronium by continuous infusion vs. hourly boluses. Patients were assigned randomly to either method; 12 h later each group was transferred to the other method. Neuromuscular blockade (NMB) was followed with the Train-of-Four method. Cardiopulmonary variables were followed hourly. The total dosage/kg body weight was calculated for each method. Six patients were started with continuous drip and the other six with hourly boluses. There were no significant differences in the cardiopulmonary variables through the two periods, although statistically there was a significant difference in the total dosage/kg body weight required for each method. The mean of total vecuronium used in the drip was 0.79 mg/kg.12 h (range 0.1 to 1.8). The mean of the hourly boluses was 1.34 mg/kg.12 h (range 1.0 to 2.55). Patients on continuous infusion required less vecuronium to maintain a similar NMB (p less than .01). PMID- 2571462 TI - Pulmonary capillaritis and hemorrhage. A clue to the diagnosis of systemic necrotizing vasculitis. AB - Patients with systemic necrotizing vasculitis frequently present as diagnostic dilemmas. In previous series of patients with polyarteritis nodosa, less than one third were diagnosed antemortem. Although current clinical awareness of systemic necrotizing vasculitis is greater than in the past and procedures for the diagnosis of these diseases have improved, patients commonly present with atypical features. The diagnosis of a systemic necrotizing vasculitis frequently remains unsuspected or unproven until an involved tissue is biopsied. We report an unusual case of systemic necrotizing vasculitis in which the diagnosis was confirmed by a transbronchial biopsy of the lung which demonstrated pulmonary capillaritis with hemorrhage. PMID- 2571463 TI - The role of oligosaccharides in modifying protein function. AB - It has been proposed that protein-bound oligosaccharides interact with the protein to which they are attached to up- or down-regulate the bioactivity of the 'composite' glycoprotein. Oligosaccharide analyses of the glycoproteins Thy-1, tissue plasminogen activator and immunoglobulin G are presented. Correlations between particular glycoforms and enzymic activities are demonstrated for tissue plasminogen activator. The change in the prevalence of particular immunoglobulin G glycoforms is shown to correlate with disease activity in rheumatoid activity. PMID- 2571464 TI - [Role of electroconvulsive therapy in treating schizophrenia]. AB - Almost half a century has elapsed since the initial application of ECT in psychiatric clinic. It is currently, debatable whether or not ECT in still applicable in schizophrenia, though its effects for manic-depressive disorders is well established. The authors summarized their experiences in evaluating 300 cases, one half use antipsychotic drugs alone and the other half use drugs with ECT it was found that combined therapy favors both groups of patients with aggressiveness or excitement and group with negativism or catatonia as compared in a duration of 2 weeks, whereas patients with hallucinations of suicidal attempts also benefited in 4 weeks' term. On the contrary the combined therapy in the long run is of no value in patients with delusion. PMID- 2571465 TI - [Adams-stokes syndrome due to neuroleptic agents]. AB - Data on an investigation and analysis of 8 cases taken neuroleptics followed by Adams-Stokes syndrome were reported. All patients were diagnosed as schizophrenia without cardiovascular diseases previously. In the process of treatment with 1 or 2 kinds of neuroleptics, Adams-Stokes syndrome appeared, while the potassium was in a range of 1.1--3.8 mEq/L. EKG profile varied, such as showed hypokalemia, ventricular flutter or fibrillation, frequent multifocal ventricular premature beat including bigeminy, and complained with second or third A-V block. The authors deem that neuroleptics may give rise to hypokalemia and then result in cardiac arrhythmias. Damage to heart tends to happen when more than one kind of neuroleptics were medicated. The treatment measures depend above all upon the active and effective attendance to hypokalemia. The patients may die from ventricular fibrillation without proper treatment. PMID- 2571466 TI - [Cardiotoxicity of antipsychotic drugs: abnormal ECG in 1266 cases]. AB - In this report, abnormal changes of 1266 ECG were assessed during antipsyhotic drugs medication. It was founded that cardiac dysrhythmia (sinus tachycardia) in the first place, T wave changes and depression of S-T segment in the next. The rate of abnormal ECG with chlorpromazine seemed to be higher than other drugs. Factors of dosage and time of treatment were non-significant. PMID- 2571468 TI - [Clinical, laboratory and animal experimental observations in treating 288 cases of ischemic cerebrovascular disease with polysaccharide sulfate]. AB - The therapeutic effect and laboratory finding in the treatment of 288 cases of ischemic cerebrovascular disease with PSS were analysed. Positive therapeutic response to PSS in this series of cases was obtained in 92.0% and 62.2% of the treated cases showed excellent results. Effects in the treated patients were better than in the controls. The laboratory findings showed that PSS had obvious anticoagulant effect and decreased blood viscosity and serum contents of lipids. The results of animal experiments showed that PSS had the action of blood dilution, lowering blood viscosity and ameliorating hypercoagulation. PSS was considered to be a prospective, useful drug to prevent and treat ischemic cerebrovascular disease. PMID- 2571467 TI - [An experimental study on the contents of enkephalins in the cerebrospinal fluid in rabbits with unilateral destructive lesions produced in the substantia nigra and their relation to the regulation of MIF-1]. AB - The changes of the contents of enkephalins in cerebrospinal fluid (CSF) and the action of MIF-1 in rabbit experimental models of Parkinson's disease were studied. In the experiment, the rabbits received injection of 6-hydroxydopamine (6-OH DA) into the unilateral substantia nigra. The contents of Met-enkephalin (MEK) and Leuenkephalin (LEK) in the CSF of the fourth ventricles of the normal control rabbits and those with destructive lesions in the substantia nigra were determined with radio-immunoassay. The concentrations of MEK and LEK in CSF of the rabbit models increased markedly to 14.3 and 28.2 folds in the controls respectively. The increased enkephalin content in CSF could be reduced to the normal level by intravenous administration of MIF-I. The results indicated that the action of MIF-I may be one of the important factors in alleviating the symptoms of parkinsonian patients. PMID- 2571469 TI - The use of anxiolytic and parasympathomimetic agents in the treatment of postoperative urinary retention following anorectal surgery. A prospective, randomized, double-blind study. AB - Approximately 30 percent of patients undergoing anorectal surgery will develop acute urinary retention. The cause of this complication is poorly understood. Anxiety, anal distention, bladder distention as a result of vigorous hydration during surgery, and reflex inhibition of the urinary bladder detrusor muscle secondary to pain have been postulated as contributing factors. A four-armed prospective, double-blind, randomized trial was carried out to determine whether an anxiolytic agent (midazolam, 5 mg intramuscularly) and/or a parasympathomimetic agent (bethanechol, 10 mg subcutaneously) reduce the incidence of postoperative urinary retention following anorectal surgery. One hundred thirty-two patients (ages, 18 to 50 years), in acute urinary retention 6 to 12 hours following anorectal surgery, were enrolled. Sixty-nine percent of patients responded to bethanechol. Side effects were minimal. Midazolam alone had no effect on retention. Bethanechol and midazolam in combination resulted in less retention than midazolam and a placebo (P less than 0.05). Bethanechol alone was better than a placebo (P less than 0.002). Mean intraoperative intravenous fluid volume for the entire study group was 900 cc. Initial postoperative urinary volumes of patients who failed the treatment protocol were significantly higher than in those responding to bethanechol (mean of 527 cc vs. 241 cc, P less than 0.001). The use of an anxiolytic agent was not effective in the treatment of postoperative urinary retention. Bladder distention may increase the incidence of urinary retention. Bethanechol, in a dose of 10 mg subcutaneously, significantly lowered the incidence of postoperative urinary catheterization and should be considered as initial treatment of postoperative urinary retention following anorectal surgery. PMID- 2571470 TI - Digestive peptides and asthma. PMID- 2571471 TI - Investigations of the N-hydroxylation of 3'-hydroxyacetanilide, a non-hepatotoxic positional isomer of acetaminophen. AB - The hydroxamic acid of 3'-hydroxyacetanilide (AMAP) was synthesized to test the hypothesis that different reactive metabolites of AMAP and acetaminophen account for similarities in covalent binding of the two positional isomers to hepatic proteins, but for differences in their ability to cause hepatotoxicity. N-OH-AMAP was found to be a relatively stable hydroxamic acid, but it was not detected as a metabolite of AMAP formed in vitro by mouse liver microsomes or in urine of mice administered AMAP. Therefore, metabolites other than N-OH-AMAP must be responsible for covalent binding observed with AMAP to mouse liver proteins. PMID- 2571472 TI - Isozyme-specific monoclonal antibody-directed assessment of induction of hepatic cytochrome p-450 by clotrimazole. AB - Clotrimazole, an N-substituted imidazole widely used as an antifungal agent, has been shown to both inhibit and induce hepatic cytochrome P-450 and related monooxygenase activities. In this study the profile of hepatic cytochrome P-450 isozyme(s) induced by clotrimazole treatment of male Sprague-Dawley rats was investigated. Clotrimazole administration (100 mg/kg, daily for 4 days, ig) resulted in 86% induction of spectrally detectable cytochrome P-450 in hepatic microsomes. In these microsomes 7-ethoxycoumarin O-deethylase (126%), aminopyrine N-demethylase (176%), benzphetamine N-demethylase (117%), p-nitrophenol hydroxylase (89%), and 7-ethoxyresorufin O-deethylase (62%) activities were significantly induced, whereas aryl hydrocarbon hydroxylase activity remained unchanged. Characterization of cytochrome P-450 isozyme(s) in hepatic microsomes prepared from clotrimazole-treated animals was based on the immunoreactivity of these microsomes with highly specific monoclonal antibodies (MAbs) raised against 3-methylcholanthrene-specific P-450 (MAb 1-7-1), phenobarbital-specific P-450 (MAb 2-66-3), pregnenolone-16 alpha-carbonitrile-specific P-450 (MAb C2), and ethanol-inducible P-450 (MAb 1-98-1). Western blot analysis of hepatic microsomes prepared from clotrimazole-treated animals with MAb 2-66-3, MAb 1-98-1, and MAb C2 revealed strong immunoreactive bands, whereas moderate reactivity was observed with MAb 1-7-1. MAb 2-66-3 significantly inhibited 7-ethoxycoumarin O-deethylase activity 45%), whereas MAb 1-7-1 moderately inhibited 7-ethoxyresorufin O deethylase activity (-30%) in clotrimazole-treated animals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571473 TI - Influence of nutritional status on the pharmacokinetics and pharmacodynamics of pentobarbital. AB - The influence of dietary protein deficiency on the pharmacokinetics and pharmacodynamics of pentobarbital was investigated in male Sprague-Dawley rats fed for 4 weeks on a 23% (control) or 5% (low) protein diet ad libitum. Following a single iv dose of 40 mg/kg sodium pentobarbital, the average mean residence time (MRT) was prolonged by 144% (2.3 +/- 0.2 to 5.6 +/- 1.5 hr, mean +/- SD) in the protein-deficient rats, whereas the mean total body clearance (CL) per kilogram of body weight decreased from 0.56 +/- 0.09 to 0.22 +/- 0.06 liter/hr/kg. As a result, the terminal disposition rate constant was decreased by approximately 60% (0.398 +/- 0.037 to 0.178 +/- 0.050 hr-1 when compared to rats on a normal protein diet. No significant differences were found in the two groups of rats with respect to the apparent steady state volume of distribution (Vss). In order to investigate the effect of nutritional status on the concentration pharmacologic activity relationship, pentobarbital was infused iv at a constant rate of 0.55 mg/min until the animals lost their righting reflex (16 +/- 3 min and 8 +/- 1 min in control and protein-deficient animals, respectively). The total dose and concentration of pentobarbital in plasma were not significantly different between the two groups of animals. However, the concentrations of pentobarbital in plasma water (unbound) and brain were appreciably higher in the rats on a low protein diet. Thus, a diet low in protein appears to be associated with a decreased sensitivity of the central nervous system to the depressant effect of pentobarbital. PMID- 2571474 TI - Pharmacokinetics of recombinant human tumor necrosis factor-alpha in rats. Effects of size and number of doses and nephrectomy. AB - The serum pharmacokinetics of recombinant human tumor necrosis factor-alpha (rHuTNF) were determined in male and female rats under various conditions. The clearance of rHuTNF after iv administration was determined to be saturable over the dose range of 10-63 micrograms/kg. Multiple iv administration did not significantly change the clearance of rHuTNF. Nephrectomy significantly reduced but did not eliminate the clearance of rHuTNF. The data suggest that sites other than the kidney also contribute to the elimination of rHuTNF. Female rats showed significantly reduced clearance of rHuTNF compared to male rats at all doses after single and multiple iv administration in control and nephrectomized animals. PMID- 2571475 TI - Pharmacokinetics and excretion of the 21-aminosteroid antioxidant U-74006F in rat and perfused rat liver. AB - U-74006F, 21-(4-(2,6-dipyrrolidinyl-4-pyrimidinyl)-1-piperazinyl)-16 alpha methylpregan-1,4,9(11)-triene monomethane sulfonate, is currently under development for the treatment of human central nervous system trauma and ischemia. The iv pharmacokinetics and excretion of 14CU-74006F (labeled in the 16 alpha-methyl group) and 3HU-74006F (labeled in a pyrrolidine ring) were investigated in the young adult Sprague-Dawley rat and the perfused rat liver. Following a 3 mg/kg iv bolus dose, plasma levels of 14CU-74006F declined biexponentially with alpha and beta half-times of 8 and 70 min, respectively. The terminal phase volume of distribution was 5.1 liters/kg and the plasma clearance was 51 ml/min/kg, which is similar to the in vivo hepatic plasma flow. Plasma levels of total 14C-labeled metabolites quickly exceeded levels of parent drug and declined with a terminal phase half-time of 50 hr. Greater than 90% of the 14C and 3H doses was excreted in feces with terminal phase half-times of 107 and 46 hr, respectively. Consistent with high hepatic clearance, the oral solution bioavailability of U-74006F was 16%, and the hepatic extraction efficiency of U 74006F from 3% bovine serum albumin (w/v) medium in the perfused liver was 80-86% under nonsaturating conditions at physiological flows. U-74006F was rapidly metabolized in the perfused liver and excreted in bile as metabolites. The biliary excretion mechanism was more easily saturated than hepatic uptake and metabolism, with the consequence that, at pharmacologically relevant perfusate levels of drug, metabolites accumulated in the liver and effluxed into the perfusate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571476 TI - The role of flavin-containing monooxygenase in the N-oxidation of the pyrrolizidine alkaloid senecionine. AB - The pyrrolizidine alkaloid, senecionine, is N-oxidized by purified pig liver flavin-containing monooxygenase but not by purified rabbit lung flavin-containing monooxygenase. The activity of the pig liver enzyme toward senecionine was linear with time and amount of enzyme. The oxygenation was not due to some indirect mechanism, such as O2- release from the enzyme, as scavengers of activated oxygen had no effect on product formation. The Km of purified pig liver flavin containing monooxygenase for senecionine was 0.3 mM. Although senecionine is a substrate for the pig liver enzyme, studies performed with rat liver microsomes suggest that, in this species, cytochromes P-450 catalyze the majority of senecionine-N-oxidation. These experiments included inhibition by chemical inhibitors of P-450, treatment of the microsomes with elevated temperatures, inhibition by anti-NADPH-cytochrome P-450 reductase antibody, the effect of dexamethasone on N-oxidation, and relative amounts of flavin-containing monooxygenase determined by immunoquantitation. These results demonstrate that flavin-containing monooxygenase can be involved in the detoxication of pyrrolizidine alkaloids via N-oxidation, but the relative contribution of flavin containing monooxygenase and cytochromes P-450 may be species and tissue dependent. PMID- 2571477 TI - Bioactivation and detoxication of the pyrrolizidine alkaloid senecionine by cytochrome P-450 enzymes in rat liver. AB - Rats display a marked sex difference in the oxidation of the pyrrolizidine alkaloid senecionine, especially with respect to N-oxidation. This sex difference was largely eliminated following treatment with dexamethasone. These observations suggested the potential involvement of the male-specific cytochrome P-450 UT-A and the P-450 PCN-E in the metabolism of this pyrrolizidine alkaloid. Reconstituted rat P-450 UT-A exhibited a high rate of N-oxidation (15 nmol min-1 nmol P-450-1) which is almost 3-fold higher than the turnover number observed with male rat liver microsomes. In contrast, rat P-450 UT-A displayed a much lower activity toward necine pyrrole [+/-)-6,7-dihydro-7-hydroxy-1-hydroxymethyl 5H-pyrrolizine, DHP) formation (1.0 nmol min-1 nmol P-450-1). The N-oxygenation and pyrrole formation activities displayed by rat cytochromes P-450 PB-B and P 450 BNF-B toward senecionine were low, with rates less than 1 nmol min-1 nmol P 450-1. Rabbit antibody to rat P-450 UT-A inhibited the senecionine-N-oxidation activity of untreated male rat liver microsomes by 60%, with lesser inhibition of DHP production. Rabbit antibody to human P-450NF (the human homologue to rat P 450 PCN-E) was a potent inhibitor of DHP production by untreated male rat liver microsomes. With microsomes from dexamethasone-pretreated rats, anti-P-450NF inhibited DHP and N-oxide production in parallel. We conclude that the large sex difference in senecionine N-oxidation probably is the result of the specificity of P-450 isozymes UT-A and PCN-E.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571478 TI - In vitro metabolic degradation of a bovine growth hormone-releasing factor analog Leu27-bGRF(1-29)NH2 in bovine and porcine plasma. Correlation with plasma dipeptidylpeptidase activity. AB - A bovine growth hormone-releasing factor analog, Leu27-bGRF(1-29)NH2, was rapidly hydrolyzed to Leu27-bGRF(3-29)NH2 when incubated at 0.03 mM with porcine and bovine plasma at 37 degrees C in vitro (t1/2 = 8.4 min and 22.1 min, respectively). The site of cleavage was the same as that reported by Frohman et al. (J. Clin. Invest. 78, 906-913, 1986) for the GRF/human plasma system and was suggested by the authors to be due to the presence of dipeptidylpeptidase IV (DPP IV) in human plasma. The DPP-IV-like activity of porcine plasma, determined with Gly-Pro-p-nitroanilide as substrate at pH 7.6 was about 2- to 3-fold higher than that of bovine plasma and seems to correlate well with the more rapid degradation of the GRF analog in porcine plasma. The hormone half-life was extended to 83.3 min when Leu27-bGRF(1-29)NH2 was incubated in vitro with bovine plasma in the presence of an equimolar amount of diprotin A (a competitive DPP-IV inhibitor). Dipeptidylpeptidase II-like activity of porcine and bovine plasma (which may overlap with substrate specificity of DPP-IV) was measured with Lys-Ala-beta naphthylamide and at pH 7.6 was found to be relatively low (3% and 21% of the corresponding plasma DPP-IV activities). Tyr-beta-naphthylamide was hydrolyzed slowly by porcine plasma and not degraded at all by bovine plasma, which suggests that the sequential cleavage from the GRF N-terminus starting with Tyr at position 1 is not dominant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571479 TI - An active, aldehydic metabolite of the cell-differentiating agent hexamethylene bisacetamide. AB - 6-Acetamidohexanal has been identified in human plasma and urine as a metabolite of the cell-differentiating agent, hexamethylene bisacetamide (HMBA). Synthesis of 6-acetamidohexanal was accomplished by oxidation of 6-acetamido-1-hexanol with pyridinium dichromate. A screen of 6-acetamidohexanal for cell-differentiating activity in HL60 human leukemia cells revealed a higher level of activity than HMBA or N-acetyl-1,6-diaminohexane, another active metabolite of HMBA. The results are consistent with a pathway which involves deacetylation of HMBA, oxidative deamination of the resulting N-acetyl-1,6-diaminohexane to yield 6 acetamidohexanal, and further oxidation to give 6-acetamidohexanoic acid. PMID- 2571480 TI - Plasma pharmacokinetics and urinary excretion of thalidomide after oral dosing in healthy male volunteers. AB - The plasma pharmacokinetics and urinary excretion of thalidomide have been evaluated in eight healthy male volunteers receiving a single oral dose of 200 mg. Concentrations of thalidomide were determined by a new HPLC assay. Plasma concentration vs. time data were well fit by a one-compartment model. The mean (+/- SD) peak concentration, 1.15 +/- 0.2 microgram/ml, was achieved at 4.39 +/- 1.27 hr. Absorption and elimination half-lives were 1.70 +/- 1.05 hr and 8.70 +/- 4.11 hr, respectively, with a lag time of 0.41 +/- 0.17 hr observed in six subjects. The apparent volume of distribution and total body clearance rate, based on assumed complete bioavailability, were 120.69 +/- 45.36 liters and 10.41 +/- 2.04 liters/hr. The urinary excretion of thalidomide accounted for only 0.6 +/- 0.22% of the total dose administered over 24 hr, and the renal clearance rate was 0.08 +/- 0.03 liter/hr. This suggests that the major route of elimination of thalidomide is nonrenal. PMID- 2571481 TI - Hydroxylation of 4,4'-methylenebis(2-chloroaniline) by canine, guinea pig, and rat liver microsomes. AB - An in vitro microsomal mixed function oxidase enzyme system was used to study the phase I metabolism of 4,4'-methylenebis(2-chloroaniline) (MBOCA) by dog, guinea pig, and rat liver. TLC with color development and autoradiography, and HPLC with detection by UV absorbance and radioactivity flow monitoring were utilized to isolate metabolites. Reference standards of the N-oxidized metabolites were prepared by oxidation of MBOCA with 3-chloroperoxybenzoic acid and structures confirmed by mass spectrometry and proton NMR. These were utilized to identify the N-hydroxy and nitroso metabolites of MBOCA isolated from the microsomal incubations by comparison of their HPLC retention times and mass spectra. The structure of the o-hydroxy metabolite (ring, ortho to the amine) isolated from the microsomal incubations was elucidated by mass spectrometry and proton NMR. N- and o-hydroxylations of MBOCA were shown to increase with incubation time, microsomal protein, substrate, and NADPH concentration, and were inhibited by 2,3 dichloro-6-phenylphenoxyethylamine, an inhibitor of the microsomal mixed function oxidase enzyme system. Guinea pig liver microsomes oxidized MBOCA to the N hydroxy metabolite predominantly, whereas the dog liver formed predominantly the o-hydroxylated metabolite, with significant amounts of the hydroxylamine as well. The rat liver formed lesser amounts of the N- and o-hydroxylated metabolites, but larger numbers of other polar compounds. PMID- 2571482 TI - Direct determination of diastereomeric carprofen glucuronides in human plasma and urine and preliminary measurements of stereoselective metabolic and renal elimination after oral administration of carprofen in man. AB - A direct stereospecific HPLC assay for carprofen glucuronides in biologic fluids was developed that makes use of a reverse phase (C 18) gradient system, in which the mobile phase consisted of a mixture of acetonitrile and tetrabutylammonium hydroxide buffer (pH 2.5). Reference diastereomeric glucuronides of carprofen were purified from human urine after oral administration of the single enantiomers. When 0.1 ml of sample was used, the limit of detection for carprofen glucuronides was 50 ng/ml in plasma and 200 ng/ml in urine. Coefficients of variation did not exceed 12% for both intra and interday variability. This HPLC method is applicable to pharmacokinetic analysis for carprofen glucuronides in humans. After oral administration of the single enantiomers there was little indication of metabolic inversion. For the two enantiomers the apparent total and metabolic clearances were similar. The limited data available suggest that renal clearance of the (S)-glucuronide was greater than that of the (R)-glucuronide. PMID- 2571483 TI - Disposition of flavodilol in laboratory animals. AB - The disposition of flavodilol, a novel antihypertensive agent, was investigated in rats, rabbits, and dogs following iv or oral administration of 14C-flavodilol or unlabeled drug. Peak, plasma levels occurred within 6 hours of an oral dose in all three species. Following an iv dose, plasma elimination half-lives of flavodilol in rats, rabbits, and dogs were 3.0, 3.0, and 4.0 hr, respectively. Total body clearances were 0.71 liter/hr/kg for the rat, 1.89 liters/hr/kg for the rabbit, and 3.07 liters/hr/kg for the dog. Renal clearances were a small fraction of total clearance at 0.042, and 0.114 liter/hr/kg for the rat and dog, respectively, suggesting extensive nonrenal clearance. The volumes of distribution of 3.04 for the rat, 8.10 for rabbit, and 18.13 liters/kg for dog are large, suggesting significant extravascular distribution of flavodilol. Following 10 and 50 mg/kg po doses of 14C-flavodilol in rats, recovery of total radioactivity after 79 hr was 100.7% and 88.4% of the dose, respectively, most of which was recovered in the feces (77.5% and 66.6%, respectively). Tissue distribution studies of 14C in rats at 1.5, 5, 24, and 48 hr after a single po dose of 10 mg/kg 14C-flavodilol showed that the majority of the radioactivity was in the gastrointestinal tract and organs of elimination at all time points. Less than 1% of the dose remained in the body at 48 hr. 14C-Flavodilol was administered to rats iv at 1 mg/kg and orally at 10 mg/kg to assess comparative (label vs. nonlabel) absorption and distribution characteristics.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571484 TI - Dose-dependent kinetics of cilastatin in laboratory animals. AB - Cilastatin, a potent inhibitor of renal dehydropeptidase I, was specifically designed to inhibit renal metabolism of the antibiotic imipenem in order to achieve therapeutically relevant imipenem concentrations in the urinary tract. In this study the elimination kinetics of cilastatin in rats at doses of 5, 10, 20, 50, 100, and 200 mg/kg iv were demonstrated to be dose dependent, with total plasma clearance and non-renal clearance falling from 20.2 +/- 3.1 ml/min/kg and 17.7 +/- 3.3 ml/min/kg (mean +/- S.D.) at the 5 mg/kg dose to 11.4 +/- 1.2 ml/min/kg and 5.30 +/- 1.2 ml/min/kg, respectively, at the 200 mg/kg dose, whereas the volume of distribution of the drug remained unchanged. Since cilastatin is mainly eliminated by renal excretion as well as by N-acetylation, the non-renal clearance may reasonably reflect the N-acetylation process. Thus, the dose-dependent kinetics of cilastatin might be explained, at least partly, by the saturation of the N-acetylation of the drug. The dose-related decrease in the fraction (fm) of cilastatin converted to its N-acetylated metabolite provided further evidence for the saturable N-acetylation. The fm values decreased from 0.915 at the 10 mg/kg dose to 0.626 at the 100 mg/kg dose. Although both the total plasma clearance and non-renal clearance decreased with increasing dose, the dose had an opposite effect on the renal clearance of cilastatin. The renal clearance of cilastatin increased from 2.50 +/- 0.40 ml/min/kg at the lowest dose to 6.10 +/- 0.50 ml/min/kg at the highest dose as the dose increased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571485 TI - Glucuronidation of E-10-hydroxynortriptyline in human liver, kidney, and intestine. Organ-specific differences in enantioselectivity. PMID- 2571486 TI - Doxofylline in rat brain in relation to locomotor activity. AB - Doxofylline is a new xanthine derivative with significant bronchodilatatory activity. We have studied in HPLC the distribution of doxofylline in various areas of rat brain (cortex, cerebellum, limbic system) and its activity on the central nervous system by using a spontaneous motility test comparing it with aminophylline administered orally in equimolar doses (4.7 - 9.4 - 19 x 10(-5) mol/kg). Doxofylline is absorbed 3 or 4 times less than aminophylline at the same doses. Nevertheless, the quantity of doxofylline that goes to the brain is equivalent to that absorbed, whereas the quantity of aminophylline is about one third. This is due to the greater liposolubility of doxofylline in comparison to aminophylline. In spite of the fact that doxofylline is easily distributed in the brain, spontaneous motility in animals is not modified, whereas aminophylline increases this activity significantly. The low affinity of doxofylline with adenosine receptors (A1 and A2) in comparison with aminophylline explains the lack of side effects on the central nervous system which has been amply documented for theophylline and for other methylxanthine derivatives. PMID- 2571487 TI - 3,3',4,4'-Tetrachlorobiphenyl. Excretion and tissue retention of hydroxylated metabolites in the mouse. AB - The coplanar 3,3',4,4'-tetrachlorobiphenyl (TCB) was given orally to mice and the metabolite patterns in feces, urine, liver, and adipose tissue were examined. In feces, 80% of the dose was excreted within 5 days. 5-Hydroxy-, 6-hydroxy-TCB, 4 hydroxy-3,3',4',5-tetrachlorobiphenyl, and unmetabolized TCB were identified by comparison to synthetic standards (GC/MS). 4-Hydroxy-trichlorobiphenyl and a dihydroxy-trichlorobiphenyl were indicated by the fragmentation pattern of the corresponding methylated derivatives by GC/MS. In urine, 4.9% of the TCB dose was excreted mainly as conjugates. After hydrolysis, TCB and seven hydroxylated metabolites were detected; 2-, 5-, and 6-hydroxy-TCB and 4-hydroxy-3,3',4',5 tetrachlorobiphenyl were identified and two dihydroxy-tetrachlorobiphenyls were indicated. The major compound detected after hydrolysis of urine was a dihydroxy trichlorobiphenyl. TCB was the major compound present in the liver, while a minor portion was due to 4-hydroxy-3,3',4',5-tetrachlorobiphenyl. TCB, 4-hydroxy 3,3',4',5-tetrachlorobiphenyl, and 5- and 6-hydroxy-TCB were present in adipose tissue. In addition, radiolabeled material was present in a lipid fraction obtained after gel permeation chromatography of all samples except urine, indicating the presence of TCB metabolites with lipid characteristics. PMID- 2571488 TI - In vitro formation of an inhibitory complex between an isosafrole metabolite and rat hepatic cytochrome P-450 PB-B. AB - A series of in vitro studies was performed, in rat liver microsomes, in which metabolite intermediate (MI) complexation of cytochrome P-450 (P-450) by the methylenedioxyphenyl compound isosafrole was related to P-450 isozyme-specific inhibition of drug oxidation. The C19-steroid androst-4-ene-3,17-dione was selected for initial study because the stereoselective hydroxylation of this substrate is specific for certain P-450s. In control microsomes only the 6 beta- and 16 beta-hydroxylations of the steroid (catalyzed, respectively, by the P-450s PCN-E and PB-B) were inhibited by isosafrole (I50 = 100 and 110 microM). In contrast, the 7 alpha- and 16 alpha-hydroxylases (P-450 UT-F- and UT-A-mediated, respectively) were refractory to inhibition. After phenobarbital (PB) induction, steroid 6 beta- and 16 beta-hydroxylase activities were again inhibited (I50 = 170 and 190 microM) but, in addition, the 16 alpha-hydroxylase pathway was also inhibited (I50 = 200 microM). Spectral studies revealed that MI complexation of P 450 in untreated microsomes was minimal but was enhanced markedly after PB induction (up to 50% of the total P-450 content complexed). Thus, it is apparent that a PB-inducible P-450 is involved in MI complex formation under these conditions. Indeed the I50 of isosafrole toward steroid 16 beta-hydroxylase activity was decreased if the inhibitor was preincubated with NADPH-fortified PB induced microsomes prior to substrate addition; the preincubation step did not enhance the inhibition of any other steroid hydroxylase pathway by isosafrole.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571489 TI - Studies of metabolic pathways of trimebutine by simultaneous administration of trimebutine and its deuterium-labeled metabolite. AB - Trimebutine maleate (I), (+-)-2-dimethylamino-2-phenylbutyl 3,4,5 trimethoxybenzoate hydrogen maleate, and a deuterium-labeled sample of its hydrolyzed metabolite, 2-dimethylamino-2-phenylbutanol-d3 (II-d3), were simultaneously administered to experimental animals at an oral dose of 10 or 50 mumol/kg, and distribution ratios of the two alternative initial metabolic steps, i.e., ester hydrolysis and N-demethylation, were estimated by determining the composition of the urinary alcohol-moiety metabolites, II, and its mono- and di demethylated metabolites, III and IV, by GC/MS. In dogs, the order of quantities of the metabolites from II-d3 was II much greater than III much greater than IV, showing predominance of conjugation over N-demethylation. However, this order was reversed when the amounts of the metabolites from I were compared, indicating that I was preferentially metabolized by N-demethylation followed by ester hydrolysis and conjugation in this order. In rats, a considerable proportion of I was presumed to be metabolized by ester hydrolysis before N-demethylation. In in vitro experiments employing the liver microsomes and homogenates of liver and small intestine from rats and dogs, it was found that both ester-hydrolizing and N-demethylating activities were higher in rats than in dogs, and the conjugating activity was higher in dogs than in rats. It was also found that I, having a high lipophilicity, was more susceptible to N-demethylation than less lipophilic II. These results from the in vitro experiments could account for the species differences in the distribution ratio of the metabolic pathways of I in vivo. PMID- 2571490 TI - Oxidation of tenoxicam by leukocyte peroxidases and H2O2 produces novel products. AB - A leukocyte extract, which had a high peroxidase activity (mostly myeloperoxidase), converted tenoxicam [4-hydroxy-N-(2'-pyridyl)-2-methyl-2H thieno-(2,3e)-1,2-thiazine-3 - carboxamide-1,1-dioxide] a potent antiinflammatory drug, into four novel metabolites in the presence of H2O2: 4,5-dihydro-4-oxo-5 methyliminopyrido (1,2a) imidazole (metabolite I), 2-carboxyl-3-thiofenesulfinic acid (metabolite II), 2-carboxyl-3-thiofenesulfonic acid (metabolite III), and N methyl-N'-(2-pyridyl)oxamide (metabolite IV). These metabolites were probably formed by a one-electron oxidation reaction at the center carbon atom of the beta diketone moiety of tenoxicam. Tenoxicam is a cofactor for the reduction of peroxidases and this capability may explain at least a part of the antiinflammatory effect of tenoxicam. PMID- 2571491 TI - The identification and management of drug-seeking behavior in a medical center. AB - We describe the development of a quality assurance program that monitors prescription medication misuse in a medical setting. The program focuses on patient activities that influence physician prescribing of abusable medications. Seven defining criteria have been developed to judge the presence of drug seeking by patients. When appropriate, a drug-seeking label is attached to the patient's chart and the hospital computer information system. The warning informs the physician and protects the patient from excessive medications. PMID- 2571493 TI - [Polymorphism of restriction fragments of the Ha-ras-1 protooncogene in patients with carcinoma and ulcers of the stomach]. AB - Ha-ras restriction fragments' length polymorphism (RFLP) in white blood cells and stomach tissues from patients with carcinoma and ulcer of the stomach was examined. Genomic DNAs were digested with Xho I, Pvu II, Pst I, Msp I, Bcn I, Mva I, Bsp RI. No Ha-ras-1 polymorphic variants specifically associated with the cancer disease were detected. RFLP of the 3'-noncoding sequence of c-Ha-ras-1 gene had got features of the definite human population. The cells forming the tissue were examined and individual peculiarity of the methylated residues disposition seemed to influence RFLP of the 5'-noncoding sequence of c-Ha-ras-1. PMID- 2571492 TI - Lefetamine: new abuse of an old drug--clinical evaluation of opioid activity. AB - Lefetamine (SPA) combining amphetamine with opioid-like effects, a drug of wide abuse in Japan in the fifties, has now been introduced as such in Italy. In this study the drug was tested to verify its resemblance to opiates. Ten lefetamine abusers were hospitalized and then subjected to naloxone- and pentazocine-tests and detoxified. Moreover, lefetamine was administered to ten opiate addicts with an acute withdrawal syndrome and to ten methadone-treated addicts. The naloxone test was positive and pentazocine could be substituted for lefetamine. Lefetamine was able to relieve opiate withdrawal and did not precipitate withdrawal symptoms in stabilized opiate addicts. It is concluded that lefetamine may act as an opioid partial agonist. PMID- 2571496 TI - Percutaneous transhepatic cholecysto-lithotripsy (PTCL). AB - Percutaneous transhepatic cholecystolithotripsy (PTCL) by laser or ultrasound is a minimally invasive form of treatment for gallstones. All the steps of the procedure can be performed in a single session. In clinical use severe side effects have not been observed. PMID- 2571494 TI - Sexually dimorphic characteristics of clonidine-induced growth hormone release and autofeedback. AB - The mechanism underlying the sexually dimorphic pattern of GH secretion in the rat has not been clearly elucidated. In this study differences in the regulation of GH secretion in males and females were analyzed by examining their sensitivity to alpha 2-adrenergic (alpha 2) stimulation and GH autofeedback. The model used in testing this system was the capacity of ovine (o) GH, injected into the third ventricle, to suppress the endogenous GH surge induced by clonidine (CLON), an alpha 2-agonist. In morning experiments, CLON (30 micrograms/kg BW, iv) was injected between 1000-1100 h in males. oGH (20 micrograms in 2 microliters vehicle) injected 3 h earlier inhibited the GH surge, which peaked 15 min after CLON injection in control animals receiving CLON alone. In females there was both no change in plasma GH levels and no difference between treatment groups in animals receiving 30, 50, 100, or 150 micrograms/kg CLON and saline controls. Preinjection of 20 micrograms oGH in proestrous or diestrous animals or 30 micrograms oGH in diestrous animals 3 h before CLON treatment did not depress plasma GH levels below those found in animals receiving CLON alone. Further, neither reducing the preinjection time to 2 h nor injecting 7, 13, or 20 micrograms oGH at this preinjection period suppressed basal or CLON-induced GH release. To facilitate comparisons in subsequent experiments the male protocol was followed. In evening experiments, CLON (injected between 2210-2240 h) stimulated a significant GH surge in diestrous females. However, preinjected oGH lowered basal GH levels but failed to suppress the CLON-induced GH surge. In somatostatin (SRIF) antiserum (anti-SRIF; 0.5 ml, iv)-treated adult diestrous animals, CLON induced a significant GH surge that was suppressed by oGH preinjection. In normal sheep serum-treated animals there was no significant response to either CLON alone or in combination with oGH. To determine the influence of ovarian hormones on GH release females were ovariectomized (OVXed) either pre- or postpubertally. After adult OVX, CLON induced an increase in GH release that was not suppressed by oGH pretreatment. In prepubertally OVXed animals CLON induced a substantial GH surge that was suppressed in animals preinjected with oGH. In prepubertally OVXed animals implanted with testosterone capsules the response to CLON and oGH was not significantly different from that after prepubertal OVX alone. Sham-operated animals responded to treatment in a manner similar to unoperated cycling females.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2571495 TI - Alpha 1- and beta-adrenergic agents cause synergistic stimulation of the iodothyronine deiodinase in rat brown adipocytes. AB - Previous studies have shown that norepinephrine causes a marked increase in the iodothyronine 5'-deiodinase activity in dispersed brown adipocytes. This stimulation required mRNA and protein synthesis and was 3- to 4-fold greater in cells from hypothyroid than in those from euthyroid rats. To investigate the cause of this differential response, we incubated dispersed brown adipocytes with catecholamines, a specific alpha 1-agonist, forskolin, and (Bu)2cAMP alone and in combination. Our results show a synergistic effect of alpha 1- and beta adrenergic catecholamines to increase the deiodinase, which leads to 2-fold (euthyroid) to 4-fold (hypothyroid) higher enzyme activities in the presence of both agonists than can be accounted for by additive effects of the two agents. Since alpha 1-agonists cause minimal stimulation alone, this response is due to an enhancement of the cAMP effect. The alpha 1 effect is mimicked by the calcium ionophore A23187, but not by phorbol ester alone. After 2-h exposure to beta adrenergic agents or forskolin, hypothyroid cells had a reduced cAMP response, but alpha 1-agonists did not reverse this. These results demonstrate a complex interrelationship between alpha 1- and beta-adrenergic agonists and thyroid status in the regulation of deiodinase in the brown adipocyte. The increase in intracellular calcium due to an alpha 1-agonist markedly enhances the effects of cAMP on deiodinase activation, permitting a beta-adrenergic effect despite the impaired cAMP generation characteristic of hypothyroid adipocytes. This unexpected enhancement of the beta-adrenergic pathway in the hypothyroid state may be especially relevant for maintaining maximum T3 production, which is required for the normal thermogenic function of this tissue. PMID- 2571497 TI - Treatment of infantile spasms with methysergide and alpha-methylparatyrosine. AB - Twenty-four newly diagnosed and previously untreated infantile spasm patients were treated for 3 weeks with either methysergide (12 patients) or alpha methylparatyrosine (12 patients). Response to therapy was determined objectively with 24-h polygraphic/video monitoring techniques and was defined as cessation of spasms and disappearance of the hypsarrhythmic EEG pattern. Two (17%) of the patients treated with alpha-methylparatyrosine responded to therapy, and one (8%) of the methysergide-treated group showed a response. PMID- 2571498 TI - Effect of acute and chronic terfenadine on free and total serum phenytoin concentrations in epileptic patients. AB - Terfenadine is a unique H1-receptor antagonist devoid of adverse central nervous system (CNS) effects. Terfenadine is highly protein bound and has been shown to stimulate hepatic microsomal enzymes, making an interaction with phenytoin (PHT) possible. After assuring constant PHT levels for 7 days, 12 epileptic patients were studied with PHT alone, after a single daily dose of terfenadine (60 mg twice daily), and again after 2 weeks of chronic terfenadine therapy. Samples for PHT were drawn over 24 h, and urine was collected for determination of the PHT metabolite 5-(p-hydroxyphenyl)-5-phenylhydantoin (HPPH). The pharmacokinetic parameters calculated for total and free PHT and HPPH were area under the time concentration curve (AUC), the maximum serum concentration (Cmax), the minimum serum concentration (Cmin), and the percentage of fluctuation between maximum and minimum concentrations. A factor analysis was used to clarify interrelationships. Five dependent variables were found to represent the data. These were assessed by multi-variate analysis of variance (MANOVA); p less than 0.05 was considered significant. No significant differences were observed for any of the parameters for the acute or chronic effects of terfenadine. We conclude that terfenadine does not interfere with PHT. PMID- 2571499 TI - The gene coding for the major birch pollen allergen Betv1, is highly homologous to a pea disease resistance response gene. AB - Pollen of the white birch (Betula verrucosa) is one of the main causes of Type I allergic reactions (allergic rhinoconjunctivitis, allergic bronchial asthma) in Middle and Northern Europe, North America and the USSR. Type I allergies are a major threat to public health in these countries, since 10-15% of the population suffer from these diseases. BetvI, an allergenic protein with an Mr of 17 kd is a constituent of the pollen of white birch and is responsible for IgE binding in more than 95% of birch pollen allergic patients. Here, we report the complete nucleotide sequence and deduced amino acid sequence of a cDNA clone coding for the major pollen allergen (BetvI) of white birch. It is similar to the N-terminal peptide sequences of the allergens of hazel, alder and hornbeam (close relatives) but it has no significant sequence homology to any other known allergens. However, it shows 55% sequence identity with a pea disease resistance response gene, indicating that BetvI may be involved in pathogen resistance of pollen. PMID- 2571501 TI - Onset and duration of clinical relaxation of atracurium and vecuronium in patients on chronic nifedipine therapy. AB - The onset and duration of clinical relaxation of atracurium and vecuronium were studied in patients on chronic nifedipine therapy and compared to similar groups of patients not on such a therapy. Neuromuscular block was monitored by recording the integrated electromyogram of the hypothenar muscles (using a 'Datex Relaxograph') following stimulation of the ulnar nerve with a train-of-four stimulation. Neither the time to onset of maximum block (3.0-3.4 min) nor the duration of clinical relaxation (36-40 min) differed significantly between the groups indicating no significant interaction in humans between atracurium and vecuronium and chronic nifedipine therapy. PMID- 2571502 TI - Saccadic reaction times in acute and remitted schizophrenics. AB - Schizophrenics are known to have various disturbances of the visuomotor system. Whereas smooth pursuit eye movement disorders have been repeatedly confirmed, there are relatively few reports regarding possible disturbances of the saccadic system. In this study, the saccadic reaction times of 47 schizophrenic inpatients were investigated upon admission and later in the remitted state; 28 age- and sex matched normal volunteers were tested as controls. Psychopathology and outcome were evaluated according to the Brief Psychiatric Rating Scale and the Prognostic Scale. Light stimuli were presented at random direction, location (ranging from 0 degrees to 20 degrees) and duration (800, 1000, and 1200 ms). The eye movements were recorded by electro-oculography. Compared with the control group, schizophrenics revealed prolonged saccadic reaction times, which correlated with pronounced negative symptoms and an unfavourable course of the illness. The saccadic reaction times remained prolonged in schizophrenic patients. These findings suggest attentional deficits in schizophrenics. PMID- 2571500 TI - Association of two pertussis toxin-sensitive G-proteins with the D2-dopamine receptor from bovine striatum. AB - The solubilized D2-dopamine receptor from bovine striatum exhibits high and low affinity states for dopaminergic agonists. Guanine nucleotides and pertussis toxin convert the solubilized receptor from a high affinity state to a low one. A D2-receptor preparation partially purified by affinity chromatography on a haloperidol adsorbent, exhibited agonist-stimulated GTPase activity. [32P]ADP ribosylation by pertussis toxin of this receptor preparation resulted in the specific labeling of two protein bands corresponding to mol. wts of 39 and 41 kd, in SDS-PAGE. Association of these G-proteins with the receptor was specifically inhibited by Gpp(NH)p. Immunoblot analysis of these G-proteins indicated that the 41- and 39-kd protein bands are analogous to brain Gi and Go respectively. These experiments demonstrate that two distinct pertussis toxin-sensitive G-proteins are functionally associated with bovine striatum D2-dopamine receptor. PMID- 2571503 TI - Impaired performance in a saccadic tracking task in schizophrenic patients. AB - The performance of schizophrenic patients in a task which requires a sequence of saccades guided by visuospatial cues is reported. A previous study recording eye movements determined a highly increased number of fixations necessary for this task in acute schizophrenic patients compared with normal controls. The performance of normal control subjects of different age groups and the correlation between the performance in the tracking task and the results of a clinical psychological test battery are described. Schizophrenic patients in a partly remitted state and in a remitted or mildly chronic state performed this task worse than matched control subjects; this was particularly indicated by the time score. The relation to the lifetime dosage of neuroleptic medication is considered. PMID- 2571504 TI - Utilization of benzodiazepines in Chile during 1982-1986. AB - The consumption of benzodiazepine drugs (BDZ) in Chile in the period 1982-1986 has been studied by the DDD method. National use was assessed using drug import forms as a source of information, assuming that the total amount imported was manufactured and consumed in the same year. The utilization did not show a constant pattern, being 32.7, 33.6, 50.2, 34.9, and 31.3 DDD 1000 per inhabitants per day in 1982, 1983, 1984, 1985 and 1986, respectively. During the study period, diazepam was the most commonly used agent amongst anxiolytic BDZ, reaching a peak of 27.1 DDD per 1000 inhabitants per day in 1984, and flunitrazepam was the most popular hypnotic, attaining its maximum in 1986 (6.4 DDD per 1000 inhabitants per day). The use of BDZ at community pharmacy level was also evaluated, employing the International Marketing System (IMS) as the source of information. At that level the pattern of utilization showed a constant increase during the study period, being 14.9 and 25.8 DDD/1000 inhabitants/per day in 1982 and 1986, respectively. In community pharmacies the anxiolytic BDZ most often consumed was diazepam (maximum 9.1 DDD per 1000 inhabitants per day in 1985), and the commonest hypnotic was flunitrazepam (peak 6.0 DDD per 1000 inhabitants per day in 1986). National consumption of BDZ appeared higher and more variable than use at the community pharmacy level, but in both there was greater usage of anxiolytic than of hypnotic BDZ, and diazepam and flunitrazepam were the most popular agents. PMID- 2571506 TI - Restricted usage of VH and V kappa genes by murine monoclonal antibodies against 3-fucosyllactosamine. AB - We are studying the structure and regulation of murine antibodies against the 3 fucosyllactosamine antigenic determinant. Analysis of the sequences of seven BALB/c IgM, kappa monoclonal antibodies (mAb), obtained from four fusions, indicates that these antibodies exhibit restriction in their usage of VH and VL genes. Based on a combination of mRNA sequences and Southern filter hybridization data, all seven light chains are encoded by V kappa 24B and J kappa 1 gene segments. Complete mRNA sequences of the heavy chains revealed that all seven mAb are encoded by VH441, six antibodies are encoded by JH4 and one uses a JH3 gene segment. The VH441 gene segment and all seven mAb contain a potential glycosylation site at Asn 58 in complementarity-determining region (CDR)2. In contrast to the similarity of the VH regions, the heavy chain CDR3 segments exhibit considerable heterogeneity. They are encoded by three D segments, they vary in length from 7-9 amino acids and display differences in their deduced amino acid sequences. The VH441 gene segment also encodes antibodies against four other carbohydrate antigens, levan, galactan, dextran and galactosyl globoside. The use of a single gene segment to encode antibodies against five different antigens suggests that the domain encoded by VH441 might be particularly well adapted for forming sites that bind carbohydrate determinants. Glycosylation of CDR2 might contribute to the unique properties of this VH domain. PMID- 2571505 TI - MALA-2, mouse homologue of human adhesion molecule ICAM-1 (CD54). AB - In humans, lymphocyte adhesion to cells is mediated by the protein heterodimer CD11a/CD18 (Leu-CAMa, LFA-1) and its ligand CD54 (ICAM-1). Although the murine CD11a/CD18 is well characterized, the mouse homologue of human ICAM-1 has not been identified. In the present study a rat monoclonal antibody to the murine lymphocyte activation antigen MALA-2 was found to inhibit in a dose-dependent manner the phorbol ester-enhanced aggregation of mouse lymphoblasts, an adhesion specific assay, and hence to define an adhesion molecule. By immunofluorescence flow cytometry the antigen expression was low on spleen cells but it largely increased after stimulation with mitogens. The antigen was expressed by some, but not all, lymphoid cell lines, and myelomonocytic and mastocytoma cells were also positive. In frozen tissue sections MALA-2 was mainly detected on germinal center B cells, dendritic cells, macrophages and vascular endothelium, including high endothelial venules. Cell surface labeling followed by immunoprecipitation and gel electrophoresis indicated that the antigen is a sialoglycoprotein which has a relative molecular mass of 95 kDa and displays a faster electrophoretic mobility under non-reducing conditions. The function, cellular distribution and molecular properties of MALA-2 are indistinguishable from those of human ICAM-1. PMID- 2571507 TI - Restriction fragment length polymorphism analysis of the V kappa locus in human lupus. AB - To evaluate the degree of genetic polymorphism of the V kappa repertoire in systemic lupus erythematosus (SLE), we performed Southern blot hybridizations with human gene probes corresponding to the four human V kappa gene families. In a comparative analysis, non-lymphoid cell DNA samples from three patients with idiopathic SLE, eight subjects with susceptibility to drug-induced lupus and seven control individuals were digested with the restriction endonucleases Bam HI, Bg 1 II, Eco RI and Hind III, and hybridized sequentially to the four V kappa family-specific probes. The restriction patterns on Southern blots revealed a low degree of polymorphism of the human V kappa gene repertoires of SLE patients and control individuals. This analysis, together with previous parallel studies of the V kappa locus in lupus-prone mice, implies that autoantibody hyperproduction in lupus is not associated with major modifications in the structure or genomic organization of immunoglobulin light chain genes. PMID- 2571508 TI - Rabbit aorta: electrical properties and agonist-induced depolarization. AB - The resting membrane potential of the smooth muscle cells of strips of rabbit aorta varied between -50 to -60 mV. No spontaneous spike discharges were observed. The membrane of the myocytes showed a marked outward rectifying property. The rabbit aorta had cable properties with a space constant (lambda) of 1.54 +/- 0.04 mm. Parallel with a progressive mechanical tension development, noradrenaline and the alpha 1-agonist methoxamine depolarized the membrane in a concentration-dependent manner up to -40 mV. Stimulation of aortic alpha 1 adrenoceptors by noradrenaline reduced the steepness of the current-voltage relationship and diminished the space constant from 1.54 to 0.8 mm, indicating a decrease in membrane resistance. No action potentials were evoked by noradrenaline. The alpha 2-agonist, B-HT 920, produced only a slight contraction and virtually no change in membrane potential. As compared to noradrenaline or methoxamine, angiotensin II was a partial agonist to induce contraction, with an intrinsic activity of 0.6-0.7. The octapeptide depolarized the membrane of the myocytes in a concentration-dependent manner and to a maximal extent similar to that observed for alpha 1-adrenoceptor stimulation. No action potentials appeared with angiotensin II. In contrast to earlier reports, depolarization did occur in the rabbit aorta in response to noradrenaline. The demonstration of depolarization raises the possibility that contraction of this blood vessel occurs through electromechanical and not or not solely through pharmacomechanical coupling when receptors for two important endogenous vasoconstrictors, noradrenaline and angiotensin II, are stimulated. PMID- 2571509 TI - Antidepressant drugs potentiate the alpha 1-adrenoceptor effect in hippocampal slices. AB - The effect of prolonged treatment with antidepressant drugs on the phenylephrine- and norepinephrine (NE)-evoked reaction in hippocampal slices was examined by extracellular recording of the spontaneous activity of CA1 layer neurons. The alpha 1-adrenoceptor agonists, phenylephrine and methoxamine, depressed the neuronal discharges of most of the units tested, while NE evoked both excitatory and inhibitory effects which were blocked by propranolol and phentolamine or prazosin, respectively. Imipramine, mianserin, (+)- and (-)-oxaprotiline administered subchronically (10 mg/kg p.o., twice daily for 14 days, withdrawal 48 h), potentiated the inhibitory reaction to phenylephrine. Mianserin was the only drug tested in the acute dose to effectively augment the reaction to alpha 1 adrenoceptor stimulation. Prolonged administration of mianserin and imipramine attenuated the excitatory effect to NE, which probably reflects beta-receptor down-regulation; however, only mianserin, but not imipramine, enhanced the NE induced inhibition. The observed potentiation of the alpha 1-adrenoceptor-related inhibitory reaction to phenylephrine produced by antidepressant drugs may reflect the development of the alpha 1-adrenergic system supersensitivity in the hippocampus. PMID- 2571510 TI - BHT-920 and LY-171555 (quinpirole) have similar affinities for striatal D-2 dopamine receptors, and similar affinities for striatal D-1 dopamine receptors. AB - The affinity of LY-171555 (quinpirole) and BHT-920 for both states of rat striatal dopamine D-1 and D-2 receptors was determined. Although these drugs have different pharmacological effects in experimental animals, we found that they had similar affinities for both D-1 and D-2 receptors. PMID- 2571511 TI - The alpha 2-adrenoceptor antagonist SK&F 104078 has high affinity for 5-HT1A and 5-HT2 receptors. AB - The affinity of SK&F 104078, a putative selective postjunctional alpha 2 adrenoceptor antagonist, was determined at 5-HT1A and 5-HT2 receptors in rat brain. SK&F 104078 had moderate affinity towards alpha 2-adrenoceptors (pKi 6.7) but displayed higher affinity at 5-HT1A (pKi 8.1) and 5-HT2 (pKi 7.6) receptors. If SK&F 104078 is used in functional studies to define heterogeneity within of alpha 2-adrenoceptors, care must be taken to define the role of 5-HT1A and 5-HT2 receptors in the response being measured. PMID- 2571512 TI - Ultrastructural immunolocalization of cyclin/PCNA in synchronized 3T3 cells. AB - The immunolocalization of cyclin/PCNA in synchronized 3T3 cells was performed with human autoantibodies using an immunogold technique performed on thin cryosections. Previous immunofluorescent studies demonstrated that the DNA replication sites correspond to the localization of bound cyclin. We have found that in the early periods of S phase, the DNA replication sites (or sites potentially ready for the replication during the hydroxyurea DNA synthesis block) are situated in the perichromatin region and correspond to clustered gold particles present frequently over a morphologically distinct small nuclear area. Heavily labeled chromocenters, including perinucleolar condensed chromatin, exhibiting several such distinct areas were found in later periods of S phase. PMID- 2571513 TI - Quantitative autoradiographic characterization of L-[3H] glutamate binding sites in rat vestibular nuclei. AB - Quantitative autoradiography has been used to characterize L-[3H] glutamate binding sites and to describe their distribution in frozen sections of rat vestibular nuclei. Scatchard plots and Hill coefficients of glutamate binding suggest that glutamate interacts with a single population of sites having a KD of about 126 nM and a capacity of 2.5 pmol/mg of protein. Although the level of glutamate binding was not very high compared to the highest levels described for some other brain regions, it was nonetheless substantial. The sites were distributed unevenly in the four vestibular nuclei and their distribution correlated well with the projection areas of the vestibular nerve, which has been described as a glutamate-mediated pathway. The highest numbers of glutamate binding sites were observed in the medial vestibular nuclei. This technique provides a very sensitive assay for characterizing the pharmacological subtypes of glutamate binding in the vestibular nuclei and for analyzing changes in these sites during development or after deafferentation of the vestibular nuclei. PMID- 2571515 TI - Xenografting of fetal pig ventral mesencephalon corrects motor asymmetry in the rat model of Parkinson's disease. AB - A suspension of cells from embryonic day 21 fetal pig ventral mesencephalon was transplanted into the striatum of 20 immunosuppressed rats with 6-hydroxydopamine induced lesions of the nigrostriatal dopamine pathway. Of these rats, 15 showed reduction of amphetamine-induced ipsilateral rotation by 9 weeks and complete reversal of rotation by 14-17 weeks. Animals maintained stable reversal of rotations (contralateral direction) until cessation of Cyclosporin A (CyA) treatment at 15-20 weeks. Within 4-9 weeks after CyA removal, these rats showed exclusively ipsilateral rotations during behavioral testing which were comparable to pre-transplant levels, suggesting that the grafts were rejected upon cessation of CyA treatment. Rats were sacrificed and tyrosine hydroxylase (TH) immunohistochemistry was performed at several time points, both on and off CyA, to examine a possible correlation between the degree of rotational behavior and the number of TH-positive surviving grafted cells. Staining showed large numbers (230-12,329) of TH-positive surviving cells in animals displaying a high degree of rotational correction (1.6 to -9.6 net ipsilateral rotations/min) after cessation of CyA treatment. Two control groups, those transplanted with non neuronal cells from the pig ventral mesencephalon (n = 5) and those receiving only daily CyA injections (n = 4) showed no significant reduction of net ipsilateral rotations throughout the experiment. No TH-positive surviving cells were seen in the one non-neuronal transplant analyzed. This data demonstrates long-term retention of xenografted tissue with immunosuppression and its concomitant restoration of normal motor behavior in the rat model of Parkinson's disease. PMID- 2571514 TI - The human locus coeruleus complex: an immunohistochemical and three dimensional reconstruction study. AB - The CA (catecholamine/catecholaminergic) cell populations of the locus coeruleus (LC) and subcoeruleus (SubC) were studied using serial sections of the human brainstem immunostained with an antibody against tyrosine hydroxylase. The tyrosine hydroxylase-immunoreactive (TH-IR) neurons were plotted in a computer reconstruction system and their number and soma size determined. Serial section computer analysis was then used to create a three dimensional reconstruction of the LC complex. The number of cells containing neuromelanin pigment was also determined and compared with the number of TH-IR cells. In our sample there were 53,900 TH-IR cells in the LC and a further 6260 cells in the SubC. These numbers were very similar to our estimates of the number of cells containing neuromelanin pigment and we concluded that virtually all of these cells were also tyrosine hydroxylase positive. The average soma size of the TH-IR cells of the LC was 37 microns and in the SubC 34 microns. In addition to these quantitative observations the morphology of the TH-IR and the Nissl stained cells is described in some detail. We also compared the groups of immunoreactive cells in the human pons with the noradrenergic groups A5-A7 described in the rat. Although in the human these groups are contiguous, A5 is not part of the LC complex. However we did find that the A7 group is equivalent to the rostroventral part of SubC while the remainder of SubC is formed by ventral A6. PMID- 2571516 TI - Effects of amygdaloid kindling on NMDA receptor function and regulation. AB - These studies were conducted to determine whether amygdaloid kindling results in the long-term alteration of NMDA receptors which could explain the persistent reduction in seizure threshold seen in this phenomenon. NMDA-induced [3H]norepinephrine (NE) release, NMDA-sensitive L-[3H]glutamate binding, and NMDA and glycine-enhanced [3H]TCP binding were measured in brain tissue from kindled rats and nonstimulated control rats 3 to 6 weeks after the last seizure. There was no difference in the ability of NMDA to induce [3H]NE release from kindled or control slices of amygdala or hippocampus. There was also no difference in the ability of phencyclidine (PCP) or Mg2+ to inhibit [3H]NE release induced by 100 microM NMDA. Equilibrium saturation experiments of NMDA-sensitive L-[3H]glutamate binding revealed no differences in KD or Bmax values between control and kindled cortex, amygdala, and hippocampus. The Ki values for NMDA displacement of L [3H]glutamate binding also did not differ in kindled tissue. NMDA-enhanced [3H]TCP binding was similar in cortex, amygdala, and hippocampus of kindled and control tissues. Finally, glycine-enhanced [3H]TCP binding was not different in control or kindled tissues. These studies suggest that the NMDA recognition site and the modulation of the NMDA receptor/ion channel complex by magnesium, PCP, and glycine are not altered several weeks after the last seizure. Even though NMDA-mediated electrophysiological responses are reportedly enhanced in kindled tissue at that time, the mechanism(s) underlying the enhancement remains to be determined. PMID- 2571517 TI - Low prevalence in Apulia (southern Italy) of HTLV I infection in drug addicts during 1986-88. PMID- 2571518 TI - [Pharmacology of various aminosteroidal muscle relaxants]. AB - A large number of analogues of pancuronium and vecuronium have been tested for neuromuscular blocking and other actions in anaesthetized cats, dogs, pigs and monkeys in an attempt to find a nondepolarizing substitute for succinylcholine. The results demonstrated an inverse relationship between dose and brevity of action; that is to say that grief duration was found only among compounds of relatively low potency. Two compounds given the code numbers Org 7617 and Org 9616 were found to possess promising properties. Both compounds are 17-butyrate analogues of vecuronium. Org 9616 also has an allyl group, in place of the methyl group, attached to the quaternary nitrogen at position 16. Both compounds have time courses similar to that of succinylcholine. They do not release histamine or produce ganglion block, and they are both considerably less potent than pancuronium in blocking the cardiac vagus. Both may produce a small fall in blood pressure. Electrophysiological analysis shows that neither compound is expected to occlude the receptor-operated ion channels of the motor endplate in concentrations likely to be achieved clinically. PMID- 2571521 TI - [The design of a feldsher center]. PMID- 2571520 TI - [Effect of carbidine on parameters of ethanol oxidation in experimental alcoholic intoxication]. AB - The effect of carbidine on enzymes of ethanol and acetaldehyde oxidation, the rate of ethanol elimination and the parameters of ethanol consumption were studied during long-term alcoholic intoxication. Carbidine administration was shown to increase the activity of alcohol dehydrogenase of the liver tissue, to decrease the activity of aldehyde dehydrogenase with a low Km to acetaldehyde. Also, the rate of ethanol elimination and a relative amount of consumed ethanol increase at the expense of an increase of the volume of consumed liquid. PMID- 2571519 TI - [Effect of various factors on the absorption of dimedrol, diprazine (pipolphen) and suprastin on rat small intestine in vitro]. AB - In the experiments on male albino rats absorption of dimedrol, pipolphen and suprastin from the small intestine was studied in vitro by the method of "turned sacks". It was shown that immunization of the animals with ovalbumin in combination with Freund's incomplete adjuvant does not influence significantly absorption of the antihistaminic agents in the rat small intestine in vitro. During anaphylactic shock absorption of pipolphen and to a lesser degree of suprastin tends to increase. The structure of the absorbing surface of the intestinal mucosal epithelium is of importance for absorption of dimedrol and to a lesser degree of pipolphen. Absorption of the antihistaminic agents in the rat small intestine in vitro depends on the activity of the drug metabolizing enzymes that especially distinctly shows up for pipolphen. Induction of the metabolizing enzymes by phenobarbital contributes to an active elimination of the agents from the incubation medium. Absorption of dimedrol and suprastin is implemented through an active transport whose energy supply for dimedrol is related to a greater extent to anaerobic processes of oxidation and for suprastin both to aerobic and anaerobic processes. PMID- 2571522 TI - [The famous pathway from feldsher to academician (Ioakim Romanovich Petrov) (1893 1970)]. PMID- 2571523 TI - [The participation of rural paramedical workers in the prevention of cardiovascular diseases]. PMID- 2571525 TI - Bio-environmental control of malaria in a tribal area of Mandla district, Madhya Pradesh, India. AB - Bizadandi block of District Mandla was selected for demonstration of bio environmental control of malaria. The project presently covers 80 experimental villages and 12 control villages. In this area, because of indifferent surveillance and spraying in the past (from 1978-86) the API was reported low and the parasite reservoir in the community had built up over the years. At the outset of the study the malaria prevalence was high with preponderance of P. falciparum. Densities of A. culicifacies were very high and A. fluviatilis was also present. P. falciparum predominated in both experimental and control area during 1988 than in 1987. Annual blood examination rate (ABER) was 73.6 and 57.5 and annual parasite incidence (API) was 114.7 and 228.0 in experimental and control villages in 1987, while in 1988, ABER and API was 63 and 73 and 112 and 316 in experimental and control, respectively. There was constant stabilization of Pf in neighbouring blocks in 1988 while in experimental areas Pf percentage has come down appreciably during March to June. It is very alarming to note that the parasite reservoir in control villages was sufficient to maintain active transmission even when antimalarial activities are at a peak. The application of residual insecticides like DDT and HCH has no tangible impact on the reduction in vector densities and the transmission of malaria. Even if a replacement insecticide like malathion is used it may produce very limited impact on vector densities in such forested zones. Thus, there is an urgent need to intensify integrated malaria control operations in the area on long term basis. PMID- 2571524 TI - [Effect of peptide-protein binding on the elimination of somatostatin and epidermal growth factor from the vascular bed by lung tissue]. AB - Somatostatin and epidermal growth factor have been studied for their elimination from the blood-circulatory channel by the lung tissue. Formation of the peptide protein complex decreases introduction of peptides into the lung tissue. Changes of the peptide-protein binding in blood plasma induce corresponding variations in transport of peptides through the hematoparenchymatous lung barrier, that can be of regulatory significance. PMID- 2571526 TI - Resting of mosquitoes in outdoor pit shelters in Kheda district, Gujarat. AB - Outdoor resting habits of mosquitoes were studied in Kheda district, Gujarat using the artificial pit shelters. Studies revealed that mosquitoes prefer to rest in pit shelters and these habitats can be used for monitoring of mosquito populations and other ecological and behavioural studies. PMID- 2571527 TI - Endotoxin inhibition of glucocorticoid enzyme induction and in vivo 3H dexamethasone labelling of rat liver nuclei. AB - 1. The effect of endotoxin on glucocorticoid (GC) induction of liver TO and TAT was investigated. 2. It was found that endotoxin inhibited not only TO GC induction, but also that of TAT, though to a lesser extent (17.41%). 3. Endotoxin did not influence the binding capacity of liver cytosol for 3H-dexamethasone at the second hour after the toxin administration. 4. In in vivo experiments endotoxin inhibited with 57.2% the binding of 3H-dexamethasone to hepatic nuclei. 5. It is suggested that the lower extent of endotoxin inhibition of GC induction of TAT may be due to the counteracting action of some inductor(s) for TAT only. PMID- 2571529 TI - Outcomes of twin pregnancies at the Hospital Central of Maputo: retrospective study of 315 consecutive twin deliveries, January 1-September 30, 1987. AB - Between January 1 and September 30, 1987, 315 (3.53% of total deliveries) consecutive twin gestations were delivered at the Department of Obstetrics and Gynaecology of the Hospital Central in Maputo Mozambique. There were 42 perinatal deaths (stillbirths + early neonatal deaths) among the 630 infants born (66.7 per 1000) and 21 early neonatal deaths among the 609 live-born infants (34.5 per 1000). Early neonatal death frequency was higher among second twins than among first ones (50 per 1000 versus 19.7 per 1000), but the difference did not reach statistical significance. Most of the second twin early neonatal deaths (11/15) were associated with a breech vaginal delivery of the first twin. PMID- 2571528 TI - Ethnic and other factors affecting birthweight in Singapore. AB - Data on 1800 term babies, 600 from each of the Chinese, Malay and Indian racial groups, were used to identify the factors affecting birthweight in Singapore. After adjustment for gestation, maternal height and other variables, the mean Indian birthweight was 100 g less than for the Chinese (P less than 0.001), 0.001), while the Malays averaged 33 g less than the Chinese. The shortfall in Indian birthweight is thought to be due, at least partly, to environmental factors. PMID- 2571530 TI - Ultrasonographic measurement of thoracic diameters of the early gestating fetus. AB - Eighty-two ultrasonographic examinations were performed on 60 of our pregnant patients with regular menstrual cycles and no complications. The pregnancies ranged from 7 to 13 weeks of gestation. Thoracic anteroposterior diameter (ETAPD), transverse diameter (ETTD), cross-sectional area (ETA) and crown-rump length (CRL) were measured on each ultrasonogram. A high correlation between CRL and gestational age was firstly confirmed in this study. ETAPD, ETTD and ETA correlated well with the gestational age and CRL, respectively. Correlations between ETAPD/CRL and ETTD/CRL ratios with the gestational age were negative. A positive correlation of ETA/CRL ratio with the gestational age was evident. Ultrasonographic measurement of the thoracic diameters of the fetus in utero should be a useful parameter to evaluate the gestational age and for early detection of growth retardation in utero. PMID- 2571531 TI - Vaginal delivery after cesarean section. AB - We retrospectively analyzed 194 pregnancies in women with a history of previous cesarean section (CS) who were offered a trial of labor. We offered every woman a trial of labor as long as she did not have a known previous classical scar. One hundred fifty-one women delivered vaginally (79%), 24 women had multiple uterine scars. Multiple gestations and breech presentation were not considered a sole indication to perform CS. Fetal and maternal morbidity are presented. We conclude that women with multiple previous CS scars can safely deliver vaginally as can women with unknown uterine scars, with careful intrapartum surveillance. Although our numbers of women with breech presentation and multiple gestations are small, in the absence of significant morbidity, we continue to allow these women to labor and deliver vaginally. PMID- 2571532 TI - Maternal mortality inquiry in a rural community of north India. AB - Community inquiry on maternal mortality was conducted in a rural area of North India. Maternal deaths were identified by multiple informants and investigated by doctors. Amongst 257 deaths registered in women in the 15-44 year age group, 55(21.4%) were maternal deaths. Maternal mortality ratio was 230 per 100,000 live births. Major causes were antepartum and postpartum hemorrhage (18.2%), puerperal sepsis (16.4%), severe anemia (16.4%), abortion (9.1%) and obstructed labor (7.3%). This rapid, simple and low cost method is recommended for application in areas where vital registration system is unsatisfactory. PMID- 2571533 TI - Antenatal screening for syphilis at the University of Nigeria Teaching Hospital, Enugu, Nigeria--a six year survey. AB - Between January 1979 and December 1984, 29,083 out of 42,515 antenatal patients booking at the Antenatal Clinic of the Department of Obstetrics and Gynaecology of the University of Nigeria Teaching Hospital, Enugu, underwent full screening for syphilis. The results showed that 890 patients (3.06%) had positive VDRL test. Of those that reacted positively in the VDRL test, 103 (11.6%) had positive TPHA test. A prevalence rate of sero-positivity of 0.35% was obtained indicating a low incidence of syphilis in our pregnant women compared with results from other parts of Nigeria. A strong recommendation was made to treat all seropositive cases whether there was historical or clinical confirmation of syphilis or not. PMID- 2571534 TI - Induction of labor by oral prostaglandin E2 in protracted pregnancy. AB - Induction of labor was performed in 20 pregnant females with postmaturity (greater than 294 days) using either oral PGE2 tablets (0.5 mg) or i.v. oxytocin drip (each group n = 10). The induction-establishment interval was significantly shorter in the oxytocin group (P less than 0.005). Moreover, the uterine activity (in Alexandria units) at 3 h post-induction and at the end of the first stage of labor, was significantly higher with i.v. oxytocin (P less than 0.005). However, the induction-delivery interval did not differ in both groups. All cases delivered spontaneously with a satisfactory Apgar score. PMID- 2571535 TI - Reproductive health of adolescents in Bangladesh. AB - The health and social effects of pregnancy in married women 13-23 years of age were studied among a population of 175,000 in rural Bangladesh. Data collected by a structured questionnaire identified pregnant women who were followed twice during pregnancy and twice postpartum. Maternal mortality for the 13-17 age group was 5.8/1000 compared to 1.8/1000 for the 18-23 year olds. Major causes of death in the younger group were obstructed labor and toxemia. The neonatal death rate was 80/1000 for the younger group and 43/1000 for the older group. The major cause of death of infants born to the younger mothers was birth injury. In infants born to older mothers, it was tetanus. Perinatal death rate was 66.4/1000. All deliveries occurred at home, and 89% of births were attended by relatives, neighbors or an untrained birth attendant. Only 13% of the women had ever used contraceptives. Use varied from 2.9% at 16 years to 24.7% at 23 years. Suggestions are made for improving maternal and child health care in rural Bangladesh. PMID- 2571536 TI - Preventive effect of traditional herbal medicine, shosaiko-to, on danazol-induced hepatic damage. AB - The incidence of the hepatic damage during treatment with danazol (D), indicated by increased serum GOT, GPT and LDH levels, has been shown to be high especially in Japan. Thus, the preventive effect of the traditional herbal medicine, shosaiko-to (SS) was investigated in the administration of D and SS for 16 weeks (D + SS group, N = 9) and the pre-administration of SS for about 4 weeks followed by D and SS for 16 weeks (SS----D + SS group, N = 15). The incidence of serum GOT, GPT and LDH levels of more than normal range during the administration of danazol in the D + SS group was similar to that in the previous study of the administration of D alone. But it was significantly lower in the SS----D + SS group than the D + SS group. The mean levels of serum GOT and GPT were much lower in the SS----D + SS group than the D + SS group throughout the administration of danazol and the difference was significant at 8, 10 and 12 weeks in serum GOT levels and at 4 weeks in serum GPT levels. These results indicate that the pre administration of SS has great efficacy in the prevention of danazol-induced hepatic damage. PMID- 2571537 TI - Successful pregnancies of two patients with relapsing thrombotic thrombocytopenic purpura. AB - Thrombotic thrombocytopenic purpura is a severe multisystemic disorder of unknown origin. The association of relapsing TTP with pregnancy is rare but well documented and high mortality rates of mothers and fetuses have been reported so far. Since the introduction of plasma therapy for treating the acute exacerbations of the disease, overall mortality rates have decreased significantly. It is now evident that the manifestations of the disease may reappear even after long disease-free intervals and as many as a third of the recovering patients may develop a relapse. Presented are two TTP patients with relapsing TTP complicating their pregnancies. Prophylactic treatment with aspirin and dipyridamole during their last three successful pregnancies prevented or minimized the severity of TTP relapses. The course of these pregnancies and the management of such patients is discussed. PMID- 2571538 TI - Hyperreactio luteinalis in normal singleton pregnancy. AB - A case of hyperreactio luteinalis in a patient with normal singleton pregnancy is reported. The course of pregnancy had been normal until the 24th week of gestation, when the mother developed lower abdominal pain and signs of virilization. She delivered of a normal female infant at 39 weeks' gestation. The baby did not show any signs of masculinization. Serum testosterone, delta 4 androstene-dione, and 5 alpha-dihydrotestosterone of the mother were markedly elevated. They remained high after the delivery but returned to the normal ranges soon after the partial resection of the enlarged ovaries. Reported causes of hyperreactio luteinalis are reviewed. Their maternal serum androgen levels were compared with cases of luteoma of pregnancy. PMID- 2571539 TI - Interstitial pregnancy: a diagnostic challenge. AB - A case report which illustrates the clinical problems and sonographic difficulties that accompany the diagnosis of interstitial pregnancy are reported. Non-specific clinical symptoms and normal sonographic findings do not always preclude interstitial pregnancy. PMID- 2571540 TI - Congenital absence of an ovary in a Nigerian woman. AB - A case of unilateral absence of the left ovary in a 45-year-old grand multiparous Nigerian woman is reported. Congenital absence of a gonad is extremely rare and it indicates a genetic or chromosomal error in the formation of the urogenital ridge. PMID- 2571541 TI - Contribution of insulin resistance to catabolic effect of prednisone on leucine metabolism in humans. AB - To determine the role insulin resistance may play in the catabolic effect of high dose prednisone therapy, healthy volunteers were studied on four occasions with the hormone-clamp technique at two insulin infusion rates. Subjects were studied after 5 days of prednisone (60 mg/day) or no steroid treatment and were infused with somatostatin, glucagon, growth hormone, [3H]glucose, [14C]leucine, and insulin (0.1 or 0.2 mU.kg-1.min-1). At each rate of insulin infusion, the rate of leucine oxidation was increased (P less than .001) after steroid treatment. Leucine flux, an indicator of whole-body proteolysis, was similar in the presence or absence of steroid treatment (2.26 +/- 0.08 vs. 2.13 +/- 0.04 mumol.kg-1.min 1, respectively) at the lower rate of insulin infusion but was higher during steroid treatment (2.18 +/- 0.06 vs. 1.84 +/- 0.13 mumol.kg-1.min-1) at the 0.2 mU.kg-1.min-1 insulin infusion. Steroid pretreatment had no significant effect on the nonoxidative rates of leucine disappearance. These data provide strong evidence that the protein wasting associated with glucocorticosteroid therapy is in part the result of steroid-induced resistance to the antiproteolytic effect of insulin and an increase in the oxidation (and thus wasting) of one essential amino acid, leucine. PMID- 2571542 TI - Role of peptide histidine isoleucine in relaxation of cat lower esophageal sphincter. AB - Vasoactive intestinal peptide (VIP) is a candidate as an inhibitory neurotransmitter mediating relaxation of the lower esophageal sphincter (LES) because VIP antiserum reduces LES relaxation in response to neural stimulation. Vasoactive intestinal peptide antiserum, however, does not completely block LES relaxation. Thus it is possible that other neurotransmitters may be involved. Peptide histidine isoleucine has structural homologies with VIP, is synthesized with VIP from a common precursor protein, coexists in some nerve cells, and is coproduced with VIP in some tumors. In numerous organ systems VIP and peptide histidine isoleucine (PHI) produce similar effects, with PHI being less potent than VIP by approximately one log number. In the LES both VIP and PHI produce tetrodotoxin-resistant dose-dependent relaxation, with PHI being almost equipotent with VIP. We therefore tested the hypothesis that PHI may be a second neurotransmitter, partly responsible for relaxation of the cat LES, by using a highly specific rabbit PHI antiserum that exhibits minimal cross-binding with VIP, secretin, and glucagon. In 3 animals, LES and brain tissue were extracted in 0.1 N HCl and assayed with a PHI radioimmunoassay. The antiserum cross-reacted with cat brain and LES showing PHI concentrations greater than 100 ng/g, with the LES containing equal or greater concentrations of PHI than brain tissue. In other animals consecutive LES circular muscle strips were cut, mounted in 1-ml muscle chambers, and stimulated with 6-s square-wave trains of 0.1-, 0.2-, 0.4-, and 0.8 ms pulses at 1, 2, and 5 Hz. These parameters produced relaxation that was completely blocked by tetrodotoxin, and reduced by VIP antiserum, but not affected by adrenergic or cholinergic receptor antagonists. Some strips were incubated in 5% or 10% PHI antiserum, whereas others were incubated in the same concentration of preimmunization serum from the same animal. Incubation in normal serum did not significantly affect relaxation, whereas in the antiserum-treated strips, LES relaxation was reduced by a significant amount (20%-30%) at all parameters of stimulation tested. Incubation in antiserum however had no effect on relaxation induced by VIP (10(-8)-10(-6) M). These data suggest that PHI may play a role in LES relaxation induced by electrical stimulation. PMID- 2571543 TI - Stress-induced changes in gastric emptying, postprandial motility, and plasma gut hormone levels in dogs. AB - The influence of acoustic stress on postprandial gastrointestinal motility, gastric emptying, and plasma gastrin, pancreatic polypeptide, motilin, and somatostatin was evaluated in conscious dogs. Six dogs were equipped with strain gauge transducers and were exposed from 1-3 h after the meal to prerecorded music (80-90 dB broad frequency noise), which produced a significant (p less than or equal to 0.05) lengthening of the gastric (31.2%) and jejunal (37.0%) postprandial pattern. In 4 other dogs with gastric cannula, a 2-h session of acoustic stress beginning just after eating a radiolabeled standard meal induced a slowing of gastric emptying of both liquid (45.7%) and solid (47.1%) phases of the test meal when measured 0.5 h after feeding. In contrast, when measured 2 h after feeding, similar values of gastric emptying of liquids and solids were observed in stressed and control animals. Compared with controls, the postprandial increases of plasma gastrin and pancreatic polypeptide levels were significantly enhanced in stressed animals and occurred early (15 min after the meal). Although postprandial decrease in plasma motilin was unchanged by acoustic stress, the rise in plasma somatostatin level was significantly (p less than or equal to 0.05) prolonged in stressed dogs. These results indicate that acoustic stress affects gastric and intestinal postprandial motility in dogs, delaying the recovery of the migrating motor complex pattern, inducing a transient slowing of gastric emptying, and enhancing the feeding-induced release of gastrin, pancreatic polypeptide, and somatostatin. Such hormonal changes might be due to a direct effect of stress rather than being the consequence of acoustic stress induced slowing of gastric emptying. PMID- 2571544 TI - Lack of an association between polymorphisms of the T-cell receptor alpha-chain and ulcerative colitis. AB - It was recently reported that, using a T-cell receptor alpha-chain complementary deoxyribonucleic acid probe (pGA5) and the restriction enzyme Eco RV, a 10 kilobase restriction fragment length polymorphism was detected significantly more frequently in patients with ulcerative colitis than in patients with Crohn's disease and controls. This finding had great potential importance, as no gene marker had previously been found to be strongly associated with inflammatory bowel disease. Therefore, an attempt to confirm it in an independent laboratory and patient population has been made in this study. Thirty patients with ulcerative colitis, 30 with Crohn's disease, and 30 healthy control subjects were studied using the Eco RV restriction enzyme and the same T-cell receptor alpha chain complementary deoxyribonucleic acid probe as was used in the prior report. No 10-kilobase fragment or any other polymorphism using this probe-enzyme combination was found in any of the individuals studied. Polymorphisms were observed with the restriction enzyme Bgl II, but their frequencies did not distinguish between cases and controls. Therefore, there is as yet no evidence for an association between polymorphisms of Eco RV-digested genomic DNA probed with the pGA5 T-cell receptor alpha-chain complementary deoxyribonucleic acid and the predisposition to inflammatory bowel disease. PMID- 2571545 TI - Migration of the myoelectric complex after interruption of the myenteric plexus: intestinal transection and regeneration of enteric nerves in the guinea pig. AB - The effects of surgical interruption of the myenteric plexus (myectomy), extrinsic denervation of a length of small intestine, or transection and reanastomosis of the intestinal wall on migration of phase III of the migrating myoelectric complex was studied in guinea pigs. In addition, the recovery of phase III migration and the regrowth of intestinal nerves and muscle across the reanastomosis was studied at various times up to 60 days after surgery. At 6-9 days after surgery, phase III did not migrate past the myectomy during 50%-60% of recorded migrating myoelectric complexes and transection and reanastomosis of the intestinal wall blocked aboral progression of phase III in 90% of cases. Extrinsic denervation did not alter phase III migration through the denervated segment. Phase III migration past the reanastomosis recovered with time after surgery; 80% recovery occurred by 60 days after surgery. Immunoreactivities for vasoactive intestinal peptide, gastrin-releasing peptide, and somatostatin were used as markers for intestinal nerves that were cut by transaction. Immunoreactivities for vasoactive intestinal peptide and gastrin-releasing peptide are contained in myenteric neurons that project in an oral to anal direction to other myenteric ganglia and to the circular muscle. Immunoreactivity for somatostatin is contained in nerve fibers projecting aborally to other myenteric ganglia. At 7-15 days after surgery, there were accumulations of immunoreactivities for vasoactive intestinal peptide, gastrin-releasing peptide, and somatostatin in nerve fibers on the oral side of the reanastomosis, but nerve fibers containing these peptides were not observed in myenteric ganglia or circular muscle close to the anal edge. At 23-28 days, immunoreactivities for vasoactive intestinal peptide, gastrin-releasing peptide, and somatostatin nerve fibers were traced across the reanastomosis and nerve terminals were detected in ganglia and muscle close to the lesion on the anal side. Nerve fibers traversed the lesion in all cases at 57-60 days and vasoactive intestinal peptide-, gastrin releasing peptide-, and somatostatin-immunoreactive nerve terminals were detected in the first two to three rows of myenteric ganglia on the anal side. Regrowth of intestinal muscle followed a similar time-course to that observed for nerves. These data suggest that interruption of the myenteric plexus alone does not completely block phase III migration. In addition, recovery of phase III migration past a reanastomosis is associated with a restoration of both nervous and mechanical connections. PMID- 2571546 TI - [The Alma-Ata Declaration and the development of sanitary and epidemiologic services in Kazakhstan]. PMID- 2571547 TI - [Functional state of the cardiovascular system as an indicator of the effect of occupational activity on white collar workers]. PMID- 2571548 TI - [3 classifications of gestosis]. AB - Three classifications: of the American College of Obstetricians and Gynecologists (ACOG), of the Organization of Gestosis (OG) and of the International Society for Study of Hypertension in Pregnancy (ISSHP) differ among one another in some essential points. According to ACOG the term "preeclampsia" means a state with hypertension and albuminuria or edema, according to ISSHP--hypertension and albuminuria. This may lead to serious misunderstanding as many obstetricians use this term to identify a directly threatening eclampsia attack. But the two classifications do not provide a term for such a condition whereas according to OG this state is identified as threatening eclampsia. The OG and ACOG classifications give a classifications according to the progression of the disease, the ISSHP does not give such a classification. ISSHP does not consider edema as a symptom, ACOG takes it into consideration if it is accompanied by hypertension according to OG the very edema not subsiding after relaxation is enough to diagnose gestosis E. ISSHP has introduced the term "hypertension in pregnancy", but also includes one-symptom form "pregnancy albuminuria" not accompanied by hypertension. Such terms suggest lack of etiological relation between those forms, which has not however been proved. The names of one-symptom forms according to OG "gestosis H" and "gestosis P" do not impose such a relation (it is a classification according to symptoms), but they do not exclude it. Contrary to ACOG and OG, ISSHP gives the level of diastolic blood pressure only. PMID- 2571549 TI - Histochemical localization of vasoactive intestinal polypeptide and its influence on contractile activity in the non-pregnant and pregnant human cervix. AB - The distribution of vasoactive intestinal peptide (VIP) was studied by immunofluorescence in cervical tissue of non-pregnant and pregnant women. VIP was localized in connection with blood vessels as well as among collagen fibres and smooth muscle cells. No difference was observed between non-pregnant and term pregnant women. The effect of VIP on cervical contractility was tested on isolated strips by superfusion in a tissue chamber. VIP inhibited contractions at 10(-8)-10(-6) M concentration, strips from term pregnant women responding more frequently at the lower concentration. It is suggested that VIP-containing neurons of the human cervix remain intact throughout pregnancy until term. PMID- 2571550 TI - [Anaphylactic shock after i.v. verapamil]. AB - A 61-year-old woman who was being treated with verapamil because of atrial fibrillation with rapid ventricular rate, went into anaphylactic shock. Verapamil is safe and effective for treating supraventricular tachyarrhythmias and serious adverse effects are uncommon. However, hypotension, cardiogenic shock, bradycardia and precipitation or aggravation of heart failure have been described. The patient presented was successfully treated with corticosteroids and antihistaminics. PMID- 2571551 TI - [Beta-blocking actions and the partial agonist activity of bopindolol, a new beta adrenoceptor antagonist, and its two metabolites]. AB - The beta-blocking effects of bopindolol, a new long-acting beta-adrenoceptor antagonist, and its two metabolites, 18-502 and 20-785, were studied in the isolated atria and trachea preparations of the guinea pig. The partial agonist activity (PAA) and the membrane stabilizing activity (MSA) were studied in the pithed rat and the right ventricular papillary muscle preparations of the guinea pig, respectively. Antagonistic effect of bopindolol against isoproterenol was 1.1-2.8 and 14.1 times more potent than propranolol in the atria and the trachea preparations, respectively, while those of 18-502 were 34.7-38.0 (in atria) and 29.0 (in trachea) times more potent than propranolol. beta-Blocking potencies of 20-785 were 50% (in atria) and 10% (in trachea) of those of propranolol. Bopindolol, 18-502 and 20-785 increased the heart rate of the pithed rat dose dependently. The effects were inhibited by pretreatment of the preparation with propranolol. The onset of action of bopindolol was slower and the duration, longer than those of its two metabolites. Bopindolol and 18-502 were more potent membrane stabilizers than propranolol, while 20-785 produced only a minimal stabilizing effect. These results indicate that not only bopindolol, but also the metabolites of bopindolol, 18-502 and 20-785 are potent beta-blocking agents with PAA. It is presumed that the long duration of action of bopindolol is due to the potent activities of its metabolites 18-502 and 20-785. PMID- 2571552 TI - [The incidence and causes of missing front teeth in children and youths]. AB - The frequency and aetiology of missing front teeth was ascertained in 1118 patients aged between ten and 20 years. Nearly every tenth patient had one ore more missing front teeth. Congenitally absent teeth accounted for 59.4% of the total. PMID- 2571553 TI - The immune response to GHRH, relationship to conformation. AB - Antibodies to GHRH1-44, GHRH1-29, and proinsulin were induced in guinea pigs. GHRH1-44 forms 7 S and 10 S complexes with antibodies. It is a divalent antigen. The sequence 30-44 bound 85% of the antibodies to GHRH1-44 with high affinity (3.8 +/- 0.9 x 10(9) l/mol). The fragment 1-29 bound with low affinity (0.6 +/- 0.3 x 10(9) l/mol) 15% of the antibodies (2p less than 0.001). Antibodies to GHRH1-29 had low affinity towards the native hormone (0.4 +/- 0.2 x 10(9) l/mol) and the region 1-29 (0.3 +/- 0.2 x 10(9) l/mol). Antibodies to proinsulin bound linear C-peptide with lower affinity (0.3 +/- 0.2 x 10(9) l/mol) than the C peptide loop in proinsulin (3.4 +/- 0.9 x 10(9) l/mol). It is concluded that the conformation of the epitopes on the sequence 1-29, recognized during the immune response, i.e. on the cell membrane, is different from the conformation of GHRH1 29 or GHRH1-44 in aqueous solution. PMID- 2571555 TI - Effect of isoproterenol on the plasma C-peptide and insulin levels in humans. AB - There are conflicting reports on the effect of stimulation of the beta-adrenergic receptors on insulin removal by the liver. It was, therefore, the aim of the present study to clarify that problem. Four experiments have been carried out on a group of 8 healthy female volunteers: (1) isoproterenol was infused intravenously, (2) glucose was infused intravenously, (3) isoproterenol was infused with glucose, and (4) infusion of isoproterenol and glucose was preceded by administration of propranolol (the beta-adrenergic blocking agent). The concentration of C-peptide and insulin was determined in plasma from the antecubital vein. It has been found that stimulation of the beta-adrenergic receptors with isoproterenol reduces insulin removal by the human liver. This effect of isoproterenol is prevented by blockade of the beta-adrenergic receptors with propranolol. PMID- 2571554 TI - Effect of biosynthetic methionyl growth hormone (GH) therapy on the immune function in GH-deficient children. AB - The ability of growth hormone (GH) to influence certain immune functions has been studied in 21 GH-deficient children aged 1.8-17.7 years, before and during therapy with biosynthetic methionyl-hGH (12 IU/m2) injected intramuscularly 3 times weekly. Blood was collected prior to GH treatment, then after 1 week, again at 3-6 months, and finally at 9-12 months of therapy. We studied (1) the distribution of the T lymphocyte subpopulations: T total (CD3), helper/inducer (CD4) and suppressor/cytotoxic (CD8) cells, using monoclonal antibodies (OKT3, OKT4, OKT8) and (2) the in vitro IgM production stimulated by pokeweed mitogen. Pretreatment CD3, CD4, CD8 values were within the normal range. They did not change after 1 week of GH therapy. Following 3-6 months of GH treatment, CD3 significantly increased (p less than 0.001), CD4 decreased (p less than 0.01), CD8 increased (p less than 0.001) and the CD4/CD8 ratio decreased (p less than 0.001). At 9-12 months of therapy, the percentages of the different groups of T cells was not significantly different from the pretreatment values. In vitro IgM production before and following 3-6 months of GH treatment was significantly lower (p less than 0.005) than that of 15 age-matched controls. At 9-12 months, GH therapy restored the in vitro IgM production. No variations in the levels of serum immunoglobulins were observed throughout the treatment period. These data suggest that GH plays a role in the development of the immune function in children. PMID- 2571556 TI - Treating a psychotic woman with changing symptoms. PMID- 2571557 TI - A systematic approach to pharmacotherapy in patients with neuroleptic-resistant psychoses. AB - The presence or absence of neuroleptic-induced pseudoparkinsonism, the presence or absence of akathisia, and plasma levels of haloperidol are used to distinguish five types of haloperidol-resistant psychotic patients. The five types are patients receiving an inadequate dose, those whose inadequate dose cannot be increased due to the presence of akathisia, those in whom neuroleptic activity at the dopamine type 2 receptors in the brain is inhibited, those receiving a bioavailable but clinically ineffective dose, and those receiving an excessive dose. Management strategies are proposed for each type of patient and include guidelines for adjusting doses of haloperidol, switching to different neuroleptics, and introducing nonneuroleptic somatic therapies. PMID- 2571558 TI - Dipeptidylaminopeptidase IV activity in T lymphocyte subsets in B cell non Hodgkin's lymphomas. AB - The total T cell population and T cell subsets in ten lymph nodes with reactive lymphoid hyperplasia (RLH) and 23 specimens (21 lymph nodes, one stomach, and one small bowel) involved by histologically and immunohistologically diagnosed B cell non-Hodgkin's lymphomas (NHLs) were determined by reactivity with monoclonal antibodies Leu 4-CD3, Leu 3-CD4, and Leu 2-CD8 in cytospin preparations from cell suspensions. T cell populations were also investigated for the coexpression of dipeptidylaminopeptidase IV (DAP IV) activity, which was visualized simultaneously with cell surface immunostaining by a combined cytochemical and immunocytochemical method. The mean absolute percentage of Leu 4-CD3+ (total T) and Leu 3-CD4+ cell populations was significantly lower in B cell NHL cases than in RLH cases (35% v 54%, P less than .001; 29.5% v 44.4%, P less than .01). No difference in the mean absolute percentage of the Leu 2-CD8+ T cell subset was found between the RLH cases and the B cell NHL cases classified as other than category A as described by the Working Formulation (WF) of NHLs. The relative percentage of Leu 4-CD3+ and Leu 2-CD8+ cells coexpressing DAP IV reactivity was lower in B cell NHL cases than in RLH cases (27.3% v 39.5%, P less than .05; 13.5% v 24.4%, P less than .10). There was no difference in the proportion of Leu 3-CD4+ cells expressing DAP IV reactivity between the NHL and RLH groups (34.5% v 36.1%). Since the mean relative percentage of Leu 2-CD8+ cells expressing DAP IV reactivity in the B cell NHL group in the other than category A according to the WF was lower than that of the RLH group (12.5% v 24.4%), and whereas the mean absolute percentage of total Leu 2-CD8+ cells was similar in the two groups (16.6% and 16.6%), a possible defective role of this Leu 2-CD8+ DAP IV+ subset, at least in B cell NHLs in the other than category A according to the WF, may be hypothesized. PMID- 2571559 TI - Different haplotypes for cystic fibrosis-linked DNA polymorphisms in Polish and Dutch populations. AB - We analyzed DNA from 34 Polish and 63 Dutch cystic fibrosis (CF) patients and their families using the polymorphic markers XV2c and KM19, which are in linkage disequilibrium with the CF mutation. Strong linkage disequilibrium was found in the Dutch population sample, but the haplotypes of the Polish chromosomes showed a significantly less extreme disequilibrium. Our data and previous studies indicate that the highest degree of homogeneity of the CF defect and hence the best possible use of the XV2c/KM19/CF linkage disequilibrium for CF carrier detection/exclusion is in populations of northern European origin. PMID- 2571560 TI - Linkage studies of the Wiskott-Aldrich syndrome: polymorphisms at TIMP and the X chromosome centromere are informative markers for genetic prediction. AB - Eleven families segregating for the X-linked recessive immune deficiency disorder, Wiskott-Aldrich syndrome (WAS), were studied by linkage analysis with an alpha satellite DNA probe, pBamX-7, which detects polymorphisms at the X chromosome centromere, locus DXZ1, as well as three other polymorphic markers defining loci on the proximal short arm of the X chromosome. Linkage has been established between WAS and DXZ1 (zeta (theta) = 7.08 at theta = 0.03) and WAS and the TIMP gene locus (zeta (theta) = 5.09 at theta = 0.0). We have also confirmed close linkage between DXZ1 and two marker loci, DXS14 and DXS7, previously shown to be linked to the WAS locus. The probe pBamX-7 detected allelic variation in all females tested, reflecting the high frequency of polymorphism at the centromere. One WAS carrier revealed a recombination between WAS and both marker loci DXZ1 and DXS14, indicating that WAS does not map between these loci. In conjunction with previous data from genetic mapping studies of WAS, these results confirm the pericentromeric Xp localization of WAS and demonstrate the usefulness of alpha satellite DNA probes as tools for genetic prediction in WAS as well as other pericentric X-linked diseases. PMID- 2571561 TI - Coincident maternal meiotic nondisjunction of chromosomes X and 21 without evidence of autosomal asynapsis. AB - A family in which the proband showed phenotypic signs of both the Turner and Down syndromes was studied cytogenetically and with restriction fragment length polymorphisms. The proband's karyotype was 46,X,+21, showing double aneuploidy without any signs of mosaicism. The single X and one chromosome 21 were of paternal origin while two chromosome 21 were of maternal origin. The nondisjunction of chromosome 21 took place in maternal meiosis II. If it is assumed that the absence of mosaicism renders postzygotic mitotic loss of the X chromosome unlikely, then the X chromosome would have been lost in maternal meiosis I or II. Recombination had occurred between the nondisjoined chromosomes 21. We conclude that double nondisjunction took place in one patient and that asynapsis was not a prerequisite for the autosomal nondisjunction. PMID- 2571562 TI - Use of variable simple sequence motifs as genetic markers: application to study of myotonic dystrophy. AB - Among the many classes of repetitive elements present in the human genome, the ubiquitous "simple sequence motifs" (SSMs) composed of [A]n, [TG]n, [AG]n or codon-tandem repeats form a major source of genetic variation. Here we report a detailed molecular-genetic study of a "variable simple sequence motif" (VSSM) in the apolipoprotein C2 (apoC2) gene, which maps to the 19q13.2 region in the vicinity of the myotonic dystrophy (DM) locus. By combining in vitro DNA amplification using the polymerase chain reaction and high-resolution gel electrophoresis, we could demonstrate a high degree of allelic variation with at least ten alleles, which differ in the number of repeated [TG] or [AG] dinucleotide units. Similar results were found for the somatostatin I gene locus. To evaluate the usefulness of SSM-length polymorphisms as genetic markers, the apoC2-VSSM was employed for linkage analysis in DM families. Our results establish that the orientation of the apolipoprotein gene cluster on 19q is cenapoE-apoC2-ter and indicate that the many thousands of structurally similar VSSMs in the human genome represent a rich source of highly informative genetic and diagnostic markers. PMID- 2571563 TI - Evidence for male X chromosomal mosaicism in X-linked agammaglobulinemia. AB - X-Linked agammaglobulinemia (XLA) is a severe antibody deficiency disease in man, resulting from an arrest in differentiation of pre-B cells. XLA is recessive: female carriers do not exhibit antibody deficiency, but manifest an exclusive inactivation of the XLA-carrying X chromosome in all peripheral blood B lymphocytes. An exclusive inactivation of the paternal X chromosome in the B lymphocytes of all daughters of a male who had no agammaglobulinemia demonstrated that the XLA defect can originate from healthy males. These males are X chromosomal mosaics. X-Chromosomal RFLP segregation analyses in other XLA pedigrees suggest a frequent introduction of XLA by healthy males. This implies that XLA often originates from mitotic errors, either at postmeiotic or early postzygotic stages. PMID- 2571564 TI - Distribution of meiotic recombination along nondisjunction chromosomes 21 in Down syndrome determined using cytogenetics and RFLP haplotyping. AB - Ten families (Down syndrome children and their parents) showing evidence of meiotic recombination between intraparental chromosomes transmitted after nondisjunction were studied. Cytogenetic polymorphisms and a cassette of RFLP markers distributed along chromosome 21 were used to analyze these families to localize the regions of meiotic recombination. Results indicated that only one crossover occurred per meiotic division and that nine of ten nondisjunctions appeared to be of maternal origin. In one family the crossover had taken place in the pericentromeric region, proximal to marker D21S13, which is quite exceptional. A chance of meiotic recombination within region 21q21, flanked by marker D21S72 and the amyloid gene, could be demonstrated in seven of the ten families. Most strikingly, this chance significantly decreased distal to q21, with frequencies of 0.3 and 0.1 in regions q22.2 and q22.3-qter, respectively. It is hypothesized that decreased chiasmata formation in the most distal part of chromosome 21q might promote nondisjunction. Furthermore, data from the ten crossovers made it possible to map provisionally two previously undefined markers, D21S24 and D21S82, to regions q21-qter and q22.1-qter, respectively. PMID- 2571565 TI - X-linked megalocornea: close linkage to DXS87 and DXS94. AB - In a family in which X-linked megalocornea is segregating, the disease locus was found to be closely linked to DXS87 (zeta max = 3.91, theta max = 0.00) and DXS94 (zeta max = 3.34, theta max = 0.00) in Xq21.3-q22. PMID- 2571566 TI - Assignment of the gene(s) involved in the expression of the proliferation-related Ki-67 antigen to human chromosome 10. AB - The antigen recognized by the monoclonal antibody Ki-67 is a proliferation related nucleolus-associated constituent used as a marker for cycling cells in tumor diagnosis. Antibody Ki-67 reacts with human proliferating cells, but not with hamster and mouse cells. Expression of the Ki-67 antigen was studied in a panel of human-rodent somatic cell hybrids. The results indicate that a gene involved in the expression of the antigen is located on chromosome 10. PMID- 2571568 TI - Physical mapping of human chromosome 17 using fragment-containing microcell hybrids. AB - Hybrid cell lines were generated by microcell-mediated transfer of human chromosome 17 into rat recipient cells. The genotypes of 36 such lines were analyzed using a set of human chromosome 17-derived sequences to probe the structural integrity of the chromosome. Four classes of hybrids were obtained: clones with an apparently intact chromosome 17, clones containing large fragments of the chromosome including both the centromere and the selected marker, clones containing only the selected marker and flanking sequences, and clones containing two 17-derived fragments--the pericentric region plus the region of the selected marker. Data from these hybrids were used in conjunction with published regional localization information to obtain a provisional linear map of the chromosome. Results of this analysis are compared to the gene maps predicted from recent linkage studies and from other somatic cell hybrid experiments. PMID- 2571567 TI - Detection of a rare allele with the pMP6d-9/MspI RFLP near the cystic fibrosis locus. AB - We report a rare allele detected using pMP6d-9, a probe very closely linked to cystic fibrosis (CF), on digestion with MspI. This allele has been found in normal and CF chromosomes, and therefore cannot be related to the mutation causing the disease. PMID- 2571569 TI - Comparison of the physical and recombination maps of the mouse X chromosome. AB - The locations of five random mouse genomic DNA markers and five cloned genes, including the genes for clotting factors VIII and IX (Cf-8 and Cf-9), Duchenne muscular dystrophy (Dmd), phosphoglycerate kinase-1 (Pgk-1), and alpha galactosidase (Ags), on the mouse X chromosome were determined by in situ hybridization. The five random DNA markers provide new genetic loci with useful restriction fragment length polymorphisms between mouse strains and species, including one locus close to the centromeric region of the mouse X chromosome. The physical map and the recombination map of these loci on the X chromosome were compared. There was good agreement in the order of loci. Relative distances between loci were consistent along the X chromosome, with the exception of the telomeric end of the long arm, where the recombination fraction observed between loci closely associated on the physical map was higher than that between similarly spaced markers located in the proximal region of the X chromosome. These results are discussed in comparison to the human X-chromosome map. PMID- 2571570 TI - Linked markers flanking the gene for multiple endocrine neoplasia type 2A. AB - The inherited cancer syndrome multiple endocrine neoplasia type 2A (MEN2A) has recently been mapped to chromosome 10. We have typed 29 families with this disorder with DNA markers from the pericentromeric region of chromosome 10. Two markers, RBP3 and MCK2, were tightly linked to the MEN2A gene at recombination fractions of less than 3%. Multipoint analysis of the linkage data suggests that the gene is located within a 3-cM interval defined by the markers RBP3/MCK2 on one side and TB14.34 on the other. No evidence for locus heterogeneity was detected in any of the 27 families from 14 countries who were informative for the markers tested. The data confirm and refine the original assignment and provide the basis for presymptomatic screening for this disorder. PMID- 2571571 TI - Localization of a human brain sodium channel gene (SCN2A) to chromosome 2. AB - A DNA probe derived from a human genomic library has been used to localize on human chromosomes a gene coding for the alpha-subunit of the brain type II sodium channel (SCN2A). Hybridization of the probe to Southern blots made with DNAs from a rodent-human somatic cell hybrid panel indicates localization to the long arm of human chromosome 2. In situ hybridization to metaphase chromosomes confirms this assignment and indicates regional localization to 2q21-q33. The probe also reveals a frequent two-allele HaeIII RFLP. PMID- 2571572 TI - A mouse linkage testing stock possessing multiple copies of the endogenous ecotropic murine leukemia virus genome. AB - A new linkage testing stock of the laboratory mouse has been constructed. The stock, designated MEV (multiple ecotropic provirus), was developed by inbreeding and selection beginning with the cross of strains C58/J and AKXD-14. Eleven different murine leukemia virus (MuLV) proviruses have been fixed in the MEV/1Ty strain. Nine of these can be uniquely identified by Southern blotting of PvuII digested DNA and probing with a cloned fragment of the ecotropic viral genome. Two proviruses had been mapped previously to chromosome 7, while single proviruses had been mapped to chromosomes 2, 9, and 11. The mapping of six additional proviruses, derived from C58/J, to chromosomes 1, 3, 5, 10, 18, and 19 is described. Another C58/J provirus was mapped to chromosome 8 and proved to be identical to the previously mapped C58v-1 virus inducibility gene of strain C58/Lw. Three dominant visible markers, hammer-toe (Hm), steel (Sl), and caracul J (CaJ), located on chromosomes 5, 10, and 15, respectively, have been introduced onto the MEV genetic background by repeated backcrosses to provide additional linkage markers. It is estimated that approximately 50% of the genome can be screened by scoring 50 fully informative gametes from a linkage cross of the MEV Hm, -Sl, -CaJ stock for the combination of viral and visible markers. A strategy for efficiently mapping new recessive visible mutations by pooling tissues for DNA extraction from mutant homozygotes among F2 progeny is described. Ways of further improving the MEV stock are discussed. The location of the Myb proto oncogene is defined relative to Sl and one of the C58/J proviruses on chromosome 10. PMID- 2571573 TI - Localization of the cryptdin locus on mouse chromosome 8. AB - Cryptdin is a defensin-related peptide, and its mRNA accumulates to high abundance in epithelial cells of intestinal crypts beginning in the second week of postnatal development. The cryptdin (Defcr) locus was assigned to mouse chromosome 8 by Southern blotting of DNAs from mouse/hamster somatic hybrid cell lines. Analysis of somatic hybrid DNAs for mouse-specific restriction fragments showed zero discordance and perfect concordance with chromosome 8. The Defcr locus was localized on chromosome 8 by analysis of DNAs from recombinant inbred (RI) strains of mice after identification of three potential Defcr alleles based on restriction fragment length polymorphisms (RFLPs) in inbred strains. The strain distribution patterns of the Defcr locus were compared with those of chromosome 8 markers in five panels of RI strains. Analysis of cosegregation of Defcr with xenotropic proviral locus Xmv-26 and additional loci confirmed the chromosomal assignment and showed that Defcr is on proximal chromosome 8 within approximately 6 (1.3 to 21.3) cM of Xmv-26. The mouse Defcr locus and the human defensin gene(s) located on chromosome 8p23 appear to map to homologous regions. PMID- 2571574 TI - Isolation, chromosomal localization, and nucleotide sequence of the human HOX 1.4 homeobox. AB - We have isolated a 14-kb DNA sequence containing a single homeobox from a low stringency screen of a human genomic phage library by using heterologous homeobox sequences as probes. Chromosomal mapping of this clone using in situ hybridization to metaphase chromosomes and a panel of mouse x human somatic cell hybrids localized it to human chromosome 7p13-p15 in the region of the HOX 1 locus. We have sequenced the homeobox and show it has 100% identity to the deduced amino acid sequence of the mouse Hox-1.4 homeobox. We detect no restriction fragment length polymorphisms with the 14-kb clone, which is devoid of any moderately repetitive DNA sequences. This implies an inability of this region to tolerate change in sequence, consistent with a function highly conserved throughout evolution. The regions in the human genome where homeobox containing loci reside share patterns of organization and sequence and have other gene loci in common, implying evolutionary constraints over these regions and providing clues on how they may have evolved. PMID- 2571575 TI - Linkage analysis of the mutation locus in the eye lens obsolescence (Elo) mouse. AB - To map precisely the mutation locus of eye lens obsolescence (Elo) on mouse chromosome 1, subsequent linkage analysis was achieved using backcross mating between 129/SvSl-Elo (Elo/+) and 129/SvSl (+/+). Mouse genomic DNAs from 17 strains including the Elo mutant mouse were first digested with several restriction enzymes and analyzed by hybridization using gamma 2- and gamma 4 crystallin cDNAs as probes. Restriction endonuclease DraI showed distinct RFLP patterns in both cases. When gamma 2-crystallin cDNA was used as the probe, two strong bands were observed at 4.0 and 2.4 kb in the majority of strains, but the former fragment shifted to the 3.4-kb position in 129/SvSl-Elo (Elo/Elo) and CFO. The polymorphism between 4.0- and 3.4-kb fragments corresponded to the gamma 1 crystallin locus (Cryg-1), and that of the 2.4-kb one, to the gamma 2-crystallin locus (Cryg-2). Mouse DNAs were also analyzed by hybridization using gamma 4 crystallin cDNA (Cryg-4). In this case, 3.4- and 3.0-kb fragments were observed in Elo and wild-type mice, respectively. The backcross offsprings were analyzed with respect to Elo, Idh-1, Cryg-1, and Cryg-4 loci. Among 223 mice analyzed, recombination between Elo and Idh-1 loci was observed in three offsprings; and that between Cryg-1 or Cryg-4 and Idh-1 loci, in one offspring. No recombination occurred between Cryg-1 and Cryg-4 alleles. PMID- 2571576 TI - Gene mapping on mouse chromosome 8 by interspecific crosses: new data on a linkage group conserved on human chromosome 16q. AB - A large conserved linkage group exists on mouse chromosome 8 and human chromosome 16q, including the loci for chymotrypsinogen B (Ctrb), haptoglobin (Hp), lecithin:cholesterol acyltransferase (Lcat), metallothionein-1,-2 (Mt-1,-2), tyrosine aminotransferase (Tat), and uvomorulin (Um). Using cloned gene probes, these six loci were mapped in M. m. domesticus X M. spretus interspecific crosses relative to a number of chromosome 8 anchor loci resulting in the gene order Es 1,Es-9-Mt-1,-2-Got-2-Es-2,Es-7,Lcat,Um-Hp,Tat,Ctrb-e. These results complement earlier studies and redefine the conserved segment on mouse chromosome 8, previously defined by the Hp-Tat interval, by the 24-cM interval between Mt-1,-2 and the conserved locus for adenine phosphoribosyltransferase, Aprt, mapped at 25 cM from Es-1 by T. B. Nesterova, P. M. Borodin, S. M. Zakian, and O. L. Serov (1987, Biochem. Genet. 25: 563-568). Within this segment, the gene order appears the same in man and mouse. While map distances between HP-TAT,HP-CTRB, and TAT CTRB of respectively 7, 11, and 9 cM have previously been measured in man, no crossovers between Hp, Tat, and Ctrb were observed in over 100 meioses in the mouse. PMID- 2571578 TI - Large-scale mapping and chromosome jumping in the q27 region of the human X chromosome. AB - We have used pulsed field gel electrophoresis for further physical mapping studies in the q27 region of the human X chromosome. We show that the DXS 102 locus and the F9 gene are separated by only 300 kb despite a genetic distance of 1.4 cM; this linkage orients our large-scale map and shows that the mcf.2 transforming sequence is telomeric to F9. A BssHII complete-digest jumping library was used to jump toward the DXS 105 locus; a 130-kb jump was achieved and the corresponding "linking clone" was obtained. PMID- 2571577 TI - A glutaminase (gis) gene maps to mouse chromosome 1, rat chromosome 9, and human chromosome 2. AB - A rat cDNA clone encoding a portion of phosphate-activated glutaminase was used to identify DNA restriction fragment length polymorphisms (RFLPs) in sets of somatic cell hybrids and between wild-derived and inbred strains of mice. Segregation of rat and mouse chromosomes among somatic cell hybrids indicated assignment to rat chromosome 9 and mouse chromosome 1. Analysis of chromosome 1 alleles for several genes in an interspecific cross between Mus spretus and C3H/HeJ-gld/gld mice indicates that glutaminase can be positioned within 5.5 +/- 2.0 cM proximal to Ctla-4. Similarly, human-hamster somatic cell hybrids were examined for RFLPs, and four human EcoRI restriction fragments were found to hybridize with the rat glutaminase probe. Two of these restriction fragments cosegregated and mapped to human chromosome 2 in a region that is syntenic with mouse chromosome 1 and rat chromosome 9. PMID- 2571579 TI - Huntington disease: no evidence for locus heterogeneity. AB - A total of 63 families with Huntington disease (HD) were examined for linkage between HD and G8 (D4S10). The families included 57 Caucasian, four Black American, and two Japanese. The combined maximum lod score was 87.69 at theta = 0.04 (99% confidence interval 0.018-0.071). The maximum frequency of recombination was 0.03 in males and 0.05 in females. Fifty-seven families gave positive lod scores; five small families gave mildly negative lod scores. The maximum likelihood estimate of alpha, the proportion of linked loci, was 1.0 with a lower 99% confidence interval of 0.88. These data suggest that there is only one HD locus, although a second rare locus cannot be ruled out. PMID- 2571580 TI - Linkage analysis in familial adenomatous polyposis: order of C11P11 (D5S71) and pi 227 (D5S37) loci at the apc gene. AB - Linkage analysis with DNA probes C11P11 and pi 227 is reported in six Scottish families with familial adenomatous polyposis. Two families were informative for C11P11 and all six were at least partly informative for pi 227. Two C11P11-apc and two pi 227-apc recombinants were identified and one of these was recombinant for both C11P11-apc and pi 227-apc. A further possible combined C11P11-apc and pi 227-apc recombination event was also identified. Peak lod score for linkage of C11P11 to apc was 5.80 at a recombination fraction (theta) of 0.069 (95% probability limits 0.012-0.191) and for linkage of pi 227 to apc was 3.19 at theta = 0.110 (95% probability limits 0.023-0.286). Peak lod score for linkage of C11P11 to pi 227 was 1.79 at theta = 0.00. The data support a gene order of pi 227-C11P11-apc. PMID- 2571581 TI - Application of natural partial digests to pulsed-field gel analysis of the amplified MDR locus. AB - The analysis by pulsed-field gel electrophoresis of partial digestion products visualized by probes for the human multidrug resistance (MDR) locus has been used to further establish the restriction map of this region. Results place the MDR1 and MDR2 genes on a single SfiI fragment, with partial digestion products establishing the distance between these genes to be 230-250 kb. The feasibility and potential advantages of using "natural" partials to generate detailed restriction maps within an amplified DNA domain are discussed. PMID- 2571582 TI - Localization of the mouse gene for secreted phosphoprotein 1 (Spp-1) (2ar, osteopontin, bone sialoprotein 1, 44-kDa bone phosphoprotein, tumor-secreted phosphoprotein) to chromosome 5, closely linked to Ric (Rickettsia resistance). AB - We have used a cDNA probe for mouse secreted phosphoprotein 1 (Spp-1, also known as 2ar, osteopontin, bone sialoprotein 1, 44-kDa bone phosphotein, tumor-secreted protein) to find a restriction fragment length polymorphism in the gene from C57BL/6J and DBA/2J mice. The strain distribution pattern in 25 BXD recombinant inbred lines is identical with that previously determined for the dominant, autosomal gene, Ricr (Rickettsia resistance). This places Spp-1 on mouse chromosome 5 with a 95% confidence limit of being within 4.32 cM of Ric. Evidence supporting the possibility of allelism between Spp-1 and Ric is briefly discussed. PMID- 2571584 TI - Unequal crossingover between homologous chromosomes is not the major mechanism involved in the generation of new alleles at VNTR loci. AB - To investigate the hypothesis that unequal exchange between homologous chromosomes is involved when new alleles are generated at VNTR loci, we used genetic linkage maps to identify flanking markers surrounding a VNTR marker locus. The minisatellite probe lambda MS1 was selected, as the hypervariable locus it detects undergoes spontaneous generation of new alleles in the germline at a rate of approximately 5%. Multipoint linkage analysis placed lambda MS1 within a cluster of polymorphic marker loci on chromosome 1p. Using the two closest flanking markers, CMM8 and YNZ2, we were able to characterize 12 new allele events in terms of crossingover between the flanking markers. Statistical analysis of these data has allowed us to reject the model that assumes that events generating new alleles always involve unequal exchange between homologous chromosomes at meiosis. PMID- 2571583 TI - The gene for retinal S-antigen (48-kDa protein) maps to the centromeric portion of mouse chromosome 1 near Idh-1. AB - S-antigen (48-kDa protein) is a soluble protein of the retina and the pineal gland that is believed to play an important role in the visual process. S-antigen is involved in the regulation of the activity of rod photoreceptor-specific cGMP phosphodiesterase (cGMP-PDE). The activity of this enzyme has been shown to be deficient in the retina of the rd mouse, which is affected by an autosomal recessive disease characterized by degeneration of the photoreceptor cells. The abnormal cGMP-PDE activity could result from, among other things, a lesion in the enzyme itself or in any of the proteins that regulate it, such as the S-antigen. We have used a mouse cDNA clone for the S-antigen to map the corresponding gene, Sag, to mouse chromosome 1 near Idh-1. Since the rd gene is located on mouse chromosome 5, our results suggest that Sag is not the site of the rd mutation. PMID- 2571585 TI - A mutant rat major histocompatibility haplotype showing a large deletion of class I sequences. AB - The LEW. 1LM1 inbred rat strain, which has been derived from a (LEW x LEW.1W) F2 hybrid, carries a major histocompatibility (RT1) haplotype which is distinct from that of the LEW strain (RT1(1)) in that certain RT1.C region-determined class I antigens are not expressed. Here we show that this phenotypic defect is due to genomic deletion of about 100 kb of the RT1.C region. Certain deleted DNA fragments have been cloned from the wild-type DNA into the EMBL4 vector. Five clones have been characterized and are shown to possess different restriction maps and to each carry a single stretch of class I cross-hybridizing sequences. Probes derived from the non-class I coding part of two clones detect fragments which are present in the wild-type but absent from the lm1 mutant. The type of deletion described here in the rat is discussed in the context of H-2D/Q deletions in the mouse. PMID- 2571586 TI - Mapping and nucleotide sequence of a new HLA class II light chain gene, DQB3. AB - A genomic clone specifying a new HLA class II antigen beta chain, DQB3, was isolated from a human genomic phage library using a DQB1 cDNA probe under low stringency conditions. Southern hybridization and nucleotide sequence analyses identified the beta 2 domain exon (exon 3) with several deleterious mutations and the CP-TM-CY exon [connecting peptide, transmembrane, and cytoplasmic regions, (exon 4)], but the first, second, and fifth exons encoding the 5' UT-leader, the beta 1 domain, and the 3' UT domain of normal beta chains, respectively, were entirely missing. The nucleotide sequences of these two exons were distinct from those of other class II beta chain genes, but slightly more related to the DQB1 and DQB2 genes than to other class II genes. The DQB3 sequence mapped between DQA2 and DQB1, 15 kb upstream from DQA2, by analysis of overlapping cosmid clones. This mapping was supported by the fact that Taq I, Msp I, and Bam HI DQB3 polymorphisms were perfectly correlated with the DQA2 polymorphism and not with any polymorphisms in the DR or DQ subregion, suggesting the presence of a hot spot for recombination between DQB3 and DQB1. PMID- 2571588 TI - DNA polymorphism of the human complement C8 beta gene: formal genetics and intragenic localization. AB - The eighth component of human complement consists of three subunits of different molecular mass, which are coded for by three separate genetic loci. Polymorphisms have been described at the protein level for the alpha and beta subunits by means of sodium dodecyl sulfate gel electrophoresis and isoelectric focusing. Using a full-length human C8 beta cDNA probe, we have studied more than 100 individuals by Southern blot analysis to detect DNA polymorphisms. We have found two restriction fragment length polymorphisms (RFLPs) with the enzymes Taq I and Bam HI. The Taq I polymorphism is defined by two alleles, i.e., a single 4.9 kb fragment or two 2.8/2.1 kb fragments. The allele frequencies are 0.68 and 0.32, respectively. The second RFLP with Bam HI is correlated with the Taq I variants: 3 kb Bam HI; 4.9 kb Taq I and 3.3 kb Bam HI; 2.8/2.1 kb Taq I. Both RFLPs could be mapped to the 3' portion of the C8 beta gene. Based on the size of genomic restriction fragments, the C8 beta gene can be estimated to have a size of 32-36 kb. Because of the even frequency distribution, the C8 beta DNA polymorphisms may be useful in gene mapping and disease association studies. PMID- 2571587 TI - A human immunoglobulin IGHG3 allele (Gmb0,b1,c3,c5,u) with an IGHG4 converted region and three hinge exons. AB - The five human IGHG genes consist of three constant domain exons plus one of or four hinge exon(s), the quadruplicated hinge region being characteristic of the IGHG3 gene. Besides this structural difference, the IGHG genes are polymorphic, as demonstrated by the restriction fragment length polymorphism and, at the protein level, by the Gm allotypic antigenic determinants. In this paper, we report the sequence of the G3m(b0, b1, c3, c5,u) IGHG3 allele, typical of the Black African populations and of populations with Negroid admixture, found in a homozygous Tunisian designated as LAT. We demonstrate that this G3 allele contains only three hinge exons instead of four (the probable result of an unequal crossing over) and that IGHG3 genes with triplicated hinge exons (and therefore encoding shorter gamma 3 chains) are present in healthy individuals from different populations. Moreover, we show that the LAT G3m (b0, b1, c3, c5, u) coding sequence results from the conversion, in the CH3 exon, of the G3m (b0, b1, b3, b4, b5, u, v) allele, the most frequent IGHG3 gene in the Negroid populations, by the homologous region of a IGHG4 gene. The structural features of the LAT IGHG3 allele, which are the lack of one hinge exon and its conversion by the IGHG4 gene, demonstrate that both crossing-over and gene conversion events occur in the evolution of the human IGHG genes. PMID- 2571589 TI - A locus telomeric to HLA-DPB encodes susceptibility to coeliac disease. PMID- 2571590 TI - Anti-ischemic effects of celiprolol in patients with exercise-induced angina pectoris. AB - The anti-ischemic properties of the new cardioselective beta-adrenoreceptor antagonist celiprolol were investigated in an open study of 12 men (mean age: 58 +/- 6.6 years) with exercise-induced angina pectoris. After all previous anti anginal medication had been withdrawn for at least 5 half-lives, the patients received placebo, single doses of nitroglycerin buccal spray, sublingual nifedipine, celiprolol alone, and then in association with nitroglycerin and nifedipine. Exercise tests were performed on a bicycle ergometer, with continuous electrocardiographic monitoring. Significantly more work was completed after treatment with celiprolol than at baseline (5280 +/- 2500 versus 4005 +/- 1792 kpm; P less than 0.01). There were further improvements in work completed after the addition of nitroglycerin and nifedipine. Celiprolol reduced the mean resting heart rate from a baseline value of 77.1 beats/min to 69.2 beats/min (P less than 0.01). In contrast, nifedipine induced tachycardia (82.6 beats/min). At rest, all treatments significantly reduced systolic blood pressure, but only nifedipine significantly reduced diastolic blood pressure. At the completion of the exercise protocol, celiprolol reduced the maximal systolic and diastolic blood pressure (P less than 0.05) with further decreases after the addition of nifedipine. The double product was significantly decreased by celiprolol compared with control, nitroglycerin and nifedipine. There was a further improvement of the double product after the addition of nitroglycerin to celiprolol, but the further increase after addition of nifedipine was not significant. In conclusion, it is clear that celiprolol, both alone and in combination with nitroglycerin or nifedipine, can significantly increase the work capacity of patients with exercise-induced angina pectoris and significantly reduce myocardial oxygen consumption. PMID- 2571591 TI - Use of progestogens in postmenopausal women. AB - Estrogen therapy for postmenopausal women has received adverse publicity since the mid-1970s because several reports linked estrogens with an increased risk of endometrial cancer. Other studies indicated that the risk of endometrial malignancy is reduced when a progestogen is added to the estrogen replacement. Not all postmenopausal women need estrogen replacement. Because some continue to produce significant amounts of endogenous estrogens, many need progestogen replacement to reduce the risk for endometrial hyperplasia and adenocarcinoma. It has been well demonstrated that estrogen replacement therapy does not increase the risk for breast cancer. However, added progestogen may actually reduce the risk for this malignancy in some women. Where estrogen therapy retards the development of and helps to prevent osteoporosis, added progestogen may restore bone which has been lost by promoting new bone formation. The greatest benefit to accrue to postmenopausal estrogen users is prevention of cardiovascular disease. Concern has been expressed that added progestogen may negate this benefit by adverse effects on lipids. Side effects of added progestogen occur, but these may be managed by changing to another progestogen or adding a mild diuretic. PMID- 2571592 TI - Safety of oral contraceptives related to steroid content: a critical review. AB - In 1980, an FDA advisory committee found inconclusive the evidence that the safety of oral contraceptives was related to their steroid content. In 1988, another FDA advisory committee examining the same evidence concluded that there is a trend to suggest such a relationship. The present review critically examines the published evidence and concludes that there is no scientific support for the suggestion that higher-dose oral contraceptives are less safe. Nevertheless, such products are no longer being made available in the United States. PMID- 2571593 TI - Unusual human chorionic gonadotropin patterns in pregnancies of undetermined location. AB - Three abnormal pregnancies demonstrating a previously unreported pattern of hCG production and clearance are reported. Further, the anatomic site of trophoblastic implantation remained unknown despite through surgical exploration. A conservative approach, including close monitoring of serum beta-hCG levels, resulted in spontaneous resolution in two cases. One women was treated successfully with methotrexate. PMID- 2571594 TI - Fertility and fetal salvage among women with uterine leiomyomas in a Nigerian Teaching Hospital. AB - Fertility and pregnancy outcome among 141 Nigerian women with uterine leiomyomas and 270 married gynaecological subjects matched for age and parity were studied. The preoperative spontaneous abortion incidence and fetal salvage among the 68 women in the study group who had previously been pregnant were 61.8% and 78.8%, respectively. Corresponding figures among women in the comparison group were 5.3% and 95.6%. Macroscopic evidence of pelvic sepsis was found at operation in 36.2% of all the women in the study group, and in 57.7% of those who complained additionally of infertility. The overall pregnancy rate following myomectomy was 37.9%, and 9.6% in those women whose presenting complaints included infertility. The mean operation-to-conception interval in the study group was 15.7 months. The fetal salvage increased to 93.5% in the women who conceived after myomectomy, and their incidence of abortion fell markedly, to 11%. Most of the deliveries (79%) in the women after myomectomy were by cesarean section. A reappearance of uterine leiomyomas occurred in 13.7% of the women. The relatively low incidence of pelvic sepsis in this series may partly be responsible for the finding of menorrhagia, rather than abdominal pain, as the commonest complaint of the women with leiomyoma. PMID- 2571595 TI - Clinical evidence of the minimal androgenic activity of norgestimate. AB - The goal in improving the progestational component of oral contraceptives (OCs) is to enhance the selectivity of the progestin by achieving a high degree of contraceptive efficacy while decreasing undesirable side effects associated with existing progestational agents. The androgenic activity of current progestins results in changes in lipid metabolism, particularly decreased levels of high density lipoprotein cholesterol (HDL), which have been associated with an increased risk of coronary heart disease (CHD). A progestin with high antiovulatory activity and minimal androgenicity would offer a clear therapeutic advantage in oral contraception. Norgestimate (NGM) is a new progestin with a unique profile of biological activity that has demonstrated a high level of selectivity in preclinical assays. The present studies were conducted to confirm clinically the low androgenic activity of NGM. Norgestimate (0.25 mg) in combination with 0.035 mg ethinyl estradiol (NGM 0.25/35) was compared with 0.30 mg norgestrel combined with 0.030 mg ethinyl estradiol (Lo/Ovral) in two multicenter clinical studies. In the first study (1,261 women), HDL levels were significantly increased from baseline levels in NGM 0.25/35 subjects but were significantly decreased in Lo/Ovral subjects. Increases in low-density lipoprotein cholesterol (LDL) levels were moderate in the NGM 0.25/35 group and pronounced in the Lo/Ovral group. A favorable lipid profile in NGM 0.25/35 subjects was also reflected in the LDL/HDL ratios, which were significantly lower in the NGM 0.25/35 subjects than in the Lo/Ovral subjects. Sex hormone binding globulin (SHBG) binds androgens, preventing clinical expression of androgenic activity. As a result, elevations in SHBG levels reduce bioactive (unbound) androgen levels and decrease the potential for androgenic side effects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571596 TI - Sex preselection in New York City: who chooses which sex and why. AB - With its increasing success, sex preselection has become a matter of general concern, and interest in it is growing. Three aspects of sex preselection in New York City were considered. Who is choosing the sex? Which sex is being chosen? Why have the choices been made? In this investigation, 178 couples were studied and all 57 non-American couples chose boys; however, 120 American couples chose boys and girls with equal frequency depending upon the gender of the children at home. Non-American couples chose boys for economic and business reasons (40%), cultural reasons (30%), and personal reasons (30%). Sex preselection is valuable in preventing sex-linked genetic disorders. Many are fearful of a sudden change in sex ratios should sex preselection be successfully and inexpensively carried out. PMID- 2571597 TI - The human sperm hypoosmotic swelling test: its practical application and suggestions for improvement. AB - Semen samples of 209 males of infertile couples were evaluated by the sperm hypoosmotic swelling (HS) test, the in vitro human sperm zona-free hamster ovum penetration assay (SPA), and by the three main parameters of the spermatogram (concentration, motility, and morphology). Whereas the HS test and SPA were poorly correlated with one another, the former showed a correlation with motility, particularly in oligo-, astheno-, and teratospermic semen samples. The simple and inexpensive HS test may prove a worthwhile tool in the diagnosis of male infertility. PMID- 2571598 TI - Reversal by human chorionic gonadotropin of the inhibitory effect of clomiphene on progesterone production by granulosa-luteal cells in culture. AB - The possibility of a direct ovarian effect of clomiphene citrate was assessed in vitro by measuring progesterone release from cultured human granulosa-luteal cells in both the presence and absence of hCG. Granulosa-luteal cells obtained by laparoscopic follicle aspiration following ovarian stimulation with hMG and hCG in women undergoing in vitro fertilization/embryo transfer were incubated for 72 hours in medium containing increasing concentrations of clomiphene (10(-9) - 10( 5) M) both without and with hCG (1 IU/mL). Clomiphene stimulated progesterone production in both the presence and absence of additional hCG at concentrations less than or equal to 10(7) M. In both the presence and absence of hCG a dose dependent inhibition of progesterone production was observed with higher concentrations of clomiphene (greater than 10(6) M). hCG (1 IU/mL) significantly increased progesterone production in control cells and at all concentrations of clomiphene (10(-9)-10(-5) M) tested. Low clomiphene (10(-9)-10(-7) M) and hCG concentrations appeared to have a synergistic effect on progesterone production. Thus it appears that clomiphene has a direct ovarian effect on progesterone production, which is stimulatory at low concentrations and inhibitory at higher concentrations. PMID- 2571599 TI - Ultrastructural analysis of the attachment sites of Escherichia coli to the human spermatozoon after in vitro migration through estrogenic cervical mucus. AB - The in vitro attachment between Escherichia coli and the human spermatozoon was studied using transmission and scanning electron microscopy, and ultracytochemistry. Samples from estrogenized cervical mucus columns containing migrating spermatozoa revealed two types of contact areas: Type I corresponded to the interaction of the bacterial fimbriae with the spermatozoal surface, and Type II, to the intermingling of the eukaryotic and prokaryotic cell glycocalyx. In both types of associations, the attachment area was lanthanum positive and displayed glycoconjugates. These types of intercellular contacts could represent the morphological basis of a mechanism by which bacteria are attached to spermatozoa and are consequently transported to the upper female genital tract. PMID- 2571600 TI - [Pagetoid reticulosis of the Ketron-Goodman type]. AB - The case of a 79-year-old woman with widespread pagetoid reticulosis (Ketron Goodman type) is presented. The nature of the epidermal cellular infiltrate in pagetoid reticulosis of the Ketron-Goodman type was examined by immunohistochemical methods and by electron microscopy. On ultrastructural examination the infiltrating cells proved to be medium-sized and large Sezary type and blastoid cells. Immunohistochemically, the epidermal infiltrate was composed of approximately 80% immature T cells (OKT 16) and 30% proliferating cells (Ki 67). Mature T cells (CD 5) were found to account for only 15-30% of all cells in the infiltrate. These results suggest that pagetoid reticulosis (Ketron Goodman type) is a malignant T-cell lymphoma, which is characterized, in addition to the epidermotropic behavior common to all T-cell lymphomas, by innidiation and in-situ spread within the epidermis. PMID- 2571601 TI - Malignant seminoma in a cryptorchid stallion. AB - A 16-year-old cryptorchid Quarter House with colic had a large, lobulated soft tissue mass to the left of the pelvic inlet. At surgery, 2 large multilobulated pedunculated masses were removed. A large blood vessel enveloped by one of the masses was damaged, and the horse exsanguinated. Postmortem examination of the abdomen revealed the masses to be malignant seminoma, with multiple sites of metastasis. PMID- 2571603 TI - The roles of CD8+ and CD4+ cells in tumor rejection. AB - In vivo administrations of anti-Lyt-2.2 (CD8) mAb and anti-L3T4 (CD4) mAb selectively eliminated CD8+ cells and CD4+ cells, respectively. The relative potencies of CD8+ cells and CD4+ cells and their roles in primary tumor rejections were studied by investigating the effects of these mAbs on tumor growth. CD8+ cells were themselves fully capable of mediating rejection in 5 different tumor rejection systems: two radiation leukemia virus (RadLV)-induced leukemias, B6RV2 and BALBRVD, a radiation-induced leukemia BALBRL male 1, and a plasmacytoma BALBMOPC-70A in CB6F1 mice, and a Friend virus-induced leukemia B6FBL-3 in B6 mice. On the other hand, CD4+ cells were capable of resisting tumor growth of B6FBL-3, but not of the other four tumors. Furthermore, for efficient rejection of CB6F1UV female 1 sarcoma by CB6F1 mice, synergy of CD8+ and CD4+ cells was necessary. Blocking of UV female 1 rejection was abrogated by delayed administration of anti-L3T4 (CD4) mAb but not anti-Lyt-2.2 (CD8) mAb, indicating the involvement of CD4+ cells in only the initial phase of rejection. PMID- 2571602 TI - Inhibition of multidrug-resistant human tumor growth in athymic mice by anti-P glycoprotein monoclonal antibodies. AB - In an effort to devise an effective treatment for human drug-resistant cancers, we have developed monoclonal antibodies, MRK16 and 17, reactive to the multidrug transporter protein, P-glycoprotein. The monoclonal antibodies given intravenously effectively prevented tumor development in athymic mice inoculated subcutaneously with drug-resistant human ovarian cancer cells 2780AD. Treatment with MRK16 induced rapid regression of established subcutaneous tumors and apparent cures of some animals. Complement-dependent cytotoxicity (MRK16) and antibody-dependent cell-mediated cytolysis (MRK16 and 17) were observed with these antibodies. These monoclonal antibodies may have potential as treatment tools against multidrug resistant human tumors possessing the P-glycoprotein. PMID- 2571604 TI - Molecular analysis of cytoplasmic genetic variation in Holstein cows. AB - Mitochondrial DNA from Holstein maternal lineages implicated to express cytoplasmic genetic effects on lactation traits was subcloned and screened for molecular polymorphisms. Sixteen of 35 lineages sampled differed from the most common mitochondrial DNA form by at least one restriction endonuclease cleavage site in the 4.3 kilobase segment examined. Variation existed in the region that regulates DNA replication and transcription as well as in transfer and ribosomal ribonucleic acid coding regions of the DNA. The index of nucleotide diversity calculated from polymorphism frequencies indicated that the minimum extent of variation between two random lineages was 1.16 x 10(-4) nucleotide differences per base pair in the segment examined. Presence of a HpaII marker near nucleotide 360 was associated with lower (P less than .001) milk fat percentages. Molecular markers indicated that pedigrees may not be sufficient to separate true cytoplasmic lineages for quantitative genetic analyses. These findings provide a molecular genetic basis for further study of cytoplasmic effects on phenotypic variation in Holstein cattle. PMID- 2571606 TI - Circulation of the airway mucosa. AB - The major part of tracheobronchial blood flow is distributed to the mucosa. Its microvasculature comprises 10-20% of the subepithelial tissue volume, with blood flow ranging from 30 to 95 ml.min-1.100 g wet tissue-1 in different animal species. Mucosal blood flow is influenced by vascular and airway pressures, inspired air conditions, and autonomic neurotransmitters. Several inflammatory mediators and neuropeptides are capable of enhancing the permeability for macromolecules in postcapillary venules and of augmenting tissue water volume, often with a concomitant increase in perfusion. These microvascular responses of the lower airway mucosa have an important role under various conditions of physiological stress and in airway inflammation. PMID- 2571605 TI - Effect of fenoldopam on preconstricted isolated salt-perfused rat lungs. AB - Using an in situ isolated salt-perfused rat lung preparation, we investigated the pulmonary vascular response to fenoldopam (a highly selective dopamine (DA1) agonist) infused at six different doses ranging from 0.1 to 10,000 micrograms/kg, during prostaglandin F2 alpha- (PGF2 alpha) induced pulmonary vasoconstriction. These experiments were repeated after selective DA1-blockade with SCH 23390. Twelve experiments were performed to evaluate the effect of fenoldopam on base line hemodynamics. Sixty experiments were performed after PGF2 alpha vasoconstriction. Thirty lung preparations were pretreated with SCH 23390. PGF2 alpha was infused into the pulmonary inflow catheter at 2.5 micrograms.kg-1.min-1 to give a sustained rise in mean pulmonary arterial pressure (5.0 +/- 1.0 mmHg). Fenoldopam, at doses of 0.1, 1, 10, 100, 1,000, or 10,000 micrograms/kg, was injected into the pulmonary artery (n = 5 blocked and n = 5 unblocked at each dose). Fenoldopam had no effect on hemodynamics in the absence of PGF2 alpha. In the unblocked group, after PGF2 alpha vasoconstriction, fenoldopam infusion resulted in a dose-dependent decrease in the mean pulmonary arterial pressure with a dose-response curve characteristic for a drug-receptor interaction [Response = -1.0 (log Dose) -1.6]. In the DA1-blocked group after PGE2 alpha vasoconstriction, the dose-response curve was shifted to the right but parallel to the unblocked group, indicating competitive receptor blockade [Response -0.8 (log Dose) -0.05]. We conclude that vasodilatory DA1-receptors are responsible for the observed results. PMID- 2571607 TI - Influence of age and exercise training on lipid metabolism in Fischer-344 rats. AB - The influence of training on fatty acid and glyceride synthesis by liver and adipose tissue homogenates of young and old Fischer-344 rats was examined. Four groups of rats (10 animals/group) were studied: young untrained, young trained, old untrained, and old trained. Training of each group was for 10 wk at 75% maximal O2 uptake. Young rats were killed at 6 mo of age and old rats were killed at 27 mo of age. Fatty acid synthesis was assessed by measuring the activities of acetyl-CoA carboxylase, fatty acid synthase, ATP citrate-lyase, "malic" enzyme, and glucose-6-phosphate dehydrogenase. Glyceride synthesis was evaluated by determining the rate of incorporation of [14C]glycerol 3-phosphate into lipids. In addition, lipoprotein lipase activity was measured in acetone-ether powders of adipose tissue from the four groups of rats. In liver, training had no effect on fatty acid or glyceride synthesis in either group. However, aging caused a significant decrease in the activities of four of the lipogenic enzymes but had no effect on glyceride synthesis. Training caused an increase in fatty acid synthase and glyceride synthesis in adipose tissue, and aging decreased lipoprotein lipase activity. It was concluded that training enhances the synthetic capacity of lipids by adipose tissue but that aging had a more profound effect in that the activities of the enzymes involved in these processes were lower in the old rats. Furthermore, the decreased activity of lipoprotein lipase in the older rats may explain the higher plasma triglyceride levels that were observed in these animals. PMID- 2571608 TI - Isolation and characterization of acetyl-coenzyme A synthetase from Methanothrix soehngenii. AB - In Methanothrix soehngenii, acetate is activated to acetyl-coenzyme A (acetyl CoA) by an acetyl-CoA synthetase. Cell extracts contained high activities of adenylate kinase and pyrophosphatase, but no activities of a pyrophosphate:AMP and pyrophosphate:ADP phosphotransferase, indicating that the activation of 1 acetate in Methanothrix requires 2 ATP. Acetyl-CoA synthetase was purified 22 fold in four steps to apparent homogeneity. The native molecular mass of the enzyme from M. soehngenii estimated by gel filtration was 148 kilodaltons (kDa). The enzyme was composed of two subunits with a molecular mass of 73 kDa in an alpha 2 oligomeric structure. The acetyl-CoA synthetase constituted up to 4% of the soluble cell protein. At the optimum pH of 8.5, the Vmax was 55 mumol of acetyl-CoA formed per min per mg of protein. Analysis of enzyme kinetic properties revealed a Km of 0.86 mM for acetate and 48 microM for coenzyme A. With varying amounts of ATP, weak sigmoidal kinetic was observed. The Hill plot gave a slope of 1.58 +/- 0.12, suggesting two interacting substrate sites for the ATP. The kinetic properties of the acetyl-CoA synthetase can explain the high affinity for acetate of Methanothrix soehngenii. PMID- 2571610 TI - Restriction fragment length polymorphism analysis of Rhizobium galegae strains. AB - Total DNA of various Rhizobium galegae strains representing different geographical origins, and taxonomic divergence was digested with three restriction enzymes separately, Southern blotted, and hybridized with six heterologous probes. The sequence divergences for different pairwise comparisons were calculated from proportions of conserved hybridizing fragments. The unweighted pair group method was used to group the strains. The symbiotic common nod and nifHDK probes used were highly conserved and grouped the strains according to the host plant, Galega orientalis or G. officinalis. The grouping derived from combined data of the constitutive hemA, glnA, ntrC, and recA probes was similar to that obtained in total DNA-DNA hybridization experiments. The constitutive probes grouped the strains in a different order than did the symbiotic probes, a result that may reflect interstrain transfer of symbiotic sequences in the course of evolution. PMID- 2571609 TI - Activation of glnA transcription by nitrogen regulator I (NRI)-phosphate in Escherichia coli: evidence for a long-range physical interaction between NRI phosphate and RNA polymerase. AB - Growth of cells of Escherichia coli in nitrogen-limited medium induces the formation of glutamine synthetase, product of the glnA gene, and of other proteins that facilitate the assimilation of nitrogen-containing compounds. Transcription from the glnAp2 promoter of the glnALG operon requires the phosphorylation of nitrogen regulator I (NRI) and, for optimal transcription, the binding of NRI-phosphate to two sites that can be over 1,000 base pairs from the binding site for RNA polymerase. In other procaryotic genes, placement of an activator-binding site further upstream from the start site of transcription diminishes expression. To determine how NRI-phosphate activates transcription and why NRI-dependent transcription differs from activation in other systems, we constructed recombinant plasmids with small alterations between the binding sites for NRI-phosphate and RNA polymerase and between the two high-affinity NRI binding sites. We demonstrate that tightly bound NRI-phosphate activated transcription from either side of the DNA helix when at least 30 base pairs separated NRI-phosphate from RNA polymerase. In contrast, activation from a partial NRI-binding site was effective only from one side of the DNA. We also observed that glnA expression was optimal when the two high-affinity NRI-binding sites were on the same side of the DNA helix. We explain these results on the basis of a hypothesis that a contact between RNA polymerase and NRI-phosphate bound to an upstream site determines the rate of glnA transcription. PMID- 2571611 TI - Cardiovascular effects and toxicity of anxiolytics and antidepressants. AB - An awareness of the cardiovascular side effect profile of anxiolytics and antidepressants is essential to the safe and effective use of these drugs. The author discusses similarities and differences among drugs and classes of drugs with regard to their effects on various aspects of the cardiovascular system, including myocardial contractility, blood pressure, and electrocardiographic changes. PMID- 2571612 TI - Clozapine for refractory schizophrenia: an open study of 14 patients treated up to 2 years. AB - Clozapine is a novel antipsychotic agent that selectively blocks mesolimbic- rather than nigrostriatal--dopamine receptors, causes fewer extrapyramidal symptoms than do other neuroleptics, and has superior antipsychotic efficacy in some patients. However, clozapine also causes agranulocytosis more frequently than do other neuroleptics. The evidence documenting the superior benefits obtained with clozapine has primarily involved short-term (4-6 weeks) trials, and the systematic evaluation of long-term clozapine use has been limited. In this study, 14 patients with refractory chronic schizophrenia were treated openly with clozapine up to 2 years; 8 did substantially better when given clozapine than they had when given other neuroleptics. That finding suggests that clozapine may provide a useful addition to the therapeutic armamentarium for the long-term treatment of schizophrenia, despite the increased risks and the need for frequent blood tests. PMID- 2571613 TI - The effect of lead on the metabolism of a nuclear matrix protein which becomes prominent in lead-induced intranuclear inclusion bodies. AB - The influence of lead and calcium on the metabolism of a nuclear matrix protein has been studied in mouse neuroblastoma 2a (Nb2a) cells. This protein, p32/6.3, has an unusual distribution in that it is relatively abundant only in normal neural tissues and in intranuclear inclusion bodies induced in kidney tubule lining cells of chronically lead-intoxicated animals (Egle, P. M., and Shelton, K. R. (1986) J. Biol. Chem. 261, 2294-2298). The mechanism(s) whereby lead increases p32/6.3 content are of interest, but its slow accumulation in intact animals over a period of weeks to months precludes studies with metabolic inhibitors. However, the enriched levels of p32/6.3 in mouse neuroblastoma 2a (Nb2a) cells permit these studies. The relative abundance of this protein was found to increase in mouse Nb2a cells after 1- and 3-day exposures to lead. This increase could be attributed to a decreased rate of degradation rather than either increased transcription or increased translation. A role for calcium in p32/6.3 regulation was also explored. Although neither increased extracellular calcium nor calcium ionophores had an effect, treatment of the cells with the calcium chelator [ethylenebis(oxyethylenenitrilo)]tetraacetic acid decreased p32/6.3 levels in a concentration-dependent manner, suggesting a role for calcium in the normal metabolism of the protein. PMID- 2571615 TI - Gold--by mouth? PMID- 2571614 TI - Trimeric binding of the 70-kD uncoating ATPase to the vertices of clathrin triskelia: a candidate intermediate in the vesicle uncoating reaction. AB - Clathrin-coated vesicles were uncoated with the 70-kD "uncoating ATPase" from bovine brain, and the molecular products were visualized by freeze-etch electron microscopy. This yielded images of released clathrin triskelia with up to three 70-kD uncoating ATPase molecules bound to their vertices. Likewise, incubation of soluble clathrin triskelia with purified uncoating ATPase also led to trimeric binding of the ATPase to the vertices of clathrin triskelia. However, this occurred only when either EDTA or nonhydrolyzable analogues of ATP were present, in which case the ATPase also appeared to self-associate. When ATP was present instead, no 70-kD ATPases could be found on clathrin triskelia and all ATPases remained monomeric. These observations support the notion that ATP controls an allosteric conversion of the 70-kD uncoating ATPase between two different molecular conformations, an ATP-charged state in which the molecule has relatively low affinity for itself as well as low affinity for clathrin, and an ATP-discharged state in which both of these affinities are high. We presume that in vivo, the latter condition is brought about by ATP hydrolysis and product release, at which point the ATPase will bind tightly to clathrin and/or self associate. We further propose that these reactions, when occurring in concert within a clathrin lattice, will tend to destabilize it by a mechanism we call "protein polymer competition". We stress the analogies between such a mechanism of uncoating and the ATP-driven events in muscle contraction. Finally, we show that under experimental conditions in which the uncoating ATPase fully removes the coats from brain coated vesicles, identical aliquots of the enzyme do not affect plasmalemmal coated pits in situ. This remarkable selectivity, the mechanism of which remains a complete mystery, is at least consistent with the idea that the 70-kD ATPase indeed plays a role in uncoating coated vesicles after they have formed in vivo. PMID- 2571616 TI - Alpha-1 blockers. A new generation of antihypertensive agents. AB - Alpha-1 blockers have certain disadvantages over conventional and antihypertensive therapies in their haemodynamic profile and metabolic effects. This paper reviews the development of alpha-blockade, the therapeutic efficacy of prazosin, the prototype alpha-1 blocker, and the rationale for the once-daily antihypertensive compounds, terazosin and doxazosin. These drugs offer a useful alternative to first- or second-line therapy in suitable hypertensive patients, particularly with their potentially beneficial effects on serum lipids. PMID- 2571617 TI - Simple and rapid method for the simultaneous determination of the eight main metabolites and conjugates of sulphasalazine in human plasma, urine and faeces using dynamically modified silica. AB - A simple and rapid method for the simultaneous determination of sulphapyridine, N acetylsulphapyridine, 5-aminosalicylic acid, N-acetyl-5-aminosalicylic acid, hydroxysulphapyridine, N-acetylhydroxysulphapyridine, sulphapyridine O glucuronide and N-acetylsulphapyridine O-glucuronide in plasma, urine and faeces is presented. After precipitation of plasma proteins by addition of methanol the samples are injected directly into the liquid chromatographic system. The limit of detection is 1 microgram/ml or less at a signal-to-noise ratio of 5 for all compounds using ultraviolet, fluorescence or electrochemical detection. The advantages of the dynamically modified silica approach in this reversed-phase high-performance liquid chromatographic method are demonstrated with respect to regulation and reproducibility of the selectivity. PMID- 2571618 TI - Rapid and simple assay method to study the content and the biosynthesis of peptide leukotrienes in whole blood. PMID- 2571620 TI - Correction of serum neuroleptic activity for blood-to-brain distribution: a method that may render radioreceptor assay results comparable between neuroleptics. AB - Serum neuroleptic activity by radioreceptor assay and prolactin concentration were measured every 6 months for 2 years in 105 male schizophrenic outpatients. The patients took a variety of neuroleptics at clinically determined doses. As expected, when four dissimilar neuroleptics were examined together, there was no significant correlation between serum neuroleptic activity and either equivalent neuroleptic dose or serum prolactin concentration. Moderate correlations were seen, however, when neuroleptic activity was standardized between neuroleptic drugs. Similar moderate correlations were seen when neuroleptic activity was corrected for the blood-to-brain distribution of each neuroleptic. These two correction techniques ranked corrected neuroleptic activity in very similar orders. Correction for blood-to-brain distribution may be a practical way to compare serum neuroleptic activity from different neuroleptics. PMID- 2571619 TI - Plasma chromogranin-A in primary hyperparathyroidism. AB - We sought an explanation for prior findings of high plasma chromogranin-A (Chr-A) in primary hyperparathyroidism. Chr-A was measured in plasma samples from 55 controls and 73 patients with primary hyperparathyroidism caused by adenoma (n = 14), sporadic or familial hyperplasia (n = 10), or familial multiple endocrine neoplasia type 1 (FMEN1; n = 49). Serum or plasma samples were also tested for calcium, PTH, gastrin, pancreatic polypeptide, CG alpha, and PRL. Plasma Chr-A was 34 +/- 10 in parathyroid adenoma, 55 +/- 33 in parathyroid hyperplasia without FMEN1, 63 +/- 88 in FMEN1, and 25 +/- 8 in controls (mean +/- SD; nanograms per ml; FMEN1 or parathyroid hyperplasia vs. control, P less than 0.05). Plasma Chr-A did not correlate with other hormonal variables in controls. Plasma Chr-A correlated with log serum gastrin (r = 0.43; P = 0.003) and plasma PTH (r = 0.52; P less than 0.05) only in FMEN1. In FMEN1, plasma Chr-A was highest in subjects with Zollinger-Ellison syndrome (ZES, 120 +/- 127; no ZES, 30 +/- 33 (P less than 0.0001). Parathyroidectomy did not decrease plasma Chr-A in patients with parathyroid adenoma or parathyroid hyperplasia. For FMEN1 patients with available pre- and postparathyroidectomy samples, Chr-A decreased postoperatively in four of five patients with ZES compared to none of six patients without ZES (P less than 0.05). Elevated plasma Chr-A is not a general feature of primary hyperparathyroidism. Elevated plasma Chr-A in primary hyperparathyroidism was restricted principally to patients who also had ZES. Primary hyperparathyroidism may influence the level of Chr-A by an effect of hypercalcemia or elevated PTH on Chr-A secretion from pancreatic islet tissue. PMID- 2571621 TI - Ascorbic acid action in neuroleptic-associated amenorrhea. PMID- 2571622 TI - Prodynorphin peptide distribution in the forebrain of the Syrian hamster and rat: a comparative study with antisera against dynorphin A, dynorphin B, and the C terminus of the prodynorphin precursor molecule. AB - The neuroanatomical distribution of the prodynorphin precursor molecule in the forebrain of the male Syrian hamster (Mesocricetus auratus) has been studied with a novel antiserum directed against the C-terminus of the leumorphin [dynorphin B (1-29)] peptide product. C-peptide staining in sections from colchicine-treated hamsters is compared to staining in sections from untreated animals. In addition, the pattern of C-peptide immunostaining in hamster brain is compared to that in the rat brain. Finally, the C-peptide immunolabeling patterns in hamsters and rats are compared to those obtained with antisera to dynorphin A (1-17) and dynorphin B (1-13). Areas of heaviest prodynorphin immunoreactivity in the hamster include the hippocampal formation, lateral septum, bed nucleus of the stria terminalis, medial preoptic area, medial and central amygdaloid nuclei, ventral pallidum, substantia nigra, and numerous hypothalamic nuclei. Although this C-peptide staining pattern is similar to dynorphin staining reported previously in the rat, several species differences are apparent. Whereas moderate dentate gyrus granule cell staining and no CA4 cell staining have been reported in the rat hippocampal formation, intense immunostaining in the dentate gyrus and CA4 cell labeling are observed in the hamster. In addition, the medial preoptic area, bed nucleus of the stria terminalis, and medial nucleus of the amygdala stain lightly for prodynorphin-containing fibers and cells in the rat, compared to heavy cell and fiber staining in the hamster in all three of these regions. In the rat there is no differential staining between tissues processed with the C peptide, dynorphin A, and dynorphin B antisera, but numerous areas of the hamster brain show striking differences. In most hamster brain areas containing prodynorphin peptides, the C-peptide antiserum immunolabels more cells and fibers than the dynorphin B antiserum, which in turn labels more cells and fibers than dynorphin A antiserum. However, exceptions to this hierarchy of staining intensity are found in the lateral hypothalamus, substantia nigra, arcuate nucleus, and habenula. The differences in staining patterns between rat and hamster are greatest when C-peptide antiserum is used; apparent species differences are present, though less pronounced, in dynorphin B- and dynorphin A immunostained material. PMID- 2571623 TI - Immunohistochemical characterization of pelvic neurons which project to the bladder, colon, or penis in rats. AB - Retrograde-tracing and immunohistochemical techniques were used in combination to investigate the types of putative transmitters in pelvic neurons that project to the bladder, colon or penis of rats. In addition, populations of axon varicosities associated with these neurons were characterized. Subpopulations of neurons in colchicine-treated major pelvic ganglia and accessory ganglia of male rats contained immunoreactivity (IR) for tyrosine hydroxylase (TH), vasoactive intestinal peptide (VIP), neuropeptide Y (NPY), or enkephalin (ENK), while types of immunoreactivity found in major groups of varicose axons were ENK, cholecystokinin (CCK), and somatostatin (SOM). Substance P (SP)-IR varicose axons were much less common. Bladder and colon neurons were similar in a number of ways. Many neurons contained NPY-IR (greater than or equal to 50%), fewer contained TH-IR (25-30%), and even fewer contained ENK-IR (5-15%) or VIP-IR (5 10%); many neurons were associated with baskets of ENK-IR varicosities (50-65%) and fewer neurons were surrounded by CCK- or SOM-IR varicosities (30-35%). Colon neurons differed from penis neurons in having a slightly larger proportion that contained ENK-IR (10-15%, compared with 1-3%). Penis neurons were markedly different from the other two groups in additional ways. More than 90% of them contained VIP-IR, whereas only 5-7% contained NPY-IR and none were immunoreactive for TH. Furthermore, although the proportion of penile neurons associated with many ENK-IR varicosities was similar to the bladder and colon neurons (45-50%), they were rarely seen close to CCK- or SOM-IR varicose axons. These studies describe similarities and differences in the histochemical properties of neurons which project to the bladder, colon, or penis and of the varicose axons associated with those neurons. This gives further insights into the possible transmitter mechanisms involved in the regulation of different pelvic functions. PMID- 2571624 TI - An immunohistochemical study of fibrous papule of the nose: 25 cases. AB - Twenty-five cases of fibrous papule of the nose were studied by light microscopy and by immunohistochemistry using a panel of 4 cell markers. These included polyclonal antibodies against S100 protein and Factor XIII-a (FXIII-a), and 2 monoclonal antibodies, MAC 387 which labels monocyte derived macrophage cells and Ulex Europaeus Agglutinin-1 (UEA-1) a pan endothelial cell marker. An increase in the number of S100 protein positive cells, particularly in the upper dermis, was observed in 3 lesions. Melanin was identified in phagocytes in the superficial dermis in 6 lesions, including those with S100 protein positive cells. In all of the papules there was a marked increase in FXIII-a labelling of dendritic connective tissue cells, including spindle, stellate and multinucleate stellate cells. Immunoreactivity with FXIII-a was especially strong in the increased mononuclear dendritic cell population (greater than 80%) seen in the mid- and upper dermis. However, only 15% of the larger multinucleate stellate cells were immunostained with FXIII-a. The results achieved with markers to the macrophage cell series (MAC 387) or endothelial cells (UEA-1) showed no significant increase in labelling of the dermal cell population compared to normal skin taken from the nose. Our study suggests that fibrous papule of the nose, a lesion of uncertain histogenesis, probably represents a proliferative reactive process consisting mainly of dermal dendritic cells as identified by FXIII-a in most of the lesions. There is some evidence that a small percentage of the dendritic cells may represent involuted naevi and be of melanocytic origin. PMID- 2571625 TI - Brief psychotherapy for posttraumatic stress disorders. AB - A large-scale study of the effectiveness of psychotherapeutic methods for the treatment of posttraumatic stress disorders was conducted. The sample consisted of 112 persons suffering from serious disorders resulting from traumatic events (bereavement, acts of violence, and traffic accidents) that had taken place not more than 5 years before. Trauma desensitization, hypnotherapy, and psychodynamic therapy were tested for their effectiveness in comparison with a waiting-list control group. The results indicated that treated cases were significantly lower in trauma-related symptoms than the control group. PMID- 2571626 TI - Clinical observations of mosquito bite reactions in man: a survey of the relationship between age and bite reaction. AB - To evaluate the mechanism of mosquito bite reaction in man, the reaction to Aedes albopictus was observed in 162 subjects ranging in age between 1 to 68 years old. Bite reactions were found to consist of both an immediate and a delayed reaction. The eruption and time course of the immediate reaction were consistent with type I hypersensitivity. The eruption and time course of the delayed reaction were consistent with cutaneous basophil hypersensitivity. Positive rates of immediate reaction increased from early childhood to adolescence and decreased with age from adulthood. The appearance and intensity of the delayed reaction decreased with age. Mosquito bite reactions in human beings exposed continuously and regularly are known to change from stage 1 to stage 5 (stage 1; no reaction, 2; delayed reaction only, 3; immediate and delayed reaction, 4; immediate reaction only, 5; no reaction). Analysis of the relationship between age and bite reaction in this study indicated that the principle held true even when the exposures were irregular or at random. PMID- 2571628 TI - Modulation of in vitro synthesis of immunoglobulin and the induction of suppressor activity by therapy with intravenous immune globulin. AB - Intravenous immune globulin has become an important modality of replacement therapy for children and adults with hypogammaglobulinemia or agammaglobulinemia. It is also evident that intravenous immune globulin is effective in patients with a variety of automimmune disorders. The present study was undertaken to evaluate the effects of intravenous immune globulin replacement therapy in patients with common variable hypogammaglobulinemia. We measured pokeweed mitogen-induced immunoglobulin synthesis of patients' peripheral blood mononuclear cells cocultured in equal numbers with normal subjects' lymphocytes to assess suppressor cell activity. The mean suppressor cell activity in coculture experiments at baseline of nine patients in the study was -16%. Suppressor activity was demonstrated in two of nine patients at baseline who received intravenous immune globulin therapy before initiation of the study. After intravenous immune globulin therapy at a low dose (100 to 200 mg/kg/mo), suppressor cell activity was demonstrated in all nine patients, which ranged from -63% to -100%. In five patients intravenous immune globulin therapy was discontinued for 4 months. There was a return to baseline levels, with suppressor activity ranging from -20% to -50%. On reinstitution of high-dose intravenous immune globulin therapy (300 to 400 mg/kg/mo), all patients except one again demonstrated increased suppressor cell activity. Cell separation and recombination cell culture experiments were performed. All patients had an intrinsic B cell defect before the initiation of therapy so that the effects of intravenous immune globulin therapy on B cell function could not be assessed. Combining normal B cells with the patient's T cell-enriched fraction showed marked suppression of pokeweed mitogen-induced immune globulin synthesis. This suppression was reversible by irradiation of the T cell fraction. Removal of CD8 positive T cells by cell sorting resulted in normal immunoglobulin synthesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571627 TI - Cetirizine: a pharmacokinetic and pharmacodynamic evaluation in children with seasonal allergic rhinitis. AB - In a double-blind, randomized, parallel-group 5-week study, cetirizine, 5 mg or 10 mg daily, was ingested by 10 and nine children, respectively. Cetirizine was rapidly absorbed with mean peak cetirizine concentrations of 427.6 +/- SD, 144.2 ng/ml, 1.4 +/- 1.1 hours after the 5 mg dose, and 978.4 +/- 340.6 ng/ml, 0.8 +/- 0.4 hours after the 10 mg dose. The dose-independent serum-elimination half-life of cetirizine was 7.1 +/- 1.6 hours after cetirizine, 5 mg, and 6.9 +/- 1.6 hours after cetirizine, 10 mg. Urinary excretion of unchanged cetirizine during 24 hours after the initial dose of cetirizine, 5 mg, was 40 +/- 15%, and after cetirizine, 10 mg, it was 39 +/- 14%. The mean histamine-induced wheal-and-flare areas were significantly suppressed from 1 to 24 hours after the first dose of cetirizine, 5 mg, and from 1/2 to 24 hours after the first dose of cetirizine, 10 mg, compared to the mean predose wheal-and-flare areas (p less than 0.01). During daily dosing with cetirizine, 5 mg or 10 mg at bedtime for 35 days, serum cetirizine concentrations and suppression of histamine-induced wheals and flares were monitored every 7 days, 12 hours after the cetirizine dose. The mean serum cetirizine concentrations remained relatively stable during this time, and the mean wheal-and-flare areas remained significantly suppressed (p less than 0.01) compared to baseline wheal-and-flare areas measured before the first dose of cetirizine. The symptoms and signs of allergic rhinitis were suppressed throughout the study by cetirizine, 5 mg and 10 mg.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571629 TI - Clinical significance of anti-TSH antibody in sera from patients with Graves' disease and other thyroid disorders. AB - In a survey of patients having anti-TSH antibody (TSH Ab), data from 167 subjects were collected from 8 Japanese Institutions. They were divided into a high TSH Ab group consisting of 63 cases; since the means of assay was via a subnormal thyrotropin binding inhibitor immunoglobulin (TBII) assay, this group had TBII values less than -20%. An additional low TSH Ab group was made up of 104 cases. Out of a total of 11,211 patients, the incidence of TSH Ab for the high and low groups were 0.57% and 13.4%, respectively. More than 95% of these TSH Ab carriers had Graves' disease or some other autoimmune thyroid disorder, and anti thyroglobulin and anti-thyroid microsomal antibodies were detected similarly in both groups. It was significant that TSH receptor antibodies could also be detected in both groups, namely, thyroid stimulating antibody and long acting thyroid stimulator (LATS) in 4 of 9 patients in the high TSH Ab group and TBII in 55 of 104 in the low TSH Ab group, respectively. The high TSH Ab levels tended to persist, but 26% of cases showed disappearance or appearance of the antibody during the observation period. In one Graves' patient, a moderate TBII activity (64.2%) was followed by markedly elevated TSH Ab (TBII: -83.4%) within 2 months. The TSH Ab in the low TSH Ab group disappeared in most cases. Also, fluctuations in TSH Ab did not always parallel those seen for TBII and reciprocal fluctuation pattern (transient or otherwise) were observed in 33%. In conclusion, anti-TSH antibody is produced frequently in patients with either Graves' disease or some other autoimmune thyroid disorder.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571631 TI - Dystrophin-related muscular dystrophies. AB - The gene for the locus involved in Duchenne and Becker muscular dystrophies has been cloned and subject to intense analysis. The protein product of the locus is called dystrophin, and it has been shown to be associated with the muscle fiber membrane. The new knowledge of the molecular genetics of these disorders is being applied rapidly in clinical practice. Carrier detection and prenatal diagnosis have been revolutionized by the use of probes for the gene. These probes are also being employed to clarify cases where conventional clinical examination results in equivocal diagnoses. It is suggested that the disorders characterized by dystrophin abnormalities should be called dystrophin-related muscular dystrophies (DRMD). There are mouse and dog models for DRMD and these are being used to explore therapeutic strategies for treating DRMD patients. PMID- 2571630 TI - Paracrine and autocrine functions of glomerular mesangial cells. PMID- 2571633 TI - Gallbladder vasculitis. AB - Five cases of gallbladder vasculitis were encountered during a 10-year period in a community hospital with annual surgical cases of about 12,000. All five patients were clinically diagnosed as having cholelithiasis with or suspected to have cholecystitis. Vasculitis in the form of arteritis identical to that seen in polyarteritis nodosa was present microscopically in the cholecystectomy specimens. One patient presented with systemic symptoms, and the diagnosis of polyarteritis nodosa was established after arteritic lesions were identified in the cholecystectomy, liver biopsy, and appendectomy specimens. In another patient, the diagnosis of polyarteritis nodosa was established after a retrospective identification of one additional site of arteritis in the appendix removed 5 years prior to cholecystectomy. In another, gallbladder vasculitis was associated with scleroderma; the patient was studied thoroughly at autopsy shortly after cholecystectomy, and no evidence of polyarteritis nodosa was found. In the last two, gallbladder vasculitis was not associated with systemic diseases. PMID- 2571632 TI - Neurologic sequelae of pertussis immunization--1989. PMID- 2571634 TI - Prolonged use of neuroleptics in schizophrenia: a review for the practitioner. PMID- 2571635 TI - Treatment of resistant depression. Review on the efficacy of various biological treatments, specifically in major depression resistant to cyclic antidepressants. AB - The biological treatment of depression includes administration of psychoactive drugs (cyclic antidepressants, MAO-inhibitors, neuroleptics and lithium), use of certain substances which in small amounts are normally present in food such as, L tryptophan (L-TP) and L-5-hydroxytryptophan (L-5HTP), electroconvulsive therapy (ECT) and various manipulations of the sleep-wake rhythms. This paper reviews the literature on the efficacy of these treatments in patients resistant to earlier adequate treatment(s) with cyclic antidepressants. Subsequently the following strategy for the biological treatment of (non-psychotic) major depression is suggested: (1) administration of a cyclic antidepressant; (2) if after a period of 4 to 6 weeks a patient has not responded to an adequate dose, another cyclic antidepressant should be tried, adding lithium if the patient still does not respond; (3) MAO-inhibitors and (4) ECT. In psychotic depression the suggestions for the first, third and fourth steps are the same. In the second step, the cyclic antidepressant should be combined with a neuroleptic. PMID- 2571636 TI - Somatic cell mapping of the bovine somatostatin gene. PMID- 2571637 TI - Maturational steps of bone marrow-derived dendritic murine epidermal cells. Phenotypic and functional studies on Langerhans cells and Thy-1+ dendritic epidermal cells in the perinatal period. AB - The adult murine epidermis harbors two separate CD45+ bone marrow (BM)-derived dendritic cell systems, i.e., Ia+, ADPase+, Thy-1-, CD3- Langerhans cells (LC) and Ia-, ADPase-, Thy-1+, CD3+ dendritic epidermal T cells (DETC). To clarify whether the maturation of these cells from their ill-defined precursors is already accomplished before their entry into the epidermis or, alternatively, whether a specific epidermal milieu is required for the expression of their antigenic determinants, we studied the ontogeny of CD45+ epidermal cells (EC). In the fetal life, there exists a considerable number of CD45+, Ia-, ADPase+ dendritic epidermal cells. When cultured, these cells become Ia+ and, in parallel, acquire the potential of stimulating allogeneic T cell proliferation. These results imply that CD45+, Ia-, ADPase+ fetal dendritic epidermal cells are immature LC precursors and suggest that the epidermis plays a decisive role in LC maturation. The day 17 fetal epidermis also contains a small population of CD45+, Thy-1+, ADPase-, CD3- round cells. Over the course of 2 to 3 wk, they are slowly replaced by an ever increasing number of round and, finally, dendritic CD45+, Thy 1+, CD3+ EC. Thus, CD45+, Thy-1+, ADPase-, CD3- fetal EC may either be DETC precursors or, alternatively, may represent a distinctive cell system of unknown maturation potential. According to this latter theory, these cells would be eventually outnumbered by newly immigrating CD45+, Thy-1+, CD3+ T cells--the actual DETC. PMID- 2571638 TI - IL-6 and IL-1 synergize to stimulate IL-2 production and proliferation of peripheral T cells. AB - Purified T cells can be induced to proliferate and to produce the autocrine growth factor IL-2 with mAb to the TCR and costimulatory cytokines. In a previous report we demonstrated that human IL-6 stimulates IL-2 production and proliferation of purified T cells, in conjunction with the insolubilized anti-TCR V beta 8 mAb, F23.1. Here we show that when CD4+ T cells are rigorously purified to greater than 99% CD4+CD8-, they respond only weakly to F23.1 and IL-6. Instead, there is an additional requirement for IL-1, which dramatically synergizes with IL-6 to induce prolonged (greater than 7 days) proliferative responses and IL-2 production. Similar results were observed when the highly mitogenic anti-CD3 mAb 145-2C11 was substituted for F23.1. The proliferation induced by F23.1, IL-1, and IL-6 was substantially (greater than 80%) inhibited by a mAb to mouse IL-2, and was not inhibited by an anti-IL-4-mAb. In accordance with this finding, medium conditioned by the activated CD4+ cells contained large amounts of IL-2, which increased over a 7-day culture period. These results demonstrate that IL-6 and IL-1 stimulate T cell proliferation by inducing production of the autocrine growth factor IL-2. In addition, the two lymphokines must be present simultaneously for activation to occur. The possible roles of IL 6 and IL-1 in IL-2 gene regulation and in Ag-induced T cell activation are discussed. PMID- 2571640 TI - [Surgery of the adrenal glands in Sipple's syndrome. Apropos of 8 cases]. AB - In a series of 82 pheochromocytomas operated on, the authors report 8 cases of Sipple's syndrome, currently classified by Steiner among the type II multiple endocrine neoplasia (MEN II). Six patients were operated on, the mean follow up being 4 years. These patients all underwent total thyroidectomy and 4 of them total bilateral adrenalectomy. Gluco and mineralocorticoid and thyroid hormone replacement therapy was instituted. No clinical or biological recurrence has been noted. The ideal therapeutic schema is difficult to define given the rarity of MEN II. PMID- 2571639 TI - Incorporation of bacterial peptidoglycan constituents into macrophage lipids during phagocytosis. AB - It has previously been established that several glycopeptides of peptidoglycan origin are formed as a result of processing of Bacillus subtilis cell walls by the macrophage-like cell line RAW264. Although the formation of these glycopeptides could account for the humoral immune responses characteristic of bacterial peptidoglycans, their formation does not account for the cellular mediated immune responses observed for water-in-oil emulsions of peptidoglycan or for lipophilic derivatives of glycopeptide fragments thereof. Therefore, the processing of peptidoglycan by macrophages was reexamined to establish whether the lipophilic derivative of any peptidoglycan-derived glycopeptide was formed. The experiments were performed by incubating B. subtilis cell walls radiolabeled in muramic acid, glucosamine, alanine, glutamic acid, and diaminopimelic acid residues in the presence of the macrophage-like cell line RAW264. The crude lipid fraction derived from the macrophages was further fractionated and analyzed, revealing the presence of two lipophilic glycopeptides that contained glucosamine, muramic acid, and alanine of bacterial origin. PMID- 2571641 TI - Melanogenic regulatory factors in coated vesicles from melanoma cells. AB - Coated vesicles have been found to contain much higher tyrosinase and gamma glutamyl transpeptidase activities than premelanosomes. This indicates that similar to tyrosinase, gamma-glutamyl transpeptidase, an enzyme responsible for pheomelanogenesis, is highly concentrated in coated vesicles after its maturation in Golgi associated endoplasmic reticulum (GERL). Furthermore, in the pre- and post-dopaquinone melanogenic pathway, coated vesicles convert dopachrome to colorless indole compounds more quickly than in premelanosomes because of their higher dopachrome conversion factor activity. Melanosomes have been found to exhibit indole conversion factor activity, while coated vesicles show indole blocking factor activity. In moderately tyrosinase-rich premelanosomes, the levels of dopachrome conversion factor and indole blocking factor are lower than in coated vesicles or melanosomes. High levels of indole blocking factor in coated vesicles may indicate why melanin polymer formation does not occur there in vivo despite their high tyrosinase activity. PMID- 2571642 TI - Characterization of primary T helper cell activation and T helper cell lines stimulated by hapten-modified, cultured Langerhans cells. AB - It has recently been shown that hapten-modified cultured Langerhans cells are able to activate small resting syngeneic L3T4+ T helper cells from nonsensitized animals. Repeated stimulation of these T cells with hapten-modified cultured Langerhans cells leads to the establishment of L3T4+ hapten-specific interleukin 4-producing T-cell lines. Here we report on further characteristics of primary hapten-dependent activation of L3T4+ T cells and of T-cell lines derived from them. Dendritic cell-enriched spleen cells were as able as Langerhans cells to activate nonsensitized T helper cells after hapten modification. However, M12c, a major histocompatibility complex class II-positive B-cell line that was able to activate small, resting, allogeneic L3T4+ T cells was not able to stimulate syngeneic T helper cells after hapten modification. Thy1+ dendritic epidermal cells did not significantly affect the magnitude of primary T helper cell proliferation induced by cultured Langerhans cells. Restimulation of in vitro primed T helper cells with hapten-modified cultured Langerhans cells revealed the presence, within the primed T helper cell population, of activated cells with specificity to an unrelated hapten, suggesting that, in hapten-dependent T helper cell activation, hapten-nonspecific cells are activated along with those that are hapten specific. Restimulation of a hapten-specific long-term T helper cell subline using different antigen-presenting cell types demonstrates that factors other than major histocompatibility complex class II density or tissue derivation of the antigen-presenting cell play a role in the activation of T cells in vitro. Finally, we demonstrate that in vitro generated hapten-specific T helper cell lines may not show strict major histocompatibility complex restriction. PMID- 2571644 TI - Expression of proliferating cell nuclear antigen/cyclin in human keratinocytes. AB - The expression of proliferating cell nuclear antigen (PCNA), also called cyclin, in human keratinocytes was examined by using the serum obtained from a SLE patient and a murine monoclonal antibody against PCNA/cyclin. In the normal epidermis, few of the nuclei were labeled with anti-PCNA/cyclin. This was in contrast to the positive nuclear staining seen in active lesions of psoriasis. In a primary culture of human keratinocytes growing as a monolayer, 20%-30% of cells expressed PCNA/cyclin. SV40-transformed human keratinocytes showed positive nuclear staining in about 40% of the cell population. In stratified keratinocytes cultured in a high Ca++ medium, PCNA/cyclin expression was decreased and only the cells in the basal and suprabasal layers showed positive staining. These results indicate that the expression of PCNA/cyclin correlates with the proliferating state in human keratinocytes and may not be associated with the mechanism of differentiation in keratinocytes. PMID- 2571643 TI - Intercellular adhesion molecule expression in the evolving human cutaneous delayed hypersensitivity reaction. AB - Intercellular adhesion molecule-1 (ICAM-1), putatively expressed by antigen presenting or target skin cells, is a ligand for the lymphocyte function associated antigen (LFA-1) present on circulating lymphocytes. Immunohistochemistry of normal adult human skin using monoclonal antiserum to ICAM-1 demonstrated focal reactivity restricted to endothelium lining the dermal microvasculature. Delayed hypersensitivity responses elicited with dinitrochlorobenzene in the skin of the same subject were evaluated sequentially over a 96 h period using immunohistochemical and ultrastructural techniques. The first alteration observed consisted of mast cell degranulation within perivenular foci in the superficial dermis at 4 h after antigen challenge. Sparse superficial perivascular T-cell infiltrates were present by 24 h. Progressive staining for ICAM-1 was observed in microvascular endothelium and in dermal dendritic cells between 24 and 48 h. ICAM-1 expression was documented focally within the lower epidermis at 48 h and diffusely within the lower and upper epidermal layers at 96 h. ICAM-1 expression by keratinocytes was consistently associated with T-cell migration into the epidermis, whereas migration was never observed in the absence of ICAM-1 reactivity. Immunoelectron microscopy confirmed ICAM-1 to be exclusively present on endothelial cells, dermal dendritic cells, mononuclear cells, and keratinocytes, and permitted characterization of the patterns of membrane reactivity. ICAM-1 expression by epidermal cells appears to be closely linked to the progressive migration of T cells from the dermis into the epidermis that characterizes cutaneous delayed hypersensitivity. PMID- 2571645 TI - T-cell clones with L3T4-positive or Lyt-2-positive phenotypes responding to mutant MHC class II antigen and inducing graft versus host reaction. AB - Two types of T cell clones responding to mutant major histocompatibility class II antigen (Iabm12) were established from spleen cells of C57BL/6 mice: one was L3T4 positive and the other Lyt-2-positive. These two types of clones carried functionally different properties. Lyt-2+ clones were absolutely dependent on exogenous interleukin-2 for their proliferation, whereas some L3T4+ clones secreted interleukin-2 and proliferated autonomously. Both types of clones had cytotoxic activities to bm12 target cells, and Lyt-2+ clones showed stronger activities than L3T4+ clones. Lyt-2+ clones induced induration in situ, whereas the L3T4+ clones induced ulcerative reaction when injected intradermally into mice. Histologically, the L3T4+ clones caused necrosis of the epidermis or upperdermis, while the Lyt-2+ clones induced infiltration of small round cells through the epidermis to the subcutaneous tissues and caused thickening of the epidermis. These characteristic reactivities might be due to a difference in lymphokines produced by each type of T cell subset in response to Iabm12 antigen. PMID- 2571646 TI - The effect of concurrent human immunodeficiency virus infection on chronic hepatitis B: a study of 150 homosexual men. AB - To determine the influence of concurrent human immunodeficiency virus (HIV) infection on chronic hepatitis B virus (HBV) infection, 150 male homosexual chronic hepatitis B surface antigen (HBsAg) carriers were studied. Of these, 82 subjects (55%) tested positive for antibodies to HIV. They were more likely to express hepatitis B "e" antigen (HBeAg) (P less than .001) and HBV-DNA (P less than .0005) in serum than were HIV-seronegative individuals. However, the degree of immune suppression did not influence HBeAg-HBV-DNA expression. In HBeAg seropositive subjects, concurrent HIV infection was associated with lower serum alanine transferase levels (P less than .001). This effect increased with the degree of immune suppression as determined by CD4+ lymphocyte counts. Conversely, in patients negative for HBeAg, there was a weak trend towards higher alanine transferase levels with concurrent HIV. This study suggests that chronic hepatitis B may be less severe when accompanied by HIV infection; however, greater viral replication may make it more contagious and resistant to antiviral therapy. These data support an immune-mediated pathogenesis for hepatitis B and have implications for its control. PMID- 2571647 TI - Pseudomonas osteochondritis complicating puncture wounds of the foot in children: a 10-year evaluation. AB - From 1978 to 1988, microbiologically proven Pseudomonas osteochondritis and septic arthritis following nail puncture wound to the foot was diagnosed in 77 children aged 18 mo-19 y (77 and 17 cases, respectively). The syndromes were found in children with a history of wearing tennis shoes (70 cases), other shoes (5), and no shoes (2). All cases had surgical debridement of the infected cartilage or bone and drainage of infected joints. Pseudomonas aeruginosa was isolated in 38 cases and in conjunction with Staphylococcus aureus in 18. Anti Pseudomonas antibiotics were initiated in all cases before surgical exploration; the average duration of treatment was 7.5 +/- 1.2 d postoperatively. Patient follow-up was 5.2 +/- 3.4 y (median, 4.8 y; range, 3 mo-10 y). Two relapses occurred; both patients had a previously undetected septic arthritis. These data suggest that with aggressive surgical management, Pseudomonas osteochondritis and septic arthritis can be treated effectively with postoperative antibiotics for 7 d. PMID- 2571648 TI - [Gamma-glutamyltranspeptidase from Escherichia coli K-12]. PMID- 2571649 TI - [A study on the inhibition of non-specific neutral proteinase by gold salt. Part 1. Gold sodium thiomalate]. AB - The inhibitory effect of gold sodium thiomalate on non-specific neutral proteinase (EC 3.4.24.4) was investigated in vitro, in order to clarify the suppressive mechanism of the inflammatory process of joints by gold salt. The neutral proteinase was inhibited time- and concentration-dependently by gold salt. When the enzyme was incubated with 1 x 10(-2)M gold sodium thiomalate for 60 minutes, almost 100% of the activity was lost. Even at a lower concentration (1 x 10(-5)M, gold sodium thiomalate inhibited the enzyme time-dependently after longer incubation (3 or 9 hours). However, the same concentration of gold sodium thiomalate previously incubated with alpha-casein did not inhibit it. The results obtained here suggest that gold sodium thiomalate may inhibit the enzyme by binding it. The delay of neutral proteinase inhibition by gold sodium thiomalate seemed to be dependent on the concentration of gold salt. PMID- 2571650 TI - [Diagnosis and follow-up of ovarian cancer by a combination assay of serum sialyl SSEA-1 antigen and CA125 levels]. AB - We used a combination assay of serum sialyl SSEA-1 antigen (SLX) and CA125 levels, and evaluated the clinical usefulness of this technique for a diagnosis of ovarian cancer and follow-up of the patient with ovarian cancer. In 28 patients with ovarian tumors, the sera of 8 (66.7%) of 12 with ovarian cancer and 5 (71.4%) of the 7 with endometriosis (endometrial cyst) were positive for both SLX and CA125, but serum SLX level was 50 U/ml or less in all these 5 SLX-and CA125 positive patients with endometriosis. The sera of all 9 patients with benign ovarian tumor were negative for both tumor markers. No patient with endometriosis was negative for both markers. The diagnostic accuracy (true positive rate X true negative rate) of the combination assay for ovarian cancer was 50.3% when the cut-off value of the serum SLX was 38 U/ml but improved to 81.8% when the value was set at 50 U/ml. From the above observations, a combination assay of serum SLX and CA125 is promising method for the differential diagnosis of malignant and benign ovarian tumors. Our results also suggest that to improve the diagnostic accuracy, the cut-off value of the serum SLX level should be 50 U/ml for ovarian tumors alone. We found following-up two cases of ovarian cancer that the serum SLX level is not affected by the ascites and inflammation. We expect that this combination assay of serum SLX and serum CA125 will be beneficial for diagnosis and follow-up of ovarian cancer. PMID- 2571651 TI - [A case of type 1 multiple endocrine neoplasia with chondrosarcoma in the right lung]. PMID- 2571652 TI - [A case of concurrent occurrence of acute renal failure and nephrotic syndrome induced by alclofenac]. PMID- 2571653 TI - [Effects of different dosages of roxatidine on 24 hrs intragastric pH]. PMID- 2571654 TI - Mutations induced by 60Co gamma-irradiation in double-stranded M13 bacteriophage DNA in nitrous-oxide saturated solutions are characterized by a high specificity. AB - Irradiation of double-stranded M13 mp10 DNA in a diluted aqueous solution under N2O leads to a very specific mutation spectrum. Fifteen of 28 mutations induced in a 144 base pair (bp) target are C/G to G/C transversions, the other five bp substitutions are C/G to A/T transversions. Six mutations were single bp deletions, one is a large deletion of 180 bp and one is a 10 bp duplication which is probably from spontaneous origin. The mutations are not randomly distributed throughout the 144 bp mutation target but concentrated around two sites. The differences and similarities with the radiation-induced mutation spectrum previously obtained under oxygen are discussed. PMID- 2571655 TI - Transformation of lung cells from inhalation of radon daughters in dwellings: a preliminary study. AB - There is now world-wide concern for the quantification of lung cancer risk due to indoor radon, but the recent estimates are based on epidemiological studies of miners alone. The present attempt is a preliminary study of the alteration of lung stem cells irradiated by alpha particles emitted by radon daughters. Local energy deposited has been calculated for alpha particles, emitted from radon daughters lining the mucous layer in the respiratory tracts. This calculation has then been followed by dose evaluation and estimation of transformation of lung cells as a function of age. Mean life span of the stem cells was varied between 5 and 45 years to simulate living conditions in different environments. The cumulative fraction of transformed cells after 40 and 70 years has been calculated for radon concentration in the range 23-230 Bq/m3. Increase of the fraction of transformed cells with radon concentration was exponential. It has been assumed that causes other than radiation increase the rate of cell death of mature and stem lung cells, and hence the turnover of stem cells to replace them. It is concluded that the rate of transformation of cells is small for low radon concentration even late in age for non-polluted environments. For radon concentrations of 50 and 100 Bq/m3 the fractions of transformed cells are 0.2 and 6 per cent, respectively for an exposure time of 70 years. PMID- 2571656 TI - Protein-DNA crosslinks induced by primary and secondary radicals. AB - Double-stranded DNA from calf thymus was irradiated in the presence of bovine serum albumin (BSA) at a ratio of 1:10 by mass under air, N2 or N2O, with and without ethanol, tert-butanol or isopropanol in 10(-2) mol dm-3 sodium phosphate buffer, pH 7.0. The irradiated solutions were analysed by a nitrocellulose filter assay and by high-performance liquid gel chromatography. The results show that OH. radicals as well as hydrated electrons induce protein-DNA crosslinks. Further, alcohol radicals react with BSA producing protein radicals which become crosslinked to DNA. If OH radicals are scavenged by alcohol, strand breakage of DNA does not occur. PMID- 2571657 TI - CHEF electrophoresis, a sensitive technique for the determination of DNA double strand breaks. AB - It has been demonstrated that clamped homogeneous electrical field (CHEF) electrophoresis is a suitable method for the determination of DNA double-strand breaks in Chinese hamster ovary (CHO) cells. It allows the separation of DNA molecules up to 10 Mbp. The fraction of DNA fragments of this size is correlated with the number of radiation induced double-strand breaks. The resolution limit of the technique is equivalent to the effect of about 1 Gy (gamma-rays). Double strand break repair was monitored after irradiation with Co-60 gamma rays and the repair time constant determined to t1/2 = 30-35 min. In combination with the detection of DNA by fluorescence, CHEF electrophoresis provides an easy and sensitive method for the determination of double-strand break repair which does not require the radioactive labelling of cells. PMID- 2571658 TI - The lack of long-term recovery and reirradiation tolerance in the mouse kidney. AB - Mouse kidneys were bilaterally irradiated with X-ray doses of either 6 or 10 Gy (equivalent to approximately 30 and 70 per cent of full tolerance respectively). After an interval of 2 or 26 weeks the mice were retreated with a range of test doses given as single or fractionated irradiation schedules. Functional kidney damage was measured (using clearance of [51Cr]EDTA) before retreatment and at monthly intervals up to 1 year after retreatment. Reirradiation tolerance was assessed from dose-response curves for renal damage in retreated mice compared with that in age matched controls which received only the second treatment. Damage from the initial radiation doses progressed, leading to a decreased reirradiation tolerance with time. There was no evidence for any recovery from functional damage in the interval between 2 and 26 weeks. These studies would strongly suggest that reirradiation of a previously irradiated kidney (even after low initial doses below tolerance) is likely to lead to severe renal damage. Despite the poor reirradiation tolerance at 26 weeks, there was no reduction in the capacity for repair of sublethal injury when fractionated irradiation was given to previously irradiated mice compared with controls. PMID- 2571659 TI - The physiological state as a modifier of radiation-induced cytotoxicity in heterogeneous murine tumor cells growing in vitro. AB - The oxic radiation response (cytotoxicity) of two heterogeneous murine tumor-cell lines cultured in vitro was studied as a function of the cell's physiological state at the time of X-irradiation. The proliferating (P) 66 and 67 cells displayed equal radiosensitivities; however, the quiescent (Q) cells were considerably more radiosensitive than the P cells, and the 66Q cells were even more radiosensitive than the 67Q cells. Also, the 66Q cells continued to proliferate slowly with about 85 per cent in the G1 phase and 10 per cent in the S phase, while the 67 Q cells displayed a more complete G1 arrest (92-95 per cent). A detailed analysis of the metabolic status vs cell-cycle age (i.e. G1 vs S phase) indicated that the cell-cycle age was the predominant factor influencing radiation-induced cytotoxicity in 67 cells. The data also showed that in the plateau phase Q-cell cultures, pH and cell contact were not influencing factors and that the increased radiosensitivity of the Q cells could not be explained on the basis of energy deprivation. Moreover, the 66Q, but not the 67Q cells displayed an increased sensitivity in addition to that caused by the predominant cell-cycle age shift. This extra increase in radiosensitivity is of unknown metabolic origin, but could be related to cellular membrane fragility in the stressed 66Q cells since this extra component of Q-cell radiosensitivity was reduced both by refeeding (metabolic activation) 4 h before X-irradiation and by delayed plating while incubating the cells in Q medium at 37 degrees C after X irradiation. PMID- 2571661 TI - The cytological and histological effects of photoperiodism and X-irradiation on the testes of the Syrian hamster. AB - The radiosensitivity of meiotic stem cells from photoregressed testes of the Syrian hamster was studied after in vivo irradiation with 2.67 Gy (1.77 Gy/min). The testes of Expt I animals were irradiated at the time of transfer from short days (11 weeks at 6L:18D (6 hours light: 18 hours) darkness) to long days (14L:10D) and sampled at weekly intervals between 5 and 10 weeks and at 15 weeks postirradiation. The Ext II animals received the same X-ray dose, 2, 4, 6, 8 weeks after the transfer to long days and were sampled 10 weeks postirradiation. In Expt I X-irradiation delayed the recovery of testis structure and function by 4 weeks. Animals in Expt II were homogeneous with regard to testicular weight, sperm count, tubule diameter and the level of pachytene, MI and MII damage. The incidence of structural chromosomal changes in primary spermatocytes at pachytene (synaptonemal complexes) and MI and MII was determined. In Exp I and II and data from X-irradiated testes (2.6 Gy) not exposed to short days (Control B, see Cawood and Breckon 1983), these cell types show a difference in sensitivity to the induction of structural damage. The lowest level of pachytene damage was in Expt II, and the ratio of the number of pachytene to MI cells showing structural change was 1.23:1 (P = 0.41), not significantly different from 1:1. In Expt I the pachytene, MI structural change ratio was 1.60:1 (P = 0.010), significantly different from 1:1. The control B ratio was 1.85:1 (P = 0.0037) which was significantly different from 1:1. The preliminary results indicate that testes fully regressed to the 'stem' cell level by exposure to short day length have a similar radiosensitivity to normal (non-regressed) testes. However, testes irradiated during the recovery phase from photoperiod-induced regression have increased sensitivity to radiation damage. The relative testis weight was reduced to a mean of 1.25 g by the sequential X-ray treatment--this may indicate that the effects of the X-rays (3 Gy) on a recovering photoregressed testis are independent of the time of the X-ray treatment during the recovery period. Recovery of tubule diameter is not perturbed and may be slightly increased, with an associated elevation of MI-MII activity--but a low spermatogonia count.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2571660 TI - Radioprotection of C3H mice by recombinant human interleukin-1 alpha. AB - High doses of rHIL-1 protected C3H/HeN mice against lethal radiation damage. The optimum time of radioprotection was found when this cytokine was administered at 4 h and at 20 h before irradiation. The dose reduction factors when the IL-1 was administered at those times were about 1.30. The radioprotection at the optimum times was also observed in the cell kinetic behaviour of GM-CFC in the bone marrow. In previous studies it was reported that less radioprotection was found in C3H/HeN mice pretreated with IL-1. This may not be the case with a higher dose of IL-1. PMID- 2571662 TI - The efficacies of pure LICAM(C) and DTPA on the retention of plutonium-238 and americium-241 in rats after their inhalation as nitrate and intravenous injection as citrate. AB - The pure carboxylated catechoyl amide LICAM(C) and the calcium and zinc salts of diethylenetriaminepenta-acetic acid (DTPA), were tested for efficacy for removing 238Pu and 241Am from rats after inhalation of the nitrate or intravenous injection of the citrate. The results were compared with the efficacy of methylated LICAM(C) used in previous experiments. It was shown that: (1) after inhalation of 238Pu nitrate, DTPA was far superior to pure LICAM(C); (2) after intravenous injection of 238Pu citrate, the infusion of DTPA plus LICAM(C) was only marginally more effective than DTPA alone; and (3) after inhalation or intravenous injection of 238Pu plus 241Am, the efficacy of pure LICAM(C) was only marginally more effective than the methylated form and neither form was effective for the decorporation of 241Am. It was concluded that DTPA, at present, remains the chelating agent of choice for treating persons accidentally contaminated with transportable forms of Pu and Am. PMID- 2571663 TI - Use of serum gamma glutamyl transpeptidase to differentiate between extrahepatic biliary atresia and neonatal hepatitis. AB - The differentiation between extrahepatic biliary atresia (EHBA) and neonatal hepatitis (NH) was studied in 53 Thai infants (39 males, 14 females) with obstructive cholangiopathy at the Department of Pediatrics, Siriraj hospital. Prevalence of EHBA was 25 per cent (13 of 15) overall. A higher prevalence of EHBA was found in females (8 of 13) while a higher prevalence of NH occurred in males (34 of 40). No difference was demonstrated in serum conjugated and total bilirubin, SGOT or alkaline phosphatase between EHBA and NH groups. The EHBA group had a significantly higher mean serum GGTP (184 +/- 77 IU/L) than the NH group (58 +/- 40 IU/L). The diagnostic usefulness of GGTP was confirmed, but variation in laboratory measurements could limit its generalizability. PMID- 2571664 TI - Abrupt change from a beta-adrenoceptor blocking drug to enalapril in hypertension. AB - The haemodynamic and hormonal effects of an elective change of antihypertensive therapy from a beta-adrenoceptor blocking drug to a converting enzyme inhibitor, enalapril, were monitored in 12 hypertensive in-patients (WHO I). Blood pressure and heart rate were determined every 2-4 h using an automatic sphygmomanometer during an abrupt cessation of the previous beta-adrenoceptor blocking drug and commencement of treatment with enalapril 20 mg o.d. 12 h later. Mean blood pressure values at rest and during the hand grip test were lower when on enalapril, but heart rate was significantly higher, and three patients suffered from palpitations during the change. The change resulted in an improvement in cardiac function both at rest and during isometric work, as shown by echocardiography. A rapid decrease in plasma angiotensin converting enzyme (ACE) activity and an increase in renin activity were also seen after the change, while plasma levels of atrial natriuretic peptide (ANP) decreased towards normal values. The results suggest that an abrupt change from a chronic beta adrenoceptor blocking drug to enalapril is safe, feasible and is likely to produce favourable haemodynamic and hormonal effects in hypertensive patients. PMID- 2571665 TI - Expansion of natural killer cells but not T cells in human interleukin 2/interleukin 2 receptor (Tac) transgenic mice. AB - Transgenic mice expressing both human IL-2 and the L chain of IL-2-R constitutively had an unusual expansion of Thy-1+/CD3-4-8- large granular lymphocytes, which bore the elevated NK activity. Unexpectedly, the transgenic mice had neither T cell expansion nor autoreactive antibodies. The increase in number and activity of NK cells seems to be responsible for both the severe interstitial pneumonia and lymphocyte depletion in the spleen that we found in these transgenic mice. In addition, we found the selective loss of Purkinje cells in the cerebellum of the mice, which gave rise to their disturbed gait. All the transgenic mice died by 4 wk of age. PMID- 2571666 TI - Infection of monocyte-derived macrophages with human immunodeficiency virus type 1 (HIV-1). Monocyte-tropic and lymphocyte-tropic strains of HIV-1 show distinctive patterns of replication in a panel of cell types. AB - To characterize the host range of different strains of HIV-1, we have used four types of cells, primary monocyte-derived macrophages (MDM), primary PBL, a promonocyte cell line (U937), and a CD4+ T cell line (SUP-T1). These cells were infected with three prototype strains of HIV-1, a putative lymphocyte-tropic strain (IIIB), and two putative monocyte-tropic strains (SF162 and DV). Infections were monitored by assays for infectious virus, for cell-free and cell associated viral antigen (p24), and for the proportion of cells infected by immunohistochemical staining. It was concluded that: (a) the use of four different cell types provides a useful biological matrix for distinguishing the tropism of different strains of HIV-1; this matrix yields more information than the infection of any single cell type. (b) A monocyte-tropic strain of HIV-1, such as strain SF162, shows a reciprocal host range when compared with a lymphocyte-tropic strain such as IIIB; strain SF162 replicates well in primary MDM but not in U937 or SUP-T1 cells, while strain IIIB replicates well in both U937 and SUP-T1 cells but not in MDM. (c) Both lymphocyte-tropic and monocyte tropic strains of HIV-1 replicate well in PBL. (d) The promonocyte cell line, U937, and the T cell line, SUP-T1, differ markedly from primary cells, such as MDM and PBL, in their ability to support the replication of different strains of HIV-1; these cell lines cannot be used as surrogates for primary cells in host range studies of HIV-1 strains. PMID- 2571667 TI - Insertion/deletion-related polymorphisms in the human T cell receptor beta gene complex. AB - Insertion/deletion related polymorphisms (IDRP) involving stretches of 15-30 kb within the human TCR-beta gene complex were revealed by pulse-field gel electrophoresis. Two independent IDRP systems were detected by analysis of Sfi I- and Sal I-digested human DNA samples using probes for TCR C and V region gene segments. The allelic nature of these systems was verified in family studies, and mapping data allowed localization of one area of insertion/deletion among the V gene segments and the other near the C region genes. All but one of 50 individuals tested could be typed for the two allelic systems, and gene frequencies for the two allelic forms were 0.37/0.61 and 0.46/0.54, indicating that these polymorphisms are widespread. PMID- 2571668 TI - Chlamydial disease pathogenesis. The 57-kD chlamydial hypersensitivity antigen is a stress response protein. AB - Chlamydia trachomatis infection of humans is commonly a localized inflammation that can result in infertility, blindness, and perhaps arthritis. The pathogenic process(es) that cause these sequelae are thought to be immunological. A 57-kD protein that is common among Chlamydia elicits ocular inflammation when introduced onto the conjunctivae of guinea pigs or nonhuman primates previously sensitized by chlamydial infection. This protein is thought to mediate the immunopathology that follows chlamydial infection. To more thoroughly characterize this chlamydial component, we cloned its gene from a C. psittaci strain and identified a particular recombinant that produced the 57-kD polypeptide. The recombinant gene product was immunoreactive with a monospecific anti-57-kD serum, and elicited an ocular inflammation similar to that produced by the 57-kD antigen isolated from chlamydiae. Sequencing identified two ORFs that encode polypeptides of 11.2 and 58.1 kD and are co-transcribed. These two polypeptides show homology with Escherichia coli groE and Coxiella burnetii htp heat-shock proteins. Striking homology (greater than 50%) was found between the 57-kD protein and the HtpB, GroEL, 65-k Mycobacterium tuberculosis and Hsp60 proteins. Thus, the 57-kD chlamydial protein, previously implicated as mediating a deleterious immunologic response to chlamydial infections, is a stress-induced protein similar to those that occur universally in both prokaryotic and eukaryotic organisms. PMID- 2571669 TI - Analysis of restriction fragment length polymorphism in lymphokine genes of normal and autoimmune mice. AB - Analysis of RFLP has been employed in lymphokine genes of autoimmune and normal mice. No polymorphism could be detected in the loci containing IL-2, IL-2 receptor, IL-5, and IFN-gamma in NZB, NZW, BxSB, and MRL/lpr mice when compared with normal mice. Allelic forms were identified in the IL-1 alpha gene of BALB/c and in the IL-4 gene of NZW. The frequency of the Bam HI RFLP in the TNF-alpha gene of NZW which has been proposed to be associated with the development of autoimmune disease in (NZB x NZW)F1 mice has been analyzed in a number of different inbred strains and in wild mice. Since the same allele is inherited in most autoimmune, healthy laboratory and wild mice the TNF-alpha gene does not seem to be one of the causal agents that contributes to the development of autoimmunity in (NZB x NZW)F1 mice. PMID- 2571670 TI - Activation of peripheral blood T cells via the p75 interleukin 2 receptor. AB - By using mAb and flow cytometry, a constitutive expression of the p75 IL-2R was revealed in human peripheral blood CD8+ T cells and TCR delta-1+ T cells as well as in CD16+ NK cells. Anti-p75 IL-2R mAb almost completely inhibited the induction of cytolytic activity in these T cells by brief exposure to IL-2, as estimated by anti-TCR/CD3 mAb-targeted cytotoxicity. While anti-p55 IL-2R mAb alone inhibited the response only modestly, maximal inhibition was achieved by combining both anti-p55 and anti-p75 IL-2R mAbs. These results indicate that the p75 IL-2R constitutively expressed on peripheral blood CD8+ T cells and TCR delta 1+ T cells is predominantly responsible for the direct activation of these cells by IL-2. PMID- 2571671 TI - Clinical significance of urinary N-acetyl-beta-D-glucosaminidase and alanine aminopeptidase. AB - The excretion of urinary N-acetyl-beta-D-glucosaminidase (NAG) and Alanine Aminopeptidase (AAP) increases with renal damage. The variation in enzyme activity due to the fluctuation of urine flow rate could almost be eliminated by expressing it as the ratio of enzyme activity to urinary creatinine concentration. The urinary enzyme activities increased not only by tubular damage but also by the reduced creatinine clearance due to glomerular injury. The normal reference values of NAG were 2.84 +/- 2.50U/g creatinine for 24-hour urine and 3.23 +/- 2.76U/g creatinine for random urine. The normal reference values of AAP were 9.71 +/- 6.68U/g creatinine for both 24-hour urine and random urine. Although 128 patients were with abnormal enzymuria, only 39.1%, 52.3%, 83.6%, and 86.7% of the patients were with abnormal serum creatinine, serum urea N, creatinine clearance, and urine protein, respectively. In view of the sensitivity, the determinations of urinary NAG and AAP are suitable for early detection of renal diseases. PMID- 2571672 TI - Plasma homovanillic acid in schizophrenics: supportive evidence for the two subtype hypothesis. AB - Plasma levels of homovanillic acid (pHVA), a major metabolite of dopamine (DA), were measured in a group of 51 schizophrenic inpatients before and during 6 weeks of neuroleptic treatment. Steady-state plasma drug concentrations were monitored in parallel with pHVA. Good responders (n = 22) had higher pretreatment pHVA levels as compared to poor responders (n = 22). Differential pHVA changes during neuroleptic treatment were also found between each group. The two groups did not differ significantly in terms of age, duration of illness, severity of presenting symptoms, neuroleptic, dose, or plasma drug concentration. Two hypothetical subtypes in the group of schizophrenics were proposed. PMID- 2571674 TI - Bovine brain endothelial cells express tight junctions and monoamine oxidase activity in long-term culture. AB - The passage of substances across the blood-brain barrier is regulated by cerebral capillaries which possess certain distinctly different morphological and enzymatic properties compared to capillaries of other organs. Investigations of the functional characteristics of brain capillaries have been facilitated by the use of cultured brain endothelial cells, but in most studies a number of characteristics of the in vivo system are lost. To provide an in vitro system for studies of brain capillary functions, we developed a method of isolating and producing a large number of bovine brain capillary endothelial cells. These cells, absolutely free of pericyte contamination, are subcultured, at the split ratio of 1:20 (20-fold increase of the cultured surface), with no apparent changes in cell morphology up to the fiftieth generation (10 passages). Retention of endothelial-specific characteristics (factor VIII-related antigen, angiotensin converting enzyme, and nonthrombogenic surface) is shown for brain capillary derived endothelial cells up to passage 10, even after frozen storage at passage 3. Furthermore, we showed that bovine brain capillary endothelial cells retain, up to the fiftieth generation, some of the characteristics of the blood-brain barrier: occurrence of tight junctions, paucity of pinocytotic vesicles, and monoamine oxidase activity. PMID- 2571675 TI - ATP release, adenosine formation, and modulation of dynorphin and glutamic acid release by adenosine analogues in rat hippocampal mossy fiber synaptosomes. AB - Using a hippocampal subcellular fraction enriched in mossy fiber synaptosomes, evidence was obtained indicating that adenosine derived from a presynaptic pool of ATP may modulate the release of prodynorphin-derived peptides. and glutamic acid from mossy fiber terminals. Synaptosomal ATP was released in a Ca2+ dependent manner by K+-induced depolarization. The rapid hydrolysis of extracellular [14C]ATP in the presence of intact mossy fiber synaptosomes resulted in the production of [14C]adenosine. Micromolar concentrations of a stable adenosine analogue, 2-chloroadenosine, inhibited the K+-stimulated release of both dynorphin B and dynorphin A(1-8). 2-Chloroadenosine failed to suppress the evoked release of glutamic acid, measured in these same superfusates, unless the mossy fiber synaptosomes were pretreated with D-aspartic acid to deplete the cytosolic, Ca2+-independent, pool of this acidic amino acid. In synaptosomes pretreated in this manner, release of the remaining Ca2+-dependent pool of glutamic acid was significantly inhibited by NiCl2, 2-chloroadenosine, 5'-N ethylcarboxamidoadenosine, cyclohexyladenosine, and R(-)-N6(2 phenylisopropyl)adenosine, but not by ATP. 2-Chloroadenosine-induced inhibition was reversed when the external CaCl2 concentration was raised from 1.8 mM to 6 mM. 8-Phenyltheophylline, an adenosine receptor antagonist, effectively blocked the inhibitory effects of 2-chloroadenosine on mossy fiber synaptosomes and significantly enhanced the K+-evoked release of both glutamic acid and dynorphin A(1-8) when added alone to the superfusion medium. These results support the proposition that depolarized hippocampal mossy fiber synaptosomes release endogenous ATP and are capable of forming adenosine from extracellular ATP, and that endogenous adenosine may act at a presynaptic site to inhibit the further release of glutamic acid and the prodynorphin-derived peptides. PMID- 2571673 TI - [Mosquito bite allergies terminating as hemophagocytic histiocytosis: report of a case]. AB - A 21-year-old woman had suffered from repeated vasculitis and panniculitis with fever and chills after following mosquito bites since age 7. These manifestations were confirmed with a direct mosquito bite test during her admission to another hospital at age 17. The patient presented to our hospital with progressive dyspnea, productive cough and intermittent fever for one week. In addition to the bilateral infiltrative lesions on chest roentgenography, hepatosplenomegaly with an abnormal liver function test, pancytopenia, and elevated IgE were also detected. Blood and sputum cultures grew no microorganisms. Epstein-Barr virus IgM, Cytomegalovirus-IgM and Mycoplasma pneumonia antibodies were all negative. Bone marrow aspiration and biopsy revealed histiocytosis with hemophagocytosis. No atypical histiocyte was found. The patient was put on dexamethasone treatment with improvement. Unfortunately, symptoms relapsed two weeks later. A repeated bone marrow aspiration and biopsy revealed a picture similar to the previous one. Despite antibiotic administration and ventilator support, a rapidly deteriorated course terminated in the patient's death by respiratory failure. Mosquito bite allergies were reported to be associated with malignant histiocytosis in Japan. However, atypical histiocytes were not found in our case upon repeated bone marrow aspirations, biopsies and skin biopsies. Mature histiocytes with hemophagocytosis were prominent instead. Reactive histiocytosis was thus favored rather than malignant histiocytosis. Although opportunistic infection cannot be excluded, we propose that a mosquito bite allergy with consequent histiocytic activation by antigen, immune complex or IgE is the possible pathogenetic mechanism for hemophagocytic histiocytosis in this patient. PMID- 2571677 TI - Isolation of nerve terminals from crustacean muscle. AB - A method for the isolation of gamma-aminobutyric acidergic (GABAergic) and glutamatergic terminals from crustacean muscle was developed, using differential centrifugation and sucrose density gradient centrifugation. Individual fractions were assessed using a variety of markers. One fraction was isolated which showed 40-fold purification of glutamate decarboxylase with a yield of 12%. This fraction was enriched in GABA, glutamate, glutamate dehydrogenase, and 5' nucleotidase, but not in NADPH cytochrome c reductase. This fraction possessed an uptake system for GABA and glutamate with apparent kinetic constants of Km = 50 microM, Vmax = 250 pmol/min/mg of protein and Km = 183 microM, Vmax = 219 pmol/min/mg of protein, respectively. Electron microscopy showed nerve terminal profiles and a heterogeneous population of membrane vesicles. This fraction contained 3.4 nmol ATP/mg of protein which was stable for 30 min at 12 degrees C, and was also able to synthesise ATP from exogenous adenosine. The terminals released labelled GABA and glutamate in a Ca2+-dependent fashion on depolarisation. No release of ATP was detected. It is concluded that viable nerve terminals have been isolated which could be used as model systems for the study of GABAergic and glutamatergic neurochemistry. PMID- 2571676 TI - A comparison of the distribution of neurotransmitters in brain regions of the rat and guinea-pig using a chemiluminescent method and HPLC with electrochemical detection. AB - Six brain areas of rats and guinea-pigs, killed by microwave irradiation, were used for the concomitant measurement of the levels and regional distribution of cholinergic, biogenic amine, and amino acid neurotransmitters and metabolites. Acetylcholine (ACh) and choline (Ch) were quantified by chemiluminescence; noradrenaline (NA), dopamine (DA), 5-hydroxytryptamine (5-HT), and their metabolites by HPLC with electrochemical detection (HPLC-EC); and six putative amino acid neurotransmitters by HPLC-EC following derivatisation. The levels and regional distribution of these transmitters and their metabolites in the rat were similar to those reported in previous studies, except that biogenic amine transmitter levels were higher and metabolite concentrations were lower. The guinea-pig showed a similar regional distribution, but the absolute levels of ACh were lower in striatum and higher in hippocampus, midbrain-hypothalamus, and medulla-pons. In all areas, the levels of Ch were higher and those of NA, 5-HT, and taurine were lower than in the rat. The most marked differences between the rat and guinea-pig were in the relative proportion of DA metabolites and 5-HT turnover, as estimated by metabolite/transmitter ratios. This study can be used as a basis for a comprehensive understanding of the central effects of drugs on the major neurotransmitter systems. PMID- 2571678 TI - Increased intracellular cyclic AMP differentially modulates nerve growth factor induction of three neuronal recognition molecules involved in neurite outgrowth. AB - The relative expression of the immunoglobulin superfamily members Thy-1 and L1 and the neural cell adhesion molecule (N-CAM) in PC12 cells grown in the presence of nerve growth factor (NGF), cholera toxin, or both has been quantified. Whereas NGF treatment induced increases in the cell surface expression of all three glycoproteins, treatment with cholera toxin resulted in the specific induction of L1. During the first few days of culture, cholera toxin acted synergistically with NGF to promote increases in neuritic outgrowth and the synthesis and cell surface accumulation of the 140- and 180-kilodalton subunits of N-CAM. In contrast, over the same period of culture, cholera toxin inhibited the NGF induction of Thy-1 and L1. Over longer periods of culture (3-5 days), cholera toxin inhibited the NGF induction of N-CAM and neurite outgrowth. A similar pattern of synergistic and inhibitory responses was observed when differentiation was induced by fibroblast growth factor (FGF) rather than NGF or when cholera toxin was replaced with forskolin. These data suggest that intracellular cyclic AMP can differentially modulate cell surface glycoprotein expression induced by either NGF or FGF. Of the three cell surface glycoproteins we have studied, temporal changes in N-CAM expression correlate best with the morphological differentiation status of PC12 cells. PMID- 2571679 TI - 5' flanking DNA sequences direct cell-specific expression of rat tyrosine hydroxylase. AB - Tyrosine hydroxylase (TH) is selectively expressed in catecholaminergic neurons and in chromaffin cells of the adrenal medulla. Constructs in which 5' flanking sequences of the rat TH gene directed expression of bacterial chloramphenicol acetyltransferase (CAT) were transfected into cell lines and assayed for transient expression of CAT. In most nonexpressing cell lines, CAT levels were less than 5% of that found in a TH-positive pheochromocytoma line (PC8b). In two lines described here, a rat anterior pituitary cell line (GH4) and a rat fibroblast line (Fr3T3), CAT expression reached 12 and 20%, respectively, of the PC8b level. Greater than 90% of the PC8b activity was lost when sequences between -212 and -187 (in relation to the transcriptional initiation site) were deleted. Further deletions that removed the cyclic AMP response element (CRE) (-45) and the TATA box at -29 reduced transcriptional activity to background in all three lines. These data suggest that 212 nucleotides of the 5' sequence are sufficient for pheochromocytoma expression and that information between -212 and -187, which includes an AP1 site (-206 to -200), is essential for full transcriptional activity. In addition, sites for other protein transcription factors (AP2, POU/Oct, SP1, and CRE) reside between -221 and -38 and are largely conserved between the human and rat gene. PMID- 2571680 TI - Fatigue syndromes: a comparison of chronic "postviral" fatigue with neuromuscular and affective disorders. AB - Patients (n = 47) presenting to a neurological centre with unexplained chronic "postviral" fatigue (CFS) were studied prospectively. Controls were patients with peripheral fatiguing neuromuscular diseases and inpatients with major depression in a psychiatric hospital. Seventy-two percent of the CFS patients were cases of psychiatric disorder, using criteria that excluded fatigue as a symptom, compared with 36% of the neuromuscular group. There was no difference in subjective complaints of physical fatigue between all groups. Mental fatigue and fatigability was equally common in CFS and affective patients, but only occurred in those neuromuscular patients who were also cases of psychiatric disorder. Overall, the CFS patients more closely resembled the affective than the neuromuscular patients. Attribution of symptoms to physical rather than psychological causes was the principal difference between matched CFS and psychiatric controls. The symptoms of "postviral" fatigue had little ability to discriminate between CFS and affective disorder. The fatigue in CFS appeared central in origin, suggesting it is not primarily a neuromuscular illness. The implications for research and treatment of chronic fatigue are discussed. PMID- 2571682 TI - Almitrine-induced peripheral neuropathy and weight loss. PMID- 2571681 TI - Circulating CD4+CD8+ cells in myasthenia gravis: supplementary immunological parameter for long-term prognosis. AB - Twenty patients with myasthenia gravis (MG) were studied prospectively for up to 5 years after thymectomy, in order to clarify the relationships between disease severity, anti-acetylcholine receptor antibody (anti-AChR) titres, proportions of circulating CD4+CD8+ cells (CD4+CD8+ cell level) and major lymphocyte subsets. The CD4+CD8+ cell levels were closely related to the clinical change within 1 year after surgery in 8 patients who showed a preoperative elevation in the cell levels. This group of patients consisted of six thymomatous and two non thymomatous patients; the latter were both negative for anti-AChR. The anti-AChR titres generally changed in parallel with the clinical state in 9 of the 16 patients who were followed up for more than a year after thymectomy, and the CD4+CD8+ cell levels were useful in predicting the clinical course in 6 of the above 9 patients and 3 other patients, including antibody-negative cases. The present study suggests that the CD4+CD8+ cell levels may serve as an indicator for long-term prognosis of MG. PMID- 2571683 TI - A muscle-derived factor(s) induces expression of a catecholamine phenotype in neurons of cultured rat cerebral cortex. AB - We sought to determine the source of the signal(s) that promotes expression of the catecholamine (CA) enzyme tyrosine hydroxylase (TH) in cultured neurons of embryonic rat cerebral cortex, a tissue which is not thought to contain CA cells in vivo. Cortical neurons were cultured with their non-neuronal constituents and 48 hr later immunostained for TH. Fibroblasts or glia had no effects, however, blood vessels increased the numbers of TH neurons nearly 4-fold. Coculture with either perinatal aorta, skeletal or cardiac muscle, clonal muscle cell lines 1440 (smooth) and L6 (skeletal), conditioned media from L6 cells, or a soluble extract of L6 cells increased the number of TH neurons up to 20-fold. The induction of TH by muscle extract was (1) dose dependent; (2) paralleled by a proportional increase in the steady-state levels of TH mRNA; (3) greatly reduced by the RNA synthesis inhibitor alpha-amanitin or the protein synthesis inhibitor cycloheximide; and (4) unassociated with change in the survival of neurons in culture. The response was not replicated by treatment with other established neurotrophic substances, including NGF, EGF, FGF, PDGF, neuroleukin, insulin, pyruvate, KCI, adenosine, or inosine. We conclude that muscle contains a potentially novel substance, muscle-derived differentiation factor (MDF) that promotes differentiation but not survival of neurons of cerebral cortex by de novo synthesis of TH mRNA and TH protein. Thus, neurons of the CNS, as in periphery, may undergo phenotypic interconversion in response to biologically derived molecules in their environment. PMID- 2571685 TI - The neurotoxicity of 1-methyl-4-phenylpyridinium in cultured cerebellar granule cells. AB - Cerebellar granule cells in enriched primary culture are susceptible to the neurotoxic effects of 1-methyl-4-phenylpyridinium (MPP+). Relatively high MPP+ concentrations are required to elicit neurotoxic effects at early culture times, but lower concentrations of MPP+ produce comparable neurotoxic effects at later culture times. Under identical culture conditions 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) is not neurotoxic. Preincubation with the glutamate uptake blockers, DL-threo-3-hydroxyaspartic acid or dihydrokainate, or the dopaminergic uptake blocker mazindol, protects the granule cells from the cytotoxic effects of MPP+. Although MPTP is not neurotoxic in an enriched granule cell culture, in coculture with cerebellar astrocytes MPTP is toxic to granule cells, presumably because it is converted in astrocytes to MPP+. Cerebellar astrocytes remain confluent and viable. The addition of pargyline to the coculture abolishes the neurotoxicity consistent with a role of MAO B in bioactivation of MPTP. The concentration of MPP+ in the coculture medium (13 microM) was less than that required for the toxic effect in enriched neuronal cultures at earlier culture times, suggesting that an astroglial-neuronal interaction, perhaps by proximity, enhances the neurotoxicity of MPP+. These results might explain reported effects of MPTP on some cerebellar cells in mice. PMID- 2571684 TI - Multiple effects of phorbol esters in the rat spinal dorsal horn. AB - Spinal cord slice preparation and intracellular recording techniques were used to examine the effects of phorbol esters on the sodium- and calcium-dependent action potentials, the excitatory synaptic transmission, the basal (resting) and the dorsal root stimulation-evoked release of 9 endogenous amino acids, including glutamate and aspartate, and the responsiveness of the rat dorsal horn neurons to excitatory amino acids (glutamic, kainic, quisqualic, and N-methyl-D-aspartic). 4 beta-Phorbol-12, 13-dibutyrate and 4-beta-phorbol-12, 13-diacetate produced minor alterations in membrane potential and resistance, but they broadened the sodium dependent action potential and reduced the duration of the calcium-dependent action potential. In addition, phorbol esters caused a marked and long-lasting increase in the amplitude and the duration of excitatory postsynaptic potentials (EPSPs) evoked in dorsal horn neurons by orthodromic stimulation of a lumbar dorsal root. Phorbol esters produced a brief increase in the basal and electrically evoked release of endogenous excitatory (glutamic, aspartic) and inhibitory amino acids (glycine, GABA). In addition, the rates of release of alanine, serine, and threonine were also elevated. In the presence of TTX, phorbol esters selectively enhanced, in a reversible manner, the depolarizing responses of dorsal horn neurons to N-methyl-D-aspartic acid and L-glutamate but not the responses to kainic or quisqualic acids. The potentiation of the NMDA response was blocked by APV, a specific NMDA receptor antagonist. Thus, phorbol esters appear to enhance excitatory synaptic transmission in the rat spinal dorsal horn slice preparation by acting both at pre- and postsynaptic sites. Phorbol esters could potentiate excitatory synaptic transmission by acting predominantly at a postsynaptic site (NMDA receptor), since the duration of the increased responsiveness of dorsal horn neurons to glutamate and NMDA correlates better with the enhancement of EPSPs than with the increased release of the stimulation-evoked glutamate and aspartate. The increased release of endogenous amino acids is consistent with a presynaptic (terminal) site of action, but it could also be explained by enhanced interneuronal activity. Although our results suggest that in the rat spinal dorsal horn protein kinase C may have a role in controlling the release of putative excitatory and inhibitory neurotransmitters and may also be involved in the regulation of postsynaptic NMDA receptors, the identity of endogenous substance(s) participating in these effects is presently unknown. PMID- 2571686 TI - SCP-containing R20 neurons modulate respiratory pumping in Aplysia. AB - Respiratory pumping in Aplysia consists of transient, synchronous pumping actions of the gill, siphon, mantle shelf, and parapodia. This behavior has previously been shown to be driven by a network of coupled interneurons in the abdominal ganglion, the R25 and the L25 cells. We describe here a pair of electrically coupled cells, the R20 cells, which when active can initiate respiratory pumping or increase its spontaneous rate of occurrence. This action is mediated by a slow, long-lasting excitation of the endogenous burst mechanism of the cells in the R25/L25 network. The R20 cells, which are located in the abdominal ganglion, also make slow inhibitory connections to the RB cells and to the RG cells in that ganglion, and to the gill motoneurons in the branchial ganglion. The R20 cells are immunoreactive to SCPB, a molluscan neuropeptide. Biochemical purification studies demonstrate that each of the R20 cells synthesizes not only SCPB, but also SCPA, a closely related molecule known to be encoded by the same gene as SCPB. The R20 cells also synthesize in abundance several other low-molecular weight, methionine-containing peptides. The excitatory actions of the R20 cells on the R25/L25 network are mimicked by SCPA and SCPB. However, the inhibitory actions of the R20 cells on the RB cells, the RG cells, and on the cells of the branchial ganglion are not mimicked by the SCPs. Thus, the data support the hypothesis that the R20 cells release SCPA and SCPB and at least one other unidentified transmitter. PMID- 2571687 TI - Calcium current in cortical astrocytes: induction by cAMP and neurotransmitters and permissive effect of serum factors. AB - Voltage-dependent L-type calcium channels were induced in highly purified, cultured, cortical astrocytes by exposure to substances known to increase their intracellular cAMP: 8-bromo-cAMP, forskolin, isoproterenol, and vasoactive intestinal peptide. In untreated control cultures, L-type calcium currents were entirely absent. The induction of this calcium current was specific to cortical astrocytes and a closely related astrocyte type in white matter and did not occur in meningeal cells or oligodendrocytes. The ability of forskolin to induce L-type calcium current in astrocytes depended on previous culture of these cells in a permissive lot of serum for at least 48 hr. In addition, certain lots of sera caused expression of a voltage-dependent T-type calcium channel in untreated control cultures. Several possible mechanisms of calcium channel induction by cAMP are consistent with our data: modification of a silent channel already present in the membrane, indirect effect of cytoskeletal alteration, or insertion of channels from submembrane stores. If the environment in vivo is permissive, regulation of glial ion channels may be under neuronal control. PMID- 2571688 TI - Quantitative physiological characterization of a quinoxalinedione non-NMDA receptor antagonist. AB - The effects of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, or FG 9065) on excitatory amino acid responses in cultured neurons from rat hippocampus were studied using tight-seal whole-cell recording techniques. CNQX reduced the magnitude of peak inward currents produced by exogenously applied kainate, quisqualate, and N-methyl-D-aspartate (NMDA) with Ki's of 2.5, 3.5, and 96 microM, respectively. The antagonism was competitive against kainate and quisqualate, but noncompetitive against NMDA. Glycine markedly reduced CNQX antagonism of NMDA responses. The same recording technique using pairs of monosynaptically connected neurons demonstrated reversible diminution of excitatory postsynaptic potentials in 7 of 7 pairs, using CNQX at concentrations as low as 10 microM. CNQX applied alone did not evoke inward or outward currents at membrane potentials near the resting membrane potential and did not affect the current-voltage relationship at membrane potentials between -90 and -30 mV. These observations represent the first quantitative characterization of glutamate receptor antagonism by CNQX with respect to physiological rather than biochemical parameters and demonstrate that CNQX is far more potent and more selective than currently available non-NMDA antagonists. The results suggest that CNQX will be a useful pharmacologic tool for the study of synaptic transmission in a variety of systems in which glutamate or related excitatory amino acids are involved. PMID- 2571690 TI - Patterns of PRN analgesic drug administration in children following elective surgery. AB - Conducted chart review study of 114 4- to 14-year-olds hospitalized for elective surgery to investigate analgesic medication patterns. Correcting for body weight and different drug potencies, correlations were examined between analgesics, child age, painfulness of recovery, and seriousness of surgical procedure. Weight adjusted analgesics prescribed and delivered PRN were unrelated to painfulness of procedure and age. Expert ratings of the seriousness of anticipated sequelae were found to be a modest but significant predictor of analgesics. Results suggest that PRN prescription of analgesics in children essentially guarantees very low drug delivery without achieving individualized pain management. Possible interpretations and suggestions for research into effects of practitioner attributions of seriousness on clinical decision making are discussed, along with implications for other explanations of inadequate analgesic practices. PMID- 2571689 TI - Intracerebroventricular bethanechol chloride infusion in Alzheimer's disease. Results of a collaborative double-blind study. AB - The use of intracerebroventricular bethanechol chloride infusion in patients with Alzheimer's disease was first reported in 1984. An initial trial in four patients demonstrated the feasibility of this approach for cholinergic drug delivery to the brain, but objective improvement in cognitive function was not documented. A collaborative placebo-controlled double-blind crossover study has now been carried out in 49 patients with biopsy-documented Alzheimer's disease. The results demonstrate a statistical improvement in Mini-Mental State scores and significantly slower performance on Trails A testing during drug infusion. Other neuropsychological test scores were not similarly affected. The degree of improvement was not sufficient to justify further treatment of Alzheimer's disease patients by intracerebroventricular infusion of bethanechol chloride. The drug delivery system used in the study was well tolerated, with two irreversible complications in more than 50,000 patient days. PMID- 2571692 TI - Mechanistic toxicology: a radical perspective. PMID- 2571691 TI - [Potassium chlorazepate administered orally in alcoholic detoxification]. AB - 30 alcoholic patients were studied during a detoxification period of 14 days in the hospital. Variable doses (maximum 300 mg) of potassium clorazepate allowed to achieve an harmless and rather comfortable detoxification, evaluated through internationally used skates (SSA and HSCL). PMID- 2571693 TI - The effects of autoclaving on the physical properties and biological activity of parenteral heparin preparations. AB - Heparin ampoules have been autoclaved at 115 degrees C, 121 degrees C, 126 degrees C and 130 degrees C for time intervals up to 50 min and the biological potency and physicochemical integrity of the preparations assessed. The anticoagulant activity, determined with the APTT assay, did not change significantly for the autoclaved samples but a decrease was observed using the BP assay. Autoclaving was also associated with a depolymerization process, as confirmed by high performance liquid chromatography, and the formation of 'species' with an absorption in the UV spectrum. However, autoclaving had no detrimental effect on the integrity of the anionic sites on heparin. It is proposed that Maillard-type reactions are responsible for these observations and the results indicate that autoclaving could be used to sterilize parenteral heparin solutions. PMID- 2571694 TI - Calculation of punch displacement and work of powder compaction on a rotary tablet press. AB - To calculate the work of compaction during tableting it is necessary to have accurate values of force and punch displacement. The direct measurement of punch displacement on a rotary press is both costly and complicated but calculated displacements will be in considerable error unless deflections in the press during compression, are taken into account. By analysing the physical restraints imposed on the punches during tablet compression, an expression for punch displacement was derived. From preliminary measurements made on the table press of machine deflections and punch displacement under static conditions, the terms of this expression were evaluated for dynamic conditions. This analytic solution was then used to determine the true punch displacement and work of compaction from direct measurements of vertical force and turret position. PMID- 2571695 TI - The effect of gelatin grade and concentration on the migration of solutes into and through glycerogelatin gels. AB - The diffusion of 4-hydroxybenzoic acid and phenobarbitone through glycerogelatin gels was found to be independent of the type of gelatin used. Three types of gelatin, two acid-processed and one alkali-processed were studied, and the bulk viscosities of gels prepared from them was seen to vary considerably. However, the microviscosities of the gels, as measured by ESR, showed no significant differences. Thus microviscosity was the factor governing diffusion. Gelatin concentration in aqueous solutions without glycerol influenced microviscosity and hence diffusion. This is believed to be caused by dissolution of water-soluble fractions of the gelatin. Interstices in the gelatin matrix, though reduced in size when gelatin concentration is raised, are still too large to act as physical barriers to diffusing molecules. It is suggested that hydrated gelatin forms the matrix of glycerogelatin mixtures and that the interstitial fluid, through which migration occurs, consists almost entirely of glycerol and water. PMID- 2571696 TI - Pharmacokinetic modelling of the effect of activated charcoal on the intestinal secretion of theophylline, using the isolated vascularly perfused rat small intestine. AB - The effect of activated charcoal administration on the secretion of theophylline from the blood into the intestinal lumen has been examined by use of the rat isolated vascularly perfused small intestine. A closed two compartment model was used to analyse the vascular and luminal concentration-time curves obtained. An equation was derived to calculate the time-dependent intestinal clearance. From control experiments it was concluded that theophylline is secreted by a diffusional transport system through the intestinal wall. The intestinal clearance declined rapidly with time as a result of the concomitant increase in luminal theophylline concentration. After 120 min a steady state between the vascular and luminal perfusate was established. Administration of activated charcoal in the lumen had a profound effect on the kinetics of the drug. The vascular steady state concentration was depressed dramatically. The theophylline clearance remained nearly constant with time, because the blood to lumen concentration gradient was maximized. The maximal value for the intestinal theophylline clearance was estimated to be 0.88 mL min-1 and it equalled the value for the intestinal blood flow at the absorptive site. By use of the concept of absorptive site blood flow, the maximal effect of charcoal on systemic theophylline clearance could be adequately predicted for rats, dogs and man. Activated charcoal administration is only useful to enhance the systemic clearance of drugs or toxicants if that clearance is of the same order of magnitude as the absorptive site blood flow or lower. PMID- 2571697 TI - Involvement of different receptors in the central and peripheral effects of histamine on intestinal motility in the rat. AB - The effects of histamine on intestinal motility have been investigated in conscious rats, fed or fasted, using an electromyographic method. Histamine peripherally administered (10 mg kg-1) in 15 h fasted rats induced an inhibition followed by a period of irregular spiking activity disrupting the duodenojejunal migrating myoelectric complexes (MMC) and suppressed the postprandial motor spiking activity when administered 50 min after a meal. The selective agonist of the H1-receptors, 2-pyridylethylamine (2-PEA) induced an irregular spiking activity while dimaprit acting on H2-receptors, inhibited the MMC pattern. Effects of peripherally administered histamine were antagonized by previous administration of chlorpheniramine (0.5 mg kg-1 i.p.) and in a lesser extent by cimetidine (10 mg kg-1 i.p.). Histamine (1-10 micrograms) administered intracerebroventricularly (i.c.v.) in fasted rats increased the motor cycle frequency and immediately restored the MMC pattern when given to fed rats. Among the three agonists of the H1- H2- and H3-receptors (2-PEA, dimaprit and R-alpha methylhistamine, respectively) only R-alpha-methylhistamine (1-10 micrograms i.c.v.) was able to reproduce this effect. It is concluded that the effects of histamine on intestinal motility were centrally and peripherally mediated involving mainly H1-receptors at the peripheral level and H3-receptors at the CNS level. PMID- 2571698 TI - Alpha-blockade on blood pressure and on cardiac noradrenaline content in SHR and DOCA-salt rats. AB - The current study was designed to explore the effects of peripherally administered alpha-adrenoceptor inhibiting drugs (either prazosin or yohimbine) in spontaneously hypertensive (SHR) and DOCA-salt hypertensive rats before the development of established hypertension, as well as to characterize changes induced by prazosin and/or yohimbine in the cardiac noradrenaline (NA) content that might be responsible for the development of hypertension in these strains. The alpha-adrenoceptor blockade can prevent hypertension in DOCA and salt-treated animals in such a way that their blood pressure stabilizes at levels significantly lower than those observed in similarly treated normotensive controls. Significantly lower cardiac NA content was observed in DOCA-salt rats under basal and experimental conditions. The blood pressure of the treated rats and the heart NA content of the SHR were unaltered by treatment. Thus, administration of the alpha-adrenoceptor blocking agents, prazosin and/or yohimbine, throughout the developmental stage of SHR hypertension failed to alter either the progressive rise in blood pressure or in NA content. There may be differences between the cardiac adrenergic mechanisms responsible for the development of hypertension in each of these two models of hypertension. PMID- 2571699 TI - Interactions of apomorphine and a novel neuroleptic dibenzodioxazocine derivative, as evidenced by changes of somato-autonomic reflexes and spontaneous sympathetic activity in cats. AB - The apomorphine-antagonistic effects of EGYT-2509, a novel neuroleptic compound, has been studied by two different methods suitable for investigating the dopaminergic modulation of sympathetic output. (1) In cats lightly anaesthetized with urethane (600 mg kg-1 i.p.), blood pressure (BP) reflexes evoked by electrical stimulation of the sciatic nerve were inhibited by apomorphine (0.2 mg kg-1 i.v.) at low frequencies of stimuli (2-8 Hz), while the BP reflexes were facilitated by apomorphine at higher frequencies of stimulation; the evoked contractions of the nictitating membrane were depressed in the entire range of frequencies applied. EGYT-2509 (1.5 mg kg-1 i.v.) antagonized both the inhibition and facilitation of pressor reflexes induced by apomorphine. EGYT-2509, given alone, in doses exceeding 1.5 mg kg-1 either did not influence or inhibited the responses of nictitating membrane and of BP; the inhibition could be antagonized by haloperidol. The apomorphine-induced sustained hypotension was inhibited by EGYT-2509 (18.5 mg kg-1): after EGYT-2509, higher doses of apomorphine (0.7 vs 0.2 mg kg-1) were required for the effect. Sustained hypotension could be elicited by EGYT-2509, too; after apomorphine, smaller doses of EGYT-2509 (8.5 vs 18.5 mg kg-1) were enough to decrease BP. (2) In cats anaesthetized with chloralose and urethane (50 and 400 mg kg-1 i.p., respectively), apomorphine (0.2 mg kg-1) inhibited the spontaneous activity of the postganglionic renal sympathetic nerve.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571701 TI - High resolution electron microscopy study of sulphathiazole crystals. AB - A high resolution electron microscopy study was undertaken on samples of sulphathiazole obtained by recrystallization at 0 degrees C, 30 degrees C and 70 degrees C. Low magnification electron microscopy study of the crystals showed featureless morphology yet the resolved lattice images showed imperfections such as dislocations, lattice irregularities and regions of discontinuity. PMID- 2571700 TI - Repeated administration of antidepressants enhances agonist affinity for mesolimbic D2-receptors. AB - Studies have shown an increased responsiveness of the dopaminergic system after repeated administration of a variety of antidepressant drugs. In the present study, the effect of repeated administration (twice daily for 14 days) of imipramine and mianserin on the affinity of dopamine D2-receptors for quinpirole, a D2-agonist, and on the quinpirole-induced locomotor hyperactivity was examined in rats. Repeated doses of imipramine and mianserin increased the affinity of quinpirole for [3H]spiperone binding sites in membranes prepared from the limbic system but not the striatum. The locomotor hyperactivity induced by quinpirole was enhanced by chronic treatment with both antidepressants. The data indicate that the enhanced responsiveness of the dopaminergic system in rats, observed after chronic treatment with antidepressants, may result from an increased affinity of agonists at D2-receptors in the mesolimbic system. PMID- 2571702 TI - Pharmacokinetic study of tempo carboxylic acid, a nitroxyl MRI contrast media, in control and streptozocin diabetic rats. AB - The pharmacokinetics of tempo carboxylic acid (TCA), a nitroxyl contrast medium have been evaluated in control and streptozocin-diabetic rats. Previous magnetic resonance imaging (MRI) studies in diabetic rats showed prolongation of contrast visualization in the renal cavities after injection of TCA. Diabetes induced only slight alterations to the pharmacokinetic parameters. Rate constants and half lives were unchanged after four months of diabetes. A significant decrease of the apparent total body clearance and volume of distribution was observed while the area under the curve was increased. These alterations are not sufficient to explain MRI abnormalities which have to be elucidated. PMID- 2571703 TI - The protective mechanisms of paracetamol against ethanol-induced gastric mucosal damage in rats. AB - The protective mechanisms of paracetamol against ethanol-induced gastric mucosal damage have been examined. The antiulcer action of subcutaneously (s.c.)-injected paracetamol, 250 mg kg-1, was attenuated by either subdiaphragmatic vagotomy or s.c. injection of N-ethylmaleimide, 10 mg kg-1. This attenuation was not seen in rats given paracetamol by the oral route (p.o.). Indomethacin pretreatment, 5 mg kg-1, did not influence the lesion-preventing action of paracetamol given s.c. or p.o. The findings suggest that the antiulcer effect of s.c.-administered paracetamol results from an action involving the vagal nerve and tissue sulfhydryls, but not prostaglandins. On the other hand, the protective mechanism of paracetamol p.o. is independent of the vagal system or tissue sulfhydryls and prostaglandins. It seems that paracetamol given p.o. exerts its antiulcer effect by acting directly on the mucosal cell to strengthen mucosal integrity. PMID- 2571704 TI - The role of vagal adrenergic activity in the mechanism of gastric acid secretion after pylorus-ligation in the rat. AB - The role of the vagus nerve and adrenoceptor stimulation in acid secretion after pylorus-ligation in the rat has been examined. All drugs were administered intraperitoneally. Atropine (5 mg kg-1) depressed the H+ output (111 mumols +/- 33.8 vs 412.5 mumols +/- 62.2, mean +/- s.e.m., n = 10, P less than 0.001); cimetidine (40 mg kg-1) did not enhance this action, while vagotomy was more effective than atropine (32.7 mumols +/- 4.9, mean +/- s.e.m., n = 10, P less than 0.05). Atropine (10 mg kg-1) produced a similar depression to the 5 mg kg-1 dose. Cimetidine (100 mg kg-1) depressed the H+ output (248.5 mumols +/- 46.8, mean +/- s.e.m., n = 10, P less than 0.05). Propranolol (5-20 mg kg-1) had no significant effect on the H+ output but dose-dependent inhibition was produced by phenoxybenzamine or phentolamine; an inhibition similar to that achieved by vagotomy was seen with the 20 mg kg-1 dose. Both these drugs (5 or 10 mg kg-1) had no significant effect on the H+ output when given with atropine (5 mg kg-1) but the H+ output was significantly lower than that produced by either drug at the same dose given alone. Atropine (5 mg kg-1) with phenoxybenzamine or phentolamine (20 mg kg-1) produced H+ output not significantly different from that with vagotomy or either alpha-adrenoceptor given alone at 20 mg kg-1, but the result was significantly (P less than 0.05) lower than the H+ output with atropine (5 mg kg-1) alone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571705 TI - Effect of centrally administered prostaglandin D2 and some prostaglandin synthesis inhibitors on carrageenan-induced paw oedema in rats. AB - The putative modulatory role of central prostaglandins (PGs) on peripheral inflammation has been explored by investigating the effects of intracerebroventricularly (i.c.v.) administered PGD2, the major rodent brain PG, hydrocortisone, a phospholipase A2 inhibitor, and the cyclo-oxygenase inhibitors, paracetamol and mefenamic acid, on carrageenan-induced paw inflammation in rats. PGD2 produced a dose-related inflammation-augmenting effect, whereas hydrocortisone and the PG synthesis inhibitors, paracetamol and mefenamic acid, induced attenuation of the peripheral oedema. These findings confirm an earlier report from this laboratory which had indicated that central PGs may modulate peripheral inflammation and that conventional anti-inflammatory agents exert at least part of their effect by inhibiting central PG synthesis. PMID- 2571706 TI - Effects of metabolic inhibitors and incubation temperature on the saturable uptake of propranolol by isolated rat lung tissue. AB - The effects of several metabolic inhibitors (50 microM) on the initial uptake rate of propranolol (0.5 to 500 micrograms mL-1) by the minced lungs (0.4 g) isolated from 7-week-old rats has been investigated in oxygenated, pH 7.4 Krebs Ringer bicarbonate buffer solution (20 mL) containing 3% BSA at 37 degrees C for 5 min. The effect of the incubation temperature was also examined. Metabolism of propranolol was almost insignificant (i.e. less than 1.3% of the initial load). The overall initial uptake rate was considered to be a combination of apparent linear transport and saturable processes. For the control uptake rate, the linear transport rate constant was 1.26 +/- 0.16 g-1 mL-1 min-1, while Vmax and Km' of the capacity limited uptake process were estimated as 0.727 +/- 0.074 mg g-1 min 1 and 24.8 +/- 2.71 micrograms mL-1, respectively. No metabolic inhibitor tested had an effect on the linear transport rate of propranolol but 2,4-dinitrophenol and potassium cyanide inhibited saturable uptake rate (i.e. Vmax) of propranolol significantly (P less than 0.01) while ouabain, phloridine and iodoacetic acid did not do so significantly. Reduction of the incubation temperature to 15 degrees C decreased and at 25 degrees C tended to decrease, both linear transport and saturable uptake rates. PMID- 2571707 TI - Influence of capsaicin-sensitive fibres on experimentally-induced colitis in rats. AB - Systemic capsaicin pretreatment worsens trinitrobenzene sulphonic acid-induced colitis but has no effect on colitis induced by ethanol or acetic acid. The influence of capsaicin-sensitive fibres on experimentally-induced colitis seems to depend upon the type of ulcerogenic stimuli in relation to its chronic nature. PMID- 2571708 TI - Postnatal effects of maternal nicotine exposure on the striatal dopaminergic system in rats. AB - The biochemical effects of continuous prenatal exposure to nicotine (1.5 mg kg-1 day-1) have been examined in 14 day old male and female rat pups. The ability of striatal tissue slices from the nicotine-exposed pups to synthesize [3H]dopamine from [3H]tyrosine was enhanced. However, the ability of (+)-amphetamine to stimulate formation and release of [3H]dopamine from [3H]tyrosine was not affected. Furthermore, prenatal exposure to nicotine increased the rate of striatal dopamine turnover in the male, but not the female nicotine-exposed pups. PMID- 2571709 TI - The effect of Miglycol 812 oil on the oral absorption of propranolol in the rat. AB - This work has examined the effect of Miglyol 812 oil and its composite fatty acids on the oral absorption of propranolol with reference to its intravenous (i.v.) pharmacokinetics. Propranolol hydrochloride, spiked with 4-(3) H labelled compound, was administered i.v. or orally to male Wistar rats and blood concentrations of parent material determined by liquid scintillation counting after extraction into toluene. An i.v. dose-linearity study indicated dose independent pharmacokinetics for propranolol at 1-2 mg kg-1, with a mean Cls, Vss, MRTi.v. and t0.5 beta of 0.076 L min-1 kg-1, 4.74 L kg-1, 57.81 min and 47.10 min, respectively. At 5 mg kg-1, there was evidence of non-linearity with MRTi.v. increased by about 250%, Vss by 170% and t0.5 beta by 230% compared with the lower doses. After oral administration of propranolol (10 mg kg-1) in aqueous solution, with or without Tween 80 (6%), the mean absorption time (MAT) and terminal half-life were approximately 55 min and 86 min, respectively. The MAT for propranolol administered in a 50% octanoic and lauric acid (1:1 by weight) oil-in-water emulsion, stabilized with 6% Tween 80 (129-90 min), was significantly longer compared with that for a 50% Miglyol 812 oil-in-water emulsion containing the same surfactant (16.55 min). The terminal half-life of propranolol administered in the fatty acid formulation (128.96 min), unlike that for the Miglyol emulsion (54-37 min), was significantly longer compared with that observed after i.v. administration (t0.5 beta = 47 min).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571710 TI - The effect of varying extents of Fluosol-DA or normal saline haemodilution on the dose dependent kinetics of phenytoin in the rat. AB - Phenytoin kinetics were determined in rats in which the blood was moderately haemodiluted with 20 or 40 mL kg-1 of Fluosol-DA or normal saline. Rats received one of three intravenous phenytoin doses (10, 40, 50 mg kg-1) 0.5, 24, 48, or 72 h after haemodilution and were compared with non-exchanged controls. Haemodilution with either 20 or 40 mL kg-1 of Fluosol or saline had no influence on the dose-dependent kinetics of phenytoin. Haemodilution with 40 mL kg-1 of Fluosol decreased the half-life of phenytoin's major metabolite, HPPH, after a 50 mg kg-1 dose. Neither Fluosol nor saline haemodilution affected the normal delay in biliary cycling of HPPH. PMID- 2571711 TI - Second generation model for prednisolone pharmacodynamics in the rat. AB - An improved model describing receptor/gene-mediated pharmacodynamics of prednisolone is presented which consists of seven differential equations. Data for plasma prednisolone concentrations, free hepatic glucocorticoid receptors, and hepatic tyrosine aminotransferase activity (TAT) following low (5 mg/kg) and high (50 mg/kg) doses of prednisolone are used to quantitate the kinetics and dynamics of this synthetic steroid in the rat. In contrast to the earlier model, the newer model provides for a coupling and simultaneous fitting of receptor and TAT data and is able to describe the recycling of receptors between cytosol and nucleus and the return of cytosolic receptors to baseline following glucocorticoid elimination. A numerical technique to determine the efficiency of TAT induction based on area under the curve calculations is presented, which supports the hypothesis that nonlinear dose-response effects are due to dose and time-dependent receptor depletion in the cytosol. Simulations are presented to examine the major determinants of corticosteroid effects and to compare the effects of single- and multiple-dose regimens in maximizing drug effects. PMID- 2571712 TI - CI-943, a potential antipsychotic agent. I. Preclinical behavioral effects. AB - CI-943 (8-ethyl-7,8-dihydro-1,3,5-trimethyl-1H-imidazo[1,2-c] pyrazolo[3,4-e] pyrimidine) is a novel agent that is chemically unrelated to available antipsychotics and is not a dopamine receptor antagonist. Like available antipsychotics, CI-943 reduces spontaneous locomotion in mice and rats and inhibits compulsive cage climbing induced by apomorphine in mice at doses that do not produce ataxia. However, CI-943 enhances rather than inhibits the locomotor stimulant effects of d-amphetamine in mice and rats. Unlike dopamine antagonists, CI-943 does not affect stereotypy caused by apomorphine or amphetamine in rats. CI-943 displays an antipsychotic-like profile in conditioned avoidance tests, inhibiting one-way avoidance in rats at doses that do not impair escape and inhibiting continuous avoidance in rats and squirrel monkeys at doses that do not impair shock termination responding. Although high doses of CI-943 produce dystonic movements in haloperidol-sensitized monkeys, CI-943 differs from dopamine antagonists that produce extrapyramidal dysfunction in humans in that doses of CI-943 that are sufficient to inhibit avoidance responding in monkeys do not produce extrapyramidal dysfunction. Unlike dopamine antagonists that produce tardive dyskinesia, CI-943 administered repeatedly at high doses does not produce behavioral supersensitivity to dopamine agonists in rats. These results demonstrate that CI-943 resembles available antipsychotics in some preclinical behavioral tests commonly used to predict antipsychotic efficacy but differs from dopamine antagonists in tests predictive of dopamine receptor antagonism and antipsychotic-induced neurological dysfunction. PMID- 2571713 TI - CI-943, a potential antipsychotic agent. II. Neurochemical effects. AB - The effects of CI-943 (a novel 8-ethyl-7,8-dihydro-1,3,5-trimethyl-1H-imidazo[1,2 c]pyrazolo[3,4- e]pyrimidine compound exhibiting a favorable antipsychotic profile in animal tests) on neurochemical parameters related to biogenic amine neurons have been studied in rat brain. CI-943 (1-40 mg/kg p.o. and 20 mg/kg i.p.) accelerated the turnover of dopamine (DA) in rat brain as demonstrated by the enhancement of levels of the DA metabolites homovanillic acid, 3,4 dihydroxyphenylacetic acid or 3-methoxytyramine and by the enhancement rate of DA synthesis in either striatum or mesolimbic regions. These increases in DA turnover induced by CI-943 are not due to DA receptor blockade as CI-943, unlike known antipsychotics, did not exhibit affinity for DA receptors either in vitro or in vivo and did not affect rat serum basal prolactin levels. Amfonelic acid enhanced the action of haloperidol in increasing striatal homovanillic acid with no effect on that of CI-943 and clozapine, suggesting that CI-943, like clozapine, would be predicted to have a low risk of extrapyramidal side effects as compared to haloperidol. Chronic administration of CI-943 (40 mg/kg i.p.) to rats for 28 days did not affect the affinity or number of striatal DA receptors; in comparison haloperidol (0.5 mg/kg i.p.) caused an increase in number of DA receptors with no change in affinity. Measures of serotonergic function were increased; noradrenergic function was not affected by CI-943, nor did it exhibit affinity for a number of central nervous system receptors in vitro. The molecular mechanism by which CI-943 increases brain DA turnover is not known at this time but appears to be unique in comparison to known antipsychotic agents. PMID- 2571714 TI - CI-943, a potential antipsychotic agent. III. Evaluation of effects on dopamine neuronal activity. AB - CI-943 (8-ethyl-7,8-dihydro-1,3,5-trimethyl-1H-imidazo[1,2- c]pyrazolo[3,4 e]pyrimidine) has been identified as a novel potential antipsychotic agent that does not bind to dopamine (DA) receptors. In the present studies, the effects of acute and chronic administration of CI-943 on the electrophysiological activity of A9 and A10 DA neurons were assessed and compared to the effects of DA antagonist antipsychotic drugs. Acute administration of CI-943 did not increase the base-line firing rate (10-20 mg/kg i.p.), did not increase the number of spontaneously active DA neurons (40 mg/kg p.o.) and did not antagonize the effects of apomorphine or amphetamine on A9 or A10 DA neurons (20-40 mg/kg i.p.). A high dose of CI-943 (40 mg/kg i.p.) decreased the firing rate of A9 and A10 DA neurons, an effect that was not antagonized by haloperidol. In contrast, haloperidol increased the base-line firing rate (0.5 mg/kg i.p.), increased the number of spontaneously active DA neurons (0.5 mg/kg p.o.) and antagonized the effects of apomorphine and d-amphetamine on A9 and A10 DA neurons (0.1 mg/kg i.p.). Repeated (21 days) administration of CI-943 (36 mg/kg/day p.o.) did not alter the number of spontaneously active A9 DA neurons but increased the number of active A10 DA neurons. In contrast, repeated administration of haloperidol (0.4 mg/kg/day p.o.) decreased the number of spontaneously active DA neurons in A9 and A10, whereas repeated administration of clozapine (19 mg/kg/day p.o.) decreased the number of active A10 DA neurons but increased the number of active A9 DA neurons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571715 TI - Inhibition of rat brain glutamate receptors by philanthotoxin. AB - The actions of philanthotoxin (PhTX) were studied on the function of glutamate receptors expressed in Xenopus oocytes injected with rat brain mRNA and on binding of radioligands to rat brain glutamate receptors. PhTX reversibly inhibited the oocyte responses to quisqualate, N-methyl-D-aspartate (NMDA) and kainate in a dose-dependent manner. The NMDA receptor was the most sensitive to PhTX action (10-fold more than the kainate receptor) and the least sensitive was the smooth current component of the quisqualate response. Recovery from PhTX block differed among the three amino acids. NMDA responses recovered completely within a few minutes whereas responses to kainate and quisqualate recovered more slowly. PhTX had no effect on equilibrium binding of [3H]glutamate to rat brain cortical membranes studied in buffer treated to eliminate microorganisms. Based on the drug specificity of this [3H]glutamate binding, it is suggested to be mostly to the NMDA receptor. Low concentrations of PhTX (1-10 microM) potentiated binding of [3H] MK-801, a specific noncompetitive inhibitor of the NMDA receptor. However, higher PhTX concentrations inhibited this binding with an IC50 of 20 microM, similar to its inhibition of the oocyte-expressed NMDA receptor. Inhibition of [3H]MK-801 binding by PhTX was noncompetitive. It is suggested that PhTX, like the more potent MK-801, binds to an allosteric site on the NMDA receptor and inhibits its function but its binding site is not identical with the MK-801 binding site. PMID- 2571716 TI - Pharmacological distinctions between functional nicotinic acetylcholine receptors on the PC12 rat pheochromocytoma and the TE671 human medulloblastoma. AB - Some properties of functional nicotinic acetylcholine receptors (nAcChoR) expressed by the PC12 rat pheochromocytoma or the TE671 human medulloblastoma were studied by the use of an isotopic rubidium ion efflux assay. The assay involves active uptake of 86Rb+ via a ouabain-sensitive mechanism to load cells with isotopic tracer and the subsequent release of ion from cells through agonist activated opening of nAcChoR-coupled ion channels. For either cell line the rate of receptor-mediated ion efflux is time-dependent and falls as duration of exposure to agonist increases. However, dose-response curves for agonist activation of receptor function (or for antagonist blockade of agonist activation) are temporally invariant. Dose-response curves have characteristic shapes for a given agonist, and the relative and absolute potencies of agonists differ between TE671 and PC12 cells. Most notably, nicotine and cytisine are more potent activators of receptor function on the PC12 cell line whereas TE671 cell nAcChoR are more sensitive to activation by isoarecolone and suberyldicholine. PC12 cell nAcChoR are more sensitive to blockade by the classic "ganglionic" blockers, mecamylamine and hexamethonium, and by the molluscan substance, neosurugatoxin, than are nAcChoR on TE671 cells. The results, illustrating differences in the pharmacological profiles of drugs acting as functional nAcChoR on TE671 and PC12 cells, suggest that structural differences exist in nAcChoR active site(s) and are consistent with the notion that functional nAcChoR are a heterogeneous family of macromolecules. PMID- 2571717 TI - Classification of typical and atypical antipsychotic drugs on the basis of dopamine D-1, D-2 and serotonin2 pKi values. AB - The pKi values of 13 reference typical and 7 reference atypical antipsychotic drugs (APDs) for rat striatal dopamine D-1 and D-2 receptor binding sites and cortical serotonin (5-HT2) receptor binding sites were determined. The atypical antipsychotics had significantly lower pKi values for the D-2 but not 5-HT2 binding sites. There was a trend for a lower pKi value for the D-1 binding site for the atypical APD. The 5-HT2 and D-1 pKi values were correlated for the typical APD whereas the 5-HT2 and D-2 pKi values were correlated for the atypical APD. A stepwise discriminant function analysis to determine the independent contribution of each pKi value for a given binding site to the classification as a typical or atypical APD entered the D-2 pKi value first, followed by the 5-HT2 pKi value. The D-1 pKi value was not entered. A discriminant function analysis correctly classified 19 of 20 of these compounds plus 14 of 17 additional test compounds as typical or atypical APD for an overall correct classification rate of 89.2%. The major contributors to the discriminant function were the D-2 and 5 HT2 pKi values. A cluster analysis based only on the 5-HT2/D2 ratio grouped 15 of 17 atypical + one typical APD in one cluster and 19 of 20 typical + two atypical APDs in a second cluster, for an overall correct classification rate of 91.9%. When the stepwise discriminant function was repeated for all 37 compounds, only the D-2 and 5-HT2 pKi values were entered into the discriminant function.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571719 TI - Attenuation by dopamine uptake blockers of the inhibitory effects of 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine and some of its analogs on NADH-linked metabolism in mouse neostriatal slices. AB - In previous studies it was shown that the pyridinium species formed by the monoamine-oxidase-catalyzed conversion of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) and 1-methyl-4-(2'methylphenyl)-1,2,3,6 tetrahydropyridine (2'Me-MPTP) inhibited mitochondrial electron transport at Complex I. In addition, these substances, when incubated with mouse neostriatal slices, caused an increased lactate accumulation. However, it was not clear whether this inhibition could occur within dopamine nerve terminals. In the present study we investigated if dopamine uptake blockers, shown previously to protect against tetrahydropyridine-induced dopaminergic neurotoxicity, would attenuate MPTP- and 2'Me-MPTP-promoted lactate formation in neostriatal slices. 2'Me-MPTP-induced lactate accumulation in neostriatal slices from mice or rats with a lesion of the nigrostriatal pathway was also studied. The dopamine uptake blocker, Win 35,428 [8-azabicyclo[3.2.1]octane-2-carboxylic acid, 3-(4 fluorophenyl)-8-methyl-, methylester [1R-(exo, exol)]), attenuated the increased lactate formation caused by MPTP and 2'Me-MPTP. At low concentrations of 2'Me MPTP (5 microM), the increased lactate production was inhibited completely by Win 35,428. These latter data suggest that at low concentrations of 2'Me-MPTP, the increased lactate accumulation was associated primarily with dopaminergic nerve terminals. Two other dopamine uptake inhibitors, McN 5908 [trans-4-(1,2,3,5,6,10b hexahydropyrrolo[2,1-alpha]isoquinolin+ ++-6-yl) benzenamine hydrobromide methanolate (4:4:1)] and mazindol [5-(4-chlorophenyl)-2,5-dihydro-3H-imidazo- [2,1-alpha]isoindol-5-ol], also attenuated tetrahydropyridine-induced lactate formation. At concentrations selective for their respective uptake systems, norepinephrine- and serotonin-uptake inhibitors did not attenuate 2'Me-MPTP induced lactate accumulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571718 TI - Effects of alpha adrenoceptor and dopamine receptor agonists and antagonists on ganglionic transmission. AB - Effects of alpha adrenoceptor and dopamine (DA) receptor agonists and antagonists on ganglionic transmission were studied in pentobarbital-anesthetized, open-chest dogs. Changes in tachycardia produced by preganglionic cardiac nerve stimulation were monitored as a measure of ganglionic transmission. Several agonists, injected i.a. into the blood supply of the stellate ganglion, inhibited ganglion transmission. This effect was localized to the ganglion as none of the agonists, in the highest doses studied, inhibited tachycardia produced by postganglionic cardiac nerve stimulation. The potency order of the agonists was UK 14,304 (alpha 2 adrenoceptor agonist) greater than norepinephrine (NE) greater than dipropyl DA (DA2 dopamine receptor agonist) greater than DA greater than or equal to phenylephrine (alpha-1 adrenoceptor agonist) greater than fenoldopam (DA1 dopamine receptor agonist). UK 14,304 was over 200 times more potent than fenoldopam, being active in nanomole doses, whereas NE was more potent than DA. Rauwolscine, an alpha-2 adrenoceptor antagonist, antagonized the inhibitory effects of NE and DA and was the only antagonist, given i.a. or i.v., that facilitated ganglionic transmission; frequency-response curves of tachycardia induced by preganglionic nerve stimulation were dose-dependently augmented. SCH 23390, in double the full DA1 antagonist dose, had no effect on frequency response curves or on NE- or DA-induced ganglionic inhibition. Domperidone antagonized the effect of DA but had no effect on ganglionic inhibition produced by NE or on frequency-response curves of tachycardia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571721 TI - Identification and characterization of presynaptic and postsynaptic beta adrenoreceptors in the longitudinal smooth muscle/myenteric plexus of dog ileum. AB - The subcellular distribution of [125I]cyanopindolol (125I-CYP) binding in membrane fractions derived from the longitudinal smooth muscle/myenteric plexus (LSM/MP) of dog ileum followed a bimodal pattern with selective enrichments in the purified synaptosomal (S2) fraction and the purified smooth muscle plasma membrane (M2) fraction. The half-maximal saturation of binding sites in both fractions occurred at 10 pM [125I]CYP with maximal binding capacities, in femtomoles per milligram of protein, 70 +/- 10 (S2 fraction) and 110 +/- 25 (M2 fraction). Approximately 80% of binding sites in either membrane fraction exhibited a high affinity for beta-2 selective antagonist ICI 118,551 (Ki = 1.4 +/- 0.4 nM in S2 and 1.8 +/- 0.6 nM in M2; mean +/- S.D. n = 3), whereas remaining 20% of binding sites had a low affinity for this agent (Ki = 2.6 +/- 1.5 microM in S2 and 0.55 +/- 0.2 microM in M2). The beta-1-selective antagonist ICI 89,406 had a high affinity for approximately 20% of binding sites (Ki = 12 +/ 7 nM in S2 and 4 +/- 2 nM in M2) and a low affinity for approximately 80% of binding sites (Ki = 0.34 +/- 0.16 microM in S2 and 0.42 +/- 0.24 microM in M2). The displacement potencies of beta-adrenergic agonists followed the order (-) isoproterenol greater than salbutamol (-)-epinephrine greater than (-) norepinephrine in each membrane preparation. The inclusion of 0.1 mM 5' guanylylimidodiphosphate into assay medium resulted in a decreased affinity of receptors for agonists and increased values of Hill coefficients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571720 TI - Pharmacological characterization of alpha adrenoceptors involved in the antinociceptive and cardiovascular effects of intrathecally administered clonidine. AB - The effects on nociception, blood pressure and heart rate of clonidine administered intrathecally to the lumbar level were determined in conscious rats and in rats anesthetized lightly with pentobarbital. In anesthetized rats, intrathecal (i.t.) clonidine (3.2-32.0 micrograms) inhibited the nociceptive tail flick reflex and had biphasic effects on blood pressure; lesser doses (1.0-10.0 micrograms) produced depressor effects, whereas a greater dose (32.0 micrograms) produced a marked pressor response. Clonidine also produced biphasic effects on blood pressure in conscious rats, with the dose-response function shifted upward and to the left of that observed in anesthetized rats. The depressor and antinociceptive effects of 3.2 micrograms of clonidine were antagonized by pretreatment with yohimbine (30.0 micrograms i.t.) but not by prazosin (30.0 micrograms i.t.) or by yohimbine (0.1 mg/kg i.v.). Thus, these effects of clonidine are mediated by spinal alpha-2 adrenoceptors. The pressor response to 32.0 micrograms of clonidine (i.t., lumbar) was accompanied by marked bradycardia, and similar cardiovascular effects were observed when this dose of clonidine was administered either i.v. or to the cervical level of the spinal cord. The pressor response to 32.0 micrograms of clonidine (i.t., lumbar) was not reduced significantly by i.t. pretreatment with yohimbine (30.0 micrograms) or prazosin (30.0 micrograms), but was diminished significantly by i.v. pretreatment with yohimbine (1.0 mg/kg), prazosin (0.1 mg/kg) or phentolamine (2.0 mg/kg). Neither chlorisondamine (2.5 mg/kg i.v.) or the V1-vasopressin receptor antagonist [1-(beta-mercapto-beta,beta-cyclopentamethylene propionic acid), 2-(o methyl)tyrosine]Arg8-vasopressin (10.0 micrograms/kg i.v.) reduced the clonidine produced pressor response. After i.t. injection of 32.0 micrograms of [3H]clonidine, peak levels of radioactivity in the blood were observed at 2 min and corresponded to a blood concentration of 38.8 ng/ml. Injection of an i.v. bolus dose (2.5 micrograms/kg) sufficient to produce these blood levels resulted in a transient pressor response. These results suggest that after i.t. administration of greater doses of clonidine, sufficient amounts of the drug are rapidly redistributed systemically to produce pressor effects by stimulation of vascular alpha adrenoceptors. PMID- 2571722 TI - Kappa-opioid receptor-mediated antinociception in the rat. I. Comparative actions of mu- and kappa-opioids against noxious thermal, pressure and electrical stimuli. AB - This study examined the comparative efficacy of mu- and kappa-opioids in modulating the tail-withdrawal response to matched intensity heat and pressure stimuli and the vocalization response to electrical stimulation in the rat. All drugs were given s.c. The mu-agonists, morphine and fentanyl, were equipotent against heat and pressure at all intensities. The kappa-agonists, U50,488H and U69,593 also acted against pressure in an intensity-independent fashion. However, their action against heat was intensity-dependent being greatest against "low" and least against "high" intensity. At "moderate" intensities, morphine, fentanyl, U69,593, U50,488H and 2 other kappa-agonists (bremazocine and tifluadom) were equipotent against heat and pressure. The action of tifluadom was stereospecific. Naltrexone (a preferential mu-antagonist) was 10 times more potent in blocking the actions of morphine and fentanyl (as compared to U69,593 and U50,488H) against both heat and pressure. The preferential kappa-antagonists, nor-binaltorphimine and MR 2266, were 5- and 2-fold, respectively, more potent against the kappa-ligands for both heat and pressure. The antagonism of MR 2266 was expressed stereospecifically. Fentanyl and morphine induced a powerful antinociception against electrical stimulation whereas U69,593, U50,488H, bremazocine and tifluadom were completely inactive. Thus, kappa-opioid receptors can mediate antinociception against noxious thermal stimuli. However, in contrast to mu-receptors, this antinociception against heat is intensity-dependent. Kappa agonists, in distinction to mu-agonists, are inactive against electrical stimulation. PMID- 2571723 TI - Kappa-opioid receptor-mediated antinociception in the rat. II. Supraspinal in addition to spinal sites of action. AB - This study examines whether there is a supraspinal, in addition to spinal, component to the antinociceptive actions against heat and pressure stimuli of kappa-opioid receptor agonists (U-69,593, U50,488H, bremazocine and tifluadom) as compared to mu-opioid receptor agonists (Tyr-D-Ala-Gly-NMe-Gly-ol, fentanyl and morphine) in the rat. The antinociception induced by kappa- and mu-opioids (applied s.c.) was unaffected by systemic quaternary naltrexone (50 mg/kg) revealing that it is mediated in the central nervous system. All kappa- and mu opioids produced dose-dependent antinociception upon intrathecal application, in each case reversible by naloxone (5 mg/kg s.c.). However, intrathecal application of naloxone could only partially (by ca. 50%) antagonize the antinociception evoked by systemically applied U50,488H and morphine: this suggests sites of action in brain in addition to spinal cord for both mu- and kappa-opioids. Intraventricular application of mu-agonists produced maximal, dose-dependent antinociception. All kappa-agonists were also active in producing dose-dependent antinociception although curves were shallow and maximal antinociception could not be attained. The action of tifluadom was shown to be stereospecific. Naltrexone was 10-fold more potent in blocking morphine as compared to U50,488H whereas nor-binaltorphimine, a preferential kappa-antagonist, was 6-fold more potent against U50,488H than morphine. Indeed, whereas a dose of 0.2 mg/kg of naltrexone reversed mu-agonist actions, this dose was inactive against all kappa agonists: the actions of these could be antagonized only by 2.0 mg/kg. These data indicate that in addition to kappa-receptors in the spinal cord, kappa-receptors in the brain can mediate antinociception against noxious heat and pressure. PMID- 2571724 TI - Absorptive-mediated endocytosis of a dynorphin-like analgesic peptide, E-2078 into the blood-brain barrier. AB - The binding and internalization of a novel analog of dynorphin-like analgesic basic peptide, [125I]E-2078 (CH3-[125I] Tyr-Gly-Gly-Phe-Leu-Arg-CH3Arg-D-Leu NHC2H5), by isolated bovine brain capillaries were investigated. High-performance liquid chromatographic analysis showed that no significant metabolism of [125I] E 2078 occurred during incubation with brain capillaries for 30 min at 37 degrees C. The binding of [125I]E-2078 to brain capillaries increased with time and the steady-state cell-to-medium concentration ratio was 58.5 +/- 2.6 microliters/mg of protein. Approximately one-fourth of the [125I]E-2078 binding was resistant to acid wash, and showed significant dependence on temperature and medium osmolarity. The acid sensitive binding of [125I]E-2078, which presumably represents surface binding, was saturable and the Scatchard plot gave a maximal binding capacity Bmax = 147 +/- 29 pmol/mg of protein, and a half-saturation constant (KD) = 4.62 +/- 0.59 microM. Pretreatment of brain capillaries with phenylarsine oxide, an endocytosis inhibitor, completely suppressed the acid resistant binding of [125I]E-2078, but did not influence the surface binding of [125I]E-2078. The acid resistant binding of [125I] E-2078 was inhibited by poly-L lysine and protamine, but not inhibited by insulin, transferrin, dynorphin (1-8), beta-neoendorphin, naloxone or poly-L-glutamate. Moreover, in vivo brain extraction of [125I]E-2078 in rats was 368 +/- 55% higher than that of [3H] sucrose and was significantly inhibited by 1 mM of unlabeled E-2078. These results demonstrate that E-2078 is internalized by brain capillaries via absorptive-mediated endocytosis, which is a polycation-sensitive pathway. PMID- 2571725 TI - Ammonium ions cause relaxation of isolated canine arteries. AB - Experiments were designed to determine the mechanism of action underlying relaxation of vascular smooth muscle induced by ammonium ions. In particular, the possibility that these ions might be an endothelium-derived relaxing factor was examined. Rings of large canine femoral, mesenteric and coronary arteries and of small arteries from the gracilis muscle were suspended in organ chambers for the recording of isometric force. Membrane potential was recorded with intracellular microelectrodes in smooth muscle cells from the mesenteric artery. Ammonium ions induced relaxation which were independent of the presence of the endothelium. The relaxations were not prevented by adrenergic, serotonergic, muscarinic and histaminic blockers, by scavengers of oxygen-derived radicals or by inhibitors of soluble guanylate cyclase. The relaxations were prevented by a decrease in extracellular calcium concentration and by inhibition of the Na+/K+ pump. The results are compatible with the hypothesis that the relaxation induced by ammonium ions is related to changes in intracellular pH and, at high concentration of these ions, possibly to activation of the Na+/K+ pump. Ammonium ions are neither the endothelium-derived relaxing factor which activates guanylate cyclase nor the factor that induces endothelium-derived hyperpolarization. Inasmuch as relatively low concentrations of the ion induce relaxation of small arteries of skeletal muscle, they could contribute to exercise hyperemia. PMID- 2571726 TI - Interpersonal behavior and cardiovascular reactivity in pharmacologically-treated hypertensives. AB - This study examined the interpersonal behavior and concomitant cardiovascular reactivity (CVR) of hypertensive patients whose resting blood pressure was controlled by antihypertensive medication. Thirty hypertensive and 30 normotensive subjects matched for age, sex and occupational status were recruited from an industrial setting. The groups were compared on measures of interpersonal behavior, blood pressure and heart rate while they engaged in both role-played (RP) and naturalistic interactions (NI) requiring positive and negative assertion. Interpersonal behavior of the groups was generally similar, with two exceptions: hypertensives made fewer requests for new behavior in the negative RP and verbalized more praise statements in the positive NI. There were no differences between the groups on measures of CVR during interpersonal interactions. Overall effectiveness during scenes requiring negative assertion was associated with increased cardiovascular reactivity, especially for hypertensives. The interpersonal behavior and cardiovascular responses of patients taking beta-blocker medication did not differ from those taking diuretics. These findings are discussed with regard to methodological considerations pertinent to the assessment of interpersonal behavior and the issue of heterogeneity among hypertensives. PMID- 2571727 TI - 5-Piperazinylalkyl-2(3H)-oxazolones with neuroleptic activity. AB - A series of new 4-aryl-5-[omega-(4-aryl-1-piperazinyl)alkyl]-2(3H)- oxazolones was synthesized and tested for neuroleptic activity in mice and rats. Several compounds exhibited interesting neuroleptic activity with very low liability to the extrapyramidal side effects. In particular the activity of 4-(4-fluorophenyl) 5-[2-[4-(3,5-dichlorophenyl)-1- piperazinyl]ethyl]-2(3H)-oxazolone (14) was greater than that of butropipazone and fluanisone, while of the same order of that of chlorpromazine; however, the product showed a longer lasting activity and minor ability to produce catalepsy as compared with the reference drugs. PMID- 2571729 TI - Resolved 6,7,8,9-tetrahydro-N,N-dimethyl-3H-benz[e]indol-8-amine: central dopamine and serotonin receptor stimulating properties. AB - The enantiomers of 6,7,8,9-tetrahydro-N,N-dimethyl-3H-benz[e]indol-8- amine (1a) were prepared and tested for their actions on central dopamine and serotonin (5 HT) receptors. The dopaminergic effects were shown to reside in the (1)-R enantiomer. It was shown that compound 1a and its (+)-R enantiomer possess potent central 5-HT1A receptor stimulating properties. PMID- 2571728 TI - Sterically hindered 5,11-dicarbo analogues of clozapine as potential chiral antipsychotic agents. AB - Sterically hindered 5,11-dicarbo analogues of clozapine were prepared as potential chiral antipsychotic agents, with the possibility that for a particular analogue the antipsychotic activity of clozapine may reside in one enantiomer of the analogue whereas other unwanted biological effects of clozapine may be caused by the other enantiomer. Variable-temperature proton nuclear magnetic resonance studies showed that although 5-methylene-10-(4-methylpiperazino)-5H dibenzo[a,d]cycloheptene exists at room temperature as configurational enantiomers, the activation energy for thermal racemization is 19 kcal mol-1 at 105 degrees C, and it is doubtful that the enantiomers of this analogue can be isolated under usual laboratory conditions. The (Z)-5-ethylidene and 5 isopropylidene analogues have activation energies greater than 23 kcal mol-1 at 160 degrees C, and thus there is a possibility that the analogues can be obtained as their respective enantiomers. 5-Methyl-10-(4-methylpiperazino)-5H dibenzo[a,d]cycloheptene incorporates a chiral center which is not thermally racemized, but it exists at room temperature as two diasteromers with an activation energy for inversion of the 5H-dibenzo[a,d]cycloheptene ring of 21 kcal mol-1. When the 5,11-dicarbo analogues were tested in vitro for biological activity and their activities were compared to that of clozapine, the affinities for muscarinic and dopamine D-1 and D-2 sites were reduced but were still substantial. Thus the respective biological activities of the racemates indicate that the biological activities of the thermally stable enantiomers may be of importance in finding a clozapine derivative with fewer side effects than those shown by clozapine itself. Because of the susceptibility of the enamines to acid catalyzed hydrolysis, resolution into respective enantiomers is not anticipated. PMID- 2571730 TI - The xanthene-9-spiro-4'-piperidine nucleus as a probe for opiate activity. AB - A series of novel 1'-methylxanthene-9-spiro-4'-piperidines has been prepared in the search for opiate analgesics with improved pharmacological properties. It has been found that introduction of a hydroxyl group into the 4-position of the xanthenespiropiperidine nucleus produces a potent mu-opiate agonist. The structure-activity relationship of the series has been explored by use of isosteric replacements of the phenolic hydroxyl group. Moreover, the effect of altering the conformation of the piperidine ring has been studied. It was interesting to note that, in compounds lacking the phenolic hydroxyl group, opiate activity could be produced by introduction of the (phenylamino)ethyl group instead of methyl at the 1'-position. PMID- 2571731 TI - Synthesis and pharmacological evaluation of CNS activities of [1,2,3]triazolo[4,5 b][1,5]-, imidazolo[4,5-b][1,5]-, and pyrido[2,3-b][1,5]benzodiazepines. 10 Piperazinyl-4H-1,2,3-triazolo[4,5-b][1,5]benzodiazepines with neuroleptic activity. AB - The synthesis of [1,2,3]triazolo[4,5-b][1,5]-, imidazolo[4,5-b][1,5]-, and pyrido[2,3-b][1,5]benzodiazepines is described. The antidopaminergic and anticholinergic activities of the compounds have been examined by the respective in vitro [3H]spiperone and [3H]QNB receptor binding assay. The neuroleptic potential has been further evaluated in terms of their ability to produce hypothermia and catalepsy in mice and a conditioned avoidance response in rats. Only compounds from the triazolobenzodiazepine series show antipsychotic potential. The lack of activity in the imidazolo- and pyridobenzodiazepine series indicates that the basicity of the heteroarene moiety may be determinant for activity. PMID- 2571732 TI - Substituted 5-amino-4,5,6,7-tetrahydroindazoles as partial ergoline structures with dopaminergic activity. AB - Two series of tetrahydroindazoles were synthesized and evaluated for dopaminergic activity. A number of these partial ergoline analogues possess substituents that could mimic the C-8 substituent of the dopaminergic ergolines. Of the unsymmetrically substituted amine series 7a-k, the (monopropylamino)tetrahydroindazole 7b was most interesting as it was found to selectively activate the dopamine (DA) autoreceptor at a dose of 5 mg/kg in rats. The disubstituted amines 7g-k had significant DA postsynaptic activity as measured by increases of serum corticosterone levels in rats. The 6-substituted-5 aminotetrahydroindazoles 10a-d were found to possess only marginal dopaminergic activity. PMID- 2571733 TI - Yearly change in antibody level to Dirofilaria immitis (Spirurida: Filariidae) among humans exposed naturally to infected mosquitoes. AB - The IgG antibody level to Dirofilaria immitis (Leidy) increased significantly in 10.0% of 558 paired sera collected yearly from inhabitants in Miki City, Japan, indicating that they were exposed to infected mosquito bites. An additional 15.2% of paired sera showed decreases in IgG level. The binominal distribution of ELISA antibody levels among paired sera suggested an equilibrium between continued infection and antibody decreases after infection. PMID- 2571734 TI - Lusitropic effect and modifications of contraction-relaxation coupling induced by alpha-adrenergic stimulation in rat left ventricular papillary muscle. AB - Phenylephrine (PE) and metaraminol (MR) were studied alone at 2 x 10(-5) M and at 4 x 10(-5) M respectively. These drugs were also used both in the presence of either propranolol (PR) at 4 x 10(-7) M (PE/PR and MR/PR groups) or prazosin (PZ) at 2 x 10(-7) M (PE/PZ and MR/PZ groups). Specific alpha-adrenergic stimulation (AS) was induced in the PE/PR and MR/PR groups. These AS were evaluated in isotonic and isometric conditions on rat left ventricular papillary muscle. Peak shortening velocity (Vcmax) and peak lengthening velocity (Vrmax) were calculated from the twitch with preload only. Positive (+dF/dtmax) and negative (-dF/dtmax) peak derivative forces were calculated from the isometric twitch. Two coefficients R1 and R2 were used to measure the coupling between contraction and relaxation at low and heavy load, respectively: R1 = Vcmax/Vrmax and R2 = (+dF/dtmax)/(-dF/dtmax). In all groups, there was a significant positive inotropic effect. As compared to control values before AS, R1 significantly decreased in all groups, (PE/PR: -15%; MR/PR: -18%; PE/PZ: -8%; MR/PZ: -23%; PE: 19%; MR: -32%). On the other hand, R2 significantly decreased only in three groups (PE/PZ: -5.4%; MR/PZ: -16.5%; MR: -12.0%) whereas it did not significantly change in the three other groups (PE/PR; MR/PR; PE). In all groups, and at low load, Vrmax increased more than Vcmax (positive relaxant effect i.e. R1 decreased). At heavy load, despite the positive inotropic effect, there was no significant relaxant effect after predominent alpha-AS. These results indicate that alpha-AS modified the coupling between contraction and relaxation differently, depending on the level of load. PMID- 2571735 TI - Collateral resistance to verapamil in multidrug-resistant mouse tumor cells. PMID- 2571736 TI - Khat, toxic reactions to this substance, its similarities to amphetamine, and the implications of treatment for such patients. PMID- 2571737 TI - The characterization of beta-adrenergic receptor subtypes of the upper and lower renal pelvis in rabbits. AB - We characterized the beta-adrenergic receptors in the upper (pacemaker) and lower (nonpacemaker) regions of the rabbit renal pelvis, using radioligand binding techniques. [3H]Dihydroalprenolol was the ligand used for determining the total density of beta-adrenergic receptors. The beta-1 and beta-2 subtypes of beta adrenergic receptors were defined by inhibition of [3H]DHA binding by ICI 89,406, a beta-1 selective antagonist, and ICI 118,551, a beta-2 selective antagonist. Saturation studies with [3H]DHA showed that there was no significant difference in the equilibrium dissociation constant, KD, and the maximum number of binding sites Bmax, between the upper and lower renal pelvis. Although the inhibition constants of ICI 118,551 were smaller than those of ICI 89,406 in both the upper and lower pelvis, indicating a predominance of beta-2 receptors in both regions, the Ki values of ICI 118,551 were significantly greater in the upper than in lower pelvis. These data suggest that there are significant amounts of beta adrenergic receptors in the rabbit renal pelvis without regional differences in the total density of beta receptors and that there is a greater proportion of beta-2 subtypes in the lower than in the upper renal pelvis. PMID- 2571738 TI - Prevalence of carcinoma in situ and other histopathological abnormalities in testes of men with a history of cryptorchidism. AB - The incidence of invasive testicular cancer is increased in men with a history of cryptorchidism. Previous studies based on relatively small series indicated that the risk of carcinoma in situ of the testis also is increased in these men. In our study 500 consecutive men 20 to 30 years old, who were previously admitted to a department of surgery with the diagnosis of testicular maldescent, were asked to participate in a screening for carcinoma in situ of the testis. Of the men 300 consented to testicular biopsy. The biopsies were evaluated by light microscopy for carcinoma in situ and other histopathological abnormalities. Carcinoma in situ was diagnosed in 5 patients (1.7%, 95% confidence limits 0.5 to 3.9%). However, the true risk of carcinoma in situ might be higher, since 2 men who had been treated for testicular cancer before they were offered biopsy were excluded from the study. Advanced spermatogenesis, including the spermatid stage in all tubules, was found in biopsy specimens from only 37% of the men. In 80% of these specimens even the number of late spermatids was decreased. Thus, our study, based on a large number of testicular biopsies from an unselected group of men with testicular maldescent, provided further evidence that these men have an increased risk for carcinoma in situ of the testis. Our data combined with the results of other Scandinavian studies indicate that the true prevalence of carcinoma in situ in men with a history of cryptorchidism is approximately 2 to 3%. Additionally, we confirmed that spermatogenic function is severely impaired in maldescended gonads. Invasive testicular cancer can be prevented if the neoplasm is detected at the stage of carcinoma in situ. In our opinion the magnitude of prevalence of carcinoma in situ found in men with a history of cryptorchidism justifies that these men should be offered testicular biopsy when they reach adulthood. PMID- 2571739 TI - [Distribution and prevalence of Campylobacter pylori in the stomach]. AB - We investigated the distribution and prevalence of Campylobacter pylori in the stomach and duodenum. In this study, 500 biopsy specimens were obtained from 245 patients. In each case, biopsy specimens were taken from more than 2 sites. C. pylori was detected by culture, urease test and acridine-orange stain. C. pylori was not detected on the intestinal metaplasia, gastric cancer tissue and duodenal mucosa without gastric metaplasia. In 21% of cases, C. pylori was detected in only one site. Because of the patchy distribution of C. pylori, more than 2 biopsy specimens from different sites were needed to avoid sampling error. Detection rate of C. pylori was almost equal in antrum, angle and body as well as in male and female. H2 receptor antagonists did not affect the detection rate of C. pylori. According to the endoscopic diagnosis of the biopsied site, C. pylori was detected in 87% of gastric ulcer, 60% of duodenal ulcer (duodenal mucosa with gastric metaplasia), 73% of chronic gastritis and 62% of endoscopically normal gastric mucosa. PMID- 2571740 TI - [Clinical evaluation of serum sialyl SSEA-1 (SLX) in diagnosis of cancers]. AB - In order to evaluate the usefulness of sialyl SSEA-1 (SLX) as a tumor marker of digestive cancers, serum levels of the antigen were determined in 334 patients with malignancies and 196 patients with benign diseases. The results indicated that positivity of the antigen in sera from malignant patients was highest in pancreatic cancer (58%) and biliary tract cancer (56%). False positive incidence of SLX in sera from benign diseases was as low as 6%, revealing low false positivity. Comparison with other tumor markers such as CA19-9, CA-50, CEA and ST 439 showed that positivity of SLX was as high as that of CEA, whereas it was lower than that of CA19-9 or CA-50. On the other hand, false positivity of SLX as well as ST-439 was lowest, and accuracy of SLX was no less high than that of CA19 9 or CA-50. In sera of pancreatic and biliary tract cancer, positive incidences of CA19-9, CEA and ST-439 were 80%, 64% and 53%, respectively, and the diagnostic efficiency increased by combined assay of SLX with CA19-9 (88%), CEA (81%) and ST 439 (71%). SLX appears to be no less useful than the other recently developed carbohydrate antigens or CEA as serum tumor marker for pancreatico-biliary cancer. PMID- 2571741 TI - Effects of famotidine, a new histamine H2-receptor antagonist, on renal function. AB - The effects of famotidine on renal function were investigated. Eight healthy men (N) and 8 renal patients with varying degrees of renal failure (R.F.) participated in the trial. They were given either famotidine (40 mg) or cimetidine (800 mg) for 7 days. Cimetidine produced a significant decrease in creatinine clearance (from 131.6 +/- 12.9 to 107.7 +/- 3.4 ml/min (N), and 22.2 +/- 3.9 to 18.1 +/- 3.5 ml/min (R.F.], and increase in serum creatinine (from 0.96 +/- 0.05 to 1.09 +/- 0.04 mg/dl (N), and 3.46 +/- 0.62 to 3.86 +/- 0.51 mg/dl (R.F.], and a decrease in creatinine excretion (from 24.0 +/- 1.1 to 22.6 +/- 0.8 mg/kg/24 hr (N], respectively, although the reduction in creatinine excretion in the renal failure group was small. On the other hand, the famotidine treatment produced no significant changes in renal function in both subjects (creatinine clearance, 136.5 +/- 5.7 to 133.6 +/- 6.0 ml/min (N), and 21.9 +/- 3.6 to 20.9 +/- 4.1 ml/min (R.F.); serum creatinine, 0.98 +/- 0.02 to 0.99 +/- 0.02 mg/dl (N), and 3.24 +/- 0.43 to 3.46 +/- 0.55 mg/dl (R.F.); and urinary creatine, 25.1 +/- 1.0 to 25.3 +/- 1.0 mg/kg/24 hr (N), and 17.2 +/- 1.4 to 16.6 +/- 1.5 mg/kg/24 hr (R.F.]. There were no changes in the percent sodium excretion, or the serum and urinary beta 2-microglobulin in both groups. PMID- 2571742 TI - [Manifestation of AIDS in HIV infected homosexual males with lymphadenopathy syndrome]. AB - 26 homosexual men with HIV-infection and lymphadenopathy syndrome were examined between 1982 and 1984 and prospectively observed over a period of 58 months (median, range 53-69 months). 16 (61.5%) developed full blown AIDS within a median period of 41 months (23-63 months). 12 of them died 50 months (median, range 29-67 months) after the first examination. 10 patients showed no symptoms of AIDS after a median observation time of 57 months (54-63 months). A panel of clinical symptoms, signs of concomitant infections, chemical and immunological analyses, including a lymphnode biopsy were correlated to the course of the disease. A decreased proportion of CD4-cells and a lower CD4/CD8 ratio in the lymphnode suspensions at the first examination in patients with disease progression were the only parameter which correlated significantly with the development of AIDS. PMID- 2571743 TI - Early diagnosis of multiple endocrine neoplasia type IIa. AB - We report on incidental findings during family screening of two kindreds with multiple endocrine neoplasia type IIa. Pheochromocytoma and medullary thyroid carcinoma of considerable size were detected. The results underline the importance of early diagnosis of the syndrome, since the afflicted may be almost or wholly asymptomatic. High resolution chromosome banding studies were carried out in both families, but no abnormality was found. Linkage analysis using DNA markers closely related to the chromosomal locus at chromosome 10 was carried out and was positive in two asymptomatic offspring of one family, whereas the markers were not informative in a second family. We recommend early linkage analysis for establishing the genetic status in offspring of multiple endocrine neoplasia type IIa families to identify for further screening those who are predicted to be gene carrier. PMID- 2571744 TI - Distribution of injected monoclonal antibody in lymphoma-infiltrated spleen. AB - Specific and non-specific antibody was injected into mice with lymphoma infiltrated spleens in order to assess the distribution with both time and with extent of tumour infiltration. The specific antibody (MRC OX7) bound to the Thy 1.1 antigen on the lymphoma cells (A120). There was evidence that the diffusion of MRC OX7 deeply into the tumour was hindered by competitive binding to the tumour cells immediately surrounding the supplying blood vessels. In some circumstances this situation persisted to 24 h, after which time the antibodies were being cleared from the host in significant amounts. PMID- 2571745 TI - [Problems of the two-stage restoration of flexor tendon injuries. I. Current results. Biological and functional factors affecting healing]. AB - After describing the morphological and biological relations in the intact human flexor tendon and tendon sheat unit authors deal in details with the severe injuries of the flexor tendons inside the tendon sheat, having a bad prognosis. The results of the previous experimental investigations, and among them their own results, are reviewed. The success of the two-phase tendon transplantations in this kind of injury is determined, according to their opinion, beside the known clinically related factors, by biological and functional factors. Further examination of the complicated reparative processes with the use of modern methods of investigation is planned. The present work is ment for introduction of these experiments. PMID- 2571746 TI - [Problems of the two-stage restoration of flexor tendon injuries. II. Light- and scanning electron microscopy studies of flexor tendon sheaths from hens and from humans]. AB - Authors examined the normal flexor tendon/tendon sheat unit of the toes of the hen and in the zone Verdan 2 of the human hand with light and scanning electron microscope. The fine structure of the synovial membrane was studied with special care. It was found, in accordance with some previous literary data, that the morphology of the hen's and human tenosynovium resemble to each other. The structures of the parietal visceral synovial membranes are described and compared, and the mechanism of the secretion of the synovial fluid is discussed. The results of the present investigations may serve, over the description of the normal anatomical relations, as a comparison to other experiments of two-phase tendon transplantations with the formation of pseudo tendon sheats and also for clinical practice. PMID- 2571747 TI - [Possibilities of restoration of fresh ligament injuries of the knee]. AB - Author briefly summarizes the changes in the aspects and diagnostics of the ligament injuries of the knee. He deals with the methods of treatment used at his Department and with the operative procedures and describes in details the types of injuries of the single parts and the possibilities of reconstruction. His unambigous opinion is that in fresh ligament injuries the best result can be expected from the early complete reconstruction. In spite of the fact that "restitutio and integrum" can be reached only in the minority of the cases, the results are better than in conservative treatment or in ligamentous plasties. PMID- 2571748 TI - [Results of treatment of infected osteosynthesis cases]. AB - At the author's Department 70 infected osteosyntheses were treated. The preliminaries of infection were in 48 cases open, and in 22 cases closed fractures. The original stabilisation was performed in 48 injuried with plates, in 12 cases with intramedullary nails and in 10 other patients with various methods. In 12 cases healing of the infection was reached in a short time. In 58 cases the infection became chronic. In 34 cases the fractures healed in spite of persistant infection, in 24 cases infected pseudarthrosis developed. To reach healing or the definite state frequently more operations were needed. Finally it succeeded to heal both the infection and the fracture in 62 patients. Fistula, resistant to treatment remained in 7 patients. PMID- 2571749 TI - [Restoration of movement after irreparable nerve damage caused by injury]. AB - Authors made a late follow-up of their motion improving operations, performed on the Department of Traumatology of the I Department of Surgery, Pesci University of Medical Science. The indication and technique of these operations in case of paralysis of the opponens, interosseal-lumbrical muscles and of the extensors of the hand are described in details. PMID- 2571750 TI - [Management of calcaneus fractures]. AB - Author describes the diagnostics and the classification of the types of the dislocated calcaneal fractures. A closed reduction of these fractures with a special apparatus and fixation with percutaneous screws in performed. No postoperative external fixation is used, early active motion therapy is introduced and early weight bearing is permitted. The results, that are better than in the previous methods of treatment, are presented in 225, similarly treated and controlled cases. PMID- 2571752 TI - [Chiari's pelvic osteotomy in the management of Perthes disease]. AB - Authors deal, on the basis of an analysis of 3 own cases and of the literary data, with the indication of Chiari's osteotomy in Perthes' disease. It is stated that the operation is to be performed over the age of 8-9 years, in severe lesion of the femoral head, and if the femoral head is so much flattened, widened and extruded from acetabulum that it can not be centralized with varisation. The operation promotes the rebuilding of the femoral head epiphysis, the biomechanical relations are improved and the early arthrosis can be prevented. PMID- 2571753 TI - [Possibilities of using preserved tendon in hand surgery. Review of the literature]. AB - Authors review the possibilities of the use of conserved tendon, especially on the field of flexor tendon reconstruction--a problem that can not be considered to be solved entirely. The requirements of the conserved tendon are reviewed, the question of the antigenicity is emphasized and the process of tendon regeneration, using conserved tendon, is described. The various procedures of conservation e.g. freeze drying and different chemical methods are compared and the criteria of the use of conserved tendon are discussed. The possibilities of the use of conserved tendon are mentioned, several questions, requiring further investigations, are posed. A wider use of conserved tendon in the future is recommended. PMID- 2571754 TI - [Limb-preserving resection operations in the management of bone tumors of the shoulder region]. AB - Authors describe the frequency of bone tumours in the shoulder region and the anatomical characteristics of this region, that are important, regarding tumour surgery. Six cases are reported in whom resection was performed for malignant, semimalignant and benign bone tumours. In their material one primary and two secondary chondrosarcomas, one Ewing's sarcoma, one osteoclastoma and one benign chondroblastoma were found. Follow-up range was 3-7 years. In two patients partial and total scapulectomy was performed, in the later the proximal end of the humerus was resected only, in three of them endoprosthesis was given, and in one case the missing bone was replaced with a fibular graft. The function of the limbs was in every case, even in those in which the replacement of the bone segment was not carried out, adequate. Metastases developed in three patients, two of them were lost, one is alive. The rest of the patients was free of tumour at the follow-up. PMID- 2571751 TI - [Results of replantation of limbs]. AB - Authors describe 7 cases of limb replantation in their material. The field of indication is reviewed, the early postoperative complications and the possibilities of their treatment are discussed. The means of reaching the possible best results are examined. The ways of performing the possible optimal limb replantation are scheduled on the basis of their own experiences. PMID- 2571755 TI - [Multiple drug resistance. Biological and clinica implications]. PMID- 2571756 TI - [The congress on public health. Fixed care compensation in hospitals in the USA are not a threat against new technologies]. PMID- 2571757 TI - [A consensus conference in Brugge: guidelines for prevention of psychosis in schizophrenia]. PMID- 2571758 TI - Long-term glycaemic control and diabetic retinopathy. AB - 216 patients with insulin-dependent diabetes mellitus were studied by retinal photography, and the absence or presence of retinopathy was related to the mean of serial glycosylated haemoglobin measurements (mean HbA1) carried out every 3 months during the previous 6 years. 122 patients had no diabetic retinopathy, 86 had background retinopathy, and 8 proliferative retinopathy. Mean HbA1 levels showed a strong correlation with increasingly severe grades of retinopathy, even when differences in duration of diabetes were taken into account. Proliferative retinopathy was seen only in patients with mean HbA1 above 10%. These results support the view that the development of diabetic retinopathy is related to long term glycaemic control and emphasise the desirability, and possible benefit, of achieving control as close to normal as is possible for each individual patient. PMID- 2571759 TI - Lack of efficacy of pyrimethamine prophylaxis in pregnant Nigerian women. AB - To evaluate the efficacy of pyrimethamine on the blood stage (suppressive prophylaxis) and liver stage (causal prophylaxis) of Plasmodium falciparum in pregnant women, in vivo and in vitro field studies were conducted in Ilorin, Nigeria, from Jan 1 to June 30, 1988. For pregnant women with P falciparum infections who received 25 mg of pyrimethamine weekly for suppressive prophylaxis, 67% (59/88) of in vivo and 60% (6/10) of in vitro tests showed pyrimethamine resistance. A second group of parasitaemic and parasite-free pregnant women was enrolled to evaluate the efficacy of pyrimethamine as a primary tissue schizonticide; after receiving a curative dose of chloroquine (25 mg/kg), half the women were given 25 mg of pyrimethamine weekly and half received no prophylaxis. Parasitologic failure rates did not differ between the pyrimethamine-treated (8/34) and the control (11/37) groups during the 16-week follow-up. Thus, pyrimethamine is not effective for suppressive or causal prophylaxis in pregnant women in Ilorin. PMID- 2571760 TI - Aortic aneurysms as late sequelae of above-knee amputation. AB - The infrarenal aorta was examined by ultrasound in 329 men who had lost a leg in World War II and in 702 war veterans. The groups were similar in terms of age and risk factors for arteriosclerosis. Abdominal aortic aneurysms were found in 5.8% of the amputees compared with 1.1% of the non-amputees. Unilateral flow reduction after leg amputation causes an asymmetrical flow pattern at the aortic bifurcation, and this is probably the main cause of late damage to the aorta. PMID- 2571761 TI - The role of bronchial musculature. PMID- 2571762 TI - Thrifty genotype rendered detrimental by progress? PMID- 2571763 TI - Salted fish and nasopharyngeal carcinoma. PMID- 2571765 TI - Not all upside down. PMID- 2571764 TI - Ultrasound and bone fragility. PMID- 2571766 TI - Spontaneous pneumothorax. PMID- 2571767 TI - Stones and kilograms. PMID- 2571768 TI - Prediction of serious post-infarction cardiac arrhythmias. AB - Simple clinical findings suggestive of a large myocardial infarction identify most patients at risk from serious post-infarction arrhythmias (defined as sustained ventricular tachycardia and sudden death not associated with reinfarction). However, similar findings are seen in many other patients who remain free of complications. The presence of late potentials and frequent ventricular ectopics on long-term electrocardiographic monitoring add to prognostic accuracy in patients with clinical complications, with a predictive accuracy of nearly 90% when all three features are present. PMID- 2571769 TI - Half-life of HBs antibody after hepatitis B vaccination: an aid to timing of booster vaccination. AB - Antibody to hepatitis B virus surface antigen was measured in 48 paired sera from healthy medical staff who had been vaccinated against hepatitis B. The rate of antibody decline seemed unrelated to the height of the post-vaccination antibody titre and varied widely: even after correction for error of measurement, half lives ranged from 40 to 280 days (mean 150, SD 52). For maintenance of protection, the recommended date for revaccination should be based on both the post-vaccination antibody level and the half-life, determined individually on completion of vaccination. PMID- 2571770 TI - Outbreak of botulism in north west England and Wales, June, 1989. AB - The clinical features of 27 patients identified in an outbreak of botulism in Lancashire, England, and North Wales are reviewed. All but 1 of the patients (age range 14 months to 74 years) were admitted to hospital: 12 were treated in intensive care units, and 8 received positive pressure ventilation. 1 patient died with an aspiration pneumonia. The clinical presentations contained several unusual features, with evidence of segmental demyelination in some patients and drowsiness, sore throats, and fever in others. The widely dispersed source of intoxication with patients presenting singly to several hospitals added to the difficulties of diagnosis. Successful clinical management depends on full and early recognition both of the dangers of impaired oropharyngeal function and of the rapid neurological changes in botulism. PMID- 2571771 TI - Ultrasonography and scintigraphy in liver disease in developing countries. A retrospective survey. AB - In a study of the value of hepatic ultrasonography (US) and scintigraphy (SG) in detecting liver disease in developing countries 425 US scans and 304 SG scans of patients with focal or diffuse liver disease or normal livers were reviewed. The accuracy of both US and SG in distinguishing between normal and diseased livers was low (68% and 74%, respectively). Both techniques did better at detecting focal than diffuse liver disease; the sensitivity of US and SG in focal and diffuse disease was 88% and 92%, and 27% and 54%, respectively. The specificity of both procedures was high for both types of liver disease (91-96%). Overlap between US features of amoebic liver abscess, hepatocellular carcinoma, and metastatic carcinoma resulted in a correct final diagnosis being made in only 81% of patients with amoebic liver abscess, 29% with hepatocellular carcinoma, and 43% of patients with metastatic carcinoma who had a US scan. This study indicates that these techniques are neither accurate in detecting diffuse liver disease nor capable of determining the cause of diffuse liver disease. When diffuse parenchymal liver disease is suspected biopsy would be needed. Although the accuracy of both imaging modalities in detecting focal disease is high, overlap between the US features of the common causes of space-occupying lesions may result in an incorrect final diagnosis in some cases. In consequence, biopsy or aspiration might be required. PMID- 2571772 TI - Rationing and other perils for providers and consumers. PMID- 2571773 TI - Risk factor for myocardial infarction in transurethral resection of prostate? PMID- 2571774 TI - Octreotide. PMID- 2571775 TI - Post-exposure prophylaxis for hepatitis B. PMID- 2571776 TI - Low-dose hepatitis B vaccine. PMID- 2571777 TI - Susceptibility of human herpesvirus 6 to acyclovir. PMID- 2571778 TI - Temperature, labour, and epidural analgesia. PMID- 2571779 TI - In praise of the traditional birth attendant. PMID- 2571780 TI - Increased male births associated with placenta praevia. PMID- 2571781 TI - Maternal mortality. PMID- 2571782 TI - Sex ratio and in-vitro fertilisation. PMID- 2571783 TI - Aerosolised pentamidine and public health. PMID- 2571784 TI - Pentamidine and hypoglycaemia. PMID- 2571785 TI - Skin surface microscopy. PMID- 2571786 TI - Acute psychosis following mefloquine prophylaxis. PMID- 2571788 TI - Angle on disability. PMID- 2571787 TI - Short-term metabolic effects of pamidronate in patients with prostatic carcinoma and bone metastases. PMID- 2571789 TI - Late abortions. PMID- 2571790 TI - Torture in Turkey. PMID- 2571791 TI - World Medical Association and South Africa. PMID- 2571792 TI - Clostridium difficile infection in health-care workers. PMID- 2571793 TI - Action of nitroprusside in multidrug-resistant hypertension. PMID- 2571794 TI - Carbamazepine update. PMID- 2571795 TI - Interaction between fluconazole and cyclosporin. PMID- 2571796 TI - Food-based oral rehydration salt solution for acute childhood diarrhoea. PMID- 2571797 TI - HLA subtypes and meningococcal disease. PMID- 2571798 TI - Detecting fetal cells in maternal circulation. PMID- 2571800 TI - Air bubbles in insulin pens. PMID- 2571799 TI - Disassociation between debrisoquine hydroxylation phenotype and genotype among Chinese. PMID- 2571801 TI - Detection of malaria antigen in cerebrospinal fluid by counter-current immunoelectrophoresis. PMID- 2571802 TI - Prenatal diagnosis of triose phosphate isomerase deficiency. PMID- 2571803 TI - Severe facial oedema and glossitis associated with mianserin. PMID- 2571804 TI - Let sleeping giardia lie. PMID- 2571805 TI - Lignocaine prophylaxis for reperfusion arrhythmias during treatment with streptokinase in acute myocardial infarction. PMID- 2571806 TI - Possible role of autonomic afferents in treatment of postoperative hiccups. PMID- 2571807 TI - Short-bowel radiation injury. PMID- 2571808 TI - Addison's disease and primary antiphospholipid syndrome. PMID- 2571809 TI - Parkinsonism due to n-hexane exposure. PMID- 2571810 TI - Familial occurrence of opioid-induced biliary colic. PMID- 2571811 TI - Sodium balance in infants with severe congestive heart failure. PMID- 2571812 TI - Intensive treatment of multiple myeloma and criteria for complete remission. AB - 50 previously untreated patients with multiple myeloma received two-phase treatment: repeated cycles of 4 day infusion with vincristine, doxorubicin, and methylprednisolone (VAMP) followed by high-dose melphalan (HDM), with autologous bone marrow transplantation where possible. The overall response rate was 74% (37/50), with 25 patients (50%) achieving complete haematological and biochemical remission. These remissions were associated with a good quality of life as measured by performance status, pain grade, and the reversal of humoral immunosuppression. 6 patients died during the VAMP phase and there was 1 death related to HDM. The achievement of complete remission, as defined here, in such a high proportion of patients is exceptional and may represent a useful advance in the management of myeloma. PMID- 2571813 TI - Infusion of vincristine and doxorubicin with oral dexamethasone as first-line therapy for multiple myeloma. AB - 32 previously untreated patients with multiple myeloma received vincristine, doxorubicin ('Adriamycin'), and dexamethasone (VAD) as first-line therapy. The overall response rate was 84%, with 28% of all patients entering complete remission. Response was rapid, with near-maximum response occurring after two courses of treatment and rapid improvement in bone-marrow function. Median response duration was 18 months and this seemed to be unaffected by initial prognostic criteria or by degree of remission achieved. The projected median survival was 44 months, with 75% of all patients and 83% of responders being alive at 2 years. Side-effects due to steroids were common, but there was only 1 treatment-related death. The high response rate and lack of toxicity offer an advantage over other forms of initial treatment, although other strategies will be necessary to prolong the duration of response. PMID- 2571814 TI - Demonstration of mother-to-infant transmission of hepatitis B virus by means of polymerase chain reaction. AB - To investigate the failure of vaccines to prevent mother-to-infant transmission of hepatitis B virus (HBV), serum, cord blood, and colostrum samples from eleven mothers, known to be carriers of hepatitis B surface antigen, and their infants were examined by means of a highly sensitive polymerase chain reaction (PCR) method. HBV-specific DNA was detected in ten maternal serum samples, eight samples of colostral whey, eight samples of colostral cells, and one cord blood sample. Four infants of mothers with HBV-DNA-positive colostrum showed low responsiveness to hepatitis B vaccine. The infant whose cord blood was positive for HBV DNA showed low responsiveness to hepatitis B vaccine and subsequently became an HBV carrier. These results suggest the need for further study to evaluate whether breastfeeding is advisable for HBV carriers. PMID- 2571815 TI - Effect of psychosocial treatment on survival of patients with metastatic breast cancer. AB - The effect of psychosocial intervention on time of survival of 86 patients with metastatic breast cancer was studied prospectively. The 1 year intervention consisted of weekly supportive group therapy with self-hypnosis for pain. Both the treatment (n = 50) and control groups (n = 36) had routine oncological care. At 10 year follow-up, only 3 of the patients were alive, and death records were obtained for the other 83. Survival from time of randomisation and onset of intervention was a mean 36.6 (SD 37.6) months in the intervention group compared with 18.9 (10.8) months in the control group, a significant difference. Survival plots indicated that divergence in survival began at 20 months after entry, or 8 months after intervention ended. PMID- 2571816 TI - DNA technology and rapid diagnosis of infection. PMID- 2571817 TI - Non-Q-wave myocardial infarction. PMID- 2571819 TI - Child health in the inner cities. PMID- 2571818 TI - What price MRC initiatives now? PMID- 2571820 TI - Psychosocial intervention and the natural history of cancer. PMID- 2571821 TI - Hypophosphataemic rickets: final height and clinical symptoms in adults. AB - The heights and symptoms of 52 patients, aged at least 18 years, with X-linked hypophosphataemic rickets were analysed retrospectively; 47 had been seen as children and 5 were adult at their first examination. 2 patients were lost to follow-up. 3 patients had died but their adult heights were known. There was no evidence that any form of treatment (ie, vitamin D in high doses, vitamin D plus phosphate supplements, or calcitriol plus phosphate) had any effect on adult height, symptoms, or alkaline phosphatase levels. There was a negative relation between adult height and the number of osteotomies undergone. The complications of treatment, such as renal failure, which occurred secondary to vitamin D intoxication in 3 patients in their twenties, may outweight any possible benefits. Until a treatment is established as effective in controlled trials, it may be better that these patients remain untreated. PMID- 2571823 TI - Visuospatial neglect: underlying factors and test sensitivity. AB - Visuospatial neglect, a frequent consequence of unilateral (usually right hemisphere) stroke, is associated with poor functional recovery and in many patients is resistant to remedial treatment. Studies of the nature and prevalence of the disorder have been hindered by problems of definition and assessment. In this study 80 unselected stroke patients were assessed for the presence and severity of neglect on the behavioural inattention test. The six subtests of this battery all intercorrelated highly, and a subsequent factor analysis showed that all tests loaded significantly on one underlying factor. The construct of neglect as defined by performance on the battery is therefore robust. Nonetheless, the individual tests differed substantially in their sensitivity. Star cancellation was the most sensitive measure of neglect and correctly diagnosed all patients whose aggregate score on the full battery fell below that of the control population. PMID- 2571822 TI - How accurate is computer-aided diagnosis? AB - A computer program for diagnosis of acute abdominal pain (CAD-A) was compared with an amended program (DIAG) and with clinical performance. The programs were tested at three Scottish hospitals, representing a range of practice, in 6962 cases. Accuracy of CAD-A lay in the range 48-59%, and of DIAG 56-62%. Clinical accuracy was constant at 65%. These figures suggest that computer systems based on Bayes' formula have no useful role in the diagnosis of acute abdominal pain. PMID- 2571824 TI - The relief of communication. PMID- 2571825 TI - Progression of osteoarthritis and drug treatment. PMID- 2571826 TI - Plasma exchange and immunosuppression in the stiff man syndrome. PMID- 2571827 TI - Non-mosaic trisomy 16 confined to villi. PMID- 2571828 TI - Hymenoptera stings and beta-blockers. PMID- 2571829 TI - Intrathecal baclofen for cerebral spasticity. PMID- 2571830 TI - Benzodiazepine prescribing. PMID- 2571831 TI - Ergot and idiopathic sclerosing encapsulating peritonitis. PMID- 2571832 TI - Trends in brain cancer. PMID- 2571833 TI - Achilles tendon rupture. PMID- 2571834 TI - Infant mortality in offspring of Australian doctors. PMID- 2571835 TI - Effect of various anti-ulcer drugs on rapid urease test for Campylobacter pylori infection. PMID- 2571836 TI - Smallpox vaccination for investigators. PMID- 2571837 TI - Postoperative pyrexia, atropine, and malaria. PMID- 2571838 TI - Extrapyramidal signs in dementia of Alzheimer type. PMID- 2571839 TI - Parkinsonism and Down syndrome. PMID- 2571840 TI - Motoneuron disease and solvent and glue exposure. PMID- 2571841 TI - Predictive testing for Huntington's disease with linked DNA markers. PMID- 2571843 TI - Mature entrants to medicine. PMID- 2571842 TI - Informing patients. PMID- 2571844 TI - Local application of hyperosmolar glucose solution in tubal pregnancy. PMID- 2571845 TI - Raised plasma aluminium in an infant on antacid. PMID- 2571846 TI - Smoking and cerebral ischaemia. PMID- 2571847 TI - Steroid-induced psychosis. PMID- 2571848 TI - Spontaneous lymphocyte proliferation in symptom-free HTLV-I positive Jamaicans. PMID- 2571849 TI - Prevalence of genetic haemochromatosis in diabetic patients. PMID- 2571850 TI - Intrauterine surgery under intravenous sedation as an outpatient alternative to hysterectomy. PMID- 2571852 TI - Identification of a new antinuclear antibody by immunoblotting. PMID- 2571851 TI - Hazards of interferon therapy for HBV-seronegative chronic hepatitis. PMID- 2571853 TI - Mumps vaccine meningitis. PMID- 2571854 TI - Lactulose as an anti-endotoxin in liver transplantation. PMID- 2571855 TI - Acute hypertension mimicking phaeochromocytoma as main presenting feature of acute intermittent porphyria. PMID- 2571856 TI - Risk factors for melanoma. PMID- 2571857 TI - Screening and treatment to prevent post abortion chlamydial pelvic infection. PMID- 2571858 TI - Photocopier's papillitis. PMID- 2571859 TI - Frauds on the public. PMID- 2571860 TI - Effect of graft perfusion with two CD45 monoclonal antibodies on incidence of kidney allograft rejection. AB - In a blind trial, 77 patients were randomised to receive first cadaver kidney allografts that had been perfused either with a pair of CD45 monoclonal antibodies (mAbs), in an attempt to reduce the immunogenicity of passenger leucocytes, or with control human albumin solution. No complications of mAb perfusion were observed. Patient and allograft survival were similar in both groups. Rejection episodes were recorded in 7 (18%) of the patient with mAb perfused allografts compared with 24 (63%) of the controls. PMID- 2571861 TI - Association of painful peripheral neuropathy in AIDS with cytomegalovirus infection. AB - 12 AIDS patients with a clinically, electrophysiologically, and pathologically homogeneous painful peripheral neuropathy had a significantly higher rate of any or active clinical cytomegalovirus (CMV) infection than did a group of 30 AIDS controls. In 9 patients this syndrome began in close temporal association with clinical infection with CMV. Painful peripheral neuropathy in AIDS may be caused by a CMV dorsal root ganglionitis. PMID- 2571862 TI - Influence of plaque configuration and stress distribution on fissuring of coronary atherosclerotic plaques. AB - To find out the types of atherosclerotic plaques that fissure and where they fissure, plaques from 85 patients who had died from coronary thrombosis were examined histologically. 67 of the plaques contained an eccentric pool of extracellular lipid in the intima; 42 (63%) of these plaques had fissured at the junction of the plaque cap with the more normal intima, and the other 25 (37%) had torn through the centre of the cap. Computer modelling of different forms of plaque showed that at systole eccentric pools of lipid concentrated stress on the plaque cap, especially near the edge of the plaque. When the lipid pool occupied less than 15% of the vessel circumference, and when the plaque cap was less stiff than the adjacent normal intima, the point of maximum stress was over the centre of the plaque. Computer modelling also showed that the distribution of circumferential tensile stress across the intima was radically altered by atherosclerotic plaques. Regions of high circumferential stress correlated well with the site of intimal tears found at necropsy. The histological findings showed that site of tearing was influenced by variation in the mechanical strength of cap tissue due to focal accumulation of foam cells. Focal weak points in the cap would explain tears which were not at the point of maximum stress. PMID- 2571863 TI - Fish intake and arterial wall characteristics in healthy people and diabetic patients. AB - The relation between fish consumption and indices of arterial wall pathology was investigated in 31 healthy subjects and in 22 patients with non-insulin-dependent diabetes mellitus (NIDDM). Arterial compliance in non-fish eaters, as measured by doppler ultrasonography, was significantly lower than that in fish eaters in the healthy group, in the NIDDM group, and in the two groups combined. In non-fish eaters an increase in proximal resistance at the common femoral artery and in the posterior tibial artery was significant only in the combined groups and in healthy subjects, respectively. The results support the hypothesis that fish consumption may be important for better arterial wall characteristics. PMID- 2571864 TI - Responses to human and porcine insulin in healthy subjects. AB - In a double-blind randomised study, eight healthy men received equimolar amounts of human or porcine insulin by infusion (50 mU/kg per hour). Insulin potencies, in terms of the amount of glucose infused to maintain euglycaemia, were almost identical. Hypoglycaemia (blood glucose concentration 2 mmol/l or below) was then induced and the symptoms and hormonal counter-regulatory responses were recorded. The number of sympathoadrenergic (but not neuroglycopenic) symptoms was significantly greater with porcine than with human insulin, as was the integrated noradrenaline response. Glucagon, growth hormone, cortisol, and adrenaline responses were similar for the two insulins. PMID- 2571865 TI - Menstrual influence on surgical cure of breast cancer. AB - In a retrospective study of 44 premenopausal women who underwent resection of a primary breast cancer and were followed for 5 to 12 years, disease recurrence and metastasis were more frequent and more rapid in women who had been operated upon during the perimenstrual period (days 0-6 and 21-36 of the menstrual cycle). By multivariate analysis, the time of resection in relation to the menstrual cycle is an independent predictor of the likelihood of future metastatic disease. Patients who underwent resection during the perimenstrual period had a more than quadrupled risk of recurrence and death compared with women operated upon during days 7 to 20 of the menstrual cycle. PMID- 2571866 TI - Simple nebuliser modification to enhance alveolar deposition of pentamidine. PMID- 2571867 TI - Tumour cell vaccines: has their time arrived. PMID- 2571868 TI - Abnormal blood clotting after head injury. PMID- 2571869 TI - Peptides into the coronary arteries. PMID- 2571870 TI - Prognostic factors in childhood rhabdomyosarcoma. PMID- 2571871 TI - X rays for the developing world. PMID- 2571872 TI - Viral gastroenteritis aboard a cruise ship. AB - A 32-nm small round structured virus (SRSV), possibly related to the Snow Mountain agent (SMA), was implicated as the cause of recurrent outbreaks of gastroenteritis on a cruise ship. There was no identifiable relation to food or water consumption, but the risk of gastroenteritis among passengers who had shared toilet facilities was twice that of those who had a private bathroom and the rate of illness was related to the number of passengers sharing a communal restroom (ie, with one or more toilets): contaminated bathrooms may be an important vehicle for person-to-person spread of this enteric agent. In each cabin, index patients who had vomited in their cabins were more likely to have had cabinmates who subsequently became ill than were index patients who had not vomited. These epidemiological findings implicate vomitus in the transmission of viral gastroenteritis and they are consistent with the transmission of viral agents by airborne droplets or person-to-person contact. New strategies for prevention of viral gastroenteritis should include protection against environmental contamination by viruses in airborne droplets or vomitus. PMID- 2571873 TI - Efficacy of tests used to monitor rheumatoid arthritis. AB - The relative efficacy of clinical and laboratory tests used to monitor disease activity in rheumatoid arthritis was determined by consensus analysis in a study of 21 patients treated for 6 months. Erythrocyte sedimentation rate (ESR), which is influenced by the anaemia of chronic disease and by variation in the blood concentration of acute-phase proteins, was the most effective single test. ESR was a better guide to disease severity than measurement of plasma viscosity, serum C-reactive protein, and serum orosomucoid--tests that reflect the blood concentration of acute-phase proteins only. Clinical tests performed poorly unless combined with laboratory data in a numerical index. Consensus analysis can be used to assess the efficacy of clinical and laboratory tests and to identify redundant tests. PMID- 2571875 TI - Prenatal detection of major cystic fibrosis mutation. PMID- 2571874 TI - Tubal patency after local methotrexate injection for tubal pregnancy. AB - Tubal patency was investigated by hysterosalpingography in 21 of 37 patients with unruptured tubal pregnancy treated by local methotrexate injection at laparoscopy. 18 of the 21 patients had bilateral tubal patency, and the only tube of a patient with a single fallopian tube was also patent. 6 subsequent intrauterine pregnancies have so far been recorded. Local methotrexate injection into the tubal pregnancy may provide an efficient and safe alternative to surgery in early unruptured ectopic pregnancy. PMID- 2571876 TI - First-trimester prenatal diagnosis of cystic fibrosis by direct gene probing. PMID- 2571877 TI - Carrier screening for cystic fibrosis. PMID- 2571878 TI - Treatment of lichen planus with temarotene. PMID- 2571879 TI - Propofol and intensive care. PMID- 2571880 TI - 5-HT receptors and arteriovenous shunts. PMID- 2571881 TI - Cyclosporin in idiopathic steroid-resistant membranous glomerulonephritis. PMID- 2571882 TI - Diabetes mellitus misdiagnosed as AIDS. PMID- 2571883 TI - Multiple drug reactions in a patient with AIDS. PMID- 2571884 TI - Female-to-male transmission of HIV-1. PMID- 2571885 TI - Endoscopic sphincterotomy. PMID- 2571886 TI - No neuroblastoma in Zaire. PMID- 2571887 TI - Cohort effect on HTLV-I seroprevalence in southern Japan. PMID- 2571888 TI - Increased brain 3-hydroxykynurenine in Huntington's disease. PMID- 2571889 TI - Carbamazepine update. PMID- 2571890 TI - Limited significance of null results. PMID- 2571891 TI - Hungerstrikes in Morocco. PMID- 2571892 TI - Beware of the cow. PMID- 2571893 TI - Informing patients about clinical disagreement. PMID- 2571894 TI - Hyperlexia in Prader-Willi syndrome. PMID- 2571895 TI - Aldehyde dehydrogenase-2 deficiency and articular collagen. PMID- 2571896 TI - Weight in infancy and death from ischaemic heart disease. PMID- 2571898 TI - Kikuchi's disease with liver dysfunction presenting as fever of unknown origin. PMID- 2571897 TI - Borrelia burgdorferi infection and Guillain-Barre syndrome. PMID- 2571899 TI - Benzodiazepine withdrawal. PMID- 2571900 TI - Hepatitis C virus and sexual transmission. PMID- 2571901 TI - Intrafamilial transmission of hepatitis C virus. PMID- 2571902 TI - Anti-phospholipid antibody syndrome and pre-eclampsia. PMID- 2571903 TI - Pancreatic transplantation with venous portal drainage. PMID- 2571904 TI - Improved primary renal allograft survival on cyclosporin limited to women with previous pregnancies. PMID- 2571905 TI - Bolus alteplase. PMID- 2571906 TI - Manual lymph drainage in nephrotic syndrome during pregnancy. PMID- 2571908 TI - Death in the dentist's chair. PMID- 2571907 TI - Enlarged salivary glands and Sjogren's syndrome. PMID- 2571909 TI - The Dutch soft-drug policy. PMID- 2571910 TI - Misuse of antidiarrhoeal medicines. PMID- 2571911 TI - Protective effect of colchiceine against acute liver damage. AB - Pretreatment of rats with colchiceine (10 micrograms/day/rat) for seven days protected against CCl4-induced liver damage. CCl4 intoxication was demonstrated histologically and by increased serum activities of alanine amino transferase (ALT), alkaline phosphatase (Alk. Phosph.) gamma glutamyl transpeptidase (GGTP), bilirubins and decreased activity of glucose-6-phosphatase (G-6Pase). Furthermore, an increase in liver lipid peroxidation and a decrease in plasma membrane GGTP and Alk. Phosph. activities were found. Colchiceine increased 1.5 fold the LD50 of CCl4 and prevented the release of intracellular enzymes as well as the decrease in GGTP and Alk. Phosph. activities in plasma membranes. It also completely prevented the lipid peroxidation induced by CCl4 and limited the extent of the histological changes. PMID- 2571912 TI - Effect of typical and atypical antipsychotic drugs on 5-HT2 receptor density in rat cerebral cortex. AB - The effect of acute treatment with seven atypical antipsychotic drugs and four typical antipsychotic drugs on serotonin2 (5-HT2) receptor binding sites in rat cerebral cortex was studied. Among the atypical antipsychotic drugs examined, clozapine, fluperlapine, RMI-81582 and setoperone decreased the density of 5-HT2 receptors, but ticspirone, amperozide and melperone did not. None of the drugs affected the Kd value. Among the typical antipsychotic drugs, loxapine decreased Bmax and increased the Kd of 5-HT2 receptor binding sites, whereas chlorpromazine and cis-flupenthixol had no effect. Clothiapine, a typical antipsychotic drug of the same chemical class as clozapine, decreased Bmax without increasing Kd. The downregulation of 5-HT2 receptor binding sites following a single injection of clozapine, 20 mg/kg, remained almost unchanged during the first 72 hrs and was still significantly decreased for up to 120 hrs. There was no relationship between the affinity for the downregulation of rat cortical 5-HT2 receptor binding site and 5-HT2 receptor density. Coadministration of the D1 dopamine agonist, SKF-38393, did not affect the clozapine-induced downregulation. It is suggested that rapid and prolonged downregulation of 5-HT2 receptor sites is characteristic of some but not all atypical antipsychotic drugs and is not specific to atypical antipsychotic drugs. Dibenzo-epines (clozapine, loxapine, amoxapine, chlothiapine) consistently downregulate 5-HT2 receptors in frontal cortex after acute treatment. PMID- 2571913 TI - RFLP-based genetic map of Phanerochaete chrysosporium ME446: lignin peroxidase genes occur in clusters. AB - We describe the construction of an RFLP-based genetic map of the white rot fungus Phanerochaete chrysosporium ME446. The map is deduced from the allele distributions of 38 RFLP markers in a test set of 53 meiotically derived haploid recombinants of this strain. The map includes a cellulase gene, a matingtype locus and a family of lignin peroxidase (and related) genes that are arranged in two unlinked clusters. PMID- 2571914 TI - Genetic factors influencing lignin peroxidase activity in Phanerochaete chrysosporium ME446. AB - Haploid recombinant progeny of Phanerochaete chrysosporium ME446, genome compositions of which had been defined by RFLP-mapping, vary in their idiophasic behaviour. This allowed us to formulate a model of the sequence of idiophasic activities. One component of this variation, the amount of lignin peroxidase activity, is independent of the allele distributions of the lignin peroxidase gene clusters, but correlates with the allele distribution of another locus. This locus appears to control the spread of the lignin peroxidase-active state within the idiophasic mycelial mat and may be the mating-type locus. The successful determination of linkage relied on analysis of chromosome intervals rather than linkage to single markers; this approach should be generally useful for analysing quantitative characters by RFLP mapping. PMID- 2571915 TI - Regulation of the phase switch controlling expression of type 1 fimbriae in Escherichia coli. AB - The expression of Escherichia coli type 1 fimbriae is phase-variable i.e. the bacterial cell is either fimbriated or non-fimbriated. The transition from one state to the other is caused by the change in configuration of an invertible DNA segment harbouring the promoter of the fimA gene. The position of this phase switch is controlled by two proteins, FimB and FimE, which mediate an 'on' or 'off' configuration of the switch, respectively. In this study, we have investigated how these proteins control the switch by means of fim-lac fusions on low-copy-number plasmids. It was found, by in trans and cis complementation, that the ratio of fimB to fimE and the total concentration of the gene products determine the configuration of the switch as well as the frequency of phase switching. It was also shown that transcription occurs from the promoter located at the phase switch when this is in the 'off' configuration. This suggests a regulatory mechanism, since the resulting transcript would be anti-sense to the fimE transcript. PMID- 2571916 TI - The proposed role of neurotransmitter receptors in the pathophysiology of human myoclonic disorders. AB - The hypothesis that central neurotransmitter receptor abnormalities are the basis of human myoclonic disorders is novel. Receptor abnormalities in any of several different neurotransmitter systems implicated in myoclonus may be genetic or the consequence of various brain injuries. These abnormalities might define pharmacologic subgroups of possible relevance to the clinical, neurophysiologic, and pathologic heterogeneity of myoclonus. Receptor abnormalities may be the primary pathophysiologic defect, involving the surface recognition site or effector-transducer mechanisms in the defect, involving the surface recognition site or effector-transducer mechanisms in the post-synaptic cell. Alternatively, changes in receptor density or affinity may be adaptive (recovery of function) or maladaptive (supersensitivity, subsensitivity, dysregulation). Drug treatments, then, could be targeted at the receptor changes, reversing abnormalities and enhancing compensatory mechanisms. Current therapy may inadvertently have such an effect. Polytherapy may be justified to target more than one component of the "pharmacologic receptor". Identification of receptor abnormalities in human post mortem brain may have diagnostic and therapeutic significance. New advances in the pharmacologic selectivity of receptor agonists and antagonists and in the measurement of receptors should be applied to the problem of myoclonus. PMID- 2571917 TI - [Beta receptor blockers. Principles for drug selection for rational therapy]. PMID- 2571918 TI - Hepatoblastoma and polyposis coli (familial adenomatous polyposis) PMID- 2571920 TI - AIDS: The good news and the bad. PMID- 2571919 TI - [HIV infection in stage IV C-2 (CDC). Increase in the p24 antigen value before critical decrease in CD4+ lymphocytes]. AB - The Center for Disease Control proposed a classification system of HIV-infection including a symptomatic stage IV C-2 with secondary infections like thrush, oral hairy leukoplakia and herpes zoster. Because the prognostic value of these symptoms for the development of AIDS has been proven and the CDC-classification does not include any immunological parameters we analyzed the numbers of CD4+ lymphocytes and the frequency of HIV-antigen in 65 HIV-infected individuals. When entering stage IV C-2, the incidence of HIV-antigenemia was as high as in full blown AIDS (53% and 60%, respectively). However, we observed no decrease of CD4+ lymphocytes as compared to earlier CDC-stages.--We conclude that in stage IV C-2 the increase of viral protein production proceeds independently from CD4+-status. PMID- 2571921 TI - [Immunohistochemical localization of nerve terminals in the human gingiva. Use of a new neuronal marker]. AB - The usefulness of classical techniques for sensory nerve ending detection is compromised by intrinsic limitations. The development of immunohistochemical methods has recently produced new information on the innervation pattern in different human tissues. This study presents the results obtained by use of a new antiserum, raised against a new marker of central and peripheral neurons (PGP 9.5), which selectively labels nerve fibers and neuroendocrine (APUD) cells also in human gingiva. The possible application of this method for further researches is also briefly discussed. PMID- 2571922 TI - Initial clinical management of the HIV-infected patient. AB - For the primary care physician, treatment of HIV-infected patients often centers on the stage of illness at which the patient first seeks care. Disease staging, which uses symptoms in its classification scheme, thus leads to evaluation and selection of appropriate therapy. This is the second article in a series from the Washington University AIDS Clinical Trials Unit. PMID- 2571923 TI - The mouse carbonic anhydrase I gene contains two tissue-specific promoters. AB - We report the isolation and characterization of the mouse carbonic anhydrase I (CAI) gene. Direct RNA sequence analysis of the 5' nontranslated regions of CAI mRNA from mouse colon and mouse erythroleukemia cells demonstrated tissue specificity in the lengths and sequences of CAI transcripts. Analysis of several mouse CAI genomic clones showed that the transcripts arose from a single CAI gene with two tissue-specific promoters and eight exons. CAI transcripts in the colon were found to initiate just upstream of the erythroid exon 2 of the CAI gene region sequence. Erythroid transcripts originated from a novel promoter upstream of exon 1, which was located more than 10 but less than 250 kilobases upstream of exon 2. Erythroid exon 1 contained only a nontranslated sequence, which was spliced to exon 2 via a cryptic splice acceptor site located in the region that encoded the colon mRNA 5' nontranslated sequence. The remaining exon-intron junctions were conserved in comparison with those of the CAII and CAIII genes. PMID- 2571925 TI - Microinjected Xenopus oocytes secrete mature, biologically active parathyroid hormone. AB - Xenopus oocytes have been shown to faithfully translate, process, and secrete a number of secretory proteins after the injection of heterologous mRNAs. The oocyte has the capacity to perform a variety of posttranslational protein modifications but has been reported to be incapable of carrying out certain two step cleavages which proceed via propeptide intermediates. We examined the ability of the oocyte to process preproPTH after the injection of parathyroid mRNA. Microinjected oocytes secreted material which could be detected in a sensitive cytochemical bioassay for PTH. This activity paralleled that of the PTH standard in the assay and was entirely eliminated by a competitive inhibitor of PTH binding, by preincubation with an anti-PTH antiserum, and by coinjecting oocytes with an oligonucleotide mixture complementary to PTH sequences. Immunoprecipitable proPTH and PTH were present in oocyte homogenates, but oocyte conditioned medium contained only mature PTH(1-84). We conclude that the Xenopus oocyte is capable of accurately processing preproPTH to the mature secretory form of the peptide. PMID- 2571924 TI - Multiple genes coding for precursors of rhodotorucine A, a farnesyl peptide mating pheromone of the basidiomycetous yeast Rhodosporidium toruloides. AB - Haploid cells of mating type A of the basidiomycetous yeast Rhodosporidium toruloides secrete a mating pheromone, rhodotorucine A, which is an undecapeptide containing S-farnesyl cysteine at its carboxy terminus. To analyze the processing and secretion pathway of rhodotorucine A, we isolated both genomic and complementary DNAs encoding the peptide moiety. We identified three distinct genes, RHA1, RHA2, and RHA3, encoding four, five, and three copies of the pheromone peptide, respectively. Complementary DNA clones were classified into two types. One type was homologous to RHA1, and the other type was homologous to RHA2. Transcription start sites were identified by primer extension and S1 nuclease protection, from which the site of the initiator methionine was verified. A primary precursor of rhodotorucine A was detected as a 7-kilodalton protein by immunoprecipitation of in vitro translation products. On the basis of these results, we propose similar three-precursor structures of rhodotorucine A, each containing the amino-terminal peptide sequence Met-Val-Ala. The precursors contain three, four, or five tandem repeats of the pheromone peptide, each separated by a spacer peptide, Thr-Val-Ser(Ala)-Lys, and each precursor has the carboxy-terminal sequence Thr-Val-Ala. This structure suggests that primary precursors of rhodotorucine A do not contain canonical signal sequences. PMID- 2571926 TI - [Complications after preventive mumps vaccination in West Germany (including multiple preventive vaccinations)]. AB - Since the introduction of mumps vaccination in the Federal Republic of Germany 34 untoward vaccinal reactions were collected. In 2 cases of aseptic meningitis after mumps immunisation mumps virus was isolated from the CSF. The final investigation showed mumps virus (wild type) in one case; in the other a mumps vaccine-like strain (Jeryl-Lynn) was identified. Both patients recovered completely. These complications do not outweigh the enormous benefits of mumps immunisation. PMID- 2571927 TI - Modulation of major histocompatibility complex antigens and inhibition of proliferative activity of YAC lymphoma cells by a natural killer lysis resistance inducing factor (NK-LRIF). AB - Concanavalin-A-activated rat spleen cells secrete a natural killer lysis resistance-inducing factor (NK-LRIF) distinct from interleukin-2 and interferon, which induces resistance to NK-cell-mediated lysis in YAC tumor cells. In order for NK-LRIF to have an effect on YAC cells, several hours of incubation is required. When NK-LRIF-treated YAC cells are washed and cultured in the absence of NK-LRIF, normal NK susceptibility is regained. YAC cells treated with NK-LRIF show a significant decrease in the rate of proliferation as judged by changes in cell numbers and rate of thymidine incorporation. Cell cycle studies indicate that the proportion of G0/G1 phase cells increases in YAC preparations treated with NK-LRIF. Major histocompatibility complex (MHC) class I antigen expression is markedly enhanced on YAC cells incubated with NK-LRIF but the expression of MHC class II antigens and Thy-1 antigen remains unchanged. No effect of NK-LRIF treatment on the capacity of YAC cells to bind effector spleen cells could be demonstrated. PMID- 2571928 TI - New directions in the management of Zollinger-Ellison syndrome: a case study and review. PMID- 2571929 TI - A genomic change associated with the development of resistance to hycanthone in Schistosoma mansoni. AB - Ribosomal gene probes were used to investigate the genetic basis of drug resistance in schistosomes in a model where resistance to the anthelmintic hycanthone (HC) is generated by exposing immature worms to the drug. Two strains of Schistosoma mansoni, JHU and NMRI, were used. Drug resistance could be produced in the JHU strain by treatment with HC, but was also found to occur spontaneously. In contrast, it was not possible to detect or produce resistance to HC in the NMRI strain. A genomic alteration accompanied the development of resistance. The change was evidence by the occurrence of restriction fragment length polymorphisms (RFLPs) when Southern blots of genomic DNA from HC-resistant worms were hybridized with the ribosomal probe pSM389, which contains part of the small rRNA gene plus non-transcribed spacer (NTS) sequence. The most reliable marker of HC-resistance was a 3.6-kb BamHI fragment which was present and heritable in 7 drug-resistant lines derived from the JHU strain but absent from the parent JHU population and from NMRI parasites. The universal absence of the 3.6-kb RFLP in HC-sensitive individuals and its presence in the drug-resistant progeny suggest that resistance results from an induced change in the population rather than from selection of HC-resistant parasites. The rRNA gene sequence responsible for detecting the 3.6-kb RFLP appears to be localized either to the NTS or to the 5' end of the small rRNA gene, since hybridization to a probe containing sequence from the rRNA gene contiguous and downstream from the insert of pSM389 failed to reveal the RFLP. These results show that the development of resistance to HC is accompanied by a genomic rearrangement. PMID- 2571930 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 42-1989. A 64-year-old woman with a liver abscess, Clostridium perfringens sepsis, progressive sensorimotor neuropathy, and abnormal serum proteins. PMID- 2571932 TI - The beneficence of neonatal hematopoiesis. PMID- 2571931 TI - Hematopoietic reconstitution in a patient with Fanconi's anemia by means of umbilical-cord blood from an HLA-identical sibling. PMID- 2571933 TI - Amyloid A4 protein and its precursor in Down's syndrome and Alzheimer's disease. PMID- 2571934 TI - The caudal gene product is a direct activator of fushi tarazu transcription during Drosophila embryogenesis. AB - A drosophila pair-rule segmentation gene, fushi tarazu (ftz), encodes a protein which is expressed in a characteristic seven-stripe pattern. The promoter sequences that are sufficient for generating this spatially restricted pattern of expression are located within 669 base pairs upstream of the transcription start site. Multiple transcriptional activators and repressors interact with this 'zebra-stripe' promoter unit to bring about the positional specificity of ftz transcription. Here we report that the homoeodomain-containing protein encoded by caudal (cad) is one such regulator. The cad gene product can increase the level of ftz transcription in the posterior half of the embryo by interacting with multiple copies of a TTTATG consensus sequence located in the zebra-stripe unit. This result demonstrates one pathway by which the product of a maternally expressed segmentation gene, expressed in an antero-posterior concentration gradient, can directly regulate the expression of a pair-rule gene. PMID- 2571935 TI - Vertebrate development: genes and segmentation. PMID- 2571936 TI - Segmental expression of Hox-2 homoeobox-containing genes in the developing mouse hindbrain. AB - The vertebrate hindbrain develops in a segmental pattern, with distinctive groups of neurons originating from different segments. We report here that members of the Hox-2 cluster of murine homoeobox genes are expressed in segment-specific patterns in the developing hindbrain, with successive genes having boundaries at two-segment intervals. These data indicate that Hox genes specify segment phenotype, a role analogous to that of their Drosophila homologues. PMID- 2571937 TI - The Oct-1 homoeodomain directs formation of a multiprotein-DNA complex with the HSV transactivator VP16. AB - The herpes simplex virus transactivator VP16 participates in the formation of a multiprotein-DNA complex with the ubiquitous octamer-motif-binding factor Oct-1. Complex formation is dependent on specific amino acids in the Oct-1 homoeodomain which are in positions analogous to positive control mutations in helix 2 of the lambda phage repressor helix-turn-helix motif, indicating that this structure is an ancient target for protein-protein interactions mediating transcriptional control. PMID- 2571938 TI - Positive selection of CD4+ T cells mediated by MHC class II-bearing stromal cell in the thymic cortex. AB - T lymphocytes differentiate in the thymus, where functionally immature, CD4+CD8+ (double positive) thymocytes develop into functionally mature CD4+ helper cells and CD8+ cytotoxic (single positive) T cells. The thymus is the site where self reactive T cells are negatively selected (clonally deleted) and where T cells with the capacity to recognize foreign antigens in association with self-proteins encoded by the major histocompatibility complex (MHC) are positively selected. The net result of these developmental pathways is a T-cell repertoire that is both self-tolerant and self-restricted. One unresolved issue is the identity of the thymic stromal cells that mediate the negative and positive selection of the T-cell repertoire. Previous work has pointed to a bone-marrow-derived macrophage or dendritic cell as the inducer of tolerance, whereas a radiation-resistant, deoxyguanosine-resistant thymic cell seems to mediate the positive selection of self-MHC restricted T cells. Thymic stromal cells in the cortex interact with the T-cell antigen receptor on thymocytes. Using several strains of transgenic mice that express the class II MHC molecule I-E in specific regions of the thymus, we show directly that the positive selection of T cells is mediated by an I-E bearing cell in the thymic cortex. PMID- 2571939 TI - Arachidonic acid induces a long-term activity-dependent enhancement of synaptic transmission in the hippocampus. AB - Long-term potentiation (LTP) is a widely studied model of the synaptic basis of information storage in the mammalian brain. The induction of LTP is triggered by the postsynaptic entry of calcium through the channel associated with the N methyl-D-aspartate (NMDA) receptor, whereas its maintenance is mediated, at least in part, by presynaptic mechanisms. To explain how postsynaptic events can lead to an increase in transmitter release, we have postulated the existence of a retrograde messenger to carry information from the postsynaptic side of the synapse to recently active presynaptic terminals. Candidates for a retrograde messenger include arachidonic acid or one of its lipoxygenase metabolites. Here we report that weak activation of the perforant path, when given in the presence of arachidonic acid, leads to a slow-onset persistent increase in synaptic efficacy both in vivo and in vitro. The activity-dependent potentiation thus produced is accompanied by an increase in the release of glutamate, and is non additive with tetanus-induced LTP. These observations indicate a role for arachidonic acid as a retrograde messenger in the later, but not the initial, stages of LTP. PMID- 2571941 TI - Characterization of a homologue of bithorax-complex genes in the leech Hirudo medicinalis. AB - We report the isolation and characterization of the Hirudo medicinalis homoeobox gene Lox2. Sequence analysis shows that it contains a region that has homology to Drosophila and vertebrate homoeodomains of the Antennapedia class. In addition, Lox2 shares homology with sequences in the bithorax complex Ultra-bithorax (Ubx) and abdominal A (abdA) genes in a region adjacent to the C-terminus of the homoeodomain. Whole mount in situ hybridization of embryos of various ages demonstrates that during early development this gene has temporally and spatially restricted patterns of expression that resemble those of the homoeotic genes of the Drosophila bithorax complex and of many vertebrate homoeobox genes. The largest accumulation of transcripts was seen in the posterior two-thirds of the developing leech central nervous system in 7-14-day-old embryos. Adult leeches also express Lox2. We propose that in Hirudo, Lox2 represents the ancestral gene of the Ubx and abdA genes of the bithorax complex of Drosophila. PMID- 2571940 TI - Selective development of CD4+ T cells in transgenic mice expressing a class II MHC-restricted antigen receptor. AB - T lymphocytes are predisposed to recognition of foreign protein fragments bound to cell-surface molecules encoded by the major histocompatibility complex (MHC). There is now compelling evidence that this specificity is a consequence of a selection process operating on developing T lymphocytes in the thymus. As a result of this positive selection, thymocytes that express antigen receptors with a threshold affinity for self MHC-encoded glycoproteins preferentially emigrate from the thymus and seed peripheral lymphoid organs. The specificity for both foreign antigen and MHC molecules is imparted by the alpha and beta chains of the T-cell antigen receptor (TCR). Two other T-cell surface proteins, CD4 and CD8, which bind non-polymorphic regions of class II and class I MHC molecules respectively, are also involved in these recognition events and play an integral role in thymic selection. In order to elucidate the developmental pathways of class II MHC-restricted T cells in relation to these essential accessory molecules, we have produced TCR-transgenic mice expressing a receptor specific for a fragment of pigeon cytochrome c and the Ek (class II MHC) molecule. The transgenic TCR is expressed on virtually all T cells in mice expressing Ek. The thymuses of these mice contain an abnormally high percentage of mature CD4+CD8- cells. In addition, the peripheral T-cell population is almost exclusively CD4+, demonstrating that the MHC specificity of the TCR determines the phenotype of T cells during selection in the thymus. PMID- 2571942 TI - Suppression of PAF-induced bronchoconstriction in the guinea-pig by oxatomide: mechanism of action. AB - Oxatomide potently (ED50 0.9 mg/kg orally, -2 h) attenuates the reduction of pulmonary tidal volume elicited by PAF (250 ng/kg i.v.) in anaesthetized, ventilated and propranolol-treated guinea-pigs. The increase of the pulmonary inflation pressure elicited by PAF (40 ng/kg i.v.) in such animals, ventilated at a fixed tidal volume, is also significantly reduced by the compound, but substantially higher doses (5 mg/kg i.v., -15 min) are required. The potency of oxatomide in the latter respect (50.4% reduction) is equivalent to that of ketotifen at 5 mg/kg i.v. (55% reduction). In spontaneously breathing, anaesthetized guinea-pigs, oxatomide (5 mg/kg i.p., -1 h) significantly reduces the increase in pulmonary resistance, but not the reduction in dynamic compliance, elicited by PAF (30, 60, 90 ng/kg i.v.), suggesting a pharmacological interference mainly with PAF-induced processes in the larger airways. Changes in arterial blood pressure, haemoconcentration, thrombocytopenia and leukopenia induced by PAF in vivo, contraction of guinea-pig lung parenchymal strips, production of superoxide anion by alveolar macrophages, aggregation and release of ATP by platelets challenged with PAF in vitro are not affected by the compound. These observations suggest that oxatomide attenuates the PAF-induced pulmonary reactions by inhibiting the release and/or the effect of allergic mediators elicited by the phospholipid rather than by a direct antagonism at the PAF receptors. PMID- 2571943 TI - Kinetic properties of the in vivo accumulation of 3H-(-)-N-n-propylnorapomorphine in mouse brain. AB - (1) The influence of various dopamine (DA) receptor agonists and antagonists on the kinetic properties of the specific binding of 3H(-)-N-n-propylnorapomorphine (NPA) in the mouse striatum in vivo was studied. The specific binding of 3H-NPA, defined as the difference between the radioactivity in the striatum and cerebellum, was completely antagonized by the selective D-2 receptor antagonist raclopride but not by the selective D-1 antagonist SCH 23390, showing that the binding occurs exclusively to the D-2 receptors. (2) The selective D-2 receptor agonists pergolide and quinpirole inhibited the 3H-NPA binding biphasically at low doses, indicating that these DA receptor agonists have high affinities for a subfraction (10 to 30%) of the NPA binding sites. (3) Increasing the synaptic DA concentration by DA release [(+)-amphetamine] or uptake blockade (amfonelic acid and methylphenidate) inhibited the 3H-NPA binding in a competitive manner (unchanged Bmax, increased KD). Depletion of the DA in the synapses by gamma butyrolactone or reserpine decreased the apparent KD value. (4) The possibility of estimating changes in the synaptic DA concentration from changes in the apparent KD is discussed. According to the results obtained, the normal concentration of DA in the synaptic cleft in mouse striatum in vivo is about 40 nmol/l and this concentration is increased 2 to 3 times by (+)-amphetamine and amfonelic acid in doses which evoke hyperactivity and stereotypic behaviour. PMID- 2571944 TI - Occurrence of yawning and decrease of prolactin levels via stimulation of dopamine D2-receptors after administration of SND 919 in rats. AB - SND 919 [S)-2-amino-4,5,6,7-tetrahydro-6-propylamino-benzothiazole) is expected to have a potent and selective dopamine D2-receptor agonistic activity. From this information, the present study was performed to investigate effects of SND 919 on yawning behavior and prolactin secretion in rats. Subcutaneous injections of SND 919 (25-500 micrograms/kg, s.c.) elicited yawning responses. Its dose-response curve was bell-shaped with maximal effects at a dose of 100 micrograms/kg. Yawning behavior was also evoked by the putative dopamine autoreceptor agonists, talipexole (6-allyl-2-amino-5,6,7,8-tetrahydro-4H-thiazolo [4,5-d]azepine) (B-HT 920) (5-100 micrograms/kg, s.c.) and (+)-3-PPP ((+)-3-(3-hydroxyphenyl)-N-n propylpiperidine) (5-15 mg/kg, s.c.). The yawning induced by SND 919 (100 micrograms/kg, s.c.) as well as talipexole (25 micrograms/kg, s.c.) was inhibited by pretreatment with dopamine D2-receptor antagonists such as spiperone (0.5 mg/kg, i.p.) and YM-09151-2 (cis-N-(1-benzyl-2-methylpyrrolidin-3-yl)-5-chloro-2 methoxy-4-met hylamino- benzamide) (0.1 mg/kg, i.p.), or the muscarinic receptor antagonist, scopolamine (0.5 mg/kg, i.p.). However, the yawning was not affected by the dopamine D1-receptor antagonist, SCH 23390 (R(+)-8-chloro-2,3,4,5 tetrahydro-3-methyl-5-phenyl-1H-3-benzazepine-7-o l) (0.5 mg/kg, i.p.). Stereotypy such as licking and biting was not observed following the administration of SND 919, talipexole and (+)-3-PPP. Administration of SND 919, talipexole or (+)-3-PPP in respective yawn-inducing doses caused a reduction in both the basal prolactin levels and the alpha-methyl-p-tyrosine-induced hyperprolactinemia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571945 TI - Involvement of central beta-adrenoceptors in the regulation of yawning responses. AB - A behavioral study was performed in an attempt to understand the role of central beta-adrenoceptors in yawning in rats. Yawning was evoked by apomorphine and piribedil, mixed dopamine D1/D2-receptor agonists, but not by SK&F 38393 [1 phenyl-2,3,4,5-tetrahydro-(1H)-3-benzazepine-7,8-diol], a dopamine D1-receptor agonist. The apomorphine-induced yawning was increased by pindolol, propranolol, indenolol, alprenolol and bukumolol which block the central beta-adrenoceptors, but not by the peripheral beta-adrenoceptor antagonists, carteolol and atenolol. These beta-adrenoceptor antagonists given alone did not elicit yawning. Conversely, the yawning was inhibited by salbutamol, a beta-adrenoceptor agonist, without being affected by prazosin, an alpha-adrenoceptor antagonist. The combined administration of SK&F 38393 and the beta-adrenoceptor antagonists did not induce yawning. The yawning elicited by either apomorphine or piribedil in combination with pindolol was suppressed by spiperone and YM-09151-2 [cis-N-(1 benzyl-2-methyl-pyrrolidin-3-yl)-5- chloro-2-methoxy-4-methylamino-benzamide], dopamine D2-receptor antagonists, and scopolamine, a muscarinic receptor antagonist, but not by SCH 23390 [R(+)-8-chloro-2,3,4,5-tetrahydro-3-methyl-5 phenyl-1H-3- benzazepine-7-ol], a dopamine D1-receptor antagonist. Physostigmine or pilocarpine induced yawning, which was also enhanced by pindolol and propranolol. This enhanced yawning was inhibited by scopolamine, but not by spiperone, YM-09151-2 and SCH 23390.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2571948 TI - [Molecular genetics of Duchenne's disease]. PMID- 2571946 TI - Phentolamine blocks presynaptic serotonin autoreceptors in rabbit and rat brain cortex. AB - Possible antagonist effects of phentolamine at presynaptic serotonin autoreceptors were studied in slices of the occipito-parietal cortices of the rabbit and the rat. The slices were preincubated with 3H-serotonin and then superfused and stimulated electrically with single pulses or pulse trains. Nitroquipazine 1 mumol/l, a compound that inhibits the high affinity neuronal uptake of serotonin, was present in the superfusion medium in all one pulse experiments as well as in experiments in which the effect of unlabelled serotonin was examined. In rabbit cortical slices, unlabelled serotonin reduced the single pulse-evoked overflow of tritium. Its concentration-response curve was not changed by the selective alpha 2-adrenoceptor antagonist idazoxan 1 mumol/l but was shifted to the right by phentolamine 1 and 10 mumol/l. Phentolamine 10 mumol/l also shifted to the right the concentration-inhibition curve of the selective 5-HT1-receptor agonist 5-carboxamidotryptamine. When the slices were stimulated by trains of 30 pulses at 3 Hz, phentolamine 1 and 10 mumol/l but not 0.1 mumol/l increased the evoked overflow of tritium, the maximal increase amounting to 178%; its effect was enhanced in the presence of nitroquipazine 1 mumol/l plus idazoxan 10 mumol/l (a drug combination that, when given alone, slightly increased the evoked overflow of tritium). The serotonin receptor antagonist metitepin at concentrations of 0.01-1 mumol/l also increased the overflow of tritium elicited by 30 pulses/3 Hz, the maximal increase amounting to 280%; its effect was potentiated in the presence of nitroquipazine 1 mumol/l plus idazoxan 10 mumol/l but was abolished or almost abolished in the presence of nitroquipazine 1 mumol/l plus phentolamine 10 mumol/l (a drug combination that, given alone, greatly increased the evoked overflow of tritium). When slices were stimulated by trains of 360 pulses at 3 Hz, there was no apparent antagonism of phentolamine 10 mumol/l against the inhibitory effect of unlabelled serotonin. In rat brain cortex slices, unlabelled serotonin reduced the overflow of tritium elicited by 4 pulses delivered at 100 Hz. Again, phentolamine 10 mumol/l shifted the concentration-response curve to the right. It is concluded that phentolamine blocks presynaptic serotonin autoreceptors in rabbit and rat brain cortex with pA2 values of 6.44 and 5.95, respectively. Previous failures to detect the antagonistic effect against exogenous agonists were probably due to stimulation conditions that led to marked endogenous autoinhibition of serotonin release.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2571947 TI - Cerebral-activating (EEG) properties of two inverse agonists and of an antagonist at the benzodiazepine receptor in the rat. AB - In order to assess the effects of inverse benzodiazepine agonists and antagonists on brain function, computerized EEG (CEEG) analysis was performed in rats following the i.p. administration of SR 95195 (7-phenyl-3-methyl-1,2,4 triazolo [4,3-b]pyridazine) and CGS 8216 (2-phenylpyrazolo-[4,3c]-quinoline-3-[5H]-one) two benzodiazepine receptor inverse agonists (BRIAGs) and of flumazepil (Ro 15 1788), a benzodiazepine receptor antagonist (BRANT). The EEG effects of SR 95195 (3, 10, 30 and 60 mg/kg), CGS 8216 (10 and 30 mg/kg) and flumazepil (3, 10, 30 and 60 mg/kg) were compared to those of the psychostimulant drugs DL-amphetamine (0.1, 0.3 and 1 mg/kg), and caffeine (10 and 30 mg/kg) and those of aniracetam (100 and 300 mg/kg), a nootropic pyrrolidone derivative. The CEEG profiles of SR 95195, CGS 8216 and flumazepil were mainly characterized by a power increase in the 20-32 Hz frequency range and by a power reduction in the 8-16 Hz range. These effects were quite similar to those of the psychostimulants DL-amphetamine and caffeine as well as to those of the nootropic aniracetam. Other psychotropic drugs with CNS-depressant properties, namely diazepam (10 mg/kg p.o.), pentobarbital (30 mg/kg p.o.), chlorpromazine (10 mg/kg i.p.) and imipramine (10 mg/kg i.p.) induced quite different EEG power modifications. These results show that BRIAGs and BRANTs possess a marked intrinsic activity at the central level and suggest that this activity is CNS-activating in nature. PMID- 2571949 TI - [The effect of immunoscintigraphy with monoclonal antibodies on assays of hormones and tumor markers. This is not the end of the matter!]. AB - The use of monoclonal antibodies in medicine for in-vivo diagnostic methods and for therapeutic purposes will increase in the future. Although monoclonal antibodies possess a high specificity, the animal origin of these antibodies remains a problem. Repeated administration of animal monoclonal antibodies (in vivo) may induce the formation of human antibodies against these monoclonal antibodies. Because animal monoclonal antibodies are also used in laboratory assays (in vitro), the presence of human antibodies against these animal monoclonal antibodies may cause spuriously elevated or depressed results of these assays. The clinician should be alert to this possibility. A case history is presented to demonstrate the problem. PMID- 2571951 TI - [Exogenous factors and schizophrenia]. PMID- 2571950 TI - [Sulfasalazine allergy: fever, skin rash, hepatitis and T-lymphocytes]. AB - We describe a case of severe sulfasalazine allergy. Exacerbation of the symptoms occurred after unintentional rechallenge with co-trimoxazole, indicating that the reaction was triggered by the sulphonamide component. The clinical picture consisted of generalised adenopathy, hepatitis, high fever and a maculopapular skin rash. A bone marrow biopsy and skin biopsy both showed noncaseating granulomas. The white blood cell count rose to 90.10(9)/l with 40% atypical lymphocytes (plasmacytoid). They were identified by flow cytometry as activated T lymphocytes. PMID- 2571952 TI - [A drowsy child: benzodiazepine poisoning?]. AB - In The Netherlands accidental intoxications in children due to benzodiazepines are regularly encountered. In 1987 of 1630 requests for information at the National Poison Control Centre about probable benzodiazepine intoxications 144 (8.8%) concerned children 0-12 years of age. The symptoms of this type of intoxication are non-specific and if the physician does not think of benzodiazepine intoxication extensive diagnostic procedures may be performed. If children show symptoms e.g. unconsciousness, ataxia and hypotonia, the physician should always think of benzodiazepine intoxication and try to confirm or to exclude this possibility by toxicological analysis. We discuss ways and means of the diagnosis and how to avoid pitfalls on the way. PMID- 2571953 TI - Sexual and developmental differences in peptides regulating growth hormone secretion in the rat. AB - Sex differences in the hypothalamic control of growth hormone (GH) secretion were investigated by measuring rat GH-releasing factor (rGRF) and somatostatin in male and female rats. Rat GRF-like immunoreactivity (rGRF-IR) was higher in the median eminence and hypothalamic tissue outside of the median eminence of adult (90-day old) male compared to female rats. A similar pattern of rGRF-IR content was found in the median eminence of 35-day-old rats. This sex difference developed between days 25 and 35 of age, during which time serum concentrations of insulin-like growth factor (IGF-1) and body weight increased in both sexes. To a lesser extent, the content of somatostatin-like immunoreactivity (SLI) was higher in the median eminence of adult female rats compared to male rats. Whole hypothalamic rGRF-IR and SLI contents were influenced only moderately by adult gonadectomy or gonadal steroid treatments. For example, estrogen increased rGRF-IR content in castrated rats, but orchidectomy alone or orchidectomy followed by testosterone did not influence rGRF-IR content. Additionally, whole hypothalamic SLI content was unaffected by orchidectomy or orchidectomy followed by testosterone or estrogen. One month after ovariectomy, rGRF-IR and SLI in whole hypothalamic fragments were similar to their respective contents in gonad-intact males. However, ovariectomy followed by estrogen or testosterone did not restore rGRF-IR content and partially restored SLI content to levels seen in gonad-intact females. PMID- 2571954 TI - Effect of water deprivation and salt loading on atrial natriuretic peptide stimulated guanylate cyclase activity in the rat subfornical organ. AB - The effect of water deprivation and salt loading on rat atrial natriuretic peptide (99-126) (rANP)-stimulated guanylate cyclase activity was investigated in the rat subfornical organ. rANP stimulated the formation of cGMP in rat subfornical organ crude homogenates in a dose- and time-dependent manner. An elevated responsiveness to rANP-induced cGMP production was observed in the subfornical organ after 4 days of water deprivation; on the contrary, after salt loading the response to the cGMP-generating effects of ANP were less pronounced than those in the corresponding control tissue. Our results suggest that cGMP mediates at least some of the central actions of rANP through the activation of specific receptors in localized target sites, and they provide evidence suggesting that the guanylate cyclase-coupled ANP-binding sites are susceptible to the regulatory mechanism described in the rat subfornical organ. PMID- 2571955 TI - Corticotropin-releasing factor neurons innervate dopamine neurons in the periventricular hypothalamus of juvenile macaques. Synaptic evidence for a possible companion neurotransmitter. AB - Corticotropin-releasing factor (CRF) and dopamine (DA) are important integrators of the endocrine and autonomic response to stress. CRF neurons in the anterior portions of the periventricular nucleus (PV) and parvocellular paraventricular nucleus (pvPVN) occur close to A14 DA neurons in these same locations. Since CRF has been shown to act as an excitatory neurotransmitter, possible CRF interactions with the DA system were investigated using double-label immunocytochemistry. Coronal vibratome sections through the PV and pvPVN were obtained from colchicine-treated and nontreated juvenile female cynomolgus macaques. They were sequentially immunostained for tyrosine hydroxylase (TH) (to identify DA neurons) with PAP and DAB, and for CRF using 15 nm colloidal gold. By light microscopy, areas of coincidence of TH- and CRF-immunoreactive cell bodies in the PV and pvPVN were obvious, but double-stained elements were not observed. By electron microscopy, asymmetrical synapses frequently occurred between CRF axons and TH dendrites or somata. Symmetrical axosomatic synapses sometimes appeared adjacent to these CRF/TH synapses, while symmetrical axoaxonic synapses were rare. We conclude that CRF neuronal efferents synaptically activate A14 DA neurons in the primate PV and pvPVN. Parallel CRF/DA symmetrical synapses also suggest coexistence of a companion transmitter within some of these same CRF neurons. Our own previous work and recent independent studies indicate that this transmitter is probably GABA. Thus the CRF neuronal system, which is known to alter secretion of several pituitary hormones, may also act through hypothalamic periventricular DA neurons to mediate other responses to stress. PMID- 2571956 TI - Immunocytochemical localization of the intermediate filament protein peripherin in adult mouse adrenal chromaffin cells in culture. AB - Peripherin is the main intermediate filament protein in sympathetic neurons. Immunoreactivity to peripherin was studied in mouse adrenal chromaffin cells after 6 days in culture, and compared to immunoreactivity to tyrosine hydroxylase used as a general marker of chromaffin cells in culture. Most of the cells immunoreactive to tyrosine hydroxylase were rounded, with a glandular phenotype and a few of them had processes. The cells reactive to peripherin only constituted a small proportion of the chromaffin cells (2%), and most of them sent out processes. However, not all the cells with processes were reactive for peripherin. These results did not change in the presence of nerve growth factor. The discussion focuses on the significance of the sub-population of cells reactive to peripherin. We suggest that these cells resemble the small granule chromaffin cells, regarded as an intermediate cell type between glandular cells and neurons. The cells that expressed peripherin here are compared to those selected to form the PC12 clone. The presence of peripherin in only a few of the cells sending out neurite-like processes is discussed in relation to the expression of other neurofilament proteins in developing cells and to the influence of non-chromaffin cells. PMID- 2571957 TI - [The hemodynamic effects on the portal system of levomoprolol and propranolol in rats with an enhanced adrenergic tonus]. PMID- 2571960 TI - AIDS therapy. PMID- 2571959 TI - Complications arising from the use of ophthalmologic medications in an intensive care unit patient. PMID- 2571958 TI - Interaction of noradrenergic and cholinergic systems in regulation of ocular dominance plasticity. AB - We studied interactions among the noradrenergic (NA) and the muscarinic cholinergic (ACh) systems in the regulation of ocular dominance plasticity in kitten visual cortex. The cortex was bilaterally infused with 6-hydroxydopamine (6-OHDA) for a week. Upon termination of the 6-OHDA infusion, one hemisphere was infused with a muscarinic ACh agonist, bethanechol, through the same, chronically implanted cannula for the second week together with monocular lid suture. The other hemisphere received an infusion of the vehicle solution alone. (1) Only in the hemisphere infused with bethanechol at relatively high concentrations did we obtain a clear shift in ocular dominance. We also found that the effect of bethanechol was concentration-dependent. (2) By comparing necessary concentrations of bethanechol and NA for the respective maximal effects, we noted that the former was at least 100-fold less effective than the latter in restoring the plasticity. (3) The cortical infusion of bethanechol did not restore the plasticity to the propranolol-pretreated cortex; the ocular dominance distribution remained virtually unchanged. This result was interpreted as suggesting that functioning beta-adrenoreceptors are needed for the cortical effect of activating the muscarinic ACh receptors to become detectable. (4) The expected shift in ocular dominance following monocular deprivation was partially suppressed, when highly concentrated scopolamine, a muscarinic ACh antagonist, was used, indicating that the involvement of the ACh system in this matter was indirect. The concentration of scopolamine needed for the half-maximum effect was 172-fold higher than that of propranolol. We thus conclude that the involvement of the muscarinic ACh system in ocular dominance plasticity is secondary to that of the NA-beta-adrenoreceptor system. PMID- 2571962 TI - Management of foot problems. PMID- 2571961 TI - Xerostomia. Part II: Relationship to nonoral symptoms, drugs, and diseases. AB - Five hundred twenty-nine adult outpatients were studied to determine the relationship of xerostomia to other oral symptoms and salivary flow (reported in part I) and to nonoral symptoms, drugs, and select diseases (reported here in Part II). It was observed that dry throat, blurred vision, dry eyes, dry skin, and vaginal itching and fungal infections are prominently associated with oral dryness. These nonoral symptoms were positively correlated with the oral symptoms cited in part I of this study and were inversely related to the flow of resting, but not stimulated, whole saliva. Several classes of drugs were associated with dry mouth. In addition, diabetes mellitus and hypertension were significantly associated with it. Approximately half of the diabetic and hypertensive patients complained of dry mouth. Although a majority of them were taking medications, the association between xerostomia and these diseases cannot be completely attributed to drugs, since many of these patients did not take any xerogenic medicaments. The data show that xerostomia and several other oral symptoms are valid indicators of salivary gland hypofunction. They suggest, moreover, that select nonoral symptoms are an indicator of generalized xerosis. PMID- 2571963 TI - Foot and ankle injuries in classical ballet dancers. AB - The pathomechanics of dance injury are explained. Specific injuries are included, such as fractures, ankle sprains, anterior impingement syndrome, posterior impingement syndrome, flexor hallicus longus tendinitis, Achilles tendinitis, and stress fractures. PMID- 2571964 TI - [Adaptation reactions of the body in fractures and fracture-dislocations of the bones of the foot]. AB - Having analysed the results of studies of the morphological composition of blood in 136 patients the author established that adaptation reactions (e. g. stress, training, activation) in their identification according to the hematologic values reflect the clinical course of the posttraumatic processes and may be used for the prognosis of the complications and for the control of the effectiveness of the treatment. PMID- 2571965 TI - Linkage of tyrosine kinase activity with transforming ability of the p185neu oncoprotein. AB - The neu oncogene encodes a 185 kd glycoprotein (p185neu) with intrinsic tyrosine kinase activity. Sequencing data has demonstrated that oncogenic p185neu differs from c-neu by a single point mutation within the transmembrane region of the glycoprotein. This mutation results in the substitution of a glutamic acid residue for a nonpolar valine residue at amino acid position 664 of the rat neu gene product. Recent studies have demonstrated that this mutation results in specific aggregation of the p185neu oncoprotein mimicking ligand induced dimerization events. The cellular consequences of the aggregated phenotype may include the enzymatic activation of p185neu. We demonstrate that the oncoprotein p185neu possesses higher intrinsic tyrosine kinase activity and that this increase in enzymatic activity is apparent within the plasma membrane, manifesting itself through the increased tyrosine phosphorylation of substrates. Furthermore, the neu oncoprotein itself is also phosphorylated on tyrosine to a higher extent than the proto-oncoprotein. These results strongly link enzymatic activation of p185neu to cellular transformation events. To test directly the effect of p185neu tyrosine kinase activity on cellular transformation we constructed mutant p185neu devoid of ATP binding ability. This mutant protein is expressed at high levels, but is unable to induce the transforming phenotype. The point mutation within the transmembrane region of p185neu mimics aspects of ligand induced activation events including increases in the specific tyrosine kinase activity of the molecule leading to cellular transformation. PMID- 2571966 TI - Expression of the c-erbB-2 gene product (p185) at different stages of neoplastic progression in the colon. AB - The c-erbB-2 gene has been found amplified in a number of human adenocarcinomas leading to elevated levels of expression of the p185 protein product. Increased expression of this putative growth factor receptor has been reported to occur by molecular mechanism other than gene amplification and for this reason we have studied the expression of the p185 protein in normal colon and in lesions representing different stages of neoplastic progression. We report amplification of the c-erbB-2 gene in 3 of 44 colon carcinomas and 1 of 5 preneoplastic polyps studied. Confirmation of expression of the p185 protein product was established in Western blot analysis and by immunocytochemical staining of tissue sections. An extended study, involving adenomatous polyps and carcinomatous material in immunostaining, revealed detectable presence of the p185 protein in 20% of carcinomas, consistent with immunoprecipitation data derived using established cell lines. In contrast, a high percentage of polyps showed strong staining with both p185 antibodies used, indicating elevated levels of expression of the c-erbB 2 protein associated with preneoplastic lesions. Staining of normal human colon revealed a restricted localization of this putative receptor to cells on the luminal colonic surface, with no expression in cells of the crypt. Histologically normal mucosa, adjacent to the tumor, showed a more extensive distribution involving the crypt suggestive of a disturbance in the normal expression of c erbB-2. These results indicate that elevated expression of the c-erbB-2 protein is associated with early stages of colonic neoplasia but do not establish it as a primary factor in these events. The occurrence of multiple copies of the c-erbB-2 in a percentage of colon lesions, however, suggests a possible role for this gene in some colon malignancies. PMID- 2571967 TI - [Relation of the chelating property of nitroxoline, the surface hydrophobicity and the inhibition of bacterial adherence]. AB - Nitroxoline or 5-nitro-8-hydroxyquinoline acts by a chelating effect with various metallic divalent cations. The chelating property of nitroxoline has been proposed as an hypothesis to explain the activity of this drug, at sub-MIC, on the inhibition of bacterial adherence. Nitroxoline (MIC/4) does not inhibit fimbriae synthesis but its antibacterial activity on E. coli 387 (MS/MS) was decreased by the addition of MgCl2 (50 mM) and CaCl2 (10 mM). The chelating effect of nitroxoline is mainly due to the presence of the nitrous radical in position 5. Nitroxoline would act at the outer membrane level of the bacterial cell-wall by a chelating effect preferentially with Mg++ than Ca++. Furthermore, nitroxoline (MIC/8) increases the bacterial surface hydrophobicity of E. coli 38 in contrast to EDTA (MIC/4). The both products inhibit the bacterial adherence to cells with the same manner. PMID- 2571968 TI - [Circadian variation of the nephrotoxicity induced in rats by bi-daily repeated administration of amikacin]. AB - We have studied the chrononephrotoxicity of amikacin during and after a prolonged treatment in rats. Animals received by intramuscular route a daily dose of 400 mg/kg (in two times) during 7 days at two different times (8.00 - 20.00 and 14.00 - 2.00). Large time-dependent variations in renal injury had been evidenced by several parameters and particularly by enzymuria. This injury is maximal when administration is made at 14.00 - 2.00. These data confirmed our results obtained in previous investigations with single daily injections and evidence that chrononephrotoxicity could permit an optimization of nephrotoxic drugs largely used in clinics. PMID- 2571969 TI - [Value of enzymuria during antibacterial therapy]. AB - Enzymuria is a well known parameter of evaluation of drugs nephrotoxicity, particularly of antibiotics. Alanine aminopeptidase (AAP), gamma glutamyltransferase (GGT) and N-acetyl-beta-D-glucosaminidase activities were measured in native urine. This study included 19 patients treated by an association of netilmicin-vancomycin. Enzymuria was measured on 24 hours urines at J0 then every two days during treatment. Enzymuria increased 24 or 48 hours after the beginning of the treatment. The Principal Components Analysis (PCA) of the results of enzymuria, seric urea and creatininemia shows the presence of two groups of responses. The first principal component exhibits two populations: the patients with pathological seric urea and pathological seric creatinine and the others. The PCA does not allow this discrimination using only the results of enzymuria; in contrast, with these results, the patients may be classified by the PCA on the basis of treatment duration. The enzymuria allows the clear identification of nephrotoxic drugs but does not allow the prediction of renal injury or of its aggravation. PMID- 2571970 TI - [Development of HIV 1 antigenemia during treatment with azidothymidine (AZT): follow-up of 90 patients for a year]. AB - VIH 1 antigenaemia has a significant value in the follow-up of patients treated with AZT. This study of 90 patients (55 ARC - 35 AIDS), each receiving AZT for more than a year, 200 mg every 4 hours, demonstrates the prognosis value of antigenaemia at Day 0, as well as its therapeutic indication value. However, at term and under this treatment, the significance of this virological data has to be reconsidered. Various kinetic patterns are described according to the clinical status and the CD4+ cells count. PMID- 2571971 TI - [Effect of induction of antibodies to serotonin on alcohol consumption and the status of the brain mediator system in rats with various degrees of alcohol motivation]. AB - Active immunization with serotonin-protein conjugate in rat experiments increased ethanol preference in animals with weak alcoholic motivation but reduced it in those with a high alcohol consumption. The level of antibodies to serotonin in blood was higher in alcohol-preferring rats than in water-preferring animals. The reduction of alcohol intake correlated with reduction of the level of 5-OH indoleacetic acid in the animals' hypothalamus. It is assumed that the contrary effect of immunization with serotonin-protein conjugate on the alcoholic behavior of animals with a different initial level of alcoholic motivation is linked with the difference in their neurochemical organization. PMID- 2571972 TI - Pharmacology of new hormonal therapies in the treatment of pediatric endocrine disorders. AB - Advances in genetic engineering will make possible treatment of many pediatric endocrine disorders with replacement therapy. Some of these conditions include short stature, precocious puberty, and diabetes mellitus. Although the availability of such hormonal replacement offers new treatment modalities, an understanding of their mechanism of action and pharmacologic characteristics is crucial to maximize their effectiveness while minimizing possible untoward effects. The clinician must evaluate potential risks and benefits as these substances come to market without definitive answers being available as to their long-term effects. PMID- 2571973 TI - Abnormal patterns of pulmonary neuroendocrine cells in victims of sudden infant death syndrome. AB - Ventilatory dysfunction has become the main focus of current research in sudden infant death syndrome (SIDS). This has been correlated with structural abnormalities in the carotid body and respiratory nuclei of the brainstem. In recent studies, the denervating effect of asphyxial brainstem dysfunction on the pulmonary neuroendocrine cells, which probably function as chemoreceptors, was demonstrated and prompted the following study. The pulmonary neuroendocrine system was evaluated in 25 victims of SIDS and 20 control infants, ranging in age from 3 weeks to 7 months and 1 to 12 months, respectively. The pulmonary neuroendocrine cells were stained by the Churukian-Schenk method and the neuroendocrine cell-positive airway values expressed as a percentage of the total number of airways. The range of positive airway values for victims of SIDS was 2% to 97% with a median of 73%. In contrast, the range for the control infants was 1% to 44% with a median of 25.5%. The SIDS victims' percentage was significantly greater than the control infants' percentage (P less than .0001). The number of pulmonary neuroendocrine cells in positive airway was also increased among SIDS victims compared with control infants. The altered pulmonary neuroendocrine cell pattern could be attributable to either brainstem dysfunction or chronic hypoxia. These explanations are not, however, mutually exclusive of one another; in fact, it is possible that both mechanisms may be operative. PMID- 2571975 TI - Sequence analysis of the D7S8 locus. PMID- 2571974 TI - Somatostatin activates an inwardly rectifying K+ channel in neonatal rat atrial cells. AB - Somatostatin, localized throughout the central and peripheral nervous systems, has been found in neurons of the vagal inhibitory pathway of the heart and has been shown to have negative inotopic effects in cardiac tissue. Using patch clamp techniques we show that somatostatin activates an inwardly rectifying K+ channel in rat atrial cells. Loss of somatostatin-induced K+ channel activity in excised inside-out patches is restored by the addition of GTP to the bath. Pertussis toxin pretreatment blocked GTP-dependent somatostatin activation of the inwardly rectifying K+ channel. This K+ channel has a conductance of 34 pS and a mean open time of approximately 1 ms. It is apparently the same K+ channel activated by muscarinic and adenosine receptors in atrial and cardiac pacemaker cells. Thus, atrial cells have at least three receptors which act via pertussis toxin sensitive G proteins to activate the same class of K+ channels. PMID- 2571976 TI - Isolation and mapping of a polymorphic DNA sequence (pCJ52.94T1) on chromosome 16 [D16S159]. PMID- 2571977 TI - A TaqI RFLP at the gas-2 locus. PMID- 2571978 TI - HincII RFLP at the human hexokinase I (HK1) locus on chromosome 10. PMID- 2571979 TI - BglII polymorphism in the human interleukin 6 (IL 6) gene. PMID- 2571980 TI - A PvuII RFLP in the human ADH2 gene. PMID- 2571981 TI - BamHI and EcoRI restriction fragment length polymorphisms at the glutathione S transferase 3 locus. PMID- 2571982 TI - RFLPs in human X-linked PGK1: a new probe for the PstI RFLP demonstrates strong linkage disequilibrium with the BgII RFLP. PMID- 2571983 TI - Isolation and mapping of a polymorphic DNA sequence (CJ52.10) on chromosome 16 [D16S153]. PMID- 2571985 TI - Isolation and mapping of a polymorphic DNA sequence (CJ52.197) on chromosome 16 [D16S156]. PMID- 2571984 TI - Isolation and mapping of a polymorphic DNA sequence (CJ52.1) on chromosome 16 [D16S152]. PMID- 2571986 TI - Isolation and mapping of a polymorphic DNA sequence (CJ52.105) on chromosome 16 [D16S154]. PMID- 2571987 TI - Isolation and mapping of a polymorphic DNA sequence (pCJ52.209M2) on chromosome 16 [D16S151]. PMID- 2571988 TI - Profile of in vitro binding affinities of neuroleptics at different rat brain receptors: cluster analysis comparison with pharmacological and clinical profiles. AB - A series of 21 neuroleptics with different chemical structures (phenothiazines, thioxanthenes, dibenzodiazepines, butyrophenones, benzamides, etc.) was examined for their in vitro interactions with 12 neurotransmitter binding sites in the rat brain (alpha- and beta-noradrenergic, dopaminergic, muscarinic, serotoninergic, histaminic, and opioid receptors, calcium channels, and serotonin uptake binding sites). The biochemical profile obtained from the binding data was compared with reported pharmacological and clinical profiles for this class of compounds by cluster analysis. Cluster analysis on binding data classified the compounds in three main subgroups: benzamides, compounds with an affinity mainly for DA2 and 5 HT2 receptors and inactive at muscarinic receptors, and compounds with a high affinity for alpha 1-adrenergic receptors and muscarinic receptors. The main subgroups resulting from cluster analysis of previously published pharmacological and clinical data for neuroleptics contain compounds common to the present study, with some correlations. The results extend previous observations that a complete binding profile corresponds to the pharmacological and clinical profile of this class of compounds. PMID- 2571989 TI - [Detection of hepatic metastasis of digestive cancers. Value of enzyme assays and ultrasonography]. AB - Alkaline phosphatase (AP), gamma-glutamyl transpeptidase (GGT), lactate dehydrogenase (LDH) and aspartate aminotransferase (AsT) assays, as well as ultrasonography are the easiest and least expensive examinations to perform in the diagnosis of hepatic metastases. The 273 patients included in this series had cancer of the digestive tract. The diagnosis of presence or absence of liver metastases was made at surgery and was positive in 38 patients (14 per cent). A receiver operating characteristic (ROC) curve was drawn after computing the sensitivity (Se) and specificity (Sp) of each laboratory determination while the threshold indicating that the value was normal was incremented. The examinations were then compared in terms of Se, Sp, positive predictive value and negative predictive value. The threshold was determined on the ROC curve where less false positive and more true-positive results were shown. According to predictive values, laboratory determinations could be classified in decreasing order of usefulness as: AP, LDH, GGT and AsT. Ultrasonography had a positive predictive value of 68 per cent a negative predictive value of 95 per cent, both figures being higher than those of any laboratory examination. These results suggest that ultrasonography has a higher diagnostic value than any of the enzyme assays in the detection of hepatic metastases. Moreover, ultrasonography provides morphological information which, in case of liver resection, may be useful to the surgeon. PMID- 2571990 TI - Synthesis of DNA containing the simian virus 40 origin of replication by the combined action of DNA polymerases alpha and delta. AB - Proliferating-cell nuclear antigen (PCNA) mediates the replication of simian virus 40 (SV40) DNA by reversing the effects of a protein that inhibits the elongation reaction. Two other protein fractions, activator I and activator II, were also shown to play important roles in this process. We report that activator II isolated from HeLa cell extracts is a PCNA-dependent DNA polymerase delta that is required for efficient replication of DNA containing the SV40 origin of replication. PCNA-dependent DNA polymerase delta on a DNA singly primed phi X174 single-stranded circular DNA template required PCNA, a complex of the elongation inhibitor and activator I, and the single-stranded DNA-binding protein essential for SV40 DNA replication. DNA polymerase delta, in contrast to DNA polymerase alpha, hardly used RNA-primed DNA templates. These results indicate that both DNA polymerase alpha and delta are involved in SV40 DNA replication in vitro and their activity depends on PCNA, the elongation inhibitor, and activator I. PMID- 2571991 TI - Induction of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase activity by fungal elicitor in cultures of Petroselinum crispum. AB - The effects of a cell wall fraction of the fungus Phytophthora megasperma on the enzymatic activities of 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase (EC 4.1.2.15) in extracts of cultured parsley cells (Petroselinum crispum) were examined. The specific activity of a plastidic form of DAHP synthase, designated DS-Mn by Ganson et al. [Ganson, R. J., d'Amato, T. A. & Jensen, R. A. (1986) Plant Physiol. 82, 203-210], was increased 2- to 3-fold in extracts of treated cells, with maximum induction occurring with 60 micrograms of fungal elicitor per ml after 6-8 hr. The cytosolic form of DAHP synthase, DS-Co, was unaffected by fungal elicitor. In the same experiments, phenylalanine ammonia lyase activity (EC 4.3.1.5) increased, while no effect on isoforms of chorismate mutase (EC 5.4.99.5) was observed. The subcellular localization of the two DAHP synthase isoforms in parsley was confirmed by differential centrifugation. Prior treatment of cultures with actinomycin D or cycloheximide prevented the increase in DS-Mn activity, indicating transcriptional regulation. PMID- 2571992 TI - Hemopoietic stem cells in murine embryonic yolk sac and peripheral blood. AB - Disaggregated embryonic yolk sac cells and circulating peripheral blood cells were obtained from normal murine day 9 embryos, prior to the formation of the fetal liver. These cells were microinjected transplacentally into days 11-15 W mutant anemic fetuses, when the fetal liver was the major hemopoietic organ. In a small proportion of the recipient animals examined after birth, long-term repopulation by the embryonic donor hemopoietic cells was observed. The donor hemopoietic stem cells proliferated and differentiated in the hosts as evidenced by the presence of donor hemoglobins in the growing recipient host animals. Some mothers of the pups were also repopulated by the donor stem cells. These results provide direct evidence that, during early murine embryogenesis, there are functional hemopoietic stem cells which are capable of colonizing the adult hemopoietic organs and probably the fetal liver and spleen to initiate hemopoiesis in these tissues. PMID- 2571994 TI - Prostaglandin E1 in systemic sclerosis. PMID- 2571995 TI - Effects of linoleic and alpha linolenic acids intake on blood pressure in man. AB - The effect of dietary linoleic and alpha-linolenic acid was evaluated in 18 healthy females on LFDs. Polyunsaturated fatty acids had little effect on blood pressure, n-6 fatty acids decreased and n-3 fatty acids increased blood pressure. PMID- 2571993 TI - Genetic analysis of the imperfect association of H-2 haplotype with lupus-like autoimmune disease. AB - Unlike parental New Zealand Black (NZB) or New Zealand White (NZW) mice, (NZB x NZW)F1 mice exhibit a lupus-like disease characterized by high serum levels of IgG anti-nuclear antibodies and a fatal immune complex glomerulonephritis. Previous results from studying [(NZB x NZW)F1 x NZB] backcross mice indicated that the NZW major histocompatibility complex (MHC) or gene(s) closely linked to this locus provides the major dominant NZW genetic contribution to the F1 disease. A surprising feature of the results was the 12% frequency of discordance between the autoimmune phenotype and the presence of the NZW H-2z haplotype. In the current study, we attempted to precisely define the position of the NZW gene(s) required for lupus-like renal disease by mapping genes in individual backcross mice that are both centromeric and telomeric to the MHC and then correlating genotypes for each locus with disease. The data indicate that an adjacent NZW locus does not provide a more accurate correlation with the autoimmune phenotype compared with MHC genes themselves. Thus, the imperfect association of MHC haplotype with disease in this murine model is not explained by genetic recombination with linked genes. These data may provide insight into the mechanisms by which MHC antigens increase the probability of developing autoimmune disease and may help explain the difficulty of defining MHC relationships in human systemic lupus erythematosus. PMID- 2571996 TI - Effect of H1-antihistamines and calmodulin antagonists on the ionophore A23187 induced eicosanoid release from human leukocytes. PMID- 2571997 TI - Typing of MHC haplotypes in OS chicken by means of RFLP analysis. AB - For the analysis of the genetic background of autoimmune thyroiditis we used the Obese strain (OS) chicken model which develops a SAT. Practically all animals from this strain show severe lymphoid infiltration of the thyroid gland and circulating autoantibodies against thyroglobulin (Tg-AAb) within a few weeks after hatching. Of the 3 MHC haplotypes (B5, B13, B15) present in the OS, B13 was mostly associated with severe thyroid infiltration. Haplotypes B5 and B15 were associated both with severe, as well as with mild infiltration. To clarify these controversial results published by different groups and to further assess the role of the MHC in the development of SAT, we selected by appropriate breeding sublines with high and low levels of Tg-AAb. With the help of serological methods and GvH assays we were not able to find additional differences in the MHC antigens of that line. Therefore, for further characterization of these haplotypes, RFLP analysis was applied in the present study. Southern blots were done with restriction enzyme digests of erythrocyte DNA hybridized with a chicken cDNA probe (code-p234) for MHC class II antigens. The Southern blots with BamH-I digests showed at least 5 bands, four of which were polymorphic. Four RFLP patterns emerged, two of which were observed within chickens with the B15 haplotype. The confirmation of this RFLP heterogeneity within serologically identical haplotypes requires additional analysis. PMID- 2571998 TI - Analysis of five B complex haplotypes with class I (B-F), class II (B-L) and class IV (B-G) probes. PMID- 2571999 TI - The ocular effects of eicosanoids and other autacoids: historic background and the need for a broader perspective. AB - Great progress has been made in describing the synthesis of most known eicosanoids by ocular tissues. This progress has included identification of the spectrum of eicosanoids synthesized by different ocular tissues of different species, as well as comparisons of the relative proportion of individual eicosanoids produced by ocular tissues of different species. In contrast to this wealth of information, the lack of antagonists that are specific to individual eicosanoids has greatly hindered efforts to determine the specific ocular functions of each member of this family of autacoids. Furthermore, the absence of high-affinity ligands that are specific for individual eicosanoid receptors has greatly limited our ability to classify these receptors and to assign specific eicosanoid effects to specific cell types. In the early years of ocular PG research, the lack of recognition of the existence of parallel pathways of arachidonic acid metabolism and the lack of appreciation of the effects of some other autacoids--most notably the recently discovered neuropeptides--led to the premature conclusion that PGs have mediatory roles in a number of ocular pathophysiological responses. More recent studies have demonstrated that some ocular effects, such as chemotaxis, that were first attributed to classical PGs of the E and F type are actually mediated by eicosanoids synthesized by the lipoxygenase pathway, whereas other ocular responses, at least in some species, are mediated by various neuropeptides. These neuropeptides can be released by the same stimuli that cause the release of arachidonic acid. Furthermore, interpretation of the ocular effects of eicosanoids as well as neuropeptides has been complicated by large species differences in the effects of these autacoids on intraocular tissues. Among mammalian species, the rabbit eye and the primate eye may represent opposite extremes with respect to the extent of their responses to irritation and trauma, as well as with respect to their responses to some autacoids, especially eicosanoids and neuropeptides. In spite of these differences, the rabbit remains the animal most widely used in eye research. Consequently, many of the findings reviewed in this volume are also based on studies done only on rabbits. However, in view of the foregoing, we must resist the temptation to extrapolate to primates results obtained only from studies of rabbit eyes.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2572000 TI - A physiological approach to glaucoma management: the use of local hormones and the pharmacokinetics of prostaglandin esters. AB - It appears that a pragmatic approach, based entirely on the ability of a drug or procedure to reduce IOP, has thus far been used to develop modalities for the management of glaucoma. Barring serendipity, it is unlikely that this approach will produce effective new methods for the management of this ocular disorder, which presents an ever-increasing problem in an increasingly longevous and industrialized world population. Although a better understanding of the biology of aqueous humor dynamics and its pathogenesis, combined with new genetic engineering techniques, may eventually lead to the total elimination of glaucoma, that ultimate solution is unlikely to be achieved within the foreseeable future. Thus, a physiologic approach to glaucoma management must be developed, one that takes into consideration all of our recently acquired knowledge of aqueous humor dynamics and all other relevant physiologic principles. A review of currently available information suggests that from a physiologic point of view the best approach to glaucoma management, at least for the next few decades, will be based on the use of receptor-mediated, naturally occurring ocular hypotensive agents, or at least agents that closely resemble such naturally occurring autacoids. Based on the temporal and spatial limitations of the various classes of receptor mediated autacoids and on differences in the specificity and the nature of the responses they elicit at different sites, we conclude that a physiologic approach to glaucoma management should focus on the use of topically applied local hormones or their analogs. This approach appears to be feasible in light of demonstrations that several members of one family of local hormones, the eicosanoids that are produced within ocular tissues under physiological conditions are effective ocular hypotensive agents. It has been shown that one member of this family, PGF2 alpha, in its esterified form, is an extremely potent ocular hypotensive agent in its esterified prodrug form in both normotensive and glaucomatous human eyes, yielding significant IOP reduction when topically applied in doses less than one-hundredth that of currently available glaucoma medications. Although no intraocular side effects have been noted in human eyes after topical application of PGF2 alpha-1-isopropylester in submaximal ocular hypotensive doses, such side effects as conjunctival hyperemia and foreign body sensation present a problem and a challenge.4 PMID- 2572001 TI - Studies on D1 and D2 dopamine receptor involvement in conditioned taste aversions. AB - This series of studies investigated the ability of compounds selective for either the D1 or D2 dopamine receptor to induce a conditioned taste aversion (CTA) in thirsty rats. Neither the D1 antagonist SCH23390 (0.12-0.60 mg/kg) nor the D2 antagonist haloperidol (0.125-0.375 mg/kg) were able to induce CTAs to a saccharin solution. In contrast, the D1 agonist SKF38393 produced a dose dependent taste aversion which was stereoselective to the (R-) enantiomer. The aversion to (R,S)-SKF38393 was not blocked by pretreatment with either SCH23390 or haloperidol, suggesting that the aversion is not mediated through stimulation of either dopamine receptor subtype. The D2 dopamine receptor agonist quinpirole was also found to produce a dose-dependent CTA. This aversion was blocked by injections of haloperidol and was attenuated following injections of domperidone, suggesting involvement of peripheral dopamine receptors in the aversion. Pretreatment with SCH23390 failed to affect the quinpirole-induced CTA, providing additional evidence that the D1 and D2 dopamine receptor subtypes can function independently of one another in the control of behavior. Finally, it does not appear that the area postrema is importantly involved in these taste aversions since lesions of this brain region did not affect the CTAs induced by either SKF38393 or quinpirole. PMID- 2572002 TI - Effects of the putative anxiolytic SM-3997 on central monoaminergic systems. AB - The effects of SM-3997 on central monoaminergic systems were evaluated by ex vivo measurement of monoamines and their metabolite levels in rat brain after intraperitoneal treatment of drugs and by in vitro measurement of monoamine uptake into rat brain slices. The effects of SM-3997 were also compared with those of other new nonbenzodiazepine anxiolytic compounds. SM-3997, buspirone, gepirone and ipsapirone showed no effects on serotonin uptake and dopamine uptake, and a weak inhibition of norepinephrine uptake at the concentration of 100 microM. SM-3997 decreased the serotonin metabolite (5-hydroxyindole-3-acetic acid) level without changing the serotonin level in hippocampus and increased dopamine metabolite (3,4-dihydroxyphenylacetic acid, homovanillic acid) level with no effect on the dopamine level in striatum. SM-3997 also produced an increase in the norepinephrine metabolite (3-methoxy-4-hydroxyphenylglycol) level with a decrease in the norepinephrine levels in hippocampus. Similar effects on serotonin metabolites and norepinephrine metabolites were observed in several other regions. Although the serotonergic effect of SM-3997 was similar to that of buspirone, gepirone and ipsapirone, the dopaminergic effect of SM-3997 was much weaker than that of buspirone. PMID- 2572003 TI - Acute and subchronic effects of methylenedioxymethamphetamine [(+/-)MDMA] on locomotion and serotonin syndrome behavior in the rat. AB - Specific behaviors comprising the serotonin syndrome (low body posture, forepaw treading, headweaving) and the autonomic signs of piloerection and salivation were determined and analyzed with locomotor activity in response to MDMA at three doses (2.5, 5.0, and 7.5 mg/kg). All behaviors were dose-responsive. Serotonin syndrome behaviors increased in both intensity and duration of response with increasing doses. In contrast, locomotion varied only in intensity. Subchronic injections, in the same group of animals, permitted an analysis of acute vs. subchronic effects on these same behaviors. Both the serotonin syndrome and locomotor behaviors were augmented on subsequent testing, indicating that, (+/ )MDMA, like amphetamine, is capable of producing behavioral sensitization. PMID- 2572004 TI - Circling evoked by intranigral SKF 38393: a GABA-mediated D-1 response? AB - Intranigral injection of microgram doses of the dopamine D-1 receptor agonist SKF 38393, in rats acutely anaesthetised with halothane, did not overtly alter the animals' behaviour. The adenylate cyclase activator forskolin, the GABA uptake inhibitor nipecotic acid and the GABA potentiator pentobarbital, were similarly ineffective when administered singly to one substantia nigra pars reticulata. However, all three treatments interacted synergistically with coinjected SKF 38393 to promote active circling. It is suggested this SKF 38393-induced behavioural response is mediated by GABA released from D-1 receptor-bearing striatonigral neurones. PMID- 2572005 TI - Effects of nonsedating histamine H1-antagonists on EEG activity and behavior in the cat. AB - The central effects of the newly-developed antihistamines (H1-receptor antagonists) loratadine, astemizole, mequitazine and terfenadine were evaluated by studying brain electrical activity (EEG), sleep-waking patterns and behavior in the cat. The different stages of the sleep-waking cycle, i.e., wakefulness (W), spindle sleep (SS), slow wave sleep (SWS) and REM sleep (REM) were evaluated. The power spectrum analysis of the EEG was obtained by a computerized technique. For comparison, the sedating agent diphenhydramine was examined. Given at 3 mg/kg orally, a dose slightly above that effective therapeutically, diphenhydramine markedly affected behavior and all sleep stages. In particular, it depressed REM and increased SS (drowsiness). The EEG showed occasional spikes typical of subconvulsive states. Loratadine did not modify either sleep patterns or behavior over the 3-30 mg/kg dose range orally, which is far above that used clinically. The EEG, evaluated either visually or by spectral power analysis, was unaffected. Astemizole at 10 and 30 mg/kg PO reduced REM, markedly altered behavior at 30 mg/kg, but did not modify EEG activity. Mequitazine, at low doses (1-10 mg/kg PO), enhanced SS and decreased SWS and REM. Like diphenhydramine, mequitazine induced EEG changes typical of subconvulsive states and affected EEG power over the frequency range of 0.1-15.0 Hz. Terfenadine did not change sleep patterns and slightly affected behavior only at the high dose of 30 mg/kg orally; EEG activity was not influenced. These data show that: a) diphenhydramine and mequitazine appear to produce CNS effects by altering basic processes within the brain; b) astemizole and terfenadine seem to cross the blood-brain barrier at high doses only; c) loratadine has the lowest liability to produce central side effects. Of the sleep features examined, REM appeared to be the most sensitive stage to blockade of central H1-receptor pathways. PMID- 2572006 TI - Possible role of alpha-2 and alpha-1 adrenoceptors in the experimentally-induced depression of the central nervous system. AB - The alpha-2 adrenoceptor agonists clonidine and xylazine were employed in chicks and rats to induce a loss of the righting reflex, a sign for depression of the central nervous system. These effects of clonidine and xylazine were antagonized by yohimbine, idazoxan and CH-38083 (7,8-(methylenedioxi)-14-alpha hydroxyalloberbane HCl), compounds having alpha-2 adrenoceptor antagonist properties. Prazosin, an antagonist for alpha-1 adrenoceptors, enforced the alpha 2 adrenoceptor agonist-induced depression in both species. 6-Hydroxydopamine treatment, which reduced the norepinephrine concentrations in the rat cerebral cortex by 76%, increased the duration of the loss of righting reflex induced by xylazine indicating that central postsynaptic alpha-2 adrenoceptors might also be involved in this behavioral alteration. The electrically-stimulated tritium release was also determined from the isolated rat cerebral cortex slices which had been preloaded with 3H-norepinephrine. Clonidine and xylazine inhibited the stimulation-induced tritium release and this inhibition was counteracted by yohimbine, idazoxan or CH-38083, but not by prazosin. We have concluded from the present data that stimulation of alpha-2 adrenoceptors with pre- and postsynaptic locations or inhibition of alpha-1 adrenoceptors in the central nervous system may shift the depression/vigilance balance to the direction of depression which might be accompanied by a decreased activity of cortical noradrenergic neural transmission. PMID- 2572007 TI - Opposite effects of SK&F 38393, a dopamine D-1 agonist, and SCH 23390, a dopamine D-1 antagonist, in tests of salt preference/aversion in the rehydrating rat. AB - Male rats were adapted to a 22-hr water-deprivation schedule, and to a 15-min choice test, in which water was available in one drinking tube, and water, 0.064%, 0.16%, 0.4%, 1.0%, or 2.5% NaCl solution, respectively, was available in a second. A typical saline preference-aversion function was obtained. The selective dopamine D-1 agonist, SK&F 38393 (3.0 mg/kg, IP), significantly depressed choice of hypertonic saline solutions (1.0% and 2.5% NaCl solutions), without affecting preference for hypotonic saline solutions. In contrast, the selective dopamine D-1 antagonist, SCH 23390 (0.1 mg/kg, SC), significantly increased the preference measure in the case of hypertonic solutions. These data indicate a role for D-1 receptors in dopaminergic mediation of the descending limb of the saline preference-aversion function. PMID- 2572008 TI - The effects of amfonelic acid alone and in combination with naloxone on brain stimulation reward. AB - Thresholds for rewarding brain stimulation delivered to the medial forebrain bundle-lateral hypothalamus were determined by means of a rate-free psychophysical method. Amfonelic acid (AFA), an indirect dopamine agonist, alone caused a significant dose-dependent lowering of the rewarding threshold. Although naloxone treatment by itself did not significantly alter the reward threshold, it blocked AFA's threshold lowering effect in every animal at one or more of the dose combinations of the two drugs tested. Naloxone was found to be more effective in blocking the threshold lowering actions of a higher (1 mg/kg) dose as opposed to a lower (0.25 mg/kg) dose of AFA. Since a lowering of threshold for rewarding intracranial stimulation is a model for drug-induced euphoria, the findings presented here indicate that AFA may have abuse potential. Furthermore, these results suggest that endogenous opioid systems may begin to modulate the effects of AFA on the reward system only when a certain level of activation of dopaminergic systems is reached. PMID- 2572010 TI - Estimation of the incidence of HIV infection. AB - The aim of the method of 'back projection' is to provide estimates of the number of new infections with the human immunodeficiency virus (HIV) as a function of time, by using the numbers of diagnoses of the acquired immune deficiency syndrome (AIDS) together with information on the distribution of the incubation period between infection and diagnosis. Here, the method is investigated with particular reference to cases of HIV infection and AIDS in the United Kingdom. PMID- 2572009 TI - Milacemide increases 5-hydroxytryptamine and dopamine levels in rat brain- possible mechanisms of milacemide antimyoclonic property in the p,p'-DDT-induced myoclonus. AB - Milacemide, a glycine prodrug that is able to enter the brain readily, has been shown to have an antimyoclonic property in the p,p'-DDT-induced myoclonus syndrome. Milacemide increased regional 5-HT and dopamine and decreased 5-HIAA, DOPAC and HVA levels in naive rats. In p,p'-DDT-treated rats, 5-HT levels were unchanged at the time the rats experienced spontaneous myoclonus in all brain regions except in the striatum, where it increased. 5-HIAA levels increased but did not reach significant levels except in the striatum. Dopamine, DOPAC, HVA and norepinephrine were unchanged. When rats were treated concurrently with both p,p' DDT and milacemide, regional 5-HT levels were increased and NE levels in the brainstem and cerebellum decreased. Depletion of brain serotonin by pretreatment with PCPA or with 5,7-DHT, or blocking 5-HT receptors with different 5-HT antagonists, failed to eliminate the antimyoclonic property of milacemide. This antimyoclonic effect of milacemide may be mediated through other mechanisms besides its ability to increase brain 5-HT levels. Possible mechanisms to be considered are its antiepileptic property, and its ability to increase brain glycine levels. Milacemide may have potential for therapeutic trials in patients with myoclonus. PMID- 2572011 TI - Predictions of the AIDS epidemic in the U.K.: the use of the back projection method. AB - Back projection methods are used to predict the yearly number of new AIDS diagnoses and the number of new HIV infections, to the end of 1992. The AIDS, but not the HIV, predictions are insensitive to the choice of incubation period distribution. A wide range of predictions is consistent with the AIDS diagnoses in years up to 1987, but limited ancillary information on the relative rates of new HIV infection in 1984 to 1987 can be used to narrow this range. The range of prediction based on AIDS reports to the end of 1988 is lower and narrower than the range based on reports to the end of 1987. The number of new AIDS cases in 1992 appears likely to fall in the range 1000-3000. PMID- 2572012 TI - A process of events with notification delay and the forecasting of AIDS. AB - Analysis and prediction of a point process are studied when there is delay in notifying the occurrence of events. A primarily parametric approach is taken to studying the rate of the underlying process and to predicting future properties of the system. Data on AIDS are used in illustration. PMID- 2572013 TI - The overall distribution of survival times for U.K. AIDS patients. AB - Data on the survival times of 997 U.K. AIDS patients are analysed with the aim of deriving a simple form for the overall survival distribution. The exponential and Weibull distributions are modified to accommodate specific features of the data, in particular, the recording of survival times to the nearest month and the occurrence of a significant proportion of cases recorded as having zero time on study. The final model has a probability 0.08 of underlying survival time being zero and, given non-zero survival time, takes the form of an exponential distribution with mean of 14.95 months. The results are in close agreement with those of a study of New York City patients as well as the empirical data. PMID- 2572014 TI - Evidence for recent changes in sexual behaviour in homosexual men in England and Wales. AB - Over 80% of cases of Acquired Immune Deficiency Syndrome (AIDS) in England and Wales have occurred in homosexual men. Changes in sexual behaviour in this group may have a substantial influence on the incidence of Human Immunodeficiency Virus (HIV) infection and will therefore be crucial in determining future cases of AIDS. This paper critically weighs the indirect and direct evidence for changes in behaviour in homosexual men since the advent of the AIDS epidemic. The paper reports on falling incidence of gonorrhoea, hepatitis B and syphilis in homosexual men, the changes being most marked from 1985 onwards. Data on temporal trends in HIV prevalence and incidence in homosexual men are reviewed. These suggest that the maximum incidence of HIV infection occurred in 1982-84 and may have fallen since then. Evidence for a concomitant change in sexual behaviour is reported from several sources. This points towards a recent change in sexual behaviour characterized by reduction in the numbers of partners and adoption of safer sexual practices. In some places change may have occurred as early as 1983. A change became apparent generally in 1985 and this appears to have been sustained in 1986-87. Nevertheless, a substantial proportion of homosexual men studied continue to practice high risk sexual practices, such as anal intercourse, including relationships with casual partners. PMID- 2572015 TI - Estimating the size of the HIV epidemic by using mortality data. AB - Evidence that more people are dying as a result of HIV infection than is reflected by the number of deaths among reported cases meeting the WHO definition of AIDS is derived from mortality data. Ninety-five causes of death likely to be associated with HIV infection were selected. Standardized mortality ratios due to these causes increased for single men aged 15-54 years from 100 in 1984 to 118 in 1987. The age, sex, marital status, temporal and geographic distribution of these excess deaths suggest that they are HIV-associated. It is estimated that 58% of excess deaths due to HIV-related causes were among cases reported to the CDSC AIDS Surveillance Programme in 1987. Some of these deaths may have been among HIV positive people who did not meet the WHO definition at the time of death. There is a need for surveillance to be extended to include HIV-positive people who die before meeting the WHO definition if the full extent of the HIV epidemic is to be identified. PMID- 2572016 TI - Rate of growth of AIDS epidemic in Europe: a comparative analysis. AB - Estimates of the rate of increase of the AIDS epidemic for each of 18 European countries are obtained by fitting a Poisson process with exponential rate of growth to data. A linear regression model of these estimates on the proportion of cases that are intravenous drug users, homosexuals/bisexuals and heterosexuals, was estimated and suggested that the rates of growth of the epidemics amongst these groups are different and in increasing order. Empirical Bayes estimates of the rates are obtained for each country. PMID- 2572017 TI - Seropositivity for HIV in U.K. haemophiliacs. AIDS Group of the United Kingdom Haemophilia Centre Directors. AB - The results of a third seroprevalence survey of antibody to HIV in U.K. haemophiliacs, done in 1987, are presented. Out of 3028 men with haemophilia A living in the U.K. during the period 1980-1987 who had been tested for HIV antibody, 39% were positive, and out of 517 men tested with haemophilia B, 5% were positive. For both haemophilia types the proportion of seropositive men is greater among patients with a severe coagulation defect. No new seroconversions are known to have taken place after November 1986. PMID- 2572018 TI - Prediction for small subgroups. AB - Prediction limits are calculated for the number of events likely to occur in a specified time period in an exponentially growing epidemic. The basis for the prediction is the total number of events observed in the past. PMID- 2572019 TI - Epidemiological and statistical aspects of the AIDS epidemic. PMID- 2572020 TI - Epidemiological and statistical aspects of the AIDS epidemic: introduction. AB - In 1988, a government working party studied estimates of incidence and prevalence of numbers of acquired immunodeficiency syndrome (AIDS) cases. They investigated a series of epidemiological, statistical and mathematical problems associated with predicting trends in incidences of AIDS. This paper introduces a series of papers that give a fuller and more technical exposition of the appendixes of that working party report. The papers provide a brief background to the current state of knowledge on the epidemiology of the infection and the disease; a deterministic model for human immunodeficiency virus (HIV) transmission in the male homosexual community in England and Wales is introduced. Back-projection methods are studied in two papers, following the distribution of the incubation period of the disease. The concept of minimum size of the epidemic is introduced. Mathematical functions to describe the spread of HIV infection are refined by using past trends in the incidence of AIDS to estimate values for some parameters. Survival times for AIDS patients from the point of diagnosis are considered and evidence for changes in male homosexual sexual behaviour is studied; lag-time from the point of diagnosis to the report of the case is also examined. There is a comparative analysis of the AIDS epidemic in various European countries. The incubation period of HIV in patients with haemophilia A and B infections and the problems associated with making predictions for different at-risk groups or small subgroups based on geographical area are discussed. Reasons for fluctuation between the number of reported cases from month to month are provided. PMID- 2572021 TI - The transmission dynamics of the human immunodeficiency virus type 1 in the male homosexual community in the United Kingdom: the influence of changes in sexual behaviour. AB - This paper examines the transmission dynamics of human immune deficiency virus type 1 (HIV-1) in the male homosexual population in the U.K. via numerical studies employing a mathematical model representing the principal epidemiological process. The model is based on an assumption of proportionate mixing between different sexual-activity classes (defined by the rate of sexual partner change per unit of time) and incorporates heterogeneity in sexual activity, distributed infection and incubation periods and the recruitment of susceptibles to the sexually active population. The sensitivity of model predictions to various assumptions and parameter assignments is examined. Numerical studies of model behaviour focus on the influence of changes in the magnitudes of the transmission parameters, associated with three periods of infectiousness during the incubation period of acquired immune deficiency syndrome (AIDS), on the magnitude and duration of the epidemic and on the level of the endemic equilibrium state. Predicted temporal trends in the incidence of AIDS are shown to be particularly sensitive to changes in the intensities and durations of the stages of infectiousness. Most of the paper addresses the influence of changes in sexual behaviour on the magnitude and duration of the epidemic. Numerical simulations show that the manner in which behavioural changes occur and who is influenced by such changes (i.e. infecteds or susceptibles, the sexually active population or new recruits to this population) have a major impact on the future timecourse of the epidemic. The greatest reduction in the incidence of AIDS over the coming decades is induced by changes in the rate of sexual-partner change among the sexually active population, particularly those currently infected. The time periods at which changes in behaviour occur, in relation to the starting point of the epidemic (assumed to be 1979), are also of particular significance to the future pattern of the incidence of disease and infection. Changes in behaviour early on in the timecourse of the epidemic have a much greater impact than equivalent changes at latter time points. On the basis of limited data on the pattern of change in sexual behaviour among the male homosexual community in the U.K., numerical studies of model behaviour tentatively suggest that the epidemic is at, or near to, a period of peak incidence of the disease AIDS. Analyses suggest that, following the peak in incidence, there will be a period of slow decline over many decades provided recent changes in behaviour are maintained in the coming years.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2572022 TI - Population projections for AIDS using an actuarial model. AB - This paper gives details of a model for forecasting AIDS, developed for actuarial purposes, but used also for population projections. The model is only appropriate for homosexual transmission, but it is age-specific, and it allows variation in the transition intensities by age, duration in certain states and calendar year. The differential equations controlling transitions between states are defined, the method of numerical solution is outlined, and the parameters used in five different Bases of projection are given in detail. Numerical results for the population of England and Wales are shown. PMID- 2572023 TI - [Long-term benzodiazepine medication]. AB - Benzodiazepine (BD) misuse and dependence in 80 patients, 1974-1983 undergoing withdrawal treatment, were investigated by means of case histories and catamnestic inquiries. RESULTS: Three fourth of the patients were women. Many of the patients suffered from disturbed matrimonial relations, and 36 lived unmarried, divorced, or widowed. Besides, among the men frequently vocational difficulties were found, which increased in the course of BD-taking. In particular after removal of Lorazepam or Bromazepam in 58 cases withdrawal symptoms appeared, among them seven times delirium and six times epileptic seizures (grand mal). Catamnestic inquiries were practicable in 51 patients. 20 of them had required no further treatment, while 18 took again BD. But only 3 still proved to be as well psychical as physical dependent, in contrast to 14 before withdrawal treatment. Among the 15 only psychical dependent patients 8 merely showed a low dose dependence. Hence BD-dependence proves to be remediable, and subsequently shows only little tendency to relapse. PMID- 2572024 TI - How do the neuropathological changes of schizophrenia relate to pre-existing neurotransmitter and aetiological hypotheses? PMID- 2572025 TI - Cerebrospinal fluid somatostatin in delirium. AB - Cerebrospinal fluid somatostatin-like immunoreactivity (CSF SLI) was determined for 67 elderly patients who met the DSM-III criteria for delirium and for 19 age matched controls. As a group, and also when subdivided according to the type of delirium, severity of cognitive decline or the type of central nervous system disease, the delirious patients showed significant reductions of SLI compared with the controls, together with a declining trend associated with increasing cognitive dysfunction. These findings are in accordance with previous observations that reduced CSF SLI is associated with diseases in which cognitive function is disturbed and they extend this finding to delirium. PMID- 2572026 TI - Use and abuse of khat (Catha edulis): a review of the distribution, pharmacology, side effects and a description of psychosis attributed to khat chewing. AB - There have been relatively few reported cases of psychosis due to khat usage despite its heavy consumption in certain East African and Arabian countries. Four cases have been reported in the UK. We report here on three further cases of psychotic reactions to this substance in Somalian males, and emphasize the need to be aware of khat as a potential substance of abuse, with both medical and psychiatric complications. The features of khat psychoses are described and the relationship to amphetamine and ephedrine psychoses is discussed. The forensic aspects of two of the cases which involved homicide and combined homicide and suicide are highlighted, as is the possible role of social dislocation from the culture of origin. PMID- 2572027 TI - SCH 23390 blocks drug-conditioned place-preference and place-aversion: anhedonia (lack of reward) or apathy (lack of motivation) after dopamine-receptor blockade? AB - The influence of the D1 antagonist SCH 23390 on the motivational properties of rewarding (morphine, nicotine and diazepam) and aversive (naloxone, phencyclidine and picrotoxin) drugs was studied in the rat in a two-compartment place conditioning paradigm, which included a pre-conditioning test for spontaneous place-preference. The specific D1 dopamine-receptor antagonist SCH 23390 (0.05 mg/kg SC), paired with both compartments or, separately, with the preferred or with the non-preferred compartment, failed to affect the spontaneous unconditioned preference of the animal. Pairing of morphine (1.0 mg/kg SC), nicotine (0.6 mg/kg SC) or diazepam (1.0 mg/kg IP) with the less preferred compartment induced significant preference for that compartment. Pairing of SCH 23390 (0.05 mg/kg SC) with both compartments completely blocked the place preference induced by morphine, nicotine and diazepam. Naloxone (0.8 mg/kg SC), phencyclidine (2.5 mg/kg SC) or picrotoxin (2.0 mg/kg IP) paired with the preferred compartment elicited place-aversion. Pairing of SCH 23390 (0.05 mg/kg SC) with both compartments abolished also the place-aversion induced by naloxone, phencyclidine and picrotoxin. The results indicate that blockade of dopamine transmission blocks the motivational properties of rewarding as well as aversive stimuli. It is suggested that neuroleptics rather than simply blocking the rewarding impact of positive reinforcers (anhedonia, lack of pleasure) exert a more general influence on conditioned behaviour by blocking the affective impact of negative as well as positive reinforcers (apathy, lack of motivation). PMID- 2572029 TI - Acute laryngeal dystonic reactions to neuroleptics. AB - Two cases of acute laryngeal dystonia (laryngospasm), a rarely reported extrapyramidal reaction to neuroleptics, occurred in a public psychiatric hospital. A review of the literature revealed only seven well-documented case reports. This article discusses the clinical significance of this rare, alarming, and probably underreported phenomenon. Factors related to recognition, prediction, and management are also discussed. The review strongly advocates immediate intravenous administration of anticholinergic drugs to relieve dystonia. PMID- 2572030 TI - Differentiating neuroleptic-related heatstroke from neuroleptic malignant syndrome. PMID- 2572028 TI - Blockade of dopamine receptors reverses the behavioral effects of endogenous enkephalins in the Nucleus caudatus but not in the Nucleus accumbens: differential involvement of delta and mu opioid receptors. AB - We have previously (Dauge et al. 1988) demonstrated that injection of the mu agonist [D-Ala2, MePhe4, Gly-ol5]-enkephalin (DAGO) or the delta agonist [D-Thr2, Leu5]-enkephalyl-Thr6 (DTLET) into the rat Nucleus accumbens (N.Acc.), or Nucleus caudatus (N.Caud.) induced a hypoactivity followed by hyperactivity 150 min later in the case of the mu agonist and a hyperactivity in the case of the delta agonist. Moreover, naloxone reversible delta-type responses were obtained by local infusion of kelatorphan, ([(R)-3(N-hydroxylcarboxamido-2-benzylpropanoyl)-L alanine]), a complete inhibitor of enkephalin catabolism, suggesting a tonic control of the behavioral activity of rat by the endogenous opioid peptides. In this work, the putative involvement of the dopaminergic system in these behavioral responses was investigated by using the DA antagonist thioproperazine. In the N.Acc., the behavioral effects of kelatorphan or of mu or delta agonists were not altered by thioproperazine-induced blockade of dopamine receptors. In contrast, the hyperactivity produced by DTLET or by kelatorphan in the N.Caud. was reversed by thioproperazine while the time-dependent biphasic effect resulting from DAGO injection remained unaffected by the DA antagonist. This blocking effect of thioproperazine is in agreement with the previously described delta-selective enhancement of the release of newly synthesized DA in the striatum but not in the N.Acc. PMID- 2572031 TI - [Accidents in young children in Flanders. Place of dental accidents in epidemiologic studies]. AB - Accidents requiring professional intervention were registered during 1 year in a population of nearly 6,500 Flemish children younger than 3 years. First, some professional records placed the children in their social context. Each month the parents were interviewed about the circumstances of the accidents, the need for professional help, the injuries, etc. Nearly 25% of the children had an accident during this study, needing professional help. The most frequent causes were falling (55.4%), cutting (11.4%), crushing (8.9%) and burning (7.6%). The most dangerous places were the kitchen and the living room (50%); the most frequent injuries were the open wound (42.2%), contusions (20.1%) and burns (7.8%). Only 5.6% needed the help of a family doctor. Only 2.8% of the accidents had consequences for the mouth and teeth, mostly following a fall on a level surface; 75% of these needed a dentist. The records were compared with other studies in Western Europe. Only the structure of the family (one or two parents) had an influence on the records. PMID- 2572032 TI - [Reliable laparoscopic pattern in polyarteritis nodosa (PAN). Its histologic corroboration and a new endoscopic contribution using photogastroscopy]. PMID- 2572033 TI - Exploring the bacterial surface and floating on the Dead Sea. PMID- 2572034 TI - Broxaterol: a new beta2-adrenoceptor agonist for the treatment of reversible airways disease. Proceedings of the satellite symposium. Budapest, September 8, 1988. PMID- 2572035 TI - Broxaterol: pharmacological profile of a unique structural feature. AB - A series of 1-(3-substituted-5-isoxazolyl)-2-alkylaminoethanol derivatives was synthesized in order to evaluate the effectiveness of the isoxazole ring in replacing the catechol moiety of beta-adrenergic compounds. Among the investigated compounds, 1-(3-bromo-5-isoxazolyl)-2-tert-butyl aminoethanol hydrochloride, named broxaterol hydrochloride (laboratory code Z 1170), resulted to be the most potent and selective beta 2-agonist of the series. In vitro studies confirmed the beta 2-agonist profile of broxaterol, which showed a marked bronchodilating activity in different experimental models. Interestingly, besides its direct effect in relaxing bronchial smooth muscle, broxaterol was, also, very effective in inhibiting asthmogenic mediator release both in vitro and in vivo. The preclinical studies demonstrated that the similar potency observed in vitro for both broxaterol and salbutamol resulted in a higher effectiveness of broxaterol with respect to salbutamol, when the compounds were given by oral route. These findings account for a probably greater bioavailability of broxaterol after oral administration. PMID- 2572036 TI - Broxaterol, a new beta 2-adrenoceptor agonist compared to salbutamol in asthmatics, oral and inhalation treatment. AB - Two double-blind, double-dummy, randomized, crossover studies were performed in 8 asthmatic patients to evaluate beta 2-adrenoceptor selectivity and potency of broxaterol compared to salbutamol. By oral route, a study with cumulatively increasing doses (total dose broxaterol 1.675 mg and salbutamol 26 mg) showed that as a bronchodilator broxaterol was 12-16 times more potent than salbutamol. The same potency difference was seen for an increase of heart rate and decrease of diastolic blood pressure, indicating that the clinical selectivity for beta 2 adrenoceptors is equal for the two drugs. One study with cumulatively increasing doses of broxaterol and salbutamol (total dose 1.5 mg for both) showed that with the inhaled route, broxaterol was somewhat less potent than salbutamol with respect to bronchodilation. Side effects, increase of heart rate and tremor were considerably less pronounced by the inhaled route as compared to the same bronchodilatory effect given by the oral route. PMID- 2572037 TI - Role of broxaterol in bronchial hyperresponsiveness. AB - The protective effect of broxaterol against bronchial hyperresponsiveness has been assessed in several double-blind randomized controlled studies in stable asthmatic patients. In three cross-over studies, broxaterol (400 micrograms by metered dose inhaler) has been shown to significantly reduce methacholine responsiveness 30, 60 and 120 min after dosing. The protective effect was similar to that obtained with terbutaline (500 micrograms), but there was a trend toward a higher activity of fenoterol (400 micrograms), even if counterbalanced by a more frequent occurrence of side effects. Broxaterol 0.5 mg by oral route provided a significant greater protection against the exercise-induced bronchospasm than 0.02 mg of clenbuterol and a placebo. Also in children, broxaterol 0.25 mg as nebulizer solution significantly prevented exercise-induced bronchospasm versus placebo, and this effect was seen to be associated with an improvement in maximal working capacity. Prevention of bronchospasm following 3 or 4 consecutive challenges with inhaled distilled water mist was demonstrated after oral and aerosol administration of broxaterol. This effect was not significantly different from that observed with salbutamol, if administered by aerosol. Finally, in a cross-over placebo-controlled study, broxaterol (400 micrograms by aerosol) significantly inhibited allergen-induced immediate bronchoconstriction. These data suggest that broxaterol represents a new valid alternative to the most effective beta 2-stimulants now commonly used. PMID- 2572038 TI - Broxaterol: therapeutic trials and safety profile. AB - The efficacy and the safety profile of broxaterol have been assessed in multicenter open studies and in some double-blind controlled clinical trials. Broxaterol (0.6-1.2 mg/day by metered dose inhaler or 0.5-1.5 mg/day orally, from 2 weeks to 1 year) produced a significant clinical improvement, an increase in FEV1 and a decrease in supplemental anti-asthmatic drugs used in patients with reversible airflow obstruction and in asthmatic children. The increases in FEV1 versus baseline were significantly maintained after the end of the treatment. Prompt disappearance of the asthmatic attack with significant improvement in lung function was observed in children. In two long-term controlled trials the respiratory effects of broxaterol nebulizer solution were significantly greater than placebo. Moreover, broxaterol by metered dose inhaler was more effective than salbutamol after 3 months follow-up, showing absence of tachyphylaxis. In long-term clinical evaluation, broxaterol has been shown to be well tolerated, with an incidence of adverse reactions equal to or less than that reported in the literature for other beta 2-agonists. The side effects most frequently associated with broxaterol were tremor, nervousness and palpitations. They usually appeared to be slight, transient and dose-related, requiring withdrawal from clinical trials in only 12 patients out of 274 (4.4%). Clinically relevant changes in heart rate and blood pressure were never reported. Broxaterol was found not to modify the ECG or haematological, hepatic and renal laboratory tests. No metabolic abnormalities or hypokalemia were caused by long-term treatment with broxaterol. PMID- 2572039 TI - Today's treatment of airway obstruction ... and tomorrow's? AB - Airflow limitation results from loss of elastic recoil (as in emphysema), or narrowing of large and small airways from smooth muscle contraction, mucosal swelling and/or oedema, mucous plugging (as in asthma), and loss of small airways (as in COPD). Bronchodilator regimes in asthma include inhaled beta 2-agonists, but these do not reduce bronchial hyper-reactivity but act quickly and synergise with oral slow-release theophyllines (serum level 10-20 micrograms/ml), without potentiating tremor which occurs with oral beta 2-agonists. Airway inflammation as the mechanism of asthma, although fashionable, remains unproven, and clearly requires to be specified for asthma. Inhaled steroids slowly improve FEV1 in asthma and in 10-20% of COPD, and reduce hyper-reactivity. Nedocromil has yet to reveal similar potency. Clinical trial of effective anti-PAF drugs or anti leukotriene agents are awaited, but understanding the specific asthmatic inflammation is still needed for rational therapy. In COPD combining inhaled beta 2-agonists with ipratropium--both given by a reservoir device--can be effective, along with oral slow-release theophylline and possibly inhaled steroids. New inhalation devices (i.e. modified dry powder inhalers) will be needed as the freon propellents in MDI may soon cease manufacture due to potential environmental hazards. PMID- 2572040 TI - Remarks on broxaterol. PMID- 2572041 TI - The pulsed dye laser in the treatment of ureteral calculi. PMID- 2572042 TI - [The case from general practice (143). Patient: Mr. F.-S. H. P., a 51-year-old baker]. PMID- 2572043 TI - [The family physician and drug side-effects. Observations from general practice- conclusions for general practice]. AB - Several side effects of drugs observed in an office of a general practitioner are described. The cause of these side effects, their prevention and their treatment are discussed. PMID- 2572044 TI - [Type I multiple endocrine neoplasia--Wermer syndrome]. AB - Multiple endocrine neoplastic diseases are genetically determined conditions with particular organ patterns for endocrine tumors. In Type I or Wermer's syndrome the endocrine pancreas, anterior pituitary and parathyroids are involved, insulinoma being the most frequent pancreatic tumor. To facilitate diagnosis, a prolonged oral glucose tolerance test, a fasting test and determination of the glucose-insulin ratio are recommended. Localisation is sought by computer tomography and angiography. A gastrinoma is excluded on the basis of normal gastrin levels in serum and by means of the secretin-provocation-test. Pituitary tumors can be classified more closely with prolactin levels and releasing-hormone tests (LH-RH and TRH). Prolactinoma is the most frequent pituitary tumor and amenable to bromocryptin treatment. If Wermer's syndrome is suspected, primary hyperparathyroidism has to be excluded on the basis of calcium and parathormone levels. Chief cell hyperplasia or multiple adenomas are frequent. Surgical resection is necessary. PMID- 2572045 TI - [Takayasu's disease and cerebromeningeal hemorrhage]. AB - A 53-old-woman with both Takayasu disease and intracerebral hemorrhage is reported. Non-specific aorto-arteritis was acquired, while aneurysm on arteria cerebri media is probably of congenital origin. The association of these two syndromes is, judging by this facts, accidental. Nevertheless, it should be taken into consideration than panarteritis had a significant role in formation of aneurysm and favored the appearance of hemorrhage. PMID- 2572046 TI - Treatment of pancreatic fistulas with somatostatin and total parenteral nutrition. AB - Nineteen pancreatic fistulas were treated with somatostatin (ST) and total parenteral nutrition (TPN). Five of the fistulas developed in an uninflamed pancreas, whereas 14 fistulas developed secondary to a necrotizing or chronic pancreatitis. Fistular output varied between 20 and 800 ml/day (median, 160 ml) during TPN before ST treatment; amylase concentration was 10,500-800,000 UI/l. Twelve of 16 (75%) fistulas were contaminated with bacteria. Thirteen of 19 (68%) fistulas closed after a median treatment of 7 (range, 2-14) days. Seven of eight fistulas with open drainage to the bowel healed, whereas only one of six with obstructed drainage closed. All of the uninfected fistulas and half of the infected fistulas closed. The findings suggest that somatostatin treatment speeds up the closure of pancreatic fistulas with open drainage to the bowel but is not beneficial when the intestinal drainage of the fistular region to the bowel is obstructed. PMID- 2572048 TI - [Lithotripsy with a laser, a look toward the future?]. AB - In the last few years the treatment of urinary calculi has been dramatically transformed by new endoscopic methods and extracorporeal shock wave lithotripsy. Laser lithotripsy is in keeping with this evolution. After a short description of the principle of laser lithotripsy, a prototype pulsed dye laser lithotriptor, developed by the Centre d'applications laser, Ecole polytechnique federale de Lausanne, is presented. This device is compared with other lasers proposed for the same application. PMID- 2572047 TI - Effects of mesalazine substitution on salicylazosulfapyridine-induced seminal abnormalities in men with ulcerative colitis. AB - Some semen characteristics of eight male patients with clinically inactive ulcerative colitis were investigated. Semen analysis was carried out twice during salicylazosulfapyridine (SASP) treatment and repeated twice after at least 3 months' treatment with mesalazine. The motility variables all showed significant improvement during mesalazine treatment: the graded motility (p less than 0.05), motility in percentage (p less than 0.01), and the penetration in egg white (p less than 0.05). The semen plasma was analyzed for mesalazine and the metabolite Ac-mesalazine during both regimens. There was no difference in the semen plasma concentration of mesalazine during the two regimens, whereas Ac-mesalazine was significantly higher during mesalazine treatment than during SASP treatment, indicating that other SASP metabolites, most likely sulfapyridine, are the agent causing the abnormal sperm characteristics. We suggest that pure mesalazine preparations are a safer alternative in young men with chronic inflammatory bowel disease. PMID- 2572050 TI - [The retained testicle: our experience]. AB - In our experience the undescended testis has an occurrence rate of 3-4% on the total of males born. The rate decreases to 0.3% when the occurrence is observed on males aged 1 year. The first step of treatment is hormonal followed by a surgical second step in case of failure. The Author stresses the need of the treatment as early as possible to prevent the damage of both exocrine and endocrine testicular functionality. PMID- 2572049 TI - Neurofibromatosis and inflammatory bowel disease. PMID- 2572051 TI - [Cryptorchism and fertility. Our experience]. AB - The relationship between cryptorchidism and fertility has been suggested in a lot of ways but hasn't been clearly demonstrated. In our Institute from 1981 until 1987, 352 orchidopexies had been performed. Out of these, 31 patients were studied with respect to the current age, the age at the surgery, the absence of varicocele and the possible iatrogenic damages. The rate of dispermia among the subjects studied, was 43% whereas 57% had normospermia. 3 out of 4 patients operated for bilateral undescended testis were azoospermic. The volume of unaffected testis is directly related to the health of the seminal status. PMID- 2572053 TI - [Cryptorchism: long-term results in subjects evaluated in a conjugal infertility service (monolateral cryptorchism fertility)]. AB - Infertile male subjects because of cryptorchidism have an overall rate of 0.5-1%. Undescended testis even if treated by surgery and/or hormonal therapy have a bad seminal map. However in Author's experience, the hormonal treatment give a significant improvement in terms of fertility. Biopsies carried out on undescended testis demonstrated an early cellular damage of the testis. PMID- 2572052 TI - [Cryptorchism yesterday and today. Update study programmed in interdisciplinary services]. AB - It is more than 25 years that several Authors stress the need of an early treatment of undescended testis both mono or bilateral. The pathogenesis of undescended testis is still unknown, but some suggestions about the hormonal role has been formulated. From these considerations "interregional groups to study the cryptorchidism" has been formed, in order to use the same parameters of evaluation: auxologic studies, cytogenetic analysis, hormonal tests, dynamic hypophyseal functionality test, medical treatment, surgical treatment, biopsies, postoperative hormonal treatment, seminal evaluation after 20 years of life. From all these studies a protocol has been planned, including a treatment by HCG associated with urophollitropine as pure FSH stimulating effect and an eventual surgical treatment before the 5th years of life. PMID- 2572054 TI - [Testicular biopsy in cryptorchism: prognostic parameters]. AB - 470 testicular biopsies performed in prepuberal cryptorchides has been reviewed in order to assess the relationship, if any, between the morphological parameters and the prognosis of future fertility. The main parameter is the presence and the number of germinal cells, which are progressively decreasing in the following groups of patients: descended testis in monocryptorchid patients, monolateral undescended testis, bilateral undescended testis. The results observed are similar to those reported by various Authors. The age of the patients biopsied was irrelevant with respect to the results of the predictability of fertility. Other parameters than germinal cells are of no practical interest. PMID- 2572056 TI - [Fertility in the operated on monolateral cryptorchism patient]. AB - Fertility of operated monolateral Cryptorchides has been evaluated in a long term follow up comparing the patients who underwent orchidopexy with patients who underwent orchiectomy. From our experience, as well as from various Author's reports, patients who underwent orchiectomy demonstrated a two fold of fertility compared to the group who underwent orchidopexy. PMID- 2572055 TI - [Indications for the hormonal therapy of cryptorchism]. AB - The treatment of cryptorchidism is nowadays based on one of the following drugs: chorionic gonadotropins, FSH, LHRH analogues. The chorionic gonadotropin treatment is the first choice treatment given in quite similar way by various Authors. It is furthermore the less expensive. Its activity is mainly pharmacological. The chorionic gonadotropin can be administered both as monotherapy as well as associated. The associated treatments are with FSH and with LHRH analogues. The latter way of treatment requires a skilled experience because of the precise diagnosis needed to administer the LHRH analogues. PMID- 2572057 TI - [Intranasal gonadorelin in the treatment of cryptorchism]. AB - In 94 outpatients medium aged 4.16 years the LHRH analogues Gonadorelin had been administered for 4 weeks. 74 had monolateral cryptorchidism whereas 18 had bilateral cryptorchidism. The results indicate that Gonadorelin has a role in treating cryptorchidism. The Authors suggest, on its experience, that the Gonadorelin should be the first choice of treatment of cryptorchidism. PMID- 2572058 TI - [Gastric secretion blockaders in peptic ulcer]. PMID- 2572059 TI - Lipoproteins during antihypertensive therapy. Study supported by the Swiss National Science Foundation. PMID- 2572060 TI - Risk factors as they relate to drug therapy in hypertension. PMID- 2572061 TI - Molecular characterisation of a low-frequency mutation in exon 8 of the human low density lipoprotein receptor gene. AB - The prevalence of familial hypercholesterolaemia (FH), an autosomal dominant disease characterised by raised low-density lipoprotein (LDL) cholesterol levels, is at least five times higher in the white Afrikaner population than in most other population groups in the world. A founder gene effect has been suggested to explain this abnormally high frequency. Detection of a polymorphic Stu I site in the 5' region of the LDL receptor gene and association of both restriction fragment length polymorphism alleles with FH in Afrikaners, indicated the existence of at least two founder members for the disease in this population. DNA from a hetero-allelic FH homozygote from this South African group has been analysed through genomic cloning and sequencing. The DNA polymorphic site is caused by a single guanine to adenine transition within exon 8 of the LDL receptor gene and can be used in the determination of haplotype-associated defects. PMID- 2572062 TI - [Vascular permeability and microcirculation in patients with hemorrhagic fever associated with renal syndrome]. AB - Vascular permeability and microcirculation (MC) were studied in 25 patients with hemorrhagic fever associated with the renal syndrome (HFRS). A considerable increase in vascular permeability for liquid and protein was recorded during oligoanuria and polyuria. These parameters slowly returned to normal with recovery. Investigation of MC identified the presence of changes mainly occurring in the intra- and extravascular parts of MC: turbidity of the angioscopic background, hemorrhages, spread sludge, atonia of the venules, winding of all groups of the vessels, aneurysms of the venules, and so forth. The increased vascular permeability and MC impairment are important components of the pathogenesis of HFRS which should be taken into consideration during institution of the pathogenetic treatment. PMID- 2572063 TI - Elevated expression of proto-oncogenes accompany enhanced induction of heat-shock genes after exposure of rat embryos in utero to ionizing irradiation. AB - We have recently found that the effects of exposing rat embryos in utero to teratogens capable of producing cardiac anomalies were expressed later as enhanced induction of heat-shock proteins (hsp70 family) when embryonic hearts were cultured in vitro. However, it remained to be determined whether heat-shock proteins are induced in vivo after exposure to teratogens. The heat-shock response in some mammalian systems is known to be accompanied by elevated expression of proto-oncogenes. Using gene-specific DNA probes, we examined the levels of the expression (transcription) of heat-shock protein genes and two nuclear proto-oncogenes, c-fos and c-myc, in the embryos removed from irradiated pregnant mother rats 4 or 5 days after the irradiation. We found that the levels of expression in vivo of the hsp70 and c-myc genes in the irradiated embryos increased by approximately twofold as compared with those in the control. The expression in vivo of the c-fos gene was not detected in either the irradiated or non-irradiated embryos. After 0.5-hr incubation in vitro of the embryos, however, the expression of the c-fos gene in the irradiated embryos was highly enhanced whereas the control showed no changes. Although the exact functions of these gene products still remain obscure, the enhanced expression of hsp70 gene(s) and the nuclear proto-oncogenes observed in the present study may reflect repair of intracellular damages and/or regeneration of tissue by compensatory cell proliferation, processes that may disturb the normal program of organogenesis. PMID- 2572065 TI - Hydrogen peroxide mutagenicity towards Salmonella typhimurium. AB - In bioassays conducted under controlled, comparable conditions, weak direct mutagenicity responses were observed for hydrogen peroxide in the standard (Ames test) agar plate incorporation bioassay with Salmonella typhimurium strains TA97, TA98, TA102, and TA1537, in a 20 min preincubation test with strains TA97, TA98, TA100, TA102, TA1537, and TA1538, and in a liquid incubation modification using strain TA1537. These results conclusively demonstrate that hydrogen peroxide is a weak mutagen, especially in strains that are sensitive to oxidative damage under suitable bioassay conditions. PMID- 2572064 TI - Cell cycle alterations and cell death in cyclophosphamide teratogenesis. AB - Litters of pregnant mice treated with cyclophosphamide (CP) exhibit malformations of the limbs ranging from oligodactyly to amelia. Previous studies have indicated that cell death occurs in limb buds shortly after maternal exposure. We have investigated the relationship of cell death, cell cycle perturbation, and embryo/fetal toxicity in the mouse using vital staining and flow cytometry (FCM). CP (20, 30, and 40 mg/kg) was investigated via intraperitoneal administration to Swiss-Webster mice on day 10 of gestation. At 4, 8, or 28 hours later, embryos were removed. Cell death was identified with Nile blue sulphate (NBS). Two embryos per litter were stained with NBS, and the remaining embryos were frozen at -70 degrees C prior to FCM analysis. After thawing, the forelimb buds were removed for the isolation of nuclei. Tissues were dissociated through a wire mesh followed by cytolysis with 0.1% nonidet P-40 in PBS with 0.5 mg/ml RNase. Nuclei were stained with the fluorescent nucleic acid probe propidium iodide and analyzed (10,000 nuclei per sample) for propidium iodide fluorescence by FCM. NBS revealed a dose-related increase in cell death by 8 hours after dosing. CP induced cell death was greatest in areas of rapid cell proliferation (DNA synthesis). FCM analysis revealed retardation of progression through the S-phase of the cell cycle by 4 hours post-exposure at all doses. This retardation occurred earlier in S-phase with increasing dose and persisted through 8 hours. At 28 hours, cell cycle histograms were normal in the low-dose embryos, but remained perturbed in the intermediate- and high-dose embryos. On day 17 of gestation, the last group of dams was killed. A high incidence of fetal malformations, including limb defects, occurred at the 20 mg/kg dose, and fetal mortality was observed at 30 and 40 mg/kg. The pattern and magnitude of cell death correlated with cell cycle perturbation and fetal toxicity at term, suggesting a relationship between cell cycle perturbation, cell death, and malformations produced by CP. PMID- 2572066 TI - Induction of sister chromatid exchange in multiple murine tissues in vivo by various methylating agents. AB - In order to study the reliability of in vivo sister chromatid exchange (SCE) assays for predicting carcinogenicity, several known animal carcinogens were tested in a multicellular in vivo SCE assay and an in vivo/in vitro murine lymphocyte assay. The methylating agents 1,2-dimethylhydrazine.2 HCl (DMH), dimethylnitrosamine (DMN), methylnitrosourea (MNU), methyl methane-sulphonate (MMS), and methylazoxymethanol acetate (MAM) were tested for SCE induction in several murine tissues in vivo, including bone marrow, alveolar macrophages, regenerating and intact liver, and kidney from B6D2F1 mice. In all cell types, clear dose-responses were observed following exposure of mice to subcytotoxic fractions of the LD50 dose of DMH, MNU, or MMS. DMN (0.03-0.27 mmol/kg) produced small, although not dose-related, increases in SCE in all cell types. At the doses tested (0.06 and 0.08 mmol/kg), MAM did not induce elevated SCE in the various cell types. Following a series of multiple i.p. injections of low, non toxic doses of DMH (0.15 mmol/kg, once a week, for 10 weeks), significant differences were observed in intact vs. regenerating liver and in single vs. multiple injections in regenerating liver. Following exposure of B6D2F1 mice to a single i.p. injection of 0.25 mmol/kg DMN, DMH, or MMS or 0.19 mmol/kg MNU, SCE responses were evaluated in Concanavalin A (Con A)- and LPS-stimulated blood and spleen lymphocytes. Considerable cytotoxicity was observed in blood lymphocytes. In Con A- and LPS-stimulated spleen lymphocytes, DMH-, and DMN- and MMS-induced SCE frequencies were approximately 1.5-2 x baseline levels and MNU-induced SCE were approximately three- to fourfold higher than baseline values in cultures initiated at 1 and 24 h postexposure. At 48 and 72 h after an i.p. injection of 0.131 mmol/kg MNU, SCE responses in lymphocytes were approximately 2 x baseline levels. At 24 h following one, two, or four injections (one/week) of 0.075 mmol/kg MNU dose-related increases in SCE were observed in spleen lymphocytes. These studies illustrate that carefully designed in vivo SCE assays may have the capacity to predict the tumorigenic potential of chemical agents. PMID- 2572068 TI - Embryotoxic and teratogenic studies of phosphamidon in Swiss albino mice. AB - Phosphamidon, an organophosphate pesticide, is little known for its possible effects on mammalian conceptus. The present investigation was carried out to evaluate its embryotoxic and teratogenic effects on Swiss albino mice. Female mice of similar age and weight were divided into four experimental groups. The animals of groups I and II received 15 and 35 ppm phosphamidon ad libitum, respectively, during the entire gestational period, that is, from days 1 to 18. Groups III and IV were treated before mating with 35 ppm phosphamidon for 30 and 60 days, respectively, and the treatment was continued during pregnancy up to the 18th day. The autopsies were performed on the 18th day of gestation, and routine teratological observations were made. The lower dose did not produce significant effects. The higher dose reduced the number of implants, litter size, and foetal weight and increased the resorption of embryos when administered for 30 days prior to mating. Exposure to the pesticide for 60 days prior to mating and during gestation, however, did not produce significant effects. It appears that the mice probably developed some resistance to the pesticide, when exposed for such a long duration. PMID- 2572067 TI - Micronucleus formation by benzene, cyclophosphamide, benzo(a)pyrene, and benzidine in male, female, pregnant female, and fetal mice. AB - Male, female, pregnant female, and fetal ICR mice were compared for their acute sensitivity to four single doses of model carcinogens, as measured by micronucleus formation in polychromatic erythrocytes 24 h after treatment in adult bone marrow and fetal liver at days 17-19 of gestation. Cyclophosphamide caused a dose-responsive increase in micronuclei in all groups, without a consistent difference based on gender or pregnancy. At doses of 50 and 75 mg/kg given orally to the pregnant female, the fetuses were three to six times as sensitive as was the mother. Benzo(a)pyrene showed a similarly increased sensitivity of the fetus relative to the other groups, although it is a much weaker clastogen. Benzidine did not cause an increase in micronuclei in any group, although it was thought that the fetal liver might have been sensitive enough to detect it, relative to adult bone marrow. Benzene caused much less response in females than in males and almost no response in pregnant females and their fetuses, even though pregnant females metabolized at least half as much of the total dose as did the males (as measured by the presence of urinary metabolites of benzene). No single metabolite of benzene in the urine was consistently correlated with micronucleus formation in the bone marrow. Several factors must be interacting in different ways for different chemicals to influence their clastogenicity. PMID- 2572069 TI - How often are polymorphic restriction sites due to a single mutation? AB - An approximate expression is obtained for the probability that a restriction site, which is polymorphic in a random sample, is a site at which two or more mutations have occurred in the descent to the sample from the most recent common ancestor of the sample. The analysis is based on the assumption that the population from which the sample is obtained is at equilibrium under a selectively neutral Wright-Fisher model. Monte Carlo simulations show that the approximation is quite accurate. For commonly observed levels of genetic variation in humans and in natural populations of Drosophila, it is found that multiple mutations would occur at 5 to 10 percent of polymorphic restriction sites assuming that six-cutter enzymes are used on samples of size 50 to 100. Simulations are also used to investigate the bias and mean square error of four estimators of 4Nu, where N is the population size and u is the neutral mutation rate per nucleotide site. Two of the estimators are biased by approximately 20 percent when levels of variation are similar to those which have been observed in natural populations of Drosophila. PMID- 2572070 TI - [Drug therapy of pain]. AB - Pain represents the most frequent complaint for which patients ask for medical help. Successful treatment of acute and chronic pain syndromes largely depends on the knowledge and the therapy of the underlying disease process and on the rational use of analgesics, such as the nonnarcotic analgesic-antipyretics, the anti-inflammatory agents and the opioid analgesics. In addition, so-called 'adjuvant' analgesics of different chemical structures can be used in special clinical conditions to increase the efficacy of the common analgesics. For optimal therapy, the selection and dosing of a specific agent has to be tailored to the individual needs of the patient. For this purpose it is important to follow some general principles of pain management and to consider the relevant pharmacokinetic and pharmacodynamic properties of the various analgesics. This overview summarizes the most important pharmacological properties of the widely used analgetic drugs, with special emphasis on their risk-benefit ratio in various clinical situations. PMID- 2572072 TI - The use of DNA amplification for genetic counselling related diagnosis in haemophilia B. AB - A single base pair variation in the coding sequence of coagulation factor IX produces a protein polymorphism detectable with monoclonal antibodies and a restriction fragment length polymorphism (RFLP). This allows carrier and prenatal diagnoses in 48% of Caucasian families segregating for haemophilia B. However, this RFLP cannot be detected by standard Southern blotting, while the antibody assay may give equivocal results in some females and can only allow prenatal diagnoses on second trimester fetal blood samples. We show that, using the polymerase chain reaction, the polymorphic DNA segment can be amplified and directly tested for the presence of the alternative sequences by a non radioactive procedure that has the advantage of speed (1-2 days), partial automation and applicability to first trimester diagnoses. We also show that the method gives results on a single drop of dried blood. PMID- 2572071 TI - [Allergic bronchial asthma and allergic rhinitis: clinical aspects and therapy]. AB - Bronchial asthma and allergic rhinitis are the most common chronic diseases in childhood. On the bases of bronchial hyperreactivity many factors can trigger asthma. In children at preschool and school age allergy plays the most important role. The typical asthmatic attack is rarely missed, but frequently recurrent, irritant coughs at night or after physical exercise are misinterpreted. In the present article clinical picture, diagnosis and today's treatment of allergic asthma are discussed. The most important therapy consists of inhalation with efficacious drugs at the time of an acute attack and is continued as a prophylaxis in cases of perennial asthma. Allergic rhinitis does fortunately not take the threatening course of asthma, but its symptoms can reduce the well-being and the quality of life of a child. The diagnosis of allergic rhinitis is easy and rarely missed. The current therapy - like asthma-therapy - consists of avoidance of any known provoking factors in addition to topical application of drugs interacting with mast-cells and corticosteroids and to oral non-sedative histamines. PMID- 2572073 TI - Isolation and structural characterization of a potent inhibitor of coagulation factor Xa from the leech Haementeria ghilianii. AB - The present work reports the discovery and characterization of an anticoagulant protein in the salivary gland of the giant bloodsucking leech, H. ghilianii, which is a specific and potent inhibitor of coagulation factor Xa. The inhibitor, purified to homogeneity, displayed subnanomolar inhibition of bovine factor Xa and had a molecular weight of approximately 15,000 as deduced by denaturing SDS PAGE. The amino acid sequence of the first 43 residues of the H. ghilianii derived inhibitor displayed a striking homology to antistasin, the recently described subnanomolar inhibitor of factor Xa isolated from the Mexican leech, H. officinalis. Antisera prepared to antistasin cross-reacted with the H. ghilianii protein in Western Blot analysis. These data indicate that the giant Amazonian leech, H. ghilianii, and the smaller Mexican leech, H. officinalis, have similar proteins which disrupt the normal hemostatic clotting mechanisms in their mammalian host's blood. PMID- 2572074 TI - [Cryptorchism in dogs]. AB - The outstanding features of cryptorchidism in dogs, including the diagnosis, clinical symptoms and breeding policy, are discussed. PMID- 2572075 TI - Clinical and laboratory findings implicating palytoxin as cause of ciguatera poisoning due to Decapterus macrosoma (mackerel). AB - A near fatal case of ciguatera-related intoxication following consumption of smoked Decapterus macrosoma is documented. In addition to some of the hallmark symptoms of ciguatera poisoning, the patient exhibited acute respiratory distress and severe muscle spasms. Laboratory results showed large elevations in a number of blood enzymes, indicative of muscle damage. The responsible agent was extracted from corresponding fish samples and identified as palytoxin. PMID- 2572076 TI - Purification and characterization of the 180-kDa membrane guanylate cyclase containing atrial natriuretic factor receptor from rat adrenal gland and its regulation by protein kinase C. AB - The original concept that cyclic GMP is one of the mediators of the hormone dependent process of steroidogenesis has been strengthened by the characterization of a 180-kDa protein from rat adrenocortical carcinoma and rat and mouse testes. This protein appears to have an unusual characteristic of containing both the atrial natriuretic factor (ANF)-binding and guanylate cyclase activities, and appears to be intimately involved in the ANF-dependent steroidogenic signal transduction. In rat adrenal glands we now demonstrate: 1) the direct presence of a 180-kDa ANF-binding protein in GTP-affinity purified membrane fraction as evidenced by affinity cross-linking technique and by the Western blot analysis of the partially purified enzyme; 2) that the enzyme is biochemically and immunologically different from the soluble guanylate cyclase as there is no antigenic cross-reactivity of 180-kDa guanylate cyclase antibody with soluble guanylate cyclase; 3) in contrast to the soluble guanylate cyclase, the particulate enzyme is not stimulated by nitrite-generating compounds and hemin; and 4) protein kinase C inhibits both the basal and ANF-dependent guanylate cyclase activity and phosphorylates the 180-kDa guanylate cyclase. These results reveal the presence of a 180-kDa protein in rat adrenal glands and support the contention that: (a) this protein contains both the guanylate cyclase and ANF receptor; (b) the 180-kDa enzyme is coupled with the ANF-dependent cyclic GMP production; (c) the 180-kDa enzyme is biochemically distinct from the nonspecific soluble guanylate cyclase; and (d) there is a protein kinase C-dependent negative regulatory loop for the operation of ANF-dependent cyclic GMP signal pathway which acts via the phosphorylation of 180-kDa guanylate cyclase. PMID- 2572077 TI - Biological activity of damavaricin derivatives. PMID- 2572078 TI - Epidemiologic background of blood donors with antibody to human T-cell lymphotropic virus. Transfusion Safety Study Group. AB - We interviewed 51 blood donors in four major US metropolitan areas subsequently found to have had antibodies to human T-cell lymphotropic virus (anti-HTLV) in late 1984-early 1985. Sixteen donors (31%) reported that they or a sexual contact had a history of blood transfusion. Twelve donors (24%) reported that they or a sexual contact used intravenous drugs. Ten donors (20%) were blacks born in the southeastern US. Four of the male donors (15%) reported homosexual contact. The most common characteristic was an association with Japan or the Caribbean basin (61%). These results show a broader variation of epidemiologic backgrounds than anticipated. PMID- 2572079 TI - HIV-2 in Spain. PMID- 2572080 TI - Molecular chaperones: proteins essential for the biogenesis of some macromolecular structures. AB - Many polypeptides can self-assemble into functional structures while others assemble only in the presence of additional proteins (molecular chaperones) which are not components of the final structure. We discuss here the effect that the recognition of the essential roles played by these proteins in assembly processes may have on the principle of spontaneous self-assembly. PMID- 2572081 TI - PQQ, the elusive coenzyme. AB - The recently discovered redox coenzyme, PQQ (methoxatin), is widely distributed. Quantitation of protein-bound PQQ has been difficult, but unique redox cycling reactions, which reflect its striking biological properties, reveal trace amounts. PQQ is a potential target for drugs. PMID- 2572082 TI - B lymphocyte response as an indicator of acute renal transplant rejection. I. Immunoglobulin-secreting cells in peripheral blood. AB - Monitoring of immunoglobulin-secreting cells in peripheral blood was performed in 88 renal transplant recipients using a reverse hemolytic plaque-forming cell assay. Comparison with other in vitro tests for rejection (plasma neopterin, CD4/CD8 ratio) demonstrated that the number of immunoglobulin-secreting cells in peripheral blood provides a highly sensitive rejection marker. Evidence of rejection was obtained 1.7 +/- 0.4 (mean +/- SEM) days before a rise in creatinine, with a significant PFC rise in 95% (73/77) of rejection episodes. The PFC response was not influenced by HLA matching, number of preoperative blood transfusions, acute tubular necrosis, or uremia. A significant PFC rise in the absence of an ongoing rejection episode occurred in the presence of bacterial or viral infections, in case of posttransplant surgical complications, and regularly during the early posttransplant period (days 4-9). However, even early posttransplant the PFC peak was significantly higher in patients with an ongoing rejection episode than in patients without rejection (P less than 0.001). PMID- 2572083 TI - Requirements for the induction of allospecific CD8+ suppressor T cells in the rat primary mixed lymphocyte response. CD4+, CD45R+ T cells, or supernatant factor. AB - We examined the requirements for the induction of the MLR-generated allospecific CD8+ suppressor T cells in the rat. Depleting the responder population of CD4+ T cells before initiating the primary MLR abrogates the generation of day-5 CD8+ T suppressor effectors. Readdition of at least 10% CD4+ T cells to the CD4+ depleted primary MLR reconstitutes suppressor cell generation. Using the anti CD45R monoclonal antibody OX22, we also show that the T suppressor inducer cells are CD4+ CD45R+. Using a dual chamber Transwell culture system, which allows cells to be co-incubated without direct cell-to-cell contact, we show that a soluble factor/s, produced during the course of the primary MLR, is capable of inducing naive CD8+ T cells to become suppressor effectors but only when these CD8 T cells are in direct contact with allogeneic stimulators. Allospecificity is conferred by the stimulator cells and not by the suppressor-inducer factor. The supernatant of day-5 primary MLR is also capable of inducing antigen-specific suppressor effectors from naive CD8+ T cells, and also only in the presence of allogeneic stimulator cells. Recombinant human IL-2, in doses that are up to five times the amount present in the supernatant cultures, is unable to induce suppressor-effector cells from naive CD8+ T cells. We conclude that, to become allospecific suppressor effectors, naive CD8+ T cells require contact with allogeneic stimulator cells and either CD4+ CD45R+ suppressor inducer cells or suppressor inducer factor/s produced during the course of the primary MLR. PMID- 2572084 TI - [Preoperative ultrasonic diagnosis of maldescended testis]. AB - In 30 boys with a total of 35 maldescended testicles, the present authors found that ultrasonic examination was a useful aid in the diagnosis of maldescended testicles, including intraabdominal testicles. Ultrasonic examination is recommended preoperatively in boys with non-palpable testicles in order to provide optimal surgical intervention. PMID- 2572085 TI - Effects of the alpha 2-adrenoceptor antagonists yohimbine and idazoxan on kidney function in intact and diabetes insipidus rats. AB - The purpose of the present study was to examine the effect of the alpha 2 adrenoceptor antagonists yohimbine and idazoxan on kidney function in intact Sprague-Dawley rats and in Brattleboro rats with familial hypothalamic diabetes insipidus. Both drugs, injected intravenously at 0.5 mg/kg, led to a marked decrease of urine flow, sodium, potassium and chloride excretion in both strains of rats. Glomerular filtration rate and effective renal plasma flow did not change significantly in either strain. The study demonstrates that the alpha 2 adrenoceptor antagonists induce an antidiuretic effect without changes in renal hemodynamics. Antidiuretic hormone does not play a role in yohimbine- or idazoxan induced antidiuresis. The data suggest a role for alpha 2-adrenoceptors in the renal tubular handling of water and salt. PMID- 2572086 TI - Evaluation of monoclonal antibodies to K88, K99, F41 and 987P fimbrial adhesins for the detection of porcine enterotoxigenic Escherichia coli in paraffin-wax tissue sections. AB - A panel of monoclonal antibodies against fimbrial adhesins of porcine enterotoxigenic Escherichia coli were evaluated for the detection of enteric colibacillosis in paraffin-wax embedded sections of piglet small intestine. Using the immunoperoxidase technique, monoclonal antibodies were used to detect epitopes on the K99 adhesin and on the a and c regions of the K88 adhesin. However, monoclonal antibodies to the F41 and 987P adhesins failed to react in sections with organisms colonising the intestine of gnotobiotic piglets monoinfected with strains bearing those adhesins, whereas corresponding polyclonal antisera gave positive results. In contrast to apparent expression of all K99 organisms, only a proportion of organisms were identified by monoclonal or polyclonal antibodies as expressing K88. In some instances, failure of immunostaining was attributed to prolonged storage of tissue in formalin. PMID- 2572087 TI - Segregation of cauliflower mosaic virus symptom genetic determinants. AB - We have created a series of hybrid cauliflower mosaic virus (CaMV) genomes between a severe virus strain (Cabb BJI) and a mild strain (Bari 1) to map the virus genetic loci responsible for specific systemic symptom characters produced in infected turnip plants. Recombinants were generated in vivo by recombinational rescue and in vitro by restriction enzyme fragment exchange. On infection, hybrids induced either parental (wild-type) symptoms or segregated parental characters. Some of the engineered hybrid genomes produced novel symptomatic effects not observed in either of the parental strains whilst others reverted to express parental symptom characters following passaging. Determinants defining differences between the two CaMV strains in respect of four specific symptom characters were delimited to separate genome regions. A locus involved in determining the rate of spread of systemic vein clearing symptoms mapped to a region containing part of gene VII and gene I (nts 109-780). This phenomenon is consistent with the putative involvement of the CaMV gene I product in mediating virus movement within infected plants. Determinants influencing the degree of leaf chlorosis were located in a separate genome domain encompassing part of gene VI together with the large intergenic region and part of gene VII (nts 6103-90). Determinants controlling timing of initial systemic symptom appearance were mapped to a region between nts 2150 and 4438 containing part of gene III, gene IV, and part of gene V. Plant stunting was influenced by loci in at least two separate regions, one containing parts of gene I and II, and a second within the reverse transcriptase gene (V). We conclude that symptoms produced by CaMV infection can be subdivided into individual characters, the genetic determinants of which segregate to different virus genetic loci and are not restricted to a single gene product. PMID- 2572088 TI - [Stomach and duodenal ulcer in endocrine tumors]. AB - Clinico-anatomical analysis was performed in 13 cases of endocrine tumors of various sites with concomitant gastric and duodenal ulcer (Zollinger-Ellison's syndrome). Ulcer tended to recur in these patients; it was associated with pronounced pain and involved a high rate of such complications as perforation, bleeding and gastrointestinal fistula. Endocrine tumors were found to locate in the pancreas (5) and duodenum (5) while, in the other three patients, multiple endocrine adenomatosis (type 1) was identified. Correct diagnosis was made in three cases only and surgery was successful in two of them. Errors are analysed. PMID- 2572089 TI - [Genetic study in 8 families with a child with cystic fibrosis]. AB - Cystic Fibrosis (CF) is the most common genetically determined disease in Caucasians. Major advances in our understanding of the genetics of cystic fibrosis have occurred during the past couple of years. The CF gene has been localized at 7 q31 by the use of linkage analysis with protein and DNA polymorphisms. DNA probes showing close linkage to the CF gene have been discovered and are being used to identify the gene itself and also for purposes of genetic diagnosis. We investigated 35 blood samples from members of 8 CF families with the probes J3.11/MspI, metH/TaqI, metH/MspI, metD/TaqI and 7 c22/EcoRI. In 8 out of 11 siblings of our CF patients we got full information about the carrier status. Since we investigated the siblings as well as the parents of our patients, these results may be of benefit for a later prenatal diagnosis. PMID- 2572090 TI - Do all histamine2-antagonists cause a warfarin drug interaction? AB - Cimetidine, the first marketed histamine2-receptor antagonist, has been shown to decrease the clearance of warfarin consistently through inhibition of cytochrome P-450 metabolism. The clinical significance of this drug-drug interaction has been questioned due to: (1) the lowering of the warfarin therapeutic range, (2) the lowering of the total daily therapeutic cimetidine dosage, (3) the advent of once-daily cimetidine dosing, and (4) the demonstration that the clearance of the less active warfarin R-enantiomer is decreased to a greater extent than the more active S-enantiomer. Ranitidine has been implicated in both increasing and decreasing warfarin's hypoprothrombinemic-effect (noted in the warfarin package insert), despite the majority of investigations demonstrating no warfarin clearance changes. Careful examination of the implicating data indicates that the majority of the warfarin pharmacodynamic and pharmacokinetic variance that occurs with combined ranitidine-warfarin therapy cannot be attributed to a drug-drug interaction. No data are available to implicate the newer histamine2-antagonists, famotidine and nizatidine, in causing a decrease in warfarin metabolism. PMID- 2572091 TI - Neuroleptic malignant syndrome: successful rechallenge with thioridazine. PMID- 2572093 TI - [Processing of a speech signal and thereby necessary attention: a contribution to dual performance]. AB - Speech intelligibility during the performance of a second task (sorting of small plates), and the frequency of sorting in dependence of the phases of speech processing (input-processing-output) are investigated. A fixed speech level (65 dB) is combined with 5 different noise levels (55, 60, 65, 70, 75 dB). The speech material and the sorting task vary in difficulty (words, sentences, small texts; simple and complicated sorting). By rating 3 questions the subjective quality of both tasks is inquired. MAIN RESULTS: speech intelligibility and frequency of sorting vary in dependence of noise level; frequency of sorting varies in dependence of the phases of speech processing and speech material; subjective ratings are corresponding with the performance of both tasks. PMID- 2572092 TI - [Disorders of attention in school children: recent results of classification, diagnosis and pedagogic-psychologic perspectives for intervention]. AB - Attention disorders in school children are a frequently occurring phenomenon in the work of clinical psychologists and psychiatrists. The new approach (DSM III) emphasizes the role of attention deficit with/without hyperactivity in contrast to the Hyperkinetics syndrome (ICD 9) and offers an alternative interpretation. Based on experimental results and clinical observation, the following characteristic features of this class of disturbances are accentuated. A deficient central activation (probably related to "weakness" of central norepinephrine and the locus for stimulant drug action), an inadequate inhibitory control function, and a (secondary) disturbance of perceptive and logic processing strategies. A method of cognitive-behavioural intervention is offered which may be applied separately of in connection with drug treatment. PMID- 2572094 TI - [Activity status in psychophysiologic studies. "Changes in the general activity level in psychophysiologic studies--an analysis of objective and subjective parameters"]. AB - The general activation level (GAL) is one of the sources of influence in psychophysiological experiments. That means most of the results are dependent on the GAL. Taken this into account one can furthermore elucidate a part of the high variability of the interesting physiological data. We analysed heart rate (HR), auditory evoked potentials (AEP) elicited by background stimuli and the subjective assessment of the internal state of the subjects to describe changes of the GAL. Summarizing the results we can show different influences of the GAL on the physiological data. Changes of HR and AEP-parameters are caused by specific experimental conditions. The distinct influence of the subjective state of the subjects on the AEP-shapes allows the conclusion that subjective data should be more taken into consideration. The comprehensive characterization of the GAL requires data of different integrative levels. PMID- 2572095 TI - [About the question of indication for surgery of retractile testis]. AB - Histometric studies in 34 boys with retractile testes and 21 controls with normal descent showed no significant statistic differences with regard to tubulus diameter and spermatogonia count. Operative treatment of retractile testes, therefore, is not indicated. PMID- 2572096 TI - [Morphological properties and adhesiveness of bacteria of the genus Proteus]. AB - Out of 100 Proteus strains isolated from patients with purulent inflammatory, urological and enteric diseases, from healthy persons and from the environment, 29 stains showed the positive D-mannose-resistant reaction of hemagglutination with chick red blood cells and 18 strains showed such reaction with goose and duck red blood cells. The results of these studies permit the use of chick red blood cells as target cells for the detection of Proteus adhesin. Human red blood cells of groups O, A, B and AB, sheep, bovine, dog, rat and rabbit red blood cells gave no positive D-mannose-sensitive reaction and D-mannose-resistant reaction of hemagglutination. In bacterial cells pili function as organelles which determine Proteus adhesiveness, while flagellae play no positive role. PMID- 2572097 TI - [Effect of nootropic agents on the brain bioelectrical activity and on the indices of neuromediator metabolism in the acute period of severe craniocerebral trauma]. AB - Clinico-biochemical examination and EEG were conducted in 39 patients with severe craniocerebral trauma who were given nootropic agents in a complex of intensive therapy measures. Four types of changes of monoamine metabolism in treatment with piracetam were revealed which were combined with two types of EEG changes. The authors recommend the time for beginning piracetam therapy depending on the level of traumatic injury to the brain. PMID- 2572098 TI - The antifibrillatory effect of fentanyl, sufentanil and carfentanil in the acute phase of local myocardial ischaemia in the dog. AB - To determine the effect of strong analgesics on electrical instability (vulnerability to ventricular fibrillation), we examined the action of fentanyl (60 micrograms/kg), sufentanil (10 micrograms/kg) and carfentanil (3 micrograms/kg) on the ventricular fibrillation threshold (VFT) in a dog model of coronary artery occlusion. The effect of strong analgesics was compared with that of the first-generation beta-blocker propranolol (1.2 mg/kg) and the third generation beta-blocker celiprolol (3 mg/kg). In the 5th minute of ischaemia, VFT declines in all groups of dogs. VFT values rise significantly from the 23rd to the 60th minute of ischaemia after opiate analgesic administration. The mean increase in VFT after opiate analgesics is less pronounced than after beta blockers. Administration of opioid analgesics is followed by strong prevalence of vagal drive to the heart. Sympathetic drive to the heart after propranolol administration disappears and is depressed after the administration of celiprolol only. Blockers of the beta-receptors apparently intervene directly in the ischaemic focus and affect the "substrate" of electrical instability. The opioid analgesics probably exert their effect through a change in "substrate" modulation. There is an ongoing search for drugs that would stabilize the heart electrically in acute myocardial infarction, thus preventing sudden death. Our experimental results favour a combination of strong analgesics with other electrostabilizing drugs. PMID- 2572099 TI - Changes in sodium-potassium ratio in guinea pig epidermis in n-hexadecane-induced hyperplasia. AB - A transient epidermal hyperplasia was induced in guinea pig epidermis by a single application of n-hexadecane. The epidermal response was analysed by light microscopy and by energy dispersive X-ray microanalysis (EDX). The epidermal hyperplasia reached a maximum between 96 and 192 h after the application. The hyperplastic response was associated with a depressed sodium-potassium ratio (increased potassium, decreased sodium) in the keratinocytes at 96 h, beginning already at 48 h. At 24 h there were no major differences in elemental content, compared the controls. The result of the present study is consistent with the hypothesis that alterations in the functional state of the epidermal keratinocytes are associated with changes in the sodium-potassium ratio in the cells. The absence of major elemental changes at 24 h indicates that the initiation of the hyperplastic response occurred prior to this time point. PMID- 2572100 TI - Correlation between urinary melanin-related metabolites and tumour weight in melanoma-bearing mice. AB - Melanoma tissues produce not only 5-S-cysteinyldopa (5-S-CD), a pheomelanin precursor, but also 5,6-dihydroxyindoles, eumelanin precursors. We compared 5-S CD and eumelanin-related metabolites regarding the correlation of their production in melanoma and excretion in urine, versus the weight of B16 mouse melanoma. It was found that B16 melanoma tissues produce 5,6-dihydroxyindole-2 carboxylic acid and its two O-methyl derivatives, the latter of which are partly conjugated with sulphuric acid and then excreted. These indolic metabolites are referred to as total indoles. Both total indoles and 5-S-CD in melanoma and in urine correlated well with each other and reflected the tumour weight. Total indoles had a four-fold greater level of excretion than 5-S-CD, suggesting that the urinary excretion of total indoles is a quantitatively more significant marker of melanoma progression in this melanoma. PMID- 2572101 TI - Expression of interleukin-6-like molecules and tumour necrosis factor after topical treatment of psoriasis with a new vitamin D analogue (MC 903). AB - Skin biopsies from 5 patients with chronic plaque psoriasis were examined to test the effect of topical treatment with a new synthetic vitamin D3 analogue, MC 903, on epidermal interleukin 6 (IL-6) and tumour necrosis factor alpha (TNF alpha). Skin biopsies, taken before therapy and after one and 2 weeks of therapy were examined immunohistologically. IL-6 was visualized using a partially purified polyclonal antiserum to crude supernatants of activated human blood monocytes before and after absorption with recombinant human IL-6. TNF alpha was demonstrated using a specific polyclonal antiserum to human recombinant TNF alpha. Both the intensity and extension of staining for IL-6, but not for TNF alpha, were increased in lesions compared with unaffected skin. During therapy with MC 903 a decline in the staining intensity for IL-6, but not for TNF alpha, was observed in both lesional and unaffected skin. Placebo-treated lesions, however, remained unchanged. PMID- 2572102 TI - Tissue-type plasminogen activator concentrations in plasma from patients with psoriasis. AB - An ELISA for tissue-type plasminogen activator (t-PA) was developed and used to measure the concentration of t-PA in plasma from 15 healthy donors and 23 psoriasis patients. The mean value for the healthy donors was 2.4 +/- 1.3 ng/ml (SD) of t-PA in plasma. The mean value for the psoriasis patients was 4.4 +/- 3.2 ng/ml of t-PA. The psoriasis patients were divided into two groups: patients with mild psoriasis, i.e. less than 20% of the skin affected; and patients with severe psoriasis, i.e. more than 20% of the skin affected. Patients with mild psoriasis had t-PA concentrations of 2.8 +/- 2.3 ng/ml, while patients with severe psoriasis had 5.4 +/- 3.3 ng/ml. t-PA concentrations were higher in the psoriasis patients than in the normal controls (at the 5% level). The patients with severe psoriasis had t-PA concentrations significantly higher than the normal controls (at the 1% level). PMID- 2572103 TI - The effect of cyproterone acetate on hair roots and hair shaft diameter in androgenetic alopecia in females. AB - Twenty female patients suffering from androgenetic alopecia were treated during one year with 50 micrograms ethinyloestradiol plus 2 mg cyproterone acetate and additional 20 mg cyproterone acetate on days 5-20 of the menstrual cycle. The control group consisted of eight untreated female patients with androgenetic alopecia. The parameters used to evaluate therapeutic results were trichogram, hair shaft diameter of full anagen and number of hairs measuring less than 40 microns. Hair roots were epilated from two locations of the scalp: fronto-cranial and left temporal (reference point). The trichogram of the fronto-cranial scalp region showed an increase of anagens as well as a decrease of telogens. These changes were statistically highly significant. Further, there was a decrease of dysplastic/dystrophic forms. The left temporal scalp region showed no significant differences. The mean hair shaft diameter of full anagen (n = 8) increased, while the number of hairs measuring less than 40 microns (n = 8) decreased. The last two findings showed no statistically significant differences. The therapeutic results warrant the conclusion that cyproterone acetate seems to be effective in androgenetic alopecia in women. PMID- 2572104 TI - Monoclonal antibodies to collagens for immunofluorescent examination of human skin. AB - Monoclonal antibodies, specific for each of human types I, III, IV and V collagens, were produced and shown to be suitable for immunofluorescent studies of human skin. The antibodies showed that types I and III collagens were abundant and distributed throughout the dermis. The distribution of type III collagen appeared different in the papillary compared with the reticular dermis, although an increased concentration of type III collagen in the papillary dermis could not be unambiguously established. Type V collagen, which could be visualised only after acid-pretreatment of sections, was also distributed throughout the dermis, but appeared to be in higher concentrations around cells. Type IV collagen was observed specifically in the basement membrane associated with the dermal/epidermal junction. PMID- 2572105 TI - Expression of CD36 (OKM5) antigen on epidermal cells in normal and diseased skin. AB - The expression of the CD36 (OKM5) antigen was studied with the PAP technique on sections of skin from healthy subjects and of normal and diseased skin from patients with various skin diseases. In specimens from healthy subjects the antigen was found on vascular and perivascular structures and in some cases it was seen on cells of the acrosyringium. A net-like pattern of CD36 was observed in the upper part of the stratum spinosum in skin lesions of patients with psoriasis, lichen planus, pityriasis rosea, morbilliform drug reactions, necrobiosis lipoidica, lichen amyloidosus, Darier's disease and ichthyosis vulgaris. Expression of CD36 was also seen in the nonlesional skin just below the granular layer in 3 of 5 patients with factitial urticaria with immediate dermographism, but not in delayed dermographism or chronic urticaria. In ichthyosis vulgaris CD36 was also expressed in dendritic cells of the basal layer and in a patient with a graft versus host reaction it was recognized both on scattered keratinocytes and on dendritic cells in the epidermis. The role of the expression of the CD36 antigen in the skin is unknown. The activated cells might possibly serve as antigen-presenting cells and/or have a modulatory influence on an inflammatory reaction. PMID- 2572106 TI - Circular and branched Birbeck granules and cytomembrane blebbing in Langerhans' cells after dithranol irritation. AB - Mild dithranol irritation of healthy human skin has a stronger effect on the fine structure of Langerhans' cells (LC) than on that of other epidermal cells, causing mitochondrial enlargement and disruption of the cristae of LCs. With a stronger dithranol irritation, LCs were even more affected resulting in circular and branched Birbeck granules (BG) and blebbing of the LC cytomembrane. More often than is normal, BGs were contiguous with the LC cytomembrane. Electron microscopic observations indicated that blebbing and the abnormal BG formation were associated phenomena, in accordance with the hypothesis that BGs are endocytotic organelles formed from the cytomembrane. PMID- 2572108 TI - Chronic recurrent annular neutrophilic dermatosis. An entity? AB - Two cases with recurrent annular lesions of similar clinical appearance and course are described. The histopathological investigations showed in both cases dense and diffuse infiltrates mainly of polymorphonuclear neutrophilic leucocytes in the mid dermis. No fever, leucocytosis or elevated ESR were observed. Systemic corticosteroid treatment controlled symptoms but recurrence after treatment was withdrawn occurred in both cases. It is discussed if we are dealing with a variant of Sweet's syndrome or a new entity. PMID- 2572107 TI - Computerized recording of itch in patients on maintenance hemodialysis. AB - Itch was assessed both continuously using a computerized method, Pain-Track, and retrospectively using visual analogue scales (VAS) by 28 patients undergoing maintenance hemodialysis and suffering from uremic pruritus. Measurements were performed during 7 consecutive days including three dialysis sessions. Pain-Track recordings showed that itch intensity was greater during dialysis than on days following dialysis (p less than 0.05). Possible explanations are that pruritogenic substances might be released during treatment or that removal of such substances during dialysis leads to amelioration of symptoms after treatment. Alternatively, lowering of the sensory threshold due to general discomfort in association with dialysis may exacerbate the itch intensity. There was no consistent difference between daytime and bedtime itch scores over the week, except on the second day without treatment, when bedtime itch ratings significantly exceeded those during the day (p less than 0.05), suggesting that factors other than inactivity are essential for this peak in itch intensity. Thus, after 2 days without treatment, when patients become increasingly metabolically deranged, they reported maximal itch, implying that the accumulation of pruritogenic substances is of major importance in the pathogenesis of uremic pruritus. There was a positive correlation between Pain Track and VAS data, although significant fluctuations in itching could be detected only with Pain-Track. PMID- 2572109 TI - An epidemiological study of rosacea. AB - In a non-selected population of 809 office employees (454 women and 355 men) 81 persons were diagnosed as having rosacea, giving a prevalence of 10% (women 14%, men 5%). The rosacea group was compared with the rest of the study population. Most of the cases were rather mild. The rosacea was of an erythematotelangiectatic type in 81% of the cases and of a papulopustular type in 19%. Unilateral lesions were found in 11 subjects (14%). Only 17% of those with rosacea were impaired by sunlight, whereas 26% improved. In the rosacea group, 27% were found to suffer from migraine and 42% from a tendency to flush, compared with 13% (p less than 0.001) and 16% (p less than 0.001) respectively in the comparison group. Flushing and the regulatory mechanism of the blood vessels thus seem to be of importance in the pathogenesis of rosacea. Individuals with good pigmentation ability showed a tendency to a decreased occurrence of rosacea. The frequency of eye complaints was the same in the two groups. PMID- 2572110 TI - Local involvement of antigen-presenting cells and activated T cells in perilesional and clinically uninvolved skin in pemphigus vulgaris. AB - Biopsies obtained from both the perilesional areas and clinically uninvolved skin of patients with pemphigus vulgaris (PV) were studied for antigen-presenting cell and lymphocyte phenotype and/or activation phenotype using monoclonal antibodies in avidin-biotin-peroxidase complex staining. Perilesional PV skin contained CD4+ and CD8+ T lymphocytes as the predominant cell type, but cells with a potential antigen-presenting function displaying CD11b phenotype of monocyte/macrophages and, in particular, CD1 phenotype of Langerhans cells were also present. The number of mononuclear inflammatory cells was greater in perilesional than in clinically uninvolved PV skin, and so were the proportions of CD4+, CD8+, CD25+, Ia+ cells (p less than 0.01), and CD1+ Langerhans cells and transferrin receptor positive cells (p less than 0.05). These findings confirm and extend earlier observations on local involvement of immunocompetent cells in PV. PMID- 2572112 TI - Skin extensibility time in women. Changes in relation to sex hormones. AB - The influence of female sex hormones on mechanical properties of the skin has been assessed in an in vivo extensometric study. Twenty young (20 +/- 4 years) and 12 middle-aged healthy women (42 +/- 3 years) entered the study. Measurements were carried out on the volar surface of the left forearm on the 10th and 25th day of the menstrual cycle. A significantly decreased skin extensibility time in the pre-menstrual phase was found (25th day) when compared with the 10th day in the young group, while the older one did not reveal significant changes. The data are compatible with an increased water content of the skin noticeable in the pre menstrual phase and more relevant in young women. In studies on mechanical properties of the skin, changes relative to sex hormones and menstrual cycle need to be taken into account. PMID- 2572111 TI - Human keratinocytes in vitro have receptors for leukotriene B4. AB - We have demonstrated a high-affinity binding site for leukotriene B4 (LTB4) on human epidermal keratinocytes in vitro. In substrate saturation studies, one population of binding sites with a dissociation constant (Kd) of 1.03 +/- 0.3 nM and a maximal binding capacity (Bmax) of 148.2 +/- 45.3 fmol/mg protein could be demonstrated. On average 5,500 binding sites were found on individual keratinocytes in culture. The affinity constant of this binding site correlates well with previous reports on the proliferative effect of LTB4 on keratinocytes in vitro. These findings confirm that LTB4 may in part be responsible for epidermal hyperproliferation in inflammatory skin diseases. PMID- 2572113 TI - Evaluation of the effect of heparin and tetracycline on the cohesion of the dermal-epidermal junction. AB - In an attempt to demonstrate the effect of tetracycline or heparin therapy on the dermo-epidermal junction, the suction blister time was measured at the beginning and at the end of the study in 18 female Sprague-Dawley hairless rats, using the suction blister method. Suction blister time was defined as the time taken for the first sign of a vesicle to appear in the hole in the diaphragm of a suction chamber. 200 IU heparinate calcium was administered daily for 6 days in 6 rats. Six other rats received 27.5 mg tetracycline chlorhydrate daily for 6 days. The others did not receive any drug. Significant statistical differences in suction blister time before and after administration of the drugs were found in the heparin group (p less than 0.05), and in the tetracycline group (p less than 0.05), compared with the control group. Heparin and tetracycline were found to increase suction blister time significantly. These results suggest an increased cohesion in the dermo-epidermal junction due to heparin and tetracycline. PMID- 2572114 TI - Human epidermal Langerhans' cells are sensitive to rapid cooling by ethyl chloride. AB - Topical anesthesia with ethyl chloride spraying generates some remarkable reactive events in the human Langerhans' cell (LC) system. Many LC retract their dendrites and a considerable number move to the innermost layers of the epidermis within 15 minutes. This rapid motility supports the view that the interstices between cells in living epidermis are large. The LC cytomembrane is very susceptible to cold shock which causes the cytomembrane to superimpose upon itself forming abnormally shaped Birbeck granules. This process may consume too much of the cytomembrane to be compatible with cell survival. PMID- 2572115 TI - Etretinate reduces connective tissue degeneration in lichen sclerosus et atrophicus. AB - Retinoids have effects on the metabolism of keratinization and on the metabolism of connective tissue. Recent results have indicated that they may be helpful for treating dermatological diseases which involve marked connective tissue changes such as scleroderma, keloids and actinic skin damage. In addition, retinoids have been shown to reduce the clinical and histological alterations occurring in vulvar lichen sclerosus. For these reasons, etretinate was tried in a patient with extensive lichen sclerosus et atrophicus (LSA). Clinical improvement was seen after three months' treatment, i.e. a decrease in pruritus and softening of the skin. The degenerated zone in the lesional skin was shown by histological analyses to have reduced markedly. The immunohistochemistry, with unaltered staining for type III procollagen and fibronectin, disclosed no signs of enhanced collagen synthesis. Thus the reparation mechanism remained obscure. PMID- 2572116 TI - Lipodystrophia centrifugalis sacralis infantilis. A 15-year follow-up observation. AB - A unique case of Lipodystrophia centrifugalis sacralis infantilis in a caucasian is reported. This case fulfils all the clinical requirements of the centrifugalis lipodystrophy described in Oriental children by Imamura et al. (1) and usually localized on the abdomen. The 15-year follow-up of this case clearly demonstrates the tendency toward spontaneous remission of the disease after puberty. PMID- 2572117 TI - Sweet's syndrome associated with Crohn's disease. AB - The first known case involving an association of Sweet's syndrome with Crohn's disease is described. A 36-year-old woman developed a diarrhea, fever, and infiltrated erythematous cutaneous plaques on neck and limbs, consistent with a presumptive diagnosis of Sweet's syndrome. This was confirmed by a skin biopsy showing a dense dermal infiltrate of polymorphonuclear leukocytes. Crohn's disease, extending from the anus to the terminal ileum, was diagnosed as well. Prednisolone treatment resulted in the improvement of both the bowel disease and skin lesions. PMID- 2572118 TI - Transient acantholytic dermatosis associated with lymphomatous angioimmunoblastic lymphadenopathy. AB - Transient acantholytic dermatosis is a papulovesicular cutaneous eruption first described in 1970. There have been subsequent reports of similar disorders occurring in patients with malignancy. Angioimmunoblastic lymphadenopathy with dysproteinemia is a disorder characterized by an acute onset of generalized lymphadenopathy associated with fever, malaise, pruritus, night sweats, and hepatosplenomegaly. The patient described had a papular acantholytic dermatosis associated with the development of angioimmunoblastic lymphadenopathy with dysproteinemia-like T-cell lymphoma. The cutaneous manifestations of angioimmunoblastic lymphadenopathy with dysproteinemia are discussed. PMID- 2572120 TI - The use of a corticosteroid cream for immediate reduction in the clinical signs of acne vulgaris. AB - A controlled trial of the anti-inflammatory effect of a steroid cream (clobetasol propionate) in 11 patients with moderate acne was assessed over a 3 week period. The placebo cream was the base of the steroid cream. Therapeutic effect was assessed by the number of active and less actively inflamed papules and pustules present at the beginning and end of the 3 week trial period. No significant change in lesion counts was observed. This result indicates that a potent topical steroid cream produces no short-term improvement in patients with moderate acne. PMID- 2572119 TI - Outpatient treatment with short-contact dithranol. The impact of frequent concentration adjustments. AB - Short-contact therapy with dithranol has become an important possibility for the treatment of psoriasis on an outpatient basis. Two groups of patients with chronic stable plaque psoriasis were treated on an outpatient basis with a short contact regime using dithranol in a stiffened paraffin base. In one group the concentrations were adjusted once a week and in the other group 3 times a week. Clinical results were evaluated using the Psoriasis Area Severity Index. In the group of patients which was seen 3 times a week, the improvement was significantly better. The duration of the treatment period was significantly shorter compared to the group seen once a week due to the shorter time interval in which the optimal concentration of dithranol was reached. In addition, the incidence and severity of irritation was more pronounced in the latter group. PMID- 2572121 TI - Wound dressing after skin planing. AB - A variety of dressings have been used following skin planing. Compresses are difficult to apply and simple bandages stick tight and are hard to remove. The Department of Dermatology has for several years used Collagen film, but this film is no longer commercially available. After 12 patients with acne scars underwent skin planing, synthetic polyurethanefilm (Omiderm) was applied for 7 days, with good results. PMID- 2572122 TI - Cervical signs of HPV infection in PAP-smear negative women with external genital warts. AB - Forty-eight women with external genital warts, all with normal cervical cytological PAP smears, were examined by means of colposcopy. One cervical biopsy for histological evaluation was taken from each woman, irrespective of the colposcopic findings. Koilocytosis was detected in 18/48 (38%) and dysplasia (CIN 1) in 3/48 (6%) of the patients. The presence of aceto-white lesions on the cervix was significantly associated with abnormal histology; 12 of 17 (71%) aceto white lesions and 8 to 31 (26%) normal-appearing cervices showed histological changes indicating HPV infection (p less than 0.01). Women with koilocytosis and dysplasia had genital warts for a mean of 201 days compared with 79 days in women with normal cervical histology (p less than 0.01). It is concluded that even the clinically normal appearing cervix frequently is a reservoir for HPV and that colposcopy should be a routine procedure in women with external genital warts, irrespective of the result of the PAP smear, to provide a basis for proper counselling and individual therapy. PMID- 2572123 TI - Effects of repeated application of a moisturizer. AB - Epidermal hydration following repeated application of an oil in water emulsion was studied on the forearm skin of 16 healthy females by non-invasive methods. The lotion was applied twice daily for 7 days, and values were followed 7 days after cessation of treatment. The opposite forearm served as an untreated control. Electrical conductance and capacitance showed similar results, i.e. increased values (p less than 0.001) after 2 days of application, reaching a plateau during further applications. Two days after cessation, values were still increased (p less than 0.001), and the conductance was also increased 7 days after cessation of treatment. The water evaporation and the cutaneous blood flow did not change, i.e. indicating no mild irritant effect. Skin surface lipids did not change, i.e. indicating that no significant amounts of emulsion oil remained on the skin at the time of recording. Probably components of the oil phase of the emulsion are absorbed into the epidermis, which is associated with improved hydration as a later event. PMID- 2572124 TI - Impaired somatostatin response to orally administered glucose in type II diabetes entails both somatostatin-28 and -14 and is associated with deranged metabolic control. AB - We have investigated the effects of hyperglycemia in Type II diabetic patients on the somatostatin response to oral glucose. In these patients hyperglycemia prevailed (11.8 +/- 1.4 mmol/l) and was markedly increased to a maximum of 18.9 +/- 1.0 mmol/l following the ingestion of 75 g of glucose. The rise in blood glucose following glucose ingestion failed to induce a rise in plasma levels of somatostatin-like immunoreactivity. Biostator-regulated insulin infusion normalized fasting levels of blood glucose and reduced the hyperglycemia following glucose ingestion, i.e. blood glucose now rose from 4.6 +/- 0.1 to a maximum of 7.3 +/- 0.8 mmol/l. This moderate rise in blood glucose was accompanied by a significant (p less than 0.05) rise in somatostatin-like immunoreactivity. Somatostatin-28 and somatostatin-14 were separated using a Sephadex G-50 fine column. Biostator treatment suppressed plasma levels of both peptides during fasting conditions. Treatment was also accompanied by a rise in both peptides during the first hour following glucose ingestion; this rise did not occur in the untreated state. IN CONCLUSION: lack of somatostatin response to glucose in non-insulin-dependent diabetes mellitus is associated with deranged metabolic control. Unresponsiveness to glucose entails the secretion of both somatostatin-28 and -14. PMID- 2572125 TI - Hypersensitivity to arginine of both B and D pancreatic cells in adult streptozotocin-diabetic rats. AB - Insulin and pancreatic somatostatin secretions were studied after stimulation with an arginine infusion (5 mmol/l) in isolated perfused pancreata of adult streptozotocin-diabetic rats. In the presence of 2.8 mmol/l glucose, arginine clearly stimulated insulin and somatostatin secretions in diabetic rats, whereas it was ineffective in normal rats. Thus, not only the B-cells, but also the D cells of the pancreas from streptozotocin-diabetic rats are hypersensitive to arginine. The infusion of insulin (4 U/l) did not modify this hypersensitivity of the D-cells to arginine in pancreata of streptozotocin-diabetic rats. PMID- 2572127 TI - The stimulated C-kinase activity in estradiol-treated rat pituitaries is reduced by chronic treatment with the dopamine agonist CV 205-502. AB - To investigate whether the modulation of lactotrope cell multiplication and prolactin secretion in rat pituitary glands implicated the phosphoinositide C kinase system, female Wistar rats were treated or not-with the dopamine agonist CV 205-502 or 8 days or with estradiol cervical implants for 8 for 15 days, alone or in combination with CV 205-502 for the last 8 days. CV 205-502 treatment induced a significant reduction in plasma PRL levels and in pituitary weights, whereas estradiol treatment induced a significant increase in both parameters. CV 205-502, in association with estradiol, counteracted estradiol stimulation of PRL levels and of pituitary weights. Total C-kinase activity in controls was 29.8 +/- 9.9 pmol 32phosphorus/min (N = 7, mean +/- SEM), mainly found in the soluble fraction (84%). When administered alone, CV 205-502 induced a significant reduction (-58%, p less than 0.02) in C-kinase activity in the particulate fraction with no modification in the soluble fraction. Both 8 and 15 days estradiol treatment induced a significant stimulation of total C-kinase activity, 74% and 155% respectively. When combined with estradiol, CV 205-502 significantly (p less than 0.02) counteracted the estradiol increase in total C-kinase activity, which was only 45% over control values. We conclude that treatment with a dopamine agonist and estradiol, which have antagonistic effects on the pituitary, exerts an opposite regulation of C-kinase activity. Whether this may be one of the mechanisms involved in their interaction on pituitary lactotropes remains to be determined. PMID- 2572126 TI - Bovine neurophysin-II stimulates prolactin release without involvement of dopaminergic prolactin-release inhibiting factor receptor in the estradiol-primed male rat. AB - Neurophysins have been considered to be physiologically inert carrier proteins for the neurohypophysial hormones, oxytocin and vasopressin. We have observed that bovine neurophysin-II indirectly stimulates prolactin release in estradiol primed male rats. The release of prolactin is regulated by a dual hypothalamic control system, the prolactin-release-inhibiting factor and the prolactin releasing factor. We have tried to clarify whether neurophysin-II is acting through stimulation of prolactin-releasing factor by eliminating the possibility of dopaminergic prolactin release-inhibiting factor release. Male rats were primed with estradiol and functional dopaminergic prolactin release-inhibiting factor receptors were completely blocked by pretreatment with a large dose of pimozide (3 mg/kg), a dopaminergic receptor blocking agent. The neurophysin-II stimulated prolactin release in the rats which did not have any functional dopaminergic prolactin release-inhibiting factor receptors suggesting that neurophysin-II likely initiates a chain of events which eventually stimulates prolactin-releasing factor release since the possibility of involvement of the dopaminergic prolactin release-inhibiting factor system is eliminated. Opioids are known to be one of a chain of events which transmit external stress into a stimulation of prolactin release. Naloxone, a mu-receptor antagonist, was injected 20 min before neurophysin-II administration into rats which were primed with estradiol and pretreated with pimozide (3 mg/kg), but the naloxone administration did not block the prolactin release stimulated by neurophysin-II injection. This result indicates that opioids are not one of the chain of events between initiation of stimulation by neurophysin-II and prolactin release. PMID- 2572128 TI - Characterization of alpha 2-adrenoceptors in a plasma membrane enriched fraction from the insulin-secreting cell line RINm5F. AB - A plasma membrane enriched fraction from the insulin-secreting cell line RINm5F was used to characterize [3H]clonidine binding. After a single self-generating Percoll gradient, the specific activity of 5'-nucleotidase (a plasma membrane marker) of the membrane fraction was enriched about 8-fold over that of the homogenate and nearly 30% of the total amount was recovered. The fraction was essentially free of mitochondria and secretory granules. [3H]clonidine binding to this membrane fraction revealed a single, high affinity binding site with a Kd of 2.3 nmol/l. The binding was competitively inhibited by adrenergic agonists in the following order of potency: clonidine greater than epinephrine greater than phenylephrine greater than isoproterenol, and by antagonists in the order of potency: idazoxan greater than yohimbine greater than propranolol greater than prazosin. Pertussis toxin pretreatment of the cells did not alter the inhibition of [3H]clonidine binding by epinephrine and clonidine nor the estimated receptor number for [3H]clonidine. In conclusion, the pharmacologic characteristics of [3H]clonidine binding sites on a plasma membrane enriched fraction from insulin secreting RINm5F cells demonstrate that the receptor is of the alpha 2-adrenergic subtype. PMID- 2572129 TI - Does sodium valproate increase clinical effects of diazepam? Double blind study. PMID- 2572131 TI - Clinical predictors of schizophrenic patient response to neuroleptics. PMID- 2572130 TI - The immunomodulators levamisole, muramyl dipeptide and adamantylamide dipeptide antagonize the antianxiety effect of diazepam. PMID- 2572132 TI - Incisive neuroleptic drugs in child psychiatry. PMID- 2572133 TI - Isofloxythepine in an open study. PMID- 2572134 TI - Cortisol deficiency in metomidate anesthetized bacteremic pigs: results in circulatory failure--beneficial effect of cortisol substitution. AB - Etomidate and the closely related metomidate are known to inhibit cortisol synthesis. We studied the influence of metomidate on hemodynamic performance and survival time of bacteremic pigs. Thirty pigs, 30.2 +/- 0.8 kg, were anesthetized with intravenous metomidate (2.5 mg.kg-1.h-1) plus ketamine (3.0 mg.kg-1.h-1), and were then mechanically ventilated. The animals were randomly allocated to three groups of 10 pigs each. Group A received an infusion of live Pseudomonas aeruginosa bacteria (2.5.10(9).kg-1.h-1 organisms until death), Group B additionally received a bolus of 1 mg.kg-1 cortisol (followed by an infusion of 0.1 mg.kg-1.h-1) starting 1 h prior to the bacterial infusion, and Group C served as anesthesia control without receiving bacteria or cortisol. The experiments in Group C were terminated after 10 h. In Group A the cortisol level was severely suppressed from the very beginning. The animals died of circulatory failure after 4.3 +/- 0.4 h. In contrast, Group B exhibited fairly stable hemodynamics, but the animals died due to pulmonary edema after 11.1 +/- 1.3 h. Cortisol deficiency in metomidate anesthetized pigs facilitates the development of circulatory failure in the course of Pseudomonas bacteremia, which does not occur if cortisol is infused to reconstitute a physiological level. However, this cortisol substitution did not prevent the development of pulmonary edema caused by Pseudomonas aeruginosa. Possible mechanisms of the deleterious effect of cortisol deficiency and implications in regard to the clinical use of metomidate/etomidate are discussed. PMID- 2572135 TI - Reversal of profound paralysis: use of large doses of edrophonium to antagonize vecuronium and pancuronium induced neuromuscular blockade. AB - The ability to evoke reversal of dense vecuronium- and pancuronium-induced paralysis (T1 10% of control) with edrophonium 1.0 mg.kg-1 was studied using train-of-four nerve stimulation and electromyographic monitoring. Two different end-points, train-of-four ratios of 0.5 and 0.7, were used to define "adequate reversal", and the results for both relaxants were compared. Reversal was reliable and rapid for vecuronium if either ratio was used with times of 2.8 (1.5) and 9 (3) min required to achieve ratios of 0.5 and 0.7, respectively. However, if the block was due to pancuronium, reversal was unreliable with 2 of 9 and 4 of 9 patients not achieving ratios of 0.5 and 0.7, respectively. Reversal was also markedly prolonged in this group with a mean time of 37 (23) min to achieve a ratio of 0.7, and in almost half these patients a supplementary dose of edrophonium was required. PMID- 2572136 TI - The cardiovascular effects of vecuronium during halothane anaesthesia. AB - In a double-blind, placebo-controlled, prospective randomized trial in 70 otherwise healthy patients undergoing elective surgery of the thyroid gland, the breast, the inguinal canal or the lower limbs, we studied the cardiovascular effects of vecuronium during halothane-nitrous oxide anaesthesia. Vecuronium was found to have no clinically significant influence on heart rate or mean arterial pressure, before or during surgery. Bradycardia was a rare event under the special conditions of the study design. PMID- 2572137 TI - A contribution to the monitoring of neuromuscular blockade: an evaluation of the Datex Relaxograph. AB - The Datex Relaxograph is a neuromuscular transmission monitor, which measures the degree of neuromuscular blockade during anesthesia. In order to evaluate the reliability of the Relaxograph, results, obtained with this apparatus, were compared with simultaneous mechanical measurements obtained with the Myograph 2000 (Biometer). Although there was a good correlation between the two throughout the study, a shift towards mechanical responses was observed in all cases. Subsequently, the Relaxograph was used to evaluate atracurium and vecuronium when given accordingly to the priming principle. Results show that neither drug offers major clinical advantages over the other. In a third study, a new bolus-constant infusion regimen of atracurium was evaluated. Results show that it produces a predictable and stable neuromuscular blockade. PMID- 2572138 TI - Differential diagnostic value and influence on EEG of Anexate (Flumazenil, Ro 15 1788) in benzodiazepine-intoxication. A case report. AB - The authors report on a patient, admitted to a regional hospital in a comatose state of uncertain origin. Anexate (Flumazenil, Ro 15-1788) was used to demonstrate the etiology of this benzodiazepine-induced coma. The almost immediate effect of flumazenil injection on EEG in benzodiazepine induced coma is shown. PMID- 2572139 TI - Characterization of the course of senile dementia of the Alzheimer type using cerebrospinal fluid levels of acetylcholinesterase and somatostatin. AB - Through an epidemiological survey, we observed 3 types of clinical courses among patients with senile dementia of the Alzheimer type (SDAT). The mental ability of the patients declined rapidly (Group A; n = 11), gradually (Group B; n = 6), or showed extremely slow changes (Group C; n = 9). The acetylcholinesterase (AChE) activity and somatostatin (SRIF) concentration of the cerebrospinal fluid (CSF) were measured in patients with Alzheimer's disease (AD) and 3 types of SDAT. Both AChE activity and SRIF concentration of CSF were significantly lower in Group A and among patients with AD compared with age-matched control subjects. Both AChE activity and SRIF concentration of CSF were not significantly different in Groups B and C. This biochemical study confirmed our epidemiological finding that only the patients in Group A with SDAT closely resembled the clinical course of AD and may belong to the category of neurodegenerative disorders. PMID- 2572141 TI - Molecular analysis of hypoxanthine-guanine phosphoribosyltransferase mutations in five unrelated Japanese patients. AB - The isoenzyme of hypoxanthine-guanine phosphoribosyltransferase (HPRT, E.C.2.4.2.8) functions in the metabolic salvage of purines. Partial HPRT deficiency is associated with gouty arthritis, while absence of activity results in Lesch-Nyhan (LN) syndrome. We characterized five unrelated patients with HPRT deficiency to understand the spectrum of molecular defects using Southern and Northern blot, polymerase chain amplification of HPRT mRNA and DNA sequencing, and oligonucleotide hybridization analysis of the HPRT gene. Southern blot analysis of DNA indicated that mutations leading to HPRT deficiency in our five patients were not the result of major chromosomal rearrangements or deletions. Sequencing analysis of the amplified DNA from three different patients with HPRT deficiency implied three unique molecular abnormalities: 1) one single-base substitution at codon 54 (from ATG to CTG) resulting in the replacement of methionine with leucine in an LN patient, 2) two single-base substitutions at codon 179 (from GTT to GGT) and at codon 180 (from GGA to AGA) resulting in the replacement of valine with glycine and glycine with arginine in a gouty patient, and 3) 51 nucleotide deletion between nucleotides 747 and 797 resulting in the formation of shorter sized HPRT mRNA and putative two amino-acid deleted HPRT protein in another gouty patient. These results are the direct molecular evidence of genetic heterogeneity in mutant HPRT. PMID- 2572140 TI - [Evaluation of cetirizine and of terfenadine in the treatment of perennial allergic rhinitis]. AB - Thirty patients suffering from perennial allergic rhinitis took part in a cross over, double-blind study during which they were treated for 3 two-week periods with either 10 mg cetirizine nocte or 60 mg terfenadine given morning and evening, or with a placebo. Both the active products were found to significantly improve the symptoms when compared to placebo (p less than or equal to 0.001). Cetirizine helped to reduce the symptoms of a blocked nose to a significantly greater extent than terfenadine (p less than or equal to 0.05). A significantly larger number of patients (p less than or equal to 0.05) preferred cetirizine (17/30) to terfenadine (6/30). Four patients taking cetirizine, and six of those taking terfenadine, reported a mild sedative effect. PMID- 2572142 TI - [Effects of diazepam on the rat open field behavior and high K+ induced release of monoamine neurotransmitters from brain slices]. AB - Acute i. p injection of diazepam (3 mg/kg) significantly decreased the open field behaviors (locomotion and rearing) of rats. However, when diazepam (3 mg/kg) was given ip daily for 15 consecutive days, no depressive effect on the open field behaviors was observed. After the open field observation, rats were immediately decapitated and brain slices of cortex, hippocampus, amygdala and brain stem were stimulated by high K+ (65 mmol/L) to cause the release of noradrenaline (NA), dopamine (DA) and serotonin (5-HT). Diazepam (9 mumol/Ls) significantly depressed the release of monoamine neurotransmitters caused by high K+, which were antagonized by Ro 15-1788 (5 mumol/L) in vitro. Acute administration of diazepam (ip 3 mg/kg) also significantly depressed the release of monoamine neurotransmitters caused by high K+ from all regions observed. However, hippocampus showed tolerance to the depressive effects of diazepam on the releases of NA and DA, and amygdala only on the release of DA, caused by high K+, when given ip chronically (3 mg/kg, 15 d). Our results suggest that the depressive effect of benzodiazepines on the release caused by anxiogenic factors of catecholamine in the hippocampus, front cortex and amygdala regions may be one of the mechanisms of sedative and anxiolytic actions of benzodiazepines. PMID- 2572143 TI - Amphetamine stereotypy in cats and neurotransmitter interactions in the caudate nucleus. II. Effects of intracaudate injection of leu-enkephalin and naloxone. AB - In cats, injection of leu-enkephalin (LEU-ENK) and naloxone (NAL) into the rostrodorsal part of the caudate nucleus 30 minutes after amphetamine i. p. induced quantitative and qualitative changes in amphetamine stereotypy. LEU-ENK decreased the amplitude or the number of the focused stereotyped movements and increased the ipsilateral at the expense of the contralateral head turnings. NAL increased the duration of the phase with head movements, the number of circlings and the number of other locomotor behaviors, as well as the number of the contralateral head turnings at the expense of the ipsilateral ones. LEU-ENK and NAL injected alone (without pre-amphetamine) had not any effects on the animal behavior. The results suggest a role for enkephalins and possibly for other naloxone-sensitive opiates in the behavioural responses to amphetamine in cats. PMID- 2572144 TI - Effects of sympathetic stimulation on C-fibre responses in rabbit. AB - Unmyelinated C-fibre responses to electrical stimulation were recorded in common peroneal, sural and tibial nerves of rabbits. Three distinct C elevations, here called C1, C2 and C3, were recorded. C2 is probably of somatic origin because it was depressed due to collision by peripheral stimulation of cutaneous receptors. The conduction velocity of C3 corresponded to that of sympathetic post-ganglionic fibres. During sympathetic trunk stimulation the A-fibre responses were not significantly changed while C responses, especially C2, were reduced in amplitude and slightly delayed. The C-fibre responses were also influenced by intra arterial infusion of noradrenaline. In most cases, the latency of the response was increased. The effect of sympathetic stimulation was completely blocked by hexamethonium, and partly blocked by phentolamine, an adrenergic alpha-receptor blocking agent which also blocked the effect of noradrenaline. The findings suggest that there are adrenergic receptors distributed along unmyelinated somatic afferent fibres. Sympathetic activity may release noradrenaline in the peripheral nerve, resulting in changed conductive properties in unmyelinated fibres transmitting sensory information. PMID- 2572145 TI - A double-blind, placebo-controlled study of alpidem, a novel anxiolytic of imidazopyridine structure, in chronically anxious patients. AB - In this double-blind study alpidem, a new imidazopyridine anxiolytic drug, was compared with placebo to assess its efficacy and safety in severely anxious patients at the fixed dose of 150 mg/day (50 mg t.i.d.) for 3 weeks. Fifty-nine patients with a score of at least 18 on the Hamilton Rating Scale for Anxiety (HRSA) entered the trial after a 3- to 7-day placebo run-in period. Symptom improvement was evaluated with the HRSA, the State and Trait Anxiety Inventory (STAI-1 and STAI-2), a Visual Analogue Scale (VAS) and the Clinical Global Impression (CGI). Alpidem was more effective than placebo in improving mean HRSA (total score and factorial scores for somatic and psychic anxiety), STAI-1 and STAI-2 and VAS scores. The efficacy index of the CGI was better for alpidem than for placebo. Side effects were negligible in both groups. Alpidem appears to be a new interesting anxiolytic drug devoid of significant sedative effects on mental functions. PMID- 2572146 TI - A comparison of severe tardive dystonia and severe tardive dyskinesia. AB - The authors present a demographic study comparing tardive dystonia with severe tardive dyskinesia (TD) patients. Tardive dystonia was more common among young men, while severe TD was more common in older women. Neuroleptics were distributed equally in both groups before the onset of the movement disorders. Drug-free periods were common in the history of severe TD than in tardive dystonia patients. PMID- 2572147 TI - Mental and physical capacity and consumption of neuroleptic drugs in residents of homes for aged people. AB - The mental and physical capacity of all residents in homes for aged people were estimated, and their neuroleptic drug consumption and duration of stay were registered. One third was moderately to severely mentally impaired and 38% demanded more extensive nursing care. Physical dependence was significantly associated with mental decline, and less with age. One quarter of severely mentally impaired people had stayed for less than one year; 23% of the residents received neuroleptic drugs. Treatment was more common in physically incapacitated people together with wandering and aggressive ones. Lack of psychogeriatric services may enhance the use of neuroleptic drugs in homes for aged people. PMID- 2572148 TI - Modulation of single cell responses and neuronal interactions produced by iontophoresis of glutamate in adult cat visual cortex. AB - Single neuron responses and interneuronal coordinations after microiontophoretic injections of L-glutamic acid (glutamate) were examined in cats striate cortex with an aid of multielectrode extracellular recording and crosscorrelation analysis. The effects of 15 min long stimulation with glutamate were compared with spontaneous mutability observed within at least 1.5 h. A mutability coefficient was suggested for quantitative data analysis. Spontaneous changes were analyzed in 144 neuronal pairs in two anesthetized and seven pretrigeminal cats. Differences in spontaneous mutability between these groups were not significant. Fluctuations of crosscorrelograms were smaller than fluctuations of poststimulus time histograms (PST). The type of correlogram and receptive field preferred orientation and direction never changed. The effect of glutamate injection was studied in 157 neuronal pairs in four anesthetized and eight pretrigeminal cats. Excitatory responses were observed in 69% of neurons, inhibitory in 18%. The excitatory response to glutamate was accompanied with enhanced visual response in 47% of cases and with reduced visual response in 18%. Crosscorrelogram strength was reduced in 43% of pairs with excitatory response to glutamate and increased only in 23%. After the termination of iontophoresis a persistent increase of the shared input coordination was observed in two pairs in anesthetized animals and in three pairs in pretrigeminal cats. The appearance of direct excitatory coordination was found in one case in pretrigeminal cats. Alterations of PST histograms did not exceed the range of spontaneous mutability. We infer that in a small percent of neuronal pairs the short glutamate stimulation can affect interneuronal links even in adult cat. PMID- 2572149 TI - On a rare atypical form of metachromatic leukodystrophy (MLD): "neurological non mld patients with low levels of arylsulphatase A". Description of two cases. AB - Two young patients (a brother and his sister) with low ASA levels who had myoclonic seizures since the age of eight years, were investigated by means of electrophysiological, neuroradiological and laboratory tests and submitted to a physical examination. In both patients, the disease started at the age of eight years with drug resistent myoclonic seizures. Intelligence was not impaired and ASA levels were lower than normal, but clinical features were not those of classic MLD. Electrophysiological study revealed altered SEPs and normal BAEPs (they are both altered in leukodystrophy). Therefore our findings conclude that tour patients must be considered "Neurological patients non-MLD with low levels of ASA". PMID- 2572151 TI - [Post-traumatic hemarthrosis of the knee joint--an indication for arthroscopy]. AB - Diagnostic arthroscopy including the use of a probe should be performed if the cause for a haemarthrosis of the knee cannot be established by radiological means. The clinical examination of the recently injured knee must be considered to be unreliable due to pain. In approximately one-third of knee injuries an incorrect or incomplete diagnosis is made. Even if ligamentous injuries can be established, arthroscopy is mandatory, because this is the only sure way in which acute and degenerative meniscal or cartilaginous lesions can be diagnosed. Arthroscopy should be performed under general or spinal anaesthesia to allow the use of a tourniquet and the immediate repair of ligaments if deemed necessary. The experienced arthroscopist needs only a few minutes to obtain information that could influence the approach to the injury and its longterm prognosis. In a series of 1238 arthroscopies 252 (20.3%) were done for haemarthrosis of unknown origin. In 68% of these cases diagnosis could only have been established by arthroscopy. In this series of 252 cases 23% had an isolated complete anterior cruciate ligament (ACL) injury, 28% had anterior-medical instability, 8% had partial ACL lesions, 5% posterior cruciate ligament injuries, 15% medial collateral ligament injuries, 11% had only synovial tears or contusions. 5% of cases had chronic ACL instability, 4% had osteochondral fragments which could not be diagnosed radiologically. A recent traumatic patellar dislocation with a tear of the retinaculum was found in 1.6% of the cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572150 TI - [Benign cranial hypertension secondary to treatment with lithium]. AB - A 23 years old woman, diagnosed as having manic-depressive psychosis, in treatment with lithium carbonate for eight months, developed a benign cranial hypertension secondary to the lithium. Suppression of the lithium gave place to the normalization of the pressure of the cerebrospinal liquid which increased again with the reintroduction of the drug. This rare complication of the lithium salts, has only been described in five other cases, all women, with a relative short time of treatment, non-toxic lithium serum levels and without precipitating factors. PMID- 2572152 TI - [Arthroscopy following pediatric injuries of the knee joint--a helpful or superfluous diagnostic procedure?]. AB - Between 1983 and 1987, 1082 arthroscopies were carried out on the knee joint. This included 48 cases of children between the age of seven and fifteen with knee injuries. In 21 cases the clinical diagnosis was confirmed. In 5 cases the clinically indications for an operation could not be confirmed. In 9 cases of arthroscopy, it was demonstrated that an operation was necessary, although this was not clinically diagnosed. Finally in 15 cases, specific conservative therapy could be planned on the basis of the medical evidence. From 35 cases of haemarthrosis, 27 proved to have injuries requiring therapy, 12 of which were not obvious during clinical examination. The results underline advantages of arthroscopy for childhood knee joint injuries, in allowing a definite diagnosis and varying therapeutic measures. PMID- 2572153 TI - [Comparative clinical and arthroscopic study of the knee joint in chondromalacia patellae]. AB - It is not easy to perform clinical diagnosis of chondromalacia patellae. Many cases take a clinically silent course and are discovered by chance only. Analysis for 275 arthroscopies resulted in almost complete agreement between clinical and arthroscopic diagnosis if the damage to the cartilage was diagnosed before arthroscopy. On the other hand, in chondromalacia that remained clinically silent, damage to the cartilage was present in 67% of the cases as detected by arthroscopy. The authors offer detailed comments on the problem of localisation of cartilaginous changes. PMID- 2572154 TI - [Diagnostic value of sonography and double contrast arthrography in meniscus lesions]. AB - Using a real time scanner and a 7.5 mHz transducer, the meniscus can bei visualised as a homogeneous triangle. It is clearly distinguishable from the tibial plane und the condyle of the femur. Tears in the meniscus show up as a double contour rich in echoes with an intervening low-echo area, or as a contour rich in echoes with an adjacent low-echo area. The posterior horn area can be visualised most clearly. Assessment of the interior part of the meniscus is somewhat easier than visualising the exterior part. In a clinical study with 107 menisci examined by sonography and controlled by arthroscopy or arthrotomy the rate of accurate diagnoses by sonography of the meniscus is 82%. 42 of these menisci were additionally examined by double contrast arthrography besides sonography. This yielded an accuracy rate of 74% for double contrast arthrography; the latter is superior to sonography only in the anterior horn area. Problems in respect of meniscus sonography occur only in case of transverse ruptures, scars and longitudinal meniscus ruptures presenting as bucket handle tears near the base. The typical longitudinal tear in the area of the posterior horn can be visualised most clearly. Analysis of the results shows that sonography of the meniscus is a noninvasive, painless and randomly reproducible and risk-free examination method which has a diagnostic value especially in the area of the posterior horn that can be compared with double contrast arthrography. Further studies must show whether with an increasing spread of the method it would be possible to replace double contrast arthrography by sonography in diagnosis of menisceal trouble. PMID- 2572155 TI - [Long-term study of an unusual post-traumatic arthrosis of the ankle joint]. AB - Long-term observation for 15 years of an unusual heavy posttraumatic degenerative arthritis in the upper ankle joint with considerable deformations of the structures and surprising good motility in the joint. Demonstration of the case by pictures and x-ray pictures. The question of overrating posttraumatic degenerative arthritis and its sequels is discussed. PMID- 2572156 TI - [Late results of arthrodeses of the upper ankle joint]. AB - Arthrodesis may be mandatory after destruction or painful arthrosis of the ankle joint. The time at which this should be performed, as well as the technique to be employed, are both judged differently by various writers. We used exclusively the fixateur externe for stabilisation with 44 arthrodeses of the ankle joint, since this ensures stability during exercise and partial load. The complications recorded were two aseptic necroses of the talus and dysostosis in two cases which required corrective surgery. The results obtained by use lead us to the following conclusions: More than one-half of the examined subjects wore the orthopaedic made-to-measure shoes for longer than one year compensating the leg shortening by an average of 1.0 cm. The intraoperatively attempted repositioning of the talus in relation to the axis of the tibia tended to have a rather negative effect in our patients. The at least equal long-term results of early and late arthrodeses indicate that early arthrodeses are preferable, seeing that 55.8% of the followed up persons were able to resume their professional activities relatively quickly. PMID- 2572157 TI - [The value of dynamic adjustment in locking intramedullary nailing]. AB - The so-called "dynamization", i.e. the removal of defined bolts after the first signs of osseous consolidation detected by X-ray film 6 to 10 weeks after surgery employing the static locking nail technique, cannot be translated into reality. In a total of 121 patients in whom static locking nail technique had been employed, dynamization was effected in 23 cases only, i.e. 19%. Complications are likely to occur if the indication for dynamization was wrong or if dynamization is not performed although it was indicated. Dynamization must be prescribed individually on the basis of X-ray film controls and is actually indicated in rare cases only. We refuse to perform it as a routine step after static nail locking of the femur and tibia. PMID- 2572159 TI - [Traumatic diaphragmatic ruptures in an accident surgery patient sample]. AB - The problems of the direct and indirect diaphragm lesions are discussed based on the own collective of patients. The most common trauma pattern was the direct immediate rupture of the left half (Strug). All ruptures were combined with additional trauma including lesions to the abdomen, pelvis, thorax and cranium. Diagnosis could usually be made by conventional X-ray techniques. The high rate of lethality is explained by the extensive additional trauma that is seen with rupture of the diaphragm. PMID- 2572158 TI - [Examples of controlled use of computerized tomography in soft tissue injuries of the area of the head and neck]. AB - CT can be utilised in a selective manner in soft tissue injuries presented in the head, neck and shoulder regions. PMID- 2572160 TI - [Metal implants of the coxal femur in the elderly patient should not be removed]. PMID- 2572161 TI - Effect of epidermal growth factor on non-steroidal anti-inflammatory drug-induced intestinal damage. AB - Epidermal growth factor (EGF, 10 micrograms/kg po, ip, or sc, BID, and 20 micrograms/kg iv) had no protective activity in the indomethacin-induced intestinal lesion model (6 h model). In the ethanol-induced gastric lesion model, EGF (10 micrograms/kg sc) reduced lesions by 52% and reduced gastric acid secretion by 68% (5 micrograms/kg iv). In the 24 h indomethacin-induced intestinal lesion model, pretreatment with EGF (10 micrograms/kg sc, BID; 1 day before and during indomethacin treatment) had no beneficial effects. Therefore, EGF had no protective effects against non-steroidal antiinflammatory drug (NSAID) induced intestinal lesions at doses that protect against the necrotizing action of ethanol and that inhibit gastric acid secretion in the rat. PMID- 2572162 TI - [Effects of combined instillation of dapiprazole and epinephrine on human anterior chamber depth, intraocular pressure and pupil diameter]. AB - Pilocarpine, a first choice drug for primary angle closure glaucoma (PACG), is reported to produce shallow anterior chamber and posterior synechia and consequently a small immobile pupil. In this paper, dapiprazole, a newly synthesized alpha 1 blocker, was given topically in combination with epinephrine eye drops to 15 healthy volunteers and 10 patients with PACG. In both volunteers and patients, the treated eyes revealed deeper anterior chambers, thinner lenses and significant IOP reduction after the combined instillation of dapiprazole and epinephrine. Though light reaction was preserved, mild miosis was observed in volunteers, but no pupillary change in patients. The conjunctival hyperemia induced by dapiprazole was reversed by additional instillation of epinephrine. Burning sensation was the only side effect observed after instillation of dapiprazole. In conclusion, the combined instillation of dapiprazole and epinephrine appears to be a good choice for the therapy of PACG. PMID- 2572163 TI - [A case of persistent mullerian duct syndrome associated with seminoma]. AB - A case of persistent Mullerian duct syndrome associated with seminoma is presented. A 28-year-old man was hospitalized with the complaint of a palpable mass in the left lower abdomen and lumbago. Physical examination revealed a normal right scrotal testicle and a small right inguinal hernia that did not contain a mass, but no testicle was found in the left scrotum. Laparotomy was performed for suspected tumor of an intraabdominal testis. At abdominal exploration the left testis was found to be cross-ectopic to just behind the pubic symphysis and to be replaced completely with a large tumor, which was supplied by the left spermatic vessels and had a vas deferens emptied to the right seminal vesicle. In addition another cord-like structure was traced into the right inguinal canal. The tumor was dissected with these structures. On histological examination the tumor was pure seminoma and the cord-like structure proved to be an immature uterus, Fallopian tubes and seminal vesicle. PMID- 2572164 TI - Takayasu arteritis: angiographic findings and results of angioplasty. AB - Takayasu arteritis is a systemic disease characterized by occlusion of the aorta and its branches. We performed coronary angiography and thoracic and abdominal aortography on 47 patients with Takayasu arteritis. Angiographic findings included arterial stenosis, occlusion, or aneurysm. The left subclavian artery was involved in 26 cases (55%), the abdominal aorta in 25 cases (53%), the right renal artery in 21 cases (45%), the right subclavian and left renal arteries in 18 cases (38%), the descending thoracic aorta in 15 cases (32%), and the left common carotid artery in 14 cases (30%). The coronary arteriograms in the 47 patients showed coronary involvement in seven (15%). Proximal or osteal lesions were present in six cases. Percutaneous transluminal angioplasty was performed successfully in eight patients for treatment of three aortic lesions, nine renal artery lesions, one subclavian artery lesion, and one coronary artery lesion. In the three cases with recurrence after 4-7 months, repeated angioplasty was successful. For appropriate management of Takayasu arteritis, we suggest thorough angiographic evaluation and proper intervention, including percutaneous transluminal angioplasty in selected cases for revascularization. PMID- 2572165 TI - Improvement of diastolic function after reversal of left ventricular hypertrophy induced by long-term antihypertensive treatment with tertatolol. AB - In 15 previously untreated hypertensive subjects with left ventricular (LV) hypertrophy who responded favorably (supine blood pressure less than or equal to 140/90 mm Hg) to antihypertensive treatment with a nonselective beta-blocking agent, tertatolol, the effects of reversal of LV hypertrophy on systolic and diastolic function were assessed. Patients underwent echocardiographic and radionuclide studies in control conditions (phase 1), after 1 month of blood pressure normalization (phase 2), after reversal of LV hypertrophy or at least a 20% reduction of LV mass compared to basal value (phase 3) and finally, after a 1 month washout (phase 4). In phase 2, blood pressure (130 +/- 2/85 +/- 1 vs 148 +/ 4/104 +/- 1 mm Hg) and heart rate (59 +/- 1 vs 76 +/- 2 beats/min) decreased (both p less than 0.01); LV mass remained unchanged. There were improvements in peak filling rate (end-diastolic volume/s) (2.4 +/- 0.1 vs 2.0 +/- 0.1), ejection fraction (65 +/- 1 vs 61 +/- 1%) and their ratio (stroke counts/s) (3.7 +/- 0.2 vs 3.2 +/- 0.1) (all p less than 0.05). In phase 3, blood pressure and heart rate were unchanged and reversal of LV hypertrophy was accompanied by a further increase in peak filling rate (2.9 +/- 0.1), ejection fraction (69 +/- 1%) and their ratio (4.1 +/- 0.1) compared to phase 2 (all p less than 0.01). Finally, in phase 4 blood pressure and heart rate returned to the basal value, but peak filling rate (2.7 +/- 0.1) and ejection fraction (65 +/- 1%), although reduced compared to phase 3, were still higher than phase 1.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572166 TI - Ammonia transport in the proximal tubule in vivo. AB - Studies were performed to characterize the determinants of proximal tubule ammonia entry (and retention) in vivo. Rat proximal tubules were studied in vivo using in situ microperfusion. In both normal animals and animals with metabolic acidosis, increasing luminal flow rate significantly enhanced luminal ammonia entry. In contrast, luminal pH was not as important in determining ammonia entry. Analysis of the levels of luminal NH3 in these studies was not consistent with simple diffusion equilibrium of NH3. In animals with chronic metabolic acidosis, additional studies demonstrated that inhibition of the Na+-H+ exchanger had no direct effect on luminal ammonia entry. However, studies of ammonia efflux from tubules perfused with 10 mmol/L ammonia demonstrated significant transport of both NH3 and NH4+. Studies of luminal glutamine deamidation via gamma glutamyltransferase in control conditions did not indicate a significant role for luminal ammoniagenesis in the superficial proximal tubule in vivo. These and other recent studies of proximal tubule ammonia transport significantly modify the traditional diffusion equilibrium (of NH3) model of ammonia transport. Luminal flow rate is an important determinant of luminal ammonia entry. Transport of NH4+, both into and out of the tubule lumen, represents a major component of total ammonia transport. PMID- 2572167 TI - Experience with a two-tiered therapeutic interchange policy. AB - A university hospital's formulary policy for therapeutic interchange is described in which pharmacists can routinely interchange some drugs but must contact the prescribers before interchanging other drugs. For drugs that are not automatically interchanged by the pharmacy, the formulary contains a "class representative," which pharmacy may change as relative prices of drug products change. When a non-formulary drug for which there is a designated class representative is prescribed, pharmacy contacts the prescriber. When a class representative for injectable histamine H2-receptor antagonists was being selected, previous positive and negative experiences with establishing therapeutic equivalence for antimicrobial agents were considered. The implementation of H2 antagonist therapeutic equivalence included the following steps: determining potential cost savings, reviewing the literature, consulting with specialty practitioners, presenting the information to the pharmacy and therapeutics committee, distributing formal bids, and educating hospital staff. Before cimetidine was designated the class representative, 84% of orders for injectable H2 antagonists were for ranitidine; one year later, 90% were for cimetidine. Orders for oral H2 antagonists also changed from predominantly ranitidine to predominantly cimetidine. The hospital's total costs for H2 antagonists decreased 8.4% in one year. The two-tiered approach to therapeutic interchange can reduce drug costs and increase the scope of agents deemed therapeutic equivalents in a manner that is acceptable to physicians and pharmacists. PMID- 2572168 TI - Molecular genetics of inherited antithrombin III deficiencies. AB - The cloning of antithrombin III (ATIII) complementary deoxyribonucleic acids and the determination of the ATIII gene structure have permitted a systematic evaluation of the molecular basis for inherited ATIII deficiencies. Sixteen kindreds with the most common form of the deficiency, in which plasma ATIII antigen levels and activity are proportionately reduced, were studied. Two polymorphic deoxyribonucleic acid markers were used to resolve parental ATIII alleles and to trace their inheritance patterns. In 15 of 16 cases, the structure of the affected ATIII allele was indistinguishable from normal, suggesting that relatively small mutations, resulting in gene inactivation, are responsible for the lower ATIII levels in these affected families. In the remaining kindred, complete deletion of one ATIII allele was seen. Also investigated was the molecular basis for a qualitative form of ATIII deficiency in a French-Canadian family with normal levels of immunoreactive protein but only half the expected levels of serine protease inhibitor activity. Using polymorphic markers, the abnormal allele was identified, cloned, and partially sequenced from the propositus. A single G----A transition was seen in the first base of codon 382, resulting in an alanine----threonine substitution in the defective protein. This mutation, together with others in this vicinity, defines a minimal length for a fully functional thrombin-binding domain. PMID- 2572170 TI - Spontaneous rupture of a renal artery aneurysm in polyarteritis nodosa: critical review of the literature and report of a case. PMID- 2572169 TI - Detection of X-linked lymphoproliferative disease using molecular and immunovirologic markers. AB - PURPOSE, PATIENTS, AND METHODS: Detection of males affected with the X-linked lymphoproliferative disease (XLP) was sought using immunovirologic and molecular genetic linkage techniques. The study population consisted of 20 males in six families with XLP. RESULTS: Concordance for detection of affected males was 100% when linkage analysis using DXS42 and DXS37 DNA probes and antibody responses to challenge with bacteriophage phi X174 were both determined. Most affected males showing IgG subclass immune deficiency could not produce antibodies to Epstein Barr virus nuclear antigen and had deficient responses to challenge with bacteriophage phi X174. CONCLUSION: Use of only one of the techniques described can fail to lead to the diagnosis of XLP, because problems can prevail with each individual determination. PMID- 2572171 TI - Takayasu's arteritis as a cause of fever of unknown origin. PMID- 2572172 TI - Cholinergic control of smooth muscle peristalsis in the cat esophagus. AB - The aim of this study was to examine in detail the effects of selective cholinergic and other pharmacological antagonists on primary and secondary peristalsis in the smooth muscle of the cat esophagus in order to fully characterize the cholinergic contribution to peristalsis in this species. Primary and secondary peristalsis in the smooth muscle part of the feline esophagus was completely abolished by atropine, 4-dephenylacetoxy-N-methylpiperidine methiodide (4-DAMP) (a selective M2 muscarinic antagonist), hexamethonium, and high doses of nicotine. Pirenzepine (a selective M1 muscarinic antagonist), propranolol, and phentolamine were without effect, as were naloxone, methysergide, and pyrilamine. From these findings we conclude that primary and secondary peristalsis in feline esophageal smooth muscle involves nicotinic ganglionic neurotransmission as well as postganglionic release of acetylcholine that acts directly on muscarinic receptors located on the smooth muscle. Peristalsis in esophageal striated muscle does not involve either synaptic transmission or muscarinic receptors. PMID- 2572173 TI - Putative mechanisms involved in excitatory and inhibitory effects of somatostatin on intestinal motility. AB - The mechanism of action of somatostatin on the motility of intestine was examined in the entire preparation and the longitudinal muscle attached with Auerbach's plexus (LA) preparation of guinea pig ileum, in relation to the cholinergic neuron and gamma-aminobutyric acid (GABA)ergic neuron. Somatostatin produced a transient potentiation of electrical stimulation-induced twitch contractions followed by an inhibition. The excitatory effect of somatostatin was associated with an increase in the release of [3H]acetylcholine (ACh) from the preparations preloaded with [3H]choline. Bicuculline, a GABAA antagonist, inhibited the somatostatin-induced excitatory effect. Somatostatin inhibited the electrical stimulation-induced twitch contraction and release of [3H]ACh, and the inhibition was greater in the entire preparation than in the LA. Phaclofen, a GABAB antagonist, prevented the inhibitory effects of somatostatin. Somatostatin induced a Ca2+ -dependent, tetrodotoxin-sensitive release of [3H]GABA from the preparations preloaded with [3H]GABA. Therefore somatostatin exerts excitatory and inhibitory effects on the cholinergic neuron due to the stimulation of the GABAergic neuron, and the motility of the intestine is regulated. PMID- 2572174 TI - Regulation of cellular glutathione. AB - In addition to its participation in a variety of other biochemical reactions, glutathione (GSH) is a major antioxidant. It is regularly generated intracellularly from its oxidized form by glutathione reductase activity that is coupled with a series of interrelated reactions. Synthesis of GSH also takes place intracellularly by a two-step reaction, the first of which is catalyzed by rate-limiting gamma-glutamylcysteine synthetase activity. Intracellular substrates for GSH are provided both by direct amino acid transport and by a gamma-glutamyl transpeptidase reaction that salvages circulating GSH by coupling the gamma-glutamyl moiety to a suitable amino acid acceptor for transport into the cell. Although the liver is a net synthesizer of circulating GSH, organs such as the kidney salvage GSH through the gamma-glutamyl transpeptidase reaction. Intracellular GSH may be consumed by GSH transferase reactions that conjugate GSH with certain xenobiotics. Elevation of cellular GSH levels in cultured cells in response to hyperoxia or electrophilic agents such as diethylmaleate is coupled with an increase in activity of the Xc- transport system for the amino acids cystine and glutamate. Strategies may be developed for protection against oxidant injury by enhancement of transport systems for precursor amino acids of GSH or by providing substrate that circumvents feedback inhibition of GSH synthesis. PMID- 2572175 TI - Effects of alpha-adrenergic agonists on intracellular and intramitochondrial pH in rat proximal nephrons. AB - Rat proximal tubular segments were used to examine alpha-adrenoceptor alterations in Na+-H+ exchange by monitoring intracellular pH (pHi) and mitochondrial matrix pH (pHm). To obtain pHi, tubules were incubated with the cell-permeant fluorescent probe, 2',7'-bis(2-carboxyethyl)-5(6) carboxyfluorescein acetoxymethyl ester in a HCO3--free Na+ buffer. The intracellular distribution of the weak acid [2-14C] 5,5-dimethyloxazolidine-2,4-dione was used to calculate pHm, using values of medium pH, pHi, cell volume, and matrix content. Several selective alpha 1- and alpha 2-adrenoceptor agonists and the endogenous mixed agonist, norepinephrine, all produced dose-related increases in pHi. With each of the agonists tested, a maximum increase in pHi was observed at 1 microM final concentrations, with peak effects occurring in less than 1 min. Pretreatment with ethylisopropyl amiloride (EIPA, 10 microM), a specific inhibitor of proximal Na+ H+ exchange, blocked receptor-stimulated increases in pHi, as well as stimulation of Na+-H+ exchange by phorbol ester (PMA, 0.1 microM). Similarly, selective alpha 1- (prazosin, 0.1 microM) and alpha 2-(idazoxan, 0.1 microM) adrenoceptor antagonists inhibited alterations in agonist-induced pHi changes, whereas PMA stimulated increases in pHi remained unaffected. Neither alpha 1- nor alpha 2 adrenoceptor agonists produced differences in pHm. Adrenoceptor agonist-induced pHi changes were also assessed at various concentrations of external Na+ (0-135 mM). It was observed that 0 and 10 mM external Na+ concentrations significantly reduced alpha 1- and alpha 2-adrenoceptor-stimulated pHi changes; Km values for the alpha 1-agonist phenylephrine and the alpha 2-agonist B-HT 933 were 18.0 +/- 2.1 and 22.7 +/- 2.6, respectively. In summary, stimulation by alpha-adrenergic agonists may be blocked at the receptor level with specific alpha-antagonists or at the exchanger with EIPA. The increase in cellular pH induced by these agonists is sensitive to external Na+ and reflects alpha-adrenoceptor activation of the Na+-H+ exchanger. PMID- 2572176 TI - Dilator effects of amino acid neurotransmitters on piglet pial arterioles. AB - We examined the effects of topically applied amino acids (glutamate, aspartate, glycine, and taurine) and a synthetic glutamate analogue [N-methyl-D-aspartate (NMDA)] on pial arteriolar tone and cortical surface cerebrospinal fluid (CSF) dilator prostanoid concentrations in anesthetized newborn pigs. We also determined whether prolonged contact of pial arterioles with glutamate (10(-3) M) and aspartate (10(-3) M) would alter arteriolar responses to exogenous isoproterenol or norepinephrine. Vascular responses were determined using the closed cranial window technique and intravital microscopy. Concentrations of prostaglandin E2 and 6-ketoprostaglandin F1 alpha in CSF under the cranial window were determined using radioimmunoassay. At the highest dose tested (10(-3) M), NMDA dilated arterioles by 30 +/- 4% (n = 8), glutamate by 21 +/- 5% (n = 6), aspartate by 28 +/- 10% (n = 5), and taurine by 21 +/- 2% (n = 7). Glycine application did not change pial arteriolar diameter significantly (n = 8). The amino acids tested (NMDA and glutamate) did not increase CSF levels of dilator prostagnoids, and intravenous indomethacin trihydrate did not change vascular responsiveness to NMDA. Furthermore, dilator responsiveness to isoproterenol and constrictor responsiveness to norepinephrine were not affected significantly after 30 min of topical application of glutamate and aspartate to the pial surface (n = 4). We conclude that these amino acids are potent dilators of the neonatal cerebral circulation. The mechanism of dilation in the cases of NMDA and glutamate does not appear to involve dilator prostanoids. Furthermore, prolonged contact with excitatory amino acids under these conditions does not alter subsequent cerebrovascular responsiveness. PMID- 2572177 TI - Central interleukin 1-elicited hyperinsulinemia is mediated by prostaglandins but not autonomics. AB - This laboratory previously reported that centrally administered interleukin 1 (IL 1) in fasted pentobarbital-anesthetized rats elicited significant hyperinsulinemic and febrile responses. In characterizing this putative central mechanism for the regulation of pancreatic insulin secretion, hyperinsulinemia and fever elicited by IL-1 injected intravenously (iv) or intracerebroventricularly (icv) was totally eliminated by prior cyclooxygenase inhibition with indomethacin, ibuprofen, or meclofenamate but not lipoxygenase inhibition with propyl gallate or leukotriene receptor antagonism with LY 171883. Furthermore, central administration of prostaglandin E2 at 10 and 100 ng doses consistently evoked hyperinsulinemic, hypercorticotropinemic, and febrile responses in anesthetized rats maintained on isothermal pads. beta-Adrenergic and vagus nerves to the pancreatic beta-cells seemed likely candidates to mediate the enhanced secretion of insulin elicited by IL-1 acting centrally. However, pretreatment of rats with hexamethonium, propanolol, atropine, or bilateral subdiaphragmatic vagotomy all failed to reduce hyperinsulinemia after IL-1 iv or icv. This evidence suggests that the central mechanism for enhanced pancreatic insulin secretion elicited by IL-1 may depend on a humoral rather than autonomic neural efferent pathway. Moreover, the hyperinsulinemia is mediated in part by prostaglandins just like the well-studied febrile response. PMID- 2572179 TI - Genetic heterogeneity in Peruvian Leishmania isolates. AB - Leishmania were isolated from Peruvian patients with uta or espundia; genomic DNA was examined for restriction fragment length differences by Southern blot analysis using DNA probes for beta-tubulin and for the major surface antigen gp63. Using 5 different restriction endonucleases, Peruvian isolates show homogeneity when examined at the beta-tubulin locus. In contrast, the organisms demonstrated heterogeneity both within and between disease groups when examined for restriction site differences within the gp63 locus. The differences observed did not correlate with the 2 disease groups. Comparison of these Peruvian isolates to New World reference strains of the Leishmania braziliensis complex reveals no consistent pattern of identity with either L. b. guyanensis, L. b. panamensis, or L. b. braziliensis. PMID- 2572178 TI - Ecologic studies of mosquitoes and birds as hosts of Ockelbo virus in Sweden and isolation of Inkoo and Batai viruses from mosquitoes. AB - Field studies were conducted in central Sweden from 1983 through 1985 to obtain information on the etiologic agent of Ockelbo disease, described in Sweden in the 1960s and probably identical to Pogosta disease in Finland and to Karelian fever in the western USSR. Mosquitoes (63,644) collected during this 3 year period yielded 21 virus strains. Ockelbo virus isolations were from Culiseta morsitans (5 strains), Culex pipiens and/or Cx. torrentium (6 strains), and Aedes cinereus (3 strains). Inkoo (6 strains) and Batai (1 strain) viruses were recovered from Ae. communis. Blood samples collected March-May from migrating birds on the southeast and est coast of Sweden and in July and August from resident birds in east-central Sweden were tested for neutralizing antibody to Ockelbo virus. Antibody was not detected in 328 birds sampled during spring migrations. Two of 58 (3.4%) birds bled in July and 8 of 78 birds (10%) bled in August had antibody to Ockelbo virus. Ockelbo virus circulates in a mosquito-bird-mosquito cycle, with Cs. morsitans and Cx. pipiens and/or Cx. torrentium as enzootic vectors. Antibody was detected in passerine birds. Other classes of birds or other vertebrates were not sampled. Aedes cinereus may serve primarily to transmit virus to people. The role of other mosquito species as vectors for people is unknown. PMID- 2572180 TI - Assessment of the transferrin index in screening heavy drinkers from a general practice. AB - The aim of this study was to assess the performance of the transferrin (Tf) index in screening heavy drinkers and to compare its performance to that of gamma glutamyl transpeptidase (GGT). Tf index, GGT, and transaminase activities were determined in a group of 173 subjects (49% males) recruited in a family doctor's practice (age: 54 +/- 1.5 yr; daily alcohol intake: 32 +/- 3.5 g; mean +/- SEM). Tf subfractions were quantified by isoelectric focusing and immunofixation on polyacrylamide gel. The Tf pl 5.7 to 5.4 ratio, or Tf index, was used as a marker of excessive drinking, with a cut-off point at 7%. Alcohol consumption was assessed through a face-to-face interview. Excessive drinkers were defined as those with a daily alcohol intake greater than 80 g over at least 2 years; 20 were classified as excessive drinkers (alcohol consumption: 92-232 g/day). All but four had normal transaminase activities indicating the low prevalence of hepatic impairments in this sample. The Tf index was found to have a sensitivity of 45%, specificity 89%, positive predictive value 35%, negative predictive value 92%. The corresponding results for GGT were 52%, 80%, 27%, and 92%, respectively. Concordance between the Tf index and GGT was assessed by the kappa coefficient (kappa) which was 0.22 indicating poor agreement between the two markers in selecting excessive drinkers (perfect association: kappa = 1, no association: kappa = 0). PMID- 2572181 TI - Topical levocabastine versus sodium cromoglycate in allergic conjunctivitis. AB - The aim of this study was to evaluate the effect of levocabastine, a new H1 blocking antihistamine for topical use, in comparison with sodium cromoglycate on conjunctival symptoms of birch pollinosis. The two drugs were compared in a randomized double-blind comparative study over 5 weeks in 37 children and adolescents (6-19 years of age) with birch pollen conjunctivitis. Nasal symptoms occurred in 31 of the children and were treated with beclomethasone dipropionate nasal spray. An oral antihistamine was offered as rescue medication for eye symptoms. Initially, the patients received placebo four times a day for a 7-day run-in period. Conjunctival symptoms were recorded daily on diary cards on a 100 mm visual analogue scale. The pollen counts indicated a short but intensive birch pollen season. There was no statistically significant difference between the two treatment groups with regard to eye symptom scores before and during active treatment. However, the patients' evaluation of the efficacy of the therapy was in favour of levocabastine (P less than 0.01). Topical levocabastine, an H1 blocker, applied twice daily, seems to protect from symptoms of allergic conjunctivities as favourably as sodium cromoglycate applied four times a day. There was no difference in number or character of reported adverse reactions between the two treatment groups. PMID- 2572182 TI - Efficacy and safety of loratadine suspension in the treatment of children with allergic rhinitis. AB - The safety and efficacy of loratadine was compared with that of dexchlorpheniramine in children with allergic rhinitis. Twenty-one children received loratadine 0.11-0.24 mg/kg ideal body weight once daily and 19 dexchlorpheniramine 0.10-0.23 mg/kg every 8 h (0.30-0.69 mg/24 h) for 14 consecutive days. Both loratadine and dexchlorpheniramine were effective in reducing nasal and ocular symptoms in allergic children. Substantial improvement in allergy symptoms was observed at the first evaluation (day 3 of treatment) and was maintained for the study duration. No significant trend of abnormality in laboratory parameters was observed. Drowsiness was present only in the dexchlorpheniramine-treated group. Loratadine appears to be a simple, effective and safe therapy for seasonal allergic rhinitis. PMID- 2572184 TI - Effect of rate of injection on the neuromuscular block produced by vecuronium. AB - A subparalytic dose (0.015 mg/kg) of vecuronium bromide was administered to matched pairs of patients undergoing routine dental surgery under enflurane nitrous oxide/oxygen anesthesia either as a rapid bolus injection or as a slow infusion over 5 min. It was demonstrated that bolus injection produced peak plasma levels of drug considerably greater than those following slow infusion. Time to maximum block was more rapid following bolus (368 +/- 84 [SD] sec) than by infusion (615 +/- 88 sec), but the maximum block produced, either demonstrated by the reduction in amplitude of T1 from control or the T1:T4 ratio on the integrated electromyogram (Datex IEMG), was similar in each group irrespective of rate of injection, T1 and T4 referring to the first and last twitch in a train of four series. PMID- 2572185 TI - Esmolol in the management of epinephrine- and cocaine-induced cardiovascular toxicity. PMID- 2572183 TI - Placental toxicity in rats after administration of synthetic glucocorticoids. A morphological, histochemical and immunohistochemical investigation. AB - Administration of the synthetic glucocorticoids dexamethasone and triamcinolone to pregnant rats between gestational day (GD) 16 and 20 caused dose-dependent placental lesions on GD 21 and 22 which were detected by morphological, histochemical and immunohistochemical means. Maternal blood spaces, trophoblast layer and fetal blood vessels were altered primarily in the centre of the placental labyrinth. Less severe changes were found in the junctional zone, chorionic plate and intraplacental yolk sac. On GD 21, low doses increased the amount of glycogen, while high doses induced a loss of glycogen. gamma-glutamyl transpeptidase activity was increased in the spongiotrophoblast and the labyrinthic trophoblast and dipeptidyl peptidase IV activity in fetal capillary endothelium, whereas alpha-glutamyl aminopeptidase and microsomal alanyl amino peptidase were not affected. Additionally, in the fetal capillary endothelium an increase of immunoreactivity for the von Willebrand factor occurred. These data suggest that synthetic glucocorticoids affect placental tissues at different and rather specific levels, which may in turn disturb placental function and contribute to fetal maldevelopment. PMID- 2572187 TI - Loratadine, a non-sedating H1-receptor antagonist (antihistamine) PMID- 2572186 TI - Comparison of the effects of oral famotidine and ranitidine on gastric volume and pH. PMID- 2572188 TI - Treatment of the recalcitrant asthmatic. AB - Treatment of "difficult" asthma requires good doctor-patient communication, patient education, and attention to precipitating factors as well as an aggressive drug regimen. Specific medications include inhaled sympathomimetic and anti-cholinergic bronchodilators, inhaled, oral and/or intravenous corticosteroids and, in certain circumstances, mast cell stabilizing drugs such as cromolyn sodium. The use of systemic theophyllines is currently undergoing critical reevaluation. There have been a number of recent developments in the search for steroid-sparing agents and drugs that inhibit inflammatory mediators felt to be important in the pathophysiology of asthma. Most of these drugs are still undergoing evaluation in multicentre clinical trials. The newer antiinflammatory agents, methotrexate and gold, should probably not be used on a routine basis except as part of randomized, ethically approved clinical trials. PMID- 2572189 TI - Comparison of efficacy and tolerability of terfenadine administered once daily versus twice daily in patients with chronic idiopathic urticaria. AB - The objective of this double-blind study, which is part of a multicenter study, was to determine whether terfenadine (120 mg once daily) has similar efficacy and tolerability to the standard dosage of 60 mg twice daily in the treatment of chronic urticaria. Forty-one patients were randomly allocated to two parallel groups and treated for 2 weeks with either regimen. Evaluation of efficacy was based on rating scales for investigator and patient. Primary endpoints were itch, number of wheals, wheal size, and the overall rating of efficacy. A similar improvement was seen in all variables. There were no statistically significant differences between groups. Both treatments were well tolerated. PMID- 2572190 TI - Infectious bovine keratoconjunctivitis epizootic associated with area-wide emergence of a new Moraxella bovis pilus type. AB - Pilus-mediated adherence is a virulence attribute of Moraxella bovis. Several pilus types have been shown to exist among strains of this bacterium, but correlation between pilus type and specific field cases of the disease has not been done. During the summer of 1987, an epizootic of infectious bovine keratoconjunctivitis was reported in 7 Iowa counties. Eight isolates of M bovis were secured from 12 episodes studied. All 8 of the isolates were nearly homogeneous in biochemical properties and had the same plasmid biotype. Pilus typing performed by immunofluorescence and immunogold electron microscopy identified a single new pilus type among 5 of the 8 isolates. This pilus type was identified in field cases that developed within a narrow time frame and over large distances. The implication of these findings is that infectious bovine keratoconjunctivitis epizootics may be associated with emergence of a novel pilus type, and that rapid dissemination over wide distances can occur, presumably by transportation of carrier cattle. PMID- 2572191 TI - Effects of idazoxan, tolazoline, and yohimbine on xylazine-induced respiratory changes and central nervous system depression in ewes. AB - We compared the ability of 3 alpha 2-adrenoreceptor antagonists, idazoxan (0.05 mg/kg), tolazoline (2 mg/kg), and yohimbine (0.2 mg/kg) to reverse xylazine (0.3 mg/kg)-induced respiratory changes and CNS depression in 6 ewes. Once weekly, each ewe was given a random IV treatment of xylazine, followed in 5 minutes by either an antagonist or 0.9% NaCl solution. Xylazine alone caused recumbency for 54.2 +/- 5.3 minutes (mean +/- SEM). Xylazine also increased respiratory rate and decreased PaCO2 for at least 45 minutes, but did not significantly change arterial pH or PaCO2. Idazoxan and tolazoline were equally effective in reversing the respiratory actions of xylazine; however, yohimbine was less effective in reducing the respiratory rate and was ineffective in antagonizing the decreased PaO2. Idazoxan and tolazoline decreased the duration of xylazine-induced recumbency to 6.3 +/- 0.6 and 9.5 +/- 2.3 minutes, respectively, whereas yohimbine did not significantly change this effect of xylazine. Thus, at the dosages studied, idazoxan and tolazoline appeared to be more effective than yohimbine in reversing the respiratory and CNS depressant actions of xylazine in sheep. PMID- 2572192 TI - The effect of an increase in inhaled allergen dose after rimiterol hydrobromide on the occurrence and magnitude of the late asthmatic response and the associated change in nonspecific bronchial responsiveness. AB - We have used a short-acting beta 2-adrenoceptor agonist, rimiterol hydrobromide, to allow a larger dose of allergen to be administered to previous single "early allergen responders" to investigate if an increased dose of allergen could induce a late asthmatic response (LAR) and whether this would influence the subsequent level of allergen-acquired nonspecific bronchial hyperresponsiveness. Pretreatment with inhaled rimiterol hydrobromide 400 microgram enabled an increase in allergen dose inhaled by a geometric mean of 8.9-fold (range, 2 to 29.1) in eight atopic subjects with mild asthma who initially were classified as single early responders with a maximal fall in FEV 3 to 8 h after allergen challenge (Lmax) of less than 15% from baseline value. The magnitude of the early asthmatic response was similar to that obtained on the control day when allergen challenge was performed in the absence of rimiterol hydrobromide. Five subjects were converted to dual allergen responders with Lmax of greater than 15% from premedication baseline value. For the whole group, there was a significant increase in the magnitude of LAR whether calculated as Lmax (p less than 0.05) or as area under the FEV1 time response curve between 3 and 8 h postchallenge (p less than 0.01). The provocation concentrations of methacholine causing a 20% fall in FEV1 (PC20) at 8 h postchallenge were significantly reduced on both days when compared with the corresponding prechallenge values (p less than 0.05 on control day, p less than 0.01 on rimiterol day). However, despite the increase in the magnitude of LAR on the rimiterol day, the reduction in postchallenge PC20 did not differ significantly from that occurring on the control day.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572193 TI - [Dyslipoproteinemias and coronary disease. Genetic markers]. AB - Atherosclerosis and its consequences are a significant cause of death in industrialized countries. In recent years, due to modern techniques which permit DNA analysis, a series of alleles associated with dyslipoproteinemia and heart disease have been identified. In this study these genetic markers, in particular those found at the level of the genes of apolipoproteins, are reviewed. The knowledge of these facts is important for the paediatrician, since these disturbances can be identified in the first days of an individual's life, thus a positive influence can be made over the lifestyle habits. PMID- 2572194 TI - Secretin and calcium provocative tests in the Zollinger-Ellison syndrome. A prospective study. AB - STUDY OBJECTIVE: To evaluate criteria of positivity for and usefulness of both the secretin and calcium gastrin-provocative tests in patients with the Zollinger Ellison syndrome. DESIGN: Prospective trial in consecutive patients. SETTING: Referrals to a clinical research center. PATIENTS: Consecutive sample of 80 patients with the Zollinger-Ellison syndrome. INTERVENTION: Kabi-secretin (2 U/kg body weight) given by intravenous bolus and calcium gluconate (10%) (54 mg/kg.h [5 mg/kg.h of calcium]) given by continuous intravenous infusion for 3 hours. Serum gastrin measured at -15, and -1 minutes before, and 2, 5, 10, 15, 20, and 30 minutes after secretin, or every 30 minutes for 3 hours during the calcium infusion. Serum calcium and serum gastrin were measured simultaneously during the calcium infusion. MEASUREMENTS AND MAIN RESULTS: There was no significant difference in the responses of patients with different extents or locations of the tumor, presence or absence of multiple endocrine neoplasia, type-I, or with fasting gastrin less than or greater than 1000 pg/mL. In patients with fasting gastrin of less than 1000 pg/mL, the sensitivity of the secretin test using the criterion of an increase in gastrin of at least 110 pg/mL was 93% (CI, 76% to 99%) and for an increase of 200 pg/mL it was 85% (CI, 66% to 96%), (P greater than 0.05). With the calcium infusion test, the sensitivity using the criterion of an increase of 395 pg/mL was 43%, (CI, 23% to 66%) and for an increase of 50% was 74% (CI, 52% to 90%), (P less than 0.01). The calcium infusion test was positive in 33% of patients with a negative secretin test. With the secretin test, 75% of patients had a positive response by 5 minutes, 95% by 10 minutes, 100% by 15 minutes, and 6% only at 2 minutes. With calcium infusion, patients had positive responses at 120 to 180 minutes. CONCLUSIONS: The secretin test is preferred over the calcium test because of its greater sensitivity and simplicity. The recommended criteria are a 200 pg/mL increase for the secretin test and a 395 pg/mL increase for the calcium test. The calcium test should be reserved for patients having a negative secretin test, gastric acid hypersecretion, and a strong clinical suspicion of the Zollinger-Ellison syndrome. PMID- 2572195 TI - Organization and expression of homeobox genes in mouse and man. PMID- 2572196 TI - Somatostatin treatment of gastrointestinal fistulae. AB - Output from gastrointestinal fistulae to the skin was measured in five patients during total parenteral nutrition and restricted oral intake. Supplemental infusion of somatostatin during six days caused a significant reduction in discharge in all patients compared to the pretreatment level, but none of the fistulae closed as a result of the treatment. Somatostatin may be considered an adjuvant drug to parenteral nutrition and restricted oral intake in the conservative treatment of external fistulae from the gastrointestinal tract. PMID- 2572197 TI - Scrapie prions. PMID- 2572198 TI - Preliminary evidence for inhibitory effect of glycyrrhizin on HIV replication in patients with AIDS. AB - Glycyrrhizin (GL) at a dose of 400-1600 mg/day (7.2-30.8 mg/kg/day) was administered intravenously for a period of more than a month, on 6 separate occasions, to 3 hemophiliacs with acquired immune deficiency syndrome (AIDS). Human immunodeficiency virus type 1 (HIV-1) p24 antigen was detected at the beginning of 5 of the 6 treatment courses. Viral antigen was not detected at the end of or during 3 of the 5 treatment courses and decreased to a low level following the 2 other courses. These findings suggest that GL might inhibit HIV-1 replication in vivo. PMID- 2572199 TI - [Glycoproteins associated with metastatic potential of cancer]. AB - Carbohydrate moieties of glycoproteins have been implicated to be involved in cellular adhesion. Therefore, certain carbohydrate structures in glycoproteins are expected to be associated with metastatic behaviour of cancer cells; such carbohydrate structure can be used as an indicator for the treatment of cancer patients based on the predicted metastatic potential of the cancer cells. The following results recently reported is interesting from the above view point. In certain cancer cells of the mouse, beta 1----6 branching of asparagine-linked oligosaccharides, which can be detected by the reactivity with PHA-L lectin, associates with increased metastatic potential. In human bladder carcinomas, reactivity with Lotus tetragonolobus agglutinin (LTA) correlates with increased metastatic potential. LTA appears to react with Lex structure [Gal beta 1--- 4(Fuc alpha 1----3)GlcNAc] in the cancer cells. Expression of sialilated dimeric Lex increases in the metastatic nests of human colon carcinomas. On the other hand, sialyl antigen MGl has the tendency to be expressed in human gastric adenocarcinoma of low metastatic potential. MGl is defined by a monoclonal antibody raised against Ricinus communis agglutinin receptors isolated from human gastric adenocarcinoma xenografted in nude mice. Continued efforts using monoclonal antibodies and lectins may yield arrays of carbohydrate markers of clinical value to predict the metastatic potential. PMID- 2572201 TI - [Cell cycle specific expression of the HER-2/NEU oncogene in breast cancer cell lines]. PMID- 2572200 TI - Increased contact hypersensitivity response in mice by topical application of 1 alpha,25-dihydroxyvitamin D3 to elicitation site. AB - Recent evidence indicates that the biologically active metabolite of vitamin D3, 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3], has an effect on the regulation of the immune response. We investigated whether topical treatment of mice with 1 alpha,25(OH)2D3 influences the contact hypersensitivity (CHS) response to trinitrochlorobenzene (TNCB). 1 alpha,25(OH)2D3 was applied to the dorsal trunk of A/J mice on days 0-3, and on day 4 topical application of 5% TNCB on the 1 alpha,25(OH)2D3-treated site was performed. The mice were tested for CHS on day 10 by applying 1% TNCB to the ears. No effect on induction of CHS response to TNCB was observed in 1 alpha,25(OH)2D3-treated mice compared with 24,25 dihydroxyvitamin D3[24,25(OH)2D3]-treated mice as control. In a second experiment, the dorsal trunk of A/J mice was treated with 5% TNCB on day 0. The topical application of 1 alpha,25(OH)2D3 on the ears was performed from days 2 to 5. On day 6, the mice were tested for CHS by applying 1% TNCB to the 1 alpha,25(OH)2D3-treated ears. When 1 alpha,25(OH)2D3 was administered to the elicitation site prior to the challenge, pretreatment of mice with 1 alpha,25(OH)2D3 increased their response to TNCB by 40% compared with 24,25(OH)2D3-treated mice as control (P less than 0.01). There were no findings suggesting that the pretreatment of the challenge site with 1 alpha,25(OH)2D3 induced an irritant dermatitis that was superimposed on a subsequent CHS reaction. The 1 alpha,25(OH)2D3 modulation of CHS response to TNCB in mice suggests that the hormone may play a role in the regulation of the immune response in vivo. PMID- 2572202 TI - [Oncogene organization and expression in breast cancer exemplified by proto oncogene c-erb B2]. PMID- 2572203 TI - [Immunoactive cells in endometrial tissue in various phases of the female ovulatory cycle]. PMID- 2572204 TI - Confusing messages from the newest of the beta-blocker/diuretic hypertension trials. The Metoprolol Atherosclerosis Prevention in Hypertensives trial. PMID- 2572205 TI - Treatment of tardive dyskinesia with bromocriptine. A test of the receptor modification strategy. AB - Pathophysiologic theories postulate that tardive dyskinesia arises from the development of chemical denervation supersensitivity of dopamine receptors produced by chronic long-term neuroleptic treatment. To test a dopamine receptor modification strategy, 16 patients with tardive dyskinesia were assigned to treatment with a neuroleptic plus bromocriptine (a dopamine agonist) or placebo for 10 weeks in a rising-dose design. Patients were evaluated weekly during the 10-week treatment period and for 8 weeks after medication withdrawal. No significant treatment effect was found in tardive dyskinesia response in the overall sample. When patients were classified by tardive dyskinesia subtype, patients with choreoathetoid symptoms exhibited only slight improvement with active treatment, which persisted after drug withdrawal; patients with dystonic symptoms showed moderate improvement that was drug and dose dependent. The sustained administration of substantial doses of a dopamine agonist did not produce significant adverse effects, including behavioral toxic effects. These results, although not statistically significant, are of clinical and heuristic interest. PMID- 2572206 TI - Risk factors for neuroleptic malignant syndrome. A case-control study. AB - A number of risk factors have been proposed for the development of neuroleptic malignant syndrome, but these have not been subjected to controlled study. To address this problem, we performed a case-control study comparing 18 patients with neuroleptic malignant syndrome and 36 matched neuroleptic-treated control patients with no known history of the syndrome to identify potential risk factors. Patients with neuroleptic malignant syndrome displayed significantly greater psychomotor agitation, received significantly higher doses of neuroleptics at greater rates of dosage increase, and received a greater number of intramuscular injections than controls. PMID- 2572207 TI - Antipsychotic drug use in the United States, 1976-1985. AB - Data from the National Prescription Audit and the National Disease and Therapeutic Index were used to assess trends in outpatient prescriptions for antipsychotic drugs in the United States from 1976 to 1985. Retail pharmacies dispensed 21 million prescriptions for antipsychotic drugs in 1976 and 19 million in 1985. The three leading antipsychotic drugs-thioridazine, haloperidol, and chlorpromazine-constituted 66% to 69% of antipsychotic prescriptions and, along with trifluoperazine, thiothixene, and fluphenazine, accounted for 90% to 91% of all antipsychotic prescriptions throughout the period studied. Thioridazine was the leading medication, with a consistent third of the market share, while the market share for chlorpromazine decreased from 1976 to 1985 and that for haloperidol increased for the same years. Data also indicate that haloperidol is the antipsychotic drug used most frequently in office-based, private medical practice for patients 60 years of age and older, a declining proportion of women are treated with antipsychotic medications, and use of antipsychotic drugs as monotherapy for the primary diagnosis is increasing. We also obtained data on diagnoses associated with antipsychotic drug use. PMID- 2572209 TI - Idazoxan: a selective alpha 2-antagonist and effective sustained antidepressant in two bipolar depressed patients. PMID- 2572208 TI - Effects of short-term administration of antipsychotic drugs on lymphocyte subsets in schizophrenic patients. PMID- 2572210 TI - [An unusual combination of primary-multiple apudomas and malignant angiomyolipoma of the kidney]. AB - A casuistic observation is described, in which 4 primary multiple tumors of various histogenesis were detected on autopsy in a 56-year-old patient. In the upper part of the jejunum, there was a 3 X 4 cm carcinoid of a mixed structure with metastases to the liver. The primary disseminated malignant glomas tumor as multiple interfluent nodes measuring up to 1.0 X 1.5 cm was present in the parietal and visceral peritoneum. A 2.0 X 2.5 x 3.0 cm pheochromocytoma and a 1.5 X 2.0 X 2.5 cm malignant angiomyolipoma were detected in the right kidney. The tumors had a typical histological structure, yet it was indicated that the detection of paragangliomoid sites in the malignant carcinoid slightly made it difficult to differentially diagnose it with the primary disseminated malignant glomus tumor of the peritoneum. PMID- 2572211 TI - The effect of sustained acceleration and noise on workload in human operators. AB - Biodynamic stressors such as acceleration, vibration, heat, and cold can affect pilot performance. The objective of this research was to determine the individual effect of two of these stressors, sustained acceleration or high intensity pink noise, on workload in human operators. Combined stressors were not investigated. A total of 13 workload measures, including 1 subjective, 4 performance, and 8 physiological, were recorded on subjects (N = 9) performing a dual psychomotor task in the human centrifuge. Increasing noise stress (max 100 dB-A weighted) had a significant effect on the Subjective Workload Assessment Technique or SWAT scores (p = 0.0001) and reaction times (p = 0.0189); acceleration stress (average peak, 3.75 Gz) had a significant effect on SWAT (p = 0.0001), heart rate (p = 0.0001), total eye blinks (p = 0.0184), blink duration (p = 0.0017), and the standard deviation of the EMG on the tracking forearm (p = 0.0452). Although many of the workload measures were affected by the noise or acceleration, subjects were able to maintain their performance on the primary tracking task. It is concluded that biodynamic stressors, such as noise and acceleration can adversely affect subjective operator workload without affecting objective task performance. PMID- 2572212 TI - Molecular defects of the C1-inhibitor gene in hereditary angioedema. AB - Family studies of hereditary angioedema are discussed with particular emphasis on the molecular basis of the more common form of the disease (type I), characterized by quantitative C1-Inhibitor deficiencies. Results obtained in several laboratories can be interpreted in the light of the restriction maps and of the exon-intron structure of the normal C1-Inhibitor gene that have been published recently. Several partial deletions of either exon 4 or exon 7 or of larger portions of the 5' end of the gene and a duplication of exon 4 have been described. These gene alterations are essentially family-specific and they account for at least fifteen-percent of the defects in type I hereditary angioedema. Detailed sequences analyses of cloned defective genes indicate that clusters of repetitive sequences of the Alu family, located within several introns, are responsible for the relatively high frequency of gross gene alterations observed at the C1-Inhibitor locus. In contrast, most of the defects that lead to production of dysfunctional proteins appear to be point mutations within the P1 residue of the reactive site, whose codon features a hypermutable CpG dinucleotide. PMID- 2572213 TI - Human complement C1r and C1s proteins and genes: studies with molecular probes. AB - The isolation of complementary DNA clones for both enzymic subcomponents of C1 has made it possible to derive their complete amino acid sequences and to verify and extend previous protein data. We review here recent advances in studies of the C1r and C1s proteins and of the corresponding genes, using molecular probes. The mosaic structure of these proteins has been compared to the exon-intron organization of the C1s gene. Surprisingly, the C1r and the C1s genes feature an intronless serine protease domain, at variance with all vertebrate serine proteases. Moreover, C1r and C1s are related in evolution to haptoglobin, a serine protease analog lacking enzymic activity. The C1r and C1s genes are closely linked in an unusual tail to tail orientation. These findings are discussed with regard to the apparently coordinate expression of these complement components and to the combined nature of most C1r and C1s deficiencies. We also discuss the implications of the successful production of C1r protein using recombinant DNA technology. PMID- 2572214 TI - Biochemical correlates of genetic variation in marine lower invertebrates. AB - In this paper extensive data on enzyme variation in 23 species of coelenterates and sponges were used to investigate the possible correlation of levels of genetic variation with various parameters of enzyme molecular structure and function. The data provide an opportunity not only to look for such correlations for the first time in lower invertebrates, but also to study organisms with far higher average levels of genetic variability than those used in any previous work. A clear inverse relationship was found between enzyme subunit number and levels of polymorphism, with monomers being more variable than dimers or tetramers. No significant difference in polymorphism could be found in enzymes of the functional groups I and II of Gillespie and Langley (1974). Regulatory enzymes appeared to be significant relationship was observed between regulatory power and subunit structure which could bias this result. The results suggest that both neutralist and selectionist ideas may have a useful role to play in the understanding of the factors which can influence or limit levels of genetic variation. PMID- 2572215 TI - DNA length variants contiguous to the 3' end of a vitellogenin gene in Drosophila grimshawi laboratory stocks from different Hawaiian Islands. AB - Two V3 vitellogenin clones isolated from genomic libraries of Drosophila grimshawi (G1, Auwahi, Maui) were found to differ in length. Structural comparison of the two clones established that the length difference could be attributed to two insertions/deletions of about 200 bp each, both within the 3' flanking sequences of the gene. The two length variants appeared to be polymorphic in the G1 laboratory strain, as demonstrated by analysis of genomic DNA isolated from single flies. The deleted variant sequence was traced by further analysis to two other D. grimshawi strains (PK9 and S10G1) which originated from the island of Molokai. The existence of this morph in the Maui strain appears to have resulted from a laboratory stock contamination at the Drosophila Stock Center. In the course of a few generations of culture of this G1 strain at New York University, the deleted morph increased its frequency surprisingly rapidly, almost replacing the original morph, while at the Bowling Green Stock Center, the original morph still predominates. These frequency changes are most likely consequences of genetic drift due to bottlenecks in the maintenance and propagation of this stock. PMID- 2572216 TI - The anomalous kinetics of sulphatase A. PMID- 2572218 TI - Oral mucosal pellicle. Adsorption and transpeptidation of salivary components to buccal epithelial cells. AB - The present investigation was carried out to examine the mechanism(s) whereby salivary molecules interact with human buccal epithelial cells. By utilizing antiserum against human parotid saliva, selected salivary components were detected by electrophoretic-transfer analysis of 1.5% SDS extracts of epithelial cells. Incubation of the cells and their aqueous cell-free extracts with 125I labelled parotid saliva resulted in the formation of an iodinated high-molecular mass complex which was not present in 125I-labelled saline alone. Formation of this complex was time-dependent and was inhibited by treating the buccal epithelial cells or their cell-free extracts with EGTA, iodoacetamide, N ethylmaleimide or by heating at 100 degrees C for 15 min. The epithelial cells also promoted incorporation of [14C]putrescine into high-molecular-mass complexes whose formation was inhibited by iodoacetamide, unlabelled putrescine and EGTA. Cell extracts mediated cross-linking of monodansylcadaverine into alpha-casein, and this interaction was inhibited by iodoacetamide. Significant amounts of radioactivity were recovered with the epithelial-cell envelopes after exhaustive extraction of 125I-saliva- or [14C]putrescine-treated epithelial cells with 4% (w/v) SDS/10% (v/v) beta-mercaptoethanol. The incorporation of radioactivity into epithelial-cell envelopes was inhibited by pretreatment of the cells with putrescine, EGTA, iodoacetamide, or heating at 100 degrees C for 15 min. These data suggest that: (1) oral mucosal pellicle is formed by the selective adsorption of saliva to the epithelial-cell plasma membrane and its associated cytoskeleton; and (2) the adsorbed salivary components may be cross-linked to each other or the epithelial cytoskeleton by epithelial transglutaminases. PMID- 2572217 TI - The novel thermogenic beta-adrenergic agonist Ro 16-8714 increases the interscapular brown-fat beta-receptor-adenylate cyclase and the uncoupling protein mRNA level in obese (fa/fa) Zucker rats. AB - In obese (fa/fa) rats both the total (-)-isoprenaline- and NaF-stimulated adenylate cyclase activities of interscapular brown-adipose-tissue (IBAT) plasma membranes were decreased as compared with lean rats by 40 and 38% respectively. An acute treatment with Ro 16-8714 increased (-)-isoprenaline- and NaF-stimulated adenylate cyclase activities by 2.5- and 2.0-fold respectively, and beta adrenoceptor number 2.8-fold in obese rat IBAT, but it had no effect on these parameters in lean rats. It increased mRNA for mitochondrial uncoupling protein in both lean and obese rats to the same extent. PMID- 2572219 TI - Biosynthesis of heparin. Relationship between the polymerization and sulphation processes. AB - Incubation of a mouse mastocytoma microsomal fraction with UDP-[3H]GlcA and UDP GlcNAc yielded proteoglycans containing non-sulphated polysaccharide chains. Similar incubations performed in the presence of sulphate donor 3' phosphoadenosine 5'-phosphosulphate (PAPS) produced both sulphated and non sulphated proteoglycans, which were separated by chromatography on DEAE-cellulose Analysis by gel chromatography of single polysaccharide chains, released from the proteoglycans by alkali treatment, showed that the non-sulphated chains produced during incubation for 5 min or 25 min, either in the absence or in the presence of PAPS, were of fairly small molecular size, with an average peak Mr of approx. 10 x 10(3)-15 x 10(3). In contrast, the sulphated chains exceeded Mr 100 x 10(3) Pulse-chase experiments suggested that sulphated chains were capable of further elongation. These results indicate that sulphation promotes, by so far unknown mechanisms, further chain elongation. Sulphated proteoglycan (retarded on DEAE cellulose chromatography) isolated after similar incubation of the microsomal fraction for 1 min only was found to contain a mixture of sulphated and virtually non-sulphated polysaccharide chains. However, when [35S]PAPS was included in the incubations, some 35S was found to be associated, essentially as N-sulphate groups, also with the latter type of chains, preferentially the high-Mr fraction. These results are interpreted in terms of a biosynthetic model by which the heparin proteoglycan is generated through transient interactions of macromolecular intermediates with distinctly separate complexes of membranebound enzymes. PMID- 2572220 TI - Electrophoretic mobility of gamma-glutamyltransferase in rat liver subcellular fractions. Evidence for structure difference from the kidney enzyme. AB - Adult rat liver gamma-glutamyltransferase (GGT) has been poorly characterized because of its very low concentration in the tissue. In contrast with the kidney, the liver enzyme is inducible by some xenobiotics, and its relationship to hepatic ontogeny and carcinogenesis seems to be important. Liver GGT polypeptides were identified by immunoblot analysis in subcellular fractions (rough endoplasmic reticulum, smooth endoplasmic reticulum, Golgi membranes and plasma membranes). Rat liver GGT appeared as a series of polypeptides corresponding to different maturation steps. Polypeptides related to the heavy subunit of GGT were detected in rough endoplasmic reticulum at 49, 53 and 55 kDa, and in Golgi membranes at 55, 60 and 66 kDa. Two polypeptides related to the light subunit of GGT were also observed in Golgi membranes. In plasma membranes GGT was composed of 100 kDa, 66 kDa and 31 kDa polypeptides. The 66 kDa component could correspond to the heavy subunit of the rat liver enzyme, and if so has a molecular mass higher than that of the purified rat kidney form of GGT (papain-treated). These data suggest different peptide backbones for the heavy subunits of liver GGT and kidney GGT. PMID- 2572222 TI - Influence of the placenta on fetal plasma insulin and neonatal hepatic enzyme induction in the rat. AB - In order to study the placental role in controlling glucose metabolism and neonatal induction of tyrosine aminotransferase in rat liver, a comparison was made between the artificially delivered-rat feto-placental unit and the newborn littermates. The immunoreactive serum insulin in the feto-placental unit remains as high as in utero for at least 1 h whereas in the newborn rat it decreases rapidly after delivery. From Caesarian section to 6 h later, in contrast to the newborn littermates, the feto-placental unit remains hyperglycaemic. Upon glucagon injection the feto-placental unit shows a delay in the increase of the hepatic tyrosine aminotransferase activity in comparison with the newborn rat. PMID- 2572221 TI - Bovine aortic endothelial cell transglutaminase. Enzyme characterization and regulation of activity. AB - Bovine aortic endothelial cells contain Ca2+-dependent tissue-type transglutaminase. Its activity in these cells was high, with apparent Km and Vmax. values with respect to putrescine of 0.203 mM and 18.5 nmol/min per mg of protein, and its activity was inhibited by the three competitive inhibitors dansylcadaverine, spermine and methylamine. The molecular mass of endothelial cell transglutaminase estimated by gel filtration chromatography was 88 kDa and it was immunoprecipitated by rabbit monospecific antiserum raised against rat liver transglutaminase. Its enzymic activity rose when the cell cultures reached confluence, and was further increased when their proliferation was arrested (synchronized at G0/G1 phase). Most of the enzymic activity was found in the 15,000 g soluble fraction, with only 4-22% of the activity found in the particulate fraction, depending on the state of cell proliferation. Examination of these cellular fractions by SDS/polyacrylamide-gel electrophoresis and immunoblotting revealed that at confluence endothelial cells have accumulated transglutaminase antigen in their 15,000 g particulate fraction. A series of experiments demonstrated the existence of a latent transglutaminase form in non proliferating cells, and suggested that this might involve the formation of an inhibitory complex. Treatment of cell lysates and the 15,000 g particulate fraction with high salt concentration showed a significant increase in transglutaminase activity. Mixing experiments using the 100,000 g particulate fraction or purified rat liver transglutaminase on one hand and the cytosolic fraction on the other showed dose-dependent inhibition of the transglutaminase activity of the latter. It is concluded that endothelial cells contain a particulate fraction-residing inhibitor of transglutaminase which interacts via ionic interaction with the enzyme. PMID- 2572223 TI - Increased membrane-associated transglutaminase activity in psoriasis. AB - Terminal differentiation of human skin involves the formation of an insoluble cross-linked protein envelope (CLE), which functions as an external barrier. To characterize terminal differentiation in the skin disease psoriasis, we have measured 1) membrane-associated transglutaminase (mTGase) activity, the rate limiting enzyme in the formation of CLE, and 2) the number of CLE in biopsies from normal and psoriatic skin. mTGase activity was increased 5-fold (p less than 0.0001) in psoriatic versus normal skin. Kinetic analysis revealed that the increased activity was due to an elevation in the Vmax of the enzyme. In addition, the number of CLE was 10-fold greater in psoriatic compared to normal skin. The increase in mTGase and CLE in psoriasis is in contrast to the decrease in other markers of terminal differentiation in skin, such as synthesis of specific intermediate filaments, observed in this disease. PMID- 2572224 TI - L-pipecolate oxidase: a distinct peroxisomal enzyme in man. AB - We investigated the oxidation of L-pipecolate in human liver. The results obtained with L-pipecolate from which traces of D-pipecolate had been removed by a preincubation with D-aminoacid oxidase indicate that a distinct L-pipecolate oxidase rather than D-aminoacid oxidase is responsible for the L-pipecolate dependent H2O2-production in human liver. Importantly, L-pipecolate oxidase was found to be localized in peroxisomes which adds to the growing number of enzymes and metabolic functions which can be ascribed to peroxisomes. PMID- 2572225 TI - DNA analysis of HLA-DR and DQ genes in American blacks with systemic lupus erythematosus. AB - We studied DNA polymorphisms of HLA-DR and DQ alleles in 63 American black patients with systemic lupus erythematosus (SLE). We found no HLA-DR beta, DQ alpha, or DQ beta restriction fragment length polymorphism (RFLP) or RFLP determined DR or DQ specificity associated with SLE in either the patients or in 57 control subjects. DRw52b was positively associated with renal involvement and negatively associated with anti-nuclear RNP antibodies. Antibodies to Ro (SS-A) and La (SS-B) were associated with DR3(DRw17), DQw2.3. Early-onset SLE (less than or equal to 20 years of age) was associated with DRw8, and the frequency of neuropsychiatric involvement correlated negatively with a 3.7-kb Taq I DQ alpha RFLP. This suggests a role for DR and DQ genes in the clinical and serologic expression of SLE in American blacks. PMID- 2572226 TI - Massive verapamil ingestion: a report of two cases and a review of the literature. AB - This report describes two patients who were victims of massive verapamil ingestion and then reviews the available literature. Because verapamil blocks the slow calcium channels of the heart and blood vessels, the use of calcium as a treatment would be logical. In the two cases reported here, calcium was only transiently effective in maintaining cardiac output and blood pressure. Several other agents were then used and most were ineffective. This is similar to experience reported in the literature that suggests that no single agent is capable of reversing verapamil's negative inotropic, dromotropic, chronotropic, and vascular smooth muscle effects. PMID- 2572227 TI - Beta-adrenoceptor desensitization in a model of experimental asthma in guinea pigs. AB - We investigated the possible occurrence of the desensitization of pulmonary beta receptors in a model of experimental asthma such as the ovalbumin sensitized guinea-pig. Although some differences were observed between normal and asthmatic guinea-pigs, the desensitization procedure (isoproterenol (ISO) 10(-6) M x 20 min x 2) markedly impaired the relaxing capacity of ISO, epinephrine (EPI) and procaterol. With the beta 2 agonist procaterol, the sensitized tissues seemed to be more sensitive compared to normal. In parallel, the desensitization procedure produced a greater decrease in the relaxing capacity of procaterol in the sensitized tracheas. On the other hand, the two unselective beta-agonists (ISO and EPI) did not seem to discriminate between the two experimental conditions used. These data suggest that the immunological disturbance due to the active sensitization may induce a modification in beta-adrenoceptor reactivity. The results obtained with EPI after anaphylactic challenge are in agreement with this. In fact, the relaxing capacity of EPI was markedly reduced by antigen challenge. PMID- 2572228 TI - Round table on antiasthmatic drugs; beta-agonists and theophylline. Workshop on pathogenesis and therapy of bronchial asthma Cortina d'Ampezzo/1-4 April, 1987. AB - Beta 2-agonists and theophylline compounds are first line drugs and are extensively used in the treatment of acute bronchial asthma. Beta 2-agonists are continuously expanding as a class of drugs. Recent research has increased the available knowledge of their cardiovascular and metabolic effects. Cardiovascular effects might be due to direct stimulation of beta 2-receptors sited in the atrial muscle. Metabolic effects (i.e. increase in blood glucose, insulin and non esterified fatty acid (NEFA), decrease in plasma potassium) are, at least partly, a consequence of direct or reflex stimulation. Despite recent progress, slow release theophylline compounds remain a basic approach to asthma management. Future progress in this field will probably be concerned with a more complete knowledge of 'ultra-slow' theophylline compounds (level of the plasma concentration fluctuation and patient compliance) and more complete results regarding the influence exerted by food (and gastric pH) on the rate and extent of absorption. PMID- 2572229 TI - Network regulation of autoimmunity: an automation model. AB - Four classes of regulatory T lymphocytes have been implicated in the control of experimental autoimmune diseases: a pair of helper and suppressor T lymphocytes that recognize the self-antigen (antigen-specific); and a pair of helper and suppressor T lymphocytes that recognize the autoimmune effector lymphocytes (anti idiotypic). The anti-idiotypic pair of regulators was detected following vaccination against autoimmune disease using autoimmune effector T clones as vaccines. To learn how the anti-idiotypic regulatory lymphocytes might function in concert with the antigen-specific regulatory lymphocytes, we devised a network in which the cell populations could be viewed as interconnected automata. Analysis of this novel network model suggests how self-tolerance may operate, how progressive autoimmune disease may develop, and how T-cell vaccination can control autoimmune disease. PMID- 2572230 TI - Anti-CD4 antibody treatment of patients with rheumatoid arthritis: I. Effect on clinical course and circulating T cells. AB - Eight patients with arthritis (seven with rheumatoid, one with psoriatic arthritis) were treated for 7 d with a daily injection of 10 mg of mouse monoclonal anti-CD4 antibodies (three with VIT4, five with MT151). With the exception of a short-lasting low-grade fever in one patient, no side effects were observed. Clinical symptoms (morning stiffness, number of swollen joints, pain assessment and Ritchie articular index) improved in all patients within 7 d of treatment. Improvement lasted from 3 weeks to greater than or equal to 5 months (mean approximately 11 weeks). Rheumatoid factors, immune complexes and other laboratory parameters did not change during or after treatment. Skin reactivity to recall antigens was suppressed in four out of six patients during treatment but returned to pretreatment levels within 6 weeks. Immunofluorescent analysis revealed a short-lasting drop of T cells, mainly of the CD4+ CDw29+ subset, but monocytes were also affected. The injected antibody was detectable on circulating cells for about 10 h. Within 20-24 h, the cell distribution returned to pretreatment levels. In six out of eight patients an anti-mouse-Ig response was seen. We conclude that mouse anti-CD4 monoclonal antibody (MoAb) treatment is well tolerated and that the cellular immunological changes observed are short lasting. The low incidence of side effects may justify further clinical studies to evaluate the clinical efficacy of such treatment. PMID- 2572231 TI - Anti-CD4 antibody treatment of patients with rheumatoid arthritis: II. Effect of in vivo treatment on in vitro proliferative response of CD4 cells. AB - In treating rheumatoid arthritis (RA) patients with anti-CD4 antibodies (MT151 10 mg/day, 7 d) we observed diminished skin-test response to recall antigens and reduced proliferative response to antigens and mitogens (in vitro) 2-4 h following the injection. To investigate whether this diminished response is due to functional impairment of CD4+ cells, we analyzed the proliferative response of CD4+ cells to various stimuli, in conditions where the CD4 structure was either cross-linked or not. We found that in vivo anti-CD4-antibody-coated cells could be induced to proliferate if the anti-CD4 antibody was cross-linked with an anti CD3 antibody [BMA030-F(ab')2] added in vitro and per se non-mitogenic. Also, without cross-linking the cell-bound anti-CD4 antibodies, no impairment of the proliferative capacity of CD4-antibody-coated cells could be detected; double fluorescence analyses of phytohaemagglutinin (PHA)- or CD3-stimulated cells showed that a similar proportion of CD4+ cells proliferated before and after anti CD4-antibody treatment. The diminished proliferation observed following anti-CD4 treatment correlated to the reduced number of CD3+ and CD4+ cells in culture, which suggests that altered cell distribution might be one factor contributing to the impaired immune response. PMID- 2572232 TI - Antipsychotic drugs, lithium, carbamazepine, and abnormal diurnal weight gain in psychosis. AB - Diurnal weight gain was abnormal among 149 institutionalized chronically psychotic patients. We weighed patients weekly for 3 weeks at 7 A.M. and 4 P.M. and then normalized the diurnal weight gain (NDWG) as a percentage by subtracting the 7 A.M. weight from the 4 P.M. weight, multiplying the difference by 100, and dividing the result by the 7 A.M. weight. The NDWG was 2.026% +/- 1.399% for patients and 0.511% +/- 0.351% for normals. The NDWG related (Spearman rho = 0.328, p less than 0.0001) to chlorpromazine equivalent dose. The absence of lithium and carbamazepine, lithium alone, carbamazepine alone, and the combination of lithium and carbamazepine did not alter NDWG. Our data suggest that antipsychotic drugs may contribute to abnormal weight gain in chronic psychosis but that lithium and carbamazepine do not. PMID- 2572233 TI - Activation of tuberoinfundibular dopamine neurons following the acute administration of atypical antipsychotics. AB - The activity of tuberoinfundibular dopamine neurons, as judged from dihydroxyphenylacetic acid (DOPAC) concentrations or the accumulation of dihydroxyphenylalanine (DOPA) in the median eminence after the inhibition of DOPA decarboxylase, was increased following the acute administration of the purported atypical antipsychotics clozapine, thioridazine, melperone, setoperone, and RMI 81582. In contrast, the activity of these hypothalamic dopamine neurons was not acutely altered by the typical antipsychotics haloperidol, chlorpromazine, fluphenazine, and cis-flupentixol or by SCH 23390. The acute stimulatory effect of the atypical antipsychotics on the activity of tuberoinfundibular dopamine neurons was effectively antagonized by the D1 agonist SKF 38393 but not by the D2 agonist quinpirole. The production of an acute activation of tuberoinfundibular dopamine neurons, which appears to be sensitive to D1 receptor activation, may be an effect that distinguishes typical and atypical antipyschotic agents. PMID- 2572234 TI - The carbachol-induced enhancement of desynchronized sleep signs is dose dependent and antagonized by centrally administered atropine. AB - Considerable data show that microinjection of carbachol into the pontine reticular formation produces a desynchronized (D) sleep-like state. The present study examined the hypothesis that this carbachol-induced enhancement of D sleep signs is mediated by muscarinic, cholinergic receptors. This hypothesis was tested by quantifying the dose-dependent effects of centrally administered carbachol on the D sleep-like state and by pretreating the animals with centrally administered atropine. Six dosages of carbachol were microinjected into the pontine reticular formation of conscious cats and polygraphic measures of behavioral state were recorded. The percentage, latency, duration, frequency, and time course of the carbachol-induced D sleep-like state were dose dependent. Centrally administered atropine competitively antagonized the ability of carbachol to induce the D sleep-like state, whereas pontine administration of L glutamate did not significantly alter D sleep. These data demonstrate that muscarinic, cholinergic receptors within the pontine reticular formation mediate the phenomenon of cholinoceptive D sleep sign enhancement. PMID- 2572235 TI - Enrichment of murine haemopoietic stem cells: diverging roads. AB - The cellular elements of the peripheral blood must be constantly replenished by the process of haemopoiesis, since most blood cells have a limited life span of only days or weeks. Although the developmental lineages of haemopoietic differentiation have been depicted in textbooks for decades, the actual details of the early stages of haemopoiesis are relatively unknown due to the very low numbers of haemopoietic stem cells in bone marrow or spleen. Only by isolating these rare stem cells and developing in-vitro culture systems to maintain them can a complete understanding of the early stages of haemopoiesis be achieved. This approach has already been successfully applied to the study of the later stages of haemopoiesis. In this review, Gerald Spangrude examines several experimental approaches that have been used to enrich murine haemopoietic stem cells. PMID- 2572236 TI - [Importance of Zyrtec in the treatment of chronic urticaria and allergic rhinopathies apropos of 1168 cases seen by hospital practitioners]. AB - This was a phase IV prospective trial involving out-patients from 80 hospital respiratory diseases and ENT departments, and 40 hospital dermatology departments. More than 1,000 cases were collected and used to evaluate the efficacy of Zyrtec at the dose of 10 mg per day. The aim of the trial was to assess the rapidity of action and good acceptability of treatment for 10 days and 2 weeks respectively in cases of seasonal and perennial rhinopathy. Similarly, in pruritic allergic skin disorders, the aim was to measure the degree of activity and acceptability of treatment for 2 weeks. Overall evaluation of results provided clinical evidence of satisfactory rapidity of action with an appreciable acceptability level. This prospective study confirms the good therapeutic index of Zyrtec as a new anti-allergic agent. PMID- 2572237 TI - Conformational effects of amino acid substitutions in the P-glycoprotein of the mdr 1 gene. AB - The P-glycoprotein of the mdr 1 gene is responsible for the phenomenon of multidrug resistance in human cells. The presumed drug-binding site of the wild type P-glycoprotein contains a glycine at position 185. A mutant P-glycoprotein which contains valine at this position causes cells to retain resistance to colchicine, but to lose cross-resistance to other drugs such as the chemotherapeutic agents vinblastine and Adriamycin. This has been hypothesized to be due to a conformational change in the protein induced by the amino acid substitution. Using conformational energy analysis, we have determined the allowed three-dimensional structures for the wild-type and mutant proteins in the region of position 185. The results indicate that the wild-type protein adopts a unique left-handed conformation at position 185 which is energically unfavorable for the protein with L-amino acids (including valine) at this position. This conformational change induced by amino acid substitutions for Gly 185 could explain the differences in binding to the P-glycoprotein of various drugs and, hence, the differences in drug resistance exhibited by various cell lines expressing these proteins. PMID- 2572239 TI - Paternity testing. AB - There is evidence that parentage determination was a problem even in biblical times. The most significant developments in resolving these problems occurred in the 20th century starting with Landsteiner's description of the ABO blood group system in 1900. The utility of paternity testing increased as new blood group systems were identified. Major advances occurred between 1940 and 1970, starting with the discovery of the Rh system and reaching a highpoint with the application of HLA typing to paternity studies. Although the exclusion of falsely accused alleged fathers continues as the primary focus of paternity testing, statistical analysis has contributed significantly to the information that can be gleaned from the test results when exclusion has not been demonstrated. Paternal inclusion still cannot be conclusively proven but a mathematical estimation of the likelihood of paternity has found legal acceptance. New technology is now being directed toward identifying inherited DNA markers through the use of genetic probes. Segregation of restriction fragment length polymorphisms (RFLP) may provide the near ultimate test for paternity as it may supply indisputable evidence of family relationships. PMID- 2572238 TI - Interleukin-1 beta gene deregulation associated with chromosomal rearrangement: a candidate initiating event for murine radiation-myeloid leukemogenesis? AB - The incidence of acute myeloid leukemia (AML) in CBA/H mice following exposure to single acute doses of ionizing radiation has previously been determined. A high proportion of these AMLs are characterized by rearrangement of murine chromosome 2 in the C2 and/or E5-F regions, and there is evidence that these events are a direct consequence of radiation damage to multipotential hemopoietic cells. Using a combination of in situ chromosome hybridization and mRNA analyses, we show that the cytokine gene interleukin-1 beta (IL-1 beta) is encoded in the chromosome 2 F region and is translocated in a chromosome 2---2 rearrangement in an x-ray induced AML (N36). Also, IL-1 beta is specifically deregulated in N36 and in two other chromosome 2-rearranged AMLs but not in a fourth, which has two cytogenetically normal chromosome 2 copies. We suggest that radiation-induced specific chromosome 2 rearrangement associated with IL-1 beta deregulation may initiate murine leukemogenesis through the uncoupling of normal proliferative control mechanisms in multipotential hemopoietic cells. PMID- 2572240 TI - Biology of human imunodeficiency virus: virus receptor and cell tropism. PMID- 2572241 TI - Convergence of hippocampal and dopaminergic input onto identified neurons in the nucleus accumbens of the rat. AB - The hippocampal input to the nucleus accumbens was interrupted by an electrolytic lesion of the fimbria-fornix. Boutons degenerating as a result of this lesion were found in asymmetric synaptic contact with dendritic spines and shafts in the medial part of the nucleus accumbens. Dopaminergic fibres and terminals in this area, identified using an antibody to tyrosine hydroxylase, established symmetrical synaptic contacts with dendritic shafts, spines and somata. In material where neurons in the nucleus accumbens had been Golgi-impregnated, it was found that the hippocampal and dopaminergic inputs converge onto the same neurons, and that the post-synaptic targets could be either spiny or aspiny neurons. It has been suggested that hippocampal dysfunction is involved in schizophrenia and this convergence of input from the hippocampus onto the same neurons that are post-synaptic to the dopaminergic input, which presumably originates from neurons in the ventral tegmental area, may provide an anatomical basis for the therapeutic effects of neuroleptic drugs which are dopamine antagonists. PMID- 2572242 TI - Angiotensin II-producing enzyme III from acidified serum of nephrectomized dogs. AB - A highly active angiotensin-producing enzyme (enzyme III) was obtained from the serum of bilaterally nephrectomized dogs by acid treatment and ammonium sulfate fractionation. An inactive precursor (proenzyme III) was converted to enzyme III during prolonged storage (or by treatment with acid or with cathepsin G or by incubation at 38 degrees C as described in the following paper). Enzyme III reacted maximally at pH 7.7 and it produced up to 400 ng of angiotensin II/mL serum/h (i.e., amounts 4000 times higher than that generated by the endogenous renin present in serum after bilateral nephrectomy). Enzyme III produced angiotensin II at identical rates when either dog angiotensinogen or angiotensin I was used as substrate, but the rate was 710 times higher with synthetic tetradecapeptide renin substrate. Enzyme III is not identical to renin, cathepsin G, tonin, enzyme I, enzyme II, the calcium-dependent angiotensin I-converting enzyme, or the calcium-independent carboxy peptidase, which acts by sequential cleavage of angiotensin I. Enzyme III was inhibited by alpha-1-antitrypsin, diisopropyl fluorophosphate, and lima bean trypsin inhibitor (hence it is a serine proteinase). It was not inhibited by Captopril, Teprotide, or Enalapril. It had been reported previously that cathepsin G released from neutrophil granulocytes, by producing high local concentrations of angiotensin II, may provide a mobile means for modulating blood flow in tissue microvasculature during the inflammatory response. The present study offers a new, additional pathway, by enzyme III, for a similar rapid formation of angiotensin II from serum protein substrate or angiotensin I. PMID- 2572243 TI - New perspectives on selective alpha 1 blockade. AB - Selective alpha 1 adrenergic receptor blocking agents lower blood pressure by reducing the increased peripheral vascular resistance that characterizes essential hypertension. Prazosin and terazosin have been shown to be well tolerated in clinical practice and seldom cause impotence or metabolic abnormalities. The most common adverse effects--dizziness, headache, and asthenia -are generally well tolerated and infrequently lead to discontinuation of therapy. First-dose syncope can usually be avoided by initiating therapy with low doses administered at bedtime. Finally, the alpha 1 receptor antagonists do not adversely affect such cardiovascular risk factors as hypokalemia, serum lipid profile, and left ventricular hypertrophy. In fact, alpha 1 antagonists reduce total cholesterol and low-density-lipoprotein plus very-low-density-lipoprotein cholesterol and thus may contribute to the overall management of cardiovascular risk by blood pressure reduction and improvement of the serum lipid profile. Since the goal of treating chronic essential hypertension is to improve morbidity and mortality, the choice of therapy should be influenced by the agent's ability to modify as many risk factors as possible. Alpha 1 adrenoreceptor antagonists beneficially impact several cardiovascular risk factors and thus merit consideration as first-line antihypertensive therapy. PMID- 2572244 TI - Excitatory sulphur amino acid-evoked neurotransmitter release from rat brain synaptosome fractions. AB - The neuroactive sulphur-containing amino acids L-cysteate (CA), L-cysteine sulphinate (CSA), L-homocysteine sulphinate (HSA), S-sulpho-L-cysteine (SC) and L homocysteate (HCA) evoked the release of previously accumulated D-[3H]aspartate from rat brain cerebrocortical and cerebellar synaptosome fractions in a manner that was wholly Ca2+-independent. However, analysis of endogenous release by hplc revealed the presence of both Ca2+-dependent and -independent component of L glutamate release but only a Ca2+-independent component of L-aspartate release. CA, CSA, HSA and SC but not HCA evoked the release of previously accumulated [3H]GABA from synaptosome fractions by a mechanism shown to comprise both a Ca2+ dependent and -independent component. The specific antagonists of the N-methyl-D aspartate (NMDA) receptor, 3-[(+/-)-2-carboxypiperazin-4-yl]propyl-l-phosphonic acid (CPP) and the relatively selective competitive quisqualate (QUIS)/kainate (KA) receptor antagonist, 6-cyano-7-dinitroquinoxalinedione (CNQX), were ineffective in blocking the excitatory sulphur amino acid-evoked release of either D-[3H]aspartate, [3H]GABA or of endogenous established transmitter amino acids. PMID- 2572245 TI - Influence of cysteamine and cysteine on open-field behaviour, and on brain concentrations of catecholamines, somatostatin, neuropeptide Y, and corticotropin releasing hormone in the rat. AB - Cysteamine (1.95 or 3.90 mM/kg) administered subcutaneously (sc) markedly decreased the open-field activity of the rats, while the structurally related amino acid cysteine had only minor influence. Cysteamine (1.95 or 3.90 mM/kg) reduced the noradrenaline and increased the dopamine and dihydroxyphenyl acetic acid (DOPAC) levels in the hypothalamus. In striatum the drug decreased both the noradrenaline (1.95 or 3.90 mM/kg) and dopamine (3.90 mM/kg) levels without influencing the DOPAC content. Neither the hypothalamic nor the striatal catecholamines are influenced by administration of equimolar doses of cysteine. Cysteamine (1.95 or 3.90 mM/kg) decreased the somatostatin levels both in the hypothalamus and in the striatum without influencing neuropeptide Y (NPY) and corticotropin releasing hormone (CRH) concentrations. Cysteine administered in equimolar doses did not influence the peptide levels in these brain structures. These data suggest that the cysteamine-induced behavioural changes are related to the decrease of brain noradrenaline and somatostatin concentrations. The structurally related amino acid cysteine does not influence the behaviour or the central monoaminergic and peptidergic concentrations in the hypothalamus and striatum of rats. PMID- 2572246 TI - [Spinal subdural empyema diagnosed by MRI and recovered by conservative treatment]. AB - Localized suppuration involving the spinal cord is uncommon. A case of spinal subdural empyema is reported. The patient is 54-year-old male who had been suffering a diabetes mellitus but did not receive any treatment. His initial symptom was lumbago. Then he noticed a palpitation and general malaise which made him visit a hospital. Because he did not show any improvement by a fluid therapy, he was transferred to our institute for the further evaluation. On admission, physical examination showed no abnormality. Blood pressure was 170/90 mmHg, heart rate 128/min. and body temperature 37.1 degrees C suggesting a septic shock state. Neurological examination revealed slight consciousness disturbance, mild tetraparesis and bilateral hypesthesia lower than the level of L3. Laboratory examination showed the elevated leukocyte count and fasting blood sugar and urine ketone body levels of 20,500/mm3, 257 mg/dl and 226 mg/dl respectively. Blood culture proved a septicemia of Streptococcus agalactiae afterwards. On the second day of admission, lumbar puncture revealed a purulent cerebrospinal fluid, though X-ray CT of lumbar spine did not confirm a diagnosis. Spinal magnetic resonance imaging (MRI) revealed a widespread abnormal intensity of the spinal canal from the level of Th11 to L4. On the T1-weighted image (TR 300 msec., TE 40 msec.), cerebrospinal fluid space was abnormally isointense. On the T2-weighted image (TR 2,000 msec., TE 80 msec.), subdural and cerebrospinal space was filled with an abnormal high-intense lesion especially on the ventral side. He developed semicoma due to hydrocephalus following a intraventricular empyema. He was also complicated disseminated intravascular coagulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572248 TI - Neuromuscular block by suxamethonium following treatment with histamine type 2 antagonists or metoclopramide. AB - We have examined the effect of preoperative i.v. administration of three different histamine type 2 (H2) antagonists (cimetidine 400 mg, ranitidine 80 mg and famotidine 20 mg) or metoclopramide 10 mg i.v. on the duration of neuromuscular block produced by an intubating dose (1 mg kg-1) of suxamethonium. The study was conducted blindly in 70 post partum patients, weighing between 45 and 120 kg, scheduled for tubal ligation. The duration of neuromuscular block following suxamethonium was measured from a chart recording of train-of-four (TOF) electromyographic responses. The time from onset of 95% block to 25% recovery ("block time") was not significantly different between the groups receiving cimetidine (A), ranitidine (B), famotidine (C), and control (E). However, there was prolongation of neuromuscular block in patients receiving suxamethonium and metoclopramide (D). PMID- 2572247 TI - Enflurane-induced burst discharge of hippocampal CA1 neurones is blocked by the NMDA receptor antagonist APV. AB - The basis for the hyperexcitability and seizure activity associated with enflurane anaesthesia was investigated using extracellular and intracellular recording in rat hippocampal brain slices. Enflurane produced seizure-like burst discharges in CA1 pyramidal neurones, accompanied by depressed field potential amplitudes and a reduced threshold for synaptically evoked population spikes. However, threshold for action potentials evoked by intracellular current injection did not change, nor did action potential amplitude, duration or spike frequency accommodation in single neurones. Enflurane 2.0 vol% hyperpolarized CA1 neurones (3.1 (SD 1.3)mV), decreased membrane conductance (12 (6)% below control), and depressed EPSP amplitudes (34% of control) (P less than 0.01). Enflurane appeared to enhance both intrinsic and synaptically mediated inhibitory potentials. The N-methyl-D-aspartate (NMDA) receptor antagonist amino phosphonovalerate (APV) 5-20 mumol litre-1 completely blocked seizure-like burst discharge of CA1 neurones in the presence of enflurane, and the enflurane-induced reduction of population spike threshold; it did not alter anaesthetic depression of EPSP amplitude. Thus enflurane-induced burst discharge of CA1 neurones appeared to involve an enhancement of excitatory synaptic transmission rather than depression of intrinsic or synaptic inhibition. PMID- 2572249 TI - Xamoterol, a beta1-adrenoceptor partial agonist: a new approach to heart failure. Proceedings of the 1st International Heart Failure. Interlaken, 1988. PMID- 2572250 TI - Effects of xamoterol on resting and exercise haemodynamics in patients with chronic heart failure. AB - 1. A bicycle exercise test was used to investigate functional capability and haemodynamics in 30 patients with heart failure (13 NYHA Class II, 17 Class III), before and after i.v. xamoterol (Corwin, Carwin, Corwil, Xamtol, ICI 118,587) 0.2 mg kg-1. 2. Resting heart rate fell from 78 to 74 beats min-1 (P less than 0.05) and cardiac index rose from 2.5 to 2.8 l min-1 m-2 (P less than 0.001) after xamoterol. Blood pressure fell slightly, and systemic vascular resistance was reduced. Stroke work index improved and double product decreased. There were no changes in pulmonary artery wedge pressure ejection fraction or plasma noradrenaline concentrations. 3. On exercise, xamoterol produced a considerable reduction in heart rate increase, improved stroke volume and left ventricular stroke index and lowered double product. Exercise duration increased by 10%, but this did not quite achieve statistical significance. 4. These results are consistent with the concept that a beta 1-partial adrenoceptor agonist with the level of intrinsic sympathomimetic activity (43%) of xamoterol provides moderate inotropic support at rest, and protects the heart against overstimulation on exercise, when sympathetic drive is high. 5. Reduction of double product on exercise implies a lowered oxygen demand, which could be of considerable importance in patients with ischaemic heart disease. PMID- 2572251 TI - Xamoterol, a beta 1-adrenoceptor partial agonist: review of the clinical efficacy in heart failure. AB - 1. Xamoterol (Corwin, Carwin, Corwil, Xamtol, ICI 118,587), a beta 1-adrenoceptor partial agonist, improves both systolic and diastolic function in heart failure patients. 2. Double-blind, randomised studies comparing xamoterol with placebo showed that the beneficial haemodynamic effects of xamoterol produced significant improvements in exercise capacity and symptoms in patients with mild to moderate heart failure. These studies formed the basis for a large European multicentre study programme which recruited over 1000 patients, randomised to xamoterol (200 mg twice daily, n = 617), digoxin (0.125 mg twice daily, n = 135) or placebo (n = 300) for 3 months. 3. Efficacy was assessed by measuring exercise capacity and symptoms. The xamoterol group improved exercise capacity by 37% compared with an 18% improvement in the placebo group. Differences in the symptom scores measured by visual analogue scales and Likert scores indicated significant improvements by xamoterol in the cardinal symptoms of heart failure, dyspnoea and fatigue. 4. Analyses of data from subsets of patients in the study showed that elderly patients, patients on no other heart failure therapy and patients with cardiomegaly all had similar improvements in exercise and symptoms to those seen in the whole study population. In the subset which included digoxin treatment, xamoterol produced significantly greater improvements in exercise capacity than digoxin (33% vs 17%, P less than 0.05) and was associated with fewer side effects. 5. Xamoterol is therefore a promising addition to heart failure therapies currently available. PMID- 2572252 TI - The pharmacology of xamoterol: a basis for modulation of the autonomic control of the heart. AB - 1. Xamoterol (Corwin, Carwin, Corwil, Xamtol, ICI 118,587) is a beta-adrenoceptor partial agonist which is of benefit in the chronic treatment of heart failure (e.g. The German and Austrian Xamoterol Study Group, 1988). These results contrast with those obtained with other beta-adrenoceptor drugs, prenalterol and pirbuterol which were unsuccessful on chronic dosing (Currie et al., 1984; Glover et al., 1985). 2. Unlike prenalterol and pirbuterol, xamoterol has no significant agonist activity at the beta 2-adrenoceptor and has shown no tachyphylaxis in animals or man. 3. The overall action of xamoterol is to modulate sympathetic control of the heart such that at rest and at low levels of exercise the heart receives inotropic support whilst during more severe exercise, heart rate is reduced. In patients with left ventricular dysfunction these effects lead to an improvement in the relation between filling pressure and cardiac output at all levels of activity such that a given cardiac output is achieved with a lower filling pressure. It is suggested that this alteration in the pattern of cardiac activity over a long period of time results in a beneficial adaptation of the myocardium and is a possible explanation of the observed clinical improvement. PMID- 2572253 TI - Treatment of congestive heart failure--state of the art and future trends. AB - 1. Cardiac failure is a clinical syndrome of symptoms and signs, which can be confirmed by imaging or invasive haemodynamic techniques. It may be caused by systolic or diastolic dysfunction, but systolic dysfunction rarely occurs alone. It is important to ascertain the degree to which each contributes, and the precise aetiology of the condition, particularly in relation to surgically correctable lesions. 2. Non-pharmacological approaches including weight loss, salt restriction and lifestyle changes may be beneficial in some patients, and diuretics, which reduce the load on the heart, are the traditional baseline therapy. 3. Digitalis has been used where problems with contractility predominate, but its beneficial effect has been disputed, and expectations of improvement in patients in sinus rhythm should not be too high. 4. Vasodilators have been considered as the next line of treatment. Arteriolar dilators tend to increase cardiac output, but have little effect on pulmonary artery wedge pressure, and venodilators tend to have the opposite effect. Probably both actions are necessary and angiotensin converting enzyme (ACE) inhibitors, which have both, have proved effective in terms of symptoms and survival. 5. Various other inotropic agents have been tried. Phosphodiesterase inhibitors improve exercise tolerance, but may increase the probability of serious arrhythmias, already a significant cause of sudden death. beta 1-partial adrenoceptor agonists such as xamoterol have shown some promise, and anti-arrhythmic therapy has also been considered. 6. Drugs which prevent progression of myocardial damage would prove a great advance, and beta-adrenoceptor antagonists and calcium channel blockers appear to have considerable potential in this area. PMID- 2572255 TI - Long-term studies with xamoterol in heart failure. AB - 1. Xamoterol (Corwin, Carwin, Corwil, Xamtol, ICI 118,587) is a beta 1 adrenoceptor partial agonist which, unlike full beta-adrenoceptor agonists, does not down-regulate beta-adrenoceptors in the rat ventricle during chronic administration. 2. Improvements in myocardial performance have been demonstrated following acute administration of xamoterol to patients with mild or moderate heart failure, and these are sustained during at least 1 year of continued treatment. 3. Exercise duration is increased by xamoterol in patients with left ventricular dysfunction and benefit is still apparent after at least 1 year of therapy. 4. Despite sustained cardiac stimulation, xamoterol does not appear to affect adversely mortality in patients with mild or moderate heart failure. 5. Few adverse events directly attributable to xamoterol were reported during 3 month efficacy studies in more than 600 patients, and the laboratory safety profile over 1 year of treatment is good. 6. Xamoterol is a promising, well tolerated addition to established therapies for chronic mild or moderate heart failure. PMID- 2572254 TI - Focus on diastolic dysfunction: a new approach to heart failure therapy. AB - 1. Although heart failure is commonly associated with depressed systolic function, there is increasing evidence that impaired diastolic performance is also universally present and might be a key determinant of symptoms, physical capacity and even survival in some subsets of patients. 2. Reduced diastolic distensibility increases cardiac filling pressure not only at rest, but even more during exercise when diastolic filling time is reduced. The increases in filling pressure and diastolic wall stress lead to pulmonary congestion and subendocardial ischaemia, it also triggers myocardial hypertrophy and a detrimental remodelling of the ventricular cavity. Perhaps even more importantly, impaired ventricular distensibility limits the use of the Frank-Starling mechanism, impairing systolic pump function and cardiac output adaptation during exercise. Therapies able to improve the distensibility of the ventricle are, therefore, desirable in heart failure. 3. Nitrates, angiotensin converting enzyme (ACE) inhibitors and diuretics may indirectly increase left ventricular chamber compliance by their effects on the right side of the heart. Cardiac glycosides do not improve myocardial relaxation and may even cause diastolic contracture at toxic doses. The new beta 1-adrenoceptor partial agonist, xamoterol, on the other hand, consistently lowers left ventricular filling pressure at rest and during exercise, and produces an increase in left ventricular dynamic compliance through the direct lusitropic effect of beta 1-adrenoceptor stimulation. These beneficial effects are maintained during prolonged therapy and also appear sufficient to slow the remodelling of the ventricular cavity. The improvement in symptoms and in exercise tolerance observed during xamoterol (Corwin, Carwin, Corwil, Xamtol, ICI 118,587) therapy might, therefore, be related to the improvement in left ventricular diastolic distensibility induced by this drug. PMID- 2572256 TI - The management of heart failure and the scope for new therapies: what role for xamoterol? AB - 1. Current therapy of heart failure leaves much to be desired. Not all patients respond, and many agents lose their effects with time. 2. Newer agents may be effective but toxic, and some which have a beneficial action when given intravenously have proved disappointing when used orally. 3. The value of digoxin in patients in sinus rhythm is open to debate, and diuretics, although useful acutely in reducing fluid overload, do not appear to improve prognosis. 4. Vasodilators increase effort capacity and reduce symptoms, possibly conferring some long-term benefit, and angiotensin converting enzyme (ACE) inhibitors improve symptoms and decrease mortality in a wide range of patients. 5. Positive inotropes may be effective in the short term, but they increase myocardial oxygen demand and show tachyphylaxis with no prognostic benefit. 6. Xamoterol (Corwin, Carwin, Corwil, Xamtol, ICI 118,587) is a partial sympathetic agonist with approximately 50% of the activity of a pure agonist, which provides inotropic support at rest, and protection against excess sympathetic activity on exercise. 7. It is compatible with other therapies and has shown no serious toxicity. 8. It should be considered at present as an adjunct to diuretic and/or ACE inhibitor therapy, although it may be useful alone; its role will become clearer as its effects on mortality are established. PMID- 2572257 TI - The effect of age and renal impairment on the pharmacokinetics of xamoterol. AB - The effects of age and renal function on the pharmacokinetics and plasma concentrations of xamoterol were examined. Peak and steady state concentrations were higher in older patients, but this was due to deterioration of renal function rather than to age itself. The dose of xamoterol only need be reduced in the elderly if there is evidence of impaired renal function. PMID- 2572258 TI - Xamoterol in mild to moderate heart failure: a subgroup analysis of patients with cardiomegaly but no concomitant angina pectoris. AB - Xamoterol has been shown in large, double-blind studies to produce benefit in patients with heart failure. Ischaemic heart disease is the commonest cause of heart failure in the Western World and many patients with heart failure also have angina pectoris (Califf et al., 1982). In view of the known anti-ischaemic effects of xamoterol, we analysed the results of a subgroup of 269 patients with heart failure but without chest pain as a limiting factor on exercise to compare the efficacy of xamoterol in such patients with that of the total group. There were no differences in exercise heart rate, exercise tolerance and symptoms in patients without chest pain compared with the total group. Xamoterol is probably, therefore, acting through myocardial mechanisms other than an anti-ischaemic effect. PMID- 2572259 TI - Effect of xamoterol on peripheral blood vessels in healthy subjects. AB - Changes in peripheral blood flow in response to increasing doses of xamoterol with or without a beta 2-adrenoceptor blocking agent were assessed in eight healthy volunteers. Xamoterol produced increases in femoral artery diameter, blood volume flow and velocity flow. In the brachial and posterior tibial arteries, systolic blood pressure rose, and in the brachial artery, diastolic blood pressure fell. None of these effects was affected by beta 2-adrenoceptor blockade. The actions of xamoterol on the circulation are consistent with direct stimulation of myocardial beta 1-adrenoceptors. PMID- 2572261 TI - The effects of age, sex and treatment with xamoterol on exercise capacity in patients with heart failure. AB - The influence of age, sex and xamoterol treatment on exercise capacity has been investigated in 705 heart failure patients who took part in a multicentre, placebo-controlled study. Regression analysis suggests that although less than 20% of the total variation in exercise capacity between patients is explained by these factors, useful information may be gained by taking them into account. Exercise capacity declines with age, more rapidly in males than in females; the benefits of xamoterol are independent of age and sex. This approach demonstrates that exercise testing is a sensitive indicator of physical performance and allows the effects of treatment to be seen in context. PMID- 2572260 TI - Effect of beta-adrenoceptor agents on the blood flow in carotid arteries in normal volunteers. AB - The effects on carotid artery blood flow of atenolol (a selective beta 1 adrenoceptor antagonist) and xamoterol (a beta 1-adrenoceptor partial agonist with 43% agonist activity) were measured using a Doppler technique in eight healthy volunteers. Atenolol produced a decrease in blood pressure and blood velocity flow, and tended to reduce volume flow. In contrast, there were no changes with xamoterol or placebo. Beta 1-adrenoceptor blockade may reduce cerebral blood flow but the partial agonist xamoterol does not appear to reduce carotid blood flow. PMID- 2572262 TI - The cardiovascular pharmacology of xamoterol, cicloprolol, prenalterol and pindolol in the anaesthetised dog. AB - An anaesthetised dog preparation was used to examine the pharmacological profiles of xamoterol, cicloprolol, prenalterol and pindolol. The effects of these agents on heart rate and hind limb perfusion pressure revealed that xamoterol is highly cardioselective and has no significant agonist activity at beta 2-receptors and no membrane stabilising effect. These pharmacological properties are not complicated by the formation of active metabolites and may explain the clinical benefits in the long-term treatment of heart failure. PMID- 2572263 TI - The efficacy and safety of chronic oral administration of xamoterol to patients with severe heart failure treated with ACE inhibitors. AB - Xamoterol 200 mg twice daily was given for 2 months to nine patients with severe heart failure already being treated with angiotensin converting enzyme (ACE) inhibitors. Left ventricular end-diastolic pressure fell from 28 to 13 mmHg and end-systolic volume fell from 115 to 106 ml m-2; indices of contractility improved and ejection fraction rose from 33 to 38%. The time constant of ventricular relaxation, T1, improved from 62 to 44 ms. Exercise tolerance improved. Thus, in this group of patients with severe heart failure, xamoterol produced benefits in systolic and diastolic function. There were no adverse effects. PMID- 2572264 TI - Contrasting effects of single doses of pindolol and xamoterol on left ventricular diastolic function. AB - Impaired left ventricular diastolic dysfunction is common in patients with ischaemic heart failure. The beta 1-adrenoceptor partial agonist xamoterol was compared with pindolol, a beta-adrenoceptor blocker with intrinsic sympathomimetic activity, in a haemodynamic study in 17 patients with ischaemic heart disease. Pindolol caused left ventricular end-diastolic pressure, wall stress and T1 to rise, whereas xamoterol produced improvements in all three parameters. These results suggest that xamoterol may be of greater benefit to patients in heart failure. PMID- 2572265 TI - Long-term efficacy of xamoterol (a beta 1-adrenoceptor partial agonist) in patients with mild to moderate heart failure. AB - This study was designed to assess the efficacy of xamoterol in 14 patients with mild to moderate heart failure over a period of 18 months. A beneficial effect on exercise capacity and a lowering of heart rate on exercise was sustained, and ejection fraction did not change, although some deterioration may be expected over this length of time in patients with heart failure. Xamoterol is safe and effective, and its benefits are maintained over at least 18 months. PMID- 2572266 TI - Effects of xamoterol on sodium excretion in volunteers. AB - Xamoterol has been shown to reduce the frequency of oedema and lung crepitations in heart failure. We examined its effects on blood pressure and renal function in healthy volunteers. Systolic blood pressure rose, sodium and chloride excretion increased and there was a strong correlation in individual subjects between rises in systolic blood pressure and in sodium excretion. Although no changes in glomerular filtration rates were seen, changes sufficient to explain the observed rise in sodium excretion are well within the experimental error of this study. Xamoterol may increase sodium excretion by an action on renal haemodynamics. PMID- 2572267 TI - Pattern of expression of c-erbB-2 oncoprotein in human fetuses. AB - The pattern of expression of the c-erbB-2 oncoprotein was investigated in whole mount preparations of 11 human fetuses by immunocytochemistry using two polyclonal antibodies, 20N and 21N. c-erbB-2 was widely expressed within all three germ layers. Expression remained relatively constant in epithelial, mesodermal and extraembryonic tissues, but varied over time during the development of the fetal skeleton. Western blotting failed to detect c-erbB-2 in normal fetal tissues but did confirm expression in a microvillous membrane preparation of placenta. c-erbB-2 expression is widespread in the human fetus and occurs at an earlier stage than epidermal growth factor receptor. PMID- 2572269 TI - Development of an ELISA to detect early local relapse of colorectal cancer. AB - The Y haptenic blood group determinant is expressed on the cells of malignant gastrointestinal tumours and on the normal gastrointestinal epithelium of individuals who can secrete blood group antigens. an enzyme-linked immunosorbent assay incorporating the high affinity biotin-avidin interaction was developed to measure the serum levels of antigens bearing the Y hapten in patients with colorectal cancer. The specificity of the assays was between 88 and 93% and the sensitivity in detecting extensive disease was between 24 and 34%. However, up to 67% of patients with local or abdominal recurrent disease secreted antigens expressing the Y hapten, whereas only 30% of patients with overt hepatic metastases secreted a similar antigen. Recognition of antigens bearing the Y hapten may therefore be useful in detecting early local relapse of colorectal cancer when a second operation to excise the recurrence may be possible. PMID- 2572268 TI - c-erbB-2/c-erbA co-amplification indicative of lymph node metastasis, and c-myc amplification of high tumour grade, in human breast carcinoma. AB - A panel of 73 samples, including 52 primary breast carcinomas, 10 normal breast tissues and 11 axillary lymph nodes, has been analysed for the presence of amplifications and gross structural alterations, in the oncogenes c-erbB-2, c erbA, c-myc, N-myc, c-mos and c-Ha-ras. The tumours were also classified, graded and staged histopathologically and their DNA ploidy (42 samples) was determined by flow cytometry. Three breast cancer cell lines (MCF7, ZR-75-1 and T47D) were also included in the study. Amplification of c-erbB-2 was detected in 28% of the tumours, of which 91% had an increased steady-state level of c-erbB-2 mRNA. Amplification of c-erbA was found in 23% of tumours and was always associated with the amplification of c-erbB-2. Ten out of 12 (83%) tumours which had c-erbB 2 and c-erbA co-amplification had metastasised to axillary lymph nodes (P less than 0.006). However, the human thymidine kinase gene, which is present at the same chromosomal location as these two oncogenes (17q21-22), was amplified in only tw tumours. Amplification of c-myc was detected in 21% of the tumours studied, of which 82% (P less than 0.005) were of histopathological grade 3 and none were of grade 1. Flow cytometry showed that 90% (P less than 0.01) of the analysed tumours with c-erbB-2 and c-erbA co-amplification, and 70% (P less than 0.1) of those with c-myc amplification were DNA aneuploid. This study demonstrates the potential value of c-myc amplification in the assessment of the tumour grade, rather than metastatic potential; and of the co-amplification of c erbB-2 and c-erbA as a strong indicator of metastatic potential, rather than tumour grade. PMID- 2572270 TI - High-level expression of MRK 16 and MRK 20 murine monoclonal antibody-define proteins (170,000-180,000 P-glycoprotein and 85,000 protein) in leukaemias and malignant lymphomas. AB - Using flow cytometry and immunocytochemistry, we investigated the reactivities of two different murine monoclonal antibodies (MAbs), MRK 16 and MRK 20, specific to adriamycin-resistant K562 cells (K562/ADM) with peripheral human mononuclear cells (MNC) (mainly blastic cells and lymphocytes) from 31 patients with leukaemia or malignant lymphoma. Reactivity with MRK 16 MAb was observed in five cases and reactivity with MRK 20 MAb in 18 cases. The cases were divided into three groups according to their reactivity patterns: group I, only the proportion of MRK 16-positive cells was increased; group II, only the proportion of MRK 20 positive cells was increased; group III, both MRK 16-and MRK 20-positive cells were increased. Some cases reflected the prior administration of adriamycin, vincristine, vinblastine and VP-16, which are known to induce P-glycoprotein expression. Expression of Mr 85,000 protein was observed more frequently than that of P-glycoprotein in leukaemia and malignant lymphoma, and this was not associated with either the total dose or period of administration of anticancer drugs. The expression of Mr 85,000 protein recognised by MRK 20 was further confirmed by Western blot analysis. PMID- 2572271 TI - Autografting for patients with chronic myeloid leukaemia in chronic phase: peripheral blood stem cells may have a finite capacity for maintaining haemopoiesis. AB - We treated 14 patients with Ph-chromosome-positive chronic myeloid leukaemia still in chronic phase by autografting with blood-derived haemopoietic stem cells. Eleven patients were autografted electively after cytoreductive treatment with busulphan (16 mg/kg) and melphalan (60 mg/m2) and three were autografted after marrow cells from HLA-identical sibling donors had failed to engraft. In 13 patients haemopoiesis recovered; one failed to engraft and died 114 d after autografting. Two other patients became pancytopenic and received further stem cell transfusions at 3 and 40 months respectively after first autografting. One patient entered lymphoid transformation and died 14 months after autografting. Twelve patients survive at a median of 41 months (range 24-53) after autografting. Nine of the survivors have required further chemotherapy after autografting and four of the nine were electively autografted on a second occasion. Three patients surviving after autografting for 28, 43 and 53 months respectively have not required further chemotherapy. In two of these patients haemopoiesis is now predominantly Ph-negative. We conclude that autografting in chronic phase might prolong survival in some cases by reducing the size of the leukaemic stem cell population. The fact that initially successful grafts failed in two patients suggests that blood-derived stem cells may have a finite potential for self-replication. PMID- 2572272 TI - Stimulation of glutathione synthesis in iron-loaded mice. AB - We have previously reported that the iron-loading of mice, by feeding them carbonyl iron, caused an elevation of hepatic glutathione concentration and an increase in glutathione excretion from the liver (Kawabata, T., Ogino, T. and Awai, M. (1989) Biochim. Biophys. Acta 1004, 89-94). To elucidate the mechanism of glutathione elevation, hepatic cysteine concentration and gamma glutamylcysteine synthetase (L-glutamate: L-cysteine gamma-ligase (ADP-forming), EC 6.3.2.2) activity were measured and possible changes in cysteine metabolism were also compared between iron-loaded and control mice. Hepatic cysteine concentration was higher in iron-loaded mice (185 +/- 12 nmol/g wet wt.) than in the controls (164 +/- 8 nmol/g wet wt.), and gamma-glutamylcysteine synthetase activity was also elevated in iron-loaded mice (34.3 +/- 3.2 nmol/mg protein per min) compared with the controls (28.6 +/- 3.8 nmol/mg protein per min). A comparison of the metabolic pathways with intravenously injected [35S]cysteine showed that organ distribution of the isotope was not significantly different, and also the rate of [35S]cysteine uptake into the hepatic glutathione fraction exhibited no difference between the two groups of mice. This shows that hepatic cysteine turnover may not be different between the two groups of mice. Since hepatic cysteine concentration was higher in iron-loaded mice, the apparently equal turnover of hepatic cysteine suggests that GSH synthesis may be elevated in iron-loaded mice. The high gamma-glutamylcysteine synthetase activity is suggested to stimulate GSH synthesis in iron-loaded mice. PMID- 2572274 TI - Electrokinetic, analytical and functional heterogeneity of circulating human platelets: separation of subpopulations by continuous flow electrophoresis after taxol stabilization. AB - A continuous flow electrophoresis procedure has been developed to study platelet subpopulation heterogeneity with separations based upon surface electrical charge differences. Taxol at low concentrations has been used to transiently stabilize the cells during the separations. At a concentration of 10(-5) M taxol has no effect upon a wide range of physical, analytical and enzymatic properties and does not compromise agonist-induced activation responses (aggregation and secretion). A typical normal platelet subpopulation profile extends over 15-20 fractions with mobilities from -0.97 to -0.78 microns per s per volt per cm. Platelet size (resistive particle counter volumes) differed significantly across the profile, the most electronegative cells being the larger, and the least electronegative the smaller platelets. Total platelet sialic acid content and surface neuraminidase-labile sialic acid correlated positively with electronegativity, but the surface -SH group status had an inverse relationship with the least electronegative smaller platelets, having twice as many surface DTNB-titratable - SH groups as the most electrophoretically mobile and larger cells. Normalisation of analytical and enzymatic data to cell volumes revealed that the smaller less electronegative platelets were substantially richer in all constituents and properties than the larger more electronegative platelets. These smaller cells showed higher activities for lysosomal enzymes, and their functions (capacity to transport 5-hydroxytryptamine and adenosine across the plasma membrane and responsiveness to thrombin expressed by synthesis of thromboxane B2 (TXB2) or release of 5HT) were greater than the larger more electronegative cells. No significant differences were observed, however, in the subpopulations by optical aggregometry using six different agonists each at three different concentrations. This free flow electrophoresis separation of platelets, which can be carried out on a preparative scale, may have some advantages over the conventional density gradient separations of subpopulations for investigating clinical states affecting thrombopoietic regulation or platelet losses from the circulation due to vessel wall disease, prosthetic implants or during extracorporeal circuitry. PMID- 2572273 TI - Influence of immunologic activation and cellular fatty acid levels on the catabolism of platelet-activating factor within the murine mast cell (PT-18). AB - The present study has examined the catabolism of 1-O-[3H]hexadecyl-2-acetyl-GPC (C16-PAF) and of 1-O-octadecyl-2-acetyl-GPC (C18-PAF) in spleen-derived PT-18 murine mast cells (mast cells). Mast cells catabolized exogenous PAF into two inactive metabolites, 1-O-alkyl-2-lyso-sn-glycero-3-phosphocholine (lysoPAF) and 1-O-alkyl-2-acyl-sn-glycero-3-phosphocholine (1-O-alkyl-2-acyl-GPC). The rate of conversion of C16-PAF to metabolites was more rapid than that of C18-PAF. Analysis of the acyl composition of 1-O-alkyl-2-acyl-GPC formed during the metabolism of PAF revealed that arachidonic acid (20:4) was the major fatty acyl chain incorporated at the sn-2 position. However, 25% of newly formed 1-O-alkyl-2 acyl-GPC was reacylated with docosahexaenoic acid (22:6). The influence of cellular fatty acid content on PAF catabolism was further explored in mast cells in which the ratio of fatty acids within cellular phosphoglycerides had been altered by supplementing the cells with various fatty acids in culture. Mast cells supplemented with 20:4 or 22:6 converted PAF to 1-O-alkyl-2-acyl-GPC at a significantly higher rate than non-supplemented cells. In contrast, cells supplemented with linoleic acid (18:2) metabolized PAF at rates similar to non supplemented cells. Analysis of the acyl composition of 1-O-alkyl-2-acyl-GPC derived from the metabolism of PAF in 20:4-supplemented cells indicated that 20:4 was incorporated exclusively into the sn-2 position. Conversely, 22:6 supplemented cells incorporated predominantly 22:6 at the sn-2 position of 1 alkyl-2-lyso-GPC. Supplementation with 18:2 had no effect on the acylation pattern seen in newly formed 1-O-alkyl-2-acyl-GPC. Activation of passively sensitized mast cells with antigen or with ionophore A23187 significantly enhanced the rate of catabolism of exogenously-provided PAF but had no effect on the acylation pattern of 1-O-alkyl-2-acyl-GPC. Experiments performed with the soluble fraction of the cells showed that acetyl hydrolase activity was increased in mast cells stimulated with antigen. In addition, supernatant fluids from antigen or ionophore-treated mast cells converted PAF to lysoPAF, suggesting that acetyl hydrolase activity was released during cell activation. These data indicate that the ability of mast cells to catabolize PAF to inactive metabolites is influenced by cell activation and by the cellular levels of certain fatty acids. PMID- 2572275 TI - In vitro activation of bovine adrenal tyrosine hydroxylase by rabbit skeletal muscle actin: evidence for a possible role of cytoskeletal elements as an activator for cytoplasmic enzymes. AB - Tyrosine hydroxylase prepared from the soluble fraction of bovine adrenal medulla was markedly activated by rabbit skeletal muscle G-actin, and this activation was accompanied by a decrease in the apparent Km of the enzyme for the pterin cofactor. The activating effect of G-actin on the soluble enzyme was still observed in the medium containing a high concentration of salt or excess amounts of proteinase inhibitors. Furthermore, this effect was not affected by either cytochalasin B or DNase I. These results therefore suggest that G-actin interacts with the enzyme molecule at the binding site(s) different from that involved in actin polymerization, and that it causes the activation of the soluble enzyme as a result of an allosteric alteration in the enzyme structure, thus giving rise to the possibility that cytoskeletal elements play an important role in the regulation of catecholamine synthesis as a factor modulating the activity of cytoplasmic tyrosine hydroxylase. PMID- 2572276 TI - Further characterization of the binding of plasminogen to heparin: evidence for the involvement of lysine residues. AB - We have previously demonstrated that the heparin-binding site of plasminogen is located in Val442-plasminogen region (kringle 5 domain plus light (B) chain) (Soeda, S., Kakiki, M., Shimeno, H., and Nagamatsu, A. (1987) Biochim. Biophys. Acta 916, 279-287). The chemical modification of Val442-plasminogen with a lysine reagent, pyridoxal 5'-phosphate (PLP), and sodium borohydride resulted in the incorporation of 8-10 PLP moieties per molecule of the zymogen. This PLP-labeled zymogen had no affinity for a heparin-Sepharose column, whereas the non-labeled one bound to the column. Modification in the presence of heparin decreased the extent of labeling by 1-2 mol of PLP per mol of Val442-plasminogen. To further examine the binding site of plasminogen to heparin, functionally active A and B chains were separated from Lys-plasmin after mild reduction and S carboxymethylation. Only B chain possessed affinity for heparin-Sepharose. Furthermore, plasmin(ogen) bound to heparin was protected from alpha 2 antiplasmin inhibition. These results indicate that one or two lysine residues located in the catalytic region (B chain) of plasmin(ogen) are essential to heparin binding, and that the binding of plasminogen to heparin or heparin-like substance in extracellular matrix environments may be important for the localization and activation of plasminogen and for the prolongation of the resultant plasmin activity. PMID- 2572277 TI - An improved procedure for the purification of formiminotransferase-cyclodeaminase from pig liver. Kinetics of the transferase activity with tetrahydropteroylpolyglutamates. AB - Formiminotransferase-cyclodeaminase is stabilized and activated approx. 40% in the presence of low concentrations (equal or less than 0.2%) of Triton X-100, possibly because the average hydrophobicity (1.10 kcal per residue) and the frequency of large non-polar side-chains (0.34) of this protein are both somewhat higher than average. This stabilization enabled us to develop a new purification procedure for the enzyme using chromatography on Matrex Gel Orange A and heparin Sepharose columns in the presence of Triton X-100. This procedure is easier, much more reproducible, and gives slightly higher yield than the previous method described by Drury, et al. Further investigations of the role of tetrahydropteroylpolyglutamates with formiminotransferase-cyclodeaminase reveal that the use of polyglutamylated folate substrates does not change the mechanism of the transferase reaction, but decreases the K(m) for formininoglutamate, the second substrate, more than 10-fold, bringing it closer to the expected physiological concentration. PMID- 2572278 TI - Perinatal activity of hepatic gamma-glutamyltranspeptidase in rats from large and small litters. AB - In rat pups, the activity of hepatic gamma-glutamyltranspeptidase (GGT) is not influenced by the process of labor. Maximum activities are observed prenatally occurring at the 21st day of gestation in fetuses from normal, large litters, but at the 20th day of gestation in fetuses from experimentally reduced litters (2 fetuses per litter). This different developmental pattern may depend on processes of fetal maturation rather than on processes of luteolysis. PMID- 2572280 TI - Changes of subcellular localization of Thy-1 antigen during thymic myoid cell differentiation. AB - Using a quantitative enzyme immunoassay, Thy-1 antigen expressed by a rat myoid cell line R615B2 was detected mainly on the cell surface at a single cell stage, whereas at the stage of forming myotubes, Thy-1 was found predominantly in the cytoplasm. The muscle specific creatine kinase activity also increased in association with the shift of Thy-1 from the cell surface to the cytoplasm, suggesting biological significance of Thy-1 redistribution in muscle differentiation from single cells to multinucleated cells. PMID- 2572279 TI - Adrenergic stimulation of inositol-phosphate production in a genital tract smooth muscle cell line. AB - Catecholamines are important in the modulation of smooth muscle contractile activity; this study was undertaken to evaluate adrenoceptor stimulation of intracellular inositol-phosphate production in a genital tract smooth muscle myocyte. DDT1 MF-2 smooth muscle myocytes, derived from a hamster ductus deferens leiomyosarcoma, were loaded with 3H-inositol, incubated in 10 mM LiCl, then stimulated with adrenergic agonists with and without antagonists. Subsequently, the inositol phosphates were isolated by anion-exchange chromatography. In the presence of norepinephrine (NE), inositol trisphosphate (IP3) was produced by 30 s and peaked at 2 min; inositol 1-phosphate was also apparent by 30 s, and continued to increase over 15 min. Clonidine (an alpha-2 agonist), isoproterenol, and NE in the presence of phentolamine or prazosin (an alpha-1 antagonist) failed to increase IP3. In contrast, NE in the presence of yohimbine (an alpha-2 antagonist) or propranolol stimulated IP3 production to levels comparable to that stimulated by NE alone. These studies provide evidence that inositol phosphate production is involved in alpha-1 adrenergic signal transduction in DDT1 MF-2 myocyte. PMID- 2572281 TI - [Dipeptidyl aminopeptidase-IV in lymphocytes of patients with lymphoproliferative diseases]. AB - DAP-IV activity (Gly-Pro-MCA hydrolysis, pH 7.8) was found in lysates of peripheral blood lymphocytes of patients with T- and B-cell forms of malignant lymphoproliferative diseases. The highest DAP-IV activity was seen in the cells of patients with a rare variant of T-cell lymphocytic leukemia (T-CLL); these cells expressed simultaneously the antigens of T helpers and T suppressors (Th and Ts) (OKT4+ and OKT8+). The DAP-IV activity about ten times less was found in the pathological cells with a phenotype of mature Th (Sezary disease), as well as in the cells expressing antigens of both Ts and natural killers (a rare variant of T-CLL). The same activity was also found in Ts (T gamma-lymphocytosis). The data obtained show that the differences in DAP-IV expression are connected with the differentiation step rather than with the belonging to a particular subpopulation of T-cells. DAP-IV activity, which was somewhat lower than that of T-cells, was found in B-lymphocytes of patients with B-CLL, hair-cellular leukemia, and non-Hodgkin's lymphoma. No correlation of DAP-IV activity with the level of E-cellular differentiation was observed. PMID- 2572282 TI - 2'-Deoxycytidine protects normal human bone marrow progenitor cells in vitro against the cytotoxicity of 3'-azido-3'-deoxythymidine with preservation of antiretroviral activity. AB - Bone marrow cytotoxicity of 3'-azido-3'-deoxythymidine (AZT), an anti-human immunodeficiency virus (anti-HIV) drug, has been attributed to deoxyribonucleotide pool perturbations that might result in impaired DNA synthesis in normal bone marrow elements. We examined, in vitro, the effect of high, but clinically achievable and nontoxic, concentrations of 2'-deoxycytidine (dCyd) (greater than or equal to 100 mumol/L) on high-dose AZT mediated growth inhibition and intracellular biochemical perturbations in normal bone marrow progenitor cells. Colony formation by bone marrow progenitor cells in semisolid medium was significantly protected by dCyd against the inhibitory effects of co administered, high concentrations of AZT (10 mumol/L). Also, dCyd significantly corrected AZT mediated depletion of intracellular thymidine triphosphate (dTTP) and dCyd triphosphate (dCTP) levels in normal bone marrow mononuclear cells (BMMC). Moreover, dCyd reduced the intracellular accumulation of AZT triphosphate (AZT-TP) and its DNA incorporation in BMMC. In contrast, co-administration of dCyd (100 mumol/L to 1 mmol/L) did not reverse AZT (10 mumol/L) mediated suppression of HIV infectivity in HUT-102 cells in culture, although a partial reduction in intracellular AZT-TP pools and its DNA incorporation as well as a correction of AZT mediated depletion of dTTP and dCTP pools was observed in these cells. These studies suggest that dCyd at high concentrations might ameliorate the bone marrow cytotoxicity of high-dose AZT without impairing its anti-HIV effect. PMID- 2572283 TI - Elimination of drug-resistant myeloma tumor cell lines by monoclonal anti-P glycoprotein antibody and rabbit complement. AB - The effectiveness of ex vivo chemotherapy with drugs, such as vincristine, etoposide, and Adriamycin (doxorubicin, Adria Labs, Columbus, OH) for elimination of residual tumor cells from human bone marrow grafts could be undermined by the presence of multidrug-resistant tumor cells in the bone marrow. Therefore, to supplement chemoseparation, we investigated whether MRK-16, a monoclonal antibody (MoAb) to the surface moiety of multidrug resistance-associated P-glycoprotein antigen, can eliminate drug-resistant tumor cells in the presence of rabbit complement (RC). Two doxorubicin (DOX)-resistant human myeloma tumor cell line, 8226/DOX40 (resistant to 4 x 10(-7) mol/L DOX) and 8226/DOX6 (6 x 10(-8) mol/L DOX) with high and low amounts of cell surface P-glycoprotein, respectively, and the drug-sensitive parent cell line 8226/S were used as tumor models in this study. Using the limiting dilution assay, we have shown that three cycles of treatment with 25 micrograms/mL of MRK-16 MoAb and a 1:4 final dilution of RC eliminated 2.90 +/- 0.10 logs of 8226/DOX40 cells and 1.94 +/- 0.18 logs of 8226/DOX6 cells. One and two cycles of treatment were less effective, eliminating 0.47 +/- 0.40 and 1.94 +/- 0.36 logs of 8226/DOX40 and 0.12 +/- 0.20 and 1.63 +/- 0.58 logs of 8226/DOX6 cells, respectively. The 8226/S cell growth was unaffected by one to three cycles of treatment. The cell kill was not impaired when the antibody plus complement treatment was carried out on a mixture of 8226/DOX40 or 8226/DOX6 cells with a ninefold excess of irradiated bone marrow mononuclear cells (MNCs). The three cycles of treatment with antibody plus complement did not adversely affect granulocyte-macrophage colony-forming unit (GM-CFU) survival in hematologically normal marrows (92.5% to 104% survival) or in myeloma patient marrows (85% to 100%). These results show that it is possible to eliminate drug resistant myeloma tumor cell lines from the admixed human bone marrow by treatment with MRK-16 MoAb plus RC. This method could prove to be effective for elimination of other drug-resistant tumor cell lines including those of leukemia and solid tumors, and will be further useful for supplementing chemopurging, and immunopurging of bone marrow with other antitumor cell antibodies. PMID- 2572284 TI - T-cell ontogeny after autologous bone marrow transplantation: failure to synthesize interleukin-2 (IL-2) and lack of CD2- and CD3-mediated proliferation by both CD4- and CD8+ cells even in the presence of exogeneous IL-2. AB - T cells generated during a second round of ontogeny after autologous bone marrow transplantation (ABMT) represent a unique model of early T-cell ontogeny in an autologous situation. Since grafted bone marrows were pretreated in vitro with the cyclophosphamide derivative ASTA Z 7557, circulating T cells had to be regenerated from reinfused hematopoietic progenitor cells. The T-cell population derived from 25 patients post-ABMT was phenotypically characterized: an increase in CD8+ cells, a low percentage of CD4+ cells, and a median of 12% CD56+ (NKH1+) cells were found. When the T cells were stimulated with phytohemagglutinin (PHA) and phorbol myristate acetate (PMA), defective interleukin-2 (IL-2) secretion was observed. In addition, proliferative responses of the T cells after activation through the antigen-receptor-dependent CD3 pathway, through the CD2 dependent alternative T-cell pathway, and by the lectin PHA were investigated. Despite the presence of CD2, CD3, alpha/beta chains of the T-cell receptor, and CD25+ IL-2 surface receptors, abnormal proliferative responses were obtained even in the presence of exogeneous IL-2. In experiments where the T-cell population was separated into CD4+ cells and CD8+ cells, both the CD4- and CD8+ subsets were unable to respond to activating and proliferating signals. Thus, T cells at early stages of ontogeny not only possess an intrinsic defect in IL-2 synthesis but, in addition, were unable to express functional IL-2 receptors in response to mitogenic stimuli. PMID- 2572285 TI - Third-party-mediated graft rejection and graft-versus-host disease after T-cell depleted bone marrow transplantation, as demonstrated by hypervariable DNA probes and HLA-DR polymorphism. AB - Graft rejection after marrow transplantation is generally thought to be mediated by alloreactive immune effector cells of host origin. Transfused blood products also contain immune cells capable of alloreactivity against both donor graft and host. To reduce the risk of transfusion-associated graft-versus-host disease (GVHD) and graft rejection, standard procedure is to irradiate all blood products with at least 1,500 rad before transfusion. We report a patient with chronic myelogenous leukemia who developed graft rejection and GVHD after receiving a T cell-depleted transplant from a serologically HLA-A, B, DR/DQ matched and mixed lymphocyte culture (MLC) nonreactive unrelated donor. Cytogenetic analysis of marrow cells collected at the time of graft rejection revealed a PH1-negative female karyotype that was not consistent with donor cells. Use of specific minisatellite DNA probes (YNH 24, H-RAS, and 3' HVR) revealed the exclusive presence of third-party (neither donor nor recipient) restriction-fragment-length polymorphisms (RFLP) in both peripheral blood and marrow. Repeat RFLP analysis 3 days later showed persistence of this unique third-party banding pattern. DNA based HLA-typing, using polymerase chain reaction (PCR) and oligonucleotide probe hybridization, also showed these cells to be derived from an individual whose HLA DR type was distinct from donor and recipient. Together, these findings suggested the presence of a proliferating population of transfused cells possessing alloreactivity against both donor graft and host, despite prior irradiation of all blood products with 2,000 rad. Limiting dilution analysis to assess the frequency of irradiated lymphocytes able to respond to mitogen revealed an approximate 5- to 6-log reduction at 1,500 to 2,000 rad as compared with unirradiated controls. These data indicate that a small percentage of lymphocytes can survive irradiation at these doses and suggest that existing blood-product irradiation guidelines may require reassessment, especially in T-cell-depleted transplant recipients. PMID- 2572286 TI - Rearrangements in the p53 gene in Philadelphia chromosome positive chronic myelogenous leukemia. AB - Molecular structural analysis of the p53 gene in patients with Philadelphia chromosome-positive chronic myelogenous leukemia (CML) indicates a significant incidence of gene rearrangements in patients at either accelerated phase or blastic crisis. Southern blot analysis of genomic DNA hybridizing with either genomic or cDNA p53 specific probes indicated that 30% of the CML patients at blastic crisis phase exhibited rearrangements, mostly mapping downstream to the first non-coding exon. This is compatible with the observation that the progression of CML from the chronic to the acute phase involves frequent aberrations in chromosome 17, to which the p53 oncogene has been mapped. Therefore, we suggest that one of the pathways of development of CML to the acute phase is associated with aberrations in the p53 nuclear oncogene. PMID- 2572287 TI - Cell-type-specific cytotoxicity of anti-CD4 and anti-CD8 ricin immunotoxins against human alloreactive T-cell clones. AB - Potent T-cell subset-directed immunotoxins (ITs) were generated by conjugating the anti-CD4 monoclonal antibody (MoAb) G17-2 and the anti-CD8 MoAb G10.1 to the ribosome-inhibitory protein, ricin. The cell-type-specific cytotoxicities of the generated ITs were evaluated at the clonal level using human alloreactive T-cell clones. The kinetics of anti-CD4 ricin-induced inactivation of protein synthesis in target CD4+ cloned T-cells was first order with no detectable lag period and a maximum rate of 0.07 logs per hour (t10 = 13.6 hours; first-order rate constant/K = 0.17 hr-1). The alloantigen specific lytic function of the CD4+ cytolytic T cell clone JMAC28 was acutely sensitive to anti-CD4 ricin, and no residual lytic activity against allogeneic targets was detectable 24 hours after treatment with as little as 0.5 mmol/L anti-CD4 ricin. Notably, both anti-CD4 ricin and anti-CD8 ricin elicited a selective and dose-dependent inhibition of clonal proliferation of target T-cell clones with a maximum kill of greater than 3 logs at 5 nmol/L. No significant "bystander effects" were observed for non-target cells. Bone marrow progenitor cells CFU-GM, BFU-E, and CFU-GEMM were only minimally affected by either IT. We conclude that these ITs show considerable potential for effective depletion of T-cell subpopulations from allogeneic donor marrow grafts for clinical graft-versus-host disease (GVHD) prophylaxis. PMID- 2572289 TI - Mumps, measles, and rubella vaccination and encephalitis. PMID- 2572288 TI - Molecular heterogeneity of glucose-6-phosphate dehydrogenase A-. AB - Glucose-6-phosphate dehydrogenase (G6PD) deficiency is probably the most common disease-producing genetic polymorphism of humans. Virtually all G6PD-deficient Africans show the G6PD A- phenotype, an electrophoretically rapid, deficient enzyme. The recently acquired ability to identify the point mutations producing the different variants has given us new insights into the population genetics of G6PD variants. Twenty-nine males with the G6PD A- phenotype were studied. They were of African, Mexican, Spanish, and US white ethnic origin. All had the A---G transition at nucleotide 376 characteristic of G6PD A. In each case, one of three additional mutations was present, at nucleotides 202, 680, or 968. That in this population second mutations producing G6PD deficiency occurred only on the genetic background of G6PD A suggests that G6PD A was at one time the most common type of G6PD in Africa. However, the nucleotide sequence of the chimpanzee (Pan troglodytes) G6PD indicates that the primordial human type of G6PD was G6PD B. PMID- 2572290 TI - Neutral aminopeptidase: a potential marker enzyme of the amphibian urinary bladder epithelial cell apical membrane. AB - Antidiuretic hormone induces, in the apical plasma membrane of amphibian urinary bladder epithelial cells, the exocytotic insertion of intramembranous particle aggregates that probably contain water channels. Purification of the apical membrane is a way to characterize the aggregates. The isolation of such purified membranous fractions involves the use of specific exogenous or endogenous markers. One of them could be the neutral aminopeptidase (AP), whose activity was detected in urinary bladder. Enrichment in AP activity was observed in plasma membrane preparations compared to cell homogenates (X2.7). However, a large part of the enzyme activity was also recovered in the soluble fraction of the preparation, suggesting large proteolysis of the protein. The enzyme presents a low optimal pH (6.4) and a high specificity for proline-p-nitroanilide as compared to the AP present in kidneys and intestines. To localize the protein in the amphibian bladder epithelium, an immunological approach was necessary due to the low activity of the enzyme in this tissue. The low enzymatic activity also prevented the purification of sufficient amounts of the urinary bladder AP as antigen, and we prepared antibodies against purified AP from frog or toad kidneys where the activity is 60 times higher than in the bladder. The serum specificity was verified by spot immunodetection, Western blot, inhibition capacity of antibodies, and immunoadsorption on a solid support with the renal enzyme. The sera were found to be able to react with native as well as denatured forms of the kidney enzyme. Antibodies cross-reacted with several peptides of low molecular weight (40-60 kDa) from urinary bladder plasma membrane proteins (Western blot).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572291 TI - Presynaptic beta-adrenoceptors in rat atria: evidence for the presence of stereoselective beta 1-adrenoceptors. AB - 1. Presynaptic beta-adrenoceptor activity was studied in rat isolated atria, previously loaded with [3H]-noradrenaline. The stimulation-induced release of 3H transmitter was measured in the presence of cocaine, and adrenaline was used as a facilitatory beta-adrenoceptor agonist. 2. Adrenaline (0.1 and 2 nM) increased, by about 50%, the evoked efflux of tritium. With phenoxybenzamine present, the same activity was shown with 10 nM adrenaline. 3. The beta 2-selective adrenoceptor blocking drugs: IPS 339 and ICI 118 551 caused a concentration dependent decrease in the activity of adrenaline. Cardioselective beta-blocking drugs: acebutolol, beta-xolol, nebivolol and its isomers (R 67 138 and R 67 145) also reduced dose-dependently the agonistic action of adrenaline. The order of potency for nebivolol and its isomers was R 67 138 greater than nebivolol greater than R 67 145. The activity of pindolol was not concentration-dependent. The inhibitory effect of acebutolol was also observed in the presence of blockade of alpha-adrenoceptors. 4. The postsynaptic beta-adrenoceptor blocking activity of nebivolol and its isomers was studied in pithed rats. They reduced isoprenaline induced tachycardia without altering hypotensive responses. The order of potency was: R 67 138 greater than nebivolol greater than R 67 145. 5. It is concluded that in rat isolated atria, presynaptic beta 2- and beta 1-adrenoceptors coexist and that facilitatory beta 1-adrenoceptors are stereospecific. PMID- 2572292 TI - Mechanisms underlying the antiarrhythmic properties of beta-adrenoceptor blockade against ischaemia-induced arrhythmias in acutely prepared rats. AB - 1. The mechanism underlying the limited antiarrhythmic effects of beta adrenoceptor blocking agents against occlusion-induced arrhythmias in acutely prepared, pentobarbitone-anaesthetized rats has been investigated. 2. Atenolol, ICI 111,581 and propranolol were given at low, medium and high doses calculated to shift dose-response curves to exogenous agonists by factors of 10-30, 100-300 and 1000-3000, respectively. 3. Arrhythmias, blood pressure, heart rate, ECG changes and serum K+ were measured. 4. Antiarrhythmic activity was seen with beta blocker treatment. This was minimal with atenolol (0.1, 1 and 10 mg kg-1) and only statistically significant with the highest dose of ICI 111,581 (5 mg kg-1), and propranolol (10 mg kg-1). 5. Treatment with beta-adrenoceptor blockers elevated serum potassium concentrations, as compared with saline controls, especially when measured at 30 min post-occlusion. 6. Only ICI 111,581 (5 mg kg 1) and propranolol (1 and 10 mg kg-1) prolonged P-R interval. 7. In order to evaluate possible mechanisms of antiarrhythmic action, attempts were made to correlate antiarrhythmic activity with beta-blockade, serum potassium concentrations, and/or with changes in the P-R interval of the ECG. 8. Reductions in arrhythmias did not correlate well with presumed beta-blockade. Better correlation was obtained with elevations of serum potassium concentration, and with prolongation of P-R interval (a presumed Class I antiarrhythmic action). 9. These results suggested that antiarrhythmic effects of adrenoceptor blocking agents in acutely-prepared anaesthetized rats, subjected to occlusion of a coronary artery, are unrelated to cardiac beta-blockade. The limited antiarrhythmic effects which were observed could be attributed to elevations in serum potassium concentration (due to peripheral beta-blockade) and/or possible Class I antiarrhythmic actions. PMID- 2572293 TI - Changes of the membrane potential in striatal synaptoneurosome, synaptosome and membrane sac preparations induced by glutamate, kainate and aspartate as measured with a cyanine dye DiS-C2-(5). AB - The effects of glutamate, kainate and aspartate on the membrane potential of striatal synaptoneurosome, synaptosome and membrane sac preparations were studied by using a potential sensitive cyanine dye DiS-C2-(5). Excitatory amino acids glutamate and aspartate had a depolarizing effect on synaptoneurosomes. 7.9 microM glutamate and 2.8 microM aspartate produced a half-maximal response. Depolarizations induced by glutamate and aspartate were dependent on the concentration of extracellular sodium ions, a maximal response occurred at around 40 mM of external Na+. Kainate induced a dual effect on synaptoneurosomes. In a standard Na+-based medium a hyperpolarization, likely due to inhibition of a presynaptic sodium-dependent glutamate uptake, predominated over a postsynaptic kainate receptor-mediated depolarization that was observed when electrogenic glutamate uptake was inhibited. This interpretation was supported by results obtained with synaptosome and membrane sac preparations. In a standard Na+-based medium kainate had a hyperpolarizing effect on synaptosomes while in the membrane sac preparation kainate induced a depolarization. PMID- 2572294 TI - Long-term treatment with antipsychotics does not alter the phosphoinositide response to muscarinic or D2 dopaminergic agonists in rat striatum. AB - The effects of the muscarinic agonist carbachol and the D2 dopaminergic agonist quinpirole on phosphoinositide hydrolysis were studied in the corpus striatum of rats which had been treated for one year with either haloperidol or clozapine. In the presence of LiCl, carbachol increased the accumulation of inositol monophosphate (greater than 100%) and bisphosphate (greater than 20%). Quinpirole had no effect on either basal or carbachol-stimulated accumulation of inositol phosphates. There was no difference in these responses between the drug-treated animals and age-matched controls. PMID- 2572296 TI - Frontal decortication decreases the affinity of N-(1-[2 thienyl]cyclohexyl)[3H]piperidine binding to rat striatum. AB - Bilateral ablation or transection of the corticostriatal pathways of the rat caused a reduction in N-(1-[2-thienyl]cyclohexyl)[3H]piperidine ([3H]TCP) binding to the striatum at 5-7, but not 3 and 14-28, days postlesion with a persistent decrease in striatal glutamate content 3-28 days after the decortication. This reduction was found to be due to an increase in Kd without changes in Bmax of the [3H]TCP binding. The present findings suggest that interruption of striatal glutamatergic transmission following frontal decortication may produce a temporal reduction in the affinity of phencyclidine receptor in the striatum. PMID- 2572295 TI - Somatostatin-like immunoreactivity in rat main olfactory bulb: extent of coexistence with neuropeptide Y-, tyrosine hydroxylase- and vitamin D-dependent calcium binding protein-like immunoreactivities. AB - A double-labeling immunofluorescence colocalization technique was used to examine the extent of coexistence of somatostatin (SOM)-like immunoreactivity with neuropeptide Y (NPY)-, tyrosine hydroxylase (TH)- and vitamin D-dependent calcium binding protein (D-CaBP)-like immunoreactivities in neurons of the rat main olfactory bulb. SOM-like immunoreactivity (SOM-I) was distributed within restricted populations of periglomerular neurons and deep short-axon cells, and rarely within superficial short-axon cells at the glomerular layer/external plexiform layer (GL/EPL) border region. Double-labeling analysis revealed that all of the SOM-I deep and superficial short-axon cells also contained NPY-I. Colocalization of SOM-I and TH-I or of SOM-I and D-CaBP-I was infrequently observed within periglomerular neurons. The rare SOM-I short-axon cells at the GL/EPL border always exhibited D-CaBP-I. These results demonstrate virtual complete coexistence of SOM and NPY in short-axon neurons of the main olfactory bulb. With a few exceptions, however, bulbar SOM neurons appear to constitute subclasses of periglomerular cells immunohistochemically distinct from those containing TH or D-CaBP. PMID- 2572297 TI - Electrophysiological characterization of adrenoceptors in the rat dorsal hippocampus. III. Evidence for the physiological role of terminal alpha 2 adrenergic autoreceptors. AB - The present electrophysiological studies were undertaken to assess the role of terminal alpha 2-adrenergic autoreceptors in regulating noradrenergic synaptic transmission in the rat CNS. The effectiveness of the electrical stimulation of the locus coeruleus (LC) in suppressing the firing activity of pyramidal neurons was determined in the dorsal hippocampus. Intravenous clonidine, an alpha 2 adrenergic agonist, decreased the effectiveness of the LC stimulation, without altering the effect of microiontophoretically applied norepinephrine. The subsequent i.v. administration of low doses of idazoxan, an alpha 2-adrenergic antagonist, reversed this effect of clonidine on the LC stimulation. To ascertain that the effect of clonidine administered i.v. was indeed attributable to its action on noradrenergic terminals, it was applied locally by microiontophoresis; it decreased the effectiveness of the LC stimulation. Another paradigm used to assess the function of terminal alpha 2-adrenoceptors was to increase the frequency of the LC stimulation from 1 to 5 Hz. This resulted in a 5-fold decrease of the effectiveness of the stimulation. That this was attributable to an enhanced activation of terminal alpha 2-adrenoceptors was suggested by the reversal of this effect of increasing the frequency of the LC stimulation by intravenous idazoxan. Furthermore, the degree of enhancement of the effectiveness of the LC stimulation by idazoxan was much greater at 5 than at 1 Hz. These results provide novel electrophysiological evidence for the potent regulatory role of terminal alpha 2-adrenoceptors on noradrenergic neurotransmission. PMID- 2572299 TI - Excitatory amino acid neurotoxicity at the N-methyl-D-aspartate receptor in cultured neurons: role of the voltage-dependent magnesium block. AB - Results of the present report show that cerebellar neurons in primary culture are resistant to glutamate concentrations as high as 5 mM in the presence of glucose and Mg2+, but sensitive to glutamate concentrations lower than 35 microM when the neurons are deprived of glucose. Glutamate toxicity is also potentiated when Mg2+ is removed but glucose and EDTA are present; in this case, higher concentrations of glutamate (1 mM) are required for full toxicity. Glucose concentrations as low as 50 microM are fully protective against the toxicity of 100 microM glutamate; pyruvate and, to a lesser extent, lactate are also protective. Significantly, increasing concentrations of extracellular Mg2+ are fully protective against the toxicity of 100 microM glutamate in the absence of glucose and against the toxicity of 1 mM glutamate in the presence of glucose and EDTA. We interpret these results as support for our hypothesis that the pivotal event in glutamate's transition to neurotoxin is relief of the Mg2+ block of the N-methyl-D-aspartate (NMDA) receptor channel, which is known to be voltage-dependent. Partial depolarization in response to depletion of high-energy phosphates relieves the voltage-dependent block enabling glutamate to stimulate an excessive ion influx which results in the death of the neuron by a mechanism which is not yet understood. We propose that this mechanism may be operative in the neuronal damage associated with a variety of neurodegenerative disorders. PMID- 2572298 TI - Excitatory amino acid neurotoxicity at the N-methyl-D-aspartate receptor in cultured neurons: pharmacological characterization. AB - L-Glutamate neurotoxicity at the N-methyl-D-aspartate (NMDA) receptor was characterized in cultured cerebellar granule cells. When deprived of glucose for 40 min, these cells were killed by 20-60 microM L-glutamate. However, the neurons were resistant to glutamate at concentrations as high as 5 mM when glucose and Mg2+ were present throughout. Both competitive and non-competitive NMDA receptor antagonists completely blocked neurotoxicity due to glutamate and other NMDA receptor agonists. CPP [+/-)-3-(2-carboxypiperazin-4-yl)-prophyl-1-phosphonic acid) was the most effective competitive antagonist with full protection at 100 microM while MK-801 [+/-)-10,11-dihydro-5-methyl-5H-dibenzo[a,d]-cyclohepten-5,10 imin e) was the most effective non-competitive antagonist with full protection at 20 nM. Other antagonists with higher selectivity for other subtypes of glutamate receptors were ineffective. We conclude that glutamate toxicity in energy deprived cerebellar granule cells is mediated by NMDA receptors. Results are discussed in terms of an hypothesis offering an explanation for the transition of glutamate from neurotransmitter to neurotoxin which emphasizes the responsiveness of the receptor to agonists rather than focusing on the presence of high concentrations of agonist. PMID- 2572300 TI - In vivo tyrosine hydroxylation rate in retina: effects of phenylalanine and tyrosine administration in rats pretreated with p-chlorophenylalanine. AB - p-Chlorophenylalanine was administered to rats to inhibit hepatic phenylalanine hydroxylase activity. Two days later, phenylalanine injection was noted to produce substantial increases in serum phenylalanine levels, and relatively modest increments in serum tyrosine levels. Rats injected with p chlorophenylalanine 2 days earlier showed a normal light-induced activation of retinal tyrosine hydroxylase activity in vivo, measured as dihydroxyphenylalanine accumulation following pharmacologic inhibition in vivo of aromatic L-amino acid decarboxylase activity. In addition, tyrosine injection into p chlorophenylalanine-treated rats in the light produced anticipated increments in retinal tyrosine hydroxylation rate, showing the enzyme to be functionally normal. The acute administration of phenylalanine (62.5-500 mg/kg i.p.) to p chlorophenylalanine-treated rats produced dose-related increments in retinal phenylalanine. In vivo tyrosine hydroxylation rate in retina was normal at all doses below 300 mg/kg. However, at the highest dose (500 mg/kg), when retinal phenylalanine levels were almost 5-times normal tyrosine hydroxylation rate consistently fell (to about half-normal values). These results demonstrate that very large elevations in tissue phenylalanine levels do not stimulate tyrosine hydroxylation in vivo, and that at extremely high levels phenylalanine inhibits tyrosine hydroxylation rate. PMID- 2572301 TI - Dynorphin A(1-13) modulates apomorphine-induced behaviors using multidimensional behavioral analyses in the mouse. AB - The effects of intracerebroventricular injection of dynorphin A(1-13) on apomorphine-induced behavioral changes were investigated in the mouse using multidimensional behavioral analyses based upon a capacitance system. Although lower doses (0.1 or 0.3 mg/kg) of apomorphine were without marked effects on behaviors, a 0.56 mg/kg dose of the drug evoked a significant increase in rearing behaviors. Furthermore 1.0 and 3.0 mg/kg doses of apomorphine produced a marked increment in linear locomotion, circling and rearing. Dynorphin A(1-13) (3.0 or 10.0 microgram) itself had no effects on behaviors. The apomorphine (0.56 and 1.0 mg/kg)-induced increase in rearing behaviors was clearly inhibited by treatment with dynorphin A(1-13) (3.0 and 10.0 microgram). Simultaneously, the marked increases in linear locomotion and circling were displayed by apomorphine (1.0 mg/kg) plus dynorphin A(1-13) (10.0 microgram). The effects of dynorphin A(1-13) (10.0 microgram) on the apomorphine (1.0 mg/kg)-induced increase in rearing were entirely reversed by the opioid antagonist Mr2266. These results suggest that the antagonistic effects of dynorphin A(1-13) on the apomorphine (1.0 mg/kg)-induced increase in rearing are mediated via opioid receptors, possibly K-sites. PMID- 2572303 TI - Ethanol inhibits NMDA-induced increases in free intracellular Ca2+ in dissociated brain cells. AB - The effect of N-methyl-D-aspartate (NMDA) on free intracellular Ca2+ concentrations [( Ca2+]i) and the interaction of ethanol on the NMDA-mediated response was examined in freshly dissociated brain cells isolated from newborn rats. NMDA (25 microM) increased [Ca2+]i by approximately 70 nM, measured by fura 2 fluorometry, and this increase could be prevented or reversed by the NMDA antagonists Mg2+ (1.0 mM) and 2-amino-5-phosphonovalerate (AP5, 100 microM). Ethanol (25, 50, 100 mM) added 50 s before NMDA (25 microM) reduced the rise in [Ca2+]i when compared to the 25 microM NMDA response in the absence of ethanol. Thus, ethanol may have direct actions on NMDA-receptor activated increases in [Ca2+]i. PMID- 2572302 TI - Labeling of NMDA receptor channels by [3H]MK-801 in brain synaptic membranes treated with Triton X-100. AB - Binding activity of [3H](+)-5-methyl-10, 11-dihydro-5H-dibenzo [a,d] cyclohepten 5, 10-imine maleate (MK-801) was examined by using rat brain synaptic membranes treated with Triton X-100. This compound is known as a non-competitive antagonist for one subclass of the central excitatory amino acid receptors, N-methyl-D aspartic acid (NMDA)-sensitive receptors. Triton treatment completely abolished the temperature-dependent portion of the binding activity, with a concomitant reduction of membranous protein content. Addition of L-glutamic acid (Glu), however, markedly potentiated the activity at concentrations higher than 10 nM in a temperature-dependent manner. Similarly significant potentiation was induced by structurally related amino acids as well as agonists for the NMDA-sensitive receptors, but not by agonists for the other subclasses. The rank order of this stimulatory potency was well consistent with that of the displacing activity of these Glu analogues on NMDA-sensitive [3H]Glu binding. Competitive NMDA antagonists, such as (+/-)-3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid and D-2-amino-5-phosphonovaleric acid, were all effective in preventing the potentiation of [3H]MK-801 binding activity by L-Glu. The latter Glu-dependent activity was additionally enhanced by glycine and its analogues at concentrations above 10 nM in a temperature- and Glu-dependent fashion. Additional enhancement by glycine was also antagonized by competitive NMDA antagonists, but not by a classical glycine antagonist strychnine. These results suggest that Triton X-100 treated membranes are suitable for the study of the interaction of the non competitive antagonists with NMDA receptor channels, and may be superior to non detergent-treated membranes in terms of freedom from the confounding effects of endogenous amino acids. PMID- 2572304 TI - Neuropathological changes in the caudate nucleus elicited by MPTP and their prevention by monoamine oxidase inhibition. AB - MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine), a neurotoxin producing parkinsonism, causes an obvious reduction in tyrosine hydroxylase (TH) activity in the caudate nucleus. This depletion of TH activity is due to degeneration of TH-positive punctate dopaminergic terminals. The disappearance of these terminals unmasks the presence of long branching, varicose preterminal fibers, which may be sprouting after degeneration of their terminal arborizations. Glial cells, normally sparse with delicate processes, undergo intense increase in number and a robust hypertrophy. All of these changes are prevented completely by administration of deprenyl, a monoamine oxidase inhibitor, before and after MPTP. These neuropathological effects are additional manifestations of the dopaminergic neurotoxicity induced by MPTP. The metabolism of MPTP, which is blocked by monoamine oxidase inhibition, is apparently necessary for the expression of toxicity in the brain by this neurotoxin. PMID- 2572305 TI - MPTP treatment combined with ethanol or acetaldehyde selectively destroys dopaminergic neurons in mouse substantia nigra. AB - We have previously reported that ethanol and acetaldehyde potentiate 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP) neurotoxicity in mice, enhancing dopamine (DA) depletion in the striatum. The present study was designed to determine whether such enhancement of neurotoxicity was specific for the nigro striatal DA pathway. In 5-week-old mice acetaldehyde treatment did not enhance DA depletion seen 7 days after MPTP treatment. In 8-week-old animals, however, acetaldehyde or ethanol given with MPTP decreased striatal DA content to about 10% of controls, whereas the depletion was to 43% of controls when MPTP was given alone. In acetaldehyde or ethanol and MPTP-treated mice, changes in DA levels were observed only in the striatum. DA contents in the hypothalamus, olfactory bulb and frontal cortex were similar to that in controls. Contents of norepinephrine and serotonin in striatum, hypothalamus, olfactory bulb and cerebral cortex were not affected by any of the treatments. Three months after MPTP alone, striatal DA recovered to 74% of controls in 8-week-old mice, whereas no recovery occurred in acetaldehyde and MPTP-treated mice. Moreover, both tyrosine hydroxylase (TH) immunocytochemistry and Cresyl violet staining showed an extensive and selective cell loss in the pars compacta of the substantia nigra (SNc) of the mice treated with acetaldehyde or ethanol and MPTP, whereas MPTP alone caused only a limited cell degeneration. PMID- 2572307 TI - Kindling of full limbic seizures by repeated microinjections of excitatory amino acids into the rat amygdala. AB - Fully developed limbic seizures were kindled by repeated (every second day) microinjections of an L-glutamate plus L-aspartate (Glu/Asp) mixture (1:3 ratio; 1.5 mumol total dose). Glu alone (1.5 mumol) or Asp alone (1.5 mumol), into the rat amygdala. This excitatory amino acid (EAA)-induced kindling was durable, persisting for at least several months, and showed strong positive transfer to electrical kindling. Fully EAA kindled seizures were inhibited by focally applied NMDA-receptor antagonists. EAA kindling and electrical kindling are shown to have many similar properties. This strongly suggests that they may also have neurochemical mechanisms in common. These results further highlight the important role of EAAs in basic mechanisms involved in the generation and expression of epilepsy. PMID- 2572306 TI - Opioid peptides (DAGO-enkephalin, dynorphin A(1-13), BAM 22P) microinjected into the rat brainstem: comparison of their antinociceptive effect and their effect on neuronal firing in the rostral ventromedial medulla. AB - The highly mu-selective agonist Tyr-D-Ala-Gly-MePhe-Gly-ol-enkephalin (DAGO) produces potent, dose-dependent naloxone-reversible antinociception when microinjected into the ventrolateral periaqueductal gray (PAG) (ED50 = 0.72 nmol) or rostral ventromedial medulla (RVM) (ED50 = 0.05 nmol) as measured on the rat tail flick (TF) assay. In single-unit recording experiments, DAGO microinjected into the PAG also affected On- and Off-Cell firing in the RVM in the same way as previously demonstrated by our group for morphine. PAG-microinjected DAGO inhibits spontaneous and noxious-evoked On-Cell firing (attenuating the characteristic On-Cell burst) (n = 19), and excites spontaneous Off-Cell firing, preventing the characteristic Off-Cell pause (n = 12) at doses which suppress the TF. These results support a major role for the mu receptor in PAG and RVM mechanisms of opiate antinociception. In our experiments using BAM22P, an endogenous weakly mu-selective opioid peptide, we could not demonstrate a dose dependent antinociceptive effect, whether the peptide was microinjected supraspinally into the PAG (n = 9) or RVM (n = 11), or intrathecally at the lumbar cord (n = 4). In two animals, a naloxone-reversible antinociceptive effect was observed following the microinjection of 10 nmol BAM 22P into the RVM; however, no effect was seen in 3 animals microinjected with 20 nmol. Dyn A(1-13), a putative endogenous ligand for the kappa receptor, had no antinociceptive effect when microinjected into the ventrolateral PAG, and no effect on the firing (spontaneous or noxious-evoked) of RVM On (n = 3)- or Off (n = 2)-Cells. PMID- 2572308 TI - Forebrain afferents to the cat posterior hypothalamus: a double labeling study. AB - Using a double immunostaining technique with cholera toxin (CT) as a retrograde tracer, we examined the cells of origin and the histochemical nature of afferents to the cat posterior hypothalamus. After injection in the tuberomamillary nucleus, a number of CT-labeled cells were observed in: medial preoptic area, nuclei of the septum and the stria terminalis, amygdaloid complex, anterior hypothalamic, ventromedial hypothalamic and premamillary nuclei. CT injections in the lateral hypothalamic area gave an additional heavy labeling of neurons in: lateral preoptic area, nuclei of the diagonal band of Broca, substantia innominata, and nucleus accumbens. The posterior hypothalamus receives: 1) cholinergic inputs from the septum, the lateral preoptic area and the nuclei of the diagonal band of Broca; 2) dopaminergic afferents from A11, A13, and A14 groups; 3) histaminergic afferents from the posterior hypothalamus; and 4) peptidergic afferents such as methionin-enkephalin, galanin and neurotensin, substance P and corticotropin-releasing factor from the medial preoptic area, the nucleus of the stria terminalis and/or the posterior hypothalamic structures. PMID- 2572309 TI - Residents' corner. Answer to case of the month #2. Takayasu's arteritis. PMID- 2572310 TI - The effect of mononuclear phagocytes on the formation of spleen colonies in mice. AB - The bone marrow cells of intact CBA mice were mixed with either syngeneic bone marrow, spleen or with peritoneal macrophages or with macrophage-like J-774 cells or with human peripheral blood monocytes of healthy donors, and all mixtures were introduced into lethally X-irradiated mice. In each case, in the spleens of the recipients colonies of the predominantly myeloid type distinctly predominated. The observed effect depended on the proportion and type of the macrophages in their mixture with the bone marrow cells. Prior X-irradiation of the mice before taking their macrophages markedly decreased the observed myeloid-induced function of the peritoneal and, especially, spleen, but not bone marrow macrophages. It was found that about 50% of the CFU-S of bone marrow origin adhered to the monolayer culture of syngeneic peritoneal macrophages during 1 h of their co incubation. These CFU-S formed spleen colonies of predominantly myeloid type when they were administered to the recipients together with the macrophages acting as carriers of CFU-S. The significance and possible mechanism of the revealed function of the mononuclear phagocytes is discussed. PMID- 2572311 TI - [Changes in amino acid contents in hepatocellular carcinoma tissues]. AB - Contents of 22 amino acids in hepatoma with surrounding and distant liver parenchyma resected from 10 pathologically proven patients were determined using high performance liquid chromatography. Analysis of the results showed that the contents of total amino acids and essential amino acids in hepatoma tissues were much higher than those in the surrounding and distant liver parenchyma. The contents of 11 amino acids, including glutamic acid, asparagine, glutamine, serine, histidine, arginine, tryptophan, methionine, leucine, isoleucine and lysine were higher than those in the surrounding and/or distant liver parenchyma. There was no statistically significant difference of amino acid contents between the surrounding and distant liver parenchyma. Most amino acid contents which increased in hepatoma tissues were positively correlated with tumor volume and/or serum gamma-glutamyl transpeptidase activity. These results suggested that hepatoma tissues can selectively take up the necessary amino acids which fail to be produced by the cancer tissues as raw material for synthesis of protein. The faster the hepatoma grows, the greater the need for amino acidosis. This study may be helpful to the application of imbalanced amino acid for correction of metabolic disturbances in hepatoma patients. PMID- 2572312 TI - The content and composition of immunoreactive somatostatin in monkey pancreas. AB - A highly sensitive and specific radioimmunoassay (RIA) was developed for analyzing somatostatin (SRIF-LI). The monkey pancreas was lyophilized and extracted with 2N acetic acid. The content and composition of immunoreactive somatostatin in monkey pancreas were then evaluated by chromatography process and RIA. The concentration of SRIF-LI in monkey pancreas was around 513.8 ng/g dry weight. At least 3 components of SRIF-LI were detected in pancreatic extracts. The major component of SRIF-LI (about 92%) was SS-14, and other two were SS-28 and a form bigger than SS-28. PMID- 2572313 TI - Tacrine, a drug with therapeutic potential for dementia: post-mortem biochemical evidence. AB - A review of biochemical findings is presented which support the idea that Alzheimer's disease represents a condition for which tetrahydroaminoacridine (tacrine) may have a beneficial effect. There is evidence that clinical and histopathologic hallmarks of the disease relate to cholinergic and serotonergic dysfunction, with less obvious abnormalities in other neurotransmitters (aspartate, dopamine, gamma-aminobutyrate, glutamate, noradrenaline and somatostatin). Clinically relevant concentrations of tacrine may ameliorate the above presynaptic deficits without producing harmful (neurotoxic) effects of aspartate and glutamate. The disease seems to be associated with an early and clinically relevant degeneration of some neurons with cortical perikarya that release these amino acid transmitters. Studies are now required on the effect of tacrine on postulated harmful peptide-bond hydrolase activity within and around such cells. PMID- 2572314 TI - Prevention of malaria in travellers: a growing problem. PMID- 2572315 TI - Antihistamines. PMID- 2572316 TI - [The role of subjective factors in medication compliance]. AB - In 1974, Van Putten was wondering "Why schizophrenic patients refuse to take their medication?" Twenty years later, the entire October 1986 issue of Psychiatric Annals deals with the subject of compliance in the case of maintenance psychotropic drugs. This paper aims to relate the empirical observations made during 152 interviews with chronic psychiatric patients to the broader considerations reported in specialized literature on compliance in the case of psychotropic drugs and of lithium. These two types of data make it possible to explain how the "perceived" advantages and disadvantages of medication go beyond the positive and negative factors objectively recognized and how they intervene in the decision to continue, modify freely or give up the treatment. In conclusion, the author reports studies on the effectiveness of various strategies proposed for increasing compliance. PMID- 2572317 TI - Tardive dyskinesia: a five year follow-up. AB - A group of chronic schizophrenic patients receiving prolonged treatment with neuroleptics was assessed in 1978 at an outpatient clinic to determine the prevalence of Tardive Dyskinesia. Those with TD were reassessed after two and five years with regard to change in TD severity. The Smith scale was used every time and, on the last assessment, the AIMS scale and videotaped interviews were added. Because of the high attrition rate, results, though interesting, cannot be generalized. PMID- 2572318 TI - Drug therapy in schizophrenia--variability of outcome and prediction of response. AB - In spite of the proven benefits of neuroleptics in reducing acute psychotic symptoms and in preventing relapse in schizophrenic patients, not all schizophrenics benefit equally from neuroleptic therapy. Predictors of response include: demographics, clinical characteristics, neurologic soft signs, neurocognitive functioning, morphologic brain changes, drug blood levels, indices of blockade of the dopamine receptors, subjective response to medications as well as early symptomatic improvement. Methodological difficulties in outcome research in drug therapy are reviewed. No single factor has been identified as a reliable predictor of drug response, and it is unlikely that such a single predictor will prove useful in a heterogeneous illness such as schizophrenia. This paper reviews the factors, which have been suggested as useful in developing better understanding of variability of drug response among schizophrenics. PMID- 2572319 TI - Aberrant expression of epidermal growth factor receptor and HER-2 (erbB-2) messenger RNAs in human renal cancers. AB - Amplification, rearrangement, or overexpression of the gene for the epidermal growth factor receptor (EGFR) occurs in certain types of human neoplasia. We investigated EGFR gene structure and measured EGFR mRNA levels in human renal tumor biopsies. Seventeen renal tumors [13 renal cell carcinomas (RCCs), two Wilms' tumors, one oncocytoma, and one metastatic ganglioneuroblastoma] and their corresponding normal kidney tissues were examined for EGFR gene structural integrity by Southern blot hybridization. Twelve of these tumors (including 11 RCCs) were examined for EGFR mRNA expression levels by RNA blot hybridization. The EGFR gene was rearranged in one of 13 (8%) of the RCC specimens examined and was highly amplified in the ganglioneuroblastoma. The overall frequency of EGFR gene structure alterations in this series of renal tumors was 12%. Nine of 11 RCC specimens (82%) exhibited markedly elevated EGFR mRNA levels (approximately 2- to 6-fold). In contrast, expression of the EGFR-related protooncogene HER-2 (erbB-2) was found to be decreased in 11 RCCs and one Wilms' tumor; HER-2 gene structure, however, appeared normal in all specimens. These results indicate that overexpression of EGFR mRNA, probably due to changes in gene regulation, and underexpression of HER-2 mRNA are characteristic features of human RCC. PMID- 2572320 TI - Assignment of common allele loss in osteosarcoma to the subregion 17p13. AB - Human osteosarcomas frequently show loss of alleles on chromosome 17 as well as those on chromosome 13 that harbors the retinoblastoma gene, indicating concerted operation of another tumor-suppressing gene on chromosome 17. To assign the affected gene to a defined region of chromosome 17, we performed mitotic recombination/deletion mapping by the use of 10 polymorphic loci on chromosome 17. Of 37 tumors studied, 28 (75.7%) showed loss of heterozygosity on chromosome 17. The affected regions varied among tumors, ranging in extent from a whole chromosome to a distal segment of the short arm. However, allele loss in one region, notably in 17p13 between D17S1 and D17S30, was common to all 28 tumors, suggesting the presence of a tumor-suppressing gene in this defined region. PMID- 2572322 TI - Rapid tranquilization with neuroleptic drugs. Neurologic concerns. PMID- 2572321 TI - Alteration of pancreatic endocrine cell patterns and their secretion during pancreatic carcinogenesis in the hamster model. AB - Proliferation of endocrine cells was found to occur during early, i.e., first 12 weeks, exocrine pancreatic carcinogenesis after 6 weekly treatments of Syrian hamsters with the pancreatic carcinogen N-nitrosobis(2-oxopropyl)amine (BOP). Cells containing insulin (Ins), glucagon (Glu), and somatostatin (Som) were noted in all stages of tumor development and were present in adenocarcinomas and in metastases to the liver. Some of the cancer cells were of amphicrine (hybrid) type, i.e., produced both mucin and endocrine substances. Measurement of these hormones revealed a significant decrease in plasma Ins during early stages of carcinogenesis with concomitant increase of Ins level in pancreatic juice at 12 weeks after 6 weekly BOP treatments. Plasma Glu and Som were not changed. The changes noted, particularly in relation to Ins, suggest that proliferation of endocrine cells in pancreatic carcinogenesis may be associated with alterations in hormone secretion. PMID- 2572324 TI - Effects of bunazosin on plasma glucose levels in hypertensive diabetic patients. AB - Twenty-nine hypertensive diabetic patients received 1.5 to 4.0 mg of bunazosin daily for three months. Both systolic and diastolic blood pressures were significantly reduced by the bunazosin treatment, while heart rate was unchanged. No significant changes were noted in fasting plasma glucose levels or in levels of hemoglobin A1c. Serum lipid levels remained unchanged during treatment. It is concluded that bunazosin does not adversely affect glucose metabolism or increase serum lipid levels in hypertensive diabetic patients. PMID- 2572323 TI - Bopindolol and atenolol in patients with stable angina pectoris. Double-blind randomized comparative trial. AB - 32 patients with stable angina and a positive symptom-limited exercise test (SLET) were investigated in a double-blind randomized trial in order to assess the therapeutic efficacy of bopindolol or atenolol on the incidence of angina pectoris and on angina pectoris threshold heart rate (ATHR). After a washout and placebo period of 2 weeks, each of the two drugs were applied in dosages of 0.5 mg bopindolol and 50 mg atenolol. The dosage was increased every month up to 2 mg bopindolol and 200 mg atenolol. At the end of every period, the patients were retested by SLET. After 3 months of active treatment, we noticed that the incidence of anginal attacks was lower in the bopindolol group (2.45 vs. 3.29). The resting heart rate was also lower in the bopindolol group (55.89 vs. 63.38). No statistical significance was found between the peak work rate, ATHR, exercise duration and S-T depression. The rate-pressure product was lower in the bopindolol group. We concluded from this that bopindolol and atenolol are active in decreasing the incidence of angina, the former being more effective. Exercise performance and cardiocirculatory parameters did not differ between the two groups. PMID- 2572325 TI - [Differentiation of the type of EHF antibodies by RPHI]. AB - This article reports the use of for differentiation of the type of EHF antibodies. A total of 58 convalescent-phase sera from EHF patients in natural foci of predominant Apodemus type of EHF and in those of Rattus type of EFH were examined. In the former 34/36 (94.44%) belonged to Apodemus type, and in the latter 22/22 (100%) belonged to Rattus type. Both RPHI and HI obtained the same result in differentiation of the type of EHF antibodies. RPHI possesses sensitivity and specificity in EHF diagnosis, and may be used for differentiation of the type of EHF antibodies. PMID- 2572326 TI - Transcriptional repression of eukaryotic promoters. PMID- 2572327 TI - A single amino acid can determine the DNA binding specificity of homeodomain proteins. AB - Many Drosophila developmental genes contain a DNA binding domain encoded by the homeobox. This homeodomain contains a region distantly homologous to the helix turn-helix motif present in several prokaryotic DNA binding proteins. We investigated the nature of homeodomain-DNA interactions by making a series of mutations in the helix-turn-helix motif of the Drosophila homeodomain protein Paired (Prd). This protein does not recognize sequences bound by the homeodomain proteins Fushi tarazu (Ftz) or Bicoid (Bcd). We show that changing a single amino acid at the C-terminus of the recognition helix is both necessary and sufficient to confer the DNA binding specificity of either Ftz or Bcd on Prd. This simple rule indicates that the amino acids that determine the specificity of homeodomains are different from those mediating protein-DNA contacts in prokaryotic proteins. We further show that Prd contains two DNA binding activities. The Prd homeodomain is responsible for one of them while the other is not dependent on the recognition helix. PMID- 2572328 TI - A homeodomain substitution changes the regulatory specificity of the deformed protein in Drosophila embryos. AB - Homeodomain proteins are believed to direct developmental pathways during Drosophila embryogenesis by the specific regulation of other genes. An unresolved issue is whether it is the homeodomain or the other regions of such proteins that confer target specificity. To test the role of the homeodomain in determining target specificity, we replaced the homeobox of Deformed with the homeobox of Ultrabithorax. The resulting chimeric protein cannot activate transcription from the Deformed gene, as does the normal Deformed protein. Instead, the chimeric protein activates ectopic transcription of Antennapedia, a gene normally regulated by Ultrabithorax. Our results indicate that in the context of the developing embryo, even closely related homeodomain sequences have different target specificities. PMID- 2572329 TI - The structure of the Antennapedia homeodomain determined by NMR spectroscopy in solution: comparison with prokaryotic repressors. AB - The structure of the Antennapedia homeodomain from Drosophila melanogaster was determined by nuclear magnetic resonance spectroscopy in solution. It includes three well-defined helices (residues 10-21, 28-38, and 42-52) and a more flexible fourth helix (residues 53-59). Residues 30-50 form a helix-turn-helix motif virtually identical to those observed in various prokaryotic repressors. Further comparisons of the homeodomain with prokaryotic repressors showed that there are also significant differences in the molecular architectures. Overall, these studies support the view that the third helix of the homeodomain may function as the DNA recognition site. The elongation of the third helix by the fourth helix is a structured element that so far appears to be unique to the Antennapedia homeodomain. PMID- 2572330 TI - Powerful immunosuppression mediated by interleukin 2-activated, nonantigen specific, or H-2-restricted THY-1+ CD8+ cells. AB - Normal spleen cells cultured in high or low concentrations of interleukin (IL) 2 for 3 days contain Thy-1+ CD4- CD8+ cells that powerfully suppress primary but not ongoing or active lymphocyte responses. The precursors of these cells are Thy 1+ AGM-1- and are absent or present in greatly diminished numbers in athymic and scid mice. Suppression is neither antigen nor H-2 restricted and apparently results from reversible inactivation of resting lymphocytes. Comparable Thy-1+ CD8+ suppressor cells were also recovered from normal spleen cells cultured for 3 days with anti-CD3 antibody without added IL-2, indicating that these cells can be activated during the course of immune responses. Such cells may prevent local recruitment/activation of lymphocytes specific for new epitopes that may be expressed sequentially by proliferating tumor cells or infectious organisms. PMID- 2572331 TI - [Methodologic aspects of voltammetry]. PMID- 2572332 TI - [Study of IgE and T lymphocyte subsets in patients with serious bronchial asthma]. AB - In the present study, 30 patients with serious asthma were selected for the investigation of IgE and T lymphocyte subsets in PBL. By the indirect McAb immuno SPA assay, it was observed that the percentages of OKT3 and OKT8 but the percentage of OKT4 subset and T4/T8 ratio were significantly higher than those of normal persons (P less than 0.01). The total IgE in serum of patients (performed by ELISA) was also much higher than that of normal persons (P less than 0.05). In addition, there was a closed relationship between the level of IgE and the percentage of OKT of the patients (r = 0.37369, P less than 0.05). The results indicated that one of possible reasons of higher IgE in serum of asthma patients was the abnormality of T lymphocyte subsets in vivo. PMID- 2572334 TI - [Mechanisms of the undescended testis]. PMID- 2572333 TI - Studies on antiallergy agents. III. Synthesis of 2-anilino-1,6-dihydro-6-oxo-5 pyrimidinecarboxylic acids and related compounds. AB - A series of 2-anilino-1,6-dihydro-6-oxo-5-pyrimidinecarboxylic acids with various substituents was synthesized and evaluated in the rat passive cutaneous anaphylaxis test for antiallergic activity. High activity by intraperitoneal and oral administrations was observed for the 3-trifluoromethyl and 2-alkoxyanilino derivatives (64, 79, 81, 82 and 85). Structure-activity relationships are discussed. PMID- 2572335 TI - [Anatomy of the ectopic testis]. PMID- 2572337 TI - [Malformations associated with testicular ectopy]. PMID- 2572336 TI - [Anatomic study of bilateral ectopies and of a testis contralateral to an unilateral ectopy]. PMID- 2572338 TI - [Classification of testicular ectopies]. PMID- 2572339 TI - [Acute complications of testicular ectopy]. PMID- 2572340 TI - [Cryptorchism and cancer of the testis]. PMID- 2572341 TI - [Sterility and cryptorchism]. PMID- 2572342 TI - [Psychologic effects of testicular ectopy]. PMID- 2572343 TI - [Medical treatment of cryptorchism]. PMID- 2572344 TI - [Surgical treatment of testicular ectopy in ordinary cases]. PMID- 2572345 TI - [Treatment of upper testicular ectopies]. PMID- 2572346 TI - [Treatment of the unpalpable testis]. PMID- 2572347 TI - [Surgical treatment of bilateral ectopies]. PMID- 2572348 TI - [Treatment of anomalies associated with ectopies]. PMID- 2572349 TI - [When is it time to intervene?]. PMID- 2572350 TI - [Surgical treatment of the failed orchiopexy]. PMID- 2572351 TI - [Fertility in the ex-cryptorchid adult]. PMID- 2572353 TI - [Ethical problems related to ectopic testis]. PMID- 2572352 TI - [Prognostic factors of testicular ectopy]. PMID- 2572354 TI - Ethical issues in the treatment of cancer patients. AB - Nineteen speakers at the International Conference on Supportive Care--More than Medicine, which was cosponsored by WHO and held in Chateau Montebello, Quebec, Canada, 18-21 July 1988, presented short introductory lectures and led the Ethics Working Group's discussions on the following ethical issues relating to cancer research and the treatment of cancer patients: telling the truth; allowing to die and practice of euthanasia; clinical research; and limited resources leading to hard choices. This article presents the discussions and recommendations of this Working Group. PMID- 2572355 TI - Laboratory support for poliomyelitis eradication: memorandum from a WHO meeting. AB - This Memorandum summarizes the conclusions and recommendations of the participants at the Consultation on Strategy for Isolation and Characterization of Poliovirus Strains and Surveillance of Wild Polioviruses held in Geneva, 28-30 November 1988. PMID- 2572356 TI - Mechanism of inhibition of estrogen-induced renal carcinogenesis in male Syrian hamsters by vitamin C. AB - Dietary supplementation of vitamin C to diethylstilbestrol (DES)- or estradiol treated male Syrian hamsters is known to inhibit renal carcinogenesis by approximately 50%. To elucidate the mechanism of inhibition, the influence of administration of vitamin C on a series of previously described biochemical markers of kidney carcinogenesis was investigated. Hamsters were stratified into four groups: (i) untreated controls; (ii) vitamin C-treated; (iii) estrogen treated; and (iv) estrogen plus vitamin C-treated animals. Concomitant administration of vitamin C and diethylstilbestrol (DES) decreased concentrations of the major DES-DNA adduct by 70-90% in liver, kidney and testis than those receiving DES only. Diethylstilbestrol-4',4"-quinone has previously been shown to be the genotoxic metabolite of DES responsible for DNA adduct formation in vivo. In vitro, vitamin C reduced diethylstilbestrol-4',4"-quinone to cis- and trans diethylstilbestrol in a dose-dependent fashion. Changes in activities of quinone reductase, catalase, superoxide dismutase and of glutathione metabolizing enzymes (glutathione peroxidase, glutathione reductase, gamma-glutamyl transpeptidase and glucose-6-phosphate dehydrogenase) in response to vitamin C were not observed or not sufficiently large to account for the 50% decrease in tumor incidence. No differences were detected in indirect estrogen-induced kidney DNA adducts in response to vitamin C treatment. It is concluded that vitamin C inhibits estrogen induced carcinogenesis by reducing concentrations of estrogen quinone metabolites and their DNA adducts. PMID- 2572357 TI - Modulation of cardiac autonomic neurotransmission by epicardial superfusion. Effects of hexamethonium and tetrodotoxin. AB - The heart contains superficial cardiac nerves whose effects may be modulated by pericardial fluid bathing the epicardium. We tested this hypothesis in open-chest dogs anesthetized with secobarbital. Oxygenated normal Tyrode's solution (NT) or NT containing hexamethonium, a ganglionic blocker (500 microM), or tetrodotoxin, a blocker of axonal neurotransmission (5 microM, TTX), was instilled into the pericardial cavity to superfuse the epicardium of the whole heart. During each superfusion, effective refractory period (ERP) was determined in deep intramyocardium (greater than or equal to 4 mm in depth from the epicardium) of anterior and posterior left ventricle and in the subendocardium of the right ventricle in the baseline state and during bilateral cervical vagal stimulation (VS) or ansae subclaviae stimulation (SS). Lengthening of ERP induced by VS during superfusion with NT (6.9 +/- 0.3 msec, mean +/- SEM, n = 36) was eliminated during subsequent superfusion with hexamethonium (0.9 +/- 0.5 msec, p less than 0.001). Hexamethonium also prevented sinus arrest induced by VS but did not affect shortening of ERP induced by SS (17.3 +/- 1.3 to 16.6 +/- 1.0 msec, n = 26). TTX suppressed VS-induced changes in ERP (6.3 +/- 0.3 to 1.5 +/- 0.5 msec, n = 32, p less than 0.001) and SS-induced changes in ERP (18.8 +/- 1.6 to 6.0 +/- 0.9 msec, n = 23, p less than 0.001) but did not affect changes in ERP induced by intravenous administration of norepinephrine or methacholine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572358 TI - Significance of myocardial alpha- and beta-adrenoceptors in catecholamine-induced cardiac hypertrophy. AB - The role of alpha- and beta-adrenoceptors in the development of catecholamine induced cardiac hypertrophy in vivo was investigated. Rats received a constant intravenous infusion of norepinephrine or sodium chloride (control) for 3 days. The norepinephrine infusion was combined with the alpha-blocker prazosin, the beta-blocker metoprolol, or both blockers. For modulation of the work load of the heart, the calcium channel blocker verapamil was added to the norepinephrine infusion. A further group of animals was treated with the alpha-adrenergic stimulator norfenephrine, which also was combined with prazosin or verapamil. Norepinephrine induced significant increases in mean aortic pressure, left ventricular dP/dtmax, heart rate, and total peripheral resistance. The left ventricular weight/body weight ratio was significantly elevated and was accompanied by an increase in the RNA concentration and the RNA/DNA ratio. Prazosin as well as metoprolol partially antagonized the increase in left ventricular weight and RNA concentration, whereas simultaneous prazosin and metoprolol treatment prevented the norepinephrine-induced alterations. Although combination of norepinephrine with verapamil resulted in considerable reduction of all functional parameters, the development of cardiac hypertrophy and the elevated RNA/DNA ratio were not significantly influenced. Stimulation of alpha receptors with norfenephrine elicited an increase in total peripheral resistance and in left ventricular weight, which was abolished by prazosin. Verapamil did not affect the norfenephrine-induced cardiac hypertrophy, although it normalized essentially all functional parameters. Thus, the rapid development of cardiac hypertrophy in the norepinephrine model seems to be directly mediated by stimulation of myocardial alpha- and beta-adrenoceptors rather than by hemodynamic changes. PMID- 2572359 TI - Growth hormone-releasing hormone influences neuronal expression in the developing chick brain. I. Catecholaminergic neurons. AB - We have examined catecholaminergic expression during development in the chick embryonic brain using tyrosine hydroxylase (TH) activity as a biochemical marker for catecholaminergic neurons. TH activity was detectable as early as after 4 days of incubation in whole brain homogenates and increased throughout embryonic development. The greatest increase in enzyme activity was observed between embryonic days 8 and 15, a period of active neuronal maturation and synaptogenesis. Growth hormone-releasing hormone (GHRH) was tested for its influence on TH activity during embryonic development. Eight-day-old embryos that received GHRH (50 ng/50 microliters) in ovo on days 1, 3, 5 and 7 exhibited a significant (P less than 0.001) increase in TH activity. Similar results were obtained when GHRH was administered in a single 50 ng/50 microliter dose on day 1 or day 3 of development. However, embryos receiving the same dose of GHRH on day 5 exhibited no significant difference in TH activity as compared to controls. When growth hormone (GH, 100 ng/50 microliters) was administered during the same critical period (day 3) no difference was observed in TH activity as compared to controls. Thus, the effects of GHRH on TH activity do not appear to be mediated through GH. We interpret these data to mean that GHRH can enhance catecholaminergic phenotypic expression in the chick embryonic brain when administered during a discrete critical period of development from days 1 to 3 of embryonic age. PMID- 2572360 TI - Developmental expression of the amyloid precursor protein, growth-associated protein 43, and somatostatin in normal and trisomy 16 mice. AB - The expression during development of 3 genes located on mouse chromosome 16 (MMU 16) which are implicated in neurobiological processes was examined by blot hybridization beginning at early gestational ages in the mouse. The 3 genes, amyloid precursor protein (App), preprosomatostatin (Smst), and growth-associated protein 43 (Gap43), exhibited distinct profiles of expression. App expression increased steadily throughout fetal and postnatal development. Smst expression peaked during the third postnatal week, then reached a plateau at a slightly lower level in adults, and Gap43 expression was highest in the early postnatal period, declining in adults to levels below those seen at the earliest timepoints examined. Smst message levels exhibited a 1.5-fold increase in the brains of trisomy 16 (Ts16) mice as compared to normal littermates on day 15 of gestation, while Gap43 and App message levels were elevated approximately 2-fold. PMID- 2572361 TI - Direct visualisation of the soluble pool of tubulin in the neuronal growth cone: immunofluorescence studies following taxol polymerisation. AB - Previously we have shown that the predominant form of soluble alpha-tubulin in the growth cone is C-terminally tyrosinated (Gordon-Weeks and Lang. Dev. Brain Res., 42 (1988) 156-160). Here we show that when growth cones are incubated in taxol, this soluble pool of alpha-tubulin is polymerised onto the ends of the neurite microtubules that enter the proximal part of the growth cone indicating that it is assembly-competent. PMID- 2572362 TI - Developmental profile of DNA synthesis and hydrolase activities in human fetal kidney. AB - This study provides original data on human fetal kidney developing during the 13th to 18th week of gestation. The parameters evaluated were DNA synthesis and the activities of 5 hydrolases which are considered as good markers of the brush border membrane differentiation. The conclusions are that DNA synthesis decreased slightly from the 16th to 18th week. The activities of maltase, trehalase, alkaline phosphatase and leucylnapthylamidase remained nearly stable during the studied period. Only the gamma-glutamyltransferase activity decreased significantly between the 15th and 16th week, then it returned close to the 13th week value. The current results suggest that during the 13-18 week period of gestation, cell proliferation is slowed down while maturation of some enzymic activities of the brush border are not importantly modified. The present basic data might be used as reference standards by investigators in the field of human nephrogenesis. PMID- 2572363 TI - DNA polymorphisms at fibrinogen loci and plasma fibrinogen concentration. AB - Associations have been reported between restriction fragment length polymorphisms (RFLPs) at fibrinogen loci and plasma fibrinogen concentration, in a British study. We have examined a series of unrelated Norwegians. We found no association between plasma fibrinogen concentration and any genotype in either of two fibrinogen polymorphisms examined (one at the alpha-fibrinogen locus, the other at the beta-fibrinogen locus). We have also examined monozygotic twins and evaluated heritability of fibrinogen level by the intraclass correlation coefficient. We arrived at an unimpressive estimate of heritability. With such a low level of heritability, it would have been surprising if we had found an association with a single gene marker in a relatively limited series of people. The reason for the discrepancy between the British and the Norwegian study is unknown. Great care has to be exercised in interpreting disease associations, since with DNA variations being examined at an increasing number of "candidate loci", the risk of finding spurious associations increases with the number of analyses conducted. PMID- 2572365 TI - Opioid receptor modulation of neural transmission in the rabbit coeliac ganglion and ganglionic opioid receptor activation by bunitrolol. AB - 1. We examined the preganglionic splanchnic nerve activity and postganglionic renal nerve activity before and after a local injection of naloxone (20 micrograms/kg) into the coeliac ganglion of anaesthetized rabbits. This was done during graded hypertension, induced by the administration of phenylephrine (0.5 10 micrograms/kg, i.v.) and with selective intraganglionic injection of methionine-enkephalin (ME) and bunitrolol, which is a beta-blocker. 2. During hypertension both pre- and postganglionic discharge decreased, but only postganglionic discharge was inhibited by naloxone treatment into the ganglion. 3. Local injection of ME (0.1-10 micrograms/kg) into the coeliac ganglion decreased postganglionic activity by 9.0 +/- 1.0 to 41.2 +/- 4.7% from control, and this decrease was inhibited by naloxone. 4. Administration of bunitrolol (1 300 micrograms/kg) decreased postganglionic discharge by 3.9 +/- 1.4 to 39.7 +/- 2.4% of the control and this decrease was also inhibited by naloxone. 5. These results suggest that opioid receptors in the coeliac ganglion play an inhibitory role in neural ganglionic transmission and that this inhibitory action reduces postganglionic sympathetic discharge. PMID- 2572364 TI - Actions of some autacoids and peptides, including relaxin, on costo-uterine muscle from rats. AB - 1. The actions of angiotensin II, bradykinin, oxytocin, arginine vasopressin, relaxin, serotonin and the prostaglandins E2 and F2 alpha were examined on preparations of costo-uterine muscle from stilboestrol-treated rats. 2. All the agonists, except relaxin, when used in concentrations which contract the rat uterus, also produced contractions of costo-uterine muscles. Concentration response curves were steep and maximal responses to the agonists were comparable. The negative log molar EC50 values were: serotonin, 6.5; angiotensin II, 8.8; bradykinin, 8.4; PGE2, 8.3; PGF2 alpha, 7.1. The EC50 values (units/L) for oxytocin and vasopressin were 4.4 and 2.7 respectively. 3. Indomethacin (2.8 or 5 mumol/L) did not decrease the contractile effects of the peptides or serotonin. The effects of serotonin were reduced, but not reversed, by methysergide (0.94 mumol/L). 4. Porcine relaxin inhibited field stimulation-induced contractions of costo-uterine muscle and uterine horns from immature rats pretreated with oestradiol cypionate and from stilboestrol-treated mature rats. It was much less potent, and its effects were less clearly concentration-related, on costo-uterine muscle. 5. The inhibitory effects of relaxin on the uterus were unaffected by propranolol (1 mumol/L), confirming that on this tissue relaxin acts independently of the release of catecholamines. Progesterone (30 mumol/L) was also without effect on the action of relaxin on the uterus. 6. These results taken together indicate that the costo-uterine muscle of the rat: (i) contracts in response to serotonin and the peptides angiotensin II, arginine vasopressin, bradykinin and oxytocin independently of the release of the contractile prostaglandins F2 alpha and E2; and (ii) in contrast to the uterus, may lack a significant population of receptors for relaxin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572366 TI - Acute changes in the properties of baroreflexes in man after beta-blockade. AB - 1. To clarify whether acute changes in the properties of baroreflexes can occur in man, we evaluated the time course of baroreflex control of heart rate and cardiopulmonary baroreflex control of forearm vascular resistance (FVR) over 240 min after intravenous administration of propranolol (0.2 mg/kg) in 13 healthy young men. 2. Systolic and diastolic blood pressure remained unchanged after propranolol. Propranolol significantly decreased cardiac index and heart rate, and significantly increased total peripheral resistance. These effects remained unchanged for 240 min after propranolol. 3. Baroreflex control of heart rate was significantly augmented immediately after, and at 30, 60 min after propranolol, but partly reverted to the initial level afterwards. Cardiopulmonary baroreflex control of FVR was reduced immediately after, and at 30, 60 min after propranolol, but partly reverted to the initial level afterwards. Pressor responses to phenylephrine was reduced immediately after propranolol, but no significant differences were observed after 30 min. 4. These results suggest that acute changes in the properties of baroreflexes occur in man after propranolol. PMID- 2572367 TI - Alpha-agonist modifies baroreflex vasoconstriction by a postjunctional mechanism in dogs. AB - 1. We examined whether or not circulating alpha-agonist modified baroreflex vasoconstriction of the hindlimb, using anaesthetized dogs in which the limb was vascularly isolated and perfused with blood from a donor dog using a pulsatile pump. 2. The open-loop gain (G) of the baroreflex was estimated from changes in mean arterial pressure following mild quick haemorrhage from the aorta of the recipient dog. 3. The hindlimb perfusion pressure increased after haemorrhage due to neurogenic vasoconstriction. 4. An overall gain (Gh) of the baroreflex hindlimb vascular bed control system was estimated from the ratio of the increase in hindlimb perfusion pressure to the change in systemic arterial pressure of the recipient dog. 5. Administration of a relatively selective alpha 1-agonist with no prejunctional beta 2 stimulating action (phenylephrine) or a selective alpha 2 agonist (clonidine) to the donor dog increased its systemic arterial pressure and augmented Gh. 6. Since both drugs were administered to the donor dog and could not enter into the recipient dog, these drugs did not affect the recipient's sympathetic nervous system, including the central nervous system and afferent limb of the baroreflex system. Therefore, these drugs could modify baroreflex vasoconstriction of the hindlimb only at the junction of the efferent sympathetic nerve and the vascular smooth muscle. 7. It was concluded that postjunctional alpha-adrenoceptor stimulation augments neurogenic vasoconstriction. PMID- 2572368 TI - Peripheral lymphocyte subsets and their responsiveness in human strongyloidiasis. AB - Surface phenotypes and proliferative responses of peripheral lymphocytes were investigated in a total of 64 patients with strongyloidiasis. A significant increase of CD4+ and OKIa1+ cells and a relative decrease of CD8+ cells were observed among the patients. A considerable rise of spontaneous mitogenesis and an enhanced ability to produce interleukin 2 of peripheral lymphocytes were also found in many patients. On the other hand, lymphoproliferative responses to mitogens were found to be significantly lower in the strongyloidiasis patients than in the controls. The possible relationships among these observations were discussed. PMID- 2572369 TI - Detection and clinical implication of anti-neutrophil cytoplasm antibodies in Wegener's granulomatosis and rapidly progressive glomerulonephritis. AB - The specificity of anti-neutrophil cytoplasm antibodies (ACPA/ANCA) was investigated in patients suffering from Wegener's granulomatosis (WG), various forms of systemic diseases including vasculitides (non-WG), and different types of biopsy-proven glomerulonephritides. In particular, the diagnostic significance of ACPA/ANCA was assessed in patients affected by rapidly progressive glomerulonephritis with and without systemic manifestations. From 25 patients with active Wegener's granulomatosis 22 showed the classical diffuse finely granular cytoplasmic staining of neutrophils as did 4/31 patients with idiopathic rapidly progressive glomerulonephritis. One patient with biopsy confirmed Wegener's granulomatosis, six patients with microscopic polyarteritis and 3/31 patients with idiopathic rapidly progressive glomerulonephritis showed a focal cytoplasmic staining exhibiting a rosette-like pattern. One further patient with Wegener's granulomatosis displayed a prominent fluorescence of the outer nuclear membrane as well as a punctate-diffuse nuclear staining resembling that of granulocyte specific antibodies. Another patient with active Wegener's granulomatosis did not react in the ACPA/ANCA-test. These findings demonstrate different staining patterns of neutrophils which are related to different clinical entities within the spectrum of small vessel vasculitis. Moreover, they point out that different antigens are involved in the various types of vasculitis. In classical cases of Wegener's granulomatosis, but not in other forms of vasculitis, the titer of ACPA/ANCA showed a close relationship to the number of organs involved. PMID- 2572371 TI - Alcohol, smoking and body build: obesity as a result of the toxic effect of 'social' alcohol consumption. AB - Previous work showed that obesity in the average human male is not due to increased caloric intake. To test the hypothesis that 'social' ethanol consumption causes obesity by a hepatotoxic mechanism, the relationships between alcohol intake, cigarette smoking, serum gamma-glutamyl transpeptidase (GGT) and body build were investigated in 816 adult patients, 491 males and 325 females. A large part of the Broca index variance could be explained by hepatic damage as reflected by the GGT level. The higher the GGT, the more overweight were the subjects. Hyperinsulinemia may be the pathogenetic link; insulin is the strongest known blocker of lipolysis. Almost the total variation of obesity with GGT, however, occurred in the range of GGT up to 25 U/l, which is usually, but nevertheless erroneously, considered to be the normal range. This effect was independent of sex and age. Normal GGT is below 10 U/l, which is found on average in females aged less than 20 years. Females tolerate less alcohol than males. Although GGT is as high in females as in males around age 30, males drink about three times as much ethanol. For the same GGT the Broca index is significantly higher in females than in males. GGT generally increases with age; maximum GGT is reached in females in the age group 21-40 years (due to the change in drinking habits around 1968), declining thereafter; in males at age 50. Obesity per se is not correlated with a high GGT. In the females there are hormonal factors influencing obesity. Although in the females GGT decreases on average after age 40, obesity increases (due to the decrease in estrogens). After age 50 ethanol tolerance in males decreases: they reduce their alcohol consumption, and yet the GGT remains high. -Cigarette smoking is a factor which independently influences obesity. Although people who smoke tend also to drink more alcohol, smokers are significantly leaner than nonsmokers. On average males smoke about twice as heavily as females; this contributes to the fact that on average males are leaner than females despite their higher alcohol consumption. Due to lower consumption the influence of ethanol and smoking on body build is smaller in females than in males. PMID- 2572370 TI - Erythrocyte cation transport in obesity, hypertension, and during antihypertensive drug therapy. AB - The number and activity of erythrocyte ATPase-dependent sodium-potassium pump units were increased in obese subjects (p = 0.02). No link was observed between the number or activity of the pump units and hypertension. The ouabain insensitive rubidium (i.e. potassium) transport was not associated with relative body weight or blood pressure status. Sodium-lithium countertransport correlated significantly with obesity but not with blood pressure status. In the hypertensive patients, before or after therapy with verapamil, hydrochlorothiazide, pindolol or atenolol there were no significant differences in cation transport. We propose that the correlation between obesity and essential hypertension cannot be explained by these two cation transport systems. PMID- 2572372 TI - Effect of almitrine on hypoxic ventilatory drive measured by transient and progressive isocapnic hypoxia in normal men. AB - 1. In a double-blind placebo-controlled study, we have investigated the effect of the peripheral chemoreceptor stimulant drug almitrine bismesylate on hypoxic ventilatory drive (expressed as the slope of the minute ventilation/arterial oxygen saturation relationship in litres min-1+-1) as measured by both progressive isocapnic hypoxia at rest and transient hypoxia (three breaths of 100% N2) during moderate exercise, in seven normal men, to determine if the ventilatory response to the transient hypoxic stimulus is a more specific measure of peripheral chemoreceptor sensitivity to hypoxia. 2. Hypoxic ventilatory drive measured using progressive isocapnic hypoxia ranged from -0.13 to -2.65 litres min-1%-1 after placebo and from -0.20 to -6.48 litres min-1%-1 after almitrine. The response was greater after almitrine in six of the seven subjects, and the difference was significant for the whole group (P less than 0.05). 3. Hypoxic ventilatory drive measured using transient hypoxia ranged from -0.19 to -1.59 litres min-1%-1 after placebo and from -0.09 to -1.62 litres min-1%-1 after almitrine. The response was not consistently greater after almitrine, and the difference was not significant for the group. 4. Difficulties in accurately quantifying a brief rise in minute ventilation after transient hypoxia, particularly in subjects with a low hypoxic ventilatory drive, may have masked small changes in the slope of the minute ventilation/arterial oxygen saturation relationship with this method.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572373 TI - Clozapine: an atypical antipsychotic agent. AB - The pharmacology, pharmacokinetics, clinical efficacy, adverse effects, dosage, and cost of the atypical antipsychotic drug clozapine are reviewed. Clozapine is a dibenzazepine compound chemically similar to loxapine but with a distinct pharmacologic profile. Unlike currently available medications, clozapine has a low potential for causing extrapyramidal symptoms and does not induce dopamine type 2 receptor hypersensitivity. It shows affinity in vitro not only for dopamine type 1 and 2 receptors but also for histamine type 1, alpha-adrenergic type 1 and 2, serotonin type 2, and muscarinic receptors. Clozapine given orally is nearly completely absorbed and readily metabolized. Urinary excretion is the major route of metabolite elimination. Clozapine has been used to treat schizophrenia, nonschizophrenic psychotic states, depression, neuroses, and behavioral disorders. Double-blind comparative studies have shown clozapine to be superior to haloperidol, chlorpromazine, and placebo in treating the symptoms of schizophrenia, as measured with validated psychiatric rating scales. Adverse effects include orthostatic hypotension, tachycardia, benign hyperthermia, hypertension, seizures, and sedation. Many of these effects are transient. Because of the risk of agranulocytosis, a comprehensive case-management system has been developed. In treating acute psychosis, the optimum dosage of clozapine is 300-450 mg/day given orally in divided doses. The high cost of clozapine may be offset by improved patient response and reduced hospital costs. Clozapine may be superior to other agents in the treatment of refractory schizophrenia and is associated with a negligible incidence of extrapyramidal symptoms. PMID- 2572374 TI - [Nicardipine, alone or in combination with enalapril, in the therapy of arterial hypertension secondary to chronic nephropathy]. AB - The aim of the study was to evaluate the long term antihypertensive effect of nicardipine in hypertensive patients with chronic renal disease. Eight patients (creatinine clearance ranging from 51 to 78 ml/min/1.73 m2) received nicardipine (20 mg t.i.d.). Four weeks later, patients with diastolic blood pressure greater than or equal to 90 mmHg in recumbent position, were given enalapril (10 mg/day) as well. Blood pressure control was achieved in 3 patients treated with nicardipine alone and in 5 patients on a combined nicardipine-enalapril regimen, and it was maintained throughout the whole trial period (52 weeks). In two cases serum creatinine rose from 2.3 to 3.3 and from 1.4 to 2.2 respectively. However, the slope of the creatinine ratio, plotted against time, showed a significant reduction in renal function loss as compared to expected values. In conclusion, nicardipine, alone or in combination with enalapril, is an effective and well tolerated drug for use in treatment of hypertension secondary to chronic renal disease. PMID- 2572375 TI - Drug therapy for juvenile arthritis. AB - Juvenile arthritis includes a variety of rheumatic disorders with varied articular and extraarticular manifestations. The heterogeneity of JA cannot be overemphasized in a rational approach to therapy. Fortunately, most children with JA improve significantly with first-line treatment, especially those with pauciarticular disease. Only a small number of these patients go on to require second-line treatment, usually because of the evolution of their condition into the polyarticular type. Patients with polyarticular, RF-positive disease and patients with early-onset polyarthritis in association with systemic-onset disease both have poor prognoses in terms of ultimate joint function, and should receive second-line treatment early in their therapy. Injectable gold is the drug of choice for the former patients, if an initial trial of NSAIDs has failed. Patients with polyarticular onset, RF-negative disease generally have a better prognosis than other patients with polyarticular involvement. One may wait considerably longer before introducing a second-line agent for these patients. An antimalarial agent may be a reasonable choice for these patients, especially those with limited polyarticular involvement (5 to 10 active joints). This latter principle may also be applied to patients who evolve from pauciarticular to polyarticular involvement. In all such cases, penicillamine is almost never used as first choice. If the patient is a young child, the difficulty with injecting gold and the hazards of antimalarial agents may limit the use of these treatments. In situations such as this, auranofin or sulfasalazine may be the drug of choice. Cytotoxic/antimetabolic therapy should be reserved for those children who have continued active disease despite full courses of two SAARDs, or for those who have been unable to take SAARDs because of their side effects. Corticosteroid therapy should be used only when specifically indicated, and every attempt should be made to wean the patient from it as soon as feasible. The aim of therapy is to ensure that patients enter their remission in the best condition possible. For this to be assured, a sensible approach to drug therapy is mandatory. PMID- 2572377 TI - In vivo and in vitro metabolism of benzo(a)pyrene by the larva of the newt, Pleurodeles waltl. AB - 1. The larva of the amphibian species, Pleurodeles waltl was shown to metabolize benzo(a)pyrene in vivo into a variety of oxidized products. 2. In vitro, BaP hydroxylase (AHH) activity was found in hepatic microsomes and postmitochondrial fractions from both larvae and adults of the pleurodele. 3. The clastogenic effect of BaP formation of micronuclei in the erythrocytes was shown to be related to the presence of BaP quinones in the tissues of the newt. PMID- 2572376 TI - A comparative study on acetyl-CoA synthesising enzymes in spinal cord from cows, horses and pigs. AB - 1. Comparative data are presented of the activities of pyruvate dehydrogenase complex and acetyl-CoA synthetase and of the acetate content in homogenates from ventral grey matter in spinal cord from cows and two non-ruminant species, pigs and horses. The methods used in the study are evaluated and discussed. 2. The total pyruvate dehydrogenase complex activity was 24.9-29.9 mU/mg protein and did not differ between the species. The part of the complex that was in active form at the sampling occasion was 60, 85 and 95% in cows, pigs and horses, respectively. 3. Acetyl-CoA synthetase activity differed significantly between the species and was 0.93, 1.28 and 2.61 mU/mg protein in pigs, cows and horses, respectively. The highest cytosolic activity was found in the horses. Acetate concentration at half maximal reaction velocity (at saturating CoA and ATP levels) was found to be 0.15-0.70 mM and did not differ between the species. 4. Acetate content was 63, 83 and 96 micrograms/g wet wt in cows, horses and pigs, respectively. 5. It is concluded that there seems to be no striking difference in acetyl-CoA synthesis in peripheral nerves between ruminants and non-ruminant species. PMID- 2572378 TI - In vitro inhibition of prostaglandin H synthase by compounds from the exocrine secretions of lace bugs. AB - 1. Selected lace bug-derived and related compounds were shown to be in vitro inhibitors of mammalian and insect derived prostaglandin H synthase. 2. Two compounds, 2,6-dihydroxyacetophenone and 2,4,6-trihydroxyacetophenone, were significantly better inhibitors of prostaglandin synthesis than aspirin, whereas 2-nonyl-5,7-dihydroxychromone and 1-(2,6-dihydroxyphenyl) dodecan-1-one were equivalent to aspirin. 3. 2,4-Dihydroxyacetophenone was less effective in inhibiting the prostaglandin H synthase and 2-nonyl-5-hydroxychromanone showed no inhibition. 4. Three compounds, 2,6-dihydroxyacetophenone, 2,4,6 trihydroxyacetophenone and 1-(2,6-dihydroxyphenyl) dodecan-1-one were equal to aspirin in PSI inhibition with fat body preparations of the American cockroach Periplaneta americana L. PMID- 2572379 TI - Pharmacological actions of a new class of neuropeptides, the leucokinins I-IV, on the visceral muscles of Leucophaea maderae. AB - 1. Leucokinins I-IV did not activate visceral muscles uniformly as a class but rather showed a selective action on the muscles of the hindgut. This organ showed a contractile response to all of the leucokinins at 3 x 10(-10) M that was 5-10% above the mean level of spontaneous activity. The maximum response for each peptide was recorded at 2.1 x 10(-7) M. 2. Both the foregut and the oviduct were 100-1000 fold less sensitive than the hindgut, and each of the former organs required more than 10(-8) M to elicit a detectable excitation. The heart, by comparison, failed to give consistent responses with any of the peptides. 3. The leucokinins caused a protracted excitation of contractile events in the hindgut that lasted for more than 60 min. Moreover, all four peptides evoked contractions from hindguts after membrane depolarization with 158 mM potassium. 4. This result shows that nonsynaptic receptors for the peptides exist in visceral muscle. The leucokinins showed no evidence of facilitating the reentry of calcium into calcium depleted hindgut preparations. PMID- 2572380 TI - Comparative metabolism of benzo[f]quinoline by liver microsomes from brown bullheads and rats. AB - 1. Liver microsomes from rats were considerably more active in metabolizing benzo[f]quinoline (B f Q) than those from brown bullheads (Ictalurus nebulosus). 2. The main B f Q metabolites formed by both rat and brown bullhead liver microsomes were qualitatively similar and included B f Q-7,8-dihydrodiol, B f Q 9,10-dihydrodiol, B f Q-N-oxide, 7-hydroxy B f Q, and 9-hydroxy B f Q. 3. The liver microsomes from control brown bullheads and rats metabolized B f Q primarily at the 7,8-and 9,10-positions, respectively, whereas in the case of microsomes from 3-methylcholanthrene (3-MC)-treated rats or brown bullheads, the major site of metabolic attack was the 7,8-position. 4. A 3-MC-type of cytochrome P-450 appears to be primarily responsible for the oxidation of B f Q by control brown bullhead liver microsomes, whereas a phenobarbital-inducible type of cytochrome P-450 seems to be involved in the metabolism of B f Q by control rat liver microsomes. PMID- 2572381 TI - A comparison of norepinephrine- and ACTH-stimulated lipolysis in white and brown adipocytes of female rats. AB - 1. Isolated white and brown adipocytes (WFA and BFA) from the rat were compared with respect to their lipolytic responsiveness towards norepinephrine (NE) and adrenocorticotrophic hormone (ACTH). 2. NE yielded a Km value of 702.7 +/- 30.6 nM for WFA and 142.5 +/- 7.2 nM for BFA. The maximum lipolytic response (Vm) was 145.7 +/- 1.2 nmol glycerol/micrograms DNA/90 min for WFA and 23.7 +/- 0.2 nmol glycerol/micrograms DNA/90 min for BFA. 3. ACTH yield Km values of 31.6 +/- 1.5 and 31.9 +/- 3.1 nM for WFA and BFA, respectively. Vm values of 141.9 +/- 1.0 and 34.2 +/- 0.5 nmol glycerol/micrograms DNA/90 min were observed for WFA and BFA, respectively. PMID- 2572382 TI - [3H]serotonin binding to blastula, gastrula, prism, and pluteus sea urchin embryo cells. AB - 1. The presence of serotonin binding sites in blastula, gastrula, prism, and pluteus embryos of the sea urchin, Arbacia punctulata, was investigated by the binding of radiolabelled serotonin to dissociated embryo cells. 2. [3H]serotonin binding sites were identified in prism, early pluteus, and advanced pluteus larvae, but not in blastula or gastrula embryos. 3. The ontogeny of [3H]serotonin binding activity closely parallels that of serotonin content as previously reported in Paracentrotus lividus embryos (Toneby, 1977a). 4. Results of this study support a regulatory role of serotonin in developmental processes in postgastrula sea urchin embryos. PMID- 2572383 TI - Muscarinic receptors modulating acetylcholine release from insect synaptosomes. AB - 1. Cholinergic synapses in the central nervous system of insects contain inhibitory muscarinic receptors whose stimulation by agonists leads to a diminished output of acetylcholine; antagonists, like atropine, facilitate acetylcholine release. 2. The receptors involved appear to be of the M2-subtype. Upon activation of presynaptic receptors a significant reduction of the intrasynaptosomal cyclic AMP level as well as a significantly increased membrane potential was observed. 3. The observed membrane hyperpolarization is apparently not a consequence of a lower cyclic AMP level, thus both effects may offer alternative or synergistical mechanisms for modulating transmitter release. PMID- 2572384 TI - Evidence for a lower oesophageal sphincter in the guinea-pig. AB - 1. In vitro balloon pull-through experiments have been used to identify the guinea-pig lower oesophageal sphincter (LOS). 2. Histologically, the LOS forms a 1-2 mm ring of smooth muscle at the distal termination of the oesophagus, immediately adjacent to the gastric sling muscle. 3. Tetrodotoxin (10(-6) M) sensitive, guanethidine (10(-6) M) insensitive "on" relaxation of circular LOS muscle strips was evoked by electrical field stimulation (ES). 4. ES evoked atropine (10(-6) M) sensitive "on" contractions of gastric sling and fundus smooth muscle strips. 5. Following cessation of ES a partially atropine-sensitive "off" contraction was observed in all the smooth muscle strips. 6. The predominant response of the LOS to ES was relaxation. PMID- 2572386 TI - [3H]kainic acid binding sites in chick cerebellar membranes. AB - 1. A total particulate fraction of chick cerebellar membranes, obtained by a simple method, has been found to specifically bind [3H]kainic acid. Non-neuronal tissue, like chick liver, does not show any appreciable specific binding under the same experimental conditions. 2. Specific [3H]kainic acid binding to chick cerebellar membranes increases linearly with tissue concentration, reaches the binding equilibrium almost instantaneously and is pH and temperature dependent. 3. Specifically bound [3H]kainic acid is displaced by suitable concentrations of unlabelled kainic acid, L-glutamic acid and other excitatory amino acid analogues, both agonist and antagonist. This pharmacological pattern agrees with the general pharmacological properties of kainic acid receptors. 4. Saturation kinetic studies of kainic acid binding sites show one single binding mode with an apparent dissociation constant KD = 278 nM and a maximum number of binding sites of 187 pmoles/mg of protein. 5. In view of the mentioned data and the high amount of receptor sites found in chick cerebellar membranes, as compared with related values in rat cerebellum, we suggest that these receptors play a different physiological role or that they have a different cellular localization in chick and rat cerebellum. PMID- 2572385 TI - Changes in the catecholamine metabolism in the adrenal medulla of male hamsters with experimental hyperprolactinemia. AB - 1. Prolactin (PRL) can play a role as a physiological modulator of adrenal medulla function in several rodents. 2. We have examined the effects of hyperprolactinemia induced by ectopic pituitary grafts in Syrian hamsters on the adrenal medulla contents of catecholamines (CA) and their metabolites, as well as on the activities of several enzymes involved in the metabolism of these amines. 3. Increases in the peripheral levels of PRL in these animals were associated with decreases in adrenal medulla weight and increases in adrenal medulla contents of norepinephrine, epinephrine and vanilmandelic acid, the main degradative metabolite of CA, while adrenal medulla contents of the O-methylated derivatives of CA, normetanephrine and metanephrine, were unaltered. 4. These changes were correlated with increases in the adrenal medulla activity of monoamine oxidase, while the activities of tyrosine hydroxylase, phenylethanolamine-N-methyl transferase and catechol-O-methyl transferase were unaltered. 5. These results indicate that PRL is able to act on the adrenal medulla of hamsters by increasing the ability of these cells to metabolize CA via oxidative deamination. PMID- 2572387 TI - Age dependent changes in metallothionein and accumulation of cadmium in horses. AB - 1. Analysis of livers and kidneys from 28 horses for cadmium, zinc and metallothionein showed low cadmium content in liver. There was a gradual increase in cadmium content in kidney with age. 2. Metallothionein values varied with zinc content in the liver and with cadmium content in the kidney; copper values did not vary in either tissue. 3. Metallothionein was localized mainly in the cytoplasms in liver and kidney of horses by immunohistochemistry. PMID- 2572388 TI - The actions of halothane on spontaneous activity, action potential shape and synaptic connections of the giant serotonin-containing neurone of Lymnaea stagnalis (L.). AB - 1. Cerebral giant cells (CGCs) in the isolated central nervous system (CNS) of the pond snail Lymnaea stagnalis (L.) exhibit bursting activity when superfused with anaesthetic concentrations of halothane. 2. Calcium-dependent components of the CGC action potential appear more sensitive to halothane than do other ionic mechanisms. 3. Higher concentrations of halothane block the chemical synaptic connection between CGC and buccal motoneurone B1, but have no effect on the strong electrotonic coupling between the CGCs. 4. The mechanisms underlying CGC bursting and synaptic block in the presence of halothane are discussed. PMID- 2572389 TI - Rat myometrial Na/K ATPase is increased by serum but not by isoproterenol and relaxin. AB - 1. Na/K ATPase activity in rat myometrial cells in culture exhibited a Kapp of 0.93 mM for Rb+ and a Ki of 31 microM for ouabain with respect to Rb+. 2. 86Rb+ uptake was stimulated by serum and monensin but was not affected by the uterine relaxants isoproterenol and relaxin in 0.5-7.5 mM Rb+. Nonetheless, these relaxants elicited significant increases in 45Ca2+ efflux under similar conditions. 3. These data suggest that increased Na/Ca exchange resulting from a stimulation of Na/K ATPase is not involved in the mechanism of action of relaxin and isoproterenol in the uterus. PMID- 2572390 TI - Effects in vitro of copper and zinc on hepatic cytochrome P-450 activities. AB - 1. In previous studies we have shown that hepatic copper and zinc increases and liver microsomal cytochrome P-450 activities greatly decreases in adjuvant arthritic rats. 2. In the present paper we study if the changes in copper and zinc could be related to depression of drug microsomal activity. Thus, the effect of in vitro addition of copper or zinc to microsomal fraction upon aminopyrine N demethylase (AND) and aniline p-hydroxylase (APH) activity was measured. 3. Both metals produced an inhibition of enzyme activity. The reduction of AND and APH activities produced by copper (ID25 = 4.7 x 10(-5)M to AND; 1.05 x 10(-5)M to APH) was greater than that obtained with zinc (ID25 = 2.26 x 10(-4)M to AND; 3.3 x 10(-4)M to APH). PMID- 2572392 TI - Occurrence of various forms of metallothionein in the rat after a short-term cadmium injection regimen. AB - 1. Results are presented showing that the metal-binding metallothionein (Mt) species induced in rat liver in response to Cd administration consist of dimeric and trimeric forms of Mt. A monomeric form might be the first step in the polymerization process. 2. Two proteins (about 8 and 20 kDa mol. wt) are found in hippocampus, but not in brain cortex. 3. These proteins could not be demonstrated to cross-react with our Mt antibody, but the largest of them has strong Cd binding capacity. 4. Our Mt antibody cross-reacts with a high metal affinity protein present in both brain cortex and hippocampus of twice the mol.wt (20 kDa) of our purified rat liver Mt standard. 5. The results indicate, however, that these Mt like proteins probably emerge from high molecular or membrane bound forms in the cells. 6. A theory is proposed that the predominant polyacrylamide gel band, matching the monomeric, rat liver Mt standard band, seen for all tissues studied in the present work originate from two sources, namely membrane bound and heavy metal induced monomeric form. 7. It is furthermore suggested that those tissues playing an active role in heavy metal metabolism and in protection against toxicity of such metals contain soluble Mts whose active metal-binding forms are oligomers. PMID- 2572391 TI - New differences between the Wistar rat and Octodon degus, a putative laboratory animal resistant to morphine. AB - 1. The effects of CNS depressants (methadone and alcohol) and natural neurotransmitters (NA and ACh) are studied in O. degus. 2. O. degus shows resistance to methadone in the formalin algesiometric test and EEG. 3. Ethanol elimination profile suggest the presence of an atypical alcohol dehydrogenase in O. degus 4. O. degus is extremely resistant to the pressor effects of noradrenaline 5. the isolated atrium of this rodent is 40 times more sensitive to the negative chronotropic effect of methadone, than the rat atrium. 6. These effects could be explained in terms of an important catecholamine and endorphin co-secretion from adrenal glands in O. degus. PMID- 2572393 TI - Effects of a novel perfluorochemical emulsion on lymphoid tissues and immunocompetence in rats: time course effects relative to immunological challenge. AB - 1. The effects of intravenous (i.v.) injection of low doses of a novel perfluorochemical (PFC) emulsion on lymphoid tissues and antibody production against i.v.-injected sheep red blood cells (SRBC) have been studied relative to the timing of immunization in rats. 2. Spleen and, to a lesser extent, liver weights were significantly increased in response to injection of emulsion but no consistent pattern was observed. 3. Thymus weight was consistently decreased following injection of emulsion; in contrast, mesenteric lymph node (MLN) weights were unchanged throughout except for a significant increase in animals injected with emulsion 24 hr prior to immunization. 4. The mean plasma antibody titre to SRBC showed a variable response in animals receiving the emulsion: titres were significantly increased in rats injected with emulsion 1 hr prior to immunization whereas they were significantly reduced in animals injected with emulsion 7 days before SRBC. 5. These results show that lymphoid tissue weights and plasma antibody titres to SRBC vary according to the time of a previous or subsequent injection of PFC emulsion. PMID- 2572394 TI - Schistosoma mansoni: reversible destruction of testes by procarbazine. AB - 1. The anticancer drug procarbazine is profoundly damaging to the testes of Schistosoma mansoni when administered at 200 mg/kg or more to the mouse host. Somatic tissues appear entirely unaffected. 2. Within 2 days the meiotic process is disrupted, and primary and secondary spermatocytes and spermatids are destroyed and replaced by amorphous granular material. 3. The testes regenerate within about 15 days, apparently from surviving spermatogonial resting cells near the germinal epithelium of the testis. 4. Livers of mice treated once 7 weeks earlier have numerous egg granulomas and give rise to many miracidia, suggesting that full testis function is regained. 5. Male worms given the drug at 200 mg/kg at 19 days of age and fixed 7 weeks later have on average one testis less than control worms, indicating that about 15% of immature testes are unable to regenerate, whereas mature worms given the drug regenerate the normal number of testes. 6. The drug did not have significant antispermatogenic effects when male worms were incubated for a week with various concentrations in vitro, suggesting that a host metabolite is the active agent. PMID- 2572395 TI - Teratogenicity of combinations of sodium dichromate, sodium arsenate and copper sulphate in the rat. AB - 1. The teratogenicity of sodium dichromate (2 mg Cr/kg), sodium arsenate (5 mg As/kg) and copper sulphate (2 mg Cu/kg) in female Wistar rats was studied following the administration of the test compounds separately and in their various combinations on gestation day 8. 2. The test compounds administered separately and the combination of dichromate plus Cu2+ were non-fetotoxic and either non- or weakly teratogenic, whereas arsenate/Cu2+ was both weakly fetotoxic and teratogenic. 3. Dichromate/arsenate and dichromate/arsenate/Cu2+ caused a marked decrease in mean fetal weight and an increased incidence of fetal resorption and abnormality formation. PMID- 2572396 TI - Further characterisation of the dopamine-inhibitory receptor in Helix and evidence for a noradrenaline-preferring receptor. AB - 1. The cells in this study responded with a hyperpolarization to the following agents in this order of potency; dopamine greater than noradrenaline phenylephrine = octopamine. 2. 6,7 ADTN had a relative potency of 0.1 compared to dopamine. 5,6 ADTN did not inhibit the cells in this study. 3. The D1 receptor agonists SKF38393 and dihydroxynomifensine mimicked the effect of dopamine on these cells but were over 100 times less active, whereas the D2 selective agonists quinpirole and RU24213 were without effect. 4. Both the D1 antagonist SCH23390 and the D2 antagonist sulpiride antagonised the dopamine response with pA2 values of 6.1 and 6.7, respectively. 5. Five cells that responded to dopamine with a hyperpolarization were depolarized by noradrenaline. The order of potency of compounds at eliciting this depolarization, noradrenaline greater than phenylephrine greater than octopamine indicated that this response may be mediated by a noradrenaline-preferring receptor. PMID- 2572397 TI - Lack of platelet monoamine oxidase activity in Cebus monkeys (Cebus albifrons). AB - 1. Recent evidence suggests that monoamine oxidase (MAO) plays an important role modulating the extrapyramidal syndromes produced by neuroleptic drugs in both human and nonhuman primates. 2. To evaluate the possibility of using peripheral blood platelet MAO-B levels as indices of central nervous system MAO-B effects, we measured platelet MAO-B levels in Cebus monkeys that were previously tested with neuroleptics (N = 36) or drug naive (N = 6). 3. No platelet MAO-B was consistently detectable in these blood samples. 4. Thus platelet measures of MAO B do not reliably reflect brain MAO-B function in nonhuman primates and do not offer a useful model for studying blood-brain MAO-B relationships. PMID- 2572398 TI - Reconstitution of scid mice by injection of varying numbers of normal fetal liver cells into scid neonates. PMID- 2572399 TI - Reconstitution of lymphocyte subsets in scid mice by transplantation of fetal primordia. PMID- 2572400 TI - Establishment of assays for human hematopoietic cells in immune deficient mice. AB - Our knowledge of the organization and regulation of the murine hematopoietic stem cell hierarchy has, in large measure, been the result of the development of in vivo reconstitution assays for cells of various levels of differentiation. For example, many basic stem cell concepts were established using CFU-S as a paradigm. Although there are no clonal assays for the earliest pluripotential cells, they can nevertheless be quantitated based on their ability to reconstitute animals with a deficient hematopoietic system. A similar understanding of the human hematopoietic system has lagged because of the lack of a suitable assay for human stem cells. Using several different approaches it has become possible to transplant human hematopoietic cells into immune-deficient mice. Since both lymphoid and myeloid cells have been engrafted, these experiments may lay the foundation for an assay for human pluripotent stem cells. Moreover it should now become possible to establish experimental animal models of a large number of human hemopathies including leukemia. PMID- 2572401 TI - Linkage analysis of human chromosome 4: exclusion of autosomal dominant retinitis pigmentosa (ADRP) and detection of new linkage groups. AB - As part of our ongoing linkage studies of degenerative retinal diseases, we tested seven DNA markers and two classical genetic markers from chromosome 4 in two extended families with autosomal dominant retinitis pigmentosa (ADRP). Our goals were (1) to detect or exclude linkage of ADRP to markers spanning most of chromosome 4 and (2) to contribute useful new information regarding the linkage map of this chromosome. Our results exclude linkage of ADRP from more than 82% of chromosome 4. We detected four new linkage relationships: loose linkage of K082 (D4S10) and G1E5 (D4S21) at a distance of 21 cM; loose linkage of 4F2 (D4S18) and GC protein at a distance of 19 cM; tight linkage (i.e., no recombinants) between B3D (D4S44), B5A (D4S40), and the MNS blood group; and tight linkage between 4F2 and GDS5 (D4S23). These data, combined with previously reported data, exclude ADRP from approximately 35% of the human genome. PMID- 2572402 TI - The HLA class I locus: analysis of RFLPs in hereditary hemochromatosis. AB - The gene for hereditary hemochromatosis is linked to the HLA locus on chromosome 6. Four cloned DNA probes originating from the HLA class I region were used to detect seven restriction fragment length polymorphisms (RFLPs). Allele frequencies and segregation of each RFLP was determined. Analysis of RFLPs in 38 unrelated homozygotes with hemochromatosis revealed differences in allele frequencies between the control and the hemochromatotic groups but these differences did not reach statistical significance. Some differences persisted, however, even when only controls with the A3 antigen were compared with A3 hemochromatotics. Since both control and hemochromatotic groups were small, further studies will be necessary to ascertain whether these RFLPs could serve to locate the gene responsible for hereditary hemochromatosis. PMID- 2572403 TI - Drug treatment and formation of eicosanoids in patients with chronic inflammatory bowel disease. PMID- 2572404 TI - [Splenic artery aneurysm in cystic media necrosis (Erdheim-Gsell). Spleen-saving surgical therapy]. PMID- 2572405 TI - [Assay of NK cell activity and IL-2 and IFNr production in vitro in patients with epidemic hemorrhagic fever]. AB - In order to study the immune function and pathogenesis of epidemic hemorrhagic fever (EHF), we assayed NK cell cytotoxicity against K562 targets in 95 patients with EHF and the production of IL-2 and IFNr in peripheral blood mononuclear cells (PBMC) after PHA stimulation In contrast to that in normal controls. NK cell activity elevated markedly during febrile phase and reached a highest level during shock and oliguric phase and then gradually decreased to normal. NK cell activity in patients with serious EHF elevated more markedly than that in patients with mild EHF. The production of IL-2 and IFNr in PBMC of patients with EHF after PHA stimulation was normal. The production of IL-2 had significantly positive correlation with NK, cell activity during oliguric, polyuric and convalescent phases, so was the production of IFNr with shock, oliguric, polyuric, and convalescent phases. These results suggests that patients with EHF have still normal cellular immune response. EHF virus may activate NK cells directly and NK cells may participate in the pathogenesis of EHF. PMID- 2572406 TI - [Using urinary diagnostic indices to evaluate acute renal failure in epidemic hemorrhagic fever]. AB - The urinary diagnostic indices were used to evaluate acute renal failure in 118 cases with epidemic hemorrhagic fever (EHF). When the renal failure was mainly caused by acute tubular necrosis, it would occur earlier and persist longer. The authors suggested that the indices are helpful in documenting the severity and identifying the characteristics of acute renal failure in EHF. The mechanisms for developing acute renal failure in EHF was discussed. PMID- 2572407 TI - Effects of biogenic amines and adrenergic drugs on oviposition in the cattle tick Boophilus: evidence for octopaminergic innervation of the oviduct. AB - Oviposition in the cattle tick Boophilus microplus was arrested by octopamine and the alpha-adrenergic agonists clonidine, tolazoline, and naphazoline, at doses of 25 micrograms per tick. The same effect was obtained with a sublethal dose of Amitraz of 10 micrograms per tick. In contrast, the alpha-adrenergic agonists apomorphine, synephrine, phentolamine, metaclopromide and chlorpromazine were ineffective. No mortality was observed at any of the doses tested. Reserpine, which depletes endogenous adrenergic terminals, mimicked the effects of octopamine in blocking oviposition. Acetylcholine agonists also blocked oviposition but dopaminergic agonists did not. Evidence is presented suggesting that control of the oviduct is mediated by octopaminergic receptors. PMID- 2572408 TI - Neurotransmission in the frog retina: possible physiological and histological correlations. AB - In the frog retina, extracellular recordings of transient ganglion cells have shown that the inhibitory surround of the receptive field of these cells was mediated by gamma-aminobutyric acid and acetylcholine (through the nicotinic receptors). Histoautoradiographic and immunocytochemical studies for the two respectively have shown that these neurotransmitters can act through horizontal and amacrine cells. The separation of the ON and OFF channels mediated by glutamate at the bipolar cell level may also be obtained by glycine and/or acetylcholine (through muscarinic receptors). Respective histoautoradiographic and immunocytochemical studies indicate that these neurotransmitters act at the amacrine cell level. These data are consistent with the functional separation of spatial and temporal organization of retinal information, with horizontal cells especially responsible for the spatial organization of the ganglion cell responses and amacrine cells involved in both spatial and temporal organization of the responses. PMID- 2572409 TI - [Use of a chain reaction method of DNA synthesis for analyzing the frequency of restriction polymorphism of the CS-7 DNA locus in the population and in families of patients with mucoviscidosis]. PMID- 2572410 TI - [Therapy of Crohn disease]. PMID- 2572411 TI - Novel patterns of homeotic protein accumulation in the head of the Drosophila embryo. AB - Antibodies that specifically recognize proteins encoded by the homeotic genes: Sex combs reduced, Deformed, labial and proboscipedia, were used to follow the distribution of these gene products during embryogenesis. The position of engrailed-expressing cells was used as a reference and staining conditions were established that could distinguish, among cells expressing engrailed, one of the homeotic proteins or both. Our observations demonstrate two important facts about establishing identity in the head segments. First, in contrast to the overlapping pattern of homeotic gene expression in the trunk segments, we observe a non overlapping pattern in the head for those homeotic proteins required during embryogenesis. In contrast, the spatial accumulation of the protein product of the non-vital proboscipedia locus overlaps partially with the distribution of the Deformed and Sex combs reduced proteins in the maxillary and labial segments, respectively. Second, two of the proteins, Sex combs reduced and Deformed, have different dorsal and ventral patterns of accumulation. Dorsally, these proteins are expressed in segmental domains while, within the ventral region, a parasegmental register is observed. The boundary where this change in pattern occurs coincides with the junction between the ventral neurogenic region and the dorsal epidermis. After contraction of the germ band, when the nerve cord has completely separated from the epidermis, the parasegmental pattern is observed only within the ventral nerve cord while a segmental register is maintained throughout the epidermis. PMID- 2572412 TI - Beta-blockers in the secondary prevention of gastrointestinal haemorrhage in well compensated cirrhotics. A multicentre randomised controlled study. AB - To assess the efficacy of beta-blockers in the prevention of rebleeding in selected cirrhotics and to compare the tolerability, safety of, and patient compliance with, a selective and a non-selective beta-blocker, 94 patients were randomly assigned to propranolol (32), atenolol (32), or placebo (30). Randomisation was made at least 15 days after the bleeding episode. Propranolol was given orally in increasing doses until the resting pulse rate was reduced by approximately 25%. Atenolol was given at a fixed dose of 100 mg/day. Patients were followed up for a mean of 357 days. Rebleeding occurred in 14 patients in the placebo group, 10 in the atenolol group and 8 in the propranolol group. The incidence of rebleeding was significantly lower in patients receiving propranolol than in those on placebo (PR vs PL: p less than 0.01, log-rank test). Atenolol was less effective than propranolol (AT vs PL: p = 0.065, log-rank test) but bleeding-free survival was improved for patients on active drugs compared with those patients on placebo (PR vs PL: p = 0.01; AT vs PL: p = 0.05, log-rank test). Retrospective analysis revealed that, whatever the type of treatment, abstinence from alcohol was crucial in preventing rebleeding. It was concluded that beta-blocker treatment is effective in preventing rebleeding from oesophageal varices in carefully selected alcoholic cirrhotics who survive at least 2 weeks after acute variceal haemorrhage and who cease drinking. PMID- 2572413 TI - Beta-blockers in portal hypertension. An overview. AB - This paper describes a meta-analysis of data from the 5 primary and 15 secondary prevention studies in patients with portal hypertension, in which beta-blockers were compared with inactive control. These studies involved 1525 patients. Overall, the number of patients who suffered bleeding events was 38% lower in the groups treated with beta-blockers (p less than 0.001). The percentage reductions in bleeding-related deaths and in total mortality were 35% and 18%, respectively (p = 0.007 and p = 0.024). More detailed analysis supports the conclusion that propranolol, the beta-blocker predominantly used in these studies, is effective in both the primary and secondary prevention of bleeding events and mortality due to portal hypertension. PMID- 2572414 TI - [Interactions of GABAergic and catecholaminergic neurotransmissions. Effects of dopaminergic and noradrenergic agonists and antagonists on GABA turnover]. AB - The effects of specific D1 and D2 agonists and antagonists on GABA turnover in four brain structures have been studied. GABA turnover was estimated by measuring the accumulation of GABA after GABA-T inhibition with gabaculine. Stimulation of DA receptors by apomorphine, a mixed D1 and D2 agonist or by (+/-)2-(N phenylethyl-N-propyl)amino-5-hydroxytetraline, a selective agonist of D2 receptors, dose-dependently reduced GABA turnover. Both agonists had no effect on GABA levels. S(-)sulpiride, a selective D2 antagonist, had no effect on either GABA levels or GABA turnover. However, sulpiride antagonized the reduction of GABA turnover produced by apomorphine or (+/-)2-(N-phenylethyl-N-propyl)amino-5 hydroxytetraline. By contrast, SKF 38393, a selective D1 agonist, did not appear to influence GABA-mediated inhibitory neurotransmission. SCH 23390, a D1 antagonist, which by itself had no effect on GABA levels and only slightly decreased GABA turnover, did not antagonize the effect of apomorphine. On the contrary, SCH 23390, slightly, but significantly increased the reduction in GABA turnover produced by apomorphine. Furthermore, idaxozan, an alpha 2-antagonist, antagonized the reduction of GABA turnover produced by the alpha 2-agonist clonidine, but did not prevent the effect of apomorphine on GABA turnover. Thus, the tonic inhibition exerted by DA on GABA-mediated neurotransmission seems to be mainly controlled by D2 receptors. PMID- 2572415 TI - Nucleolar inhibition in prolactin cells after combined thyroid hormone plus lisuride treatment in estrogenized rats. AB - The growth reaction of the rat adenohypophysis to estradiol was previously found to be potentiated by dopaminergic antagonists (perphenazine) and inhibited by thyroid hormones and dopaminergic agonists. In the present experiment a combination of estradiol, lisuride and thyroid hormones was tested. Sixty male Wistar rats were divided into 6 groups: 1. controls; 2. estradiol benozate (E) in aqueous microcrystal suspension 1 mg/rat twice a week; 3. Thyreoidin (SPOFA) 0.2% in the food (T); E + T; 5. E + lisuride (N-D-6-methyl-8-isoergonelyl)-N'N-diethyl carbamide hydrogen maleate, 200 micrograms/rat per day in the food (L); 5. E + T + L. After 3 weeks the adenohypophysial weights, histology (PAS Orange G, indirect immunoperoxidase technique) and electron microscopic morphometry were examined. The increase in adenohypophysial weight was inhibited by T, L and significantly more by the combination T + L. The increased incidence of prolactin cells after estradiol was inhibited by T and L and equally by the combination T + L. The effect of both inhibitory substances (T + L) was also additive on the relative size of the nucleoli of PRL cells: marked nucleolar inhibition similar to that produced by cytostatics was observed in the E + T + L group of animals. PMID- 2572416 TI - Electrophoretic behavior and partial characterization of disease-associated serum forms of gamma-glutamyltransferase. AB - We have recently devised an improved procedure for the rapid electrophoretic separation of multiple forms of serum gamma-glutamyltransferase (GGT). This procedure is based on the separation on cellulose acetate strips, usually employed for lipoprotein electrophoresis, followed by visualization with a fluorescent reagent. The method is highly sensitive and the fractions are more clearly resolved than with other procedures. Reference intervals have been evaluated in the sera from 142 healthy subjects and the patterns (two GGT forms comigrating with alpha 1 and alpha 2-globulin) are reproducible. In 150 sera from patients with various hepatobiliary diseases (including neoplasias), acute pancreatitis and non liver-involving neoplasias, we observed some disease specific GGT forms: an albumin comigrating enzyme (Alb-GGT) specific of liver neoplasia; a gamma-globulin comigrating GGT (gamma-GGT) and a nonmigrating isoform (dep-GGT) both specifically associated to extrahepatic jaundice. Multiple lipoprotein fraction precipitation showed that beta-, gamma- and dep-GGT are complexes between GGT and low density lipoprotein and very low density lipoproteins (LDL + VLDL), and that some of the alpha 1-GGT from cirrhotic patients is a complex between GGT and high density lipoprotein (HDL). GGT fractions from normal subjects and Alb-GGT from patients with liver neoplasia do not appear to be complexed with lipoproteins.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572417 TI - Hepatocyte heterogeneity in glutamate metabolism and bidirectional transport in perfused rat liver. AB - 1. The metabolic fate of infused [1-14C]glutamate was studied in perfused rat liver. The 14C label taken up by the liver was recovered to 85 +/- 2% as 14CO2 and [14C]glutamine. Whereas 14CO2 production accounted for about 70% of the [1 14C]glutamate taken up under conditions of low endogenous rates of glutamine synthesis, stepwise stimulation of glutamine synthesis by NH4Cl increased 14C incorporation into glutamine at the expense of 14CO2 production. Extrapolation to maximal rates of hepatic glutamine synthesis yielded an about 100% utilization of vascular glutamate taken up by the liver for glutamine synthesis. This was observed in both, antegrade and retrograde perfusions and suggests an almost exclusive uptake of glutamate into perivenous glutamine-synthetase-containing hepatocytes. 2. Glutamate was simultaneously taken up and released from perfused rat liver. At a near-physiological influent glutamate concentration (0.1 mM), the rates of unidirectional glutamate influx and efflux were similar (about 100 and 120 nmol g-1 min-1, respectively). 3. During infusion of [1-14C]oxoglutarate (50 microM), addition of glutamate (2 mM) did not affect hepatic uptake of [1 14C]oxoglutarate. However, it increased labeled glutamate release from the liver about 10-fold (from 9 +/- 2 to 86 +/- 20 nmol g-1 min-1; n = 4), whereas 14CO2 production from labeled oxoglutarate decreased by about 40%. This suggests not only different mechanisms of oxoglutarate and glutamate transport across the plasma membrane, but also points to a glutamate/glutamate exchange. 4. Oxoglutarate was recently shown to be taken up almost exclusively by perivenous glutamine-synthetase-containing hepatocytes [Stoll, B & Haussinger, D. (1989) Eur. J. Biochem. 181, 709-716] and [1-14C]oxoglutarate (9 microM) was used to label selectively the intracellular glutamate pool in this perivenous cell population. The specific radioactivity of this intracellular (perivenous) glutamate pool was assessed by measuring the specific radioactivity of newly synthesized glutamine which is continuously released from these cells into the perfusate. Comparison of the specific radioactivities of glutamine and glutamate released from perivenous cells indicates that about 60% of total glutamate release from the liver is derived from the perivenous glutamine-synthetase containing cell population. Following addition of unlabeled glutamate (0.1 mM), unidirectional glutamate efflux from perivenous cells increased from about 30 to 80 nmol g-1 min-1, whereas glutamate efflux from non-perivenous (presumably periportal) hepatocytes remained largely unaltered (i.e. 20-30 nmol g-1 min-1). 5. It is concluded that, in the intact liver, vascular glutamate is almost exclusively taken up by the small perivenous hepatocyte population containing glutamine synthetase. PMID- 2572418 TI - Inhibition of transmitter release by botulinum neurotoxin A. Contribution of various fragments to the intoxication process. AB - 1. The contribution of a proteolytic fragment (H2L) of botulinum neurotoxin type A (comprised of the aminoterminal region of the heavy-chain disulphide-linked to the light chain) to inhibition of neurotransmitter release was investigated, using central cholinergic synapses of Aplysia, rodent nerve-diaphragm preparations and cerebrocortical synaptosomes. 2. No reduction in neurotransmitter release was observed following external application to these preparations of highly purified H2L or after intracellular injection into Aplysia neurons. 3. The lack of activity was not the result of alteration in the light chain of H2L during preparation of the latter because (a) renaturation of this light chain with intact heavy chain produced a toxic di-chain form and (b) simultaneous application of heavy chain and light chain from H2L inhibited transmitter release in Aplysia. 4. Bath application of H2L and heavy chain together inhibited release of transmitter; however, at the neuromuscular junction the potency of this mixture was much lower than that of native toxin. A similar blockade resulted when heavy chain was applied intracellularly and H2L added to the bath, demonstrating that H2L is taken up into cholinergic neurons of Aplysia. This uptake is shown to be mediated by the amino-terminal moiety of heavy chain (H2), because bath application of light chain plus H2 led to a decrease in acetylcholine release from a neuron that had been injected with heavy chain. 5. A role within the neuron is implicated for a carboxy-terminal portion of heavy chain (H1) since intracellular injection of light chain and H2 did not affect transmitter release. Although the situation is unclear in mammalian nerves, these collective findings indicate that blockade of transmitter release in Aplysia neurons requires the intracellular presence of light chain and H1 (by inference), whilst H2 contributes to the internalization step. PMID- 2572419 TI - A comparison of the effects of adrenaline and noradrenaline on human heart: the role of beta 1- and beta 2-adrenoceptors in the stimulation of adenylate cyclase and contractile force. AB - The stimulant effects of adrenaline and noradrenaline on contractile force and adenylate cyclase, mediated through beta 1 and beta 2-adrenoceptors, are analysed in isolated atrial and ventricular myocardium of man. The tissues were obtained from patients without advanced heart failure undergoing heart surgery. Usually, both adrenaline and noradrenaline stimulated adenylate cyclase predominantly through ventricular and atrial beta 2-adrenoceptors. Because the relative density of beta 2-adrenoceptors is usually smaller than that of beta 1-adrenoceptors, stimulation of one beta 2-adrenoceptor leads to the production of up to 10 times more cyclic AMP molecules than does stimulation of one beta 1-adrenoceptor. Adrenaline and noradrenaline maximally enhance contractile force through both atrial and ventricular beta 1-adrenoceptors. Adrenaline can also maximally enhance contractile force through atrial beta 2-adrenoceptors. In the ventricle, adrenaline increases force via beta 2-adrenoceptors by up to 60% of its maximal beta 1 response. Noradrenaline can increase atrial and ventricular contractile force through beta 2-adrenoceptors but only at high concentrations. Unexpectedly, in atria from patients treated with the beta 1-selective antagonist atenolol, contractile responses to adrenaline are markedly and selectively augmented through activation of beta 2-adrenoceptors. In atria from atenolol-treated patients equi-inotropic concentrations of adrenaline and noradrenaline acting through beta 2 and beta 1-adrenoceptors, respectively, cause similar increases of cyclic AMP and of cyclic AMP-dependent protein kinase activity. PMID- 2572421 TI - Central and peripheral adrenergic receptor agonists in heart failure. AB - The subtypes of alpha- and beta- and dopaminergic receptors have been identified along with specific agonists and antagonists. The effects of alpha-receptor agonists on haemodynamics results from the interaction of relative changes in vascular tone and increase in myocardial contractility. Predominantly, beta 1 agonists improve cardiac performance by increasing contractility which is usually accompanied by increased myocardial oxygen consumption. Dopaminergic receptor agonists improve left ventricular function primarily by reduction in systemic vascular resistance usually without a change in myocardial oxygen consumption. beta 2-receptor agonists improve cardiac function both by increasing contractility and by peripheral vasodilation usually without an increase in metabolic cost. The effects of various adrenoreceptor agonists such as dobutamine, dopamine, prenalterol, levodopa and dopexamine on systemic and coronary haemodynamics and on myocardial energetics in heart failure are reviewed. PMID- 2572420 TI - The beta-adrenergic receptor-adenylate cyclase complex as a target for therapeutic intervention in heart failure. AB - In the human heart the beta-adrenergic pathways are the primary means of increasing cardiac performance in response to acute or chronic stress. Control of beta pathway function is achieved by changes in the receptors themselves, and to a lesser degree by adjustments in the inhibitory G protein (Gi). In heart failure myocardial beta-receptor function is substantially reduced, but the beta-receptor pathways are so powerful that they remain capable of supporting inotropic function. Certain therapeutic interventions that improve exercise performance in heart failure can partially restore beta-receptor pathway function to normal; these interventions include beta-blocker therapy and treatment with angiotensin converting enzyme inhibitors. Other types of therapy, such as chronic administration of beta-adrenergic agonists, may produce undesirable effects by increasing beta-receptor subsensitivity; however, the effects of beta-agonists on beta-receptor function are somewhat unpredictable and certain beta-agonists do not appear to produce much desensitization. Finally, the failing human heart is in effect partially denervated due to depletion of neuronal norepinephrine, and consequently in advanced heart failure beta-agonists that possess an indirect component of action will be less effective than exclusively direct-acting agents. These observations indicate that the baseline status and the intervention associated behaviour of the human myocardial beta-adrenergic receptor systems need to be considered in developing therapeutic strategies in congestive heart failure. PMID- 2572422 TI - Characteristics, prognosis and treatment of the ventricular arrhythmias of right ventricular dysplasia. AB - We studied 58 cases of arrhythmogenic right ventricular dysplasia (ARVD). Sustained monomorphic ventricular tachycardia (VT) was present in 50 patients, ventricular fibrillation (VF) in three (two also having VT), and non-sustained VT in the remaining seven. Different morphologies of VT were documented in 24 of the 50 patients with sustained VT. They had a left bundle branch block pattern in 96% of cases, without extreme deviation of the QRS axis, and a QRS relatively narrow and ample. These sustained VTs were triggered by provocative techniques. Holter recordings showed frequent ventricular extrasystoles in the great majority of cases. They were polymorphic in 78%, with runs of VT in 59% of patients. Spontaneous onset of VT occurred during exercise in 60% of cases, preceded by a sinus rate increase when recording was available. This is more frequent in angiographically localized forms of ARVD than in diffuse forms, and tends to disappear during follow-up. Only four cardiac deaths occurred after a follow-up of 8.8 +/- 7.2 years: three by acute heart failure, and only one by recurrent VF. Spontaneous disappearance of VT which became non-inducible was seen in four cases. Single antiarrhythmic drug therapy was judged satisfactory in 21 cases, and combined therapy in 19 other cases. Surgery of fulguration was performed in 17 cases, with 14 successes (nine of them with combined antiarrhythmic therapy). Despite a frequent lack of control of VT by antiarrhythmic drugs, the follow-up of ARVD seems good in patients with sustained VT. Some arguments favour the concept of a diffuse and progressive disease. PMID- 2572423 TI - XbaI restriction fragment length polymorphism of apolipoprotein B in Swedish myocardial infarction patients. AB - Apolipoprotein B (apoB) is a major importance to the metabolism of lipoproteins, and there is also evidence which suggests that apoB plays a central role in atherogenesis. In order to study whether there is a link between one of the mutations of the apoB gene and premature coronary heart disease, the frequency of the XbaI RFLP for the apoB gene was analysed in 52 male myocardial infarction patients. These were compared with a control group matched for age and sex (n = 52), and a random population sample of middle-aged men (n = 106). Two alleles were identified by the presence (X2) or the absence (X1) of an XbaI cleavage site. A somewhat higher frequency of the X2 allele was seen among the patients, however there was no significant difference between patients and controls regarding the genotypes or allele frequencies. This observation does not confirm one earlier report where a higher frequency of the X1 allele was seen in myocardial infarction patients. Differences between the studied populations or epidemiological designs of the studies might explain the diverging results. Further studies are evidently needed to fully resolve the relation between the XbaI RFLP and risk of atherosclerotic disease or lipoprotein metabolism. PMID- 2572425 TI - Dihydrexidine, a novel selective high potency full dopamine D-1 receptor agonist. PMID- 2572424 TI - The enantiomers of the dopamine agonist N-0437: in vivo and in vitro effects on the release of striatal dopamine. AB - The enantiomers of the potent and selective dopamine (DA) D-2 receptor agonist 2 (N-propyl-N-2-thienylethyl-amino)-5-hydroxytetralin, N-0437, were tested for their pharmacological actions on DA D-2 autoreceptors in vivo, by measuring DA release by microdialysis during local administration of both drugs and in vitro, by measuring their effects on the electrically stimulated release of [3H]DA from striatal slices. In both experimental situations (-)-N-0437, at low doses, acted as an agonist on receptors controlling DA release. However, in vivo at a concentration of 10 microM (-)-N-0437 induced a short-lasting increase in DA release and in vitro the inhibitory effect of (-)-N-0437 was significantly less pronounced at higher concentrations (1-10 microM). (+)-N-0437 (0.1-10 microM) showed an antagonistic action both in vivo and in vitro. Following inhibition of the neuronal impulse flow with tetrodotoxin, (+)-N-0437 failed to increase DA release suggesting the effect of this enantiomer is not associated with an amphetamine-like action on the nerve terminal. The use of the DA re-uptake inhibitor GBR 12909 confirmed the antagonism of (+)-N-0437 towards DA receptors. Since it is known that (+)-N-0437 acts as an agonist on DA D-2 autoreceptors controlling the synthesis of DA, these results provide additional evidence for the existence of distinct DA D-2 autoreceptor populations involved in the release and synthesis of DA. The differential actions of (-)- and (+)-N-0437 gave rise to mutual antagonism of the actions of the enantiomers both in vivo and in vitro, thus providing a strong argument for using the enantiomers instead of the racemate in clinical situations. PMID- 2572426 TI - Stereoselective reversal of MPTP-induced parkinsonism in the marmoset after dermal application of N-0437. AB - The selective dopamine D-2 receptor agonist N-0437 produced a rapid and dose dependent reversal of motor deficits lasting 90-120 min following i.p. or oral administration of the racemate to MPTP-treated common marmosets. In contrast, topical application of (+/-)-, (+)- or (-)-N-0437 to the skin of MPTP-treated animals did not alter locomotor activity in the initial 4 h although other motor disabilities were reduced. However, 24 h following application of the racemate or the (-) enantiomer both locomotor activity and the other motor deficits induced by MPTP were improved. The increase in locomotor activity returned to basal values by 48-52 h following application of the racemate to the skin and by 72-76 h following administration of (-)-N-0437; the other motor deficits induced by MPTP were reduced for up to 72-76 h by both (+/-)- and (-)-N-0437. Application to skin of the (+) enantiomer produced no behavioural improvement or stimulation of locomotor activity. Transdermal administration of the active enantiomer of N-0437 may be of value in producing a prolonged reversal of parkinsonian motor deficits in man. PMID- 2572427 TI - Attenuation of hypothermic effects of ethanol by alpha 2-adrenoceptor blockers. AB - Effects of selective alpha 2-adrenoceptor antagonists, atipamezole and idazoxan, on ethanol-induced hypothermia were investigated in mice. Ethanol significantly reduced (P less than 0.001) core temperature whilst both alpha 2-adrenoceptor antagonists were without effect when administered alone. However, both the 1 and 3 mg/kg doses of atipamezole significantly (P less than 0.05) attenuated the ethanol-induced reduction in body temperature 20 and 40 min after administration. The 3 mg/kg dose of idazoxan (but not the 1 mg/kg dose) also significantly (P less than 0.05) attenuated ethanol's hypothermic effect 20 min after administration but this effect was not statistically significant at 40 min. In a subsequent experiment using lower doses of atipamezole (0.03-1.0 mg/kg) the attenuation of ethanol-induced hypothermia caused by atipamezole was found to be dose-related. The effect of the benzodiazepine inverse agonist Ro 15-4513 on ethanol-induced hypothermia was also investigated. This compound possessed an intrinsic hypothermic action but neither attenuated nor enhanced the hypothermic effect of ethanol. These results suggest that alpha 2-adrenoceptor can, at least partially, modulate the hypothermic effects of ethanol. PMID- 2572428 TI - HA-966 acts at a modulatory glycine site to inhibit N-methyl-D-aspartate-evoked neurotransmitter release. AB - The role of endogenous glycine in supporting N-methyl-D-aspartate (NMDA)-evoked neurotransmitter release was investigated. HA-966 (1-hydroxy-3-aminopyrrolidone 2) inhibited NMDA-evoked release of [3H]norepinephrine from rat hippocampal brain slices, but was much less effective in inhibiting [3H]norepinephrine release evoked by kainic acid (KA). Glycine (1 mM) reversed the HA-966 (1 mM) antagonism of NMDA-evoked release of [3H]norepinephrine. Strychnine (10 microM) had no effect on the ability of glycine to reverse HA-966 antagonism of NMDA-evoked neurotransmitter release. Other amino acids were also capable of reversing the HA 966 antagonism of NMDA-evoked [3H]norepinephrine release with a rank order of potency: D-serine greater than or equal to glycine much greater than L-serine approximately beta-alanine. These same compounds inhibited strychnine-insensitive [3H]glycine binding to rat cortical membrane fragments with a rank order of potency: glycine greater than D-serine much greater than L-serine greater than or equal to beta-alanine. In addition, HA-966 inhibited [3H]glycine binding (IC50 = 8.5 microM). The results suggest that HA-966 antagonism of NMDA-evoked neurotransmitter release is due to the inhibition of endogenous glycine acting at a strychnine-insensitive modulatory glycine site associated with the NMDA receptor/ionophore complex. PMID- 2572429 TI - Antipsychotic substances and dopamine in the rat brain; behavioral studies reveal distinct dopamine receptor systems. AB - Graded doses of apomorphine were injected into terminal areas of the nigrostriatal, mesolimbic and mesocortical dopamine (DA) systems of the rat brain. Injection of high doses of apomorphine into the nucleus caudatus elicited stereotyped sniffing, but did not affect the motility of the rat. Low and high doses of apomorphine injected into the nucleus accumbens decreased and increased motility, respectively, without changing sniffing behavior. Injection of both low and high doses of apomorphine into the pyriform cortex increased sniffing behavior, but did not affect motility. All these apomorphine-induced behavioral responses were antagonized by local pretreatment with the typical neuroleptic, haloperidol, and the atypical neuroleptic, sulpiride, albeit with different potencies as revealed from the calculated ED50 values (ranging from 0.18-20,462 fmol). Local pretreatment with the antipsychotic peptide, des-enkephalin-gamma endorphin (DE gamma E), antagonized the behavioral changes induced by injecting low and high doses of apomorphine into the pyriform cortex and low doses into the nucleus accumbens (ED50: 0.22-20.6 fmol), but did not affect the behavioral effects elicited by injecting high doses of apomorphine into the nucleus caudatus or the nucleus accumbens. It is proposed that two distinct types of DA receptor systems are present in the rat brain. These are characterized by the slope of the dose-response curve for the substances to antagonize the apomorphine-induced behavioral effects and by the effectiveness of DE gamma E in this respect. The antipsychotic effects of these compounds may be mediated by DA systems in the nucleus accumbens or in the pyriform cortex; this last system is especially sensitive to the three antipsychotic substances and is equally affected by them (ED50 value for the antagonism of the apomorphine-induced effects: 0.18-0.71 fmol). PMID- 2572430 TI - Effects of prolactin on tyrosine hydroxylase activity of central dopaminergic neurons of male rats. AB - Previous work from our laboratory has indicated that ovine prolactin can produce a rapid stimulation (within 1 h) of dopamine release from nigrostriatal and mesolimbic dopaminergic neurons of male rats. In the present experiment we attempted to determine whether this effect of prolactin was a function of an increase in the activation of dopamine synthesis. To examine this possibility we used the drug NSD 1015. This drug is a decarboxylase inhibitor that leads to dopa accumulation and is used as an index of in vivo tyrosine hydroxylase activity. Our results demonstrated that ovine prolactin increased turnover of dopamine but had no effect upon tyrosine hydroxylase activation (up to 4 h) in both dorsal and ventral striatal dopaminergic neurons. In contrast, ovine prolacting had a clear effect (within 4 h) on tyrosine hydroxylase activity in mediobasal hypothalamic dopaminergic neurons. These results suggest that ovine prolactin can differentially alter synthesis/release processes of dopamine from these sites. PMID- 2572431 TI - Stannous chloride-induced increase in calcium entry into motor nerve terminals of the frog. AB - The effects of SnCl2 on potentials recorded extracellularly from motor nerve terminals of the bullfrog were studied to elucidate the mechanism of the SnCl2 induced facilitation of evoked transmitter release. Under conditions in which the muscle preparations were pretreated with d-tubocurarine and tetraethylammonium in a K+-free medium, SnCl2 (50 microM) augmented the prolonged positive deflection ascribed to the inward Ca2+ current, an effect which was reduced by addition of Cd2+. The results suggest that SnCl2 could increase Ca2+ entry into the nerve terminals. PMID- 2572432 TI - Differential effect of typical and atypical antipsychotic drugs on the suppressant action of 2-methylserotonin on medical prefrontal cortical cells: a microiontophoretic study. PMID- 2572433 TI - Two phases of contractile response in rat isolated vas deferens and their regulation by adenosine and alpha-receptors. AB - Contractions to transmural electrical stimulation and exogenous norepinephrine were recorded in isolated longitudinal segments of rat vas deferens. Electrical stimulation for 30 s produced a biphasic contraction in the vas deferens consisting of a rapid, transient response (Phase I), followed by a slowly developing, sustained contraction (Phase II). N6-Cyclohexyladenosine (CHA), a selective adenosine1 (A1)-receptor agonist, attenuated in a concentration dependent manner the Phase I contractile response, while having little effect on the Phase II response. In contrast, 2-(phenylamino)adenosine (CV-1808), a selective adenosine2 (A2)-receptor agonist had little effect on either contractile phase. CHA did not inhibit the contraction to exogenous norepinephrine, suggesting that A1-receptors were located at a presynaptic site. The relatively selective alpha 2-receptor agonist clonidine produced the same pattern of contractile inhibition as CHA. The inhibitory effect of CHA on the Phase I contractile response in the vas deferens could be antagonized by the selective A1-receptor antagonist 8-cyclopentyltheophylline, while the selective alpha 2-receptor antagonist idazoxan preferentially antagonized the inhibitory effect of clonidine on the Phase I response. Both the Phase I and Phase II contractile responses were reduced by the selective alpha 1-adrenoceptor antagonist prazosin and the ATP analog alpha, beta-methylene adenosine triphosphate (alpha, beta-methylene ATP), suggesting that norepinephrine and ATP are coreleased as neurotransmitters for both responses. The results of the present study demonstrate that in the rat vas deferens the presynaptic inhibitory effects of adenosine is mediated by the A1-receptor subtype, and that both A1- and alpha 2-receptor agonists exert a selective inhibitory effect on the Phase I contractile response to electrical stimulation. PMID- 2572434 TI - Behavioral differentiation of benzodiazepine ligands after repeated administration in baboons. AB - Baboons received repeated daily administration of saline, 5.6 mg/kg midazolam, 5.0 mg/kg flumazenil (Ro15-1788), 3.2 mg/kg 3-carboethoxy-beta-carboline hydrochloride (beta CCE) or 10 mg/kg beta CCE for 5 days. Behavioral signs of sedation and excitation were scored for 1 h after i.m. injections. Daily administration of these benzodiazepine-receptor ligands differentiated their behavioral effects; repeated midazolam resulted in tolerance to the sedative and ataxic effects; repeated beta CCE resulted in sensitization to its convulsant properties; and repeated flumazenil or saline produced no changes in behavior. In a second study, baboons received repeated injections of midazolam (5.6, 11.2 or 20 mg/kg per day) for 6 days. All three groups became tolerant to the sedative and ataxic effects of midazolam. Acute injections of flumazenil (5.0 mg/kg) on day 5 produced a dose-dependent withdrawal syndrome. This flumazenil treatment produced a slight attenuation in the degree of tolerance to midazolam on day 6, suggesting that receptor sensitivity to the benzodiazepine agonist may have partially reset. PMID- 2572435 TI - MDMA (3,4-methylenedioxymethamphetamine) inhibits the firing of dorsal raphe neurons in brain slices via release of serotonin. AB - The effects of MDMA (3,4-methylenedioxymethamphetamine) on the activity of serotonin (5-HT)-containing dorsal raphe neurons were characterized using extracellular single-unit recording and microdialysis techniques in the in vitro midbrain slice preparation. Addition of (+)-MDMA, (-)-MDMA or p-chloroamphetamine (PCA) to the superfusate (final concentration 3-100 microM) produced a concentration-dependent inhibition of 5-HT cell firing which was reversible and reproducible. Based upon IC50 values, (+)-MDMA was 2- to 3-fold more potent than (-)-MDMA. Pretreatment with the selective 5-HT uptake inhibitor fluoxetine, at a concentration which had no effect on baseline firing (20 microM), blocked the inhibitory effect of (+)-MDMA and PCA on dorsal raphe neurons. The selective norepinephrine uptake inhibitor desipramine (20 microM) was ineffective. In a parallel series of experiments, microdialysis probes resting on the brain slice surface provided a means to estimate 5-HT release from the dorsal raphe nucleus. (+)-MDMA (100 microM) caused the release of measureable quantities of 5-HT with a time course which corresponded to the change in dorsal raphe cell firing rate. Taken together, these data suggest that MDMA acts indirectly to inhibit dorsal raphe neurons through release of endogenous 5-HT. PMID- 2572436 TI - Beta-adrenoceptors in human brain labelled with [3H]dihydroalprenolol and [3H]CGP 12177. AB - beta-Adrenoceptor binding sites were characterised and quantitated in post-mortem human brain with [3H]dihydroalprenolol ([3H]DHA) and [3H]CGP 12177. In cerebral cortex, isoprenaline and propranolol displaced both radioligands with uniform affinity. Practolol and CGP 20712A (selective beta 1-adrenoceptor antagonists) displaced with high affinity from a greater proportion of sites than ICI 118,551 and IPS 339 (selective beta 2-adrenoceptor antagonists). In cerebellum, propranolol displaced both radioligands with uniform affinity. ICI 118,551 displaced with high affinity from a greater proportion of sites than CGP 20712A. The density of total beta-adrenoceptors (defined with isoprenaline) and of beta 1 and beta 2-adrenoceptors (defined with CGP 20712A and ICI 118,551 respectively) was studied by saturation binding of both radioligands in 13 brain areas. beta Adrenoceptor density was higher in caudate, putamen and nucleus accumbens (100 120 fmol/mg protein) than cortex (50-70 fmol/mg protein) and densities were lowest in hypothalamus and cerebellum (27-38 fmol/mg protein). The proportion of beta 1-adrenoceptors (as a % of total beta-adrenoceptors) was high in caudate (80%), putamen (80%) and cortex (60-70%) and lower in hippocampus (40%) and cerebellum (30%). Both radioligands labelled a very similar number of beta adrenoceptors in all brain regions studied. PMID- 2572438 TI - Demonstration of alpha 1A- and alpha 1B-adrenoceptor binding sites in human brain tissue. AB - Radioligand binding studies suggest that alpha 1-adrenoceptor recognition sites are heterogeneous. Several adrenergic agents discriminate between two adrenoceptor binding sites designated alpha 1A and alpha 1B. In the present study we demonstrate for the first time that these two subtypes exist in the human brain. 5-Methyl-urapidil and (+)-niguldipine, which have previously been shown to be alpha 1A-selective, inhibited [3H]prazosin binding to cortical membranes in a biphasic manner. The irreversible alpha 1B-ligand, chloroethylclonidine, preferentially eliminated the binding sites with low affinity for (+) niguldipine. In contrast, BE 2254 and unlabelled prazosin displaced the radioligand in a monophasic manner. The IC50 values for prazosin were not affected by pretreatment of the membranes with chloroethylclonidine. Our data on human brain membranes are in excellent agreement with recent findings in rat tissues and suggest that the alpha 1-adrenoceptor subtypes in human brain are similar to those in rat tissues. PMID- 2572439 TI - [Review: experimental studies on Creutzfeldt-Jakob disease]. PMID- 2572437 TI - Effects of sulfasalazine and a sulfasalazine analogue on the formation of lipoxygenase and cyclooxygenase products. AB - A sulfasalazine analogue, 5'-(2,4-dichlorobenzoyl)2'-hydroxyphenylacetic acid (CL 42A), potently inhibited the formation of 5-lipoxygenase products (leukotrienes B4 and C4 and 5-hydroxyeicosatetraenoic acid) by human leukocytes. Half-maximal inhibition of leukotriene production was obtained with 5 and 10 microM CL 42A after stimulation with serum-treated zymosan or ionophore A23187, respectively. CL 42A was equipotent to nordihydroguaiaretic acid and about 50 times more potent than sulfasalazine and benoxaprofen in studies on the inhibition of LTB4 formation in leukocyte suspensions stimulated with serum-treated zymosan. Furthermore, CL 42A had no inhibitory effect on the production of 15 hydroxyeicosatetraenoic acid after incubation of human leukocytes with ionophore A23187 in the presence of exogenous arachidonic acid. Sulfasalazine inhibited the synthesis of 5-lipoxygenase products (5-hydroxyeicosatetraenoic acid and leukotriene B4: IC50 250 microM, leukotriene C4: IC50 100 microM) in a concentration-dependent manner but had no effect on 15-hydroxyeicosatetraenoic acid formation. The metabolites of sulfasalazine, sulfapyridine and 5 aminosalicylic acid, and the isomer, 4-aminosalicylic acid, were all less potent than sulfasalazine as inhibitors of leukotriene formation. Both CL 42A (IC50 20 microM) and sulfasalazine (IC50 500 microM) inhibited the synthesis of thromboxane B2 and hydroxyheptadecatrienoic acid in human platelet suspensions after arachidonic acid stimulation. However, while CL 42A inhibited cyclooxygenase, the inhibitory effect of sulfasalazine was exerted mainly on thromboxane synthase. The platelet formation of 12-hydroxyeicosatetraenoic acid was not inhibited by CL 42A whereas sulfasalazine had a weak inhibitory effect. PMID- 2572440 TI - Restriction fragment length polymorphism of cardiac myosin heavy chain gene in rats and its strain distribution. AB - The distribution of an RFLP in EcoRI fragments of the cardiac myosin heavy chain gene among 29 strains of laboratory rats was examined. Southern blot hybridization of rat genomic DNAs with rat cardiac myosin heavy chain cDNA as a probe demonstrated an interstrain variation in one of eight EcoRI fragments. Of the 28 inbred strains examined, 10 had a fragment of 10 kbp, whereas 18 had a fragment of 7.5 kbp. The 15 samples of the remaining strain (Iar: WI outbred stock) had fragments of either 7.5 kbp or 10 and 7.5 kbp, indicating that this strain has maintained heterogeneity of these fragments. PMID- 2572441 TI - H-ras gene is expressed at the G1 phase in primary cultures of hepatocytes. AB - The expression of c-H-ras and proliferating cell nuclear antigen (PCNA) in primary cultures of rat hepatocytes was determined in order to elucidate the relationship between the c-H-ras gene and the S phase of the cell cycle. In cells treated with EGF, elevation of c-H-ras expression was detected at the 22nd, 34th, 44th, and 54th h after plating, PCNA expression and DNA synthesis were detected at the 44th and 54th h. In cells without EGF treatment, only c-H-ras expression was detected at the 44th and 54th h. In our previous report, we showed that c-myc expression increased within several hours after plating, suggesting that isolated hepatocytes traverse from G0 to G1 under culture conditions, regardless of EGF treatment. These results clearly showed that the c-H-ras gene of adult rat hepatocytes was expressed in the mid-to-late G1 phase of the cell cycle as well as in the early S phase in primary culture. PMID- 2572442 TI - Translation of poly(A)+ mRNA encoding CFU-S proliferation stimulator of human fetal liver origin in Xenopus laevis oocytes. AB - Using the sodium dodecyl sulfate/chloroform/phenol method, total RNA was extracted from human fetal liver tissue with a yield of 870-1060 micrograms/g of tissue; mRNA was obtained after chromatography on an oligo(dT)-cellulose column. Experimental studies demonstrated the translation of a poly(A)+ mRNA encoding proliferation stimulator of human fetal liver origin in Xenopus laevis oocytes. PMID- 2572443 TI - Blood-derived stem cell collection in acute nonlymphoblastic leukemia: predictive factors for a good yield. AB - Blood-derived stem cell (BDSC) autografting represents an interesting theoretical approach to the treatment of acute nonlymphoblastic leukemia (ANLL). The feasibility and safety of this procedure have now been established by several observations of rapid hematopoietic recovery following high-dose chemotherapy and autologous reinfusion of BDSCs harvested during remission. Using clonogenic assays for granulocyte-macrophage colony-forming units (CFU-GM) and granulocyte erythrocyte monocyte colony-forming units (CFU-GEM), we tested peripheral blood samples from 20 consecutive patients recovering from induction chemotherapy for ANLL. Results were correlated with other clinical and hematological factors in order to define optimal criteria for successful BDSC harvesting. Two different patterns of increment were observed in the number of peripheral blood progenitor cells during early recovery from chemotherapy in our patients, suggesting that a total period of 10-12 days should be covered for ideal BDSC harvesting by cytapheresis. Associated clinical factors that appear predictive for a better yield are: 1) chemotherapy with daunorubicin, cytosine arabinoside, and thioguanine (DAT), 2) synchronous recovery of monocytes and platelets, and 3) a good overall performance status of the patient. Complete remission status and absence of ongoing infection should also be considered when selecting patients for BDSC harvesting. No correlation was found between the number of circulating CFUs, as indicated by the clonogenic assays, and that of either My-10- or HLA-Dr positive cells identified by immunofluorescence on the same blood samples. In this study, less than 50% of patients with newly diagnosed ANLL would have been considered good candidates for a BDSC harvesting program. PMID- 2572444 TI - Impaired reference memory and reduced glutamergic activity in rats with temporo entorhinal connections disrupted. AB - The purpose of the present study was to investigate whether effects of temporo entorhinal disconnections on acquisition and retention of a visual discrimination task might be associated with neurochemical dysfunctions. The results revealed that the present lesions impaired both acquisition and retention of the discrimination task. This impairment was accompanied by decreased glutamergic activity in both temporal and entorhinal cortices. No changes were seen in levels of acetylcholine or GABA. Further, the distribution of glutamate/aspartate was related to both regional and hemispheric differences. The results are discussed in terms of a highly integrative role of the lateral entorhinal cortex and in terms of other putative neurotransmitter systems involved in the function of memory. PMID- 2572445 TI - A role for N-methyl-D-aspartate receptors in norepinephrine-induced long-lasting potentiation in the dentate gyrus. AB - Mechanisms of action of norepinephrine (NE) on dentate gyrus granule cells were studied in rat hippocampal slices using extra- and intracellular recordings and measurements of stimulus and amino acid-induced changes in extracellular Ca2+ and K+ concentration. Bath application of NE (10-50 microM) induced long-lasting potentiation of perforant path evoked potentials, and markedly enhanced high frequency stimulus-induced Ca2+ influx and K+ efflux, actions blocked by beta receptor antagonists and mimicked by beta agonists. Enhanced Ca2+ influx was primarily postsynaptic, since presynaptic delta [Ca2+]o in the stratum moleculare synaptic field was not altered by NE. Interestingly, the potentiation of both ionic fluxes and evoked population potentials were antagonized by the N-methyl-D aspartate (NMDA) receptor antagonist 2-amino-5-phosphonovalerate (APV). Furthermore, NE selectively enhanced the delta [Ca2+]o delta [K+]o and extracellular slow negative field potentials elicited by iontophoretically applied NMDA, but not those induced by the excitatory amino acid quisqualate. These results suggest that granule cell influx of Ca2+ through NMDA ionophores is enhanced by NE via beta-receptor activation. In intracellular recordings, NE depolarized granule cells (4.8 +/- 1.1 mV), and increased input resistance (RN) by 34 +/- 6.5%. These actions were also blocked by either the beta-antagonist propranolol or specific beta 1-blocker metoprolol. Moreover, the depolarization and RN increase persisted for long periods (93 +/- 12 min) after NE washout. In contrast, while NE, in the presence of APV, still depolarized granule cells and increased RN, APV made these actions quickly reversible upon NE washout (16 +/- 9 min). This suggested that NE induction of long-term, but not short-term, plasticity in the dentate gyrus requires NMDA receptor activation. NE may be enhancing granule cell firing by some combination of blockade on the late Ca2+ activated K+ conductance and depolarization of granule cells, both actions that can bring granule cells into a voltage range where NMDA receptors are more easily activated. Furthermore, NE also elicited activity-independent long-lasting depolarization and RN increases, which required functional NMDA receptors to persist. PMID- 2572446 TI - Degeneration and graft-induced restoration of dopamine innervation in the weaver mouse neostriatum: a quantitative radioautographic study of [3H]dopamine uptake. AB - A recently introduced quantitative radioautographic technique was used to characterize the striatal dopaminergic deficit in weaver mutant mice and to evaluate the extent of DA reinnervation resulting from cell suspension grafts of fetal ventral mesencephalic tissue. Brain slices from normal mice and unilaterally grafted weaver mice were incubated in [3H]DA, in the presence of desipramine and pargyline 3-5 months after graft surgery. Semi-thin sections from the fixed and resinembedded slices were subsequently exposed on tritium sensitive film and afterwards dipped in nuclear emulsion for light microscope radioautography. Alternative slices were embedded in Epon for post-embedding tyrosine hydroxylase (TH) immunocytochemistry. The grain density of the film radioautographs matched well the distribution of TH positive fibers. Both methods revealed an almost complete absence of DA axons in the dorsomedial quadrant of the weaver neostriatum and an increasing density of DA innervation towards the ventrolateral areas. In the light microscope radioautographs, only the ventral striatum (i.e. nucleus accumbens and olfactory tubercle) and a narrow ventral and periventricular zone of the caudate-putamen were covered by silver grain clusters typical of DA varicosity labeling. Such labeled varicosities were nevertheless found in reduced numbers the lateral portion of both nucleus accumbens and the olfactory tubercle. The remaining neostriatum was overlaid by diffuse silver grains. suggesting a deficient DA uptake and storage mechanism in the residual DA fibers in this region. Immunocytochemistry using antibodies specific for DA or TH provided further evidence that the residual DA innervation in the weaver neostriatum was biochemically defective. Weaver mice with grafts of ventral mesencephalic tissue in the right neostriatum showed an amphetamine-induced rotational bias to the contralateral side, which was not seen in the sham operated animals. In contrast to the intrinsic weaver neostriatal DA innervation, DA fibers of graft origin exhibited the normal, clustered type of varicosity labeling. The computerized image analysis of silver grain density in film radioautographs was calibrated by counting these labeled varicosities in selected areas of light microscope radioautographs from the same sections. Results showed a mean DA reinnervation of neostriatal tissue surrounding the graft of about 20%, in some cases up to 80%, of the density seen in wild type mice, with a gradual decrease with distance up to 1-1.4 mm from the graft. The ventral parts of the neostriatum, which contained higher numbers of residual intrinsic DA fibers, were much more sparsely reinnervated than the dorsal and dorsomedial areas.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2572447 TI - Region specific regulation of glutamic acid decarboxylase mRNA expression by dopamine neurons in rat brain. AB - In situ hybridization histochemistry and RNA blots were used to study the expression of glutamic acid decarboxylase (GAD) mRNA in rats with or without a unilateral lesion of midbrain dopamine neurons. Two populations of GAD mRNA positive neurons were found in the intact caudate-putamen, substantia nigra and fronto-parietal cortex. In caudate-putamen, only one out of ten of the GAD mRNA positive neurons expressed high levels, while in substantia nigra every second of the positive neurons expressed high levels of GAD mRNA. Relatively few, but intensively labelled neurons were found in the intact fronto-parietal cerebral cortex. In addition, one out of six of the GAD mRNA positive neurons in the fronto-parietal cortex showed a low labeling. On the ipsilateral side, the forebrain dopamine deafferentation induced an increase in the number of neurons expressing high levels of GAD mRNA in caudate-putamen, and a decrease in fronto parietal cortex. A smaller decrease was also seen in substantia nigra. However, the total number of GAD mRNA positive neurons were not significantly changed in any of these brain regions. The changes in the levels of GAD mRNA after the dopamine lesion were confirmed by RNA blot analysis. Hence, midbrain dopamine neurons appear to control neuronal expression of GAD mRNA by a tonic down regulation in a fraction of GAD mRNA positive neurons in caudate-putamen, and a tonic up-regulation in a fraction of GAD mRNA positive neurons in fronto-parietal cortex and substantia nigra. PMID- 2572448 TI - Function of non-NMDA receptors and NMDA receptors in synaptic responses to natural somatosensory stimulation in the ventrobasal thalamus. AB - Sensory synaptic responses of rat ventrobasal thalamus neurones were challenged with iontophoretic applications of the excitatory amino acid antagonists CNQX and CPP. CNQX, applied with currents which were selective for non-NMDA receptors, antagonised responses of VB neurones to both 10 ms and 2000 ms air jet stimulation of the peripheral receptive field. In contrast, CPP only antagonised the latter type of response. These results suggest a differential involvement of excitatory amino acid receptors in sensory synaptic transmission to the ventrobasal thalamus, with an initial synaptic component being mediated by non NMDA receptors (including kainate receptors), and a further NMDA receptor mediated component being manifested upon maintained sensory stimulation. The expression of this latter component appears to be largely dependent upon the integrity of the non-NMDA receptor-mediated component. PMID- 2572449 TI - Astroglial cell alteration caused by neurotoxins: immunohistochemical observations with antibodies to glial fibrillary acidic protein, laminin, and tyrosine hydroxylase. AB - Kainic acid or 6-hydroxydopamine (6-OHDA) was injected into rat striatum, and their effects on astrocytes, laminin, and catecholamine fibers were examined temporally by immunohistochemical methods in an attempt to understand the roles of reactive astrocytes and laminin on the restoration of central nervous tissue. Kainic acid injection caused a severe neuronal degeneration in the striatum but catecholamine fibers were spared with only transient loss of tyrosine hydroxylase immunoreactivity. Reactive astrocytes appeared around the lesioned area soon after the kainic acid injection, then migrated into that area, and finally covered the lesioned striatum. Laminin immunoreactivity was found only in the lesioned area before the migration of reactive astrocytes and disappeared when the area was covered by astrocytes. 6-OHDA injection, on the other hand, resulted in a severe degeneration of catecholamine fibers, but striatal neurons were mostly spared. From 7 to 28 days after injection, regenerating fibers were found to enter the affected region. In this period reactive astrocytes were seen in the affected region but were only slightly more numerous than those found in control (saline injected) striatum. Laminin-immunoreactive blood vessels seemed to show a distribution similar to that in control striatum. These observations indicate that reactive astrocytes may play an important role in areas of neuronal cell loss and that laminin may aid their migration into such areas. Laminin and reactive astrocytes may not, however, be essential for the regeneration of dopamine fibers. PMID- 2572450 TI - Mutations in familial Creutzfeldt-Jakob disease and Gerstmann-Straussler Scheinker's syndrome. AB - A host protein encoded by the gene specifying the scrapie amyloid precursor affects pathogenesis of the transmissible spongiform encephalopathies: Creutzfeldt-Jakob disease (CJD), Gerstmann-Straussler-Scheinker's syndrome (GSS), and kuru in man, and scrapie in animals. We found a mutation in this gene of two patients with CJD from one family and a second mutation in the same gene in three patients with GSS from another family. The mutation in two related familial CJD patients changed glutamine in position 200 tolysine. This mutation was absent in other individuals including unrelated patients with familial CJD, sporadic CJD, and GSS. The other mutation in three GSS patients changed proline in position 102 to leucine, the same mutation described recently in some GSS families. We did not find it in six unaffected relatives of the GSS patients or in other individuals including sporadic and familial CJD patients. A rare insertion described earlier in one CJD family was also absent in all tested individuals. PMID- 2572452 TI - Possible correlation of TNF alpha-production with -RFLP in humans. PMID- 2572453 TI - Osmolality changes in nebulizer solutions. AB - Paradoxical effects (bronchoconstriction instead of bronchodilatation) have been reported after inhalation of beta2-mimetics in asthmatic children, and it has been suggested that this was due to osmolality and pH changes of the nebulizer solution. We tested commercially available nebulizer solutions and found osmolality changes after 5, 10 and 15 min of nebulization. Osmolality was measured in the nebulizer chamber and the airsteam of two types of jet nebulizers. When normal saline was nebulized chamber, from 282 +/- 7 mmol.kg-1 to 432 +/- 18 mmol.kg-1 resulted. Salbutamol ready made solution, and terbutaline respules were isotonic, whereas fenoterol, disodium cromogylcate (DSCG), beclomethasone dipropionate (BDP) and salbutamol (respirator solution 0.5%) were hypotonic (60-100 mmol.kg-1). When a mixture of sodium chloride (NaCl) and the drug solution (salbutamol, terbutaline, fenoterol) was nebulized for 10-15 min, the osmolality in the nebulizer cup increased to 420-500 mmol.kg-1. However, mixtures of the same beta 2-agonists with DSCG or with BDP remained hypo-osmolar. The same osmolality changes were present in the airstream. This study shows that after 10-15 min of nebulization osmotic changes occur in the nebulizer cup and airstream and that these changes differ according to the drug mixtures and the amount of the solution in the nebulizer chamber. PMID- 2572454 TI - [Effect of dipidolor on hemodynamics and respiration in rats after exposure to gamma-radiation and cytostatics]. AB - It was shown in experiments on anesthetized rats that intramuscular administration of dipidolor (5 mg/kg) failed to exert any effect on the functioning of the cardiovascular and respiratory systems of the animals which previously had been irradiated with the dose of 50 Gy or given intravenous injections of platidyam (5 mg/kg) or cyclophosphane (50 mg/kg). PMID- 2572451 TI - Melatonin biosynthesis in the mammalian pineal gland. AB - Rhythmic production of melatonin by the mammalian pineal occurs in response to noradrenergic stimulation which produces a cascade of biochemical events within the pinealocyte. In the rat, massive changes in NAT activity result from an increase in intracellular c-AMP levels produced by a synergistic interaction whereby an alpha 1 activation amplifies beta-adrenergic stimulation. The intracellular events mediating this effect are described. A major aspect of the temporal control of melatonin production is the programmed down-regulation of responses to noradrenergic stimulation once the initial surge of c-AMP is produced. Noradrenergic activation of the gland also influences other enzymic functions, including tryptophan hydroxylase and HIOMT activities, and produces a dramatic increase in intracellular c-GMP levels. Other neurotransmitters and neuropeptides, e.g. VIP, may also influence pineal function and comparisons are made between the rat, the subject of the bulk of experimental studies, and other species. PMID- 2572456 TI - Ca2+-dependent formation of an L-arginine-derived activator of soluble guanylyl cyclase in bovine lung. AB - In a fraction of cytosolic proteins from bovine lung, soluble guanylyl cyclase was concentration-dependently stimulated by L-arginine but not by D-arginine. Stimulation was up to 20-fold with an EC50 of about 3 x 10(-5) M. Activation of guanylyl cyclase by L-arginine was dependent on NADPH (EC50 about 5 x 10(-7) M) and Ca2+ (EC50 about 1.4 x 10(-6) M). The activation by L-arginine was inhibited by NG-monomethyl-L-arginine and hemoglobin. The effect of L-arginine was dependent on the protein concentration and was not observed in preparations of purified gyanylyl cyclase. These results suggest that bovine lung contains a Ca2+ regulated enzyme or enzyme system which converts L-arginine into an activator of soluble guanylyl cyclase. PMID- 2572455 TI - [Effect of drugs possessing antialcoholic activity on ethanol pharmacokinetics]. AB - Furazolidone, lithium carbonate, chlorprotixen and pyrroxan exert different effects on ethanol pharmacokinetics. Furazolidone decreases clearance of ethanol, increases its half-life with a simultaneous reduction of blood acetaldehyde concentration. Pyrroxan increases clearance and maximal concentration of ethanol. Chloprotixen effect is characterized by a concurrent increase of ethanol and acetaldehyde concentrations in the first hours after ethanol administration. Lithium carbonate produces no significant changes in ethanol pharmacokinetic parameters. PMID- 2572457 TI - Short and long spacer sequences and other structural features of zinc binding sites in zinc enzymes. AB - The crystal structures of eleven zinc enzymes have served to identify common features of their Zn binding sites. Two of them have non-catalytic Zn sites, both of which contain four cysteine ligands closely spaced in the linear sequence of the protein with no bound water. In contrast, all the catalytic Zn sites have three protein ligands and, in addition, one coordinated, 'activated' water. Histidine is the predominant ligand. The spacing between the first two ligands (1 3 amino acids), the short spacer, ensures a nucleus for Zn binding. The third ligand, separated by from approximately 20 to approximately 120 amino acids, the long spacer, not only completes the coordination but also aligns protein residues for interaction with the substrate. The short and long spacing observed for catalytic zinc sites may also pertain to Fe and Cu proteins. PMID- 2572458 TI - [The hygiene instruction and education of the workers of an animal-breeding farm complex in the activities of the feldsher at a feldsher-midwife center]. PMID- 2572459 TI - [Deontology in clinical research on children and adolescents]. PMID- 2572460 TI - Inflammatory bowel disease and male infertility: effects of sulfasalazine and 5 aminosalicylic acid on sperm-fertilizing capacity and reactive oxygen species generation. AB - To investigate the mechanism of the antifertility action of sulfasalazine, we prospectively measured sperm density, motility, hamster oocyte penetration capacity, and reactive oxygen species production in six men with inflammatory bowel disease being treated with sulfasalazine and 8 and 16 weeks after changing to 5-aminosalicylic acid (5-ASA). An improvement was observed after changing to 5 ASA in those with poor (n = 3), but not in those with normal (n = 3), sperm function on sulfasalazine. There was no correlation between reactive oxygen species production or acetylator phenotype with egg penetration capacity, sperm motility, or sperm density. We conclude that the mechanism of impairment and recovery of sperm function on and off sulfasalazine treatment is not related to an excess in reactive oxygen species. PMID- 2572461 TI - [The role of the endothelium in the beta-adrenergic regulation of vascular tonus]. AB - The effects of beta-adrenoreceptors agonists and antagonists on vascular tone were studied. Activation of beta-adrenoreceptors led to vasomotor reaction irrespective of the endothelium, through a direct effect on the smooth muscle elements of the vascular wall. The dilating effect of propranolol is determined by its interaction with endothelial cells which leads to the PGI2 and, perhaps, EDRF release. PMID- 2572462 TI - [The action of thyroliberin on the myoelectric activity of the stomach in the skate and the cod]. AB - The influence of exogenous thyrotropin-releasing hormone on the myoelectrical activity of stomach was investigated in skate and cod. The i.v. or intracerebral application of the peptide (5-8-10 micrograms/kg) led to an increase in the amplitude of the stomach cardiac and pyloric potentials, and to a change in their rate. The effect lasted longer following intracerebral administration of the peptide and was abolished by vagotomy. Irrespective of the technique of the administration, the peptide effect on the stomach myoelectrical activity was abolished by atropine and not affected by phentolamine or propranolol. PMID- 2572463 TI - Breeding habitats of mosquitoes in Goa. AB - Studies carried out in Goa from 1986 to 1987 revealed immature stages of 43 species of mosquitoes. Breeding habitats were divided into three categories, viz., (i) Ground water habitats consisting of ground pools and tanks, rocky pools, paddy fields and stream beds, yielding 30 species; (ii) Plant habitats consisting of tree holes and fallen coconut shells and leaf sheaths yielding 20 species; and (iii) Domestic/peridomestic habitats consisting of cement tanks, glass, earthen, metallic containers and tyres yielding 11 species. Five species viz., Anopheles subpictus, Aedes albopictus, Armigeres subalbatus, Culex quinquefasciatus and Cx. pallidothorax were found in all three types of habitats. Cx. tritaeniorhynchus preferred to breed in ground pools and paddy fields but was found in domestic cement tanks and ground tanks in adverse season. Majority of the anophelines and culicines were found in ground water habitats. Five out of 11 species of Aedes were found in tree holes. Aedes aegypti and Toxorhynchites splendens were found only in discarded tyres. Three species of the genus Uranotaenia, viz., U. bicolor, U. stricklandi and U. campestris were recorded from stream beds and plant containers. PMID- 2572464 TI - Immunoregulation by neuroendocrine factors. AB - The influence of the pituitary gland and of steroid hormones on the lymphoid system was demonstrated experimentally over half a century ago. Observations indicating the possibility of behavioural modification of immunity were also made at about the same time. Although these initial observations were followed by numerous investigations, the lack of sufficient basic knowledge of the endocrine and immune systems and serious methodological difficulties led to contradictions from which no definite conclusions could be drawn. Thus, the idea of neurohormonal-immune interaction fell gradually into disrepute. The remarkable effect of corticosteroids and their analogues on the immune system was regarded as a pharmacological phenomenon, rather than a physiological mechanism. During the past decade, this area gradually again became one of the forefronts of biomedical investigation and recently a number of volumes have been published on the subject of neurohormonal-immune interactions. Because of space limitations, only a brief overview of the subject can be given below and the reader is referred to the cited literature for detailed information. PMID- 2572465 TI - Peer review through perinatal statistics. AB - The purpose here is to demonstrate that perinatal statistics serve as a powerful tool for peer review, thereby improving the standard of maternity care. These data were used as an instrument for peer review, first by comparing perinatal mortality between one country and another. More detailed statistics, rapidly fed back to the delivery units, acted as a strong inducement to practitioners at all levels who wanted to do at least as well as their neighbors. Examples are given from Sweden, Scotland and Germany. PMID- 2572466 TI - Countrywide perinatal surveillance in Czechoslovakia. AB - Countrywide perinatal surveillance in four 10-year periods of Maternal Child Health (MCH) care development in Czechoslovakia is described. During this time maternal mortality decreased from 137 to 17/100,000 live births, and perinatal mortality (PMR) from 51 to 12/1000. On the basis of study results, countrywide intervention strategies concerning organization as well as medical measures were gradually drawn up. The greatest decrease in PMR was found through basic perinatal care for all; greatest efficiency followed employment of a risk approach. PMID- 2572467 TI - How Chinese clinicians contribute to the improvement of maternity care. AB - Clinicians in China are deeply involved in efforts to improve maternity care throughout the country. A Women's Health section has been established in the Chinese Society of Obstetrics and Gynecology. Staff of tertiary hospitals participate in perinatal surveillance of cities, including regular maternal and perinatal mortality meetings and comparative inter-hospital evaluations of service and clinical statistics. Clinicians from larger urban centers visit rural hospitals to provide guidance in management of high risk patients. PMID- 2572468 TI - Medical audit by peer review and competitive self-appraisal as tools for promoting efficiency: a case study in Lahore, Pakistan. AB - A pilot maternal and child health and family planning service project in urban and rural areas of Lahore, Pakistan, was evaluated after 4 years (1984-1988). In this time the birth rate declined from 41.0 to 19.2, infant mortality from 119.0 to 70.2, and maternal mortality from 560 to 220 per 100,000 live births. Success was attributed to medical audit of services through peer review of performance. PMID- 2572469 TI - Peer review among district health officers in Maharashtra, India. AB - A management information system in Maharashtra State, India, was used to obtain primary health care performance data for review at monthly meetings of peers, including District Health Officers, Auxiliary Nurse Midwives and Multipurpose Workers. The meetings were conducted in a problem solving, educational atmosphere. The effect of this process was marked improvement in health worker motivation and performance. PMID- 2572470 TI - Developing professional responsibility in maternity care in Hong Kong. AB - A simple questionnaire was used to document basic information surrounding all perinatal deaths in Hong Kong. Coordinators were appointed for every maternity unit. Replies were sent by mail to the Obstetrical and Gynaecological Society. Using volunteer time and minimum budget, the data were collated and analyzed. The main causes of death were congenital anomalies, intrauterine death of unknown cause and prematurity. There was significant difference in performance when the University units were compared with the private and Government units. Changes in resource allocation and modification of clinical practice may be recommended as a result of such audit. PMID- 2572471 TI - The role of confidential enquiries in the reduction of maternal mortality and alternatives to this approach. AB - The aim of confidential enquiries into maternal deaths is to identify weaknesses in the maternal health care system with a view to remedying them. The method of confidential enquiry is explained using the British system as an example. The reasons why this apparently useful practice is not more widely adopted can in some countries include fears of litigation or lack of trust in confidentiality. Alternative approaches to maternal death audit are discussed. PMID- 2572473 TI - The perinatal survey of the south-west region in the United Kingdom. AB - The South Western Region of the United Kingdom (UK) has a population of over three million and approximately 38,000 births each year in 48 maternity units. During 1980-1982, a detailed survey, including site visits, was undertaken of the perinatal facilities, staffing, workload and outcome. During that period the perinatal mortality for the region fell faster than in any other region; at 9.1 per 1000 births in 1982, it was also the lowest for any region in the UK. PMID- 2572472 TI - Professional responsibility in maternity care: role of medical audit. AB - In 1965, Baroda Medical College initiated a process of medical audit of maternal and perinatal deaths occurring at this institution, and consultation in peripheral medical facilities providing antenatal and obstetric care. By 1984 maternal and perinatal mortality had declined and clinical judgment in maternity care had improved. PMID- 2572474 TI - Homozygous sickle cell anemia at the University College Hospital, Ibadan, revisited. AB - The outcome of 78 pregnancies in 47 patients with homozygous sickle cell anemia managed at the University College Hospital, Ibadan between January 1, 1975 to December 31, 1984 is reviewed. There were two maternal deaths giving a maternal mortality rate of 25 per thousand. Abortion rate was 155 per thousand. The overall perinatal mortality rate was 188 per thousand. Forty-five percent of the babies were of low birthweight and 29% were preterm. Anemia was the commonest antenatal complication and the highest indication for hospitalization. The data were compared with previous ones from this hospital and with similar reports by other investigators outside Nigeria. A need for meticulous antenatal care and prenatal diagnosis to eliminate the homozygous sickle cell anemia is stressed. PMID- 2572475 TI - Oral versus local treatment of vaginal candidosis. AB - A comparative study was conducted to compare the results of the use of oral and local vaginal therapy in the treatment of vaginal candidosis. Forty-five patients with clinical and mycological evidence of vaginal candidosis were recruited and were randomly allocated to one of the treatment groups. Twenty-four patients received ketoconazole orally (400 mg/day for 5 days) and 21 patients received nystatin vaginal pessaries (100,000 units twice/day for 7 days). Seven out of 20 rectal swabs (35%) were positive for Candida albicans. Both drugs were significantly effective in relieving patients symptoms and physical signs. The mycological cure rates were 87.5% and 81%, and the relapsing rates were 0% and 5.9% for the ketoconazole and nystatin groups, respectively. Ketoconazole oral therapy had generally slightly higher results than local nystatin therapy in the treatment of vaginal candidosis, yet the difference was statistically insignificant. PMID- 2572476 TI - Menarcheal age in Ghanaian school girls. AB - In a cross-sectional retrospective study of 2087 Ghanaian school girls in various educational institutions in the Kumasi district, Ashanti region, Ghana, the mean menarcheal age was found to be 13.98 +/- 1.42 years. Differences in the menarcheal age of the girls was found to be significantly correlated to social class, parents ethnic origin, educational institution and home living area (P = 0.0001). The duration of the menarche and the interval between the menarche and the second period was found to be influenced by the age at menarche (P less than 0.01). Decline in menarcheal age in concurrence with world trends was observed. Further studies are necessary to identify the inherent and specific factors in the Ghanaian population which relate to and influence the age of menarche. PMID- 2572478 TI - Conjoined twins and abdominal pregnancy. AB - Abdominal pregnancy is rare; conjoined twins are also rare. The diagnosis of abdominal pregnancy is easily missed and often not made until near full-term. Similarly conjoined twins are not diagnosed until late in gestation or during parturition. In this case, the diagnosis of conjoined twins and abdominal pregnancy was only made during laparotomy. PMID- 2572477 TI - Fentanyl-midazolam-flumazenil anesthesia for induced abortion. AB - A new anesthetic method (fentanyl-midazolam-flumazenil) was compared with a previously administered anesthetic regimen (pethidine-diazepam-ketamine) in two groups of 25 women, each of whom underwent termination of pregnancy. No significant difference was found between the two groups in quality of anesthesia. Recovery was assessed by means of the Aldrete score and a visual analog scale. The recovery time was significantly shorter in patients who received the fentanyl midazolam-flumazenil. PMID- 2572479 TI - Phosphatidylglycerol, lecithin/sphingomyelin ratio and respiratory distress syndrome in diabetic and non-diabetic pregnancies. AB - Samples of amniotic fluid from 514 non-diabetic and 69 diabetic patients were analyzed for phospholipid content. Results were correlated with incidence of respiratory distress syndrome (RDS) in the neonate. The incidence of RDS was 4.5% among diabetics and 5.3% among non-diabetics. In the presence of phosphatidylglycerol (PG), no infant developed RDS while in the absence of PG the incidence of RDS was 16.7% and 14.4%, respectively. In the presence of a mature lecithin/sphingomyelin (L/S) ratio the respective incidence of RDS was 1.6 and 1.8, while with an immature L/S ratio the incidence of RDS was 28.6% and 29%. The incidence of RDS after 37 weeks gestation was 0% among diabetics and 0.6% among non-diabetics. We conclude that amniotic fluid phospholipids are equally predictive of risk for RDS in diabetics as among non-diabetic patients. We suggest that in patients with accurate gestational dating, amniotic fluid analysis for phospholipids might not be necessary to establish fetal lung maturity. PMID- 2572480 TI - Microprocessor based waveform analysis of the fetal electrocardiogram during labor. AB - An increase in T wave amplitude of the fetal ECG (FECG) has experimentally been correlated to elevated catecholamine levels and myocardial glycogenolysis. The FECG changes have also been described during human delivery. The present aim was to investigate whether these ECG changes could be reproduced in an easily handled real time microprocessor system, and to correlate them to biochemical and clinical data. During 40 deliveries the FECG signal was transferred to a microcomputer system for real time averaging of the FECG. There was a high capacity of the system to reproduce the ST waveform changes though the averaging procedure reduced the QRS magnitude by 10%. With a normal umbilical artery pH (greater than or equal to 7.25) the highest T/QRS ratios recorded during each delivery was 0.26 +/- 0.19 (mean +/- S.D.). With lowered pH (less than 7.25) the T/QRS increased to 0.33 +/- 0.10 (P less than 0.02). A similar difference between the two groups was seen when the T/QRS ratios from the last hour before birth were compared; 0.13 +/- 0.08 and 0.18 +/- 0.05, respectively (P less than 0.01). Changes in the ST waveform with T/QRS greater than or equal to 0.30, ST segment alterations, or negative T waves appeared during 40% of the deliveries, however, 30% were short standing changes (less than 30 min). Intermediate CTG changes during at least 30 min occurred in 41% and the pattern was classified as abnormal in 18%. Using the scalp electrode as signal source, the ECG analysis could add further information to the routine CTG recording on the fetal condition during delivery. PMID- 2572481 TI - A sixteen year survey of maternal mortality associated with eclampsia in Ilorin, Nigeria. AB - An analysis was made of 94 maternal deaths following eclampsia, occurring over a 16-year period. Maternal deaths were high in the very young, primigravida and in elderly women of higher parity. The overall maternal mortality over the years appears the same; while mortality associated with eclampsia seems dependent on time. Factors attributed to high maternal death rate from eclampsia are examined and recommendations made as to their prevention. PMID- 2572483 TI - Burn injuries during the puerperium in Zaria, Nigeria. AB - Of the 35 women who sustained burn injuries during the puerperium in Zaria, 31 had practised ritual hot baths or "Wankan-jego" which caused severe scalds. Five of the women also developed peripartum cardiac failure. Two died. Three of the newborns also died partly as a result of the mothers' illness. Formal education and general economic development would help in eliminating this preventable source of maternal and infant morbidity and mortality. PMID- 2572482 TI - A study of antenatal care at village level in rural Tanzania. AB - Antenatal care is an acknowledged measure for the reduction of maternal and perinatal mortality. In the rural village of Ilula, Tanzania, the possible impact of antenatal care on mortality was studied longitudinally on the basis of the 707 women delivered in the study period. Ninety-five percent of the antenatal records were available. Anemia, malaria and anticipated obstetric problems were the most frequent reasons for interventions. Among the women from the area who were delivered in hospital, 90% had been referred there. No relationship was found between the number of antenatal visits and the pregnancy outcome, but perinatal mortality was correlated to a low birth weight. Even with a mean attendance rate of six visits and full coverage by antenatal care maternal and perinatal mortality remains high. PMID- 2572484 TI - Should the Cytobrush be used in routine screening for cervical pathology? AB - In 632 women visiting a family planning clinic, paired cervical smears were taken using a wooden spatula and a Cytobrush. Of the spatula smears 7.8% were unreliable with too little or scanty material to allow a cytologic diagnosis (class 0); 17.7% of spatula smears were inadequate, lacking endocervical cells. Combined spatula/Cytobrush cytology reduced these figures significantly; 4.3% unreliable smears and 3.2% inadequate specimens. Seventy-three (11.6%) of the smears revealed some abnormality, in most cases Papanicolaou class IIIA, suggestive of mild dysplasia. In 10 cases, dysplastic cells were found only in the Cytobrush cytology. It is concluded that the Cytobrush adds to the improvement of smear quality and the accuracy of routine cervical cytology obtained by relatively inexperienced smear takers. PMID- 2572485 TI - Hormonal and basement membrane markers for immunoidentification of cultured human trophoblast cells. AB - Improvement of human chorionic villi cells in vitro culture has been obtained by supplementing the medium with 30% human fetal cord serum, instead of fetal calf serum. Expression of human chorionic gonadotropin, estrogen and progesterone receptors, laminin, laminin receptors, and type IV collagen has been studied on first subculture passages (4-7 weeks) by immunoperoxidase method. A minority of the cultured cells were positive for estrogen receptors, the majority were positive for progesterone receptors, while all cells were negative for human chorionic gonadotropin. Cultured cells showed variable positive immunostaining for basement membrane markers like laminin, and type IV collagen, and for laminin receptors. Detection of both progesterone receptors and laminin, or type IV collagen, excluded fibroblast contamination and could then be useful for quick identification of cultured trophoblast cells. PMID- 2572486 TI - Endometrial flora of infertile women in Zaria, northern Nigeria. AB - Microbiological study of the endometrium of 114 infertile women cultured 48 organisms (42.2%) from 42 patients (36.8%). Mycobacterium tuberculosis (16.7%) and Group B Streptococcus (8.8%) were the commonest organisms isolated. These organisms were also most commonly isolated from patients between the ages of 20 and 30 years. There is, however, an overall increase in the frequency of bacterial isolation with age-cohorts. Of the colonised patients, 36.8% were of parity less than two and most (26.3%) were nulliparous. It is possible that multiple marriages and polygamy played a significant role in the bacterial colonisation of the endometrium in the Hausa-Fulani population of Zaria, Nigeria. PMID- 2572487 TI - Uterine unification procedures: post-operative obstetrical outcome. AB - Seventeen patients with different uterine anomalies underwent unification procedures for various indications. Preoperative obstetrical performance segregated them into three main groups: seven patients had only first trimester abortion (group A), seven patients had second trimester abortion with or without first trimester abortion (group B) and three patients were infertile (group C). Postoperatively, all seven patients in group A became pregnant with 100% viable pregnancy rate. Six patients in group B became pregnant (85.7%) and five (57%) had living children. All three patients with primary infertility (group C) became pregnant and two had living babies. The overall pregnancy rate was 94.1% and the viable pregnancy rate was 76.5%. Uterine unification procedures are recommended for patients with repeated first trimester abortions as well as for selected patients with infertility. PMID- 2572489 TI - Fetal resuscitation in a patient with varicella pneumonia and preterm labor. AB - A primigravida presented at 33 weeks gestation with varicella pneumonia. Cardiotocography demonstrated repeated late decelerations of fetal heart rate. Cesarean section was indicated for contracted maternal pelvis and breech presentation, but was decided against because of the risks of anesthesia. Maternal and fetal improvement followed intrauterine resuscitation and suppression of preterm labor. Cesarean section was performed 9 days later after spontaneous rupture of membranes with excellent outcome. The same approach is suggested for similar cases. PMID- 2572488 TI - A comparative study of Norinyl 1/35 versus Lo-Ovral in Ile-Ife Nigeria. AB - A comparative analysis of those women on Norinyl 1/35 versus those on Lo-Ovral showed those on Norinyl 1/35 were significantly (P less than or equal to 0.05) older than women in the Lo-Ovral group. There were no significant differences (P greater than or equal to 0.05) between the groups for other characteristics at admission. There were no significant differences (P greater than or equal to 0.05) between the groups at follow-up in the number of women reporting serious complications, menstrual complaints or other pill-related problems. The continuation rates at 12 months were 79.4 for the Norinyl 1/35 group and 76.7 for the Lo-Ovral groups. There were no pregnancies reported during the study period. PMID- 2572491 TI - Antenatal diagnosis of acardiac, acephalic twin. AB - A case of acardiac, acephalic twin diagnosed by antenatal ultrasound is reported. The obstetrical significance of this condition is discussed. PMID- 2572490 TI - Control of prostaglandin induced uterine hyperactivity with intravenous ritodrine. AB - The prostaglandins, F2 alpha and E2, are in extensive local use for the induction of labor. The main concern with oral and vaginal administration is the difficulty in controlling rapidly progressing labor and uterine hyperactivity. Herein we present a case where intravenous ritodrine was given as soon as hyperactivity and fetal heart rate decelerations were detected. With this treatment, the hyperactivity was controlled, fetal heart rate returned to normal and labor progressed normally to the birth of a healthy infant. PMID- 2572493 TI - Natural transmission of group B Streptococcus during delivery. AB - This is a study of group B Streptococcus during labor of 121 patients in whom group B Streptococcus was isolated in the vagina and/or rectum before delivery. The intrapartum vaginal culture was positive in 55.2% of the antepartum carriers (32/58). When the vaginal culture during delivery was positive, the group B Streptococcus was isolated in the amniotic fluid 2 h after the rupture of membranes in 81% of the cases. The newborns of antepartum carriers, when the labor developed naturally, were colonized by group B Streptococcus in 69.2% of cases when the intrapartum vaginal and/or amniotic fluid cultures were positive (9/13), while only 5.6% of the newborns of antepartum carriers but with negative cultures during delivery were colonized by group B Streptococcus (1/18). The most frequent positive neonatal culture was in the umbilicus (83.3%) followed by the external ear (62.5%). PMID- 2572492 TI - Cesarean and postpartum hysterectomy in Enugu, 1973-1986. AB - Obstetric hysterectomies were performed on 163 patients over a period of 13 years out of 56,823 deliveries. Seventy-one percent of the patients received no antenatal care in the index pregnancies. The majority were of high parity. Ruptured uterus was the commonest indication. Inadequate transfusion and late presentation affected the prognosis adversely. Ignorance, availability of suitable maternity care units and uncontrolled reproductive patterns were the main predisposing factors. PMID- 2572494 TI - Etizolam in the treatment of generalized anxiety disorder: a controlled clinical trial. AB - A total of 45 patients with generalized anxiety disorder were treated twice daily for 2 weeks, on a double-blind basis, with 0.5 mg etizolam, 0.5 mg alprazolam or 3 mg bromazepam, and symptoms were assessed using Hamilton's rating scale for anxiety and Hamilton's rating scale for depression. Patients then received the same drug for a further 2 weeks, the drugs being given three times daily if a poor response was observed during the first 2 weeks. All drugs displayed equivalent anxiolytic activity after 2 weeks, but etizolam displayed a progressive increase in anxiolytic activity over 4 weeks of treatment. Etizolam also possessed a more marked antidepressant effect than did alprazolam or bromazepam. There were no differences in the tolerability of the three drugs. PMID- 2572495 TI - Postoperative recurrence of Crohn's disease in relation to radicality of operation and sulfasalazine prophylaxis: a multicenter trial. AB - Recurrence rate is high after operation for Crohn's disease. A multicenter trial was performed to study the effect of radical or nonradical operation and of sulfasalazine prophylaxis versus placebo on postoperative recurrence rate in 232 patients with Crohn's disease. Sixteen medical and surgical centers participated in the study, 7 operating radically and 9 nonradically. The follow-up period lasted 3 years, the allocation to drug treatment was randomized and double blind. Recurrence was significantly less frequent and occurred later in patients who were operated nonradically. Patients on sulfasalazine prophylaxis had a better prognosis than on placebo. This effect was statistically significant in the first 2 years of treatment. Both strategies were additive: nonradical operation and sulfasalazine had the best prognosis, radical operation and placebo was worst. It is concluded that postoperative recurrence is best prevented by resecting nonradically and prescribing 3 g of sulfasalazine daily at least over 2 years. PMID- 2572496 TI - Treatment of patients with chronic type B hepatitis and concurrent human immunodeficiency virus infection with a combination of interferon alpha and azidothymidine: a pilot study. AB - Six patients with chronic type B hepatitis and concurrent infection with the immunodeficiency virus were treated with 600 mg azidothymidine (AZT)/day and 3 X 10(6) units of interferon-alpha (IFN-alpha) every other day for a total of 4 months. None of the patients treated lost the hepatitis B virus (HBV). HBV-DNA concentrations were not significantly influenced by this treatment. Human immunodeficiency virus (HIV) infection was also not affected except for a transient rise in CD 4-positive cells in 2 individuals, who had initially low CD 4-positive cells. Treatment did not influence the presence of HIV-Ag in the serum. In conclusion, a combination therapy of IFN and AZT does not seem to be beneficial at the doses given and the time involved. PMID- 2572497 TI - Effect of neural blockade on somatostatin-induced inhibition of exocrine pancreatic secretion. AB - In vivo somatostatin inhibits exocrine pancreatic secretion. In in vitro experiments, such as the isolated perfused pancreas, somatostatin fails to inhibit exocrine pancreatic secretion. This suggests an indirect action of somatostatin, such as modulation of neural regulation. In the anesthetized rat we tested the inhibitory capacity of somatostatin in the presence of neural blockade in vivo. Neither the drugs given intravenously--phentolamine, propranolol, atropine and naloxone--nor vagotomy were able to prevent somatostatin-induced inhibition of exocrine pancreatic secretion. We conclude that somatostatin induced inhibition of exocrine pancreatic secretion is not dependent on intact extrinsic innervation. PMID- 2572498 TI - TNF-alpha gene polymorphisms in type 1 (insulin-dependent) diabetes mellitus. AB - Type 1 (insulin-dependent) diabetes mellitus, like some other autoimmune diseases, is linked to certain alleles coded by genes in the HLA-D region. Sequence analysis of DQ beta chains indicates that aspartic acid at codon 57 confers resistance to the development of Type 1 diabetes. However, a full explanation for the HLA-association of Type 1 diabetes, particularly the increased susceptibility of DR3/4 heterozygotes is still awaited. The localisation of tumour necrosis factor genes on the short arm of chromosome 6 between HLA-B and the complement genes (Class III) prompted us to investigate a possible polymorphism of TNF-alpha at the genomic level in relation to Type 1 diabetes susceptibility. A dialleleic TNF-alpha restriction fragment length polymorphism was found with Ncol and its segregation with HLA-haplotypes analysed in diabetic families. We describe here a strong linkage of TNF-alpha alleles with certain DR haplotypes. For example, the common extended haplotype HLA A1-B8-DR3 was almost exclusively associated with the 5.5 kb TNF-alpha allele whereas Bw62 DR4 with the 10.5 kb allele. Thus both alleles segregate to diabetic patients. DR matched haplotypes of affected family members differed significantly from those of the non-affected at the TNF alpha locus. All affected sibling pairs in 11 multiplex affected families were identical for TNF-alpha alleles, even if they were only haploidentical for HLA-B-DR haplotypes. In addition, heterozygosity for the TNF-alpha alleles was significantly more frequent in the patients. This tight linkage of TNF-alpha alleles with some extended haplotypes could help to explain the HLA-association of Type 1 diabetes as well as some other autoimmune diseases. PMID- 2572499 TI - Antigen presentation. AB - This paper reviews some of the cellular events involved in the immune recognition of foreign proteins. The recognition of an antigen by T lymphocytes is essential for its effective elimination by the host. T lymphocytes of the CD4 or CD8 subset recognize antigen but only after the antigen is handled by antigen-handling cells (antigen-presenting cells). Antigen molecules are recognized after an internal processing event by antigen-presenting cells that results in the generation of immunogenic peptides. Such peptides associate with histocompatibility molecules to form bimolecular complexes on the cell surface. The T cell receptors for antigen recognize the bimolecular complex and initiate the events that result in an inflammatory response. Antigen-presenting cells also produce molecules - termed costimulators - that stimulate the growth and differentiation of T lymphocytes. PMID- 2572500 TI - A role for Na+-dependent Ca2+ extrusion in protection against neuronal excitotoxicity. AB - The hypothesis that Na+-dependent calcium extrusion is important in protecting against neuronal excitotoxicity was tested. In cocultures of embryonic rat hippocampal neurons and mouse neuroblastoma hybrid (NCB-20) cells, calcium ionophore A23187 (1 microM) or high levels of extracellular K+ killed hippocampal neurons selectively, leaving NCB-20 cells unscathed. Hippocampal neurons showed large, sustained rises in intracellular calcium in response to A23187 or K+, whereas NCB-20 cells showed only transient calcium responses. The ability of NCB 20 cells to reduce the calcium load and to survive exposure to A23187 or K+ were dependent on extracellular Na+, suggesting that an active Na+/Ca2+ exchange mechanism was important in protecting against cell death. Finally, removal of extracellular Na+ reduced the threshold for glutamate neurotoxicity in hippocampal neurons, demonstrating the importance of Na+/Ca2+ exchange in protecting against excitotoxicity. Taken together, these findings suggest that differences in cell calcium-regulating systems may determine whether a neuron lives or degenerates in the face of an excitatory challenge. PMID- 2572501 TI - Glutathione metabolism at the blood-cerebrospinal fluid barrier. AB - Glutathione metabolism and transport in the choroid plexus were probed by determining the effects of administration to rats of several compounds (buthionine sulfoximine, L-2-oxothiazolidine-4-carboxylate, L-(alpha 5,5S)-alpha amino-3-chloro-4,5-dihydro-5-isoxazole acetic acid, gamma-glutamyl alanine, and glutathione monoethyl ester) on the levels of glutathione and cysteine in the cerebrospinal fluid. The findings indicate that glutathione is actively metabolized in the choroid plexus by pathways similar to those in kidney and other tissues. The level of glutathione in the cerebrospinal fluid can be decreased or increased by giving compounds that do not, under similar conditions, appreciably alter total brain levels of glutathione. Glutathione monoethyl ester is effectively transported into the cerebrospinal fluid. PMID- 2572502 TI - This is FASEB. The American Society for Cell Biology. PMID- 2572503 TI - [Role of the adrenergic system in atherogenesis and effects of beta-blockers]. PMID- 2572504 TI - [Sedation in digestive endoscopy]. PMID- 2572505 TI - Gastric acid secretion stimulated by modified sham-feeding, and the effects of histamine H2-antagonist and anti-muscarinic agent in patients with duodenal ulcer. AB - Patients with duodenal ulcer (DU) were classified into responders to H2 antagonist and non-responders in whom DU did not heal within 3 months with the antagonist. In these patients and healthy controls, a modified sham-feeding (MSF) test was performed to elucidate the pathogenesis of resistance to H2-antagonist. By MSF stimulation, gastric acid secretion significantly increased in all subjects. The mean acid output by MSF amounted to about 52 of the tetra-gastrin maximum in controls (12 cases), 46% in responders (12 cases) and 72% in non responders (14 cases). The mean acid output in non-responders was significantly higher than in controls under baseline conditions, MSF and gastrin stimulations, but was higher than in responders during MSF stimulation. The effects of H2 antagonist (cimetidine 1mg/kg/h), anti-muscarinic agent (pirenzepine 0.3mg/kg/h), or both on the acid secretion were examined on non-responders (6 cases), responders (6 cases) and healthy controls (6 cases). The acid secretion stimulated by MSF was significantly inhibited by pirenzepine in responders and controls, but not in non-responders. With cimetidine, the acid output was significantly inhibited in all groups, but was still higher in non-responders than in controls and responders, indicating that the reduction of acid output by a H2-antagonist is significantly less in non-responders than in other groups. The combined use of pirenzepine and cimetidine almost completely inhibited the acid output in all groups. These data suggest that the vagal activity in non responders to H2-antagonist is higher than in responders and healthy subjects, and H2-antagonist combined with anti-muscarinic agent is more effective in reducing the gastric acid secretion in non-responders. PMID- 2572506 TI - Somatostatin inhibition of thyrotropin-releasing hormone- and growth hormone releasing factor-induced growth hormone secretion in young and adult anesthetized chickens. AB - The present communication examines the influence of somatostatin (SRIF14) on basal and thyrotropin-releasing hormone (TRH)- or growth hormone-releasing factor (GRF)-induced GH secretion in young (6 week old) and adult male chickens. Studies were performed in sodium pentobarbitone-anesthetized chickens where basal plasma concentrations of GH are low and both TRH and GRF consistently stimulate GH release. In both young and adult chickens, basal GH secretion was reduced by SRIF14 infusion (3 micrograms/kg/min). Similarly, in adult and young birds, the GH secretory response to a challenge with a GRF bolus (10 micrograms/kg) was inhibited by the concomitant intravenous infusion of SRIF14 (0.3 or 3.0 micrograms/kg/min). In adult chickens, the GH response to TRH (10 micrograms/kg) was suppressed (by 93%) by SRIF14 infusion (3 micrograms/kg/min) and tended to be inhibited (by 29%) by a lower dose of SRIF14 (0.3 micrograms/kg/min). In contrast, in young chicks, GH release following TRH challenge (either 1 or 10 micrograms/kg) was only partially inhibited by SRIF14 infusion (3 micrograms/kg/min). The response to a TRH challenge (10 micrograms/kg) was unaffected by the low dosage of SRIF14 infusion (0.3 micrograms/kg/min). It is concluded that SRIF14 inhibits both GRF- and TRH-stimulated GH release in young and adult chickens. PMID- 2572507 TI - Sequence analysis of N-ethyl-N-nitrosourea-induced vermilion mutations in Drosophila melanogaster. AB - The mutational specificity of N-ethyl-N-nitrosourea (ENU) was determined in Drosophila melanogaster using the vermilion locus as a target gene. 25 mutants (16 F1 and 9 F2 mutants) were cloned and sequenced. Only base-pair changes were observed; three of the mutants represented double base substitutions. Transition mutations were the most prominent sequence change: 61% were GC----AT and 18% AT-- -GC substitutions. Both sequence changes can be explained by the miscoding properties of the modified guanine and thymine bases. A strong bias of neighboring bases on the occurrence of the GC----AT transitions or a strand preference of both types of transition mutations was not observed. The spectrum of ENU mutations in D. melanogaster includes a significant fraction (21%) of transversion mutations. Our data indicate that like in other prokaryotic and eukaryotic systems also in D. melanogaster the O6-ethylguanine adduct is the most prominent premutational lesion after ENU treatment. The strong contribution of the O6-ethylguanine adduct to the mutagenicity of ENU possibly explains the absence of distinct difference between the type of mutations observed in the F1 and F2 mutants. Although the latter arise later during development, the spectrum of mosaic mutations is also dominated by GC----AT transition mutations. PMID- 2572508 TI - A molecular genetic linkage map of mouse chromosome 9 with regional localizations for the Gsta, T3g, Ets-1 and Ldlr loci. AB - A 64-centiMorgan linkage map of mouse chromosome 9 was developed using cloned DNA markers and an interspecific backcross between Mus spretus and the C57BL/6J inbred strain. This map was compared to conventional genetic maps using six markers previously localized in laboratory mouse strains. These markers included thymus cell antigen-1, cytochrome P450-3, dilute, transferrin, cholecystokinin, and the G-protein alpha inhibitory subunit. No evidence was seen for segregation distortion, chromosome rearrangements, or altered genetic distances in the results from interspecific backcross mapping. Regional map locations were determined for four genes that were previously assigned to chromosome 9 using somatic cell hybrids. These genes were glutathione S-transferase Ya subunit (Gsta), the T3 gamma subunit, the low density lipoprotein receptor, and the Ets-1 oncogene. The map locations for these genes establish new regions of synteny between mouse chromosome 9 and human chromosomes 6, 11, and 19. In addition, the close linkage detected between the dilute and Gsta loci suggests that the Gsta locus may be part of the dilute/short ear complex, one of the most extensively studied genetic regions of the mouse. PMID- 2572509 TI - Voluntary and involuntary schizophrenic patient admissions on the same general hospital psychiatric unit. AB - The authors compare a group of voluntary schizophrenics to another group of involuntary schizophrenic patients with regard to their various clinical and management aspects. Both groups had been treated on the same unit, by the same staff, and within the same approximate time period. The involuntary group needed a slightly longer stay, a significantly higher dosage of neuroleptics, and more staff supervision and intervention. The involuntary schizophrenic patients were more likely to be violent, with agitation and destructive behavior, and to require restraints or seclusion. PMID- 2572510 TI - [Study of polymorphism in human rRNA gene clusters]. AB - Human ribosomal DNA (rDNA) probe specific for the 3' end of the 28S rRNA gene was used for detecting standard restriction fragments' length polymorphism (RFLPs) in the non-transcribed spacer. The conditions for hybridization of rDNA probe which eliminate cross hybridization of parts of 28S rRNA gene were developed. A test for detecting incompletely restricted DNA was also developed which may be used in experiments for detecting new RFLPs. It was found that a set of standard RFLPs was identical in various human tissues for one individual. Frequency of standard RFLPs in the non-transcribed spacer of human rRNA gene clusters was calculated. PMID- 2572511 TI - cDNA sequence and chromosomal localization of the mouse parvalbumin gene, Pva. AB - In the homozygous condition, the mutation adr (arrested development of righting response) of the mouse causes a myotonia and a drastic reduction of the Ca2+ binding protein parvalbumin (PV) in fast muscles. Using a rat PV probe, a mouse cDNA clone was isolated from a lambda gt11 wild-type fast-muscle library and its nucleotide sequence was determined. The protein coding and the 3' nontranslated regions of the mouse gene show extensive homology with the rat PV gene. The result of Southern blot hybridization is consistent with a single copy gene for parvalbumin. Restriction fragment length polymorphisms (RFLPs) between Mus musculus domesticus (e.g. C57BL/6) and Mus spretus (SPE) were detected with the enzymes Eco RI, Pst I, and Sst I. The restriction fragment patterns of DNA samples from 65 individual offspring of (C57BL/6 x SPE)F1 x C57BL/6 backcrosses were tested with the PV probe and matched, for linkage detection, to pre-existing patterns established with various RFLP probes on the same samples. A co distribution of PV-RFLPs with Pvt-1 and Mlvi-2, which had been localized on chromosome 15, was detected. Thus, the structural gene for PV, designated Pva, maps to chromosome 15 of the mouse whereas the adr mutation shows no linkage with markers on this chromosome. Gene locus homology between chromosome 15 of the mouse and chromosome 22 of man (which carries the human PV gene) is discussed. PMID- 2572512 TI - The structure of the chicken glutamine synthetase-encoding gene. AB - Complementary DNA (cDNA) and genomic clones encoding chicken glutamine synthetase (Glns) have been isolated. The nucleotide (nt) sequence of the 2728-bp cDNA specifies a 91-nt 5' untranslated sequence, a 1119-nt open reading frame, and a 1518-nt 3' untranslated sequence that contains several A + T-rich regions but lacks a canonical endonucleolytic-cleavage/polyadenylation signal. Based on sequence analysis of the cloned gene, the Glns transcription unit spans 7.0 kb and contains seven exons. PMID- 2572513 TI - The activity of the Pseudomonas aeruginosa pilin promoter is enhanced by an upstream regulatory site. AB - Recently, a comparison of the nucleotide sequences upstream of the structural pilin genes (pil) from five different isolates of Pseudomonas aeruginosa identified a sequence homologous to a NifA-binding site, a positive regulatory sequence in Klebsiella pneumoniae which controls the expression of the genes which fix nitrogen [Pasloske et al., J. Bacteriol. 170 (1988) 3738-3741]. This sequence was located between -98 and -114 bp relative to the transcription start point. To establish whether this putative recognition sequence exerted any regulatory effect on pil gene expression, we assayed for the expression of pilin and pili, and assessed the strength of the pilin promoter with and without this upstream sequence. A 48-bp deletion, which removed the putative recognition sequence, decreased the levels of pilin and pilus expression. Also, the equivalent deletion reduced pilin promoter activity by 90% when it was measured using a promoter probe vector. These results provide evidence that a nucleotide sequence upstream of the pilin promoter positively regulates the transcription of the P. aeruginosa pil gene. PMID- 2572514 TI - Antihypertensive drugs: adverse reactions and metabolic effects in elderly patients. AB - Symptomatic and metabolic side effects seen in the treatment of elderly hypertensive patients are an area of concern for physicians. Of the four drug classes used as Step 1 antihypertensive agents--diuretics, beta-blockers, ACE inhibitors, and calcium-entry blocking agents--only diuretics have been studied as monotherapy for the elderly in placebo-controlled, long-term clinical trials. These studies have shown diuretics to be well tolerated in this patient population, producing only mild, infrequent side effects. PMID- 2572515 TI - Pharmacologic treatment of the elderly hypertensive: comparative efficacy and side effects. AB - Because of the physiologic changes associated with aging, a nonpharmacologic approach is recommended as initial therapy for the elderly hypertensive. Approximately 80% of this patient population, however, requires pharmacologic therapy, as well. When initiating drug treatment, the physician should remember several guidelines and have some understanding of the mechanisms of action of the available agents. The four drug classes recommended by JNC IV for the treatment of high blood pressure are quite similar in effectiveness. Low-dose diuretics, however, particularly when combined with a potassium-sparing agent, produce significantly fewer adverse reactions than seen with the other classes of drugs. This advantage, coupled with their low cost, makes diuretics the reasonable first choice for the treatment of older hypertensives. PMID- 2572516 TI - Histidine and proline are important sites of free radical damage to proteins. AB - Our hypothesis that proline and histidine are major sites of damage during radical attack upon proteins, becoming respectively glutamate and aspartate, was investigated using proteins biosynthetically labelled with radioactive proline or histidine as targets. Free radicals were generated by copper and H2O2, or by gamma radiolysis. Protein-bound histidine was substantially converted into aspartate. Much proline was modified during radical attack, but it was not converted into glutamate. We conclude that histidine and proline are important sites of protein attack by radicals; protein cleavage may result from these reactions. PMID- 2572517 TI - Somatostatin inhibits the effect of secretin on bile flow and on hepatic bilirubin transport in the rat. AB - Increasing amounts of porcine secretion (0.05 to 2.00 clinical units/h/100 g body wt) given to rats during a continuous infusion of bilirubin, increased bile flow and the apparent maximal biliary excretion of bilirubin ('Tm'). This increment was caused by an enhanced biliary output of bilirubin monoconjugates. The effect was dose dependent but maximal at a secretin infusion of 0.80 CU. Somatostatin 0.2 and 0.8 microgram/h/100 g body wt caused a dose related inhibition of the hepatic effects of secretin both on bile flow and on biliary output of bilirubin conjugates. As secretin elicits the release of somatostatin, a feed-back system could be envisaged whereby the somatostatin released stops the effects of secretin. PMID- 2572518 TI - Rebound nocturnal hypersecretion after H2-receptor antagonist. PMID- 2572519 TI - [Reconstruction of the foot using free flaps]. AB - Microvascular free-flap reconstruction of the foot was performed during a seven year period in 25 patients to repair soft tissue loss caused mainly by traumatic injury. A scapular flap was used in fifteen cases, a dorsalis pedis in three, a radial forearm in two, a latissimus dorsi in two, and a gluteal thigh, rectus abdominis and tensor fascia lata each in one case. Two flaps were lost because of postoperative vascular complications. Soft-tissue stability was excellent or good in all of the seven patients with a successful free-flap transfer to the non weightbearing part of the foot and in eleven of the sixteen patients with a flap on the weightbearing part of the foot. Four patients had frequent superficial ulcerations of the graft but only one flap was completely unstable. Sixteen patients had normal or near normal ability to walk. Significant gait problems in seven patients were caused by skeletal deformity of the reconstructed foot or other associated injuries of the lower limb rather than the reconstruction itself. The best fit and contour was provided by thin skin flaps such as the dorsalis pedis and radial forearm flaps while the scapular flap offered a more inconspicuous donor site and could be used to cover large defects. Sensibility of the flaps assessed by clinical methods was less than normal in all cases. Good sensibility was found in the flaps of two children, fair in six, poor or very poor in fifteen, and two flaps had no sensibility. The level of sensibility was not related to the soft-tissue stability of the flap.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572520 TI - [Inborn errors of metabolism: lessons from a clinical case]. AB - Dramatic advances in recent years in the diagnosis and treatment of inborn errors of metabolism (IEM) make it imperative for the physician to be both aware of their occurrence and acquainted with their clinical presentations. Many may present with symptoms and signs common to sick infants, such as refusal to feed, vomiting and convulsions. Failure to include IEM in the differential diagnosis may have grave consequences, because only prompt recognition and appropriate treatment of a metabolic crisis can help prevent irreversible brain damage or death. We describe a 3-week-old female infant, eventually diagnosed as having an IEM (3-hydroxy-3-methyl-glutaric aciduria), who was admitted to various hospitals and died during her last admission. We focused on the clinical and laboratory findings of each of the first 2 admissions which might have provided clues to the true nature of the illness had the clinicians been aware of the possibility of IEM. Patients with IEM are generally managed in specialized centers; however, the responsibility for initial recognition and immediate treatment of a metabolic emergency lies with the primary physician. The purpose of our case presentation and the discussion of the lessons derived from it, is to prompt the clinician to an awareness and understanding of a subject that used to be considered reserved for specialists. In fact, all that is needed is clinical alertness and the performance of certain well-focused, simple laboratory tests for IEM. PMID- 2572521 TI - [Synergistic effect of irsogladine maleate and histamine H2-receptor antagonists on experimental gastric ulcers in rats]. AB - Effects of irsogladine maleate (IM), in combination with histamine H2-receptor antagonists or a muscarinic receptor antagonist, on the formation of stress ulcer were investigated in rats. The ED50 of IM and that of cimetidine for suppressing stress ulceration were remarkably reduced when these drugs were used in combination. ED50s in this case were less than the theoretical values calculated on the basis of additive action, thereby suggesting the synergistic effect of IM and cimetidine. The synergistic effect of IM and ranitidine or famotidine in suppressing stress ulceration was also observed, while IM and pirenzepine did not always produce a synergistic effect. In addition, for acetic acid-induced gastric ulcers, combined administration of IM and cimetidine also markedly potentiated ulcer healing. The marked synergistic potentiation of IM and histamine H2 receptor antagonists may be due to compensatory coordination of both drugs on the gastric secretion and mucosal microcirculation. These results suggest that the combination of IM and histamine H2-receptor antagonists may be beneficial in clinical gastric ulcer therapy. PMID- 2572522 TI - Split-thickness skin excision: its use for immediate wound care in crush injuries of the foot. AB - Split-thickness skin excision (STSE) was used as an adjunctive modality in the treatment of eight crush injuries of the foot. Compartment syndromes were present in four feet and were treated with fasciotomy. Wound debridement, internal fixation of fractures, and STSE followed. This technique accurately determined the viability of the skin flap, simultaneously providing skin for local wound coverage. All flaps treated in this manner survived and all (100%) of the degloved STSE grafts healed. Additional procedures were performed in four patients (two free flaps and two split-thickness skin grafts) adjacent to the debrided flap for complete coverage. STSE proved to be an effective modality for skin coverage in crush injuries of the foot associated with degloving of skin. PMID- 2572523 TI - Effect of islet-activating protein (IAP) on glucagon release from isolated perifused rat islets. AB - The present study was undertaken to evaluate the effect of islet-activating protein (IAP) on glucagon release using perifused isolated rat pancreatic islets. Glucagon release stimulated by 20 mM arginine was significantly enhanced in the IAP-treated rat pancreatic islets as compared with the IAP-untreated controls. Additionally, the effect of 1 ng/ml somatostatin on glucagon release was examined during ongoing stimulation by arginine. In the IAP-treated islets somatostatin inhibits glucagon secretion to the same extent as in the IAP-untreated islets. These results demonstrate that IAP potentiates arginine-induced glucagon release from perifused rat pancreatic islets, and IAP fails to antagonize the inhibitory effect of somatostatin in the pancreatic A cell. PMID- 2572525 TI - Multiple endocrine neoplasia type 2. International symposium. Heidelberg, November 5-7, 1987. PMID- 2572526 TI - Post-translation processing of neurohormonal peptide precursors in a human medullary thyroid carcinoma. AB - Determination of the Primary structures of peptides isolated from a human medullary thyroid carcinoma has provided insight into the pathways of post translational processing of prohormones in the tumour cells. Cleavage of the signal peptide in preprocalcitonin occurs at the Ala25-Ala26 bond. The prohormone is further processed at the site of multiple basic residues to generate procalcitonin-(1-57)-peptide, procalcitonin-(60-91)-peptide (calcitonin), procalcitonin(60-116)-peptide (a C-terminally extended form of calcitonin) and procalcitonin-(96-116)-peptide (Katacalcin). CGRP-I but not CGRP-II, was isolated from the tumour together with a high-Mr form that may represent the unprocessed prohormone. Progastrin-releasing peptide was processed to a mixture of GRP-(1-27) peptide and GRP-(18-27)-peptide, the latter component arising from proteolytic cleavage at the site of a single arginyl residue. PMID- 2572524 TI - Short tertatolol treatment does not impair the hormone and metabolic responses to exercise and hypoglycemia in diabetics. AB - Beta-blockers can precipitate hypoglycemia and mask its warning signs. Ten male insulin-dependent, otherwise healthy diabetic patients underwent two submaximal exercise tests and two insulin-induced hypoglycemic events (0.2 u/Kg short-acting insulin IV) after six days administration of placebo followed by tertatolol, a non selective beta-blocker (5 mg once daily). Tertatolol modified neither the exercise-induced changes in blood glucose, lactate and plasma unesterified fatty acid levels, nor those of counter regulatory hormones (glucagon, growth hormone, cortisol), while blood pressure, heart rate and plasma renin activity were significantly reduced, proving that tertatolol had actually been ingested, and was active. During the insulin-induced hypoglycemia, similarly tertatolol did not modify the course of the plasma fuels and hormones. Particularly, hypoglycemia was neither deeper nor more prolonged in the presence than in the absence of tertatolol. Warning symptoms were not affected except for palpitations which were not perceived. These results suggest that tertatolol did not precipitate hypoglycemia following exercise, and did not aggravate insulin-induced hypoglycemia in short term administration, and in otherwise healthy diabetic patients. PMID- 2572529 TI - The mapping of the locus for multiple endocrine neoplasia type 2A by linkage with chromosome 10 markers. AB - The sequence of studies leading to the assignment of the locus for multiple endocrine neoplasia (MEN2A) to chromosome 10 are described. They began with the exclusion of linkage between the disease locus and loci for conventional markers and a deletion site reported to be associated with the disease on chromosome 20. After 32% of the genome had been excluded, a "hint" of linkage was found but considerable additional family data were necessary in order to establish linkage to the marker locus D10S5. The DNA marker D10S5 was assigned to chromosome 10 by in situ hybridization and tests of linkage between the marker and disease loci were significant. The assignment of the marker and the linkage implied that the disease gene was on chromosome 10. The assignment was confirmed by the demonstration of an additional linkage, between the disease locus and the gene for interstitial retinol binding protein (RBP3) that had also been assigned to chromosome 10. Closer and flanking markers are now being sought as steps to providing better than Mendelian risks for the MEN2A genotype and for the ultimate identification of the gene. PMID- 2572530 TI - MEN-2 tumor associations suggest a linear order of specific endocrine tumor genes. AB - The variation in tumor involvement between families and consistent pattern within 12 MEN-2A and 4 MEN-2B families studied suggest that the tumors in MEN-2 result from separate, specific mutations in a specific linear order of adjacent genes. PMID- 2572527 TI - A young woman with pituitary-tumor, Cushing's syndrome and medullary thyroid carcinoma--a rare case of MEN II? AB - A young woman presented with Cushing's syndrome. As a repeat surgical resection of a pituitary tumor did not cure her a bilateral adrenalectomy was performed, which resulted in symptomatic improvement of the patients condition. Several months later, a medullary carcinoma of the thyroid (MTC) was detected and resected. Retrospective analysis suggests that a paraneoplastic Cushing's syndrome may have been present in this patient. A connection of the pituitary tumor and the MTC could not be proved, a MEN-II-syndrome does not seem likely. PMID- 2572528 TI - Pheochromocytoma-induced hypertensive encephalopathy revealing MEN-IIa syndrome in a 13-year old boy. Implications for screening procedures and surgery. AB - In the multiple endocrine neoplasia (MEN) type II syndrome, pheochromocytomas become manifest at a later age than medullary thyroid carcinomas (MTC) do. The present report concerns a 13-year-old boy, belonging to a MEN-IIA kindred, who was admitted because of convulsive seizures related to hypertensive encephalopathy. A pheochromocytoma was suspected immediately and appropriate medical therapy was initiated. A right adrenal pheochromocytoma was removed, as well as a pheochromocytoma of an accessory right adrenal gland. Today, three years later our patient is still asymptomatic and the results of the thyroid C cell provocative tests remain normal. This case clearly justifies the conclusion that periodic investigation of MEN-II family members to detect both medullary thyroid carcinoma and pheochromocytoma should begin early in life, as the latter may be the initial life-threatening expression of the disease. Long-term follow up of patients treated with unilateral adrenalectomy will permit better definition of the risk of contralateral recurrence in such cases. PMID- 2572531 TI - The expression of the neu oncogene product in breast lesions and in normal fetal and adult human tissues. AB - The expression of the neu oncogene product was investigated in invasive and non invasive ductal carcinomas of the breast, non-neoplastic lesions of the breast, fragments of normal adult and fetal breasts and in several other normal and fetal tissues at different weeks of pregnancy by means of an immunohistochemical study with monoclonal antibodies. The staining pattern along the cytoplasmic membrane was specific for malignancy and occurred in 29% of the breast carcinomas. It was observed in invasive carcinomas as well as in ductal carcinoma in situ and it showed a significantly higher expression in premenopausal women than in postmenopausal women. This higher expression was also present in oestrogen receptor-negative tumours. The tubules of the fetal and adult kidney, the absorption cells of the fetal and adult small and large intestine, the sebaceous glands of the fetal and adult small and large intestine, the sebaceous glands of the fetal and adult skin, the adult endocervix, the endometrium, the C-cells of the thyroid, hepatocytes and all ductal cells of the fetal breast showed a constant diffuse intracytoplasmic granular staining. staining. The same granular intracytoplasmic staining pattern was focally observed in rare cases of normal breast tissue in adults and in some cases of epitheliosis, aprocrine metaplasia and some breast carcinoma cells, which did not express neu oncogene product on their membrane. Western blot experiments showed that the cytoplasmic protein had a molecular weight of 155 kD (kilodaltons); the membrane protein is the known 185 kD neu protein. PMID- 2572532 TI - Neuroleptic dose reduction in persistently psychotic patients. PMID- 2572533 TI - Diagnosis of genetic disease using recombinant DNA. Second edition. AB - Recombinant DNA methodology has greatly increased our knowledge of the molecular pathology of the human genome at the same time as providing the means to diagnose inherited disease at the DNA level. Direct detection and analysis of a range of genetic defects are now possible using cloned gene or oligonucleotide probes or by direct sequencing of the disease gene(s). In addition, the use of restriction fragment length polymorphism (RFLPs) within and around these genes as indirect genetic markers has now potentiated the tracking of disease alleles in affected pedigrees in cases where direct analysis was not feasible. RFLPs associated with linked anonymous segments may also be used not only to diagnose hitherto undetectable disease states, but also for chromosomal localization of the loci responsible. We present here an up-to date list of reports describing both the direct and the indirect analysis/diagnosis of human inherited disease, which is intended to serve as a guide to current molecular genetic approaches in diagnostic medicine. PMID- 2572535 TI - Apolipoprotein (apo) E genotypes by polymerase chain reaction and allele-specific oligonucleotide probes: no detectable linkage disequilibrium between apo E and apo CII. AB - Allelic sequence variation in the apolipoprotein (apo) E gene has been analysed by means of synthetic oligonucleotide probes that detect single base pair substitutions in the codons for amino acid positions 112 and 158, substitutions that are responsible for the common isoforms. Use of the polymerase chain reaction procedure to amplify a sequence of 330 base pairs of the human apo E gene has permitted the development of a robust method for apo E genotyping. This technique has been used to determine the apo E genotype in 95 individuals in whom the genotype for an apo CII TaqI restriction fragment length polymorphism has also been determined. No strong linkage disequilibrium between the two gene loci was detected. This suggests that the metabolic effects of variation in the apo E and apo CII genes, as detected by the polymorphisms used here, would operate in a statistically independent manner. PMID- 2572534 TI - Linkage analysis of hereditary thyroid carcinoma with and without pheochromocytoma. AB - The use of polymorphic DNA segments as markers for the gene for the multiple endocrine neoplasia (MEN) syndrome, type 2a, allows the identification of family members at high risk for developing medullary carcinoma of the thyroid and other tumors, especially pheochromocytoma. Several families have also been identified in which medullary thyroid carcinoma is inherited, but pheochromocytoma is not seen. We have analysed 18 families, 9 with MEN 2A and 9 with medullary carcinoma of the thyroid without pheochromocytoma, with probes specific for the pericentromeric region of chromosome 10 and conclude that the mutations for the two presentations are closely situated. Genetic heterogeneity of the susceptibility locus was not seen among this sample of 18 families. The genetic mutation for medullary carcinoma was in disequilibrium with the marker alleles of the two closely linked probes, IRBPH4 and MCK2. These data suggest that different mutant alleles of the same gene or closely linked mutations account for the variation in penetrance of pheochromocytoma in families with hereditary medullary thyroid carcinoma. PMID- 2572536 TI - Haplotype frequencies of the collagen type-I genes in the Italian population. AB - We have determined the frequencies of six restriction fragment length polymorphisms (RFLPs) of type-I collagen genes in a random sample of 100 subjects. Alpha 1 gene (COL1A1) DNA polymorphisms, FG2/MspI, 2FC6/RsaI, and NST70/RsaI, had polymorphism information content (PIC) values of 0.35, 0.32, and 0.26, respectively. Alpha 2 gene (COL1A2) RFLPs, NJ3/EcoRI, Hf32/RsaI, and Hf32/MspI had PIC values of 0.36, 0.35, and 0.25, respectively. The combined haplotype PIC values were 0.71 at the COL1A1 locus and 0.73 for COL1A2. Two COL1A1 and two COL1A2 RFLPs were more polymorphic than in the English population, making them better markers for the analysis of Italian families affected by osteogenesis imperfecta and some other inherited collagen diseases. PMID- 2572537 TI - The beta subunit locus of the human fibronectin receptor: DNA restriction fragment length polymorphism and linkage mapping studies. AB - The beta subunit of the human fibronectin receptor (FNRB) is a transmembrane protein belonging to the VLA (very late antigens of activation) family. Using pGEM-32, a 2.5-kb partial cDNA clone corresponding to the 3' portion of the human FNRB locus, multiple restriction fragment length polymorphisms (RFLPs) were revealed on DNAs from unrelated Caucasians. RFLPs detected by five enzymes, BanII, HinfI, KpnI, BglII, and SacI, are of the simple two-allele form, and pairwise linkage analyses of these RFLPs with numerous known DNA markers from the chromosome-10 pericentromeric region not only confirmed the chromosome-10 assignment of the functional FNRB gene but also supported its localization at p11.2 suggested by in situ hybridization. An infrequent MspI RFLP was detected by pB/R2, a 4.6-kb genomic clone from the FNRB locus. Another type of DNA polymorphism was also revealed by the cDNA clone and it was visualized on the Southern blot analyses as the presence or absence of an extra band (or a set of extra bands). It seems to stem from a stretch of DNA sequence present in some individuals at one single locus but absent in others, and is of non-chromosome-10 origin based on linkage analyses with known chromosome 10 markers. This "presence/absence" type of polymorphism could be revealed by all of the 25 restriction enzymes tested and is similar in nature to that previously reported with one of the human dihydrofolate reductase pseudogenes, DHFRP1. Dissection of the pGEM-32 clone demonstrated that the region revealing the non-chromosome-10 sequences is within a fragment about 1.7 kb in length extending from about 600 nucleotides preceding the stop codon down to the end of the cloned FNRB 3' untranslated region. Due to its high polymorphism information content (PIC) value (0.71 for haplotypes of BanII, HinfI, and KpnI RFLPs) and proximity to the centromere. FNRB will prove to be a highly useful marker for genetic linkage studies of multiple endocrine neoplasia type 2A (MEN2A) as well as for chromosome 10 linkage studies in general. PMID- 2572538 TI - Linkage disequilibrium and CF allele segregation analysis in cystic fibrosis families in Northern Ireland. AB - Linkage disequilibrium and cystic fibrosis (CF) allele segregation were analysed in 46 CF families in Northern Ireland. The smaller (+) allele of the KM19/PstI polymorphism and the larger (-) allele of the XV-2c/TaqI polymorphism showed marked linkage disequilibrium with CF. This information can be used to alter the risk of an individual being a carrier of CF away from the expected population risk of 1 in 20. The high-risk genotypes K+K+ or X-X- have a risk of 1 in 10 and the low-risk genotypes K-K- or X+X+ have a risk of 1 in 50. A study of the segregation of CF alleles in the 46 families, using KM19 and Xv-2c, showed preferential inheritance of the paternal (79%), as opposed to the maternal (21%), CF allele by the heterozygous carriers. A mechanism that might explain this observation is discussed. PMID- 2572539 TI - TaqI polymorphism within the c-Ha-ras-1 VTR is associated with melanoma. PMID- 2572540 TI - A research agenda in care for patients with Alzheimers disease. PMID- 2572541 TI - The selective antigen-presenting cell capacity of activated B lymphocytes in HLA II-restricted responses of CD4+ T lymphocytes. AB - We examined the role of antigen-presenting B lymphocytes using panels of antigen specific CD4+8-T-lymphocyte clones (TLC). All 19 TLC showed a class II major histocompatibility complex-encoded (HLA-II) restricted proliferation to antigen presented by antigen-presenting cells (APC) from the monocyte fraction of peripheral blood. Only six TLC were effectively activated by antigen presented by autologous B lymphocytes activated by EBV transformation. This failure of B lymphocytes was not due to: (i) a high degree of cell surface sialic acid; (ii) a low expression of the cell surface proteins HLA-II, ICAM-1 or LFA-3 that restrict antigen presentation; (iii) lack of secretion of the cytokine IL-1 or other soluble factors that may be required as secondary signals; or (iv) induction of incomplete T-cell activation resulting in the production of growth factor interleukin-2 (IL-2) or the expression of receptors for IL-2 only. These data suggest the involvement of another cell surface interaction in antigen presentation acting besides the interactions known so far. PMID- 2572542 TI - [Report of a case of allergic granulomatous vasculitis (Churg-Strauss syndrome)]. AB - A 35 year female with a previous history of asthma came to our observation about 10 months ago. She was dyspneic, showed small nodules (2 to 5 mm) in arms, hands and legs and had radiological evidence of pulmonary granulomatosis and pleural effusion. Diagnosis was made by means of cutaneous biopsy showing a necrotic granulomatous vasculitic lesions with eosinophilic infiltration and giant cells, consistent with allergic granulomatosis of Churg-Strauss syndrome. Steroid therapy alone (prednisone 1 mg/kg/die) led to a rapid and complete clinical and humoral remission. The patient is still doing an alternate day prednisone therapy (15 mg) and is well after a 10 months follow-up. PMID- 2572543 TI - [Preliminary approach to the mental component in dermatologic patients]. AB - The aim of this study was to approach on a psychical point of view 27 patients suffering from chronic idiopathic urticaria (14), pruritus sine materia (6), alopecia areata (3), pathomimia (4), in order to examine the possibility that psychic disorders could act as triggering or aggravating the dermatological affection. Psychical assessment was evaluated by colloquy and by the administration of some psychodiagnostic tests: EPI, MMPI, Zung. The role of psychogenic factors in skin diseases is emphasized and the results obtained from the use of antidepressant and minor tranquilizer drugs are discussed. PMID- 2572544 TI - Expression of a new human THY-1 related antigen in Ewing's sarcoma and peripheral neuroectodermal tumors. AB - Ewing's sarcoma (ES), peripheral neuroectodermal tumor (PNET) and Askin's tumor of the chest wall share a reciprocal chromosomal translocation between the long arms of chromosomes 11 and 22 (q23-24; q12). In the absence of other distinguishing features this specific translocation is regarded as marker of a common and neuroectodermal origin for these rare tumors. A monoclonal antibody (HBA-71) developed in our laboratory has been found to recognize an unique ES and PNET associated antigen, which is also expressed in some normal tissues, including thymus, bone marrow, islets of Langerhans, ependyma and adenohypophysis. It is shown in this study that this HBA-71 antigen is closely related to the murine THY-1 antigens, major cell surface glycoproteins of thymocytes and brain in mice and rat. Both antigens have similar molecular ratios (18,000), amino acid compositions and sensitivity to tryptic digestion, show high cell surface expression, and binding of the appropriate antibodies to HBA-71 antigen triggers proliferation in thymocytes. The HBA-71 epitope may represent a primitive neuroectodermal marker of ES/PNET, or its expression may be directly linked to the reciprocal translocation invariably associated with HBA-71-positive ES and PNET tumors, which maps to the same region of chromosome 11 (q23-24) as the human Thy-1 gene. PMID- 2572545 TI - Effect of beta-adrenoceptor antagonists and some related drugs on maximal electroshock seizures in mice. AB - (+/-) Propranolol (1-50 mg/kg), (+) propranolol (50 mg/kg) and pindolol (10-50 mg/kg) exhibited significant protective effects against MES (maximum electroshock seizures), whereas, timolol (1 mg/kg), the propranolol analog, UM-272 (1 and 10 mg/kg), and the beta-agonist, terbutaline (1 and 10 mg/kg) were ineffective. Cholinergic agents, physostigmine (0.01-1.0 mg/kg), and atropine (1 and 10 mg/kg), the serotonin antagonist, cyproheptadine (0.05 mg/kg), and the prostaglandin synthesis inhibitor, indomethacin (10 mg/kg), were also without effect on the MES extensor phase. Further, pretreatment of mice with terbutaline, atropine, cyproheptadine or indomethacin did not influence the anti-MES effect of propranolol to any significant extent. The results indicate that the observed anticonvulsant effects of beta-adrenoceptor antagonists are unrelated to noradrenergic or other central neurotransmitter systems and that a non-specific mechanism, probably a membrane stabilizing effect is involved. PMID- 2572546 TI - Effect of increased level of CO2 exposure in a closed environment on calcium and phosphorus balance in rats. AB - In rats, exposed to increased ambient CO2 level (1.5%) for 30 days a 40% reduction in food intake during first 10 days was observed which subsequently improved. Similarly the body weight also declined initially, which showed progressive gain thereafter, almost reaching that of ad libitum fed control by 30th day. The lowered Ca intake consequent to reduced food consumption was followed by diminution in renal and faecal excretion of Ca. The lowered renal Ca excretion was probably unrelated to reduced Ca intake, since the pair-fed control, not exposed to raised CO2 level, did not show any such alteration. The net result was a drastic reduction in Ca balance despite the diminution in its excretion. Though the intake and renal excretion of P were reduced in the CO2 exposed group due to a reduction in food intake, the P balance showed a cyclic pattern as in the pair-fed controls. The serum Ca after 30 days' exposure remained unchanged while the serum inorganic P showed a variation. The data indicated that hypercapnia produced hypophagia which affected growth of rats. The reduced renal excretion of Ca may not pose a risk on calcification of soft tissues, since its retention was also reduced on exposure to raised concentration of CO2. PMID- 2572547 TI - Deletion, insertion, and restriction site polymorphism of the T-cell receptor gamma variable locus in French, Lebanese, Tunisian, and black African populations. AB - The human T-cell receptor gamma region spans 160 kb of genomic DNA and is densely populated by coding sequences. Restriction fragment length polymorphisms have been previously documented for the constant region genes, the joining segments, and the variable genes belonging to subgroups I and IV. Here we further define the polymorphism of the V gamma I subgroup genes. based on complete mapping of the Eco RI and Taq I allelic restriction fragments. We describe seven haplotypes; five result from polymorphic restriction sites, the sixth corresponds to a deletion of about 10 kb encompassing V4 and V5, and the seventh results from an insertion of an additional gene, V3P, between V3 and V4. As a consequence of the deletion or insertion polymorphism, the number of V gamma I subgroup genes vary from seven in haplotype TRGVI*3 to ten in haplotype TRGVI*4, whereas the most common haplotype, TRGVI*1, has nine V genes, five of them being functional. Frequencies of the different TRGVI haplotypes in French, Lebanese, Tunisian, and Black African populations are given. PMID- 2572548 TI - Restriction fragment analysis of H-2 recombinant mouse strains with crossovers between E alpha and C4 genes. PMID- 2572549 TI - Comparative analysis of the structures of the outer membrane protein P1 genes from major clones of Haemophilus influenzae type b. AB - P1 outer membrane proteins from Haemophilus influenzae type b are heterogeneous antigenically and with respect to apparent molecular weight in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. For determination of the molecular basis for the differences in the P1 proteins, the genes for the P1 proteins from strain 1613, representative of outer membrane protein subtype 3L, and strain 8358, representative of outer membrane protein subtype 6U, were cloned, sequenced, and compared with the previously reported gene for the P1 protein from strain MinnA, a strain with the outer membrane protein subtype 1H. These prototype strains are representatives of the three major clonal families of H. influenzae type b responsible for invasive disease in diverse areas of the world. The nucleotide sequences of the P1 genes from strains 1613 and 8358 were 94 and 90% identical to the MinnA sequence, respectively. The derived amino acid sequences were 91 and 86% identical, respectively. Heterogeneity between the MinnA and 1613 proteins was largely localized to two short variable regions; the protein from strain 8538 contained a third variable region not observed in the other P1 proteins. Thus, the outer membrane protein P1 genes are highly conserved; the variable regions may code for the previously demonstrated strain specific antigenic determinants. PMID- 2572550 TI - Adhesion and ultrastructural properties of human enterotoxigenic Escherichia coli producing colonization factor antigens III and IV. AB - Human enterotoxigenic Escherichia coli (ETEC) producing colonization factor antigen III (CFA/III) and coli surface antigens 4, 5, and 6 (CS4, CS5, and CS6) of CFA/IV were examined ultrastructurally and for ability to adhere to human small intestinal enterocytes and to cultured human intestinal mucosa. Strains of serotypes O25:H-, O25:H42, and O167:H5 producing CFA/III plus CS6, CS4 plus CS6, and CS5 plus CS6, respectively, showed good adhesion to human enterocytes (1.8 to 4.2 bacteria per brush border) and cultured human intestinal mucosa, whereas variants lacking these antigens or producing only CS6 were nonadherent (0 to 0.03 bacterium per brush border). By electron microscopy, CFA/III, CS4, and CS5 appeared as morphologically distinct rodlike fimbriae: CFA/III was 7 to 8 nm in diameter, CS4 was 6 to 7 nm in diameter, and CS5 was 5 to 6 nm in diameter. CS5 was unusual in that it appeared to be composed of two fine fibrils arranged in a double-helical structure. CS6 was difficult to characterize morphologically but possibly has a very fine fibrillar structure. By specific fimbrial staining and immunoelectron microscopy. CS4 and CS5 were shown to promote mucosal adhesion of ETEC; a similar adhesion role for the CS6 antigen could not be confirmed. ETEC strains of serotypes O27:H7, O27:H20, O148:H28, and O159:H20 which produced CS6 showed good adhesion to human enterocytes (1.6 to 3.0 bacteria per brush border), whereas variants which lacked CS6 were nonadherent (0 to 0.01 bacterium per brush border). These strains, however, also produced fimbrial or fibrillar surface antigens, in addition to CS6, which probably represent additional coli surface antigens responsible for the observed adhesive properties of these ETEC serotypes. PMID- 2572551 TI - Binding specificities of wild-type and cloned Escherichia coli strains that recognize globo-A. AB - In this study we compared the specificity for the globoseries of glycolipids of Escherichia coli expressing the O-negative, A-positive (ONAP) adhesin and clones transformed with the pap-like (prs or pap-2) gene cluster. Receptor-active glycolipids were identified by the ability of radiolabeled bacteria to bind to the glycolipids on thin-layer chromatogram plates. The ONAP adhesin and pap-like clones bound with high affinity to the globo-A and Forssman glycolipids. The ONAP strains did not recognize other glycolipids of the globoseries. In contrast, the pap-like clones also showed weak binding to globotriaosylceramide and reacted weakly with Gal alpha 1----4 Gal beta-latex beads. We suggest that the pap-like and ONAP adhesins recognize an epitope shared by the globo-A and Forssman structures, e.g., terminal GalNAc alpha 1----3 bound to Gal alpha 1----4Gal beta containing glycolipids. PMID- 2572552 TI - Use of adhesin-specific monoclonal antibodies to identify and localize an adhesin on the surface of Capnocytophaga gingivalis DR2001. AB - Monoclonal antibodies capable of inhibiting coaggregation between Capnocytophaga gingivalis DR2001 and Actinomyces israelii PK16 were used to identify the adhesin on C. gingivalis that mediates the interaction. The monoclonal antibodies were used to demonstrate that a 140-kilodalton polypeptide found in the outer membrane of C. gingivalis was the adhesin responsible for coaggregation. A coaggregation defective mutant that was unable to coaggregate with A. israelii lacked this large polypeptide. The monoclonal antibodies were also used to estimate the number of binding sites on the surfaces of individual cells and show how the adhesin molecules were arranged on the outer membrane. Values of between 220 and 280 were obtained for the number of adhesin molecules per cell. Immunoelectron microscopy performed with the monoclonal antibodies revealed that the adhesin molecules were arranged nonuniformly on the bacterial surface and occurred singly, in pairs, and in small clusters. PMID- 2572553 TI - Production and characterization of monoclonal antibodies to a pilus colonization factor (colonization factor antigen III) of human enterotoxigenic Escherichia coli. AB - Three monoclonal antibodies (MAbs) to a pilus colonization factor (colonization factor antigen III [CFA/III]) of human enterotoxigenic Escherichia coli (ETEC) were developed and characterized. All of the MAbs isolated belonged to the immunoglobulin G2a subclass. The specificity of these MAbs for CFA/III pili was demonstrated by the immunogold-labeling technique. The presence of more than one epitope in CFA/III pili was suggested. One of the three MAbs appears to recognize a polymeric conformational epitope(s) of CFA/III. CFA/III antigenicity distinct from that of other pilus colonization factors of ETEC was demonstrated by both a bacterial agglutination test and a sandwich enzyme-linked immunosorbent assay using the MAbs. Of the 100 strains of ETEC isolated from persons with traveler's diarrhea, 8% were found to carry CFA/III pili. Two enzyme-linked immunosorbent assay systems which could detect as little as several or 50 ng of CFA/III per ml were developed. PMID- 2572554 TI - Mucosal and systemic immune responses in BALB/c mice to Bacteroides gingivalis fimbriae administered orally. AB - A 41,000-molecular-weight fimbrial protein was isolated from freshly cultivated whole cells of Bacteroides gingivalis 381 and purified chromatographically. Salivary and serum antibody responses to the fimbriae, which had been orally administered in the presence of an acyl derivative of muramylpeptides, i.e., either N2-[(N-acetylmuramyl)-L-alanyl-D-isoglutaminyl]-N6-stearoyl-L-lysine [MDP Lys(L18)] or sodium beta-N-acetyl-glucosaminyl-(1----4)-N-acetylmuramyl-L-alanyl D-isoglu tam inyl- (L)-stearoyl-(D)-meso-2,6-diaminopimelic acid-(D)-amine-D alanine (GM-53), or in the absence of adjuvant, were examined in BALB/c mice when administered by gastric intubation on days 0 and 1 as primary immunizations and on days 27 and 28 as booster immunizations. Gastric intubation of the fimbriae with an adjuvant significantly enhanced the production of anti-fimbria immunoglobulin A (IgA) in saliva. Subcutaneous injection of fimbriae along with an adjuvant also raised anti-fimbria IgA levels, as well as IgG levels, in saliva. Both immunization procedures enhanced the levels of anti-fimbria IgG, IgA, and IgM in serum, and the major class of fimbria-specific antibody was IgG, followed by IgA and IgM. However, subcutaneous injection was more effective than gastric intubation to enhance the production of serum antibody in mice. The subclasses of IgG antibody specific for fimbriae in serum were mainly IgG1, followed by IgG2a, IgG2b, and IgG3. These results demonstrated that the combined use of B. gingivalis fimbrial antigen and either GM-53 or MDP-Lys(L18) resulted in a sharply increased IgA antibody response in saliva and a predominantly stimulated IgG antibody response in serum, respectively. Both antibodies were found to be specific for the fimbriae used for immunization. PMID- 2572555 TI - Nucleotide sequence analysis of a type 1 fimbrial gene of Streptococcus sanguis FW213. AB - A structural gene for type 1 fimbriae of Streptococcus sanguis FW213 was located within a 6-kilobase fragment cloned in Escherichia coli. A 1.6-kilobase internal fragment contains an open reading frame of 927 bases coding for an immunoreactive peptide of 34,349 daltons, which corresponds in size with an observed cytoplasmic form of fimbrial peptide (P. M. Fives-Taylor, F. L. Macrina, T. J. Pritchard, and S. J. Peene, Infect. Immun. 55:123-128, 1987). Disruption of the reading frame by insertional mutagenesis results in loss of immunoreactivity. Consensus sequences for initiation of transcription and translation were identified 5' to the coding region. Transcription terminator-like sequences were found downstream of the coding region. The deduced amino acid sequence of the cloned fimbrial peptide shows a strongly hydrophobic signal sequence at the amino terminus. The carboxyl terminal region does not include a hydrophobic membrane anchor sequence such as has been reported for other gram-positive surface structures. A hydrophobic region of 12 to 14 amino acids downstream from the putative signal sequence cleavage site exhibits homology with the Streptococcus pyogenes type 6 M protein repetitive region A (S. K. Hollingshead, V. A. Fischetti, and J. R. Scott, J. Biol. Chem., 261:1677-1686, 1986). Functional homology at the level of protein secondary structure with Actinomyces viscosus T14V type 1 fimbriae (M. K. Yeung, B. M. Chassy, and J. O. Cisar, J. Bacteriol., 169:1678-1683, 1987) is proposed. PMID- 2572556 TI - Role of L3T4+ lymphocytes in protective immunity to systemic Candida albicans infection in mice. AB - Protective immunity to lethal Candida albicans challenge in vivo and activation of splenic macrophages with highly candidacidal activity in vitro were detected in mice infected with low-virulence agerminative yeast cells of the variant strain PCA-2, at a time when a strong delayed-type hypersensitivity (DTH) reaction to C. albicans occurred in the footpads of PCA-2-treated mice. The DTH reaction was transferable with spleen cell populations from these animals, and enrichment of splenic lymphocytes in L3T4+ cells significantly increased the footpad swelling. The reactivity transferred by L3T4+ cells was a radiosensitive (2,500 rads in vitro) phenomenon that required collaboration with radioresistant, silica-sensitive syngeneic cells in the host and was inhibited by treatment of recipient mice with antibodies to the L3T4 antigen or murine gamma interferon. In vitro, the PCA-2-immune L3T4+ cells produced various lymphokine activities upon incubation with C. albicans, including gamma interferon and granulocyte macrophage colony-stimulating factor. Anti-L3T4 monoclonal antibody treatment of PCA-2-infected mice significantly impaired their footpad reaction and resistance to C. albicans, as shown by increased recovery of yeast cells from the kidneys of anti-L3T4-treated mice. These results suggested that the mechanisms of anti Candida resistance induced by PCA-2 may involve specific induction of a DTH response mediated by inflammatory L3T4+ T cells and lymphokine-activated phagocytic effectors. However, the survival rate of the PCA-2-immune mice challenged with C. albicans was not significantly modified by administration of the anti-L3T4 antibody, thus allowing for the conclusion that compensatory mechanisms lead to considerable anti-Candida resistance when the activity of L3T4+ cells is deficient. PMID- 2572557 TI - Purified P fimbriae from two cloned gene clusters of a single pyelonephritogenic strain adhere to unique structures in the human kidney. AB - We have completed immunofluorescence binding studies of purified fimbriae from two clones which express Pap or Pap-2 fimbriae. Although the two fimbrial types exhibited common binding to the uroepithelia of the bladder and renal pelvis and to occasional cells located within the glomeruli which we have termed glomerular elements, only Pap-2 fimbriae adhered to Bowman's capsule. Previous studies have demonstrated that the Gal alpha 1----4Gal disaccharide moiety is capable of inhibiting Pap hemagglutination and adherence to uroepithelial cells. Results of our experiments demonstrate that this disaccharide is not sufficient for blocking binding of Pap-2 fimbriae to Bowman's capsule but that GalNAc beta 1----3Gal completely blocks Pap-2 adherence to Bowman's capsule. These results indicate that the different hemagglutination capacities of the two clones reflect different receptor specificities and differential tissue tropisms in the urinary tract. These unique receptor specificities may provide uropathogenic strains of Escherichia coli carrying multiple chromosomal copies of pap-like gene clusters with the advantage of increased numbers of binding sites within the urinary tract. This, in turn, might improve the chances of colonization and the establishment of infection. PMID- 2572558 TI - Recombinant human gamma interferon enhances in vitro activation of lymphocytes isolated from patients with acquired immunodeficiency syndrome. AB - Decreased responses to antigens and lectins are a characteristic feature of peripheral blood lymphocytes isolated from patients with the acquired immunodeficiency syndrome (AIDS). The in vitro addition of recombinant gamma interferon (IFN-gamma) to cultures of peripheral blood lymphocytes obtained from patients with AIDS resulted in an augmented proliferative response [( 3H]thymidine uptake) to phytohemagglutinin (PHA) and an enrichment in CD4+ and CD8+ T lymphocytes. In AIDS cultures stimulated with PHA, IFN-gamma increased the release of T-cell growth factors and enhanced the expression of interleukin-2 receptors on activated lymphocytes. Responsiveness to PHA was augmented, albeit to a lesser extent, by IFN-gamma in cultures derived from normal donors. Proliferation to microbial antigens including herpes simplex virus, cytomegalo virus, and Candida albicans was increased by IFN-gamma in cultures established from a group of AIDS patients with Kaposi's sarcoma who had no history of opportunistic infection. PMID- 2572559 TI - Inhibition of human lymphoproliferative responses by mycobacterial phenolic glycolipids. AB - The effect of mycobacterial phenolic glycolipids from Mycobacterium leprae, M. bovis BCG, and M. kansasii on in vitro proliferative responses by human blood mononuclear cells from healthy BCG vaccinees was investigated. All three phenolic glycolipids inhibited proliferation in a concentration-dependent manner. Inhibition was independent of the stimulus used and involved neither antigen presenting cells nor antigen-specific CD8+ suppressor T cells. It was concluded that the phenomenon may be a general property of mycobacterial phenolic glycolipids, perhaps analogous to the growth-modulating properties of gangliosides. Despite the lack of specificity of inhibition in vitro, de facto specificity may occur in vivo by virtue of the localization of glycolipid in the leprosy lesions. PMID- 2572560 TI - Characterization of the Pseudomonas aeruginosa pilus adhesin: confirmation that the pilin structural protein subunit contains a human epithelial cell-binding domain. AB - Previous studies have suggested that the Pseudomonas aeruginosa PAK pilus adhesin moiety resides in an epithelial cell-binding domain located in the C-terminal region of the PAK pilin structural protein. Synthetic peptides Ac17red (a synthetic peptide with a sequence identical to that of PAK pilin residues 128 to 144, with the Cys-129 and Cys-142 residues being in the reduced state) and Ac17ox (a synthetic peptide with a sequence identical to that of PAK pilin residues 128 to 144, with a formed disulfide bridge between the amino acid residues Cys-129 and Cys-142), which should contain the epithelial cell-binding domain, were synthesized. Ac17red and Ac17ox both bound to buccal epithelial cells (BECs) and to ciliated tracheal epithelial cells (TECs). Ac17ox had a Km of 6.40 microM for binding to BECs, while Ac17red had a Km of 9.87 microM. Ac17red bound to the same receptor sites that purified pili did and competitively inhibited the binding of purified PAK pili to BECs. BEC glycoproteins with molecular masses of 82, 55 to 51, and 40 kilodaltons immobilized on nitrocellulose exhibited periodate sensitive receptor activity for Ac17red; similar activity has been found for PAK pili. Ac17red, Ac17ox, and PAK pili bound to the cilia and luminal portions of the cytoplasmic membrane of human TECs, the same regions to which P. aeruginosa whole cells bind. PAK pilin has an epithelial cell-binding domain that resides in the C-terminal region of the protein. PMID- 2572561 TI - Lethal infection by Bordetella pertussis mutants in the infant mouse model. AB - Different aspects of lethal infection of infant mice with Bordetella pertussis were examined. Mutants deficient in vir-regulated genes were tested for the ability to cause a lethal infection in the infant mouse model. Adenylate cyclase toxin-hemolysin and pertussis toxin were required to cause a lethal infection at low doses. Mixed infection caused by challenging the mice with an equal number of pertussis toxin and adenylate cyclase toxin-hemolysin mutants at a dose at which neither alone was lethal was also unable to cause a lethal infection. Production of the filamentous hemagglutinin and the dermonecrotic toxin was not required to cause a lethal infection. Nine other mutants in vir-regulated genes whose phenotypes have yet to be determined were also tested. Only two of these mutants were impaired in the ability to cause a lethal infection. Expression of fimbriae does not appear to affect the dose required to cause a lethal infection; however, fimbrial expression was correlated with the later stages of a nonlethal, persistent infection. Growth of the bacteria in MgSO4, a condition which reversibly suppresses expression of the genes required for virulence, did not alter the ability of the bacteria to cause a lethal infection. Auxotrophic mutants deficient in leucine biosynthesis were as virulent as the parental strain; however, mutants deficient in methionine biosynthesis were less virulent. A B. parapertussis strain was much less effective in promoting a lethal infection than any of the wild-type B. pertussis strains examined. A persistent infection in the lungs was observed for weeks after challenge for mice given a sublethal dose of B. pertussis, and transmission from infected infants to the mother was never observed. PMID- 2572563 TI - Bacteroides loeschei PK1295 cells express two distinct adhesins simultaneously. AB - Bacteroides loeschei synthesizes two distinct adhesins that mediate its coaggregation with Streptococcus sanguis 34 and Actinomyces israeli PK14. Streptococcal adhesin-specific and actinomyces adhesin-specific monoclonal antibodies were used to prepare antibody-coated 5- or 10-nm gold particles. These were used in immunoelectron microscopic studies to establish that essentially all bacteroides cells in a population express both adhesins. In general, the two sizes of gold particles representing each type of adhesin appeared to be spatially separated on neighboring fimbriae of B. loeschei. Deposition of antibody-coated gold particles, representing both types of adhesin, at or near the same fimbria was observed less frequently. PMID- 2572564 TI - [Perioperative glucose resorption and hormonal reaction following intraduodenal glucose administration]. AB - Early postoperative enteral nutrition has repeatedly been described in the literature but has not found its place yet in everyday clinical life. We conducted perioperative intraduodenal glucose tolerance tests in 12 patients with a healthy metabolism who had to undergo moderately severe abdominal surgery. Our results suggest that the resorption is still markedly delayed 12 h postoperatively and that the hormonal regulation is also essentially disturbed. Therefore, we believe that enteral nutrition in the early postoperative period is beneficial only more than 24 h after surgery. PMID- 2572562 TI - Pseudomonas aeruginosa pili as ligands for nonopsonic phagocytosis by fibronectin stimulated macrophages. AB - Fibronectin is capable of activating macrophages for enhanced nonopsonic phagocytosis of Pseudomonas aeruginosa grown in vivo in rats or mice or in vitro on nutrient agar plates. In this study it was determined that while fibronectin was able to significantly increase phagocytosis of organisms grown in static broth, uptake of agitated bacteria could not be promoted. Agitated P. aeruginosa cultures were proven to lack surface pili expression, as assessed by electron microscopic studies. A pilus-deficient pilA::Tn501 mutant of P. aeruginosa PAO was constructed by gene replacement techniques. Phagocytosis of this mutant could not be enhanced by fibronectin regardless of growth conditions. Furthermore, 60 micrograms of exogenously added Pseudomonas pili per ml was capable of abrogating the enhanced phagocytosis of the wild-type strain observed with fibronectin stimulated macrophages. It is concluded that Pseudomonas pili were the bacterial ligands required for attachment to fibronectin-stimulated macrophages in the initial stages of nonopsonic phagocytosis. PMID- 2572565 TI - Gamma-glutamyl transpeptidase in rat epididymis: effects of castration, hemicastration and efferent duct ligation. AB - Gamma-Glutamyl transpeptidase (gamma-GT) was studied histochemically and biochemically in the rat epididymis after castration with or without testosterone treatment, or after hemicastration and ligation of the efferent ducts. There was a strong reaction to gamma-GT in the apical part of the epithelium in the caput epididymis, while in the corpus and cauda the reaction was confined mainly to the luminal contents. Castration caused a marked decline in epithelial gamma-GT activity within 10 days. Subsequent testosterone treatment (1 mg/day for 10 days) restored gamma-GT activity in the apical surface and lumen. After hemicastration of adult rats, and after hemicastration or ligation of the efferent ducts in immature 28-day-old rats, a small but significant (P less than 0.001) decrease was observed in gamma-GT activity in the epididymal caput compared to controls. The quantities of six other enzymes (beta-N-acetylglucosaminidase, beta galactosidase, angiotensin-converting enzyme, alanyl amino-peptidase, dipeptidyl peptidase IV, acid phosphatase) also displayed significant changes after castration and restoration of activities by testosterone treatment. However, their distribution in the caput and cauda epididymis was more even than that of gamma-GT, and the changes after castration were less drastic. It is concluded that gamma-GT is a highly sensitive androgen-dependent secretory marker in the caput epididymis and may have an important function in sperm maturation. PMID- 2572566 TI - Comparison of once and twice daily oral dosing schedules of bevantolol, a new beta-adrenoceptor antagonist with alpha-adrenoceptor partial antagonist activity in patients with angina of effort. AB - In a placebo controlled double-blind cross-over study following a dose titration phase, we compared the efficacy of benantolol, a new beta 1 adrenoceptor antagonist with alpha-adrenoceptor partial antagonist activity at 12 and 24 hours after dosing in patients with angina of effort. Twenty patients aged 43-65 years were studied. Each study phase lasted four weeks. Efficacy was determined by treadmill exercise testing using the standard Bruce protocol at the end of each phase. Fifteen patients satisfactorily completed the study. Data from five protocol violators were not analysed. In the four patients who received bevantolol 200 mg daily, exercise time increased from 395 +/- 192 (mean +/- 1 SD) sec on placebo to 468 +/- 171 sec at 10-12 hours and to 442 +/- 230 sec at 22-24 hours after dosing with bevantolol. In the eleven patients who received bevantolol 400 mg daily, exercise tolerance of 290 +/- 103 sec on placebo increased to 408 +/- 112 sec at 10-12 hours (P = 0.001) and to 400 +/- 98 sec at 22-24 hours (P = 0.001) after dosing with bevantolol. Maximum exercise capacity at 10-12 and 22-24 hours after dosing with bevantolol were comparable. Maximum exercise heart rate and systolic blood pressure on placebo and on bevantolol at 10-12 and at 22-24 hours after dosing were comparable. Thus, bevantolol has salutary effects on exertional angina up to 24 hours after dosing. PMID- 2572567 TI - The v-erbA oncogene is a thyroid hormone receptor antagonist. PMID- 2572568 TI - Novel subsets of human T cells (CD4+ CD8- TCR gamma delta and CD4- CD8- TCR alpha beta) and T-cell development. PMID- 2572569 TI - Treatment for chronic CHD. AB - It seems likely that a number of medical and surgical treatments have had an impact on the overall mortality from ischaemic heart disease in recent years although there is comparatively little data upon which to base conclusions. The two forms of treatment which have been shown, in controlled trials, to reduce mortality are beta blockade after infarction and coronary artery bypass surgery in chronic ischaemic heart disease. It is of interest that these treatments seem to be most often applied in countries that have had the greatest fall in mortality, although it is not suggested that they have played more than a small role. It is likely that other influences on mortality have included the use of pacemakers and the avoidance of previously undesirable medical practices. Much more careful documentation of the use of treatments in different countries is highly desirable. PMID- 2572570 TI - Recognition and differential diagnosis of tardive dyskinesia. AB - Tardive dyskinesia (TD) is a consequence of chronic neuroleptic therapy. It is an irregular stereotyped movement disorder that is usually choreic in appearance, and is subject to temporary volitional control. Dystonia, akathisia, and tics are uncommon variants of the classic tardive syndrome. Characteristic clinical features including amelioration by action, augementation by distraction, partial volitional suppressibility, and lack of subjective distress help differentiate TD from other movement disorders such as resting tremor, Huntington's disease, spontaneous dyskinesias, and abnormal movements accompanying psychiatric illnesses. PMID- 2572571 TI - Differential susceptibility of isolated mouse retinal neurons and photoreceptors to kainic acid toxicity. In vitro studies. AB - Dissociated cultures of mouse retinal neurons and photoreceptors in chemically defined medium were used to investigate the susceptibility of these cells to the neurotoxin kainic acid (KA). Cells isolated from the newborn mouse retina were initially insensitive to this toxin, and the cells that differentiated as rod photoreceptors retained this resistance throughout the culture period. However, amacrine neurons became increasingly sensitive to KA toxicity as they differentiated in culture; after the fifth day in vitro approximately 90% of these cells were killed by KA in a concentration- and time-dependent fashion. The neurons showed pronounced swelling within 10 min of treatment onset, and cell lysis and nuclear fragmentation were evident during the next few hours. KA induced degeneration of these neurons was corroborated using the amacrine cell specific monoclonal antibody HPC-1, as well as autoradiographic and biochemical determinations of the high affinity uptake for GABA. This inhibitory neurotransmitter was taken up by amacrine neurons but not by photoreceptor cells, and this uptake was completely abolished in KA-treated cultures. Similar results were obtained with the neuronal enzyme choline acetyltransferase. However, both photoreceptor survival and the expression of photoreceptor markers such as opsin and the retinoid-binding protein interphotoreceptor (IRBP) were similar in KA treated and control cultures. Similarly, the high affinity uptake of glutamate, an excitatory amino acid which is predominantly taken up by photoreceptors, showed only modest changes in KA-treated cultures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572572 TI - [Unusual cutaneous angiofibromatosis following gold therapy of primary chronic polyarthritis]. AB - An unusual case of late cutaneous intolerance reaction following gold therapy for rheumatoid arthritis is reported. In keloidlike angiofibromatoid lesions that were spread over the upper part of the trunk and the arms, perivascular macrophages carrying gold deposits inside lysosomes were detected with two recently introduced methods--energy-dispersive X-ray microanalysis (EDX) and electron spectroscopic imaging (ESI)--for definitive ultrastructural localization. PMID- 2572573 TI - Proliferative activity of gastric and duodenal endocrine cells in the rat. AB - The replicative activity and migration of gastrin, somatostatin and serotonin cells in rat stomach and duodenum was studied using combined immunocytochemistry and autoradiography after 3H thymidine pulse-labeling. Our results show that a small proportion of gastrin, somatostatin and serotonin immunoreactive cells displays proliferative activity. The overall labeling index ranged from 1.3% for gastric endocrine cells to 3.2% for duodenal endocrine cells. In a pulse chase experiment, labeling indices of immunoreactive cells were estimated at several time intervals after 3H thymidine administration. Significant differences in labeling index were not found. Migration of 3H thymidine labeled endocrine cells towards the luminal surface was not found in the stomach nor in the duodenum. It is concluded that 1) these endocrine cells have replicating activity; 2) the replicative activity of endocrine cells is higher in the duodenum than in the stomach; 3) the various cell types do not show significant differences in replicating activity and 4) endocrine cells did not seem to migrate to the luminal surface of the mucosa along with the other epithelial cells. PMID- 2572575 TI - Assessment of mental workload on older workers during performance of a calculating task. AB - Twenty-four healthy male clerical workers (50-59 years of age) in an automobile manufacturing plant performed as accurately as they could a calculating task consisting of the addition or subtraction of two 2-digit numbers at five grades of task load for 5 min each. The workers' work capacities were inferior in speed but not in precision to those of male students determined previously. The resting levels of occipital midline beta-2 (Oz beta 2) amplitude were significantly lower for the workers than for the students. The workers' mental workloads were objectively shown to be lighter than the students' based on a significant difference between the percent increases in Oz beta 2 amplitude over resting levels. The workers' subjective mental workloads assessed by subjective rating of task difficulty (SRTD) were lighter than the students' at higher task loads. From the trend that there was no undue rise in either SRTD or percent increase in Oz beta 2 amplitude, it was judged that no task load could induce excessive mental workload on workers. PMID- 2572574 TI - Induction of glutamine synthetase and transient co-expression with carbamoylphosphate synthetase in hepatocytes transplanted into fat pads of syngeneic hosts. AB - Isolated rat hepatocytes were transplanted into the interscapular and both anterior lateral fat pads of hepatectomized syngeneic rats. At various time points following transplantation, the fat pads were removed, fixed and embedded in paraffin. Serial sections were stained for glutamine synthetase (GS) and carbamoylphosphate synthetase (CPS) using specific antisera and the PAP technique. The initially low fraction of GS+-heptatocytes remained low up to the fourth day, then increased strikingly up to almost 100% and declined gradually after the 14th day. In contrast, the number of CPS+-cells declined continuously to about 30% after 28 days. If the animals were exposed to CCl4 prior to the isolation of the hepatocytes in order to reduce the number of GS+-cells in the initial cell suspension similar results were obtained and no difference in the probability of the colony formation was noted between this and the normal hepatocyte suspensions indicating that the appearance of the GS+-phenotype was not due to a selective survival of these cells. Analysis of the staining intensity of the transplanted hepatocytes revealed the appearance of two populations of GS+-hepatocytes, one with a strong and one with a weak staining, during the course of formation of larger nodules, while only a single weakly stained population could be discerned with respect to the staining for CPS. These results demonstrate that all hepatocytes or at least their descendents can be induced to express GS by the environmental conditions of the fat pads, and that GS and CPS can be co-expressed with an apparently reciprocal relationship. PMID- 2572576 TI - Influence of infused beta-adrenergic agonists on porcine blood metabolites and catecholamines. AB - Anesthetized pigs were infused sequentially with increased concentrations of beta adrenergic agonists. At selected times during infusion, blood pressure, heart rate and plasma concentrations of free fatty acids (FFA), glycerol, glucose, lactate, norepinephrine, epinephrine and dopamine were measured. Azaperone, a drug used to calm the pigs before anesthesia, caused hypotension and bradycardia but did not affect plasma metabolites. Infusion of norepinephrine, epinephrine, isoproterenol or clenbuterol produced changes in plasma metabolites and plasma catecholamines. These changes during norepinephrine infusion were attributed to the infused agonist, whereas those during epinephrine infusion might have resulted to some extent from release of norepinephrine. Plasma isoproterenol was not quantified because it interfered with the assay of epinephrine and dopamine so that it was not possible to distinguish between infused isoproterenol and release of endogenous epinephrine and dopamine. Infusion of clenbuterol caused a small increase in plasma norepinephrine so that some of the increase in plasma FFA, glycerol and lactate during clenbuterol infusion may result from release of endogenous norepinephrine. PMID- 2572577 TI - Resistance to the Bacillus sphaericus toxin in cultured mosquito cells. AB - Increasing doses of Bacillus sphaericus toxin were used to select a toxin resistant cell line from the Culex quinquefasciatus line. This resistant cell line was proven to be C. quinquefasciatus in origin by isozyme analysis and karyotype. The resistant line bound fluorescent-labeled toxin as did the unselected susceptible line. A high level of resistance was quickly achieved, and this level was maintained after 4 mo. culture in the absence of toxin. PMID- 2572579 TI - Subdural empyema. PMID- 2572578 TI - Beta-adrenergic responsiveness in a human submandibular tumor cell line (A253). AB - Salivary epithelial functions are regulated by the autonomic nervous system. In this regard, we have been studying the morphology and neuroreceptor composition of A253, an immortal cell line isolated from a human submandibular carcinoma (Giard et al., JNCI, 51:1417-1421, 1973). Phase contrast and electron microscopic observations indicate that A253 cells are of epithelial origin. Physiologically, A253 cells possess beta-adrenergic, but not alpha-adrenergic or muscarinic cholinergic receptors. The beta-adrenergic receptors (BARs) are composed primarily of a single class of high affinity, beta 2-subtype receptors as judged by [3H]dihydroalprenolol antagonist binding studies. The BARs are functional inasmuch as isoproterenol stimulation increases both intracellular cAMP content and [3H]mannose incorporation into endogeneous glycoproteins. Differences in ultrastructure and neuroreceptor composition between A253 and other immortal salivary tumor cell lines are discussed. PMID- 2572580 TI - PapD, a periplasmic transport protein in P-pilus biogenesis. AB - The product of the papD gene of uropathogenic Escherichia coli is required for the biogenesis of digalactoside-binding P pili. Mutations within papD result in complete degradation of the major pilus subunit, PapA, and of the pilinlike proteins PapE and PapF and also cause partial breakdown of the PapG adhesin. The papD gene was sequenced, and the gene product was purified from the periplasm. The deduced amino acid sequence and the N-terminal sequence obtained from the purified protein revealed that PapD is a basic and hydrophilic peripheral protein. A periplasmic complex between PapD and PapE was purified from cells that overproduced and accumulated these proteins in the periplasm. Antibodies raised against this complex reacted with purified wild-type P pili but not with pili purified from a papE mutant. In contrast, anti-PapD serum did not react with purified pili or with the culture fluid of piliated cells. However, this serum was able to specifically precipitate the PapE protein from periplasmic extracts, confirming that PapD and PapE were associated as a complex. It is suggested that PapD functions in P-pilus biogenesis as a periplasmic transport protein. Probably PapD forms complexes with pilus subunits at the outer surface of the inner membrane and transports them in a stable configuration across the periplasmic space before delivering them to the site(s) of pilus polymerization. PMID- 2572581 TI - groE mutants of Escherichia coli are defective in umuDC-dependent UV mutagenesis. AB - Overexpression of the SOS-inducible umuDC operon of Escherichia coli results in the inability of these cells to grow at 30 degrees C. Mutations in several heat shock genes suppress this cold sensitivity. Suppression of umuD+C+-dependent cold sensitivity appears to occur by two different mechanisms. We show that mutations in lon and dnaK heat shock genes suppress cold sensitivity in a lexA-dependent manner. In contrast, mutations in groES, groEL, and rpoH heat shock genes suppress cold sensitivity regardless of the transcriptional regulation of the umuDC genes. We have also found that mutations in groES and groEL genes are defective in umuDC-dependent UV mutagenesis. This defect can be suppressed by increased expression of the umuDC operon. The mechanism by which groE mutations affect umuDC gene product function may be related to the stability of the UmuC protein, since the half-life of this protein is shortened because of mutations at the groE locus. PMID- 2572582 TI - Is the first enzyme of the shikimate pathway, 3-deoxy-D-arabino-heptulosonate-7 phosphate synthase (tyrosine sensitive), a copper metalloenzyme? AB - 3-Deoxy-D-arabino-heptulosonate-7-phosphate synthase (tyrosine sensitive) was purified from Escherichia coli carrying the plasmid pKB45. Enzyme of high specific catalytic activity (70 mu/mg) was obtained from cells grown only to the early log phase. The purified protein contained Cu(II) and showed an absorption band at 350 nm. Metal-free, catalytically inactive apoenzyme could be produced by dialysis against cyanide ion, and the holoenzyme could be reconstituted in terms of both catalytic activity and A350 by the binding of one Cu(II) ion per enzyme subunit. Zn(II) also reactivated the apoenzyme to about 50% of the level seen with Cu(II), although in this case no band appeared at 350 nm. In contrast to earlier reports that the enzyme contains substoichiometric levels of iron, insignificant amounts of iron were found in the isolated enzyme, and neither Fe(II) nor FE(III) regenerated either an absorption band at 350 nm or any catalytic activity from the apoenzyme. The evident preference of the enzyme as isolated for (Cu)II suggests that the synthase might naturally be a copper metalloenzyme. PMID- 2572584 TI - Nucleotide sequence of the glutamine synthetase gene (glnA) and its upstream region from Bacillus cereus. AB - We have determined the complete nucleotide sequence of a 2.4 kb chromosomal EcoT22I-NspV fragment, containing the Bacillus cereus glnA gene (structural gene of glutamine synthetase). The deduced amino acid sequence indicates that the glutamine synthetase subunit consists of 444 amino acid residues (50,063 Da). Comparisons are made with reported amino acid sequences of glutamine synthetases from other bacteria. Upstrem of glnA we found an open reading frame of 129 codons (ORF129) preceded by the consensus sequence for a typical promoter. Maxicell experiments showed two polypeptide bands, with molecular weights in good agreement with that of glutamine synthetase and that of ORF129, in addition to vector-coded protein. It is possible that the product of this open reading frame upstream of glnA has a regulatory role in glutamine synthetase expression. PMID- 2572583 TI - Purification and analysis of colonization factor antigen I, coli surface antigen 1, and coli surface antigen 3 fimbriae from enterotoxigenic Escherichia coli. AB - Enterotoxigenic Escherichia coli fimbriae are immunogenic and play a key role in intestinal colonization. Native colonization factor antigen I, coli surface antigen 1, and coli surface antigen 3 fimbriae were purified by a common method involving shearing, differential centrifugation, gel filtration, and density gradient ultracentrifugation. The compositions and N-terminal sequences were determined. Coli surface antigen 3 possesses two N-terminal isoforms, one of which matches the published DNA sequence, except for the previously proposed signal sequence cleavage point. PMID- 2572585 TI - Formation of malonyl coenzyme A in rat heart. Identification and purification of an isozyme of A carboxylase from rat heart. AB - Acetyl-CoA carboxylase is thought to be absent in the heart since the latter is highly catabolic and nonlipogenic. It has been suggested that the high level of malonyl-CoA that is found in the heart is derived from mitochondrial propionyl CoA carboxylase, which also uses acetyl-CoA. In the present study, acetyl-CoA carboxylase was identified and purified from homogenates of rat heart. The isolated enzyme had little activity in the absence of citrate (specific activity, less than 0.1 units/mg); however, citrate stimulated its activity (specific activity, 1.8 units/mg in the presence of 10 mM citrate). Avidin inhibited greater than 95% of activity, and addition of biotin reversed this inhibition. Further, malonyl-CoA (1 mM) and palmitoyl-CoA (100 microM) inhibited greater than 90% of carboxylase activity. Similar to acetyl-CoA carboxylase of lipogenic tissues, the heart enzyme could be activated greater than 6-fold by preincubation with liver (acetyl-CoA carboxylase)-phosphatase 2. The activation was accompanied by a decrease in the K0.5 for citrate to 0.68 mM. These observations suggest that the activity in preparations from heart is due to authentic acetyl-CoA carboxylase. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the preparation from heart showed the presence of one major protein band (Mr 280,000) and a minor band (Mr 265,000) while that from liver gave a major protein band (Mr 265,000). A Western blot probed with avidin-peroxidase suggested that both the 280- and 265-kDa species contained biotin. Antibodies to liver acetyl-CoA carboxylase, which inhibited greater than 95% of liver carboxylase activity, inhibited only 35% of heart enzyme activity. In an immunoblot (using antibodies to liver enzyme) the 265-kDa species, and not the major 280-kDa species, in the heart preparation was specifically stained. These observations suggest the presence of two isoenzymes of acetyl-CoA carboxylase that are immunologically distinct, the 265-kDa species being predominant in the liver and the 280-kDa species being predominant in the heart. PMID- 2572586 TI - Refined atomic model of glutamine synthetase at 3.5 A resolution. AB - An atomic model of 43,692 non-hydrogen atoms has been determined for the 12 subunit enzyme glutamine synthetase from Salmonella typhimurium, by methods of x ray diffraction including restrained least-squares atomic refinement against 65,223 unique reflections. At 3.5 A resolution the crystallographic R-factor (on 2 sigma data) is 25.8%. As reported earlier for the unrefined structure, the 12 subunits are arranged in two layers of six; at the interface of pairs of subunits within each layer, cylindrical active sites are formed by six anti-parallel beta strands contributed by one subunit and two strands by the neighboring subunit. This interpretation of the electron density map has now been supported by comparison with glutamine synthetase from Escherichia coli by the Fourier difference method. Each active site cylinder holds two Mn2+ ions, with each ion having as ligands three protein side chains and two water molecules (one water shared by both metals), as well as a histidyl side chain just beyond liganding distance. The protein ligands to Mn2+ 469 are Glu-131, Glu-212, and Glu-220; those to Mn2+ 470 are Glu-129, His-269, and Glu-357. The two layers of subunits are held together largely by the apolar COOH terminus, a helical thong, which inserts into a hydrophobic pocket formed by two neighboring subunits on the opposite ring. Also between layers, there is a hydrogen-bonded beta sheet interaction, as there is between subunits within a ring, but hydrophobic interactions account for most of the intersubunit stability. The central loop, which extends into the central aqueous channel, is subject to attack by at least five enzymes and is discussed as an enzyme "passive site." PMID- 2572587 TI - Cloning and analysis of the Neurospora crassa gene for cytochrome c heme lyase. AB - The cyt-2-1 mutant of Neurospora crassa is deficient in cytochromes aa3 and c and in cytochrome c heme lyase activity (Mitchell, M.B., Mitchell, H.K., and Tissieres, A. (1953) Proc. Natl. Acad. Sci. U.S.A. 39, 606-613; Nargang, F.E., Drygas, M.E., Kwong, P.L., Nicholson, D.W., and Neupert, W. (1988) J. Biol. Chem. 263, 9388-9394). By rescue of the slow growth character of the cyt-2-1 mutant, we have cloned the cyt-2+ gene from a N. crassa genomic library using sib selection. Analysis of the DNA sequence of the cyt-2+ gene revealed an open reading frame of 346 amino acids that has homology to the yeast cytochrome c heme lyase. The open reading frame is interrupted by two short introns. Codon usage and Northern hybridization analysis suggest that the cyt-2 gene is expressed at low levels. The cyt-2-1 mutant allele was cloned from a partial cyt-2-1 gene bank using the wild-type gene as a probe. Sequence analysis of the mutant gene revealed a 2-base (CT) deletion that alters the reading frame for 21 codons before generating an early stop codon in the protein-coding sequence. It was previously suggested that the cyt-2-1 mutation inactivates one of two regulatory circuits controlling the production of cytochrome aa3. The finding that the cyt-2-1 mutation affects the coding sequence for cytochrome c heme lyase provides a direct explanation for the deficiency of cytochrome c in the mutant and suggests that the lack of cytochrome aa3 is a regulatory response to the deficiency of cytochrome c. PMID- 2572588 TI - Modulation of the expression of a multidrug resistance gene (mdr-1/P glycoprotein) by differentiating agents. AB - Acquired resistance to multiple natural products in vitro is mediated by P glycoprotein (Pgp). Expression of this protein has been demonstrated in some normal tissues and in tumor samples obtained from both untreated and treated patients. In situ hybridizations with RNA probes have demonstrated higher levels of expression of mdr-1/Pgp in well-differentiated tumors and in well differentiated areas in tumors with mixed histologies. Expression of mdr-1/Pgp in human colon carcinoma cell lines was increased by the differentiating agents sodium butyrate, dimethyl sulfoxide, and dimethylformamide. In the SW-620 cell line addition of sodium butyrate resulted in a rapid induction of mdr-1/Pgp mRNA that was sustained for the duration of the exposure. The levels of P-glycoprotein were measured by immunoblotting and were also increased. Similar results were obtained in three other cell lines including the HCT-15 line. This induction occurred without alterations in nuclease sensitivity. Discontinuation of sodium butyrate was followed by a rapid fall in the levels of mRNA. The levels of P glycoprotein returned to normal with a half-life of about 24 h. In spite of a 20 25-fold increase in the level of mdr-1/Pgp mRNA and P-glycoprotein, the SW-620 cell line did not demonstrate increased resistance to doxorubicin and vinblastine or decreased accumulation of vinblastine. In contrast, in the HCT-15 cell line, a 5-fold increase of mdr-1/Pgp was accompanied by a comparable fall in vinblastine accumulation which was reversed by verapamil. In the SW-620 cell line, the induced protein could be photolabeled using [3H]azidopine. Expression of mdr 1/Pgp appears to correlate with the degree of differentiation. However, its induction is not always accompanied by expression of the multidrug-resistance phenotype. PMID- 2572589 TI - Increased activity, amount, and altered kinetic properties of IMP dehydrogenase from mycophenolic acid-resistant neuroblastoma cells. AB - Mouse wild-type neuroblastoma cells (NB cells) were stepwise selected for 10,000 fold increased resistance to mycophenolic acid (NB-Myco cells), an inhibitor of IMP dehydrogenase (IMP:NAD+ oxidoreductase, EC 1.1.1.205). IMP dehydrogenase activity was increased 25-fold, from 3.1 to 75 nmol/min.mg of protein; and a 56.7 kDa peptide was increased in abundance 200-500-fold in NB-Myco as compared to NB cells. Purification and sequence analysis confirmed that the abundant protein was IMP dehydrogenase. The stepwise selection, increased activity and protein abundance, and unstable phenotype are indirect evidence for a process of gene amplification. Kinetic findings consistent with an Ordered Bi Bi mechanism were indicative of IMP dehydrogenase having undergone mutation. The Michaelis constants were unchanged for IMP (14 and 13 microM) and increased 4-fold for NAD from 25 to 94 microM for NB and NB-Myco cells, respectively. The Ki for mycophenolic acid was increased 2400-fold from 1.4 nM to 3.4 microM for the enzyme from NB versus NB-Myco cells, and the Ki for XMP was increased 4-fold from 78 to 336 microM. Mycophenolic acid exhibited uncompetitive inhibition with IMP, consistent with the formation of a dead end E-XMP-inhibitor complex. The cellular GTP concentration was increased 2-fold in resistant cells and, upon removal of mycophenolic acid, further increased to 4.5-fold that of NB cells. PMID- 2572591 TI - Glycine to serine substitution in the triple helical domain of pro-alpha 1 (II) collagen results in a lethal perinatal form of short-limbed dwarfism. AB - Previous biochemical studies on cartilage tissue from a proband with Type II achondrogenesis-hypochondrogenesis (Godfrey, M., and Hollister, D. W. (1988) Am. J. Hum. Genet. 43, 904-913) indicated heterozygosity for a structural abnormality in the triple helical domain of pro-alpha 1 (II) collagen. Here we demonstrate that the mutation in the type II procollagen gene is a single base change that converts the codon for glycine (GGC) at amino acid 943 of the alpha 1 (II) chain to a codon for serine (AGC). The substitution disrupts the invariant Gly-X-Y structural motif necessary for perfect triple helix formation and leads to extensive overmodification, intracellular retention, and reduced secretion of type II collagen. These findings confirm the proposal that new dominant mutations in the type II procollagen gene may account for some cases of Type II achondrogenesis-hypochondrogenesis. Since recent studies (Lee, B., Vissing, H., Ramirez, F., Rogers, D., and Rimoin, D. (1989) Science 244, 978-980) have identified a dominantly inherited type II procollagen gene deletion in a non lethal form of skeletal dysplasia, namely spondyloepiphyseal dysplasia, the data more generally demonstrate that different type II procollagen gene mutations eventuate in a wide and diverse spectrum of clinical phenotypes. PMID- 2572590 TI - Hereditary alpha 2-plasmin inhibitor deficiency caused by a transport-deficient mutation (alpha 2-PI-Okinawa). Deletion of Glu137 by a trinucleotide deletion blocks intracellular transport. AB - alpha 2-Plasmin inhibitor is the most important physiological inhibitor of fibrinolysis; hence, its deficiency results in a severe hemorrhagic diathesis. We have cloned and characterized a mutant alpha 2-plasmin inhibitor gene from an individual homozygous for the deficiency. By sequencing all the coding exons and exon-intron boundaries of the gene, a trinucleotide deletion in exon VII that results in deletion of Glu137 was identified. We have designated this variant as alpha 2-plasmin inhibitor Okinawa. Using DNA samples amplified with the polymerase chain reaction, hybridization analysis by oligonucleotide probes confirmed the presence of this mutation in all the affected family members, including both parents. To elucidate the mechanism by which this mutation leads to the deficiency, a eukaryotic expression plasmid for alpha 2-plasmin inhibitor containing this mutation was constructed and transfected into COS-7 cells for transient expression analysis. Immunoprecipitation analysis and enzyme-linked immunosorbent assay revealed that the mutant alpha 2-plasmin inhibitor synthesized is mostly retained within the cells as an endoglycosidase H-sensitive form, and only a small portion of it is secreted into the medium as a neuraminidase-sensitive form. These results suggest that the Glu137 deletion leads to the alpha 2-plasmin inhibitor deficiency by causing a block in its intracellular transport from the endoplasmic reticulum to the Golgi complex. PMID- 2572593 TI - Endocytosis and Na+/solute cotransport in renal epithelial cells. AB - Endocytic uptake of [3H]sucrose and lucifer yellow, markers for fluid-phase endocytosis, was studied in cultures of the renal epithelial cell lines LLC-PK1 and OK. Endocytosis in LLC-PK1 cells was inhibited when the cells were grown in the presence of gentamicin (1 mg/ml) for 4 days or when the cells were treated with concanavalin A (1 mg/ml) for 5 h. These changes occurred without perturbation of intracellular Na+ and K+ content, indicating that the cells maintained normal ion gradients. The inhibition of endocytosis was accompanied by marked increases in the apparent Vmax for Na+-dependent cell uptake of solutes such as Pi and L-alanine. The apparent Km was unchanged. In contrast, treatment of OK cells with concanavalin A produced marked stimulation of endocytosis and inhibition of the Na+-dependent uptake of Pi and L-glutamate. These changes occurred in the absence of changes in intracellular Na+ and K+ content. Neither gentamicin nor concanavalin A had a direct effect on Na+/solute cotransport in these cell lines. The changes in Na+/Pi cotransport induced by concanavalin A in both LLC-PK1 and OK cells were blocked by keeping the cells at 4 degrees C during exposure to the lectin, suggesting that endocytosis may be part of the mechanism which mediates the changes in solute uptake. The reciprocal relationship between the changes in endocytosis and the changes in Na+/solute cotransport is consistent with the possibility that the number of Na+/solute cotransporters present in the plasma membrane may be altered by an increase or decrease in the rate of membrane internalization by endocytosis. The Vmax changes in Na+/solute cotransport provide indirect support for this conclusion. PMID- 2572592 TI - Sequential amplification of dihydrofolate reductase and multidrug resistance genes in Chinese hamster ovary cells selected for stepwise resistance to the lipid-soluble antifolate trimetrexate. AB - We describe the development of resistance to trimetrexate and piritrexim (BW 301U) by a stepwise selection protocol in Chinese hamster ovary cells. Selection in trimetrexate resulted in initial resistance as a result of dihydrofolate reductase gene amplification. Several trimetrexate-resistant variants that display 250-340-fold and 25-50-fold resistance to lipophilic and hydrophilic antifolates, respectively, were established. Increased antifolate resistance was associated with a prominent overexpression of dihydrofolate reductase as determined from the elevated folate reductase activity, cellular labeling with fluorescein-methotrexate, and steady-state mRNA levels as a result of a consistent dihydrofolate reductase gene amplification. However, upon subsequent incremental increases in trimetrexate, further resistance was also associated with amplification of the multidrug resistance gene. This resulted in overexpression of P-glycoprotein and a subsequent 20-50-fold collateral resistance to pleiotropic drugs such as adriamycin, actinomycin D, vinca alkaloids, etoposide, and colchicine. In contrast, initial resistance following selection with low piritrexim concentrations resulted from an unknown mechanism(s) not involving overproduction of either dihydrofolate reductase or P glycoprotein. This piritrexim resistance was shared with trimetrexate but not with methotrexate. Upon further selection with piritrexim, resistant variants emerge with amplified dihydrofolate reductase but not with multidrug resistance genes. These variants were subsequently resistant to both hydrophilic and lipophilic folate antagonists but retained sensitivity to pleiotropic drugs. The pattern of resistance with methotrexate, trimetrexate, and piritrexim shared a common mechanism, dihydrofolate reductase gene amplification, but differed regarding the additional amplification of the multidrug resistance gene in trimetrexate-resistant cells as well as the emergence of an additional unknown mechanism(s) of resistance to lipid-soluble antifolates upon initial selection in piritrexim. PMID- 2572594 TI - Site-specific substitution of glutamate for aspartate at position 59 of rat oncomodulin. AB - Replacement of the aspartate residue at position 59 of rat oncomodulin by glutamate by oligonucleotide-directed mutagenesis has afforded a protein which more closely resembles rat parvalbumin, at least judged by its interaction with the luminescent lanthanide ion Eu3+. The single-peak 7F0----5D0 spectrum observed at pH 5.0 with the fully bound wild-type protein is replaced by one which clearly shows two features at 5791 and 5796 A, arising from Eu3+ ions bound at the CD and EF sites, respectively. Furthermore, the pH dependence of the spectrum is substantially altered; the pKa observed for the CD domain, in which aspartate 59 residues, is shifted upward from pH 6.0 for the wild-type recombinant protein to pH 6.8 in the D59E mutant. Moreover, the maximum in the high-pH spectrum is shifted from 5781 to 5784 A. All three changes are indicative of a CD binding domain having increased parvalbumin-like character. Interestingly, however, the D59E substitution has only a modest effect on the Ca2+- and Mg2+-binding properties of the CD domain. For the wild-type protein, KCa = 7.8 x 10(-7) M and KMg = 3 x 10(-3) M. These affinities are more than an order of magnitude weaker than those seen for various parvalbumins and substantiate previous claims for calcium specificity made for the oncomodulin CD domain. Replacement of aspartate 59 by glutamate resulted in minor increases in affinity of the CD domain for Ca2+ (KCa = 5.5 x 10(-7) M) and Mg2+ (KMg = 1 x 10(-3) M). These findings strongly suggest that residues in oncomodulin besides aspartate 59 are important determinants of the observed calcium specificity of the CD calcium-binding domain. The consequences of the substitution at residue 59 appear to be confined to the CD domain. For the EF site in wild-type recombinant oncomodulin, KCa = 4.2 x 10(-8) M and KMg = 1.6 x 10(-4) M. The corresponding values for the D59E site specific variant are identical within experimental error (KCa = 4.2 x 10(-8) M and KMg = 1.8 x 10(-4) M). PMID- 2572595 TI - Cystine-rich type II antifreeze protein precursor is initiated from the third AUG codon of its mRNA. AB - The primary translation product encoded by sea raven antifreeze protein mRNA was labeled during cell-free synthesis with [3H]leucine. N-terminal sequencing of the immunoselected translation product showed that the third AUG in the mRNA is used as the initiating methionine codon. The antifreeze protein precursor is therefore 163 amino acids long. Amino acid analysis and sequencing of the deblocked N terminal peptide from the mature circulating form of the antifreeze indicated that glutamine at position 35 is the N-terminal residue. The most likely site of signal peptide cleavage is after alanine at position 17, suggesting that the sea raven antifreeze protein is produced as a preproprotein. Analysis of slot blots indicates that the gene for the antifreeze protein is present in 12-15 copies in the sea raven genome. A representative gene copy was sequenced. It is split into six exons spanning 2.2 kilobase pairs and, based on composite maps of genomic clones, is not accompanied by a second copy within at least 25 kilobase pairs of flanking DNA. The transcription start site was determined by primer extension. Ninety base pairs upstream from this point, beyond the CAAT and TATA boxes, is a putative cis-acting regulatory element in the form of a triplicated 21-base pair tandem repeat. PMID- 2572596 TI - Solubilization and immunopurification of a somatostatin receptor from the human gastric tumoral cell line HGT-1. AB - The human gastric tumoral cell line HGT-1 was previously shown to contain a membrane somatostatin receptor negatively coupled to adenylate cyclase through a pertussis toxin-sensitive inhibitory GTP-binding regulatory protein (Gi) (Reyl Desmars, F., Laboisse, C., and Lewin, M. J. M. (1986) Regul. Pept. 16, 207-215). In this study, we have solubilized this receptor in a free unoccupied form using Triton X-100 as detergent and [125I-Tyr11]somatostatin-14 to monitor specific binding. Furthermore, we have prepared a monoclonal antibody against a chromatographically enriched soluble receptor fraction and used this antibody (30F3) to immunopurify the receptor in conjunction with Sepharose-somatostatin-14 immunopurification and steric exclusion high pressure liquid chromatography (HPLC). The purified fraction showed 18,600-fold enrichment in terms of specific binding (i.e. from 0.6 +/- 0.05 to 11,300 +/- 830 pmol/mg of protein) and a single dissociation constant (kappa D) of 76 +/- 8 nM. On HPLC, it migrated as a single and symmetric 90-kDa peak. Moreover, after 125I-protein labeling, it gave a single 90-kDa band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis autoradiography. On the other hand, the 30F3 monoclonal antibody immunoblotted with a single 90-kDa band contained in the HGT-1 cell membrane. We therefore suggest that this antibody is specific to the HGT-1 membrane somatostatin receptor, that this receptor has a molecular mass of 90 kDa, and that we have obtained a homogeneous preparation of nondenatured receptor suitable for further cloning studies. PMID- 2572598 TI - Isolation and reconstitution of the chloride transporter of clathrin-coated vesicles. AB - Clathrin-coated vesicle acidification is mediated by an endomembrane proton translocating ATPase. This pump is electrogenic, and significant pH gradient formation requires the parallel movement of chloride through a chloride transporter in order to maintain net electroneutrality. We have solubilized, isolated and achieved 270-fold purification of this chloride transporter by means of selective detergent solubilization with cholate and polyoxyethelene 9-lauryl ether (C12E9), hydroxylapatite chromatography, and glycerol gradient centrifugation. Stabilization of the solubilized transporter requires 5 mM dithiothreitol. The partially purified transporter was co-reconstituted with the purified clathrin-coated vesicle proton translocating complex to yield preparations of proteoliposomes capable of valinomycin-independent proton pumping, as assessed by ATP-generated acridine orange quenching. In addition, the chloride transporter was independently reconstituted and was shown to catalyze diisothiocyano-disulfonic acid stilbene-sensitive 36Cl uptake. The anionic conductive selectivity of the reconstituted transporter (chloride = bromide greater than nitrate) exactly matched that of the transporter of native clathrin coated vesicles. These studies demonstrate that the chloride transporter of vacuolar acidification systems is structurally and functionally dissociable from co-existing proton pumps and allow for investigations of pump-transporter interactions in a resolved system. PMID- 2572597 TI - Effect of alpha-adrenergic blockers on calmodulin association with the nuclear matrix of rat liver cells during proliferative activation. AB - A transient peak of cytosolic calmodulin (CaM) was produced during the prereplicative phase of rat liver cell proliferation following partial hepatectomy. After accumulating in the cytosol, CaM apparently translocated into the nuclei, associating with the nuclear matrix. The administration of alpha 1 adrenergic blockers to hepatectomized rats prevented the association of CaM with the nuclear matrix without affecting the increase in the total nuclear CaM. The inhibitory effect of the alpha 1-antagonists was reversed by the simultaneous injection of the alpha-agonist noradrenaline. Since the activation of alpha 1 adrenergic receptors results in the release of Ca2+ from endoplasmic reticulum stores, the results suggest that the association of CaM with the nuclear matrix during proliferative activation is mediated by Ca2+ released from endoplasmic reticulum and show that the association with the matrix is independent of its intranuclear accumulation. PMID- 2572599 TI - Retinol-induced morphological changes of cultured bovine endothelial cells are accompanied by a marked increase in transglutaminase. AB - Retinol, a morphogen, has been shown to induce morphological changes in vascular endothelial cells, accompanied by an acute and specific accumulation of an 80-kDa protein; purification and characterization of this retinol-induced protein (RIP) have revealed that it is a transglutaminase. Endothelial cells from bovine carotid artery were cultured, treated with retinol, and examined for changes in morphology and protein profiles. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of extracts prepared from retinol-treated cells which had undergone a remarkable change in shape (from a cobblestone-like to a spindle like shape) indicated that the retinol-induced morphological change is accompanied by a marked increase of an 80-kDa protein. Similar changes were also induced by retinoic acid. The 80-kDa RIP was purified by anion exchange and hydroxyapatite column chromatography. Amino acid sequencing of tryptic fragments of the purified RIP revealed a high degree of homology between the sequence of bovine RIP and that of guinea pig liver transglutaminase, suggesting that RIP is a transglutaminase. This was confirmed by activity measurements; RIP exhibited transglutaminase activity, and an antiserum against RIP immunoprecipitated the activity. These results suggest that transglutaminase plays important roles in the maintenance of morphology and the control of endothelial cell functions. PMID- 2572600 TI - Tumor-related elevation of serum (alpha 1----3)-L-fucosyltransferase activity in gastric cancer. AB - (alpha 1----3)-L-Fucosyltransferase activity was measured in serum samples from 90 gastric cancer patients, 10 patients with benign diseases and 100 healthy controls. The enzyme activity was significantly elevated in the serum samples of patients with cancer compared to those from patients with benign diseases (P less than 0.01) and healthy controls (P less than 0.001). The elevation of the enzyme activity was found to correlate strongly with the clinical stage of disease. The sensitivity of (alpha 1----3)-L-fucosyltransferase was also demonstrated to be high in comparison with the tumor-associated antigens, such as carcinoembryonic antigen and sialylated Lewis X-i. Follow-up studies of (alpha 1----3)-L fucosyltransferase in 11 cancer patients with disease at different stages showed that the enzyme activity could be useful for monitoring the post-surgical course of the disease. These results suggest that (alpha 1----3)-L-fucosyltransferase activity has a clinically important potential as a tumor marker in gastric cancer. PMID- 2572602 TI - Mass spectral characterization of urinary pipamperone metabolites and high performance liquid chromatography assay for pipamperone plasma levels. PMID- 2572601 TI - Activation of the neu tyrosine kinase induces the fos/jun transcription factor complex, the glucose transporter and ornithine decarboxylase. AB - We have studied the ability of the neu tyrosine kinase to induce a signal for the activation of cell growth-regulated genes. Serum-starved NIH 3T3 cells expressing an epidermal growth factor receptor (EGF-R)/neu construct encoding a hybrid receptor protein were stimulated with EGF and the activation of the neu tyrosine kinase and stimulation of growth factor inducible genes were followed at the mRNA, protein, and activity levels, and compared to the corresponding responses in the neu proto-oncogene and oncogene expressing cells. Induction of the expression of jun mRNAs was an immediate early effect of EGF stimulation, followed by a marked increase in the biosynthesis of the fos/jun transcription factor complex and an increased transcription factor activity as measured by a recombinant transcription unit using chloramphenicol acetyltransferase assays. In distinction, elevated AP-1/PEA-1 activity in the absence of a significant increase in jun and fos expression was characteristic of the neu oncogene expressing cells. The glucose transporter mRNA increased at 2 h of EGF stimulation and was associated with enhanced glucose transport of the EGF-treated cells. An increase of ornithine decarboxylase (ODC) mRNA and activity followed these changes. In contrast, serum-starved, EGF-treated neu proto-oncogene- and oncogene-expressing cells showed constitutively low and high glucose transporter and ODC activities, respectively. These findings demonstrate that the chimeric EGF-R/neu receptor is capable of activating the expression of both immediate early genes and biochemical activities associated with cell growth stimulation. PMID- 2572604 TI - Use of a pilin gene probe to study molecular epidemiology of Pseudomonas aeruginosa. AB - Strains of Pseudomonas aeruginosa from patients with cystic fibrosis (CF) are unusual. The majority have a rough lipopolysaccharide (LPS) which renders them nontypeable by conventional typing systems based on a serological reaction with the O polysaccharide of smooth LPS. We developed a new typing scheme using a pilin gene probe as a marker for hybridization with endonuclease-digested genomic DNA from P. aeruginosa. Twenty-one different restriction fragment length polymorphism (RFLP) types were found among 249 isolates. RFLP type 7 was recovered only from patients with thermal burns (9 of 14 isolates) in both Vancouver, British Columbia, and Edmonton, Alberta, Canada. None of the other RFLP types showed a clear predilection for disease state or environmental niche. Multiple morphologically different isolates from individual patients with CF were studied; each isolate in 33 of 40 sputum samples had an identical RFLP type, despite considerable LPS serotype heterogeneity. Sequential isolates from 23 patients were studied; in 10 isolates there was a clear change in both the RFLP and the LPS serotype. We conclude that patients with CF usually harbor a single P. aeruginosa RFLP type in their sputa, but that one strain can replace another as the predominant colonizing type. PMID- 2572603 TI - Interactions of CD4+ and CD8+ human T lymphocytes from malaria-unprimed donors with Plasmodium falciparum schizont stage. AB - During Plasmodium falciparum malaria, a wide spectrum of parasite-encoded blood stage proteins is presented to the immune system of the host. To explore their multiple interactions with T cells from donors who have had no previous exposure to the parasite, whole schizont extract was used in vitro. Both CD4+ and CD8+ lymphocytes from all individuals tested were stimulated to proliferate. The responses were dependent on the presence of accessory cells and were only partially replaced by recombinant interleukin-1. Responses were inhibited by monoclonal antibodies to CD3, the alpha beta-chain T-cell receptor, or CD4 molecules but not to CD2. P. falciparum schizont extract-specific T-cell clones were generated and maintained by using sole stimulation by P. falciparum extract with autologous accessory cells or recombinant interleukin-2. Monoclonal antibodies to CD3 (or the alpha beta-chain T-cell receptor) blocked cloned T-cell responses to the schizont extract, and although the responses of the majority of the CD4+ or CD8+ T-cell clones were restricted by autologous accessory cells and inhibited by monoclonal antibodies to either CD4 or CD8, other clones responded to P. falciparum in the absence of accessory cells and were not regulated by the same monoclonal antibodies. The last category of clones consisted of autoreactive T cells. These data suggest that at the first contact with P. falciparum, requirements are met for significant T-cell stimulation. PMID- 2572605 TI - Neurotransmitter-lymphocyte interactions: dual receptor modulation of lymphocyte proliferation and cAMP production. AB - Stimulation of the beta-adrenergic receptor on lymphocytes can decrease the proliferative response of these cells to mitogens. We have found that simultaneous stimulation of T cells with the beta-adrenergic agonist isoproterenol and mitogens (phytohemagglutinin (PHA) and OKT3 monoclonal antibody) results in a 2- to 4-fold increase in cAMP production compared to cells exposed to isoproterenol alone. Mitogens alone have little effect on cAMP synthesis, but do activate the phosphatidylinositol (PI) cycle, suggesting that interactions may be occurring between the second messenger systems resulting in a cAMP synergy. Further experiments suggest that calcium may be involved in inducing the cAMP synergy observed in T cells. It is proposed that the synergy between beta-adrenergic and mitogenic stimulation of T cells for cAMP may be involved in the mechanism of catecholamine modulation of lymphocyte function. PMID- 2572606 TI - Bacterial cell surface hydrophobicity properties in the mediation of in vitro adhesion by the rabbit enteric pathogen Escherichia coli strain RDEC-1. AB - The role of hydrophobicity in the attachment of enteropathogens to gastrointestinal mucosa is controversial. In vitro binding of Escherichia coli RDEC-1 to rabbit intestine is dependent on the expression of pili. We examined in vitro adherence of piliated RDEC-1 after altering either the hydrophobicity of the organisms, the hydrophobicity of the substrate for attachment, or the surface tension of the suspending liquid. Hydrophobicity of RDEC-1 was determined using four complementary methods. In each assay piliated RDEC-1 demonstrated relatively more hydrophobic properties compared with both organisms grown to suppress pilus expression and a mutant that cannot express mannose-resistant pili. When piliated RDEC-1 were pretreated with tetramethyl urea to disrupt hydrophobic bonds surface hydrophobicity decreased. Concurrently, bacterial adherence to rabbit ileal microvillus membranes, mucus and mucin was reduced. Binding of piliated organisms to hydrophobic surfaces was significantly higher compared to both nonpiliated bacteria and the adherence of piliated RDEC-1 to relatively hydrophilic surfaces. Addition of propanol reduced the surface tension of the suspending liquid, and decreased adhesion of piliated RDEC-1 to polystyrene by 80%. Conversely, adherence of piliated organisms to a hydrophilic surface increased 12-fold after lowering the surface tension of the suspending liquid. We conclude that hydrophobic properties have a role in mediating in vitro adherence of this E. coli enteric pathogen. PMID- 2572608 TI - Effects of pentoxifylline on Ca2+-dependent transglutaminase in rat erythrocytes. AB - We have examined the effects of pentoxifylline, a drug used to improve peripheral blood flow in patients with chronic vascular disorders, on shear-induced Ca2+ entry and Ca2+-dependent protein crosslinking in the erythrocyte. Cells were washed in isosmotic bicarbonate buffer, pH 7.5, and brought to a 20% suspension. 5 ml samples were sheared by swirling them for seconds; 45Ca2+ uptake, activation of Ca2+-dependent transglutaminase, and products of the crosslinking reaction were measured. A 5 second shear promoted 45Ca2+ entry, activation of Ca2+ dependent transglutaminase and crosslinking of proteins in the membrane cytoskeletal fraction. Pentoxifylline, a drug that promotes erythrocyte deformability, diminished Ca2+ entry and inhibited activation of Ca2+-dependent transglutaminase, and had a prophylactic role on the effects of Ca2+ entry due to shear. Incubation of cells with 2.5 mM pentoxifylline before swirling minimized the effects of shear on 45Ca2+ entry, Ca2+-dependent transglutaminase and crosslinking of proteins. This study indicates that pentoxifylline promotes erythrocyte flexibility by minimizing shear-induced Ca2+ entry and Ca2+-dependent crosslinking. PMID- 2572607 TI - Most CD4+ T cells from human immunodeficiency virus-1 infected patients can undergo prolonged clonal expansion. AB - We have addressed the capacity of HIV-1 infection to alter the growth of primary CD4+ T cells, but at the clonal level. Single T cells were expanded in the presence of PHA, IL-2, and small numbers of accessory dendritic cells. We report two new findings. First, T cells from seropositive individuals, even those with AIDS and markedly reduced CD4+ counts, exhibit a normal cloning efficiency, and proliferative capacity. This result is in contrast to two prior reports of a low cloning efficiency in CD4+ T cells from HIV-1-infected patients. Second, when we added high doses of exogenous HIV-1 to T cell clones from control subjects, we observed infection but not cytotoxicity or loss of CD4+ cells, following addition of virus stocks at days 0, 3, and/or 7 of clonal growth. The same HIV-1 isolates markedly reduced CD4+ T cells in bulk mononuclear cultures. When tested at day 11, HIV-1 mRNA was expressed in some cells of exogenously infected clones by in situ hybridization; when tested at day 18, several clones could transactivate a TAT-sensitive cell line. These findings suggest that the loss of CD4+ T cells in infected individuals is not the inevitable result of the activation of latent infection, or spread of a productive infection, during clonal growth. PMID- 2572610 TI - The effect of triple drug therapy on renal function in patients with essential hypertension. AB - The effects of long-term triple drug therapy on renal function in patients with moderate to severe essential hypertension have not been evaluated systematically. We prospectively studied fifteen male patients with moderate to severe essential hypertension receiving triple drug therapy (metolazone, atenolol or betaxolol, and minoxidil) for 16 weeks. Supplemental potassium was prescribed in an attempt to maintain serum potassium above 3.5 mEq/liter. Systemic blood pressure was well controlled with this regimen. However, glomerular filtration rate (assessed by inulin clearance), effective renal plasma flow (assessed by paraaminohippurate clearance), and renal blood flow were reduced. Filtration fraction and renal vascular resistance were not significantly altered. Plasma renin activity remained stimulated throughout the protocol. Weight gain occurred, and serum potassium remained low. These results suggest that triple drug therapy employing a diuretic, beta-adrenergic antagonist, and a potent vasodilator is effective therapy for controlling moderate to severe systemic hypertension. However this antihypertensive regimen may be associated with a decrement in renal function. PMID- 2572609 TI - Regression of left ventricular hypertrophy in treated hypertensive patients with dilevalol and metoprolol--a double blind randomized study. AB - This study was designed to evaluate the effects of a new beta-adrenergic blocking agent with beta 2 agonist activity (dilevalol, an R-R' isomer of labetalol) on left ventricular hypertrophy regression as seen by M-mode echocardiography. The study design was a 2:1 double blind randomization of dilevalol versus metoprolol. There was an equal blood pressure reduction in the two groups (supine diastolic blood pressure fell from 101 +/- 4.5 mm Hg to 87 +/- 13.7 mm Hg, P less than .001 in the dilevalol group, and 101 +/- 4.3 mm Hg to 87 +/- 8.6 mm Hg, P less than .01, in the metoprolol group). At the end of 2 months, there was an overall 7.5% decrease in left ventricular mass index in the 16 dilevalol treated patients (this was due to a 4.4% decrease in posterior wall thickness, end diastolic dimension increased by only 1%). Of the seven patients with an increased left ventricular mass index, all demonstrated hypertrophy regression (mean 17.14%). In contrast, no significant change in left ventricular mass was seen in the metoprolol treated group. Echocardiographic left ventricular mass index and electrocardiographic evidence of left ventricular hypertrophy (using the Sokolow Lyons criteria and Romhilt-Estes point score) had poor correlation (r = .30 and r = .38, respectively). Resting ejection fraction increased by 5% and velocity of circumferential fiber shortening by 14% in the dilevalol treatment group (not significant).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572611 TI - The pharmacokinetics and pharmacodynamics of hydroxyzine in patients with primary biliary cirrhosis. AB - Hydroxyzine, a potent H1-receptor antagonist often used for relief of pruritus in patients with hepatic dysfunction, was studied in eight patients, mean age 53.4 +/- SD 11.2 years, with primary biliary cirrhosis. The patients ingested a single dose of hydroxyzine, 0.7 mg/kg (mean dose 43.9 +/- 6.6 mg). Before the dose, then hourly for 6 hours, every 2 hours from 6-12 hours, at 24 hours, and every 24 hours for 6 days, serum hydroxyzine and cetirizine were measured and an intradermal injection of 0.01 mL of a 0.1 mg/mL solution of histamine phosphate was performed. Wheals and flares were traced at 10 minutes and the areas were calculated. Mean peak hydroxyzine levels of 116.5 +/- 60.6 ng/mL occurred at 2.3 +/- 0.7 hours and mean peak cetirizine levels of 500.4 +/- 302.0 ng/mL occurred at 4.8 +/- 2.8 hours. The mean serum elimination half-life of hydroxyzine was 36.6 +/- 13.1 hours, and the mean serum elimination half-life of cetirizine was 25.0 +/- 8.2 hours. The mean hydroxyzine clearance rate was 8.65 +/- 7.46 mL/min/kg, and the mean volume of distribution was 22.7 +/- 13.3 L/kg. The mean wheal area was suppressed (P less than 0.01) from 1 to 120 hours, with maximal suppression from 2 to 48 hours. The mean flare area was suppressed from 1 to 144 hours, with maximal suppression from 3 to 24 hours (P less than 0.01). All patients became sleepy from 0.5 to 6 hours. Blurred vision, dizziness and dry mouth each occurred in two patients. Hydroxyzine elimination is impaired in patients with primary biliary cirrhosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572612 TI - Correlated light and electron microscopic study of dopaminergic neurons and their synaptic junctions with DARPP-32-containing cells in three-dimensional reaggregate tissue culture. AB - An antibody to tyrosine hydroxylase has been used in a correlated light and electron microscopic study to characterize dopaminergic neurons and synaptic junctions in three-dimensional reaggregate cell culture. Dissociated fetal mesencephalic cells containing dopamine neurons were coaggregated with dissociated fetal striatal cells in rotatory culture for 21 days. Sections of the coaggregates were stained by the peroxidase anti-peroxidase technique to reveal tyrosine hydroxylase-immunoreactive structures. Clusters of immunoreactive perikarya as well as dendrites and axons were observed. Immunolabeled perikarya were round or oval and approximately 20 microns in diameter. Boutons immunoreactive for tyrosine hydroxylase formed symmetric synapses, primarily with unlabeled dendritic shafts. Symmetric membrane specializations were also observed between tyrosine hydroxylase-positive boutons and unlabeled dendritic spines as well as with the perikaryon of an unlabeled medium-size neuron possessing a slightly indented nucleus. To characterize the neurochemical nature of the neurons postsynaptic to tyrosine hydroxylase-positive boutons in the reaggregates, an antibody against DARPP-32 (a dopamine and adenosine 3':5' monophosphate-regulated phosphoprotein) and an antibody against tyrosine hydroxylase were employed to visualize striatal dopaminoceptive neurons and dopaminergic structures, respectively, in the same section. Examination of reaggregate sections at the light microscopic level demonstrated that DARPP-32 immunoreactive cells were distributed into discrete clusters that were associated with patches of tyrosine hydroxylase-positive axonal varicosities. Ultrastructural analysis of tyrosine hydroxylase-positive boutons in such clusters revealed that dopaminergic axons synaptically contacted DARPP-32 immunoreactive neurons as well as unlabeled neuronal structures. PMID- 2572613 TI - Dopaminergic innervation of the basal ganglia in the squirrel monkey as revealed by tyrosine hydroxylase immunohistochemistry. AB - The organization of the dopaminergic mesostriatal fibers and their patterns of innervation of the basal ganglia in the squirrel monkey (Saimiri sciureus) were studied immunohistochemically with an antiserum raised against tyrosine hydroxylase (TH). Numerous fibers arose from midbrain TH-positive cell bodies of the substantia nigra pars compacta (group A9), the retrorubral area (group A8), and the lateral portion of the ventral tegmental area (group A10). These fibers accumulated dorsomedially to the rostral pole of the substantia nigra where they formed a massive bundle that coursed through the prerubral field and ascended along the laterodorsal aspect of the medial fore-brain bundle in the lateral hypothalamus. Some ventrally located fibers ran throughout the rostrocaudal extent of the lateral preopticohypothalamic area and could be followed up to the olfactory tubercle, whereas other fibers turned laterodorsally to invade the head of the caudate nucleus. At more dorsal levels in the lateral hypothalamus, many fiber fascicles detached themselves from the main bundle and swept laterally to reach the globus pallidus, the putamen, and the amygdala. Several TH-positive fibers coursed along the dorsal surface of the subthalamic nucleus, and some invaded the dorsomedial third of this structure. The remaining portion of the subthalamic nucleus contained relatively few TH-positive elements. In contrast, the globus pallidus received a dense dopaminergic innervation deriving mostly from two fascicles that coursed backward along the two major output pathways of the pallidum: the lenticular fasciculus caudodorsally and the ansa lenticularis rostroventrally. At the pallidal level, the labeled fibres merged within the medullary laminae and arborized profusely in the internal pallidal segment and less abundantly in the external pallidal segment. However, the caudoventral portion of the external pallidum displayed a dense field of TH-positive axonal varicosities. Other fibers ran through the dorsal two-thirds of the external pallidum en route to the putamen. The striatum contained a multitude of thin axonal varicosities among which a few long and varicosed fibers were scattered. These immunoreactive neuronal profiles were rather uniformly distributed along the rostrocaudal extent of the striatum but appeared slightly more numerous in the ventral striatum than in the dorsal striatum. The pattern of distribution of the TH-positive axonal varicosities in the dorsal striatum was markedly heterogeneous: it consisted of typical zones of poor TH immunoreactivity lying within a matrix of dense terminal labeling.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2572614 TI - Distribution and spatial geometry of dopamine interplexiform cells in the rat retina: I. Developing retina. AB - The morphology and distribution of dopaminergic interplexiform cells were analyzed in 9-day-old rat retinas processed as wholemounts for tyrosine hydroxylase immunohistochemistry. The mean number of dopaminergic interplexiform cells was estimated as about half of the total number of dopaminergic neurons in the immature retina, with a higher density in the temporal retina. Four interplexiform cells were individually analyzed and reconstructed with a computer system. Their arborizations could be divided into three different regions based on both their morphological features and their position within the retinal layers: (1) an internal arborization, spreading at the margin between the inner nuclear layer and the inner plexiform layer, composed of long, thick, somatofugal dendrites branching at acute angles, (2) an external arborization in the middle of the inner nuclear layer, formed by short, thin, varicose, recurved, axon-like processes branching at right angles, (3) and one or more scleral process(es), originating either from the cell body or from the internal arborization, running toward the outermost cell row of the INL, some of which reached the outer plexiform layer. Finally, analysis of the arborization network by computer simulations based on the 4 digitalized cells was compared with a nearest neighbour analysis of cell body distribution. It showed that cell bodies are almost randomly distributed--at least in the inferior retina--but that an adjustment of dendritic growth and orientation probably occurs to ensure a homogeneous coverage of the retina with a constant degree of overlap in the adult. This report represents the first three-dimensional computer reconstruction of chemically identified neurons in the retina. PMID- 2572615 TI - Ontogeny of catecholaminergic and cholinergic cell distributions in the cat's retina. AB - The development of catecholaminergic and cholinergic neurones in the cat's retina has been examined with antibodies against their respective rate-limiting enzymes, tyrosine hydroxylase (TH) and choline acetyl transferase (ChAT). ChAT immunoreactive (IR) cells were first detected at E (embryonic day) 56 with somata in the ganglion cell layer (GCL) or in the inner cytoblast layer (CBL). At P (postnatal day) 1, two faint bands of ChAT-IR fibres were evident in an inner and outer strata of the inner plexiform layer (IPL) and by P26, the bands were similar to those in the adult. TH immunoreactivity was first detected at E59 in either darkly labelled somata in the inner CBL with processes extending toward the IPL or in lightly labelled somata also located in CBL but with no processes. At P1, most TH-IR cells had prominently labelled dendrites and, by P8, most of the features of the adult cells were evident. Soma size gradients among TH-IR cells were first detected at P8, with cells in temporal retina being larger than those in nasal retina or at the area centralis. The smaller sizes of cells at the area centralis emerged after P26. The smaller sizes of ChAT-IR somata at the area centralis, by contrast, emerged between P8 and P26. The number of both TH-IR and ChAT-IR cells declined from the time they first appeared till adulthood. The decline was smaller among ChAT-IR cells (24%) than among TH-IR cells (68%). In distribution, the differential expansion of the retina appeared to be largely responsible for generating the final adult distribution of ChAT-IR cells. However, during late postnatal development (P26 to adulthood), the density of ChAT-IR cells in the periphery declined more than that of the ganglion cells, suggesting that some ChAT-IR cells may die in the periphery during this time. Prior to P26, the changes in the distribution of TH-IR cells were inconsistent with the pattern of retinal expansion. It is suggested that during this period, regional cell loss and cell addition may account for the changes in distribution of TH-IR cells. Later in development (P26 to adulthood), the changes in the density of TH-IR cells closely conformed to the differential expansion of the retina. PMID- 2572616 TI - The inhibitory actions of azelastine hydrochloride on the early and late bronchoconstrictor responses to inhaled allergen in atopic asthma. AB - We have examined the effect of azelastine hydrochloride, 8.8 mg, on the early and late responses to inhaled allergen in a group of 12 atopic subjects with asthma. On two separate days, 3 weeks apart, patients were administered either oral azelastine, 8.8 mg, or matched placebo. Four hours later they inhaled via nebulizer, a dose of allergen (grass pollen or Dermatophagoides pteronyssinus) that had previously been demonstrated to produce a 25% fall in FEV1. Plasma histamine concentrations and FEV1 levels were measured at intervals during the subsequent 8 hours. After placebo, allergen inhalation produced rapid bronchoconstriction in all subjects with a maximum mean fall in FEV1 at 30 minutes of 22.8 +/- 3.4% from the postsaline baseline value. Five subjects also developed a late bronchoconstriction response with a fall in FEV1 of greater than 15% from postsaline baseline value between 2 and 8 hours after challenge. Azelastine reduced the bronchoconstrictor response during the first 10 minutes and produced a maximum mean fall at 30 minutes of 21.2 +/- 4.4% from the postsaline baseline value. Azelastine had a marked inhibitory effect, reducing the maximum mean fall from 23.9 +/- 6.3% to 9.6 +/- 3.9% from the postsaline baseline value. Analysis of the area under the FEV1 response time-course curves revealed that azelastine reduced the early response (first 2 hours) by 32.5% (p less than 0.05) (all subjects) and reduced the late response (2 to 8 hours) by 70.2% (p less than 0.05) (n = 5). Azelastine had no significant inhibitory effect on the early increase in plasma histamine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572618 TI - The inhibitory effect of a selective alpha 2-adrenergic receptor antagonist on moderate- to severe-type asthma. AB - Midaglizole (DG-5128), a novel selective alpha 2-adrenergic receptor antagonist, was administered to 17 patients with moderate to severe asthma in a single-dose, double-blind, randomized, crossover study. All patients also continued their regular treatment regimens. FEV1 increased significantly (p less than 0.05), and respiratory resistance decreased significantly (p less than 0.05). Wheezing and dry rales on auscultation improved in patients receiving 200 mg of midaglizole, and their assessment of overall benefit was also significantly better (p less than 0.05). Blood pressure and heart rate were unaffected. However, plasma glucose levels fell slightly. This study demonstrated the bronchodilating effect of midaglizole in moderate to severe bronchial asthma. This selective alpha 2 adrenergic antagonist may prove to be a useful addition to current treatment regimens for asthma. PMID- 2572617 TI - Efficacy and safety of loratadine (10 mg once daily), terfenadine (60 mg twice daily), and placebo in the treatment of seasonal allergic rhinitis. AB - A total of 317 patients received loratadine, 10 mg once daily, terfenadine 60 mg twice daily, or placebo in a 14-day, double-blind, randomized study in seasonal allergic rhinitis. Four nasal and four nonnasal symptoms were evaluated. At the end point evaluation, mean total scores of combined nasal and nonnasal symptoms decreased from baseline (improved) 46%, 44%, and 35%, respectively, for loratadine, terfenadine, and placebo. The difference between loratadine and placebo treatment was significant (p = 0.03). Loratadine was particularly effective compared with placebo in relieving nasal discharge, sneezing, and itching/burning eyes. Therapeutic response to treatment was good or excellent in 66 (64%) of 103 loratadine-treated patients, 58 (56%) of 104 terfenadine-treated patients, and 48 (47%) of 102 placebo-treated patients. Adverse experiences reported during the study were usually mild or moderate and were not significantly different among the three treatment groups. Sedation (somnolence) was reported by 10 loratadine-treated patients, seven terfenadine-treated patients, and eight placebo-treated patients. Loratadine, 10 mg once daily, was comparable to terfenadine, 60 mg twice daily, and significantly superior to placebo in the symptomatic relief of seasonal allergic rhinitis. PMID- 2572620 TI - Neurophysiological regulation of the secretion of growth hormone. AB - The regulation of GH secretion involves the harmonious integration of factors acting at the hypothalamic and pituitary levels summating to produce the peripheral patterns of intermittent GH secretion observed in man and experimental animals under physiologic and experimental conditions. SRIH and GHRH act primarily at the pituitary to regulate the secretion of GH, while monaminergic, cholinergic and peptidergic neurotransmitters act centrally and/or at the pituitary level. In addition IGF-I, the anabolic effector of GH has inhibitory feedback effects at both the pituitary and hypothalamus. The control of GH secretion often differs between species and is dependent on the sex, nutritional state and physiologic condition being studied. In vivo and in vitro effects may differ as the complex, multilayered regulatory mechanisms of GH secretion in the intact subject override the direct effects of an individual controlling factor. PMID- 2572619 TI - The role of pharmacotherapy in anorexia nervosa and bulimia. AB - The purpose of this article is to review the basic pharmacology and the role of drugs that are used to treat anorexia nervosa and bulimia. The pharmacological treatment of eating disorders is based upon theoretical principles. The theoretical models include: (a) an illness secondary to other psychiatric disorders, (b) a disorder in the hypothalamic control of food intake, (c) a disorder of hypothalamic endocrine regulation, (d) a syndrome secondary to depressive illness, and (e) a disorder in the hypothalamic regulation of food intake. Theoretical models a, b, and c govern the choice of drug therapy for anorexia nervosa, and models d and e govern the choice of drug therapy for bulimia. Drugs used to treat anorexia nervosa and bulimia include tricyclic antidepressants and lithium carbonate. Chlorpromazine, metoclopramide, cyproheptadine, and clomiphene citrate have also been prescribed for the treatment of anorexia nervosa. Monoamine oxidase inhibitors are commonly prescribed to treat bulimia. Fenfluramine has the potential to be of therapeutic value in patients with bulimia. Although drug therapy plays a limited role in the treatment of eating disorders, drugs are commonly prescribed. Therefore, the nutritionist should be familiar with the basic pharmacology and the side effects related to drug therapy. PMID- 2572621 TI - Safety and efficacy of roxatidine acetate. Evidence from pharmacodynamic and clinical trials. AB - The effects of a new H2-receptor antagonist, roxatidine acetate, have been investigated in both clinical and pharmacodynamic trials in Europe and the United States. A series of four double-blind randomized studies are reviewed, reporting the effects of different dose regimens of roxatidine acetate compared with ranitidine and placebo in healthy volunteers using continuous intragastric pH monitoring. These pharmacodynamic studies clearly demonstrate that roxatidine acetate is an effective gastric antisecretory agent, which is up to twice as potent as ranitidine. The results of several clinical studies of roxatidine acetate in patients with gastric as well as duodenal ulcer conducted in Europe, Japan, and the United States are also reviewed. These studies show that roxatidine acetate is comparable to other potent H2-receptor antagonists in terms of cumulative healing rates, pain relief, and safety. Overall, the pharmacodynamic and clinical data indicate that the efficacy of roxatidine acetate 75 mg twice-daily (b.i.d.) does not differ significantly from ranitidine 150 mg b.i.d. Roxatidine acetate is equally effective in the treatment of peptic disease including gastric ulcer, duodenal ulcer, and reflux esophagitis. PMID- 2572622 TI - Pharmacokinetic characteristics of roxatidine. AB - This article reviews the published and unpublished results of pharmacokinetic studies with roxatidine acetate in healthy volunteers of different ethnic origins, patients with various degrees of renal impairment, patients on maintenance hemodialysis, lactating women, and elderly patients. In addition, it reports on the findings of interaction studies with food and other drugs. The pharmacokinetic characteristics of roxatidine were found to be nearly identical for different doses, formulations, and ethnic groups. The decrease in relative total clearance (oral clearance) in patients with renal impairment and in the elderly can be explained almost completely by their level of renal function. As expected from the pharmacokinetic characteristics in healthy volunteers, only a small amount of roxatidine is removed by hemodialysis. Dose reduction is recommended in patients with renal impairment, but dose supplementation after hemodialysis is not necessary. Only a negligible fraction of the dose administered is excreted with breast milk. No pharmacokinetic interactions were found with theophylline, warfarin, propranolol, diazepam, and desmethyldiazepam, antipyrine or antacids. Food did not interfere with the absorption or disposition of roxatidine. PMID- 2572623 TI - Gel diffusion determinations on feeding behaviour patterns of anophelines in Orissa State, India. AB - In 1981 Entomological Field Research Units were established in three districts of Orissa State, India, from which Anopheles bloodmeals were forwarded regularly for analysis to an Entomological Laboratory in Bhubaneswar. At the onset of 1982, the laboratory introduced a modified gel diffusion technique for determining the origin of mosquito bloodmeals and by the end of 1983, 22,300 smears had been processed and analyzed. The technique is relatively very simple, fast and inexpensive. The results are reproducible and the use of double controls (negative and positive) ensure a high degree of reliability. PMID- 2572624 TI - A device for separating mosquito larvae and pupae. PMID- 2572625 TI - Clotiazepam-induced acute hepatitis. AB - We report the case of a patient who developed acute hepatitis with extensive hepatocellular necrosis, 7 months after the onset of administration of clotiazepam, a thienodiazepine derivative. Clotiazepam withdrawal was followed by prompt recovery. The administration of several benzodiazepines, chemically related to clotiazepam, did not interfere with recovery and did not induce any relapse of hepatitis. This observation shows that clotiazepam can induce acute hepatitis and suggests that there is no cross hepatotoxicity between clotiazepam and several benzodiazepines. PMID- 2572627 TI - Development of erythromycin-resistance in Staphylococcus aureus as a consequence of high erythromycin consumption. AB - In a mixed ward for skin and venereology patients the erythromycin consumption was 6-8 fold higher than in other wards at the same hospital. Erythromycin resistance occurred in 15.5% of the infecting Staphylococcus aureus strains isolated from this ward, compared with 3.5% in other wards of the hospital and 2.3% in Denmark as a whole. The resistant strains belonged to different phage type patterns, but 18 out of 19 strains contained a 1.5 MDa plasmid and had an erythromycin-inducible resistance mechanism. Erythromycin resistance was frequent (approximately 10%) among Danish S. aureus strains in the years 1963 to 1969, mainly due to the spread of multiply-resistant strains of the 83A complex. During the last ten years, however, only 1-2% of the strains have been resistant, but a slow increase in resistance has taken place during the last three years among strains of all phage patterns including strains of group II, the 94, 96 complex and type 95. PMID- 2572626 TI - Demonstration of the cell cycle positions of taxol-induced "asters" and "bundles" by sequential measurements of tubulin immunofluorescence, DNA content, and autoradiographic labeling of taxol-sensitive and -resistant cells. AB - We used reliable and relatively inexpensive equipment to make sequential sets of measurements of antitubulin immunofluorescence, Feulgen staining, and autoradiography on the same cells. This was done to evaluate tubulin conformations, DNA content, and [3H]-thymidine incorporation in cell lines sensitive (HL60) and resistant (K562) to the novel anti-tubulin chemotherapeutic agent taxol. Numbers of cells with microtubule bundles have been found to correlate with sensitivity to taxol by clonogenic assay for several leukemic cell lines. We have found that cells with "asters" produced by taxol exposure are in mitosis and that cells with taxol-induced "bundles" are in G0/G1, S, and G2 phases. We further found that S-phase cells with microtubule bundles in both sensitive (HL60) and resistant (K562) cell lines were able to incorporate [3H] thymidine after 4-hr exposure to taxol. As microtubule bundles and asters occur in cells of the same cell cycle phases in both lines, we conclude that the greater frequency of cells with microtubule bundles reported for sensitive cells after taxol treatment cannot result from drug exclusion nor from different effects of the drug on cell microtubules in these two leukemic lines. PMID- 2572628 TI - Effect of mupirocin on nasal carriage of Staphylococcus aureus. AB - Mupirocin eliminates nasal carriage of Staphylococcus aureus among medical and surgical personnel for periods varying from several weeks up to one year. In persons recolonized after therapy densities of S. aureus populations in nares were much lower than in the same persons before therapy. PMID- 2572629 TI - Genetic heterogeneity in methicillin-resistant strains of Staphylococcus aureus revealed by esterase electrophoretic polymorphism. AB - 136 methicillin-resistant strains of Staphylococcus aureus recovered from hospitalized patients in 18 countries were characterized by electrophoretic mobilities of three types of esterases. These were defined by their ranges of activity toward five synthetic substrates and their resistance to di-isopropyl fluorophosphate. Fourteen zymotypes were distinguished. Two, designated as 6 and 14, were found in 53 and 50 strains, respectively. Genetic diversity coefficients were lower for strains from France and from other European countries (H = 0.47 and 0.53, respectively) than for strains from North America (H = 0.79). On the basis of electrophoretic polymorphism of esterases, our work provides evidence that methicillin-resistance is expressed in genetically different strains. Variations in esterase electrophoretic pattern within methicillin-resistant strains of S. aureus can make a significant contribution to the study of their epidemiology. PMID- 2572630 TI - Direct isolation of coagulase-negative staphylococci from neonatal blood samples. AB - Coagulase-negative staphylococci (CNS) were isolated by direct inoculation of anticoagulated whole blood onto agar from 14 (41.2%) of 34 episodes (30 neonates) of suspected bacterial infection, associated with isolation of CNS from the same blood sample by broth-dilution blood cultures. The equivalent of more than 1,000 cfu ml-1 were isolated from four samples (four neonates); the range of counts was 1-103 cfu 25 microliters-1. There was a statistically significant association between isolation of CNS by direct agar inoculation and a raised C-reactive protein level and/or a positive nitroblue tetrazolium test and with the use of an intravascular catheter when compared to those episodes where CNS were isolated from broth blood cultures only. This simple procedure provides an estimate of the number of CNS in the blood of a neonate with suspected bacteraemia and may help to corroborate a diagnosis of CNS bacteraemia. PMID- 2572631 TI - An in-use comparison of chlorhexidine gluconate 4% w/v, glycol-poly-siloxane plus methylcellulose and a liquid soap in a special care baby unit. AB - The effectiveness and harshness of three hand-wash agents, chlorhexidine gluconate 4%, glycol-poly-siloxane gel and a bland liquid soap were studied in use in a Special Care Baby Unit (SCBU). Each product was used for consecutive 2 week periods by the same groups of 8 health care workers among the 56 in the SCBU. The first group had normal skin, the second had 'sensitive skin' and the third group were known MRSA nasal carriers. Blood agar and methicillin mannitol salt agar were inoculated before and after hand washing and differences in colony counts calculated. Nursing activities before washing were graded from clean to dirty on an eight-point scale and correlated with before-wash colony counts. Product acceptability was assessed with a self-reporting questionnaire. Chlorhexidine was the most effective product in reducing skin microflora but users preferred to wash with the gel preparation. The sensitive-skin group achieved the best effects from washing, irrespective of the product used. Nasal carriers of MRSA recorded the lowest reduction levels of methicillin-resistant organisms but achieved the same reduction levels as the normal-skin group for other microflora. No relationship could be established between the pre-wash activity and the pre-wash colony count. The weekly MRSA colonization rate in neonates on the SCBU remained unaffected by any of the products tested. PMID- 2572632 TI - A comparison of four film-type dressings by their anti-microbial effect on the flora of the skin. AB - Two polyurethane dressings ('Tegaderm' and 'OpSite') were compared with their respective povidone iodine and chlorhexidine acetate-impregnated dressings ('Tegaderm Plus' and 'OpSite CH') for their effectiveness in reducing recolonization of skin after application of the dressings. After 7 days the average number of cfu on undamaged skin, covered with the four dressings, was significantly lower than the number of cfu on skin which had not been covered. The number of cfu on the skin covered with OpSite CH was significantly lower than with all other dressings tested. OpSite CH possesses most anti-microbial activity in relation to the flora of the skin. PMID- 2572633 TI - Do gowns prevent infection in neonatal intensive care units? AB - In a prospective study we found that gowns had no effect when used by medical and paramedical staff, in reducing infection in a tertiary care neonatal unit. We conclude that the use of gowns in this area is an ineffective and expensive method of preventing nosocomial infection. PMID- 2572634 TI - Assessment of the Axicare human milk pasteuriser (CM80). AB - The performance of the Axicare human milk pasteurizer was evaluated under different load conditions. The design criteria for pasteurisation specified that milk should be held at 63 degrees C for 30 min. The holding time at this temperature was insufficient if bottles of milk frozen at -20 degrees C were placed into the machine as directed, after the water-bath preheating phase. However, if the machine was loaded with frozen bottles before the whole cycle was commenced, adequate thawing and warming to holding temperature was achieved in sufficient time for satisfactory pasteurization. The pasteurizer gave consistent and adequate temperature and time conditions. The seals on three types of bottles tested tended to leak during the pasteurization cycle but no method of avoiding this problem was established. PMID- 2572635 TI - Hot-air hand driers. PMID- 2572636 TI - The effect of sodium alginate on the antibacterial activity of chlorhexidine, gentamicin and ciprofloxacin. PMID- 2572638 TI - Antibiotic resistance among gram-negative bacilli in 19 Greek hospitals. PMID- 2572637 TI - The potential hazard of using multiple-dose heparin and insulin vials in continuous ambulatory peritoneal dialysis. PMID- 2572639 TI - Control of an outbreak of nosocomial aspergillosis by laminar air-flow isolation. AB - In the course of construction of a new bone marrow transplantation (BMT) unit, six of nineteen children undergoing BMT in the adjacent ward area died of invasive pulmonary aspergillosis (IPA). Ward air samples confirmed that heavy fungal spore contamination had occurred. The introduction of laminar air flow (LAF) isolation terminated the outbreak and no subsequent cases of IPA have been documented in patients nursed entirely in LAF. LAF has not reduced the rates of other complications associated with BMT. PMID- 2572640 TI - Cytomegalovirus and legionella species as the cause of liver enzyme elevations in haemodialysis patients. AB - We conclude that reinfection or recurrent infection with CMV or legionella may be responsible for serum glutamic-pyruvic transaminase (SGPT) elevations in up to one fourth of haemodialysis patients. Since NANB infection or asymptomatic coinfection with these agents cannot be discounted with certainty, it seems prudent to maintain haemodialysis patients with abnormal liver-function tests on Body Fluid Precautions, even if they remain asymptomatic. PMID- 2572641 TI - An outbreak of multiply resistant Pseudomonas aeruginosa in a neonatal unit: plasmid pattern analysis. AB - An outbreak of infection due to multiply resistant Pseudomonas aeruginosa occurred from March to April 1986 in a neonatal unit. Affected neonates were receiving ventilation support and the mortality rate was high. Plasmid analysis and antibiograms indicated that the outbreak was due to a single strain. A survey of bacteria isolated from respirators, potable water and hands of personnel working in the unit failed to recover the outbreak strain. Lack of sterilization of respirators and overcrowding were considered to be the causes of the outbreak and reinforcement of the importance of aseptic techniques helped in its termination. PMID- 2572642 TI - Internalization of phosphatidylinositol-anchored lymphocyte proteins. I. Documentation and potential significance for T cell stimulation. AB - Several proteins that are anchored to the surface of T lymphocytes via a phosphatidylinositol (PI) moiety can initiate cell stimulation upon cross linking. Inasmuch as these proteins do not traverse the plasma membrane, it is not clear how they are capable of signaling across the membrane. Herein we report two distinct sets of experiments that examine the consequence of cross-linking PI anchored molecules on murine T cells. We first analyzed the fate of antibody cross-linked TAP (Ly-6A.2) and Thy-1 molecules on T-T hybrids. Using an assay to measure receptor-mediated endocytosis, an intracellular accumulation of 125I labeled anti-TAP and anti-Thy-1 mAb was documented that was specific and Ag dependent. The internalization of these molecules was confirmed by cytotoxicity assays using antibody-toxin conjugates, and electron microscopic studies. Although the PI-anchored proteins lack a cytoplasmic domain that is necessary for the internalization of many receptors, they nevertheless can be induced to enter the cell upon cross-linking. The rate of entry of cross-linked TAP and Thy-1 into cells was shown to be 10 and 2% per hour, respectively, which is considerably less than that observed for the transferrin receptor or TCR/CD3 complex. To assess whether the internalization of TAP and Thy-1 might be of importance in their ability to stimulate T cells, we attempted to cross-link these molecules under conditions where the mAb or its cross-linked complex can not enter the cell. We observed that anti-TAP and anti-Thy-1 mAb conjugated to a cell impermeant matrix fail to stimulate T cells. This loss of stimulatory activity was observed with multiple T-T hybridomas and mAb over a wide titration of antibody concentration and was independent of the mAb isotype. Results from experiments with anti-Ig cross-linking of the mAb-PI anchored protein complex suggested that the loss of T cell stimulation upon mAb immobilization is not simply due to an alteration in the degree of antibody cross-linking. These findings were generalized to three distinct PI-anchored proteins: TAP, Thy-1, and Ly6C on normal T cells. When the same cells were stimulated through the TCR/CD3 complex, only immobilized mAb are stimulatory. These results demonstrate a marked difference in the cross-linking requirements for stimulating T cells through PI anchored molecules in contrast to the transmembrane TCR complex. Furthermore, these findings raise the possibility that molecular internalization of Ab-PI anchored complexes may be necessary in signaling through these molecules. PMID- 2572643 TI - A T lymphoid cell line responds to a thymic stromal cell line by expression of Thy-1 and CD4. AB - We have cloned both T lymphoid and stromal lines from a single murine thymic tumor that was induced by a retrovirus carrying the v-myc oncogene (M-MuLV(myc]. The T lymphoid line, L4, was cloned by growth in agar. L4 cells were initially negative for Thy-1.2 and CD4 (although they contained rearranged TCR-beta genes), and they remained so if passaged in medium alone. However, cocultivation of these Thy-1.2- CD4- cells with the cloned stromal cell line, St3, resulted in sequential expression of Thy-1.2 and CD4 in subpopulations of cells. Thy-1.2+ CD4 and Thy-1.2+ CD4+ L4 subclones were obtained from the cocultures by subsequent cloning in agar. Derivation of these subclones from the starting Thy-1.2- CD4- clone was verified by Southern blot analyses specific for TCR-beta gene rearrangements and for M-MuLV(myc) proviral integration sites. Continuous cocultivation of Thy-1.2+ CD4+ L4 subclones with the St3 stromal cells was necessary for maintenance of CD4 on the cell surface. Furthermore, CD4 expression which was lost when CD4+ L4 cells were removed from the stroma could be reinduced if they were again cultured on St3 stroma. These cells may provide a model system for studying thymocyte-stromal cell interactions in induction and maintenance of expression of Thy-1 and CD4 molecules. PMID- 2572646 TI - IL-7 promotes thymocyte proliferation and maintains immunocompetent thymocytes bearing alpha beta or gamma delta T-cell receptors in vitro: synergism with IL-2. AB - IL-7 induced the proliferation of normal thymocytes and the effect was synergistically potentiated by a small dose of IL-2, which by itself hardly affected thymocyte proliferation. No synergism was observed between IL-7 and any one of the other lymphokines including IL-1, IL-3, and IL-4. The thymocyte culture stimulated with IL-7 and IL-2 consisted of single positive (CD4+CD8- and CD4-CD8+) and double negative (CD4-CD8-) populations, and double positive (CD4+CD8+) cells were completely deleted. Both single positive and double negative thymocytes expressed CD3, but only the former exhibited V beta 8 and V beta 6 in an expected proportion (approximately 30% in BALB/c mice) and the latter none at all. Immunoprecipitation of the cultured thymocytes by anti-TCR gamma antibody, on the other hand, revealed the presence of a TCR gamma chain. Taken together, these results indicated that the thymocyte cultured with IL-7 and IL-2 consisted of mature T cells bearing alpha beta or gamma delta TCR. Experiments using preselected thymocyte subpopulations indicated that double negative cells responded to both IL-7 and IL-2 with positive synergism when combined, while thymocytes enriched for single positive cells preferentially responded to IL-7 with little response to IL-2 and no detectable synergism. Double positive thymocytes showed no proliferation in response to IL-7 and IL-2. In contrast to single positive thymocytes, splenic T cells hardly responded to IL 7, although significant proliferation was induced in the presence of a low dose of IL-2. Thymocytes cultured with IL-7 and IL-2 showed little nonspecific cytotoxic activity, but responded to Con A or alloantigen, whereas those stimulated with a high dose of IL-2 alone exhibited potent cytotoxic activity. These results indicated that IL-7 was involved in the generation of immunocompetent T cells in the thymus in concert with IL-2. PMID- 2572645 TI - Down-modulation of MHC-I in a CD4+ T cell line, CEM-E5, after HIV-1 infection. AB - HIV-1 is capable of infecting many different cell types that express the CD4 molecule. In vivo and in vitro this infection is associated with profound immunologic defects. We have examined the effect of HIV-1 infection on the expression of MHC class I (MHC-I) molecules to explore the possibility that this important immune system molecule is perturbed after HIV-1 infection. Our data show that in vitro, HIV-1 infection of CD4+ PBL, and the CD4+ cell lines, CEM-E5, HT, and U937, results in decreased expression of MHC-I molecules on the cell surface. This down-modulation is transient, occurring 18 h after HIV-1 infection of CD4+ PBL and returning to normal expression by 24 h. In CEM-E5, MHC-I down modulation occurs over the course of days, reaching its greatest decrease (40%) about the time the cells are producing the most virus. Reversal of MHC-I expression to normal levels occurs as viral production decreases. Down-regulation during the time periods examined appear to be specific for MHC-I and does not occur with other cell-surface Ag nor is it caused by selection of a preexisting cell population with low MHC-I expression. Radioimmunoprecipitation of MHC-I protein from CEM-E5 indicated that the decrease of surface MHC-I is caused by decreased total protein secondary to a decrease in the level of mRNA for MHC-I. These decreased levels of MHC-I are biologically relevant because HIV-1 infected CEM-E5 cells are less susceptible to CTL lysis determined by the use of MHC-I cytolytic T cell clones and with the use of cold target-inhibition assay. PMID- 2572644 TI - Cellular basis of allograft rejection in vivo. V. Examination of the mechanisms responsible for the differing efficacy of monoclonal antibody to CD4+ T cell subsets in low- and high-responder rat strains. AB - MRC OX35, an anti-CD4 mAb, was used to treat high responder Wistar Furth (W/F) (RT1u) and low responder DA (RT1a) rats which had been grafted with directly vascularized hearts from PVG (RT1c) rats across a full MHC plus non-MHC incompatibility. Four doses of mAb at 7 mg/kg given in the first 2 wk postgrafting induced indefinite graft survival (greater than 150 days) in DA hosts, but only delayed rejection to 18 to 42 days in W/F as compared to rejection times of 6 to 8 days in untreated rats. The extension of MRC OX35 treatment to 6 wk in W/F rats induced indefinite graft survival in three of six rats. During treatment MRC OX35 therapy only partially depleted CD4+ cells, and all circulating CD4+ cells were coated with MRC OX35. The capacity of naive CD4+ and CD8+ cells from W/F and DA to be activated to PVG alloantigen was compared both in vitro in an MLC assay and in vivo by an adoptive transfer assay of their capacity to restore rejection of PVG heart grafts in irradiated syngeneic hosts. CD4+ cells from both W/F and DA proliferated in MLC and restored graft rejection. W/F CD8+ cells both proliferated in MLC and restored rejection, but DA CD8+ cells neither proliferated nor reconstituted rejection. Examination of lymphocytes from MRC OX35 treated hosts with long-surviving grafts showed that they were neither depleted of CD4+ T cells nor did they lack the capacity to proliferate to PVG Ag in MLC, this response being similar to that to third-party Ag or by naive lymphocytes. Compared to first-set rejection, PVG skin graft rejection was delayed 2 to 3 days in W/F and 10 to 12 days in DA rats with long-surviving grafts after MRC OX35 therapy, whereas they rejected third-party skin grafts in first-set tempo. These studies show that differences in graft survival in anti CD4 treated low and high responder strains may be due to the inherent capacity of CD8+ cells to be activated to effect rejection independent of CD4+ cells in W/F but not in DA. In those hosts that accept grafts, there is no evidence of clonal deletion, but there appears to be a form of unresponsiveness akin to that induced in adult rats by other immunosuppressive therapies that protects the graft from rejection. PMID- 2572647 TI - Human IgM monoclonal proteins that bind 3-fucosyllactosamine, asialo-GM1, and GM1. AB - Analysis of monoclonal human Ig that occur in association with lymphoproliferative diseases has provided valuable information about antibody structure and idiotypes. We analyzed 940 human sera that contained monoclonal IgM proteins for their ability to bind to four carbohydrate epitopes. Ten sera bound asialo-GM1, five of these sera also bound GM1, 10 bound to 3-fucosyllactosamine (3-FL), and one each bound to levan and galactan. Although the antibody activity in each serum was associated with a single L chain isotype, both kappa and lambda isotypes were represented among the proteins that bound to asialo-GM1 and to 3 FL. Some antibodies against asialo-GM1 were highly specific for this compound, whereas others cross-reacted with the structurally related gangliosides GM1 and GD1b. The antibodies to asialo-GM1 also varied considerably in their ability to lyse liposomes that contain asialo GM1. An association of IgM mAb against gangliosides with peripheral neuropathies has been reported recently, but only one of five patients whose antibodies reacted with GM1 ganglioside had a neuropathy. The antibodies that bound 3-FL exhibited narrower specificity, and less than 10% cross reactivity was noted with structurally related carbohydrates. The frequency of monoclonal proteins that bound 3-FL and asialo-GM1, approximately 1:100 sera for each specificity, was surprisingly high in view of the fact that both of these epitopes are expressed in human tissues. We suggest that these antibodies may be poly-specific and/or that the subset of B lymphocytes that synthesizes these anti-carbohydrate antibodies undergoes malignant transformation more frequently than other B lymphocytes. PMID- 2572648 TI - The physical separation of Lps and Ifa loci in BXH recombinant inbred mice. AB - Several reports in the literature suggest that many of the phenotypic defects of LPS-hyporesponsive C3H/HeJ mice may be attributed to decreased IFN production by their macrophages. The physical proximity on chromosome 4 of the gene which encodes sensitivity to LPS (Lps) and the structural gene cluster which encodes IFN-alpha (Ifa), suggests the possibility that the Lps gene, whose product is unknown, may actually be a part of the Ifa gene cluster. The C57BL/6J and C3H/HeJ mouse strains carry distinct alleles at both the LPs and the Ifa loci. In this study, we have analyzed these parental strains, as well as 12 recombinant inbred strains derived from these parental strains (e.g., BXH strains), for inheritance of these distinct alleles. The results show the segregation of these two loci: in 5 of 12 BXH RI strains, the IFN-alpha restriction fragment length polymorphism characteristic of one parental strain was discordant with the predicted LPS response phenotype. Therefore, we conclude that the Lps and the Ifa genes are physically distinct despite the apparent cause and effect relationship which is observed phenotypically. PMID- 2572649 TI - The extracellular acid phosphatase of the mosquito-parasitizing fungus Lagenidium giganteum. AB - The mosquito-parasitizing fungus Lagenidium giganteum secreted a soluble acid phosphatase and beta-D-glucosidase into the growth medium. The acid phosphatase was isolated and purified to single component, and some of its physicochemical properties were determined. The enzyme exhibited a pH optimum of 5.6 in phthalate buffer with p-nitrophenyl phosphate and was temperature-inactivated at 55 degrees C. Enzyme activity seems to be limited to phenyl-phosphate substrates. A molecular weight of 42,800 was found and the amino acid content was also determined. A Km for p-nitrophenyl phosphate of 1.6 x 10(-7) M was found. The possible involvement of the enzyme in the infective process was discussed. PMID- 2572650 TI - Response to pneumococcal vaccine among asymptomatic heterosexual partners of persons with AIDS and intravenous drug users infected with human immunodeficiency virus. AB - Antibody responses to pneumococcal vaccine were studied in asymptomatic heterosexual partners of persons with AIDS and intravenous drug users seropositive for human immunodeficiency virus (HIV). Serum antibodies to 12 pneumococcal capsular antigens were measured by radioimmunoassay. Eleven intravenous drug users seropositive for HIV, 13 seronegative intravenous drug users, and 10 each seropositive and seronegative sexual partners received 23 valent pneumococcal vaccine. Additional unvaccinated matched seropositives served as controls. Antibody responses were significantly lower among subjects seropositive for HIV (P less than .05). Fourteen (88%) of 16 seropositive subjects with baseline type-specific antibodies to one or more pneumococcal antigens less than 300 ng of antibody nitrogen/ml (ngAbN/ml) demonstrated a rise in one or more of these antibodies to greater than 400 ngAbN/ml. No clinical deterioration or decrease in T4 cells attributable to vaccination was found. Although antibody responses to pneumococcal vaccine among HIV-infected subjects were impaired, most with antibody levels less than 300 ngAbN/ml developed titers of one or more type-specific antibodies to levels greater than 400 ngAbN/ml without notable adverse effects. PMID- 2572651 TI - Serum IgG, IgA, and IgM responses to pertussis toxin, filamentous hemagglutinin, and agglutinogens 2 and 3 after infection with Bordetella pertussis and immunization with whole-cell pertussis vaccine. AB - Class antibody responses to pertussis toxin (PT), filamentous hemagglutinin (FHA), and agglutinogens 2 and 3 (Agg 2/3) were studied in 114 patients with pertussis, 49 family contacts of patients with pertussis, and 139 infants attending vaccination clinics. IgG responses to FHA and PT were similar in patients with pertussis and in infants immunized three times with whole-cell pertussis vaccine. IgA responses to FHA and PT were greater in patients with pertussis than in vaccines, and a serum log10 titer of IgA to PT greater than 1.9 was a useful discriminant of Bordetella pertussis infection. When compared with patients with pertussis, vaccinees had a greater IgG response to Agg 2/3 and an equivalent IgA response to Agg 2/3. Serologic responses to FHA, PT, and Agg 2/3 were common in family contacts but were smaller than the responses in patients with pertussis. Initial titers of IgG to FHA and IgA to FHA were significantly higher (P less than .005 and P less than .05, respectively) in family contacts aged 3-10 y than in patients with pertussis in an identical age group. PMID- 2572652 TI - Influence of antihistamine pretreatment on vancomycin-induced red-man syndrome. AB - Twelve adult male volunteers participated in a double-blind study to determine the effect of pretreatment with histamine and/or histamine2 receptor antagonists on the incidence and severity of vancomycin-induced "red-man syndrome." Subjects received hydroxyzine, 50 mg, ranitidine, 300 mg, hydroxyzine plus ranitidine, or placebo at weekly intervals 2 h before a l-h infusion of vancomycin, 1,000 mg. The extent of erythema was evaluated by burn chart and pruritus was assessed by each subject using a rank scale; symptoms were thus classified a priori as mild, moderate, or severe and their summation determined a global severity score for red-man syndrome. There was no significant intrasubject variation between area under the concentration-time curves for histamine and vancomycin (P greater than .05). Pretreatment with hydroxyzine alone provided significant protection against vancomycin-induced erythema and pruritus (P less than .05) whereas the effect of ranitidine did not differ significantly from placebo. The combination of hydroxyzine and ranitidine offered no advantage over hydroxyzine alone. Similar results were observed for global severity scores. Thus in normal volunteers prior administration of a histamine antagonist (hydroxyzine) provided significant protection against vancomycin-induced red-man syndrome compared to that afforded by placebo. There was no significant benefit from the histamine antagonist alone or by its addition to a histamine antagonist. PMID- 2572654 TI - [The clinical evaluation of sialyl Lewis Xi antigen in patients with gynecologic tumors]. AB - In order to estimate the clinical significance of sialyl Lewis Xi (SLXi) antigen, the antigen was measured with an "FH-6" Otsuka Kit in sera from patients with various gynecologic tumors and healthy women. The antigen in ovarian cyst fluids was also determined. Furthermore, serum SLXi antigen levels were serially followed up in the patients with elevated serum SLXi levels to evaluate the correlation between serum SLXi levels and the response to treatment. Results obtained were as follows. 1) Among the patients with uterine myoma, uterine malignancies and benign ovarian tumors, the incidence of elevated serum SLXi antigen levels was very low. 2) Among the patients with ovarian malignancies, serum SLXi antigen levels was significantly increased in the following order: clinical stage I (38%), stage II (50%) and stage III (65%). 3) The high SLXi value was observed in the cyst fluid from all ovarian mucinous adenomas and all ovarian cancers. In addition SLXi antigen level was significantly higher in the cyst fluid of mucinous adenocarcinomas than that of serous cystadenocarcinomas. 4) Serum SLXi values were correlated with the effect of treatment. Interestingly, the elevation of serum SLXi levels preceded the clinical detection of recurrence by 3 month in a case. Thus SLXi antigen appears to be a useful marker for monitoring of ovarian malignancies, especially of mucinous adenocarcinomas. PMID- 2572653 TI - Hepatitis A virus infection in a chimpanzee: duration of viremia and detection of virus in saliva and throat swabs. AB - The pathogenesis of hepatitis A virus was studied by orally inoculating a chimpanzee with wild-type virus. The virus was initially detected in the animal's serum (day 14 after inoculation); then viral antigen was detected in the tonsils (day 16), virus in the saliva and throat swabs (day 18), and viral antigen in the liver (day 21). The animal was viremic for 2 w. While this study involved a single animal, the presence of hepatitis A virus in saliva and throat swabs suggests a possible oropharyngeal site for viral replication. PMID- 2572655 TI - [Clinical evaluation of serum CA130 in patients with lung cancer]. AB - CA130 is a glycoprotein which is recognized by monoclonal antibodies-(130-22 and 145-9) produced by immunization with human lung adenocarcinoma cell line (PC-9). CA130 is considered to be a new tumor marker, different from CA125, since it has a separate antigenic determinant. We used the D-7111 kit (Daiichi Radioisotope Laboratories, Ltd.) to measure the serum level of CA130 in 290 patients with lung cancer, 171 patients with noncancerous lung disease (N-CLD) and 93 healthy adults. In addition, CEA, CA19-9, CA125 and sialyl SSEA-1 antigen (SLX) were also measured for the same serum samples when possible. The cutoff level for CA130 was 35 U/ml. The overall positive rates for CA130 were 32% in the lung cancer patients, 23% in the N-CLD patients and 0% in the healthy adults. The positive rate in the lung cancer patients was significantly higher than in the N-CLD patients (p less than 0.05). As a function of the histological type of lung cancer, the positive rates for CA130 were 44% in 120 patients with adenocarcinoma, 17% in 115 patients with squamous cell carcinoma, 33% in 36 patients with small cell carcinoma, 42% in 12 patients with large cell carcinoma and 29% in 7 patients with miscellaneous cell type. The positive rate in the patients with adenocarcinoma was significantly higher than in the patients with squamous cell carcinoma (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572656 TI - [Basic and clinical investigations on pituitary gland]. PMID- 2572657 TI - [A case of autonomic hyperreflexia successfully treated with alpha-adrenergic blocking agent]. PMID- 2572658 TI - Quantitation of the host cell infiltration kinetics of the nonimmunogenic colon 26 tumor by multiparameter flow cytometry. AB - In this study, we examined the kinetics of host cell infiltration into the nonimmunogenic Colon 26 tumor. We found that 1 x 10(4) cells were required to produce tumors in 100% of mice. The vivo doubling time was 42.5 h, and a barely palpable tumor contained 8 x 10(6) cells. No evidence of concomitant immunity was found. The number of host cells infiltrating the in vivo tumors increased at the same rate as the number of tumor cells, but averaged only 22% of total cells. Cycling T lymphocytes were present in the host cell infiltrate of this tumor. In addition, approximately 50% of in vivo Colon 26 cells were Thy-1.2 positive. The observed characteristic of low immunogenicity makes it a useful murine model for studying human malignant tumors. PMID- 2572659 TI - Heterogeneity of macrophages in the rat evidenced by variability in determinants: two new anti-rat macrophage antibodies against a heterodimer of 160 and 95 kd (CD11/CD18). AB - A set of three monoclonal antibodies (MoAbs), ED1, ED2, and ED3, has been shown to recognize in situ different subsets of macrophages in the rat. This macrophage diversity can be correlated with differences in stage of differentiation of cells belonging to one lineage. The present study quantifies this antigen distribution in the macrophage fractions of several lymphoid organs provided by Percoll centrifugation. Four new MoAbs (ED4, ED7, ED8, and ED9) raised against macrophages are included in this study. The tissue distribution of each of the four new MoAbs is determined by immuno- and enzyme-histochemistry on cryostat sections. The MoAbs recognize distinct subpopulations of macrophages. The new MoAbs ED4, ED7, ED8, and ED9 recognize granulocytes and other unrelated cell types, as well as cells of the mononuclear phagocyte system. ED7 and ED8 recognize a surface heterodimer of Mr 160,000 and 95,000. PMID- 2572660 TI - Antihypertensive therapy with guanfacine induces elevated plasma growth hormone levels in diabetic patients. AB - Guanfacine, a central alpha-adrenoreceptor agonist, may increase growth hormone (GH) secretion. We have investigated the effect of guanfacine upon plasma GH levels in 16 hypertensive non-insulin-dependent diabetic (NIDD) patients. Guanfacine (1 to 2 mg/day/os) was administered for six months, and GH measured in basal plasma samples before and after treatment with this drug. Our results show an increase in GH plasma levels after guanfacine administration (2.88 +/- 2.05 ng/ml (X +/-SD) before, and 4.37 +/- 1.80 ng/ml (X +/- SD) after treatment). Since it is believed that GH levels plays a role in the course of diabetic retinopathy, caution should be taken with guanfacine antihypertensive treatment in patients affected with diabetes mellitus. PMID- 2572661 TI - Adrenergic urticaria: a first report from Thailand. AB - A 22-year-old, female patient came to see the allergists because of a 10-year history of chronic urticaria. Widespread pruritic, urticarial papules developed at times of stress and exercise, each papule being surrounded by a striking blanched vasoconstricted halo. The halo hives could be replicated with an intradermal injection of adrenaline. This is the first report of adrenergic urticaria from Thailand. PMID- 2572662 TI - Effect of acetylcholine and 5-hydroxytryptamine on the secretion of corticotrophin-releasing factor-41 and arginine vasopressin from the rat hypothalamus in vitro. AB - Previous studies using the isolated rat hypothalamus in vitro have shown that both acetylcholine and 5-hydroxytryptamine (5-HT) stimulate the secretion of bioactive corticotrophin-releasing factor (CRF). However, the CRF complex consists of a number of bioactive substances, and in this study we examined the effect of acetylcholine and 5-HT on the release of immunoreactive (ir)-CRF-41 and ir-arginine vasopressin (AVP) in vitro. Acetylcholine caused a dose-dependent (10 pmol-10 nmol/l) release of both neuropeptides, and the effect was partially antagonized by both atropine and hexamethonium. Nicotine (0.1-10 mumol/l) also stimulated the release of both peptides, whereas bethanacol had no effect on AVP release, but had a variable action on CRF-41 release. 5-HT caused a dose dependent (10 pmol-1 nmol/l) stimulation of CRF-41 release without any effect on AVP release, and this effect was antagonized by cyproheptadine, suggesting the participation of specific 5-HT receptors. PMID- 2572663 TI - Thyroid hormone affects the development of Sertoli cell function in the rat. AB - The relationship between thyroid activity and Sertoli cell function has been investigated in prepubertal rats. Male 28-day-old Wistar rats were used to prepare Sertoli cells by sequential enzyme digestion of the testes. Hypothyroidism, induced by oral administration of methimazole from the day of birth, was characterized by a severe retardation of body and testis growth and a net inhibition of the increase in Sertoli cell gamma-glutamyl transpeptidase (GGT) activity as well as in androgen-binding protein (ABP) and lactate production, which normally occur during postnatal development of Sertoli cells. The functional parameters of Sertoli cells from hypothyroid 28-day-old rats approximated to those of cells from euthyroid 15-day-old animals. These results are consistent with the impairment of protein synthesis in Sertoli cells from hypothyroid rats compared with controls. Body and testis growth were improved and Sertoli cell functions were restored with 3,3',5-tri-iodothyronine (T3) replacement therapy. An excess of T3 in the serum, induced by daily i.p. injections of T3 (100 micrograms/kg body wt) during the last week before the rats were killed, failed to induce changes in body and testis growth or in the activity of GGT and lactate dehydrogenase of Sertoli cells. Cells from hyperthyroid rats exhibited a specific decrease in ABP production. These results indicate that thyroid hormone is necessary for the postnatal maturation of Sertoli cell function and suggest a regulatory role of the hormone on gametogenic development in the prepubertal rat. PMID- 2572664 TI - Effects of growth hormone-releasing hormone on the secretion of islet hormones and on glucose homeostasis in lean and genetically obese-diabetic (ob/ob) mice and normal rats. AB - The effect of synthetic human growth hormone-releasing hormone(1-40) (hGHRH-40) on the function of the endocrine pancreas and on glucose homeostasis in lean and genetically obese-diabetic (ob/ob) mice and normal rats has been examined. The addition of 1 mumol hGHRH-40/1 to incubated islets from normal lean mice increased insulin release by 90 and 37% at 5.6 and 16.7 mmol glucose/l respectively. Lower concentrations of hGHRH-40 did not affect insulin release. hGHRH-40 (1 mumol/l) increased pancreatic polypeptide release by 50% at 5.6 mmol glucose/l. A range of concentrations of hGHRH-40 (1 nmol/l-1 mumol/l) reduced glucagon release by 42-73% at 5.6 mmol glucose/l, and by 38-70% at 16.7 mmol glucose/l. Somatostatin release was increased (eightfold) by 1 mumol hGHRH-40/1 at 5.6 mmol glucose/1, but at 1 nmol hGHRH-40/1 somatostatin release was reduced (by greater than 50%). At 16.7 mmol glucose/litre 0.01-1 mumol hGHRH-40/1 increased somatostatin release (three- to fourfold), but 1 nmol hGHRH-40/1 produced a reduction of 50%. In vivo, administration of hGHRH-40 (50 micrograms/kg body weight i.p.) to fasted lean and ob/ob mice did not alter basal plasma concentrations of glucose and insulin, or the glucose and insulin responses to a concomitant i.p. glucose challenge. Intravenous injection of hGHRH 40 (20 micrograms/kg body weight) to anaesthetized rats increased plasma concentrations of insulin in the hepatic portal vein. A lower dose of hGHRH-40 (0.2 micrograms/kg) was ineffective, and neither dose of hGHRH-40 altered plasma glucose.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572665 TI - Relationship between growth hormone-releasing hormone and somatostatin in the rat: effects of age and sex on content and in-vitro release from hypothalamic explants. AB - Secretion of GH in the rat has been shown to be dependent upon age and sex. Using rat hypothalamic explants in vitro, we have studied the release and hypothalamic content of GH-releasing hormone (GHRH) and somatostatin in male and female Wistar rats at four different ages (10, 30 and 75 days, and 14 months). Basal release of GHRH was not significantly different between male and female rats, but at all ages males released more GHRH in response to stimulation by both 28 and 56 mmol potassium/l than female rats (P less than 0.05). Neither basal nor potassium stimulated release of GHRH altered with age. In contrast, both basal and potassium-stimulated secretion of somatostatin increased significantly (P less than 0.01) with age, but was the same in the two sexes. Hypothalamic GHRH content, as assessed by the extractable tissue content following incubation, was significantly (P less than 0.01) lower in 10-day-old rats compared with older rats, but remained constant after 30 days of age. Somatostatin content, in contrast, increased progressively with age (P less than 0.01). The hypothalamic content of the two peptides was the same in both sexes. In conclusion, our findings demonstrate that male rats release more GHRH in vitro than female rats, possibly reflecting the increased pulse amplitude of GH seen in males in vivo; the progressive fall in secretion of GH previously reported during ageing appears to parallel the progressive increase in somatostatin release and content seen in our in-vitro system. PMID- 2572667 TI - Relationships between serum triglycerides and gamma-glutamyltransferase among shift and day workers. AB - Increased levels of serum triglycerides in shift workers have been reported in two population-based studies. In order to examine this relationship and to take some common confounding factors into account this cross-sectional study of 562 shift and day workers was carried out. The shift workers had significantly higher levels of serum triglycerides. gamma-glutamyltransferase and total cholesterol, and Body Mass Index did not differ between the groups. The cause of increased serum concentrations of triglycerides is unclear. The present study provides no evidence to support the assumption that alcohol drinking or obesity accounts for increased levels of serum triglycerides in shift workers. PMID- 2572666 TI - Effect of xamoterol on exercise capacity and left ventricular function in angina pectoris and in dilated cardiomyopathy. AB - The present study evaluates the effect of the beta 1 partial agonist xamoterol in patients with coronary artery disease compared to the effect in patients with dilated cardiomyopathy. Nineteen patients with angina pectoris and nine patients with dilated cardiomyopathy received xamoterol 200 mg b.i.d. for 4 weeks in a placebo-controlled double-blind study. The effect was evaluated with a standardized exercise test and radionuclide angiocardiography with determination of global and regional ejection fraction. At rest xamoterol caused no significant changes among the angina patients while global and regional ejection fraction increased among the cardiomyopathy patients. During exercise, when the drug exerts a beta 1-antagonist effect, xamoterol increased the exercise performance among the angina pectoris patients. Fourteen out of 19 patients with angina pectoris were unable to increase their ejection fraction from rest to work due to coronary ischaemia. These 14 patients improved their global ejection fraction during work after xamoterol treatment. PMID- 2572669 TI - Evidence for mouse Th1- and Th2-like helper T cells in vivo. Selective reduction of Th1-like cells after total lymphoid irradiation. AB - Purified CD4+ BALB/c spleen T cells obtained 4-6 wk after total lymphoid irradiation (TLI) helped normal syngeneic B cells to produce a vigorous antibody response to TNP keyhole limpet hemocyanin in adoptive cell transfer experiments. However, the same cells failed to transfer delayed-type hypersensitivity to the adoptive hosts as measured by a foot pad swelling assay. In addition, purified CD4+ cells from TLI-treated mice were unable to induce graft vs. host disease in lethally irradiated allogeneic C57BL/Ka recipient mice. In response to mitogen stimulation, unfractionated spleen cells obtained from TLI mice secreted normal levels of IL-4 and IL-5, but markedly reduced levels of IL-2 and INF-gamma. A total of 229 CD4+ clones from spleen cells of both normal and TLI-treated mice were established, and the cytokine secretion pattern from each clone was analyzed. The results demonstrate that the ratio of Th1- and Th2-like clones in the spleens of normal BALB/c mice is 1:0.6, whereas the ratio in TLI mice is approximately 1:7. These results suggest that Th2-like cells recover rapidly (at approximately 4-6 wk) after TLI treatment and account for the early return of antibody helper activity and secretion of IL-4 and IL-5, but Th1-like cells recover more slowly (in approximately 3 mo) after irradiation, and this accounts for the deficit in cell-mediated immunity and the reduced amount of IL-2 and IFN gamma secretion. PMID- 2572668 TI - Induction of human IgE synthesis by CD4+ T cell clones. Requirement for interleukin 4 and low molecular weight B cell growth factor. AB - We have analyzed in detail the precise requirements for the induction of human IgE synthesis using several experimental approaches with purified B cells and well-characterized alloantigen-specific CD4+ T cell clones expressing different profiles of lymphokine secretion. Using these clones under cognate conditions in which the B cells expressed alloantigens recognized by the cloned T cells, we have confirmed that IL-4 is required for the induction of IgE synthesis, but we have clearly demonstrated that IL-4 by itself is not sufficient. With several cloned CD4+ T cell lines, including an IL-4-producing clone that could not induce IgE synthesis, and cloned T cells pretreated with cyclosporin A to inhibit lymphokine synthesis, we showed that Th cell-B cell interactions are necessary for IgE synthesis, and that low molecular weight B cell growth factor (LMW-BCGF) and IL-4, in combination, are lymphokines of major importance in the induction of IgE synthesis. Together our results indicate that optimal induction of an IgE specific response requires the exposure of B cells to a particular complex of signals that include (a) a signal(s) involving Th-B cell interaction that primes B cells to receive additional signals from soluble lymphokines, (b) a specific B cell proliferative signal provided by LMW-BCGF, and (c) a specific B cell differentiation signal provided by IL-4. PMID- 2572670 TI - Molecular analysis of human gamma/delta+ clones from thymus and peripheral blood. AB - We analyzed the V gamma and V delta gene usage in TCR-gamma/delta-bearing T cell clones isolated from human peripheral blood and postnatal thymus using V-specific mAbs and Southern and Northern analyses. In peripheral blood most of the gamma/delta cells express the V gamma 9-JP-C gamma 1 chain paired with a delta chain bearing the V delta 2 gene product. This heterodimer is very rare in the postnatal thymus, where a different and less restricted pairing of V gamma 9 and V delta 2 chains is found. These findings indicate that physical constraints cannot explain the overrepresentation of a particular V gamma 9-JP/V delta 2 heterodimer in the peripheral blood, and we discuss alternative mechanisms that may account for this differential distribution. In addition, this analysis allowed us to map the specificity of the delta TCS1 mAb to V delta 1-J delta 1 and to identify at least five different expressed V delta genes. PMID- 2572671 TI - Intraepithelial lymphocytes. Anatomical site, not T cell receptor form, dictates phenotype and function. AB - The function and structure of the TCR proteins of intraepithelial lymphocytes (IEL) were examined using a panel of mAbs specific for TCR-gamma/delta. Three subsets of TCR-gamma/delta+ IEL could be detected with five mAbs, termed GL1-GL5. The mAbs were able to trigger lysis via crosslinking of the IEL TCR and all of the subsets were constitutively cytolytic. Immunoprecipitation of IEL TCR proteins revealed that the GL2 mAb reacted only with gamma, delta heterodimers containing high Mr delta chains, while the other mAbs precipitated all of the observed gamma and delta proteins. Two-color fluorescence analysis showed that the GL2+ subset was contained within the larger GL1+ subset. The GL3 and GL4 mAbs appear to be specific for all TCR-gamma/delta while GL2 was V delta 4 specific. Analysis of IEL for TCR-alpha/beta expression demonstrated that approximately 20% of B6 IEL were TCR-alpha/beta+. Interestingly, this population of IEL contained Thy-1- and CT1+ cells, indicating that the unique phenotype of IEL was not restricted to TCR-gamma/delta+ cells. Moreover, the TCR-alpha/beta+ IEL were also constitutively cytolytic, suggesting that the intestinal milieu was controlling the functional programming of IEL regardless of TCR type. The mAbs reported here as well as the ability to exploit the distinct phenotype of IEL should prove useful in determining the function of IEL and the TCR-gamma/delta. PMID- 2572673 TI - Evaluation of deoxyribonucleic acid (DNA) isolated from human bloodstains exposed to ultraviolet light, heat, humidity, and soil contamination. AB - This study was designed to analyze the effects of common environmental insults on the ability to obtain deoxyribonucleic acid (DNA) restriction fragment-length polymorphisms (RFLP) patterns from laboratory prepared specimens. The environmental conditions studied include the exposure of dried bloodstains to varying amounts of relative humidity (0, 33, 67, and 98%), heat (37 degrees C), and ultraviolet light for periods of up to five days. In addition, the effect of drying over a four-day period in whole blood collected with and without ethylenediaminetetraacetate (EDTA) was examined. The results of the study showed that, under the conditions studied, the integrity of DNA is not altered such that false RFLP patterns are obtained. The only effect observed was that the overall RFLP pattern becomes weaker, but individual RFLP fragments are neither created nor destroyed. PMID- 2572672 TI - Colony morphology of piliated Neisseria meningitidis. AB - An association between piliation and colony morphology has not been observed for the meningococcus. We have found that growth of meningococci overnight at 30 degrees C in a candle extinction jar allows observation of distinct colonial phenotypes correlated to the presence or absence of piliation and the expression of opacity-associated proteins. These phenotypes are similar to those observed in gonococci grown overnight at 37 degrees C. PMID- 2572674 TI - The effects of environment and substrata on deoxyribonucleic acid (DNA): the use of casework samples from New York City. AB - This study was designed to analyze the effects of the environment and substrata on the quality of deoxyribonucleic acid (DNA) isolated from evidentiary specimens. The quality of DNA isolated from actual casework specimens was determined by measuring its size by agarose gel electrophoresis. The information obtained could be used to predict the suitability of the DNA in the samples for restriction fragment length polymorphism (RFLP) analysis. The evidentiary specimens chosen for DNA were classified according to substrate (scrapings, plastic bags, synthetics, denim, and carpet) and according to a subjective evaluation of the condition of the stain (soiled, damp, or putrefied) and to its size (small or large). The results show that DNA of sufficient quality and high molecular weight (HMW) can be reliably isolated from bloodstains deposited on evidentiary items which have an unknown environmental history and which have dried onto a variety of substrata. Subsequent RFLP analysis of a selected number of these samples verified that the DNA was suitable for this type of analysis. PMID- 2572675 TI - Multiple versus single antipsychotic drug treatment in chronic psychosis. AB - Fourteen patients receiving multiple antipsychotic drugs in a state mental hospital long-term unit comprised the study sample. They completed a 1-year clinical trial to reduce such drugs to a single antipsychotic agent. Six of the 14 patients were successfully converted to a single antipsychotic drug without clinical deterioration. Eight patients showed marked psychiatric decompensation when converted to single antipsychotic therapy. Factors that may have contributed to this difference are discussed. PMID- 2572676 TI - Asymmetrical effects of neuroleptics on psychotic patients' performance of a tactile discrimination task. PMID- 2572677 TI - Extrapyramidal symptoms with addition of lithium to neuroleptics. PMID- 2572678 TI - Stimulation of inositol phospholipid hydrolysis by excitatory amino acids is enhanced in brain slices from vulnerable regions after transient global ischemia. AB - Stimulation of inositol phospholipid hydrolysis by transmitter receptor agonists was measured in slices from hippocampus, cerebral cortex, and corpus striatum at various intervals after transient global ischemia in rats. Ischemia was induced through the four-vessel occlusion model. Stimulation of [3H]inositol monophosphate formation by excitatory amino acids was greatly enhanced in hippocampal slices prepared from ischemic rats at 24 h or 7 days after reperfusion. This potentiation was more evident using ibotenic acid and was also observed in cerebral cortex, but not in corpus striatum. This regional profile correlated with the pattern of ischemia-induced neuronal damage observed under our experimental conditions. The enhanced responsiveness to excitatory amino acids was always accompanied by an increase in both basal and norepinephrine stimulated [3H]inositol monophosphate formation. In contrast, stimulation of [3H]inositol monophosphate formation by carbamylcholine was not modified in hippocampal or cortical slices from ischemic animals. PMID- 2572679 TI - Central regulation of adrenal tyrosine hydroxylase: effect of induction on catecholamine levels in the adrenal medulla and plasma. AB - The effect of induction of adrenal tyrosine hydroxylase (TH) by various centrally acting drugs on catecholamine levels in adrenal and plasma was investigated in rats. All the drugs tested, namely oxotremorine, Piribedil, B-HT 920, and HA-966, produced significant increases in adrenal dopamine content and plasma epinephrine level. Denervation of the adrenal abolished the increase in adrenal dopamine as it did the induction of tyrosine hydroxylase. The results suggest that the induced increase of adrenal TH activity, as mediated by certain drugs, results in an elevation of the plasma epinephrine level and that the adrenal dopamine content is a better indicator of the catecholamine-synthesizing capacity of the adrenal medulla than are the other catecholamines. PMID- 2572680 TI - Inhibition of excitatory amino acid-stimulated phosphoinositide hydrolysis in the neonatal rat hippocampus by 2-amino-3-phosphonopropionate. AB - The effects of excitatory amino acid agonists and alpha-amino-omega phosphonocarboxylic acid antagonists on phosphoinositide hydrolysis in hippocampal slices of the 7-day neonatal rat were examined. Significant stimulation of [3H]inositol monophosphate formation was observed with ibotenate, quisqualate, L-glutamate, L-aspartate, alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid, L-homocysteate, and kainate. N-Methyl-D-aspartate had no effect. Of these agonists, ibotenate and quisqualate were the most potent and efficacious. Stimulations by ibotenate and quisqualate were partially inhibited by L-2-amino-4-phosphonobutyrate (10(-3) M), but this antagonist had no effect on L-glutamate, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid, or kainate. At 10(-3) M, D,L-2-amino-3-phosphonopropionate completely inhibited ibotenate and quisqualate stimulations, partially inhibited L-glutamate stimulation, and had no effect on alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid-, kainate-, or carbachol-induced [3H]inositol monophosphate formation. Concentration-effect experiments showed D,L-2-amino-3 phosphonopropionate to be five times more potent as an antagonist of ibotenate stimulated phosphoinositide hydrolysis than L-2-amino-4-phosphonobutyrate. Thus in the neonatal rat hippocampus, like in the adult rat brain, D,L-2-amino-3 phosphonopropionate is a selective and relatively potent inhibitor of excitatory amino acid-stimulated phosphoinositide hydrolysis. Because this glutamate receptor is uniquely sensitive to D,L-2-amino-3-phosphonopropionate, these studies provide further pharmacological evidence for the existence of a novel excitatory amino acid receptor subtype that is coupled to phosphoinositide hydrolysis in brain. PMID- 2572681 TI - Adenosine receptor activation and the regulation of tyrosine hydroxylase activity in PC12 and PC18 cells. AB - We compared the response of rat PC12 cells and a derivative PC18 cell line to the effects of adenosine receptor agonists, antagonists, and adenine nucleotide metabolizing enzymes. We found that theophylline (an adenosine receptor antagonist), adenosine deaminase, and AMP deaminase all decreased basal cyclic AMP content and tyrosine hydroxylase activity in the PC12 cells, but not in PC18 cells. Both cell lines responded to the addition of 2-chloroadenosine and 5'-N ethylcarboxamidoadenosine, adenosine receptor agonists, by exhibiting an increase in tyrosine hydroxylase activity and cyclic AMP content. The latter finding indicates that both cell lines contained an adenosine receptor linked to adenylate cyclase. We found that the addition of dipyridamole, an inhibitor of adenosine uptake, produced an elevation of cyclic AMP and tyrosine hydroxylase activity in both cell lines. Deoxycoformycin, an inhibitor of adenosine deaminase, failed to alter the levels of cyclic AMP or tyrosine hydroxylase activity. This suggests that uptake was the primary inactivating mechanism of adenosine action in these cells. We conclude that both cell types generated adenine nucleotides which activate the adenosine receptor in an autocrine or paracrine fashion. We found that PC12 cells released ATP in a calcium-dependent process in response to activation of the nicotinic receptor. We also measured the rates of degradation of exogenous ATP, ADP, and AMP by PC12 cells. We found that the rates of metabolism of the former two were at least an order of magnitude greater than that of AMP. Any released ATP would be rapidly metabolized to AMP and then more slowly degraded to adenosine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572682 TI - Glutamate-induced 45Ca2+ uptake into immature cerebral cortex neurons shows a distinct pharmacological profile. AB - Glutamate-induced 45Ca2+ uptake was studied in cerebral cortex neurons cultured for 4 days, i.e., at a developmental stage where the neurons are sensitive to the mixed agonist glutamate but not to the actions of N-methyl-D-aspartate or other excitatory amino acids. Using this experimental approach, allowing the investigation of effects elicited only by glutamate, it was demonstrated that the glutamate-stimulated Ca2+ influx could be completely antagonized by MK-801, phencyclidine, and cyclazocine in the nanomolar range, and by 3-(2 carboxypiperazin-4-yl)propyl-1-phosphonate and D-2-amino-5-phosphonopentanoate (APV) in the low micromolar range. However, the glutamate response was unaffected by variations in the Mg2+ concentration in the exposure media. In addition, the two quinoxalinediones 6-cyano-7-nitroquinoxaline-2,3-dione and 6,7 dinitroquinoxaline-2,3-dione were equipotent with APV in blocking the glutamate stimulated Ca2+ uptake. PK 26124 blocked the response in the high micromolar concentration range. Ketamine and gamma-glutamylaminomethylsulfonate were essentially without effect at concentrations up to 10 microM and 300 microM, respectively. These results may suggest the existence of a glutamate receptor with a pharmacological profile not compatible with the existent classification of glutamate receptor subtypes. PMID- 2572683 TI - Restlessness of the arms as the principal manifestation of neuroleptic-induced akathisia. PMID- 2572684 TI - Expenditures for nonspecific back injuries in the workplace. AB - This study reports results based on the 29,421 Tennessee Workers' Compensation claims that were closed in 1986. The study encompasses over $160 million in paid claims. Nearly 8000 cases (27%) involved back injuries, of which more than 90% were nonspecific. Forty percent of the total expenditures were for these nonspecific back injuries. Counterpart data on 8696 injuries involving extremities are included for comparison. Medical expenses account for about 40% of all payments. Data concerning time intervals between injury and return to work are provided. Separate analyses reveal characteristics of those subjects in the highest total expenditure decile, the 2941 subjects who received 57% of total dollar payments. This report provides fiscal data supporting recent appeals for a reappraisal of the clinical management of nonspecific back problems. PMID- 2572685 TI - c-erbB-2 oncogene product staining in gastric adenocarcinoma. An immunohistochemical study. AB - The c-erbB-2 oncogene has been shown to be amplified in a variety of human adenocarcinomas. Antibodies to the protein product, p185, have been used for immunostaining of paraffin-embedded material, and have demonstrated that high levels of c-erbB-2 protein expression correlate with gene amplification under certain conditions. In studies by others, amplification has been demonstrated in 40 per cent of tubular type adenocarcinomas of the stomach, and an immunohistochemical study on frozen tissue has demonstrated staining in 3 out of 10 cases. Our study, using paraffin-embedded material, demonstrates staining in 19 per cent of 126 cases using a polyclonal antibody. Of the positive cases, 75 per cent were tubular or papillary type (P less than 0.025), and prominent staining was restricted to this group. Three cases showed well-defined positive areas in keeping with clonal expression of p185. No specific staining of normal or dysplastic epithelium adjacent to the carcinomas was found. PMID- 2572686 TI - Amplification of DNA for detection of cystic fibrosis-linked polymorphisms. PMID- 2572687 TI - Sulfasalazine hypersensitivity with hepatotoxicity, thrombocytopenia, and erythroid hypoplasia. AB - Sulfasalazine, a commonly prescribed drug for the treatment of inflammatory bowel disease, can cause an allergic reaction with hepatotoxicity, lymphadenopathy, and rash. In addition, many hematologic complications have been observed with sulfasalazine, including aplastic anemia, thrombocytopenia, and erythroid hypoplasia. However, thrombocytopenia in association with sulfasalazine hypersensitivity has not been previously reported. We report a child who developed severe thrombocytopenia and anemia during a hypersensitivity reaction to sulfasalazine. PMID- 2572688 TI - Drug- or hormone-induced adaptation: model of adrenergic hypersensitivity. AB - A pharmacokinetic/pharmacodynamic model of hypersensitivity to adrenergic stimulation following abrupt withdrawal of chronic beta blockade was developed. The model employs the Hill equation, a term which describes the competition between isoproterenol and l-propranolol for beta receptors, and a kinetic term which characterizes the appearance and disappearance rates of up-regulated beta receptors. The model predicted peak chronotropic hyperresponsiveness to isoproterenol 48 hr following abrupt withdrawal of chronic treatment with daily propranolol doses of 160 mg, and a drug half-life of 3.5 hr. The model also predicted that increasing the dose rate and prolonging the half-life of propranolol delayed and decreased the extent of adrenergic hypersensitivity. The time-course of adrenergic hypersensitivity simulated by our model was in excellent agreement with that observed in studies which were published earlier by our laboratory. The model underestimated the extent of adrenergic hypersensitivity. The results of our simulation are consistent with a beta agonist-receptor-effector system, which involves spare receptors, amplification of response by second and third messengers, and beta agonist- antagonist-induced receptor regulation. PMID- 2572689 TI - D2/D1 ratio in the medial preoptic area affects copulation of male rats. AB - The D1/D2 dopamine agonist apomorphine, microinjected into the medial preoptic area (MPOA), facilitates male rat sexual behavior and the D1/D2 antagonist cis flupenthixol in the MPOA impairs it. The present study investigated the roles of D1 and D2 receptors in the regulation of copulation by microinjecting drugs selective for these receptors into the MPOA. The D2 agonist LY-163502 delayed the onset and slowed the rate of copulation and also reduced the number of vaginal intromissions required to trigger ejaculation (reduced ejaculatory threshold). The D1 agonist SKF-82526 had no effect, either alone or together with LY-163502. The D1 antagonist SCH-23390 delayed the onset of copulation and decreased ejaculatory threshold, as had the D2 agonist. A low dose of the D2 agonist alone and together with the D1 antagonist delayed the onset of copulation and reduced ejaculatory threshold; the combination of drugs was more effective than LY-163502 alone. Only the combination of drugs slowed the rate of copulation and delayed the resumption of copulation after an ejaculation. Thus, increasing the D2/D1 ratio in the MPOA, by selective stimulation of D2 and/or antagonism of D1 receptors, delays the onset of copulation and reduces ejaculatory threshold, possibly by altering autonomic control of penile reflexes. PMID- 2572690 TI - Analogs of somatostatin selectively label distinct subtypes of somatostatin receptors in rat brain. AB - Somatostatin (SRIF) is a neurotransmitter in the brain. Subtypes of SRIF receptors may mediate the diverse physiological actions of SRIF in the central nervous system. In the present study, the characteristics of subtypes of brain SRIF receptors were examined using two SRIF analogs. [125I]CGP 23996 and [125I]MK 678. [125I]CGP 23996 binds selectively to rat brain SRIF receptors in a saturable manner and with high affinity. [125I]CGP 23996 binding to brain SRIF receptors is inhibited by SRIF agonists with a rank order of potency of SRIF greater than cyclo (aha-Cys-Phe-D-Trp-Lys-Thr-Cys) greater than SMS 201-995 much greater than MK 678 = L-363,301. [125I]MK 678 labels rat brain SRIF receptors which are not detected by low nanomolar concentrations of [125I]CGP 23996. [125I]MK 678 binding to brain membranes is saturable and of high affinity with a Kd of 0.3 nM and a Bmax of 217 fmol/mg of protein in brain and a Kd of 0.17 nM and a Bmax of 211 fmol/mg of protein in anterior pituitary. [125I]MK 678 binding to brain SRIF receptors is blocked selectively by SRIF analogs. SRIF, SRIF 28, D-Trp8 SRIF, SMS 201-995, cyclo (aha-Cys-Phe-D-Trp-Lys-Thr-Cys) and MK 678 have similar potencies to inhibit [125I]MK 678 binding to brain SRIF receptors. The different rank order of potencies of SRIF analogs to inhibit [125I]CGP 23996 and [125I]MK 678 binding to brain SRIF receptors suggests that these radioligands interact with different subtypes of brain SRIF receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572691 TI - Glutamate receptors of the N-methyl-D-aspartate subtype in the myenteric plexus of the guinea pig ileum. AB - The excitatory amino acids L-glutamate and N-methyl-D-aspartate (NMDA) produced contractions of the myenteric plexus-longitudinal muscle preparation of the guinea pig ileum over the concentration range of 3 X 10(-6) to 10(-3) M. The contractile response to L-glutamate and NMDA, but not carbamyl choline, was blocked noncompetitively by 0.6 mM Mg++. In the absence of Mg++, concentration dependent increases in contractile force also were produced by, in order of potency, L-aspartate, L-homocysteate and D-glutamate, but not by quisqualate, kainate or quinolinate. L-Glutamate was competitively antagonized by the selective NMDA receptor antagonists D-2-amino-5-phosphonovalerate and 3-(2 carboxypiperazin-4-yl)propyl-1-phosphonic acid (3 X 10(-6)-3 X 10(-5) M), as well as by the nonselective excitatory amino acid antagonist gamma-D-glutamylglycine (3 X 10(-4) M). Glutamic acid diethyl ester (3 X 10(-4) M) noncompetitively antagonized L-glutamate. L-Glutamate was not blocked by gamma-D glutamylaminomethyl sulphonate (3 X 10(-4) M), an antagonist which preferentially antagonizes kainate and quisqualate. In addition, the phencyclidine-like drugs etoxadrol (10(-7)-10(-5) M), dextromethorphan (10(-6)-10(-5) M) and 5-methyl 10,11-dihydroxy-5H-dibenzo(a,d)cyclohepten-5,10-imine (10(-9)-10(-7) M) noncompetitively antagonized L-glutamate. The (+) isomer of 5-methyl-10, 11 dihydroxy-5H-dibenzo(a,d)cyclohepten-5,10-imine was approximately 10-fold more potent than the (-) isomer in antagonizing L-glutamate. The present results demonstrate that receptors for the excitatory amino acid L-glutamate are present in the guinea pig myenteric plexus and are of the NMDA subtype.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572692 TI - Serotonin depletion potentiates gastric secretory and motor responses to vagal but not peripheral gastric stimulants. AB - Vagal stimulation is known to release gastrointestinal serotonin. The effect of depletion of serotonin stores on vagally stimulated gastric acid secretion and motility was studied in rats. Pretreatment of rats with parachlorophenylalanine (p-CPA) did not alter basal gastric acid and serotonin secretion but produced a 57% reduction in the intraluminal gastric release of serotonin and a 43 to 100% potentiation of the gastric acid secretory response elicited by intracisternal injection of the stable thyrotropin-releasing hormone analog, RX 77368, in conscious pylorus-ligated rats or urethane-anesthetized rats with an acute gastric fistula. Dose-response studies revealed that the gastric acid secretion induced by submaximal but not high doses of RX 77368 was elevated significantly by p-CPA pretreatment. p-CPA also enhanced the gastric acid output produced by submaximal, but not high doses of the vagal stimulant baclofen, [beta-(p chlorophenyl)-gamma-aminobutyric acid]. In contrast, p-CPA pretreatment had no effect on gastric acid secretion stimulated by bethanechol, histamine or pentagastrin. Selective depletion of central serotonin stores by pretreatment with the neurotoxin 5,7-dihydroxytryptamine given alone, or combined with parachloroamphetamine did not alter RX 77368-stimulated gastric acid secretion. In addition, gastric contractility stimulated by intracisternal injection of RX 77368 was significantly enhanced by p-CPA but not by 5,7-dihydroxytryptamine pretreatment, whereas the contractile response to carbachol was not altered by p CPA pretreatment. These results suggest that depletion of peripheral but not central serotonergic stores potentiate gastric acid secretion and contractility induced by vagally, but not peripherally acting gastric stimulants. Thus, peripheral serotonin may exert an inhibitory tone on vagally stimulated gastric acid secretion and motility in the rat. PMID- 2572693 TI - Celiprolol does not protect against ventricular tachycardia or sudden death in the conscious canine: a comparison with pindolol in assessing the role of intrinsic sympathomimetic activity. AB - The antiarrhythmic and antifibrillatory effects of the beta-1 adrenoceptor antagonist celiprolol were evaluated in a chronic canine model of myocardial infarction and sudden death. Programmed electrical stimulation (PES) was performed in conscious animals 3 to 5 days after a 2-hr occlusion/reperfusion of the left anterior descending coronary artery. Dogs in which PES resulted in a reproducible nonsustaining or sustained ventricular tachycardia (VT) were randomized to receive i.v. celiprolol (3 mg/kg, n = 10) or vehicle (n = 10). PES and measurement of electrophysiologic (parameters were repeated after 30 min and the animals were entered into the sudden death protocol by introducing a 150 microA anodal current to the lumen of the left circumflex coronary artery via a surgically implanted silver wire electrode. Celiprolol failed to prevent the induction of VT, and the outcome of the sudden death protocol did not differ from vehicle with respect to either sudden (within 1 hr of ischemia) or delayed (greater than 1 hr) mortality. VT cycle length and ventricular refractoriness were prolonged (P less than .05) by celiprolol, but other electrophysiologic parameters were unaffected. Heart rate was not altered after drug, but celiprolol antagonized the ischemia-induced increase in rate seen in the vehicle group. Similar electrophysiologic results and mortality data were apparent in a third group of dogs which received pindolol (0.09 mg/kg, n = 8). The failure of both drugs to protect against ischemic ventricular fibrillation in a model in which beta adrenoceptor antagonism has previously proved beneficial may be due in part to related cardiostimulant properties shared by celiprolol and pindolol. PMID- 2572694 TI - Effects of nebivolol, atenolol and propranolol on in vivo cardiovascular and metabolic responses to isoproterenol in dogs. AB - Pentobarbital-anesthetized dogs were administered either dl-propranolol, atenolol or dl-nebivolol at cumulative doses of 0 (vehicle), 0.0025, 0.01, 0.04, 0.16 and 0.64 mg/kg i.v. After each dose, heart rate and diastolic blood pressure responses to isoproterenol (0.125 micrograms/kg/min i.v. for 5 min), as well as plasma glucose, insulin, lactate and free fatty acid responses, were measured. Heart rate and free fatty acid changes were taken as beta-1 adrenergic indices, with the other parameters taken as beta-2 adrenergic indices. The antagonist dose estimated to cause 50% inhibition of each isoproterenol response (ID50) was calculated. Nebivolol and atenolol had nearly identical cardiovascular profiles, which were much more beta-1 selective than that of propranolol [heart rate and blood pressure ID50 values, mg/kg: 0.034, 0.036 (propranolol); 0.058, 0.713 (nebivolol); 0.047, 0.506 (atenolol)]. Propranolol also potently inhibited isoproterenol-stimulated glucose, insulin and lactate increases (ID50S: 0.020, 0.078 and 0.007 mg/kg, respectively). Nebivolol and atenolol were much weaker inhibitors of these metabolic responses than propranolol (5-fold-35-fold and 8 fold-greater than 90-fold, respectively). Insulin responses were equivalently inhibited by both nebivolol and atenolol (ID50S greater than 0.4 mg/kg), whereas glucose and lactate ID50S for nebivolol were 0.183 and 0.243 mg/kg, respectively, with atenolol ID50S greater than 0.64 mg/kg. Free fatty acid responses were attenuated by all three antagonists with ID50 values of 0.103, 0.100 and 0.028 mg/kg for propranolol, nebivolol and atenolol, respectively. These in vivo studies demonstrate that dl-nebivolol significantly inhibited the beta-1 cardiac response at doses which did not produce either beta-2 cardiovascular or metabolic effects. PMID- 2572695 TI - The seating of one-piece and soldered fixed partial dentures. AB - This laboratory study compared the seating of one-piece castings with that of soldered fixed partial dentures. Two investments were investigated, five American Dental Association type III and five metal ceramic alloy fixed partial dentures were made by using the conventional indirect technique. The investing and casting protocol was designed to produce an ideal fit of the individual complete crown abutments, not the fixed partial denture. Independent seating of each retainer provided a baseline measurement for complete seating and was compared with fixed partial denture seating. Fixed partial dentures were then sectioned and reassembled by soldering twice, using a low- and high-expansion investment. Statistical evaluation identified significant differences between the metals and the techniques. Marginal discrepancies with one-piece casting and lower (0.6%) thermal-expansion soldering were clinically unacceptable. A clinically acceptable fit was obtained with the higher (1%) thermal-expansion soldering investment. PMID- 2572696 TI - [Urinary calculi caused by glaphenine. Role of radiology in diagnosis. Apropos of a case]. AB - The authors report a case of urinary calculi secondary to glafenine which posed a difficult problem in the radiological diagnosis. They review the literature of iatrogenic urinary calculi and discuss the place of radiology (Excretory Urography, Ultrasound and Computed Tomography) in the diagnosis. PMID- 2572697 TI - Multiple percutaneous transluminal angioplasties and low dose pulse methotrexate for Takayasu's arteritis. AB - A 20-year-old woman developed left shoulder and arm claudication and angiographic evidence of Takayasu's arteritis. She obtained temporary relief from percutaneous transluminal angioplasties 6 times. Her symptoms were finally controlled with low dose oral pulse methotrexate. This is the first case report of Takayasu's arteritis treated successfully with low-dose pulse methotrexate. PMID- 2572698 TI - Remodeling the pyramid--a concept whose time has not yet come. PMID- 2572699 TI - Use of auditory evoked responses as a measure of recovery from benzodiazepine sedation. AB - The amplitude of the P300 component of auditory evoked responses was found to be depressed by benzodiazepine sedation and was subsequently used to monitor the recovery of volunteers sedated with midazolam. The amplitude of the evoked responses was found to be highly correlated with blood midazolam levels but to be no more sensitive than standard psychomotor testing in assessing recovery from sedation. PMID- 2572701 TI - Rapidly progressive aortic incompetence and coronary artery disease in a patient with Takayasu's disease. PMID- 2572700 TI - Neuroleptics and psychic indifference: a review. PMID- 2572702 TI - Expression of c-myc, c-Ha-ras1, and c-erbB-2 proto-oncogenes in normal and malignant human breast epithelial cells. AB - Short-term cultures of normal human mammary epithelial cells were used to determine the extent to which c-myc, c-Ha-ras1, and c-erbB-2 proto-oncogenes were expressed in proliferating normal cells. This level of expression was compared with that of primary tumor cells, malignant effusion cells, or permanently established breast cancer cell lines. Pure preparations of epithelial organoids from seven different reduction mammoplasty tissue samples yielded proliferating normal epithelial cells upon short-term tissue culture. In every sample, proto oncogene transcript levels increased upon short-term culture of the epithelial cells. These levels often exceeded by 10-fold the levels measured in uncultured organoids from the same tissue. In four of the seven cultured normal breast samples, at least one of the proto-oncogenes increased its expression to a level equaling or exceeding that found in a proliferating breast cancer cell line, MCF7. One effusion metastasis sample and two primary ductal adenocarcinomas were also examined for proto-oncogene expression. The effusion metastasis sample expressed high levels of c-erbB-2 messenger RNA, in accord with its amplified gene copy number; otherwise, the levels of proto-oncogene transcripts were low in unprocessed tumor and uncultured organoids, but they increased with proliferation of the tumor cells in culture. These results indicate that the variable expression of these proto-oncogenes observed in breast biopsy specimens needs to be controlled for cellular growth rate or proliferation index. Furthermore, these findings suggest that dysregulated proto-oncogene expression, rather than overexpression per se, needs to be evaluated as a possible mechanism contributing to the development of human breast cancer. PMID- 2572703 TI - P-glycoprotein expression and AIDS-related Kaposi's sarcoma. PMID- 2572705 TI - Effects of hantaviral infection on survival, growth and fertility in wild rat (Rattus norvegicus) populations of Baltimore, Maryland. AB - Survival, growth rates, body size and fertility of wild caught Norway rats (Rattus norvegicus), infected and uninfected with a Hantavirus (antigenically related to Seoul virus), were compared. No differences were found in the survival of seronegative versus seropositive rats, as measured by mark-recapture experiments. Growth rates, as measured by weight gain but not by increased body length, were slower in seropositive, sexually mature (greater than 200 g) rats, although no differences in the ultimate body size of infected versus uninfected rats were found. No differences in external measures of sexual maturity, or in embryo counts or testes sizes, were found for infected versus uninfected rats. We conclude that hantaviral infections have little or no impact on demographic processes in Norway rat populations. PMID- 2572706 TI - Medical care in the nursing home. PMID- 2572707 TI - From the Alcohol, Drug Abuse, and Mental Health Administration. PMID- 2572704 TI - Induction of c-fos and TIS genes in cultured rat astrocytes by neurotransmitters. AB - The interaction of neurotransmitters with their specific receptors initiates a cascade of intracellular biochemical events which lead to induction of specific genes. Included in this cascade is the rapid and transient induction of a family of primary early response genes we term TIS genes (Lim et al.: Oncogene 1: 263 270, 1987). Expression of six TIS gene, including c-fos, was examined in secondary cultures of rat neocortical astrocytes exposed to muscarinic and adrenergic agonists and antagonists to study the early genomic responses which accompany neurotransmitter-induced alteration of glial morphology and physiology. Carbachol induced accumulation of mRNA for c-fos and the other TIS genes. Carbachol-mediated induction of TIS mRNA expression was sensitive to atropine blockade and was potentiated by lithium. Norepinephrine (NE), isoproterenol, or phenylephrine also induced TIS mRNA accumulation. In order to determine which second-messenger pathways mediate NE induction of TIS gene expression, the influences of the beta(B) antagonist propranolol (PR), the alpha I(AI) antagonist prazosin (PZ), and the alpha 2(A2) antagonist yohimbine (YB) were examined. The induction of TIS1 mRNA by NE was partially blocked by PR or PZ alone, and completely abolished by both antagonists in combination. YB had no effect on TIS1 mRNA expression. These results suggest that NE induces TIS1 mRNA through both B- and A 1-adrenergic, but not A2, pathways. The lack of effect of inhibitors of phospholipase A2 and cyclooxygenase suggests that the A1 component is mediated through a protein kinase C pathway. The induction of transient gene expression by neurotransmitters may mediate the secondary genomic responses and phenotypic changes occurring in astrocytes in response to alterations in neuronal neurotransmitter release. PMID- 2572708 TI - Intraarterial digital subtraction angiography for evaluation of internal mammary artery graft in coronary bypass surgery. AB - Digital subtraction angiography (DSA) was performed to evaluate graft patency in 60 patients who had undergone coronary artery bypass grafting with the internal mammary artery (IMA). Sixty IMA grafts with 62 distal anastomoses, and 84 saphenous vein grafts with 85 distal anastomoses were evaluated by intraarterial DSA with an image frequency of 4 frames/sec. A No. 4 Fr. angiography catheter was introduced percutaneously into the left brachial artery and contrast medium was injected both into the subclavian artery for opacification of the IMA grafts and into the ascending aorta for opacification of aortocoronary saphenous vein grafts. Bypass grafts were defined as patent when grafts were opacified through their entire course, including anastomotic sites. Fifty-nine IMA grafts with 61 distal anastomoses (98%) and 76 saphenous vein grafts with 77 distal anastomoses (91%) were patent. The overall patency rate for total grafts was 94%. In 4 patients, DSA showed a "string sign" in the IMA graft which was patent but narrowed at its distal segment. In 5 patients, large first intercostal branches were opacified. Intraarterial DSA with the retrograde brachial approach is not difficult, and is an excellent low risk method of evaluating the patency of IMA and saphenous vein grafts. DSA provided information about functional patency as well as anatomical patency of the IMA graft. PMID- 2572709 TI - [The effect of succinylcholine on vecuronium and pancuronium]. AB - In 105 adult patients under halothane anesthesia, the neuromuscular blocking effects of vecuronium and pancuronium were determined with prior succinylcholine 1 mg.kg-1 administration and without. Force of the evoked twitch increased 123.7% of control after recovery from succinylcholine-induced block. Prior administration of succinylcholine was associated with a leftward shift of dose response curve of vecuronium or pancuronium. Onset of the force reduction from initial dose (0.08 mg.kg-1) was faster and recovery from initial and maintenance doses (0.02 mg.kg-1) were slower. This potentiating effect persisted at least 2 hours. PMID- 2572711 TI - Variability of DNA fingerprint in a Japanese population. AB - To estimate the forensic usefulness of the DNA fingerprint produced by a synthesized minisatellite probe "Myo", the variability of the DNA fingerprint was examined in a Japanese population. Pairwise comparisons of the fragments in the DNA fingerprints showed that 15 fragments on average longer than 4.4 kilobases with a mean fragment sharing probability of 0.19 appeared in a typical DNA fingerprint and the distribution of matches of the fragments was found to be binominal. Therefore, the probability of chance association between random individuals can be calculated to (0.19)15 = 1.5 x 10(-11). It seems reasonable to conclude that the DNA fingerprint is completely individual-specific. PMID- 2572710 TI - Cardiovascular effects of methylflavonolamine hydrochloride (SIPI-549) in cross circulated canine atrial and ventricular preparations. AB - The cardiovascular effects of SIPI-549 [4'-methyl-7-(2-hydroxy-3 isopropylaminoproxy)-flavone hydrochloride] were investigated in isolated canine atrial and ventricular muscles perfused with heparinized arterial blood from a donor dog. When SIPI-549 was administered intravenously to the intact donor dog, a slight hypotension was induced in a dose range of 0.01 to 3 mumol/kg. At the same time, slight negative chronotropic and inotropic responses appeared in isolated and perfused atria, suggesting a cardiac depressant action. Direct administration of SIPI-549 into the cannulated sinus node artery of the isolated atrium produced negative inotropic and biphasic chronotropic responses in a dose related manner (0.01-1 mumol). SIPI-549 also induced a dose-dependent decrease in developed tension of the isolated left ventricle. The SIPI-549-induced negative chronotropic and inotropic effects were not modified by atropine in doses which blocked carbachol-induced negative chronotropic and inotropic effects. The SIPI 549-induced positive chronotropic effects were inhibited by propranolol but significantly potentiated by imipramine treatment, indicating that a large dose of SIPI-549 produces a release of catecholamine via a non-tyramine like action. Large doses of SIPI-549 did not significantly affect norepinephrine-induced positive chronotropic effects. These results indicate that SIPI-549 may have non cholinergic cardiac depressant properties and no beta-adrenoceptor blocking activity. However, it appears to elicit a release of catecholamines in a non tyramine-like manner at relatively large doses. PMID- 2572712 TI - [Family study on mitochondrial DNA polymorphism]. AB - Mitochondrial DNA restriction fragment length polymorphisms (mtDNA RFLPs) were surveyed among 19 Japanese families with 67 individuals including twins. A mother and children, and siblings of each family showed the same pattern of mtDNA RFLPs. Samples from Japanese war orphans remained in China and their probable relatives were also surveyed and one pair indicating a high probability to be siblings using other genetic markers possessed the same variant of mtDNA. Moreover mtDNA RFLPs was applied for maternity test among two families. The data suggested that parents mistook their children for each other at birth. PMID- 2572714 TI - Influence of cigarette smoking on recurrence of duodenal ulcer. AB - The aim of this study of 225 duodenal ulcer patients was to determine the association of cigarette smoking to ulcer recurrence. In this study, smokers were mostly men whose smoking habit was positively associated with a past history of duodenal ulcer, stress and alcohol consumption. These characteristics are regarded to favor ulcer recurrence. In 161 patients not on maintenance therapy, smokers had a higher recurrence rate than nonsmokers, even when foregoing patients characteristics were taken into consideration. No difference in recurrence rates could be observed between nonsmokers and smokers in 64 patients under maintenance therapy. In conclusion, our findings suggest that smoking plays a major role on recurrence of duodenal ulcer and that maintenance therapy can overcome the adverse effect of cigarette smoking on ulcer recurrence. PMID- 2572713 TI - Depressed natural killer (NK) function in blood and marrow is related to the decrease in CD 11+ cells in acute leukemia. AB - The natural killer (NK) cell activity of the blood and marrow was studied in patients with acute myelogenous leukemia (AML). NK activity of the cells from blood and marrow was significantly decreased in AML to three target cell lines. Both binding and killing capacities of the effector cells were deeply depressed in the blood as well as in the marrow at single cell assay. Surface phenotypic analysis showed a significant decrease in CD 11+ cell subsets, but not in CD 16+ or Leu-7+ cells, in both blood and marrow cells from AML. A significant decrease of large granular lymphocytes (LGL) was also displayed in these samples at morphological examination. The effector cells from AML patients poorly responded to interferon stimulation in NK cytolysis. Taken together, a decrease in CD 11+ cell population with LGL morphology appeared to be responsible for the impaired NK activity in patients with AML. PMID- 2572715 TI - Granulomatous lung disorders. PMID- 2572717 TI - [Adrenergic receptors in the cardiovascular system (their topography, function and regulation)]. PMID- 2572716 TI - Detection and characterization of 3H-clonidine binding sites in coated vesicles isolated from bovine brain. AB - The binding of 3H-clonidine to bovine brain coated vesicles was studied. The binding was reversible and saturable. Saturation studies revealed a one-site interaction: Kd was 8.7 nM and Bmax was 24.5 fmol/mg protein. Noradrenaline, yohimbine and phenoxybenzamine displaced 3H-clonidine binding from the binding sites at concentrations of 10(-7)-10(-4) M. The results indicate that the coated vesicles from the bovine cerebral cortex contain 3H-clonidine binding sites comparable to the low affinity sites of alpha 2-adrenergic receptors in bovine brain membranes. PMID- 2572718 TI - Short-term effects of parathyroidectomy on plasma biochemistry in chronic uremia. AB - Parathyroidectomy (PTx) is indicated in hemodialysis (HD) patients who have severe osteitis fibrosa unresponsive to vitamin D therapy or in whom the latter treatment is contraindicated. Immediately after PTx, plasma immunoreactive parathyroid hormone, calcium and phosphorus concentrations decline abruptly. However, little is known in such patients about the short-term effects of PTx on plasma alkaline phosphatase (AP) activity and plasma aluminum (Al) levels. The present, preliminary study was performed to determine such parameters in 37 HD patients, and to correlate them with data of bone histology. Mean plasma AP activity started to increase after PTx from day 4 onwards. Thus, AP values significantly higher than pre-PTx values were observed at day 7 and 14 (415 +/- 54 vs. 619 +/- 77 and 749 +/- 83 IU/liter, means +/- SEM; N = 37; P less than 0.05 and 0.001, respectively). This increase, in the absence of changes in liver function, was mainly due to the bone-specific iso-AP. Moreover, the degree of increase in plasma AP activity was higher in the subgroup with negative (group I, 21 patients) than in that with positive bone Al staining (group II, 16 patients). However, plasma osteocalcin (BGP) did not change after PTx (N = 8). Basal plasma Al levels were significantly higher in group II both before and two weeks after surgery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572719 TI - Type 1 fimbriate strains of Escherichia coli initiate renal parenchymal scarring. AB - The renal scarring which characterizes chronic pyelonephritis is initiated by bacterial infection and is independent on the activation of an inflammatory response. Although the presence of polymorphonuclear leukocytes (PMN) is essential for initiating the scarring process, the bacterial structures responsible for their activation have not been investigated. In an animal model of chronic pyelonephritis the surface area of the renal scars produced by Type 1 fimbriate escherichia coli (E. coli) was significantly greater than that of those produced by P fimbriate and non-fimbriate strains (P less than 0.01). The activation of human PMN by the same Type 1 fimbriate organisms resulted in a significant release of lysosomal neutral protease activity (P less than 0.001) and activation of the respiratory burst (P less than 0.01). The neutral protease release in response to P fimbriate and non-fimbriate organisms was not significantly increased. The extent of renal scarring also correlated with the release of neutral protease activity (P less than 0.02) and with the degree of activation of the respiratory burst (P less than 0.05). These results demonstrate that the ability of E. coli strains to cause renal scars may be related to their capacity to express Type 1 fimbria, which may be a causative factor in the in vivo activation of the inflammatory response. PMID- 2572720 TI - [Endoscopic electrocoagulation and controlled pharmacological hypotension in the treatment of acute gastrointestinal hemorrhage]. AB - The experience with the treatment of 602 patients with gastrointestinal bleeding before the introduction of endoscopy, and 805--with the use of endoscopy is summarized. At investigation, it was established that in 587 (72.9%) patients, the bleeding stopped, in 180 (27.1%)--it continued, and required in 93 the performance of electrocoagulation hemostasis. In 25 patients, the guided hypotension was used in the complex with the other methods of arresting the bleeding, the favourable outcome was noted in 22. PMID- 2572723 TI - [A case of depression in the treatment of nonspecific ulcerative colitis with sulfasalazine]. PMID- 2572722 TI - [Lipid peroxidation as an indicator of the effectiveness of treating patients with chronic ischemic heart disease using a helium-neon laser]. AB - A therapeutic response to clinical application of Helium-Neon laser irradiation in combination with antianginal drugs was evaluated for chronic sufferers with coronary heart disease (CHD). The largest scope of information on the efficacy of the laser therapy has been provided by the parameters of lipid peroxidation. Individually adjusted doses of the irradiation allowed achievement of the best treatment results. PMID- 2572724 TI - [First International Nursing Research Conference in the German Federal Republic 7 and 8 May, 1989. Nursing research for professional nursing practice]. PMID- 2572721 TI - [Modulation of renal transmitter release by presynaptic receptors]. AB - Renal sympathetic nerve varicosities possess a variety of receptors which when activated by appropriate agonists can modulate noradrenaline release at the local level of the kidney. Thus, activation of prejunctional alpha 1- and alpha 2 adrenoceptors, prostaglandin (PG), dopamine, adenosine and serotonin receptors inhibits, whereas activation of prejunctional beta 2-adrenoceptors and angiotensin (A) II receptors enhances renal noradrenaline release. Moreover, neuronally released noradrenaline itself activates prejunctional inhibitory alpha 1- and alpha 2-adrenoceptors forming a "negative feedback loop" of its own release (autoinhibition). PGE2 and adenosine locally formed in the kidney by renal nerve stimulation inhibits noradrenaline release through activation of their specific prejunctional receptor system (transjunctional inhibition). PMID- 2572726 TI - [Nursing--a new future]. PMID- 2572725 TI - [19th International Congress for nursing of the International Nursing Association. Meeting of the Council of National Representatives from 27 May to 31 May 1989]. PMID- 2572727 TI - [Nursing--a new future. Address on the occasion of the 19th International Congress of the International Council of Nurses]. PMID- 2572728 TI - First Task Force Meeting on Nursing Care of People with AIDS and their Families vom 3.-6. Mai 1989 in Kiel. PMID- 2572729 TI - [What are the effects of beta blockers?]. PMID- 2572731 TI - Factors influencing the calculation of results from studies of the release of tritiated neurotransmitters from superfused slices of guinea pig striata. AB - Slices of guinea pig striata were incubated with tritiated choline, dopamine, or serotonin and the release of radioactive transmitter was studied in a superfusion system. Some experiments were also done on the release of [3H]acetylcholine from rat striatal slices. For analysis of the results, various parameters of the system were determined in order to establish the most reliable method of assessing drug effects on transmitter release. Peaks of radioactivity (S1 and S2) above basal release of radioactivity (B1 and B2) were observed after two depolarizations of the nerve endings with high K+ buffer. Each stimulation was for 2 min, and S2 occurred 20 min after S1. There was a highly significant correlation between S1 and the protein content of the slices for acetylcholine release from guinea pig striata. Basal release of radioactivity, and the ratio S2/S1, were not sensitive to minor changes in the experimental conditions. It was concluded that S2/S1, rather than S1 or S1/B1, should be used as a measure of drug effects on release. The experiments also demonstrated that re-uptake of released neurotransmitter is operative in the superfusion system for dopamine, but not for serotonin. Differences were observed between the rat and the guinea pig with respect to the release of [3H]acetylcholine. PMID- 2572732 TI - Catastrophic consequences of internal mammary artery hypoperfusion. AB - Inappropriate use of the internal mammary artery for complex coronary artery bypass operations may have disastrous yet avoidable consequences. In 712 consecutive coronary artery bypass operations performed between January 1985 and September 1986, five patients had an intraoperative course suggesting internal mammary artery hypoperfusion. In three, coronary artery bypass was performed as a reoperative procedure. The following similarities were noted in all five patients: a technically satisfactory internal mammary artery anastomosis, a left anterior descending coronary artery having critical proximal stenosis (greater than or equal to 90%), a distal artery at least 2.0 mm diameter free of disease, satisfactory left ventricular function, a catastrophic clinical outcome (death in three, cardiac transplantation in one, 3-month hospitalization in one), and all patients could be weaned from cardiopulmonary bypass only after insertion of a saphenous vein graft distal to the internal mammary artery graft. Bilateral internal mammary grafts were used in four of five patients. In both patients without prior coronary artery bypass, there was a discrepancy in internal mammary artery flow and myocardial demand: cardiac hypertrophy, large (2.5 mm) distal left anterior descending supplied by a small (less than 2.0 mm) internal mammary artery, and sequentially grafted arteries. All three patients having reoperative coronary artery bypass had normal anterior wall motion, and in two of the three patients, the left anterior descending-saphenous vein graft was arteriographically normal and the sole source of blood to the anterior, septum, and inferior myocardium. In two patients the normal left anterior descending saphenous vein graft supply a large left anterior descending (greater than 2.0 mm) was replaced with a small left internal mammary artery. Internal mammary artery hypoperfusion typically occurred 30 to 40 minutes after discontinuation of cardiopulmonary bypass but was also seen in the intensive care unit. The condition may be confused with internal mammary artery or coronary artery spasm. It may be avoided by careful prebypass planning of the operation, intraoperative assessment of internal mammary artery flow and size with prudent use of sequential internal mammary artery grafting, avoiding use of the terminal left anterior descending (unless large), retaining nonobstructed saphenous vein grafts when the internal mammary artery has marginal flow or size, and placement of a saphenous vein graft distal to the left internal mammary artery when size or flow in the latter is small. PMID- 2572733 TI - Cardiac surgery in septuagenarians: is there a difference in mortality and morbidity? AB - Open heart surgery is being performed with increased frequency in elderly patients. Results vary considerably probably as the profile of the surgical population changes. A comparison was established by means of univariate analysis between two consecutive series of 100 patients undergoing bypass grafting or valvular replacement. Group A patients were 70 years of age or older; group B patients were less than 65 years of age. Various clinical, operative, and postoperative parameters were monitored. A significant difference was found between the two groups. Elderly patients tended to be white, female, with poor left ventricular and renal function, had carotid or peripheral vascular disease, and new onset of symptoms. Fewer patients had single-vessel disease and received internal mammary artery grafts. Operative mortality rate was higher in group A subjects and postoperative neurologic complications were also more frequent in this group. Factors influencing operative mortality were the association between vascular disease, presence of left main coronary artery disease, and urgency of the procedure. The mortality rate was 17.2% in 29 patients with associated vascular disease and 4.8% (3/63) in those without vascular disease (p = 0.048). Operative mortality was 4.1% without left main coronary artery disease, 15% with left main coronary artery disease (p = 0.083), and 4% in 84 patients having elective operations versus 31.2% in patients having 16 emergency procedures (p = 0.001). Short-term follow-up revealed a higher late mortality rate in elderly subjects and suggested an increased incidence of early pulmonary embolus. Nevertheless, elective cardiac surgery may be safely performed in septuagenarians with an expected operative mortality rate under 5%. PMID- 2572734 TI - An improved method for Lowicryl K4M electron microscopic embedding of brain tissue. AB - The Lowicryl K4M low-temperature embedding technique was modified for brain tissue to facilitate antigen localization in neuronal structures by using postembedding immunoelectron microscopy. Rat brains fixed by perfusion with a mixture of 1% glutaraldehyde and 1% paraformaldehyde, were sectioned into 200 microns-thick coronal sections, and the appropriate area of the sections further microdissected into small samples (200 x 700 x 700 microns). These samples were processed for K4M embedding using prolonged incubation times at each step. This resulted in a homogeneously infiltrated tissue with good preservation of ultrastructure and antigen reactivity. Despite the lack of osmium fixation, the morphology of the synapses was clearly identifiable. PMID- 2572730 TI - Prenatal diagnosis and prevention of inherited abnormalities of collagen. AB - There is now strong evidence for the implication of collagen alpha 1(I), alpha 2(I) and alpha 1(III) mutations in many forms of osteogenesis imperfecta and inherited arterial aneurysms (Ehlers Danlos syndrome type IV). A sizeable proportion of these disorders have detectable abnormalities by conventional protein chemistry, immunofluorescence, or more sophisticated DNA analysis. Everyone of them with specific defects or with linkage to appropriate gene markers is therefore amenable to prevention using conventional prenatal diagnosis by chorionic villus biopsy (with fibroblast culture), fetoscopic biopsy (with fibroblast culture), ultrasound diagnosis of the severely deformed fetus, or gene linkage studies by chorionic villus biopsy or amniocentesis. Already many collagen alpha 1(I), alpha 2(I) and alpha 1(III) mutations have been characterized including point mutations, small and large deletions and regulatory mutations. Many others are likely to be rapidly studied by exploiting recent advances in DNA technology, and other strong candidate genes include collagen II (some chondrodystrophies), collagen VI (certain arterial and cardiovascular diseases) and collagen VII (dystrophic epidermolysis bullosa). Other important common diseases are likely to include osteoporosis, osteoarthritis and cerebral aneurysms. A detailed review is provided of collagen interstitial genes and proteins, together with a description of the various forms of osteogenesis imperfecta and Ehlers Danlos syndrome in which either collagen alpha 1(I), alpha 2(I) or alpha 1(III) mutations have been identified. Appropriate restriction length polymorphisms (RFLPs) useful in identifying carriers of these mutant genes are also described. PMID- 2572735 TI - A method permitting precise trimming of resin-embedded tissue for ultrathin sectioning in pre-embedding immunoelectronmicroscopy. AB - Due to the high complexity of the mammalian central nervous system, sampling of immunohistochemically processed brain tissues for electronmicroscopy requires an accurate and reliable technique. For this reason, the flat-embedding method, which allows light microscopical examination of tissue before sampling, is generally employed. Because of the osmification process, however, the tissue is blackish and opaque which hampers light microscopical selection of tissue areas of interest. We have found that tissue translucency is highly improved by an osmification process using an osmium tetroxide-ferrocyanide mixture. We describe a transilluminated chuck that enables visualization of immunostaining in specimens mounted on a trimming instrument, thus allowing for extremely precise sampling of the tissue. PMID- 2572737 TI - Tremors. AB - In this article, normal tremor and common types of pathologic tremors seen in the elderly are defined and described along with a review of current treatments. Problems of differential diagnosis are emphasized. PMID- 2572736 TI - Modulation of thymosin alpha 1 and thymosin beta 4 levels and peripheral blood mononuclear cell subsets during experimental rhinovirus colds. AB - Cellular immune response to localized upper respiratory viral infection was studied in two groups of healthy volunteers infected with an unnumbered rhinovirus serotype (Hanks). In the first group of 18 volunteers, serum levels of thymosin alpha 1 rose on day 5 following rhinovirus challenge. This observation was confirmed in a second group of 20 normal volunteers inoculated with the same rhinovirus strain; there was a slight increase in thymosin alpha 1 levels on day 3 and a significant rise on day 5 (p less than .001). There was also a significant rise in serum thymosin beta 4 levels on day 5 (p less than .001). Serum cortisol rose in parallel with thymosin alpha 1 on day 5 after rhinovirus inoculation, but there was no direct relationship between individual changes in cortisol and thymosin alpha 1 levels. Enumeration of peripheral blood mononuclear cell subpopulations during rhinovirus infection revealed a significant increase in total lymphocytes on day 5 (p less than .01), but not on day 3. The rise in total lymphocyte count was attributable to an increase in T lymphocytes (CD3+) (p less than .05), cytotoxic/suppressor (CD8+) cells (p less than .01) and natural killer (CD16+ cells) (p less than .05). This is the first report of thymic hormone modulation by a virus, and suggests that local rhinovirus infection of the upper respiratory tract has systemic effects and induces cellular immune responses. PMID- 2572738 TI - [A provocation at a congress on medical ethics. Stop hoping for unlimited medical progress. Establish limits in health care by categories based on democratic rules]. PMID- 2572739 TI - Prevalence of antibodies to hepatitis C virus in Spanish patients with hepatocellular carcinoma and hepatic cirrhosis. AB - The prevalence of antibodies against hepatitis C virus (HCV) was investigated in 96 patients with hepatocellular carcinoma, 106 patients with liver cirrhosis without evidence of cancer, and 177 controls without liver disease. 75% of patients with hepatocellular carcinoma had HCV antibodies (anti-HCV), a significantly higher proportion than that observed in patients with cirrhosis (55.6%), or controls (7.3%). The prevalence of anti-HCV was significantly higher in patients with alcoholic cirrhosis and hepatocellular carcinoma (76%) than in patients with alcoholic cirrhosis alone (38.7%) whereas in patients with cryptogenic cirrhosis there was no significant difference between those with and without primary liver cell cancer (81.4% and 77.5%, respectively). These results indicate that HCV infection may have a role in the pathogenesis of hepatocellular carcinoma, even in patients with chronic liver disease apparently related to other agents such as alcohol, and that this recently identified hepatitis virus may be found in a large proportion of patients with cryptogenic cirrhosis. PMID- 2572740 TI - Prevalence of antibodies to hepatitis C virus in Italian patients with hepatocellular carcinoma. AB - A sensitive radioimmunoassay was used to detect antibodies to hepatitis C virus (HCV) in patients with hepatocellular carcinoma and chronic hepatitis. HCV antibodies (anti-HCV) were detected in 86 of 132 patients with hepatocellular carcinoma with no relation to the presence or absence of hepatitis B surface antigen (HBsAg). The prevalence of anti-HCV was also high in patients with diseases thought to predispose to hepatocellular carcinoma, such as non-A, non-B chronic hepatitis and cirrhosis (74%). In HBsAg-negative patients with hepatocellular carcinoma the prevalence of anti-HCV was lower than that in patients with non-A, non-B chronic hepatitis (16% vs 55%); the prevalence of serum antibodies to hepatitis B core antigen (anti-HBc), a marker of hepatitis B virus infection, was 70% and 28%, respectively. In HBsAg-negative patients with hepatocellular carcinoma, anti-HCV and anti-HBc occurred together nearly three times as often as in patients with chronic hepatitis (54% vs 19%). These data indicate that, in Italy, HCV is an important factor associated with hepatocellular carcinoma and non-A, non-B chronic hepatitis. PMID- 2572742 TI - Mumps meningitis and MMR vaccination. PMID- 2572741 TI - Saturation of fat and cholecystokinin release: implications for pancreatic carcinogenesis. AB - In a study to determine the effect of saturation of fats on their ability to stimulate cholecystokinin (CCK) release six normal volunteers ate five test meals containing different fats with intervals of 1 week. Plasma CCK levels were measured by a specific radioimmunoassay and the gallbladder volume was calculated from ultrasound measurements. The sodium salt of the monounsaturated fatty acid oleic acid (3.5 g) produced a significantly greater integrated CCK response than that of the saturated fatty acid stearic acid (mean [SEM] 103 [41] vs 8[41] pmol.l-1.min). The gallbladder contracted to 42 (3)% of its initial volume after oleate but remained at 89 (8)% of its initial volume after stearate. Integrated CCK responses to dietary triglycerides (30 g) also differed significantly according to the degree of saturation--277 (58) pmol.l-1.min after corn oil (predominantly diunsaturated), 143 (14) pmol.l-1.min after olive oil (predominantly monounsaturated), and 44 (12) pmol.l-1.min after suet (predominantly saturated). The finding that unsaturated fats are stronger stimulants of CCK release than saturated fats may explain the promotion of pancreatic carcinogenesis in rats by unsaturated but not saturated fats and may support the role of CCK in this effect. PMID- 2572744 TI - Concorde remains aloft. PMID- 2572743 TI - Oral iron chelators. PMID- 2572745 TI - Airway obstruction during sleep in children. PMID- 2572746 TI - Campylobacter pylori becomes Helicobacter pylori. PMID- 2572747 TI - Androgen blockade and suppression for prostate carcinoma. PMID- 2572748 TI - Repeated prevalence surveys for monitoring effectiveness of hospital infection control. AB - In a 1400-bedded teaching hospital single-day prevalence surveys of hospital infection were done every six months for 3 years. The prevalence of community acquired infection remained constant; but, after the introduction of a general infection-control policy, the prevalence of hospital-acquired infection (HAI) fell linearly from 10.5% in the second survey to 5.6% in the last. After the introduction of a specific urinary catheter care policy, the prevalence of hospital-acquired urinary tract infection (HAUTI) fell from 3.2% in the first four surveys to 2.0% in the last three. These differences persisted when the results were adjusted by logistic regression for patient risk factors, which varied between surveys: the declines for HAI and HAUTI were then 9.9% to 6.0% and 2.9% to 2.2% respectively. Infection control policies, therefore, can have substantial impact on the prevalence of HAI, and their effectiveness can readily be measured by repeated prevalence surveys. PMID- 2572749 TI - What do western blot indeterminate patterns for human immunodeficiency virus mean in EIA-negative blood donors? AB - To investigate the specificity of western blot indeterminate (WBi) patterns for antibodies to the human immunodeficiency virus type 1 (HIV-1), 100 enzyme immunoassay (EIA) negative donors from whom prospectively obtained recipient sera were available were tested by WB. 20 were WBi, with p24 being the predominant (70%) and generally the only band. Among recipients of WBi blood, 36% were WBi in their 6 month post-transfusion sample, but so were 42% of a control population that had received only WB-negative blood. When serial samples from recipients with a WBi pattern were tested on two occasions, only 35% of results were reproducible. No reciepient of WBi blood became EIA positive, true positive for WB, positive for HIV-1 antigen, or positive for EIA reactivity against recombinant p24 or gp41. The polymerase chain reaction was negative for gag and env HIV-1 sequences in all donors and recipients. Thus WBi patterns are exceedingly common in randomly selected donors and recipients and such patterns do not correlate with the presence of HIV-1 or the transmission of HIV-1 from donor to recipient. PMID- 2572750 TI - Amylinamide, bone conservation, and pancreatic beta cells. AB - Amylinamide is a potent osteoclast-inhibiting peptide that is co-secreted with insulin from the beta cell. It induces profound hypocalcaemia in rats and rabbits and abolishes bone resorption by isolated osteoclasts in vitro. The non-amidated human peptide forms an insoluble fibrillar amyloid deposit that may interfere with beta-cell function and precipitate type II diabetes. PMID- 2572751 TI - Are in-vitro fertilisation and embryo transfer of benefit to all? AB - The efficacy, safety, costs, and benefits of in-vitro fertilisation and embryo transfer (IVF/ET) have been reviewed. IVF/ET benefits only a small proportion of infertile women, it is expensive, and has serious health risks. Therefore policies for the management of infertility in which most financial and manpower resources are applied to prevention of infertility must be developed. Health authorities should also require accreditation of IVF/ET programmes, certification of providers, and ongoing monitoring of clinics. Full and accurate information on IVF/ET must also be disseminated, so that individuals seeking treatment for infertility can make the best informed choice about their care and health authorities can decide the place of IVF/ET in their infertility services. PMID- 2572752 TI - Hyperbaric for carbon monoxide poisoning. PMID- 2572753 TI - Nervous reactions to terfenadine. PMID- 2572754 TI - Prolactin in hypothalamic-hypophysial blood. PMID- 2572755 TI - Stroke prevention and oestrogen replacement. PMID- 2572756 TI - Antibiotics in donated blood. PMID- 2572757 TI - Prenatal diagnosis of junctional epidermolysis bullosa Herlitz type. PMID- 2572758 TI - Fish oil and restenosis rates. PMID- 2572759 TI - Xanthochromia. PMID- 2572760 TI - Dipyridamole plus aspirin. PMID- 2572761 TI - Creased ear lobes. PMID- 2572762 TI - Pregnancy and glomerulonephritis. PMID- 2572763 TI - Clinical signs of dehydration in children. PMID- 2572764 TI - Sampling grossly benign breast biopsy specimens. PMID- 2572765 TI - Innervation of aganglionic intestines of Hirschsprung's disease examined by monoclonal antibody 171B5. PMID- 2572766 TI - Diagnosis of bacterial meningitis. PMID- 2572767 TI - Pernasal vitamin C and the common cold. PMID- 2572768 TI - Respiratory syncytial virus subgroups and pneumonia in children. PMID- 2572769 TI - Ceftazidime for melioidosis. PMID- 2572770 TI - Quinolones and the environment. PMID- 2572771 TI - Obesity as risk factor for acute mountain sickness. PMID- 2572772 TI - Use of glue in treatment of type-B aortic dissections. PMID- 2572773 TI - Heat illness and the armed forces. PMID- 2572774 TI - Improving Third World health. PMID- 2572775 TI - Live donors and hepatic transplantation. PMID- 2572776 TI - Severe ovarian hyperstimulation syndrome and deep venous thrombosis. PMID- 2572777 TI - Tumour necrosis factor and adult respiratory distress syndrome. PMID- 2572779 TI - Continuous clearance of HIV in a vertically infected child. PMID- 2572778 TI - Prognostic usefulness of Ki-67 antigen expression in adult T-cell leukaemia. PMID- 2572780 TI - Renal insufficiency with nebulised pentamidine. PMID- 2572781 TI - Nicotine and Tourette's syndrome. PMID- 2572782 TI - Storage of cerebrospinal fluid on paper. PMID- 2572783 TI - Normal 24-hour variation in a trial natriuretic peptide. PMID- 2572784 TI - Non-invasive phrenic nerve stimulation for intractable hiccups. PMID- 2572785 TI - Non-steroidal anti-inflammatory drugs and the bowel. PMID- 2572786 TI - Effect of perioperative blood transfusion on recurrence of Crohn's disease. PMID- 2572787 TI - Dehydroepiandrosterone and Alzheimer's disease. PMID- 2572788 TI - Urokinase-type plasminogen activator antigen and early relapse in breast cancer. PMID- 2572790 TI - Tumour necrosis factor in familial Mediterranean fever. PMID- 2572789 TI - Increase of endothelial cell growth by sera from diabetic patients with proliferative retinopathy. PMID- 2572791 TI - Propionate production in methylmalonic acidemia. PMID- 2572792 TI - Postoperative monitoring in patients with muscular dystrophy. PMID- 2572793 TI - Effects of endothelium-derived nitric oxide on peripheral arteriolar tone in man. AB - NG monomethyl-L-arginine (L-NMMA), a specific inhibitor of the synthesis of endothelium-derived nitric oxide (NO), was infused into the brachial arteries of healthy volunteers to study the role of NO in the control of forearm blood flow. L-NMMA caused a 50% fall in basal blood flow and attenuated the dilator response to infused acetylcholine but not that to glyceryl trinitrate. These results indicate that the dilator action of endothelium-derived NO contributes to the control of basal and stimulated regional blood flow in man. Impairment of production of NO might account for the abnormalities in vascular reactivity that characterise a wide variety of disease states. PMID- 2572794 TI - Pathogenesis of Crohn's disease: multifocal gastrointestinal infarction. AB - In a prospective study, specimens of resected small and large intestine from fifteen patients with Crohn's disease were prepared by heparin-saline vascular perfusion, followed by either resin casting of the mesenteric vascular supply and tissue maceration or glutaraldehyde perfusion-fixation, resin casting, and tissue clearance. The specimens were examined by macrophotography, histopathology, and either scanning or transmission electronmicroscopy. A pathogenetic sequence of events in Crohn's disease was seen--vascular injury, focal arteritis, fibrin deposition, arterial occlusion mainly at the level of the muscularis propria, followed by tissue infarction or neovascularisation. These features were confined to segments of intestine affected by Crohn's disease and did not occur in normal bowel. The findings suggest that Crohn's disease is mediated by multifocal gastrointestinal infarction. This pathogenetic process is compatible with many of the clinical features of Crohn's disease, and its recognition has important implications for the identification of the primary cause of the illness and advances in clinical management. PMID- 2572795 TI - Association between susceptibility to pre-eclampsia within families and HLA DR4. AB - 56 women who had had proteinuric pre-eclampsia and who had parous sisters were studied. In first pregnancy, proteinuric pre-eclampsia was more common in the sisters than in the maternity hospital population (8/71 [11%] vs 41/1978 [2%]); the relative risk was 6.0. The frequency of HLA DR4 was higher in sisters with pregnancy-induced hypertension than in sisters with normotensive pregnancies (8/18 [44%] vs 10/54 [19%]) and more of them shared HLA DR4 with their spouses (4/14 [29%] vs 0/29). Genetic susceptibility to pre-eclampsia is associated with HLA DR4; it may be conferred by fetomaternal sharing of a single recessive HLA linked gene. PMID- 2572796 TI - Autonomic dysfunction at different ambient temperatures in infants at risk of sudden infant death syndrome. AB - Autonomic function was assessed by measuring the heart rate and blood pressure responses to a change from the horizontal to upright posture at various ambient temperatures in four groups of infants aged 8-12 weeks: 30 infants who had suffered a well-defined unexplained apparent life-threatening event (ALTE); 8 infants who had suffered a cyanotic attack; 24 healthy infants with a sibling who had died from sudden infant death syndrome (SIDS); and 17 healthy infants. Autonomic dysfunction was uncommon in the control group; no infant showed an abnormal heart rate response to postural change (R to R interval maximum/minimum ratio less than 1.0) and a postural fall in blood pressure of greater than 10% occurred in only 1 infant. In contrast, in the ALTE group 9 of 26, 9 of 30, and 4 of 22 infants showed an abnormal heart rate response and 20 of 26, 14 of 30, and 10 of 22 a greater than 10% fall in blood pressure on postural change at 20 degrees C, 25 degrees C, and 30 degrees C, respectively. 1 ALTE infant died of SIDS 14 h after showing an RR max/min ratio of 0.8 and a postural fall in blood pressure of 11% and 14% at 20 degrees C and 30 degrees C. Autonomic function testing should become part of the clinical evaluation of infants at high risk of a sudden unexpected death. PMID- 2572797 TI - Treatment of narcolepsy with L-tyrosine: double-blind placebo-controlled trial. AB - A randomised, double-blind, placebo-controlled study of L-tyrosine was done in ten subjects with narcolepsy and cataplexy. Of twenty-eight visual analogue scales rating mood and arousal, the subjects' ratings in the tyrosine treatment (9 g daily) and placebo periods differed significantly for only three (less tired, less drowsy, more alert). Ratings of daytime drowsiness, cataplexy, sleep paralysis, night-time sleep, overall clinical response, and measurements of multiple sleep latency and tests of speed and attention did not differ significantly between tyrosine and placebo periods. Dietary supplementation with tyrosine 9 g daily for 4 weeks seems to have a mild stimulant action on the central nervous system but this effect is not clinically significant in the treatment of the narcoleptic syndrome. PMID- 2572799 TI - Treatment of right-sided Staphylococcus aureus endocarditis in intravenous drug users with ciprofloxacin and rifampicin. AB - A combination of ciprofloxacin (intravenous then oral) and oral rifampicin was tested in 14 intravenous drug users with right-sided Staphylococcus aureus endocarditis. All 10 patients who completed therapy were cured based on resolution of symptoms and negative blood cultures at 4 weeks post therapy. PMID- 2572800 TI - Multidrug resistance in cancer. PMID- 2572798 TI - Rapid diagnosis of tuberculosis by amplification of mycobacterial DNA in clinical samples. AB - A method based on DNA amplification and hybridisation for the rapid detection of Mycobacterium tuberculosis was used to test 35 clinical specimens (sputum, gastric aspirate, abscess aspirate, biopsy sample) from 34 patients in whom tuberculosis was suspected. M tuberculosis was detected in 15 specimens, 2 of which were negative by standard microbiological criteria (microscopy and/or culture). 20 specimens, negative by standard methods, were also negative by the amplification method. M tuberculosis was also detected in peripheral blood samples of 2 of 4 patients with AIDS from whom the organism had been isolated. PMID- 2572801 TI - Epidural block for Caesarean section and circulatory changes. PMID- 2572802 TI - The selling of patients' data. PMID- 2572803 TI - Neurobiology of incontinence. PMID- 2572804 TI - "Compassionate" release of DDI. PMID- 2572805 TI - Three-dimensional computed tomography. PMID- 2572806 TI - Early infant mortality in West Germany before and after Chernobyl. AB - Early infant mortality rates in West Germany were plotted for regions with different radioactive burdens following the Chernobyl nuclear reactor explosion in April, 1986. In all regions, the logarithms of the mortality rates fitted a linear model between 1975 and 1985, but from May, 1986, immediately after the accident, there was a striking deviation from the model in areas with greatest radioactive fallout. PMID- 2572807 TI - Continuous infusion of octreotide in acromegaly. AB - 15 patients with acromegaly were treated with continuous subcutaneous infusion of octreotide in increasing dose from 200 to 1600 micrograms per 24 h by 200 micrograms increments each week. Patients were studied during the initial 7 h of infusion, weekly at each dose level, then after 1 and 2 months at maximum dosage. 13 patients responded well, as judged by growth hormone (GH) suppression and return of insulin-like growth factor 1 to normal, although 1 patient withdrew due to adverse effects; 2 patients showed no significant reduction in GH. In the 12 responders GH level fell within the first 3 h of infusion at a mean plasma octreotide level of 0.76 (SE 0.26) micrograms/l, corresponding to 25 micrograms of infused drug. Optimum suppression of GH occurred at a dosage of 600 micrograms per 24 h, a plasma drug level of 3.5 (0.5) micrograms/l. A definite reduction in tumour size was seen on computerised tomographic scan in 2 patients. All responders noted subjective improvement in acromegalic symptoms. Adverse effects comprised gastrointestinal disturbances, which were transient and mild in 14 patients but severe in 1; biliary sludging occurred in 1 patient. No significant deterioration in carbohydrate tolerance was seen in the responders. PMID- 2572808 TI - Telecommunication discussion groups for health services and medical research. AB - International university and research institute telecommunication links provide free or low-cost access to several computer networks (commercial and non commercial). Previously used by science researchers, such networks are now used in clinical and social medicine. University affiliation, a personal computer, and a modern and software are needed. A host-computer, at a local computing centre, operates server software that automatically distributes information to subscribers. The server stores all messages in a monthly logbook that can be edited to produce an electronic magazine. The magazine can also be accessed by non-subscribers. The software gives convenient, fast, single-key processing of messages and files and also allows on-line conferences. Discussion groups on several medical specialties and data bases for AIDS and other communicable diseases are already operating. PMID- 2572809 TI - Clostridium difficle infection in health-care workers. PMID- 2572810 TI - Liquid ventilation of preterm baby. PMID- 2572811 TI - Action of nitroprusside in multidrug-resistant hypertension. PMID- 2572812 TI - Bacteriotherapy for clostridium difficile colitis. PMID- 2572813 TI - Carnitine supplementation in pivampicillin treatment. PMID- 2572814 TI - Brain damage and open-heart surgery. PMID- 2572815 TI - Poor metabolisers of debrisoquine reveal their true colours. PMID- 2572816 TI - Lovastatin, isoprenes, and myopathy. PMID- 2572817 TI - Plasma mevalonate response in lovastatin-related myopathy. PMID- 2572818 TI - Future glucose intolerance possibly manifest in youth. PMID- 2572819 TI - Retinal vessel constriction under hyperbaric conditions. PMID- 2572820 TI - Hormone dependency of meningiomas. PMID- 2572821 TI - Rash with amoxycillin-clavulanate therapy in HIV-infected patients. PMID- 2572822 TI - Intralesional vinblastine for oral Kaposi sarcoma in HIV infection. PMID- 2572823 TI - Cytomegalovirus screening in pregnancy. PMID- 2572824 TI - Mycobacterium avium-intracellulare serovars in German AIDS patients. PMID- 2572825 TI - Heatstroke and the Armed Forces. PMID- 2572826 TI - Medical Research Council's plans. PMID- 2572827 TI - Computer-assisted diagnosis in acute abdominal pain. PMID- 2572829 TI - European campaign against cancer. PMID- 2572828 TI - Benzodiazepine prescribing. PMID- 2572830 TI - Myopathy as possible side-effect of cyclosporin. PMID- 2572831 TI - Prenatal diagnosis. PMID- 2572832 TI - Age, vitamin D, and solar ultraviolet. PMID- 2572833 TI - Analgesic abuse and hypertension. PMID- 2572834 TI - Vigabatrin therapy in patient with succinic semialdehyde dehydrogenase deficiency. PMID- 2572835 TI - Animal use at a Milan research institute. PMID- 2572836 TI - Let sleeping giardia lie. PMID- 2572837 TI - Calcium channel autoantibodies in Lambert-Eaton myasthenic syndrome. PMID- 2572838 TI - Monoclonal antibody (HML-1) reactivity of T-cell lymphomas. PMID- 2572839 TI - Detection of vesico-ureteric reflux by indirect radionuclide cystography. PMID- 2572840 TI - Cold-induced injury reaction. PMID- 2572841 TI - Prediction of poor outcome of cutaneous surgery. PMID- 2572842 TI - Salicylic acid and ultraviolet B for psoriasis. PMID- 2572843 TI - Greenhouse effect and renal calculi. PMID- 2572845 TI - Near-death experiences. PMID- 2572844 TI - Lower-limb phlebography in recent paraplegia and tetraplegia. PMID- 2572846 TI - Cathepsin D: an independent prognostic factor for metastasis of breast cancer. AB - 122 patients with primary breast cancer were followed-up for a median of 4.6 years after surgery. The concentration of cathepsin D in tumour cytosol was strongly related to both metastasis-free survival and disease-free survival and was independent of nine conventional prognostic indices. Cathepsin D assay may prove particularly useful in identifying women who, though without lymph node involvement at presentation, are at high risk of metastatic disease. PMID- 2572847 TI - Effects of exercise and fat ingestion on high density lipoprotein production by peripheral tissues. AB - The peripheral production of high density lipoprotein (HDL) cholesterol and of the subclasses HDL2 and HDL3 was assessed by measurement of the arteriovenous fluxes across the human forearm, at rest and after 20 min isometric exercise in the forearm. Eight subjects were studied twice--fasting and after a high-fat meal -and one other subject was studied only after fat loading. In the fasted state the net fluxes of HDL2 and HDL3 cholesterol were slightly negative in the resting forearm, but they became positive during exercise, indicating greater production during short-term muscular activity. The effect of exercise, particularly that on HDL3 cholesterol, was greatly increased by a high-fat meal; the difference in HDL3 cholesterol arteriovenous flux between rest and exercise was significant ( 0.06 [SEM 0.05] vs 0.51 [0.17] mumol/100 ml forearm/min). By contrast, there was no peripheral production of HDL2 or HDL3 cholesterol during exercise in two patients with lipoprotein lipase deficiency. These findings suggest that formation of HDL3 during lipolysis by lipoprotein lipase in the muscle capillary bed is influenced by the supply of chylomicrons and other lipoprotein substrates for this enzyme. Muscle blood flow may therefore be an important determinant of HDL production by this mechanism. The effect of exercise in raising HDL cholesterol, and the inverse relation between exercise and coronary heart disease, may be partly the result of this process. PMID- 2572848 TI - Intercellular adhesion molecule 1 on liver allografts during rejection. AB - The expression of intercellular adhesion molecule 1 (ICAM-1), a ligand for the leucocyte adhesion receptor lymphocyte-function-associated antigen 1 (LFA-1), was studied on liver tissue after transplantation. There was greater ICAM-1 expression on bile ducts, endothelium, and perivenular hepatocytes (structures affected by the rejection process) in patients with acute rejection than in donor livers, patients with stable transplants, or patients with non-rejection complications. The expression on bile ducts and hepatocytes was greater in patients in whom there was progression to chronic, irreversible rejection. In patients with resolving rejection ICAM-1 expression was greatly reduced after high-dose corticosteroid treatment. The expression in patients with non-rejection complications and in those with long-term stable grafts was similar to that seen in the donor controls. The induction of ICAM-1 on tissues may be an important step in the development of the inflammatory response of rejection and in determining which cells are the targets of immune damage. The reduction of ICAM-1 expression seen after successful treatment with high-dose corticosteroids suggests that this might be an important mode of action of these drugs. PMID- 2572849 TI - Detection of residual bcr/abl translocation by polymerase chain reaction in chronic myeloid leukaemia patients after bone-marrow transplantation. AB - The polymerase chain reaction was used to evaluate minimum residual disease in chronic myelogenous leukaemia (CML) patients after bone-marrow transplantation, by amplification of the transcript of the specific bcr/abl hybrid gene. Strict precautions were taken to avoid contamination. Peripheral blood cells from 22 patients transplanted for haematological malignant disorders were analysed. The results were clearcut for positive controls (patients with CML in relapse) and negative controls (patients with malignant disorders other than CML). In 11 of 12 CML patients in clinical and cytogenetic remission the bcr/abl transcript was detected 3 months to 6 years after transplantation. Thus, it appears that cells expressing the bcr/abl mRNA are not eradicated from the blood of CML patients in complete clinical remission even years after bone-marrow transplantation. PMID- 2572850 TI - A neurogenic mechanism for symmetrical arthritis. AB - Human synovium is richly innervated by autonomic and sensory nerve fibres, many of which contain neuropeptides. The hypothesis is that, in addition to a sensory role, some of these fibres modulate the response of the synovial membrane to a variety of noxious stimuli by releasing these peptides. Synovial damage results in acute inflammation in the damaged joint and a neurogenically mediated infiltrate of inflammatory cells in the contralateral joint. These cells might protect the contralateral synovium from injury similar to that in the damaged joint. An increased response would lead to synovitis and symmetrical disease. PMID- 2572851 TI - ACE inhibitors after myocardial infarction. PMID- 2572852 TI - Steroids and croup. PMID- 2572853 TI - Chemotherapy and hepatitis B. PMID- 2572854 TI - Endotoxins in heatstroke. PMID- 2572855 TI - Immature skin. PMID- 2572856 TI - Leucocyte adhesion to cells in immune and inflammatory responses. AB - When activated under physiological or pathological conditions leucocytes transiently adhere to one another or to other cell types such as vascular endothelial cells. This adhesiveness is mediated by specific cell adhesion molecules (CAMs). There are two molecular pathways of leucocyte adhesion--the binding of CD11a-c/CD18 (Leu-CAM family) on leucocytes to CD54 (ICAM-1) on mononuclear leucocytes, fibroblasts, and epithelial and vascular endothelial cells, or of CD2 on T cells to CD58, a cell surface molecule found on a broad range of tissue cells. The adhesion process is essential for leucocyte-mediated cytotoxicity, phagocytosis, chemotaxis, and induction of lymphocyte proliferation and differentiation. It also participates in homing of lymphocytes into lymphoid organs and leucocyte migration from the vascular compartment to extravascular tissues. Leucocyte adhesion is thus a critical step in the development of immune and inflammatory responses. An inherited deficiency of the Leu-CAM family leads to recurrent bacterial infections, which can be fatal in early life, and lack of CAMs on Burkitt's lymphoma cells may enable these cells to escape immunesurveillance. PMID- 2572857 TI - International trial of long-term dexfenfluramine in obesity. AB - In a randomised, placebo-controlled, double-blind study, 822 obese patients of both sexes were given either dexfenfluramine (dF), 15 mg twice daily (404), or placebo (418) in addition to a calorie-restricted diet for 1 year. Patients in both groups lost weight significantly in the first 6 months; after 6 months dF patients had a higher cumulative mean weight loss. Dropout rates were lower in dF patients than in placebo patients, mainly because of dissatisfaction with weight loss in the latter group. More than twice as many dF patients as placebo patients achieved a given weight loss; but more dF patients than placebo patients had transient side-effects (tiredness, diarrhoea, dry mouth, polyuria, and drowsiness). PMID- 2572858 TI - Cancer near potential sites of nuclear installations. AB - Mortality and census data for 400 districts of England and Wales were analysed with respect to existing sites of nuclear power stations and sites where the construction of such installations had been considered or had occurred at a later date (potential sites). Excess mortality due to leukaemia and Hodgkin's disease in young people who lived near potential sites was similar to that in young people who lived near existing sites. Areas near existing and potential sites might share unrecognised risk factors other than environmental radiation pollution. PMID- 2572859 TI - Effect of non-steroidal anti-inflammatory drugs on course of osteoarthritis. PMID- 2572860 TI - Cot deaths and sleeping position. PMID- 2572861 TI - Diarrhoea and malnutrition. PMID- 2572862 TI - Birth in the squatting position. PMID- 2572863 TI - Platelet angiotensin II binding sites in hypertension in pregnancy. PMID- 2572864 TI - Pregnancy loss and fathers with Ehlers-Danlos syndrome. PMID- 2572865 TI - Carbamazepine in bipolar affective disorder. PMID- 2572866 TI - Surgery for small-cell lung cancer. PMID- 2572867 TI - Delusions after extracorporeal shockwave lithotripsy. PMID- 2572868 TI - Neutrophil elastases and haemolytic uraemic syndrome. PMID- 2572869 TI - Erythropoietin and autologous blood donation. PMID- 2572870 TI - NSAIDs and gut damage. PMID- 2572871 TI - The case of the spotted underwear. PMID- 2572872 TI - Aspirin and systemic mastocytosis. PMID- 2572873 TI - Mad dogs, Englishmen, and Scots. PMID- 2572874 TI - Reversal of HIV-phenotype to fulminant replication on macrophages in perinatal transmission. PMID- 2572875 TI - Rapid discrimination between HIV-1 and HIV-2 infection. PMID- 2572876 TI - Multiple sclerosis and macrocytosis. PMID- 2572877 TI - Transmission of HIV: genital ulceration, sexual behaviour, and circumcision. PMID- 2572878 TI - Late abortions. PMID- 2572879 TI - Obstetrics and social change. PMID- 2572880 TI - WCH rather than MCH. PMID- 2572881 TI - Education of alternative practitioners. PMID- 2572882 TI - Homelessness. PMID- 2572883 TI - Role of bronchial musculature. PMID- 2572884 TI - Hymenoptera stings and beta-blockers. PMID- 2572885 TI - Diathermy loop excision and the cervix. PMID- 2572886 TI - Postviral fatigue syndrome and the VP-1 antigen. PMID- 2572887 TI - Non-tuberculous mycobacterial tenosynovitis. PMID- 2572889 TI - Antibodies to hepatitis C virus in children with acute or chronic viral hepatitis. PMID- 2572888 TI - Post-exposure prophylaxis for hepatitis B. PMID- 2572890 TI - L-tryptophan as an adjunct to treatment of bulimia nervosa. PMID- 2572891 TI - Oral contraceptives, breast cancer, and lactation. PMID- 2572892 TI - Coming off long-term nicotine gum. PMID- 2572893 TI - Hormone replacement therapy and breast cancer. PMID- 2572894 TI - Intracoronary infusion of vasoactive peptides. PMID- 2572895 TI - Mitral valve prolapse and infective endocarditis. PMID- 2572896 TI - Blood-films in the diagnosis of malaria. PMID- 2572898 TI - Heatstroke in training: a fatal case in Massachusetts. PMID- 2572897 TI - Oedema protein concentrations in cellulitis and deep vein thrombosis. PMID- 2572899 TI - Comparison of daily and twice-weekly regimens to treat pulmonary tuberculosis. AB - A randomised controlled trial compared the effectiveness and toxicity in pulmonary tuberculosis of two drug regimens containing rifampicin and isoniazid given daily or twice-weekly for 4 months after a 2-month period of intensive treatment with daily isoniazid, rifampicin, and pyrazinamide. 667 patients with newly diagnosed pulmonary tuberculosis were randomly allocated to continue daily treatment with isoniazid (400 mg) and rifampicin (600 mg) or to twice-weekly treatment with isoniazid (900 mg) and rifampicin (600 mg). 544 of the 667 patients (81%) completed the 6-month course (287 of 337 [85%] treated daily and 257 of 330 [79%] treated twice-weekly). Drug toxicity was not a great problem; the treatment was permanently discontinued in only 2% of patients. There was no significant difference at the end of months 5 and/or 6 of chemotherapy between the groups treated daily and twice-weekly in the proportions with bacteriological failure (at least one positive sputum culture with more than 20 colonies) or who had died from tuberculosis (17 [6%] vs 10 [3%]). Nor was there a significant difference in the relapse rate (17 [7%] treated daily vs 10 [4%] treated twice weekly) during follow-up of 12 months. Thus, the twice-weekly regimen was at least as effective as the daily regimen for treatment of pulmonary tuberculosis. PMID- 2572900 TI - Serum 25-hydroxyvitamin D and colon cancer: eight-year prospective study. AB - Blood samples taken in 1974 in Washington County, Maryland, from 25 620 volunteers were used to investigate the relation of serum 25-hydroxyvitamin D (25 OHD) with subsequent risk of getting colon cancer. 34 cases of colon cancer diagnosed between August, 1975, and January, 1983, were matched to 67 controls by age, race, sex, and month blood was taken. Risk of colon cancer was reduced by 75% in the third quintile (27-32 ng/ml) and by 80% in the fourth quintile (33-41 ng/ml) of serum 25-OHD. Risk of getting colon cancer decreased three-fold in people with a serum 25-OHD concentration of 20 ng/ml or more. The results are consistent with a protective effect of serum 25-OHD on colon cancer. PMID- 2572901 TI - Hypocholesterolaemia and increased elimination of low-density lipoproteins in metastatic cancer of the prostate. AB - To study the influence of tumour mass on lipid metabolism, the lipoprotein pattern in untreated patients with newly diagnosed cancer of the prostate was examined. Total cholesterol levels were reduced in patients with evidence of metastasis (n = 30) compared with those without metastasis (n = 73). Since the major fraction of serum cholesterol is contained in low-density lipoproteins (LDL), turnover of LDL was studied in detail in 8 patients compared with 12 age matched healthy men. LDL were cleared faster in the 3 patients with metastatic disease than in the patients without metastasis and in controls, indicating faster catabolism of LDL. Thus in prostatic cancer an increased tumour burden is associated with increased elimination of LDL, which contributes to reduced serum cholesterol levels. PMID- 2572902 TI - Comparison of three intravenous bisphosphonates in cancer-associated hypercalcaemia. AB - Three intravenous bisphosphonates were compared in the treatment of cancer associated hypercalcaemia. 48 patients were randomly allocated to one of three treatment groups (each with 16 subjects)--30 mg pamidronate or 600 mg clodronate, both as single intravenous infusions; or etidronate as three infusions of 7.5 mg/kg per day for three consecutive days. Patients were rehydrated with normal saline before bisphosphonate treatment. All three bisphosphonates lowered serum calcium by inhibiting bone resorption; pamidronate was the most potent in this respect. By comparison with the other groups, more patients in the pamidronate group became normocalcaemic, and the effect on serum calcium was apparent sooner and lasted longer. PMID- 2572903 TI - Plasmodium vivax resistance to chloroquine? AB - Two soldiers continued weekly prophylaxis with 300 mg chloroquine base on their return to Australia from Papua New Guinea but were not protected against Plasmodium vivax malaria. Both had symptoms and parasitaemia although plasma concentrations of chloroquine were considerably higher than those regarded as adequate for suppression of vivax malaria. Parasitaemia did not clear after one of the patients was treated with 600 mg chloroquine base. The results suggest the emergence of strains of P vivax with a reduced susceptibility to chloroquine. PMID- 2572904 TI - HTLV-1 and polymyositis in Jamaica. AB - IgG antibodies to human T-cell lymphotropic virus (HTLV-1) were found in 11 of 13 (85%) Jamaican patients with idiopathic adult polymyositis. The association was first observed in 7 patients with polymyositis who were included in a control group of 100 patients with neurological and neuromuscular diseases in a serological investigation of the prevalence of HTLV-1 antibody in patients with tropical spastic paraparesis. All 7 patients with polymyositis were positive for the antibody by an enzyme-linked immunosorbent assay, confirmed by western blot. Because of this striking association a further 6 patients with polymyositis were identified and tested, 4 of whom were also seropositive for HTLV-1 antibody. PMID- 2572906 TI - Routine diagnostic testing. PMID- 2572905 TI - Meeting the need for female sterilisation. PMID- 2572907 TI - Respiratory infection and sudden infant death. PMID- 2572908 TI - Bat ears without tears. PMID- 2572909 TI - Psychotic symptoms in Alzheimer's disease. PMID- 2572910 TI - Oral hairy leucoplakia. PMID- 2572911 TI - Endosonography: promising method for diagnosis of extrahepatic cholestasis. AB - Endosonography, ultrasonography, and computed tomography (CT) were carried out prospectively in 52 patients with extrahepatic cholestasis. 35 patients had extrahepatic biliary obstructions (21 tumorous, 14 non-tumorous) and 17, with recent gallstone migration within the bile duct, had no extrahepatic obstruction at the time of investigation. The definitive diagnosis was established by surgery (in 39 patients), by transendoscopic sphincterotomy (11 patients), or by retrograde biliary opacification (2 patients). Endosonography was significantly more sensitive than ultrasonography or CT (100% vs 80% and 83%, respectively) in making a positive diagnosis of obstruction. Endosonography was also significantly more accurate than ultrasonography or CT (97% vs 49% and 66%) in diagnosing the cause of the obstruction and more effective in the assessment of the locoregional spread of tumorous obstructions (75% vs 38% and 62%). Thus, endosonography was superior to ultrasonography and CT in the diagnosis and staging of biliary obstructions. PMID- 2572912 TI - Concomitant administration of cyclosporin and ketoconazole in renal transplant recipients. AB - 18 renal transplant recipients receiving cyclosporin, prednisone, and azathioprine were given ketoconazole, a potent inhibitor of the cytochrome P-450 enzyme system. Within a month ketoconazole-induced blockade of cyclosporin metabolism allowed a significant reduction (451 vs 106 mg/day; 77%) of the mean dose of cyclosporin without altering cyclosporin whole blood trough levels, although maximum blood levels were almost halved. This dose reduction was maintained in patients followed up for up to 13 months. Renal and hepatic function were unchanged after the addition of ketoconazole. This drug interaction has the potential to reduce dramatically expenditure on cyclosporin in transplant recipients. PMID- 2572914 TI - Psychological support for cancer patients. PMID- 2572913 TI - Factors associated with hypothermia in patients admitted to a group of inner city hospitals. AB - In a case-control study of factors contributing to hypothermia all fourteen patients (mean age 80 years) admitted to hospital with hypothermia after being found ill indoors also had some other serious illness. They were more likely than control patients to have been alone when taken ill (93 vs 39% of controls), to live alone (86 vs 43%), and to have been found on the floor (79 vs 14%). They were less likely to have been wearing more than indoor clothing (0 vs 50%), or to have had heating on when found (50 vs 89%), but 93% of patients in both groups had heating available. Healthy young adult volunteers who lay immobile on the ground in air at 5 degrees C lightly clothed cooled progressively by 0.57 degrees C (SD 0.32) (rectal T degrees) in 90 min despite doubling of metabolic rate. With better insulation in bed, core temperature stabilised within 90 min, and when they were in an armchair it fell slowly, with no increase in metabolic rate in either case. The findings suggest that hypothermia indoors resulted largely from collapse due to illness when the patient was alone lightly clothed and not in bed. Eight hypothermic patients found outside (in December and January) were younger (mean age 60 years) than the fourteen found indoors; six of these were chronic alcoholics or acutely intoxicated, and six lacked, or had wandered from, a fixed home. PMID- 2572915 TI - Genes and carcinogens. PMID- 2572916 TI - Nasopharyngeal carcinoma, salted fish, and Polynesians. PMID- 2572917 TI - Xamoterol and the failing heart. PMID- 2572918 TI - Low-dose aspirin and twin pregnancy. PMID- 2572919 TI - Juvenile onset diabetes and schizophrenia? PMID- 2572920 TI - Agranulocytosis and dipyrone. PMID- 2572921 TI - Fenoterol by metered dose inhaler and delay of steroid therapy. PMID- 2572922 TI - Delayed onset of vasculitis following isotretinoin. PMID- 2572923 TI - Prenatal exposure to polychlorinated biphenyls. PMID- 2572924 TI - Vigabatrin and urinary 5-oxoproline. PMID- 2572925 TI - Slow recovery from severe foodborne botulism. PMID- 2572926 TI - Cluster of listeriosis isolates with different serovar and phagovar characteristics. PMID- 2572927 TI - Oral limaprost for Raynaud's phenomenon. PMID- 2572928 TI - Monitoring of rheumatoid arthritis. PMID- 2572929 TI - Post-abortion chlamydial pelvic infection. PMID- 2572930 TI - Mature entrants to medicine. PMID- 2572932 TI - Assessing workload in neonatal medicine. PMID- 2572931 TI - Recommended dietary allowances for Europe. PMID- 2572934 TI - Hepatitis C virus infection and liver transplantation. PMID- 2572933 TI - Ethics of screening for hepatitis C virus. PMID- 2572935 TI - Identification of Lassa fever virus infection with recombinant nucleocapsid protein antigen. PMID- 2572936 TI - Epidemic visceral leishmaniasis in southern Sudan. PMID- 2572937 TI - Generalised seizures induced by transcranial magnetic stimulation of motor cortex. PMID- 2572938 TI - Usefulness of tip cultures to detect pulmonary artery catheter-related infections. PMID- 2572939 TI - Phenytoin in pre-eclampsia. PMID- 2572940 TI - HIV infection among sub-Saharan African patients in the UK. PMID- 2572942 TI - HTLV-I infection in western equatorial Africa. PMID- 2572941 TI - Inhibition of HBV DNA replication by ganciclovir in patients with AIDS. PMID- 2572943 TI - Long-term metabolic control and diabetic retinopathy. PMID- 2572944 TI - HIV and the epidemiologist. PMID- 2572945 TI - Another British soldier dies from heat illness. PMID- 2572946 TI - AIDS and the law. PMID- 2572947 TI - Health in England and Wales, 1988. PMID- 2572948 TI - No compensation for HIV-infected haemophiliacs. PMID- 2572949 TI - [Have indications for preventing stress ulcer been changed in relation to aspiration pneumonia?]. PMID- 2572950 TI - Effect of pulse duration on microsecond-domain laser lithotripsy. AB - The pulsed dye laser (wavelength 504 nm, pulse duration 1 microsecond) is widely used for fragmenting urinary and biliary calculi. In this study, the performance of this laser was compared with pulsed dye lasers producing pulse durations of 8 and 20 microseconds. Fragmentation thresholds and fragmentation rates were measured using a variety of urinary and biliary calculi. Effective fragmentation of urinary and biliary calculi was obtained with 1-microsecond and 8-microseconds pulse durations, but satisfactory fragmentation could not be achieved at 20 microseconds. PMID- 2572951 TI - Abnormal signal transduction: a hypothetical model for bipolar affective disorder. AB - The molecular basis of bipolar affective disorder is poorly understood at this time. The episodic nature of the condition in which relatively euthymic periods of variable duration separate periods of mania and depression, and the specificity of lithium therapy suggests that a molecular target of the illness may be a system that bidirectionally influences neurotransmission and is affected by lithium. Signal transduction pathways, which are important mediators of neurotransmitter generated signals, may represent such a system because they: 1) generate second messenger molecules that stimulate neurotransmission and also mediate negative feedback mechanisms, and 2) appear to be a direct target of lithium's action on cells. In this paper, we present a model in which abnormal regulation of signal transduction could lead to the episodic accumulation of biologically active transducers or second messengers. These alterations may result in prolonged effector stimulation which may underlie mania, followed by excessive receptor desensitization, which may result in depression. Using our model we suggest a plausible hypothesis that can explain the clinical spectrum of the disorder and the therapeutic action of lithium. PMID- 2572953 TI - The regulatory role of calmodulin-dependent guanylate cyclase in association with hormonal imprinting in Tetrahymena. AB - The primary interaction with insulin accounted for considerable increases in both the calmodulin content and guanylate cyclase activity of Tetrahymena. Both activities were still elevated after 24 h (6-8 generations), but while the calmodulin level showed a decrease, guanylate cyclase activity showed a further significant increase relative to the immediate response. A second treatment with insulin decreased rather than increased both activities, but to dissimilar degrees, in that the calmodulin content returned to the control level, whereas guanylate cyclase activity still increased over the level measured after the first treatment. It appears that insulin imprinting altered the calmodulin dependent guanylate cyclase regulation in Tetrahymena, and caused a switch-over to an 'energy-saving' system through decelerating the breakdown of cGMP by phosphodiesterase. PMID- 2572952 TI - The partial opiate receptor agonists, dezocine and ciramadol act as mu receptor antagonists at the feline ileocecal sphincter. AB - The effects of two novel synthetic narcotic agonist/antagonists dezocine and ciramadol were examined at the ileocecal sphincter (ICS) in the intact anesthetized cat. Changes in blood pressure were seen with higher doses of both dezocine and ciramadol. No ICS pressure changes were seen in the ICS to dezocine and an increase in ICS pressure was seen only to the highest dose of ciramadol examined (10 mg/kg). The antagonist action of the two drugs were examined against submaximal doses of the mu receptor agonist morphine sulfate, delta receptor agonist methionine enkephalin and the k-receptor agonist dynorphin. Both drugs inhibit the ICS response to morphine sulfate. No inhibition of the responses to methionine enkephalin or dynorphin were seen with dezocine and only partial inhibition of the ICS response to dynorphin was seen with ciramadol. PMID- 2572955 TI - Women's health in the 1990s. PMID- 2572954 TI - Ultrasound in medicine and biology. PMID- 2572956 TI - The acquired immunodeficiency syndrome and mosquitoes. PMID- 2572957 TI - Patient-controlled analgesia. PMID- 2572958 TI - [Bloodsucking mosquitoes (Culicidae, Diptera) of the Kindia area (the Republic of Guinea). 2. The genera Eretmapodites, Mansonia, Mimomyia, Malaya, Uranotaenia and Toxorhynchites]. AB - Results of the 2-year studies of fauna and habitats of the bloodsucking mosquitoes Eretmapodites, Mansonia, Mimomyia, Malaya, Uranotaenia, Toxorhynchites performed in the southwest of the Republic of Guinea in 1982-1983 are presented. 1222 mosquito specimen were identified, which belonged to 41 species and 6 aforementioned genus. 29 species and 1 genus (Malaya) were found in Guinea for the first time. Mosquito populations of the gallery forest, mango and humid savanna forests, marshes and habitations were studied. PMID- 2572959 TI - Epidural and subarachnoid opiates in obstetrics. PMID- 2572960 TI - Identification of the multidrug resistance-related P-glycoprotein as a cyclosporine binding protein. AB - The immunosuppressive agent cyclosporine A has been shown to reverse multidrug resistance (MDR) in malignant cells. In the present study, a 3H-cyclosporine diazirine analogue was used to photolabel viable MDR Chinese hamster ovary cells. The 170-kDa membrane P-glycoprotein, which functions as a drug efflux pump, was strongly labeled. The binding of 3H-cyclosporine diazirine analogue to P glycoprotein was competable by excess cyclosporine A and by the nonimmunosuppressive cyclosporine H. These results suggest that cyclosporine reverses the MDR phenotype by binding directly to P-glycoprotein and that this binding is not dependent on the immunosuppressive potential of the cyclosporine derivative. The identification of P-glycoprotein as a cyclosporine binding protein has obvious implications for cancer chemotherapy. PMID- 2572961 TI - Mercuric ions are potent noncompetitive antagonists of human brain kainate receptors expressed in Xenopus oocytes. AB - Kainate receptors are one of the major subtypes of excitatory amino acid receptors in the vertebrate central nervous system. Using Xenopus oocytes injected with RNA from human temporal cortex, it is possible to detect electrophysiologically the expression of this receptor subtype in these cells. Ions of the group IIb elements, particularly mercuric ions, are highly potent, noncompetitive inhibitors of these human brain kainate receptors. Mercury containing sulfhydryl reagents are also very effective, irreversible blockers of the kainate-gated currents of these oocytes. The recovery of kainate-activated currents after washout of Hg2+ is slow and incomplete relative to that seen after treatment either with Cd2+ or Zn2+. Cysteine or dithiothreitol can accelerate this recovery of kainate-inducible currents after Hg2+ inhibition. Besides the toxicological implications of these results, mercury compounds may be useful for future studies of the structure and physiology of the kainate receptor-channel complex. PMID- 2572962 TI - Dynorphin-selective inhibition of adenylyl cyclase in guinea pig cerebellum membranes. AB - Guinea pig cerebellum, which contains kappa opioid receptors uncontaminated by other opioid receptor types, was chosen to examine whether kappa receptors are coupled to adenylyl cyclase. Membranes were prepared from guinea pig cerebellum and pretreated at pH 4.5 to increase inhibitory activity, and adenylyl cyclase was assayed in the presence of dynorphin analogs as prototypical kappa agonists. Results showed that several dynorphin analogs inhibited adenylyl cyclase by 30 50%, whereas mu- and delta-preferring agonists had no effect. Dynorphin A and the kappa-selective compounds D-Pro10-dynorphin(1-11) and U-50,488H were the most potent agonists, with IC50 values of 0.03-0.05 microM, whereas other dynorphin gene products like dynorphin B and alpha-neo-endorphin were approximately 10-fold less potent. Like other Gi-coupled responses, dynorphin-inhibited adenylyl cyclase required GTP and sodium. Naloxone was a competitive antagonist for dynorphin-inhibited adenylyl cyclase, with 1 microM naloxone shifting the IC50 value of dynorphin A by 20-fold. The kappa-selective antagonist nor binaltorphimine was even more potent, with 0.1 microM nor-binaltorphimine shifting the dynorphin IC50 value by 50-fold. These results suggest that dynorphin A and its analogs inhibit adenylyl cyclase by binding to a guanine nucleotide-binding protein-coupled opioid receptor whose pharmacological specificity matches those of kappa receptors. PMID- 2572963 TI - Polymorphism of three HLA-DR7 bearing major histocompatibility complex extended haplotypes. AB - We have studied the complexity of HLA class II region in DR7 bearing extended haplotypes by restriction fragment length polymorphism (RFLP). Genomic DNA from homozygous cell lines and from unrelated individuals was digested with a number of restriction endonucleases and probed with DR alpha, DR beta, DQ alpha and DQ beta cDNA probes. We detected RFLPs that distinguished subspecificities of DRA, DRB1, DQA1 and DQB1 chain genes. On the basis of polymorphism in these genes, three distinct types of DR7 bearing extended haplotypes could be identified: (1) B44 or Bw47 or B14, DR7a (DRA1, B1.1), DRw53a (DRA1,B4), DQw2 (DQA1.1, B1.1); (2) B13 or B40, DR7b (DRA1, B1.2), DRw53a (DRA1, B4), DQw2 (DQA1.1, B1.1); and (3) Bw57, DR7c (DRA2, B1.2), DRw53b (DRA2, B4), DQw9 (DQA1.2, B1.2). Available evidence indicates that independent examples belonging to a haplotype were similar in RFLP patterns, suggesting that most examples of an extended haplotype belonging to a subtype are similar. The results in the present study have important implications for immune function and disease susceptibility. PMID- 2572964 TI - Betel nut-induced extrapyramidal syndrome: an unusual drug interaction. AB - Two cases are described of chronic schizophrenic patients maintained on depot neuroleptics, who developed severe extrapyramidal symptoms following a period of heavy betel nut consumption. A mechanism for this effect is proposed based on the pharmacological antagonism of the anticholinergic agent, procyclidine, by the active alkaloid ingredient of the betel, arecoline. PMID- 2572965 TI - Modulation of Trypanosoma cruzi adhesion to host muscle cell membranes by ligands of muscarinic cholinergic and beta adrenergic receptors. AB - Plasma membrane vesicles (PMVs) of Trypanosoma cruzi adhered to L6 myoblast host cells as a function of time and concentration in saturation phenomena in a similar fashion to that reported in a previous publication. The initial adhesion rate (A0) of T. cruzi PMVs to L6 myoblasts in tissue culture was inhibited by acetylcholine (10(-5) M), isoproterenol (10(-5) M) and norepinephrine (10(-8) M) (range 29.1-50.3% of control). Atropine, the antagonist of muscarinic cholinergic receptors (10(-5) M), and propranolol or pindolol, the antagonists of beta adrenergic receptors (10(-5) M), were also equal inhibitors of the T. cruzi PMV to L6 myoblast adhesion rate (range 26.1 to 55.5% of control). The alpha adrenergic receptor ligands yohimbine and phentolamine (10(-5) M) showed no A0 inhibitory activity in similar assays. The interaction of T. cruzi PMVs with type I host muscle sarcolemma receptors was clearly defined in assays which used porcine heart atrial membranes (PAMs) immobilized on cationic polyacrylamide beads. In this parasite membrane-host cell membrane assay system, 10(-10) M atropine and 5 x 10(-9) M propranolol produced a shift of an S-shaped T. cruzi PMV to PAM saturation isotherm to the right, suggesting that a negative cooperative interaction was produced between the intramembrane ligand binding site and another, surface heterotropic T. cruzi PMV adhesion site. Atropine and propranolol were equieffective inhibitors of the T. cruzi striated muscle sarcolemma recognition process, raising the possibility that T. cruzi attachment molecules annexed pairs of muscarinic cholinergic and beta-adrenergic receptors to effect adhesion of the two membrane surfaces. PMID- 2572966 TI - A new approach to the treatment of asthma. AB - Asthma is a chronic inflammatory condition. The previous emphasis on bronchodilator therapy, which does not treat the underlying inflammation, may be misplaced. Earlier introduction of antiinflammatory agents, such as corticosteroids or cromolyn sodium, is strongly recommended. Effective suppression of airway inflammation reduces the need for bronchodilator therapy and may reduce the morbidity and, perhaps, mortality of asthma. PMID- 2572967 TI - DNA-binding domain ancestry. PMID- 2572968 TI - Complex lymphoid and epithelial thymic tumours in Thy1-myc transgenic mice. AB - T-lymphocyte development takes place mainly in the thymus, where stromal cells of epithelial and haemopoietic origin are involved in inductive and selective mechanisms, which enable specific lymphocyte populations to migrate to the periphery and establish a network of immune responses. Experiments with intact animals have clarified the precursor-product relationships between thymocyte subpopulations, but the molecular mechanisms of cell interactions in the thymus are difficult to study in vivo. In an attempt to expand thymic cell populations in vivo and maintain them in vitro for such studies, we directed high levels of expression of the murine c-myc proto-oncogene in transgenic mice by inserting it into the mouse Thy-1 transcriptional unit. Such mice develop thymic tumours which contain proliferating thymocytes and, interestingly, expanded populations of epithelial cells. Both cell types can be maintained in vitro. PMID- 2572969 TI - Protein structure. Chaperones, paperones. PMID- 2572970 TI - Chemical synthesis. Scaling molecular Everests. PMID- 2572972 TI - The putatively antipsychotic agent amperozide produces behavioural stimulation in the rat. A behavioural and biochemical characterization. AB - Amperozide (FG 5606; N-ethyl-4-[4',4'-bis(p-fluorophenyl)butyl]-1 piperazinecarboximide ) is a new putatively antipsychotic compound with a postulated 5-HT2 antagonistic profile. Somewhat surprisingly amperozide dose dependently induced a behavioural stimulation in reserpinized and in nonpretreated rats. The behaviour consisted of both forward and backward locomotion as well as forepaw circling and a grooming like behaviour. Since the behavioural pattern clearly differ from that produced by classical dopaminergic or serotonergic agonists (e.g. apomorphine or 8-hydroxy-2-(di-n propylamino)tetralin, 8-OH-DPAT), and has not been previously reported, we decided to investigate the origin of this effect. In the behavioural paradigms it was not possible to antagonize the amperozide stimulation in reserpinized rats with the dopamine receptor blockers haloperidol, raclopride or R(+)-7-chloro-8 hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1 H-3-benzazepine, SCH 23390. Neither the 5-HT2 receptor blocking agent ritanserin nor the tryptophan and tyrosine hydroxylase inhibitor DL-3,4-dihydroxy-phenyl-alpha-propylacetamide, H22/54, could block the motoric stimulation or the forepaw circling behaviour produced by amperozide. However, the noradrenaline synthesis inhibitor bis-(4 methyl-1-homopiperazinylthiocarbonyl)-disulfide, FLA 63, as well as the alpha adrenoceptor antagonist phenoxybenzamine, could partly inhibit the locomotor stimulation. Hence, noradrenaline seems to be, at least in part, involved in the behavioural stimulatory effect of amperozide. Biochemically amperozide had no effect on the dopamine synthesis rate (DOPA formation) in normal or reserpinized animals in the striatal or the limbic brain regions. In reserpinized animals amperozide also failed to antagonize the decrease in DOPA formation after apomorphine and 3-hydroxy-benzylhydrazine HCl, NSD 1015, in these regions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572971 TI - Presynaptic dopamine DA2-receptors in rabbit jejunal arteries. An electrophysiological study. AB - Excitatory junction potentials (e.j.ps) evoked by nerve stimulation with 15 pulses at 1 Hz were recorded from muscle cells of rabbit isolated jejunal arteries. LY 171555 1 mumol/l, SKF 38393 10 mumol/l, dopamine 10 mumol/l and clonidine 0.1 mumol/l depressed all e.j.ps in the train. The percentage inhibition was inversely related to the number of pulses. S- and R-sulpiride, 10 mumol/l, domperidone 1 mumol/l, SCH 23390 1 mumol/l and rauwolscine 1 mumol/l did not change, or even depressed the first e.j.ps. Of these compounds only S- and R sulpiride, 10 mumol/l and rauwolscine 1 mumol/l facilitated the late e.j.ps. The percentage facilitation increased with the number of pulses until a maximum was reached; rauwolscine 1 mumol/l had the largest effect. S- and R-sulpiride, 10 mumol/l, as well as domperidone 1 mumol/l antagonized the action of LY 171555 1 mumol/l. S-Sulpiride was more potent than its R-isomer. SCH 23390 1 mumol/l and rauwolscine 1 mumol/l blunted the effect of SKF 38393 10 mumol/l. Rauwolscine 1 mumol/l slightly reduced the inhibition by dopamine 10 mumol/l; S-sulpiride 10 mumol/l was antagonistic only in the presence of rauwolscine 1 mumol/l. When rauwolscine 1 mumol/l, prazosin 0.1 mumol/l, propranolol 1 mumol/l and cocaine 10 mumol/l was added to the medium, dopamine 10 mumol/l continued to produce the same depression of e.j.ps, as in the absence of these compounds. Under such conditions S-sulpiride 10 mumol/l also counteracted dopamine 10 mumol/l. Rauwolscine 1 mumol/l prevented the effect of clonidine 0.1 mumol/l. The antagonists were not absolutely selective against only one type of agonist. We suggest that both presynaptic DA2- and postsynaptic DA1-receptors are present in rabbit jejunal arteries. The activation of either receptor-type may depress the e.j.ps. Dopamine interferes with neuroeffector transmission due to alpha 2 adrenoceptor agonist properties; its DA2-effect is unmasked only after alpha 2 adrenoceptor blockade. There was no evidence for a co-transmitter function of dopamine. PMID- 2572973 TI - Mode of action of tiaspirone on the central cholinergic system. AB - Tiaspirone, a potential antipsychotic drug, reduced the acetylcholine content of rat hemispheric brain regions (striatum 35%, hippocampus 20%, cortex 32% with no effect on N. accumbens) at an oral dose of 40 mg/kg. Choline content was uniformly raised in the same brain regions. A kinetic study showed that the drug is evenly distributed in the brain. Tiaspirone's effects on acetylcholine and choline in the striatum were not related in time. The fall off (30-240 min) of tiaspirone's effect on choline content paralleled the decline in striatal drug concentration (t1/2 = 240 min) whereas that on acetylcholine did not. No tolerance was observed to an acute challenge with tiaspirone on acetylcholine and choline in the striatum after 11 days' subchronic treatment. In vitro the drug had no effect on striatal choline acetyltransferase and acetylcholinesterase activities up to a concentration of 300 microM. The muscarinic agonist oxotremorine did not interfere with the acetylcholine decrease produced by the drug suggesting that muscarinic receptors are not essential for this effect. Tiaspirone, however, was found to be a competitive, reversible inhibitor of the sodium-dependent high-affinity choline uptake (SDHACU) by crude hippocampal and striatal synaptosomal preparations, giving IC50 values of respectively 3.69 microM and 1.14 microM. The compound did not alter SDHACU ex vivo despite the fact that it readily crosses the blood-brain barrier and achieves brain concentrations equivalent to its in vitro IC50 concentration. Tiaspirone antagonized the striatal acetylcholine increasing effect of apomorphine, a selective dopaminergic receptor agonist, supporting the idea that the drug affects the striatal cholinergic system by a primary action on dopamine receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572974 TI - Inhibition of 3H-noradrenaline accumulation by dopexamine hydrochloride in the isolated aorta of the rabbit. AB - The aim of the present work was to study the ability of dopexamine hydrochloride to interfere with the neuronal and extraneuronal uptake mechanisms by investigating the effect of dopexamine hydrochloride on 3H-noradrenaline accumulation by rabbit isolated aorta. Dopexamine hydrochloride (3 x 10(-9) - 10( 5) mol/l) reduced the accumulation of tritium by aorta incubated with 3H noradrenaline (10(-8) mol/l). The effect of dopexamine was compared to cocaine, dopamine, dobutamine, ADTN [(+/-)-2-amino-6,7-dihydroxy-1,2,3,4 tetrahydronaphthalene], ouabain and isoprenaline. Dopexamine hydrochloride (3 x 10(-9)-10(-7) mol/l) caused the same degree of inhibition irrespective of whether corticosterone (4 x 10(-5) mol/l) was present or not. The order of inhibitory potency was: desipramine greater than dopexamine hydrochloride greater than dopamine greater than ADTN greater than or equal to cocaine greater than dobutamine greater than ouabain greater than isoprenaline. In the presence of desipramine (10(-6) mol/l), corticosterone (10(-6)-10(-4) mol/l), but not dopexamine hydrochloride (10(-6) - 10(-4) mol/l), reduced the 3H-accumulation. It is concluded that dopexamine hydrochloride is a potent inhibitor of uptake-1 in rabbit isolated aorta. Dopexamine hydrochloride has no affinity for the uptake-2 mechanism in this tissue. PMID- 2572975 TI - 5-Hydroxytryptamine 5-HT1B and 5-HT1D receptors mediating inhibition of adenylate cyclase activity. Pharmacological comparison with special reference to the effects of yohimbine, rauwolscine and some beta-adrenoceptor antagonists. AB - 5-Hydroxytryptamine1B (5-HT1B) receptor mediated-inhibition of forskolin stimulated adenylate cyclase activity in rat substantia nigra was characterized pharmacologically and compared to 5-HT1D receptor mediated-inhibition of forskolin-stimulated adenylate cyclase activity in calf substantia nigra. Special attention was paid to the effects of drugs known to bind with high affinity to 5 HT1B (pindolol, propranolol, cyanopindolol, SDZ 21-009, isamoltane) or 5-HT1D recognition sites (yohimbine, rauwolscine). pEC50 or pKB values of a variety of 5 HT-receptor ligands (6 agonists including 5-HT, and 12 antagonists) for the inhibition of adenylate cyclase activity in rat substantia nigra, correlated significantly to the corresponding pKD values at 5-HT1B binding sites (r = 0.90, P = 0.0001). Amongst the alpha 2- and beta-adrenoceptor antagonists tested, none of the drugs expressed more than 35% of the intrinsic activity of 5-HT at 5-HT1B receptors. When tested as antagonists, their pKB values were in good agreement with their pKD values for 5-HT1B sites. By contrast, these drugs displayed marked intrinsic activity at 5-HT1D receptors: their pEC50 values were close to their pKD values for 5-HT1D sites and their effects could be potently antagonized by methiothepin. The rank orders of potency of the tested compounds at 5-HT1B and 5 HT1D were markedly different. The results strengthen the identity between 5-HT receptors mediating inhibition of adenylate cyclase activity in rat and calf substantia nigra and 5-HT1B and 5-HT1D binding sites, respectively. They underline the differences between these receptors in terms of intrinsic activities and potencies of drugs. PMID- 2572977 TI - [Pharmacotherapy in light and intermediate hypertension]. PMID- 2572976 TI - An insulin-releasing property of imidazoline derivatives is not limited to compounds that block alpha-adrenoceptors. AB - As we have demonstrated previously phentolamine stimulates the release of additional insulin from isolated mouse islets and raises plasma insulin levels in the whole rat. This effect was independent of the well known property of phentolamine to block alpha-adrenoceptors. In experiments on isolated pancreatic islets from mice we now demonstrate that tolazoline and antazoline which are chemically closely related to phentolamine, share its ability to potentiate insulin release. The following results were taken as evidence that this effect does not result from an alpha-adrenoceptor blocking action of imidazoline compounds. More than 10 times higher concentrations of phentolamine were required to liberate additional insulin from isolated islets than were effective in counteracting the inhibitory effect of clonidine on insulin release. The newly introduced alpha 2-adrenoceptor antagonist BDF 8933, which is an imidazoline derivative, stimulates insulin release as well, while the irreversible alpha adrenoceptor blocking agent benextramine of different structure failed to do so, even when being present in concentrations blocking the alpha 2-adrenoceptor mediated effects of clonidine. Antazoline shared the ability of phentolamine to stimulate insulin release despite having no or only very little alpha adrenoceptor blocking activity. When used under our conditions, it almost entirely failed to alleviate the inhibition of insulin release induced by clonidine. We conclude that the response of the islet cells to imidazoline derivatives is not limited to those capable of blocking alpha-adrenoceptors. On the other hand, alpha-adrenoceptor blocking agents of different chemical structure fail to induce the release of additional insulin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572978 TI - [Another case of Hantaan virus infection in The Netherlands]. AB - In a serological survey among Dutch patients suspected of leptospirosis, using a recently developed enzyme-linked immunosorbent assay, a patient was traced with a high antibody titre to Hantaan virus. No anti-leptospira antibodies were detected in this 27-year-old man. Shortly before he had been admitted to the hospital with progressive dyspnoea and coughing, accompanied with high fever. An interstitial pneumonia was diagnosed. He subsequently developed a progressive renal failure with proteinuria and polyuria. Later a liver failure accompanied with thrombocytopenia, anaemia and coagulation disturbances occurred. Before an aetiological diagnosis was made, the patient was treated with erythromycin. The patient eventually recovered completely. Based on the clinical symptoms and the positive serology, it was concluded that the disease diagnosed had probably been caused by a Hantaan virus infection. The diagnostic value of Hantaan virus serology in patients with similar symptoms is stressed. PMID- 2572979 TI - [Significance of the registration of testis localization in child health services for the avoidance of unnecessary orchiopexies]. AB - The health records of 1816 11-year-old Dutch schoolboys were examined to assess the importance of these records for boys with an apparently non-scrotal testicle. Full records from birth onward existed for 58% of the boys; at least 4 examinations were recorded in 76% and no prior examinations in less than 4% of the boys. Orchidopexy had been performed in 2.9% of the boys; the records of 33 of the 53 boys with a history of orchidopexy showed intrascrotal position of both testicles at least once. The conclusion is that the Dutch youth health system possesses relevant records which should be consulted by practising physicians and surgeons before orchidopexy is considered. PMID- 2572980 TI - [The role of the adenyl cyclase system in cholinergic modulation of synaptic transmission in the hippocampus]. AB - The adenylate cyclase system has been studied from the standpoint of its significance in cholinergic modulation of the synaptic transmission in the CA1 field of the rat hippocampal slices. Microionphoretic application of ACh as well as addition of either carbachol or tolbutamide (an inhibitor of cAMP-dependent protein kinase) blocked the transmission in synapses formed by the Schaffer collaterals and commissural fibres with dendrites of carbacholine both the number of releasing quanta of the neurotransmitter and the probability of their release decreased. Atropine eliminated the inhibitory effect of carbacholine on synaptic transmission. Dibutyryl cAMP and forskolin increased the amplitude of synaptic potentials and completely or partially prevented the inhibitory effect of cholinomimetics on synaptic potentials. The results obtained revealed opposite effects of cholinomimetics and activators of the adenylate cyclase system on neurotransmission in synapses formed by the Schaffer collaterals/commissural fibres and dendrites of pyramidal neurons of the hippocampal CA1 field. PMID- 2572982 TI - [A statistical analysis of the change in spontaneous quantum secretion of a mediator in the presence of carbacholine in rat neuromuscular synapse]. AB - Time of appearance and amplitudes of miniature end-plate potentials were studied statistically during the carbacholine action on the rat soleus muscle. Facilitating effect of carbacholine on the spontaneous quantal transmitter release did not change time distribution of miniature end-plate potentials. It is suggested that this effect of carbacholine was rather due to an increase in the activity of releasing sites, than due to an increase in their number. PMID- 2572981 TI - [The effect of carbacholine on spontaneous quantum secretion of a mediator from frog motor nerve endings in the presence of ouabain and in a potassium-free medium]. AB - The carbacholine (Cch) effect on the frequency of miniature end-plate potentials (MEPP) has been studied in the frog muscle in the presence of ouabain and in the potassium-free solution. The depressing presynaptic effect of Cch on the MEPP frequency was eliminated by ouabain, whereas the inactivation of ATPase by the potassium-free solution did not affect the degree of the MEPP frequency decrease. The possibility of the spontaneous quantal release regulation in cholinomimetic drugs by means of the ouabain-sensitive mechanism not related to the pumping function of the Na+, K+ and ATPase active transport is discussed. PMID- 2572983 TI - N-methyl-D-aspartate receptors and Alzheimer's disease. AB - The results of several studies now suggest that the density of N-methyl-D aspartate (NMDA) receptors is maintained in many Alzheimer's disease (AD) cases, although loss of these receptors can occur in specific regions as a consequence of severe neuronal loss. Recent findings demonstrate that there are at least two subtypes of the NMDA receptor which are allosterically regulated. To determine the status of the NMDA receptor in AD, studies are required which will examine the activation state of the NMDA receptor and the properties of subtypes in relation to neuronal density and structure. PMID- 2572984 TI - Selectivity of excitotoxic mechanisms in Alzheimer's disease. AB - Dysfunction of glutamatergic neurons in Alzheimer's disease may actually participate in some of the neurodegenerative processes via potential excitotoxic mechanisms. Two hypotheses are advanced which describe how a generalized excitotoxic mechanism could interact with unique metabolic properties of specific cell types leading to their selective vulnerability in Alzheimer's disease. PMID- 2572986 TI - Novel CNS-directed drug delivery systems in Alzheimer's disease and other neurological disorders. AB - Increasing knowledge of the neurochemical aspects of central nervous system function raises the possibility of treating Alzheimer's disease (AD) and other neurological diseases by the appropriate manipulation of neurotransmitters, neuromodulators, neurohormones or neurotrophic factors. Clinical application of this knowledge may, however, be inhibited by long standing problems with drug delivery to the central nervous system (CNS). Novel, CNS-directed, drug delivery systems might be used to overcome many of these problems. The problems encountered in drug delivery to the brain, present experience with the clinical use of some novel drug delivery systems and the advantages and disadvantages of these systems will be discussed. PMID- 2572987 TI - The necessity of human trials of putative therapies for Alzheimer's disease. AB - The absence of a suitable animal model for Alzheimer's disease leaves therapeutic trials in human subjects as a necessity. Reasonable criteria can be formulated for deciding which therapies should be tested. Scientific rationale and likelihood of success should be major considerations. Novel approaches of delivering drugs to the central nervous system should not be discouraged so long as complication rates can be shown to be low. PMID- 2572988 TI - Methodological considerations in cognitive disorders or: "the baby needs enough bath water". AB - Establishing treatment efficacy for neuropsychiatric disorders is an often protracted process due to vagaries of symptoms, course, and probably the inherent variability of the central nervous system. No single study design is likely to be definitive. Thorough evaluation of intracranial drug infusion for illnesses such as Alzheimer's Disease will require extended, multidisciplinary work before true evaluation can confidently be made. PMID- 2572985 TI - The role of neuronal energy in the neurotoxicity of excitatory amino acids. AB - Excitatory amino acids, acting at receptors such as the N-methyl-D-aspartate (NMDA) subtype, are good candidates for a major role in the neuronal death characteristic of Alzheimer's disease. Recent evidence from studies with cultured neurons suggests that perturbations in the energy metabolism of the neuron may be involved in the transition of NMDA agonists from neurotransmitters to neurotoxins via a mechanism that involves relief of the voltage-dependent Mg++ block of the NMDA channel. PMID- 2572989 TI - [Primary interhemispheric subdural abscess: report of a case]. AB - We reported a rare case of primary interhemispheric subdural abscess. Twenty three cases of this pathological condition have been reviewed. In those reports, however, findings of magnetic resonance imaging (MRI) were not referred to. In this report, MRI findings in this pathological condition, in addition to X-ray computed tomography (CT), is mentioned. A 22-year-old man, who had suffered from headache and vomiting for 2 weeks, suddenly became drowsy and left-hemiparetic. The X-ray CT scan on admission showed a well-circumscribed low density area in contact with the falx in the right parieto-occipital region. This lesion had so called "ring enhancement". MRI in the sagittal view revealed that, along the falx, the long T1 and T2 areas extended from the right cerebellar tentorium to the right frontal region. The operation demonstrated the capsular formation of the abscess. After pus aspiration, continuous drainage was performed from the cavity of the abscess. The patient fully recovered postoperatively. In the diagnosis of interhemispheric subdural abscess, it is said that conventional X ray CT sufficiently reveals the quality of the lesion, the precise site, and the anatomical relation to the surrounding edema. In our case, MRI was able to confirm the diagnosis made by the X-ray CT. Furthermore, MRI was thought to be superior to the X-ray CT in the evaluation of the extension of the abscess and in the delineation of the surrounding edema. Combined use of X-ray CT and MRI in cases of interhemispheric abscess was considered to make the diagnosis more precise in both qualitative and quantitative aspects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2572990 TI - Beta-endorphin mediates clonidine stimulated growth hormone release. AB - The role of beta-endorphin in the stimulation of growth hormone secretion elicited by administration of clonidine (Clon), an alpha2-adrenergic agonist, was investigated in awake, freely moving male rats. Animals were infused slowly with either 1 ml of normal rabbit serum (NRS), beta-endorphin antiserum (beta-end-AS) or ACTH antiserum (ACTH-AS) 2 h before the administration of Clon (100 microgram/kg body weight, intravenously). In addition, naloxone (Nal) (2.5 mg/kg body weight, intravenously) was given 15 min prior to Clon in some experiments. Blood samples were taken at 15-min intervals prior to and following Clon administration. Clon caused plasma GH levels to rise 15-fold to peak levels of 177 +/- 38 ng/ml (p less than 0.01) at 30 min. Pretreatment of both Nal or beta end-AS significantly reduced Clon-stimulated GH secretion to 72 +/- 19 ng/ml (p less than 0.05) and 87 +/- 30 ng/ml (p less than 0.05) respectively. In contrast, the infusion of ACTH antiserum did not affect Clon-stimulated GH release. Our data suggest that beta-endorphin or a related opioid peptide is an important mediator of GH secretion induced by alpha2-adrenergic stimulation. Since blockade of opioid receptors blunted Clon-induced GH release only partially (approximately 50%), other mediators are most likely activated following alpha 2-adrenergic stimulation. PMID- 2572991 TI - Involvement of extracellular calcium and arachidonate in [3H] dopamine release from rat tuberoinfundibular neurons. AB - The mechanism of [3H]dopamine [( 3H]DA) release was investigated using primary cultures of dispersed cells from the rat tuberoinfundibular region, which contains tyrosine hydroxylase (TH)-like immunoreactive neurons. The calcium ionophore A23187 at 10 nM and above caused a significant and dose-dependent increase in [3H]DA release. In the presence of 50 microM A23187, [3H]DA release was detectable within 30 s and reached a plateau in 15 min. The induction of [3H]DA release by 50 microM A23187 was abolished by lowering the extracellular calcium concentration with 2 mM EDTA. Maitotoxin, another calcium-channel activator, also increased [3H]DA release at a concentration of 50 ng/ml. Exogenous additions of 100 mIU/ml phospholipase A2 and 10 microM arachidonate caused significant release of [3H]DA. Furthermore, A23187 stimulated [3H]arachidonate release from tuberoinfundibular dopaminergic (TIDA) neurons in a dose- and time-dependent manner. These results suggest that extracellular calcium and arachidonate are involved in the process of [3H]DA release from rat TIDA neurons. PMID- 2572993 TI - Penicillin-induced bursting in motoneurones of the frog spinal cord. Elimination by NMDA antagonist. AB - The effects of penicillin were investigated in lumbar motoneurones of isolated spinal cord preparation of the frog (Rana ridibunda). Spinal root discharges were recorded and single cell activity was studied with intracellular electrodes. Bath application of 500 IU/ml of penicillin G induced in motoneurones prolonged depolarization shifts, followed by repeated transient depolarizations lasting several hours. In all motoneurones studied, this bursting activity was synchronized with discharges recorded from the ventral root. Blockade of N-methyl D-aspartate (NMDA) receptors by D,L-2-amino-5-phosphonovaleric acid (50-100 microM) completely eliminated the bursting activity. It is suggested that the spinal cord may be an important locus of the anticonvulsant effect of NMDA antagonists. PMID- 2572992 TI - Characterisation of kappa-opioid binding sites in rat and guinea-pig spinal cord. AB - The binding of radiolabelled ligands with high affinity for kappa-opioid binding sites has been studied in homogenates of lumbo-sacral spinal cord from the rat. The unselective ligands [3H]bremazocine and [3H]diprenorphine labelled a large number of sites which could not be fully resolved in terms of mu-, delta- and kappa-types by displacement assays. In particular binding at the kappa-site appeared anomalous in that sites which could be identified as high affinity kappa type represented only 40% of total kappa-binding, defined using the unselective [3H]ligands. This was confirmed by the low levels of binding seen with the kappa agonists [3H]dynorphin A(1-9) and [3H]U-69593. In guinea-pig cord, under conditions in which binding to mu- and delta-sites was suppressed, [3H]dynorphin A(1-9) and [3H]U-69593 labelled only 60% of the kappa population, defined by the [3H]unselective ligands. The reasons for the observed discrepancies are discussed. PMID- 2572994 TI - Effects of methylenedioxymethamphetamine on local cerebral glucose utilization in the rat. AB - The effects of (+-)3,4-methylenedioxymethamphetamine (MDMA, "ecstasy") (5, 10, 15 or 30 mg/kg, i.p.) on local cerebral utilization of glucose were studied by the quantitative autoradiographic 2-deoxy-D-[1-14C]glucose method in awake adult male Fischer-344 rats. Statistically significant effects on local utilization of glucose, 5 min after the administration of MDMA were observed in 20 of 60 areas of brain sampled. Marked stimulation was seen in components of the extrapyramidal motor system (substantia nigra, globus pallidus, entopenduncular nucleus, subthalamic nucleus, cerebellar vermis). The limbic system showed decrements in the medial cortex and hippocampal dentate gyrus (outer blade) and the lateral habenula, while there was stimulation in the mammillary body and the basolateral amygdaloid nucleus. Glucose utilization in MDMA-treated rats was reduced in the superior colliculus and medial terminal nucleus of the accessory optic system, but was unchanged in the visual cortex and components of the auditory system. Some of the effects of MDMA on cerebral utilization of glucose resembled those previously reported with l-cocaine, d-amphetamine, and phencyclidine, implicating some common mechanisms in the actions of these drugs. PMID- 2572995 TI - Effects of anxiolytic and anxiogenic drugs on exploratory activity in a simple model of anxiety in mice. AB - Chlordizaepoxide, pentylenetrazole, phenobarbital, N-methyl-beta-carboline-3 carboxamide (FG-7142), buspirone and the novel serotonin3 receptor (5-HT3) antagonist, 3-tropanyl-indole-3-carboxylate (ICS 205-930), were examined in the two-compartment exploratory model (Crawley and Goodwin, 1980). The results indicated that, utilizing the time mice spend in the dark side of the apparatus as an index of anxiety, increased the sensitivity of the model and enabled both anxiolytic and anxiogenic agents to be detected. PMID- 2572996 TI - Excitatory amino acid receptor-mediated activation of solitarial deglutitive loci. AB - The deglutitive actions of glutamate were investigated in urethane-anaesthetised rats in order to determine whether different excitatory amino acid receptors mediate activation of pattern generator elements contained within the nucleus tractus solitarii. When applied by micropneumophoresis (0.01-10 pmol) from multibarrelled glass micropipettes (tip diameter 2-5 microns), the excitatory amino acid-receptor agonists, N-methyl-D, L-aspartate (NMA), N-methyl-D-aspartate (NMDA), quisqualate and kainate displayed a rank order of potency at glutamate responsive pharyngeal sites, in the subnuclei ventralis and intermedialis, where KA greater than NMA/NMDA greater than QA; however, the potency followed the order NMA/NMDA greater than KA greater than QA at oesophageal sites within the subnucleus centralis. The NMDA-receptor blockers, 2-amino-5-phosphonovaleric acid (APV) and 2-amino-7-phosphonoheptanoic acid (AP7), selectively and reversibly inhibited the glutamate-evoked oesophageal responses, but had no corresponding effect on rhythmic oesophageal responses elicited by muscarine. At loci in the nucleus tractus solitarius, where glutamate elicited a complete swallowing sequence, APV/AP7 spared the pharyngeal component but selectively blocked the oesophageal component. The nonselective glutamate-receptor antagonist, gamma-D glutamylglycine suppressed both pharyngeal and oesophageal responses elicited by glutamate. It is concluded that different types of excitatory amino acid receptors are associated with the deglutitive premotor subnuclei of the nucleus tractus solitarii; kainate receptors predominate within the subnuclei ventralis and intermedialis and NMDA receptors within the subnucleus centralis. Both kainate- and NMDA-mediated mechanisms can operate under physiological conditions. PMID- 2572998 TI - Opioid-like activity in the cerebrospinal fluid of pain patients treated by electroacupuncture. AB - Thirteen patients with pain from various causes were treated by electroacupuncture for 30 min. Cerebrospinal fluid (CSF) was obtained before and after treatment. Opioid-like substances in the CSF were fractionated by high pressure liquid chromatography and assayed by competitive receptor binding using a mu-specific radioligand, [D-ala2, MePhe4, gly-ol5]-enkephalin (DAGO). Opioid activity, associated with a fraction, eluted at 18-20% acetonitrile, consistently showed an increase in level after acupuncture. Two other fractions eluted at larger concentrations of acetonitrile also increased significantly after acupuncture; however the increase was not consistently observed in every patient. Measurements of beta-endorphin and dynorphin by radioimmunoassay indicated that 80 and 60% of the patients, respectively, had a higher level of these peptides after acupuncture. The nature of the opioid activity, eluted at 18-20% acetonitrile is unknown; however a small amount of it could be found in various parts of the brain of rat. PMID- 2572997 TI - Electrophysiological effects of neurotensin on dopaminergic neurones of the ventral tegmental area of the rat in vitro. AB - The effects of neurotensin on the spontaneous firing rate of presumed dopaminergic neurones of the ventral tegmental area of the rat, were studied in a slice preparation of brain by extracellular single-cell recordings. Bath-applied neurotensin excited all cells which were studied (N = 25). This effect was concentration-dependent; the threshold was 10(-10) M and maximal activation (about 30 spikes/10 sec) was obtained with 10(-6) M. The EC50 (half-maximal effective concentration) was roughly estimated at 35 nM. The action of neurotensin was mimicked by neurotensin 8-13 (N = 6), but not neurotensin 1-8 (N = 6). It persisted in low-calcium, high-magnesium solutions (N = 5) and therefore probably resulted from a direct activation of neurotensin receptors. The responses to neurotensin were long-lasting (30-60 min after a 10 min 10(-7) M infusion) and exhibited little tachyphylaxis. Dose-response curves to the dopaminergic agonist BHT920 showed that, during the infusion of 10(-7) M neurotensin, dopaminergic autoreceptors of some neurones were less sensitive than in control conditions. This was not a non-specific effect produced by the excitation, since it was not observed during the infusion of another excitant, N methyl-D-aspartate (NMDA). These results show that neurotensin potently activates presumed dopaminergic neurones in the ventral tegmental area in vitro; it may also decrease the effectiveness of the autoreceptors of some neurones. PMID- 2572999 TI - Distribution of neuropeptides in the infundibular nucleus of the sheep. AB - The distribution of neurons exhibiting somatostatin (SRIF)-, neuropeptide Y (NPY) , beta-endorphin- and neurotensin (NT)-like immunoreactivity within the infundibular nucleus (NI) of the sheep, and the extent of coexistence of the above peptides within individual neurons of the NI were investigated with immunocytochemical techniques. Our results show that the above neurochemical types of neurons exhibit specific and largely non-overlapping patterns of distribution within the NI of the sheep. Furthermore, the coexistence of these peptides within neurons of the NI is very limited, as from all possible permutations checked, only SRIF and NPY were found together in a small number of cells. PMID- 2573000 TI - Selectivity of various opioid peptides towards delta-, kappa; and mu-opioid receptors mediating presynaptic inhibition of neurotransmitter release in the brain. AB - The selectivity of a series of opioid peptides towards the mu-, delta- and kappa opioid receptors mediating differential inhibition of electrically-induced neurotransmitter release from rat brain slices was studied, viz. cortical [3H]noradrenaline release (inhibited via mu-receptors), striatal [3H]dopamine release (inhibited via kappa-receptors) and striatal [14C] acetylcholine release (inhibited via delta-receptors). The highest affinity pD2 7.4) and selectivity towards mu-receptors was exhibited by Tyr-D-Ala-Gly-(NMe)Phe-Gly-ol (DAGO), whereas [D-Pen2, D-Pen5]enkephalin (DPDPE) was found to be the most selective delta-receptor agonist (pD2 7.3). Also the hexapeptides [D-Ser2]Leu-enkephalin Thr (DSLET) and [D-Thr2]Leu-enkephalin-Thr (DTLET) showed a relatively high selectivity and, in addition, a high affinity (pD2 8.2-8.4) for delta-opioid receptors. Both dynorphin(1-13) and dynorphin(1-8) exhibited a high affinity for kappa-receptors (pD2 resp. 8.3 and 8.0), but the latter was far less selective. Both of the dynorphin A-related peptides showed affinity to mu-receptors (pD2 6.7 6.8), but dynorphin(1-8), in contrast to dynorphin(1-13), also displayed a high affinity to delta-receptors (pD2 7.6). PMID- 2573002 TI - An approach to the development of drugs for appetite disorders. AB - This review covers some modern concepts in the development of drugs to treat appetite disorders. Specific attention is paid to the peripheral satiety system and the role of gastrointestinal peptides such as cholecystokinin in the pathogenesis of satiety. Alterations in neuropeptide Y and/or peptide YY are suggested to play a role in the pathophysiology of bulimia. Corticotropin releasing factor is a putative candidate peptide involved in anorexia nervosa. The serotonin reuptake inhibitors fenfluramine and fluoxetene decrease weight in obese subjects. Endogenous opioids modulate the choice of palatable foods. Anorexia in the old appears to be related to a decrease in opioid feeding drive and an excess of the satiety action of cholecystokinin. Other agents involved in weight regulation include those which alter gastric emptying, increase thermogenesis, or modulate fat cell metabolism. It should be stressed that many neurotransmitters that modulate appetite also alter other behaviors, increasing their propensity to produce side effects. PMID- 2573001 TI - Drugs as research tools for elucidating behavioral processes. PMID- 2573003 TI - Is there a common dopaminergic basis of time perception and reaction time? AB - The effects of 3 mg haloperidol and 125 mg Madopar on duration discrimination (DD) as well as reaction times were tested in a placebo-controlled, double-blind crossover study with 24 healthy male volunteers. Performance in DD was significantly impaired under haloperidol compared to placebo as well as to Madopar. No changes could be demonstrated for Madopar compared to placebo. Similar results were obtained for measures of reaction time. Significant negative correlations between individual changes in DD and measures of reaction time under each drug condition revealed that subjects with drug-induced impairment in DD also show an increase in reaction times. The nearly identical patterns of drug induced changes in DD and measures of reaction time may be interpreted in terms of a common neural basis. PMID- 2573004 TI - Neurotransmitter diversity and its role in integrative brain functions. PMID- 2573005 TI - Tardive oculogyric crises. AB - Involuntary ocular deviations, or oculogyric crises (OGC), commonly occur in acute dystonic reactions to dopamine receptor blocking drugs (neuroleptics). We describe 4 patients with tardive OGC due to prolonged exposure to neuroleptics. In addition to the OGC, the patients had other tardive movement disorders. All patients improved with tetrabenazine. We conclude that tardive OGC are often not recognized and represent part of the spectrum of tardive dystonia. PMID- 2573007 TI - [Polyarteritis nodosa and pulmonary neoplasms. Report of a clinical case]. AB - A case of rare association between PAN and cancer is presented. The tumour was a bronchogenic carcinoma (large cell type) diagnosed about 14 years after the beginning of PAN, that in this case had a very long course increasingly affecting the principal arteries of the legs. PMID- 2573006 TI - Gerstmann-Straussler-Scheinker disease. II. Neurofibrillary tangles and plaques with PrP-amyloid coexist in an affected family. AB - Azzarelli et al reported an Indiana kindred affected by a hereditary disorder, characterized clinically by ataxia, parkinsonism, and dementia. Recently, we studied neuropathologically the 3rd and 4th cases that came to autopsy among the patients of this family. As in 2 patients examined previously, amyloid plaques were widespread throughout the cerebrum and the cerebellum, whereas neurofibrillary tangles were numerous in the cerebral cortex, the hippocampus, and the substantia innominata. Amyloid plaques were not recognized by polyclonal antibodies against the Alzheimer's disease amyloid A4 protein, but did contain epitopes recognized by antibodies against a prion protein. Spongiform changes were occasionally observed and were mild. Our findings indicate that this familial disorder is a form of or is related to Gerstmann-Straussler-Scheinker disease. The consistent presence of numerous neurofibrillary tangles may be important in differentiating a distinct subgroup of patients with familial Gerstmann-Straussler-Scheinker disease, and indicates that a disturbance of the cytoskeleton might be part of the neuronal pathology of Gerstmann-Straussler Scheinker disease. PMID- 2573008 TI - Topographic relations between tyrosine hydroxylase- and luteinizing hormone releasing hormone-immunoreactive fibers in the median eminence of adult rats. AB - Employing electron microscopic double immunolabeling, we determined a close apposition of tyrosine hydroxylase (TH) and luteinizing hormone-releasing hormone (LHRH) nerve fibers in the rat median eminence (ME). These axo-axonic contacts occurred frequently in the internal and palisade zones, i.e. at the level of the fiber preterminals. In the superficial area of the ME, major TH fibers abutted on the basal lamina and some were projected into the pericapillary space of the portal vessels. Conversely, LHRH fibers were arrested by the endfeet of tanycytes in reaching the basal lamina. PMID- 2573009 TI - Effects of histamine on thermosensitive neurons in rat preoptic slice preparations. AB - Single neuronal activities were recorded extracellularly from slice preparations of the rat preoptic area and effects of histamine (0.01 10 microM) on the activities were examined with regard to thermosensitivies of the neurons. Superfusion of histamine increased the firing rate in 52 of 75 warm-sensitive neurons and in 22 of 41 thermally insensitive neurons in a dose-dependent manner. Ten (3%) warm-sensitive neurons and 6 (15%) thermally insensitive neurons were inhibited by histamine. Mepyramine (10 microM) (H1-antagonist), but not famotidine (H2-antagonist), blocked the histamine (10 microM) induced excitation in 19 (76%) of 25 warm-sensitive neurons and in 6 (75%) of 8 thermally insensitive neurons. These results suggest that histamine excites both warm sensitive and thermally insensitive neurons in the preoptic area mainly via the H1-receptor. PMID- 2573011 TI - Reserpine does not prevent 3,4-methylenedioxymethamphetamine-induced neurotoxicity in the rat. AB - 3,4-Methylenedioxymethamphetamine (MDMA; 'Ecstasy') is a known neurotoxin to 5 hydroxytryptamine (5-HT) nerve terminals. Recent studies have suggested that endogenous dopamine (DA) and/or 5-HT may mediate the MDMA-induced neurotoxicity. The central monoamine stores of rats were significantly decreased with reserpine (5 mg/kg) prior to toxic injections of MDMA. Rats given MDMA (30 mg/kg) displayed significant decreases in the density of 5-HT nerve terminals labeled by [3H]paroxetine both with (51 +/- 8%) and without (43 +/- 20%) reserpine pre treatment. These data suggest that the degeneration of 5-HT nerve terminals following MDMA is independent of the presence of endogenous stores of DA or 5-HT. PMID- 2573010 TI - Differential effects of somatostatin on adenylate cyclase as functional correlate for different brain somatostatin receptor subpopulations. AB - In homogenates of rat hippocampus and striatum, but not substantia nigra, somatostatin (SRIF) inhibits forskolin-activated adenylate cyclase in nanomolar concentrations. However, SRIF can also stimulate adenylate cyclase in micromolar concentrations in homogenates of rat hippocampus and substantia nigra. The SRIF octapeptide SMS 201-995 solely inhibits the forskolin-activated adenylate cyclase in the 3 preparations. These results suggest that the SRIF-specific stimulation of adenylate cyclase may be a functional correlate for the brain-specific SRIF receptor subpopulation, whereas the SRIF and SMS 201-995 inhibition of stimulated adenylate cyclase correlate with the SRIF receptor subpopulation present in brain and non-neuronal tissues. PMID- 2573012 TI - Preganglionic nerve stimulation increases mRNA levels for tyrosine hydroxylase in the rat superior cervical ganglion. AB - Increased synaptic stimulation of sympathetic neurons in vivo causes a delayed increase in the activity and the amount of tyrosine hydroxylase (TH). To determine whether these changes result from an increase in the messenger RNA for TH, the rat preganglionic cervical sympathetic trunk was electrically stimulated unilaterally for 90 min, and 48 h later RNA was extracted from stimulated and contralateral control superior cervical ganglia. Northern blots probed with a cDNA for TH demonstrated that nerve stimulation produced about a 2.5-fold increase in the amount of TH mRNA in the ganglion. These results indicate that synaptic stimulation leads to an increase in TH mRNA, either by increasing the rate of transcription of the TH gene or by increasing the stability of its mRNA. PMID- 2573013 TI - Somatostatinergic neurones of the developing human and cat retinae. AB - We have examined somatostatin-immunoreactive (S-IR) neurones in developing retinae of the human and cat. At 14 and 16 weeks' gestation (G14 and G16) in the human, S-IR cells were only found close to the putative fovea centralis, but by 18 weeks' gestation (G18), they were located in all retinal regions. By adulthood, the majority of S-IR cells were restricted to inferior retina. In the developing cat retina, two classes of S-IR cells were recognized. S1-IR cells were similar in morphology and distribution to adult cells: they had small round somata which were only found in inferior retina and gave rise to beaded processes which traversed the inner plexiform layer (IPL) and nerve fibre layer (NFL). S2 IR cells had larger somata located in the ganglion cell layer (GCL) and the label was compartmentalized within their cytoplasm. Most S2-IR cells had lost immunoreactivity by P (postnatal day) 25 and may have been alpha-ganglion cells transiently expressing somatostatin in association with their retention of plasticity into postnatal life. PMID- 2573014 TI - The N-methyl-D-aspartate (NMDA) receptor complex: a stoichiometric analysis of radioligand binding domains. AB - A stoichiometric analysis of pharmacological domains within the N-methyl-D aspartate (NMDA) receptor complex was made by evaluating the binding of L [3H]glutamate, [3H]CPP, [3H]glycine and [3H]MK-801 to purified synaptic membranes isolated from rat telencephalon. The binding of all radioligands exhibited pharmacological and kinetic properties consistent with the labeling of homogeneous populations of sites associated with the NMDA receptor. However, strychnine-insensitive [3H]glycine binding sites were present at close to 2-fold the density of the other sites examined. These data, together with recent electrophysiological and receptor autoradiographic findings, are utilized as a basis for hypotheses regarding the ratio of transmitter recognition, allosteric and channel binding sites within the NMDA receptor complex. PMID- 2573015 TI - Effect of D1 receptor stimulation in normal and MPTP monkeys. AB - The effect of a selective agonist of the dopamine D1 receptor (SKF 38393) and of the D2 receptor (LY-171555) was tested acutely in normal and in monkeys with a parkinsonian syndrome induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The D2 agonist induced a strong locomotor response and lingual dyskinesia in both normal and parkinsonian monkeys. The D1 agonist however had no locomotor effect by itself but induced tongue protrusions in normal monkeys only. It appeared to potentiate the dyskinetic effect of LY 171555 in MPTP monkeys but it antagonized the locomotor action of the D2 agonist in both normal and MPTP monkeys. The selective D1 and D2 antagonists SCH 23390 and sulpiride were also tested. Both compounds were able to suppress the dyskinetic action of the combined agonists in normal animals but only the D2 antagonist was effective in the same conditions in MPTP monkeys. These findings emphasize the importance of the D2 receptor in mediating the locomotor response as well as dyskinesia in monkeys. PMID- 2573016 TI - Serotonin acts in the synaptic region of sensory neurons in Aplysia to enhance transmitter release. AB - An important mechanism that contributes to sensitization in Aplysia is heterosynaptic facilitation of the synaptic connections between sensory neurons (SNs) and motor neurons (MNs). Heterosynaptic facilitation, in turn, is associated with broadening of the spike in the SN. Spike broadening is readily observed in recordings from somata of SNs, and from growth cones of SNs in culture, but broadening in synaptic terminals has only been inferred. Intracellular recordings were made from somata of SNs and from somata of follower MNs. Additional recordings were made from the axons of SNs as they enter the neuropil in the pedal ganglion. Serotonin (5-HT) broadened action potentials in axons of SNs and enhanced excitatory postsynaptic potentials (EPSPs) in the MNs, even after the axons of SNs were surgically separated from their somata. These results indicate that both heterosynaptic facilitation and spike broadening in the axon are due to the local action of 5-HT and can occur independently of modulation of membrane properties in the soma. PMID- 2573017 TI - Spontaneous activity mediated by NMDA receptors in immature rat entorhinal cortex in vitro. AB - Intracellular recordings were made from cells in layer II of entorhinal cortex slices from 9 to 13-day-old rats. The majority of these cells showed pronounced spontaneous synaptic activity which could summate into large depolarizing events and give rise to bursts of spikes. These events could be associated with an apparent decrease in membrane conductance. They were reduced in amplitude by hyperpolarization of the cell and tended to increase on depolarization. Perfusion with 2-amino-5-phosphonovalerate abolished the spontaneous activity. Thus, in layer II of the entorhinal cortex at this stage of development there seems to be a functional enhancement of ongoing synaptic activity mediated via activation of N-methyl-D-aspartate receptors. PMID- 2573018 TI - Strain-dependent decrease in glutamate binding to the N-methyl-D-aspartic acid receptor during aging. AB - Glutamate binding to the N-methyl-D-aspartic acid (NMDA) receptor decreases in two strains of aged mice. In BALB/c mice the Bmax values decline 16% by 10 months and 45% by 30 months of age when compared to 3 months. The Kd increased more by 10 months (+29%) than by 30 months (+14%). In the C57Bl strain the Bmax was unaltered by 10 months but decreased 17% by 30 months. The Kd values, however, increased 121% by 10 months and 283% by 30 months of age. These data suggest that the age-related decline in glutamate binding to the NMDA receptor may predict a strain-specific reduction in NMDA-mediated processes (e.g. long-term potentiation, postsynaptic calcium fluxes). PMID- 2573019 TI - Different modulation of the binding to two phencyclidine (PCP) receptor subtypes: effects of N-methyl-D-aspartate agonists and antagonists. AB - Neurochemical studies have indicated that the dissociative anesthetics, phencyclidine (PCP) and ketamine, act as non-competitive antagonists at the excitatory amino acid, N-methyl-D-aspartate (NMDA), receptor-gated ion channel. Since the binding of PCP and related psychotomimetics, i.e. (+)-N allylnormetazocine [+)-SKF 10047), in mammalian brain is associated with multiple receptor subtypes, their modulation by NMDA agonists and antagonists was investigated. Binding of the potent PCP analog, [3H]PCP-3-OH to the high-affinity sigma/PCP (sigma p) site and (+)-[3H]SKF 10047 to the sigma/haloperidol sensitive (sigma h) site in rat brain membranes was not affected by L-glutamate and NMDA, nor by the competitive NMDA antagonists D-2-amino-5-phosphovaleric acid (AP-5), D 2-amino-7-phosphoheptanoic acid (AP-7). However, binding of [3H]PCP-3-OH to the low-affinity PCP-selective site was enhanced by 4- to 5-fold in the presence of glutamate or NMDA and reduced in a competitive manner by AP-5. The noncompetitive NMDA antagonist, MK-801, was however a potent inhibitor of the binding to both sigma p and PCP sites labeled with [3H]PCP-3-OH. The present results indicate that the high (sigma p) and low-affinity (PCP) sites, that are distinct from the sigma h site, are affected differently by NMDA agonists and antagonists, and thus may represent different receptor domains. PMID- 2573020 TI - Coexistence of varying combinations of neuropeptides with 5-hydroxytryptamine in neurons of the raphe pallidus et obscurus projecting to the spinal cord. AB - The coexistence of varying combinations of substance P (SP), somatostatin (SOM), thyrotropin-releasing hormone (TRH) and met-enkephalin-Arg-Gly-Leu (ENK) with 5 hydroxytryptamine (5-HT) as semiquantitatively revealed by immunocytochemistry in neuronal perikarya of the raphe pallidus et obscurus in the guinea-pig was analyzed. SOM coexisted most frequently with 5-HT, followed by SP, ENK and TRH. Many 5-HT neurons were immunoreactive to 2 or more peptides such as SP/SOM, SOM/ENK, SP/ENK, SOM/TRH, SP/TRH or SOM/SP/ENK. Most of these neurons were shown to project to the spinal cord by retrograde HRP labeling combined with immunocytochemistry. After hemisection of the cervical spinal cord at the C5 level, ENK and 5-HT immunoreactive nerve terminals in the ipsilateral intermediolateral nucleus of the thoracic spinal cord were decreased in number. The results indicate that neurons in the raphe pallidus et obscurus projecting to the spinal cord can be classified into subpopulations according to which peptides coexist with 5-HT, and may have different functions. PMID- 2573021 TI - Dopamine, neuroleptics and reinforced behavior. AB - Animals working for any one of a variety of positive reinforcers (food, water, brain-stimulation...) produce dramatic decrements in operant performance when challenged with dopamine antagonist neuroleptic drugs. This well-established fact has generated considerable research aimed at identifying the precise nature of the drug-induced behavioral impairment. The two most oft-cited hypotheses suggest that much of the reduction in operant responding can be accounted for by either an "anhedonic" or "motoric" consequence of dopamine antagonism. Several novel behavioral paradigms are described which were devised to more clearly elucidate both the motor and reward impairing qualities of neuroleptic agents. Motor performance was assessed in a food-reinforced task in which a computer-operated force-transducer was used to obtain detailed analyses of the biophysical properties of operant responding. These studies suggested that neuroleptics impair the temporal nature (i.e., "timing") of normal operant behavior and not the physical capacity of the animals (i.e., the ability to emit "force"). This identification of a robust "slowing" effect of neuroleptic challenge, suggested that the investigation of putative "reward" impairments should best be conducted with behavioral paradigms in which the test data are collected from undrugged animals. Three such paradigms are described: a partial reinforcement extinction test, an incentive motivation test, and a conditioned place preference test. To date, our results suggest that when motor confounds are avoided, dopamine antagonist drugs can still produce patterns of operant behavior that very closely resemble those observed with actual reductions in reward magnitude. Such data provide support for the contention that central dopaminergic substrates play a role in the neurobiology of positive reinforcement. PMID- 2573022 TI - Receptor subtype-specific dopaminergic agents and conditioned behavior. AB - Dopaminergic neurotransmission has been implicated in reward-related learning. With the advent of pharmacological agents that are relatively specific for D1 and D2 dopamine receptors, it has become possible to assess the role of these receptor subtypes in this form of learning. Antagonist studies have shown that either D1 or D2 receptor blockers produced extinction-like effects on operant responding for food, water or brain stimulation reward and in drug self administration paradigms. They also blocked place preference learning based on amphetamine. Agonist studies showed that D2, but not D1 agonists were self administered, produced place preferences and enhanced responding for conditioned reward. It may be that the dopaminergic signal at the D1 receptor is important for the establishment and maintenance of reward-related learning. From this point of view the effects of D1 antagonists can be understood. D2 antagonists may produce extinction-like effects because they lead to increased dopamine release and, therefore, indirectly mask the dopamine signal at the D1 receptor. D1 agonists may fail to produce reward effects because they, unlike D2 agonists, directly mask the dopaminergic signal at the D1 receptor. PMID- 2573023 TI - Opiate reward: sites and substrates. AB - Opiates appear to have rewarding actions at more than one locus in the brain. Studies of the effects of dopaminergic lesions and dopamine receptor blockade indicate that intravenous heroin self-administration depends importantly on a dopaminergic substrate. Mapping of effective injection sites for morphine conditioned place preference establishes one site of rewarding action near the dopamine cell bodies of the ventral tegmental area (VTA). Studies of the complex interactions of opiates, neuroleptics, and brain stimulation reward confirm that reward-related VTA opioid actions are dopamine-dependent. Opioid injections into the nucleus accumbens (NAS) also facilitate brain stimulation reward and serve as rewards in their own right, though these actions have not yet been localized by identification of negative sites in surrounding regions. The relation of this putative reward site to the dopamine system is not yet clear. Suggestions that the lateral hypothalamus or periaqueductal gray contain opioid reward sites remain to be confirmed. While opioid injections into these sites can be rewarding, these rewarding effects have not been localized to these sites, and opiate injections into each of these areas are reported not to facilitate brain stimulation reward. Intravenous heroin self-administration is not disrupted by kainic acid lesions of the bed nucleus of the lateral hypothalamus. Thus only the VTA and the NAS are firmly established as sites of opiate rewarding actions. Recent reports suggest that the kappa-opioid dynorphin may also have central rewarding actions and central and peripheral aversive actions; the CA3 region of the hippocampus is a possible site of the rewarding action. PMID- 2573025 TI - [A conference with the topic AIDS]. PMID- 2573026 TI - Grief counselling from the mortuary. PMID- 2573024 TI - Cellular investigations of behavioral reinforcement. AB - Using the hippocampal-slice preparation, we attempted to demonstrate operant conditioning of pyramidal cell activity using local micropressure applications of transmitters and drugs as reinforcement; the same injections administered independently of bursting provided a control for direct pharmacological stimulation or facilitation of firing. The results suggested that the spontaneous bursting of individual CA1 pyramidal neurons may be reinforced with activity contingent injections of dopamine and cocaine, whereas, CA3-bursting responses may be reinforced with contingently-applied dynorphin A. We sought to confirm these indications of cellular reinforcement at the behavioral level in studies of hippocampal self-administration (despite the fact that the hippocampus has been ignored as a brain site for chemical self-administration experiments). The results suggested that dynorphin A is a powerful reinforcer of hippocampal self administration behavior when injected in the CA3 field; experiments still in progress suggest that dopamine can reinforce self-administration behavior when injected in the CA1 field. Successful prediction of new behavioral data from operant-conditioning data at the cellular level helps to validate the cellular data by providing suggestive evidence of interrelationship between cellular and behavioral operant conditioning processes. PMID- 2573027 TI - [Unusual complication of foot trauma (tumor-like ossification)]. PMID- 2573028 TI - [Bronchial obstruction exacerbated during beta blocker therapy]. AB - The authors analyse data of 40 patients with chronic obstructive pulmonary disease, in whom the bronchial obstruction made such a severe progress during beta-blocker therapy necessary to admit them to the hospital. Most of them were middle-aged patients with chronic bronchitis, only 8 suffered from bronchial asthma. Because of cardial disorders, usually beta-blockers were given for several days (1/3-rd of the patients received Betaloc, a cardioselective drug), besides 2/3-rd of them received bronchodilator therapy, too. In spite of this, all patients needed hospitalize because of dyspnoea, obstruction, and hypoxaemia in 1/3-rd. Although deterioration of their condition has resulted, beta-blocker wasn't discontinued by the medical attendant, notwithstanding chronic obstructive pulmonary disease was known previously in 33 patients. PMID- 2573029 TI - Activation of an EGFR/neu chimeric receptor: early intracellular signals and cell proliferation responses. AB - Two clones of NIH3T3 fibroblasts, NEN37 and NEN7, overexpressing chimeric EGF/neu receptors (3 x 10(5) and 1 x 10(6) receptors/cell, respectively), were treated with EGF in order to identify the array of intracellular signals generated after activation of the neu proto-oncogene product. The results thus obtained were correlated with the effects of EGF on cell growth, investigated by both [3H]thymidine incorporation and long term (5 days) proliferation studies. In addition to the stimulation of the neu tyrosine kinase, previously reported by Lehvaslaiho et al. (EMBO J., 8, 159-166, 1989), EGF (10(-9)-10(-8) M) was found to induce marked increases of both [Ca2+]i and plasma membrane potential (investigated by the fura-2 and bis-oxonol techniques) which, in their initial phase, were only marginally dependent on the presence of Ca2+ in the incubation medium. These responses were inhibited, but only in part (40-50%) by phorbol ester activators of protein kinase C. Moreover, inositolphosphate analysis (by anion exchange chromatography) revealed hydrolysis of membrane polyphosphoinositides. All these effects of EGF were more prompt and much larger in NEN7 than NEN37 cells. The EGF concentration-dependence curves (measured by both [3H]thymidine incorporation and long-term proliferation assay) were quite different in the two cell clones. In the cells expressing the lower number of receptors measurable growth stimulation was observed at 10(-10), and maximal effect at 10(-9) M EGF. In NEN7 cells the curve was much more shallow, with measurable stimulation already at 10(-12) and maximal effect at 10(-8) M EGF. The maximal growth effect was approximately the same for the two cell clones. It is concluded that the intracellular signals identified here may play a limited role in the neu-induced cell proliferation, but are possibly involved in the acquisition of the tumoral phenotype typically expressed by the EGF-treated NEN7 cells. PMID- 2573030 TI - Antinociceptive effects of the novel anxiolytic buspirone in three pain tests in rats. AB - The analgesic efficacy of the novel anxiolytic buspirone in tests of acute thermal, mechanical and formalin-induced chemical-inflammatory pain were examined. Buspirone produced dose- and time-dependent analgesia in all nociceptive tests, with greatest effects against chemical and mechanical pain. Buspirone was less potent in the thermal pain test. Locomotor and overt behavioral activities were unaffected at any dose tested. These data suggest the potential use of buspirone in the management of specific types of pain. PMID- 2573031 TI - Implications of genetic exchange in the study of protozoan infections. AB - Genetic exchange is now known to occur during the life-cycle of many parasitic protozoa, including malaria parasites, coccidia and trypanosomes. The process is studied by making deliberate crosses between cloned organisms differing in clearly defined markers. In malaria parasites, crosses have been made between parasites differing in characters such as isoenzymes, antigens and other proteins, drug sensitivity, and chromosome and other DNA polymorphisms. Crosses are made by transmitting a mixture of gametes of each clone through mosquitoes to allow cross-fertilization to take place, and examining the resulting progeny by cloning for organisms exhibiting non-parental combinations of characters. The inheritance of many characters, such as antigen and protein variants, is in accordance with Mendelian expectations for a haploid organism. Recombination occurs at a higher than expected frequency. Studies on chromosomes have show that crossing-over events commonly occur following meiosis of hybrid zygotes. Repetitive DNA and subtelomeric regions of chromosomes appear to be particularly susceptible to such recombination events. In trypanosomes, crosses between clones of Trypanosoma brucei have shown that hybrids are formed during tsetse fly transmission. The organism appears to be mainly diploid, but some characters including certain chromosomes seem to be inherited in a non-Mendelian manner. PMID- 2573032 TI - Prenatal diagnosis of genetic disorders by DNA analysis. PMID- 2573033 TI - Adherence kinetics of Haemophilus influenzae type b to eucaryotic cells. AB - Haemophilus influenzae type b (Hib) adhere in vitro to buccal epithelial cells (BEC) via pili (fimbriae). In vivo studies have conflicting lines of evidence concerning the role of pili in adherence. This study characterizes the kinetics of Hib binding to BEC and compares pilus versus nonpilus adherence. Adherence was assessed using radioactive, immunofluorescence, and electron microscopic techniques. Paired Hib strains were obtained from three children with Hib meningitis; piliated Hib were isolated from the nasopharynx and nonpiliated isolates from the cerebrospinal fluid. Piliated strains adhered avidly to BEC after a 1-h incubation, whereas nonpiliated strains adhered poorly (p = 0.002). Kinetic analysis revealed increased adherence to BEC by piliated and nonpiliated strains with time, with maximum adherence occurring between 24-36 h. Immunofluorescence and electron microscopic assays visually confirmed the radioactive adherence findings. These results characterize the kinetics of Hib adherence to BEC. BEC incubated with nonpiliated Hib for 30 h were found to be coated with piliated strains, suggesting the induction of pilus production or the selection of a piliated subpopulation. PMID- 2573035 TI - [Beta 2 adrenergic and synthetic atropine drugs as inhalants in asthma in children]. AB - Inhaled medication is a well-known treatment for asthma in childhood, because it is rapidly effective, in low doses, and without resultant side effects. The usual methods of aerosol formation are nebulization, metered-dose inhaler and spacers. We have reviewed the standard devices available and their use with sympathomimetic agents and synthetic atropines. PMID- 2573034 TI - Triacylglycerol and glycogen contents in the human gastric mucosa: effect of histamine H2, muscarinic and gastrin receptors blockade. AB - Triacylglycerol and glycogen contents in the gastric mucosa were studied in 20 patients with duodenal ulcer before and after two-week treatment with cimetidine, pirenzepine or proglumide. Determinations of both metabolic substrates were performed in mucosal slices taken from gastric corpus and antrum during endoscopic examination: the results were expressed per mg of protein. Concentrations of triacylglycerol were significantly elevated after treatment with any of the blocking agents: in the gastric corpus by 88% after cimetidine application, 69% after pirenzepine and 102% after proglumide; in the antrum by 76, 59 and 132%, respectively. No significant changes in the mucosal contents of glycogen were observed. It is concluded that an increase in the mucosal concentration of triacylglycerol can be connected with an inhibition of the gastric acid secretion following treatment with receptor-blocking drugs. PMID- 2573036 TI - [Undescended testis--a review]. AB - According to registrations in several countries far too many retractile testes are falsely included in patient materials of truly undescended testes. The problem of overtreatment is so great that even with a low frequency of complications to the treatment, the netto effect on the population's fertility may become negative. The most important task for the clinician is to discern truly undescended from retractile testes. The decision has to be based on thorough clinical examination, but ultrasonography may provide objective evidence. The article presents a state-of-the-art review on experimental and clinical aspects of cryptorchidism. PMID- 2573037 TI - Detection of minority point mutations by modified PCR technique: a new approach for a sensitive diagnosis of tumor-progression markers. AB - The detection of point mutations correlated with diseases, in enzymatically amplified DNA sequences (Polymerase Chain Reaction), is currently performed by digestion of PCR products when an existing restriction site disappears at least in one allele of the amplified mutated sequence or by allele specific radiolabeled probes in all other cases. These methods are the most sensitive but they cannot detect a mutation if it is present in less than 5% of the studied cells. We describe here a method based on the introduction of an artificial restriction site, using a modified primer during the PCR, which creates a RFLP indicative of the studied mutation. This RFLP is detected by a radiolabeled oligonucleotide probe which is not related to the mutation. Our approach multiplies the sensitivity by a factor of 1000 and it is practical for use in screening purposes and the detection, after treatment, of the residual disease in human malignancies. Using this method we detected 20% more mutations at codon 12 in the Ki ras oncogene in DNA from colorectal cancers that were undetectable with all the previous methods. PMID- 2573038 TI - Direct haplotyping of chromosomal segments from multiple heterozygotes via allele specific PCR amplification. PMID- 2573039 TI - The anonymous probe pR1-4 which identifies the locus D13S59 detects a BanII RFLP. PMID- 2573040 TI - The anonymous probe pF5A identifying the locus D13S61 detects RFLPs with XmnI and BanII. PMID- 2573041 TI - The single copy probe pG24E2.4 [D13S21] reveals a Bsp1286 RFLP at 13q14.1-q14.2. PMID- 2573042 TI - The anonymous probe pG50 identifying the locus D13S24 detects a two allele RFLP with SspI. PMID- 2573043 TI - The alpha chain of human propionyl CoA carboxylase (PCCA) mapped to chromosome 13) detects an RFLP with XmnI. PMID- 2573044 TI - Isolation and mapping of a polymorphic DNA sequence (CJ52.161) on chromosome 16 [D16S150]. PMID- 2573046 TI - [Giant cell arteritis]. AB - Giant cell arteritis are characterized by histologic disruption features of the lamina elastica of large arteries with mononuclear cells and giant cells infiltration. Systemic giant cell arteritis, temporal arteritis and Takayasu's arteritis are included in this group. Takayasu's arteritis predominantly affects the teenage girl and involves the aorta and its mayor branches. Early manifestations are systemic; later appears signs and symptoms of occlusion. PMID- 2573045 TI - Localization of monoamine oxidase A and B genes on the mouse X chromosome. PMID- 2573047 TI - [Polyarteritis]. AB - Polyarteritis is an uncommon childhood disease. The different forms of polyarteritis are described. Polyarteritis nodosa is a rare multisystemic disease characterized by high fever, calf pain and subcutaneous nodules. Infantile polyarteritis nodosa shows a peculiar predilection for the coronary arteries and has the worst prognosis. Polyarteritis cutaneous is the localized and mild form of polyarteritis. Hypersensitivity angiitis is described as a leukocytoclastic vasculitis identified by palpable purpura. PMID- 2573048 TI - A retrospective analysis of fenoldopam renal excretion in 65 subjects: evidence for possible intrarenal formation of fenoldopam from its metabolites. AB - Clinical studies have suggested that the dopamine DA1 agonist, fenoldopam, may exhibit nonlinear renal excretion in humans. A retrospective population pharmacokinetic analysis of the renal excretion of fenoldopam and one of its major metabolites, fenoldopam-8-sulfate, was conducted in 65 healthy volunteers to examine this phenomenon. Fenoldopam-8-sulfate exhibited a mean (+/- SE) renal plasma clearance of 129 +/- 4 ml/min, which was independent of its AUC. In contrast, fenoldopam renal plasma clearance ranged from 2220 to 150 ml/min and decreased nonlinearily with increasing fenoldopam AUC. Fenoldopam renal clearance was characterized as a function of fenoldopam AUC using a nonlinear saturation model. The analysis predicted an initial maximal renal clearance of 2852 ml/min, which decreased to 78 ml/min at maximal inhibition. The fenoldopam AUC required to half-saturate fenoldopam renal clearance was 5.2 ng x hr/ml. The elevated clearance values for fenoldopam, beyond normal physiologic limits for renal blood flow in man, suggest that intrarenal formation of fenoldopam from one or more of its circulating metabolites may be contributing to the observed nonlinear decreases in fenoldopam renal excretion. Preliminary data from our laboratory suggest that in vivo desulfation of fenoldopam-8-sulfate to fenoldopam does occur in the dog. PMID- 2573049 TI - A comparison of the cholinergic activity of selected H2-antagonists and sulfoxide metabolites. AB - Famotidine and selected H2-antagonists were evaluated with respect to toxicity and selected pharmacological activities. When administered intraperitoneally to mice at a dose equivalent to 10 times their respective H2-antagonist ED50 values, no deaths were observed. Similarly, no alteration in brain ACh concentrations or overt pharmacological effects were noted. However, at 400 mg/kg, ranitidine produced 89% lethality, followed by cimetidine (11%) and famotidine. Only cimetidine and famotidine at this dose significantly elevated brain acetylcholine levels. These results do not correlate with the in vitro data, where ORF-17578 and ranitidine were the most potent entities with respect to acetylcholinesterase inhibition (approximately 1-2 X 10(-6) M), followed by nizatidine greater than cimetidine greater than famotidine. The sulfoxide metabolites of ranitidine and cimetidine were approximately one-tenth as potent as their parent compounds with respect to inhibition of acetylcholinesterase. Direct muscarinic stimulation or potentiation of acetylcholine-induced contraction in ileal tissue was not observed for any of the H2-antagonists. PMID- 2573051 TI - Inhibitory effects of sulfasalazine and related compounds on superoxide production by human polymorphonuclear leukocytes. AB - The inhibitory effects of sulfasalazine, some sulfasalazine-related compounds and indomethacin on superoxide production by human polymorphonuclear (PMN) leukocytes were studied. The inhibition of the chemotactic peptide (FMLP)-induced superoxide production, which is membrane receptor-mediated, was strongly dependent on the concentration both of the secretory stimulus and of the test compounds, indicating an interaction between the receptor and the test compound. Furthermore, a positive correlation was found between the lipophilicity of the compound and the degree of inhibition. However, when the receptor was by-passed by direct activation of the receptor-linked G protein by the use of fluoride ions as secretory stimuli, the test compounds still inhibited superoxide production. On the other hand, superoxide production by cells stimulated with phorbol ester was not inhibited by the test compounds. Furthermore, the production of phosphatidic acid was decreased in the presence of sulfasalazine, indicating impaired phosphoinositide metabolism. The inhibition of this metabolism was not due to increased intracellular concentrations of cyclic AMP, although sulfasalazine did inhibit cyclic nucleotide phosphodiesterase. We conclude that sulfasalazine attenuates superoxide production by PMN leukocytes at a post receptor site of action at a step before the activation of protein kinase C, possibly by interfering with the phosphoinositide metabolism but independent of cyclic AMP. PMID- 2573050 TI - Preferential inhibition of mouse hepatic coumarin 7-hydroxylase by inhibitors of steroid metabolizing monooxygenases. AB - Etomidate, metomidate and metyrapone, all potent inhibitors of steroid metabolizing monooxygenases, inhibit preferentially coumarin 7-hydroxylase (COH) amongst several liver microsomal monooxygenase activities from control and pyrazole-treated D2 mice in vitro. SKF-525A, an inhibitor of phenobarbital inducible monooxygenase activities has a much weaker effect on COH than the other three drugs, even though COH is a phenobarbital-inducible enzyme. Treatment of mice with eto- and metomidate decreases the microsomal COH also in vivo while the other activities remained unchanged (with the exception of 7-ethoxycoumarin O deethylase (ECDE) in case of metomidate). Despite of the decrease in COH no parallel decrease in the amount of microsomal P450Coh (P450 isoenzyme highly active in the 7-hydroxylation of coumarin) could be found in dot immuno-binding analysis. These data suggest that among several liver microsomal P450 isoenzymes, metyrapone, eto- and metomidate interact preferentially with the P450Coh and that eto- and metomidate may alter selectively the catalytic properties of P450Coh leading to decreased enzyme activity. Two different Ks-values could be found for all three drug in their binding to microsomal cytochrome(s) P450. Based on substrate binding spectra, potassium ferricyanide treatment does not dissociate the complex between reduced P450 and metomidate and does it only partly for etomidate. Furthermore potassium ferricyanide treatment of microsomes does not increase COH after in vivo treatment of mice with eto- and metomidate. These data further suggest that the complex between P450Coh and eto- and metomidate may be particularly strong and independent from the redox state of the haem iron. PMID- 2573052 TI - Adaptive changes in alpha-2 adrenoceptor mediated responses: analgesia, hypothermia and hypoactivity. AB - The acute effects of the alpha-2 adrenoceptor agonists, clonidine and guanfacine, upon antinociception, hypothermia and motor activity were compared under conditions of receptor antagonism, denervation, and chronic administration of a tricyclic antidepressant compound. The analgesic actions of clonidine and guanfacine were antagonised by idazoxan, an alpha-2 receptor antagonist, but potentiated by pretreatment with the noradrenaline neurotoxin DSP4, and attenuated by chronic treatment with desipramine (DMI). Clonidine- and guanfacine induced hypothermia was antagonised by idazoxan, potentiated by prior treatment with DSP4 and attenuated by chronic administration with DMI. Both clonidine and guanfacine produced decreases in motor activity that were attenuated by idazoxan but unaffected by prior DSP-4 treatment. Chronic DMI administration also attenuated clonidine-induced hypoactivity but potentiated guanfacine-induced hypoactivity. These diverse results describe both similar and differential adaptive mechanisms modulating the functional effect of alpha-2 receptor systems in the central nervous system. PMID- 2573053 TI - [Suppressor genes in the malignant transformation and genic deletions in human cancers]. AB - Experimental studies of cell malignant transformation have shown that such transformation requires the alteration of genes that can control the expression of the transformed phenotype. In human oncology, the role of genes that suppress malignant transformation is suggested by the fact that specific genic deletions have been detected by polymorphism-detecting probes in many malignant tumours. The only oncosuppressor gene that is characterized at the moment is the inactivated RB gene in retinoblastoma. Molecular biology studies have shown that in retinoblastoma and other malignant tumours this gene is altered in its structure or expression. In addition to retinoblastoma, nephroblastoma in children and colonic carcinoma constitute models for the study of oncosuppressor genes in human oncology. PMID- 2573054 TI - [Neuropathies in vasculitis]. PMID- 2573056 TI - Interaction of glucagon-like peptide-1(7-36)amide and somatostatin-14 in RINm5F cells and in the perfused rat pancreas. AB - Glucagon-like peptide-1(7-36)amide [GLP-1(7-36)amide], a new important incretin candidate, binds to specific high-affinity receptors on rat insulinoma-derived beta-cells (RINm5F). In the present study, the effect of somatostatin-14 on the GLP-1(7-36)amide-induced insulin release and cAMP generation in this cell line was investigated. Somatostatin did not decrease basal insulin release of RINm5F cells. The GLP-1(7-36)amide-induced insulin release was decreased concentration dependently by somatostatin. Somatostatin, 1 microM reduced the maximally GLP-1(7 36)amide-stimulated (0.1 microM) insulin release to basal insulin levels. The GLP 1(7-36)amide-induced cAMP production was significantly decreased by somatostatin in a concentration-dependent manner. The GLP-1(7-36)amide concentration causing half-maximal cAMP production was 2.98 +/- 1.56 nM. Somatostatin left the EC50 unaltered but decreased the maximal GLP-1(7-36)amide effect for 32% in the presence of 1 nM somatostatin and for 50% at 1 microM. In additional experiments, the interaction of both hormones was evaluated in the perfused pancreas as a nontumor model. Somatostatin (1 nM, 1 microM) inhibited the glucose-induced (6.7 mM) and GLP-1(7-36)amide-potentiated (0.05, 0.5, and 5 nM) insulin release dose dependently. The biphasic pattern of insulin release remained preserved. The GLP 1(7-36)amide-induced insulin release is potently inhibited by somatostatin-14. This effect was demonstrated in different model systems for beta-cell function studies. The present data allow the conclusion that the somatostatin action upon GLP-1(7-36)amide effects is at least partly related to regulation of intracellular cyclic nucleotides. PMID- 2573055 TI - Membrane receptors for peptides in experimental and human pancreatic cancers. AB - Membrane receptors for [D-Trp6]-luteinizing hormone-releasing hormone [( D-Trp6] LH-RH), somatostatin (SS-14), and epidermal growth factor (EGF) were investigated in experimental N-nitrosobis-(2-oxopropyl)-amine (BOP)-induced pancreatic cancers of hamsters and in specimens of normal human pancreas and human pancreatic cancer obtained from autopsies. Membrane receptors for [D-Trp6]-LH-RH were absent in the pancreas of normal hamsters, but appeared after the carcinoma was induced with BOP. Binding capacity of SS-14 receptors was lower in membranes of BOP-induced pancreatic cancers than in the normal pancreas. In the BOP-induced pancreatic cancers, the receptors were also characterized following in vivo treatment of hamsters with microcapsules of the agonist [D-Trp6]-LH-RH, somatostatin analog RC 160, and the combination of both peptides, which resulted in significant tumor inhibition. Therapy with [D-Trp6]-LH-RH and RC-160, alone or in combination, decreased the binding capacity of receptors for [D-Trp6]-LH-RH, but increased Bmax for SS-14. There were no significant changes in characteristics of the EGF receptor following these therapies. Membranes from human pancreatic cancers showed binding sites for [D-Trp6]-LH-RH, but no binding was detected in normal human pancreas. The presence of receptors for LH-RH in pancreatic tumors of hamster and humans raises the intriguing possibility that LH-RH could be involved in complex interactions that contribute to the appearance of pancreatic cancer. The binding capacity of receptors for SS-14 in human pancreatic cancer membranes was lower, while Bmax for EGF was higher, as compared to normal pancreas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573057 TI - Somatostatin infused during acute pancreatitis retains its biological activity. AB - Somatostatin (SST) is used in the treatment of acute pancreatitis (AP) to inhibit pancreatic exocrine secretion, which represents one of the goals of medical treatment in this disease. Its therapeutic efficacy, however, is poor. One hypothesis, which has not yet been investigated, is that i.v. SST might be broken down by blood proteolytic enzymes. In order to evaluate the structural integrity and biological activity of infused SST, somatostatin-like immunoreactivity (SLI) and levels of pancreatic enzymes were monitored in the blood stream during the infusion of SST-14 (3,5 micrograms/kg/h for 48 h) in eight patients with severe acute pancreatitis. SLI was measured by both radioimmunoassay (RIA) and high pressure liquid chromatography (HPLC). The results indicate that SLI levels increase promptly after the beginning of infusion, with a slower increase between 6 and 36 h, and a rapid increase again at 48 h. HPLC analysis shows a single peak of SLI with the same retention time as standard SST-14. Total amylase, lipase, and trypsinogen significantly decreased compared with pretreatment values (48, 63.1, and 77.4%, respectively) after 24 h of SST infusion, while a decrease in elastase 1 (62.6%) was observed later at 48 h. These results indicate that in severe AP, somatostatin recovered in plasma retains its biological activity: it inhibits pancreatic circulating enzymes, an action not influenced by breakdown of the peptide, as demonstrated by HPLC of the SLI measured in plasma. PMID- 2573058 TI - Drosophila NK-homeobox genes. AB - Four Drosophila melanogaster homeobox genes were found by screening a genomic DNA library with oligodeoxynucleotides that correspond to a conserved amino acid sequence that is part of the putative of homeobox proteins that recognizes nucleotide sequences in DNA. The amino acid sequences of NK-2, NK-3, and NK-4 homeoboxes are more closely related to one another (59-66% homology) than they are to other Drosophila homeoboxes (28-54% homology), whereas the homeobox of NK 1 is most closely related, in order of decreasing homology, to muscle segment homeobox, zerknullt-1, NK-3, and distal-less homeoboxes. Three of the genes, NK 1, NK-3, and NK-4, comprise a cluster of homeobox genes located in the 93E1-5 region of the right arm of the third chromosome, whereas the fourth homeobox gene, NK-2, is located in the 1C1-5 region of the X chromosome. PMID- 2573059 TI - Individuals with IgA deficiency and common variable immunodeficiency share polymorphisms of major histocompatibility complex class III genes. AB - IgA deficiency and common variable immunodeficiency are heritable disorders that can occur within the same family. Both immunodeficiencies are characterized by arrests in B-cell differentiation that vary in the extent of the immunoglobulin isotypes involved. A high frequency of major histocompatibility complex supratypes associated with a null allele of the gene encoding the C4A isotype of complement component C4 has been observed in IgA-deficient individuals. In search of a genetic linkage between the two immunodeficiencies, we examined the major histocompatibility complex (MHC) class III genes encoding complement components C2, C4A, and C4B and steroid 21-hydroxylase in addition to the HLA serotypes in individuals with either common variable immunodeficiency or IgA deficiency. Twelve of 19 patients with common variable immunodeficiency (63%, P less than 0.001) and 9 of 16 patients with IgA deficiency (56%, P less than 0.01) had rare C2 alleles and/or C4A and 21-hydroxylase A deletions, whereas these gene features were seen in only 5 of 34 healthy individuals (15%) in the control group. Nine of 11 patients with C4A deletion had an HLA haplotype consistent with the MHC supratype HLA-A1, Cw7, B8, C4AQ0, C4B1, BfS, DR3 previously found to be associated with IgA deficiency. The data support the hypothesis that common variable immunodeficiency and IgA deficiency are related disorders, susceptibility to which is determined by a gene(s) within or near the MHC class III gene region on chromosome 6. PMID- 2573061 TI - Convergence and divergence of neurotransmitter action in human cerebral cortex. AB - The postsynaptic actions of acetylcholine, adenosine, gamma-aminobutyric acid, histamine, norepinephrine, and serotonin were analyzed in human cortical pyramidal cells maintained in vitro. The actions of these six putative neurotransmitters converged onto three distinct potassium currents. Application of acetylcholine, histamine, norepinephrine, or serotonin all increased spiking by reducing spike-frequency adaptation, in part by reducing the current that underlies the slow after hyperpolarization. In addition, application of muscarinic receptor agonists to all neurons or of serotonin to middle-layer cells substantially reduced or blocked the M-current (a K+ current that is voltage and time dependent). Inhibition of neuronal firing was elicited by adenosine, baclofen (a gamma-aminobutyric acid type B receptor agonist), or serotonin and appeared to be due to an increase in the same potassium current by all three agents. These data reveal that individual neuronal currents in the human cerebral cortex are under the control of several putative neurotransmitters and that each neurotransmitter may exhibit more than one postsynaptic action. The specific anatomical connections of these various neurotransmitter systems, as well as their heterogeneous distribution of postsynaptic receptors and responses, allows each to make a specific contribution to the modulation of cortical activity. PMID- 2573060 TI - Dopamine- and cAMP-regulated phosphoprotein (DARPP-32) and dopamine DA1 agonist sensitive Na+,K+-ATPase in renal tubule cells. AB - The cellular localization of DARPP-32, a dopamine- and cAMP-regulated phosphoprotein of Mr 32,000 that appears to mediate certain actions of dopamine in the mammalian brain by acting as an inhibitor of protein phosphatase 1, was studied in the kidney of several species. DARPP-32 mRNA and DARPP-32-like immunoreactivity were found in the cytoplasm of cells in the thick ascending limb of the loop of Henle. The specific dopamine DA1 agonist SKF 82526 caused a dose dependent inhibition of Na+,K+-ATPase activity, which could be blocked by SCH 23390, a specific DA1 antagonist, and by PKI-(5-24) amide, a specific inhibitor of cAMP-dependent protein kinase. The results indicate that DA1 dopamine receptors and DARPP-32, an intracellular third messenger for dopamine, are part of the signal-transduction process for dopamine acting on renal tubule cells. PMID- 2573062 TI - Crystal structure of the Glu-239----Gln mutant of aspartate carbamoyltransferase at 3.1-A resolution: an intermediate quaternary structure. AB - The structure of the unligated Glu 239----Gln mutant of Escherichia coli aspartate carbamoyltransferase (EC 2.1.3.2) has been determined to 3.1-A resolution and refined to a crystallographic residual of 0.22 in the space group P321. The unit-cell dimensions of the unligated enzyme are a = 122.3 A, c = 147.1 A. The c axis cell length is intermediate between the c axis lengths of the T (tense)(c = 142.2 A) and R (relaxed) (c = 156.2 A) state structures. Furthermore, the quaternary structure of the mutant enzyme is intermediate between the quaternary structures of the T form and the R form. The differences between the quaternary structures of the Glu-239----Gln and T-form enzymes can be described as follows: the separation between the catalytic trimers increases by approximately 1.5 A along the threefold axis, and they each rotate in opposite directions approximately 0.5 degree around the threefold axis, whereas the regulatory dimers rotate approximately 2 degrees around the twofold axes. PMID- 2573063 TI - Glutamic acid-113 serves as the retinylidene Schiff base counterion in bovine rhodopsin. AB - The characteristic wavelength at which a visual pigment absorbs light is regulated by interactions between protein (opsin) and retinylidene Schiff base chromophore. By using site-directed mutagenesis, charged amino acids in bovine rhodopsin transmembrane helix C were systematically replaced. Substitution of glutamic acid-134 or arginine-135 did not affect spectral properties. However, substitution of glutamic acid-122 by glutamine or by aspartic acid formed pigments that were blue-shifted in light absorption (lambda max = 480 nm and 475 nm, respectively). While the substitution of glutamic acid-113 by aspartic acid gave a slightly red-shifted pigment (lambda max = 505 nm), replacement by glutamine formed a pigment that was strikingly blue-shifted in light absorption (lambda max = 380 nm). The 380-nm species existed in a pH-dependent equilibrium with a 490-nm species such that at acidic pH all of the pigment was converted to lambda max = 490 nm. We conclude that glutamic acid-113 serves as the retinylidene Schiff base counterion in rhodopsin. We believe that this opsin chromophore interaction is an example of a general mechanism of color regulation in the visual pigments. PMID- 2573065 TI - Isolation and characterization of a 60-residue intestinal peptide structurally related to the pancreatic secretory type of trypsin inhibitor: influence on insulin secretion. AB - We have isolated from pig intestine a 60-residue polypeptide initially identified by its inhibition of glucose-induced insulin secretion from perfused pancreas. The amino acid sequence of this porcine polypeptide was determined and found to be markedly similar to that of the pancreatic secretory trypsin inhibitor (41% residue identities). Furthermore, the disulfide arrangements of these two proteins appear identical, suggesting related overall conformations. However, the polypeptide, now named PEC-60 (peptide with N-terminal glutamic acid, C-terminal cysteine, and a total of 60 residues), was found not to inhibit trypsin. The amino acid sequence is also similar to that of a peptide recently isolated from rat bile/pancreatic juice which stimulates the release of cholecystokinin. The biological role of PEC-60 is not known, but the effect on insulin secretion and the homologies observed suggest important biological activities and interesting structural relationships. PMID- 2573066 TI - Retrovirus-mediated gene transfer to purified hemopoietic stem cells with long term lympho-myelopoietic repopulating ability. AB - Despite recent advances in marrow stem cell purification, controversy about the nature and heterogeneity of cells with the potential for long-term repopulation of lymphoid and myeloid tissues remains. Essential to the resolution of these questions is the use of strategies to track the progeny produced in vivo from individual hemopoietic stem cells in purified populations. We have used a procedure for obtaining highly enriched populations of stem cells with competitive repopulating ability from male mice (pretreated with 5-fluorouracil), and in this paper we present the results of studies in which small numbers (150 2000) of these cells were exposed to supernatant containing a helper-free recombinant retrovirus carrying the neomycin-resistance gene and then were transplanted together with 2 x 10(5) "compromised" female marrow cells into irradiated female recipients. Male cells--i.e., progeny of purified stem cells- were found in one or more of the tissues examined (peripheral blood, marrow, spleen, and thymus) in 28 of 28 mice evaluated at various times between 35 and 196 days after transplantation. In 20 of these mice (71%), the neomycin resistance gene was also detected, although not always at a level that correlated with the proportion of male cells. Analysis of spleen colonies (day 12) generated in secondary recipients confirmed that viral integration was confined to male repopulating cells. In three mice direct evidence of a common clone in both lymphoid and myeloid tissues was also obtained. These results show the feasibility of retrovirus-mediated gene transfer to highly purified populations of lympho-myelopoietic stem cells with long-term (6 months) repopulating potential by using a supernatant infection protocol. This approach should facilitate further analysis of hemopoietic stem cell control in vivo and find future applications in the evolving use of bone marrow transplantation for hemopoietic rescue and gene therapy. PMID- 2573064 TI - Lineage-restricted expression of homeobox-containing genes in human hematopoietic cell lines. AB - We investigated the role of homeobox-containing genes in human hematopoiesis because homeobox genes (i) control cell fate in the Drosophila embryo, (ii) are expressed in specific patterns in human embryos, and (iii) appear to function as transcription factors that control cell phenotype in other mammalian organs. Using four homeobox probes from the HOX2 locus and a previously undescribed homeobox cDNA (PL1), we screened mRNAs from 18 human leukemic cell lines representing erythroid, myeloid, and T- and B-cell lineages. Complex patterns of lineage-restricted expression are observed: some are restricted to a single lineage, while others are expressed in multiple lineages. No single homeobox gene is expressed in all types of hematopoietic cells, but each cell type exhibits homeobox gene expression. HOX2.2 and -2.3 homeobox-containing cDNAs were cloned from an erythroleukemia cell (HEL) cDNA library, while the homeobox cDNA PL1 was isolated from a monocytic cell (U-937) library. Differentiation of HEL and K-562 cells with various inducers results in modulation of specific homeobox transcripts. In addition, HOX2.2 is expressed in normal bone marrow cells. We have demonstrated (i) lineage-restricted expression of five homeobox genes in erythroid and monocytic cell lines; (ii) expression of additional homeobox genes in other cell lineages (HL-60 and lymphoid cells); (iii) expression of one homeobox gene in normal marrow cells; and (iv) modulation of expression during differentiation. These data suggest that these genes play a role in human hematopoietic development and lineage commitment. PMID- 2573067 TI - Loss of heterozygosity suggests tumor suppressor gene responsible for primary hepatocellular carcinoma. AB - Primary hepatocellular carcinoma (PHC), epidemiologically associated with chronic hepatitis B virus (HBV) infection, has historically been felt to be caused by the activation or introduction of an oncogene. However, transforming sequences from human PHC have not been reproducibly isolated. In this paper, evidence is presented that suggests PHC may result instead from the loss of an anti-oncogene. Seven of 12 human primary liver tumors tested against a panel of restriction fragment length polymorphisms (RFLPs) demonstrated loss of constitutional heterozygosity for markers on chromosome 4. Tumor and nontumor liver tissue were typed for 11 chromosome 4 RFLPs. In addition, at least one RFLP on nine other chromosomes (1, 2, 6, 7, 9, 11, 13, 14, and 17) was tested for allelic loss. Seven of nine tumors constitutionally heterozygous for chromosome 4q markers showed allele loss in tumor tissue. Six of the seven samples were jointly informative for both 4p and 4q markers. Five of the six demonstrated loss for only 4q RFLPs. In one individual, in which two samples were taken from distant locations within the same tumor, both samples showed loss of the same alleles. Among the other chromosomes informative for allele loss, one tumor showed changes on 13q. No other changes were observed in RFLPs located on the eight other chromosomes tested. These results indicate that an anti-oncogene may be located on 4q and suggest a mechanism for PHC and other cancers seroepidemiologically related to virus infection. Liver cancer caused by chronic HBV infection or other environmental agents may be linked through genetic events responsible for the loss of a tumor suppressor locus (anti-oncogene) located on chromosome 4. PMID- 2573068 TI - Long-term expression of human adenosine deaminase in mice transplanted with retrovirus-infected hematopoietic stem cells. AB - Long-term stable expression of foreign genetic sequences transferred into hematopoietic stem cells by using retroviral vectors constitutes a relevant model for somatic gene therapy. Such stability of expression may depend on vector design, including the presence or absence of specific sequences within the vector, in combination with the nature and efficiency of infection of the hematopoietic target cells. We have previously reported successful transfer of human DNA encoding adenosine deaminase (ADA) into CFU-S (colony-forming unit spleen) stem cells using simplified recombinant retroviral vectors. Human ADA was expressed in CFU-S-derived spleen colonies at levels near to endogenous enzyme. However, because of the lack of an efficient dominant selectable marker and low recombinant viral titers, stability of long-term expression of human ADA was not examined. We report here the development of an efficient method of infection of hematopoietic stem cells (HSC) without reliance on in vitro selection. Peripheral blood samples of 100% of mice transplanted with HSC infected by this protocol exhibit expression of human ADA 30 days after transplantation. Some mice (6 of 13) continue to express human ADA in all lineages after complete hematopoietic reconstitution (4 months). The use of recombinant retroviral vectors that efficiently transfer human ADA cDNA into HSC leading to stable expression of functional ADA in reconstituted mice, provides an experimental framework for future development of approaches to somatic gene therapy. PMID- 2573069 TI - Production of human glucocerebrosidase in mice after retroviral gene transfer into multipotential hematopoietic progenitor cells. AB - The human glucocerebrosidase (GC) gene has been transferred efficiently into spleen colony-forming unit (CFU-S) multipotential hematopoietic progenitor cells, and production of human GC RNA and protein has been achieved in transduced CFU-S colonies. High-titer retroviral vectors containing the human GC cDNA were constructed. Mouse bone marrow cells were stimulated with hematopoietic growth factors, infected by coculture with producer cells, and injected into lethally irradiated animals. Four vectors were compared with respect to gene-transfer efficiency into CFU-S progenitors. One vector (G vector) required high concentrations of interleukins 3 and 6 during stimulation and coculture for efficient transduction of CFU-S progenitors. The remaining three vectors (NTG, GTN, and GI vectors) transduced these progenitors at infection frequencies approaching 100% using low concentrations of hematopoietic growth factors to stimulate cell division prior to and during the infection. Vectors using the viral long terminal repeat enhancer/promoter to drive the human GC cDNA produced high levels of human GC RNA in the progeny of CFU-S progenitors after gene transfer. When an internal herpes simplex thymidine kinase promoter assisted by a mutant polyoma enhancer was used to drive the human GC cDNA (NTG vector), little or no human GC RNA was detected in transduced CFU-S colonies. All three vectors producing human GC RNA in CFU-S colonies can generate human GC as detected by immunochemical analysis of CFU-S colonies. NTG vector-infected bone marrow cells were transplanted into W/Wv recipients to generate long-term reconstituted mice. The capacity of the viral long terminal repeat and the internal thymidine kinase promoter to direct synthesis of RNA in transduced bone marrow and spleen cells 5 months after bone marrow transplantation reflected the performance of these promoters in NTG-transduced CFU-S colonies. PMID- 2573070 TI - Germ-line transmission of a planned alteration made in a hypoxanthine phosphoribosyltransferase gene by homologous recombination in embryonic stem cells. AB - Embryonic stem cells (derived from 129/Ola mice) containing a mutant hypoxanthine phosphoribosyltransferase gene that had been corrected in vitro in a planned manner by homologous recombination were injected into blastocysts obtained from C57BL/6J mice. The injected blastocysts were introduced into pseudopregnant female mice to complete their development. Eleven surviving pups were obtained. Nine were chimeras: six males and three females. Two of the males transmitted the embryonic stem cell genome containing the alteration in the hypoxanthine phosphoribosyltransferase gene to their offspring at high frequencies. These experiments demonstrate that a preplanned alteration in a chosen gene can be made in the germ line of an experimental animal by homologous recombination in an embryonic stem cell. PMID- 2573071 TI - Different classes of glutamate receptors mediate distinct behaviors in a single brainstem nucleus. AB - We have taken advantage of the increasing understanding of glutamate neuropharmacology to probe mechanisms of well-defined vertebrate behaviors. Here we report a set of experiments that suggests distinct roles for two major classes of glutamate receptors in a discrete premotor nucleus of the brainstem. The medullary pacemaker nucleus of weakly electric fish is an endogenous oscillator that controls the electric organ discharge (EOD). Its regular frequency of firing is modulated during several distinct behaviors. The pacemaker nucleus continues firing regularly when isolated in vitro, and modulatory behaviors can be reproduced by stimulating the descending input pathway. Glutamate agonists applied to the pacemaker in vitro produced increases in frequency, while glutamate antagonists selectively blocked stimulus-induced modulations. Experiments with glutamate antagonists in the intact animal resulted in specific effects on two well-characterized behaviors. Our data indicate that these behaviors are separately mediated in the pacemaker by receptors displaying characteristics of the kainate/quisqualate and N-methyl-D-aspartate subtypes of glutamate receptor, respectively. PMID- 2573072 TI - Grafting fibroblasts genetically modified to produce L-dopa in a rat model of Parkinson disease. AB - Rat fibroblasts were infected with a retroviral vector containing the cDNA for rat tyrosine hydroxylase [TH; tyrosine 3-monooxygenase; L-tyrosine, tetrahydropteridine:oxygen oxidoreductase (3-hydroxylating), EC 1.14.16.2]. A TH positive clone was identified by biochemical assay and immunohistochemical staining. When supplemented in vitro with pterin cofactors required for TH activity, these cells produced L-dopa and released it into the cell culture medium. Uninfected control cells and fibroblasts infected with the TH vector were grafted separately to the caudate of rats with unilateral 6-hydroxydopamine lesions of the nigrostriatal pathway. Only grafts containing TH-expressing fibroblasts were found to reduce rotational asymmetry. These results have general implications for the application of gene therapy to human neurological disease and specific implications for Parkinson disease. PMID- 2573073 TI - Dopamine synaptic complex with pyramidal neurons in primate cerebral cortex. AB - Dopamine (DA)-containing projections to the cerebral cortex are considered to play an important role in cognitive processes. Using a recently developed monoclonal antiserum directed against DA and an antibody directed against tyrosine hydroxylase in combination with Golgi impregnation and electron microscopy, we have observed that DA and tyrosine hydroxylase afferents establish symmetric membrane specializations with the soma, dendritic shafts, and spines of identified pyramidal cells in the prefrontal, cingulate, and motor cortex of primates. The axospinous contacts invariably formed part of a synaptic complex in which the dendritic spine of a pyramidal neuron was the target of both a DA positive symmetric and an unlabeled asymmetric bouton. This arrangement allows direct DA modulation of the overall excitability of cortical projection neurons by altering local spine responses to excitatory inputs. PMID- 2573075 TI - The effects of selective dopamine D1 or D2 receptor antagonists on the establishment of agonist-induced place conditioning in rats. AB - The ability of the dopamine D1 antagonist, SCH 23390 (0.01, 0.1, 1.0, 2.0 mg/kg) or the D2 antagonist, metoclopramide (1.0, 10.0, 20.0 mg/kg), to block the establishment of place conditioning with either the nonselective dopamine agonist, amphetamine (2.0 mg/kg), the D1 agonist, SKF 38393 (10.0 mg/kg), or the D2 agonist, quinpirole (1.0 mg/kg), was evaluated in rats. The experimental protocol consisted of three phases. During the preexposure phase, rats explored two distinctive compartments joined by a small tunnel. During the 8-day conditioning phase, rats were pretreated with either saline, SCH 23390 or metoclopramide; 1 hr later the animals were treated with an agonist and confined to one compartment for 30 min. On alternate days, rats received saline and were placed in the opposite compartment. Test days occurred over the remaining 3 days during which drug-free animals were allowed access to both compartments. A significant increase or decrease in the amount of time spent in the drug-paired environment was indicative of a place preference or aversion, respectively. SCH 23390 and metoclopramide were effective in blocking amphetamine-induced place preference and SKF 38393-induced place aversion. At lower doses, the D1 and D2 antagonist blocked the place preference induced by quinpirole, however, higher doses were not effective. In general, these data suggest that both receptor subtypes participate in the establishment of place conditioning with amphetamine, SKF 38393 or quinpirole. PMID- 2573074 TI - Nitric oxide mediates glutamate-linked enhancement of cGMP levels in the cerebellum. AB - Nitric oxide, which mediates influences of numerous neurotransmitters and modulators on vascular smooth muscle and leukocytes, can be formed in the brain from arginine by an enzymatic activity that stoichiometrically generates citrulline. We show that glutamate and related amino acids, such as N-methyl-D aspartate, markedly stimulate arginine--citrulline transformation in cerebellar slices stoichiometrically with enhancement of cGMP levels. N omega-monomethyl-L arginine blocks the augmentation both of citrulline and cGMP with identical potencies. Arginine competitively reverses both effects of N omega-monomethyl-L arginine with the same potencies. Hemoglobin, which complexes nitric oxide, prevents the stimulation by N-methyl-D-aspartate of cGMP levels, and superoxide dismutase, which elevates nitric oxide levels, increases cGMP formation. These data establish that nitric oxide mediates the stimulation by glutamate of cGMP formation. PMID- 2573077 TI - The effect of optical isomers of 3,4-methylenedioxymethamphetamine (MDMA) on stereotyped behavior in rats. AB - The relative potencies of S(+)-, R(-)-3,4-methylenedioxymethamphetamine (MDMA) and S(+)-methylene-dioxyamphetamine (MDA) in inducing stereotyped behavior were determined in comparison with p-chloroamphetamine. S(+)-MDMA was more potent than R(-)-MDMA in eliciting stereotyped behaviors such as sniffing, head-weaving, backpedalling and turning and wet-dog shakes. These results are consistent with the actions of the drug on release of neurotransmitters in which the S(+) enantiomer is more potent. The desmethyl derivative of (+)MDMA, (+)MDA, was more potent than (+)MDMA in eliciting stereotyped behaviors, and produced wet-dog shake behavior. PMID- 2573078 TI - Dopamine autoreceptor agonists attenuate spontaneous motor activity but not spontaneous fighting in individually-housed mice. AB - The present study was conducted to determine whether or not two behavioral characteristics of individually-housed mice, hyperactivity in a novel environment and intermale fighting, are attenuated by the dopamine (DA) agonists, apomorphine, (+)- and (-)-3-(3-hydroxyphenyl)-N-n-propylpiperidine (3-PPP). Autoreceptor-activating doses of these drugs which reduced spontaneous activity in a novel environment did not inhibit spontaneous fighting with conspecific olfactory bulbectomized males. Individually-housed mice were more active in a novel environment and showed a significant reduction of activity at lower doses of apomorphine, (+)- and (-)-3-PPP than group-housed mice. However, the ED50's for the inhibition of spontaneous activity in a novel environment in group- and individually-housed mice were similar: apomorphine, 0.02 vs. 0.012 mg/kg, SC; (+) 3-PPP, 0.50 vs. 0.51 mg/kg, SC; and (-)-3-PPP, 1.0 vs. 0.56 mg/kg, SC, for group- and individually-housed mice respectively. A significant proportion of individually-housed mice, but not group-housed mice, displayed catalepsy in response to high doses of (-)-3-PPP. These data suggest that DA autoreceptor agonists can modulate the hyperactivity syndrome but not spontaneous fighting behavior in individually-housed mice. PMID- 2573076 TI - Novel benzodiazepine receptor ligands: palatable food intake following zolpidem, CGS 17867A, or Ro23-0364, in the rat. AB - The potent imidazopyridine hypnotic, zolpidem, binds to central benzodiazepine receptors and has predominantly sedative properties, as determine in animal models. In tests of palatable food consumption in nondeprived male rats, the present results indicate that zolpidem (0.3-3.0 mg/kg) had no effect on food intake. Its lack of effect contrasts sharply with other benzodiazepine agonists which strongly stimulate palatable food intake. Two other novel compounds, both of which bind to benzodiazepine receptors, and which have reduced propensity to induce sedative effects, increased palatable food consumption, although in differing ways. The imidazobenzodiazepine Ro23-0364 (0.3-10.0 mg/kg) dose dependently increased feeding in the standard procedure, but failed to stimulate food intake in presatiated animals. The pyrazoloquinoline CGS 17867A (1.0-30.0 mg/kg) increased food intake in both test procedures, although the dose-effect relationship was nonmonotonic. Taken together, the data indicate a probable separation between hyperphagic and sedating effects of benzodiazepine receptor agonists. If zolpidem's sedative effect is linked to an action at a receptor subtype (benzodiazepine Type 1 or omega 1), then the hyperphagic effect of benzodiazepines may depend more on the alternative subtype. PMID- 2573079 TI - In vitro studies on the blood distribution of almitrine. AB - Blood binding of almitrine, a highly lipophilic drug, was investigated in vitro. [3H]-Almitrine was incubated in a serum pool and isolated protein and lipoprotein fractions. The investigations were performed by using ultracentrifugation and another method which measures the uptake by proteins from glass beads coated with almitrine. Our results with ultracentrifugation show that the distribution of almitrine in serum takes place predominantly in the lipoprotein fraction (78%) and to a minor extent (22%) in the fraction of d greater than 1.20 (albumin-rich fraction). Experiments using glass beads coated with almitrine were then conducted to measure the binding of almitrine to isolated plasma proteins. The maximal uptake values (mol almitrine/mol lipoprotein) of almitrine by isolated lipoproteins decrease from VLDL (260) to LDL (20) to HDL (3) and seem to be related to the lipid content of the particles. The uptake by albumin and alpha 1 acid glycoprotein was low. The molar ratios of [almitrine]/[lipoprotein] are roughly proportional to almitrine concentrations within the therapeutic range. When almitrine was incubated in erythrocytes suspended in several dilutions of serum, almitrine partitioned less in erythrocytes as the serum protein concentration increased in the suspension. PMID- 2573080 TI - Novel peripheral neurotransmitters and control of the airways. PMID- 2573081 TI - Biotechnology: Spinks eight years on. PMID- 2573082 TI - Biotechnology policy and achievement 1980-88. AB - Much progress has been made in line with the spirit of Spinks, but it is inevitable that perspectives have changed since 1980. At the research level, we know that much strategic work remains to be done before the full industrial benefits of biotechnology can be secured, even though there has been a gratifying improvement in academic-industrial liaison. The industrial progress and dissemination of biotechnology has been slower than Spinks implied, but we have a clear understanding of the importance of such contributors to the climate for investment as balanced regulation, training and public perception. The international dimension is important not only for the potential benefit of research and development programmes but also to monitor progress in the U.K. with respect to our competitors. Biotechnology now finds itself in a new framework of Government policy for innovation; there are new challenges in sustaining and adding to the impetus built up by the academic and industrial communities. PMID- 2573083 TI - Protein engineering and design. AB - Rapid advances in site-directed mutagenesis and total gene synthesis combined with new expression systems in prokaryotic and eukaryotic cells have provided the molecular biologist with tools for modification of existing proteins to improve catalytic activity, stability and selectivity, for construction of chimeric molecules and for synthesis of completely novel molecules that may be endowed with some useful activity. Such protein engineering can be seen as a cycle in which the structures of engineered molecules are studied by X-ray analysis and two-dimensional nuclear magnetic resonance. The results are used in the improvement of the design by using knowledge-based procedures that exploit facts, rules and observations about proteins of known three-dimensional structure. PMID- 2573084 TI - Genetic engineering applied to the development of vaccines. AB - The simplest application of the modern genetic manipulation methods to vaccine development is the expression in microbial cells of genes from pathogens that encode surface antigens capable of inducing neutralizing antibodies in the host of the pathogen involved. This procedure has been exploited successfully for development of a vaccine against hepatitis B virus (HBV) that is now widely used. Similar approaches have been directed towards formulations for immunization against several other animal and human diseases and some of these preparations are now presently in trials. Of no less importance is the impact of biotechnology in providing reagents for fundamental studies of topics such as the determination of virulence, antigenic variation, virus receptors and the immunological response to viral antigens. The core antigen of HBV is a good example of a product of genetic engineering that is a valuable diagnostic reagent, and that is finding important use in immunological studies of particular pertinence to vaccine development. PMID- 2573085 TI - Host-vector systems. AB - In 1980 it was only possible to express foreign genes in bacteria and a few easily cultured animal cells. During the subsequent eight years specialized vectors have been developed to allow the genetic manipulation of a wide range of both prokaryotes and eukaryotes. One of the major goals of biotechnology in 1980 was to use host cells as 'factories' for the production of proteins that were only available in minute quantities from natural sources. This has already lead to a new generation of pharmaceutical products. Advances in our understanding of host-vector systems have defined new goals. The basic concepts of expression vector design will be illustrated. Some of the new goals are discussed with particular reference to the exploitation of novel host-vector systems to develop vaccines and anti-viral agents against AIDS. PMID- 2573086 TI - Biosensors. AB - Biosensors are analytical devices that respond selectively to analytes in an appropriate sample and convert their concentration into an electrical signal via a combination of a biological recognition system and an electrochemical, optical or other transducer. Such devices will find application in medicine, agriculture, environmental monitoring and the bioprocessing industries. The last few years have seen great advances in the design of sensor architectures, the marriage of biological systems with monolithic silicon and optical technologies, the development of effective electron-transfer systems and the configuration of direct immunosensors. Recent progress in these areas has already led to the introduction of new-generation biosensors into the competitive diagnostics market place. PMID- 2573087 TI - Biochemical engineering challenges of purifying useful proteins. AB - Some of the biochemical engineering issues that arise when the purification of proteins is scaled up are examined. The first question addressed is the quantity of various proteins likely to be required. Consideration is given to the order in which isolation procedures may be applied and to the selective removal on a large scale of non-proteinaceous materials. Some general problems such as the effect of mechanical forces on proteins and their complexes and aggregates are examined, together with ways of enhancing the properties of these aggregates and the methods of recovering them. The manner in which on-line process optimization can be achieved during pilot plant trials is discussed and the paper concludes with a summary of new scientific developments to which the biochemical engineer will need to respond. PMID- 2573088 TI - The management and coordination of biotechnology in the U.K. 1980-88. AB - Government policy towards biotechnology has come a long way since the Spinks Report. Spinks advocated centralized coordination of policy, an approach deliberately rejected in 1981 by the Government in favour of continued pluralism, with each of the scientific research councils and various ministries 'doing their own thing'. This has led to considerable diversity of activity, and during these eight years more has in fact been achieved than is often recognized. But it also created an overlapping of responsibilities with concomitant friction and bad feeling that has wasted time and resources. The paper argues that some degree of friction is inevitable. By their nature new technologies cut across existing disciplines and blur institutional boundaries. The traditional approach has been to muddle through, allowing new institutions to emerge and adapting the old as seems appropriate. Lack of resources, however, argues against too brash a competitive approach. The paper suggests that strategic or precompetitive research should be seen as a complement to, rather than competitive with basic research, and cautions against too radical a restructuring of institutions at the present time. PMID- 2573089 TI - Antibody engineering. AB - The antibody molecule is a therapeutic agent, designed by nature to bind to a wide range of antigen molecules and to trigger effector functions, such as complement lysis and cell-mediated killing. The genes encoding antibodies can be manipulated in vitro, allowing the binding sites for antigen and effector molecules to be dissected, and new properties to be engineered. The future for the application of engineered antibodies in medicine is reviewed in the context of the past century. PMID- 2573090 TI - Antibiotics: opportunities for genetic manipulation. AB - New antibiotics can still be discovered by the development of novel screening procedures. Notable successes over the last few years include the monobactams, beta-lactamase inhibitors (clavulanic acid) and new glycopeptides in the antibacterial field; antiparasitic agents such as avermectins; and herbicidal antibiotics like bialaphos. In the future we can expect the engineering of genes from 'difficult' pathogens, including mycobacteria and fungi, and cancer cells, to provide increasingly useful in vitro targets for the screening of antibiotics that can kill pathogens and tumours. There will also be a greater awareness of the need to reveal the full potential for antibiotic production on the part of microorganisms by the physiological and/or genetic awakening of 'silent' genes. Nevertheless, the supply of natural antibiotics for direct use or chemical modification is not infinite and there will be increasing scope for widening the range of available antibiotics by genetic engineering. 'Hybrid' antibiotics have been shown to be generated by the transfer of genes on suitable vectors between strains producing chemically related compounds. More exciting is the possibility of generating novelty by the genetic engineering of the synthases that determine the basic structure of antibiotics belonging to such classes as the beta-lactams and polyketides. Research in this area will certainly yield knowledge of considerable scientific interest and probably also of potential applicability. In the improvement of antibiotic titre in actinomycetes, protoplast fusion between divergent selection lines has taken a place alongside random mutation and screening. In some cases the cloning of genes controlling metabolic 'bottlenecks' in fungi and actinomycetes will give an immediate benefit in the conversion of accumulated biosynthetic intermediates to the desired end product. However, the main impact of genetic engineering in titre improvement will probably come only after a further use of this technology to understand and manipulate the regulation of antibiotic biosynthesis as a facet of the general challenge of understanding differential gene expression. Streptomyces offers a particularly fertile field for such research, following the isolation of DNA segments that carry groups of closely linked operons for the biosynthesis of and resistance to particular antibiotics, and of genes with pleiotropic effects on morphological differentiation and secondary metabolite formation. PMID- 2573091 TI - Animal biotechnology. AB - Biotechnology has taken two directions in efforts to speed up animal production above the rates achievable by selective breeding. Recombinant DNA methods have been used to engineer protein gene products for direct administration to livestock, as in recombinant growth hormone to stimulate lactation in dairy cows or yield faster-growing, leaner carcasses in meat animals. Cloned cellulolytic genes have been inserted into ruminal microorganisms with a view to improving ruminant nutrition. The other direction is to use advanced breeding technologies to enhance performance. These include laboratory culture of large numbers of viable embryos for non-surgical transfer to surrogate mothers, development of methods for sexing sperm and embryos, cloning embryos by nuclear transplantation and gene transfer to create livestock with superior performance traits. In all cases material progress will depend upon a deeper understanding of the underlying physiological and developmental control mechanisms and public confidence that due regard is being paid to animal welfare, and to social and environmental implications. PMID- 2573092 TI - Exploitation of biotechnology in a smaller company. AB - The establishment of a research-orientated biotechnology company was one of the recommendations of the Spinks Committee. How the National Enterprise Board, the Medical Research Council and private sector financial institutions responded to this recommendation, leading to the formation of Celltech in November 1980, is described. The paper then outlines Celltech's current position (it is no longer a small company) and future plans and describes the two areas of technology in which the company has achieved a leading position: mammalian cell culture and the engineering of the genes coding for monoclonal antibodies. Celltech's methods of working with academia and the benefits to both are then described, and finally the paper tries to reach some conclusions about the part smaller companies play in technological innovation. PMID- 2573093 TI - Exploitation of biotechnology in a large company. AB - Almost from the outset, most large companies saw the 'new biotechnology' not as a new business but as a set of very powerful techniques that, in time, would radically improve the understanding of biological systems. This new knowledge was generally seen by them as enhancing the process of invention and not as a substitute for tried and tested ways of meeting clearly identified targets. As the knowledge base grows, so the big-company response to biotechnology becomes more positive. Within ICI, biotechnology is now integrated into five bio businesses (Pharmaceuticals, Agrochemicals, Seeds, Diagnostics and Biological Products). Within the Central Toxicology Laboratory it also contributes to the understanding of the mechanisms of toxic action of chemicals as part of assessing risk. ICI has entered two of these businesses (Seeds and Diagnostics) because it sees biotechnology making a major contribution to the profitability of each. PMID- 2573095 TI - Yawning induced by the selective dopamine D2 agonist N-0437 is blocked by the selective dopamine autoreceptor antagonist (+)-UH 232. AB - Yawning and stretching responses were elicited in rats by a small dose (0.3 mg/kg) of the highly selective dopamine D2 agonist, N-0437. The responses were blocked by the highly selective dopamine autoreceptor antagonist, (+)-UH 232 (3.0 mg/kg), but not by raclopride at a dose which selectively blocks postsynaptic D2 receptors. The results strongly confirm the view that yawning and stretching are behavioral responses elicited by stimulation of presynaptic D2 receptors. PMID- 2573094 TI - The effect of PGE in multiple experimental models. II. Effect on steady-state levels of plasma and brain amino acids and transmitters. AB - Elevations in prostaglandin E (PGE) have been documented in tumor and trauma patients. The physiologic significance of this elevation is not fully established. Utilizing a long-acting derivative of PGE, 16,16-dimethyl prostaglandin E (dPGE), in a rat model, we evaluated its effects on metabolic rates, amino acid, and neurotransmitter metabolism. dPGE was not found to significantly alter resting metabolic rate. It did decrease the plasma level of three amino acids, including tyrosine. Although dPGE also decreased brain tissue levels of tyrosine, no significant alterations were observed on amine neurotransmitters or metabolites. PMID- 2573096 TI - Effects of prenatal alcohol on gamma-glutamyl transpeptidase in various brain regions. AB - Prenatal exposure of rats to alcohol produces morphological, biochemical, behavioral and physiological abnormalities. The enzyme gamma-glutamyl transpeptidase activity is increased when an animal is exposed to alcohol chronically. We examined the effect of the in utero exposure to alcohol on the regional brain distribution of gamma-GTP. Pregnant rats were placed into the following treatment groups: LC (ad lib lab chow and water), PF (pair-fed), 10% EDC (ethanol derived calories), 20% EDC and 35% EDC. The LC group was fed lab chow and water ad lib, the PF, 10% EDC, 20% EDC and 35% EDC groups were fed a liquid diet containing either 0%, 2%, 4% or 6.7% v/v ethanol, respectively. All the treatment groups received the same volume of an isocaloric diet as was consumed by the 35% EDC group. At birth, litters were culled to six and cross fostered to untreated surrogate mothers. Pups were sacrificed at 30 days of age and brains dissected into 8 regions. Each brain region was homogenized and divided into soluble and membrane bound fractions by centrifugation and digestion with deoxycholic acid. gamma-GTP activity was then measured. It was determined that the in utero exposure to alcohol produces an increase in brain gamma-GTP activity. The increase in gamma-GTP activity in some areas of brain is maintained at least until the animals are 30 days old. Alcohol treatment had no effect on the activity associated with the soluble form of enzyme. However, the activity associated with the membrane bound form of the enzyme was increased in several brain regions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573097 TI - Substitution of temazepam and midazolam in pentobarbital-dependent rats. AB - The ability of temazepam and midazolam to substitute for pentobarbital and thus maintain the physical dependence state was used to assess the potential dependence liability of these two benzodiazepine compounds. Male Sprague-Dawley rats, weighing 175-200 g and having ad lib access to food and water, were determined to be dependent on pentobarbital following 12 days of continuous, intraperitoneal infusion of pentobarbital using an escalating drug infusion schedule. On day 13 (substitution phase) the pentobarbital was replaced with either temazepam, midazolam or vehicle and the rats were infused for an additional 24 hours. This was followed on Day 14 (withdrawal phase) by a 24 hr saline infusion period. Rats were observed for changes in overt behavior and alterations of body weight during both Day 13 and Day 14. Preliminary potency estimation studies had indicated that both drugs were more potent and longer acting than was pentobarbital. Temazepam, in doses of 32.5, 65 and 130 mg/kg/24 hr, was demonstrated to substitute for pentobarbital and provided dose-dependent suppression of overt behavioral signs indicative of withdrawal. Temazepam also suppressed the weight loss typically observed during withdrawal. Substitution of saline for temazepam resulted in an increased incidence of withdrawal signs and an approximate 10% decline in body weight. Midazolam, in doses of 60 and 120 mg/kg/24 hr, also substituted for pentobarbital and suppressed both overt behavior and weight loss. Following saline substitution on Day 14, a mild withdrawal syndrome was evident although body weight was noted to remain near control values.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573099 TI - [Neuroleptic drugs in geriatric psychiatry]. PMID- 2573098 TI - [Comparative studies of the effects of perazine, fluphenazine, trifluoperazine, chlorpromazine and haloperidol on primary and deficit symptoms of paranoid schizophrenia]. AB - In order to evaluate the influence of some neuroleptics on unproductive symptoms of schizophrenic process, first hospitalized patients with paranoid schizophrenia underwent monotherapy with perazine, fluphenazine, trifluoroperazine, chlorpromazine, and haloperidol. All the drugs eliminated the productive activity of the disease process within 2 months in both women and men aged 18-40 years. However, drugs showed different qualitative and quantitative effects on unproductive symptoms of schizophrenic process; this fact allows the suggestion regarding the particular usefulness of drugs in the out-patient care. PMID- 2573100 TI - [Clinical aspects, pathogenesis and treatment of neuroleptic malignant syndrome]. PMID- 2573101 TI - [Dopanergic agonists and schizophrenia]. AB - The neutrotransmitter dopamine mediates its central nervous system actions via 2 types of receptors: D1 and D2, each of which exist in a low or a high affinity state. In schizophrenic patients, it appears that part of the symptomatology may be secondary to dopaminergic hyperactivity (the so called positive symptoms), whereas negative symptoms would be secondary to structural damage. Antipsychotic drugs that are used in clinics are antidopaminergic. It is interesting to note that those drugs are ineffective in part of the schizophrenic population, the chronic one. This inefficiency might be due to primary resistance or to adaptation of the central nervous system to the drugs. Some authors tried dopamine agonists to treat schizophrenic patients. Apomorphine, N-n propylnorapomorphine, l-DOPA and bromocriptine were tried. The purpose of these substances was to desensitize dopaminergic system, or to inhibit it via presynaptic autoreceptor stimulation, depending of the study. Some authors hypothesized a drop in frontal cortical dopaminergic system to explain part of the schizophrenic symptomatology. Dopamine agonists might act at this level to enhance frontal dopaminergic activity. After those experimentations with dopamine agonist drugs, it appears that 2 of them might have efficiency and clinical use: l-DOPA and bromocriptine. PMID- 2573102 TI - Immune dysfunctions in psychiatric patients. AB - Alterations in the serum proteins and immunoglobulins have been investigated in the search for the possible etiological causes of schizophrenic patients. The results, so far, have been varied and inconclusive. We assayed serum immunoglobulins, and antinuclear antibody (ANA) titres in 94 randomly selected, recently hospitalized, psychotic patients. A complete physical, neurological, laboratory and electrophysiological examination was conducted on each patient. Systems survey and family, as well as past histories were obtained. Our results indicated that 12 patients had high titres of ANA and none of them had any other clinical evidence of autoimmune disease. This finding did not correlated with age, sex, diagnosis, or duration if illness, but an association with the medications that the patients were receiving, was noted. The elevation in ANA titres seems to be related to use of lithium as well as neuroleptics. Our data suggests that all neuroleptics can produce high ANA titres. Systematic study of this phenomenon and their long-term consequences warrant further exploration. PMID- 2573103 TI - Agranulocytosis and significant leucopenia with neuroleptic drugs: results from the AMUP program. AB - In the Federal Republic of Germany adverse drug reactions (ADR) have been continuously assessed at the departments of Psychiatry of Berlin and Munich since May 1979. About 13,000 neuroleptic-treated inpatients were monitored until August 1988. Approximately 1100 patients were exposed to clozapine, 6800 to haloperidol and 6000 to perazine, the two most frequently used neuroleptic drugs. In this 9 year period seven cases of agranulocytosis were observed, all in women. One case occurred with clozapine in monotherapy, the other six with perazine, three times in monotherapy, once in combination with trimethoprim/sulfamethoxazole and in one case each in combination with tricyclic antidepressants. Significant leucopenia (less than or equal to 3000/mm3) was observed in an additional eight cases. On four occasions each butyrophenones (twice in combination with TCA) and tricyclic neuroleptics (once in combination with TMS) were involved. The number of exposed patients per drug is too small for calculation of statistically valid incidence rates, especially in view of the frequent polypharmacy. The course of agranulocytosis was benign in all seven cases and required no other treatment than drug withdrawal in three cases. The early detection by regular WBCs is supposed to be mainly responsible for this and is therefore recommended at weekly intervals. This measure of safety appears most important for all medium potency tricyclic neuroleptics. As to treatment of agranulocytosis, additional measures (antibiotics, intensive medical care) depend upon the severity of the clinical picture. PMID- 2573104 TI - D1- and D2-dopamine receptor occupancy during treatment with conventional and atypical neuroleptics. AB - Using positron emission tomography and the selective ligands 11C-SCH23390 and 11C raclopride, central D1- and D2-dopamine receptor occupancy was determined in schizophrenic patients treated with clinical doses of classical and atypical neuroleptics. Treatment with ten chemically distinct classical neuroleptics resulted in a 65-89% occupancy of D2-dopamine receptors. This finding represents strong support for the hypothesis that the mechanism of action of antipsychotic drugs is indeed related to a substantial degree of D2-dopamine receptor occupancy. In two patients treated with the atypical neuroleptic clozapine, 300 mg b.i.d. and 150 mg b.i.d., the D2-dopamine receptor occupancy was 65 and 40%, respectively. D1-dopamine receptor occupancy was determined in six antipsychotic drug-treated patients. No D1-dopamine receptor occupancy was found in patients treated with sulpiride and perphenazine, compounds known to be selective D2 dopamine receptor antagonists. The highest D1-dopamine receptor occupancy, 42%, was found in the patient treated with clozapine 150 mg b.i.d. The effects of the atypical neuroleptic clozapine may be related to a combined effect on both D1- and D2-dopamine receptors. PMID- 2573105 TI - Effect of neuroleptics on positive and negative symptoms and the deficit state. AB - The concept of negative symptoms tries to operationalize a deficit syndrome observed in schizophrenia, but also in other disorders. The instruments for the measurement developed so far are unclear in their dimensional structure and validity. Further methodological development is needed. A new scale for measuring negative symptoms was derived from the AMDP-system and applied to results of drug trials with clozapine, fluperlapine, and haloperidol. The three drugs were equally effective on negative symptoms of acute and chronic schizophrenics. PMID- 2573106 TI - Biochemical and behavioural properties of clozapine. AB - The selection and early development of clozapine was based upon its gross behavioural, arousal-inhibiting, sleep-promoting, and caudate spindle-prolonging properties. Compared to classical neuroleptics, clozapine causes only a short lasting elevation of plasma prolactin levels, elevates both striatal homovanillic acid and dopamine content, is devoid of marked apomorphine-inhibitory or cataleptogenic activity and fails to induce supersensitivity of striatal dopaminergic systems after chronic administration. Clozapine's intrinsic anticholinergic activity, while stronger than that of other neuroleptic agents, does not appear to underlie either its failure to induce tardive dyskinesias or its superior antipsychotic activity. Furthermore, the overlap between clozapine and several classical neuroleptics with regard to alpha-adrenergic-, serotonin- and histamine-blocking activity makes it unlikely that one or more of these properties is the key to its atypical characteristics. More recent findings show that clozapine and classical neuroleptics differ with regard to their indirect effects on nigral GABA-ergic mechanisms implicated in the induction of tardive dyskinesias and, possibly in keeping with this, that clozapine and similar agents exhibit preferential blockade of D-1 dopamine receptors in the whole animal. Such an action of clozapine in man could well explain both its low EPS liability and, in some subjects, its superior antipsychotic activity. PMID- 2573108 TI - Louis Agassiz and the discovery of the coelenterate nervous system. PMID- 2573109 TI - [Oral premedication with Benzodiazepine]. PMID- 2573107 TI - Clinical experience with clozapine in Germany. AB - The so-far successful development of the atypical neuroleptic clozapine had been interrupted by the Finnish epidemic of agranulocytosis in 1975. However, though the strong regulatory control of prescribing clozapine, since then its clinical use has increased steadily, particularly recently. This may be due mainly to the considerable number of patients whose psychotic states or at least whose negative symptoms do not respond to other neuroleptics, or who have problems with extrapyramidal side effects. Due to its significant antipsychotic, and to its probable even if mild antidepressive efficacy as well as to its possible efficacy against some negative symptoms of schizophrenia, clozapine is currently a real and indispensable alternative to other existing neuroleptics. Further research should be directed both to the clinical validation of the latter mentioned therapeutically desired effects and to the causes and predictability of agranulocytosis, as yet the main risk of clozapine therapy. Theoretically profitable would be the clarification of the causes of hyperthermia and particularly of those of the mutual independency of extrapyramidal motor disturbances and antipsychotic efficacy. PMID- 2573110 TI - The pharmacology of segmental transmission in the spinal cord. PMID- 2573111 TI - Neuropharmacological and physiological validation of a computer-controlled two compartment black and white box for the assessment of anxiety. AB - 1. The two-compartment black and white box first described by Crawley and Goodwin (1980) has been used to study anti-anxiety properties of drugs but has not been validated. 2. An automated test system and validation of the protocol for the evaluation of compounds with anxiolytic or anxiogenic potential is described. 3. The box is partitioned into black and white sections with an interconnecting opening and is equipped with micro-switch photoelectric controls (light source and photoreceiver) and an interface connected to the menu-driven computer during anxiety testing. 4. Plasma corticosterone levels in naive mice maintained on a reversed L:D cycle was significantly reduced following restricted exposure to the brightly lit white section but not in the red-illuminated black section. 5. The optimal structural configuration in different test situations was found to be a square rather than a round box. 6. Under normal conditions, mice spend about 60% of the time in the dark compartment so that the exploratory activities and time spent in the white section are taken as a measure of anxiety. 7. Compounds examined included the reference anxiolytic diazepam, nicotine, naloxone, MDL 72222, ICS 205 930 and buspirone, all of which increased mouse exploratory activities in the white section. PTZ, beta-CCP, morphine and amphetamine increased exploration in the black compartment and reduced exploration in the white area. 8. Fluphenazine and imipramine had no specific effects on anxiety responding, although the cataleptogenic effect of fluphenazine was apparent. 9. Daily repeated testing was possible with a maximum of up to four trials a week using naive animals during the 5-min test session. 10. The results suggest that the rapid and automated test system for the assessment of changes in measures of anxiety is not only valid for large scale evaluation of compounds but could be used to elucidate mechanisms of drug action and the CNS pathways linked with anxiolysis and/or anxiogenesis. PMID- 2573112 TI - [Phencarol in the treatment of allergic dermatoses of various pathological mechanisms]. AB - Phencarol produced in the USSR is an antihistaminic drug highly effective in allergic dermatoses with the pathological mechanism of humoral and cellular type. The best results were obtained in acute allergic dermatoses and in acute recurrences of chronic allergic diseases. In comparative studies with difergan a greater efficiency of phencarol was noted, which was expressed as a higher proportion regression of lesions and more rapid action of the drug. The duration of phencarol treatment was about 3 weeks, the daily dose was 50-75 mg in adults. The drug was well tolerated. PMID- 2573113 TI - [Use of first and second generation antihistaminics in pediatric patients with allergic rhinitis]. AB - The authors studied 75 outpatients seen in the Pediatric Allergy Clinic of the Social Security Metropolitan Hospital Complex and of the Paitilla Medical Clinics, between 4 and 14 years of age, 28 females and 47 males, because they were suffering from allergic rhinitis and most of them (84%) had an elevated immunoglobulin E level. Skin tests were positive with house dust (100% of patients), house dust mites (95%), molds (92%) and grasses (22%). Symptoms occurred at any time of the year in 56 (73%) patients, and in the rainy season in only 20 (27%). All were examined by the same investigator at the beginning of the study and at the end of the first and second week. A family member evaluated the symptoms daily in the morning and at night. For this purpose the following scale was used: 0 = no symptoms; 1 = mild; 2 = moderate; 3 = severe symptoms. The symptoms evaluated were nasal congestion and sneezing, rhinorrhea and conjunctival congestion, post nasal drip and pharyngeal irritation. The signs that were evaluated were dark circles around the eyes, hypertrophy of the turbinates, the color of the nasal mucosa, the presence of polyps and deviation of the nasal septum. This study showed that Terfenadine improved the rhinitis in 87.5% of the patients, that Clemastine was effective in 70.5%, and that placebo, in 43%. Also, somnolence was observed in 4% and dryness, in 43% of patients using Terfenadine, while Clemastine induced somnolence in 32% of patients. PMID- 2573114 TI - [Use of somatostatin in the conservative treatment of external pancreatic fistula]. AB - We present six cases of postoperative pancreatic fistulas treated with complete diet and total parenteral nutrition. Due to the slow closure of the fistula, we added somatostatin to treatment (doses specified in the report). Flow disappeared completely in all cases, and the fistula was successfully closed in 5 of the 6 cases. There were no side effects due to the medication. We emphasize that the results of somatostatin treatment were favorable, achieving early closure of the fistula and reducing the complications derived from it. PMID- 2573115 TI - [Pharmacologic therapy of bronchial asthma]. AB - The main feature in asthma is bronchial eosinophilic inflammation, induced, in genetically predisposed subjects, by known (aeroallergens, viral infection, occupational agents) and unknown agents. Inflammation critically increases bronchial responsiveness so that even rather mild stimuli (exercise, cold air, mist, allergens) are able to induce obstructive reactions. When preventive measures (specific immunotherapy, avoidance of inflammatory and precipitating factors) are inadequate, a pharmacological treatment is necessary. This should, in addition relieve bronchospasm, neutralize precipitating factors and reduce inflammation. In the first case bronchospasmolytic antireactive agents (beta 2 stimulants, antimuscarinic agents, theophyllines) should be used, whereas, in the second case, the first choice drugs are corticosteroids. Chromon derivatives can also be considered for both their antireactive and antiflogistic effects. It is impossible to elaborate rigid therapeutic schemes. Therapy must be individualized on the basis of symptoms, objective physical signs and functional data. Drugs, except theophylline (for which slow release oral formulations are preferred), should be preferably administered by inhalation. For patients who cannot properly master the use of metered dose inhalers, spacer devices or dry powder inhalers are indicated. Whereas mild asthmatic forms can be treated without corticosteroids, these are absolutely necessary when bronchospasmolytics and chromon derivatives, although appropriately used, do not provide a satisfactory control of the asthmatic syndrome. Many other drugs are now under study (antileukotrienes, anti-PAF, anti-5-lipoxygenase, anti-phospholipase A2), but the results published so far do not seem very promising. Anti-histamines, calcium antagonists, alpha-blocking drugs are justified only in some circumstances. The antireactive activity of loop diuretics is interesting, but its real therapeutic value is still to be assessed. Antibiotics are only needed in the (rare) cases of worsening of asthmatic symptoms due to infective sensitive agents. PMID- 2573116 TI - [Medical therapy of allergic rhinitis]. AB - In the last few years, owing to new pathophysiological and pharmacological notions, the therapy of allergic rhinitis has achieved some remarkable goals. On the basis of personal experience and of the literature, a close examination of drugs used for aspecific treatment of allergic rhinitis is carried out, mainly regarding the most currently useful and effective. PMID- 2573117 TI - [Pharmacologic control of breathing]. AB - Recent advances in respiratory neuropharmacology and neurophysiology have allowed the assessment of the effects of different drugs on the control of the ventilation both qualitatively (alterations of ventilatory pattern) and quantitatively (size and duration of the ventilatory and haemogasometric alterations). In the control mechanism of ventilation, the pharmacological intervention can act both on the respiratory input (basal metabolism, peripheral chemo-receptors, pulmonary receptors, bulbar neurons, cortical nervous system) and on the respiratory output (respiratory muscles). Based on personal experience in this field and the recent literature, the Authors briefly discuss the seat and the mechanism of action of the drugs with stimulating effect (respiratory analeptics, almitrines, progesterone, acetazolamide, salicylates, protriptyline, theophylline) and depressing effect (narcotics and narcotic-antagonists, anaesthetics, barbiturates and benzodiazepines) on the ventilation, as well as the role of the neurotransmitters and modulators. The clinical (positive and negative) effects of these drugs, particularly related to the patients with chronic lung disease, are also illustrated. PMID- 2573118 TI - Restriction fragment length polymorphism of the human insulin receptor gene among Mexican Americans. AB - Recently, we examined a restriction fragment length polymorphism (RFLP) at the insulin receptor locus in Mexican Americans. This RFLP can be detected using the restriction endonuclease Rsa I, and has three alleles of 3.4 kb, 6.2 kb, and 6.7 kb. Our data suggested that the 3.4 kilobase pair (kb) allele may be associated with type II (non-insulin dependent) diabetes mellitus in Mexican Americans. We initiated studies to identify additional polymorphisms at the insulin receptor locus that might be useful in further studies on the association of type II diabetes and the insulin receptor gene. During the course of these studies we observed that the 6.7 kb and 6.2 kb alleles of the RsaI polymorphism appears to be due to an insertion or deletion of DNA sequences, so that DNA fragments of different lengths are generated when DNA from heterozygous individuals is digested with selected restriction endonucleases that cut on either side of this region. The 3.4 kb allele is apparently due to a site specific polymorphism. In the present report, results of these findings are presented as well as a description of additional polymorphisms that we have identified among Mexican Americans. PMID- 2573119 TI - Modulation of gamma-glutamyl transpeptidase activity by dietary eicosapentaenoic acid and vitamin E in livers of rabbits on hypercholesterolemic diet. AB - Rabbits were fed high-cholesterol diets containing either eicosapentaenoic acid (EPA) or vitamin E at doses of 80 mg and 100 IU per day, respectively. Liver gamma-glutamyl transpeptidase (GGT) activity and liver cholesterol and phospholipid levels were determined following the administration of the diets for 45 days. The feeding of cholesterol produced the highest concentrations of cholesterol in livers accompanied with the elevated enzymatic activity. Addition of EPA to the diet dramatically reduced GGT activity to normal levels, whereas vitamin E administration caused only a slight reduction. PMID- 2573120 TI - Ciclosporin and thyroid-stimulating immunoglobulins in endocrine orbitopathy. AB - The study investigated whether ciclosporin (C) affected the thyroid-stimulating immunoglobulins (TSI) in serum of patients with endocrine orbitopathy (EO). The effect of C was compared with that of prednisone (P). Fifteen patients with EO classes III-V received C (n = 7) or P (n = 8). In addition to the immunosuppressants, five patients with Graves' disease in each group received methimazole (MMI). The stimulation of the cAMP levels in the medium of thyrocyte cultures was determined as a parameter of TSI. The TSI levels were markedly lowered in both groups during and after therapy. C group: before therapy 6.2 pmol/ml +/- 1.63 (100%, mean +/- SEM), during treatment 4.6 pmol/ml +/- 2.28 (74%), after treatment 4.1 pmol/ml +/- 1.33 (66%). P group: before treatment 9.1 pmol/ml +/- 3.42 (100%), during treatment 5.9 pmol/ml +/- 2.90 (65%), after treatment 3.7 pmol/ml +/- 1.20 (41%). There is neither a significant difference between the two groups nor between the patients who received the combined therapy (MMI + immunosuppressants) or only received immunosuppressants (P more than 0.05). The mean cAMP value of the healthy reference group (n = 19) is 0.4 pmol/ml +/- 0.03. There is a significant difference between this value and the cAMP values of the patients both before and after therapy (P less than 0.01). Thus, both C and P markedly lower the TSI titers of patients with EO. PMID- 2573121 TI - [Current developments in the pharmacology of dopaminergic receptors (I)]. PMID- 2573122 TI - [The physiopathology of the basal ganglia and the etiopathogenesis of Parkinson's diseases]. PMID- 2573123 TI - [Current developments in the pharmacology of dopaminergic receptors (II)]. PMID- 2573124 TI - [Controversies and view points in the problem of ischemic cardiopathy]. AB - The paper reports both on the new directions and trends and on the present stage of several controversial problems, for a better understanding of etiopathogenesis prevention and treatment of ischemic cardiopathy, a priority problem of the public health in Romania. PMID- 2573125 TI - [Proarrhythmias and anti-arrhythmic effects in the prolonged repolarization syndrome]. PMID- 2573126 TI - [Coronary spasm and its current status]. PMID- 2573127 TI - [Echocardiographic parameters in the follow-up of left ventricular performance before and after the surgical correction of mitral and aortic regurgitation]. AB - The present study investigates 16 patients with aortic regurgitation (AR), and 10 patients with mitral regurgitation (MR). Before operation, all the 26 patients had increased telediastolic and telesystolic diameters. The patients were divided into two groups, as a function of the postoperative values of the telediastolic diameter: group A--with the telediastolic diameter within normal limits; group B- with the persistence of the increased values of the telediastolic diameter. In the case of the patients with AR, the following postoperative situation was noticed: in group A: the peak systolic stress (PSS) was increased, the shortening fraction (SF) in the telesystolic stress (TSS) within normal limits; in group B: PSS and TSS were increased, and ST depressed. 8 months after the valvular correction, in group A: PSS, TSS and SF were normal; in group B: the changes in PSS and TSS continued and was lowered. In the case of the patients with MR, before operation, in group A: PSS was slightly increased while TSS and SF were normal; in group B: PSS was slightly increased, TSS, increased and SF depressed. 8 months after the surgical correction: in group A: PSS and SF decreased; in group B: echocardiographic changes continued. Therefore, in the group B patients with MR and AR, the persistence of the increased values of PSS, TSS leads to the postoperative decrease of SF (post-charge in excess). PMID- 2573129 TI - [The therapeutic implications of biostimulation using an athermic laser in rheumatology]. PMID- 2573128 TI - [Nifedipine (corinfar) in the treatment of arterial hypertension (AHT)]. AB - The paper studies the effect of a blocking agent, calcium-niphedipine (corinfar), single or associated with other hypertensors (diuretic, sympathico-colitic), in a dose of 30 mm (3 X 10 mg) administered for about 8-10 days on a lot of 90 patients, men and women, with essentially noncomplicated AHT or associated with: pyelonephritis, cardiac insufficiency, ischemic cerebral stroke, rhythm and conduction disturbances. The systolic AT values of the patients ranged between 160 and 300 mmHg and the diastolic AT between 90 and 150 mmHg. The systolic AT decreased significantly with 40-117 mmHg and the diastolic AT with 15-55 mmHg. The drug was well tolerated and no side effect was noticed. In two cases, the AT values were not influenced by the treatment. PMID- 2573130 TI - [Sources of error in the diagnosis of pleurisy]. PMID- 2573131 TI - [Observations on 35 cases of strongyloidiasis hospitalized at a clinical digestive disease unit]. AB - After a short presentation of the etiopathogenesis of this parasitosis, the authors report their observations on 35 cases of strongyloidiasis followed in the recent years. On the basis of the epidemiologic and clinical analyses of these cases, the authors show that some data, i.e. those on the environment to which the patients belong, or their occupation, and also some clinical manifestations (clinical polymorphism) may suggest the diagnosis of strongyloidiasis in the immunosuppressed in about 30% of the cases. An associated blood eosinophilia may be a more possible for the diagnosis of strongyloidiasis. The positive diagnosis has to take into consideration, beside the repeated diet exam, the method of cultures and also jejunal biopsies, which may also verify the efficiency of the treatment recommended. The authors recommend the treatment with Mintezol and, with lower results, Vermigal and Mebendazol. PMID- 2573132 TI - [The role of atopic background in the genesis of allergic manifestations in intestinal parasitoses]. AB - The previous literature and personal observations showed the predominant role of the atopic background in the appearance of the allergic manifestation in the parasitic infestation and especially in giardiasis. The lot investigated was made of 52 patients with allergic manifestations, also infested with Giardia intestinalis or with Ascaris lumbricoides. The hereditary collateral and personal allergic antecedents and the clearly deficient histaminopexy (1/20) were present in 1/2 of the patients. Summation of the various parameters of atopic background shows its presence in about 2/3 of the patients investigated. Association of the clinical involvement of trophallergens was present in 1/2 of the patients. The results show that the allergic manifestation in the intestinal parasitoses have a multifactorial determination. The atopic background is predominant in their development. These manifestations are often a result of alimentary allergy due to the absorption of alimentary antigens through the enteral mucous membrane affected by the parasite. PMID- 2573133 TI - [The effect of a plant mixture on the metabolic equilibrium in patients with type 2 diabetes mellitus]. AB - The present paper analyses the results obtained in 82 patients with diabetes mellitus of the 2nd type: 59 women and 23 men, between 41 and 74 years old (average +/- DS, 58 +/- 9 years), of which 58 had an index of the body weight higher than 26. The diabetes duration ranged between newly discovered and 11 years. Each patient was given, 3 times a day, a 150 ml cup containing an infusion of the following mixture of plants previously cut into small pieces: Phaseolus vulgaris (pod), Morus alba (leaf), and Vaccinum myrtillus (leaves). The approximate dose used was of about 15 g/day. The treatment lasted for two months. Before and after treatment, the following parameters were determined: Hb Al (Bio Rex method) in 31 cases; the average of 3 consequent glycemias; the value of glycemia and insulinemia recorded after a standard lunch, consisting of about 40 g glucides, 14 g proteins and 6 g lipids (50 g bread, a boiled egg and a boiled apple of 100 g). Analysis of the results obtained enabled the following temporary conclusions (1). In 74 out of the 82 cases studied, the average values of glycemia, after the treatment with plants, were lower than those recorded before the treatment (the average values of the whole lot: 219 +/- 82 mg/dl before treatment and 166 +/- 76 mg/dl after treatment (2). The overall decrease recorded, of 53 mg/dl, represents 24.3% of the initial value (3).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573134 TI - [The "stiff heart" syndrome]. PMID- 2573136 TI - [Treatment of peptic disorders: are all drugs permitted?]. PMID- 2573135 TI - [Recent concepts on the innervation of the lower urinary tract and its role in urination]. PMID- 2573137 TI - Modulation of transmitter and hormone release by multiple neuronal opioid receptors. PMID- 2573139 TI - [Tooth injuries. Diagnosis, treatment]. PMID- 2573138 TI - [Effect of renovascular roentgen contrast media on the function of the healthy kidney--an individual comparison]. AB - In 50 patients with healthy kidneys the excretion of tubular enzymes was determined quantitatively before and after administration of an ionic and a non ionic contrast medium and compared intra-individually. Both preparations produced a highly significant increase in enzyme levels to about twice the initial value, the increase being significantly lower with the non-ionic contrast medium. This effect was fully reversible within 24 hours. No noticeable change was seen within the examination period of 48 hours in respect of creatinine and creatinine clearance levels. PMID- 2573141 TI - [Second international symposium on cardiac insufficiency organized by ICI Pharma (Monte Carlo, May 10 and 11th 1989)]. PMID- 2573140 TI - [Periarteritis nodosa linked to the hepatitis B virus]. AB - Periarteritis nodosa is a necrotizing vasculitis which, in more than one-third of the cases, is consecutive to an hepatitis B virus infection. This aetiological form is usually characterized by the absence of respiratory manifestations and by a greater frequency of hepatic cytolysis. In terms of survival, its prognosis is similar to that of other forms, but a persistent viral replication facilitates the subsequent development of chronic liver diseases and cirrhosis. Its modern treatment consists of a short course of corticosteroids combined with antiviral agent and plasma exchanges. Long-term corticosteroid therapy and immunodepressants are reserved for cases where the above-mentioned treatment has failed. This therapeutic approach results in a survival rate of 80 p. 100 at 4 years. PMID- 2573142 TI - [Food preferences and domiciliation of Culicidae mosquitoes in the Ribeira Valley, Sao Paulo, Brazil, with special reference to Aedes scapularis and Culex (Melanoconion)]. AB - New results on blood-meal identification and the environmental distribution of mosquitoes collected in four different Ribeira Valley (S. Paulo State, Brazil) environments, during the period February to November 1986, are presented. Sources of 651 blood-meals were identified. The preference of Ae. scapularis for large mammals, chiefly cattle, horse and even man, was confirmed. Data suggests that a similar behavioral pattern is presented by Cx. ribeirensis. Both mosquitoes seem to be strongly attracted by peridomiciliar blood sources represented by domestic animals sheltered in that environment. Nevertheless, the female of Ae. scapularis females may use the extradomiciliary environment, represented by residual forests, as resting place in the same way as it uses the peridomiciliary one. The data confirm the clear tendency of both Ae. scapularis and Cx. ribeirensis to adapt to the man-made environment and thus the evolution towards domiciliation in the behavior of these mosquitoes. Other culicids, such as An. bellator, An. cruzii and Cq. chrysonotum, were collected practically only by the use of human bait. For An. cruzii the number of females collected by this method represented 31.6% of the female total collected during this project. Some data about other species are given, and the need for further researches about Culex (Melanoconion) species is emphasized. PMID- 2573143 TI - Gastric and esophageal acidity during continuous treatment with H2-antagonists in uncomplicated esophagitis. AB - Twenty-four-hour gastric and esophageal pH were monitored simultaneously in 19 patients with moderate esophagitis before and after a randomized crossover treatment with 40 mg famotidine or 300 mg ranitidine. Gastroesophageal reflux in patients with esophagitis was compared with that in 22 healthy controls. Patients with esophagitis had more esophageal acidity than controls; the percentage of time with esophageal pH less than 4 was significantly greater during a 24-h period (p less than 0.01) both in the upright (p less than 0.01) and in the supine (p less than 0.01) position. In esophagitis patients the percentage of time with pH less than 4 during the total 24-h period correlated closely with acid reflux in the upright (p less than 0.001) and supine (p less than 0.01) position. This indicates that daytime reflux is probably as injurious to the esophagus as nighttime reflux. Famotidine and ranitidine decreased gastric acidity during the entire 24-h period (p less than 0.001) but not during the daytime or early evening. The inhibitory effect lasted slightly longer with famotidine (12 h) than with ranitidine (10 h). Famotidine and ranitidine reduced esophageal acidity during the entire 24-h period (p less than 0.001) and particularly during the nighttime (p less than 0.001) but not during the daytime. Famotidine and ranitidine also did not modify the esophageal acid clearance. Nightly doses of famotidine or ranitidine were ineffective in reducing GER during daytime hours. PMID- 2573144 TI - Clinical relevance of sampling rate in the characterization and analysis of 24 hour gastric acidity. A report on 413 cases. AB - Four hundred and thirteen continuous, high-frequency-sampled gastric pH-metries were subdivided into six groups (normal subjects and patients with duodenal ulcer, gastric ulcer, chronic gastritis, prior cholecystectomy, or antisecretory treatment). The frequency distribution of those pH fluctuations that were greater than the quantitation error of instruments did not differ significantly (p greater than 0.30) between groups. Five per cent of these real pH fluctuations lasted less than 3 min. As the sampling rate increased from 1 to 60 min, the percentage of subjects whose 24-h median of pHs differed by no more than +/- 10% from that calculated on the raw high-frequency-acquired data progressively decreased (99.8%-42%). This error propagation was more marked in the case of [H+] integrals (99%-10%). A sampling rate equal to or faster than 1 min is necessary to provide a proper representation of the circadian pH pattern and to calculate accurate acidity indexes, regardless of the physiologic or pharmacologic nature of the profile. PMID- 2573145 TI - The potential impact of Campylobacter pylori on the treatment of duodenal ulcer disease. AB - In most studies the association between Campylobacter pylori infection, Type B gastritis and duodenal and gastric ulceration is extremely strong. The best evidence for it having an aetiological role is at present in Type B gastritis. It should be remembered, however, that serological studies show carriage of the organism to be common also in the general population. An attempt is made here to gain some idea of the clinical impact of C. pylori infection in duodenal ulcer disease by analysing clinical trials, and in particular relapse data, in which an agent which suppresses C. pylori is used (colloidal bismuth subcitrate) and compared with one which does not (an H2-receptor antagonist). A mathematical model based on the data from these studies predicts that the prevalence of active duodenal ulceration will be twice as common in a group of subjects treated repeatedly upon relapse with an H2-receptor antagonist as in a group treated with colloidal bismuth. Other possible mechanisms are discussed but the ability of bismuth to suppress C. pylori infection is perhaps the best available explanation at present. Early data from several centres suggest that patients who can be rendered C. pylori negative over a prolonged period of time are relatively immune from relapse of their duodenal ulcers. If confirmed this observation may well transform the long-term management of duodenal ulcer disease. PMID- 2573146 TI - Mucosal protection by H2 antagonists against injury by non-steroidal anti inflammatory agents. AB - It is difficult to ascertain the incidence of gastrointestinal side effects associated with intake of non-steroidal anti-inflammatory drugs (NSAIDs). Serious side effects, such as bleeding, perforation, and heart failure, occur in approximately 1% of patients using NSAIDs. One-third of all patients receiving NSAIDs will have gastrointestinal complaints. Since at least 10% of patients terminate treatment with NSAIDs as a result of side effects, even reduction of those that are not life-threatening would be of great benefit. H2-receptor antagonists have proved effective in ulcer treatment, and their use as prophylaxis against the side effects of NSAIDs is being widely studied. In a recent study, 63 patients who had experienced serious upper gastrointestinal side effects were given cimetidine while continuing their NSAID therapy. All but 4 of 47 who had gastric or duodenal ulcer on first admission were healed at 8 weeks, and none of the remaining 16 with diffuse bleeding gastritis experienced further clinical episodes of bleeding or ulcer-related dyspepsia. Bijlsma has reported a double-blind multicentre study using cimetidine or placebo. When GI symptoms led to endoscopic evaluation, GI symptoms were significantly reduced after cimetidine. PMID- 2573148 TI - Advances in the genetics of schizophrenia: editors' introduction. AB - In view of the rapid advances in the application of molecular genetics to research in schizophrenia, an effort was made to put these studies into a wider perspective. This issue of the Schizophrenia Bulletin is devoted to the expanded presentation of some of the early genetic linkage findings in schizophrenia, and a broader discussion of the impact of these new techniques on schizophrenia research, as well as the limitations of this approach in a complex disorder. PMID- 2573147 TI - Influence of chronic rioprostil treatment on gastric endocrine function. AB - The influence of 4 weeks of treatment with the prostaglandin E1 analogue, rioprostil, 300 micrograms b.i.d., or placebo, on gastric endocrine function is tested in healthy male volunteers. Basal serum gastrin levels and postprandial gastrin output are unchanged after treatment with rioprostil. Similarly, plasma pancreatic polypeptide levels are unaffected. Antral gastrin tissue concentrations as well as antral somatostatin concentration and volume densities of antral G-cells and D-cells are unchanged. Neither basal nor pentagastrin stimulated gastric acid secretion after rioprostil therapy differ from pretreatment values. Rioprostil, given in a dose of 100 micrograms b.i.d. to rats for 1 week significantly increases antral mucosal height but has no influence on the mucosal concentrations and cell densities of the gastric peptides, gastrin and somatostatin. It is concluded that rioprostil in the dose used does not affect the endocrine stomach after 4-weeks' administration at a dose of 300 micrograms b.i.d. PMID- 2573149 TI - Molecular genetic studies in schizophrenia. AB - Despite many years of research, the genetic factors in schizophrenia are not well understood. Recent developments in DNA technology allow new methods of testing genetic hypotheses in the etiology of this debilitating disorder. We have found evidence against linkage of schizophrenia in a Swedish kindred to markers on chromosome 5; another research group has reported positive evidence for linkage to this same chromosomal region in British and Icelandic families. This article presents a set of data expanded from our previous report, discusses the issue of heterogeneity, and reviews the current status of linkage studies in schizophrenia. PMID- 2573151 TI - Old gene, new trick. PMID- 2573150 TI - Diurnal weight gain in chronic psychosis. AB - We found diurnal weight gain to be abnormal among 93 chronically psychotic patients, most of whom had schizophrenia. They were weighed at 7 a.m. and 4 p.m. weekly for 3 weeks. We normalized the diurnal weight gain (NDWG) as a percentage by subtracting the 7 a.m. weight from the 4 p.m. weight, multiplying the difference by 100, and dividing the result by the 7 a.m. weight. NDWG was 1.7 +/- 1.0 percent for the study sample, 0.6 +/- 0.4 percent for 16 acutely psychotic controls, and 0.5 +/- 0.4 percent for 29 normals. More than 60 percent of the study sample had abnormal NDWG values. NDWG related to antipsychotic drug dose (r = 0.290, p = 0.005) with variability in drug dose accounting for 8 percent of the variability in NDWG. This report provides yet another piece of evidence that disordered water balance is common in chronic psychiatric patients. The etiology is unknown, but it may relate to subtle brain abnormalities in the regulation of fluid intake and excretion. PMID- 2573152 TI - A biochemical correlate of the critical period for synaptic modification in the visual cortex. AB - Stimulation of phosphoinositide hydrolysis by excitatory amino acids was studied in synaptoneurosomes of kitten striate cortex at several postnatal ages. Ibotenate and glutamate stimulated phosphoinositide turnover during the second and third postnatal months; N-methyl-D-aspartate and DL-alpha-amino-3-hydroxy-5 methyl-4-isoxazole propionic acid (AMPA) were without effect. The developmental profile of ibotenate-stimulated phosphoinositide turnover parallels the postnatal changes in cortical susceptibility to visual deprivation. The transient increase in ibotenate-stimulated phosphoinositide turnover does not occur in visual cortex of kittens reared in complete darkness. PMID- 2573153 TI - Tonic activation of NMDA receptors by ambient glutamate enhances excitability of neurons. AB - Voltage clamp recordings and noise analysis from pyramidal cells in hippocampal slices indicate that N-methyl-D-aspartate (NMDA) receptors are tonically active. On the basis of the known concentration of glutamate in the extracellular fluid, this tonic action is likely caused by the ambient glutamate level. NMDA receptors are voltage-sensitive, thus background activation of these receptors imparts a regenerative electrical property to pyramidal cells, which facilitates the coupling between dendritic excitatory synaptic input and somatic action potential discharge in these neurons. PMID- 2573154 TI - Effect of carboxylic acid side chains on the absorption maximum of visual pigments. AB - The proposal that the absorption maximum of the visual pigments is governed by interaction of the 11-cis-retinal chromophore with charged carboxylic acid side chains in the membrane-embedded regions of the proteins has been tested by mutating five Asp and Glu residues thought to be buried in rhodopsin. Changing Glu113 to Gln causes a dramatic shift in the absorption maximum from 500 nanometers to 380 nanometers, a decrease in the pKa (acidity constant) of the protonated Schiff base of the chromophore to about 6, and a greatly increased reactivity with hydroxylamine. Thus Glu113 appears to be the counterion to the protonated Schiff base. Wavelength modulation in visual pigments apparently is not governed by electrostatic interaction with carboxylate residues, other than the counterion. PMID- 2573155 TI - [The possibilities of osteoplastic measures in mid-foot amputation and osteomyelitis of the tarsus]. AB - Osteomyelitis after trauma or after operations and amputations at the midfoot or tarsus leads to a disadvantage in terms of motion and of weightbearing on the foot. Once local fistula, exophytic bone growth, skin defects and instability of the metatarsus have arisen, hygiene and recovery of joint function are difficult. Four patients were treated by reamputation at the midfoot and bone autografts. The osteomyelitic part was excised and the tarsal and metatarsal bones stabilized. The result at follow up showed painfree stumps without recurrence of infection within 3 years. Little support is needed in the way of orthopedic appliances, and all four patients can walk easily with no pain. This operation is recommended for the primary surgical procedure. PMID- 2573156 TI - Case report 568: Total resorption of the lateral sesamoid secondary to Pseudomonas aeruginosa osteomyelitis. PMID- 2573157 TI - [Complications of the treatment of rheumatoid arthritis with chrysanol]. PMID- 2573158 TI - [Hormonal disorders in borderline arterial hypertension]. AB - Measurements of blood plasma ACTH, hydrocortisone, STH, somatostatin, insulin, glucagon levels and plasma renin activity in 70 patients with borderline hypertension (BAH) and in 20 normal male subjects have revealed increased ACTH, hydrocortisone, and somatostatin levels, elevated plasma renin activity, and reduced STH and insulin levels in the patients. A possible role of the pressor hormone system activation in the pathogenesis of borderline arterial hypertension and in BAH transformation into essential hypertension is discussed. PMID- 2573159 TI - [Effect of salazosulfapyridine and its metabolites on immunocompetent cells]. AB - Long-term administration of SASP does offer clinical benefit and has a demonstrable disease modifying effect in rheumatoid arthritis, though its mode of action remains obscure. We have studied the in vitro effects of SASP and its metabolites, that is SP, ASA, AcSP and AcASA, on the blast-formation of lymphocytes, the cytotoxic activity of NK cell, the phagocytosis and H2O2 production of monocyte and the fMLP-induced chemotaxis and superoxide anions production of PMNs. We have obtained the following results: (1) the blast formation of lymphocytes by PHA and Protein A was significantly inhibited by SASP, but not by the metabolites; (2) the cytotoxic activity of the NK cell was inhibited by SP and AcSP, but not by SASP, ASA and AcASA; (3) on monocyte, SP, AcSP and AcASA inhibited phagocytosis, and all of drugs had no effect on the production of H2O2; (4) on PMNs, SASP, SP and ASA significantly inhibited fMLP induced chemotaxis, and SASP and all of its metabolites significantly inhibited a release of superoxide anions by stimulation of fMLP and PMA; (5) SASP and ASA scavenged superoxide radical at the concentration comparable to clinical doses. In vivo, the above effects may be exhibited in proportion to each blood concentrations of drugs. In particular, it appears that SASP, SP and AcSP play an important role in the therapeutic efficacy in rheumatoid arthritis. PMID- 2573160 TI - Early total thyroidectomy in patients with multiple endocrine neoplasia IIb syndrome. AB - Medullary carcinoma of the thyroid gland (MCT) develops at an early age in children with multiple endocrine neoplasia (MEN) IIb syndrome. Prompt diagnosis of the syndrome and early total thyroidectomy offer the only chance for cure. Surgical guidelines for managing the pediatric patient with potential for MCT have included screening with provocative calcitonin tests beginning at one year of age and total thyroidectomy performed when either basal calcitonin or stimulated test levels become abnormal. The diagnosis of MEN IIb, however, can be made on the basis of pathognomic clinical features, including characteristic facies, marfanoid body habitus, submucosal ganglioneuromas and thickened corneal nerves. A family is described herein in which one member had the pathognomonic clinical findings of MEN IIb but did not have an elevated level of serum calcitonin in response to stimulation with pentagastrin and calcium. Nonetheless, a total thyroidectomy was performed and multiple microscopic foci of MCT were found on pathologic examination. This suggests that total thyroidectomy should be performed upon any patient with the characteristic phenotype of MEN IIb, regardless of the results of the stimulation test for calcitonin. PMID- 2573162 TI - [National Congress. Welcome to the West!]. PMID- 2573161 TI - [Design and effect of sports shoe insoles for the runner]. AB - Running injuries of the lower extremities are more frequent with increasing intensity and extent of training. Biomechanical investigations demonstrate that exertion induced injuries of the foot, achillotendinitis, patellar tendinitis and shin splint syndrome are correlated with disorders in footstrike patterns. Therefore running shoe insoles can eliminate peak forces by exactly placing supporting elements. Furthermore peak forces may be distributed and complaints may be reduced. 50 runners with exertion induced injuries of the lower extremity were provided with appropriate running shoe insoles. There were excellent results in 34% and good results in 42%. 20% had moderate improvements while only 4% had unchanged complaints. PMID- 2573163 TI - [Variation in the urinary excretion of 6-beta-hydroxycortisol in humans after administration of the new isoquinoline derivative, PK-11195 (52028 RP)]. AB - Among in vivo tests to assess liver drug metabolizing enzyme induction, urinary 6 beta-hydroxycortisol (6-beta-OHF), plasma gamma-glutamyltransferase (GGT) and urinary D-glucaric acid are most frequently proposed. 6-beta-OHF is the most abundant unconjugated metabolite of cortisol in human urine. We measured its elimination during a clinical trial in 16 human healthy volunteers (men and women), these persons being treated by a new isoquinoleine derivative, 52028 RP (PK-11195). This drug is an antagonist of peripheral type benzodiazepine binding sites. Urinary excretion of 6-beta-OHF increased significantly (3.5 fold, p less than 0.01) on the 5th day of treatment (400 mg/day, orally) and remained increased as long as the treatment was continued (15 days). Control values were again observed 5 days after stopping the treatment. Plasma gamma glutamyltransferase activity and D-glucaric acid urinary elimination are increased more than 2 fold. The data demonstrated that 6-beta-OHF in the most sensitive among the three tests performed to detect a drug metabolism induction, during this clinical trial. PMID- 2573165 TI - Coated pits and vesicles transfer plasma components to platelet granules. AB - Electron microscopy of mammalian blood platelets after contrast enhancement with tannic acid after an initial fixation in glutaraldehyde and osmium reveals numerous coated pits (c.p.) and vesicles (c.v.), indicating a process of receptor mediated endocytosis. The c.p. may be located at any site of the plasma membrane or the canalicular, surface connected membrane system. C.v. fuse with platelet granules without losing their coat. Evidence for a continuous transfer of ambient fluid to granules via c.p. and c.v. was obtained by the use of fluid-phase markers. It is proposed that the endocytic process may play a role in blood platelet activation. PMID- 2573166 TI - [G proteins: GTP binding proteins in signal transduction]. AB - A family of GTP-binding proteins (G proteins) couples receptors for hormones and neurotransmitters to diverse effectors in the cell. G proteins are membrane-bound GTPases composed of three distinct polypeptides. Cyclic AMP production, phosphoinositide breakdown, and ion channels are all known to be regulated via G proteins. Functional characterization, protein purification, labelling with bacterial toxins, specific antibodies and molecular cloning are tools which have helped to provide information on these proteins. Although the molecular properties of several G proteins are well known, the correct functional assignments are complex. Furthermore, we do not know how the various signal systems are involved in regulation of G protein activity. PMID- 2573164 TI - Synthesis of vitamin K-dependent proteins by cultured human tumor cells. AB - The observation that warfarin inhibits the growth and metastasis of certain types of clinical and experimental tumors suggests a role for vitamin K in tumor biology. We have investigated synthesis of vitamin K-dependent proteins in four malignant (lung epidermoid carcinoma, melanoma, colon adenocarcinoma, and breast adenocarcinoma) and three normal (colon epithelium, breast epithelium, and fibroblasts) cell lines of human origin grown in tissue cultures. Our results show the following: 1) Vitamin K-dependent carboxylase activity is present in all of the malignant and normal cell lines studied. 2) The malignant as well as normal cell lines synthesize a family of vitamin K-dependent proteins. Microsomal precursors of these proteins with apparent molecular mass of 74, 62, and 34 kDa are common to all malignant and normal cell lines whereas precursors of higher and lower molecular mass seem to be synthesized by some but not all tumor cell lines. 3) The 74 kDa precursor synthesized by colon carcinoma and breast carcinoma was positively identified as a precursor of protein S. PMID- 2573167 TI - [Beta blockaders and physical performance. Limiting factors]. AB - Beta blockers are known to cause reduced exercise performance in hypertensive and healthy subjects. Additive effects of selective blockade of beta-1 and beta-2 receptor subtypes seems to account for the total reduction in exercise capacity observed with non-selective beta-1,2 blockade. The mechanism is as yet undefined. The magnitude of the reduction is dependent of type of exercise and level of fitness. Hemodynamic parameters, substrate delivery to the working muscles, mental factors, and interference with potassium homeostasis may be involved. PMID- 2573168 TI - [New aspects of the pathogenesis of ischemic brain damage. Possible involvement of excitatory amino acids]. AB - Glutamate, a major excitatory transmitter substance, is neurotoxic at high concentrations. Brain dialysis experiments have demonstrated an extracellular overflow of glutamate during ischemia, and there is good evidence from several animal models that glutamate antagonists offer partial protection against the development of ischemic cell degeneration. These and other experimental data indicate that glutamate may be involved in the pathogenesis of ischemic brain damage. Quantitative immunocytochemical investigations carried out in the authors' laboratory suggest that ischemia is associated with loss of glutamate from nerve cell bodies, and reduced ability of the glial cells to metabolize glutamate. We discuss possibilities of new therapeutic strategies. PMID- 2573169 TI - NcoI restriction fragment length polymorphism (RFLP) of the tumor necrosis factor (TNF alpha) region in four autoimmune diseases. AB - The two-allele NcoI Restriction Fragment Length Polymorphism (RFLP) of the TNF alpha region yielding bands of 5.5 and 10.5 kb was investigated in patients with systemic lupus erythematosus (SLE), pauciarticular juvenile rheumatoid arthritis (P-JRA), rheumatoid arthritis (RA) and primary Sjogren's syndrome (pSS). In all four disease, we found a decreased frequency of the 10.5 kb allele which, however, was significant only in the SLE group. In all conditions except RA, the frequency of the 5.5 kb fragment was increased. In pSS and SLE, the frequency of HLA-B8 was increased in 5.5 kb fragment positive patients compared with corresponding controls and thus, the increase of this band and the decrease of the 10.5 kb band may be secondary to HLA-B8 associations owing to strong positive linkage disequilibrium between HLA-B8 and the 5.5 kb band. PMID- 2573170 TI - Individuals with HLA-DR blank alleles display well-known DR-DQ RFLPs. AB - We have characterized HLA-DRB, -DQA, and -DQB gene polymorphism in a large number of serologically DR blank haplotypes with the restriction enzymes Taq I, Bam HI, and Pvu II, with the aim of finding new RFLPs in the Caucasian population. Locus specific RFLPs were combined for a definition of Taq I DR-DQ haplotypes. All observed DNA haplotypes could be found in a control group of 100 individuals, but with a different distribution. Serologically less well-defined specificities were over-represented in the blank group, in particular the DRw13-associated Taq I DR DQ haplotype T-13.3. We conclude that the majority of DR blank haplotypes are probably closely related or identical to previously defined DR alleles. The extent of DR polymorphism in the Caucasian population seems to be well mapped, considering the extremely small proportion of rare Taq I DR-DQ haplotypes and lack of new patterns in this study. PMID- 2573171 TI - [Two cases of colitis cystica profunda in dogs]. AB - Colitis cystica profunda in dogs has been diagnosed in one case only. The two own cases were characterized by repeated, partly bloody diarrhea, vomitus, and painful defecation. The disease was diagnosed by clinical examination and colonoscopy with the ensuing histological examination of biopsy specimens. The disease could be managed by administration of a diet, sulfasalazine and corticosteroids. PMID- 2573172 TI - In vitro accumulation of thyroid hormones by cultured rat hepatocytes and the biliary excretion of iodothyronines in rats treated with a novel histamine H2 receptor antagonist. AB - The administration of SK&F 93479, a novel histamine H2 antagonist, to Wistar rats has been shown to produce thyroid lesions associated with an increased clearance of plasma thyroxine (T4) and elevated plasma TSH concentrations. To determine if these changes were mediated via an increase in the hepatic clearance of thyroid hormones, the biliary clearance and hepatic accumulation of iodothyronines were measured in rats and cultured hepatocytes treated with SK&F 93479. Both the in vitro and in vivo results showed that treatment with SK&F 93479 caused an increased hepatic accumulation (approx. 200% of control) and biliary excretion (2 3-fold control values) of T4 but had little or no effect on T3 uptake. The in vitro studies showed that the treatment related increase in hepatic thyroxine accumulation was temperature and therefore probably energy dependent, while inhibition of cytochrome P-450 dependent enzyme activity did not alter the accumulation of T4 suggesting that the parent compound and not some oxidative metabolite was responsible for the hepatic effects. Chromatographic analysis of bile from SK&F 93479 and phenobarbitone-treated rats showed that in the latter animals 81% of the T4 was present as the glucuronide conjugate (consistent with enzyme induction) whereas in the SK&F 93479-treated rats only 25% was present as the conjugate with 75% being in the unconjugated form. These studies show that SK&F 93479 increases the hepatic accumulation and biliary clearance of T4 by a novel mechanism not associated with liver microsomal enzyme induction. PMID- 2573174 TI - Release of 2-thiobarbituric acid reactive products from glutamate or deoxyribonucleic acid by 1,2,4-benzenetriol or hydroquinone in the presence of copper ions. AB - Cytotoxic effects of various quinone compounds are thought to be due to the formation of semiquinone free radicals. Hydroquinone and 1,2,4-benzenetriol in the presence of copper ions release from glutamate or DNA aldehydic products capable of reacting with 2-thiobarbituric acid (TBA). The formation of TBA reactive products (TBAR) was greater in the presence of 1,2,4-benzenetriol in comparison with hydroquinone. Complete inhibition of formation of TBAR from glutamate by 1,2,4-benzenetriol and copper was observed in the presence of catalase, thiourea and mannitol. Albumin and superoxide dismutase offered substantial protection. Complete protection of formation of TBAR from DNA was observed in the presence of catalase and thiourea. Presence of albumin, mannitol and superoxide dismutase caused only partial inhibition. The formation of TBAR from glutamate or DNA is dependent on copper ion concentration. The present data indicate that hydroquinone and 1,2,4-benzenetriol in the presence of copper ions can lead to the formation of reactive hydroxyl radicals which can release TBAR from glutamate or DNA. PMID- 2573173 TI - Effects of acute and sub-chronic administration of iron nitrilotriacetate in the rat. AB - Parenteral administration of iron nitrilotriacetate (FeNTA) to rats resulted in marked loss in body weight, and increases in liver/and kidney/body weight ratios. Fatalities, due to renal failure, depended on dosage and age of the animals, and were greater (70%) after a single large dose (12 mg iron) than after repeated smaller doses (30%). FeNTA administered subchronically gave rise to an increase in ethane exhalation, and to decreased liver glutathione peroxidase activity, and decreased cytochrome P-450 concentration and benzphetamine N-demethylase activity. It also resulted in severe renal tubular necrosis, with deposition of iron in the tubular cells and loss of brush border alkaline phosphatase activity, resulting in a dose-dependent diuresis, with increased urinary excretion of glucose, iron and lipid peroxidation products, and decreased urine creatinine concentration. NTA alone had none of these effects but slightly decreased the hepatic concentration of iron. PMID- 2573175 TI - Low level lead neurotoxicity in a pregnant guinea pigs model: neuroglial enzyme activities and brain trace metal concentrations. AB - Specific activities of the astroglial marker glutamine synthetase (GS), and the oligodendroglial marker glycerol-3-phosphate dehydrogenase (GPDH) were measured in the spinal cord of fetal guinea pigs and their dams following chronic exposure to low levels of lead (Pb) during gestation. In addition, the effects of Pb on intracellular trace metals (Cu, Fe, Zn) were measured in the blood, cerebellum and forebrain. Aminolevulinic acid dehydratase (ALAD) and zinc protoporphyrin IX (ZPP) were measured in order to monitor established parameters of Pb-exposure. Pregnant guinea pigs were orally administered 0, 5.5 or 11 mg Pb/kg body weight for 30 or 40 days commencing on day 22 of gestation. Blood Pb levels produced in dams and fetuses were at or near the currently identified "no effect" levels for children (10-30 micrograms/dl). These Pb blood levels produced a significant (P less than 0.05) dose-dependent decrease in GS and GPDH activity in the dams and fetuses. Fe and Zn concentrations in blood, cerebellum and forebrain of both dams and fetuses were significantly (P less than 0.05) decreased in a dose-dependent manner. However, Cu concentrations in the blood, cerebellum and forebrain were decreased in the dams but increased in the fetuses in a dose-dependent fashion. The alteration of trace metal concentrations is a proposed mechanism of Pb neurotoxicity. Blood ALAD activity was significantly (P less than 0.05) decreased and ZPP levels were significantly (P less than 0.05) increased, as expected in Pb exposed animals. This study presents the first biochemical evidence for the alteration of neuroglial function at low levels of Pb exposure and focuses attention on the fetus as an important Pb target. PMID- 2573176 TI - Renal and hepatic toxicity of a benzopyran-4-one in the Cynomolgus monkey. AB - The administration of PD 119819, a novel benzopyran-4-one brain dopamine autoreceptor agonist, to Cynomolgus monkeys was followed by deposition of needle like drug crystals in the bile canaliculi, hepatocytes, proximal renal tubules and renal parenchyma. The crystals were associated with a granulomatous inflammation, and histological and biochemical evidence of hepatic and renal cell damage. Although metabolism differences may be the reason why primates, but not rodents, developed these changes, this form of crystallization appeared to be primarily a result of the insolubility of PD 119189 at alkaline pH. PMID- 2573177 TI - Modification of arginine in sea anemone toxin RTX-III from Radianthus macrodactylus. AB - Chemical modifications of the polypeptide neurotoxin RTX-III have allowed us to study the functional role of Arg residues. The effect of chemical modification has been estimated by measuring toxicity in mice. 2,4-Pentanedione did not react with Arg residues of RTX-III even after 100 hr incubation. Malonic aldehyde reacted readily with RTX-III, yielding an unusual derivative; a Schiff's base obtained by condensation of one of two aldehyde groups of malonic aldehyde with the guanidine group. The derivative was one-fourth as toxic as the native toxin. Modification of the guanidine side chain of Arg-13 with both 1,2-cyclohexanedione and phenylglyoxal decreased the toxicity of RTX-III by a factor of five. We conclude that Arg-13 is not fully responsible for toxicity. The toxin-receptor attachment might be multipoint, involving several structural elements of the protein molecule, with Arg-13, probably being one of them. The guanidine side chain of Arg-45 is buried in the sequence and apparently functionally nonessential. PMID- 2573178 TI - Steady-state fluctuation and variability of betaxolol and atenolol plasma levels. AB - Twenty-four nonsmoking male volunteers took 50 mg atenolol or 10 mg betaxolol orally once a day for 9 days in a two-period, four-sequence, randomized, crossover study. Plasma concentrations reached steady state after day 5. Percent fluctuation in plasma concentration defined as (Cmax-Cmin)/Cavg (% fluctuation 1) was 97% on day 9 for betaxolol and 343% for atenolol; thus atenolol fluctuation was more than threefold that of betaxolol. A 10-fold difference in plasma level fluctuation was observed when fluctuation was defined as (Cmax-Cmin)/Cmin (% fluctuation 2). The intersubject variances for % fluctuation 1 and % fluctuation 2 were 4.1 and 85.5 times greater for atenolol than for betaxolol; these differences were marginally statistically significant for % fluctuation 1 and significant for % fluctuation 2. The intrasubject variabilities for area under the curve and plasma level fluctuations were statistically greater for atenolol than for betaxolol. Atenolol intrasubject variances were 25 and 271 times greater than for betaxolol for % fluctuation 1 and % fluctuation 2, respectively. Thus, betaxolol exhibited less fluctuation in plasma levels with substantially less intersubject and intrasubject variability. These factors would be expected to provide a more consistent therapeutic response and more dependable dosage adjustment. PMID- 2573179 TI - Function and surface phenotype of T lymphocytes infiltrating renal allografts in nonhuman primates treated with monoclonal antibodies. AB - The phenotype and function of T lymphocyte cell lines established in vitro from kidney biopsies at the time of acute cellular rejection were studied using a nonhuman primate renal allograft model. Our objectives were to investigate the function and surface phenotype of cells that infiltrate renal allografts in animals that were untreated, that were given subtherapeutic cyclosporin, or that developed rejection after treatment with monoclonal antibodies to IL-2R B chain (CD25), immune cell adhesion molecule-1 (ICAM-1), or CD8. Lines from allograft biopsies and peripheral blood were expanded in vitro using solely human recombinant IL-2 and analyzed after 6-20 days in culture. We found that the large majority of cells cultured from cynomolgus allografts at the time of acute rejection or, when possible, assayed directly without culture, were CD3+4-8+ T lymphoblasts that possessed donor-specific cytolytic function and an NK-line, cytotoxic activity. In contrast, it was rarely possible to establish T cell lines exhibiting donor-specific cytotoxic activity from the blood except in the absence of immunosuppression or during CsA taper. A stable number of graft-derived CD4+8- cells was only observed in an unsuppressed animal 2 days after transplantation in the absence of manifest signs of rejection. Taken together, the above data indicate that similar T lymphocyte populations associated with allograft rejection are present in acutely rejecting allografts after the various types of immunosuppressive therapy. Since the infiltrating cells were similar to those obtained prior to therapy, recurrent rejection most likely represents cells that have escaped elimination. The T cells derived from monkey grafts differ from those from human renal allografts by the decreased frequency of CD4+ cells. Whether this difference is species-related or therapy-related is not known. PMID- 2573180 TI - Cardiac allograft survival across major histocompatibility complex barriers in the rhesus monkey following T lymphocyte-depleted autologous marrow transplantation. III. Late allograft rejection. AB - We have studied organ allograft survival in rhesus monkeys conditioned with myeloablative total-body irradiation and T cell-depleted autologous bone marrow transplantation then given a heterotopic MHC-mismatched cardiac allograft in the immediate postmyeloablative period. This model has enabled us to investigate the role of T cells in vascularized organ allograft rejection. We previously reported (1) that recipients of marrow depleted of T cells below a critical threshold (0.16% residual marrow T cells, or 0.14 x 10(5) infused T cells/kg) experienced a period of freedom from acute rejection associated with a profound nonspecific immune deficiency (determined by skin grafting). Resolution of the nonspecific immune deficiency was associated with late graft rejection. In the present report, we correlate the results of peripheral immune reconstitution studies and direct immunohistochemical analysis with allograft status in order to study T cell subsets involved in late rejection. We report that, in contrast with CD8+/CD28- T cells, CD16+ NK cells, and CD20+ B cells, late allograft rejection was associated with the return of peripheral CD4+ T cells and CD8+/CD28+ T cells, suggesting a critical role for one or both of these subsets in late allograft rejection in this model. PMID- 2573181 TI - Cardiac allograft survival across major histocompatibility complex barriers in the rhesus monkey following T lymphocyte-depleted autologous marrow transplantation. IV. Immune reconstitution. AB - Studies of postmyeloablative immune reconstitution have been reported for allogeneic bone marrow transplantation and also for non-T cell-depleted autologous/syngeneic BMT. However, there is a paucity of information regarding immune recovery following T cell-depleted autologous/syngeneic BMT. We have developed a primate transplantation tolerance model in which rhesus monkeys were conditioned with total-body irradiation and extensively T cell-depleted autologous BMT and given a major histocompatibility complex-mismatched heterotopic cardiac allograft. This model provided an opportunity to study peripheral immune recovery following T cell-depleted autologous BMT. Limiting dilution analysis was used to quantify marrow T cells following depletion (2.8% to 25.6% marrow T cells predepletion, 0.00014% to 0.036% residual marrow T cells postdepletion). We found that (1) hematopoietic engraftment was prompt despite extensive marrow T cell depletion, (2) reconstitution of CD4+ helper T cells and CD8+ cytotoxic T cells were substantially delayed (6-12 months) compared with the recovery of CD8+ suppressor T cells, CD16+ NK cells, and CD20+ B cells, (3) distinction between CD8+ cytotoxic T cells and CD8+ suppressor T cells by the CD28 marker was critical in revealing the markedly discrepant recoveries of those subsets, and (4) immune reconstitution resembled that observed in recipients of T cell-depleted allogeneic and non-T cell-depleted autologous/syngeneic BMT, suggesting that the pattern of immune recovery following BMT is not substantially influenced by either allogeneic effects or the number of transferred T cells over a range of values. PMID- 2573182 TI - Pregnancy as a natural model of allograft tolerance. Interactions between adherent macrophages and trophoblast populations. AB - From an immunologic viewpoint, the fetus with its paternal antigens may be considered a successful allograft in the maternal host. Understanding the basis of this host-allograft relationship remains a fundamental unsolved problem in transplantation immunobiology. We have previously demonstrated that local immunoregulation in the murine placenta prevented macrophage activities necessary for an effective response against the intracellular bacterium Listeria monocytogenes. Given the central role of macrophages both as antigen-presenting and cytolytic effector cells, such local immuno-regulation may ordinarily help prevent rejection of the fetoplacental unit with its paternal alloantigens by the maternal immune system. We therefore examined two types of interaction between macrophages and the placental cells that populate the maternal-fetal interface. (1) Upon activation to kill listeria efficiently, macrophages also acquire cytolytic capacities against some tumor and embryonic cells. We tested the hypothesis that macrophage activation in the placenta was inhibited to prevent macrophages from lysing fetal trophoblasts. We found, however, that trophoblasts isolated by dispase dispersion, differential isopyknic centrifugation, and adherence were not lysed by three different populations of cytolytic macrophages: (a) those activated in vivo during listeriosis, (b) peptone-elicited macrophages activated in vitro by recombinant interferon gamma and other lymphokines, and (c) the macrophage cell line RAW 264.7 activated in vitro. (2) Previous studies had demonstrated that cells from the placental region and their conditioned media inhibited a variety of lymphocyte functions. However, we found that these did not inhibit activation of adherent macrophages as assessed by induction of cell surface Ia and acquisition of tumoricidal activity. In addition, under conditions where placental cells inhibited the proliferative response of a cloned CD4+ anti Listeria T cell line to fixed, antigen-pulsed macrophages, the secretion of macrophage-activating lymphokines was not affected. These studies are important because they indicate that previously described suppressor systems in the murine placental region do not account for the profound local deficits in macrophage function seen during listeriosis. PMID- 2573183 TI - Restriction fragment length polymorphism analysis of canine class II major histocompatibility complex genes. PMID- 2573185 TI - Reversal of diabetes in CD4+ T cell-depleted mice by xenotransplantation of pig proislets. PMID- 2573184 TI - Anti-T cell-specific immunoconjugates in mice: in vivo effects. PMID- 2573186 TI - Restriction fragment length polymorphism analysis of class II genes and canine bone marrow transplantation: correlation with mixed lymphocyte reactivity. PMID- 2573187 TI - CD13: a signal transduction molecule on myeloid cells? PMID- 2573189 TI - Onco-suppressor genes in human cancer. AB - The analysis of the molecular mechanisms governing multistep carcinogenesis became experimentally approachable since the identification and characterization in tumor cells of altered or activated versions of cellular genes (oncogenes) that normally control cell growth and differentiation. The activating mutations confer new properties to the oncogene products and should therefore be considered as gain of function mutations. In addition, the oncogenes appear to act as dominant genetic traits since they act also in the presence of the homologous wild-type allele. However, the concept of a dominance of the transformed phenotype has been challenged by early experiments with somatic cell hybrids which showed that the fusion of normal and malignant cells may suppress the tumorigenic phenotype. The suppression or reversion of the malignant phenotype by the introduction of a normal chromosome into a tumor cell line has lent support to the idea that a family of cellular genes are coding for factors capable to interact with the cell-growth control machinery. These genes seem to reconstitute the normal control of cell growth even in the presence of an activated oncogene. In addition, a two-mutation model has been proposed to explain the epidemiological and clinical features of childhood cancers. According to the model, the development of these malignancies can be caused by the loss or inactivation of both alleles of cellular genes, as suggested by the somatic cell hybrid experiments where the function of the inactivated genes is restored by the contribution of those derived from the normal parental cells. This family of genes is designated as onco-suppressor genes since their product is necessary for the normal regulated cell growth and is lacking or inactivated in malignant cells. At gene level they should be considered as recessive genetic traits, since the tumor phenotype appears when both alleles of an onco-suppressor gene are inactivated. The mutations affecting their normal functions belong to the type "loss of function". The molecular analysis of retinoblastoma has led to the cloning and sequencing of the related onco-suppressor gene (RB gene) whose product displays the features of a gene-regulatory protein. In addition, a binding between the RB product and various viral onco-proteins (E1A, large T, E7) has been demonstrated, thus suggesting a mechanism of RB inactivation by which some DNA viruses can transform the host cell.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2573188 TI - Beta blockers in portal hypertension--current status. AB - Pharmacotherapy of portal hypertension started with the use of intravenous vasopressin in the 1960s. However, the short half life of the drug and the non availability of an oral preparation limited its use to acute variceal bleeding. It was two decades later that propranolol, a beta blocker was shown to decrease portal pressure. However, the usefulness of beta blockers in the treatment of recurrent variceal bleeding remains controversial with controlled trials showing both benefit and lack of benefit. This review is aimed to bring into perspective, current knowledge of the effect of beta blockers on the splanchnic circulation in man and animals, its clinical usefulness in portal hypertension and its place in the therapeutic armamentarium against portal hypertension. PMID- 2573190 TI - Loss of heterozygosity of DQ alpha gene in human malignant melanoma. AB - DNAs from human melanoma cells, used at early in vitro passage, were evaluated by Southern blot analysis for somatic loss of heterozygosity at the DQ alpha gene. A total of 7 melanomas, 3 primary and 4 metastatic derived from 5 different patients were studied; in one case (pt 665) cell lines were derived from two anatomically different subcutaneous metastasis, whereas in a second case (pt 9923) both the primary tumor and a lymph node metastasis were available. Restriction length polymorphism (RFLP) analysis, performed on autologous peripheral blood lymphocyte (PBL) DNA digested with different enzymes, showed a pattern of bands compatible with the constitutional heterozygous typing at DQ alpha gene in 4 cases whereas 1 case revealed an homozygous typing. When melanoma DNAs were analysed, 1 out of the 4 informative cases (pt 1007) showed a loss of a diagnostic fragment for DQ alpha gene when digested with both Taq I and Bgl I enzymes. These results indicate that class II allelic losses detectable by RFLP can be found on malignant melanoma and add further complexity on the involvement of chromosome 6 whose cytogenetic abnormality are the most consistent in this human neoplasia. PMID- 2573191 TI - [Early detection of a contralateral second carcinoma in patients with testicular tumors caused by testicular carcinoma in situ]. AB - Carcinoma in situ (CIS) of the testis is considered to be a precursor of germ cell cancer. Diagnosis is made by the conventional biopsy technique. Only for patients at risk is a screening biopsy justifiable. This group includes patients with testicular cancer in whom the incidence of contralateral second germ cell tumors is increased. In a prospective study we found three cases of testicular CIS in biopsies from the contralateral testes of 61 such patients. All cases with a diagnosis of CIS presented with testicular atrophy (volume less than 12 ml), associated with necrozoospermia in one patient and with azoospermia in two patients. Treatment consisted in local irradiation (20 Gy) of the remaining testis to preserve Leydig cell function. In control biopsies no evidence of CIS or germ cells was found. More than 3 months after therapy, plasma testosterone levels were normal and LH and FSH levels were increased. None of the patients with negative biopsy (n = 49) who were followed up was found to have a second cancer of the contralateral testis. The average observation time so far is 17.2 months. PMID- 2573193 TI - What is new and where are the problems in research on hemorrhagic fever with renal syndrome? PMID- 2573192 TI - Immunohistochemical studies on oncogene products (c-erbB-2, EGFR, c-myc) and estrogen receptor in benign and malignant breast lesions. With special reference to their prognostic significance in carcinoma. AB - It is a matter of debate whether the amplification of c-erbB-2 oncogene or production of the oncoprotein in breast cancers correlate with the presence of lymph node metastasis and with a poor prognosis. This study was aimed at elucidating the immunohistochemical localization of oncogene products which are related to cell growth, c-erbB-2 product, epidermal growth factor receptor (EGFR), c-myc protein and estrogen receptor (ER), in benign and malignant lesions of the breast. Fresh frozen sections of 25 breast cancers and 11 fibroadenomas from Japanese women were studied by indirect immunoperoxidase method with proper fixation. C-erbB-2 product and EGFR were localized on the cell membrane whereas c myc protein and ER were observed in the nuclei. Immunohistochemical expression of oncogene products and ER were not only observed in the mammary carcinomas but also in the fibroadenomas. However immunoreactivities of EGFR and ER were more frequently seen in the fibroadenomas (p less than 0.05). In breast cancers, the incidence of immunoreactivity for c-erbB-2 was higher in the cases with lymph node metastasis than cases without nodal metastasis (p less than 0.05) and there was reciprocal correlation between the expressions of EGFR and ER (p less than 0.05). Regarding the size of the primary tumour, there was no statistically significant correlation with the expressions of c-erbB-2, EGFR, c-myc or ER. Histological grade correlated only with the expression of ER (p less than 0.05). PMID- 2573194 TI - [Bopindolol does not affect normal glucose and lipid metabolism in hypertensive patients]. AB - The authors administered to ten patients with mild to medium severe hypertension and with a normal glucose tolerance and serum lipid spectrum bopindolol in amounts of 0.5 to 2 mg/day in the course of 12 months. They revealed: a) bopindolol reduced effectively the blood pressure with a maximum decline already during the first month, b) It reduced significantly the heart rate, c) it did not cause deterioration of the glucose tolerance and did not interfere with the response of immunoreactive insulin (IRI) after a glucose load, d) it did not influence significantly the levels of lipoprotein cholesterol, total cholesterol, VLDL cholesterol, HDL cholesterol; within the reference range a slight increase of LDL cholesterol was recorded during the 12th month; e) it did not influence the concentration of triglycerides, apolipoprotein A1 and apolipoprotein B. PMID- 2573195 TI - [Mechanism of regulation of tyrosine metabolism in ethanol poisoning]. AB - Metabolism of tyrosine was impaired after chronic alcoholization of rats with 10% ethanol within 10 months. Within the first 3-4 months activation of tyrosine aminotransferase and a decrease in phenylalanine hydroxylase activity were found in liver tissue. Activity of tyrosine aminotransferase was not increased during the long-term alcohol intoxication. At the same time, activity of tyrosine aminotransferase was decreased within 5-6 months simultaneously with activation of phenylalanine hydroxylase. An increase in the alcohol dehydrogenase activity was also observed in rat liver tissue during the initial period of intoxication. The enzymatic activity was decreased beginning from the 3-4 months of the alcoholization and maintained at the low level. Hyperthermia augmented these alterations observed in chronic alcoholization of rats. PMID- 2573196 TI - [Analogy and differences in properties of soluble forms of guanylate cyclase of the heart and rat blood platelets]. AB - Activity of guanylate cyclase in rat thrombocytes exceeded the enzymatic activity in heart tissue 3.2- and 6.6-fold, if Mn2+ and Mg2+ were used as cofactors, respectively. Dithiothreitol (DTT) at concentrations 2 x 10(-5) M-2 x 10(-2) M activated guanylate cyclase both in rat heart and thrombocytes, while 2 x 10(-3) M of DTT exhibited the maximal stimulating effect: 3-fold in heart tissue and 4.5 fold in thrombocytes. Only slight 2-fold activation of guanylate cyclase was observed in myocardium in presence of 1 x 10(-4) M nitroprusside, whereas this effect was distinctly augmented up to 26-fold after preincubation of the enzyme with 1 x 10(-4) M of nitroprusside within 45 min at 4 degrees in presence of 2 x 10(-4) M DTT. The stimulating effect of nitroprusside was increased up to 52-fold after addition of 3 micrograms hemoglobin into the sample. Nitroprusside did not show any stimulating effect on the guanylate cyclase activity in rat thrombocytes under experimental conditions used. Possible causes of the phenomenon observed are discussed. PMID- 2573197 TI - [Decrease in the growth rate of Ehrlich's tumor and Pliss' lymphosarcoma with partial hepatectomy]. AB - In experiments with transplantable tumors, partial hepatectomy, Rhodiola rosea extracts or their combination were shown to inhibit the rate of Ehrlich's tumor and Pliss' lymphosarcoma as well as dissemination of the latter. These effects are to a certain extent attributed to production of humoral factors by the liver inhibiting clonogenic activity of tumor cells in vivo and in vitro. PMID- 2573198 TI - [A case of subdural abscess as a complication of suppurative meningitis]. AB - Subdural empyema is a rare intracranial complication of life-threatening importance. A case of this empyema was observed in a patient aged 29 years during purulent meningitis. The rarity of this disease may cause difficulties in differential diagnosis against more frequently occurring cerebral abscesses. Early diagnosis, in which computerized tomography is particularly helpful, makes possible early neurosurgical treatment which is of decisive significance for the survival. PMID- 2573199 TI - [Changes in the serum levels of lactate dehydrogenase, gamma- glutamyltransferase, glucose and alpha amino nitrogen in guinea pigs with experimental ascariasis]. AB - The studies were carried out on 90 male guinea pigs of which 75 were infected per os with a dose of ca. 5000 invasive eggs of Ascaris lumbricoides suis. At the 1st, 2nd, 4th, 7th and 20th day of infection blood was taken from experimental and control animals. The activity of LDH and its liver fraction (spectrophotometric method), GGTP (circle test method) was determined. The level of glucose (orthotoluidin method) and alpha amino nitrogen (ninhydrin method was determined. In the course of larval ascariasis an increase in the values of the studied parameters was noted, especially at the 4th and 7th day of infection. The changes in the activity of the studied enzymes evidence disturbances of the structure and function of the liver of infected animals. The increased level of glucose and alpha amino nitrogen is probably associated with a disturbance of basic energy metabolism of the host organism. PMID- 2573201 TI - Inhibition of lignocaine metabolism by beta-adrenoceptor antagonists in rat and human liver microsomes. AB - 1. The inhibition of lignocaine metabolism by beta-adrenoceptor antagonists (beta blockers) was investigated in rat and human liver microsomes. 2. Thirteen beta blockers (concn. 50 microM) incubated with substrate (4.27 microM) and rat liver microsomes, showed a strong linear correlation between percentage inhibition of lignocaine metabolism and the distribution coefficients of the beta-blockers (r2 = 0.842, P less than 0.001). Similar results for four beta-blockers were obtained using human liver microsomes. 3. In rat liver, which metabolizes lignocaine by aromatic hydroxylation and N-dealkylation, inhibition was selective for the former route. Human liver microsomes metabolize the drug mainly by N-dealkylation and inhibition of this pathway was observed. 4. Liver microsomes from rats treated orally with beta-blockers (0.34 nmol kg per day for 5 days) showed impaired metabolism of lignocaine and impaired formation of 3-hydroxy-lignocaine, despite the absence of significant residues of beta-blocker. 5. 14C-Propanolol was bound irreversibly to rat liver microsomal protein; binding accounted for 4.1 +/- 0.3% (n = 4) dose after 30 min incubation. Exclusion of co-factors and addition of glutathione (GSH, 1 mM) lowered binding by 96% and 70%, respectively. Propanolol inhibited lignocaine metabolism to the same extent in the presence or absence of GSH. The 14C-propanolol bound to liver microsomes from propranolol treated rats decreased in parallel with inhibition of lignocaine metabolism at 18 to 48 h after pretreatment. 6. These studies indicate at least two mechanisms for the inhibition of lignocaine metabolism by beta-blockers, namely, a 'lipid solubility hypothesis', where the effects may be related to the unchanged drug and a 'metabolite hypothesis', with the possible involvement of an irreversibly bound species. PMID- 2573203 TI - Influencing parenteral histamine2-receptor antagonist use. PMID- 2573202 TI - Phenothiazines and sudden infant death syndrome. PMID- 2573200 TI - Current concepts in inguinal hernia in infants and children. AB - Trends are changing in the management of infants and children with indirect inguinal hernias. Advances in neonatal intensive care have resulted in the survival of many small premature infants who have a high incidence of inguinal hernia. The rate of incarceration, strangulation, and gonadal infarction in these babies is twice that of the general pediatric age group. Respiratory immaturity, apnea, bradycardia, and associated neonatal conditions require special management at the time of hernia repair, usually performed just before discharge from the neonatal intensive care unit. New information concerning volume loss and depletion of germ cells beginning at 6 months of age in boys with undescended testes has stimulated the performance of orchiopexy when the patient is 1 year of age. More than 90% of boys with cryptorchid testes at the age of 1 year have an associated hernia that requires concomitant repair at the time of orchiopexy. The use of the peritoneal cavity for fluid absorptive purposes in hydrocephalus treated by venticuloperitoneal shunts or of peritoneal dialysis for renal failure and metabolic diseases such as hyperammonemia and lactic acidosis causes increased intraabdominal pressure and results in the appearance of a previously unrecognized hernia. Recognition of these and other conditions associated with a high incidence of hernial occurrence should allow early diagnosis and treatment before the development of complications. Most elective repairs of hernias are safely performed in the outpatient setting; however, some infants and children with concurrent illnesses are best managed in a "morning admissions" program, in which hospital admission occurs postoperatively. PMID- 2573204 TI - Contemporary issues in gastrointestinal diseases for clinical pharmacists. Proceedings of a symposium. April 22-24, 1988, Panama Beach, Florida. PMID- 2573205 TI - Comparison of the Parenteral histamine2-receptor antagonists. AB - The chemical structure, pharmacokinetic properties, and drug-drug interaction profiles of the parenterally available histamine2 (H2)-receptor antagonists were compared. Famotidine is a guanidinothiazole derivative, ranitidine contains an aminomethylfuran ring, and cimetidine has an imidazole ring. Data from the literature indicate that because of its chemical structure famotidine has a much greater potency and affinity for the H2-receptor and a notable lack of drug-drug interactions when compared with ranitidine and cimetidine. As a result, famotidine should be considered the H2-receptor antagonist of choice for critically ill patients who require gastric-acid suppression and at the same time are being treated with other drugs that depend on the cytochrome P-450 mixed function oxidase system for their metabolism and/or on renal tubular mechanisms for their excretion. PMID- 2573206 TI - Cost considerations of intravenously administered histamine2-receptor antagonists. AB - The cost of intravenously administered histamine2 (H2)-receptor antagonists to hospitalized patients is high. Costs can be expressed as either direct or indirect. Direct costs include drug cost, labor costs (pharmacy/nursing time), and the materials required for iv administration of these agents. Indirect costs include adverse effects, drug interactions, and allergic reactions. Due to the high percentage of total cost for labor and materials associated with the iv H2 receptor antagonists, a reduction in drug cost, although certainly desired, is unlikely to substantially reduce the amount charged the patient per intravenous dose administered or the daily cost of therapy. However, if less frequent dosing were required to achieve similar therapeutic effects, the daily therapy cost for iv H2-receptor antagonists could be substantially reduced. Assuming that cimetidine, ranitidine, and famotidine are equally effective and safe, our cost analyses at the University of Tennessee Medical Center/William F. Bowld Hospital indicate that famotidine administered q12h, regardless of the iv administration technique used, is the most cost-effective H2-receptor antagonist and is the drug of choice. PMID- 2573207 TI - Pharmacoeconomic aspects and formulary considerations related to histamine2 receptor antagonists. AB - Since the introduction of cimetidine in 1977, hospital expenditures for the histamine2-receptor antagonists have steadily increased, making them one of the most costly classes of therapeutic agents. As a result of cost considerations, we reviewed the available antiulcer agents in our hospital and evaluated the potential for therapeutic interchange within this group of agents. We found that by using famotidine as the principal agent in our hospital, drug cost savings of approximately $65,000 were realized. Additionally, avoiding the administration of approximately 13,000 parenteral doses with the selection of famotidine resulted in further reduction of drug administration costs. We conclude that the histamine2-receptor antagonists represent a cost-effective target for pharmacy cost-containment programs, without affecting patient therapeutic outcomes. PMID- 2573208 TI - Safety and acid-suppressant properties of histamine2-receptor antagonists for the prevention of stress-related mucosal damage in critical care patients. AB - Therapeutic intervention for the prevention of stress-related mucosal damage (SRMD) is currently directed at decreasing mucosal exposure to endogenous hydrochloric acid. We conducted a double-blind, parallel-group study in critically ill patients to investigate the comparative safety and acid suppressant properties of the intravenously administered H2-receptor antagonists cimetidine and famotidine. This is a preliminary analysis. Study patients received either famotidine 20 mg q12h or cimetidine 300 mg q6h or q12h. Cimetidine dosing intervals were based upon the degree of renal and hepatic dysfunction due to pharmacokinetic considerations. In critically ill patients with severe renal impairment, a pH of greater than or equal to 4.0 was maintained more consistently with famotidine 20 mg q12h compared with treatment with cimetidine. In terms of safety, a greater number of patients receiving cimetidine reported adverse clinical experiences than those receiving famotidine and a greater number of cimetidine patients had to be withdrawn from the study when compared with famotidine patients. In conclusion, preliminary results of this study suggest that famotidine has superior acid-suppressant properties and a better safety profile when used intravenously for the prevention of SRMD in critically ill patients. PMID- 2573209 TI - Use of continuous infusion of histamine2-receptor antagonists in critically ill patients. AB - Certain clinical situations require the use of a histamine2 (H2)-receptor antagonist to reduce gastric-acid volume and concentration or an antacid to act as a buffering agent. Presently, there are three H2-receptor antagonists available for iv use: cimetidine, ranitidine, and famotidine. Conventional therapy dictates that the H2-receptor antagonist be given by intermittent intravenous infusion, resulting in peaks and valleys of acid secretory control. Antacids, although capable of providing adequate gastric acidity control, must be administered frequently, often hourly, and thus require excessive nursing time. Presented here is a review of the rationale for the use of an H2-receptor antagonist by continuous infusion. PMID- 2573210 TI - [Classification of HIV-associated acquired immunologic deficiency syndrome]. AB - Regarding the staging of HIV infection, there are several classification systems, but neither of them has universally been accepted. We tested a new staging system with 4 parameters: HIV antibodies, CD4/CD8 ratio, absolute count of CD4 + lymphocytes, and absolute count of leukocytes. This classification indicates a regular progression of immunodeficiency, 294 blood specimens were used for immunological staging. The correlation between the immunological staging and the severity of the disease proved the usefulness of this classification system. PMID- 2573211 TI - Clinical pharmacology of multiple action compounds. AB - There are two principal divisions of the beta adrenoceptor blocking drugs in widespread clinical use, non-selective and selective agents, with or without intrinsic sympathomimetic activity. These properties confer differing pharmacological properties. A beta blocking drug with significant intrinsic sympathomimetic activity can be regarded as a multiple action drug. A third division of beta adrenoceptor blocking drugs is a newer development; these, which beside blocking the beta receptor, have an important peripheral vasodilator activity. Labetalol was the first drug of this group, prizidilol followed, but has been withdrawn because of toxicity. Several other agents are now under evaluation including bucindolol, carvedilol, dilevalol (one of the isomers of labetalol) and medroxalol. Celiprolol is an agent which is beta 1 selective and in addition has peripheral vasodilator activity. There are three mechanisms which have been described as responsible for peripheral vasodilation, alpha receptor blockade, beta 2 agonism, and a dilator action independent of either the alpha or beta receptors. Evidence for these various mechanisms is more readily obtainable from animal experiments, but some confirmatory evidence has been obtained in man. Inhibition of alpha stimulation has been found with labetalol, to a small degree with medroxalol and carvedilol, and has been suggested with celiprolol. Beta 2 mediated vasodilatation has been shown by celiprolol, dilevalol and medroxalol, and evidence of a vasodilatation independent of alpha or beta receptors has been obtained with celiprolol and carvedilol. More evidence is required to be sure of the exact contribution of each of these mechanisms. Combined action results in important haemodynamic differences from beta blockade, notably, peripheral resistance is reduced and less effect is seen on cardiac output.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573212 TI - Multiple action compounds in angina pectoris. AB - Carvedilol--a vasodilating beta-blocker has been shown in multicentre trials to be as effective as standard beta-blockers in improving exercise tolerance in patients with angina pectoris and it is more effective than nifedipine. The side effects are less than conventional beta-blockers and nifedipine. Carvedilol is an effective anti-anginal therapy and improves the quality of life. PMID- 2573213 TI - Effects of carvedilol in patients with impaired left ventricular function due to ischaemic heart disease. AB - In this single-blind, placebo-controlled trial, carvedilol, a nonselective beta blocking and vasodilating agent was studied in six patients with chronic stable angina. All patients had reproducible treadmill exercise time without medical treatment and developed chest pain in association with ST-segment depression (greater than 1 mm at J + 80 msec) on exercise. None had a history of rest or unstable angina or myocardial infarction within three months prior to the study. In all patients, anti-anginal medication except sublingual nitroglycerin was discontinued for 10 days. The patients entered an initial two week-phase of placebo. They then received carvedilol, 25 mg and then 50 mg twice daily for two weeks on each dose, followed by another two week-placebo-phase. Radionuclide ventriculography was performed at the end of each phase at rest and during maximal symptom-limited exercise. Bicycle ergometry was carried out in the supine position with incremental workloads. Exercise time and workload were recorded at the end of the first phase and imaging was performed at the same time and workload throughout the trial. Carvedilol produced a dose-related reduction in rest and exercise heart rate and blood pressure. Peak exercise ST-segment change was reduced by both doses of carvedilol, but this did not achieve a level of significance. After the first placebo phase all patients had abnormal left ventricular wall motion and resting ejection fraction (range: 35% to 45%). Four out of six patients had significant improvement in wall motion abnormalities, in two patients there was no change, and none developed a deterioration in abnormal wall motion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573214 TI - Effects of carvedilol on left ventricular function and arrhythmias during repeated short-time myocardial ischemia in experimental pigs. AB - The effects of carvedilol, a new vasodilating beta-blocking drug, were studied in experimental pigs during short-term acute myocardial ischemia. In 21 anesthetized pigs 0.01, 0.03, and 0.1 mg/kg b.w. carvedilol was studied during repeated 2-min distal LAD-occlusions and a 60-min-period of ischemia. Left ventricular volume was continuously measured by the impedance catheter method. Intravenous administration of 0.01 mg/kg carvedilol resulted in a significant decrease of heart rate (-13%), dp/dtmax (-32%), and ejection fraction (-9%), slight changes of systolic pressure (-3%), and an increase of vascular resistance (+24%), indicating a beta-blocker effect without vasodilation-while the first vasodilatory effect was found at a dose of 0.1 mg/kg. During ischemia carvedilol had no influence on the time-course or the extent of systolic bulging of the ischemic myocardium, but the ischemia-induced decrease of left ventricular ejection fraction was diminished. Both during short-term ischemia, as well as during the 60-min-ischemia-period carvedilol significantly reduced ventricular premature beats. During the 60-min-ischemia-period activation delay measured from local DC-electrograms of the ischemic myocardium, as well as the occurrence of activation block were not altered by carvedilol, as was the incidence of ventricular fibrillation (69%). We conclude that at low dosages the beta-blocking effect of carvedilol exceeds the vasodilating properties. This may also hold true in patients with cardiac failure; they are more sensitive to beta-blocking drugs. During ischemia carvedilol slightly reduces the ischemia-dependent decrease of global ventricular function and it has an antiarrhythmic effect. Therefore, it may be protective in patients with acute myocardial infarction. PMID- 2573215 TI - Serospecific protection of mice against intranasal infection with Bordetella pertussis. AB - The ability of purified serospecific agglutinogens from Bordetella pertussis to protect mice against intranasal infection has been examined. Immunization with agglutinogen 2 protected mice against infection with 1.2.0 or 1.2.3 serotypes of B. pertussis, whereas immunization with agglutinogen 3 protected mice against infection with all serotypes. More importantly immunization with serospecific agglutinogen resulted in immune selection so that organisms recovered following infection did not express the immunizing antigen. The results are consistent with the suggestions that protection of children with whole cell pertussis vaccine is to some extent serospecific and that agglutinogens should be considered as constituents of acellular pertussis vaccines. PMID- 2573216 TI - Significance of toxin-coregulated pili as protective antigens of Vibrio cholerae in the infant mouse model. AB - The infant mouse cholera model has been used to evaluate the relative importance of toxin-coregulated pili (TCP) as protective antigens of Vibrio cholerae 01. Electron microscopic and immunoblotting analyses revealed that, under the cultural conditions examined, TCP were only expressed by strains of classical biotype. Antibodies to TCP were sufficient to confer protection against two such strains, and were more efficient if the challenge vibrios were cultured for TCP expression. In contrast, such antibodies did not protect mice against challenge with any of four strains of El Tor biotype. Since two of the latter have previously been shown to possess non-lipopolysaccharide protective antigens, these results suggest that TCP are not the only such antigen in this model. PMID- 2573217 TI - [Glucocorticoid receptors as targets for pharmacologic action]. AB - Experimental data demonstrate the influence of certain drugs on the interaction of Type II and III glucocorticoid receptors of the liver cytosol and labelled natural and synthetic glucocorticoids. Preparations of the phenothiazine (aminazine, tizercine) and pyrazolone (amidopyrine, analginum) series and non steroid anti-inflammatory drugs (acetylsalicylic acid, sodium salicylate) exert a significant effect on the glucocorticoid-receptor interplay. Certain drugs (tizercine, analginum) have an anti-stress effect by lowering the level of Type II glucocorticoid receptors of liver cytosol and corticosterone concentration in the blood plasma of rats. The phenothiazine and no-spa derivatives inhibit the matrix activity of the thymocyte DNA in adrenalectomized rats. Pyracetam, caffeine, adrenaline, noradrenaline, verapamil, digoxin, and streptomycin do not influence the thymocyte DNA matrix activity in the presence of triamcinolone acetonide. Based on the literature data and original research findings, the problem of glucocorticoid receptors employment as targets for medication to control the glucocorticoid effect and stress is discussed. PMID- 2573218 TI - [The beta-blocker effect of the blood serum in patients with psoriasis]. AB - A short-term cultivation of human skin combined with histoautoradiography have demonstrated that the blood serum of psoriasis patients is characterized by an in vitro effect similar to the beta-blocker effect, i.e. eliminates the epidermal chalone action, thus enhancing the epidermal cell proliferation in foci of skin involvement in psoriasis. PMID- 2573220 TI - [The role of cGMP in extinguishing the reactions of identified neurons in the edible snail to acetylcholine]. AB - Possible role of cGMP is studied in control of extinction of snail neurones RPa4, RPa3 and LPa3 reactions to acetylcholine (ACh), applied rhythmically to neurone soma by means of microiontophoresis. It is shown that guanylate cyclase activators which raise the cGMP level in the cell--Na nitroprusside and Na azide (5,10(-4)-10(-3) mol/l)--intensify at extracellular application the extinction of inward transmembrane current and membrane depolarization in response to ACh. Suggestion is made about participation of cGMP-dependent phosphorylation of membrane proteins in control of the development rate, depth and duration of neurone cholinoreceptors short-term plasticity. PMID- 2573219 TI - [Temporal specificity in stimulus action during the formation of associative ultrastructural restructurings in cerebral cortex neurons]. AB - By a method of electronic microscopy morphological characteristics were studied of various components of neurones synapses in the sensorimotor cortical zone of rats at different variants of combined and noncombined repeated microiontophoretic presentation of glutamate and acetylcholine. Significant dependence was found of the character and rate of reorganizations of postsynaptic thickness (PSTh), width of synaptic cleft and length of synapses active zone on temporal relations in transmitters action: significant changes of PSTh thickness appeared only in conditions of combined presentations of stimuli (neurotransmitters); maximum thickening of PsTh was caused by the combined action of glutamate and acetylcholine with 3 s delay of the latter. The hypothesis is suggested that temporal specificity during integration of associated signals action to neurones is determined by kinetics of interacting biochemical regulating mechanisms. PMID- 2573221 TI - [Changes in the brain electrical activity under the influence of beta adrenoreceptor agonists and antagonists in rats during free behavior]. PMID- 2573222 TI - [Comparison of Cabot/Nesbit orchidopexy with Petrivalsky/Schoemaker orchidopexy]. AB - Two groups of each 50 children with maldescensus testis treated by two different techniques of orchiopexy are compared together. Two to five years after operation the localization and size of the testes are recorded. In the group of children with orchiopexy according to Cabot/Nesbit the failure rate was 12%, in contrast to only 6% failures and larger testes in the Petrivalsky/Shoemaker group. Therefore, the technique of Petrivalsky/Shoemaker is recommended, especially in small children. PMID- 2573223 TI - [ Ulcerative colitis and pregnancy]. AB - Ulcerative colitis is a disease confined to the colon in an age from 20 to 40 years mainly. Fertility and pregnancy are not disturbed generally. The influence of pregnancy on the course of the colitis is not foreseeable, however. The therapy with Salazosulfapyridine is possible in pregnancy, too. PMID- 2573224 TI - The prevalence of Hantaan viruses among Rattus norvegicus in Tokyo in the 1960's. AB - During the 1960's, 15 out of 89 rats (16.8%) captured in the port area and 6 out of 59 rats (10.2%) in the suburb of Tokyo were seropositive against Hantaan viruses. PMID- 2573225 TI - Intracellular localization of tyrosine hydroxylase immunoreactivity in the rat adrenal chromaffin and pheochromocytoma cells. AB - The localization of tyrosine hydroxylase (TH) immunoreactivity in rat adrenal chromaffin and pheochromocytoma (PC12) cells was investigated by immunoelectron microscopy using monoclonal and polyclonal antisera against TH purified from rat adrenal medulla. Strong TH immunoreactivity was found uniformly in the granules of the adrenaline cells; the immunoreactivity was visible mainly within the periphery, but not in the clear space of the granules of the noradrenaline cells. In the PC12 cells, strong TH immunoreactivity was also observed uniformly in the granules. In addition, TH immunoreactivity was seen in the cytoplasm, the ribosomes attached to the endoplasmic reticulum and the free ribosomes of both the rat adrenal chromaffin and PC12 cells. These results suggest that TH may be localized in the granules, cytoplasm and ribosomes of rat adrenal chromaffin and PC12 cells. PMID- 2573226 TI - Tyrosine-hydroxylase-immunoreactive nerve fibers in the separated capsule of the rat adrenal gland. AB - The present study applied the separated adrenal capsules of rats for wholemount immunocytochemistry and used tyrosine hydroxylase (TH) antibody as a marker for catecholamines. TH-immunoreactive nerve bundles without varicosities and fibers with varicosities were seen to run along or to encircle blood vessels entering the adrenal capsule from the outside, and then to run along a network of blood vessels in the intracapsular region. Also, the TH-immunoreactive nerve bundles and fibers were found to run along blood vessels in the subcapsular region. Some TH-immunoreactive nerve fibers and bundles with varicosities, unassociated with the blood vessels, were seen in the subcapsular region. In this region, TH immunoreactive nerve fibers with varicosities were often seen to be closely associated with the cortical cells. Some TH-immunoreactive nerve fibers without varicosities were visible within the splanchnic nerve in the subcapsular region. The present study suggests that numerous catecholaminergic nerve fibers are associated with blood vessels forming a network in the superficial region of the rat adrenal gland. PMID- 2573227 TI - Peptidergic and catecholaminergic fibers in the human corneal epithelium. An immunohistochemical and electron microscopic study. AB - Innervation of the clinically normal human corneal epithelium was investigated utilizing immunohistochemical and electron microscopic techniques. All corneal epithelial sheets examined demonstrated neuron specific enolase (NSE: a non specific marker for neural elements), calcitonin gene-related peptide (CGRP: a putative marker for sensory fibers), and tyrosine hydroxylase (TH: a marker for catecholaminergic nerves) immunoreactive fibers. NSE, CGRP, and TH fibers formed a dense basal epithelial plexus. The CGrp fibers tended to have beaded profiles, while TH fibers were smooth. Numerous free nerve endings originating from the basal epithelial plexus og NSE and CGRP fibers terminated throughout the thickness of epithelium. The densities of fibers in the basal epithelial nerve plexus were: NSE greater than CGRP greater than TH. Transmission electron microscopy demonstrated two types of epithelial nerve fibers, one containing large dense-core vesicles and another small dense-core vesicles. Both types contained clear vesicles. These large and small dense-core vesicle fibers appeared to correspond to the CGRP and TH immunoreactive fibers, respectively. These results provide morphological baseline data on the normal sensory and sympathetic corneal epithelial innervation. PMID- 2573228 TI - Less pain with epidural morphine after knee arthroplasty. AB - Twenty-two patients were randomly allocated to systemic opioids or epidural morphine the first 10 days after total knee arthroplasty. Pain was recorded daily in a visual analogue scale, and knee motion was measured on Day 10. Pain was lower in the epidural group, with no difference in knee flexion or range of motion. PMID- 2573229 TI - High-voltage electron microscopic observations on the suprastructure of the macula utricle. AB - Thick sections of the suprastructure of the utricular macula of the guinea pig were observed by high-voltage electron microscope. Whole views and the relationship of the otoconia, the otoconial membrane and the sensory ciliary bundles became very clear. In addition, stereo micrographs were even more helpful for simultaneously recognizing this three-dimensional relationship. PMID- 2573230 TI - Molecular genetics of Turner's syndrome. AB - Recombinant DNA technology now allows an analysis of sex chromosomal abnormalities at the molecular level. X chromosomes differ in respect of their pattern of cutting sites for particular restriction enzymes; these differences can be used to determine which X chromosomes or which part of an X chromosome has been inherited from a parent. Furthermore, these techniques can be used to define the presence or absence of particular regions of the X chromosomes with a higher level of resolution than it is possible to achieve using light microscopy. Thirty eight families in which there was a child or fetus with Turner's syndrome were studied using a series of DNA probes that detect differences (restriction fragment length polymorphisms) among X chromosomes. Analysis of the origin of the normal X chromosome was possible in 27 families. In 14 families with 45,X the observed X was maternal in each, whereas in 13 children with other karyotypes (46,X,i(Xq); 45,X/46,X,i(Xq); 45,X/46,XX; 45,X/46,X,r(X)) the origin of the normal X was paternal in six and maternal in seven patients. In two other families, mosaicism, which was unsuspected at chromosomal analysis, was revealed. DNA probes studied in the remaining nine families were uninformative. These results suggest that different pathogenic mechanisms operate for pure 45,X when compared with the other karyotypes associated with Turner's syndrome. The presence of unsuspected mosaicism in some of these families has clinical implications in relation to prognosis and management. PMID- 2573231 TI - [Application of Pirkle-type chiral column to the resolution of enantiomeric drugs: resolution and identification of alpha-naphthyl isocyanate derivatives of alprenolol and some other beta-blockers]. AB - alpha-Naphthyl isocyanate (NI) as a derivatizing agent to separate several beta blockers was described. This procedure is simple, rapid and gives good resolution. Experiments with NI derivatives of alprenolol showed that the isocyanate group reacted with the amino group on the analyte to form urea under mild conditions and with the hydroxyl group to form carbamate under relatively vigorous conditions. The results also indicated that the derivatized reactions is 1:1 between NT and analytes. PMID- 2573232 TI - Transmitter- and voltage-activated ionic channels in astrocytes: possible role in extracellular ionic homeostasis. PMID- 2573233 TI - Hypothalamic neurosecretory systems and their messenger molecules. AB - The distribution and cellular localization of several transmitters, transmitter synthesizing enzymes and peptides in the arcuate, paraventricular and supraoptic nuclei are described with special focus on coexistence of messenger molecules. The role of these compounds in various neuroendocrine events is discussed. PMID- 2573236 TI - Melperone-an atypical neuroleptic. Proceedings of a workshop. Malmo, Sweden, October 8, 1986. PMID- 2573234 TI - A double-blind study of adjunctive carbamazepine versus placebo on excited states of schizophrenic and schizoaffective disorders. AB - A multi-institutional double-blind study comparing the therapeutic effect of adjunctive carbamazepine and placebo with standard neuroleptic treatment was performed on 162 patients with DSM-III diagnosis of schizophrenic (n = 127) or schizoaffective disorders (n = 35) who had excited states or aggressive/violent behavior that responded unsatisfactorily to neuroleptic treatment. The patients participated in a 4-week trial of carbamazepine plus neuroleptics (n = 82) or placebo plus neuroleptics (n = 80). The sum of patients with marked and moderate improvement was modestly larger in the carbamazepine group (48 vs. 30%, P less than 0.05). There was no significant difference between the carbamazepine and placebo groups in the changes of total BPRS scores, although the carbamazepine group showed more improvement on the items suspiciousness, uncooperativeness and excitement. The results suggest that carbamazepine, when used in combination with neuroleptics, is a useful drug for the treatment of excited states of patients with schizophrenic and schizoaffective disorders. PMID- 2573235 TI - Benzodiazepine withdrawal and rebound insomnia. PMID- 2573237 TI - Approaches for finding new types of antipsychotic compounds. AB - There is strong evidence that pharmacological interaction with central dopamine transmission at several levels may induce antipsychotic effects in psychiatric patients. All the pharmacological principles interacting with other systems have so far not been established with regard to antipsychotic potential. Besides the dopamine system pharmacological manipulation of central serotonergic, gabaergic and noradrenergic mechanisms may also be of therapeutic value in schizophrenia. The interaction of peptidergic mechanisms and central dopaminergic mechanisms is another avenue for further exploration regarding to pathophysiology of psychosis and its treatment. Elucidation of the heterogeneity and molecular structure of central dopamine receptors seems the most rational approach for creating new dimensions for further development of drug design for pharmacotherapy of psychosis. The detailed mapping of afferent and efferent systems coupled to central dopamine neurons is also an approach that may give guide-lines for finding new types of antipsychotic drugs. PMID- 2573238 TI - Melperone and clozapine: neuroendocrine effects of atypical neuroleptic drugs. AB - The effects of atypical neuroleptics within the neuroendocrine axis of rodents can be distinguished from those of typical neuroleptics by the production of: 1) a shortlived increase in serum PRL concentrations, 2) an acute increase in the activity of TIDA neurons, and 3) a marked increase in serum corticosterone concentrations. It is of interest to speculate that the pharmacological properties of atypical neuroleptics which mediate the unique neuroendocrine responses are of relevance to an understanding of the mechanisms which underlie the clinical profile of these antipsychotic agents. PMID- 2573239 TI - Clinical melperone treatment blocks D2-dopamine receptors in the human brain as determined by PET. AB - Positron emission tomography and 11-C-labelled raclopride was used to determine central D2-dopamine receptor occupancy in three melperone treated patients. Treatment with melperone in daily doses of 250 and 300 mg for 3 to 6 weeks, resulted in a receptor occupancy above 70%. Thus, clinical doses of melperone as we previously demonstrated for several classical neuroleptics cause a substantial D2-dopamine receptor blockade in the human brain in vivo. PMID- 2573240 TI - A clinical comparison of melperone and placebo in schizophrenic women on a milieu therapeutic ward. AB - Twenty schizophrenic women participated in a double-blind comparison of melperone (300 mg daily) and placebo lasting 4 weeks. All the patients received milieu treatment. Two patients in the melperone and four in the placebo group were removed from the study due to unsatisfactory therapeutic response. In the melperone group, there were significant reductions in psychotic morbidity. In the placebo group, there were occasional significant reductions of psychotic morbidity during but not at the end of treatment. The melperone-treated patients exhibited significantly lower morbidity scores than the placebo group after treatment for two and four weeks. The melperone group demonstrated significantly more adverse reactions than placebo-treated patients at the end of the study. The results support the view that melperone exerts an antipsychotic effect. The study is consistent with the opinion that milieu therapy should be combined with neuroleptics to achieve an optimal result in the treatment of an acute phase of schizophrenia. PMID- 2573241 TI - Chronic melperone administration does not enhance oral movements in rats. AB - Melperone and haloperidol were compared in a rat model for tardive dyskinesia. Drugs were given chronically for 12 months with the drinking water and the frequency of oral movements was measured at monthly intervals. Haloperidol (0.2 and 0.4 mg/kg/d) produced an increase in vacuous chewing movements, whereas melperone (2.4, 5.1 and 11.0 mg/kg/d) did not differ from untreated controls. PMID- 2573242 TI - Pharmacological data of the atypical neuroleptic compound melperone (Buronil). AB - Based on the non-cataleptic properties, the weak affinity for D2 receptors and the inability to induce DA receptor supersensitivity after both acute and repeated administration, melperone may be characterized as an atypical neuroleptic drug. This indicates a weak effect of melperone on striatal DA neurotransmission. On the other hand melperone is potent in blocking amphetamine induced locomotion, exploratory behaviour, L-Dopa induced jumping and aggression suggesting a limbic mode of action. The effect on limbic DA neurotransmission together with an antiadrenergic and 5-HT modulating action may explain the antipsychotic effect and the low incidence of extrapyramidal side effects observed during treatment with melperone. PMID- 2573243 TI - The neuro-endocrine ambiguity of sympathoadrenal cells. AB - This introductory article presents a survey of the principle cellular constituents of the sympathoadrenal section in the peripheral autonomic nervous system, their development and plasticity and factors that govern the expression of particular morphologic and transmitter phenotypes. The article focuses on results obtained in cell culture studies with isolated chromaffin cells that have permitted the analysis of molecular signals possibly serving as environmental cues during the development of sympathoadrenal cells. PMID- 2573244 TI - Catecholaminergic cells and support cell precursors in neural crest cultures differentially express nerve growth factor receptors. AB - Long-term neural crest cultures grown in the continuous absence of exogenous nerve growth factor (NGF) contain a subpopulation of cells with NGF receptors exclusively of the low affinity subtype (Kd of approximately 3.2 nM). The current studies combined immunocytochemistry, using GIN1 (a support cell marker) or tyrosine hydroxylase antibodies, with radioautography following exposure to iodinated nerve growth factor (125I-NGF). The majority of cells specifically binding 125I-NGF were found to be immunoreactive for GIN1, indicating that the primary cell phenotype expressing receptors for NGF appear to be support cell precursors, at least under these conditions. These cells are likely to be responsive to and/or dependent upon NGF; the nature of this response or dependency remains to be determined. Some cells exhibiting silver grains were not immunoreactive for GIN1, suggesting that other cell phenotypes in neural crest cultures also have NGF receptors. In addition, some neural crest cells were found that stained with GIN1 and lacked 125I-NGF binding. Tyrosine hydroxylase-like immunoreactive cells apparently did not bind 125I-NGF under these culture conditions. Catecholaminergic sympathetic and sensory neurons from embryonic ganglia, derived from the neural crest, express both the high and low affinity forms of the NGF receptor. In order to determine whether the microenvironment played a role in the type of catecholaminergic cells appearing in culture, neural crest cells were grown in the continuous presence of exogenous NGF. Under these conditions, many tyrosine hydroxylase-like immunoreactive cells were found that specifically bound 125I-NGF. In addition, silver grains were still detected on these cells following a chase with nonradioactive NGF, designed to eliminate 125I NGF bound to low affinity sites. Therefore, the catecholaminergic cells possess both the low and high affinity forms of the receptor. NGF's ability to modulate tyrosine hydroxylase activity, as it does in mature catecholaminergic neurons, was tested in this system. Surprisingly, there was no statistically significant difference in tyrosine hydroxylase activity in cultures grown in the absence or presence of exogenous NGF. This raises the possibility that embryonic catecholaminergic cells are unable to respond to NGF in this specific way, even though the receptors for the factor are present. PMID- 2573245 TI - Co-localization and plasticity of transmitters in peripheral autonomic and sensory neurons. AB - Immunohistochemical studies have shown that most peripheral autonomic and sensory ganglia are heterogeneous, consisting of several populations of neurons which can be distinguished by their content of peptide and non-peptide transmitters, and transmitter-associated enzymes. Many neurons contain several different potential transmitters, especially neuropeptides. Some neuropeptides have been localized in more than one population of autonomic and sensory neurons. However, the peptide often occurs together with a distinctive combination of additional transmitters in each neuronal class. The precise combination of transmitters found in any individual neuron is highly correlated with the peripheral target of the neuron. This indicates that immunohistochemically defined neuronal populations represent distinct functional classes of neurons. In an increasing number of cases, many of the potential transmitters contained in a particular neuron have been shown to be released from the nerve terminals, and to contribute to presynaptic or postsynaptic effects of nerve activation. Despite this association between the combination of potential transmitters contained in a neuron, and the function of the neuron, not all transmitters or transmitter-associated enzymes are expressed equally at all times in the life of a neuron: the levels of some substances change dramatically during development; some are detected only after experimental alteration of the environment of the developing or mature neurons. Taken together, these results indicate that, during development, pathway-specific information influences the differentiation of peripheral autonomic and sensory neurons. Furthermore, the expression of neuropeptides and transmitter-associated enzymes in a particular neuron appears to be under continuous regulation. These phenomena demonstrate the complexity and precision involved in development and maintenance of the peripheral autonomic and sensory nervous systems. PMID- 2573246 TI - Use of somatic cell mutants to study the signal transduction function of the T cell antigen receptor. PMID- 2573247 TI - [Plate osteosynthesis of forearm fractures in adults and adolescents. Long-term results and problems]. AB - Based on the results reported in the literature, the method applied by us in the management of forearm shaft fractures is presented and explained. The advantages and remaining problems of the method are discussed on the basis of the results obtained in the follow-up of 45 ulna-, radius- or forearm shaft fractures treated by means of DC-plate osteosynthesis. The procedure is complication-free and has a low pseudarthrosis rate. To avoid refractures, the material should not be removed earlier than one and a half year following osteosynthesis. Axis shifts of more than 10 degrees and differences in length of more than 2 mm should be avoided and should be corrected in working patients. PMID- 2573248 TI - [Surgical treatment of aseptic forearm shaft pseudarthrosis]. AB - Non union of a lower arm fracture (without infection) is the result of inadequate immobilization or of too extensive devastation of the bone. Within 1975 to 1985 in the accident-hospital of Tubingen we saw 131 pseudarthroses of the ulnar or radius shaft in 111 patients. Among these a hypertrophic pseudarthrosis is mostly seen after conservative treatment or operative treatment with intramedullary nailing. Atrophic pseudarthroses or defect pseudarthroses mostly occur after open fractures, comminuted fractures or insufficient plate osteosynthesis. The operative treatment was regularly a plating of radius and/or ulna (mostly with the 3.5 mm AO-DC-plate) combined with correction of axis or rotation deformity. Depending on the type of the pseudarthrosis, decortication was also performed as well as an autogenous bone-grafting or an interposition of a cortical-cancellous block. All but one of the pseudarthroses each of the ulna and radius, healed completely. Most patients had a reduction of pain, improvement of mobility of the elbow or wrist joint, especially an improvement of supination/pronation, regarded as an index of correction of a malrotation. Postoperative physiotherapy has to be most careful; the best physiotherapy is the everyday active use of the arm by the patient himself. PMID- 2573249 TI - [Bone contact of miniscrews with and without a pretapped thread]. AB - Better contact between screw and bone when using 2 mm AO-Miniscrews in the human midface is achieved if the thread is not pre-tapped. This could be demonstrated on metacrylate sections. In most implants with pre-tapped threads the bone does not fill out the space between the threaded core and the pretapped hole. The average measured distance between screw and bone is 110 microns. The contact surface between bone and screw is considerably greater than with pre-tapped threads. PMID- 2573250 TI - [Resection of the coracoacromial ligament. A simple surgical procedure for decompression of the subacromial space]. AB - Resection of the coracoacromial ligament is a simple method for decompression of the subacromial space. The operation can be performed under arthroscopic control or by means of a small incision above the ligament. 61 patients were operated on by this method during the period between July 1987 and August 1988. 37 patients were questioned regarding the results. In 40% of these there was an alleviation of pain, 48% had an improved motility and 67% would again agree to the operation that appeared indicated (in the patients referred for surgery) both in the impingement syndrome and in antiquated ruptures of the rotator cuffs. PMID- 2573251 TI - [Cause, prognosis and therapy of tibial pseudarthroses]. AB - A report on surgical treatment of 75 pseudarthroses of the tibia, 64 cases of which could be followed up for assessing the treatment results. The majority of these were pseudarthroses due to infection or associated with atrophy, that is to say, cases that were difficult to treat. Stabilization was usually effected by osteosynthesis with autocompression plates; in the majority of cases autologous bone grafts were performed additionally. In 82.8% of the cases cure of the pseudarthrosis was achieved, in 68.8% already by first surgery. 7.8% of the patients refused further surgical treatment. 9.4% are still under treatment at the time of writing. PMID- 2573252 TI - [Bristow surgical treatment of shoulder dislocation]. AB - 14 shoulders with recurrent anterior dislocation were treated with a modified Bristow procedure and had a 38 to 66 months follow up after surgery. Postoperatively no patient had recurrent anterior dislocation. 4 patients had an average loss of external rotation of 10-15 degrees, one patient had a non-union of the transplanted coracoid. None of the patients was disappointed by the results. PMID- 2573254 TI - [Menstruation and postoperative complications. A review of the literature]. AB - Because of a Toxic Shock Syndrome following an urgent knee operation during the premenstrual period we reviewed the available literature. There is no indication that the physiological process in woundhealing is influenced by menstruation. Increased woundinfection after operations during the perimenstruum were--except for inner female genitals--not observed. But more and more TSS is described after any kind of operation, so traumatologists should be well acquainted with the symptoms. PMID- 2573253 TI - [Intrathoracic hemorrhage of the intra-abdominal organs following diaphragmatic rupture. A rare cause of hemothorax]. AB - After posttraumatic diaphragm ruptures in some cases life threatening haemorrhages from intraabdominal organs (especially from the spleen) into the thorax were found. In general, the diagnosis cannot be established by peritoneal lavage or abdominal sonography. And in case of emergency, there is often not enough time to obtain a computed tomography. It is therefore extremely important for the patient's prognosis that the surgeon initially performing treatment be aware of this cause of bleeding. Correct interpretation of the chest x-ray, for instance in the case of stomach air in a left basal shadow above the diaphragm, can confirm the suspicion, and the urgently required operation can be performed in time. PMID- 2573255 TI - [Secondary wire migration following percutaneous bore wire fixation of acromioclavicular dislocation]. PMID- 2573257 TI - [Pulsed lasers in the treatment of urinary calculi]. AB - Intracorporeal lithotripsy of urinary calculi by pulsed dye laser recently enriched the urologist's therapeutic arsenal. We recall the in vitro studies and animal experimentations which demonstrated the feasibility of laser lithotripsy and the absence of harmful tissue damage. The ideal indications for intracorporeal lithotripsy are ureteral stones accessible by rigid and, more especially, flexible ureteroscopy. The results of the first published series using this technique are very encouraging. PMID- 2573256 TI - The immunopathogenesis of HIV infection. PMID- 2573258 TI - [Testicular tumor following orchiopexy: a case report]. AB - Herein we report a 26-year-old patient in whom seminoma developed in the testicle which had been fixed 9 years earlier. Thirty five of such cases in the Japanese literature are reviewed. PMID- 2573260 TI - [Clinical relevance of determining lymphocyte subpopulations in peripheral blood of patients with rheumatoid arthritis]. AB - Blood lymphocytes were determined from 21 patients with seropositive rheumatoid arthritis (RA) in a cross-sectional study with 5 monoclonal antibodies (MAb) of the BL-series. 7 patients were examined with the same MAb over a time period of 14 weeks before and after a prednisolone-pulse therapy. The cross-sectional study showed a significant higher percentage of Ia4+ T-helper/inducer-lymphocytes in RA patients compared to control persons. The value of Ia4+ T-lymphocytes didn't correlate to the clinical activity of the RA. No significant differences were observed in the helper/suppressor T-lymphocyte ratio. The lymphocyte subpopulations were not significantly different immediately before and after a prednisolone-pulse therapy. The percentage of B-lymphocytes was significantly higher in comparison to the controls 24 h after prednisolone-pulse therapy. An immunomonitoring of the clinical activity and the therapy of RA is not possible with the used MAb. PMID- 2573259 TI - Pirenzepine treatment in urticaria-angioedema syndrome caused by adverse reactions to foods. AB - An increased gastroenteric mucosal permeability is generally considered a pathophysiological mechanism in the urticaria-angioedema syndrome caused by adverse reactions to foods. Since pirenzepine, an antimuscarinic receptor drug, exerts a cytoprotective activity on digestive mucosa, the authors evaluated the clinical efficacy of pirenzepine and terfenadine (antihistamine), alone or associated, in the treatment of patients with urticaria-angioedema syndrome due to food allergy. Furthermore, additional endoscopy and biopsy studies were performed in order to provide experimental evidence about the cytoprotective activity of this treatment. The results of the present investigation confirm the clinical efficacy, with improvement of histological parameters, of pirenzepine treatment in adverse reactions to foods, as previously demonstrated by our group, and suggest further investigations on the functional mucosal impairment hypothesized in this pathological condition. PMID- 2573261 TI - Drug abuse: a biopsychiatric model. AB - The effects of drug abuse are caused by the stimulation or inhibition of different neurotransmitters, chiefly gamma-aminobutyric acid, acetylcholine, norepinephrine, dopamine, serotonin and beta-endorphin. The biopsychiatric model focuses on neurotransmitter activity to diagnose and treat overdose and addiction. This model explains how different drugs exert their effects and provides a rationale for specific pharmacologic intervention in the drug-abusing patient. PMID- 2573262 TI - Initial drug therapy for hypertensive patients with hyperlipidemia. AB - Hypertension and hyperlipidemia are cardiovascular risk factors that commonly coexist. Studies have indicated that it is important to control both risk factors to achieve significant reductions in morbidity and mortality. Recent debate has focused upon whether traditional step I antihypertensive agents can substantially lower these risks because of their effects on plasma lipids. This debate continues to be unresolved. However, for the patient with elevated lipid levels, diuretics and beta-blockers may make the management of the lipid disorder more difficult. Therefore it may be desirable to select alternative step I antihypertensive agents that will not interfere with the therapy for hyperlipidemia. Alternative step I agents include alpha 1-blockers, ACE inhibitors, and calcium channel blockers. These agents either have no effect on plasma lipids or they improve the lipid profile. Generally, these drugs are well tolerated and provide good alternatives for patients with hyperlipidemias. The initial drug of choice can be chosen depending upon other patient variables such as age, race, or concomitant diseases. PMID- 2573263 TI - Therapeutic balloon occlusion and pharmacologic therapy of a right-to-left atrial shunt produced by right ventricular infarction. PMID- 2573264 TI - Exercise testing and training with beta-adrenergic blockade: role of the drug washout period in "unmasking" a training effect. AB - To determine whether or not a training effect can be achieved with beta adrenergic blockade and whether there is a difference between selective and nonselective therapy, we recruited 40 healthy subjects (16 women, 24 men) to participate in a 9-week exercise training program. After a baseline exercise treadmill test, subjects were randomized to oral therapy groups of atenolol, 50 mg daily (AT 50), atenolol, 100 mg daily (AT 100), propranolol, 80 mg twice a day (Prop), or placebo. Repeat exercise tests were performed at week 1, week 8, and at week 9, with week 8 to 9 being a 1-week drug-free washout period. At week 8, maximal oxygen consumption (Max VO2), when compared with baseline levels, was increased slightly in AT 50 (4.2%) and Prop (2.4%), decreased in AT 100 (5.3%), and increased significantly in the placebo group (12.7%). After washout, Max VO2 increased significantly compared with baseline in AT 50, AT 100, and Prop (9.8%, 10.8%, and 9.8%, respectively). We conclude that there is no significant difference between selective and nonselective beta-blockade therapy in the development of a training effect. This effect, however, may not become apparent until the drug is withdrawn. PMID- 2573265 TI - The safety of amlodipine. AB - The safety profile of amlodipine was assessed from the pooled data base of clinical research studies. This data base included 4227 subjects, 2495 of whom received amlodipine (including 2189 who received multiple-dose amlodipine); the remainder received comparative agents (placebo 1213; active comparatives 519). Amlodipine treatment was associated with a slightly higher incidence of side effects compared with placebo, but most of this difference was the result of edema, which was usually well tolerated. When compared with the beta-blockers atenolol and nadolol, amlodipine had a favorable safety profile. In particular, the incidence of severe side effects in patients receiving amlodipine was approximately half that reported for patients receiving beta-blockers. The data base comparing different calcium antagonists was small; in a study versus verapamil, edema was more common in patients receiving amlodipine, but constipation was more common in patients receiving verapamil. In a study versus diltiazem, both amlodipine and diltiazem were similarly well tolerated. Amlodipine was not associated with the deleterious effects of serum creatinine, urate, and fasting glucose, which was caused by hydrochlorothiazide, and in contrast to hydrochlorothiazide and nadolol, amlodipine was not associated with unfavorable changes in serum cholesterol and serum triglyceride levels. Amlodipine was well tolerated by elderly patients and is not contraindicated in patients with conduction abnormalities. Dosage modifications are unnecessary in renal impairment, but the dosage regimen for patients with hepatic impairment is not yet established. Amlodipine is an antihypertensive and antiischemic agent that has the combined advantages of a good safety profile with once-daily dosage and a smooth onset and long duration of action. PMID- 2573266 TI - The efficacy of amlodipine in myocardial ischemia. AB - Calcium antagonists are among the most potent and efficacious drugs used in the treatment of angina pectoris. Amlodipine, a new member of this family of dihydropyridines, has a unique pharmacokinetic profile with high bioavailability and an extended period of pharmacodynamic activity. In formal randomized, double blind, placebo-controlled trials with exercise tests carried out 24 hours after administration, amlodipine was significantly more effective than the placebo and comparable in efficacy with the calcium antagonist diltiazem and the beta blocking drug nadolol. In addition to extending exercise capacity in patients with angina pectoris, amlodipine significantly reduces ECG evidence of myocardial ischemia. Amlodipine has also been found to be effective in reducing the anginal attack rate in patients with vasospastic angina. From the evidence available, it is concluded that once-daily treatment with amlodipine in the dose range of 5 to 10 mg is effective in improving exercise capacity and reducing anginal attack rate in patients with chronic stable angina pectoris and also those with vasospastic angina. PMID- 2573267 TI - Ultrastructural architecture of pulmonary small-granule cell clusters in adult Syrian golden hamster. AB - Throughout the epithelial lining of the respiratory system is a class of cells with characteristics similar to Amine Precursor Uptake and Decarboxylation (APUD) polypeptide hormone-producing cells. In the intrapulmonary airways, these small granule cells (SGCs) occur either singly or in organized clusters. Although no specific peptide has yet been identified, subclasses have been postulated based on granule geometry or light microscopic staining. The present study characterizes the architectonic and cellular organization of clustered SGCs in the adult Syrian golden hamster. Two morphologically distinct cells can be defined in such clusters, "light" and "dark." Thid distinction was based primarily on differences in the electron density of the cytoplasmic matrix rather than on the remarkable variations in cellular organelles or dense-core secretory vesicles. Both cell types were normal as judged by uniform spherical nuclei, chromatin organization, and distribution of cellular organelles. The "dark" cells, however, presented the profile of a cell actively involved in synthesis with a markedly dilated perinuclear cisterna and endoplasmic reticulum. Additionally, the "dark" cells contained membrane-delimited structures containing concentric membranous whorls, clear vacuoles, and lipofuscin granules. Occasionally, cells were observed to contain features of both cell types, suggesting that they may represent a continuum of common cell lineage. Accordingly, in the absence of additional morphologic or biochemical data, the "light" and "dark" cells most probably correspond to different stages of functional activity or age-related changes of a single type of cell. Unmyelinated nerve endings were occasionally interposed between cells, but synaptic specializations were not observed. Beneath the clusters, nerve fibers were also present, but they were never observed to penetrate the basal lamina or contact any of the SGCs. Of equal occurrence were elements of the vascular system and smooth muscle, suggesting that some SGCs in the adult hamster may function in a paracrine or endocrine manner. Such knowledge is essential to any study attempting to delineate the functional role or roles of these enigmatic organoids. PMID- 2573269 TI - Somatostatinoma of the rectum. AB - We report a case of a 29-yr-old man with a rectal carcinoid that metastasized to the liver, secreting somatostatin. The patient first presented with an abdominal mass and mild diabetes, and, subsequently, rectal tumor and hypersomatostatinemia. Microscopic examination of the rectal tumor showed characteristic features of carcinoid. Multiple liver metastases were recognized. The plasma level of somatostatin was remarkably high (2,839 pg/ml). Plasma gut glucagon and pancreatic polypeptide (PP) increased with progress of the disease. He died of hepatic failure 20 months after his first admission. Immunoperoxidase staining showed that rectal tumor cells contained somatostatin-like immunoreactivity. Electron microscopy of tumor tissue showed neurosecretory (D cell) granules. Somatostatin content of the rectal tumor was very high (5,629 pg/mg). PMID- 2573268 TI - Effects of moderate alcohol intake in fixed or variable amounts on concentration of serum lipids and liver enzymes in healthy young men. AB - To assess the effects of moderate alcohol consumption on fasting serum triglyceride (TG), total cholesterol (TC), low-density-lipoprotein cholesterol (LDL-C), high-density-lipoprotein cholesterol (HDL-C), glutamate-oxaloacetate transaminase (GOT), and gamma-glutamyl transferase (GGT) concentrations, groups of normolipidemic, nonsmoking, nonathletes who were moderate drinkers aged 21-35 y and within 10% of ideal body weight consumed 40 g ETOH/d as beer (fixed drinkers) or maintained usual drinking habits (variable drinkers) for 6 wk, then abstained from all alcohol for 3 wk. A similar group of nondrinkers served as the control group. HDL-C concentrations increased significantly during alcohol consumption and decreased during abstention to initial values in both the variable and fixed drinkers. No significant difference was found between the two drinking groups. LDL-C and TC concentrations in variable drinkers were modestly lower than those in nondrinkers but not in fixed drinkers. No significant differences were found in TG, GOT, and GGT concentrations between the groups or during alcohol consumption or abstention. This study demonstrates that consumption of alcohol in fixed or variable amounts is associated with an increase in HDL-C. This increase is not due to an induction of GGT and GOT as speculated. PMID- 2573270 TI - Two-year evaluation of clinical and laboratory variables of immune function in 117 hemophiliacs seropositive or seronegative for HIV-1. AB - Fifty-nine HIV-1 antibody positive and 58 antibody negative hemophiliacs were evaluated over a 2 year study period to gain insight into the natural history and prognosis of HIV-1 disease in members of this risk group. Mean CD4 (Leu 3+) cell counts calculated at 6 month intervals decreased gradually in seropositive patients (from 403 to 311/microliters) whereas CD8 (Leu 2+) counts remained stable but above the normal range. CD4 cell counts correlated closely with advancing CDC clinical stage; CD8 numbers showed no such association, but were markedly lower in the six patients with overt AIDS. Serum P24 antigenemia was associated with low CD4 cell counts and with advanced clinical stage (58% of antigenemic and 14% of non-antigenemic seropositive patients were in stage IV). In addition to CD4 cell counts, significant reductions in Leu 11+ natural killer cell (NK) subsets and in Leu 3 + 8 - cells occurred in seropositive patients over the study period; Leu 2 + DR + cells increased significantly. When expressed as a percentage of lymphocytes, the reduction in Leu 19 + NK cells was also significant, as were the increases in Leu 4 + DR + cells and Leu 12 + 8 + B cells. In summary, declining CD4 cell numbers and percentages are valuable markers of progressive HIV-1 disease in hemophiliacs, but may not always accurately reflect the degree of disease activity. Progressive changes in additional variables such as serum P24 antigen, and numbers and percentages of NK cell subsets and (as AIDS supervenes) CD8 cell numbers, may allow more precise monitoring of HIV-1 disease. This will, in turn, facilitate the design of optimal individualized strategies for therapeutic intervention. PMID- 2573271 TI - Therapy with monoclonal antibodies to CD4: potential not appreciated? AB - The assumption that CD8+-cytotoxic cells effect graft rejection has diverted many in clinical and experimental transplantation into ignoring the tremendous potential of anti-CD4 monoclonal antibodies as immunosuppressive therapy. Experimental studies over the past several years has shown that anti-CD4 monoclonal antibodies have effects on prolonging graft survival equal to or greater than those of cyclosporine. This therapy has the unique capacity to induce tolerance to the monoclonal antibody itself, thus preventing the production both of antiidiotypic and antimouse immunoglobulin antibodies. Further, many animals develop tolerance to other antigens, including grafted tissue, raising the potential of eliminating the need for long-term immunosuppression. Synergy between anti-CD4 monoclonal antibodies and other immunosuppressives, including anti-CD8 antibodies and cyclosporine, has also been demonstrated. Continued investigation to determine the best monoclonal antibody to use in humans with respect to its epitope, immunoglobulin subclass, and capacity to deplete CD4+ cells is required to maximize the potential of this therapy before clinical trial. PMID- 2573272 TI - Molecular genetics of phenylketonuria and its implications. PMID- 2573273 TI - Genetic linkage analysis of hereditary arthro-ophthalmopathy (Stickler syndrome) and the type II procollagen gene. AB - Hereditary arthro-ophthalmopathy (AO), or Stickler syndrome, is a dominantly inherited disorder characterized by vitreo-retinal degeneration and frequently accompanied by epiphyseal dysplasia and premature degenerative joint disease. Three large families with AO were analyzed for clinical manifestations of the disease and for coinheritance of the genetic defect with RFLPs in the type II procollagen gene (COL2A1). Genetic linkage between AO and COL2A1 was demonstrated in the largest family, with a maximum LOD score of 3.52 at a recombination distance of zero. Data from a second family also supported linkage of AO and COL2A1, with a LOD score of 1.20 at a recombination distance of zero. These results are consistent with the conclusion that mutations in the COL2A1 gene are responsible for AO in these two families. In a third AO family, however, recombination between AO and COL2A1 occurred in at least one meiosis, and the data were inconclusive with respect to linkage. PMID- 2573274 TI - Allele-specific deletion in exon I of the HRAS1 gene. AB - We have detected a 6-bp deletion in the untranslated first exon of a unique HRAS1 gene cloned from lymphocyte DNA of a familial melanoma patient. The deletion is without apparent functional consequence. Using an RNase protection assay, we have demonstrated the deletion in leukocyte DNAs of individuals unrelated to the patient. In these cases, the deletion marker is specifically associated with one class of common HRAS1 allele, thereby establishing the origin of the unique allele. We discuss the means by which DNA sequence heterogeneity at other loci may be rapidly analyzed. PMID- 2573276 TI - Refined physical and genetic mapping of the NF1 region on chromosome 17. AB - A total of 15 polymorphic markers were used to construct a genetic map that encompasses the NF1 locus on chromosome 17. The markers were a subset of a large collection of chromosome 17-specific probes and were selected for marker typing in NF1 families after physical localization to the pericentric region of the chromosome. Multilocus data for a total of 17 informative NF1 families and 39 other families were included in genetic analyses. No recombination was observed between NF1 and four markers, one or more of which was informative in 86% of parents. More-refined physical mapping studies demonstrated that all four of the markers are proximal to the chromosome 17 translocation breakpoints from two NF1 patients bearing balanced translocations. The region flanking the disease locus spans a distance of 1 centimorgan (cM) in males and 9 cM in females. Close flanking markers were informative in 76% of meioses. Sex differences in recombination rates in the pericentric region were highly significant statistically. PMID- 2573275 TI - An Xp22 microdeletion associated with ocular albinism and ichthyosis: approximation of breakpoints and estimation of deletion size by using cloned DNA probes and flow cytometry. AB - Ocular albinism of the Nettleship-Falls type (OA1) and X-linked ichthyosis (XI) due to steroid sulfatase (STS) deficiency are cosegregating in three cytogenetically normal half-brothers. The mother has patchy fundal hypopigmentation consistent with random X inactivation in an OA1 carrier. Additional phenotypic abnormalities that have been observed in other STS "deletion syndromes" are not present in this family. STS is entirely deleted on Southern blot in the affected males, but the loci MIC2X, DXS31, DXS143, DXS85, DXS43, DXS9, and DXS41 are not deleted. At least part of DXS278 is retained. Flow cytometric analysis of cultured lymphoblasts from one of the XI/OA1 males and his mother detected a deletion of about 3.5 million bp or about 2% of the X chromosome. Southern blot and RFLP analysis in the XI/OA1 family support the order tel-[STS-OA1-DXS278]-DXS9-DXS41-cen. An unrelated patient with the karyotype 46,X,t(X;Y) (p22;q11) retains the DXS143 locus on the derivative X chromosome but loses DXS278, suggesting that DXS278 is the more distal locus and is close to an XI/OA1 deletion boundary. If a contiguous gene deletion is responsible for the observed XI/OA1 phenotype, it localizes OA1 to the Xp22.3 region. PMID- 2573277 TI - The gene for the human mast cell high-affinity IgE receptor alpha chain: chromosomal localization to Iq21-q23 and RFLP analysis. AB - We have used a cDNA probe for the human IgE receptor alpha chain to determine the chromosomal location for the human gene. A combination of Southern blot analysis of panels of somatic-cell hybrid DNAs and chromosomal in situ hybridization has localized the gene to the long arm of chromosome 1 at q21-q23. With this cDNA probe, we have also identified an RsaI RFLP which is inherited in a Mendelian fashion. This polymorphic marker on chromosome 1 should be valuable for studies directed at the identification both of diseases involving the IgE receptor alpha chain and of genetic factors affecting the allergic response. PMID- 2573278 TI - The human dopamine D2 receptor gene is located on chromosome 11 at q22-q23 and identifies a TaqI RFLP. AB - Human dopaminergic neurons are involved in the control of hormone secretion, voluntary movement, and emotional behavior. Mediating these effects are the dopamine D1 and D2 receptors. These macromolecules belong to a large family of related sequences known as the G protein-coupled receptors. The D2 receptors have been of special interest because they bind, with high affinity and specificity, many of the commonly prescribed antipsychotic drugs. We previously isolated a full-length cDNA clone of the rat D2 receptor. When a chromosome mapping panel was probed with the rat D2 receptor cDNA a 15-kb EcoRI restriction fragment was identified and localized to human chromosome 11. The rat cDNA was also used to clone a human genomic fragment, lambda hD2G1, which contains the last coding exon of the D2 receptor gene (DRD2) and 16.5 kb of 3' flanking sequence. Hybridization of lambda hD2G1 to a chromosome 11 regional mapping panel localized DRD2 to 11q. In situ hybridization of lambda hD2G1 to metaphase chromosomes refined this assignment to the q22-q23 junction of chromosome 11. A search for RFLPs associated with D2DR identified a frequent two-allele TaqI RFLP. PMID- 2573279 TI - The human chromogranin A gene: chromosome assignment and RFLP analysis. AB - Chromogranin A/secretory protein I (CgA) is a glycoprotein that is stored and released along with peptide hormones and neurotransmitters from several tissues, although its exact function is not known. A cDNA (gene symbol CHGA) clone was used as a probe in Southern blot analyses of human-rodent somatic cell hybrid DNAs. Discordancy analysis allowed confirmation of the assignment of the gene to chromosome 14. These results were extended using in situ chromosome hybridization, and a signal was found at 14q32. BglII digestion of genomic DNA from 28 unrelated Caucasian individuals probed with CHGA detected a two-allele RFLP with allelic frequencies of .34 and .66. PMID- 2573281 TI - Subdural empyema after endoscopic sclerotherapy. PMID- 2573280 TI - Prospective study of the standard meal provocative test in Zollinger-Ellison syndrome. AB - PURPOSE: The purpose of this work was to evaluate the proposed usefulness of a standard meal-stimulated gastrin provocative test in: (1) distinguishing Zollinger-Ellison syndrome (ZES) from antral syndromes; (2) localizing duodenal gastrinomas; or (3) suggesting that patients with multiple endocrine neoplasia type I (MEN-I) may have an increased incidence of antral syndromes. PATIENTS AND METHODS: Seventy-four consecutive patients with ZES referred to the National Institutes of Health were studied prospectively. The extent and location of gastrinomas, acid secretory studies, and the presence or absence of MEN-I were determined and correlated with the results of the gastrin response to standard meal provocative testing. RESULTS: For patients with fasting serum gastrin levels less than 1,000 pg/mL (n = 43), only 44% had a less than 50% increase over the pre-meal value, which is reported to be the typical response in ZES, and 40% had a 50% to 99% increase. Furthermore 16% had a 100% or greater increase, 9% a 150% or greater increase, and 5% a 200% or greater increase, which overlaps with values reported to be characteristic of 98%, 92%, and 46% of patients with antral syndromes. Results did not differ for patients with or without MEN-I, depend on the extent of the gastrinoma (duodenal versus pancreatic gastrinomas), the presence of previous gastric surgery or type of gastric surgery, or for patients with fasting serum gastrin concentrations greater than or equal to 1,000 pg/mL or less than 1,000 pg/mL. studies of four patients before and after resection of the gastrinoma, who prior to surgery had a greater than 100% increase in gastrin secretion after the meal, demonstrated that all patients had a less than 100% increase postoperatively even though no gastric resection was done. CONCLUSIONS: Approximately half of the patients with ZES have a greater than 50% increase in serum gastrin concentration following a standard test meal and one fifth have a 100% or greater increase. Therefore, they cannot be distinguished on this basis from patients with antral syndromes. The increased serum gastrin level after the meal in these patients with ZES appears to be due to the gastrinoma. Furthermore, the current study provides no evidence for the proposals that antral syndromes are more common in patients with MEN-I, that gastric surgery affects the meal response in patients with gastrinomas, or that the meal test is useful in localizing duodenal gastrinomas. PMID- 2573282 TI - Takayasu arteritis in a middle-aged Caucasian woman: clinical course correlated with duplex ultrasound and angiography. AB - Takayasu arteritis, an occlusive arteritis usually involving the aorta and its major branches, occurs most commonly in young oriental women. The authors report the case of a 54-year-old caucasian woman with Takayasu arteritis who initially presented with the inflammatory manifestations of fever and markedly accelerated erythrocyte sedimentation rate. Over 2 months she developed evidence of vascular insufficiency with arm claudication, diminished radial pulses, and decreased blood pressure in the arms, suggestive of an occlusive arteritis. Arteriography revealed the typical findings of Takayasu arteritis, which involved the brachial and axillary arteries but spared the aorta and its major branches. Changes in the patient's clinical course were correlated with sedimentation rate, angiography, and Duplex sonography. PMID- 2573283 TI - The spindle-shaped cells in cutaneous Kaposi's sarcoma. Histologic simulators include factor XIIIa dermal dendrocytes. AB - Kaposi's sarcoma is a neoplasm that develops as multifocal lesions, often involving the skin, characterized by a complex histologic picture including numerous vascular spaces, perivascular and interstitial spindle-shaped cells, and extravasated erythrocytes, lymphocytes, and plasma cells. Using an antibody against factor XIIIa, which identifies dermal dendrocytes, numerous factor XIIIa positive dermal dendrocytes were detected among the spindle-shaped cells in 12 acquired immune deficiency syndrome (AIDS)-associated, and five non-AIDS associated Kaposi's sarcoma lesions. The factor XIIIa-positive dermal dendrocytes were also increased in histologic simulators of Kaposi's sarcoma such as dermatofibroma, angiomatoid malignant fibrous histiocytoma, granuloma annulare, and early wound healing, but were absent in keloids. The increased number of dermal dendrocytes, which are often in an angiocentric configuration and which also express CD4, lymphocyte function associated antigen-1 (LFA-1), and Leu M3 in Kaposi's sarcoma, may be important to the angioproliferative response. The results suggested that the spindle-shaped cells that are present in a variety of cutaneous lesions are dermal dendrocytes and belong to the reticuloendothelial system, unlike other mesenchymal cell types such as the endothelial cell. Apparently a diverse array of stimuli, including human immunodeficiency virus type-1 (HIV-1) infection and trauma, can stimulate the accumulation of factor XIIIa expressing dermal dendrocytes in the skin. These cells can then participate in different stages of a variety of cutaneous alterations including Kaposi's sarcoma, dermatofibroma, granuloma annulare, and early wound healing. Thus, the factor XIIIa-positive dermal dendrocyte is a common cellular denominator among diverse clinical entities that share some histologic features. PMID- 2573284 TI - Coxsackievirus-induced disease. CD4+ cells initiate both myocarditis and pancreatitis in DBA/2 mice. AB - DBA/2 male mice inoculated intraperitoneally with 1.8 X 10(5) plaque-forming units (PFU) coxsackievirus B-3 (CVB3) showed extensive inflammatory cell infiltration of the myocardium and acinar tissue of the pancreas in 7 days. Selective depletion of T lymphocyte subpopulations indicated that CD4 cells were either completely or partially responsible for cell damage in both organs. Other organs such as the liver were infected and contained virus titers equivalent to those seen in the heart and pancreas but showed no apparent tissue injury. The role of the CD4 cell was confirmed by positive selection of either T cell subpopulation from CVB3-immune lymphocytes in vitro and adoptive transfer of these cells into T cell-deficient (thymectomized, irradiated, bone marrow reconstituted, TXBM) DBA/2 recipients. Lymphocytes from CVB3-infected donor mice were adsorbed to myocyte, skin fibroblast, or liver vascular endothelial cell (VEC) monolayers. The adherent population was retrieved and adoptively transferred into uninfected syngeneic recipients. When killed 7 days later, the animals receiving unfractionated immune lymphocytes or cells eluted from heart monolayers developed both myocarditis and pancreatitis. Anti-Thy 1.2 and C' treatment of the unfractionated cells completely abrogated transfer of disease. Cells eluted from either fibroblast or liver VEC monolayers showed no pathogenicity. Adsorption of immune cells to heart monolayers in the presence of anti-IAd (class II major histocompatibility complex antigen, MHC) inhibited attachment of the pathogenic T cell, whereas anti KdDd (a class I MHC antigen) had no effect. PMID- 2573285 TI - Neurochemical mechanisms of motion sickness. AB - Three kinds of neurotransmitters (histamine, acetylcholine, and catecholamine) are thought to be important in the neural processes of motion sickness because antihistaminics, scopolamine, and amphetamine are effective in preventing motion sickness. In this study, we examined the neurochemical and neuropharmacologic features of motion sickness in rats. Based on our results, we propose the following hypotheses for the neurochemical mechanisms of motion sickness: (1) the histaminergic neuron system is involved in the signs and symptoms of motion sickness, including vomiting; (2) the acetylcholinergic neuron system is involved in the processes of habituation to motion sickness, including neural store mechanisms; and (3) the catecholaminergic neuron system in the brain stem is not related to the development of motion sickness. PMID- 2573286 TI - Terfenadine once daily in chronic urticaria. A multi-centre double-blind comparison of terfenadine once daily versus twice daily. AB - The objective of these three double-blind multi-centre studies was to determine whether terfenadine 120 mg once daily has similar efficacy and tolerability as the standard dosage of 60 mg twice daily in the treatment of chronic urticaria. A total of 252 patients were randomly allocated to two parallel groups and treated for 2 weeks with either regimen. Evaluation of efficacy was based on rating scales for investigator and patient, i.e. itch, number of wheals, wheal size and an overall rating of efficacy. A similar improvement was seen in all variables, and there were no statistically significant differences between treatment groups. The power of the studies combined is greater than 80%, i.e. sufficient to state convincingly that treatment effects are not different. Both treatments were also equally well tolerated. PMID- 2573287 TI - Hypersensitivity pneumonitis by sulphasalazine. PMID- 2573288 TI - Antihistamines in atopic dermatitis. PMID- 2573289 TI - Ammonia assimilating enzymes from cyanobacteria: in situ and in vitro assay using high-performance liquid chromatography. AB - Assay systems for ammonia assimilating enzymes in cyanobacteria are reported. Glutamine synthetase, glutamate synthase, and glutamate dehydrogenase can be easily assayed in situ, after the cells are made permeable to the reagents, or in vitro. The method is based upon the quantitation of glutamine or glutamate after the separation, when needed, of their o-phthaldialdehyde derivatives by reverse phase high-performance liquid chromatography on a C18 column. The isocratic elution and the fluorometric detection of the amino acid derivatives make the method fast, simple, sensitive, and free of the assay artifacts which can be produced in coupled assays or when spectrophotometric measurements are carried out in the turbid samples employed for in situ assays. PMID- 2573291 TI - A mass spectrometric technique for detecting and identifying by-products in the synthesis of peptides. AB - The utility of a new mass spectrometric technique for detecting and identifying peptide by-products produced in the synthesis of peptides is demonstrated. The technique involves three sequential steps: (1) practically nondestructive 252Cf plasma desorption mass spectrometric analysis of monolayer amounts of the peptide(s) of interest bound to a thin layer of nitrocellulose; (2) enzyme catalyzed microscale chemical reaction of the surface-bound peptide(s) to produce structurally informative hydrolysis products; (3) plasma desorption mass spectrometric analysis of these hydrolysis products. The first step determines the presence and the molecular weights of unwanted by-products resulting from errors or incomplete reactions during synthesis. The subsequent two steps provide information on the precise location in the peptides where errors have occurred. In the present paper, the technique is applied to an investigation of unwanted peptide by-products associated with the use of tryptophan during stepwise solid phase peptide synthesis. Synthetic preparations of melittin and [Bpa-8]dynorphin A (1-17) were each found to contain a major impurity with molecular weight 28 Da higher than that of the desired product. The impurity in the melittin preparation, in which the final deprotection step involved the high-low HF procedure, was shown to result from incomplete removal of the formyl group from Trp-19. On the other hand, the impurity in the [Bpa-8]dynorphin A (1-17) preparation, where the removal of the formyl group from Trp-14 was carried out using piperidine, was shown to result from migration of the formyl group to Lys 11 or Lys-13.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573290 TI - A glutamate dehydrogenase-based method for the assay of L-glutamic acid: formation of pyridine nucleotide fluorescent derivatives. AB - A method for the quantitation of L-glutamic acid in the picomole range was developed by finding conditions which allowed the production of NADH by the action of the L-glutamate dehydrogenase (EC 1.4.1.3) and its subsequent transformation to a highly fluorescent derivative. The method measures linearly glutamate from 250 pmol to 5 nmol. For its simplicity and low cost it is ideally suited to the assay of a large number of samples within a single working day. Its application to the determination of regional glutamate levels in the rat brain, as well as to the measurement of ornithine aminotransferase (EC 2.6.1.13) activity from several tissues is described. The results are similar to those obtained by different methodologies in several laboratories, but the present method offers additional advantages. PMID- 2573292 TI - Quantitation of tyrosine hydroxylase, protein levels: spot immunolabeling with an affinity-purified antibody. AB - Tyrosine hydroxylase was purified from bovine adrenal chromaffin cells and rat pheochromocytoma using a rapid (less than 2 days) procedure performed at room temperature. Rabbits were immunized with purified enzyme that was denatured with sodium dodecylsulfate, and antibodies to tyrosine hydroxylase were affinity purified from immune sera. A Western blot procedure using the affinity-purified antibodies and 125I-protein A demonstrated a selective labeling of a single Mr approximately 62,000 band in samples from a number of different tissues. The relative lack of background 125I-protein A binding permitted the development of a quantitative spot immunolabeling procedure for tyrosine hydroxylase protein. The sensitivity of the assay is 1-2 ng of enzyme. Essentially identical standard curves were obtained with tyrosine hydroxylase purified from rat pheochromocytoma, rat corpus striatum, and bovine adrenal medulla. An extract of PC 12 cells (clonal rat pheochromocytoma cells) was calibrated against purified rat pheochromocytoma tyrosine hydroxylase and used as an external standard against which levels of tyrosine hydroxylase in PC12 cells and other tissue were quantified. With this procedure, qualitative assessment of tyrosine hydroxylase protein levels can be obtained in a few hours and quantitative assessment can be obtained in less than a day. PMID- 2573293 TI - The effect of non-union of testis and epididymis and of cryptorchidism on the development of epididymis and ductus deferens in the rat. AB - 16-days old rats were operated with either uni- or bilateral ligation of ductuli efferents and separation of testis and epididymis to the level of the inferior epididymal artery (non-union operation), induction of cryptorchidism or bilateral sham operation. The epididymides were weighed and the epididymides and deferent ducts were examined with light- and electron-microscopy at days 30, 37, 44 and 58. Bilateral non-union operated epididymides and cryptepididymides had a significantly lower weight increase than controls, but the histology and diameter of epididymal tubules were unchanged. This indicates a true growth retardation and reduced length of epididymal tubules of non-union operated and cryptepididymides. For bilateral operations a positive correlation was found between the weight of epididymis and plasma levels of total testosterone as reported earlier. Unilaterally operated epididymides had a weight development significantly below contralateral controls, despite normal plasma levels of testosterone. It is concluded that the reduced-weight of unilaterally operated epididymides is the result of diminished local androgen stimulation from the ipsilateral testis. Non-union of testis and epididymis may have pathogenetic significance in maldescent of testis by a retarded growth of the ductal system. PMID- 2573294 TI - Maternally administered esmolol produces fetal beta-adrenergic blockade and hypoxemia in sheep. AB - Although esmolol may be a useful therapeutic agent in obstetrics and obstetric anesthesia, concerns about fetal safety have limited its use. To assess acute fetal hemodynamic effects of maternally administered esmolol, saline or esmolol (4-200 micrograms.kg-1.min-1 in a stepped manner) was infused into maternal venous catheters in nine chronically prepared pregnant ewes, and the degree of beta-adrenergic blockade was assessed by isoproterenol challenge. In control experiments saline infusion and repeated isoproterenol challenges did not alter measured parameters, although maternally administered isoproterenol (0.1 micrograms) transiently decreased uterine blood flow by 20 +/- 5% (mean +/- SEM; P less than 0.05). Esmolol produced a dose-dependent decrease in maternal blood pressure and fetal heart rate (maternal blood pressure decreased by 22 +/- 8% and fetal heart rate decreased by 27 +/- 7% following esmolol, 200 micrograms.kg 1.min-1; P less than 0.05). Fetal arterial PO2 decreased from 18.2 +/- 1.2 mmHg before to 14.1 +/- 1.5 mmHg following esmolol, 200 micrograms.kg-1.min-1 (P less than 0.05). Maternally administered esmolol produced similar dose-dependent beta adrenergic blockade in both ewe and fetus, with complete blockade following the 80 and 200 micrograms.kg-1.min-1 doses. Thirty minutes following cessation of esmolol infusion, fetal resting heart rate and maternal and fetal isoproterenol stimulated heart rate remained below control values. These results suggest that maternally administered esmolol may produce adverse fetal effects, limiting its usefulness in the obstetric setting. PMID- 2573296 TI - The European experience with antihistamines in asthma. AB - Until the appearance of nonsedating antihistamines such as terfenadine and astemizole, clinical studies of H1 antagonists in the treatment of asthma yielded disappointing results. The author reviews recent European studies that demonstrate terfenadine's effectiveness in reducing symptoms and improving bronchodilation in patients with mild atopic asthma. PMID- 2573295 TI - S-Mg does not change inversely to S-FFA during acute stress situations. AB - The apparently divergent changes in serum magnesium (S-Mg) and serum free fatty acids (S-FFA) in stress situations associated with elevated levels of circulating adrenaline were studied experimentally in 12 healthy volunteers, who were each given three adrenaline infusions (0.05 micrograms/kg bw/min over one hundred twenty minutes). Before the adrenaline infusions the volunteers were treated for three days with either a non-selective beta-blocking agent (propranolol) or a beta-1-selective agent (atenolol) or with placebo. Six of the volunteers underwent a fourth adrenaline infusion after pretreatment with a beta-2-selective beta-blocking agent (ICI 118551). S-Mg and S-FFA were determined every fifteen minutes. After pretreatment with placebo, adrenaline infusion caused an increase in S-FFA from 0.22 +/- 0.20 mmol/L (mean +/- SD) to max 0.59 +/- 0.39 mmol/L after thirty minutes of adrenaline infusion (p less than 0.001). At one hundred twenty minutes S-FFA had decreased to 0.35 +/- 0.26 mmol/L. Thirty minutes after cessation of the adrenaline infusion, S-FFA had returned to the same level as before the infusion. S-Mg before the adrenaline infusion was 0.83 +/- 0.05 mmol/L, rose to 0.85 +/- 0.05 at fifteen minutes, and then decreased to 0.78 +/- 0.05 at one hundred thirty-five minutes (p less than 0.05). Pretreatment with atenolol did not change this pattern, although the changes in S-FFA and S-Mg were of a smaller magnitude but still statistically significant. PMID- 2573297 TI - Allergenicity of Chironomidae in asthmatic patients. AB - In order to clarify the relation between asthma and Chironomidae, we examined the cross-reactivity between Chironomidae and other common allergens. We noted significant correlations between positive skin tests with Chironomidae and with other allergens. The radioallergosorbent inhibition test, however, suggested that there may be no cross-reactivity or, if any, only very low cross-reactivity between midge allergens and mite, house dust (HD), silk, shrimp, or mosquito allergens. PMID- 2573298 TI - Procaterol metered-dose inhaler: a multiclinic study evaluating the efficacy and safety in patients with asthma. AB - Procaterol hydrochloride aerosol, a potent beta 2-adrenergic bronchodilator, was evaluated in a double-blind, placebo-controlled study for efficacy and safety in 210 patients with documented mild to moderate reversible airway obstruction. Patients were randomized to receive procaterol in two inhalations (high dose) or one inhalation (low dose), 0.01 mg/inhalation, three times daily, or placebo. Pulmonary function tests were recorded at five and 30 minutes and hourly for eight hours after the first dose and following 1 and 2 weeks of treatment. Both doses of procaterol produced significantly greater improvement in PFTs at one hour and for up to seven hours after dosing compared with placebo (p less than 0.05). Mean percent increases in FEV1 were 35% in the high-dose group and 29% in the low-dose group at week 2. The high-dose group showed no loss of duration of bronchodilation with continued dosing. Improvement in PFTs and peak flow rates was significantly greater in the high-dose than in the low-dose group (p less than 0.05). Tremor was the most frequent side effect. Procaterol had no effect on electrocardiograms, heart rate, blood pressure, or clinical laboratory tests. The high dose of procaterol aerosol was shown to be an effective and well-tolerated bronchodilator with a rapid onset and long duration of action. PMID- 2573299 TI - The changing role of antihistamines in asthma. Symposium held at the 45th annual congress of the American College of Allergy and Immunology. November 16, 1988. Proceedings. PMID- 2573300 TI - Terfenadine blockade of exercise-induced bronchospasm. AB - Terfenadine, in doses of 60, 120, and 180 mg, provided significant bronchodilation for up to four hours after administration. In addition, when the same doses were given four hours prior to exercise, the drug demonstrated significant blockade of exercise-induced bronchospasm at two and five minutes postexercise. These effects on exercise-induced bronchospasm followed a dose response curve; 180 mg had the most significant effect. All dosing levels, however, produced both bronchodilation and a significant degree of blockade of exercise-induced bronchospasm. No patients were bothered by adverse effects during this single-dose study. Eight of the 11 subjects who completed the study also noted some improvement in their nasal congestion during the testing period when they were on the active drug. PMID- 2573301 TI - In vivo and in vitro effects of antihistamines on mast cell mediator release: a potentially important property in the treatment of allergic disease. AB - A nasal antigen challenge model of allergic individuals was used to evaluate whether antihistamines could inhibit human mast cell and basophil mediator release in vivo. In placebo-controlled trials, topically applied azatadine base, a tricyclic antihistamine with in vitro antirelease action, effectively reduced symptoms and mediator levels in nasal lavage fluids after antigen challenge, suggesting mast cell inhibition. Both terfenadine and cetirizine, systemically administered antihistamines, were clinically effective in reducing sneezing and changes in vascular permeability. Only terfenadine significantly reduced histamine in antigen-induced nasal secretions. However, cetirizine did reduce the level of leukotriene C4 in these fluids. These results indicate that some antihistamines may be capable of suppressing mediator release from nasal mast cells. The significance of this property in those compounds' overall clinical effect is unclear because of their other concomitant activities. PMID- 2573302 TI - [Digestive stress hemorrhage. Physiopathology and prevention]. AB - Lesions of the gastroduodenal mucosa are seen very early on in virtually 100% of patients suffering from organ failure. Bleeding, even if it is only occult, defines acute stress-induced gastrointestinal tract bleeding (SGIB). The rates of SGIB vary according to the inclusion criteria: 13 to 100% microscopic SGIB, 2.3 to 9.5% haemorrhage with blood transfusion and/or shock. Gastrointestinal bleeding does not really influence the death rate of patients with SGIB (0 to 5% increase). Damage to the gastric mucosa may be due to an intraluminal aggression, and/or decreased mucosal and mural defence mechanisms. H+ ions and bile salts are mostly responsible for the former. Physiological quantities of H+ ions may be sufficient, as their abnormal diffusion into the gastric mucosa will reduce the mucosal pH (pHm), which is itself sensitive to microcirculatory modifications and systemic acidosis. There is a good correlation between bleeding and pHm. Bile salts are involved because of the usual increase in frequency and volume of gastric biliary reflux due to stress. Surfactant, mucosal alkaline layer and the microcirculation are all involved in gastric protection. The PGE2 synthetized by the gastric mucosa have a favourable influence on these 3 mechanisms. Changes in microcirculation and hypoxia are the predominant factors involved in stress induced mucosal damage. The prevention of SGIB relies on the treatment of risk factors, a reduction of intraluminal aggression, and the support and/or stimulation of gastric defence mechanisms. Antacids and anti-H2 drugs aim to neutralize most of the H+ ions, being more efficient than placebo in increasing gastric pH greater than 4, although anti-H2 agents are responsible of a greater number of failures. The non-homogenous character of the patient groups studied and the diagnostic methods, as well as the increasing lack of placebo groups in the published studies make the interpretation of the results rather risky. Antacids and anti-H2 drugs are more efficient than placebo, and equally efficient, in preventing overt SGIB. Efficiency is increased by giving anti-H2 drugs continuously, and antacids hourly. Other agents are thought to protect mucosal cells, probably increasing mucosal defences. Amongst them are the prostaglandins, the most interesting of which are still being investigated, and sucralfate. The latter molecule is as efficient as antacids and anti-H2 drugs, and does not alter gastric pH, so reducing the number of nosocomial pneumonias. Its reduced cost and easy administration make it, at the present time, the treatment of choice of SGIB. The few rare contraindications of sucralfate will justify the infusion of anti-H2 drugs in those patients at risk. PMID- 2573304 TI - Multiple endocrine neoplasia type 2a associated with cutaneous lichen amyloidosis. AB - PURPOSE: To describe and characterize the association of hereditary cutaneous lichen amyloidosis with multiple endocrine neoplasia type 2a (MEN 2a). DESIGN: Survey of a family for two diseases. SETTING: Evaluation of patients at a clinical research center. PATIENTS: Nineteen family members with MEN 2a. MEASUREMENTS AND MAIN RESULTS: In this family cutaneous lichen amyloidosis presented as multiple infiltrated papules overlying a well-demarcated plaque in the scapular area of the back (right or left). Immunohistochemical studies showed amyloid that stained for keratin but not calcitonin. Three family members had the characteristic skin lesion and also carried the gene for MEN 2a; two additional members carried the gene for MEN 2a, but did not manifest the observable skin changes associated with lichen amyloidosis. CONCLUSIONS: From the findings in this kindred and in another recently reported but unrelated family with an identical type of pruritic skin rash and MEN 2a, several conclusions can be drawn. First, the syndrome of cutaneous amyloidosis and MEN 2a is a clearly defined autosomal dominant hereditary syndrome. Second, the dermal amyloid appears to be caused by deposition of keratin-like peptides rather than by calcitonin-like peptides. Third, known families with hereditary lichen amyloidosis should be screened to determine the true frequency of this syndrome. PMID- 2573303 TI - Mechanism of reduced cardiac output during positive end-expiratory pressure in the dog. AB - The decrease in left ventricular (LV) stroke volume during positive end expiratory pressure (PEEP) has been attributed to reduced LV filling and a decreased contractile state. To assess the relative importance of each mechanism, we examined the effects of zero, 5, 10, and 15 cm H2O PEEP on LV end-diastolic volume (EDV) and end-systolic volume (ESV), and on LV contractile performance using instantaneous pressure-volume loops recorded with micromanometer-tipped and volume conductance catheters in the LV. The LV contractile state was determined using the LV end-systolic pressure-volume relations, the LV dP/dtmax-EDV relation, and the LV stroke work-EDV relation. The importance of autonomic reflexes was assessed by repeating the sequence of PEEP after beta-adrenergic blockade using metoprolol, 10 mg administered intravenously. LV EDV decreased from a baseline of 37.8 +/- 3.7 ml (+/- SEM) to 35.0 +/- 3.8, 28.7 +/- 2.9, and 25.9 +/- 2.6 ml with 5, 10, and 15 cm H2O PEEP, respectively (p less than 0.05 for each comparison), which paralleled the decline in stroke volume. In contrast, LV ESV did not change significantly with PEEP. The slope and position of the LV end-systolic pressure-volume relation and the slopes of the LV dP/dtmax-EDV relation and the LV stroke work-EDV relation (p = NS) were not altered during PEEP before or after beta-adrenergic blockade, indicating no depression of systolic contractile function. We conclude that the decreased LV stroke volume that occurs with PEEP is due to impaired LV filling (i.e., reduced LV EDV) without a concomitant depression of myocardial contractility. PMID- 2573305 TI - Pancreatitis and gold treatment of rheumatoid arthritis. PMID- 2573307 TI - Diaphragmatic dysfunction after open heart surgery: treatment with a rocking bed. AB - Of 1225 patients undergoing open heart surgery over an 18-month period, 13 had diaphragmatic dysfunction due to phrenic nerve injury; 11 of these had internal mammary artery grafting. Nine had diaphragmatic dysfunction on the same side as the internal mammary artery graft side (7 bilateral and 2 unilateral) as determined by fluoroscopy during phrenic nerve stimulation. Although topical cardiac hypothermia has been the prevailing mechanism for diaphragmatic dysfunction due to phrenic nerve injury after open-heart surgery, dissection of the internal mammary artery with electrocautery, traction, or vascular compromise to the phrenic nerve, or a combination, could be additional factors. Rocking bed ventilation was instituted to facilitate passive diaphragmatic movement and airway decannulation and was continued at home until the phrenic nerve or nerves recovered. These patients were followed up clinically and with serial measurements of vital capacity, respiratory muscle strength, phrenic nerve latency, and fluoroscopy to determine recovery rate. Phrenic nerve recovery occurred from 4 to 27 months after surgery. This recovery was heralded by the patients' ability to assume the supine position without dyspnea when use of the rocking bed was discontinued. Unilateral diaphragmatic recovery was sufficient for the restoration of symptom-free supine posture. PMID- 2573306 TI - Human T-cell lymphotropic virus type I (HTLV-I)-associated adult T-cell leukemia lymphoma in a patient infected with human immunodeficiency virus type 1 (HIV-1). AB - A patient had adult T-cell leukemia-lymphoma in the unusual setting of coinfection with human immunodeficiency virus type 1 (HIV-1) and human T-cell lymphotropic virus type I (HTLV-I). The leukemic cells were CD4 positive and showed clonal genetic rearrangement of the T-cell receptor complex. Cytogenetic analysis showed three clonal karyotypic abnormalities: trisomy 3 and two translocations [t(1;15), (X;1)]. The patient was seropositive for HIV and HTLV-I; HTLV-I and HIV-1 DNA sequences were detected in peripheral blood leukocytes by the polymerase chain reaction. The HTLV-I sequences were detected in a relatively high proportion of mononuclear cells (at least 1 in 30 cells), whereas HIV-1 sequences were detected in a smaller proportion of cells (at least 1 in 3000 cells). Clinical remission was achieved after chemotherapy. There was a decrease in the proportion of HTLV-I positive mononuclear cells (at least 1 in 1000 cells), whereas the proportion of HIV-1 positive cells was relatively unchanged (at least 1 in 1000 cells). Adult T-cell leukemia-lymphoma in the setting of HIV coinfection may become increasingly common because asymptomatic retroviral coinfections are frequent. PMID- 2573308 TI - [Cutaneous manifestations of Takayasu arteritis. Apropos of a case of Bazin's erythema induratum]. PMID- 2573309 TI - [Respiratory arrest in an asthmatic girl treated with beta-2-mimetics and theophylline. Possible role of hypokalemia in sudden death in asthmatic patients]. AB - We report a case of respiratory arrest in a ten-year-old asthmatic girl under beta-2-agonists and theophylline. The possibility that hypokalemia plays a part in the sudden deaths seen in some asthmatic patients is discussed. PMID- 2573310 TI - [Experimental equine fascioliasis: evolution of serologic, enzymatic and parasitic parameters]. AB - Three, four, and one horses were respectively infected with 100, 1,000, and 5,000 metacercariae of Fasciola hepatica. Six of them were reinfected 38 weeks later with 1,000 metacercariae each. Specific antibodies assayed by counter electrophoresis, passive hemagglutination and ELISA tests appeared three to six weeks post-infection and peaked 10 to 17 weeks post-infection. Horses infected by 1,000 metacercariae and more showed 17.6% of positive samples by counter electrophoresis, 49.2% by ELISA, and 75.6% by passive hemagglutination. Plasma glutamate dehydrogenase and gamma-glutamyltransferase levels increased significantly 3 to 5 months post-infection in the most infected animals. Eggs of Fasciola hepatica were only observed in 2 of the 8 horses, 14 and 15 weeks post infection. This last observation indicates the limits of fecal examination in the diagnosis of fascioliasis in horses. PMID- 2573311 TI - Effect of body mass and other factors on serum liver enzyme levels in men attending for well population screening. AB - Alcohol abuse is usually regarded as the most likely cause of elevated serum liver enzyme values in those attending for well population screening, but we have found increased body weight to be an important contributing factor. We have measured serum levels of alanine amino-transferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyl transferase (GGT) in approximately 21,000 men attending for routine health screening, and related these to behavioural factors such as alcohol consumption, cigarette smoking, exercise level and obesity. The levels of all three enzymes were positively correlated with levels of alcohol consumption. Decreasing levels of physical activity were associated with increases in mean ALT and GGT levels. Cigarette smoking showed only a weak effect on ALT and AST, which became non-significant after multivariate statistical analysis, but increasing consumption of cigarettes was associated with increased mean levels of GGT. In contrast, all three enzymes showed marked increases in mean levels with increasing body mass index (BMI). The effect of obesity was particularly important in the case of ALT: the prevalence of increased ALT values in obese subjects (BMI greater than or equal to 31 kg/m2) was more than eight times that in those with normal weight (BMI less than or equal to 25 kg/m2), even after allowing for the confounding effect of alcohol consumption. This study is concerned solely with male subjects, but we hope to extend the analysis to females in the near future. PMID- 2573312 TI - Unsuitability of the human erythrocyte as a model for in vivo sodium transport activation by beta 2 adrenergic agonists. AB - Administration of the beta 2 adrenergic agonists fenoterol, salbutamol and terbutaline to volunteers significantly reduced plasma potassium concentration in a double-blind crossover study. There was no consistent effect of the three active compounds on erythrocyte sodium concentration. Storage of whole blood at 4 degrees C increased erythrocyte sodium concentration by 33% after 2 h; this could explain the differences between this and another study of the effects of beta 2 adrenergic agonists on erythrocyte sodium concentration. We conclude that the human erythrocyte is unsuitable as an indirect in vivo model to demonstrate stimulation of cellular sodium potassium transport mediated by beta 2 adrenergic agonists. PMID- 2573313 TI - Normal phenotype and slight mental retardation in de novo distal 8p deletion (8pter----8p23.1:). AB - In this report we present a 9-year-old boy with mental retardation, behavioural problems and terminal deletion of the short arm of chromosome 8(8pter--- 8p23.1:). In contrast with previously reported patients with larger terminal and interstitial 8p deletions he did not present major phenotypic abnormalities. PMID- 2573314 TI - Partial duplication of the short arm of chromosome 2 (dup(2)(p13----p21) associated with mental retardation and an Aarskog-like phenotype. AB - In this report we describe a 3-year-old boy with partial trisomy of the short arm of chromosome 2 due to a de novo tandem duplication 2(dup(2)(p13----p21)). In addition to severe growth retardation and moderate psychomotor delay he presented a dysmorphic syndrome compatible with the clinical diagnostic of Aarskog syndrome. PMID- 2573315 TI - In vivo effect of a beta-adrenergic agonist on activity of calcium-dependent proteinases, their specific inhibitor, and cathepsins B and H in skeletal muscle. AB - DEAE-Sephacel and phenyl-Sepharose chromatography were compared as methods for separating and quantitatively isolating calpain I, calpain II, and calpastatin from lamb muscle extracts. DEAE-Sephacel chromatography gave greater than 90% recovery of all three proteins, while phenyl-Sepharose gave only 70, 66, and 48% of the DEAE recovery of calpain I, calpain II, and calpastatin, respectively. Additionally, DEAE-Sephacel chromatography was shown to effectively separate calpastatin and calpain I. Consequently DEAE-Sephacel appears to be superior to phenyl-Sepharose for quantitative isolation of the components of the calcium dependent proteinase system from muscle extracts. Dietary administration of beta agonist (L-644, 969; Merck Sharpe & Dohme Research Laboratories) decreases extractable calpain I activity in lamb longissimus dorsi (LD) muscle by 10-14% (P less than 0.05), increases calpain II activity by 34-42% (P less than 0.001), and increases calpastatin activity by 59-75% (P less than 0.001). Additionally, net cathepsin B activity is reduced by 30% (P less than 0.05) in the LD of beta agonist-treated lambs. Reduced activity of the calcium-dependent or catheptic proteinase systems may contribute to the increased protein accretion in muscles of beta-agonist-treated lambs. PMID- 2573316 TI - Intracellular Ca2+ and cytotoxicity. AB - Following injury or activation in some immune cell lines, elevation of intracellular Ca2+ concentration (Cai2+) is an early and major event that precedes cell death. Agents shown to elevate Cai2+ and to result subsequently in the death of some cells include human immunodeficiency virus (HIV) (in T4+ cells), 25-hydroxy cholesterol, tumor necrosis factor (TNF), cyclosporine, dexamethasone, alpha-interferon, and Ca2+ ionophores. The effects of these agents, both on Cai2+ and on cytotoxicity, are additive. This type of Ca2+ related cytotoxicity may be associated with either accelerated synthesis of triglycerides (TNF), accelerated synthesis of cholesterol ester (25-hydroxy cholesterol), or cholesterol (HIV) and terminally with declining synthesis of structural phospholipid. Agents that can lower Cai2+ (e.g., phorbol esters, diglycerides, lipoproteins [LDL], oleic acid, or serum) under appropriate conditions ameliorate the Ca2+-induced cytotoxicity. Metabolism of other divalent metals, i.e., Zn2+ and Cd2+, also become altered with cell injury, e.g., glucocorticoids elevate Cai2+, but block uptake of Zn2+. These observations support the idea that chronic elevation of Cai2+ by many chemically unrelated agents leads to cell death by creating imbalance both in cell biosynthetic mechanisms--especially in those controlling lipid metabolism--as well as creating imbalances in metabolism of other trace metals, especially Zn2+. PMID- 2573317 TI - [Persistent Mullerian duct syndrome]. AB - We report on a 14-year-old boy who was admitted to the emergency department for acute retention of urine. At age 1 year, he had been submitted to surgery for left cryptorchidism and left-sided herniation, associated with hypospadias of the glans penis. Radiologic and endoscopic work up revealed a large retrovesical cavity with septation in its upper portion communicating with the prostatic urethra. These structures were removed and pathologic examination disclosed remnants of Mullerian ducts. This uncommon disease entity whose etiopathogenesis is ascribed to a deficient activity of the Mullerian inhibiting factor (MIF) classically presents in phenotypically male subjects with unilateral cryptorchidism, contralateral herniation, and persistent Mullerian remnants. The literature is reviewed, highlighting the diagnostic and therapeutic aspects of this syndrome. PMID- 2573318 TI - [Seminoma in the undescended testis]. AB - Maldescend testes are predisposed to tumor degeneration from embrional germinal dysgenesis, the cause of cryptorchidism. The risk of carcinogenesis is thirty to fifty times greater than in normal descended testes. The histologic features of this tumor type and its treatment do not differ from those of normotopic tumors. PMID- 2573319 TI - Polyarteritis nodosa associated with idiopathic pulmonary fibrosis: report of two cases. PMID- 2573320 TI - Pharmacological responses of internal mammary artery grafts. PMID- 2573322 TI - [Open trial of betaxolol in the treatment of arterial hypertension in pregnancy, pilot study]. AB - The efficacy and the safety of betaxolol (BTX), a cardio-selective adrenoceptor blocking agent with a long half life, was tested in 22 hypertensive pregnant women. BTX doses varied from 10 to 40 mg/day. BTX given as monotherapy (n = 19) or associated to dihydralazine (n = 3) led to a satisfactory control of arterial blood pressure (ABP). In the mother, the drug was very well tolerated. (table; see text) The fetal safety, evaluated by mean of cardiotocography and echography was also satisfactory. Fetal heart rythm was significantly slowed following first intake of BTX and remained stable until delivery; there was only one case of intra-uterine growth retardation pre-existing to the pharmacological treatment. Nine deliveries were ceasarian [fetal distress (n = 1); poor control ABPI (n = 1); obstetrical reasons (n = 7)]. Seventeen neonates were full-term and six were premature (twins included). The state of neonates at birth was very satisfactory (Apgar at 5 minutes greater than 9 in 21/23). Glycemia less than 1.66 mmoles/1 in two neonates, was normal within 24 to 48 hours. In conclusion, BTX was very efficacious in controlling increased blood pressure in pregnant women and its safety in the mother, the fetus and the newborn was very good. PMID- 2573321 TI - [Takayasu arteritis: usefulness of radiologic and angiographic studies]. AB - With the purpose of learning the usefulness of radiologic and angiographic procedures for the evaluation of Takayasu's arteritis (TA), we studied 64 patients (ratio women: 8.1, average age: 23.5 years, range: 13-52 years) in which we performed arteriographic studies in the clinically affected area. All cases had chest films: 53 had thoracic aortogram, 60 abdominal aortogram, 16 pulmonary arteriography. According to the topography of the lesions we found 8% of the cases with damage exclusive to the supra-aortic trunk, 6% with isolated alteration of the intermediate thoraco-abdominal aorta, 62% with mixed pathology of the two categories above, and 21% with lesions in the pulmonary artery besides systemic arteriopathy. The results were as follows: 1) radiology of the chest: cardiomegaly (48%), irregularities in the ascending aorta (31%), calcification in the aortic wall (29%), calcified granulomas (25%) and signs of pulmonary venous hypertension (21%); 2) thoracic aortogram: irregularities in the descending aorta (56.6%), thickening of the wall of descending aorta (39.6%), dilatation of the ascending aorta (26.4%), of the descending aorta (26.4%); occlusions: of the left subclavian (24 cases), left mammary (16 cases), left carotid (8 cases) and left vertebral (8 cases); 3) abdominal aortogram: irregularities of the outline, stenosis, prominent "supplementary" arteries and aneurysms in 53%, 43.3%, 38% and 13.3 of the studies performed. The arteries most commonly affected were: renal (74.7%), both (31.6%), right (28.2%) and left (14.9%), superior mesenteric (26.6%) and hepatic (21.6%); 4) pulmonary arteriography: arterial occlusions: right superior lobar branch (37.5%), right medial (6.2%), right inferior (12.5%), without predilection by any lobe; 5) coronary arteriography: one case with occlusion of anterior descending artery and circumflex coronary artery (the other 8 cases without significant lesions). We concluded that TA affects independently the arteries of different areas, hence it is necessary to perform multiple angiographic studies for adequate evaluation of the extension of vascular damage. PMID- 2573323 TI - [Selectivity of alpha 2-adrenergic agonists for the imidazoline-guanidine and alpha 2-adrenergic receptors]. AB - Imidazolines have been proposed as highly selective drugs for alpha 2-adrenergic receptors. However, we have recently showed that the imidazoline ligand 3H-RX 781094 (idazoxan) binds to both alpha 2-receptors and imidazoline guanidinium receptive substance (IGRS) in rabbit renal proximal tubule. Binding of 3H-RX 781094 to the purified basolateral membranes (15-fold enriched in Na-KATPase activity) was rapid (t 1/2 = 5 mn.) reversible (t 1/2) = 4 mn.), saturable and of high affinity. Scatchard analysis of equilibrium binding data showed that 3H-RX 781094 labels 566 +/- 118 fmol/mg of proteins of binding sites with an apparent dissociation constant (Kd) of 1.45 +/- 0.14 nM. On the other hand, the non imidazoline ligand 3H-rauwolscine binds only to the alpha 2-adrenergic receptors with a maximal density of 155 +/- 28 fmol/mg of protein and a Kd of 11.5 +/- 1.5 nM. In order to define the relative affinity of the alpha-2-agonists, clonidine, rilmenidine and guanfacine for the two classes of receptors, we performed competition studies of the alpha 2-antagonists 3H-RX 781094 (imidazoline) and 3H rauwolscine (non imidazoline) binding to basolateral membranes from rabbit proximal tubule. The order of potency for inhibition of the two radioligand binding was rilmenidine greater than clonidine greater than guanfacine for 3H-RX 781094 and clonidine greater than guanfacine greater than rilmenidine for 3H rauwolscine. Therefore, rilmenidine displayed a higher affinity for IGRS than for alpha 2 adrenergic receptors; on the other hand, clonidine and guanfacine preferentially interact with alpha 2 receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573324 TI - [Vasodilator effect of tertatolol on isolated rat kidney]. AB - Tertatolol, a non cardioselective beta-blocker, maintains or increases renal blood flow in animals and in man. In this work, we confirmed in vitro the renal vasodilator effect of tertatolol and we investigated its mechanism. The rat kidney was isolated and perfused at constant flox in an open circuit with a Krebs Henseleit solution. A vascular tone was restablished by sequential bolus injections of serotonine (every 5 mn) at a dose increasing perfusion pressure by 40 mm Hg. Tertatolol and other drugs were perfused at increasing concentrations during successive periods of 15 mn. Any relaxation was expressed as percentage inhibition of the initial vaso-constriction induced by serotonine. Tertatolol (3 x 10(-7) to 3 x 10(-5) M) induced a concentration-dependent renal vasodilatation (CI50 = 4.6 +/- 0.4 x 10(-6) M, n = 5) with a maximum effect of nearly 100 p. 100 relaxation. (-) tertatolol and (+) tertatolol were respectively more and less active (CI50 = 1.7 +/- 0.3 x 10(-6) M and 10.6 +/- 2.5 x 10(-6) M, n = 5). Tertatolol metabolites (4-OH tertatolol and sulfoxytertatolol) were inactive, excluding a vascular effect of tertatolol related to its metabolism into vasoactive derivatives. Other beta-blocking drugs, (+/-) sotalol and (+/-) nadolol, were also inactive on the renal vasculature and (-) bunolol only induced renal vasodilatation at concentrations 40 fold higher than that of (-) tertatolol.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573325 TI - [Evaluation of cicletanine efficiency and tolerance in hypertensive patients treated with beta-blockaders]. AB - Efficacy and tolerance of cicletanine, the first derivative of furopyridines have been studied in a 3 parallel groups double blind study; 120 essential mild hypertensive patients uncontrolled by beta blocking therapy were included. After a period of one month during which a treatment by placebo was added to beta blocking therapy, the patients were randomly allocated into 3 groups: group I (placebo: 40 patients), group II (cicletanine 50 mg/d: 41 patients), group III (cicletanine 100 mg/d: 39 patients). Eight patients were withdrawn in the group I (poor efficacy, n = 5 or unexpected event, n = 3), and one in the group III (unexpected event). At the beginning of the treatment period (DO), lying blood pressure (LBP) was respectively for groups I, II and III 171 +/- 14/104 +/- 6, 174 +/- 13/104 +/- 5, 172 +/- 15/105 +/- 6 mmHg. 171 +/- 14/104 +/- 6, 174 +/- 13/104 +/- 5, 172 +/- 15/105 +/- 6 mmHg. After 3 months of treatment LBP was respectively 168 +/- 21/97 +/- 10, 151 +/- 12/85 +/- 7, 147 +/- 14/82 +/- 9 mmHg. A significant treatment effect (p less than 0.001) was observed on SBP with groups II and III only, and on DBP with the 3 groups. This lowering effect was comparable in groups II and III, and greater (p less than 0.001) in these groups than in group I. At the end of this study the percentage of patients whose BP was normalized was respectively for groups I, II and III: 5 p. 100, 51 p. 100 and 74 p. 100. Clinical and biological tolerance were good.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573326 TI - [Efficacy of the combination of low doses of beta-blockers and amiodarone in the treatment of refractory ventricular tachycardia]. AB - Thirty one patients aged 56 +/- 16 years with chronic ventricular tachycardias (VT) refractory to 4.4 +/- 1.8 antiarrhythmic drugs, used alone or in combination, were managed by low doses of beta-blocker agents combined with oral amiodarone, either after loading (1.2 g for 7 days, n : 7) or reloading (1.2 g for 4 days, n : 24) of amiodarone. All patients proved refractory to amiodarone alone. Nine VT were also refractory to endocardial catheter fulguration in 8 patients. Twenty one patients had coronary artery disease, 4 had arrhythmogenic right ventricular dysplasia, 4 had dilated cardiomyopathy, 1 had valvular disease, and 1 had no structural heart disease. Twelve patients had an ejection fraction less than 30 p. 100. Ten patients were in NYHA functional class 3. VT was permanent in 3 patients, daily in 5, weekly in 7, paroxysmal in 16. In 14 patients, VT occurred both at day and night. Oral administration of a daily low dose of a beta-blocker agent (acebutolol 100 mg, betaxolol 5-10 mg, metoprolol 50 100 mg, nadolol 20-40 mg, pindolol 2.5 mg, propranolol 30 mg, sotalol 80-160 mg, tertatolol 2.5 mg) combined with 400 mg per day of amiodarone suppressed VT episodes in all patients. None presented heart failure or collapse. The mean reduction of the heart rate was about 17 p. 100. One patient need a definite pacemaker to correct sinus bradycardia. At discharge, exercise ECG (n: 20) induced non sustained VT in 2 patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573327 TI - Molecular probes for the detection of Kluyveromyces marxianus chromosomal DNA in electrophoretic karyotypes of intergeneric protoplast fusion products. AB - Random genomic DNA fragments from Kluyveromyces marxianus were cloned in order to identify chromosomal bands in pulsed field electrophoresis patterns of intergneric hybrid strains which were obtained by protoplast fusion. Molecular hybridization data indicated that the K. marxianus parental strain might be triploid, and it showed strong chromosome length polymorphism. We analyzed the karyotype of two Saccharomyces cerevisiae/K. marxianus hybrid strains (St. 1.St.46) with our DNA probes and with a Ty1 specific probe. We found indications for recombinational events which lead to the formation of hybrid chromosomal DNA molecules. PMID- 2573328 TI - P3 in schizophrenia is affected by stimulus modality, response requirements, medication status, and negative symptoms. AB - Eighteen schizophrenics who were not taking medication, 13 schizophrenics who were taking medication, and 37 age-matched controls were tested with event related potential paradigms designed to elicit P3 response automatically or effortfully (ie, with a choice reaction time task). Electroencephalograms were recorded from the 19 standard 10-20 electrode sites. Compared with controls, both groups of schizophrenics had reduced P3 amplitudes for both effortful and automatic paradigms. P3 latencies were delayed relative to controls for the medication-taking schizophrenics in the effortful paradigms. Negative symptoms derived from the Brief Psychiatric Rating Scale within 1 week of event-related potential testing correlated negatively with both auditory and visual P3 amplitude in the subjects who were not taking medication. There was no evidence that P3 is smaller over left temporal electrode sites in schizophrenics, as has been reported by others. P3 amplitude reduction in schizophrenia is a robust psychobiological phenomenon that is present regardless of medication status or task demands. PMID- 2573329 TI - [Early stages of hepatic carcinogenesis after vagotomy]. AB - Initial stages of hepatocarcinogenesis have been studied in nonoperated and vagotomized animals. As a carcinogenic substance diethylnitrosamine (DENA) has been used. In order to estimate manifestation of the changes, the histochemical method for revealing glucoso-6-phosphatase activity, adenosine triphosphatase and gamma-glutamyltranspeptidase in the liver has been applied. The disturbance of vagus innervation is stated to delay the course of early stages of hepatocarcinogenesis, induced with DENA. PMID- 2573330 TI - [Morphofunctional evaluation of the testes in cryptorchism]. AB - In both abdominal and inguinal cryptorchism, complex morphological changes are observed with age in the undescended testis. Significant changes in the size of the seminiferous tubules are observed morphometrically; the most informative index was the surface of the tubuli and their relative volume in the total volume of the organ. The main importance in the development of cryptorchism belongs, in the authors' view, to the hormonal disturbance of the anterior lobe of hypophysis and Leydig cells which are practically absent in the undescended tests in fetuses, newborns and children of the first year of life with multiple congenital developmental abnormalities. PMID- 2573331 TI - Growth factors for neuronal survival and process regeneration. Implications in the mammalian central nervous system. AB - Within the past several years a number of substances have been identified in the mammalian brain that are capable of (1) preventing the death of injured neurons and (2) promoting the regeneration of severed neuronal processes. The goal of this review article is to update the clinical neurologist in this area by presenting a brief, general overview of this subject, including a glimpse at potential clinical implications. Recent advances in neuronal transplantation and molecular techniques underlying nerve growth are discussed. Possible therapeutic approaches are presented for many neurologic disorders, ranging from stroke to Alzheimer's disease to acquired immunodeficiency syndrome, based on regrowing or saving injured neurons. The clinical neurologist will become important in practical applications and research into prolonging neuronal survival and fostering axonal regeneration. Over the coming years, with further research, it is anticipated that patients will be treated with these or similar modulatory agents. PMID- 2573332 TI - Exploratory laparotomy alone in carcinoma of the oesophagus and gastric cardia. AB - A retrospective analysis of 453 patients with carcinoma of the oesophagus and gastric cardia was carried out in order to identify the incidence, operative findings, and outcome of patients who underwent laparotomy only without a definitive procedure. Of 343 patients who underwent surgery, 81% had their tumours resected and 15% had a bypass procedure. The remaining 14 patients (4%) had an exploratory laparotomy alone. The incidence of performing a laparotomy only was 1.5% for patients with carcinoma of the thoracic oesophagus but was 14% for patients with tumour of the gastric cardia (P less than 0.001). The reasons for exploration alone in these 14 patients were advanced local disease (12), bilobar liver metastases (seven), extensive abdominal lymph node metastases (seven), peritoneal seedlings (six) and malignant ascites (four). All except one patient had more than one feature which led to the decision of exploration only. As the overall incidence of exploratory laparotomy was low, it would not be appropriate for all patients to undergo exhaustive and expensive investigations. Surgical exploration continues to be the only reliable method to determine the actual extent of disease and whether a definitive procedure is possible. However, patients with cancer of the cardia with clinical features of advanced disease might warrant additional evaluations as the incidence of exploratory laparotomy alone with minimal prior investigation is relatively high. PMID- 2573333 TI - Investigations into the toxicity of Echium plantagineum in sheep. 1. Field grazing experiments. AB - A field grazing trial was undertaken to monitor the health and production of crossbred sheep grazing pasture where Echium plantagineum constituted a considerable proportion of the available forage. The trial, conducted for 19 months over successive grazing seasons, demonstrated a significant difference in production, with sheep on the E. plantagineum pasture being lighter and growing less wool compared with sheep on Echium-free pasture. No mortalities involving pyrrolizidine alkaloid poisoning were recorded in sheep grazing E. plantagineum, although there was histological evidence of moderately severe liver damage associated with high liver copper concentrations in at least one sheep following the grazing of large quantities of the plant. PMID- 2573334 TI - Investigations into the toxicity of Echium plantagineum in sheep. 2. Pen feeding experiments. AB - In a pen feeding trial fresh Echium plantagineum was fed as the sole diet to crossbred sheep with or without a history of previous access to the plant. Control groups received a diet of lucerne chaff and oats. During the trial, sheep on the Echium diet lost weight and deaths occurred with histological evidence of excessive copper accumulation, usually accompanied by pyrrolizidine alkaloid damage, in the liver and biochemical evidence of liver toxicity. It is concluded that E. plantagineum alone is not a suitable fodder for sheep and can be toxic due to its pyrrolizidine alkaloid content and high copper to molybdenum ratio. PMID- 2573335 TI - What the aircrew automated escape system and aircrew life support system equipment designers need from the investigating medical officer and pathologist. AB - A major problem apparent in many safety investigation reports concerning aviation mishaps, and especially in their component Flight Surgeon's Reports (FSRs), is the failure of the investigators to recognize needs beyond those of their immediate organizational structures and chains of command in conducting the investigation, and reporting the resultant facts and findings. If aircrew losses and serious injuries are to be reduced, other needs must also be considered and addressed. These additional needs include those of aircrew automated escape system (AAES) and aircrew life support system (ALSS) acquisition personnel who formulate, design, and test requirements, and AAES and ALSS designers and manufacturers who need to know how well and under what conditions of use their equipment is performing. Each mishap, in a sense, constitutes an extremely costly yet highly realistic test of the AAES and ALSS involved. If properly and thoroughly examined, these mishaps will yield exceptionally valuable insights into AAES and ALSS problems and successes and the reasons underlying system behavioral differences. This paper discusses a number of the AAES and ALSS community's needs which investigating medical officers should consider if the U.S. Navy is to gain from these expensive and often tragic mishaps. PMID- 2573336 TI - The effect of an aversive event on the return of fear. AB - An experiment on the return-of-fear (ROF) was carried out on 40 snake- or spider phobic subjects in order to determine whether an arousing event that occurs shortly before retest influences the magnitude of the ROF. Additionally, we attempted to produce a reinstatement of fear by introducing an unrelated aversive event (shock) after the fear had been reduced. The arousal manipulation successfully increased subjective arousal but not heartrate responsiveness. The increases in subjective arousal were not followed by increases in ROF, and the attempt to produce a reinstatement of fear did not succeed. A significant correlation between speed of fear-reduction and ROF was obtained. PMID- 2573337 TI - Cardiac and neuroendocrine responses to exposure therapy in height phobics: desynchrony within the 'physiological response system'. AB - Subjective, behavioral, cardiovascular and neuroendocrine responses were intensively monitored in two height phobics over a full course of exposure therapy and at follow-up. Both subjects showed rising cortisol responses and stable, non-extinguishing norepinephrine responses to height exposure over the course of treatment, while improvement occurred in subjective and behavioral response systems. They had differing heart rate responses. Despite desynchrony amongst anxiety response systems and within the physiological 'system' at treatment conclusion, both subjects had successful outcomes which were preserved at follow-up. Implications for the measurement of the 'physiological response system' in behavioral research are discussed. PMID- 2573338 TI - Effects of long-term xamoterol therapy on the left ventricular mechanical efficiency in patients with ischemic heart disease. AB - Myocardial oxygen uptake and an index of mechanical left ventricular efficiency were determined in basal conditions or during prolonged therapy with the new beta 1-adrenoceptor partial agonist xamoterol in 16 patients with mild to moderate ischemic heart failure. During xamoterol therapy, left ventricular end-diastolic pressure decreased from 24.4 +/- 6.5 to 17.8 +/- 8.6 mm Hg (P less than 0.01) and the isovolumic index of inotropic state (dP/dt)/DP40 increased by 14% (P less than 0.01). The heart rate increased slightly and the mean systolic and peak systolic wall stress also tended to increase (+ 7%; NS) but myocardial oxygen uptake (14.1 vs 14.7 ml/min; NS) and the index of efficiency (8.77 +/- 3.44 to 8.82 +/- 4.27; NS) were not significantly modified. In conclusion, prolonged therapy with xamoterol was not accompanied by a deterioration in the mechanical efficiency of the ventricle, even in patients with ischemic heart disease. PMID- 2573339 TI - Modulation of the autonomic control of the failing heart. AB - The failing heart operates with an abnormal combination of heart rate, stroke volume, and enddiastolic volume. This mismatch becomes more evident during exercise of patients with heart failure, when an increase in cardiac output is achieved with higher heart rate, a lower stroke volume and a higher enddiastolic volume. Using the beta 1-adrenoceptor partial agonist xamoterol which lacks beta 2-adrenoceptor agonism the response of the heart to sympathetic stimulation can be modulated. At rest and low levels of exercise xamoterol provides an inotropic support of the heart, whereas it reduces inappropriate tachycardia at higher levels. Thereby, xamoterol tends to normalize the balance of the inotropic and chronotropic control of the failing heart, because cardiac output is increased with a more normal combination of heart rate, stroke volume, and filling pressure. The beneficial effects of xamoterol are discussed as being especially important for failing ischemic hearts, because the balance between energy supply and energy demand may be improved by xamoterol. PMID- 2573340 TI - Dopexamine in congestive heart failure: how do the pharmacological activities translate into the clinical situation? AB - Dopexamine is a newly developed sympathetic catecholamine which combines dopaminergic (DA-1) and beta 2-adrenergic agonist activity with only minor beta 1 adrenergic action. Thereby, this compound exerts systemic and preferential renal vasodilation, causing afterload reduction, increases in cardiac output, and improved renal perfusion in animals and normal volunteers. Short-term administration of dopexamine in congestive heart failure established beneficial effects in central hemodynamics, that is, reduction of systemic and pulmonary vascular resistance, combined with a decrease in LV-filling pressures and increased renal blood flow. The effect on central hemodynamics are comparable to sodium nitroprusside. With higher doses, however, heart rate may increase substantially with dopexamine along with increased myocardial oxygen consumption. Experiences with prolonged administration of this drug are scarce and have, so far, yielded conflicting results. Thus, dopexamine appears to be a promising agent in the short-term management of congestive heart failure. However, its ultimate value for prolonged administration remains to be established. PMID- 2573341 TI - Clinical relevance of long-term therapy with levodopa and orally active dopamine analogues in patients with chronic congestive heart failure. AB - Beneficial effects of long-term treatment with dopamine analogues in patients with congestive heart failure may result from their vasodilating properties, in particular from renal artery vasodilation. Oral application of levodopa results in increased dopamine plasma levels and can improve cardiac performance and renal function in patients with congestive heart failure. A daily levodopa dosage of at least 4 g appears to a prerequisite for long-term response to the drug. Because of frequent side effects including nausea, vomiting, and dyskinesia at this dosage, the clinical usefulness of levodopa seems to be limited to a minority of patients. Ventricular arrhythmias have been shown to increase significantly during long-term levodopa therapy, probably due to stimulation of myocardial beta receptors. Increased ventricular arrhythmias or significant central nervous side effects have not been observed after administration of ibopamine and fenoldopam, which are orally active analogues of dopamine. Both agents exhibit potent arterial vasodilating properties and have been shown to increase cardiac performance in patients with congestive heart failure after short-term administration. The long-term beneficial effects of ibopamine and fenoldopam in the treatment of congestive heart failure have not yet been clarified. However, available results are encouraging and warrant further clinical evaluation of these agents, as well as the development of new analogues of dopamine, in particular of potent vascular dopamine agonists. PMID- 2573343 TI - [Sudden death in a drug milieu following use of an appetite suppressant]. AB - Chemical-toxicological analysis of blood and tissue samples from a drug-dependent person which had been found dead revealed the presence of propylhexedrine. The blood concentration was estimated to be 3.75 mg/l, this value may explain a fatal intoxication. However, death was obviously due to massive blood aspiration from nasal bleeding. PMID- 2573342 TI - On the mechanism of positive inotropic effects of alpha-adrenoceptor agonists. AB - The positive inotropic effect of the alpha 1-adrenoceptor agonist phenylephrine is accompanied by an increase in the presumed second messengers inositol 1,4,5 trisphosphate (1,4,5-IP3) and inositol 1,3,4,5-tetrakisphosphate (1,3,4,5-IP4). Both 1,4,5-IP3 and 1,3,4,5-IP4 sensitize myocardial contractile proteins in chemically skinned fibers. In addition to the Ca++ releasing effect of 1,4,5-IP3 from the sarcoplasmic reticulum the Ca++-sensitizing effect of the inositol phosphates may play a role in alpha 1-adrenergic positive inotropism. In isolated heart muscle preparations from patients with endstage heart failure (due to dilated cardiomyopathy) beta-adrenergic as well as alpha 1-adrenergic effects are reduced compared to preparations from healthy hearts. The reduced beta-adrenergic effects can in part be explained by an increased content of signal transducing G1 proteins. It is tempting to investigate whether other G proteins are also altered in severe congestive heart failure. PMID- 2573344 TI - [Glutamic acid group poisoning. So-called Chinese restaurant syndrome]. AB - After eating a soup 10 persons (out of 100) fell sick; within 10 minutes they suffered from nervous muscle convulsions, trembling, mouth desiccation and dilatation of the pupils. The soup contained glutamate as flavour enhancer in an unusually high concentration of 31 grams per litre. PMID- 2573345 TI - Treatment of rats with glucagon or mannoheptulose increases mitochondrial 3 hydroxy-3-methylglutaryl-CoA synthase activity and decreases succinyl-CoA content in liver. AB - 1. The activity of 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) synthase (EC 4.1.3.5) in extracts of rapidly frozen rat livers was doubled in animals treated in various ways to increase ketogenic flux. 2. Some 90% of the activity measured was mitochondrial, and changes in mitochondrial activity dominated changes in total enzyme activity. 3. The elevated HMG-CoA synthase activities persisted throughout the isolation of liver mitochondria. 4. Intramitochondrial succinyl-CoA content was lower in whole liver homogenates and in mitochondria isolated from animals treated with glucagon or mannoheptulose. 5. HMG-CoA synthase activity in mitochondria from both ox and rat liver was negatively correlated with intramitochondrial succinyl-CoA levels when these were manipulated artificially. Under these conditions, the differences between mitochondria from control and hormone-treated rats were abolished. 6. These findings show that glucagon can decrease intramitochondrial succinyl-CoA concentration, and that this in turn can regulate mitochondrial HMG-CoA synthase. They support the hypothesis that the formation of ketone bodies from acetyl-CoA may be regulated by the extent of succinylation of mitochondrial HMG-CoA synthase. PMID- 2573348 TI - Leukotriene metabolism and hypersensitivity in man and mouse. AB - The relative rates of cysteinyl-leukotriene metabolism were analyzed in fresh human and mouse serum. Human serum contained higher gamma-glutamyl-transpeptidase activity than mouse serum, and a higher percentage of the metabolized leukotriene C4 was recovered as leukotriene D4 in the human serum than in the mouse serum. The results suggest that the patterns of metabolism of the cysteinyl-leukotrienes could be an important factor in determining the relative sensitivity of an animal to the development of hypersensitivity reactions. PMID- 2573346 TI - Evidence for a role of dipeptidyl peptidase IV in fibronectin-mediated interactions of hepatocytes with extracellular matrix. AB - Dipeptidyl peptidase IV (DPP IV) is a cell surface glycoprotein which has been implicated in hepatocyte-extracellular matrix interactions [Hixson, DeLourdes, Ponce, Allison & Walborg (1984) Exp. Cell Res. 152, 402-414; Walborg, Tsuchida, Weeden, Thomas, Barrick, McEntire, Allison & Hixson (1985) Exp. Cell Res. 158, 509-518; Hanski, Huhle & Reutter (1985) Biol. Chem. Hoppe-Seyler 366, 1169-1176]. However, its proteolytic substrate(s) and/or binding protein(s) which mediate this influence have not been conclusively identified. Nitrocellulose binding assays using 125I-labelled DPP IV that was purified to homogeneity from rat hepatocytes revealed a direct interaction of DPP IV with fibronectin. Although fibronectin could mediate an indirect binding of DPP IV to collagen, no evidence was found for a direct binding of DPP IV to native or denatured Type I collagen. Fibronectin appeared to bind DPP IV at a site distinct from its exopeptidase substrate recognition site since protease inhibitors such as competitive peptide substrates and phenylmethanesulphonyl fluoride enhanced binding, possibly as a result of an altered conformation of DPP IV. To determine if fibronectin binding to DPP IV is involved in the interaction of fibronectin with the hepatocyte surface, the effect of various DPP IV inhibitors on 125I-fibronectin binding to isolated hepatocytes in suspension was examined. Kinetic studies revealed that inhibitors of DPP IV which enhanced fibronectin binding in vitro accelerated the initial binding of fibronectin to the cell surface where it was subsequently cross-linked (presumably by tissue transglutaminase) to as yet undefined components. Immunolocalization of fibronectin and DPP IV in normal rat liver sections showed that both proteins were present along the hepatocyte sinusoidal membrane. These observations, coupled with previous results showing that DPP IV is tightly bound to biomatrix isolated from rat liver (Hixson et al., 1984; Walborg et al., 1985), suggest that DPP IV binding to fibronectin may play a role in interactions of hepatocytes with extracellular matrix in vivo and possibly in matrix assembly. PMID- 2573347 TI - The presence of acyl-CoA hydrolase in rat brown-adipose-tissue peroxisomes. AB - The subcellular distribution of acyl-CoA hydrolase was studied in rat brown adipose tissue, with special emphasis on possible peroxisomal localization. Subcellular fractionation by sucrose-density-gradient centrifugation, followed by measurement of short-chain (propionyl-CoA) acyl-CoA hydrolase in the presence of NADH, resulted in two peaks of activity in the gradient: one peak corresponded to the distribution of cytochrome oxidase (mitochondrial marker enzyme), and another peak of activity coincided with the peroxisomal marker enzyme catalase. The distribution of the NADH-inhibited short-chain hydrolase activity fully resembled that of cytochrome oxidase. The substrate-specificity curve of the peroxisomal acyl-CoA hydrolase activity indicated the presence of a single enzyme exhibiting a broad substrate specificity, with maximal activity towards fatty acids with chain lengths of 3-12 carbon atoms. The mitochondrial acyl-CoA hydrolase substrate specificity, in contrast, indicated the presence of at least two acyl CoA hydrolases (of short- and medium-chain-length specificity). The peroxisomal acyl-CoA hydrolase activity was inhibited by CoA at low (microM) concentrations and by ATP at high concentrations (greater than 0.8 mM). In contrast with the mitochondrial short-chain hydrolase, the peroxisomal acyl-CoA hydrolase activity was not inhibited by NADH. PMID- 2573349 TI - Production and characterization of monoclonal antibodies against particulate guanylate cyclase in porcine kidney. AB - Three monoclonal antibodies (Ig G1 type) to particulate guanylate cyclase from porcine kidney cortex have been produced by fusing spleen cells from immunized BALB/c mouse with P3X63 myeloma cells. The antibodies were detected by their ability to bind immobilized antigen and by immunoprecipitation of enzyme activity. After subcloning by limiting dilution, hybridomas were injected intraperitoneally into mice to produce ascitic fluid. The antibodies recognized a 180,000 dalton protein in Lubrol-PX extract of porcine kidney cortex membrane, and when immobilized on Sepharose 4B, they co-precipitated both [125I]human atrial natriuretic peptide (ANP)-receptor complex and guanylate cyclase activity. The antibodies caused a greater increase in generation of cGMP than that of ANP. PMID- 2573350 TI - HeLa cells contain the atrial natriuretic peptide receptor with guanylate cyclase activity. AB - Receptors for atrial natriuretic peptide (ANP) are heterogeneous: an approximately 140-kDa receptor exhibits ANP-stimulated guanylate cyclase activity whereas an approximately 65-kDa receptor is thought to act only as a clearance storage protein. We have used photoaffinity labeling techniques to show that the human cell line, HeLa, contains predominantly the approximately 140-kDa ANP receptor. In contrast, several other cell lines contain primarily the approximately 65-kDa receptor. In HeLa cells, ANP bound specifically to high affinity binding sites (Kd approximately 2 nM) and stimulated a rapid, dose dependent accumulation of cGMP. These cell lines can thus provide useful models to study the multiple mechanisms of ANP action. PMID- 2573351 TI - Biosynthesis of endothelium-derived relaxing factor: a cytosolic enzyme in porcine aortic endothelial cells Ca2+-dependently converts L-arginine into an activator of soluble guanylyl cyclase. AB - In the presence of porcine aortic endothelial cytosol, soluble guanylyl cyclase purified from bovine lung was activated by L-arginine up to 2.5-fold, with an EC50 of about 6 microM. This activation was dependent on NADPH and Ca2+. The EC50 for Ca2+ was about 60 nM. No effect of L-arginine on guanylyl cyclase was observed when the cytosolic proteins were heat-denaturated. The effect of L arginine was inhibited by NG-monomethyl-L-arginine and hemoglobin. These results indicate that endothelial cells contain a cytosolic enzyme which is directly or indirectly regulated by Ca2+ and converts L-arginine into a compound which in stimulating soluble guanylyl cyclase behaves similar to endothelium-derived relaxing factor. PMID- 2573353 TI - Evidence for two separate beta-ketoacyl CoA reductase components of the hepatic microsomal fatty acid chain elongation system in the rat. AB - The hepatic microsomal fatty acid chain elongation system can utilize either NADPH or NADH. Elongation activity, measured as the rate of malonyl CoA incorporation into palmitoyl CoA, was enhanced by a fat-free diet and by bovine serum albumin (BSA) when either cofactor was employed. When the intermediate products were determined, it was observed that in the presence of BSA and NADPH, the predominant product was the saturated elongated fatty acid, whereas in the presence of BSA and NADH, the major intermediate was the beta-ketoacyl derivative. Employing beta-ketostearoyl CoA as substrate, BSA markedly inhibited NADH-supported beta-ketoacyl CoA reductase activity and stimulated NADPH supported activity. Furthermore, the sum of the NADH-dependent and NADPH dependent beta-ketoreductase activities approximated the activity obtained when both cofactors were present in the incubation medium, suggesting the existence of two beta-ketoacyl CoA reductases, one using NADH and the other, NADPH. PMID- 2573352 TI - Different gamma-glutamyl transpeptidase mRNAs are expressed in human liver and kidney. AB - In human, the two subunits of gamma-glutamyl transpeptidase (GGT) arise from a common precursor encoded by a multigene family. Until now, a single specific coding sequence for this precursor (type I) has been identified in human placenta and liver. In the present study, we have isolated from a human kidney cDNA library, a GGT specific clone (0.8 Kb). The sequence of which (type II) i) covers the carboxy terminal part of the GGT precursor, ii) exhibits 22 point mutations and a 30 bp deletion as compared to the type I GGT sequence. The sequencing of a human genomic clone reveals that this type II GGT mRNA is encoded by a different gene than the type I GGT mRNA. Both type I and type II GGT mRNAs are expressed in human liver, while almost exclusively type II GGT mRNA is detected in human kidney. PMID- 2573354 TI - Glutamine synthetase and glutaminase activities in various hepatoma cells. AB - Glutamine synthetase and glutaminase activities in a series of hepatoma cells of human and rat origins were determined for comparison with normal liver tissues. Marked decrease in glutamine synthetase activity was observed in the tumor cells. Phosphate-dependent and phosphate-independent glutaminase activities were increased compared with those from normal liver tissues. Well coupled mitochondria were isolated from HuH 13 line of human hepatoma cells and human liver. Oxypolarographic tests showed that glutamine oxidation was prominent in the tumor mitochondria, while mitochondria from the liver showed a feeble glutamine oxidation. Glutamine oxidation was inhibited by prior incubation of the mitochondria with DON (6-diazo-5-oxo-L-norleucine), which inhibited mitochondrial glutaminase. These results indicate that the product of glutamine hydrolysis, glutamate, is catabolized in the tumor mitochondria to supply ATP. PMID- 2573355 TI - Non-NMDA glutamate receptor antagonist 3H-CNOX binds with equal affinity to two agonist states of quisqualate receptors. AB - Binding of 3H-CNQX to rat cortical membranes is saturable and reversible. Apparently, 3H-CNQX binds to a single site with KD = 39 nM. However, studies using AMPA as inhibitor revealed a biphasic inhibition of 3H-CNQX binding. The results suggest that CNQX binds with the same affinity to two different sites. The molecular target size of 3H-CNQX binding (51.8 +/- 3.4 kD) is equivalent to the size of the high affinity 3H-AMPA binding sites, but different from the high affinity 3H-kainate binding sites. A monoexponential decay curve for the high energy radiation inactivation analysis of 3H-CNQX binding indicates that the two 3H-CNQX binding sites have the same molecular weight and therefore might be two different conformations of the same receptor. The standard excitatory amino acids quisqualate, AMPA and kainate have a different rank order of potency as binding inhibitors at the two conformations of the quisqualate receptor. PMID- 2573356 TI - Cell type specific, receptor-mediated modulation of growth kinetics in human lung cancer cell lines by nicotine and tobacco-related nitrosamines. AB - The objective of this study was to investigate a potential involvement of nicotinic cholinergic receptors in the mediation of cell type specific biological effects of nicotine and the two tobacco-related nitrosamines N nitrosodiethylamine (DEN) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) on human lung cells. Three well differentiated human lung cancer cell lines that have been reported previously to possess ultrastructural and biochemical features of normal pulmonary neuroendocrine cells, Clara cells and alveolar type II cells, respectively, were used for these experiments. The effects of nicotine, DEN, and NNK on cell proliferation and its modulation by established antagonists of nicotinic and muscarinic cholinergic receptors were studied. In the neuroendocrine cell line, nicotine and the two nitrosamines caused a strong stimulation of cell proliferation that was inhibited by antagonists of nicotinic cholinergic receptors. In the cell lines with features of Clara cells and alveolar type II cells, nicotine did not stimulate cell proliferation. Both nitrosamines stimulated cell proliferation in the cell line with Clara cell features. This effect was not changed by pre-exposure to cholinergic antagonists. The data suggest a selective uptake of nicotine and the two nitrosamines via nicotinic cholinergic receptors in pulmonary neuroendocrine cells. PMID- 2573357 TI - Mechanisms of multidrug resistance in HL60 cells. Analysis of resistance associated membrane proteins and levels of mdr gene expression. AB - HL60 cells isolated for resistance to Adriamycin do not contain P-glycoprotein, as determined with immunological probes. These cells, however, are multidrug resistant and defective in the cellular accumulation of drug. In view of these findings, we have examined in greater detail certain properties of the HL60/Adr cells and have compared these properties to an HL60 drug-resistant isolate (HL60/Vinc) which contains high levels of P-glycoprotein. The results of these studies demonstrated that verapamil induces a major increase in cellular drug accumulation in both HL60/Adr and HL60/Vinc isolates. An 125I-labeled photoaffinity analog of verapamil labeled P-glycoprotein contained in membranes of HL60/Vinc cells. In contrast, this agent did not label any protein selectively associated with drug resistance in membranes of the HL60/Adr isolate. The photoactive dihydropyridine calcium channel blocker [3H]azidopine and [125I]NASV, a photoaffinity analog of vinblastine, labelled P-glycoprotein in membranes from HL60/Vinc cells, whereas in experiments with the HL60/Adr isolate there was no detectable labeling of a drug resistance associated membrane protein. Additional studies have been carried out to analyze membrane proteins of HL60/Adr cells labeled with the photoaffinity agent 8-azido-alpha-[32P]ATP (AzATP32). The results demonstrate that this agent labeled a resistance associated membrane protein of 190 kilodaltons (P190). P190 is essentially absent in membranes of drug-sensitive cells. Labeling of P190 with AzATP32 in membranes of resistant cells was blocked completely when incubations were carried out in the presence of excess unlabeled ATP. Additional studies were carried out to analyze mdr gene amplification and expression in sensitive and resistant cells. Experiments carried out with human 5',mdr1 (1.1 kb) and mdr3 (1.0 kb) cDNAs demonstrate that both of these sequences were highly amplified in the HL60/Vinc isolate. Only the mrd1 gene sequence however, was overexpressed. In contrast, there was no detectable amplification or overexpression of mdr1 or mdr3 sequences in HL60/Adr cells. The results of this study thus identify a new nucleotide binding protein which is overexpressed in membranes of HL60 cells isolated for resistance to Adriamycin. P190, which exhibits properties distinct from P-glycoprotein, possibly functions in the energy-dependent drug efflux system contained in the HL60/Adr resistant isolate. PMID- 2573358 TI - [Disulfide bonds in neurotoxin-III from the sea anenome Radianthus macrodactylus]. AB - Positions of the three disulfide bridges in neurotoxin-III (RTX-III) from sea anemone Radianthus macrodactylus were determined: Cys3--Cys43, Cys5--Cys33, Cys26 -Cys44. The cystine-containing peptides obtained by the staphylococcal proteinase/trypsin digestion of the intact RTX-III were investigated. PMID- 2573359 TI - Rheumatoid arthritis complicated by infection with the human immunodeficiency virus and the development of Sjogren's syndrome. AB - A patient with seropositive rheumatoid arthritis became infected with the human immunodeficiency virus (HIV) through heterosexual transmission. After the primary phase of the HIV infection, persistent lymphadenopathy, leukopenia, and thrombocytopenia developed. Over the ensuing 3 years, the signs and symptoms of inflammatory polyarthritis completely disappeared, and severe Sjogren's syndrome developed. HIV and its associated immune dysfunction may be responsible for these events. PMID- 2573360 TI - The frequency of complement receptor type 1 (CR1) gene polymorphisms in nine families with multiple cases of systemic lupus erythematosus. AB - Low levels of complement receptor type 1 (CR1) on the erythrocytes of patients with systemic lupus erythematosus (SLE) may be acquired or genetically determined. Nine families with multiple cases of SLE were studied using a CR1 probe and restriction fragment length polymorphism analysis, to address this question. The absence of a significant increase in the frequency of a 6.9-kb band (previously shown to be associated with low-level CR1 on erythrocytes) suggests that this genetic marker does not play a major role in determining SLE, either in these families or in SLE patients in general. PMID- 2573361 TI - A new psychoactive 5H-2,3-benzodiazepine with a unique spectrum of activity. AB - The neuropharmacological effects of 1-(4-amino-phenyl)-4-methyl-7,8-dimethoxy-5H 2,3-benzodiazepine (GYKI 52 322) were investigated and compared with those of chlordiazepoxide and chlorpromazine. This novel 2,3-benzodiazepine displays neuroleptic activity in the apomorphine-climbing (ED50 = 1.15 mg/kg i.p.) and swim-induced grooming (ED50 = 6.9 mg/kg i.p.) tests in mice and it inhibits the conditioned avoidance response in rats (ED50 = 8.2 mg/kg i.p. and 9.8 mg/kg p.o.). However, it does not antagonize apomorphine-evoked vomiting in dogs; or stereotypy, hypermotility and turning in rats even at as high a dose as 50 mg/kg i.p. On the other hand it is active in the hole board test in mice (MED (minimal effective dose) = 0.5 mg/kg i.p.) and in the lick conflict assay in rats (MED = 5 mg/kg i.p.), indicating anxiolytic property. It shows antiaggressive effect in the fighting mice test (ED50 = 8.1 mg/kg p.o.) and the carbachol-rage procedure in cats (active at 10 mg/kg i.p.) According to the biochemical findings, this compound does not bind to the central dopamine receptors (IC50 greater than 10( 4) mol/l), but it shows affinity to the 5-HT1 receptors (IC50 = 7.1 x 10(-6) mol/l) and inhibits brain cAMP-phosphodiesterase (IC50 = 2.4 x 10(-5) mol/l). The substance causes no elevation of dopamine turnover and serum prolactin level suggesting fewer side effects. So the term "atypical neuroleptic agent" is proposed to characterize this molecule. PMID- 2573362 TI - Neurotransmitters in the brain. PMID- 2573363 TI - Five stars for a successful partnership. PMID- 2573364 TI - Early intervention in patients with excessive consumption of alcohol: a controlled study. AB - From a population of 2,114 patients attending somatic outpatient clinics, 78 patients were selected who had either an excessive consumption of alcohol according to questionnaires or a raised gamma glutamyltransferase (GGT) value (above 0.6 mu kat/l) due to alcohol. They had not undergone treatment for problem drinking previously, and had no serious alcohol dependence. They were thereby classified as excessive consumers of alcohol, and randomly allocated to an intervention (n = 36) or to a control group (n = 42). Those in the intervention group were followed up by a nurse once a month and by a doctor every third month for a total of 12 months. Laboratory tests were taken monthly. Consumption of alcohol, GGT and triglyceride levels, and sickness allowance days were decreased in the intervention group compared to the time before intervention. In contrast, the number of sickness allowance days in the control group increased. There was also a tendency towards a positive effect of intervention on the number of consultations made with a statistically not significant decrease of consultations after intervention. The study thus indicates that an early and relatively simple intervention programme for problem drinkers may be effective and can be carried out at a low cost and with a positive response from the patients. PMID- 2573365 TI - Beta-blockade and renal function in hypertension. Symposium. PMID- 2573367 TI - Pharmacological features of the vasodilation induced by tertatolol in isolated perfused rat kidneys. AB - The aim of the studies summarized in the present review was to obtain a pharmacological characterization of the in vitro renal vasodilator action of tertatolol. In isolated Tyrode-perfused rat kidneys, previously constricted with norepinephrine, serotonin or BaCl2, tertatolol evokes vasodilatation. These dilator responses cannot be explained by an interaction of tertatolol with alpha- or beta-adrenoceptors, muscarinic or nicotinic receptors, opioid receptors, dopamine receptors or histamine receptors, and they occur independently of the release of prostaglandins. Since methylene blue, an inhibitor of the soluble form of guanylate cyclase, reduces the renal dilator effect of tertatolol, this action could ultimately depend, at least in part, on the production of cyclic guanosine monophosphate; in this respect, the renal effects of tertatolol and atrial natriuretic factor (ANF) are different. From experiments performed with canine renal arteries and with the perfused rat mesentery, it can be concluded that the effect of tertatolol is more pronounced at the level of the resistance vessels. The in vitro renal dilator response observed with tertatolol may help to explain the beneficial effect on the renal circulation observed in both humans and experimental animals treated with the compound. PMID- 2573366 TI - Effect of tertatolol on glomerular hemodynamics in the normal Munich-Wistar rat. AB - Administration of beta-blockers is frequently associated with decreases in both renal plasma flow and glomerular filtration rate (GFR). Tertatolol, a new non selective beta-blocker, has demonstrated similar systemic effects to other conventional beta-blockers, but tertatolol produces increases in both renal plasma flow and glomerular filtration rate. To evaluate the renal hemodynamic mechanism responsible for the increase in GFR, micropuncture experiments were performed in normal Munich-Wistar rats. Intravenous injection of tertatolol produced a significant increase in GFR and urinary sodium excretion in spite of a decrease in systemic blood pressure. Single nephron glomerular filtration rate was significantly increased due to an increase in single nephron plasma flow related to parallel afferent and efferent arteriolar dilatation. No significant changes in glomerular hydrostatic pressure, transcapillary hydrostatic pressure gradient or the ultrafiltration coefficient were demonstrated with tertatolol. The capacity of tertatolol to increase glomerular filtration rate and to promote sodium excretion without modifying critical glomerular pressures makes this agent a highly attractive antihypertensive drug. PMID- 2573368 TI - Effects of tertatolol on renal function in the isolated perfused rat kidney. AB - Tertatolol, a new beta-adrenergic blocker, increases glomerular filtration rate (GFR) and renal plasma flow (RPF), and enhances diuresis and natriuresis in experimental animals and humans. The mechanism underlying the renal effects of tertatolol has not been established. In the present study we addressed the possibility that tertatolol influences renal function by a direct intrarenal mechanism. For this purpose we used a preparation of isolated rat kidney perfused with an artificial cell-free medium. Tertatolol when given as a bolus injection into the renal artery at the dose of 25 and 50 micrograms/kg, but not of 12.5 micrograms/kg, significantly enhanced the glomerular filtration rate (GFR) and perfusate flow rate (PFR). In contrast, the intrarenal bolus injection of different doses of propranolol (100, 250, or 500 micrograms/kg) was unable to change GFR and PFR to a significant extent. While no change in urine flow rate was found when the lowest dose of tertatolol was used, the compound at the dose of 25 and 50 micrograms/kg progressively increased urine flow during the time of perfusion. A similar effect of tertatolol was observed for urinary sodium and potassium excretion. In contrast, different doses of propranolol did not significantly change the urine flow rate or sodium and potassium excretion rates. We conclude that tertatolol, but not propranolol, increases GFR and PFR, and enhances urine output and sodium excretion in the isolated perfused rat kidney. These findings indicate that tertatolol preserves renal function by a mechanism independent of systemic changes. PMID- 2573369 TI - Alterations in renal hemodynamics during chronic and acute beta-blockade in humans. AB - Renal hemodynamics of hypertensive patients are characterized by an increased renal vascular resistance (RVR), resulting in low renal plasma flow (RPF). Normal glomerular filtration rate (GFR) is maintained through an increased filtration fraction (FF). This pattern may have long term harmful consequences for the kidneys. Propranolol, given in the short- or long-term, lowers both RPF and GFR. Pindolol, as well as so called cardioselective beta-blockers and labetalol, alter renal hemodynamics only slightly, if not at all. In some studies, nadolol has been shown to increase RPF, leaving GFR unchanged. A new beta-blocker, tertatolol, seems to consistently increase RPF and GFR. These different renal actions of various beta-blockers are poorly explained. They may be, however, of clinical significance, in the short term for patients with already impaired renal function, and possibly over the long term for other hypertensive patients. PMID- 2573370 TI - Antihypertensive action of beta-adrenoceptor blocking drugs. The role of intrarenal mechanisms. AB - The different modes of antihypertensive action of beta-adrenoceptor blocking drugs are reviewed with a special emphasis on renal mechanisms of action. Beta adrenoceptors occur in the kidney at the level of the juxtaglomerular and tubular cells. Direct intrarenal effects of beta-blockers include the decrease in renin release and the enhanced tubular excretion of fluid and electrolytes. Beta blockers also influence renal function in a more indirect way by altering the activity of different extrarenally formed hormones with an effect on fluid and electrolyte excretion, such as angiotensin II, atrial natriuretic factors and prostaglandins. Several beta-blockers have an additional effect on the renal vasculature. One of these, tertatolol, is a renal vasodilator beta-blocker in different species. The possible mechanisms of renal vasodilatation of tertatolol are reviewed. It is concluded that the shift of the renal pressure-diuresis curve is an essential feature in the mode of long-term antihypertensive action of beta blockers. PMID- 2573371 TI - Redistribution of cardiac output to the kidneys by tertatolol does not involve prostaglandins. AB - Renal perfusion has been shown to be preserved or improved during treatment by tertatolol in patients with arterial hypertension. The aims of the present study were (1) to document the central and renal hemodynamic effects of tertatolol in normal subjects and (2) to look for a possible interaction between tertatolol and products of the cyclooxygenase pathway of arachidonic acid metabolism. Five mg of tertatolol, 1 g aspirin, 5 mg tertatolol together with 1 g aspirin, and placebo were administered to 8 healthy volunteers at 1 week intervals in a random order and in a double-blind fashion. Cardiac output was measured by cardiac Doppler echography and renal blood flow and glomerular filtration rate by constant infusion techniques using I123-iodohippurate and Cr51-EDTA respectively. Measurements were performed before and then successively 2 and 4 h after oral intake of drugs or placebo. Tertatolol alone or with aspirin significantly decreased heart rate and cardiac output (P less than .05) without change in blood pressure, renal blood flow or glomerular filtration rate. The renal fraction of cardiac output was increased by tertatolol alone or with aspirin (P less than .05). Either placebo or aspirin alone had no effect. Thus tertatolol redistributes cardiac output to the kidneys in normal subjects as previously reported in hypertensive patients. This favorable effect on renal hemodynamics appers unlikely to be mediated by a local release of vasodilating prostaglandins. PMID- 2573372 TI - Structure-activity relationships as a response to the pharmacological differences in beta-receptor ligands. AB - With a few notable exceptions, beta-receptor ligands (agonists and antagonists) belong to the aryl- or heteroaryl-ethanolamine series and to the aryl- or heteroaryl-oxypropanolamine series. Structure-activity relationships for beta adrenergic agonists show that a secondary amine in the phenylethanolamine side chain ending is essential for receptor stimulation. The 3,4-dihydroxyphenyl groups may be replaced by "phenol equivalents" (-CH2OH, -NHCONH2, -CHOH, NHSO2CH3). In contrast, substitution at carbon alpha of the phenyl-ethanolamine side chain decreases or suppresses beta-adrenergic activity. The general requirements for beta-adrenergic blocking activity in the aryl- or heteroaryl oxypropanolamine are as follows: (1) the potency of beta-blockade is conferred by a branched alkyl group (isopropyl or tert-butyl) grafted on the terminal amino N, and by the nature and position of a substituent on the aromatic ring: ortho substituted compounds (especially when they have an hetero-atom in alpha) are the most potent ones. (2) The cardioselectivity is improved by the attachment of 3,4 dimethoxyphenylethyl,4-amide-substituted phenoxyethyl or acylamino-alkyl moieties to the terminal amino N of the side chain. Para substitution on the aromatic ring (particularly 4-acylamido substitution) has also yielded cardioselective drugs. Finally, the beta 1-selectivity is strongly and negatively correlated with lipophilicity. (3) Intrinsic sympathomimetic activity can be modulated by aromatic nucleus variations, particularly by hydroxyl-equivalents (electron withdrawing groups) on meta- and para-positions (3,4-substitutions). PMID- 2573373 TI - Reduction of beta-adrenergic receptors can explain the lack of rebound effect after tertatolol withdrawal. AB - Tertatolol is a potent beta-blocker with no intrinsic sympathomimetic activity (ISA) or beta 1/beta 2 receptor subtype selectivity. We provide evidence that tertatolol competitively inhibits beta-adrenergic receptors (beta-AR) and induces a marked and persistent reduction of their number. This has been consistently found in vitro and in vivo. The in vitro study showed that the receptor reduction by tertatolol was rapid (about 1 h at 37 degrees C), slowly reversible and independent of ISA. This effect was also observed in vivo. In healthy volunteers, seven days tertatolol treatment lowered the number of beta-AR by 26%. This number gradually rose back to the pretreatment levels, and a significant effect was still present on day 3 after drug withdrawal. The reduction of heart rate by tertatolol was also persistent and was still significant on day 3 to 5 after drug withdrawal. We conclude that the reduction of the receptor numbers may be important in producing a lack of a rebound effect after discontinuation of chronic tertatolol treatment. PMID- 2573374 TI - Tertatolol in chronic renal failure. A pharmacokinetic study. AB - Pharmacokinetics of tertatolol were investigated in 22 hypertensive patients (12 men and 10 women; mean age +/- SD: 52.6 +/- 12.3 years) with chronic renal failure defined by a mean creatinine clearance (Clcr) of 24.6 +/- 15.9 mL/min/1.73 m2 (range: 6.2 to 68.7). A daily single dose of 5 mg tertatolol was administered orally for 4 weeks, except in the 72 h following the first administration. Plasma samples and urine collections were carried out over 72 h after the first (D0) and the last dose (D27). After the first administration, tertatolol was rapidly absorbed (time to peak concentration: 1.2 +/- 0.7 h) and peak concentration was 160 +/- 80 ng/mL. Plasma concentrations decreased following a biphasic curve, with two half-lives of 2.5 +/- 1.1 and 17.0 +/- 8.5 h, respectively. These parameters were not modified by repeated administration of tertatolol and did not significantly correlate with Clcr either at D0 or at D27. Plasma levels were stable along the study with similar areas under plasma curves following the first and the last dose (P = NS). In addition, plasma levels extrapolated from first dose data did not significantly differ from those observed during repeated dosage. Plasma levels of the 4-OH metabolite which possesses a beta-blocking activity were low, inconstantly detectable, not related to the degree of renal impairment, and no accumulation occurred after chronic dosage. Renal excretion of tertatolol and 4-OH tertatolol was significantly increased by repeated administration (P less than .01) and correlated well with Clcr either at D0 or at D27. Four week treatment was well tolerated and significantly improved Clcr (+6.5%, P less than .02). In conclusion, tertatolol was well tolerated and did not accumulate in patients with renal failure of various degrees. The usual daily single dose of 5 mg may be kept unchanged whatever the degree of renal impairment. PMID- 2573375 TI - Cardioprotection and antihypertensive therapy. Are beta-blockers the only effective therapy? AB - Hypertension is a major risk factor for stroke and coronary heart disease and this increases with increasing age. It is a chronic disease which affects a quarter of the adult population. On its own or in conjunction with other risk factors, especially cigarette smoking, hypertension is associated with serious morbidity and high mortality. A large number of clinical trials have been undertaken to assess the efficacy of antihypertensive therapy in controlling high blood pressure and in reducing the morbidity and mortality associated with hypertension. These trials have studied more than 40,000 subjects worldwide as a result of which the benefits of treatment even in milder forms of hypertension are well established today. However, antihypertensive therapy has been disappointingly unsuccessful in the primary prevention of coronary artery disease despite the undoubted protection against malignant phase hypertension, heart failure, stroke and renal failure as demonstrated in the early Veterans Administration trial and subsequent studies. The success of post-infarction beta blockade suggested that the wider use of beta-blockers prophylactically might limit hypertension-related coronary disease. Taking into account the adverse and metabolic effects of thiazide diuretics, beta-blockers together with calcium channel blockers must remain the drugs of choice as step one therapy in most patients with hypertension. PMID- 2573376 TI - Effects of long-term antihypertensive treatment with tertatolol on diastolic function in hypertensive patients with and without left ventricular hypertrophy. AB - We assessed the effects of long-term antihypertensive treatment with 5 mg tertatolol, a noncardioselective beta-blocker, on left ventricular hypertrophy (LVH) and diastolic function. Fifteen hypertensive patients were selected as good responders to previous treatment with tertatolol (supine blood pressure less than 140/90 mm Hg). They were divided into 2 groups: group 1 with LVH (n = 6) and group 2 without LVH (n = 9). After a one month wash-out period, all patients received 5 mg tertatolol once daily. In case of uncontrolled blood pressure (BP), the dose was doubled after 2 weeks in 10 patients. BP control was obtained in all patients. M-mode echocardiography and Doppler-echocardiography were performed under controlled conditions after BP normalization and after 6 months of treatment. Long-term BP normalization significantly reduced left ventricular mass index (LVMI) in group 1 (from 137 +/- 3 to 121 +/- 3 g/m2, P less than .01), but not in group 2 (from 120 +/- 3 to 114 +/- 4 g/m2, P = NS). After 2 weeks of effective therapy, the ratio between early and late diastolic peak flow velocity across the mitral valve (E/A ratio), significantly increased in both groups (from 0.72 +/- 0.04 to 0.87 +/- 0.06 in group 1, P less than .05; and from 1.13 +/- 0.06 to 1.26 +/- 0.07 in group 2, P less than .05). After 6 months, together with the reduction of LVMI, a further increase of E/A ratio was only observed in group 1 (to 1.30 +/- 0.12, P less than .05). IN CONCLUSION: (1) LVH contributes to left ventricular diastolic dysfunction in hypertensive patients since its reversal is able to improve diastolic filling, and (2) effective antihypertensive treatment with tertatolol improves diastolic function independently from its effect on LV mass. PMID- 2573377 TI - Antihypertensive effects of tertatolol. A comparative study versus atenolol. AB - The purpose of this study was to compare the antihypertensive efficacy and tolerance of tertatolol and atenolol. Sixty-one ambulatory patients with uncomplicated mild to moderate hypertension (diastolic blood pressure (DBP) between 95 and 120 mm Hg) received, following a randomized allocation, either 5 mg tertatolol (T) (n = 30) or 100 mg atenolol (A) (n = 31) in single daily dose. After a one-month single-blind placebo run-in period, the two drugs were administered double-blind over a 3 month period. Blood pressure (BP) and heart rate were measured before, and after the first and third months. Electrocardiographic and biological data were collected respectively before and at the third month. At each consultation, patients were asked to report any side effects. Atenolol and tertatolol were similar in controlling BP: the decrease in supine systolic blood pressure (SBP) reached 24.2 mm Hg (P less than .001) in group T and 21.7 mm Hg in group A (P less than .001), and the decrease in DBP reached 16.6 mm Hg in group T (P less than .001) and 16.8 mm Hg in group A (P less than .001). Supine heart rate (HR) fell by 15.5 beats/min and 14.8 beats/min in the T and A groups, respectively (P less than .001). At the end of the trial, blood pressure control (DBP less than or equal to 90 mm Hg) was obtained in 80% and 70% of the patients in the T and A groups, respectively. The clinical tolerance of both drugs was satisfactory with 8 slight and transient complaints in T group, and 14 complaints with one treatment-related withdrawal at the second month in A group. Electrocardiographic and biochemical variations were not significantly different in the two groups. We conclude that the antihypertensive efficacy of 5 mg tertatolol is comparable to that of 100 mg atenolol, with a similar clinical and biochemical tolerance. PMID- 2573378 TI - Tertatolol in hypertension. Long-term therapy in 2,338 patients. AB - Tertatolol has been studied in 2,338 patients with mild to moderate hypertension over a one year period. Tertatolol (T) was initiated alone (5 mg once daily) and if satisfactory control of blood pressure (BP) (diastolic BP less than 95 mm Hg) was not achieved, treatment was adapted at the third or sixth month either by increasing the dosage (heart rate greater than 70 beats/min), or by adding a potassium-sparing diuretic (heart rate less than or equal to 70 beats/min). Blood pressure normalization was achieved in 88.8% of the study population: 66.1% on single and 22.7% on dual therapy. The decrease of diastolic BP was 18.4 mm Hg (from 102.8 +/- 0.2 to 84.4 +/- 0.2 mm Hg, P less than .001). Tertatolol alone or associated with diuretic (T + D) induced a significant and continuous decrease in supine systolic and diastolic BP. Overall side effects were rare, leading in only 6.5% of the cases to the discontinuation of the drug. Plasma creatinine significantly decreased in the single therapy group only (from 92.2 +/- 0.5 to 90.1 +/- 0.5 mumol/L, P less than .01). In patients with initial plasma creatinine greater than or equal to 100 mumol/L (n = 661), plasma creatinine markedly decreased (-10%, from 114.6 +/- 0.7 to 103.4 +/- 0.8 mumol/L, P less than .01), and to the same extent with T or T + D. Thus, this large-scale study confirms that tertatolol is an efficient and well-tolerated antihypertensive drug, which improves renal function, especially when initially reduced. PMID- 2573379 TI - The Tertatolol International Multicenter Study (T.I.M.S.). Predicting factors of an efficient therapy. AB - The Tertatolol International Multicenter Study (T.I.M.S.) was aimed at assessing the efficacy and tolerability of 5 mg tertatolol (T) once daily over a one year period and at finding, among the clinical variables before treatment, parameters predicting a successful single beta-blocking therapy. The study was carried out in 230 patients (96 men, 134 women, mean age +/- standard error of the mean: (51.5 +/- 0.7 years) suffering from uncomplicated mild to moderate hypertension. After a one-month placebo run-in period (M0-M1) and a one-month double-blind placebo-controlled period (M1-M2), 213 patients entered and 166 completed the one year open period (M2-M14). A diuretic (D) was added if blood pressure (BP) was inadequately controlled with the single therapy. After one year, 88.5% of patients were controlled (sitting diastolic BP less than or equal to 90 mm Hg): 56.3% on single therapy (T) and 32.2% on dual therapy (T + D), respectively. Statistical analysis performed on the controlled subjects showed that patients who were controlled with T alone had an initially lower standing diastolic BP and a higher standing heart rate, were younger and had a lower sitting systolic BP than those controlled with T + D. Using such discriminant parameters which are predictors, 83.5% patients (single therapy group) and 49% patients (dual therapy group) were classified in the therapeutic group to which they really belonged. This study confirms the antihypertensive efficacy of tertatolol, and highlights relevant parameters for a successful single beta-blocker therapy in clinical practice. PMID- 2573380 TI - Intracranial abscesses secondary to nasal, sinus, and orbital infections in adults and children. AB - Nineteen cases of intracranial abscesses secondary to infection of the midface are reported. The most common underlying cause was bacterial sinusitis. Other etiologic factors included mucormycosis following steroid therapy, Wegener's granulomatosis, nasal dermoid cyst and sinus tract, tooth abscesses, aspergillosis following chemotherapy for leukemia, squamous cell carcinoma of the frontal sinus, infected methylmethacrylate plate for a prior skull fracture, and a case of gauze packing left in the sinus following surgery. Anaerobic organisms were the predominant cause of the abscesses. The most dangerous intracranial complication was subdural abscess, which occurred in seven patients in this series. Three of them died. Four cases of frontal and parietal lobe abscesses were treated with systemic antibiotics only. This approach has not been well emphasized in our literature. Steroid therapy should not be used for the treatment of sinus and orbital infections. It can result in dreadful complications. The overall mortality rate in this series was 21% (4 of 19), despite aggressive treatment and close cooperation between the neurosurgeon, otolaryngologist, and other specialists. Early diagnosis and adequate treatment are paramount. PMID- 2573381 TI - Prognostic indicators in head and neck cancer patients receiving combined therapy. AB - In 1983 we initiated a prospective nonrandomized study of the value of preoperative chemotherapy in previously untreated patients with stages III and IV squamous cell carcinoma of the head and neck. In 1983 and 1984, 50 patients were entered in the study. Prior to therapy all patients were evaluated by a representative from the Medical Oncology, Radiation Therapy, and Head and Neck Surgery Divisions, University of Utah School of Medicine, Salt Lake City. In addition to the standard preoperative evaluation, pretreatment computed tomographic scans were performed on all patients. Follow-up computed tomographic scans were performed after the second cycle of chemotherapy and at the completion of treatment. Initial therapy in all patients consisted of induction chemotherapy with cisplatin (day 1, 100 mg/m2) and fluorouracil (days 1 through 5, 1000 mg/m2). Several factors were examined for their utility in predicting response to therapy and survival. Factors evaluated included: (1) extent and timing of chemotherapeutic response; (2) computed tomographic quantitated primary tumor size; (3) size of computed tomographic quantitated regional (neck) metastases; (4) performance status; (5) cancer stage; (6) total lymphocyte count; and (7) serum liver function tests. The factor found to be most useful in predicting improved survival was the extent of response to chemotherapy. The remaining factors, performance status, regional lymph node status, serum gamma glutamyltransferase levels, and cancer stage, were also found to correlate with length of survival but were much less important than the response to chemotherapy. PMID- 2573382 TI - Neurones and neuropeptides in coelenterates. AB - The first nervous system probably evolved in coelenterates. Many neurons in coelenterates have morphological characteristics of both sensory and motor neurones, and appear to be multifunctional. Using immunocytochemistry with antisera to the sequence Arg-Phe-NH2 (RFamide), RFamide-like peptides were demonstrated in the nervous systems of all classes of coelenterates. Using a radioimmunoassay for RFamide, three such peptides were isolated from the sea anemone Anthopleura elegantissisma and sequenced: less than Glu-Gly-Arg-Phe-NH2 (Antho-RFamide), less than Glu-Ser-Leu-Arg-Trp-NH2 (Antho-RWamide I) and less than Glu-Gly-Leu-Arg-Trp-NH2 (Antho-RWamide II). The general structure of these peptides can be described as less than Glu...Arg-X-NH2, where X is an aromatic amino acid. From the hydromedusa Polyorchis penicillatus, the peptide less than Glu-Leu-Leu-Gly-Gly-Arg-Phe-NH2 (Pol-RFamide I) was isolated, which also belongs to the less than Glu...Arg-X-NH2 family. Using specific antisera it was shown that all four peptides were located in neurones. Application of low doses of Antho-RFamide, or Antho-RWamide I or II induced contractions of endodermal muscles of sea anemones. This indicates that these neuropeptides play a role in neurotransmission. PMID- 2573383 TI - Phylogenetic considerations of neurosecretory granule contents: role of nucleotides and basic hormone/transmitter packaging mechanisms. AB - The characteristics of neurosecretory granules include the presence of an acidic interior, a hyperosmolar concentration of granule solutes, the presence of chromogranin (CG) or CG-like soluble acidic proteins and a high content of nucleotides, predominantly ATP. The identification of "nucleotides" within the neuroendocrine "stem cells" of coelenterates (e.g. Hydra) has raised some interesting evolutionary questions as to the function of intragranular nucleotides. The chromaffin granules of adrenal medullary cells have been studied extensively, and are representative of the neurohormone/neurotransmitter packaging problems encountered in neurosecretory granules, in general. At the acid pH (5.7) of the interior of the chromaffin granule, ATP has three negative charges based on the pK value of the gamma-phosphate group. ATP can therefore interact with positively charged amines, acetylcholine and divalent cations, forming binary and ternary complexes. The results of nuclear magnetic resonance (NMR) spectroscopy indicate that the hyperosmolar solutes within the chromaffin granule exist in a viscous, but fluid state; one function of ATP could be to help lower the osmotic pressure of the granule contents through extensive, but weak, intermolecular bonding. In addition, ATP is an excellent buffer to help maintain a pH of 5.7 within the interior of the chromaffin granule. An acidic milieu contributes to neurohormone/neurotransmitter packaging and granule stability. The presence of nucleotides within neurosecretory granules cannot, however, be explained on the basis of the ability of ATP to simply reduce osmotic pressure, since insulin molecules exist in a crystalline phase, a condition which, by itself, could substantially reduce osmotic pressure; nucleotides, nevertheless, co-exist in these insulin cores. ATP and ATP metabolites such as ADP, AMP and adenosine, formed as a result of the action of ectonucleotidases, can have extensive extracellular trophic and feedback effects after secretion. Extracellular nucleotides and adenosine can function as neuromodulators, agonists and antagonists to inflammatory cells, and regulators of blood flow, etc. It is possible that intragranular nucleotides were retained through a billion or more years of evolution because of the importance of these trophic and feedback effects. Parts of the neuroscretory granule, such as the F1 subunit of the proton translocating ATPase, can be traced back to the aerobic bacteria, vacuolar amine transport to yeast and a CG-like acidic protein to protozoan secretory granules (i.e., the trichocysts of Paramecia). PMID- 2573384 TI - The hypothalamo-hypophyseal complex: a paradigm of the paraneuron concept. AB - The neuronal and epithelial parts of the hypothalamo-hypophyseal complex (proximal and distal neurohypophysis, adenohypophysis) have in common the task of message transfer. According to the paraneuron concept, neurons and endocrine cells as paraneurons produce similar or identical substances, store them in vesicles and release them in response to specific stimuli. The released substances reach their targets via the intercellular spaces in a paracrine mode or via the blood in a more or less far-reaching hemocrine way. Special contacts for message transfer are not mandatory, even in the case of neurons. The biological activity of substances released by neurons and paraneurons depends on their affinities to binding sites, most probably localized on the outer target cell membrane. The neurons and paraneurons under discussion are mostly localized close to the mid-sagittal plane. The common ontogenetic roots of some neurons and paraneurons might be a further important point of the paraneuron concept. PMID- 2573385 TI - Pharmacokinetics of sufentanil in neurosurgical patients undergoing hyperventilation. AB - The pharmacokinetics of sufentanil were determined in 11 hypocapnic (arterial PCO2 2.9-3.7 kPa) and seven normocapnic (arterial PCO2 5.3-6.0 kPa) neurosurgical patients. Following a single i.v. bolus of sufentanil 4 micrograms kg-1, multiple arterial samples were obtained at timed intervals and plasma concentration of sufentanil was measured by radioimmunoassay. Calculation of the pharmacokinetic parameters from the derived compartmental models demonstrated an increased total volume of distribution of sufentanil in hypocapnic patients (mean 5427 (SD 1866) ml kg-1 v. 3518 (1097) ml kg-1; P less than 0.05) in the control patients and a more prolonged elimination half-life (232 (60) min v. 143 (51) min; P less than 0.01). The increased distribution of sufentanil with hyperventilation was probably caused by an increased proportion of opioid in the nonionized state. PMID- 2573386 TI - Relative potency of vecuronium on the diaphragm and the adductor pollicis. AB - To quantify the neuromuscular blocking effect of vecuronium on the diaphragm and the adductor pollicis, single twitch stimuli were applied to the phrenic nerves at the neck and the ulnar nerve at the wrist in anaesthetized patients. The evoked responses were measured simultaneously by recording the transdiaphragmatic pressure with a differential pressure transducer and the adductor pollicis strength with a force displacement transducer. Cumulative vecuronium dose response curves were determined for both muscles in 11 ASA class I adult patients. The mean (SD) doses required to depress adductor pollicis and diaphragm responses to 50% (ED50) were 30 (9) micrograms kg-1 and 37 (12) micrograms kg-1, respectively. Corresponding values for 95% depression of the twitch response (ED95) were 48 (13) micrograms kg-1 and 67 (23) micrograms kg-1 (P less than 0.02), indicating that the diaphragm required approximately 40% more vecuronium for subtotal abolition of the single twitch response. PMID- 2573387 TI - Effects of a new neuromuscular blocking agent (Org 9426) in anaesthetized cats and pigs and in isolated nerve-muscle preparations. AB - The effects of Org 9426 (the 2-morpholino, 3-hydroxy, 16N-allyl pyrrolidino analogue of vecuronium) were studied in anaesthetized cats and pigs and in isolated nerve--muscle preparations using tension and intracellular recording techniques. In isolated preparations, the effects of Org 9426 were antagonized by neostigmine. No contracture of the chick muscle preparation occurred. Org 9426 reduced the amplitude of endplate currents (EPC) in rat and snake muscle, but had no major effects on EPC decay characteristics, indicating a lack of endplate channel blocking action. In anaesthetized animals, no fasciculations were observed and the neuromuscular block was associated with tetanic and train-of four fade and was antagonized by neostigmine. In anaesthetized cats and pigs, Org 9426 was approximately 20% as potent as vecuronium, its onset of action was twice as rapid as that of vecuronium in the cat and its duration of action was similar to that of vecuronium in both cats and pigs. It blocked the bradycardia produced by vagal stimulation only in doses greater than those necessary to produce neuromuscular block (ratios 7.2 in the cat and 4.4 in the pig--10-14% of the corresponding ratios for vecuronium). Ganglion block was seen only at doses several times those producing vagal block. In general the effects of Org 9426 on the cardiovascular system were slight, a small depressor effect occurring at high doses in the cat. The 17-hydroxy analogue, the potential metabolite of Org 9426, was approximately 20 times less potent than Org 9426 and is thus unlikely to contribute to the neuromuscular block produced by the parent compound. PMID- 2573388 TI - Sensitivity to both vecuronium and neostigmine in a sero-negative myasthenic patient. AB - We describe a sero-negative ocular myasthenic patient who showed exaggerated responses to both vecuronium and neostigmine. These hypersensitive responses were not anticipated because preoperative clinical and laboratory evaluations suggested a negligible impairment of somatic muscles by myasthenic processes. This case report re-emphasizes that, if neuromuscular blocking agents and their antagonists are administered to myasthenic patients, regardless of the degree of impairment, the dose should be titrated carefully with the aid of a neuromuscular transmission monitor. PMID- 2573389 TI - The natural history of human immunodeficiency virus infection in a haemophilic cohort. AB - 112 haemophilic patients infected with HIV were followed up with clinical and laboratory assessment between 1 December 1979 and 30 November 1988. Sixty-six (59%) of the patients developed HIV-related clinical symptoms and 22 (20%) developed AIDS. Twenty (18%) of the patients developed p24 antigenaemia. Amongst the 59 patients whose date of seroconversion could be estimated the calculated 8 year cumulative incidence of AIDS was 40% (symptoms 73%). For the whole cohort of 112 patients, the median slope of linear regression of the absolute T4 lymphocyte count was steeper for those with AIDS (-0.113 x 10(9)/l per year) than for those without AIDS (-0.054 x 10(9)/l per year) (P less than 0.02). While 15 cases of AIDS developed during 58 patient-years of follow up after falling below a T4 lymphocyte count of 0.2 x 10(9)/l, only two cases occurred during 450 patient years before reaching this count. Thus the decline of the T4 lymphocyte count to 0.2 x 10(9)/l may be an appropriate additional end-point for the assessment of new treatments for asymptomatic patients infected with HIV. PMID- 2573390 TI - cDNA cloning and sequence and cDNA-directed expression of human P450 IIB1: identification of a normal and two variant cDNAs derived from the CYP2B locus on chromosome 19 and differential expression of the IIB mRNAs in human liver. AB - A cDNA designated hIIB1, representing the entire coding sequence of a P450 in the IIB gene family, was isolated from a human liver lambda gt11 library by using the rat IIB1 cDNA as a probe. The hIIB1 protein, deduced from the cDNA sequence, contained 491 amino acids, had a calculated molecular weight of 56,286, and displayed 76% amino acid similarity with the rat IIB1 protein. Expression of this cDNA, using the vaccinia virus system, yielded a P450 that had a reduced CO binding spectrum with an absorption maximum of 452 nm. The expressed human enzyme was able to catalyze the deethylation of 7-ethoxy-coumarin. Total RNA from 13 livers was probed for levels of hIIB mRNA. Two livers had high levels, four contained moderate levels, and eight contained very low, or no detectable, mRNA. These data suggest either that defective hIIB1 genes exist in humans or that the hIIB1 gene is regulated and variably induced in our liver specimens. To search for mutant mRNA transcripts, libraries were constructed from livers expressing low levels of hIIB1 mRNA. A cDNA, designated hIIB2, was isolated that was identical with the hIIB1 cDNA except for the presence of an unusual alteration of the DNA near its 5' end corresponding to the putative exon 4. This alteration was caused by a deletion of 29 bp and an insertion of 44 bp of nonhomologous DNA. This sequence replacement occurs at the junction of the third and fourth exons as predicted from the structure of the rat IIB1 gene, suggesting that a faulty splice might have given rise to the variant hIIb2 transcript.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573391 TI - Simultaneous synthesis and degradation of glutamine in isolated rat liver cells. Effect of vasopressin. AB - When hepatocytes suspensions obtained from whole livers of 48-h-fasted rats were incubated in Krebs-Henseleit buffer with a near-physiological concentration (1 mM) of L-[1-14C]glutamine as substrate, the apparent removal of glutamine was low, but the release of 14CO2 was much larger than the enzymatically measured removal of glutamine. This indicates that glutamine was metabolized at rates much higher than those accounted for by the apparent removal of glutamine. This also suggests that glutamine utilization was, at least in part, masked by concomitant synthesis of glutamine from endogenous substrates via glutamine synthetase. Evidence that such synthesis occurred was obtained by: (i) addition of methionine sulfoximine, an inhibitor of glutamine synthetase, which caused a large increase in the apparent removal of glutamine; and (ii) measurement of the specific radioactivity of L-[1-14C]glutamine which was shown to decrease during incubation. Addition of vasopressin (10(-7) M) led to a marked increase in glutamine removal by a dual mechanism: it accelerated flux through glutaminase, the enzyme which initiates the hepatic degradation of glutamine, and inhibited flux through glutamine synthetase. PMID- 2573392 TI - Cyclic GMP and nitroprusside inhibit the activation of human platelets by fluoroaluminate. AB - Sodium nitroprusside, an activator of the soluble guanylate cyclase, inhibits the intracellular Ca2+ mobilization, ATP secretion and aggregation of human platelets evoked by fluoroaluminate. Similar results are obtained with 8-bromo-cyclic GMP (8-Br-cGMP). Both nitroprusside and 8-Br-cGMP inhibit the protein kinase C dependent phosphorylation of the 47 and 20 kDa proteins induced by fluoroaluminate, but not by the protein kinase C activators phorbol ester and diacylglycerol. Since fluoroaluminate interacts directly with a G protein, the present results suggest that the cGMP interferes with platelet activation at the level of G protein-phospholipase C. PMID- 2573393 TI - The cardiac sodium channel shows a regular substate pattern indicating synchronized activity of several ion pathways instead of one. AB - Cardiac sodium channel substates were induced by using different gating modifiers, namely S-DPI 201-106 (s), toxin II from Anemonia sulcata (a), veratridine (v) and mixtures of these agents (s + v, a + v). Current ratios (normalized substate currents), slope conductances, reversal potentials and saturation characteristics were evaluated for the individual channel substates. The results can be summarized as follows: (i) Current ratios fell into a pattern of six equidistant values (I to VI) irrespective of the modification applied (0.20, 0.34, 0.51, 0.69, 0.85, 1.00). Slope conductances, determinable for substates II, V and VI (4.8, 11.7 and 14.0, respectively), are also consistent with six conductance substates which are integer multiples of a smallest conductance (state I). (ii) The permeability ratio PNa+/PK+ (i.e., reversal potential of substate currents) of the sodium channel was conserved both for different modifications, i.e., by s, a, s + v and a + v, and for the different substates (at least for II, IV and VI) observed for each modification. (iii) Sodium binding to the channel is substate independent. Analysis of slope conductances of states II and VI for three sodium chloride concentrations (71.5, 140 and 303 mM) revealed different maximal conductances (geVImax = 2.9.geIImax) but similar apparent affinities for sodium (KNa + VI = 286 mM; KNa + II = 303 mM). These findings are shown to seriously challenge the commonly unquestioned conception that 'single-current events' reflect ion passage through only one single pathway. The alternative view, that not one pore, but either six or three pores with synchronized gating ('oligochannel') underlie 'single-channel events', is shown to readily account for the observed substate properties and appears not to contradict known properties of 'the sodium channel'. This fundamentally new view of the sodium channel aims to invoke further efforts to distinguish between conceptually distinct models of structure-function relationships for a variety of channels which show multiple substates and conserved ion selectivity. PMID- 2573394 TI - [Synaptosomal factors, regulating the activity of Na,K-ATPase and its sensitivity to neurotransmitters]. AB - The factors regulating the activity of synaptosomal Na,K-ATPase have been found in nerve endings cytosol. One of these (Mr 10,000) essentially inhibits, whereas the other one (Mr 60,000) in the presence of norepinephrine, 5-hydroxytryptamine and dopamine activates the Na,K-ATPase of synaptosomes. Regardless of their direct action on Na,K-ATPase, the effect of the activating factor increases in the presence of neurotransmitters. In cell ksap obtained by microsome precipitation the activating factor is absent. PMID- 2573395 TI - [Oncogenes and anti-oncogenes in breast cancer]. AB - Recent developments in molecular oncology have permitted the definition of alterations in proto-oncogenes (ras, myc, neu and hst/int.2) and anti-oncogenes (Rb.1) in human breast carcinomas. Detection of these abnormalities could be of prognostic interest. However, this remains controversial. PMID- 2573396 TI - Phototransduction in cones: an inverse problem in enzyme kinetics. AB - Phototransduction is a process which links the absorption of photons by a rod or cone to the modulation of voltage across the cell membrane. An important feature of many vertebrate photoreceptors is a mechanism that adjusts the sensitivity and dynamics of the response to light according to the level of illumination. We construct a system of ordinary differential equations that models what are currently thought to be the important molecule mechanisms involved in phototransduction: this includes consideration of both intracellular enzyme kinetics and the properties of light-insensitive and light-sensitive conductances in the cone membrane. The system contains negative feedback whose functional form is determined by constraining the steady-state behaviour of the system. Despite the highly nonlinear nature of the system of ordinary differential equations, our methods permit us to derive an analytic expression for the first-order frequency response parametric in the steady-state value of only one dynamic variable, the light input. Various unknown kinetic parameters are found by fitting the model to experimental data on the first-order frequency response of cones measured at several mean light levels spanning a range of four log units. Good fits are obtained to the data, and the computed shape of the feedback function agrees qualitatively with recent experiment. Moreover, the model accounts for the dramatic speeding up of the response kinetics and the decrease in response gain with increasing light level. PMID- 2573397 TI - Allogeneic transplantation of blood-derived, T cell-depleted hemopoietic stem cells after myeloablative treatment in a patient with acute lymphoblastic leukemia. AB - This report describes an allogeneic peripheral blood stem cell transplant in a patient who had received marrow ablative therapy. The patient was an 18-year-old white male with acute lymphocytic leukemia in third remission for whom an allogeneic bone marrow transplant was recommended. His HLA-identical sibling preferred to donate peripheral blood stem cells rather than marrow. The donor cells were collected with 10 apheresis procedures and depleted of T lymphocytes to prevent excessive graft-versus-host disease. Nine collections were cryopreserved. The patient received high-dose cytosine arabinoside and 12 Gy of total body irradiation, followed by infusion of all cryopreserved donor cells. A portion of the tenth apheresis product collected on the day of transplant containing 1.8 x 10(9) T lymphocytes was infused without further processing to approximate the number of T lymphocytes given in an allogeneic bone marrow transplant; the remainder was T lymphocyte depleted and infused. More than 1 x 10(9)/l granulocytes were present on day +11. A bone marrow biopsy on day +27 showed trilineage engraftment. Cytogenetic studies demonstrated that the recipient's marrow and peripheral blood were populated exclusively with donor cells. Allogeneic peripheral stem cell transplantation produced an early hematopoietic engraftment. Since the patient died on day +32, sustained engraftment could not be evaluated. PMID- 2573399 TI - Restriction fragment length polymorphism analysis of hematopoietic cells following successful treatment of relapsed acute lymphoblastic leukemia following bone marrow transplantation. AB - Despite aggressive therapy for leukemia in the form of bone marrow transplantation (BMT) relapse occurs in a significant number of cases. The origin of the leukemic relapse, whether it is of donor or recipient origin, and how best to treat the patients continue to pose problems for the clinician. In this paper we present a case in which the cytogenetics suggested that the relapse was of donor origin; however, molecular analysis revealed that the leukemic population was of host origin. The leukemic relapse following the BMT was treated with a second BMT. This resulted in a remission of 28 months after which leukemic relapse was again diagnosed. Using conventional chemotherapy it was possible to obtain another complete remission. This case illustrates a pitfall to cytogenetic analysis and two contrasting methods of dealing with leukemic relapse following BMT. PMID- 2573398 TI - CD8+/DR+/CD25--T-lymphocytes associated with marrow graft failure. AB - Phenotypic and functional characteristics of peripheral blood mononuclear cells (PBMC) were studied in eight patients with poor graft function following HLA identical T cell-depleted marrow transplantation. Similar patients with good graft function and normal individuals were used as controls. Freshly isolated PBMC from patients with failing grafts contained more CD3+ and CD8+ cells than PBMC from well engrafted patients. The CD8+ cells appeared activated insofar as they expressed DR antigens, but they did not express the low affinity IL-2 receptor recognized by Tac antibody (CD25) and they did not have increased cytolytic activities. After culture with phytohemagglutinin (PHA) and IL-2, PBMC from patients with poor graft function contained fewer CD2+ and CD4+ cells than cultured PBMC from patients with good graft function. Cultured cells from patients with poor graft function acquired lymphokine activated killer (LAK) activity against NK-sensitive and NK-insensitive targets, but still did not express CD25. Host-mediated anti-donor cytotoxic activity could be demonstrated in one patient only after presensitization with donor cells and culture with IL-2 and PHA. The abnormalities in T cell activation observed in patients with poor graft function did not correlate with the donor or host origin of lymphoid cells. These data indicate that some cases of graft failure may be associated with defective T cell maturation. These abnormalities may simply represent a consequence of marrow failure or they may actually contribute to failure by not providing critical hematopoietic accessory functions. PMID- 2573400 TI - A comparison of haemoglobin and erythrocytes as inhibitors of smooth muscle relaxation by the NANC transmitter in the BRP and rat anococcygeus and by EDRF in the rabbit aortic strip. AB - 1. The inhibitory effect of erythrocyte suspensions and haemoglobin solutions on the response of the bovine retractor penis muscle (BRP) and the rat anococcygeus to field stimulation of their non-adrenergic non-cholinergic (NANC) nerves has been compared. Haemoglobin 3 microM greatly reduced the relaxant response in both tissues whereas a haemoglobin-equivalent suspension of erythrocytes was without effect. 2. A similar comparison of erythrocytes and haemoglobin on the response of the rabbit aortic strip to EDRF liberated by acetylcholine (ACh) showed that both reduced EDRF-mediated relaxation, though haemoglobin was significantly more effective. 3. These results suggest that the NANC transmitter may not be as freely diffusible through the erythrocyte membrane as EDRF and may therefore not be nitric oxide. PMID- 2573401 TI - Lorazepam discontinuation promotes 'inverse agonist' effects of benzodiazepines. AB - 1. The effects of lorazepam discontinuation on responses to benzodiazepine agonists and antagonists were studied in mice. 2. The convulsant dose of pentylenetetrazol was decreased after an acute dose of lorazepam (0.5 mg kg-1) at 4 days after drug discontinuation, compared to 1 or 7 days after discontinuation or to vehicle treatment. 3. The percentage of mice undergoing convulsions after an acute dose of FG 7142 (40 mg kg-1) was increased at 4 days after lorazepam discontinuation, compared to 1 or 7 days after discontinuation or to vehicle treatment. 4. After an acute dose (0.5 mg kg-1), lorazepam concentrations in cortex tended to be greater in lorazepam-treated compared to vehicle-treated mice at 4 days after discontinuation compared to 1 and 7 days. 5. These data indicate a shift toward reduced agonist sensitivity and increased inverse agonist sensitivity in mice 4 days after lorazepam discontinuation. PMID- 2573402 TI - Demonstration in Tupaia papillary muscle preparations of alpha-adrenoceptors mediating positive inotropic effects: comparison with guinea-pigs. AB - 1. Positive inotropic responses to alpha- and beta-adrenoceptor agonists of isolated papillary muscles from the tree shrew (Tupaia), were compared with those from guinea-pigs. 2. In Tupaia, the concentration-response curve for phenylephrine, unlike that for isoprenaline, was not affected by pindolol in a concentration (10(-8) M) sufficient to block beta-adrenoceptor-mediated responses, but it was significantly shifted to the right by phentolamine (10(-6) M). In guinea-pig papillary muscles, however, the concentration-response curve for phenylephrine, like that for isoprenaline, was shifted to the right by pindolol (10(-8) M) but was unaltered by phentolamine (10(-6) M). Furthermore, when the mean concentrations of agonists inducing maximal positive inotropic responses were compared (relative to that of isoprenaline = 1.0), phenylephrine was found to be only slightly less potent (0.84 +/- 0.04; n = 5) in Tupaia and much less potent (0.33 +/- 0.06; n = 5) in the guinea-pig. 3. Although in Tupaia papillary muscles the increase in developed tension induced by a combination of phenylephrine and isoprenaline did not significantly differ from that by phenylephrine alone, it was approximately 3 times larger than that produced by phenylephrine alone in guinea-pigs. 4. These results indicate that in papillary muscles from Tupaia, unlike the guinea-pig, the positive inotropic effects of phenylephrine can be mediated by alpha-adrenoceptors. PMID- 2573403 TI - Simultaneous measurement of endothelium-derived relaxing factor by bioassay and guanylate cyclase stimulation. AB - 1. Endothelium-derived relaxing factor (EDRF) released by cultured endothelial cells (EC) from bovine aortae was measured by bioassay using pre-contracted strips of rabbit aorta and by radioimmunoassay of guanosine 3':5'-cyclic monophosphate (cyclic GMP) produced by stimulation of bovine lung soluble guanylate cyclase. 2. Bradykinin (Bk, 3 and 30 pmol) injected through a column of EC caused release of EDRF as detected by bioassay and increased cyclic GMP concentrations. Superoxide dismutase (SOD, 15 u ml-1) increased the amount of EDRF detected by the activation of soluble guanylate cyclase. 3. In the absence of endothelial cells, nitric oxide (NO, 1-2 microM), arachidonic acid (AA, 3-30 microM) or sodium nitroprusside (SNP, 1-100 microM) stimulated guanylate cyclase. Superoxide dismutase strongly increased the stimulation of guanylate cyclase induced by NO, but had little effect on the stimulation induced by SNP and no effect on the stimulation induced by AA. 4. Oxyhaemoglobin (10-300 microM) abolished the stimulation of guanylate cyclase by EDRF, NO or SNP but was much less effective as an inhibitor of AA-induced stimulation of guanylate cyclase. 5. These results demonstrate that measurement of guanylate cyclase stimulation by radioimmunoassay is a viable method for detecting EDRF release, especially useful when the drugs used interfere with bioassay tissues. PMID- 2573404 TI - Facilitatory effects of opioids on the discharges of visceral nociceptors. AB - Effects of opioids on the activity of visceral nociceptors were tested using the in vitro testis-spermatic nerve preparations excised from deeply anesthetized dogs. Morphine, DADLE and dynorphin (10 microM) elicited discharges of polymodal receptors in approximately 1/3 of the tested cases. The incidence of the excitatory response of morphine increased at higher concentrations. The excitatory responses were quite variable among preparations and showed a strong tendency for tachyphylaxis. Similar increases in discharges were elicited when morphine was applied during the steady state of the response evoked by bradykinin (BK) or BK mixed with prostaglandin E2. Pretreatment of morphine for 5 min significantly augmented the subsequent BK responses for 30 min or more. Naloxone per se induced neither excitation nor augmentation of the subsequent BK response, however it reversed the augmenting effect of morphine on BK response. In contrast with previous reports proposing peripheral analgesic effects of opioids, suppressive effects on nociceptors were never observed in the present experiment. Peripheral effects of opioids were discussed. PMID- 2573405 TI - Morphological, neurochemical, and behavioral characterizations associated with the combined treatment of diethyldithiocarbamate and 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine in mice. AB - Changes in striatal dopamine (DA) neurochemistry, tyrosine hydroxylase immunocytochemistry of DA fibers, and behavior following the combined administration of diethyldithiocarbamate (DDC) and 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) to mice were assessed. The combined treatment of DDC and MPTP produced a dose-dependent decrease in striatal DA levels and a dose related increase in the striatal DOPAC:DA ratio. Cumulative doses of MPTP equal to or exceeding 53.0 (26.5 mg/kg x 2. i.p.), given in combination with DDC, were effective in reducing striatal DA levels to less than 25% of control levels 2 weeks after treatment. Tyrosine hydroxylase immunocytochemistry revealed large deafferentation of DA terminal regions in striatum, moderate reductions in nucleus accumbens and dendritic regions of substantia nigra, and slight reductions in the number of DA cell bodies in substantia nigra. Mice treated with DDC and MPTP became hyperactive during the light phase of their diurnal cycle: psychopharmacological data suggest that postsynaptic DA receptors were supersensitized following this treatment. These data provide evidence that the combined treatment of DDC and MPTP produces severe and enduring depletion of mesostriatal DA, and also concomitant behavioral changes in mice. PMID- 2573406 TI - Somatostatin hyperpolarizes neurons and inhibits spontaneous activity in the rat dorsolateral septal nucleus. AB - Intracellular recordings were made from rat brain neurons in a submerged slice preparation containing the dorsolateral septal nucleus (DLSN). Somatostatin-14 (SS-14) was applied to these neurons by superfusing solutions containing known concentrations of the peptide or by pressure ejection from micropipettes. With either method of treatment, SS-14 produced membrane hyperpolarization and decreased membrane resistance in a concentration-dependent manner. The hyperpolarizing response to SS-14 occurred in virtually all neurons tested and appeared to result from a direct action on DLSN neurons mediated by an increased permeability to potassium ions. The SS-14-induced membrane hyperpolarization was not blocked by naloxone, bicuculline, tetrodotoxin, or calcium-free, high magnesium superfusion media. In a small number of neurons, SS-14 application produced a membrane depolarization which did not exhibit clear concentration dependence and was blocked by superfusion of calcium-free, high-magnesium media indicating an indirect action. These findings reveal that SS-14 is a potent inhibitor of DLSN neurons in vitro and provide the first evidence that receptors for this putative neurotransmitter are located on postsynaptic neurons in this nucleus. Synaptically released SS-14 may play an important role in the modulation of septohippocampal function. PMID- 2573407 TI - The dendritic dopamine projection of the substantia nigra: phenotypic denominator of weaver gene action in hetero- and homozygosity. AB - While cerebellar granule cell migration and survival are affected by the weaver (wv) mutation both in the heterozygous and homozygous states, the dopamine (DA) deficit of the nigrostriatal projection has been shown to involve only midbrain DA cell bodies and nigrostriatal DA axons of homozygous mutants. We have identified a cellular site which is defective in the mesencephalic DA system of mice both heterozygous and homozygous for the wv gene. That deficit involves the dendritic DA projection which extends from the substantia nigra pars compacta (SNc) into the pars reticulata (SNr). In the midbrain of heterozygotes, dopaminergic dendrites are reduced by 60% at 20 days of age, when DA neurone number in the midbrain, DA content in the neostriatum and pattern of synaptic connectivity of nigrostriatal axon terminals are normal. At the same age, the deficit of dopaminergic dendrites in the SNr of homozygotes (76%) is disproportionate to the loss of DA cell bodies (42%). These findings: (a) may provide clues to the aetiopathogenetic mechanisms of wv gene operation; and (b) may explain the generalised convulsions intermittently manifested by weaver heterozygotes, as the SN has been implicated in the pathophysiology of experimental seizures. PMID- 2573408 TI - Monoamine deficiency in a transgenic (Hprt-) mouse model of Lesch-Nyhan syndrome. AB - The integrity of forebrain monoamine systems has been assessed both biochemically and immunohistochemically in transgenic mice carrying the mutant hprt-bm2 gene, an animal model of Lesch-Nyhan syndrome. The mutant mice manifested 20-30% depletions of forebrain dopamine, and corresponding increases in dopamine turnover. By contrast, the mutant mice manifested normal tyrosine hydroxylase immunostaining of catecholamine cell bodies and terminals throughout the forebrain, and cell counts revealed no detectable loss of ventral mesencephalic dopamine neurones. Serotonin concentrations were also depleted, whereas no significant changes were found in noradrenaline or adrenaline, methylhydroxyphenylglycol (MHPG) or 5-hydroxyindoleacetic acid. The results indicate that a primary genetic deficiency in purine salvage pathways is associated with additional changes in forebrain monoamine metabolism in mouse as in man, although these changes are less pronounced in the animal model than in the human syndrome. The biochemical changes were not associated with explicit degeneration of the associated populations of neurones. PMID- 2573409 TI - Cultured astroglia release a neuronotoxic activity that is not related to the excitotoxins. AB - Neuronal death after brain injury is thought to be in part the result of the activity of the excitotoxins, a family of excitatory amino acids which are released by neurones. We have also described an astroglial cell-derived neuronotoxic activity of low molecular weight whose release can be induced by depolarizing events such as an increase in extracellular potassium concentration. We study here the relationship between this astroglia-derived neuronotoxic activity present in astroglia-conditioned medium (ACM) and the excitotoxins. Using a colorimetric assay of neuronal survival, we show that the ACM neuronotoxic activity, is able to induce the death of all types of neurones tested, including those which are insensitive to excitotoxins. Furthermore, the ACM neuronotoxic activity does not require for its action the extracellular ionic composition which is needed for the activity of excitotoxins. Finally, the ACM neuronotoxic activity is not blocked by competitive or non-competitive antagonists of the various classes of excitotoxin receptors. Those data demonstrate that the astroglia-derived neuronotoxic activity is not related to the excitotoxins. Still, because astrocytes can also be depolarized by members of the excitotoxin family, the possibility exists that the release of astroglia derived neuronotoxic activity would follow the rise in extracellular excitatory amino acid concentration during nervous system injury. PMID- 2573410 TI - Cholinergic differentiation of clonal rat pheochromocytoma cells (PC12) induced by retinoic acid: increase of choline acetyltransferase activity and decrease of tyrosine hydroxylase activity. AB - The effects of retinoic acid (RA), a naturally occurring metabolite of vitamin A, on the growth, morphology and neurochemical differentiation of the PC12 clone of rat pheochromocytoma cells were investigated. RA added to the medium inhibited the growth of PC12 cells in a dose-dependent manner up to 10 microM without affecting their morphology. In PC12 cells cultured in the presence of 10 microM RA for 8 days, the specific activity of choline acetyltransferase (ChAT) was increased 2-fold, while the specific activity of tyrosine hydroxylase (TH) was decreased 0.5-fold compared with cells cultured in the absence of RA. Specific activities of acetylcholinesterase (AChE), glutamate decarboxylase (GAD) and lactate dehydrogenase (LDH) were not affected by RA. Both the increase of ChAT and the decrease of TH induced by RA exhibited similar time and dose dependencies. RA inhibited the increase of TH activity induced by nerve growth factor (NGF), an adrenergic neuronotrophic factor on PC12 cells. From these observations it was concluded that RA induces a cholinergic neurochemical differentiation of PC12 cells independent of a morphological differentiation. PMID- 2573411 TI - An ultrastructural study of dynorphin-immunoreactive nerve fibers and terminals in the celiac-superior mesenteric ganglion of the guinea pig. AB - Dynorphin-immunoreactive nerve fibers and terminals were identified in the celiac superior mesenteric ganglion of the guinea pig at the ultrastructural level with the peroxidase-antiperoxidase technique. The immunostained material was localized in the large dense core vesicles of the terminals but was also present diffusely in the axoplasm. The terminals formed numerous axodendritic and a few axosomatic contacts, interpreted as synapses, with the principal ganglion cells. These findings suggest that dynorphin plays a role as a neurotransmitter or neuromodulator in the ganglion and, taken together with earlier findings, indicate an involvement of dynorphin neurons in the intestino-intestinal inhibitory reflex. PMID- 2573412 TI - The self-selected walking pace test and beta blockade. AB - The self-selected walking pace test is a safe, simple and inexpensive test to measure an individual's perception of walking speed and is more strongly related to maximal oxygen uptake than to age. This study was carried out to determine whether beta blockade, which commonly causes fatigue, would affect the test. Twenty-three clinically healthy men with a mean age of 43 years completed the test both on and off 80 mg long-acting oxprenolol daily, a beta blocking drug with partial agonist activity. The men walked 80 m at three self-selected speeds (rather slow, normal and rather fast). There was a significant reduction in heart rate on the drug. The walking speed at the three paces on and off medication were 1.08 ms-1 and 1.07 ms-1 (slow), 1.32 ms-1 and 1.33 ms-1 (normal) and 1.63 ms-1 and 1.61 ms-1 (fast). None of these differences reached statistical significance. The stride length also did not change. It is concluded that beta blocking medication does not interfere with the self-selected walking pace test. PMID- 2573413 TI - [The law of informed consent and its impasses in psychiatry]. AB - Every medical intervention is submitted to the rule of informed consent. Over the years, criteria of consent validity and exception situations have been defined. After a discussion of the difficult application of the informed consent rule in psychiatry, this article suggests an analysis of the motivations of a refusal of neuroleptic medication in 20 psychotic patients of a psychiatric hospital. The irrational motivations of refusal (particularly, denial and delusional ideation) have been evoked much more often then rational motivations (therapeutic inefficiency, secondary effects). The authors question the denial as a sufficient reason to declare a psychotic patient incompetent to consent. The consequences of the refusal of the neuroleptic treatment in some patients, mainly the risk of criminalization, are discussed. PMID- 2573414 TI - Amplification of protooncogenes in surgical specimens of human lung carcinomas. AB - The possible existence of amplification or rearrangement of protooncogenes was examined in more than 100 surgical specimens of human lung carcinoma. Protooncogenes were amplified in 28% of the carcinomas. About 90% of the amplified genes were of the myc, ras, or erbB family. Of the myc family genes, myc was amplified in 14 of 137 tumors and L-myc in four of 108 tumors, but N-myc was not amplified. A high frequency of amplification of myc was observed in squamous cell carcinomas (seven of 37) and of L-myc in small cell carcinomas (two of six). Of the ras family genes, K-ras-2 was amplified in six of the 137 tumors and N-ras in two of the 137 tumors, but no amplification of H-ras-1 was detected. Seven of the eight cases of amplified ras genes were in advanced pathological stages. Of the erbB family genes, erbB-1 (epidermal growth factor receptor) was amplified in 10 of 114 tumors and erbB-2 (HER-2/neu) in one of 51 tumors. Amplifications of the myc, ras, and erbB family genes might be one of the crucial DNA abnormalities involved in the development of human lung carcinomas. PMID- 2573415 TI - Changes in estrogen receptor, DNA ploidy, and estrogen metabolism in rat hepatocytes during a two-stage model for hepatocarcinogenesis using 17 alpha ethinylestradiol as the promoting agent. AB - 17 alpha-Ethylestradiol (EE2) was administered chronically to diethylnitrosamine (DEN)-initiated (200/mg/kg, i.p.) adult ovariectomized Sprague-Dawley rats, by means of Silastic implants at an estimated dose of 90 micrograms/kg/day. Isolated hepatocytes from DEN/EE2-treated animals exhibited a 2- to 3-fold increase in nuclear estrogen receptor (ER) levels throughout the promotion period. Furthermore, approximately 30-40% of the receptor was occupied when quantified by an exchange assay. For all groups the ER had a sedimentation coefficient of approximately 8S for unoccupied ER and a binding affinity for 17 beta-estradiol of 0.25 nM. An ER of lower affinity for estradiol was present in animals initiated with DEN and/or promoted with EE2. The increase in hepatocyte ER was associated with a 5.2-fold increase in gamma-glutamyl transpeptidase and 2.5-fold decrease in glucose-6-phosphatase activity at 20 weeks. EE2 treatment caused a 50% increase in the maximal binding capacity (Bmax) of hepatic epidermal growth factor receptors, but the equilibrium binding constant (Kd) did not change. Modulation of mitotic activity of hepatocyte subpopulations by EE2 treatment was indicated by an increase in the proportion of diploid hepatocytes and an increase in the number of hepatocytes undergoing DNA synthesis. In general, effects on ER, epidermal growth factor receptor, gamma-glutamyl transpeptidase and glucose-6 phosphatase were greater in DEN/EE2-treated animals than in rats receiving only EE2. Modification of receptor pathways associated with hepatocyte growth control, ER and epidermal growth factor receptor, may be contributing factors in the clonal expansion of preneoplastic cells during EE2 promotion of hepatocarcinogenesis. PMID- 2573417 TI - Loss of heterozygosity for loci on chromosome 17p in human malignant astrocytoma. AB - Loss of constitutional heterozygosity for specific chromosomal loci, when found consistently in a particular tumor type, suggests that a recessive oncogene important in the genesis of that tumor may be present within the involved chromosomal loci. DNA markers that detect restriction fragment length polymorphisms are powerful tools that have been used to detect loss of chromosomal loci in a growing number of human malignancies. The human brain tumor astrocytoma is usually malignant and virtually incurable. Two types of malignant astrocytomas are recognized histopathologically:anaplastic astrocytoma and glioblastoma multiforme. We carried out a restriction fragment length polymorphism analysis of tumors from 15 patients with anaplastic astrocytoma and 20 patients with glioblastoma using polymorphic DNA markers for loci on chromosome 17. Loss of constitutional heterozygosity for loci on chromosome 17 was found in both anaplastic astrocytoma and glioblastoma patients with equal frequency (40% of cases). Our mapping data revealed a region of loss on chromosome 17p between physical loci p11.2 and pter that was common to both patient groups. Taken together with the previously reported finding of loss of heterozygosity for loci on chromosome 10 in glioblastoma, these results indicate that tumorigenesis in the astrocyte lineage may involve recessive oncogenes on two different chromosomes. PMID- 2573416 TI - Multidrug resistance in a human leukemic cell line selected for resistance to trimetrexate. AB - Trimetrexate (TMQ) is a lipophilic antifolate shown to have antitumor activity in humans. TMQ-resistant sublines of the MOLT-3 human acute lymphoblastic leukemia cell line were developed and were designated as MOLT-3/TMQ200, MOLT-3/TMQ800, and MOLT-3/TMQ2500 based on degrees of resistance to TMQ. The TMQ resistance was accompanied by 5- to 7-fold increases in dihydrofolate reductase activity and markedly reduced cellular TMQ accumulation. Methotrexate accumulation was not impaired in TMQ-resistant cells. TMQ retention (efflux) was unchanged in these TMQ-resistant cells. Verapamil enhanced the TMQ accumulation in the resistant cells to the level seen in the parent cells but had no effects on the TMQ retention. These sublines were cross-resistant not only to methotrexate but also to vincristine, doxorubicin, daunorubicin, and mitoxantrone. There was no cross resistance to bleomycin or cisplatin. Resistance to vincristine, doxorubicin, daunorubicin, and mitoxantrone was reversed by verapamil. TMQ resistance was only minimally reversed by verapamil and methotrexate resistance not affected at all. Both cellular accumulation and retention of vincristine and daunorubicin in the TMQ-resistant cells were markedly decreased. Verapamil enhanced both accumulation and retention of the drug. Plasma membrane fractions of the TMQ-resistant cells analyzed by urea-sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by staining with Coomassie Blue revealed the presence of a distinct band with a molecular weight of 170,000. Immunoblot analysis with 125I-labeled monoclonal antibody raised against P-glycoprotein of multidrug-resistant Chinese hamster ovary cells (C219) cross-reacted with the Mr 170,000 protein of the TMQ resistant cells. These results show that the TMQ-resistant cells displayed not only decreased TMQ uptake and increased dihydrofolate reductase but also characteristics associated with a classical multidrug-resistant phenotype. Multidrug resistance includes lipophilic antifolate. PMID- 2573418 TI - Immunomodulation by various nitrosoureas and its effect on the survival of the murine host bearing a syngeneic tumor. AB - Chemotherapeutic efficacies of the nitrosoureas 1,3-bis(2-chloroethyl)-1 nitrosourea (BCNU), chlorozotocin (CLZ), and streptozotocin (STZ) were investigated against the LSA tumor which is syngeneic to C57BL/6 mice. It was observed that a single injection of 20 mg/kg body weight of BCNU or CLZ, even at an advanced stage of tumor growth, completely cured greater than 90% of the tumor bearing mice. Furthermore, BCNU-cured or CLZ-cured mice could specifically reject secondary rechallenge with the LSA tumor. In contrast, a single dose treatment with STZ at 20-200 mg/kg body weight failed to cure the tumor-bearing mice (0% survival). The failure of STZ to cure tumor-bearing mice was next addressed considering three possible mechanisms: (a) STZ was less tumoricidal; (b) STZ suppressed the immunity of the host; and (c) STZ failed to eliminate tumor specific suppressor T-cells. The failure of STZ to cure tumor-bearing mice was not totally related to its tumoricidal properties since STZ at higher doses did possess significant tumoricidal activity in vitro and in vivo, comparable to that of BCNU or CLZ. When spleen cells from normal mice treated with BCNU, CLZ, or STZ were assayed for their responsiveness to the T-cell mitogens concanavalin A or phytohemagglutinin, it was observed that STZ was in fact less immunosuppressive than BCNU or CLZ. The fact that STZ did not suppress the immunity of the host was also suggested by the findings that BCNU-cured mice treated with STZ or CLZ could still reject secondary rechallenge with the specific tumor LSA. Following treatment of tumor-bearing mice with BCNU or CLZ, tumor-specific delayed type hypersensitivity responses were demonstrable in these mice but not in STZ-treated mice. The inability of STZ-treated tumor-bearing mice to elicit a delayed type hypersensitivity response was not due to selective depletion of delayed type hypersensitivity-inducing CD4+ T-cells but was probably due to failure of STZ to eliminate tumor-specific suppressor cells. Together these findings suggested that the failure of STZ to cure LSA tumor-bearing mice was not due to lack of tumoricidal activity or related to suppression of tumor-specific effector T-cell function but may be due to the failure of STZ to eliminate tumor-specific T suppressor cells. The present study suggests that the outcome of chemotherapy with nitrosoureas depends, in addition to the tumoricidal activity of the drug, on the immunomodulating action on the immune mechanisms of the host. PMID- 2573419 TI - Amplification, overexpression, and rearrangement of the erbB-2 protooncogene in primary human stomach carcinomas. AB - Four of 51 primary gastric carcinomas exhibited amplification of the erbB-2 protooncogene ranging from 2- to 8-fold. In three cases gene amplification affected erbB-2 alleles of normal gene structure as determined by Southern blot analysis. In addition, one tumor displayed gene amplification of an apparently rearranged erbB-2 allele. Analysis of the rearranged allele revealed a structural alteration consistent with an internal deletion of approximately 2 kilobase pairs within the erbB-2 gene. Quantitation of erbB-2 mRNA in these tumors demonstrated that gene amplification coincided with overexpression of erbB-2 mRNA ranging from 8- to 32-fold above levels observed in stomach tumors without gene amplification. Furthermore, in one of the tumors with amplified normal size erbB-2 restriction fragments, elevated erbB-2 mRNA levels consisted of the normal size 5-kilobase transcript. Thus, overexpression of normal size erbB-2 mRNA accompanies gene amplification in primary stomach tumors. In addition, evidence for erbB-2 gene rearrangement suggests a role for such alteration in the development of certain gastric carcinomas. PMID- 2573420 TI - Genetic alterations of c-myc, c-erbB-2, and c-Ha-ras protooncogenes and clinical associations in human breast carcinomas. AB - We have analyzed genomic DNA sequences from 125 prospectively collected single unilateral primary breast carcinoma samples for the presence of alterations of c myc, c-erbB-1, c-erbB-2, c-Ki-ras and c-Ha-ras protooncogenes. Amplification of the c-myc gene was found in 18% of the samples, and in one sample a non-germ line c-myc related DNA fragment or rearrangement was detected. We have found a significant association (P = 0.0010) between amplified c-myc gene and inflammatory carcinoma, a particularly aggressive breast cancer. The c-erbB-2 gene was amplified in 22% of the tumor samples and a rearrangement was observed once. Alteration of the c-erbB-2 gene was significantly linked to histological grade III tumors (P = 0.005) and the absence of estrogen and progesterone receptors (P = 0.036). No amplifications were observed for c-erbB-1, c-Ki-ras, and c-Ha-ras genes. About 40% of breast carcinomas contain either amplified c-myc or c-erbB-2 protooncogenes, whereas simultaneous amplification of both was seen in only one sample, suggesting the involvement of two distinct molecular mechanisms in breast cancer. Comparison of DNA from peripheral blood and tumor samples indicated loss of one c-Ha-ras allele in 29% of patients heterozygous for this polymorphism. A significant correlation (P = 0.016) between c-Ha-ras locus (11p14) allele loss and patient survival was found. These data suggest that 11p14 allelic loss plays a role in the evolution of human breast cancer, amplification of c-erbB-2 gene is associated with increasing stage of malignancy, and alteration of the c-myc gene in inflammatory breast carcinoma may contribute to the rapid progression of this human tumor subtype. PMID- 2573421 TI - Molecular aspects of growth control. An extended report on the first joint meeting of the AACR and the Japanese Cancer Association. PMID- 2573422 TI - Increased expression of sialyl-dimeric LeX antigen in liver metastases of human colorectal carcinoma. AB - We collected a total of 78 tissue specimens, including primary colorectal carcinoma, normal colonic mucosa, and liver metastases of colon carcinoma, to examine whether the extracts of these tissues inhibited the binding of a monoclonal antibody FH6, specific for sialyl-dimeric LeX antigen. The results of inhibition assays demonstrated that: (a) contents of FH6-reactive molecules were greater in carcinoma tissues than in normal colonic mucosa; (b) metastatic foci in livers contained more FH6-reactive molecules than primary tumors; (c) primary tumors from Dukes' stage B1 patients contained less FH6-reactive molecules than primary tumors from Dukes' stage D patients. The inhibitory activity of these tumor tissue extracts against the binding of a monoclonal antibody FH6 to cultured colon carcinoma cells was eliminated by prior treatment of the extracts with sialidase, confirming that the FH6-reactive materials were sialyl-dimeric LeX antigen. Electrophoretic separation of tumor tissue extracts on 3% polyacrylamide gels followed by direct staining with monoclonal antibody FH6 revealed that very high molecular weight glycoproteins, presumably mucins, contained sialyl-dimeric LeX antigen. PMID- 2573424 TI - c-erbB-2 amplification in node-negative human breast cancer. AB - c-erbB-2 gene analysis by Southern and DNA dot blot methods was done in 66 tumor samples from patients with histologically node-negative breast cancer. The c-erbB 2 gene was amplified 2- to greater than 8-fold in 13 tumors (20%). None of 59 tumors that were examined by the Southern method showed c-erbB-2 gene rearrangement. c-erbB-2 amplification was analyzed in relation to other prognostic factors. The c-erbB-2 gene was amplified in five of 36 (14%) diploid and eight of 30 (27%) aneuploid tumors. Thirteen of 54 (24%) tumors with nuclear Grade 1 or 2 displayed c-erbB-2 amplification, whereas none of 12 tumors with nuclear Grade 3 did. No correlation was observed with estrogen receptor content, tumor size, histological type, or age of patients. The median follow-up date for these patients was 85+ mo. Of 13 patients whose tumors showed c-erbB-2 amplification, six patients (46%) developed recurrence, and five patients (38%) died of metastatic disease. In contrast, of 53 patients whose tumors did not show c-erbB-2 amplification, 15 patients (28%) developed recurrence, and seven patients (13%) died of disease. In conclusion, our results show that c-erbB-2 gene amplification was more frequent in aneuploid tumors and tumors with poor nuclear grade. c-erbB-2 amplification may be considered a possible prognostic factor in node-negative breast cancer. PMID- 2573423 TI - Mechanisms of inhibition of DNA synthesis by 2-chlorodeoxyadenosine in human lymphoblastic cells. AB - 2-Chloro-2'-deoxyadenosine 5'-triphosphate (CldATP) is a strong inhibitor of the reduction of ADP, CDP, UDP and GDP by ribonucleotide reductase in extracts of CCRF-CEM with 50% inhibition at concentrations of 0.1 to 0.3 microM. In cells exposed to 0.3 microM 2-chloro-2'-deoxyadenosine (CldAdo), the intracellular concentration of CldATP reaches 2 microM within 15 min, and DNA synthesis by the cells is inhibited 90% within 30 min. At concentrations of extracellular CldAdo that inhibit DNA synthesis, there is also marked inhibition of intracellular conversion of cytidine to deoxycytidine nucleotides indicating significant intracellular inhibition of ribonucleotide reductase. Exposure of cells to 0.3 microM CldAdo decreases dCTP by 63% in 30 min, dATP and dTTP by 20% and dGTP by a smaller amount. Similar decreases in these pools occur when other inhibitors of ribonucleotide reductase are present at concentrations causing similar inhibition of DNA synthesis. Deoxycytidine treatment of cells inhibited by CldAdo restores dCTP and other pools, but restoration of DNA synthesis is incomplete, indicating that there is another mechanism for inhibition of DNA synthesis in addition to depletion of deoxyribonucleotide pools. This alternate mechanism is probably related to the incorporation of CldAdo into DNA that occurs despite a 25-times lower intracellular level of CldATP than dATP. PMID- 2573425 TI - Systemic induction of cells mediating antibody-dependent cellular cytotoxicity following administration of interleukin 2. AB - We have previously demonstrated that incubation of murine cells in vitro in interleukin 2 (IL-2) induced antibody-dependent cellular cytotoxicity (ADCC) and that these cells were derived from the NK/LAK, FcR+ cell population. In the present study we show that in vivo administration of IL-2 to mice induces cells which exhibit ADCC activity in the peritoneal cavity, liver, lungs, and to a lesser degree in the bone marrow, spleen, mesenteric lymph nodes, and thymus. A gradual increase in ADCC activity and the number of Fc-receptor-positive cells was seen 1 to 3 days after starting IL-2 treatment. The cells mediating ADCC are closely related to LAK cells since they expressed Thy1.2 antigens and are derived from asialo GM1-positive, Lyt2/L3T4-negative, radiosensitive cells. These results demonstrate that IL-2 can systemically induce cells with ADCC activity and that this ability may be useful in the establishment of therapeutic models against disseminated cancer when combined with specific antitumor monoclonal antibodies. PMID- 2573427 TI - [An alpha-satellite DNA sequence, alpha-RI-6, specific for human chromosomes 13 and 21, detected using the RFLP technic with digoxigenin labelled probes]. AB - The authors compared two at present most widely used techniques for labelling DNA probes: a) radioactive labelling by means of the radioisotope 32P; non radioactive labelling using the hapten digoxigenin for the visualization of the hybridization process on nylon membranes. Then sensitivity of the technique of non-radioactive labelling of heterochromatin probes was equivalent to the radioactive method. PMID- 2573426 TI - Analysis of c-erbB-2 expression in breast carcinomas with clinical follow-up. AB - Various monoclonal antibodies reactive with protooncogene products or tumor associated antigens have been utilized to investigate breast carcinoma biology or antigen expression with potential prognostic relevance. Murine monoclonal antibody TA1, generated by immunization of BALB/c mice with whole c-erbB-2 (neu) transformed NIH/3T3 cells, recognizes the extracellular domain of the c-erbB-2 protein and binds a Mr 185,000 protein by immunoprecipitation. Using avidin biotin-peroxidase techniques and monoclonal antibody TA1, 313 archival primary adenocarcinomas of the breast were evaluated for c-erbB-2 overexpression; 290 of these were used for multiparametric statistical analysis. Historical, clinical (age, laterality), histological (nuclear grade, tumor size, lymph node status, lymphatic or blood invasion), and hormone receptor data as well as clinical outcome (minimal follow-up, 6 years; median follow-up, 8.5 years) were compared to TA1 staining. For these 290 patients Cox regression multivariate analysis showed the strongest correlation between lymph node status or estrogen receptor status and overall survival (P = 0.0001 and 0.049, respectively). TA1 staining did not significantly correlate with survival (P = 0.395). However, univariate analysis of certain patient subpopulations showed a significant correlation if the examined tumors were subdivided into negative or focally reactive and those with greater than or equal to 40% cellular reactivity. For T3, T4 patients, strong TA1 immunoreactivity correlated with a shortened disease-free survival (log rank P = 0.0018; Wilcoxon p = 0.0078) and overall survival (log rank P = 0.0002; Wilcoxon P = 0.0013). For these patients the overall survival at 6 years was markedly different between the strongly reactive tumors (0%) and the negative to weakly reactive tumors (55%). In lymph node-positive patients a trend between high TA1 reactivity and a worse overall survival was also noted (log rank P = 0.128; Wilcoxon P = 0.054), with a 6-year survival of 42% in the strongly reactive tumors (n = 16) and 65% in the negative to weakly reactive carcinomas (n = 105). No correlation between TA1 immunoreactivity and other historical, clinical, and histological features were noted. c-erbB-2 overexpression as measured by immunohistochemical techniques, therefore, may have clinical significance in certain patient subpopulations. PMID- 2573428 TI - Binding and internalization of gold-conjugated somatostatin and growth hormone releasing hormone in cultured rat somatotropes. AB - The synthetic peptides somatostatin (SRIF) and growth hormone-releasing hormone (GRH) were coupled directly to colloidal gold of different particle sizes. Both conjugates were biologically active in displacing the corresponding radiolabeled hormones from high affinity binding sites in pituitary membranes. Release of growth hormone (GH) from cultured anterior pituitary cells was modulated by both conjugates alone or in combination. Ultrastructural studies were performed with cells incubated at 4 degrees C (2 h) and 37 degrees C (2 min-2 h) with one of the labeled peptides or their combination. Somatotropes were identified by immunostaining with anti-rGH followed by protein A-ferritin, thus obtaining a triple labeling. Both hormone conjugates were internalized in different vesicles in the beginning but accumulated during longer incubation times in the same compartment. The secretory vesicles and the nucleus were not labeled by any hormone conjugate. In contrast to SRIF-gold, the uptake of GRH-gold conjugate decreased with longer incubation times. This effect could be neutralized by simultaneous incubation of the somatotropes with both regulating hormones. Hence, whereas the binding and internalization of SRIF by somatotropes do not seem to be influenced by GRH, the corresponding processes for GRH are stimulated by the presence of SRIF. PMID- 2573429 TI - Adrenergic neurons and short proprioceptive feedback loops involved in the integration of cardiac function in the rat. AB - Serial cryostat and paraffin-embedded sections through the atrioventricular junction of the rat heart were studied at the light-microscopic level after indirect immunohistochemical staining (tyrosine hydroxylase, neuropeptide Y, C terminal flanking peptide of neuropeptide Y immunoreactivities) or silver impregnation. The distribution of these immunoreactivities in the Hissian ganglion (Moravec and Moravec 1984) as well as the relationships of the Hissian ganglion cells with the surrounding structures have been studied to assess its function. The results suggest that the Hissian ganglion is composed of large multipolar neurons displaying both tyrosine hydroxylase (TH) and related peptide (neuropeptide Y. C-terminal flanking peptide of neuropeptide Y) immunoreactivities. The dendritic projections of these adrenergic cells penetrate the reticular portion of the atrioventricular node and the upper segments of the interventricular septum where they constitute sensory-like corpuscles. The hypothesis that the adrenergic neurons of the atrioventricular junction are involved in short proprioceptive feedback loops necessary for beat-to-beat modulation of cardiac excitability and intracardiac conduction can thus be suggested. PMID- 2573430 TI - Polypeptide chain binding proteins: catalysts of protein folding and related processes in cells. AB - Subcellular compartments in which folding and assembly of proteins occur seem to have a set of PCB proteins capable of mediating these and related processes, such as translocation across membranes. When a domain of a polypeptide chain emerges from a ribosome during synthesis or from the distal side of a membrane during translocation, successive segments of the chain are incrementally exposed to solvent and yet are unlikely to be able to fold. This topological restriction on folding likely mandates a need for PCB proteins to prevent aggregation. Catalysis of topologically restricted folding by PCB proteins is likely to involve both an antifolding activity that postpones folding until entire domains are available and, more speculatively, a folding activity resulting from a programmed stepwise release that employs the energy of ATP hydrolysis to ensure a favorable pathway. We are left with a new set of problems. How do proteins fold in cells? What are the sequences or structural signals that dictate folding pathways? The new challenge will be to understand folding as a combination of physical chemistry, enzymology, and cell biology. PMID- 2573431 TI - Cyclic AMP stimulates somatostatin gene transcription by phosphorylation of CREB at serine 133. AB - In this paper, we demonstrate that phosphorylation of CREB at Ser-133 is induced 6-fold in vivo, following treatment of PC12 cells with forskolin. By contrast, no such induction was observed in the kinase A-deficient PC12 line A126-1B2 (A126). Using F9 teratocarcinoma cells, which are unresponsive to cAMP, we initiated a series of transient expression experiments to establish a causal link between phosphorylation of CREB and trans-activation of cAMP-responsive genes. Inactivating the kinase A phosphorylation site by in vitro mutagenesis of the cloned CREB cDNA at Ser-133 completely abolished CREB transcriptional activity. As CREB mutants containing acidic residues in place of the Ser-133 phosphoacceptor were also transcriptionally inactive, these results suggest that phosphorylation of CREB may stimulate transcription by a mechanism other than by simply providing negative charge. PMID- 2573432 TI - Precursor frequency of human T4 cells responding to stimulation through the CD3 molecular complex: role of various cytokines in promoting growth and IL2 production. AB - The frequency of human T4 cells induced to grow and produce IL2 in response to the anti-CD3 mAb, 64.1, was examined. T4 cells were cultured at limiting dilution and stimulated with either soluble or immobilized 64.1 in the presence of various cytokines and/or irradiated B lymphoblastoid cells as accessory cells (AC). The frequency of responding cells was assessed by examining wells microscopically for visible growth and supernatants for IL2. Immobilized, but not soluble, 64.1 was able to induce T4 cells to grow in the complete absence of AC, but only when exogenous cytokines were present. IL2 was most effective at supporting T4 cell growth in this system, with a mean of 26.0 +/- 3.8% of immobilized 64.1-activated T4 cells generating a colony in cultures supplemented with IL2. IL4 could also support the growth of immobilized 64.1-activated T4 cells, but the frequency of responding cells was much lower (3.7 +/- 0.9%). The combination of IL2 and IL4 was not more effective than IL2 alone. TNF alpha, IL1 beta, and IL6 were unable to support T4 cell growth alone, but each increased the frequency of T4 cells responding in the presence of IL2. AC could support the growth of a small number of 64.1-stimulated T4 cells in the absence of exogenous IL2 and enhanced the frequency of T4 cells responding to immobilized 64.1 in the presence of IL2. The percentage of immobilized 64.1-stimulated T4 cells producing IL2 was also examined. Immobilized 64.1 stimulated less than 1.4 in 1000 T4 cells to produce IL2 in the absence of AC and neither IL4 nor TNF alpha enhanced this response. Fixed AC and IL1 beta, on the other hand, caused a small increase in the frequency of immobilized 64.1-activated T4 cells that secreted IL2. The frequency of T4 cells stimulated to produce IL2 by immobilized 64.1 was greatly enhanced by the addition of AC. The data indicate that in the absence of AC, a stimulatory matrix of immobilized 64.1 is sufficient for some T4 cells to be activated to become IL2 or IL4 responsive and for a smaller percentage to secrete IL2. Additional T4 cells require IL1 beta, TNF alpha, IL6, or AC to become IL2 responsive, whereas only IL1 beta and AC can promote IL2 production. In the presence of AC, the amount of cytokine produced endogenously appears to be sufficient to sustain the growth of some T4 cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2573433 TI - The myelopoietic inducing potential of mouse thymic stromal cells. AB - The thymus has generally been considered as being solely involved in T cell maturation. In this study we have demonstrated that mouse thymic stroma can also support myelopoiesis. Bone marrow from mice treated with 5-fluorouracil was depleted of cells expressing Mac-1, CD4, and CD8 and incubated on lymphocyte-free monolayer cultures of adherent thymic stromal cells. After 7 days there was a marked increase in nonadherent cells, the majority of which were Mac-1+, FcR+, and HSA+. These proliferating bone marrow cells also expressed markers (MTS 17 and MTS 37) found on thymic stromal cells. Such cells were not found in thymic cultures alone, in bone marrow cultured alone, or on control adherent cell monolayers. Supernatants from the cultured thymic stroma, however, were able to induce these cell types in the bone marrow precursor population. Incubation of normal thymocytes with a monolayer of these in vitro cultivated Mac-1+, MTS 17+, MTS 37+ myeloid cells leads to selective phagocytosis of CD4+ CD8+ cells. Hence, this study demonstrates that the thymic adherent cells can induce myelopoiesis in bone marrow-derived precursor cells and provide a form of self-renewal for at least one population of thymic stromal cells. Furthermore, these induced cells are capable of selective phagocytosis of CD4+ CD8+ thymocytes and may provide one mechanism for the selective removal of such cells from the thymus. PMID- 2573434 TI - T lymphocytes and their CD4 subset are direct targets for the inhibitory effect of calcitriol. AB - We studied the direct effects of the hormone calcitriol on the activation and proliferation of pure T lymphocytes and their subsets. Calcitriol inhibited the proliferation of T lymphocytes stimulated in the absence of monocytes with phytohemagglutinin (PHA) and either a monocytic culture supernatant or a combination of monocyte-derived interleukin 1 and interleukin 6. This inhibition was not influenced by the concentration of the stimulating agents. The minimal effective concentration of calcitriol was 10(-10) M. In contrast, the interleukin 2 (10 U/ml)-driven growth of PHA-stimulated T lymphocytes was not significantly altered by calcitriol at 10(-8) M. The hormone had also no influence on the T lymphocyte proliferation induced by a combination of PHA and the anti-CD28 monoclonal antibody 9.3. Pure T lymphocytes, after incubation for 5 days with PHA and monocytic factors, expressed a high level of transferrin receptors. This phenomenon was strongly suppressed on both CD4 and CD8 subsets when 10(-8) M calcitriol had been present during the culture. Moreover, the proliferation of pure CD4 cells was directly inhibited by calcitriol in similar conditions as for unseparated T lymphocytes. We conclude that T lymphocytes and their CD4 subset are direct targets for the inhibitory effect of calcitriol. PMID- 2573435 TI - Activation requirements of donor T cells and host T cell recruitment in adoptive transfer of murine experimental autoimmune orchitis (EAO). AB - The relative roles of donor and host T lymphocytes and the T cell activation requirements in adoptive transfer of experimental autoimmune orchitis (EAO) in (C57BL/6 x A/J)F1 mice were investigated in order to gain an understanding of the pathogenesis of this disease. Depletion of T cell subsets in recipients by adult thymectomy and treatment with monoclonal antibodies against CD4 or CD8 had no effect on the incidence of EAO following adoptive transfer of activated T cells from donors immunized with testis homogenate (TH) and adjuvants. In contrast, such depletion of CD4+ T cells inhibited development of EAO in actively immunized mice. Thus, CD4+ cells are required for induction of EAO, but donor CD4+ cells are sufficient by themselves without a comparable contribution from the recipient. Adoptive transfer of EAO required that donor splenic and lymph node T cells be activated in vitro before transfer. We found that exposure to antigen (TH) for as little as 4 hr allowed EAO to occur in 25% of recipients, and by 24 hr the cells were fully competent to induce disease. Proliferation of the cells could not be measured until 2 days later. In serial double-transfer experiments, it was found that the cells must be cultured with TH before each transfer in order for the secondary recipients to develop EAO. However, it was not necessary for the transferred T cells to "see" antigen in vivo in the primary recipients, since transfer to castrated primary recipients had no effect on EAO incidence in secondary recipients. Lymphocytes isolated from diseased testes of immunized donors were competent to transfer EAO without activation in vitro, suggesting that, unlike spleen and lymph node cells, these orchitic lymphocytes were already capable of trafficking to the testis. PMID- 2573436 TI - Neuromuscular and cardiovascular effects of mivacurium chloride in surgical patients receiving nitrous oxide-narcotic or nitrous oxide-isoflurane anaesthesia. AB - The neuromuscular and cardiovascular effects of mivacurium chloride were studied during nitrous oxide-oxygen narcotic (fentanyl) (n = 90) and nitrous oxide-oxygen isoflurane (ISO) anaesthesia (n = 45). In addition, a separate group (n = 9) received succinylcholine during fentanyl anaesthesia to compare its neuromuscular effects with mivacurium. Mivacurium was initially administered as a single bolus in doses from 0.03 mg.kg-1 to 0.25 mg.kg-1 to study the dose-response relationships, as well as the cardiovascular effects of mivacurium. Neuromuscular block (NMB) was measured by recording the twitch response of the adductor pollicis muscle following ulnar nerve stimulation (0.15 Hz, 0.2 ms supramaximal voltage). The ED95 values for mivacurium were estimated to be 0.073 mg.kg-1 and 0.053 mg.kg-1 in the fentanyl and ISO groups respectively. The duration of block (time from injection to 95 per cent recovery) for a dose of 0.05 mg.kg-1 mivacurium was 15.3 +/- 1.0 min and 21.5 +/- 1.3 min for fentanyl and ISO anaesthesia, respectively. The recovery index (25-75 per cent) between initial bolus dose (6.1 +/- 0.5 min), repeat bolus doses (7.6 +/- 0.6 min), mivacurium infusion (6.7 +/- 0.7 min) and succinylcholine infusion (6.8 +/- 1.8 min) were not significantly different. There was minimal change in mean arterial pressure (MAP) or heart rate (HR) following bolus doses of mivacurium up to 0.15 mg.kg-1. Bolus administration of 0.20 mg.kg-1 or 0.25 mg.kg-1 of mivacurium decreased MAP from 78.2 +/- 2.5 to 64.0 +/- 3.2 mmHg (range 12-59 per cent of control) (P less than 0.05). The same doses when administered slowly over 30 sec produced minimal change in MAP or HR. PMID- 2573437 TI - Contribution of the baroreflex afferent nerves to the production of vasoconstricted hypertension in volume-expanded dogs. AB - Dextran in lactated Ringer's solution (20 ml/kg) was infused for 1 hour into anesthetized dogs with sinoaortic denervation and vagotomy (deafferentation; n = 10) and dogs treated with hexamethonium (de-efferentation; n = 13) to compare with our previous observation in dogs with an intact autonomic nervous system (control, n = 34). During the infusion, increase in blood pressure associated with increase in cardiac output was observed in all three groups. The increases in blood pressure were larger in the two groups with an impaired autonomic nervous system. In the recovery period, the control dogs and the hexamethonium treated dogs showed gradual increases in total peripheral resistance and in vasoconstricted hypertension 3 hours after stopping the infusion. In contrast, the dogs with sinoaortic denervation and vagotomy did not show any increase in total peripheral resistance. The vasoconstricted groups showed peaks of natriuresis soon after the infusion, not 3 hours after the infusion when vasoconstriction was observed, although the dogs with deafferentation did not show a significant increase in natriuresis. Norepinephrine (0.5 micrograms/kg) was administered intravenously before and after volume expansion, and the pressor responses in the three groups after volume expansion were enhanced similarly (143%, 128%, and 136%, respectively). These results indicate that the afferent signals from peripheral vessels to the brain contribute to the production of vasoconstricted hypertension after acute volume expansion and that the vasoconstriction is independent of pressor hypersensitivity and is dissociated in time from the natriuresis. PMID- 2573438 TI - Canine neutrophil activation by cardiac lymph obtained during reperfusion of ischemic myocardium. AB - Cardiac lymph from a canine model of myocardial ischemia and reperfusion was examined for evidence of chemotactic activity. Lymph was continuously collected from awake animals before and during a 60-minute coronary artery occlusion and up to 6 hours after the initiation of reperfusion. It was assessed for the ability to activate the following proinflammatory functions in neutrophils isolated from the blood of healthy dogs: 1) morphological changes characteristic of chemotactic stimulation, which were assessed by phase contrast microscopy, 2) orientation of canine neutrophils in a gradient of cardiac lymph, which was assessed in Zigmond chambers, 3) the binding of monoclonal antibodies reactive with CD11b and CD18 adherence glycoproteins, which was assessed by flow cytometry, and 4) adherence of canine neutrophils to monolayers of canine jugular vein endothelium, which was assessed in vitro by a visual assay. Lymph samples collected after 1 hour of reperfusion in animals demonstrating ECG evidence of ischemia and histological evidence of infarction exhibited significant stimulatory activity for each of the functions tested. Shape change-inducing activity was evaluated at more frequent intervals than other functions and was found to peak at 1 hour after initiation of reperfusion and to disappear by 6 hours. In addition, the CD11b/CD18 levels on neutrophils isolated from cardiac lymph collected during reperfusion were significantly greater than neutrophils obtained before or during occlusion. Animals that failed to exhibit evidence of infarction also failed to exhibit increased stimulatory activity in lymph collected during reperfusion, and surface levels of CD11b/CD18 on neutrophils collected from reperfusion lymph were not elevated. This study provides direct evidence supporting the hypothesis that chemotactic activity is generated in ischemic and reperfused myocardium. PMID- 2573440 TI - T. Lauder Brunton, 1844-1916. PMID- 2573439 TI - In vitro electrophysiological analysis of mature rat hippocampal transplants in oculo. AB - We have investigated the maturation of isolated rat hippocampus grafted into the anterior chamber of the eye. Electrophysiological responses from transplants were compared to those recorded from the in vitro hippocampal slice preparation. Intracellular recording demonstrated that the passive membrane characteristics of intraocular hippocampal neurons were similar to those of the CA1 pyramidal cells in the in vitro slice preparation. However, the slow after-hyperpolarization which normally follows depolarization-induced action potentials was reduced or completely absent in the intraocular transplants, and the excitatory postsynaptic potential (EPSP) evoked by local stimulation was prolonged. The duration of the EPSP was reduced by perfusion with D-aminophosphonovaleric acid (2.5-50 microM), an N-methyl-D-aspartate receptor antagonist. Normal levels of glutamate decarboxylase (a marker for gamma-aminobutyric acidergic neurons) were found in the transplants, and responses to adenosine, bicuculline, and norepinephrine were similar in the in oculo transplants and in vitro slices. The data suggest that although many properties of hippocampal neurons are intrinsically determined, other aspects of the physiology of mature hippocampus either fail to develop, or develop abnormally in the absence of external inputs in oculo. PMID- 2573441 TI - Paired stimulation and functional change of the sural nerve during beta blocker therapy. AB - 29 patients were examined during a therapy with beta blockers. The medication was applied over 12 weeks, 16 patients received propranolol 120 mg/day, 13 patients metoprolol 200 mg/day. Two patients showed side-effects. Neurophysiological examinations, i.e. sural nerve conduction velocity and paired stimulation of the sural nerve, revealed a slight decrease of nerve conduction velocity and a significant increase of the latency prolongation of the second nerve action potential when the medication was finished. These functional changes in the sural nerve may result from the interaction of metoprolol and propranolol with beta receptors of the peripheral nerve. PMID- 2573443 TI - Indenolol, a beta-blocker with partial agonism at vascular beta-adrenoceptors. AB - Intrinsic sympathomimetic activity may attenuate some effects caused by treatment with beta-adrenoceptor blockers. Indenolol is a nonselective beta-adrenoceptor antagonist whose sympathomimetic properties have been shown in vitro but not in human beings. We infused indenolol cumulatively (5, 15, and 50 micrograms/100 ml tissue per minute for 15 minutes each, preceded by an infusion of saline solution) into the brachial arteries of nine hypertensive patients. Forearm blood flow (venous plethysmography), mean arterial pressure, and heart rate were monitored. During infusion at 5 micrograms/100 ml tissue per minute, forearm blood flow did not change, but it did increase dose-dependently at the greater infusion rates. This action was determined to be mediated by beta-adrenoceptor stimulation because propranolol (10 micrograms/100 ml tissue per minute for 15 minutes), given before treatment and then concomitantly with indenolol, abolished it (n = 5). Indenolol vasodilated forearm arterioles and this effect was antagonized by beta-blockade, thus demonstrating vascular intrinsic sympathomimetic activity. This property may contribute to its therapeutic action in human beings. PMID- 2573444 TI - Clinical evaluation of a benzofuroquinolizine alpha 2-adrenoceptor antagonist. AB - The pharmacodynamics of MK-912, a benzofuroquinolizine alpha-adrenoceptor antagonist, were evaluated in healthy male volunteers. Eight subjects were treated with single oral doses of 0.1, 1.0, and 2.0 mg MK-912 and with a placebo in a four-period, double-blind, balanced, crossover study. Hemodynamic effects were observed with the 2.0 mg dose of MK-912 (peak increase from baseline in systolic and diastolic blood pressure +/- SEM, 14.8/9.2 +/- 2.9/2.1 mm Hg; peak increase in heart rate, 6.3 +/- 2.1 beats/min; p less than 0.05 versus placebo). Plasma concentrations of 3-methoxy-4-hydroxyphenylglycol (MHPG, a catecholamine metabolite) were increased 29% +/- 7% and 40% +/- 10% above baseline 2 hours after administration of 1.0 and 2.0 mg MK-912, respectively (p less than 0.01 compared with placebo). A modest dose-dependent reduction (5% to 10%) in fasting plasma glucose concentration was observed 1/2 to 1 hour after administration of 1.0 and 2.0 mg MK-912 (p less than 0.05 compared with placebo), without significant change in plasma insulin values. MK-912 was well tolerated, although it did have a mild anxiogenic effect. MK-912 is a potent, orally active agent with a pharmacologic profile consistent with alpha 2-adrenoceptor antagonism. PMID- 2573445 TI - Predictability of antihypertensive responsiveness and alpha-adrenoceptor antagonism during prazosin treatment. AB - This study investigates concentration-effect relationships in nine patients who had essential hypertension treated with prazosin. Antihypertensive responsiveness was determined for each individual patient in terms of millimeters of mercury of blood pressure reduction per nanogram per milliliter of plasma prazosin concentration. The principal findings were that there was significant attenuation of antihypertensive responsiveness, from 11.5 mm Hg per nanogram per milliter after the first dose to 8.7 mm Hg per ng/ml after 1 week of treatment with prazosin. Correspondingly, there was significant attenuation of the degree of alpha 1-antagonism as assessed by the pressor response to intravenous phenylephrine. However, there was no significant further attenuation of either assessment during continued treatment for up to 3 months. These findings suggest that after an early adaptation, which occurs within the first week of treatment, there is no long-term attenuation of the antihypertensive effect of prazosin. Despite this initial adaptive change, the magnitude of the long-term antihypertensive effect of prazosin was predictable from the first dose response in individual patients. PMID- 2573442 TI - Drug interactions involving aspirin (acetylsalicylic acid) and salicylic acid. AB - Aspirin (acetylsalicylic acid) is metabolically converted to salicyclic acid by the action of carboxylesterases. Although metabolic drug interactions involving aspirin are theoretically possible, there appear to have been no studies to date which have shown conclusively that aspirin hydrolysis is altered by coadministered drugs. However, a number of treatments are known to affect the rate or extent of aspirin absorption, including activated charcoal, antacids, cholestyramine and metoclopramide. Caffeine and metoprolol have been reported to increase peak salicylic acid concentration following aspirin administration, and coadministration of dipyridamole and aspirin results in higher plasma aspirin concentrations. The mechanism(s) responsible for these latter observations remains unknown. Salicylic acid is extensively bound to plasma albumin, and many of the reported drug interactions involve displacement of the coadministered drug from plasma protein. Protein binding displacement appears to be the basis of salicylic acid interactions with diclofenac, flurbiprofen, ibuprofen, isoxicam, ketoprofen, naproxen, phenytoin and tolmetin. Following displacement of these agents increased clearance of total drug occurs, and consequently the plasma concentration of total drug decreases. Although generally not measured, unbound concentration of the interacting drug should not be markedly altered. Salicylic acid also increases total plasma clearance of fenoprofen but, unlike the interactions with the other propionic acid non-steroidals, plasma protein binding displacement does not appear to be involved. Induction of fenoprofen metabolism is a possibility, although there is no firm evidence from other studies that salicylate is able to induce the metabolism of coadministered drugs. Since salicylic acid is extensively metabolised, it is not surprising that it is able to inhibit the metabolism of certain coadministered drugs and chemicals, an effect which has been reported for salicylamide, valproic acid, m-xylene, and zomepirac. The interactions with salicylamide, m-xylene and zomepirac are probably competitive in nature since mutual inhibition of salicylic acid metabolism occurs. There is an additional component of protein binding displacement in the interactions with valproic acid and zomepirac, resulting in increased unbound drug concentration. Certain coadministered drugs (or chemicals) may alter the metabolism of salicylic acid; inhibition of its metabolism has been demonstrated following treatment with benzoic acid, salicylamide, m-xylene, zomepirac and possibly cimetidine. In contrast, salicylic acid elimination is enhanced in oral contraceptive steroid users and by corticosteroid treatment. Oral contraceptive steroids induce both salicylic acid glucuronidation and salicylurate formation. Induction of metabolism has also been proposed to account for the effects of corticosteroids, but this is still to be proven.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2573446 TI - Lack of an association between diffuse systemic sclerosis and HLA-DR1 or HLA-DR5. AB - The HLA-DR locus of the major histocompatibility complex encodes class II molecules which participate in immune responses through regulation of T cell interaction with antigen presenting cells. Previous association studies between HLA-DR antigens and the autoimmune disease, systemic sclerosis (or scleroderma), have yielded conflicting results. Some investigators have reported an association between this disease and HLA-DR1, while others have demonstrated an association with HLA-DR5. In this study, we used restriction fragment length polymorphisms in the HLA-DR locus to compare allelic frequencies of HLA-DR1 and HLA-DR5 in scleroderma patients with diffuse disease and healthy control subjects. No significant difference in the allelic frequency of either antigen was observed between the groups. These results suggest that HLA-DR1 and HLA-DR5 antigens are unlikely to contribute significantly to disease susceptibility in scleroderma. PMID- 2573447 TI - Application of synthetic oligonucleotides to detect DQ beta genes transmission within insulin-dependent diabetes families. AB - Class II antigen genes encoded by the major histocompatibility complex region (HLA-D region) in man play an important role in susceptibility to insulin dependent diabetes mellitus (IDDM). Evidence suggests that the DQ subregion within the HLA-D region is more directly responsible for susceptibility to IDDM. Therefore, we designed a synthetic oligonucleotide specific for the DQ beta gene to further the understanding of the disease association with HLA-D region genes at the molecular level. Restriction fragment length polymorphism (RFLP) analysis was carried out using DNA isolated from nine families, each including at least two affected siblings (a total of 37 siblings). The segregation pattern of hybridizing fragments showed that: (1) for each of the DR2, DR3, and DR4 specificities, two different alleles can be identified by the DQ beta probe; (2) a 1.9 kb-Taq 1 fragment with the DR4 specificity and a 6.0 kb-Taq-1 fragment within the DR2 specificity tend to cosegregate with IDDM; (3) there was no preferential segregation of the two alleles detected within the DR3 specificity (one allele identified by a 4.7 kb-Taq 1 fragment is quite common among individuals with the DR3 specificity). The results from this study add to the evidence that certain DQ alleles appear to be more directly associated with the diabetogenic gene (or genes) in certain DR specificities. PMID- 2573448 TI - Assignment of an anonymous DNA sequence IB/D22 (DXS371) to XP11.1-11.4. AB - In densely mapped and characterized chromosomes such as the X chromosome there are still regions which have a paucity of markers. One of these areas lies in the Xp11 region. Using human-rodent somatic cell hybrids we have assigned a newly isolated clone IB/D22 to the Xp11.1-p11.4 region. PMID- 2573449 TI - Comparative physiology and biochemistry: challenges for the future. AB - 1. Comparative physiology is distinguished from other types of physiology by treating the diversity of solutions of functional problems and by using kind of animal as a functional variable. 2. The strength of comparative physiology os its capacity to give some solutions to problems in basic biology. 3. Specific examples of subject areas to which comparative physiology contributes are: (a) mechanisms underlying evolution; (b) the nature of speciation; (c) comparative cognitive science, neural models for behavior; and (d) applications of molecular techniques to physiology of whole animals. 4. Applications continue in ecology, medicine and agriculture. 5. The breadth of the comparative approach to physiology has important philosophical implications. PMID- 2573450 TI - Respiration and osmoregulation of the estuarine crab, Rhithropanopeus harrisii (Gould): effects of the herbicide, alachlor. AB - 1. The effects of a sudden decrease in salinity and exposure to sublethal concentrations of the herbicide, alachlor, on osmoregulation and respiration of the crab, Rithropanopeus harrisii, were studied. 2. Crabs were hyperosmotic regulators at salinities below 24 ppt and became hypoosmotic at higher salinities. Upon a salinity decrease from 20 to 1 ppt, crabs adjusted their haemolymph osmolality to a stable hyperosmotic level in 8 hr. Alachlor concentrations to 50 ppm did not affect this adjustment. 3. A salinity decrease from 10 to 0 ppt elevated VO2 and the critical oxygen tension. This response was unaffected by alachlor concentrations as high as 25 ppm. PMID- 2573451 TI - The effect of temperature on isolated perfused hearts of heterothermic marsupials. AB - 1. The thermal response of isolated perfused hearts of four dasyurid marsupials was determined and compared with that of two rodents. 2. Heart beat rate was strongly temperature dependent in all species. 3. The temperature of cardiac arrest in the species investigated in the present study and of others collected from the literature occurred at a mean of about 13 degrees C in homeotherms, 7 degrees C in daily heterotherms, and 1 degrees C in hibernators. 4. For both marsupials and placentals the temperature of cardiac arrest in hibernators and daily heterotherms correlated with the minimum body temperature during torpor. PMID- 2573453 TI - Metabolic rate and thermoregulation in two species of tuco-tuco, Ctenomys talarum and Ctenomys australis (Caviomorpha, Octodontidae). AB - 1. Resting metabolic rate and body temperature in function of ambient temperature were determined for two species of Ctenomys. 2. Oxygen consumption was lowest between 25 and 30 degrees C and was 0.946 +/- 0.030 and 0.968 +/- 0.022 in Ctenomys talarum (from Mar de Cobo and Necochea, respectively). Resting metabolic rate was 0.343 +/- 0.053 at 30 C in C. australis. 3. Mean rectal temperature at thermoneutrality was 36.1 +/- 0.13 and 37.3 +/- 0.17 in C. talarum and C. australis, respectively. 4. Limited thermoregulation occurred in C. talarum down to 20 degrees C but C. australis maintained body temperature down to 10 degrees C. 5. Both species of tuco-tucos became hyperthermic at ambient temperatures above thermoneutrality. PMID- 2573452 TI - Comparison between constant-protein, calorie-restricted and protein-restricted, calorie-restricted diets on growth, in vitro lipogenesis and plasma growth hormone, thyroxine, triiodothyronine and somatomedin-C (Sm-C) of young chickens. AB - 1. We studied the effects of calorie-restricted, constant-protein and calorie restricted, protein-restricted diets on growth and in vitro metabolism of male chickens from select (Cobb Line 500) and byproduct (Cobb female line) lines of broiler chickens. 2. Chickens consumed 40, 60, 80 or 100% of a prescribed formula for dietary energy (body weight in g0.70 x 16.7 kJ) in the presence of set (CEP) or varied dietary protein (VEP). 3. Chickens fed VEP were heavier (P less than 0.05) at all energy intakes than chickens fed CEP. Slope analysis of data for in vitro lipogenesis showed a significant difference between the two treatment series. 4. Plasma growth hormone was inversely related (P less than 0.05) to Sm C. Growth hormone levels were greater in chickens on a low plane of energy nutrition (40%) than on the maximum plane (100%). 5. Plasma Sm-C levels (pooled across energy series) were greater in the select than in the byproduct line. There were no differences in plasma T3 between the two lines. There was a significant increase (P less than 0.05) in T3 and a decrease in the T4/T3 ratio accompanying an increase in dietary energy. 6. Restricting dietary carbohydrate and protein compromises anabolic processes more than restricting carbohydrate alone. PMID- 2573454 TI - Liver function and protein binding in camels. AB - 1. Dehydration of camels for 10 days resulted in reduction of liver functions, expressed in longer half life and reduced clearance of bromosulfophthalein (BSP), elevated AST (ALT levels were below the limit of detection of the method) and reduced serum albumin concentrations. 2. Binding of BSP to camel serum proteins by gel permeation chromatography and by equilibrium dialysis showed very strong binding. 3. Binding parameters of various drugs to camels serum by equilibrium dialysis showed close similarities both qualitatively and quantitatively to those of humans. 4. Albumin seems to be the major serum binding protein of BSP. PMID- 2573455 TI - System A amino acid transport in a rat submandibular ductal cell line. AB - 1. Neutral amino acid transport was studied in an established cell line derived from rat submandibular glands, RSMTx. 2. The greatest portion of alpha-amino isobutyrate (AIB) transport is mediated by system A. This component is Na+ dependent, pH sensitive, markedly inhibited by methyl AIB and enhanced 2-5-fold by amino acid depletion. 3. Evidence for the presence of other neutral amino acid transport systems, presumably ASC and L, was also found in these cells. PMID- 2573456 TI - Postnatal development of the hepatobiliary transport of phenolsulfonphthalein in rats. AB - 1. The postnatal development of the biliary excretion of phenolsulfonphthalein (PSP) was studied in male Wistar rats. 2. Following i.v. injection of PSP at 200 mumol/kg body wt, a maximal biliary excretion of 175 +/- 10 nmol/min/100 g body wt and 32 +/- 5 nmol/min/100 g body wt was reached for unconjugated and conjugated PSP, respectively, in the adult group. 3. The maximal biliary excretion of conjugated PSP was significantly lower in the 20-, 30- and 40-day old groups as compared to the adults. The excretion of unconjugated dye was also significantly lower in 20- and 30-day-old rats. 4. The postnatal development of PSP excretion was unrelated to changes in the activity of UDP glucuronosyltransferase. The importance of other factors is also discussed. PMID- 2573457 TI - Resistance of mammalian red blood cells of different size to hypertonic milieu. AB - 1. The resistance of different mammalian red blood cells (RBCs) to hyperosmotic environments was studied. RBCs of six mammalian species were exposed to 10 increasingly hyperosmotic NaCl solutions for 24 hr at 5 degrees C. 2. The osmolality at which the amount of liberated haemoglobin reached a preset level (e.g. 3-4% of the total haemoglobin) showed a linear correlation with negative slope with RBC volume. This indicates that small RBCs are more resistant to hyperosmotic milieu than large ones. 3. A similar relation can be found from literature data when maximal urinary tonicities are plotted as a function of RBC volume, i.e. animals with the ability to produce highly concentrated urine have small RBCs. 4. RBC volume and maximal urinary tonicity in mammals are therefore tightly linked. Future research will have to show whether this correlation is fortuitous or not and whether, as can be speculated, RBC size is directly or indirectly regulated by the kidney. PMID- 2573458 TI - The influence of multiple photoperiods and pinealectomy on gonads, pelage and body weight in male meadow voles, Microtus pennsylvanicus. AB - 1. Chronic exposure of male Microtus pennsylvanicus to photoperiods with 8, 10 and 12 hr of light per day results in gonadal regression, molt to winter pelage and significant reductions in body weight and food consumption relative to voles kept on photoperiods with 13 and 14 hr of light per day. 2. A precise critical daylength is observed in this species as exposure to photoperiods with 12 or fewer hours of light per day results in complete gonadal involution, seasonal molt and loss of body weight. 3. Pinealectomy abolishes all short-photoperiod induced responses in this species. PMID- 2573459 TI - Comparisons of turkey embryos incubated in tenuous or dense gas environments--II. Organ growth. AB - 1. Eggs from large white turkeys were incubated in tenuous and dense gas atmospheres. 2. Tenuous gases resulted in heavier embryos until the onset of the plateau stage in oxygen consumption when dense gas environments caused heavier embryos. 3. Tenuous gases decreased heart and lung weight but increased liver weight. 4. Oxygen supplementation in tenuous gases increased liver weights but had no effect on lung or heart weights. 5. The data suggest an interaction of gas density and partial pressures of individual gases which affects breathing and the physiology of developing poult embryos. PMID- 2573460 TI - Effect of intestinal ischaemia on intestinal VIP levels and VIP interaction with intestinal epithelial cells from rat. AB - 1. The number (but not the affinity) of vasoactive intestinal peptide (VIP) receptors in small intestinal epithelial cells decreased following intestinal ischaemia in rats as compared to sham-operated animals. 2. There was a parallel decrease of the efficiency (but not the potency) of the neuropeptide upon cyclic AMP formation at the same level after intestinal ischaemia. 3. The surgical manipulation did not modify the level of VIP immunoreactivity in the gut segment studied. 4. These results suggest that the VIPergic system is not directly involved in the high loss of water and electrolytes that appears following intestinal ischaemia. PMID- 2573462 TI - The excretion of prostacyclin (PGI2) in milk and its possible role as a vasodilator in the mammary gland of goats. AB - 1. Prostacyclin production in mammary gland of two lactating goats measured as the excretion in milk of 6-ketoprostaglandin F1 alpha (6-KPGF1 alpha) was followed for 16 days before, during and after exogenous administration of recombinant bovine growth hormone (GH). 2. 6-KPGF1 alpha was detected in all milk samples in concentrations ranging from 32-99 pg/ml milk independently of the time of sampling. 3. GH-treatment significantly increased milk yield, the concentration and excretion of 6-KPGF1 alpha in milk. 4. The concentration of milk 6-KPGF1 alpha was positively correlated with milk yield in the high (R2 = 0.35), but not in the low yielding goat (R2 = 0.003). 5. The possible role of prostacyclin as a local vasodilator in the mammary gland of goats is discussed. PMID- 2573461 TI - Changes in the levels of growth hormones, insulin, cortisol, thyroxine and somatomedin-C/IGF-1, with increasing gestational age in the fetal pig, and the effect of thyroidectomy in utero. AB - 1. Blood samples were taken from 30 chronically catheterized pig fetuses in utero. Levels of growth hormone, insulin, cortisol, thyroxine and somatomedin C/IGF-1 were measured in the plasma of intact fetuses and the plasma of thyroidectomized fetuses at various gestational ages during the latter part of pregnancy. 2. Growth hormone levels were high (mean +/- SEM: 83 +/- 9 ng/ml and remained constant throughout this period. 3. Insulin levels were also constant and ranged between 4 and 14 mU/l. 4. Cortisol levels showed a general increase from 400 nmol/l at 97 days to 1200 nmol/l at term and this increase was not affected by thyroidectomy. 5. IGF-1 levels were lower than in the sows (48.0 +/- 3.0 ng/ml) and did not change throughout this period. 6. Thyroxine levels were also unchanged at about 92 +/- 4 nmol/l. 7. Thyroidectomy resulted in lower (P less than 0.001) thyroxine levels (28 +/- 3 nmol/l) but had no effect on the levels of any other hormone. PMID- 2573463 TI - Milk composition in the red-necked wallaby, Macropus rufogriseus banksianus (Marsupialia). AB - 1. Milk samples were collected throughout lactation from 10 captive red-necked wallabies. 2. The milk solids content increased throughout lactation and was accompanied by major changes in the relative proportions of protein, lipid and carbohydrate. 3. The carbohydrate fraction consisted of oligosaccharides in the first half of lactation but changed subsequently to monosaccharides. 4. The quantitative and qualitative changes observed were similar to those recorded for other macropodids. PMID- 2573464 TI - Effects of calcium availability on the release of ovine choriomammotropin from cotyledonary cells incubated in vitro. AB - 1. In this study, we examined the basal release of choriomammotropin (oCM) from monolayer cultures of cotyledonary cells obtained from ewes at different gestational ages. 2. oCM release increased with gestational age and displayed a similar profile to the concentration of oCM observed in maternal plasma. 3. Release of oCM was significantly (P less than 0.05; n = 9) increased in calcium depleted medium, and by treatment with either phospholipase C (0.125 units/ml) or KCl (50 mM). 4. The calcium antagonist MgCl (12 mM) and the calcium channel blocking agents verapamil (50 microM) and nefidipine (10 microM) all significantly stimulated oCM release. 5. These data are consistent with the suggestion that oCM release is inversely related to extracellular calcium concentration. PMID- 2573465 TI - Calcium reverses lidocaine-induced conduction block in rat fimbria in vitro. AB - 1. We have tested the effect of changed concentrations of Ca2+ upon lidocaine induced conduction block in rat fimbria. 2. With bath [Ca2+] of 0.25 mM, 0.5 mM lidocaine reduced the amplitude of the compound action potential to 20.2% +/- 2.25% of baseline (n = 5). 3. On changing the bath [Ca2+] to 4.4 mM, with no change in lidocaine concentration, the compound action potential increased by 33.5 +/- 6.5%. 4. In the absence of lidocaine, changing bath [Ca2+] had opposite effects. These results replicate findings by others in peripheral nerve. PMID- 2573466 TI - Ammonia formation in the medicinal leech, Hirudo medicinalis--in vivo and in vitro investigations. AB - 1. The excretion of N compounds was investigated in leeches fed various test solutions. 2. Ingestion was followed by a striking increase of NH3 release exhibiting a characteristic time-course. 3. The NH3 excreted resulted from the degradation of N compounds present in the test solutions. 4. Formation of NH3 from proteins was inhibited by kanamycin, but was unaffected in the case of amino acids. 5. Symbiotic microorganisms do not significantly contribute to NH3 formation. 6. Glutamate dehydrogenase and AMP deaminase are the enzymes most likely to be responsible for NH3 formation in Hirudo. PMID- 2573467 TI - Hematology and blood chemistry of chicks of white and black storks (Ciconia ciconia and Ciconia nigra). AB - 1. The hematology and blood chemistry of 15-68-day-old chicks have been studied. 2. Red cell numbers (1,900,000/mm3), hemoglobin content (11.5 g/dl) and hematocrit (37.5) were similar in both species of storks. 3. Total numbers of leucocytes (63,000/mm3) were similar in both species, whilst differential leucocyte counts were not. 4. Higher levels of plasma protein, GOT and cholesterol in HDL were found in white storks when compared with black storks but not differences were detected in plasma levels of triglycerides, total cholesterol, urea, uric acid, GPT and alkaline phosphatase. PMID- 2573468 TI - Thermoregulatory patterns of two sympatric rodents: Otomys unisulcatus and Parotomys brantsii. AB - 1. The adaptations to an arid environment in two closely related rodent species were investigated. 2. The rate of oxygen consumption (VO2), body temperature (Tb), evaporative water loss and minimal conductance in Otomys unisulcatus and Parotomys brantsii were determined under controlled conditions at ambient temperatures (Ta), ranging from 11-31 C. 3. Physiological features atypical of desert-adapted rodents include a basal metabolic rate higher than predicted by body mass, the low "lower critical temperature" and symptoms of heat stress at 31 degrees C. 4. The low Tb and wide thermoneutral zone recorded for both species are characteristic of desert rodent species. 5. These species' physiological abilities reflect their mesic phylogeny and we suggest that behaviour must play an important role in their survival in semi-arid areas. PMID- 2573469 TI - Inhibitory effect of anaesthesia with 2-phenoxyethanol as compared to MS222 on glucose release in isolated hepatocytes from rainbow trout (Salmo gairdneri). AB - 1. Glucose production by freshly isolated hepatocytes from rainbow trout was studied after anaesthesia of the animals with 2-phenoxy ethanol (2PE) or tricaine methanesulphonate (MS222). 2. At the end of the procedure, hepatic contents of glycogen, glucose, lactate, ATP, ADP, AMP, were not significantly different between the two treatments. 3. Glucose production was considerably lower for 2PE than for MS222 anaesthetized trouts. This discrepancy results probably from an inhibition of glycogenolysis, suggesting that 2PE anaesthetized animals were less stressed than MS222 anaesthetized ones. PMID- 2573470 TI - Effects of dietary carbohydrate and phenotype on thyroid hormones and brown adipose tissue locularity in adult LA/N-cp rats. AB - 1. Groups of lean and corpulent LA/N-cp rats were fed isoenergetic diets containing, 54% carbohydrate as maize starch (MS) or sucrose (SU), 20% protein, 16% mixed fats, plus other essential nutrients and fiber from 1.5-9 months of age. Final body weights of corpulent rats were 2-3 times those of their lean littermates, and were greater with SU than MS diet in both phenotypes. 2. Interscapular brown adipose tissue (IBAT) mass was greater in corpulent than lean and was greater with SU than MS diet in lean but not corpulent rats. IBAT cell diameters and adipocyte volumes were generally similar in both phenotypes, and were not markedly affected by dietary carbohydrate type. 3. Brown adipocyte locularity profiles were qualitatively similar in both phenotypes, and were morphologically indicative of thermogenic activity in both phenotypes. Locule profiles of corpulent animals contained a greater proportion of thermogenically less active types IV and V brown adipocytes than similarly fed lean animals, however, and locule distribution profiles were not influenced by diet. 4. Serum T3 concentrations were similar in both phenotypes, were greater in SU than MS lean rats and were not influenced by diet in the corpulent phenotype. In contrast, serum thyroxine concentrations and percent thyroxine uptake were not influenced by diet or phenotype. 5. These results are consistent with a partial impairment in BAT-mediated thermogenic activity in the corpulent phenotype and suggest that obesity in this strain may be due to factors other than biochemically defective brown adipose tissue thermogenesis. PMID- 2573471 TI - A field lab method to determine urine concentration in small mammals. AB - 1. The concentrations of 136 urine samples from four species of small mammals were compared using osmometry, refractometry and a colorimetric test for urea concentration. 2. To obtain a wide range of concentrations (430-3950 mOsm/kg), urine samples were collected under normal and dehydration conditions. 3. Regression analyses of paired values indicate that measurements of total solids concentration (refractometric method) permit evaluations of urine osmolality and estimations of the concentration of urea with a high degree of confidence. PMID- 2573472 TI - Effects of isoproterenol treatment on gustatory neural responses in three inbred strains of mice. AB - 1. Treatment of a beta-agonist, isoproterenol, for 5 days reduced chorda tympani responses to sucrose by about 40% of the control without affecting responses to other taste stimuli, such as NaCl, HCl and quinine HCl, in balb CrSlc mice whereas such reduction of sucrose responses was not observed in C57BL/6-CrSlc and C3H/HeSlc mice, although in the latter two strains long-lasting off-responses to quinine HCl appeared after the treatment. 2. In BALB mice, the magnitude of reduction of sucrose responses by isoproterenol increased with prolonging the treatment from 1 to 5 days, although it reached almost its maximum level by the 3 days treatment. 3. BALB mice with the removal of the submandibular glands showed slightly greater control responses of the chorda tympani nerve to sucrose than BALB mice with the sham-operation or the removal of the sublingual glands, and showed no significant reduction of sucrose responses by isoproterenol treatment. 4. These results suggest that isoproterenol probably did not act directly on sweetener receptors of taste cell membranes but affect them through the submandibular salivary system. PMID- 2573473 TI - The effects of essential fatty acid deficiency on brown adipose tissue activity in rats maintained at thermal neutrality. AB - 1. The consequences of essential fatty acid (EFA) deficiency on the resting metabolism, food efficiency and brown adipose tissue (BAT) thermogenic activity were examined in rats maintained at thermal neutrality (28 C). 2. Weanling male Long-Evans rats were fed a hypolipidic semi-purified diet (control diet: 2% sunflower oil; EFA-deficient diet: 2% hydrogenated coconut oil) for 9 weeks. 3. They were kept at 28 C for the last 5 weeks. Compared to controls, in EFA deficient rats the growth shortfall reached 21% at killing. 4. As food intake was the same in EFA-deficient and control rats, food efficiency was thus decreased by 40%. 5. Resting metabolism expressed per surface unit was 15% increased. 6. Non renal water loss was increased by 88%. 7. BAT weight was 28% decreased but total and mitochondrial proteins were not modified. 8. Heat production capacity, tested by GDP binding per BAT was 69% increased in BAT of deficient rats. 9. The stimulation of BAT was established by two other tests: GDP inhibition of mitochondrial O2 consumption and swelling of mitochondria. 10. It is suggested that the observed enhancement of resting metabolism in EFA-deficient rats is, in part, due to an activation of heat production in BAT. PMID- 2573474 TI - A study of diurnal changes in cortisol and glucose levels and FDPA activity in foals during the first 13 weeks of life and in their lactating mothers. AB - 1. In six standard-bred mares and their foals diurnal changes in the cortisol and glucose levels and in FDPA activity were studies for 13 weeks of foal life. 2. In the cortisol level diurnal rhythm was found in the 3rd, 7th and 11th week of foal life and in the 11th week of lactation in mares. 3. In mares the mean diurnal cortisol level changed from 32 ng/ml in the first week to 57 in the 11th week and in foals from 24 in the first week to 16 ng/ml in the 11th week. 4. In the glucose level no diurnal rhythm was observed. 5. In mares the mean diurnal glucose level after parturition was about 58 mg/100 ml and increased to 83-85 mg/100 ml. In foals it changed from 105 to 128 mg/100 ml. 6. In the activity of FDPA no diurnal rhythm was observed. 7. The mean activity of FDPA changed in mares from 3.3 to 4.4 U and in foals from 5.7 to 7.5 U. PMID- 2573475 TI - Anemia at the onset of winter in the meadow vole (Microtus pennsylvanicus). AB - 1. From 1981 to 1986, 6120 meadow voles (Microtus pennsylvanicus) were sampled for hematological indices in southeastern Manitoba, Canada. This survey revealed the sporadic occurrence of anemia in early winter at mean temperatures below about -5 degrees C. 2. Anemia was associated with leukocytosis and circulating normoblasts, suggesting a sudden, large blood loss. Individuals became anemic quickly, with no obvious predisposing factors. 3. Attempts were made to induce anemia by exposing voles in traps to various temperatures. Temperatures characteristic of most trapping sessions failed to induce anemia in both wild and laboratory-born voles. 4. Short-term exposure to more extreme temperatures (-20 to -30 degrees C) induced anemia. Voles lost blood through erosions of the epithelium of the glandular stomach, and developed other pathological lesions characteristic of hypothermia. 5. Although there was a strong association between cold weather and anemia, we could find no clear relationship between winter survival and winter weather. However, in 1984, extraordinarily cold temperatures were associated with anemia and a subsequent population decline. These events suggest a threshold mean daily temperature of about -15 degrees C, below which vole survival is grossly affected. 6. Deteriorating protein levels and energy reserves of small mammals in winter may make them particularly susceptible to cold stress. Hence, sporadic bouts of sustained cold may be responsible for some of the enigmatic winter declines seen in northern small mammals. PMID- 2573476 TI - Serotonin-induced chloride secretion in hen colon. Possible second messengers. AB - 1. Serotonin, 100 microM, induces a peak increase in short circuit current of about 150 microA/cm2 and in cord conductance of about 7 mS/cm2 and a more prolonged increase of 30 microA/cm2 and 1.4 mS/cm2 which lasts more than 30 min in hen colon. 2. The peak increase in short circuit current and cord conductance is due to a concomitant Cl- secretion. 3. The second messenger, which mediates Cl secretion, increases in short circuit current and cord conductance, is cyclic AMP as theophylline, 0.5 mM, increases the response in short circuit current to 1 microM serotonin from 38 +/- 5 to 78 +/- 8 microA/cm2 and in g from 1.1 +/- 0.4 to 2.0 +/- 0.3 mS/cm2. 4. Theophylline, 0.5 mM, also sensitizes the hen colon to cyclic AMP yielding an EC50 of 0.24 +/- 0.03 mM in the presence of theophylline compared with an EC50 of 2.3 +/- 0.2 mM in the absence of theophylline. 5. Manipulations of other putative second messenger systems, such as the prostaglandins/leucotrienes, the phosphoinositides and external Ca2+ or calmodulin-sensitive enzymes, did not influence the serotonin response in short circuit current and cord conductance, thus ruling out their importance as intracellular mediators. PMID- 2573478 TI - Comparative physiology of ciliary functions. Proceedings of a symposium of the 2d International Congress of Comparative Physiology and Biochemistry. Baton Rouge, Louisiana, August, 1988. PMID- 2573477 TI - Effect of Escherichia coli infection on growth and protein metabolism in broiler chicks (Gallus domesticus). AB - 1. A controlled experimental Escherichia coli infection was developed in broiler chicks. 2. Infection with E. coli significantly reduced feed intake, altered growth of the whole body, eviscerated carcass, skeletal muscles, heart and liver. Organ weight and/or the proportions of organs within the body were affected. 3. Protein accumulation in the eviscerated carcass, extensor digitorum communis and sartorius muscles was severely inhibited by infection, and to a greater extent than body weight. 4. Failure of muscle tissue to accumulate protein was associated with a significant decline in protein synthesis, when measured in vitro (-48%; P less than 0.05) and in vivo (-42%; P less than 0.001). Protein degradation also declined (-28.7%), but to a smaller extent than protein synthesis. 5. Although the infected chicks showed no viable bacteria at day 12 after infection, chicks did not reach the same body weight as controls by day 30 after infection. PMID- 2573479 TI - The role of axonemal components in ciliary motility. AB - 1. The axoneme is the detergent-insoluble cytoskeleton of the cilium. 2. All axonemes generate movement by the same fundamental mechanism: microtubule sliding utilizing ATP hydrolysis during a mechanochemical cycling of dynein arms on the axonemal doublets. 3. Structure, fundamental biochemistry and physiology of the axoneme are conserved evolutionarily, but the phenotypes of beating movements and the responses to specific cytoplasmic signals differ greatly from organism to organism. 4. A model of asynchronous dynein arm activity--the switch point hypothesis--has been proposed to account for cyclic beating in the face of unidirectional sliding. The model suggests that the diversity of beat phenotype may be explicable by changes in the timing of switching between active and inactive states of doublet arm activity. Evidence of axonemal splitting in arrested axonemes provides new support for the hypothesis. PMID- 2573480 TI - Adaptations of ciliary systems for the propulsion of water and mucus. AB - 1. The characteristics of ciliary systems are determined by the dominance of viscous effects over inertial effects. 2. The velocity of water propulsion depends on ciliary length, beat frequency, pattern of beating, the arrangement of the cilia and their co-ordination. Beating cilia influence a layer of water only two or three cilium lengths deep, with maximal velocity near the ciliary tip. 3. Mucus is propelled by the tips of short cilia that penetrate the mucus; these cilia are closely spaced on epithelia, and achieve slow propulsion that is relatively independent of load and does not require strong ciliary co-ordination. PMID- 2573482 TI - AIDS and immune dysfunction. Alternative etiologic mechanisms. PMID- 2573481 TI - Is neuroleptic dysphoria a healthy response? AB - We examined the relationship between neuroleptic-induced extrapyramidal effects, subjective neuroleptic dysphoria, and clinical outcome in acutely psychotic inpatients. Thirteen of 50 consecutively admitted patients experienced severe neuroleptic dysphoria. Dysphoric patients were less likely to comply with neuroleptic, but nine did complete neuroleptic trials. The compliant dysphoric inpatients received significantly lower neuroleptic doses compared with nondysphoric patients, and yet had equivalent overall improvement of psychotic symptoms and less severe extrapyramidal effects. Despite having fewer physical signs of akinesia, dysphoric patients were significantly more distressed from akinesia than nondysphoric. We conclude that while some neuroleptic dysphoric patients become noncompliant, others benefit from dysphoria by negotiating for lower, less toxic, and yet effective neuroleptic dosages. PMID- 2573483 TI - Dopexamine in cardiorespiratory failure. Commentary on drug research in the critically ill. PMID- 2573484 TI - Administration of dopexamine, a new adrenergic agent, in cardiorespiratory failure. AB - The short-term effects of dopexamine hydrochloride, a new synthetic adrenergic agent with predominant beta 2-adrenergic and dopaminergic properties, were studied in nine patients with inadequate cardiac output during the course of an episode of respiratory failure associated with lung infection. Dopexamine at doses up to 5 microgram/kg/min had no significant effect on arterial pressure or cardiac filling pressures, but increased cardiac index from 2.0 +/- 0.2 to 2.6 +/ 0.2 L/min.m2. Left ventricular stroke work increased from 17.3 +/- 15 to 22.1 +/ 2.4 g.m/m2 (p less than 0.01) and systemic vascular resistance index decreased from 3,792 +/- 1,035 to 2,194 +/- 823 dynes.s.cm5.m2 (p less than 0.01). The increase in cardiac output was in part related to an increase in heart rate from 91 +/- 6 to 102 +/- 7 beats/min. Under a mean inspiratory oxygen fraction of 0.48, the PaO2 decreased from 105 +/- 12 to 91 +/- 11 mm Hg (p less than 0.05) as venous admixture increased from 15.8 +/- 1.0 to 18.1 +/- 1.4 percent (p less than 0.05). Accordingly, the combination of inotropic, afterload-reducing and renal vasodilating effects of dopexamine can be useful acutely to increase cardiac output in critical conditions. However, its administration can be limited by a dose-related increase in heart rate. Dopexamine, like other catecholamines, alters blood oxygenation and increases venous admixture. PMID- 2573486 TI - Paralytic ileus responding to somatostatin therapy: first manifestation of a VIPoma. PMID- 2573485 TI - Prospective evaluation of high-dose bethanechol in investigation of esophageal chest pain. AB - We compared the value of bethanechol 80 micrograms/kg subcutaneously, acid infusion with a 0.1 normal hydrochloric acid, and edrophonium 80 micrograms/kg intravenously as provocative agents to reproduce chest pain and manometric alterations in 72 patients with noncardiac chest pain. No patient developed typical chest pain and manometric alteration with acid infusion, while five (6.9%) patients developed these changes with edrophonium and four (5.6%) with bethanechol. Only one patient developed diagnostic changes exclusively with bethanechol. All patients tested with bethanechol developed some degree of local pain or significant cholinergic symptoms, with two patients requiring atropine for relief. Side effects from edrophonium were infrequent (28% of patients tested) and did not require atropine administration. We conclude that, using the parameters of typical chest pain and the development of manometric alterations as proof of the esophageal origin of chest pain, bethanechol at 80 micrograms/kg adds little information beyond that obtainable with edrophonium. Further, the high incidence of bethanechol-related side effects at 80 micrograms/kg suggests that this dose should not be generally recommended. PMID- 2573487 TI - Active moiety of sulfasalazine. PMID- 2573488 TI - Inhibition of the metabolism of urethane in the mouse by dimethyl sulfoxide (DMSO). AB - Previous studies from this laboratory have shown that alcohols inhibit the localization of nitrosonornicotine and urethane in tissues of the mouse. Subsequent studies demonstrated that this inhibition of the localization of urethane was apparently due to an almost total inhibition of the metabolism of that compound by ethanol. We now report that dimethyl sulfoxide (DMSO) also almost completely inhibits the localization of urethane metabolites in tissues of the mouse and maintains a high concentration of urethane in blood. Since metabolism is essentially the only route of elimination of urethane in the mouse, this indicates that DMSO inhibits the metabolism of urethane. These studies lend further support to the suggestion that urethane is metabolized by either an alcohol dehydrogenase, an aldehyde dehydrogenase, or an alcohol-preferring isozyme of cytochrome P-450. These results indicate that studies on the metabolism as well as the carcinogenic activity of urethane (and possibly other chemicals) also may be affected by concurrent administration of DMSO. PMID- 2573489 TI - The pharmacokinetics of etretinate and its metabolites in the dog. AB - The pharmacokinetics and disposition of etretinate (ET) and its metabolites, etretin (ETA) and isoetretin (c-ETA), were studied in the dog. Administration of ET, ETA, and c-ETA by several routes of administration allowed the use of a physiologically based model to assess the relative contributions of absorption and presystemic metabolism to the oral bioavailability of these compounds. The large Vdss (214 +/- 228 liters/kg) of ET and terminal half-life of greater than 300 hr indicated the wide distribution and prolonged storage of ET in the tissues. After a dose of ET, its two metabolites, ETA and c-ETA, were also detectable in the plasma. The oral bioavailability of ET in the dog was 52.5% with 95.4% of the dose available for absorption from the gut lumen. Of the ET that was absorbed from the gut lumen, 26.4% and 25.5% was removed by the gut wall and liver, respectively. ETA was found to be a formation rate-limited metabolite of ET. Although the oral bioavailability of ETA could not be determined because its administration was not possible by the iv route, it appeared that only 16% of an orally administered dose entered the portal circulation from the gut. c-ETA was detected after administration of either ET or ETA. ETA and c-ETA were shown to be interconvertible metabolites, but the equilibrium of the conversion between c-ETA and ETA favored the formation of ETA. The oral bioavailability of c-ETA was 42.4%, but the liver showed little metabolic activity toward c-ETA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573490 TI - Metabolism of acrylonitrile to 2-cyanoethylene oxide in F-344 rat liver microsomes, lung microsomes, and lung cells. AB - The metabolism of acrylonitrile to the epoxide, 2-cyanoethylene oxide (ANO) was examined in rat liver microsomes, lung microsomes, and isolated enriched lung cell preparations. GC/high resolution MS was used to quantitate ANO in microsomal and cellular extracts by monitoring the fragment ion C2H3N (m/z 41.0265). The limit of detection was 0.05 pmol of ANO/0.5 microliter of standard solution, microsomal extract, or cellular extract injected onto the column, and the linear range of analysis was 0.05 to 12.5 pmol of ANO. Kinetic parameters of Vmax, V/K, and Km were calculated for microsomal ANO formation. Liver microsomes were quantitatively more active than lung microsomes on a mg of protein basis. The Vmax (pmol of ANO formed/min/mg of protein) was 666.61 for liver and 45.07 for lung microsomes. The V/K (pmol of ANO/min/mg of protein/microM) was 12.83 for liver and 0.02 for lung microsomes. The apparent Km was 51.93 microM and 1853.83 microM for liver and lung microsomes, respectively. When calculated as nmol of ANO formed/min/nmol of microsomal P-450, the Vmax for lung was equivalent to the Vmax for liver. ANO formation in the rat lung was cell specific. The rates of metabolism in the Clara cell-enriched fraction, the alveolar type II cell enriched fraction, and the cell suspension were 2.55, 0.38, and 0.67 pmol of ANO formed/min/mg of protein, respectively. No metabolism was observed in the endothelial (small) cell-enriched fraction or in the alveolar macrophages. The results suggest that the lung contributes to the metabolism and disposition of inhaled acrylonitrile. PMID- 2573491 TI - Characteristics of monooxygenase-mediated covalent binding of methoxychlor in human and rat liver microsomes. AB - The characteristics of the activation of methoxychlor by the hepatic microsomal monooxygenases and its covalent binding to microsomal proteins in human and untreated rat were compared. The Vmax of covalent binding is similar in both species, being 21 and 11 pmol/min/mg protein in human and rat, respectively. However, their Kmapp values show marked differences: 146 versus 5 microM for human and rat, respectively. These differences in Km values seem to reflect the affinities of the respective P-450s for methoxychlor in catalyzing the formation of the reactive intermediate (M*) and not the availability of acceptor binding sites. The observations that alternate substrates and inhibitors of P-450 monooxygenases inhibit covalent binding of methoxychlor to human liver microsomes, demonstrate that covalent binding is catalyzed by typical monooxygenases. Antioxidants/free radical scavengers, and sulfhydryl-containing compounds inhibit covalent binding in human liver microsomes, suggesting that the reactive intermediate is a free radical. A similar finding in phenobarbital (Pb) treated rats (Bulger, Temple and Kupfer, Toxicol. Appl. Pharmacol. 68:367, 1983) indicates that the mechanism of covalent binding in the two species is similar. Of interest is the observation with human liver samples that, despite differences in age, sex, and, probably, in diet among individuals, their rates of covalent binding of methoxychlor were similar. By contrast, there is a much higher covalent binding in the mature male rat than in the mature female or immature male or female rat, suggesting that developmentally controlled male-specific P 450s contribute to covalent binding in the adult male.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573492 TI - Pharmacokinetics and metabolism of a 4'-methylthio derivative of propranolol in the dog. AB - The purpose of this study was to examine the effect of a methylthio substituent in the metabolically most active position of propranolol, i.e. the 4'-position, on the pharmacokinetics and metabolism of this drug in the dog. The kinetics of 4'-methylthiopropranolol (MTP) were compared to those of propranolol following simultaneous iv doses of labeled drug and oral doses of unlabeled drug. MTP had a significantly larger volume of distribution and a longer half-life, and demonstrated a greater accumulation by red blood cells and cardiac conductile tissue than propranolol, effects which presumably are due to a higher lipophilicity of MTP. The greatest effect was on the oral clearance, which was substantially lower for MTP (1.6 vs. 5.5 liters/min) with an associated higher bioavailability (23.1 vs. 10.9%). Studies of MTP metabolism using radiolabeled drug showed that MTP, like propranolol, was eliminated entirely by metabolism. About 70% of the urinary radioactivity was extractable into ethyl acetate at pH 9.8 and pH 2.0. The extractable metabolites were separated by HPLC and identified by GC/MS, direct probe MS, and comparison with authentic compounds. Eleven metabolites were identified as sulfoxides and, in particular, sulfones of MTP and its N-dealkylated and subsequently deaminated glycollic and lactic acid metabolites. The nonextractable urinary radioactivity (30%) was isolated by DEAE Sephadex chromatography and identified by HPLC/MS as four glucuronic acid conjugates. In contrast to propranolol, there was no evidence of aromatic carbon oxidation for MTP. These observations suggest that the markedly decreased oral clearance of MTP compared to propranolol is due to qualitatively altered metabolism from a highly efficient aromatic carbon oxidation for propranolol to a less efficient sulfur oxidation for MTP. PMID- 2573493 TI - Age-related changes in the disposition of benzyl acetate. A model compound for glycine conjugation. AB - The in vivo metabolism and excretion of benzyl acetate (BA), a model compound for glycine conjugation, was examined in male Fischer 344 rats and C57BL/6N mice. Rats aged 3-4, 9, and 25 months received a single oral dose of either 5 or 500 mg/kg 14C-BA, while male mice aged 2, 13, and 25 months received a single oral dose of 10 mg/kg 14C-BA. Urine and feces were collected for 96 hr. Biliary excretion and plasma elimination were also examined in male Fischer rats after iv administration of 5 mg/kg 14C-BA. In both young and old rats and mice, hippuric acid (HA) was the major urinary metabolite after oral dosing of BA. No significant age-related difference was observed in rats in the urinary elimination of BA-derived radioactivity or in the percentage of the total dose excreted as hippuric acid (approximately 95%). Twenty-five-month old rats excreted a significantly higher percentage of the total dose as benzyl mercapturic acid (approximately 2%) than did 3- to 4-month-old rats (approximately 1%) at the 5 mg dose. Benzyl mercapturic acid excretion in 3- to 4 month-old rats was also increased significantly at 500 mg/kg BA vs. 5 mg/kg BA. Fecal excretion of BA-derived radioactivity declined significantly in 25-month old rats at both the 5 and 500 mg dose. This decrease was reflected by an age related decline in biliary excretion and higher plasma levels of BA-derived radioactivity. Examination of plasma metabolites revealed a significantly higher level of HA and benzoyl glucuronide in 25-month rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573495 TI - Disposition, metabolism, and excretion of U-71038, a novel renin inhibitor peptide, in the rat. AB - The in vivo fate of U-71038 (Boc-Pro-Phe-N-MeHis-Leu psi [CHOHCH2] Val-Ile (aminomethyl)pyridine), a potent renin inhibitor, was investigated in rats by single iv administration of tritium-labeled drug at a dose level of 5 mg/kg. The plasma concentrations of drug-related radioactivity diminished very rapidly during the first hour after dosing, with the initial concentrations measured at 2 min falling by more than 95% during the first 30 min. Estimates of the approximate half-life of this earliest phase of the plasma concentration-time curve gave an average value of 4 min. The residual amount of radioactivity after 30 min was cleared from the plasma more slowly, with trace levels still detected 48 hr after dosing. The radioactivity was recovered chiefly (91% of the dose) in feces, indicating biliary clearance as the primary route of elimination from systemic circulation. Urinary recoveries averaged 4% of the dose. Radio-HPLC profiling of plasma, urine, and bile extracts detected only a single radioactive drug-related component in these samples. Preparative HPLC was used to isolate this component from bile; mass spectral comparison to U-71038 confirmed its identity as the unchanged drug. Therefore, U-71038 does not undergo significant systemic metabolism in this species and is eliminated in bile and urine in intact form. Distribution of drug-related radioactivity was very rapid to most of the organs and tissues that were sampled, with the exception of very limited penetration into the central nervous system. Highest tissue levels of tritium were generally found in organs associated with elimination (liver, intestine, kidney) and in thyroid. PMID- 2573494 TI - Metabolic conversion of 2-propylpentanal acetals to valproic acid in vitro. Model prodrugs of carboxylic acid agents. AB - As part of an investigation to test the feasibility of using acetals as precursors of acidic drugs, the dimethyl, diethyl, [2H10]diethyl, and diisopropyl acetals of 2-propylpentanal were synthesized and their metabolic conversion to the anticonvulsant, valproic acid (2-propylpentanoic acid), was investigated. The acetals were incubated with either 10,000g supernatant or microsomes isolated from rat liver. Data from the measurement of the metabolite, valproic acid, with selected ion monitoring gas chromatography/mass spectrometry indicated that the dimethyl, diethyl, [2H10]diethyl, and diisopropyl acetals were substrates. The amount of valproic acid produced from the incubation of 2-propylpentanal diethyl acetal with 10,000g supernatant was reduced by the cytochrome P-450 inhibitor, SKF-525A. The production of acid was also decreased by lack of NADPH or oxygen. These data are consistent with a cytochrome P-450 mediated reaction. 2-Propyl-1 pentanol was the major metabolite identified from microsomal preparations free of soluble fraction enzymes. A deuterium isotope effect calculated as the ratio of the amount of valproic acid produced from unlabeled and [2H5]ethyl-labeled substrate was 1.2. Failure to detect an ester as a metabolite of 3-phenylpropanal diethyl acetal along with the results of the isotope effect studies suggest that the mechanistic pathway of acyclic acetal metabolism involves oxidation of an ether methylene and not oxidation at the acetal carbon. PMID- 2573496 TI - Human and dog, but not rat, isolated hepatocytes have decreased foreign compound metabolizing activity compared to liver slices. AB - A comparison has been made of the metabolism of biphenyl by isolated hepatocytes and liver slices from rat, dog, and human. Hepatocytes were prepared by low Ca2+ and enzyme digestion of the perfused liver of rat or liver slices from the rat, dog, and human. The ratio of free to total hydroxybiphenyl formation (R) was a sensitive measure of hepatocyte functional viability in perfusion-isolated rat hepatocytes, showing a significant negative correlation (r = -0.920, p less than 0.01) with trypan blue exclusion (TBE). Rs for rat hepatocytes prepared by the perfusion and slice-digestion techniques were not significantly different. Biphenyl metabolism and TBE in rat, dog, and human hepatocytes isolated by the slice-digestion technique were compared. Total hydroxybiphenyl formation by dog and human hepatocytes was 21% and 4% of that seen with rat hepatocytes. Rs for rat, dog, and human hepatocytes were 0.19, 0.46, and 0.63, respectively. TBE for all the hepatocyte preparations was approximately 90%. In contrast to the hepatocytes, total hydroxybiphenyl formation by slices of dog and human liver was 106% and 108%, respectively, of that seen with slices of rat liver. Rs for rat, dog, and human liver slices were 0.11, 0.21, and 0.26, respectively. These results suggest that hepatocytes prepared by the slice-digestion technique from dog and human but not rat liver have lost some of their ability to oxidize biphenyl and form biphenyl conjugates. This may be due to damage to the hepatocytes during isolation. TBE does not appear to be an accurate measure of hepatocyte functional viability between species. It is concluded that liver slices may provide a better model than isolated hepatocytes for foreign compound metabolism studies with dog and human liver. PMID- 2573497 TI - Biotransformation of tipredane, a novel topical steroid, in mouse, rat, and human liver homogenates. AB - The in vitro biotransformation pathways of 3H-tipredane (3H-TP) were studied. 3H TP, at concentrations of 1 and 250 microM, was incubated with the 10,000g supernatant fraction of the liver homogenates of mice, rats, and one human. The incubation mixtures were deproteinated with methanol and, after removal of methanol by evaporation, extracted with dichloromethane. The dichloromethane extracts were then fractionated by HPLC. 3H-TP was extensively biotransformed by the liver homogenates of the three species studied; 17 metabolites were isolated and characterized by their retention times on HPLC compared to those of the reference standards. Fourteen metabolites were identified using MS and, for some, NMR spectroscopy. Three major biotransformation pathways of TP were identified: 1) sulfoxidation, 2) elimination of the alkylthio groups, and 3) hydroxylation of the steroid nucleus. Combinations of these processes and subsequent reactions resulted in the formation of numerous metabolites whose biological activities were significantly less than that of TP. The separation of local anti inflammatory activity from systemic side effects observed for TP in animals and humans is most probably due to its metabolic inactivation, primarily in the liver. PMID- 2573498 TI - Metabolism and disposition of the 5-hydroxytryptamine uptake blocker sertraline in the rat and dog. AB - Sertaline [1S,4S-N-methyl-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydro-1- naphthalenamine] is a potent and selective inhibitor of neuronal serotonin uptake and is currently under development for the treatment of depression and of obesity. The drug is greater than 97% bound to plasma proteins, yet extensively distributes into tissues. The whole brain concentration of sertraline in the rat is more than 40-fold higher than that in plasma, and the volume of distribution is about 25 liters/kg in the rat and dog. Sertraline is extensively metabolized by the rat and dog prior to excretion. The metabolic clearance of sertraline is greater than 35 ml of blood/min/kg in each species, and first-pass metabolism occurs with oral administration. Initial metabolic steps include N-demethylation, N-hydroxylation, oxidative deamination, and glucuronidation of sertraline carbamic acid, which in solution is in equilibrium with sertraline and carbon dioxide. The N-desmethyl metabolite, which is 10-fold less potent as an inhibitor of serotonin uptake, is formed in both species. Plasma AUC for desmethyl sertraline is 66 to 270% of that for sertraline, and is dependent on the species examined and route of drug administration. Sertraline and desmethyl-sertraline undergo oxidative deamination to the corresponding ketone, which is subsequently hydroxylated at the alpha-carbon, forming a diastereomeric metabolite pair. The glucuronides of sertraline carbamic acid, N-hydroxy sertraline, and the alpha hydroxy ketone diastereomers comprise 45% and 82% of the total radiolabel excreted in urine and bile of bile duct-cannulated rats and dogs, respectively. Bile is the major route of elimination in both species. PMID- 2573499 TI - Cytochromes P-450 in murine lung. An immunohistochemical study with monoclonal antibodies. AB - We have used a panel of monoclonal antibodies (Mabs) and immunofluorescence to investigate the distribution of species of cytochrome P-450 inducible by 3 methylcholanthrene (3-MC) and phenobarbital (PB) in the lungs of "responsive" C57BL/6 and "nonresponsive" DBA/2 mice. A Mab (1-7-1) specific for an epitope common to rat cytochromes P-450c (P450IA1) and P-450d (P450IA2) detected the corresponding murine species (P1-450/P3-450) in cells of the alveolar septa, including type II cells, as well as endothelial cells lining blood vessels. Cytochrome P1-450 (Mabs 1-31-2, 1-36-1) is localized in type II cells of the alveolar septa but is not found in endothelial cells. Patterns of immunoreactivity with the 3-MC-specific Mabs indicate that alveolar septal cells possess P1-450 and P3-450, whereas endothelial cells of the vasculature are not the sites of P1-450, but rather of P3-450. Similar immunoreactivities for the 3 MC-defined P-450s are demonstrated in the lungs of C57BL/6 and DBA/2 mice. A Mab (2-66-3) which recognizes P-450b/P-450e (P450IIB1/P450IIB2) is reactive with P 450s localized in type II alveolar and bronchiolar epithelial cells, including nonciliated Clara cells. The epitope for cytochromes P-450b/P-450e is present in constitutive form within the lung and does not appear to be responsive to induction by PB. In contrast, epitopes for P1-450/P3-450 and P1-450 are not expressed constitutively but are only detected when induced by 3-MC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573500 TI - First-pass metabolism of salicylamide. Studies in the once-through vascularly perfused rat intestine-liver preparation. AB - Salicylamide (SAM) metabolism was studied in a once-through in situ perfused rat intestine-liver preparation in a manner which mimicked the first-pass effect. SAM (40 or 200 microM) was delivered into the intestine via the superior mesenteric artery at a flow rate of 7.5 ml/min. The intestine venous outflow into the portal vein and the hepatic arterial flow (2.5 ml/min; without drug) served as dual inflows into the liver. The steady state intestinal and hepatic extraction ratios were 0.262 +/- 0.055 and 0.992 +/- 0.014, respectively, at 40 microM, and 0.206 +/- 0.035 and 0.638 +/- 0.117, respectively, at 200 microM. SAM glucuronide was found to be the only metabolite formed by the intestine at both doses. Less than 3% of the dose was secreted into the intestinal lumen, with SAM glucuronide and SAM as the major and minor components, respectively. Hepatic metabolism of SAM, however, revealed SAM sulfation as the predominant pathway, while glucuronidation and hydroxylation were minor metabolic pathways. About 6% of the dose was excreted into bile, mostly as SAM and gentisamide glucuronides. The interrelationship between the intestine and liver clearances was also examined by mass balance considerations and simulation of data. The total rate of elimination of substrate across the two organs is the sum of the rates of metabolism by each organ. However, the overall effective extraction ratio and, hence, the clearance, are less than the sum of the individual extraction ratios and organ clearances, respectively. Our results showed that intestinal metabolism regulated the available substrate for hepatic elimination, and hence modified the contribution of hepatic metabolism in the overall first-pass effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573501 TI - Deimidazolation of antimycotic croconazole through epoxide formation in rabbit liver microsomes. AB - Incubation of croconazole with rabbit microsomes in the presence of NADPH + 18O2 produced the major metabolite, 2-(3-chlorobenzyloxy)phenacyl alcohol (M2) containing one 18O atom. Also detected in the ethyl acetate extracts from the incubation mixture was the remainder of the product, imidazole. These results indicate that the deimidazolation process from croconazole involves the intermediate, croconazole epoxide, which is hydrolyzed immediately to M2 and imidazole. This reaction is mediated by cytochrome P-450 as indicated by the requirement of NADPH, the incorporation of 18O, and inhibition by 10 mM metyrapone, 0.1 mM SKF 525-A, and CO/O2 (50/50, 80/20). Double reciprocal plots of M2 formation give a straight line, suggesting that the reaction may be mediated by an isozyme of cytochrome P-450 families. PMID- 2573502 TI - In vitro studies of human liver alcohol dehydrogenase variants using a variety of substrates. AB - Alcohol dehydrogenase (ADH) is genetically polymorphic, and large differences in allele frequencies exist between the major human races. Genetic variants at the ADH2 gene locus include the beta 2-ADH ("atypical" ADH) present in 85% of Orientals and the beta 1-ADH ("normal" ADH) present in 85 to 95% of whites. Although the presence of one or the other of these ADH variants does not significantly affect the rate of ethanol oxidation in the living subject, it may affect that of other substrates. The overall objective of this work was to screen in vitro for ADH substrates which might be differentially metabolized by these ADH2variants in living subjects. In an in vitro screening method using autopsy livers at pH 8.5, the formation or disappearance of NADH at 340 nm was measured before and after exposure to 4-methylpyrazole, an ADH-specific competitive inhibitor. The screening test revealed three new substrates and suggested that alcohol substrates fall into two groups. The majority of substrates belonged to a group which was oxidized at a significantly lower rate by the beta 2-ADH as compared to the beta 1-ADH, but this was not the case for a small group which included ethanol. Subsequent kinetic studies of selected alcohols tended to indicate a uniqueness of ethanol kinetics in that both KM and Vmax favored its oxidation by beta 2-ADH rather than by the beta 1 variant. None of the other seven tested alcohols showed a similar differential. Also, reduction of aldehydes and ketones tended to be moderately slower by beta 2- than by beta 1 ADH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573503 TI - Systemic intestinal metabolism of 1-naphthol. A study in the isolated vascularly perfused rat small intestine. AB - Using a vascularly and luminally perfused rat small intestine, we studied the systemic intestinal metabolism of the model substrate 1-naphthol (1-N) to 1 naphthol-beta-D-glucuronide (1-NG). An intestinal extraction ratio of 0.30 +/- 0.02 was found for 1-N. This implies that intestinal metabolism represents up to 14% of the total plasma clearance of 1-N in the rat in vivo. The formed 1-NG was preferentially released into the vascular perfusate, suggesting specialized transport carriers for 1-NG in brushborder and basolateral membrane. When the vascular flow rate was decreased from 10 to 0.5 ml/min, the clearance of 1-N appeared to be completely flow dependent. The apparent conflict between a low extraction ratio (0.30 +/- 0.02 at all flows investigated) and a flow rate limited 1-N clearance can be explained by the presence of an intestinal vascular bed with a high extraction ratio. We suggest that 1-N is completely extracted from the mucosal blood flow. This view was confirmed by the results of experiments in which the capillary flow of the intestinal mucosa was decreased by infusion of noradrenaline. As a result a temporary decrease in the 1-N extraction ratio was observed. The contribution of the intestine to the total clearance can be masked by the hepatic clearance, because the blood supply to the intestine and liver is coupled in series. An equation was derived to describe the relative contribution of the intestine to the mesenteric clearance of the intestine-liver system. It appears that the effective contribution of the intestine to the mesenteric clearance is of little interest for high extraction drugs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573504 TI - The influence of phenobarbital administration on hepatic monooxygenase activity at various stages of gestation in the rat. PMID- 2573507 TI - [Mechanisms of molecular resistance to cancer chemotherapy]. PMID- 2573506 TI - Rational use of antihistamines in allergic dermatological conditions. AB - For many years, the use of antihistamines in dermatological conditions has been closely linked to the treatment of urticarias and to the symptomatic relief of pruritus. H1-receptor antagonists are the first-line drugs for urticarias. Those urticarias of type I immunological origin respond better than physical urticarias. H2-receptor antagonists may be added for refractory patients but are rarely effective alone. Conventional H1-blockers are frequently associated with somnolence and anticholinergic effects. Therefore, new antihistamines without depressive effects on the central nervous system have recently been introduced. In other pruritic conditions such as atopic dermatitis the limited effects of these drugs suggest that histamine is not the only mediator involved in pruritus. In these cases, their beneficial effects seem to be due mainly to their sedative properties; recently available nonsedative H1-blockers are less effective. PMID- 2573508 TI - [Management of insomnia]. PMID- 2573509 TI - Somatostatin inhibition of growth hormone secretion by somatotropes from male, female, and androgen receptor-deficient rats: evidence for differing sensitivities. AB - To investigate the role of somatostatin (SRIF) in regulating sexually dimorphic GH secretion, we used a reverse hemolytic plaque assay and acutely dispersed somatotropes from age-matched normal male, normal female, and androgen receptor deficient, testicular feminized (Tfm) rats. Hemolytic plaques were developed after a 90-min incubation in the presence of GH antiserum, 10 nM GH-releasing hormone (GHRH), and the following concentrations of SRIF: 0, 0.001, 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3, 10, 30, and 100 nM. Additional studies were performed with 0 or 100 nM SRIF in the absence of GHRH. The absolute number of somatotropes (x10(6); mean +/- SEM) recovered from the pituitaries of Tfm rats (1.73 +/- 0.18) was significantly greater than that from the males (1.11 +/- 0.13; P = 0.01); the number from female rats (1.30 +/- 0.15) was not different from that of either male or Tfm animals. GHRH-stimulated GH secretion, as estimated by the mean GH plaque area (micron2 x 10(4); mean +/- SEM) in the absence of SRIF, was greater for somatotropes from male rats (3.36 +/- 0.41) than that for either Tfm (2.27 +/ 0.32; P = 0.02) or female (1.78 +/- 0.24; P = 0.001) rats; values for the latter two groups did not differ. However, mean GH plaque areas for each group during maximal SRIF inhibition in either the presence or absence of GHRH were indistinguishable from each other and from mean plaque areas obtained under basal conditions. As demonstrated by a lesser EC50 value (0.04 +/- 0.02 nM; mean +/- SEM), somatotropes from female rats were more sensitive to the inhibitory effect of SRIF than were those from either male (EC50 = 1.82 +/- 0.45; P = 0.0001) or Tfm (EC50 = 0.74 +/- 0.22, P = 0.0001) rats; values for the latter two groups were indistinguishable. These observed differences suggest that gender and/or the gonadal hormone environment may be important determinants of the inhibitory effects of SRIF on GH secretion by the somatotrope. While these gender-associated differences may represent effects of the gonadal hormones directly on the somatotrope, they could reflect modulation of the secretion of hypothalamic SRIF and/or GHRH by the prevailing gonadal hormone environment. Such gender-related differences may contribute to the overall sex-dependent patterns of GH secretion in the intact animal. PMID- 2573510 TI - In the rat, interleukin-1 alpha and -beta stimulate adrenocorticotropin and catecholamine release. AB - Recombinant interleukin-1 alpha and -beta (Il-1 alpha and -beta) have been evaluated for their abilities to stimulate ACTH and catecholamine secretion in the intact adult male rat. Additionally, the role of adrenergic-dependent pathways in mediating Il-1-induced ACTH release has been assessed. The iv or intracerebroventricular injection of either Il-1 alpha or Il-1 beta caused dose related increases in plasma ACTH, epinephrine, and norepinephrine levels. While at low iv doses (less than or equal to 10 ng), Il-1 beta was more effective than Il-1 alpha at releasing ACTH, no measurable differences were noted at higher doses. In contrast, Il-1 beta was significantly more active at all doses in elevating plasma ACTH levels after intracerebroventricular injection. Similarly, Il-1 alpha was more effective than Il-1 beta at stimulating epinephrine, but not norepinephrine, secretion after icv injection. Because of the ability of catecholamines to alter ACTH secretion, we then examined the role of adrenergic dependent pathways as possible mediators of Il-1-induced ACTH secretion. Blockade of adrenergic receptors by the concomitant administration of prazosin and propanolol prevented the simultaneous actions of phenylephrine and isoproterenol on the corticotrophs, but did not significantly alter the effect of peripherally administered Il-1 alpha on ACTH release. These results suggest that both Il-1 alpha and Il-1 beta stimulate ACTH and catecholamine secretion in the rat. Despite the ability of the lymphokine to elevate plasma epinephrine and norepinephrine values, circulating catecholamines do not appear to represent essential modulators of ACTH release induced by acutely injected Il-1. PMID- 2573505 TI - Famotidine. An updated review of its pharmacodynamic and pharmacokinetic properties, and therapeutic use in peptic ulcer disease and other allied diseases. AB - Famotidine is a highly selective histamine H2-receptor antagonist. In healthy volunteers and patients with acid hypersecretory disease it is approximately 20 to 50 times more potent at inhibiting gastric acid secretion than cimetidine and 8 times more potent than ranitidine on a weight basis. As shown in placebo controlled trials, famotidine is effective in healing both duodenal and gastric ulcers. Famotidine 20mg twice daily or 40mg at bedtime achieves healing rates and symptom relief similar or superior to those achieved by cimetidine 800mg daily or ranitidine 300mg daily in patients with peptic ulcer disease. Results of 1 placebo-controlled study suggest that famotidine prevents recurrence of duodenal ulcer, but comparative trials are needed to establish its relative efficacy in maintenance therapy. The few non-comparative trials conducted to date also suggest that famotidine 10 to 20mg twice daily may be effective in the treatment of gastritis and reflux gastro-oesophagitis. In comparative trials, famotidine was similar in efficacy to cimetidine in the treatment of upper gastrointestinal bleeding and to ranitidine in the prevention of pulmonary aspiration of acid. In patients with Zollinger-Ellison syndrome, the potency and long duration of action of famotidine may confer an advantage over other H2-receptor antagonists--in individualised doses (mean 0.33 g/day) famotidine successfully controlled acid secretion for up to 72 months in 1 study of such patients. Accumulated clinical evidence confirms that famotidine is very well tolerated and is free of the antiandrogenic effects infrequently reported with cimetidine. Moreover, famotidine is not associated with altered hepatic metabolism of drugs. Thus, famotidine is an effective, well-tolerated alternative to cimetidine and ranitidine. Famotidine is also promising as maintenance therapy for preventing recurrence of duodenal ulcer and as initial or maintenance treatment of gastric hypersecretory disorders, but further clinical experience, particularly in the long term, is needed to define the relative efficacy and tolerability of famotidine in these indications. PMID- 2573511 TI - Molecular cloning of murine intercellular adhesion molecule (ICAM-1). AB - We have previously reported a murine lymphocyte surface antigen MALA-2 of approximately 95,000 Mr which is expressed mainly on activated lymphocytes. The rat monoclonal antibody YN1/1 that detects this antigen profoundly inhibits mixed lymphocyte response. We have now purified MALA-2 and determined its partial amino acid sequence. By using non-redundant synthetic oligonucleotides as probes, based on the amino acid sequence, we have isolated two full length cDNA clones encoding MALA-2. The two clones are identical except for the 5' end sequence. Expression of MALA-2 on transfected COS cells is only achieved with one of the two cDNA clones. The nucleotide sequence as well as the deduced amino acid sequence of MALA-2 display striking homology with those of the recently reported human intercellular adhesion molecule ICAM-1. All the unique features of the human ICAM 1, including its homology with the neural adhesion molecule NCAM, its internal repeat structure and the immunoglobulin-like structure, are found in MALA-2. Furthermore, purified MALA-2 crosslinked to a solid support binds Con A blasts that express LFA-1, the putative receptor for ICAM-1, and the binding can be blocked by YN1/1 antibody or antimurine LFA-1 antibody indicating a direct interaction of these molecules in cell adhesion. Therefore, we consider MALA-2 to be the murine homolog of human ICAM-1. Since ICAM-1 is known to be of primary importance in immune responses and inflammatory reactions, having a monoclonal antibody and a mouse model will provide the opportunity to study the functional role of ICAM-1 in vivo. PMID- 2573512 TI - Site-specific mutagenesis identifies amino acid residues critical in prohormone processing. AB - Peptide hormones are generally synthesized as inactive higher mol. wt precursors. Processing of the prohormone into biologically active peptides by specific proteolytic cleavages occurs most often at pairs of basic amino acids but also at single arginine residues. To study the role of protein secondary structure in this process, we used site-directed mutagenesis to modify the predicted secondary structure around the cleavage sites of human prosomatostatin and monitored the processing of the precursor after introduction of the mutated cDNAs in Neuro2A cells. Amino acid substitutions were introduced that affected the possibility of forming beta-turn structures in the immediate vicinity of the somatostatin-28 (S 28) and somatostatin-14 (S-14) cleavage sites. Infection of Neuro2A cells with a retrovirus carrying a human somatostatin cDNA resulted in the expression of prosomatostatin and its processing into S-28 and S-14, indicating that these cells have the necessary enzymes to process prohormone at both single and paired amino acid residues. Disruption of the different beta-turns had various effects on prosomatostatin processing: substitution of Ala for Pro-5 drastically decreased prosomatostatin processing and replacement of Pro-9 by Ala led to the accumulation of the intermediate maturation product [Arg-2Lys-1]-S-14. In contrast, substitution of Ala for Asn-12, Gly+2 and Cys+3 respectively had only very little effect on the proteolytic processing of prosomatostatin. Our results show that amino acids other than the basic amino acid residues are required to define the cleavage sites for prohormone proteolytic processing and suggest that higher orders of protein structure are involved in substrate recognition by the endoproteases. PMID- 2573513 TI - Human herpes virus-6 increases HIV-1 expression in co-infected T cells via nuclear factors binding to the HIV-1 enhancer. AB - Human Herpes virus-6 (HHV-6) can co-infect with HIV-1 human CD4+ T-cells, leading to accelerated cell death, and factors in HHV-6-infected cells stimulate HIV-1 LTR directed gene expression. In this study, we have examined the mechanism of HIV-1 activation by HHV-6 and localized the cis-acting sequences of HIV-1 LTR responsive to trans-activation. Increased HIV-1 LTR directed gene expression is obtained in HIV-1 infected cells co-infected with HHV-6, or in HHV-6 infected cells co-transfected with the HIV-1 tat gene. Parallel increases of HIV-1 specific transcripts are seen by in situ hybridization in HHV-6/HIV-1 doubly infected cells as compared to single HIV-1 infection. Similarly, infection by HHV 6 increases the steady-state level of HIV-1 LTR mRNA that parallels CAT enzymatic activity, suggesting a transcriptional and/or post-transcriptional activation. Sequences necessary for HIV-1 LTR activation by HHV-6 are distinct from those required for that tat response and map to a region of the HIV-1 LTR from -103 to 48. The HIV-1 enhancer sequence (-105 to -80) is sufficient to confer HHV-6 inducibility to a heterologous promoter, and nuclear protein(s) activated or induced by HHV-6 infection specifically bind to the NF kappa B motifs of the HIV 1 enhancer region.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573514 TI - A post-ribosomal supernatant from activated Xenopus eggs that displays post translationally regulated oscillation of its cdc2+ mitotic kinase activity. AB - A cell-free extract prepared from activated Xenopus eggs by high-speed centrifugation displays one spontaneous cycle of activation and inactivation of histone H1 kinase and MPF activity that is largely attributable to Xenopus p32cdc2. The timing of the oscillation closely follows that observed in intact eggs, is associated with large changes in endogenous protein phosphorylation and depends entirely on post-translational events. The extract can be fractionated into soluble and particulate material, both of which components are required for the oscillatory behaviour. Kinase activation does not require Mg+ ATP, but its rapid inactivation, which coincides with the destruction of cyclin, is inhibited both by EDTA and the protein kinase inhibitor 6-dimethylaminopurine. This suggests that protein phosphorylation is required for cyclin destruction and kinase inactivation. PMID- 2573515 TI - Tissue-specific and hormonal regulation of angiotensinogen minigenes in transgenic mice. AB - Angiotensinogen is the precursor of the potent vasoactive peptide angiotensin II, and is therefore an important determinant of blood pressure and electrolyte homeostasis. In order to map the tissue-specific and inducible enhancer elements governing angiotensinogen gene expression in transgenic mice, we constructed minigenes containing either 0.75 kb or 4 kb or 5' flanking DNA from the BALB/c angiotensinogen gene. Sequences necessary and sufficient to mediate induction by glucocorticoids, oestrogen and bacterial endotoxin were contained on the minigene bearing 0.75 kb of DNA upstream of the capsite. This construct was also able to confer tissue specificity in the majority of organs producing angiotensinogen. In the testis and salivary gland, differences between the donor (BALB/c) and recipient (Swiss) strains were responsible for the apparently aberrant expression of the minigene constructs. The genetic lesion responsible for these expression polymorphisms has been characterized using recombinant inbred mice. An EcoRI restriction fragment length polymorphism which co-segregates with the angiotensinogen expression phenotypes into many inbred mouse strains is also described. PMID- 2573516 TI - Isolation of two tissue-specific Drosophila paired box genes, Pox meso and Pox neuro. AB - Two new paired domain genes of Drosophila, Pox meso and Pox neuro, are described. In contrast to the previously isolated paired domain genes, paired and gooseberry, which contain both a paired and a homeo-domain (PHox genes), Pox meso and Pox neuro possess no homeodomain. Evidence suggesting that the new genes encode tissue-specific transcriptional factors and belong to the same regulatory cascade as the other paired domain genes includes (i) tissue-specific expression of Pox meso in the somatic mesoderm and of Pox neuro in the central and peripheral nervous system, (ii) nuclear localization of their proteins, (iii) dependence on prd activity and (iv) presence of the paired domain in genes of known regulatory activity. While no mutant phenotypes of Pox meso and Pox neuro have yet been discovered, a murine gene with a paired domain closely homologous to that of Pox meso has recently been identified with the undulated mutant. Both Pox meso and undulated are expressed in tissues derived from the somatic mesoderm. The five known Drosophila paired domains fall into three classes: (i) the prd,gsb-class, (ii) the Pox meso, undulated-class and (iii) the Pox neuro class which probably includes the paired domain of the murine gene Pax 2. PMID- 2573517 TI - Effects of mutations in heat-shock genes groES and groEL on protein export in Escherichia coli. AB - Escherichia coli heat-shock proteins GroES and GroEL are essential cytoplasmic proteins, which have been termed 'chaperonins' because of their ability to assist protein assembly of bacteriophage capsids and multimeric enzymes of foreign origin. In this report we show that temperature-sensitive mutations in groES and groEL genes cause defective export of the plasmid-encoded beta-lactamase (Bla) in vivo. Since efficient translocation of proteins across biological membranes is thought to be supported by cytoplasmic factors that protect presecretory molecules from being misfolded, these results suggest that both GroES and GroEL proteins possess a chaperone function by which they facilitate export of Bla. The translocation of other secretory proteins, however, appears to depend minimally on GroE, suggesting that GroE interacts only with a specific class of secreted proteins. PMID- 2573518 TI - Cloning and expression in Escherichia coli of Haemophilus influenzae fimbrial genes establishes adherence to oropharyngeal epithelial cells. AB - In this report the first example of functional expression of a fimbrial gene cluster of a non-enteric human pathogen in Escherichia coli is described. This is shown for Haemophilus influenzae fimbriae which mediate adherence to oropharyngeal epithelial cells. A genomic library of H.influenzae type b, strain 770235f+bo, was constructed using a cosmid vector and screened with a synthetic oligonucleotide probe derived from the N-terminal sequence of the fimbrial subunit of H.influenzae. Four cosmid clones were found which hybridized to this oligonucleotide probe. Escherichia coli strains harbouring these clones expressed the H.influenzae fimbriae at their cell surface, as was demonstrated in a whole cell ELISA and by immunogold electron microscopy using a monoclonal antibody specific for the H.influenzae fimbriae. Surface expression could be maintained during subcloning until a minimal H.influenzae DNA insert of approximately 8.1 kb was obtained. Escherichia coli strains harbouring the 8.1 kb H. influenzae DNA were able to cause a mannose-resistant adherence to oropharyngeal epithelial cells and a mannose-resistant haemagglutination of human AnWj-positive erythrocytes. The nucleotide sequence of hifA, the gene encoding the major fimbrial subunit, was determined. The predicted amino acid sequence shows a significant homology with a number of E.coli fimbrial subunits. PMID- 2573519 TI - Elimination of CD8+ thymocytes in transgenic mice expressing an anti-Lyt2.2 immunoglobulin heavy chain gene. AB - Individual T cell populations are characterized by specific surface proteins, namely by the T cell receptor complex (TCR) and by two accessory molecules, CD8 (Lyt2) and CD4 (L3T4). CD8 and CD4 are required for T cell interactions with class I or class II major histocompatibility complex molecules. In the thymus, immature CD8(-4)-TCR- cells differentiate, possibly via a short stage of CD8+4- thymocytes, into CD8+4+ TCR+ T cells and mature further into the main T cell populations, the CD8+4- TCR+ cytotoxic T lymphocytes and the CD4+8- TCR+ T helper cells. In order to analyse the differentiation steps involving CD8, we generated transgenic mice expressing mu heavy chain genes from an anti-Lyt2.2 hybridoma. Transgenic lines expressing either the complete (mu sm) or only the secreted mu protein (mu s) suffer from a severe depletion of their CD8+4+ thymocytes affecting also the mature CD8+4- and CD4+8- populations. The depletion is correlated to the expression of transgenic mu-chain proteins within thymocytes. This intrathymocyte expression of the mu chain prevents CD8-4- thymocytes from further differentiation, most probably via intracellular interactions between mu heavy chain and CD8 proteins. These results show that CD8 plays an important role during thymocyte maturation. PMID- 2573520 TI - Genetic and molecular mapping of the Hmt region of mouse. AB - We have mapped a new region of the mouse major histocompatibility complex (MHC) that contains the nuclear gene, Hmt, for the maternally transmitted antigen, Mta. The Hmt region of chromosome 17 lies between a recombinational breakpoint distal to Tla and another proximal to Tpx-1, thus including Pgk-2. A novel MHC class I gene fragment, R4B2, was cloned and mapped to this region as was another new class I gene, Thy19.4. Both lie proximal to Pgk-2, within the distal inversion in t-haplotypes. The presence of several other MHC class I genes in the Hmt region is predicted from analysis of the recombinants that define the region. PMID- 2573521 TI - Multiple replication factors augment DNA synthesis by the two eukaryotic DNA polymerases, alpha and delta. AB - DNA synthesis by two eukaryotic DNA polymerases, alpha and delta, was studied using a single-strand M13 DNA template primed at a unique site. In the presence of low amounts of either DNA polymerase alpha or delta, DNA synthesis was limited and short DNA strands of approximately 100 bases were produced. Addition of replication factors RF-A, PCNA and RF-C, which were previously shown to be required for SV40 DNA replication in vitro, differentially stimulated the activity of both DNA polymerases. RF-A and RF-C independently stimulated DNA polymerase alpha activity 4- to 6-fold, yielding relatively short DNA strands (less than 1 kb) and PCNA had no effect. In contrast, polymerase delta activity was stimulated co-operatively by PCNA, RF-A and RF-C approximately 25- to 30 fold, yielding relatively long DNA strands (up to 4 kb). Neither RF-C nor RF-A appear to correspond to known polymerase stimulatory factors. RF-A was previously shown to be required for initiation of DNA replication at the SV40 origin. Results presented here suggest that it also functions during elongation. The differential effects of these three replication factors on DNA polymerases alpha and delta is consistent with the model that the polymerases function at the replication fork on the lagging and leading strand templates respectively. We further suggest that co-ordinated synthesis of these strands requires dynamic protein-protein interactions between these replication factors and the two DNA polymerases. PMID- 2573522 TI - A mutation in the insulin receptor gene that impairs transport of the receptor to the plasma membrane and causes insulin-resistant diabetes. AB - Insulin binds to a receptor on the cell surface, thereby triggering a biological response within the target cell. Mutations in the insulin receptor gene can render the cell resistant to the biological action of insulin. We have studied a family in which two sisters have a genetic form of insulin-resistant diabetes mellitus. The technique of homozygosity mapping has been used to demonstrate that the mutation causing diabetes in this consanguineous family is genetically linked to the insulin receptor gene. The two insulin-resistant sisters are homozygous for a mutation encoding substitution of valine for phenylalanine at position 382 in the alpha-subunit of the insulin receptor. Transfection of mutant insulin receptor cDNA into NIH3T3 cells demonstrated that the Val382 mutation impaired post-translational processing and retarded transport of the insulin receptor to the plasma membrane. Thus, the mutation causes insulin resistance by decreasing the number of insulin receptors on the surface of the patients' cells. PMID- 2573524 TI - Octamer binding proteins confer transcriptional activity in early mouse embryogenesis. AB - Oct4 and Oct5 are two mouse maternally expressed proteins binding to the octamer motif. Both are found in unfertilized oocytes and embryonic stem cells, whereas Oct4 is also found in primordial germ cells. In this study, the activity of the octamer motif was analysed in two embryonic stem cell lines containing Oct4 and Oct5, the teratocarcinoma-derived cell line F9 and the blastocyst-derived cell line D3. It is known that oligomerization of the octamer motif creates a powerful B-cell specific enhancer. As shown here, this oligomerized transcriptional element is also a very strong enhancer in F9 and D3 embryonic stem cells. After differentiation of the stem cells, both enhancer activity and the amount of the octamer binding proteins decrease. An intact octamer stimulates heterologous promoters in embryonic stem cells, whereas mutations in the octamer motif abolish transcriptional stimulation and binding of the octamer factors. The use of transgenic embryos demonstrates transcriptional activation in the inner cell mass but not in the trophoblast of blastocysts. The results indicate that Oct4 and Oct5 are active early in mouse development. PMID- 2573523 TI - A family of octamer-specific proteins present during mouse embryogenesis: evidence for germline-specific expression of an Oct factor. AB - We have analysed various adult organs and different developmental stages of mouse embryos for the presence of octamer-binding proteins. A variety of new octamer binding proteins were identified in addition to the previously described Oct1 and Oct2. Oct1 is ubiquitously present in murine tissues, in agreement with cell culture data. Although Oct2 has been described as a B-cell-specific protein, similar complexes were also found with extracts from brain, kidney, embryo and sperm. In embryo and brain at least two other proteins, Oct3 and Oct7, are present. A new microextraction procedure allowed the detection of two maternally expressed octamer-binding proteins, Oct4 and Oct5. Both proteins are present in unfertilized oocytes and embryonic stem cells, the latter containing an additional protein, Oct6. Whereas Oct4 was not found in sperm or testis, it is expressed in male and female primordial germ cells. Therefore Oct4 expression is specific for the female germline at later stages of germ cell development. Our results indicate that a family of octamer-binding proteins is present during mouse development and is differentially expressed during early embryogenesis. Protease clipping experiments of Oct4 and Oct1 suggest that both proteins contain similar DNA-binding domains. PMID- 2573525 TI - Novel metabolism of several beta zero-thalassemic beta-globin mRNAs in the erythroid tissues of transgenic mice. AB - Mice that are transgenic for human beta zero-thalassemic beta-globin alleles were generated in order to study how beta zero-thalassemic mutations affect beta globin RNA metabolism in erythroid tissues. Three thalassemic alleles were studied, each of which harbors either a frameshift or a nonsense mutation. These mutations result in the premature termination of beta-globin mRNA translation and an abnormally low level of beta-globin mRNA in the peripheral blood of thalassemic patients. Comparative studies of mice that express any of the beta zero-thalassemic transgenes with mice that express a normal human beta-globin transgene demonstrated that all three thalassemic mRNAs are metabolized in erythroid tissues abnormally. RNA blotting and S1 nuclease transcript mapping revealed for each thalassemic transgene that (i) the full-length mRNA is abnormally short-lived and (ii) in addition to full-length mRNA, three more stable yet smaller RNAs are present. These smaller RNAs are polyadenylated and lack the mRNA 5' end. PMID- 2573526 TI - Homeotic gene expression in the visceral mesoderm of Drosophila embryos. AB - The visceral mesoderm adhering to the midgut constitutes an internal germ layer of the Drosophila embryo that stretches along most of the anteroposterior axis (parasegment 2-13). Most cells of the midgut visceral mesoderm express exclusively one of five homeotic genes. Three of these genes, Antennapedia, Ultrabithorax and abdominal-A are active in parasegmental domains characteristic for this germ layer as they are nonoverlapping and adjacent. The common boundaries between these domains depend on mutual regulatory interactions between the three genes. The same genes function to control gut morphogenesis. Two further homeotic genes Sex combs reduced and Abdominal-B are expressed at both ends of the midgut visceral mesoderm, although absence of their expression does not appear to affect gut morphogenesis. There are no regulatory interactions between these two and the other homeotic genes. As a rule, the anterior limit of each homeotic gene domain in the visceral mesoderm is shifted posteriorly by one parasegment compared to the ectoderm. The domains result from a set of regulatory processes that are distinct from the ones ruling in other germ layers. PMID- 2573528 TI - The ribosomal genes of the mosquito, Aedes aegypti. AB - The characterisation of the ribosomal genes of the mosquito, Aedes aegypti, is described. Preliminary experiments using a cloned Drosophila ribosomal DNA (rDNA) repeat to probe Southern transfers of Ae. aegypti genomic DNA has indicated that the rDNA repeat of Ae. aegypti is 9.0 kb in length and that individual rDNA repeats exhibit a high degree of homogeneity with respect to length and the position of restriction enzyme recognition sites within the rDNA. The preliminary mapping data together with partial digestion experiments demonstrate that, as in all other higher eukaryotes, the rDNA repeats are arranged in a head-to-tail, tandemly repeating manner. The restriction mapping of cloned rDNA repeats confirmed the largely uniform length of the Ae. aegypti rDNA repeat and provided a more detailed physical map of the DNA. A restriction site polymorphism was detected in one clone (Aar9) which contains an extra HincII site, which is not present in three other clones studied (Aar1, Aar3, or Aar7). Transcription mapping has allowed the allocation of identities to the various restriction fragments and the approximate positioning of the transcription unit. The estimate of rDNA repeat copy number in Ae. aegypti (approximately 500 copies per haploid genome) is similar to the estimate reported for the closely related species, Aedes albopictus, of 430 copies per haploid genome. Ribosomal DNA thus comprises approximately 0.6% of the total Ae. aegypti genome. Analysis of the variation of the rDNA repeat unit both within individual mosquitoes and between strains of Ae. aegypti, has severed to confirm the remarkable homogeneity of the rDNA repeat unit in this insect. PMID- 2573527 TI - An essential role of even-skipped for homeotic gene expression in the Drosophila visceral mesoderm. AB - We have analysed homeotic gene expression in the embryonic visceral mesoderm of segmentation mutants by antibody staining against Ultrabithorax, Antennapedia and Sex combs reduced protein. We found that even-skipped (eve) function is crucially required for homeotic gene expression, whereas most other segmentation mutations have only minor effects on position and/or width of the homeotic expression domains in this germ layer. Analysis of pair-rule double mutants indicates that complete loss of homeotic gene activity in the visceral mesoderm, as observed in amorphic eve mutants, correlates with loss of engrailed (en) expression in the epidermis and loss of segmentation. We suggest that the establishment of parasegment borders, a consequence of eve expression and witnessed by subsequent en expression, is a necessary precondition for homeotic gene expression in the visceral mesoderm. PMID- 2573529 TI - Production of human adrenocorticotropin by cleavage of alkaline-phosphatase derived fusion proteins containing repetitive recognition sequences for collagenases. AB - Recombinant plasmids coding for fusion proteins which consist of human adrenocorticotropin joined to N-terminal sequences of Escherichia coli alkaline phosphatase via collagenase-sensitive linkers were constructed and used for the production of these proteins by transformed E. coli cells. It was shown that repetitive linkers of the form -Gly-(Pro-Xaa-Gly)n-Pro- with n greater than or equal to 2 were cleaved by clostridiopeptidase A (Clostridium histolyticum) by orders of magnitude faster than corresponding nonrepetitive sequences (n = 1). The C-terminal cleavage product was Gly-Pro-adrenocorticotropin which could be converted to the authentic hormone by dipeptidyl peptidase IV. On the basis of these enzymatic reactions a procedure for the preparation of pure adrenocorticotropin was developed. Derivatives of alkaline phosphatase containing similar repetitive linker sequences were cleaved by clostridiopeptidase A as efficiently as the adrenocorticotropin fusion proteins. PMID- 2573531 TI - Different large deletions of T cell receptor V beta genes in natural populations of mice. AB - A panel of geographically separate Mus m. domesticus and Swiss mice from several sources was screened for deletions in the T cell receptor variable (V) beta locus. Four out of forty-three strains tested show a deletion identical to or larger than the deletion previously described in SJL mice. To our knowledge, this is the first time that such important V beta deletions are described in inbred or partially inbred wild-derived strains of mice. On the other hand there seems to be very little polymorphism between the remaining V beta genes. Expression of V beta genes in peripheral and intra-thymic T cells was tested using antibodies specific for different V beta polypeptide chains. Flow cytometry analysis revealed a high expression of V beta 6 and V beta 17 genes in the Copacabana Swiss-derived strain and an absence of V beta 17 expression in the WLA wild derived strain. The three Mus m. domesticus strains (WLA, DDO and WBG) having deleted two to three additional V beta subfamilies compared to SJL present no apparent immune deficiencies or autoimmune disorders. We conclude that relatively few V beta genes may suffice for unimpaired survival of wild mice and that there is a selective pressure for the structural conservation of the remaining V beta genes. PMID- 2573530 TI - Conformation of two somatostatin analogues in aqueous solution. Study by NMR methods and circular dichroism. AB - Cyclic-disulfide-containing analogues of somatostatin, Xaa1-Cys2-Xaa3-DTrp4-Lys6 Thr5-Xaa7- Xaa8 [Xaa1 = H or DPhe; Xaa3 = Phe or Tyr; Xaa7 = Cys, Me2Cys or Me2DCys; Xaa8 = OH, Thr8 (OH) or Thr8NH2], were examined in aqueous solution by 1H-NMR spectroscopy and circular dichroism. The influence of the helical nature of the disulfide bridge and the presence of exocyclic residues on biological activity were investigated with particular care. PMID- 2573532 TI - Ifenprodil and SL 82.0715 antagonize N-methyl-D-aspartate (NMDA)-coupled glycine receptor responses in vivo. PMID- 2573533 TI - Continuous and intermittent levodopa differentially affect rotation induced by D 1 and D-2 dopamine agonists. AB - The effects of continuous and intermittent levodopa treatment on rotational behavior induced by dopamine agonists were examined in rats with a unilateral 6 hydroxydopamine lesion of the nigrostriatal dopamine pathway. Chronic administration of levodopa by continuous infusion (90-100 mg/kg per day i.p. by osmotic pump for 19 days with a 3 day washout) enhanced the rotational response to the D-2 dopamine receptor agonist quinpirole, but had no effect on rotation induced by the D-1 agonist SKF 38393 or that due to the non-selective dopamine agonist apomorphine. The rotational responses to the selective dopamine agonists differed dramatically in rats treated with levodopa by intermittent injection (45 50 mg/kg i.p., b.i.d. for 19 days with a 3 day washout): they showed a markedly increased response to quinpirole, a greatly diminished response to SKF 38393, and a modestly enhanced response to apomorphine. Continuous and intermittent treatment resulted in equivalent daily plasma levodopa levels. These findings suggest that the intermittence of central dopamine receptor stimulation may be an important factor in determining the subsequent responses of the dopamine system. The dissociation between the effects of both continuous and intermittent levodopa on D-1 and D-2 agonist-induced rotation indicates that D-1 and D-2 dopamine receptor-mediated mechanisms respond differently to chronic levodopa treatment. PMID- 2573534 TI - Evaluation of selective actions of dopamine D-1 and D-2 receptor agonists and antagonists on opioid antinociception. AB - The effect of the selective dopamine receptor agonists SKF 38393 (D-1) and quinpirole (D-2) on nociception was studied in the mouse tail immersion test. The D-1 receptor agonist induced mild hyperalgesia whereas the D-2 agonist produced antinociception. Pretreatment with either the selective D-1 receptor antagonist SCH 23390 or the D-2 receptor antagonist (-)-sulpiride converted the hyperalgesia produced by the D-1 agonist into an antinociceptive response whereas the effect of the D-2 receptor agonist was significantly antagonised. The antinociceptive response of selective opioid agonists was also studied in combination with selective dopamine receptor agonists and antagonists. Sufentanil (mu-opioid) antinociception was enhanced in animals pretreated with (-)-sulpiride but not SCH 23390. In animals co-administered sufentanil with SKF 38393 there was a reduced antinociceptive effect whilst quinpirole enhanced the action of sufentanil. Likewise, antinociception induced by the kappa-opioid agonist U50,488H was unaltered in animals pretreated with SCH 23390, increased by (-)-sulpiride, and reduced by SKF 38393. delta-Opioid antinociception induced by [D-Ala2,D Leu5]enkephaline remained unmodified following pretreatment with either (-) sulpiride or SCH 23390 but was potentiated in animals which received both the delta-agonist and the D-2 receptor agonist. It is concluded that D-2 receptor agonists not only have intrinsic antinociceptive activity, but can also potentiate opioid-induced antinociception. Similarly, dopamine D-2 receptor antagonists appear to potentiate opioid-induced antinociception in this nociceptive model. PMID- 2573535 TI - Novel alpha 2-adrenoceptor antagonists show selectivity for alpha 2A- and alpha 2B-adrenoceptor subtypes. AB - Pharmacological characterization of mammalian alpha 2-adrenoceptors in various tissues and species has provided evidence for the existence of two alpha 2 adrenoceptor subtypes. These subtypes can be defined in rat and human tissues by prazosin which is alpha 2B selective and oxymetazoline which is alpha 2A selective. In addition to these agents, two types of alpha 2-adrenoceptor antagonists are described which show high affinity and selectivity for the alpha 2A-adrenoceptor and the alpha 2B-adrenoceptor respectively. PMID- 2573536 TI - Intra-hippocampal buspirone in animal models of anxiety. AB - The effect of intra-hippocampal injections of the serotonergic 5-HT1A receptor agonist, buspirone, on rat exploratory activity was evaluated in the 'open field' and 'elevated plus maze' tests. The dose of 2.5 micrograms, but not of 1 microgram, of buspirone administered to the dentate gyrus of the hippocampus increased the time spent on exploration of open arms in the elevated plus maze, as well as it increased the number of central entries in the open field. The results indicate an anti-emotional influence of local stimulation of 5-HT1A receptors by buspirone. PMID- 2573537 TI - In vivo modulation of striatal phosphoproteins by dopaminergic agents. AB - Protein phosphorylation in the brain represents a common target for several second messenger systems. A phosphoprotein (DARPP-32) specifically regulated by cAMP and dopamine has been detected in neurons bearing dopamine D-1 receptors, where it plays a key role in eliciting cAMP-mediated intracellular responses. The endogenous phosphorylation of this cytosolic protein is markedly affected after in vivo acute treatment with the selective D-1 agonist, SKF 38393. The amount of the DARPP-32 dephospho-form measured by a back-phosphorylation assay was decreased by about 30% in agonist-treated animals. This effect was completely counteracted by the concomitant administration of the selective D-1 antagonist, SCH 23390, but not by a selective D-2 antagonist. This first demonstration of in vivo modulation of the phosphorylation state of DARPP-32 could, as a biochemical approach, represent a useful tool to gain further insight into the cascade of biochemical events elicited by specific dopaminergic drugs. PMID- 2573538 TI - Binding of dynorphin A and related peptides to kappa- and mu-opioid receptors: sensitivity to Na+ ions and Gpp(NH)p. AB - We have examined the effects of Na+ ions and 5'-guanylyl imidodiphosphate (Gpp(NH)p) on the equilibrium binding of dynorphin A and of a series of related agonist and antagonist peptides to kappa- and mu-opioid receptors in guinea pig (kappa) and rabbit (mu) cerebellum membrane preparations. The binding to kappa sites of dynorphin A and of the peptides displaying agonist properties was strongly inhibited in the presence of 120 mM NaCl and 50 microM Gpp(NH)p. In contrast, a somewhat lower sensitivity to the inhibitory effect of the two allosteric effectors was observed for the analogues of the series showing antagonist properties. The same general behavior, but more marked, was observed at mu sites, at both mu- and kappa-opioid receptors. The peptides had biochemical properties (binding sensitivity vs. insensitivity to sodium ions and guanine nucleotides) that correlated well with their biological activities (agonist vs. antagonist) previously determined in in vitro pharmacological bioassays. PMID- 2573539 TI - Monoclonal antibodies that bind to hemopoietic stem cells: characterization and immunohistochemical localization of cells expressing gp50-65. AB - The cell surface of hemopoietic stem cells has been shown to express several antigens in common with more mature hemopoietic cells. One set of stem cell antigens is defined by a group of three monoclonal antibodies (13C6, 1C10, and 1A9), selected on the basis of binding to subpopulations of spleen colony-forming stem cells (CFU-S). These antibodies are shown to recognize cell surface glycoproteins of 50-65 kd (gp50-65) that occur widely on hemopoietic cells. Each cell type investigated shows a distinctive pattern of expression of these glycoproteins. To further investigate the presence of gp50-65 on stem cells, low density bone marrow cells were labeled with 1C10 and sorted according to fluorescence intensity. Most (73%) of the stem cells (CFU-S10) were recovered in the two most highly fluorescent fractions containing 2.6% of starting marrow cells. Immunohistochemistry of frozen sections of normal spleen and spleens during repopulation after lethal irradiation and bone marrow transplantation showed that the most strongly 1C10-labeled cells occurred under the splenic capsule and along trabeculae. Although many of these cells were also alpha naphthylacetate esterase positive and Mac-1 positive, indicating cells of the monocyte-macrophage lineage, a distinctive population of singular cells were stained with 1C10 alone. These cells were negative for surface Ig and closely corresponded with a small population of cells in a similar location that were doubly labeled with 1C10 and anti-Thy-1. These results show that stem cells express high levels of gp50-65 and suggest that stem cells can be identified by immunohistochemical methods using dual labeling procedures. PMID- 2573540 TI - Immunological and molecular biological identification of a true case of T-hairy cell leukaemia. AB - A hairy cell leukaemia (HCL) patient is presented in whom the peripheral blood mononuclear cells (PBMCs) carried suppressor T-cell markers (CD3+, CD2+, CD8+/CD4 , CD38+). Analysis of genomic DNA of PBMNC showed the presence of a monoclonal population of T cells, the T-cell receptor (TCR) beta-chain gene being rearranged on both alleles (DR/DR), while the immunoglobulin (Ig) heavy chain-genes were in germline configuration. The neoplastic cells were found to react with the monoclonal antibody RAB-1 - originally described as belonging to the B lineage restricted monoclonal antibodies - and to carry RAB-1/CD-8 in a double marker assay. Natural killer activity of PBMNCs against K562 target cells was severely reduced, while the cells were found to exert strong antibody-dependent cellular cytotoxicity. PMID- 2573541 TI - An antiserum to the extracellular domain of the Alzheimer amyloid precursor recognizes 70 and 88 kDa brain proteins. AB - An antiserum raised to the extracellular domain (residues 556-566) of the Alzheimer amyloid precursor protein recognized 70 and 88 kDa proteins in Western blots of rat, Alzheimer, Down's syndrome and control human brain separated by SDS PAGE. The 70 kDa protein band was resolved into 5 spots by two-dimensional electrophoresis. The findings provide further evidence that a 70 kDa protein is a metabolite of the amyloid precursor protein and reveal an 88 kDa protein which was reduced in 3 out of 6 brains with Alzheimer pathology. PMID- 2573543 TI - Putative N-terminal splitting enzyme of amyloid A4 peptides is the multicatalytic proteinase, ingensin, which is widely distributed in mammalian cells. AB - The main characteristic changes observed in Alzheimer's disease (AD) are the presence of neurofibrillary tangles and the deposition of amyloid A4 peptides. The most abundant amyloid A4 peptide species in AD (which we tentatively named A4') is composed of 39 amino acids, which is devoid of the 3 N-terminal amino acids, Asp-Ala-Glu, of the originally reported A4 peptide. We synthesized a model peptide substrate, Suc-Ala-Glu-methylcoumarinamide (MCA), to identify the proteinase that splits the A4' peptide. DEAE-cellulose column chromatography of rat liver and porcine brain extracts showed that only one peak material digested the synthetic substrate at pH 8. The results for the final preparation indicate that the Suc-Ala-Glu-MCA-degrading enzyme is a high-molecular-mass proteinase, with a molecular mass of above 500,000, and is composed of several low-molecular mass subunits. These results suggest that a non-lysosomal multicatalytic proteinase (we named this enzyme ingensin (ingens = large in Latin). Ishiura, S. et al. (1985) FEBS Lett. 189, 119-123) catalyzes the above reaction. Antiserum against the purified multicatalytic proteinase, ingensin, crossreacted with the purified Suc-Ala-Glu-MCA-degrading proteinase. It is likely that ingensin shows a similar action toward amyloid precursor protein (APP) in vivo. PMID- 2573542 TI - Acidic and basic fibroblast growth factors similarly regulate the rate of biosynthesis of rat astroblast proteins. AB - Quiescent rat astroblasts in culture have been treated for various periods of time with acidic and basic fibroblast growth factors. Both factors elicited similar effects on the cell proliferation and glutamine synthetase activity. The rate of biosynthesis of the proteins analyzed on autoradiograms of polyacrylamide gels after two-dimensional electrophoresis was also similarly modulated by the two growth factors. These results suggest that the two fibroblast growth factors act through the same membrane receptors on rat astroblasts in culture. PMID- 2573545 TI - [Deontological and psychological bases for obtaining the anamnesis in pediatrics]. PMID- 2573544 TI - P-glycoproteins encoded by mdr 1b in murine gravid uterus and multidrug resistant tumor cell lines are differentially glycosylated. AB - There are 3 members of the multidrug-resistance gene family expressed in mouse. Only one of these, mdr 1b, and its gene product P-glycoprotein are induced to high levels in the mouse endometrium during pregnancy. It is shown here that P glycoprotein in the gravid uterus is significantly larger (Mr 155,000) compared to P-glycoprotein encoded by mdr 1b in a murine multidrug-resistant cell line (Mr 140,000). However, both species co-migrate after enzymatic removal of N-linked sugars (Mr 125,000). These results demonstrate that differential glycosylation of the mdr 1b gene product contributes to molecular heterogeneity found in P glycoprotein from normal and multidrug-resistant cells. PMID- 2573546 TI - Dehydroepiandrosterone and related steroids inhibit mitochondrial respiration in vitro. AB - 1. Dehydroepiandrosterone (DHEA) inhibited mitochondrial respiratory rates in the presence of tricarboxylic acid cycle intermediates with the exception of succinate. 2. Several other steroids tested for inhibitory activity on mitochondrial state 3 rate showed inhibitory potencies ranging from 0-90%. 3. Mitochondria isolated from adrenals, heart, kidneys, brain and brown adipose tissue were also inhibited by DHEA. 4. The aspartate malate shuttle system was inhibited, but the alpha-glycerophosphate shuttle was totally insensitive to DHEA. 5. The high-amplitude swelling of mitochondria in hypotonic media was inhibited by DHEA and a few other steroids. PMID- 2573547 TI - Formation of ketone bodies by resting lymphocytes. AB - 1. Both beta-hydroxy-beta-methylglutaryl-coenzyme A synthase and lyase activities are present in rat mesenteric lymphocytes: all of the synthase and almost all (80%) of the lyase were present in the mitochondrial compartment of the cell. 2. A high rate of acetoacetate formation was observed in mesenteric lymphocytes incubated in vitro for 60 min in the absence of added substrate; addition of pyruvate or glutamine increased the "endogenous" rate of acetoacetate formation by about 30%. 3. The rates of ketone body formation are similar to maximal rates observed for rat liver. 4. It is suggested that the high rate of endogenous acetoacetate production occurs from long chain fatty acids: this suggestion is consistent with the reported high "endogenous" rate of O2 consumption by lymphocytes. 5. Of the pyruvate metabolized via pyruvate dehydrogenase in lymphocytes, ca 50-70% could be accounted for as acetoacetate, acetate, 3 hydroxybutyrate and citrate: the fate of the remainder is not known. 6. There was a high rate of endogenous acetoacetate formation by isolated mitochondria from these cells. 7. The rate was doubled by addition of pyruvate or butyrate; it was trebled by addition of propionate, ADP or carbonyl cyanide trichloro methoxyphenylhydrazone; but it was decreased by addition of antimycin A or glutamine. 8. It is suggested that the high rates of acetoacetate formation in these cells acts as a dynamic "buffer" system for the acetyl coenzyme A (CoA) concentration: that is, acetyl CoA is always available for fatty acid synthesis, cholesterologenesis, chain extension of fatty acids or acetylation of proteins (e.g. for covalent control of their activity) which will be demanded at different stages of the cell cycle. 9. This is another example of branch-point sensitivity in control in cells with the potential for rapid cell division. PMID- 2573548 TI - Uptake and stimulation-evoked release of tritiated neurotransmitters from slices of the rat rostral ventral medulla containing the nucleus raphe magnus: implications for pathways controlling nociception. AB - 1. The rostral ventral medulla plays a central role in the integration of nociceptive control. 2. Slices of this area of the brainstem may be labelled with tritiated noradrenaline, dopamine, serotonin, GABA and choline. 3. Uptake was greatest for noradrenaline and dopamine, GABA was intermediate and serotonin and choline were poorly accumulated. 4. Conditions for the release of all transmitter candidates except acetylcholine were established using either potassium or electrical stimulation and release was proven to be calcium dependent. 5. Electrophysiological and microinjection data are at variance with the commonly assumed actions of noradrenaline and dopamine and can be rationalized by the presence of a GABA interneuron integrating nociceptive input to the nucleus raphe magnus. PMID- 2573549 TI - Estradiol attenuates prolactin secretion and phosphoinositide hydrolysis in MMQ cells. AB - We previously isolated a clonal cell line, designated MMQ, which only secretes prolactin (PRL) and whose secretory process is nonresponsive to thyrotropin releasing hormone (TRH) and angiotensin II (AII). In the present study, we injected MMQ cells into rats to determine whether the tumor cells would become responsive to secretagogues when subsequently propagated in vitro. We also investigated what effects in vivo administration of 17 beta-estradiol would have on secretagogue-induced PRL release and on intracellular biochemical mechanisms in these cells. MMQ cells were implanted subcutaneously in the backs of female rats. One group was injected with 100 micrograms polyestradiol phosphate (PEP) every 5 days, a second with saline. The inoculants grew into solid tumors within 3 weeks. The day after the tumors were removed and enzymatically dispersed, the cells, now designated MMQt cells, were perifused in vitro. Basal PRL released by MMQt cells was approximately 1 ng/min/10(7) cells and perifusions with 100 nM TRH or AII for 5 min significantly increased PRL release above baseline (integrated areas: 1.8 +/- 0.4 and 5.2 +/- 1.3 ng/10(7) cell, respectively; P less than 0.01). Two ng/ml maitotoxin (MTX), a calcium channel activator, increased PRL release (38.2 +/- 6.7 ng/10(7) cells; P less than 0.01). In PEP-treated perifused MMQt cells, basal in vitro PRL release was not different from that observed in the control group, but the responses to TRH, AII and MTX were greatly attenuated (TRH: 0.6 +/- 0.1, AII: 1.3 +/- 0.2 and MTX: 9.2 +/- 2.5 ng/10(7) cells).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573550 TI - The survival of chick retinal ganglion cells in response to brain-derived neurotrophic factor depends on their embryonic age. AB - The survival effects of brain-derived neurotrophic factor (BDNF) on the ganglion cells of the chick retina were studied in vitro at different embryonic ages. We found these effects to be strongly age-dependent: at E5, when the first ganglion cell axons have crossed the optic chiasm, but not yet reached the tectum, ganglion cells survived on a laminin substrate irrespective of the presence or absence of BDNF. At E6, when the axons of the first-generated ganglion cells reached the rostral pole of the tectum, the ganglion cells began to show a dependency on BDNF for survival, but the majority of them were alive after 2 days in vitro in the absence of BDNF. With increasing age, the BDNF dependency for survival increased, and at E11, the majority of the ganglion cells plated were dependent on BDNF for survival. It is at this age that the maximal number of axons can be found in vivo in the optic nerve, the subsequent elimination of ganglion cells and their axons resulting in the loss of hundreds of thousands of them over the next few days. Taken together, these data indicate that retinal ganglion cells depend on BDNF for survival only when their axons have reached their target in vivo. This situation is reminiscent of that described in the peripheral nervous system for the nerve growth factor responsiveness of mouse trigeminal sensory neurons during the period of innervation of their target. PMID- 2573551 TI - Inhibition of the adrenergic phenotype in cultured neural crest cells by norepinephrine uptake inhibitors. AB - Tricyclic antidepressants in combination with in vitro clonal analysis of quail neural crest cells were used to examine the role the norepinephrine uptake mechanism might play in the development of adrenergic neural crest derivatives. Norepinephrine (NE) uptake inhibitors blocked expression of the adrenergic phenotype by neural crest cells. The degree of inhibition of phenotypic expression correlated with the potency and specificity of the uptake inhibitors. The drugs acted during the early phase of in vitro development, i.e., several days before overt expression of the adrenergic phenotype in clonal culture. They were nontoxic, and a chronic exposure of the cells to NE uptake inhibitors was necessary to cause an effect. These observations suggest that norepinephrine and possibly related neurotransmitters play a direct or indirect role in the expression of the adrenergic phenotype by neural crest cells and that tricyclic antidepressants may affect neurogenesis during sensitive stages of embryonic development. The data may reflect in vivo mechanisms, since there are neurotransmitters present in the migratory pathway of presumptive sympathetic neurons and the norepinephrine uptake system is expressed in the embryo by these cells before they synthesize and accumulate catecholamines. PMID- 2573552 TI - The determination of the adrenal medullary cell fate during embryogenesis. AB - One subset of neural crest cells, the sympathoadrenal precursors, undergoes a switch in phenotype expression, when they invade the adrenal anlagen and become associated with adrenocortical cells. To investigate the mechanisms responsible for the conversion of noradrenaline synthesizing precursors to adrenaline producing endocrine chromaffin cells we studied the role of glucocorticoids on the initial induction of adrenaline synthesis in embryonic adrenals and cultures of highly purified chromaffin precursor cells. We could show that in vivo differentiation of rat chromaffin precursors commences between 16.3 and 17.3 days of gestation. While adrenaline and the activity of the enzyme phenylethanolamine N-methyltransferase (PNMT), which converts noradrenaline to adrenaline, were present at Embryonic Day 17.3 (E17.3), they were not detectable in E16.3 adrenals. Small amounts of corticosterone were present in E16.3 adrenals and plasma, but in parallel with the initial induction of adrenaline biosynthesis, a sharp rise in organ and plasma glucocorticoid levels occurred until E17.3. Chromaffin precursor cells, isolated at E16.3 and cultured for 4 days, failed to express PNMT activity and adrenaline. However, 0.1 nM dexamethasone was already sufficient for the initial induction of adrenaline and its synthesizing enzyme. Specific glucocorticoid binding of freshly isolated chromaffin (precursor) cells revealed a developmental increase during embryogenesis, yet no glucocorticoid binding sites were detectable in chromaffin precursor cells at E16.3. They appeared at E17.3 in parallel with the initial induction of adrenaline biosynthesis and the enormous rise of adrenal and plasma corticosterone levels. We therefore conclude that glucocorticoids are essential and sufficient to trigger the differentiation of noradrenergic sympathoadrenal precursors to adrenergic chromaffin cells after a functional glucocorticoid receptor system has been established. PMID- 2573553 TI - Effects of glucagonlike peptide I-(7-36) on release of insulin, glucagon, and somatostatin by rat pancreatic islet cell monolayer cultures. AB - Glucagonlike peptide I (GLP-I-(7-36] is cleaved from proglucagon in ileal epithelial cells and increases in human plasma after nutrient ingestion. This peptide has been shown to stimulate insulin secretion in vitro and in vivo and thus potentially acts as an incretin. To characterize its action on islet cells, the release of insulin, glucagon, and somatostatin by rat pancreatic islet monolayer cultures at varying concentrations of GLP-I-(7-36) was measured. The interaction of GLP-I-(7-36) with nutrient substrates was assessed by adding amino acids and differing glucose concentrations to the cultures. Islet cell cultures (n = 5) were incubated for 1 h in medium containing 1.67 or 16.7 mM glucose or 1.67 mM glucose supplemented with amino acids and GLP-I-(7-36) at 10(-13)-10(-7) M. Hormone release was compared with control cultures containing no GLP-I-(7-36); 1.67-16.7 mM glucose with and without GLP-I-(7-36) at 10(-11) M; and 1.67, 3.3, 8.3, or 11.1 mM glucose alone or supplemented with amino acids, GLP-I-(7-36) 10( 11) M, or both amino acids and GLP-I-(7-36). In medium with 1.67 or 16.7 mM glucose or 1.67 mM glucose and amino acids, GLP-I-(7-36) increased insulin secretion two- to threefold over control at concentrations of 10(-9), 10(-11), and 10(-12) M, respectively. In medium with increasing concentrations of glucose, GLP-I-(7-36) at 10(-11) M significantly increased insulin secretion at glucose concentrations greater than or equal to 3.34 mM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573554 TI - Insulin regulation of lipolysis in nondiabetic and IDDM subjects. AB - To determine whether insulin regulation of lipolysis is abnormal in subjects with poorly controlled insulin-dependent diabetes mellitus (IDDM), free-fatty acid flux ([1-14C]palmitate) was measured under conditions ranging from complete insulin withdrawal to hyperinsulinemia. Seven nondiabetic and seven IDDM subjects were studied with the pancreatic clamp technique to control plasma insulin, growth hormone, and glucagon concentrations at the desired levels. Preliminary studies were performed to validate the experimental design. The palmitate flux response to insulin withdrawal (2.5 +/- 0.2 vs. 2.5 +/- 0.2 mumol.kg-1.min-1) and maximally antilipolytic insulin concentrations (0.17 +/- 0.02 vs. 0.23 +/- 0.03 mumol.kg-1.min-1) were not different in nondiabetic and IDDM subjects, respectively. In contrast, IDDM subjects required significantly greater plasma free-insulin concentrations to result in equivalent suppression of palmitate flux compared with nondiabetic subjects. Lipolysis was found to be very sensitive to insulin in nondiabetic humans, with half-maximal suppression occurring at plasma free-insulin concentrations of approximately 12 pM (less than 2 microU/ml). We conclude that adipose tissue lipolysis is normally exquisitely sensitive to insulin and that sensitivity, but not responsiveness to insulin, is impaired in poorly controlled IDDM. PMID- 2573555 TI - Insulin prevention of altered muscarinic receptor-G protein coupling in diabetic rat atria. AB - Right atria from rats rendered diabetic by injection of streptozocin (STZ-D) for 8-10 wk are supersensitive to the negative chronotropic effects of muscarinic agonists but have decreased levels of muscarinic receptors and acetylcholinesterase activity. Insulin treatment completely prevents the development of these changes. The proportion of atrial muscarinic receptors displaying high-affinity agonist binding is lower in STZ-D rats; however, the sensitivity of high-affinity agonist binding to regulation by a guanine nucleotide (5'-guanylylimidodiphosphate) is greater in atria from diabetic rats. Again, insulin treatment eliminates these differences. These findings indicate that alterations in atrial muscarinic systems in STZ-D rats are a consequence of the elaboration of the diabetic state and suggest that an alteration of functional muscarinic receptor-G protein coupling contributes to the altered physiological responsiveness of the heart in diabetes. PMID- 2573557 TI - Inheritance of restriction fragment length polymorphisms in Agaricus brunnescens. AB - The cultivated mushroom, Agaricus brunnescens, is secondarily homothallic; most basidia produce only two basidiospores, each of which receives two of the four post meiotic nuclei. The segregation of restriction fragment length polymorphisms (RFLPs) detected by four plasmid probes carrying single-copy nuclear DNA of Agaricus was followed in seven parental strains including commercial, wild collected, and artificially synthesized heterokaryons. Of a total of 367 single spore progeny examined, 351 (95.6%) were heteroallelic at all RFLP loci heteroallelic in the respective parents. Of the 16 segregant isolates, ten (2.7% of the total) were homoallelic at all segregating loci assayed, suggesting that these isolates were most probably derived from rare spores that had received only a single postmeiotic nucleus. Some of these ten isolates had recombinant genotypes. Only five isolates (1.4% of the total) showed homoallelism at one of the loci heteroallelic in the parent, while remaining heteroallelic at other loci. These five genotypes suggest that a crossover had occurred between a marker locus and its respective centromere. Taken together, the results suggest that meiosis in A. brunnescens is accompanied by low levels of recombination and that nonsister nuclei are preferentially incorporated into basidiospores after meiosis II. PMID- 2573556 TI - Association of IDDM and attenuated response of 2',5'-oligoadenylate synthetase to yellow fever vaccine. AB - Basal and yellow fever vaccination-induced 2',5'-oligoadenylate synthetase (2',5'A) activity was determined in blood mononuclear cells (peripheral blood lymphocytes [PBLs]) from insulin-dependent diabetes mellitus (IDDM) and matched control subjects. The live attenuated yellow fever vaccine represented a primary stimulus in all subjects. First, basal 2',5'A activity increased severalfold in response to yellow fever vaccination. In IDDM subjects, this increase was significantly lower (P = .025). Second, the 2',5'A activity increased proportionately to the higher basal 2',5'A activity in IDDM subjects. In control subjects, the increase in 2',5'A activity was not dependent on the basal activity. There was no relationship between basal or stimulated 2',5'A activity and age, sex, duration of IDDM, age at onset of IDDM, metabolic control, or HLA DQ beta-chain gene polymorphism. There is a direct relationship between 2',5'A activity and latent viral infections associated with the presence of double stranded RNA and with cellular interferons (IFNs) formed in response to viral infections. The higher basal 2',5'A activity (P = .05) in relation to the stimulated activity may therefore signify a latent infection or the presence of double-stranded RNA in PBLs of IDDM subjects. In vitro stimulation of PBLs showed increased IFN sensitivity in IDDM subjects. Analysis of 2',5'A activity is proposed to be a sensitive measure of the activation of the IFN system and the level of latent infectivity. PMID- 2573559 TI - [Study of polymorphism in human rRNA gene clusters. Population and familial analysis of basic types of RFLP]. AB - The frequency and average amount of copy number per genome were defined for standard and a number of new variants of BamHI 5'-NTS RFLP from populations of Moscow, Riga and individuals with Down syndrome. It was demonstrated that the populations studied differ neither in population frequency nor in the average amount of copy number of the variants. New variants were detected in the EcoRI 3' NTS RFLP system and their amplification, as well as discordance among MZ twins. Possible target for methylation in the HindII site of 3' end of 28S rRNA gene was revealed. Analysis of data obtained demonstrated inefficiency of using the RFLP systems in systematic mapping of NOR-chromosomes. Our data also suggested a possible role of amplification of one copy repeated unit rRNA genes in their evolution. PMID- 2573560 TI - Regulation of chicken alpha and beta actin genes and their hybrids inserted into myogenic mouse cells. AB - We have investigated the regulation of intact non-muscle (beta) and muscle specific (skeletal alpha) chicken actin genes and of hybrids of these two genes (alpha 5'-beta 3' and beta 5'-alpha 3') transferred into the mouse myogenic non fusing cell line BC3H1. BC3H1 cells express members of the actin multigene family in a differentiation-dependent manner. When proliferating, the cells accumulate large amounts of non-muscle actin mRNA; when the cells are induced to differentiate, the amount of non-muscle actin mRNA decreases and the amount of muscle-specific actin mRNA increases. The transferred beta-actin gene is efficiently expressed in undifferentiated cells and appropriately down-regulated upon differentiation. In contrast, the transferred alpha-actin gene is inefficiently expressed and not consistently up-regulated. Results with the intact and hybrid genes, taken together, are consistent with the hypothesis that both 5' and 3' halves of these genes contain sequences important in regulating the efficiency and/or developmental timing of their expression in BC3H1 cells. By nuclear run-on experiments we found no evidence for gene-specific changes in the rate of transcription of the transferred actin genes during myogenesis. We conclude that the differentiation-dependent changes in expression of the intact beta-actin gene in BC3H1 cells must be regulated at the post-transcriptional level. PMID- 2573558 TI - Haplotypes that are mosaic for wild-type and t complex-specific alleles in wild mice. AB - Two outstanding problems pertaining to the population dynamics and evolution of the t complex in mice concern the frequency of t haplotypes in the wild and the degree to which these haplotypes recombine with their wild-type homologs. To address these problems, the frequency and distribution of several t complex associated restriction fragment variants in wild mice were estimated. Sixty-four versions of chromosome 17 from wild-derived Mus musculus musculus and Mus musculus domesticus were examined with DNA probes for six loci within the t complex that exhibit restriction fragment variation. All six probes detect variants that have heretofore been found exclusively associated with the t complex. Haplotype analysis of wild-derived chromosomes revealed a high frequency (45.3%) of "mosaic" haplotypes with a mixture of t-specific and wild-type variants and only one haplotype with t-specific variants at all six loci. When 12 well-characterized t haplotypes isolated from diverse geographic regions were analyzed, only three had a complete set of t-specific restriction fragments for the six loci examined. The preponderance of mosaic haplotypes in both groups of mice can be explained by any one of the following hypotheses: genetic recombination between t haplotypes and their wild-type homologs, the persistence in wild populations of haplotypes that have descended from ancestral partial t haplotypes, or that the restriction fragment variants fixed in the ancestral t haplotype were also fixed in some wild-type haplotypes. There is evidence to support all three of these hypotheses in our data. The allelic composition of some mosaic haplotypes indicates that they may have been formed by segmental recombination, either double crossing over or gene conversion, rather than by simple single crossovers. The occurrence of indistinguishable mosaic haplotypes in both M. m. musculus and M. m. domesticus suggests that these haplotypes are ancestral rather than recently derived. PMID- 2573561 TI - A mouse thymidylate synthase pseudogene derived from an aberrantly processed RNA molecule. AB - A DNA fragment containing a mouse-thymidylate-synthase(TS) processed pseudogene was cloned and analyzed. Comparison with the sequences of the mouse TS-encoding gene (ts) and cDNA revealed that the pseudogene started at one of the normal 5' termini of TS mRNA, ended with a poly(A) tail, and was flanked by 16-nucleotide (nt) direct repeats. The region corresponding to the open reading frame was 97.3% identical to that of the cDNA. Two unusual features were observed. First, the poly(A) tail of the pseudogene was located 2 kb downstream from the normal location. Second, the final 10 nt of intron 5 were retained in the 'coding region' of the pseudogene. Therefore, it appears that the pseudogene was derived from a nonfunctional TS 'mRNA' that was aberrantly spliced and polyadenylated. Analysis of the sequence of intron 5 of the ts gene revealed the presence of an alternative 3' splice site 10 nt upstream from the normal splice site. S1 nuclease protection assays showed that about 10% of TS mRNA isolated from mouse cells was spliced at the alternative site. PMID- 2573562 TI - Characterization of different forms of rat mammary gland acetyl-coenzyme A carboxylase mRNA: analysis of heterogeneity in the 5' end. AB - Acetyl-coenzyme A carboxylase (ACC; EC 6.4.1.2) catalyzes the rate-limiting reaction in the biogenesis of long-chain fatty acids. We have previously reported the coding sequence of ACC mRNA from the mammary gland of the lactating rat. The existence, in this tissue, of several forms of ACC mRNA with different 5' untranslated regions has now been established. Two mRNAs constitute the major ACC mRNA species, they differ from one another in the presence or absence of a 61 nucleotide fragment at the center of the 5'-untranslated region. Multiple forms of ACC mRNA might originate through differential splicing of the primary transcript. PMID- 2573563 TI - The effects of a program for faculty development in geriatrics for physician assistant teachers. AB - A Geriatric Education for Physician Assistant (PA) Faculty program enrolled teachers from 21 of the 55 PA training programs in the country to participate in three-week training sessions at Stanford University. The faculty trainees took part in lecture/discussion, audio-visual review, clinical training, and individual conferences. They prepared a lecture and a complete geriatric teaching plan for their home program, and were assigned a text and numerous articles to read. Measures of effect of the training found the following: a significant increase in knowledge scores, although the trainees came into the program with relatively high scores; a heightened awareness and increased positive attitudes toward aging; high ratings of performance on a functionally oriented comprehensive health assessment; and augmented geriatric curriculum and clinical training in their home PA programs. PMID- 2573564 TI - Physiological targets of superoxide anion and hydrogen peroxide in reperfusion injury. AB - Current dogma associates reperfusion injury with the introduction of reactive oxygen species (ROS) into the ischemic tissue. The sources of ROS under discussion are xanthine oxidase in the endothelium of small vessels and/or invaded polymorphonuclear leukocytes (PMN). The beneficial effects of both superoxide dismutase and catalase suggest an involvement of superoxide anions and hydrogen peroxide in this pathophysiological process, without describing the targets of their action. In our work we demonstrate that these two ROS effectively interact with two enzymes. Superoxide anions inhibit soluble guanylate cyclase. Its product, cGMP, is considered to antagonize platelet activation and to cause smooth muscle relaxation. Thus O2- can intensify platelet aggregability and small vessel occlusion. Similar effects are elicited by H2O2, which shifts the dose response curve of several agonists towards smaller concentrations by activating cyclooxygenase. This enzyme provides the substrate for thromboxane synthase which generates TxA2, the most potent physiologically occurring platelet aggregating and smooth muscle contacting agonist. These results lead us to the suggestion that the influence of the oxidative burst of PMN in the phenomenon of reperfusion injury should be reconsidered. PMID- 2573565 TI - Histamine-2-receptor antagonists in gastro-oesophageal reflux. PMID- 2573566 TI - New insights into the mechanisms which regulate IgE synthesis in man. AB - The studies summarized here provide further insight into the cellular and molecular mechanisms involved in the regulation of human IgE synthesis. They clearly demonstrate that IL-4 is the essential factor for induction of human IgE synthesis, since no substantial in vitro IgE production can be obtained in the absence of this lymphokine. Another T cell-derived lymphokine, IFN-gamma, negatively regulates the IgE synthesis induced by IL-4. These two lymphokines can be produced by different T helper cells, but they can also represent the product of the same T cell clone (TCC). In this case, the possibility that a given TCC provides helper function for IgE seems to be dependent upon the balance between the amounts of the two lymphokines produced. Additional cellular and/or molecular signals are involved in IL-4-dependent IgE synthesis. First of all, a direct T-B interaction is required to precede the activity of IL-4. This interaction does not necessarily consist of cognate interaction between T and B cells, as occurs for IgE synthesis induced by alloreactive TCC. In the presence of exogenous IL-4 virtually all CD4+, and even a number of CD8+, TCC can provide the signaling required by B cells to synthesize IgE. An interaction between some adhesion molecule, more expressed on activated than resting T cells, and its receptor on B cells is sufficient to prepare resting B cells to synthesize IgE in response to IL-4. Furthermore, T cells also contribute to IL-4-dependent IgE synthesis by releasing IL-2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573567 TI - Cytochemistry of dipeptidylaminopeptidase IV in T lymphoblastic leukaemia lymphoma. AB - The cytochemical distribution of dipeptidylaminopeptidase IV (DAP IV) was studied in 5 cases of T lymphoblastic leukaemic lymphoma and 12 cases of acute T lymphoblastic leukaemia, in order to ascertain differences between the enzyme positivity patterns of T cells at different stages of differentiation. Early thymic phenotype cases were almost completely negative; those of intermediate and mature thymic phenotype showed positivity in various percentages of blasts: either a single coarse granule or many coarse and small granules were detected. In mature phenotype cells a particularly intense DAP IV reaction was observed. In conclusion, our findings suggest that DAP IV reaction could be a useful tool for the cytochemical characterization of T acute leukaemia subtypes. PMID- 2573568 TI - [Can beta blockers reduce cardiac morbidity and mortality in hypertension]. PMID- 2573569 TI - Isolation and analysis of DNA marker revealing restriction fragment length polymorphism from X chromosome specific DNA library. AB - A human X chromosome specific DNA library was constructed from flow sorted metaphase X chromosomes. Twenty eight single-copy containing phage clones from this X chromosome library were tested for polymorphism against a panel of DNAs from several unrelated individuals, digested with eight restriction enzymes. One (named lambda 33) of these phage clones revealed high frequency two allele polymorphism with the restriction enzyme Msp I and was regionally mapped to the chromosome Xpter by two methods, including Southern analysis with a mapping panel of cell hybrids and quantitative hybridization. The polymorphism appears to be the result of base pair substitutions or modifications rather than DNA rearrangements. In order to examine the heritability of two alleles, the DNAs from four members of a family spanning three generations were examined. The alleles are consistent with their inheritance as a classic X-linked Mendelian locus. Probe lambda 33 will serve as a marker for linkage studies with known polymorphic loci as well as to establish linkage with X-linked diseases. PMID- 2573570 TI - [Genetic analysis of the class II region of rat major histocompatibility complex (RT1)]. AB - Class II subregions of rat major histocompatibility complex (RT1) consist of RT1. B (HLA-DQ equivalent), D (HLA-DR equivalent), and a newly defined H (HLA-DP equivalent) subregions, arranged in the order of RT1.A (class I-H-B beta (beta 1, beta 2)-D (alpha, beta) RFLP analyses of RT1. B and D subregions confirmed that RFLP patterns were correlated well with the allelic specificities which have been determined by serology or MLR. RT1.B locus of BDIX was shown to have RT1.Bd. RFLF analyses of intra-RT1 recombinant rats revealed that recombinations occurred at the sites between RT1.H and B in NIG III, WIN, KGH, between RT1.A and H in DA. BI, BDIX, and between RT1. H alpha and H beta in LEJ. Taken together, the order of genes around H subregion was determined as follows: RT1.A (class I)-H beta-H alpha-B-. Expressions of H subregion genes has not been known. However, Northern hybridization analysis with DP alpha probe detected mRNA of approximately 1.7 kb, suggesting the transcription of RT1. H alpha gene. PMID- 2573571 TI - [Molecular-pathology analysis of regulatory region and transformation in JC virus]. AB - JC virus, an agent that causes a human demyelinating disease, progressive multifocal leukoencephalopathy (PML), is known to induce brain tumors in hamsters. The regulatory region of the virus reported to be implicated in its tissue specificity and to be highly variable among different isolates. In a study of the regulatory region, first we isolated the JC virus genome directly from the brain of a patient with suspected PML. We amplified the regulatory region of the virus by using the polymerase chain reaction, and we compared with three JC viruses which had been isolated previously (MAD-1, MAD-2, and TKY-1). The brain tissue was shown to be infected with JC virus, as confirmed by the presence of viral antigen on immunohistochemical testing and by the demonstration of viral nucleic acids upon in situ hybridization. Southern blot analysis of the regulatory region disclosed that the isolate from our patient, which was designated as NYS-1, was roughly homologous in all three types of JC viruses, with preservation of the constant regions. However, the cut site for PvuII in NYS 1 was different from that in MAD-1 and MAD-2, and a pair of cut sites, for Nco I PvuII, was unique to TKY-1 and absent in NYS-1; this showed that NYS-1 was a new type of JC virus. Second, we compared the regulatory regions of TKY-1 and its viral DNA integrated in a hamster brain tumor cell line (I-23) induced by TKY-1. We found no great difference between these two genomes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573573 TI - Characterization of alpha-adrenoceptors involved in central thermoregulation in rabbits. AB - Intracerebroventricular (icv) injection of methyldopa induced body temperature changes in the rabbits. The dose of 100 micrograms/kg did not produce any significant change on body temperature whereas 250 micrograms/kg of the drug induced hyperthermia. Higher dose of 500 micrograms/kg produced initial hypothermia which was followed by hyperthermia. On further increase of the dose to 1 mg/kg, consistent hypothermia was evident. Prazosin, a specific post synaptic alpha 1 adrenoceptor blocker, induced hypothermia whereas piperoxan (presynaptic alpha 2 antagonist) produced hyperthermia. The pretreatment with prazosin, blocked the hyperthermic response of methyldopa. The initial hypothermia by 500 micrograms/kg of methyldopa was also potentiated. The pretreatment with piperoxan completely blocked the hypothermia but had no effect on hyperthermic response of methyldopa. Pretreatment of rabbits with both prazosin and piperoxan completely blocked the hypothermia as well as hyperthermic response of methyldopa. Thus it appeared that both presynaptic alpha 2 and postsynaptic alpha 1 adrenoceptors are involved in central thermoregulation in rabbits. PMID- 2573572 TI - A multicenter, randomized, double-blind trial of somatostatin in the management of acute hemorrhage from esophageal varices. AB - A prospective, randomized, placebo-controlled, double-blind, multicenter clinical trial of intravenous somatostatin (Stilamin; Serono Laboratories, Inc., Randolph, MA) was performed in 102 patients with actively bleeding esophageal varices from August, 1985, to November, 1986. Patients had major hemorrhage indicated by hematemesis or melena and evidence of significant blood loss. For entry, patients had to have endoscopic demonstration of active bleeding from esophageal varices or stigmata of recent hemorrhage and bright red blood in the gastric aspirate with no other source of bleeding found. Randomized patients received identical appearing somatostatin or placebo for a 30-hr study period. Those given somatostatin received a 250-micrograms bolus and a 250-micrograms per hr infusion with repeat bolus and doubling of the infusion if the bleeding was not controlled. In retrospect, 18 patients could not be evaluated. Of the 84 evaluable patients, 48 received somatostatin and 36 placebo. They were comparable in age, gender, severity of liver disease and history of variceal bleeding. Transfusion requirements were similar in both groups. Bleeding stopped for 12 consecutive hr during 30 hr of the study period in 31 (65%) of the somatostatin group vs. 30 (83%) of the placebo group (p = 0.06). The median time to cessation of bleeding was 2 hr in the placebo group and 3 hr in the somatostatin group. Deaths following the study period were nine (25%) in the placebo group and 15 (31%) in the somatostatin group. Within the limitations of the present study, we conclude that somatostatin was ineffective in the management of active bleeding of esophageal varices. PMID- 2573575 TI - Peripheral blood mononuclear-stem cell (PBMSC) collection in two continuous flow cell separators: yields and crosscellular contamination. PMID- 2573574 TI - A lack of genetic linkage of renin gene restriction fragment length polymorphisms with human hypertension. AB - Because renin is an important enzyme in blood pressure regulation, we studied the possibility that an alteration in the structure of the human renin gene is genetically linked to human essential hypertension or associated with levels of plasma renin activity or blood pressure. By using specific DNA probes, we have identified four polymorphisms in the human renin gene with the restriction enzymes Taq I, HindIII, Bgl I, and Bgl II. The gene location of all of these polymorphisms except for the Bgl II polymorphism has been determined, and their frequencies were initially estimated in a population of 50 random subjects. To test the clinical significance of these polymorphisms, we studied 68 persons from a large Utah pedigree with a high incidence of hypertension. Among nine relatives with hypertension, genetic linkage without recombination was ruled out by observing several obligate recombinants. We also found no significant association of the restriction fragment length polymorphisms with quantitative measurements of sitting or standing, systolic or diastolic blood pressures, or plasma renin activity in 59 untreated members of this pedigree. Although we found no genetic linkage in this set of study subjects, the characterization of the restriction fragment length polymorphisms for the renin gene may be useful in future studies of other selected pedigrees for the presence of one or more of these to be a genetic marker in hypertension. PMID- 2573576 TI - Plasma neuropeptide pattern in acute idiopathic urticaria. AB - Immediate hypersensitivity responses, as acute urticaria, produce a release of neuropeptides by nerve endings, which present specificity of recognition by mast cells, basophils and other target cells. We have measured vasoactive intestinal peptide (VIP), somatostatin, bombesin, neurotensin and beta-endorphin by radioimmunoassay in plasma extracts of 20 patients with acute idiopathic urticaria and of 20 healthy subjects. VIP- and beta-endorphin-like immunoreactivities were found to be significantly decreased with respect to controls (p less than 0.001 and p less than 0.01, respectively). On the contrary, somatostatin- and bombesin-like immunoreactivities were significantly increased (p less than 0.001 and p less than 0.05, respectively). These findings could be a reflection in blood of a raised release of somatostatin and bombesin by nerve endings in the urticaria process. Moreover, the decreased plasma levels of VIP- and beta-endorphin-like immunoreactivities could be explained by a raised specific metabolism of these peptides in the urticaria process. PMID- 2573577 TI - Terry Fox Cancer Research Workshop on Prevention Trials. PMID- 2573579 TI - Amplification of c-myc and c-erbB-2 proto-oncogenes in human solid tumors: frequency and clinical significance. AB - Molecular probes for cellular proto-oncogenes have recently been extensively used in order to search for functional and structural alterations in tumor tissues. Variable, and sometimes contradictory, results have been obtained regarding the frequency and clinical significance of amplification of the c-myc and c-erbB-2 proto-oncogenes in different series of human solid tumors. We addressed this question by performing Southern blotting analysis on 131 primary adult solid tumors of various tissues and 5 metastases of unknown origin, using molecular probes for both genes. Amplification of c-myc was found in 5 of the primary tumors, and amplification of c-erbB-2 in 5 others. In 2 tumors of the latter group, the c-erbB-2 gene was also rearranged. The distribution of these 10 tumors with regard to clinical stage and course of the disease did not point to an association between the amplification events and specific stage or prognosis. We concluded that, in this series, the amplification of both proto-oncogenes was occasional and was not a prognostic marker. PMID- 2573578 TI - Lymphocyte homing receptors and adhesion molecules in intravascular malignant lymphomatosis. AB - Intravascular malignant lymphomatosis (IML) is a highly malignant, recently recognized form of lymphoma. It is characterized by multifocal proliferation of malignant lymphocytes within small blood vessels, primarily in the central nervous system and skin, frequently resulting in circulatory disturbances. The cause of the impaired capability of the malignant lymphocytes to extravasate has remained unclear. We analyzed the presence of immunoreactivity for certain homing receptor and adhesion molecules associated with lymphocyte extravasation in 3 patients with this disease. Compared with non-neoplastic leukocytes, large malignant lymphocytes appeared either negative or only weakly positive for the leukocyte surface glycoprotein, CD18 that is the beta chain of the CDIIa/CD18 complex (lymphocyte-function associated antigen-I, LFA-I), which mediates cell-to cell adhesion of lymphocytes. On the other hand, antibody to one of the proposed ligands for this complex, intercellular adhesion molecule-I, gave positive reactivity both on lymphocytes and on endothelial cells. Further, the malignant lymphoid cells stained positively with Hermes-3 antibody, which recognizes a common structure of CD44 class of molecules involved in lymphocyte homing. It was also shown that HECA-452 antigen, a marker of high endothelial venules (HEV) supporting lymphocyte extravasation, can be synthesized by an IML patient even at the site of inflammation but it is not prerequisite for extravasation of inflammatory lymphocytes. Our results suggest that the deficiency or absence of the adhesion molecule CDIIa/CD18 may contribute to the inability of the malignant lymphoid cells to extravasate in IML, and perhaps also to the high malignancy of this form of lymphoma. PMID- 2573580 TI - Preservation of chromosome and DNA characteristics of human colorectal adenocarcinomas after passage in nude mice. AB - Twenty-seven human colorectal adenocarcinomas were implanted s.c. into nude mice. A comparative study of chromosomes and DNA between fresh tumor (FT) and xenografted tumor cells (XT) could be performed in 9 cases. Losses or deletions were consistently found in FTs as well as in XTs, particularly on chromosomes 17 and/or 18. This was correlated with loss of heterozygosity for these chromosomes. Comparison between corresponding FT and XT karyotypes showed great similarities. However, the tendency toward polyploidization, which exists in FTs, appeared to be more pronounced in XTs. Passage in nude mice makes it possible to repeat cytogenetic analyses in order to obtained interpretable metaphases. Xenografting not only increases the number of tumorigenic cells, but also eliminates human normal stromal or blood cells and gives unambiguous data on allelic loss. PMID- 2573581 TI - Treatment with P-chloroamphetamine enhances the development of neuroleptic induced orofacial dyskinesias in the rat. PMID- 2573582 TI - Increased incidence and severity of neuroleptic-induced movement disorder in pinealectomized rats. AB - The effects of prior pinealectomy on neuroleptic-induced perioral movements were examined in male Sprague-Dawley rats. Treatment with haloperidol resulted in significantly more severe movement disorder in pinealectomized rats than in unoperated control rats. Subsequent administration of melatonin (4 mg, i.p.) was associated with a nonsignificant reduction of the severity of movements in the pinealectomized rats within one hour. We conclude that pinealectomy facilitates the onset of neuroleptic-induced perioral movements, and suggest that impaired melatonin secretion may be implicated in the pathophysiology of tardive dyskinesia in humans. PMID- 2573583 TI - The ways of endocytosis. PMID- 2573584 TI - Modulation of transglutaminase by retinoic acid in liver cells. PMID- 2573585 TI - D-amino-acid oxidase from yeast. PMID- 2573586 TI - Some disease-associated ancestral haplotypes carry a polymorphism of TNF. AB - We describe here an Nco I restriction fragment length polymorphism of tumor necrosis factor carried by the 8.1 (HLA-A1,B8,BfS,C4AQ0,C4B1,DR3) and the 44.1 (HLA-B44,BfS,C4A3,C4BQ0,DR4) ancestral haplotypes associated with complications of rheumatoid arthritis. By examining multiple examples of these and other ancestral haplotypes it was seen that 8.1 and 44.1 ancestral haplotypes yield fragments of approximately 5.5 kb while many other ancestral haplotypes carry fragments of approximately 10.5 kb. The polymorphism is associated with the ancestral haplotype rather than the HLA-B or -DR allele defined by conventional serology. PMID- 2573587 TI - Comparative mapping of the human major histocompatibility complex in different racial groups by pulsed field gel electrophoresis. AB - The molecular map of the human major histocompatibility complex was examined in multiple examples of various Caucasoid and Japanese major histocompatibility complex supratypes using pulsed field gel electrophoresis. Extensive differences in restriction fragment lengths were observed. However, each supratype showed specific genomic characteristics including deletions, duplications, or insertions supporting the hypothesis that these supratypes are markers of conserved ancestral haplotypes. Some of the gene arrangements are consistent with the deletions or duplications previously described or suggested by conventional DNA techniques and protein typing, while others have not been recognized previously. Characterization of the gene organization within disease-associated ancestral haplotypes will provide new insights into the functional role and evolution of the major histocompatibility complex. PMID- 2573588 TI - Restriction fragment length polymorphism of HLA-DRw53 detected in South African blacks and individuals of mixed ancestry. AB - The HLA-DRw53 specificity has not until now been shown to demonstrate polymorphism. We have studied 33 DRw53 haplotypes, comprising 19 DR4, 10 DR7, and 4 DR9 haplotypes, from 6 homozygous typing cells, 11 families, and 8 random individuals. All the subjects studied were South African blacks or of mixed ancestry (Cape Coloureds), with the exception of four homozygous typing cells from whites. The DNA was digested with TaqI and, after Southern blotting, was hybridized with a full-length DRB cDNA probe. Fragments correlating with DR4 (5.5 kb), DR7 (4.0 kb), and DR9 (4.1 kb) were observed. Two fragments of 14.5 and 2.8 kb correlated with DRw53. In addition, two pairs of fragments demonstrated a diallelic pattern, which is likely to correlate with a polymorphism of the DRB4 (DRw53) gene, since one or other of the two patterns was observed in all cells carrying the DRw53 specificity. The first allelic pattern, called DRw53a, was characterized by the presence of 7.5- and 2.6-kb fragments, while the second pattern, called DRw53b, had 5.8- and 2.7-kb fragments. DRw53a occurred in 10 of the 19 DR4 haplotypes and 7 of the 10 DR7 haplotypes. All three DR9,DQw2 haplotypes were also associated with DRw53a. These findings may have important implications for disease associations and the use of unrelated donors for organ transplantation. PMID- 2573589 TI - Allelic T-cell receptor alpha complexes have little or no influence on susceptibility to type 1 diabetes. AB - We performed a multiple-affected-sib study to determine if T-cell receptor alpha chain alleles affect susceptibility to insulin-dependent diabetes mellitus. Restriction fragment length polymorphisms were used to follow the segregation of allelic T-cell receptor alpha complexes within the families. The segregation of T cell receptor alpha alleles in 29 multiplex families revealed no significant tendency for affected sibs to share T-cell receptor alpha-chain alleles more often than would be expected by chance alone (p greater than 0.2). In contrast, the same type of analysis for HLA alleles easily detected the well-known linkage of insulin-dependent diabetes mellitus susceptibility to the HLA complex (p = 0.003). We suggest that the importance of HLA alleles in insulin-dependent diabetes mellitus susceptibility and the lack of importance of T-cell receptor alpha alleles result from the different strategies by which HLA and T-cell receptor molecules achieve antigen-binding diversity: multiple loci and allelic diversity in the case of HLA; combinatorial, junctional, and N-region diversity in the case of the T-cell receptor. In this paper we also describe three new restriction fragment length polymorphisms of the T-cell receptor alpha complex and a new method for testing the significance of linkage in multiple-affected-sib studies. PMID- 2573590 TI - Restriction fragment length polymorphism in HLA class II genes of Latin-American Caucasian celiac disease patients. AB - In the present study Latin-American celiac disease patients were analyzed for the frequency of certain HLA class II restriction fragment length polymorphisms in order to investigate whether they exhibited the normal associated alleles or showed unusual class II variants. A DPB/RsaI 4.0-kb fragment that was shown to be significantly increased among North Americans celiac disease patients of the DR3,DQw2 haplotype was found with similar frequency in Latin-American control and celiac disease individuals. A DPA/BglII 3.7-kb fragment previously shown to be increased among British celiac disease patients was also present with similar frequency among Latin-American control and celiac disease individuals. These results show that the frequency of the HLA-DP region-derived restriction fragment length polymorphisms linked to celiac disease differs between Caucasian populations of different ethnic backgrounds (Anglo-Saxon and Latin-American). On the other hand, DNA samples from 13 patients and 14 controls bearing the DR5/DR7 phenotype (which is significantly associated with celiac disease in Latin populations) were investigated for the presence of particular restriction fragment length polymorphisms disproportionally present in celiac disease individuals. No significant differences were found between controls and patients when the DNA was analyzed with 10 different restriction enzymes and probes for DRB, DQA, DQB, and DPB HLA class II sequences. PMID- 2573592 TI - Molecular genetics of C4B deficiency in IgA nephropathy. AB - The fourth component of complement (C4) occurs in two functionally distinct isotypes, C4A and C4B. The two closely linked genes are located on chromosome 6p, between HLA-B and -DR. Several reports have established complete C4B deficiency as the major genetic risk factor for IgA nephropathy (RR = 6.5; p = 0.0004). It is not clear whether this association derives from immune dysfunction related to the absent isotype or from another disease susceptibility gene closely linked to C4B. To help distinguish between these mechanisms, we examined the molecular basis of complete C4B deficiency in five patients with IgA nephropathy and eight healthy individuals. C4 and Bf protein typing were performed by immunofixation electrophoresis of plasma. Genomic DNA was digested with several restriction enzymes, chosen to produce informative restriction fragment length polymorphisms (RFLPs). After electrophoresis and Southern blotting, digests were hybridized to a series of cDNA probes specific to the 5' and 3' ends of the C4 genes, the C4d region, and the adjacent 21-hydroxylase genes. Availability of DNA from family members allowed assignment of RFLPs to specific haplotypes. The 10 C4B-deficient IgA nephropathy-associated haplotypes displayed seven different protein phenotype/RFLP patterns. Three haplotypes consisted of the common C4B/21 hydroxylase deletion on the Bf*S, C4A*3, C4B*Q0 complotype. Two haplotypes were characterized by the C4A*3,2 duplication, with two C4 genes present but a C4A protein being produced by the gene at the usual C4B locus. All of the remaining haplotypes had unique Bf, C4, and 21-hydroxylase patterns. C4B-deficient IgA nephropathy patients display a variety of molecular genetic bases for their protein deficiency. This observation speaks against linkage of C4B deficiency with a locus encoding disease susceptibility and supports a primary role for the complement abnormality in this disease. PMID- 2573591 TI - DQA1 restriction fragment length polymorphisms and insulin-dependent diabetes mellitus: a BglII fragment labels a subset of B8,DR3 haplotypes uniquely associated with insulin-dependent diabetes mellitus. AB - Class II restriction fragment length polymorphism studies of 38 pedigrees with multiple cases of insulin-dependent diabetes mellitus revealed the existence of a DQA1-related polymorphism that distinguishes two kinds of HLA-B8,DR3 haplotypes. One of these, characterized by the presence of DQA1-BglII 7.20 kb, was present in all 14 examples inherited by patients and in 6 of the 12 B8,DR3 haplotypes not so inherited. The apparently complete association between the presence of this fragment and the "affected" status of B8,DR3 haplotypes (p = 0.004) was confirmed in a separate group of 26 simplex pedigrees selected for the presence of this haplotype in the respective probands (combined p less than 0.0001). PMID- 2573594 TI - Pain control in the terminally ill child at home. AB - The seriously ill or terminally ill child with cancer has received inadequate pain control in the past, partly due to physicians' and nurses' fears and misconceptions regarding the administration of effective pain medications to a child. Advances in assessment techniques in the infant and young child, as well as increasing use of pain assessment questionnaires and VAS in the older child, have mandated changes in administration of analgesia to children. It is the responsibility of the health-care team of provide adequate pain control to the ill child, using knowledgeable assessment and monitoring skills. The goal of therapy for the dying child is to maintain comfort and support the child and the family. Providing analgesia in the hospital or the home has proven safe and effective when administered either orally or parenterally, and comfort of the child is achieved. PMID- 2573593 TI - Characteristics of a human liver allograft--derived T-cell line that exhibits suppressor activity. AB - In an attempt to identify predominant cell populations that may mediate liver allograft dysfunction, the phenotypic and functional characteristics of lymphoid cells propagated from needle biopsy specimens of rejecting liver transplants were examined. In one case, a T-cell line of host phenotype propagated from a liver allograft biopsy demonstrated significant in vitro suppressor activity. This T cell line (designated JB) was maintained for almost one year in culture with medium containing human recombinant interleukin 2 and with weekly stimulation by an Epstein-Barr virus-transformed B-cell line derived from the liver donor. Repeated analyses demonstrated that the JB line was phenotypically stable and predominantly CD3+ (86-93%), CD4+ (88-96%), DR+ (96%), Leu8-, CD45R-, CD16-, with a minor CD8+ cell population (less than 5%). The JB line demonstrated proliferative responsiveness upon coculture with cells expressing disparate donor HLA antigens but no in vitro cytotoxic activity. However, JB cells significantly (greater than 90%) suppressed mixed lymphocyte reaction or phytohemagglutinin stimulation of nonautologous peripheral blood lymphocytes. Supernatants of JB cells that had been cultured alone or with irradiated (6000 rads) Epstein-Barr virus-transformed donor B cells mimicked the suppressive activity of the JB cell line, either upon addition in vitro or by transient (4 hr) pretreatment of responder cells at 20 degrees C. JB cell supernatants were nontoxic and free of tumor necrosis factor activity, and their suppressive activity was dose dependent, nondialyzable (greater than 100 kDa), not overcome by exogenous interleukin 1 or interleukin 2, and heat-resistant up to 56 degrees C. However, the suppressive activity of JB supernatants could be diminished or abrogated by treatment with high temperature (80-100 degrees C), reducing agents, trypsin, or absorption by peripheral blood lymphocytes at room temperature. The suppressive activity of JB cells and supernatants was not alloantigen-specific or major histocompatibility complex-restricted, did not shift mixed lymphocyte reaction kinetics, and was capable of inhibiting in vitro stimulation of peripheral blood lymphocytes in mixed lymphocyte reaction only when presented early in the culture. These findings provide the first evidence for a primary human allograft derived T-cell line with suppressor-effective function. PMID- 2573595 TI - Allied health perceptions of effective clinical instruction. AB - Clinical instruction is a critical component of allied health education. The purposes of this study were to identify those behaviors of clinical instructors perceived as both most effective and most hindering in facilitating learning, to identify and compare the behaviors of clinical instructors as perceived by two different allied health groups, and to categorize into meaningful domains the behaviors identified. A published 58-item questionnaire was completed by 311 clinical students and instructors from eight physical therapy and ten physician assistant programs. Results were analyzed by multivariate analysis of variance. Instructor behaviors rated as most helpful in learning included answering questions clearly, taking time for discussion and questions, and providing opportunities for practicing skills. Behaviors most hindering to learning were asking questions in an intimidating manner and correcting student errors in front of patients. Ratings were significantly different (P less than or equal to .001) between the physical therapy and physician assistant groups on 13 items, and posed important considerations for allied health educators. PMID- 2573597 TI - The N-terminal cysteine of human asparagine synthetase is essential for glutamine dependent activity. AB - Site-specific mutagenesis was used to replace the N-terminal cysteine in human asparagine synthetase by an alanine. The mutant enzyme was expressed in the yeast Saccharomyces cerevisiae, and the asparagine synthetase activity was analyzed in vitro. The mutation resulted in the loss of the glutamine-dependent asparagine synthetase activity, while the ammonia-dependent activity remained unaffected. These results confirm the existence of a glutamine amidotransfer domain with an N terminal cysteine essential for the glutamine-dependent asparagine synthetase activity. PMID- 2573598 TI - Identification of a clathrin binding subunit in the HA2 adaptor protein complex. AB - The HA2 adaptor complex, comprising alpha-, beta-, 50-kDa, and 16-kDa subunits, was partially dissociated into its constituents with 3 M urea, and the beta subunit was purified from the mixture by hydroxylapatite and affinity chromatography. The renatured beta-subunit behaves hydrodynamically as a single polypeptide of Mr approximately 128,000. In a sedimentation assay the purified beta-polypeptide co-sediments with pre-formed clathrin cages. The beta polypeptide, however, will not induce assembly of clathrin triskelia. Our results support the conjecture that the beta-type subunits (beta and beta') of the HA2 and HA2 adaptor complexes serve to attach the HA-2 adaptor complex to clathrin (Ahle, S., Mann, A., Eichelsbacher, U., and Ungewickell, E. (1988) EMBO J. 7, 919 929), while the other subunits may determine the specificity of binding to docking proteins and receptors on cytoplasmic membrane surfaces. PMID- 2573596 TI - Cardioprotective and antiarrhythmic effects of beta-blockers, propranolol, bisoprolol, and nipradilol in a canine model of regional ischemia. AB - Cardioprotective and antiarrhythmic effects of three beta-blockers with different pharmacological properties were investigated in 33 anesthetized dogs with a 2-h coronary occlusion. Dogs were divided into 4 groups and received physiological saline or one of the following drugs using a 10-min infusion at 25 min before the occlusion: saline or control (n = 12), propranolol (0.3 mg/kg, n = 7), bisoprolol (0.05 mg/kg, n = 7), and nipradilol (0.2 mg/kg, n = 7) groups. Blood pressure did not significantly differ among the 4 experimental groups throughout the entire observation period. On the contrary, the postocclusion change (fall) in heart rate from the preocclusion value was significantly (P less than 0.05-0.01) greater in the drug-treated groups than in the control group. Each of the beta blockers effectively prevented the development of ventricular arrhythmias associated with the 2-h coronary occlusion. In terms of assessing a cardioprotective effect, the respiratory control index and rate of oxygen consumption in State III in mitochondria, and lysosomal enzyme activities (N acetyl-beta-glucosaminidase or beta-glucuronidase) in myocardial tissues, all prepared from both ischemic and non-ischemic areas, were measured using the respective, established methods. The 2-h coronary occlusion induced a mitochondrial dysfunction and leakage of lysosomal enzymes in the control group, whereas each beta-blocker significantly (P less than 0.05-0.01) protected mitochondria against ischemia and prevented the lysosomal enzyme leakage. The results indicate that the antiarrhythmic effects of beta-blockers on ischemic myocardium are, at least in part, due to their cardioprotective action, and these effects appear to be unrelated to the ancillary pharmacological properties of these drugs. PMID- 2573599 TI - Site-specific mutants of oncomodulin. 1H NMR and optical stopped-flow studies of the effect on the metal binding properties of an Asp59----Glu59 substitution in the calcium-specific site. AB - High resolution 1H nuclear magnetic resonance spectroscopy and optical stopped flow techniques have been used to study the metal binding properties of a site specific mutant of bacterial recombinant oncomodulin in which glutamate has replaced a liganding aspartate at position 59 in the CD calcium-binding site. In particular we have followed the replacement of calcium by lutetium in bacterial recombinant oncomodulin and D59E oncomodulin to provide a measure of the protein's preferences for metal ions of different ionic radii. The result of the Asp----Glu substitution is to make the mutant oncomodulin more similar to rat parvalbumin in terms of its relative CD- and EF-domain affinities for lutetium(III), that is to increase its affinity for metal ions with smaller ionic radii. This finding supports the original hypothesis that the presence of Asp at sequence position 59 is an important factor in the reduced preference of the CD site of oncomodulin for smaller metals such as magnesium (Williams, T. C., Corson, D. C., Sykes, B. D., and MacManus, J. P. (1987) J. Biol. Chem. 262, 6248 6256). However, our studies show that both the CD and the EF sites are affected by this single residue substitution suggesting that many factors play a role in the metal binding affinity and interaction between the two sites. PMID- 2573600 TI - Transglutaminase-mediated cross-linking of fibrinogen by human umbilical vein endothelial cells. AB - The interaction of endothelial cells with soluble or substrate-immobilized 125I labeled fibrinogen (125I-FGN) was analyzed. Binding experiments involved incubation of 125I-FGN with cell suspensions at 4 degrees C. Bound ligand was quantitated by centrifugation of cells through silicone oil followed by scintillation analysis of the cell pellet. Calcium-dependent binding of 125I-FGN reached a maximum after 3 h and represented about 60% of the total. Half-maximal saturation occurred at 60 nM, and about 9 x 10(4) molecules were bound/cell at saturation (approximately 100 nM). Calcium-dependent binding was completely inhibited by unlabeled fibrinogen, partially inhibited by a monoclonal antibody (7E3) against glycoprotein IIb-IIIa, but not inhibited by fibrinogen fragments D or E, an anti-glycoprotein IIIa polyclonal antibody, or the Arg-Gly-Asp-Ser tetrapeptide. In contrast, the Arg-Gly-Asp-Ser tetrapeptide as well as the monoclonal antibody 7E3 markedly inhibited attachment of endothelial cells to substrate-immobilized fibrinogen, whereas fragment D or E did not. Both in suspension and monolayer, the 125I-FGN underwent cross-linking involving principally the A alpha chain. The transglutaminase inhibitors putrescine, histamine, and cystamine interfered with 125I-FGN binding and cross-linking by suspended cells. Since cross-linking in suspension was limited to bound 125I-FGN and since transglutaminase activity was not detectable in the binding buffer, cross-linking may have been mediated by a cell-associated transglutaminase. PMID- 2573601 TI - Functional water channels are present in clathrin-coated vesicles from bovine kidney but not from brain. AB - Targeting of water channels in renal epithelia may involve trafficking of clathrin-coated vesicles. We have isolated and measured the osmotic water permeability (Pf) of purified clathrin-coated vesicles from bovine kidney cortex and inner medulla, and bovine brain, a tissue not expected to contain "water channels." Brain-coated vesicles had a diameter of 80 nm in negatively stained preparations. Pf was measured by a stopped-flow light scattering technique. In brain-coated vesicles, water transport was functionally homogeneous with a low Pf of 0.0016 +/- 0.0001 cm/s (seven preparations, 23 degrees C). Pf was independent of osmotic gradient size (25-300 mOsm), not inhibited by mercurials, and not altered by removal of the clathrin coat. The activation energy (Ea) for Pf was high (11 +/- 1 kcal/mol less than 34 degrees C, 17 +/- 2 kcal/mol greater than 34 degrees C). Therefore, water channels are absent from brain-coated vesicles. In contrast, there were two functional populations of vesicles in coated vesicle preparations from both kidney cortex and medulla. One population of vesicles had low water permeability and no water channels, whereas a second population had high Pf (0.02 cm/s, 21 degrees C) that was inhibited by HgCl2, and low Ea (2-3 kcal/mol). The fraction of vesicles with high Pf was 52 +/- 3% (S.D., n = 3, cortical vesicles) and 26 +/- 3% (medullary vesicles). These results provide evidence that functional water channels are not present in clathrin-coated vesicles from the brain, whereas they are found in a population of coated vesicles from kidney cortex and medulla, tissues in which water channels are recycled between the plasma membrane, and an intracellular compartment. PMID- 2573602 TI - Atrial natriuretic peptide-dependent phosphorylation of smooth muscle cell particulate fraction proteins is mediated by cGMP-dependent protein kinase. AB - The atrial natriuretic peptide (ANP) stimulates cGMP production and protein phosphorylation in a particulate fraction of cultured rat aortic smooth muscle cells. Three proteins of 225, 132, and 11 kDa were specifically phosphorylated in response to ANP treatment, addition of cGMP (5 nM), or addition of purified cGMP dependent protein kinase. The cAMP-dependent protein kinase inhibitor had no effect on the cGMP-stimulated phosphorylation of the three proteins but inhibited cAMP-dependent phosphorylation of a 17-kDa protein. These results demonstrate that the particulate cGMP-dependent protein kinase mediates the phosphorylation of the 225-, 132-, and 11-kDa proteins. The 11-kDa protein is phospholamban based on the characteristic shift in apparent Mr from 11,000 to 27,000 on heating at 37 degrees C rather than boiling prior to electrophoresis. ANP (1 microM) increased the cGMP concentration approximately 4-fold in the particulate fractions, from 4.3 to 17.7 nM, as well as the phosphorylation of the 225-, 132-, and 11-kDa proteins. In contrast, the biologically inactive form of ANP, carboxymethylated ANP (1 microM), did not stimulate phosphorylation of any proteins nor did the unrelated peptide hormone, angiotensin II (1 microM). These results demonstrate the presence of the cGMP-mediated ANP signal transduction pathway in a particulate fraction of smooth muscle cells and the specific phosphorylation of three proteins including phospholamban, which may be involved in ANP-dependent relaxation of smooth muscle. PMID- 2573603 TI - Identification, characterization, and purification of two mammalian stress proteins present in mitochondria, grp 75, a member of the hsp 70 family and hsp 58, a homolog of the bacterial groEL protein. AB - We describe the identification, characterization, and purification of two members of the mammalian stress protein family, both of which are shown to be components of the mitochondria. The first, with an apparent mass of 58,000 daltons, is a constitutive protein whose synthesis increases in cells exposed to elevated temperatures and/or amino acid analogs and is therefore referred to as heat shock protein hsp 58 (hsp 58). The second, with an apparent mass of 75,000 daltons, is also a constitutive protein whose synthesis is increased in cells following glucose deprivation or exposure to either a calcium ionophore or 2-deoxyglucose and therefore represents a member of the so-called glucose-regulated proteins (grp 75). In cells treated with the potassium ionophore, nonactin, both hsp 58 and grp 75 were observed to accumulate in precursor form. Since nonactin has been reported to specifically inhibit the processing of cytoplasmic precursor proteins destined for the mitochondria, we investigated whether mature hsp 58 and grp 75 were components of the mitochondria. Mitochondria were isolated from rat liver and shown to contain both hsp 58 and grp 75. Indirect immunofluorescence using antibodies specific to either hsp 58 or grp 75 confirmed their presence within mitochondria. Proteolytic digestion experiments with intact mitochondria indicated that both proteins were not accessible to external proteolytic attack, suggesting that they are not exposed on the cytoplasmic face of the outer membrane. Based on a variety of biochemical and immunological criteria, grp 75 is shown to be a member of the hsp 70 family of stress proteins, while hsp 58 represents the mammalian equivalent of the bacterial groEL protein. Procedures for the purification of both hsp 58 and grp 75 are presented. The possible biochemical role of these two mitochondrial stress proteins is discussed in relation to the known biochemical function of their related stress protein counterparts. PMID- 2573604 TI - O-linked glycosylation of rat renal gamma-glutamyltranspeptidase adjacent to its membrane anchor domain. AB - Large domains rich in serine and threonine, that are likely to exhibit clusters of O-linked oligosaccharides, have been reported adjacent to the anchor of several cell surface proteins. No such domain is evident in the primary sequence of rat renal gamma-glutamyltranspeptidase. However, papain treatment of the amphipathic enzyme (Triton-purified gamma-glutamyltranspeptidase, T gamma GT), pretreated with galactose oxidase and NaB3H4 (Frielle, T., and Curthoys, N. P. (1983) Biochemistry 22, 5709-5714), yields the hydrophilic enzyme (papain-treated Triton-purified gamma-glutamyltranspeptidase, PT gamma GT) and a labeled peptide which contains both the amino-terminal membrane anchor and the sequence Pro27 Thr28-Thr29-Ser30. Since [3H]galactose was identified in this peptide, the presence of O-linked oligosaccharides was investigated. Carbohydrate analysis is consistent with the presence of two simple O-linked oligosaccharides on T gamma GT and one on PT gamma GT. Lectin blot analysis of T gamma GT and PT gamma GT was carried out after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The small subunits of both T gamma GT and PT gamma GT and the large amphipathic subunit of T gamma GT all react with the peanut agglutinin lectin, but the large subunit of PT gamma GT exhibits no such reactivity. The reactivity with PNA is consistent with the presence of one oligosaccharide with the structure galactose beta 1-3N-acetylgalactosamine alpha 1-Ser/Thr attached to each subunit of T gamma GT. The papain-sensitivity of the oligosaccharide from the larger subunit is consistent with O-glycosylation at the Thr28-Thr29-Ser30 sequence. The results of lectin blot analysis with wheat germ agglutinin imply that the content of N linked oligosaccharides is unaffected by papain treatment of the transpeptidase. These data represent the first direct evidence for O-glycosylation of a microvillar hydrolase at a site immediately adjacent to the membrane anchor and indicates that even small clusters of Thr and Ser can be O-glycosylated. Isolated O-linked oligosaccharides may have functional significance since single Ser and Thr residues are consistently found near the membrane anchor of many cell surface proteins. PMID- 2573606 TI - Stereospecific liquid chromatographic analysis of racemic adrenergic drugs utilizing precolumn derivatization with (-)-menthyl chloroformate. PMID- 2573605 TI - Characterization of glutamate dehydrogenase isoproteins purified from the cerebellum of normal subjects and patients with degenerative neurological disorders, and from human neoplastic cell lines. AB - Glutamate dehydrogenase (GDH) was purified to homogeneity from cerebellar tissue of three normal subjects and seven patients with four distinct types of degenerative neurological disorders. Nonequilibrium pH gradient gel electrophoresis showed that the purified enzyme consists of four major isoproteins designated GDH 1, 2, 3, and 4. With one exception, the relative abundance and isoelectric points of the GDH isoproteins decrease and the molecular weights increase progressively going from isoprotein 1 to isoprotein 4. The enzyme isolated from the brain of one patient with a variant form of multiple system atrophy displayed marked reduction of GDH isoprotein 1. The Km values of the patients' GDH for alpha-ketoglutarate, glutamate, NADH, and NADPH were significantly increased as compared to GDH obtained from normal and neurologic control subjects. In addition, glutamate levels were reduced markedly in the patient's cerebellum. Pulse-chase studies have shown that both the human hepatoma HepG2 and the human glioma U373 cell lines synthesize exclusively GDH isoprotein 2. The different GDH isoproteins do not have a precursor-product relationship and may represent products of different GDH mRNA species. PMID- 2573607 TI - Rapid and sensitive column liquid chromatographic determination of sotalol in plasma. PMID- 2573608 TI - Determination of the partial benzodiazepine receptor agonist Ro 16-6028 in plasma by capillary gas chromatography with nitrogen-selective detection after conversion into the ethyl ester derivative. AB - A highly sensitive capillary gas chromatographic method was developed to determine plasma levels of a novel partial benzodiazepine receptor agonist in man following the very low therapeutic doses required for anxiolysis. The compound was isolated from plasma by liquid-liquid extraction at basic pH, converted into the ethyl ester analogue by a two-step procedure, separated from plasma constituents by capillary gas chromatography and quantified by means of nitrogen selective detection. Because of the thermolabile tert.-butyl ester function, the agonist could not be gas chromatographed without degradation. Formation of the far more stable ethyl ester analogue was achieved by treatment with hydrogen chloride in ethanol, followed by an ethylation step with diazoethane. The high sensitivity of the new method (about 100 pg/ml, using 1-ml plasma specimens) allowed the monitoring of plasma levels of the agonist for up to 8 h (about three elimination half-lives) after a single 0.1-mg oral dose to human volunteers. The practicability of the procedure was demonstrated by the analysis of more than 600 plasma samples from clinical studies performed with human volunteers. PMID- 2573609 TI - Capillary gas chromatography of trihexyphenidyl, procyclidine and cycrimine in biological fluids. AB - A sensitive (50-100 pg/ml) method is described for the analysis of the anticholinergic drugs cycrimine, procyclidine and trihexyphenidyl by capillary gas chromatography with flame thermionic detection. Since these anticholinergic drugs are frequently administered in combination with antipsychotic medication for the treatment of mental illness, the potential interference by antipsychotic drugs in this assay was examined. No interference was observed from a series of antipsychotic drugs in the quantitation of cycrimine, procyclidine or trihexyphenidyl. The use of this technique to study trihexyphenidyl pharmacokinetics in man is described. PMID- 2573610 TI - Determination of an oxadiazole-substituted 1,4-benzodiazepine in plasma by high performance liquid chromatography with ultraviolet detection and by a radioreceptor assay. AB - A high-performance liquid chromatographic (HPLC) method for the determination of an oxadiazole-substituted 1,4-benzodiazepine [3-(5-cyclopropyl-1,2,4-oxadiazol-3 yl)-5,6-dihydro-5-methyl-6-oxo -4H-imidazo[1,5a] [1,4]benzodiazepine] in plasma has been developed and compared with a radioreceptor assay. The results given by the two methods were in good agreement, with detection limits of ca. 1 ng/ml (signal-to-noise ratio = 3). The radioreceptor method is preferred for the monitoring of toxicological and other well controlled studies, while HPLC is preferred where greater specificity is essential. Further, the HPLC assay is applicable over a much wider concentration range. PMID- 2573611 TI - High-performance liquid chromatographic method for a clozapine analogue, CGS 13429, and its N-oxide and desmethyl metabolites. PMID- 2573612 TI - Reliable isolation of human immunodeficiency virus from cultures of naturally infected CD4+ T cells. AB - A procedure for reliable and reproducible isolation of human immunodeficiency virus (HIV) from cultures of CD4+ T cells from healthy HIV seropositive individuals is described. Using immunomagnetic cell separation techniques, CD4+ T cells were positively selected from blood or peripheral blood mononuclear cells from 34 HIV infected individuals in CDC group II. The cells were stimulated with beads coated with a monoclonal antibody specific for the T cell receptor alpha/beta dimer and cultured in medium containing recombinant IL 2. HIV was isolated from 100% of the 41 cultures from 34 individuals. As this culture system allows reproducible isolation of HIV from cultures of naturally infected CD4+ T cells in the absence of other autologous or allogeneic cells, it may provide a good test system for the study of factors affecting the replication of HIV at low multiplicities of infection. PMID- 2573613 TI - Comparison of dopamine and fenoldopam effects on renal blood flow and prostacyclin excretion in normal and essential hypertensive subjects. AB - We recently reported that a low dose dopamine (DA) infusion in normal subjects increased renal blood flow (RBF) via prostacyclin (PGI2) formation without changes in PGE2 levels. The present study explores whether this mechanism is mediated by the DA1 receptor and evaluates the effect of DA on RBF and PGs in subjects with essential hypertension (EH). A low dose of DA (1 microgram/kg.min), which previously did not alter hemodynamics in normal subjects was infused into eight patient with EH to determine the role of DA stimulation in hypertensives. To assess the effect of DA1 stimulation, fenoldopam, a selective DA1 agonist, was infused (0.1 microgram/kg.min) into 10 normal and 10 hypertensive patients. Fenoldopam, unlike DA, significantly decreased diastolic blood pressure in hypertensives (96 +/- 3 to 85 +/- 2 mm Hg; P less than 0.01) along with a significant increase in pulse rate (68 +/- 2 vs. 73 +/- 2 beats/min; P less than 0.01). RBF measured by para-aminohippurate clearance increased only in normals during fenoldopam infusion from 1185 +/- 71 to 1533 +/- 84 L/min.m2 (PGI(2)01), and this was associated with an increase in PGI2 (6-keto-PGF1) excretion (149 +/- 19 vs. 214 +/- 32 ng/g creatinine; P less than 0.02). These effects of fenoldopam were similar to DA effects on RBF and PGI2 excretion in normals. In contrast, in hypertensive subjects, neither fenoldopam (867 +/- 113 vs. 1054 +/- 177 L/min.m2; P greater than 0.1) nor DA (1098 +/- 85 vs. 1061 +/- 101 L/min.m2; P greater than 0.1) increased RBF. Similarly, neither the DA nor the fenoldopam infusion stimulated PGI2 excretion in the hypertensives. The fenoldopam infusion in the hypertensives produced a significant natriuresis (22 +/- 3 to 49 +/- 9 mmol/3 h; P less than 0.05). Similar effects on Na+ excretion in this group were noted during DA infusion (17 +/- 3 to 36 +/- 3 mmol/3 h; P less than 0.05), suggesting that DA-induced natriuresis is not directly linked to DA-induced changes in RBF or PG excretion. The present study shows that in normal subjects, fenoldopam, a specific DA1 agonist, like DA, stimulates renal PGI2 release and RBF. In contrast, neither DA nor fenoldopam alters PGI2 or RBF in patients with EH, suggesting an alteration of dopaminergic tone in some hypertensives that is characterized by a defect in DA1 receptor sensitivity. PMID- 2573614 TI - Beta-adrenergic modulation of growth hormone (GH) autofeedback on sleep associated and pharmacologically induced GH secretion. AB - To determine whether GH feedback affects both induced and spontaneous GH secretion and to explore its neurotransmitter mediation, we assessed the effects of 6-h GH infusions (0.55-5.5 micrograms/m2/min) on sleep-associated and GH releasing hormone (GHRH)-, insulin hypoglycemia-, and arginine-stimulated GH secretion and their modulation by beta-adrenergic blockade in normal men. GH infusions initiated 2 h before the expected onset of sleep produced a dose dependent inhibition of GH secretion. GH infusions (0.55 micrograms/m2/min) initiated 4 h before the stimuli inhibited the GH response to each, but did not alter the TSH response to TRH. Propranolol infusion (80 micrograms/min) started 2 h before the onset of sleep or the stimulus enhanced GH responses to GHRH and insulin alone and in the presence of GH. In contrast, propranolol neither enhanced the GH responses to arginine or sleep nor reversed the inhibitory effects of GH. The negative feedback effect of GH to both physiological and pharmacological stimuli of GH secretion indicates that it is most likely mediated by both stimulation of somatostatin and inhibition of GHRH release. The effects of beta-adrenergic blockade suggest an inhibition of somatostatin release, although the complex interaction of GH and propranolol implies that they act through dissimilar mechanisms. PMID- 2573615 TI - Expression of transcripts encoding a parathyroid hormone-related peptide in abnormal human parathyroid tissues. AB - A PTH-related peptide (PTHRP) has been identified and its cDNA cloned from human tumors associated with the syndrome of humoral hypercalcemia of malignancy. The human PTHRP gene has been recently isolated and found to be a complex transcriptional unit using multiple promoters and containing alternatively spliced 3' exons which result in three mRNA classes, each class encoding a PTHRP with a unique carboxy-terminus. The PTHRP gene appears to be expressed in a number of normal tissues, and PTHRP transcripts have been previously reported to be overexpressed in a small sample of human parathyroid adenomas. In the present study we surveyed RNA prepared from a total of 60 abnormal human parathyroid glands for PTHRP gene expression using a combination of Northern blotting and RNase protection techniques. Apparent overexpression of PTHRP mRNA was observed in two thirds of parathyroid adenomas, whereas no overexpression was found in 7 examples of sporadic primary hyperplasia, 5 examples of secondary hyperplasia, and 3 examples of parathyroid carcinoma. Apparent overexpression was also observed in 1 of 4 cases of multiple endocrine neoplasia type 1, 1 of 2 examples of multiple endocrine neoplasia type 2, and 1 gland considered to represent tertiary hyperparathyroidism. Northern analysis of poly(A)+ RNA prepared from three representative adenomas using region-specific probes indicated that two putative promoters are used and revealed a pattern of preferential splicing of transcripts to include the most distal 3' exon. These findings suggest that the PTHRP gene is commonly overexpressed in adenomatous parathyroid glands, but not in sporadic primary hyperplasia, that this overexpression does not seem to be dependent on the use of a single specific promoter, and that adenomatous parathyroid cells appear to preferentially use one of several alternative splicing pathways. It is presently not known whether PTHRP is secreted by abnormal parathyroid tissues and, if so, in what form. PMID- 2573616 TI - DNA probe for the identification of Histoplasma capsulatum. AB - A 1.85-kilobase HindIII nuclear DNA probe from Histoplasma capsulatum G217B detected polymorphic restriction fragments within whole-cell DNA from different clinical isolates of H. capsulatum, consistent with the previous system of classification. The probe failed to hybridize to DNA from Blastomyces dermatitidis, Candida spp., Saccharomyces cerevisiae, Sepedonium chrysospermum, and Chrysosporium keratinophilum under low-stringency conditions and therefore may have value as a diagnostic reagent to identify H. capsulatum. PMID- 2573617 TI - Thymic stromal elements contain an anterior pituitary hormone-stimulating activity. AB - Conditioned medium from thymic reticular monolayers displayed time-dependent accumulations of a concentration-responsive pituitary hormone-releasing activity that has been named thymic neuroendocrine-releasing factor (TNRF). Dopamine blocked and somatostatin (SRIF) attenuated TNRF-induced prolactin (PRL) release. Conversely, SRIF had no effect on TNRF-induced growth hormone (GH) release. TNRF potentiated thyrotropin-releasing hormone (TRH)-stimulated PRL release and was additive to the effects of GH-releasing hormone (GHRH) on GH release. Anterior pituitary cells perifused with TNRF responded with immediate, sustained and reversible increases in hormone release. Partial purification revealed this activity to be greater than 10,000 in molecular weight. These data suggest that the thymus may affect pituitary function. PMID- 2573618 TI - The beta-adrenergic agonist isoproterenol suppresses experimental allergic encephalomyelitis in Lewis rats. AB - Treatment with the beta-adrenergic agonist isoproterenol suppresses clinical and histological experimental allergic encephalomyelitis in Lewis rats. The effect of isoproterenol treatment is greater when the drug is given from the time of immunization through the acute phase of the illness or from 8 to 14 days post immunization than when given for the first 7 days after immunization. PMID- 2573619 TI - Emotive imagery and children's night-time fears: a multiple baseline design evaluation. AB - Emotive imagery is a variant of systematic desensitization that has been used in the treatment of children's excessive fears. The purpose of this study was to evaluate the efficacy of emotive imagery as a fear reduction procedure using a multiple baseline design across subjects. Three children (6, 8 and 11 years old) with night-time fears participated in the study. The children showed marked behavioral improvements on a test for darkness toleration that was administered on alternate days in their homes. However, they reported experiencing very little fear in these tests. Over the intervention period fewer night-time behavioral disturbances were observed for two subjects. Parents expressed some concern about the darkness test but were satisfied with emotive imagery as a fear reduction procedure. As children's phobias are characterized by multiple processes, we recommend a combination of treatment procedures in clinical practice. PMID- 2573620 TI - Treatment programs for severe night-time fears: a methodological note. AB - A multiple baseline design across subjects was used to examine the efficacy of a treatment package consisting of relaxation, reinforcement, and cognitive self instruction in the reduction of severe night-time fears in six children. Although the disruptive bedtime behaviors of five of the six children were reduced, the multiple baseline analysis revealed that changes were not solely due to treatment. For children with extended baselines, improvement preceded treatment. The implications of these findings for the treatment of fear of the dark and multicomponent treatment programs are discussed. PMID- 2573621 TI - Comparative quality assessment in immunocytochemistry: pilot study of CD15 staining in paraffin wax embedded tissue in Hodgkin's disease. AB - The feasibility of comparative quality assessment studies in immunocytochemistry was examined. The reactions of three CD15 antibodies--anti-Leu M1, DM1, and Tu9- were examined in paraffin wax sections in Hodgkin's disease under a variety of different fixation and pre-treatment conditions, using four immunochemical detection techniques. All three antibodies stained Reed-Sternberg cells, but DM1 could be used at slightly higher dilutions to achieve comparable results. Tissue fixed in formol sublimate showed the most intense staining reactions, and formol saline and neutral buffered formalin gave relatively poor results. Although neuraminidase pre-treatment improved staining, its routine use is probably contraindicated by its high cost. Trypsinisation has some value for sections of tissue fixed in formol saline and neutral buffered formalin. The avidin-biotin complex technique produced the best results, but indirect immunoperoxidase produced acceptable results, is technically easier to perform, and is less expensive. It is concluded that information regarding variations in techniques and commercially available reagents, which may be of use in routine diagnostic histopathology, can be obtained by comparative quality assessment studies of this type. PMID- 2573622 TI - Photo-onycholysis caused by clorazepate dipotassium. PMID- 2573623 TI - Transglutaminase expression in rat parotid gland after isoproterenol stimulation. AB - Transglutaminases (E.C. 2.3.2.13) are calcium-dependent enzymes that catalyze the covalent cross-linking of proteins, and occur in multiple molecular forms in a variety of tissues. Distribution of each form of transglutaminase varies with different tissues. Studies were undertaken to characterize the form of transglutaminase expressed in rat parotid gland, and to examine a possible physiological role for the enzyme. It was found that chronic treatment of rats with the beta-adrenergic agonist isoproterenol (IPR) resulted in the induction of parotid transglutaminase activity. The properties of this transglutaminase appeared to be distinct from those of the well-characterized guinea pig liver cytosol transglutaminase (TGase C). The findings that protein polymerization (observed on SDS-PAGE) and incorporation of radioactive putrescine, a polyamine, into protein occur in the presence of exogenous transglutaminase and calcium indicated that certain rat parotid salivary proteins are or could be substrates for this enzyme. Analysis of proteolytic digests of rat parotid salivary proteins on an amino acid analyzer and by high-performance liquid chromatography also indicated that these salivary proteins contain gamma-glutamyl derivatives of primary amines (e.g., polyamines or lysine), post-translational products of transglutaminase catalysis. The possible physiological function of this enzyme in the oral cavity might be stabilization of proteinaceous structures during normal oral homeostasis and/or woundhealing. PMID- 2573624 TI - Beta-adrenergic blockade and acute myocardial infarction. PMID- 2573625 TI - Thyroglobulin, thyrotropin and thyrotropin binding inhibiting immunoglobulins assayed at the withdrawal of antithyroid drug therapy as predictors of relapse of Graves' disease within one year. AB - "Sensitive" thyrotropin (TSH), thyroglobulin (TG) and even thyrotropin binding inhibiting immunoglobulins (TBII) assays are now widely available. The objective of the present study was to determine the most accurate of these three parameters to predict the relapse of Graves' disease during the year following treatment discontinuation and to evaluate whether the assay of three markers is able to improve the prediction. TSH, TG and TBII were measured in the sera of 67 Graves' disease patients after at least 12 months of medical treatment. In 52 patients, TBII had also been determined before the beginning of the medical treatment. Under treatment, all the patients were clinically and biologically euthyroid, but in 9 goitrous patients it was impossible to lower the doses of carbimazole without an immediate relapse. The TSH levels of these 9 patients were still low in all cases but one; TG and TBII levels were abnormal in all. In the other 58 patients, the treatment was discontinued; 22 relapsed within one year, more frequently when a goiter was present. The most reliable parameter for the prediction of relapse was found to be TBII, as its specificity was high (94.5%), although its sensitivity was poor (45%); TG was more sensitive (64%) but far less specific (57%); TSH and "initial" TBII appeared to be of a little interest. When TBII was elevated prior to the withdrawal of treatment, the determination of TG was useful: abnormal values of both TBII and TG were always associated with a relapse. When TBII testing was negative, the relapse risk fell to 0.26, and to 0.08 when three criteria were matched: no goiter, negative TBII, normal TG. PMID- 2573626 TI - Thyroid growth stimulating activity in highly purified IgG-fractions of patients with nonimmune thyroid diseases. AB - Two different proliferation assays have been used to measure the proliferative potential of IgG-fractions from 57 patients with nontoxic goiter of an iodine deficient area: primary human thyroid epithelial cells (TEC) and the thouroughly investigated FRTL-5 cell line. IgG-fractions from patients with nontoxic goiter (n = 30), nontoxic recurrent goiter (n = 8), toxic-nodular goiter (n = 15) and carcinoma of the thyroid (n = 4) were highly purified on DEAE-Sepharose and additionally Protein A-Sepharose in some cases. The two proliferation assays gave contradictory results: primary cultures of human thyroid epithelial cells (TEC) could not be stimulated by any of the patient's IgG-fractions nor by bTSH. The FRTL-5 cells, however, were stimulated with 10 microU/ml bTSH by 326% +/- 96% (range: 222% - 497%, p less than 0.001). In one experimental series, 72% of all patients exceeded mean + 2 SD of normal controls, when the stimulation index was referred to the effect of bTSH (NTG: 77%, Rec. G.: 88%, Tox. G.: 53%, Ca. thyroid: 75%). With a different method of calculation - stimulation index referred to the basal value - the number of patients above mean + 2 SD of normal controls decreased to 30% (NTG: 33%, Rec. G.: 12.5%, Tox. G.: 33%, Ca. thyroid: 25%). Statistical analysis, however, of results of different patient groups compared to the normal control group failed to show any significance. PMID- 2573627 TI - Irradiation-induced duodenal ulcer disease refractory to ranitidine: healing by omeprazole. AB - In two patients duodenal ulcer refractory to high dose H2-blocker treatment started several months after irradiation therapy following right nephrectomy because of renal adenocarcinoma. Established antiulcer drugs like ranitidine, famotidine, sucralfate, pirenzepine, and antacids alone or in combination were unable to control ulcer pain and failed to induce ulcer healing. Initiation of omeprazole treatment at dosages to produce complete achlorhydria were necessary for ulcer healing and maintenance therapy. We suggest that (a) irradiation may cause duodenal ulcer disease indistinguishable from idiopathic duodenal ulcer; (b) radiation-induced ulcers in the duodenal bulb are refractory to various antiulcer drugs but may heal after administration of omeprazole in dosages that completely suppress acid secretion. PMID- 2573628 TI - Azodisalicylate (Dipentum)-induced hepatitis? PMID- 2573629 TI - Sulfasalazine-induced impotence: a beneficial resolution with olsalazine? PMID- 2573630 TI - Combined use of immunoperoxidase and radioimmunocytochemistry for double immunocytochemical labeling of neurons at light and electron microscopic level. AB - Complexes formed by binding 125I- or 3H-labeled neuropeptides to one of the two binding sites of their specific antibodies allowed specific and sensitive labeling of various peptidergic neurons, which could be detected by classical autoradiographic methods. To visualize two neuronal antigens on the same material at both light and electron microscopic level, we used a new technique of double immunocytochemical labeling, combining immunoperoxidase and radioimmunocytochemistry. The main steps of the process included: (a) indirect labeling of the first antigen by its specific antibody and by a peroxidase labeled Fab immunoglobulin fragment directed against the primary antibody; (b) direct labeling of the second antigen by a radiolabeled peptide-antibody complex; (c) revealing of the first label in the presence of peroxidase substrate; and (d) revealing of the second label by autoradiographic treatment of tissue sections. Compared with other known techniques of double immunostaining, this technique offers major advantages for combined visualization of two neuronal antigens at the electron microscopic level: (a) two neuron types can be labeled by a pre embedding approach, allowing highly sensitive detection of neuronal antigens throughout the 50-microns thickness of vibratome sections; (b) two primary antibodies obtained in the same species can be used to label the two antigens without any risk of crossreactions between the two successive labelings; and (c) the two labels can easily be differentiated, even when they are co-localized within the same neuron structures. Application of this double immunostaining technique is illustrated by data obtained in rat hypothalamus concerning the relationships among a variety of identified neurons and the co-localization of different neuropeptides within the same neuron system. PMID- 2573631 TI - Distribution of adenosine deaminase complexing protein (ADCP) in human tissues. AB - The normal distribution of adenosine deaminase complexing protein (ADCP) in the human body was investigated quantitatively by ADCP-specific radioimmunoassay (RIA) and qualitatively by immunohistochemistry. In these studies we used a specific rabbit anti-human ADCP antiserum. In all 19 investigated tissues, except erythrocytes, ADCP was found by RIA in the soluble and membrane fractions. From all tissues the membrane fractions contained more ADCP (expressed per mg protein) than the soluble fractions. High membrane ADCP concentrations were found in skin, renal cortex, gastrointestinal tract, and prostate. Immunoperoxidase staining confirmed the predominant membrane-associated localization of the protein. In serous sweat glands, convoluted tubules of renal cortex, bile canaliculi, gastrointestinal tract, lung, pancreas, prostate gland, salivary gland, gallbladder, mammary gland, and uterus, ADCP immunoreactivity was found confined to the luminal membranes of the epithelial cells. These data demonstrate that ADCP is present predominantly in exocrine glands and absorptive epithelia. The localization of ADCP at the secretory or absorptive apex of the cells suggests that the function of ADCP is related to the secretory and/or absorptive process. PMID- 2573632 TI - Immunoregulatory function of CD3+, CD4-, and CD8- T cells. Gamma delta T cell receptor-positive T cells from nude mice abrogate oral tolerance. AB - Previous work has shown that abrogation of oral tolerance is mediated by T cells which are found in the CD3+, L3T4- (CD4-), and Lyt-2- (CD8-) subset (termed double-negative; DN) in mice. Inasmuch as it is known that athymic, nude (nu/nu) mice possess Thy 1+, CD4-, and CD8- T cells which also exhibit a functionally rearranged TCR gamma-chain, we investigated whether this subset of nude T cells contained functional immunoregulatory cells. In this report, we examined the phenotype and distribution of CD3+ T cells in the spleen and in the mesenteric and peripheral lymph nodes of BALB/c nu/nu mice in comparison with normal mice (+/+). In the spleens of nude mice, the predominant CD3+ T cell subpopulation was DN. Further, in mesenteric and peripheral lymph nodes, approximately one-third and one-half of the CD3+ T cells were double negative, respectively. In contrast, CD3+, DN T cells represent a small subpopulation in normal (+/+) mice. We next showed that functional regulatory T cells which possess the ability to abrogate oral tolerance were induced in nu/nu mice by Ag priming. BALB/c nude mice were immunized with SRBC, and the splenic CD3+, Vicia villosa-adherent cells were obtained by panning. Adoptive transfer of CD3+, V. villosa-adherent T cells to orally tolerant BALB/c mice restored responsiveness to SRBC, whereas V. villosa nonadherent cells were without effect. In other experiments, CD3+ T cells from the spleens of SRBC-primed mice were further enriched for the CD5+, DN phenotype and adoptive transfer of this subset completely abrogated oral tolerance to SRBC. To characterize the nature of the TCR expressed on these CD3+, DN T cells, we developed a rabbit antibody to a synthetic peptide (residues 209-218: Tyr-Ala-Asn Ser-Phe-Asn-Asn-Glu-Lys-Leu) which was synthesized from a deduced sequence of the murine delta-gene. Immunoprecipitation of a cell membrane fraction from CD3+, DN T cells with anti-delta TCR antibody isolated a 45-kDa band. Furthermore, immunoprecipitation of these cells with anti-CD3 (145-2C11) revealed bands at 45 and 35 kDa (corresponding to delta- and gamma-chains, respectively). Taken together, these results are the first to show that gamma delta-TCR bearing CD3+, CD4-, and CD8- T cells are functional and reverse oral tolerance when adoptively transferred. PMID- 2573633 TI - Suppression of antibody synthesis by CD4+ T cell clones and normal T cells stimulated with monoclonal anti-CD3 antibody. AB - CD4+ve Th1 clones, as well as normal splenic T cells, were found to suppress LPS driven antibody secretion in a non-Ag-specific and non-MHC-restricted manner when the T cells were activated with the anti-CD3 mAb, 145-2C11. Suppression was observed with both primed and naive B cells, as well as with purified hapten specific B cells, a result that suggests a direct effect of anti-CD3-activated T cells on B cell differentiation. Th1 clones activated by cognate Ag also suppressed LPS-driven antibody secretion. Furthermore, suppression of LPS-driven antibody secretion could be achieved across a cell-impermeable porous membrane when T cells were activated with anti-CD3. Suppression by Th1 clones and by normal T cells could not be attributed to a concomitant decrease in B cell proliferation or to a shift in the kinetics or isotype of the antibody response. These data demonstrate that CD4+ve Th1 clones, as well as normal T cells, can effect suppression of polyclonal antibody formation. PMID- 2573635 TI - CD4+ suppressor cells differentially affect the production of IFN-gamma by effector cells of experimental autoimmune encephalomyelitis. AB - Spleen cells from rats that have recovered from experimental autoimmune encephalomyelitis (EAE) suppress the production of IFN-gamma by effector T cells of EAE in an Ag-specific manner. These postrecovery suppressor cells also inhibit EAE in vivo. Fractionation of the postrecovery suppressor spleen cells on nylon wool and OX-8 coated plates yields a nylon wool-adherent CD4+ suppressor cell population that, when cocultured with effector T cells, suppresses IFN-gamma production by these effector cells. In contrast, the nylon wool-adherent, CD4+ postrecovery suppressor cell population fails to inhibit the production of IL-2 by the effector T cells. In further experiments, the effector T cell population was depleted of CD8+ cells and cocultured with the nylon wool-adherent, CD4+ postrecovery suppressor cells, and the supernatants were assayed for IFN-gamma and IL-2. IFN-gamma production was inhibited in these cultures but IL-2 production was not inhibited. Irradiated effector T cells were cocultured with CD4+ postrecovery suppressor cells, without myelin basic protein, in an effort to determine whether the mechanism of differential lymphokine suppression involved an anti-idiotypic response against effector T cells. No IL-2 was produced, indicating that there was no CD4+ suppressor cell mediated anti-idiotypic response against effector T cells. These studies suggest that the suppressor cell is a nylon wool adherent, CD4+ T cell that functions to down-regulate EAE effector T cells by differential inhibition of lymphokine production. PMID- 2573634 TI - Specific inhibition of hybrid resistance in F1 hybrid mice pretreated with parent strain spleen cells. III. Study of the mechanism. AB - The rejection of H-2b parental bone marrow graft by lethally irradiated F1 recipients, that is known as hybrid resistance (HR), is a multistep process. In a first step a 5-fluorouracil (5-FU)-sensitive T cell recognizes the parental bone marrow cells and stimulates a macrophage-like cell to secrete IFN-alpha/beta (recognition phase). IFN-alpha/beta in turn activates a cyclophosphamide sensitive NK-like cell that is the effector cell for HR (effector phase). In a previous paper we described that HR is specifically abrogated by the pretreatment of the F1 recipient with H-2b parental spleen cells. This abrogation is due to a Thy-1+CD5+CD4+CD8- nylon adherent suppressor cell of F1 origin. The aim of the present work was to study during which of the different phases of HR the activity of the suppressor cell is exerted. Our results showed that abrogation of HR in (C57BL/6 x C3H)F1 (B6C3F1) hybrids pretreated with B6 spleen cells results from: 1) the suppression of the 5-FU-sensitive T cell; 2) the suppression of the cyclophosphamide-sensitive NK-like cell; and 3) the disappearance of a humoral factor that is present in the serum of normal B6C3F1 hybrids and which seems to be involved in the effector phase of HR. The 5-FU-sensitive T cell is the only target of Thy-1+CD5+CD4+CD8- suppressor cell. The mechanisms responsible for the suppression of the NK-like effector cell and the disappearance of the humoral factor are discussed. PMID- 2573636 TI - Phorbol esters regulate CD2- and CD3-mediated calcium responses in peripheral blood-derived human T cells. AB - The purpose of the present study was to examine the effect of protein kinase C (pkC) activation on calcium responses generated through the CD3 and CD2 Ag in both normal peripheral blood-derived T lymphocytes and the leukemic T cell line Jurkat. The data reveal a major difference with respect to the regulation of receptor-mediated calcium responses in these two cells. Thus, the pkC activator phorbol-12,13-dibutyrate (Pdbu) enhances calcium responses induced via CD3 and CD2 molecules in normal T cells by accelerating the rate of elevation of intracellular calcium levels and increasing the maximum change in calcium concentration achieved. In contrast, Pdbu inhibits both CD3- and CD2-induced calcium responses in Jurkat cells. Pdbu does not influence calcium responses generated by the guanine nucleotide-binding protein activator, aluminium fluoride, indicating that the effect of pkC occurs at a point proximal to a guanine nucleotide-binding protein regulation of T cell calcium responses. PMID- 2573637 TI - Cimetidine and the immuno-response in healthy volunteers. AB - We compared the immunologic measurements from treatment of 12 healthy volunteers (six male, six female) with 800 and 1,600 mg cimetidine. In the first trial 800 mg cimetidine was administered daily to the volunteers over a period of 7 d; after an interruption of 2 months, 1,600 mg of cimetidine was applied daily for 21 d. The most striking result of our study was an increased mitogen-induced lymphocyte proliferation. This conclusion can be drawn from the fact that phytohaemagglutinin (PHA) (0.4 microgram/well) and pokeweed mitogen (PWM) (0.4 microgram/well) induced lymphocyte proliferation were found to be significantly increased in comparison to pretreatment values on day 7 in both cimetidine regimens (800 mg; PHA: mean proliferation 66,500 before treatment to 166,00 cpm, PWM: mean proliferation 8,800 before treatment to 34,000 cpm; 1,600 mg; PHA; mean proliferation 48,700 before treatment to 81,600 cpm; PWM: mean proliferation 6,300 before treatment to 16,200 cpm). Increased mitogen-induced proliferation following cimetidine intake is of special interest because the mechanisms of this activation process are incompletely known. Lymphocyte proliferation response is dependent on the availability of extracellular calcium. The function of the other bivalent cations is unknown. We found that the extent of mitogen-induced lymphocyte proliferation correlates with cellular intralymphocytic zinc and magnesium amounts (coefficients of correlation [r]) (800 mg: PHA/Mg r = 0.84; PHA/Zn r = 0.86; PWM/Mg r = 0.88; PWM/Zn r = 0.87). Though the application of both cimetidine doses causes enhanced mitogen-induced lymphocyte proliferation on day 7, T lymphocytes with different phenotypic properties appear to be influenced by cimetidine. In the first dose regimen (800 mg) the number of the CD8 lymphocytes decreased significantly from 16.1% (365 cell/microliters blood) to 12.7% (264 cells/microliters blood) after 7 d of cimetidine intake. After the period of high-dose (1,600 mg) cimetidine administration (at day 21) numbers of CD4 lymphocytes were significantly increased from 41.5% (860 cells/microliters blood) to 56.3% (1,210 cells/microliters blood). Our results show that although different cimetidine doses obviously influence different cell types of healthy volunteers, the cellular mechanisms are the same, namely, a proliferation and an increased incorporation of magnesium and zinc in lymphocytes. PMID- 2573638 TI - [Diffuse narrowing of the internal mammary artery graft--the thinning phenomenon]. AB - Diffuse narrowing of the internal mammary artery graft (IMAG), the thinning phenomenon, was found in 8 (6.7%) out of 120 IMAGs underwent postoperative angiography. There were 6 males and 2 females, and mean age was 55.3 ranged from 37 to 70 year-old. There was no significant correlation with hyperlipidemia, diabetes mellitus or hypertension. Grafted coronary arteries were 6 anterior descending, one diagonal and one circumflex arteries. Diameter of them was 1.5 mm in one and 2 mm in 7. severity of stenosis of proximal coronary artery was 100% in one, 90-99% in one and less than 90% in 6. Undivided sizable costal or pericardial branches were found in 4 patients. All eight patients were alive without angina, although mild positive stress EKG changes were noted in two patients. In conclusion, a high resistance state such as a good competitive native coronary flow was thought to be the most important factor for the diffuse narrowing of IMAG, and close follow up should be needed because its outcome is still unknown. PMID- 2573639 TI - [Biosynthesis of kyotorphin, a dipeptidic neuromodulator]. PMID- 2573640 TI - [Diagnostic value of serological tumor marker tests in patients with ovarian cancer]. AB - The usefulness of tumor markers in serodiagnosis of cancer designed to detect ovarian cancer at an early stage was evaluated from the point of view of their diagnostic value. Namely, eight tumor markers, CA125, SLX, CA72-4, TPA, Fr, CEA, CA19-9, and SCC, were determined and studied to find the combination that would yield the optimal diagnostic value. For this purpose, the diagnostic value was calculated from sensitivity x specificity. As a single tumor marker CA125 proved optimal with a diagnostic value of 0.50. The higher value, 0.53, was obtained as the diagnostic value from the combination of two tumor markers, CA125 and CA72-4. When three tumor markers were combined, CA125, CA72-4 and SLX gave an optimal diagnostic value of 0.65. In the combination of four tumor markers, CA125, CA72 4, SLX and CA19-9 gave a diagnostic value of 0.63. In the five marker combination CA125, CA72-4, SLX, CA19-9 and TPA worked well and had a diagnostic value of 0.59. When the markers were increased to six types, CA125, CA72-4, SLX, CA19-9, TPA and Fr provided a combination with 0.53 as the diagnostic value. In the seven marker combination CA125, CA72-4, SLX, CA19-9, TPA, Fr and CEA gave a diagnostic value of 0.51. The efficiency declined to 0.51 when eight tumor markers were combined. When cost performance in the measurement of tumor markers for early detection of ovarian cancer is taken into account, a dilemma arises in that the increase in the number of tumor markers used is accompanied by higher sensitivity and lower specificity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573641 TI - The radiobiology of human cells and tissues. In vitro radiosensitivity. The picture has changed in the 1980s. AB - Substantial developments have been made during the 1980s in the radiobiology of human tumours, in particular in studies of the radiosensitivity of human tumour cells. It is now clear that tumour cells differ considerably in radiosensitivity, to an extent that by itself is capable of explaining the clinical response of tumours to radiotherapy. There also is evidence that the radiosensitivity of human tumour cell lines to low radiation doses correlates with clinical experience. Irradiation at low dose rate amplifies the differences between cell lines. In conjunction with mathematical modelling, a study of the dose-rate effect also allows a distinction to be drawn between repairable and non repairable damage. The differences seen between cell lines at low acute doses or low dose rates are associated with the non-repairable component. The most radiosensitive cell lines have a steep component of non-repairable damage and they give the impression of being recovery-deficient; this may, however, be incorrect for when evaluated at constant dose levels recovery is found to increase with increasing radiosensitivity. This leads to the view that recovery from radiation damage may reflect the amount of recoverable damage inflicted rather than the 'capacity' of the cells to recover. PMID- 2573642 TI - The radiobiology of human cells and tissues. Proceedings of the 15th L.H. Gray Conference. University of Kent, Canterbury, U.K., 11-15 April 1989. PMID- 2573643 TI - The relevance of alpha/beta ratios determined in vitro for human cell lines to the understanding of in vivo values. PMID- 2573644 TI - Initial damage or repair as the major determinant of cellular radiosensitivity? PMID- 2573645 TI - Multicellular spheroids from human soft tissue sarcomas: radiocurability and dose fractionation effect. PMID- 2573646 TI - Radiobiological characterization of 53 human tumor cell lines. AB - We investigated the in vitro radiobiological survival parameters of 53 human tumor cell lines studied in exponential growth. Epithelial cell lines derived from 24 patients with head and neck carcinoma, 15 patients with ovarian carcinoma, and mesenchymal tumor cell lines derived from 14 patients with bone and soft tissue sarcomas were studied. Survival data are analyzed using the multi target and linear quadratic models. The head and neck and ovarian carcinoma tumor cell lines were more radioresistant as measured by D0, D, and alpha parameters, compared with the bone and soft-tissue sarcoma lines. The radiobiological parameters of tumor cell lines reported herein are similar to parameters of tumor cell lines derived from head and neck and soft tissue sarcoma patients presently being followed for clinical outcome following radiotherapy in our clinic. PMID- 2573648 TI - Do in vitro studies of potential lethal damage repair predict for in situ results? PMID- 2573647 TI - Radiosensitivity and PLDR in primary cultures of human normal and malignant mammary and prostate cells. PMID- 2573649 TI - Radiation biology of human tumour xenografts. AB - The radiation response of human tumour xenografts can be measured with sufficient accuracy using cell survival in vitro and tumour growth delay in vivo as endpoints. There is evidence that the radiation response of xenografts mirrors the clinical radioresponsiveness of the corresponding tumours in patients. Consequently, xenografts may have a significant potential in experimental radiotherapeutic research, e.g. in the development of in vitro and in vivo predictive assays of clinical radioresponsiveness. However, there are at least three main disadvantages with xenografts as models for human cancer. Firstly, the volume doubling time is usually shorter for xenografts than for tumours in patients. Secondly, the haematological system and the vascular network of xenografts originate from the host. Thirdly, host defence mechanisms may be active against xenografts. These disadvantages may limit the usefulness of xenografts as models for human cancer in some types of radiobiological studies. PMID- 2573650 TI - Comparison of the radiobiological properties of human tumour xenografts and rodent tumours. PMID- 2573651 TI - Manipulation of radiobiological hypoxia in a human melanoma xenograft to exploit the bioreductive cytotoxicity of RSU-1069. PMID- 2573652 TI - Radiosensitivity and repair capacity of two xenografted human soft tissue sarcomas to photons and fast neutrons. PMID- 2573653 TI - The relationship between potentially lethal damage repair and intrinsic radiosensitivity of human cells. AB - The intrinsic radiosensitivity of exponentially growing cells (exp) was compared to that of immediately plated plateau phase cells (ip) using published data on 60 human cell lines (27 fibroblast lines and 33 tumour cell lines). The values for alpha, D and S2 are not significantly different for the two groups; beta is significantly higher in ip cells. This produces a smaller alpha/beta ratio in ip cells than in exp cells. The influence of potentially lethal damage (PLD) repair was assessed by comparing the radiosensitivities of ip cells and plateau phase cells with delayed plating (dp). The published data for 81 human cell lines (48 fibroblasts and 33 tumour lines) were used. PLD repair was found to lead to a decrease in alpha and an increase in D and S2, whereas neither beta nor the alpha/beta ratio changed significantly. The relationship between PLD repair and intrinsic radiosensitivity was assessed by repair capacity and the repair ratio. The fitted relationship is a bell-shaped curve with a maximum at 2.2 Gy for repair capacity. The fitted curve predicts that repair capacity is zero at a D up of 0.28 Gy and at 4 Gy. Thus, PLD repair is a reasonable reflection of intrinsic radiosensitivity up to 2.2 Gy. Above 2.2 Gy, the relationship is reversed: the greater the radioresistance, the lower the PLD repair. PMID- 2573654 TI - Radiation studies on sensitivity and repair of human mammary epithelial cells. PMID- 2573655 TI - Radiobiological heterogeneity of leukemic lymphocyte precursors from acute lymphoblastic leukemia patients. PMID- 2573656 TI - The radiosensitivity of human haemopoietic progenitor cells. PMID- 2573657 TI - The initial physical damage produced by ionizing radiations. AB - Biophysical studies of different ionizing radiations and their differences in biological effect can provide useful information and constraints on the nature of the initial biologically relevant damage and hence the subsequent biochemistry and repair processes. It is clear that the nature of the predominant critical component produced by densely ionizing (high-LET) radiations is qualitatively, as well as quantitatively, different from that which predominates for low-LET radiations. Comparisons of radiation track structure with observed biological effects of the radiations allow hypotheses to be developed as to the nature of these different types of damage. That associated with low-LET radiations seems consistent with what is known about DNA double-strand breaks (dsb). It is produced predominantly by a localized cluster of ionizations within a single electron 'track end' either by direct action on the DNA or in conjunction with closely-associated molecules. The characteristic high-LET damage is somewhat larger in number of ionizations and spatial extent and therefore presumably also in molecular complexity. It is suggested that the total spectrum of initial damage be categorized into four classes; in addition to the above two this would include on the one extreme sparse isolated ionizations, which may lead to very simple products that are of limited biological relevance, and on the other extreme very large and relatively rare events which are uniquely achievable by some high-LET radiations, such as alpha-particles, but not at all by low-LET radiations. These biophysical considerations pose a challenge to radiation chemistry studies to consider the chemical consequences of highly localized clusters of initial ionizations and excitations in or very near to DNA, and to biochemistry to consider classes of damage involving DNA (and perhaps associated molecules) of greater complexity than the simplest dsb. PMID- 2573658 TI - On the nature of interactions leading to radiation-induced chromosomal exchange. AB - Within the conceptual framework of so-called lesion-interaction models, chromosomal interchanges are believed to result from radiation damage to both chromosomes involved. More recently, models of radiation action have been proposed which suggest such exchanges arise from initial damage to only one chromosome, which then associates with an undamaged chromosome. The specific case of 'lesion-nonlesion' chromosomal interaction via telomere-break rejoining was examined through the use of a telomere-specific DNA probe. No evidence was found to support dicentric formation by this mechanism in normal human fibroblasts. To test the more general case (i.e. lesion-nonlesion interaction by some other mechanism) mitotic HeLa cells were fused together to determine whether exchanges would occur between the chromosomes of previously separate genomes, as seen in resulting cell syncytia at the next mitosis. The fusion of irradiated cells (with each other) produced a high frequency of such intergenomic exchanges. However, the frequency of these events was reduced 50-100-fold in syncytia resulting from the fusion of irradiated with unirradiated cells. These results strongly support the view that most radiation-induced exchange aberrations require initial damage to chromatin at both locations involved in the exchange--i.e. they are fundamentally two-hit in nature. PMID- 2573659 TI - Induction and rejoining of chromatid breaks in X-irradiated A-T and normal human G2 fibroblasts. PMID- 2573660 TI - The radiosensitivity of human neuroblastoma: a cellular and molecular study. PMID- 2573661 TI - Cell cycle responses of two X-ray sensitive mutants defective in DNA repair. AB - Both the xrs and V-3 lines of Chinese hamster ovary cells exhibit marked sensitivity to ionizing radiation. They are also sensitive to agents such as bleomycin and H2O2 but exhibit normal responses to ultraviolet light and mitomycin C. Both cell lines are defective in split-dose repair and repair of double-strand breaks in DNA. Analysis of response to radiation as a function of age in the cell cycle indicates that both cell lines exhibit a marked sensitivity in late G1 and early S phase with more limited sensitization throughout the remainder of the cell cycle. PMID- 2573663 TI - Increased radiosensitivity and the basic defect in ataxia telangiectasia. AB - Various cellular defects have been found in ataxia telangiectasia (A-T) cells including increased radiosensitivity, increased sensitivity to various chemical agents, a probable DNA repair defect and a defect in DNA synthesis. How these different features are related to each other is at present unknown. It has been suggested that there is a defect in A-T that acts in tissue differentiation as well as during growth and in the mature adult. This hypothesis is supported by the observations, for example, of an immature thymus present in patients, the production of alpha-fetoprotein, which results in a high serum level, and ovarian dysgenesis. A gene for A-T has recently been localized to chromosome region 11q22 23, a site involved in chromosomes translocations in some non-lymphoid leukaemias. At the chromosomal level the spontaneous abnormalities in A-T include, first, an increased frequency of cells showing chromosome translocations involving immune system genes that normally undergo rearrangement to form a functional product; secondly, the formation of telometric dicentrics in both lymphocytes and fibroblasts; and thirdly formation of long-lived chromosome damage following exposure to ionizing radiation and radiomimetic drugs. The gene defect underlying this disorder is unknown and distinguishing between primary and secondary effects of the mutant gene is difficult. We consider alternative models for retention of translocation T cells. First, it is possible that there is a defect in recognition of site-specific damage leading to retention of translocation cells that might otherwise be removed. Secondly, a feature common to the production of illegitimate T-cell receptor gene rearrangements and to formation of telomeric dicentric chromosomes in A-T cells is an increased period of time available for chromosome interchange, possibly due to a site-specific defect in strand break repair. It is possible that this defect may also prevent chromosome restitution following exposure of cells to ionizing radiation. PMID- 2573662 TI - Stable radioresistance in ataxia-telangiectasia cells containing DNA from normal human cells. AB - SV40-transformed ataxia-telangiectasia (AT) cells were transfected with a cosmid that contains a normal human DNA library and a selectable marker, the neo gene, which endows successfully transformed mammalian cells with resistance to the antibiotic G418. After a three-part selection protocol for G418 resistance and radioresistance, a cell line stably resistant to ionizing radiation was recovered. Cells from this line were irradiated with 50 Gy of X-rays and fused with non-transfected AT cells. Among the G418-resistant colonies recovered was one that was stably resistant to radiation. Resistance to ionizing radiation of both the primary transfectant line and its fusion derivative was intermediate between that of AT cells and normal cells, as assayed by colony-forming ability and measurement of radiation-induced G2 chromatid aberrations; both cell lines retained AT-like radioresistant DNA synthesis. These results suggest that, because radioresistance in the transfected cells was not as great as that in normal human cells, the two hallmarks of AT, radiosensitivity and radioresistant DNA synthesis, may still be the result of a single defective AT gene. PMID- 2573664 TI - Studies of DNA damage and repair of thermal- and radiation-induced lesions in human cells. PMID- 2573665 TI - Response of human organs to single (or fractionated equivalent) doses of irradiation. AB - Characteristics of the kinetics of radiation response of human tissues and organs are exemplified by effects in the testis and the ovary. Also, published dose incidence curves for specified levels of injury in bone marrow, liver, bladder and lung are characterised in terms of single doses as well as single-dose equivalents calculated from fractionated doses using the alpha/beta equation. It is shown that these curves, analysed using a Poisson model, have slopes characterised by D0-equivalents ranging between 1.25 and 2.5 Gy. These values are higher or within the range of values reported in general for single-dose survival curves of human cells in primary culture (range of D0 values 0.7-1.8 Gy). This indicates that single-cell responses together with other complicating biological and statistical sources of heterogeneity under discussion, could form a basis for explaining the steepness of dose-incidence curves for organ injury after fractionated doses. With local organ irradiation, increase in the single-dose equivalent by 3-10 per cent would increase the complication rate from 5 per cent to 10 per cent. Higher dosage increases (by up to two times) apply to fractionated doses. PMID- 2573666 TI - Fractionation parameters for human tissues and tumors. AB - Time-dose factors such as fractionation sensitivity (alpha/beta) can sometimes be estimated from clinical data, when there is a wide variation in dose, fraction size, treatment time, etc. This report summarizes estimates of fractionation parameters derived from clinical results. Consistent with the animal data, alpha/beta is higher for acutely responding than for late-responding normal tissues. While many human tumors seem to be characterized by high alpha/beta values, there are exceptions (e.g. melanomas). Repair kinetics may be slower in human than in rodent skin and mucosa, but there are no hard and fast estimates of the repair halftime. Regeneration in head and neck tumors is equivalent to a daily dose of 1 Gy or less, while in the mucosa it is equivalent to approximately 1.8 Gy/day. PMID- 2573668 TI - A comparison of the normal-tissue reactions in patients treated with either 3F/Wk or 5F/Wk in the BIR (British Institute of Radiology) trial of radiotherapy for carcinoma of the laryngo-pharynx. PMID- 2573667 TI - Early and late normal-tissue injury after postmastectomy radiotherapy alone or combined with chemotherapy. PMID- 2573669 TI - Does smoking protect against radiation-induced pneumonitis? PMID- 2573670 TI - Clinical interest in determinations of cellular radiation sensitivity. AB - Determinations of cell sensitivity in terms of survival fraction after doses employed in clinical radiation therapy, say 1-3 Gy, are of increasing interest to clinicians as they provide direct experimental data which can be employed without reference to models of cell inactivation. SF2 values are expected ultimately to prove valuable as response predictors. Even so, SF2 values would surely be combined with other predictors also under development to give the best feasible estimate of response of tumor and normal tissue. There are, however, several concerns with the SF2 data currently available. These include: SF2 depends upon the cell system employed (established cell lines vs primary cultures) and the method of assaying survival fraction (colony formation vs population growth); dose-response curves for inactivation of tumors characterized by the reported distribution of SF2 values are, in many instances, not close to those judged to obtain in clinical practice; the broad distribution of SF2 values indicates a rather flatter dose-response curve for tumor control or normal tissue than seems true from clinical experience. There appears to be a potential for clinical gain by determination of sensitivity of normal tissues in order to identify patients who are of increased sensitivity (for example heterozygotes for AT, 5 oxoprolinuria, etc.). Although the absolute SF2 values obtained by current technologies of culturing human cells often appear to be poorly related to values expected from observed radiation response in patients, intensive research on cell viability assays will almost certainly yield more realistic results. PMID- 2573671 TI - Expected changes in local tumor control rates resulting from the selective use of accelerated fractionation. PMID- 2573672 TI - Dose-rate effects in biologically targeted radiotherapy. PMID- 2573673 TI - Radiosensitivity of human head and neck squamous cell carcinomas in primary culture and its potential as a predictive assay of tumor radiocurability. AB - The intrinsic radiosensitivity of human tumor cell cultures correlates with the clinical radiosensitivity of several different tumor histologies, as evidenced by analyses of low dose parameters of radiation survival curves generated from a large number of cell lines. Such radiosensitivity has therefore served as a basis of attempts to develop predictive assays of tumor radiocurability. In this study, the tumors from 72 patients with head and neck squamous carcinoma have been grown in an adhesive tumor-cell assay system and radiosensitivity (S2: survival at 2.0 Gy) values have been measured. The characteristics of these cultures, such as growth rate, clonogenicity and growth enhancement by epidermal growth factor, do not correlate with S2. The average S2 value of the 72 cultures is 0.33, which is lower than for cultures derived from melanomas and lung adenocarcinomas. Twenty six patients followed up for at least 15 months have been evaluated for local tumor control. The average S2 value of the seven patients with recurrences in this group is slightly higher (0.43) than that from the other patients (0.30). There is considerable overlap of S2 values in the two groups, and more patients must be evaluated before the groups can be compared statistically. PMID- 2573674 TI - Evaluation of surviving fraction at 2 Gy as a potential prognostic factor for the radiotherapy of carcinoma of the cervix. PMID- 2573675 TI - Linear correlation between surviving fraction and the micronucleus frequency. PMID- 2573676 TI - Can cell kinetic parameters predict the response of tumours to radiotherapy? AB - Three potential predictive assays of the repopulation component in tumour response to therapy are considered. (1) The DNA index can easily be measured. It is of prognostic value for cancers of certain sites, aneuploidy being a bad prognostic indicator. It is not strictly an indicator of cell proliferation. (2) The in vitro labelling index is of predictive value in early stage operable breast cancer and in head and neck cancer. In the former a high pretreatment labelling index can identify patients who could benefit from adjuvant chemotherapy. (3) The tumour potential doubling time (Tpot) can be measured rapidly following in vivo labelling with bromodeoxyuridine or iododeoxyuridine. We have measured Tpot in over 100 solid tumours with a success rate of about 75 per cent. Nearly 50 per cent of the tumours have a pre-treatment potential doubling time of 5 days or less. These would be suitable candidates for accelerated fractionation. PMID- 2573677 TI - In vivo bromodeoxyuridine labelling in human tumour xenografts. PMID- 2573678 TI - Ki67 in the assessment of tumour growth rate: a study on xenografts. PMID- 2573679 TI - Sensitization of hypoxic tumour cells--clinical experience. AB - There is substantial evidence for the presence of hypoxia in human tumours. This is documented by histopathological demonstration of vascular insufficiency, direct oxygen measurements in tumours, as well as by physiological imaging and mapping of hypoxic areas. As a consequence, clinical trials have focused on the hypoxia problem for more than 30 years. This includes the use of hyperbaric oxygen, hypoxic cell radiosensitizers, and, more recently, modification of the oxygen-unloading capacity of haemoglobin. Agents directed towards destruction of hypoxic cells have also been applied, such as hyperthermia and bio-reductive drugs. Despite decades of clinical trials, the results are still inconclusive, and although some trials have shown significant benefit, it has become apparent that hypoxia is a complex problem. Hypoxia appears to be especially a problem in certain tumour types (e.g. squamous cell carcinoma), but even within tumours of the same type, site, and stage, hypoxia does not occur to the same extent. Furthermore, there are increasing suggestions that hypoxia may occur in two principally different ways, namely acutely and chronically, yielding varying responses to modifying agents. Although improvement in hypoxic cell radiosensitizers and other agents is under way, a definitive solution to the hypoxia problem will not be found until the tumours in which hypoxia occurs can be identified. This will require detailed analysis of individual tumours and patients' parameters, and better knowledge of the mechanisms of reoxygenation in clonogenic tumour cells. PMID- 2573680 TI - Comparison between X-rays and SR 4233 for cytotoxicity and repair of potentially lethal damage in human cells. PMID- 2573681 TI - A comparison of thermal responses of human and rodent cells. AB - A comparison of heat responses of cells from human and rodent tumors indicates the following: (1) human cells appear to be appreciably more heat-resistant than are rodent cells in the range 41-45 degrees C; (2) while rodent cells show a marked increase in sensitivity as the temperature is increased from 42 and 43 degrees C, this change occurs at approximately 44 degrees C or higher for the human lines examined; (3) rodent cells are unable to acquire thermotolerance during exposure to 43 degrees C; the human cells do so readily; (4) decay of tolerance tends to be complete in 72 h in rodent cells; in human cells it may take twice that time. These results may have important implications for the clinical use of hyperthermia. PMID- 2573682 TI - Radiobiology and clinical application of halogenated pyrimidine radiosensitizers. AB - Halogenated pyrimidines (HP) represent a unique class of non-hypoxic cell radiosensitizers currently under clinical re-investigation. In order for halogenated pyrimidines to sensitize cells to radiation, they must be incorporated into cellular DNA. In the case of human tumors, which have in general rather long cell cycle times, this may require many days of continuous drug infusion to achieve adequate replacement of the DNA base thymidine with HP. In vitro studies support the relationship between the extent of radiosensitization and the percentage of thymidine replacement. Recent clinical studies evaluating the role of iododeoxyuridine (IdUrd) as a radiation sensitizer in large unresectable sarcomas have been extremely encouraging. To support and expand upon these positive clinical findings more information and research is needed regarding: (1) the mechanism of HP-induced radiosensitization; (2) the percentage of HP thymidine replacement in human tumors achievable and how it relates to treatment outcome; (3) the means of increasing HP incorporation in tumor and minimizing incorporation in normal tissues; (4) a better understanding of optimal timing between HP administration and radiation treatment; and (5) methods to evaluate which tumors are appropriate candidates for HP therapy. While presently limited to use in conventional high dose-rate X-ray therapy, laboratory studies suggest that HP might also be effective in low dose-rate brachytherapy and for selected high LET clinical beams. HPs probably will not be 'general' non hypoxic cell radiosensitizers for all tumor types, but with appropriate tumor type/anatomical site selection and refinement in their administration, HPs may prove beneficial in cancer treatment. PMID- 2573683 TI - Selenite-induced increase in glutathione peroxidase activity protects human cells from hydrogen peroxide-induced DNA damage, but not from damage inflicted by ionizing radiation. PMID- 2573685 TI - Pathogenesis of Hantaan virus in mice. AB - The virulence of two virus clones (HV cl-1 and HV cl-2) of Hantaan virus, which were plaque-purified on Vero E6 cells, were compared in suckling mice infected by the intraperitoneal or intracerebral route. HV cl-1 increased the mortality of the mice whereas HV cl-2 did not. Furthermore, virus of high titre was isolated from various organs of mice infected with HV cl-1 and high titres were maintained, whereas after infection with HV cl-2, virus was isolated from various organs only in low titre and only temporarily. HV cl-1 strongly induced cell-to cell fusion, but the cell fusion activity was at a minimum level in cells infected with HV cl-2. However these two clones induced similar titres of antibodies in mice. Cytotoxic T lymphocyte assays against macrophages infected with homologous and heterologous virus showed that cytotoxic T cell activity was induced in mice infected with HV cl-2, but suppressed in mice infected with HV cl 1. These results suggest that an alteration in the cell fusion function and the cytotoxic T cell activity are important in the pathogenesis of Hantaan virus infection in newborn mice. PMID- 2573686 TI - Restriction fragment differences between the genomes of the Oka varicella vaccine virus and American wild-type varicella-zoster virus. AB - The Oka vaccine strains of varicella-zoster virus (VZV) have a significantly different BgII DNA restriction pattern from that of American wild-type isolates of VZV. This difference consists primarily of an additional BgII site, which lies within the BamHI "D" fragment. In conjunction with a study of the efficacy of an experimental Merck/Oka VZV vaccine, the area of the genome from which the most marked restriction pattern alteration arises was studied more closely to determine if there are other significant differences between the Oka strains and American wild-type strains. BamHI "D" fragments from the DNA of the Oka parent strain (the progenitor of the vaccine strain), the RIT/Oka vaccine strain (a derivative of the Oka parent strain), the Merck/Oka vaccine strain, and the EF strain (an American wild type), were submitted to extensive endonuclease digestion studies to ascertain if additional unique restriction sites are present in the Oka parent or vaccine strains. The extra BgII restriction site characteristic of the Merck/Oka vaccine strain is also present in the DNA of the parent virus as well as its derivatives and was therefore not produced by the "attenuation" process. No other novel sites were found in the Oka parent or Oka derived strains in this section of the genome. The Merck/Oka vaccine strain of VZV, despite its Japanese origin, is therefore quite similar to circulating American varicella-zoster virus strains. Varicella-zoster virus DNA, at least in the area of the BamHI D fragment, also appears to be remarkably stable from strain to strain. PMID- 2573684 TI - MHC control of CD4+ T cell subset activation. AB - The present results demonstrate that CD4+ T cells activated in the primary in vivo response to antigen produce distinct patterns of cytokines depending upon the MHC class II haplotype of the responding mice. I-As mice were found to selectively activate IL-2/IFN-gamma-producing CD4+ T cells, whereas I-Ab mice exhibited selective activation of IL-4-producing CD4+ T cells in response to collagen IV. The effector response phenotype was found to correlate with the cytokine phenotype of CD4+ T cells activated in vivo; IL-2/IFN-gamma-producing cells giving rise to proliferative (cell-mediated) responses, IL-4-producing cells leading to secondary IgG (humoral) responses. Together the data support the notion that the outcome of a given immune response (e.g., protection vs. onset, tolerance vs. autoimmunity) may be determined in part by the type of CD4+ T cells initially activated by antigen. Moreover, the present experiments demonstrate for the first time that polymorphism in class II MHC can determine such selective activation of different cytokine-producing CD4+ T cell phenotypes. PMID- 2573687 TI - Carbon dioxide and 1-octen-3-ol as mosquito attractants. AB - Interval suction traps were used to study the attractant effect of CO2 and 1 octen-3-ol on trap catches of mosquito populations at 2 different locations in Florida. There was no significant increase in the numbers of mosquitoes caught when the concentration of CO2 was increased from 200 to 1,000 cc/min. One-octen-3 ol used by itself attracted mosquitoes in numbers similar to CO2 released at 200 cc/min. One-octen-3-ol and CO2 acted synergistically in attracting significantly greater numbers of Aedes taeniorhynchus, Anopheles spp. and Wyeomyia mitchellii than either bait used singly, although the response of Culex spp. to this bait combination was less pronounced. Ceratopogonidae (Culicoides furens) and Tabanidae (Diachlorus ferrugatus, Tabanus nigrovittatus and Chrysops spp.) were also attracted to the combined bait. PMID- 2573688 TI - Egg cannibalism and carnivory among three species of Toxorhynchites. AB - In laboratory experiments, third and early fourth instar larvae of Toxorhynchites amboinensis, Tx. splendens and Tx. brevipalpis, previously starved 24 h, rapidly cannibalized eggs of their own species, or ate the eggs of other species present on the water surface in small containers. Toxorhynchites amboinensis and Tx. splendens larvae of either instar cannibalized eggs somewhat more rapidly than Tx. brevipalpis, probably because brevipalpis eggs distributed themselves around the edge of the container and were less accessible. When offered heterospecific eggs, fourth instar larvae of all three species ate them as efficiently as they cannibalized their own except that eggs of Tx. brevipalpis were eaten very slowly. Toxorhynchites amboinensis larvae were offered conspecific hatched eggs, and these also were consumed. PMID- 2573689 TI - Conceptual model for the use of aerial color infrared photography by mosquito control districts as a survey technique for Psorophora columbiae oviposition habitats in Texas ricelands. AB - Two photographic missions per year are recommended to provide information on land use and mosquito oviposition habitats. A winter mission, following a rain, will provide a view of low areas within fields which may be obscured by summer vegetation. A summer mission will provide current land-use and crop distribution information and may show plant stress conditions due to excessive soil moisture. An aerial color infrared photographic survey with directed ground verification should result in a substantial savings in cost and increased efficiency in surveillance of mosquito producing habitats over ground survey techniques currently employed by mosquito control districts. PMID- 2573691 TI - Bionomics of Culiseta particeps in southern California. PMID- 2573690 TI - Parasitism of Coquillettidia perturbans by two water mite species (Acari: Arrenuridae) in Florida. AB - Female Coquillettidia perturbans collected in northern Florida were commonly parasitized by 2 species of water mites. Earlier in the year, mosquitoes were parasitized primarily by Arrenurus danbyensis and later in the year primarily by Arrenurus delawarensis. The number of mosquitoes simultaneously parasitized by both parasites is apparently greater than expected due to chance. PMID- 2573692 TI - A modification of scrap automobile tires for field studies of artificial container-breeding mosquitoes. PMID- 2573693 TI - Preliminary field observations on the killing behavior of Toxorhynchites amboinensis larvae. PMID- 2573694 TI - Literature references for mosquitoes and mosquito-borne diseases. 1989--Part 3. PMID- 2573696 TI - GABA as an inhibitory neurotransmitter in human cerebral cortex. AB - 1. The possible role of gamma-aminobutyric acid (GABA) as an inhibitory neurotransmitter in the human cerebral cortex was investigated with the use of intracellular recordings from neocortical slices maintained in vitro. 2. Electrical stimulation of afferents to presumed pyramidal cells resulted in an initial excitatory postsynaptic potential (EPSP) followed by fast and slow inhibitory postsynaptic potentials (IPSPs). The early IPSP had an average reversal potential of -68 mV, was associated with a mean 67-nS increase in membrane conductance, was reduced by the GABAA antagonist bicuculline, was sensitive to the intracellular injection of Cl-, and was mimicked by the GABAA agonist muscimol. 3. The late IPSP, in contrast, had an average reversal potential of -95 mV, was associated with a mean 12-nS increase in membrane conductance, was reduced by the GABAB antagonist phaclofen, and was mimicked by the GABAB agonist baclofen. 4. Block of the early IPSP by bicuculline or picrotoxin led to the generation of paroxysmal epileptiform activity, which could be further enhanced by reduction of the late IPSP. 5. These data strongly support the hypothesis that GABA is a major inhibitory neurotransmitter in the human cerebral cortex and that GABAergic IPSPs play an important role in controlling the excitability and responsiveness of cortical neurons. PMID- 2573695 TI - ALADDIN: an integrated tool for computer-assisted molecular design and pharmacophore recognition from geometric, steric, and substructure searching of three-dimensional molecular structures. AB - ALADDIN is a computer program for the design or recognition of compounds that meet geometric, steric, and substructural criteria. ALADDIN searches a database of three-dimensional structures, marks atoms that meet substructural criteria, evaluates geometric criteria, and prepares a number of files that are input for molecular modification and coordinate generation as well as for molecular graphics. Properties calculated from the three-dimensional structure are described by either properties calculated from the molecule itself or from the molecule as compared to a reference molecule and associated surfaces. ALADDIN was used to design analogues to probe a bioactive conformation of a small molecule and a peptide, to test alternative superposition rules for receptor mapping of the D2 dopamine receptor, to recognize unexpected D2 dopamine agonist activity of existing compounds, and to design compounds to fit a binding site on a protein of known structure. We have found that series designed by ALADDIN show much more subtle variation in shape than do those designed by traditional methods and that compounds can be designed to be very close matches to the objective. PMID- 2573697 TI - N-methyl-D-aspartate antagonists prevent interaction of binocular maps in Xenopus tectum. AB - Glutamate receptors appear to play a key role in several forms of experience dependent modification of both the strength of synapses and synaptic connectivity. In developing Xenopus frogs, the connections made by isthmotectal axons relaying visual input from the eye to the ipsilateral tectum are markedly influenced by the visual activity of contralateral retinotectal axons, and normal binocular visual input is necessary in order for the ipsilateral visuotectal map to come into register with the contralateral map. We have tested whether NMDA receptors play a role in establishment of the topographic matching of binocular maps during development. We have examined the effects of chronic treatment of tectum with either the receptor agonist NMDA or the antagonists APV or CPP applied throughout early postmetamorphic life using subpial implants of drug impregnated elvax. Both antagonists blocked the matching of the ipsilateral map to the contralateral map, while NMDA permitted such matching. Our data therefore indicate that NMDA receptors are involved in the experience-dependent establishment of matching binocular maps during development. PMID- 2573698 TI - Disappearance and reformation of synaptic vesicle membrane upon transmitter release observed under reversible blockage of membrane retrieval. AB - The temperature-sensitive mutant of Drosophila, shibire(ts-1), which is normal at 19 degrees C, but in which endocytosis is reversibly blocked at 29 degrees C, was used to deplete synapses of vesicles by inducing transmitter release while membrane retrieval was blocked. When the synapse was kept at 29 degrees C for 8 min, complete vesicle depletion occurred. However, no compensatory increase in the terminal plasma membrane, either as invaginations or evaginations, was observed. Also, no internalized membranous compartment, such as cisternae or coated vesicles, appeared. No invaginations or out-pocketings were seen along the axon between release sites, and no evidence for elongation of the whole axon was found. Thus, the vesicle membrane compartment became unobservable as a result of transmitter release. Depleted synapses were observed by electron microscopy at various times after lowering the temperature, so that the process of synaptic vesicle reformation could be observed. In the first 2-3 min at 19 degrees C, gradually enlarging uncoated invaginations of the plasma membrane were observed. Between 5-10 min at 19 degrees C, these invaginations pinched off to form large cisternae. Newly formed synaptic vesicles were observed associated with these cisternae by an electron-dense material. Between 10-20 min at 19 degrees C, the number of synaptic vesicles increased, while the size of the cisternae decreased. Within 30 min, the full complement of vesicles had reappeared. No involvement of the coated vesicle pathway in synaptic vesicle reformation was observed. The data suggest that synaptic vesicle membrane is dissembled at the time of transmitter release and then is reassembled at sites along the plasma membrane and internalized in the form of large cisternae, from which new vesicles are formed. PMID- 2573699 TI - A role for cGMP during tetanus toxin blockade of acetylcholine release in the rat pheochromocytoma (PC12) cell line. AB - In order to identify the specific molecular mechanisms involved in neurosecretion, we investigated the mechanism of action of tetanus toxin, a potent presynaptic neurotoxin, in the rat adrenal pheochromocytoma PC12 cell line. It has recently been reported that tetanus toxin is a potent inhibitor of the release of depolarization-evoked 3H-acetylcholine (ACh) from nerve growth factor-differentiated PC12 cells (Sandberg et al., 1989a). In PC12 cells, as in many neural tissue preparations, cGMP accumulation in intact cells increased 6- to 17-fold when stimulated with veratridine (200 microM), carbachol (1 mM), Ba2+ (2 mM), or K+ (30 mM). Preincubation of the cells with tetanus toxin inhibits this accumulation by greater than 95%. The toxin dose-inhibition curves for 3H ACh release and cGMP accumulation are similar, with half-maximal doses of tetanus toxin seen at approximately 5 nM. The time courses for the development of the effects of tetanus on 3H-ACh release and on cGMP accumulation were also similar. Protocols which elevated intracellular cGMP levels reversed the action of the toxin. For example, evoked ACh release was restored in intoxicated PC12 cells by a 15 min exposure to 100 microM 8-bromo-cGMP. The half-maximal dose was observed at 50 microM nucleotide. Examination of the nucleotide specificity revealed that only cyclic guanine analogs were effective in reversing the effects of tetanus toxin. These results suggested that the inhibition of depolarization-evoked cGMP accumulation is causally related to the action of tetanus toxin on neurosecretion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573700 TI - Effect of structural variations of non-ionic surfactants on micellar properties and solubilization: surfactants with semi-polar hydrophobes. AB - Novel non-ionic surfactants have been synthesized in which a polar group (either an ether or a keto group) has been introduced into the hydrocarbon chain of an octadecylpolyoxyethylene glycol monoether (C18En) with an oxyethylene chain length, n, of 17-18 units. Light scattering studies have indicated aggregation numbers for these semi-polar surfactants in aqueous solution of between 55-65% of that of an unsubstituted octadecylpolyoxyethylene glycol monoether, C18E22. The solubilizing capacities of the semi-polar surfactant micelles for test compounds which were mainly solubilized at the polyoxyethylene/core interface were lower than those of C18E22 whilst solubilizates which exhibited a reasonable degree of solubility in both the interface and the micellar core showed an increased solubilization. PMID- 2573701 TI - Solubilities of adenosine antagonists determined by radioreceptor assay. AB - The practical use of many adenosine receptor antagonists is limited by poor aqueous solubility. In some cases, solubilities are so low that they are difficult to measure by conventional means. To determine solubilities of adenosine antagonists, a sensitive radioreceptor method has been developed. Solubilities in Tris buffer (pH 7.7) ranged from 141 nM for 8-(2-amino-4 chlorophenyl)-1,3-dipropylxanthine to 945 microM for the amino-substituted xanthine PD 113,297. Ratios between solubility and adenosine receptor affinity varied from 15.8 for the A2-selective antagonist HTQZ to 169,000 for PD 113,297. From literature data on functional activity, it is apparent that useful adenosine antagonist activity in-vivo is only seen in compounds with solubility/affinity ratios greater than 100. PMID- 2573703 TI - The inotropic effect of 4-aminopyridine and pH changes in rabbit papillary muscle. AB - The effects of 4-aminopyridine (4AP), and pH changes have been examined on tension responses and cyclic nucleotide levels in rabbit isolated right ventricular papillary muscles. 4AP augmented papillary muscle contractions in a concentration-dependent manner. However, this positive inotropic action was largely due to an increase in extracellular pH produced by 4AP, rather than an intrinsic activity of the drug. Increases in extracellular pH (from 5 to 9) produced graded and reproducible increases in contractile force which were not blocked by propranolol (1 x 10(-7) M) but were inhibited by verapamil in a concentration-dependent manner. The positive inotropic effects of Ca2+ were enhanced and depressed by alkaline and acidic pH, respectively. Neither 4AP nor alkaline pH significantly changed cyclic AMP concentration in rabbit papillary muscles. The cyclic GMP content, however, was increased by 4AP only and this effect was blocked by atropine. The results suggest that the positive inotropic effect associated with a rise in pH from neutrality may be due to facilitation of translocation of membrane Ca2+ and/or to increase the release of Ca2+ from sources within the cell. They also illustrate that a major component of the inotropic effect of 4AP is a result of an increase in extracellular pH. PMID- 2573702 TI - Pipequaline transport from blood to brain and liver: role of plasma protein-bound drug. AB - Brain uptake of pipequaline (45319 RP) has been studied in-vivo after a single capillary transit by intracarotid injection to rats. Pipequaline is extensively bound to plasma proteins: i.e. human serum albumin (HSA), alpha-1-acid glycoprotein (AAG), lipoproteins and blood cells, mainly erythrocytes. The dialysable drug fraction as measured in-vitro by equilibrium dialysis at 37 degrees C, was inversely related to the concentration of binding component. Similarly, the brain uptake of pipequaline was inversely related to the protein concentration of the injected solution. However, the measured brain uptake of pipequaline was higher than those predicted by in-vitro measurements of dialysable drug for all proteins and erythrocytes, except HSA. These results show that a fraction of bound pipequaline as measured in-vitro is available for transport through the blood brain barrier. HSA-bound pipequaline is an exception as it is restricted to the vascular space. Pipequaline was totally cleared by the liver through a single passage. PMID- 2573704 TI - Influence of dopaminergic and noradrenergic systems on the release of opioid peptides in guinea-pig ileum. AB - Neuroleptic drugs increase the biosynthesis and release of opioid peptides from the myenteric plexus of guinea-pig ileum. In the present work, the involvement of dopamine receptors or alpha-adrenoceptors in the release of opioids from the myenteric plexus of guinea-pig was investigated. Acute or chronic treatment with prazosin, an alpha 1-blocking drug, produced no changes in the release of these peptides. Release was also unchanged after acute or chronic treatment with the alpha 2-blocking drug yohimbine. However, treatment with domperidone, a selective dopamine receptor antagonist, resulted in an increase in the release of opioids, as did treatment with (-)-3-(3-hydroxyphenyl)-N-n-propylpiperidine ((-)-3-PPP), a dopamine autoreceptor stimulant. It is concluded that the effect of neuroleptics on the release of opioids from myenteric plexus is due to the blockade of dopamine receptors, and that interruption of dopaminergic transmission produces an increase in opioid release at this level. PMID- 2573705 TI - Effects of St-587 on the alpha-adrenoceptors in the bisected rat vas deferens. AB - The effects of the alpha-adrenoceptor agonist St-587 have been studied on the twitch responses induced by field stimulation in the prostatic portion of rat vas deferens. Moreover the drug's influence on the unstimulated prostatic and epididymal halves of rat vas deferens has also been determined. Alone and after addition of yohimbine (0.3 microM) it enhanced in a concentration-dependent manner the twitch responses in the prostatic half. Prazosin competitively antagonized (pA2 = 8.41 +/- 0.03) this effect. The enhancing effect of St-587 was not reduced in reserpinized animals. These results suggest that post-synaptic alpha 1-adrenoceptors are involved in the potentiation of twitch responses induced by St-587. When alpha 1-adrenoceptors were blocked by prazosin (0.1 microM), St-587 partially inhibited the twitch responses of the prostatic portion of rat vas deferens (Emax = 49.5 +/- 3.5%). Yohimbine completely reversed the inhibitory effects of both St-587 and clonidine. Furthermore St-587 antagonized the inhibitory effects of clonidine on twitch responses. Thus it appears that St 587 also behaves as a partial agonist of presynaptic alpha 2-adrenoceptors in this portion of rat vas deferens, but it did not induce contractions in the unstimulated prostatic half of the vas deferens. However, it competitively antagonized the alpha 1-adrenoceptor agonist phenylephrine by acting as an antagonist of prostatic postsynaptic alpha 1 adrenoceptors. These alpha 1 adrenoceptors are probably different from those that mediate the twitch enhancing response to St-587 in that portion. On the other hand, St-587 was a partial agonist of alpha 1-adrenoceptors in the epididymal half.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573706 TI - Time course of verapamil interaction with morphine effects on physiological parameters in rats. AB - The effects of subcutaneous doses of morphine and verapamil on respiratory and cardiovascular parameters have been assessed in conscious rats. Verapamil (10 mg kg-1) was injected simultaneously with morphine (16 mg kg-1) or at 10, 30, or 60 min before morphine administration. Morphine induced respiratory depression, as indicated by marked hypercapnia, hypoxia and acidosis, and caused marked tachycardia. Although morphine produced only a minor and inconsistent (but statistically significant, P less than 0.01) reduction of mean arterial blood pressure, morphine potentiated verapamil-induced hypotension. Verapamil suppressed morphine-induced hypercapnia only when injected simultaneously with morphine. Verapamil alone did not affect arterial blood gases or pH, but decreased heart rate and mean arterial blood pressure. Verapamil attenuated and delayed the maximum positive chronotropic effects of morphine at all times tested. Antagonism by verapamil of respiratory depression and tachycardia produced by morphine was unrelated to morphine levels in plasma. Thus, the explanation of verapamil-morphine interactions on respiration and cardiovascular function is not pharmacokinetic. PMID- 2573707 TI - Pharmacokinetics of cyanamide in dog and rat. AB - A pharmacokinetic study of cyanamide, an inhibitor of aldehyde dehydrogenase (E.C. 1.2.1.3) has been made in the beagle dog and Sprague-Dawley rat. Cyanamide plasma levels were determined by a sensitive high performance liquid chromatographic assay, specific for cyanamide. In the dog, i.v. administration of cyanamide at 1, 2 and 4 mg kg-1, produced a dose-dependent pharmacokinetic behaviour. Statistically significant changes were observed in plasma clearance values (12.6 to 19.7 mL kg-1 min-1), half life values (39 to 61 min) and mean residence times (50 to 79 min). Peak plasma concentrations, after oral administration of 4 mg kg-1 were achieved at 30 min and oral bioavailability was about 65%. In the rat after i.v. or oral administration, cyanamide (2 mg kg-1) had a half life of 30 min, a total plasma clearance of 117 mL kg-1 min-1 and a mean residence time of 26 min. Oral bioavailability was about 69%. PMID- 2573708 TI - Comparison of transport characteristics of amino beta-lactam antibiotics and dipeptides across rat intestinal brush border membrane. AB - The transport characteristics of amino beta-lactam antibiotics, ampicillin and cephradine, have been examined and compared with that of glycylglycine using brush border membrane vesicles isolated from rat small intestine. The initial rate of glycylglycine uptake was markedly stimulated in the presence of an inward H+ gradient compared with the uptake rates in the absence of an H+ gradient. With the same H+ gradient the stimulation of cephradine uptake was lower and ampicillin uptake was not altered. Cephradine uptake, however, was greater than that of glycylglycine in both vesicular conditions ((pH)i greater than (pH)o and (pH)i = (pH)o). Inhibitory effects of dipeptides, ampicillin and cephradine on the initial uptake of glycylglycine were also examined. Glycylglycine uptake was significantly decreased in the presence of L-phenylalanylglycine or carnosine. Ampicillin and cephradine did not alter the uptake of glycylglycine. These results suggest that the contribution of the inward H+ gradient to the permeation of ampicillin, cephradine and glycylglycine across the rat small intestinal brush border membranes is different for each of the substances examined. PMID- 2573709 TI - Carbamoylated enzyme reversal as a means of predicting pyridostigmine protection against soman. AB - The inhibition of human and mammalian red blood cell (RBC) cholinesterase (AChE) in whole blood in the presence of added pyridostigmine has been examined. After the addition of pyridostigmine to animal and human blood, red cells were separated from plasma at varying intervals and their enzyme activity measured. An apparent rate constant (ke) was derived for the reaction sequence in which carbamate is released from AChE inhibited by pyridostigmine. The constant is a complex of the rates of decarbamoylation and reinhibition of AChE in the blood sample. Rate constants were also determined for the spontaneous reactivation (ks) of carbamoylated AChE in the species studied. Values of Ks were greater than Ke in corresponding species but varied little between species. Pretreatment of animals with pyridostigmine is known to be an effective therapy against organophosphorus compounds, including soman. The ranking of ke values in mammalian blood was the same as that for the protection against soman in animals: monkey greater than guinea-pig greater than rabbit greater than rat (ke = 0.15, 0.07, 0.05, 0.02 h-1, respectively). Since ke for human blood (0.20 h-1) was greater than that of monkey, pyridostigmine pretreatment would be expected to be an effective prophylaxis for soman in humans. PMID- 2573710 TI - Ergometrine and 5-hydroxytryptamine binding sites in rat brain and myometrium. AB - Functional studies suggest that ergometrine is a partial agonist involving 5 hydroxytryptamine (5-HT) receptors in rat uterus. Ergometrine displaced [3H]5-HT from specific binding sites in rat brain, but did not displace [3H]5-HT at functionally important concentrations in rat myometrium. These binding studies indicate that the agonist and antagonist actions of ergometrine in rat uterus arise from its initial interaction with binding sites other than those for 5-HT. PMID- 2573711 TI - The effect of repeated treatment with antidepressant drugs on the thyrotropin releasing hormone (TRH)-induced hyperthermia in mice. AB - The effect of acute (single dose) or repeated (twice daily, for 14 days) administration of 10 mg kg-1 p.o. of imipramine, amitriptyline, citalopram or mianserin has been examined on the hyperthermia induced by thyrotropin-releasing hormone (TRH) (40 mg kg-1 i.p., 2, or 2 and 72 h after single or last dose of antidepressants, respectively) in mice. Both imipramine and amitriptyline, given repeatedly, potentiated the TRH response, though the effect was observed 2 but not 72 h after the last dose of those drugs. Potentiation was also found after the single dose of imipramine or amitriptyline. On the other hand, citalopram and mianserin, administered either acutely or repeatedly, did not affect the TRH induced hyperthermia. PMID- 2573712 TI - Bromocriptine-induced decrease in blood pressure in conscious spontaneously hypertensive rats: evidence for a peripheral site of action. AB - The aim of the study was to discover whether the dopamine agonist bromocriptine has a central or peripheral site of action on blood pressure. An intraperitoneal injection of bromocriptine (0.5 mg kg-1) induced a long-lasting decrease in blood pressure in conscious spontaneously hypertensive rats (SHR). This effect was blocked by peripheral pretreatment with haloperidol or domperidone, but not by central treatment with haloperidol. A central injection of bromocriptine had only minor effects on blood pressure. These results suggest that primarily peripheral, rather than central, mechanisms are involved in the hypotensive effects of bromocriptine. PMID- 2573713 TI - Absence of [3H]SCH 23390 binding sites in the rat adrenal gland. AB - The binding of D2-dopamine receptor ligand [3H]spiperone and selective D1-ligand [3H]SCH 23390 to the rat adrenal gland and striatum has been compared. [3H]Spiperone showed specific binding in both tissues revealing a Bmax of 887 fmol mg-1 protein and KD of 0.38 nM, and B of 34 fmol mg-1 protein and KD of 0.66 nM in the striatum and adrenal gland, respectively. On the other hand, [3H]SCH 23390 showed a specific binding to the striatal tissue with Bmax of 747 fmol mg-1 protein and KD of 0.70 nM, while in the adrenal tissue no specific binding was observed. These results apparently indicated only D2-dopamine receptor binding sites being present in the rat adrenal gland. PMID- 2573714 TI - Beta-adrenoceptor antagonist activities and binding affinities of timolol enantiomers in rat atria. AB - S-Timolol is an effective anti-glaucoma drug, but has potentially hazardous side effects. Recently, R-timolol, also, has been reported to be effective in lowering elevated intraocular pressure. In the present study, the beta-adrenoceptor antagonist activities and binding of R- and S-enantiomers of timolol have been examined on rat atrial preparations. The beta-antagonistic activities were investigated using spontaneously beating rat heart atria. Both timolol enantiomers inhibited (-)-isoprenaline-induced chronotropic action competitively. S-Timolol was about 54 times more potent than R-timolol. The apparent binding affinities of timolol enantiomers to beta 1- and beta 2-adrenoceptors were determined by a radioligand binding assay using (-)-[125I]iodocyanopindolol (ICYP) as a marker and CGP 20712 A as a beta 1- and ICI 118,551 as a beta 2 adrenoceptor antagonist. Both enantiomers of timolol inhibited ICYP binding in nanomolar concentrations with Hill coefficients near unity. Neither enantiomer showed selectivity between beta 1- and beta 2-adrenoceptors, but R-timolol was approximately 30 times less active than S-timolol. It is concluded that R-timolol is a relatively potent non-selective beta-adrenoceptor blocking agent, but may possibly exert a more localized beta-adrenoceptor action in the eye than S timolol, thus improving the safety of ocular timolol therapy. PMID- 2573715 TI - Putrescine reverses aconitine-induced arrhythmia in rats. AB - Putrescine, (150-300 mg kg-1 i.v.) injected into anaesthetized rats reversed aconitine-induced arrhythmia and restored sinus rhythm. In the same experimental model, quinidine and lignocaine had a transient therapeutic effect, procainamide was practically ineffective and verapamil worsened the aconitine arrhythmia, causing the death of all treated animals. These data demonstrate that putrescine has an antiarrhythmic effect in an experimental model particularly resistant to usual antiarrhythmic treatments. PMID- 2573716 TI - Disposition of alfentanil in patients receiving a renal transplant. AB - The disposition of alfentanil has been investigated in 10 anaesthetized patients with chronic renal failure undergoing kidney transplantation and compared with eight age matched anaesthetized patients with normal renal function. Plasma samples were collected to 660 min following intravenous administration of alfentanil 3-5 mg (50 micrograms kg-1). Drug concentrations were measured by RIA; and alfentanil binding to plasma proteins by equilibrium dialysis against 0.1 M phosphate buffer, pH 7.4. Alfentanil binding to plasma proteins was 87.6% (s.d. 2.0) in the patients with chronic renal failure, and 89.7% (1.2) in patients with normal renal function (P = 0.025). There was no correlation between alfentanil binding and plasma albumin, total plasma proteins, plasma urea or plasma creatinine concentrations. In both groups, the drug concentration-time profile decayed in a curvilinear manner; in the chronic renal failure patients, restoration of function did not influence the decay profile. Elimination half life, mean residence time and apparent volume of distribution at steady state were not different in the two groups of patients (mean values: 142.4 and 120.2 min; 128.5 and 136.0 min; and 40.5 and 27.6 L, respectively in chronic renal failure patients and patients with normal renal function). Total drug clearance and Vd area were significantly increased in the chronic renal failure patients: 341.9 vs 211.8 mL min-1; and 69.3 and 35.5 L. There were no differences in intrinsic clearance or apparent volume of distribution at steady state for unbound drug between the two patient groups. PMID- 2573717 TI - Bioavailability of isradipine in young and old rats: effect of mode of administration. AB - The bioavailability of isradipine has been examined in 7- and 52-week-old rats after oral (12.5 mg kg-1) or intravenous (2.5 mg kg-1) doses as a solution and administration of various doses (1.8-85.5 mg kg-1) in the diet. Serial plasma samples were obtained from each rat and the drug concentration was determined by radioimmunoassay. Absorption from the dose given by gavage was rapid but when administered in a drug diet mixture, isradipine appeared in the plasma slowly and in a manner reflecting the feeding pattern. Its absolute bioavailability from the drug-diet mixture averaged 3% over the dose range tested. By gavage its bioavailability was enhanced to 5% of dose with peak plasma values approximately 7 times higher than from a comparable dose in the diet. The low oral bioavailability of isradipine in the rat was most likely due to extensive first pass metabolism. The decline in plasma concentrations was biexponential, with a mean terminal half-life of 3.6-3.7 h after oral or intravenous dosing. The pharmacokinetic characteristics of isradipine examined were independent of the age of the rat, except that its volume of distribution decreased with age. The older rats also showed a greater inter-animal variability in isradipine bioavailability from the drug-diet mixture. PMID- 2573719 TI - Oxidation of tertiary amines by monoamine oxidases. PMID- 2573718 TI - Purification of pluronic F-68 for perfluorochemical emulsification. AB - A novel technique for purification of Pluronic F-68 has been developed involving passage through a silica-Amberlite resin column. Impurities present in commercial grade Pluronic could be readily removed, as confirmed by analysis of absorption and fluorescence emission bands. Intravenous injection of a 4% (w/v) unpurified Pluronic solution in male rats increased mean liver weight by 14% (P less than 0.05) after 24 h and increased spleen weight in female rats by 29% (P less than 0.01) after 7 days. No corresponding tissue weight changes occurred following injection of purified Pluronic solution. Haematocrit and red cell counts were also unchanged throughout. We propose that the use of purified Pluronic for perfluorochemical emulsification may reduce previously reported side-effects. PMID- 2573720 TI - HIV heterosexual transmission in hemophilia couples: lack of relation to T4 number, clinical diagnosis, or duration of HIV exposure. AB - Six (13%) of 45 female partners of 45 HIV antibody positive [Ab(+)] hemophiliacs in Western Pennsylvania have become HIV Ab(+), a mean of 10.4 months from the time of first exposure. By comparison, the mean time from first exposure to the last/recent exposure in the HIV Ab(-) female partners was 54.2 months. All six hemophiliacs who transmitted HIV to their female partners were asymptomatic, with mean T4 of 403 +/- 51/mm3 at the time of estimated seroconversion in the female; currently, their mean T4 is 203 +/- 75/mm3, which is not different from that in hemophiliac men whose partner remains HIV Ab(-) (170 +/- 22/mm3). Thus, in this population, heterosexual transmission occurs early and is unrelated to clinical or immunologic status in the hemophiliac. The risk of HIV transmission in persistently HIV Ab(-) female partners is unknown but appears low. PMID- 2573721 TI - [Cross-reactivities of active metabolites in the radioreceptor assay of an antiallergic agent, 1-(2-ethoxyethyl)-2-(hexahydro-4-methyl-1H-1,4-diazepin-1-yl) 1H-benzimidazole difumarate (KG-2413)]. AB - A new antiallergic agent, 1-(2-ethoxyethyl)-2-(hexahydro-4-methyl-1H-1,4-diazepin 1-yl)-1H- benzimidazole difumarate (KG-2413), was orally administered to guinea pigs at a dose of 2 mg/kg. The drug levels in the plasma measured by the radioreceptor assay (RRA) method were significantly higher by 7% than those by the gas chromatographic (GC) method which was selective for the intact drug. Among the metabolites of KG-2413, 1-(2-ethoxyethyl)-2-(hexahydro-1H-1,4-diazepin 1-yl)- 1H-benzimidazole (desmethyl metabolite) and 1-(2-ethoxyethyl)-5-hydroxy-2 (hexahydro-4-methyl-1H-1,4-diazepin- 1-yl)-1H- benzimidazole (5-hydroxy metabolite) had relatively high cross-reactivities of 29% and 21%, respectively, on the RRA method. However, the 5-hydroxy metabolite was not extracted with benzene under strongly basic conditions on the extraction procedure carried out prior to the receptor assay. Therefore, it was suggested that the desmethyl metabolite might affect the RRA method. In order to determine the amount of the desmethyl metabolite in the plasma, a high-performance liquid chromatographic method was established to determine the dansyl derivative of the desmethyl metabolite. By using this method, it was found that the desmethyl metabolite corresponding to 21-32% of the intact drug was present in the plasma after oral administration of KG-2413 at a dose of 2 mg/kg. Taking account of the cross reactivity (29%) and extraction ratio (83%) of the desmethyl metabolite, this active metabolite affects the RRA method to give 5.1-7.7% overestimation of the intact drug concentration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573722 TI - Infectious complications and therapy of bite wounds. AB - Bite wounds of the lower extremities present a challenge in diagnosis and management. Primary care of the initial injury remains controversial. Innocuous as these wounds may appear, severe infections are frequent. The microbial flora of the animal mouth harbor a multitude of unusual bacteria, aerobic and anaerobic, that make antibiotic selection difficult. A protocol for initial management and antibiotic selection is presented. PMID- 2573725 TI - Soldering procedure for repairing a full cast gold restoration. AB - Repairing a small hole in a cast gold restoration can often be frustrating. The method described here using platinum fold to obliterate the hole to give the solder a medium to flow across is superior to most other methods of repairing crowns and is almost foolproof. This method requires the use of the die on which the crown wa made and the use of soldering investment. PMID- 2573724 TI - Reorganization of sympathetic preganglionic connections in cat bladder ganglia following parasympathetic denervation. AB - 1. Experiments were undertaken to examine the mechanisms involved in the reorganization of sympathetic efferent pathways to the urinary bladder of the cat following chronic unilateral, parasympathetic preganglionic denervation of the bladder. 2. Electrical stimulation (10-30 Hz) of the hypogastric nerve in cats with an intact bladder innervation or on the normally innervated side of the bladder in unilaterally denervated preparations elicited low-amplitude (10-25 cmH2O) transient (10-30 s) bladder contractions and non-synaptic axonal volleys on bladder postganglionic nerves. However, after chronic (3-22 months) sacral preganglionic denervation, hypogastric nerve stimulation on the side of the denervation elicited large (60-80 cmH2O) and more sustained (4-5 min) bladder contractions as well as synaptically mediated firing on bladder postganglionic nerves. 3. The vesicoexcitatory effects of hypogastric nerve stimulation on the chronically denervated side were not altered selectively by the adrenergic blocking agent, phenoxybenzamine, but were blocked by atropine and hexamethonium suggesting that the responses were mediated by muscarinic and nicotinic cholinergic synapses. These drugs did not influence the responses elicited by hypogastric nerve stimulation on the normally innervated side of the bladder. 4. Following more extensive chronic unilateral denervation (transection of the pelvic and hypogastric nerves on one side of the bladder) stimulation of the contralateral intact pelvic nerve elicited postganglionic firing in vesical postganglionic nerves on the denervated side. This crossed excitatory pathway was not observed in normal animals or following sacral preganglionic denervation. 5. It is concluded that parasympathetic preganglionic denervation of the bladder ganglia leads to a reinnervation of the denervated cholinergic ganglion cells by sympathetic preganglionic pathways in the ipsilateral hypogastric nerve. This reinnervation results in the conversion of sympathetic inhibitory pathways to excitatory pathways in the denervated bladder. This change may contribute to the development of the autonomous hyperactive bladder seen under conditions of peripheral nerve or conus medullaris lesions of the spinal cord. PMID- 2573723 TI - On the secretory activity of single varicosities in the sympathetic nerves innervating the rat tail artery. AB - 1. Nerve terminal impulses (NTIs) and spontaneous or stimulus-evoked excitatory junction currents (SEJCs or EJCs), reflecting secretion of transmitter quanta from release sites in the sympathetic nerves of rat tail artery, were recorded by extracellular electrodes. 2. The release of transmitter quanta from single varicosities was analysed on a pulse-by-pulse basis. 3. Since the SEJCs were tetrodotoxin-resistant, and hence probably caused by single quanta, they were employed to analyse the quantal content of EJCs. 4. In the majority of recordings, EJCs were large compared to SEJCs from the same attachment, and preceded by prominent NTIs. This type of activity appeared to reflect simultaneous activation of several nerve fibres and numerous varicosities. 5. By focal stimulation, it was usually possible to improve the resolution by examining spots in which a large proportion of the suprathreshold stimuli failed to cause EJCs. Here, averaged NTIs preceding large EJCs were indistinguishable from averaged NTIs not followed by EJCs. Thus, failure of invasion by the nerve impulse was not a cause of the frequent secretory failure. 6. In these attachments the amplitude distribution of nerve stimulus-evoked EJCs was similar to that of the SEJCs and many individual EJCs could be matched in amplitude and time course by SEJCs. Thus, transmitter secretion from these sympathetic nerve varicosities seems to be basically monoquantal. 7. Under conditions when all EJCs were smaller than or equal to the largest SEJCs some characteristic EJC profiles appeared only a few times in response to several hundred suprathreshold stimuli at low frequency (0.5-1 Hz). Using tentatively these EJCs as 'fingerprints' of single quanta from particular release sites, the probability for activation of individual release sites ranges from 0.002 to 0.02. PMID- 2573726 TI - Acute neurohumoral response to electroconvulsive therapy during pregnancy. A case report. AB - The neurohumoral changes induced by electroconvulsive therapy (ECT) in pregnancy have not been described previously. In the nonpregnant adult, ECT causes an acute rise in prolactin, adrenocorticotropic hormone (ACTH), cortisol, norepinephrine, epinephrine and beta-endorphin. Because pregnancy alters the production and release of these hormones, consideration should be given to how ECT may further alter the neuroendocrine response, with possible implications for the success of treatment and the fetal response. A 30-year-old woman with a major affective disorder underwent a course of ECT beginning at 23 weeks' gestation. Serial hormonal assays of peripheral venous samples from -30 to +240 minutes were obtained during her first treatment. The prolactin, ACTH, norepinephrine, epinephrine, beta-endorphin, dopamine and oxytocin levels rose acutely and returned to baseline during observation. The maternal vital signs were stable. No increase in uterine activity or fetal heart rate abnormalities were observed during any treatment. A healthy infant weighing 2,900 g was delivered at term, with Apgar scores of 9 and 9 and no problems. We conclude that there are acute neurohumoral changes in specific hormones with ECT in pregnancy, but none of these changes appeared to adversely affect the fetus in our case. PMID- 2573727 TI - Lack of clinical benefit after treatment of systemic sclerosis with total lymphoid irradiation. AB - Six patients with systemic sclerosis and internal organ involvement were randomized to receive total lymphoid irradiation (TLI) or to serve as untreated controls. Despite evidence of profound immunosuppression, we were unable to detect any longlasting clinical benefit in the treated patients, with follow-ups ranging from 1-4 years after TLI. Moreover, the results suggest that this therapy may accelerate pulmonary and gastrointestinal deterioration in scleroderma. PMID- 2573728 TI - Acute myelocytic leukemia after the use of cyclophosphamide in the treatment of polyarteritis nodosa. AB - The use of cyclophosphamide has been associated with the delayed appearance of neoplasms. We report a woman who developed myelodysplastic syndrome leading to acute myelocytic leukemia, 4 years after receiving 13 g of cyclophosphamide over 6 months for severe polyarteritis nodosa. Cytogenetic abnormalities typical of damage induced by alkylating agents, including partial deletions of chromosomes 5 and 7, were present. This case represents the first reported occurrence of cyclophosphamide associated malignancy in polyarteritis nodosa. PMID- 2573729 TI - Chronic, painful unilateral leg swelling from vasculitis. PMID- 2573730 TI - Fatal agranulocytosis in sulfasalazine treated rheumatoid arthritis. PMID- 2573731 TI - Synthesis and structure-activity relationships of a series of anxioselective pyrazolopyridine ester and amide anxiolytic agents. AB - A series of 1-substituted 4-amino-1H-pyrazolo[3,4-b]pyridine-5-carboxylic acid esters and amides were synthesized and screened for anxiolytic activity in the shock-induced suppression of drinking (SSD) test. The compounds were also tested for their ability to displace [3H]flunitrazepam (FLU) from brain benzodiazepine (BZ) binding sites. Many compounds were active in these screens and, additionally, demonstrated a selectivity for the type 1 BZ (BZ1) receptor over the type 2 BZ (BZ2) receptor as indicated by Hill coefficients significantly less than unity and by analysis of [3H]FLU binding results from different brain regions. Based on the results of structure-activity studies of these compounds, a hypothesis was proposed to explain the structural features necessary for optimal interaction with brain BZ receptors. A detailed pharmacological evaluation of one of the most potent behaviorally active compounds (27) demonstrated it to be BZ1 selective; also, in comparison to diazepam, 27 showed minimal sedative and alcohol interactive properties at therapeutically effective doses. PMID- 2573732 TI - Synthesis and pharmacological evaluation of a series of 4-piperazinylpyrazolo[3,4 b]- and -[4,3-b][1,5]benzodiazepines as potential anxiolytics. AB - The synthesis and pharmacological evaluation of a series of pyrazolo[b][1,5]benzodiazepines are described. Some of the 4-piperazinyl-2,10 dihydropyrazolo[3,4-b][1,5]benzodiazepine derivatives demonstrated potent anxiolytic activity in the three-part operant anticonflict test in rats. Compounds 21 and 30 were more active than the clinically effective anxiolytic chlordiazepoxide in releasing conflict-suppressed behavior. This study shows a dissociation of the anxiolytic and antidopaminergic activities found in the thieno- and dibenzodiazepine derivatives flumezapine and clozapine, respectively. Examples of the three other dihydropyrazolo[b][1,5]benzodiazepine ring systems are described and evaluated for comparison and were found to be less active. PMID- 2573734 TI - Presence of two members of c-erbA receptor gene family (c-erbA beta and c-erbA2) in smallest region of somatic homozygosity on chromosome 3p21-p25 in human breast carcinoma. AB - The loss of heterozygosity of genes on the short arm of chromosome 3 (3p) in human breast carcinomas occurs in a region involved in other malignancies, including renal cell carcinoma, lung cancers, and von Hippel-Lindau disease. This finding suggests the presence of a gene(s) that plays a crucial role in multiple cancers. In our study of 84 informative (heterozygous) primary breast tumors, 30% showed losses of heterozygosity on chromosome 3. The shortest region of homozygosity in primary human breast tumor is located between the DNF15S2 and RAF1 loci in the 3p21-p25 region on the short arm of chromosome 3. This region includes at least two members of the c-erbA steroid/thyroid hormone receptor family (c-erbA beta and c-erbA2) that may be of special relevance to breast cancer. Furthermore, tumors with a loss of heterozygosity of genes on chromosome 3 were previously reported to have frequent allelic deletions on chromosome 11p and amplification of the c-myc proto-oncogene. These results highlight the occurrence of multiple genetic alterations in breast tumors. PMID- 2573733 TI - Regulation of Bacillus subtilis glutamine synthetase gene expression by the product of the glnR gene. AB - Transcription of the Bacillus subtilis gene coding of glutamine synthetase (glnA) is regulated by the nitrogen source. The glnA gene lies in an operon in which it is preceded by an open reading frame with the potential to encode a polypeptide of approximately 16,000 Mr. We have now shown that this open reading frame is utilized in vivo, that its product (GlnR) acts as a diffusible, negative regulator of gln transcription, and that GlnR is likely to be a DNA-binding protein. Certain mutations in glnR, including a large, in-frame deletion and a start codon mutation, led to high-level constitutivity of the operon; other mutations caused low-level constitutivity. These latter mutations, which affected the C terminus of GlnR, seemed to disrupt response to the nitrogen source without eliminating the ability of GlnR to bind to DNA. Wild-type GlnR by itself, however, did not impose nitrogen-dependent regulation; such regulation also required the product of glnA. A model is presented in which glutamine synthetase monitors the availability of nitrogen and imposes negative regulation by interaction with or modification of GlnR. PMID- 2573735 TI - Induction of hepatic cytochrome P-450 and xenobiotic metabolizing enzymes in rats gavaged with an Alberta crude oil. AB - Changes in body weight gain and in biochemical parameters of blood and liver were assessed in Sprague-Dawley rats after multiple oral administration of three test doses of an Alberta crude oil (ACO). Rats treated with ACO (1.25-5 ml/kg) did not show statistically significant (p greater than .05) differences from control, corn-oil treated (5 ml/kg) rats, in body weight gains, liver weight, and blood biochemical indicators of liver (alanine aminotransferase, gamma glutamyltransferase), kidney (blood urea nitrogen, creatinine), and erythrocyte (adenosine 5'-triphosphate, 2,3-diphosphoglyceric acid, reduced glutathione) cytotoxicity. Treatment with ACO, however, caused statistically significant (p less than .05) and dose-related increases from control in (1) microsomal protein and cytochrome P-450 content, and NADPH-cytochrome c reductase, aryl hydrocarbon hydroxylase (AHH), and 7-ethoxycoumarin-O-deethylase (7-ECOD) activities, and (2) cytosolic glutathione transferase activity of liver. The induction of hepatic cytochrome P-450 and xenobiotic-metabolizing enzymes in microsomes of ACO-treated rats was probably associated with dose-related changes in isozymic forms of cytochrome P-450, as evidenced by (1) appearance of a 448-nm spectral peak in microsomes of ACO-treated rats and (2) differences in the inhibition pattern of AHH and 7-ECOD activities in microsomes of control and ACO-treated rats upon treatment with metyrapone and 7,8-benzoflavone. PMID- 2573736 TI - Coated vesicles from developing and adult rat skeletal muscles contain multiple molecular forms of acetylcholinesterase. AB - The main purpose of this work was to determine which of the multiple isoforms of acetylcholinesterase (AChE) are associated with clathrin-coated vesicles (CVs) from developing and adult rat skeletal muscles. CV-enriched preparations were obtained by subcellular fractionation/equilibrium sedimentation and further purified by immunoadsorption to anti-clathrin IgG-coated Staphylococcus aureus cells. Analysis of individual AChE isoforms by velocity sedimentation ultracentrifugation showed that a) while both globular (G-forms) and asymmetric (A-forms) AChE were detected in all subcellular fractions evaluated, the CV enriched fraction contained a higher proportion of A-forms (mainly the A12 species); b) most of the AChE activity contained in such a CV fraction was recovered following immunoadsorption; c) alkaline extraction conditions (pH 8.5) which depolymerize clathrin were necessary to detect a large proportion of A forms in both the CV-enriched and immunoprecipitated preparations, while most of the G-forms (especially G1 + G2 AChE) were detected following extraction at pH 6.8; and d) comparison of AChE isoform profiles from neonate and adult muscle CV enriched fractions showed a greater concentration of A-forms in the former. These data suggest that considerable amounts of A12 and, to a lesser extent, G4 AChE are sequestered within muscle CVs which may be destined for the plasmalemma. Our findings also indicate that the relative proportions of AChE isoenzymes in rat muscle CVs vary according to the extent of muscle development and lend support to the contention that CVs participate in the externalization of functionally important AChE isoenzymes. PMID- 2573737 TI - The translational requirement for complete La Crosse virus mRNA synthesis is cell type dependent. AB - The translational requirement to prevent premature termination during La Crosse virus S mRNA synthesis was found to be cell-type dependent. This requirement was present in the BHK, HEL, and Vero cell lines we examined, but not in C6/36 mosquito cells. The cell-dependent translational requirement could be reproduced in vitro by using either cell extracts or purified virions of BHK and C6/36 cells. In the BHK reactions, the polymerase terminated predominantly at nucleotide 175 in the absence of concurrent translation and required translation to read through this position. In the C6/36 reactions, however, the polymerase reads through nucleotide 175 efficiently independent of translation. Reconstitution studies suggested that the translational requirement was due to a factor(s) present in BHK, but not in C6/36, cells. PMID- 2573738 TI - La Crosse virus nucleocapsid protein controls its own synthesis in mosquito cells by encapsidating its mRNA. AB - Within 24 to 48 h of La Crosse virus infection of mosquito cells, greater than 75% of the S mRNA was found to band in CsCl density gradients at the position of genome or antigenome nucleocapsids. The encapsidation of the S mRNA correlates with the repression of N protein synthesis in vivo, and the encapsidated S mRNA cannot be translated in vitro. Unlike genome and antigenome assembly, S mRNA assembly is a relatively slow process, which is not coupled to its synthesis. Within the encapsidated S mRNA population, three forms could be distinguished, those with intact primers which were or were not also assembled with N protein and those in which the primer and up to 3 template bases had been lost. We suggest that genome replication, but not transcription, is down regulated with time in mosquito cells for reasons that are unclear. The pool of unassembled N protein then increased to the point at which it began to interact with its own mRNA, as this mRNA also contains what is considered to be the assembly site, i.e., the conserved sequences at the 5' ends of all genome and antigenome chains. This lead to the assembly of the entire mRNA, except for the nontemplate primer. Some of the primers were then also assembled with N protein, whereas others were digested to produce truncated mRNAs. PMID- 2573740 TI - MAPHY and the two arms of HAPPHY. PMID- 2573739 TI - Peripheral T-lymphocyte activation by human T-cell leukemia virus type I interferes with the CD2 but not with the CD3/TCR pathway. AB - Human T-cell leukemia virus type I (HTLV-I) is etiologically associated with adult T-cell leukemia, an aggressive lymphoproliferative disorder, and with chronic neurological diseases. In vitro it can infect several types of cells but transforms only human T lymphocytes. We have previously shown that HTLV-I viral particles, even when noninfectious, were able to activate human resting T lymphocytes, suggesting that this activation step may be important in the initiation of the lymphoproliferative process. In the present study, we first demonstrate that in contrast to other mitogenic stimuli, HTLV-I has the unique property to activate human resting T cells in the absence of accessory cells. We then investigate the relationship between HTLV-I-induced T-cell activation and the classical well-known pathways of activation, namely, the CD3/TCR and CD2 pathways. Competitive blocking experiments were performed in which the effects of monoclonal antibodies (MAb) to the CD3/TCR complex or to the CD2 molecule were evaluated on the HTLV-I activation of T cells and compared with that obtained on phytohemagglutinin (PHA)-stimulated cells. It was found that anti-CD3 or -TCR MAb strongly suppress the proliferative response of T cells to PHA, but are significantly less efficient in inhibiting the activation initiated by HTLV-I. By contrast, MAb recognizing specific epitopes of the CD2 molecule inhibit the proliferative response of T cells to PHA or to HTLV-I to the same extent. The results provide evidence that HTLV-I virions interfere mainly with activation via CD2 but not via the CD3/TCR complex. Considering the earlier expression of the CD2 molecule on human T-cell precursors, these observations might be relevant to the characterization of the differentiation stage at which viral infection could interfere with the development and the maturation of T lymphocytes. PMID- 2573741 TI - Benzodiazepines of long and short elimination half-life and the risk of hip fracture. AB - To determine if the risk of hip fracture difference between persons receiving benzodiazepines with long (greater than or equal to 24 hours) or short (less than 24 hours) elimination half-lives, we conducted a nested case-control study among residents of the Canadian province of Saskatchewan who were 65 years of age and older. We identified 4501 cases occurring between 1977 and 1985 from computerized hospital records and 24,041 population controls. Current benzodiazepine use, defined as having filled a prescription in the past 30 days, was ascertained from computerized pharmacy records. The relative risk of hip fracture was 1.7 (95% confidence interval, 1.5 to 2.0) for current users of long half-life benzodiazepines, in contrast to that of 1.1 (95% confidence interval, 0.9 to 1.3) for current users of short half-life drugs. This finding was not altered by sex, age, calendar year, nursing home residence, or history of hospitalization. Medical record review for a sample of 189 cases suggested that this finding was not due to confounding by dementia, ambulatory status, functional status, or body mass. PMID- 2573743 TI - [Clinical study of large doses of vecuronium; duration of initial and additional doses]. AB - Neuromuscular blockade by large doses of vecuronium was investigated clinically and the duration of action of initial doses (0.2 mg.kg-1 and 0.3 mg.kg-1) and additional doses (0.01 mg.kg-1 and 0.02 mg.kg-1) were measured under enflurane nitrous oxide anesthesia using a neuromuscular transmission monitoring system (Accelograph). In group A (initial dose = 0.2 mg.kg-1, additional dose = 0.02 mg.kg-1), the time of spontaneous recovery to 25% of control twitch height (T25) and 50% of control (T50) were 63.1 and 82.0 minutes respectively. The T50 interval of the two adjacent added doses (given at the time point of 50% recovery from the previous dose) was 31.1 minutes. Reversal time from TOF ratio = 25% to 75% after administration of edrophonium was 6.1 minutes. In group B (initial = 0.3 mg.kg-1, add. = 0.01 mg.kg-1), T25, T50, T50 interval and reversal time were 122.6, 159.4, 39.2 and 4.6 minutes respectively. In group C (initial = 0.3 mg.kg 1, add. = 0.02 mg.kg-1), above values were 119.2, 145.8, 48.4 and 6.7 minutes respectively. In this study there was no obvious side effect associated with administration of large doses of vecuronium. These methods will be very useful for long surgical procedures. PMID- 2573742 TI - [Effect of induced hypotension on damage to renal tubular cells]. AB - Urinary excretion of renal tubular cell enzymes, N-acetyl-beta-D-glucosaminidase (NAG) and gamma-glutamyl transpeptidase (gamma-GTP) was evaluated to elucidate the renal cell damage before, during and after the induced hypotension with trimetaphan (TMP), nitroglycerin (TNG) and prostaglandin E1 (PGE1) under halothane-nitrous oxide-oxygen anesthesia in patients undergoing neurosurgery. Significant increases in excretion of NAG and gamma-GTP were observed in TNG group. Urinary excretion of these two enzymes in TMP group tended to increase, but its increases were not statistically significant. In PGE1 treated group, there was no tendency for urinary NAG excretion, while gamma-GTP excretion tended to decrease compared with that observed before the induced hypotension. Among these three drugs, TNG exerted the most significant effect on urinary excretion of both enzymes and TMP ranked next. Effect of PGE1 on urinary enzyme excretion was weakest. According to the general opinions, degree in stray of renal tubular cell enzyme into urine is thought to parallel with the injury of renal tubular cells. Therefore, the finding obtained in this study suggests that the induced hypotension with PGE1 has a less harmful effect on renal tubular cells. PMID- 2573744 TI - [Calcium antagonist as biological modulator in cancer chemotherapy]. PMID- 2573745 TI - [Clinical molecular-genetics for endocrine and metabolic diseases]. AB - Recently developed methods for the DNA-based diagnosis of endocrine and metabolic diseases, particularly genetic diseases, were reviewed. The molecular-diagnostic methods for a base substitution include the direct sequencing of a mutant gene, detection of a newly made RFLP site, the oligonucleotide method, the segregation analysis with RFLPs based on the linkage between and RFLP-allele and a disease locus, and the DNA/RNA mismatch method. Several examples of families with genetic diseases in which the diagnosis was successfully made are presented. Another gene mutation is a gene deletion. To detect such a deletion, the Southern hybridization method is useful. Several families with a disease, especially those with Duchenne muscular dystrophy, in which the diagnosis with deletion study was successful are presented. Finally, the method of prenatal diagnosis with the DNA technique was reviewed. PMID- 2573746 TI - [Analysis of P-glycoprotein in patients with acute leukemias by flow cytometry]. AB - The identification of a P-glycoprotein product of multidrug-resistant gene (mdr 1) was reported recently. To examine the expression of the P-glycoprotein in acute leukemias of various types, we have prepared leukemic blast cells from patients and measured their positivity of P-glycoprotein using monoclonal anti-P glycoprotein antibody (C219) by flow cytometry. P-glycoprotein is expressed in 8 out of 44 cases including leukemic blast cells but not lymphocytes and monocytes. In these cases showed drug resistance was shown clinically. In addition, the expression of the P-glycoprotein was not observed in the drug-sensitive cases and at the time of initial chemotherapy. Our results suggest that measurement of P glycoprotein in acute leukemias by flow cytometry may prove to be a valuable tool for the design of chemotherapy protocols. PMID- 2573747 TI - [Roles of vascular and renal thromboxanes in the antihypertensive effects of alpha 1 adrenoceptor antagonists]. AB - In order to assess the roles of vasoconstrictor thromboxane in the antihypertensive action of alpha 1 adrenoceptor antagonist, we explored the influences of OKY-046, a selective thromboxane inhibitor, on the antihypertensive effects of bunazosin in spontaneously hypertensive rats (SHR). 2-week antihypertensive treatment with bunazosin (0.5 mg/kg/day) did not produce a significant decrease of systolic blood pressure in SHR, as compared to untreated controls. The blood pressure reduction was associated with a decrease of PGI2/TXA2 in vascular eicosanoids generation (p less than 0.02) and an increase of TXA2 excretion in urine (p less than 0.05). A combination treatment with OKY 046 almost completely abolished the enhanced TXA2 generation in the vascular wall and kidney, which was strikingly associated with a potentiation of the blood pressure reduction by bunazosin treatment (176 vs 186 mmHg, p less than 0.01). Bunazosin directly stimulated TXA2 biosynthesis in vascular smooth muscle cells in culture in a dose-dependent manner. Thus, these data clearly indicate that bunazosin, a quinazoline derivative, enhances vasoconstrictor TXA2 system in the vascular wall and kidney possibly through direct actions, which would attenuate the antihypertensive effects of alpha 1 adrenoceptor antagonism by bunazosin treatment. PMID- 2573749 TI - [Effect of anaprilin electrophoresis using sinusoidal modulated current on central hemodynamics in patients after surgical treatment of ischemic heart disease]. AB - Rehabilitative treatment with the beta-blocker anaprilin was performed in 58 patients with coronary heart disease within early periods (on days 7 to 21) after aortocoronary bypass surgery. Group 1 patients were given oral anaprilin. Group 2 patients took the drug by sinusoidal modulated current electrophoresis. With the latter drug administration, its negative chronotropic effect and abolished adverse decrease in myocardial contractivity were detected in the patients from that group. Bearing in mind the results of pharmacokinetic studies, it was suggested that the therapeutic effect of small blood anaprilin concentrations was due to the electrophoresis-induced passage of only its therapeutically beneficial left isomer and L-propranolol glucuronide, a metabolite. PMID- 2573748 TI - [Therapeutic effects and influence on the urinary enzyme activity of human urinary trypsin inhibitor (urinastatin) in cases with acute renal failure- changes in the urinary activities of NAG and AAP]. AB - We studied the therapeutic effects of human urinary trypsin inhibitor (UTI) in 5 cases with acute renal failure, resulting from traumatic shock in 1 case, post operative shock in 2 cases, septic shock in 1 case, and dehydration in 1 case. We administered 300,000 u/day of UTI intravenously at the initial phase of acute renal failure for 7 days. We measured the activities of urinary N-acetyl-beta-D glucosaminidase (NAG), alanine aminopeptidase (AAP) and the activities of serum beta-glucuronidase, PMN-elastase serially. As a control, we also studied same markers in 5 cases with acute renal failure without the administration of UTI. We could obtain the following results. 1) Urinary activities of NAG and AAP were already elevated markedly at the onset phase of acute renal failure. 2) The administration of UTI caused a significant decrease of the activities of NAG and AAP in the urine as compared with those in the controls. 3) The administration of UTI caused also the significant suppression of the activities of beta glucuronidase and PMN-elastase in the serum. These results suggested that UTI has the protective effects on the tubular epithelial cell injuries in cases of acute renal failure. PMID- 2573750 TI - [Cryptorchism and malignant degeneration of the testis]. PMID- 2573752 TI - New molecular marker in AML: DNA-fingerprint differences between leukemic phase and remission in acute myeloid leukemia. AB - DNA-fingerprint (DNA-F) analysis, based on the polymorphism of the tandem repeats of minisatellite areas in human genome, has a capacity to reveal minor changes in dispersed areas of human genome. In this study we have applied DNA-F analysis to the detection of differences between leukemic phase and remission in acute myeloid leukemia (AML). In order to identify normal and leukemic cell populations we used two molecular probes: Jeffreys' minisatellite probes 33.6 + 33.15 and M13 wild-type phage probe. Comparison of varying minisatellite fragments between remission and diagnosis/relapse was performed by Southern blot hybridization in 21 patients with AML. The results demonstrate that Southern hybridization with minisatellite probes can detect differences in DNA-fingerprints between leukemic phase and remission in 44% of AML patients. Thus differences in DNA fingerprinting provides a new molecular marker, which can be useful in the detection of residual disease as well as in the study of the pathogenesis of AML. PMID- 2573751 TI - [The effect of prolonged antiorthostatic hypokinesia on the blood serum activity of enzymes involved in energy and plastic metabolism]. AB - In the course of 370-day antiorthostatic (-5 degrees) hypokinesia in which 9 test subjects participated the following changes in serum enzymes associated with energy metabolism inhibition were recorded: decreases of creatine phosphokinase (CPK) at the expense of its MM-isoform, isocitrate and glutamate dehydrogenases as well as increases of alanine and aspartate aminotransferases, gamma-glutamyl transferase, and lactate dehydrogenase (LDH) at the expense of an increased portion of its aerobic H-isoforms. During the recovery period these parameters returned to the norm. The counter-measures used (drugs combined with exercise) exerted a beneficial effect only in relation to CPK, gamma-glutamyl transferase and aspartate aminotransferase. PMID- 2573753 TI - Isolated gastric mucosa: an early approach to the study of the mechanism of action of gastric antisecretory agents. AB - The results of five different experiments carried out on isolated gastric mucosa were evaluated. These were: 1) Effects of antisecretory agents on (H+) and (K+) in a histamine-stimulated (4 x 10(-5)M) preparation. 2) Effects on (H+) in a preparation stimulated by dibutyryl cyclic adenosine monophosphate (dbcAMP) (6 x 10(-4)M). 3) Reversal by antipyrine (3 x 10(-2)M) of the antacid effect of antisecretory agents in a histamine-stimulated (4 x 10(-5)M) preparation. 4) Effects on the antacid activity of antisecretory agents of a pretreatment with 2 mercaptoethanol (2-ME) (2 x 10(-2)M) in a histamine-stimulated (4 x 10(-5] preparation. 5) Reversal by intraluminal increase of (K+) (up to 144.3 mM) of the antacid effect of antisecretory agents in a histamine-stimulated (4 x 10(-5)M) preparation. The technique and its application to a series of known antisecretory agents--cimetidine, ranitidine, timoprazole and omeprazole--and to other substances with antisecretory activity such as sodium thiocyanate, verapamil, trimipramine and imipramine, is described. In order to illustrate the activity of the aforementioned substances in the more classic tests of antisecretory activity, an in vivo test of inhibition of gastric secretion in pylorus-ligated rats and the in vitro tests of H2-receptor blocking activity (isolated guinea-pig atrium), anticholinergic activity (isolated guinea-pig ileum) and carbonic anhydrase (canine blood) were included. The results show that substances with different mechanisms of action behave differently in the five experiments in isolated gastric mucosa described, and these may thus be considered useful for the study of the mechanism of action of gastric antisecretory agents. PMID- 2573754 TI - Effects of some beta-adrenoceptor blockers on avoidance learning in rats. AB - In experiments on male Wistar rats, we studied the effects of some beta adrenoceptor blockers: nonselective-propranolol, pindolol and 3b (a new aminotetraline derivative); and cardioselective (beta 1)-acebutolol and talinolol, on avoidance learning. The nonselective beta-adrenoceptor blockers failed to deteriorate and in some cases even facilitated learning, estimated by the number of avoidance responses and their latencies, while the cardioselective blockers significantly impaired learning process. A significant learning impairing effect was shown by the beta 2-adrenoceptor agonist salbutamol. Similar was the effect of the mixed alpha- and beta-adrenoceptor blocker labetalol. The results are interpreted in connection with the functional role of beta 1- and beta 2- adrenoceptors in the CNS and with the importance of their balance for learning process. PMID- 2573755 TI - [Prescription of neuroleptics to mentally retarded patients is not directly criticized]. PMID- 2573756 TI - [Adrenergic beta blockaders and beta stimulants. Effects on the gastrointestinal system]. PMID- 2573757 TI - Cerebral palsy among children born during the Dublin randomised trial of intrapartum monitoring. AB - In a randomised trial involving 13,079 liveborn children intrapartum care by electronic fetal heart rate monitoring, with scalp blood sampling when indicated, was associated with a 55% reduction in neonatal seizures. Reassessment, when aged 4, of the 9 children in the intensively monitored group and 21 in the control group who survived after neonatal seizures showed that 3 such children in each group had cerebral palsy. A fourth child in the intensively monitored group with cerebral palsy had had transient abnormal neurological signs during the neonatal period. 8 other children in the intensively monitored group and 7 in the control group who had not had abnormal neurological signs in the neonatal period also had cerebral palsy. 16 (78%) of the total of 22 cases of cerebral palsy had not shown clinical signs suggestive of intrapartum asphyxia. Thus, compared with intermittent intrapartum monitoring, intensive monitoring has little, if any, protective effect against cerebral palsy. PMID- 2573758 TI - Association of low levels of mannan-binding protein with a common defect of opsonisation. AB - Failure to opsonise bakers' yeast (Saccharomyces cerevisiae) is a defect found in 5-7% of the general population. In this study, the presence of the defect was linked with low levels of mannan-binding protein (MBP), a calcium-dependent serum lectin. Purified MBP corrected the defect in a dose-dependent way in an in-vitro assay measuring the deposition of complement moieties on a mannan-coated surface. There was a highly significant correlation between the serum MBP level and the generation of C3b opsonins in a population of healthy blood donors. The median MBP level of ten children previously shown to have the functional opsonic defect was 4.9 micrograms/l (range 2.5-35.0 micrograms/l) compared with 143 micrograms/l (range 2.5-880 micrograms/l) for a paediatric control group. PMID- 2573759 TI - Precursor lesions of oesophageal cancer in young people in a high-risk population in China. AB - Young people (15-26 years) were selected from households in a population in China at high risk of oesophageal cancer on the basis of whether a case of oesophageal cancer had (166 participants) or had not (372 participants) occurred in a first degree relative. In an endoscopic survey 43.5% of the male subjects and 35.9% of the female subjects showed histological signs of chronic oesophagitis. The presence of these precursor lesions was significantly associated in a multivariate logistic model with consumption of burning hot beverages, a family history of oesophageal cancer (including second-degree relatives), infrequent consumption of fresh fruit, and infrequent consumption of dietary staples other than maize. PMID- 2573760 TI - Nifedipine for high altitude pulmonary oedema. AB - In a laboratory at 4559 m six subjects with high altitude pulmonary oedema (HAPO) characterised by clinical signs, severe hypoxaemia, widened alveolar-arterial oxygen gradient, pulmonary hypertension, and alveolar oedema on chest radiography were treated with nifedipine. Despite continued exercise at the same altitude this treatment, without supplementary oxygen, resulted in clinical improvement, better oxygenation, reduction of alveolar arterial oxygen gradient and pulmonary artery pressure, and progressive clearing of alveolar oedema. Nifedipine offers a potential emergency treatment for HAPO when descent or evacuation is impossible and oxygen is not available. The findings also suggest that hypoxic pulmonary hypertension is essential in the pathogenesis of HAPO. PMID- 2573761 TI - Double-blind study of three sodium intakes and long-term effects of sodium restriction in essential hypertension. AB - 20 patients with mild hypertension (average supine blood pressure without treatment, 164/101 mm Hg) reduced their salt intake to 50 mmol (3 g) per day for a month. They then entered a 3 month double-blind randomised crossover study of three levels of sodium intake: 200, 100, and 50 mmol per day. Blood pressure was significantly reduced on the middle and lowest sodium intakes. The average fall in blood pressure from the highest to the lowest sodium intake was 16/9 mm Hg. Patients continued to restrict their sodium intake for a further year. In 16 of the 20 patients blood pressure remained well controlled with salt restriction alone. Supine blood pressure at 1 year was 142/87 (SE 3/2) mm Hg with a 24 h urinary sodium excretion of 54 (7) mmol. These results show a progressive blood pressure fall as salt intake is reduced and that, in many patients with mild essential hypertension, blood pressure can be controlled without the need for drug therapy. PMID- 2573762 TI - Cerebral palsy, intrapartum care, and a shot in the foot. PMID- 2573763 TI - Erythropoietin reaches the pharmacy. PMID- 2573764 TI - Short-term memory impairment in chronic cannabis abusers. PMID- 2573765 TI - Invasive streptococci. PMID- 2573766 TI - Stress ulcer prophylaxis in critically ill patients. PMID- 2573767 TI - False alarms of breast cancer. AB - To assess the morbidity associated with false alarms of breast cancer, 2923 consecutive consultations for a breast disorder were reviewed. 391 women had breast cancer, which was found by accidental discovery in 57%, by breast self examination in 15%, by routine physical examination in 24%, and by screening mammography in 4%. 20% of women in whom cancers were found by physician screening had had a previous breast cancer. The pathological stages of tumours found by accident were little different from those found by intervention. 87% (2532/2923) of signs/symptoms of breast cancer were false alarms--ie, 86% (565/659) of those found by routine physical examination, 88% (406/462) breast self-examination, 93% (220/237) screening mammography, and 86% (1341/1565) accidental discovery. Spontaneous breast pain was responsible for 575 false alarms and nipple discharge for 126. 534 (20%) of the false alarms could have been avoided if routine physical examinations before the age of 45, breast self-examination before the age of 35, and screening mammography before the age of 60 had been discouraged. Another 30% of false alarms would have been avoided if the patient had realised that breast pain and nipple discharge are not usually symptoms of breast cancer. PMID- 2573769 TI - Drug companies' evidence to justify advertising. AB - Ten international pharmaceutical companies were asked by letter to supply their best evidence in support of marketing claims for seventeen products. Fifteen replies were received. Seven replies cited a total of 67 references: 31 contained relevant original data and only 13 were controlled trials, all of which had serious methodological flaws. There were four reports of changes in advertising claims and one company ceased marketing nikethamide in the third world. Standards of evidence used to justify advertising claims are inadequate. PMID- 2573768 TI - Source and route of microbial colonisation of parenteral nutrition catheters. AB - To assess the effectiveness of tunnelling the polyurethane venous catheter for parenteral nutrition in reducing the frequency of catheter microbial colonisation, and to investigate the routes taken by microorganisms colonising the central venous catheter, 109 patients were randomised to traditional subclavian catheterisation (58, group A) or to subcutaneous catheter tunnelling (51, group B). Samples were taken from patients and their nurse attendants to identify their indigenous flora. Cultures were also done of swabs from the catheter insertion site, blood, nutrient solution, segment of the catheter, and washings of the catheter hub. Intravascular segment colonisation was commoner in group A (18/58) than in group B patients (4/51), and bacterial migration from insertion site to intravascular segment was also commoner among group A (9/58) than among group B patients (1/51). Catheter hub contamination was responsible in 10 out of 22 cases of microbial colonisation; in 6 of these 10 the bacterium isolated was present on the skin of nurses who changed the bag. Contamination of the insertion site skin and of the CVC hub were equally responsible for the microbial colonisation of the intravenous segment of the catheter. PMID- 2573770 TI - Hypoglycaemia and human insulin. PMID- 2573771 TI - Plasma noradrenaline, human insulin, and hypoglycaemia. PMID- 2573772 TI - Postoperative complications in myotonic dystrophy. PMID- 2573773 TI - Alcohol and cancer of the sigmoid colon. PMID- 2573774 TI - Topical non-steroidal anti-inflammatory drugs. PMID- 2573775 TI - Failure of tetrachlorodecaoxygen anion complex to assist wound healing. PMID- 2573776 TI - Human papillomavirus and cervical cancer in Indian women. PMID- 2573778 TI - Brain tumour trends. PMID- 2573777 TI - Antibiotics for cholangitis. PMID- 2573779 TI - GM-CSF and ganciclovir for cytomegalovirus infection after autologous bone-marrow transplantation. PMID- 2573780 TI - Timing of booster doses of hepatitis B vaccine. PMID- 2573781 TI - Imported yellow fever in vaccinated tourist. PMID- 2573782 TI - Simple clinical signs for diagnosis of acute respiratory infections. PMID- 2573783 TI - Non-Hodgkin lymphoma and carbamazepine. PMID- 2573784 TI - Null results when diagnostic tests are imperfect. PMID- 2573785 TI - Octreotide and bleeding oesophageal varices. PMID- 2573786 TI - Relief of local stump pain by capsaicin cream. PMID- 2573787 TI - Haematological indices in Nigerians exposed to radioactive waste. PMID- 2573788 TI - Acute pulmonary oedema caused by crystalline methamphetamine. PMID- 2573790 TI - Pseudostatus epilepticus. PMID- 2573789 TI - Increased tetracycline resistance in gonococci in The Netherlands. PMID- 2573791 TI - Brain muscarinic receptors in Alzheimer's and Parkinson's diseases. PMID- 2573792 TI - Apartheid and AIDS. PMID- 2573793 TI - Cost-benefit analysis and prenatal diagnosis. PMID- 2573795 TI - Heatstroke and the armed forces. PMID- 2573794 TI - World Medical Association and South Africa. PMID- 2573796 TI - Ovulation induction and neural tube defects. PMID- 2573797 TI - Propofol and intensive care. PMID- 2573798 TI - Non-invasive isolation of constitutional DNA for genetic analysis. PMID- 2573799 TI - Codeine abuse. PMID- 2573800 TI - Desipramine and cyproheptadine for reversal of chloroquine resistance in Plasmodium falciparum. PMID- 2573801 TI - Neurosensory adverse effects after phenylbutazone and misoprostol combined treatment. PMID- 2573803 TI - Aerosolised pentamidine. PMID- 2573802 TI - Temporal steps in the visual field in neuropsychiatric disorders. PMID- 2573804 TI - Anal and cervical intraepithelial neoplasia. PMID- 2573805 TI - Diabetes mellitus, AIDS, and night sweats. PMID- 2573806 TI - Diagnosis of bacterial meningitis. PMID- 2573807 TI - Listeria monocytogenes and pate. PMID- 2573808 TI - Marital status of women who smoke. PMID- 2573810 TI - Nestle accused of false AIDS claim. PMID- 2573809 TI - Migraine after bone-marrow transplantation. PMID- 2573811 TI - The imidazoline-preferring receptor. AB - Evidence gathered over the past ten years supports the existence of subtypes of alpha 2-adrenoceptors. A receptor which resembles the alpha 2-adrenoceptor, called the imidazoline-preferring receptor (IPR), is virtually insensitive to catecholamines but binds selectively imidazolines and oxazolines such as idazoxan and rilmenidine. In contrast, the catecholamine-preferring alpha 2-adrenoceptor is preferentially activated by catecholamines including alpha methylnorepinephrine and epinephrine and is antagonized selectively by rauwolscine. In addition to different pharmacological profiles to agonists and antagonists, the IPR and alpha 2-adrenoceptors show differences in anatomical distribution and molecular properties. The evidence has been drawn primarily from in vitro physiological and radioligand binding studies, but is gradually extending into in vivo and even clinical studies. PMID- 2573814 TI - [The training of nurses, feldshers and midwives for work in primary medical health care]. PMID- 2573815 TI - Predicting new effective treatments of alcohol addiction on the basis of their properties of inhibition of noradrenergic activity and/or thromboxane or on the activation of the dopamine reward system and/or beta-endorphin. AB - On the basis of their properties of noradrenergic and/or thromboxane inhibition, or on their activation of the dopaminergic reward system and/or beta-endorphin, the following substances or treatments are predicted to be effective in treating alcohol or drug addiction: ginger; carbon dioxide; dietary sulfur; methionine; calcium; LHRH; high intensity light; interferon; negative ions; serotonin antagonists such as methysergide and cyproheptadine; guanabenz and guenfacine; antihistamines; head-out water immersion; X-irradiation; and forced unilateral left nostril breathing. PMID- 2573813 TI - Feeding pure docosahexaenoate or arachidonate decreases plasma triacylglycerol secretion in rats. AB - Essential fatty acid (EFA)-deficient rats were fed highly purified methyl esters of docosahexaenoate (22:6n-3), arachidonate (20:4n-6), alpha-linolenate (18:3n-3) or oleate (18:1n-9) (100 mg/day, tube fed for 3-10 days), and their plasma triacylglycerol (TG) secretion rates were measured. Secretion rates of TG into plasma were reduced by tube-feeding 22:6n-3, 20:4n-6, 18:3n-3, but not 18:1n-9, to EFA-deficient rats. A significant reduction occurred after feeding 22:6n-3 for only three days. Feeding 22:6n-3 or 18:3n-3 to EFA-deficient rats for three days also reduced the activities of liver lipogenic enzymes and sharply increased the proportions of 22:6n-3 and 20:5n-3 in liver phospholipid fractions. Mechanisms by which these EFA may reduce lipogenesis are discussed. PMID- 2573816 TI - [Current aspects of the treatment of acquired immunologic deficiency syndrome (AIDS). Report of the 5th International AIDS Conference]. PMID- 2573812 TI - Suppression of growth in a leukemic T cell line by n-3 and n-6 polyunsaturated fatty acids. AB - Proliferation in a leukemic T cell line (Jurkat) was suppressed in a dose dependent manner by n-6 and n-3 polyunsaturated fatty acids (PUFA) added to the culture medium. At high concentrations, PUFA have a cytotoxic effect on Jurkat cells. The inhibitory effect of the PUFA was not due to production of prostaglandins, and lipid peroxidation was only partly responsible. In addition to production of peroxides and aldehydes, lipid peroxidation also reduced the plasmalogen levels in these cells. The antioxidant alpha-tocopherol blocked lipid peroxidation and restored the plasmalogen levels to normal. alpha-Tocopherol did not totally restore cell proliferation although the MDA-like products in these cultures (supplemented with PUFA) were reduced to control level. Cultures supplemented with n-6 PUFA seemed to respond better to alpha-tocopherol than n-3 PUFA. This suggests that n-6 PUFA may exert their growth inhibitory effect predominantly via lipid peroxidation while different mechanisms might be operating for the n-3 PUFA. PMID- 2573817 TI - [Amplification and expression of oncogenes in human malignant tumors--organ or tissue specificity and prognosis]. AB - We investigated the amplification and expression of oncogenes in human cancerous and noncancerous tissues, and clarified the correlation between the biological characteristics and them. Amplification of c-erb B oncogene is detected in 12% of squamous cancer tissues such as cancer of the lung and esophagus, but not others. That of c-erb A and c-erb B-2 oncogenes is, specifically, detected in 20-25% of breast cancers. On the other hand, amplification of c-myc is detected in 10-20% of various cancers. EGF receptors is highly and specifically, overexpressed in squamous cancers, and the prognosis of the patients with amplification and overexpression of c-erb B oncogenes is poor. PMID- 2573818 TI - Strong homology between the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase of two species of Acetabularia and the occurrence of unusual codon usage. AB - Amino acid sequences of the small subunit of ribulose-1,5-bisphosphate carboxylase (SSU) of Acetabularia cliftonii and A. mediterranea were derived from five cDNA sequences of each of the two species of algae and by direct amino acid sequence determination of the isolated protein. An homology of more than 96% between the proteins indicates the close relationship between the two algae. All ten cDNAs in the reading frame display the termination codons TAA and/or TAG at various positions, which seem to code for the amino acid glutamine when compared with the amino acid sequence from the mature protein. This is reminiscent of proteins from ciliates where TAA and TAG also code for glutamine. PMID- 2573819 TI - Cloning of the excC and excD genes involved in the release of periplasmic proteins by Escherichia coli K12. AB - Strains of Escherichia coli K12 carrying a tolA, tolB, lky or exc mutation located at min 16.5 on the genetic map released periplasmic proteins into the extracellular medium. Wild-type genes defined by these mutations have been cloned from E. coli genomic bank made with plasmid pBR328. Subcloning experiments and complementation studies showed that lky and exc mutations were located either in the previously described tolA and tolB genes or in the newly characterized excC and excD genes. Using minicells, excC and excD gene products were identified as proteins with a molecular mass of 19 and 21 kDa, respectively. PMID- 2573821 TI - In organello replication and viral affinity of linear, extrachromosomal DNA of the ascomycete Ascobolus immersus. AB - Linear, extrachromosomal DNA's of the filamentous fungus Ascobolus immersus are localized within the mitochondria. These linear plasmids have no homology to the high molecular weight mtDNA (hmw mtDNA). For analysis of plasmid replication an in organello DNA synthesis system was developed, in which radionucleotides were incorporated into intact mitochondria. Plasmid DNA is labelled preferentially in this system. From replication analysis of a specific plasmid there is evidence of a virus-like protein-primed replication. Sequence analysis of this plasmid reveals that a viral DNA polymerase is encoded. Thus, these genetic elements presumably are viral remnants rather than true plasmids. PMID- 2573822 TI - Analysis of expression of hybrid yeast genes containing ARS elements. AB - In an attempt to devise a new assay for ARS-binding proteins we have inserted the HO ARS between the upstream activation site and the TATA region of the yeast CYC1 promoter. A marked reduction in promoter activity is observed. Inactivation of the HO ARS element by point mutation does not restore promoter activity to its original level, although a modest activation is seen. We have also inserted the HO ARS into the intron of the yeast actin gene; although there is no apparent deleterious effect on transcription, the activity of the ARS is abolished in this new environment. PMID- 2573823 TI - Synthetic isoprenoid photoaffinity labeling of P-glycoprotein specific to multidrug-resistant cells. AB - The synthetic isoprenoid N-solanesyl-N,N'-bis(3,4-dimethoxy benzyl)ethylenediamine (SDB) is known to reverse drug resistance in human multidrug-resistant KB cells. SDB inhibits the photolabeling of P-glycoprotein with the vinblastine analog N-(pazido-(3-(125)l)salicyl)-N'-beta aminoethylvindesine. We synthesized photoactive radioactive SDB and used it to photoaffinity label membrane vesicles from human KB cells and their multidrug resistant subline KB-C2 cells. A 150 to 170 kDa protein in membrane vesicles from KB-C2 cells was specifically labeled by the photoanalog of SDB. The labeled band was not detectable in parenteral drug-sensitive cells. The photolabeled 150 to 170 kDa protein was immunoprecipitated with a monoclonal antibody (C219) specific to P-glycoprotein. P-glycoprotein labeling was inhibited by anticancer agents, vinblastine, vincristine, actinomycin D, and daunomycin, with half-maximal inhibition at 2.0, 2.3, 18, and 23 microM, respectively. Only 33 and 18% of the labeling was inhibited by 100 microM Adriamycin and colchicine, respectively. The labeling was also inhibited by agents that reverse multidrug resistance, such as verapamil, reserpine, cepharanthine, and SDB. The existence of other molecules that specifically bind to 125l-SDB-photoanalog was suggested in both KB and KB-C2 membrane vesicles. The fact that we could identify the synthetic isoprenoid acceptor in membrane vesicles from multidrug-resistant cells confirms that P glycoprotein plays a role in the multidrug resistance phenotype and provides an explanation for the fact that SDB circumvents multidrug resistance. PMID- 2573820 TI - Identification and DNA sequence of tdcR, a positive regulatory gene of the tdc operon of Escherichia coli. AB - Efficient in vivo expression of the biodegradative threonine dehydratase (tdc) operon of Escherichia coli is dependent on a regulatory gene, tdcR. The tdcR gene is located 198 base pairs upstream of the tdc operon and is transcribed divergently from this operon. The nucleotide sequence of tdcR and two unrelated reading frames has been determined. The deduced amino acid sequence of TdcR indicates that it is a polypeptide of Mr 12,000 with 99 amino acid residues and contains a potential helix-turn-helix DNA binding motif. Deletion analysis and minicell expression of the tdcR gene suggest that TdcR may serve as a trans acting positive activator for the tdc operon. PMID- 2573824 TI - Discrete interactions between phosphatidylethanolamine-N-methylation and phosphatidylinositolbisphosphate hydrolysis in rat myocardium. AB - Both phosphatidylethanolamine(PE)-N-methylation and phosphatidyl-inositol bisphosphate (PI-bisphosphate) breakdown potentially modify the microdomains in the sarcolemmal lipid bilayer. In this study the possibility of a mutual interaction between the enzymes responsible for these phospholipid reactions is examined. In sarcolemma purified from rat heart, prior hydrolysis of PI lipids by exogenous specific phospholipase C inhibited (to 75, 59 and 78% of control for sites I, II and II, respectively) the PE-N-methyltransferase system. In cultured rat cardiomyocytes the addition of L-methionine, a precursor for the methyl donor S-adenosylmethionine, stimulated PE-N-methylation in a concentration (0.2-300 microM)-dependent manner. Methionine (50 microM) decreased the basal rate of PI bisphosphate hydrolysis (to 72% of control), but had no effect on the phenylephrine-stimulated PI-bisphosphate hydrolysis. Maximal activation of the PI bisphosphate breakdown by 30 microM phenylephrine did not affect the rate of PE-N methylation in the presence of exogenous methionine (50 microM). These findings support the existence of interactions, although discrete, between the enzymes involved in the PE-N-methylation and PI turnover. PMID- 2573825 TI - Redistribution of subcellular calcium in rat liver on administration of vanadate. AB - Mitochondria isolated from the livers of rats administered with sodium meta-, ortho-, or polyvanadate, but not vanadyl sulphate, exhibited enhanced Ca2+ stimulated respiration and uptake of calcium. These effects were shown also by mitochondria isolated from livers perfused with polyvanadate. The concentration of acid-soluble calcium decreased significantly in the mitochondrial fraction on vanadate treatment, while that in the cytosol showed a corresponding increase. Phenoxybenzamine, an antagonist to alpha-adrenergic receptors, effectively inhibited vanadate-induced Ca2+ mobilization, but surgical sympathectomy was without effect. This is the first demonstration of vanadate mimicking alpha adrenergic agonists in vivo. PMID- 2573826 TI - The yeast CBP1 gene produces two differentially regulated transcripts by alternative 3'-end formation. AB - CBP1 is a yeast nuclear gene encoding a mitochondrial protein that stabilizes the 5' end of cytochrome b (cob) pre-mRNA. Cytochrome b is the only mitochondrially synthesized component of the respiratory chain complex III. Since the nuclearly encoded subunits of this complex are regulated at the transcriptional level by catabolite repression, we hypothesized that CBP1 might be similarly regulated. To test the idea that transcriptional regulation of CBP1 could coordinate an increase in cytochrome b mRNA stability with an increase in nuclearly encoded complex III subunit production, we characterized the change in abundance of CBP1 mRNA during derepression on a nonfermentable carbon source. Poly(A)+ RNA from derepressed yeast cells was examined by Northern (RNA) analyses with cRNA probes from CBP1. Both 2.2- and 1.3-kilobase (kb) transcripts were detected. The 1.3-kb mRNA lacked approximately 900 nucleotides of the 3' end of the 2.2-kb mRNA, which encodes the carboxyl-terminal 250 amino acid residues of the CBP1 coding sequence. Northern analyses of RNA isolated from deletion-insertion mutants of CBP1 and from strains that overexpress CBP1 mRNA demonstrated that both mRNAs were transcribed from the CBP1 gene. Furthermore, we demonstrated that the levels of the two CBP1 mRNAs were reciprocally regulated by the carbon source in the growth medium. This is the first description of a yeast gene from which two transcripts that can encode proteins with distinctly different coding properties are generated by alternative 3'-end formation. PMID- 2573827 TI - Mammalian genes coordinately regulated by growth arrest signals and DNA-damaging agents. AB - More than 20 different cDNA clones encoding DNA-damage-inducible transcripts in rodent cells have recently been isolated by hybridization subtraction (A. J. Fornace, Jr., I. Alamo, Jr., and M. C. Hollander, Proc. Natl. Acad. Sci. USA 85:8800-8804, 1988). In most cells, one effect of DNA damage is the transient inhibition of DNA synthesis and cell growth. We now show that five of our clones encode transcripts that are increased by other growth cessation signals: growth arrest by serum reduction, medium depletion, contact inhibition, or a 24-h exposure to hydroxyurea. The genes coding for these transcripts have been designated gadd (growth arrest and DNA damage inducible). Two of the gadd cDNA clones were found to hybridize at high stringency to transcripts from human cells that were induced after growth cessation signals or treatment with DNA-damaging agents, which indicates that these responses have been conserved during mammalian evolution. In contrast to results with growth-arrested cells that still had the capacity to grow after removal of the growth arrest conditions, no induction occurred in HL60 cells when growth arrest was produced by terminal differentiation, indicating that only certain kinds of growth cessation signals induce these genes. All of our experiments suggest that the gadd genes are coordinately regulated: the kinetics of induction for all five transcripts were similar; in addition, overexpression of gadd genes was found in homozygous deletion c14CoS/c14CoS mice that are missing a small portion of chromosome 7, suggesting that a trans-acting factor encoded by a gene in this deleted portion is a negative effector of the gadd genes. The gadd genes may represent part of a novel regulatory pathway involved in the negative control of mammalian cell growth. PMID- 2573828 TI - Efficiency of utilization of the simian virus 40 late polyadenylation site: effects of upstream sequences. AB - The late polyadenylation signal of simian virus 40 functions with greater efficiency than the early polyadenylation signal, in turn affecting steady-state mRNA levels. Two chloramphenicol acetyltransferase (CAT) transient expression vectors, pL-EPA and pL-LPA, that differ only in their polyadenylation signals were constructed by using the early and late polyadenylation signals, respectively. In transfections of Cos, CV-1P, or HeLa cells and subsequent Northern blot analysis of CAT-specific RNA, approximately five times more steady state CAT mRNA was produced in transfections with pL-LPA than with pL-EPA. The basis for this difference was not related to the specific promoter used or to RNA stability. Overall, the difference in steady-state mRNA levels derived from the two plasmids appeared to be attributable to intrinsic properties of the two polyadenylation signals, resulting in distinctly different cleavage and polyadenylation efficiencies. Additionally, we found that the utilization of the late polyadenylation site was dramatically reduced by deletion of sequences between 48 and 29 nucleotides 5' of the AAUAAA hexanucleotide. This reduction of mRNA levels was shown not to be caused by altered stability of mutant precursor RNAs or mRNAs, suggesting that these upstream sequences constitute an element of the late polyadenylation signal and may cause, at least to some extent, the greater efficiency of utilization of the late polyadenylation site. PMID- 2573829 TI - Evolutionary conservation of homeodomain-binding sites and other sequences upstream and within the major transcription unit of the Drosophila segmentation gene engrailed. AB - The engrailed (en) gene functions throughout Drosophila development and is expressed in a succession of intricate spatial patterns as development proceeds. Normal en function relies on an extremely large cis-acting regulatory region (70 kilobases). We are using evolutionary conservation to help identify en sequences important in regulating patterned expression. Sequence comparison of 2.6 kilobases upstream of the en coding region of D. melanogaster and D. virilis (estimated divergence time, 60 million years) showed that 30% of this DNA occurs in islands of near perfect sequence conservation. One of these conserved islands contains binding sites for homeodomain-containing proteins. It has been shown genetically that homeodomain-containing proteins regulate en expression. Our data suggested that this regulation may be direct. The remaining conserved islands may contain binding sites for other regulatory proteins. PMID- 2573830 TI - Expression of a drug resistance gene in human neuroblastoma cell lines: modulation by retinoic acid-induced differentiation. AB - Expression of a multidrug resistance gene (mdr1) and its protein product, P glycoprotein (Pgp), has been correlated with the onset of multidrug resistance in vitro in human cell lines selected for resistance to chemotherapeutic agents derived from natural products. Expression of this gene has also been observed in normal tissues and human tumors, including neuroblastoma. We therefore examined total RNA prepared from human neuroblastoma cell lines before and after differentiation with retinoic acid or sodium butyrate. An increase in the level of mdr1 mRNA was observed after retinoic acid treatment of four neuroblastoma cell lines, including the SK-N-SH cell line. Western blot (immunoblot) analysis demonstrated concomitant increases in Pgp. However, studies of 3H-vinblastine uptake failed to show a concomitant Pgp-mediated decrease in cytotoxic drug accumulation. To provide evidence that Pgp was localized on the cell surface, an immunotoxin conjugate directed against Pgp was added to cells before and after treatment with retinoic acid. Incorporation of [3H]leucine was decreased by the immunotoxin in the retinoic acid-treated cells compared with the undifferentiated cells. These results demonstrate that whereas expression of the mdr1 gene can be modulated by differentiating agents, increased levels of expression are not necessarily associated with increased cytotoxic drug accumulation. PMID- 2573831 TI - Expression of a human multidrug resistance cDNA (MDR1) in the bone marrow of transgenic mice: resistance to daunomycin-induced leukopenia. AB - The human multidrug resistance gene (MDR1) encodes a drug efflux pump glycoprotein (P-glycoprotein) responsible for resistance to multiple cytotoxic drugs. A plasmid carrying a human MDR1 cDNA under the control of a chicken beta actin promoter was used to generate transgenic mice in which the transgene was mainly expressed in bone marrow and spleen. Immunofluorescence localization studies showed that P-glycoprotein was present on bone marrow cells. Furthermore, leukocyte counts of the transgenic mice treated with daunomycin did not fall, indicating that their bone marrow was resistant to the cytotoxic effect of the drug. Since bone marrow suppression is a major limitation to chemotherapy, these transgenic mice should serve as a model to determine whether higher doses of drugs can cure previously unresponsive cancers. PMID- 2573833 TI - Mock retroviral infection alters the developmental potential of murine bone marrow stem cells. AB - Retroviral vectors were used to introduce an activated ras gene into murine pluripotent hemopoietic stem cells. We attempted to reconstitute the hemopoietic system of lethally irradiated mice with isolated spleen colonies obtained in vivo after injection of infected bone marrow cells. Spleen colonies derived from infected bone marrow were inefficient in promoting long-term survival of irradiated hosts. This loss of reconstitutive capacity of spleen colonies was not due to the retroviral infection per se but to the in vitro culture of spleen colony precursors. Incubation for 24 h in the presence of fetal calf serum and interleukin-3 without virus-producing cells was sufficient to abolish completely the reconstitutive capacity of spleen colonies while maintaining both self renewal and pluripotential capacities of spleen colony precursors. These results show that the in vitro manipulation of stem cells that is included in current protocols for retroviral infection can modify the developmental potential of these cells. This finding clearly indicates that the use of retroviral vectors can introduce a bias in the analysis of hemopoiesis. PMID- 2573832 TI - Translational control of germ cell-expressed mRNA imposed by alternative splicing: opioid peptide gene expression in rat testis. AB - The three genes encoding the opioid peptide precursors (prodynorphin, proenkephalin, and proopiomelanocortin) are expressed in the rat testis. The sizes of the three opioid mRNAs in the testis differ from the sizes of the corresponding mRNAs in other rat tissues in which these genes are expressed. The smaller testicular proopiomelanocortin mRNA has previously been demonstrated to arise from alternative transcriptional initiation. In the present study, we found that the smaller testicular prodynorphin mRNA, expressed in Sertoli cells, results from alternative mRNA processing. Exon 2, which makes up 5' untranslated sequence, is removed from the mature transcript. Polysome analysis of brain and testis RNA indicates that the alteration of the prodynorphin leader sequence in the testis-specific transcript does not affect the efficiency of translation of this mRNA. The larger testicular proenkephalin transcript, expressed in developing germ cells, also results from alternative mRNA processing. Alternative acceptor site usage in the splicing of intron A results in a germ cell-specific proenkephalin transcript with a 491-nucleotide 5' untranslated leader sequence preceding the preproenkephalin-coding sequence. Polysome analysis indicates that this germ cell-specific proenkephalin mRNA is not efficiently translated. Mechanisms by which alternative mRNA splicing may serve to confer translational regulation upon the testicular proenkephalin transcript are discussed. PMID- 2573834 TI - Identification of a germ line transcript from the unrearranged kappa gene in human B cells. AB - A novel kappa immunoglobulin-hybridizing mRNA in cell lines derived from human B cells arrested at several stages of development has been identified. Hybridization studies demonstrate that this 1.5-kilobase mRNA species is the spliced product of a precursor germ line transcript initiating upstream of the unrearranged JKappa locus. PMID- 2573835 TI - Developmentally regulated use of alternative promoters creates a novel platelet derived growth factor receptor transcript in mouse teratocarcinoma and embryonic stem cells. AB - Embryonal carcinoma and embryonic stem cells expressed a novel form of platelet derived growth factor receptor mRNA which was approximately 1,100 base pairs shorter than the 5.3-kilobase (kb) transcript expressed in fibroblasts and other cell types. The 4.2-kb stem cell transcript was initiated within the genomic region immediately upstream of exon 6 of the 5.3-kb transcript and therefore lacked the first five exons, which encode much of the extracellular domain of the receptor expressed in fibroblasts. In stem cells, the short form was predominant, although both forms were present at low levels. Following differentiation in vitro, expression levels of the long form increased dramatically. These findings suggest that during early embryogenesis, a stem cell-specific promoter is used in a stage- and cell type-specific manner to express a form of the platelet-derived growth factor receptor that lacks much of the extracellular domain and may function independently of ligand. PMID- 2573836 TI - Discrete mutations introduced in the predicted nucleotide-binding sites of the mdr1 gene abolish its ability to confer multidrug resistance. AB - In cells stably transfected and overexpressing the mouse mdr1 gene, multidrug resistance is associated with an increased ATP-dependent drug efflux. Analysis of the predicted amino acid sequence of the MDR1 protein revealed the presence of two putative nucleotide-binding sites (NBS). To assess the functional importance of these NBS in the overall drug resistance phenotype conferred by mdr1, we introduced amino acid substitutions in the core consensus sequence for nucleotide binding, GXGKST. Mutants bearing the sequence GXAKST or GXGRST at either of the two NBS of mdr1 and a double mutant harboring the sequence GXGRST at both NBS were generated. The integrity of the two NBS was essential for the biological activity of mdr1, since all five mutants were unable to confer drug resistance to hamster drug-sensitive cells in transfection experiments. Conversely, a lysine-to arginine substitution outside the core consensus sequence had no effect on the activity of mdr1. Failure to reduce intracellular accumulation of [3H]vinblastine paralleled the loss of activity in cell clones expressing mutant MDR1 proteins. However, the ability to bind the photoactivatable ATP analog 8-azido ATP was retained in the five inactive MDR1 mutants. This result implies that an essential step subsequent to ATP binding is impaired in these mutants, possibly ATP hydrolysis or secondary conformational changes induced by ATP-binding or hydrolysis. Our results suggest that the two NBS function in a cooperative fashion, since mutations in a single NBS completely abrogated the biological activity of mdr1. PMID- 2573837 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin-inducible aryl hydrocarbon receptor-mediated change in CYP1A1 chromatin structure occurs independently of transcription. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin induces, in a receptor-dependent fashion, an increase in the accessibility of CYP1A1 chromatin to restriction endonucleases. The 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced change in chromatin structure occurs rapidly and does not require ongoing RNA or protein synthesis. The increased accessibility of chromatin DNA may facilitate its subsequent interaction with other transcription factors. PMID- 2573839 TI - Membrane depolarization evokes neurotransmitter release in the absence of calcium entry. AB - The discovery that Ca2+ is necessary for the release of neurotransmitter, the primary means by which nerve cells communicate, led to the calcium hypothesis of neutransmitter release, in which release is initiated after an action potential only by an increase in intracellular Ca2+ concentration near the release sites and is terminated (1-2 ms) by the rapid removal of Ca2+. Since then, the calcium voltage hypothesis has been proposed, in which the depolarization of the presynaptic terminals has two functions. First, in common with the calcium hypothesis, the Ca2+ conductance is increased, thereby permitting Ca2+ entry. Second, a conformational change is induced in a membrane molecule that renders it sensitive to Ca2+, and then binding of Ca2+ to this active form triggers release of neurotransmitter. When the membrane is repolarized, the molecule is inactivated and release is terminated, regardless of the local Ca2+ concentration at that moment. This hypothesis, in contrast to the calcium hypothesis, accounts for the insensitivity of the time course of release to experimental manipulations of intracellular Ca2+ concentration. Furthermore, it explains rapid termination of release after depolarization, even though Ca2+ concentration may still be high. Here we describe experiments that distinguish between these two hypotheses and find that our results support the calcium voltage hypothesis. PMID- 2573838 TI - Cocaine-induced coronary-artery vasoconstriction. AB - Intranasal cocaine is used frequently as a local anesthetic during many rhinolaryngologic procedures. Although its "recreational" use in high doses has been associated with chest pain and myocardial infarction, this association has not been established when cocaine is used in low doses as a topical anesthetic, and its effect on the coronary vasculature of humans is unknown. We studied the effects of intranasal cocaine (10 percent cocaine hydrochloride; 2 mg per kilogram of body weight) on the blood flow in and dimensions of the coronary arteries and on myocardial oxygen demand in 45 patients (34 men and 11 women, 36 to 67 years of age) who were undergoing cardiac catheterization for the evaluation of chest pain. Heart rate, arterial pressure, blood flow in the coronary sinus (measured by thermodilution), and the dimensions of the epicardial left coronary artery (measured by quantitative arteriography) were measured before and 15 minutes after the intranasal administration of saline (in 16 patients) or cocaine (in 29). No variables changed after the administration of saline. After cocaine was administered, the heart rate and arterial pressure rose, the coronary-sinus blood flow fell (from a mean [+/- SD] of 149 +/- 59 ml per minute to 124 +/- 53 ml per minute), and the diameter of the left coronary artery decreased by 8 to 12 percent (P less than 0.01 for all comparisons). No patient had chest pain or electrocardiographic evidence of myocardial ischemia after the administration of cocaine. Subsequently, the administration of the alpha-adrenergic blocking agent phentolamine caused all these values to return to base-line levels. There was no difference in response between the patients found to have disease of the left coronary artery (n = 28) and those without such disease (n = 17). We conclude that the intranasal administration of cocaine near the dose used for topical anesthesia causes vasoconstriction of the coronary arteries, with a decrease in the coronary blood flow, despite an increase in myocardial oxygen demand, and that these effects are mediated by alpha-adrenergic stimulation. It is reasonable to assume that these effects would be more pronounced at the much higher doses associated with the recreational use of cocaine. PMID- 2573840 TI - Demonstration by genetic suppression of interaction of GroE products with many proteins. AB - The way in which proteins attain and maintain their final form is of fundamental importance. Recent work has focused on the role of a set of ubiquitous proteins, termed chaperonins, in the assembly of phage and multisubunit proteins. The range of chaperonin action is unknown; they could interact with most cellular polypeptides or have a limited subset of protein partners. Included in the chaperonin family is the essential heat-shock regulated Escherichia coli groEL gene product. Over-expression of the groE operon in E. coli causes enhanced assembly of heterologously expressed ribulose bisphosphate carboxylase subunits and suppresses the heat-sensitive mutant phenotype of several dnaA alleles. It has been inferred that suppression of heat-sensitive mutations is confined to dnaA alleles and that this confinement could reflect an interaction between the groE operon products and a dnaA protein aggregate at the replication origin. We now report that multiple copies of the groE operon suppress mutations in genes encoding several diverse proteins. Our data indicate a general role for the groE operon products, the GroEL and GroES proteins, in the folding-assembly pathways of many proteins. PMID- 2573841 TI - Tolerance induction in double specific T-cell receptor transgenic mice varies with antigen. AB - The crucial role of the thymus in immunological tolerance has been demonstrated by establishing that T cells are positively selected to express a specificity for self major histocompatibility complex (MHC), and that those T cells bearing receptors potentially reactive to self antigen fragments, presumably presented by thymic MHC, are selected against. The precise mechanism by which tolerance is induced and the stage of T-cell development at which it occurs are not known. We have now studied T-cell tolerance in transgenic mice expressing a T-cell receptor with double specificities for lymphocytic choriomeningitis virus (LCMV)-H-2Db and for the mixed-lymphocyte stimulatory (MIsa) antigen. We report that alpha beta TCR transgenic mice tolerant to LCMV have drastically reduced numbers of CD4+CD8+ thymocytes and of peripheral T cells carrying the CD8 antigen. By contrast, tolerance to MIsa antigen in the same alpha beta TCR transgenic MIsa mice leads to deletion of only mature thymocytes and peripheral T cells and does not affect CD4+CD8+ thymocytes. Thus the same transgenic TCR-expressing T cells may be tolerized at different stages of their maturation and at different locations in the thymus depending on the antigen involved. PMID- 2573843 TI - [Athletic injuries of the face]. PMID- 2573842 TI - Dopamine D-2 receptor agonist-induced behavioural depression: critical dependence upon postsynaptic dopamine D-1 function. A behavioural and biochemical study. AB - The dopamine (DA) D-2 receptor agonists quinpirole (threshold dose, 0.01 mg/kg IP), pergolide (0.025 mg/kg), B-HT 920 (0.003 mg/kg) and (-)-3-PPP (4 mg/kg) produced dose-dependent locomotor depression (immobility) in mice as assessed by a subjective scoring system, with the immobility being characterized by a frozen posture. The animals were still but had their eyes open. The immobility was accompanied by reductions in sniffing, rearing and grooming. The depression (and the associated reduction in the various behaviours) produced by quinpirole (0.1 mg/kg), pergolide (0.1 mg/kg) and B-HT 920 (0.1 mg/kg) was substantially (but not always completely) reversed by the selective D-1 receptor agonist SKF38393 (up to 12 mg/kg) and the non-selective D-1 receptor agonist CY208243 (up to 3 mg/kg). The immobility induced by (-)-3-PPP (16 mg/kg) was also reversed by CY208243 and SKF38393, but the reversal was due to an increase in grooming behaviour in mice challenged with the D-1 receptor agonists, whether or not the animals had also received (-)-3-PPP. There was no reversal of the depression of rearing or sniffing. In contrast, CY208243 and SKF38393 also antagonized the immobility induced by B-HT 920, but the reversal was accompanied by at least partial reversals of the depression of sniffing, rearing and grooming. The reversal of quinpirole-induced immobility by SKF38393 and CY208243 was antagonized by SCH23390 (0.1 mg/kg). The selective D-2 receptor antagonist raclopride (0.025 to 0.4 mg/kg) could not reverse quinpirole-induced immobility. High doses of either raclopride (0.4 mg/kg) or SCH23390 (greater than 0.1 mg/kg) significantly increased immobility. Although raclopride itself (0.2 mg/kg) produced a substantial increase in DOPAC and homovanillic acid (HVA) levels in the striatum, it did not antagonize the autoreceptor mediated effects of quinpirole (0.1 mg/kg) in reducing the striatal dihydroxyphenylacetic acid (DOPAC) to DA ratio. However, the same dose of raclopride was partly effective in reducing the effects of lower doses of quinpirole (0.01 and 0.03 mg/kg) on the striatal DOPAC to DA ratio. Raclopride (0.2 mg/kg) also partially but significantly reduced the locomotor stimulant effects of d-amphetamine in reserpinized mice. Biochemical analyses in the striata indicated that CY208243 slightly retarded DA turnover (as assessed by the DOPAC/DA ratio). SKF38393 itself also slightly reduced DA turnover. In automated activity cages, using mice depleted of DA with reserpine and alpha methyltyrosine, all the D-2 receptor agonists tested, in combination with SKF38393, produced an increase in activity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2573844 TI - [The value of the registration of the position of the testis in child health services for the prevention of unnecessary orchiopexies]. PMID- 2573845 TI - Anomalies of the lumbosacral nerve roots. AB - The congenital anomalies of the lumbosacral nerve roots have frequently been found in the past as operative findings during surgery for protruded disc and are today diagnosed preoperatively with increasing frequency. They include the more cranial or more caudal origin of a nerve root, the conjoined structure of two roots and the anastomoses between two or more roots. This study reviews the large amount of literature on these anomalies and discusses their incidence, classification and embryological origin, the anatomical structure of the anomalous roots, their clinical presentation and radiological diagnosis. Anomalous nerve roots are often asymptomatic; radicular symptoms may appear in the presence of a protruded disc or lumbar stenosis, as a result of the compression or traction of the anomalous root. Myelography with water-soluble contrast media allows a good visualization of the root sheaths and thus a good diagnostic definition of these anomalies. In symptomatic cases, the surgical treatment consists of the removal of the disc hernia, associated with hemilaminectomy and foraminotomy, to obtain a good mobilization of the anomalous root; it usually results in the remission of the radicular symptoms. PMID- 2573846 TI - Does compliance predict ventricular reduction after shunting for normal pressure hydrocephalus. AB - Ventricular reduction after shunting for normal pressure hydrocephalus (NPH) was classified on the basis of magnitude and rate of reduction in 35 adult patients. Brain compliance and resistance to outflow of cerebrospinal fluid (RCSF) were determined before shunting. Rapid and marked ventricular reduction (n = 11) was associated with a significantly lower compliance than slow and moderate to marked (n = 16) or minimal to mild (n = 8) reduction. Otherwise ventricular size before as well as after shunting did not correlate with compliance or RCSF. It is concluded that both rate and magnitude of ventricular reduction after shunting for NPH vary widely. Reduced compliance seems to be the best predictor of rapid and marked reduction. PMID- 2573847 TI - Changes of cortical oxidative metabolism and cortical oxygen tension in hypoxia, anoxia and ischaemia. AB - The changes of oxidative metabolism in mitochondria with hypoxia, anoxia and ischaemia were studied by a compensated fluorometer/reflectometer in rabbits. The NADH redox state exponentially increased with the decrease of cortical oxygen tension (CoPO2) which was recorded simultaneously under systemic hypoxia or anoxia. This correlation was statistically significant (p less than 0.001). It is suggested that oxidative metabolism in mitochondria can be improved by rather small increases of CoPO2 in a severely ischaemic area. Focal cerebral ischaemia was induced by occlusion of the middle cerebral artery (MCA-O) through a transorbital approach. The NADH promptly increased with MCA-O and reached a maximal level (29.9% from the control level on average) at 20-110 s of MCA-O. Then, it partially improved to 21.2% from the control level after 5 min of MCA-O, which was statistically less than the maximal reduction level. After 5 min of MCA O, it was shown to be stable for up to 30 min of MCA-O. It appears that the partial recovery of the NADH redox state in the acute phase of arterial occlusion occurs because of the improvement of the collateral circulation demonstrated previously. This suggests one pathophysiological mechanism for transient ischaemic attacks. PMID- 2573848 TI - The effects of mild hyperoxia and/or hypertension on oxygen availability and oxidative metabolism in acute focal ischaemia. AB - The effects of oxygen inhalation (FiO2 = 0.4-0.5) and/or induced hypertension (delta MBP = around 20%) on the cortical oxygen tension (CoPO2) and the cortical oxidative metabolism (NADH/NAD redox state) in acute focal ischaemia were studied in 44 rabbits. CoPO2 was recorded by a polarographical method and NADH/NAD redox state was measured with a compensated fluorometer/reflectometer. The acute focal ischaemia was induced by the occlusion of the middle cerebral artery. With oxygen inhalation, CoPO2 improved 24.8 +/- 23.2% (mean +/- SD) in ischaemic areas where CoPO2 decreased to less than 40% of control. The oxygen inhalation also partially improved NADH levels in ischaemia by 1.5 +/- 1.6% in 8 rabbits, where NADH elevated 17.6 +/- 12.1% from the normal stage. CoPO2 and NADH redox level in ischaemia were also improved by induced hypertension. delta CoPO2/delta MBP were 1.29 +/- 1.53%/mmHg in the severely ischaemic area (less than 20% of control), 1.52 +/- 0.93 in the moderately ischaemic area (20-40% of control), and 1.03 +/- 0.62 in the mildly ischaemic area (greater than 40% of control), respectively. delta NADH/delta MBP were statistically greater in the ischaemic area than in the normal cortex (p less than 0.005). It is concluded that mild hyperoxia and induced hypertension both of which are easily employed not only can improve cortical oxygen tension but also partially restore the oxidative metabolism in acute focal ischaemia. PMID- 2573849 TI - Cerebral blood flow velocity and cerebrospinal fluid pressure after single bolus of propofol. AB - The effects of propofol on cerebral blood flow velocity, cerebrospinal fluid pressure, cerebral perfusion pressure and mean arterial pressure were studied during induction in 25 patients scheduled for elective craniotomy. Premedication consisted of only atropine sulphate 0.007 mg/kg (im) 45 min before induction. Measurements were made or derived at time zero and 1, 2, 3, 4 and 5 min after an induction dose of propofol (2.5 mg/kg). Patients were retrospectively stratified into two groups, according to cerebrospinal pressure basal values: (i) lower than 10 mmHg (10 pts) and (ii) higher than 10 mmHg (15 pts). Cerebral blood flow velocity, measured by transcranial Doppler, fell in all the patients, but the reduction was significant at 1, 2, 3 and 4 min only in the group with high CSF pressure, while it never reached the critical value of 10 cm/s. Cerebrospinal fluid pressure and mean arterial pressure decreased in both groups of patients and the fall reached a statistical significance at 1 and 2 min in the group with higher baseline CSF pressure, only at 1 min: a parallel decrease of CPP was recorded, but it was not significant. Thus propofol decreases CSF pressure without hazardous effects on cerebral blood velocity and on cerebral perfusion pressure and seems to be a suitable anaesthetic agent in controlling high cerebrospinal fluid pressure in neuroanaesthesia. PMID- 2573851 TI - Increased brain water content in pseudotumour cerebri measured by magnetic resonance imaging of brain water self diffusion. AB - Brain water self diffusion was investigated by magnetic resonance scanning in 7 patients fulfilling conventional diagnostic criteria for pseudotumour cerebri. Quantitative diffusion measurements were obtained using single spin echo pulse sequences with pulsed magnetic field gradients of different magnitude. In all patients the diffusion images showed an increased diffusion in various brain regions when compared with the diffusion coefficients for corresponding regions in healthy subjects. In 3 pseudotumour patients the increased self diffusion was localized to the periventricular regions, while 4 patients had increased diffusion in the whole brain. The findings indicate the presence of increased brain water content both intra- and extracellularly suggesting that patients with pseudotumour have two defects of pathogenetical significance: intracellular water accumulation and increased resistance to cerebrospinal fluid (CSF) outflow leading to an interstitial oedema. PMID- 2573850 TI - Simultaneous recording of tissue PCO2, interstitial pH and potassium activity in the rat cerebral cortex during anoxia and the subsequent recovery period. AB - Tissue PCO2 (carbon dioxide tension) (PtCO2), interstitial H+ (H+e) and potassium activities (K+e) were monitored in the cerebral cortex of rats during and after 2 3 min of anoxia. Anoxia was associated with systemic hypotension and caused H+e (extracellular hydrogen ion activity) to increase from pH approximately equal to 7.2 to pH approximately equal to 6.5, K+e to rise from approximately equal to 2.4 up to a maximum of approximately equal to 39 mmol/l, and PtCO2 to increase from approximately equal to 52 to approximately equal to 80 mmHg. Lactate increased from 2 to 5 mmol/kg tissue weight during anoxia and did not fall significantly after re-oxygenation for 10 min. A marked relationship existed between changes in PtCO2 and H+e. After re-oxygenation, K+e (extracellular potassium ion activity) and PtCO2 returned to the pre-anoxic level in a few minutes, whereas H+e took approximately equal to 30 min to recover. H+e recovered in a biphasic manner; a rapid decrease lasting approximately equal to 1 min preceded a much slower phase. We propose that the biphasic normalization of H+e after anoxia mainly reflects an initial and rapid washout of CO2 from brain tissue and a subsequent slow elimination of lactic acid occurring via metabolism and removal by the circulation. PMID- 2573852 TI - Noninvasive study of critical thresholds of intracranial pressure and cerebral perfusion pressure for cerebral circulation and brain function. AB - To ascertain the critical thresholds of intracranial pressure (ICP) and cerebral perfusion pressure (CPP) for cerebral circulation and brain function, the extra- and intracranial haemodynamics and electrical brain responses were evaluated noninvasively with Doppler ultrasonography and multimodality evoked potentials (MEP's) in 50 patients with severe head injury. Both extra- and intracranial blood flow velocities changed monotonically depending on the changes in ICP and CPP. They were decreased when ICP increased to 20-30 mmHg and when CPP decreased to 40-50 mmHg. The changes in elasticity index of the pulse wave of the common carotid artery was proportional to those of blood flow velocities. The frequency and degree of abnormalities of MEP's were proportionally increased with the rise of ICP and reduction of CPP. When ICP increased to higher than 31 mmHg, MEP's were classified as moderately or severely abnormal in more than 76% of the recordings. These results indicate that noninvasive study by use of Doppler ultrasonography and MEP's can provide valuable information on critical brain ischaemia and brain dysfunction in patients with acute intracranial hypertension. PMID- 2573853 TI - Accumulation of calcium in the brain following head trauma. AB - Previous studies have reported accumulation of calcium (Ca) in brain tissue of injured or ischaemic experimental animals. In the present study, head trauma (HT) was induced in the left hemisphere of rats which were subsequently sacrificed 15 min, 1, 2, 4, 24 or 48 h later. Their brains were analysed for oedema formation by the determination of specific gravity (SG), using linear gradient columns, and water content, by dry to wet weight ratio. Total tissue Ca content was measured by atomic absorption spectroscopy. These values, in both the injured and contralateral hemispheres were compared with values obtained from sham-operated rats. Specific gravity of the contused hemisphere was lower than that of the contralateral hemisphere or sham and its water content was higher, at all time points studied. Calcium content was significantly higher in the contused grey matter at 1 h, and in the grey and white matter of both hemispheres at 24 and 48 h after HT. Statistical analysis revealed excellent correlation (cc = 0.65, p less than 0.001) between Ca levels and water content in the grey matter, whenever Ca concentrations were elevated (1, 24 and 48 h). These findings suggest that in the late phase of the post-HT period, Ca accumulation might play a role, along with other mediators, in the development of brain oedema after HT. PMID- 2573854 TI - Regional cerebral blood flow with age: changes in rCBF in childhood. AB - Changes in cerebral blood flow (CBF) and regional cerebral blood flow (rCBF) throughout the entire age range including childhood were reported. The CBF in grey matter of children was markedly higher than that found in adults and showed a negative correlation with age. In contrast, the blood flow in white matter did not show a notable decrease with age. The regional distribution of CBF in children did not exhibit the frontal lobe dominance typical of adults. The rCBF pattern began to approach that of adults with growth and by 10 yr the rCBF pattern of children was similar to that of adults. PMID- 2573855 TI - Noxious and non-noxious responses in the medial thalamus of the rat. AB - Single-cell experiments were undertaken to localize and characterize the medial thalamic (MT) neurons which respond to noxious and non-noxious input in the rat. The observations demonstrated that: (1) 61 and 42% of MT neurons respond to noxious (Nox) and non-noxious (NN) stimulation, respectively; (2) MT neurons exhibit 4 cell types according to their pattern of response; Type A units were excited exclusively by Nox stimulation; Type B units were excited exclusively by NN stimulation; Type C units were excited by both (Nox and NN) stimulation, and Type D units exhibited decreases in firing rate following both stimulation modalities; (3) neurons of the parafascicularis nucleus exhibit more noxious responses (Type A units) than other medial thalamic areas. PMID- 2573856 TI - Regional donor nerves in the reinnervation of brachial plexus palsy due to upper spinal roots avulsion. AB - The prognosis for surgical treatment in cases of upper brachial plexus palsy due to spinal roots avulsion is somewhat better than in cases of total palsy. The main reasons are better possibilities for surgical reinnervation using regional donors i.e. the medial pectoral and the thoracodorsal nerves, and a shorter time span for nerve regeneration. Regional donor nerves, alone or in combination with upper intercostals and/or the spinal accessory nerve, were used in 13 cases for the reinnervation of the musculocutaneous and/or the axillary nerves. The value of the regional donors is analysed and compared with that of the spinal accessory and intercostal nerves. The value is documented throughout the results of surgical treatment with a follow-up period of at least 18 months after surgery. PMID- 2573857 TI - Role of the medullary vasomotor centre in the development of plateau waves. AB - Plateau waves can sometimes be found in various neurosurgical patients with increased intracranial pressure (ICP). In spite of the clinical importance of the waves, the precise mechanism producing them is still obscure. It has been reported that the waves are often accompanied by a reduction of arterial blood pressure (ABP) and suppression of respiration, suggesting a role of the brain stem in their development. In this study, we induced intracranial hypertension in dogs by occluding the neck veins, then stimulated the pressor and depressor areas of the brain stem, observing changes of ICP, ABP, cerebral blood flow (CBF), respiration and heart rate. Stimulation of the brain stem usually caused an increase in the ICP accompanied by variations of the ABP, CBF, respiration and heart rate. These variations were divided into two types: Type I and Type II. Type I which was induced by the stimulation of the pressor area of the brain stem comprised an arterial pressor response, an increase of CBF, hyperventilation and bradycardia. Type II which was caused by stimulation of the depressor area, included declines of the ABP and CBF, respiratory suppression and bradycardia. Of these, variations observed in Type II were similar in many respects to the plateau waves observed in clinical practice. We suggest that the depressor area of the medullary vasomotor centre may play an important role in eliciting the cerebral vasomotor reaction in the development of plateau waves in intracranial hypertension. PMID- 2573858 TI - Chemoanatomic compartments in the human bed nucleus of the stria terminalis. AB - Recent studies have indicated that peptidergic inputs to the bed nucleus of the stria terminalis are more developed in man than in rodents. To facilitate interspecies comparisons, the definition of the chemoanatomical subdivisions of the human bed nucleus of the stria terminalis was attempted. The immunocytochemistry of synenkephalin, [Met]enkephalin, somatostatin, and tyrosine hydroxylase was analysed on four verticofrontal levels in five control subjects. Four principal sectors were identified in the bed nucleus of the stria terminalis: (1) lateral, displaying an irregular patchy terminal innervation overlapping for the four markers studied; (2) central, characterized by a high density of somatostatin neurons, by pericellular basket-like formations for all markers, and by a shell of dense somatostatin innervation; (3) medial, characterized by a less dense aminergic and peptidergic innervation; and (4) lateroventral, where peptidergic (somatostatin and enkephalin) peridendritic plexuses were prominent. Double-labeling analyses showed that the somatostatin, enkephalin and tyrosine hydroxylase-like immunoreactive terminals rarely converged on the same soma or dendrite even in areas where they appeared closely interdigitated. The differences and similarities of these sectors with those defined in the rat are discussed; a marked development of the lateral and ventral bed nucleus of the stria terminalis is emphasized in man. Islands with dense peptidergic innervation, similar to the ventral bed nucleus of the stria terminalis, extended into the sublenticular substantia innominata (intercalated between the ventral pallidum and the basal magnocellular nucleus). This supports the existence of an extended amygdaloid complex from the amygdala to the bed nucleus of the stria terminalis in the human brain, as has been proposed in the rat. In relation to the literature, the present findings suggest the increasing importance of the central and lateral amygdaloid-bed nucleus of the stria terminals components and of their cortical connections in man while the medial amygdala-bed nucleus of the stria terminalis nuclei, which are preferentially connected to the olfactory system, appear less developed. PMID- 2573859 TI - D-aspartate potentiates the effects of both L-aspartate and L-glutamate on carp horizontal cells. AB - L-Aspartate, L-glutamate and D-aspartate, alone or in combination, were applied by superfusion or by atomization to the isolated carp retina while recording from cone horizontal cells. Each of these agents when applied alone depolarized horizontal cells and reduced the size of their light responses, an action which mimics the effect of the endogenous photoreceptor transmitter. Application of D aspartate in conjunction with either of the L-amino acids potentiated the effects the L-amino acids so that the threshold concentration was reduced by about five fold, compared to when the L-amino acids were applied alone. The potentiating effect of D-aspartate occurred with all types of cone horizontal cells--both L- and C-types. Furthermore, the potentiating effect of D-aspartate occurred not only in the dark but also in the presence of bright light background or Co2+ ions, conditions during which the release of photoreceptor transmitter is reduced or blocked. D-Aspartate also potentiated the depolarizing effects of the acidic amino acid cysteine sulfinate. The potentiating effect of D-aspartate can be attributed to its action as an amino acid uptake blocker in the outer retina. Thus, these findings, in themselves, cannot eliminate L-aspartate, L-glutamate or cysteine sulfinate as candidates for the carp cone transmitter. However, other evidence, previously reported, strongly suggests that L-glutamate and not L aspartate is the cone transmitter. PMID- 2573860 TI - N-methyl-D-aspartate receptors in the cortex and hippocampus of baboon (Papio anubis and Papio papio). AB - In vitro autoradiography was used to examine the N-methyl-D-aspartate receptor in the brain of a baboon species, Papio anubis, and compared to that of Papio papio which exhibits a photosensitive epilepsy. The epilepsy originates in the frontal cortex and is accompanied by an enhanced sensitivity to N-methyl-D-aspartate. In both Papio anubis and Papio papio, the density of N-methyl-D-aspartate receptors was greatest in the hippocampus, followed by associational areas including frontal cortex, and low in primary sensory areas such as the visual cortex. The receptors were concentrated in the outer cortical layers I-III, very low in layer IV except in primary visual cortex, and of intermediate density in layer V. The density of binding sites was approximately two-fold lower than previously observed in the rodent brain, whereas the affinity of the receptor for [3H]L glutamate was greater in the primate versus the rodent brain. Glycine potentiated the binding of [3H]L-glutamate in both cortex and hippocampus. No significant differences in the properties of N-methyl-D-aspartate receptors were observed between the two baboon species, suggesting that the photosensitivity of Papio papio is not due to alterations in the binding of L-glutamate to the N-methyl-D aspartate receptor complex. PMID- 2573861 TI - Embryonic entorhinal transplants project selectively to the deafferented entorhinal zone of adult mouse hippocampi, as demonstrated by the use of Thy-1 allelic immunohistochemistry. Effect of timing of transplantation in relation to deafferentation. AB - The mouse Thy-1.1/Thy-1.2 allelic marking system is used to show that transplanted embryonic entorhinal cortex can reinnervate adult host hippocampi. The projection is limited to the appropriate terminal zones--viz. the outer two thirds of the stratum moleculare of the dentate gyrus, and the stratum lacunosum moleculare of the hippocampus--and extends for up to about 2 mm into the denervated host terminal field. The reconstruction of the entorhinal projections to the host requires direct contact between the embryonic donor tissue and the denervated adult host terminal field, and is dependent upon removal of the ipsilateral host entorhinal area. In the absence of an overall deafferenting host entorhinal lesion the transplanted entorhinal area forms only small local projections which are confined to areas which would have been locally deafferented as a result of direct damage to the host entorhinal afferents (i.e. during their intrahippocampal course) by the hippocampal lesion caused at the time when the transplant was inserted. The correct relative timing of deafferentation and transplantation is vital for the formation of the transplant to-host projection. The host dendrites can be made receptive to entorhinal transplant projections by removal of the host entorhinal area at the time of transplantation. When deafferentation is performed first and transplantation is delayed, it is found that the deafferented host dendrites retain this receptivity even when deafferentation has been performed as much as two months before transplantation. Reversing the order of transplantation and deafferentation, however, shows that the transplants have only a transient ability to project to the deafferented host territory. Thus, transplants inserted and allowed to become established for one week before host deafferentation make very much reduced projections to the host, and from two weeks onwards are incapable of any detectable response to subsequent removal of the host entorhinal area. Coextensive with the formation of transplant-to-host entorhinodentate projections, the host entorhinal lesion also induces an intensification of the acetylcholinesterase staining of the host septodentate afferents in the denervated outer dentate stratum moleculare. The findings demonstrate the accurate reconstruction of a lost projection in adult brain by transplanting the appropriate type of embryonic tissue, but the results of altering the relative timing of deafferentation and transplantation raise currently unsolved questions about the nature of the competitive interactions between transplant and host axons.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2573862 TI - New strategies in dopaminergic therapy of Parkinson's disease: the use of a controlled-release formulation. PMID- 2573863 TI - Unilateral akathisia. PMID- 2573864 TI - Gastrointestinal pharmacology review and anesthetic application to the combat casualty. AB - Pulmonary aspiration of gastric content remains the number one cause of anesthetic mortality. The combat casualty due to the nature of wounds inflicted is at risk for pulmonary aspiration on induction and emergence from anesthesia. The use of gastrointestinal pharmacology agents can reduce this risk significantly. Histamine-2 receptor blocking agents Cimetidine or Ranitidine in combination with metaclopramide, which enhances LES tone, facilitate management of induction and emergence in the traumatized patient. PMID- 2573865 TI - Hypoxia/hypotension evoked release of glutamate and aspartate from the rat cerebral cortex. AB - Glutamate and aspartate levels in cerebral cortical superfusates rose rapidly after introduction of artificial cerebrospinal fluid into cortical cups to reach stable levels within 5-10 min. These equilibrated levels (glutamate 1.33 +/- 0.1 microM; aspartate 0.29 +/- 0.04 microM), which are very similar to those in fourth ventricular CSF (glutamate, 1.5 +/- 0.2 microM; aspartate, 0.27 +/- 0.03 microM), are likely to reflect basal interstitial amino acid levels in the superficial layers of the cortex. During and following brief (5 min) hypoxemic/hypotensive episodes, release of both excitatory amino acids into the cortical superfusates was enhanced. The finding of an hypoxia/hypotension-evoked release of excitatory amino acids is consistent with recent speculation that these agents may play an important role in the destruction of neurons in ischemic regions of the brain. PMID- 2573866 TI - Excitatory amino acid-mediated components of synaptically evoked input from dorsal roots to deep dorsal horn neurons in the rat spinal cord slice. AB - The participation of N-methyl-D-aspartate (NMDA) and non-NMDA receptors in the responses of deep dorsal horn neurons to single shock stimulation of dorsal roots was investigated using current- and voltage-clamp techniques. In the presence of Mg2+, superfusion of rat spinal slices with 6-cyano-2,3-dihydroxy-7 nitroquinoxaline (CNQX), a potent antagonist of non-NMDA receptors, reversibly blocks fast excitatory synaptic responses elicited by low-frequency stimulation of dorsal roots and to a greater extent the responses to quisqualate than to kainate or NMDA. The synaptic response elicited in a zero-Mg2+ medium is less sensitive to CNQX. The CNQX-resistant component is however abolished by D-APV, a selective antagonist of NMDA receptor. Under voltage-clamp, the excitatory postsynaptic currents also showed an initial fast (CNQX-sensitive) and a late slow (2-amino-5-phosphonovalerate (APV)-sensitive, Mg2+-sensitive) component, both of which had similar thresholds but differed in their latency, time-to-peak and duration. These results support the concept that both non-NMDA and NMDA receptor channels are present in a majority of deep dorsal horn neurons and could be simultaneously activated by transmitter released from stimulated primary afferents. PMID- 2573867 TI - Participation of excitatory amino acid receptors in the slow excitatory synaptic transmission in the rat spinal dorsal horn in vitro. AB - The participation of excitatory amino acid (EAA) receptors in the responses of deep dorsal horn neurons to repetitive stimulation of dorsal roots was investigated using a spinal slice preparation and current-clamp and voltage-clamp techniques. Using EAA receptor and substance P (SP) receptor antagonists and current-clamp, slow excitatory synaptic response evoked by 10-20 Hz stimulation consisted of two depolarizing components: an initial component lasting 1-5 s and a late-one of 1-3 min duration. The initial and late components of the slow excitatory postsynaptic currents (EPSCs) can also be distinguished on the basis of their voltage-dependence and sensitivity to Mg2+ ions, D-2-amino-5 phosphonovalerate (D-APV) and 6-cyano-2,3-dihydroxy-7-nitroquinoxaline (CNQX). In the presence of Mg2+, the initial component of the slow EPSC increased with membrane hyperpolarization, whereas the late component decreased. In a zero-Mg2+ medium, the initial component was potentiated, but the late component was reduced, or unchanged. CNQX reduced the initial component. In a zero-Mg2+ solution, or at membrane potentials positive to -55 mV in 1 mM Mg2+, D-APV reduced or even abolished the initial component, whereas the late component was not modified by D-APV. We propose that slow excitatory synaptic response evoked in deep dorsal horn neurons by repetitive stimulation of primary afferents has two components, an initial transient component that requires activation of N methyl-D-aspartate (NMDA) and non-NMDA receptors, and a late longer-lasting peptidergic component that has been already described (Brain Res., 290 (1984) 336 341. PMID- 2573868 TI - Expression of tyrosine hydroxylase mRNA in transplanted fetal dopamine neurons. AB - Expression of genes coding for synthesis of secretory products has been shown to be an important index of neuronal activity. Gene expression in transplanted fetal substantia nigra (SN) was examined for the first time, utilizing in situ hybridization with a probe for tyrosine hydroxylase mRNA. Three months after implantation, the grafts contained many labeled neurons. Compared to host SN neurons, the grafted dopaminergic cells expressed more message for the enzyme, while they showed lower amounts of the enzyme itself. This result suggests that a molecular approach applied to neural transplantation can detect important if subtle differences in graft cell activity. PMID- 2573869 TI - Lewy bodies in tyrosine hydroxylase-synthesizing neurons of the human cerebral cortex. AB - A population of neurons situated in the human cerebral neocortex contains mRNA coding for tyrosine hydroxylase, the key enzyme for catecholamine biosynthesis. Phosphorylated neurofilament-containing cytoplasmic inclusions occur in these neurons in diffuse Lewy body disease, indicating a tendency for selective involvement that is shared with subcortical catecholamine-containing neurons. These findings are relevant to the pathophysiology of several neurologic and psychiatric illnesses in which the monoamine-containing neurons of the neocortex may participate. PMID- 2573870 TI - Co-localization of tyrosine hydroxylase and GABA immunoreactivities in human cortical neurons. AB - Samples of human cerebral cortex were stained immunocytochemically for tyrosine hydroxylase (TH) and gamma-aminobutyric acid (GABA). TH-positive neurons were in small number and predominated in the deep infragranular layers V-VI contrasting with numerous GABA-positive neurons scattered in all layers. Co-localization of TH- and GABA-like immunoreactivities in a single cell was studied by the double immunolabeling technique with the elution-restaining procedure. Only 50% of the TH-positive neurons also expressed GABA-like immunoreactivities. The two markers were detectable in the somata and not in the processes of the cells. The double labeled cells were mainly fusiform and medium-sized and were observed in layer VI. These observations suggest that the TH-positive cells form a mixed neuronal population, only a part of which corresponds to the GABAergic class of intrinsic interneurons. PMID- 2573871 TI - Giving narcotics for pain. PMID- 2573872 TI - A treatment for tardive dyskinesia in a complete denture patient. A case report. PMID- 2573873 TI - [What is social pharmacology, what are its goals?]. AB - Sociopharmacology is based on interdisciplinary research involving pharmacology, epidemiology, social medicine and sociology. It aims at finding out the social determinants of drug intake. How this goal is approached stepwise is shown in the case of the benzodiazepine group of active substances. PMID- 2573874 TI - Plasma somatostatin activity in medullary cancer of the thyroid. AB - In a group consisting of 50 adults and 5 children with diagnosed or suspected medullary cancer of the thyroid, plasma somatostatin levels were measured in 77 samples with a sensitive radioimmunoassay. Only 2 patients had clearly enhanced plasma somatostatin immunoreactivity, both with highly active aggressive tumors. Other patients had plasma somatostatin levels of under 20 ng/l or undetectable ones. No evidence of response of somatostatin levels to pentagastrin stimulation was found. PMID- 2573875 TI - Alpha2-adrenergic receptors in rat and rabbit eye: a tritium-sensitive film autoradiography. AB - Using a tritium-sensitive film and [3H]-clonidine, alpha 2-adrenergic receptors were localized in entire eye sections of rabbits and rats. This radioactive agonist specifically labelled three structures in albino animals: ocular muscles, ciliary processes and retina. In pigmented animals, uvea with melanin was intensively but nonspecifically labelled. Microscopic analysis of autoradiograms obtained with iris-ciliary bodies, revealed that the major localization of binding sites was in the ciliary process epithelium. In displacement studies, [3H]-clonidine binding was inhibited by alpha 2-adrenergic compounds while alpha 1- or beta-selective drugs had no inhibitory effects. PMID- 2573876 TI - Limiting dilution analysis of the T cell response to Plasmodium chabaudi chabaudi in mice. AB - A limiting dilution assay system was developed in order to measure the in-vitro T cell response to antigens of the erythrocytic stages of Plasmodium chabaudi. The conditions of the assay are such that only CD4+ T cells are able to respond. The assay allows the determination of the frequencies of T cells which proliferate and/or which develop into helper cells for antibody production during a primary infection. A specific response from splenic T cells can be measured as early as 7 days after infection, and is still significant 3 months after injection of P. chabaudi. At all times the frequency of proliferating cells was greater than the precursor frequency of T helper cells. This suggests that a proportion of CD4+ T cells in this assay, although they respond to malarial antigen, do not develop into helper cells for antibody production. This limiting dilution assay will be a useful method by which to evaluate the functional heterogeneity of the CD4+ T cell response to malaria antigens. PMID- 2573877 TI - Two remote glucocorticoid responsive units interact cooperatively to promote glucocorticoid induction of rat tyrosine aminotransferase gene expression. AB - Tyrosine aminotransferase (TAT) gene transcription is specifically activated by glucocorticoid hormones in liver cells. This regulation involves a glucocorticoid responsive region located 2,500 bases upstream from the transcription start site of the rate gene. By transient transfection of TAT-CAT fusion genes into a rat hepatoma cell line expressing the TAT gene we found that this region promotes only 30% of the glucocorticoid stimulation. We have identified a new cis-acting region far upstream (-5,400) from the transcription start site that is essential to achieve the physiological level of glucocorticoid stimulation of endogenous TAT gene expression. This region corresponds to a tissue-specific DNAse I hypersensitive site which is constitutive despite the fact it possesses a glucocorticoid receptor binding site. It is by itself almost inactive on a promoter but it cooperatively enhances the action of the proximal glucocorticoid responsive region. Its activity requires both the glucocorticoid receptor binding site and its flanking sequences. PMID- 2573880 TI - RFLPs for ATP1BL1 (beta subunit Na+/K+ ATPase pseudogene) on chromosome 4. PMID- 2573878 TI - Duplication and transcription of procyclin genes in Trypanosoma brucei. AB - The genes encoding procyclin, the major glycoprotein expressed on the surface of procyclic forms of Trypanosoma brucei, comprise a multigene family. It has previously been demonstrated that procyclin genes in cloned trypanosome strains from Kenya and Uganda show restriction fragment polymorphisms. A detailed study of the Kenyan strain 227 has revealed that procyclin genes are arranged in tandem at 3 distinct loci (Pro A, B and C) and that the polymorphism is due to the duplication of 1.3 kb in the Pro A locus, which has generated an additional procyclin gene. Northern blot analysis has shown that at least 2 loci are transcribed and that a minimum of 3 procyclin genes are expressed within a cloned line. The transcription of procyclin genes is resistant to 1 mg ml-1 alpha amanitin, whereas that of the 5' flanking gene in the Pro A locus is sensitive. This observation suggests that the two genes form part of separate transcription units with a promoter between them. PMID- 2573879 TI - Nucleotide sequence of 10 kilobases of rat tyrosine aminotransferase gene 5' flanking region. PMID- 2573881 TI - A ScaI RFLP demonstrated for the GRO gene on chromosome 4. PMID- 2573882 TI - RFLP for the human retinoic acid receptor gene RAR-beta. PMID- 2573883 TI - A BamHI-polymorphism is detected by a genomic p53-clone (pBHP53). PMID- 2573884 TI - A BglII RFLP at the lipoprotein lipase gene. PMID- 2573885 TI - A TaqI RFLP of the human plasminogen gene. PMID- 2573886 TI - TaqI RFLP identified by probe 1A1 [DXS374] at Xq28. PMID- 2573887 TI - Lack of correlation between BglII RFLP in the human interleukin 6 gene and rheumatoid arthritis. PMID- 2573889 TI - Isolation and mapping of a polymorphic DNA sequence (CJ52.193) on chromosome 11 [D11S384]. PMID- 2573890 TI - Isolation and mapping of a polymorphic DNA sequence (CJ52.5) on chromosome 11 [D11S386]. PMID- 2573888 TI - Two BglII RFLPs of the human alpha-globin gene cluster in the American sickle cell population. PMID- 2573891 TI - Isolation and mapping of a polymorphic DNA sequence (CJ52.96) on chromosome 16 [D16S157]. PMID- 2573892 TI - Isolation and mapping of a polymorphic DNA sequence (pCJ52.102T1) on chromosome 11 [D11S387]. PMID- 2573893 TI - Comparative effects of Pro-Leu-Gly-NH2 and cyclo(Leu-Gly) administered orally on the development of tolerance to the analgesic effect of morphine in the rat. AB - Comparative effects of Pro-Leu-Gly-NH2 (MIF) and cyclo(Leu-Gly) (CLG) administered orally at different stages of chronic morphine treatment on the development of tolerance to the analgesic effect of morphine in the rat were determined. Male Sprague-Dawley rats were implanted with either 6 placebo or morphine pellets during a 7-day period. Implantation of morphine pellets resulted in the development of a high degree of tolerance as evidenced by a decrease in the analgesic response to morphine. Administration of CLG (8 and 16 mg/kg/day) on day 5, 6 and 7 of implantation inhibited the development of tolerance to morphine but 4 and 32 mg/kg doses had no effect. Further, CLG (2 mg/kg/day for 7 days) inhibited the development of tolerance but higher doses (4 and 8 mg/kg) had no effect. MIF (26 and 52 mg/kg) administered orally on the last three days of the implantation schedule inhibited the development of tolerance to morphine. MIF (6.5 mg/kg/day for 7 days) inhibited the development of tolerance but the higher doses had no effect. Concurrent administration of MIF (6.5 mg/kg) and CLG (2 mg/kg) for seven days failed to inhibit the development of tolerance. A single dose of MIF or CLG administered a day before the assessment of tolerance did not affect the morphine tolerance. Thus, even after a significant degree of tolerance to morphine had developed, neuropeptides like MIF and CLG given orally, in appropriate doses, can inhibit development of tolerance to morphine and restore the analgesic effect of morphine. PMID- 2573894 TI - Partial coexistence of growth hormone-releasing hormone and tyrosine hydroxylase in paraventricular neurons in rats. AB - Immunocytochemistry revealed growth hormone-releasing hormone (GRF)-containing cells (4-10/50 microns thick coronal sections) in the ventral portion of the medial parvicellular subdivision of the paraventricular nucleus (PVN) in the rat. In the same region we also detected about the same number of neurons containing the mRNA that encodes GRF using in situ hybridization histochemistry. Fluorescence double labelling immunohistochemistry showed the presence of GRF and tyrosine hydroxylase (TH) in the same PVN neurons. The overlap between the two populations of cells is only partial: less than half of TH-positive cells contain GRF and vice versa. PMID- 2573896 TI - Treating acute schizophrenic illnesses. AB - Schizophrenia has a lifetime expectancy of about 1 per cent and the GP can be involved in management at all stages. One important role is counselling and support for the family. In the drug treatment of schizophrenia, it is useful to become familiar with two or three drugs. PMID- 2573895 TI - Aplysia californica neurons R3-R14: primary structure of the myoactive histidine rich basic peptide and peptide I. AB - The R3-R14 neurons of the marine mollusc Aplysia are neuroendocrine cells that express a gene encoding peptides I, II and histidine-rich basic peptide (HRBP), a myoactive peptide that excites Aplysia heart and enhances gut motility in vitro. Peptide II has been chemically characterized (35), but the complete primary structures of peptide I and HRBP have not been established by amino acid sequence analysis. HRBP, peptide I, and the prohormone (proHRBP) were therefore purified from acid extracts of Aplysia californica neural tissue using sequential gel filtration and reverse-phase high-performance liquid chromatography and chemically characterized. Amino acid sequence analysis demonstrated that HRBP was a 43-residue peptide whose sequence was: less than Glu-Val-Ala-Gln-Met-His-Val Trp-Arg-Ala-Val-Asn-His-Asp-Arg-Asn-His-Gly- Thr-Gly - Ser-Gly-Arg-His-Gly-Arg Phe-Leu-Ile-Arg-Asn-Arg-Tyr-Arg-Tyr-Gly-Gly-Gly- His-Leu - Ser-Asp-Ala-COOH. Compositional and sequence analyses of peptide I and proHRBP demonstrated that peptide I was a 26-residue peptide with the following sequence: NH2-Glu-Glu-Val Phe-Asp-Asp-Thr-Asp-Val-Gly-Asp-Glu-Leu-Thr-Asn-Ala- Leu-Glu-Ser-Val-Leu-Thr-Asp Phe-Lys-Asp-COOH. These results demonstrated that the pro-HRBP sequence predicted by nucleotide sequence analysis of a cDNA clone (24) was in fact synthesized in R3-R14 neurons. Hydrophilicity and hydrophobicity profiles of preproHRBP, combined with charge distribution profiles and predictive secondary structural analysis, showed that cleavage at dibasic sequences was strongly associated with peaks of hydrophilicity in alpha-helical regions of the preprohormone. PMID- 2573897 TI - Management of chronic schizophrenia. AB - Schizophrenia is usually a chronic illness characterised by a series of relapses. Long-term management aims to achieve symptomatic remission for as long as possible. Maintenance drug therapy should be considered and is about one-third of the acute dose. Support in the community as offered by multi-disciplinary teams at day centres and community organisations plays an important part. PMID- 2573898 TI - Spiked! A cross-country injury. AB - In this new series, GPs with a special interest in a particular sport describe the practical management of a typical injury. Frank Newton begins with the spiking injury seen in cross-country running. PMID- 2573899 TI - Comparison of the effects of the novel antipsychotic agent remoxipride on dopamine and noradrenaline turnover in the rat brain. AB - The ex vivo effects of the antipsychotic dopamine receptor antagonist remoxipride on dopamine and noradrenaline turnover in the rat brain have been compared. Remoxipride (5-125 mumol/kg intraperitoneally) produced large increases in striatal DOPAC and HVA concentrations with only maginal effects on striatal, hippocampal and hypothalamic MHPG concentrations. Both remoxipride (5.6 mumol/kg intraperitoneally) and haloperidol (0.23 mumol/kg intraperitoneally) increased the rate of striatal dopamine disappearance following inhibition of tyrosine hydroxylase by H 44/68 without corresponding effect on hypothalamic and frontal cortical noradrenaline disappearance. It is concluded that at doses of remoxipride producing compensatory increases in dopamine turnover, there is little or no effect on noradrenaline turnover, confirming the in vitro dopamine:noradrenaline selectivity of this compound. PMID- 2573901 TI - Vertical disparity, egocentric distance and stereoscopic depth constancy: a new interpretation. AB - There has long been a problem concerning the presence in the visual cortex of binocularly activated cells that are selective for vertical stimulus disparities because it is generally believed that only horizontal disparities contribute to stereoscopic depth perception. The accepted view is that stereoscopic depth estimates are only relative to the fixation point and that independent information from an extraretinal source is needed to scale for absolute or egocentric distance. Recently, however, theoretical computations have shown that egocentric distance can be estimated directly from vertical disparities without recourse to extraretinal sources. There has been little impetus to follow up these computations with experimental observations, because the vertical disparities that normally occur between the images in the two eyes have always been regarded as being too small to be of significance for visual perception and because experiments have consistently shown that our conscious appreciation of egocentric distance is rather crude and unreliable. Nevertheless, the veridicality of stereoscopic depth constancy indicates that accurate distance information is available to the visual system and that the information about egocentric distance and horizontal disparity are processed together so as to continually recalibrate the horizontal disparity values for different absolute distances. Computations show that the recalibration can be based directly on vertical disparities without the need for any intervening estimates of absolute distance. This may partly explain the relative crudity of our conscious appreciation of egocentric distance. From published data it has been possible to calculate the magnitude of the vertical disparities that the human visual system must be able to discriminate in order for depth constancy to have the observed level of veridicality. From published data on the induced effect it has also been possible to calculate the threshold values for the detection of vertical disparities by the visual system. These threshold values are smaller than those needed to provide for the recalibration of the horizontal disparities in the interests of veridical depth constancy. An outline is given of the known properties of the binocularly activated cells in the striate cortex that are able to discriminate and assess the vertical disparities. Experiments are proposed that should validate, or otherwise, the concepts put forward in this paper. PMID- 2573900 TI - [Toxic glomerulonephritis]. AB - Most glomerulopathies are immunologically-mediated. Their pathogenesis is now better understood. The role of cell-mediated immunity has recently been envisaged. The role of circulating antibodies now seems to be more important than that of circulating immune complexes. Antibodies may recognize structural or "planted" antigens in the kidney, the latter being non-renal molecules that may bind renal structures for non-immune reasons. The linear or granular pattern observed at immunofluorescence depends upon the regular or irregular distribution of the antigen. In susceptible individuals, various toxins (heavy metals such as mercury or gold, drugs with an SH group, non-steroidal anti-inflammatory agents) may induce an immune glomerulopathy. It has recently been shown that Brown-Norway rats exposed to one of the above-mentioned agents develop anti-self class II T lymphocytes that are responsible for a polyclonal activation of B cells. Among the various autoantibodies that are produced, some have a nephritogenic potential. Other drugs are responsible for glomerular lesions due to a direct toxic effect of the compound. Doxorubicin induces a nephrotic syndrome in the rabbit, while mitomycin induces a haemolytic uraemic syndrome in humans. Finally, drug addiction often leads to glomerulosclerosis. PMID- 2573902 TI - Oscillation period of MEPP frequency at frog neuromuscular junctions is inversely correlated with release efficacy and independent of acute Ca2+ loading. AB - Periodic oscillations in miniature endplate potential (MEPP) frequency have been described at the frog neuromuscular junction. It is assumed that the periodic oscillations in MEPP frequency reflect cytosolic oscillations in intracellular Ca2+ concentration. In the course of a study related to describing the differences between weak and strong neuromuscular junctions by using the post tetanic potentiation of MEPP frequency, we noted periodic oscillations in MEPP frequency in the first few minutes after a tetanus. The period of this oscillation (i.e. the time interval of one complete oscillation cycle) was inversely related to synaptic release efficacy, as measured by quantal content released per 100 microns of nerve terminal length. Junctions of high release efficacy have an oscillation period of 20 s or less whereas the oscillations in weaker junctions have periods of up to 60 s or longer. This relation is very similar during post-tetanic recovery in either a calcium containing Ringer solution or in a zero calcium-EGTA Ringer solution, indicating that external calcium is not necessary to express the phenomenon. We also found that the oscillations are apparent in resting junctions preceding a tetanus and that they are similar in period and show the same inverse relation to synaptic strength. PMID- 2573903 TI - D1 and D2 dopamine receptor interactions with pilocarpine-induced oral activity in rats. AB - To investigate the relationship between dopamine (DA) and acetylcholine (ACh) systems in the control of oral movement, we studied the effects of specific D1 and D2 drugs on vacuous chewing movements induced by the muscarinic ACh agonist, pilocarpine. In previous experiments we found that when given alone, the D1 agonist SKF 38393 increased vacuous chewing and the D1 antagonist SCH 23390 decreased it, while both the D2 agonist LY 171555 (quinpirole) and the D2 antagonist sulpiride decreased vacuous chewing. In the present experiment, the effects of the D1 drugs had similar effects in rats concurrently given pilocarpine. In contrast, the effects of both of the D2 drugs were altered by pilocarpine. Surprisingly, the actions of D2 agonist and antagonist were affected in opposite ways. The effect of sulpiride in reducing oral movement activity was eliminated by pilocarpine, while the effect of LY 171555 in reducing oral movement was enhanced by pilocarpine. PMID- 2573904 TI - Memory facilitation produced by dopamine agonists: role of receptor subtype and mnemonic requirements. AB - The role of dopamine (DA) receptor subtypes in the acquisition of two memory tasks in the 8-arm radial maze was examined. The receptors were manipulated with posttraining, subcutaneous injections of an indirect DA receptor agonist (D amphetamine), a selective D2 receptor agonist (LY171555), and a selective D1 receptor agonist (SKF-38393). On a win-stay task (sensitive to caudate nucleus lesions) a light cue signalled the location of food in 4 randomly selected arms on each trial. Rats were given one trial per day and injected after training on day 5. D-Amphetamine (2.0 mg/kg) and LY171555 (2.0 mg/kg) improved performance relative to controls; however SKF-38393 (1-4 mg/kg) had no effect on the acquisition of win-stay behavior. On a win-shift task (sensitive to fornix/hippocampal lesions) a delay of 18 hr was imposed between the first 4 and second 4 choices; drugs were injected after the first 4 choices. D-Amphetamine (1.0 mg/kg) and LY171555 (2.0 mg/kg) significantly improved retention relative to controls. SKF-38393 (1-4 mg/kg) had no effect on win-shift retention. These results suggest that the memory-improving properties of DA agonists on tasks sensitive to both hippocampal and caudate lesions are mediated by the D2 receptor. PMID- 2573905 TI - Long-term central 5-HT depletions resulting from repeated administration of MDMA enhances the effects of single administration of MDMA on schedule-controlled behavior of rats. AB - The behavioral effect of single administration of +/- 3,4-methylene dioxymethamphetamine (MDMA) on rats performing on the differential-reinforcement of-low-rate 72-second schedule (DRL 72-sec) was compared before and after a period of repeated administration of MDMA known to deplete 5-hydroxytryptamine (5 HT) levels in the brain. Single administration of MDMA decreased reinforcement rate (1, 2, 4, 6 mg/kg) and increased response rate (4,6 mg/kg) of rats performing on the DRL 72-sec schedule. This effect is typical of amphetamines and other psychomotor stimulants. Four weeks after repeated administration of MDMA (6 mg/kg twice daily for 4 days) there was an increase in sensitivity to the effect of single administration of MDMA. Doses of 2, 4 and 6 mg/kg of MDMA resulted in increases in response rate that were significantly greater after repeated MDMA administration than before. Doses of 0.5, 2, and 6 mg/kg of MDMA resulted in decreases of reinforcement rate that were significantly greater after repeated MDMA administration than before. Repeated administration of MDMA resulted in long term depletion of serotonin levels by 30-50% in the amygdala, neostriatum, hippocampus and the frontal cortex. Norepinephrine and dopamine (DA) levels were not significantly different from control in any of the brain regions analyzed. The behavioral and neurochemical results suggest that serotonergic neurons normally exert an inhibitory action upon the psychomotor stimulant effects of MDMA. Since the psychomotor stimulant effects of amphetamines appear to be mediated primarily by the dopamine system, these results provide evidence that 5 HT and DA may represent opposing systems in the DRL schedule-controlled behavior. PMID- 2573906 TI - Pharmacology of calcium-induced long-term potentiation in rat hippocampal slices. AB - A transient increase (10 min) in extracellular calcium concentration (4 mM) causes a long-lasting (greater than 2 hr) enhancement of population spike responses evoked by radiatum fibers to CA1 pyramidal neurons in rat hippocampal slices. This phenomenon is similar to tetanic long-term potentiation (LTP), and is also related to memory processes. The influence of various drugs was investigated on calcium-induced LTP. The NMDA antagonist 2 amino-5 phosphonopentanoic acid (AP5; 100 microM) was able to prevent the calcium-induced LTP, while atropine sulphate (10 microM), propranolol hydrochloride (10 microM) and verapamil hydrochloride (100 microM) were ineffective. The results suggest an involvement of the NMDA receptor in the development of calcium-induced LTP. PMID- 2573907 TI - Celiprolol, a compound which attenuates myocardial acidosis and improves regional segmental function following ischemia in dogs: a comparison with propranolol. AB - Celiprolol, propranolol or saline were administered to separate groups (n = 5-6) of anesthetized dogs in which a critical stenosis was applied to the circumflex coronary artery for 90 min and then reperfused for 30 min. Test drugs were administered at 30 min poststenosis and the effects on pH, regional function and endocardiogram were monitored. A reduction in coronary flow of 54 +/- 2% (n = 27) yielded marked increases in hydrogen ion concentration (H+) of 17 +/- 2 X 10(-8) and ischemic endocardial ST segment of 6 +/- 1 mV while ischemic segmental shortening decreased 75 +/- 9%. Heart rate, arterial pressure and normal regional function were not altered. Celiprolol 0.1 and 1 mg/kg, i.v., reversed the alterations in H+ and ischemic ST segment to prestenosis values while improving ischemic segmental shortening 20 and 38%, respectively, and not affecting heart rate. Propranolol 0.1 and 1 mg/kg, i.v., reversed the alterations in H+ and ischemic ST segment to prestenosis values while further decreasing ischemic segmental shortening 66 and 30%, respectively. Upon reperfusion, ischemic segmental shortening returned to prestenosis values in the group treated with celiprolol 1 mg/kg, i.v., while the propranolol- and saline-treated groups further decreased. It is concluded that celiprolol is efficacious in normalizing myocardial function and ischemia-induced electrophysiological changes following coronary artery stenosis. PMID- 2573908 TI - Effects of hydralazine on guanosine cyclic 3', 5'-monophosphate levels in rat aorta. AB - The mechanism of the vasodilator effect of hydralazine on isolated rat aorta was studied. Results demonstrated that the vasodilator effect of hydralazine was greater on intact aortas than on endothelium-denuded preparations, particularly at low concentrations of between 0.1 mM and 0.5 mM. In addition, hydralazine did not have any effect on cyclic GMP levels. We also found that methylene blue, an inhibitor of guanylate cyclase, completely abolished the vasorelaxant action of nitroglycerin but not that of hydralazine. These results indicate that the vasodilator effect of hydralazine was not due to elevating the cyclic GMP levels. On the other hand, hydralazine significantly inhibited both the contractions induced by norepinephrine and/or high-potassium. In conclusion, a part of the vasodilator effect of hydralazine seems to depend on the integrity of the vascular endothelium. However, this vasodilator effect was not associated with any elevation in cyclic GMP level. Thus, the direct vasodilator action of hydralazine may be related to its interference with the movement and/or translocation of calcium across the cell membrane. PMID- 2573909 TI - Control of phrenic nerve activity and blood pressure by the medullary raphe nuclei in cats. AB - Electrical and chemical stimulation methods were used to determine the topographic organization of the medullary raphe nuclei (MRN) in controlling the systemic arterial blood pressure (BP) and phrenic nerve activities (PNA). Decerebrated, unanesthetized and bilateral vagotomized cats were used. Effective points in the MRN were systematically explored with constant current stimulation. We found stimulation of the rostral MRN produced a decrease in PNA amplitude and increase in BP and PNA frequency. Stimulation of the caudal MRN produced increases in BP and the amplitude and frequency of PNA. Microinjection of glutamate solution into the caudal or the rostral MRN points produced qualitatively similar results. Thus, we concluded that the caudal MRN neurons had excitatory connections whereas the rostral MRN neurons had excitatory and inhibitory connections to the cardiovascular preganglionic neurons and the phrenic nerve motoneurons. PMID- 2573910 TI - Presynaptic modulation of transmitter release by nicotinic receptors. PMID- 2573911 TI - Primary cholinergic and indirect dopaminergic mediation of behavioural effects of nicotine. PMID- 2573912 TI - Studies of protective actions of nicotine on neuronal and vascular functions in the brain of rats: comparison between sympathetic noradrenergic and mesostriatal dopaminergic fiber systems, and the effect of a dopamine agonist. AB - Neuroprotective and possible trophic actions of nicotine were studied in two types of experimental models: (1) one in which the meso-striatal dopamine system was subjected to partial hemitransection, and regional glucose utilization (using 2-[3H]deoxyglucose) and blood flow (using [14C]iodoantipyrine) were measured by computer-assisted quantitative autoradiography based on a double-isotope technique; and (2) another where the sympathetic cranial nervous system supplying the brain vasculature was subjected to decentralization, axotomy, and partial or complete ganglionectomy, and the neuronal survival and fiber regeneration were elucidated by fluorescence histochemistry of noradrenaline, tyrosine hydroxylase, and neuropeptide Y. Continuous nicotine infusion for 4 weeks failed to significantly affect the neuronal response to the surgical interference of the sympathetic noradrenergic system. The same nicotine treatment for 2 weeks significantly improved glucose utilization and blood flow in caudate-putamen on the side in which the meso-striatal dopamine system had been transected, thus eliminating the 16% side-to-side asymmetry in the metabolism caused by the axotomy. The dopamine agonist, EMD 23,448, was without significant effect on this asymmetry. The hemitransection produced marked reduction in metabolism and flow also in the ventro-lateral thalamus. In substantia nigra, glucose utilization was markedly elevated--perhaps as a consequence of a regenerative increase in protein synthesis--opposite to a considerable reduction in nigral blood flow. Little or no effect of the hemitransection was seen in hippocampus or nucleus accumbens. In neither of these four regions did nicotine (or EMD 23,448) have any overt influence on glucose metabolism or blood flow. It is concluded that nicotine, mainly through its protective action on the meso-striatal dopaminergic system, is able to improve striatal glucose utilization and associated blood flow, probably reflecting a tendency to amelioration of neurotransmission function of surviving terminals belonging to the nigro-striatal dopamine neurons. PMID- 2573913 TI - Single cell studies of the actions of agonists and antagonists on nicotinic receptors of the central nervous system. PMID- 2573914 TI - Hemorrhagic fever with renal syndrome. AB - Hantaviruses, the causative agents of HFRS, have become more widely recognized. Epidemiologic evidence indicates that these pathogens are distributed worldwide. People who come into close contact with infected rodents in urban, rural and laboratory environments are at particular risk. Transmission to man occurs mainly via the respiratory tract. The epidemiology of the hantaviruses is intimately linked to the ecology of their principal vertebrate hosts. Four distinct viruses are now recognized within the hantavirus genus and that number is likely to increase to six very soon; however, further investigations are necessary. Much more work is still needed before we fully understand the wide spectrum of clinical signs and symptoms of HFRS as well as the pathogenicity of the different viruses in the hantavirus genus of the Bunyaviridae family. HFRS is difficult to diagnose on clinical grounds alone and serological evidence is often needed. A fourfold rise in IgG antibody titer in a 1-week interval, and the presence of the IgM type of antibodies against hantaviruses are good evidence for an acute hantavirus infection. Physicians should be alert for HFRS each time they deal with patients with acute febrile flu-like illness, renal failure of unknown origin and sometimes hepatic dysfunction. Especially the mild form of HFRS is difficult to diagnose. Acute onset, headache, fever, increased serum creatinine, proteinuria and polyuria are signs and symptoms compatible with a mild form of HFRS. Differential diagnosis should be considered for the following diseases in the endemic areas of HFRS: acute renal failure, hemorrhagic scarlet fever, acute abdomen, leptospirosis, scrub typhus, murine typhus, spotted fevers, non-A, non-B hepatitis, Colorado tick fever, septicemia, dengue, heartstroke and DIC. Treatment of HFRS is mainly supportive. Recently, however, treatment of HFRS patients with ribavirin in China and Korea, within 7 days after onset of fever, resulted in a reduced mortality as well as shortened course of illness. PMID- 2573915 TI - [Cell cycle regulation in M-phase]. PMID- 2573917 TI - [Interdisciplinary aspects of the treatment of hypertension]. PMID- 2573916 TI - [Diagnosis of arterial hypertension]. PMID- 2573918 TI - Selective histamine blockade in childhood asthma; the effect of terfenadine on resting bronchial tone and exercise induced bronchoconstriction. AB - The orally active histamine H1 blocker terfenadine was investigated for its effect on resting bronchial tone and exercise induced bronchoconstriction in 20 asthmatic children by a double blind placebo controlled study. Terfenadine produced acute broncholidation with an average 32% improvement in FEV1 by 3 h. After exercise challenge terfenadine partially inhibited exercise induced bronchoconstriction. The maximum fall in PEFR after exercise was reduced from 32% after placebo to 21.5% after terfenadine. These results suggest that asthmatic children have background resting 'histamine tone' that is reversible by histamine H1 blockage. PMID- 2573919 TI - Management of asthma in general practice. AB - An audit of the management of asthma in two large general practices has been undertaken. The overall level of therapy prescribed was, in general, related to both the objective severity of the patients' asthma and the extent of symptoms. However, many individual patients received sub-optimal therapy. Prophylactic inhaled beta agonists were used infrequently. Inhaled steroids were prescribed to only one third of the patients and to less than half of severely affected patients. The results suggest that this group of adult asthmatics were relatively under treated in general practice, but a prospective study with proven compliance is necessary to confirm this. PMID- 2573921 TI - [Current problems in the therapeutic use of theophylline]. PMID- 2573920 TI - [The diagnosis of drug-induced hepatopathies]. AB - The present paper reports on the recent possibilities of diagnosis in iatrogenic diseases of the liver. The iatrogenic diseases of the liver are diagnosed with difficulty since no clinical, biological and histological pathognomonic criteria are known. The hepatic lesions appearing after drug intake are extremely various, including: hepatocellular necrosis, cholestasis, hepatic granuloma, perisinusoid fibrosis, steatosis, pseudoalcoholic lesions, vascular lesions and hepatic tumours. Clinical criteria, very important for the diagnosis, are based on: appearance of the disease, regression of symptomatology when the treatment is interrupted and recurrence when it is administered again. The biological criteria are not specific but they may indicate the drug etiology of a liver disease. Increase in the serum concentration of aminotransferases might be the only biochemical disturbance and it might be overlooked if not systematically investigated. The immunological tests suggested for diagnosis of iatrogenic liver diseases are numerous but only a few of them are specific. Histological criteria cannot identify, for sure, the drug etiology of the lesions. They may suggest the possibility of a drug induced cause and may help in establishment of a correct diagnosis. PMID- 2573922 TI - [Bronchopulmonary cancer in persons under 40 of age]. PMID- 2573924 TI - [Campylobacter pylori infection in chronic gastritis and gastric cancer]. AB - The present paper analyses, in correlation with histologic lesions, the incidence of infection with Campylobacter pylori (CP) in patients with chronic gastritis, operated stomach, gastric neoplasm and healthy subjects. In chronic gastritis, incidence of infection with CP is 71.6%, significantly increased in comparison with the controls, in whom the positive percentage is 37.5. The most of the patients in this group suffered from superficial chronic gastritis. In the group with operated stomach, the CP infection included 48% of the patients studied; all had superficial chronic gastritis. In gastric cancer, CP bacteria appeared in 50% of cases. CP may represent a pathogenetic ring that starts the gastric process. PMID- 2573923 TI - [The effects of nitroglycerin administered intravenously in acute myocardial ischemia]. AB - The effects of i.v. nitroglycerin were studied by ECG and enzymatically in 16 patients (mean age 57.9 +/- 1.4 years) (NTG) in comparison with a control lot (c) of 17 patients (mean age 62.7 +/- 2.1 years) treated with dipyridamole and/or nifedipine (N), admitted in the first 4-10 hours after the onset of the first symptoms. The patients with heart failure and those with Q waves and CPK or LDH values greater than 2 x n were not admitted. NTG was administered in doses of 20 micrograms--60 microgram/hour for 24-96 hours and systolic AT (s) was kept under 10% of the basic values but not under 100 mmHg. Myocardial infarction appeared in 9 N-treated patients (54.86%) and 11 controls (58.25%) (p = 0.07). The size of myocardial necrosis was reduced in the N-treated patients. Peak serum CPK levels had considerably less increases in N (from 72.9 U to 73.4 U) (p greater than 00.5) versus C from 34.2 U to 364.5 U) (p less than 0.001). The sum of segmentary depression failed from 9.13 mm to 3.19 mm (p less than 0.05) in N, whereas in C the decrease was not significant (6.12 mm as against 9.38 mm; p greater than 0.05). The evolution was severe in C, as the angina crises (14 cases versus 2 cases, p less than 0.01) and the extension of the infarction (8 cases versus, 0; p 0.05) less than 0.05) appeared more frequently than in N. Only two patients in C died (p less than 0.05). Therefore, i.v. NTG administration in small doses in acute myocardial infarction leads to immediate disappearance of the anginal pain, lowers the extent of the myocardial necrosis and improves the clinical evolution. PMID- 2573925 TI - [The dumping syndrome: its risk factors and evolution]. AB - The paper reports on a comparative study of 24 cases of operated gastroduodenal ulcer, in which the dumping syndrome appeared at a short interval after the operation, and of a control lot of patients operated for ulcerous disease without complications. This study shows that the incidence of the dumping syndrome in the ulcerous patients operated is of 12.83%, predominant in gastric ulcer (15%). Among the factors favouring the development of the dumping syndrome, mention is made of subtotal gastric resection with Billroth II anastomoses, early surgical indication, and pre-existing neuroendocrine disturbances. Although the dumping syndrome causes weight loss, its evolution in time is benign and improving after 3 years, surveillance. PMID- 2573926 TI - [Characteristics of current treatment of urinary infections in clinical practice]. PMID- 2573927 TI - [Acute lymphoblastic Ph1-positive leukemia with a hybrid phenotype. The significance of the Ph1 anomaly and the hybrid phenotype]. AB - The present paper reports on a case of acute lymphoblastic leukemia with t(9q+; 22q-) and surface markers specific of the B and T lymphoid line (hybrid phenotype). The fundamental (genotypical and phenotypical) and practical aspects (nosologic and therapeutical aspects) of this particular subtype of Ph1-positive acute leukemia and hybrid phenotype are discussed. PMID- 2573928 TI - [Hepatic tuberculosis]. AB - Hepatic tuberculosis is well-known anatomicopathologically, consequent to necropsy and liver biopsy puncture, but the clinical aspect when the signs of the pulmonary localization are not dominant, may be similar to other "masked" affections, hepatic or extrahepatic, more often chronic hepatitis, hepatic tumours or prolonged febrile syndrome with many etiologic possibilities. The past history of the patients is very important as it confirms the observation of the histopathologic and possibly bacteriological examinations of a liver fragment. Three cases presumptively clinically diagnosed are presented and histopathologically verified for confirmation of the facts illustrated. PMID- 2573929 TI - [Current developments in antirheumatic medication. II. Metabolic and cartilage protective medication]. PMID- 2573930 TI - [Diabetic nephropathy. I]. AB - The paper reports on the present knowledge in the diabetic nephrosis (DN) with emphasis on three problems: evolution by stages, possibility of detecting the diabetics with high risk of developing clinic DN, and the prophylactic and therapeutic measures required for a resting or, at least, for slowing down the evolution of this complication. The functional changes, manifested mainly by increasing the glomerular filtration, are detected in many insulin-dependent diabetics even from the diagnosis of the disease and they are predictive elements for the clinic DN; that is the reason why they must be reduced by a correct and prolonged control. Microalbuminuria, defined as a urinary excretion of albumin of less than 250 mg/24 hours (18-74 micrograms/minute) is another characteristic of the precocious stages of the DN. Exceeding the above-mentioned values means the transition to the clinic DN, that subsequently progresses in all cases. The paper also describes the characteristics of the clinic DN and the therapeutic ways both in the incipient and clinic DN, with emphasis on their features in the diabetes mellitus and its complications. PMID- 2573931 TI - [The early exercise test following an acute myocardial infarct in selecting patients with an increased coronary risk]. PMID- 2573932 TI - [Possibilities for the use of the leukocyte migration inhibition test in clinical practice]. PMID- 2573933 TI - [Asymptomatic biliary lithiasis]. PMID- 2573934 TI - [Current developments in immunopathology of the intestines]. PMID- 2573935 TI - [Diabetic neuropathy: clinical and electrophysiologic results of the use of a plant extract rich in myoinositol]. PMID- 2573936 TI - [The incidence of chronic hepatitis in patients with a stomach resected in gastroduodenal ulcer]. PMID- 2573937 TI - [Treatment with antidepressants in biliary dyskinesia and irritable colon]. PMID- 2573938 TI - [The evaluation of colchicine treatment in certain forms of moderately active chronic hepatitis]. PMID- 2573939 TI - [Glucose regulation in diabetes mellitus (the Somogyi effect and the "dawn" phenomenon)]. PMID- 2573940 TI - [Diabetic nephropathy. IIa]. PMID- 2573941 TI - [Hypertensive cardiopathy--an adaptive or a pathologic phenomenon?]. AB - The hypertensive cardiopathy is a controversial entity as regards the physiopathological mechanisms and clinical aspects. Defined as the hypertrophy of the left ventricle, secondary to the permanent tension increases, it accompanies not only the severe forms of arterial hypertension but also the medium and mild ones. In the authors' opinion, based on numerous experimental and clinical studies, the main factor that initiates the myocardial hypertrophy is the increased parietal tension (hemodynamic hypertrophy). The natural evolution is progressive, the myocardial hypertrophy initially adaptive becomes pathological and the cardiac performance is affected gradually, first in its diastolic and then in its systolic component, up to the final stage of congestive cardiac insufficiency. The structural changes of the myocardial fibre also document the adaptive and pathological hypertrophy, the alteration of the myocardial contractility consisting in the difficulty of transforming the chemical energy into mechanical work. The clinical aspects show an incipient myocardial hypertrophy, considered adaptive, since the cardiac performance is normal; an important hypertrophy affecting the diastolic component and the hypertrophy with dilatation that affects the overall performance. Of the evaluation methods, the echocardiography is the most accurate one in quantifying hypertrophy, evaluation of the cardiac performance and possibility of detecting several characteristic aspects of the hypertensive cardiopathy. The transition moment from the adaptive hypertrophy to the pathological hypertrophy cannot be exactly established but it is documented that the hemodynamic and nonhemodynamic hypertrophy is a supplementary cardiovascular risk factor. PMID- 2573942 TI - [Intracellular microinjection--a useful method in biomedical research]. PMID- 2573943 TI - [Anatomo-clinical correlations in pulmonary thromboembolism (observations on 258 cases)]. AB - The authors studied 258 cases of deaths due to lung thromboembolization, totalling 13% of the necropsies. The patients' age ranged within 17-89 years, with the highest frequency between 70 and 75 years. The female sex was prevalent (56% of cases). According to the magnitude of the vessel occluded, the thromboembolisms were divided into: massive (35%), medium (50%) and small (15%). The lung infarctions were present in 42% of the cases investigated. The formation site of the thrombi was exactly established in 24% of the cases; the right side of the heart, the lung, the deep veins of the shank, periprostatic venous plexus, vena cava inferior, iliac veins. The lung thromboembolization was favoured by the cardiovascular affections (38.5%), neurologic affections (27%), malignant tumours (11.5%), postsurgery status (9%), lung diseases (9%), coma (3.5%), prostate affections (1.5%). The clinical diagnosis was established in 22% of cases. In the rest of them, thromboembolization was hidden or simulated by the coexisting affection. PMID- 2573944 TI - [The anticoagulant treatment of patients with heart valve prostheses]. PMID- 2573946 TI - [The effect of withdrawal on humoral and cellular immune changes in alcoholic hepatopathy]. AB - In 24 patients with alcoholic hepatopathy (HA) the long-time (two years) influence--induced by the alcohol withdrawal (16 patients) or continuation (8 patients)--on the immunological, humoral and cellular modifications was followed. An important improvement of the immunological disorders, mainly the cellular ones, in the weaned patients was noticed: the increase of T lymphocytes with "reestablishment" of the T lymphocyte subsets equilibrium by increasing the T1 subpopulation--s/c: the decrease of the active T lymphocytes and of the B lymphocyte population. These changes led to humoral immune improvement. In the non-withdrawal patients, the immunological disorders continued and became even worse. PMID- 2573947 TI - [Is an esophageal transit scintigram necessary in patients with gastroesophageal reflux?]. AB - Efficient esophageal clearance has an important defence role in the pathogenesis of the gastroesophageal reflux disease (GERD). Many GERD patients have esophageal disturbances associated with or secondary to reflux, producing delayed clearance. This delay exposes the esophageal mucosa to the reflux acid content. To determine esophageal transit we scanned the esophageal transit of a 15 ml bolus containing colloidal 300/cCi 99m Tc. The esophageal transit was calculated in seconds according to formula E.T. = T 1/2 x 5. The study included 74 GERD patients. The following investigations were carried out in all the cases: esophageal X-ray, GER scintigram, endoscopy, esophageal biopsy, Bernstein test and esophageal transit scintigram. Endoscopy revealed lesions of the esophagus (of 1st, 2nd and 3rd degree) in 39 patients, Barrett syndrome in 8 cases and normal in 27. Esophageal transit scanning was normal in 18 cases (24%), and prolonged in 56 cases (76%). Only 7 (39%) of the 18 patients with a normal transit presented lesions of the mucosa, the latter being more frequent in patients with a prolonged transit, i.e. 40 of 56 patients (71.5%). The mean value of the transit in different degrees of esophagitis (I, II, III) and Barrett syndrome were: 12.73 +/- 5.36; 13.30 +/- 7.90; 10.35 +/- 5.78; 17.25 +/- 11.17. In conclusion esophageal transit scanning is a useful test in GERD patients as it has a prognostic value. A prolonged esophageal transit is frequently associated with lesions, the more severe the slower is the transit. Moreover the test may indicate certain drugs acting upon the esophageal motor disturbances. PMID- 2573945 TI - [The cholestasis syndrome. A critical evaluation of an exploratory strategy]. PMID- 2573948 TI - [Clinical and morphopathologic study of lupus vasculitis]. PMID- 2573949 TI - [Elevated eosinophilias (30-90%) and their place among parasitic and nonparasitic diseases (personal observations)]. AB - The 237 cases of hypereosinophilia studied, ranging from 30 to 90% showed that, except for two cases that were not confirmed by the repeated blood leukograms, 22 had a parasitic origin, 6 appeared in certain pathologic states, or various severe diseases, 5 had an undetermined etiology and 2 were off the record. The authors draws the attention on the value of various methods of investigation which he used especially in the diagnosis of the parasitic diseases met with. PMID- 2573950 TI - [Enzymatic aspects of severely acute diarrheal disease in infants]. AB - Acute dehydration of diarrhoea of the breast-fed child is a type of hypovolemic shock which, by its severity and frequency is one of the major emergencies of pediatrics. The liver, as the site of many metabolic processes is mainly affected in the frame of the multiple-tissue involvement which is characteristic for this condition. The authors carried out a study of several enzymes (total LDH and the thermally-labile fraction, gluthamatdehydrogenase, gammaglutamyltranspeptidase, acid and alkaline phosphatase) in 27 breast-fed children. Determination of the enzymes was done on hospitalization, and 10 days later. The study showed that all the enzymes that were evaluated had moderate increases in 40.7-71% of all cases. The increases were significant (p less than 0.001). After 10 days the enzyme titers showed a tendency to become normal again, and slightly higher values were found in only 15-12% of all cases. Minimal hepatopathy can be considered, associated to the clinico-biological evolution of this severe conditions of the breast-fed child pathology. PMID- 2573951 TI - Current concepts in brain ischemia. A review. PMID- 2573952 TI - T-cell response to phorbol ester PMA and calcium ionophore A23187 in Down's syndrome. AB - The proliferative response of purified T cells to anti-CD2 monoclonal antibodies (T112 plus T113) was found to be markedly reduced in 12 subjects with Down's syndrome (DS). The addition of phorbol ester PMA, which activates Ca2+/phospholipid-dependent enzyme protein kinase C, or calcium ionophore A23187, which increases intracytosolic free Ca2+ concentration, enhanced, but did not normalize, the defective anti-CD2-mediated T-cell mitogenesis. In contrast, the proliferation of resting lymphocytes from trisomic patients was comparable to that of the control cells when PMA and A23187 were used as co-blastogenic reagents. Because PMA and A23187 together bypass the early activation pathways and promote T-cell growth through the direct induction of membrane interleukin 2 (IL-2) receptor expression and IL-2 synthesis and secretion, it could reasonably be hypothesized that the faulty DS T-cell activation induced by antigen or mitogen is due to a deranged transmembrane signal transduction, rather than a defect in the later intracellular events. PMID- 2573953 TI - A new DNA marker tightly linked to the fragile X locus (FRAXA). AB - The fragile X syndrome is the most common cause of familial mental retardation. Genetic counseling and gene isolation are hampered by a lack of DNA markers close to the disease locus. Two somatic cell hybrids that each contain a human X chromosome with a breakpoint close to the fragile X locus have been characterized. A new DNA marker (DXS296) lies between the chromosome breakpoints and is the closest marker to the fragile X locus yet reported. The Hunter syndrome gene, which causes iduronate sulfatase deficiency, is located at the X chromosome breakpoint that is distal to this new marker, thus localizing the Hunter gene distal to the fragile X locus. PMID- 2573955 TI - The surgical treatment of gastrinoma in MEN I syndrome patients. AB - The pancreatic component of the multiple endocrine neoplasia type I (MEN I) syndrome is a difficult and controversial problem because the entire endocrine pancreas is diffusely involved with varying degrees of islet-cell hyperplasia, microadenomatosis, and nesidioblastosis. In addition, in patients with functional syndromes, islet-cell tumors usually develop, and these may or may not be malignant. Because of the presumed inability to alleviate or cure the Zollinger Ellison syndrome (ZES) in MEN patients, total gastrectomy was the treatment of choice before the introduction of H2 antagonists and omeprazole. At present, many physicians and surgeons consider H2 antagonists the best treatment and advise pancreatic exploration only when a gross pancreatic tumor is demonstrated on imaging studies. During the past 10 years we have studied all MEN I patients with ZES without hepatic metastases or gross pancreatic tumors using percutaneous transhepatic selective venous gastrin samplings. Two patterns of gastrin secretion were identified: (1) diffuse from multiple pancreatic sites and (2) localized regional secretion. Four patients from the latter group were selected for attempted surgical "cure" without gastrectomy or total pancreatectomy. Two additional patients are included who had resection of gastrinomas and have maintained basal serum gastrin levels within the normal range for extended periods. The follow-up on these patients ranges from 5 months to 12 years. All six patients have normal basal gastrin values, and those with remaining stomachs require no drug therapy. PMID- 2573954 TI - Hypergastrinemia after vagotomy is not associated with decreased gastric somatostatin. AB - Although hypergastrinemia occurs after vagotomy, the mechanisms responsible are not understood. Somatostatin (SRIF) is a peptide that inhibits gastrin release, is present within the gastric fundus and antrum, and is under vagal control. In this study we have investigated the hypothesis that hypergastrinemia is associated with a decrease in gastric SRIF. We examined tissue levels of SRIF in gastric mucosa and muscle wall in rabbits undergoing vagotomy and pyloroplasty compared with sham-operated controls. We also compared the release of SRIF from gastric glands in response to vasoactive intestinal peptide and calcitonin gene related peptide. Vagotomy resulted in an increase in gastrin compared with controls in both antrum (1062 +/- 176 pmol/gm vs 484 +/- 48 pmol/gm) and plasma (236 +/- 72 pmol/L vs 29 +/- 4 pmol/L; p less than 0.05). This was accompanied by an increase in the number of gastrin cells (25 +/- 4 vs 9 +/- 3; p less than 0.05). No significant differences were observed in gastric SRIF levels in either the fundus or antrum (p greater than 0.5). In addition, there were no differences in the release of SRIF from gastric glands of vagotomized animals compared with controls in response to vasoactive intestinal peptide and calcitonin gene-related peptide (p greater than 0.5). These data suggest that the elevations in plasma and antral gastrin levels after vagotomy are not a result of alterations in gastric SRIF. PMID- 2573956 TI - Selective surgical management of correctable hypergastrinemia. AB - Sixty patients with surgically correctable hypergastrinemia were treated between 1960 and 1988. Provocative testing was used when available to select appropriate operations. Sources of hypergastrinemia included antral G cell hyperplasia (AGCH) (17), pancreatic gastrinomas (14), duodenal gastrinomas (11), multiple gastrinomas in patients with type I multiple endocrine neoplasia (MEN I) (five), lymph node gastrinomas (four), and the source not found in nine patients. Eugastrinemia was achieved by resection in 17 of 17 patients with AGCH, nine of 11 patients with duodenal gastrinomas, three of four patients with lymph node gastrinomas, zero of 14 patients with pancreatic gastrinomas, zero of five patients with MEN I, and zero of nine patients in whom the source was not found. Hepatic metastases developed in 11 patients with pancreatic gastrinomas, two patients with MEN I, one patient with duodenal gastrinomas, and one patient with lymph node gastrinomas. One patient in whom the source of the hypergastrinemia was not found developed hepatic metastases, and seven required total gastrectomy. This experience suggests the following: (1) that patients with AGCH, duodenal gastrinomas, or lymph node gastrinomas can usually be rendered eugastrinemic by resection; (2) that patients with pancreatic gastrinomas, whether sporadic or familial (MEN I), are rarely cured by resection and frequently develop hepatic metastases; and (3) that patients in whom the source of the hypergastrinemia is not identified and removed frequently require total gastrectomy, but antroduodenectomy should be considered because it may uncover an occult duodenal microneurogastrinoma or may correct AGCH. PMID- 2573957 TI - Management of islet cell tumors in patients with multiple endocrine neoplasia: a prospective study. AB - As part of a study to manage islet cell tumors in patients with multiple endocrine neoplasia (MEN), patients with MEN I and Zollinger-Ellison syndrome (ZES) underwent surgery if a pancreatic islet cell tumor was identified on imaging studies. Patients with MEN I and either insulinoma or vasoactive intestinal polypeptide tumor (VIPoma) underwent surgery whether or not a tumor was identified. Each patient underwent preoperative portal venous sampling (PVS). Nine patients with MEN I and one with MEN II underwent surgery; seven had ZES, one had insulinoma, one had VIPoma, and one had both insulinoma and ZES. Eight of the nine patients with MEN I had an identifiable hormone gradient on PVS. Islet cell tumors were removed from the pancreas of each patient; two patients also had duodenal wall tumors, and three patients had malignant islet cell tumors. No patient with ZES and MEN I was cured of ZES despite the fact that islet cell tumor was removed from the region of the gastrin gradient in five of six patients. The single patient with MEN II and ZES and the three additional patients with MEN I and either insulinoma or VIPoma were cured by islet cell tumor resection. The results indicate that islet cell tumors in patients with MEN I can be both extrapancreatic and malignant. In patients with MEN I and ZES, ZES cannot be cured by tumor resection, and PVS cannot be used to select patients for curative surgery. It appears that gastrinoma in patients with MEN II, as well as either insulinoma or VIPoma in patients with MEN I, can be cured by islet cell tumor resection. PMID- 2573959 TI - [Isolation of hemopoietic precursor cells by sequential leukocytapheresis and their clinical use]. PMID- 2573958 TI - H2-receptor blockade induces peptide YY and enteroglucagon-secreting gastric carcinoids in mastomys. AB - Gastric carcinoid tumor formation has been reported with prolonged achlorhydria in both animals and human beings. The hypothesis in this study was that the ablation of parietal cell function in an animal (mastomys) genetically predisposed to gastric neuroendocrine neoplasia would promote and accelerate tumor formation. Loxtidine, an irreversible H2-receptor blocker, was administered at 1 mg/kg/day in drinking water for 4 months to young mastomys (n = 16). After 4 months of treatment, 14 of 16 animals had gastric carcinoids compared with 0 of 16 young control animals and 4 of 16 older control animals. Ultrastructurally, these tumors were characterized by the presence of neurosecretory granules. Serum gastrin levels were elevated (230 +/- 40 pmol/L) in loxtidine-treated animals compared with control animals (26 +/- 8 pmol/L) (p less than 0.05). In addition, both peptide YY (620 +/- 160 pmol/L) and enteroglucagon (500 +/- 147 pmol/L) were significantly elevated compared with control groups (p less than 0.05). Similarly, in tumor tissue, peptide YY (676 +/- 152 pmol/gm) and enteroglucagon (551 +/- 164 pmol/gm) were found in large quantities, whereas gastrin was undetectable. These observations provide substantial support for the possible pathophysiologic role of gut peptides, particularly gastrin, in the generation of endocrine neoplasia. The advent of endocrine tumors after inhibition of a gut secretory cell (parietal) may be of considerable significance in understanding the genesis of endocrine neoplasia. Whether the drug acts as a neoplastic promoter of enterochromaffin-like cells or the tumor development is related to elevation of peptides such as gastrin cannot be established in this study. Long term H2-receptor blockade with new potent, irreversible agents as an alternative to surgery may have potential grave implications that require careful consideration. PMID- 2573960 TI - [Humoral regulators of erythropoiesis during the body's adaptation to chronic respiratory insufficiency]. AB - It was revealed during examination of 91 patients with chronic nonspecific lung diseases for the blood titers of erythropoietin, erythrocytic chalones, erythrocyte hemolysis products and medium molecules that in the stage I respiratory failure, there were no material changes in the titres of erythrocyte hemolysis products, erythropoietin and medium molecules, whereas the titer of erythrocytic chalones appeared to be high. Stage III of the disease was also marked by no changes in the titer of erythrocyte hemolysis products. However, the titers of erythropoietin and medium molecules rose whereas the titer of erythrocytic chalones was reduced. In the stage II respiratory failure, the above enumerated factors of humoral regulation of erythropoiesis appeared similar to those in stage I. These data evidence that different compensatory reactions to hypoxia associated with chronic respiratory failure are implicated at the molecular, cellular and systemic levels. The up-to-date treatment and diagnostic process is not feasible, provided these reactions are not taken into consideration. PMID- 2573961 TI - [Neurohumoral mechanisms regulating renal blood circulation in patients with hypertension undergoing captopril block of angiotensin-converting enzyme]. AB - A study was made of the effect of a single intake of captopril on the neurohumoral mechanisms of the regulation of renal circulation in 25 patients with essential hypertension (EH). Captopril induced an increase of the effective renal blood flow (ERB) and of the effective renal plasma flow along with a considerable lowering of the renal vascular resistance. No relationship was found between these changes and the time course of changes in the activity of plasma renin, aldosterone and vasopressin concentration in blood plasma. The relationship was established between the changes in the ERB and the time course of changes in baroreceptor sensitivity that significantly increased in the majority of EH patients under the effect of captopril. PMID- 2573962 TI - [Bronchomotor effects of cardioselective beta 1-adrenoblockaders in patients with bronchial asthma]. AB - A study was made of the effect of beta 1-adrenoblockers (metoprolol, atenolol, talinolol) on bronchial patency in patients with concomitant bronchial asthma (BA) under acute drug use and in the course of continuous therapy. Broncho obstructive complications occurring in part of the patients were not associated with the disease gravity or with the pathogenetic variety of BA. Besides, they were not associated with the drug dose either (use was made of the mean therapeutic dosage range). The rate and the intensity of bronchial patency abnormalities occurring in the course of the continuous treatment with beta 1 adrenoblockers (with metoprolol, in particular) depended on the initial status of the adrenergic regulation of the body. Significant disorders of bronchial patency including clinically marked ones were naturally observed with initially low excretion of cAMP in the morning portion of urine (less than 3 mmole/1). PMID- 2573963 TI - [A comparative evaluation of the efficacy of sucralfate and H2-histamine blockaders in treating peptic ulcer]. AB - Sucralfate (55 patients), cimetidine (25 patients), ranitidine (30 patients), antacids and cholinolytics (32 patients) were studied and compared in open clinical trials for the efficacy in the treatment of ulcer disease of the stomach and duodenum. During 5 weeks, ulcers completely healed in 100% of the patients given sucralfate (in 94.5% for 4 weeks), in 90% of the patients treated with ranitidine, in 84% on cimetidine, and in 78% of the patients undergoing treatment with antacids and cholinolytics. PMID- 2573964 TI - The mechanism of the action of Minipress. Examinations in hypertension. AB - The specific competitive alpha 1-postsynaptic blocking action and haemodynamic effects of prazosin (Minipress) have been summarized. Prazosin causes dilatation of arterioles and veins, reduces total peripheral resistance as well as preload and afterload. Cardiac output does not change at rest, stroke volume and subsequent cardiac output increase during exercise. The changes in heart rate have non-significant. It does not cause sympathetic counter-regulation, plasma renin activity does not increase, aldosterone level decreases, salt- and fluid retention may rarely be observed. It does not provoke angina. The authors report on the results of their examinations with the first dose of prazosin in 61 patients (in 33 cases by the double-blind cross-over method by placebo control), and summarize the observations made with the drug in long-term treatment in Hungary. The authors and other teams used prazosin as a long-term treatment (of approximately 3 months) in combination with other drugs in a total of 344 patients, and as monotherapy in 159 patients. In the course of combination treatment side-effects were observed in 15% of the patients (dizziness, headache, weakness, occasionally palpitation). During monotherapy, side-effects occurred in 12% of the patients (tachycardia, headache, weakness, dizziness). Hungarian results confirm the usefulness of prazosin in all stages of hypertension. It is effective in 30-35% of the cases as a monotherapy (this rate is congruent with the efficacy of beta-blockers, calcium antagonists and antihypertensive drugs of central action). Earlier prazosin had been used as a third agent in combination treatment of hypertension.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2573965 TI - Microtubules, organelle transport, and steroidogenesis in cultured adrenocortical tumor cells. 1. An ultrastructural analysis of cells in which basal and ACTH induced steroidogenesis was inhibited by taxol. AB - In adrenocortical cells, the first step in the enzymatic processing of cholesterol to steroid end products occurs in the mitochondria. ACTH increases mitochondrial cholesterol and steroidogenesis. In cultured mouse adrenocortical tumor cells, microtubule-based organelle motility may increase the proximity of mitochondria to the SER, lipid droplets and endoscome-derived lysosomes, thereby facilitating the transfer of cholesterol from these organelles to the mitochondrial outer membrane. ACTH may increase opportunities for the transfer by promoting organelle motility and by increasing the number of lysosomes. Taxol, a microtubule polymerizer, inhibits basal and ACTH-induced steroidogenesis in these cells, presumably at the step where mitochondria obtain cholesterol. We examined the ultrastructure of taxol-treated, unstimulated and ACTH-stimulated cells, seeking alterations which conceivably could interefer with the proposed organelle transport and encounters, and thus correlate with taxol's inhibition of steroidogenesis. Primary cultured cells were incubated in serum-containing medium for 4 hr with and without ACTH (10 mU/ml), with 10 micrograms/ml and 50 micrograms/ml of taxol, and with ACTH and taxol 10 or taxol 50 simultaneously. Culture media were analyzed for the presence of secreted steroids at the end of 1, 2, and 4 hr of incubation. At the end of the fourth hour, unstimulated cells and cells treated with ACTH, taxol 50, and both agents simultaneously, were fixed and processed for EM. Taxol inhibited basal and ACTH-induced steroidogenesis in a dose-dependent fashion. In both unstimulated and ACTH-stimulated cells, taxol 50 formed numerous microtubule bundles, but did not markedly change the distribution of mitochondria and lipid droplets. SER tubules, and clusters of Golgi fragments, endosomes, and lysosomes appeared to be translocated towards the cell periphery along some of the microtubules. Taxol permitted an ACTH-induced cell rounding and microfilament rearrangement considered to facilitate organelle motility. Our data indicate that taxol disrupts the formation of lysosomes by these adrenal cells, but it seemed unlikely that taxol's ultrastructural effects could prevent organelle transport proposed to cause meetings between mitochondria and the SER or lipid droplets, or prevent ACTH-caused increases in these encounters. Taxol may instead prevent the transfer of lipid droplet or SER-contained cholesterol to adjacent mitochondria, by a means not detectable in our electron micrographs. PMID- 2573966 TI - Is there a third heart beta-adrenoceptor? AB - A series of partial agonists with high affinity for myocardial beta 1- and beta 2 adrenoceptors cause stimulant effects in heart that are resistant to blockade of beta 1- and beta 2-adrenoceptors. The concentrations of partial agonist that cause stimulant effects greatly exceed those that cause blockade. Alberto Kaumann suggests that such non-conventional partial agonists, often analogues of pindolol, may act through a third heart beta-adrenoceptor, which resembles the beta 3-adrenoceptor of white adipocytes and smooth muscle of airways and ileum. PMID- 2573967 TI - [The relation of changes in the surface markers and the induction of thymocyte functional activity under the action of humoral thymic factors]. AB - Mouse thymocytes were fractionated on PNA+- and PNA-cells. The PNA+-fraction was treated by tactivin--a preparation of thymic hormones. The treatment resulted in the loss of PNA-receptors from the surface of 10-12% of PNA+-cells. The treated suspension was again fractionated by the same method. PNA-thymocytes derived from both the first and the second procedure of fractionation expressed less Thy-1.2 antigen and SBA-receptors than PNA+-cells. Primary and induced PNA-thymocytes were able to produce IL-2 after mitogen stimulation. The data can be interpreted as an evidence of the ability of thymic hormones to induce not only a phenotypical but also a functional maturation of PNA+-thymocytes and of linkage of all the observed signs of maturation. PMID- 2573968 TI - [Patient information might reduce the use of benzodiazepines in general practice]. AB - Information about benzodiazepines was given during a period of three months in a general practice with two doctors and 2,500 patients in group 1 of the Danish Health Insurance (low income group). The number of requests for repeated prescriptions and the number of defined daily doses of benzodiazepines were registered during periods of one month before and three months after the three month period of information with the object of assessing the patients own wishes for repeated prescription. The investigation revealed an average decrease of 35.4% of requests for repeated prescriptions and a decrease of 55.8% in the defined daily doses prescribed as compared with the month before the information period. The social consequences are discussed. PMID- 2573969 TI - [Sonographic scanning of the non-descended testes]. AB - More active treatment of boys with non-descended testes increases the demand for methods to identify and localize the retained testes. Sonography is non-invasive and is now in common use, but experience with the method is still limited. A retrospective series of 112 examinations, performed in 88 boys, aged 2-16 years, is presented. The indications were uncertainly palpable or non-palpable testes. The results of sonography could be compared with the findings at surgery in 62 instances, 17 testes subsequently descended or were controlled, while 33 examinations were without further verification. Fiftythree testicles were identified at operation, and 50 demonstrated by sonography with a good correlation according to anatomic localization. The suprafascial, everted position was not specified. Abdominal retention was, contrary to previous reports, diagnosed in 86%, when present. Surgery revealed aplasia in nine instances, corresponding to the results of sonography in eight. At a pre operative control-sonography one false positive examination was corrected, and false sonographic diagnoses of aplasia at the first examination in eight instances were reduced to three at preoperative sonographic control. In the group of subsequently descended testicles the result of sonography was considered to be consistent with this cause. It is concluded, that sonography, performed on the indications of uncertainly palpable or non-palpable testes, is of value, since demonstration of a testicle is an accurate diagnosis, and furthermore a correct anatomical localization including abdominal lodging will most often be possible. A single negative ultrasound examination, especially in small boys, should be controlled. PMID- 2573970 TI - [Improvement of primary and secondary prevention of occupational cochlear neuritis]. AB - The methods of primary and secondary prophylaxis of cochlear neuritis in workers exposed to a high noise level are described. It is emphasized that hearing impairment can be prevented in a more efficient manner at the early stages: noise effect on the hearing organ and signs of the effect or mild symptoms of cochlear neuritis. A complete scheme of therapy is presented which includes vasodilators, vitamins, biostimulants, sedatives, physiotherapy. It is stressed that methods of primary and secondary prophylaxis applied in due time can prevent appearance and development of hearing impairment and potential invalidism of workers working in a high noise environment.